PMID- 12882658 TI - CD4+ and CD8+ T cell receptor repertoire perturbations with normal levels of T cell receptor excision circles in HIV-infected, therapy-naive adolescents. AB - Our objective was to determine whether treatment-naive HIV-infected adolescents manifest abnormalities in thymus function and peripheral T cell repertoire, and to assess relationships of these immunologic characteristics with each other, with plasma HIV virus load, and T cell surface markers. TCR Vbeta repertoire was determined by CDR3 length spectratyping in purified CD4(+) and CD8(+) T cells of high-risk, HIV-negative adolescents and of treatment-naive, HIV-infected adolescents. Thymus function was investigated by the simultaneous examination of T cell receptor excision circles (TRECs) in the CD4(+) and CD8(+) T cell subsets. HIV-infected adolescents exhibited significantly greater perturbations in their TCR Vbeta repertoire in comparison with HIV-negative subjects. Perturbations in the CD8(+) T cell compartment were more profound in comparison with CD4(+) T cells. The CD4(+) TCR Vbeta perturbations were negatively correlated with the total and phenotypically naive CD4(+) T cells, and with CD4(+) TRECs. CD8(+) TRECs, although not correlated with CD8(+) TCR Vbeta perturbations, showed negative correlation with memory and activated CD8(+) T cells. Interestingly, TRECs in CD4(+) and CD8(+) T cells were not significantly different between HIV infected and uninfected adolescents. The TCR Vbeta repertoire in adolescents is profoundly perturbed even in early stages of HIV infection, when total CD4(+) cell counts in most subjects are within normal limits. The correlative analyses demonstrating negative association of CD4(+) cell TRECs with CD4(+) TCR Vbeta perturbations and of CD8(+) TRECs with CD8(+) cell activation markers provide evidence of the intense activation of the central and peripheral immune compartments in this study population. PMID- 12882659 TI - Production of CD8+ T cell nonlytic suppressive factors by CD28, CD38, and HLA-DR subpopulations. AB - HIV infection may be modified by CD8(+) T cells by the production of nonlytic antiviral factors. To determine subpopulations that mediate nonlytic, antiviral activity, we examined the production of beta chemokines and of CD8 antiviral factor (CAF) by different subsets, using CD8(+) cells derived from 24 HIV-1 infected and 25 uninfected individuals. Subjects with CD8(+) cell counts greater than 200/microl produced increased levels of MIP-1alpha by CD8(+)CD28(+), CD8(+)CD38(-), and CD8(+)HLA-DR(+) subsets as compared with uninfected controls. CD8(+)CD38(-) cells produced higher levels of MIP-1beta and RANTES. CAF production was increased by CD8(+)CD38(+) and CD8(+)HLA-DR(+) cells of HIV infected individuals as compared with uninfected controls. Chemokine production was increased by cells that do not express activation markers, whereas CAF activity was increased by cells expressing CD38 or HLA-DR. These findings shed light on CD8(+) T cell noncytotoxic antiviral factor production during HIV infection. PMID- 12882660 TI - Identification and characterization of HLA-A*3303-restricted, HIV type 1 Pol- and Gag-derived cytotoxic T cell epitopes. AB - HLA-A*3303 is one of the common HLA alleles in East and Southeast Asia. Identification of HLA-A*3303-restricted HIV-1 cytotoxic T lymphocyte (CTL) epitopes is therefore required to investigate the immunopathogenesis of AIDS and vaccine development in these areas, where AIDS is rapidly expanding. We attempted to identify HLA-A*3303-restricted CTL epitopes derived from relatively conserved proteins Pol, Gag, and Nef of HIV-1 clade B, using reverse immunogenetics. Ninety nine 8-mer to 11-mer peptides corresponding to the HLA-A*3303-binding peptide motif were selected from the HIV-1 SF2 sequence. Fifty-two of these 99 peptides bound to HLA-A*3303. Six of these binding peptides induced peptide-specific CTLs in PBMCs from at least one of two HIV-1-seropositive individuals. CTL clones specific for three Pol peptides and one Gag peptide killed HLA-A*3303-restricted target cells infected with HIV-1 recombinant vaccinia, indicating that these peptides were naturally processed HLA-A*3303-restricted CTL epitopes. SF2-Pol 594 602 (FYVDGAANR) and SF2-Gag 144-152 (MVHQAISPR) induced specific CTLs in 5 and 4 of 10 chronically HIV-1-infected individuals, respectively, whereas SF2-Pol 60-70 (TLWQRPLVTIR) and SF2-Pol 934-943 (KIQNFRVYYR) induced specific CTLs in 2 and 1 of 10 chronically HIV-1-infected individuals, respectively. Thus, the former are immunodominant epitopes whereas the latter are not. These epitopes are useful for studies of AIDS immunopathogenesis and vaccine development. PMID- 12882661 TI - CCR5 delta32 heterozygosity is associated with an increase in CXCR4 cell surface expression. PMID- 12882662 TI - [Long-term results of mitral-aortic valve replacement in 1,154 patients with rheumatic valvular disease]. AB - OBJECTIVE: To analyze the early and long-term results after mitral-aortic valve replacement for rheumatic valvular disease and the determinant factors involved and subsequent therapies. METHODS: 1 154 patients receiving combined mitral aortic valve replacement for rheumatic valvular disease from May 1981 to May 2001 were reviewed. The mean age of the patients was 41.48 +/- 10.00 years. Concomitant valve plasty was performed for associated tricuspid organic or significant functional lesions. Lateral tilting disc or bileaflet valve prostheses were used for replacement. New York Heart Association functional status showed Class III or IV in 91.77% of the patients. Moderate to severe pulmonary hypertension occurred in 29.38% of the patients. The duration of follow up varied from 8 months to 20 years. RESULTS: The hospital mortality was decreased from 6.50% to 4.45%. The 5-, 10- and l5-year survival rates were 89.46% +/- 1.35%, 86.50% +/- l.91% and 67.86% +/- 6.16%, respectively. The 5-, 10- and l5-year thromboembolic event free rates were 97.80% +/- 0.74%, 88.31% +/- 2.20% and 94.08% +/- 2.29%, respectively. the 5-, 10- and l5-year anticoagulant related bleeding free rates were 94.80% +/- 1.09%, 89.32% +/- 2.10% and 83.12% +/- 3.57% respectively. Cardiac functional status returned to Class II in 98% patients and to Class III in 2% during follow-up. CONCLUSIONS: Both left and right ventricular functions may be impaired as a result of rheumatic valvular disease. Tricuspid valve should be explored during surgery and any significant tricuspid annular enlargement and regurgitation showed be corrected in concomitance. Long-acting penicillin regimen is needed for 3 - 5 years for the prevention of rheumatic fever relapse. A low intensity anticoagulant regimen after valve replacement with prothrombin time targeting at 1.5 - 2.0 times is advisable in lessening anticoagulant related bleeding yet optimizing sufficient prevention against thromboembolic complications. PMID- 12882663 TI - [Long-term follow-up of enlargement annulus aortic valve replacement in small aortic root]. AB - OBJECTIVE: To investigate the long-term survival of patients after aortic valve replacement with enlarged annulus. METHODS: From July 1988 to December 2001, the annulus was enlarged in 45 patients. The enlargement techniques included Manouguian's (39 patients), Nicks's (5) and konno's (1). Doppler echocardiography was performed in 43 patients one month after operation. Left ventricular outflow gradient was derived from continuous Doppler measurements of flow velocity, and effective orifice area was calculated by the continuity equation. RESULTS: The operative mortality was 4.4% (2/45) in this group. One patient died of ventricular fibrillation and the other, lower output syndrome. All patients were followed up with a cumulative follow-up period of 6.2 years. Neither valve related deaths nor valve-related complications occurred. 97.6% survivors (42/43) belonged to NYHA class I-II, and 2.4% (1/43), class III. No significant differences were observed in hemodynamic performance of differently sized prostheses for each valve type. CONCLUSIONS: Patient-prosthesis mismatch with heart valve prostheses as demonstrated by the indexed effective orifice area can be avoided by use of Manouguian's annulus enlarging techniques. The hemodynamic performance of these aortic valve prostheses is satisfactory. PMID- 12882664 TI - [Low intensity anticoagulation therapy after mechanical heart valve replacement]. AB - OBJECTIVE: To investigate the proper anticoagulation intensity in patients after mechanical heart valve replacement in china. METHODS: The anticoagulation therapy intensity and the complications in 480 patients after mechanical heart valve replacement were studied. RESULTS: The follow-up rate was 89.17%, the total patient-years (Pty) was 2,110.04 years, the mean oral import warfarin dosage was (2.81 +/- 0.95) mg/day, and native warfarin dosage (2.38 +/- 0.46) mg/day. The mean PTR value of 2 116 samples was 1.43 +/- 0.26, and the INR value of 1 195 samples was 1.63 +/- 0.49. The total hemorrhage rate was 4.60% Pty, and the hemorrhage death rate was 0.38% Pty. The PTR and INR values were higher in the hemorrhage group than in the no-hemorrhage group. (t = 1.816, P < 0.05; t = 2.407, P < 0.01). The thromboembolism rate was 0.66% Pty, and the thromboembolism death rate was 0.05% Pty. There were 15 pregnancies in 14 women patients and no malformed newborns were found. CONCLUSIONS: The most important complication of anticoagulation therapy after mechanical heart valve replacement is hemorrhage in china; The proper anticoagulation intensities of this group are INR 1.5-2.0 and PTR 1.3-1.5. It is beneficial to adopt the low intensity anticoagulation therapy for decreasing the death rate from hemorrhage, protecting pregnant women and newborns from hemorrhage and malformation, and improving the life qualities of the patients. PMID- 12882665 TI - [Long-term results of prosthetic mitral valve replacement with home-made tilting disc valve: a report of 125 cases]. AB - OBJECTIVE: To analyze the early and long-term results after mitral valve replacement for rheumatic valvular disease by using home-made tilting disc valve, and the determinant factors involved and subsequent therapies. METHODS: One hundred and five patients, including 31 patients with rheumatic mitral stenosis, 92 patients with mixed mitral stenosis and regurgitation, and 2 patients with bacterial endocarditis, underwent prosthetic mitral valve replacement with home made tilting disc valve from September 1978 to June 1982. Three patients had a history of mitral commissurotomy, and 5 patients had concomitant functional tricuspid regurgitation. All patients were operated on under cardiopulmonary bypass with implantation of 25 - 29 mm size home-made tilting disc valve prosthesis. The associated functional tricuspid lesions were treated at the same time with modified DeVega's valvuloplasty or Kays bicuspidate valvuloplasty. RESULTS: Eleven patients died during the hospital stay with an early operative mortality of 8.8%. The major causes of the early death were low cardiac output syndrome (4 patients), respiratory failure (2), acute renal failure (2), extrinsic prosthesis dysfunction (1), ventricular arrhythmia (1), and left ventricular rupture (1). Ninety-eight survivors were followed up (total 1,162.2 years) for mean duration of 12.8 years. Eighty-nine patients (78%) survived over 10 years after operation, 58 (51%) over 15 years, and 55 (48%) over 20 years. There were 16 late deaths due to heart failure, anticoagulation related bleeding, thromboembolism and recurrence of rheumatic fever. The survival rates at 10 and 20 years were 82.3% and 51.1% respectively. Among the patients who survived over 20 years, 37 patients had the cardiac functional status returned to Class II, 13 Class III, and Class IV. CONCLUSIONS: Severe post-rheumatic valve deformity may occur in younger patients in China. Long-acting penicillin regimen given for 3 - 5 years for the prevention of rheumatic fever relapse is advocated. A low intensity anticoagulant regimen after mitral valve replacement is advisable in lowering the incidence of anticoagulant related bleeding, while optimizing sufficient protection against thromboembolic complication. Proper operative timing (e.g. when the patient is in sinus rhythm and in NYHA functional class II) is of great importance in achieving satisfied long-term results. PMID- 12882666 TI - [Myocardial revascularization combined with concomitant valve operations: report of 45 cases]. AB - OBJECTIVE: To introduce the experience in coronary artery bypass grafting (CABG) combined with concomitant valve operation. METHODS: From 1998 to 2002, forty-five patients (38 men and 7 women) with coronary artery lesion and valve diseases underwent myocardial revascularization combined with concomitant valve operation. The mean age of the patients was 59 years (42 to 75 years). Heart function was Class II (NYHA) in 7 patients, Class III in 30, Class IV in 8. The value of ejection fraction was 0.32 to 0.50. Thirty patients had mitral valve lesion, 7 aortic lesion, and 8 both lesions. Seventeen patients had single-vessel diseases, 20 double-vessel diseases, 8 triple -vessel diseases, and 8 left main artery lesion. After cardiac arrest by cold blood cardioplegia under moderate cardiopulmonary bypass (CPB), distal anastomosis of the saphenous vein (SV) to the target vessels were firstly performed and followed by valve replacement (32 patients) or plasty (2). The left internal mammary artery (LIMA) was grafted to the left anterior descending artery (LAD) before aortic unclamping. Proximal anastomosis of the SV to the aorta was finally finished on beating heart. The mean bypass time was 173.5 min and the mean duration of aortic cross-clamping time was 112.6 min. RESULTS: No mortality occurred during hospitalization and all patients were discharged 9.2 days after operation. Cardiac function of all patients was improved to Class I-II and no patient died during follow-up. CONCLUSIONS: CABG combined with valve surgery can be safely performed with good results. PMID- 12882667 TI - [Prevention and management of biliary complications following orthotopic liver transplantation]. AB - OBJECTIVE: To prevent and manage biliary complications after orthotopic liver transplantation (OLT). METHODS: Ninety-five patients of OLT performed at our institute from February, 1999 to December 2002 were retrospectively analysed. Recipient operation was performed using standard method combined with veno-venous bypass in 12 patients and piggyback method in 78 patients and living-related liver transplantation in 5 patients. Biliary reconstruction was performed by end to-end choledochocholedochostomy (C-C) over a T-tube in 55 patients and without a T-tube in 36 patients while the remaining 4 patients underwent Roux-en-Y choledochojejunostomy (CRY). C-C and CRY were performed by the interrupted or continuous suture with 5 - 0 or 6 - 0 Vicryl or PDS. Routine examination of liver function, Doppler ultrasonography and cholangiography were performed during the follow-up period. RESULTS: Biliary complications occurred in 7 patients (7.3%). Two patients with bile leakage at the anastomotic site developed biliary peritonitis on the seventh and tenth postoperative day and needed reoperation. One patient developed anastomotic biliary stricture one month after the operation and was cured by endoscopic stenting. Two patients developed bile leakage after T tube removal. One of the two patients was treated conservatively and the other underwent a exploratory laparotomy to ligate the T-tube tract and drain the peritoneal cavity. One patient died of biliary vast syndrome five months after OLT and one patient died of biliary tract necrosis secondary to hepatic artery thrombosis on the tenth postoperative day. One - 42-month (mean 11.4 months) follow-up revealed no biliary stricture in 74 patients. No biliary stone and biliary sludge were detected by Doppler ultrasound and/or cholangiography. Serological examinations proved that liver grafts functioned well in these patients. CONCLUSIONS: To prevent biliary complications, it is crucial to protect biliary mucosa and arterial blood supply of the common bile duct while harvesting the graft and to obtain perfect mucosa-to-mucosa apposition of no-tension end-to end anastomosis of the bile duct. Endoscopic dilation and stenting are effective for post-OLT extrahepatic biliary stricture. PMID- 12882668 TI - [Seventy degrees lateral decubital trans-abdominal laparoscopic adrenal surgery]. AB - OBJECTIVE: To introduce a new procedure of laparoscopic adrenal surgery. METHODS: Thirty patients with adrenal disease underwent surgery by a 70 degrees lateral decubital trans-abdominal laparoscopic approach, which wa compared with 30 degrees decubital laparoscopic operation in 26 patients. RESULTS: The procedure is superior to open laparotomy. There was significant difference in average operative time (77.2 min vs. 215.7 min), blood lost volume (27.0 ml vs. 94.5 ml), and postoperative complications (13.3% vs. 46.0%). Postoperative feeding time and hospitalization time were shortened markedly. CONCLUSIONS: This modified technique is safe, straightforward, with a good anatomic view during operation. It could be widely adopted for the treatment of benign adrenal disease, especially pheochromocytoma. PMID- 12882669 TI - [Nanobacteria in serum, bile and gallbladder mucosa of cholecystolithiasis patients]. AB - OBJECTIVE: To find the distribution of nanobacteria in the serum, bile and gallbladder mucosa of cholecystolithiasis patients. METHODS: The infection rate of nanobacteria was identified by ELISA in the serum samples from 338 healthy people and 76 patients with cholecystolithiasis (chi(2) = 0.89, P > 0.05). Nanobacteria were cultured from the bile samples in 57 patients with cholecystolithiasis and 18 non-cholelithiasis patients and identified by immunohistochemical staining and TEM (chi(2) = 29.80, P < 0.05). Forty samples of gallbladder mucosa randomly selected from the 57 cholecystolithiasis patients were identified by immunohistochemical staining and compared with the corresponding bile samples. RESULTS: The infection rate of nanobacteria was 8.0% and 31.6% for the serum samples of the healthy people and cholecystolithiasis patients, respectively. The positive rate of nanobacteria in the bile samples was 61.3% and there was no significant difference in the bile of the cholecystolithiasis patients and the control group (61.4% vs. 61.1%). Fourteen positive patients had infection of nanobacteria in the gallbladder mucosa, submucosa, and calcific field. CONCLUSIONS: The infection rate of nanobacteria was 8% in the serum samples from the healthy people. There are nanobacteria in the serum, bile, and gallbladder mucosa. The infection of the nanobacteria may result in calcification and fibrosis of the gallbladder. PMID- 12882670 TI - [Therapeutic effects of operation in tumor invades adjacent structures cancer of cardia and stomach fundus]. AB - OBJECTIVE: To study the best style of operation in the treatment of tumor invades adjacent structures (T(4)) cancer of the cardia and stomach fundus. METHODS: Two hundred and one patients with T(4) cancer of the cardia and stomach fundus underwent operation. Of them, 31 were treated by laparotomy, and 170 by combined resection of the involved organs. The 3- and 5-year survival rates and the postoperative complication rate and mortality rate were analyzed in the patients who had under gone combined resection of the involved organs. RESULTS: The median survival of the patients undergoing combined resection of the involved organs (29.3 months) was significantly longer than that of those receiving laparotomy (4.9 months). The 3- and 5-year survival rates of 170 patients who had under gone combined resection of the involved organs were 46.2% and 22.8%, respectively. The 3- and 5-year survival rates of patients undergoing total gastrectomy and proximal gastrectomy were 54.9% and 29.2% and 32.2% and 12.5%, respectively, and the difference was statistically significant (chi(2) = 7.589, P < 0.01;chi(2) = 5.792, P < 0.05).The postoperative mortality rate and complication rate were 4.1% and 24.1%, respectively. CONCLUSIONS: The patients without liver metastasis, widespread nodal involvement, peritoneal dissemination and local focus allowed by an en bloc combined resection in T(4) cancer of cardia and stomach fundus should undergo gastrectomy with a combined resection of the involved organs. Total gastrectomy should be performed to improve the curative effect. PMID- 12882671 TI - [Prospective randomized trial of prophylaxis of postoperative peritoneal carcinomatosis of advanced gastric cancer: intraperitoneal chemotherapy with mitomycin C bound to activated carbon particles]. AB - OBJECTIVE: To evaluate the beneficial effect of intraperitoneally applied mitomycin bound to activated carbon particles (MMC-CH) in the prevention and treatment of intraabdominal recurrence after curative surgery for gastric cancer. METHODS: One hundred and twenty-four patients with radically resected gastric cancer infiltrating the serosal surface were randomly divided into group receiving 50 mg mitomycin bound to a solution of 375 mg carbon adsorbent intraperitoneally before closure of the abdominal wound (n = 62) and a control group (n = 62). The patients with MMC-CH and the control group were received systemic chemotherapy 3 months or 3 weeks after operation respectively. The postoperative recurrence-free survival was evaluated to analyze the benefits of this treatment. RESULTS: After observation for 8 months (range, 2 - 65). The 3-, 5-year postoperative recurrence-free survival rates were significantly higher in the MMC-CH group (70.16%, 44.51%) than in the control group (27.09%, 14.45%), P < 0.01. CONCLUSION: Adjuvant intraperitoneal chemotherapy of gastric cancer by mitomycin bound to activated carbon particles is effected by an increased postoperative recurrence-free survival rate. PMID- 12882672 TI - [Local recurrence and distant metastasis after breast-conserving therapy for patients with breast cancer]. AB - OBJECTIVE: To study the relative factors of local recurrence and distant metastasis after breast-conserving therapy (BCT) for patients with breast cancer. METHODS: The data on 174 patients with primary breast cancer who had been treated by BCT were analyzed retrospectively. The patients were followed up for 12 to 196 months with a rate of 97.13% (169/174). RESULTS: Nine patients showed local recurrence and 14 patients, distant metastasis. The 3-year recurrence rate was 3.79% (5/132), the the 5-year metastasis rate was 10.99% (10/91), and the 5-year survival rate was 92.31% (84/91). Those 3-year recurrence rate for patients without radiotherapy (12.12%) was significantly higher than that with radiotherapy (1.01%), (chi(2) = 5.61, P < 0.05). In patients with node-positive, the the 5-year metastasis rate for patients without chemotherapy (44.44%) was higher than that for those with chemotherapy (6.67%), (P < 0.05). Positive marginal status was associated with local recurrence (P < 0.01). Age at diagnosis of patient with breast cancer ( 0.05); (2) The number of apoptotic cells in MDS group was much higher than that in the controls (71.5 +/- 70.9/mm(2) vs 37.3 +/- 23.0/mm(2), P < 0.05); (3) The number of CD68 expression (35.5 +/- 37.0/mm(2)) and apoptosis (90.7 +/- 74.6/mm(2)) in less advanced MDS were much higher than that in advanced MDS group (14.6 +/- 11.7/mm(2) and 26.8 +/- 33.1/mm(2), P < 0.05 and < 0.01 respectively); (4) CD68 expression showed an obvious positive correlation to apoptosis in MDS cases (r = 0.83, P < 0.001); (5) CD68 positive cells did not show location correlation to apoptotic cells; (6) CD 68 positive cells in MDS showed simultaneous apoptosis. CONCLUSIONS: Over-apoptosis existed in MDS. Less advanced group has a higher ratio of apoptosis than in advanced group. The correlation between macrophages and apoptosis indicates the participation of TNFalpha in apoptosis-induction during MDS development. PMID- 12882688 TI - [Relationship between TIMP-3 expression and promoter methylation of TIMP-3 gene in hepatocellular carcinoma]. AB - OBJECTIVE: To investigate further the possible mechanism of carcinogenesis and portal invasion of hepatocellular carcinoma (HCC). METHODS: Samples of the primary tumors, cancer cells emboli in the portal veins and normal liver tissues adjacent to the tumor were collected from 20 cases of primary HCC. Expression of TIMP-3 (tissue inhibitor of metalloproteinases-3) protein was detected using Western blot. Expression of TIMP-3 mRNA was detected by RT-PCR. Methylation of TIMP-3 gene promoter was detected using methylation-specific PCR (MSP). RESULTS: Expression of TIMP-3 protein and mRNA were obtained in all of the normal liver tissues adjacent to tumor. However, loss of TIMP-3 protein expression was found in 5 and 36 cases respectively in the primary tumors and tumor cell emboli in portal veins. Expression of TIMP-3 protein and mRNA in primary tumors and tumor emboli were significantly lower than that in the normal liver tissues. Promoter methylation of TIMP-3 gene could be detected in primary tumors (7 cases) and cancerous emboli (9 cases) in HCC, while no methylation found in normal liver tissues. In all the HCC cases with promoter gene methylation including primary tumors and cancerous emboli in portal veins, 13 cases showed complete loss and 6 cases showed low expression of TIMP-3 protein and mRNA. Promoter methylation of TIMP-3 was noticed not related with the histological grading of HCC. CONCLUSIONS: There is a close relationship between loss or low expressions of TIMP-3 and carcinogenesis and portal invasion of HCC. The loss and low expression of TIMP-3 gene and protein were caused by methylation of the gene promoter. PMID- 12882689 TI - [Primary adenoid cystic carcinoma of the nasopharynx and its relation to Epstein Barr virus infection]. AB - OBJECTIVE: To investigate the clinicopathologic characteristics of primary nasopharyngeal adenoid cystic carcinoma (NPACC) and its relation to Epstein-Barr virus (EBV) infection in Guangzhou where is a high-incidence area of EBV associated nasopharyngeal carcinoma (NPC). METHODS: 17 cases of NPACC with clinical record and biopsy samples were collected in Guangzhou and their clinical manifestations were reviewed. Besides HE, Alcian blue and PAS, LSAB immunohistochemistry was performed for detecting the expression of a variety of epithelial markers, CD21 and EBV encoded LMP1. EBV encoded early RNAs (EBER) was detected by using in-situ hybridization. Nested PCR was applied for studying the presence of EBV W-fragment in tissues. RESULTS: The ratio of male to female was 7:10. The patients' age ranged from 30 to 63 years, and the median age was 46 years. 14 out of 17 tumors showed markedly local aggressive growth, presenting as T3 or T4. However, only 1 patient had metastasis of an ipsilateral cervical lymph node. The majority of neoplastic cells were basal-cell like in shape and with scanty cytoplasm and a deeply stained nucleus. Intercellular hyaline or mucinous substance was always present in between the carcinoma cells. Cribriform structure formed by the neoplastic cells could be found in 16 out of these 17 biopsies. The NPACC always express the wide-spectrum cytokeratin and the epithelium membrane antigen. Only a few or a small number of carcinoma cells showed nuclear EBER signals in 9 cases (9/17). Concurrently, these 9 NPACCs showed a 192 bp W fragment positive band on electrophoresis gel by nested PCR. LMP1 expression had been found in 5 out of the 9 NPACCs (55.6%) accompanying with EBER-positive carcinoma cells. The EBER-positive infiltrating lymphocytes could also be found in the stroma of 3 out of the 9 EBER-stained NPACC slides. All the tumor cells, including the EBER-positive cell of the 17 NPACCs showed no CD21 expression. CONCLUSIONS: The female is predominant over the male in development of NPACC, which often accompanied with a markedly invasive capacity at the nasopharynx and its neighboring sites. Only a small number of tumor cells, nearly a half of the studied cases were infected with EBV. Therefore, it's postulated that there seems no close relation present between NPACC and EBV infection. PMID- 12882690 TI - [Relationship between the expression of beta-cat, cyclin D1 and c-myc and the occurance and biological behavior of pancreatic cancer]. AB - OBJECTIVE: To study the relationship between the abnormal expression of beta catenin (beta-cat) and the high expressions of cyclin D1 and c-myc and the occurance, proliferation, infiltration, metastasis and prognosis of pancreatic cancer, and to provide rational basis for the clinical diagnosis and treatment. METHODS: Immunohistochemical PicTure trade mark was used to examine the expressions of beta-cat, cyclin D1 and c-myc in 47 cases of the cancerous tissue of pancreas, 12 cases of the pancreatic intraepithelial neoplasia and 10 cases of normal tissue of pancreas, respectively. Pancreatic cancer proliferation cell nuclear antigen (PCNA) was also tested as the index of the extent of proliferation of the pancreatic cancer. RESULTS: beta-cat was expressed normally in the 10 cases of the normal pancreatic tissue, while cyclin D1 and c-myc were negative. The expression rates of beta-cat, cyclin D1 and c-myc in the tissues of the pancreatic intraepithelial neoplasia and the pancreatic cancer had no significant difference [6/12 and 68.1% (32/47), 6/12 and 74.5% (35/47), 5/12 and 70.2% (33/47) respectively;P values were all more than 0.05]. The abnormal expression rate of beta-cat was significantly correlated to the metastasis of the pancreatic cancer and the one-year survival rate (both P < 0.05), but had no relation with the size, the extent of differentiation, the activity of proliferation, or infiltration of the pancreatic cancer (both P > 0.05). The expression rate of cyclin D1 was correlated with the proliferation of the pancreatic cancer and the extent of differentiation (both P < 0.05), but not with the size, infiltration, metastasis, or one-year survival rate of the pancreatic cancer (both P > 0.05). The expression rate of c-myc was not correlated with the size, the extent of proliferation, infiltration, metastasis, or one-year survival rate (both P > 0.05), but closely with the proliferation activity of the cancerous tissue of pancreas (P < 0.05). The abnormal expression of beta-cat and the high expressions of cyclin D1 and c-myc had a parallel relationship with the pancreatic intraepithelial neoplasia and pancreatic cancer (both P < 0.05, gamma = 1.000, 0.845, 0.437, 0.452). CONCLUSIONS: The abnormal expression of beta-cat activates cyclin D1 and c-myc, and results in the unchecked proliferation and differentiation, which may play an important role in the genesis of the pancreatic cancer. The abnormal expression of beta-cat is one of the mechanisms for the spread of pancreatic cancer and an index in the molecular biology to determine the metastasis and prognosis of pancreatic cancer. PMID- 12882691 TI - [Mechanism of antisense epidermal growth factor receptor cDNA in growth suppression of glioblastomas cells]. AB - OBJECTIVE: To study the mechanism of antisense epidermal growth factor receptor cDNA in growth suppression of glioblastomas cells. METHODS: Glioblastoma U87MG cells, which over-express epidermal growth factor receptor (EGFR), were transfected with antisense-EGFR constructs. Several clones with stable expression of lower or undetectable levels of EGFR protein were obtained. The effect of antisense-EGFR on cell differentiation was studied using morphological evaluation and western blotting analysis of glial fibrillary acidic protein (GFAP) expression. The effect of antisense-EGFR on cell cycle was studied by flow cytometry and immunohistochemical analysis of p53, Rb, p16 and CDK4 expressions. The effect of antisense-EGFR on telomerase activity was studied by telomeric repeat amplification protocol (TRAP) assay. RESULTS: U87MG cells that were transfected with antisense-EGFR constructs had smaller cell bodies and longer processes, and expressed higher level of GFAP compared with that of the control cells. Flow cytometric analysis showed that the proportion of cells in G(0)/G(1) phases of the cell cycle in the antisense EGFR cDNA transfected clones increased significantly when compared with control cells, whereas the proportion of cells in S phase decreased markedly. In addition, immunohistochemical analysis showed that the expression of wild-type p53 was significantly increased in the antisense EGFR cDNA transfected clones, whereas the expressions of Rb, p16 and CDK4 were not altered. TRAP assay revealed that telomerase activity in the antisense-EGFR clones was significantly decreased. CONCLUSIONS: Antisense-EGFR transfection inhibits U87MG cell growth by inducing cell differentiation and p53 expression, G(1) cell cycle arrest and inhibition of telomerase activity. PMID- 12882692 TI - [Expression and significance of integrin alpha6beta4 in experimental allergic neuritis]. AB - OBJECTIVE: To investigate the expression of the integrin alpha6beta4 in experimental allergic neuritis (EAN) and the relationship of the integrin alpha6beta4 with functional states of Schwann cells (Sc) as well as the injury and repair of the myelin during EAN. METHODS: EAN was induced in Lewis rats and sciatic nerves were resected in 18 EAN and 3 normal rats. The expression of tissue integrin alpha6beta4 was analyzed during the course of EAN induction and in controls by in situ hybridization and semi-quantitative RT-PCR. RESULTS: The detection of integrin alpha6 and beta4 subunit by hybridization in situ demonstrated that expression of alpha6 subunit present no significant changes during the course of EAN, while expression of beta4 declined in the early phase, showing less positive signals than those of the control, and restored its expression in the later or recovery phase. The changes of expression of integrin alpha6 and beta4 in EAN were confirmed by semi-quantitative PT-PCR, using GAPDH as the internal standard. CONCLUSIONS: The degeneration and injury of Sc caused by inflammation affect the expression of integrin, which shows similar changes in Sc during embryogenesis, indicating alpha6beta4 may be a marker of Sc differentiation and at least an important molecule to mark the course of EAN. The expression of alpha6beta4 correlate with the injury and repair of myelin during EAN. PMID- 12882693 TI - [Bcl-2 antisense oligonucleotides suppress HL-60 cell growth in a SCID mouse mode]. AB - OBJECTIVE: To study the effects of bcl-2 antisense phosphorothioate oligonucleotides (ASPO) on suppression of HL-60 cell growth in SCID mice and to investigate the feasibility of purging leukemia cells plus bcl-2 ASPO used in vitro. METHODS: 1 x 10(7) viable HL-60 cells were treated with 10 micro mol/L bcl 2 ASPO seven days before the intraperitoneal (IP) inoculation to the SCID mice, Treatment with sense oligonucleotides (SPO) was similar as for the controls. 35 days after the inoculation, all the SCID mice of both groups were sacrificed and their peripheral blood, bone marrow, liver and spleen were examined using half nested RT-PCR and histopathology for detecting the appearance and distribution of the HL-60 cells treated beforehand with antisense or sense oligonucleotides respectively. RESULTS: ASPO could down regulate the expression of bcl-2 resulting in both inhibition of growth and induction of apoptosis in treated HL-60 cells, which failed to develop leukemia in SCID mice at all. However, SPO treated HL-60 cells still behaved their own ways and proliferated agressively, and developed leukemia at last. CONCLUSION: The bcl-2 ASPO enables to suppress HL-60 cell growth and prevent the development of leukemia in the SCID mice. The purging leukemia cells used are seemed liable in inhibiting the development of leukemia in SCID mouse model. PMID- 12882694 TI - [Hepatocyte transformation and tumor development induced by hepatitis C virus NS3 N-terminal protein]. AB - OBJECTIVE: To study the effect of hepatitis C virus nonstructural protein 3 N terminal protein (HCV NS3-5') on hepatocyte transformation and tumor development. METHODS: QSG7701 cells were transfected with plasmid pRcHCNS3-5' (expressing HCV NS3 N-terminal protein) by lipofectamine and selected in G418. The expression of HCV NS3 gene and protein was determined by PCR and immunohistochemistry respectively. Biological effect of transfected cells was observed through cell proliferation assay, anchor independent growth, and tumor development in nude mice. The expression of HCV NS3 and c-myc protein in the induced tumor was evaluated by immunohistochemistry. RESULTS: HCV NS3 was strongly expressed in QSG7701 cells transfected with plasmid pRcHCNS3-5' and the positive signal was located in cytoplasm. The HCV NS3 expression and c-myc protein in the induced cytoplasm. Cell proliferation assay showed that the population doubling time in the pRcHCNS3-5' transfected cells was much shorter than that in the pRcCMV and non-transfected cells (24 h, 26 h, 28 h respectively). The cloning efficiencies of transfected cells with pRcHCNS3-5', pRcCMV and non-transfected cells were 33.0%, 1.5%, 1.1% respectively (P < 0.01). Tumor developed in nude mice inoculated with pRcHCNS3-5'transfected cells 15 days after the inoculation. HE staining showed hepatocarcinoma character and immunohistochemistry confirmed HCV NS3 and c-myc expression in the tumor tissue. The positive control group also showed tumor development, while no tumor mass obtained in the nude mice inoculated with pRcCMV and non-transfected cells even 40 days after the injection. CONCLUSION: HCV NS3 N-terminal protein showed cell transformation and tumorigenic features. PMID- 12882695 TI - [Stable expression of human basic fibroblast growth factor gene in retinal pigment epithelial cells by gene transfer procedures]. AB - OBJECTIVE: The purpose of the present study was to explore whether gene transferred retinal pigment epithelial cells (RPE) can constantly express human basic fibroblast growth factor gene (hbFGF) and to study the feasibility of using this procedure for the treatment of retinitis pigmentosa. METHODS: Plasmid carrying hbFGF [using green fluorescent protein (GFP) as a reporter gene] was constructed and transferred into the RPE cells by lipofectamine. The positively expressed cell clones were selected with G418 and cultured for 4 weeks. Expression of GFP gene was identified by the fluorescence microscope. Expression of hbFGF in the RPE cells was determined by in situ hybridization and immunohistochemical methods. RESULTS: Reconstruction enzyme digestion demonstrated that the eukaryotic expression pcFG was constructed correctly. Expression of GFP protein in the gene transferred RPE was detected under the fluorescence microscope. Enzyme digestion and agarose gel electrophoresis analysis showed that the gene transferred RPE expressed the hbFGF gene. CONCLUSIONS: RPE cells are able to express hbFGF gene stably by gene transfer procedures. PMID- 12882696 TI - [Vesicular stomatitis virus G-protein retrovector mediated a herpes simplex virus thymidine kinase gene transduction and expression in the human retinal pigment epithelial cells]. AB - OBJECTIVE: The herpes simplex virus-thymidine kinase suicide gene (HSV-TK) was delivered to the human retinal pigment epithelial cell (RPE) by a new vesicular stomatitis virus G protein (VSV-G) retroviral vector. The growth inhibitory effects of ganciclovir on VSV-G/HSV-TK transfected RPE were studied. METHODS: A VSV-G retrovirus-packaging cell line 293GPG was transferred with retroviral vector plasmas bearing HSV-TK (G1NaCTK) or galactosidase (LacZ gene, G1BgSvNa) to produce a 293GPG/TK cell line or a 293GPG/LacZ cell line, respectively. LacZ activity was assessed following transduction of CRL2302 cells. HSV-TK transfected RPE and NIH3T3 cells (at different titer levels) were treated with ganciclovir. The growth inhibitory effects of ganciclovir on various cell lines were studied. RESULTS: The titer of VSV-G/HSV-TK concentration was 1.2 x 10(8) cfu/ml. At a high titer (MOI = 200), the efficiency of LacZ gene transfer in CRL2302 cells was 58%. Ganciclovir inhibited the growth of cells transfected by HSV-TK. The maximum inhibitory rate (45%) was obtained at cells with a high titer (MOI = 200). CONCLUSION: Our results suggested that VSV-G/LacZ and VSV-G/HSV-TK retroviral vectors are highly efficient for in vitro delivery and stable expression of genes in NIH3T3 cells and human RPE cells. The cells modified by HSV-TK gene are very sensitive to ganciclovir and the cell growth can be inhibited by ganciclovir. PMID- 12882697 TI - [Exclusive gene mapping on retinitis pigmentosa with markers on chromosomes 3 in a Chinese kindred]. AB - OBJECTIVE: To study the relationship between the rhodopsin gene on chromosome 3 and autosomal dominant retinitis pigmentosa (ADRP) in a Chinese kindred. METHODS: Sixteen normal persons and 18 RP patients in a ADRP family were recruited. Genome scan method based on fluorescence labeled (using 3 different labels: 6-FAM, HEX, and NED) microsatellite markers with multiplex PCR system was used to identify loci influencing susceptibility to ADRP. Fourteen microsatellites (D3S1297, D3S1263, D3S1266, D3S1289, D3S1300, D3S3681, D3S1271, D3S1292, D3S1569, D3S1279, D3S1614, D3S1262, D3S1580 and D3S1311) on chromosome 3 were used as genetic markers. Linkage analysis (using Genescan3.0, GeneScan Analysis 2.1, Genotyper 2.1 and Designer sofe system) was performed using these markers. RESULTS: The LOD value was 4.8 mU/L. The serum TG levels was positively correlated with thyroid volume and was higher in female subjects than in male. An increased serum TG was found in subjects of aged 50 in the community with deficient iodine intake. CONCLUSION: Serum TG level is affected by gender, amount of iodine intake, serum TSH level and thyroid volume. PMID- 12882713 TI - [Influence of irbesartan on renal hypertrophy and thickening of glomerular basement-membrane in streptozotocin-induced diabetic rats]. AB - OBJECTIVE: To investigate the influence of angiotensin II receptor antagonist irbesartan (Irb) on renal hypertrophy and thickening of glomerular basement membrane (GBM) in streptozotocin (STZ) induced diabetic rats. METHODS: Sprague Dalwley (SD) rats were randomly divided into three groups: normal control (group N, n = 7), diabetic nephropathy (group DN, n = 6) and diabetic nephropathy treated with Irb (group DNI, n = 7). Diabetes was induced by injection of STZ intraperitoneally after rats had received uninephroectomy. Blood glucose (BG), body weight (BW), urinary albumin excretion (Ualb), 24-hour proteinuria (24hUpro) were observed in the rats at week 4, 8, 12 respectively. Creatinine clearance (Ccr), kidney weight (KW), profile of kidney hypertrophy (KW/BW), renal tissue protein contents (RTP), glomerular area (A(G)), glomerular volume (V(G)), and width of GBM were determined at week 12 when the rats were sacrificed. Renal expression of connective tissue growth factor (CTGF) and transforming growth factor-beta(1) (TGF-beta(1)) were determined by immunohistochemistry. RESULTS: There is no significant difference of BG between group DN and group DNI (P > 0.05). Irb treatment significantly prevented the increase of Ualb excretion and 24hUpro and the deterioration of Ccr in diabetic rats (P < 0.05, P < 0.01 respectively). Furthermore, Irb markedly inhibited the increasing of KW, KW/BW, RTP, AG and VG shown in diabetic rats (P < 0.05, P < 0.01 respectively). More interestingly, we firstly demonstrated that Irb significantly prevented the thickening of GBM [N group: (127.50 +/- 22.14) nm, DN group: (280.38 +/- 52.77) nm, DNI group: (144.07 +/- 24.85) nm] and immunostaining for CTGF (P < 0.05 respectively). In addition, the extent of CTGF expression is positively correlated with the glomerular immunostaining for TGF-beta(1) and size of V(G) (P < 0.01). CONCLUSION: This is the first data to demonstrate that Irb exerts early renal protective role in diabetic nephropathy, possibly through inhibition of renal hypertrophy and renal expression of CTGF. PMID- 12882714 TI - [Relation of resting energy expenditure to respiratory mechanics and arterial blood gases in chronic obstructive pulmonary disease patients]. AB - OBJECTIVE: To investigate the relationships of the rest energy expenditure to the respiratory mechanics and arterial blood gases in chronic obstructive pulmonary disease (COPD) patients. METHODS: Twenty six patients with COPD and 21 healthy subjects were involved in the study. The rest energy expenditure (REE), oxygen consumption (VO(2)), carbon dioxide production (VCO(2)) and respiratory quotient (RQ) were measured with indirect energy measurements of canopy test; the arterial blood gases were measured soon after energy measurements; then routine pulmonary function, P(0.1) and P(IMAX) were measured at rest. RESULTS: (1) The REE/day, P(0.1)/P(IMAX) and heart rate (HR) in the COPD group [(1,577.69 +/- 311.31) kcal, 0.068 +/- 0.026 and (83.46 +/- 11.36) BPM, respectively] were significantly higher than those in the healthy control group [(1,388.29 +/- 194.89) kcal, 0.039 +/- 0.014 and (69.71 +/- 5.73) BPM, respectively, P < 0.05 for all]; the forced expiratory volume in one second (FEV(1))% of predicted, partial pressure of oxygen in artery (PaO(2)) and arterial oxygen saturation (SaO(2)) in the COPD group [(50.46 +/- 21.35)%, (77.72 +/- 8.84) mm Hg and (92.54 +/- 2.55)%, respectively] were significantly lower than those in the control group [(92.29 +/ 11.91)%, (92.50 +/- 3.82) mm Hg and (96.29 +/- 1.87)%, respectively, P < 0.01 for all]. (2) In COPD group, the REE/day was positively correlated with body height, body weight, body mass index, P(0.1)/P(IMAX) and HR (r = 0.57, 0.65, 0.62, 0.41 and 0.51, respectively, P < 0.05 for all), negatively correlated with FEV(1)%, PaO(2) and SaO(2) (r = -0.43, -0.47 and -0.32, respectively, P < 0.05 for all); the P(0.1) was negatively correlated with PaO(2) and SaO(2) (r = -0.62 and -0.53, respectively, P < 0.01). CONCLUSION: The rest energy expenditure of COPD patients was significantly higher than that of healthy subjects. This increase in REE was not only attributed to the airway obstruction and the damaged gas exchange, but may be related to the elevated respiratory drive and dysfunction of respiratory muscles as well. PMID- 12882715 TI - Prostatitis: diagnosis and classification. PMID- 12882716 TI - Doxazosin for the symptomatic treatment of benign prostatic hyperplasia. PMID- 12882717 TI - The role of urodynamics in the diagnosis and treatment of benign prostatic hyperplasia. PMID- 12882718 TI - The role of combination therapy for lower urinary tract symptoms secondary to benign prostatic hyperplasia. PMID- 12882719 TI - Urethral stents in benign prostate hyperplasia. PMID- 12882720 TI - Transurethral hot water balloon thermotherapy for benign prostatic hyperplasia. PMID- 12882721 TI - ProstaLund feedback thermotherapy: a review. PMID- 12882722 TI - How do transurethral needle ablation of the prostate and transurethral microwave thermotherapy compare with transurethral prostatectomy? PMID- 12882724 TI - Chronic prostatitis cohort study. PMID- 12882723 TI - Rofecoxib versus placebo in prostatitis. PMID- 12882725 TI - Treatment response to conventional and novel therapies in chronic prostatitis. PMID- 12882726 TI - Chronic pelvic pain syndrome: diagnosis and management. PMID- 12882727 TI - Advanced therapy for prostatitis: minimally invasive and invasive therapies. PMID- 12882729 TI - [Primary hyperhidrosis: prospective study in 338 patients]. AB - BACKGROUND AND OBJECTIVE: The objective of this report is to study the clinical aspects of the patients with primary hyperhidrosis (PH) and the social and occupational distressing condition. PATIENTS AND METHOD: From January 1998 to October 2002, 338 patients with PH completed a preoperative questionnaire to register: age, gender, profession, associated diseases, familiar hyperhidrosis history, sweating location, associated dermatological lesions and social embarrassesment. 179 patients were asked about their general symptomatology. RESULTS: In 86% of the patients PH started during infancy, 71.5% were female (mean age 28.8 years). A few patients had others diseases and 42.5% had some associated dermatological lesions. In 47.9% of the patients there is family history of PH. 96.4% reported palmar hyperhidrosis, 80.7% plantar PH and 71.3% reported axillary PH, being less frequent in others regions of the body. The most frequent clinical founding associated is facial blushing in 60.3%, 52.3% heart palpitations, 48% muscle stress, 31.8% reported trembling of the hands and 30,8% headache. In reference to social embarrassesment, we observe that relations between friends and professional environment are the most problematic situation. CONCLUSIONS: PH is a pathologic condition starting in infancy, family history of PH is frequent and most patients have some associated dermatological lesions. Excessive sweating is especially common in palms but no exclusively of this region as it extends to others regions with the same intensity. It can be associated with symptomatology suggestive of hyperexcitability of the sympathetic activity like facial blushing, trembling or headache, symptoms difficult to consider whether they are cause or consequence. PMID- 12882728 TI - Prostatitis versus pelvic pain syndrome: immunologic studies. PMID- 12882730 TI - [Prevalence of metabolic syndrome in patients with coronary heart disease]. AB - BACKGROUND AND OBJECTIVE: We aimed to know the prevalence of classical cardiovascular risk factors, hyperinsulinism, insulin resistance and metabolic syndrome (MS) in a population with coronary heart disease (CHD) as compared with a population without personal or familial history of cardiovascular risk factors. SUBJECTS AND METHOD: This was an observational and transversal study of cardiovascular risk factors making up the MS in an adult population. 367 subjects aged 35-79 years (268 men and 99 women) were studied; 185 had CHD (141 men and 44 women) and 182 (127 men and 55 women) were included as a control population. 82 individuals of the control group who had no MS components were selected for the calculation of parameters of hyperinsulinism and the HOMA (Homeostasis Model Assessment) index in order to evaluate the insulin resistance. RESULTS: As compared with the control group, patients with CHD showed higher blood pressure (systolic and dyastolic), body mass index (BMI), triglicerydes and apo B100 concentrations, and lower HDL-c and apo A1 values. They also exhibited higher values of glycemia and insulinemia and consequently a higher HOMA index. The values of insulin and HOMA (percentil 75), obtained from the selected group for this purpose, were 12 U/ml and 3.03, respectively. In the CHD population, 38.28% (95% CI, 31.13-45.95) had hyperinsulinism, 47.02% (95% CI, 39.70-54.47) had insulin resistance (HOMA) and 41.08% (95% CI, 33.98-48.55) had MS. CONCLUSIONS: There was a high prevalence of cardiovascular risk factors in the CHD population. Patients with CHD have higher hyperinsulinism and insulin resistance as well as a greater prevalence of MS. Despite treatment, a clear association exists between MS and CHD in both men and women. PMID- 12882731 TI - [Relevant factors in medico-legal prognosis of whiplash injury]. AB - BACKGROUND AND OBJECTIVE: Whiplash injury (WI) is commonly evaluated in medico legal practice. With the aim of knowing the determining factors of WI's medico legal prognosis, a prospective and observational study was carried out. PATIENTS AND METHOD: One hundred and twenty consecutive patients who were clinically observed and evaluated in the Medico-Legal Clinic of Barcelona were studied. Socio-demographic, clinical, radiographic and evolutive factors were analyzed. RESULTS: We included 120 patients with a mean age of 35.6 (14) years (range, 4 74), with a homogeneous male/female distribution. An earlier cervical pathology was detected in 10% of patients; none of them had previous psychiatric pathology. 95% corresponded to road-traffic accident cases and there were 5 aggression cases. Over 50% of cases involved a rear-end collision. All patients had neck pain, almost 25% had headache and 13% had paresthesia. According to the Whiplash Association Disorders clinical classification, distribution in grades (G) was: G I 51%, G II 32% and G III 17%. Patients reported recovery within a mean time of 71.6 (46) days (range, 4-244), with 51,2 (45) no working days (range, 0-180 days). The 52% of the patients rest with complains. According to the recovery time, the following medico-legal prognostic factors were identified: age (p < 0.001), being female (p = 0.001), clinical grade (p < 0.001) and abnormal cervical MRI exploration (p < 0.001). Patients with previous cervical pathology reported more complaints (p = 0.001). CONCLUSIONS: In our study, WI affected young people of both sex mainly during rear-end collision. Worst medico-legal prognostic factors were age, being females, severity of initial clinical symptoms, previous cervical pathology and abnormal cervical MRI/CT. PMID- 12882732 TI - [Familial susceptibility to aminoglycoside ototoxicity due to the A1555G mutation in the mitochondrial DNA]. AB - BACKGROUND AND OBJECTIVE: The A1555G mutation in the mitochondrial genome causes sensorineural hearing loss and familial aminoglycoside ototoxicity. PATIENTS AND METHOD: Screening for the A1555G mutation was performed on 72 patients with nonsyndromic sensorineural hearing loss. RESULTS: The A1555G mutation was identified in 15 patients (20.8%). All of them presented maternal relatives with deafness. Individuals with the A1555G mutation that had been treated with aminoglycosides developed more severe hearing loss. CONCLUSIONS: The A1555G mutation should be screened in individuals with maternal relatives with hearing loss before administering aminoglycosides. PMID- 12882733 TI - [Hypnosis as a treatment of pain]. PMID- 12882734 TI - [Myocardial infarction: fibrinolytic treatment in the emergency room]. PMID- 12882735 TI - [Abridged checklist for the evaluation of basic biomedical research papers]. PMID- 12882736 TI - [Role of ultrasound-guided endoscopy in the evaluation of mediastinal lesions]. AB - The evaluation of the mediastinum is still a challenge. In most cases, a pathologic study is mandatory for therapeutic decision-making. In this setting, endoscopic ultrasonography (EUS) without and with fine needle aspiration (FNA) is currently considered as a very useful and safe non-invasive technique since it overcomes most problems raising from standard radiologic techniques at the time of obtaining histologic confirmation. Moreover, it avoids the surgical approach for diagnosis in a significant number of patients. Thus, performance characteristics (sensitivity, specificity and accuracy) of EUS FNA in the diagnosis of mediastinal lymph nodes of unknown origin are higher than 90%. The place of EUS FNA in the staging of lung cancer is not well established yet, but available data suggest that it can play a major role in patients with negative transbronchial biopsy or even in the initial evaluation regardless of CT results. Finally, recent studies also suggest that EUS FNA is the most cost-effective non surgical technique for the study of the mediastinum. PMID- 12882737 TI - [Intravascular catheter-related infections]. PMID- 12882738 TI - [Ocular dirofilariosis probably caused by Dirofilaria immitis]. PMID- 12882740 TI - [Human metapneumovirus: a new agent in the differential diagnosis of respiratory tract infection]. PMID- 12882741 TI - [Evaluation of two rapid tests for the determination of microalbuminuria and the urinary albumin/creatinine ratio]. AB - OBJECTIVE: Microalbuminuria screening is justified on the grounds of its cost benefit ratio in patients at risk of kidney damage while the process is still reversible. The aim of the present study was to evaluate the DCA 2000 analyser and the Clinitek 50 system (Bayer), which simultaneously measure urinary albumin and creatinine levels to adopt them as rapid methods for microalbuminuria detection. METHODS: One hundred twenty-seven urine samples from pediatric patients with various disorders were assessed. Albumin, creatinine, and the albumin/creatinine ratio were determined using the DCA 2000 analyzer and the Clinitek 50 system, which were compared against the usual reference laboratory methods. RESULTS: The correlation coefficient of nephelometric values vs the DCA 2000 analyzer was 0.914 for albumin, 0.970 for creatinine and 0.839 for the albumin/creatinine ratio. At an albumin cut-off concentration of 30 mg/l, the sensitivity, specificity, positive predictive value and negative predictive value were 100 %, 93 %, 84 % and 100 % for the DCA 2000 analyzer and 91.7 %, 86 %, 55 % and 98 % for the Clinitek 50 system. ROC curve analysis showed that the DCA 2000 system was more effective than the Clinitek 50 in microalbuminuria screening. CONCLUSIONS: The data obtained with the DCA 2000 system showed close agreement with those obtained with reference laboratory methods. The immediate availability of results is a great advantage in clinical practice. The Clinitek-Microalbumin dipstick system is a semiquantitative method that is easy to use, low in cost, simple and useful for screening, but it is less reliable as a follow-up method. PMID- 12882742 TI - [Use of goat's milk in patients with cow's milk allergy]. AB - BACKGROUND: Several epidemiological studies estimate that cow's milk allergy affects approximately 3-5 % of infants in industrialized countries. Infant formulae based on soy protein or hydrolysates free of cow's milk protein guarantee adequate nutritional development, although they may be inadequate for older patients. OBJECTIVE: To evaluate the possibility of using goat's milk in these patients by investigating possible cross-reactivity between both proteins in vivo and in vitro. METHODS: Twelve patients with cow's milk allergy were tested for tolerance to goat's milk protein through radioallergosorbent assay (RAST), specific IgE, skin prick and challenge tests. RESULTS: Only 25 % of the patients showed adequate immediate and late oral tolerance and had negative results of immunological tests for adverse reactions. CONCLUSIONS: The use of goat's milk cannot be recommended to patients with cow's milk allergy without investigation of possible tolerance by a specialist. For the 25 % of patients that tolerate goat's protein, goat's milk can be a excellent substitute in children older than 2 years old. PMID- 12882743 TI - [Topotecan for pediatric patients with resistant and recurrent solid tumors]. AB - BACKGROUND: Topotecan is a cytotoxic drug isolated from the Camptotheca acuminata tree (from China). It is able to block the enzyme DNA topoisomerase I and has recently been used in the treatment of pediatric cancer. OBJECTIVES: To evaluate our preliminary experience with topotecan in the second line treatment of refractory solid tumors in the pediatric age group. PATIENTS AND MEHTODS: We performed a retrospective study of 10 patients with various recurrent solid tumors resistant to first line treatment who were treated with topotecan alone or in association with other chemotherapeutic agents. RESULTS: Ten patients with recurrent solid tumors or tumors that were refractory to conventional treatment (two neuroblastomas, three rhabdomyosarcoma, two PNET/Ewing's sarcoma, one anaplastic astrocytoma, one soft tissue sarcoma and one synovial sarcoma) were included. Five patients showed favorable responses (two had complete responses, two had partial responses and one had stable disease). Five patients showed no response. All patients showed grade II-IV hematological toxicity. CONCLUSIONS: In our experience, topotecan is beneficial in some refractory or recurrent solid tumors, especially neuroblastomas and soft tissue sarcomas. Myelosuppression was tolerable with the use of granulocyte colony-stimulating factors. Patients with a complete response to topotecan could benefit from high-dose chemotherapy and autologous stem cell rescue therapy. PMID- 12882744 TI - [Serum selenium levels in neonates]. AB - BACKGROUND: Despite their increasing importance in children's nutrition, studies on selenium levels in neonates in Spain are scarce and often contradictory. OBJECTIVES: To establish the standard serum levels of selenium in healthy full term neonates in our area and to contribute knowledge of the perinatal factors that influence these levels. METHODS: We determined selenium levels in serum by atomic absorption spectrophotometry in 247 neonates: 70 healthy full term neonates, 60 sick full term neonates, 18 neonates with intrauterine growth retardation (> 37 weeks; birthweight < 2500 g), 44 healthy preterm neonates and 55 sick preterm neonates. RESULTS: Healthy full term newborns showed higher serum selenium levels than healthy preterm neonates (35.11 6.94 g/l, range: 18.4-48 g/l versus 28.65 5.95 g/l, range: 15-44.4 g/l, p < 0.001). In the group with intrauterine growth retardation, serum selenium levels were higher than in the healthy preterm group (30.80 6.97 g/l, range: 20-45.6 g/l versus 28.65 5.95 g /L, range: 15-44.4 g/l) but lower than in the full term and normal birthweight group (idem versus 35.11 6.94 g/l, range 18.4-48 g/l). Likewise, the low birthweight group (< 2500 g) showed lower mean serum selenium levels than the normal birthweight group (27.98 6.75 g/l, range 15-48 g/l versus 33.09 7.52 g/l, range 14.4-48 g/l; p < 0.001). CONCLUSIONS: Prematurity and low birthweight are the best predictors for risk of neonatal hyposeleniemia. PMID- 12882745 TI - [Mechanical ventilation in pediatrics (III). Weaning, complications and other types of ventilation. Weaning from mechanical ventilation]. AB - Weaning from mechanical ventilation can be defined as the process that allows the transition from mechanical ventilation to spontaneous breathing. This process can account for a significant proportion of total ventilation time and failure to resume spontaneous breathing affects patient outcome. Thus, to ensure maximum success, patient readiness for weaning and extubation should be evaluated through the following steps: the patient must fulfill pre-established clinical and ventilatory support criteria for extubation, the patient should be observed during a breathing trial on minimal or no ventilatory support, and variables used to predict weaning success should indicate a favorable outcome. Breathing trials are usually conducted while the patient breathes spontaneously through a T-tube system or through the ventilator circuit on minimal ventilatory support. Neither of these methods has proved superior to the other. The best prognostic indicator of weaning outcome is clinical assessment of respiratory effort. Once mechanical ventilation is discontinued, it may be necessary to treat post-extubation complications or even to resume ventilatory support. PMID- 12882746 TI - [Subacute paresis of the lower limbs]. PMID- 12882747 TI - [Diagnosis, treatment and long-term outcome in Cushing's disease]. AB - Cushing's disease is uncommon in childhood. It is usually caused by the oversecretion of adrenocorticotrophic hormone (ACTH) by a pituitary corticotroph adenoma. Clinical abnormality is often undetected for long periods of time. The principal clinical features are growth retardation and obesity. We present four patients who were diagnosed with Cushing's disease in a tertiary hospital from January 1995 to December 2002. There were two boys and two girls, aged 10-15 years. The main clinical features at presentation were growth failure and excessive weight gain. The interval between onset of symptoms and diagnosis ranged from 2.5 to 5 years. All patients presented moon face and increased fat around the neck. In all patients, 24-hour urinary free cortisol was high and loss of normal serum cortisol circadian rhythm (3/3) and suppression of cortisol to less than 50 % of the basal level with high-dose but not with low-dose dexamethasone (2/2) were observed. Pituitary microadenoma was detected by magnetic resonance imaging in three patients; no tumor was detected in the fourth patient and inferior petrosal sinus sampling was performed, showing left lateralization of ACTH central secretion. Therapy consisted of transsphenoidal surgery in all patients. Two patients are in remission, one patient presented disease recurrence requiring total hypophysectomy because the tumor could not be completely removed and one patient had persistent disease after a second intervention and required pituitary radiotherapy. Two patients showed pituitary hormone deficiency after therapy. PMID- 12882748 TI - [Rasmussen's syndrome, an autoimmune disease]. AB - Rasmussen's disease is an inflammatory, chronic and progressive brain disorder that usually presents with neocortical focal seizures resistant to conventional treatment and culminates in severe deterioration with hemiparesis, cognitive decline and aphasia. Viral infections and antibodies to the GluR3 receptor have been implicated in the physiopathology of this illness and T-cell mediation may play a role in the cerebral inflammatory process. Classical treatment consists of hemispherectomy of various magnitudes depending on cerebral involvement. The association between therapy-resistant epilepsy and autoimmune phenomena due to antibodies against glutamic acid decarboxylase (anti-GAD) have very recently begun to be studied. The discovery of this association led to a new focus and alternative therapies with immunosuppressors, immunoglobulins, steroids and plasmapheresis, alone or in combination, have begun to be tested with variable success. We describe a boy who was diagnosed in the early stages of Rasmussen's syndrome. He tested positive for anti-GAD antibodies and received treatment with immunoglobulins and steroids. After treatment the boy tested negative for anti GAD antibodies and he remains asymptomatic after ten months. PMID- 12882749 TI - [Chorioretinal dysplasia-microcephaly-mental retardation syndrome]. PMID- 12882750 TI - [Giant ovarian teratoma: an incidental finding]. PMID- 12882751 TI - [Cholestatic hepatitis as initial manifestation of scarlet fever]. PMID- 12882752 TI - [Autoimmune hemolytic anemia due to biphasic hemolysin]. PMID- 12882755 TI - Genome-wide search and identification of a novel gel-forming mucin MUC19/Muc19 in glandular tissues. AB - Gel-forming mucins are major contributors to the viscoelastic properties of mucus secretion. Currently, four gel-forming mucin genes have been identified: MUC2, MUC5AC, MUC5B, and MUC6. All these genes have five major cysteine-rich domains (four von Willebrand factor [vWF] C or D domains and one Cystine-knot [CT] domain) as their distinctive features, in contrast to other non-gel-forming type of mucins. The CT domain is believed to be involved in the initial mucin dimer formation and have very succinct relationship between different gel-forming mucins across different species. Because of gene duplication and evolutional modification, it is very likely that other gel-forming mucin genes exist. To search for new gel-forming mucin candidate genes, a "Hidden Markov Model"(HMM) was built from the common features of the CT domains of those gel-forming mucins. By using this model to screen all protein databases as well as the six-frame translated expression sequence tag and translated human genomic databases, we identified a locus located at the peri-centromere region of human chromosome 12 and the corresponding homologous region of mouse chromosome 15. We cloned the 3' end of this gene and its mouse homolog. We found one vWF C domain, one CT domain, and various mucin-like threonine/serine-rich repeats. Phylogenetic analysis indicated the close relationship between this gene and the submaxillary mucin from porcine and bovine. A polydispersed signal was observed on the Northern blot, which indicates very large mRNA size. Further analysis of the upstream genomic sequences generated from human and mouse genome projects revealed three additional vWF D domains and many mucin-like threonine/serine-rich repeats. The expression of this gene is restricted to the mucous cells of various glandular tissues, including sublingual gland, submandibular gland, and submucosal gland of the trachea. Based on the chronological convention, we have given the name MUC19 to the human ortholog and Muc19 to the mouse. PMID- 12882756 TI - Lung cancer screening: a different paradigm. AB - Thoracic computed tomography (CT) is a sensitive method for detecting early lung cancer but has a high false-positive rate and is not sensitive for detecting central preinvasive and microinvasive cancer. Our hypothesis was that automated quantitative image cytometry (AQC) of sputum cells as the first screening method may improve detection rate by identifying individuals at highest risk for lung cancer. A total of 561 volunteer current or former smokers 50 years of age or older, with a smoking history of more than or equal to 30 pack/years, were studied. Among these, 423 were found to have sputum atypia defined as five cells or more with abnormal DNA content using AQC. Noncalcified pulmonary nodules were found in 46% (259/561). Of the 14 detected cancers, 13 were detected in subjects with sputum atypia-nine by CT and four carcinoma in situ/microinvasive cancers by autofluorescence bronchoscopy. One cancer was detected by CT alone. AQC of sputum cells improved the detection rate of lung cancer from 1.8 to 3.1%. CT scan alone would have missed 29% of the cancers. This screening paradigm shift has the additional potential of reducing the number of initial CT scans by at least 25% with further savings in follow-up investigations and treatment. PMID- 12882757 TI - C-C chemokine receptor 2 and sarcoidosis: association with Lofgren's syndrome. AB - Sarcoidosis is thought to result from the interaction between an unknown environmental antigenic trigger and the host's genetic susceptibility. We hypothesized that sarcoidosis, or one of the disease subsets, could be associated with single nucleotide polymorphisms of C-C chemokine receptor 2 (CCR2) gene. Eight single-nucleotide polymorphisms in CCR2 were studied in a total of 304 Dutch individuals (90 non-Lofgren sarcoidosis, 47 Lofgren's syndrome, 167 control subjects). From the investigated CCR2 polymorphisms, nine haplotypes were deduced (haplotypes 1-9). In patients with Lofgren's syndrome, a strongly significant increase in the frequency of CCR2-haplotype 2, which includes four unique alleles (A at nucleotide position -6752, A at 3,000, T at 3,547, and T at 4,385), was observed compared with control subjects (74% vs. 38% respectively, p < 0.0001), whereas no difference was found between non-Lofgren sarcoidosis and control subjects (both 38%). The association between CCR2-haplotype 2 carriage frequency and Lofgren's syndrome (odds ratio, 4.4; p < 0.0001) remained significant after adjustment for human leukocyte antigen haplotype DRB1*0301-DQB1*0201 (odds ratio, 11.5; p < 0.0001) and female sex (odds ratio, 3.2; p = 0.003), two known risk factors for Lofgren's syndrome. In conclusion, this report describes a strong association between CCR2-haplotype 2 and Lofgren's syndrome. Further studies are needed to understand the molecular mechanisms underlying this association. PMID- 12882758 TI - Surfactant protein-A and phosphatidylglycerol suppress type IIA phospholipase A2 synthesis via nuclear factor-kappaB. AB - We previously showed that surfactant inhibits the synthesis of type IIA secretory phospholipase A2 (sPLA2-IIA) by alveolar macrophages. These cells have been identified as the main source of this enzyme in an animal model of acute lung injury. The aim of the present study was to identify the surfactant components involved in the inhibition of sPLA2-IIA expression in alveolar macrophages and the signaling pathways that mediate this inhibition. Our results show that various surfactant preparations can inhibit sPLA2-IIA expression in endotoxin stimulated alveolar macrophages. Both the surfactant protein (SP)-A and the surfactant phospholipid fraction inhibit this expression. The surfactant phospholipid dioleylphosphatidylglycerol (DOPG) abolishes sPLA2-IIA expression, whereas dipalmitoylphosphatidylcholine does not. Chromatographic analysis and confocal microscopy revealed that phosphatidylglycerol was rapidly incorporated and metabolized by alveolar macrophages and that its metabolites accumulate in the cytosol. Nuclear factor-kappaB (NF-kappaB) modulates sPLA2-IIA expression in endotoxin-activated alveolar macrophages, and surfactant preparations, surfactant phospholipid fraction, SP-A, and DOPG indeed suppressed NF-kappaB activation. In summary, our results show that SP-A and DOPG play a role in the surfactant mediated inhibition of sPLA2-IIA expression in alveolar macrophages and that this inhibition occurs via a downregulation of NF-kappaB activation. PMID- 12882759 TI - Surfactant protein D regulates airway function and allergic inflammation through modulation of macrophage function. AB - The lung collectin surfactant protein D (SP-D) is an important component of the innate immune response but is also believed to play a role in other regulatory aspects of immune and inflammatory responses within the lung. The role of SP-D in the development of allergen-induced airway inflammation and hyperresponsiveness (AHR) is not well defined. SP-D levels progressively increased up to 48 hours after allergen challenge of sensitized mice and then subsequently decreased. The levels of SP-D paralleled the development of airway eosinophilia and AHR. To determine if this association was functionally relevant, mice were administered rat SP-D (rSP-D) intratracheally. When given to sensitized mice before challenge, AHR and eosinophilia were reduced by rSP-D in a dose-dependent manner but not by mutant rSP-D. rSP-D administration resulted in increased levels of interleukin (IL)-10, IL-12, and IFN-gamma in bronchoalveolar lavage fluid and reduced goblet cell hyperplasia. Culture of alveolar macrophages together with SP-D and allergen resulted in increased production of IL-10, IL-12, and IFN-gamma. These results indicate that SP-D can (negatively) regulate the development of AHR and airway inflammation after airway challenge of sensitized mice, at least in part, by modulating the function of alveolar macrophages. PMID- 12882760 TI - Pulmonary cytochrome P-450 2J4 is reduced in a rat model of acute Pseudomonas pneumonia. AB - We previously reported that the levels of epoxyeicosatrienoic acids (EETs) and 20 hydroxyeicosatetraenoic acid (20-HETE) are depressed in microsomes prepared from lungs of rats with acute Pseudomonas pneumonia. We also showed a potential role for cytochrome P-450 (CYP) metabolites of arachidonic acid (AA) in contractile responses of both normal pulmonary arteries and pulmonary arteries from rats with pneumonia. The CYP2J subfamily enzymes (endogenous source of EETs and HETEs) are constitutively expressed in human and rat lungs where they are localized in vascular smooth muscle and endothelium. The purpose of this study was to determine if CYP2J proteins are modified in pneumonia. Pseudomonas organisms were injected via a tracheostomy in the lungs of rats. Later (44 h), lungs were frozen, and microsomes were prepared from pneumonia and control rat lung homogenates. Lung microsomal proteins were then immunoblotted with anti CYP2B1/2B2, anti-CYP4A, anti-CYP2J9pep2 (which reacts with rat CYP2J3), anti CYP2J6pep1 (which reacts with rat CYP2J4), anti-CYP2J2pep4, or anti-CYP2J2pep3 (both of which react with all known CYP2J isozymes). Western blotting revealed a prominent 55-kDa band with anti-CYP2J2pep3, anti-CYP2J2pep4, and anti-CYP2J6pep1 (but not anti-CYP2J9pep2) that was reduced in pneumonia compared with control lung microsomes. The CYP2B bands (51-52 kDa) were less prominent and not different between pneumonia and control lungs. CYP4A proteins (20-HETE sources) were not detected in rat lung microsomes. Therefore, rat lung contains a protein with immunological characteristics similar to CYP2J4, and this CYP is reduced after pneumonia. We speculate that CYP2J (but not CYP2B) enzymes and their AA metabolic products (EETs) are involved in the modulation of pulmonary vascular tone in pneumonia in rats. PMID- 12882761 TI - Chronic intrauterine pulmonary hypertension compromises fetal pulmonary artery smooth muscle cell O2 sensing. AB - To test the hypothesis that chronic intrauterine pulmonary hypertension (PHTN) compromises pulmonary artery (PA) smooth muscle cell (SMC) O2 sensing, fluorescence microscopy was used to study the effect of an acute increase in Po2 on the cytosolic Ca2+ concentration ([Ca2+]i) of chronically hypoxic subconfluent monolayers of PA SMC in primary culture. PA SMCs were derived from fetal lambs with PHTN due to intrauterine ligation of the ductus arteriosus. Acute normoxia decreased [Ca2+]i in control but not PHTN PA SMC. In control PA SMC, [Ca2+]i increased after Ca2+-sensitive (KCa) and voltage-sensitive (Kv) K+ channel blockade and decreased after diltiazem treatment. In PHTN PA SMC, KCa blockade had no effect, whereas Kv blockade and diltiazem increased [Ca2+]i. Inhibition of sarcoplasmic reticulum Ca2+ ATPase activity caused a greater increase in [Ca2+]i in controls compared with PHTN PA SMC. Conversely, ryanodine caused a greater increase of [Ca2+]i in PHTN compared with control PA SMC. KCa channel mRNA is decreased and Kv channel mRNA is unchanged in PHTN PA SMC compared with controls. We conclude that PHTN compromises PA SMC O2 sensing, alters intracellular Ca2+ homeostasis, and changes the predominant ion channel that determines basal [Ca2+]i from KCa to Kv. PMID- 12882762 TI - Heparin-binding EGF-like growth factor regulates elastin and FGF-2 expression in pulmonary fibroblasts. AB - Elastase degradation of elastin within alveolar walls is an important event in the development of pulmonary emphysema. In addition to elastolytic activities, elastases release growth factors from extracellular matrices and interstitial cell surfaces that can regulate elastogenesis and other cellular responses. In the present study, we demonstrate that brief treatment of matrix-laden rat pulmonary fibroblast cultures with pancreatic elastase results in the release of soluble heparin-binding epidermal growth factor-like growth factor (HB-EGF) concomitant with a decrease in HB-EGF binding to both heparan sulfate proteoglycan and receptor sites on the cells. In undigested, matrix-laden fibroblasts, HB-EGF significantly downregulates elastin mRNA via activation of epidermal growth factor receptor. Results from nuclear run-on analyses show that HB-EGF downregulates elastin mRNA via transcriptional suppression. HBEGF treatment stimulates MAP or ERK kinase (MEK)-dependent ERK1/2 phosphorylation and leads to nuclear accumulation of Fra-1. Blocking ERK1/2 activation by MEK1/2 inhibitors (PD-98059 or U-0126) diminishes HB-EGF-induced Fra-1 accumulation and subsequent downregulation of elastin mRNA. Coaddition of two elastase-released growth factors, HB-EGF and FGF-2, results in an additive inhibitory effect on elastin mRNA levels. Furthermore, HB-EGF addition to pulmonary fibroblasts increases FGF-2 mRNA and protein levels. These data suggest that HB-EGF and FGF-2 act in concert to regulate the synthesis of elastin in injury/repair situations. PMID- 12882763 TI - Unbalanced collagenases/TIMP-1 expression and epithelial apoptosis in experimental lung fibrosis. AB - In this study, we examined the sequential expression of several matrix metalloproteinases (MMPs), tissue inhibitors of metalloproteinases (TIMPs), and growth factors as well as the presence of apoptosis in a model of pulmonary fibrosis induced in rats with paraquat and hyperoxia. Animals showing neither clinical nor morphological changes with this double aggression were classified as "resistant". Rats were killed at 1, 2, 3, and 6 wk, and lungs were used for collagen content, gene expression by real-time PCR, gelatinolytic activity by zymography, apoptosis by in situ DNA fragmentation, and protein localization by immunohistochemistry. Our results showed a significant decrease of collagenases MMP-8 and MMP-13, with an increase of TIMP-1 and transforming growth factor-beta. Immunoreactive TIMP-1 was increased in experimental rats and primarily localized in alveolar macrophages. Expression of gelatinases MMP-2 and MMP-9 mRNAs was not affected, but lung zymography revealed an increase in progelatinase B, progelatinase A, and its active form. Epithelial apoptosis was evident from the first week, whereas at later periods, interstitial cell apoptosis was also noticed. Resistant animals behave as controls. These findings suggest that an imbalance between collagenases and TIMPs, excessive gelatinolytic activity, and epithelial apoptosis participate in the fibrotic response in this experimental model. PMID- 12882764 TI - Duroquinone reduction during passage through the pulmonary circulation. AB - The lungs can substantially influence the redox status of redox-active plasma constituents. Our objective was to examine aspects of the kinetics and mechanisms that determine pulmonary disposition of redox-active compounds during passage through the pulmonary circulation. Experiments were carried out on rat and mouse lungs with 2,3,5,6-tetramethyl-1,4-benzoquinone [duroquinone (DQ)] as a model amphipathic quinone reductase substrate. We measured DQ and durohydroquinone (DQH2) concentrations in the lung venous effluent after injecting, or while infusing, DQ or DQH2 into the pulmonary arterial inflow. The maximum net rates of DQ reduction to DQH2 in the rat and mouse lungs were approximately 4.9 and 2.5 micromol. min(-1).g dry lung wt(-1), respectively. The net rate was apparently the result of freely permeating access of DQ and DQH2 to tissue sites of redox reactions, dominated by dicumarol-sensitive DQ reduction to DQH2 and cyanide sensitive DQH2 reoxidation back to DQ. The dicumarol sensitivity along with immunodetectable expression of NAD(P)H-quinone oxidoreductase 1 (NQO1) in the rat lung tissue suggest cytoplasmic NQO1 as the dominant site of DQ reduction. The effect of cyanide on DQH2 oxidation suggests that the dominant site of oxidation is complex III of the mitochondrial electron transport chain. If one envisions DQ as a model compound for examining the disposition of amphipathic NQO1 substrates in the lungs, the results are consistent with a role for lung NQO1 in determining the redox status of such compounds in the circulation. For DQ, the effect is conversion of a redox-cycling, oxygen-activating quinone into a stable hydroquinone. PMID- 12882765 TI - Effect of surfactant protein A on granular pneumocyte surfactant secretion in vitro. AB - Surfactant secretion by lung type II cells occurs when lamellar bodies (LBs) fuse with the plasma membrane and surfactant is released into the alveolar lumen. Surfactant protein A (SP-A) blocks secretagogue-stimulated phospholipid (PL) release, even in the presence of surfactant-like lipid. The mechanism of action is not clear. We have shown previously that an antibody to LB membranes (MAb 3C9) can be used to measure LB membrane trafficking. Although the ATP-stimulated secretion of PL was blocked by SP-A, the cell association of iodinated MAb 3C9 was not altered, indicating no effect on LB movement. FM1-43 is a hydrophobic dye used to monitor the formation of fusion pores. After secretagogue exposure, the threefold enhancement of the number of FM1-43 fluorescent LBs (per 100 cells) was not altered by the presence of SP-A. Finally, there was no evidence of a large PL pool retained on the cell surface through interaction with SP-A. Thus SP-A exposure does not affect these stages in the surfactant secretory pathway of type II cells. PMID- 12882766 TI - Role of Ca2+ in diperoxovanadate-induced cytoskeletal remodeling and endothelial cell barrier function. AB - Diperoxovanadate (DPV), a potent inhibitor of protein tyrosine phosphatases and activator of tyrosine kinases, alters endothelial barrier function via signaling pathways that are incompletely understood. One potential pathway is Src kinase mediated tyrosine phosphorylation of proteins such as cortactin that regulate endothelial cell (EC) cytoskeleton assembly. As DPV modulates endothelial cell signaling via protein tyrosine phosphorylation, we determined the role of DPV induced intracellular free calcium concentration ([Ca2+]i) in activation of Src kinase, cytoskeletal remodeling, and barrier function in bovine pulmonary artery endothelial cells (BPAECs). DPV in a dose- and time-dependent fashion increased [Ca2+]i, which was partially blocked by the calcium channel blockers nifedipine and Gd3+. Treatment of cells with thapsigargin released Ca2+ from the endoplasmic reticulum, and subsequent addition of DPV caused no further change in [Ca2+]i. These data suggest that DPV-induced [Ca2+]i includes Ca release from the endoplasmic reticulum and Ca influx through store-operated calcium entry. Furthermore, DPV induced an increase in protein tyrosine phosphorylation, phosphorylation of Src and cortactin, actin remodeling, and altered transendothelial electrical resistance in BPAECs. These DPV-mediated effects were significantly attenuated by BAPTA (25 microM), a chelator of [Ca2+]i. Immunofluorescence studies reveal that the DPV-mediated colocalization of cortactin with peripheral actin was also prevented by BAPTA. Chelation of extracellular Ca2+ by EGTA had marginal effects on DPV-induced phosphorylation of Src and cortactin; actin stress fibers formation, however, affected EC barrier function. These data suggest that DPV-induced changes in [Ca2+]i regulate endothelial barrier function using signaling pathways that involve Src and cytoskeleton remodeling. PMID- 12882767 TI - Idiopathic and radiation-induced ocular telangiectasia: the involvement of the ATM gene. AB - PURPOSE: To investigate whether individuals, with no family history of ataxia telangiectasia (AT), in whom idiopathic or radiation-induced ocular telangiectasia developed are carriers of ATM gene mutations. METHODS: The ATM cDNA from lymphoblastoid cell lines established from 16 patients with idiopathic retinal or choroidal telangiectasia and 14 patients with radiation-induced telangiectasia after radiotherapy for age-related macular degeneration (AMD) was screened using the restriction endonuclease fingerprinting technique. The frequency of each detected variant was determined in the French population by either a mass spectrometry-based technique or variant-specific endonuclease digestion. RESULTS: Twenty-one ATM missense alterations, at 10 different sites, 8 of which would result in an amino acid substitution at a conserved position in the ATM protein were found. Four were novel changes, three of which were not detected in the 128 French control subjects screened. Eleven of 16 of the individuals with either idiopathic polypoidal choroidal vasculopathy or juxtafoveolar retinal telangiectasis and 6 of 14 individuals that had choroidal telangiectasis after radiotherapy for AMD carried ATM sequence variants. These latter six individuals had a significantly shorter delay time before the presentation of this vasculopathy compared with those individuals who had a wild type ATM (11.8 +/- 3.4 months vs. 17.5 +/- 4.5 months, P = 0.024). They had also received a lower average dose of X-rays, although this difference did not reach statistical significance (18.7 +/- 3.9 Gy vs. 23.7 +/- 5.6 Gy, P = 0.09). CONCLUSIONS: ATM missense variants could confer an AT-like phenotype and influence the formation of retinal and choroidal vascular abnormalities. PMID- 12882768 TI - Human lens thioredoxin: molecular cloning and functional characterization. AB - PURPOSE: To molecularly clone the human lens thioredoxin (TXN) gene, characterize the recombinant protein (rTrx1) and study the regulation expression of thioredoxin (Trx1) in human lens epithelial cells under oxidative stress. METHODS: The human TXN gene was cloned from a human lens cDNA library. Trx1 activity was measured by insulin reduction assay. For study of the upregulation of Trx1, 1.6 million human lens epithelial cells (HLE B3) were exposed to H(2)O(2) (0.1 mM) for 0, 5, 10, 15, 20, and 30 minutes. The cells were lysed with lysis buffer and used for Trx1 activity assay, Western blot analysis, and real time PCR. RESULTS: The sequence of the human lens TXN gene was identical with that of other human tissues. Recombinant Trx1 was sensitive to iodoacetic acid but showed strong resistance to oxidation (0.1 mM H(2)O(2)) at its approximate physiological protein level. Western blot analysis and assay of Trx activity revealed that Trx1 was expressed in HLE B3 cells and localized in epithelial, cortical, and nuclear regions of human and porcine lenses. In vivo oxidative stress of HLE B3 cells resulted in a 35% upregulation of the level of Trx1 protein after 10 minutes of H(2)O(2) treatment. Real-time PCR analysis showed an increase of approximately 50% in the level of Trx1 mRNA under the same oxidative stress conditions. CONCLUSIONS: The upregulation of Trx1 in HLE B3 cells under oxidative stress and the presence of Trx1 in the lens suggest that the thioredoxin system may be an effective defense system that protects the lens against oxidative stress. PMID- 12882769 TI - Mutations in the CHST6 gene in patients with macular corneal dystrophy: immunohistochemical evidence of heterogeneity. AB - PURPOSE: Macular corneal dystrophy (MCD) is an autosomal recessive disorder leading to severe visual impairment. The carbohydrate sulfotransferase 6 (CHST6) gene, which encodes the corneal N-acetylglucosamine 6-O-sulfotransferase on 16q22 has been identified as a causative gene for MCD. The purpose of this study was to identify mutations in CHST6 in Japanese patients with MCD and evaluate them by means of immunohistochemistry. METHODS: CHST6 was screened in 7 patients and 45 healthy control subjects. Genomic DNA was isolated, and the open reading frame (ORF) of CHST6 was amplified by polymerase chain reaction (PCR). PCR products were analyzed by direct sequencing and restriction enzyme digestion. Immunohistochemistry with a monoclonal anti-keratan sulfate (KS) antibody was performed on corneas of four patients from three families. RESULTS: Three novel mutations (P204Q, R205L, and R177H) and two previously reported mutations (R211W and A217T) were identified in the ORF of CHST6. P204Q, R205L, and R211W were found to be homozygous and R177H and A217T compound heterozygous with R211W on another allele. Immunohistochemistry revealed that R205L homozygous cornea had negative reactivity against the anti-KS antibody, representing type I MCD, and that R211W homozygous and R211W/A217T compound heterozygous corneas had negative or very weak reactivity in the stroma with antibody positive deposits, which were distinct from any previously reported types. CONCLUSIONS: Two mutations (homozygoous R211W and compound heterozygous R211W/A217T) should be subclassified immunohistochemically into new phenotypes of MCD. This heterogeneity could provide further insights into the pathogenesis of MCD. PMID- 12882770 TI - How sensitive to clinical change are ETDRS logMAR visual acuity measurements? AB - PURPOSE: To determine the sensitivity to change and specificity achieved when published test-retest variability (TRV) data are used to determine whether measured changes in ETDRS logarithm of the minimum angle of resolution (logMAR) visual acuity reflect true clinical change or are attributable to measurement error alone. METHODS: Various degrees of change in visual acuity were simulated in a group of normal subjects by adjusting test difficulty through manipulation of viewing distance. Sensitivity to simulated change and specificity were determined with change criteria derived from published Bland-Altman 95% ranges for TRV. RESULTS: The relationship between viewing distance and measured acuity was as predicted theoretically. Simulated acuity change of 0.2 logMAR (two lines of letters) or greater can be reliably distinguished from no change (both sensitivity and specificity >95%) with the ETDRS chart, but a change of 0.1 logMAR or less cannot. CONCLUSIONS: The use of 95% ranges for TRV to establish the smallest measured visual acuity change that can be reliably detected ensures a high specificity but does not take account of sensitivity. Use of change criteria derived from published 95% ranges results in a sensitivity of approximately 50% (assuming identical levels of TRV). Sensitivity may be improved by using a change criterion that is smaller than the minimum change sought, providing the change criterion is still at least as large as the 95% range for TRV, so that specificity is maintained. Reducing TRV allows smaller changes in acuity to be reliably detected. PMID- 12882771 TI - Uveal melanoma survival in Sweden from 1960 to 1998. AB - PURPOSE: To investigate the crude and relative survival rates in patients with uveal melanoma in Sweden during the period from 1960 to 1998. METHODS: A population-based national survey revealed 2997 cases of uveal melanoma in the Swedish Cancer Registry. The survival rates were calculated by the Hakulinen life table method, using relative survival as an estimate for deaths due to uveal melanoma. The excess mortality rates were calculated with confidence intervals for the first 15 years after diagnosis. Multivariate regression analysis was undertaken to evaluate the influence of gender, age, and calendar period on relative survival the first 5 years after diagnosis. The underlying causes of deaths in the patients with uveal melanoma, as found in the Cause of Death Registry were also investigated. RESULTS: Up to December 31, 1998, 2003 patients had died. The 5-year crude survival rate was 60.3% and the relative survival 70.1%. After 10 years, the rates were 42.5% and 59.4%, respectively. Significant excess mortality existed up to 5.5 years after diagnosis. In the multivariate model, younger age (P < 0.001) and later calendar period (P = 0.002), but not gender (P = 0.117), were associated with better relative survival. Deaths due to uveal melanoma were misclassified in the Cause of Death Registry in more than half of the cases. CONCLUSIONS: This study, covering more than 95% of the uveal melanoma cases in the Swedish population revealed an improvement in relative survival rates for patients with uveal melanoma over time and a significant excess mortality up to 5.5 years after diagnosis. PMID- 12882772 TI - Evaluation of sampling strategies for modeling survival of uveal malignant melanoma. AB - PURPOSE: To evaluate sampling strategies used to estimate survival after uveal malignant melanoma that exclude some patients who would be censored from the analysis. METHODS: Simulation was performed on a population-based data set of 133 patients who had an eye enucleated because of uveal melanoma. One thousand bootstrap samples of 80 patients were drawn, without replacement, according to three sampling strategies: a random draw (conventional strategy), a draw limited to patients who died of the tumor or survived at least 10 years without metastasis ("late-censoring" strategy), and a draw modified so that 40 patients died of melanoma and others survived at least 10 years without metastasis ("fifty fifty" strategy). The bias in the Kaplan-Meier analysis and Cox proportional hazards regression was quantified. RESULTS: The late-censoring strategy decreased the proportion of censored patients from 53% to 42%, whereas the fifty-fifty strategy assigned 50% of patients to this group. The former strategy overestimated mortality, the excess being 5.2% and 3.7% at 10 and 20 years, respectively. The latter strategy underestimated mortality, the bias being 1.6% and 4.6% at 10 and 20 years, respectively. The bias differed according to categories of explanatory variables so that the log-rank test statistic was inflated a median of 1.08 times (range, 0.73-1.87) and 1.14 times (range, 0.87 1.84), and the Wald chi(2) statistic of the Cox regression was inflated a median of 1.18 times (range, 0.79-2.13) and 1.16 times (range, 0.71-2.02), respectively, when the late-censoring and fifty-fifty strategies were applied. CONCLUSIONS: Sampling strategies that exclude on purpose a proportion of patients who would be censored produce biased statistics, because they violate assumptions of survival analysis. Only random sampling from an underlying population produces unbiased survival estimates. PMID- 12882773 TI - Repeatability of ETDRS visual acuity in children. AB - PURPOSE: To evaluate repeatability of the best corrected log minimum angle of resolution (MAR) Early-Treatment Diabetic Retinopathy Study (ETDRS) acuity in a group of 6- to 11-year old children with myopia. METHODS: Best corrected monocular visual acuity (VA) of a subset of children (n = 86) enrolled in the Correction of Myopia Evaluation Trial (COMET; mean spherical equivalent refractive error -2.35 D with no more than 1.25 D astigmatism) was measured at baseline and 1 month later with ETDRS logMAR charts. Children started with logMAR 0.4 (6/15 or 20/50) and read each letter on all subsequent lines until they missed all letters in 1 line. RESULTS: At baseline, the mean best corrected logMAR VA was 0.003 +/- 0.076 (6/6 or 20/20 +/- 3.8 letters) in the right eye and 0.008 +/- 0.059 (6/6 or 20/20 +/- 2.95 letters) in the left eye. The signed difference between VA measured at baseline and that measured at 1 month was not significantly different from zero in either eye. Repeatability was not associated with age, but a small, statistically significant association with gender was detected in the left eye, with boys approximately 2 letters more variable than girls. The kappa statistic (agreement within 1 line) was good to excellent. CONCLUSIONS: Based on the 95% limits of agreement, the criterion for a statistically significant change in VA is no more than +/-0.15 logMAR (or +/-8 letters). This value is similar to those reported for adults and indicates that logMAR VA provides a repeatable measure of acuity in children. PMID- 12882774 TI - A comparison of biological coatings for the promotion of corneal epithelialization of synthetic surface in vivo. AB - PURPOSE: To investigate the effect of a range of biological coatings on corneal epithelialization of a synthetic polymer surface in vivo. METHODS: Eight diverse biological factors (collagen I, collagen III, collagen IV, laminin, fibronectin, endothelial extracellular matrix, hyaluronic acid, and chondroitin sulfate) were coated individually onto the surface of polycarbonate membranes with a pore size of 0.1 micro m. The coated membranes were implanted on the anterior cornea of adult cats and were clinically assessed for rapidity and extent of and persistence of epithelial overgrowth. The membranes with persistent epithelial attachment were examined histologically by immunohistochemistry and routine light and electron microscopy. RESULTS: Collagen I, collagen IV, and laminin consistently enhanced migration and attachment of corneal epithelial cells in vivo. Multiple-layered epithelium over the collagen I-, collagen IV-, and laminin coated membranes was demonstrated histologically. The collagen I-coated membranes performed best, in that they showed greater stratification and differentiation of the epithelium. Formation of basement membrane and adhesion complexes over the collagen I-coated membranes was detected by immunohistochemistry and electron microscopy up to 9 weeks after implantation. Membranes coated by fibronectin, endothelial extracellular matrix, hyaluronic acid, and chondroitin sulfate did not support persistent epithelial overgrowth. Compromised biostability of these coatings was mostly likely associated with postsurgical reactions of the host corneal tissue. CONCLUSIONS: A biologically modified polymer can support migration and adhesion of corneal epithelial cells in vivo. The collagen I modified surface exhibited the most promising performance, both clinically and histologically. PMID- 12882775 TI - Mutation analysis of the carbohydrate sulfotransferase gene in Vietnamese with macular corneal dystrophy. AB - PURPOSE: Mutations in a new carbohydrate sulfotransferase gene (CHST6) encoding corneal N-acetylglucosamine-6-sulfotransferase (C-GlcNac-6-ST) have been identified as the cause of macular corneal dystrophy (MCD) in various ethnicities. This study was conducted to examine the CHST6 gene in Vietnamese with MCD. METHODS: Nineteen unrelated families, including 35 patients and 38 unaffected relatives were examined clinically. Blood samples were collected. Fifty normal Vietnamese individuals served as control subjects. Genomic DNA was extracted from leukocytes. Analysis of the CHST6 gene was performed with polymerase chain reaction and direct sequencing. Corneal buttons were studied histopathologically. RESULTS: A slit lamp examination revealed clinical features of MCD with gray-white opacities and stromal haze between. On histopathology, corneal sections showed positive staining with colloidal iron. Sequencing of the CHST6 gene revealed six homozygous and three compound heterozygous mutations. The homozygous mutations, including L59P, V66L, R211Q, W232X, Y268C, and 1067 1068ins(GGCCGTG) were detected, respectively, in two, one, eight, one, one, and two families. Compound heterozygous mutations R211Q/Q82X, S51L/Y268C, and Y268C/1067-1068ins(GGCCGTG) were identified, each in one family. A single heterozygous change at codon 76 (GTG-->ATG) was detected in family L, resulting in a valine-to-methionine substitution (V76M). None of these mutations was detected in the control group. CONCLUSIONS: Mutations identified in the CHST6 gene cosegregated with the disease phenotype in all but one family studied and thus caused MCD. Among these, the R211Q detected in 9 of 19 families may be the most common mutation in Vietnamese. These data also indicate that significant allelic heterogeneity exists for MCD. PMID- 12882776 TI - Production of fibronectin and tenascin isoforms and their role in the adhesion of human immortalized corneal epithelial cells. AB - PURPOSE: To study the production and deposition of fibronectin (Fn) isoforms and tenascin-C (Tn-C) by immortalized human corneal epithelial (HCE) cells and their integrin-dependent adhesion characteristics. METHODS: Indirect immunofluorescence with isoform-specific monoclonal antibodies (mAbs) was used to study extracellular matrix (ECM) protein composition and their integrin receptors in HCE cells. The synthesis of proteins was studied by Western blot analysis and adhesion by quantitative adhesion assay. RESULTS: HCE cells deposited fibrillar matrix containing extradomain EDA-Fn and sparser deposits of Onc-Fn, whereas Tn-C was deposited diffusely. EDA-Fn was present both in ECM and in culture medium, whereas Tn-C was present only in ECM. Fn-binding integrin (Int) alpha(5) subunit was present in subconfluent cells in focal adhesions (FAs) and matrix adhesions, whereas Int alpha(v)beta(5) was present in FAs in sparse cultures and as ringlike structures in denser cultures. Int alpha(v)beta(6) was colocalized with Int alpha(5) in FAs, only in cells adhering to growth substratum coated with Fn or Fn/Tn-C. Ints alpha(5)beta(1) and alpha(v)beta(6) mediated adhesion to Fn and Int alpha(v)beta(5) mediated adhesion to Vn, and both were inhibited by RGD peptide. The cells failed to adhere to Tn-C but adhered to Fn/Tn-C and were then more efficiently inhibited by the function-blocking integrin mAbs and RGD peptide. CONCLUSIONS: The results suggest corneal epithelial cells as the possible source for Fn isoforms and Tn-C in wound healing and pathologic conditions. The presence of Tn-C only in ECM suggest a vectorial deposition and adhesion experiments also indicate a role for Tn-C in Fn functioning. PMID- 12882777 TI - Predicting endothelial cell loss and long-term corneal graft survival. AB - PURPOSE: To evaluate a biexponential decay model for describing the loss of corneal endothelial cells with age as well as the increased loss of cells after cataract surgery and penetrating keratoplasty. METHODS: Data from previous studies were identified and the sum of two exponentials, d = p. exp(-at) + q. exp(-bt) (where d is cell density at time t, p and q are constants the sum of which is equal to the initial cell density, and a and b are exponential rate constants), fitted to each data set by a nonlinear least-squares algorithm. Goodness of fit was indicated by the residual standard deviation. Half times were calculated from the exponential rate constants. RESULTS: The model identified in each instance a rapid and a slow component to the cell loss. The half time for the slow component of the loss with age was 224 years, underlining the excess endothelial capacity in normal eyes. After surgery, the rapid component of the cell loss was probably due to surgical trauma and, after penetrating keratoplasty, cell-mediated rejection and other complications. The half times of the slow component were only 26 years after cataract surgery and 21 years after penetrating keratoplasty. DISCUSSION: The loss of endothelial cells followed a biexponential decay and could thus be described by a single equation. The half times of the slow component of the cell loss after surgery were substantially less than for the loss with age, indicating a markedly increased rate of cell loss that persisted for many years after surgery. A mechanism for this accelerated cell loss is suggested that involves a nonspecific, innate response initiated by the breakdown of the blood-ocular barrier. The model was used to calculate endothelial cell loss in the long term after penetrating keratoplasty and to predict when cell density would reach levels that are incompatible with maintenance of transparency and graft function. Thus, a rationale is presented for the setting of minimum donor cell densities by eye banks. PMID- 12882778 TI - Up-regulation of urokinase-type plasminogen activator in corneal epithelial cells induced by wounding. AB - PURPOSE: To investigate the possible role of urokinase-type plasminogen activator (uPA) in corneal epithelial wound healing by examining its expression both in the rabbit corneal epithelium in situ and in rabbit corneal epithelial (RCE) cells in vitro. METHODS: The rabbit cornea was subjected to mechanical wounding, and frozen sections of the tissue were subsequently prepared and subjected both to immunostaining with antibodies to uPA and to in situ zymography for the detection of PA activity. RCE cell monolayers were also subjected to scrape wounding, after which they were immunostained for uPA. The amounts of uPA protein in the culture medium and of uPA mRNA in cell lysates were also determined by enzyme-linked immunosorbent assay and by reverse transcription and real-time quantitative polymerase chain reaction analysis, respectively. RESULTS: Immunostaining and in situ zymography of the wounded cornea revealed that uPA was restricted to the leading edge of the migrating corneal epithelium. In contrast, tissue-type PA was expressed throughout the corneal epithelium. Scraping of RCE cell monolayers induced the expression of uPA in the migrating cells at the wound edge. The amount of uPA in the culture medium of RCE cells increased with the number of scrape wounds applied. Wounding also induced a time-dependent increase in the abundance of uPA mRNA in the cell monolayers. The migration of RCE cells was inhibited by antibodies to uPA. CONCLUSIONS: Mechanical wounding induces up regulation of uPA at both the protein and mRNA levels in corneal epithelial cells. uPA may thus contribute to epithelial cell migration during corneal epithelial wound healing. PMID- 12882779 TI - Serum albumin in mammalian cornea: implications for clinical application. AB - PURPOSE: To compare the abundance and spatial distribution of serum albumin in the mouse and bovine cornea. METHODS: Serum albumin from cornea was separated from transketolase by SDS-PAGE (+/-dithiothreitol [DTT]) and identified by peptide sequencing and immunoblot analyses. The fractional content of serum albumin was determined in water-soluble extracts of cornea by imaging analyses after SDS-PAGE. Serum albumin was localized in cornea by immunohistochemistry and by SDS-PAGE analyses of samples from separated epithelium and stroma. RESULTS: SDS-PAGE (-DTT) resolved mouse serum albumin and transketolase and indicated that serum albumin was 13% of the water-soluble protein in whole mouse corneas. By contrast, corneal epithelial fractions contained little (<1%) serum albumin. Immunohistochemistry indicated that mouse serum albumin was present throughout the stroma between collagen lamellae. Immunohistochemical analyses of bovine cornea yielded similar results. In addition, immunohistochemistry for serum albumin revealed positive staining in a small number of basal epithelial cells next to Bowman's membrane, and greater staining in the anterior-peripheral stroma as well as immediately adjacent to Descemet's membrane. CONCLUSIONS: Mouse and bovine cornea have a similar content and spatial distribution of serum albumin. The appreciable serum albumin in the cornea documented here and elsewhere raise the possibility that it contributes to the physiological or optical functions of the cornea. Moreover, serum albumin's ability to bind drugs suggests that mice corneas could be exploited to study drug-serum albumin interactions in vivo and to test the usefulness of serum albumin as a drug carrier for corneal disorders. PMID- 12882780 TI - Rat corneal allograft survival prolonged by the superantigen staphylococcal enterotoxin B. AB - PURPOSE: The purpose of this study was to determine the optimal conditions for prolonging corneal allograft survival by inducing anergy with the superantigen staphylococcal enterotoxin B (SEB). METHODS: A rat model of penetrating keratoplasty, whereby Fisher344 donor corneas are implanted into Lewis recipients, was used to evaluate the effects of SEB on inhibiting immune-mediated allograft rejection. To induce anergy, SEB was injected into the peribulbar space of Lewis rats. Furthermore, histopathology and immunofluorescent staining were used to examine the levels of infiltrating CD4(+) and CD8(+) T lymphocytes and NK1.1(+) lymphocytes. RESULTS: By administering SEB, at doses of 90 or 120 micro g/kg 7 days before and after keratoplasty, we suppressed the episode of corneal graft rejection for a median of 12 and 30 days, respectively. In contrast, rejection was observed when 30 or 60 micro g/kg of SEB was administered. After SEB injections, lymphocyte infiltration into the corneal grafts was reduced, and the expression of NK1.1(+) lymphocytes was enhanced, suggesting that anergy may be occurring. Also, there were no differences in the number of infiltrating CD4(+) and CD8(+) T lymphocytes cells between the control group and groups injected with 30 and 120 micro g/kg SEB on postoperative days 10 and 30. CONCLUSIONS: Inducing anergy with the superantigen SEB prolonged corneal graft survival in a rat model of penetrating keratoplasty. Therefore, these results support the possibility of prolonging corneal allograft survival in a clinical setting by preventing immune-mediated rejection through the administration of the superantigen SEB. PMID- 12882781 TI - Presence and localization of neurotrophins and neurotrophin receptors in rat lacrimal gland. AB - PURPOSE: To determine which of the neurotrophins (NTs)-nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT3), and neurotrophin 4/5 (NT4)-and their receptors (NTrs), TrkA, TrkB, TrkC, and p75, are present in the adult rat lacrimal gland. METHODS: RT-PCR was performed on RNA isolated from male rat lacrimal gland, using oligonucleotides specific to each NT and NTr. The presence of NT and NTr protein, was determined by Western blot analysis of lacrimal gland homogenate or membranes. The location of NTs and NTrs was determined by immunofluorescence histochemistry. Western blot analyses and immunofluorescence microscopy were performed using primary rabbit polyclonal antibodies raised against NTs and NTrs. RESULTS: RT-PCR showed positive bands at the appropriate sizes for NGF, BDNF, NT3, and NT4, and for the receptors TrkA, TrkB, TrkC, and p75. Western blot analysis confirmed these results, showing that the lacrimal gland expresses NGF, BDNF, NT3, and NT4 as well as the NTrs TrkA, TrkB, and TrkC and the p75 protein. NGF, BDNF, NT3, and NT4 were localized in the lacrimal gland acini with differing cellular distributions, whereas TrkA, TrkB, and TrkC, were localized in myoepithelial cell and ductal cell membranes. The protein p75 was expressed only on myoepithelial cell membranes. CONCLUSIONS: Members of the neurotrophin family of growth factors and their receptors are present in rat lacrimal gland, which suggests a role for NTs and their receptors in the lacrimal gland. PMID- 12882782 TI - Maldevelopment of convergence eye movements in macaque monkeys with small- and large-angle infantile esotropia. AB - PURPOSE: To describe symmetric convergence eye movements evoked by disparity and/or accommodative cues in esotropic macaque monkeys, with the goal of determining whether these animals have the vergence deficits found in humans with esotropia. METHODS: Physical far and near targets were used to evoke large (approximately 8 degrees) symmetric convergence eye movements in four adult macaque monkeys (two with strabismus, two normal), using positive-feedback rewards. One strabismic monkey had infantile-onset, small-angle esotropia (small eso approximately 2 degrees) induced by alternating occlusion from birth to age 9 months. The other strabismic monkey had naturally occurring, large-angle (approximately 25 degrees) infantile-onset esotropia (large-eso). Visual acuity was normal in each eye as measured by spatial sweep visually evoked potentials (VEPs). Eye movements were recorded using magnetic search coils. RESULTS: When viewing binocularly, both normal monkeys exhibited accurate, stereotyped symmetric convergence movements that achieved 87% to 96% of the required change in vergence angle by the end of the initial movement. In contrast, the small-eso monkey's convergence response when viewing binocularly was variable, strikingly asymmetric, usually accompanied by a disjunctive saccade, and subnormal, achieving only 56% of required vergence. The convergence response of the large eso monkey was also asymmetric and weak, achieving 18% of the required vergence and employing conjugate saccades to refixate the near target. Monocular viewing (i.e., accommodative vergence) caused substantial reductions in both convergence amplitudes and velocities in the normal monkeys, but had a minor effect on the vergence behavior of the strabismic animals. CONCLUSIONS: Monkeys with small- and large-angle infantile esotropia have striking maldevelopments of binocular (disparity-driven) convergence and use accommodative vergence and saccades to refixate near targets. Their vergence behavior resembles that in esotropic humans. The maldevelopment may be explained in large part by the paucity of binocular connections recently described in the visual cortex of esotropic macaques. PMID- 12882783 TI - Optical tomography-measured retinal nerve fiber layer thickness in normal latinos. AB - PURPOSE: To measure retinal nerve fiber layer (RNFL) thickness in normal Latinos using optical coherence tomography (OCT). METHODS: Three hundred twelve Latino participants, aged 40 years or more, underwent a detailed ophthalmic examination, including measurement of visual acuity, intraocular pressure, visual field perimetry, and stereoscopic optic disc photography. None of the participants had any evidence of ocular hypertension, glaucoma, or other ocular disease. Nine scans were performed on one eye of each participant by optical coherence tomography (OCT Model 2000; Carl Zeiss Meditec, Dublin, CA, software version A 6.1): three circumferential peripapillary scans and six radial scans of the macula. The average RNFL thicknesses in the peripapillary region and macula were measured. A paired t-test and linear regression analyses were used to analyze the data. RESULTS: The mean age of the participants was 52 years (range, 40-79 years). The average peripapillary RNFL thickness 1.74 mm from the center of the disc was 132.7 +/- 14.4 micro m, and the average macular RNFL thickness was 44.8 +/- 14.8 micro m. The average macular retinal thickness was 173 +/- 28.5 micro m. The average peripapillary RNFL thickness in the four quadrants was as follows: superior 157.7 +/- 17.8 micro m, nasal 109.3 +/- 19.1 micro m, inferior 159.8 +/- 18.9 micro m, and temporal 102.5 +/- 19.0 micro m. There were no gender-related differences in macular or peripapillary RNFL thickness (P = 0.12 and P = 0.35, respectively). The average macular and peripapillary RNFL thickness was thinner in older Latinos than in younger Latinos (P = 0.04 and P = 0.0001, respectively). CONCLUSIONS: Regional and age-related differences in the peripapillary and macular RNFL thickness should be considered when diagnosing and monitoring individuals with diseases that affect the RNFL. PMID- 12882784 TI - Antigenic specificity of immunoprotective therapeutic vaccination for glaucoma. AB - PURPOSE: To investigate the antigenic specificity of the immune neuroprotective mechanism that can protect retinal ganglion cells (RGCs) against death caused by high intraocular pressure (IOP). METHODS: A unilateral increase in IOP was induced in rats by argon laser photocoagulation of the episcleral veins and limbal plexus. Rats with high IOP were immunized with glatiramer acetate (Cop-1, a synthetic copolymer) or with myelin-derived or uveitogenic peptides. When the steroid drug methylprednisolone was used, it was administered intraperitoneally every other day for 12 days. RESULTS: Vaccination with myelin-derived peptides that reside in the axons failed to protect RGCs from death caused by high IOP. In contrast, IOP-induced RGC loss was reduced by vaccination with R16, a peptide derived from interphotoreceptor retinoid-binding protein, an immunodominant antigen residing in the eye. The benefit of protection against IOP-induced RGC loss outweighed the cost of the monophasic experimental autoimmune uveitis (EAU) that transiently developed in a susceptible rat strain. Treatment with methylprednisolone alleviated the disease symptoms, but caused further loss of RGCs. Cop-1 vaccination was effective in both EAU-resistant and EAU-susceptible strains. CONCLUSIONS: To benefit damaged neurons, immune neuroprotection should be directed against immunodominant antigens that reside in the site of damage. In a rat model of high IOP, RGCs can benefit from vaccination with peptides derived from proteins that are immunodominant in the eye but not from myelin-associated proteins. This suggests that the site of primary degeneration in IOP-induced RGC loss is in the eye. Cop-1 vaccination apparently circumvents the site-specificity barrier and provides protection without risk of inducing autoimmune disease. PMID- 12882785 TI - Racial differences in the association between optic disc topography and early glaucoma. AB - PURPOSE: To determine the structural characteristics of the optic disc that are associated with early glaucoma in African Americans and whites and whether these characteristics differ between the races. METHODS: Parameters of optic disc topography from 260 African American eyes and 193 white eyes were included in the analysis. One hundred forty-four eyes of African Americans and 109 eyes of normal white subjects were used as a control group. Logistic regression was used to calculate the association between early glaucoma, defined by the visual field, and cup, rim, and disc margin confocal scanning laser ophthalmoscopic (CSLO) parameters, using odds ratios at binary cut points. The cup, rim, and disc margin parameters identified as being independently associated with glaucoma in these reduced models were then included in a single multivariate model. Optic disc area was included in the analysis at each level of the model. This approach was used for the total study group and then separately for the African American and white groups. RESULTS: When accounting for difference in optic disc area, rim area had the highest independent association with early glaucoma in both groups, but this association was lower in African Americans (odds ratio [95% confidence interval]: 1.63 [1.12-2.36]) than in whites (odds ratio: 4.74 [2.18-10.28]). Additional independently associated parameters included cup shape, maximum elevation along the contour line, and the temporal-to-inferior contour line modulation ratio in whites and cup shape and the temporal-to-superior contour line modulation ratio in African Americans. CONCLUSIONS: Structural characteristics of the optic disc that are best associated with early glaucoma included cup shape and rim area in both groups, but with a less pronounced association in African Americans. In addition, several other race-specific parameters that were independently associated with early glaucoma differed significantly between African Americans and whites. These race-specific differences were independent from the effect of optic disc area. PMID- 12882786 TI - A model to study differences between primary and secondary degeneration of retinal ganglion cells in rats by partial optic nerve transection. AB - PURPOSE: To use a rat model of optic nerve injury to differentiate primary and secondary retinal ganglion cell (RGC) injury. METHODS: Under general anesthesia, a modified diamond knife was used to transect the superior one third of the orbital optic nerve in albino Wistar rats. The number of surviving RGC was quantified by counting both the number of cells retrogradely filled with fluorescent gold dye injected into the superior colliculus 1 week before nerve injury and the number of axons in optic nerve cross sections. RGCs were counted in 56 rats, with 24 regions examined in each retinal wholemount. Rats were studied at 4 days, 8 days, 4 weeks, and 9 weeks after transection. The interocular difference in RGCs was also compared in five control rats that underwent no surgery and in five rats who underwent a unilateral sham operation. It was confirmed histologically that only the upper optic nerve had been directly injured. RESULTS: At 4 and 8 days after injury, superior RGCs showed a mean difference from their fellow eyes of -30.3% and -62.8%, respectively (P = 0.02 and 0.001, t-test, n = 8 rats/group), whereas sham-operation eyes had no significant loss (mean difference between eyes = 1.7%, P = 0.74, t-test). At 8 days, inferior RGCs were unchanged from control, fellow eyes (mean interocular difference = -4.8%, P = 0.16, t-test). Nine weeks after transection, inferior RGC had 34.5% fewer RGCs than their fellow eyes, compared with 41.2% fewer RGCs in the superior zones of the injured eyes compared with fellow eyes. Detailed, serial section studies of the topography of RGC axons in the optic nerve showed an orderly arrangement of fibers that were segregated in relation to the position of their cell bodies in the retina. CONCLUSIONS: A model of partial optic nerve transection in rats showed rapid loss of directly injured RGCs in the superior retina and delayed, but significant secondary loss of RGCs in the inferior retina, whose axons were not severed. The findings confirm similar results in monkey eyes and provide a rodent model in which pharmacologic interventions against secondary degeneration can be tested. PMID- 12882787 TI - Evaluation of anti-TGF-beta2 antibody as a new postoperative anti-scarring agent in glaucoma surgery. AB - PURPOSE: Postoperative subconjunctival wound healing remains the commonest cause of late bleb failure after glaucoma filtration surgery. This study was undertaken to investigate whether the human monoclonal antibody that neutralizes transforming growth factor-beta2 (CAT-152; lerdelimumab) could be used as a postoperative agent to prevent scarring after glaucoma surgery and compared it with 5-fluorouracil (5-FU), to benchmark its potential clinical benefit. METHODS: In a randomized, controlled, masked-observer study, after modified glaucoma surgery, 48 rabbits were randomly allocated to receive a postoperative course of seven subconjunctival injections of CAT-152 (1 mg/mL), 5-FU (50 mg/mL), or no treatment. Bleb characteristics, the presence of subconjunctival drainage, and local reaction to treatment were assessed. Animals were killed on days 10, 21, and 30. Immunohistochemistry, histologic staining and electron microscopy were performed to demonstrate the mechanism of CAT-152-mediated effects on the extracellular matrix. RESULTS: CAT-152 significantly improved surgical outcome (log rank test, P < 0.001) and reduced subconjunctival collagen deposition (P < 0.01) compared with 5-FU and control. Median bleb survival was increased in the CAT-152 group (23.5 days) compared with the 5-FU (20 days) and control (16 days) treatment groups. CAT-152 treatment improved bleb morphology (P < 0.05) and was well tolerated. 5-FU prolonged the duration of corneal epitheliopathy (P < 0.01). CONCLUSIONS: Postoperative administration of CAT-152 significantly improved surgical outcome, reduced subconjunctival scarring, and minimized the risk of corneal side effects compared with the anti-scarring agent 5-FU. These findings suggest that CAT-152 may offer therapeutic benefit as a postoperative agent to prevent subconjunctival scarring after glaucoma filtration surgery. PMID- 12882788 TI - The critical role of IFN-gamma in experimental coronavirus retinopathy. AB - PURPOSE: Experimental coronavirus retinopathy (ECOR) is an animal model of progressive retinal disease that is first manifest as an acute retinal inflammation followed by chronic, immune-associated retinal degeneration in genetically susceptible, BALB/c mice. In retinal degeneration-resistant CD-1 mice, only the acute infection is seen. In the present study, interferon (IFN) gamma production during ECOR was studied and its role evaluated in the clearance of infectious virus from the retina. METHODS: BALB/c, CD-1, and IFN-gamma deficient (IFN-gamma gko) mice were inoculated with the JHM strain of murine coronavirus by the intravitreal route. Mouse eyes were evaluated for infiltrating cells and major histocompatibility complex (MHC) expression by immunocytochemical staining. Isolated retinas were analyzed for IFN-gamma mRNA by RT-PCR, and sera were evaluated for IFN-gamma protein by ELISA assays. RESULTS: Virus infection in BALB/c mice was associated with an increase in the incidence and levels of systemic IFN-gamma. Moreover, IFN-gamma mRNA was detected within the retinas of infected animals during the acute phase of the disease but was not detected in normal or mock-injected animals. IFN-gamma mRNA was detected at the time of T cell infiltration, and earlier studies have shown that this is temporally related to granzyme B gene expression and the clearance of infectious virus from the retina. Retinal IFN-gamma mRNA was also associated with the upregulation of MHC class I and II molecules within the retina. When this infection occurred in IFN gamma gko mice, the virus was unchecked, and the infection led to death. CONCLUSIONS: These studies indicate that generation of IFN-gamma by cells infiltrating the retina is an essential part of an immune mechanism responsible for noncytolytic clearance of infectious virus from the retina. PMID- 12882789 TI - Analysis of Pseudomonas aeruginosa corneal infection using an oligonucleotide microarray. AB - PURPOSE: To compare the early gene expression pattern of normal versus Pseudomonas aeruginosa-infected corneas in resistant (cornea heals) versus susceptible (cornea perforates) mice. METHODS: A microarray analysis of normal versus postinfection (PI) day 1 BALB/c and B6 corneas was performed with a murine gene microarray. Real-time RT-PCR was used to confirm the microarray pattern selectively. RESULTS: The 1257 regulated transcripts detected were organized into nine clusters by a self-organizing map (SOM) algorithm according to their different behavior in each mouse group. At least three groups of genes associated with a CD4(+) T-cell type-1 (Th1) immune response and three clusters linked with a type-2 T-cell (Th2) response were identified. Biological categorization revealed that the cornea of B6 mice showed a dominant type-1-like immune response profile, whereas BALB/c mice showed a dominant type-2-like profile. In addition, expression of several genes that promote apoptosis (e.g., caspase-9) was upregulated in BALB/c mouse cornea, whereas genes with apoptosis-inhibiting activity (e.g., BCL2) were significantly upregulated in B6 mouse cornea. The infected cornea of BALB/c mice also showed increased gene expression of factors associated with matrix remodeling and tissue repair (e.g., tissue inhibitor of matrix metalloproteinase [TIMP-2] and epidermal growth factor [EGF]) and/or bacterial killing (e.g., inducible nitric oxide synthase [iNOS]). CONCLUSIONS: The data provide new insight into biological processes involved in Pseudomonas aeruginosa keratitis and confirm that B6 mice are Th1 and BALB/c mice are Th2 cytokine responsive to bacterial antigen early after challenge with P. aeruginosa. PMID- 12882790 TI - Effects of IL-4 on conjunctival fibroblasts: possible role in ocular cicatricial pemphigoid. AB - PURPOSE: Increased stromal accumulation of macrophages and submucosal fibrosis due to excessive accumulation of collagens are central histologic features in ocular cicatricial pemphigoid (OCP). Interleukin (IL)-4 plays an important role in both the inflammatory and fibrotic events in several human and experimental diseases. In the present study, the possible role of IL-4 in the pathogenesis of OCP was investigated. METHODS: Biopsy specimens from the conjunctivae of 10 patients with OCP and 5 normal subjects were studied for the expression of IL-4 by immunohistochemistry. The expression level of IL-4 was also examined in conjunctival fibroblasts of normal control subjects and patients with OCP. The effects of IL-4 in the induction of inflammatory and fibrogenic molecules was studied in IL-4-treated conjunctival fibroblasts, and the expression levels of macrophage colony-stimulating factor (m-CSF), heat shock protein (HSP)-47 and type I collagen was determined by quantitative real-time PCR. The level of IL-4 was also measured by enzyme-linked immunosorbent assay (ELISA) in serum samples obtained from patients with OCP during active stage and remission and were compared with the levels in control sera. RESULTS: Compared with the weak expression of IL-4 in the normal conjunctival sections, an increased expression of IL-4 was noted in conjunctival sections of patients with OCP. A similar increase in the expression of IL-4 was also detected in fibroblasts isolated from conjunctiva of patients with OCP, compared with control fibroblasts. Real-time PCR and ELISA detected a significantly increased level of m-CSF, at both the mRNA and protein levels in IL-4-stimulated cells. Similarly, IL-4 treatment resulted in the induction of type I collagen and collagen-binding HSP47 by conjunctival fibroblasts, as detected by real-time PCR. However, no apparent changes in the levels of IL-4 were detected by ELISA in serum samples of patients with OCP and control subjects. CONCLUSIONS: Increased conjunctival expression of IL-4 may play an important role in the regulation of local accumulation of macrophages (by inducing m-CSF), and matrix accumulation (by inducing HSP47 and collagen) during conjunctival scarring in patients with OCP. IL-4, therefore, may augment or enhance both conjunctival inflammatory and subsequent fibrotic responses in patients with OCP. PMID- 12882791 TI - Effects of mannose on Acanthamoeba castellanii proliferation and cytolytic ability to corneal epithelial cells. AB - PURPOSE: Acanthamoeba trophozoites express a mannose binding receptor that facilitates adhesion of trophozoites to mannosylated proteins on corneal epithelial cells. This study was undertaken to determine the role that mannose stimulation has in the amoeba's growth, secreted products, and ability to desquamate the corneal epithelium. METHODS: Acanthamoeba castellanii trophozoites were grown in peptone-yeast extract glucose (PYG) and PYG with 100 mM methyl alpha-D-mannopyranoside or galactose. The proliferation of trophozoites and cysts was examined by optical density and direct counts. The molecular weight of the mannose-stimulated protein was examined by SDS/PAGE. The cytolytic protein was purified by fast protein liquid chromatography (FPLC) size exclusion and ionic exchange and then tested for cytopathic effect (CPE) and collagenolytic activity in vitro. Collagenolytic activity was examined by zymography. Proteases and protease inhibitors were used to characterize the nature of the cytolytic protein. RESULTS: Methyl alpha-D-mannopyranoside inhibited the growth of A. castellanii by 50% (P < 0.05) and concomitantly induced a threefold increase in the formation of cysts. SDS-PAGE analysis revealed a mannose-induced protein of approximately 133 kDa (MIP-133). The MIP-133 protein was found to be highly cytolytic against corneal epithelial cells, but not human intestinal epithelial cells and also degraded collagen in vitro. Serine protease inhibitors abrogated both CPE and collagenolytic activity of the MIP-133 protein (P < 0.001). CONCLUSIONS: The results suggest that binding of trophozoites to mannosylated proteins on the corneal surface induces A. castellanii to secrete a approximately 133-kDa serine protease that kills both human and hamster corneal epithelium and degrades collagen. PMID- 12882792 TI - Differential regulation of ENA-78 and GCP-2 gene expression in human corneal keratocytes and epithelial cells. AB - PURPOSE: To determine whether interleukin (IL)-1alpha- and tumor necrosis factor (TNF)-alpha-stimulated human corneal epithelial cells (HCECs) and human corneal keratocytes (HCKs) produce the alpha-chemokines epithelial cell-derived neutrophil attractant (ENA)-78 and granulocyte chemotactic protein (GCP)-2. METHODS: Cultures of HCECs and HCKs were stimulated with either human recombinant IL-1alpha or TNF-alpha. At selected times after stimulation, culture supernatants were harvested and assayed for ENA-78 and GCP-2 by enzyme-linked immunosorbent assay. RNA was extracted from cell cultures to measure steady state levels of intracellular ENA-78 and GCP-2 pre-mRNA and mRNA by the reverse transcription polymerase chain reaction. RESULTS: Exposure of HCECs to either IL-1alpha or TNF alpha stimulated a more than 4.5-fold increase in ENA-78 RNA and protein synthesis without stimulating a significant increase in either GCP-2 RNA synthesis or protein production. Exposure of HCK to IL-1alpha stimulated a 10 fold increase in ENA-78 and GCP-2 RNA synthesis and a more than 300-fold increase in ENA-78 and GCP-2 protein production. In contrast, exposure of keratocytes to TNF-alpha significantly enhanced ENA-78 RNA synthesis, resulting in a more than 68-fold increase in ENA-78 protein synthesis without significantly enhancing either GCP-2 gene expression or protein secretion. CONCLUSIONS: ENA-78 gene expression is significantly enhanced in both HCECs and HCKs in response to either IL-1alpha or TNF-alpha stimulation. In contrast, GCP-2 synthesis is only inducible in IL-1alpha-stimulated HCKs. The results suggest that GCP-2 gene expression is more tightly regulated in diseased or injured corneal tissue than is ENA-78 gene expression. PMID- 12882793 TI - Mechanisms and physiological significance of the transport of the glutathione conjugate of 4-hydroxynonenal in human lens epithelial cells. AB - PURPOSE: To study the mechanism and physiological significance of the transport of the glutathione (GSH) conjugate of 4-hydroxynonenal (4-HNE) from lens epithelial cells. METHODS: HLE B-3 cells were treated with [(3)H] 4-HNE and efflux of its GSH conjugate, [(3)H] GS-HNE, into the medium was quantitated and characterized by HPLC and mass spectrometry. Inside-out vesicles (IOVs) were prepared from HLE B-3 cell membranes. The kinetics of adenosine triphosphate (ATP)-dependent uptake of GS-HNE and dinitrophenyl S-glutathione (DNP-SG) by these IOVs and inhibition of GS-HNE uptake by anti-RLIP76 IgG was studied. Localization of RLIP76 was studied by immunogold electron microscopy and kinetics of the adenosine triphosphatase (ATPase) activity of purified RLIP76 was determined. 4-HNE-induced apoptosis was compared in HLE-B3 cells coated with anti RLIP76 IgG or preimmune IgG, by caspase activation assay. RESULTS: The results showed the presence of RLIP76 in plasma membranes of HLE B-3 cells and that it mediated ATP-dependent transport of GS-HNE as well as of DNP-SG. The transport was saturable with respect to GS-HNE (K(m) = 8.4 micro M), DNP-SG (100 micro M) as well as to ATP (K(m) 0.45 mM) and was sensitive to temperature and osmolarity of the medium. Anti-RLIP76 IgG inhibited approximately 65% of the transport of GS HNE and DNP-SG, indicating that most of the transport was mediated by RLIP76. Compatible with its transport function, the EGTA- and ouabain-insensitive ATPase activity of purified RLIP76 was stimulated by DNP-SG and GS-HNE. 4-HNE-induced caspase activation in HLE-B3 cells was exacerbated when the transport of GS-HNE from these cells was blocked by anti-RLIP76 IgG. CONCLUSIONS: RLIP76 provides a defense against the deleterious effects of 4-HNE by transporting GS-HNE and can modulate apoptotic signaling by regulating the intracellular concentrations of 4 HNE. PMID- 12882794 TI - A new model of posterior capsule opacification in rodents. AB - PURPOSE: To describe a new model of posterior capsule opacification (PCO) in rodents METHODS: An extracapsular lens extraction (ECLE), by continuous curvilinear capsulorrhexis and hydrodissection, was performed in 42 consecutive Brown Norway rats. Animals were killed at 0, 6, and 24 hours and 3, 7, and 14 days after surgery. Eyes were enucleated and processed for light microscopy and immunohistochemistry. RESULTS: In 34 (81%) of the animals the operated eye appeared well healed before death, with a clear cornea and a well-formed anterior chamber. In eight (19%) there was no view of anterior segment structures because of hyphema, fibrin, or corneal opacification. PCO was clinically evident 3 days after ECLE and was present in all animals at 2 weeks. Immediately after ECLE, lens epithelial cells (LECs) were present in the inner surface of the anterior capsule and lens bow. Twenty-four hours after surgery, LECs started to migrate toward the center of the posterior capsule. At 3 days, multilayered LECs, some spindle shaped, were present throughout the lens capsule. Capsular wrinkling was apparent. Lens fibers and Soemmering's ring were observed in all animals 14 days after surgery, indicating some degree of cellular differentiation. Activated macrophages were present in greater numbers at 3 and 14 days after surgery (P < 0.05), when proliferation and migration of LECs appeared to be greatest, and lens fiber differentiation was evident, respectively. CONCLUSIONS: In rodents PCO occurs after ECLE and is associated with low-grade inflammation, mostly of mononuclear macrophages. Although no intraocular lens implantation was performed, this model appears to be valuable for studying the sequence of events that leads to PCO after cataract surgery and the extracellular matrix cues that promote lens fiber differentiation. PMID- 12882795 TI - Expression patterns for glucose transporters GLUT1 and GLUT3 in the normal rat lens and in models of diabetic cataract. AB - PURPOSE: To determine whether the expression levels and cellular distribution of the facilitative glucose transporters GLUT1 and -3 undergo changes in the hyperglycemic lens. METHODS: Hyperglycemia was induced in vivo by injecting rats with streptozotocin or in vitro by culturing lenses in the presence of 50 mM glucose. Northern blot analysis and quantitative RT-PCR were used to detect changes in GLUT1 and -3 transcript levels, and Western blot analysis was used to monitor changes in GLUT3 protein expression levels in diabetic rats. Immunocytochemistry was used to map the cellular distribution of GLUT3 in normal and hyperglycemic lenses. RESULTS: GLUT1 and -3 were found to be differentially expressed in the epithelial and fiber cells, respectively. In the fiber cells, the distribution of GLUT3 protein changed as a function of fiber cell differentiation. In young differentiating fiber cells, GLUT3 was mainly found in the cytoplasm, but with increasing depth into the lens became inserted into the narrow sides of older fiber cells, before becoming completely dispersed around the entire membrane of the oldest fiber cells. Hyperglycemia had similar effects on tissue damage and transporter expression in both the in vitro and in vivo models. Tissue damage was characterized by an initial local cell swelling that with prolonged insult gradually spread and resulted in the creation of large areas of tissue liquefaction. Northern blot analysis and quantitative RT-PCR showed that transcript for GLUT3 but not GLUT1 was upregulated under hyperglycemic conditions. This increase in GLUT3 expression was confirmed at the protein level by both Western blot analysis and immunocytochemistry. In hyperglycemic lenses, GLUT3 antibody labeling was localized to the region of tissue liquefaction. CONCLUSIONS: GLUT3 in the lens exhibits dynamic changes in expression levels and cellular localization as a function of fiber cell differentiation and hyperglycemia. In the lens cortex, regions of GLUT3 overexpression and hyperglycemic tissue damage overlap, suggesting a functional relationship. PMID- 12882796 TI - The effect of up- and downregulation of MnSOD enzyme on oxidative stress in human lens epithelial cells. AB - PURPOSE: Gene knockouts serve as useful experimental models to investigate the role of antioxidant enzymes in protection against oxidative stress in the lens. In the absence of gene knockout animals for Mn-containing superoxide dismutase (MnSOD), the effect of this enzyme on oxidative stress was investigated in a human lens epithelial cell line (SRA 01/04) in which the enzyme was up- or downregulated by transfection with sense and antisense expression vectors for MnSOD. METHODS: Human lens epithelial cells (SRA 01/04) were transfected with plasmids containing sense and antisense human cDNA for MnSOD. The enzyme levels were determined by Western blot analysis and immunocytochemistry. The protective effect of the enzyme against the cytotoxicity of H(2)O(2) was evaluated by cell viability, DNA strand breaks, and morphologic changes observed by transmission electron microscopy. In addition, the protective effect of this enzyme against photochemically induced stress and UVB irradiation in cells was assessed by measuring the damage of cellular DNA. RESULTS: The MnSOD level in upregulated cells was three times higher than in downregulated cells, and the enzyme surrounded the nucleus. Cells with elevated enzyme levels were more resistant to the cytotoxic effect of H(2)O(2) with greater cell viability. MnSOD-deficient cells showed dramatic mitochondrial damage when exposed to 50 micro M H(2)O(2) for 1 hour. Similarly, oxidative challenge by H(2)O(2), photochemically generated reactive oxygen species, or UVB irradiation produced greater DNA strand breaks in MnSOD-deficient cells than in those in which the enzyme was upregulated. CONCLUSIONS: These findings demonstrate the protective effect of MnSOD in antioxidant defense of cultured lens epithelial cells. This approach to modulating the enzyme level in cultured cells provides a new experimental model for study of the role of antioxidant enzymes in the lens. PMID- 12882797 TI - Lenticular oxygen toxicity. AB - PURPOSE: To investigate the possible toxic effect of oxygen on lenses in an organ culture. METHODS: Bovine lenses were exposed to four different combinations of ambient pressure and oxygen concentration in an organ culture throughout a 7-day period. Lens transparency, histology, enzymatic activities, and photomicrographs were compared in study and control groups. RESULTS: No differences were observed between study and control lenses in all measured parameters in a group subjected to a single exposure of 100% oxygen under increased (i.e., hyperbaric) ambient conditions and a group exposed repeatedly to high ambient pressure and normal oxygen partial pressure. Decreased lenticular transparency and enzymatic activities along with structural changes were observed in lenses exposed repeatedly to 100% oxygen concentration under both normal and increased ambient pressures. The observed changes were oxygen-load-dependent: the higher the oxygen partial pressure and the longer the time of exposure, the more severe the changes observed. Optical and structural changes in the lens occurred in a centripetal orientation: the greater the oxygen load, the more central the damage. CONCLUSIONS: High oxygen load has a toxic effect on bovine lenses in organ culture. These effects appear to be cumulative: the higher the oxygen partial pressure and the greater the number of exposures, the more severe the changes observed in the lenses. Changes marking toxicity follow the route of oxygen diffusion into the lens, from the periphery to the center. Cautious interpretation of the results may indicate a role of oxygen (and/or its derivatives) in human cataract formation. PMID- 12882798 TI - Expression of matrix metalloproteinases and their inhibitors in human trabecular meshwork cells. AB - PURPOSE: Matrix metalloproteinases (MMPs) are involved in trabecular meshwork (TM) extracellular matrix metabolism and have been shown to increase aqueous outflow facility. The purpose of this study was to characterize effects of cytokines, a phorbol ester, and prostanoids on the expression of MMP-1, -2, -3, and -9 and tissue inhibitors of metalloproteinases (TIMP)-1 and -2 in cultured human TM cells. METHODS: Five human TM cell strains were treated with selected compounds. Levels of proMMPs and TIMPs in cell media were quantified by ELISA. MMP-3 activity was assayed by casein zymography. RESULTS: All human TM cell strains produced detectable basal amounts of proMMPs and TIMPs. 12-O tetradecanoyl-phorbol-13-acetate was effective in increasing the levels of proMMP 1, -3, and -9 and TIMP-1. Its effect on proMMP-1 was concentration-dependent with an EC(50) of 2 to 3 nM. Interleukin (IL)-1alpha did not affect levels of proMMP-1 and -2 or the TIMPs, but was most efficacious in increasing proMMP-3 production with an EC(50) of 0.5 ng/mL. The IL-1alpha-induced upregulation of proMMP-3 correlated with an increase in MMP-3 activity. Tumor necrosis factor-alpha activated proMMP-3 production in some but not all cell strains. Platelet-derived growth factor-BB was generally ineffective in modulating MMP and TIMP levels. Prostaglandins E(2) and F(2alpha) at 10 micro M did not affect levels of proMMP-1 or -3. CONCLUSIONS: The expression of the different MMPs and TIMPs in human TM cells was independently regulated. Production of MMP-3 was maximally activated by IL-1alpha. The IL-1alpha-stimulated expression of MMP-3 provides a probable mechanism for IL-1alpha-enhanced aqueous outflow. PMID- 12882799 TI - Involvement of AP-1 in interleukin-1alpha-stimulated MMP-3 expression in human trabecular meshwork cells. AB - PURPOSE: Stromelysin-1 (MMP-3) degrades extracellular matrix and increases aqueous outflow. In the trabecular meshwork (TM), interleukin (IL)-1alpha is a potent inducer of MMP-3 expression. In different cells, IL-1alpha activates different signaling pathways, such as nuclear factor (NF)-kappaB-mediated protein expression, the phospholipase A(2) (PLA(2))-activated arachidonate cascade, and activator protein (AP)-1-associated transcription. In the present study, pharmacological tools were used to delineate the signaling mechanism involved in the effect of IL-1alpha on MMP-3 production in human TM cells compared with other ocular cells. METHODS: Human TM and three other ocular cells (ciliary muscle, corneoscleral fibroblast, and lamina cribrosa) were cultured in 24-well plates in the presence or absence of IL-1alpha, with or without specific inhibitors of selected signaling pathways. Secreted proMMP-3 was quantified by ELISA, and MMP-3 activity was assayed by casein zymography. RESULTS: IL-1alpha (5 ng/mL) increased proMMP-3 levels in human TM cells to 10- to 38-fold of control (P < 0.001). The effect of IL-1alpha was blocked by Go6976, a protein kinase C micro (PKC micro ) inhibitor; PD98059, a mitogen-activated protein kinase kinase (MEK) inhibitor; SB202190, a p38 inhibitor; and SR11302, an AP-1 inhibitor; but not by inhibitors of casein kinase II, NFkappaB, PLA(2), phospholipase D (PLD), cyclooxygenases, lipoxygenase, or sphingomyelinase. SR11302 did not inhibit the effect of IL 1alpha on MMP-3 production in the other ocular cells tested. CONCLUSIONS: Based on the pharmacological effects of the inhibitors, the data indicate that activation of PKC micro, MEK, and p38 leading to the activation of AP-1 is critical to the IL-1alpha-stimulated upregulation of MMP-3 in human TM cells. Therefore, it is likely that compounds that activate the AP-1 pathway would upregulate the production of MMP-3 and improve aqueous outflow. PMID- 12882800 TI - Aqueous outflow-enhancing effect of tert-butylhydroquinone: involvement of AP-1 activation and MMP-3 expression. AB - PURPOSE: To test the effect of stimulators of activator protein (AP)-1, on expression of stromelysin (MMP-3) in human TM cells and on aqueous outflow in perfused human anterior segments. METHODS: Change in MMP-3 expression was determined by immunoassay of proMMP-3 levels in the media of cultured human TM cells. Anterior segments of human donor eyes with or without glaucoma were perfused with vehicle or the AP-1 stimulator tert-butylhydroquinone (tBHQ). The outflow rates or intraocular pressure (IOP), and proMMP-3 levels in the perfusate were monitored. RESULTS: AP-1 stimulators, such as beta-naphthoflavone, 3 methylcholanthrene, and tBHQ, significantly upregulated (2-4-fold) TM cell expression of MMP-3. The stimulatory effect of tBHQ was concentration dependent, with an EC(50) of approximately 3 micro M, and was blocked by concomitant treatment with 100 nM SR11302, which sequesters AP-1. When nonglaucomatous human eyes were perfused with tBHQ (10 micro M), both outflow rates and perfusate proMMP-3 level increased significantly within the first 24 hours. The outflow effect of tBHQ was suppressed when SR11302 (100 nM) was added in the perfusate. tBHQ also lowered the IOP by more than 40% in perfused glaucomatous eyes. CONCLUSIONS: An AP-1 activator, tBHQ, upregulated expression of MMP-3 in cultured human TM cells and perfused human eyes and enhanced outflow ex vivo. These effects were blocked by sequestering AP-1, suggesting that activation of AP-1 can lead to increased MMP-3 production in the TM, which in turn improves outflow facility. This unique mechanism may provide a novel therapy for glaucoma. PMID- 12882801 TI - Expression of the inwardly rectifying K+ channel Kir2.1 in native bovine corneal endothelial cells. AB - PURPOSE: To determine the presence of Kir2.1 channels in native bovine corneal endothelial (BCE) cells and assess their contribution to the resting membrane potential. METHODS: RT-PCR and Western blot analysis were used to detect the expression of Kir2.1 mRNA and protein in native BCE cells. Whole-cell patch-clamp recording was used to characterize Kir2.1 currents in freshly isolated, single BCE cells, as well as in BCE cell clusters. The contribution of Kir2.1 channels to the membrane potential (V(m)) was assessed by whole-cell recording in the zero current clamp mode in the absence and presence of Ba(2+). RESULTS: RT-PCR analysis confirmed that Kir2.1 was expressed in the native BCE cells. Western blot analysis with native BCE cell protein and a polyclonal anti-Kir2.1 antibody revealed a approximately 60-kDa band that was blocked by the corresponding synthetic Kir2.1 peptide. Both single BCE cells and BCE cell clusters exhibited an inwardly rectifying K(+) (Kir) current that was dependent on the extracellular K(+) concentration. The Kir current was blocked by external Ba(2+) or Cs(+) in a voltage- and concentration-dependent manner. In 5 mM K(+) Ringer's, the V(m) of cell clusters averaged -40.0 +/- 4.1 mV (n = 14) and in 140 mM K(+) Ringer's it depolarized to -7.4 +/- 1.8 mV. Application of Ba(2+) in 5 mM K(+) Ringer's produced a concentration-dependent depolarization of V(m), with 10 mM Ba(2+) depolarizing V(m) from -53.4 +/- 4.8 mV to -27.8 +/- 6.3 mV (n = 6). CONCLUSIONS: Native BCE cells express functional Kir2.1 channels that help determine the membrane potential and probably also play a role in transendothelial transport. PMID- 12882802 TI - Examination of murine tear film. AB - PURPOSE: To define spatially any free aqueous layer in murine tear film. METHOD: A pre-zeroed microelectrode was touched to the superficial corneal epithelium and then raised in steps of 1 micro m through the murine tear film into the air and then retraced along the same path. Other murine tear films were partially probed with a spatial resolution of 0.1 micro m. The reference microelectrode was placed in a fragment of 3% polyacrylamide gel equilibrated against 154 mM NaCl and located on the nasal quadrant of the scleral conjunctiva. Other murine corneas were quick frozen in melting isopentane and freeze substituted or pretreated with cetylpyridinium chloride and then examined by transmission electron microscopy. RESULTS: The recorded electrical profiles of the tear film were reproducible in each preparation and showed a relatively uniform positive electrical potential throughout their whole thickness, except within 0.5 micro m of the epithelial surface when the potential reversed to negative values. The thickness of mouse tear film averaged 7.4 +/- 0.8 micro m (mean +/- SD, n = 40). The electron microscope images showed the murine tear film to have a relatively uniform positive electron density throughout the thickness. CONCLUSIONS: Electrical profiles of the murine tear film presented no evidence of a separate free aqueous phase. The tear film is observed as an aqueous gel that includes anion-exchanging polyelectrolytes throughout most of its thickness, but within 0.5 micro m of the epithelial surface, it changes to cation-exchanging polyelectrolytes. Electron microscope images provide some supporting evidence. PMID- 12882803 TI - Transplantation of syngeneic Schwann cells to the retina of the rhodopsin knockout (rho(-/-)) mouse. AB - PURPOSE: To determine whether subretinal Schwann cell transplantation can prolong the survival of photoreceptors in the rhodopsin knockout (rho(-/-)) mouse. METHODS: Schwann cells were prepared from postnatal day (PN) 5 to 7 mouse pups and grafted subretinally into the eyes of PN35 rho(-/-) mice. RT-PCR was performed on similarly prepared cells to determine growth factor production in vitro. Eyes were retrieved at PN70 for anatomic and statistical analysis. Control animals received grafts of fibroblasts or sham surgery. RESULTS: RT-PCR demonstrated the presence of message for ciliary neurotrophic factor (CNTF), brain-derived neurotrophic factor (BDNF), and glia-derived neurotrophic factor (GDNF) in the cultured Schwann cells. Schwann cell grafts produced a statistically significant rescue of photoreceptors in a restricted area of retina at PN70, but the effect was lost by PN140. Preserved inner segments could be identified, but outer segments were never present. Sham surgery also resulted in photoreceptor rescue but at a reduced level. Fibroblast grafts appeared to produce little or no rescue effect. Grafts of Schwann cells or fibroblasts and sham surgery induced a reactive Muller glial response. CONCLUSIONS: Schwann cells can prolong photoreceptor survival in the rhodopsin knockout mouse until at least PN70. PMID- 12882804 TI - Thresholds for activation of rabbit retinal ganglion cells with an ultrafine, extracellular microelectrode. AB - PURPOSE: To determine electrical thresholds required for extracellular activation of retinal ganglion cells as part of a project to develop an epiretinal prosthesis. METHODS: Retinal ganglion cells were recorded extracellularly in retinas isolated from adult New Zealand White rabbits. Electrical current pulses of 100- micro s duration were delivered to the inner surface of the retina from a 5- micro m long electrode. In about half of the cells, the point of lowest threshold was found by searching with anodal current pulses; in the other cells, cathodal current pulses were used. RESULTS: Threshold measurements were obtained near the cell bodies of 20 ganglion cells and near the axons of 19 ganglion cells. Both cathodal and anodal stimuli evoked a neural response in the ganglion cells that consisted of a single action potential of near-constant latency that persisted when retinal synaptic transmission was blocked with cadmium chloride. For cell bodies, but not axons, thresholds for both cathodal and anodal stimulation were dependent on the search method used to find the point of lowest threshold. With search and stimulation of matching polarity, cathodal stimuli evoked a ganglion cell response at lower currents (approximately one seventh to one tenth axonal threshold) than did anodal stimuli for both cell bodies and axons. With cathodal search and stimulation, cell body median thresholds were somewhat lower (approximately one half) than the axonal median thresholds. With anodal search and stimulation, cell body median thresholds were approximately the same as axonal median thresholds. CONCLUSIONS: The results suggest that cathodal stimulation should produce lower thresholds, more localized stimulation, and somewhat better selectivity for cell bodies over axons than would anodal stimulation. PMID- 12882805 TI - Pattern ERG correlates of abnormal fundus autofluorescence in patients with retinitis pigmentosa and normal visual acuity. AB - PURPOSE: To examine the functional significance of central abnormalities present in fundus autofluorescence (AF) images in patients with rod-cone dystrophy and good visual acuity. METHODS: Thirty patients were selected according to three criteria: a clinical diagnosis of retinitis pigmentosa (RP) confirmed with International Society for Clinical Electrophysiology of Vision (ISCEV) standard ERGs, a parafoveal ring of increased high density on fundus AF imaging, and a visual acuity of 20/30 or better. Macular function was assessed with pattern electroretinography (PERG) to checkerboard stimuli of different field sizes. Fundus AF imaging was performed with a confocal scanning laser ophthalmoscope. RESULTS: The radius of the parafoveal ring of high density varied between 1.5 degrees and 9 degrees. The PERG P50 amplitude correlated highly with the radius of the ring of increased autofluorescence (r = 0.80, P < 0.0005, n = 30). PERGs to smaller circular field sizes were present, but increasing field size to beyond that of the high-density autofluorescence ring did not produce further increases in P50 amplitude. There was a high correlation between the minimum stimulus size required to elicit a maximum-amplitude PERG and the radius of the ring (r = 0.87). CONCLUSIONS: The high correlation between AF imaging and PERG, an established technique in the assessment of central retinal function, demonstrates the likelihood that autofluorescence abnormalities have functional significance and may therefore be a valuable additional parameter in the monitoring of these patients. PMID- 12882806 TI - Specificity of subnormal deltaPO2 for retinal neovascularization in experimental retinopathy of prematurity. AB - PURPOSE: To test the hypothesis that in an experimental model of retinopathy of prematurity (ROP) rat pups that are at risk for but do not have retinal neovascularization (NV) exhibit a normal oxygenation response to a hyperoxic inhalation challenge. METHODS: Newborn Sprague-Dawley rats were raised under two varied oxygen conditions (50/10 or 40/15, indicating percent of oxygen in the air on alternate days) for 14 days and then allowed to recover in room air. Functional magnetic resonance imaging was used to determine the retinal oxygenation response (increase in partial oxygen pressure in the vitreous over the room air value, or deltaPO(2), in mm Hg) to hyperoxic inhalation challenge. Adenosine diphosphatase (ADPase)-stained retinas were analyzed to determine the NV incidence and severity. RESULTS: On postnatal day (P)20, the 40/15 procedure produced significantly (P < 0.05) lower incidence of NV than the 50/10 protocol (8% vs. 99%, respectively). Retinal deltaPO(2) during carbogen breathing of the 40/15 animals that did not have evidence of NV was not different (P > 0.05) from that of normal age-matched animals; later time points were not examined. At P26 and P34, in 50/10 rats that no longer had NV, retinal deltaPO(2)s during carbogen breathing were significantly (P < 0.05) lower than that of age-matched control pups. At P34 in control rats, but not in 50/10 rats, deltaPO(2) was 61% greater (P < 0.05) during carbogen breathing than during oxygen breathing. CONCLUSIONS: The results from 40/15 experiments, together with the authors' previous data in 50/10 rats, which documented subnormal retinal deltaPO(2) before and during NV, provides additional support that subnormal retinal oxygenation to an inhalation challenge is an important event associated with the development of NV. In addition, 50/10 rats that no longer demonstrated NV had a persistent subnormal retinal deltaPO(2), suggesting a continuing risk of development of additional retinal complications after resolution of the NV in ROP. PMID- 12882807 TI - Alkylphosphocholines inhibit proliferation of human retinal pigment epithelial cells. AB - PURPOSE: To investigate the effect and mechanism of action of alkylphosphocholines (APCs) on proliferation of human retinal pigment epithelium (RPE) cells and RPE-mediated collagen matrix contraction in vitro. METHODS: Cultured RPE cells of five human donors were treated with four APCs in the presence of fetal calf serum. Proliferation was assessed by the tetrazolium dye reduction (MTT) assay and by counting the number of cells dividing in culture. The effect of APCs on RPE-mediated matrix contraction was determined in three dimensional collagen gels. Cell viability was tested by the trypan blue exclusion assay. As a possible mechanism of APC action, protein kinase C (PKC) activity was quantified by scintillation counting of (32)P-labeled phosphate transferred to a PKC-specific substrate. RESULTS: All APCs inhibited RPE proliferation and RPE mediated collagen matrix contraction in a dose-dependent manner in vitro. The antiproliferative and anticontractile effect of APCs increased with elongation of the fatty acid chain beyond C20. IC(50)s of all APCs varied between 8.5 micro M (erucyl-phosphocholine, C22:1-PC), 9.0 micro M (Z)-12-heneicosenyl phosphocholine, C21:1-PC), 11.0 micro M (Z)-10-eicosenyl-phosphocholine, C20:1 PC), and 26.5 micro M (oleyl-phosphocholine, C18:1-PC). Trypan blue staining revealed a toxicity below 5% for all APCs within the concentration interval tested. PKC activity was significantly reduced by all four APCs, with C22:1-PC being the most effective. CONCLUSIONS: APCs inhibit proliferation of RPE cells and RPE-mediated matrix contraction in vitro at nontoxic concentrations. This effect seems to be exerted through inhibition of PKC activity. Therefore, APCs are promising candidates for treatment of RPE-mediated proliferative processes such as proliferative vitreoretinopathy. PMID- 12882808 TI - Ocular drug delivery targeting the retina and retinal pigment epithelium using polylactide nanoparticles. AB - PURPOSE: To study the kinetics of polylactide (PLA) nanoparticle (NP) localization within the intraocular tissues and to evaluate their potential to release encapsulated material. METHODS: A single intravitreous injection (5 micro L) of an NP suspension (2.2 mg/mL) encapsulating either Rh-6G (Rh) or Nile red (Nr) was performed. Animals were killed at various times, and the NPs localization within the intraocular tissues was studied by environmental scanning electron microscopy (ESEM), confocal microscopy, light microscopy histology, fluorescence microscopy, and immunohistochemistry. Eyes injected with blank NPs, free Rh, or PBS solution were used as the control. RESULTS: ESEM showed the flow of the NPs from the site of injection into the vitreous cavity and their rapid settling on the internal limiting membrane. Histology demonstrated the anatomic integrity of the injected eyes and showed no toxic effects. A mild inflammatory cell infiltrate was observed in the ciliary body 6 hours after the injection and in the posterior vitreous and retina at 18 to 24 hours. The intensity of inflammation decreased markedly by 48 hours. Confocal and fluorescence microscopy and immunohistochemistry showed that a transretinal movement of the NPs was gradually taking place with a later localization in the RPE cells. Rh encapsulated within the injected NPs diffused and stained the retina and RPE cells. PLA NPs were still present within the RPE cells 4 months after a single intravitreous injection. CONCLUSIONS: Intravitreous injection of PLA NPs appears to result in transretinal movement, with a preferential localization in the RPE cells. Encapsulated Rh diffuses from the NPs and stains the neuroretina and the RPE cells. The findings support the idea that specific targeting of these tissues is feasible. Furthermore, the presence of the NPs within the RPE cells 4 months after a single injection shows that a steady and continuous delivery of drugs can be achieved. PMID- 12882809 TI - Late-onset autosomal dominant macular dystrophy with choroidal neovascularization and nonexudative maculopathy associated with mutation in the RDS gene. AB - PURPOSE: To examine the molecular genetic basis and phenotypic characteristics of an unusual late-onset autosomal dominant macular dystrophy with features of age related macular degeneration (AMD) in a large family (SUNY901), by using linkage and mutation analyses. METHODS: Blood samples were collected from 17 affected members, 17 clinically unaffected members, and 5 unrelated spouses. Clinical analyses included a review of medical history and standard ophthalmic examination with fundus photography, fluorescein angiography, and electroretinography. Linkage and haplotype analyses were performed with microsatellite markers. Mutation analysis was performed by amplification of exons followed by sequencing. RESULTS: A wide spectrum of clinical phenotypes including exudative and nonexudative maculopathy was observed, with onset in the late fifth decade. Linkage analysis excluded most of the previously known maculopathy loci. Markers D6S1604 (Z(max) of 3.18 at theta = 0), and D6S282 (Z(max) of 3.18 at theta = 0) gave significant positive LOD scores and haplotype analysis localized the disease gene to a 9-centimorgan (cM) interval between markers D6S1616 and D6S459. Mutation analysis excluded the GUCA1A and GUCA1B genes and revealed a missense mutation in the RDS/peripherin gene leading to a Tyr141Cys substitution. A phenotype and haplotype comparison between this and a separate family with the Tyr141Cys mutation suggested the presence of a common ancestral haplotype. CONCLUSIONS: The RDS mutation in codon 141 is associated with an unusual AMD-like late-onset maculopathy. An apparent selective bias was noted favoring the transmission of the mutant allele. These observations broaden the spectrum of phenotypes associated with RDS gene mutations. PMID- 12882810 TI - Macrophage depletion inhibits experimental choroidal neovascularization. AB - OBJECTIVE: To investigate the role of macrophages in the development of laser induced choroidal neovascularization (CNV) by selective depletion with liposomal clodronate (Cl(2)MDP-LIP). METHODS: Laser photocoagulation was used to induce CNV in wild-type C57BL/6J mice. Animals were treated with intravenous (IV) and/or subconjunctival (SC) Cl(2)MDP-LIP or PBS-LIP at the following time points: 2 days before, immediately after, 2 days before and immediately after, or 2 days after laser injury. CNV responses were compared on the basis of en masse volumetric measurements and fluorescein angiography after laser photocoagulation. Macrophages were identified by immunostaining for F4/80, and vascular endothelial growth factor (VEGF) expression was quantified by ELISA. RESULTS: Macrophages invaded the site of laser injury within 1 day of photocoagulation and peaked at 3 days. IV Cl(2)MDP-LIP significantly decreased the volume of CNV and angiographic leakage when administered 2 days before and/or immediately after laser injury, but not when administered 2 days after injury. SC Cl(2)MDP-LIP significantly decreased lesion volume when coadministered with IV PBS-LIP but not IV Cl(2)MDP LIP. IV Cl(2)MDP-LIP was significantly more beneficial when administered 2 days before laser injury than immediately after, but combining SC Cl(2)MDP-LIP with IV treatment eliminated this difference. Reduction in CNV volume correlated with VEGF protein levels and number of infiltrating macrophages. CONCLUSIONS: Generalized macrophage depletion reduced the size and leakage of laser-induced CNV and was associated with decreased macrophage infiltration and VEGF protein. These findings define the role of the macrophage as a critical component in initiating the laser-induced CNV response. PMID- 12882811 TI - Macrophage depletion diminishes lesion size and severity in experimental choroidal neovascularization. AB - PURPOSE: Macrophage recruitment to the choroid has been proposed to contribute to the pathogenesis of choroidal neovascularization (CNV) in AMD. The study was conducted to determine whether treatment with clodronate liposomes (CL(2)MDP lip), which cause depletion of blood monocytes and lymph node macrophages, diminishes the severity of neovascularization in a mouse model of laser-induced CNV. METHODS: Laser-induced CNV was performed in female 16-month-old C57BL/6 mice. Macrophages were depleted by use of CL(2)MDP-lip intraperitoneally and subcutaneously 72 and 24 hours before and every 2 to 3 days after laser injury. Control mice received injections of either PBS alone or PBS liposomes. Blood monocyte and choroidal macrophage depletion were documented by flow cytometry and choroidal flatmount preparation analysis, respectively. Two weeks after laser injury, mice were injected intravenously with fluoresceinated dextran. The right eyes were removed and prepared for flatmount analysis of CNV surface area (in relative disc areas or DA), vascularity (relative fluorescence), and cellularity (propidium iodide stain). The mice were then perfused with 10% formaldehyde, and the left eyes were removed for histopathology. The means of the various parameters for four CNV lesions per eye were calculated. Fluorescein angiography was also performed. RESULTS: Flow cytometry of circulating monocytes and immunohistochemical analysis of choroidal macrophage density confirmed the effective depletion of blood monocytes and choroidal macrophages respectively in CL(2)MDP-lip-treated mice. Compared with the control, flatmount analysis of macrophage depleted mice demonstrated a significant reduction in size of the CNV area (2.8 +/- 0.5 DA vs. 1.4 +/- 0.1 DA; P < 0.043). The treated group also revealed less vascularity (1.6 +/- 0.1 units vs. 1.1 +/- 0.0 units; P < 0.0092) and cellularity of CNV lesions (3.3 +/- 0.6 DA vs. 1.7 +/- 0.1 DA, P < 0.04). Histopathology revealed that, in the macrophage-depleted group, CNV was smaller in diameter (1270 +/- 73 pixels vs. 770 +/- 82 pixels, P < 0.0006) and thickness (120 +/- 7 pixels vs. 96 +/- 7 pixels, P < 0.019). CONCLUSIONS: Macrophage depletion using CL(2)MDP-lip reduces size, cellularity, and vascularity of CNV. This observation supports the hypothesis that macrophages contribute to the severity of CNV lesions. PMID- 12882812 TI - De novo mutation in the RP1 gene (Arg677ter) associated with retinitis pigmentosa. AB - PURPOSE: The Arg677ter mutation in the RP1 gene is one of the most common causes of autosomal dominant retinitis pigmentosa (RP). In the current study, a de novo Arg677ter RP1 gene mutation was identified in a patient with RP. METHODS: RP1 gene mutation screening was performed in probands with simplex RP. In one proband with the RP1 mutation, paternity was established by analyzing 24 short tandem repeat polymorphisms. Additional candidate RP genes, including rhodopsin, RDS/peripherin, RP2, and RPGR, were also examined in this proband. Phenotype was characterized with psychophysics, electroretinography, and optical coherence tomography. RESULTS: An RP1 (Arg677ter) mutation was identified in one of the patients with simplex RP, but the sequence change was not detected in his parents. Parentage was confirmed, and other candidate genes were negative for mutations. Retinal function and cross-sectional imaging studies in the patient indicated greater rod than cone dysfunction with a photoreceptor basis for the abnormalities. CONCLUSIONS: The de novo origin of an RP1 (Arg677ter) mutation in a patient with simplex RP suggests that this common autosomal dominant RP mutation can arise independently in the population and supports the hypothesis of a mutational hotspot in the RP1 gene. PMID- 12882813 TI - Proapoptotic bcl-2 family members, Bax and Bak, are essential for developmental photoreceptor apoptosis. AB - PURPOSE: Apoptosis has been implicated in retinal development and degeneration, but the specific apoptotic pathways used are incompletely understood. The purpose of this study was to characterize the roles in retinal development of the proapoptotic Bcl-2 family members Bax and Bak. METHODS: Eyes from mice at postnatal day (P)7, during the peak of developmental apoptosis in the retina, were processed for TdT-dUTP terminal nick-end labeling (TUNEL) to determine whether Bax knockout or double Bax/Bak knockout causes a defect in developmental apoptosis. Adult (>2-month-old) eyes from wild-type, Bak(-/-), Bax(-/-), and Bax( /-)Bak(-/-) mice were analyzed by histology and immunocytochemistry to identify persistent retinal cells. RESULTS: Adult Bax(-/-)Bak(-/-) eyes showed significant increases in the number of inner retinal cells, with an almost complete absence of TUNEL-positive cell death at P7. Some of these persistent cells in the inner retina notably included rod photoreceptors that normally undergo apoptosis after failure to migrate to the outer retina. These inner nuclear layer (INL) rods contained markers of early rod differentiation: rod opsin, arrestin, and recoverin. However, they did not form ectopic outer segments or contain the associated markers ROM-1, peripherin-2, and RP1. CONCLUSIONS: Bax and Bak are important for retinal development and are the first apoptotic factors identified as essential for developmental photoreceptor apoptosis. Future studies will investigate the potential role of Bax and Bak in mediating pathologic photoreceptor death. PMID- 12882814 TI - GFAP promoter drives Muller cell-specific expression in transgenic mice. AB - PURPOSE: In an attempt to identify Muller cell-specific promoters and to better understand the gene regulatory mechanisms in retinal glial cells, the expression of the glial fibrillary acidic protein (GFAP) gene was studied in Muller cell cultures and in GFAP-enhanced green fluorescent protein (EGFP) transgenic mice. METHODS: A transfection assay of GFAP-luciferase constructs carrying a series of nested deletions was performed in an established Muller cell line. For in vivo analysis, transgenic mice were generated by injecting a construct carrying a 2.5 kb, 5' fragment of the mouse GFAP gene linked to the EGFP gene. Isolated retinas from transgenic mice were screened for GFP expression. Subsequently, the identity of the GFP-expressing cells was established by immunostaining cryostat sections of the retina with antibodies against Muller cell antigenic markers. Induction of the transgene and the endogenous GFAP gene was examined by injecting ciliary neurotrophic factor (CNTF) into the eye. RESULTS: The DNA transfection data suggested that proximal 5' sequences of the GFAP gene are sufficient to direct high-level reporter expression in Muller cell cultures. In transgenic mice, GFP fluorescence appeared in radially oriented processes that spanned almost the entire thickness of the retina. Immunostaining with antibodies to cellular retinaldehyde-binding protein (CRALBP) and glutamine synthetase showed that the GFP-expressing cells were Muller cells. GFP-expressing Muller cells were observed in the retinas of both albino and pigmented transgenic mice. In eyes injected with CNTF, both GFP and GFAP levels were highly elevated. These observations suggest that the 2.5-kb, 5' GFAP sequence can direct inducible reporter gene expression in Muller cells. In addition to Muller cells, a few GFP-labeled astrocytes were present in the adult retina. In the developing retina, GFP expressing astrocytes were first present at the optic nerve head, and as development progressed, the cells gradually moved toward the periphery of the retina and acquired their adult, stellate morphology. CONCLUSIONS: The present study shows that the 2.5-kb, 5' flanking region of the mouse GFAP gene can be used to express GFP, and possibly other genes, specifically in Muller cells in the mouse retina. Furthermore, expression of the transgene can be upregulated by intravitreal injection of CNTF. PMID- 12882815 TI - Disruption of kinesin II function using a dominant negative-acting transgene in Xenopus laevis rods results in photoreceptor degeneration. AB - PURPOSE: Kinesin II is a motor protein that moves on microtubules and whose importance in ciliary and flagellar transport has been well documented. In the current study, the role of kinesin II in rod photoreceptors was examined by expressing a dominant negative-acting transgene that disrupts kinesin II function in Xenopus laevis rods of transgenic tadpoles. METHODS: A previously characterized dominant negative-acting kinesin II transgene tagged with enhanced green fluorescent protein (EGFP) driven by the Xenopus rod opsin promoter was used to make Xenopus transgenic tadpoles to disrupt kinesin II function specifically in rod photoreceptors. Transgenic tadpole retinas were examined to ascertain transgene expression pattern and morphologic phenotype. Rod-to-cone ratios were determined in experimental and control retinas. RESULTS: Visualized by its EGFP tag, the kinesin II transgene was expressed in rods in a mosaic pattern in the retina. Subcellular localization of transgenic kinesin II was similar to that of endogenous kinesin II subunit photoreceptor expression-that is, it was localized to the connecting cilium, inner segment, and synapse. However, in kinesin II transgene-expressing animals, fluorescence was transient. Ocular fluorescence was lost 6 days after its first detection. The disappearance of fluorescence was due to degeneration of rods expressing the transgene. Retinas of 7- to 9-day old kinesin II transgenic tadpoles had significantly fewer rods than did control retinas. CONCLUSIONS: The observation that rod degeneration is produced by expression of a dominant negative-acting kinesin II transgene in Xenopus rods is consistent with previous studies in mice, suggesting that kinesin II function is required for photoreceptor survival. PMID- 12882816 TI - Expression and localization of bestrophin during normal mouse development. AB - PURPOSE: Best macular dystrophy is caused by mutations in the VMD2 gene, which encodes the protein bestrophin. The purpose of this study was to determine the postnatal onset of expression of bestrophin mRNA and protein in the mouse retinal pigment epithelium (RPE). METHODS: Rabbit anti-mouse bestrophin polyclonal antisera designated Pab-003 was generated against a peptide derived from the C terminus of mouse bestrophin and characterized by Western blot and immunofluorescence staining of transfected cells. Expression of bestrophin mRNA during ocular development was studied with quantitative PCR. Bestrophin protein expression in the developing eye was observed by using immunohistochemistry. The onset of mouse phototransduction was determined by conventional electroretinography (ERG). RESULTS: Bestrophin mRNA was detected at embryonic day 15 in whole mouse eyes by RT-PCR. Real-time quantification of mouse bestrophin mRNA levels indicated that the highest levels of mRNA were present in the early postnatal period. In contrast, bestrophin in the RPE was first detected at postnatal day (P)10 by immunohistochemistry. Phototransduction, as determined by the presence of an ERG a-wave, was first observed at P10. CONCLUSIONS: The results of this study show that mouse bestrophin mRNA is present in the eye during embryogenesis and significantly precedes the onset of bestrophin protein expression at P10. The appearance of bestrophin in the basolateral plasma membrane of the RPE is coincident with the first detectable ERG a-wave. Because bestrophin is thought to play a role in generating the light peak, a late response of the ERG, these data support a temporal role for bestrophin in RPE responses to light. Furthermore, bestrophin protein appears to be a very late marker of RPE differentiation and to be subject to strong translational control. PMID- 12882817 TI - Comparative proteome analysis of native differentiated and cultured dedifferentiated human RPE cells. AB - PURPOSE: Dedifferentiation of retinal pigment epithelial (RPE) cells is a crucial event in the pathogenesis of proliferative vitreoretinopathy (PVR). This study was designed to improve the understanding of RPE cell dedifferentiation in vitro. The protein expression pattern of native differentiated RPE cells was compared with that of cultured, thereby dedifferentiated, RPE cells. METHODS: Differentiated native human RPE cells and monolayers of dedifferentiated cultured primary human RPE cells were processed for two-dimensional (2-D) electrophoresis. Total cellular proteins were separated by isoelectric focusing using immobilized pH gradients (IPG 3-10) and electrophoresis on 9% to 15% gradient polyacrylamide gels. Proteins were visualized by silver staining. Silver-stained gel spots were excised, digested in situ, and analyzed by matrix-assisted laser desorption ionization time of flight (MALDI-TOF) mass spectroscopy (MS). The resultant peptide mass fingerprints were searched against the public domain NCBInr, MSDB, and EnsemblC databases to identify the respective proteins. RESULTS: One hundred seventy nine protein spots were analyzed and classified into functional categories. Proteins associated with highly specialized functions of the RPE, which are required for interaction with photoreceptor cells, including RPE65, cellular retinaldehyde-binding protein (CRALBP), and cellular retinol-binding protein (CRBP), were absent in dedifferentiated cultured RPE cells, whereas proteins involved in phagocytosis and exocytosis, including cathepsin D and clathrin were still present. Dedifferentiated RPE cells displayed a strong shift toward increased expression of proteins associated with cell shape, cell adhesion, and stress fiber formation, including cytokeratin 19, gelsolin, and tropomyosins, and also acquired increased expression of factors involved in translation and tumorigenic signal transduction such as annexin I and translation initiation factor (eIF)-5A. CONCLUSIONS: Dedifferentiation of human RPE cells in vitro results in downregulation of proteins associated with highly specialized functions of the RPE and induces the differential expression of proteins related to cytoskeleton organization, cell shape, cell migration, and mediation of proliferative signal transduction. These in vitro data suggest that the dedifferentiated status of RPE cells per se may initiate PVR. Further investigation of candidate proteins may identify additional targets for treatment or prevention of diseases associated with RPE dedifferentiation. PMID- 12882818 TI - Cloning, mapping, and retinal expression of the canine ciliary neurotrophic factor receptor alpha (CNTFRalpha). AB - PURPOSE: To clone, map, and determine the site of expression (mRNA and protein) of the alpha subunit of the receptor for ciliary neurotrophic factor (CNTFRalpha) in the normal adult canine retina. METHODS: The complete coding sequence of the canine CNTFRalpha cDNA was cloned, and radiation hybrid (RH) mapping was used to determine the chromosomal localization of the gene. CNTFRalpha mRNA expression in retina and other tissues was examined by reverse transcription-polymerase chain reaction. The cellular distribution of CNTFRalpha in the canine retina was studied by in situ hybridization and immunocytochemistry. RESULTS: Canine CNTFRalpha shares a high degree of homology with the human, mouse, and rat coding sequences, both at the nucleotide and amino acid level, but has lower homology with the chicken. CNTFRalpha was RH mapped to CFA 11 (Canis familiaris autosome 11) in the dog, a region showing homology to the short arm of human chromosome 9 (9p13). The gene is transcribed in retina, brain, spleen, lung, liver, and kidney. In the retina, CNTFRalpha was highly expressed by photoreceptors, but both the transcript and protein were also found in the RPE, inner nuclear layer, and ganglion cells. CONCLUSIONS: These findings demonstrate that CNTFRalpha is expressed by rods and cones in the normal adult canine retina and suggest that ciliary neurotrophic factor (CNTF) could have a direct photoreceptor rescue effect by binding to CNTFRalpha in these cells. This could open novel pathways for the treatment of retinal degeneration in animal models and humans. PMID- 12882819 TI - Combretastatin A-4 phosphate suppresses development and induces regression of choroidal neovascularization. AB - PURPOSE: Combretastatin A-4 (CA-4) is a naturally occurring agent that binds tubulin and causes necrosis and shrinkage of tumors by damaging their blood vessels. In this study the effect of a CA-4 prodrug, combretastatin A-4-phosphate (CA-4-P), was tested in two models of ocular neovascularization. METHODS: The effect of CA-4-P was quantitatively assessed in transgenic mice with overexpression of vascular endothelial growth factor in the retina (rho/VEGF mice) and mice with choroidal neovascularization (CNV) due to laser-induced rupture of Bruch's membrane. RESULTS: In rho/VEGF mice, daily intraperitoneal injections of 4.0 mg/kg CA-4-P starting at postnatal day (P)7, the time of onset of transgene expression, resulted in a significant reduction in the number of neovascular lesions and total area of neovascularization per retina at P21, compared with vehicle-injected mice. In mice with laser-induced rupture of Bruch's membrane, daily intraperitoneal injections of 75 or 100 mg/kg CA-4-P resulted in a significant reduction in the area of CNV at rupture sites compared with vehicle-injected mice. In mice with established CNV, daily intraperitoneal injections of 100 mg/kg CA-4-P for 1 week resulted in a significant reduction in CNV area at rupture sites compared with the baseline area before treatment or the area of CNV in vehicle-treated mice. CONCLUSIONS: These data indicate that CA-4-P suppresses the development of VEGF-induced neovascularization in the retina and both blocks development and promotes regression of CNV. Therefore, CA-4-P shows potential for both prevention and treatment of ocular neovascularization. PMID- 12882820 TI - The kinase inhibitor PKC412 suppresses epiretinal membrane formation and retinal detachment in mice with proliferative retinopathies. AB - PURPOSE: Platelet-derived growth factor (PDGF) is an important stimulatory factor for proliferative retinopathies. Expression of PDGF-B in the retinas of transgenic mice (hemizygous rho/PDGF-B mice) results in rapid-onset retinal detachment caused by proliferation of glial cells, endothelial cells, and pericytes, whereas expression of PDGF-AA (homozygous rho/PDGF-A or PDGF-AA mice) causes slowly progressive retinal detachment from proliferation of glial cells. In this study, we investigated the effect in rho/PDGF-B and rho/PDGF-AA mice of several different receptor kinase inhibitors. METHODS: Hemizygous rhoPDGF-B or homozygous rho/PDGF-A mice were treated orally with PKC412 (an inhibitor of PDGF, VEGF, and c-kit receptor kinases and several isoforms of PKC), PTK787 (an inhibitor of PDGF, VEGF, and c-kit receptor kinases), SU1498 (an inhibitor of VEGF receptor kinases), imatinib mesylate (an inhibitor of PDGF, c-kit, and v-abl receptor kinases), or vehicle, and at appropriate time points epiretinal membrane (ERM) formation and retinal detachment were quantified. RESULTS: In either rho/PDGF-B or rho/PDGF-A mice, oral administration of PKC412 or PTK787, but not SU1498 or imatinib mesylate, significantly reduced ERM formation. PKC412 reduced the incidence of severe retinal detachments in both models and PTK787 did so in homozygous rho/PDGF-A mice. CONCLUSIONS: These data indicate that PKC412 (and possibly PTK787) has appropriate activity and sufficient intraocular bioavailability after oral administration to prevent retinal detachment in models of proliferative retinopathy. PKC412 should be considered for treatment of vascular and nonvascular proliferative retinopathies in humans. PMID- 12882821 TI - Proteins modified by malondialdehyde, 4-hydroxynonenal, or advanced glycation end products in lipofuscin of human retinal pigment epithelium. AB - PURPOSE: Lipofuscin (LF) accumulation in the retinal pigment epithelium (RPE) is associated with age and various retinal diseases. Toxic LF compounds may interfere with normal RPE function. Oxidative modification of proteins was determined in LF granules from human eyes. METHODS: LF was isolated from the RPE choroid complex of 10 pairs of donor eyes by gradient ultracentrifugation. Protein compounds were separated by two-dimensional (2-D) gel electrophoresis and screened by Western blot analysis for lipid peroxidation- or glucoxidation induced damage-in particular, by malondialdehyde (MDA), 4-hydroxynonenal (HNE), and advanced glycation end products (AGEs). Identity of the immunostained proteins was revealed using 2-D software for comparison of the spot position with Coomassie-stained 2-D gels of the same samples. RESULTS: By comparing the results taken from the authors' previous proteome analysis of RPE LF with an immunoblot analysis of the same samples, this study shows that a variety of LF-associated proteins were damaged by aberrant covalent modifications of MDA, 4-HNE, and AGEs. Several proteins were altered by two or three different modification types. Modified mitochondrial proteins indicated that autophagy of altered proteins also contributed to lipofuscin formation. CONCLUSIONS: The identification of lipid peroxidation and glucoxidation products in proteinaceous LF components in human RPE supports the hypothesis that these compounds are involved in lipofuscinogenesis and may contribute to the cytotoxic effects of LF in retinal diseases such as age-related macular degeneration and Stargardt disease. Their identification may help to identify potential future treatment targets. PMID- 12882822 TI - Novel role for a complement regulatory protein (CD46) in retinal pigment epithelial adhesion. AB - PURPOSE: There is increasing evidence that the complement system may play a significant role in one of the leading diseases causing blindness in the elderly population, age-related macular degeneration. In this study, a novel role in the retina for a regulatory protein in the complement system, CD46, is proposed. METHODS: The retinal pigment epithelium (RPE) was obtained from human donor eyes as well as human immortalized RPE cell lines (ARPE19). Immunohistochemistry and confocal microscopy were used to immunolocalize CD46 and beta1 integrin. Immunoprecipitation experiments with antibodies to either CD46 or beta1 integrin were performed on RPE cell lysates. A cell adhesion assay was used to determine the proportion of RPE cells that adhere to Bruch's membrane explants from donor eyes. RESULTS: Immunohistochemistry and confocal microscopy demonstrated that CD46 was polarized to the basal surface of the RPE along with beta1 integrin, shown previously to be involved in RPE adhesion. Immunoprecipitation experiments demonstrated that CD46 and beta1 integrin coprecipitated from RPE cell lysates when either protein was used as the precipitating antibody. The adhesion assay showed that antibodies to either CD46 or beta1 integrin reduced RPE adhesion to the surface of Bruch's membrane compared with the control. CONCLUSIONS: These findings suggest that this complement regulatory protein, which protects host cells from autologous complement attack, may have a functional interaction with beta1 integrin in the eye that is related to RPE adhesion to its basement membrane and Bruch's membrane. PMID- 12882823 TI - Functional damage to inner and outer retinal cells in experimental glaucoma. AB - PURPOSE: To investigate the cellular sources underlying the functional damage observed by multifocal electroretinography (mfERG) responses of glaucomatous eyes of monkeys. METHODS: First- and second-order (K1 and K2, respectively) mfERG responses of three normal and three experimentally induced glaucomatous eyes of cynomolgus monkeys were measured at two different levels of luminance. Retinal contributors to the responses were isolated by intravitreal injections of pharmacological agents that suppress specific retinal cells. gamma-Aminobutyric acid (GABA) and glycine were administered to block inner retinal function, followed by 2-amino-4-phosphonobutyric acid (APB), to block ON-bipolar cells. RESULTS: An inner retinal component removed by GABA and glycine was found in both the normal and glaucomatous eyes. However, it was attenuated in the latter, correlating with changes observed in the baseline K1 responses. Delays in the latency of outer retinal components were found in the responses of the glaucomatous eyes. K2 responses were dominated by an inner retinal contribution and were diminished in the responses of glaucomatous eyes. The outer retina responded to increased luminance with a shorter implicit time. A distinct wave part of the inner retinal component responded to increased luminance with increased amplitudes. CONCLUSIONS: The integration of the retinal sources forming the mfERG response was compared between normal and glaucomatous monkey eyes. Both inner and outer retinal functions were aberrant in the responses of the glaucomatous eyes, with the attenuation of the inner retinal function more conspicuous. Nevertheless, glaucomatous eyes retained certain inner retinal activity, despite the advanced stage of disease. K2 responses were more sensitive to glaucomatous changes than were K1 responses. PMID- 12882824 TI - Scotopic electroretinogram in term infants born of mothers supplemented with docosahexaenoic acid during pregnancy. AB - PURPOSE: To test the hypothesis that the supplementation of the diets of pregnant women with a fish oil rich in docosahexaenoic acid (DHA) enhances retinal development in their healthy term infants, as measured during the early postnatal period by the electroretinogram (ERG). METHODS: One hundred pregnant women were randomized to receive either a fish oil (n = 50) or a placebo oleic acid dietary supplement (n = 50) from 15 weeks of pregnancy until delivery. Total fatty acids in red blood cells (RBCs) and plasma were measured in mothers at 15 and 28 weeks of pregnancy and at delivery and in their infants in umbilical cord blood. Infant retinal development was assessed within the first week of life with full-field ERGs that included a scotopic blue intensity series (n = 41) and a bright white flash (2.0 log cd-s/m(2); n = 44). RESULTS: Infants born of mothers who received supplements did not differ at birth in weight, gestational age, or any other standard variable. Infant DHA status at birth, as measured from umbilical cord blood, did not differ significantly between maternal supplementation groups. ERG implicit times, amplitudes, and parameters of the stimulus-response function did not differ significantly between infants in the maternal supplemented and placebo groups. There was, however, a relationship between infant DHA status and maturity of the retina at birth, regardless of maternal supplementation group. A measure of retinal sensitivity (log sigma) correlated significantly (P < 0.005) with DHA status (as a percentage of total fatty acid; TFA) in infant cord blood. Infants in the highest quartile for cord blood DHA had higher retinal sensitivity compared with infants in the lowest quartile. Infants in the highest quartile for plasma DHA, both as a percentage of TFA and concentration, were born at a significantly later gestational age than were infants in the lower quartiles. CONCLUSIONS: These findings demonstrate an association between the DHA status of term infants and retinal sensitivity, suggesting an essential role of this long chain polyunsaturated fatty acid (LCPUFA) in the development and function of the retina. However, maternal DHA status was not significantly associated with infant retinal sensitivity and no direct effect of maternal supplementation was observed. PMID- 12882825 TI - The effects of constant and diurnal illumination of the pineal gland and the eyes on ocular growth in chicks. AB - PURPOSE: To investigate the effects of constant or 12-hour cyclic illumination of the pineal gland and the eyes on the growth of the chick eye. METHODS: Chicks (Gallus gallus, Cornell K Strain) were raised either under a 12-hour light-dark cycle of normal light or under constant light, with or without opaque removable hoods that covered the top of the head for 12 hours each day. A second group of chicks was raised under constant light with opaque eye covers that were worn on either both eyes or only the right eye for 12 hours each day. Chicks were placed in the experimental conditions on the third day after hatching and raised for 3 weeks. RESULTS: Pineal gland hoods and eye covers worn 12 hours a day significantly (P < 0.0001) protected the chicks from hyperopia under constant light conditions. They also reduced the flattening of the cornea caused by constant light. Most striking was the protection afforded the uncovered eye from constant light's effects by the periodic covering of the opposite eye. CONCLUSIONS: A diurnal light-dark rhythm presented to one of three photosensitive organs (the pineal gland and both eyes) can protect the eyes from the effects of constant light. This is most probably due to the maintenance of a melatonin rhythm in the organ receiving the diurnal light rhythm. PMID- 12882826 TI - The role of lenticular senescence in age-related color vision changes. AB - PURPOSE: It has been reported that greater age-related losses in sensitivity occur for short-wavelength visual stimuli than for medium- and long-wavelength visual stimuli. The purpose of the current experiment was to determine to what extent optical, receptoral, and postreceptoral factors contribute to these age related changes in color vision. METHODS: One hundred two observers (ages 18-87) completed a minimum motion task to determine isoluminance between red and green and between red and blue. A motion-nulling task was also performed to assess the L-M postreceptoral chromatic mechanism. RESULTS: No significant age-related changes occurred in red-green isoluminance values. Red-blue isoluminance values showed a significant and systematic decrease with age in observers with phakic eyes. Pseudophakic eyes in older subjects performed this task as well as phakic eyes in young subjects. The motion-nulling results demonstrated small age-related losses in the postreceptoral color mechanisms. CONCLUSIONS: The findings of this experiment, particularly those of the red-blue isoluminance task, indicate that the optical factor of lenticular senescence is the main contributor to the age related changes observed in color vision. A model based on age-related changes in lenticular absorbance shows good fit with the experimental data of observers with phakic eyes, suggesting that optical factors are the main cause of the age related changes in these color vision tasks. PMID- 12882827 TI - The cytolytic activity of natural killer cells is not involved in the restriction of Mycobacterium avium growth. AB - Severe combined immunodeficiency (SCID) mice were used to analyze the role of NK cells in resistance to Mycobacterium avium. The neutralization of IFN-gamma in these animals led to an exacerbation of the infection associated with a reduction in macrophage activation, suggesting a role for NK cells in innate immunity to mycobacteria. In contrast, administration of anti-asialo-GM(1) polyclonal serum or mAb specific for Thy1.2 did not affect mycobacterial growth or macrophage activation despite causing the almost complete abrogation of the natural cytolysis of a tumor cell target. Treatment with anti-asialo-GM(1)-specific serum depleted only two-thirds of the Thy1.2+ spleen cells, and anti-Thy1.2 treatment allowed for the persistence of a small number of cells still exhibiting an NK cell marker recognized by mAb DX5 and able to express IFN-gamma as analyzed by flow cytometry. In vivo treatment of B6.SCID mice with anti-NK1.1 mAb again failed to affect resistance to infection and allowed for the persistence of 2-8% of IFN-gamma-producing cells, many of them still expressing the DX5 marker. In vitro depletion studies showed that removal of IFN-gamma-expressing cells required the combined action of anti-Thy1.2, anti-Ly49C and DX5 antibodies in the presence of complement. Our data show that resistance to M. avium mediated by NK cells is independent of their cytolytic activity, and that there is a marked phenotypic and functional heterogeneity of the NK cell lineage in vivo during infection. PMID- 12882828 TI - Spontaneous production of anti-IFN-alpha and anti-IL-12 autoantibodies by thymoma cells from myasthenia gravis patients suggests autoimmunization in the tumor. AB - Myasthenia gravis (MG) is mediated by autoantibodies to the acetylcholine receptor (AChR), expressed in muscle and rare thymic myoid cells. Most early onset cases show thymic lymph node-type infiltrates, including pre-activated plasma cells spontaneously producing anti-AChR antibodies. Since these are not evident in the associated thymomas found in another 10% of MG patients, AChR specific B cells must be autosensitized elsewhere. Unexpectedly, at diagnosis, >70% of MG/thymoma patients also have high-titer neutralizing autoantibodies to IFN-alpha, and >50% to IL-12; moreover, titers increase strikingly if the thymomas recur, indicating a closer tumor relationship than for anti-AChR. To investigate this, we have measured autoantibody production by cells cultured from thymomas, any available thymic remnants and blood, with or without the B cell stimulant pokeweed mitogen (PWM). To check autoantibody specificity and clonal origins, we isolated Fabs from two combinatorial libraries from producer thymus/thymoma cells. Surprisingly, thymoma cells spontaneously produced antibodies to IFN-alpha and/or IL-12 in >40% of seropositive cases, showing typical plasma cell behavior, whereas they produced anti-AChR only after PWM stimulation. We isolated 15 combinatorial Fabs to IFN-alpha (versus only one to AChR). Their strong binding in radio-immunoprecipitation and Western blots implies high affinities. The four Fabs tested neutralized anti-viral actions of IFN-alpha. The diverse V genes clearly showed ongoing antigen-driven selection. These results imply pre-activation in situ by native IFN-alpha/IL-12 expressed within a 'dangerous' tumor microenvironment. With these molecules, it should be easier to identify provoking cell type(s) that may give novel additional clues to autoimmunization against T-cell epitopes from the more complex AChR. PMID- 12882829 TI - Immunization with a mycobacterial lipid vaccine improves pulmonary pathology in the guinea pig model of tuberculosis. AB - Lipids and glycolipid molecules derived from Mycobacterium tuberculosis can be presented to T cells by CD1 cell-surface molecules in humans. These lipid specific T cells are cytolytic, secrete pro-inflammatory cytokines and have bactericidal activity. Here, we describe studies in which lipids from M. tuberculosis were incorporated into liposomes with adjuvant and tested as vaccines in a guinea pig aerosol tuberculosis challenge model. Animals vaccinated with mycobacterial lipids showed reduced bacterial burdens in the lung and spleen at 4 weeks after infection. In addition, the lungs of lipid-vaccinated animals also had significantly less pathology, with granulomatous lesions being smaller and more lymphocytic. In contrast, animals receiving only vehicle control immunizations had granulomatous lesions that were larger and often contained caseous necrotic centers. Quantification of histopathology by morphometric analysis revealed that the overall percentage of lung occupied by diseased tissue was significantly smaller in lipid-vaccinated animals as compared to vehicle control animals. In addition, the mean area of individual granulomatous lesions was found to be significantly smaller in both lipid- and bacillus Calmette-Guerin vaccinated guinea pigs. These data support an important role for lipid antigens in the immune response to M. tuberculosis infection, potentially through the generation of CD1-restricted T cells. Immunogenic lipids thus represent a novel class of antigens that might be included to enhance the protective effects of subunit vaccine formulations. PMID- 12882830 TI - Differential recognition of a retinal autoantigen peptide and its variants by rat T cells in vitro and in vivo. AB - Previously we have described the role of two 14mer peptides in autoimmune uveitis, PDSAg from the retinal autoantigen S-antigen (S-Ag) and B27PD from the sequence of disease-associated HLA-B molecules, which show antigenic mimicry. The retinal peptide gave rise to severe uveitis in the Lewis rat model of experimental autoimmune uveitis (EAU) and was effective in inducing oral tolerance, while the HLA peptide B27PD caused only mild disease, but it was at least equally effective in preventing uveitis by oral tolerance. Here, we further defined the major T cell epitopes on both peptides responsible for the different functions. For this purpose we tested C- and N-terminal truncations, and chimeras consisting of amino acid sequences of both peptides in vitro and in vivo. We were able to determine the motif for binding to Lewis rat MHC class II as well as those amino acids important for recognition by T cells specific for the retinal peptide. The minimal MHC-binding nonamer peptide of PDSAg was not recognized by TCR, and we also found striking differences of T cell recognition in vitro and in vivo. The ability to induce oral tolerance was not closely correlated with uveitogenicity or with strong binding to MHC class II molecules. Our data furthermore demonstrate the importance of specific and exact trimming of peptides to be presented on MHC class II, suggesting that the presentation of cryptic epitopes is favored or prevented by existence of multiple MHC-binding motifs within a certain amino acid sequence, which can result in different or altered T cell reactions. PMID- 12882831 TI - T-bet regulates T-independent IgG2a class switching. AB - The IgG2a Ig subclass plays a critical role in the pathogenesis of humoral autoimmunity and protection against pathogens. The T-box transcription factor T bet has been implicated as a critical mediator of class-switch recombination (CSR) to IgG2a, but its relative importance to this process in various immune contexts remains incompletely defined. We report here that, surprisingly, T-bet is selectively required for IgG2a class switching in response to T-independent, but not T-dependent, stimuli. Specifically, T-dependent signaling through CD40, in contrast to T-independent signaling via lipopolysaccharide, can bypass a requirement for T-bet in IgG2a germline transcription and subsequent isotype switching. In contrast, T-bet-deficient B cells undergo class switching to other IgG isotypes at least as well as wild-type counterparts. Thus, T-bet is a class specific regulator of IgG CSR and represents a unique regulator of B cell differentiation by participating in a T-independent, but not a T-dependent, activation pathway. T-bet-deficient B cells therefore represent a novel paradigm by which to investigate the regulation of humoral immune responses. PMID- 12882832 TI - Deletion, but not anergy, is involved in TGF-beta-treated antigen-presenting cell induced tolerance. AB - Intravenous injection of transforming growth factor (TGF-)-beta-treated antigen presenting cells (APC) pulsed with antigen induces antigen-specific tolerance in both naive and previously primed mice. Although TGF-beta-treated APC-induced tolerance is associated with induction of regulatory T cells and impaired delayed type hypersensitivity (DTH) responses, the specific mechanisms that mediate this tolerance are not currently known. The goal of the present report was to study the mechanisms involved in TGF-beta-treated APC-induced tolerance by determining the fate of the antigen-specific effector T cells that are regulated. Using a well-characterized system that allows tracking of small numbers of TCR transgenic T cells, we have found that antigen-specific T cell expansion, either in vivo or in vitro, is inhibited in mice that have been injected with TGF-beta-treated APC. The failure of antigen-specific effector T cells to expand did not appear to be due to the induction of anergy, since carboxyfluorescein diacetate succinimidyl ester (CFSE)-labeled cells divided normally in response to antigen and adjuvant in vivo, and addition of exogenous IL-2 was unable to restore T cell expansion in in vitro assays. Interestingly, the percentage of CFSE-labeled cells was decreased after >7-8 divisions following culture in vitro, which correlated with a significant increase in cell death. Cell death was prevented and the ability to expand in vitro was restored by treatment with anti-Fas ligand (FasL) antibody. In conclusion, tolerance induced by TGF-beta-treated APC appears to be associated with deletion of antigen-specific T cells involving the Fas-FasL pathway. PMID- 12882833 TI - Membrane lymphotoxin is required for resistance to Theiler's virus infection. AB - Lymphotoxin (LT) and tumor necrosis factor (TNF) are important in immune system development and function. LT consists of soluble LT-alpha(3), which binds to TNF R1 and TNF-R2, and membrane LT-alpha(1)beta(2), which binds to LT-beta-R. We investigated the role of LT and TNF in disease induced by Daniel's (DA) strain of Theiler's murine encephalomyelitis virus (TMEV) since the immune response is believed to be important in both resistance to DA infection as well as mediation of virus-induced demyelination. DA persisted and induced inflammatory demyelination in LT-alpha(-/-) (but not TNF(-/-)) weanling mice of a normally resistant haplotype (C57BL/6), suggesting that LT, but not TNF, is critical for resistance to DA infection. This activity of LT depends on membrane LT alpha(1)beta(2) and not soluble LT-alpha(3), since DA virus persisted and induced inflammatory demyelination in LT-beta-R(-/-), but not TNF-R1(-/-) or TNF-R2(-/-), mice. The LT-alpha(-/-) and LT-beta-R(-/-) mice failed to mount a virus-specific cytotoxic T cell response. Treatment of weanling C57BL/6 mice with LT-beta-R-Ig, which blocks membrane LT activity, failed to increase susceptibility, suggesting that the LT effect is related to its action on immune system development which is fixed by 3 weeks of age. Our data suggest that membrane LT is important in resistance to DA infection (possibly through interference with CD8+ T cell development and function). There was relatively little demyelination associated with inflammation in LT-alpha(-/-) and LT-beta-R(-/-) mice compared to susceptible SJL mice, suggesting the possibility that LT plays a role in mediating demyelination. PMID- 12882834 TI - Histamine deficiency in gene-targeted mice strongly reduces antigen-induced airway hyper-responsiveness, eosinophilia and allergen-specific IgE. AB - Histamine is an important mediator released from activated mast cells provoked by allergen and has a substantial role in the pathophysiology of asthma. However, several lines of evidence indicate that histamine could also have important functions in the regulation of basic cell biological processes. We have used histidine decarboxylase gene-targeted (HDC-KO) mice, lacking histamine, to investigate the effect of histamine deficiency in an animal model of asthma. Our previous investigations revealed that HDC-KO mice had fewer mast cells with reduced granular content and defective degranulation characteristics. Ovalbumin (OVA)-sensitized and challenged HDC-KO mice had significantly reduced airway hyper-responsiveness, lung inflammation, bronchoalveolar lavage eosinophilia, and OVA-specific IgE compared with congenic wild-type littermates treated in the same way. Comparing the expression profiles of cytokines, the levels of IL-1alpha, IL 1beta, IL-4, IL-5, IL-6 and IFN-gamma were significantly lower in the HDC-KO mice in asthmatic late phase, indicating a significantly altered immune response to OVA provocation and challenge. Evaluation of chemokine gene expression revealed that OVA treatment caused elevation of both T(h)1- and T(h)2-type chemokines in wild-type mice, while the chemokine expression was polarized toward a T(h)1 response in HDC-KO mice. According to our results we can suggest that the possible causes of the reduced asthma symptoms in the HDC-KO mice may be the imperfect mast and eosinophil cell system, and an altered immune response to OVA provocation and challenge. PMID- 12882835 TI - Chronic GVH prevents anergy in bone marrow self-reactive B cells: a selective increase in post-endoplasmic reticulum processing and trafficking to the cell surface of autoreactive IgM receptors. AB - B cell autoreactivity is a component of chronic graft versus host (GVH) disease in humans and mice. Chronic GVH driven by I-A disparity results in loss of B cell tolerance in Ig/sHEL tolerant mice. In these mice, B cell anergy is characterized by down-modulation of sIgM mediated by intracellular retention in the endoplasmic reticulum (ER) and/or a block in post-ER processing of IgM receptors. Here, we report that GVH induces a selective increase in post-ER processing of the micro chain and trafficking to the cell surface of IgM receptors in B cells that bind HEL self-antigen. The increase in sIgM was detectable as early as 6 days post GVH, before the appearance of circulating autoantibodies, and was particularly prominent in B cells that up-regulated surface I-A. A further increase in sIgM was found at later time points, along with circulating anti-HEL autoantibodies and a marked decrease in serum-free HEL, but no significant change in the amounts of HEL bound to B cells in vivo. These findings suggest that (i) abrogation of ER retention of IgM receptors in self-reactive B cells is an early event triggered by allogeneic T cells and (ii) at later stages of GVH disease the appearance of autoantibodies reduces the availability of free autoantigen, which may further escalate anergy escape of self-reactive B cells, and lead to exacerbation and perpetuation of autoimmunity. PMID- 12882836 TI - CD69-null mice protected from arthritis induced with anti-type II collagen antibodies. AB - CD69, known as an early activation marker antigen on T and B cells, is also expressed on platelets and activated neutrophils, suggesting certain roles in inflammatory diseases. In order to address the role of CD69 in the pathogenesis of arthritis, we established CD69-null mice. CD69-null mice displayed a markedly attenuated arthritic inflammatory response when injected with anti-type II collagen antibodies. Cell transfer experiments with neutrophils, but not T cells or spleen cells, from wild-type mice into CD69-null mice restored the induction of arthritis. These results indicate a critical role for CD69 in neutrophil function in arthritis induction during the effector phase. Thus, CD69 would be a possible therapeutic target for arthritis in human patients. PMID- 12882837 TI - MHC class II-mediated apoptosis in dendritic cells: a role for membrane associated and mitochondrial signaling pathways. AB - Cytotoxic elimination of dendritic cells (DC) in lymphoid tissue represents an important pathway of immune regulation. However, the mechanism of DC removal is still controversial since mature DC are insensitive to death receptor-mediated killing and other surface or soluble molecules mediating DC death in vivo have yet to be characterized. Class II ligation is the only known signal that induces rapid cell death in mature DC, thus our studies have now focused on the requirements for this cell death using the advantages of tools available for both the mouse and human systems. Anti-class II mAb could be grouped into (i) mAb that both bound to class II and caused class II-mediated cell death as well as (ii) those that bound to class II, but did not cause apoptosis. mAb binding stable class II dimers as well as those mAb recognizing either the alpha or beta chains of class II were found in both groups. Whereas class II-mediated death was enhanced by DC-DC homotypic interactions, DC clustering itself was insufficient to induce apoptosis. Although DC death could be inhibited by uncoupling actin filament bundling, the inhibition of various proteases, including the caspases, and protein transport mediators failed to inhibit class II-mediated cell death. Neither Bid, poly-ADP-ribose polymerase, caspases-3, -7 and -8 nor FLICE inhibitory protein were found to be cleaved during class II apoptosis. Lastly, although class II mAb induced a rapid mitochondrial membrane depolarization in DC, cell death was not inhibited by Bcl-2 over-expression in DC. The independence of this form of apoptosis from protein or RNA synthesis, coupled to the rapidity of the mitochondrial depolarization and the lack of protection by Bcl-2, suggests that mature DC express pre-formed pro-apoptotic molecules that are involved in class II-mediated death. PMID- 12882838 TI - Effect of CXC chemokine platelet factor 4 on differentiation and function of monocyte-derived dendritic cells. AB - Platelet factor 4 (PF4) is a CXC chemokine secreted by activated platelets. PF4 has been shown to promote monocyte survival and induce the differentiation of monocytes into macrophages. However, the effect of PF4 on differentiation of monocytes into dendritic cells (DC) has yet to be determined. As reported previously, monocytes cultured in RPMI medium containing FCS, granulocyte macrophage colony stimulating factor and IL-4 differentiated into CD1a+ DC. When PF4 was added, the expression of CD1a on DC was inhibited. This inhibitory effect was not observed with the other platelet-derived CXC chemokine, beta thromboglobulin. The relative number of CD1a- DC increased from 17 to 92% when the PF4 concentration was increased from 0 to 10 micro g/ml. The inhibitory effect of PF4 on CD1a expression was reversed by 50 U/ml heparin. DC developed in the PF4-containing media appeared more adhesive to plastic culture wells and had higher light side scatter by flow cytometry. Immunophenotypically, monocyte derived DC in the presence of increasing concentrations of PF4 proportionally expressed higher CD86 and lower HLA-DR. The levels of CD11c, CD40 and CD80 remained unchanged with or without PF4. Both CD1a+ DC and CD1a- DC were negative for CD14, CD68 and CD83. Functionally, DC developed in the presence of PF4 had their secretion of tumor necrosis factor-alpha and IL-12 reduced by 75 +/- 10 and 79 +/- 13% respectively when they were stimulated by 100 ng/ml lipopolysaccharide and 50 ng/ml IFN-gamma. CD1a- DC developed in the presence of PF4 were not as active as the control CD1a+ DC in stimulating allogeneic T cells to proliferate. In addition, CD1a- DC were less potent in priming naive CD4+ T cells to secrete both type 1 and 2 cytokines. These results indicate that PF4 can influence differentiation and function of monocyte-derived DC. PMID- 12882839 TI - Retinoic acids exert direct effects on T cells to suppress Th1 development and enhance Th2 development via retinoic acid receptors. AB - The vitamin A metabolite, retinoic acid (RA), affects Th1 and Th2 development. The effect is partly exerted through the modulation of antigen-presenting cell functions, but it remains unclear whether RA directly exerts its effect on T cells to influence Th1/Th2 development. To clarify this problem, we used two experimental systems with isolated T cells in vitro. In one system, isolated CD4+CD8+ thymocytes differentiated into Th1 and Th2 by two transient stimulations with defined combinations of ionomycin and phorbol myristate acetate followed by treatment with IL-2 and IL-4 and/or IL-12. In the second system, functional differentiation was induced in purified naive CD4 T cells from DO-11.10 TCR transgenic and RAG-2-deficient mice with cytokines and antibodies to CD3 and CD28. In both systems, all-trans-RA at > or = 1 nM concentrations suppressed Th1 development, but enhanced Th2 development. 9-cis-RA elicited similar effects. The optimal enhancement of Th2 development in the second system, however, was achieved with a delayed addition of RA. The presence of RA during the initial stimulation period often suppressed Th2 development. The RA receptor (RAR) antagonists, LE540 and LE135, but not the retinoic X receptor (RXR) antagonist, PA452, inhibited the effect of RA on Th1/Th2 development. Accordingly, the RAR agonists, Am80 and Tp80, but not the RXR agonists, HX600 and TZ335, mimicked the effect of RA. The RXR agonists enhanced the effect of the RAR agonists only slightly, if at all. These results indicate that, via RAR, RA directly suppresses Th1 development and directly enhances Th2 development with its timely addition. PMID- 12882840 TI - Involvement of CD100, a lymphocyte semaphorin, in the activation of the human immune system via CD72: implications for the regulation of immune and inflammatory responses. AB - CD100/Sema4D belongs to the semaphorin family, factors known to act as repulsive cues for axons during neuronal development. Mouse CD100 plays a crucial role in both humoral and cellular immunity through ligation of the lymphocyte receptor, CD72. It remains controversial, however, whether human CD100 can function through human CD72 in a manner similar to mouse CD100. To determine the function of human CD100, we generated a recombinant soluble human CD100 protein comprised of the extracellular region of human CD100 fused to the human IgG1 Fc region (hCD100 Fc). hCD100-Fc specifically binds to cells expressing human CD72. As observed previously in the mouse, hCD100-Fc induces the tyrosine dephosphorylation of human CD72, leading to the dissociation of SHP-1 from the CD72 cytoplasmic tail. Consistent with findings for mouse CD100, hCD100-Fc exerts a co-stimulatory effect on B cells and dendritic cells that are stimulated with anti-CD40 mAb. Furthermore, both hCD100-Fc and anti-human CD72 agonistic mAb induce the production of the pro-inflammatory cytokines tumor necrosis factor-alpha, IL-6 and IL-8, even in the absence of anti-CD40 mAb. Collectively, our findings not only demonstrate that human CD100, interacting with human CD72, can function as a ligand in a manner similar to mouse CD100, but also suggest the involvement of human CD100 in inflammatory responses. PMID- 12882841 TI - Beneficial effects of insulin versus sulphonylurea on insulin secretion and metabolic control in recently diagnosed type 2 diabetic patients. AB - OBJECTIVE: To evaluate whether treatment with insulin in recently diagnosed type 2 diabetes is advantageous compared with glibenclamide treatment. RESEARCH DESIGN AND METHODS: Beta-cell function, glycemic control, and quality of life were monitored over 2 years in 39 patients with islet cell antibody-negative type 2 diabetes diagnosed 0-2 years before inclusion in a Swedish multicenter randomized clinical trial. Patients were randomized to either two daily injections of premixed 30% soluble and 70% NPH insulin or glibenclamide (3.5-10.5 mg daily). C peptide-glucagon tests were performed yearly in duplicate after 2-3 days of temporary withdrawal of treatment. RESULTS: After 1 year the glucagon-stimulated C-peptide response was increased in the insulin-treated group by 0.14 +/- 0.08 nmol/l, whereas it was decreased by 0.12 +/- 0.08 nmol/l in the glibenclamide group, P < 0.02 for difference between groups. After 2 years, fasting insulin levels were higher after treatment withdrawal in the insulin-treated versus the glibenclamide-treated group (P = 0.02). HbA(1c) levels decreased significantly during the first year in both groups; however, at the end of the second year, HbA(1c) had deteriorated in the glibenclamide group (P < 0.01), but not in the insulin-treated group. The difference in evolution of HbA(1c) during the second year was significant between groups, P < 0.02. A questionnaire indicated no difference in well-being related to treatment. CONCLUSIONS: Early insulin versus glibenclamide treatment in type 2 diabetes temporarily prolongs endogenous insulin secretion and promotes better metabolic control. PMID- 12882842 TI - Insulin 70/30 mix plus metformin versus triple oral therapy in the treatment of type 2 diabetes after failure of two oral drugs: efficacy, safety, and cost analysis. AB - OBJECTIVE: Subjects (n = 188) with type 2 diabetes and inadequate response to two oral medications (A1C >8.0%) were randomly assigned to treatment with either a third oral medication or an insulin 70/30 mix b.i.d. plus metformin for a comparison of efficacy, safety, and cost. RESEARCH DESIGN AND METHODS: The protocol called for aggressive dose titration to achieve target values of fasting blood glucose (80-120 mg/dl), postprandial glucose (<160 mg/dl), and A1C (<7%). These efficacy parameters were evaluated at weeks 2, 6, 12, and 24 of therapy. If dose adjustments failed to achieve targeted glycemic control, subjects were switched to an alternate therapy. RESULTS: At the end of study (week 24 of therapy), A1C and fasting plasma glucose (FPG) values showed comparable decreases in the two treatment groups. Only 31% (oral therapy) and 32% (insulin plus metformin) of subjects achieved target values of A1C (<7%). A total of 10 of the 98 subjects randomized to triple oral therapy (10.2%) who failed to improve sufficiently were switched to insulin therapy. An additional four subjects dropped out of the oral treatment group due to adverse events felt to be potentially drug related. Only two of the subjects randomized to insulin plus metformin had to be switched to basal-bolus regimens (regular insulin and NPH insulin). Cost analysis determined that insulin plus metformin (mean cost 3.20 dollars/day) provided efficacy equal to that of a triple oral drug regimen (10.40 dollars/day). CONCLUSIONS: Insulin 70/30 mix plus metformin was as effective as triple oral therapy in lowering A1C and FPG values. The triple oral regimen was not as cost effective, and a high percentage of subjects (total of 16.3%) did not complete this regimen due to lack of efficacy or side effects. PMID- 12882843 TI - Adiponectin is present in cord blood but is unrelated to birth weight. AB - OBJECTIVE: In adults, adiponectin is reduced in association with excess adiposity, type 2 diabetes, and hyperinsulinemia. We assessed whether adiponectin was 1) present in the fetal circulation, 2) altered in the fetal circulation in the presence of maternal diabetes, and 3) had relations to fetal cord blood insulin or adiposity. RESEARCH DESIGN AND METHODS: We assessed adiponectin in cord blood in a large cohort of singleton offspring of diabetic mothers (ODM; n = 134) and control mothers (n = 45). RESULTS: Adiponectin was present in cord blood and, in ODM, was higher in those delivered at later gestational ages (Spearman r = 0.18, P = 0.03). Adiponectin was slightly lower in ODM than control subjects (ODM 19.7 +/- 6.1 vs. control 21.8 +/- 5.3 micro g/ml; P = 0.04), although this difference could potentially reflect different gestational ages in the two groups (ODM 37.6 +/- 1.5 and control 40.1 +/- 1.1 weeks). In contrast to adults, adiponectin levels in the fetus were unrelated to the degree of adiposity, blood insulin, or leptin in either control subjects or ODM. CONCLUSIONS: Adiponectin is present in cord blood but does not show expected physiological relations with adiposity as observed in adults. PMID- 12882844 TI - Specialist nurse-led intervention to treat and control hypertension and hyperlipidemia in diabetes (SPLINT): a randomized controlled trial. AB - OBJECTIVE: To determine the effectiveness of specialist nurse-led clinics for hypertension and hyperlipidemia provided for diabetic patients receiving hospital based care. RESEARCH DESIGN AND METHODS: This study was a randomized controlled implementation trial at Hope Hospital, Salford, U.K. The subjects consisted of 1,407 subjects presenting for annual review with raised blood pressure(>or=140/80 mmHg), raised total cholesterol (>or=5.0 mmol/l), or both. Individuals with diabetes were randomized to usual care or usual care with subsequent invitation to attend specialist nurse-led clinics. Nurses provided clinics for participants, with attendance every 4-6 weeks, until targets were achieved. Lifestyle advice and titration of drug therapies were provided according to the locally agreed upon guidelines. Patients with both conditions were eligible for enrollment in either or both clinics. At subsequent annual review, blood pressure and total cholesterol values were obtained from the Salford electronic diabetes register. Data relating to deaths were obtained from the national strategic tracing service. The primary outcome was the odds ratio of achieving targets in hypertension and hyperlipidemia, attributable to the specialist nurse-led intervention. RESULTS: Overall, specialist nurse-led clinics were associated with a significant improvement in patients achieving the target after 1 year (odds ratio [OR] 1.37 [95% CI 1.11-1.69], P = 0.003). This primary analysis revealed a borderline difference in effect between the two types of clinics (test for interaction between groups: P = 0.06). Secondary analysis, consistent with the prior beliefs of the health care professionals involved, suggested that targets were achieved more frequently in patients enrolled in the specialist nurse-led clinic for hyperlipidemia (OR 1.69 [1.25-2.29], P = 0.0007) than for hypertension (OR 1.14 [0.86-1.51], P = 0.37). Intervention (enrolled to either or both clinics) was associated with a reduction in all-cause mortality (OR 0.55 [0.32 0.92], P = 0.02). CONCLUSIONS: This study provides good evidence to support the use of specialist nurse-led clinics as an effective adjunct to hospital-based care of patients with diabetes. If the standards of care recommended in the National Service Framework for Diabetes are to be achieved, then such proven methods for delivering care must be adopted. PMID- 12882845 TI - Management of uncontrolled hypertension in a nurse-led clinic compared with conventional care for patients with type 2 diabetes. AB - OBJECTIVE: To compare the effectiveness of a nurse-led hypertension clinic with conventional community care in general practice in the management of uncontrolled hypertension in patients with type 2 diabetes. RESEARCH DESIGN AND METHODS: We studied 120 men and women outpatient attendees (61% non-Caucasian) with type 2 diabetes and a seated blood pressure (BP) >or=140/80 mmHg. All patients were being treated for hypertension, and 71% had increased urinary albumin excretion (UAE). Patients were allocated to either a nurse-led hypertension clinic or conventional primary care. The primary outcome measure was a change in systolic BP. Secondary outcome measures were total cholesterol, HDL cholesterol, total triglycerides, HbA(1c), UAE, serum creatinine, and changes in absolute stroke and coronary heart disease (CHD) risk scores. RESULTS: The mean (95% CI) difference in the decrement of systolic BP was 12.6 mmHg (5.9-19.3) (P = 0.000) in favor of the nurse-led group, whose patients were three times more likely to a reach target systolic BP <140 mmHg compared with conventional care (P = 0.003). A significant fall in 10-year CHD (P = 0.004) and stroke risk (P = 0.000) scores occurred only in the nurse-led group. There were no significant differences in the reduction of diastolic BP or any of the other secondary outcome measures at 6 months. CONCLUSIONS: Compared with conventional care, a nurse-led hypertension clinic is a more effective intervention for patients with type 2 diabetes and uncontrolled hypertension. A target systolic BP <140 mmHg is more readily achieved and may be associated with significant reductions in 10-year cardiovascular disease risk scores. PMID- 12882846 TI - Low-glycemic index diets in the management of diabetes: a meta-analysis of randomized controlled trials. AB - OBJECTIVE: The use of diets with low glycemic index (GI) in the management of diabetes is controversial, with contrasting recommendations around the world. We performed a meta-analysis of randomized controlled trials to determine whether low-GI diets, compared with conventional or high-GI diets, improved overall glycemic control in individuals with diabetes, as assessed by reduced HbA(1c) or fructosamine levels. RESEARCH DESIGN AND METHODS: Literature searches identified 14 studies, comprising 356 subjects, that met strict inclusion criteria. All were randomized crossover or parallel experimental design of 12 days' to 12 months' duration (mean 10 weeks) with modification of at least two meals per day. Only 10 studies documented differences in postprandial glycemia on the two types of diet. RESULTS: Low-GI diets reduced HbA(1c) by 0.43% points (CI 0.72-0.13) over and above that produced by high-GI diets. Taking both HbA(1c) and fructosamine data together and adjusting for baseline differences, glycated proteins were reduced 7.4% (8.8-6.0) more on the low-GI diet than on the high-GI diet. This result was stable and changed little if the data were unadjusted for baseline levels or excluded studies of short duration. Systematically taking out each study from the meta-analysis did not change the CIs. CONCLUSIONS: Choosing low-GI foods in place of conventional or high-GI foods has a small but clinically useful effect on medium-term glycemic control in patients with diabetes. The incremental benefit is similar to that offered by pharmacological agents that also target postprandial hyperglycemia. PMID- 12882847 TI - Renoprotective effects of adding angiotensin II receptor blocker to maximal recommended doses of ACE inhibitor in diabetic nephropathy: a randomized double blind crossover trial. AB - OBJECTIVE: We evaluated the renoprotective effects as reflected by short-term changes in albuminuria of dual blockade of the renin-angiotensin system (RAS) by adding an angiotensin II receptor blocker (ARB) to treatment with maximal recommended doses of an ACE inhibitor (ACEI) in patients with type 2 diabetes and nephropathy. RESEARCH DESIGN AND METHODS: A total of 20 patients (17 men and 3 women) with type 2 diabetes along with hypertension and nephropathy were enrolled in this double-blind, randomized, two-period, crossover trial of 8 weeks of treatment with the ARB candesartan 16 mg daily and placebo added in random order to existing treatment with lisinopril/enalapril 40 mg daily or captopril 150 mg daily. At the end of each treatment period, we evaluated albuminuria in three 24 h urinary collections by turbidimetry, 24-h ambulatory blood pressure (ABP) using the Takeda-TM2420, and glomerular filtration rate (GFR) by the (51)Cr-EDTA plasma clearance technique. RESULTS: During monoblockade of the RAS by ACEI treatment, albuminuria was 706 (349-1,219) mg/24 h [geometric mean (IQR)]; 24-h ABP was 138 +/- 3/72 +/- 2 mmHg (mean +/- SE); and GFR was 77 +/- 6 ml x min(-1) x 1.73 m(-2) (mean +/- SE). During dual blockade of the RAS by addition of candesartan 16 mg daily, there was a mean (95% CI) reduction in albuminuria of 28 (17-38) compared with ACEI alone (P < 0.001). There was a modest reduction in systolic/diastolic 24-h ABP of 3/2 mmHg (-2 to 8 systolic, -2 to 5 diastolic; NS). Changes in albuminuria did not correlate to changes in ABP. Addition of candesartan 16 mg daily induced a small, insignificant decrease in GFR of 4 (-1 to 9) ml x min(-1) x 1.73 m(-2). CONCLUSIONS: Dual blockade of the RAS provides superior short-term renoprotection independent of systemic blood pressure changes in comparison with maximally recommended doses of ACEI in patients with type 2 diabetes as well as nephropathy. PMID- 12882848 TI - Impact of population management with direct physician feedback on care of patients with type 2 diabetes. AB - OBJECTIVE: Population-level strategies may improve primary care for diabetes. We designed a controlled study to assess the impact of population management versus usual care on metabolic risk factor testing and management in patients with type 2 diabetes. We also identified potential patient-related barriers to effective diabetes management. RESEARCH DESIGN AND METHODS: We used novel clinical software to rank 910 patients in a diabetes registry at a single primary care clinic and thereby identify the 149 patients with the highest HbA(1c) and cholesterol levels. After review of the medical records of these 149 patients, evidence-based guideline recommendations regarding metabolic testing and management were sent via e-mail to each intervention patient's primary care provider (PCP). Over a 3 month follow-up period, we assessed changes in the evidence-based management of intervention patients compared with a matched cohort of control patients receiving usual care at a second primary care clinic affiliated with the same academic medical center. RESULTS: In the intervention cohort, PCPs followed testing recommendations more often (78%) than therapeutic change recommendations (36%, P = 0.001). Compared with the usual care control cohort, population management resulted in a greater overall proportion of evidence-based guideline practices being followed (59 vs. 45%, P = 0.02). Most intervention patients (62%) had potential barriers to effective care, including depression (35%), substance abuse (26%), and prior nonadherence to care plans (18%). CONCLUSIONS: Population management with clinical recommendations sent to PCPs had a modest but statistically significant impact on the evidence-based management of diabetes compared with usual care. Depression and substance abuse are prevalent patient level adherence barriers in patients with poor metabolic control. PMID- 12882849 TI - Effect of nurse-directed diabetes care in a minority population. AB - OBJECTIVE: To determine whether diabetes care directed by nurses following detailed protocols and algorithms and supervised by a diabetologist results in meeting the evidence-based American Diabetes Association (ADA) process and outcome measures more often than care directed under usual care in a minority population. RESEARCH DESIGN AND METHODS: Studies were mainly conducted in two Los Angeles County clinics. In clinic A, nurse-directed diabetes care was provided to 252 patients (92% Hispanic and 2% African-American) referred by their primary care providers. These patients were hierarchically matched with 252 diabetic patients in clinic B (79% Hispanic and 19% African American). When nurse-directed care was abruptly discontinued in clinic A for administrative reasons, it was reestablished in clinic B. Those patients were randomly selected from a teaching clinic, and the outcomes in 114 patients who completed 1 year were compared with outcomes derived the year before receiving nurses' care. The following process and outcome measures were assessed in the study: 1) number of visits, 2) diabetes education, 3) nutritional counseling, 4) HbA(1c), 5) lipid profiles, 6) eye exams, 7) foot exams, 8) renal evaluations, and 9) ACE inhibitor therapy in appropriate patients. RESULTS: For patients under nurse-directed diabetes care in both clinics A and B, almost all process measures were carried out significantly more frequently than for the appropriate control patients. Under the care of nurses in clinic A, HbA(1c) levels fell 3.5% from 13.3 to 9.8% in the 120 patients who were followed for at least 6 months, as compared with a 1.5% fall from 12.3 to 10.8% under usual (physician-directed) care in clinic B. During the year before enrolling in nurse-directed care in clinic B, mean HbA(1c) levels decreased from 10.0 to 8.5%. At the end of a year under the nurses' care, the values fell further to 7.1%. The median value fell from 8.3 to 6.6%. CONCLUSIONS: Specially trained nurses who follow detailed protocols and algorithms under the supervision of a diabetologist can markedly improve diabetes outcomes in a minority population. This approach could help blunt the increased morbidity and mortality noted in minority populations. PMID- 12882850 TI - Biologic and quality-of-life outcomes from the Mediterranean Lifestyle Program: a randomized clinical trial. AB - OBJECTIVE: Few multiple lifestyle behavior change programs have been designed to reduce the risk of coronary heart disease in postmenopausal women with type 2 diabetes. This study tested the effectiveness of the Mediterranean Lifestyle Program (MLP), a comprehensive lifestyle self-management program (Mediterranean low-saturated fat diet, stress management training, exercise, group support, and smoking cessation), in reducing cardiovascular risk factors in postmenopausal women with type 2 diabetes. RESEARCH DESIGN AND METHODS: Postmenopausal women with type 2 diabetes (n = 279) were randomized to either usual care (control) or treatment (MLP) conditions. MLP participants took part in an initial 3-day retreat, followed by 6 months of weekly meetings, to learn and practice program components. Biological end points were changes in HbA(1c), lipid profiles, BMI, blood pressure, plasma fatty acids, and flexibility. Impact on quality of life was assessed. RESULTS: Multivariate ANCOVAs revealed significantly greater improvements in the MLP condition compared with the usual care group on HbA(1c), BMI, plasma fatty acids, and quality of life at the 6-month follow-up. Patterns favoring intervention were seen in lipids, blood pressure, and flexibility but did not reach statistical significance. CONCLUSIONS: These results demonstrate that postmenopausal women with type 2 diabetes can make comprehensive lifestyle changes that may lead to clinically significant improvements in glycemic control, some coronary heart disease risk factors, and quality of life. PMID- 12882851 TI - Out-of-pocket costs and diabetes preventive services: the Translating Research Into Action for Diabetes (TRIAD) study. AB - OBJECTIVE: Despite the increased shifting of health care costs to consumers, little is known about the impact of financial barriers on health care utilization. This study investigated the effect of out-of-pocket expenditures on the utilization of recommended diabetes preventive services. RESEARCH DESIGN AND METHODS: This was a survey-based observational study (2000-2001) in 10 managed care health plans and 68 provider groups across the U.S. serving approximately 180,000 patients with diabetes. From 11,922 diabetic survey respondents, we studied the occurrence of self-reported annual dilated eye exams and diabetes health education and among insulin users, daily self-monitoring of blood glucose (SMBG). Conditional probabilities were estimated for outcomes at each level of self-reported out-of-pocket expenditure by using hierarchical logistic regression models with random intercepts. RESULTS: Conditional probabilities of utilization (95% CI) varied by expenditure for dilated eye exam [no cost 78% (75-82), copay 79% (75-82), and full price 70% (64-75); P < 0.0001]; diabetes health education [no cost 29% (23-36), copay 29% (23-36), and full price 19% (14-25); P < 0.0001]; and daily SMBG [no cost 75% (68-81), copay 68% (60-75), and full price 59% (49 68); P < 0.0001]. Extensive adjustment for patient factors had no discernible effect on the estimates or their significance, and cost-utilization relationships were similar across income levels and other patient characteristics. CONCLUSIONS: Benefit packages structured to derive greater fiscal contribution from the health plan membership result in suboptimal use of diabetes preventive services and may thus lead to poorer clinical outcomes, greater future costs, and lower health plan quality ratings. PMID- 12882852 TI - The direct medical cost of type 2 diabetes. AB - OBJECTIVE: To describe the direct medical costs associated with type 2 diabetes, as well as its treatments, complications, and comorbidities. RESEARCH DESIGN AND METHODS: We studied a random sample of 1,364 subjects with type 2 diabetes who were members of a Michigan health maintenance organization. Demographic characteristics, duration of diabetes, diabetes treatments, glycemic control, complications, and comorbidities were assessed by surveys and medical chart reviews. Annual resource utilization and costs were assessed using health insurance claims. The log-transformed annual direct medical costs were fitted by multiple linear regression to indicator variables for demographics, treatments, glycemic control, complications, and comorbidities. RESULTS: The median annual direct medical costs for subjects with diet-controlled type 2 diabetes, BMI 30 kg/m(2), and no microvascular, neuropathic, or cardiovascular complications were 1,700 dollars for white men and 2,100 dollars for white women. A 10-kg/m(2) increase in BMI, treatment with oral antidiabetic or antihypertensive agents, diabetic kidney disease, cerebrovascular disease, and peripheral vascular disease were each associated with 10-30% increases in cost. Insulin treatment, angina, and MI were each associated with 60-90% increases in cost. Dialysis was associated with an 11-fold increase in cost. CONCLUSIONS: Insulin treatment and diabetes complications have a substantial impact on the direct medical costs of type 2 diabetes. The estimates presented in this model may be used to analyze the cost-effectiveness of interventions for type 2 diabetes. PMID- 12882853 TI - Predicted costs and outcomes from reduced vibration detection in people with diabetes in the U.S. AB - OBJECTIVE: The ability to perceive vibration (vibration detection) has been shown to be a good predictor of the long-term complications of diabetic peripheral neuropathy (DPN). We aimed to estimate the predicted complications and costs for the U.S. health care system associated with reduced vibration detection (vibration perception threshold >or=25 V), estimated using a quantitative sensory testing device. RESEARCH DESIGN AND METHODS: A Markov model was constructed for a hypothetical cohort of people with DPN. The model was run over a 10-year period using Monte Carlo simulations to estimate disease progression, predicted costs, and complications according to vibration detection levels. RESULTS: The average individual with reduced vibration detection incurs approximately five times more direct medical costs for foot ulcer and amputations, yields 0.18 fewer quality adjusted life-years, and lives for approximately 2 months less than an average individual with normal vibration detection. CONCLUSIONS: The treatment of foot ulceration and amputation is time-consuming and expensive. If individuals with reduced vibration detection could be identified, then preventative care could be concentrated on those patients, potentially saving valuable resources and improving health outcomes. PMID- 12882854 TI - A descriptive and comparative study of the prevalence of depressive and anxiety disorders in low-income adults with type 2 diabetes and other chronic illnesses. AB - OBJECTIVE: To determine whether type 2 diabetes contributes to the presence of depressive and anxiety disorder diagnoses in low-income adults with hypertension, asthma, and/or arthritis. RESEARCH DESIGN AND METHODS: Using a cross-sectional design, this study administered a structured diagnostic interview to low-income primary care patients diagnosed with type 2 diabetes, hypertension, arthritis, and asthma, as well as to those with no chronic illness (n = 326), to determine the 12-month prevalence of depressive and anxiety disorders. A logistic regression (LR) model was used to assess whether a diagnosis of depression and/or anxiety was associated with type 2 diabetes after adjusting for known risk factors. RESULTS: A high prevalence rate of depressive and/or anxiety disorders was found in the total sample (29%) and in all three illness groups: type 2 diabetes (36%), other chronic illnesses (24%), and no chronic illness (31%). Using LR, a main effect was detected for illness group when age and education were controlled (chi(2) = 22.66, df 4, P = 0.000). Specifically, the odds of occurrence of a depressive and/or anxiety disorder in those with comorbid type 2 diabetes were twice that in the nondiabetic, chronically ill comparison group (odds ratio 2.26, 95% CI 1.28-4.01, P = 0.005). CONCLUSIONS: These results suggest a positive contribution of type 2 diabetes to increased rates of depressive and/or anxiety disorders in patients with hypertension, asthma, and/or arthritis and support prior research that type 2 diabetes may serve as an indicator of depression and anxiety in low-income adults treated in primary care clinics. PMID- 12882855 TI - Impaired fasting glucose and impaired glucose tolerance in women with prior gestational diabetes are associated with a different cardiovascular profile. AB - OBJECTIVE: The purpose of this study was to investigate the association of cardiovascular risk factors to impaired glucose tolerance (IGT) and to impaired fasting glucose (IFG) in women with prior gestational diabetes mellitus (GDM). RESEARCH DESIGN AND METHODS: We studied 838 women with prior GDM. Postpartum glucose tolerance status was classified as normal, IFG, IGT, IFG plus IGT, and diabetes according to the World Health Organization criteria. Postpartum BMI, waist circumference, blood pressure, triglyceride, cholesterol, and HDL cholesterol were assessed. RESULTS: BMI and blood pressure were significantly higher in women with IFG than in women with normal glucose status. BMI and waist circumference were significantly higher in women with IFG plus IGT than in women with normal glucose status. No differences were observed between women with IGT and normal glucose status. The prevalence of hypertension and obesity was significantly increased in IFG compared with normal glucose status. The prevalence of obesity and abnormal lipids was significantly increased in IFG plus IGT compared with normal glucose status. IGT showed no increased prevalence of cardiovascular risk factors. CONCLUSIONS: Traditional cardiovascular risk factors have a stronger association with isolated IFG than with isolated IGT in women with prior GDM. PMID- 12882856 TI - C-reactive protein predicts the deterioration of glycemia in chinese subjects with impaired glucose tolerance. AB - OBJECTIVE: Recent studies have shown that C-reactive protein (CRP) predicts future risk of diabetes in healthy Caucasians. We determined whether plasma CRP level was elevated in Chinese subjects with impaired glucose tolerance (IGT) and whether CRP level could be used to predict progression to type 2 diabetes or reversion to normal glucose tolerance (NGT) in these high-risk individuals. RESEARCH DESIGN AND METHODS: A total of 228 subjects with IGT at baseline from the Hong Kong Cardiovascular Risk Factors Prevalence Study underwent repeat oral glucose tolerance testing after 2 years. Plasma high-sensitivity CRP was measured from their stored baseline samples and from 228 subjects with NGT matched for age and BMI by an immunoturbidimetric assay. RESULTS: Subjects with IGT at baseline had higher plasma CRP levels than subjects with NGT: 1.18 mg/l (0.52-2.52) vs. 0.87 mg/l (0.37-1.84), median (interquartile range), P = 0.01. At 2 years, 117 subjects with IGT reverted to NGT, 84 remained in IGT, and 21 progressed to diabetes. Individuals who progressed to diabetes had the highest plasma CRP levels at baseline (P < 0.0001). Those with baseline CRP levels in the third and top quartile had a relative risk of remaining in IGT or progressing to diabetes of 2.87 (95% CI 1.06-7.82) and 2.76 (1.06-7.31), respectively, after adjusting for anthropometric measure and lifestyle factors. CONCLUSIONS: CRP independently predicts the risk of remaining in IGT or progressing to diabetes in Chinese subjects with IGT. CRP might provide an adjunctive measure for identifying subjects with the highest risk of progression to diabetes who would derive the greatest benefits from preventive interventions. PMID- 12882857 TI - Diabetes and driving mishaps: frequency and correlations from a multinational survey. AB - OBJECTIVE: The intensive treatment of diabetes to achieve strict glycemic control is a common clinical goal, but it is associated with an increased incidence of hypoglycemia. Becoming hypoglycemic while driving is a hazardous condition and may lead to a greater incidence of driving mishaps. This study investigated whether diabetes is associated with increased risk of driving mishaps and correlates of such a relationship. RESEARCH DESIGN AND METHODS: During routine visits to diabetes specialty clinics in seven U.S. and four European cities, consecutive adults with type 1 diabetes, type 2 diabetes, and nondiabetic spouse control subjects (n = 341, 332, and 363, respectively) completed an anonymous questionnaire concerning diabetes and driving. RESULTS: Type 1 diabetic drivers reported significantly more crashes, moving violations, episodes of hypoglycemic stupor, required assistance, and mild hypoglycemia while driving as compared with type 2 diabetic drivers or spouse control subjects (P < 0.01-0.001). Type 2 diabetic drivers had driving mishap rates similar to nondiabetic spouses, and the use of insulin or oral agents for treatment had no effect on the occurrence of driving mishaps. Crashes among type 1 diabetic drivers were associated with more frequent episodes of hypoglycemic stupor while driving, less frequent blood glucose monitoring before driving, and the use of insulin injection therapy as compared with pump therapy. One-half of the type 1 diabetic drivers and three quarters of the type 2 diabetic drivers had never discussed hypoglycemia and driving with their physicians. CONCLUSIONS: Type 1 diabetic drivers are at increased risk for driving mishaps, but type 2 diabetic drivers, even on insulin, appear not to be at a higher risk than nondiabetic individuals. Clinical and treatment factors appear to increase risk, e.g., more frequent hypoglycemia while driving, method of insulin delivery, and infrequent self-testing before driving. Physicians are encouraged to talk to their type 1 diabetic patients about hypoglycemia and driving. PMID- 12882858 TI - Prevalences of diabetes and impaired glucose regulation in a Danish population: the Inter99 study. AB - OBJECTIVE: To determine the age- and sex-specific prevalence of impaired fasting glycemia, impaired glucose tolerance, screen-detected diabetes, and known diabetes in a Danish population aged 30-60 years and to examine the phenotype and the cardiovascular risk profile in individuals with impaired glucose regulation. RESEARCH DESIGN AND METHODS: In the Inter99 study, 13,016 inhabitants living in Copenhagen County were invited. All participants underwent anthropometric measurements, blood samples, and a 75-g standardized oral glucose tolerance test. RESULTS: The age-specific prevalences in men were as follows: impaired fasting glycemia: 1.4-16.3%; impaired glucose tolerance: 6.9-17.8%; screen-detected diabetes: 0.7-9.7%; and known diabetes: 0-5.8%. The corresponding figures in women were 0-5.1, 10.5-17.3, 0.6-6.3, and 0.5-9%. The prevalence of impaired glucose regulation increased with age. Among individuals with diabetes, 65.6% were previously undiagnosed; this proportion was highest in the youngest age group (82% among 45-year-old men vs. 63% among 60-year-old men, and 70% among 45 year-old women vs. 52% among 60-year-old women). Mean BMI, waist, HbA(1c), systolic blood pressure, diastolic blood pressure, and total cholesterol were significantly higher (P < 0.0001) in the individuals with impaired glucose regulation compared with individuals with normal glucose tolerance. CONCLUSIONS: This study revealed that the prevalence of type 2 diabetes is high and that still two out of three individuals are undiagnosed, indicating a need for more attention to the disease in society. PMID- 12882859 TI - Increased visceral fat and serum levels of triglyceride are associated with insulin resistance in Japanese metabolically obese, normal weight subjects with normal glucose tolerance. AB - OBJECTIVE: The purpose of this study was to investigate the association between visceral adiposity or triglyceride (TG) metabolism and insulin resistance in metabolically obese, normal weight (MONW) Japanese individuals with normal glucose tolerance. RESEARCH DESIGN AND METHODS: We evaluated body fat areas, lipid profiles, and the glucose infusion rate (GIR) during a euglycemic hyperinsulinemic clamp study in 20 MONW subjects (BMI <25 kg/m(2)and visceral fat areas 100 cm(2)) with normal glucose tolerance. Body fat areas were measured by computed tomography scans. Control data were obtained from 20 normal subjects (BMI <25 kg/m(2) and visceral fat areas <100 cm(2)). RESULTS: MONW subjects showed a significant increase in fasting serum levels of TG (P < 0.01) and a decrease in GIR (P < 0.01) compared with normal subjects. There were significant correlations between visceral fat areas (r = -0.563, P < 0.01) or serum levels of TG (r = -0.474, P < 0.05) and GIR in MONW subjects. Multiple regression analyses showed that visceral fat areas (F = 7.702, P < 0.02) and serum levels of TG (F = 7.114, P < 0.05) were significantly associated with GIR in all (MONW and normal) subjects. CONCLUSIONS: Increased visceral fat and serum levels of TG are associated with insulin resistance in Japanese MONW subjects with normal glucose tolerance. Excess visceral fat and elevated TG levels may play important roles in the development of insulin resistance in Japanese MONW subjects with normal glucose tolerance. PMID- 12882860 TI - Fulminant type 1 diabetes: a nationwide survey in Japan. AB - OBJECTIVE: To describe the clinical and immunologic characteristics of fulminant type 1 diabetes, a novel subtype of type 1 diabetes, we conducted a nationwide survey. RESEARCH DESIGN AND METHODS: History and laboratory data, including islet related autoantibodies, were examined in 222 patients with fulminant and nonfulminant type 1 diabetes in our hospitals in addition to another 118 patients with fulminant type 1 diabetes located outside our hospitals in Japan. RESULTS: In our hospitals, of the 222 patients studied, 43 (19.4%) were diagnosed with fulminant type 1 diabetes, 137 (61.7%) were classified as having autoimmune type 1 diabetes, and 42 were type 1 diabetic subjects who were not fulminant and did not have anti-islet antibodies. An additional 118 fulminant patients outside our hospitals were enrolled, making a total of 161 fulminant type 1 diabetic subjects (83 male and 78 female subjects; 14 children/adolescents and 147 adults) identified from all over Japan. (In 2000, the average incidence was three cases per month.) Flu-like symptoms and pregnancy were more frequently observed in the fulminant than in the autoimmune group (P < 0.001). In the fulminant patients, 4.8% were positive for anti-GAD antibodies and none were positive for anti-islet antigen 2 antibodies. CONCLUSIONS: Fulminant type 1 diabetes is a distinct subtype and accounts for approximately 20% of the ketosis-onset type 1 diabetes cases in Japan. Flu-like symptoms are characteristic of disease onset. Metabolic derangement is more severe in this subtype than in autoimmune type 1 diabetes. PMID- 12882861 TI - Low incidence of end-stage renal disease and chronic renal failure in type 2 diabetes: a 10-year prospective study. AB - OBJECTIVE: Data on the incidence of end-stage renal disease (ESRD) and chronic renal failure from population-based studies in Caucasian type 2 diabetic patients are lacking. To provide such data, a population-based cohort of type 2 diabetic patients was identified in Casale Monferrato, Italy, and prospectively examined from 1991 to 2001. RESEARCH DESIGN AND METHODS: During the follow-up period, patients were regularly examined with centralized measurements of plasma creatinine and HbA(1c). Independent predictors of progression to renal events were identified with multivariate Cox proportional hazards modeling, with sex, age, and individual follow-up time as confounders. RESULTS: We followed 1,408 of 1,540 (91.4%) patients (average follow-up time 6.7 years, range 0.011-11.1); 10 new cases of ESRD and 72 of chronic renal failure (plasma values of creatinine >or=2.0 mg/dl) were identified, giving incidence rates/1,000 person-years of 1.04 (95% CI 0.56-1.94) and 7.63 (6.06-9.61), respectively. Cumulative risks for chronic renal failure adjusted for competing mortality were 6.1 and 9.3% after 20 and 30 years from diagnosis of diabetes, respectively. Incidence rates and cumulative risks of chronic renal failure defined by plasma creatinine values >1.5 mg/dl increased to 13.1/1,000 person-years, 8.6 and 14.8%, respectively. In Cox regression analysis, predictors of progression (after adjustment for confounders) were hypertension (P = 0.078), diastolic blood pressure (P = 0.034), BMI (P = 0.03), and albumin excretion rate (AER) (P < 0.0001). CONCLUSIONS: We provide evidence that the individual risk of ESRD and chronic renal failure is low. AER and diastolic blood pressure are independent predictors of progression. These findings underline the relevance of primary prevention to reduce the number of diabetic patients with ESRD. PMID- 12882862 TI - A comparison of postprandial and preprandial administration of insulin aspart in children and adolescents with type 1 diabetes. AB - OBJECTIVE: The aim of this study was to compare the glycemic control of preprandial versus postprandial injections of the new rapid-acting insulin analogue aspart in children and adolescents with type 1 diabetes. RESEARCH DESIGN AND METHODS: Forty-two children (aged 6-12 years) and 34 adolescents (13-17 years) were randomized to preprandial (immediately before meal start) and postprandial (immediately after a meal or a maximum of 30 min after meal start) treatment with insulin aspart (at least thrice daily) as part of a basal/bolus regimen in a multicenter study with an open labeled, two-period cross-over design (6-week periods). Of this group, 49% were boys, 55% were aged or=60 years, respectively), as well as by glucose tolerance status. RESULTS: Adiponectin concentrations did not differ by age-groups. There were significant age effects for BMI, percent body fat, visceral fat, subcutaneous abdominal fat, VO(2max), and M. Adiponectin levels were lower in the prediabetic women (n = 18) than in the normal glucose-tolerant women (n = 108) and the women with type 2 diabetes (n = 22) (both P < 0.05). Univariate correlations revealed significant negative relationships between plasma adiponectin levels and BMI, percent body fat, visceral fat, subcutaneous abdominal fat, fasting leptin, and fasting insulin and positive relationship with M (all P < 0.05). In a multiple stepwise regression model to predict adiponectin, only M remained in the model at P < 0.001. Multivariate analyses revealed a significant relation for M as a function of adiponectin, insulin, and VO(2max). CONCLUSIONS: The data suggest that plasma adiponectin does not change with age but levels are negatively associated with percent body fat, visceral fat, subcutaneous abdominal fat, insulin, and leptin levels in women. Adiponectin is positively associated with M across the age span in women. PMID- 12882867 TI - Microalbuminuria and mortality in long-duration type 1 diabetes. AB - OBJECTIVE: Microalbuminuria is a recognized risk factor for increased mortality and renal failure in type 1 diabetes. Whether it remains a powerful predictor in patients with a long duration of type 1 diabetes is not known. We ascertained the prognostic significance of abnormal urinary albumin excretion in a cohort of patients with at least 30 years of type 1 diabetes. RESEARCH DESIGN AND METHODS: A total of 190 patients with a disease duration of type 1 diabetes of at least 30 years with baseline and 5 years of follow-up. RESULTS: At baseline 66% were normoalbuminuric, and at 5 years 11% of this cohort had died. Of the 22% who were microalbuminuric at baseline, 26% had died, and of the 8% with persistent proteinuria at baseline, 44% had died. Of the 4% with end-stage renal failure at baseline, 71% had died within 5 years. Death was attributable to a cardiovascular cause in two-thirds of the cases in all groups. CONCLUSIONS: Even in those with a long duration of type 1 diabetes, the presence of abnormal urinary excretion remains a powerful predictor of increased mortality. PMID- 12882868 TI - Racial differences in diabetic nephropathy, cardiovascular disease, and mortality in a national population of veterans. AB - OBJECTIVE: To determine racial differences in the prevalence of diabetic nephropathy, cardiovascular disease (CVD), and risk of mortality in a national health care system. RESEARCH DESIGN AND METHODS: A longitudinal cohort study was conducted in 429,918 veterans with diabetes. Racial minority groups were analyzed for baseline differences in prevalence of early diabetic nephropathy, diabetic end-stage renal disease (ESRD) and CVD, and longitudinal risk of mortality compared with Caucasians. RESULTS: The 429,918 patients identified with diabetes were of the following racial groups: Caucasian (56.2%), African American (15.3%), Asian (0.5%), Native American (0.4%), and unknown race (21.4%). Minority individuals were, on average, younger and less likely to have CVD but were more likely to have renal disease than Caucasians. After adjustment for age, sex, and economic status, African Americans (adjusted odds ratio [OR] = 1.3, 95% CI 1.2 1.4) and Native Americans (1.5, 1.1-2.1) were more likely to have early diabetic nephropathy than Caucasians. Diabetic ESRD was more likely to be present in African Americans (1.9, 1.9-2.0), Hispanics (1.4, 1.3-1.4), Asians (1.8, 1.5 2.1), and Native Americans (1.9, 1.5-2.3) than Caucasians. Concurrently, the adjusted OR of CVD in racial minority groups was 27-49% less than in Caucasians, whereas the 18-month risk of mortality among people from most racial minority groups was 7-12% lower than in Caucasians. CONCLUSIONS: We conclude that when access to care is comparable, microvascular complications, macrovascular disease, and subsequent death occur with different frequencies among various racial groups. PMID- 12882869 TI - Optimal blood glucose control during 18 years preserves peripheral nerve function in patients with 30 years' duration of type 1 diabetes. AB - OBJECTIVE: To assess the association between 18 years of mean HbA(1c) and nerve conduction parameters of the lower limb in patients with type 1 diabetes of 30 years' duration. RESEARCH DESIGN AND METHODS: HbA(1c) has been examined prospectively since 1982 in a group of 39 patients with type 1 diabetes. Mean age at baseline was 25 years (range 18-40) with 12 years' disease duration. The mean age at diagnosis of diabetes was 12.5 years. Nerve function of lower limbs was assessed at baseline, after 8 years, and after 18 years. RESULTS: A total of 23 men and 16 women were studied. Mean age was 43 years. Mean HbA(1c) was 8.2% (range 6.6-11.3) during 18-year follow-up. Nerve conduction velocity (NCV) and nerve action potential amplitude (NAPA) at the last examination were significantly associated with mean HbA(1c) (P < 0. 05). From 1982 to 1999, there was a significant reduction in nerve function in patients with mean HbA(1c) >or=8.4% (highest tertile). For example, the mean NCV in the tibial nerve was reduced from 47 to 31 m/s (P < 0.01). The number of nerves with NCV (P < 0.01) and NAPA (P = 0.01) reduced to below the reference level in each patient was also significantly associated to mean HbA(1c). No significant associations were found between nerve function parameters, sex, disease duration, blood pressure, serum cholesterol, microalbuminuria, or smoking. CONCLUSIONS: The present study shows that mean HbA(1c) is a strong predictor of nerve function. Mean HbA(1c) <8.4% over 18 years was associated with near-normal nerve function. PMID- 12882870 TI - Physiological differences between interstitial glucose and blood glucose measured in human subjects. AB - OBJECTIVE: This study investigated whether glucose readings from a sensor sampling in interstitial fluid differ substantially from blood glucose (BG) values measured at the same time. RESEARCH DESIGN AND METHODS: We have evaluated the relationship between BG and glucose extracted from interstitial fluid using the GlucoWatch (Cygnus, Redwood City, CA) biographer, a device that collects glucose from subcutaneous interstitial space through intact skin by application of a low electric current. We evaluated the relative change in the interstitial glucose (IG) signal (IGS) as measured by the biographer versus BG using a normalized two-point sensitivity index (NSI). RESULTS: The results show that biographer measures of IG differ in time and magnitude from the corresponding BG values. In particular, the biographer values were shifted in time due to instrumental and physiological lag. Results show an average total lag of 17.2 +/- 7.2 min for all subjects evaluated. The instrumental lag was 13.5 min, suggesting that physiological lag is approximately 5 min. In addition, when glucose was increasing, the change in IGS was less than that in BG, while when BG was decreasing, the change in IGS was greater than that in BG. CONCLUSIONS: Similar results have been reported for other measures of IG, suggesting that differences reflect physiological variation in glucose uptake, utilization, and elimination in blood and interstitial space. This further evidence of the difference between IG and BG should be considered when interpreting glucose measurements from devices that sample interstitial fluid. PMID- 12882871 TI - Phenotypic heterogeneity and associations of two aldose reductase gene polymorphisms with nephropathy and retinopathy in type 2 diabetes. AB - OBJECTIVE: We investigated the phenotypic features of diabetic microvascular complications and their association with a (CA)(n) microsatellite and a C/T polymorphism at the 5' region of the aldose reductase gene (ALR2) in a consecutive cohort of 738 Chinese type 2 diabetic patients. RESEARCH DESIGN AND METHODS: Of the entire patient cohort, 392 were free of diabetes complications, or uncomplicated, 159 had diabetic nephropathy, 66 had diabetic retinopathy, and 121 had both diabetic nephropathy and retinopathy. Nephropathy was defined as urinary albumin excretion rate (AER) >or=20 micro g/min and albumin-to-creatinine ratio >or=3.5 mg/mmol in two urine collections. Retinopathy was defined by the presence of hemorrhages, exudates, laser marks, and fibrous proliferation or by a history of vitrectomy. (CA)(n) and C/T polymorphisms were examined by PCR followed by capillary electrophoresis and digestion with BfaI, respectively. RESULTS: In the whole cohort, patients with diabetic retinopathy (n = 187) had higher blood pressure and lower BMI, while those with diabetic nephropathy (n = 280) had higher blood pressure, waist-to-hip ratio, and lipid profile than those without the respective complications. The z+6 carriers of the (CA)(n) polymorphism were less common in patients with diabetic retinopathy than those without diabetic retinopathy (n = 551) (4.3 vs. 9.3%, P = 0.04). The CT/TT carriers had a higher AER than the CC carriers (30.2 x/divided by 7.2 vs. 21.9 x/divided by 6.9 micro g/min, P = 0.03). Further subgroup analysis was performed after excluding uncomplicated patients with <5 years disease duration. The group with both diabetic nephropathy and retinopathy had higher frequencies of the z-2 allele (25.7 vs. 16.9%, P = 0.03) and T allele (26.4 vs. 18.5%, P = 0.04) and a lower frequency of the z+6 allele (1.7 vs. 5.5%, P = 0.054) than the uncomplicated group. Multiple logistic regression analysis confirmed that z-2 carrying (odds ratio 2.6, 95% CI 1.20-5.83, P = 0.02) and CT/TT genotypes (OR 2.5, 95% CI 1.19-5.19, P = 0.02) were independent predictors for both diabetic nephropathy and retinopathy. CONCLUSIONS: Chinese type 2 diabetic patients exhibited phenotypic differences in terms of risk factors for both diabetic nephropathy and diabetic retinopathy. Both the z-2 allele of (CA)(n) polymorphism and T allele of ALR2 were independently associated with severe diabetic microvascular complications. PMID- 12882872 TI - APOE polymorphism and the progression of diabetic nephropathy in Japanese subjects with type 2 diabetes: results of a prospective observational follow-up study. AB - OBJECTIVE: The aim of this study is to clarify the conflicting results of the epsilon2/epsilon3/epsilon4 APOE polymorphism as a risk factor on diabetic nephropathy by a cohort study. RESEARCH DESIGN AND METHODS: A total of 429 Japanese subjects with type 2 diabetes and with normoalbuminuria (n = 299) or with microalbuminuria (n = 130) were enrolled in a prospective observational follow-up study during 1995-1998 and followed until 2001 (for at least 3 years). The endpoint was the occurrence of a renal event defined as the progression to a higher stage of diabetic nephropathy. RESULTS: During the study (the mean follow up period: 4.4 +/- 1.0 years), 31 of 429 subjects progressed: 21 from normoalbuminuria to microalbuminuria and 10 from microalbuminuria to overt proteinuria. The allele frequency of the APOE polymorphism was significantly different between the progressors and the nonprogressors. Eight of 42 epsilon2 carriers (19%) progressed, whereas 23 of 387 noncarriers (6%) progressed with a relative risk of 3.2 (95% CI 1.5-6.7). When subjects were stratified by renal status at baseline, each relative risk for the progression in the epsilon2 carriers was 2.7 (0.99-7.4) in those with normoalbuminuria and 4.2 (1.3-13.3) in those with microalbuminuria. Furthermore, when analyzed only in subjects with normoalbuminuria and short duration of diabetes (<15 years) at baseline, the risk in the epsilon2 carriers became higher to 3.2 (1.2-8.8). CONCLUSIONS: Our follow up study indicates that the epsilon2 allele of the APOE polymorphism is a prognostic risk factor for both the onset and the progression of diabetic nephropathy in Japanese type 2 diabetes. PMID- 12882873 TI - ACE inhibitors improve diabetic nephropathy through suppression of renal MCP-1. AB - OBJECTIVE: Chemokines play an important role in the pathogenesis of diabetic nephropathy. Angiotensin II induces several fibrogenic chemokines, namely monocyte chemoattractant protein-1 (MCP-1) and transforming growth factor-beta. The progression of diabetic nephropathy can be retarded by ACE inhibitors (ACEIs) in patients with type 1 and type 2 diabetes. We examined if blockade of the renin angiotensin system lowered urinary levels of the chemokine MCP-1 and correlated urinary MCP-1 (uMCP-1) with parameters of renal function and glucose and lipid metabolism before and after 1 year of treatment with an ACE inhibitor. RESEARCH DESIGN AND METHODS: In 22 patients with type 2 diabetes and diabetic nephropathy in stages 3-5, treatment with the ACEI lisinopril was initiated. Before treatment and after 12 months of continuous therapy, proteinuria, creatinine clearance, uMCP-1 levels, BMI, HbA(1c), and serum cholesterol were assessed. RESULTS: Lisinopril treatment improved renal function. Proteinuria decreased from 410 +/- 662 mg per 24 h to 270 +/- 389 mg per 24 h. Creatinine clearance rose from 61 +/- 26 to 77 +/- 41 ml/min. Urinary MCP-1 levels decreased from 0.456 +/- 0.22 ng/mg creatinine to 0.08 +/- 0.096 ng/mg creatinine. The change in uMCP-1 correlated significantly (r = 0.61, P < 0.001) with the change in proteinuria. No other parameter correlated with the improvement in renal function. CONCLUSIONS: Blockade of the renin-angiotensin system in type 2 diabetic patients with diabetic nephropathy reduces uMCP-1 levels and improves renal function. Because MCP-1 induces monocyte immigration and differentiation to macrophages, which augment extracellular matrix production and tubulointerstitial fibrosis, pharmacological reduction of angiotensin II may also exert its beneficial effects in diabetic nephropathy by downregulation of renal MCP-1. PMID- 12882874 TI - Quantitative insulin sensitivity check index and the reciprocal index of homeostasis model assessment in normal range weight and moderately obese type 2 diabetic patients. AB - OBJECTIVE: To investigate whether the quantitative insulin sensitivity check index (QUICKI) and the reciprocal index of homeostasis model assessment (1/HOMA IR) derived from fasting plasma glucose and insulin level are excellent surrogate indices of insulin resistance in both normal range-weight and moderately obese type 2 diabetic and healthy subjects. RESEARCH DESIGN AND METHODS: The association between QUICKI or 1/HOMA-IR and insulin resistance index assessed by euglycemic-hyperinsulinemic clamp (clamp-IR) was investigated in 121 type 2 diabetic and 29 healthy subjects recruited from among 120 (age 55 +/- 11, 48 +/- 15, and 52 +/- 15 years [means +/- SD], respectively). Type 2 diabetic subjects were divided into groups of 76 normal range-weight and 45 moderately obese subjects (BMI 21.4 +/- 2.3 vs. 27.2 +/- 2.2 kg/m(2), P < 0.0001). RESULTS: QUICKI and 1/HOMA-IR were significantly lower in the moderately obese group than in the normal range-weight type 2 diabetic and healthy groups (n = 120) (QUICKI, 0.338 +/- 0.030, 0.371 +/- 0.037, and 0.389 +/- 0.041, respectively, P < 0.0001; 1/HOMA IR, 0.50 +/- 0.33, 0.92 +/- 0.55, and 1.24 +/- 0.82, P < 0.0001). QUICKI was strongly correlated with clamp-IR in normal range-weight, moderately obese type 2 diabetic, and healthy subjects (r = 0.641, 0.570, and 0.502, respectively; all subjects, r = 0.608, P < 0.01) and 1/HOMA-IR exhibited correlations comparable to those of QUICKI with clamp-IR (r = 0.637, 0.530, and 0.461, respectively; all subjects, r = 0.589, P < 0.001). In multiple regression models including QUICKI or 1/HOMA-IR as an independent variable, the estimation formula accounted for 55% of the variability of clamp-IR for the group of all type 2 diabetic subjects (R(2) = 0.547 and 0.551, respectively, P 41% reductions in skeletal muscle and white adipose tissue glucose uptake, but without alteration of hepatic insulin action. Munc18c transgenic mice exhibited approximately 40% decreases in whole-body glycogen/lipid synthesis, skeletal muscle glycogen synthesis, and glycolysis. Glucose intolerance in Munc18c transgenic mice was reversed by repression of transgene expression using tetracycline or by simultaneous overexpression of Syn4 protein. In addition, Munc18c transgenic mice had depressed serum insulin levels, reflecting a threefold reduction in insulin secretion from islets isolated therefrom, thus uncovering roles for Munc18c and/or Syn4 in insulin granule exocytosis. Taken together, these results indicate that balance, more than absolute abundance, of Munc18c and Syn4 proteins directly affects whole-body glucose homeostasis through alterations in insulin secretion and insulin action. PMID- 12882906 TI - A peroxovanadium compound stimulates muscle glucose transport as powerfully as insulin and contractions combined. AB - Stimulation of glucose transport by insulin involves tyrosine phosphorylation of the insulin receptor (IR) and IR substrates (IRSs). Peroxovanadates inhibit tyrosine phosphatases, also resulting in tyrosine phosphorylation of the IRSs. Muscle contractions stimulate glucose transport by a mechanism independent of the insulin-signaling pathway. We found that the peroxovanadate compound bis peroxovanadium,1,10-phenanthrolene [bpV(phen)] stimulates glucose transport to the same extent as the additive effects of maximal insulin and contraction stimuli. Translocation of GLUT4 to the cell surface mediates stimulation of glucose transport. There is evidence suggesting there are separate insulin- and contraction-stimulated pools of GLUT4-containing vesicles. We tested the hypothesis that bpV(phen) stimulates both the insulin- and the contraction activated pathways. Stimulation of glucose transport and GLUT4 translocation by bpV(phen) was completely blocked by the phosphatidylinositol 3-kinase (PI 3-K) inhibitors wortmannin and LY294002. The combined effect of bpV(phen) and contractions was no greater than that of bpV(phen) alone. Activation of the IRS PI 3-K signaling pathway was much greater with bpV(phen) than with insulin. Our results suggest that the GLUT4 vesicles that are normally translocated in response to contractions but not insulin can respond to the signal generated via the IRS-PI 3-K pathway if it is sufficiently powerful. PMID- 12882907 TI - Activation of protein kinase C-zeta by insulin and phosphatidylinositol-3,4,5 (PO4)3 is defective in muscle in type 2 diabetes and impaired glucose tolerance: amelioration by rosiglitazone and exercise. AB - Insulin resistance in type 2 diabetes is partly due to impaired glucose transport in skeletal muscle. Atypical protein kinase C (aPKC) and protein kinase B (PKB), operating downstream of phosphatidylinositol (PI) 3-kinase and its lipid product, PI-3,4,5-(PO(4))(3) (PIP(3)), apparently mediate insulin effects on glucose transport. We examined these signaling factors during hyperinsulinemic-euglycemic clamp studies in nondiabetic subjects, subjects with impaired glucose tolerance (IGT), and type 2 diabetic subjects. In nondiabetic control subjects, insulin provoked twofold increases in muscle aPKC activity. In both IGT and diabetes, aPKC activation was markedly (70-80%) diminished, most likely reflecting impaired activation of insulin receptor substrate (IRS)-1-dependent PI 3-kinase and decreased ability of PIP(3) to directly activate aPKCs; additionally, muscle PKC zeta levels were diminished by 40%. PKB activation was diminished in patients with IGT but not significantly in diabetic patients. The insulin sensitizer rosiglitazone improved insulin-stimulated IRS-1-dependent PI 3-kinase and aPKC activation, as well as glucose disposal rates. Bicycle exercise, which activates aPKCs and stimulates glucose transport independently of PI 3-kinase, activated aPKCs comparably to insulin in nondiabetic subjects and better than insulin in diabetic patients. Defective aPKC activation contributes to skeletal muscle insulin resistance in IGT and type 2 diabetes, rosiglitazone improves insulin stimulated aPKC activation, and exercise directly activates aPKCs in diabetic muscle. PMID- 12882908 TI - Insulin-stimulated protein kinase C lambda/zeta activity is reduced in skeletal muscle of humans with obesity and type 2 diabetes: reversal with weight reduction. AB - In humans with obesity or type 2 diabetes, insulin target tissues are resistant to many actions of insulin. The atypical protein kinase C (PKC) isoforms lambda and zeta are downstream of phosphatidylinositol-3 kinase (PI3K) and are required for maximal insulin stimulation of glucose uptake. Phosphoinositide-dependent protein kinase-1 (PDK-1), also downstream of PI3K, mediates activation of atypical PKC isoforms and Akt. To determine whether impaired PKClambda/zeta or PDK-1 activation plays a role in the pathogenesis of insulin resistance, we measured the activities of PKClambda/zeta and PDK-1 in vastus lateralis muscle of lean, obese, and obese/type 2 diabetic humans. Biopsies were taken after an overnight fast and after a 3-h hyperinsulinemic-euglycemic clamp. Obese subjects were also studied after weight loss on a very-low-calorie diet. Insulin stimulated glucose disposal rate is reduced 26% in obese subjects and 62% in diabetic subjects (both comparisons P < 0.001). Insulin-stimulated insulin receptor substrate (IRS)-1 tyrosine phosphorylation and PI3K activity are impaired 40-50% in diabetic subjects compared with lean or obese subjects. Insulin stimulates PKClambda/zeta activity approximately 2.3-fold in lean subjects; the increment above basal is reduced 57% in obese and 65% in diabetic subjects. PKClambda/zeta protein amount is decreased 46% in diabetic subjects but is normal in obese nondiabetic subjects, indicating impaired insulin action on PKClambda/zeta. Importantly, weight loss in obese subjects normalizes PKClambda/zeta activation and increases IRS-1 phosphorylation and PI3K activity. Insulin also stimulates PDK-1 activity approximately twofold with no impairment in obese or diabetic subjects. In contrast to our previous data on Akt, reduced insulin-stimulated PKClambda/zeta activity could play a role in the pathogenesis of insulin resistance in muscle of obese and type 2 diabetic subjects. PMID- 12882909 TI - Rosiglitazone improves downstream insulin receptor signaling in type 2 diabetic patients. AB - Thiazolidinediones (TZDs) improve glycemic control and insulin sensitivity in patients with type 2 diabetes. To determine whether the TZD-induced improvement in glycemic control is associated with enhanced insulin receptor signaling in skeletal muscle, 20 type 2 diabetic patients received a 75-g oral glucose tolerance test (OGTT) and euglycemic insulin (80 mU x m(-2) x min(-1)) clamp with [3-(3)H]glucose/indirect calorimetry/vastus lateralis muscle biopsies before and after 16 weeks of rosiglitazone treatment. Six age-matched nondiabetic subjects served as control subjects. RSG improved fasting plasma glucose (185 +/- 8 to 139 +/- 5 mg/dl), mean plasma glucose during the OGTT (290 +/- 9 to 225 +/- 6 mg/dl), HbA(1c) (8.5 +/- 0.3 to 7.1 +/- 0.3%), insulin-mediated total-body glucose disposal (TGD) (6.9 +/- 0.7 to 9.2 +/- 0.8 mg x kg(-1) fat-free mass x min(-1)) (all P < 0.001), and decreased fasting plasma free fatty acid (FFA) (789 +/- 59 to 656 +/- 50 micro Eq/l) and mean FFA during the OGTT (644 +/- 41 to 471 +/- 35 micro Eq/l) (both P < 0.01). Before RSG treatment, insulin infusion did not significantly increase insulin receptor tyrosine phosphorylation (0.95 +/- 0.10 to 1.08 +/- 0.13 density units; NS) but had a small stimulatory effect on insulin receptor substrate (IRS)-1 tyrosine phosphorylation (1.05 +/- 0.10 to 1.21 +/- 0.12 density units; P < 0.01) and the association of p85 with IRS-1 (0.94 +/- 0.06 to 1.08 +/- 0.06 activity units; P < 0.01). RSG therapy had no effect on basal or insulin-stimulated insulin receptor tyrosine phosphorylation but increased insulin stimulation of IRS-1 tyrosine phosphorylation (1.13 +/- 0.11 to 1.56 +/- 0.17 density units; P < 0.01 vs. prerosiglitazone) and p85 association with IRS-1 (1.00 +/- 0.06 to 1.27 +/- 0.07 activity units; P < 0.05 vs. prerosiglitazone). In control and type 2 diabetic subjects, TGD/nonoxidative glucose disposal correlated positively with the insulin-stimulated increments in IRS-1 tyrosine phosphorylation (r = 0.52/r = 0.57, P < 0.01) and inversely with the plasma FFA concentration during the insulin clamp (r = -0.55/r = -0.53, P < 0.01). However, no significant association between plasma FFA concentrations during the insulin clamp and the increment in either IRS-1 tyrosine phosphorylation or the association of p85 with IRS-1 was observed. In conclusion, in type 2 diabetic patients, rosiglitazone treatment enhances downstream insulin receptor signaling in muscle and decreases plasma FFA concentration while improving glycemic control. PMID- 12882910 TI - Central pro-opiomelanocortin gene delivery results in hypophagia, reduced visceral adiposity, and improved insulin sensitivity in genetically obese Zucker rats. AB - Zucker (fa/fa) rats with defective leptin receptors are obese, hyperphagic, and hyperinsulinemic. For testing whether chronic activation of the central melanocortin pathway can bypass the defective leptin signaling and normalize altered energy homeostasis in these rats, recombinant adeno-associated virus encoding pro-opiomelanocortin (rAAV-POMC) or control vector was delivered bilaterally into the basal hypothalamus with coordinates targeting the arcuate nucleus. Thirty-eight days after POMC gene delivery, hypothalamic POMC expression increased fourfold and melanocortin signaling (indicated by phosphorylation of CREB) increased by 62% with respect to controls. There was a sustained reduction in food intake, a moderate but significant attenuation of weight gain, and a 24% decrease in visceral adiposity in rAAV-POMC rats. POMC gene delivery enhanced uncoupling protein 1 in brown adipose tissue (BAT) by more than fourfold. Fasting serum leptin, insulin, and cholesterol levels were also significantly reduced by rAAV-POMC treatment. This study demonstrates that targeted POMC gene delivery in the hypothalamus suppresses food intake and weight gain and reduces visceral adiposity and hyperinsulinemia in leptin-resistant obese Zucker rats. The mechanisms may involve the sustained hypophagia and the augmentation of thermogenesis in BAT. PMID- 12882911 TI - Variation in type 2 diabetes--related traits in mouse strains susceptible to diet induced obesity. AB - C57BL/6J (B6) and AKR/J (AKR) inbred strains of mice develop a comparable degree of obesity when fed a high-fat diet. However, although obese B6 mice are more glucose intolerant, obese AKR mice are more insulin resistant. To understand the basis for these strain differences, we characterized features of adiposity and glucose homeostasis in mice fed a high-fat diet for 8 weeks. The results indicated that despite hyperglycemia and impaired glucose tolerance, B6 mice have lower plasma insulin and are more insulin sensitive than AKR mice. Compared with adipose tissue of AKR mice, adipose tissue of B6 mice contained about threefold higher levels of total membrane-bound GLUT4 protein, whereas in skeletal muscle the levels were similar. Uptake of 2-[(14)C]deoxyglucose in vivo was reduced by a high-fat diet in adipose tissue, but not in skeletal muscle. Surprisingly, no significant differences in uptake occurred between the strains, despite the differences in GLUT4; however, glucose flux was calculated to be slightly higher in B6 mice. Higher expression of PEPCK in the liver of B6 mice, under both standard-diet and high-fat-diet conditions, suggests a plausible mechanism for elevated glycemia in these mice. In conclusion, phenotypic variation in insulin resistance and glucose production in the B6 and AKR strains could provide a genetic system for the identification of genes controlling glucose homeostasis. PMID- 12882912 TI - Blockade of tumor necrosis factor-related apoptosis-inducing ligand exacerbates type 1 diabetes in NOD mice. AB - Tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) is expressed in different tissues and cells, including pancreas and lymphocytes, and can induce apoptosis in various tumor cells but not in most normal cells. The specific roles of TRAIL in health and disease remain unclear. Here we show by cDNA array analyses that TRAIL gene expression is upregulated in pancreatic islets during the development of autoimmune type 1 diabetes in nonobese diabetic (NOD) mice and in Min6 islet beta-cells activated by TNF-alpha + interferon gamma. However, stimulation of freshly isolated pancreatic islets or Min6 cells with TRAIL did not induce their apoptosis. TRAIL blockade exacerbates the onset of type 1 diabetes in NOD.Scid recipients of transferred diabetogenic T-cells and in cyclophosphamide-treated NOD mice. TRAIL inhibits the proliferation of NOD diabetogenic T-cells by suppressing interleukin (IL)-2 production and cell cycle progression, and this inhibition can be rescued in the presence of exogenous IL 2. cDNA array and Western blot analyses indicate that TRAIL upregulates the expression of the cdk inhibitor p27(kip1). Our data suggest that TRAIL is an important immune regulator of the development of type 1 diabetes. PMID- 12882913 TI - Prevention of diabetes in NOD mice by administration of dendritic cells deficient in nuclear transcription factor-kappaB activity. AB - Abnormalities of dendritic cells (DCs) have been identified in type 1 diabetic patients and in nonobese diabetic (NOD) mice that are associated with augmented nuclear transcription factor (NF)-kappaB activity. An imbalance that favors development of the immunogenic DCs may predispose to the disease, and restoration of the balance by administration of DCs deficient in NF-kappaB activity may prevent diabetes. DCs propagated from NOD mouse bone marrow and treated with NF kappaB-specific oligodeoxyribonucleotide (ODN) in vitro (NF-kappaB ODN DC) were assessed for efficacy in prevention of diabetes development in vivo. Gel shift assay with DC nuclear extracts confirmed specific inhibition of NF-kappaB DNA binding by NF-kappaB ODN. The costimulatory molecule expression, interleukin (IL) 12 production, and immunostimulatory capacity in presenting allo- and islet associated antigens by NF-kappaB ODN DC were significantly suppressed. NF-kappaB ODN renders DCs resistant to lipopolysaccharide stimulation. Administration of 2 x 10(6) NF-kappaB ODN DCs into NOD mice aged 6-7 weeks effectively prevented the onset of diabetes. T-cells from pancreatic lymph nodes of NF-kappaB ODN DC treated animals exhibited hyporesponsiveness to islet antigens with low production of interferon-gamma and IL-2. These findings provide novel insights into the mechanisms of autoimmune diabetes and may lead to development of novel preventive strategies. PMID- 12882914 TI - Endothelial differentiation gene receptors in pancreatic islets and INS-1 cells. AB - The endothelial differentiation gene (EDG) receptors are a class of G protein coupled receptors. EDG-1, -3, -5, -6, and -8 bind the bioactive lipid sphingosine 1-phosphate (SPP) as the primary signaling ligand. EDG-2, -4, and -7 bind the ligand lysophosphatidic acid. EDG-1, -2, -3, -5, -6, and -7, but not -8, mRNAs were expressed in isolated rat pancreatic islets, whereas INS-1 insulinoma cells expressed only EDG-1, -2, -3, and -5 mRNAs. EDG-4 mRNA was expressed in mouse islets. EDG-1 mRNA but not EDG-3 mRNA was rapidly induced relative to 18S rRNA after stimulation of isolated islets with phorbol 12-myristate 13-acetate (PMA) or cholecystokinin-8S for 2 h. The protein kinase C inhibitor GF 109203X blocked the EDG-1 induction by PMA. Similarly, in islets stimulated for 2 h with 17 mmol/l glucose, the relative EDG-1 mRNA levels increased almost twofold compared with levels in control islets at 5.5 mmol/l glucose. In contrast, after 11 mmol/l glucose stimulation for 7 days, the relative levels of rat islet EDG-1 mRNA were significantly reduced to 54% below that of islets cultured at 5.5 mmol/l glucose. There was no change in relative EDG-3 mRNA levels. Stimulation of EDG receptors in islets and INS-1 cells with SPP inhibited glucagon-like peptide 1 (GLP-1) stimulated cAMP production and insulin secretion in a concentration-dependent manner. Pertussis toxin antagonized the SPP effects on insulin release. Thus, EDG receptors are expressed in pancreatic islet beta-cells and G(i) seems to mediate the inhibition by SPP of adenylyl cyclase and cAMP formation and inhibition of the stimulation of insulin secretion by GLP-1. PMID- 12882915 TI - Free radicals and the pathogenesis of type 1 diabetes: beta-cell cytokine mediated free radical generation via cyclooxygenase-2. AB - Free radical formation evoked by proinflammatory cytokines has been suggested to be involved in the destruction of beta-cells in the course of type 1 diabetes development. However, there is no direct evidence to support this hypothesis. In this study, we used electron paramagnetic resonance spectroscopy in conjunction with spin-trapping methodology to directly determine whether cytokines give rise to free radical formation in the islets. Our results demonstrate that direct, in vivo administration of tumor necrosis factor-alpha (1,000 units), interleukin 1beta (1,000 units), and interferon-gamma (2,000 units) into the rat pancreas through a bile duct cannula leads to the formation of lipid-derived free radicals in this tissue. These free radicals most likely are generated by the beta-cells because previous depletion of these cells by streptozotocin abolished the cytokine-induced free radical formation. Furthermore, macrophage depletion was found to decrease the production of free radicals. Inhibition of the enzyme inducible cyclooxygenase (COX-2) and the transcription factor nuclear factor kappaB (NF-kappaB) significantly diminished the free radicals' signal intensity, implicating these factors in the formation of free radicals. We have also demonstrated that cytokine treatment leads to the activation of NF-kappaB in the pancreatic islets of the rats. PMID- 12882916 TI - Small-conductance calcium-activated K+ channels are expressed in pancreatic islets and regulate glucose responses. AB - Glucose-stimulated insulin secretion is associated with transients of intracellular Ca(2+) concentration [Ca(2+)](i) in the pancreatic beta-cell. We identified the expression and function of specific small-conductance Ca(2+) activated K(+) (SK) channel genes in insulin-secreting cells. The presence of mRNA for SK1, -2, -3, and -4 (intermediate-conductance Ca(2+)-activated K(+) 1 [IK1]) channels was demonstrated by RT-PCR in rodent islets and insulinoma cells. SK2 and -3 proteins in mouse islets were detected by immunoblot and immunocytochemistry. In the tTA-SK3 tet-off mouse, a normal amount of SK3 protein was present in islets, but it became undetectable after exposure to doxycycline (DOX), which inhibits the transcription of the tTA-SK3 gene. The SK/IK channel blockers apamin, dequalinium, and charybdotoxin caused increases in average [Ca(2+)](i) levels and in frequency of [Ca(2+)](i) oscillations in wild-type mouse islets. In SK3-tTA tet-off mice, the addition of apamin with glucose and tetraethylammonium (TEA) caused a similar elevation in [Ca(2+)](i), which was greatly diminished after DOX suppression of SK3 expression. We conclude that SK1, -2, -3, and IK1 (SK4) are expressed in islet cells and insulin-secreting cells and are able to influence glucose-induced calcium responses, thereby regulating insulin secretion. PMID- 12882917 TI - Characterization of endocrine progenitor cells and critical factors for their differentiation in human adult pancreatic cell culture. AB - We have reproduced a previously described method for the in vitro generation of endocrine cells in adult human pancreatic tissue culture. The aim of this study was to characterize the nature of pancreatic progenitor cells and to identify the factors necessary for their differentiation in this model. During monolayer expansion, two types of cells proliferated sequentially; first cytokeratin 19 (CK19)-positive ductal epithelial cells and then nestin-positive fibroblastoid cells. After the bromodeoxyuridine-labeled cells were traced in differentiated islet buds, some of the proliferating ductal cells had differentiated into endocrine cells, whereas nestin-positive cells could not give rise to endocrine tissue. Serum-free culture was found to be an absolute requirement for the endocrine differentiation to occur. Also, overlay of the cells with Matrigel was essential, whereas nicotinamide had a potentiating effect. The in vitro-generated islet buds released insulin in response to glucose nearly as efficiently as native islets. When transplanted under the kidney capsule of nude mice, only one of five grafts demonstrated further growth with foci of both endocrine and exocrine differentiation. Our results support the previous notion that pancreatic progenitor cells represent a subpopulation of ductal epithelial cells. No evidence was found for the development of endocrine cells from nestin-positive stem cells. PMID- 12882918 TI - Pancreatic precursors and differentiated islet cell types from murine embryonic stem cells: an in vitro model to study islet differentiation. AB - Embryonic stem (ES) cells differentiating in vitro reproduce many facets of early embryonic development, including the expression of developmentally regulated transcription factors and the differentiation of multipotent precursor cells. ES cells were evaluated for their ability to differentiate into pancreatic and islet lineage-restricted stages including pancreatic duodenal homeobox 1 (PDX1) positive pancreatic precursor cells, early endocrine cell progenitors, and islet hormone-producing cells. Following growth and differentiation in nonselective medium containing serum, murine ES cells spontaneously differentiated into cells individually expressing each of the four major islet hormones: insulin, glucagon, somatostatin, and pancreatic polypeptide. PDX1 immunostaining cells appeared first, before hormone-positive cells had emerged. Hormone-positive cells appeared within focal clusters of cells coexpressing PDX1 and the nonclassical hormone markers peptide YY (YY) and islet amyloid polypeptide (IAPP) in combination with the definitive hormones, characteristic of endocrine cells appearing during early pancreaticogenesis. This system allows the investigation of many facets of islet development since it promotes the appearance of the complete range of islet phenotypes and reproduces important developmental stages of normal islet cytodifferentiation in differentiating ES cell cultures. PMID- 12882919 TI - Diabetogenic potential of human pathogens uncovered in experimentally permissive beta-cells. AB - Pancreatic beta-cell antiviral defense plays a critical role in protection from coxsackievirus B4 (CVB4)-induced diabetes. In the present study, we tested the hypothesis that interferon (IFN)-induced antiviral defense determines beta-cell survival after infection by the human pathogen CVB3, cytomegalovirus (CMV), and lymphocytic choriomeningitis virus (LCMV). We demonstrated that mice harboring beta-cells that do not respond to IFN because of the expression of the suppressor of cytokine signaling-1 (SOCS-1) succumb to an acute form of type 1 diabetes after infection with CVB3. Interestingly, the tropism of the virus was altered in SOCS-1 transgenic (Tg) mice, and CVB3 was detected in islet cells of SOCS-1-Tg mice before beta-cell loss and the onset of diabetes. Furthermore, insulitis was increased in SOCS-1-Tg mice after infection with murine CMV, and a minority of the mice developed overt diabetes. However, infection with LCMV failed to cause beta-cell destruction in SOCS-1 Tg mice. These findings suggest that CVB3 can cause diabetes in a host lacking adequate beta-cell antiviral defense, and that incomplete target cell antiviral defense may enhance susceptibility to diabetes triggered by CMV. In conclusion, suppressed beta-cell antiviral defense reveals the diabetogenic potential of two pathogens previously linked to the onset of type 1 diabetes in humans. PMID- 12882920 TI - Label-retaining cells in the rat pancreas: location and differentiation potential in vitro. AB - Islets of Langerhans are micro-organs scattered throughout the pancreas that contain insulin-producing cells, called beta-cells. Although new light has been recently shed on beta-cell development, information on the phenotype and location of beta-stem cells remains scarce. Here, we provide evidence that beta-stem cells are slow-cycling cells located within and around the islets of Langerhans. First, using a bromodeoxyuridine (BrdU) pulse/chase approach, we detected BrdU-retaining cells in vivo in the islet area of rat pancreata. These cells were negative for endocrine markers but expressed Pdx1, a marker for pancreatic stem cells. Next, using an in vitro model that mimicked endocrine cell development, we found that BrdU-retaining cells were capable of differentiating into beta-cells. Taken together, these observations demonstrate that BrdU retention is a property of beta-stem cells. PMID- 12882921 TI - K(ATP) channels and pancreatic islet blood flow in anesthetized rats: increased blood flow induced by potassium channel openers. AB - K(ATP) channels are important for insulin secretion and depolarization of vascular smooth muscle. In view of the importance of drugs affecting K(ATP) channels in the treatment of diabetes, we investigated the effects of these channels on splanchnic blood perfusion in general and pancreatic islet blood flow in particular. We treated anesthetized Sprague-Dawley rats with the K(ATP) channel openers diazoxide or NNC 55-0118 or the K(ATP) channel closer glipizide. Both diazoxide and NNC 55-0118 dose-dependently increased total pancreatic and islet blood flow in the presence of moderate hyperglycemia, but had no effects on the blood perfusion of other splanchnic organs. Diazoxide markedly lowered the mean arterial blood pressure and thus increased vascular conductance in all organs studied. NNC 55-0118 had much smaller effects on the blood pressure. Glipizide did not affect total pancreatic blood flow, but decreased islet blood flow by 50% in the presence of hypoglycemia. We conclude that K(ATP) channels actively participate in the blood flow regulation of the pancreatic islets and that substances affecting such channels may also influence islet blood flow. PMID- 12882922 TI - Insulin constitutively secreted by beta-cells is necessary for glucose-stimulated insulin secretion. AB - Four hypotheses have been posited on the role of insulin in glucose-stimulated insulin secretion; available evidence has supported insulin as being 1) essential, 2) a positive modulator, 3) a negative modulator, or 4) not necessary. Because circulating insulin levels in mice, before or after intraperitoneal glucose injection, are sufficient to elicit insulin responses in insulin sensitive tissues, it is likely that beta-cell insulin receptors are continuously exposed to stimulating concentrations of insulin. To determine whether constitutively secreted insulin is necessary for glucose-stimulated insulin secretion, CD1 male mouse islets were incubated for 30 min at 4 degrees C in the absence (control) or presence of anti-insulin (1 micro g/ml) or anti-IgG (1 micro g/ml). Then islets were exposed to 3, 11, or 25 mmol/l glucose or to 20 mmol/l arginine. Nontreated islets exhibited first- and second-phase glucose-stimulated insulin secretion. Control and anti-IgG-treated islets, after a 5-min lag phase, increased their insulin secretion in 25 mmol/l glucose. Anti-insulin-treated islets secreted insulin at a basal rate in 3 or 25 mmol/l glucose buffers. Insulin secretion stimulated by 20 mmol/l arginine was the same in islets pretreated with either antibody and showed no lag phase. Taken together, these data suggest that constitutively secreted insulin is required and sufficient for beta-cells to maintain sensitivity to glucose. PMID- 12882923 TI - Pancreatic beta-cell lipotoxicity induced by overexpression of hormone-sensitive lipase. AB - Lipid perturbations associated with triglyceride overstorage in beta-cells impair insulin secretion, a process termed lipotoxicity. To assess the role of hormone sensitive lipase, which is expressed and enzymatically active in beta-cells, in the development of lipotoxicity, we generated transgenic mice overexpressing hormone-sensitive lipase specifically in beta-cells. Transgenic mice developed glucose intolerance and severely blunted glucose-stimulated insulin secretion when challenged with a high-fat diet. As expected, both lipase activity and forskolin-stimulated lipolysis was increased in transgenic compared with wild type islets. This was reflected in significantly lower triglycerides levels in transgenic compared with wild-type islets in mice receiving the high-fat diet, whereas no difference in islet triglycerides was found between the two genotypes under low-fat diet conditions. Our results highlight the importance of mobilization of the islet triglyceride pool in the development of beta-cell lipotoxicity. We propose that hormone-sensitive lipase is involved in mediating beta-cell lipotoxicity by providing ligands for peroxisome proliferator-activated receptors and other lipid-activated transcription factors, which in turn alter the expression of critical genes. One such gene might be uncoupling protein-2, which was found to be upregulated in transgenic islets, a change that was accompanied by decreased ATP levels. PMID- 12882924 TI - Vesicular inhibitory amino acid transporter is present in glucagon-containing secretory granules in alphaTC6 cells, mouse clonal alpha-cells, and alpha-cells of islets of Langerhans. AB - Islets of Langerhans contain gamma-aminobutyrate (GABA) and may use it as an intercellular transmitter. In beta-cells, GABA is stored in synaptic-like microvesicles and secreted through Ca(2+)-dependent exocytosis. Vesicular inhibitory amino acid transporter (VIAAT), which is responsible for the storage of GABA and glycine in neuronal synaptic vesicles, is believed to be responsible for the storage and secretion of GABA in beta-cells. However, a recent study by Chessler et al. indicated that VIAAT is expressed in the mantle region of islets. In the present study, we investigated the precise localization of VIAAT in rat islets of Langerhans and clonal islet cells and found that it is present in alpha cells, a minor population of F-cells and alphaTC6 cells, and clonal alpha-cells but not in beta-cells, delta-cells, or MIN6 m9-cells (clonal beta-cells). Combined biochemical, immunohistochemical, and electronmicroscopical evidence indicated that VIAAT is specifically localized with glucagon-containing secretory granules in alpha-cells. ATP-dependent uptake of radiolabeled GABA, which is energetically coupled with a vacuolar proton pump, was detected in digitonin permeabilized alphaTC6 cells as well as in MIN6 m9 cells. These results demonstrate that functional neuronal VIAAT is present in glucagon-containing secretory granules in alpha-cells and suggest that the ATP-dependent GABA transporter in beta-cells is at least immunologically distinct from VIAAT. Because glucagon-containing secretory granules also contain vesicular glutamate transporter and store L-glutamate, as demonstrated by Hayashi et al., the present results suggest more complex features of the GABAergic phenotype of islets than previously supposed. PMID- 12882925 TI - Vascular endothelial function and blood pressure homeostasis in mice overexpressing IGF binding protein-1. AB - IGFs and their binding proteins (IGFBPs) play a significant role in metabolic regulation, and there is growing evidence that they also exert important vascular effects. IGFBP-1 contributes to glucose counterregulation, and observational studies demonstrate an inverse association between circulating IGFBP-1 levels and cardiovascular risk factors. Furthermore, IGFBP-1 levels are lower in subjects with overt macrovascular disease. We therefore hypothesized that IGFBP-1 exerts potentially beneficial effects, either directly or indirectly, on blood pressure regulation and vascular function. We tested this hypothesis using a unique transgenic mouse, which overexpresses human IGFBP-1, and explored the effect of this protein on metabolic, blood pressure, and vascular homeostasis. IGFBP-1 overexpressing mice exhibited postprandial hyperinsulinemia with preservation of glucocompetence and insulin sensitivity. Blood pressure was unchanged in the fasting state but was significantly lower in transgenic mice after a carbohydrate load. Aortic rings from IGFBP-1-overexpressing mice were hypocontractile in response to vasoconstrictors, and relaxation responses were unimpaired. Basal nitric oxide production was increased and endothelial nitric oxide synthase mRNA expression upregulated in aortae of these mice. Our data suggest that IGFBP-1 plays an important and potentially beneficial role in regulating metabolic and vascular homeostasis. PMID- 12882926 TI - Cortisol elevations comparable to those that occur during hypoglycemia do not cause hypoglycemia-associated autonomic failure. AB - The concept of hypoglycemia-associated autonomic failure (HAAF) in diabetes posits that recent antecedent iatrogenic hypoglycemia causes both defective glucose counterregulation (by reducing the epinephrine response in the setting of an absent glucagon response) and hypoglycemia unawareness (by reducing the autonomic-sympathetic neural and adrenomedullary response and the resulting neurogenic [autonomic] symptom responses) and thus causes a vicious cycle of recurrent hypoglycemia. To assess the suggestion that it is the cortisol response to antecedent hypoglycemia that mediates HAAF, we tested the hypothesis that plasma cortisol elevations during euglycemia that are comparable to those that occur during hypoglycemia reduce sympathoadrenal and neurogenic symptom responses to subsequent hypoglycemia. To do this, 12 healthy subjects were studied with hyperinsulinemic-stepped hypoglycemic clamps the day after saline or cortisol (1.3 +/- 0.2 micro g. kg(-1) x min(-1)) infusions from 0930 to 1200 and from 1330 to 1600. Compared with saline, antecedent cortisol elevations did not reduce the sympathoadrenal (e.g., final plasma epinephrine levels of 674 +/- 84 vs. 606 +/- 80 pg/ml and final plasma norepinephrine levels of 332 +/- 26 vs. 304 +/- 26 pg/ml) or neurogenic symptom (e.g., final scores of 9.3 +/- 1.1 vs. 13.2 +/- 1.3) responses to subsequent hypoglycemia. Thus, these data do not support the suggestion that cortisol mediates HAAF. PMID- 12882927 TI - Higher offspring birth weight predicts the metabolic syndrome in mothers but not fathers 8 years after delivery: the Pune Children's Study. AB - In Europid populations, low birth weight of offspring predicts insulin resistance in the mother and cardiovascular disease in both parents. We investigated the association between birth weight of offspring and obesity and cardiovascular risk in the parents of 477 8-year-old children born at the King Edward Memorial Hospital, Pune, India. Eight years after the birth of the child, mothers (33 years of age, n = 459) of heavier babies were taller and more obese (BMI, fat mass, and waist circumference, all P < 0.001) than mothers of lighter babies. Increasing offspring birth weight predicted higher homeostasis model assessment for insulin resistance (P < 0.01) and metabolic syndrome in mothers (P < 0.001) (adjusted for offspring sex and birth order, maternal age, and socioeconomic status) but not hyperglycemia. Fathers (39 years of age, n = 398) of heavier babies were taller and heavier, independent of maternal size (P < 0.01, both), but were not more insulin resistant. Unlike other reports, lower offspring birth weight did not predict insulin resistance in fathers. Thus, urban Indian parents have a higher risk of being obese 8 years after delivery of a heavier child. Mothers but not fathers of heavier babies also have a higher risk of being insulin resistant and developing the metabolic syndrome. Our findings highlight the need for a better understanding of the relation between fetal growth and future health before contemplating public health interventions to improve fetal growth. PMID- 12882928 TI - Inflammation-sensitive plasma proteins are associated with future weight gain. AB - Cross-sectional studies have associated obesity and other components of the so called metabolic syndrome with low-grade inflammation. The temporal and causal relations of this association have not been fully explored. This study explored whether elevated levels of inflammation-sensitive plasma proteins (ISPs) (fibrinogen, orosomucoid, alpha1-antitrypsin, haptoglobin, and ceruloplasmin) are associated with future weight gain. Five ISPs were measured in 2,821 nondiabetic healthy men (38-50 years of age) who were reexamined after a mean follow-up of 6.1 years. Future weight gain was studied in relation to the number of elevated ISPs (i.e., in the top quartile). The proportion with a large weight gain (75th percentile >/= 3.8 kg) was 21.0, 25.9, 26.8, and 28.3%, respectively, among men with none, one, two, and three or more ISPs in the top quartile (P for trend 0.0005). This relation remained significant after adjustments for weight at baseline, follow-up time, height (at baseline and follow-up), physical inactivity (at baseline and follow-up), smoking (at baseline and follow-up), high alcohol consumption, and insulin resistance. The relations were largely similar for all individual ISPs. Elevated ISP levels predict a large weight gain in middle-aged men. This relation could contribute to the relation between inflammation, the metabolic syndrome, and cardiovascular disease. PMID- 12882929 TI - Protein kinase Cbeta selective inhibitor LY333531 attenuates diabetic hyperalgesia through ameliorating cGMP level of dorsal root ganglion neurons. AB - Streptozocin (STZ)-induced diabetic rats show hyperalgesia that is partially attributed to altered protein kinase C (PKC) activity. Both attenuated neuronal nitric oxide synthase (nNOS)-cGMP system and tetrodotoxin-resistant (TTX-R) Na channels in dorsal root ganglion neurons may be involved in diabetic hyperalgesia. We examined whether PKCbeta inhibition ameliorates diabetic hyperalgesia and, if so, whether the effect is obtained through action on neurons by testing nociceptive threshold in normal and STZ-induced diabetic rats treated with or without PKCbeta-selective inhibitor LY333531 (LY) and by assessing the implication of LY in either nNOS-cGMP system or TTX-R Na channels of isolated dorsal root ganglion neurons. The decreased nociceptive threshold in diabetic rats was improved either after 4 weeks of LY treatment or with a single intradermal injection into the footpads. The treatment of LY for 6 weeks significantly decreased p-PKCbeta and ameliorated a decrease in cGMP content in dorsal root ganglia of diabetic rats. The latter effect was confirmed in ex vivo condition. The treatment with NO donor for 4 weeks also normalized both diabetic hyperalgesia and decreased cGMP content in dorsal root ganglions. The expressions of nNOS and TTX-R Na channels were not changed with LY treatment. These results suggest that LY is effective for treating diabetic hyperalgesia through ameliorating the decrease in the nNOS-cGMP system. PMID- 12882930 TI - Prevention of incipient diabetic nephropathy by high-dose thiamine and benfotiamine. AB - Accumulation of triosephosphates arising from high cytosolic glucose concentrations in hyperglycemia is the trigger for biochemical dysfunction leading to the development of diabetic nephropathy-a common complication of diabetes associated with a high risk of cardiovascular disease and mortality. Here we report that stimulation of the reductive pentosephosphate pathway by high dose therapy with thiamine and the thiamine monophosphate derivative benfotiamine countered the accumulation of triosephosphates in experimental diabetes and inhibited the development of incipient nephropathy. High-dose thiamine and benfotiamine therapy increased transketolase expression in renal glomeruli, increased the conversion of triosephosphates to ribose-5-phosphate, and strongly inhibited the development of microalbuminuria. This was associated with decreased activation of protein kinase C and decreased protein glycation and oxidative stress-three major pathways of biochemical dysfunction in hyperglycemia. Benfotiamine also inhibited diabetes-induced hyperfiltration. This was achieved without change in elevated plasma glucose concentration and glycated hemoglobin in the diabetic state. High-dose thiamine and benfotiamine therapy is a potential novel strategy for the prevention of clinical diabetic nephropathy. PMID- 12882931 TI - Leptin increases lipoprotein lipase secretion by macrophages: involvement of oxidative stress and protein kinase C. AB - Recent data suggest that plasma leptin may represent a cardiovascular risk factor in diabetic patients. To gain further insight into the role of leptin in atherogenesis associated with diabetes, we investigated in the present study the role of this hormone in the regulation of macrophage lipoprotein lipase (LPL), a proatherogenic cytokine overexpressed in patients with type 2 diabetes. Treatment of human macrophages with leptin (1-10 nmol/l) increased LPL expression, at both the mRNA and protein levels. Pretreatment of these cells with anti-leptin receptor (Ob-R) antibody, protein kinase C (PKC) inhibitors, calphostin C, and GF109203X, or the antioxidant N-acetylcysteine (NAC) blocked the effects of leptin. Similar results were observed in leptin-treated J774 macrophages. In these cells, leptin increased the membrane expression of conventional PKC isoforms and downregulation of endogenous PKC expression abolished the effects of leptin on macrophage LPL expression. In leptin-treated J774 cells, enhanced LPL synthetic rate and increased binding of nuclear proteins to the activated protein 1 (AP-1) consensus sequence of the LPL gene promoter were also observed. This latter effect was abrogated by GF109203X. Overall, these data demonstrate that binding of leptin at the macrophage cell surface increases, through oxidative stress- and PKC-dependent pathways, LPL expression. This effect appears to be exerted at the transcriptional level and to involve AP-1 activation. PMID- 12882932 TI - Insulin prevents depolarization of the mitochondrial inner membrane in sensory neurons of type 1 diabetic rats in the presence of sustained hyperglycemia. AB - Mitochondrial dysfunction has been proposed as a mediator of neurodegeneration in diabetes complications. The aim of this study was to determine whether deficits in insulin-dependent neurotrophic support contributed to depolarization of the mitochondrial membrane in sensory neurons of streptozocin (STZ)-induced diabetic rats. Whole cell fluorescent video imaging using rhodamine 123 (R123) was used to monitor mitochondrial inner membrane potential (deltapsi(m)). Treatment of cultured dorsal root ganglia (DRG) sensory neurons from normal adult rats for up to 1 day with 50 mmol/l glucose had no effect; however, 1.0 nmol/l insulin increased deltapsi(m) by 100% (P < 0.05). To determine the role of insulin in vivo, STZ-induced diabetic animals were treated with background insulin and the deltapsi(m) of DRG sensory neurons was analyzed. Insulin therapy in STZ-induced diabetic rats had no effect on raised glycated hemoglobin or sciatic nerve polyol levels, confirming that hyperglycemia was unaffected. However, insulin treatment significantly normalized diabetes-induced deficits in sensory and motor nerve conduction velocity (P < 0.05). In acutely isolated DRG sensory neurons from insulin-treated STZ animals, the diabetes-related depolarization of the deltapsi(m) was corrected (P < 0.05). The results demonstrate that loss of insulin-dependent neurotrophic support may contribute to mitochondrial membrane depolarization in sensory neurons in diabetic neuropathy. PMID- 12882933 TI - Production of N(epsilon)-(carboxymethyl)lysine is impaired in mice deficient in NADPH oxidase: a role for phagocyte-derived oxidants in the formation of advanced glycation end products during inflammation. AB - Advanced glycation end products (AGEs) derived from glucose are implicated in the pathogenesis of diabetic vascular disease. However, many lines of evidence suggest that other pathways also promote AGE formation. One potential mechanism involves oxidants produced by the NADPH oxidase of neutrophils, monocytes, and macrophages. In vitro studies have demonstrated that glycolaldehyde, a product of serine oxidation, reacts with proteins to form N(epsilon)-(carboxymethyl)lysine (CML), a chemically well-characterized AGE. We used mice deficient in phagocyte NADPH oxidase (gp91-phox(-/-)) to explore the role of oxidants in AGE production in isolated neutrophils and intact animals. Activated neutrophils harvested from wild-type mice generated CML on ribonuclease A (RNase A), a model protein, by a pathway that required L-serine. CML formation by gp91-phox(-/-) neutrophils was impaired, suggesting that oxidants produced by phagocyte NADPH oxidase contribute to the cellular formation of AGEs. To determine whether these observations are physiologically relevant, we used isotope-dilution gas chromatography/mass spectrometry to quantify levels of protein-bound CML in mice suffering from acute peritoneal inflammation. Phagocytes from the gp91-phox(-/-) mice contained much lower levels of CML than those from the wild-type mice. Therefore, oxidants generated by phagocyte NADPH oxidase may play a role in AGE formation in vivo by a glucose-independent pathway. PMID- 12882934 TI - Expression of constitutively active cGMP-dependent protein kinase prevents glucose stimulation of thrombospondin 1 expression and TGF-beta activity. AB - Hyperglycemia is a crucial factor in the development of diabetic nephropathy. We previously showed that high glucose upregulates thrombospondin 1 (TSP1)-dependent transforming growth factor (TGF)-beta activation by altering cGMP-dependent protein kinase (PKG) activity as a result of decreased nitric oxide signaling. In the present study, we showed that high glucose concentrations significantly reduced endogenous PKG activity. To further examine the mechanisms by which PKG regulates TSP1 expression and TSP1-dependent TGF-beta activation, we generated stably transfected rat mesangial cells (RMCs) with inducible expression tetracycline-induced gene expression of the catalytic domain of PKG. After tetracycline induction, the catalytic domain of PKG is expressed as a cGMP independent active kinase. Expression of the catalytic domain prevented high glucose-mediated increases in transcription of the TSP1 gene with no alteration in TSP1 mRNA stability. Glucose stimulation of TSP1 protein expression and TGF beta bioactivity were also downregulated. TGF-beta-dependent fibronectin and type IV collagen expression under high glucose conditions were significantly reduced upon catalytic domain expression in transfected RMCs. These results show that constitutively active PKG inhibits the fibrogenic potential of high glucose through repression of TSP1-dependent TGF-beta bioactivity, suggesting that gene transfer of the catalytic domain of PKG might provide a new strategy for treatment of diabetic renal fibrosis. PMID- 12882935 TI - Microarray analysis of gene expression in the kidneys of new- and post-onset diabetic NOD mice. AB - We profiled the expression of 5,760 clones from a kidney subtraction library in the kidneys of three groups of NOD mice: nondiabetic, new-onset, and long-term diabetic. A total of 27 genes had lower expression and 1 gene (Gpx3) had higher expression in the new-onset diabetic mice compared with nondiabetic control NOD mice (P < 0.001). Similarly, 19 of the above 27 genes and 7 additional genes had higher expression and the Gpx3 gene had lower expression in long-term diabetic mice compared with controls (P < 0.001). Interestingly, only three genes may be different between new-onset and long-term diabetic mice (P < 0.0004). These genes are from diverse functional groups, including oxidative phosphorylation, free radical neutralization, channels, pumps, lipid processing, transcription and translation machinery, protein trafficking, constitutive protein processing, and immune function. The majority of these genes fall into four signaling pathways: insulin, transforming growth factor-beta, tumor necrosis factor-alpha, and peroxisome proliferator-activated receptor. The most significant expression change was found for the stearoyl-coenzyme A desaturase 1 (SCD1) gene (P < 10( 7)). The lower expression levels of the SCD1 gene in both diabetic groups compared with controls were further confirmed by Northern blot analysis and immunohistochemistry. PMID- 12882936 TI - Prevalence and characteristics of the metabolic syndrome in the San Antonio Heart and Framingham Offspring Studies. AB - The metabolic syndrome may be a common phenotype increasing risk for type 2 diabetes and cardiovascular disease. We assessed the prevalence and characteristics of the metabolic syndrome among population-based samples of 3,224 white subjects attending Framingham Offspring Study (FOS) exam 5 (1991-1995) and 1,081 non-Hispanic white and 1,656 Mexican-American subjects attending the San Antonio Heart Study (SAHS) phase II follow-up exam (1992-1996). Subjects were approximately 50% women, aged 30-79 years, without diabetes, and classified with the metabolic syndrome according to criteria for obesity, dyslipidemia, hyperglycemia, and hypertension proposed by the Third Report of the National Cholesterol Education Program's Adult Treatment Panel (ATP III) or the World Health Organization (WHO). We used regression models to estimate rates across ethnic groups and to assess the association of the metabolic syndrome with insulin resistance and predicted 10-year coronary heart disease (CHD) risk. Among FOS white subjects, the age- and sex-adjusted prevalence of the metabolic syndrome was 24% by both ATP III and WHO criteria; among SAHS non-Hispanic white subjects, 23 and 21%, respectively; and among SAHS Mexican-American subjects, 31 and 30%. Rates were highest among Mexican-American women (ATP III, 33%) and lowest among white women (21%). Subjects with the metabolic syndrome by ATP III criteria had higher age-, sex-, and ethnicity-adjusted levels of fasting insulin (11.3 micro U/ml), homeostasis model assessment of insulin resistance (2.7), and predicted CHD risk (11.8%) than those without the syndrome (5.9 micro U/ml, 1.3, and 6.4%, respectively; all P = 0.0001); differences were similar using WHO criteria. We conclude that the metabolic syndrome typically affects 20-30% of middle-aged adults in the U.S. By any criteria, subjects with the metabolic syndrome are more insulin resistant and at increased predicted risk for CHD versus those without the metabolic syndrome. PMID- 12882937 TI - Minimal model-based insulin sensitivity has greater heritability and a different genetic basis than homeostasis model assessment or fasting insulin. AB - Insulin resistance is an important risk factor for development of type 2 diabetes as well as other chronic conditions, including hypertension, cardiovascular disease, and colon cancer. To find genes for insulin resistance it is necessary to assess insulin action in large populations. We have previously measured insulin action in a large cohort of subjects (Insulin Resistance and Atherosclerosis Study [IRAS] Family Study) using the minimal model approach. In this study, we compare sensitivity from the minimal model (insulin sensitivity index [S(I)]) with the measure of insulin resistance emanating from the homeostasis model assessment (HOMA) approach. The former measure emerges from the glycemic response to endogenous and exogenous insulin; the latter is based solely on fasting measures of glucose and insulin. A total of 112 pedigrees were represented, including 1,362 individuals with full phenotypic assessment. Heritability of S(I) was significantly greater than that for HOMA (0.310 vs. 0.163) and for fasting insulin (0.171), adjusted for age, sex, ethnicity, and BMI. In addition, correlation between S(I) and either HOMA or fasting insulin was only approximately 50% accounted for by genetic factors, with the remainder accounted for by environment. Thus S(I), a direct measure of insulin sensitivity, is determined more by genetic factors rather than measures such as HOMA, which reflect fasting insulin. PMID- 12882938 TI - Identification of epistatic interaction involved in obesity using the KK/Ta mouse as a Type 2 diabetes model: is Zn-alpha2 glycoprotein-1 a candidate gene for obesity? AB - The KK/Ta strain serves as a suitable polygenic mouse model for the common form of type 2 diabetes associated with obesity in humans. Recently, we reported the susceptibility loci contributing to type 2 diabetes and related phenotypes in KK/Ta mice. In this study, we focused on expression in the kidneys and liver of KK/Ta and BALB/c mice using differential display (DD) PCR. Zn-alpha(2) glycoprotein-1 (Azgp1) mRNA levels were increased in the kidneys and liver in KK/Ta mice, and sequence analysis revealed a missense mutation. We analyzed the relationship between this polymorphism and various phenotypes in 208 KK/Ta x (BALB/c x KK/Ta) F1 backcross mice. Statistical analysis revealed that Azgp1 and D17Mit218 exhibit a suggestive linkage to body weight (8 weeks) (logarithm of odds 2.3 and 2.9, respectively). Moderate gene-gene interactions were observed at these loci. Adiponectin mRNA levels in 3T3-L1 cells transfected with the expression pcDNA 3.1 vector containing Azgp1 coding sequence of KK/Ta mice were significantly higher than those of BALB/c mice. These results suggest that Azgp1 is a possible candidate gene for regulation of body weight, elucidation of polygenic inheritance, and age-dependent changes in the genetic control of obesity. PMID- 12882939 TI - Genetic modifiers of the age at diagnosis of diabetes (MODY3) in carriers of hepatocyte nuclear factor-1alpha mutations map to chromosomes 5p15, 9q22, and 14q24. AB - Mutations in hepatocyte nuclear factor (HNF)-1alpha (MODY3) account for the largest proportion of maturity-onset diabetes of the young (MODY) cases in the U.S. This form of diabetes is characterized by impaired insulin secretion in response to glucose, but wide variability exists in the severity of hyperglycemia and in the age at which it becomes clinically manifest. We have previously shown that the age at onset of diabetes in MODY3 families is influenced by familial factors (including modifying genes) and exposure to diabetes in utero. To identify genes influencing the onset of MODY3, we conducted a genome scan in 13 extended MODY families in which diabetes segregates with an HNF-1alpha mutation. Linkage with age at onset of diabetes was assessed by genetic variance component analysis using SOLAR. The locus with the strongest evidence of linkage was on chromosome 14q24 (D14S588; logarithm of odds [LOD] = 2.58, P = 0.0004). This location overlaps with IDDM11 and includes SEL1L, a negative regulator of the Notch pathway that may control islet development. Linkage evidence also supported loci on 5p15 (D5S817; LOD = 2.44, P = 0.0004) and 9q22 (D9S910; LOD = 2.02, P = 0.0018). The latter matches a region linked to 2-h insulin levels in Pima Indians. Less strong linkage evidence was observed at three other regions: chromosomes 3p24 (LOD = 1.44), 7q21 (1.20), and 16q23 (1.51). Our data are consistent with the existence of multiple loci that contribute to the expression of the MODY3 phenotype. Identification of these genes will offer new insights into the pathophysiology of MODY that may, in turn, increase our understanding of the cellular events underlying more common forms of diabetes. PMID- 12882941 TI - Invited commentary: is indoor mold exposure a risk factor for asthma? PMID- 12882940 TI - Symptoms of wheeze and persistent cough in the first year of life: associations with indoor allergens, air contaminants, and maternal history of asthma. AB - In a cohort of 849 infants with an asthmatic sibling, the authors examined the relations of exposure to allergens (dust mite, cockroach, cat, and dog), nitrogen dioxide, and mold with symptoms of wheeze and persistent cough in the first year of life (1998-2000). Among infants whose mothers had physician-diagnosed asthma, neither dust mite allergen nor dog allergen was associated with either symptom. Exposure to cockroach allergen (Bla g 1 at >or=2 U/g) modestly increased the risk for wheeze (odds ratio (OR) = 1.87, 95% confidence interval (CI): 0.94, 3.71), and exposure to cat allergen modestly decreased the risk (OR = 0.60, 95% CI: 0.35, 1.03). Among infants of mothers with no asthma history, exposure to gas stoves (OR = 1.50, 95% CI: 1.05, 2.15) and wood-burning stoves (OR = 2.09, 95% CI: 1.12, 3.91) increased the risk of persistent cough. Similarly, measured nitrogen dioxide concentration was associated with persistent cough (OR = 1.21, 95% CI: 1.05, 1.40). Persistent mold affected both infants of mothers with asthma (for wheeze, OR = 2.27, 95% CI: 1.27, 4.07; for cough, OR = 1.83, 95% CI: 1.04, 3.22) and infants of mothers without asthma (for cough, OR = 1.55, 95% CI: 1.04, 2.31). Reported exposure was confirmed by an association of measured fungi with wheeze (OR = 1.23, 95% CI: 1.01, 1.49). This appears to have been the first study to measure all of these home exposures (indoor allergens, nitrogen dioxide, fungi) and to prospectively measure the frequency of infant wheeze and persistent cough. PMID- 12882942 TI - Maternal herpesvirus infections and risk of acute lymphoblastic leukemia in the offspring. AB - A critical role for infection in the etiology of childhood leukemia has repeatedly been suggested. The authors undertook a case-control study nested within national maternity cohorts with altogether 7 million years of follow-up to assess the relative role of three maternal herpesvirus infections in childhood acute lymphoblastic leukemia (ALL). Offspring of 550,000 mothers in Finland and Iceland formed the joint study cohort that was followed up for cancer in the offspring before age 15 years during 1975-1997 through national cancer registries. For each index mother-case pair, three or four matched control mother control pairs were identified from national population registers. First-trimester sera were retrieved from mothers of 342 ALL and 61 other leukemia cases and from 1,216 control mothers and were tested for antibodies to cytomegalovirus, Epstein Barr virus (EBV), and human herpesvirus 6. Serum EBV DNA was also analyzed. Conditional logistic regression-based estimates of relative risk (odds ratio) adjusted for birth order and sibship size, and population attributable fractions, were calculated. Only EBV immunoglobulin M positivity in EBV-immunoglobulin-G positive mothers was associated with a highly significant increased risk of ALL in the offspring (adjusted odds ratio = 2.9, 95% confidence interval: 1.5, 5.8). Results indicate that reactivation of maternal EBV infection is probably associated with childhood ALL. PMID- 12882943 TI - Physical activity and colorectal cancer. AB - Physical activity has been inconsistently associated with rectal cancer despite the consistent association between physical activity and colon cancer. In this study, the authors evaluated the association between physical activity and rectal cancer using the same questionnaire used to evaluate the previously reported association with colon cancer. A population-based study of 952 incident cases of cancer in the rectum and rectosigmoid junction and 1,205 age- and sex-matched controls was conducted in Utah and northern California at the Kaiser Permanente Medical Care Program between 1997 and 2002. Vigorous physical activity was associated with reduced risk of rectal cancer in both men and women (odds ratio (OR) = 0.60, 95% confidence interval (CI): 0.44, 0.81 for men; OR = 0.59, 95% CI: 0.40, 0.86 for women). Among men, moderate levels of physical activity also were associated with reduced risk of rectal cancer (OR = 0.70, 95% CI: 0.51, 0.97). Participation in vigorous activity over the past 20 years conferred the greatest protection for both men and women (OR = 0.55, 95% CI: 0.39, 0.78 for men; OR = 0.44, 95% CI: 0.30, 0.67 for women). In summary, physical activity was associated with reduced risk of rectal cancer in these data. The reduced risk was similar to that previously observed for colon cancer. PMID- 12882944 TI - Relations of serum ascorbic acid and alpha-tocopherol to diabetic retinopathy in the Third National Health and Nutrition Examination Survey. AB - The protective relation of ascorbic acid and alpha-tocopherol to the development of diabetic retinopathy has not been thoroughly evaluated in epidemiologic studies. The association of prevalent diabetic retinopathy with serum ascorbic acid and alpha-tocopherol was studied among participants with type 2 diabetes (>or=40 years) (n = 998) in the Third National Health and Nutrition Examination Survey (1988-1994); 20% of the sample (n = 199) had prevalent retinopathy. The overall odds ratio for retinopathy among participants in quartile 4 compared with quartile 1 for serum ascorbic acid was 1.3 (95% confidence interval: 0.8, 2.3), with a p for trend = 0.60 after adjustment for the confounders of smoking, race, waist/hip ratio, hypertension, and duration of diabetes. The overall odds ratio for retinopathy among participants in quartile 4 compared with quartile 1 for serum alpha-tocopherol was 2.7 (95% confidence interval: 1.6, 4.6), with a p for trend = 0.14 after adjustment for confounders. After removal of supplement users of vitamin C (n = 307) or vitamin E (n = 298), the odds ratio changed direction or was attenuated: adjusted odds ratios for retinopathy among participants in quartile 4 compared with quartile 1 for serum ascorbic acid and alpha-tocopherol = 0.7 (95% confidence interval: 0.3, 1.4) and 1.6 (95% confidence interval: 0.9, 2.9), respectively. In summary, no significant associations were observed between serum levels of major dietary antioxidants and retinopathy. Recent use of supplements for treatment of complications of diabetes may explain the direct associations. PMID- 12882945 TI - Factors associated with increased and decreased risk of Campylobacter infection: a prospective case-control study in Norway. AB - In 1999-2000, a prospective case-control study of sporadic, domestically acquired campylobacteriosis was conducted in three counties in Norway to identify preventable risk factors and potentially protective factors. A total of 212 cases and 422 population controls matched by age, sex, and geographic area were enrolled. In conditional logistic regression analysis, the following factors were found to be independently associated with an increased risk of Campylobacter infection: drinking undisinfected water, eating at barbecues, eating poultry bought raw, having occupational exposure to animals, and eating undercooked pork. The following factors were independently related to a decreased risk: eating mutton, eating raw fruits or berries, and swimming. Results indicated that infection is more likely to occur as a result of cross-contamination from raw poultry products than because of poultry consumption per se. Drinking undisinfected water, reported by 53% of cases, was a leading risk factor in this study. Drinking water may constitute the common reservoir linking infection in humans and animals, including poultry and wild birds. Insight into the ecology of Campylobacter in freshwater ecosystems may be required to understand the epidemiology of campylobacteriosis. The possibility that certain foods confer protection against campylobacteriosis deserves exploration. PMID- 12882946 TI - Whole grain intake is associated with lower body mass and greater insulin sensitivity among adolescents. AB - The authors tested the hypothesis that consumption of whole grain is associated with greater insulin sensitivity and lower body mass index (BMI) (weight (kg)/height (m)(2)) in adolescents and that this association is stronger among the heaviest adolescents. Two 127-item food frequency questionnaires were administered at the mean ages of 13 years (standard deviation 1.2) and 15 years (standard deviation 1.3) to 285 Minnesota adolescents who underwent two euglycemic insulin clamp studies 2 years apart as part of a protocol evaluating the influence of insulin resistance on development of adverse cardiovascular disease risk factors. Intake of whole grain was examined for associations with BMI and insulin sensitivity (measured as milligrams of glucose uptake per kilogram of lean body mass (M(lbm)) per minute). After adjustment for age, gender, race, Tanner stage, and energy intake, mean BMI was 23.6 for adolescents consuming less than serving/day of whole-grain foods, 22.6 for -1 servings/day, and 21.9 for more than 1 servings/day (p = 0.05). After adjustment for age, gender, race, Tanner stage, energy intake, BMI, and physical activity, M(lbm) was 11.6, 12.3, and 13.2 mg/kg/minute, respectively, in the three whole grain intake groups (p = 0.02). This relation was stronger among adolescents with higher BMIs (p = 0.001). Whole grain intake was associated with greater insulin sensitivity and lower BMI in adolescents, especially among the heaviest persons. PMID- 12882947 TI - Interaction as departure from additivity in case-control studies: a cautionary note. AB - It has been argued that assessment of interaction should be based on departures from additive rates or risks. The corresponding fundamental interaction parameter cannot generally be estimated from case-control studies. Thus, surrogate measures of interaction based on relative risks from logistic models have been proposed, such as the relative excess risk due to interaction (RERI), the attributable proportion due to interaction (AP), and the synergy index (S). In practice, it is usually necessary to include covariates such as age and gender to control for confounding. The author uncovers two problems associated with surrogate interaction measures in this case: First, RERI and AP vary across strata defined by the covariates, whereas the fundamental interaction parameter is unvarying. S does not vary across strata, which suggests that it is the measure of choice. Second, a misspecification problem implies that measures based on logistic regression only approximate the true measures. This problem can be rectified by using a linear odds model, which also enables investigators to test whether the fundamental interaction parameter is zero. A simulation study reveals that coverage is much improved by using the linear odds model, but bias may be a concern regardless of whether logistic regression or the linear odds model is used. PMID- 12882948 TI - Selection bias in the assessment of gene-environment interaction in case-control studies. AB - Selection bias is a common concern in epidemiologic studies, particularly case control studies. Selection bias in the odds ratio occurs when participation depends jointly on exposure and disease status. General results for understanding when selection bias may affect studies involving gene-environment interactions have not yet been developed. In this paper, the authors show that the assessment of gene-environment interactions will not be subject to selection bias under the assumption that genotype does not influence participation conditional on exposure and disease status. This is true even when selection, including self-selection of subjects, is jointly influenced by exposure and disease and regardless of whether the genotype is related to exposure, disease, or both. The authors present an example to illustrate this concept. PMID- 12882949 TI - Validation of self-reported screening mammography histories among women with and without breast cancer. AB - As part of a case-control study of the efficacy of screening mammography, the authors validated the mammography histories of 2,495 women aged 40-64 years with incident breast cancer diagnosed in 1994-1998 and a 25% random sample of 615 controls never diagnosed with breast cancer, all reporting a mammogram in the past 5 years. Subjects from five metropolitan areas of the United States were cross-classified by facility records ("gold standard") and self-report according to history of a recent screening mammogram (within 1 year or within 2 years). Sensitivity and specificity of self-reported screening at 1 year were 0.93 and 0.82, respectively, for cases and 0.92 and 0.80 for controls. At 2 years, sensitivity and specificity were 0.97 and 0.78 for both cases and controls. Confidence intervals for the differences in sensitivity and specificity were narrow and included zero. Scant evidence was found of telescoping (recollection of events as more recent than actual). Findings suggest that, in an interview based case-control study of the efficacy of screening mammography, 1) estimated true prevalences of recent screening mammography adjusted for sensitivity and specificity will be slightly lower than self-reported prevalences, and 2) differential misclassification of exposure status is slight. Therefore, odds ratios will likely be biased toward the null, underestimating screening efficacy. PMID- 12882950 TI - Population- and community-based recruitment of African Americans and Latinos: the San Francisco Bay Area Lung Cancer Study. AB - Empiric data on recruitment of minorities into clinical or population studies are limited. The authors evaluated population- and community-based recruitment methods in a 1998-2001 case-control study of lung cancer among African Americans and Latinos. For lung cancer cases in the San Francisco Bay Area of California, rapid case ascertainment by the tumor registry combined with telephone screening identified 470 (9%) African Americans and 262 (5%) Latinos. When random digit dialing (RDD) and Health Care Financing Administration (HCFA) records failed to yield adequate numbers of controls in appropriate age-gender-ethnicity groups, community-based recruitment methods were used. Demographic characteristics and behavioral and occupational risk factors for controls, by recruitment method, were compared with those for lung cancer cases to evaluate potential bias. The average numbers of hours spent per control recruited were 18.6 for RDD, 11.4 for HCFA, and less than 1 for the community-based methods. The prevalence of smoking related lung cancer risk factors was significantly higher among African-American community-based controls than for those identified through RDD (p < 0.005). Compared with HCFA controls, Latino RDD controls reported significantly higher cumulative smoking exposure (p < 0.05). Further assessment of strategies for successful recruitment of minority participants into epidemiologic studies is warranted. PMID- 12882951 TI - Comparison of logistic regression versus propensity score when the number of events is low and there are multiple confounders. AB - The aim of this study was to use Monte Carlo simulations to compare logistic regression with propensity scores in terms of bias, precision, empirical coverage probability, empirical power, and robustness when the number of events is low relative to the number of confounders. The authors simulated a cohort study and performed 252,480 trials. In the logistic regression, the bias decreased as the number of events per confounder increased. In the propensity score, the bias decreased as the strength of the association of the exposure with the outcome increased. Propensity scores produced estimates that were less biased, more robust, and more precise than the logistic regression estimates when there were seven or fewer events per confounder. The logistic regression empirical coverage probability increased as the number of events per confounder increased. The propensity score empirical coverage probability decreased after eight or more events per confounder. Overall, the propensity score exhibited more empirical power than logistic regression. Propensity scores are a good alternative to control for imbalances when there are seven or fewer events per confounder; however, empirical power could range from 35% to 60%. Logistic regression is the technique of choice when there are at least eight events per confounder. PMID- 12882952 TI - Re: "Differential association of body mass index and fat distribution with three major histologic types of lung cancer: evidence from a cohort of older women". PMID- 12882953 TI - Re: "Statistical analysis of correlated data using generalized estimating equations: an orientation". PMID- 12882954 TI - Re: "Statistical analysis of correlated data using generalized estimating equations: an orientation". PMID- 12882957 TI - Role of carboxylate residues adjacent to the conserved core Walker B motifs in the catalytic cycle of multidrug resistance protein 1 (ABCC1). AB - MRP1 belongs to subfamily "C" of the ABC transporter superfamily. The nucleotide binding domains (NBDs) of the C family members are relatively divergent compared with many ABC proteins. They also differ in their ability to bind and hydrolyze ATP. In MRP1, NBD1 binds ATP with high affinity, whereas NBD2 is hydrolytically more active. Furthermore, ATP binding and/or hydrolysis by NBD2 of MRP1, but not NBD1, is required for MRP1 to shift from a high to low affinity substrate binding state. Little is known of the structural basis for these functional differences. One minor structural difference between NBDs is the presence of Asp COOH-terminal to the conserved core Walker B motif in NBD1, rather than the more commonly found Glu present in NBD2. We show that the presence of Asp or Glu following the Walker B motif profoundly affects the ability of the NBDs to bind, hydrolyze, and release nucleotide. An Asp to Glu mutation in NBD1 enhances its hydrolytic capacity and affinity for ADP but markedly decreases transport activity. In contrast, mutations that eliminate the negative charge of the Asp side chain have little effect. The decrease in transport caused by the Asp to Glu mutation in NBD1 is associated with an inability of MRP1 to shift from high to low affinity substrate binding states. In contrast, mutation of Glu to Asp markedly increases the affinity of NBD2 for ATP while decreasing its ability to hydrolyze ATP and to release ADP. This mutation eliminates transport activity but potentiates the conversion from a high to low affinity binding state in the presence of nucleotide. These observations are discussed in the context of catalytic models proposed for MRP1 and other ABC drug transport proteins. PMID- 12882958 TI - Complexes of plasmid DNA with basic domain 47-57 of the HIV-1 Tat protein are transferred to mammalian cells by endocytosis-mediated pathways. AB - Arginine-rich peptides, penetratins, as part of a number of cellular and viral proteins, can penetrate across plasma membrane directly, without participation of endocytosis. We show that one of penetratins, the basic domain 47-57 of human immunodeficiency virus, type 1, transcription factor Tat (Tat peptide), is able to interact with plasmid DNA electrostatically. These interactions result in formation of polyelectrolytic complexes at various negative/positive charge ratios of plasmid DNA and Tat peptide. Plasmid DNA is capable of binding to Tat peptide up to 1.7-fold excess of the complex positive charge. The DNA-Tat complexes can be used for delivery of plasmid DNA into mammalian cells. Transfection efficacy of cultured cells by DNA-Tat complexes is stimulated by free Tat peptide, most likely because it protects DNA-Tat complexes from disruption by anionic proteoglycans of cellular surface. Our data strongly argue in favor of the endocytosis-dependent mechanism of DNA-Tat complex uptake by mammalian cells similarly to internalization of complexes of plasmid DNA with other polycationic carriers. Moreover, different cell lines use different endocytosis-mediated pathways for DNA-Tat complex internalization. Intravenous injections to mice of DNA-Tat complexes in comparison with injections of naked DNA showed an inhibitory effect of DNA-Tat complex positive charge on expression of transferred gene. A low level of foreign gene expression in the liver of mice injected intravenously with positively charged DNA-Tat complexes is accounted for by inactivation of DNA-Tat complexes in the bloodstream due to their interactions with serum albumin. These data should be taken into account in an attempt to develop versatile gene delivery systems based on penetratin application for human disease therapy. PMID- 12882959 TI - The apical loop of the HIV-1 TAR RNA hairpin is stabilized by a cross-loop base pair. AB - The TAR hairpin of the HIV-1 RNA genome is indispensable for trans-activation of the viral promoter and virus replication. The TAR structure has been studied extensively, but most attention has been directed at the three-nucleotide bulge that constitutes the binding site of the viral Tat protein. In contrast, the conformational properties of the apical loop have remained elusive. We performed biochemical studies and molecular dynamics simulations, which indicate that the TAR loop is structured and stabilized by a cross-loop base pair between residues C30 and G34. Mutational disruption of the cross-loop base pair results in reduced Tat response of the LTR promoter, which can be rescued by compensatory mutations that restore the base pair. Thus, Tat-mediated transcriptional activation depends on the structure of the TAR apical loop. The C30-G34 cross-loop base pair classes TAR in a growing family of hairpins with a structured loop that was recently identified in ribosomal RNA, tRNA, and several viral and cellular mRNAs. PMID- 12882960 TI - The inositol 5'-phosphatase SHIP-1 and the Src kinase Lyn negatively regulate macrophage colony-stimulating factor-induced Akt activity. AB - Upon encountering macrophage colony-stimulating factor (M-CSF), human monocytes undergo a series of cellular signaling events leading to an increase in Akt activity. However, the regulation of these events is not completely understood. Because the inositol 5'-phosphatase SHIP-1 is an important regulator of intracellular levels of phosphatidylinositol 3,4,5-trisphosphate, an important second messenger necessary for Akt activation, we hypothesized that SHIP-1 was involved in the regulation of M-CSF receptor (M-CSF-R)-induced Akt activation. In the human monocytic cell line, THP-1, SHIP-1 became tyrosine-phosphorylated following M-CSF activation in a Src family kinase-dependent manner. Transfection of 3T3-Fms cells, which express the human M-CSF-R, with wild-type SHIP-1 showed that SHIP-1 was necessary for the negative regulation of M-CSF-induced Akt activation. In THP-1 cells, SHIP-1 bound Lyn, independent of the kinase activity of Lyn, following M-CSF activation. Utilizing a glutathione S-transferase fusion protein, we found that SHIP-1 bound to Lyn via the SHIP-1 Src homology 2 domain. Furthermore, transfection of THP-1 cells with a wild-type SHIP-1 construct reduced NF-kappaB-dependent transcriptional activation of a reporter gene, whereas a SHIP-1 Src homology 2 domain construct resulted in an increase in NF kappaB activation. Additionally, in 3T3-Fms cells, Lyn enhanced the ability of SHIP-1 to regulate Akt activation by stabilizing SHIP-1 at the cellular membrane. Finally, macrophages isolated from both SHIP-1- and Lyn-deficient mice exhibited enhanced Akt phosphorylation following M-CSF stimulation. These data provide the first evidence of the involvement of both SHIP-1 and Lyn in the negative regulation of M-CSF-R-induced Akt activation. PMID- 12882961 TI - A novel gamma-hydroxybutyrate dehydrogenase: identification and expression of an Arabidopsis cDNA and potential role under oxygen deficiency. AB - In plants, gamma-aminobutyrate (GABA), a non-protein amino acid, accumulates rapidly in response to a variety of abiotic stresses such as oxygen deficiency. Under normoxia, GABA is catabolized to succinic semialdehyde and then to succinate with the latter reaction being catalyzed by succinic semialdehyde dehydrogenase (SSADH). Complementation of an SSADH-deficient yeast mutant with an Arabidopsis cDNA library enabled the identification of a novel cDNA (designated as AtGH-BDH for Arabidopsis thaliana gamma-hydroxybutyrate dehydrogenase), which encodes a 289-amino acid polypeptide containing an NADP-binding domain. Constitutive expression of AtGHBDH in the mutant yeast enabled growth on 20 mm GABA and significantly enhanced the cellular concentrations of gamma hydroxybutyrate, the product of the GHDBH reaction. These data confirm that the cDNA encodes a polypeptide with GHBDH activity. Arabidopsis plants subjected to flooding-induced oxygen deficiency for up to 4 h possessed elevated concentrations of gamma-hydroxybutyrate as well as GABA and alanine. RNA expression analysis revealed that GHBDH transcription was not up-regulated by oxygen deficiency. These findings suggest that GHBDH activity is regulated by the supply of succinic semialdehyde or by redox balance. It is proposed that GHBDH and SSADH activities in plants are regulated in a complementary fashion and that GHBDH and gamma-hydroxybutyrate function in oxidative stress tolerance. PMID- 12882962 TI - Reaction intermediate structures of 1-aminocyclopropane-1-carboxylate deaminase: insight into PLP-dependent cyclopropane ring-opening reaction. AB - The pyridoxal 5'-phosphate-dependent enzymes have been evolved to catalyze diverse substrates and to cause the reaction to vary. 1-Aminocyclopropane-1 carboxylate deaminase catalyzes the cyclopropane ring-opening reaction followed by deamination specifically. Since it was discovered in 1978, the enzyme has been widely investigated from the mechanistic and physiological viewpoints because the substrate is a precursor of the plant hormone ethylene and the enzymatic reaction includes a cyclopropane ring-opening. We have previously reported the crystal structure of the native enzyme. Here we report the crystal structures of the two reaction intermediates created by the mutagenesis complexed with the substrate. The substrate was validated in the active site of two forms: 1). covalent-bonded external aldimine with the coenzyme in the K51T form and 2). the non-covalent interaction around the coenzyme in the Y295F form. The orientations of the substrate in both structures were quite different form each other. In concert with other site-specific mutation experiments, this experiment revealed the ingenious and unique strategies that are used to achieve the specific activity. The substrate incorporated into the active site is reactivated by a two-phenol charge relay system to lead to the formation of a Schiff base with the coenzyme. The catalytic Lys51 residue may play a novel role to abstract the methylene proton from the substrate in cooperation with other factors, the carboxylate group of the substrate and the electron-adjusting apparatuses of the coenzyme. PMID- 12882963 TI - p38 Mitogen-activated protein kinase stabilizes mRNAs that contain cyclooxygenase 2 and tumor necrosis factor AU-rich elements by inhibiting deadenylation. AB - AU-rich elements (AREs) in 3'-untranslated regions of mRNAs confer instability. They target mRNAs for rapid deadenylation and degradation and may enhance decapping. The p38 MAPK pathway stabilizes many otherwise unstable ARE-containing mRNAs encoding proteins involved in inflammation; however, the mRNA decay step(s) regulated by the signaling pathway are unknown. To investigate whether it regulates deadenylation or the decay of the mRNA body, we used a tetracycline regulated beta-globin mRNA reporter system to transcribe pulses of mRNA of uniform length. We measured on Northern gels the migration of reporter mRNAs isolated from cells transfected only with reporter plasmid or co-transfected with an active mutant of MAPK kinase-6, and treated either with or without the p38 MAPK inhibitor SB 203580. Differences in migration were shown by RNase H mapping with oligo(dT) to be due to poly(A) shortening. Insertion of an ARE into the beta globin reporter mRNA promoted rapid deadenylation and decay of hypo-adenylated reporter mRNA. p38 MAPK activation inhibited the deadenylation of reporter mRNAs containing either the cyclooxygenase-2 or tumor necrosis factor AREs. The regulation of deadenylation by p38 MAPK was found to be specific because deadenylation of the beta-globin reporter mRNA either lacking an ARE or containing the c-Myc 3'-untranslated region (which is not p38 MAPK-responsive) was unaffected by p38 MAPK. It was concluded that the p38 MAPK pathway predominantly regulates deadenylation, rather than decay of the mRNA body, and this provides an explanation for why p38 MAPK regulates mRNA stability in some situations and translation in others. PMID- 12882964 TI - Phosphatidylinositol 3-kinase interacts with the adaptor protein Dab1 in response to Reelin signaling and is required for normal cortical lamination. AB - Reelin is a large secreted signaling protein that binds to two members of the low density lipoprotein receptor family, the apolipoprotein E receptor 2 and the very low density lipoprotein receptor, and regulates neuronal positioning during brain development. Reelin signaling requires activation of Src family kinases as well as tyrosine phosphorylation of the intracellular adaptor protein Disabled-1 (Dab1). This results in activation of phosphatidylinositol 3-kinase (PI3K), the serine/threonine kinase Akt, and the inhibition of glycogen synthase kinase 3beta, a protein that is implicated in the regulation of axonal transport. Here we demonstrate that PI3K activation by Reelin requires Src family kinase activity and depends on the Reelin-triggered interaction of Dab1 with the PI3K regulatory subunit p85alpha. Because the Dab1 phosphotyrosine binding domain can interact simultaneously with membrane lipids and with the intracellular domains of apolipoprotein E receptor 2 and very low density lipoprotein receptor, Dab1 is preferentially recruited to the neuronal plasma membrane, where it is phosphorylated. Efficient Dab1 phosphorylation and activation of the Reelin signaling cascade is impaired by cholesterol depletion of the plasma membrane. Using a neuronal migration assay, we also show that PI3K signaling is required for the formation of a normal cortical plate, a step that is dependent upon Reelin signaling. PMID- 12882965 TI - Base excision repair intermediates induce p53-independent cytotoxic and genotoxic responses. AB - DNA alkylation damage is primarily repaired by the base excision repair (BER) machinery in mammalian cells. In repair of the N-alkylated purine base lesion, for example, alkyl adenine DNA glycosylase (Aag) recognizes and removes the base, and DNA polymerase beta (beta-pol) contributes the gap tailoring and DNA synthesis steps. It is the loss of beta-pol-mediated 5'-deoxyribose phosphate removal that renders mouse fibroblasts alkylation-hypersensitive. Here we report that the hypersensitivity of beta-pol-deficient cells after methyl methanesulfonate-induced alkylation damage is wholly dependent upon glycosylase mediated initiation of repair, indicating that alkylated base lesions themselves are tolerated in these cells and demonstrate that beta-pol protects against accumulation of toxic BER intermediates. Further, we find that these intermediates are initially tolerated in vivo by a second repair pathway, homologous recombination, inducing an increase in sister chromatid exchange events. If left unresolved, these BER intermediates trigger a rapid block in DNA synthesis and cytotoxicity. Surprisingly, both the cytotoxic and genotoxic signals are independent of both the p53 response and mismatch DNA repair pathways, demonstrating that p53 is not required for a functional BER pathway, that the observed damage response is not part of the p53 response network, and that the BER intermediate-induced cytotoxic and genotoxic effects are distinct from the mechanism engaged in response to mismatch repair signaling. These studies demonstrate that, although base damage is repaired by the BER pathway, incomplete BER intermediates are shuttled into the homologous recombination pathway, suggesting possible coordination between BER and the recombination machinery. PMID- 12882966 TI - Crystal structure of Pasteurella haemolytica ferric ion-binding protein A reveals a novel class of bacterial iron-binding proteins. AB - Pasteurellosis caused by the Gram-negative pathogen Pasteurella haemolytica is a serious disease leading to death in cattle. To scavenge growth-limiting iron from the host, the pathogen utilizes the periplasmic ferric ion-binding protein A (PhFbpA) as a component of an ATP-binding cassette transport pathway. We report the 1.2-A structure of the iron-free (apo) form of PhFbpA, which is a member of the transferrin structural superfamily. The protein structure adopts a closed conformation, allowing us to reliably assign putative iron-coordinating residues. Based on our analysis, PhFbpA utilizes a unique constellation of binding site residues and anions to octahedrally coordinate an iron atom. A surprising finding in the structure is the presence of two formate anions on opposite sides of the iron-binding pocket. The formate ions tether the N- and C-terminal domains of the protein and stabilize the closed structure, also providing clues as to probable candidates for synergistic anions in the iron-loaded state. PhFbpA represents a new class of bacterial iron-binding proteins. PMID- 12882967 TI - The MinD membrane targeting sequence is a transplantable lipid-binding helix. AB - MinD is a ubiquitous ATPase that plays a crucial role in selection of the division site in eubacteria, chloroplasts, and probably also Archaea. It was recently demonstrated that membrane localization of MinD is mediated by an 8-12 residue C-terminal motif termed the membrane targeting sequence or MTS. In this study we show that the MinD MTS is a transplantable lipid-binding motif that can effectively target heterologous proteins to the cell membrane. We demonstrate that eubacterial MTSs interact directly with lipid bilayers as an amphipathic helix, with a distinct preference for anionic phospholipids. Moreover, we provide evidence that the phospholipid preference of each MTS, as well as its affinity for biological membranes, has been evolutionarily "tuned" to its specific role in different bacteria. We propose a model to describe how the MTS is coupled to ATP binding to regulate the reversible membrane association of Escherichia coli MinD during its pole-to-pole oscillation cycle. PMID- 12882968 TI - Unique error signature of the four-subunit yeast DNA polymerase epsilon. AB - We have purified wild type and exonuclease-deficient four-subunit DNA polymerase epsilon (Pol epsilon) complex from Saccharomyces cerevisiae and analyzed the fidelity of DNA synthesis by the two enzymes. Wild type Pol epsilon synthesizes DNA accurately, generating single-base substitutions and deletions at average error rates of 5' exonuclease activity is less accurate to a degree suggesting that wild type Pol epsilon proofreads at least 92% of base substitution errors and at least 99% of frameshift errors made by the polymerase. Surprisingly the base substitution fidelity of exonuclease-deficient Pol epsilon is severalfold lower than that of proofreading-deficient forms of other replicative polymerases. Moreover the spectrum of errors shows a feature not seen with other A, B, C, or X family polymerases: a high proportion of transversions resulting from T.dTTP, T.dCTP, and C.dTTP mispairs. This unique error specificity and amino acid sequence alignments suggest that the structure of the polymerase active site of Pol epsilon differs from those of other B family members. We observed both similarities and differences between the spectrum of substitutions generated by proofreading-deficient Pol epsilon in vitro and substitutions occurring in vivo in a yeast strain defective in Pol epsilon proofreading and DNA mismatch repair. We discuss the implications of these findings for the role of Pol epsilon polymerase activity in DNA replication. PMID- 12882969 TI - Arginine metabolism in the deep sea tube worm Riftia pachyptila and its bacterial endosymbiont. AB - The present study describes the distribution and properties of enzymes involved in arginine metabolism in Riftia pachyptila, a tubeworm living around deep sea hydrothermal vents and known to be engaged in a highly specific symbiotic association with a bacterium. The results obtained show that the arginine biosynthetic enzymes, carbamyl phosphate synthetase, ornithine transcarbamylase, and argininosuccinate synthetase are present in all of the tissues of the worm and in the bacteria. Thus, Riftia and its bacterial endosymbiont can assimilate nitrogen and carbon via this arginine biosynthetic pathway. The kinetic properties of ornithine transcarbamylase strongly suggest that neither Riftia nor the bacteria possess the catabolic form of this enzyme belonging to the arginine deiminase pathway, the absence of this pathway being confirmed by the lack of arginine deiminase activity. Arginine decarboxylase and ornithine decarboxylase are involved in the biosynthesis of polyamines such as putrescine and agmatine. These activities are present in the trophosome, the symbiont-harboring tissue, and are higher in the isolated bacteria than in the trophosome, indicating that these enzymes are of bacterial origin. This finding indicates that Riftia is dependent on its bacterial endosymbiont for the biosynthesis of polyamines that are important for its metabolism and physiology. These results emphasize a particular organization of the arginine metabolism and the exchanges of metabolites between the two partners of this symbiosis. PMID- 12882970 TI - The receptor tyrosine kinase regulator Sprouty1 is a target of the tumor suppressor WT1 and important for kidney development. AB - WT1 encodes a transcription factor involved in kidney development and tumorigenesis. Using representational difference analysis, we identified a new set of WT1 targets, including a homologue of the Drosophila receptor tyrosine kinase regulator, sprouty. Sprouty1 was up-regulated in cell lines expressing wild-type but not mutant WT1. WT1 bound to the endogenous sprouty1 promoter in vivo and directly regulated sprouty1 through an early growth response gene-1 binding site. Expression of Sprouty1 and WT1 overlapped in the developing metanephric mesenchyme, and Sprouty1, like WT1, plays a key role in the early steps of glomerulus formation. Disruption of Sprouty1 expression in embryonic kidney explants by antisense oligonucleotides reduced condensation of the metanephric mesenchyme, leading to a decreased number of glomeruli. In addition, sprouty1 was expressed in the ureteric tree and antisense-treated ureteric trees had cystic lumens. Therefore, sprouty1 represents a physiologically relevant target gene of WT1 during kidney development. PMID- 12882971 TI - A novel mitochondrial carnitine-acylcarnitine translocase induced by partial hepatectomy and fasting. AB - The carnitine-dependent transport of long-chain fatty acids is essential for fatty acid catabolism. In this system, the fatty acid moiety of acyl-CoA is transferred enzymatically to carnitine, and the resultant product, acylcarnitine, is imported into the mitochondrial matrix through a transporter named carnitine acylcarnitine translocase (CACT). Here we report a novel mammalian protein homologous to CACT. The protein, designated as CACL (CACT-like), is localized to the mitochondria and has palmitoylcarnitine transporting activity. The tissue distribution of CACL is similar to that of CACT; both are expressed at a higher level in tissues using fatty acids as fuels, except in the brain, where only CACL is expressed. In addition, CACL is induced by partial hepatectomy or fasting. Thus, CACL may play an important role cooperatively with its homologue CACT in a stress-induced change of lipid metabolism, and may be specialized for the metabolism of a distinct class of fatty acids involved in brain function. PMID- 12882972 TI - NFkappaB regulates plasma apolipoprotein A-I and high density lipoprotein cholesterol through inhibition of peroxisome proliferator-activated receptor alpha. AB - The levels of plasma HDL cholesterol and apoA-I in NFkappaB p50 subunit-deficient mice were significantly higher than those in wild-type mice under regular and high fat diets, without any significant difference in the level of total cholesterol. To examine the role of NFkappaBin lipid metabolism, we studied its effect on the regulation of apoA-I secretion from human hepatoma HepG2 cells. Lipopolysaccharide-induced activation of NFkappaB reduced the expression of apoA I mRNA and protein, whereas adenovirus-mediated expression of IkappaBalpha super repressor ameliorated the reduction. This IkappaBalpha-induced apoA-I increase was blocked by preincubation with MK886, a selective inhibitor of peroxisome proliferator-activated receptor alpha (PPARalpha), suggesting that NFkappaB inactivation induces apoA-I through activation of PPARalpha. To further support this idea, the expression of IkappaBalpha increased apoA-I promoter activity, and this increase was blocked by preincubation with MK886. Mutations in the putative PPARalpha-binding site in the apoA-I promoter or lack of the site abrogated these changes. Taking these results together, inhibition of NFkappaB increases apoA-I and HDL cholesterol through activation of PPARalpha in vivo and in vitro. Our data suggest a new aspect of lipid metabolism and may lead to a new paradigm for prevention and treatment of atherosclerotic disease. PMID- 12882973 TI - Repair of UV lesions in silenced chromatin provides in vivo evidence for a compact chromatin structure. AB - Genes positioned close to telomeres in yeast are silenced by a heterochromatin like structure containing Sir proteins. To investigate whether silencing also affects DNA repair, we studied removal of UV lesions by photolyase and nucleotide excision repair (NER) in strains containing the URA3 gene inserted 2 kilobases from a telomere. URA3 was transcriptionally active in sir3delta mutants, partially silenced in SIR3 cells, or completely silenced by overexpression of SIR3 or deletion of RPD3. The active URA3 showed efficient repair by both pathways. Fast repair of the promoter and 3' end by photolyase reflected a non nucleosomal structure. Partial silencing had no remarkable effect on photolyase but reduced repair by NER, indicating differential accessibility for the two repair reactions. Complete silencing inhibits NER and photolyase in the coding region as well as in the promoter and the 3'-end. Conventional nuclease footprinting analyses revealed subtle changes in the promoter proximal nucleosome under partially silenced conditions but a pronounced reorganization of chromatin extending over the whole gene in silenced chromatin. Thus, both repair systems are sensitive to chromatin changes associated with silencing and provide direct evidence for a compact structure of heterochromatin. PMID- 12882974 TI - Cloning, expression, characterization, and interaction of two components of a human mitochondrial fatty acid synthase. Malonyltransferase and acyl carrier protein. AB - The possibility that human cells contain, in addition to the cytosolic type I fatty acid synthase complex, a mitochondrial type II malonyl-CoA-dependent system for the biosynthesis of fatty acids has been examined by cloning, expressing, and characterizing two putative components. Candidate coding sequences for a malonyl CoA:acyl carrier protein transacylase (malonyltransferase) and its acyl carrier protein substrate, identified by BLAST searches of the human sequence data base, were located on nuclear chromosomes 22 and 16, respectively. The encoded proteins localized exclusively in mitochondria only when the putative N-terminal mitochondrial targeting sequences were present as revealed by confocal microscopy of HeLa cells infected with appropriate green fluorescent protein fusion constructs. The mature, processed forms of the mitochondrial proteins were expressed in Sf9 cells and purified, the acyl carrier protein was converted to the holoform in vitro using purified human phosphopantetheinyltransferase, and the functional interaction of the two proteins was studied. Compared with the dual specificity malonyl/acetyltransferase component of the cytosolic type I fatty acid synthase, the type II mitochondrial counterpart exhibits a relatively narrow substrate specificity for both the acyl donor and acyl carrier protein acceptor. Thus, it forms a covalent acyl-enzyme complex only when incubated with malonyl-CoA and transfers exclusively malonyl moieties to the mitochondrial holoacyl carrier protein. The type II acyl carrier protein from Bacillus subtilis, but not the acyl carrier protein derived from the human cytosolic type I fatty acid synthase, can also function as an acceptor for the mitochondrial transferase. These data provide compelling evidence that human mitochondria contain a malonyl-CoA/acyl carrier protein-dependent fatty acid synthase system, distinct from the type I cytosolic fatty acid synthase, that resembles the type II system present in prokaryotes and plastids. The final products of this system, yet to be identified, may play an important role in mitochondrial function. PMID- 12882975 TI - Monitoring active site alterations upon mutation of yeast pyruvate kinase using 205Tl+ NMR. AB - The interaction of the monovalent cation with wild type (WT) yeast pyruvate kinase (YPK) and with the T298S, T298C, and T298A mutants was investigated by 205Tl+ NMR to monitor possible structural alterations at the active site by Thr 298 mutation. TlNO3 activates WT YPK with a kcat value similar to that obtained with KCl and an apparent Ka of 0.96 +/- 0.07 mm in the presence of Mn2+ and fructose 1,6-bisphosphate. With the three mutants, Tl+ is a better activator than is K+ based on kcat values. Tl+ activation and inhibition of YPK is affected by mutation of the active site Thr-298. The effect of Mn2+ on the 1/T value of 205Tl+1 in the presence of the WT and mutant YPK complexes was determined at 173 MHz (300 MHz, 1H) and 346 MHz (600 MHz, 1H). For each complex studied, 1/pT2p >> 1/pT1p and 1/pT1p is frequency-dependent suggesting fast exchange conditions. The values of 1/pT1p differ for each mutant. A correlation time of 0.65 +/- 0.35 ns was estimated for the Mn2+-205Tl+ interaction. The Tl+-Mn2+ distances at the active site of YPK were calculated from the paramagnetic contribution of Mn2+ to 1/T1M of YPK-bound 205Tl+. The calculated Tl+-Mn2+ distance for the Thr-298 mutants is decreased by about 1 A from 6.0 +/- 0.2 A observed with WT. The results suggest conformational alterations at the active site of YPK where phosphoryl transfer occurs upon mutation of Thr-298. These conformational changes may, in part, explain the alteration in kcat and kcat/Km,PEP observed with the Thr-298 mutants. PMID- 12882976 TI - Juxtaposition of the two distal CX3C motifs via intrachain disulfide bonding is essential for the folding of Tim10. AB - The TIM10 complex, composed of the homologous proteins Tim10 and Tim9, chaperones hydrophobic proteins inserted at the mitochondrial inner membrane. A salient feature of the TIM10 complex subunits is their conserved "twin CX3C" motif. Systematic mutational analysis of all cysteines of Tim10 showed that their underlying molecular defect is impaired folding (demonstrated by circular dichroism, aberrant homo-oligomer formation, and thiol trapping assays). As a result of defective folding, clear functional consequences were manifested in (i) complex formation with Tim9, (ii) chaperone activity, and (iii) import into tim9ts mitochondria lacking both endogenous Tim9 and Tim10. The organization of the four cysteines in intrachain disulfides was determined by trypsin digestion and mass spectrometry. The two distal CX3C motifs are juxtaposed in the folded structure and disulfide-bonded to each other rather than within each other, with an inner cysteine pair connecting Cys44 with Cys61 and an outer pair between Cys40 and Cys65. These cysteine pairs are not equally important for folding and assembly; mutations of the inner Cys are severely affected and form wrong, non native disulfides, in contrast to mutations of the outer Cys that can still maintain the native inner disulfide pair and display weaker functional defects. Taken together these data reveal this specific intramolecular disulfide bonding as the crucial mechanism for Tim10 folding and show that the inner cysteine pair has a more prominent role in this process. PMID- 12882977 TI - Platelet-derived growth factor-BB-mediated activation of Akt suppresses smooth muscle-specific gene expression through inhibition of mitogen-activated protein kinase and redistribution of serum response factor. AB - Platelet-derived growth factor (PDGF) inhibits expression of smooth muscle (SM) genes in vascular smooth muscle cells and blocks induction by arginine vasopressin (AVP). We have previously demonstrated that suppression of SM-alpha actin by PDGF-BB is mediated in part through a Ras-dependent pathway. This study examined the role of phosphatidylinositol 3-kinase (PI3K)y and its downstream effector, Akt, in regulating SM gene expression. PDGF caused a rapid sustained activation of Akt, whereas AVP caused only a small transient increase. PDGF selectively caused a sustained stimulation of p85/p110 alpha PI3K. In contrast, p85/110 beta PI3K activity was not altered by either PDGF or AVP, whereas both agents caused a delayed activation of Class IB p101/110 gamma PI3K. Expression of a gain-of-function PI3K or myristoylated Akt (myr-Akt) mimicked the inhibitory effect of PDGF on SM-alpha-actin and SM22 alpha expression. Pretreatment with LY 294002 reversed the inhibitory effect of PDGF. Expression of myr-Akt selectively inhibited AVP-induced activation of c-Jun N-terminal kinase and p38 mitogen activated protein kinases, which we have shown are critical for induction of these genes. Nuclear extracts from PDGF-stimulated or myr-Akt expressing cells showed reduced serum response factor binding to SM-specific CArG elements. This was associated with appearance of serum response factor in the cytoplasm. These data indicate that activation of p85/p110 alpha/Akt mediates suppression of SM gene expression by PDGF. PMID- 12882978 TI - Lysine 238 is an essential residue for alpha,beta-elimination catalyzed by Treponema denticola cystalysin. AB - Treponema denticola cystalysin is a pyridoxal 5'-phosphate (PLP) enzyme that catalyzes the alpha,beta-elimination of l-cysteine to pyruvate, ammonia, and H2S. Similar to other PLP enzymes, an active site Lys residue (Lys-238) forms an internal Schiff base with PLP. The mechanistic role of this residue has been studied by an analysis of the mutant enzymes in which Lys-238 has been replaced by Ala (K238A) and Arg (K238R). Both apomutants reconstituted with PLP bind noncovalently approximately 50% of the normal complement of the cofactor and have a lower affinity for the coenzyme than that of wild-type. Kinetic analyses of the reactions of K238A and K238R mutants with glycine compared with that of wild-type demonstrate the decrease of the rate of Schiff base formation by 103- and 7.5 x 104-fold, respectively, and, to a lesser extent, a decrease of the rate of Schiff base hydrolysis. Thus, a role of Lys-238 is to facilitate formation of external aldimine by transimination. Kinetic data reveal that the K238A mutant is inactive in the alpha,beta-elimination of l-cysteine and beta-chloro-l-alanine, whereas K238R retains 0.3% of the wild-type activity. These data, together with those derived from a spectral analysis of the reaction of Lys-238 mutants with unproductive substrate analogues, indicate that Lys-238 is an essential catalytic residue, possibly participating as a general base abstracting the Calpha-proton from the substrate and possibly as a general acid protonating the beta-leaving group. PMID- 12882979 TI - TL1A-induced NF-kappaB activation and c-IAP2 production prevent DR3-mediated apoptosis in TF-1 cells. AB - We recently identified TL1A, an endothelium-derived T cell costimulator and a ligand for tumor necrosis factor receptor superfamily members DR3 and decoy receptor 3. To elucidate the signaling events triggered by TL1A-DR3 interaction and to understand the molecular mechanisms regulating DR3-mediated apoptosis, we have studied the effect of TL1A and an agonistic DR3 monoclonal antibody in human erythroleukemic TF-1 cells, which express DR3 endogenously. TL1A induced the formation of a DR3 signaling complex containing TRADD, TRAF2, and RIP and activated the NF-kappaB and the ERK, JNK, and p38 mitogen-activated protein kinase pathways. However, TL1A or an agonistic DR3 monoclonal antibody did not induce apoptosis in these cells nor were there detectable levels of FADD or procaspase-8 seen in the signaling complex. Interestingly, DR3-mediated apoptosis was induced in TF-1 cells in the presence of a NF-kappaB pathway-specific inhibitor but not in the presence of mitogen-activated protein kinase inhibitors, either alone or in combination, suggesting that DR3-induced NF-kappaB activation was responsible for resistance to apoptosis in these cells. Consistent with this, we found that TL1A significantly increased the production of c-IAP2, a known NF kappaB-dependent anti-apoptotic protein, and that the NF-kappaB inhibitor or cycloheximide prevented its synthesis. Furthermore, inhibition of c-IAP2 production by RNA interference significantly sensitized TF-1 cells to TL1A induced apoptosis. Our study identifies a molecular mechanism by which TL1A and DR3 regulate cell fate in TF-1 cells. PMID- 12882980 TI - High affinity for farnesyltransferase and alternative prenylation contribute individually to K-Ras4B resistance to farnesyltransferase inhibitors. AB - Farnesyltransferase inhibitors (FTIs) block Ras farnesylation, subcellular localization and activity, and inhibit the growth of Ras-transformed cells. Although FTIs are ineffective against K-Ras4B, the Ras isoform most commonly mutated in human cancers, they can inhibit the growth of tumors containing oncogenic K-Ras4B, implicating other farnesylated proteins or suggesting distinct functions for farnesylated and for geranylgeranylated K-Ras, which is generated when farnesyltransferase is inhibited. In addition to bypassing FTI blockade through geranylgeranylation, K-Ras4B resistance to FTIs may also result from its higher affinity for farnesyltransferase. Using chimeric Ras proteins containing all combinations of Ras background, CAAX motif, and K-Ras polybasic domain, we show that either a polybasic domain or an alternatively prenylated CAAX renders Ras prenylation, Ras-induced Elk-1 activation, and anchorage-independent cell growth FTI-resistant. The polybasic domain alone increases the affinity of Ras for farnesyltransferase, implying independent roles for each K-Ras4B sequence element in FTI resistance. Using microarray analysis and colony formation assays, we confirm that K-Ras function is independent of the identity of the prenyl group and, therefore, that FTI inhibition of K-Ras transformed cells is likely to be independent of K-Ras inhibition. Our results imply that relevant FTI targets will lack both polybasic and potentially geranylgeranylated methionine-CAAX motifs. PMID- 12882981 TI - The unique hexokinase of Kluyveromyces lactis. Molecular and functional characterization and evaluation of a role in glucose signaling. AB - The Crabtree-negative yeast Kluyveromyces lactis is capable of adjusting its glycolytic flux to the requirements of respiration by tightly regulating glucose uptake. RAG5 encoding the only glucose and fructose phosphorylating enzyme present in K. lactis is required for the up-regulation of glucose transport and also for glucose repression. To understand the significance of the molecular identity and specific function(s) of the corresponding kinase to glucose signaling, RAG5 was overexpressed and its gene product KlHxk1 (Rag5p) isolated and characterized. Stopped-flow kinetics and sedimentation analysis indicated a monomer-homodimer equilibrium of KlHxk1 in a condition of catalysis, i.e. in the presence of substrates and products. The kinetic constants of ATP-dependent glucose phosphorylation identified a 53-kDa monomer as the high affinity/high activity form of the novel enzyme for both glycolytic substrates suggesting a control of glucose phosphorylation at the level of dimer formation and dissociation. In contrast to the highly homologous hexokinase isoenzyme 2 of Saccharomyces cerevisiae (ScHxk2), KlHxk1 was not inhibited by free ATP in a physiological range of nucleotide concentration. Mass spectrometric sequencing of tryptic peptides of KlHxk1 identified unmodified serine at amino acid position 156. The corresponding amino acid in ScHxk2 is serine 157, which represents the autophosphorylation-inactivation site. KlHxk1 did not display, however, the typical pattern of inactivation under the respective in vitro conditions and maintained a high residual glucose phosphorylating activity. The biophysical and functional data are discussed with respect to a possible regulatory role of KlHxk1 in glucose metabolism and signaling in K. lactis. PMID- 12882982 TI - A biochemical and molecular characterization of LEP1, an extensin peroxidase from lupin. AB - An analysis of apoplastic extensin cross-linking activity in vegetative organs of Lupinus albus indicated that leaves contained the highest specific activity. Assays of peroxidases fractionated from this material demonstrated that this activity could be largely attributed to a soluble and apoplastic 51-kDa peroxidase, denoted LEP1. Relative to other purified peroxidases, LEP1 demonstrates high extensin cross-linking activity and can be classified as an extensin peroxidase (EP). Optimal conditions for the in vitro oxidation of other phenolic substrates included 1.5-3.0 mm peroxide at pH 5.0. EP activity of LEP1 was low under these conditions but optimal and substantially higher with 100 microm peroxide and neutral pH, suggesting that physiological changes in pH and peroxide in muro could heavily influence the extensin cross-linking activity of LEP1 in vivo. Analysis of LEP1 glycans indicated 11-12 N-linked glycans, predominantly the heptasaccharide Man3XylFucGlcNAc2, but also larger structures showing substantial heterogeneity. Comparative assays with horseradish peroxidase isoform C and peanut peroxidases suggested the high level of glycosylation in LEP1 may be responsible for the high solubility of this EP in the apoplastic space. A full-length cDNA corresponding to LEP1 was cloned. Quantitative reverse transcriptase-PCR demonstrated LEP1 induction in apical portions of etiolated hypocotyls 30-60 min after exposure to white light, prior to the onset of growth inhibition. Comparative modeling of the translated sequence indicated an unusually unobstructed equatorial cleft across the substrate access channel, which might facilitate interaction with extensin and confer higher EP activity. PMID- 12882983 TI - Mapping of a conformational epitope shared between E1 and E2 on the serum-derived human hepatitis C virus envelope. AB - Monoclonal antibody D32.10 produced by immunizing mice with a hepatitis C virus (HCV)-enriched pellet obtained from plasmapheresis of a chronically HCV1b infected patient binds HCV particles derived from serum of different HCV1a- and HCV1b-infected patients. Moreover, this monoclonal has been shown to recognize both HCV envelope proteins E1 and E2. In an attempt to provide novel insight into the membrane topology of HCV envelope glycoproteins E1 and E2, we localized the epitope recognized by D32.10 on the E1 and/or E2 sequence using Ph.D.-12 phage display peptide library technology. Mimotopes selected from the phage display dodecapeptide library by D32.10 shared partial similarities with 297RHWTTQGCNC306 of the HCV E1 glycoprotein and with both 613YRLWHYPCT621 and 480PDQRPYCWHYPPKPC494 of the HCV E2 glycoprotein. Immunoreactivity of D32.10 with overlapping peptides corresponding to these three HCV regions confirmed these localizations and suggested that the three regions identified are likely closely juxtaposed on the surface of serum-derived particles as predicted by the secondary model structure of HCV E2 derived from the tick-borne encephalitis virus E protein. This assertion was supported by the detection of specific antibodies directed against these three E1E2 regions in sera from HCV-infected patients. PMID- 12882984 TI - Nucleophosmin interacts with and inhibits the catalytic function of eukaryotic initiation factor 2 kinase PKR. AB - In normal cells the protein kinase PKR effects apoptosis in response to various extra and intracellular cues and can also function to suppress the neoplastic phenotype. Because most neoplastic cells are resistant to certain apoptotic cues, we reasoned that an early molecular event in carcinogenesis or leukemogenesis might be the inactivation of PKR by expression or activation of intracellular PKR inhibitors. Seeking novel PKR-modulating proteins we report here that nucleophosmin (NPM), a protein frequently overexpressed in a variety of human malignancies, binds to PKR, and inhibits its activation. Co-immunoprecipitation and in vitro binding experiments showed that NPM associated with PKR. Kinase assays demonstrated that recombinant NPM inhibited PKR activation in a dose dependent manner. In addition, purified recombinant NPM was phosphorylated by activated PKR. Most importantly, overexpression of NPM suppressed PKR activity, enhanced protein synthesis, and inhibited apoptosis. Lymphoblasts from patients with Fanconi anemia (FA) expressed low levels of NPM, which correlated with high ground-state activation of PKR and cellular hypersensitivity to apoptotic cues, but enforced expression of NPM in these mutant cells reduced aberrant apoptotic responses. Inhibition of PKR by NPM may be one mechanism by which neoplastic clones evolve in sporadic malignancies and in neoplastic cells arising in the context of the cancer predisposition syndrome, Fanconi anemia. PMID- 12882985 TI - Essential requirement of apolipoprotein J (clusterin) signaling for IkappaB expression and regulation of NF-kappaB activity. AB - Apolipoprotein J/clusterin is an enigmatic protein highly regulated in inflammation, apoptosis, and cancer. Despite extensive studies, its biological function has remained obscure. Here we show that apolipoprotein J inhibits neuroblastoma cell invasion. Since this function can be regulated by NF-kappaB, we explored the possibility that apolipoprotein J might interfere with NF-kappaB signaling. Ectopic apolipoprotein J expression strongly inhibited NF-kappaB activity in human neuroblastoma cells and murine embryonic fibroblasts by stabilizing inhibitors of NF-kappaB (IkappaBs). Steady state levels of IkappaB proteins are drastically reduced in mouse embryo fibroblasts after disruption of the apolipoprotein J gene. Absence of apolipoprotein J causes reduction of IkappaB stability, a tumor necrosis factor-dependent increase in NF-kappaB activity, increased transcription of the NF-kappaB target gene c-IAP and down modulation of p53 protein. These results suggest that an unexpected physiological role of apolipoprotein J is to inhibit NF-kappaB signaling through stabilization of IkappaBs and that this activity may result in suppression of tumor cell motility. PMID- 12882986 TI - Energy cost of walking and gait instability in healthy 65- and 80-yr-olds. AB - This study tested whether the lower economy of walking in healthy elderly subjects is due to greater gait instability. We compared the energy cost of walking and gait instability (assessed by stride to stride changes in the stride time) in octogenarians (G80, n = 10), 65-yr-olds (G65, n = 10), and young controls (G25, n = 10) walking on a treadmill at six different speeds. The energy cost of walking was higher for G80 than for G25 across the different walking speeds (P < 0.05). Stride time variability at preferred walking speed was significantly greater in G80 (2.31 +/- 0.68%) and G65 (1.93 +/- 0.39%) compared with G25 (1.40 +/- 0.30%; P < 0.05). There was no significant correlation between gait instability and energy cost of walking at preferred walking speed. These findings demonstrated greater energy expenditure in healthy elderly subjects while walking and increased gait instability. However, no relationship was noted between these two variables. The increase in energy cost is probably multifactorial, and our results suggest that gait instability is probably not the main contributing factor in this population. We thus concluded that other mechanisms, such as the energy expenditure associated with walking movements and related to mechanical work, or neuromuscular factors, are more likely involved in the higher cost of walking in elderly people. PMID- 12882987 TI - Change in spontaneous baroreflex control of pulse interval during heat stress in humans. AB - Spontaneous baroreflex control of pulse interval (PI) was assessed in healthy volunteers under thermoneutral and heat stress conditions. Subjects rested in the supine position with their lower legs in a water bath at 34 degrees C. Heat stress was imposed by increasing the bath temperature to 44 degrees C. Arterial blood pressure (Finapres), PI (ECG), esophageal and skin temperature, and stroke volume were continuously collected during each 5-min experimental stage. Spontaneous baroreflex function was evaluated by multiple techniques, including 1) the mean slope of the linear relationship between PI and systolic blood pressure (SBP) with three or more simultaneous increasing or decreasing sequences, 2) the linear relationship between changes in PI and SBP (deltaPI/DeltaSBP) derived by using the first differential equation, 3) the linear relationship between changes in PI and SBP with simultaneously increasing or decreasing sequences (+deltaPI/+deltaSBP or -deltaPI/-deltaSBP), and 4) transfer function analysis. Heat stress increased esophageal temperature by 0.6 +/- 0.1 degrees C, decreased PI from 1,007 +/- 43 to 776 +/- 37 ms and stroke volume by 16 +/- 5 ml/beat. Heat stress reduced baroreflex sensitivity but increased the incidence of baroreflex slopes from 5.2 +/- 0.8 to 8.6 +/- 0.9 sequences per 100 heartbeats. Baroreflex sensitivity was significantly correlated with PI or vagal power (r2 = 0.45, r2 = 0.71, respectively; P < 0.05). However, the attenuation in baroreflex sensitivity during heat stress appeared related to a shift in autonomic balance (shift in resting PI) rather than heat stress per se. PMID- 12882988 TI - Octanoate oxidation measured by 13C-NMR spectroscopy in rat skeletal muscle, heart, and liver. AB - Contribution of octanoate to the oxidative metabolism of the major sites of fatty acid oxidation (heart, liver, and resting and contracting skeletal muscle) was assessed in the intact rat with 13C-NMR spectroscopy. Under inhalation anesthesia, [2,4,6,8-13C4]octanoate was infused into the jugular vein and the sciatic nerve of one limb was stimulated for 1 h. Octanoate was a principal contributor to the acetyl-CoA pool in all tissues examined, with highest oxidation occurring in heart and soleus muscle followed by predominantly red portion of gastrocnemius muscle (RG), liver, and then white portion of gastrocnemius muscle (WG). Fractional contribution of 13C-labeled octanoate to the acetyl-CoA pool (Fc2) was 0.563 +/- 0.066 for heart and 0.367 +/- 0.054 for liver. Significant differences were observed between each of the muscle types during both rest and contraction. In muscle, Fc2 was highest in soleus (0.565 +/- 0.089 rested, 0.564 +/- 0.096 contracted), followed by RG (0.470 +/- 0.092 rested, 0.438 +/- 0.072 contracted), and lowest in WG (0.340 +/- 0.081 rested, 0.272 +/- 0.065 contracted). Our findings demonstrate that the fractional contribution of octanoate to oxidative metabolism correlates with oxidative capacity of the tissue and that octanoate metabolism increases in contracted muscle in proportion to the overall increase in oxidative rate. PMID- 12882989 TI - Na+-K+-ATPase in rat skeletal muscle: content, isoform, and activity characteristics. AB - The purpose of this study was to investigate the hypothesis that muscle Na+-K+ ATPase activity is directly related to Na+-K+-ATPase content and the content of the alpha2-catalytic isoform in muscles of different fiber-type composition. To investigate this hypothesis, tissue was sampled from soleus (Sol), red gastrocnemius (RG), white gastrocnemius (WG), and extensor digitorum longus (EDL) muscles at rest from 38 male Wistar rats weighing 413 +/- 6.0 g (mean +/- SE). Na+-K+-ATPase activity was determined in homogenates (Hom) and isolated crude membranes (CM) by the regenerating ouabain-inhibitable hydrolytic activity assay (ATPase) and the 3-O-methylfluorescein K+-stimulated phosphatase (3-O-MFPase) assay in vitro. In addition, Na+-K+-ATPase content (Bmax) and the distribution of alpha1-, alpha2-, beta1-, and beta2-isoforms were determined by [3H]ouabain binding and Western blot, respectively. For the ATPase assay, differences (P < 0.05) in enzyme activity between muscles were observed in Hom (EDL > WG) and in CM (Sol > EDL = WG). For the 3-O-MFPase assay, differences (P < 0.05) were also found for Hom (Sol > RG = EDL > WG) and CM (Sol = WG > RG). For Bmax, differences in the order of RG = EDL > Sol = WG (P < 0.05) were observed. Isoform distribution was similar between Hom and CM and indicated in CM, a greater density (P < 0.05) of alpha1 in Sol than WG and EDL (P < 0.05), but more equal distribution of alpha2 between muscles. The beta1 was greater (P < 0.05) in Sol and RG, and the beta2 was greater in EDL and WG (P < 0.05). Over all muscles, the correlation (r) between Hom 3-O-MFPase and Bmax was 0.45 (P < 0.05) and between Hom alpha2 and Bmax, 0.59 (P < 0.05). The alpha1 distribution correlated to Hom 3 O-MFPase (r = 0.79, P < 0.05) CM ATPase (r = 0.69, P < 0.005) and CM 3-O-MFPase activity (r = 0.32, P < 0.05). The alpha2 distribution was not correlated with any of the Na+-K+-ATPase activity measurements. The results indicate generally poor relationships between activity and total pump content and alpha2 isoform content of the Na+-K+-ATPase. Several factors, including the type of preparation and the type of assay, appear important in this regard. PMID- 12882990 TI - Skeletal muscle adaptations to microgravity exposure in the mouse. AB - To investigate the effects of microgravity on murine skeletal muscle fiber size, muscle contractile protein, and enzymatic activity, female C57BL/6J mice, aged 64 days, were divided into animal enclosure module (AEM) ground control and spaceflight (SF) treatment groups. SF animals were flown on the space shuttle Endeavour (STS-108/UF-1) and subjected to approximately 11 days and 19 h of microgravity. Immunohistochemical analysis of muscle fiber cross-sectional area revealed that, in each of the muscles analyzed, mean muscle fiber cross-sectional area was significantly reduced (P < 0.0001) for all fiber types for SF vs. AEM control. In the soleus, immunohistochemical analysis of myosin heavy chain (MHC) isoform expression revealed a significant increase in the percentage of muscle fibers expressing MHC IIx and MHC IIb (P < 0.05). For the gastrocnemius and plantaris, no significant changes in MHC isoform expression were observed. For the muscles analyzed, no alterations in MHC I or MHC IIa protein expression were observed. Enzymatic analysis of the gastrocnemius revealed a significant decrease in citrate synthase activity in SF vs. AEM control. PMID- 12882991 TI - O2 uptake kinetics during exercise at peak O2 uptake. AB - Compared with moderate- and heavy-intensity exercise, the adjustment of O2 uptake (VO2) to exercise intensities that elicit peak VO2 has received relatively little attention. This study examined the VO2 response of 21 young, healthy subjects (25 +/- 6 yr; mean +/- SD) during cycle ergometer exercise to step transitions in work rate (WR) corresponding to 90, 100, and 110% of the peak WR achieved during a preliminary ramp protocol (15-30 W/min). Gas exchange was measured breath by breath and interpolated to 1-s values. VO2 kinetics were determined by use of a two- or three-component exponential model to isolate the time constant (tau2) as representative of VO2 kinetics and the amplitude (Amp) of the primary fast component independent of the appearance of any VO2 slow component. No difference in VO2 kinetics was observed between WRs (tau90 = 24.7 +/- 9.0; tau100 = 22.8 +/- 6.7; tau110 = 21.5 +/- 9.2 s, where subscripts denote percent of peak WR; P > 0.05); nor in a subgroup of eight subjects was tau2 different from the value for moderate-intensity ( 0.05). As expected, the Amp increased with increasing WRs (Amp90 = 2,089 +/- 548; Amp100 = 2,165 +/- 517; Amp110 = 2,225 +/- 559 ml/min; Amp90 vs. Amp110, P < 0.05). However, the gain (G) of the VO2 response (deltaVO2/deltaWR) decreased with increasing WRs (G90 = 8.5 +/- 0.6; G100 = 7.9 +/- 0.6; G110 = 7.3 +/- 0.6 ml.min 1.W-1; P < 0.05). The Amp of the primary component approximated 85, 88, and 89% of peak Vo2 during 90, 100, and 110% WR transitions, respectively. The results of the present study demonstrate that, compared with moderate- and heavy-intensity exercise, the gain of the Vo2 response (as deltaVO2/DeltaWR) is reduced for exercise transitions in the severe-intensity domain, but the approach to this gain is well described by a common time constant that is invariant across work intensities. The lower deltaVO2/deltaWR may be due to an insufficient adjustment of the cardiovascular and/or pulmonary systems that determine O2 delivery to the exercising muscles or due to recruitment of motor units with lower oxidative capacity, after the onset of exercise in the severe-intensity domain. PMID- 12882992 TI - Influence of age and run training on cardiac Na+/Ca2+ exchange. AB - Effects of age and training on myocardial Na+/Ca2+ exchange were examined in young sedentary (YS; 14-15 mo), aged sedentary (AS; 27-31 mo), and aged trained (AT; 8- to 11-wk treadmill run training) male Fischer Brown Norway rats. Whole heart performance and isolated cardiocyte Na+/Ca2+ exchange characteristics were measured. At the whole heart level, a small but significant slowing of late isovolumic left ventricular (LV) relaxation, which may be indicative of altered Na+/Ca2+ exchange activity, was seen in hearts from AS rats. This subtle impairment in relaxation was not observed in hearts from AT rats. At the single cardiocyte level, late action potential duration was prolonged, resting membrane potential was more positive, and overshoot potential was greater in cardiocytes from AS rats than from YS rats (P < 0.05). Training did not influence any of these age-related action potential characteristics. In electrically paced cardiocytes, neither shortening nor intracellular Ca2+ concentration ([Ca2+]i) dynamics was influenced by age or training. Similarly, neither age nor training influenced the rate of [Ca2+]i clearance via forward (Nain+ /Caout2+) Na+/Ca2+ exchange after caffeine-induced Ca2+ release from the sarcoplasmic reticulum or cardiac Na+/Ca2+ exchanger protein (NCX1) expression. However, when whole cell patch-clamp techniques combined with fluorescence microscopy were used to evaluate the ability of Na+/Ca2+ exchange to alter cytosolic [Ca2+] ([Ca2+]c) under conditions where membrane potential (Vm) and internal and external [Na+] and [Ca2+] could be controlled, we observed age-associated increases in forward Na+/Ca2+ exchange-mediated [Ca2+]c clearance (P < 0.05) that were not influenced by training. The age-related increase in forward Na+/Ca2+ exchange activity provides a hypothetical explanation for the late action potential prolongation observed in this study. PMID- 12882993 TI - Impaired leg vasodilation during dynamic exercise in healthy older women. AB - The purpose of the present study was to test the hypothesis that leg blood flow responses during leg cycle ergometry are reduced with age in healthy non-estrogen replaced women. Thirteen younger (20-27 yr) and thirteen older (61-71 yr) normotensive, non-endurance-trained women performed both graded and constant-load bouts of leg cycling at the same absolute exercise intensities. Leg blood flow (femoral vein thermodilution), mean arterial pressure (MAP; radial artery), mean femoral venous pressure, cardiac output (acetylene rebreathing), and blood O2 contents were measured. Leg blood flow responses at light workloads (20-40 W) were similar in younger and older women. However, at moderate workloads (50-60 W), leg blood flow responses were significantly attenuated in older women. MAP was 20-25 mmHg higher (P < 0.01) in the older women across all work intensities, and calculated leg vascular conductance (leg blood flow/estimated leg perfusion pressure) was lower (P < 0.05). Exercise-induced increases in leg arteriovenous O2 difference and O2 extraction were identical between groups (P > 0.6). Leg O2 uptake was tightly correlated with leg blood flow across all workloads in both subject groups (r2 = 0.80). These results suggest the ability of healthy older women to undergo limb vasodilation in response to submaximal exercise is impaired and that the legs are a potentially important contributor to the augmented systemic vascular resistance seen during dynamic exercise in older women. PMID- 12882994 TI - Effect of gender on bone turnover in adult rats during simulated weightlessness. AB - Prologned spaceflight results in bone loss in astronauts, but there is considerable individual variation. The goal of this rat study was to determine whether gender influences bone loss during simulated weightlessness. Six-month old Fisher 344 rats were hindlimb unweighted for 2 wk, after which the proximal tibiae were evaluated by histomorphometry. There were gender differences in tibia length, bone area, cancellous bone architecture, and bone formation. Compared with female rats, male rats had an 11.6% longer tibiae, a 27.8% greater cortical bone area, and a 37.6% greater trabecular separation. Conversely, female rats had greater cortical (316%) and cancellous (145%) bone formation rates, 28.6% more cancellous bone, and 30% greater trabecular number. Hindlimb unweighting resulted in large reductions in periosteal bone formation and mineral apposition rate in both genders. Unweighting also caused cancellous bone loss in both genders; trabecular number was decreased, and trabecular separation was increased. There was, however, no change in trabecular thickness in either gender. These architectural changes in cancellous bone were associated with decreases in bone formation and steady-state mRNA levels for bone matrix proteins and cancellous bone resorption. In conclusion, there are major gender-related differences in bone mass and turnover; however, the bone loss in hindlimb unweighted adult male and female rats appears to be due to similar mechanisms. PMID- 12882995 TI - Left ventricular diastolic filling and systolic function of young and older trained and untrained men. AB - Aging is associated with impaired early diastolic filling; however, the effect of endurance training on resting diastolic function in older subjects is unclear. Heart rate and ventricular loading conditions affect mitral inflow velocities measured by Doppler echocardiography; therefore, tissue Doppler imaging of mitral annular velocity, which is relatively preload independent, was combined with mitral inflow velocity and maximal oxygen consumption (V(o2 max)) in young (20-35 yr) and older (60-80 yr) trained and untrained men to determine whether endurance training is associated with an attenuation of age-associated changes in diastolic filling. As expected, V(o2 max) was higher in trained men (P < 0.01) and lower in older men (P < 0.01). Peak early mitral inflow velocity (E) and early-to-late mitral inflow velocity ratios were lower in older vs. young men (P < 0.01); however, there was no training effect (P > 0.05). Peak early mitral annular velocity (E') was higher and peak late mitral annular velocity (A') was lower in young vs. older men (P < 0.01). A significant interaction effect was found for A', E'/A', and peak systolic mitral annular velocity (S'). Training was associated with lower A' in young and higher A' in older men. S' was greater in trained vs. untrained older men (P < 0.05), but it was similar in trained and untrained young men. These findings suggest that early diastolic filling is not affected by training in older men, and the effect of training on A' and S' is different in young and older men. PMID- 12882996 TI - Forebrain and brain stem neural circuits contribute to altered sympathetic responses to heating in senescent rats. AB - Acute heating in young rats increases visceral sympathetic nerve discharge (SND); however, renal and splanchnic SND responses to hyperthermia are attenuated in senescent compared with young Fischer 344 (F344) rats (Kenney MJ and Fels RJ. Am J Physiol Regul Integr Comp Physiol 283: R513-R520, 2002). Central mechanisms by which aging alters visceral SND responses to heating are unknown. We tested the hypothesis that forebrain neural circuits are involved in suppressing sympathoexcitatory responses to heating in chloralose-anesthetized, senescent F344 rats. Renal and splanchnic SND responses to increased (38 degrees C-41 degrees C) internal temperature were determined in midbrain-transected (MT) and sham-MT young (3-mo-old), mature (12-mo-old), and senescent (24-mo-old) F344 rats and in cervical-transected (CT) and sham-CT senescent rats. Renal SND remained unchanged during heating in MT and sham-MT senescent rats but was increased in CT senescent rats. Splanchnic SND responses to heating were higher in MT vs. sham-MT senescent rats and in CT vs. MT senescent rats. SND responses to heating were similar in MT and sham-MT young and mature rats. Mean arterial pressure (MAP) was increased during heating in MT but not in sham-MT senescent rats, whereas heating induced increases in MAP were higher in sham-MT vs. MT young rats. These data suggest that in senescent rats suppression of splanchnic SND to heating involves forebrain and brain stem neural circuits, whereas renal suppression is mediated solely by brain stem neural circuits. These results support the concept that aging alters the functional organization of pathways regulating SND and arterial blood pressure responses to acute heating. PMID- 12882997 TI - iNOS activity is essential for endothelial stress gene expression protecting against oxidative damage. AB - In endothelial cells, the expression of the inducible nitric oxide synthase (iNOS) and the resulting high-output nitric oxide synthesis have often been assumed as detrimental to endothelial function, but recent publications have demonstrated a protective role resulting from iNOS espression and activity. To address this question, we used antisense-mediated iNOS knockdown during proinflammatory cytokine challenge in primary endothelial cell cultures and studied endothelial function by monitoring the expression of stress defense genes. Using antisense oligonucleotides, we achieved a block of iNOS protein formation, accompanied by a strong decrease in the expression of the protective stress response genes bcl-2, vascular endothelial growth factor, and heme oxygenase-1 (HO-1). Additionally, cells were also maintained in the presence of limited exogenous substrate concentrations during cytokine challenge, thereby mimicking a situation of low serum arginine level during inflammation. Under these conditions, cytokine addition results in full iNOS protein expression with minimal nitric oxide formation, concomitant with a significant reduction in stress response gene expression and susceptibility to cell death induced by reactive oxygen species. Taken together, our data suggest that cytokine-induced endogenous iNOS expression and activity have key functions in increasing endothelial survival and maintaining function. Thus suppression of iNOS expression or limited substrate supply, as has been reported to occur in atherosclerosis patients, appears to significantly contribute to endothelial dysfunction and death during oxidative stress. PMID- 12882998 TI - Hormonal changes in satisfied and dissatisfied shift workers across a shift cycle. AB - Although the literature claims that shift work is harmful, it overlooks the fact that that many shift workers are satisfied and stay healthy. There is little knowledge of the biological mechanisms mediating the differences in susceptibility. The present study compared satisfied and dissatisfied shift workers with respect to major anabolic and catabolic hormones. Forty-two male shift workers, with an extremely rapidly rotating shift schedule, were divided into two groups according to their ratings of satisfaction with their work hours. Morning blood samples were taken during the first and last morning shift in the shift cycle. Serum was analyzed with respect to testosterone, cortisol, and prolactin. Dissatisfied shift workers had lower morning testosterone than satisfied ones, but they did not significantly differ with respect to cortisol or prolactin. Low testosterone levels were, in addition, associated with a greater sleep need, disturbed sleep/wakefulness, and an increased need for recovery after the work period, the latter being the best predictor of testosterone levels. The only change across the shift cycle concerned a significant decrease of morning cortisol at the end of the shift cycle. High morning cortisol was related to having a morning personality and fewer sleep problems before the morning shift. Dissatisfaction with the shift system seems related to lower testosterone levels, which in turn are related to disturbed sleep/wakefulness and increased need for sleep and recovery. Furthermore, morning cortisol was reduced across a shift cycle. It is suggested that reduced testosterone levels may be part of a mechanism of shift work maladjustment. PMID- 12882999 TI - Exercise throughout 6 degrees head-down tilt bed rest preserves thermoregulatory responses. AB - Spaceflight and its bed rest analog [6 degrees head-down tilt (HDT)] decrease plasma and blood volume and aerobic capacity. These responses may be associated with impaired thermoregulatory responses observed during exercise and passive heating after HDT exposure. This project tested the hypothesis that dynamic exercise during 13 days of HDT bed rest preserves thermoregulatory responses. Throughout HDT bed rest, 10 subjects exercised for 90 min/day (75% of pre-HDT maximum heart rate; supine). Before and after HDT bed rest, each subject exercised in the supine position at the same workload in a 28 degrees C room. The internal temperature (Tcore) threshold for the onset of sweating and cutaneous vasodilation, as well as the slope of the relationship between the elevation in Tcore relative to the elevation in sweat rate (SR) and cutaneous vascular conductance (CVC; normalized to local heating maximum), were quantified pre- and post-HDT. Tcore thresholds for the onset of cutaneous vasodilation on the chest and forearm (chest: 36.79 +/- 0.12 to 36.94 +/- 0.13 degrees C, P = 0.28; forearm: 36.76 +/- 0.12 to 36.91 +/- 0.11 degrees C, P = 0.16) and slope of the elevation in CVC relative to Tcore (chest: 77.9 +/- 14.2 to 80.6 +/- 17.2%max/ degrees C; P = 0.75; forearm: 76.3 +/- 11.8 to 67.5 +/- 14.3%max/ degrees C, P = 0.39) were preserved post-HDT. Moreover, the Tcore threshold for the onset of SR (36.66 +/- 0.12 to 36.74 +/- 0.10 degrees C; P = 0.36) and the slope of the relationship between the elevation in SR and the elevation in Tcore (1.23 +/- 0.19 to 1.01 +/- 0.14 mg x cm(-2) x min(-1) x degrees C(-1); P = 0.16) were also maintained. Finally, after HDT bed rest, peak oxygen uptake and plasma and blood volumes were not different relative to pre-HDT bed rest values. These data suggest that dynamic exercise during this short period of HDT bed rest preserves thermoregulatory responses. PMID- 12883000 TI - Visualizing infection of individual influenza viruses. AB - Influenza is a paradigm for understanding viral infections. As an opportunistic pathogen exploiting the cellular endocytic machinery for infection, influenza is also a valuable model system for exploring the cell's constitutive endocytic pathway. We have studied the transport, acidification, and fusion of single influenza viruses in living cells by using real-time fluorescence microscopy and have dissected individual stages of the viral entry pathway. The movement of individual viruses revealed a striking three-stage active transport process that preceded viral fusion with endosomes starting with an actin-dependent movement in the cell periphery, followed by a rapid, dynein-directed translocation to the perinuclear region, and finally an intermittent movement involving both plus- and minus-end-directed microtubule-based motilities in the perinuclear region. Surprisingly, the majority of viruses experience their initial acidification in the perinuclear region immediately following the dynein-directed rapid translocation step. This finding suggests a previously undescribed scenario of the endocytic pathway toward late endosomes: endosome maturation, including initial acidification, largely occurs in the perinuclear region. PMID- 12883001 TI - Identification of iron-activated and -repressed Fur-dependent genes by transcriptome analysis of Neisseria meningitidis group B. AB - Iron is limiting in the human host, and bacterial pathogens respond to this environment by activating genes required for bacterial virulence. Transcriptional regulation in response to iron in Gram-negative bacteria is largely mediated by the ferric uptake regulator protein Fur, which in the presence of iron binds to a specific sequence in the promoter regions of genes under its control and acts as a repressor. Here we describe DNA microarray, computational and in vitro studies to define the Fur regulon in the human pathogen Neisseria meningitidis group B (strain MC58). After iron addition to an iron-depleted bacterial culture, 153 genes were up-regulated and 80 were down-regulated. Only 50% of the iron regulated genes were found to contain Fur-binding consensus sequences in their promoter regions. Forty-two promoter regions were amplified and 32 of these were shown to bind Fur by gel-shift analysis. Among these genes, many of which had never been described before to be Fur-regulated, 10 were up-regulated on iron addition, demonstrating that Fur can also act as a transcriptional activator. Sequence alignment of the Fur-binding regions revealed that the N. meningitidis Fur-box encompasses the highly conserved (NATWAT)3 motif. Cluster analysis was effective in predicting Fur-regulated genes even if computer prediction failed to identify Fur-box-like sequences in their promoter regions. Microarray-generated gene expression profiling appears to be a very effective approach to define new regulons and regulatory pathways in pathogenic bacteria. PMID- 12883002 TI - A four-way junction accelerates hairpin ribozyme folding via a discrete intermediate. AB - The natural form of the hairpin ribozyme comprises two major structural elements: a four-way RNA junction and two internal loops carried by adjacent arms of the junction. The ribozyme folds into its active conformation by an intimate association between the loops, and the efficiency of this process is greatly enhanced by the presence of the junction. We have used single-molecule spectroscopy to show that the natural form fluctuates among three distinct states: the folded state and two additional, rapidly interconverting states (proximal and distal) that are inherited from the junction. The proximal state juxtaposes the two loop elements, thereby increasing the probability of their interaction and thus accelerating folding by nearly three orders of magnitude and allowing the ribozyme to fold rapidly in physiological conditions. Therefore, the hairpin ribozyme exploits the dynamics of the junction to facilitate the formation of the active site from its other elements. Dynamic interplay between structural elements, as we demonstrate for the hairpin ribozyme, may be a general theme for other functional RNA molecules. PMID- 12883003 TI - Cell organization in soft media due to active mechanosensing. AB - Adhering cells actively probe the mechanical properties of their environment and use the resulting information to position and orient themselves. We show that a large body of experimental observations can be consistently explained from one unifying principle, namely that cells strengthen contacts and cytoskeleton in the direction of large effective stiffness. Using linear elasticity theory to model the extracellular environment, we calculate optimal cell organization for several situations of interest and find excellent agreement with experiments for fibroblasts, both on elastic substrates and in collagen gels: cells orient in the direction of external tensile strain; they orient parallel and normal to free and clamped surfaces, respectively; and they interact elastically to form strings. Our method can be applied for rational design of tissue equivalents. Moreover, our results indicate that the concept of contact guidance has to be reevaluated. We also suggest that cell-matrix contacts are up-regulated by large effective stiffness in the environment because, in this way, build-up of force is more efficient. PMID- 12883004 TI - In-stream uptake dampens effects of major forest disturbance on watershed nitrogen export. AB - Between January 4 and 10, 1998, a severe ice storm impacted large areas of northern New York, New England, and eastern Canada. This storm struck the Hubbard Brook Experimental Forest in New Hampshire on January 7-8, 1998, and caused extensive forest crown damage (>30%) in a narrow elevation band (600-740 m) across the south-facing experimental watersheds. Stream water has been collected and chemically analyzed since 1963 in six experimental watersheds at Hubbard Brook Experimental Forest; thus, we were able to examine the effect of this severe natural disturbance on watershed nutrient export and changes in instream nitrate (NO3-) processing. The ice storm caused large increases in watershed export of NO3- for 2 years after the disturbance, but our examination of in stream processing suggests that NO3- losses would have been much more dramatic had there not been an increase in in-stream, nitrogen-processing efficiency after the ice storm. The canopy damage that resulted from the ice storm led to increased light availability and large inputs of woody debris to the stream. We suspect that increases in algal production and storage and processing of terrestrial litter account for the increase in inorganic nitrogen processing in these streams. Our results indicate that, without in-stream processing, export of NO3- from the damaged watersheds would have been 80-140% higher than was observed. These results point to an intriguing negative feedback mechanism whereby the same disturbance that causes watershed NO3- loss may simultaneously lead to increased in-stream retention and transformation. PMID- 12883005 TI - Statistical significance for genomewide studies. AB - With the increase in genomewide experiments and the sequencing of multiple genomes, the analysis of large data sets has become commonplace in biology. It is often the case that thousands of features in a genomewide data set are tested against some null hypothesis, where a number of features are expected to be significant. Here we propose an approach to measuring statistical significance in these genomewide studies based on the concept of the false discovery rate. This approach offers a sensible balance between the number of true and false positives that is automatically calibrated and easily interpreted. In doing so, a measure of statistical significance called the q value is associated with each tested feature. The q value is similar to the well known p value, except it is a measure of significance in terms of the false discovery rate rather than the false positive rate. Our approach avoids a flood of false positive results, while offering a more liberal criterion than what has been used in genome scans for linkage. PMID- 12883006 TI - Activity of the Caenorhabditis elegans UNC-86 POU transcription factor modulates olfactory sensitivity. AB - The activity of transcription factors modulates several neural pathways that mediate complex behaviors. We describe here the role of the POU transcription factor UNC-86 in the olfactory behavior of Caenorhabditis elegans. unc-86-null mutants are defective in response to odor attractants but avoid odor repellents normally. Continuous UNC-86 activity is necessary for maintenance of odortaxis behavior; hyperactivation of UNC-86 by fusion to a VP16 activation domain dramatically enhances sensitivity to odor attractants and promotes odor attractant adaptation. UNC-86 is not expressed in olfactory sensory neurons but is expressed throughout the life of the animal in the AIZ interneurons of the odorsensory pathway. We suggest that UNC-86 transcriptional activity regulates the expression of genes that mediate synaptic properties of AIZ and that hyperactive UNC-86::VP16 may enhance the expression of synaptic components to affect the capacity to analyze and process sensory information. PMID- 12883007 TI - Transcriptional regulatory cascades in development: initial rates, not steady state, determine network kinetics. AB - A model was built to examine the kinetics of regulatory cascades such as occur in developmental gene networks. The model relates occupancy of cis-regulatory target sites to transcriptional initiation rate, and thence to RNA and protein output. The model was used to simulate regulatory cascades in which genes encoding transcription factors are successively activated. Using realistic parameter ranges based on extensive earlier measurements in sea urchin embryos, we find that transitions of regulatory states occur sharply in these simulations, with respect to time or changing transcription factor concentrations. As is often observed in developing systems, the simulated regulatory cascades display a succession of gene activations separated by delays of some hours. The most important causes of this behavior are cooperativity in the assembly of cis regulatory complexes and the high specificity of transcription factors for their target sites. Successive transitions in state occur long in advance of the approach to steady-state levels of the molecules that drive the process. The kinetics of such developmental systems thus depend mainly on the initial output rates of genes activated in response to the advent of new transcription factors. PMID- 12883008 TI - An interacting network of T-box genes directs gene expression and fate in the zebrafish mesoderm. AB - T-box genes encode transcription factors that play critical roles in generating the vertebrate body plan. In many developmental fields, multiple T-box genes are expressed in overlapping domains, establishing broad regions in which different combinations of T-box genes are coexpressed. Here we demonstrate that three T-box genes expressed in the zebrafish mesoderm, no tail, spadetail, and tbx6, operate as a network of interacting genes to regulate region-specific gene expression and developmental fate. Loss-of-function and gain-of-function genetic analyses reveal three kinds of interactions among the T-box genes: combinatorial interactions that generate new regulatory functions, additive contributions to common developmental pathways, and competitive antagonism governing downstream gene expression. We propose that T-box genes, like Hox genes, often function within gene networks comprised of related family members. PMID- 12883009 TI - A constitutively activated mutant of human soluble guanylyl cyclase (sGC): implication for the mechanism of sGC activation. AB - Heterodimeric alphabeta soluble guanylyl cyclase (sGC) is a recognized receptor for nitric oxide (NO) and mediates many of its physiological functions. Although it has been clear that the heme moiety coordinated by His-105 of the beta subunit is crucial for mediating the activation of the enzyme by NO, it is not understood whether the heme moiety plays any role in the function of the enzyme in the absence of NO. Here we analyze the effects of biochemical and genetic removal of heme and its reconstitution on the activity of the enzyme. Detergent-induced loss of heme from the wild-type alphabeta enzyme resulted in several-fold activation of the enzyme. This activation was inhibited after hemin reconstitution. A heme deficient mutant alphabetaCys-105 with Cys substituted for His-105 was constitutively active with specific activity approaching the activity of the wild type enzyme activated by NO. However, reconstitution of mutant enzyme with heme and/or DTT treatment significantly inhibited the enzyme. Mutant enzyme reconstituted with ferrous heme was activated by NO and CO alone and showed additive effects between gaseous effectors and the allosteric activator 5 cyclopropyl-2-[1-(2-fluoro-benzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl]-pyrimidin-4 ylamine. We propose that the heme moiety through its coordination with His-105 of the beta subunit acts as an endogenous inhibitor of sGC. Disruption of the heme coordinating bond induced by binding of NO releases the restrictions imposed by this bond and allows the formation of an optimally organized catalytic center in the heterodimer. PMID- 12883010 TI - AthPEX10, a nuclear gene essential for peroxisome and storage organelle formation during Arabidopsis embryogenesis. AB - In yeasts and mammals, PEX10 encodes an integral membrane protein with a C3HC4 RING finger motif in its C-terminal domain and is required for peroxisome biogenesis and matrix protein import. In humans, its dysfunction in peroxisome biogenesis leads to severe Zellweger Syndrome and infantile Refsum disease. Here we show that dysfunction of a homologous gene in Arabidopsis leads to lethality at the heart stage of embryogenesis, impairing the biogenesis of peroxisomes, lipid bodies, and protein bodies. In a T-DNA insertion mutant disrupting the fourth exon of the AthPEX10 gene, ultrastructural analyses fail to detect peroxisomes characteristic for wild-type embryogenesis. Storage triacyl glycerides are not assembled into lipid bodies (oil bodies; oleosomes) surrounded by the phospholipid-protein monolayer membrane. Instead, the dysfunctional monolayer membranes, which derive from the bilayer membrane of the endoplasmic reticulum, accumulate in the cytosol. Concomitantly the transfer of the storage proteins from their site of synthesis at the endoplasmic reticulum to the vacuoles is disturbed. The mutant can be rescued by transformation with wild-type AthPEX10 cDNA. Transformants of wild-type Hansenula polymorpha cells with the AthPEX10 cDNA did produce the encoded protein without targeting it to peroxisomes. Additionally, the cDNA could not complement a Hansenula pex10 mutant unable to form peroxisomes. The ultrastructural knockout phenotype of AthPEX10p suggests that this protein in Arabidopsis is essential for peroxisome, oleosome, and protein transport vesicle formation. PMID- 12883012 TI - SARS and occupational health in the air. PMID- 12883011 TI - Repression of c-Myc and inhibition of G1 exit in cells conditionally overexpressing p300 that is not dependent on its histone acetyltransferase activity. AB - p300 and cAMP response element binding protein (CREB)-binding protein (CBP) are two highly homologous, conserved transcriptional coactivators, and histone acetyltransferases (HATs) that link chromatin remodeling with transcription. Cell transformation by viral oncogene products such as adenovirus E1A and SV40 large T antigen depends on their ability to inactivate p300 and CBP. To investigate the role of p300 in cell-cycle progression, we constructed stable rat cell lines, which conditionally overexpress p300 from a tetracycline-responsive promoter. When p300 was induced in these cells, serum-stimulated S-phase entry was significantly inhibited. The inhibition of S-phase induction was associated with down-regulation of c-Myc, but not of c-Fos or c-Jun. Simultaneous overexpression of c-Myc and p300 before serum stimulation reversed the inhibition of S-phase induction to a significant level, indicating that the inhibition of c-Myc to a large extent is responsible for the p300 inhibition of G1 exit. Similar studies with stable rat cell lines that overexpress a mutant p300, which lacks the HAT activity, showed that the intrinsic HAT activity of p300 is not required for the negative regulation of c-Myc or G1. These findings, and our previously published results (Kolli, S., Buchmann, A. M., Williams, J., Weitzman, S. & Thimmapaya, B. (2001) Proc. Natl. Acad. Sci. USA 98, 4646-4651), establish an important negative regulatory role for p300 in c-Myc expression that may be important in maintaining the cells in the G0/G1 phase of the cell cycle. PMID- 12883013 TI - Epidemiology, biology, and endocrine disrupters. PMID- 12883014 TI - Risk of hypospadias in relation to maternal occupational exposure to potential endocrine disrupting chemicals. AB - BACKGROUND: Reported rises in the prevalence of hypospadias and other abnormalities of the male reproductive system may be a result of exposure to endocrine disrupting chemicals. AIMS: To analyse the relation between risk of hypospadias and maternal occupation, particularly with regard to exposure to potential endocrine disrupting chemicals (EDCs). METHODS: Data (1980-96) from the National Congenital Anomaly System (NCAS) were used to analyse the proportion of all congenital anomaly cases (n = 35 962) which were notified with hypospadias (n = 3471) by occupational codes (348 individual job titles) and by categories of exposure to potential EDCs from a job exposure matrix. RESULTS: Five individual occupations (of 348) showed nominally statistically significant excesses, none of which had possible or probable exposure to potential EDCs. Odds ratios for "possible" or "probable" compared to "unlikely" exposure to potential EDCs did not show statistically significant increases in any of the EDC categories after adjustment for social class of the mother and father, nor was there evidence of an upward trend in risk with likelihood of exposure. In the 1992-96 time period odds ratios were increased for hairdressers (the largest group exposed to potential EDCs) and for probable exposure to phthalates (of which hairdressers form the largest group) before social class adjustment. CONCLUSIONS: There was little evidence for a relation between risk of hypospadias and maternal occupation or occupational exposure to potential EDCs, but as the exposure classification was necessarily crude, these findings should be interpreted with caution. PMID- 12883015 TI - Associations of lead biomarkers with renal function in Korean lead workers. AB - AIMS: To compare associations of lead biomarkers with renal function in current and former lead workers. METHODS: Cross sectional analysis of first year results from a longitudinal study of 803 lead workers and 135 controls in South Korea. Clinical renal function was assessed by blood urea nitrogen (BUN), serum creatinine, and measured and calculated creatinine clearance. Urinary N-acetyl beta-D-glucosaminidase (NAG) and retinol-binding protein were also measured. RESULTS: Mean (SD) tibia lead, blood lead, and DMSA chelatable lead levels in lead workers were 37.2 (40.4) micro g/g bone mineral, 32.0 (15.0) micro g/dl, and 767.8 (862.1) micro g/g creatinine, respectively. Higher lead measures were associated with worse renal function in 16/42 models. When influential outliers were removed, higher lead measures remained associated with worse renal function in nine models. An additional five associations were in the opposite direction. Effect modification by age was observed. In 3/16 models, associations between higher lead measures and worse clinical renal function in participants in the oldest age tertile were significantly different from associations in those in the youngest age tertile which were in the opposite direction. Mean urinary cadmium (CdU) was 1.1 micro g/g creatinine (n = 191). Higher CdU levels were associated with higher NAG. CONCLUSIONS: These data suggest that lead has an adverse effect on renal function in the moderate dose range, particularly in older workers. Associations between higher lead measures and lower BUN and serum creatinine and higher creatinine clearances may represent lead induced hyperfiltration. Environmental cadmium may also have an adverse renal impact, at least on NAG. PMID- 12883016 TI - Municipal waste incinerators: air and biological monitoring of workers for exposure to particles, metals, and organic compounds. AB - AIMS: To evaluate occupational exposure to toxic pollutants at municipal waste incinerators (MWIs). METHODS: Twenty nine male subjects working near the furnaces in two MWIs, and 17 subjects not occupationally exposed to combustion generated pollutants were studied. Individual air samples were taken throughout the shift; urine samples were collected before and after. Stationary air samples were taken near potential sources of emission. RESULTS: Occupational exposure did not result in the infringement of any occupational threshold limit value. Atmospheric exposure levels to particles and metals were 10-100 times higher in MWIs than at the control site. The main sources were cleaning operations for particles, and residue transfer and disposal operations for metals. MWI workers were not exposed to higher levels of polycyclic aromatic hydrocarbons than workers who are routinely in contact with vehicle exhaust. The air concentrations of volatile organic compounds and aldehydes were low and did not appear to pose any significant threat to human health. Only the measurement of chlorinated hydrocarbon levels would seem to be a reliable marker for the combustion of plastics. Urine metal levels were significantly higher at plant 1 than at plant 2 because of high levels of pollutants emanating from one old furnace. CONCLUSION: While biological monitoring is an easy way of acquiring data on long term personal exposure, air monitoring remains the only method that makes it possible to identify the primary sources of pollutant emission which need to be controlled if occupational exposure and environmental pollution are to be reduced. PMID- 12883017 TI - Occupations with increased risk of pancreatic cancer in the Swedish population. AB - AIMS: To identify occupations with increased risk of pancreatic cancer in the Swedish population gainfully employed in 1970 over the period 1971-89. METHODS: The base population was made up of Swedish men (1 779 646) and Swedish women (1 101 669) gainfully employed at the time of the 1970 census and were still alive and over age 24 on 1 January 1971. Information was drawn from two data sets: the Swedish cancer environment register and a background population register. After 19 years of follow up, 4420 men and 2143 women were diagnosed with histologically confirmed incident pancreatic adenocarcinoma. Log linear Poisson models were fitted, allowing for geographical area and town size. Risk estimators were also calculated for workers reporting the same occupation in 1960 and 1970. RESULTS: Among women, a statistically significant risk excess of pancreatic cancer was observed for "educational methods advisors", "librarian, archivist, curator", "motor vehicle driver", "typographer, lithographer", "purser, steward, stewardess", "other housekeeping and related workers", and the groups of occupations of "electrical, electronic, and related" and "glass, pottery, and tile workers". Men showed a higher incidence of pancreatic cancer among "technical assistants", "travelling agents", "other metal processing workers", "baker and pastry cook", "docker and freight handler", and "waiters". CONCLUSIONS: This study does not indicate that occupational factors play an important role in the aetiology of pancreatic cancer in Sweden. Few occupations were at increased risk of pancreatic cancer in both men and women, and the associations observed are in accordance with some previous studies from Western countries. PMID- 12883018 TI - Occupational exposure to electromagnetic fields and acute leukaemia: analysis of a case-control study. AB - AIMS: To investigate whether the risk of acute leukaemia among adults is associated with occupational exposure to electromagnetic fields. METHODS: Probable occupational exposure to electromagnetic fields at higher than typical residential levels was investigated among 764 patients diagnosed with acute leukaemia during 1991-96 and 1510 sex and age matched controls. A job exposure matrix was applied to the self reported employment histories to determine whether or not a subject was exposed to electromagnetic fields. Risks were assessed using conditional logistic regression for a matched analysis. RESULTS: Study subjects considered probably ever exposed to electromagnetic fields at work were not at increased risk of acute leukaemia compared to those considered never exposed. Generally, no associations were observed on stratification by sex, leukaemia subtype, number of years since exposure stopped, or occupation; there was no evidence of a dose-response effect using increasing number of years exposed. However, relative to women considered never exposed, a significant excess of acute lymphoblastic leukaemia was observed among women probably exposed to electromagnetic fields at work that remained increased irrespective of time prior to diagnosis or job ever held. CONCLUSION: This large population based case control study found little evidence to support an association between occupational exposure to electromagnetic fields and acute leukaemia. While an excess of acute lymphoblastic leukaemia among women was observed, it is unlikely that occupational exposure to electromagnetic fields was responsible, given that increased risks remained during periods when exposure above background levels was improbable. PMID- 12883019 TI - Occupational exposures and lung cancer in New Caledonia. AB - AIMS: To study the associations between occupational exposures and the risk of lung cancer in New Caledonia. METHODS: All cases diagnosed between January 1993 and December 1995 (228 lung cancers) and 305 population controls were included. Detailed information on lifetime job history, smoking, and other potential risk factors was collected by interview. Occupational exposures were assessed from the questionnaires by an industrial hygienist, without knowledge of case-control status. RESULTS: No significant association was found with exposures related to nickel mining and refining, the main industrial activity in the territory. Among men, an excess risk of lung cancer was found for bus and truck drivers. Increased risks were also observed in men with the highest level of cumulative exposure to cleaning products and inorganic fertilisers. Exposure to field dust was associated with lung cancer risk in both sexes, and risk increased with cumulative exposure level. In some areas tremolite asbestos derived from local outcroppings was used as a whitewash. The association between exposure to field dust and lung cancer was limited to men and women exposed to this whitewash-that is, living in areas where the soil may contain tremolite. CONCLUSION: This study shows several associations between occupational exposures and lung cancer. The findings suggest that exposure to tremolite fibres from cultivated fields may increase the risk of lung cancer in New Caledonia. PMID- 12883020 TI - Chromium (VI) induced oxidative damage to DNA: increase of urinary 8 hydroxydeoxyguanosine concentrations (8-OHdG) among electroplating workers. AB - AIMS: To investigate the concentration of urinary 8-hydroxydeoxyguanosine (8 OHdG) among electroplating workers in Taiwan. METHODS: Fifty workers were selected from five chromium (Cr) electroplating plants in central Taiwan. The 20 control subjects were office workers with no previous exposure to Cr. Urinary 8 OHdG concentrations were determined using high performance liquid chromatography with electrochemical detection. RESULTS: Urinary 8-OHdG concentrations among Cr workers (1149.5 pmol/kg/day) were higher than those in the control group (730.2 pmol/kg/day). There was a positive correlation between urinary 8-OHdG concentrations and urinary Cr concentration (r = 0.447, p < 0.01), and urinary 8 OHdG correlated positively with airborne Cr concentration (r = 0.285). Using multiple regression analysis, the factors that affected urinary 8-OHdG concentrations were alcohol, the common cold, and high urinary Cr concentration. There was a high correlation of urinary 8-OHdG with both smoking and drinking, but multiple regression analysis showed that smoking was not a significant factor. Age and gender were also non-significant factors. CONCLUSION: 8-OHdG, which is an indicator of oxidative DNA damage, was a sensitive biomarker for Cr exposure. PMID- 12883021 TI - Wastewater exposure and health--a comparative study of two occupational groups. AB - AIMS: To investigate whether wastewater workers are at increased risk of developing cancer. METHODS: Two cohorts of workers employed by the City of Copenhagen, 591 wastewater workers and 1545 water supply workers (controls), were followed from 1965 until 1998. These two cohorts were compared in terms of cause specific mortality and cancer incidence. RESULTS: The wastewater workers' mortality exceeded that of the controls (relative risk (RR) = 1.25, 95% CI: 1.03 to 1.51). A similar small excess was seen for cancer incidence (RR = 1.27, 95% CI: 0.97 to 1.67). Though rare, there was a strongly increased incidence of primary liver cancer among the wastewater workers (RR = 8.9, 95% CI: 1.5 to 51.5). CONCLUSION: The excess mortality seen among the wastewater workers was smaller than originally feared. It may partly have been due to their occupational exposure, and for preventive purposes, exposure to wastewater and sludge should be minimised. The possibility that sewage exposure confers an increased risk of primary liver cancer deserves further investigation. PMID- 12883022 TI - Exposure assessment of monoterpenes and styrene: a comparison of air sampling and biomonitoring. AB - BACKGROUND: Within- and between-worker variance components have seldom been reported for both environmental and biological data collected from the same persons. AIMS: To estimate these variance components and their ratio for air contaminants and urinary metabolites in two different work environments and to predict the attenuation of exposure-response relationships based on these measures. METHODS: Parallel measurements of air and urine were performed among workers exposed to monoterpenes in sawmills (urinary metabolite: verbenol) and styrene in reinforced plastics factories (urinary metabolite: mandelic acid). RESULTS: Among the sawmill workers, variance components of the air and urinary verbenol results were similar; for the reinforced plastics workers the estimated between-worker variance component was greater for styrene in air than mandelic acid in urine. This suggests that attenuation bias would be about equal if air or biological monitoring were employed for monoterpene exposures, but would be greater if urinary mandelic acid were used instead of airborne styrene in an investigation of styrene exposure. CONCLUSIONS: Personal air samplers provide data with similar or superior quality to urinary metabolites as measures of exposure to these monoterpenes in sawmills and styrene in reinforced plastics factories. PMID- 12883023 TI - Estimating the incidence of occupational asthma and rhinitis from laboratory animal allergens in the UK, 1999-2000. AB - We report a survey of the number of persons handling laboratory animals in the UK in 1999-2000. This estimate has allowed us to calculate annual incidence rates of allergic asthma and rhinitis attributable to laboratory animals, insects, and birds using numerators derived from cases reported to a national surveillance scheme (SWORD). The number of exposed employees has almost halved since the early 1980s. From these data, the estimated annual incidence rate of occupational asthma among persons working with small mammals is 1.56/1000 employees. Although reported cases are rare, estimated incidence rates of asthma attributable to birds (23.9/1000/year) and insects (2.35/1000/year) are high. PMID- 12883024 TI - Should computed chest tomography be recommended in the medical certification of professional divers? A report of three cases with pulmonary air cysts. AB - Pulmonary barotrauma (PBT) is a recognised risk of compressed gas diving. Any reason that causes air trapping in the lung during ascent may cause PBT by increasing intrapulmonary pressure. Chest x ray examination is mandatory for medical certification of the professional divers in many countries, but pulmonary air trapping lesions such as an air cyst in the lungs cannot always be detected by plain chest x ray examination. Computed tomography (CT) is a reliable, but expensive measure for detecting pulmonary abnormalities in divers. Three cases with pulmonary air cysts are reported in which air cysts were invisible on the x ray pictures, but well defined by CT. It is impractical and not cost effective to perform CT for medical certification of all divers, but it can be an option to recommend CT once during the initial examination of the candidates for professional diving, especially if there is a history of predisposing factors, such as smoking or pulmonary infections. PMID- 12883025 TI - Dermatological powder as hidden cause of occupational allergy due to casein: a case report. AB - We report a case of occupational asthma due to hidden casein, contained in a dermatological powder. A 44 year old nurse reported the recent onset of rhinitis and asthma, clearly related to the work environment. Skin tests for food allergens revealed a positivity for milk proteins. The source of dispersed milk proteins in the work environment was found to be a dermatological formulation of an inert refreshing powder, used to treat excoriated areas in the elderly. Skin prick tests confirmed the positivity to casein and to the commercial product. PMID- 12883026 TI - Earlier study on asbestos workers, ILO scores, and oxygenation more comprehensive. PMID- 12883027 TI - Oxidative stress: its role in air pollution and adverse health effects. PMID- 12883028 TI - Ischaemic heart disease among livestock and agricultural workers. AB - BACKGROUND: Agricultural workers are exposed to organic dust containing endotoxins, mites, and moulds. AIMS: To compare the occurrence of ischaemic heart disease (IHD) among male and female livestock and agricultural workers with gainfully employed men and women in Sweden. METHODS: Male and female livestock and agricultural workers were identified in the Swedish National Censuses of 1970 and 1990 and were followed until the end of 1995. The IHD mortality among the livestock and agricultural workers was compared with that of gainfully employed men and women. Information of smoking habits was gathered from a previous national survey. RESULTS: Male as well as female livestock workers had slightly higher standardised mortality ratios (SMR) regarding IHD compared with all gainfully employed men and women in Sweden. The SMR for male workers was 1.06 (95% CI 0.95 to 1.18). The SMR for female workers was 1.10 (95% CI 0.98 to 1.23). Agricultural workers had lower SMRs. Adjustments for smoking habits would further increase the SMRs by about 9% in male workers and about 5% in female workers. CONCLUSION: The present data suggest a slightly increased risk for IHD among both male and female livestock workers, which may be the result of organic dust exposure. PMID- 12883029 TI - Respiratory effects of sulphur dioxide: a hierarchical multicity analysis in the APHEA 2 study. AB - BACKGROUND: Sulphur dioxide (SO(2)) was associated with hospital admissions for asthma in children in the original APHEA study, but not with other respiratory admissions. AIMS: To assess the association between daily levels of SO(2) and daily levels of respiratory admissions in a larger and more recent study. METHODS: Time series of daily counts of hospital emergency admissions were constructed for asthma at ages 0-14 years and 15-64 years, COPD and asthma, and all respiratory admissions at ages 65+ years in the cities of Birmingham, London, Milan, Paris, Rome, Stockholm, and in the Netherlands for periods of varying duration between the years 1988 and 1997. A two stage hierarchical modelling approach was used. In the first stage generalised additive Poisson regression models were fitted in each city controlling for weather and season. These results were then combined across cities in a second stage ecological regression that looked at potential effect modifiers. RESULTS: For an increase of 10 micro g/m(3) of SO(2) the daily number of admissions for asthma in children increased 1.3% (95% CI 0.4% to 2.2%). Effect modification among cities by levels of other air pollutants or temperature was not found. The SO(2) effect disappeared after controlling for PM(10) or CO, but correlation among these pollutants was very high. Other respiratory admissions were not associated with SO(2). CONCLUSION: SO(2) is associated with asthma admissions in children, indicating that reduction in current air pollution levels could lead to a decrease in the number of asthma admissions in children in Europe. PMID- 12883030 TI - Animal production and wheeze in the Agricultural Health Study: interactions with atopy, asthma, and smoking. AB - BACKGROUND: Exposure to animals, their feeds, and by-products contribute to respiratory symptoms among farmers. AIMS: To investigate the role of animal exposures and wheeze, and to assess whether their impact differs among susceptible subgroups, including atopics, asthmatics, and smokers. METHODS: Using the Agricultural Health Study, a cohort of pesticide applicators in Iowa and North Carolina enrolled in 1994-97, wheeze associated with animal production was evaluated and interactions among susceptible subgroups assessed. Logistic regression models were used to examine risk factors for wheeze in the past year among 20 468 farmers. RESULTS: Individuals raising animals requiring direct contact had the highest odds ratios (OR) for wheeze (OR(dairy) = 1.26; OR(eggs) = 1.70). A significant dose response was observed for both the number of poultry and the number of livestock on the farm. Farmers who performed veterinary procedures on a daily basis had an OR of 1.51. The odds of wheeze associated with poultry production was greater among atopic than non-atopic individuals. Milking cows daily increased the odds of wheeze in all individuals, with the largest association observed among atopic asthmatic individuals. The impact of dairy, poultry, and egg production varied among smoking groups. Past smokers had the highest odds ratios, followed by never smokers, and then current smokers. The OR(eggs) was 2.88 among past smokers but only 1.46 for never smokers. The OR(eggs) for current smokers of 0.80 might reflect self selection of exposure among smokers. CONCLUSIONS: Results are consistent with animal production and respiratory symptoms, and suggest that subgroups may respond differently to exposure. PMID- 12883031 TI - Mutations in alpha- and beta-tubulin that stabilize microtubules and confer resistance to colcemid and vinblastine. AB - Single-step selections were used to obtain Chinese hamster ovary cell lines resistant to Colcemid and vinblastine. Verapamil was included in the selections to circumvent the isolation of cells with P-glycoprotein-mediated multidrug resistance and thereby enrich for cells with tubulin alterations. The isolated cell lines were 2-fold resistant to the selecting drug, exhibited cross resistance to other drugs that inhibit microtubule assembly, and had enhanced sensitivity to the microtubule-stabilizing drug paclitaxel. The concomitant resistance to microtubule-destabilizing drugs and enhanced sensitivity to paclitaxel suggested that these cell lines have changes in microtubule assembly. Consistent with this interpretation, drug-resistant cell lines exhibited altered alpha- or beta-tubulin mobility on two-dimensional gels and higher levels (47 54%) of assembled tubulin compared with wild-type (39%) or paclitaxel-resistant cells (25%). Some drug-resistant cells also had bundled microtubules as judged by immunofluorescence. Genomic sequencing of 11 drug-resistant cell lines predicted five different alterations (D45Y, C211F, D224N, S234N, and K350N) in beta-tubulin and four different alterations (H283Y, E55K, A383V, and R390C) in alpha-tubulin. The amino acid substitutions are dispersed on the primary and tertiary structures of tubulin and, together with the other mutant properties, argue against a mechanism involving changes in drug binding. Rather, we propose that the alterations in alpha- and beta-tubulin increase microtubule stability by promoting longitudinal interdimer and intradimer interactions and/or lateral interactions between protofilaments. This enhanced stability of microtubules increases their resistance to drugs that inhibit assembly. PMID- 12883032 TI - Alchemix: a novel alkylating anthraquinone with potent activity against anthracycline- and cisplatin-resistant ovarian cancer. AB - Chloroethylaminoanthraquinones are described with intercalating and alkylating capacity that potentially covalently cross-link topoisomerase II (topo II) to DNA. These compounds have potent cytotoxic activity (IC(50) = 0.9-7.6 nM) against the A2780 human ovarian carcinoma cell line. Hydroxyethylaminoanthraquinones also reported in this paper have similar IC(50) values (0.7-1.7 nM) in the same cell line. Alchemix (ZP281M, 1-(2-[N,N-bis(2-chloroethyl)amino]ethylamino)-4-(2-[N,N (dimethyl)amino]ethylamino)-5,8-dihydroxy-9,10-anthracenedione), an alkylating anthraquinone, retains excellent antitumor activity in Adriamycin-resistant (2780AD) and cisplatin-resistant (2780/cp70) cell lines in vitro and in vivo. This indicates that Alchemix can evade both P-glycoprotein efflux pump and DNA mismatch repair-mediated resistance. In treated cells, Alchemix was shown to preferentially induce drug-stabilized covalent bound topo IIalpha-DNA complexes over topo IIbeta-DNA complexes. PMID- 12883033 TI - SU5416 selectively impairs angiogenesis to induce prostate cancer-specific apoptosis. AB - We have previously demonstrated the differential expression in tumor-associated blood vessels of two vascular endothelial growth factor receptors (VEGFRs), VEGFR1 and VEGFR2, during initiation and progression of prostate cancer in the genetically engineered transgenic adenocarcinoma of the mouse prostate (TRAMP) mouse model. In our "progression switch" model, expression of VEGFR1 is associated with early and more differentiated disease, whereas expression of VEGFR2 is associated with advanced and more poorly differentiated disease. To test the hypothesis that stage-specific inhibition of vascular endothelial growth factor signaling could be used as therapy for autochthonous prostate cancer, we initiated a preclinical trial with SU5416, a potent antiangiogenic small molecule inhibitor of VEGFR associated tyrosine kinase activity. In our early intervention trial, administration of SU5416 to TRAMP mice did not appear to influence angiogenesis or tumor progression between 10 and 16 weeks of age, a time corresponding to high levels of VEGFR1 expression. In our late intervention trial, however, we observed a significant decrease in tumor-associated mean vessel density, increased apoptotic index, and pronounced regions of cell death when SU5416 was administered to TRAMP mice between 16 and 22 weeks of age, a time corresponding to high levels of VEGFR2 expression. These results clearly demonstrate that therapy directed specifically against the VEGFR signaling axis can dramatically impair angiogenesis and induce apoptosis of autochthonous spontaneous and progressive prostate cancer. PMID- 12883034 TI - Antiangiogenic tetrathiomolybdate enhances the efficacy of doxorubicin against breast carcinoma. AB - Constitutive activation of nuclear factor kappaB is implicated to be a critical survival mechanism used by carcinoma cells to escape apoptosis. Tetrathiomolybdate (TM), a novel copper chelator, exhibits potent antiangiogenic properties, in part, through suppression of the nuclear factor kappaB signaling cascade. In this study, we determined whether TM enhances doxorubicin-induced apoptosis in SUM149 inflammatory breast carcinoma cells. Apoptosis was not observed in these cells after TM treatment. Moreover, SUM149 cells were relatively resistant to doxorubicin-induced apoptosis ranging from 9.9 +/- 2.9% to 21.5 +/- 2.0% apoptotic cells for 0.1 to 2.5 micro M doxorubicin treatment. A greater-than-additive increase (33.8 +/- 4.6%, 57.5 +/- 5.2%, or 83.7 +/- 1.0% apoptosis with TM and 0.1, 0.5, or 2.5 micro M doxorubicin, respectively) in apoptosis was observed in cells treated with the combination therapy of TM and doxorubicin. In SUM149 xenografts, TM and doxorubicin significantly retarded tumor growth in comparison with either agent administered alone (P < 0.03). Tumor cell apoptosis in the combination therapy-treated mice was 113.3 +/- 20% greater than that in TM-treated mice and 52.4 +/- 14.3% greater than that in doxorubicin treated mice. These results suggest that TM may enhance the rate of pathological complete response when used in combination with an anthracycline in neoadjuvant therapy of breast carcinoma. PMID- 12883035 TI - MDA-7 (interleukin-24) inhibits the proliferation of renal carcinoma cells and interacts with free radicals to promote cell death and loss of reproductive capacity. AB - The median survival of metastatic renal cell carcinoma (RCC) is 12 months, and the majority of treatment options are palliative. MDA-7 (interleukin-24), when expressed via a recombinant replication defective adenovirus, Ad.mda-7, has profound antiproliferative and cytotoxic effects in a wide variety of tumor cells but not in nontransformed cells. The studies in this study examined the impact of MDA-7 on RCC proliferation and survival. RCC lines (A498 and UOK121N), but not primary renal epithelial cells, were resistant to adenoviral infection that correlated with a lack of coxsackievirus and adenovirus receptor expression. Additional studies were performed using purified preparations of bacterially synthesized glutathione S-transferase (GST)-MDA-7 protein. GST-MDA-7, but not GST, caused a dose-dependent inhibition of RCC proliferation but not of primary renal epithelial cells. Clinically achievable concentrations of the novel therapeutic agent arsenic trioxide (0.5-1 micro M) were found to have little effect on RCC growth. However, the combination of GST-MDA-7 and arsenic trioxide resulted in a greater than additive reduction in cell growth that correlated with a large increase in tumor cell death. The free radical scavenger N-acetyl cysteine abolished the potentiating effect of arsenic trioxide. Although pro caspase 3, poly(ADP-ribose) polymerase, and Bcl-(XL) levels, as well as nucleosomal DNA integrity, were reduced by combined treatment, cell killing was predominantly nonapoptotic. Combined treatment of RCC lines with GST-MDA-7 and arsenic trioxide also resulted in a substantial reduction in clonogenic survival compared with either treatment individually. Collectively, these findings demonstrate that MDA-7 protein, in combination with agents that generate free radicals, may have potential in the treatment of RCC. PMID- 12883036 TI - Enhancement of platinum-induced cytotoxicity by O6-benzylguanine. AB - O(6)-Benzylguanine (O(6)-BG), a potent inactivator of the DNA repair protein O(6) alkylguanine-DNA alkyltransferase (AGT), is presently in clinical trials combined with alkylating agents that modify the O(6) position of DNA guanine residues, i.e., 1,3-bis(2-chloroethyl)-1-nitrosourea and temozolomide. Previous work demonstrated that O(6)-BG also enhances the cytotoxicity of cyclophosphamide, ifosfamide, and nitrogen mustards in Chinese hamster ovary cells. We have extended this study to include other clinically relevant agents that form interstrand and intrastrand cross-links including cisplatin and carboplatin. Pretreatment of a series of head and neck tumor cell lines (i.e., SQ20b, JSQ3, SCC25, SCC35, and SCC61), Chinese hamster ovary cells, and HT29 human colon tumor cells with O(6)-BG (100 micro M for 2 h before treatment and 2 h during treatment) resulted in a 2-fold decrease in the ED(50) of cisplatin and a concomitant increase in the percentage of cells undergoing apoptosis. The enhancement was independent of AGT activity. Similar enhancement was observed with carboplatin, but no enhancement was seen in AGT-deficient cell lines with radiation or temozolomide, demonstrating the dependence of the effect on bifunctional, cross-linking agents. Furthermore, levels of platinum on DNA after treatment with cisplatin increased 1.4-fold in SQ20b cells and 4.5-fold in JSQ3 cells immediately after treatment with O(6)-BG plus cisplatin and remained elevated for 48 h. Consistent with greater cytotoxicity and apoptosis is the approximately 2-fold higher amount of DNA damage when cells are treated with O(6) BG plus cisplatin compared with cisplatin alone. Modulation of cisplatin therapy with O(6)-BG might improve the prognosis of patients with head and neck, ovarian, testicular, or lung cancer who are treated with this drug. PMID- 12883037 TI - Inhibition of mitogen-activated protein kinase/extracellular signal-regulated kinase kinase enhances chemotherapeutic effects on H460 human non-small cell lung cancer cells through activation of apoptosis. AB - The effects of Dox (Dox), paclitaxel (Taxol), and serum starvation on the regulation of XIAP (X-linked inhibitor of apoptosis), Bcl-2 phosphorylation, and apoptosis were evaluated in human H460 non-small cell lung cancer cells. Protein kinases that responded to these treatments as prosurvival elements in signal transduction were identified by simultaneously screening phosphorylation of protein kinases in H460 cells cultured in serum-free medium or treated with Dox. We demonstrated that Dox and Taxol induced apoptosis through down-regulation of XIAP and phosphorylation of Bcl-2 in a concentration-dependent manner without changing expression of Bcl-xL in H460 cells. These effects were paralleled by activation of caspase-3 and cleavage of poly(ADP-ribose) polymerase protein. We identified that serum starvation and Dox reduced phosphorylation of mitogen activated protein kinase/extracellular signal-regulated kinase (ERK) kinase (MEK), protein kinase C (PKC) alpha/beta and c-Jun NH(2)-terminal kinase. The MEK specific inhibitor U0126 or PKC inhibitor staurosporine (STP) also down-regulated XIAP expression and induced apoptosis. Thus, our data suggest that apoptosis and down-regulation of XIAP induced by Dox exposure or serum starvation may be mediated through inactivation of the MEK/ERK and PKCalpha/beta pathways. In support of this we demonstrated that the cytotoxic effects of Dox when combined with U0126 or STP were enhanced, i.e., synergistic cytotoxic activities were demonstrated. The synergistic interaction of U0126 or STP with Dox was sequence- and concentration-dependent. PMID- 12883038 TI - The DNA minor groove-alkylating cyclopropylpyrroloindole drugs adozelesin and bizelesin induce different DNA damage response pathways in human colon carcinoma HCT116 cells. AB - As members of the cyclopropylpyrroloindole family, adozelesin and bizelesin cause genomic DNA lesions by alkylating DNA. Adozelesin induces single-strand DNA lesions, whereas bizelesin induces both single-strand lesions and double-strand DNA cross-links. At equivalent cytotoxic concentrations, these agents caused different biological responses. Low adozelesin concentrations (e.g., 0.5 nM) induced a transient S-phase block and cell cycle arrest in G(2)-M, as well as increased induction of p53 and p21, whereas a high drug concentration (e.g., 2.5 nM) caused apoptosis but no p21 induction. In contrast, both low and high bizelesin concentrations enhanced p53 and p21 induction and triggered G(2)-M cell cycle arrest and eventual senescence without significant apoptotic cell death. However, in cells lacking p21, bizelesin, as well as adozelesin, triggered apoptosis, indicating that p21 was crucial to sustained bizelesin-induced G(2)-M arrest. Thus, despite similar abilities to alkylate DNA, the chemotherapeutic agents adozelesin and bizelesin caused a decrease in HCT116 tumor cell proliferation by different pathways (i.e., adozelesin induced apoptosis, and bizelesin induced senescence). PMID- 12883039 TI - Sequence specificity of adriamycin-DNA adducts in human tumor cells. AB - The anticancer anthracycline compound Adriamycin is a known topoisomerase II inhibitor but is also capable of exerting other cellular consequences. After intercalation, Adriamycin can form covalent adducts with DNA, and the magnitude of these adducts appears to be limited by the cellular availability of formaldehyde. Adducts produced by Adriamycin in the presence of formaldehyde have been well characterized in cell-free systems but not in cells. In this study, we show that when Adriamycin is used in conjunction with the formaldehyde-releasing prodrug AN-9 in IMR-32 tumor cells, this allows the formation of sufficiently high levels of adducts in genomic DNA to enable detection of their DNA sequence specificity for the first time. The 340-bp alpha-satellite EcoRI repeat sequence was isolated from drug-treated cells and digested with lambda-exonuclease to determine adduct sites at which exonuclease digestion was blocked. The Adriamycin adducts were formed predominantly at 5'-GC and GG sequences and unstable with respect to elevated temperatures and extended times at 37 degrees C. The use of three anthracycline derivatives lacking a 3'amino group demonstrated that this amino portion is critical for the formation of anthracycline adducts in cells. The structure of these drug-DNA adducts can therefore be considered to be identical to the Adriamycin adducts, which have been characterized rigorously in cell-free systems by X-ray crystallography, two-dimensional nuclear magnetic resonance, and mass spectrometry. PMID- 12883040 TI - Gene expression profiles in a panel of childhood leukemia cell lines mirror critical features of the disease. AB - The development of new drugs against cancer requires established cell lines. They are needed for in vitro studies to identify candidate drugs and in xenograft models to measure drug efficacy in vivo. Specific criteria need to be fulfilled by cell lines used in the evaluation of potential novel therapeutic agents. It is imperative that they display the features of the particular cancer under investigation. Given the documented heterogeneity of cancers, relevant subtypes need to be represented. In this study, we have examined these aspects for pediatric acute lymphoblastic leukemia. A panel of 13 leukemia cell lines recently established in our laboratory was analyzed. We used cDNA microarrays to define the gene expression profiles and compared the data with immunophenotyping and cytogenetic analyses. The expression profiles obtained showed excellent concordance with corresponding protein levels. Importantly, the panel of lines displayed the critical genetic features identified in clinically important acute lymphoblastic leukemia subtypes in childhood leukemia patients. PMID- 12883041 TI - Binarization of microarray data on the basis of a mixture model. AB - Although gathered as continuous data, expression measurements from gene microarrays may be quantized before downstream analysis and modeling. This is especially true for modeling gene prediction and genetic regulatory networks. Coarse quantization results in lower computational requirements, lower data requirements for model inference, and easier conceptualization. This paper proposes a mixture model for binarization. For each gene, the model, composed of a sum of two distributions, is fit to expression data for that gene, and data points are binarized according to the model. The mixture model is based on the assumption of multiplicative up-regulation. The proposed method is compared with mean and median binarization by comparing classification performance based on the binary data from the different methods. Classification is performed for simulated data generated from a microarray model studied previously and for cancer data arising from two studies involving hereditary breast cancer and small, round blue cell tumors of childhood. PMID- 12883042 TI - Pharmacogenetics of ATP-binding cassette transporters in cancer and chemotherapy. AB - The ATP-binding cassette (ABC) transporters belong to the largest known transporter gene family and translocate a variety of substrates including chemotherapy agents. ABC multidrug transporter expression has been implicated in tumor cell resistance to anticancer therapy, altered disposition of chemotherapy drugs, and associated chemotherapy toxicity. More recently, genetic heterogeneity has been described in a number of the ABC transporter genes, including ABC transporters that contribute to the pharmacokinetics and/or pharmacodynamics of chemotherapy drugs. The role of these transporters and their naturally occurring genetic polymorphisms in cancer and chemotherapy is reviewed. PMID- 12883043 TI - Identification of three previously unknown in vivo protein phosphorylation sites in thylakoid membranes of Arabidopsis thaliana. AB - The proteins in plant photosynthetic thylakoid membranes undergo light-induced phosphorylation, but only a few phosphoproteins have been characterized. To access the unknown sites of in vivo protein phosphorylation the thylakoid membranes were isolated from Arabidopsis thaliana grown in normal light, and the surface-exposed peptides were cleaved from the membranes by trypsin. The peptides were methylated and subjected to immobilized metal affinity chromatography, and the enriched phosphopeptides were sequenced using tandem nanospray quadrupole time-of-flight mass spectrometry. Three new phosphopeptides were revealed in addition to the five known phosphorylation sites in photosystem II proteins. All phosphopeptides are found phosphorylated at threonine residues implementing a strict threonine specificity of the thylakoid kinases. For the first time protein phosphorylation is found in photosystem I. The phosphorylation site is localized to the first threonine in the N terminus of PsaD protein that assists in the electron transfer from photosystem I to ferredoxin. A new phosphorylation site is also revealed in the acetylated N terminus of the minor chlorophyll a-binding protein CP29. The third novel phosphopeptide, composed of 25 amino acids, belongs to a nuclear encoded protein annotated as "expressed protein" in the Arabidopsis database. The protein precursor has a chloroplast-targeting peptide followed by the mature protein with two transmembrane helices and a molecular mass of 14 kDa. This previously uncharacterized protein is named thylakoid membrane phosphoprotein of 14 kDa (TMP14). The finding of the novel phosphoproteins extends involvement of the redox-regulated protein phosphorylation in photosynthetic membranes beyond the photosystem II and its light-harvesting antennae. PMID- 12883044 TI - Characterization of a subfamily of beetle odorant-binding proteins found in hemolymph. AB - In insects, hydrophobic odorants are transported through the sensillar lymph to receptors on sensory neurons by odorant-binding proteins (OBPs). The beetle Tenebrio molitor, which is a pest of stored grain products, produces a set of 12 14-kDa OBP-like proteins in its hemolymph. The structure of one of these proteins and that of a moth pheromone-binding protein have been solved. Both proteins have at least six alpha-helices with an internal, hydrophobic, ligand-binding pocket, but the beetle OBP lacks one of the disulfide bonds immediately adjacent to this pocket. To explore this difference and to sample isoform diversity, T. molitor hemolymph OBPs were fractionated by size-exclusion chromatography and reversed phase high performance liquid chromatography. Selected fractions were reduced and alkylated, and tryptic peptides were sequenced by tandem mass spectrometry. Partial sequences of 7 different isoforms were obtained and used to clone 9 new cDNAs encoding OBPs with identities from 32 to 99%. The more divergent isoforms have numerous substitutions of hydrophobic residues that presumably alter the shape and specificity of the ligand-binding pocket. These isoforms all lack the same third disulfide bridge and are more similar to one another than to any of the 38 OBPs in Drosophila melanogaster. They have presumably arisen via gene duplication following separation of the major insect orders. PMID- 12883046 TI - Chasing the dollar: why scientists should decline tobacco industry funding. PMID- 12883047 TI - Hazardous effects of tobacco industry funding. PMID- 12883048 TI - Multilevel analytical approaches in social epidemiology: measures of health variation compared with traditional measures of association. PMID- 12883050 TI - Psychosocial factors and public health. PMID- 12883051 TI - Unemployment and suicide. PMID- 12883052 TI - Unemployment and suicidal behaviour. PMID- 12883053 TI - Research on unemployment and suicide. PMID- 12883054 TI - Unemployment and suicide. PMID- 12883055 TI - Advanced glossary on genetic epidemiology. AB - This is the last of a series of glossaries on terms used in genetic epidemiology published by the journal. This glossary covers the most advanced genetic terms, most of which are related to new study designs and laboratory techniques. It provides the reader with examples and references of real studies that applied each of the study designs defined in the glossary. This should help the reader grasp the subtleties of each of these strategies and will allow the reader to research the literature according to their interest. PMID- 12883057 TI - Psychosocial factors and public health: a suitable case for treatment? AB - Adverse psychosocial exposure or "misery" is associated with physical disease. This association may not be causal. Rather it may reflect issues of reverse causation, reporting bias, and confounding by aspects of the material environment typically associated with misery. A non-causal relation will not form the basis of effective public health interventions. This may be why psychosocial interventions have, so far, showed little effect on objective physical health outcomes. This paper reviews evidence for the "psychosocial hypothesis" and suggests strategies for clarifying these issues. It concludes that, although misery is clearly a bad thing as it erodes people's quality of life, there is little evidence that psychosocial factors cause physical disease. In the absence of better evidence, suggestions that psychosocial interventions are needed to improve population physical health, in both absolute and relative terms, seem premature. PMID- 12883060 TI - Smoking cessation services are reducing inequalities. PMID- 12883059 TI - Chasing Ernst L Wynder: 40 years of Philip Morris' efforts to influence a leading scientist. AB - STUDY OBJECTIVE: To highlight strategies used by the Philip Morris tobacco company to try to manipulate the eminent scientist, Dr Ernst Wynder between 1955 and 1995. METHODS: Systematic keyword and opportunistic searching of www.pmdocs.com for formerly internal tobacco industry documents concerning Philip Morris executives and Wynder. Available materials included reports, budget reviews, and correspondence. MAIN RESULTS: The emergence of smoking as a priority issue on the American public health agenda can be largely attributed to Wynder's research and publicity efforts. Philip Morris viewed Wynder as a prestigious scientist whose commitment to the pursuit of reduced harm cigarettes could lend legitimacy to its desire to position itself as a responsible company intent on addressing consumer concerns. Philip Morris courted Wynder with large equipment loans and grants for more than 30 years, and used its public relations agency to sanitise press releases to remove material unacceptable to the company. Wynder consistently failed to acknowledge industry support while routinely acknowledging other funding from the National Cancer Institute and the American Cancer Society. In retrospect, Wynder realised the insidious effect of tobacco industry research support but failed to acknowledge this may have applied to his own association with the industry. CONCLUSIONS: Industry documents reveal a deliberate attempt by Philip Morris to pursue and manipulate Dr Wynder to legitimise their company positions. PMID- 12883061 TI - Social deprivation and the public health risks of community drinking water supplies in New Zealand. AB - STUDY OBJECTIVE: Quantitative evidence linking environmental exposures and social status at sub-national scales is surprisingly limited. This study investigated the public health risks associated with community water supplies in relation to social status in New Zealand. DESIGN: An ecological study using a Geographic Information System (GIS) to compare the grade of community water supplies with an index of social deprivation for small areas. SETTING: New Zealand. PARTICIPANTS: The New Zealand population usually resident in meshblocks (census areas) with a community water supply (70% of the 1996 population of 3.6 million people). MAIN RESULTS: People living in deprived areas are exposed to greater public health risks from community water supplies. In urban areas, the odds of water supplies being high risk were 3.76 times greater for the most deprived decile compared with the least deprived decile (95% CI: 2.95 to 4.78). CONCLUSIONS: It is probable that deprived communities in New Zealand are experiencing a disproportionate burden of adverse health effects as a result of poor water quality. PMID- 12883062 TI - Does an increase of low income families affect child health inequalities? A Swedish case study. AB - STUDY OBJECTIVE: Reduction of health inequalities is a primary public health target in many countries. A change of proportion of low income families might affect child health inequalities. Yet, the importance of family incomes in high income welfare states is not well established. The aim of this study was to investigate the effect of increased percentage of low income families on child health inequalities during an economic recession in Sweden, 1991-1996. DESIGN: Health inequalities for six health indicators were assessed during the period 1991-1996 and during adjacent periods. Relative inequality indices were estimated according to Pamuk and Mackenback. Appraisal of a child's socioeconomic situation was based on social data for the child's residency area. SETTING: The total population of children and adolescents 0-<19 years old living in Stockholm County, Sweden, was studied. Each one year cohort comprised 20 470-25 420 people. MAIN OUTCOME MEASURES: Mortality; rate of low birth weight; days of hospital care for infections, asthma/allergic disorders, and unintentional injuries; and rate of abortions. MAIN RESULTS: Mortality decreased annually by 6.9%. The average relative inequality index for mortality before the recession was 1.40 and was lower during the recession, 1.14. The remaining five health indicators, and the relative inequality index for these indicators, did not differ significantly between the recession years (1991-1996) and adjacent periods. CONCLUSIONS: Relative health inequalities did not change, or decreased, during the recession years. The findings indicate that the connection was weak between child health inequalities and family incomes, within the frame of time and the range of income changes that occurred during the study period. PMID- 12883063 TI - Randomised controlled trial of the effect of evidence based information on women's willingness to participate in cervical cancer screening. AB - STUDY OBJECTIVE: To assess whether providing women with additional information on the pros and cons of screening, compared with information currently offered by the NHS, affects their intention to attend for screening. DESIGN: Randomised controlled trial. Participants were randomly assigned to receive either the control, (based on an NHS Cervical Screening Programme leaflet currently used), or the intervention leaflet (containing additional information on risks and uncertainties). SETTING: Three general practices in Birmingham. PARTICIPANTS: 300 women aged 20 to 64 attending the practices during a one month period. MAIN OUTCOME MEASURES: Intention to attend for screening. MAIN RESULTS: 283 women (94.3%) completed the study. Fewer women in the intervention (79%) than the control group (88%) expressed intention to have screening after reading the information leaflet (difference between groups 9.2%, 95% confidence intervals (CI) 3.2% to 21.7%). The crude odds ratio (OR) and 95% CI was 0.50 (0.26 to 0.97). After adjusting for other factors, the trend persisted (OR 0.60, 95% CI 0.28 to 1.29). Having a previous Pap smear was the only significant predictor of intention to have screening (adjusted OR 2.54, 95% CI 1.03 to 6.21). Subgroup analysis showed no intervention effect in intended uptake between women at higher and lower risk of cervical cancer (p=0.59). CONCLUSIONS: Providing women with evidence based information on the risks, uncertainties, and the benefits of screening, is likely to deter some, but not differentially those at higher risk. PMID- 12883065 TI - Unemployment and suicide. Evidence for a causal association? AB - OBJECTIVES: To determine the independent associations of labour force status and socioeconomic position with death by suicide. DESIGN: Cohort study assembled by anonymous and probabilistic record linkage of census and mortality records. PARTICIPANTS: 2.04 million respondents to the New Zealand 1991 census aged 18-64 years. MAIN OUTCOME MEASURE: Suicide in the three years after census night. RESULTS: The age adjusted odds ratios (95% confidence intervals) of death by suicide among 25 to 64 year olds who were unemployed compared with employed were 2.46 (1.10 to 5.49) for women and 2.63 (1.87 to 3.70) for men. Similarly increased odds ratios were observed for the non-active labour force compared with the employed. Strong age only adjusted associations of suicide death with the socioeconomic factors of education (men only), car access, and household income were observed. Compared with those who were married on census night, the non married had odds ratios of suicide of 1.81 (1.22 to 2.69) for women and 2.08 (1.66 to 2.61) for men. In a multivariable model the association of socioeconomic factors with suicide reduced to the null. However, marital status and labour force status remained strong predictors of suicide death. Unemployment was also strongly associated with suicide death among 18-24 year old men. Sensitivity analyses suggested that confounding by mental illness might explain about half, but not all, of the association between unemployment and suicide. CONCLUSIONS: Being unemployed was associated with a twofold to threefold increased relative risk of death by suicide, compared with being employed. About half of this association might be attributable to confounding by mental illness. PMID- 12883066 TI - Women's life cycle and abortion decision in unintended pregnancies. AB - OBJECTIVE: To study the impact of sociodemographic, financial, and reproductive factors and of characteristics related to intimate relationships on the decisions of women in different age groups about whether or not to continue an unintended pregnancy. DESIGN: Cross sectional population based survey. SETTING: Telephone interview survey between September 2000 and January 2001 in France. From a representative sample (n=14704) of 18 to 44 year old women, those who in the past five years had an abortion or whose last pregnancy was unintended were oversampled (sampling fraction=100%, n=1034) while the other women were randomly selected (sampling fraction =19%, n=1829). Altogether, 2863 women answered the questionnaire. PARTICIPANTS: All women whose last pregnancy was unintended and ended in induced abortion or birth (n=645). MAIN RESULTS: Factors associated with the abortion decision varied strongly according to age. Younger women's abortion decisions were mainly related to being a student and being single. Wanting to stop childbearing when the desired number of children was achieved best explained the decision to have an abortion among 25 to 34 year old women. Older women chose abortion especially when childbearing did not fit their work situation or when the relationship with the partner was unstable. A high level of education of a woman and her partner increased the likelihood of abortion, especially among young women. CONCLUSIONS: The impact of socioeconomic and relationship factors on the decision to have an abortion is not the same at different stages in life, and refers to the social representations and perceptions of what good conditions are for being a mother. PMID- 12883067 TI - Preterm birth among African American and white women: a multilevel analysis of socioeconomic characteristics and cigarette smoking. AB - STUDY OBJECTIVE: Research shows that neighbourhood socioeconomic factors are associated with preterm delivery. This study examined whether cigarette smoking and individual socioeconomic factors modify the effects of neighbourhood factors on preterm delivery. DESIGN: Case-control study. SETTING: Moffit Hospital in San Francisco, California. PARTICIPANTS: 417 African American and 1244 white women, including all preterm and a random selection of term deliveries 1980-1990, excluding non-singleton pregnancies, congenital anomolies, induced deliveries, and women transported for special care. US census data from 1980 and 1990 were used to characterise the women's neighbourhoods, defined as census tracts. RESULTS: Cigarette smoking increased the risk of preterm delivery among both African American (OR=1.77, 95% confidence intervals (CI) (1.12 to 2.79)) and white women (OR=1.25, 95% CI (1.01 to 1.55)). However, cigarette smoking did not attenuate or modify the association of neighbourhood factors with preterm delivery. Among African American women, having public insurance modified the relation between neighbourhood unemployment and preterm delivery; among women without public insurance, the risk of preterm delivery was low in areas with low unemployment and high in areas with high unemployment, while among women with public insurance the risk of preterm delivery was highest at low levels of neighbourhood unemployment. CONCLUSIONS: Cigarette smoking was associated with preterm delivery, especially among African Americans. Adverse neighbourhood conditions had an influence on preterm delivery beyond that of cigarette smoking. The effects of some neighbourhood characteristics were different depending on individual socioeconomic status. Examining socioeconomic and behavioural/biological risk factors together may increase understanding of the complex causes of preterm delivery. PMID- 12883068 TI - Secular trends in sex ratios at birth in North America and Europe over the second half of the 20th century. AB - CONTEXT: A previous study showed that significantly more boys were born in southern latitudes in Europe than in northern latitudes and the converse pattern was observed in North America. OBJECTIVE: This study analyses secular trends in gender ratios for live births over the second half of the 20th century. DESIGN, SETTING, PARTICIPANTS: Analysis was carried out from a World Health Organisation dataset comprising live births over the above period. This included 127034732 North American and 157947117 European live births. MAIN OUTCOME MEASURES: Analysis of trends in gender ratios for countries in both continents. RESULTS: The findings show a highly significant overall decline in male births in both Europe and North America (p<0.0001), particularly in Mexico (p<0.0001). Interestingly, in Europe, male births declined in North European countries (latitude>40 degrees, p<0.0001) while rising in Mediterranean countries (latitude congruent with 35-40 degrees, p<0.0001). These trends produced an overall European male live birth deficit 238693 and a North American deficit of 954714 (total male live birth deficit 1193407). CONCLUSIONS: No reasonable explanation/s for the observed trends have been identified and the causes for these trends may well be multifactorial. PMID- 12883070 TI - Contextual risk factors for the common mental disorders in Britain: a multilevel investigation of the effects of place. AB - STUDY OBJECTIVE: To test hypotheses about associations between area level exposures and the prevalence of the most common mental disorders (CMD) in Britain. A statistically significant urban-rural gradient was predicted, but not a socioeconomic gradient, in the prevalence of CMD after adjusting for characteristics of individual respondents. The study tested the hypothesis that the effects of area level exposures would be greatest among those not in paid employment. DESIGN: Cross sectional survey, analysed using multilevel logistic and linear regression. CMD were assessed using the General Health Questionnaire (GHQ). Electoral wards were characterised using the Carstairs index, the Office of National Statistics (ONS) Classification of Wards, and population density. SETTING: England, Wales, and Scotland. PARTICIPANTS: Nearly 9000 adults aged 16 74 living in 4904 private households, nested in 642 electoral wards. MAIN RESULTS: Little evidence was found of statistically significant variance in the prevalence of CMD between wards, which ranged from 18.8% to 29.5% (variance 0.035, SE 0.026) (p=0.11). Associations between CMD and characteristics of wards, such as the Carstairs index, only reached statistical significance among those who were economically inactive (adjusted odds ratio for top v bottom Carstairs score quintile 1.58, 95% CI 1.08 to 2.31) (p<0.05). CONCLUSIONS: There may be multiple pathways linking socioeconomic inequalities and ill health. The effects of place of residence on mental health are greatest among those who are economically inactive and hence more likely to spend the time at home. PMID- 12883071 TI - Continuing inequality: gender and social class influences on self perceived health after a heart attack. AB - STUDY OBJECTIVE: To investigate the effect of social class and gender on self perceived health status for those recovering from an acute myocardial infarction. DESIGN: A longitudinal survey design was used, collecting both qualitative and quantitative data. Quantitative data are reported in this article, obtained by questionnaire over the first year after the event. SF-36 and EQ-5D (EuroQol) were used to measure self perceived health status. SETTING: Community based study in a city in the north of England. PARTICIPANTS: A consecutive sample of 229 people discharged from hospital after acute myocardial infarction. MAIN RESULTS: Overall gain in health status was found to be statistically significant over the year. Improvements were greatest in domains relating to role fulfillment and pursuit of normal and social activities. When analysed by gender, women showed poorer improvement than men, particularly in the domains relating to physical and social functioning. Analysed by social class, those without educational qualifications showed poorer improvement in pain experience and vitality. Access to a car was significant in avoiding physical limitations and promoting general health. CONCLUSIONS: Existing gradients between the health of women and men, and between the social classes, are maintained and probably exacerbated by the experience of acute illness, and health professionals need to be made aware of social groups who are at risk of poor rehabilitation. PMID- 12883072 TI - Mortality and temperature in Sofia and London. AB - STUDY OBJECTIVE: Heat and cold have been associated with increased mortality, independently of seasonal trends, but details are little known. This study explores associations between mortality and temperature in two European capitals Sofia and London-using four years of daily deaths, air pollution, and weather data. DESIGN: Generalised additive models were used to permit non-linear modelling of confounders such as season and humidity, and to show the shape of mortality-temperature relations-using both two day and two week average temperatures separately. Models with linear terms for heat and cold were used to estimate lags of effect, linear effects, and attributable fractions. PARTICIPANTS: 44701 all age all cause deaths in Sofia (1996-1999) and 256464 in London (1993-1996). MAIN RESULTS: In London, for each degree of extreme cold (below the 10th centile of the two week mean temperature), mortality increased by 4.2% (95% CI 3.4 to 5.1), and in Sofia by 1.8% (0.6 to 3.9). For each degree rise above the 95th centile of the two day mean, mortality increased by 1.9% (1.4 to 2.4) in London, and 3.5% (2.2 to 4.8) in Sofia. Cold effects appeared after lags of around three days and lasted-particularly in London-at least two weeks. Main heat effects occurred more promptly. There were inverse associations at later lags for heat and cold in Sofia. CONCLUSIONS: Average temperatures over short periods do not adequately model cold, and may be inadequate for heat if they ignore harvesting effects. Cold temperatures in London, particularly, seem to harm the general population and the effects are not concentrated among persons close to death. PMID- 12883073 TI - An adaptation of the Framingham coronary heart disease risk function to European Mediterranean areas. AB - AIM: To determine whether the Framingham function accurately predicts the 10 year risk of coronary disease and to adapt this predictive method to the characteristics of a Spanish population. METHOD AND RESULTS: A Framingham function for predicting 10 year coronary deaths and non-fatal myocardial infarction was applied to the population of the province of Gerona, Spain, where the cumulated incidence rate of myocardial infarction has been determined since 1988 by a specific registry. The prevalence of cardiovascular risk factors in this region of Spain was established in 1995 by a cross sectional study on a representative sample of 1748 people. The number of cases estimated by the Framingham function for 10 year coronary deaths and non-fatal myocardial infarction was compared with that observed. The Framingham function estimated 2425 coronary heart disease cases in women and 1181 were observed. In men, 9919 were estimated and 3706 were observed. Recalibrating the Framingham equations to the event rate and the prevalence of the risk factors in Gerona led to estimates very close to the number of cases observed in Gerona men and women. CONCLUSIONS: The Framingham function estimates more than doubled the actual risk of coronary disease observed in north east Spain. After calibration, the Framingham function became an effective method of estimating the risk in this region with low coronary heart disease incidence. PMID- 12883074 TI - Evidence for intersubunit interactions between S4 and S5 transmembrane segments of the Shaker potassium channel. AB - Voltage-gated potassium channels are transmembrane proteins made up of four subunits, each comprising six transmembrane (S1-S6) segments. S1-S4 form the voltage-sensing domain and S5-S6 the pore domain with its central pore. The sensor domain detects membrane depolarization and transmits the signal to the activation gates situated in the pore domain, thereby leading to channel opening. An understanding of the mechanism by which the sensor communicates the signal to the pore requires knowledge of the structure of the interface between the voltage sensing and pore domains. Toward this end, we have introduced single cysteine mutations into the extracellular end of S4 (positions 356 and 357) in conjunction with a cysteine in S5 (position 418) of the Shaker channel and expressed the mutants in Xenopus oocytes. We then examined the propensity of each pair of engineered cysteines to form a metal bridge or a disulfide bridge, respectively, by examining the effect of Cd2+ ions and copper phenanthroline on the K+ conductance of a whole oocyte. Both reagents reduced currents through the S357C,E418C double mutant channel, presumably by restricting the movements necessary for coupling the voltage-sensing function to pore opening. This inhibitory effect was seen in the closed state of the channel and with heteromers composed of S357C and E418C single mutant subunits; no effect was seen with homomers of any of the single mutant channels. These data indicate that the extracellular end of S4 lies in close proximity to the extracellular end of the S5 of the neighboring subunit in closed channels. PMID- 12883075 TI - Heterogeneity of toxicant response: sources of human variability. AB - While risk assessment models attempt to predict human risk to toxicant exposure, in many cases these models cannot account for the wide variety of human responses. This review addresses several primary sources of heterogeneity that may affect individual responses to drug or toxicant exposure. Consideration was given to genetic polymorphisms, age-related factors during development and senescence, gender differences associated with hormonal function, and preexisting diseases influenced by toxicant exposure. These selected examples demonstrate the need for additional steps in risk assessment that are needed to more accurately predict human responses to toxicants and drugs. PMID- 12883076 TI - Effect of 4-nonylphenol on cell proliferation and adipocyte formation in cultures of fully differentiated 3T3-L1 cells. AB - The effect of 4-nonylphenol (NP) on cell proliferation and adipocyte formation was examined in cultures of fully differentiated 3T3-L1 cells. Following the hormonal induction of differentiation into adipocytes, 3T3-L1 cells were treated for 8 days with or without NP. NP at 5 and 10 microg/ml increased the DNA content by 32% and 68%, respectively, compared with that of the untreated cultures, in which NP was absent during the treatment period. There were many more bromodeoxyuridine (BrdU)-positive cells in the NP-treated cultures, in which NP was present at a concentration of 10 microg/ml during the treatment period, compared to the untreated cultures. These results indicate that NP had the ability to stimulate the proliferation of fully differentiated 3T3-L1 cells. NP at 5 and 10 microg/ml decreased the triacylglycerol (TG) content by 26% and 58%, respectively, and decreased the lipoprotein lipase (LPL) activity by 51% and 71%, respectively. The lipid droplets in individual cells of the NP-treated cultures were smaller than those of the untreated cultures. The mRNA levels of LPL and adipocyte-specific fatty acid binding protein (aP2) were considerably lower in the NP-treated cultures than in the untreated cultures. Thus, NP also had the ability to inhibit adipocyte formation in cultures of fully differentiated 3T3-L1 cells. A study using an antiestrogen ICI 182,780 showed that the NP-stimulated cell proliferation was mediated partly by the estrogen receptor, while the NP induced inhibition of adipocyte formation was mediated by a mechanism other than the estrogen receptor. PMID- 12883077 TI - Aryl hydrocarbon receptor 2 mediates 2,3,7,8-tetrachlorodibenzo-p-dioxin developmental toxicity in zebrafish. AB - In order to use the zebrafish as a model vertebrate to investigate the developmental toxicity of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), it is essential to know whether one or both forms of the zebrafish aryl hydrocarbon receptor (AHR), zfAHR1 or zfAHR2, mediate toxicity. To determine the role of zfAHR2, an antisense morpholino approach was used to knock down translation of the protein. No effect of the zfahr2 morpholino (zfahr2-MO) was seen on normal development in embryos not treated with TCDD. Injection of embryos at the 1-2 cell stage with zfahr2-MO decreased TCDD-induced transcription of zfCYP1A mRNA until 96 h post fertilization (hpf), and immuno-histochemical detection of zfCYP1A protein in embryos at 72 hpf revealed a dramatic decrease in expression. The zfahr2-MO completely protected embryos from TCDD-induced edema and anemia and provided protection against TCDD-induced reductions in peripheral blood flow initially; however, a slight reduction in blood flow was observed at later times when the morpholino was no longer effective. Due to persistence of TCDD and decreasing effectiveness of the zfahr2-MO over time, the morpholino provided only transient protection against TCDD-induced inhibition of chondrogenesis of the lower jaw, and no protection against an effect of TCDD that was initiated late in development, blockade of swimbladder inflation. The zfahr2-MO did not protect embryos from TCDD-induced mortality but did produce a 48 h delay in its onset. Endpoints of TCDD developmental toxicity manifested in zfahr2 morphants at late stages of development, beyond 144 hpf, were clearly different from TCDD-exposed embryos injected with a control morpholino. Most strikingly, zfahr2 morphants exposed to TCDD never developed edema. Taken together, these results demonstrate that zfAHR2 mediates several endpoints of TCDD developmental toxicity in zebrafish. PMID- 12883078 TI - Cell proliferation and apoptosis are altered in mice deficient in the NF-kappaB p50 subunit after treatment with the peroxisome proliferator ciprofibrate. AB - We previously showed that the peroxisome proliferator ciprofibrate increases hepatic NF-kappaB DNA binding activity in rats, mice, and hepatoma cell lines. Here, we analyzed the response to ciprofibrate in mice that lack the NF-kappaB p50 gene (p50-/-). Wild-type and p50-/- mice were fed a diet with or without 0.01% ciprofibrate for 10 days. NF-kappaB DNA binding activity was present and increased after ciprofibrate treatment in wild-type mice, but was not detected in p50-/- mice. The untreated p50-/- mice had a higher level of hepatic cell proliferation, as measured by BrdU labeling, than did untreated wild-type mice. However, the increase in proliferation was greater in ciprofibrate-fed wild-type mice than in ciprofibrate-fed p50-/- mice. The apoptotic index was low in wild type mice in the presence or absence of ciprofibrate. Apoptosis was increased in untreated p50-/- mice compared to wild-type mice; apoptosis was reduced in p50-/- mice after ciprofibrate feeding. The c-Jun and JunB mRNA levels were higher in untreated p50-/- mice than in untreated control mice; c-Jun mRNA levels increased, whereas JunB mRNA levels decreased in both groups after ciprofibrate treatment. The c-Jun and JunB protein levels were the same in untreated wild-type and p50-/- mice and increased in both groups after ciprofibrate treatment. Several apoptosis-related mRNAs were higher in untreated p50-/- mice compared to untreated control mice; expression of these genes increased in both groups after ciprofibrate treatment. These data indicate that NF-kappaB contributes to the proliferative and apoptotic changes that occur in the liver in response to ciprofibrate. PMID- 12883079 TI - Modeling and predicting immunological effects of chemical stressors: characterization of a quantitative biomarker for immunological changes caused by atrazine and ethanol. AB - Previous studies demonstrate that the effects of one chemical stressor on selected immunological parameters can be predicted on the basis of the area under the corticosterone concentration vs. time curve. However, it is not clear if this is applicable to other chemical stressors. The present study was conducted to determine if the stress-induced immunological effects of atrazine and ethanol could be predicted, and if it is feasible to use one immunological parameter as a biomarker of stress to predict the quantity of changes expected in other immunological parameters. The area under the corticosterone concentration-versus time curve (AUC) was measured in mice treated with ethanol (EtOH, 4, 5, 6, or 7 g/kg by oral gavage) or atrazine (ATZ, 100, 200, or 300 mg/kg, ip). The effects of the same dosages of these chemicals on thymus and spleen cellularity, lymphocyte subpopulations in the thymus and spleen, expression of MHC class II protein on splenocytes, antibody responses to keyhole limpet hemocyanin, and natural killer-cell activity were determined. Models were derived describing the relationship between corticosterone AUC and immunological changes induced by these chemicals. The results for these chemical stressors were more similar to results obtained from mice subjected to restraint stress than from mice treated with exogenous corticosterone. Some effects were greater than predicted on the basis of the stress response alone, indicating other mechanisms of immunotoxicity. One of the parameters (MHC class II expression) was evaluated as a predictive biomarker for stress-related immunosuppression, and the results suggest it could be suitable for that purpose. PMID- 12883080 TI - 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) alters the regulation and posttranslational modification of p27kip1 in lipopolysaccharide-activated B cells. AB - 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) alters B-cell differentiation, as evidenced by a marked decrease in immunoglobulin M (IgM) secretion and in the number of antibody-forming cells (AFC) induced by antigenic stimulation. The objective of the present studies was to evaluate the effect of TCDD on the level of p27kip1, a cyclin-dependent kinase inhibitor that is a critical regulator of cellular differentiation. In the well-characterized B-cell line, CH12.LX, a modest decrease in p27kip1 was observed during the initial 24-h post-LPS (lipopolysaccharide) activation, which then gradually increased above background at 48 and 72 h. Conversely, in the presence of TCDD, p27kip1 was not induced and remained unchanged from LPS unstimulated cells throughout the entire 72-h period post-LPS activation. In addition, Western blotting revealed that TCDD treatment altered the profile of p27kip1 migration as compared to the LPS-activated control. Time-of-addition studies demonstrated that the greatest sensitivity of p27kip1 to TCDD treatment occurred within the initial 24-h post-LPS activation. Interestingly, LPS-induced Ig kappa light chain and IgM secretion also exhibited the greatest period of sensitivity (i.e., inhibition) to TCDD during the first 24 h post-LPS activation. In addition, TCDD markedly suppressed the LPS-induced differentiation of CH12.LX cells into IgM secreting AFC, with a modest but cumulative effect on cell proliferation over a 72-h period. Collectively, these findings show that TCDD altered the cellular concentration and posttranslational modification of p27kip1 in this activated B-cell line model, which occurred concomitantly with altered B-cell differentiation and suggests that cyclin dependent kinase inhibitors may be an important intracellular target in TCDD mediated inhibition of B-cell differentiation. PMID- 12883081 TI - Vinyl acetate decreases intracellular pH in rat nasal epithelial cells. AB - Vinyl acetate is a synthetic organic ester that has been shown to produce nasal tumors in rats following exposure to 600 ppm in air. The proposed mechanism of action involves the metabolism of vinyl acetate by carboxylesterases and the production of protons leading to cellular acidification. While vinyl acetate induced decreases in intracellular pH (pHi) have been demonstrated in rat hepatocytes, comparable data from nasal epithelial cells do not exist. Using an in vitro assay system, we have determined the effects of vinyl acetate exposure on pHi in respiratory and olfactory nasal epithelial cells from rats. The respiratory and olfactory epithelial cells were isolated from dissected maxillo- and ethmoturbinates by enzyme digestion. The cells were plated; loaded with the pH-sensitive dye, carboxyseminaphthorhodafluor-1 (SNARF-1); and observed using confocal microscopy. Individual cellular analysis demonstrated that both respiratory and olfactory epithelial cells responded to vinyl acetate exposures with a dose-dependent decrease in pHi. Changes occurred at 100 microM but reached a plateau above 250 microM. Maximal decreases in pHi were 0.3 pH unit in respiratory epithelial cells. While pHi values were normally distributed for the respiratory epithelial cells, the olfactory epithelial cells demonstrated a bimodal distribution, indicating at least two populations of cells, with only one population of cells responding to vinyl acetate. Acidification in these cells did not plateau but continued to increase at 1000 microM. Bis(p-nitrophenyl)phosphate (BNPP), a carboxylesterase inhibitor, was able to attenuate the vinyl acetate induced decrease in pHi. Data obtained from the isolated cells were validated using tissue explants. These results are consistent with the proposed mode of action for vinyl acetate and supply further data for developing appropriate risk assessments for vinyl acetate exposure. PMID- 12883082 TI - The future of regulatory toxicology: impact of the biotechnology revolution. AB - The molecular biology revolution and the advent of genomic and proteomic technologies are facilitating rapid advances in our understanding of the molecular details of cell and tissue function. These advances have the potential to transform toxicological and clinical practice, and are likely to lead to the supplementation or replacement of traditional biomarkers of cellular integrity, cell and tissue homeostasis, and morphological alterations that result from cell damage or death. New technologies that permit simultaneous monitoring of many hundreds, or thousands, of macro- and small molecules ("-omics" technologies) promise to allow functional monitoring of multiple (or perhaps all) key cellular pathways simultaneously. Elucidation of cellular responses to molecular damage, including evolutionarily conserved inducible molecular defense systems, suggests the possibility of new biomarkers based on molecular responses to functional perturbations and cellular damage. Our improved understanding of the molecular basis of various pathologies suggests that monitoring specific molecular responses may provide improved prediction of human outcomes. Responses that can be monitored directly in the human should provide "bridging biomarkers" that may eliminate much of the current uncertainty in extrapolating from laboratory models to human outcome. Another aspect of genomics is our enhanced ability to associate DNA sequence variations with biological outcomes and individual sensitivity. The human genome sequence has revealed that sequence variations are very common, and may be an important determinant of variation in biological outcomes. The impending availability of a complete human haplotype map linked to standard genetic markers greatly facilitates identification of genetic variations that convey sensitivity or resistance to chemical exposures. Genetic approaches have already linked a large number of genetic variants (polymorphisms) with human diseases and adverse reactions from exposure to drugs or toxicants, suggesting an important role in sensitivity to drugs and environmental agents, disease susceptibilities, and therapeutic responses. As these opportunities are transformed into reality, regulatory toxicological practice is likely to be shaped in the future by the combination of conventional pathology, toxicology, molecular genetics, biochemistry, cell biology, and computational bio-informatics resulting in the broad application of molecular approaches to monitoring functional disturbances. PMID- 12883083 TI - Use of a low-density microarray for studying gene expression patterns induced by hepatotoxicants on primary cultures of rat hepatocytes. AB - In the field of gene expression analysis, DNA microarray technology is having a major impact on many different areas including toxicology. For instance, a number of studies have shown that transcription profiling can generate the information needed to assign a compound to a mode-of-action class. In this study, we investigated whether compounds inducing similar toxicological endpoints produce similar changes in gene expression. In vitro primary rat hepatocytes were exposed to 11 different hepatotoxicants: acetaminophen, amiodarone, clofibrate, erythromycin estolate, isoniazid, alpha-naphtylylisothiocyanate, beta naphtoflavone, 4-pentenoic acid, phenobarbital, tetracycline, and zileuton. These molecules were selected on the basis of their variety of hepatocellular effects observed such as necrosis, cholestasis, steatosis, and induction of CYP P450 enzymes. We used a low-density DNA microarray containing 59 genes chosen as relevant toxic and metabolic markers. The in vitro gene expression data generated in this study were generally in good agreement with the literature, which mainly concerns in vivo data. Furthermore, gene expression profiles observed in this study have been confirmed for several genes by real-time PCR assays. All the tested drugs generated a specific gene expression profile. Our results show that even with a relatively limited gene set, gene expression profiling allows a certain degree of classification of compounds with similar hepatocellular toxicities such as cholestasis, necrosis. The clustering analysis revealed that the compounds known to cause steatosis were linked, suggesting that they functionally regulate similar genes and possibly act through the same mechanisms of action. On the other hand, the drugs inducing necrosis and cholestasis were pooled in the same cluster. The drugs arbitrarily classified as the CYP450 inducers formed individual clusters. In conclusion, this study suggests that low density microarrays could be useful in toxicological studies. PMID- 12883084 TI - Effects of inorganic HgCl2 on adipogenesis. AB - Mercury is a common pollutant that alters glucose metabolism in adipocytes; however, the effect of HgCl2 on differentiating adipocytes and their subsequent metabolic function is not well understood. Two adipocyte models, the 3T3-L1 and C3H10T1/2 (10T1/2) cell lines, were differentiated in the presence of HgCl2. To assess the amount of differentiation in a population, markers of differentiation (i.e., PPARgamma and GLUT 4 expression and lipid accumulation) and functions of adipocytes (i.e., glucose transport and insulin-induced glucose transport) were measured. HgCl2 exposure significantly decreased the number of phenotypic adipocytes and PPARgamma expression in both 3T3-L1 and 10T1/2 cells without effects on cell viability. GLUT 4 was significantly reduced by HgCl2 treatment in 10T1/2 but not 3T3-L1 cells. Exposure to HgCl2 during differentiation increased basal glucose uptake in a dose-dependent manner (up to 2.5-fold) and decreased insulin-induced glucose uptake in 3T3-L1 adipocytes. In contrast, HgCl2 had little effect on basal or insulin-induced glucose uptake in 10T1/2 cells, possibly due to their lower insulin responsiveness. We examined the effect of HgCl2 exposure on signaling event involved in differentiation of adipocytes and cellular stress, namely, the phosphorylation of ERK1/2 and JNK, respectively. HgCl2 exposure had no effect on ERK1/2 phosphorylation in either cell line, increased JNK phosphorylation in the 10T1/2, and had no effect on JNK phosphorylation in 3T3-L1 cells. These data indicate HgCl2 exposure can inhibit the differentiation of fibroblasts into adipocytes as well as influence signaling events and the subsequent metabolic activity of differentiated adipocytes. PMID- 12883085 TI - Effect of dietary co-administration of sodium selenite on sodium arsenite-induced ovarian and uterine disorders in mature albino rats. AB - The subchronic treatment of mature female Wistar-strain albino rats in diestrous phase with sodium arsenite at a dose of 0.4 ppm/100 g body weight/rat/day via drinking water for period of 28 days (seven estrous cycles) caused a significant reduction in the plasma levels of leutinizing hormone (LH), follicle-stimulating hormone (FSH), and estradiol along with a significant decrease in ovarian activities of delta five, 3 beta-hydroxysteroid dehydrogenase (Delta5,3beta-HSD), and 17 beta-hydroxysteroid dehydrogenase (17beta-HSD) followed by a reduction in ovarian and uterine peroxidase activities. A significant weight loss of the ovary and uterus was also observed after this treatment, along with a prolonged diestrous phase and a high accumulation of arsenic in the plasma and these organs. Moreover, sodium arsenite was also responsible for ovarian follicular and uterine cell degeneration characterized by a high number of regressing follicles and a reduction in the uterine luminal diameter, respectively, in comparison with the controls. A dietary supplementation of sodium selenite at the dose of 0.6 mg/100 g body weight/rat/day for a period of 28 days along with arsenic treatment minimized the gonadal weight loss significantly and increased the activities of the ovarian steroidogenic enzymes as well as the ovarian and uterine peroxidase at the control level. Selenium was also able to increase the plasma levels of LH, FSH, and estradiol toward the control level. Vaginal smears showed normal estrous cyclicity in sodium selenite-supplemented arsenic-treated rats along with lower arsenic levels in the plasma and gonadal tissue in comparison with arsenic-only treated rats. Histological sections of ovary and uterine tissues in the control and experimental groups confirmed that sodium selenite supplementation was able to prevent arsenic-induced histopathological changes in the ovary and uterus. Plasma levels of norepinephrine and dopamine in the midbrain and diencephalon decreased significantly, whereas the serotonin level was increased significantly after 28 days of sodium arsenite treatment. All of these parameters were, in most cases, unchanged from the control level when sodium selenite was co-administered with sodium arsenite. Arsenic intoxication was also associated with increased liver weight and elevation in the activities of hepatic and renal acid phosphatase, alkaline phosphatase, and transaminases, but selenium co administration was not able to change these toxic effects of arsenic. The results of our experiments indicate the significant protective action of sodium selenite on arsenic-induced toxicity in the female reproductive system, while there was no significant protective effect of selenium on arsenic-induced toxicity in other organs. PMID- 12883086 TI - Developmental toxicity and genotoxicity studies of 1,1,1,3,3,3-hexachloropropane (HCC-230fa) in rats. AB - The potential developmental toxicity and the in vitro and in vivo genotoxicity of HCC-230fa were assessed. In the developmental toxicity study, groups of 25 mated Crl:CD(R)(SD)BR rats were exposed (whole body) by inhalation to HCC-230fa over days 7-21 of gestation; the day of confirmed mating was designated as gestation day 1 (GD1). Exposures were 6 h per day at concentrations of 0, 0.5, 2.5, or 25 ppm. Body weight, food consumption, and clinical observation data were collected during the study. On day 22 of gestation, the dams were euthanized and examined grossly. The fetuses were removed and subsequently weighed, sexed, and examined for external, visceral, head, and skeletal alterations. Evidence of maternal and developmental toxicity was observed at 25 ppm and was noted as significant, compound-related reductions in mean maternal body weight, weight change, and food consumption. Significant fetal effects also were observed at 25 ppm as compound related reductions in mean fetal weight and increased fetal malformations (filamentous tail, situs inversus, absent vertebrae) and variations (rudimentary cervical ribs, delayed sternebral ossification). There was no evidence of either maternal or developmental toxicity at 0.5 or 2.5 ppm. The genotoxicity of HCC 230fa was examined in a bacterial reversion assay and in erythrocyte micronucleus studies in two species by different routes of administration. No increases in the number of revertants were observed in the bacterial reversion assay. In one micronucleus study, HCC-230fa was administered by inhalation to rats as part of a 90-day study at doses indicated above. For the second study, ICR mice were given a single ip dose at 0, 166, 330, or 660 mg/kg. In both micronucleus studies, a significant increase in micronucleated erythrocytes was observed. The results of these studies suggest that HCC-230fa affects rapidly dividing cells and may have long-term consequences for occupational exposures. PMID- 12883087 TI - Dieldrin stimulates biliary excretion of 14C-benzo[a]pyrene polar metabolites but does not change the biliary metabolite profile in rainbow trout (Oncorhyncus mykiss). AB - Activities of hepatic microsomal and cytosolic epoxide hydrolases, accumulation of dieldrin in liver, and in vivo metabolism and disposition of the polycyclic aromatic hydrocarbon (PAH), benzo[a]pyrene (BP), were examined in rainbow trout pretreated with dieldrin, a chlorinated cyclodiene insecticide. Rainbow trout were fed 0.3 mg dieldrin/kg/day for 9 weeks and the same dose of dieldrin for 9 weeks, followed by 3 weeks on control diet (12 weeks). Fish then received an intraperitoneal (ip) challenge dose of 14C-BP (10 micromol/kg). Dieldrin pretreatment significantly elevated the concentration of 14C-BP in bile (142% and 200% at 9 and 12 weeks, respectively), but not liver or fat. Extraction of bile subsamples confirmed dieldrin pretreatment significantly stimulated total biliary excretion of 14C-BP polar metabolites (244% and 221% at week 9 and 12, respectively). The complex metabolism of BP characterized the in vivo state of the CYP system, UDP-glucuronyltransferases, and sulfotransferases. Bile was extracted and then hydrolyzed by beta-glucuronidase and arylsulfatase to regenerate BP metabolites conjugated by phase II enzymes. Evaluation of biliary polar metabolite profiles of 14C-BP revealed no significant differences between control and dieldrin-fed fish. There was no selective enhancement of any particular metabolite, or formation of a novel metabolite with dieldrin pretreatment. This research confirmed that enhanced biliary excretion, following chronic dieldrin exposure, was not explained by induction of xenobiotic metabolizing enzymes. The results are consistent with induction of hepatic intracellular trafficking proteins in dieldrin-fed fish. PMID- 12883088 TI - Acrylamide: a comparison of metabolism and hemoglobin adducts in rodents following dermal, intraperitoneal, oral, or inhalation exposure. AB - Acrylamide (AM), which is used to manufacture polymers, is carcinogenic and a reproductive and neurological toxicant. The objective of this study was to compare the metabolism of AM administered orally (po), dermally, intraperitoneally (ip), or by inhalation, and to measure the hemoglobin adducts produced. Rats and mice were exposed to 2.9 ppm [1,2,3-13C] and [2,3-14C]AM for 6 h. [2,3-14C]AM (162 mg/kg) or [1,2,3-13C]AM (13 8 mg/kg) in water was administered dermally to rats for 24 h, and [1,2,3-13C]AM was administered ip (47 mg/kg). Urine and feces were collected for 24 h. Urine was the major elimination route in rats (ip, 62% and po, 53% of the dose; dermal, 44% of the absorbed dose; inhalation, 31% of the recovered radioactivity) and mice (inhalation, 27% of the recovered radioactivity). Signals in the 13C-NMR spectra of urine were assigned to previously identified metabolites derived from AM glutathione conjugation (AM GSH) and conversion to glycidamide (GA). AM-GSH was a major metabolic route in rats accounting for 69% (ip), 71% (po), 52% (dermal), and 64% (inhalation). In mice, AM-GSH accounted for only 27% (inhalation) of the total urinary metabolites. The remaining urinary metabolites were derived from GA. Valine hemoglobin adducts of AM and GA were characterized using liquid chromatography mass spectrometry. The ratio of AM to GA adducts paralleled the flux through pathways based on urinary metabolites. This study demonstrates marked species differences in the metabolism and internal dose (Hb-adducts) of AM following inhalation exposure and marked differences in uptake comparing dermal with po and ip administration. PMID- 12883089 TI - Progressive alterations in global and GC-rich DNA methylation during tumorigenesis. AB - DNA methylation plays a key role in the regulation of gene expression, and failure to maintain normal patterns of methylation often contributes to carcinogenesis. We have characterized progressive methylation changes during the promotion stage of carcinogenesis using a SENCAR mouse skin initiation/promotion tumorigenesis model. Mice were initiated with a dermal application of 75 microg dimethylbenz[a]anthracene (DMBA) and promoted with 9, 18, 27, and 36 mg cigarette smoke condensate (CSC) thrice weekly for time periods up to 29 weeks, when a large increase in tumor number was produced by the highest three doses. Global and GC-specific methylation were assessed using SssI methylase and arbitrarily primed PCR, respectively. Changes in GC-specific methylation were dose- and time dependent. CSC doses required to detect these changes were 27 mg at 6 weeks and 18 mg at 9 weeks. This effect appears to be reversible; changes in GC-specific methylation were less marked after 9 weeks promotion with 27 mg CSC followed by 6 weeks of recovery in comparison to 9 and 15 weeks promotion with 27 mg CSC and no recovery period. Both tumor and non-tumor tissue promoted with 27 mg CSC for 29 weeks exhibited changes in GC-specific methylation that were more pronounced in tumors. Tumor tissue was globally hypomethylated, whereas non-tumor tissue did not exhibit changes in global methylation. In conclusion, as expected for a mechanism underlying tumor promotion, CSC alters methylation in a threshold exhibiting, reversible, progressive fashion during promotion. Progressive alterations in global and GC-rich methylation appear to be mechanistically important during tumor promotion. PMID- 12883090 TI - Effects of postnatal exposure to mixtures of non-ortho-PCBs, PCDDs, and PCDFs in prepubertal female rats. AB - There are concerns that postnatal exposure to organochlorines present in breast milk could lead to adverse health effects. We reconstituted four mixtures of aryl hydrocarbon receptor (AhR) agonists (3 non-ortho polychlorinated biphenyls [PCBs], 6 polychlorinated dibenzodioxins [PCDDs], 7 polychlorinated dibenzofurans [PCDFs], or all 16 chemicals together [referred to as AhRM]) based on their concentrations in breast milk, and examined their effects following exposure by gavage from day 1 until day 20 of age. Female neonates received dosages of AhRM equivalent to 1, 10, 100, or 1000 times the amount consumed by an infant over the first 24 days of life. Other groups received the PCBs, the PCDDs, or the PCDFs at the 1000x level. All rats were sacrificed at 21 days of age. Changes in ethoxyresorufin-o-deethylase hepatic activity, thymus and body weights, and serum thyroxin were linked to the 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) toxic equivalents (TEQ) of the four mixtures (1000x-AhRM > PCDDs > PCBs > PCDFs). To test for AhRM antiestrogenicity, two additional groups received 1.5 microg/kg of 17alpha-ethynyl estradiol (EE) with or without the 1000x-AhRM. The AhRM had no effect on uterine weight or EE-stimulated uterine growth. The actions of the combined EE and AhRM treatments suggest additive effects in decreasing pentoxyresorufin-o-deethylase activity and spleen weight, but nonadditive/antagonistic effects on adrenal weight and serum thyroxin. In conclusion, (1) 10x-AhRM had no detectable effects, (2) TEQ values relate to observed toxicities, even when testing complex mixtures of AhR agonists, and (3) indications of tissue-specific additive and nonadditive/antagonistic effects, but no synergism, were observed when doses of AhRM were increased, or combined with EE. PMID- 12883091 TI - Application of the CFU-GM assay to predict acute drug-induced neutropenia: an international blind trial to validate a prediction model for the maximum tolerated dose (MTD) of myelosuppressive xenobiotics. AB - In a previous study of prevalidation, a standard operating procedure (SOP) for two independent in vitro tests (human and mouse) had been developed, to evaluate the potential hematotoxicity of xenobiotics from their direct and the adverse effects on granulocyte-macrophages (CFU-GM). A predictive model to calculate the human maximum tolerated dose (MTD) was set up, by adjusting a mouse-derived MTD for the differential interspecies sensitivity. In this paper, we describe an international blind trial designed to apply this model to the clinical neutropenia, by testing 20 drugs, including 14 antineoplastics (Cytosar-U, 5 Fluorouracil, Myleran, Thioguanine, Fludarabine, Bleomycin, Methotrexate, Gemcitabine, Carmustine, Etoposide, Teniposide, Cytoxan, Taxol, Adriamycin); two antivirals (Retrovir, Zovirax,); three drugs for other therapeutic indications (Cyclosporin, Thorazine, Indocin); and one pesticide (Lindane). The results confirmed that the SOP developed generates reproducible IC90 values with both human and murine GM-CFU. For 10 drugs (Adriamycin, Bleomycin, Etoposide, Fludarabine, 5-Fluorouracil, Myleran, Taxol, Teniposide, Thioguanine, and Thorazine), IC90 values were found within the range of the actual drug doses tested (defined as the actual IC90). For the other 10 drugs (Carmustine, Cyclosporin, Cytosar-U, Cytoxan, Gemcitabine, Indocin, Lindane, Methotrexate, Retrovir, and Zovirax) extrapolation on the regression curve out of the range of the actual doses tested was required to derive IC90 values (extrapolated IC90). The model correctly predicted the human MTD for 10 drugs out of 10 that had "actual IC90 values" and 7 drugs out of 10 for those having only an extrapolated IC90. Two of the incorrect predictions (Gemcitabine and Zovirax) were within 6 fold of the correct MTD, instead of the 4-fold range required by the model, whereas the prediction with Cytosar-U was approximately 10-fold in error. A possible explanation for the failure in the prediction of these three drugs, which are pyrimidine analogs, is discussed. We concluded that our model correctly predicted the human MTD for 20 drugs out of 23, since the other three drugs (Topotecan, PZA, and Flavopiridol) were tested in the prevalidation study. The high percentage of predicitivity (87%), as well as the reproducibility of the SOP testing, confirm that the model can be considered scientifically validated in this study, suggesting promising applications to other areas of research in developing validated hematotoxicological in vitro methods. PMID- 12883092 TI - Effect of N-acetylcysteine on acetaminophen toxicity in mice: relationship to reactive nitrogen and cytokine formation. AB - The relationship between acetaminophen (APAP) reactive metabolite formation, nitrotyrosine (NT) production, and cytokine elevation in APAP toxicity was investigated. Mice were dosed with 300 mg/kg of APAP and sacrificed at 1, 2, 4, 8, and 12 h. Serum aspartate aminotransferase (AST) was elevated by 4 h. The relative amount of NT correlated with toxicity and was localized in the necrotic cells. IL-1b was increased at 1 h, whereas IL-6, MIP-2, and MCP-1 were increased by 4-8 h. To determine the importance of reversible versus toxic events, N acetylcysteine (NAC) was administered to mice either before APAP or 1, 2, or 4 h after APAP. The animals were sacrificed at 12 h. NAC treatment before APAP resulted in serum AST, serum nitrate plus nitrite as a measure of nitric oxide (NO) production, and hepatic cytokine levels that were similar to the controls. No APAP protein adducts or NT was present in these animals. In mice treated with NAC at 1 h, cytokines and serum AST were normal at 12 h, but APAP protein adducts were present in the hepatic centrilobular areas. No NT was present in these animals. In mice treated with NAC at 2 h and sacrificed at 12 h, serum AST was reduced by 80%. APAP adducts and NT were present in the centrilobular areas. Mice receiving NAC at 4 h had no protection from toxicity and serum nitrate plus nitrite. The NT and cytokine levels were similar to those of mice receiving APAP alone. The data suggest a relationship between metabolic events in APAP toxicity and the upregulation of NO, and IL-1b. IL-6, MIP-2, and MCP-1 appear to follow the toxicity. While it is a pre-requisite event, covalent binding per se does not appear to be a toxic event in the development of toxicity. PMID- 12883093 TI - In utero through lactational exposure to ethinyl estradiol induces cleft phallus and delayed ovarian dysfunction in the offspring. AB - Most of the attention currently focused on endocrine-active chemicals is directed to their effects on the development of offspring exposed to them in utero or during the neonatal period. Pregnant Crj:CD(SD)IGS rats were given ethinyl estradiol (EE) orally in doses of 0.5-50 microg/kg/day from gestational day 7 to postnatal day 18, and their offspring were examined for its effects. Our previous study according to a similar protocol demonstrated the occurrence of cleft phallus in the female offspring exposed to 50 microg/kg of EE in utero and during the lactation period. The present study was designed to assess (1) the reproducibility of the induction of cleft phallus, (2) the fertility of female rats with cleft phallus, and (3) whether any delayed effects, possibly delayed anovulation, were induced. At 50 microg/kg cleft phallus was observed in almost all of the female offspring, and slight retardation of body weight gain was detected in both sexes. At 15-17 weeks of age the animals with cleft phallus could copulate and had fertility comparable to the control group. At 6 months of age, on the other hand, 6/8 of the female offspring at 50 microg/kg exhibited abnormal cyclicity, including persistent estrus, and histological examination revealed follicular cysts and absence of corpora lutea in the ovaries of the rats with persistent estrus. These findings are consistent with delayed anovulation syndrome. The results suggest that observation of cyclicity at 6 months old is able to detect possible delayed ovarian dysfunction induced by perinatal exposure to chemicals. PMID- 12883095 TI - Major depression, physical illness, and suicidal ideation in primary care. AB - OBJECTIVE: To determine the association between major depression and suicidal ideation and the role of physical illness in this link among primary care patients. METHOD: More than 3,000 randomly selected primary care patients at eight sites across the United States completed the PRIME-MD PHQ, a screen for mental disorders for use in primary care. Physicians independently diagnosed physical illnesses. Multiple logistic regression analyses were used to determine the relationship between PRIME-MD depression, physical illness, and suicidal ideation. RESULTS: Pulmonary disease was associated with an increased likelihood of suicidal ideation, even among patients without major depression [odds ratio = 1.9 (1.04, 3.4)]. There was evidence of statistical interaction between pulmonary disease and depression in increasing the odds of suicidal ideation. Specifically, patients with pulmonary disease without depression, those with depression without pulmonary disease, and patients with both pulmonary disease and depression had significantly increased odds of suicidal ideation with odd ratios of 1.9 (1.04, 3.4), 7.4 (5.6, 9.7), and 9.6 (5.1, 18.0), respectively. CONCLUSIONS: These data suggest that some physical disorders may be associated with increased suicidal ideation in primary care and may also play a role in the relationship between depression and suicidal ideation among primary care patients. Primary care physicians may wish to engage in an in-depth evaluation of psychiatric problems, especially current suicidal ideation, among patients with specific ongoing physical illnesses. PMID- 12883096 TI - Depression and medical comorbidity: red flags for current suicidal ideation in primary care. PMID- 12883097 TI - Depressive symptoms and mortality two years after coronary artery bypass graft surgery (CABG) in men. AB - OBJECTIVES: Depression has been related to mortality in patients with CAD and to medical morbidity after CABG; however, prior studies have not examined the contribution of presurgical depressive symptoms to mortality after CABG. The purpose of this study was to determine the independent contribution of presurgical symptoms of depression to 2-year cardiac mortality after CABG. METHODS: Eighty-nine consecutive veteran nonemergent CABG patients recruited between December 1996 and June 1998 completed the BDI 1 to 7 days before surgery. Mortality risk was assessed by medical co-morbidity and RIS. RESULTS: Significant univariate contributions to two-year cardiovascular mortality were found for RIS (chi2 = 6.57, p <.01), history of CHF (chi2 = 4.94, p <.02), history of COPD (chi2 = 5.19, p <.02), and elevated depressive symptoms (chi2 = 4.70, p <.03). The multivariate model revealed that the RIS (chi2 = 4.70, p <.03) and elevated depressive symptoms (chi2 = 3.86, p <.05) remained significant in the prediction of 2-year cardiovascular mortality, with no other variables being found significant. CONCLUSIONS: Elevated depressive symptoms before CABG surgery appear to be an important independent contributor to long-term mortality. Future research should focus on replication with larger, more diverse samples, and identification of pathophysiological mechanisms. PMID- 12883098 TI - An international perspective on gender differences in anxiety early after acute myocardial infarction. AB - OBJECTIVE: Higher anxiety is linked to poorer outcomes after acute myocardial infarction (AMI), including increased in-hospital reinfarction and potentially life-threatening complications. If clinicians can identify patients at greatest risk for anxiety after AMI, they can institute early treatment. Previous research on the influence of gender on the incidence of anxiety post-AMI reflects inconsistent findings, and differences across cultures have not been studied. Therefore, the purposes of this study were to determine: 1) whether there are gender differences in anxiety in a diverse international sample of AMI patients, and 2) whether there was an interaction between gender and sociodemographic and clinical variables thought to influence anxiety. METHODS: In this prospective, comparative study, 912 AMI patients were enrolled from Australia, South Korea, Japan, England, and the United States. Anxiety was assessed, using the Brief Symptom Inventory, within the first 72 hours of admission to the hospital for AMI symptoms. RESULTS: Women had higher anxiety levels than men (0.76 +/- 0.90 vs. 0.57 +/- 0.70, p =.005), and this pattern of higher anxiety in women was seen in each country studied. Neither sociodemographic nor clinical variables interacted with gender to influence anxiety. CONCLUSION: Across a variety of cultures, women have higher anxiety than men after AMI and this relationship is independent of age, education level, marital status, or presence of comorbidities or severity of AMI. PMID- 12883099 TI - High plasma levels of N-terminal pro-atrial natriuretic peptide associated with low anxiety in severe heart failure. AB - OBJECTIVE: Plasma levels of natriuretic peptides are elevated in congestive heart failure (CHF). These peptides show anxiolytic properties in studies of rodents and patients with panic disorder, but their possible effect on anxiety has never been studied in cardiac patients. We therefore assessed associations of the Atrial Natriuretic Pro-Peptide (pro-ANP) with anxiety in patients with CHF and controls. METHOD: This was a cross-sectional study of 119 patients (46 with CHF, 76 controls with cardiovascular risk factors) in a tertiary care center. The study included assessment of CHF severity, ejection fraction, pro-ANP (microtiter assay), and psychosocial status (self-rating questionnaires for anxiety, depression, vital exhaustion, and quality of life). RESULTS: The diagnosis and severity of CHF was significantly related to pro-ANP levels, bad physical quality of life, vital exhaustion, and depression. However, there was no significant effect of disease severity on anxiety. In CHF patients, pro-ANP was negatively correlated with anxiety (rho = -0.30, p =.041). In the whole group, anxiety was independently predicted by vital exhaustion, depression, and younger age (overall adjusted R2 = 0.48). Pro-ANP plasma levels showed an additional, inverse association with anxiety (beta = -0.17, p =.013, adjusted R2 = 0.50). Predicted mean anxiety scores derived from this model showed a good fit with anxiety scores observed in subgroups defined by CHF severity. CONCLUSION: Pro-ANP plasma levels are independently and inversely related to anxiety. Even in severe CHF with severely compromised quality of life, anxiety tends to decrease with high pro-ANP levels. This might be part of a negative feedback loop limiting psychological distress and its adverse autonomic consequences in severe heart failure. PMID- 12883100 TI - Joint effect of hostility and severity of depressive symptoms on plasma interleukin-6 concentration. AB - OBJECTIVE: Although interleukin (IL)-6 plays a significant role in cardiovascular disease, little is known about its relation to psychological risk factors, such as hostility and severity of depressive symptoms. The current study examined the joint effects of severity of depressive symptoms and hostility on plasma IL-6 in a sample of 90 healthy, nonsmoking men. METHODS: After an overnight fast, blood samples for plasma IL-6 and fasting lipids were collected on the same day that the Beck Depression Inventory (BDI) and the Cook-Medley hostility (Ho) scale were administered. Plasma IL-6 was determined using enzymatic-linked immunosorbent assay (ELISA). RESULTS: Analyses of logarithmically normalized plasma IL-6 adjusting for age, body mass index (BMI), fasting total cholesterol, high density lipoprotein (HDL), and resting diastolic blood pressure (DBP) revealed a significant BDI by Ho interaction (p =.026). Post hoc decomposition revealed that Ho was correlated with log-normalized plasma IL-6 (r = 0.59, p =.025) but only among men with BDI scores of 10 and above. Alternatively, BDI was correlated with log-normalized plasma IL-6 (r = 0.61, p =.003) but only among men with Ho scores of 23 and higher. Comparisons among BDI/Ho groups indicated that men with high scores on both the BDI and the Ho exhibited the highest median levels of plasma IL-6. CONCLUSION: Hostile men who exhibited above normal levels of depressive symptoms had higher plasma levels of IL-6 suggestive of a subpopulation at increased risk for future cardiac events. PMID- 12883101 TI - Medically unexplained physical symptoms, anxiety, and depression: a meta-analytic review. AB - OBJECTIVE: Our objective was to review and compare, with meta-analytic methods, observational studies on the association of medically unexplained physical symptoms, anxiety, and depression with special emphasis on healthy and organically ill control groups and on different types of symptoms, measures, and illness behavior. METHODS: A search of MEDLINE and PsycLIT/PsycINFO for abstracts from 1980 to April 2001 was performed; principal investigators in the field were contacted and article reference lists were used to retrieve additional relevant articles. Two hundred forty-four studies were included on the basis of consensus ratings if they fulfilled seven of eight inclusion criteria pertaining to diagnostic accuracy and statistical appropriateness. Five hundred twenty-two studies were deferred or excluded. We focused specifically on the four functional somatic syndromes for which there were sufficient numbers for meta-analytic integration: irritable bowel syndrome (IBS), nonulcer dyspepsia (NUD), fibromyalgia (FM), and chronic fatigue syndrome (CFS). Data were extracted independently by two authors according to a prespecified coding manual with up to 70 parameters per study. RESULTS: Effect sizes for the association of the four functional somatic syndromes with depression and anxiety were of moderate magnitude but were highly significant statistically when compared with healthy persons and controls with medical disorders of known organic pathology. Moreover, this association was significant whether depression was measured with or without somatic items. Chronic fatigue syndrome is characterized by higher scores of depression, fibromyalgia by lower scores of anxiety than irritable bowel syndrome. Consulting behavior and severity of somatization is related to higher levels of anxiety and depression. CONCLUSIONS: Meta-analytic integration confirms that the four functional somatic syndromes (IBS, NUD, FM, CFS) are related to (but not fully dependent on) depression and anxiety. At present, there is only limited meta-analytic evidence for the same sort of association for medically unexplained physical symptoms in general. In view of the relative independence from depression and anxiety, classification and treatment of these symptoms and syndromes as "common mental disorders" does not seem fully appropriate. PMID- 12883102 TI - Medical inpatients at risk of extended hospital stay and poor discharge health status: detection with COMPRI and INTERMED. AB - OBJECTIVE: To detect the patients in medical wards at risk of extended LOS and poor discharge health status with the use of complexity prediction instrument (COMPRI) and interdisciplinary medicine (INTERMED) instruments. METHODS: STUDY 1: In a sample of 275 consecutively admitted medical inpatients, a hierarchical cluster analysis on INTERMED variables was performed. The clusters were compared on length of hospital stay (LOS) and Short Form 36 (SF-36) at discharge. STUDY 2: Receiver operating characteristic (ROC) analysis was used to optimal cut-off points for the COMPRI and INTERMED. Patients detected with COMPRI and INTERMED were then compared with undetected patients on LOS and SF-36. RESULTS: STUDY 1: In concordance with previous findings, a cluster of patients with high biopsychosocial vulnerability was identified with significantly higher scores on LOS (p <.05) and lower scores on SF-36 (p <.001) than patients in other clusters. STUDY 2: A cut-off point for the COMPRI of 5/6 was found to detect patients at risk of long LOS. A cut off score for the INTERMED of 20/21 was found to detect patients at risk of poor discharge health status. Patients detected with COMPRI and INTERMED had a significantly longer LOS (p <.001) and a poorer discharge health status (SF-36 MCS: p <.001; SF-36 PCS: p =.05) than nondetected patients. Of the detected patients, 37% had an extended hospital stay and poor discharge health status; of the nondetected patients, this was only 7%. CONCLUSIONS: The COMPRI-INTERMED can help to detect complex patients admitted to medical wards within the first days of admission, and rule out those with a small chance of poor outcomes. PMID- 12883103 TI - Effects of treatment for depression on fatigue in multiple sclerosis. AB - OBJECTIVE: There has long been a belief that depression contributes to fatigue in multiple sclerosis (MS) although supporting data are minimal at best. Clinical guidelines for the treatment of fatigue include recommendations for the treatment of depression in the absence of clear empirical support. The goal of this study was to examine the effects of treatment for depression on fatigue in MS. METHODS: Sixty patients with a relapsing form of MS and moderate to severe depression were randomly assigned to one of three validated 16-week treatments for depression: individual cognitive behavioral therapy, group psychotherapy, or sertraline. Assessments at baseline and treatment cessation included the primary outcome measure, fatigue assessment instrument (FAI), and Beck depression inventory (BDI). RESULTS: The total FAI and the global fatigue severity subscale were significantly reduced over the course of treatment (p values <.02). Other subscales did not change significantly. Secondary analyses showed change in global fatigue severity was associated with change in BDI (p =.03) but change in total FAI was only marginally related to change in BDI (p =.05). These relationships were due entirely to change in mood (p values <.02) and not to change in cognitive or vegetative symptoms (p values >.17). CONCLUSIONS: These findings suggest that treatment for depression is associated with reductions in the severity of fatigue symptoms, and that this relationship is due primarily to treatment related changes in mood. PMID- 12883104 TI - A twin study of depression symptoms, hypertension, and heart disease in middle aged men. AB - OBJECTIVE: Epidemiological and clinical studies have established an association between major depression and cardiovascular disease. We utilized a twin design to test whether there are common genetic and environmental risk factors underlying depression symptoms, hypertension and heart disease. METHODS: Association studies were conducted with 6,903 male-male twins from the Vietnam Era Twin Registry who responded to both a 1990 health questionnaire and a 1992 telephone administration of a structured psychiatric interview. Data from 2,731 complete twin pairs were used to fit genetic models which determined the extent to which lifetime depression symptoms, heart disease and hypertension shared genetic and/or environmental factors. RESULTS: Heart disease was significantly associated with 1 4 symptoms and 5 or more symptoms of depression (odds ratio [OR] = 2.62; 95% confidence interval [CI]: 1.54-4.46 and OR = 4.02; 95% CI: 2.16-7.46). Hypertension was significantly associated with 1 to 4 symptoms and 5 or more symptoms of depression (OR = 1.29; 95% CI: 1.11-1.50 and OR = 1.49; 95% CI: 1.21 1.83). The genetic correlations were significant between depression symptoms and hypertension (r =.19), and between depression symptoms and heart disease (r =.42). Of the total variance in depression, 8% was common to hypertension and heart disease, 7% of the variance in hypertension was common with depression symptoms and heart disease, and 64% of the variance in heart disease was common with depression symptoms and hypertension. CONCLUSIONS: Men who reported cardiovascular disease were significantly more likely to have depression symptoms. The lifetime co-occurrence of these phenotypes is partly explained by common genetic risk factors. PMID- 12883105 TI - Job strain and blood pressure in employed men and women: a pooled analysis of four northern italian population samples. AB - OBJECTIVE: The extent to which psychosocial stress concurs to raise blood pressure is still uncertain. Here the association between job strain and office blood pressure in a pooled analysis of four population samples from northern Italy is assessed. METHODS: Four surveys assessing prevalence of major coronary risk factors were performed in 1986, 1990, 1991, and 1993 in area "Brianza" (Milan), a World Health Organization-MONItoring cardiovascular disease (WHO MONICA) Project collaborating center. Ten year age- and gender-stratified independent samples were randomly recruited from the 25- to 64-year-old residents. The methods used to assess coronary risk factors strictly adhered to the MONICA manual, were kept constant, and underwent internal and external quality controls. Job strain was investigated through the administration to employed participants of a questionnaire derived from the Karasek model, assessing job demand/control latitude. Analysis was restricted to 25- to 54-year old participants, untreated for hypertension (1799 men and 1010 women). RESULTS: Among men, there was a 3 mm Hg increase of systolic blood pressure (p<.001) moving from low to high strain job categories. This difference was independent from age, education, body mass index, alcohol intake, smoking habits, leisure time physical activity, and survey. No relevant differences among job strain categories were found in women and for diastolic blood pressure in both gender groups. CONCLUSIONS: These results carried out on a large population-based sample confirm previous findings obtained adopting ambulatory blood pressure measurements in more restricted samples of population or patients. Further research is needed to clarify the relationship between perceived work stress and blood pressure in women. PMID- 12883106 TI - Alterations in brain and immune function produced by mindfulness meditation. AB - OBJECTIVE: The underlying changes in biological processes that are associated with reported changes in mental and physical health in response to meditation have not been systematically explored. We performed a randomized, controlled study on the effects on brain and immune function of a well-known and widely used 8-week clinical training program in mindfulness meditation applied in a work environment with healthy employees. METHODS: We measured brain electrical activity before and immediately after, and then 4 months after an 8-week training program in mindfulness meditation. Twenty-five subjects were tested in the meditation group. A wait-list control group (N = 16) was tested at the same points in time as the meditators. At the end of the 8-week period, subjects in both groups were vaccinated with influenza vaccine. RESULTS: We report for the first time significant increases in left-sided anterior activation, a pattern previously associated with positive affect, in the meditators compared with the nonmeditators. We also found significant increases in antibody titers to influenza vaccine among subjects in the meditation compared with those in the wait-list control group. Finally, the magnitude of increase in left-sided activation predicted the magnitude of antibody titer rise to the vaccine. CONCLUSIONS: These findings demonstrate that a short program in mindfulness meditation produces demonstrable effects on brain and immune function. These findings suggest that meditation may change brain and immune function in positive ways and underscore the need for additional research. PMID- 12883107 TI - Mindfulness-based stress reduction in relation to quality of life, mood, symptoms of stress, and immune parameters in breast and prostate cancer outpatients. AB - OBJECTIVES: This study investigated the relationships between a mindfulness-based stress reduction meditation program for early stage breast and prostate cancer patients and quality of life, mood states, stress symptoms, lymphocyte counts, and cytokine production. METHODS: Forty-nine patients with breast cancer and 10 with prostate cancer participated in an 8-week MBSR program that incorporated relaxation, meditation, gentle yoga, and daily home practice. Demographic and health behavior variables, quality of life (EORTC QLQ C-30), mood (POMS), stress (SOSI), and counts of NK, NKT, B, T total, T helper, and T cytotoxic cells, as well as NK and T cell production of TNF, IFN-gamma, IL-4, and IL-10 were assessed pre- and postintervention. RESULTS: Fifty-nine and 42 patients were assessed pre- and postintervention, respectively. Significant improvements were seen in overall quality of life, symptoms of stress, and sleep quality. Although there were no significant changes in the overall number of lymphocytes or cell subsets, T cell production of IL-4 increased and IFN-gamma decreased, whereas NK cell production of IL-10 decreased. These results are consistent with a shift in immune profile from one associated with depressive symptoms to a more normal profile. CONCLUSIONS: MBSR participation was associated with enhanced quality of life and decreased stress symptoms in breast and prostate cancer patients. This study is also the first to show changes in cancer-related cytokine production associated with program participation. PMID- 12883108 TI - Hostility, social support, and adrenergic receptor responsiveness among African American and white men and women. AB - OBJECTIVE: We examined the relationship between beta-adrenergic receptor responsiveness and hostility and social support in African American and white men and women. METHODS: The participants were 149 men and women, aged 25 to 45 years with SBP < 160 and DBP < 105. Hostility and social support were assessed with standardized self-report measures. An isoproterenol challenge was used to evaluate beta-adrenergic receptor responsiveness, and a phenylephrine challenge was used to evaluate alpha-adrenergic receptor responsiveness. RESULTS: Hostility and social support were unrelated to alpha-adrenergic receptor responsiveness. Hostility and satisfaction with perceived social support predicted beta adrenergic receptor responsiveness in multiple linear regression analyses controlling for race, gender, age, SBP, and resting heart rate. High hostility was associated with reduced cardiac beta-adrenergic receptor function among both white and African American men. Low levels of satisfaction with social support were associated with reduced cardiac beta-adrenergic receptor responsiveness among men and women. Hostility and satisfaction with social support shared some variance in models predicting beta-adrenergic receptor responsiveness. CONCLUSIONS: Reduced beta-adrenergic receptor responsiveness is associated with higher levels of hostility among men, and is associated with lower levels of satisfaction with social support among men and women. Impaired beta-adrenergic receptor function, which is a common characteristic of cardiovascular disease, may be a marker of increased cardiovascular disease risk among individuals high in hostility and low in social support. PMID- 12883109 TI - Expressive/suppressive anger-coping responses, gender, and types of mortality: a 17-year follow-up (Tecumseh, Michigan, 1971-1988). AB - OBJECTIVES: This study examined prospectively (1971-1988) the relationship between anger-coping responses, gender, and mortality (N = 91) in a representative sample of men (N = 324) and women (N = 372), aged 30 to 69, from the Tecumseh Community Health Study. METHODS: Anger-coping was measured by responses to hypothetical unfair anger-provoking situations. Cox proportional hazard regressions were used adjusted for seven health risk factors (age, smoking, relative weight, systolic blood pressure (SBP), bronchial problems, FEV1, and cardiovascular (CV) risk). RESULTS: Men's suppressed anger interacted significantly with SBP and also with bronchial problems to predict both all-cause and CV mortality. Women showed direct relationships between suppressed anger and early mortality (all-cause, CV, and cancer). Women also showed an interaction of spouse-suppressed anger and SBP for all-cause and CV mortality. Data suggest men who expressed their anger died earlier of cancer (N = 16) deaths. CONCLUSIONS: Suppressed anger at the time of an unjust attack may become chronic resentment (intermittent rage or hatred) about which little is known and requires research. The design for future research should experimentally measure both suppressed anger-coping responses (after an unfair attack) and morbidity (eg, blood pressure, bronchitis, immune disorder, etc.) to predict prospectively to earlier mortality. PMID- 12883110 TI - Acute psychological stress: effects on chemotaxis and cellular adhesion molecule expression. AB - OBJECTIVE: Activation of a psychological stress response increases autonomic activity and enhances immune function by inducing a significant increase in numbers of leukocytes at sites of inflammation. Chemotaxis and cellular adhesion are thought to mediate leukocyte trafficking. In this study, we examine the effects of an acute psychological stress on chemotactic responses of PBMCs and on CAM expression in relation to measures of sympathetic activation. METHODS: Subjects underwent either a public speaking task (N = 24) or a control condition (N = 13). Blood was drawn before the task, immediately after, and 20 minutes after, the task for changes in percentage of cells expressing cellular adhesion molecules, chemotaxis to chemokines, HR, blood pressure, and E and NE levels. RESULTS: In response to the laboratory stressor, increases of PBMC chemotaxis to FMLP and SDF-1 were found, which were coupled with increases in the percentages of lymphocytes expressing the integrin Mac-1. Autonomic activity, including blood pressure and circulating levels of catecholamines, increased after administration of the stressor, and correlated with increases of Mac-1. CONCLUSIONS: These data show that acute stress induces increase of chemotaxis and expression of CAM expression, which may contribute to increased migration and recruitment of immune cells to sites of infection and/or inflammation. PMID- 12883111 TI - Stress as a determinant of saliva-mediated adherence and coadherence of oral and nonoral microorganisms. AB - OBJECTIVE: The mucosal secretory proteins, such as the salivary proteins, play a key role in the acquisition and regulation of the mucosal microflora. Most notably, some microorganisms utilize the host's secretory proteins to adhere to the mucosa; a first step in colonization and infection. The secretory proteins also influence colonization by affecting the binding among microorganisms, a process denoted as coadherence. Previously we reported that acute stressors cause specific changes in saliva composition. The present study investigated to what extent these changes influence saliva-mediated microbial adherence and coadherence (ex vivo). METHODS: Thirty-two male undergraduates provided unstimulated saliva before and during a control condition and two stressors: A memory test and a surgery video presentation. We used saliva-coated microplates to test the adherence of bacteria for which the oral cavity is either a natural reservoir (eg, viridans streptococci) or a portal of entry (eg, Helicobacter pylori). We also tested the saliva-mediated co-adherence between Streptococcus gordonii and the yeast Candida albicans. Correlation analyses were performed to determine the relationships between changes in microbial adherence and the concentrations of potential salivary ligands, viz. cystatin S, the mucins MUC5B and MUC7, S-IgA, lactoferrin, alpha-amylase, and total salivary protein. RESULTS: During the memory test, saliva-mediated adhesion of Streptococcus sanguis, Streptococcus gordonii, and H. pylori increased, whereas the coadherence of C. albicans with S. gordonii decreased. During the surgical video presentation the saliva-mediated adherence of H. pylori, S. sanguis, and Streptococcus mitis increased. These changes were independent of salivary flow rate, but correlated with specific changes in salivary protein composition. CONCLUSION: The results show that even moderate stressors, by altering the activity of the mucosal secretory glands, may affect microbial colonization processes such as adherence and coadherence. This study hereby presents a mechanism by which stress may affect the mucosal microflora and susceptibility to infectious disease. PMID- 12883112 TI - Hemodynamics and arterial properties in response to mental stress in individuals with mild hypertension. AB - OBJECTIVE: The role of tonic sympathetic stimulation on the properties of large arteries is largely unknown. The purpose of this study was to determine the effect of mental stress on hemodynamics and arterial properties in mild hypertensives. METHOD: Twenty-three subjects with mild hypertension and 19 age matched normotensives were compared to examine changes in hemodynamics and central arterial wave reflection before, during, and after mental stress. RESULTS: The results demonstrate an acute effect of mental stress on blood pressure, heart rate, and arterial compliance. The static component (MBP) and the pulsatile (PP) component of arterial pressure increased significantly during mental stress and returned to baseline within a few minutes. Mild hypertensives did not have an increased response to mental stress. For both groups, an increase in HR and a consequent rise in CO were responsible for the increase in BP in response to mental stress. Compared with baseline, both groups demonstrated a decrease in arterial compliance during stress. Mental stress did not induce a significant change in total peripheral vascular resistance nor did it affect central arterial wave reflection in both groups. Individuals with mild hypertension demonstrated higher PP (p <.001), lower arterial compliance (p <.01), and higher AI (p <.05) than those with normal BP. CONCLUSIONS: Hemodynamic and arterial responses to mental stress in individuals with normal BP and mild hypertension were similar. Several parameters, however, were different in basal state. These differences (ie, higher PP, lower compliance, and higher AI in the mild hypertensive group) could be due to the chronic effect of sympathetic stimulation on central arteries. PMID- 12883113 TI - Positive affect predicts lower risk of AIDS mortality. AB - OBJECTIVE: The objective of this study was to test the association of positive affect as measured by the Center for Epidemiologic Studies Depression Scale (CES D) with risk of AIDS mortality, controlling for the other CES-D subscales and laboratory measures of disease progression. METHODS: Data come from the San Francisco Men's Health Study, a prospective study of a household probability sample of single men in San Francisco. The subjects were 407 men who were HIV+ at study baseline. RESULTS: In time-dependent Cox proportional hazards models, the positive affect subscale of the CES-D was significantly associated with lower risk of AIDS mortality (RR = 0.89, CI = 0.84-0.95). When risk estimates were adjusted for time-dependent covariates of CD4, serum beta2-microglobulin, P24 antigen, antiretroviral use, and the other subscales of the CES-D, positive affect remained significantly predictive of lower risk of AIDS mortality (RR = 0.90, CI = 0.85-0.97). When the CES-D subscale predictors were lagged by 12, 24, and 36 months in order to address the possibility that positive affect was simply a marker for better health, positive affect remained significantly predictive lagged by 12 months and marginally predictive lagged by 24 months. CONCLUSIONS: Positive affect seems to be the "active ingredient" in the association of scores on the CES-D depressive mood scale and mortality in this sample of HIV+ men. Future work should expand the traditional negative-affect-only focus to encompass the significant role that positive affect plays in living with HIV. PMID- 12883114 TI - Psychological distress is associated with decreased memory helper T-cell and B cell counts in pre-AIDS HIV seropositive men and women but only in those with low viral load. AB - OBJECTIVE: Although some studies have demonstrated the association of psychological distress and diminished immune system function in HIV spectrum disease, other studies have yielded apparently conflicting findings; the lack of consideration of the role of HIV viral burden may be central to this controversy. This study examined whether HIV viral burden moderated the relationship between psychological distress and enumerative and functional immune measures in pre-AIDS HIV spectrum disease. METHODS: This cross-sectional study used factor analysis to derive a composite measure of psychological distress incorporating measures of dysphoria, anxiety, and perceived stress. Multiple regression analyses used distress as the predictor, immune measures as the outcome variables, with viral load as the moderator variable, while controlling for age, medication use, and HIV symptomatology. Subjects were 148 pre-AIDS, HIV seropositive men and women (89 asymptomatic, 59 symptomatic), aged 18 to 45 years. The main outcome measures were enumerative and functional immune measures. RESULTS: A model of psychological distress was derived using each of the proposed measures. Findings indicated that high distress was associated with decreased numbers of helper T (memory) cells and B cells, but only at low levels of viral burden after controlling for age, medication use, and HIV-related symptoms. CONCLUSIONS: These findings highlight the importance of assessing the role of HIV viral burden when examining distress-immunity relationships in HIV-infected individuals. The lack of association in those persons with high viral load suggests that, even before AIDS onset, disease-related processes are disrupting CNS and immune system communication. PMID- 12883115 TI - Psychological distress and cancer survival: a follow-up 10 years after diagnosis. AB - OBJECTIVE: This study tested the predictive role of psychological distress in cancer survival, while attempting to overcome several important methodological and statistical limitations that have clouded the issue. METHODS: Measures collected on a range of emotional and cognitive factors in the early postdiagnostic period and at 4-month intervals up to 15 months after diagnosis were used to predict survival time up to 10 years among 205 cancer patients heterogeneous in disease site, status, and progression. RESULTS: With the use of both baseline and repeated measures, depressive symptomology was the most consistent psychological predictor of shortened survival time, after controlling for several known demographic and medical risk factors. CONCLUSIONS: Given the importance of depressive symptoms to cancer survival, discussion focuses on the possible mechanisms mediating this relationship, the importance of psychological screening of cancer patients, and need for further research. PMID- 12883116 TI - Psychosocial stress and cervical neoplasia risk. AB - OBJECTIVE: We assessed the association between psychosocial stress and preinvasive cervical neoplasia development controlling for HR-HPV infection. METHODS: This case-control study enrolled low-income women receiving family planning services at health department clinics. There were 59 cases with biopsy confirmed HSIL and 163 with low-grade SIL and 160 controls with normal cervical cytology. A modified SLE scale was used to measure stressful events and the perceived impact of the event in the prior 5 years. Unconditional logistic regression was used to assess SIL risk and stressful events scores and by subscales. RESULTS: After adjusting for age, HR-HPV infection, and lifetime number of sex partners, the SLE count score was associated with an increased risk of SIL among white women (aOR = 1.20; 95% CI = 1.04, 1.38) yet not among African American women (aOR = 1.02; 95% CI = 0.87, 1.19). The relationship stress subscale (divorce, infidelity, an increase in the number of arguments, and psychological and physical partner violence) was the only one of four subscales (loss, violence, and financial stress) associated with SIL, again, only among white women (aOR = 1.54; 95% CI = 1.21, 1.96). CONCLUSIONS: These data suggest that psychosocial stress may play a role in SIL development. Future studies are needed to confirm these findings, to explore racial difference in reporting stress, and to explore the mechanism through which psychosocial stress may affect cervical neoplasia risk. PMID- 12883117 TI - Emotional style and susceptibility to the common cold. AB - OBJECTIVE: It has been hypothesized that people who typically report experiencing negative emotions are at greater risk for disease and those who typically report positive emotions are at less risk. We tested these hypotheses for host resistance to the common cold. METHODS: Three hundred thirty-four healthy volunteers aged 18 to 54 years were assessed for their tendency to experience positive emotions such as happy, pleased, and relaxed; and for negative emotions such as anxious, hostile, and depressed. Subsequently, they were given nasal drops containing one of two rhinoviruses and monitored in quarantine for the development of a common cold (illness in the presence of verified infection). RESULTS: For both viruses, increased positive emotional style (PES) was associated (in a dose-response manner) with lower risk of developing a cold. This relationship was maintained after controlling for prechallenge virus-specific antibody, virus-type, age, sex, education, race, body mass, and season (adjusted relative risk comparing lowest-to-highest tertile = 2.9). Negative emotional style (NES) was not associated with colds and the association of positive style and colds was independent of negative style. Although PES was associated with lower levels of endocrine hormones and better health practices, these differences could not account for different risks for illness. In separate analyses, NES was associated with reporting more unfounded (independent of objective markers of disease) symptoms, and PES with reporting fewer. CONCLUSIONS: The tendency to experience positive emotions was associated with greater resistance to objectively verifiable colds. PES was also associated with reporting fewer unfounded symptoms and NES with reporting more. PMID- 12883118 TI - Hopelessness is associated with decreased heart rate variability during championship chess games. AB - OBJECTIVE: Clinical observations suggest that negative affects such as helplessness/hopelessness (HE/HO) may induce autonomic duration; affects were assessed for every move after reconstruction of the games. In all games compiled, 18 situation of intense confidence/optimism and 20 of intense helplessness/hopelessness were observed. RESULTS: Intense affects of HE/HO were associated with decreasing HF-HRV (Fisher exact test, p =.003), increasing "nervousness" (p =.0005), decreasing "optimism" (p =.0005), and decreasing "calmness" (p =.0005). CONCLUSIONS: Investigation of championship chess game players with an ELO strength > or = 2300 in a natural field setting revealed increasing HE/HO being associated with reduced HF-HRV suggestive of vagal withdrawal. Thus, our data may help link negative mood states, autonomic nervous system disturbances, and cardiac events. PMID- 12883119 TI - Apolipoprotein E genotype and temperament: a longitudinal study from infancy to the late teens. AB - OBJECTIVE: To replicate an earlier Finnish study by Keltikangas-Jarvinen et al. (5) reporting that the APOE genotype is associated with temperamental traits involving increased activity. METHODS: DNA was collected from 683 Australian children who had participated in a longitudinal study of childhood temperament from 4 to 8 months up to 17 to 18 years. Associations were examined between APOE genotype and a range of measures of activity and hyperactivity. RESULTS: No associations were found. CONCLUSIONS: The earlier Finnish finding could not be replicated despite adequate statistical power. PMID- 12883120 TI - Tryptophan as a link between psychopathology and somatic states. AB - OBJECTIVE: Several somatic illnesses are associated with psychiatric comorbidity. Evidence is provided that availability of the essential amino acid tryptophan, which is the precursor of serotonin, may cause this phenomenon. METHODS: We performed a database search to find relevant articles published between 1966 and 2002. For our search strategy, we combined several diseases from the categories hormonal, gastrointestinal, and inflammatory with the search terms "tryptophan" and "serotonin." RESULTS: The catabolism of tryptophan is stimulated under the influence of stress, hormones and inflammation by the induction of the enzymes tryptophan pyrrolase (in the liver) and IDO (ubiquitous). Because of the reduction in blood levels of tryptophan under these circumstances the formation of cerebral serotonin is decreased. CONCLUSIONS: It is argued that the coupling of peripheral tryptophan levels and cerebral serotonin levels has physiological significance. The clinical implications and therapeutic consequences of changes in tryptophan and consequently serotonin metabolism are discussed. PMID- 12883121 TI - Reduced glucocorticoid sensitivity of monocyte interleukin-6 production in male industrial employees who are vitally exhausted. AB - OBJECTIVE: Proinflammatory changes are thought to link vital exhaustion with adverse cardiovascular outcomes. Monocytes play a central role in the pathogenesis of atherosclerotic lesions and are a major source of circulating cytokines. We hypothesized that vital exhaustion may alter the regulation of monocyte activity, as measured by lipopolysaccharide (LPS)-stimulated and glucocorticoid inhibited release of the proinflammatory cytokine interleukin-6 (IL-6). METHODS: In 166 middle-aged apparently healthy men, vital exhaustion was measured by the Shortened Maastricht Exhaustion Questionnaire. Subjects in the highest quartile (highly exhausted, N= 38) were compared with those in the second and third quartiles (moderately exhausted N= 89) vs. those in the lowest quartile (nonexhausted, N= 39) in terms of plasma C-reactive protein (CRP) and tumor necrosis factor-alpha (TNF-alpha) levels, and as to IL-6 release after LPS stimulation in vitro. Inhibition of IL-6 release was determined by coincubation with increasing concentrations of dexamethasone. Monocyte glucocorticoid sensitivity was defined as the dexamethasone concentration inhibiting IL-6 release by 50%. RESULTS: Highly exhausted individuals had higher CRP levels than nonexhausted subjects (p=.008). LPS-stimulated IL-6 release was not significantly different between groups. However, in highly exhausted participants, dexamethasone was less able to inhibit IL-6 release (p=.010), and the glucocorticoid sensitivity was lower (p=.003) than in nonexhausted subjects. CONCLUSIONS: In highly exhausted individuals, glucocorticoids exert less suppressive action on monocyte IL-6 release than in nonexhausted subjects. This finding points to altered regulation of monocyte cytokine production as one possible pathway linking exhaustion with atherosclerosis. PMID- 12883122 TI - Salivary cortisol response during exposure treatment in driving phobics. AB - OBJECTIVE: Extensive research on the hypothalamic-pituitary-adrenal (HPA) axis response to stress has not clarified whether that axis is activated by phobic anxiety. We addressed this issue by measuring cortisol in situational phobics during exposure treatment. METHODS: Salivary cortisol was measured in 11 driving phobics before and during three exposure sessions involving driving on crowded limited-access highways and compared with levels measured in 13 healthy controls before and during two sessions of driving on the same highways. For each subject, data collected in the same time period on a comparison nondriving day served as an individual baseline from which cortisol response scores were calculated. RESULTS: Cortisol levels of driving phobics and controls did not differ on the comparison day. Phobics also had normal cortisol response scores on awakening on the mornings of the exposures but these were already increased 1 hour before coming to the treatment sessions. Phobics had significantly greater cortisol response scores during driving exposure and during quiet sitting periods before and afterward. These greater responses generally paralleled increases in self reported anxiety. At the first exposure session, effect sizes for differences in cortisol response scores between the two groups were large. Initial exposure to driving in the first session evoked the largest responses. CONCLUSION: The data demonstrate that the HPA axis can be strongly activated by exposure to, and anticipation of, a phobic situation. PMID- 12883123 TI - Social networks and marital status predict mortality in older women: prospective evidence from the Study of Osteoporotic Fractures (SOF). AB - OBJECTIVE: To assess the relationship between social network size and prospective mortality risk among a large sample of older, Caucasian women. METHOD: The study included 7524 Caucasian community-dwelling women, age 65 or older (mean age = 74.1), who participated from four U.S. communities. Study participants completed a protocol that included anthropomorphic and health assessments at baseline and the Lubben Social Network Scale at year 2. We followed participants for an average of 6 years after they had completed the year-2 assessment. We used hospital records and a copy of the participant's official death certificate to document mortality and cause of death in accordance to ICD-9 revision codes. RESULTS: A total of 1451 deaths (19.3% of sample) were observed over follow-up, 215 (3.4%) due to cardiovascular causes. Higher social network scores were a robust predictor of lower multivariate-adjusted mortality (RR = 0.92, 95% CI = 0.86-0.98), controlling for age, comorbid disease, body mass, smoking, depression, and education. However, social network benefits were attenuated after controlling for marital status. Married participants showed lower total (RR = 0.83, 95% CI = 0.74-0.94) and CVD (RR = 0.59, 95% CI = 0.43-0.81) covariate adjusted death rates compared with unmarried participants. CONCLUSIONS: Social network scores and marriage were each associated with reduced prospective mortality risk among older women. The relationships shown here suggest that much of the protection afforded by larger social networks in older women results from marriage rather than other forms of social relationships. Mechanisms at the physiological or behavioral level explaining social relationship benefits remain important areas for future research. PMID- 12883124 TI - Childhood family dysfunction and associated abuse in patients with nonepileptic seizures: towards a causal model. AB - OBJECTIVE: A history of childhood sexual abuse is thought to characterize patients with nonepileptic seizures (NES). We tested the hypotheses: 1) that history of sexual abuse is more prevalent in patients with NES than in controls with epilepsy; 2) that such abuse is associated with NES, not directly but because it is a marker of family dysfunction; and 3) that family dysfunction and abuse are, in turn, linked to NES because they increase a general tendency to somatize. METHODS: We compared 81 patients with NES with 81 case-matched epilepsy patients, using questionnaires to elicit recollections of sexual, physical, and psychological abuse and family atmosphere and to quantify current somatization. RESULTS: Although each form of abuse was more prevalent in NES patients, only child psychological abuse uniquely distinguished NES from epilepsy. However, its association with NES was explained by family dysfunction. A general tendency to somatize explained part of the relationship of abuse to NES. CONCLUSIONS: Abuse therefore seems to be a marker for aspects of family dysfunction that are associated with--and may therefore cause--somatization and, specifically, NES. PMID- 12883125 TI - The spectrum of eating disorders in young women: a prevalence study in a general population sample. AB - OBJECTIVE: The present study aims to evaluate the prevalence and characteristics of the whole spectrum of eating disorders (ED) in a representative sample of young women. METHOD: All female subjects aged 18 to 25 who resided in two areas (urban and suburban) of a large city were involved in the study. All women (N = 934) underwent a clinical interview which included the structured clinical interview for DSM-IV. RESULTS: Lifetime anorexia nervosa (AN) and bulimia nervosa (BN) were diagnosed respectively in 2.0% and 4.6% of the subjects. The prevalence of lifetime atypical ED was 4.7% and that of binge eating disorder (BED) was 0.6%. The degree of urbanization has a significant impact on the prevalence of AN, BN, and BED. Social class, professional status, and education were not associated with an increased risk of reporting an ED, whereas the number of hypocaloric diets, having been a victim of childhood abuse, and, in BN, ever being overweight are significantly associated with ED. CONCLUSIONS: Our findings have confirmed the importance of community studies to improve our knowledge about factors that have some influence on pathogenesis, treatment referral, and outcome. PMID- 12883126 TI - Cognitive changes predict continued recovery of erectile functioning versus relapse after discontinuation of sildenafil treatment for male erectile dysfunction. AB - OBJECTIVE: To examine whether erectile functioning after termination of sildenafil intake in men with psychogenic ED (erectile dysfunction) can be predicted with psychological measures. METHOD: The subjects in a nonrandomized controlled trial were 65 heterosexual men with acquired psychogenic ED, aged 54.2 +/- 11 years. Sildenafil medication was taken as required before sexual activity, up to two times per week. Response to a global end point question ("Did the treatment you took during the study improve your physical response during sexual activity in the last 4 weeks?") was recorded after 6 weeks of sildenafil use and, subsequently, 6 weeks without medication. Other measures of sexual functioning and cognitive predictor measures were also administered. RESULTS: Of the 65 participants who commenced sildenafil treatment, 37% withdrew from the study before follow-up assessment. At posttreatment, 89% of participants reported that treatment had improved or cured their erectile functioning. At follow-up, 66% of participants had maintained posttreatment gains. Response at follow-up could be predicted (p <.001) with 96% sensitivity and 50% specificity by entering changes in sexual self-confidence and the participant's rating of his partner's wish to continue treatment in a logistic regression model. Higher odds for recovery of erectile functioning were found in participants reporting increased sexual self confidence and the estimation that their partner wanted them to continue sildenafil use. High pretreatment sexual desire was found to further increase the odds for positive responding at follow-up. CONCLUSIONS: The present results indicate that continued improvement of erectile functioning is possible after discontinuation of sildenafil use in men with psychogenic ED. Maintenance of gains can be predicted from cognitive changes before medication is withdrawn. PMID- 12883127 TI - Bupropion in psoriasis and atopic dermatitis: decreased tumor necrosis factor alpha? PMID- 12883128 TI - Managed care or ethical care: what's in a name? PMID- 12883130 TI - Underuse of evidence-based pharmacotherapies for affective disorders. PMID- 12883131 TI - Developing ethical guidelines for pharmaceutical company support in an academic mental health center. PMID- 12883132 TI - Antipsychotic polypharmacy in the ambulatory care setting, 1993-2000. PMID- 12883133 TI - The best of practices, the worst of practices: The making of science-based primary prevention programs. PMID- 12883134 TI - The costs of treating persons with depression and alcoholism compared with depression alone. PMID- 12883135 TI - Emergency psychiatry and its vicissitudes. PMID- 12883136 TI - Client versus client. PMID- 12883137 TI - Perceptions of discrimination among persons with serious mental illness. AB - OBJECTIVES: The authors sought to gain further perspective on discrimination experienced by persons with mental illness by comparing self-reports of discrimination due to mental illness to self-reports of discrimination due to other group characteristics, such as race, gender, and sexual orientation. METHODS: A total of 1,824 persons with serious mental illness who participated in a baseline interview for a multistate study on consumer-operated services completed a two-part discrimination questionnaire. The first part of the questionnaire assessed participants' perceptions about discrimination due to mental illness as well as more than half a dozen other group characteristics. The second part of the questionnaire asked participants who reported some experience with discrimination to identify areas in which this discrimination occurred, such as employment, education, and housing. RESULTS: More than half of the study participants (949 participants, or 53 percent) reported some experience with discrimination. The most frequent sources of this discrimination were mental disability, race, sexual orientation, and physical disability. Areas in which discrimination frequently occurred included employment, housing, and interactions with law enforcement. Areas in which discrimination was experienced did not significantly differ among groups of study participants characterized by mental disability, race, gender, sexual orientation, or physical disability. CONCLUSIONS: Discrimination based on group characteristics other than mental illness does not diminish the impact of stigma associated with mental illness. Antistigma programs need to target not only discrimination related to mental illness but also that associated with other group characteristics, such as race, gender, sexual orientation, and physical disability. PMID- 12883138 TI - A review of pharmacologic treatments for obsessive-compulsive disorder. AB - OBJECTIVE: Obsessive-compulsive disorder is a chronic and often disabling disorder that affects 2 to 3 percent of the U.S. population. Optimal treatment involves a combination of pharmacologic and cognitive-behavioral therapies. Advances in psychopharmacology have led to safe and effective treatments for obsessive-compulsive disorder that provide clinically significant improvement in symptoms. In this article the authors review studies of pharmacologic treatments. METHODS: A MEDLINE search was conducted to identify relevant articles from 1991 to 2002. Double-blind, placebo-controlled studies as well as open-label studies and case reports were included. RESULTS AND DISCUSSION: The serotonin reuptake inhibitors (SRIs), including clomipramine, fluvoxamine, fluoxetine, sertraline, and paroxetine, have been approved by the U.S. Food and Drug Administration for the treatment of adults with obsessive-compulsive disorder; three of these (clomipramine, fluvoxamine, and sertraline) have been approved for treatment of children and adolescents. Clomipramine and the selective serotonin reuptake inhibitors (SSRIs) are first-line agents. However, 40 to 60 percent of patients with obsessive-compulsive disorder do not respond to adequate treatment trials with SRIs, and agents that alter serotonin receptors and other neurotransmitter systems, such as dopamine, norepinephrine, and second-messenger systems, may play a role in treatment. Treatment options for patients who do not respond to SRIs include switching, augmentation, or novel-agent strategies. Up to two-thirds of patients with obsessive-compulsive disorder have comorbid psychiatric disorders, which may present a challenge in pharmacologic treatment. Major depressive disorder is the most common comorbid condition. Nonpharmacologic invasive techniques may play a role in refractory cases of obsessive-compulsive disorder, but further research is warranted. PMID- 12883139 TI - An analysis of successful efforts to reduce the use of seclusion and restraint at a public psychiatric hospital. AB - OBJECTIVE: The author reviews and evaluates a variety of interventions that were considered to have contributed to the successful reduction of reliance on the use of seclusion and restraint in a public psychiatric hospital for adult patients with severe and persistent psychiatric impairments. METHODS: A multiple regression analysis was applied to monthly seclusion and restraint data covering the period from 1997 to 2002. The independent variables were each of the component efforts to reduce reliance on seclusion and restraint at the hospital: changes in the criteria for administrative review of incidents of seclusion and restraint, changes in the composition of the case review committee, development of a behavioral consultation team, enhancement of standards for behavioral assessments and plans, and improvements in the staff-patient ratio. RESULTS: The use of seclusion and restraint was 75 percent lower during the final year of the five-year study period than it was during the first year. The only variable that was significantly associated with reduction in the use of seclusion and restraint was changes in the process for identifying critical cases and initiating a clinical and administrative case review. CONCLUSION: The results of this analysis underscore the importance of clinical and administrative priorities in efforts to reduce the use of seclusion and restraint in public psychiatric hospitals. PMID- 12883140 TI - Preferences for schizophrenia treatment outcomes among public policy makers, consumers, families, and providers. AB - OBJECTIVE: This study measured state public policy makers' ratings of the importance of several key schizophrenia treatment outcomes and compared them with the ratings of primary stakeholders in schizophrenia treatment. METHODS: Three groups of policy makers (40 administrative decision makers, 40 state legislators, and 20 legislative aides) and three groups of core stakeholders (20 persons with schizophrenia, 13 of their family members, and 20 of their mental health care providers) were recruited in Florida. Participants rated 12 descriptions of schizophrenia-related health states that reflected better and worse outcomes in six domains: psychotic symptoms, deficit symptoms, medication side effects, productive activity, daily activity, and social activity. RESULTS: All participants valued functional outcomes, such as improved productive and social activity, more than they valued improvements in symptoms. Public policy makers and primary stakeholders differed in the value they placed on two of the six outcomes. Compared with primary stakeholders, policy makers valued improvements in social functioning significantly more and improvements in medication side effects significantly less. CONCLUSIONS: Policy makers and primary stakeholders place similar value on some of the major goals of schizophrenia treatment, with both groups valuing functional outcomes most highly. However, the difference between groups in the importance placed on medication side effects may lead to conflicts in the allocation of resources to the provision of newer and more expensive medications, which are associated with fewer side effects. This initial examination of policy makers' views provides a starting point for developing consensus about schizophrenia treatment policies. PMID- 12883141 TI - Cognitive and symptom predictors of work outcomes for clients with schizophrenia in supported employment. AB - OBJECTIVES: This study examined the relationships of measures of cognitive functioning and psychiatric symptoms with work outcomes and use of vocational services for clients with schizophrenia in a supported employment program. METHODS: Thirty clients who were newly enrolled in a supported employment program were evaluated with cognitive and symptom measures at program entry and two years later. The clients' amounts of competitive work, wages earned, on-job support, and contact with employment specialists during the two-year follow-up period were documented. RESULTS: Predictors of clients' work outcomes included previous work history, amount of government entitlement income received, severity of negative symptoms, involvement in sheltered work activity at baseline, and level of cognitive functioning, including scores on measures of executive functioning and verbal learning and memory. The amounts of on-job support and contact with employment specialists were predicted by the cognitive domains of executive functioning, verbal learning, attention, and psychomotor speed as well as by the severity of psychotic symptoms. CONCLUSIONS: Clients with schizophrenia who have higher levels of cognitive impairment may require greater amounts of vocational support than those with lower levels of impairment. A variety of rehabilitation strategies may be required to improve vocational outcomes and reduce the amount of supported employment services needed by clients with schizophrenia. PMID- 12883142 TI - Characteristics of third-party money management for persons with psychiatric disabilities. AB - OBJECTIVE: The study examined different types of third-party money management arrangements for persons with psychiatric disabilities and consumers' perceptions of their finances in the context of these arrangements. METHODS: Clinical and demographic data were collected through structured interviews and record reviews for 240 persons with a diagnosis of a psychotic or major affective disorder who had been involuntarily hospitalized and were awaiting discharge on outpatient commitment in North Carolina. All consumers were receiving Supplemental Security Income or Social Security Disability Insurance. RESULTS: Third-party money management arrangements were reported by 102 (41 percent) of the study participants. A majority (77 percent) of these consumers had their finances managed by a family member. Consumers with third-party money managers were more likely to have a median annual income below 5,000 US dollars, to have a diagnosis of a primary psychotic disorder, and to have substance use problems. Most participants with third-party money managers reported that they received sufficient money to cover basic expenses, although about half also perceived having insufficient money to participate in enjoyable activities. CONCLUSIONS: Given that treatment for severe mental illness emphasizes social skills training and development of social support networks, financial limitations could undermine therapeutic efforts. It is important that clinicians consider the role of financial concerns when assessing consumers. Additional research should be conducted to better understand the role of financial variables in providing effective mental health services. PMID- 12883143 TI - Gambling participation and problems among South East Asian refugees to the United States. AB - OBJECTIVE: Gambling is common among South East Asian refugees, but no known studies have evaluated the prevalence of pathological gambling in these populations. The purpose of this study was to assess rates of gambling participation and gambling problems among South East Asian refugees. METHODS: Ninety-six immigrants to the United States from Laos, Cambodia, and Vietnam who attended community service organizations for these ethnic groups in Connecticut were asked to complete the South Oaks Gambling Screen (SOGS), which had been translated into their native languages. Demographic information as well as data on recent gambling activities were also obtained. RESULTS: The SOGS retained high internal consistency in the sample, with a Cronbach's alpha of .90. The lifetime prevalence of pathological gambling was 59 percent. Rates of gambling problems did not differ across the three ethnic groups. However, being male, divorced or separated, and younger were significant predictors of pathological gambling. More than half of all the respondents had gambled within two weeks of the interview, and 42 percent had wagered more than 500 US dollars in the previous two months. CONCLUSIONS: These data call for more research into the social, environmental, and cultural context of gambling among South East Asian refugees. Ethnically sensitive prevention and intervention strategies are needed to address the extraordinarily high rates of gambling problems in this population. PMID- 12883144 TI - Factors associated with high use of public mental health services by persons with borderline personality disorder. AB - OBJECTIVE: The research presented here was a pilot study to identify clinical factors associated with high use (as opposed to lower use) of inpatient psychiatric services by persons with borderline personality disorder. METHODS: The initial sample was a random sample of English- and Spanish-speaking persons aged 18 to 60 years who had received at least one outpatient mental health service in the previous 90-day period and were enrolled in one of the participating mental health centers in King County, Washington. A random sample of persons who met selection criteria was randomly drawn; persons with high levels of use were oversampled to ensure adequate representation. Twenty-nine participants met full criteria for borderline personality disorder on the Personality Disorders Examination structured interview and completed all measures. Fifteen (52 percent) of these had a high level of use of inpatient services, and 14 did not. RESULTS: High use of inpatient psychiatric services was predicted by a history of parasuicide in the previous two years but not by the number or severity of parasuicides; by the presence and number of anxiety disorders but not by depression or psychotic or substance use disorders; and by poorer cognitive functioning. Life stressors, global functioning, and health service variables did not differentiate patients with high levels of service use from other patients with borderline personality disorder. CONCLUSIONS: Further research should explore these predictors of service use to determine whether they are replicated in larger samples, and treatments that target these variables should be evaluated. PMID- 12883145 TI - A program for managing weight gain associated with atypical antipsychotics. AB - This study assessed the efficacy of a weight control program for patients taking atypical antipsychotics. Thirty-one patients with schizophrenia or schizoaffective disorder participated in a 12-week weight control program that incorporated nutrition, exercise, and behavioral interventions. Changes in patients' weight and in body mass index (BMI) were recorded and compared with those of 15 patients in a control group. The intervention group had a mean weight loss of 2.7 kg (six pounds) and a mean reduction of.98 BMI points, compared with a mean weight gain of 2.9 kg (6.4 pounds) and a mean gain of 1.2 BMI points in the control group. These data suggest that the intervention was effective in this group of patients. Professionals treating persons who are taking atypical antipsychotics should encourage them to engage in weight control activities. PMID- 12883146 TI - Barriers to primary medical care among patients at a community mental health center. AB - People with chronic mental disorders may be at risk of reduced access to medical treatment and poor quality of medical care. The authors examined receipt of and barriers to medical treatment among patients at a community mental health center. Fifty-nine patients completed a validated instrument for measuring access to medical care and the quality of primary care. Their responses indicated problems with access to care, and their scores were significantly lower than those of the general population on all four domains used to assess the quality of primary medical care. The findings suggest the need for community mental health centers to better assess and address barriers to primary medical care. PMID- 12883147 TI - Use of self-help services and consumer satisfaction with professional mental health services. AB - This study tested the hypothesis that users of mental health self-help services would be more satisfied with professional mental health services than clients who did not use self-help services. A survey was administered to 311 clients of professional mental health services, 151 (49 percent) of whom were users of self help services. A multiple regression model showed that the use of self-help services was associated with greater satisfaction with professional mental health services. This finding provides support for the idea that the use of self-help services encourages appropriate use of professional services. The study provided evidence that self-help and traditional mental health services can function complementarily rather than in competition with one another. PMID- 12883148 TI - Medication errors in psychiatry. PMID- 12883149 TI - Medication errors in psychiatry. PMID- 12883150 TI - Psychiatry in La-La Land. PMID- 12883152 TI - Use of oxcarbazepine for treatment-resistant aggression. PMID- 12883154 TI - Reliability of information provided by patients of a virtual psychiatric clinic. PMID- 12883155 TI - Initial care seeking for panic attacks. PMID- 12883156 TI - The wound infection continuum and its application to clinical practice. AB - Four basic conditions exist in open wounds resulting from the level of bioburden present (bacterial contamination--normal but short-lived state, colonization- normal state, critical colonization--abnormal state, and infection--abnormal state). The two abnormal states have the potential to disrupt the orderly healing sequence, which results in the development of a chronic wound. In addition, the impact of the wound s anatomical position, duration, shape, or presentation; the patient s level of health and control of underlying pathologies; the presence of infection-potentiating factors such as foreign bodies, hematoma, and necrotic tissue; the sources and frequency of exogenous contamination; the considered virulence of the individual micro-flora species; and the potential synergism between different species all require consideration. This article discusses the states of bioburden and other aspects of microbiology relating to wound infection and their application to clinical practice. PMID- 12883157 TI - Bacterial toxins and wound healing. AB - Infection--a factor commonly related to delayed closure for many chronic and acute wounds--is associated with relatively high levels of bacteria in viable tissues. But bacteria also can delay wound healing at lower levels before tissue invasion via toxin secretion either directly from viable cells (exotoxins) or as a result of cell lysis (endotoxins). These toxins tend to cause local necrosis and disrupt the delicate balance of critical mediators such as cytokines and proteases necessary for healing progression. Therefore, toxin control or absorption is a potentially valuable adjunct to any infection control modality. A long-standing method of controlling toxins of many types is the use of high surface area adsorbents, such as activated charcoal. Recent data suggest that activated charcoal may offer specific advantages in topical wound management through its effects on bacterial toxins. PMID- 12883158 TI - Pressure ulcers and infection--understanding clinical features. PMID- 12883159 TI - Bacterial swabs and the chronic wound: when, how, and what do they mean? PMID- 12883160 TI - Topical antimicrobials. PMID- 12883161 TI - A scientific perspective on the use of topical silver preparations. PMID- 12883162 TI - Systemic antimicrobial therapies for pressure ulcers. PMID- 12883163 TI - Wound bed preparation: future approaches. PMID- 12883168 TI - A novel magnetic resonance-positive emission image registration based on morphology. AB - Structures were obtained from images with morphological characteristics to register with voxel-based method. We applied simple morphological operations to obtain human brain cortex and chose normalized mutual information as similarity measure for the geometric alignment of PET and MR images. Evaluation used nine patients, and the results showed that sub-voxel accuracy was achieved and the registration process was significantly more rapid. Thus this new automated multi modality registration method is more robust and has high accuracy. PMID- 12883169 TI - Self-setting kinetics of new type calcium phosphate bioactive bone cement: a thermokinetics study. AB - Thermokinetics method was used to study the self-setting kinetics of a new kind of calcium phosphate cement (CPC) in the present study. A calcium-deficient hydroxyapatite CPC was developed by using alpha-TCP and other calcium phosphate bioceramics. The mixing liquids used were deionized water and 0.25 M NaH2PO4/Na2HPO4, respectively. The calorimetric curves, heat evolution curves and total heat evolution in the setting and hardening process of CPC were determined. It has been found that mixing liquids, reaction temperature had influences on the calorimetric curves and heat evolution, and mixing liquids exhibited the greatest influence on the kinetics of CPC during the self-setting and hardening process. Based on the calorimetric curves obtained, the kinetic model equation was simulated, and the reaction control step was determined. PMID- 12883170 TI - Tribological behavior of artificial hip joint under the effects of magnetic field in dry and lubricated sliding. AB - In recent years, there is an increasing utilization and demand to use magnetic fields in bioengineering applications due to its beneficial effects. Although in the last decade more attention has been given by tribologists to the electromagnetic processes taking place between sliding surfaces, which influence the tribological behaviors, but no attention has been concern with the sliding surfaces of the artificial implant joints. Therefore, the present work aims to elucidate the tribological behavior of an artificial joint implant under the effect of magnetic fields. Experimental investigation was carried out on a specially designed and constructed hip simulator on which the variations in the coefficients of friction and wear rates of the sliding surfaces were evaluated under the influence of a medium strength magnetic field suitable to apply in the human body. A realistic Ti-alloy implanted stem was used with an inserted head made from surgical grade stainless steel. This head was allowed to rub against UHMWPE sockets. The utilized type of prosthesis was "The JRI Modular Muller Standard-Total Hip Design". The performed experimental tests were conducted under both dry and lubricated sliding conditions using physiological saline solution. The designed simulator allows the coefficients of friction and the wear rates to be evaluated under realistic physiological loading and motion cycles encountered during normal walking of the human body. Comparative results are presented between the artificial joint performance in the presence and absence of the applied magnetic field. The experimental results have indicated that the presence of a medium strength magnetic field of 270 Gauss strength between rubbing surfaces resulted in high beneficial reductions in friction and wear rate of UHMWPE sliding on stainless steel either under dry or saline lubricating conditions. Therefore recommendation was forward to subject artificial implants made of stainless steel/UHMWPE combination of material to such medium strength magnetic field in animal clinical trials aiming to prolong the implant life. Scanning investigation of rubbing surfaces has revealed that the transfer of polymer to the counterface plays a dominant role in dictating the frictional and wear behaviors under dry sliding condition. Smooth molecular profile of the polymer-transferred leads to progressive reductions in friction and wear while the lumpy polymer transfer, formed at the beginning of sliding, increases both friction coefficient and wear. Two action mechanisms dominate the sliding process; adhesive and abrasive mechanisms. The presence of saline lubricant retards the formation of the beneficial polymer transfer thus leading to faster abrasion of the polymeric counterface which explains the relatively rapid and progressive increases in friction and wear. PMID- 12883171 TI - Creating bioabsorbable Schwann cell coated conduits through tissue engineering. AB - Enormous effort has been devoted to the generation of a synthetic guidance conduit for nerve repair instead of utilizing autograft. Several studies show neural guidance conduit is more effective when coated with Schwann cells. In this study, we synthesized bioabsorbable conduit consist of L-lactide and epsilon caprolactone which was useful clinically and examined adhesion of Schwann cells to bioabsorbable conduits. In vivo studies were done in which these polymer conduits coated with Schwann cells were implanted across a 12 mm gap in the rat sciatic nerve. Silicone conduits were implanted across the same gap as control. At 12 weeks, axonal regeneration was observed in the midconduit region of these polymer conduits and was not in control. This study assesses the feasibility of a tissue engineering approach to constructing bioabsorbable conduits coated with Schwann cells. PMID- 12883172 TI - The effect on wear of lift-off in total knee arthroplasty. AB - Femoral lift-off has been reported in total knee arthroplasty using dynamic fluoroscopic analysis. However, the effect of femoral lift-off on ultra-high molecular-weight polyethylene (UHMWPE) is not clear. The objective of this study was to evaluate the effect of lift-off movement on the wear of UHMWPE using a pin on-disc testing machine. Using oxidised UHMWPE, delamination was observed in 70% of the discs with lift-off movement, as against none in without lift-off movement (p=0.0603). In non-oxidised UHMWPE the depth of the wear track was increased in the impact area after lift-off, but there was no delamination and no increase in wear as measured by weight loss. UHMWPE can sustain the impact of after lift-off with neither delamination nor wear by other mechanisms. In contrast, degraded (i.e., oxidised) UHMWPE undergoes delamination after lift-off. Since it is probable that lift-off cannot be prevented, oxidised UHMWPE should never be used in TKA. PMID- 12883173 TI - Non-linear material models for tracheal smooth muscle tissue. AB - The aim of this study is to investigate the hyperelastic material models to describe the non-linear stress-strain behavior of tracheal smooth muscle tissue. Specifically, the goal is to validate the material model with experimental data using different finite element models and discuss the trends in stress-strain behavior of smooth muscle tissue. Both 2D and 3D finite element analyses were carried out to estimate the stress-strain behavior of the smooth muscle tissue. The results obtained indicate that the developed Ogden material model is valid and useful in explaining the stress strain behavior of tracheal smooth muscle tissue under different conditions. Finite element simulation results of the stress-strain behavior in the transverse direction are presented. PMID- 12883174 TI - Evaluation of apatite ceramics containing alpha-tricalcium phosphate by immersion in simulated body fluid. AB - The purpose of this study was to estimate the availability of alpha-tricalcium phosphate (alpha-TCP) on/in hydroxyapatite (HAP) ceramics for bioactivity as bone substitute materials by immersion in a simulated body fluid (SBF; Hanks' solution) containing ion concentrations similar to those in human blood plasma. Two alpha-TCP-surface-modified HAP and alpha-TCP-HAP composite materials were prepared by orthophosphoric acid treatment of sintered HAP and controlling the crystal phases of calcium phosphate cement, respectively. After immersion in SBF, the sintered HAP modified on the surface in an approximately 0.2 microm alpha-TCP layer was more effective for the precipitation of carbonated apatites than an approximately 2 microm alpha-TCP layer and HAP-only layer. In the calcium phosphate cements consisting of HAP and alpha-TCP phases, after immersion for 1 week, the specimens precipitated large amounts of apatites having alpha-TCP contents of approximately 25% and 50% in the cement. The results of immersion tests imply the possibility that the alpha-TCP on/in HAP ceramics may be a bioactive agent for bone-substituting HAP materials. PMID- 12883175 TI - Cement bond strengths of titanium plates. AB - The success of the oral rehabilitation of implant patients depends not only on the osseointegration of implant fixtures but also on maintaining the integrity of the connection of prosthetic superstructures to these fixtures. It was an objective of the present study to evaluate and compare cement bond strengths among rolled (R), cast (C) and metal-injection-molded (M) commercially pure titanium plates which were bonded with Panavia 21 (Kuraray) and Imperva (Shofu) cements. Two plates (15x5x1 mm) of each R, C, and M were lap-jointed (lap length: 5 mm). The joints were stored in 37 degrees C distilled water for 24 h, followed by tensile tests with an INSTRON system under 1 mm/min crosshead speed. It was found that the bond strength of R with Panavia 21 (PAN) was 5.31 (SD:1.5) MPa and 2.30 (0.83) MPa with Imperva (IMP) cement. These were improved by applying an alloy primer to 7.08 (1.31) MPa and 6.72 (1.63) MPa, respectively. Using PAN with primer application, C and M samples showed bond strengths of 7.99 (1.31) and 7.20 (2.50) MPa, while they were 5.83 (2.15) and 6.79 (2.09) MPa using IMP with primer. There was a significant difference (p<0.01) between PAN and IMP cements for C samples. Additionally, samples were pre-oxidized at 100 degrees C in air for 10 min. Bond strengths of PAN with the primer were 5.69 (2.25), 9.14 (1.28), and 5.60 (3.13) MPa for R, C, and M sample groups. If the cement with the primer was applied immediately after the polishing (instead of pre-oxidized surfaces), bond strengths were improved to 9.14 (1.78) for R, 9.29 (1.85) for C, and 9.36 (1.81) MPa for M sample group. At p<0.05 level, there was a significant difference between surface pre-condition of R and M, but no significance with C. PMID- 12883176 TI - Inhibition of Ni release from NiTi alloy by hydroxyapatite, alumina, and titanium sputtered coatings. AB - To alleviate the effects of Ni allergy from NiTi alloy implants, hydroxyapatite (Ca10(PO4)6(OH)2; HA), alumina (Al2O3), or titanium (Ti) was coated onto NiTi alloy plates to form 1-microm thick films using radio frequency magnetron sputtering. The coatings on the plates were characterized using XRD. After the plates had been immersed in physiological saline for periods of one, four, or eight weeks, the concentration of Ni ions released in each solution was detected using a microwave induced plasma mass spectrometer. After eight weeks, the concentration of Ni ions released from the non-coated, the Ti-coated, the HA coated, and the alumina-coated plates were 238, 19.7, 183, and 106 ppb, respectively. The bonding strength of the Ti film, the HA film, and the alumina film to the NiTi substrate were 3.8 +/- 1.2, 2.6 +/- 0.7, and 3.1 +/- 1.2 MPa, respectively. The non-coated, the HA-coated, the alumina-coated, and the Ti coated plates were implanted into the femurs of a dog for four weeks for histological observation. In case of the non-coated plates, connective tissue more than 300 microm thick was observed, whereas for the coated plates the thickness of the connective tissue was around 100 microm. PMID- 12883177 TI - Electrochemical study on microbiology-related corrosion of metallic dental materials. AB - Microbiology-related corrosion has been noted in industry for many years. It is widely recognized that microorganisms affect the corrosion of metal and alloys immersed in aqueous environments. Under similar conditions, the effect of bacteria in the oral environment on the corrosion of dental metallic materials remains unknown. The purpose of this study is to investigate the corrosion behavior of dental metallic materials in the presence of Streptococcus mutans and its growth byproducts. Samples were commercially pure titanium (CPT), Ti-6Al-4V (TAV), Ti-Ni (TN), Co-Cr-Mo alloy (CCM), 316L stainless steel (SSL), 17Cr-4Ni PH type stainless steel (PH), and Ni-Cr alloy (NC). Using Gamry corrosion test system, surfaces were exposed to (1) sterilized Ringer's solution as a control for (2), (2) S. mutans mixed with sterilized Ringer's solution; (3) sterilized tryptic soy broth as a control for (4), and (4) byproducts of S. mutans mixed with sterilized tryptic soy broth. Corrosion parameters (EOCP, ECORR, ICORR, etc.) were corrected for all tested samples. Averaged values of these parameters were statistically analyzed by t-test to identify significant differences. It was concluded that (1) S. mutans reduced the EOCP of CPT, TAV, TN, and SSL, and the byproducts of S. mutans reduced the EOCP of TAV, TN, SSL, and PH. (2) S. mutans increased the ICORR of PH, and byproducts of S. mutans increased the ICORR of all the samples. (3) S. mutans reduced the ECORR of CPT, TAV and TN, and the byproducts of S. mutans reduced the ECORR of TN, SSL, PH, and NC. (4) S. mutans increased the IPASS of CPT, and the byproducts of S. mutans increased the IPASS of CPT, PH, and NC. PMID- 12883178 TI - Design of a retractable intramedullary nail for the humerus. AB - This paper describes the design of a retractable intramedullary nail for the humerus that does not require inter-locking screws. The developed nail has a series of fins which open out from the casing to grip the medullary canal of the bone, thus securing it in position. Prototypes of the nail have been mechanically tested using static compression, dynamic compression and static torsion tests. During the compression tests no nails were found to fail. Statically a mean force of 809 N was withstood. During dynamic testing all the nails survived 1 million cycles, with a maximum applied load of 400 N. In torsion the two nails failed at 1.0 and 2.2 N m. Based on the mechanical testing, the retractable intramedullary nail would appear strong enough to withstand the expected loading conditions in the human body. PMID- 12883179 TI - The role of capsule composition and biologic responses in the function of transplanted microencapsulated islets of Langerhans. PMID- 12883180 TI - Role of ischemic preconditioning and the portosystemic shunt in the prevention of liver and lung damage after rat liver transplantation. AB - BACKGROUND: This study evaluates whether surgical strategies such as the portosystemic shunt and ischemic preconditioning can protect against hepatic and pulmonary injury associated with liver transplantation. METHODS: The effect of the portosystemic shunt, ischemic preconditioning, and both surgical procedures together were evaluated in rat liver transplantation. Alanine aminotransferase, hyaluronic acid levels in plasma, adenosine triphosphate and nucleotide levels in liver and edema, malondialdehyde levels, and myeloperoxidase activity were measured 24 hr posttransplantation. Plasmatic tumor necrosis factor (TNF) levels were measured as a possible proinflammatory factor responsible for hepatic and pulmonary damage associated with liver transplantation. RESULTS: Hepatocyte and cell endothelial damage were observed in liver grafts subjected to 8 hr of cold ischemia. This was associated with increased plasma TNF levels and lung inflammatory response. Portosystemic shunt application in the recipient protected endothelial cells but did not confer an effective protection from hepatocyte damage or reduce the increased plasma TNF levels and lung damage after liver transplantation. However, preconditioning of the donor liver conferred protection against both the endothelial cell and hepatocyte damage observed after liver transplantation. Preconditioning also attenuated the increased plasma TNF release and pulmonary damage. The combination of both surgical strategies resulted in levels of liver injury, TNF, and lung damage similar to those seen after liver transplantation. CONCLUSIONS: These findings indicate that ischemic preconditioning could be a preferred treatment to reduce hepatic and pulmonary damage associated with liver transplantation. However, this strategy may not be effective in several clinical situations requiring a portosystemic shunt. PMID- 12883181 TI - Subcutaneous transplantation of macroencapsulated porcine pancreatic endocrine cells normalizes hyperglycemia in diabetic mice. AB - BACKGROUND: The ultimate goal of islet transplantation is the unlimited availability of insulin-secreting cells to be transplanted in a simple procedure that requires no use of immunosuppressive drugs. Immunoisolation of xenogeneic pig islets for transplantation has great potential therapeutic benefits for treatment of diabetes. METHODS: Approximately 4 x 10(6) porcine pancreatic endocrine cells (PEC) isolated from 6-month-old pigs were macroencapsulated in agarose-poly(styrene sulfonic acid) mixed gel and implanted into a prevascularized subcutaneous site in streptozotocin-induced C57BL/6 diabetic mice. Animals receiving an equal number of free porcine PEC were used as controls. After transplantation, nonfasting blood glucose, body weight, intraperitoneal glucose tolerance test, and immunohistologic evaluations were processed. RESULTS: All 10 animals receiving the subcutaneous xenografts of the macroencapsulated porcine PEC normalized hyperglycemia within 5 days after transplantation, maintained the duration of normoglycemia for 24 to 76 days, and gradually gained weight. The subcutaneous xenografts of free porcine PEC could not reverse hyperglycemia. The recipient became hyperglycemic again when the implanted graft was retrieved at day 45 after transplantation. The glucose clearances were significantly ameliorated at day 21 and day 45 after transplantation when compared with those in diabetic mice. The immunohistochemical results revealed an inherent intact structure of the macroencapsulated porcine PEC and positive double-immunofluorescence staining for insulin and glucagon. CONCLUSIONS: Subcutaneous transplantation of macroencapsulated porcine PEC normalized hyperglycemia in diabetic mice. Our results identified a potential for a favorable development of subcutaneous transplantation of porcine PEC as a cure for diabetes. PMID- 12883182 TI - Donor lymphocyte infusion-mediated graft-versus-leukemia effects in mixed chimeras established with a nonmyeloablative conditioning regimen: extinction of graft-versus-leukemia effects after conversion to full donor chimerism. AB - BACKGROUND: We investigated an approach to separating graft-versus-lymphoma (GVL) effects from graft-versus-host disease (GVHD) in mice receiving a nonmyeloablative conditioning regimen allowing establishment of mixed hematopoietic chimerism. METHODS: We evaluated the ability of donor lymphocyte infusions (DLI) to mediate GVL effects without GVHD in mixed chimeras prepared with cyclophosphamide, anti-T-cell antibodies, and thymic irradiation. To examine the fate of GVH-reactive donor CD8+ T cells, we used the 2C T-cell receptor (TCR) transgenic mouse strain, which carries an Ld-specific transgenic TCR on the B6 background. RESULTS: Administration of DLI on day 35 post-BMT led to conversion from mixed to full donor chimerism and mediated a powerful GVL effect with complete protection (100% survival) against mortality induced by a host-type lymphoma (EL4) administered 7 days later (100% mortality in non-DLI controls; P<0.001). No GVHD occurred in DLI recipients. Rechallenging the surviving DLI recipients, which had converted to full chimerism, with the same tumor dose 17 weeks later led to rapid tumor mortality. Long-term DLI recipients had anti-host proliferative responses, but not CTL responses in vitro. When given as DLI together with wild-type spleen cells, marked expansion of GVH-reactive 2C CD8+ T cells was observed on day 10, followed by a marked decline in their numbers by week 10 post-DLI. CONCLUSIONS: Nonmyeloablative induction of mixed chimerism followed by administration of DLI can mediate powerful GVL effects. The late loss of DLI-mediated GVL effects may reflect the eventual loss of donor-derived GVH reactive CTL, which occurs in association with conversion to full donor chimerism. PMID- 12883183 TI - Amelioration of tacrolimus-induced nephrotoxicity in rats using juniper oil. AB - BACKGROUND: Calcineurin-inhibitor nephrotoxicity plays a role in the pathogenesis of chronic allograft nephropathy by causing renal ischemia mediated by vasoconstrictive metabolites of the prostanoid pathway. The purpose of our study was to evaluate whether altering the prostanoid profile using juniper oil (JO) would afford renoprotection in rats treated with tacrolimus. METHODS: Diets supplemented with biologic oils (no supplementation, JO, fish oil [FO], safflower oil [SO], and arachidonic acid [AA]) were fed to five groups of rats for 5 weeks; during the last 2 weeks, tacrolimus was administered to all groups except for a control group of animals. At week 5, urinary prostaglandin (PG)F(2-alpha) and inulin clearances were measured. The rat kidneys were harvested to determine the renal cell membrane composition for arachidonic, eicosatrienoic, and eicosapentaenoic acids. RESULTS: Both JO and FO completely reversed the decrease in inulin clearance seen with tacrolimus, the greatest effect being with JO (inulin clearance 15.1+/-3 vs. 6.0+/-1.1 ml/min in the nonsupplemented group; P<0.001); urinary PGF(2-alpha) excretion was also highest in the JO group (328+/ 23 pg/mL, P<0.001 vs. the nonsupplemented group). Fatty acid membrane analysis showed greatest incorporation of eicosapentaenoic and eicosatrienoic acids in the JO- (5.7+/-0.6% and 3.1+/-0.4%, respectively) and FO- (8.1+/-0.7% and 2.8+/-0.6%, respectively) treated animals. CONCLUSIONS: JO supplementation in tacrolimus treated rats was associated with incorporation of vasodilatory prostanoids in the renal-cell membrane and elevated urinary PGF(2-alpha) excretion, and the precipitous fall in inulin clearance induced by tacrolimus was completely prevented. Whether this benefit will translate into a reduction in chronic allograft nephropathy remains to be determined. However, our preliminary data point towards the need for human trials. PMID- 12883184 TI - A longitudinal study of children who received renal transplants 10-20 years ago. AB - BACKGROUND: We have previously reported on our 10-year experience of renal transplantation in children in the cyclosporine era, that is, from December 1981 until December 1991. In this paper, we report on the same children observed for another 10 years. METHODS: Of 53 children who received a renal transplant between 1981 and 1991, 47 survived and were observed for 10 to 20 years. Immunosuppression consisted of cyclosporine, prednisolone, and azathioprine. Yearly clinical examinations were performed. RESULTS: Overall, actual patient survival is 91%, 89%, and 89%, and actual graft survival 85%, 77%, and 66% at 1, 5, and 10 years, respectively. No patients have died during the last 10 years. Twenty-six grafts were lost over 20 years. Thirteen of those were lost during the present follow-up (10-20 years): 11 in chronic rejection and 2 because of development of renal cell carcinoma. No other malignancies were noted. Mean glomerular filtration rate decreased from 58+/-19 at 1 year (n=42) to 44+/-16 mL/min/1.73 m2 body surface area at 10 (n=33) years. Hypertension was treated in 46%, 40%, and 66% of the children at 1, 5, and 10 years, respectively; two of them showed left ventricular hypertrophy 10 years after transplant. Minor cataracts without visual disturbance were found in 45% of patients. All children except three with mental retardation are, or have been, attending normal day care or normal school. CONCLUSION: Social integration is good, and severe complications are scarce, even when renal transplantation occurred at a very young age. PMID- 12883185 TI - Sirolimus does not increase the risk for postoperative thromboembolic events among renal transplant recipients. AB - BACKGROUND: Deep venous thrombosis (DVT) tends to occur in greater frequency among cyclosporine (CsA)-treated renal-transplant recipients. Because administration of sirolimus may increase the whole-blood concentrations of CsA, we sought to assess the impact of the combination regimen on the incidence, predisposing factors, and consequences of postoperative DVT, transplant renal vein or artery thrombosis, and pulmonary embolus. METHODS: We retrospectively evaluated two cohorts of renal transplant recipients: CsA/prednisone (Pred)+/ azathioprine (n=136, group A) or sirolimus+CsA+Pred (n=354, group B) using Fisher's exact t and chi-square tests, as well as Kaplan-Meier analyses, odds ratios, and multiple logistic regression methods. RESULTS: The 7 of 136 (5.1%) incidence of thrombotic events in group A was similar to the 20 of 354 (5.6%) incidence in group B (P=0.513; NS) and occurred no more frequently ipsilateral to the transplant. Although the occurrence of an acute-rejection episode was not associated with the DVT diagnosis, all affected patients displayed elevated serum creatinine (Scr) values, which remained slightly higher than baseline following recovery (group A 1.63+/-1.22-1.95+/-0.93 mg/dL; group B 1.70+/-1.11-2.01+/-0.88 mg/dL). Renal biopsies failed to show evidence of intrarenal coagulopathy. No patient lost a graft as a complication of DVT, nor did these events produce other lasting adverse effects. Patients in the sirolimus group showed a strong correlation between the occurrence of DVT and the previous existence of an ipsilateral or contralateral lymphocele. CONCLUSION: Addition of sirolimus to a CsA+Pred regimen does not increase the incidence of postoperative thrombotic events among renal transplant recipients. PMID- 12883186 TI - Red blood cell alloantibodies and liver transplantation in Chinese patients. AB - BACKGROUND: Red blood cell (RBC) alloantibodies are present in up to 14% of white recipients of liver transplants and can cause severe delayed hemolysis. METHODS: A retrospective survey showed 17 cases (8.8%) of RBC alloantibodies in 192 consecutive Chinese recipients of liver transplants compared with a background hospital incidence of 3.7%. RESULTS: The spectrum of RBC alloantibodies in Chinese patients was different than in white patients, with no anti-D or anti-K antibodies but a significant incidence of anti-Mi (29%) antibodies. There was a significantly increased incidence of transfusions in RBC alloantibody positive cases. Delayed hemolysis also resulted in higher day-7 bilirubin levels. A total of 7 to 86 antigen-positive units were issued in five RBC alloantibody cases, including three early deaths. Seven cases in the RBC alloantibody negative group, but none in the positive group, were salvaged by regraft. CONCLUSIONS: Blood banks servicing transplant centers should be aware of ethnic patterns in RBC alloantibodies. Delayed hemolysis may jeopardize patient survival as the result of difficult postoperative stabilization, especially in cases requiring massive transfusion. PMID- 12883187 TI - Impact of hepatic rearterialization on reperfusion injury and outcome after mouse liver transplantation. AB - BACKGROUND: Nonarterialized mouse liver transplantation is a well-established model for immunologic studies on rejection and tolerance mechanisms. However, the importance of graft arterialization has-in contrast to rat liver transplantation not been thoroughly examined in the mouse model. The aim of the current study was to investigate the impact of arterial reconstruction on long-term graft survival, histologic alterations, ischemic liver damage, and early immunologic activation pathways. METHODS AND RESULTS: All recipients of arterialized (n=6) and nonarterialized (n=8) syngeneic liver grafts survived indefinitely. There were no differences in their histologic architecture, including no evidence of bile duct proliferation, periductal fibrosis, or alterations in serum transaminases, in long-term survivors from either group. Twenty-four hours after syngeneic liver transplantation, aspartate aminotransferase and alanine aminotransferase levels were increased to an equivalent extent in both groups, in agreement with early reperfusion injury and solitary traumatic injuries as assessed histologically (n=3 per group). Visualized by immunohistochemistry, intercellular adhesion molecule-1 expression was increased on sinusoidal and hepatic vein endothelium at both 1 and 100 days after transplantation, in both arterialized and nonarterialized grafts. Messenger RNA for interleukin-1, interferon-gamma, and tumor necrosis factor-alpha were measured by real-time polymerase chain reaction 24 hr after transplantation. No significant changes in the expression of cytokine mRNA levels were observed. CONCLUSIONS: Arterialization of mouse liver grafts does not appear to have a major impact on survival rate or the degree of immunologic activation. Therefore, the value of arterial reconstruction in mouse liver transplantation for experimental investigations is negligible. PMID- 12883188 TI - Iliac artery stenosis proximal to a kidney transplant: clinical findings, duplex sonographic criteria, treatment, and outcome. AB - BACKGROUND: Stenosis of the iliac segment proximal to the transplant renal artery (Prox-TRAS) is an uncommon cause of graft dysfunction and hypertension. We assessed the role of duplex sonography (DS) in regard to clinical and angiographic findings and followed the patients after percutaneous transluminal angioplasty (PTA), PTA stenting (PTAS), or surgery. METHODS: From January 1988 to August 2001, 97 of 1,064 kidney recipients underwent angiography for clinical or Doppler-sonographic suspicion of vascular problems. Kidney function, blood pressure, medication, and DS findings after renal transplantation (RTx) at the time of diagnosis of Prox-TRAS and after treatment were evaluated. RESULTS: Prox TRAS was diagnosed in 16 patients (1.5%) (49.6+/-6.9 years). Four patients demonstrated early presentation of Prox-TRAS 1 to 7 days after RTx (group A), leading to acute renal failure but without hypertension. In all patients, DS revealed pulsus parvus et tardus, low pulsatility index (PI) (<1.0), and a pathologic flow profile in the iliac artery proximal and distal to the graft. After treatment (surgery in two patients, PTA in one patient, PTAS in one patient), all patients developed good renal function (creatinine 1.7+/-0.9 mg/dL). PI increased from 0.9+/-0.1 to 1.2+/-0.1 (P=0.04), and flow profile within the iliac artery distal to the graft normalized. Late presentation (3-209 months after RTx) of Prox-TRAS was observed in 12 patients (group B), causing an increase of creatinine in 11 patients (two patients receiving dialysis treatments), impairment of blood pressure (141+/-15 and 80.7+/-7 to 160+/-18 and 85+/-7 mm Hg, P=0.009), and an increase in antihypertensive drugs (2.1+/-1.1 and 4.3+/-1, P=0.003) in all patients. The PI was decreased when compared with values early after RTx (1.6+/-0.4 to 1.2+/-0.3, P=0.007), and flow profile in the iliac artery was pathologic. All patients except one were managed by surgery (n=6), PTA (n=1), or PTAS (n=4). Creatinine (2.7+/-1.4 to 1.8+/-0.4 mg/dL, P=0.02) and blood pressure (160+/-18/85+/-7 mm Hg to 138+/-7/82+/-9, P=0.018) improved. Antihypertensive drugs were reduced to 2.8+/-0.8 (P=0.01). PI increased from 1.2+/-0.3 to 1.9+/-0.5 (P=0.01). Flow profile within the iliac artery distal to the graft anastomosis normalized. Kidney function, blood pressure, and PI remained unchanged during follow-up (82+/-69.9 months) in both groups. CONCLUSIONS: Prox-TRAS is rare. Because clinical symptoms are similar to those of transplant renal artery stenosis, DS is a valuable tool for diagnosis and follow up for this type of vascular lesion. Selective treatment with PTA, PTAS, or surgery improves kidney function and hypertension. PMID- 12883189 TI - Donors with central nervous system malignancies: are they truly safe? AB - BACKGROUND: In an era of organ shortage, the use of expanded or marginal donors has been attempted to increase transplantation rates and diminish waiting list mortality. One strategy is the use of organs from patients with a history of or active central nervous system (CNS) tumor. METHODS: Sixty-two recipients were identified as the recipients of organs from donors with a history of or active CNS malignancy. Patient demographics, donor tumor management, incidence of tumor transmission, and patient survival were examined. RESULTS: Of the organs recovered and transplanted from donors with astrocytoma, 14 were associated with at least one risk factor including high-grade tumor (n=4), prior surgery (n=5), radiation therapy (n=4), and systemic chemotherapy (n=4). One tumor transmission was identified at 20 months posttransplant with the patient expiring from metastatic disease. Twenty-six organs were transplanted from glioblastoma patients with 15 demonstrating risk factors including high-grade tumor (n=9) and prior surgery (n=10). Eight transmissions were identified with a range of 2 to 15 months posttransplant, with seven patients dying as the result of metastatic disease. Seven organs were used from donors with a medulloblastoma. Three transmissions were identified at a range of 5 to 7 months, all associated with ventriculoperitoneal shunts. Two medulloblastoma recipients died as the result of metastatic disease, whereas the third is alive with diffuse disease. The rate of donor tumor transmission, in the absence of risk factors, was 7%, whereas in the presence of one or more risk factor this rate dramatically rose to 53% (P<0.01). CONCLUSIONS: Organs from donors with CNS tumors can be used with a low risk of donor tumor transmission in the absence of the following risk factors: high-grade tumors, ventriculoperitoneal or ventriculoatrial shunts, prior craniotomy, and systemic chemotherapy. PMID- 12883190 TI - Use of steatotic graft in living-donor liver transplantation. AB - BACKGROUND: The degree of fatty infiltration in hepatic grafts is known to be an important risk factor for primary graft nonfunction in cadaveric liver transplantation. However, the effect of hepatic steatosis in living-donor liver transplantation (LDLT) has not yet been well defined. In this study, we evaluated the impact that the degree of hepatic graft steatosis has on the outcome of LDLT. METHODS: Sixty consecutive donors and recipients who underwent LDLT between October 1996 and August 2001 at Kyushu University Hospital were the subjects of this study. The pathologic findings of the prereperfusion biopsy of the graft were classified into the following three groups according to the degree of macrovesicular steatosis: None (n=23), 0% steatosis; Mild (n=23), 0% to 20% steatosis; and Moderate (n=6), 20% to 50% steatosis. Liver function tests including total bilirubin (at postoperative day [POD] 7), the peak alanine aminotransferase (ALT) and prothrombin time (at POD 3), and both patient and graft survival were compared among the groups. Furthermore, we also compared the donor parameters including the peak ALT and total bilirubin (at POD 3) and the operative time, blood loss, and length of hospital stay after surgery. RESULTS: The 1-year patient and graft survival were comparable among the groups. The peak ALT was significantly higher in the Moderate group (606+/-641 IU/L) than in the None (290+/-190 IU/L) and Mild (376+/-296 IU/L) groups. Total bilirubin (POD 7) and prothrombin time (POD 3) were comparable among the groups. The donor parameters were comparable among the groups except for the fact that the donor body weight of the Mild and Moderate groups were significantly heavier (P<0.0001) than that of the None group. CONCLUSIONS: In conclusion, the use of a fatty liver graft up to the moderate level can be justified in LDLT, even though ischemia reperfusion injury tends to be severe in such grafts. PMID- 12883191 TI - Analysis of the United Network for Organ Sharing database comparing renal allografts and patient survival in combined liver-kidney transplantation with the contralateral allografts in kidney alone or kidney-pancreas transplantation. AB - BACKGROUND: Combined liver-kidney transplantation (LKT) is the accepted treatment for patients with liver failure and irreversible renal insufficiency. Controversy exists as to whether simultaneous LKT with organs from the same donor confers immunologic and graft survival benefit to the kidney allograft. This study compares the outcomes of simultaneous LKT with the contralateral kidneys used for kidney alone transplantation (KAT) or combined pancreas-kidney transplantation (PKT) to understand the factors that account for the differences in survival. METHODS: From October 1987 to October 2001, LKTs with organs from 899 cadaver donors were reported to the United Network for Organ Sharing; 800 contralateral kidneys from these donors were used in 628 KAT and 172 PKT recipients. These 800 paired control patients were the basis of this analysis. RESULTS: Graft and patient survival rates were lower among LKT recipients compared with KAT (P<0.001) and PKT recipients (P<0.001), because of a higher patient mortality rate during the first 3 months posttransplant. Among human leukocyte antigen mismatched transplants, LKT recipients demonstrated the highest 1-year rejection free survival rate (LKT 70%, KAT 61%, and PKT 57% ) (P=0.005 vs. KAT, P=0.005 vs. PKT). There was a lower incidence of renal graft loss resulting from chronic rejection among LKT recipients (LKT 2% vs. KAT 8% vs. PKT 6%, P<0.0001). CONCLUSIONS: Patients undergoing LKT exhibit a higher rate of mortality during the first year posttransplant compared with patients undergoing KAT and KPT. Analysis of the data indicates an allograft-enhancing effect of liver transplantation on the renal allograft. PMID- 12883192 TI - Multislice computed tomography angiography in pediatric liver transplantation. AB - BACKGROUND: Preoperative delineation of any vascular anomalies offers planning for possible alteration of surgical procedures, especially in pediatric recipients undergoing living-related liver transplantation. PURPOSE: We assess the efficacy of three-dimensional (3D) multislice computed tomography (CT) angiography in the hope of replacing conventional angiography as the pretransplant evaluation of the hepatic vascular system for potential recipients of liver transplantation. METHODS: 3D CT angiography was performed in 38 children with biliary atresia. Conventional angiography was also performed in the first 15 patients. Twelve patients underwent living-related liver transplantation. The findings on 3D CT angiography were compared with conventional angiography and operative findings. RESULTS: 3D CT angiography was successfully performed in 37 pediatric patients. All findings of 3D CT angiography on hepatic artery, portal vein, and inferior vena cava paralleled those of catheter angiography and operative findings. Four patients were unsuitable to receive living grafts because of pathologic insults of the hepatic artery (one patient) and the portal vein (three patients). Three patients were advised to undergo a venous graft for portal anastomoses. Eight patients demonstrated portosystemic shunts that may require closure. CONCLUSION: 3D CT angiography proves to be a better tool in the demonstration of the vascular system and identification of pathologic insults in pediatric patients. It is superior to conventional angiography because it is less invasive, more convenient, and more efficient in providing thorough preoperative information that would have a major impact on patient selection and surgical planning. PMID- 12883193 TI - Relationship between postoperative erythromycin breath test and early morbidity in liver transplant recipients. AB - BACKGROUND: Interindividual variability in dosage requirements of the calcineurin inhibitor immunosuppressive agents cyclosporine and tacrolimus after liver transplantation may result from differences in the CYP3A activity of the liver graft. Early postoperative erythromycin breath test (ERMBT) is an in vivo measure of graft CYP3A activity. This study evaluates the usefulness of an early postoperative ERMBT in predicting early morbidity in liver transplant recipients. METHODS: In 26 liver transplant recipients, ERMBT was performed within 2 hr after transplantation. Main end points were the occurrence of cyclosporine and tacrolimus nephrotoxicity, episodes of early graft rejection, early graft function, and graft survival. RESULTS: Cyclosporine and tacrolimus nephrotoxicity were associated with low postoperative ERMBT values (mean 0.63%+/-0.25% 14C/hr vs. 1.35%+/-0.84% 14C/hr, P=0.02). No significant association between early graft rejection and ERMBT values was demonstrated. There was a significant inverse correlation between postoperative ERMBT values and the time to normalization of international normalized ratio as a measure of early graft function (r=-0.78, P<0.001). Graft loss was associated with low postoperative ERMBT values (0.21%+/ 0.15% 14C/hr vs. 1.09%+/-0.72% 14C/hr, P=0.002). CONCLUSION: An early postoperative ERMBT may be useful in predicting the development of cyclosporine and tacrolimus nephrotoxicity, severe graft dysfunction, or even graft loss in liver transplant recipients when calcineurin inhibitors are administered according to protocols. Whether ERMBT results may be used to individualize dosage of calcineurin inhibitors needs to be explored. PMID- 12883194 TI - Long-term improvement in renal function with sirolimus after early cyclosporine withdrawal in renal transplant recipients: 2-year results of the Rapamune Maintenance Regimen Study. AB - INTRODUCTION: The purpose of this study was to evaluate early cyclosporine (CsA) withdrawal from a sirolimus (SRL)-CsA-steroid (ST) regimen. METHODS: Within 48 hr after transplantation, 525 primary (90%) or secondary (10%) renal allograft recipients with cadaveric (89%) or living (11%) donors received 2 mg of SRL (troughs >5 ng/mL; immunoassay), CsA, and ST. Those eligible (430) were randomly assigned (1:1) at 3 months +/- 2 weeks to remain on triple-drug therapy (SRL-CsA ST group) or to have CsA withdrawn and SRL trough concentrations targeted to 20 to 30 ng/mL (SRL-ST group) until month 12, and 15 to 25 ng/mL thereafter. RESULTS: At 24 months, there were no statistically significant differences in patient survival (94.0% vs. 95.3%), graft survival (91.2% vs. 93.5%), acute rejection after randomization (5.1% vs. 9.8%) or discontinuations (34% vs. 33%) for SRL-CsA-ST versus SRL-ST, respectively. Serum creatinine level was significantly better in patients who had CsA withdrawn (167 vs. 128 micromol/L, P<0.001), as was the slope of 1/creatinine. Similarly, systolic blood pressure was lower in patients who had CsA withdrawn (141 vs. 134 mm Hg, P<0.001). High density lipoprotein cholesterol was significantly higher in the SRL-ST group, whereas total cholesterol, low-density lipoprotein cholesterol, and triglyceride levels were not significantly different. Hypertension, creatinine increase, abnormal kidney function, toxic nephropathy, edema, hyperuricemia, cataracts, Herpes zoster, and malignancy were reported significantly more often in patients continuing CsA. Thrombocytopenia, hypokalemia, abnormal liver function tests, abnormal wound healing, ileus, and pneumonia were reported significantly more frequently with SRL-ST. CONCLUSION: Data at 2 years confirm that early CsA withdrawal followed by an SRL-ST maintenance regimen results in long-term improvement in both renal function and blood pressure, without increased risk of graft loss or late acute rejection. PMID- 12883195 TI - Kidney and liver transplantation in human immunodeficiency virus-infected patients: a pilot safety and efficacy study. AB - BACKGROUND: Human immunodeficiency virus (HIV)-infected patients have historically been excluded from consideration for transplantation out of concern for the effects of immunosuppression on the progression of HIV disease. Improvements in HIV-related morbidity and mortality with the use of highly active antiretroviral therapy (HAART) have prompted a reevaluation of transplantation as a treatment option for HIV-infected patients with end-stage kidney and liver disease. METHODS: Eligible patients met standard transplant criteria. They had undetectable plasma HIV-1 RNA levels (viral load) for 3 months (kidney) or were predicted to achieve viral load suppression posttransplantation if unable to tolerate HAART (liver); a CD4+ T-cell count of more than 200 cells/microL (kidney) or more than 100 cells/microL (liver) for 6 months; and no history of opportunistic infections and neoplasm. Standard immunosuppression included prednisone, mycophenolate mofetil (CellCept, Roche Pharmaceuticals, Basel, Switzerland), and cyclosporine (Neoral, Novartis, East Hanover, NJ). RESULTS: Fourteen patients received transplants (10 kidney transplants, mean follow-up 480 days; four liver transplants, mean follow-up 380 days). All of the kidney transplant recipients (100%) are alive and with functioning grafts, and three of four liver transplant patients (75%) are alive and well with functioning grafts (all liver transplant patients with normal liver function tests). The one death occurred 445 days posttransplantation in a liver recipient coinfected with hepatitis C virus, who died as the result of its rapid reoccurrence. Rejection occurred in 5 of 10 kidney transplant recipients but did not occur in any of the four liver transplant recipients. HIV viral loads have remained undetectable in all patients maintained with HAART. CD4 counts have remained stable in patients not treated for rejection. Patients receiving protease inhibitors require 25% of the dose of cyclosporine compared with patients receiving nonnucleoside reverse transcriptase inhibitors. CONCLUSIONS: There has been no evidence of significant HIV progression and no adverse effect of HIV on allograft function. Rejection is a concern in kidney transplant recipients, as is the possible poor outcome in hepatitis C virus-coinfected liver transplant recipients. Preliminary data are encouraging and indicate that transplantation should be a treatment option for individuals with well-controlled HIV disease. PMID- 12883196 TI - Dyslipidemia in renal transplant recipients treated with a sirolimus and cyclosporine-based immunosuppressive regimen: incidence, risk factors, progression, and prognosis. AB - BACKGROUND: This retrospective study compared the incidence, severity, and predisposing factors for dyslipidemia among renal transplant patients treated for up to 6 years with a cyclosporine +/- prednisone-based concentration-controlled regimen without (n=118) or with (n=280) ascending exposures to sirolimus. METHODS: The diagnosis of dyslipidemia was established when the serum cholesterol value (CHO) was more than 240 mg/dL or serum triglycerides (TG) were more than 200 mg/dL. Generalized estimating equations and mixed-modeling procedures were used for statistical analyses. RESULTS: Hypercholesterolemia was observed in 46% to 80% and hypertriglyceridemia in 43% to 78% of sirolimus-treated patients during the first 6 posttransplantation months. The mean peak serum lipid levels among patients in the sirolimus group (CHO=285.5 mg/dL; TG=322.4 mg/dL) were significantly higher than those in the nonsirolimus group (CHO=250.2 mg/dL and TG=267.6 mg/dL; both P<0.01). The lipid values, which were persistently elevated during the first posttransplantation year, decreased slowly thereafter but remained significantly higher than the pretransplantation levels beyond 4 years after transplantation. The two forms of hyperlipidemia tended to occur in parallel (Pearson's coefficient of correlation, r=0.5, P<0.001), showing a positive predictive value of 0.67 and a negative predictive value of 0.65. However, there was no significant difference in the incidence of cardiovascular events within 4 years after transplantation among patients treated with versus without sirolimus. CONCLUSION: The dyslipidemia associated with sirolimus therapy, albeit persistent, does not seem to represent a major risk factor for the early emergence of cardiovascular complications. PMID- 12883197 TI - Effects of adenoviral up-regulation of bcl-2 on oxidative stress and graft coronary artery disease in rat heart transplants. AB - BACKGROUND: Bcl-2 has been shown to have antioxidant properties. Early oxidative stress is an important antigen-independent factor that contributes to the development of graft coronary artery disease (GCAD). We hypothesized that adenoviral up-regulation of bcl-2 would decrease early oxidative stress and inhibit GCAD after heart transplantation. METHODS: PVG rat hearts were treated with adenovirus carrying the human bcl-2 gene (AdvBcl-2) or null adenovirus (AdvNull) then transplanted into the abdomens of PVG recipients. After 4 days of reperfusion to allow adenoviral gene expression, grafts were retransplanted into ACI rat recipients and reperfused for 4 or 8 hours or 90 days (cyclosporine A 7.5 mg/kg on postoperative day [POD] 0-9). Production of tumor necrosis factor (TNF) alpha after 4 hours and oxidized glutathione (GSSG) after 8 hours indicated development of oxidative stress. 90-day allografts were assessed for GCAD by way of computerized morphometry. RESULTS: Over-expression of bcl-2 at the time of allograft reperfusion was confirmed by Western blotting. Whereas AdvNull-treated hearts demonstrated elevated TNF-alpha levels after 4 hours and increased GSSG after 8 hours of reperfusion, AdvBcl-2-treated hearts were no different from nontransplanted hearts. AdvBcl-2 treatment also resulted in decreased luminal narrowing and intima-to-media ratio at POD 90. CONCLUSIONS: Bcl-2 over-expression interrupts the development of oxidative stress in reperfused rat-heart allografts. Early up-regulation of bcl-2 also decreases GCAD, indicating the importance of early oxidative stress and the role that bcl-2 may play in the long term function of heart transplants. PMID- 12883198 TI - Class II transactivator promoter activity is suppressed through regulation by a trophoblast noncoding RNA. AB - BACKGROUND: Trophoblasts lack expression of all classic major histocompatibility complex (MHC) antigens. Determination of the mechanism involved could provide insight into selective gene suppression and allograft tolerance. Suppression of class II expression in trophoblasts is secondary to dominant negative trans acting factors that suppress class II transactivator (CIITA) transcription. We recently described a trophoblast-derived noncoding RNA (TncRNA) that suppresses class II expression. We examined the effects of TncRNA on the CIITA promoter, CIITA, and MHC class II expression. METHODS: HeLa clones stably transfected with TncRNA were analyzed for MHC class II and CIITA expression by fluorescence activated cell sorting, Northern blots, and quantitative polymerase chain reaction. Activity and functional dissection of CIITA promoter IV (pIV) was assessed by transient co-transfection of promoter-reporter constructs. Methylation of pIV was assessed by Southern blots, fluorescence-activated cell sorting, and quantitative polymerase chain reaction. RESULTS: TncRNA suppressed interferon-gamma-induced human leukocyte antigen-DR and CIITA expression in HeLa cells. The mechanism involves inhibition of CIITA pIV through a defined inhibitory domain on the promoter. The mechanism does not involve methylation of the promoter. CONCLUSIONS: A novel method of CIITA suppression is described where a noncoding RNA selectively mediates the suppression of CIITA pIV possibly by complementary RNA-DNA binding to an inhibitory domain on the promoter. Selective suppression of MHC class II could have important implications in allograft tolerance and in developing class II-deficient cells or tissues for the purpose of transplantation or drug delivery systems. PMID- 12883199 TI - Detection of donor-specific anti-HLA antibodies with flow cytometry in eluates and sera from renal transplant recipients with chronic allograft nephropathy. AB - BACKGROUND: Chronic allograft nephropathy (CAN), which remains the main cause of graft loss after kidney transplantation, is still poorly understood. Because anti HLA antibodies may be involved in the pathogenesis of CAN, this study was performed to look for donor-specific antibodies (DSA) fixed onto renal transplants with CAN. METHODS: DSA were identified after elution with flow cytometric assay and/or flow cytometric crossmatches in 20 transplants removed after irreversible graft failure caused by CAN and in control samples from 2 transplants with relapsing glomerulopathy, 2 transplants lost after vascular thrombosis, and 4 normal kidneys. The results were compared with those obtained in the serum samples 1 year after grafting, at the time of transplantectomy, and within 2 months after transplantectomy. RESULTS: IgG anti-class I, anti-class II, or both DSA were identified in 70.6% of eluates versus 73.6% of posttransplantectomy serum samples (NS), 42.1% of 1-year postgrafting serum samples (P<0.05), and 31.6% of serum samples at the time of transplantectomy (P<0.05). Our data show a good correlation between the target of anti-HLA antibodies found in both eluates and posttransplantectomy serum samples, but the precise specificity of anti-HLA antibodies is more often assigned in posttransplantectomy serum samples than in eluates. This problem needs further evaluation. CONCLUSION: This study shows that testing for anti-HLA DSA in eluates from removed kidney transplants using flow cytometry can be achieved and is highly efficient. It already suggests that both anti-class I and anti-class II HLA antibodies can be involved in CAN. Further studies are now needed to evaluate the possibility of identifying such antibodies in the eluates of transplant biopsy specimens from recipients experiencing CAN. PMID- 12883200 TI - Alteration in CD45RBhi/CD45RBlo T-cell ratio following CD45RB monoclonal-antibody therapy occurs by selective deletion of CD45RBhi effector cells. AB - Tolerance induction by CD45RB monoclonal antibody (mAb) in murine allograft models is associated with an alteration in the CD45RBlo/CD45RBhi T-cell ratio in favor of CD45RBlo T cells, which can function as regulatory cells and promote tolerance. It has been proposed that inversion of the CD45RBhi/CD45RBlo normal T cell ratio by mAb can occur by down-regulation of CD45RB surface molecules expressed by T cells. Because CD45RB mAb infusion can lead to a reduction in peripheral T cells, we tested whether other mechanisms might participate in the inversion of the CD45RBhi/CD45RBlo ratio, including apoptosis of CD45RBhi cells. We report that CD45RB mAb led to rapid elimination of both CD4+ and CD8+ T cells in vitro. Importantly, CD45RB mAb selectively eliminated CD45RBhi T cells without affecting the viability of CD45RBlo T cells. Furthermore, the death of T cells occurred with a reduction in mitochondrial transmembrane potential and DNA fragmentation but with little evidence of nuclear condensation and cell shrinkage typically found with cells undergoing apoptosis. We propose that CD45RB mAb therapy may promote a dominant regulatory T-cell population that has the capacity to inhibit rejection by the selective elimination of CD45RBhi effector T cells. This occurs by a process that does not involve the classic morphologic features of apoptosis. Strategies that facilitate an inversion of the CD45RBhi/CD45RBlo T cell subset ratio may improve the efficacy of CD45RB mAb, and therapeutic measures that prevent deletion of CD45RBhi T cells may need to be avoided to achieve tolerance clinically. PMID- 12883201 TI - Effects of rewarming on nuclear factor-kappaB and interleukin 8 expression in cold-preserved alveolar epithelial cells. AB - BACKGROUND: Nuclear factor-kappaB (NF-kappaB) and interleukin (IL)-8 play important roles in the pathophysiology of acute lung injury after lung transplantation. Because alveolar epithelium is one of the most important sites at which IL-8 production takes place after reperfusion of donor lungs, we examined the effects of cold/rewarming on NF-kappaB and IL-8 expression in alveolar epithelial cells. METHODS: A549 cells were preserved at 4 degrees C for 5 hr and then rewarmed for up to 20 hr. NF-kappaB was analyzed by electrophoretic mobility shift assay. IL-8 mRNA expression was examined by reverse transcription polymerase chain reaction. IL-8 concentration in the cell culture medium after rewarming was measured by enzyme-linked immunosorbent assay. RESULTS: NF-kappaB was increased in the nuclear extracts as early as 30 min after rewarming. There was a marked increase in the IL-8 mRNA expression at 1 and 3 hr after rewarming. IL-8 concentration in the cell culture medium was progressively increased during 20 hr following rewarming. The cell culture medium inhibited apoptosis of neutrophils significantly. The cold/rewarming-induced IL-8 production was reduced to approximately 50% by introducing an antisense oligonucleotide for the p65 subunit of NF-kappaB and by treatment with N-acetyl-leucinyl-leucinyl-norleucinal and pyrrolidine dithiocarbamate. The effect of dexamethasone treatment was dose dependent (reduced to approximately 30% at 10-5 M dexamethasone). CONCLUSIONS: Our results indicate that rewarming of cold-preserved alveolar epithelial cells itself may be an important initiator of the inflammatory cascades, including NF kappaB activation and IL-8 release. Inhibition of NF-kappaB would be worth trying to control unnecessary IL-8 production and the inflammatory response in the donor lungs. PMID- 12883202 TI - Breakdown of peripheral T-cell tolerance by chronic interleukin-15 elevation. AB - Thymic deletion purges the repertoire of most developing T cells with the potential for overt self-reactivity, but some self-specific cells do emerge into the peripheral pool. Under most conditions, these potentially autoaggressive cells remain in a quiescent state. However, in some circumstances, they become activated and acquire effector function, leading to immune disease. It is thus important to clarify the mechanism(s) responsible for determining the balance between such inappropriate T-cell activation and the normal state of peripheral tolerance. In this article, we show that chronic elevation of interleukin-15 levels interferes with the tolerant state of CD8+ T cells through a process that involves activation of nonlymphoid antigen-presenting cells by CD4+asialo-GM1+ (ASGM1) or both CD4+ASGM1- and CD4-ASGM1+ cells. These findings suggest a potential role for dysregulated interleukin-15 production in promoting tolerance breakdown. This new information may be of potential use in improving tumor vaccines to self-antigens and in ameliorating autoimmune or graft-versus-host disease. PMID- 12883203 TI - Time course of proteinuria after living-donor kidney transplantation. AB - BACKGROUND: After cadaveric kidney transplantation, preservation-reperfusion damage results in glomerular and tubular proteinuria. There are no data on the time course of proteinuria after living-donor (LD) transplantation. METHODS: In 10 patients receiving a kidney graft from an LD, the excretion of high molecular weight proteins (albumin, transferrin, and immunoglobulin G) and low molecular weight proteins (beta2-microglobulin and alpha1-microglobulin) was measured at various time points during the first 5 days after transplantation. RESULTS: Immediately after restoration of the circulation, we observed a massive nonselective high molecular weight proteinuria, indicative of glomerular damage. This proteinuria rapidly decreased to slightly elevated values beyond 24 hr after transplantation. Low molecular weight proteinuria, reflecting tubular damage, was also prominent and remained grossly abnormal even at day 5. CONCLUSION: After LD transplantation, preservation-reperfusion injury causes massive proteinuria during the first 24 hr. Thereafter proteinuria rarely exceeds 1 g per day. PMID- 12883204 TI - Increased levels of syndecan-1 in serum during acute graft-versus-host disease. AB - Acute graft-versus-host disease (GVHD) may affect several organs. Syndecan-1 is a heparan sulfate proteoglycan that can be shed from the surface of most epithelial cells (skin, liver, and gut among others), which are target organs for GVHD. Syndecan-1 was measured in serum samples from 60 patients after allogeneic stem cell transplantation and was related to the degree of GVHD. Syndecan-1 levels increased in patients who developed acute GVHD but not in patients without GVHD. The difference between groups was significant 3 to 10 weeks after transplantation. The peak level of syndecan-1 in serum correlated with the degree of acute GVHD (r=0.46, P<0.001). Combined, the peak levels of syndecan-1 and soluble interleukin-2 receptor detected patients with acute GVHD (sensitivity 82%, specificity 89%). This study shows that syndecan-1 levels are increased during acute GVHD. Syndecan-1 may be a marker for acute GVHD, especially if combined with determination of soluble interleukin-2 receptor. PMID- 12883205 TI - Higher surgical wound complication rates with sirolimus immunosuppression after kidney transplantation: a matched-pair pilot study. AB - Sirolimus, a potent new immunosuppressant, has been anecdotally associated with surgical wound complications. We studied postoperative surgical wound complications in 15 kidney recipients receiving sirolimus, prednisone, and tacrolimus or cyclosporine (study group) compared with 15 recipients receiving tacrolimus, prednisone, and mycophenolate mofetil who were pair-matched for surgical wound complication risk factors. Surgical wound complications were defined as any complication related to the surgical transplant wound requiring reintervention. Fifty-three percent of the study group and 7% of the control group experienced more than one surgical wound complication (P=0.014), and the relaparotomy incidence was 33% and 7%, respectively. Four graft losses have occurred since the beginning of the study: one chronic rejection and two deaths with function in the study group, and one death with function in the control group. At 1 year, graft survival for study recipients compared with control recipients was 87% and 93%, respectively; patient survival was 93% in both groups. Recipients receiving sirolimus demonstrated a significantly higher surgical wound complication rate, but graft and patient survival were not affected. Peritransplant immunosuppression with sirolimus and steroids warrants careful consideration, particularly in recipients with surgical complication risk factors. PMID- 12883206 TI - Late-onset cytomegalovirus reactivation in critically ill renal transplant patients. AB - BACKGROUND: Cytomegalovirus (CMV) reactivation occurs frequently in the first months after renal transplantation. However, reports concerning long-term kidney transplant recipients are rare and have always pertained to symptomatic CMV disease. METHODS: We report four cases of late-onset asymptomatic CMV reactivation in critically ill renal transplant patients who suffered from severe bacterial infections and in whom CMV antigenemia was observed. RESULTS AND CONCLUSION: CMV reactivation in these patients might indicate an additional disturbance in the patients' immune defenses at the time of critical illness, possibly even necessitating a temporary reduction in immunosuppressive therapy. Prospective, controlled trials are needed to define the role of CMV antigenemia in critically ill patients, including the role of antiviral therapy for asymptomatic reactivations. PMID- 12883207 TI - Elimination of false-negative hepatitis C virus RNA results by removal of inhibitors in cadaver-organ donor blood specimens. AB - Detection of viral nucleic acids in blood samples from cadavers is often difficult because of inhibition of the reverse transcriptase (RT) or polymerase chain reaction (PCR) steps by substances present in the samples. A robust method for the extraction and detection of hepatitis C virus (HCV) RNA from cadaver blood samples by polymerase chain reaction RT-PCR has been developed on the basis of the Qiagen QIAamp DNA mini kit extraction system (Basel, Switzerland). Twenty of 36 samples tested were positive for HCV RNA. Six of the 16 HCV-antibody- and RNA-negative samples contained inhibitors that were successfully removed by pretreatment of samples with the Qiagen AX matrix before extraction. PMID- 12883208 TI - Progressive multifocal leukoencephalopathy after allogeneic stem cell transplantation and posttransplantation rituximab. PMID- 12883209 TI - Protective role of bone marrow cells in controlling alloreactivity. PMID- 12883212 TI - Difficulty of achieving long-term graft survival of MHC-disparate composite graft using CTLA4IG. PMID- 12883213 TI - Response to "Proteinuria in normal cadaver kidney donors". PMID- 12883214 TI - Successful liver transplantation from an eclamptic donor complicated by the HELLP syndrome. PMID- 12883215 TI - Use of mycophenolate mofetil in immunosuppressive protocols in elderly renal transplant recipients. PMID- 12883216 TI - Vascular closure staples for experimental organ transplantation. PMID- 12883217 TI - Assessment of renal function in the failing graft: a difficult task. PMID- 12883218 TI - C2 monitoring: a reliable tool in pediatric renal transplant recipients. PMID- 12883219 TI - Noninvasive diagnosis of BK virus nephritis by measurement of messenger RNA for BK VP1 virus in urine. PMID- 12883220 TI - Cardiac hypertrophy in liver transplant recipients: tacrolimus, cyclosporine or both? PMID- 12883222 TI - A case of tacrolimus (FK506)-induced pancreatitis and fatality 2 years postcadaveric renal transplant. PMID- 12883223 TI - Changing trends in the epidemiology of pediatric lead exposure: interrelationship of blood lead and ZPP concentrations and a comparison to the US population. AB - OBJECTIVES: To determine blood lead and zinc protoporphyrin (ZPP) concentrations in a pediatric population, confirm their interrelationship at low blood lead concentrations, and assess changing trends through comparison of these data with those found in a similar population 10 years earlier and to US national values. STUDY DESIGN AND METHODS: The study was conducted in a large pediatric hospital in the Washington DC area (CNMC) on patient whole blood specimens (n = 4908) (0 17 years) accrued from January 2001 to June 2002. Pediatric blood lead concentrations were determined by atomic absorption spectrophotometry, and ZPP by hematofluorometry. The data were analyzed using a computer adaptation of the Hoffmann approach. RESULTS AND CONCLUSIONS: Blood lead level (BLL) means ranged between 2.2 and 3.3 micro g/dL, and the median BLL was 3 micro g/dL throughout. Mean ZPP concentrations ranged between 21.1 and 26.6 micro g/dL and the median concentrations between 21 and 27 micro g/dL. In comparison to data obtained from a similar pediatric population at CNMC between 1991 and 1992, pediatric BLLs have significantly declined in the Washington DC area. The current data are also compared with data obtained from the National Health and Nutrition Examination Survey (NHANES III) of the US population. The interrelationship between ZPP and BLLs is examined. PMID- 12883224 TI - Neonatal hair analysis contribution to establishing a gestational drug exposure profile and predicting a withdrawal syndrome. AB - Recently, interest in hair analysis in such fields as drug abuse, driving, or for clinical purposes (determination of drug-exposed neonates especially) has grown because of the highly sensitive method of detection (GC-MS) that can now be applied. Neonates born to drug-addicted mothers can suffer from neonatal withdrawal syndrome (NWS), which requires morphine treatment in its severe forms. To assess and measure toxicologic factors predicting the appearance and the severity of this syndrome, matrices such as urine, meconium, and hair are necessary. Cannabinoids, opiates, cocaine (and its metabolites), and methadone in particular were determined in the various matrices collected in 17 mother/neonate pairs. An immunologic screening method was used, and quantification was achieved with GC-MS. In spite of some bias (color, length, race) that might hinder an accurate interpretation, the results of hair analysis makes it possible to confirm a fetal drug exposure and to reinforce the diagnosis of the NWS observed, particularly when results obtained in other matrices are negative. Hair analysis contributes to our ability to predict a NWS. PMID- 12883225 TI - Determination of epirubicin and its metabolite epirubicinol in saliva and plasma by HPLC. AB - We present a high-performance liquid chromatography (HPLC) method suitable for the analysis of epirubicin and its metabolite epirubicinol in saliva and plasma. Preparation of saliva and plasma samples was performed by extraction of analytes with a chloroform:2-propanol mixture (6:1, vol/vol) and evaporation of the organic phase to dryness under vacuum at a temperature of approximately 45 degrees C. The chromatographic analysis was carried out by reversed-phase isocratic elution of the anthracyclines with a Chromsep stainless steel HPLC column (150 x 4.6 mm I.D.) filled with Nucleosil 100 S C(18) material, particle size 5 micro m. The detection was accomplished by spectrofluorimetry at excitation and emission wavelengths of 474 and 551 nm, respectively. The anthracyclines eluted within 10 min of injection, and the method appeared to be specific. The method is linear over a concentration range of 5 to 1000 micro g/L for epirubicin and 2 to 400 micro g/L for epirubicinol (r > 0.99) in both saliva and plasma. The recoveries from saliva and plasma of epirubicin, epirubicinol, and the internal standard doxorubicin were 88.9 and 69.0%, 87.6 and 77.3%, and 80 and 67.9%, respectively. The lower limit of quantification was 5 micro g/L for epirubicin and 2 micro g/L for epirubicinol. The method proved to be precise and accurate, as the within-day and between-day coefficients of variation were less than 10%. Overall results indicate that our method is suitable for the bioanalysis of epirubicin and epirubicinol in saliva as well as plasma. PMID- 12883226 TI - An automated HPLC method to determine intracellular vincristine concentrations in mononuclear cells of children with acute lymphoblastic leukemia. AB - A method to determine intracellular vincristine concentrations in vivo in leukemic cells of patients is useful to investigate mechanisms of vincristine resistance. We developed a high-performance liquid chromatographic (HPLC) method to measure vincristine concentrations in human mononuclear cells (MNC). This method, with on-line column solid-phase extraction and electrochemical detection, previously developed for determination of vincristine concentrations in human plasma, was validated for determination of intracellular vincristine concentrations over a range of 1.17 to 50.8 micro g/L with a lower limit of quantitation (LOQ) of 1.17 micro g/L. Linearity of the relationship between concentration and response was characterized by a slope (sd) of 0.0683 (0.0008), an intercept (sd) of 0.0004 micro g/L (0.0085), and a regression coefficient of 0.997 (P = 0.05). Maximum bias and within-day and between-day coefficients of variation (CV) were 14.2%, 4.8%, and 5.7%, respectively. The HPLC detector response did not interfere with the vincristine peaks of five independent MNC sources. The method was used successfully to measure intracellular vincristine concentrations in 8 of 35 bone marrow MNC samples of children newly diagnosed with acute lymphoblastic leukemia (ALL), 3 days after the first injection of 1.5 mg/m2 vincristine. Vincristine concentrations were in the range of 4.0 to 26.4 micro g/L, which was 5 to 20 times the bone marrow plasma concentration. The described HPLC method is accurate and precise and is suitable for detection of intracellular vincristine concentrations in MNC of children with ALL. The results confirm that in vivo vincristine accumulates in bone marrow MNC of patients. PMID- 12883227 TI - Long-term pharmacokinetic study of the novel combination of tacrolimus and sirolimus in de novo renal allograft recipients. AB - It was recently shown in two randomized studies that combining sirolimus (rapamycin) and tacrolimus is very efficient in renal transplantation. However, little is known about the long-term pharmacokinetics of this combination. We performed simultaneous AUC measurements (area under the concentration curves) of sirolimus and tacrolimus at 1, 3, and 12 months posttransplantation in nine de novo recipients treated with this drug combination to characterize the evolution of the pharmacokinetics of both drugs and to investigate possible interactions between the two compounds. Patients were treated with a standard-dose tacrolimus or with a reduced-dose tacrolimus in combination with sirolimus and corticosteroids. This long-term pharmacokinetic study has shown that when sirolimus is combined with tacrolimus, dose changes of sirolimus are reflected by pharmacokinetic exposure parameters. Patients taking a low dose of sirolimus in combination with a standard dose tacrolimus might require sirolimus dose increments over time to maintain constant exposure to sirolimus. Further prospective dose-controlled studies are necessary to investigate a possible effect of a standard-dose tacrolimus on long-term sirolimus bioavailability and/or metabolism. Dose reductions of tacrolimus in both study groups were reflected by concordant decreasing pharmacokinetic exposure parameters, which illustrates the common clinical practice of reducing the dose of calcineurin inhibitor as time elapses after transplantation. PMID- 12883228 TI - Measurement of beta-methyldigoxin level in serum from patients by enzyme immunoassay using novel specific antiserum with a phenyl boric acid column. AB - The authors compared serum beta-methyldigoxin (MDx) levels in digitalized patients by enzyme immunoassay (EIA) using anti-MDx 3'-hemisuccinate BSA antiserum (antiserum-I) with commercial antidigoxin antiserum (antiserum-II). The usefulness of a phenyl boric acid (PBA) column for pretreatment of the serum samples was also investigated. The assay using antiserum-I demonstrated good accuracy and precision in the concentration range of 0.5 to 5 ng/mL. When the specificities of antiserum-I and antiserum-II were assessed by cross-reactivity studies with various related compounds, antiserum-I was much more specific for MDx antiserum-II. Using a phenyl boric acid (PBA) column, MDx, and digoxigenin, which exhibits a negligible cross-reactivity, were separated from serum, including MDx and its metabolites. The recovery tests of MDx using antiserum-I with a PBA column in human serum were satisfactory and no interference of metabolites of MDx was observed. Mean MDx concentrations in serum samples (n = 30) from digitalized patients by EIA using antiserum-I with PBA column, antiserum I, and antiserum-II were 1.06, 1.30, and 1.74 ng/mL, respectively. These results indicate that our EIA system using antiserum-I with a PBA column for pretreatment of serum samples is useful to more precisely measure the unchanged type of MDx in patients. PMID- 12883229 TI - Serum concentrations and effects of gabapentin and vigabatrin: observations from a dose titration study. AB - To explore possible concentration-effect relationships, gabapentin (GBP) and vigabatrin (VGB) serum concentrations were obtained from patients participating in an add-on dose-titration trial comparing GBP and VGB in partial epilepsy. Patients randomized to GBP started on 1800 mg/d and could have their dosage increased stepwise to 2400 and 3600 mg/d if seizures persisted. Those randomised to VGB started on 1000 mg/d, and the dose could be increased to 2000 and 4000 mg/d. Blood samples were obtained at steady state, at a nonstandardized time, from 27 patients randomized to GBP and from 36 randomized to VGB. Serum samples were analyzed using high-performance liquid chromatography. The treatment effect was expressed as percentage reduction in number of seizures from baseline. In addition, patients were classified as responders (>50% reduction in number of seizures from baseline) or nonresponders. There was no significant correlation between serum concentrations of GBP and seizure reduction at the lowest dosage, 1800 mg/d (r = -0.02, P = 0.94, Spearman-rank), nor between VGB serum levels and seizure reduction at 1000 mg/d of VGB (r = -0.14, P = 0.44). The serum GBP concentrations among responders to GBP 1800 mg/d were 26 +/- 12 micro mol/L (mean +/- SD), which was not different from serum concentrations in nonresponders, 28+/ 13 micro mol/L. Nor was there a difference between serum concentrations of responders and nonresponders to VGB 1000 mg/d (32 +/- 23 and 44 +/- 36 micro mol/L, respectively). Hence, with the present study design we were unable to identify specific target ranges of GBP and VGB serum concentrations. PMID- 12883230 TI - Effects of the CYP 2D6 genotype and cigarette smoking on the steady-state plasma concentrations of fluvoxamine and its major metabolite fluvoxamino acid in Japanese depressed patients. AB - The effects of the cytochrome P450 (CYP) 2D6 genotype and cigarette smoking on the steady-state plasma concentrations (C(ss)) of fluvoxamine (FLV) and its demethylated metabolite fluvoxamino acid (FLA) were studied in 49 Japanese depressed patients receiving FLV 200 mg/d. The C(ss) of FLV and FLA were measured by HPLC, and the wild-type allele (*1) and two mutated alleles causing absent (*5) or decreased (*10) CYP 2D6 activity were identified by PCR methods. The patients were divided into three genotype groups by the number of mutated alleles: 12 cases with no (*1/*1), 27 cases with one (*1/*5 and *1/*10), and 10 cases with two (*5/*10 and *10/*10) mutated alleles. The means +/- SD of the C(ss) of FLV and FLA and the FLA/FLV ratio of all patients were 169.1 +/- 147.5 ng/mL, 83.9 +/- 52.7 ng/mL, and 0.71 +/- 0.50, respectively. The C(ss) of FLV and FLA were not significantly different among the three genotype groups. However, the FLA/FLV ratio was significantly lower in the patients with one (P < 0.05) and two (P < 0.01) mutated alleles than in those with no mutated allele. There was no significant difference between nonsmokers (n = 34) and smokers (n = 15) in these values. In the stepwise multiple regression, the C(ss) of FLA (P < 0.05) and FLA/FLV ratio (P < 0.001) showed significant negative correlations with the number of mutated alleles, and the FLA/FLV ratio was significantly (P < 0.05) lower in women than in men. The present study suggests that the CYP 2D6 genotype and cigarette smoking have no major impact on the C(ss) of FLV and FLA, though CYP 2D6 is involved in the demethylation of FLV. PMID- 12883231 TI - Monitoring clozapine: are fingerprick blood and plasma clozapine levels equivalent to arm venipuncture blood and plasma levels? AB - The objective of this pilot study was to determine whether fingerprick blood and plasma clozapine levels were equivalent to arm venipuncture blood and plasma levels for the purpose of therapeutic monitoring. A convenient sample of 10 outpatients from the Elgin Program of Assertive Community Treatment Team (PACT) participated in the study. Blood samples were obtained simultaneously from both the arm and finger in patients at steady state to measure clozapine levels. Each site provided a blood and plasma clozapine level, and they were compared. Clozapine levels from arm and finger sites were found to be equivalent in both blood and plasma. Although plasma clozapine levels were consistently greater than those in whole blood by a mean value of 27%, the plasma therapeutic threshold level (350-400 micro g/L) was considered an adequate target for monitoring. A fingerprick blood sample of 50 micro L was sufficient to measure clozapine levels accurately at steady state. We therefore concluded that fingerprick blood testing is as effective as the traditional arm venipuncture method in obtaining accurate clozapine levels. This procedure may provide certain benefits for the seriously mentally ill. PMID- 12883232 TI - Effects of itraconazole on the plasma kinetics of quazepam and its two active metabolites after a single oral dose of the drug. AB - The effects of itraconazole, a potent inhibitor of cytochrome P450 (CYP) 3A4, on the plasma kinetics of quazepam and its two active metabolites after a single oral dose of the drug were studied. Ten healthy male volunteers received itraconazole 100 mg/d or placebo for 14 days in a double-blind randomized crossover manner, and on the fourth day of the treatment they received a single oral 20-mg dose of quazepam. Blood samplings and evaluation of psychomotor function by the Digit Symbol Substitution Test and Stanford Sleepiness Scale were conducted up to 240 h after quazepam dosing. Itraconazole treatment did not change the plasma kinetics of quazepam but significantly decreased the peak plasma concentration and area under the plasma concentration-time curve of 2 oxoquazepam and N-desalkyl-2-oxoquazepam. Itraconazole treatment did not affect either of the psychomotor function parameters. The present study thus suggests that CYP 3A4 is partly involved in the metabolism of quazepam. PMID- 12883233 TI - A new turbidometric digoxin immunoassay on the ADVIA 1650 analyzer is free from interference by spironolactone, potassium canrenoate, and their common metabolite canrenone. AB - Spironolactone and potassium canrenoate (aldosterone antagonist diuretics) are often used with digoxin in clinical practice. It has been well documented in the literature that spironolactone, potassium canrenoate, and their common metabolite canrenone cross-react with the fluorescence polarization immunoassay (FPIA) for digoxin and falsely elevate measured serum digoxin concentrations. Recently a new turbidometric assay for digoxin became commercially available from Bayer Diagnostic for application on the ADVIA 1650 Chemistry analyzer. We studied the potential interference of these compounds in this new digoxin assay. Aliquots of drug-free serum were supplemented with therapeutic and above-therapeutic concentrations of spironolactone, canrenone, and potassium canrenoate, and apparent digoxin concentrations were measured. We observed apparent digoxin concentrations with the FPIA digoxin assay as expected but observed no apparent digoxin levels with the new turbidometric immunoassay. When serum pools prepared from patients receiving digoxin were supplemented with these compounds in concentrations expected in serum in patients receiving these medications, we observed falsely elevated digoxin levels with the FPIA digoxin assay, but no statistically significant change was observed with the new turbidometric assay. We conclude that the new turbidometric assay for digoxin is free from interference by spironolactone, potassium canrenoate, and their common metabolite canrenone. PMID- 12883234 TI - Itraconazole-tacrolimus drug interaction. PMID- 12883235 TI - "Lack of clinically significant interference by spironolactone with the Axsym Digoxin II assay" only applies to low-dose therapy with spironolactone. PMID- 12883236 TI - The immunohistochemical diagnosis of mesothelioma: a comparative study of epithelioid mesothelioma and lung adenocarcinoma. AB - A large number of immunohistochemical markers that can facilitate the distinction between epithelioid pleural mesotheliomas and pulmonary peripheral adenocarcinomas have recently become available. The aim of this study is to compare the value of these new markers with others that are already commonly used for this purpose and to determine which are, at present, the best for discriminating between these malignancies. Sixty epithelioid mesotheliomas and 50 lung adenocarcinomas were investigated for expression of the following markers: calretinin, cytokeratin 5/6, WT1, thrombomodulin, mesothelin, CD44S, HBME-1, N cadherin, E-cadherin, MOC-31, thyroid transcription factor-1 (TTF-1), BG-8 (Lewisy), carcinoembryonic antigen (CEA), Ber-EP4, B72.3 (TAG-72), leu-M1 (CD15), CA19-9, epithelial membrane antigen (EMA), and vimentin. All (100%) of the mesotheliomas reacted for calretinin, cytokeratin 5/6, and mesothelin, 93% for WT1, 93% for EMA, 85% for HBME-1, 77% for thrombomodulin; 73% for CD44S, 73% for N-cadherin, 55% for vimentin, 40% for E-cadherin, 18% for Ber-EP4, 8% for MOC-31, 7% for BG-8, and none for CEA, B72.3, leu-M1, TTF-1, or CA19-9. Of the adenocarcinomas, 100% were positive for MOC-31, Ber-EP4, and EMA, 96% for BG-8, 88% for CEA, 88% for E-cadherin, 84% for B72.3, 74% for TTF-1, 72% for leu-M1, 68% for HBME-1, 48% for CD44S, 48% for CA19-9, 38% for mesothelin, 38% for vimentin, 30% for N-cadherin, 14% for thrombomodulin, 8% for calretinin, 2% for cytokeratin 5/6, and none for WT1. After analyzing the results, it is concluded that calretinin, cytokeratin 5/6, and WT1 are the best positive markers for differentiating epithelioid malignant mesothelioma from pulmonary adenocarcinoma. The best discriminators among the antibodies considered to be negative markers for mesothelioma are CEA, MOC-31, Ber-EP4, BG-8, and B72.3. A panel of four markers (two positive and two negative) selected based upon availability and which ones yield good staining results in a given laboratory is recommended. Because of their specificity and sensitivity for mesotheliomas, the best combination appears to be calretinin and cytokeratin 5/6 (or WT1) for the positive markers and CEA and MOC-31 (or B72.3, Ber-EP4, or BG-8) for the negative markers. An extensive and detailed review of the literature is also provided. PMID- 12883237 TI - Microglandular adenosis with transition into adenoid cystic carcinoma of the breast. AB - Microglandular adenosis (MGA) is a well-recognized, if rare and incompletely characterized, entity in which carcinoma is rarely thought to develop. We report 17 cases in which patterns of adenoid cystic carcinoma (ACC) coexisted with MGA. Immunocharacterization with beta-catenin, E-cadherin, cytokeratins (AE1/AE3), epithelial membrane antigen, S-100 protein, smooth muscle actin, and vimentin was also performed. Most cases had areas of invasive ACC characterized by its defining dual-lumen types. Some cases of ACC appeared to have expanded glands intermingled within the MGA, whereas in other cases ACC formed a transition with the characteristic small, gland-like spaces of MGA. MGA and "atypical MGA" stained irregularly and similarly to that seen in myoepithelium with the three markers of myoepithelial cells in breast: S-100 protein, smooth muscle actin, and vimentin. These markers were also positive in the more solid elements of the ACC. Our study suggests that ACC may develop in a background of and in continuity with MGA. Altered myoepithelial cells appear to be the major neoplastic element in both ACC and "atypical MGA." "Atypical MGA" with transition to ACC may show histologic patterns and an immunohistochemical profile similar to that of ACC. These lesions might be best interpreted as ACC in situ. Both MGA and ACC of the breast grow in an expansile and diffusely infiltrative pattern without having significant metastatic capacity. Their unusual interaction with the surrounding stroma may play a role in this benign biologic behavior. PMID- 12883238 TI - Primary cutaneous marginal zone B-cell lymphoma: a molecular and clinicopathologic study of 24 asian cases. AB - Cutaneous marginal zone B-cell lymphoma is a recently proposed entity and constitutes a cutaneous counterpart of extranodal marginal zone B-cell lymphoma of mucosa-associated lymphoid tissue (MALT). Borrelia burgdorferi infection has been suggested as a possible causative agent in European cutaneous cases of marginal zone B-cell lymphoma, whereas API2-MALT1 fusion and BCL10 mutation are highly associated with MALT lymphoma. Aberrant nuclear BCL10 expression may be closely correlated with API2-MALT1 fusion in gastric and pulmonary MALT lymphomas. We examined 24 Asian cases of cutaneous marginal zone B-cell lymphoma for B. burgdorferi involvement, API2-MALT1 fusion, BCL10 cellular expression, and BCL10 mutation. Neither Borrelia DNA nor API2-MALT1 fusion transcript was detected. Nuclear BCL10 expression was evident in tumor cells of 11 of 24 cases, although BCL10 mutation was found in one case only. Clinicopathologically, nuclear BCL10 was more frequently expressed in macroscopically nodular lesions than in plaques or papules (p = 0.0031). These data suggest that 1) B. burgdorferi infection may not play an important role in developing cutaneous marginal zone B-cell lymphoma in Asian cases, 2) neither API2-MALT1 fusion nor BCL10 mutation is closely associated with the pathogenesis, 3) aberrant nuclear BCL10 may frequently be expressed in the absence of these genetic abnormalities, and 4) nuclear BCL10 expression may be clinically important because it was observed in locally aggressive tumors. PMID- 12883239 TI - Mucin-rich variant of salivary duct carcinoma: a clinicopathologic and immunohistochemical study of four cases. AB - Salivary duct carcinoma is a relatively uncommon aggressive neoplasm, typically found in the parotid glands of older men. The histologic appearance is that of an in situ and invasive high-grade adenocarcinoma, and it closely resembles ductal carcinoma of the breast. Several variants of the latter are very well known, but only papillary, sarcomatoid, and low-grade subtypes have so far been reported in salivary duct carcinoma. This study describes the clinicopathologic and immunohistochemical findings in four examples of an additional previously undescribed variant, rich in mucin. Each tumor showed areas of typical salivary duct carcinoma, but in addition there were lakes of epithelial mucin-containing malignant cells, i.e., mucinous (colloid) carcinoma. All four tumors expressed androgen receptors, cytokeratins, epithelial membrane antigen, gross cystic disease fluid protein-15, and carcinoembryonic antigen, but S-100 protein, other myoepithelial markers, and estrogen and progesterone receptors were negative. The mucin antigen profile showed positivity for MUC2, MUC5B, and MUC6 in all cases but only rare staining with MUC5AC and MUC7. Strong immunohistochemical overexpression of HER2/neu was demonstrated in one tumor, together with amplification by fluorescence in situ hybridization; another case was weakly positive with just one antiserum, but the remaining two tumors were completely negative. Small quantities of mucin have often been described in salivary duct carcinoma but not large extracellular mucinous lakes, which though prominent in the present series, were not as extensive as in mucinous adenocarcinoma. The relatively poor clinical outcome of the patients in our study mirrored that seen in usual-type salivary duct carcinoma and emphasizes the importance of differentiating mucin-rich salivary duct carcinoma from pure mucinous (colloid) adenocarcinoma, a tumor not fully defined, but possibly with a better prognosis. PMID- 12883240 TI - Endometrial endometrioid adenocarcinoma with a deceptive pattern of spread to the uterine cervix: a manifestation of stage IIb endometrial carcinoma liable to be misinterpreted as an independent carcinoma or a benign lesion. AB - The prognosis of endometrial endometrioid adenocarcinoma is determined in part by stage; endocervical stromal involvement (stage IIB) imparts a worsened prognosis. We describe a deceptive pattern of stage IIB disease that mimics a primary endocervical glandular proliferation and may lead to understaging of endometrial endometrioid adenocarcinoma. Fifteen cases of endometrial endometrioid adenocarcinoma with a peculiar pattern of cervical involvement were identified from our consultation files. All cases were referred in consultation because of doubt about the nature of the cervical process and its relation to the corpus tumor; in a few instances, the cervical proliferation was considered possibly benign and in one case was misinterpreted as mesonephric hyperplasia. The patients ranged from 49 to 84 years in age (mean age 64.9 years). There was usually a grossly evident endometrial tumor. The cervix was unremarkable grossly in at least 11 patients. The cervical tumors were composed of variably shaped, often tubular glands with little or no stromal response and mainly invaded as widely spaced glands that often appeared deceptively benign. In 14 cases luminal secretions, mainly eosinophilic, were identified, often leading to consideration of a mesonephric lesion. Ten of the endometrial tumors were grade 1, four grade 2, and one grade 3. One was noninvasive, nine superficially invasive, and five deeply invasive. In four cases myoinvasion had, at least in part, a diffusely infiltrative pattern. The tumors in the cervix showed no in situ component and no definite surface involvement. Continuity with the corpus tumor could be demonstrated in 12 cases. Ten of the cervical tumors invaded more deeply than the endometrial tumor, four invaded to a similar depth, and only one was more superficial than its endometrial counterpart. The cervical and corpus tumors had a similar immunoprofile in nine cases: all were vimentin positive, eight estrogen positive and one negative, four carcinoembryonic antigen negative, and five with focal apical or rare cytoplasmic staining. This immunoprofile in conjunction with routine morphologic similarity between the two tumors and the usual documented continuity between them indicate that the cervical process represents spread from the endometrial endometrioid adenocarcinoma. It is important for both therapeutic and prognostic reasons that the cervical abnormality is not misinterpreted as a benign or malignant primary endocervical glandular process. PMID- 12883241 TI - Appendiceal mucinous neoplasms: a clinicopathologic analysis of 107 cases. AB - The classification of appendiceal mucinous tumors is controversial and terminology used for them inconsistent, particularly when they lack overtly malignant features but are associated with extra-appendiceal spread. We reviewed 107 appendiceal mucinous neoplasms and classified them as low-grade appendiceal mucinous neoplasm (LAMN) (n = 88), mucinous adenocarcinomas (MACAs) (n = 16), or discordant (n = 3) based on architectural and cytologic features. LAMNs were characterized by a villous or flat proliferation of mucinous epithelium with low grade atypia. Thirty-nine tumors were confined to the appendix, but 49 had extra appendiceal tumor spread, including 39 with peritoneal tumor characterized by mucin pools harboring low-grade mucinous epithelium, usually dissecting in a hyalinized stroma. Eight of the 16 MACAs lacked destructive invasion of the appendiceal wall and eight showed an infiltrative pattern of invasion. Extra appendiceal tumor spread was present in 12 MACAs (four peritoneum, seven peritoneum and ovaries; one ovaries only). In MACAs with an infiltrative pattern, peritoneal tumor consisted of glands and single cells in a desmoplastic stroma. The peritoneal tumor in the remaining cases consisted of mucin pools that contained mucinous epithelium with high-grade atypia and, in some cases, increased cellularity compared with that seen in peritoneal spread in cases of LAMN. Three cases were classified as discordant because the appendiceal tumors were LAMNs but the peritoneal tumors were high-grade. Follow-up was available for 49 LAMNs, 15 MACAs, and 2 discordant cases. None of the patients with LAMNs confined to the appendix experienced recurrence (median follow-up 6 years). LAMNs with extra-appendiceal spread were associated with 3-, 5-, and 10-year survival rates of 100%, 86%, and 45%, respectively. Patients with MACA had 3- and 5-year survival rates of 90% and 44%, respectively (p = 0.04). The bulk of peritoneal disease correlated with prognosis among patients with MACA (p = 0.04) and, to a lesser degree, among patients with LAMNs (p = 0.07). We conclude that: 1) appendiceal mucinous neoplasms can be classified as either low-grade mucinous neoplasms or mucinous adenocarcinoma based on architectural and cytologic features; 2) tumors that can be confidently placed in the low-grade group (which requires rigorous pathologic evaluation of the appendix) and are confined to the appendix are clinically benign in our experience to date; 3) low-grade tumors confined to the appendix are morphologically identical to those with extra appendiceal spread (except for the usual identification of breach of the wall in the latter cases) and the same designation is appropriate for the appendiceal neoplasia in each situation; 4) the long-term outlook for patients with low-grade tumors and peritoneal spread is guarded with just over half dying of disease after 10 years; 5) appendiceal mucinous tumors with destructive invasion of the appendiceal wall, complex epithelial proliferations, or high-grade nuclear atypia generally pursue an aggressive clinical course and should be classified as mucinous adenocarcinomas; 6) peritoneal tumor can be classified as involvement by LAMN or MACA, and this distinction is of prognostic significance; 7) bulky peritoneal tumor worsens prognosis; and 8) LAMNs associated with high-grade peritoneal tumor behave as adenocarcinoma. PMID- 12883242 TI - Waldenstrom macroglobulinemia involving extramedullary sites: morphologic and immunophenotypic findings in 44 patients. AB - Waldenstrom macroglobulinemia (WM) is a clinicopathologic syndrome in which a B cell neoplasm involving the bone marrow, usually lymphoplasmacytic lymphoma (LPL), is associated with immunoglobulin M paraprotein in the serum. Extramedullary involvement occurs in a subset of patients and is infrequently examined histologically. The files of M.D. Anderson Cancer Center were searched for patients with WM who underwent biopsy of one or more extramedullary sites during the course of disease. Each biopsy specimen was classified using the criteria of the World Health Organization classification. The study group consisted of 44 patients (26 men and 18 women), with a total of 51 specimens obtained from lymph nodes (n = 36), soft tissue (n = 4), spleen (n = 3), skin (n = 2), lung (n = 2), tonsils (n = 1), colon (n = 1), liver (n = 1), and gallbladder (n = 1). Lymphoplasmacytic lymphoma was the most common histologic type, in 40 (78%) samples. This category was morphologically heterogeneous and was further subclassified as lymphoplasmacytic (n = 21), lymphoplasmacytoid (n = 18), and polymorphous (n = 1). Four of these LPL cases morphologically resembled marginal zone B-cell lymphoma. Four additional samples were involved by diffuse large B-cell lymphoma, probably transformed from LPL. Three more samples were involved by LPL with unusual features: two were CD5-positive and one was a composite tumor with classical Hodgkin's disease. Other categories of lymphoma in this group of patients with WM included small lymphocytic lymphoma/chronic lymphocytic leukemia (n = 2), mantle cell lymphoma (n = 1), and follicular lymphoma (n = 1). Waldenstrom macroglobulinemia is most commonly associated with LPL but can rarely occur with other types of B-cell lymphoma. Lymphoplasmacytic lymphoma in patients with WM is morphologically heterogeneous and can be indistinguishable from marginal zone B-cell lymphoma. CD5+ B-cell lymphomas with features otherwise typical of LPL are rare, and we think these tumors are part of the spectrum of LPL. PMID- 12883243 TI - Breast tumor resembling the tall cell variant of papillary thyroid carcinoma: report of 5 cases. AB - Five cases of a hitherto undescribed breast tumor having histologic features similar to those of the tall cell variant papillary thyroid carcinoma are described. They were composed of columnar mitochondrion-rich to oxyphilic cells arranged in nests, papillae, and follicle-like structures. In addition, the neoplastic cells showed numerous nuclear grooves and, in two cases, nuclear pseudo-inclusions. None of the patients had previous concomitant or subsequent evidence of a thyroid tumor. Immunohistochemistry further excluded a metastasis from the thyroid in the four cases tested, as they were consistently thyroglobulin and thyroid transcription factor 1 negative. PMID- 12883244 TI - Idiopathic chronic pancreatitis with periductal lymphoplasmacytic infiltration: clinicopathologic features of 35 cases. AB - To clarify clinicopathologic features of idiopathic chronic pancreatitis with lymphoplasmacytic infiltration, we carried out a study of 35 cases. There were two histologic groups, which we have designated lymphoplasmacytic sclerosing pancreatitis and idiopathic duct-centric chronic pancreatitis. Lymphoplasmacytic sclerosing pancreatitis (22 cases) was a fibrosing process with diffuse lymphoplasmacytic infiltrates involving pancreatic lobules and ducts, adipose tissue, blood vessels, and common bile duct. Obliterative phlebitis was found in every case except for one. The histologic features were similar to other idiopathic fibrosclerosing disorders, and one patient also had retroperitoneal fibrosis. Affected patients tended to be elderly men. Idiopathic duct-centric chronic pancreatitis (13 cases) was characterized by inflammatory infiltrates (including neutrophils) that were denser in the lobules than in interlobular fibrotic areas. Neutrophils were also prominent in the ducts, and destruction of the duct epithelium was commonly seen. Patient ages were more broadly distributed than in lymphoplasmacytic sclerosing pancreatitis. Two patients had inflammatory bowel disease. We conclude that idiopathic chronic pancreatitis with lymphoplasmacytic infiltration, sometimes called autoimmune pancreatitis, consists of at least two different processes. One of these, lymphoplasmacytic sclerosing pancreatitis, is a histologically unique lesion and could be a pancreatic manifestation of idiopathic fibrosclerosing disorders. PMID- 12883245 TI - Alpha-methylacyl-CoA racemase: a variably sensitive immunohistochemical marker for the diagnosis of small prostate cancer foci on needle biopsy. AB - Expression of the alpha-methylacyl-CoA racemase (AMACR) gene has recently been demonstrated by several groups to be markedly elevated in prostate cancer cells with little expression in benign prostate tissue and has been suggested as a molecular marker of prostate cancer on needle biopsy. There is scant data, however, as to the sensitivity and specificity of AMACR in the diagnosis of small foci of cancer on needle biopsy. A total of 209 needle biopsies of the prostate with small foci (<5% of a core) of prostatic adenocarcinoma were identified. A total of 175 cases were received in consultation by one of the authors (140 from a single institution and 35 from different outside institutions) and 34 cases were from our hospital file. Immunohistochemistry for high molecular weight cytokeratin and p63 was performed in all cases to confirm the diagnosis of cancer. Only AMACR staining that was significantly stronger than that of background benign glands was considered positive; 88% of all cases of prostate cancer were positive for AMACR. The sensitivity varied among the different groups: 100% for the in house cases, 87.1% for the cases from a single institution, and 80% for cases from different outside institutions. The mean percentage of stained glands in positive cases was 95.9%, with 150 (71.8%) cases showing 100% of the glands positive and 25 (12.0%) cases showing no staining. Because negative staining for basal cell markers, especially in a small focus of atypical glands, is not necessarily diagnostic of prostate cancer, positive staining for AMACR can increase the level of confidence in establishing a definitive malignant diagnosis. However, the sensitivity of AMACR staining may vary in specimens from different pathology laboratories, possibly related to differences in fixation and processing. It is important to optimize the staining technique for each laboratory and recognize that some small cancers on needle biopsy may be AMACR negative. PMID- 12883246 TI - The clinical significance of focal active colitis in pediatric patients. AB - The clinical significance of focal neutrophilic infiltrates in crypt epithelium in colorectal biopsies or focal active colitis has been studied in adult populations, but little is known about this entity in children. The incidence of Crohn's disease in adult patients presenting with focal active colitis has varied between 0% and 13% in previous studies, whereas the incidence of infectious-type colitis has been reported to be nearly 50%. We reviewed 31 cases of focal active colitis diagnosed in pediatric patients without a history of inflammatory bowel disease between 1989 and 2000. Pathologic variables studied included number and location of inflamed crypts and distribution and character of lamina propria inflammation. Clinical follow-up was obtained from patient charts. Two patients were lost to follow-up. Follow-up on the remaining 29 patients ranged from 4 months to 7 years with a mean of 4.2 years. Eight patients (27.6%) developed Crohn's disease. Nine patients (31%) appeared to have acute infectious-type colitis, one with C. difficile. Eight patients (27.6%) had focal active colitis, which did not correlate with their symptoms or ultimate clinical diagnosis. These were termed idiopathic focal active colitis. Two patients were found to have allergic colitis, one had ulcerative colitis, and one had Hirschsprung's disease. Pediatric patients with focal active colitis have a much higher incidence of Crohn's disease than adults with same entity. Hence, it is important to document the presence of focal active colitis in pediatric patients. PMID- 12883247 TI - An assessment of the expanded classification of congenital cystic adenomatoid malformations and their relationship to malignant transformation. AB - The purpose of this study was to review cases of congenital cystic adenomatoid malformations (CCAMs) arising in children and adults, in order to assess the recently expanded classification system for these lesions and their association with malignant transformation. Of 28 CCAMs, there were 16 type 1, 4 type 2, and 8 type 4 lesions, 12 of which presented in adults. Five of 16 type 1 CCAMs were accompanied by microscopic foci of bronchioloalveolar carcinoma; two others showed focal mucous cell hyperplasia. In two further cases, foci of nonmucinous atypical adenomatous hyperplasia were identified in the adjacent lung parenchyma. The bronchioloalveolar carcinomas showed less cytologic atypia, proliferative activity (Ki-67), and p53 expression than a comparative group of bronchioloalveolar carcinomas arising de novo, but this was not statistically significant (p = 0.15). Neither bronchioloalveolar carcinomas nor hyperplasia was identified in type 2 or type 4 CCAMs. Four of the eight type 4 CCAMs showed focal stromal hypercellularity, and one case subsequently developed a pleuropulmonary blastoma. We conclude that classification according to the current system is of clinical value. Bronchioloalveolar carcinomas arise in association with type 1 CCAMs, but recurrence following resection is exceptional. Type 4 CCAMs show histologic overlap with grade 1 pleuropulmonary blastomas, and distinction between these entities may not be possible on histology alone. However, stromal cellularity in a type 4 CCAM should raise the possibility of blastomatous transformation. PMID- 12883248 TI - Histopathology of interval (delayed) appendectomy specimens: strong association with granulomatous and xanthogranulomatous appendicitis. AB - Patients who present with a ruptured acute appendicitis are often treated with antibiotic therapy and drainage followed by a delayed or interval appendectomy. We noticed interval appendectomy specimens with granulomatous inflammation and postulated that interval appendectomy may lead to granulomatous appendicitis. To test this hypothesis, we reviewed the histopathology of all interval appendectomy specimens within a 4-year period and compared them with a control group of patients who had acute appendicitis and underwent routine acute appendectomy. All slides were randomized and reviewed blindly to assess the inflammatory patterns, with special attention given to the presence of granulomas and other Crohn-like features. Twenty-two cases of interval appendectomy were found. The interval between symptom onset and appendectomy ranged from 30 to 95 days with a mean of 58 days, whereas all 44 control patients had surgery within 72 hours of symptoms onset. Thirteen (59.1%) of the 22 interval appendectomy cases contained granulomas compared with only 3 of 44 controls (P < 0.0001). Eight (36.4%) of the interval appendectomy cases had xanthogranulomatous inflammation compared with none in the acute appendicitis group (P < 0.0001). A Crohn-like appearance was seen in 11 (50.0%) of the interval appendectomy cases and 1 of the controls (P < 0.0001). Follow-up data were available in 8 of 11 cases with Crohn-like features; none developed Crohn disease during an average follow-up period of 23 months. Delayed or interval appendectomy specimens often have a characteristic inflammatory pattern that includes granulomas, xanthogranulomatous inflammation, mural fibrosis/thickening, and transmural chronic inflammation. Without the appropriate clinical history, these changes may be misinterpreted as Crohn disease. PMID- 12883249 TI - Inhibin alpha distinguishes hemangioblastoma from clear cell renal cell carcinoma. AB - Inhibin alpha subunit (inhibin A) expression in hemangioblastomas has not been previously reported in the literature. We analyzed the expression of inhibin A in 25 hemangioblastomas from 22 patients. Eleven cases were from 8 patients with von Hippel-Lindau disease, and these tumors were multicentric and/or recurrent. The remaining 14 cases from 14 patients were sporadic. The male-to-female ratio was 8:3, and the age at presentation ranged from 19 to 78 years (mean 35 years; median 45 years). Eighteen tumors were located in the cerebellum/posterior fossa, 1 in the medulla, 1 in the occipital lobe, and 5 in the spinal cord. Four metastatic renal cell carcinomas in brain, 10 renal cell carcinomas from 8 patients with von Hippel-Lindau disease, and 5 sporadic clear cell renal cell carcinomas were also included. Two patients with von Hippel-Lindau disease had both renal cell carcinoma and hemangioblastoma. The stromal cells of all 25 cases of hemangioblastoma expressed inhibin A. Strong, moderate, and weak cytoplasmic immunoreactivity was noted in 17, 5, and 3 cases, respectively. In contrast, none of the 19 renal cell carcinomas, primary as well as metastatic, expressed inhibin A. There was no difference in the inhibin A staining pattern between the sporadic hemangioblastoma and those associated with VHL. These findings demonstrate inhibin A to be a useful marker in distinguishing hemangioblastoma from metastatic clear cell renal cell carcinoma. While the diagnostic importance is evident, the pathophysiology of inhibin A expression by the stromal cells of hemangioblastoma remains unknown and further investigation is required. PMID- 12883250 TI - Cavitary mass lesion and recurrent pneumothoraces due to Paragonimus kellicotti infection: North American paragonimiasis. AB - North American paragonimiasis is well described in omnivorous and carnivorous animals on this continent. Humans are rarely infected, largely because of dietary customs, but are at risk for infection if raw or undercooked crayfish are consumed. We describe a patient with a pleuropulmonary infection due to Paragonimus kellicotti that presented as recurrent pneumothoraces and a cavitary lesion. This is the first case of North American paragonimiasis in which the diagnosis was based on the morphology of the eggs present in histologic sections. PMID- 12883251 TI - Evidence of neural differentiation in a case of post-therapy primitive neuroectodermal tumor/Ewing sarcoma of bone. AB - Neural differentiation with the appearance of ganglion-like cells has been reported in untreated primitive neuroectodermal tumor/Ewing sarcoma (PNET/EWS) at peculiar sites, such as the cauda equina, and following treatment. The case is presented here of a 17-year-old girl with a tumor in the iliac bone. An open biopsy was diagnosed as PNET/EWS of the bone. The tumor had the typical morphology of this tumor type and showed diffuse membranous immunoreactivity for CD99, intense immunoreactivity for synaptophysin, and focal immunoreactivity for neuron-specific enolase and S-100 protein. Occasional reactivity for vimentin was evident, while no immunoreactivity for NB84a, Hu, chromogranins A and B, neurofilaments, cytokeratins, and desmin was present. The patient underwent chemotherapy and radiotherapy, followed by right internal hemipelvectomy. The post-treatment residual viable tumor showed a morphologic appearance resembling a neuroblastoma, with immunoreactivity for NB84a, Hu, synaptophysin, and chromogranins A and B, but not for CD99. RT-PCR performed on tumor tissue before and after therapy showed the presence of the EWS-FLI1 fusion transcript, type I in both samples. This case of PNET/EWS is unique in the sense of showing the typical fusion transcript associated with this tumor both in the morphologically typical pretherapy tumor and in the sample from the post-therapy specimen showing neuroblastoma-like features. PMID- 12883252 TI - Human intestinal anisakiosis due to consumption of raw salmon. AB - Anisakiosis is a parasitic infection that follows consumption of raw or insufficiently pickled, salted, smoked, or cooked wild marine fish infected with Anisakis sp. larvae. We report a case of intestinal anisakiosis in a 50-year-old man from Quebec who presented with abdominal pain and peripheral eosinophilia after eating raw wild-caught salmon from the Pacific Ocean off Canada. Abdominal CT scan showed bowel distension proximal to a segmental jejunal wall thickening, which was resected. The jejunum segment showed a localized area of serositis with mucosal edema and a submucosal abscess rich in eosinophils surrounding a parasite consistent with the third larval stage of Anisakis sp. Diagnostic morphologic characteristics included an unpaired excretory gland (renette cell), Y-shaped lateral epidermal cords, no apparent reproductive system, and a ventriculus (glandular esophagus). These features and the absence of lateral alae excluded Ascaris sp. The absence of ventricular appendage and intestinal cecum excluded other anisakids of the genera Pseudoterranova and Contracaecum. As the popularity of eating raw fish is growing in North America, anisakiosis may be diagnosed more frequently in surgical specimens. This parasitic infection should be considered in the differential diagnosis of acute abdominal syndromes and eosinophilic infiltrates of the stomach, small intestine, colon, omentum, and mesentery, especially with a history of raw marine fish consumption. PMID- 12883253 TI - Ethical issues in anatomic pathology. PMID- 12883254 TI - Papillary thyroid carcinoma. PMID- 12883255 TI - Primitive neuroectodermal tumors. PMID- 12883256 TI - Observer variation of lesions of the thyroid. PMID- 12883257 TI - Controversies in clinical pancreatology: management of acute idiopathic recurrent pancreatitis. PMID- 12883258 TI - Controversies in clinical pancreatology: should the sphincter of Oddi be measured in patients with idiopathic recurrent acute pancreatitis, and should sphincterotomy be performed if the pressure is high? AB - The risk benefit ratio of measuring the sphincter of Oddi pressure and performing sphincterotomy in patients with idiopathic acute pancreatitis with high sphincter pressures has not been assessed. Few healthy controls have had sphincter measurements so that the definition of an abnormal sphincter remains understudied. The procedures involved have significant risks and the benefits are hard to measure considering the variable natural history of this disorder. No prospective, randomized investigations have been performed to evaluate efficacy of these invasive procedures. It is the author's opinion that until appropriate studies prove efficacy that these interventions be considered experimental. PMID- 12883259 TI - Mutations in the lipoprotein lipase gene as a cause of hypertriglyceridemia and pancreatitis in Taiwan. AB - INTRODUCTION: Familial lipoprotein lipase (LPL) deficiency is inherited as an autosomal recessive trait and is characterized by chylomicronemia, eruptive xanthoma, hepatosplenomegaly, and recurrent pancreatitis. AIMS AND METHODOLOGY: Two unrelated Chinese of Han descent with hypertriglyceridemia were enrolled in this study, and another six Han Chinese with no family history of hypertriglyceridemia and diabetes were recruited as normal controls. LPL activity was determined with use of an artificial substrate of 14C-trioleine and Arabic gum, and release of 14C free fatty acid was determined by the liquid-liquid partitioning system. LPL mass was measured by enzyme immunoassay. Genomic DNA was extracted from EDTA-preserved whole blood, and PCR was used to amplify the nine coding exons and the minimal promoter of the LPL gene. RESULTS: DNA sequence analysis revealed that mutations were identified in both patients; one patient had compound heterozygous mutations in codon 252 [CTG(Leu) --> GTG(Val)] and in codon 264 [TGC(Cys) --> TGa(Ter)] of exon 6, and the other patient had homozygous L252V mutation. These subjects had > or =90% reduction in LPL mass and > or =60% reduction in LPL activity. CONCLUSION: The mutated and truncated LPLs caused hypertriglyceridemia in these patients in Taiwan with hypertriglyceridemia and pancreatitis. PMID- 12883260 TI - Dynamic aspects of granulocyte activation in rat severe acute pancreatitis. AB - We demonstrated dynamic aspects of granulocyte activation in rat severe acute pancreatitis, which was induced by cerulein and aggravated following lipopolysaccharide (LPS) injection. Pancreatitis induced by cerulein increased intracellular elastase activity of granulocytes in the blood. However, significant systemic cytokinemia was not provoked under such conditions. After induction of severe pancreatitis by LPS, intracellular elastase activity of circulating granulocytes decreased markedly and immediately. This decrease occurred simultaneous to induction of systemic hypercytokinemia and granulocyte migration into the lung. Overall results imply that: (1) circulating granulocytes are activated by induction of mild pancreatitis; (2) activation of granulocytes is mediated by factors other than systemic cytokinemia, such as locally produced cytokines; (3) those priming granulocytes immediately and significantly migrate from the circulation into the extravascular space by induction of endotoxemia; and (4) migration of granulocytes, in turn, may be mediated by systemic cytokinemia. PMID- 12883261 TI - Pancreatic fungal infection. AB - INTRODUCTION: There has been a steady rise in the incidence of pancreatic fungal infections. Even though fungal infections of the pancreas are not very common, they are often nosocomial. AIMS: To review pancreatic fungal infections and to compare their clinical characteristics with those of bacterial pancreatic infections, along with the causes and approaches to diagnosis and treatment. RESULTS: Because of the lack of randomized, prospective trials, standardized recommendations for use of antifungal prophylaxis would be premature. The most important aspect of prophylaxis against pancreatic fungal infection is minimizing the factors that predispose the pancreas to fungal infections. CONCLUSION: Isolation of fungal elements from necrotic pancreatic tissue and treatment of local infection are vital. Treatment of local candidal infection should be initiated with surgical necrosectomy. Systemic antifungal therapy should be started early in the course of the disease, but whether antifungal agents should be added to the prophylactic antibiotic regimens for patients with necrotizing pancreatitis remains questionable. PMID- 12883262 TI - Pancreatic cancer mortality in Europe: the leveling of an epidemic. AB - Mortality rates from pancreatic cancer have increased throughout Europe between the late 1950s and the 1980s. Trends in 22 European countries, the European Union (EU) and 6 selected eastern European countries have been updated using official death certification data for pancreatic cancer abstracted from the WHO database over the period 1980 to 1999. In EU men, a rise from 7.2 to 7.5/100,000 was observed between the early and the late 1980s, followed by a leveling off in the 1990s. For women, rates tended to rise up to the early 1990s, and to level off thereafter around 4.7/100,000. In eastern countries, rates for both sexes rose between the early 1980s and the mid-1990s, and leveled off thereafter around 8.5/100,000 men and 5/100,000 women. Thus, rates for men only were higher in Eastern Europe than in the EU. This analysis first documents a leveling of pancreatic cancer mortality in Europe, after decades of steady rises. This is partly or largely attributable to the decline in smoking, at least in men, but other factors, including mainly nutrition and diet, may also have played some role on these trends. PMID- 12883263 TI - Exocrine pancreatic cancer, cigarette smoking, and diabetes mellitus: a case control study in northern Italy. AB - The role of cigarette smoking and diabetes mellitus as risk factors for exocrine pancreatic cancer (PC) was investigated in a hospital based case-control study. Current smokers were at increased risk for PC (OR = 2.36, 95% CI 1.53-3.63): the magnitude of the risk was related to the lifetime amount of smoking (chi2(trend) = 17.00; P < 0.0001). Among former smokers, after 15 years from ceasing smoking, the risk for PC dropped to the level of a lifetime non-smoker, whichever the lifetime smoking amount. Diabetes was associated with a 2.89-fold increased risk for PC (95% CI 1.71-4.86): the risk was 4.76 (95% CI 1.99-11.53) for diabetes diagnosed up to 2 years before the diagnosis of PC and dropped to 2.07 (95% CI 1.02-4.20) for diabetes diagnosed more than 5 years before PC. The risk for PC was estimated according to the treatment used to control diabetes: it was 6.49 (95% CI 2.28-18.48) for insulin treated diabetes and 2.12 (95% CI 1.16-3.87) for diabetes treated with oral hypoglycemic drugs. The risk of PC for diabetes treated for more than 5 years before the diagnosis of PC was 6.21 (95% CI 1.61 23.96) for patients treated with insulin and 1.21 (95% CI 0.50-2.92) for those treated with oral hypoglycemic drugs: the type of treatment needed to control the disease may discriminate between the diabetes that represents a consequence of cancer from the diabetes that could represent an etiological co-factor. More studies are needed to clarify whether long-lasting insulin-treated diabetes is an etiological co-factor in PC. PMID- 12883264 TI - Pancreatic stellate cell activation by ethanol and acetaldehyde: is it mediated by the mitogen-activated protein kinase signaling pathway? AB - BACKGROUND: Pancreatic fibrosis is a characteristic feature of alcoholic chronic pancreatitis. Recent studies suggest that activated pancreatic stellate cells (PSCs) are the major cell-type involved in pancreatic fibrogenesis. Cultured PSCs become activated when exposed to ethanol or its metabolite acetaldehyde (as indicated by increased alpha-smooth muscle actin [alpha-SMA] expression and increased collagen synthesis). However the intracellular signaling mechanisms responsible for ethanol- or acetaldehyde-induced PSC activation remain to be fully elucidated. One of the major signaling pathways known to regulate protein synthesis in mammalian cells is the mitogen-activated protein kinase (MARK) pathway. AIMS: To examine the effects of ethanol and acetaldehyde on the MAPK pathway (by assessing the activities of the 3 major subfamilies (extracellular regulated kinases 1 and 2 [ERK 1/2], JNK and p38 kinase) in PSCs and to examine the role of p38 kinase in mediating the ethanol- and acetaldehyde-induced increase in alpha-SMA expression in activated rat PSCs. METHODS: Rat PSCs were incubated with ethanol (50 mM) or acetaldehyde (200 microM) for 15 min, 30 min, 60 min, and 24 h; and activities of ERK 1/2, JNK, and p38 kinase were assessed in cell lysates using kinase assays and Western blot. In addition, rat PSCs were treated with the specific p38 MAPK inhibitor SB203580 in the presence or absence of ethanol or acetaldehyde for 24h, and activation of the downstream protein kinase MAPKAP kinase-2 (an indicator of p38 MAPK activity) was assessed by Western blot. Specific inhibitors were also used to inhibit the activity of ERK 1/2 and JNK. Following inhibition of the above signaling pathways, alpha-SMA expression by PSCs was assessed by Western blot. RESULTS: Ethanol and acetaldehyde increased the activation of all 3 subfamilies (ERK 1/2, JNK and p38 kinase) of the MAPK pathway in PSCs. Treatment of PSCs with SB203580 abolished the ethanol- and acetaldehyde-induced increase in p38 MAPK activity and also prevented the induction of alpha-SMA expression in PSCs. However, inhibition of ERK 1/2 and JNK had no effect on ethanoland acetaldehyde-induced alpha-SMA expression in PSCs. CONCLUSIONS: (1) The MAP kinase pathway is induced in PSCs after exposure to ethanol or acetaldehyde and this induction is sustained for at least 24h. (2) The p38 MAPK pathway mediates the activation (as indicated by increased alpha-SMA expression) of PSCs by ethanol or acetaldehyde. PMID- 12883265 TI - Effects of ghrelin and other neuropeptides (CART, MCH, orexin A and B, and GLP-1) on the release of insulin from isolated rat islets. AB - BACKGROUND AND AIMS: Ghrelin, a neuropeptide containing 28 amino acids, shows a reciprocal diurnal plasma fluctuation to that of plasma insulin. The aim of this study is to clarify the dose and glucose-dependency of ghrelin on the insulin secretion and to compare its effect with that of other neuropeptides-GLP-1, CART (55-102), CART (55-76), CART (62-76), MCH, orexin A, and B. MATERIALS AND METHODS: Rat islets were incubated with 1 pmol/l-1 micromol/l of ghrelin, CART fragments, MCH, orexin A or B, or GLP-1 (n = 16-32) in the presence of 16.7 mmol/l glucose. Ghrelin (10 nmol/l) was added to islets at glucose concentrations of 3.3, 6.6, 16.7 and 25 mmol/l, respectively (n = 28-32). Also, INS-1E cells were incubated with ghrelin (1 nmol/l) in the presence of glucose (3.3, 6.6, 16.7, and 25 mmol/l). In addition, we measured the mRNA expression of the ghrelin receptor using RT-PCR. RESULTS: Ghrelin inhibited insulin secretion from islets and INS-1E cells in a dose- and glucose-dependent manner. Neither 10 pmol/l-1 micromol/l of CART fragments, MCH, orexin A, nor orexin B changed the insulin secretion at 16.7 mmol/l glucose, while GLP-1, as expected, stimulated the insulin release from rat islets. Interestingly, ghrelin receptors were expressed both in islets, INS-1E, MIN 6 and alpha cell Tca-9 lines. CONCLUSIONS: Ghrelin inhibits the insulin secretion in vitro in a dose- and glucose-dependent manner. Beta cells contain ghrelin receptors. CART fragments did not affect the insulin secretion. Ghrelin may play a physiological role for the regulation of insulin secretion. PMID- 12883266 TI - An evaluation of endogenous pancreatic enzyme levels after human islet isolation. AB - INTRODUCTION: Recent evidence has suggested that inconsistencies in human islet yield and viability after collagenase digestion is attributed to the activation of endogenous enzymes of the cadaveric donor pancreas. A study of the enzyme kinetics of serine proteases throughout human islet isolations showed a significant increase in activity levels throughout the digestion period. Following the digestion, it is important to further inhibit these enzymes by the addition of an inhibitor to the dilution media. AIM: To report the levels of endogenous pancreatic enzymes remaining after human islet isolation and the effects of three potential enzyme inhibitors on the proteases. METHODOLOGY: Human albumin, fetal calf serum, and the protease inhibitor aprotinin were incubated with the trypsin, chymotrypsin, elastase, and collagenase and were assayed for activity. RESULTS: Results at the final stage indicated that chymotrypsin retained 21.0 +/- 7.5% (mean +/- SE; n = 20) of the activity observed at the conclusion of the enzymatic digestion phase of the isolation process, whereas trypsin, elastase, and collagenase retained 3.0 +/- 1.5%, 2.1 +/- 0.6%, and 3.9 +/- 0.9%, respectively. Fetal calf serum and aprotinin showed strong inhibitory effects against bovine pancreatic trypsin; however, they showed a weak inhibitory effect against elastase. Supplementation with aprotinin failed to inhibit human chymotrypsin and elastase. Human albumin showed minimal inhibition and was shown to serve only as a competitive inhibitor. No inhibition to collagenase was observed with human albumin, fetal calf serum, or aprotinin. CONCLUSIONS: This study clearly demonstrates that low amounts of endogenous pancreatic enzymes remain active throughout the human islet isolation process and that the added inhibitors at the end of the isolation process are not fully effective at inhibiting the enzymes. PMID- 12883267 TI - Arsenic trioxide induces apoptosis in pancreatic cancer cells via changes in cell cycle, caspase activation, and GADD expression. AB - We have previously shown that arsenic trioxide blocks proliferation and induces apoptosis in human pancreatic cancer cells at low, non-toxic concentrations. The mechanisms of the apoptosis was investigated in MiaPaCa2 and PANC-1 cells that have been previously shown to be responsive to arsenic trioxide. The results show the caspase-3, caspase-7, and caspase-9 are all activated by arsenic trioxide, together with cleavage of the downstream caspase-3 target poly ADP ribose polymerase (PARP). Expression of the anti-apoptosis proteins, Bcl-2 and Mcl-1 expression decreased time-dependently while Bax expression increased. These findings indicate that the Bcl family of proteins, the mitochondrial pathway and activation of the caspase cascade are responsible for arsenic-induced apoptosis. Flow cytometric analysis revealed changes of cell cycle distribution from a G0/G1 phase arrest at 24 hours to G2/M phase arrest at 72 hours following arsenic treatment. The sub-G0/G1 cell population of apoptotic cells was increased at these times. Arsenic increased expression of the P21 protein and decreased levels of cyclin A, cyclin B1 and cyclin D1, but expression of CDK2, CDK4, CDK6, and cyclin E were not affected. Arsenic trioxide markedly enhanced the expression of GADD45 and GADD153 in a time-dependent manner. In summary, arsenic trioxide induced apoptosis in pancreatic cancer cells through activating the caspase cascade via the mitochondrial pathway, GADD expression and by modifying cell cycle progress and changes in several cycle-regulating proteins. This old drug may be valuable for treatment of pancreatic cancer. PMID- 12883268 TI - Autoantibody-mediated inhibition of pancreatic cancer cell growth in an athymic (nude) mouse model. AB - INTRODUCTION: Antiribosomal P autoantibodies are detectable in 12-16% of patients with systemic lupus erythematosus. AIM: To assess whether antiribosomal P autoantibodies could be useful as a novel form of immunotherapy for pancreatic cancer. METHODOLOGY: Three pancreatic cancer cell lines were incubated with antiribosomal P or normal human immunoglobulin. Viability was assayed with MTT, and apoptosis was detected by TUNEL. MIA PaCa-2 cells were injected into athymic mice. Animals were treated with intraperitoneal antibody or control immunoglobulin. Serum antibody levels were measured by ELISA. Tumor nodule size was measured weekly. Binding of the antibody to tumors was demonstrated with fluorescent microscopy. RESULTS: Antiribosomal P antibody inhibited pancreatic cancer cell proliferation up to 54.6% (p < 0.01) and was associated with a threefold increase in the rate of apoptosis (p < 0.05). Tumor volume after 4 weeks of treatment was 23.2 mm3, versus 141.5 mm3 for the control group (p < 0.05). Apoptosis rate within the nodules was increased twofold, to 11.4%, in comparison with control (p < 0.05). CONCLUSION: Antiribosomal P autoantibody at levels similar to those that can exist in SLE inhibits the growth of pancreatic cancer cells, in vitro and in vivo. The mechanism involves surface binding and apoptosis. PMID- 12883269 TI - Alteration of integrin expression by glial cell line-derived neurotrophic factor (GDNF) in human pancreatic cancer cells. AB - INTRODUCTION: Pancreatic cancer cells express a number of functionally active integrins that are related to their adhesive and invasive abilities. AIMS: To determine whether glial cell line-derived neurotrophic factor (GDNF) influences the expression of integrins in pancreatic cancer cell lines and to elucidate the mechanisms of adhesion and invasion to extracellular matrix (ECM) proteins. METHODOLOGY: The expression of integrin subunits and the alteration of their expression by GDNF were examined by flow-cytometric analysis and cellular enzyme linked immunosorbent assay in pancreatic cancer cell lines (MIA PaCa-2 and BxPC 3). Assays of adhesion and invasion of cancer cells to ECM proteins were conducted to investigate whether increased integrin expression affects the interaction between cancer cells and putative integrin ECM ligands. RESULTS: Expression of some of the integrin subunits in pancreatic cancer cells was enhanced by GDNF. The enhancement and associated increase in adhesive and invasive ability by GDNF were inhibited by blocking the GDNF receptor or the integrin beta1 subunit. CONCLUSIONS: In pancreatic cancer, the enhancement of integrin expression by GDNF signaling through the GDNF receptor strongly influences adhesion and invasion to ECM proteins. PMID- 12883270 TI - Hemolymphangioma of the pancreas: case report and review of the literature. PMID- 12883271 TI - A case of acute pancreatitis associated with cationic trypsinogen N29T mutation. PMID- 12883272 TI - Metastatic pancreatic malignant melanoma: tumor thrombus formed in portal venous system 15 years after initial surgery. PMID- 12883273 TI - Difference in generating mouse pancreatic epithelial cell colonies in vitro. PMID- 12883274 TI - A rapid survey of the literature indicates that most of the studies performed on isolated islets from rat, mice, or human pancreas dealt with culture conditions and secretion of the different hormones. PMID- 12883275 TI - New insulin sensitivity adjusted beta cell indices: validation according to a genetic study. AB - PURPOSE: The role of hepatocyte nuclear factor-1alpha (HNF1alpha) in the maturity onset diabetes of the young 3 (MODY3) is well established. A common polymorphism, I27L that is not linked to MODY3, has been demonstrated to affect beta cell function. To facilitate the identification of subjects with a low beta cell reserve, we developed beta cell indices and validated according to a genetic study. METHODS: Insulin sensitivity index (ISI), 1st phase insulin response (1stIR), and 2nd phase insulin response (2ndIR) were assessed in 60 glucose tolerant subjects using hyperglycemic clamps. Delta1stIR and delta2ndIR were defined as differences between 1stIR (or 2ndIR) and the ISI-adjusted 1stIR (or 2ndIR). The genotypes were determined from genomic DNA. RESULTS: Delta1stIR (P = 0.0130) and delta2ndIR (P = 0.0482) differed among the 3 genotypic groups. Multivariate analysis confirmed the independent influence of the I27L polymorphism on delta1stIR (P = 0.0130) and delta2ndIR (P = 0.0369). Within the LL group, 75% and 63% of the subjects were within the lowest quartile of delta1stIR (P = 0.0011) and delta2ndIR (P = 0.0277), respectively. CONCLUSIONS: With new beta cell indices, we demonstrated the independent impact of the I27L polymorphism on beta cell function. The new beta cell indices will facilitate the identification of glucose tolerant subjects with reduced beta cell reserve. PMID- 12883276 TI - Serum levels of mast cell tryptase, vascular endothelial growth factor and basic fibroblast growth factor in patients with acute pancreatitis. AB - PURPOSE: Mast cell tryptase, vascular endothelial growth factor (VEGF), and basic fibroblast growth factor (bFGF) possibly play a role in the pathogenesis of acute pancreatitis (AP). The aim is to describe their serum levels in relation to severity of AP. METHODS: Seventy patients with AP were studied. Thirty-one had mild acute pancreatitis and 39 severe AP of whom 21 developed organ dysfunction. Serum concentration of tryptase was determined with fluoroimmunoassay (UniCAP), and VEGF and bFGF with ELISA at admission and on days 1, 2, and 7 post hospitalization. RESULTS: The peak tryptase levels and tryptase levels at 2nd day after symptom onset, although mostly within normal range, were significantly higher in patients with organ dysfunction than in patients without organ dysfunction (6.6 microg/l (inter quartile range 4.8 to 12.6) versus 4.0 microg/l (2.7 to 6.2); P = 0.018 and 6.0 microg/l (4.4 to 7.6) versus 3.4 microg/l (2.3 to 4.8); P = 0.006, respectively). Median serum VEGF and bFGF concentrations increased during follow-up, were significantly higher on day 7 than on days 0, 1, and 2, but were not related to development of organ dysfunction. CONCLUSIONS: Mast cell activation, as defined by serum tryptase levels, may play a role in the development of remote organ dysfunction in patients with AP. However, neither tryptase nor the factors VEGF and bFGF serve as predictors of organ dysfunction in clinical AP. PMID- 12883277 TI - In vivo functioning and transplantable mature pancreatic islet-like cell clusters differentiated from embryonic stem cell. AB - INTRODUCTION: Although the differentiation of embryonic stem (ES) cells to islet like clusters using differentiation method without employing gene transfer technique has been recently reported, neither endocrine granules in the cytoplasm nor in vivo function of differentiated islet like clusters has been demonstrated. AIMS: To investigate whether ES cells could be differentiated to mature islet like clusters which show in vivo function after transplantation as well as retain endocrine granules in the cytoplasm by electron microscopic observation. METHODOLOGY: In this experiment, using mouse embryonic stem (mES) cells as a model system for lineage specific differentiation, we tried to differentiate mES cells to pancreatic islet-like cell clusters (PICCs) through a series of treatments (4-step procedure). Differentiated PICCs were analyzed and characterized by various techniques, such as RT-PCR, immunohistochemistry, electron microscopic observation, in vitro static incubation test, and in vivo transplantation to diabetic animals. RESULTS: Differentiated islet-like cell clusters from ES cells using our newly developed method (four-step procedure) showed strong expression of essential specific genes to the endocrine pancreas and also specific genes to the exocrine pancreas demonstrating that these islet like clusters were mature from the developmental biologic point of view. These differentiated cells clearly revealed many mature insulin secretory granules of pleomorphic shape in the cytoplasm as well as well-developed rough endoplasmic reticulum. In vitro study indicated that differentiated cells retain a potent insulin secretory responsiveness to glucose stimulation. Furthermore, the islet like cell clusters significantly decreased high blood glucose levels almost to normal levels when grafted to streptozotocin-induced diabetic mice without induction of any teratoma formation after transplantation. CONCLUSION: Our results provide evidence that ES cells could differentiate to functioning and transplantable mature pancreatic islet-like cell clusters using our newly developed differentiation method without employing gene transfer technique. This study may lead to a basis for production of indefinite sources of islets that could be applicable for future clinical trial. PMID- 12883278 TI - Specialized nutrition support in critically ill patients. AB - How nutritional therapy may affect real clinical outcomes is not readily apparent from a superficial reading of current data. Despite great claims in reducing infectious incidence, many studies show little difference in meaningful clinical outcomes. This has led to confusion over the role of nutrition and nutrition practice in intensive care. However, the role that nutrition plays in preventing infection and later how nutrition aids long-term recovery perhaps explain many of the misconceptions and difficulty in understanding the evidence. Encouraging new evidence is starting to show that outcome can be improved by implementing relatively simple therapies well that have an impact on nutrition and metabolic control. PMID- 12883279 TI - Treatment of metabolic acidosis. AB - Metabolic acidosis is characterized by a decrease of the blood pH associated with a decrease in the bicarbonate concentration. This may be secondary to a decrease in the strong ion difference or to an increase in the weak acids concentration, mainly the inorganic phosphorus. From a conceptual point of view, two types of nontoxic metabolic acidosis must be differentiated: the mineral metabolic acidosis that reveals the presence of an excess of nonmetabolizable anions, and the organic metabolic acidosis that reveals an excess of metabolizable anions. Significance and consequences of these two types of acidosis are radically different. Mineral acidosis is not caused by a failure in the energy metabolic pathways, and its treatment is mainly symptomatic by correcting the blood pH (alkali therapy) or accelerating the elimination of excessive mineral anions (renal replacement therapy). On the other hand, organic acidosis gives evidence that a severe underlying metabolic distress is in process. No reliable argument exists to prove that this acidosis is harmful under these conditions in humans. Experimental data even show that hypoxic cells are able to survive only if the medium is kept acidic. The management of an acute organic metabolic acidosis is therefore primarily based on the cause of the acidosis, and no scientific argument exists to justify the correction of the acid-base imbalance in this context. PMID- 12883280 TI - Antioxidant therapy in intensive care. AB - PURPOSE OF REVIEW: This review intends to summarize the recent findings regarding the presence of increased oxidative stress in critically ill patients and its potential pathophysiologic role, as well as the results of recent clinical trials of antioxidant therapies. RECENT FINDINGS: Several lines of evidence confirm the increase in oxidative stress during critical illness. The oxidative damage to cells and tissues eventually contributes to organ failure. Prophylactic administration of antioxidant vitamins or glutamine, incorporated in the nutritional support or given as separate medications, efficiently attenuates the oxidative stress and in some studies improves the outcome of critically ill patients. Few data on the effects of N-acetylcysteine or trace elements have been published during the last two years. SUMMARY: Patients at risk of organ failure could benefit from the early adjunction of antioxidant treatment, including vitamins and glutamine. PMID- 12883281 TI - Metabolic alterations in sepsis and vasoactive drug-related metabolic effects. AB - The main clinical characteristics of sepsis and septic shock are derangements of cardiocirculatory and respiratory function. Additionally, profound alterations in metabolic pathways occur leading to hypermetabolism, enhanced energy expenditure, and insulin resistance. The clinical hallmarks are hyperglycemia, hyperlactatemia, and enhanced protein catabolism. These metabolic alterations are even more pronounced during sepsis as a result of cytokine release and subsequent induction of inflammatory pathways. Increased oxygen demands from mitochondrial oxygen utilization and oxygen consumption related to oxygen radical formation may contribute to hypermetabolism. In addition, mitochondrial dysfunction with impaired cellular respiration may be present. Mainstay therapeutic interventions for hemodynamic stabilization are adequate volume resuscitation and vasoactive agents, which, however, have additional impact on metabolic activity. Therefore, beyond hemodynamic effects, specific drug-related metabolic alterations need to be considered for optimal treatment during sepsis. This review gives an overview of the typical metabolic alterations during sepsis and septic shock and highlights the impact of vasoactive therapy on metabolism. PMID- 12883282 TI - Glutamine and acute illness. AB - This is a literature review over a time period of the past 2 years concerning glutamine in clinical nutrition. Emphasis is put upon studies of glutamine in clinical settings, but a brief overview of the large range of literature over the role of glutamine in various experimental settings is also included. The most interesting concept for the past 2 years is the suggestion to use plasma glutamine concentration at admission to the intensive care unit as a prognostic marker and as a possible indicator for indication of glutamine supplementation. PMID- 12883283 TI - Antimicrobial strategies in surgical critical care. AB - There is clear evidence that early and appropriate empiric antimicrobial therapy for suspected nosocomial infections reduces the rate of adverse outcomes. This approach necessitates a liberal antimicrobial policy, whereas observational and experimental data also suggest that excessive antibiotic use promotes the emergence of antimicrobial resistance, creating a dilemma for the intensivists and begging the question as to whether minimization of antimicrobial resistance and maximization of individual patient outcomes are mutually exclusive. Contemporary strategies are outlined for the antimicrobial management of ventilator-associated pneumonia, the most common nosocomial infection in the intensive care unit, and the use of institution-specific guidelines, invasive diagnostic approaches, and other objective criteria to ensure adequate, yet not excessive use of antimicrobials are discussed. The focus is then on relative merits of routine antifungal prophylaxis as an example of an attempt to reduce the incidence and adverse consequences of late diagnoses of fungal sepsis. Finally, the advantages and disadvantages of antimicrobial cycling as a means of reducing antimicrobial resistance in the intensive care unit are outlined. PMID- 12883284 TI - Goal-oriented shock resuscitation for major torso trauma: what are we learning? AB - Shock resuscitation is an obligatory intervention for severely injured patients who present in shock. During the past 15 years, with widespread acceptance of "damage control" surgery and early triage to the intensive care unit (ICU) to optimize resuscitation, the lives of many major trauma victims have been saved, and much has been learned about shock resuscitation. Due largely to the work of Shoemaker et al., a resuscitation strategy based on a standardized process using O(2) delivery index (DO(2)I) as an endpoint and physiologic performance goal for interventions has been developed, studied, and refined for resuscitation of shock caused by major trauma. DO(2)I >or=600 mL O(2)/min-m2 is the only resuscitation endpoint variable that has been tested in prospective randomized trials (PRTs) of trauma patient outcome. These PRTs are limited, and their results are not conclusive. Results from other investigators, including our group, using similar process and endpoints, are indicating similar performance and outcomes. We believe that DO(2)I is a useful endpoint because it integrates three important variables, ie, hemoglobin concentration [Hb], arterial hemoglobin O(2) saturation, and cardiac output. We have found DO(2)I >or=500 mL O(2)/min-m2 to be an endpoint with more general applicability, but we believe that the standardized process is more important than the specific endpoint. To standardize our process, we have developed a computerized decision support tool for shock resuscitation. This technology has provided novel data collection and has permitted refinement of the bedside process. Our data analysis indicates that the next challenge will be to develop a similar pre ICU resuscitation process that will use less invasive monitors and different endpoints. Identification of the high-risk resuscitation nonresponders early in the resuscitation process will be needed to redirect their clinical trajectories. As an endpoint for interventions for goal-directed resuscitation in the critically injured trauma patient, systemic O(2) delivery is the current state of the art and the basis for near future development of clinical processes for resuscitation of shock due to major trauma. PMID- 12883285 TI - Controversy of immunonutrition for surgical critical-illness patients. AB - Specific nutrients such as arginine, glutamine, dietary nucleotides, and omega-3 fatty acids have been shown to influence infectious morbidity, antibiotic use, and hospital length of stay. The combination of these nutrients into one enteral formulation has become known as immune-enhancing diets. Consensus guidelines developed by a number of clinical investigators and published in 2001 concluded that immune-enhancing diets were beneficial in moderate to severely malnourished patients undergoing elective gastrointestinal surgery and patients sustaining severe blunt and penetrating torso trauma. The purpose of this article is to review data published between February 1, 2001, and January 31, 2003, to determine whether new studies substantiated or refuted the current recommendations for the use of immune-enhancing diets in surgical, critically injured, and critically ill patients. Results confirmed the benefits of preoperative administration of immune-enhancing diets in surgical patients but also demonstrated that postoperative administration offered no advantages. Decreased infectious complications were also observed in critically ill patients receiving immune-enhancing diets. Accumulating evidence supports the use of immune-enhancing diets in these specific patient populations. PMID- 12883286 TI - Towards better care: an exploration of some barriers and solutions to research transfer in the intensive care unit. PMID- 12883287 TI - Is the implementation of research findings in the critically ill hampered by the lack of universal definitions of illness? AB - In the design of clinical trials, a clear definition of disease is essential for enrollment of a homogeneous study population with a higher likelihood of demonstrating a benefit of an intervention. A definition that is applicable to standard clinical practice enhances the ability of clinicians to apply results of the clinical trial to patient care. Use of a universally accepted definition allows valid comparisons across multiple studies. Sepsis, the acute respiratory distress syndrome, and ventilator-associated pneumonia are examples of conditions for which universal definitions developed by panels of experts have facilitated the design of successful clinical trials. However, implementation of the results of some of these studies has been complicated by a lack of understanding or acceptance of disease definitions or by their overly inclusive nature. For example, the presence of Systemic Inflammatory Response Syndrome (SIRS) will identify most patients with sepsis, however, a significant number of patients with those clinical findings will have other underlying processes. Approved definitions for VAP are cumbersome, and adherence to those definitions in the design of clinical trials is poor. This has led to confusion regarding the accuracy of diagnostic tests and poor acceptance of evidence based guidelines by clinicians. When investigators and clinicians do not adhere to common definitions of disease, results of clinical trials may be applied inappropriately or ignored altogether. More specific identifiers of critical illnesses using specific biochemical or genetic markers are being explored. This approach may also be useful for staging disease. PMID- 12883288 TI - What does it mean to be critically ill and elderly? AB - The number of critically ill elderly continues to rise, causing health care workers to be faced with decisions regarding aggressiveness of care, rationing of resources, and optimizing outcome. Although survival rates in the critically ill elderly may be lower than those in the younger critically ill, health care workers must focus on customizing treatment to optimize physiologic recovery, quality of life, and functional status. We advocate better research designs incorporating long-term outcomes and genetic predisposition as a means of improving care in the elderly critically ill. PMID- 12883289 TI - Barriers to translating evidence into practice. AB - PURPOSE OF REVIEW: The demand for high-quality care is increasing. A variety of therapies have been shown to improve patient outcomes in critical care. Nevertheless, relatively little research has focused on identifying how to deliver those therapies effectively and efficiently. As a result, the most cost effective opportunity to improve patient outcomes will likely come not from discovering new therapies but from discovering how to deliver therapies that are known to be effective. RECENT FINDINGS: The authors review the evidence regarding changing behavior of physicians and discuss barriers to translating evidence into practice. They also provide practical examples of how they applied the concepts of teamwork, independent redundancy, and reduced complexity to change the behavior of physicians and improve quality of care. SUMMARY: Because quality is a multidimensional construct, it is unlikely that a single approach will be effective. ICU physicians and hospital leaders must assume a leadership role, implementing a combination of different approaches and developing appropriate systems for patient care. PMID- 12883290 TI - Health-related quality of life after liver transplantation: it's not all about the liver. PMID- 12883291 TI - Usefulness of an experimental model of the infant gut. PMID- 12883292 TI - Intravenous lipid emulsions in pediatrics: time for a change? PMID- 12883293 TI - Why mapping celiac disease susceptibility genes is so challenging: lessons from the CTLA-4 gene. PMID- 12883294 TI - The liver in celiac disease. PMID- 12883295 TI - Intestinal protein loss and hypoalbuminemia in children with pneumonia. AB - BACKGROUND: Intestinal protein loss has been reported mainly in diseases affecting the gastrointestinal tract. Intestinal protein loss during pneumonia with effusion has not been reported to date. The authors attempted to assess the associations between pneumonia with effusion and intestinal protein loss and hypoalbuminemia in children. METHODS: This was a prospective consecutive case series study of in children hospitalized with pneumonia and effusion during a period of 4(1/2) years. Serum albumin, C-reactive protein (CRP), and fecal alpha 1 antitrypsin (alpha-1-AT) were measured in the first 72 hours of hospitalization. Two control groups were studied: one consisted of 50 febrile children hospitalized because of viral or mild bacterial infections, and the other consisted of 20 afebrile children hospitalized because of convulsive disorders. RESULTS: Sixty-seven children ages 4 months to 14 years hospitalized with pneumonia and effusion were enrolled in the study. Fifty-nine percent (40 children) were found to have elevated fecal alpha-1-AT excretion (range, 2-10 mg/g) compared with none in the two control groups (P < 0.000).Fifty-two percent (35 children) of the children with pneumonia and effusion had mild to moderate hypoalbuminemia (range, 22-34 g/L). Only one child (2%) in the febrile control group had a low albumin of 34 g/L; none were found in the afebrile control group. The level of fecal alpha-1-AT was inversely correlated with serum albumin level. CONCLUSIONS: Pneumonia with effusion in children is often associated with an intestinal protein loss that can be monitored by measuring gastrointestinal loss of protein, namely fecal alpha-1-AT. In most cases the development of hypoalbuminemia correlates with the development of intestinal protein loss. PMID- 12883296 TI - Dietary nucleotides modulate mitochondrial function of intestinal mucosa in weanling rats with chronic diarrhea. AB - BACKGROUND: Chronic diarrhea during early infancy is characterized by intestinal mucosal injury, and as a consequence, the mitochondrial system of oxidation and reduction and energy production is altered. Since dietary nucleotides have been associated with the process of intestinal mucosal repair in rats with chronic diarrhea, the aim of this study was to examine the effects of dietary nucleotides on the functioning of mucosal mitochondria. METHODS: Weanling rats were fed with a semipurified synthetic diet (C) or the same diet in which carbohydrates were substituted by lactose (L), resulting in chronic diarrhea. During recovery, rats were fed with the semipurified synthetic diet (LC) or the same diet supplemented with nucleotides (LN). The activities of adenosine triphosphate synthase (ATPase), cytochrome c oxidase, citrate synthase, and malate dehydrogenase were measured in mitochondria from ileum and colon mucosa. RESULTS: These enzymatic activities rose in rats with chronic diarrhea, possibly to compensate for the drastic decline in adenosine triphosphate (ATP) synthesis. Dietary nucleotide supplementation allowed normalizing of the activities of ATPase (C: 0.37 +/- 0.16 microg/min/mg protein; L: 0.68 +/- 0.25 microg/min/mg protein; LC: 0.60 +/- 0.20 microg/min/mg protein; LN: 0.42 +/- 0.22 microg/min/mg protein), citrate synthase (C: 0.12 +/- 0.05 mM/min/mg protein; L: 0.21 +/- 0.07 mM/min/mg protein; LC: 0.21 +/- 0.06 mM/min/mg protein; LN: 0.12 +/- 0.02 mM/min/mg protein), and malate dehydrogenase (C: 0.77 +/- 0.48 mM/min/mg protein; L: 3.08 +/- 0.85 mM/min/mg protein; LC: 2.11 +/- 0.44 mM/min/mg protein; LN: 1.13 +/- 0.51 mM/min/mg protein) in the ileum mitochondria of the diarrheic rats. In colonic mucosa, mitochondrial enzymatic activities were restored after eliminating lactose from the diet. CONCLUSION: These results suggest that dietary nucleotides promote earlier restoration of the ileal mitochondrial function after chronic diarrhea. PMID- 12883297 TI - Increased levels of bovine serum albumin antibodies in patients with type 1 diabetes and celiac disease-related antibodies. AB - OBJECTIVES: To detect the presence of antibodies against bovine serum albumin in a cohort of Spanish patients with type 1 insulin-dependent diabetes. METHODS: Antibodies were measured using an in-house enzyme-linked immunosorbent assay test in 80 patients with type 1 diabetes, subdivided according to the presence or absence in their serum of celiac disease-related antibodies. For comparison, 30 patients with celiac disease (nondiabetic), 13 patients with autoimmune thyroiditis, and 45 healthy volunteers were used. RESULTS: Thirty-one percent of patients with diabetes yielded a positive result, with a mean value of 26.1 +/- 21.8 arbitrary units (AU). If the group was split into those with celiac disease related antibodies and those lacking them, the percentages were 53% and 25%, respectively, with a mean value of 39.6 +/- 28.4 AU and 22.4 +/- 18.3 AU (P = 0.003), respectively. Seventy-three percent of celiac patients showed bovine serum albumin antibodies with a mean level of 38.8 +/- 27.7 AU, comparable to that of patients with diabetes with celiac antibodies, but higher than the group lacking them (P = 0.001). Although 46% of patients with autoimmune thyroiditis had positive results, the level detected (22.1 +/- 8.7 AU) was significantly lower than that recorded in patients with type 1 diabetes who had celiac disease antibodies (P = 0.04) and celiac patients (P = 0.04). Healthy volunteers showed no antibodies against bovine serum albumin. CONCLUSIONS: These data suggest that bovine serum albumin antibodies appears in patients with a compromised epithelial permeability, and they reflect a general defect in the process of immunologic tolerance associated with a predisposition to autoimmunity, rather than immunity specific to beta cells. PMID- 12883298 TI - Natural history of intestinal failure, investigated through a national network based approach. AB - OBJECTIVES: Intestinal failure (IF) is a condition whose treatment requires advanced knowledge and techniques and a multidisciplinary approach. Intestinal failure is the endpoint of many intestinal diseases and may result in full recovery, in life-long parenteral nutrition, or in the death of the child. The aim of this study was to evaluate the natural history of IF in children using a national network of resources. METHODS: Italian centers of pediatric gastroenterology merged in a national network, developing a collaborative management approach for children with IF. A consensus definition of IF was achieved. A database was set up to investigate the cause, epidemiologic factors, and natural history of IF. RESULTS: One hundred nine children were enrolled in 5 years. The cause of IF was: short bowel syndrome (n = 48), disorders of motility (n = 16), structural enterocyte defects (n = 14), multiple food intolerance (n = 10), autoimmune enteropathy (n = 7), and others or unknown (n = 14). The eventual outcome was closely related to the primary cause, ranging from full and permanent intestinal sufficiency in children with multiple food intolerance to high death rate or total dependance upon parenteral nutrition in those with structural enterocyte defects. Intermediate outcomes were observed for the other causes. Four children received intestinal transplantation. CONCLUSIONS: The network approach for IF provides an effective model to optimize resources and to investigate prospectively the natural history of IF. Based on the work on this series, a European network for IF could be an effective model for fulfilling the diagnostic and management needs, including intestinal transplantation. PMID- 12883299 TI - No association of CTLA4 gene with celiac disease in the Basque population. AB - BACKGROUND: Celiac disease (CD) is an autoimmune disorder caused by intolerance to ingested gluten that develops in genetically susceptible individuals. The contribution of human leukocyte antigen (HLA) genes to the genetic risk to CD has been known for a long time; however, non-HLA genetic factors are likely to be required for the development of the disease. Several studies have associated the CD28/CTLA4 region on chromosome 2q33 with the disease in different populations. The CTLA4 gene encodes a receptor involved in the control of T-cell proliferation and mediates T-cell apoptosis. AIM To determine the contribution of two polymorphisms of the CTLA4 to the disease: the A/G dimorphism at position +49 in exon 1 and the (AT)(n) microsatellite in the 3' untranslated region. PATIENTS: Forty-one celiac families of Basque origin (43 patients with CD and 80 first degree relatives). METHODS: Restriction enzyme digestion of polymerase chain reaction amplified genomic DNA for the A/G dimorphism and polymerase chain reaction followed by high-resolution electrophoresis for the (AT)(n) microsatellite. For disease association studies, the Affected Family Based Controls approach was used. RESULTS: The frequency of the A allele of 49 A/G polymorphism was 67.47% in the celiac allele group compared with 70.13% in the Affected Family Based Controls group. These differences were not significant. Analysis of the (AT)(n) polymorphism identified 17 different alleles, ranging from 262 to 312 bp in length, but no allele was significantly associated with the disease. CONCLUSIONS: Our results did not show any evidence of association of any of the CTLA4 gene polymorphisms with the disease. This may result from population specific differences in genetics and environmental susceptibility to CD. PMID- 12883300 TI - Propofol versus midazolam plus meperidine for sedation during ambulatory esophagogastroduodenoscopy. AB - OBJECTIVE: To evaluate the efficacy and safety of propofol and meperidine plus midazolam for sedation during esophagogastroduodenoscopy (EGD) in children. METHODS: Data were collected prospectively and retrospectively from neurologically intact children (0.2-17.7 years of age) who underwent ambulatory diagnostic EGD during a 4-year period. Data were included from 155 consecutive patients receiving propofol with or without premedication with midazolam (PM group). One hundred five consecutive patients who received sedation with a midazolam plus meperidine combination served as a comparison (MM group). Outcome variables were: time required for induction of sedation, length of procedure, time for recovery, need for additional supportive measures, and need for physical restraint. RESULTS: The onset of sedation was faster and the length of procedure and recovery were significantly shorter in the PM group as compared with the MM group (P < 0.01). Patients in the MM group required restraint more often than in the PM group. A higher dose of meperidine and midazolam was used in the prospective study. This led to deeper sedation but increased need for additional support. CONCLUSIONS: Propofol is safe and effective for facilitating EGD in children. PMID- 12883301 TI - Dermatologic manifestations of Crohn disease in children: response to infliximab. AB - Dermatologic extraintestinal manifestations of Crohn disease may be refractory to treatment with corticosteroids and immunomodulators. The authors describe four children with Crohn disease with dermatologic manifestations: pyoderma gangrenosum, orofacial involvement, erythema nodosum, and idiopathic lymphedema. These dermatologic conditions were unresponsive to conventional therapy but had rapid and sustained response to the anti-TNF-alpha antibody infliximab. No adverse reactions occurred. Infliximab should be considered for treating the extraintestinal dermatologic manifestations of Crohn disease in children. PMID- 12883302 TI - Functional outcomes of pediatric liver transplantation. AB - The functional status and health-related quality of life (HRQOL) of children who survive liver transplantation (LT) have not been well documented. The purpose of this study was to determine the functional status and HRQOL in this population using a validated measure for children, the Child Health Questionnaire-Parent Form 50 (CHQ-PF50). METHODS: The CHQ-PF50 instrument was completed by the parents of 55 children who agreed to participate in a mailing survey. Subscale scores for the sample were compared with those of a published normal population (n = 391). RESULTS: Study sample characteristics were: 87% Caucasian, 54.5% female, mean age at survey was 9.6 years (range, 5-17 years). Responding caregivers were 95% biologic parents and 93% female. Compared with the normal population, LT recipients had lower subscale scores for general health perceptions (P < 0.0005), emotional impact on parents (<0.0005) and disruption of family activities (0.0005). The mean physical summary score of the LT recipients was lower than that of the normal population 48.1 +/- 12.1 (P = 0.005), but the mean psychosocial summary score was similar 48.8 +/- 11.9 (P = 0.156). Within the LT population, the original diagnosis (biliary atresia vs. other), type of LT (living donor vs. cadaveric), age at LT, z score for height, and hospital days did not significantly influence any of the subscale scores. CONCLUSIONS: Children who have survived LT have functional outcomes in the physical domain that are lower than those of normal children. Self-esteem and mental health in this group appeared normal. The parents in this sample experienced more emotional stress and disruption of family activities than did parents in a normal population. PMID- 12883303 TI - Parenteral fat emulsions based on olive and soybean oils: a randomized clinical trial in preterm infants. AB - OBJECTIVE: To evaluate in premature infants a new parenteral lipid emulsion based on olive and soybean oils (ratio 4:1), with less polyunsaturated fatty acids (PUFA) and more alpha-tocopherol than standard soybean oil emulsion. STUDY DESIGN: Premature infants (gestational age, 28-<37 weeks) were randomized to receive one of the two emulsions within the first 72 hours of life. The triglyceride dose was increased to 2 g/kg/day within 3 days. Plasma phospholipid fatty acids, alpha-tocopherol/lipid ratio, and urinary malondialdehyde (MDA) excretion were determined at baseline and after 7 days. RESULTS: Of 45 recruited infants, 33 completed the study per protocol (15 soybean oil, 18 olive oil emulsion). At study end, groups did not differ in plasma phospholipid arachidonic acid, total n-6 and n-3 metabolites, but the olive oil group showed higher values of the PUFA intermediates C18:3n-6 (0.19% +/- 0.01% vs. 0.13% +/- 0.02%, P < 0.05) and C20:3n-6 (2.92% +/- 0.12% vs. 2.21% +/- 0.17%, P = 0.005). The plasma alpha-tocopherol/total lipd ratio was higher in the olive oil group (2.45 +/- 0.27 micromol/mmol vs. 1.90 +/- 0.08 micromol/mmol, P = 0.001), whereas urinary MDA excretion did not differ. CONCLUSION: The lower PUFA supply with the olive/soybean oil emulsion appears to enhance linoleic acid conversion. The reduced PUFA content, combined with a higher antioxidant intake in the olive oil group, results in an improved vitamin E status. The olive oil-based emulsion is a valuable alternative for parenteral feeding of preterm infants who are often exposed to oxidative stress, while their antioxidative defense is weak. PMID- 12883304 TI - A human flora-associated rat model of the breast-fed infant gut. AB - OBJECTIVES: Bacterial colonization of the infant gut may have important influences on the development of gastrointestinal, respiratory, and allergic disease. Early diet is a major determinant of the gut microflora. It is very difficult to carry out studies in human infants that can investigate the interaction of diet, flora, and mucosa. In this study we have developed an infant human flora-associated (IHFA) rat model to allow such investigation. METHODS: Germ-free infant rats were infected with fecal bacteria from exclusively breast fed infants and were maintained on a modified infant formula for 8 weeks. The fecal and cecal contents were collected and compared with feces of breast-fed infants for bacterial populations, bacterial metabolites, and enzymes and for the ability to inhibit adhesion of pathogenic bacteria to human mucosal cells. RESULTS: The IHFA cecum and feces were dominated by lactic acid bacteria, Bifidobacterium, and lactobacilli, which were representative of the infant feces. The fecal short-chain fatty acid profile was dominated by acetic and lactic acid in a similar manner to human infant feces. Other bacterial metabolites were similar to those of the human infant. Rat intestinal samples were able to inhibit the adhesion of pathogens to mucosal cells, but to a lesser extent than the human samples. CONCLUSIONS: This IHFA infant model of the intestinal flora of the breast-fed infant is considered valid for studying the effect of diet on bacterial colonization and metabolism. PMID- 12883305 TI - Stimulation with fatty meal (Lipomul) to assess gallbladder emptying in children with chronic acalculous cholecystitis. AB - OBJECTIVE: Chronic acalculous cholecystitis previously has been diagnosed by hepatobiliary scan stimulated with intravenous octapeptide of cholecystokinin. This compound may soon be unavailable in the United States. The goal of this study was to describe the characteristics of children with chronic acalculous cholecystitis diagnosed by hepatobiliary scintigraphy with oral Lipomul challenge, and to evaluate their clinical response to cholecystectomy. METHODS: Retrospective chart review of patients with no gall stones detected by abdominal ultrasound or computed tomography with gallbladder ejection fraction (GBEF) <35% after Lipomul challenge who subsequently underwent cholecystectomy. Fifteen patients with a mean age of 14.9 +/- 0.9 years were included. The mean duration of symptoms before evaluation was 8.2 +/- 2.5 months. RESULTS: The mean GBEF after Lipomul challenge was 16.7% +/- 2.7%. All patients had abdominal pain. Ninety percent had right upper quadrant pain and 86% had typical biliary colic. Pain was precipitated by fatty meals in 73.3%. Histopathologic analysis of the gallbladder demonstrated chronic cholecystitis in 80% of cases. The mean postoperative follow-up was 20 +/- 5 months. Six months after the surgery, nine patients (60%) were asymptomatic, five (33%) had marked improvement of symptoms, and one (6%) was unchanged. At the time of latest follow-up, symptoms had reappeared in two patients who had been asymptomatic at the 6-month visit (13%). Seven patients (46%) remained asymptomatic, five (33%) had marked improvement but continued to have some persistent symptoms, and one (6%) was unchanged. CONCLUSIONS: Chronic acalculous cholecystitis may be responsible for right upper quadrant pain in children without gallstones. A GBEF <35% at 30 minutes after Lipomul challenge may be useful in identifying patients who could benefit from cholecystectomy. Lipomul may be a good alternative to cholecystokinin for gallbladder stimulation during scintigraphy. PMID- 12883306 TI - Varicella in pediatric liver transplant patients: a retrospective analysis of treatment and outcome. AB - BACKGROUND: Varicella is a common childhood disease that can cause morbidity and mortality among immunosuppressed patients. There have been few previous studies monitoring the course of pediatric liver transplant patients with acute varicella. The aim of this study was to evaluate the treatment, outcomes, and complications of pediatric liver transplant patients admitted with acute varicella infection. METHODS: A retrospective chart review was carried out based on discharge diagnoses of orthotopic liver transplant and varicella among pediatric patients (age range, birth-18 years) admitted to the UCLA Medical Center between 1985 and 2001. RESULTS: Five hundred fifty-six pediatric patients received liver transplantations between 1985 and 2001. Twenty-two of these patients were admitted to the UCLA Medical Center with varicella (11 females, 11 males). No patients were treated on an outpatient basis. Mean age of the patients was 6 years (range, 1-16 years). None of these patients received the varicella vaccine before hospitalization. On admission, 5 of 22 patients (23%) had received varicella zoster immunoglobulin within 96 hours of exposure. The mean length of hospitalization was 6 days (range, 2-11 days). All immunosuppression dosages were reduced during the admissions. None of the patients had been treated with high dose corticosteroids for acute rejection before the onset of the varicella infection. Patients were treated until defervescence with intravenous acyclovir and until their varicella lesions crusted. Patients were discharged with oral acyclovir to complete a 10-day course (including the intravenous treatment). No patients had complications from the varicella infection. A complication of an elevated serum creatinine for one patient was noted with the intravenous acyclovir treatment. This patient had associated headache and nausea that resolved when the creatinine level returned to normal. CONCLUSIONS: There were no complications or dissemination of varicella infection among our pediatric liver transplant patients. Further prospective randomized trials are required to evaluate the management of pediatric liver transplant patients infected with varicella. PMID- 12883307 TI - Plasma markers of platelet activation in cystic fibrosis liver and lung disease. AB - BACKGROUND: Transforming growth factor beta 1 (TGFbeta1) is a major fibrogenic cytokine, the expression of which is increased in the livers of children with cystic fibrosis liver disease (CFLD) and in the bronchoalveolar lavage fluid of patients with cystic fibrosis pulmonary disease (CFPD). The purpose of our study was to investigate the usefulness of plasma TGFbeta1 as a noninvasive marker of CFLD and CFPD, or both and to investigate the contribution of platelet-derived TGFbeta1 to plasma TGFbeta1 by correlating the latter with platelet factor 4 (PF4). METHODS: Three groups of patients with cystic fibrosis were studied: 1) those with CFLD, 2) those with CF and no liver disease (CFNLD), and 3) those with CFPD. Controls were healthy adolescents and adults. Plasma TGFbeta1 was assayed using the R and D Quantikine quantitative sandwich enzyme immunoassay technique and PF4 (American Bioproducts ELISA kit). RESULTS: Plasma TGFbeta1 was markedly increased in CFPD (15 +/- 3 ng/mL) versus healthy adults (1 +/- 0 ng/mL; P < 0.004. The PF4 values followed a similar pattern: 105 +/- 8 ng/mL in CFPD versus 12 +/- 4 ng/mL (P < 0.0001). Plasma TGFbeta1 values in CFLD did not differ from CFNLD patients of comparable age nor from values for healthy adolescents. Plasma TGFbeta1 values were strongly correlated with values for PF4: r = 0.543, P < 0.0001. CONCLUSIONS: Plasma TGFbeta1 is not a useful marker of CFLD. The increased plasma TGFbeta1 and PF4 in CFPD patients are of interest both as possible noninvasive markers of pulmonary fibrosis and because the increased values suggest that platelet activation may play a pathogenetic role in CFPD. PMID- 12883308 TI - Therapy for pulmonary aspiration of a polyethylene glycol solution. PMID- 12883309 TI - Hemorrhagic bowel necrosis associated with acute digitalis poisoning in an infant. PMID- 12883310 TI - Loperamide suppositories in an adolescent with childhood-onset functional non retentive fecal soiling. PMID- 12883311 TI - Intrapyloric botulinum toxin injection improves gastric emptying. PMID- 12883312 TI - Results of a new technique in the treatment of severe esophageal stenosis in children: poliflex stents. AB - OBJECTIVES: The authors present their experience in treating cases of severe esophageal stenosis resistant to conventional dilatation through the placement of silicone stents, describe a precise placement technique, and establish a standard protocol for the use of these devices. METHODS: The authors detail the placement procedure and follow-up in a series of 10 patients (5 male, 5 female) between 1 and 23 years, in whom the technique was used. Nine patients had severe esophageal scarring caused by alkaline agents, and another had severe stenosis resulting from esophageal atresia; all were included in the dilatation program from the age of 6 months to 19 years. The method employs a self-expanding biocompatible stent that is placed using a special applicator over the stenotic area after performing a dilatation. This is maintained long enough to guarantee esophageal scarring without stenosis, permitting oral feeding of the patient. In general, the children experienced nausea and some vomiting during periods lasting from 12 to 48 hours; these episodes were more common when the lesion was higher and longer. Treatment with midazolam and ondansetron relieved the symptoms. All patients received omeprazole while the stent was in place. RESULTS: Five patients received one stent, four others required two, and another needed three because of restenosis. In the five patients who required stent replacement, a low dose of steroids (0.3 mg/kg/24 h) was administered 2 weeks before and also during the placement time. Currently, five patients are completely cured, with follow-up ranging from 4 to 19 months. Three patients now require stent replacement, and two others are under observation after having the stent removed. Long-term tolerance to the stent has been excellent in all cases. The time the stent was in place varied from 20 to 133 days. CONCLUSIONS: The siliconated polypropylene stents (Poliflex/Rusch) appear to be a promising new tool in the conservative treatment of esophageal stenosis. The stents permit normal feeding during application, substantially reduce treatment time, and avoid repeated anesthesia sessions for dilatations. PMID- 12883313 TI - Large-volume paracentesis and liver transplantation. PMID- 12883314 TI - Treatment of obese children and adolescents in Germany. PMID- 12883315 TI - Measuring esophageal distances. PMID- 12883316 TI - Sympathoinhibitory and depressor effects of amlodipine in spontaneously hypertensive rats. AB - The authors examined whether central actions contribute to the hypotensive effects of peripherally administered amlodipine, a lipophilic dihydropyridine with slow onset and long duration of action. After 5 to 6 weeks of high (8%, H Na) or regular (0.6%, R-Na) salt intake, changes in renal sympathetic nerve activity (RSNA), mean arterial pressure (MAP), and heart rate (HR) were recorded at rest and in response to intravenous (iv) and intracerebroventricular (icv) injection, and prolonged iv infusion of amlodipine, in conscious spontaneously hypertensive rats (SHR). Iv injection of amlodipine at 50 to 100 microg/kg decreased MAP but increased RSNA and HR in a dose-related manner. In contrast, icv injection of amlodipine at 10 to 50 microg/kg caused parallel decreases in MAP, RSNA, and HR. Iv infusion of amlodipine at 50 microg/kg per hour for 3 hours followed by 100 microg/kg per hour for 2 hours also decreased in parallel RSNA, MAP, and HR. Maximal decreases in RSNA, MAP, and HR in response to icv injection and iv infusion were significantly larger in SHR on H-Na versus R-Na. All responses lasted at least 1 hour following iv and icv injection, or after the termination of iv infusion of amlodipine. These data suggest that in SHR during prolonged iv infusion, amlodipine appears to cross the blood-brain barrier, block brain l-type Ca2+ channels, and decrease sympathetic outflow and thereby BP. Central actions may prevail during iv infusion of amlodipine at low rates, and the decrease in BP is associated with sympathoinhibition. High salt intake markedly enhances its sympathoinhibitory action, likely through central mechanisms. PMID- 12883317 TI - Verapamil, a Ca2+ entry blocker, targets the pore-forming subunit of cardiac type KATP channel (Kir6.2). AB - This study investigated the mechanism by which verapamil, which blocks 10R1, l type Ca2+ channel and the HERG channel, blocks ATP-sensitive K+ (K(ATP)) channels. In whole cell patch experiments, verapamil reversibly inhibited cardiac type K(ATP) (Kir6.2/SUR2A) channels previously activated by 100-micromol/L pinacidil. In inside-out patch experiments, verapamil inhibited the C-terminal truncated form of Kir6.2 (Kir6.2DeltaC36) in a concentration-dependent manner; half-maximal inhibition (IC(50)) was obtained at 11.5 +/- 2.8 micromol/L when Kir6.2DeltaC36 was expressed without SUR2A. Verapamil also inhibited Kir6.2/SUR2A with a similar potency; IC(50) was 8.9 +/- 2.1 micromol/L for Kir6.2/SUR2A (not statistically different from the value for Kir6.2DeltaC36 alone). Thus, verapamil appeared to target the pore-forming subunit Kir6.2 rather than SUR2A, a member of ABC superfamily. Verapamil did not decrease the single-channel conductance, but increased the closed time of Kir6.2/SUR2A. The mutations of Kir6.2DeltaC36 (Kir6.2DeltaC36-R50G, -K185Q, -G334D), which have much lower ATP sensitivity, had no significant effect on verapamil block, suggesting that the site at which verapamil mediates K(ATP) channel inhibition is not identical with that involved in ATP block. PMID- 12883318 TI - Hemodynamic effects of a new inotropic compound, PST-2744, in dogs with chronic ischemic heart failure. AB - Inotropic agents for acute decompensated heart failure are associated with a lack of efficacy or increased mortality. New compounds are needed to support patients with acute exacerbations of heart failure. This study examined the hemodynamic effects of a new inotropic agent (PST-2744) in dogs with chronic ischemic heart failure. Eight mongrel dogs at low risk for postmyocardial infarction (MI) sudden death entered the protocol. Dogs were studied after ischemic left ventricular dysfunction was induced by repeated injections of latex microspheres into the circumflex artery until the ejection fraction reached 35%. Hemodynamic parameters were measured at baseline and peak drug effect (PST-2744 5 microg.kg-1.min-1). In 5 animals, PST-2744 effects were compared with dobutamine. Heart rates, PR intervals and QT intervals were unchanged following PST-2744 administration. PST 2744 increased contractility (+dP/dt) by 56% from 1881 +/- 282 mm Hg/s to 2939 +/ 734 mm Hg/s (P < 0.01). The inotropic effect of PST-2744 was equal to that produced by 5-microg.kg-1.min-1 dobutamine (56% increase in +dP/dt), but peak heart rates were significantly higher with dobutamine (129 +/- 24 bpm PST-2744 versus 160 +/- 6 bpm 5-microg.kg-1.min-1 dobutamine, P < 0.002). No arrhythmias or conduction delays were seen with either compound. PST-2744 is an effective inotropic agent without positive chronotropic effect in subjects with stable moderate left ventricular dysfunction. PMID- 12883319 TI - Inhibition of matrix metalloproteinases prevents cardiac hypertrophy induced by beta-adrenergic stimulation in rats. AB - Insulin-like growth factor (IGF) -I is one of the candidates for cardiac hypertrophy induced by beta-adrenergic stimulation. However, the mechanisms by which the biologic actions of IGF-I are regulated under this condition remain unclear. IGF-I becomes bioavailable for its receptors upon its dissociation from IGF-binding protein (IGFBP) through IGFBP degradation. Because matrix metalloproteinases (MMPs) have been implicated in the degradation of IGFBPs, the authors investigated the role of MMPs in the regulation of the IGF-I action through the degradation of IGFBPs in cardiac hypertrophy induced by beta adrenergic stimulation. They examined the expression of MMPs in cardiac tissues of rats infused with isoproterenol (3 mg/kg per day), the effect of a MMP inhibitor, SI-27 (5 mg/rat per day), on cardiac hypertrophy, and the expression of IGF-I and IGFBP-3. MMP-1 and -2 activities increased and IGFBP-3 was degraded in heart hypertrophied by isoproterenol. MMP inhibition caused a regression in the myocyte hypertrophy in association with the suppression of both IGF-I protein in myocytes and the degradation of IGFBP-3 protein. These results suggest that the induction of myocyte hypertrophy by isoproterenol is mediated, at least in part, by a modulation of the IGF-I axis. PMID- 12883320 TI - Assessment of cardiac function by echocardiography in conscious and anesthetized mice: importance of the autonomic nervous system and disease state. AB - In this study, the authors sought to evaluate the mechanisms responsible for echocardiographically determined differences in cardiac structure and function between conscious and anesthetized mice to determine whether such differences were more or less evident in diseased states. Cardiac parameters were determined by transthoracic echocardiography. Mice anesthetized with a mixture of ketamine and xylazine showed reductions in heart rate (HR, 252 +/- 16 beats/min versus 734 +/- 9 beats/min) and fractional shortening (FS, 35% +/- 2% versus 59% +/- 2%) compared with conscious mice. Conscious mice responded little to the beta-agonist isoproterenol or atropine, but showed profound reductions in HR and FS in response to the beta(1)-antagonist atenolol. In contrast, both isoproterenol and atropine led to increases in HR and FS in anesthetized mice. The stress in conscious animals was reduced by the sedative midazolam, leading to partial restoration of responses to isoproterenol. Mice with constitutive activation of the beta-adrenergic system, due to cardiac overexpression of beta(2)-adrenergic receptors or with heart disease (myocardial infarct and pressure-overload hypertrophy) showed few differences in functional parameters between conscious and anesthetized states, attributable to pre-existing activation of the sympathetic and beta-adrenergic systems, even during anesthesia. The results indicate that the autonomic nervous system plays a critical role in the observed differences in cardiac structure and function between anesthetized and conscious mice. PMID- 12883321 TI - Elevated levels of the serum endogenous inhibitor of nitric oxide synthase and metabolic control in rats with streptozotocin-induced diabetes. AB - This study was designed to determine the relationship between elevated levels of the endogenous inhibitor of nitric oxide synthase, asymmetric dimethylarginine (ADMA), and metabolic control in rats with streptozotocin-induced diabetes. Serum levels of ADMA were measured by high-performance liquid chromatography at 8 weeks after diabetes was induced. Endothelium-dependent relaxation to acetylcholine was tested in aortic rings from nondiabetic age-matched control, untreated diabetic, and insulin-treated diabetic rats to evaluate endothelial function. Serum concentrations of glucose, glycosylated serum protein, and malondialdehyde were examined to estimate metabolic control. Serum levels of ADMA increased dramatically in untreated diabetic rats compared with control rats. This elevation in ADMA levels was accompanied by impairment of the endothelium dependent relaxation response to acetylcholine in aortic rings. Long-term insulin treatment not only prevented the elevation of serum ADMA levels, but also improved the impairment of endothelium-dependent relaxation in diabetic rats. Serum levels of glucose, glycosylated serum protein, and malondialdehyde were significantly increased in parallel with the elevation of ADMA in untreated diabetic rats compared with control rats. These parameters were normalized after diabetic rats received insulin treatment for 8 weeks. These results provide the first evidence that an elevation in the concentration of ADMA in rats with streptozotocin-induced diabetes is closely related to metabolic control of the disease. PMID- 12883322 TI - Exogenous biopterins requirement for iNOS function in vascular smooth muscle cells. AB - The authors evaluated whether supplementation of tetrahydrobiopterin (BH4) or the BH4 precursor in vascular smooth muscle cells (VSMC) that are deficient in de novo BH4 production affected the ability of these cells to synthesize nitric oxide (NO). GTP cyclohydrolase I (GTPCH) is the rate-limiting enzyme for the de novo synthetic pathway of BH4. The selective GTPCH inhibitor 2,4-diamino-6 hydroxypyrimidine (DAHP) restricts the de novo synthesis of BH4 in VSMC. Thus, treatment with DAHP and cytokines should lead to the intracellular accumulation of inactive inducible nitric oxide synthase (iNOS) apoenzyme (relative to BH4), which can be reactivated upon the repletion of BH4. Using such VSMC deficient in de novo BH4, the authors evaluated their ability to synthesize NO by supplementing with BH4 or the BH4 precursor sepiapterin or dihydrobiopterin. Adding BH4 to VSMC increased NO production in a concentration-dependent manner. Sepiapterin (SEP) or 7,8-dihydrobiopterin (BH2) also dose-dependently induced NO generation. Nitric oxide was produced in the order SEP >BH2>> BH4 at half-maximal concentrations for stimulation of 0.05, 0.1, and 1 micromol/L, respectively. Addition of BH4 or SEP substantially induced iNOS enzyme activity, which was quantified as the formation of [3H]L-citrulline from [3H]L-arginine. SEP or BH2 in the presence of methotrexate, a selective inhibitor of dihydrofolate reductase, no longer induced NO production. In contrast, although the amount of NO induced by supplemented BH4 was substantially depressed, higher concentrations of BH4, but not SEP or BH2, significantly caused NO production even in the presence of methotrexate. Thus, BH4 produced from BH2 is largely responsible for NO production in VSMC that are deficient in de novo BH4. NOS would preferentially use BH4 that is regenerated from BH2 via dihydrofolate reductase, rather than BH4 or BH2 obtained from the cytosolic milieu. PMID- 12883323 TI - Effects of simvastatin on endothelial function after chronic inhibition of nitric oxide synthase by L-NAME. AB - Blood pressure, plasma NO(2) and NO(3) level, heart weight index, antioxidant enzyme activity, and vascular reactivity in rat intact aortic rings were assessed to investigate the effects of 8-week treatment with the hydroxy-methyl-glutaryl coenzyme A reductase inhibitor simvastatin (1 mg/kg per day) on endothelial dysfunction induced by chronic Nomega-nitro-l-arginine methyl ester (l-NAME 70 mg/kg per day). Results were compared with those obtained in rats receiving l NAME, simvastatin or control animals. Coadministration of simvastatin did not restore l-NAME-increased blood pressure but normalized heart weight index (P < 0.05), endothelium-dependent relaxation to acetylcholine (P < 0.001), and plasma NO(2) and NO(3) concentration (P < 0.001) without affecting relaxation to sodium nitroprusside. Endothelium-dependent relaxation in these animals was abolished by acute incubation with l-NAME, unaffected by thromboxane synthetase inhibitor and TXA(2)/PGH(2) receptor antagonist, ridogrel, and decreased by indomethacin. Simvastatin treatment also increased plasma NO(2)+NO(3) without affecting endothelial function, heart weight index, and blood pressure of control rats. The presence of superoxide dismutase (SOD) and catalase improved endothelial relaxation only in l-NAME-treated rats, but O(2)- generated by hypoxanthine and xanthine oxidase inhibited the relaxant effect in both l-NAME and simvastatin plus l-NAME-treated rats. SOD activity was increased in all groups receiving simvastatin. Long-term treatment with simvastatin restored l-NAME-induced endothelial dysfunction, probably by preventing nitric oxide decrease. Other effects of simvastatin, including release of compensating vasodilatory cyclo oxygenase products and increased SOD activity, could also be involved. PMID- 12883324 TI - Vitamins C and E improve endothelial dysfunction in intrauterine-undernourished rats by decreasing vascular superoxide anion concentration. AB - Epidemiological studies suggest that intrauterine undernutrition plays an important role in the development of arterial hypertension in adulthood. Ascorbic acid (vitamin C) and alpha-tocopherol (vitamin E) have antioxidant properties that could improve redox-sensitive vascular changes associated with hypertension. The authors determined whether vitamins C and E treatments ameliorate the hypertension and vascular function in male rats submitted to intrauterine undernutrition. Pregnant Wistar rats were fed either normal or 50% of the normal intake diets during the whole gestational period. At 14 weeks of age, male offspring of nutritionally restricted dams were divided into 3 subgroups: vehicle treated (vehicle for 15 days, by gastric gavage, n = 9), vitamin C-treated (ascorbic acid, 150 mg/Kg/d for 15 days, by gastric gavage, n = 15) and vitamin E treated (alpha-tocopherol, 350 mg/kg per day for 15 days, by gastric gavage, n = 15). Systolic blood pressure was determined before and after antioxidant treatments by the tail-cuff method. At 16 weeks of age, the rats were used for the study of microvascular reactivity and intravital fluorescence microscopy. Intrauterine undernutrition induced hypertension, and vitamins C or E treatments reduced the blood pressure levels. The decreased acetylcholine and bradykinin induced vasodilation was restored in the vitamin-treated rats. These effects were associated with decreased vascular superoxide anion concentration. The results show that vitamins C and E reduce oxidative stress and high blood pressure levels, and improve vascular function in intrauterine-undernourished rats. PMID- 12883325 TI - Voltage-dependent effects of barnidipine in rat vascular smooth muscle. AB - The effects of the dihydropyridine nifedipine and its more lipophilic congener, barnidipine, were investigated in smooth muscle preparations from the rat in resting and depolarizing conditions. Both drugs relaxed precontracted aortic rings more potently in depolarizing conditions, barnidipine being more potent than nifedipine. Currents through Ca2+ channels in rat vascular smooth muscle cells (A7r5) and in isolated rat cardiomyocytes were reduced more potently by both drugs at a holding potential of -40 mV than at -80 mV. However, barnidipine and nifedipine were more effective in reducing the current in A7r5 cells than in cardiomyocytes. The IC(50) obtained in aortic rings and in A7r5 cells were similar for barnidipine but an order of magnitude different for nifedipine. The results show that, in depolarizing conditions, barnidipine was more effective than nifedipine. It is suggested that the higher potency of barnidipine acting in vascular smooth muscle is related to both a higher affinity to the inactivated state of vascular Ca2+ channels and to a more lipophilic property as compared with nifedipine. PMID- 12883326 TI - Effects of antisense peptide nucleic acid to platelet-derived growth factor A chain on growth of vascular smooth muscle cells. AB - To investigate antisense peptide nucleic acid (PNA) as a gene therapy for the arterial proliferative diseases, the authors designed and examined the effects of an antisense PNA targeting platelet-derived growth factor (PDGF) A-chain on expression of PDGF A-chain and growth of vascular smooth muscle cells (VSMCs) from spontaneously hypertensive rats. A 15-mer antisense PNA complementary to the initiation codon of rat and human PDGF A-chain mRNA was synthesized and purified by high-performance liquid chromatography. Gel-shift assay and biomolecular interaction analysis (BIAcore) revealed that the antisense PNA bound weakly to the target RNA, whereas it bound strongly to the target DNA. Fluorescein isothiocyanate-labeled antisense PNA to PDGF A-chain was taken up slowly and maintained in VSMCs for a prolonged period of time. Antisense PNA inhibited expression of PDGF A-chain mRNA and protein as well as DNA synthesis in VSMCs in a dose-independent manner. Inhibition of DNA synthesis by the antisense PNA was greater than that by the antisense DNA at a low concentration (0.5 micromol/L). These results suggest that antisense PNA to PDGF A-chain will be used as a gene therapy for vascular proliferative diseases such as hypertensive vascular diseases, restenosis of coronary arteries after angioplasty, and atherosclerosis. PMID- 12883327 TI - Involvement of the beta3 adrenoceptor in nebivolol-induced vasorelaxation in the rat aorta. AB - Nebivolol is a highly selective beta(1) adrenoceptor blocker with additional vasodilating properties. Although it has been shown that the nebivolol-induced vasorelaxation is nitric oxide (NO) and cGMP dependent, the receptor that mediates these actions remains controversial, and serotonergic as well as beta adrenergic pathways may be involved. Therefore, functional experiments investigating the receptor involved in nebivolol-induced vasorelaxation were performed in the rat aorta. Isolated aortic rings were exposed to cumulative concentrations of nebivolol. Nebivolol concentrations of 3 micromol/L and higher caused vasorelaxation, which was inhibited by the presence of the NO synthase inhibitor l-NNA (100 micromol/L), or by mechanical removal of the endothelium. Exposure of the vessel rings to the selective 5-HT(1A) antagonist NAN-190 (1 micromol/L) or the 5-HT(1/2) antagonist methysergide (1 micromol/L) did not influence nebivolol-induced vasorelaxation. Similarly, the incubation with the beta(2)-adrenoceptor antagonist butoxamine (50 micromol/L) did not prevent vasorelaxation. The selective beta(3)-adrenoceptor antagonist S-(-)-cyanopindolol (1 micromol/L), however, significantly counteracted the nebivolol-induced vasorelaxation. Furthermore, exposure of the aortic rings to cumulative concentrations of the beta(3) selective adrenoceptor agonist BRL37344 caused, like nebivolol, NO-dependent vasorelaxation that was antagonized by S-(-) cyanopindolol. The results suggest that nebivolol-induced NO-dependent vasorelaxation is, at least in part, caused by a beta(3)-adrenoceptor agonistic effect. PMID- 12883328 TI - Construction and in vitro testing of a novel fab-hirudin-based fusion protein that targets fibrin and inhibits thrombin in a factor xa-dependent manner. AB - Fibrin targeting of the thrombin inhibitor hirudin via chemical coupling is effective in vitro and in vivo. However, since chemical coupling has limitations, a recombinant approach was taken to improve the fibrin-targeting ability of hirudin. Additionally, to activate hirudin selectively at the target area and thereby limit side effects in an in vivo setting, the authors aimed to construct an inactive precursor molecule that is converted into an active thrombin inhibitor only upon cleavage by factor Xa. Using PCR, the coding region for hirudin was fused to parts of the genomic DNA of the IgG heavy chain that was cloned from the antifibrin antibody-producing hybridoma cell line 59D8. Additionally, a factor Xa recognition site was introduced between the antibody and the hirudin sequence. The fusion construct was then transfected into a heavy chain loss variant of the hybridoma cell line 59D8. After selection of stable hybridoma clones, the expressed fusion protein was evaluated for its molecular size (57 kd) and its binding ability to the fibrin-specific peptide Bbeta 15-22. The cleavage of the fusion protein by factor Xa was demonstrated by HPLC. The recombinant anticoagulant revealed antithrombin activity only after cleavage by factor Xa. Thus, the newly designed hirudin fusion protein revealed the anticipated functions in vitro. Further experiments are needed to prove whether this precursor anticoagulant allows a highly clot-specific and efficient thrombin inhibition in vivo. PMID- 12883329 TI - Cardiovascular effects of acute oxygen administration in healthy adults. AB - Supplementary oxygen is commonly administered in current medical practice. However, attention has recently been drawn to the potentially disadvantageous hemodynamic consequences in certain patients. Possible mechanisms underlying the cardiovascular responses to acute hyperoxia are unclear. The effects of acute oxygen administration on heart rate, blood pressure, cardiac output, systemic vascular resistance, and baroreflex sensitivity were studied in a series of randomised, placebo-controlled studies in healthy individuals, using validated, non-invasive techniques. The effects of oxygen administration on forearm blood flow responses to locally administered acetylcholine, an endothelium-dependent vasodilator, sodium nitroprusside, an endothelium-independent vasodilator, and l NG-monomethylarginine, a nitric oxide synthase inhibitor, were studied using venous occlusion plethysmography. Oxygen administration for 1 hour caused a reduction in heart rate (P < 0.01) and cardiac index (P < 0.05), and an increase in mean arterial pressure (P < 0.01), systemic vascular resistance (P < 0.05), large artery stiffness (P < 0.05), and baroreflex sensitivity (P < 0.05). There were no effects on vascular responses in the isolated forearm bed. These findings indicate that oxygen administration causes acute effects on cardiovascular function, which might be important in the context of acute illness. PMID- 12883330 TI - Effect of atorvastatin on ApoE and ApoC-I synthesis and secretion by THP-1 macrophages. AB - Apolipoprotein (apo) E and C-I are plasma apolipoproteins that have been implicated in the etiology of atherosclerosis and obesity, respectively. Both proteins are synthesized and secreted by macrophages, though pharmacological regulation of their production is poorly understood. The authors compared the effect of 2 HMG-CoA reductase inhibitors, atorvastatin and cerivastatin, on the synthesis and secretion of apoE and apoC-I by THP-1 macrophages. Atorvastatin reduced medium apoE and cellular apoE mRNA of PMA-activated THP-1 cells in a dose dependent manner (-24% and -22%, respectively, at 1-micromol/L, P < 0.01). ApoC-I in the medium was also reduced by atorvastatin in a dose-dependent manner, though to a lesser extent (-15% at 1-micromol/L, P < 0.05). Cerivastatin similarly reduced medium apoE (-20% at 1-micromol/L, P < 0.05) and cellular apoE mRNA (-31% at 1-micromol/L, P < 0.05), and significantly lowered cellular apoC-I mRNA (-15%, P < 0.05), but not apoC-I in the medium. In experiments with THP-1 macrophages loaded with cholesterol (ie, 24-hour incubation with acetyl-LDL), atorvastatin and cerivastatin (1-micromol/L) significantly (P < 0.05) reduced both medium apoE (-30% and -25%, respectively) and cellular apoE mRNA (-25% and -17%, respectively). A lower and less consistent effect was observed on medium apoC-I ( 6% and -18%, respectively) and cellular apoC-I mRNA (-13% and -19%, respectively). These data demonstrate that statins have the capacity to reduce the synthesis and secretion of both apoE and apoC-I in THP-1 macrophages loaded or unloaded with cholesterol. PMID- 12883331 TI - Intracellular redox status modulates monocyte chemoattractant protein-1 expression stimulated by homocysteine in endothelial cells. AB - Homocystinemia has been identified as an independent risk factor for atherosclerosis. Monocyte chemoattractant protein-l (MCP-l) is a potent chemokine that stimulates the migration of monocytes into the intima of the arterial wall. The authors investigated the role of intracellular redox status in the expression of MCP-l stimulated by homocysteine in endothelial cells. Homocysteine stimulated MCP-1 mRNA expression and protein production in a time-dependent and dose dependent manner in endothelial cells, decreased intracellular glutathione (GSH) and protein thiol levels, as well as G6PDH activity and NADPH levels. Thiol reduced reagents, GSH, and dithiothreitol levels, and reversed the MCP-l mRNA expression and protein production in endothelial cells; in addition, thiol oxidized reagent, diamide, and BSO levels, and markedly potentiated homocysteine mediated up-regulation of MCP-l mRNA expression and protein production in endothelial cells. These results demonstrate that homocysteine can trigger overexpression of the MCP-1 gene by altering the intracellular redox status, suggesting that the homocysteine-induced changes in the intracellular redox status play an important role in modulating the expression of MCP-l in endothelial cells. PMID- 12883332 TI - Cross-talk between beta-adrenergic stimulation and estrogen receptors: isoproterenol inhibits 17beta-estradiol-induced gene transcription in A7r5 cells. AB - The hypothesis that cross-talk between membrane-active beta-adrenergic agonists and estrogens includes beta-adrenergic modulation of estrogen receptor (ER) regulated gene expression was investigated. Vascular smooth muscle-derived A7r5 cells were transfected with an ERalpha expression plasmid (pCR3.1-hERalpha), the estrogen response element (ERE)-linked reporter pERE-E1b-luciferase (ERE-Luc), and pCMV-beta-galactosidase using a lysine-conjugated adenovirus transfection method. Hormone or agonist treatment and harvest followed 6 hours and 24 hours later, respectively. Treatment with 17beta-estradiol (E(2), 1 nmol/L) significantly stimulated ERE-Luc activity. Isoproterenol (10-9 to 10-6 mol/L) treatment alone did not stimulate ERE-Luc activity. Cotreatment with both E(2) and isoproterenol resulted in complete inhibition of E(2)-stimulated ERE-Luc activity. This isoproterenol effect was prevented by the beta-adrenergic antagonist propanolol (10-6 mol/L). Adrenomedullin treatment in these cells (1-50 nmol/L) did not inhibit ER/ERE-Luc activity, whether in the presence or absence of E(2). Moreover, isoproterenol did not affect vitamin D-stimulated VDRE-Luc expression, indicating that the inhibitory effect of isoproterenol on E(2) directed ERE-Luc expression is specific among nuclear transcription factor receptors. Moreover, in MCF-7 breast cancer cells, there was no effect of isoproterenol on ER/ERE-directed transcription in the absence or presence of E(2), demonstrating tissue specificity of this isoproterenol effect. These studies demonstrate cross-talk between the beta-adrenergic agonist isoproterenol and ER-directed reporter gene expression in A7r5 cells. Furthermore, this cross talk is specific with respect to agonist, nuclear receptor species, and cell type. These observations may have important implications both for the use of beta adrenergic agents to treat hypertension and for possible gender-related differences in cardiovascular regulation. PMID- 12883333 TI - Specific alterations of endothelial signal transduction pathways of porcine epicardial coronary arteries in left ventricular hypertrophy. AB - Coronary endothelial dysfunction in left ventricular hypertrophy (LVH) can reduce myocardial perfusion and result in an impaired global LV function. The objective of this study was to characterize the specific alterations of endothelial signal transduction of coronary arteries in a swine LVH model. Aortic banding was performed 3 cm above the coronary ostia. Vascular reactivity studies were performed to assess the nitric oxide (NO) and the EDHF-mediated relaxations. There was a significant increase in LV/body weight ratio associated with an increased in LV diastolic and end-diastolic pressure and decrease in dP/dT (P < 0.05), with no significant difference in coronary pressures 60 days after pressure-overload LVH. There was a significant decrease in endothelium-dependent relaxations to serotonin (5-HT) and to bradykinin (BK) (P < 0.05 for both) from LVH animals. There was no significant decrease of relaxations in the presence of BK and Nomega-l-arginine (EDHF pathway). Plasma NO(x) levels decreased significantly from 1.8% +/- 0.2% to 1.2% +/- 0.1% (P < 0.05 versus control). Chronic pressure-overload LVH is associated with an endothelial dysfunction involving both Gi and Gq protein-mediated relaxations in coronary arteries as well as the EDHF pathway. PMID- 12883334 TI - Hematein inhibits atherosclerosis by inhibition of reactive oxygen generation and NF-kappaB-dependent inflammatory mediators in hyperlipidemic mice. AB - Hematein, a natural compound, is a known anti-inflammatory and antiatherogenic agent in the rabbit model. The authors investigated the effects of this compound on atherogenesis and possible mechanisms of the actions in the hyperlipidemic mice. Low-density lipoprotein receptor-deficient (Ldlr-/-) mice fed a high cholesterol diet alone for 8 weeks developed the fatty streak lesion in the aortic sinus, whereas this lesion was significantly reduced by hematein treatment without a change in plasma lipid levels compared with control mice. Hematein treatment reduced plasma levels of lipid peroxide and superoxide generation in LPS-stimulated peritoneal macrophage. Hematein treatment inhibited NF-kappaB-DNA binding activity in peritoneal macrophages from Ldlr-/- mice and the activation of NF-kappaB in RAW264.7 macrophages. This compound suppressed plasma nitrite/nitrate levels in Ldlr-/- mice and NO production and iNOS expression in LPS+IFNgamma-stimulated peritoneal macrophages. Hematein treatment also suppressed the activity of iNOS promoters in RAW264.7 macrophages, and reduced the plasma levels of TNF-alpha and IL-1beta and the production of these cytokines in LPS+IFNgamma-stimulated peritoneal macrophages. These results suggest that hematein inhibits atherosclerotic lesion formation, possibly by reducing proinflammatory mediators through a decrease in reactive oxygen species generation and NF-kappaB activation. PMID- 12883335 TI - Amlodipine improves vascular function in patients with moderate to severe hypertension. AB - The long-term effects of amlodipine, a calcium channel blocker, were examined in patients with moderate to severe hypertension. Eighteen never-treated patients with moderate to severe essential hypertension (49 +/- 8 years) were studied. In all patients, forearm blood flow (FBF) was measured by plethysmography before and 6 months after amlodipine treatment. Endothelium-nondependent and endothelium dependent vasodilations were assessed by intrabrachial infusion of sodium nitroprusside (SNP) and acetylcholine (ACh), respectively. FBF modification by vitamin C, an oxygen radical scavenger, was also assessed under ACh infusion. The results were compared with those of 13 normal subjects. Blood pressure was significantly lowered (from 176 +/- 17/97 +/- 13 to 144 +/- 12/82 +/- 10 mm Hg) after treatment (P < 0.01). Forearm vascular resistance was increased in the hypertensive patients before treatment; however, it was normalized after treatment during SNP infusion and was improved during ACh infusion. Since vitamin C improved FBF under ACh infusion both before and after the amlodipine treatment, it is suggested that the production of free radicals was not canceled by amlodipine. The analysis of heart rate variability showed that amlodipine does not activate sympathetic nerve function. Therefore, amlodipine is effective in lowering blood pressure associated with the improvement of vascular function, and is suggested to be an effective antihypertensive agent for patients with moderate to severe hypertension. PMID- 12883336 TI - Effect of atorvastatin on the concentration, relative distribution, and chemical composition of lipoprotein subfractions in patients with dyslipidemias of type IIA and IIB. AB - The authors investigated the effect of atorvastatin (40 mg qd) on low-density lipoprotein (LDL) particle distribution in patients with dyslipidemias of type IIA (n = 55) and IIB (n = 21). Atorvastatin therapy induced a significant decrease in total and LDL cholesterol in both patient groups. A significant reduction in triglyceride values, which was more profound in type IIB patients, was also observed. In type IIA patients, LDL-3 was the predominant subfraction. Atorvastatin therapy induced a significant reduction in total LDL mass in this group of patients that was mainly due to the reduction in large and intermediate subspecies (LDL-1 to LDL-3), whereas the mass of dense LDL particles (LDL-4 and LDL-5) remained unchanged. As a consequence, the percentage contribution of dense subfractions to the total LDL mass increased significantly after atorvastatin therapy. The dense LDL-4 subfraction was the predominant one in type IIB patients. In this group, atorvastatin therapy resulted in a significant reduction in the total LDL mass, which was due to the reduction in all LDL subfractions. Thus, the percentage mass distribution of LDL particles remained unaffected. These results suggest that the effect of atorvastatin on LDL subfractions is affected by the underlying genetic defect. PMID- 12883337 TI - Endothelial release of tissue-type plasminogen activator in the human forearm: role of nitric oxide. AB - Release of tissue-type plasminogen activator (t-PA) from the vascular endothelium is paramount to endogenous thrombolysis potential. In addition to its vasodilator effects, nitric oxide (NO) has important antithrombotic properties, such as inhibition of platelet aggregation. It is currently not clear whether NO influences the capacity of the endothelium to release t-PA. The authors determined whether net endothelial t-PA release is regulated, at least in part, by NO. Endothelial t-PA release was determined, in vivo, in response to intrabrachial infusions of bradykinin (12.5-50.0 ng.100 mL tissue-1.min-1) in the presence and absence of the NO synthase inhibitor, NG-monomethyl-l-arginine (l NMMA; 5 mg/min) in 12 healthy men. Net release of t-PA across the forearm vascular bed was calculated as the product of arteriovenous concentration gradient and forearm plasma flow. The vasodilator response to bradykinin was significantly blunted ( approximately 30%) with l-NMMA. Although there was no effect of l-NMMA on basal t-PA release, acute release of t-PA to bradykinin was higher (P < 0.01) after (from -0.2 +/- 0.5 to 105.2 +/- 9.4 ng.100 mL tissue 1.min-1) versus before (from -0.4 +/- 0.7 to 48.7 +/- 7.3 ng.100 mL tissue-1.min 1) the administration of l-NMMA. Thus, in the absence of NO endothelial t-PA release was enhanced. These results suggest a potential regulatory influence of NO on bradykinin induced endothelial t-PA release. PMID- 12883338 TI - Melting after keratoprosthesis implantation: the effects of medroxyprogesterone. AB - PURPOSE: This study was conducted to evaluate the effect of topical medroxyprogesterone (MPG) following KPro implantation in human subjects in whom donor tissue grafts had been contraindicated by high risk of failure. METHODS: Outcomes of implantation of the Chirila KPro, now known as AlphaCor, were reviewed with respect to postoperative MPG therapy. Ten of 45 (22%) patients had received MPG for a period of 12 months, while 35/45 (78%) had not. MPG treatment was halted because the drug is not approved as an adjunctive treatment of KPro patients. The main outcome measures were the incidence and timing of corneal stromal melting and visual acuity. RESULTS: Of those untreated with MPG, 34% developed a melt (mean follow-up 9.7 months), whereas of those who received MPG, 60% developed a melt (mean follow-up 28.4 months). However, mean time to melt onset for untreated cases was 8.8 months, whereas mean time to melt onset for treated cases was 23.2 months. There is a statistically significant association between time to melt onset, where melts occurred, and MPG therapy (chi2 = 0.001). In both groups, melts were strongly associated with a history of ocular HSV, which represented 17.1% of untreated and 20% of treated cases and is now considered a contraindication for AlphaCor. Preoperative visual acuities were in the range Perception Light (PL)-Count Fingers (CF) in all cases, whereas mean best postoperative best corrected visual acuity was 20/200 (range PL-20/30) in untreated cases and was 20/120 [range Hand Movements (HM)-20/30)] in MPG-treated cases. CONCLUSIONS: Although MPG may not influence the underlying incidence of melt-related complications, which are likely to be associated with other risk factors especially HSV, it may have a protective effect with regard to melt onset and severity. Controlled studies would assist evaluation of its use in this indication. PMID- 12883339 TI - Outcome of double-headed pterygium surgery. AB - PURPOSE: To evaluate the postoperative outcome and recurrence rate of bare sclera technique combined with intraoperative application of mitomycin C (MMC) 0.02% for 5 minutes in the treatment of primary and recurrent double-head pterygia. METHODS: A prospective, noncomparative interventional case series of 13 eyes in 13 patients with primary (n = 10) or recurrent double-head pterygia in one eye operated on by one surgeon (R.A.). They all had bare sclera excision combined with intraoperative application of MMC 0.02% for 5 minutes. All patients were followed postoperatively for recurrence of the lesion, aesthetic outcome, and incidence of eye complications. RESULTS: Mean follow-up was 36.3 +/- 3.8 months for the patients with primary and 28.4 +/- 2.7 months for the patients with recurrent double-head pterygia. There was only one recurrence in one of the three eyes in the group with recurrent pterygia (33.33%). The only significant complication observed was a pyogenic granuloma in two of the recurrent and one of the primary cases. CONCLUSIONS: The bare sclera technique combined with intraoperative MMC 0.02% for 5 minutes is an effective and safe procedure for double-head pterygia. This method can serve as a useful alternative to amniotic membrane transplantation in countries where the latter is not available and in patients in whom conjunctival autograft is not feasible. PMID- 12883340 TI - Accuracy and precision of the amadeus microkeratome in producing LASIK flaps. AB - PURPOSE: To evaluate the accuracy and precision of corneal flap thickness following laser in situ keratomileusis (LASIK) performed using the 140-, 160-, and 180-microm heads for the Amadeus microkeratome (AMO, Irvine, CA). SETTING: The study took place at the Cullen Eye Institute, Baylor College of Medicine, Houston. METHODS: In this prospective study, using the Amadeus microkeratome, LASIK flaps were cut in 51 right eyes and 50 left eyes with the 140-microm head, 25 right eyes and 25 left eyes with the 160-microm head, and five right eyes and one left eye with the 180-microm head. The same microkeratome blade was used for bilateral cases with the right eyes always undergoing surgery first. Eyes were grouped by order of blade use for statistical analysis. The effect of preoperative corneal thickness, keratometry values, blade oscillation and translation speeds, and blade reuse on flap thickness was evaluated. RESULTS: Mean flap thicknesses were 153 +/- 18 (range 97-187 microm) OD and 134 +/- 25 microm (range 79-174 microm) OS for the 140-microm head; 182 +/- 26 microm (range 105-220 microm) OD and 163 +/- 29 microm (range 105-216 microm) OS for the 160 microm head; and 235 +/- 24 microm (range 198-258 microm) for the 180-microm head. Flap thickness was significantly thicker for the first eyes cut (right eyes) and was positively correlated with increasing corneal thickness in both eyes. For the first eyes cut, flap thickness was also significantly thicker than the labeled thickness specified by the manufacturer. CONCLUSIONS: With the Amadeus microkeratome, LASIK flap thickness correlated with central corneal thickness for the 140-microm head. Reuse of the microkeratome blades produced significantly thinner LASIK flaps on second eyes cut. PMID- 12883341 TI - Identification of novel mutations of the CHST6 gene in Vietnamese families affected with macular corneal dystrophy in two generations. AB - PURPOSE: To report the clinical and genetic findings of Vietnamese families affected with macular corneal dystrophy (MCD) in 2 generations. METHODS: Two families, including 7 patients and 3 unaffected members, were examined clinically. Blood samples were collected. Fifty normal Vietnamese individuals were used as controls. Genomic DNA was extracted from leukocytes. Analysis of the carbohydrate sulfotransferase (CHST6) gene was performed using polymerase chain reaction and direct sequencing. RESULTS: The typical form of MCD was recognized in family B, in which sequencing of CHST6 gene revealed an nt 1067 1068ins(GGCCGTG) mutation (frameshift after 125V) homozygously in MCD patients and heterozygously in the unaffected members. Family N also showed clinical features of MCD, moderate in the mother but severe in the affected son. Sequencing revealed a single heterozygous Arg211Gln in the mother, compound heterozygous Arg211Gln+ Gln82Stop in the affected son, and heterozygous Arg211Gln mutation in the unaffected members. The identified mutations in these pedigrees were excluded from normal controls. CONCLUSIONS: The novel frameshift and compound heterozygous mutations might be responsible for MCD in the families studied. The phenotypic variation between affected parents and offspring was unclear. In family N, severe MCD phenotype seen in the affected son may be due the fact that he had an early stop codon mutation (Gln82Stop). PMID- 12883342 TI - Comparison of confocal biomicroscopy and noncontact specular microscopy for evaluation of the corneal endothelium. AB - PURPOSE: To compare the clinical efficacy of confocal biomicroscopy with that of noncontact specular microscopy for the evaluation of the corneal endothelium. METHODS: The corneal endothelium was examined in 14 normal subjects (28 eyes) and in 6 patients (11 eyes) with Fuchs corneal endothelial dystrophy using a noncontact specular microscope (SP-2000P, Topcon, Japan) and a confocal biomicroscope (ConfoScan, Tomey, Japan). The images and the calculated densities of corneal endothelial cells obtained by the 2 techniques were compared. RESULTS: For normal subjects, the images of corneal endothelial cells obtained by the 2 techniques were almost identical, although the density of these cells determined by confocal biomicroscopy (2916 +/- 334 cells/mm2) was slightly higher than that determined by noncontact specular microscopy (2765 +/- 323 cells/mm2). In contrast, whereas clear images of corneal endothelial cells, allowing the determination of cell density, were obtained for all 11 eyes of the patient group by confocal biomicroscopy, clear images were obtained for only 4 of these 11 eyes (36.4%) by noncontact specular microscopy. CONCLUSION: Both noncontact specular microscopy and confocal biomicroscopy revealed the shapes and number of endothelial cells in the normal cornea. However, for corneas with Fuchs dystrophy, clear images were obtained only by confocal biomicroscopy. Confocal biomicroscopy is thus an effective tool for evaluation of the diseased corneal endothelium. PMID- 12883343 TI - Is complete androgen insensitivity syndrome associated with alterations in the meibomian gland and ocular surface? AB - PURPOSE: This study's purpose was to determine whether complete androgen insensitivity syndrome (CAIS) is associated with alterations in the meibomian gland and ocular surface. METHODS: Individuals with CAIS, as well as age-matched female and male controls, completed questionnaires which assessed dry eye symptoms and underwent slit lamp evaluations of the tear film, tear meniscus, lids and lid margins and conjunctiva. The quality of meibomian gland secretions was also analyzed. RESULTS: Our results demonstrate that CAIS is associated with meibomian gland alterations and a significant increase in dry eye signs and symptoms. Clinical assessment revealed that CAIS women, as compared to controls, had a significant increase in telangiectasia, keratinization, lid erythema and orifice metaplasia of the meibomian glands, and a significant decrease in the tear meniscus and quality of meibomian gland secretions. Questionnaire results showed that dry eye symptoms were increased over twofold in CAIS individuals, as compared to controls, including a significant increase in the sensations of dryness, pain and light sensitivity. CONCLUSION: Our results suggest that androgen insensitivity may promote meibomian gland dysfunction and an increase in the signs and symptoms of dry eye. PMID- 12883344 TI - 5-Fluorouracil as chemoadjuvant for primary pterygium surgery: preliminary report. AB - PURPOSE: To investigate the safety and efficacy of intraoperative application of 5-fluorouracil as an adjuvant in primary pterygium surgery and to evaluate the effect of postoperative subconjunctival 5-fluorouracil injections on the recurrent pterygium. METHODS: Of 25 consecutive white patients, 28 eyes with primary pterygium underwent pterygium excision with intraoperative application of 5-fluorouracil (25 mg/mL for 3 minutes). The superior and inferior conjunctiva was approximated to cover the scleral bed within 1 mm of the limbus. Recurrence of pterygium was defined as postoperative fibrovascular growth more than 1 mm onto the cornea. Eyes with recurrence less than 2 mm were treated with subconjunctival 5-fluorouracil injections. RESULTS: After a mean follow-up of 14.1 +/- 3.9 months (mean +/- standard deviation), 7 recurrences (25%) were observed. All recurrences were detected within 12 months. In 4 of 7 recurrences, the fibrovascular growths were less than 2 mm. We, therefore, performed subconjunctival 5-fluorouracil injections. In 3 (75%) of 4 recurrences, the fibrovascular growths became atrophic. No serious complications were observed during and after the surgery. However, superficial punctate keratitis, pain, and hyperemia were detected in all patients in the early postoperative period. As a result, of 28 eyes, 4 (14%) had unacceptable cosmetic results and growing recurrences. CONCLUSIONS: This study suggests that intraoperative applications of 5-fluorouracil is both efficient and safe in the treatment of primary pterygium. Additionally, postoperative subconjunctival 5-fluorouracil injections may prevent the progression of fibrovascular tissue. PMID- 12883345 TI - Corneal thickness values before and after oxybuprocaine 0.4% eye drops. AB - PURPOSE: To determine changes in corneal thickness after topical anesthesia. METHODS: Corneal thickness was measured before and 3 minutes after administration of two drops of oxybuprocaine 0.4% to 26 patients (26 eyes). We analyzed the corneal thickness of a control group, which was made up of 26 patients (26 eyes) before and 3 minutes after administration of two drops of saline solution. Corneal thickness was measured with the Orbscan Topography System II (Bausch Lomb Surg., Barcelona). RESULTS: Variations higher than +/- 10 microm were found following the instillation of 2 oxybuprocaine eye drops in eight eyes (30.76%) at the inferonasal cornea, in six eyes (23.08%) at the superotemporal, temporal and inferotemporal cornea, in five eyes (19.23%) at the nasal cornea, in three eyes (11.53%) at the central cornea, and in two eyes (7.69%) at the superonasal cornea. Nevertheless, no significant differences in the mean corneal thickness at each corneal location between the first and the second corneal thickness measurements were found in anesthetized eyes. CONCLUSIONS: Some individuals can present important increases and decreases in corneal thickness values after anesthetic eye drops. This effect of anesthetic eye drops must be considered by refractive surgeons when carrying out preoperative laser in situ keratomileusis corneal thickness measurements. PMID- 12883346 TI - Analysis of limbal stem cell deficiency by corneal impression cytology. AB - PURPOSE: The impaired function of corneal epithelial stem cells, located in the limbus, is responsible for corneal surface damage and is clinically characterized by recurrent epithelial defects, conjunctivalization, neovascularization, and corneal opacity. The aim of this study was to investigate corneal limbal stem cell deficiency (LSCD) by means of the impression cytology (IC) technique, using antibodies against cytokeratin 19 (CK19) and cytokeratin 3 (CK3), and to evaluate the diagnostic potential of this approach. METHODS: Over a 3-year period (October 1998-June 2001), we collected 113 pairs of IC samples from the eyes of 85 patients with a range of ocular surface diseases and performed an immunocytochemical analysis of CK19 and CK3. Samples with more than 50% cellularity were considered suitable for diagnostic purposes, while samples with less than 50% cellularity were considered with caution. CK19-positive cells in corneal IC were considered an expression of LSCD. We arbitrarily scored LSCD as mild (<25% of CK19-positive cells), moderate (25-50%), and severe (>50%). RESULTS: One hundred thirteen pairs of IC specimens were obtained from 85 patients; 32 patients (37.6%) had alkaline burns, 18 (21.2%) had other chemical or physical corneal injuries, 13 (15.3%) had complications from wearing contact lenses, 8 (9.4%) had severe microbial keratitis, and 14 (16.5%) had suspicious limbal deficit due to other causes. Nine patients underwent bilateral sampling and 12 had to be resampled. Thirteen pairs of IC specimens were obtained during the follow-up of 8 patients who had undergone limbal stem cell transplantation. In 3 of these patients, IC confirmed reversion to corneal immunophenotype (CK3+/CK19-), whereas in 4, residual limbal damage was still evident; 1 patient relapsed. In the remaining 100 pairs of impressions, we found 77 cases of LSCD, whereas in 16 pairs, we did not find LSCD. Seven pairs were defined as "not valuable" because of the poor quality of both CK samples. Diffuse LSCD, moderate or severe in degree, was found in 26 of 32 patients (81.2%) with alkali burns, whereas mild diffuse LSCD or sectoral LSCD was found in 13 of 18 patients (72.2%) with other chemical-physical injuries, in 10 of 13 patients (76.9%) wearing contact lenses, in 7 of 8 patients (87.5%) with severe microbial keratitis, and in 12 of 14 patients (85.7%) with other corneal pathologies. The quality of impressions was assessed in 77 cases and found to be good or discrete for both CKs in 32 cases (41.5%) and poor in 45 (58.5%): in 46.7% of these cases, the IC was poor only for CK19 and in 45.4% only for CK3. CONCLUSIONS: Immunocytochemistry for seeking out CK19- and CK3-positive cells on corneal IC is a simple and practical method to investigate LSCD. We believe that this technique could have an important role in evaluating patients undergoing therapeutic penetrating keratoplasty to select those who would benefit from limbal stem cell transplantation. Since sampling has been shown to be a critical point, we believe that any improvement in this area will also help to improve the methodology and will contribute to its wider utilization. PMID- 12883347 TI - The effect of eye closure on the post-lens tear film thickness during silicone hydrogel contact lens wear. AB - PURPOSE: The purpose of this study was to investigate the effect of eye closure on the thickness on the post-lens tear film (POLTF) during silicone hydrogel contact lens wear. METHODS: Ten subjects (mean age, 30.2 +/- 8.6 years; seven males) wore balafilcon A silicone hydrogel contact lenses in both eyes during the experimental procedure (power = -2.00 D, base curve = 8.6 mm). Previously described interference techniques, based on oscillations in reflectance spectra, were used to measure the post-lens tear film. Reflectance spectra (562-1030 nm) from the front of the eye wearing a contact lens were measured at normal incidence where the thickness of the POLTF was derived from the "frequency" of the oscillations. Six baseline measures of POLTF thickness were taken, followed by eye closure in the supine position for 30 minutes. Post-lens tear film thickness measures were taken after 5 and 15 minutes of eye closure. After 30 minutes of eye closure, 1 measure of POLTF was taken every minute for an additional 15 minutes. RESULTS: The average baseline POLTF thickness was 2.00 +/- 0.30 microm (median, 2.00) for the 10 subjects. The average POLTF thickness decreased to 1.58 +/- 0.33 and 1.20 +/- 0.23 microm after 5 and 15 minutes of eye closure, respectively (repeated measures ANOVA, F = 42.18, P < 0.0001). Only 5 of 60 POLTF thickness estimates were obtained for the first 6 minutes following completion of 30 minutes of eye closure, indicating that the POLTF may be too thin (i.e., <1 microm) to measure using this method. However, there was a significant increase in POLTF thickness for 15 minutes after prolonged eye closure (mixed modeling regression, thickness = 0.85 + 0.07 x time after eye opening; P < 0.0001). CONCLUSIONS: The POLTF thickness is rapidly reduced by eye closure during contact lens wear. After 30 minutes of eye closure, the POLTF thickness may be reduced to values less than 1 microm for the first several minutes after opening the eyes. PMID- 12883348 TI - Effects of minocycline on the ocular flora of patients with acne rosacea or seborrheic blepharitis. AB - PURPOSE: To assess the effect of minocycline on the ocular flora in patients with acne rosacea or blepharitis. METHODS: A total of ten patients were enrolled in this prospective study, with six patients diagnosed with acne rosacea with concomitant meibomianitis, two patients with acne rosacea without concomitant ocular involvement, and two patients with seborrheic blepharitis. The eyelids and conjunctiva of both eyes were cultured before the initiation of systemic minocycline therapy, after 3 months of active therapy, and 3 months after the discontinuation of therapy. Isolated bacteria were identified and quantified, and antibiotic susceptibility was determined. RESULTS: The colony-forming units (CFU) isolated from the eyelids significantly decreased after a 3-month treatment with minocycline (P = 0.0013). The CFU significantly increased to approach that of the baseline with the discontinuation of minocycline (P = 0.0275). The most common isolated bacteria, including coagulase-negative Staphylococcus (CNS), Staphylococcus aureus (S. aureus), and Propionibacterium acne (P. acne), except for corynebacterium, had a significant decrease in bacterial count with minocycline therapy compared with baseline (P < 0.05). There was a trend in the decrease of bacterial CFU isolated from the conjunctiva with minocycline therapy, although this was not statistically significant (P = 0.1955). Four of the ten patients carried tetracycline-resistant CNS strains, but none of the S. aureus or P. acne isolated at baseline was resistant to tetracycline. All six patients with acne rosacea and concomitant meibomianitis had marked clinical improvement. CONCLUSION: Minocycline effectively decreased eyelid bacterial flora in patients with acne rosacea or blepharitis. One of the mechanisms of newer generation tetracycline analogues may be a decrease or elimination of bacterial flora from the eyelids. PMID- 12883349 TI - Technique of area measurement of epithelial defects. AB - PURPOSE: To compare two commonly used methods of area measurement of corneal epithelial defects, namely, estimation of the area of an equivalent rectangle and use of image analysis software, the gold standard. METHODS: Thirty-eight patients with corneal epithelial defects were evaluated. The largest linear dimension of the epithelial defect and its largest possible perpendicular within the confines of the epithelial defect were measured using a slit lamp. The area was then measured using digital photographs and image analysis software. A paired sample t test was used to compare the results. RESULTS: The two measurements of area correlated well (P = 0.84); their difference was insignificant (P = 0.063) and a linear regression relation between the two could be derived with an r2 value of 0.71, which was significant. CONCLUSIONS: Accurate slit-lamp measurements with estimation of the area of an equivalent rectangle can safely be used as a measure of the epithelial defect size when sophisticated image analysis software is not available. PMID- 12883350 TI - Long-term outcome in high-risk corneal transplantation and the influence of HLA-A and HLA-B matching. AB - PURPOSE: To evaluate long-term follow-up of high-risk corneal transplants allocated after matching for broad HLA-A and HLA-B antigens and to establish whether matching for HLA-A and -B antigen "splits" would result in a reduced risk of immunologic graft failure. METHODS: A total of 303 high risk corneal transplants was included. Class I antigen-matched donor corneas were obtained using broad HLA-A and -B antigen data and accepting 0 or 1 mismatch at each locus. Analysis of HLA antigens was performed also on the split typing level. The influence on immunologic graft failure for an increasing number of matched class I antigens based on split typing was analyzed with Kaplan-Meier statistics and Cox regression. Graft survival and indication for transplantation were investigated. RESULTS: Rejection was the cause of 34% of all graft failures. A significantly higher immune failure free graft survival was found in a group with 0 or 1 HLA-A and -B mismatch based on split typing (log-rank test, P = 0.002). A beneficial effect of matching for split antigens was shown with multivariate analysis (odds ratio, 0.41). CONCLUSIONS: One third of graft failures in our high risk population was caused by irreversible graft rejection. Allocation of donor corneas based on a 0 or 1 split antigen mismatch at both HLA-A and -B loci could contribute to a higher immune failure free graft survival and could result in a higher overall graft survival. PMID- 12883351 TI - Synergistic effect of TNF-alpha and either IL-4 or IL-13 on VCAM-1 expression by cultured human corneal fibroblasts. AB - PURPOSE: To examine the role of corneal fibroblasts in the pathogenesis of vernal keratoconjunctivitis, we investigated the effects of tumor necrosis factor (TNF) alpha, interleukin (IL)-4, and IL-13 on the expression of vascular cell adhesion molecule (VCAM)-1 by cultured human corneal fibroblasts. METHODS: Cultured human corneal fibroblasts were incubated with various combinations and concentrations of TNF-alpha, IL-4, and IL-13. The cell surface expression of VCAM-1 was subsequently evaluated by whole-cell enzyme-linked immunosorbent assay and immunocytochemistry, and the abundance of VCAM-1 mRNA in cell lysates was determined by quantitative reverse transcription and polymerase chain reaction analysis. RESULTS: Corneal fibroblasts incubated in the absence of cytokines exhibited minimal expression of VCAM-1. Whereas incubation of the cells with TNF alpha, IL-4, or IL-13 alone, or with the combination of IL-4 and IL-13, induced only a small increase in VCAM-1 expression, exposure of the cells to TNF-alpha in combination with either IL-4 or IL-13 resulted in a marked synergistic increase in expression of this adhesion molecule that was both time and dose dependent. The abundance of VCAM-1 mRNA in corneal fibroblasts was also increased in a synergistic manner by incubation of the cells with TNF-alpha together with either IL-4 or IL-13. CONCLUSION: Stimulation of human corneal fibroblasts with the combination of TNF-alpha and either IL-4 or IL-13 resulted in synergistic increases in both the abundance of VCAM-1 mRNA and the cell surface expression of VCAM-1 protein. This cytokine-induced increase in VCAM-1 expression by corneal fibroblasts may contribute to eosinophil infiltration in corneal lesions associated with vernal keratoconjunctivitis. PMID- 12883352 TI - Q-Switched 2.94-microm Er:YAG laser trephination with convergent and divergent cut angles for penetrating keratoplasty. AB - PURPOSE: To study the morphologic properties of perpendicular (P), convergent (C), and divergent (D) cut angles using different speeds of rotations during donor and recipient nonmechanical trephination for experimental penetrating keratoplasty. METHODS: With a Q-switched 2.94-microm Er:YAG laser corneal trephination was performed in 150 enucleated porcine eyes using ceramic open masks with 8 "orientation teeth/notches" and an automated globe rotation device allowing different cut angles [0 degree (P), 10 and 20 degrees (C and D)] toward the optical axis and variation of the rotation speed [3, 7, and 11 rotations per minute (rpm)]. The regularity of the cut (I, regular; II, slightly irregular; III, irregular) was assessed by light microscopy. The area of thermal damage and the number and size of "spikes" in the stroma at the superficial, intermediate and deep level of the excision were analyzed using digital images and the Optimas image processing software. RESULTS: The regularity of the cut was classified as I in 42%/22% of donor/recipient and as II in 41%/56%, respectively. The thermal damage was least expressed with D20 degree cut angle and donor mask (P < 0.01). With all cut angles and speeds of rotation, thermal damage at the deep level of excision was significantly smaller (P < 0.01). With different speeds of donor rotations, the thermal damage showed no significant difference. With recipient trephination, the thermal damage at the deep level was greatest with 7 rpm (P < 0.01). The number and size of spikes of thermal damage with donor and recipient masks were significantly smaller in the deep stroma (P < 0.01). CONCLUSIONS: Q switched Er:YAG laser trephination with appropriate settings results in low thermal damage zones at the cut margin. Different cut angles and speeds of trephination may affect the cut performance and quality of the excision. In our study, low rotation speed and divergent donor cut angles showed the best results. The cut quality and the small thermal damage with the Q-switched 2.94-microm Er:YAG laser seem to be tolerable for corneal trephination. Therefore, this modality may be a low-cost, easy-to-handle alternative for nonmechanical corneal transplantation in humans. PMID- 12883353 TI - How long can donor sclera be safely stored? AB - PURPOSE: To determine whether after prolonged storage of sclera in glycerin, there is any bacteriologic contamination that will reactivate, whether reconstituted sclera retains its tensile strength, and whether sclera retains its microstructural integrity. METHODS: Sixty-six scleral shells stored in glycerin for 9 to 19 years, as well as 11 controls stored for 6 months to 4 years, were studied by cutting a small wedge of tissue from the anterior margin of each and directly inoculating into thioglycolate broth, cutting an equatorial ring and determining its break strength using a tensiometer, and cutting a small piece from the remaining posterior portion and examining by scanning electron microscopy. RESULTS: After such prolonged storage, bacteriologic contamination was not detected, tensile strength generally increased with increasing duration of storage, and ultrastructural integrity was maintained on scanning electron microscopy. CONCLUSIONS: This study suggests that storage of scleral shells can be safely prolonged; we hope this can facilitate an increased supply of donated sclera to patients and surgeons. PMID- 12883354 TI - Confocal microscopy in ocular chrysiasis. AB - PURPOSE: To describe the presence of gold salt deposits in the corneal tissue of a patient treated with gold sodium thiomalate through confocal microscopy and to relate our findings to temporary corneal dynamics. METHODS: A patient who had been treated with gold sodium thiomalate for 6 years was referred from the Rheumatology Unit of our center for an ophthalmologic examination. At the time of examination she had been treatment-free for 9 years. Besides visual acuity, the examination included slit-lamp biomicroscopy, intraocular pressure, and fundoscopy. Confocal microscopy performed using a Confoscan P4 instrument with a 40x lens to obtain 15 images per second on an S-VHS video system (Panasonic) for subsequent analysis. RESULTS: The basic layered structure of the cornea was unaffected, but the presence of highly reflective particles was noted. These particles were interpreted as gold deposits in the corneal stroma, especially in the anterior and middle stroma. CONCLUSION: Confocal microscopy is a useful method of evaluating the deposition of gold salts in tissues, particularly the cornea. Its future use will help gain a deeper understanding of corneal metabolism. PMID- 12883355 TI - Acanthamoeba as a cause of peripheral ulcerative keratitis. AB - PURPOSE: To report a case of peripheral ulcerative keratitis caused by Acanthamoeba. METHODS: Case report and review of the literature. A 37-year-old woman with a history of pain and redness of the right eye with no apparent predisposing factors, on examination, revealed a peripheral ulcerative keratitis. RESULT: Microbiological investigations of the corneal infiltrate revealed Acanthamoeba cysts. CONCLUSION: All cases of peripheral ulcerative keratitis should be subjected to routine microbiological evaluation including those for Acanthamoeba. PMID- 12883356 TI - In vivo confocal microscopy in the patients with cornea farinata. AB - PURPOSE: To report in vivo corneal confocal microscopic findings of patients with cornea farinata. PATIENTS AND METHODS: Two unrelated patients, a 47-year-old man and a 77-year-old woman, with cornea farinata were studied. Examination with a confocal microscope was performed in addition to routine slit-lamp biomicroscopy. RESULTS: In both cases, slit-lamp biomicroscopy showed numerous small, faint opacities in the deep stroma in both eyes. Using confocal microscopy, highly reflective small particles were observed in the cytoplasm of keratocytes in the deep stroma adjacent to the corneal endothelial layer. No abnormalities could be detected in the epithelial layer, in the mid-stromal layer, at the level of Descemet's membrane, and in the endothelial layer. CONCLUSIONS: In vivo corneal confocal microscopy is useful for observing stromal abnormalities in cornea farinata. Further investigation of posterior stromal opacities using confocal microscopy may be useful to understand and differentiate various corneal conditions involving primarily deep stromal layers. PMID- 12883357 TI - Soft contact lens wear in identical twins. PMID- 12883358 TI - CD44s adhesive function spontaneous and PMA-inducible CD44 cleavage are regulated at post-translational level in cells of melanocytic lineage. AB - Adhesion between the CD44s receptor and hyaluronic acid plays an important role in cell migration, tumour growth and progression. Although the alternative splicing of CD44 variant exons represents the principal regulatory mechanism of CD44-mediated functions, CD44v spliced variants are scantily expressed in melanoma cells. For this reason, we have investigated the possibility that post translational modifications of the CD44 standard receptor could play a pivotal role in regulating CD44-mediated functions in melanoma. Using metabolic inhibitors of N- and O-glycosylation, as well as melanoma transfectants expressing CD44s O-glycosylation site-specific mutants, we performed structural and functional analysis of N- and O-deglycosylated CD44s molecules expressed in melanoma cells. We discovered that complete N- and O-glycosylation is not required by CD44s to be correctly expressed on the melanoma cell surface. Indeed, variably glycosylated and functionally different CD44s molecules were constitutively expressed in primary and metastatic lesions. Furthermore, we observed that changes in N- and O-glycosylation of CD44s could modulate its cleavage. In fact, spontaneous CD44s shedding was dependent on the presence of partial or complete O-glycosylation of four serine-glycine motifs localized in the membrane-proximal CD44 ectodomain. Mutation of these serine residues, as well as an extensive metabolic O-deglycosylation, strongly impaired spontaneous CD44 shedding. Furthermore, an O-glycosylation-independent mechanism of CD44 cleavage has been identified. This alternative mechanism of receptor cleavage is phorbol 12-myristate-13-acetate (PMA) inducible, mediated by metalloproteinase and requires the presence of N-linked sugar residues. Our findings demonstrate that the post-translational modification of CD44s represents the principal regulatory mechanism of CD44s-mediated functions in melanoma. PMID- 12883359 TI - Antiapoptotic action of 1alpha,25-dihydroxyvitamin D3 in primary human melanocytes. AB - 1alpha,25-Dihydroxyvitamin D(3) [1,25-(OH)(2)D(3)] has been shown to induce cell growth arrest and to possess differentiation-inducing behaviour in both primary melanocytes and melanoma cell lines. Moreover, in several melanoma cell lines it has been demonstrated that the antiproliferative action is accompanied by an increase in apoptosis. In contrast, here we show that physiological concentrations of 1,25-(OH)(2)D(3) did not induce apoptosis in primary melanocytes despite a cell growth inhibitory effect. Furthermore, treatment with 1,25-(OH)(2)D(3) made melanocytes resistant to several inductors of programmed cell death, including tumour necrosis factor-alpha and ultraviolet radiation. The antiapoptotic effect of 1,25-(OH)(2)D(3) was completely abolished by the addition of N,N-dimethylsphingosine, which blocks the formation of the sphingolipid degradation product sphingosine 1-phosphate (S1P), suggesting a crucial role for this sphingolipid in 1,25-(OH)(2)D(3)-mediated cytoprotection. Indeed, stimulation of melanocytes with S1P also resulted in an antiapoptotic action. In addition, S1P induced cell growth arrest of human melanocytes. This was an unexpected finding, as S1P is generally known as a potent mitogenic molecule in a variety of cells, including fibroblasts. As both 1,25-(OH)(2)D(3) and S1P have been identified to modify the Bcl-2/Bax ratio in epithelial cells, we also measured the expressions of these proteins; however, treatment of melanocytes with either 1,25-(OH)(2)D(3) or S1P did not alter the Bcl-2/Bax ratio. In conclusion, 1,25-(OH)(2)D(3) was shown to protect human melanocytes from apoptosis by formation of S1P, which is opposite to its apoptotic action in diverse melanoma cell lines. PMID- 12883360 TI - Growth inhibition of subcutaneous mouse melanoma and induction of natural killer cells by liposome-mediated interferon-beta gene therapy. AB - In this study we investigated the antitumour effect and mechanism of action of cationic liposome-mediated murine interferon-beta (IFNbeta) gene therapy in mouse B16F1 melanoma cells in vitro and in vivo. Murine IFNbeta gene transfer by cationic liposome resulted in substantial growth inhibition of B16F1 melanoma cells in culture when compared with phosphate buffered saline or recombinant murine IFNbeta treatment, or lacZ control gene transfer. Use of video-enhanced contrast-differential interference contrast (VEC-DIC) microscopy revealed that liposomes containing the murine IFNbeta gene [lip(pSV2muIFNbeta)], but not recombinant murine IFNbeta, induced dramatic morphological changes that characterize apoptosis, including bleb formation, shrinkage of cells, nuclear condensation and 'ballooning', in approximately 30% of the cells treated. Intratumoral administration of lip(pSV2muIFNbeta) resulted in a 5.5-fold reduction in the mean volume of subcutaneous melanoma lesions in syngeneic mice 15 days after treatment and eradicated the tumour in 18% of the mice treated. Immunocytochemical analysis demonstrated that a larger number of natural killer (NK) cells infiltrated the tumour following lip(pSV2muIFNbeta) treatment than in controls. In vivo depletion of NK cells using the anti-asialoGM1 antibody reduced the efficacy of lip(pSV2muIFNbeta) treatment. Taken together, our data suggest that cationic liposome-mediated IFNbeta gene therapy could be effective against melanoma by directly inducing cell death and stimulating NK cells. PMID- 12883361 TI - Comparison of phaeomelanin and its precursor 5-S-cysteinyldopa in the serum of melanoma patients. AB - 5-S-Cysteinyldopa (5-S-CD) has been used as a biochemical marker of melanoma progression. Recently we have shown that the serum level of 5-S-CD is a sensitive and specific marker in predicting distant metastases. In melanocytes and melanoma cells, cysteinyldopa isomers are oxidized to phaeomelanin, the yellow to reddish melanin pigment. In this study we have developed a new method to measure levels of phaeomelanin in serum samples and have evaluated its clinical significance. The method is based on the production of 4-amino-3-hydroxyphenylalanine (4-AHP) and 3-amino-4-hydroxyphenylalanine (3-AHP) on reductive hydrolysis of phaeomelanin with hydriodic acid. 3-AHP is also derived from 3-nitrotyrosine containing proteins. The isomeric 4-AHP and 3-AHP can be separated by high performance liquid chromatography. The mean +/- SD serum levels of 5-S-CD in control subjects (n = 36), in melanoma patients without recurrence (n = 92) and in melanoma patients with metastases (n = 24) were 2.7 +/- 1.2 nM (median 2.3 nM), 4.0 +/- 1.6 nM (median 3.8 nM) and 72 +/- 105 nM (median 35 nM), respectively. The serum levels of 4-AHP in these three groups were 45 +/- 21 nM (median 31 nM), 80 +/- 75 nM (median 53 nM) and 306 +/- 627 nM (median 133 nM), respectively. The serum levels of 4-AHP in patients with metastases (100 samples from 15 patients with progressive disease) correlated well (r = 0.887) with serum levels of 5-S-CD, while serum levels of 3-AHP did not (r = 0.240). The serum 5-S CD and 4-AHP levels were serially analysed in the 15 patients with progressive disease. In two patients (13%), serum 4-AHP levels were elevated to abnormal levels before the serum 5-S-CD levels exceeded the cut-off value of 10 nM. In five patients (33%), the serum 4-AHP levels rose concurrently with the serum 5-S CD levels. In the remaining eight patients (54%), serum 4-AHP levels were of less diagnostic value. Thus, the serum phaeomelanin level appears to be less sensitive than the serum 5-S-CD level in detecting distant metastases. PMID- 12883362 TI - Germline 657del5 mutation in the NBS1 gene in patients with malignant melanoma of the skin. AB - In this study we determined in what proportion of consecutive malignant melanoma (MM) cases the 657del5 mutation of exon 6 of the NBS1 gene can be detected and whether it is associated with the occurrence of MM. Two groups of patients were studied: a series of 80 consecutive patients with histologically confirmed MM of the skin diagnosed in the city of Szczecin, Poland, and a series of 530 consecutive individuals selected at random by family doctors from the city of Szczecin. Molecular examination included an allele-specific polymerase chain reaction assay for the NBS1 founder mutation (657del5), genomic sequencing, loss of heterozygosity analysis using CA-repeat microsatellite markers, and haplotype analysis. The NBS1 founder mutation was detected in two of the 80 (2.5%) MM cases and in three of the 530 individuals (0.6%) from the general population. The difference was not statistically significant. However, examination of tumorous DNA from the patients with MM and NBS1 mutation revealed loss of heterozygosity in both cases. Haplotype analysis revealed that allellic loss affects wild-type alleles. Breast cancer was found in second-degree relatives of both MM probands with NBS1 mutations. One of these probands was simultaneously affected with breast cancer. It seems that the 657del5 mutation of exon 6 of NBS1 gene may be responsible for the occurrence of a small proportion of MM patients, characterized by the occurrence of breast cancer among their relatives. PMID- 12883363 TI - Immunoproteasome subunits LMP2 and LMP7 downregulation in primary malignant melanoma lesions: association with lack of spontaneous regression. AB - Recently, expression of the immunoproteasome subunits low molecular protein (LMP) 2 or LMP7 was shown to reduce the presentation of certain major histocompatibility complex (MHC) class I-restricted tumour peptide epitopes in renal cell carcinoma and melanoma cells. This may provide the tumour cells with an immune escape mechanism. To test the relevance of this hypothesis, we have taken advantage of the fact that spontaneous regression of human primary melanoma is thought to be the result of a successful peptide-specific cellular immune response in vivo. Immunohistochemical staining with anti-LMP2 and anti-LMP7 xenoantibodies showed a significantly higher expression of these immunoproteasome subunits in primary melanoma lesions exhibiting histological signs of tumour regression than in primary melanoma lesions without regression phenomena. In spontaneously regressing melanoma lesions, LMP2 and LMP7 expression was significantly associated with the presence of tumour-infiltrating lymphocytes. Our results are compatible with the possibility that the expression of the immunoproteasome subunits LMP2 and LMP7 rather than their downregulation in melanoma cells is associated with the presence of a successful anti-melanoma immune response. PMID- 12883364 TI - Epidermal hyperplasia overlying human melanoma correlates with tumour depth and angiogenesis. AB - The aim of this study was to determine whether epidermal hyperplasia overlying cutaneous human melanoma is associated with increased tumour angiogenesis, tumour growth and the potential for metastasis. Forty-two surgical specimens of cutaneous human melanoma of different depths, each containing epidermis present in the tumour-free margin, were analysed by immunohistochemistry for the expression of the pro-angiogenic molecules basic fibroblast growth factor (bFGF), vascular endothelial growth factor (VEGF) and interleukin-8 (IL-8) and the anti angiogenic molecule interferon-beta (IFN-beta). The epidermis overlying intermediate and thick (1.0-10.0 mm), but not thin (0.5-1.0 mm), melanoma specimens was hyperplastic. Although the expression level of bFGF, VEGF and IL-8 in the epidermis directly overlying the tumour was similar to that in the distant epidermis, the expression of IFN-beta was significantly decreased in keratinocytes overlying intermediate and thick, but not thin, melanomas. The microvessel density was also increased in intermediate and thick specimens. Human melanoma cells were injected subcutaneously into nude mice. The resulting tumours were used to determine the association between overlying epidermal hyperplasia and neoplastic angiogenesis. Similar to human autochthonous melanomas, epidermal hyperplasia was found only over lesions produced by metastatic cells. Although there was no change in the expression of the pro-angiogenic molecules, the expression of IFN-beta was significantly decreased in the hyperplastic epidermis. Conditioned medium collected from cultures of the metastatic cell line induced in vitro proliferation of mouse keratinocytes, whereas conditioned medium collected from cultures of the non-metastatic cell line did not. Collectively, the data demonstrate that metastatic melanoma cells induce keratinocyte proliferation, leading to decreased expression of the negative regulator of angiogenesis, IFN beta, and hence to increased angiogenesis. PMID- 12883365 TI - A randomized phase II trial of SRL172 (Mycobacterium vaccae) +/- low-dose interleukin-2 in the treatment of metastatic malignant melanoma. AB - We conducted a randomized phase II trial of SRL172 (Mycobacterium vaccae) +/- low dose interleukin-2 (IL-2) as treatment for stage IV malignant melanoma. The objectives were to establish the safety and efficacy of SRL172 with and without IL-2. All patients had measurable metastatic disease and none received concurrent chemotherapy, radiotherapy, corticosteroids or any other investigational agent. Sixteen patients were randomized into each arm of the trial prior to closure. The trial was halted prematurely when no responses were seen in the first 16 patients receiving SRL172 alone, predicting a response rate of less than 20%. Three partial remissions were seen in the 16 patients receiving SRL172 + IL-2. These patients remained on monthly SRL172 + IL-2, with disease progression at 12, 15 and 23 months. They continued on the trial regimen following surgical management of their disease progression. This trial provides preliminary evidence of a new, non-toxic, immunotherapeutic regimen in the management of malignant melanoma. Further trials are required to establish a definitive response rate and to compare the combination regimen with the regimen of low-dose IL-2 used in this trial. A biological basis for the responses seen in the SRL172 + IL-2 arm also needs to be established. PMID- 12883366 TI - Long-term results of a double perfusion schedule using high dose hyperthermia and melphalan sequentially in extensive melanoma of the lower limb. AB - The aim of this study was to assess the results of an isolated limb perfusion (ILP) schedule with high dose hyperthermia (42-43 degrees C) and melphalan, applied sequentially in patients with advanced melanoma of the limbs. Seventeen patients with extensive recurrent or bulky melanoma of a limb were treated with hyperthermic femoral ILP (42-43 degrees C) without drugs followed by normothermic (37-38 degrees C) ILP with melphalan. Eleven patients (65%) had a complete response. Three patients (27%) had limb recurrences after 5, 6 and 18 months, respectively. The 5 year limb recurrence-free interval for patients with a complete response was 63%. Limb toxicity was mild; pressure-related blistering and transient sensory disturbances occurred after the hyperthermic ILP, and 88% of the patients had a grade II reaction (mild erythema and oedema) after the second ILP. This sequential ILP schedule resulted in a high complete response rate and a low limb-recurrence rate in patients with extensive, recurrent melanoma of the limbs at the cost of only mild toxicity. This regimen could be an alternative to ILP with tumour necrosis factor-alpha and melphalan. PMID- 12883367 TI - On the effect of biochemotherapy in metastatic malignant melanoma: an immunopathological evaluation. AB - Although immunotherapy and biochemotherapy have shown promise, producing a subset of durable responses, for the majority of patients with metastatic melanoma the prognosis is still poor. Therefore there is a great need for predictive tests to identify patients with a high probability of responding. Furthermore, there is also a need for a better understanding of the mechanisms of action during treatment in order to be able to monitor the relevant antitumour reactivity during treatment and to optimize the efficacy of future immunotherapy and biochemotherapy. In the present study histopathological regression criteria were used to study the efficacy of biochemotherapy. Thirty-two patients with metastatic malignant melanoma (18 with regional disease and 14 with systemic disease) were treated with biochemotherapy (cisplatin 30 mg/m2 intravenously on days 1-3, dacarbazine 250 mg/m2 intravenously on days 1-3 and interferon-alpha2b 10 million IU subcutaneously 3 days a week, every 28 days). Pre-treatment fine needle aspirates were obtained from metastases to analyse the number of tumour infiltrating CD4+ lymphocytes. Therapeutic efficacy was evaluated in metastases resected after treatment using histopathological criteria of tumour regression. Comparisons were also made with metastases from 17 untreated patients, all with regional disease. Regressive changes of 25% or more (of the section area) were found in two of the 17 untreated patients with regional disease compared with 13 of the 18 patients with regional disease and 10 of the 14 patients with systemic disease after biochemotherapy. Fifty per cent of the patients with regional disease showed a high degree of regressive changes (75-100% of the section area) after biochemotherapy. These results demonstrate the occurrence of an antitumour reactivity in the majority of patients. Patients with extensive regressive changes in 75-100% of the analysed biopsies were also found to have a longer overall survival (P = 0.019). In patients with regional disease there was a close correlation between a larger number of CD4+ lymphocytes pre-treatment and a higher degree of regressive changes post-treatment (P < 0.05). Thus, immunohistochemical analysis of tumour biopsies shortly after treatment seems to be a good surrogate endpoint. This technique also allows detailed analysis of antitumour reactivity and escape mechanisms. PMID- 12883368 TI - Ocular melanoma is not associated with CDKN2A or MC1R variants--a population based study. AB - Germline variants in the melanocortin 1 receptor gene (MC1R) and the p16 gene (CDKN2A) are associated with an increased risk of cutaneous melanoma. The frequency of these germline variants was examined in a population-based, incident series of 62 ocular melanoma cases and ethnicity-matched population controls. In both cases and controls, 59% of individuals carried at least one MC1R variant and there were no significant differences in the frequency of any of the five most common variants of MC1R. We also found no significant differences between cases and controls in the frequency of any of the four most common variants of CDKN2A, and no melanoma case carried a deleterious germline CDKN2A mutation. Our findings argue against an important predisposing effect of the MC1R and CDKN2A genes for ocular melanoma. PMID- 12883369 TI - Malignant melanoma in South Africans of mixed ancestry: a retrospective analysis. AB - Little is known about the behaviour of melanoma in patients of mixed ancestry. A retrospective analysis of 844 consecutive patients presenting with melanoma over a 12-year period was performed. Forty patients (4.8%) were of mixed ancestry. The data evaluated included patient age, gender, delay in presentation, presenting stage, anatomical distribution, histology, management and outcome. The mean age at presentation was 52.8 years. Twenty-seven patients were female. The mean delay in presentation was 1.54 years. Seventy per cent of melanomas were confined to the extremities, of which one-third were plantar in origin. The most common histological variant, affecting 13 patients (32.5%), was acral lentiginous melanoma; 12.5% of patients presented with in situ (Stage 0) disease, 17.5% with Stage I disease, 22.5% with Stage II disease, 27.5% with Stage III disease and 7.5% with Stage IV disease. Twenty-seven patients (67.5%) remained alive at the end of the study after a median follow-up of 5.58 years, whilst 11 (27.5%) died after a median of 2.42 years. The median survival was 3.92 years. Although the histological type and anatomical distribution reflect the disease pattern of black populations, the overall 5-year survival of 74% is similar to that seen in white populations. An education programme is needed to improve melanoma awareness in mixed race populations. PMID- 12883370 TI - Skin cancer in children and young adults: 28 years' experience from the Northern Region Young Person's Malignant Disease Registry, UK. AB - Little population-based data has been published about skin cancers in children and young adults. In this study, 200 cases of melanoma and non-melanoma skin cancers diagnosed under 25 years of age in the North of England from 1968-1995 were obtained from the Northern Region Young Persons' Malignant Disease Registry. The incidence was 1.2 cases per million per year for children (aged 0-14 years) and was 13 cases per million per year for young adults (aged 15-24 years). Melanoma accounted for 138 cases, of which 16 were in subjects aged < 15 years at diagnosis. The incidence of melanoma increased in females at a rate of 5.6 per million per decade (95% confidence interval [CI] 2.2-8.9, P = 0.002), largely due to an increased incidence of primary lower limb tumours. The incidence for males was unchanged. Survival improved significantly over time for both males and females (P < or = 0.02). Of the 62 patients with non-melanoma skin cancers, 66% were diagnosed with primary non-basal cell carcinoma, 13% with dermatofibrosarcoma protuberans, 10% with squamous cell carcinoma and 11% with other tumours. Two cases were iatrogenic second malignancies following treatment for an earlier primary brain tumour. The incidence of non-melanoma skin cancers was significantly higher during 1982-1995 than during 1968-1981 (rate ratio 1.7, 95% CI 1.0-2.8). There were three deaths from non-melanoma skin cancer, and the overall 5 year survival rate was 98% (95% CI 89-100%). The reason for the increasing incidence of both melanoma and non-melanoma skin cancer in young people is unknown, but it is likely that ultraviolet exposure plays an aetiological role. It is important that families continue to be advised of the need for vigilance with regard to childhood sun exposure. PMID- 12883371 TI - S100 serum level: a tumour marker for metastatic melanoma. PMID- 12883372 TI - Virus therapy of human cancers. PMID- 12883373 TI - Atrioventricular nodal blocking secondary to melanoma infiltration. PMID- 12883374 TI - New avenues in cancer prevention. PMID- 12883375 TI - Occupational exposure to polychlorinated biphenyls and cancer risk. AB - Polychlorinated biphenyls (PCBs) are a class of chemicals characterized by a long term persistence and diffusion in the environment, and by bioaccumulation through the food chain. The highest exposures occurred in occupational cohorts through inhalation or skin absorption in work environments. These cohorts can therefore provide important information on health risk from exposure to PCBs in the general population. To provide comprehensive evidence on cancer risk from PCB exposure, we have thus reviewed the epidemiological studies on workers occupationally exposed to PCBs. Overall, no excess for all cancer mortality was observed in the six studies providing information (573 cancer deaths versus 630.4 expected, corresponding to a standardized mortality ratio (SMR) of 91). Among neoplasms potentially related to PCB exposure, there were 12 deaths from liver cancer compared with 9.5 expected (SMR=126). No excess was found for cancers of the breast (40 observed versus 47.4 expected, SMR=84) and of the lymphatic and haematopoietic system (51 observed versus 53.2 expected, SMR=96). Therefore, studies on occupational exposure to PCBs do not show any excess in all cancer mortality, or in mortality for specific cancer sites of interest. PMID- 12883376 TI - Dose-response relationship between tobacco and lung cancer: new findings. AB - The purpose of this work is to model the relationship between smoking-related variables and the risk of lung cancer by using parametric and non-parametric models. A hospital-based case-control study was conducted to ascertain the influence of smoking on risk of lung cancer. We used parametric logistic regression with a series of categorized independent variables and non-parametric logistic regression models. Such models allow for variables to be treated as continuous, since they avoid arbitrariness in the selection of cut-offs and furnish information on the dose-risk relationship. The results point to the possible existence of a saturation effect for a lifetime tobacco consumption of around 25 000-30 000 packets. Duration of habit and years of abstinence show a linear relationship marked by opposite, though similar, slopes, which would seem to indicate that for every year of smoking, risk rises by an amount (8.00%, 95% confidence interval (CI) 5.94-10.06) equal to the decline in risk for every year of abstinence (6.98%, 95% CI 2.53-11.84). Lastly, a lower age of smoking initiation appears to have an influence, although non-significant, on the appearance of the disease. The risk of lung cancer due to duration of the habit would seem to be proportional to years of abstinence, and there could be a saturation effect with respect to lifetime tobacco consumption. PMID- 12883377 TI - The recent decline in gallbladder cancer mortality in Europe. AB - Mortality from gallbladder cancer has been traditionally high in Eastern Europe, and lower in northern countries. Trends in 18 European countries, including the European Union (EU) and selected Eastern European countries, have been updated using official death certification data abstracted from the WHO database over the period 1980-1999. In the EU, age-standardized rates declined by about 30% between the late 1980s and 1999 to reach 1.8/100 000 for women, and by about 10% to reach 1.4/100 000 for men. In the Czech Republic and Hungary, rates for women were over 6/100 000 until the early 1990s, and declined by about 25% thereafter. For males, gallbladder cancer mortality showed no consistent trend, with rates over 3/100 000. Thus, a high mortality area from gallbladder cancer is still evident for both sexes in Central and Eastern Europe. The trends in mortality from gallbladder cancer are probably influenced by changes in risk factor exposure, such as diet, nutrition or tobacco, but essentially reflect more widespread and earlier adoption of cholecystectomy in the EU, since gallstones are the major risk factor for gallbladder cancer. The data also indicate the scope for further improvement of the management of gallbladder disease in Eastern Europe. PMID- 12883378 TI - Lifestyle, parity, menstrual and reproductive factors and risk of gallbladder cancer. AB - Together with thyroid cancer, cancer of the gallbladder is the only non-sex hormone-related cancer displaying a female preponderance, with incidence being 3 4 times more common among women. We carried out this study to evaluate the role of menstrual, reproductive and lifestyle factors in gallbladder carcinogenesis. A case-control study involving 64 newly diagnosed cases of gallbladder cancer and 101 cases of cholelithiasis was carried out. A detailed menstrual and reproductive history was illustrated beside detailed lifestyle history, in particular consumption of betel nut, tobacco and alcohol and smoking, odds ratio was calculated. Mean age of the patients with cancer was 51+/-1.2 years while it was 40.9+/-1.2 years for gallstones; 69% of cancer patients and 90% of gallstones patients were females. More than half of the cancer patients (53%) and 43% of the gallstone patients were illiterate. A past history of typhoid was present in 22% of cancer patients and 13% of gallstone patients, while 35% of cancer and 25% of gallstone patients were chewers, 18.1 and 9.9% were smokers, and 10% of cancer and 2% of gallstone patients consumed alcohol. Mean age of menarche was 13.4+/ 1.2 years among female patients with cancer while it was 14.0+/-1.4 years for gallstone patients. Higher age at menarche (>13 years, OR 2.48, 95% confidence interval (CI) 1.16-5.3), higher number of childbirths(>3 births, OR 3.92; 95% CI 1.4-10.3), higher number of pregnancies (>3 pregnancies, OR 6.66, 95% CI 1.8 23.4), and higher age at last childbirth (>25 years, OR 2.97, 95% CI 1.04-8.5) were found to have significantly higher risk of developing gallbladder cancer. In conclusion, tobacco chewing and smoking are associated with increased odds of gallbladder cancer. Similarly early menarche, late menopause, multiple pregnancies and childbirth increased the risk of gallbladder cancer. PMID- 12883379 TI - Twenty-year trends of primary liver cancer incidence rates in an urban Chinese population. AB - The objective of this study was to describe trends in the incidence rates of primary liver cancer in a geographically defined Chinese population. Primary liver cancer cases (N=13 685) were diagnosed between 1981 and 2000 and identified by the Tianjin Cancer Registry. Age-adjusted and age-specific incidence rates were examined in both males and females. Poisson regression was employed to assess the incidence rate trends. Crude and age-adjusted incidence rates in the study period were: 27.4/100 000 and 16.4/100 000 in males and 11.5/100 000 and 6.4/100 000 in females, respectively. While the results from Poisson regression analyses suggest statistically significant trends of declining incidence rates of primary liver cancer overall, trends were not consistent across age and sex groups. The decline in incidence was observed, for the most part, in the 40-69 age group, with a greater decrease in males. Our findings provide a new evidence of a downward trend in incidence rates of this disease in China for a period of 20 years. As the observed decline is relatively small and inconsistent across sex and age groups, a continued epidemiological observation on this condition is required. PMID- 12883380 TI - Moderate exercise training slows mammary tumour growth in adolescent rats. AB - Adolescence and young adulthood may be critical windows in establishing risk for breast cancer development in humans. Epidemiological data suggest that exercise during this life stage is associated with decreased breast cancer risk yet few experimental studies to elucidate the mechanism have been performed. The purpose of these studies was to evaluate the effects of moderate exercise training on mammary tumour development in adolescent rats using the 1-methyl 1-nitrosourea (MNU) chemical carcinogen model. Exercise (EX) consisted of moderate-intensity treadmill running 30 min/day, 5 days a week. A total of 274 animals were used: 94 in study 1 and 180 in study 2. Animals were injected with MNU (50 and 25 mg/kg body weight in studies 1 and 2, respectively) at 21 days of age and began training at 28 days of age. Groups of animals (n=10-30 depending on the study and time point) were sacrificed every 2 weeks for 8 weeks to evaluate tumour development. No difference in median tumour-free survival time was observed in the EX versus sham-exercise (SHAM), nor were there any differences in multiplicity at either a high or moderate dose of MNU. Latency to first tumour palpated was increased in both studies by 3-4 days. Consistent across both studies, tumour weights were less and the growth rates of the tumours, defined as tumour weight divided by the number of days elapsed since the tumour was first palpated, were reduced in the EX group. The data suggest that latency is increased and tumour growth is retarded in response to moderate exercise training. PMID- 12883381 TI - Cost-effectiveness of a systematic training programme in breast self-examination. AB - Breast self-examination (BSE) appears to be an effective method for earlier detection of breast cancer. The provision of specialized training programmes in BSE may increase the awareness of breast cancer as well as improve the practice of BSE. This study assessed the cost-effectiveness of a BSE programme that a Danish health authority is considering implementing. A model was developed that related the cost of training (of a specified target group) to the anticipated effects of the training. These anticipated effects included increased knowledge of BSE, more routine use of BSE, fewer cases diagnosed with breast cancer where the tumour is larger than 20 mm and fewer cases where the cancer has spread to lymph nodes. The model was calibrated with data synthesized from the literature, own survey data and local expert guess. In the baseline scenario, the incremental cost per person with better knowledge of BSE was estimated to be 106 euro (2002 price level), the cost per person who begins to undertake BSE to be 180 euro, and the cost per person who correctly applies the technique to be 142 euro. The cost per avoided cancer with spreading to lymph nodes was estimated to be 15 410 euro and the cost of avoiding a cancer tumour larger than 20 mm to be 16 318 euro. Sensitivity analysis showed that these cost-effectiveness ratios varied with different assumptions for the effects of BSE training. PMID- 12883382 TI - Evaluation of an algorithm to identify incident breast cancer cases using DRGs data. AB - Hospital databases have the potential to be inexpensive, timely and nationally representative sources of information about cancer. This study examines the utility of the French hospital database adapted from the Diagnosis Related Group (DRG) classification and named 'Programme de medicalisation des systemes d'information (PMSI)', as an independent source to identify incident cancer cases. From the 19 679 women hospitalized and treated in 1998 in the public hospitals of the Somme area in France, we identified those diagnosed with breast cancer in the PMSI database. These women were matched with women in the cancer registry of the Somme area who had been diagnosed with breast cancer in 1998. An algorithm was used to identify cancer-related diagnoses and procedures reported to PMSI. The sensitivity, specificity and positive predictive value (PPV) of the PMSI database were calculated using the cancer registry as a gold standard. The PMSI database had 85% sensitivity, 99.9% specificity and 97% PPV for women hospitalized with breast cancer as a principal diagnosis. The sensitivity was higher by 9% for hospitalization with breast cancer as a secondary diagnosis but had a lower PPV (78%). In conclusion, the PMSI database seems to offer an interesting potential to assess breast cancer incidence, because of its high sensitivity, in particular when secondary diagnosis was considered, and its very high specificity and PPV. However, these preliminary results need to be confirmed by other studies in France before such databases are used, particularly in areas without cancer registries. PMID- 12883383 TI - Follow-up of radio and telegraph operators with exposure to electromagnetic fields and risk of breast cancer. AB - It is still unclear whether exposure to electromagnetic fields (EMF) is associated with breast cancer. To further investigate the issue, we followed-up a cohort of Norwegian female radio and telegraph operators in the period 1 January 1961 to 31 May 2002, with 99 breast cancer cases. The standardized incidence ratio (SIR) for breast cancer was 1.30 (95% confidence intervals (CI) 1.05-1.58), compared with the total Norwegian female population. In a subsequent nested case control study, exposure to radio frequency (405 kHz-25 MHz) and extremely low frequency (50 Hz) fields due to stay in the radio room during day and night was cumulated by years of employment and workload according to ship type. The exposure was assessed in two age groups (<50, 50+) with regard to risk of breast cancer. The odds ratios in the group with the highest cumulative exposure were 1.78 (95% CI 0.59-5.41) and 2.37 (95% CI 0.88-6.36) in the younger and the older women, respectively. P-value for trend was 0.03 in both age groups. The results of the oestrogen receptor status analysis by exposure to EMF showed an increased risk of oestrogen receptor-positive breast cancer in the younger women, while the older age group had an elevated risk of oestrogen receptor-negative breast cancer. Thus, the present study contributes to the hypothesis of an association between occupational exposure to EMF and increased risk of breast cancer. PMID- 12883384 TI - Interrelationships between plasma testosterone, SHBG, IGF-I, insulin and leptin in prostate cancer cases and controls. AB - Despite strong indirect evidence that androgens stimulate prostate cancer development, data from most analytical studies on this association have been negative. To further investigate this issue, we studied the interrelationships between androgenicity and insulin-like growth factor I (IGF-I), insulin and leptin. Within a prospective cohort study, we measured testosterone, sex hormone binding globulin (SHBG) and IGF-I, IGF-binding protein (IGFBP)-1, IGFBP-3, insulin and leptin, in plasma from 149 cases and 298 controls. Testosterone correlated positively with SHBG, whereas testosterone and SHBG correlated inversely with IGF-I, IGFBP-3, insulin, leptin and body mass index (BMI). Indices of free testosterone showed an inverse linear correlation with leptin (P<0.01), and a strong drop in the 5th quintile of BMI. However, levels of free testosterone showed non-linear relationships over quintiles of insulin and IGF-I, with a significant increase in the second quintile of IGF-I compared with other levels. The absence of an association between plasma levels of androgens and prostate cancer risk in analytical studies, despite the strong indirect evidence of their tumour-stimulating effects, may reflect the complex and mostly inverse associations of androgenicity to IGF-I, insulin and leptin which are hormones that have also been implicated as risk factors for prostate cancer. PMID- 12883385 TI - Polymorphisms in glutathione-S-transferase genes (GST-M1, GST-T1 and GST-P1) and susceptibility to prostate cancer among male smokers of the ATBC cancer prevention study. AB - Glutathione-S-transferase (GST) genes encode a family of detoxification enzymes that offer protection against endogenous and exogenous sources of reactive oxygen species (ROS). Germline variations in GST genes may alter the catalytic efficiency of GST isoenzymes leading to a potential increase in susceptibility to the genotoxic effects of ROS and electrophilic substances. A nested case-control study design was used to examine the association between the polymorphic GST genes and prostate cancer risk among Finnish male smokers of the ATBC Cancer Prevention Study. A case-case analysis was used to determine the association between these genetic polymorphisms and prostate cancer progression. Germline DNA was obtained from 206 prostate cancer cases and 194 controls frequency matched on age, intervention group and study clinic. Cases and controls were genotyped for three GST genes using MALDI-TOF mass spectrometry or multiplex polymerase chain reaction (PCR). Relative to the wild-type genotype, we observed a 36% reduction in prostate cancer risk associated with the GST-M1-null genotype (odds ratio (OR) 0.64, 95% confidence interval (CI) 0.43, 0.95). Unlike GST-M1, GST-T1-null (OR 0.74, 95% CI 0.42, 1.33) and GST-P1*B (OR 1.10, 95% CI 0.72, 1.69) were not strongly associated with prostate cancer risk. We did not observe any significant associations between the selected polymorphic GST genes and tumour grade or stage. In conclusion, we did not observe a direct association between polymorphic GST-T1 or GST-P1 and prostate cancer risk. Our observation of a relatively strong inverse association between the GST-M1-null genotype and prostate cancer risk needs to be confirmed in larger association studies. PMID- 12883386 TI - Plasma concentrations of coenzyme Q10 and tocopherols in cervical intraepithelial neoplasia and cervical cancer. AB - Cervical intraepithelial neoplasia (CIN) may, at times, unpredictably progress to invasive carcinoma of the cervix. Epidemiological nutritional studies suggest that higher dietary consumption and circulating levels of certain micronutrients may be protective against cervical cancer. However, a preventive role of dietary antioxidants in CIN is not well established. The purpose of this cross-sectional study was to investigate the comparative plasma concentrations of three potent antioxidants, coenzyme Q(10,) alpha-tocopherol and gamma-tocopherol, in women with normal Pap smears and patients with a biopsy-confirmed histopathological lesion diagnosed as CIN or cervical cancer. Plasma concentrations of coenzyme Q(10,) alpha-tocopherol and gamma-tocopherol were measured by high-pressure liquid chromatography in both normal women without any history of abnormal Pap smears (n=48), and patients with histopathologically confirmed diagnoses of: (a) CIN I, n=98; (b) CIN II, n=49; (c) CIN III, n=10; and (d) cervical cancer, n=25. The mean plasma levels of coenzyme Q(10), alpha-tocopherol and gamma-tocopherol were significantly lower (P<0.001,<0.001, and<0.001, respectively by Kruskal Wallis test) in patients with various grades of CIN and cervical cancer compared with controls. After controlling for age and smoking, an inverse association between histological grades of epithelial lesions and both plasma coenzyme Q(10) and alpha-tocopherol concentrations was observed. The low plasma concentrations of coenzyme Q(10) may be due to deficient dietary intake or a decrease in endogenous coenzyme Q(10) biosynthesis that may reflect increased utilization as a result of free radical reactive oxygen species induced oxidative stress. Further molecular studies on the mechanistic role of antioxidants in women with precancer cervical lesions are needed. PMID- 12883387 TI - Time trends in cigarette smoking in two German cohorts--results from EPIC Germany. AB - Smoking prevention is less advanced in Germany compared with other European and North American countries, and fewer data exist, especially on the consumption habits at the individual level over time. EPIC Germany, which is part of a European multicentre study on diet and cancer, collected data on individual smoking behaviour and allows for consideration of the changing consumption pattern for both centres and different age groups. Within EPIC 25 546 and 27 548 participants, respectively, were recruited in Heidelberg and Potsdam. Data on smoking habits were collected by means of a computer-guided interview during the baseline examination between 1994 and 1998. For each birth cohort smoking prevalence and mean number of cigarettes smoked per day at different ages were calculated. Additionally, the prevalence of non-filter cigarette smoking was computed. Smoking prevalence in the 1990s was still higher among men (Heidelberg 16.3-32.3%; Potsdam 18.2-29.3%) than among women (Heidelberg 12.8-32.0%; Potsdam 10.4-27.8%). However, the percentage of women smokers was still increasing. Filter cigarettes comprised a growing percentage of the cigarettes smoked, but especially among men differences between both German cohorts can still be seen: depending on age, 10.0-12.7% of men in the Heidelberg cohort smoked non-filter cigarettes, but only 1.1-2.3% in the Potsdam cohort. The quantity smoked was higher in the Heidelberg than in the Potsdam cohort with respect to the mean number of cigarettes smoked per day as well as the pack-years of smoking. In conclusion, smoking habits in the Potsdam and the Heidelberg cohorts did not strongly differ by smoking prevalence. However, they did differ according to the quantity and quality of smoking. These differences, as well as the changes over the last 40 years may contribute to a changing pattern of diseases in different groups of the German population. PMID- 12883388 TI - Price and consumption of tobacco in Italy over the last three decades. AB - Changes in cigarette price have had an appreciable impact on smoking consumption in several countries. We analysed the price elasticity of demand for cigarettes in Italy over the period 1970-2001. A 1% increase in price of cigarettes led to a 0.30% decline in smoking prevalence and to a 0.43% decline in cigarette consumption. The present analyses confirm the existence of an inverse association between price and prevalence/consumption of cigarettes in Italy, and indicate that economic aspects may have important public health implications in tobacco control. PMID- 12883389 TI - The relationship of physical activity with dietary cancer-protective nutrients and cancer-related biological and lifestyle factors. AB - The aim of the present study was to analyse the relationships among different intensities of leisure-time physical activity (PA) and cancer-protective dietary components, cancer-related biological (general and central obesity) and lifestyle factors (heavy smoking and excessive alcohol consumption). The present population based cross-sectional study (765 men and 812 women from the general population of Gerona, Spain) included measurement of anthropometrical data, dietary intake, and alcohol and cigarette consumption and furthermore detailed evaluation of total leisure-time physical activity, including measurement of different intensities. Logistic regression analysis adjusted for several confounders was used to analyse the relationships among variables. A composite variable composed of dietary fibre, vitamin C, alpha-tocopherol, beta-carotene and folate was directly associated with higher levels of leisure-time PA in both genders, after further adjusting for energy intake [odds ratio (OR) of low versus second tertile of total leisure-time PA was 1.98 (95% confidence interval (CI) 1.22-3.23) for men and OR of low versus top tertile of total leisure-time PA was 1.75 (95% CI 1.07 2.84) for women]. Low to moderate- but not high-intensity leisure-time PA was directly related to higher intakes of these nutrients. Heavy smoking was inversely associated with higher levels of total leisure-time PA in men. With the exception of central obesity in women, there was no significant relationship between the cancer-related biological and lifestyle factors, and low to moderate intensity leisure-time PA in both genders. In contrast, central obesity (OR 0.31, 95% CI 0.16-0.63) and heavy smoking (OR 0.38, 95% CI 0.16-0.96) were significantly related to high-intensity but not to total or low to moderate intensity leisure-time PA in men and women, respectively. The observed relationships between leisure-time PA and several of the studied cancer associated variables might partially account for the protective effect of physical activity against cancer observed in the literature. However, different intensities of leisure-time PA were distinctly associated with the mentioned cancer-related dietary, biological and lifestyle factors. Most importantly, although some of these variables were not associated with total leisure-time PA a significant relationship was observed for intensities of leisure-time PA. This should be considered when analysing the relationship between leisure-time PA and cancer. PMID- 12883390 TI - Trends in testicular cancer incidence in Vaud, Switzerland. AB - Upward trends in testicular cancer incidence have been reported in Europe and North America, particularly for seminomas. We considered incidence data between 1974 and 1999 from the Swiss cancer registry of Vaud, i.e. one of the highest incidence areas on a worldwide scale, including a total of 731 cases. Testicular cancer incidence was around 8.5/100 000 between the mid-1970s and the late 1980s, and increased to around 10/100 000 in the 1990s. Corresponding figures at age 15 44 were around 16/100 000 between the mid-1970s and the late 1980s, and about 19/100 000 thereafter. No evidence of persisting upward trends was evident over the last few years. The rise in testicular cancer incidence in the 1990s was apparently restricted to seminomas, whose rates increased from about 4 to 5.7/100 000 at all ages, and from 7 to over 11/100 000 at age 15-44. No consistent pattern of trends was observed for malignant teratomas and for other and unspecified histotypes. Testicular cancer in Vaud has shown no tendency to further rising over the last decade, thus re-opening the issue of a probable asymptote of testicular cancer incidence in this population. PMID- 12883391 TI - Are scientists normal? PMID- 12883393 TI - Is routine endotracheal intubation as safe as we think or wish? PMID- 12883394 TI - Apples and oranges: the fruits of labor in anesthesia care. PMID- 12883395 TI - Is regional anesthesia simply an exercise in applied sonoanatomy?: aiming at higher frequencies of ultrasonographic imaging. PMID- 12883396 TI - Laryngeal resistance before and after minor surgery: endotracheal tube versus Laryngeal Mask Airway. AB - BACKGROUND: The placement of an endotracheal tube (ETT) may promote laryngeal swelling, which is an important cause of upper airway obstruction after extubation. The authors hypothesized that laryngeal swelling after ETT placement increases laryngeal resistance and tested that hypothesis by comparing postoperative laryngeal patency between patients with ETT placement and those with a Laryngeal Mask Airway trade mark (LMA). METHODS: Fourteen adult patients who underwent elective minor surgeries were randomly allocated to two groups whose airway would be managed through ETTs (the ETT group) or LMAs (the LMA group) during the surgery. While maintaining at sevoflurane 1 minimum alveolar concentration, the authors measured laryngeal resistance before and after surgery, during both spontaneous breathing and mechanical ventilation under complete paralysis. In addition, they endoscopically measured the vocal cord angle under complete paralysis. RESULTS: In association with marked swelling of the vocal cords, the vocal cord angle significantly decreased after surgery in the ETT group, whereas the angle did not change in the LMA group. Laryngeal resistance during mechanical ventilation significantly increased only in the ETT group. Laryngeal resistance during spontaneous breathing significantly increased after surgeries in both groups. CONCLUSIONS: Postoperative laryngeal resistance increases at least in part because of laryngeal swelling in patients with ETT placement, whereas alteration of laryngeal neural control mechanisms has been also indicated. The use of the LMA trade mark has an advantage over ETT placement in order to avoid postoperative laryngeal swelling. PMID- 12883397 TI - Predictors of survival following cardiac arrest in patients undergoing noncardiac surgery: a study of 518,294 patients at a tertiary referral center. AB - BACKGROUND: The authors determined the incidence of cardiac arrest and predictors of survival following perioperative cardiac arrest in a large population of patients at a tertiary referral center. METHODS: Medical records of patients who experienced cardiac arrest in the perioperative period surrounding noncardiac surgery between January 1, 1990, and December 31, 2000, were reviewed. Logistic regression identified characteristics associated with immediate (>or= 1 h) and hospital survival, with P /= 13 ng/ml). In-hospital mortality and nonfatal cardiac events (delayed extubation > 24 h; postoperative requirement of inotropic agent; ventricular and supraventricular arrhythmia; postoperative myocardial infarction) were recorded. Survivors were then followed up over a 2-yr period. Data are median and odds ratio (95% confidence interval). RESULTS: Of all patients, 174 (86%) had a low cTnI (4.1 ng/ml; range, 1.1-12.6) and 28 (14%) had a high cTnI (23.8 ng/ml; range, 13.4-174.6). In-hospital mortality was not significantly different (4 vs. 2%), whereas long-term mortality (18 vs. 3%, P = 0.006) and mortality from cardiac cause (18 vs. 1%, P < 0.001) was greater in patients with a high cTnI. A high cTnI was a significant factor predicting death (odds ratio, 7.3 [2.0-27.1]) or death from cardiac causes (odds ratio, 37.4 [4.2-334.4]). Nonfatal cardiac events were also more frequent in the hospital (64 vs. 41%, P = 0.02) and within the 2-yr follow-up period (39% vs. 16%, P = 0.03) in patients with high cTnI. CONCLUSION: A high postoperative peak of cTnI is associated with increased risk of death, death from cardiac causes, and nonfatal cardiac events within 2 yr after coronary artery bypass grafting. PMID- 12883399 TI - Pharmacokinetics of midazolam in neonates undergoing extracorporeal membrane oxygenation. AB - BACKGROUND: Although the pharmacokinetics of midazolam in critically ill children has been described, there are no such reports in extracorporeal membrane oxygenation. METHODS: The pharmacokinetics of midazolam and 1-hydroxy midazolam after continuous infusion (50-250 microg. kg(-1). h(-1)) were determined in 20 neonates undergoing extracorporeal membrane oxygenation. Patients were randomized into two groups: group 1 (n = 10) received midazolam extracorporeally (into the circuit), and group 2 received drug via central or peripheral access. Blood samples for determination of plasma concentrations were taken at baseline, 2, 4, 6, 12, 18, and 24 h, then every 12 h. Population pharmacokinetic analysis and model building was conducted using WinNonMix (Pharsight Corporation, Mountain View, CA). The 1-hydroxy midazolam/midazolam metabolic ratio was determined as a surrogate marker of cytochrome P450 3A activity. RESULTS: The parameter estimates (n = 19) were based on a one-compartment model with time-dependent change in volume of distribution. Volume (mean +/- standard error) expanded monoexponentially from the onset of extracorporeal membrane oxygenation to a maximum value, 0.8 l +/- 0.5 and 4.1 +/- 0.5 l/kg, respectively. Consequently, plasma half-life was substantially prolonged (median [range]) from onset to steady-state: 6.8 (2.2-39.8) and 33.3 (7.4-178) h, respectively. Total body clearance was determined as (mean +/- standard error) 1.4 +/- 0.15 ml. kg-1. min 1. The median metabolic ratio was 0.17 (0.03-0.9). No significant differences were observed between the two groups with respect to parameter estimates. Simulations of plasma concentration profiles revealed excess levels at conventional doses. CONCLUSIONS: These results reveal significantly increased volume of distribution and plasma half-life in neonates receiving extracorporeal membrane oxygenation. Altered kinetics may reflect sequestration of midazolam by components of the extracorporeal membrane oxygenation circuit. PMID- 12883400 TI - Performance of noninvasive partial CO2 rebreathing cardiac output and continuous thermodilution cardiac output in patients undergoing aortic reconstruction surgery. AB - BACKGROUND: In the partial CO(2) rebreathing method, monitored changes in CO(2) elimination and end-tidal CO(2) in response to a brief rebreathing period are used to estimate cardiac output. However, dynamic changes in CO(2) production during ischemia and reperfusion may affect the accuracy of these estimates. This study was designed to compare measurements of cardiac output as produced by the partial CO(2) rebreathing (NICO), bolus (BCO), and continuous thermodilution (CCO) methods of monitoring cardiac output. METHODS: Cardiac output was continuously monitored using both NICO and CCO in 28 patients undergoing aortic reconstruction. BCO measurements were taken at the following intervals when hemodynamic stability was achieved: (1) after anesthetic induction; (2) during aortic cross-clamp; (3) at reperfusion of the iliac artery; and, (4) during peritoneal closure. RESULTS: The bias and precision (1 SD) derived from all the measurements between NICO and BCO was -0.58 +/- 0.9 l/min, whereas for CCO and BCO it was 0.38 +/- 1.17 l/min. The bias between NICO and BCO was small after anesthetic induction and during cross-clamp, but increased following reperfusion. The bias between CCO and BCO was relatively small until reperfusion but increased significantly at peritoneal closure. CONCLUSIONS: Results indicate that in aortic reconstruction surgery the performance of NICO monitoring is comparable with that of CCO; however, the direction of bias in these continuous measurement devices is the opposite. PMID- 12883401 TI - Protocol for the sequence analysis of ryanodine receptor subtype 1 gene transcripts from human leukocytes. AB - BACKGROUND: The search for novel mutations in the ryanodine receptor subtype 1 (RYR1) gene causing malignant hyperthermia and central core disease is hampered by the fact that the gene contains 106 exons. Searching for novel mutations in complementary DNA (cDNA) requires an invasive muscle biopsy. Accordingly, an alternate source of RYR1 cDNA was sought for sequence analysis. METHODS: Leukocytes were isolated from human blood and used for extraction of RNA and reverse transcription of messenger RNA into cDNA. A detailed protocol was developed in which overlapping fragments of RYR1 cDNA were amplified by polymerase chain reaction in a series of steps and used for double-strand sequencing. RESULTS: The sequences of full-length leukocyte RYR1 cDNA obtained from four human blood samples were shown to be identical to the sequence of a human muscle RYR1 cDNA. The incidence of aberrant splicing was more pronounced in the blood-derived cDNAs, but this could be minimized by adequate sample preparation. Protocols to sequence alternatively spliced products were also developed. Several silent nucleotide polymorphisms were detected, and minor revisions were made to the RYR1 sequence. CONCLUSIONS: Because there are no differences in RYR1 transcript structure between muscle and leukocytes, aside from those that may be ascribed to RNA splicing aberrations during processing, leukocytes seem to be an adequate substitute tissue for screening the RYR1 gene for previously undiscovered mutations in families with malignant hyperthermia or central core disease. PMID- 12883402 TI - Detection of a novel ryanodine receptor subtype 1 mutation (R328W) in a malignant hyperthermia family by sequencing of a leukocyte transcript. AB - BACKGROUND: To determine whether malignant hyperthermia (MH) susceptibility in a Canadian pedigree is associated with a mutation in the ryanodine receptor subtype 1 (RYR1) gene, the complete RYR1 transcript obtained from the leukocytes of one MH-susceptible family member was sequenced, using a newly developed protocol. METHODS: RNA was extracted from leukocytes and converted into complementary DNA. Overlapping fragments of RYR1 complementary DNA were amplified by the polymerase chain reaction and used for double-strand sequencing to find a single mutation likely to be causal of MH susceptibility. Inheritance of the mutation in the family was studied by restriction endonuclease analysis and/or sequencing of genomic DNA and compared to available caffeine halothane contracture test data. The mutation was introduced into rabbit RYR1 complementary DNA, the complementary DNA was expressed in human embryonic kidney line 293 cells, and Ca2+ release by the mutant Ca2+ release channel was measured following the addition of caffeine and halothane. RESULTS: A novel arginine 328 to tryptophan mutation in RYR1 was detected by direct sequencing of the RYR1 transcript from leukocytes of one MH susceptible individual. A causal role for this mutation in MH is indicated by cosegregation of the mutation with the MH-susceptible phenotype within the family and by the demonstration that the mutant channel has increased sensitivity to both caffeine and halothane. CONCLUSIONS: The feasibility of using complete RYR1 transcripts from leukocytes for sequence analysis offers an efficient and noninvasive method for scanning RYR1 for novel mutations. PMID- 12883403 TI - Pharmacokinetics and clinical pharmacodynamics of the new propofol prodrug GPI 15715 in volunteers. AB - BACKGROUND: GPI 15715 (AQUAVAN injection) is a new water-soluble prodrug which is hydrolyzed to release propofol. The objectives of this first study in humans were to investigate the safety, tolerability, pharmacokinetics, and clinical pharmacodynamics of GPI 15715. METHODS: Three groups of three healthy male volunteers (aged 19-35 y, 67-102 kg) received 290, 580, and 1,160 mg GPI 15715 as a constant rate infusion over 10 min. The plasma concentrations of GPI 15715 and propofol were measured from arterial and venous blood samples up to 24 h. Pharmacokinetics were analyzed with compartment models. Pharmacodynamics were assessed by clinical signs. RESULTS: GPI 15715 was well tolerated without pain on injection. Two subjects reported a transient unpleasant sensation of burning or tingling at start of infusion. Loss of consciousness was achieved in none with 290 mg and in one subject with 580 mg. After 1,160 mg, all subjects experienced loss of consciousness at propofol concentrations of 2.1 +/- 0.6 microg/ml. A two compartment model for GPI 15715 (central volume of distribution, 0.07 l/kg; clearance, 7 ml. kg-1 min-1; terminal half-life, 46 min) and a three-compartment model for propofol (half-lives: 2.2, 20, 477 min) best described the data. The maximum decrease of blood pressure was 25%; the heart rate increased by approximately 35%. There were no significant laboratory abnormalities. CONCLUSIONS: Compared with propofol lipid emulsion, the potency seemed to be higher with respect to plasma concentration but was apparently less with respect to dose. Pharmacokinetic simulations showed a longer time to peak propofol concentration after a bolus dose and a longer context-sensitive half-time. PMID- 12883404 TI - Effects of propofol, desflurane, and sevoflurane on recovery of myocardial function after coronary surgery in elderly high-risk patients. AB - BACKGROUND: The present study investigated the effects of propofol, desflurane, and sevoflurane on recovery of myocardial function in high-risk coronary surgery patients. High-risk patients were defined as those older than 70 yr with three vessel disease and an ejection fraction less than 50% with impaired length dependent regulation of myocardial function. METHODS: Coronary surgery patients (n = 45) were randomly assigned to receive either target-controlled infusion of propofol or inhalational anesthesia with desflurane or sevoflurane. Cardiac function was assessed perioperatively and during 24 h postoperatively using a Swan-Ganz catheter. Perioperatively, a high-fidelity pressure catheter was positioned in the left and right atrium and ventricle. Response to increased cardiac load, obtained by leg elevation, was assessed before and after cardiopulmonary bypass (CPB). Effects on contraction were evaluated by analysis of changes in dP/dt(max). Effects on relaxation were assessed by analysis of the load-dependence of myocardial relaxation. Postoperative levels of cardiac troponin I were followed for 36 h. RESULTS: After CPB, cardiac index and dP/dt(max) were significantly lower in patients under propofol anesthesia. Post CPB, leg elevation resulted in a significantly greater decrease in dP/dt(max) in the propofol group, whereas the responses in the desflurane and sevoflurane groups were comparable with the responses before CPB. After CPB, load dependence of left ventricular pressure drop was significantly higher in the propofol group than in the desflurane and sevoflurane group. Troponin I levels were significantly higher in the propofol group. CONCLUSIONS: Sevoflurane and desflurane but not propofol preserved left ventricular function after CPB in high risk coronary surgery patients with less evidence of myocardial damage postoperatively. PMID- 12883405 TI - Using the time of maximum effect site concentration to combine pharmacokinetics and pharmacodynamics. AB - BACKGROUND: To simulate the time course of drug effect, it is sometimes necessary to combine the pharmacodynamic parameters from an integrated pharmacodynamic pharmacodynamic study (e.g., volumes, clearances, k(e0) [the effect site equilibration rate constant], C(50) [the steady state plasma concentration associated with 50% maximum effect], and the Hill coefficient) with pharmacokinetic parameters from a different study (e.g., a study examining a different age group or sampling over longer periods of time). Pharmacokinetic pharmacodynamic parameters form an interlocked vector that describes the relationship between input (dose) and output (effect). Unintended consequences may result if individual elements of this vector (e.g., k(e0)) are combined with pharmacokinetic parameters from a different study. The authors propose an alternative methodology to rationally combine the results of separate pharmacokinetic and pharmacodynamic studies, based on t(peak), the time of peak effect after bolus injection. METHODS: The naive approach to combining separate pharmacokinetic and pharmacodynamic studies is to simply take the k(e0) from the pharmacodynamic study and apply it naively to the pharmacokinetic study of interest. In the t(peak) approach, k(e0) is recalculated using the pharmacokinetics of interest to yield the correct time of peak effect. The authors proposed that the t(peak) method would yield better predictions of the time course of drug effect than the naive approach. They tested this hypothesis in three simulations: thiopental, remifentanil, and propofol. RESULTS: In each set of simulations, the t(peak) method better approximated the postulated "true" time course of drug effect than the naive method. CONCLUSIONS: T(peak) is a useful pharmacodynamic parameter and can be used to link separate pharmacokinetic and pharmacodynamic studies. This addresses a common difficulty in clinical pharmacology simulation and control problems, where there is usually a wide choice of pharmacokinetic models but only one or two published pharmacokinetic pharmacodynamic models. The results will be immediately applicable to target controlled anesthetic infusion systems, where linkage of separate pharmacokinetic and pharmacodynamic parameters into a single model is inherent in several target controlled infusion designs. PMID- 12883406 TI - Propofol and sufentanil titration with the bispectral index to provide anesthesia for coronary artery surgery. AB - BACKGROUND: To provide anesthesia for cardiac surgery, hypnotics and opioids are frequently titrated on variables such as mean arterial pressure and heart rate. In this study conducted in patients scheduled to undergo coronary artery bypass grafting, propofol and sufentanil, both administered by computer-controlled infusion, were titrated on the Bispectral Index (BIS) values using a predefined algorithm. METHODS: After written informed consent, 110 patients, 95 men and 15 women aged 61 (9) yr [mean (SD)], were randomly allocated to receive predicted sufentanil effect site concentrations (Ce) of 0.5, 0.75, 1, 1.25, and 1.5 ng/ml, decreased by a third after sternotomy (groups 1-5). Target induction propofol concentration was 1.5 microg/ml and subsequently adjusted on BIS values. The following parameters were recorded: BIS values, predicted propofol Ce, the number of changes of propofol target, mean arterial pressure, heart rate, the number of bolus injection and doses of vasoconstrictor and vasodilator drugs, time to tracheal extubation, postoperative awareness and satisfaction scores, and cumulative morphine doses for the first postoperative day. RESULTS: One patient randomized to group 1 required 0.75 ng/ml sufentanil Ce instead of 0.5 ng/ml for increased BIS values on tracheal intubation. BIS values were similar in the five groups. The predicted propofol Ce values were different (P < 0.05; analysis of variance) among the five groups: 1.59 (0.47) to 1.23 (0.25) microg/ml in group 1 and group 4, respectively. Significantly fewer changes of propofol target were required in group 4 as compared to group 1. There were no differences among the five groups for mean arterial pressure, heart rate, time to tracheal extubation, awareness, satisfaction scores, and morphine requirements. CONCLUSION: These results suggest the BIS, as part of an algorithm that uses both the absolute BIS value and its increase following tracheal intubation, can be used to effectively titrate both propofol and sufentanil. A predicted sufentanil Ce of 1.25 ng/ml before and 0.8 ng/ml after sternotomy was associated with the lowest predicted propofol Ce and fewer changes of propofol target. Lower sufentanil concentrations required higher propofol concentrations and more frequent changes of the target propofol concentration and were associated with similar hemodynamic tolerance. PMID- 12883407 TI - Propofol reduces perioperative remifentanil requirements in a synergistic manner: response surface modeling of perioperative remifentanil-propofol interactions. AB - BACKGROUND: Remifentanil is often combined with propofol for induction and maintenance of total intravenous anesthesia. The authors studied the effect of propofol on remifentanil requirements for suppression of responses to clinically relevant stimuli and evaluated this in relation to previously published data on propofol and alfentanil. METHODS: With ethics committee approval and informed consent, 30 unpremedicated female patients with American Society of Anesthesiologists physical status class I or II, aged 18-65 yr, scheduled to undergo lower abdominal surgery, were randomly assigned to receive a target controlled infusion of propofol with constant target concentrations of 2, 4, or 6 microg/ml. The target concentration of remifentanil was changed in response to signs of inadequate anesthesia. Arterial blood samples for the determination of remifentanil and propofol concentrations were collected after blood-effect site equilibration. The presence or absence of responses to various perioperative stimuli were related to the propofol and remifentanil concentrations by response surface modeling or logistic regression, followed by regression analysis. Both additive and nonadditive interaction models were explored. RESULTS: With blood propofol concentrations increasing from 2 to 7.3 microg/ml, the C(50) of remifentanil decreased from 3.8 ng/ml to 0 ng/ml for laryngoscopy, from 4.4 ng/ml to 1.2 ng/ml for intubation, and from 6.3 ng/ml to 0.4 ng/ml for intraabdominal surgery. With blood remifentanil concentrations increasing from 0 to 7 ng/ml, the C(50) of propofol for the return to consciousness decreased from 3.5 microg/ml to 0.6 microg/ml. CONCLUSIONS: Propofol reduces remifentanil requirements for suppression of responses to laryngoscopy, intubation, and intraabdominal surgical stimulation in a synergistic manner. In addition, remifentanil decreases propofol concentrations associated with the return of consciousness in a synergistic manner. PMID- 12883408 TI - Thiopental inhibits tumor necrosis factor alpha-induced activation of nuclear factor kappaB through suppression of kappaB kinase activity. AB - BACKGROUND: Thiopental is frequently used for the treatment of intracranial hypertension after severe head injury and is associated with immunosuppressive effects. The authors have recently reported that thiopental inhibits activation of nuclear factor (NF) kappaB, a transcription factor implicated in the expression of many inflammatory genes. Thus, it was the aim of the current study to examine the molecular mechanism of this inhibitory effect. METHODS: The authors tested gamma-aminobutyric acid (GABA), the GABA(A) antagonist bicuculline, and the GABA(B) antagonist dichlorophenyl-methyl-amino-propyl diethoxymethyl-phosphinic acid (CGP 52432) in combination with thiopental for their influence on the activation of NF-kappaB. In addition, they investigated the direct effect of thiopental on activated NF-kappaB DNA binding activity. These experiments were conducted in Jurkat T lymphocytes using electrophoretic mobility shift assays. The presence of the phosphorylated and dephosphorylated NF kappaB inhibitor IkappaBalpha (Western blotting) and IkappaB kinase activity were studied in Jurkat T cells and human CD3+ T lymphocytes. In addition, the authors tested the effect of the structural barbiturate analog pairs thiopental pentobarbital and thiamylal-secobarbital and of thiopental in combination with the thio-group containing chemical dithiothreitol on the activation of NF-kappaB. RESULTS: GABA did not inhibit NF-kappaB activation, and the GABA(A) and GABA(B) antagonists bicuculline and CGP did not diminish the thiopental-mediated inhibitory effect on NF-kappaB activation. Thiopental did not inhibit activated NF-kappaB directly in a cell-free system. The phosphorylation of IkappaBalpha was prevented after incubation with 1,000 microg/ml thiopental. The same concentration of thiopental also inhibited IkappaB kinase activity in tumor necrosis factor-stimulated Jurkat T cells and human CD3+ T lymphocytes (60% suppression, P < 0.05 vs. tumor necrosis factor alpha alone). Thiobarbiturates (4 x 10(-3) m) inhibited NF-kappaB activity, whereas equimolar concentrations of the structural oxyanalogs did not. Preincubation of thiopental with dithiothreitol diminished the inhibitory effect. CONCLUSION: Thiopental-mediated inhibition of NF-kappaB activation is due to the suppression of IkappaB kinase activity and depends at least in part on the thio-group of the barbiturate molecule. PMID- 12883409 TI - Neuroprotective effects of propofol in a model of ischemic cortical cell cultures: role of glutamate and its transporters. AB - BACKGROUND: During cerebral ischemia, excess of glutamate release and dysfunction of its high affinity transport induce an accumulation of extracellular glutamate, which plays an important role in neuronal death. The authors studied the relationship among propofol neuroprotection, glutamate extracellular concentrations, and glutamate transporter activity in a model of ischemic cortical cell cultures. METHODS: Thirteen-day-old primary cortical neuronal-glial cultures were exposed to a 90-min combined oxygen-glucose deprivation (OGD) in an anaerobic chamber, followed by reoxygenation. Propofol was added only during the OGD period, and its effect was compared to that of the N-methyl-d-aspartate receptor antagonist dizocilpine (MK-801). Twenty-four hours after the injury, cell death was quantified by lactate dehydrogenase release and cell viability by reduction of 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT). Extracellular concentrations of glutamate in culture supernatants and glutamate uptake were performed at the end of OGD period by high-performance liquid chromatography and incorporation of l-[3H]glutamate into cells, respectively. RESULTS: At clinically relevant concentrations (0.05-10 microm), propofol offered protection equivalent to that of MK-801. It significantly reduced lactate dehydrogenase release and increased the reduction of MTT. At the end of the ischemic injury, propofol was able to reverse the OGD-induced increase in glutamate extracellular concentrations and decrease of glutamate uptake. The inhibition of the glial GLT1 transporter by 3-methyl-glutamate did not further modify the effect of propofol on glutamate uptake, suggesting that GLT1 was not the major target of propofol. CONCLUSION: Propofol showed a neuroprotective effect in this in vitro model of OGD, which was apparently mediated by a GLT1 independent restoration of the glutamate uptake impaired during the injury. PMID- 12883410 TI - Spontaneous breathing improves lung aeration in oleic acid-induced lung injury. AB - BACKGROUND: Experimental and clinical studies have shown reduction in intrapulmonary shunt with improved oxygenation by spontaneous breathing with airway pressure release ventilation (APRV) in acute lung injury. The mechanisms of these findings are not clear. The authors hypothesized that spontaneous breathing results in better aeration of lung tissue and that improvement in oxygenation can be explained by these changes. This hypothesis was studied in a porcine model of oleic acid-induced lung injury. METHODS: Two hours after induction of lung injury, 24 pigs were randomly assigned to APRV with or without spontaneous breathing at a positive end-expiratory pressure of 5 cm H(2)O. Hemodynamics, spirometry, and end-expiratory lung volume by nitrogen washout were measured at baseline, after 2 h of lung injury, and after 2 and 4 h of mechanical ventilation in the specific mode. Finally, spiral computed tomography of the chest was performed at end-expiratory lung volume in 22 pigs. RESULTS: Arterial carbon dioxide tension and mean and end-inspiratory airway pressures were comparable between settings. Four hours of APRV with spontaneous breathing resulted in improved oxygenation compared with APRV without spontaneous breathing (arterial oxygen tension, 144 +/- 65 vs. 91 +/- 50 mmHg, P < 0.01 for interaction time x mode), higher end-expiratory lung volume (786 +/- 320 vs. 384 +/- 148 ml, P < 0.001), and better aeration. End-expiratory lung volume and venous admixture were both correlated with the amount of lung reaeration (r(2) = 0.62 and r(2) = 0.61, respectively). CONCLUSIONS: The results support the hypothesis that spontaneous breathing during APRV improves oxygenation mainly by recruitment of nonaerated lung and improved aeration of the lungs. PMID- 12883412 TI - G-protein activation decreases isoflurane inhibition of N-type Ba2+ currents. AB - BACKGROUND: G-protein activation mediates inhibition of N-type Ca2+ currents. Volatile anesthetics affect G-protein pathways at various levels, and activation of G-proteins has been shown to increase the volatile anesthetic potency for inhibiting the electrical-induced contraction in ileum. The authors investigated whether isoflurane inhibition of N-type Ba2+ currents was mediated by G-protein activation. METHODS: N-type Ba2+ currents were measured in the human neuronal SH SY5Y cell line by using the whole cell voltage-clamp method. RESULTS: Isoflurane was found to have two effects on N-type Ba2+ currents. First, isoflurane reduced the magnitude of N-type Ba2+ currents to a similar extent (IC50 approximately 0.28 mm) in the absence and presence of GDPbetaS (a nonhydrolyzable GDP analog). Interestingly, GTPgammaS (a nonhydrolyzable GTP analog and G-protein activator) in a dose-dependent manner reduced the isoflurane block; 120 microm GTPgammaS completely eliminated the block of 0.3 mm isoflurane and reduced the apparent isoflurane potency by approximately 2.4 times (IC50 approximately 0.68 mm). Pretreatment with pertussis toxin or cholera toxin did not eliminate the GTPgammaS-induced protection against the isoflurane block. Furthermore, isoflurane reduced the magnitude of voltage-dependent G-protein-mediated inhibition of N-type Ba2+ currents, and this effect was eliminated by pretreatment with pertussis toxin or cholera toxin. CONCLUSIONS: It was found that activation of G-proteins in a neuronal environment dramatically reduced the isoflurane potency for inhibiting N-type Ba2+ currents and, in turn, isoflurane affected the G-protein regulation of N-type Ba2+ currents. PMID- 12883411 TI - Anesthetic preconditioning: effects on latency to ischemic injury in isolated hearts. AB - BACKGROUND: Anesthetic preconditioning (APC) is protective for several aspects of cardiac function and structure, including left ventricular pressure, coronary flow, and infarction. APC may be protective, however, only if the duration of ischemia is within a certain, as yet undefined range. Brief ischemia causes minimal injury, and APC would be expected to provide little benefit. Conversely, very prolonged ischemia would ultimately cause serious injury with or without APC. Previous investigations used a constant ischemic time as the independent variable to assess ischemia-induced changes in dependent functional and structural variables. The purpose of the study was to define the critical limits of efficacy of APC by varying ischemic time. METHODS: Guinea pig hearts (Langendorff preparation; n = 96) underwent pretreatment with sevoflurane (APC) or no treatment (control), before global ischemia and 120 min reperfusion. Ischemia durations were 20, 25, 30, 35, 40, and 45 min. RESULTS: At 120 min reperfusion, developed (systolic-diastolic) left ventricular pressure was increased by APC compared with control for ischemia durations of 25-40 min. Infarction was decreased by APC for ischemia durations of 25-40 min, but not 20 or 45 min. APC improved coronary flow and vasodilator responses for all ischemia durations longer than 25 min, and decreased ventricular fibrillation on reperfusion for ischemia durations longer than 30 min. CONCLUSIONS: Although APC protects against vascular dysfunction and dysrhythmias after prolonged ischemia, protection against contractile dysfunction and infarction in this model is restricted to a range of ischemia durations of 25-40 min. These results suggest that APC may be effective in a subset of patients who have cardiac ischemia of intermediate duration. PMID- 12883413 TI - Embolism bubble adhesion force in excised perfused microvessels. AB - BACKGROUND: The mechanics of gas embolism bubble adhesion to the vessel wall is poorly understood. New strategies to treat gas embolism may result from an understanding of adhesion forces, including the molecular determinants of bubble adhesion. The authors conducted experiments to measure the adhesion force of bubbles contacting the vessel wall. METHODS: Microbubbles were injected into excised arterioles. Bubbles resided for 5, 10, 20, or 30 min with the endothelium intact or damaged and with a physiologic salt solution, physiologic salt solution with 5% bovine serum albumin, or rat serum as the perfusate. Inflow pressure was raised until the bubble dislodged. The differential pressure across the microbubble, deltaP, was recorded at the moment of bubble movement. Bubble diameter, D, and length, L, were determined by videomicroscopy. The adhesion force per unit surface area of a bubble contacting the vessel wall, K = deltaPD/4 L, was calculated for each experiment. RESULTS: K at 10 min contact time (physiologic salt solution, 141 +/- 29; serum, 153 +/- 57 dyne/cm2) was higher than at 5 min (physiologic salt solution, 56 +/- 22; serum, 71 +/- 29 dyne/cm2), 20 min (physiologic salt solution, 46 +/- 29) and 30 min (physiologic salt solution, 14 +/- 5) (P < 0.05). Endothelium removal reduced K at 10 min (physiologic salt solution, 68 +/- 46; serum, 60 +/- 14 dyne/cm2) (P < 0.05). K was higher with 5% bovine serum albumin present at 10 min (349 +/- 149, P < 0.05), correlating with in vivo estimates. CONCLUSIONS: The adhesion force developed between a microbubble and the vessel wall depends on multiple factors, including bubble residence time, presence of the endothelium, and perfusion solution. PMID- 12883414 TI - The influence of hemorrhagic shock on propofol: a pharmacokinetic and pharmacodynamic analysis. AB - BACKGROUND: Propofol is a common sedative hypnotic for the induction and maintenance of anesthesia. Clinicians typically moderate the dose of propofol or choose a different sedative hypnotic in the setting of severe intravascular volume depletion. Previous work has established that hemorrhagic shock influences both the pharmacokinetics and pharmacodynamics of propofol in the rat. To investigate this further, the authors studied the influence of hemorrhagic shock on the pharmacology of propofol in a swine isobaric hemorrhage model. METHODS: After approval from the Animal Care Committee, 16 swine were randomly assigned to control and shock groups. The shock group was bled to a mean arterial blood pressure of 50 mmHg over a 20-min period and held there by further blood removal until 30 ml/kg of blood was removed. Propofol 200 microg. kg(-1). min(-1) was infused for 10 min to both groups. Arterial samples (15 from each animal) were collected at frequent intervals until 180 min after the infusion began and analyzed to determine drug concentration. Pharmacokinetic parameters for each group were estimated using a three-compartment model. The electroencephalogram Bispectral Index Scale was used as a measure of drug effect. The pharmacodynamics were characterized using a sigmoid inhibitory maximal effect model. RESULTS: The raw data demonstrated higher plasma propofol levels in the shock group. The pharmacokinetic analysis revealed slower intercompartmental clearances in the shock group. Hemorrhagic shock shifted the concentration effect relationship to the left, demonstrating a 2.7-fold decrease in the effect site concentration required to achieve 50% of the maximal effect in the Bispectral Index Scale. CONCLUSIONS: Hemorrhagic shock altered the pharmacokinetics and pharmacodynamics of propofol. Changes in intercompartmental clearances and an increase in the potency of propofol suggest that less propofol would be required to achieve a desired drug effect during hemorrhagic shock. PMID- 12883415 TI - Reactive oxygen species precede the epsilon isoform of protein kinase C in the anesthetic preconditioning signaling cascade. AB - BACKGROUND: Protein kinase C (PKC) and reactive oxygen species (ROS) are known to have a role in anesthetic preconditioning (APC). Cardiac preconditioning by triggers other than volatile anesthetics, such as opioids or brief ischemia, is known to be isoform selective, but the isoform required for APC is not known. The authors aimed to identify the PKC isoform that is involved in APC and to elucidate the relative positions of PKC activation and ROS formation in the APC signaling cascade. METHODS: Isolated guinea pig hearts were subjected to 30 min of ischemia and 120 min of reperfusion. Before ischemia, hearts were either untreated or treated with sevoflurane (APC) in the absence or presence of the nonspecific PKC inhibitor chelerythrine, the PKC-delta inhibitor PP101, or the PKC-epsilon inhibitor PP149. Spectrofluorometry and the fluorescent probes dihydroethidium were used to measure intracellular ROS, and effluent dityrosine as used to measure extracellular ROS release. RESULTS: Previous sevoflurane exposure protected the heart against ischemia-reperfusion injury, as previously described. Chelerythrine or PP149 abolished protection, but PP101 did not. ROS formation was observed during sevoflurane exposure and was not altered by any of the PKC inhibitors. CONCLUSIONS: APC is mediated by PKC-epsilon but not by PKC delta. Furthermore, PKC activation probably occurs downstream of ROS generation in the APC signaling cascade. PMID- 12883416 TI - Brachial plexus examination and localization using ultrasound and electrical stimulation: a volunteer study. AB - BACKGROUND: Current techniques of brachial plexus block are "blind," and nerve localization can be frustrating and time consuming. Previous studies on ultrasound-assisted brachial plexus blocks are mostly performed with scanning probes of 10 MHz or less. The authors tested the usefulness of a state-of-the art, high-resolution ultrasound probe (up to 12 MHz) in identifying the brachial plexus in five locations of the upper extremity and in guiding needle advancement to target before nerve stimulation. METHODS: In this prospective observational study, 15 volunteers underwent brachial plexus examination using an L12-L5 MHz probe and a Philips-ATL 5000 ultrasound unit in the interscalene, supraclavicular, infraclavicular, axillary, and midhumeral regions. Thereafter, an insulated block needle was advanced under direct ultrasound guidance to target nerves before confirmation by electrical nerve stimulation in five volunteers in each of the interscalene, supraclavicular, and axillary regions. The quality of brachial plexus images, anatomic variations, and the technique of needle advancement for nerve localization were recorded. RESULTS: The brachial plexus components were successfully identified in the transverse view as round to oval hypoechoic structures with small internal punctuate echos in all regions examined except the infraclavicular area (visualized in 27% of the cases). The authors' technique of advancing the needle in-line with the ultrasound beam allowed moment by-moment observation of the needle shaft and tip movement at the time of nerve localization. Hypoechoic structures were stimulated electrically and confirmed to be nerves. CONCLUSIONS: These preliminary data show that the high-resolution L12 L5 probe provides good quality brachial plexus ultrasound images in the superficial locations i.e., the interscalene, supraclavicular, axillary, and midhumeral regions. The needle technique described here for ultrasound-assisted nerve localization provides real-time guidance and is potentially valuable for brachial plexus blocks. PMID- 12883417 TI - Evaluation of the lateral modified approach for continuous interscalene block after shoulder surgery. AB - BACKGROUND: Continuous interscalene block is the technique of choice for postoperative pain relief treatment after shoulder surgery. The authors prospectively evaluated the modified lateral approach for the performance of the interscalene catheter block and monitored 700 patients for clinical efficacy and complications during the first 6 months after placement of the catheter. METHODS: A total of 700 adults scheduled to undergo elective shoulder surgery performed with an interscalene brachial plexus block through an interscalene catheter were included in this study. The interscalene brachial plexus block procedure was standardized for all patients. Difficulties in placement of the catheter, clinical efficacy of anesthesia and analgesia, patient satisfaction, and acute and chronic complications were recorded. Patients were observed daily for 5 days for any complications and were evaluated at 1, 3, and 6 months after surgery. Persistence of neurologic complication was investigated by electroneuromyography. RESULTS: A total of 700 adults completed the study. Easy placement of the catheter (one attempt) was achieved in 86% of the patients. Resistance to thread the catheter was encountered in 6%; no major complications were observed during injection of the initial bolus. The success rate for anesthesia was 97%. Postoperative analgesia was efficient in 99%. The concentration and the rate of infusion of ropivacaine had to be increased in 31 patients (6%). In five patients (0.7%), signs of local infection around the puncture point were noted; in one patient (0.1%), a collection of pus was surgically drained. Patient satisfaction was 9.6 on a scale of 0-10. Minor neurologic complications (paresthesias, dysesthesias, pain not related to surgery) were observed in 2.4%, 0.3%, and 0% at 1, 3, and 6 months, respectively. At 1 month, three sulcus ulnaris syndromes, one carpal tunnel syndrome, and one complex regional pain syndrome were diagnosed. Two patients (0.2%) had sensory-motor deficit, which necessitated 19 and 28 weeks to recover. Electromyography was suggestive of partial axonotmesis. CONCLUSION: The lateral modified approach provides good conditions for placement of the interscalene catheter. Anesthesia and analgesia performed through the catheter are efficient. The rates of infection and neurologic complications are low, and patient satisfaction is high. PMID- 12883418 TI - Age-dependent responses to thermal hyperalgesia and mechanical allodynia in a rat model of acute postoperative pain. AB - BACKGROUND: Developmental differences in short- and long-term responses to pain, especially surgical pain, have received minimal attention. The purpose of the present study was to examine postoperative responses in rats of developmental ages paralleling the infant to young adult human. METHODS: The withdrawal threshold to von Frey filament testing and withdrawal latency to hind-paw radiant heating were determined before and for various times after hind-paw incision in rats 2, 4, and 16 weeks of age. Control rats of these ages were observed serially without surgery. RESULTS: In control animals, younger rats were more sensitive to mechanical stimulation and less sensitive to thermal stimulation. Paw incision resulted in similar changes to both types of stimulation in all age groups, peaking 4 h after surgery. However, the return to normal sensitivity to mechanical stimulation, as measured by return of threshold to 80% of normal, occurred more quickly in 2-week-old than in 4- and 16-week-old animals. In contrast, there was no age difference for time to return to normal sensitivity to thermal stimulation after surgery. CONCLUSIONS: The more rapid recovery of the younger animals from the mechanical allodynia but not thermal hypersensitivity after surgery suggests the presence of developmental differences in modulation of A-fiber sensitization after surgery. However, the lack of age difference in recovery of thermal hypersensitivity after surgery suggests that sensitization of C-fiber input has a similar time course of resolution of pain over the ages studied in this model. The neural bases for these developmental differences are under study and may lead to a better understanding of pain during development and altered approaches to treatment of postoperative pain in neonates and infants. PMID- 12883419 TI - Interaction between the spinal melanocortin and opioid systems in a rat model of neuropathic pain. AB - BACKGROUND: The authors recently demonstrated that administration of the melanocortin-4 receptor antagonist SHU9119 decreased neuropathic pain symptoms in rats with a sciatic chronic constriction injury. The authors hypothesised that there is a balance between tonic pronociceptive effects of the spinal melanocortin system and tonic antinociceptive effects of the spinal opioid system. Therefore, they investigated a possible interaction between these two systems and tested whether opioid effectiveness could be increased through modulation of the spinal melanocortin system activity. METHODS: In chronic constriction injury rats, melanocortin and opioid receptor ligands were administered through a lumbar spinal catheter, and their effects on mechanical allodynia were assessed by von Frey probing. RESULTS: Naloxone (10-100 microg) dose-dependently increased allodynia (percent of maximum possible effect of -67 +/- 9%), which is in agreement with a tonic antinociceptive effect of the opioid system. SHU9119 decreased allodynia (percent of maximum possible effect of 60 +/- 13%), and this effect could be blocked by a low dose of naloxone (0.1 microg), which by itself had no effect on withdrawal thresholds. Morphine (1-10 microg) dose-dependently decreased allodynia (percent of maximum possible effect of 73 +/ 14% with the highest dose tested). When 0.5 microg SHU9119 (percent of maximum possible effect of 47 +/- 14%) was given 15 min before morphine, there was an additive antiallodynic effect of both compounds. CONCLUSIONS: Together, these data confirm that there is an interaction between the spinal melanocortin and opioid systems and that combined treatment with melanocortin-4 receptor antagonists and opioids might possibly contribute to the treatment of neuropathic pain. PMID- 12883420 TI - Epidural, cerebrospinal fluid, and plasma pharmacokinetics of epidural opioids (part 1): differences among opioids. AB - BACKGROUND: The pharmacokinetics of epidurally administered drugs has been the subject of many studies, yet drug concentration in the epidural space has never been measured. This study was undertaken to characterize the epidural, cerebrospinal fluid, and plasma pharmacokinetics of epidurally administered opioids on the basis of measurement of drug concentration in each of these compartments after epidural administration. METHODS: Morphine plus alfentanil, fentanyl, or sufentanil were administered epidurally in anesthetized pigs. Microdialysis was used to sample the epidural space and the cerebrospinal fluid for measurement of opioid concentration over time. Plasma samples were obtained from the central venous plasma and the epidural venous plasma. These data were used to calculate relevant pharmacokinetic parameters, including mean residence time, elimination half-lives, areas under the concentration versus time curves, clearance, and volume of distribution for each opioid in each compartment. RESULTS: Some of the more important findings were that the cerebrospinal fluid and plasma pharmacokinetics of the opioids did not parallel their epidural pharmacokinetics and that their hydrophobic character governed multiple aspects of their lumbar epidural pharmacokinetics. CONCLUSIONS: The findings indicate that the spinal pharmacokinetics of these drugs are complex and, in some ways, counterintuitive. Also, the bioavailability of opioids in the cerebrospinal fluid and epidural space is determined primarily by their hydrophobicity, with less hydrophobic drugs having greater bioavailability. PMID- 12883422 TI - Pronociceptive actions of isoflurane: a protective role for estrogen. AB - BACKGROUND: Low concentrations of inhaled anesthetics present in the early postoperative period can increase pain sensitivity. Changes in pain threshold associated with inhaled anesthetics have been reported in male mice, rats, and humans. METHODS: The authors compared the pain-enhancing effects of isoflurane in male and female mice in response to a thermal stimulus and studied the consequences of hormonal manipulation. RESULTS: Isoflurane produced a larger increase in pain sensitivity in female mice. Both castration and oophorectomy resulted in an increase in baseline pain sensitivity and potentiated pain enhancement by isoflurane. At stages of the estrus cycle when estrogen was low, female mice showed greater pain enhancement from isoflurane than at high estrogen stages. Treatment with exogenous estrogen reduced isoflurane-induced pain sensitivity. Exogenous testosterone treatment had a similar effect, which did not occur when enzymatic conversion to estrogen was prevented. CONCLUSIONS: Because both estrogen and testosterone reduce the pronociceptive action of isoflurane, intact females may exhibit a larger increase in pain sensitivity because of their cyclical estrogen levels. Testosterone is effective in the male because of conversion to estrogen. Enhanced pain sensitivity is clearly undesirable in the postoperative setting. If these findings also apply to humans, the menstrual cycle may be an important factor in determining pain levels after emergence from general anesthesia. PMID- 12883421 TI - Epidural, cerebrospinal fluid, and plasma pharmacokinetics of epidural opioids (part 2): effect of epinephrine. AB - BACKGROUND: The ability of epinephrine to improve the efficacy of epidurally administered drugs is assumed to result from local vasoconstriction and a consequent decrease in drug clearance. However, because drug concentration in the epidural space has never been measured, our understanding of the effect of epinephrine on epidural pharmacokinetics is incomplete. This study was designed to characterize the effect of epinephrine on the epidural, cerebrospinal fluid, and plasma pharmacokinetics of epidurally administered opioids. METHODS: Morphine plus alfentanil, fentanyl, or sufentanil was administered epidurally with and without epinephrine (1:200,000) to pigs. Opioid concentration was subsequently measured in the epidural space, central venous plasma, and epidural venous plasma, and these data were used to calculate relevant pharmacokinetic parameters. RESULTS: The pharmacokinetic effects of epinephrine varied by opioid and by sampling site. For example, in the lumbar epidural space, epinephrine increased the mean residence time of morphine but decreased that of fentanyl and sufentanil. Epinephrine had no effect on the terminal elimination half-life of morphine in the epidural space, but it decreased that of fentanyl and sufentanil. In contrast, in the lumbar intrathecal space, epinephrine had no effect on the pharmacokinetics of alfentanil, fentanyl, or sufentanil, but it increased the area under the concentration-time curve of morphine and decreased its elimination half-life. CONCLUSIONS: The findings indicate that the effects of epinephrine on the spinal pharmacokinetics of these opioids are complex and often antithetical across compartments and opioids. In addition, the data clearly indicate that the pharmacokinetic effects of epinephrine in spinal "compartments" cannot be predicted from measurements of drug concentration in plasma, as has been assumed for decades. PMID- 12883423 TI - Use of discharge abstract databases to differentiate among pediatric hospitals based on operative procedures: surgery in infants and young children in the state of Iowa. AB - INTRODUCTION: A pediatric hospital may aim to show governmental agencies, charitable organizations, and philanthropic individuals how its clinical services differ from those of nonpediatric surgical facilities and of other pediatric hospitals. Yet, it is unknown how to use existing databases to quantify where infants and young children undergo surgery, and to use that information to differentiate among facilities. METHODS: Discharge abstracts were used to study inpatient and outpatient operative procedures performed between January and June 2001 in children 0-2 yr old at hospitals or hospital-affiliated outpatient surgery centers in Iowa. RESULTS: Of the 93 facilities performing at least one procedure, the 90 performing 15 or fewer different types of procedures provided surgical care for 80% of procedures. Among procedures performed at these 90 facilities, less than 0.15% were physiologically complex (more than seven American Society of Anesthesiologists' basic units). In contrast, at the larger and smaller pediatric hospitals, the percentages were 26% and 7%, respectively. These pediatric hospitals performed 181 and 73 different types of procedures, respectively; 64% of the physiologically complex procedures performed statewide were performed at the larger pediatric hospital. The smaller pediatric hospital was no more similar to the larger pediatric hospital in its relative volumes of each type of procedure than it was to the other 91 facilities. CONCLUSIONS: Statewide discharge abstract data can be used by a hospital to quantify how its surgical practice differs from that of other hospitals (e.g., to show that it provides a more diverse, comprehensive, and physiologically complex selection of procedures in younger patients). PMID- 12883424 TI - Does the routine prophylactic use of antiemetics affect the incidence of postdischarge nausea and vomiting following ambulatory surgery?: A systematic review of randomized controlled trials. PMID- 12883425 TI - An American contribution to German anesthesia. PMID- 12883426 TI - Sacral osteomyelitis after single-shot epidural anesthesia via the caudal approach in a child. PMID- 12883427 TI - Albuterol-induced lactic acidosis. PMID- 12883428 TI - Is more information better? Intraoperative recall with a Bispectral Index monitor in place. PMID- 12883429 TI - Cortical blindness and neurologic injury complicating cervical transforaminal injection for cervical radiculopathy. PMID- 12883430 TI - Significance of a tourniquet to prevent thromboembolism during total knee arthroplasty as assessed by transesophageal echocardiography. PMID- 12883431 TI - Limitation of supervision. PMID- 12883432 TI - Preoperative epidural placement in elderly patients with hip fractures: a request for essential information. PMID- 12883433 TI - Endotracheal tube foreign body after intubation with a Vital Signs, Inc., lightwand. PMID- 12883434 TI - Air-locked epidural filter. PMID- 12883435 TI - Preoperative antibiotic prophylaxis: the role of the anesthesiologist. PMID- 12883436 TI - Cardioprotective effects of volatile anesthetics in cardiac surgery. PMID- 12883445 TI - Intra-renal resistance reflects warm ischaemic damage, and is further increased by static cold storage: a model of non-heart-beating donor kidneys. AB - BACKGROUND: Non-heart beating donor kidneys increase transplant activity, but their use is associated with a higher rate of both primary non-function and delayed graft function than cadaveric kidneys, due to a period of cold ischaemic damage superimposed on a period of warm ischaemia. We aimed to measure intra renal resistance in machine perfused porcine kidneys subjected to different periods of warm ischaemic injury, with additional, varying, cold ischaemic times, in an attempt to mimic the injury suffered by NHBD kidneys and test the predictive power of viability testing. MATERIAL/METHODS: Landrace pigs were killed by lethal injection, and the kidneys were subjected to varying WITs of 10 90 minutes prior to explantation. Kidneys were subsequently stored for varying cold times of 2 to 48 hours. IRR (pressure/flow) was calculated during 6 hours cold pulsatile machine perfusion. RESULTS: For all WITs, IRR was higher at the start than at the end of machine perfusion (P<0.001). There was a strong correlation between IRR on MP, and WIT, but no correlation after 6 hours MP. Intra-renal resistance increased as kidneys were exposed to longer CITs; this effect was most marked for the longer WITs (P<0.004), However, the slope gradient was similar for the different WITs. CONCLUSIONS: Early IRR accurately reflects kidney in-situ WIT, and machine perfusion reduces IRR whilst cold ischaemia imposed on periods of warm ischaemia increases IRR and attenuates the beneficial effect of MP. Machine perfusion may partially ameliorate the effects of WIT in terms of IRR, and may prove useful in pre-transplant viability assessment of NHBD kidneys. PMID- 12883446 TI - The effect of nitric oxide on the permeability of nasal epithelial cells from healthy and asthmatic donors. AB - BACKGROUND: To see if the higher levels of nitric oxide expired by asthmatics compared to healthy subjects might be of significance to airway function, the effect of nitric oxide and its second messenger, guanosine 3', 5'- cyclic monophosphate (cGMP), on the permeability of human nasal epithelial cells was studied. MATERIAL/METHODS: Cells from healthy and asthmatic donors, collected by swab biopsy, were plated on agar gel before being impaled with a microelectrode to measure their intracellular potential and membrane resistance. RESULTS: Exposure of cells to 300 mM sodium nitroprusside, a nitric oxide donor, caused a profound fall in both parameters in cells from non-asthmatics but no change in cells from asthmatic subjects. A similar response was seen when cells were exposed to 0.9 mM of the permeable form of cGMP, 8-Br-cGMP. Selective inhibition of ion transport pathways in healthy cells indicated that nitric oxide produced changes in permeability consistent with secretion of anions by the cells. CONCLUSIONS: Since anion secretion is associated with fluid secretion in the intact epithelium, we suggest that nitric oxide mediates a protective mechanism to remove foreign material from the airway surface. The defective response to nitric oxide seen in asthmatic cells may contribute to the disease by compromising the removal of allergens from the airway. PMID- 12883447 TI - Production of high-molecular-weight ribonuclease Bsn from the recombinant strain of Bacillus subtilis. AB - BACKGROUND: Ribonucleases (RNases) can be used in both basic and clinical sciences, e.g. in research on developmental processes or on antiviral and antitumor therapy. RNases have great potential as therapeutic entities. On the basis of new ribonucleases new medications can be created. Bacilli synthesize two types of secretory ribonucleases, the well-studied low-molecular-weight ribonucleases and high-molecular-weight ribonucleases. Only two RNases of the second type have so far been described: RNase Bsn from B. subtilis and binase II from B. intermedius. MATERIAL/METHODS: The activity of ribonucleases was determined from the amount of the acid-soluble products of RNA hydrolysis. The cultivation media were optimized for maximum RNase production in terms of the experimental factorial design B2 using BIOPT software. RESULTS: Our investigation of a novel secretory ribonuclease, the Bacillus subtilis RNase Bsn expressed in the recombinant B. subtilis strain 168, showed that it is synthesized in the growth retardation phase, when inorganic phosphate is exhausted in the medium. The biosynthesis of Bsn was found to be suppressed by inorganic phosphate in the medium and activated by small amounts of the transcriptional inhibitor actinomycin D. CONCLUSIONS: Our results show that the biosynthesis of the novel secretory ribonuclease Bsn in recombinant strain Bacillus subtilis 168 is subject to negative regulation by inorganic phosphate, and is activated by small doses of actinomycin D. The stimulating effect of this antibiotic is well pronounced during the active synthesis of ribonucleases, but insignificant when ribonuclease synthesis is inhibited by Pi. PMID- 12883448 TI - Von Willebrand factor in type 1 diabetes: its production and coronary artery calcification. AB - BACKGROUND: Von Willebrand factor (vWF) has generally been regarded as a good predictor of vascular risk. However, no previous studies have examined its relationship with coronary calcification. The aim of this study was to determine whether vWF activity is higher in type 1 diabetic patients than controls; its relationship with cardiovascular risk factors; and to endothelial nitric oxide production and coronary artery calcification. MATERIAL/METHODS: Von Willebrand factor activity was measured in 181 type 1 diabetic patients and 188 controls. Coronary artery calcification was measured by Electron Beam Computed Tomography. Forearm blood flow was measured by venous plethysmography in response to intra brachial infusion of bradykinin, glyceryl trinitrate, noradrenaline and NG monomethyl-L-arginine (L-NMMA) in 149 subjects. RESULTS: Von Willebrand factor was significantly increased in diabetic patients compared to controls (median 100% vs 87%, p=0.001). Von Willebrand factor activity was significantly higher in diabetic patients with micro/macroalbuminuria than those with normoalbuminuria (109% vs 93%, p<0.001). Among diabetic subjects, being in the top quartile for vWF was associated with a lower response to L-NMMA (p=0.009). There was no association between vWF activity and coronary artery calcification in either the diabetic (p=0.9) or control group (p=0.8). CONCLUSIONS: Cardiovascular risk factors including albuminuria do not explain the high vWF activity in type 1 diabetic patients. There is some evidence that vWF correlates with endothelial nitric oxide production. The lack of correlation with coronary artery calcification indicates that vWF is not a useful marker of atheroma burden. PMID- 12883449 TI - Nitric oxide production and intensity of free radical processes in young men with high normal and hypertensive blood pressure. AB - BACKGROUND: Young individuals with high normal blood pressure (HNBP) are at risk for hypertension. The aim of our study was to compare NO production and the intensity of free radical processes in young males with different BP levels. MATERIAL/METHODS: Male subjects aged 18-45 years with normal BP, HNBP, and hypertension underwent physical and cardiological examination. NO production was evaluated by measuring plasma nitrite and nitrate (NOx) and 24-hour urinary excretion of NOx. Lipid peroxidation (LP) intensity and serum antioxidant activity (AOA) were measured using the biochemiluminescence method. RESULTS: HNBP was associated with increased 24-hour systolic BP (SBP), diastolic BP (DBP), SBP and DBP variability, plasma and 24-hour urinary NOx and LP intensity, and decreased total AOA as compared to normotensive controls. We observed a direct nonlinear correlation between plasma NOx and SBP and between 24-hour urinary NOx and DBP, and a close inverse correlation between LP intensity and AOA in patients with HNBP. In the presence of two cardiovascular risk factors (smoking and obesity), patients with HNBP displayed higher LP intensity and lower levels of NOx than in both nonsmokers with normal body weight and control subjects. In hypertensive patients, SBP, DBP and LP intensity inversely correlated with plasma and urinary NOx. CONCLUSIONS: Activation of LP processes and depression of AOA proceed in parallel with declining NO production and severity of hypertension. Early correction of the revealed disorders before the appearance of clinical symptoms may be promising in terms of prevention and treatment of cardiovascular disease. PMID- 12883450 TI - Blood serum and neutrophil L-arginine concentrations and nitric oxide release by neutrophils in chronic uremic patients and healthy persons. AB - BACKGROUND: Plasma levels of L-arginine have previously been evaluated in several studies to reveal either down- or up-regulation of the L-arginine/NO pathway in chronic uremia. MATERIAL/METHODS: We studied L-arginine plasma levels along with intracellular neutrophil concentration and NO release by peripheral blood neutrophils in patients with chronic uremia. RESULTS: The L-arginine plasma concentration was found to be significantly higher (64.4 +/- 12.0 micromol/L) in chronic uremic patients (n=25) than in healthy controls (33.0? +/- 10.0 micromol/L; n=25). Neutrophil homogenate L-arginine levels were substantially increased in uremic patients (7.95? +/- 1.10 nmol/10(8) cells) as compared with controls (5.22 +/- 0.46 nmol/10(8) cells). The in vitro release of NO by unstimulated neutrophils was lower in uremic patients (0.14? +/- 0.05? micromol s(-1)) than in healthy persons (0.48 +/- 0.20 micromol s-1). The NO release after uremic neutrophil stimulation with 10(-6) M fMLP was 21.42 +/- 2.13 micromol s( 1), while after PMA it was 31.01 +/- 1.99 micromol s(-1). NO release after normal neutrophil stimulation with 10(-7) M fMLP was 19.52 +/- 2.32 micromol s(-1), and after PMA was 28.63 +/- 3.06 micromol s(-1). CONCLUSIONS: In chronic uremic patients plasma and neutrophil concentrations of L-arginine were significantly higher. However, there were no significant differences in NO release between normal and uremic neutrophils after stimulation. Our findings may have implications for the impact of the L-arginine-nitric oxide signaling pathway on the pathophysiological changes in the L-arginine: NO pathway seen in chronic uremia. PMID- 12883451 TI - A survey of combined epidural-propofol anesthesia with noninvasive positive pressure ventilation as a minimally invasive anesthetic protocol. AB - BACKGROUND: Combined epidural-propofol anesthesia with use of noninvasive positive pressure ventilation (NPPV) via the nose has been used routinely in our operating theaters. The purpose of this report was to present a survey of this anesthesia. MATERIAL/METHODS: 265 adult patients undergoing lower extremity or lower abdominal gynecological surgery during 1999 were examined. After epidural anesthesia, patients were given propofol infusion. NPPV was applied with an inspiratory/expiratory positive airway pressure of 14/8 cm H2O, a respiratory rate of 10 breaths/min, and oxygen delivery into the nasal mask resulting in a concentration of 40% or an inspiratory oxygen fraction of 0.35. Epidural anesthesia was continuously applied after surgery for postoperative pain relief. Various data related to the surgery or anesthesia were evaluated both on the day of surgery and on postoperative day 1. RESULTS: Of 265 patients, 3 patients could not receive our anesthetic protocol. Of the residual 262 patients, no patients showed serious clinical problems during anesthesia, excluding for hypotension, which was observed in 31-56% patients and was treated with ephedrine injection. Patients informed us of good analgesia (98%), feelings (78%) and dreams (47%). On postoperative day 1, postoperative analgesia and mood conditions were satisfactory. There were no patients complaining of intraoperative awareness. CONCLUSIONS: The principle of our anesthesia consists of epidural anesthesia, sole propofol infusion and noninvasive airway management, so as to provide an anesthetic technique with minimal invasiveness. Although airway maintenance by NPPV is not always suitable, our anesthesia is practicable for certain kinds of operations. PMID- 12883452 TI - Clinical outcome in monosegmental fusion of degenerative lumbar instabilities:instrumented versus non-instrumented. AB - BACKGROUND: Degenerative lumbar instabilities are one of the major reasons for low back pain, especially in elderly people. There is still controversy in the literature about the clinical outcome of instrumented monosegmental fusions compared to noninstrumented posterolateral fusions. MATERIAL/METHODS: In this study we described the postoperative outcome of 33 patients who underwent posterolateral monosegmental spinal fusion:instrumented (17 patients)or noninstrumented (16 patients).All patients received the same instrumentation system. Medical history (considering daily activities,sports,social contacts and quality of life),clinical examination and radiological analysis served as evaluation parameters. Data were collected by a questionnaire based on the Oswestry Disability Score for low back pain. RESULTS: 86.6%of all subjects showed an improvement of postoperative health status. Although patients with fusion without internal fixation needed a reduced dose of postoperative analgesics compared to the monosegmental non-instrumented group,there were no statistical differences in patient-reported,clinical and radiographical outcome between the two groups. CONCLUSIONS: The results do not indicate a benefit in outcome from added instrumentation in elective lumbar fusions. This should be considered in each single case when a spinal fusion in the treatment of degenerative lumbar instability is indicated. PMID- 12883453 TI - Screening for metabolic syndrome in hypertensive and/or obese subjects registered in primary health care in Hungary. AB - BACKGROUND: The screening of subjects at risk for metabolic syndrome is of considerable importance in order to prevent atherosclerosis. MATERIAL/METHODS: In order to assess the clinical-laboratory characteristics of metabolic syndrome,a screening procedure was performed in subjects (age 20 -65)who exhibited hypertension and/or abnormal body mass index (BMI)and/or elevated waist-hip ratio,excluding patients with known diabetes. An oral glucose tolerance test with 75 g glucose was performed. Plasma glucose and insulin values were measured,as well as plasma lipids. RESULTS: In subjects available for complete statistical analysis (n=944;women/men ratio 1.37:1),hyperinsulinemia was detected in 52.9%. Hyperinsulinemia with normal glucose tolerance was more often detected (33.2%),hyperinsulinemia with impaired glucose tolerance [IGT ]or diabetes less frequently (13.0%and 6.7%,respectively). When abnormal clinical signs were separately analysed,hyperinsulinemia was found in 56.8%of subjects with abnormal BMI,in 43.8% of subjects with abnormal waist-hip ratio,and in 27.1% of subjects with hypertension. Metabolic syndrome (WHO criteria,modified)was diagnosed in 35.2%of subjects,with male predominance (men:40.6%;women:31.2%;p<0.01). CONCLUSIONS: Obesity (abnormal BMI;abnormal waist-hip ratio)is of greater importance than hypertension alone for detecting subjects with hyperinsulinemia.Routine clinical and laboratory investigations (anthropometric data,measuring blood pressure,oral glucose tolerance test)are simple but useful for identifying subjects with metabolic syndrome,enabling the implementation of a primary strategy to prevent cardiovascular morbidity. PMID- 12883454 TI - Photostimulation of coronary arteries with low power laser radiation: preliminary results for a new method in invasive cardiology therapy. AB - BACKGROUND: The main problem after percutaneous coronary intervention (PCI) is restenosis affecting the site where dilatation is performed. In order to minimize its occurrence, the method of intravascular laser photostimulation (LP) with low power irradiation has been developed. The new procedure is carried out during PCI. MATERIAL/METHODS: A special setup was prepared for intravascular photostimulation with 808 nm wavelength laser diode and special diffuser, delivering the laser light into the coronary artery. The construction of the device makes it possible to irradiate the coronary artery in the place of previously performed of dilatation, in an satisfactory and programmable manner and with uniform intensity. We describe our own, unique LP procedure. Two pilot LP procedures carried out in our clinic are described here. The patients were diagnosed before treatment and followed up three and six months after the LP procedure with non invasive tests. After six months, a control angiography was also performed. RESULTS: The procedures were well tolerated. In both cases the follow-up examinations showed no evidence of restenosis. CONCLUSIONS: The new, innovative technique of intravascular low power laser photostimulation is a relatively inexpensive and easy to use treatment method. No negative side effects were observed after two procedures. The results obtained are very promising. Because the group of treated patients is still too small, the results cannot yet be subjected to statistical analysis. PMID- 12883455 TI - Intravenous nitro-glycerine versus general anaesthesia for placental extraction- a sequential comparison. AB - BACKGROUND: Postpartum haemorrhage due to retained placenta is one of the commonest life-threatening conditions during the third stage of labour. Uterine relaxation is usually required to facilitate placental removal. 'Full-stomach' obstetric patients (which includes those who delivered within 48 h), parturients with a history of antepartum or recurrent postpartum hemorrhage, grand multiparity, twin pregnancy, and those with cardiac abnormalities may benefit from an alternative to volatile-based general anaesthesia for uterine relaxation to avoid complications associated with the technique (e.g. aspiration pneumonitis and cardiovascular compromise). CASE REPORT: A 34-year-old gravida 4, para 3 parturient with rheumatic valvular heart disease presented with retained placenta and postpartum haemorrhage on two consecutive deliveries and had the placenta removed manually by the same surgeon under two different anaesthetic techniques. On the first occasion, general anaesthesia was administered whereas only i.v. fentanyl and nitro-glycerine were used on the second occasion. The postoperative course was uneventful on both occasions. CONCLUSIONS: The use of nitro-glycerine was found to be efficacious for manual removal of placenta with minimal haemodynamic perturbations, avoiding the use (and associated risks) of general anaesthesia for uterine relaxation. The ability of nitro-glycerine to reduce spontaneous uterine activity, induce uterine relaxation, coupled with its short duration of action and high efficacy, may render it a safe alternative to general anaesthesia for facilitating intrauterine manoeuvres. Nitro-glycerine may be useful especially in patients with associated co-morbid chronic cardiac conditions, e.g. rheumatic heart disease, which is characterised by impaired haemodynamics and cardiac reserves. PMID- 12883456 TI - Papillary carcinoma of thyroglossal duct cyst. AB - BACKGROUND: Thyroglossal duct cysts are the most common congenital abnormality of thyroid development. They represent the most common midline neck masses in childhood, however, can be found in adults. Less than 1% of these cases are malignant. Our patient was a 28-yr-old man and his chief complaint was a progressively enlarged paramedian neck mass for a duration of 6 months. RESULTS: We describe the imaging and pathologic characteristics of papillary carcinoma of thyroglossal duct cyst in a 28 years old man and subsequent treatment recommendations. The pathologic examination revealed papillary carcinoma of thyroglossal duct cyst followed by a total thyroidectomy. An axial CT scan of the neck showed a left Paramedian cyst,measuring 15 x 18 mm in the submental region. Neck dissection was done followed by 131I ablation. CONCLUSIONS: Base upon our past and current experience,we recommend that thyroglossal duct cyst with a microscopic focus of papillary carcinoma without cyst wall invasion be managed with Sistrunk procedure along with effective suppressant dose of thyroxine. PMID- 12883457 TI - Serum cytokine level and production of reactive oxygen species (ROS) by blood neutrophils from a schizophrenic patient with hypersensitivity to neuroleptics. AB - BACKGROUND: There have been few publications concerning the role of the immune system in neuroleptic intolerance. Some studies have shown that in neuroleptic malignant syndrome (NMS), associated with disseminated intravascular coagulation (DIC), the serum level of tumor necrosis factor alpha (TNF-alpha) increases significantly, which is thought to trigger the onset of DIC. CASE REPORT: A 23 year-old woman suffering from catatonic schizophrenia developed hypersensitivity to neuroleptics. One month before being referred to the present authors, she had a haloperidol-induced NMS episode in another psychiatric hospital, with high temperature, CPK activity, muscle rigidity and leukocytosis. On admission to our clinic and after treatment with promazine, laboratory tests showed an increase in serum CPK activity and mild leukocytosis. Neuroleptic treatment was discontinued, and the serum level of CPK and white blood cell count was monitored daily for 7 days, as well as the serum level of some cytokines and the production of reactive oxygen species (ROS) by blood neutrophils. The serum levels of interleukin 1 alpha (IL-1), IL-6 and TNF-alpha changed significantly over the observation period, forming waves with peak activity of IL-6 and TNF-alpha exceeding normal levels. The level of IL-1 alpha was within the control range. ROS production by the patient's blood neutrophils was also increased, as well as catalase serum activity. CONCLUSIONS: Some proinflammatory cytokines may participate in the mechanisms leading to the development of neuroleptic intolerance in schizophrenic patients. Cytokine-stimulated ROS production may participate in tissue injury and increase CPK serum activity. PMID- 12883458 TI - Rapid and quantitative detection of human septin family Bradeion as a practical diagnostic method of colorectal and urologic cancers. AB - BACKGROUND: Malignant tumor progression is a complex and multi-gene event which can not be easily detected or predicted. The detection of malignant cells using marker genes is hampered by the fact that these markers are only expressed by certain malignancies or lack sensitivity and/or specificity. We have reported a human septin family gene Bradeion, which shows strong cancer-specific expression in colorectal and urologic cancers as a result of carcinogenesis. MATERIAL/METHODS: Diagnostic efficacy and validity of Bradeion gene expression were tested by two independent systems, one is a protein detection method using monoclonal antibody based immuno-chromatographic membrane strip tests (a nitrocellulose test strip assay), and another is a gene expression detection method, quantitative RT-PCR. The technology has been established using Bradeion fusion proteins, in vitro cultivated human cancer cell lines, and also patients' test samples with controls. RESULTS: Bradeion test strip by combination with two monoclonal antibodies are valid for the detection of 1 ng/ml Bradeion, and successfully applied for patient urine samples with no false-positive results. Positive detection rates were over 70% of the patient urine samples so far tested (prostate cancer, renal cell carcinoma, and bladder cancer) in 15 to 30 minutes. Quantitative RT-PCR resulted in significantly high copy numbers of 0.4-3.0-3.0 x 10(5) per microg total RNA in patients' tissue samples, whereas those from normal tissue or other cancers found negative. CONCLUSIONS: The present study introduces the practical diagnostic methods using a disease-specific molecular marker, which provides safe, economical, and rapid clinical screening of cancer. PMID- 12883459 TI - Prenatal assessment of ventriculomegaly: an anatomical study. AB - BACKGROUND: The aim of the study was to analyze the development of the lateral ventricles during the 1st and 2nd trimester of fetal life using computerized image processing, and to compare the findings with the results obtained by ultrasound imaging and MRI. MATERIAL/METHODS: The material consisted of 32 fetuses from spontaneous abortions, 54-235 mm crown-rump length. After detached craniotomy, the brains were cut into axial sections; the sections were filmed with a video camera and then analyzed using specialized software RESULTS: In 12 analyzed brains, no significant pathological changes were observed in the cerebral hemispheres, whereas the remaining 20 (63%) demonstrated visible pathology. In 10 cases there were areas of leukomalacia, in 5 intra- and periventricular hemorrhages, and in 2 fetuses ventriculomegaly with lateral ventricular triangles over 10 mm wide (in cases of active hydrocephalus and colpocephaly). In 1 case of an 18-week-old fetus, lateral ventricular morphology typical of hydrocephalus (generalized distension) was observed with ventricular triangles 8.5 mm wide. The other 2 fetuses demonstrated developmental defects. The frontal horns were the most markedly enlarged in both cases of hydrocephalus (100%) and were semicircular, whereas after intra- and periventricular hemorrhages they were less enlarged and triangular, with the base of the triangle directed to the front and frequent significant asymmetry. CONCLUSIONS: The shape of the ventricular system, including that of the frontal horns, is important in the diagnostics of fetal CNS. PMID- 12883460 TI - Annual radiation background in the City of Isfahan. AB - BACKGROUND: Measurement of environmental exposure is very important from different points of view especially for human health. In Iran, it is measured in some cities especially in high background areas such as Ramsar, but so far there is not any measurement in Isfahan. MATERIAL/METHODS: Measurements were performed using CaSO4: Dy TLDs. The locations under investigations were 52 health centers distributed all around Isfahan city. Each TLD badge was put in a special plastic bag and left over the roofs of the selected health center for a month. The procedure was repeated for all 12 months of the year 1379 (21st March 2000 to 20th March 2001). RESULTS: The maximum and the minimum of obtained results for dose equivalent in different months and locations were 15.9 x 10(-2) and 6.5 x 10(-2) mSv. Obtained maximum and minimum of the means between all the locations were 10.5 x 10(-2) and 8.6 x 10(2) mSv for the whole year. Monthly mean and SD for Isfahan city for the whole year were 9.7 x 10(-2) and 1.5 x 10(-2) respectively therefore mean annual dose equivalent in Isfahan city is 1.16 x 0.18 mSv. The results do not show any high background radiation area. CONCLUSIONS: From the total of 624 measurements at 52 sites of Isfahan, obtained annual outdoor X and gamma ray dose equivalent for a person living in isfahan is 1.16 x 0.18 mSv. This figure is about 48.3% of the world average total radiation background reported by UNSCEAR. PMID- 12883461 TI - Occupational risk of hepatitis A infection among health care providers in northeastern Poland. AB - BACKGROUND: Vaccinating health care workers against hepatitis A has been suggested, but it has not been clearly shown that this group is at increased risk for HAV infection. The objective of our study was to evaluate the prevalence of anti-HAV antibodies among health care workers, and to assess the risk factors associated with anti-HAV seropositivity. MATERIAL/METHODS: Total anti-HAV antibodies in serum were determined among 252 health care workers from three hospital departments (internal medicine, infectious diseases and surgery) in Bialystok, in northeastern Poland. The control group consisted of 250 age and sex matched healthy individuals who are not health-care professionals. RESULTS: Anti HAV antibodies were found in 180 of the 252 health care workers (71.4%) and in 153 of the 250 healthy controls (61.2%, p<0. 05)). The difference in anti-HAV prevalence between health care workers and the control group occurred only in individuals up to 40 years of age (59.2% and 43.4% respectively, p<0. 05) while no difference could be seen in those older than 40 years (88.6% and 88.8% respectively). Among health care workers there was no association between anti HAV seropositivity and sex, employment as a physician, nurse or ward attendant, or ward of employment. Health care workers older than 40 years were anti-HAV positive more frequently than younger (88.6 and 59.2% respectively, p<0.05). CONCLUSIONS: In northeastern Poland. younger health care workers ((40 years) may be particularly prone to occupationally acquired hepatitis A infections. PMID- 12883462 TI - Preliminary results with the use of an albumin-glutaraldehyde tissue adhesive in lung surgery. AB - BACKGROUND: The purpose of this study was to test the performance of an albumin glutaraldehyde tissue adhesive, BioGlue(r) Surgical Adhesive (BioGlue) in the sealing of air leaks from pulmonary parenchyma and bronchopleural fistulas. MATERIAL/METHODS: Between March 2000 and November 2001 BioGlue was applied in 38 randomly selected patients, who underwent 39 operations. The mean age was 51.4 years (range 19 to 75 years). A median of 5 cc of BioGlue was used per patient (range 5 to 20 cc). The operations included 36 thoracotomies, 2 video-assisted thoracoscopies and one rigid bronchoscopy. RESULTS: The duration of air leak ranged from 0 to 2 days with a median of 1 day. The duration of total (air and fluid) chest tube drainage ranged from 1 to 12 days with a median of 3 days. Complications were observed in 3 patients (8%) and included atelectasis in one and residual space in 2. Three patients died because of preexisting respiratory failure unrelated to BioGlue application. Hospitalization ranged from 4 to 16 days with a median of 6 days and was prolonged in some patients because of their primary disease (empyema, bronchopleural fistula, etc.). CONCLUSIONS: The use of BioGlue proved to be safe and effective in the sealing of lung lacerations and in preventing air leakage from suture or staple lines in emphysematous lungs. It was also successful in sealing bronchopleural fistulas when applied either intra bronchially through the rigid bronchoscope or during thoracotomy. PMID- 12883463 TI - Somatostatin infusion and hemodynamic changes in patients with non-variceal upper gastrointestinal bleeding: a pilot study. AB - BACKGROUND: Intravenous somatostatin decreases acid secretion, splanchnic blood flow, and portal pressure, but the evidence for its efficacy in the treatment of non-variceal upper gastrointestinal bleeding has been mixed. We aimed to evaluate the vasoactive effect and possible mechanisms of somatostatin infusion in the cessation of non-variceal upper gastrointestinal bleeding. MATERIAL/METHODS: Patients with non-variceal upper gastrointestinal bleeding without portal hypertension were enrolled in the study. They were given somatostatin infusion in a dose of 250 microgr/hour for 72 hours. Superior mesenteric arterial average flow velocity (SMA-V), SMA pulsatility index (SMA-PI), portal venous volume flow (PV-F), and renal artery resistance index (RA-RI) were measured two times for each patient by Doppler ultrasound; once on the first day of infusion therapy and again 6 hours or more after stopping the infusion. RESULTS: 21 patients (12 male, mean age 44.1 +/- 9.9) with bleeding peptic ulcer were enrolled. During somatostatin infusion, PV-F was 33.7 +/- 12.7 cm3/sec. After stopping infusion, it increased to 56.3 +/- 16.0 cm3/sec (p=0.001). SMA-V was 39.7 +/- 13.1 cm/sec and 64.4 +/- 15.1 cm/sec during somatostatin infusion and after cessation of somatostatin respectively (p=0.01). SMA-PI was 2.0 +/- 0.8 during somatostatin infusion but 2.8 +/- 0.8 without somatostatin infusion (p=0.02). However, RA-RI showed no difference between states with or without somatostatin infusion (p>0.05). CONCLUSIONS: Somatostatin infusion causes a decrease in arterial blood flow to the stomach and duodenum in patients with non-variceal upper gastrointestinal bleeding without portal hypertension. Somatostatin therapy also decreases portal blood flow while not altering renal blood. PMID- 12883464 TI - Atorvastatin reduces microalbuminuria in patients with familial hypercholesterolemia and normal glucose tolerance. AB - BACKGROUND: Anecdotal evidence indicates that significant lowering of LDL cholesterol by means of LDL apheresis may reduce the level of microalbuminuria in heterozygous familial hypercholesterolemia. We therefore examined whether treatment with the most potent LDL-lowering drug available (atorvastatin) has a similar effect on the level of microalbuminuria. MATERIAL/METHODS: In a case series, 100 patients with familial heterozygous hypercholesterolemia were started on 10 mg atorvastatin. 62 patients were switched to 40 mg atorvastatin once in the evening when the LDL-cholesterol treatment goals were not reached within 1 month. RESULTS: Baseline serum creatinine clearance significantly improved after 1, 3 and 6 months, while serum urea and serum creatinine were unchanged. Blood pressure exhibited a lowering trend. After one month of treatment, the mean level of microalbuminuria was significantly improved in both dose regimens, showing further improvement after 3 months and stabilizing thereafter. CONCLUSIONS: These data indicate that significant lowering of LDL-cholesterol with atorvastatin may favorably affect kidney function, in particular microalbuminuria as a measure of endothelial function. It remains to be seen whether this effect can be attributed to lipid lowering alone. PMID- 12883465 TI - Role of acyclovir gel in herpes simplex: clinical implications. AB - BACKGROUND: Acyclovir (ACY) is effective in the treatment of herpes simplex (HSV 1) & (HSV-2) but has systemic toxic effects if given orally or intravenously. ACY has not been used to treat the disease topically due to poor drug penetration into skin. A novel 1% liposomal ACY topical gel in a 5% Hydroxypropylmethyl cellulose (HPMC) K4M gel base has been developed and clinically evaluated in HSV 1 and HSV-2 patients. MATERIAL/METHODS: 26 patients suffering from recurrent mild facial (HSV-1) and genital (HSV-2) infections (HSV-1: 4F, 6M, age 21-34 years; HSV-2: 16M, age 24-40 years) were subjected to double blind clinical evaluation. Plain ACY gel (PAG) and Liposomal ACY gel (LAG) were clinically evaluated by application five times daily on herpetic lesions for up to eight weeks. RESULTS: A significant increase in the average percent improvement of lesion healing was observed in HSV-1 and HSV-2 patients after 2-3 weeks treatment with LAG, as well as a significant decrease in side effects associate with ACY, such as itching and burning in both HSV-1 and HSV-2, and burning micturation in HSV-2. CONCLUSIONS: The results demonstrate that a five-fold reduction in the ACY content in liposomal gel is sufficient for the complete healing of herpetic lesions in HSV-1 and HSV-2 infection. The increased duration of topical therapy may be acceptable for patients suffering from mild herpetic lesions because of the advantage of avoiding systemic and local side effects PMID- 12883466 TI - Recent insights into the pathophysiology of paroxysmal nocturnal hemoglobinuria. AB - Paroxysmal nocturnal hemoglobinuria (PNH) is a unique clonal stem cell disorder characterized by intravascular hemolysis, thrombotic events and bone marrow failure. There has been accelerated progress in understanding the mechanisms underlying the clinical features of the disease over the last decade. The development of PNH requires not only a somatic mutation of the phospatidylinositol glycan complementation class A (PIG-A) gene, but also a survival advantage of the PNH clone ('dual pathogenesis' theory). There is increasing evidence that negative selection against the non-mutated cells rather than positive selection of the PIG-A gene mutant cells is responsible for the dominance of the PNH clone. In this review, we summarize the important advances in the understanding of PNH, but we also concentrate on the presence of PNH clones in other hematological disorders, including aplastic anemia (AA), myelodysplastic syndromes (MDS), acute leukemias, and myeloproliferative and lymphoproliferative syndromes. The fuller comprehension of the pathophysiology of PNH may have wider implications than for PNH itself, as indicated by the presence of PNH clones in these hematological malignancies, and by the therapeutic implications of this fact, as already described in patients with AA and MDS. PMID- 12883467 TI - Complications of salvage esophagectomy. AB - Definitive chemoradiation (without surgery) is an accepted treatment for esophageal cancer. Persistent or recurrent local disease is not infrequent after chemoradiation, and this is its greatest drawback. Selected patients with isolated local failures of definitive chemoradiation can be salvaged by esophagectomy. However, salvage esophagectomy is a high-risk operation; operative mortality is higher than for standard esophagectomy. In contrast to planned esophagectomy after induction (neoadjuvant) therapy, salvage esophagectomy is often carried out months after completion of definitive chemoradiation. Previous thoracic radiotherapy contributes to the genesis of postoperative complications. Respiratory failure (ARDS, pneumonia) and septic complications of esophageal reconstruction (anastomotic leaks) are major causes of postoperative death. Gastric conduit necrosis, airway necrosis, and tracheogastric fistulae are infrequent but highly lethal complications. We propose modifications of surgical practice that may reduce complications. These include strict guidelines for patient selection, conservative mediastinal dissection, prevention of intraoperative lung injury (barotrauma and oxygen related toxicity), near total esophagectomy with cervical esophagogastric anastomosis, anterior mediastinal reconstruction, judicious use of staged reconstruction, perioperative enteral nutritional support, and aggressive detection and treatment of postoperative complications. By conceptually breaking the operation into component parts, such as resection and reconstruction, and then modifying each component to minimize complications, we believe that the operative mortality of salvage esophagectomy can PMID- 12883468 TI - Myocardial redox stress and remodeling in metabolic syndrome, type 2 diabetes mellitus, and congestive heart failure. AB - Over the past three decades, we have witnessed an improvement of survival in those patients with the trio of metabolic syndrome, prediabetes, and overt type 2 diabetes mellitus. Revolutionary changes in technology and an improved understanding of the mechanisms involved in acute coronary syndromes have resulted in this observation. Due to advances in coronary care, we are currently at a crossroads, wherein, the mortality from acute cardiovascular events have been declining and the mortality associated with this trio has been increasing due to congestive heart failure (CHF). This intersect between the two causes of death represent a challenge for the future, as the numbers of patients with this deadly trio are undergoing exponential growth not only in the U.S. but also abroad as more countries undergo urbanization and adopt a western-type lifestyle of over nutrition and under exercise. Thus, we live to die another day. There are multiple metabolic toxicities in this toxic trio, which predispose to an increase in reactive oxygen species and resultant redox stress within the vascular intima and myocardium. By aggressively reducing the elevated substrates producing reactive oxygen species we may be able to restore our individual, endogenous, potent, antioxidant (antiredoxidant) network. Appropriately, we need to examine the mechanisms that result in the development and transition from diastolic and systolic dysfunction to the clinical syndrome of overt CHF with its inherent increase in morbidity and mortality. PMID- 12883469 TI - H. pylori infection, atrophic gastritis, cytokines, gastrin, COX-2, PPAR gamma and impaired apoptosis in gastric carcinogenesis. AB - BACKGROUND: Helicobacter pylori (Hp) infection represents a crucial factor in pathogenesis of gastric cancer (GC). Factors emanating from bacterium as well as from environmental contributions such as salt diet and inadequate supply of antioxidants, affect the risk for GC development. RESULTS: Atrophic gastritis is considered to be a precursor lesion of intestinal type GC that is accompanied by hypergastrinemia with subsequent induction of cyclooxygenase-2 (COX-2), whose products are responsible for slowing apoptosis and for angiogenesis in GC tumor. The involvement of proinflammatory cytokines (especially IL-1 and IL-8) and reactive oxygen species (ROS) due to NF kappa B activation, increased cell proliferation combined with inhibition of apoptosis as well as upregulation of peroxisome proliferation activated receptor gamma (PPAR gamma) and inducible nitric oxide synthase (iNOS) appear to be major molecular biology alterations in pathogenesis of GC. CONCLUSIONS: These results suggest the therapeutic usefulness of inhibitors of gastrin expression and release such as powerful somatostatin analogs (Sandostatin) or blockers of COX-2 (coxibs) in the control of GC development and progression as chemopreventive agents. Comparative genomic and proteomic is the key in identifying biomarkers in host and bacterium for the prediction of gastric cancer in Hp-infected patients. PMID- 12883470 TI - Heme oxygenase in liver transplantation: heme catabolism and metabolites in the search of function. PMID- 12883471 TI - Integrin-mediated control of cell growth. PMID- 12883472 TI - Can interferon prolong life? PMID- 12883473 TI - Visualization of early events in tumor formation of eGFP-transfected rat colon cancer cells in liver. AB - Colon cancer preferentially metastasizes to the liver. To determine cellular backgrounds of this preference, we generated an enhanced green fluorescent protein (eGFP)-expressing rat adenocarcinoma cell line (CC531s) that forms metastases in rat liver after administration to the portal vein. Intravital videomicroscopy (IVVM) was used to visualize early events in the development of tumors in livers of live animals from the time of injection of the cancer cells up to 4 days afterward. Based on information obtained with IVVM, tissue areas were selected for further analysis using confocal laser scanning microscopy (CLSM), electron microscopy (EM), and electron tomography. It was shown that initial arrest of colon cancer cells in sinusoids of the liver was due to size restriction. Adhesion of cancer cells to endothelial cells was never found. Instead, endothelial cells retracted rapidly and interactions were observed only between cancer cells and hepatocytes. Tumors developed exclusively intravascularly during the first 4 days. In conclusion, initial steps in the classic metastatic cascade such as adhesion to endothelium and extravasation are not essential for colon cancer metastasis in liver. PMID- 12883474 TI - Mechanism of beta 1-integrin-mediated hepatoma cell growth involves p27 and S phase kinase-associated protein 2. AB - Although cooperative interactions between growth factors and integrins, cell surface receptors for extracellular matrices (ECM), have been reported, little is known about the interaction between hepatocyte growth factor (HGF) and integrin in hepatoma cells. We investigated the effects and mechanisms of integrin on the proliferation of hepatoma cells regulated by HGF. Human HepG2 hepatoma cells stably transfected with beta 1-integrin were treated with HGF and compared with parental and mock-transfected control cells. Cell proliferation and expression of cyclin-dependent kinase (Cdk) inhibitors and S-phase kinase-associated protein 2 (Skp2), were investigated. HGF dose-dependently suppressed the proliferation of parental and mock-transfected HepG2 cells. However, cells overexpressing beta 1 integrin exhibited increased proliferation in response to HGF. Although HGF increased p27 and decreased Skp2 expression in the parental and mock-transfected cells, the p27 and Skp2 levels in cells overexpressing beta 1-integrin were not altered by HGF. Interestingly, HepG2 cells overexpressing beta 1-integrin showed increased Skp2 expression. Furthermore, HGF did not reduce the proliferation of HepG2 cells transfected with antisense p27 or sense Skp2. Thus, HGF suppresses HepG2 cell proliferation by directly increasing p27 expression and indirectly decreasing Skp2 expression, and beta 1-integrin modulates the responsiveness of hepatoma cells to HGF via a p27-dependent manner by increasing Skp2. In conclusion, these results strongly suggest that integrin-mediated signals from the ECM can modulate growth factor-mediated signals in hepatoma cells, and may contribute to the growth of hepatocellular carcinomas. PMID- 12883475 TI - A common set of immediate-early response genes in liver regeneration and hyperplasia. AB - Partial hepatectomy (PH) and some tumor-promoting agents stimulate hepatocyte cell proliferation, but each treatment acts through distinct transcription factors. We compared mouse immediate-early gene expression changes after PH with those induced by 1,4-bis[2-(3,5-dichoropyridyloxy)]benzene (TCPOBOP), a tumor promoting liver mitogen. PH activates nuclear factor kappa B (NF-kappa B) and Stat3, whereas TCPOBOP is a ligand for the nuclear receptor, constitutive androstane receptor (CAR). RNA from 1 and 3 hours after each treatment was hybridized to a 9,000 complementary DNA (cDNA) microarray. Of about 6,000 messenger RNAs that had detectable expression, 127 showed reproducible up regulation or down-regulation at a significant level. The TCPOBOP response was more discrete than the PH response; they amounted to 1% and 1.9% of positive hybridizations, respectively. Twenty-three genes were regulated only by TCPOBOP, 57 only by PH, and 59 by both treatments. More detailed analysis defined 16 clusters with common patterns of expression. These patterns and quantification of hybridization levels on the array were confirmed by Northern blots. TCPOBOP selectively activated expression of a number of detoxification enzymes. In conclusion, the genes that were regulated by both treatments suggest down regulation of apoptosis, altered signal transduction, and early biogenesis of critical cell components. PMID- 12883476 TI - Massive liver growth in mice induced by systemic interleukin 6 administration. AB - The multifunctional cytokine interleukin 6 (IL-6) is expressed in a wide variety of disease states and pathologic processes. Mice deficient in IL-6 display abnormal and delayed liver regeneration and repair. Currently, IL-6 is thought to influence liver growth indirectly by priming hepatocytes to respond to growth factors such as hepatocyte growth factor (HGF) by inducing expression of HGF and by inhibiting hepatocyte apoptosis, as distinct from the direct mitotic effects of IL-6 on myeloid and other cell types. Here, we show that systemic administration of IL-6 using CHO cell tumors in nude mice results in dramatic hepatomegaly and hepatocyte hyperplasia in the absence of liver injury. Liver mass and liver to body mass ratios increased to 2 to 3 times normal because of proliferation of hepatocytes. Liver growth was associated with high levels of serum IL-6 and with activation of the IL-6-signaling pathway, including increased expression of IL-6 receptor-alpha/gp80, activation of the signal transducer and activator of transcription-3 (STAT-3), and mitogen-activated protein kinase (MAPK/ERK)-signaling pathways and induction of downstream target genes, including c-myc. HGF receptor and transforming growth factor alpha (TGF-alpha)/epidermal growth factor (EGF) receptor activation were decreased in hypertrophied livers, suggesting that IL-6-induced liver growth was independent of these known hepatocyte mitotic pathways. In conclusion, we suggest that IL-6 may function as a direct hepatic mitogen in vivo and, furthermore, that IL-6 warrants closer examination as a potent liver growth factor with potential clinical utility for increasing liver mass following injury. PMID- 12883477 TI - Nuclear factor kappa B decoy oligodeoxynucleotides prevent endotoxin-induced fatal liver failure in a murine model. AB - Endotoxin syndrome is a systemic inflammatory response mediated by inflammatory cytokines. Nuclear factor kappa B (NF-kappa B) is the dominant regulator of the production of these cytokines by inflammatory cells. The aim of this study was to assess the efficacy of in vivo transfer of synthetic double-stranded oligodeoxynucleotides (ODN) with high affinity against NF-kappa B (NF-kappa B/decoy/ODN) as a therapeutic strategy for treating endotoxin-induced fatal liver injury. Liver injury was induced by administration of lipopolysaccharide (LPS) to Propionibacterium acnes-primed BALB/C mice. NF-kappa B/decoy/ODN was transferred into the portal vein using a fusigenic liposome with hemagglutinating virus of Japan. NF-kappa B/decoy/ODN was preferentially transferred to Kupffer cells, and activation of NF-kappa B after the LPS challenge was suppressed, leading to decreased inflammatory cytokine production. As a result, the massive necrosis and hepatocyte apoptosis observed in the control mice was dramatically attenuated and the survival rate improved. In conclusion, NF-kappa B/decoy/ODN transfer in vivo effectively suppressed endotoxin-induced fatal liver injury in mice. PMID- 12883478 TI - Effects of proinflammatory cytokines on rat organic anion transporters during toxic liver injury and cholestasis. AB - Hepatobiliary transporters are down-regulated in toxic and cholestatic liver injury. Cytokines such as tumor necrosis factor alpha (TNF-alpha) and interleukin 1 beta (IL-1 beta) are attributed to mediate this regulation, but their particular contribution in vivo is still unknown. Thus, we studied the molecular mechanisms by which Ntcp, Oatp1, Oatp2, and Mrp2 are regulated by proinflammatory cytokines during liver injury. Rats were injected intraperitoneally with either carbon tetrachloride or endotoxin. Inactivation of TNF-alpha and IL-1 beta was achieved by repetitive intraperitoneal injection of etanercept and anakinra, respectively. Messenger RNA (mRNA) levels of transporters and binding activities as well as nuclear protein levels of Ntcp, Oatp2, and Mrp2 transactivators were determined 20 to 24 hours later. In contrast to IL-1 beta, TNF-alpha inactivation alone fully prevented down-regulation of Ntcp, Oatp1, and Oatp2 mRNA as well as reduced binding activity of hepatocyte nuclear factor 1 (HNF-1) in CCl(4)-induced toxic injury. In endotoxemia, down-regulation of Mrp2, and partially in case of Ntcp, could be prevented by IL-1 beta but not TNF-alpha blockade. However, inactivation of either cytokine led to preservation of HNF1 and partially of retinoid X receptor/retinoic acid receptor (RXR/RAR) binding activity. No effect of anticytokines was seen on pregnane X receptor (PXR) and constitutive androstane receptor (CAR) binding activity as well as nuclear protein mass. In conclusion, TNF-alpha represents the master cytokine responsible for HNF1 dependent down-regulation of Ntcp, Oatp1, and Oatp2 in CCl(4)-induced toxic liver injury. IL-1 beta predominates in a complex signaling network of Ntcp and Mrp2 regulation in cholestatic liver injury. In contrast to in vitro studies, HNF1 and RXR/RAR-independent mechanisms appear to be more important in regulation of Mrp2 and Ntcp gene expression in endotoxemia. PMID- 12883479 TI - Neutrophils aggravate acute liver injury during obstructive cholestasis in bile duct-ligated mice. AB - Obstruction of the common bile duct in a variety of clinical settings leads to cholestatic liver injury. An important aspect of this injury is hepatic inflammation, with neutrophils as the prominent cell type involved. However, the pathophysiologic role of the infiltrating neutrophils during cholestatic liver injury remains unclear. Therefore, we tested the hypothesis that neutrophils contribute to the overall pathophysiology by using bile duct-ligated (BDL) wild type animals and mice deficient in the beta(2) integrin CD18. In wild-type animals, neutrophils were activated systemically as indicated by the increased expression of Mac-1 (CD11b/CD18) and L-selectin shedding 3 days after BDL. Histologic evaluation (48 +/- 10% necrosis) and plasma transaminase levels showed severe liver injury. Compared with sham-operated controls (< 10 neutrophils per 20 high-power fields), large numbers of neutrophils were present in livers of BDL mice (425 +/- 64). About 60% of these neutrophils had extravasated into the parenchyma. In addition, a substantial number of extravasated neutrophils were found in the portal tract. In contrast, Mac-1 was not up-regulated and plasma transaminase activities and the area of necrosis (21 +/- 9%) were significantly reduced in CD18-deficient animals. These mice had overall 62% less neutrophils in the liver. In particular, extravasation from sinusoids and portal venules (PV) was reduced by 91% and 47%, respectively. Immunohistochemical staining for chlorotyrosine, a marker of neutrophil-derived oxidant stress, was observed in the parenchyma of BDL wild-type but not CD18-deficient mice. In conclusion, neutrophils aggravated acute cholestatic liver injury after BDL. This inflammatory injury involves CD18-dependent extravasation of neutrophils from sinusoids and reactive oxygen formation. PMID- 12883480 TI - Bilirubin rinse: A simple protectant against the rat liver graft injury mimicking heme oxygenase-1 preconditioning. AB - Heme oxygenase (HO)-1 preconditioning through genetic or pharmacologic interventions was shown experimentally to improve posttransplant outcome of liver grafts. However, its clinical application requires careful consideration because of the complexity and economic costs of the procedures. This study aimed to examine if graft preconditioning with HO-1 could be substituted by a simple treatment with heme-degrading products such as bilirubin. Rats were pretreated with or without hemin, an HO-1 inducer for preconditioning. Their livers were harvested as grafts in University of Wisconsin (UW) solution for 16 hours at 4 degrees C and followed by reperfusion ex vivo or by transplantation in vivo. The control grafts were also treated with a rinse buffer containing varied concentrations of unconjugated bilirubin with different time intervals. The HO-1 preconditioned grafts ex vivo exhibited a marked improvement of bile output and cell injury that was cancelled by blocking HO with zinc protoporphyrin-IX. The aggravation of the graft viability by the inhibitor was repressed by supplementation of bilirubin but not by that of carbon monoxide. Furthermore, a short-term rinse treatment with micromolar levels of bilirubin attenuated biliary dysfunction and cell injury of the grafts both ex vivo and in vivo even without HO-1 preconditioning. The protective effects of HO-1 preconditioning or bilirubin rinse appeared to involve its inhibitory effects on lipid peroxidation in hepatocytes. In conclusion, these results suggest that bilirubin rinse serves as a simple strategy to ameliorate hyperacute oxidative stress and hepatobiliary dysfunction of the transplanted grafts, mimicking effects of HO-1-mediated preconditioning. PMID- 12883481 TI - Cotransport of reduced glutathione with bile salts by MRP4 (ABCC4) localized to the basolateral hepatocyte membrane. AB - The liver is the major source of reduced glutathione (GSH) in blood plasma. The transport protein mediating the efflux of GSH across the basolateral membrane of human hepatocytes has not been identified so far. In this study we have localized the multidrug resistance protein 4 (MRP4; ABCC4) to the basolateral membrane of human, rat, and mouse hepatocytes and human hepatoma HepG2 cells. Recombinant human MRP4, expressed in V79 hamster fibroblasts and studied in membrane vesicles, mediated ATP-dependent cotransport of GSH or S-methyl-glutathione together with cholyltaurine, cholylglycine, or cholate. Several monoanionic bile salts and the quinoline derivative MK571 were potent inhibitors of this unidirectional transport. The K(m) values were 2.7 mmol/L for GSH and 1.2 mmol/L for the nonreducing S-methyl-glutathione in the presence of 5 micromol/L cholyltaurine, and 3.8 micromol/L for cholyltaurine in the presence of 5 mmol/L S methyl-glutathione. Transport of bile salts by MRP4 was negligible in the absence of ATP or without S-methyl-glutathione. These findings identify a novel pathway for the efflux of GSH across the basolateral hepatocyte membrane into blood where it may serve as an antioxidant and as a source of cysteine for other organs. Moreover, MRP4-mediated bile salt transport across the basolateral membrane may function as an overflow pathway during impaired bile salt secretion across the canalicular membrane into bile. In conclusion, MRP4 can mediate the efflux of GSH from hepatocytes into blood by cotransport with monoanionic bile salts. PMID- 12883482 TI - Ethanol metabolism results in a G2/M cell-cycle arrest in recombinant Hep G2 cells. AB - Previous studies using the Hep G2-based VA cells showed that ethanol metabolism resulted in both cytotoxicity and impaired DNA synthesis, causing reduced accumulation of cells in culture. To further characterize the ethanol oxidation mediated impairment of DNA synthesis we analyzed the cell-cycle progression of VA cells. These studies showed approximately a 6-fold increase in the percentage of cells in the G2/M phase of the cell cycle after 4 days of ethanol exposure. The G2/M transition requires activity of the cyclin-dependent kinase, Cdc2. Cdc2 is positively regulated by association with cyclin B1, and negatively regulated by phosphorylation of amino acids Thr14 and Tyr15. Immunoblot analysis revealed that ethanol metabolism had little affect on total Cdc2 content in these cells, but resulted in the accumulation of up to 20 times the amount of cyclin B1, indicating that cyclin B1 was available for formation of Cdc2/cyclin B1 complexes. Co-immunoprecipitation revealed that 6 times more Cdc2/cyclin B1 complexes were present in the ethanol-treated cells compared with the controls. Investigation of the phosphorylation state of Cdc2 revealed that ethanol oxidation increased the amount of the phosphorylated inactive form of Cdc2 by approximately 3-fold. Thus, the impairment in cell-cycle progression could not be explained by a lack of cyclin B1, or the ability of Cdc2 and cyclin B1 to associate, but instead resulted, at least in part, from impaired Cdc2 activity. In conclusion, ethanol oxidation by VA cells results in a G2/M cell-cycle arrest, mediated by accumulation of the phosphorylated inactive form of Cdc2. PMID- 12883483 TI - Antimycin A-induced defenestration in rat hepatic sinusoidal endothelial cells. AB - Liver sinusoidal endothelial cells (LSECs) possess fenestrae arranged in sieve plates. Hepatic endothelial fenestrae are open pores approximately 100 to 200 nm in diameter. Alterations in their number or diameter by hormones, xenobiotics, and diseases have important implications for hepatic microcirculation and function. Numerous reports of hepatotoxin-induced defenestration suggest that the cytoskeleton and the energy status of hepatic endothelial cells play a key role in the regulation of fenestrae. Therefore, we investigated the effect of antimycin A, an inhibitor of mitochondrial energy production, on the number of fenestrae in cultured LSECs using high-resolution microscopy and immunocytochemistry. Prolonged incubation (greater than 30 min) with antimycin A resulted in defenestrated cells and coincided with the appearance of F-actin dots, whereas the distribution of G-actin remained unchanged. Adenosine triphosphate (ATP) was depleted dramatically to less than 5% within 30 minutes within the LSECs. After treatment with antimycin A, unusual elevated fenestrated complexes were apparent, organized as a meshwork of anastomosing fenestrae at the center of and above the sieve plates. The position and appearance of these novel structures and their association with defenestration suggest that they are implicated in the process of defenestration. In conclusion, the results of experiments with antimycin A suggest that ATP is needed to maintain fenestrae and the underlying fenestrae-associated cytoskeleton rings that maintain fenestrae patency. Antimycin A-induced defenestration of LSECs is associated with the development of a structure in the sieve plate that appears to be intrinsically involved in defenestration. PMID- 12883484 TI - A liver tolerates a portal antigen by generating CD11c+ cells, which select Fas ligand+ Th2 cells via apoptosis. AB - Administration of an antigen (Ag) per oral route leads to apoptosis of Ag specific CD4(+) T cells and to development of Th2 cells expressing Fas ligand (FasL) in the liver. We determined whether presentation of an ingested Ag in the liver alone was enough to select these FasL(+)Th2 cells and explored how this selection was achieved in the liver. Ovalbumin (OVA) administered orally was colocalized with class II(+) cells in the periportal and parenchymal area of the liver. On coculture with naive OVA-specific CD4(+) T cells, hepatic CD11c(+) cells from mice fed OVA generated Ag-specific Th2 cells. This was achieved by apoptosis of CD4(+) T cells, decrease of interleukin 12 (IL-12) secretion, and increase of IL-18 secretion by the CD11c(+) cells. Addition of IL-12 to this coculture prevented apoptosis of the CD4(+) T cells, which was associated with up modulation of IL-2 receptor beta chain expression. Administration of IL-12 to mice fed OVA prevented apoptosis of OVA-specific CD4(+) T cells in the liver. Moreover, adoptive transfer of hepatic CD11c(+) cells from mice fed OVA together with OVA-specific CD4(+) T cells led to development of Th2 cells as well as apoptosis of the transferred CD4(+) T cells in the lymph nodes of the recipient mice on immunization with OVA. In conclusion, presentation of an ingested Ag by hepatic CD11c(+) cells selects Th2 cells resistant to apoptosis in the liver, which is mediated in part by down-regulation of IL-12 secretion by the former cells. PMID- 12883485 TI - Cytokines and NASH: a pilot study of the effects of lifestyle modification and vitamin E. AB - There are few data evaluating plasma and/or peripheral blood monocyte cytokine concentrations/production or attempts to manipulate proinflammatory cytokines in nonalcoholic steatohepatitis (NASH). A pilot project in a general clinical research center evaluated the effects of a step 1 American Heart Association diet plus aerobic exercise with or without 800 IU of vitamin E daily on cytokine profiles and liver enzyme levels in 16 patients with biopsy-proven NASH. Biochemical assessment of liver function, lipid profiles, and body mass index significantly improved during the first 6 weeks of therapy and remained stable during the following 6 weeks. Plasma hyaluronic acid (HA) concentrations decreased in parallel with weight loss. Plasma tumor necrosis factor (TNF) concentrations were significantly elevated in patients with NASH and similar to patients with stable alcoholic cirrhosis but not as elevated as in patients with acute alcoholic steatohepatitis (AH). Although plasma TNF, interleukin 8 (IL-8), and IL-6 concentrations were all significantly elevated compared with control values, only plasma IL-6 levels significantly decreased with therapy. Peripheral blood monocyte TNF, IL-8, and IL-6 production was significantly elevated in patients with NASH but did not significantly decrease. Independent effects of vitamin E were not observed in this small sample. In conclusion, patients with NASH have dysregulated cytokine metabolism similar to, but less pronounced than abnormalities documented in AH. Cytokine values generally did not decrease significantly with weight loss with or without vitamin E over the duration of the study. Lifestyle modifications (low-fat diet and exercise) were associated with improvement in liver enzymes, cholesterol, and plasma HA levels in patients with NASH, whereas the level of vitamin E supplementation used in this short-term pilot study provided no apparent added benefit. PMID- 12883486 TI - Long-term outcomes of cirrhosis in nonalcoholic steatohepatitis compared with hepatitis C. AB - Data on the long-term outcome of nonalcoholic steatohepatitis (NASH)-associated cirrhosis are few, and most reports describe cases of cryptogenic cirrhosis associated with risk factors for NASH but without histologic definition. In this prospective cohort study, we describe the long-term morbidity and mortality of 23 patients with NASH-associated cirrhosis defined by strict clinicopathologic criteria. Outcomes were compared with 46 age- and gender-matched patients with cirrhosis from chronic hepatitis C virus (HCV) infection: 23 untreated and 23 nonresponders to antiviral therapy. During follow-up (mean, 84 months; median, 60 months; range, 5-177 months), 9 of the 23 NASH-associated cirrhosis cases developed liver-related morbidity (8 ascites and/or encephalopathy, 1 variceal bleeding). The probability of complication-free survival was 83%, 77%, and 48% at 1, 3, and 10 years, respectively, and the cumulative probability of overall survival was 95%, 90%, and 84% at 1, 3, and 10 years, respectively. Five deaths were from liver failure, 1 from a non-liver-related cause. By multivariate analysis, bilirubin (P =.02) and platelet (P =.04) were independent predictors of complication-free survival; bilirubin (P =.05) was the only predictor for overall survival. After controlling for these factors, there was no difference in complication-free or overall survival between the NASH-cirrhosis cohort and either group of HCV-cirrhosis. However, 8 cases of liver cancer occurred in the HCV-cirrhosis groups compared with none among NASH cases. In conclusion, liver failure is the main cause of morbidity and mortality in NASH-associated cirrhosis. The prognosis is either similar or less severe than HCV-cirrhosis, except that HCC appears less common. PMID- 12883487 TI - CYP2E1 activity before and after weight loss in morbidly obese subjects with nonalcoholic fatty liver disease. AB - Previous studies suggest that hepatic cytochrome P450 2E1 (CYP2E1) activity is increased in individuals with chronic alcoholism, nonalcoholic steatohepatitis (NASH), and morbid obesity, and may contribute to liver disease. We studied 16 morbidly obese subjects with varying degrees of hepatic steatosis and 16 normal weight controls. Obese subjects were evaluated at baseline, 6 weeks, and 1 year after gastroplasty, a procedure that leads to weight loss. Hepatic CYP2E1 activity was assessed by determination of the clearance of chlorzoxazone (CLZ), an in vivo CYP2E1-selective probe. Liver biopsy tissue was obtained during surgery for histopathology. Both the total and unbound oral CLZ clearance (Cl(u)/F) was elevated approximately threefold in morbidly obese subjects compared with controls (P <.001). The Cl(u)/F was significantly higher among subjects with steatosis involving >50% of hepatocytes, compared with those with steatosis in < or =50% of hepatocytes (P =.02). At postoperative week 6 and year 1, the median body mass index (BMI) of subjects who underwent gastroplasty decreased by 11% and 33%, total oral CLZ clearance declined by 16% (P <.01) and 46% (P <.05), and Cl(u)/F decreased by 18% (P <.05) and 35% (P =.16), respectively. Moreover, those subjects with a year 1 BMI <30 kg/m(2) exhibited a median Cl(u)/F that was 63% lower (P =.02) than the respective clearance for all other subjects. In conclusion, hepatic CYP2E1 activity is up-regulated in morbidly obese subjects. A positive association between the degree of steatosis and CYP2E1 activity preoperatively and between the extent of obesity and CYP2E1 activity postoperatively, suggests that CYP2E1 induction is related to or caused by hepatic pathology that results from morbid obesity. PMID- 12883488 TI - Prevention of diet-induced fatty liver in experimental animals by the oral administration of a fatty acid bile acid conjugate (FABAC). AB - Fatty acid bile acid conjugates (FABACs) are a new family of synthetic molecules designed to solubilize biliary cholesterol. They were shown to prevent and dissolve cholesterol gallstones in inbred C57L/J mice fed a lithogenic, high-fat diet (HFD). In these mice, fatty liver was observed in the controls but not in the FABAC-treated ones. The present study was designed to study the effect of FABAC (arachidyl-amido-cholanoic acid) on diet-induced fatty liver in rats, hamsters, and mice. The fatty liver score (on a scale of 0-4 by light microscopy) was 4.0 in control hamsters and 0.3 in the FABAC-fed hamsters (P <.001). In mice it was 1.5 and 0.4, respectively (P <.01). The lipid/protein ratio in the liver was 1.3 +/- 0.44 (mg lipid/mg protein) in control rats and 0.66 +/- 0.04 in the FABAC group (P =.001) after 14 days. In hamsters it was 1.41 +/- 0.27 and 1.11 +/ 0.20, respectively (P =.03), after 21 days. In Imperial Charles River (ICR) mice the ratio was 0.34 +/- 0.10 and 0.17 +/- 0.07 (P =.03), respectively, after 24 days. Liver fat concentration, measured as mg lipid/g liver tissue, decreased similarly by FABAC feeding. The decrease in liver fat affected mainly the triglyceride levels. FABAC-fed animals gained weight similarly to the controls. Triglyceride absorption was unaffected by FABAC supplementation. In conclusion, oral FABAC therapy prevents/reduces the development of fatty liver in animals consuming a HFD. PMID- 12883489 TI - Fulminant hepatitis after grand mal seizures: mechanisms and role of liver transplantation. AB - Fulminant liver failure is a rare complication of grand mal seizures with a high mortality, the prognosis being largely determined by the combination of the hepatic and neurologic insults. The mechanisms of acute liver failure secondary to grand mal epilepsy and the place of liver transplantation in this context are poorly defined and are the subject of this report. A series of 6 such patients is presented. All had a history of chronic primary or post-traumatic epilepsy and presented with acute liver failure shortly after a grand mal fit. Detailed accounts of background, presentation, and management are given and integrated with blood, radiologic, and histologic investigations. Two of the 6 patients survived, 1 making a full recovery and the other with neurologic sequelae. Two patients underwent liver transplantation but died with severe neurologic sequelae despite improving liver function. The remaining 2 patients were considered too ill to undergo liver transplantation and died in multiple organ failure. Liver histology from needle biopsy and/or native liver explants identified lesions compatible with a combination of steatosis and necrosis. Factor V and transaminase levels may allow early identification of patients in whom liver function is likely to improve spontaneously. In conclusion, the mechanisms of liver failure occurring after grand mal seizures appear multifactorial, including hypoxia, steatosis, and drug-induced components. The neurological prognosis and overall survival of these patients remains poor. PMID- 12883490 TI - Increased carbon monoxide production in patients with cirrhosis with and without spontaneous bacterial peritonitis. AB - Carbon monoxide, a product of the heme-oxygenase (HO) pathway, is an important endogenous vasoactive substance. Production of CO has not been assessed in human cirrhosis. The aim of this study was to assess production of CO in patients with cirrhosis with and without spontaneous bacterial peritonitis (SBP). CO concentration in the exhaled air and blood carboxyhemoglobin (COHb) levels, as estimates of total HO activity, were determined in 16 healthy subjects, 32 noninfected cirrhotic patients (20 with ascites), and 19 patients with SBP, all nonsmokers. Noninfected cirrhotic patients had a CO concentration in the exhaled air and COHb levels significantly higher compared with values of healthy subjects (2.3 +/- 0.2 ppm vs. 0.7 +/- 0.1 ppm and 1.0% +/- 0.1% vs. 0.6% +/- 0.1%, respectively; P <.05 for both). Patients with ascites had the highest values. Both CO concentration in the exhaled air and COHb levels were very high in patients with SBP (5.6 +/- 0.6 ppm and 1.9% +/- 0.2%; P <.01 vs. the other 2 groups) and decreased slowly after resolution of the infection, reaching values similar to those of noninfected patients 1 month after SBP. In patients with SBP, there was a significantly direct correlation between CO and plasma renin activity (PRA) (r = 0.71, P <.001). In conclusion, these results support the existence of increased CO production in human cirrhosis, which further increases in the setting of SBP. Increased CO production may participate in the disturbance of circulatory function that occurs during severe bacterial infections in cirrhosis. PMID- 12883491 TI - Validation and refinement of survival models for liver retransplantation. AB - Orthotopic liver retransplantation (re-OLT) is highly controversial. The objectives of this study were to determine the validity of a recently developed United Network for Organ Sharing (UNOS) multivariate model using an independent cohort of patients undergoing re-OLT outside the United States, to determine whether incorporation of other variables that were incomplete in the UNOS registry would provide additional prognostic information, to develop new models combining data sets from both cohorts, and to evaluate the validity of the model for end-stage liver disease (MELD) in patients undergoing re-OLT. Two hundred eighty-one adult patients undergoing re-OLT (between 1986 and 1999) at 6 foreign transplant centers comprised the validation cohort. We found good agreement between actual survival and predicted survival in the validation cohort; 1-year patient survival rates in the low-, intermediate-, and high-risk groups (as assigned by the original UNOS model) were 72%, 68%, and 36%, respectively (P <.0001). In the patients for whom the international normalized ratio (INR) of prothrombin time was available, MELD correlated with outcome following re-OLT; the median MELD scores for patients surviving at least 90 days compared with those dying within 90 days were 20.75 versus 25.9, respectively (P =.004). Utilizing both patient cohorts (n = 979), a new model, based on recipient age, total serum bilirubin, creatinine, and interval to re-OLT, was constructed (whole model chi(2) = 105, P <.0001). Using the c-statistic with 30-day, 90-day, 1-year, and 3-year mortality as the end points, the area under the receiver operating characteristic (ROC) curves for 4 different models were compared. In conclusion, prospective validation and use of these models as adjuncts to clinical decision making in the management of patients being considered for re-OLT are warranted. PMID- 12883492 TI - Cellular retinol-binding protein-1 in hepatocellular carcinoma correlates with beta-catenin, Ki-67 index, and patient survival. AB - The cellular retinol-binding protein-1 (CRBP-1) plays a key role in the esterification and intercellular transfer of retinol. By in situ hybridization, immunohistochemistry, and confocal laser scanning microscopy (CLSM), we show that, in normal liver, CRBP-1 is strongly expressed in the cytoplasm of hepatic stellate cells (HSCs) and myofibroblasts (MFs) with only low CRBP-1 levels in hepatocytes. By contrast, in 196 hepatocellular carcinoma (HCC) specimens CRBP-1 expression in MFs was down-regulated in 83%. Patients with high CRBP-1 expression in MFs had a significantly higher 2-year survival as compared with patients with low CRBP-1 expression (52% vs. 29%, respectively; P =.034). An aberrant nuclear CRBP-1 accumulation resulting from cytoplasmic invagination was found in 29% of HCCs. Nuclear CRBP-1 staining correlated positively with a favorable tumor stage (Okuda stage I; P =.01) and negatively with the Ki-67(+) proliferation fraction (PF). A Ki-67(+) PF of > or =10% was associated with a lower 2-year survival probability as compared with patients with a Ki-67(+) PF of <10% (12% vs. 40%, respectively; P =.015). Prognosis did not correlate with the nuclear beta-catenin expression. There was, however, a close correlation between nuclear CRBP-1 inclusions and nuclear beta-catenin staining in HCCs (P =.008), suggesting a cross talk between CRBP-1 and the Wnt/wingless signal transduction pathway. In conclusion, our findings demonstrate that CRBP-1 detection may be useful for the discrimination between nonneoplastic and neoplastic liver cells and suggest that modulation of CRBP-1 expression in HCCs contributes to tumor growth and progression via retinoid-mediated signaling and disruption of cellular vitamin A homeostasis. PMID- 12883493 TI - Biochemical surrogate markers of liver fibrosis and activity in a randomized trial of peginterferon alfa-2b and ribavirin. AB - Liver fibrosis and activity indexes were validated in patients infected by hepatitis C virus (HCV) nontreated and treated by interferon. The aim was to validate their usefulness as surrogate markers of histologic features using the data of a randomized trial of combination peginterferon alfa-2b and ribavirin. Three hundred fifty-two patients who had had 2 interpretable liver biopsies and stored serum sample before and after treatment were selected. Two hundred eight patients received peginterferon alfa-2b 1.5 mcg per kg and ribavirin and 144 patients interferon alfa-2b 3 MU three times a week and ribavirin for 48 weeks. A fibrosis and an activity index combining 5 and 6 biochemical markers were assessed at baseline and at end of follow-up (24 weeks after treatment). The biochemical markers have significant predictive values both for the diagnosis of fibrosis and for activity. For the diagnosis of bridging fibrosis and/or moderate necroinflammatory activity, the area under the receiver operating characteristics curve of the activity index was 0.76 +/- 0.03 at baseline and 0.82 +/- 0.02 at end of follow-up. A cutoff of activity index at 0.30 (range, 0.00-1.00) had 90% sensitivity and 88% positive predictive value for the diagnosis of bridging fibrosis or moderate necroinflammatory activity. Sensitivity analyses with biopsy specimens of size greater than 15 mm suggest that a part of discordances between biochemical markers and histology were due to biopsy specimen sampling error. In conclusion, these biochemical markers of fibrosis and activity could be used as surrogate markers for liver biopsy in patients with chronic hepatitis C, both for the initial evaluation and for follow-up. PMID- 12883495 TI - A potent and specific morpholino antisense inhibitor of hepatitis C translation in mice. AB - Hepatitis C virus (HCV) is an RNA virus infecting one in every 40 people worldwide. Current treatments are ineffective and HCV is the leading cause of liver failure leading to transplantation in the United States and Europe. Translational control of HCV is a prime therapeutic target. We assessed the inhibitory potential of morpholino phosphoramidate antisense oligonucleotides (morpholinos) on HCV translation by codelivering them with reporter plasmids expressing firefly luciferase under the translational control of the HCV internal ribosome entry site (IRES) into the livers of mice. Real-time imaging of HCV IRES luciferase reporter messenger RNA (mRNA) translation in living mice showed that a 20-mer complementary to nucleotides 345-365 of the IRES inhibited translation by greater than 95% for at least 6 days and showed mismatch specificity. No significant nonspecific inhibition of a cap-dependent luciferase or encephalomyocarditis virus (EMCV) IRES luciferase reporter translation was observed. Inhibition by the 20-mer morpholino was dose dependent, with 1 nmol/mouse giving the highest inhibition. In conclusion, morpholino antisense oligonucleotides are potent inhibitors of HCV IRES translation in a preclinical mouse model; morpholinos have potential as molecular therapeutics for treating HCV and other viral infections. The in vivo model described is a broadly applicable, straightforward, and rapid readout for inhibitor efficacy. As such, it will greatly facilitate the development of novel therapeutic strategies for viral hepatitis. Notably, the level of antisense inhibition observed in this in vivo model is similar to the maximal inhibition we have obtained previously with RNA interference in mice. PMID- 12883494 TI - Favorable prognosis of chronic hepatitis C after interferon therapy by long-term cohort study. AB - The prognosis of patients with chronic hepatitis C after interferon (IFN) therapy is still poorly defined. The present study evaluated the effect of IFN therapy on survival in a cohort of such patients. The study included 459 patients with biopsy-proven C-viral chronic liver disease who were followed for 8.2 +/- 2.9 years (range, 7-183 months). Survival status was examined by medical records or direct questionnaires. Fifteen (14%) of 104 IFN-untreated patients and 33 (9%) of 355 patients treated with IFN died during follow-up. Among the treated patients, 4 (3%) of 116 with sustained virologic response and 29 (12%) of 239 without sustained virologic response died. Liver-related death was shown in 32 (67%) patients, and hepatocellular carcinoma (HCC) caused 25 (52%) of the 48 deaths. Multivariate Cox proportional regression analysis revealed that IFN treatment decreased the risk ratio for overall death to 0.521 (confidence interval [CI]: 0.263-1.034) and for liver-related death to 0.208 (CI: 0.088-0.495) compared with untreated patients, and sustained virologic response showed a decrease in the risk ratio for overall death to 0.219 (CI: 0.068-0.710) and for liver-related death to 0.030 (CI: 0.003-0.267). IFN treatment showed no association with liver unrelated death. Furthermore, the standardized mortality ratios for all causes of death and liver-related death were reduced in IFN-treated patients compared with untreated patients (1.4 vs. 2.0 for total death and 7.9 vs. 19.7 for liver related death). In conclusion, the present data suggest that IFN therapy has a long-term clinical benefit for patients with chronic hepatitis C patients by reducing liver-related death, especially in patients with sustained virologic response. PMID- 12883496 TI - Dynamics of alanine aminotransferase during hepatitis C virus treatment. AB - Studies of the kinetics of hepatitis C virus (HCV) decline during interferon (IFN)-based therapy have led to insights into treatment efficacy. However, the kinetics of serum alanine aminotransferase (ALT), an enzyme used as a surrogate of liver damage, have not been closely monitored, and it is not known if they correlate with those of HCV RNA. Here we describe the associations between ALT and HCV dynamics. We analyzed 35 patients treated daily with 10 mIU IFN-alpha 2b with or without ribavarin for 28 days followed by standard IFN/ribavirin therapy. Patients exhibited 4 patterns of ALT change: (1) exponential decay of ALT, (2) transient increase in ALT followed by a decrease to pretreatment or normal levels, (3) increase in ALT to a new level, and (4) no significant change. By simultaneously modeling HCV and ALT dynamics, we successfully fit the observed changes. We found ALT decays with t(1/2) = 12.7 hours. The transient increase in ALT observed in some patients suggested a mild hepatotoxic effect of IFN. However, patients with a smaller initial ALT increase achieved higher rates of viral negativity by week 72 (P =.02). The week-4 ALT decline correlated with the HCV log drop (P =.006) and the efficacy of therapy (P =.025). In conclusion, our results suggest the use of ALT as a surrogate marker for treatment effect in patients with elevated ALT. PMID- 12883497 TI - A simple noninvasive index can predict both significant fibrosis and cirrhosis in patients with chronic hepatitis C. AB - Information on the stage of liver fibrosis is essential in managing chronic hepatitis C (CHC) patients. However, most models for predicting liver fibrosis are complicated and separate formulas are needed to predict significant fibrosis and cirrhosis. The aim of our study was to construct one simple model consisting of routine laboratory data to predict both significant fibrosis and cirrhosis among patients with CHC. Consecutive treatment-naive CHC patients who underwent liver biopsy over a 25-month period were divided into 2 sequential cohorts: training set (n = 192) and validation set (n = 78). The best model for predicting both significant fibrosis (Ishak score > or = 3) and cirrhosis in the training set included platelets, aspartate aminotransferase (AST), and alkaline phosphatase with an area under ROC curves (AUC) of 0.82 and 0.92, respectively. A novel index, AST to platelet ratio index (APRI), was developed to amplify the opposing effects of liver fibrosis on AST and platelet count. The AUC of APRI for predicting significant fibrosis and cirrhosis were 0.80 and 0.89, respectively, in the training set. Using optimized cut-off values, significant fibrosis could be predicted accurately in 51% and cirrhosis in 81% of patients. The AUC of APRI for predicting significant fibrosis and cirrhosis in the validation set were 0.88 and 0.94, respectively. In conclusion, our study showed that a simple index using readily available laboratory results can identify CHC patients with significant fibrosis and cirrhosis with a high degree of accuracy. Application of this index may decrease the need for staging liver biopsy specimens among CHC patients. PMID- 12883498 TI - Cytokine network during NASH and possible treatment selection. PMID- 12883499 TI - Drug-induced hepatic injury in Sweden. PMID- 12883500 TI - Absence of Helicobacter species in the liver of patients with primary or metastatic liver cancer. PMID- 12883501 TI - More on serum phosphate and prognosis of acute liver failure. PMID- 12883502 TI - Cost effectiveness of preventing variceal hemorrhage. PMID- 12883503 TI - Emergency department crowding: a point in time. AB - STUDY OBJECTIVE: This is a pilot study designed to assess the feasibility of a point prevalence study to assess the degree of crowding in hospital emergency departments (EDs). In addition, we sought to measure the degree of physical crowding and personnel shortage in our sample. METHODS: A mail survey was sent to a random sample of 250 EDs chosen from a database compiled by the American College of Emergency Physicians of 5,064 EDs in the United States. In addition to demographic information, respondents were asked to count the patients and staff in their EDs at 7 PM local time on Monday, March 12, 2001 (index time). RESULTS: The response rate was 36%. At the index time, there was an average of 1.1 patients per treatment space, and 52% of EDs reported more than 1 patient per treatment space. There was also evidence of personnel shortage, with a mean of 4.2 patients per registered nurse and 49% of EDs having each registered nurse caring for more than 4 patients. There was a mean of 9.7 patients per physician. Sixty-eight percent of EDs had each physician caring for more than 6 patients. There was crowding present in all geographic areas and all hospital types (teaching-nonteaching status of the hospital). Consistent with the crowded conditions, 11% of institutions were on ambulance diversion and not accepting new acute patients. Delays in transfer of admitted patients out of the ED contributed to the physical crowding. Twenty-two percent of patients in the ED were already admitted and were awaiting transfer to an inpatient bed; 73% of EDs were boarding 2 or more inpatients. The amount of crowding quantified by this point prevalence study was confirmed by the amount of crowding reported for the previous week: 48% of EDs were boarding inpatients during the previous week for a mean of 8.9 hours, 4.2 days per week; 31% had been on diversion; 59% had been routinely using their halls for patients; 38% had been doubling their rooms; and 47% had been using nonclinical space for patient care. CONCLUSION: Our low response rate limits this pilot study. Nonetheless, this study, as well as others, demonstrates that EDs throughout the United States are severely crowded. Such crowding raises concerns about the ability of EDs to respond to mass casualty or volume surges. PMID- 12883504 TI - A conceptual model of emergency department crowding. AB - Emergency department (ED) crowding has become a major barrier to receiving timely emergency care in the United States. Despite widespread recognition of the problem, the research and policy agendas needed to understand and address ED crowding are just beginning to unfold. We present a conceptual model of ED crowding to help researchers, administrators, and policymakers understand its causes and develop potential solutions. The conceptual model partitions ED crowding into 3 interdependent components: input, throughput, and output. These components exist within an acute care system that is characterized by the delivery of unscheduled care. The goal of the conceptual model is to provide a practical framework on which an organized research, policy, and operations management agenda can be based to alleviate ED crowding. PMID- 12883505 TI - Calculus, chaos, and other models of emergency department crowding. PMID- 12883506 TI - Specificity and sensitivity of automated external defibrillator rhythm analysis in infants and children. AB - STUDY OBJECTIVES: The rhythm detection algorithms of automated external defibrillators have been derived from adult rhythms, and their ability to discriminate between shockable and nonshockable rhythms in children is largely unknown. This study evaluates the performance of 1 automated external defibrillator algorithm in infants and children and evaluates algorithm performance with anterior-posterior versus sternal-apex lead placement. METHODS: We enrolled pediatric patients in a critical care unit, an electrophysiology laboratory, and a cardiac operating room. A monitor-defibrillator recorded ECGs by means of standard defibrillation-monitor pads. Selected 15-second rhythm samples were played into a LIFEPAK 500 automated external defibrillator, and the automated external defibrillator "shock/no shock" decision was documented. To determine sensitivity and specificity, the automated external defibrillator decision was compared with the "shockable" versus "nonshockable" rhythm classification provided by 3 expert clinicians who were blinded to the automated external defibrillator decision. RESULTS: We recorded 1,561 rhythm samples from 203 pediatric patients (median age 11 months; range, day of birth to 7 years). The automated external defibrillator recommended a shock for 72 of 73 rhythm samples classified as coarse ventricular fibrillation by expert review (sensitivity 99%; 95% confidence interval [CI] 93% to 100%); and correctly reached a "no shock advised" decision for 1,465 of 1,472 rhythm samples classified as nonshockable by experts (specificity 99.5%). Specificity was 99.1% (95% CI 97.8% to 99.8%) with the sternal-apex lead and 99.4% (95% CI 98.1% to 99.9%) with the anterior-posterior lead. CONCLUSION: This automated external defibrillator algorithm has high specificity and sensitivity when used in infants and children with either sternal-apex or anterior-posterior lead placement. PMID- 12883507 TI - Emergency department analgesia for fracture pain. AB - STUDY OBJECTIVES: We analyze records of all emergency department (ED) patients with extremity or clavicular fractures to describe analgesic use, compare analgesia between adults and children, and compare analgesia between the subset of these adults and children with documented moderate or severe pain. Among children, we compare treatment between pediatric and nonpediatric facilities. METHODS: Analysis of the ED component of the National Center for Health Statistics National Hospital Ambulatory Medical Care Survey for 1997 through 2000 was conducted. The proportion of patients with closed extremity and clavicular fracture that received any analgesic and narcotic analgesic medications was determined for each age category. Survey-adjusted regression analyses compared pain and narcotic medications by age and ED type (pediatric versus other). Analyses were repeated for the subset of patients with moderate or severe pain severity scores. RESULTS: Of 2,828 patients with isolated closed fractures of the extremities or clavicle, 64% received any analgesic and 42% received a narcotic analgesic. Pain severity scores were recorded for 59% of visits overall, 47% of children younger than 4 years, and 34% of children younger than 1 year. Among patients with documented moderate or severe pain, 73% received an analgesic and 54% received a narcotic analgesic. Compared with adults, a lower proportion of children (< or = 15 years) received either any analgesic or a narcotic analgesic (P <.001). After adjustment for confounders and survey design, the proportion of patients aged 0 to 3, 4 to 8, 9 to 15, 16 to 29, 30 to 69, and 70 years and older who received any analgesic was 54% (95% confidence interval [CI] 41% to 67%), 63% (95% CI 57% to 68%), 60% (95% CI 57% to 64%), 67% (95% CI 62% to 73%), 68% (95% CI 64% to 72%), and 58% (95% CI 52% to 65%), respectively; the proportion who received a narcotic analgesic was 21% (95% CI 11% to 31%), 30% (95% CI 22% to 37%), 27% (95% CI 23% to 32%), 47% (95% CI 40% to 54%), 51% (95% CI 46% to 56%), and 41% (95% CI 35% to 48%), respectively. Compared with children treated in other EDs, children treated in pediatric EDs were about as likely to receive any analgesia (adjusted relative risk [RR] 1.1; 95% CI 0.9 to 1.3) or narcotic analgesia (adjusted RR 0.9; 95% CI 0.6 to 1.2). CONCLUSION: In pediatric and adult patients, pain medications were frequently not part of ED treatment for fractures, even for visits with documented moderate or severe pain. Pain severity scores were often not recorded. Pediatric patients were least likely to receive analgesics, especially narcotics. PMID- 12883508 TI - The role of the emergency physician in emergency medical services for children. AB - Emergency physicians have a duty to advance the care of pediatric patients in the emergency medical services (EMS) system. This policy resource and education paper, designed to support the American College of Emergency Physicians policy paper "The Role of the Emergency Physician in Emergency Medical Services for Children," describes the development of the federal EMS for Children Program, the importance of the integration of EMS for children into EMS systems, and the role of the emergency physician in EMS for children. PMID- 12883509 TI - Identifying febrile young infants with bacteremia: is the peripheral white blood cell count an accurate screen? AB - STUDY OBJECTIVE: We estimated the accuracy of the total peripheral WBC count as a screen for bacteremia in febrile young infants. METHODS: We evaluated, retrospectively, the performance characteristics of linear and nonlinear (U shaped) logistic models for predicting bacteremia that are based on the total peripheral WBC count. Research subjects were consecutive 0- to 89-day-old infants who had a temperature in triage of greater than or equal to 38 degrees C (> or =100.4 degrees F) and were evaluated for infection at a pediatric emergency department (1993 to 1999). Infants with leukemia were excluded. Areas under the receiver operator characteristic curves (AUC), as well as sensitivity, specificity, interval likelihood ratios, and the corresponding odds of bacteremia predicted at various thresholds of the test, were calculated. RESULTS: The rate of bacteremia was 1% (38/3,810). The U-shaped model was more accurate (AUC 0.69 versus 0.56); however, no threshold of the total peripheral WBC count had both good sensitivity and specificity. Sensitivity and specificity values were 79% and 5%, respectively, at a peripheral WBC count cutoff of 5,000 cells/mm(3), and 45% and 78%, respectively, at a cutoff of 15,000 cells/mm(3). The odds of bacteremia were not decreased substantially at any cutoff and were increased only modestly at values outside published norms of the test. CONCLUSION: The total peripheral WBC count is an inaccurate screen for bacteremia in febrile young infants; thus, decisions to obtain blood cultures should not rely on this test. PMID- 12883510 TI - Foodborne botulism in a six-month-old infant caused by home-canned baby food. AB - Previously reported cases of botulism in infants have been diagnosed as infant botulism; that is, botulism caused by intestinal colonization by Clostridium botulinum with intraluminal production and absorption of toxin. Foodborne botulism is caused by ingestion of preformed toxin. We describe an unusual case of foodborne botulism in a 6-month-old infant caused by the ingestion of improperly prepared home-canned baby food. This represents the youngest age of onset for foodborne botulism in the United States of which we are aware and illustrates the need to rule out foodborne botulism, which represents a public health emergency, regardless of the patient's age. The diagnosis could have been readily missed or delayed in this case because the weakness was rapidly progressive rather than insidious, as is typical of infant botulism. PMID- 12883511 TI - Dynamic nature of electrocardiographic waveform predicts rescue shock outcome in porcine ventricular fibrillation. AB - STUDY OBJECTIVE: Survival decreases with duration of ventricular fibrillation, and it is possible that failed rescue shocks increase myocardial damage. Structure in the ECG signal during ventricular fibrillation can be quantified by using the scaling exponent, a dimensionless measure that correlates with ventricular fibrillation duration. This study examined whether the scaling exponent could predict rescue shock success and whether unsuccessful rescue shocks altered the structure of the ventricular fibrillation waveform and the responsiveness to subsequent rescue shocks. METHODS: Ventricular fibrillation was electrically induced in 44 anesthetized swine, which were randomly assigned to receive 70-J biphasic rescue shocks at 2, 4, 6, 8, or 10 minutes. If rescue shocks failed, up to 2 subsequent rescue shocks were performed at 2-minute intervals. The scaling exponent was calculated at 1-second intervals from ECG to quantify the organization of the ventricular fibrillation waveform. RESULTS: A total of 92 rescue shocks were delivered, of which 23 successfully converted ventricular fibrillation to an organized rhythm (immediate success). After these 23 rescue shocks, 14 swine sustained organized rhythms for more than 30 seconds (sustained success). Lower scaling exponent values were associated with increased probability of successful rescue shocks. Receiver operating characteristic curves had an area under the curve of 0.86 for immediate rescue shock success and 0.93 for sustained rescue shock success. Failed rescue shocks increased the rate of scaling exponent increase over time but did not appear to affect subsequent rescue shock success when the scaling exponent was taken into account. CONCLUSION: Highly deterministic ventricular fibrillation, reflected by a low scaling exponent, predicted rescue shock success regardless of antecedent failed rescue shocks. In addition, unsuccessful rescue shocks might decrease post-rescue shock ventricular fibrillation waveform organization. PMID- 12883512 TI - Optimal defibrillation response intervals for maximum out-of-hospital cardiac arrest survival rates. AB - STUDY OBJECTIVE: Many centers optimize their emergency medical services (EMS) systems to achieve a target defibrillation response interval of "call received by dispatch" to "arrival at scene by responder with defibrillator" in 8 minutes or less for at least 90% of cardiac arrest cases. The objective of this study was to analyze survival as a function of time to test the evidence for this standard. METHODS: This prospective cohort study included all adult, cardiac etiology, out of-hospital cardiac arrest cases from phases I and II of the Ontario Prehospital Advanced Life Support (OPALS) study. Patients in the 21 Ontario study communities received a basic life support level of care with defibrillation by ambulance and firefighters but no advanced life support. Survival was plotted as a function of the defibrillation response interval. The equation of the curve, generated by means of logistic regression, was used to estimate survival at various defibrillation response interval cutoff points. RESULTS: From January 1, 1991, to December 31, 1997, there were 392 (4.2%) survivors overall among the 9,273 patients treated. The defibrillation response interval mean was 6.2 minutes, and the 90th percentile was 9.3 minutes. There was a steep decrease in the first 5 minutes of the survival curve, beyond which the slope gradually leveled off. Controlling for known covariates, the decrement in the odds of survival with increasing response interval was 0.77 per minute (95% confidence interval 0.74 to 0.83). The survival function predicts, for successive 90th percentile cutoff points, both survival rates and additional lives saved per year in the OPALS communities compared with the 8-minute standard: 9 minutes (4.6%; -18 lives), 8 minutes (5.9%; 0 lives), 7 minutes (7.5%; 23 lives), 6 minutes (9.5%; 51 lives), and 5 minutes (12.0%; 86 lives). CONCLUSION: The 8-minute target established in many communities is not supported by our data as the optimal EMS defibrillation response interval for cardiac arrest. EMS system leaders should consider the effect of decreasing the 90th percentile defibrillation response interval to less than 8 minutes. PMID- 12883513 TI - The eight-minute defibrillation response interval debunked: or is it? PMID- 12883514 TI - Public education regarding resuscitation: effects of a multimedia intervention. AB - STUDY OBJECTIVE: Previous studies have demonstrated inaccuracies in knowledge and perceptions regarding cardiopulmonary resuscitation (CPR) among the general public. This study was undertaken to determine the effect of a multimedia educational intervention on knowledge base and resuscitation preferences among the lay public. METHODS: In this prospective interventional study with preintervention and postintervention measurements, a validated multisite survey was administered to 310 volunteer lay participants in community-based settings during 2001 and 2002. The survey was piloted and validated (percentage of agreement index 98.6%; 95% confidence interval [CI] 0.9810 to 0.9900). An original 8-minute multimedia educational video was written and produced by physicians to provide educational information about cardiac resuscitation to the lay public. RESULTS: Among 310 participants, the mean age was 40 years (range 17 to 92 years), 67% were female, and 57% reported household incomes of more than 30,000 US dollars. Participants' median estimates of predicted postcardiac arrest survival rate before and after the educational intervention were 50% and 16%, respectively (median change 30%; 95% CI 25% to 35%). Median estimated durations of resuscitative efforts in the emergency department before and after the educational intervention were 30 minutes and 19 minutes, respectively (median change 10 minutes; 95% CI 5 to 15 minutes). For a series of hypothetical scenarios, significantly more participants indicated that they would refuse resuscitative efforts in scenarios involving terminally ill patients after the educational intervention. CONCLUSION: Inaccurate perceptions regarding cardiac resuscitation and postarrest survival exist among the lay public. A novel educational intervention demonstrated effective improvements in knowledge base regarding resuscitation, resulting in significant effects on resuscitation preferences among the lay public. Improved public education regarding resuscitation is needed to improve knowledge regarding CPR among the lay public. PMID- 12883515 TI - Osteopathic physicians in emergency medicine. PMID- 12883516 TI - Methodology for a rapid protocol to rule out pulmonary embolism in the emergency department. AB - We propose an emergency department (ED) pulmonary embolism rule-out protocol based on pretest probability assessment coupled with either a negative D -dimer assay result or a negative D -dimer assay result plus a normal alveolar dead space measurement. We examine the safety, efficiency, and feasibility of such a protocol, paying special attention to implicit and explicit strategies of pretest probability assessment among patients with suspected pulmonary embolism. Finally, we assess the potential effect of the proposed pulmonary embolism rule-out protocol on use of imaging resources and ED throughput. PMID- 12883517 TI - Feasibility of using an interactive laptop program in the emergency department to prevent alcohol misuse among adolescents. AB - STUDY OBJECTIVE: Alcohol, the most commonly used substance among adolescents, is frequently associated with injury. Effective interventions to prevent adolescent alcohol use and misuse in acute care settings are lacking. A laptop-based alcohol prevention program could reinforce other prevention efforts that adolescents may receive. We determined the feasibility of using an interactive laptop program with adolescent emergency department (ED) patients to prevent alcohol use and misuse. METHODS: We used the recruitment phase of a randomized controlled trial at an academic medical center and an urban teaching hospital. Patients were aged 14 to 18 years and presented within 24 hours of an acute injury. Measures included patient recruitment, mechanism of injury, injury severity score, alcohol use characteristics, and patients' opinion of the computer program. RESULTS: Of 843 eligible patients, 671 (79.6%) were enrolled and 655 (77.7%) completed the program. Parent or guardian reluctance was the most frequent reason for refusal. The participants averaged 16.0 years of age (range 14 to 18 years; SD 1.5 years), 66.9% were male, and 68.3% were white. Approximately 71% reported "ever" drinking. Recent alcohol use (past 3 months) by those "ever" drinking was as follows: 62.3% drank, 31.2% got drunk, and 37.4% binge drank. Seventy-four percent of recent drinkers reported that the program made them rethink their alcohol use. Ninety-four percent of participants liked the program. Only 5.3% required assistance with the program. CONCLUSION: Use of an interactive computer program in the ED appears feasible. Further work is being done to evaluate the effectiveness of the program in reducing alcohol-related behaviors among adolescents. PMID- 12883518 TI - Air bag-related injuries. PMID- 12883519 TI - Commentary: air bags: looking ahead through the rearview mirror. PMID- 12883520 TI - Pyrimidifen intoxication. AB - We present a case of lethal ingestion of pyrimidifen, a new insecticide with an unclear mode of action. The primary manifestations were coma and circulatory shock, leading to irreversible multiorgan failure. Pyrimidifen was detected in the patient's blood, urine, brain tissue, and gastric content samples. Minimal structural homology exists between pyrimidifen and organochlorines. Currently, no antidote is available, and therapy is primarily supportive. PMID- 12883521 TI - Evidence-based emergency medicine/skills for evidence-based emergency care. Interval likelihood ratios: another advantage for the evidence-based diagnostician. AB - Emergency physicians are often confronted with making diagnostic decisions on the basis of a test result represented on a continuous scale. When the results of continuous data are expressed as binary outcomes using a single cutoff, loss of information and distortion may occur. In this setting, interval likelihood ratios provide a distinct advantage in interpretation over those based on a dichotomized sensitivity and specificity. Dividing the data into intervals uses more of the information contained in the data and allows the clinician to more appropriately interpret the test results and to make valid clinical decisions. This article illustrates the advantages of interval likelihood ratios with examples and demonstrates how to calculate them on the basis of different data formats. Authors and journals need to be encouraged to report the results of studies of performance of diagnostic tests using interval ranges rather than simple dichotomization when the tests involve continuous variables. PMID- 12883522 TI - Evidence-based emergency medicine/editorial. The problem with sensitivity and specificity... PMID- 12883523 TI - Gene therapy progress and prospects: gene therapy of lysosomal storage disorders. AB - Despite disappointments with early clinical studies, there is continued interest in the development of gene therapy for the group of metabolic diseases referred to as lysosomal storage disorders (LSDs). The LSDs are monogenic and several small and large, representative animal models of the human diseases are available. Further, the successful reconstitution of only low and unregulated tissue levels of the affected lysosomal enzymes are expected to be sufficient to correct the disease at least in the case of some of the LSDs. For these reasons, they are perceived as good models for the evaluation of different gene delivery vectors and of different strategies for treating chronic genetic diseases by gene transfer. In this review, we will highlight the progress that has been made over the past 2 years in preclinical research for this group of disorders and speculate on future prospects. PMID- 12883524 TI - Poly (D, L-lactide-co-glycolide)/DNA microspheres to facilitate prolonged transgene expression in airway epithelium in vitro, ex vivo and in vivo. AB - Repeat administration of gene therapy for cystic fibrosis is likely to be essential for long-term clinical efficacy. This may be minimized by the use of slow-release gene transfer preparations with more prolonged expression and longer dosing intervals for the patient. Poly(D-L-lactide-co-glycolide) (PLG) is a biodegradable and biocompatible polymer that has been used to encapsulate plasmid DNA. PLG-DNA microspheres were generated and characterized with respect to morphology, size (80% of particles <5.2 microm), and encapsulation efficiency (50.7+/-2.3%, n=6). Gel electrophoresis of DNA re-extracted from the microspheres confirmed that despite a decrease in the proportion of supercoiled conformation, it had not been degraded by the preparation process. Gene transfer efficiency was tested using microspheres encapsulating the reporter gene beta-galactosidase in vitro on Cos 7 cells and a CF airway epithelial line (CFTEo approximately ) and ex vivo in a sheep tracheal (s.t.) model. In both cases, transgene expression was significantly (P<0.01) lower at the first time point tested (24 h in vitro, 48 h ex vivo) compared to lipid-#67-mediated gene transfer. However, PLG-mediated expression in vitro was sustained at 48 h, while lipid #67-mediated expression levels had dropped significantly (P<0.05) to 50.3+/-13.7 and 38.2+/-2.7% (Cos 7 and CFTEo approximately cells, respectively) of the 24-h level. This pattern was also seen in the s.t. model where at 72 h, PLG-mediated expression was 125.4+/ 7.2% of the 48-h level demonstrating significantly (P<0.05) better retention of transfection efficiency than lipid #67, where levels had fallen to approximately half the 48 h level. By 96 h, expression was still retained in the PLG transfected group (87.3+/-12.5% of 48 h expression) but was undetectable in the lipid -#67-transfected s.t. Finally, PLG microspheres, encapsulating the reporter gene chloramphenicol transferase (CAT, 80 microg) were instilled intranasally into Balb/C mice. Compared to lipid-#67-mediated delivery, where whole lung CAT expression was highest at 48 h (13.7 x 10(3)+/-0.05 CAT U/microg protein, n=6) and then not detectable at further time points, CAT expression was not detectable in PLG-transfected mice at 48 h, but was detectable at 7, 14 and 21 days after transfection. These data demonstrate that PLG-mediated gene transfer can produce prolonged gene expression in airway epithelia. However, gene transfer efficiency still requires significant improvement. PMID- 12883525 TI - Osteoinduction by ex vivo adenovirus-mediated BMP2 delivery is independent of cell type. AB - The objective of the study was to analyze and compare the abilities of various human cell types with inherently dissimilar osteogenic potentials to induce heterotopic bone formation following ex vivo transduction with two distinct adenoviral vectors encoding bone morphogenetic protein type 2 (BMP2). The cells comprised primary human bone marrow mesenchymal stem cells (BM-MSCs), primary human skin fibroblasts (SFs), and a human diploid fetal lung cell line (MRC-5). The vectors included adenovirus type 5 or a chimeric adenovirus type 5 with the fiber gene of adenovirus type 35 (Ad5F35-BMP2), both demonstrating significantly different expression of BMP2 in vitro. The experimental groups consisted of the three human cell types transduced with each of the two adenoviral vectors. Using nonobese diabetic severe combined immunodeficiency (NOD/SCID) mice, the transduced cells were injected intramuscularly following ex vivo adenoviral transduction. The nature and extent of heterotopic bone formation were analyzed radiographically and histologically. At 14 days postinjection, abundant, highly mineralized bone was formed in mice injected with Ad5F35-BMP2-transduced cells irrespective of the cell type. There was no statistically significant difference in the amount of bone formed between BM-MSCs, SFs, and MRC-5 cells transduced with Ad5F35-BMP2, as assessed from bone surface area on biplanar plain radiography. Substantially lesser amounts or no bone could be detected in mice injected with cells transduced with Ad5-BMP2. Immunohistochemical analysis confirmed the presence of human cells in muscle as early as 2 days postdelivery; however, at 6-7 days after injection, the transduced cells could not be detected in surrounding muscle, or in the heterotopic bone, indicating the host origin of the newly formed bone. The results of the study demonstrate no significant difference in osteoinductive properties between BM-MSCs, SFs, and MRC-5 cells transduced ex vivo with the same type of adenovirus encoding BMP2. The level of BMP2 expression appears to be a crucial factor determining the extent of heterotopic bone formation and was significantly affected by the type of adenovirus used. In the cell types studied, Ad5F35-BMP2 was more efficacious than Ad5-BMP2 in providing adequate levels of BMP2 for efficient osteoinduction. PMID- 12883526 TI - Hydrodynamics-based delivery of the viral interleukin-10 gene suppresses experimental crescentic glomerulonephritis in Wistar-Kyoto rats. AB - Gene therapy is expected to revolutionize the treatment of kidney diseases. Viral interleukin (vIL)-10 has a variety of immunomodulatory properties. We examined the applicability of vIL-10 gene transfer to the treatment of rats with crescentic glomerulonephritis, a T helper 1 (Th 1) predominant disease. To produce the disease, Wistar-Kyoto rats were injected with a rabbit polyclonal anti-rat glomerular basement membrane antibody. After 3 h, a large volume of plasmid DNA expressing vIL-10 (pCAGGS-vIL-10) solution was rapidly injected into the tail vein. pCAGGS solution was similarly injected into control rats (pCAGGS rats). We confirmed the presence of vector-derived vIL-10 mainly in the liver and observed high serum vIL-10 levels in pCAGGS-vIL-10-injected rats. Compared with the pCAGGS rats, the pCAGGS-vIL-10 rats showed significant therapeutic effects: reduced frequency of crescent formation, decrease in the number of total cells, macrophages, and CD4+ T cells in the glomeruli, decrease in urine protein, and attenuation of kidney dysfunction. Using quantitative real-time polymerase chain reaction, we also observed that this model was Th1-predominant in the glomeruli and that the ratio of the transcripts of CD4, interferon-gamma, tumor necrosis factor-alpha, and monocyte chemotactic protein-1 to the transcripts of glucose-6 phosphate dehydrogenase in the glomeruli were all significantly lower in the pCAGGS-vIL-10 rats than in the pCAGGS rats. These results demonstrate that pCAGGS vIL-10 gene transfer by hydrodynamics-based transfection suppresses crescentic glomerulonephritis. PMID- 12883527 TI - CD8+ T cells, NK cells and IFN-gamma are important for control of tumor with downregulated MHC class I expression by DNA vaccination. AB - One of the major hurdles facing cancer immunotherapy is that cancers may downregulate expression of MHC class I molecules. The development of a suitable tumor model with downregulated MHC class I expression is critical for designing vaccines and immunotherapeutic strategies to control such tumors. We developed an E7-expressing murine tumor model with downregulated MHC class I expression, TC-1 P3 (A15). Using this model, we tested DNA and vaccinia vaccines for their ability to control tumors with downregulated MHC class I expression. We found that vaccination with DNA encoding E7 linked to Mycobacterial heat shock protein 70 (HSP70) generated a significant antitumor effect against TC-1 P3 (A15), while vaccination with E7/HSP70 vaccinia did not generate an appreciable antitumor effect. Lymphocyte depletion experiments revealed that both CD8+ T cells and NK cells were essential for the antitumor effect generated by E7/HSP70 DNA against TC-1 P3 (A15). Furthermore, tumor protection experiments using IFN-gamma knockout mice revealed that IFN-gamma was essential for the antitumor effect generated by E7/HSP70 DNA against TC-1 P3 (A15). Our results demonstrate that vaccination with E7/HSP70 DNA results in a significant antitumor effect against a neoplasm with downregulated MHC class I expression and the importance of CD8+ T cells, NK cells, and IFN-gamma in generating this antitumor effect. PMID- 12883528 TI - HSV-1 amplicon vector-mediated expression of ATM cDNA and correction of the ataxia-telangiectasia cellular phenotype. AB - Ataxia-telangiectasia (A-T) is an autosomal recessive disorder characterized by neurodegeneration, immunodeficiency, cancer predisposition, genome instability, and radiation sensitivity. Previous research has shown that it is possible to correct the hereditary deficiency A-T by DNA transfection in cell culture, but the large size of the ATM cDNA (9 kb) limits the use of many vector types for gene replacement. HSV-1 amplicon vectors provide a means to deliver large genes to cells efficiently and without toxicity. In this study, the FLAG-tagged cDNA for human ATM was inserted into an HSV-1 amplicon under control of the CMV promoter (designated as HGC-ATM). FLAG-ATM expression was confirmed in 293T/17 cells and human A-T fibroblasts (GM9607) after transduction, by immunoprecipitation, Western analysis, and immunocytochemistry. Functional recovery was assessed by two independent assays. First, in vitro kinase assay showed that vector-derived ATM in GM9607 cells could successfully phosphorylate wt p53 using recombinant GST-p53(1-101). Second, in A-T cells infected with the HGC-ATM vector, the extent of accumulation in G2/M phase at 24 h postirradiation was similar to that observed in cells with wild-type endogenous ATM and lower than that observed in A-T cells infected with a control vector. Thus, these vectors provide a tool to test the feasibility of HSV-amplicons as gene therapy vectors for A-T. PMID- 12883529 TI - Purging of leukemia-contaminated bone marrow grafts using suicide adenoviral vectors: an in vivo murine experimental model. AB - Autologous bone marrow transplantation is an alternative therapeutic option for acute myeloid leukemia patients lacking a compatible donor. However, bone marrow from these patients may contain residual leukemic cells that should be ideally eliminated prior to the infusion of the graft. With the aim of developing more efficient protocols of graft purging, adenoviral-mediated gene transfer protocols have been conducted. We studied whether suicide adenoviral vectors expressing the cytosine deaminase gene (AdCD) could be used for selectively killing leukemic WEHI-3B cells. The AdCD transduction followed by the 5-fluorocytosine exposure abrogated the growth of WEHI-3B cells in vitro, with a minimal effect on normal hematopietic progenitors. To test the efficacy of the purging protocol in vivo, bone marrow cells were mixed with syngenic WEHI-3B cells and this chimeric cell population was transduced with AdCD vectors. Infected cells were injected into myeloablated Balb-c mice, which then received a 5-fluorocytosine treatment for 4 days. All mice transplanted with unpurged bone marrow developed leukemia and died. However, 90% of recipients receiving the purging treatment were healthy up to 9 months post-transplantation and had a perfectly re-established hematopoietic system, without any signal of leukemic cell presence. In conclusion, suicide adenoviral vectors are proposed as a tool for the purging of Adenoviral susceptible myeloid leukemia cells contaminating autologous bone marrow grafts. PMID- 12883530 TI - Stable rAAV-mediated transduction of rod and cone photoreceptors in the canine retina. AB - Recombinant adeno-associated virus (rAAV) vectors are attractive candidates for the treatment of inherited and acquired retinal disease. Although rAAV vectors are well characterized in rodent models, a prerequisite to their clinical application in human patients is the thorough evaluation of their efficacy and safety in intermediate animal models. In this study, we describe rAAV-2-mediated expression of GFP reporter gene in retinal cells following local vector delivery in dogs. Subretinal delivery of rAAV.CMV.GFP was performed unilaterally in eight normal dogs from 6 weeks of age. The area of retinal transduction was maximized by the optimization of surgical techniques for subretinal vector delivery by pars plana vitrectomy and the use of fine-gauge subretinal cannulae to create multiple retinotomies. rAAV-2 vectors mediated efficient stable reporter gene expression in photoreceptors and retinal pigment epithelial cells. We found efficient transduction of cone photoreceptors in addition to rods in both the canine retina and after subretinal vector delivery in another intermediate animal model, the feline retina. GFP expression in dogs was confined to the area of the retinal bleb and was sustained in cells at this site for at least 18 months. Electroretinography demonstrated a modest reduction in global rod-mediated retinal function following subretinal delivery of rAAV.CMV.GFP. Three of the eight animals developed delayed-onset intraocular inflammation, in two cases associated with a serum antibody response to GFP protein. We conclude that rAAV-2 vectors mediate efficient sustained transgene expression in rod and cone photoreceptors following subretinal delivery in this intermediate animal model. The possibility of adverse effects including intraocular immune responses and reduced retinal function requires further investigation prior to clinical applications in patients. PMID- 12883531 TI - Gene therapy for new bone formation using adeno-associated viral bone morphogenetic protein-2 vectors. AB - Previous reports have suggested that bone morphogenetic protein (BMP) gene therapy could be applied for in vivo bone regeneration. However, these studies were conducted either using immunodeficient animals because of immunogenicity of adenovirus vectors, or using ex vivo gene transfer technique, which is much more difficult to handle. Adeno-associated virus (AAV) is a replication-defective virus without any association with immunogenicity and human disease. This study was conducted to investigate whether orthotopic new bone formation could be induced by in vivo gene therapy using AAV-based BMP2 vectors. To test the feasibility of this approach, we constructed an AAV vector carrying human BMP2 gene. Mouse myoblast cells (C2C12) transduced with this vector could produce and secrete biologically active BMP2 protein and induce osteogenic activity, which was confirmed by ELISA and alkaline phosphatase activity assay. For in vivo study, AAV-BMP2 vectors were directly injected into the hindlimb muscle of immunocompetent Sprague-Dawley rats. Significant new bone under X-ray films could be detected as early as 3 weeks postinjection. The ossification tissue was further examined by histological and immunohistochemical analysis. This study is, to our knowledge, the first to establish the feasibility of AAV-based BMP2 gene therapy for endochondral ossification in immunocompetent animals. PMID- 12883533 TI - Synthesis of adenoviral targeting molecules by intein-mediated protein ligation. AB - Adenoviral vectors infect cells through the binding of capsid proteins to cell surface receptors. The ubiquitous expression of adenoviral receptors in human tissues represents an obstacle toward the development of systemically deliverable vectors for cancer therapy, since effective therapy may require delivery to specific sites. For these reasons, major efforts are directed toward the elimination of the native tropism combined with identification of ligands that bind to tumor-specific cell-surface proteins. Highthroughput technologies have identified potential targeting ligands, which need to be evaluated for their ability to retarget adenovirus to alternative receptors. Here, we present a strategy that permits the routine analysis of adenoviral targeting ligands. We use intein-mediated protein ligation as a means to produce functional biological molecules, that is, adenoviral targeting molecules that function as adapters between cellular receptors and the adenovirus fiber protein. We demonstrate the versatility of the present system by conjugating targeting ligands that differ in size and nature including an apolipoprotein E synthetic peptide, the basic fibroblast growth factor and folic acid. The resulting adenoviral targeting molecules mediate adenoviral gene delivery in cells that express the corresponding receptor. PMID- 12883532 TI - Effects of two Gbetagamma-binding proteins--N-terminally truncated phosducin and beta-adrenergic receptor kinase C terminus (betaARKct)--in heart failure. AB - Myocardial overexpression of the C-terminus of beta-adrenergic receptor kinase (betaARKct) has been shown to result in a positive inotropic effect or an improvement of survival in heart failure. However, it is not clear whether this beneficial effect is mainly because of dominant-negative inhibition of betaARK1, and a consecutive resensitization of beta-adrenergic receptors (betaAR), or rather due to inhibition of other Gbetagamma-mediated effects. In this study, we tested whether overexpression of N-terminally truncated phosducin (nt-del phosducin), another Gbetagamma-binding protein that does not resensitize betaARs owing to simultaneous inhibition of GDP release from Galpha subunits, shows the same effects as betaARKct. Adenoviral gene transfer was used to express nt-del phosducin and betaARKct in isolated ventricular cardiomyocytes and in myocardium of rabbits, which suffered from heart failure because of rapid ventricular pacing. BetaAR-stimulated cAMP formation was increased by betaARKct, but not by nt-del-phosducin, whereas both proteins inhibited Gbetagamma-mediated effects. Both transgenes also increased contractility of normal and failing isolated cardiomyocytes and improved contractility in rabbits with heart failure after gene transfer in vivo. In conclusion, overexpression of nt-del-phosducin enhances the contractility of cardiomyocytes to the same extent as betaARKct, suggesting that the effects of betaARKct might be owing to inhibition of Gbetagamma rather than to betaAR resensitization. PMID- 12883534 TI - Directed evolution of retroviruses activatable by tumour-associated matrix metalloproteases. AB - Protease-activatable retroviral vectors offer the possibility of targeted gene transfer into cancer cells expressing a unique set of proteases as, for example, the matrix metalloproteases (MMPs). However, it is difficult to predict which substrate sequence will be optimally cleaved by a given tumour cell type. Therefore, we developed a novel approach that allows the selection of MMP activatable retroviruses from libraries of viruses displaying combinatorially diversified protease substrates. Starting from a virus harbouring a standard MMP 2 substrate motif, after only two consecutive cycles of diversification and in vivo selection, MMP-activatable viruses were recovered. Biochemical characterization of the selected viruses revealed that their linker peptides showed a considerably increased sensitivity for MMP-2 cleavage, and interestingly also improved the particle incorporation rate of the Env protein. Owing to the optimized linker peptide, the selected viruses exhibited a greatly enhanced spreading efficiency through human fibrosarcoma cells, while having retained the dependency on MMP activation. Moreover, cell entry efficiency and virus titres were considerably improved as compared to the parental virus displaying the standard MMP-2 substrate. The results presented imply that retroviral protease substrate libraries allow the definition of MMP substrate specificities under in vivo conditions as well as the generation of optimally adapted tumour-specific viruses. PMID- 12883535 TI - Recombinant Sendai virus vectors for activated T lymphocytes. AB - T-lymphocyte-directed gene therapy has potential as a treatment of subjects with immunological disorders. One current limitation of this therapeutic strategy is low gene transfer efficiency, even when complex procedures are used. We report herein that a recombinant Sendai virus vector (SeV) was able to overcome this issue. Using jellyfish enhanced green fluorescent protein gene (EGFP), we found that SeV was able to transduce and express a foreign gene specifically and efficiently in activated murine and human T cells, but not in naive T cells, without centrifugation or reagents including polybrene and protamine sulfate; the present findings were in clear contrast to those demonstrated with the use of retroviruses. The transduction was selective in antigen-activated T cells, while antigen-irrelevant T cells were not transduced, even under bystander activation from specific T-cell responses by antigens ex vivo. Receptor saturation studies suggested a possible mechanism of activated T-cell-specific gene transfer, ie, SeV might attach to naive T cells but might be unable to enter their cytoplasm. We therefore propose that the SeV vector system may prove to be a potentially important alternative in the area of T-cell-directed gene therapy used in the clinical setting. PMID- 12883536 TI - Adenovirus-mediated interleukin-2 gene therapy of nociception. AB - The effect of adenovirus-mediated interleukin-2 (IL-2) gene on rat basal nociceptive response and chronic neuropathic pain was explored. The paw withdrawal latency induced by radiant heat was used to evaluate the antinociceptive effect of adenovirus type 5 (Ad5) and Ad5-IL-2. The results showed that intrathecal delivery of Ad5-IL-2 exhibited obvious antinociceptive effects on basal nociceptive response and chronic neuropathic pain, which were maintained for 3 and 4 weeks, respectively. This suggested that the antinociceptive effect of Ad5-IL-2 on chronic neuropathic pain was greater than its effect on basal nociceptive response. Human IL-2 mRNA was detected by in situ hybridization in the spinal pia mater and parenchyma of the lumbar, sacral, thoracic and cervical regions, and gray matter had higher level of IL-2 expression than white matter. These data demonstrated that the IL-2 gene was transfected into spinal cord regions relevant to pain modulation. The expressed IL-2 protein profile in spinal cord detected by enzyme-linked immunosorbent assay coincided almost exactly with its antinociceptive effect. This supported the hypothesis that the therapeutic effect of IL-2 gene was related to IL-2 protein expression. The study indicates that intrathecal delivery of adenovirus-mediated IL-2 gene has a relatively long antinociceptive effect. PMID- 12883537 TI - Enhanced antitumor effect of combined replicative adenovirus and nonreplicative adenovirus expressing interleukin-12 in an immunocompetent mouse model. AB - For cancer gene therapy, replicative adenovirus is a promising vector to overcome low infectivity and poor gene delivery of nonreplicative adenovirus in vivo, but its therapeutic efficacy is still unsatisfactory because of the limited spread of replicative virus in a solid tumor. Therefore, the combined therapy with other antitumor agents may be necessary. Nonreplicative adenovirus expressing a therapeutic gene may be a promising candidate because E1 proteins expressed by replicative adenovirus would render nonreplicative adenovirus replicative, augmenting a transgene expression. In this study, we first found that mouse hepatoma Hepa 1-6 cells were permissive for the replication and cytopathic effect of human adenovirus, which enabled us to examine the potential of combined replicative adenovirus and nonreplicative adenovirus expressing an immunostimulator in an immunocompetent mouse-syngeneic Hepa 1-6 tumor model. Nonreplicative adenovirus expressing interleukin-12 (AdIL-12) was used as a model. In vitro coinfection of two adenoviruses produced higher concentrations of IL-12 than infection of AdIL-12 alone in this cell line. In vivo experiments with Hepa 1-6 tumors in syngeneic immunocompetent C57BL/6 mice showed higher concentrations of serum IL-12 and greater therapeutic efficacy in the combination therapy than infection of either adenovirus. These data indicate that the combination of replicative adenovirus and nonreplicative adenovirus expressing an immunostimulator appears to be very efficacious for cancer gene therapy. PMID- 12883538 TI - Towards therapy for spinal cord injuries. PMID- 12883539 TI - Prevention of spinal epidural fibrosis by recombinant tissue plasminogen activator in rats. AB - STUDY DESIGN: A randomized-controlled experimental study was conducted to investigate the effectiveness of recombinant tissue plasminogen activator (rt-PA) on postlaminectomy epidural fibrosis in rats. SETTING: Diyarbakir, Turkey. METHODS: In a rat model, allowing objective histopathological quantification, the postoperative epidural scar formation was investigated and the potential of local rt-PA in preventing the production of epidural fibrosis was assessed. Rats were randomly allocated to local rt-PA (12 rats) or control (12 rats). RESULTS: Findings suggest a beneficial effect of rt-PA in decreasing the epidural fibrosis following laminectomy when compared with control groups for all investigated parameters such as intermuscular scar (P=0.04), middle scar (P=0.001), deep scar (P=0.001) and dural adhesion (P=0.01) except new bone formation. The presence of arachnoiditis was less in treatment group (P=0.01). CONCLUSION: In rats, the topical thrombolysis with rt-PA is safe and efficacious in preventing postlaminectomy epidural fibrosis. This Thrombolytic therapy with rt-PA after spine surgery may come to play an important role in the prevention of epidural fibrosis and arachnoiditis. PMID- 12883540 TI - Detection of early squamous metaplasia in bladder biopsies of spinal cord injury patients by immunostaining for cytokeratin 14. AB - STUDY DESIGN: A prospective, immunohistochemical study of bladder biopsies taken from spinal cord injury (SCI) patients. OBJECTIVES: To investigate whether cytokeratin 14 immunostaining may be useful to detect early squamous metaplasia in bladder biopsies from patients with SCI. SETTING: Southport, United Kingdom. METHODS: Biopsy of bladder mucosa was taken from adults with SCI, while they underwent an elective therapeutic procedure in the urinary tract. A total of, 54 biopsies, which showed transitional epithelium only with no evidence of squamous metaplasia on routine H&E staining, formed the study group. In all, 22 biopsies, which showed squamous metaplasia on routine H&E staining, acted as controls. All biopsies were benign with no evidence of dysplasia or malignancy. Immunohistochemical staining for cytokeratin 14 was performed on all biopsies in a single batch, using a standard avidin-biotin complex method. RESULTS: All control biopsies showed positive immunostaining for cytokeratin 14 in basal and parabasal cells in areas of squamous metaplasia. Of the 54 biopsies, which showed only transitional epithelium on H&E staining, immunohistochemistry for cytokeratin 14 showed no staining in 47 biopsies. The remaining seven biopsies showed positive immunostaining for cytokeratin 14 in the epithelium, in individual cells or clusters of basal cells, revealing unexpected early squamous metaplasia in these biopsies. CONCLUSION: Immunostaining for cytokeratin 14 identifies an early phenotypic switch from transitional to squamous epithelium in bladder mucosa. Cytokeratin 14 staining is sufficiently sensitive to identify early squamous metaplasia, which is not yet evident on examination of routine H&E stained sections. This early identification may be of use in alerting physicians to change bladder management regimens to prevent predisposition to recurrent urinary infection and progression of squamous metaplasia. A cost/benefit analysis should be performed to assess the feasibility of routine cytokeratin 14 immunostaining of bladder biopsies from SCI patients. PMID- 12883541 TI - Improved kinematics of unrestrained arm raising in C5-C6 tetraplegic subjects after deltoid-to-triceps transfer. AB - STUDY DESIGN: Prospective comparative study. OBJECTIVE: To determine whether control of upper limb joint synergy during unrestrained arm raising involving shoulder and elbow flexion is modified by deltoid-to-triceps transfer. SETTING: Rehabilitation unit for spinal cord injury patients, France. METHODS: Five C6 subjects with C5-C6 tetraplegia sustained posterior deltoid-to-triceps transfer and were compared to a control group of 11 subjects. Kinematics of shoulder and elbow joints before, 6 month and 1 year after surgery, during straight-arm raising (SAR) in the plane of the scapula and hand-to-nape-of-neck movements (HNNMs) were explored. Motion was recorded with a six-camera Vicon motion analysis system and the data used to assess the coupling of elbow and shoulder flexion velocities (EFVs and SFVs) and extension velocities. RESULTS: All subjects were initially assessed at 6.9 months (mean) postsurgery. Three of the upper limbs were assessed a second time (mean 17.9 months). The first assessment showed an increase in shoulder flexion amplitudes in tetraplegic subjects with presurgery shoulder flexion deficits. Peak SFVs and EFVs and extension velocities were slightly modified at the first postsurgery assessment and dramatically improved at the second assessment during both SAR and HNNM. Despite these increased velocities, joint coordination was only partially restored. CONCLUSION: A restored elbow active extension improves the speed of elbow flexion. The additional improvement of shoulder motion emphasises the relation between joints in the control of arm movement. PMID- 12883542 TI - Maintenance of exercise participation in individuals with spinal cord injury: effects on quality of life, stress and pain. AB - STUDY DESIGN: Follow-up study of seven individuals with spinal cord injury (SCI) who completed a 9-month randomized control trial (RCT) of exercise training. OBJECTIVE: In a 9-month RCT conducted in our lab, individuals with SCI who participated in a twice-weekly supervised exercise training reported greater perceived quality of life (PQOL), and less stress and pain than a nonexercising control cohort. The present follow-up study examined the voluntary continuation of exercise training after the study ended and the persistence of the accrued psychosocial benefits. SETTING: Centre for Health Promotion and Rehabilitation, McMaster University, Hamilton, Ontario, Canada. METHODS: Five men and two women (age 42.3+/-3.6 years) with SCI (C5-T12; ASIA A-D 12.7+/-8.2 years postinjury) were invited to continue supervised exercise training twice weekly at the completion of the 9-month RCT. Exercise adherence, PQOL, stress and bodily pain were measured at a 3-month follow-up and were compared to values obtained at baseline, and at 3, 6 and 9 months during the intervention. RESULTS: There was a significant decrease in adherence at the 3-month follow-up compared to the overall 9-month adherence rate (42.7 versus 80.6%, respectively; P<0.01). There was also a significant decrease in PQOL (P<0.05) and a trend for increased pain (P=0.07) and stress (P=0.12), at follow-up compared to the end of the 9-month trial. Finally, there was a significant negative correlation between pain at the conclusion of the RCT and exercise adherence over the 3-month follow-up period (r=-0.91; P<0.01). CONCLUSIONS: These findings emphasize the importance of continued exercise adherence to the maintenance of exercise-related increases in psychological well-being among individuals with SCI. PMID- 12883543 TI - Effects of a wheelchair ergometer training programme on spinal cord-injured persons. AB - STUDY DESIGN: Before and after investigation of the effects of a wheelchair ergometer Training programme. OBJECTIVE: To investigate the effects of an original interval-training programme on work capacity and cardiorespiratory variables with spinal cord-injured persons (SCIP's) on a wheelchair-specific ergometer. SETTING: BESANCON, FRANCE. METHODS: Seven SCIP's (male) performed 45 min of wheelchair ergometry three times per week, for 6 weeks. Training effects on maximal dynamic performance and endurance capacity were studied by comparison of performance and cardiorespiratory responses observed during both a maximal progressive test (10 W/2 min) and the same training session performed before and after training. RESULTS: Training induced significant improvements in maximal tolerated power (+19.6%), in peak oxygen consumption (VO2,+16%), and in oxygen pulse (O2p,+18.7%). At ventilatory threshold, significant improvements were also observed in power output (+63%), VO2VT(+ 34.1), ventilation VEVT(+ 37.1%), and V2pVT(+ 19.9% ). Heart rate and ventilation were significantly lower (-11 and 14.6%, respectively) after training at the same work rate, while VO2 was unchanged. Between the first and the last training session, the total physical work was improved by 24.7%, whereas heart rate was unchanged. CONCLUSION: An interval-training programme individualised to each paraplegic subject using a wheelchair ergometer can significantly improve the fitness level and endurance capacity. PMID- 12883544 TI - Effects on age on spinal cord lesion patients' rehabilitation. AB - STUDY DESIGN: The present study was undertaken to focus the age-related characteristics of a population of traumatic and nontraumatic spinal cord patients. OBJECTIVES: to examine demographic, injury and outcome characteristics of older adults with spinal cord lesions as a result of trauma and nontrauma, and to compare these characteristics with those of younger patients in matched cohorts. SETTING: Spinal Cord Unit, Fondazione Santa Lucia IRCCS, a large rehabilitation hospital of the centre-south of Italy. METHODS: In total, 284 consecutive newly injured patients with traumatic and nontraumatic spinal cord lesions were retrospectively reviewed and divided according to age into two groups: under 50 years (group 1) and over 50 years (group 2). The following information was collected: onset of lesion to admission; injury variables: aetiology, level, associated injuries, medical complications and surgical intervention; length of stay; American Spinal Injury Association (ASIA) impairment and motor scores; Barthel Index (BI) and Rivermead Mobility Index (RMI) to assess independence in daily living; Walking Index for Spinal Cord Injury to assess ambulation; patients destination at discharge. In a subset of 130 subjects, a block design, matching procedure was used to control for the covariant effects of injury characteristics, time from lesion and aetiology on age effects. RESULTS: In the entire group of 284 patients, older subjects had a higher probability of having incomplete tetraplegia of nontraumatic origin; they also showed a shorter length of stay and a higher rate of complications. In the matched cohorts, younger patients showed better neurologic recovery (intended as ASIA impairment grade improvement and motor scores increase), significantly higher Barthel Index and RMI at discharge, a higher level of independence in spontaneous bladder and bowel management and a higher frequency of independent walking. CONCLUSION: Older individuals with spinal cord injury and disease do well, but have a less favourable outcome in regard to walking, bladder and bowel independence than younger subjects and have more associated medical problems. Different rehabilitative strategies, therefore, are required for older subjects, which maximises the shorter length of stay and provides the necessary medical care and increased physical assistant resources following discharge. PMID- 12883545 TI - Use of a latissimus dorsi flap for treatment of infection in a neuropathic shoulder joint. AB - STUDY DESIGN: Case report. OBJECTIVES: To demonstrate the difficulties that can be encountered in diagnosis and treatment of an infected neuropathic shoulder in a paraplegic man. SETTING: Spinal cord injury center in Germany. METHOD: Ultimately, radical debridement and transplantation of a latissimus dorsi muscle flap. RESULTS: Successful treatment of the infection, partial weight bearing, and limited restriction of range of motion in the affected shoulder. CONCLUSION: The latissimus dorsi muscle flap can resolve the infection of a neuropathic shoulder. PMID- 12883546 TI - Spontaneous resolution of lumbar pseudomeningocoele. AB - STUDY DESIGN: A case report of spontaneous resolution of a lumbar postdiscectomy pseudomeningocoele. OBJECTIVES: To suggest the role of nonoperative treatment even in symptomatic pseudomeningocoeles. SETTING: Withybush General Hospital, Haverfordwest, Pembrokeshire, South Wales, UK. CASE REPORT: A 65-year-old lady underwent L4/L5 discectomy for lumbar disc prolapse in 1998. As the patient did not have relief of symptoms, an MRI was taken at 1 month following the operation, which showed a residual disc at L4/L5 and a pseudomeningocoele communicating with the subarachnoid space. The patient could not undergo further treatment because of the untimely demise of the surgeon. Over the next 3 months, the symptoms began to improve and the patient was totally asymptomatic and remained so for 3 years. In 2001, she was seen for a recurring leg pain and back pain and an MRI was done, which showed complete disappearance of the pseudomeningocoele but with recurrent disc lesion. CONCLUSION: Although the current medical literature favours re exploration and repair of the dural defect in symptomatic pseudomeningocoele, the authors are of the opinion that conservative treatment may have a role in the treatment of the above condition as illustrated by the above example. PMID- 12883547 TI - Anterior spinal artery syndrome after peripheral angiography of the lower limbs. AB - STUDY DESIGN: A case report of temporary anterior spinal artery syndrome secondary to peripheral angiography of the lower limbs. OBJECTIVES: To report a rare case of spinal cord ischemia following a straightforward peripheral angiography with femoral access. SETTING: Istanbul, Turkey. CASE REPORT: A 70 year-old patient with peripheral vascular disease developed temporary paraplegia after angiography caused by anterior spinal ischemia. His motor weakness improved gradually over a 2 month period. CONCLUSION: Spinal complications are extremely rare after peripheral angiography. The possible mechanisms of spinal cord damage are discussed, and the relevant data regarding the etiology are reviewed. PMID- 12883548 TI - Should spinal cord clinicians be proactive in preventing spinal cord injuries and decreasing secondary complications caused by spinal cord injury? PMID- 12883549 TI - The remarkable difference between surface and step atoms in the magnetic anisotropy of two-dimensional nanostructures. AB - The original magnetic properties of nanometre-sized particles are due to the distinct contributions of volume, surface and step atoms. To disentangle these contributions is an ongoing challenge of materials science. Here we introduce a method enabling the identification of the remarkably different contributions of surface and perimeter atoms to the magnetic anisotropy energy of two-dimensional nanostructures. Our method uses the generally nonlinear relationship between perimeter length and surface area. Atomic-scale characterization of the morphology of ensembles of polydisperse nanostructures, combined with in situ measurements of their temperature-dependent magnetic susceptibility, gives access to the role played by the differently coordinated atoms. We show for Co nanostructures on a Pt(111) surface that their uniaxial out-of-plane magnetization is entirely caused by edge atoms having 20 times more anisotropy energy than their bulk and surface counterparts. Identification of the role of perimeter and surface atoms opens up unprecedented opportunities for materials engineering. As an example, we separately tune magnetic hardness and moment in bimetallic core-shell nanostructures. PMID- 12883550 TI - Thermoelectric microdevice fabricated by a MEMS-like electrochemical process. AB - Microelectromechanical systems (MEMS) are the basis of many rapidly growing technologies, because they combine miniature sensors and actuators with communications and electronics at low cost. Commercial MEMS fabrication processes are limited to silicon-based materials or two-dimensional structures. Here we show an inexpensive, electrochemical technique to build MEMS-like structures that contain several different metals and semiconductors with three-dimensional bridging structures. We demonstrate this technique by building a working microthermoelectric device. Using repeated exposure and development of multiple photoresist layers, several different metals and thermoelectric materials are fabricated in a three-dimensional structure. A device containing 126 n-type and p type (Bi, Sb)2Te3 thermoelectric elements, 20 microm tall and 60 microm in diameter with bridging metal interconnects, was fabricated and cooling demonstrated. Such a device should be of technological importance for precise thermal control when operating as a cooler, and for portable power when operating as a micro power generator. PMID- 12883551 TI - Switching of magnetization by nonlinear resonance studied in single nanoparticles. AB - Magnetization reversal in magnetic particles is one of the fundamental issues in magnetic data storage. Technological improvements require the understanding of dynamical magnetization reversal processes at nanosecond time scales. New strategies are needed to overcome current limitations. For example, the problem of thermal stability of the magnetization state (superparamagnetic limit) can be pushed down to smaller particle sizes by increasing the magnetic anisotropy. High fields are then needed to reverse the magnetization, which are difficult to achieve in current devices. Here we propose a new method to overcome this limitation. A constant applied field, well below the switching field, combined with a radio-frequency (RF) field pulse can reverse the magnetization of a nanoparticle. The efficiency of this method is demonstrated on a 20-nm-diameter cobalt particle by using the microSQUID (superconducting quantum interference device) technique. Other applications of this method might be nucleation or depinning of domain walls. PMID- 12883552 TI - Enhanced T cell responses due to diacylglycerol kinase zeta deficiency. AB - Much is known about how T cell receptor (TCR) engagement leads to T cell activation; however, the mechanisms terminating TCR signaling remain less clear. Diacylglycerol, generated after TCR ligation, is essential in T cells. Its function must be controlled tightly to maintain normal T cell homeostasis. Previous studies have shown that diacylglycerol kinase zeta (DGKzeta), which converts diacylglycerol to phosphatidic acid, can inhibit TCR signaling. Here we show that DGKzeta-deficient T cells are hyperresponsive to TCR stimulation both ex vivo and in vivo. Furthermore, DGKzeta-deficient mice mounted a more robust immune response to lymphocytic choriomeningitis virus infection than did wild type mice. These results demonstrate the importance of DGKzeta as a physiological negative regulator of TCR signaling and T cell activation. PMID- 12883553 TI - Small-conductance, Ca2+-activated K+ channel SK3 generates age-related memory and LTP deficits. AB - Cognitive deficits are among the most devastating changes associated with the aging process. Age-related decrement in performance on learning tasks is correlated with substantial changes in neuronal signal processing in the hippocampus. Here we show that elevated expression of small-conductance Ca2+ activated K+ channels (SK channels) of the SK3 type in hippocampi of aged mice contributes to reduced long-term potentiation (LTP) and impaired trace fear conditioning, a hippocampus-dependent learning task. PMID- 12883554 TI - A non-proteolytic role for ubiquitin in Tat-mediated transactivation of the HIV-1 promoter. AB - The human immunodeficiency virus type 1 (HIV-1) encodes a potent transactivator, Tat, which functions through binding to a short leader RNA, called transactivation responsive element (TAR). Recent studies suggest that Tat activates the HIV-1 long terminal repeat (LTR), mainly by adapting co-activator complexes, such as p300, PCAF and the positive transcription elongation factor P TEFb, to the promoter. Here, we show that the proto-oncoprotein Hdm2 interacts with Tat and mediates its ubiquitination in vitro and in vivo. In addition, Hdm2 is a positive regulator of Tat-mediated transactivation, indicating that the transcriptional properties of Tat are stimulated by ubiquitination. Fusion of ubiquitin to Tat bypasses the requirement of Hdm2 for efficient transactivation, supporting the notion that ubiquitin has a non-proteolytic function in Tat mediated transactivation. PMID- 12883555 TI - Utilization of the BackMed email discussion list in a specialized health sciences learning center: a cost-benefit analysis. PMID- 12883556 TI - What is the best way to gather clinical questions from physicians? PMID- 12883557 TI - Sharing the burden: a model for consortium purchasing for health libraries. PMID- 12883558 TI - The role of the academic medical center library in training public librarians. AB - PURPOSE: This project enhanced access to and awareness of health information resources on the part of public libraries in western Pennsylvania. SETTING/PARTICIPANTS/RESOURCES: The Health Sciences Library System (HSLS), University of Pittsburgh, conducted a needs assessment and offered a series of workshops to 298 public librarians. BRIEF DESCRIPTION: The National Library of Medicine-funded project "Access to Electronic Health Information" at the HSLS, University of Pittsburgh, provided Internet health information training to public libraries and librarians in sixteen counties in western Pennsylvania. Through this project, this academic medical center library identified the challenges for public librarians in providing health-related reference service, developed a training program to address those challenges, and evaluated the impact of this training on public librarians' ability to provide health information. RESULTS/OUTCOME: The HSLS experience indicates academic medical center libraries can have a positive impact on their communities by providing health information instruction to public librarians. The success of this project--demonstrated by the number of participants, positive course evaluations, increased comfort level with health-related reference questions, and increased use of MEDLINEplus and other quality information resources--has been a catalyst for continuation of this programming, not only for public librarians but also for the public in general. EVALUATION METHOD: A training needs assessment, course evaluation, and impact training survey were used in developing the curriculum and evaluating the impact of this training on public librarians' professional activities. PMID- 12883559 TI - Current issues in the design of academic health sciences libraries: findings from three recent facility projects. AB - Planning a new health sciences library at the beginning of the twenty-first century is a tremendous challenge. Technology has radically changed the way libraries function in an academic environment and the services they provide. Some individuals question whether the library as place will continue to exist as information becomes increasingly available electronically. To understand how libraries resolve programming and building design issues, visits were made to three academic health sciences libraries that have had significant renovation or completed new construction. The information gathered will be valuable for planning a new library for the University of Colorado Health Sciences Center and may assist other health sciences librarians as they plan future library buildings. PMID- 12883560 TI - Identifying diagnostic accuracy studies in EMBASE. AB - OBJECTIVE: The objective was to develop and test search strategies to identify diagnostic articles recorded on EMBASE. METHODS: Four general medical journals were hand searched for diagnostic accuracy studies published in 1999. Identified studies served as a gold standard. Candidate terms for search strategies were identified using a word-frequency analysis of their abstracts. According to the frequency of identified terms, searches were run for each term independently. Sensitivity, precision, and number needed to read (NNR) (1/precision) of every candidate term were calculated. Terms with the highest "sensitivity*precision" product were used as free-text terms and combined into a final strategy using the Boolean operator "OR." RESULTS: The most frequently occurring eight terms (sensitiv* or detect* or accura* or specific* or reliab* or positive or negative or diagnos*) produced a sensitivity of 100% (95% confidence interval [CI] 94.1 to 100%) and an NNR of 27 (95% CI 21.0 to 34.8). The combination of the two truncated terms sensitiv* or detect* gave a sensitivity of 73.8% (95% CI 60.9 to 84.2%) and an NNR of 5.7 (95% CI 4.4 to 7.6). CONCLUSIONS: The identified search terms offer the choice of either reasonably sensitive or precise search strategies for the detection of diagnostic accuracy studies in EMBASE. The terms are useful both for busy health care professionals who value precision and for reviewers who value sensitivity. PMID- 12883561 TI - Family physicians' interests in special features of electronic publication. AB - OBJECTIVE: Because many of the medical journals read by family physicians now have an electronic version, the authors conducted a survey to determine the interest of family physicians in specific features of electronic journal publications. SETTING AND PARTICIPANTS: We surveyed 175 family physicians randomly selected from the American Academy of Family Physicians. RESULTS: The response rate was 63%. About half of family physicians reported good to excellent computer proficiency, and about one quarter used online journals sometimes or often. Many respondents reported high interest in having links to: an electronic medical text (48% for original articles, 56% for review articles), articles' list of references (52% for original articles, 56% for review articles), and health related Websites (48% for original and review articles). CONCLUSION: Primary care oriented journals should consider the interests of family physicians when developing and offering electronic features for their readers. PMID- 12883562 TI - Benchmarking information needs and use in the Tennessee public health community. AB - OBJECTIVE: The objective is to provide insight to understanding public health officials' needs and promote access to data repositories and communication tools. METHODS: Survey questions were identified by a focus group with members drawn from the fields of librarianship, public health, and informatics. The resulting comprehensive information needs survey, organized in five distinct broad categories, was distributed to 775 Tennessee public health workers from ninety five counties in 1999 as part of the National Library of Medicine-funded Partners in Information Access contract. RESULTS: The assessment pooled responses from 571 public health workers (73% return rate) representing seventy-two of ninety-five counties (53.4% urban and 46.6% rural) about their information-seeking behaviors, frequency of resources used, computer skills, and level of Internet access. Sixty four percent of urban and 43% of rural respondents had email access at work and more than 50% of both urban and rural respondents had email at home (N = 289). Approximately 70% of urban and 78% of rural public health officials never or seldom used or needed the Centers for Disease Control (CDC) Website. Frequency data pooled from eleven job categories representing a subgroup of 232 health care professionals showed 72% never or seldom used or needed MEDLINE. Electronic resources used daily or weekly were email, Internet search engines, internal databases and mailing lists, and the Tennessee Department of Health Website. CONCLUSIONS: While, due to the small sample size, data cannot be generalized to the larger population, a clear trend of significant barriers to computer and Internet access can be identified across the public health community. This contributes to an overall limited use of existing electronic resources that inhibits evidence-based practice. PMID- 12883563 TI - Information-seeking behavior in complementary and alternative medicine (CAM): an online survey of faculty at a health sciences campus. AB - BACKGROUND: The amount of reliable information available for complementary and alternative medicine (CAM) is limited, and few authoritative resources are available. OBJECTIVE: The objective is to investigate the information-seeking behavior of health professionals seeking CAM information. METHODS: Data were gathered using a Web-based questionnaire made available to health sciences faculty affiliated with the University of California, San Francisco. RESULTS: The areas of greatest interest were herbal medicine (67%), relaxation exercises (53%), and acupuncture (52%). About half the respondents perceived their CAM searches as being only partially successful. Eighty-two percent rated MEDLINE as a useful resource, 46% personal contacts with colleagues, 46% the Web, 40% journals, and 20% textbooks. Books and databases most frequently cited as useful had information about herbs. The largest group of respondents was in internal medicine (26%), though 15% identified their specialties as psychiatry, psychology, behavioral medicine, or addiction medicine. There was no correlation between specialty and patterns of information-seeking behavior. Sixty-six percent expressed an interest in learning more about CAM resources. CONCLUSIONS: Health professionals are frequently unable to locate the CAM information they need, and the majority have little knowledge of existing CAM resources, relying instead on MEDLINE. Medical librarians need to educate health professionals in the identification and use of authoritative CAM resources. PMID- 12883564 TI - Mapping the literature of clinical laboratory science. AB - This paper describes a citation analysis of the literature of clinical laboratory science (medical technology), conducted as part of a project of the Nursing and Allied Health Resources Section of the Medical Library Association. Three source journals widely read by those in the field were identified, from which cited references were collected for a three-year period. Analysis of the references showed that journals were the predominant format of literature cited and the majority of the references were from the last eleven years. Applying Bradford's Law of Scattering to the list of journals cited, three zones were created, each producing approximately one third of the cited references. Thirteen journals were in the first zone, eighty-one in the second, and 849 in the third. A similar list of journals cited was created for four specialty areas in the field: chemistry, hematology, immunohematology, and microbiology. In comparing the indexing coverage of the Zone 1 and 2 journals by four major databases, MEDLINE provided the most comprehensive coverage, while the Cumulative Index to Nursing and Allied Health Literature was the only database that provided complete coverage of the three source journals. However, to obtain complete coverage of the field, it is essential to search multiple databases. PMID- 12883565 TI - Developing an academic medical library core journal collection in the (almost) post-print era: the Florida State University College of Medicine Medical Library experience. AB - The Florida State University (FSU) College of Medicine Medical Library is the first academic medical library to be established since the Web's dramatic appearance during the 1990s. A large customer base for electronic medical information resources is both comfortable with and eager to migrate to the electronic format completely, and vendors are designing radical pricing models that make print journal cancellations economically advantageous. In this (almost) post-print environment, the new FSU Medical Library is being created and will continue to evolve. By analyzing print journal subscription lists of eighteen academic medical libraries with similar missions to the community-based FSU College of Medicine and by entering these and selected quality indicators into a Microsoft Access database, a core list was created. This list serves as a selection guide, as a point for discussion with faculty and curriculum leaders when creating budgets, and for financial negotiations in a broader university environment. After journal titles specific to allied health sciences, veterinary medicine, dentistry, pharmacy, library science, and nursing were eliminated from the list, 4,225 unique journal titles emerged. Based on a ten-point scale including SERHOLD holdings and DOCLINE borrowing activity, a list of 449 core titles is identified. The core list has been saved in spreadsheet format for easy sorting by a number of parameters. PMID- 12883566 TI - Patricia L. Thibodeau Medical Library Association President 2003-2004. PMID- 12883567 TI - More concerns about impact factors. PMID- 12883568 TI - Challenges for libraries creating one world: information ethics and policy issues for medical librarians. PMID- 12883569 TI - Whose business is it? PMID- 12883570 TI - The Leadership Reconsidered symposium: report. PMID- 12883571 TI - Librarians as contributing authors to Patient Education and Preventive Medicine: a collaborative project of the Consumer Health Committee of the Georgia Health Sciences Library Association. PMID- 12883572 TI - Exploring new ways of publishing: a library-faculty partnership. PMID- 12883573 TI - Controlling hospital library theft. AB - At Capital Health System/Fuld Campus (formerly Helene Fuld Medical Center), the Health Sciences Library lost many books and videocassettes. These materials were listed in the catalog but were missing when staff went to the shelves. The hospital had experienced a downsizing of staff, a reorganization, and a merger. When the library staff did an inventory, $10,000 worth of materials were found to be missing. We corrected the situation through a series of steps that we believe will help other libraries control their theft. Through regularly scheduling inventories, monitoring items, advertising, and using specific security measures, we have successfully controlled the library theft. The January 2002 inventory resulted in meeting our goal of zero missing books and videocassettes. We work to maintain that goal. PMID- 12883574 TI - Measuring use patterns of online journals and databases. AB - PURPOSE: This research sought to determine use of online biomedical journals and databases and to assess current user characteristics associated with the use of online resources in an academic health sciences center. SETTING: The Library of the Health Sciences-Peoria is a regional site of the University of Illinois at Chicago (UIC) Library with 350 print journals, more than 4,000 online journals, and multiple online databases. METHODOLOGY: A survey was designed to assess online journal use, print journal use, database use, computer literacy levels, and other library user characteristics. A survey was sent through campus mail to all (471) UIC Peoria faculty, residents, and students. RESULTS: Forty-one percent (188) of the surveys were returned. Ninety-eight percent of the students, faculty, and residents reported having convenient access to a computer connected to the Internet. While 53% of the users indicated they searched MEDLINE at least once a week, other databases showed much lower usage. Overall, 71% of respondents indicated a preference for online over print journals when possible. CONCLUSIONS: Users prefer online resources to print, and many choose to access these online resources remotely. Convenience and full-text availability appear to play roles in selecting online resources. The findings of this study suggest that databases without links to full text and online journal collections without links from bibliographic databases will have lower use. These findings have implications for collection development, promotion of library resources, and end-user training. PMID- 12883575 TI - 1927 reference in the new millennium: where is the Automat? AB - James Ballard, director at the Boston Medical Library, tracked questions he received at the reference desk in 1927 to recognize the trend of queries and to record the information for future use. He presented a paper on reference services that listed sixty of his reference questions at the Thirtieth Annual Meeting of the Medical Library Association (MLA) in 1927. During a two-month period in 2001, the authors examined Ballard's questions by attempting to answer them with print sources from the 1920s and with the Internet. The searchers answered 85% of the questions with the Internet and 80% with 1920s reference sources. The authors compared Internet and 1920s print resources for practical use. When answering the questions with 1920s resources, the searchers rediscovered a time in health sciences libraries when there was no Ulrich's Periodicals Directory, no standardized subject headings, and no comprehensive listings of available books. Yet, the authors found many of the 1920s reference materials to be quite useful and often multifunctional. The authors recorded observations regarding the impact of automation on answering reference questions. Even though the Internet has changed the outward appearance of reference services, many things remain the same. PMID- 12883576 TI - The use of online information resources by nurses. AB - PURPOSE: Based on the results of an informal needs assessment, the Usage of Online Information Resources by Nurses Project was designed to provide clinical nurses with accurate medical information at the point of care by introducing them to existing online library resources through instructional classes. Actual usage of the resources was then monitored for a set period of time. METHODS: A two-hour hands-on class was developed for interested nurses. Participants were instructed in the content and use of several different online resources. A special Web page was designed for this project serving as an access point to the resources. Using a password system and WebTrends trade mark software, individual participant's usage of the resources was monitored for a thirty-day period following the class. At the end of the thirty days, usage results were tabulated, and participants were sent general evaluation forms. RESULTS: Eight participants accessed the project page thirty-nine times in a thirty-day period. The most accessed resource was Primary Care Online (PCO), accessed thirty-three times. PCO was followed by MD Consult (17), Ovid (8), NLM resources (5), and electronic journals (1). The individual with the highest usage accessed the project page thirteen times. CONCLUSIONS: Practicing clinical nurses will use online medical information resources if they are first introduced to them and taught how to access and use them. Health sciences librarians can play an important role in providing instruction to this often overlooked population. PMID- 12883577 TI - Information needs and information seeking in primary care: a study of nurse practitioners. AB - OBJECTIVE: The objective was to understand the information-related behavior of nurse practitioners (NPs), a population of clinicians responsible for an increasing proportion of primary care. METHODS: Two phases of data collection addressed seven research questions. The initial phase of data collection was a questionnaire sent to 300 NPs, who were asked to report their experiences of needing information as a result of patient encounters as well as their experiences of seeking information. The second phase of data collection entailed a series of interviews with twenty NPs following their encounters with patients to collect data on instances of information needs and information seeking. RESULTS: NPs most frequently needed information related to drug therapy and diagnosis. NPs with a master's degree were found to perceive information needs more frequently than their colleagues who had not received a master's degree. The information resources NPs used most frequently were consultations with colleagues, drug reference manuals, and textbooks and protocol manuals. NPs were more likely to pursue needs related to drug therapy with a print resource and needs related to diagnosis with a colleague. The generalizability of a need emerged as a negative predictor of information seeking. CONCLUSIONS: This study has addressed a number of questions about the information-related behavior of NPs in primary care practices and led to the development of a temporal model of information seeking in these settings. Results of this research underscore the importance of access to information resources in primary care practices. This study's findings also support the development of educational and outreach programs to promote evidence-based decision making among primary care clinicians. PMID- 12883578 TI - The Association of Academic Health Sciences Libraries Annual Statistics: an exploratory twenty-five-year trend analysis. AB - This paper presents an exploratory trend analysis of the statistics published over the past twenty-four editions of the Annual Statistics of Medical School Libraries in the United States and Canada. The analysis focuses on the small subset of nineteen consistently collected data variables (out of 656 variables collected during the history of the survey) to provide a general picture of the growth and changing dimensions of services and resources provided by academic health sciences libraries over those two and one-half decades. The paper also analyzes survey response patterns for U.S. and Canadian medical school libraries, as well as osteopathic medical school libraries surveyed since 1987. The trends show steady, but not dramatic, increases in annual means for total volumes collected, expenditures for staff, collections and other operating costs, personnel numbers and salaries, interlibrary lending and borrowing, reference questions, and service hours. However, when controlled for inflation, most categories of expenditure have just managed to stay level. The exceptions have been expenditures for staff development and travel and for collections, which have both outpaced inflation. The fill rate for interlibrary lending requests has remained steady at about 75%, but the mean ratio of items lent to items borrowed has decreased by nearly 50%. PMID- 12883579 TI - The Association of Academic Health Sciences Libraries Annual Statistics: a thematic history. AB - The Annual Statistics of Medical School Libraries in the United States and Canada (Annual Statistics) is the most recognizable achievement of the Association of Academic Health Sciences Libraries in its history to date. This article gives a thematic history of the Annual Statistics, emphasizing the leadership role of editors and Editorial Boards, the need for cooperation and membership support to produce comparable data useful for everyday management of academic medical center libraries and the use of technology as a tool for data gathering and publication. The Annual Statistics' origin is recalled, and survey features and content are related to the overall themes. The success of the Annual Statistics is evident in the leadership skills of the first editor, Richard Lyders, executive director of the Houston Academy of Medicine-Texas Medical Center Library. The history shows the development of a survey instrument that strives to produce reliable and valid data for a diverse group of libraries while reflecting the many complex changes in the library environment. The future of the Annual Statistics is assured by the anticipated changes facing academic health sciences libraries, namely the need to reflect the transition from a physical environment to an electronic operation. PMID- 12883580 TI - Tracing technology in the Association of Academic Health Sciences Libraries. AB - From the beginning of the association, technology and the Association of Academic Health Sciences Libraries (AAHSL) have been intertwined. Technology was the focus of one of the first committees. Innovative applications of technology have been employed in the operations of the association. Early applications of mini computers were used in preparing the Annual Statistics. The association's use of network communications was among the first in the country and later applications of the Web have enhanced association services. For its members, technology has transformed libraries. The association's support of the early development of Integrated Advanced Information Management Systems (IAIMS) and of its recent reconceptualization has contributed to the intellectual foundation for this revolution. PMID- 12883581 TI - The Association of Academic Health Sciences Libraries' legislative activities and the Joint Medical Library Association/Association of Academic Health Sciences Libraries Legislative Task Force. AB - The Association of Academic Health Sciences Libraries' (AAHSL's) involvement in national legislative activities and other advocacy initiatives has evolved and matured over the last twenty-five years. Some activities conducted by the Medical Library Association's (MLA's) Legislative Committee from 1976 to 1984 are highlighted to show the evolution of MLA's and AAHSL's interests in collaborating on national legislative issues, which resulted in an agreement to form a joint legislative task force. The history, work, challenges, and accomplishments of the Joint MLA/AAHSL Legislative Task Force, formed in 1985, are discussed. PMID- 12883582 TI - The Association of Academic Health Sciences Libraries' collaboration with the Association of American Medical Colleges, Medical Library Association, and other organizations. AB - The Association of Academic Health Sciences Libraries has made collaboration with other organizations a fundamental success strategy throughout its twenty-five year history. From the beginning its relationships with Association of American Medical Colleges and with the Medical Library Association have shaped its mission and influenced its success at promoting academic health sciences libraries' roles in their institutions. This article describes and evaluates those relationships. It also describes evolving relationships with other organizations including the National Library of Medicine and the Association of Research Libraries. PMID- 12883583 TI - A greater voice for academic health sciences libraries: the Association of Academic Health Sciences Libraries' vision. AB - The founders of the Association of Academic Health Sciences Libraries (AAHSL) envisioned the development of a professional organization that would provide a greater voice for academic health sciences libraries, facilitate cooperation and communication with the Association of American Medical Colleges, and create a forum for identifying problems and solutions that are common to academic health sciences libraries. This article focuses on the fulfillment of the "greater voice" vision by describing action and leadership by AAHSL and its members on issues that directly influenced the role of academic health sciences libraries. These include AAHSL's participation in the work that led to the publication of the landmark report, Academic Information in the Academic Health Sciences Center: Roles for the Library in Information Management; its contributions to the recommendations of the Physicians for the Twenty-first Century: The GPEP Report; and the joint publication with the Medical Library Association of Challenge to Action: Planning and Evaluation Guidelines for Academic Health Sciences Libraries. PMID- 12883584 TI - Present at the creation: the founding and formative years of the Association of Academic Health Sciences Libraries. AB - The Association of Academic Health Sciences Libraries (AAHSL) was founded in 1978 with the goal of strengthening academic health sciences libraries and increasing their participation nationally in efforts to improve medical education. A primary objective of the organization was to achieve a formal relationship with the Association of American Medical Colleges (AAMC) through membership in the Council of Academic Societies (CAS). Initial steps in establishing AAHSL are examined, including its efforts to join CAS. The author pays tribute to AAHSL's founders, in particular Gerald Oppenheimer, without whose vision and leadership AAHSL would not have been formed. PMID- 12883587 TI - A rose is a rose. PMID- 12883588 TI - Friends and colleagues. PMID- 12883590 TI - [The liaison psychiatry approach of the psychiatric crisis, urgencies and emergencies]. AB - The purpose of this paper is to differentiate crisis, emergencies and urgencies within the frame of Liaison psychiatry. It begins with the definition of each one of the terms, later the emphasis is put in the clinical characteristics of each one of these situations. These characteristics are determined by the patient and the therapeutic team. At last therapeutic guidelines are stated, which allow more precision in the intervention, in function of the direct involvement of these situations in the development and evolution of the patients. PMID- 12883589 TI - [Randomized, open label study on the use of ranitidine at different doses for the management of weight gain associated with olanzapine administration]. AB - Potential weight gain must be taken into account in the patient's comprehensive treatment approach when initiating antipsychotic treatment with Olanzapine. There are effective preventive and therapeutic pharmacological interventions to control this adverse effect. Not addressing this eventual treatment aspect of Olanzapine treatment may compromise the patient's health and his/her compliance with the pharmacological treatment indicated in serious psychiatric conditions. The decision to use a molecule must be made taking into account its efficacy, efficiency and adverse effect profile. In this study, concomitant administration of Ranitidine prevented or corrected weight gain in 59.6% of cases. Patients followed by 16 weeks had shown the following results: Olanzapine without Ranitidine, exhibited an average weight gain of 3.4 kilograms, ranging between 2.5 and +16 kg. This implies an average increase of 1.19 in BMI for this group. Patients treated additionally with Ranitidine at doses of 300 mg, a 0.9 kilogram weight gain ranging between -4 and +10.6 kg was observed, implying an average BMI change of 0.34. In patients treated with Ranitidine at doses of 600 mg, the weight gain curve trended toward normalization with a 1.6 kilogram decrease, ranging between -15 and +7 kilograms, accounting for a decrease of 0.6 points in BMI. While more extensive studies are required, our interest with this study is to demonstrate that we can count on a potentially simple and useful response for the treatment of weight gain associated with Olanzapine use. The risk/benefit ratio is a paradigm in the practice of medicine. In the case of Olanzapine, the scale clearly tips in favor of the latter, making it the drug of choice in the treatment of schizophrenia and other psychotic disorders. PMID- 12883591 TI - [Group therapy of bulimia nervosa and other related disorders]. AB - The aim of this article is to present a brief group intervention model for the initial phase of the treatment of bulimia nervosa and other related eating disorders. The model combines elements from cognitive-behavioral and psychoeducational approaches. Along the article we will describe the theoretical foundations of the model, a review of the empirical evidence that supports it, and afterwards, the group procedure developed by us in our clinical experience. Later on, the clinical results of an uncontrolled study of a sample of patients who participated in the group will be introduced. Finally, we will discuss the scope and limitations of this experience. PMID- 12883592 TI - [Suicide attempts in elderly people. Data from the Hospital Italiano de Buenos Aires]. AB - OBJECTIVE: [corrected] The aim of this paper is to analyze some characteristics of 25 patients, over 60, who attempted suicide and were hospitalized in the Department of Psychiatry of a general hospital in Buenos Aires. METHODS: This is a retrospective study using data from the medical records of patients (1999 to 2002); based on a protocol comprising an analysis of attempted suicides, demographic and clinical variables. Patients were diagnosed following the DSM-IV criteria by two trained GPs and were confirmed by MMPI and Rorschach. RESULTS: 72% were women. The average age was 73,8 % were divorced, 20% never married, 32% widowed and 32% married. The most frequent diagnosis was Major Depressive Disorder of late onset followed by Personality Disorder ( 96%; 48%). The most frequent method was intoxication, (68.75% BDZ). Almost half of the attempts were highly severe. DISCUSSION: We may infer that the elderly person who attempts suicide: is female, 68-78 years old, does not have a partner, lives with someone, has Major Depressive Disorder (at least half of them had Personality Disorder) and a clinical disease. This is her first attempt, is alone at home, and she does not advise others. Reduced hearing is a very frequent co-morbidity. PMID- 12883593 TI - [Major depressive disorder in suicide attempts among over 60 years old patients]. AB - OBJECTIVE: [corrected] We analyze the presentation and duration of symptomatology and the treatment received by depressed patients prior to their suicide attempt. METHOD: This is a retrospective study using data from the medical records of patients -over 60- who were hospitalized for attempted suicide in Hospital Italiano, Buenos Aires (1999 to 2002). Diagnoses: DSM-IV criteria (by 2 trained GPs) and MMPI, Rorschach, and HDS. RESULTS: The initial symptomatology was: asthenia, weight loss, anhedonia and hopelessness. 83% with 1 to 6 months of duration. Patients who were under treatment with a psychiatrist (45.8%) were receiving: 7 adequate antidepressant doses (100% with SSRI), 3 insufficient doses (Tricyclic), and 1 no AD at all. 100% were on BDZ. 63% with neuroleptics. Of the remaining 13 patients, 11 were under treatment with a GP. This group were receiving: 2 the right antidepressant therapy (SSRI), 1 insufficient doses of Amitriptyline, and 8 were under no antidepressant therapy (72.7%). 6 of these 8 patients received BDZ and 1 was medicated with Haloperidol. CONCLUSIONS: The entire group taken into account, were not receiving the right treatment for Major Depressive Disorder. The duration of the depressive symptomatology prior to the attempt was long enough to be detected.epidemiological PMID- 12883594 TI - [Dementia, suicide attempts and suicide. A case study]. AB - The prevalence of suicide or attempts to suicide in patients with any sort of dementia has not been sharply established. The only figures found through a bibliographic search on Medline belong to a paper by Larson (13) dated 1963, who points out that only 0,8 per cent of men and 0,3 per cent of women suffering dementia commit suicide. Although persons over 65 years old are less than the 13 per cent of the world total population, they actually commit between the 17 per cent and the 25 per cent of all the suicides. Usually it was claimed that people with dementia do not commit suicide, on one hand, because of the failure in the executive abilities and in the capability to carry it on, and due to the lost of insight, on the other, both of them acting as protective factors. The aim of this article is to analyse the five clinical cases of patients with dementia that were hospitalized in a psychiatric service of a general hospital after trying to commit suicide, and then compare these cases with other four cases of patients with dementia that commit suicide reported in the international literature. PMID- 12883595 TI - [A comparative study of suicide notes in over 65 years old patients (preliminary data)]. AB - OBJECTIVE: To determine, in older suicide attempt inpatients, differences between leaving a suicide note and not leaving it as regards age, sex, marital status, religion, living alone, level of education, present psychiatric treatment, severity of suicidal attempt, previous suicidal arousal, afterwards suicidal arousal, suicidal ideation, hopelessness, depression, dementia, narcissistic personality disorder, physical pain, previous suicide attempt, familiar history of suicide or suicide attempt. MATERIAL AND METHODS: This is a comparative, prospective, observational, transversal, single blind study. RESULTS: We studied 31 patients. Statistical significant associations were found between leaving a suicide note and hopelessness (p=0.03;OR=8.57;IC=1.19-95.33) and severe suicide attempt (p=0.022;OR=8.40;IC=1.28-63.88). DISCUSSION: Although associations between writing suicide notes and hopelessness have not been reported, our results are not surprising because, on one side, the severity of the suicide attempt is more related to hopelessness than to any other depressive parameter and, on the other side, leaving a suicide note is associated to severe suicide attempts. Finally, to feel hopeless and to leave a suicide note are associated to severe suicide attempts. PMID- 12883596 TI - [Qualitative analysis of eight suicide notes of elderly people]. AB - We analize and classify eight suicide notes left by elderly patients who attempted suicide; older adults leave more frequently suicide notes. We used the typologie created by Jacobs in 1967 who classifies the notes in five categories. The issues which frequently appear are: physical illnes, cronic pain, loneliness, disability, depression and isolation. PMID- 12883597 TI - [Clinic and prevention of suicide in ederly. A literature review]. AB - In almost all countries where reliable statistics researches are conducted, the rate of suicide among elderly people rises in people over 65 years old, reaching its highest point at 85 years old. Nevertheless, it seems that this phenomena is tended to be ignored. Although the evidence given by the epidemiology, many physicians and even of psychiatrits claim that the suicide among elderly is rare. PMID- 12883598 TI - [On suicide and other essays]. PMID- 12883599 TI - Chronic renal insufficiency in pregnancy. AB - This article attempts to assess the nature, severity and management of the risks associated with pregnancy in chronic renal insufficiency and end-stage renal disease, including dialysis and transplant recipients. Women with serum creatinine levels of >125 mmol/l are at an increased risk for deterioration in renal function, hypertension with superimposed pre-eclampsia and obstetric complications. Rigid control of hypertension is crucial for a successful pregnancy outcome. A range of antihypertensive drugs are available with angiotensin converting enzyme inhibitors being contraindicated. Women on dialysis have low fertility rates that return to normal following renal transplantation. Immunosuppresive drugs are not associated with increased congenital anomalies. Transplant recipients are at an increased risk for infections that may have implications for the fetus. All groups have an increased risk for prematurity and intrauterine growth restriction. The percentage of pregnancies resulting in surviving infants in women with renal insufficiency and transplant recipients ranges from 80-100%. For women who conceive after dialysis, the likelihood of a surviving infant is approximately 50%. PMID- 12883600 TI - Elucidation of the mechanisms of hairy-cell localization in tissues and the process of the bone marrow fibrosis in hairy-cell leukemia. AB - Hairy cell leukemia is a chronic B cell leukemia with a number of distinctive features including the unusual tissue distribution of the leukemic cells, hairy cells, and the bone marrow fibrosis. We have been working, for a number of years, on the potential mechanisms behind hairy-cell localization in tissues. In this review, it is summarized how our work has shed very important information regarding these mechanisms and led, eventually, to the full elucidation of the process of the bone marrow fibrosis in hairy cell leukemia. PMID- 12883601 TI - Retinopathy of prematurity at a University Hospital in Riyadh, Saudi Arabia. AB - OBJECTIVE: To prospectively study the incidence and nature of retinopathy of prematurity (ROP) at a University Hospital in Riyadh, Kingdom of Saudi Arabia. METHODS: This study was carried out the Neonatal Intensive Care Unit of King Khalid University Hospital in Riyadh, Kingdom of Saudi Arabia. One hundred and ninety-five consecutive preterm infants with a birth weight of 2000 g or less were screened for ROP. The first examination was performed at 4-7 weeks of postnatal age. RESULTS: Mean gestational age of all premature infants was 28.4 2.4 weeks (range 22-34), mean birth weight was 1103 302 g (range 520-1960), and mean duration of oxygen therapy was 24.0 32.2 days (range 0-210). Seventy-three children developed acute ROP, giving an overall incidence of 37.4%. The incidence in preterms with birth weight of <1500 g and <1250 g was 41% and 50.7%. No infants with a birth weight of >1500 g developed ROP. Nineteen of the 73 children with ROP (26% or 9.7% of all infants studied) reached threshold ROP, and needed laser treatment or cryotherapy which induced regression in all of patients. CONCLUSION: Incidence of ROP in our patients in comparable to other reports. Screening for ROP should be carried out for all preterms of <1500 g birth weight. Such screening programs will identify those requiring retinal ablative surgery in order to induce regression of the acute ROP and prevent cicatrizing sequelae with subsequent traction retinal detachment and blindness. PMID- 12883602 TI - Pattern of infant feeding at a University Hospital in Western Saudi Arabia. AB - OBJECTIVE: To analyze the pattern of breastfeeding in mothers, identify causes of failure to breastfeed, initiate, and sustain breastfeeding practices at a University Hospital in Western Saudi Arabia. METHODS: Prospective interviews with mothers of infants seen consecutively during routine well baby clinic visits were conducted in the year October 2001 through to September 2002 using structured 18 item questionnaires. This study was carried out at King Abdul-Aziz University Hospital, Jeddah, Kingdom of Saudi Arabia. RESULTS: One hundred and twenty-eight interviews were conducted. Of the study groups 106 (83%) were breastfeeding. Of the mothers who came from low income families and no university education the figure was 90 (70%). Only 72 (56%) mothers had some form of health education regarding infant feed and most of the education came from relatives. Fourteen (10.9%) mothers reported encouragement to breastfeeding by medical personnel. Early discharge home from the newborn nursery at less than 24 hours age was significantly associated with success in establishing breastfeeding (p<0.047). Breastfeeding rates were 90% for infants in the first 6 months of life, but dropped to 72% afterwards. The reasons given for switching to formula feeding, in the order of frequency: inadequate milk supply (50%), working mothers (12.7%) and life style (10%). Mothers who were breast feeding were significantly more satisfied with their feeding practice than those whom were bottle feeding (p<0.05). CONCLUSION: The most significant factor precluding initiation of breastfeeding was late discharge from the nursery, while the most common obstacle to sustaining breastfeeding was the misconception of mothers regarding adequacy of milk supply. Intervention targeted at adoption of baby friendly nursery atmospheres and promotions of health education are economically feasible in the community. PMID- 12883603 TI - Morbidity and mortality data of cystic fibrosis patients. AB - OBJECTIVE: To identify factors that contributed to morbidity and mortality of cystic fibrosis (CF) population in the Kingdom of Saudi Arabia (KSA). METHODS: This retrospective chart review was carried out in King Faisal Specialist Hospital and Research Centre, Riyadh, KSA, during a 9 year period, November 1993 to November 2002, on confirmed CF patients, for demographic, clinical and mortality data. RESULTS: A total of 190 CF patients were diagnosed during the 9 years. One hundred and sixty-four (86%) patients are alive, 26 (14%) died. Ninety nine (52%) were males and 91 (48%) were females. Age at diagnosis 2.8 3.5 years, and period of follow up 3 3 years. In 80% of patients, symptoms started <1 year of age. Sixty-five percent of patients were in the mild to moderate malnutrition stage (<90th percentile), and 63% are in the mild to moderate stunted growth (<90th percentile). Factors that contributed to early mortality are: calculated weight/height (p-value 0.01), low albumin level at follow up (0.001), high hematocrit (HCT) (p-values=0.0002), low mean corpuscular volume (MCV) (p-0.0002), low mean corpuscular hemoglobin concentration (MCHC) (p-value 0.001), early development of antibiotic resistance (p-value=< 0.01). CONCLUSION: High HCT, low MCV, low MCHC and low albumin are factors related to poor prognosis and early death in CF patients. Iron supplement should be given to these patients even in the presence of normal hemoglobin. Early nutritional rehabilitation is needed to improve survival of our CF patients. Cohort isolation should be encouraged in CF centres. Early treatment of chronic pseudo colonization should be adopted to improve survival. PMID- 12883604 TI - Magnetic resonance imaging of normal lumbar intervertebral foraminal height. AB - OBJECTIVE: To study the pattern and changes of lumbar intervertebral foraminal heights in an asymptomatic Jordanian sample relative to age, sex, level, and correlate values with midpoint vertebral and disc heights. METHODS: One hundred and fifty-three patients (87 male and 66 female) were selected during the study period. The study was carried out at the Jordan University Hospital, Amman, Jordan from June 1999 to June 2000. Parasagittal magnetic resonance images were used to measure intervertebral foraminal heights at all lumbar levels. Values were statistically analyzed and the significance of differences in the means of foraminal heights at different levels in every age group and among age groups was determined. Foraminal height indices and correlation coefficients with midpoint vertebral and disc heights were calculated. RESULTS: The study revealed that the mean foraminal height measured is 20.9 mm 1.7 with a range of 17.1-24 mm. Foraminal heights increased significantly in a craniocaudal pattern reaching a maximum at lumber (L)2/3 in females and at L3/4 in males followed by continuous significant decrease reaching their minimum at L5/sacral (S) 1. In relation to age, foraminal heights decreased significantly in females reaching their minimum in the 7th decade. In males, foraminal heights at L3/4 until L5/S1 increased significantly reaching their maximum in the 5th decade followed by significant decrease reaching their minimum in the 7th decade. Foraminal height indices remained relatively constant. A fair degree of correlation of foraminal heights with intervertebral disc heights and vertebral body heights was evident. CONCLUSION: Foraminal heights show different level and age-dependent characteristic pattern of change between asymptomatic males and females. Changes of foraminal heights seem to directly reflect changes of vertebral body heights. These changes are considered normal age-dependent changes, and are discussed under consideration of adaptation to physical activity and changing hormonal levels. PMID- 12883605 TI - Fixed forefoot adduction after clubfoot surgery. AB - OBJECTIVE: Studying the causes of residual forefoot adduction deformity after surgical treatment of congenital clubfoot and their management. METHODS: Revision surgery was carried out by the author for 12 patients (13 feet) in King Khalid University Hospital, Riyadh, Kingdom of Saudi Arabia, during the period between January 1997 to December 1998. RESULTS: In 12 out of 13 feet, K-wire was not used to fix the navicular bone in its reduced position during primary surgery. In all cases, the navicular was found displaced during revision surgery. CONCLUSION: Revision of soft tissue surgery with relocation of the navicular bone improves the outcome of patients with residual fixed forefoot adduction after congenital talipes equinovarus surgery. PMID- 12883606 TI - Radiographic osteoarthritis and serum cholesterol. AB - OBJECTIVE: To find out the relationship between radiographic osteoarthritis (OA) of the knee, generalized OA and serum cholesterol. METHODS: Over a period of 7 months from September 1998 through to March 1999, 246 patients attending 14 primary care clinics in Northern Riyadh, Kingdom of Saudi Arabia, for different non-musculoskeletal complaints were recruited in the study. Their knees, hands and wrists were radiographed. Their ages, sex, weight, height, body mass index (BMI), fasting serum cholesterol, triglycerides and uric acid were recorded. RESULTS: There were 113 females and 133 males with average ages 46 14.2 and 51.54 16.0 years. One hundred and twenty-two (49.6%) cases of knee OA and 58 (23.6%) cases of generalized OA were found and analyzed for the association with serum cholesterol levels. Crude odds ratio (OR) for the relationship between knee OA and the third tertile of serum cholesterol was 2.33 (95% CI, 1.19-4.58) which on adjusting for age, sex, BMI, serum uric acid and triglycerides were 2.68 (95% CI, 2.00-3.64). For generalized OA, the crude odds ratio (OR) in relation to the third tertile of serum cholesterol was 1.65 (95% CI, 0.78-3.53), adjusted OR was 2.18 (95% CI, 1.55-3.15). CONCLUSION: The results showed an association between high serum cholesterol level and both knee and generalized OA. PMID- 12883607 TI - Determinants of women's fertility in Oman. AB - OBJECTIVE: International studies have the inverse relationship of women education and empowerment on fertility. Our aim is to study the association of women education, and empowerment with some of the fertility determinants in a community based survey. METHODS: A cross sectional survey of the health status of the Omani community was designed. Face to face interviews with 2037 women, who were or had been married including demographic data, fertility patterns, family planning, 2 women empowerment indices (decision making and freedom of movement), and other modules reproductive health, were carried out during the year 2000. RESULTS: About 31% of the sample was considered highly empowered in decision making and the mean number of decisions taken in the household was 4.46 for the overall sample. For freedom of movement, 29% of the sample was highly empowered with a mean number of 3.88 for the overall sample. The mean values of both indices varied significantly according to age, residence, level of education and work status. Higher freedom of movement score women were more likely to have less number of children in the first 20 years of marriage. Women of higher score of decision making index were more likely to have longer closed birth interval Those scored low in the decision making index were more likely to get a child at an earlier age. CONCLUSION: There is inverse relationship between both education and empowerment and fertility. However, the 2 indices of women empowerment; decision making and freedom of movement were not always predicting the same fertility indicator, as women's empowerment is multidimensional in nature. PMID- 12883608 TI - Results of delivery in umbilical cord prolapse. AB - OBJECTIVE: To review the peripartum clinical course of patients whose pregnancies were complicated by umbilical cord prolapse and to evaluate its impact on neonatal outcome. METHODS: All cases of cord prolapse managed in King Khalid University Hospital, Riyadh, Kingdom of Saudi Arabia between 1990-2000 were identified. There were 111 patients identified among 55,789 deliveries. Each maternal and fetal chart was reviewed for parity, age, gestational age, fetal presentation, status of membranes, time from diagnosis to delivery, mode of delivery, baby weight, Apgar scores and cord blood hydrogen ion concentration (PH). The data collected was analyzed using Gold Stat Software Package, and statistical significance was established by using analysis of variance and Chi square. RESULTS: The incidence of cord prolapse was found to be one in 503 cases (1.99 per thousand deliveries) in our study. Seventy-two (64.9%) of the fetuses were in vertex presentation and 39 (35.1%) were non-vertex, including breech and transverse presentations. Ninety one point nine percent were singletons and 8% were twins. At the time of diagnosis in 15 (13.5%) membranes were artificially ruptured and in 96 (86.5%), they were spontaneously ruptured. The cervix was fully dilated in 10% and minimally dilated in 100 (90%). Regarding mode of delivery, 7 (6.5%) were vaginal deliveries and 104 (93.5%) were cesarean sections. The interval from diagnosis to delivery ranged from 10 minutes to >20 minutes. Six (5.4%) of the babies were delivered in 10 minutes, 49 (44.1%) in 20 minutes and 56 (50.5%) in more than 20 minutes. Apgar score was less than 7 in 44 (39.6%) of the babies at one minute and in 5 (4.5%) of the babies at 5 minutes. Cord PH was less than 7 in 2 (1.8%) cases and more than 7 in 109 (98.2%). Forty one (36.9%) of the babies were admitted in neonatal intensive care unit. There was no perinatal mortality in our study group. CONCLUSION: In our review, we found that cord prolapse is not associated with higher rates of perinatal mortality or morbidity and our study supports clinical management of cord prolapse by cesarean section. The interval from diagnosis to delivery may not be the only determinant of neonatal outcome. PMID- 12883609 TI - The efficiency in the utilization of potential donors for organ transplantation in Riyadh, Saudi Arabia. AB - OBJECTIVE: Organ transplantation programs have been successful in the Kingdom of Saudi Arabia. This success is limited by organ shortage. The aim of this study is to find out the percentage of actual donors out of all potential donors in intensive care units (ICU), and to look at problems related to the donation process, particularly from the logistical point of view. METHODS: The study was conducted prospectively for a one year period, June 2001 through to May 2002, in 4 main Riyadh hospitals. Mortality data was collected by a medical professional in each ICU and analyzed on a weekly basis. Final analysis was made at the end of the year. RESULTS: Five hundred and forty-two deaths occurred in these ICUs. Fifty-four percent occurred in one hospital. The number of brain death cases in all hospitals was 114 cases. Thirty-eight cases were reported to the Saudi Center for Organ Transplantation (33%). Documentation was completed in only 23 cases (60%). In these, there was a significant delay in documentation (second test was carried out in 6-12 hours in 4 cases only). CONCLUSION: We have found that the reporting of brain death cases was low (33%). Dealing with the reported cases is inefficient since only 4 cases were able to become the actual donor out of 38 cases. We found also that there is a gross difference in the number of brain death cases among different hospitals. To improve the efficiency of ICUs in dealing with brain death cases (reporting, documentation, maintenance and consent) will require solving several problems at the medical, administrative, and religious and mass media levels. PMID- 12883610 TI - Prevalence and predictors of diabetic foot syndrome in type 2 diabetes mellitus in Jordan. AB - OBJECTIVE: To detect feet changes and to identify risk factors leading to amputation among type 2 diabetics. METHODS: A total of 1142 patients with type 2 diabetes mellitus; 595 males (52%), and 547 females (48%) were seen between January and December 2001 at the National Center for Diabetes, Endocrinology, and Genetics (NCDG) Amman, Jordan. The mean age was 56.1 years (SD=10.2) and the mean duration of diabetes was 9 years (SD=7.1). All patients had a complete medical assessment including history, physical examination, glycosylated hemoglobin (HbA1c) (the mean of the last 4 readings) and microalbuminuria. Statistical analysis were performed to identify significant risk factors leading to amputation using Epi info, version 6 software. RESULTS: Mean HbA1c was 7.4% (SD=1.4). The prevalence of hypertension was 52%, retinopathy 45% and microalbuminuria 33%. Impaired vibration, position and protective sense were found in 19%, 13%, and 18%. The prevalence of all amputations was 5%. The following were strong predictors of amputation; duration of diabetes (P= 0.04), smoking (P=0.01), microalbuminuria (P=0.02), retinopathy (P=0.008), legs hair loss (P=0.003), neurological deficit (P=0.0001), ulceration (P=0.00001) absent dorsalis pedis (P=0.0006) and insulin therapy (P=0.0001). The rate of amputation was directly proportional to high HbA1c >= 8% (P=0.01). Age and gender were not found to have an impact on prevalence of amputation. CONCLUSION: Prevalence of amputation correlates with duration of diabetes, poor glycemic control, smoking, neurological impairment, peripheral vascular disease and microalbuminuria. PMID- 12883611 TI - Knowledge, beliefs and perception of youth toward acne vulgaris. AB - OBJECTIVE: This study was carried out to assess the knowledge, concepts and perceptions of the youth towards Acne. METHODS: This cross-sectional study included 700 students of both genders from all geographic areas in Riyadh, Kingdom of Saudi Arabia during the year 2001. They were asked to complete a self administered questionnaire that contained several items measuring different areas in there beliefs, knowledge and perceptions about acne. Acne and non-acne sufferers were included. Responses were matched with demographic data. RESULTS: Five hundred and seventeen students (73.9%) responded to the questionnaire. Male and female responders were 267 (51.6%) and 250 (48.4%). More than half of them reported that they suffered from acne 276 (53.4%). Three quarters (76%) of the sample considered psychological conditions contribute to occurrence of acne, 72.1% believe that diet is one of etiologic factor. Only 15.9% reported that acne is an infectious disease. Sixty-two percent believed that acne is not a serious problem, while 56.7% consider it as both a cosmetic and health problem. CONCLUSION: The results of this study pointed out that misconceptions and false beliefs on acne are widespread and enduring among the youth. Health education program on acne is needed to improve their understanding of the condition. PMID- 12883612 TI - Predicting nodal malignancy from clinical data. AB - OBJECTIVE: A large variety of disorders can lead to lymphadenopathy. It is important and beneficial to patient management to rapidly differentiate between benign and malignant causes. The objective of the study is to identify factors predicting nodal malignancy from readily available clinical data. METHODS: A retrospective study was carried out on patients admitted to Riyadh Medical Complex, Riyadh, Kingdom of Saudi Arabia between April 1996 and March 2000 with lymphadenopathy, who underwent lymph node biopsy. RESULTS: Univariate analysis suggests 6 variables (age, sex, the presence of other physical signs, abnormal complete blood count, abnormal liver function test and negative Mantoux test) to have independent association with nodal malignancy. The multivariate logistic regression model revealed patients aged more than 40 years, males, generalized lymphadenopathy, presence of other physical signs, abnormal liver function tests and negative Mantoux test to be statistically significantly associated with nodal malignancy (p>0.05). CONCLUSION: The present logistic model can be useful in predicting nodal malignancy using routinely collected clinical data. PMID- 12883613 TI - Clinical profile of myasthenia gravis in the Sultanate of Oman. AB - OBJECTIVE: Clinical study and follow up of myasthenia gravis patients in Oman. METHODS: Follow up of 50 consecutive myasthenia gravis patients referred to the Sultan Qaboos University Hospital, Oman for a median period of 3 years from 1997 to 2000. We based the diagnosis on the clinical picture, repetitive nerve stimulation tests and edrophonium test. We performed a computerized tomography scan of the chest and anti-acetylcholine receptor antibodies. We reviewed the results of immuno modulatory treatment including thymectomy and compared these with other studies. RESULTS: Of 50 patients, 6 had purely ocular myasthenia. Of the 44 with generalized myasthenia, 28 had bulbar involvement and 12 required ventilatory support. Eight out of 29 thymectomized patients had drug free remission after 2 years. There was worsening of myasthenic symptoms in only one out of 8 pregnancies and deliveries. CONCLUSION: Bulbar and ventilatory involvement are more common in our series as compared with western data. Pregnancy and delivery were well tolerated. PMID- 12883614 TI - Pigment polyp in the common bile duct. AB - We report here a case of a young female with sickle cell disease and a past history of open cholecystectomy 10 years ago presenting with intermittent upper abdominal pain and jaundice. Abdominal ultrasonography revealed a dilated biliary tree with the possibility of a stone in the proximal common bile duct, but cholangiopancreatography showed a polypoid lesion at the junction of the common hepatic and common bile ducts. Biopsy of the lesion was benign. She underwent bile duct exploration and excision of what proved to be a pigment'polyp arising from the cystic duct stump. This is the first reported case of a pigment polyp encountered in sickle a cell disease population. PMID- 12883615 TI - Extra adrenal retroperitoneal paraganglioma. AB - We herein report a case of a 45-year-old Saudi lady not diabetic nor hypertensive who presented to the emergency room with a one day history of severe central and lower abdominal pain. On examination, she was hemodynamically stable and abdominal examination showed tenderness in the lower abdomen. Her hematological and biochemical investigations were normal. Computed tomography of the abdomen showed an 8 x 7 cm retroperitoneal mass located at the aortic bifurcation. The patient had exploratory laparotomy and complete excision of the mass. The histopathological study showed a paraganglioma. The patient had an uneventful postoperative period and follow up. PMID- 12883616 TI - Unusual presentation of echinococcal cysts. AB - We report a case of echinococcal disease affecting the brain and the kidney. The case is unusual because of multiplicity of the intracerebral lesion and sparing of the liver and lungs. The intracerebral lesion was presented with epileptic convulsions. This was successfully treated surgically. Subsequently renal echinococcal cysts were identified. These were treated by partial nephrectomy due to the location of the cysts. The patient is disease free at 8 years. PMID- 12883617 TI - Hemophilus influenzae serotype-b meningitis in an adult immunocompetent patient. AB - We present a case of meningitis caused by Hemophilus influenzae type b in an immunocompetent 41-year-old Saudi lady. The patient was successfully treated with Ceftriaxone for 10 days. A review of Hemophilus influenzae meningitis in adults and the impact of conjugated vaccine on the epidemiology of the disease are given. PMID- 12883618 TI - Retrorectal cystic hamartoma: an unusual cause of retention of urine. PMID- 12883619 TI - Angiolymphoid hyperplasia with eosinphilia presenting with postauricular swelling. PMID- 12883620 TI - Atrial septal aneurysm and stroke. PMID- 12883621 TI - Occult celiac disease in adult Omanis with unexplained iron deficiency anemia. PMID- 12883622 TI - Adhesive intestinal obstruction in infants and children. PMID- 12883623 TI - Ultrasound as a primary tool to evaluate patients with blunt abdominal trauma. PMID- 12883624 TI - SARS: The emergence of a new epidemic. PMID- 12883625 TI - End stage renal disease experience in a general hospital in Eastern Saudi Arabia. PMID- 12883626 TI - The role of magnesium sulphate in the treatment of persistent pulmonary hypertension of the newborn. PMID- 12883627 TI - Overexpression of human genes in Drosophila melanogaster by using GAL4 UAS system. AB - Many human genes determined by genomic sequencing have only few information about their functions. To fill this knowledge gap, the powerful Drosophila genetics was set as a model to elucidate human gene functions effectively. By using germline transformation together with GAL4-UAS system, we studied the possibility of expressing and functionally characterization of human genes in Drosophila. Fifty four transgenic fly lines corresponding to 10 human genes have been established. When expressed individually by crossing to an array of 6 different GAL4 driver lines, one of these genes, the translation elongation factor 1 alpha 1 (EF1 alpha 1), resulted in abnormal notum and rough eye phenotypes. This study implies the feasibility of systematically screening human gene functions by overexpression in Drosophila. PMID- 12883628 TI - Expression of recombinant human ICOS and in vitro characterization of its bioactivity on B lymphocytes. AB - Inducible costimulator (ICOS) is a novel costimulatory molecule expressed in activated T cell and has critical regulation effect on special immune response. In this study, the cDNA encoding human ICOS was cloned from activated tonsil cells via RT-PCR, and was expressed in E. coli on pET28 expression vector. The recombinant ICOS protein expressed from E. coli showed a molecular weight of 14 kD on SDS-polyacrylamide gel electrophoresis and was further confirmed by Western blot. In presence of IL-10, the purified rhICOS significantly increased in vitro B cell growth stimulated by pokeweed mitogen (PWM), and enhanced the secretion of IgG from B cells. PMID- 12883629 TI - A novel polymorphism A(+884)-->G in the hepatic lipase gene and its association with coronary artery disease. AB - Hepatic lipase (HL) activity may influence susceptibility to coronary artery disease (CAD). Association between the single nucleotide polymorphisms (SNPs) in the HL gene with the occurrence of CAD has been investigated thoroughly, but to date most studies focused on the base variation in the promoter of HL gene, little is known about the variation in the coding region. In present study, the SNP in all exons of the HL gene were analyzed. All 9 exons with their flanking sequences of the HL gene were amplified from the Chinese patients with CAD and normal controls by PCR technique, and the PCR products were detected by denaturing high performance liquid chromatography (DHPLC) and sequenced with a dideoxy terminal termination method. As the result, a novel SNP A(+884)-->G within the sixth exon of HL gene was found, the 276 codon AAA was changed into AGA and resulted in the substitution of arginine for lysine. Compared with the control group, more CAD patients carried the G+884 allele (AG+GG) (54.9% vs. 41.5%, chi(2)=6.164, df=2, P=0.046). The prevalence of the G+884 allele was significantly higher in the CAD patients than that in control subjects (31.4% vs. 21.3%, chi(2) =4.652, df=1, P=0.031). Data from the linkage disequilibrium analysis showed that the A(+884)-->G polymorphism was strong in linkage disequilibrium with the T(-2)-->C variation we identified previously(D'=0.699, 0.742 in CAD patients and controls, respectively), and the frequency of the C( 2)/G(+884) haplotype (mutation) is significantly higher in CAD patients than that in controls (0.253 vs. 0.172, P<0.05). PMID- 12883630 TI - Multi-strips on one gel method to improve the reproducibility, resolution power and high-throughput of two-dimensional electrophoresis. AB - Two-dimensional polyacrylamide gel electrophoresis is one of the most key separation tools which can reveal hundreds or even thousands of proteins at a time in proteomic research. In this paper, we report a new IPG strip application, called multi-strips on one gel (MSOG) method. By comparing the 2-DE patterns of the same sample, the different state samples and the same sample in the different second dimensional SDS running systems (large size and medium size gels), we found this new method can not only improve the reproducibility and resolution power of 2-DE pattern, but also achieve high throughput and economical format which is helpful to automatic proteomic research. PMID- 12883631 TI - Characterization of a new bradykinin-potentiating peptide (TmF) from Trimeresurus mucrosquamatus. AB - A novel bradykinin-potentiating peptide (BPP), designated as TmF, has been purified to homogeneity from the venom of Trimeresurus mucrosquamatus by 70% cold methanol extraction, Sephadex G-15 gel filtration and reverse-phase high performance liquid chromatography (RP-HPLC). The amino acid sequence of TmF was determined to be pGlu-Gly-Arg-Pro-Leu-Gly-Pro-Pro-Ile-Pro-Pro (pGlu denotes pyroglutamic acid), which shared high homology with other BPPs. The molecular mass of TmF was 1.1107 kD as determinated by electrospray ionization-mass spectrometry (ESI-MS), which was in accordance with the calculated value of 1.1106 kD. The potentiating unit of TmF to bradykinin-induced (BK-induced) contraction on the guinea-pig ileum in vitro was (1.13 +/-0.3) unit (mg/L), and TmF (5.0 x10(-4) mg/kg) increased the pressure-lowering-effect of bradykinin (5.0 x10(-5 )mg/kg) with approximate descent value of (14 +/-2) mmHg. In addition, TmF inhibited the conversion of angiotensin I to angiotensin II, 2 x10(-3) mg of TmF caused 50% inhibition (IC(50)) of angiotensin- converting enzyme (ACE) hydrolyzing activity to bradykinin. PMID- 12883632 TI - [Biological function of fusion protein ATF-PAI2CD]. AB - To express the fusion protein ATF-PAI2CD (urokinase-type plasminogen activator amino terminal fragment-plasminogen activator inhibitor type 2 with the region inter C and D helices deleted ) gene in E.coli and determine the biological characterization of fusion protein ATF-PAI2CD, the cDNA fragment encoding ATF PAI2CD was cloned into the expression vector pLY-4 and transformed into E.coli JF1125. After temperature induction, the expression amount of ATF-PAI2CD account for 15% of total bacterial protein. The result was confirmed by Western blot. ATF PAI2CD protein was isolated and purified by washing and solubilization of inclusion body, renaturation and ion exchange chromatography. The final product displayed a single band with a corresponding molecular weight 62 kD in SDS-PAGE. The purity was over 90%, the protein yield was 50% and the specific activity was 12 000 IU/mg. The PAI activity was measured by chromogenic assay. The purified fusion protein inhibited urokinase-type plasminogen activator as measured by milk agarose plate assay, and bound to human lung cancer cells via uPA receptor (uPAR), which was confirmed by radio competition experiments. The results indicate that the biological characteristics of ATF-PAI2CD were very similar to those of the wide type PAI-2 (or mutants PAI-2, PAI-2CD) and to pro-uPA in binding to uPAR-bearing cells. PMID- 12883633 TI - [Effect of calpain on the degradation of tau protein in rat brain cortex extracts]. AB - Calpain is a calcium-activated protease and has two ubiquitously distributed mammalian isoforms, namely calpain 1 (calpain I, mu-calpain and CAPN1) and calpain 2 (calpain II, m-calpain and CAPN2). Calpains regulate the function of many proteins by limited proteolysis. To determine the nature of different subtypes of calpain on degradation of microtubule-associated protein tau, the rat cortex extracts were incubated with 0.2 mmol/L, 1 mmol/L, 3 mmol/L and 5 mmol/L of CaCl(2 )for 15 min at 37 degrees C, respectively, and it was found that Ca(2+) treatment at concentrations 1-5 mmol/L led to significant proteolysis of the tau protein and this degradation was blocked by calpain inhibitor, calpeptin. In addition, when the extracts containing 1 mmol/L CaCl(2 )were treated with mu calpain inhibitor (0.05 micromol/L of calpastatin) or m-calpain inhibitor (100 micromol/L calpain inhibitor IV) or both, the Ca(2+)-induced degradation of tau protein was blocked to about 8.6% 92.5% and 97.8% compared with the group with 1 mmol/L CaCl(2), respectively. These data suggest that both mu-calpain and m calpain in brain cortex extracts are activated by Ca(2+) and both of them degraded tau protein, although, m-calpain plays a more important role in proteolysis of the tau protein. PMID- 12883634 TI - [Identification of protein spots in silver-stained two-dimensional gels based on chemically assisted fragmentation by MALDI-TOF-PSD mass spectrometry]. AB - Guanidination of the epsilon-amino group of lysine-terminated tryptic peptides,accomplished with O-methylisourea hydrogen sulfate, converted lysine into homoarginine residues, improving detection in MALDI-MS. Then tryptic peptides were labeled with sulfonic acid groups at the N-termini using 3 sulfopropionic acid NHS-ester chemistry. The derivatives enhanced post-source decay (PSD) fragmentation signals and produced a spectrum containing only y-ions. This facilitated de novo peptide sequencing, so it is an important contribution to unambiguous protein identification in proteome research. This method was successfully applied in nasopharyngeal carcinoma(NPC) proteome study. PMID- 12883635 TI - [Mapping of FHL2 transcription activation domain]. AB - FHL2, a member of LIM-only protein family, plays an important role in transcription regulation, apoptosis, cancer development and progression. In this study, a mammalian transcription activation system was constructed by using DNA binding domain(DBD) of GAL4 and luciferase reporter gene with DBD binding sequence, and used for mapping of FHL2 transcription activation domain. First, the coding sequence of GAL4-DBD was inserted into expression vector pcDNA3, generating the pDBD recombinant plasmid, then the wild-type FHL2 and its mutants were fused in-frame with GAL4-DBD, resulting in expression vectors for wild-type FHL2 and its mutants. All of the recombinant plasmids were transfected into 293T cells. Western blot assay showed that all of the fusion proteins were expressed. The analysis of FHL2 transcription activation properties by using the GAL4 luciferase reporter gene indicated that wild-type FHL2 had activation activity in both 293T and MCF-7 cells. The deletion of the half LIM domain at the N-terminus severely impaired the capacity of FHL2 to stimulate transcription. The mutant lacking the LIM domain at the C-terminus was totally inactive, while the deletion of two LIM domains at the C-terminus partially recovered its ability to stimulate transcription. The deletion of the second LIM domain at C-terminus did not alter the activation capacity of FHL2. These results suggest that the last LIM domain at the C-terminus of FHL2 is critical for its transcription activation function, the second LIM domain at the C-terminus may be a negative regulation region, but this negative regulation depends on the last LIM domain. Mapping of transcription activation domain of FHL2 lays solid basis for further study on various FHL2 functions. PMID- 12883636 TI - [Cloning, expression, and antibody production of mouse ISP2]. AB - A cDNA encoding mouse implantation serine proteinase 2 ISP2 was amplified from total cDNAs of mouse uterus implantation sites on D4.5 of pregnancy by PCR, and sequenced GenBank accession No. A442918 . DNA sequencing indicated that the ISP2 cDNA had an unreported 204 bp sequence at 3' untranslated region besides the open reading frame encoding 279 amino acid residues, which was identical with literature. In order to obtain recombinant ISP2 rISP2 an expression plasmid pGEX-4T-2/ISP2 was constructed and transformed into E.coli BL21(DE3) strain. Expressed fusion protein GST-ISP2 was purified by SDS-PAGE and digested with thrombin, and the digestion mixture was subjected to SDS-PAGE again to recover rISP2. Rabbits were immunized using rISP2 as immunogen, and the polyclonal anti ISP2 antisera were obtained. Immunohistochemical analysis using this antisera showed specific and high expression of ISP2 in mouse endometrial gland epithelium in early pregnancy. PMID- 12883637 TI - Photodynamic effects of 5-aminolevulinic acid and its hexylester on several cell lines. AB - 5-aminolevulinic acid (ALA) and its hexyl-ester (He-ALA) has shown promising results in photodynamic detection and therapy of tumors. In this work, the photodynamic effects of ALA and He-ALA on neuroblastoma cells, hepatoma cells and fibroblast cells were comparatively studied. With the detection of fluorescence emission spectra, protoporphyrin IX (PpIX) induced by ALA or He-ALA was observed in these three cell lines. Confocal laser scanning microscope showed the diffuse PpIX fluorescence in cytoplasm of neuroblastoma cells. The kinetics of PpIX accumulation were different in these three kinds of cells. The PpIX content in hepatoma cells and fibroblast cells continuously increased with the incubation time of drugs until 12 h, while in neuroblastoma cells the PpIX content saturated around 8 h after incubation with ALA or He-ALA. In addition, the PpIX concentration in neuroblastoma cells was obviously higher than that in hepatoma cells and fibroblast cells, indicating that the PpIX production is cell line dependent. When incubated with ALA and irradiated with light, near 90% neuroblastoma cells were destroyed, while for hepatoma cells and fibroblast cells the death rate was around 50%. The results demonstrate that neuroblastoma cells are more sensitive to ALA-PDT and the neuro-tumor cells may be well suited for the treatment of ALA mediated photosensitization. Comparing to ALA, He-ALA can reach the similar results concerned PpIX production and PDT damaging in all three kinds of cells but with 10 times lower incubation concentration, demonstrating that He-ALA has higher efficiency than ALA on inactivation of cancer cells in vitro. PMID- 12883638 TI - Identification of interaction between PAI-2 and IRF-3. AB - HeLa cells transfected with plasminogen activator inhibitor-2 ( PAI-2 ) were protected from TNF- alpha-induced apoptosis. The apoptosis protection by PAI-2 is dependent on a 33 amino acids fragment between helix C and D of PAI-2, which may be due to the interaction of PAI-2 with some intracellular proteins. In this study, the yeast two-hybrid system was used to screen a HeLa cells cDNA library constructed during apoptosis with the fragment between helix C and D of PAI-2 as bait. We retrieved a clone that encodes 98 amino acids of C-terminus of interferon regulatory factor-3 (IRF-3). Co-immunoprecipitation experiments confirmed the interaction between PAI-2 and IRF-3 in vivo. IRF-3 belongs to a family of the IRF transcription factors and has been shown to participate in a large number of biological processes. These data suggest that IRF-3 may be involved in the apoptosis protection and antiviral function of PAI-2. PMID- 12883639 TI - Expression, purification, and characterization of recombinant Saccharomyces cerevisiae adenosine kinase. AB - Adenosine kinase (AK), a key enzyme in the regulation of the cellular concentrations of adenosine (A), is an important physiological effector of many cells and tissues. In this article, we reported that ak, which encoded adenosine kinase, was cloned from Saccharomyces cerevisiae, sequenced, and overexpressed in E. coli using the pET16b expression system, and the recombinant protein was purified to apparent homogeneity using conventional protein purification techniques. Kinetic analysis of S. cerevisiae AK revealed K(m) values of (3.5+/ 0.2) micromol/L for adenosine and (100.0+/-11.0) micromol/L for ATP, with k(cat) of (1530+/-20) min(-1) for adenosine and (1448+/-25) min(-1) for ATP. The determination of the K(m) value for other nucleosides and deoxynucleoside indicated that the nucleoside specificity of this enzyme from yeast was quite high. PMID- 12883640 TI - [Use of three-dimensional fluorescence deconvolution microscopy for study of spatial distribution of secretory vesicles in living cells]. AB - A three-dimensional image of a living cell is helpful for cell secretion study. In this report, the three-dimensional fluorescence deconvolution microscopy for observing living cells was studied, because this technique can obtain a quick three-dimensional imaging with minimal fluorescence quenching and cytotoxicity for living cell observation. The property of three-dimensional point spread function (PSF) of imaging system was analyzed. The relationship between experimental and theoretical PSF was illustrated, and the theoretical PSF was proved that it could reflect the principle of imaging system with NA 1.65 objective in use. Three-dimensional deconvolution algorithm in this report was proved effective by well-defined three-dimensional specimens. Furthermore, the rat pancreatic beta cell secretory vesicles labeled by acridine orange was observed by using this technique. Results showed that the blurring induced by out of-focus light was removed by the deconvolution algorithm effectively, under current experiment conditions (with NA 1.65 objective) the experimental PSF approximated the theoretical PSF very well, and deconvolved living cell images exhibited the spatial distribution of the secretory vesicles clearly. PMID- 12883641 TI - [Thermodynamic and kinetic analysis of unfolding of P23k protein isolated from spinach photosystem II]. AB - The unfolding of 23kD (P23k) protein isolated from spinach photosystem II particle was studied by high pressure and fluorescence spectroscopy. The thermal equilibrium study indicated that the protein could be totally unfolded by 180 or 160 MPa at 20 degrees C and 3 degrees C, respectively. The standard free energy and standard volume change of the protein for unfolding at 20 degrees C is 23.45 kJ/mol and -150.3 ml/mol, respectively. Kinetics study indicated that at 20 degrees C the activation volume for unfolding, delta V(u)(++), was negative ( 66.2 ml/mol), meanwhile the activation volume for folding, deltaV(f)(++), was positive (84.1 ml/mol). The rate constants for folding and unfolding (K(0f), K(0u)) were 1.87 s(-1) and 1.3x10(-4) s(-1), respectively, these results provide some clues to explain why the protein is so sensitive to pressure. PMID- 12883642 TI - Ghrelin: a recently discovered gut-brain peptide (review). AB - In the 70s, several new, both peptidyl and non-peptidyl, derivatives that stimulate and amplify pulsatile growth hormone (GH) secretion, independently from growth hormone releasing hormone (GHRH), were synthesized. The family of these molecules have been named growth hormone secretagogues (GHSs). The subsequent discovery of the natural receptor for GHSs (GHS-R) suggested existence of a new regulatory system, participating in GH secretion control. Three years later a natural ligand for GHS-R was identified and was designated 'ghrelin'. Subsequently, it has been found that ghrelin exerts pleiotropic activity. It influences not only GH release but also food intake, function of gastrointestinal tract and cardiovascular system, sleep patterns as well as cancer proliferation. The current knowledge on ghrelin, its structure and function, is reviewed in this article. PMID- 12883643 TI - Secretory immune system in human intrauterine development: immunopathomorphological analysis of the role of secretory component (pIgR/SC) in immunoglobulin transport (review). AB - Several proteins, such as polymeric immunoglobulin (Ig) receptor/secretory component (pIgR/SC), immunoglobulins (Igs) and joining (J) chain, and cellular components of the secretory immune system (SIS) of the human embryo and fetus were analyzed and compared with reviewed information concerning SIS organization and function. All organs and structures, including extracorporeal ones, of 18 embryos (4-8 weeks of development) and 45 fetuses (9-38 weeks) were studied using methods of pathomorphology, immunohistochemistry and morphometry. This approach enabled us to analyze the problem in the whole organism during its entire development. SC, Igs and J chain were already widely distributed in 4-week-old embryos and were later seen in the mucosa and glands of the digestive, respiratory and urogenital tracts, ovaries, testis, endocrine glands, extracorporeal tissues, blood-brain and other blood-organ barrier structures, as well as in serous membranes. The presence of protein transport and later of cellular components suggests an active role for SIS not only in mucous membranes, but also in blood-tissue barriers. Loss of morphological contact between epithelial structures and mucous membranes during organogenesis of some endocrine organs (hypophysis, pancreatic islets, etc.) is followed by the disappearance of SC as a result of cessation of Ig exocytosis. Secretion of Igs increased in the epithelium and glands of the digestive and respiratory tracts following massive antigenic attack in cases of acute chorioamnionitis. All of this demonstrates that SIS is a widely branching immune system which appears and acts early in the human embryo, before the lymphoid system is formed, using IgG and IgA of maternal origin. IgA and IgM-synthesized lymphocytes appear in 9-week-old fetuses. PMID- 12883644 TI - Combination of nucleic acid and protein isolation with tissue array construction: using defined histologic regions in single frozen tissue blocks for multiple research purposes. AB - Precise dissection of defined histological regions for nucleic acid and protein isolation is a precedent step in finding out cancer-related alterations, and high quality tissue microarrays are demanded in the validation of screened genetic alterations by multiple in situ approaches. In this study, a combined technique was developed by which sample isolation and tissue array construction could be performed on the defined morphological region(s) in single tissue block. The RNA and protein samples generated from the selected portions were of good quality and sufficient for multiple experimental purposes. The frozen tissue arrays constructed on a novel recipient are suitable for multiple in situ evaluations including immunohistochemical staining and mRNA hybridisation. In most cases, the data obtained from in situ assays coincided well with the ones revealed by RT-PCR and Western blot hybridisation. The potential experimental bias caused by cell contamination can be amended by tissue array-based retrospective examination. The combination of tissue-selective sample preparations with tissue array construction thus provide a tool by which comprehensive cancer research can be performed on defined histological regions in a series of single frozen tissue blocks. PMID- 12883645 TI - IL-10 in vivo gene expression in a cell-induced animal model of proliferative vitreoretinopathy. AB - Due to inhibitory activities on cell-mediated immune responses, interleukin-10 (IL-10) has been proposed as a good candidate to treat inflammatory eye disease and proliferative vitreoretinopathy (PVR). In this study we evaluate the effect of human IL-10 (hIL-10) expression in a cell-induced animal model of PVR. Rabbit dermal fibroblasts were genetically modified by infection with retroviral particles carrying the neomycin resistance gene (neoR) alone or in combination with the hIL-10 gene. PVR was induced in rabbits by intravitreal injections of RDF hIL-10 or RDF neo. Some rabbits received instead injections of soluble recombinant hIL-10 (rhIL-10). PVR was graded by fundoscopy. Eyes were enucleated for histology at day 28. ELISA was performed to measure hIL-10 production in RDF supernatants and in vitreous samples, 24 h after injection. Results showed that in vitro hIL-10 production by RDF was 24,500 pg/10(6) cells/ml/24 h. In vivo IL 10 secretion was detected in all rabbits injected with RDF hIL-10 but was undetectable in control rabbits. Similar clinical grades of PVR were found in rabbits injected with RDF hIL-10 or RDF neo. Histology showed that all eyes injected with RDF hIL-10 had significant inflammatory infiltration whereas only one control eye was clearly inflamed. Rabbits injected with soluble rhIL-10 had normal fundoscopy and normal histology. In conclusion, our results show that in vivo, in a cell-induced model of PVR, hIL-10 has no effect in the clinical progression of PVR. Histology, however, shows that pro-inflammatory effects seem to overcome its suppressive properties. PMID- 12883646 TI - Procaspase-3 and its active large subunit localized in both cytoplasm and nucleus are activated following application of apoptotic stimulus in Ramos-Burkitt lymphoma B cells. AB - Ramos-Burkitt lymphoma (Ramos-BL) B cell line is a neoplastic model of normal B cell selection by apoptosis at the germinal center site during maturation of the humoral immune response and can be triggered into apoptosis by cross-linking their surface antigen receptor with antibodies directed against immunoglobulin (Ig)M (anti-IgM) or by treating with the calcium ionophore ionomycin. We have recently demonstrated that anti-IgM and ionomycin trigger significant activation of caspase-3, -7 and -8 and for cleavage of the resident nuclear proteins poly(ADP-ribose) polymerase (PARP) and lamin B1 in Ramos-BL B cells, suggesting that these caspases may be localized to the nucleus as well as to the cytoplasm of Ramos-BL B cells. In order to examine this hypothesis further, we fractionated Ramos-BL B cells into their cytosolic and nuclear components and examined for expression of the endogenous proform and active large subunit of caspase-3; procaspase-3 and its active p17 large subunit were identified in both the cytosolic and nuclear fractions of Ramos-BL B cells. Immunofluorescence staining together with ordinary and confocal microscopy confirmed the observations that procaspase-3 immunoreactivity was clearly identified in the cytoplasm and nucleus while Fas ligand staining was localized to the cell surface and PARP immunoactivity to the nucleus, which were used as controls; procaspase-3 exhibited granular nuclear immunoreactivity whereas PARP displayed diffuse nuclear immunoreactivity; both of which was more intense in the internucleolar regions. Taken together, we now present evidence that procaspases and their active large subunits are found in both the cytoplasm and the nucleus and that procaspases localized not only in the cytoplasm but also in the nucleus are activated following application of apoptotic stimulus in Ramos-BL B cells. PMID- 12883647 TI - Common genes responsible for differentiation and senescence of human mucosal and epidermal keratinocytes. AB - Serial subcultures of normal human oral keratinocytes (NHOKs) and normal human epidermal keratinocytes (NHEKs) to the postmitotic stage result in terminal differentiation and replicative senescence. In order to investigate the common mode of differentiation and/or senescence between mucosal and epidermal keratinocytes, gene expression profiling on both NHOKs and NHEKs was performed by a cDNA microarray analysis. Primary NHOKs and NHEKs were serially subcultured, and the expression level of 3,063 genes was compared between the exponentially growing and senescent cultures. The senescent NHOKs and NHEKs highly expressed 55 and 37 genes, respectively. Among these genes, 16 genes were common in both NHOKs and NHEKs while the other genes were upregulated either in the NHOKs or in the NHEKs. Furthermore, the expression levels of the common genes did not change in the human diploid fibroblasts during the subcultures. These results suggest that subculture-induced differentiation and/or replicative senescence in NHOKs and NHEKs has similar characteristics, but that the pathways leading to these processes are distinct and keratinocyte specific. PMID- 12883648 TI - Intra-plaque production of platelet-activating factor correlates with neoangiogenesis in human carotid atherosclerotic lesions. AB - Platelet-activating factor (PAF) is a phospholipid mediator synthesized by activated inflammatory and endothelial cells. Recently PAF has been shown to contribute to neoangiogenesis in several experimental models. Here we evaluated the presence of PAF and its potential role in neovascularization within human atherosclerotic plaques. The amount of PAF extracted from 18 carotid plaques (266.65+/-40.07 pg/100 mg dry tissue; mean +/- SE) was significantly higher than that extracted from 18 normal arterial specimens (6 from carotid artery and 12 from aorta) (4.72+/-2.31 pg/100 mg dry tissue; mean +/- SE). The levels of PAF significantly correlated with the infiltration of CD68-positive monocytes and the extent of neovascularization, detected as von Willebrand Factor-positive cells. The amount of PAF also correlated with the area occupied by TNF-alpha-expressing cells. The absence of enhanced level of PAF in the circulation of atherosclerotic patients suggests a local production of this mediator within the plaque. The lipid extracts of atherosclerotic plaques containing high levels of PAF bioactivity, but not those of control arteries, were angiogenic in a murine Matrigel model. WEB 2170, a specific PAF receptor antagonist, significantly prevented angiogenesis induced by the lipid extracts of atherosclerotic plaques. Our results indicate a local production of PAF within the atherosclerotic plaques and suggest that it may contribute to intra-plaque neoangiogenesis. PMID- 12883649 TI - Malignant melanoma associates with deficient IFN-induced STAT 1 phosphorylation. AB - STAT 1, a member of signal transducers and transcription activators of STAT family proteins, has been implicated as important mediator of IFN signaling. Functional activation of STAT 1 requires tyrosine and serine phosphorylation. Defects in its expression or activation in response to IFNs were observed in numerous pathological conditions including cancer. To further explore cancer associated impaired STAT 1 response to IFNs, the inducibility of serine (S 727) and tyrosine (Y 701) phosphorylation by IFN-alpha/-gamma was assessed in 21 melanoma cell lines and in 35 primary cultures derived from melanoma patients. STAT 1 levels and inducibility of its activated phospho-forms were detected by Western analysis using specific polyclonal and monoclonal antibodies. All cell lines as well as patient melanoma samples expressed STAT 1 with variable signal intensity. Significant impaired IFN-induced STAT 1 S 727 phosphorylation was observed in both model systems with average of 77% of non-responders recorded in patient melanoma cells and 76% in melanoma cell lines. Failure of PY 701 induction occurred in patient samples (63% after IFN-alpha and 34% after IFN gamma induction) and to a lesser degree in cell lines (i.e. response absence to IFN-alpha in 5 and to IFN-gamma in 2 melanoma lines). Our study demonstrates STAT 1 functional abnormalities in melanoma cells. On the basis of detailed analyses of patient melanoma cells with respect to the inducibility of STAT 1 phosphorylation by IFNs, four categories of patients could be distinguished: a) activation on both S 727 and Y 701, b) not inducible response, c) activation on Y 701 but not on S 727, d) heterogeneous response. Clinical study is now in progress to establish the significance of in vitro STAT 1 activation for predicting the response to IFN-based therapy and to explore biological consequences in cases responding in vitro to IFN-induced STAT 1 activation on only one of the critical amino acid residues. PMID- 12883650 TI - Ezrin, actin and cytoskeleton in apoptosis of esophageal epithelial cells induced by arsenic trioxide. AB - Ezrin is a key protein in membrane-cytoskeleton interaction. Expression of ezrin in actin-rich cell surfaces may play a role in the modulation of cell shape and adhesion. The aim of this study was to detect ezrin, actin and cytoskeleton and to explore their relationship in the apoptosis of esophageal epithelial cells (SHEE) induced by arsenic trioxide (As2O3). The SHEE is an immortalized human fetal esophageal epithelial cell line, and the cells were treated by administering 5, 10 and 20 micromol of As2O3. the proliferation and apoptosis of SHEE cells were examined by flow cytometry with propidium iodide staining. Ezrin expression was detected by immunocytochemistry and Western blotting. Actin filament was stained by FITC-labeled phalloidin and detected quantitatively by fluorescent microscopy. Cell morphology and microfilaments were examined by electron microscopy. The results revealed that As2O3 induced an inhibition of proliferation and the promotion of apoptosis in SHEE cells. The ezrin, actin and cytoskeleton were decreased after As2O3 treatment and the cellular morphology of apoptosis developed. Our results suggested that the morphological changes of arsenic-induced apoptosis of human esophageal epithelial cells were initiated by ezrin and actin-cytoskeletal aberrance. PMID- 12883651 TI - The significance of Elf-1 expression in epithelial ovarian carcinoma. AB - Elf-1 is a member of the Ets transcription factor family that regulates the genes involved in cellular growth and differentiation. Enhanced expression of Elf-1 has been reported in prostate cancer, breast cancer, and osteosarcoma. The aim of this study was to elucidate the involvement of Elf-1 in ovarian carcinogenesis. We analyzed serial frozen tissue sections from 26 patients with epithelial ovarian carcinoma and 10 patients with benign cystadenoma of ovary for Elf-1 protein expression, using fluorescent immunohistochemistry. We analyzed the relationship between the percentages of Elf-1-stained cells and patient characteristics, including histological classification, clinical stage, histological grade, clinical outcome, and survival rate. Elf-1 was weakly detected in some benign cystadenomas (0-5.5%). There was, however, abundant Elf-1 immunoreactivity in the nucleus of the ovarian carcinoma cells along with a little cytoplasmic staining. Scoring on the basis of the percentage of nuclear positive cells indicated that nuclear Elf-1 expression was significantly associated with PCNA-labeling index (p=0.04), clinical stage (p<0.01), histological grade (p<0.01), and clinical outcome (p=0.02). However, there was no relationship between Elf-1 expression and histological classification. Survival data were available for all patients and demonstrated that Elf-1 expression was significantly associated with poor prognosis (p<0.01). Our results demonstrate that Elf-1 expression in ovarian carcinoma correlates with the malignant potential of this tumor. PMID- 12883652 TI - Alternative splicing of the human estrogen receptor alpha primary transcript: mechanisms of exon skipping. AB - The 1785 nucleotides of the coding region of the estrogen receptor alpha (ER alpha) are dispersed over a region of more than 300,000 nucleotides in the primary transcript. Splicing of this precursor RNA frequently leads to variants lacking one or more exons that have been associated to breast cancer progression. The most frequent splice variant lacks exon 4 and is expressed in the human mammary carcinoma cell line MCF-7 at a level similar to that of the full-length messenger. The in silico analysis of ER-alpha splice sites by Hamming clustering, a self learning method trained on more than 28,000 experimentally proved splice sites, reveals high relevance for the 5' and 3' splice sites of exon 4. The splicing analysis of transfected mini-gene constructs containing drastically shortened introns excludes that weak splice sites, intron or exon lengths or splice enhancers are responsible for exon skipping. Exon 6 is never skipped in MCF-7 cells but is spliced out from mini-gene derived primary transcripts if inserted between exons 3 and 5 instead of exon 4. As a consequence, it appears that a particular splice site affinity of exon 3 donor (5' splice site) and exon 5 acceptor sites (3' splice site) is responsible for skipping of the exon in between. PMID- 12883653 TI - Effect of CaCl2 concentration on the rate of foreign gene transfer and expression by in vivo electroporation in the mouse ovary. AB - We tested the effect of electroporation (EP) medium composition on the rate of gene transfer and expression in the mouse ovary in vivo. FITC labeled oligonucleotides were dissolved in a medium with varying levels of CaCl2 concentration from 0 to 250 mM, and transferred by in vivo EP. Gene transfer efficiency was assessed by examining fluorescence signal intensity with a fluorescent microscope at 3 h after in vivo EP. The results indicated that CaCl2 concentration at 50 mM gave the highest transfer efficiency of the oligonucleotides only in the presence of phosphate buffered saline (PBS). Without PBS or CaCl2, the oligonucleotide transfer was negligible. A further increase in CaCl2 from 50 to 250 mM lowered the transfer efficiency. Little fluorescence signal was attained by substituting CaCl2 for MgCl2, NaCl or KCl. Addition of glycerol to the EP medium with 50 mM CaCl2 did not improve the transfer efficiency in the presence of PBS, although a marginal increase was observed in the absence of PBS. The stimulating effect of increased CaCl2 concentration from 0 to 50 mM was further evaluated by examining the intensity of reporter protein expression after transferring the bacterial lacZ gene. The results of X-gal staining demonstrated that CaCl2 with a range of 20 to 100 mM, showed enhanced gene expression in comparison with 0 mM. However, no remarkable difference was observed between the different CaCl2 concentrations, suggesting that the stimulating effect of CaCl2 on gene transfer and expression in the mouse ovary in vivo may not necessarily parallel in terms of the optimal concentration. PMID- 12883654 TI - Parallel comparison of chronological risk changes among cancers of the breast, the uterine cervix and the stomach, as tested in 9 female populations of the world from early 1960s to mid 1980s: a stochastic study. AB - The purpose of this study was to investigate comparatively the chronological changes of age-adjusted incidence rate (AAIR) as well as age-specific incidence rate (ASIR) of cancers of the breast (Br), the uterine cervix (Cer) and the stomach (St) from early 1960s to mid 1980s (5 data collections) using the data sets of 9 female populations world wide, the total data set number amounting to 45 per tumor. More specifically, we wanted to see whether or not there was any regular relationship between the differential time trends of the above 3 human neoplasias and their oncogene-tumor suppressor gene balances. Our investigation proceeded in 3 steps as follows: step 1, straight line regression analysis of log AAIR was applied to each of tumor pairs Br(x)-Cer(y), Br(x)-St(y) and Cer(x) St(y). Step 2, direct successive elimination test (Gauss) of log AAIR was applied to each of 6 tumor pairs (Br-St, Br-Cer, Cer-St, Cer-Br, St-Br and St-Cer) to assess the fitness of a test tumor (x-partaner of fitness test) to either the oncogene type equilibrium model (r=-1.000) or the tumor suppressor gene type equilibrium model (r=+1.000). For each of 6 tumor pairs, the fitness test was repeated 3 times using i) the original data set in the ordinary (x,y) framework, ii) the rect-type data sets in the rect-(x,y) framework, and iii) the para-type data set in the para-(x,y) framework. The fitness of a test tumor (x partner in the calculation) to an equilibrium system was assessed in terms of the correlation coefficient value r within the range of -1.000 to +1.000 (step 3). Staging of the whole carcinogenesis process was attempted for each of 3 human neoplasias using either the ASIR profiles of a high- and low-risk populations, and/or those of early 1960s and mid 1980s. Results of key importance are given as follows: i) Br with rapidly growing cancer risk, Cer with rapidly declining cancer risk and St with slowly declining cancer risk were clearly distinguished from one another by the single regression analysis. ii) The fitness test demonstrated that Br, Cer and St each were found to have one score in the positivity test of oncogene activation (r=-1.000), whereas Br with 4 scores, Cer with 2 scores and St with 3 scores of tumor suppressor gene inactivation (r=+1.000) were found to have differential openness to the fitness test of Gauss for the detection of tumor suppressor gene inactivation. It is indicated that the differential opennesses of a window to tumor suppressor gene inactivation observed in 3 human neoplasias represent a measure of the time-linked aggressiveness of cancer risk. iii) The comparison of 4 ASIR profiles of Br suggested the presence of 3 stages of carcinogenesis progression, of which the 1st stage was tentatively classified as of heredity-dependent origin, and the 2nd and the 3rd stages were classified as of environment-dependent origin. The St ASIR profile represents a fusion product of very minor heredity-dependent stage I and a major environment-dependent stage II. In contrast, the Cer ASIR profile completely lacked a heredity-dependent stage. The integrity of the above findings is discussed in the light of a number of pioneering achievements along the line of the steroid criminal theory of carcinogenesis in humans and in non-human mammals. PMID- 12883655 TI - Acute intracerebroventricular administration of either carboxyl-terminal or amino terminal fragments of agouti-related peptide produces a long-term decrease in energy expenditure in rats. AB - We investigated if agouti-related peptide (AgRP), an endogenous antagonist of melanocortin receptors (MC3-R and MC4-R), effects energy expenditure in rats. Fragments of the carboxyl-terminal, AgRP (83-132), and the amino-terminals, AgRP (25-51) and AgRP (54-82), were administered intracerebroventricularly (ICV). Food intake, body weight and fat weight changes were measured 5 and/or 24 h after a single ICV injection of the fragments. Oxygen consumption and colonic temperature were measured as indices of energy expenditure, during 3 and 24 h after the ICV injections, respectively. An oral glucose tolerance test was performed 24 h after ICV AgRP (83-132) injection. Binding experiments were performed in HEK-293 cells that over-expressed human MC4-R. AgRP (83-132), but not AgRP (25-51) nor AgRP (54 82), induced a potent and long-lasting increase in the cumulative food intake. Both the carboxyl-terminal and amino-terminal AgRP fragments significantly decreased oxygen consumption and colonic temperature. Despite the absence of hyperphagia and cross-reactivities with MC4-R, AgRP (25-51) and AgRP (54-82) significantly increased body weight and epididymal/mesenteric fat weight. AgRP (83-132) did not affect glucose and insulin responses to the oral glucose tolerance test. AgRP causes a potent and long-lasting decrease in energy expenditure; an effect that is exhibited by carboxyl-terminal fragments and amino terminal fragments that lack antagonist activity at the MC receptors. This suggests that the amino-terminal region of AgRP plays a regulatory role in energy metabolism. PMID- 12883656 TI - The influence of expanded unmethylated alleles for FRAXA/FRAXE loci in the intellectual performance among Brazilian mentally impaired males. AB - Both fragile X (FRAXA) syndrome and fragile XE (FRAXE) disorder are caused by an expansion of a polymorphic trinucleotide repeat and are associated with mental impairment. Based on the size and methylation status of the expansion, individuals are classified as having normal, intermediate, premutation or full mutation alleles. Unlike individuals with full mutations, carriers of intermediate and premutated alleles should not exhibit obvious clinical symptoms, since the FMR1 (FRAXA) or FMR2 (FRAXE) genes are not transcriptionally silenced. However, there are data suggesting a phenotype consequence of the FRAXA premutation alleles. We have investigated a population consisted of 276 males with idiopathic mental retardation or learning disability and a control sample of 207 non-affected boys in order to determine if there was a possible phenotype consequence of the expanded unmethylated alleles for FRAXA/FRAXE loci. Direct molecular diagnosis for the FRAXA/FRAXE loci were performed in both populations by using PCR technique and sizing of the amplification products by electrophoresis in denaturing polyacrilamide gels. No FRAXA/FRAXE premutations or FRAXE mutated alleles were observed. The 25 FRAXA full mutations alleles detected were confirmed by Southern blot analysis. We found an excess of intermediate alleles for both FRAXA and FRAXE in the target population, but it did not reach statistically significant difference. This suggests that relatively large unmethylated repeats may not be associated with an abnormal cognitive and/or behavioral phenotype. However, recent evidence that FRAXA premutation alleles in males have increased FMR1 message levels emphasizes the need of more studies using large sample sizes and proper control population to resolve this contradictory observation. PMID- 12883657 TI - A bioinformatics-based approach for the prediction and identification of novel proteins potentially involved in phosphorylation signalling pathways. AB - Together with the explosion in the availability of genome data of a number of organisms including human and mouse, various methods and programs for computational prediction of protein-coding genes and annotation of functional proteins have dramatically increased. For the last decade there has been intense interest in the role of protein phosphorylation which is involved in post translation modification mechanisms critically regulating inter/intracellular communication, patho/physiological responses and homeostasis during many biological processes. In the present study a total of 202 functionally uncharacterized human full-coding cDNA sequences were investigated using a bioinformatics-based approach. Ten novel potential substrates for protein kinases have been identified which may play multiple roles in regulating intracellular phosphorylation signalling pathways. In addition, 5 of those may be involved in the human-only post-translation mechanism regulated by specific protein kinases. The data presented here therefore would greatly contribute toward the understanding of human molecular basis and cellular signalling networks. PMID- 12883658 TI - Identification and characterization of human ZPBP-like gene in silico. AB - PP1R1B-ERBB2-GRB7 locus on human chromo-some 17q12 is frequently amplified in gastric and breast cancer. Because recombination hot spot or fragile site is located around the terminus of amplified region (amplicon), we searched for a novel gene closely linked to the teromeric end of the ERBB2 amplicon. Here, we identified and characterized the ZPBP-like (ZPBPL) gene by using bioinformatics. ZPBPL gene, corresponding to BC043152 cDNA, was found to consist of seven exons. ZPBPL (316 aa) and ZPBP (351 aa) proteins, showing 34.8% total amino-acid identity, shared the zona pellucida binding protein homologous (ZPBH) domain with conserved 15 cysteine residues. ZPBPL was a secreted-type glycoprotein with the ZPBH domain, while ZPBP was a type 2 transmembrane protein with the extracellular ZPBH domain. ZPBPL mRNA was co-expressed with ZPBP mRNA in testis, germ cell tumor, and brain medulla. ZPBPL might be implicated in the gamete interaction during fertilization just like ZPBP. The MGC9753-ERBB2-MGC14832-GRB7-ZNFN1A3 ZPBPL-PRO2521-ORMDL3-GSDM locus on human chromosome 17q12-q21 and the ZPBP ZNFN1A1-FIGNL1-DDC-GRB10-COBL-SEC61G-EGFR-LANCL2 locus on human chromosome 7p12 p11 were next compared. Comparative genomics revealed that ZPBPL-ZNFN1A3-GRB7 ERBB2 and ZPBP-ZNFN1A1-GRB10-EGFR loci were paralogous regions within the human genome. This is the first report on identification and characterization of the ZPBPL gene. PMID- 12883659 TI - Identification and characterization of LASP2 gene in silico. AB - LASP1 (also known as MLN50) gene, located centromeric to the PPP1R1B-ERBB2-GRB7 locus on human chromosome 17q12, is amplified and over-expressed in breast cancer. Here, we identified and characterized a novel LASP1-related gene, LASP2, by using bioinformatics. Nucleotide sequence of human LASP2 cDNA was determined in silico by assembling EST BF699808 and 5'-truncated FLJ39221 cDNA. Nucleotide sequence of mouse Lasp2 cDNA was derived from 1200007O21Rik cDNA. Human LASP2 (270 aa) showed 97.4% and 63.7% total-amino-acid identity with mouse Lasp2 and human LASP1, respectively. LASP2 and LASP1 were the LASP family proteins consisting of LIM domain, Nebulin repeat, and SH3 domain. LASP2 and NEBL mRNAs were transcribed from the LASP2/NEBL gene on human chromosome 10p12 due to alternative splicing. LASP2 mRNA consists of exons 1a-4a, 24, 27, and 28 of the LASP2/NEBL gene, while NEBL mRNA consists of exons 1-28. Exon 1a-4a of the LASP2/NEBL gene were more homologous to exon 1-4 of the LASP1 gene on human chromosome 17q12, while exon 1-28 of the LASP2/NEBL gene were more homologous to exons of NEB gene on human chromosome 2q23. Some part of the LASP2/ NEBL-TEM7L ARL8-CACNB2 locus on 10p12 was paralogous to the LASP1-TEM7-CACNB1 locus on 17q12, while the other part of the LASP2/NEBL-TEM7L-ARL8-CACNB2 locus was paralogous to the NEB-ARL5-CACNB4 locus on 2q23. These facts indicate that the LASP2/NEBL-TEM7L-ARL8-CACNB2 is a chimeric locus, which might be generated through the homologous recombination between the ancestral lasp2-tem7l-cacnb2 locus and the ancestral nebl-arl8 locus. Therefore, gene fusion during evolution is one of the mechanisms to generate alternative splicing. PMID- 12883660 TI - Differential expression of neuropilin-1 in malignant and benign prostatic stromal tissue. AB - Neuropilin-1 (NRP-1), a co-receptor for VEGF165, is overexpressed in various prostate cancer cell lines and in advanced prostate tumors. However, distribution of the NRP-1 in prostate tumors has not yet been evaluated. Using immunohistochemical analysis, we evaluated 21 archival prostate tumors and 5 benign glands for the expression of NRP-1. In addition, we utilized a quantitative RT-PCR method to examine mRNA expression in 9 additional prostate tumors obtained from radical prostatectomy specimens and compared this expression to the adjacent normal tissue. The RT-PCR analyses demonstrated overexpression of NRP-1 mRNA in malignant tissue samples by 10.0-fold as compared to adjacent normal tissue. By immunohistochemistry, NRP-1 protein was undetected or minimally detected in the epithelial tumor cells. However, NRP-1 immuno-reaction was detected in the surrounding tumor stroma. Variable immuno-reaction for NRP-1 was also seen in the adjacent normal tumor stroma and the stroma of the benign prostate samples. These observations suggest that neuropilin-1 is expressed in the prostatic stromal cells, not epithelial tumor cells, and this expression is significantly increased in the malignant phenotype. PMID- 12883661 TI - Correlation of cyclooxygenase-2 pathway and VEGF expression in head and neck squamous cell carcinoma. AB - The purpose of this study was to determine whether cyclooxygenase-2 (COX-2) was overexpressed in head and neck squamous cell carcinoma (HNSCC) in relation to vascular endothelial growth factor (VEGF) expression and lymph node metastasis. COX-2 has an important role in the promotion of carcinogenesis, tumor invasiveness and angiogenesis. VEGF is a proangiogenic factor that is up regulated in various tumors. VEGF has been shown to interact with COX-derived prostaglandins in angiogenesis. To better understand the roles of these genes in HNSCC, we evaluated the immunohistochemical expression pattern of COX-2 and VEGF in relationship with histologic differentiation, clinical stage, and nodal status in 146 HNSCC. The COX-2/VEGF double immunofluorescein staining was evaluated with confocal scanning laser microscope. A positive expression was seen in 96% (140/146: 98 strong positive, 42 weakly positive) for COX-2 and in 98% (143/146: 63 strong positive, 80 weakly positive) for VEGF in HNSCC. A correlation was present in the positive expression of COX-2 with histologic differentiation, clinical stage, and nodal status. VEGF expression was also correlated with nodal status. A significant relation was observed between COX-2 and VEGF expressions. The simultaneous expression of COX-2 and VEGF was statistically significant (p<0.05). These results indicate that elevated COX-2 or VEGF expression is more common in HNSCC with poor prognostic characteristics. The present findings support the efforts to initiate clinical trials on the efficacy of COX-2 inhibitors in adjuvant treatment of HNSCC. PMID- 12883662 TI - Thymidylate synthase expression as a predictor of the prognosis of curatively resected colon carcinoma in patients registered in an adjuvant immunochemotherapy clinical trial. AB - The expression levels of thymidylate synthase (TS) affect the sensitivity of tumor cells to fluorinated pyrimidine cytotoxic agents and determine the response of patients with colorectal cancer to fluorinated-pyrimidine-based chemotherapy. The correlation between the expression of TS and the prognosis of patients with colorectal cancer was examined in a prospective study. Evaluation of biomarkers including TS expression was performed using tumor specimens from 229 colorectal cancer patients. Immunohistochemical analysis of TS expression was performed using an antibody raised against a C-terminal epitope (D186-V313) of the human TS. The intensity of TS staining and the expression of other biomarkers were blindly scored. The five-year survival rates were 63.4% and 85.6% among patients with high and low intratumoral TS expressions, respectively (p=0.0007). Similarly, the disease-free survival (DFS) rate was 51.2% and 80.3% in the high and low TS expression groups, respectively (p=0.0004). In a subset analysis of Dukes' stage C patients, the survival and DFS rates were 44.0% and 40.0% in the high TS expression group, and 73.5% and 67.4% in the low TS expression group, respectively. Significant differences were observed in both survival (p=0.0048) and DFS (p=0.0054) rates. No significant correlation was observed between the expression of other biomarkers and prognosis. Significantly poorer prognosis of curatively resected colon cancer in patients with high TS expression levels in tumor tissue was confirmed by a double-blind prospective study conducted on samples obtained from patients enrolled in an adjuvant immunochemotherapy randomized clinical trial. PMID- 12883663 TI - Prognostic value of p53 and Ki-67 expression in resected or biopsy specimens of bile duct carcinoma. AB - For cases of inoperable bile duct carcinoma, we perform intraluminal irradiation using an 192iridium wire following endoprostheses implantation. However, the effectiveness of this procedure is uncertain, and may lead to decreased patient quality of life in some cases. Therefore, we obtained samples of bile duct carcinoma either by percutaneous transhepatic cholangioscope (PTCS) or by surgery, and studied whether expression levels of Ki-67 and p53 in these tissues could predict the effectiveness of radiotherapy (RT). Immunohistochemistry was used to determine the expression of p53 and Ki-67 in 40 resected and 18 biopsy specimens. All biopsy specimens were stage IVA according to UICC classification. Labeling indices were calculated as percentage of positively stained tumor cell nuclei of total tumor cells counted. Samples were divided into two groups according to labeling index (LI). In the resected specimens, Ki-67 LI was significantly higher in cases positive for lymphatic invasion than in negative cases (p<0.05), or advanced-stage cases (p<0.05). Also, mean Ki-67 LI was higher in tumors from cases with lymph node metastasis than without. In the biopsy specimens, a significant correlation between Ki-67 LI and the term of stent patency (p<0.05) was observed. However, there were no significant correlations between clinicopathological factors or stent patency and p53 immunoreactivity. Assessment of mean Ki-67 antigen expression, as measured by MIB-1 staining, in samples of hilar bile duct carcinoma appeared to be an important indicator of clinical behavior. Biopsy specimens obtained by PTCS may be very useful in predicting the effectiveness of RT and assist in the selection of appropriate therapies. PMID- 12883664 TI - Immunostaining of stage-specific embryonic antigen-4 in intratubular germ cell neoplasia unclassified and in testicular germ-cell tumors. AB - Stage-specific embryonic antigen-4 (SSEA-4) is expressed in testicular germ-cell tumors (GCT) according to studies using thin-layer chromatography (TLC) immunostaining. To further understand the relationship between SSEA-4 and the histogenesis of testicular GCT, we examined the expression of SSEA-4 in 43 samples of testicular GCT either by immunohistochemical staining or by TLC immunostaining. Immunohistochemical staining detected SSEA-4 in spermatogonia from the non-tumor parts of the testis and in all of 8 samples of intratubular germ cell neoplasia unclassified (IGCNU). Immunohistochemical staining was SSEA-4 positive in all 9 seminomas, and in one yolk sac tumor and in 5 embryonal carcinomas among 9 non-seminomas. Immunostaining with TLC showed that SSEA-4 was retained in 15 of 16 seminomas and in 4 of 8 non-seminomas. These results demonstrate an antigenic link between spermatogonia, IGCNU, and testicular GCT. We suggest that SSEA-4 is associated with the histogenesis of testicular GCT. PMID- 12883665 TI - Thymidine phosphorylase expression correlates with malignant potential and anti tumor effect of doxifluridine on gastric cancer: multivariate analysis for adjuvant chemotherapy doxifluridine vs. 5-fluorouracil. AB - Doxifluridine (5'-DFUR) is an anticancer drug converted to 5-fluorouracil (5-FU) by thymidine phosphorylase (TP). TP is an angiogenetic and platelet-derived endothelial cell growth factor. We evaluated the relation between TP expression and chemotherapeutic efficacy and prognosis for gastric cancer. Advanced gastric cancer patients given oral adjuvant chemotherapeutics either 5'-DFUR; 163 patients or 5-FU; 162 patients were examined. TP expression was assessed with immunohistochemical staining. Multivariate analysis for influencing survival was done, employing variables such as gender, age, procedure, tumor size, location, Borrmann type, histologic factors [type, depth of invasion, lymph node metastasis (n), lymphatic invasion (ly), and venous invasion (v)], drug administered, and TP expression. In the patients with serosal invasion, 5'-DFUR in TP positive was an independent prognostic factor (risk ratio, 4.450; 95% confidence limit, 2.099 9.436), indicating significantly improved prognosis over the 5-FU group. In TP negative, n and ly were independent prognostic factors, but the survival curves of the two chemotherapeutic groups were not significantly different. TP expression was not prognostic factor in 5'-DFUR group, while, in 5-FU group, TP expression was an independent prognostic factor (2.834, 1.467-5.476). In conclusion, it was suggested that TP positive gastric cancer with serosal invasion increased malignant potential of the tumor and 5'-DFUR efficacy. PMID- 12883666 TI - Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL/APO2L) and its receptors expression in human squamous cell carcinoma of the oral cavity. AB - Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL/APO2L) is a novel member of the tumor necrosis factor cytokine family and a potent inducer of apoptosis in tumor cells. TRAIL is expressed in most normal human cells and tissues, including peripheral blood leukocytes, spleen, lung and prostate. However, TRAIL expression in human neoplasms is largely unknown. In this study, we investigated whether TRAIL and its receptors are expressed in human oral squamous cell carcinomas (HOSCCs) or HOSCC cell lines (HSC-2, HSC-3, HSC-4, Ca9 22 and KB) and whether these cells are sensitive to TRAIL-induced apoptosis. TRAIL and its receptor transcripts and proteins were detected in all HOSCC cell lines. A cell viability (MTT) assay performed after exposure to recombinant human (rh) TRAIL for 16 h showed that KB cells were the most sensitive. Immunohistochemical examinations for TRAIL expression were also carried out in a total of 50 HOSCC cases with various types of differentiation. In 17 of 27 WHO Grade 1 SCCs, the tumor invasive areas with keratinization showed diffuse and moderate TRAIL expression. In 6 of 20 WHO Grade 2 SCCs, TRAIL expression was slightly to moderately detected. In 1 of 3 WHO Grade 3 SCCs, a slight TRAIL expression was observed. Thus, there is considerable heterogeneity of TRAIL expression and susceptibility to TRAIL-induced apoptosis among human oral tumors. PMID- 12883668 TI - Analysis of angiogenic profiles by estimation of tumor blood flow with colored dye extraction microspheres after antiangiogenic therapy. AB - No study has directly estimated tumor blood flow after antiangiogenic therapy. Using dye extraction colored microspheres, we estimated blood flow in a rat LY80 tumor model before and after antiangiogenic therapy with TNP-470. Dyed microspheres were infused into the tumor-bearing rats. The dye was extracted and its concentration was quantified by spectrophotometry. Tumor blood flow corrected for wet weight (ml. min(-1)g(-1)) was calculated as follows (AU = absorbency units): (blood flow to tumor) = (AU per gram of tumor) x (reference withdrawal rate)/(AU per gram of reference blood). Tumor tissues with or without TNP-470 were also examined histologically. Tumor blood flow 1 week after transplantation could be predicted by the simple regression equation. In the groups treated with TNP-470, this simple regression equation shifted to the left, despite virtually no difference in the results of pathological examination. In LY80 tumors 2 weeks after transplantation, blood flow in tumors treated with TNP-470 was higher than without TNP-470. This technique can be used to obtain information about the angiogenic status within tumor before and after antiangiogenic therapy. These angiogenic profiles may provide important clues to optimal antiangiogenic therapy, combinations of antiangiogenic treatment and cytotoxic therapy, and schedules for combination therapy. PMID- 12883667 TI - Adenoviral p16/CDKN2 gene transfer to malignant glioma: role of p16 in growth, invasion, and senescence. AB - In gliomas, a high frequency of homozygous p16 gene deletions have been demonstrated, which are believed to be linked with malignant progression. The aim of this study was to assess the role of p16 in growth, invasion, and senescence. The human glioma cell lines U87 MG and U373 MG were transduced with Ad-p16, and cell viability was assessed by trypan blue staining. To examine the mechanism of cell growth inhibition, cell cycle analyses and annexin assays were performed. The invasive potential of Ad-p16 transduced cells was evaluated using a Matrigel invasion assay, and trimolecular complex (MMP-2/MT1-MMP/TIMP-2) synthesis was proven by zymography and Western blotting. To establish the link between p16 and cell senescence, we stained for Senescence-Associated beta-galactosidase activity. A cell proliferation assay demonstrated that Ad-p16 treatment significantly inhibits cell growth. Moreover, this cell growth inhibition was induced by cell cycle arrest, not by apoptosis. In vitro treatment of malignant glioma cells with Ad-p16 significantly decreased their invasive potential by Matrigel invasion assay. However, we were unable to demonstrate any differences in the constitutive productions and secretions of MMP-2, MT1-MMP, and TIMP-2, among the mock-treated, Ad-lacZ-transduced, and Ad-p16-transduced cells. p16 expression caused an enlargement of all cells, and these were morphologically similar to senescent cells. Staining for Senescence-Associated beta-galactosidase activity showed that the enlarged cells stained positively. Taken together these data strongly suggest that the anti-cancer effect of p16 is modulated by p16 mediated cell cycle arrest and by the induction of senescence. PMID- 12883669 TI - The expression of tissue factor correlates with proliferative ability in meningioma. AB - Tissue factor (TF) is a cell-surface glycoprotein responsible for initiating the extrinsic pathway of coagulation; this is inhibited by tissue factor pathway inhibitor (TFPI). As TF reportedly regulates tumor growth and angiogenesis, we investigated the role of TF in meningioma. Using immunohistochemical methods, we studied the expression of TF, TFPI, MIB-1 labeling index in 44 meningiomas to determine whether those factors reflect histological grade and proliferative ability. CD31 and CD68 immunostaining was used to assess vascular density and macrophage infiltration, respectively. Additionally we assessed the influence of TF on meningioma cell proliferation by MTT assay. TF was expressed in 1 of 34 (2.9%) benign, 1 of 5 (20%) atypical, and all of 5 anaplastic meningiomas. TFPI was detected in 2 benign (5.9%), 1 atypical (20%), and 3 (60%) anaplastic meningiomas. Both TF and TFPI expression was significantly correlated with the MIB-1 labeling index (LI). However, neither TF nor TFPI showed a correlation with vascular density. The density of tumor-associated macrophages was not correlated with TF or TFPI immunoreactivity. MTT assay revealed that TF increased the proliferation of meningioma cells. Although some macrophages expressed TF, a great number of the TF immunopositive parenchymal cells in the meningiomas were tumor cells. The present study suggest that the TF system reflects the proliferative ability and malignancy of meningiomas. PMID- 12883670 TI - Tissue-specific regulation of Apaf-1 expression by p53. AB - The p53 tumor suppressor is a transcription factor that regulates cell cycle and apoptosis in response to a variety of environmental stress conditions. Recent studies indicated that the apoptotic protease activating factor-1 (Apaf-1) is a direct transcriptional target of p53 in DNA damage-induced apoptosis. However, the dependence of Apaf-1 expression on p53 has not been shown in normal physiological conditions. To further investigate their relationship, we first examined Apaf-1 protein levels in seven organs including brain, heart, kidney, liver, lung, spleen, and testis of wild-type (p53+/+) and p53-deficent (p53-/-) mice by Western blot analysis. The heart and spleen of the p53-/- mice showed a reduced Apaf-1 protein expression compared with the p53+/+ controls, while other organs did not show significant difference. The results from Western blot analysis were further confirmed by immunohistochemistry. Semi-quantitative RT-PCR was then employed to determine the expression of Apaf-1 transcripts. Similarly, we detected a lower Apaf-1 mRNA level in the heart and spleen of p53-/- mice compared with the p53+/+ controls, supporting a transcriptional upregulation of Apaf-1 by p53. Taken together, these results demonstrate that p53 regulation of Apaf-1 expression is tissue-specific. PMID- 12883671 TI - Fas stimulation activates NF-kappaB in SK-Hep1 hepatocellular carcinoma cells. AB - The TNF-receptor family has a dual signaling pathway, including induction of apoptosis and NF-kappaB activation associated with cell survival. Hepatocellular carcinoma (HCC) cells express TNF-receptor family members and the signaling from these receptors induces NF-kappaB activation. However, the role of Fas in induction of NF-kappaB activation in HCC cells is not well understood. In this study, SK-Hep1, HepG2 or HLE cells were stimulated by anti-Fas agonistic antibody. Fas stimulation induced NF-kappaB activation in a dose-dependent manner in SK-Hep1 and HepG2 cell lines, but not in HLE cells. Anti-Fas agonistic antibody or the metabolic inhibitor, cyclo-heximide (CHX), failed to kill SK-Hep1 cells, but co-incubation with anti-Fas agonistic antibody and CHX was effective for induction of apoptosis. SK-Hep1 cell lines receiving Fas stimulation had increased viability, but the extent of cell proliferation was not dose-dependent. The observation suggests that Fas stimulation may contribute to HCC cell survival or proliferation. PMID- 12883672 TI - Immunohistochemical expression of hepatocyte growth factor and c-Met/HGF receptor in benign and malignant human prostate tissue. AB - Hepatocyte growth factor (HGF) has been proposed to be an autocrine/paracrine growth factor for carcinomas of various organs. We recently demonstrated that HGF produced by prostate-derived stromal cells was a paracrine growth factor that stimulated the growth of androgen-independent prostate cancer cells in vitro and in vivo. To assess possible involvement of HGF in prostate cancer, we examined the immunohistochemical expression and localization of HGF and c-Met/HGF receptor in benign and malignant human prostate tissues. In benign glands, columnar cells generally were negative for c-Met, but basal cells were stained uniformly at a high level. In high-grade prostatic intraepithelial neoplasia (PIN) and carcinoma, more than 50% of these foci were stained uniformly. There was no difference in the frequency of positively stained cells by Gleason score. The prostate stroma stained diffusely for HGF, and the staining intensity varied depending upon the amounts of smooth-muscle cells that were stained more intensely than the connective-tissue matrix. A great majority of benign columnar cells were negative for HGF whereas high-grade PIN and carcinoma foci stained focally for HGF. Hormonal ablation therapy prior to prostatectomy did not seem to alter the expression of HGF/c-Met in carcinoma cells. These results indicate that, as the degree of neoplasia progresses, epithelial cells begin to express c Met protein, that PIN and carcinoma may have developed a c-Met-HGF paracrine loop with the stroma, and that in some carcinoma foci an autocrine loop may operate with HGF expressed by carcinoma cells themselves. PMID- 12883673 TI - Cellular heterogeneity in long-term surviving cells isolated from eutopic endometrial, ovarian endometrioma and adenomyosis tissues. AB - Human endometrial tissues regenerate easily after menstruation and childbirth, suggesting the existence of endometrial stem-like cells that can survive and proliferate from a single cell over a long time. To clarify this hypothesis, limiting dilution cultures performed with eutopic endometrial, ovarian endometrioma and adenomyosis cells obtained from a patient, achieved cloning efficiencies of 13.0, 5.0, and 0.8%, respectively. These monoclonal cells survived for more than 24 months. More than 4 types of monoclonal cells were established from eutopic endometrial cells and microscopically were distinctly different from each other. Intraperitoneal injections of dispersed human eutopic endometrial cells did not cause any endometriosis-like lesions in scid mice, but those of endometrial tissue fragments did. IL-6, TNF-alpha, IL-1beta, M-CSF and HGF failed to enhance transplantation of dispersed endometrial cells to the mice. These results indicate that several types of eutopic endometrial cells survive long-term, and that simple regurgitation of eutopic endometrial stem-like cells may not induce peritoneal endometriosis. PMID- 12883674 TI - Lymph node metastasis could be predicted by evaluation of macrophage infiltration and hsp70 expression in superficial carcinoma of the esophagus. AB - Macrophages have been reported to play an important role in suppressing tumor growth, and heat shock protein 70 (HSP70) in presenting tumor specific antigens. The objective of the present study was to investigate the relationship between lymph node metastasis and these two molecular biological markers in superficial esophageal carcinoma. Subjects were 37 patients with untreated submucosal esophageal carcinoma who underwent curative surgery. Among these patients, expression of CD68-positive macrophages in the tumor stroma and expression of HSP70 in tumor cells were analyzed. The results demonstrated that expression of CD68-positive macrophages was significantly greater among patients without lymph node metastasis (p<0.0001), and a significant correlation existed between HSP70 expression and CD68 expression (p=0.0125). In superficial esophageal carcinoma, lymph node metastasis is correlated with expression of CD68-positive macrophages in tumor stroma and expression of HSP70 in tumor cells. These two markers may therefore prove useful for therapy planning. PMID- 12883675 TI - Correlation between occult neoplastic cells in the lymph node sinuses and recurrence in patients with Dukes' C colorectal cancer. AB - The aim of this study was to investigate whether it was possible to select patients with Dukes' C primary colorectal cancer who had a high risk of metastasis and recurrence. Among 434 patients who underwent surgical resection of primary colorectal cancer, 115 patients (26.5%) had Dukes' C cancer. Among them, 35 patients (30.45%) suffered from metastasis/recurrence postoperatively. Thirty two patients with recurrence for whom complete follow-up was possible (recurrence group) and 32 patients who survived for at least 5 years without recurrence (non recurrence group) were compared by immunohistochemical staining of resected lymph nodes for cytokeratin using two antibodies (AE1/AE3 and CAM 5.2). AE1/AE3 staining was positive in 93.85% of the recurrence group versus 68.85% of the non recurrence group (p=0.0250), while CAM 5.2 staining was positive in 84.45% vs. 53.15%, respectively (p=0.0152). The occult neoplastic cell count (mean +/- SD) in the peripheral lymph node sinuses was 6.28+/-5.17 vs. 2.38+/-3.03 (p=0.0002) in the AE1/AE3-positive patients from the recurrence and non-recurrence groups, while it was 5.13+/-4.84 vs. 1.53+/-2.37 (p=0.0003) in the CAM 5.2-positive patients in each group. Accordingly, the immunohistochemical positivity rate for both AE1/AE3 and CAM 5.2, as well as the occult neoplastic cell count, were significantly higher in the recurrence group. These results suggest that patients with Dukes' C primary colorectal cancer who have a higher risk of recurrence can be selected by immunostaining of resected lymph nodes for cytokeratin. PMID- 12883676 TI - A novel phenoxazine derivative suppresses proliferation of human endometrial adenocarcinoma cell lines, inducing G2M accumulation and apoptosis. AB - We examined the effects of a novel phenoxazine, 2-amino-4,4alpha-dihydro-4alpha,7 dimethyl-3H-phenoxazine-3-one (Phx), which was produced by the reaction of 2 amino-5-methyl-phenol with bovine hemoglobin on the proliferation of human endometrial adenocarcinoma cell lines, EN and KLE cells, and on induction of apoptosis and G2M arrest in these cells. Phx inhibited proliferation of these cell lines in a dose- and time-dependent manner, i.e., the inhibition rate of proliferation of EN and KLE cells was 43% and 40%, respectively, in the presence of 50 micro M Phx, and 75% and nearly 100%, in the presence of 100 micro M Phx, after 2 days. When these endometrial adenocarcinoma cells were incubated with a medium containing 100 micro M Phx for 24 h, accumulation of EN and KLE cells in the S and G2M phase and that of apoptotic cells were demonstrated by flow cytometry. Apoptosis of these cells caused by Phx was unlikely to be associated with p53, Bax, and Bcl-2, because the levels of these proteins were not altered regardless of the presence or absence of Phx. The present results suggest that Phx demonstrates antitumor activity against human endometrial adenocarcinoma cell lines EN and KLE cells, by inducing both cell cycle accumulation at S and G2M and apoptosis associated with p53, Bcl-2 and Bax insensitive pathways. PMID- 12883677 TI - Correlation between occult neoplastic cells in the lymph node sinuses and recurrence in patients with curatively resected Dukes' B colorectal cancer. AB - This study investigated whether it is possible to detect patients who have a high risk of metastasis and recurrence after resection of stage II Dukes' B primary colorectal cancer. Among 434 patients who underwent curative resection of primary colorectal cancer, 167 (38.5%) had Dukes' B cancer. Among them, 19 patients (11.4%) suffered from postoperative metastasis or recurrence. In 17 patients with recurrence who could be followed-up completely (recurrence group) and 17 other patients who survived for at least 5 years without recurrence (non-recurrence group), immunohistochemical staining of resected lymph nodes for cytokeratin (AE1/AE3 and CAM 5.2) was performed. AE1/AE3 was positive in 76.5% and 47.1% of the patients from the recurrence and non-recurrence groups, respectively, while CAM 5.2 was positive in 52.9% and 17.6%, respectively. There were no significant differences of either AE1/AE3 or CAM 5.2 positivity between the groups. However, the occult neoplastic cell count (mean +/- SD) floating in the lymph node sinuses was significantly higher in patients from the recurrence group who were positive for AE1/AE3 or CAM 5.2 than in patients from the non-recurrence group (6.12+/ 6.00 vs. 0.59+/-0.71; p=0.0019 and 3.94+/-5.06 vs. 0.29+/-0.69; p=0.0098, respectively). These results suggest that patients with Dukes' B primary colorectal cancer who have a higher risk of recurrence can be selected by immunostaining of resected lymph nodes for cytokeratin. PMID- 12883678 TI - High risk of bile duct carcinogenesis after primary resection of a congenital biliary dilatation. AB - Congenital biliary dilatation (CBD) is a relatively rare disease and highly associated with hepatobiliary malignancies due to pancreaticobiliary maljunction. For the treatment of CBD, the standard surgical method is the excision of the entire extrahepatic duct with a hepaticoenterostomy. However, in recent years, there has been an increase in reports of cancer developing in the biliary or pancreatic duct after hepaticoenterostomy. In this report, we describe the postoperative complications and carcinogenesis in 50 CBD patients according to the method of reconstruction of the bile duct. Hepaticojejunostomy (HJ) was performed in 34 patients and hepaticoduodenostomy (HD) in 16. In the HJ group, there were 7 cases of ascending cholangitis (20.6%), 3 cases of choledocholithiasis (8.8%), 4 cases of anastomotic stricture (11.8%) and 1 case of cholangiocarcinoma (2.9%). In the patient with cholangiocarcinoma, all complications including ascending cholangitis, choledocholithiasis, and anastomotic stricture were present. In the HD group, however, 2 cases of ascending cholangitis (5.9%) were recognized although choledocholithiasis, anastomotic stricture or cholangiocarcinoma were not observed. No significant differences were found in incidences of these complications between HJ and HD. It is unclear which method of reconstruction has a higher risk of carcinogenesis, however, it can be stated that hepaticoenterostomy itself is indeed one of the risk factors for biliary tract carcinoma. In any event, as several researchers have pointed out, since the incidence of bile duct carcinoma after the resection of the extrahepatic bile duct in CBD patients is very high compared to natural control, a long-term follow-up of the patient is necessary. PMID- 12883679 TI - The dog mast cell tumour as a model to study the relationship between angiogenesis, mast cell density and tumour malignancy. AB - Substantial experimental data suggest that tumour progression is associated with angiogenesis and that increase in microvessel density (MVD) is associated with increase in mast cells density (MCD). Dog mast cell tumour (MCT) is common in dog with an incidence much higher than that found in human and in both species several common biological and clinical characteristics have been demonstrated. To evaluate the role of angiogenesis in progression of this tumour and to correlate MVD and MCD, in this study a series of 78 MCT was investigated. Serial sections obtained from biopsy specimens were processed with toluidine blue staining, specific for MC identification, and by immunohistochemistry using a polyclonal antibody anti factor VIII-related antigen (FVIII-RA), used as an endothelial marker, and MVD and MCD were determined. Results showed that MVD was significantly higher in poorly differentiated (G3) MCTs than in intermediate (G2) and well differentiated (G1) MCTs and that MCD and MVD were significantly correlated in G3, but not in G1 and G2 subgroups. These data indicate that angiogenesis and MCD are significantly correlated in MCTs progression. PMID- 12883680 TI - Mutation analysis of CTNNB1 (beta-catenin) and AXIN1, the components of Wnt pathway, in cervical carcinomas. AB - The components of the Wnt-signaling pathway are mutated in tumors, but the relationship between these components and cervical cancer has not been elucidated. In this study, we used immunohistochemistry, single strand confirmation polymorphism (SSCP) and direct sequencing methods to analyze the mutation and protein expressions of both CTNNB1 and AXIN1 in cervical cancer. Among the 30 tested cervical cancers, no mutation of CTNNB1 but 3 polymorphisms were found. Mutation analysis of AXIN1 revealed that one specimen had a heterozygous mutation at codon 740 (GCC right curved arrow ACC) and six polymorphisms were also found. Immunohistochemistry showed no relationship between the protein expression patterns and mutation of AXIN1 and CTNNB1. Mutations of CTNNB1 may not be a factor, whereas mutations of AXIN1 may play a limited role in tumorigenesis of cervical cancer. In addition, aberrant expression patterns are not mutation related, so that other factors may be responsible for these changes. PMID- 12883681 TI - Survival benefit of KRN7000 immune therapy in combination with TNP470 in hamster liver metastasis model of pancreatic cancer. AB - Pancreatic cancer is a solid malignancy with the poor prognosis largely due to frequent and lethal liver metastases. The combination of immunotherapy and anti angiogenesis therapy might be a hopeful strategy for the treatment of distant metastases. The benefits of the combination therapy by an immune stimulator alpha galactosylceramide (KRN7000) and an angiogenesis inhibitor AGM-1470 (TNP470) were evaluated on the hamster highly aggressive liver metastasis model using the syngeneic pancreatic cancer cell line HPD-NR. KRN7000 immediately activated hepatic mono-nuclear cells to produce IFN-gamma in vitro. Intraportal injection of KRN7000 exhibited a dense accumulation of CD4-CD8- natural killer T cells, around the liver metastases in vivo. KRN7000 treatment significantly inhibited the growth of liver metastases, and importantly, significant survival prolongation was confirmed when TNP470 treatment was added to it. Furthermore, cytotoxic T lymphocytes were induced at the sites of a few residual metastases in the liver of a long-term survivor. Thus, the combination of KRN7000 and TNP470 showed a high effectiveness for the treatment of liver metastases of pancreatic cancer. PMID- 12883682 TI - Occurrence of hematogenous metastasis and serum levels of thymidine phosphorylase in colorectal cancer. AB - Thymidine phosphorylase (dThdPase) is known to promote the development of new blood vessels. Increased dThdPase expression in solid tumors has been shown to correlate with tumor growth, invasion and metastasis. In the present study, we measured dThdPase levels in the tumor tissue and in the serum from the tumor drainage and peripheral venous blood obtained from patients with resectable colorectal cancer. Serum dThdPase levels, measured by a modified ELISA method, were significantly higher in patients with hematogenous metastasis. In the tumor tissue specimens, no significant difference was observed between patients with or without hematogenous metastasis. These results suggest that the serum dThdPase levels are a novel marker to predict occurrence of hematogenous metastasis in patients with resectable colorectal cancer. PMID- 12883683 TI - Impact of vasculogenesis on solid tumor growth in a rat model. AB - Peripheral stem cells released from bone marrow (BM) are known to be incorporated into foci of neovascularization and to contribute to solid tumor development. In the present rat Walker256 tumor model, BM suppression induced by total body irradiation resulted in poor growth of the tumor with apparently poor vascularization, and BM transplantation restored tumor growth. Endothelial progenitor cells are considered to be crucial for vasculogenesis, but they are not yet well defined, and methodology for their purification has not been established. As a model to examine the significance of endothelial progenitor cells in tumor-specific vasculogenesis, we utilized an immortalized rat BM derived endothelial cell line named TR-BME-2. Fluorescence-labeled TR-BME-2 cells injected systemically into rats were accumulated at the tumor site 4 days later. Another rat BM-derived cell line named C2-11, which does not have an endothelial profile, did not show tumor-specific accumulation. The tumor volume in rats treated with TR-BME-2 was significantly larger than that in rats treated with C2 11. Thus, our results suggest the importance of neovascularization by bone marrow derived endothelial cells for the promotion of tumor growth. PMID- 12883684 TI - Up-regulation and overproduction of DVL-1, the human counterpart of the Drosophila dishevelled gene, in cervical squamous cell carcinoma. AB - The Dvl-1 gene on chromosome 1p36 belongs to a family of highly conserved secreted proteins which regulates embryonic induction, generation of cell polarity and specification of cell fate through activation of Wnt signaling pathways. Wnt signaling activates the gene encoding DVL-1; the latter suppresses beta-catenin by promoting its degradation through enhanced inactivation of glycogen-synthase-kinase 3 (GSK3). Here we demonstrate increased expression of DVL-1 mRNA in over two thirds of primary cervical squamous cell cancers (11 of 15 cases) when compared to corresponding non-cancerous uterine squamous cell tissues. In addition, we noted up-regulation of cyclin D1, a downstream effector of Wnt signal pathway in cervical cancer. Immunohistochemical staining demonstrated that DVL-1 protein was prominent in the cytoplasm of cancer cells whereas it was unreactive in the surrounding normal cervical squamous cells. These data indicate that amplification and increased expression of the DVL-1 gene may play some role in the development of a portion of human cervical squamous cell cancer through derangement of the Wnt signaling pathway. PMID- 12883685 TI - Angiostatin expression in ovarian cancer. AB - Angiostatin is a potent inhibitor of neovascularization, tumor growth and metastasis. We examined the expression of angiostatin and vascular endothelial growth factor (VEGF) through immunohistochemical analysis, along with microvessel density, in primary tumors obtained from 55 ovarian carcinoma patients. Angiostatin expression was not related to either stage of disease or histology. However, VEGF expression and microvessel density were related to stage of disease. Angiostatin expression did not correlate with VEGF expression. Microvessel density correlated with VEGF, but not angiostatin expression. Univariate analysis revealed that lack of angiostatin expression, VEGF expression, microvessel density and advanced stage of disease were significant risk factors for reduced survival. Multivariate analysis revealed that lack of angiostatin expression and advanced stage of disease were significant risk factors for reduced survival. Survival time was longer in patients with angiostatin-positive and VEGF-negative tumors than in patients with angiostatin negative and VEGF-positive tumors. The presence of angiostatin expression and absence of VEGF expression are favorable prognostic factors with regard to survival in ovarian carcinoma patients. PMID- 12883686 TI - Comparison of immunohistochemistry and real-time reverse transcription-polymerase chain reaction to detect expression of carcinoembryonic antigen in lung cancer. AB - Real-time reverse transcription-polymerase chain reaction (RT-PCR) can detect mRNA of carcinoembryonic antigen (CEA) quantitatively. We compared the serum concentration, localization, and mRNA expression of CEA to determine the relationship between these three factors in patients with lung cancer. Tumors from ten patients who underwent surgery were analyzed. The serum concentration of CEA was measured before initiating therapy using a quantitative latex agglutination reaction. Immunohistochemical staining of pathologic specimens of resected tumors was performed to localize CEA. Real-time RT-PCR to detect mRNA of CEA was performed for isolating RNA from a piece of fresh-frozen tumor. Positivity for CEA production was 20% for serum, 60% for immunohistochemistry, and 80% for real-time RT-PCR. Thus, the percentage of mRNA and protein positivity of CEA in lung cancer was much higher than for the serum CEA concentration. No statistically significant correlation between the serum CEA concentration and the amount of mRNA expression was found (p=0.0932). Real-time RT-PCR is useful to quantify specific mRNA expression from a small piece of tissue. PMID- 12883687 TI - Clinical implications of intratumoral dendritic cell infiltration in esophageal squamous cell carcinoma. AB - The clinical impact of intratumoral dendritic cell infiltration (IDCI) in esophageal cancer is not fully understood. We investigated the relationship between IDCI and clinical features in esophageal carcinoma. A retrospective analysis of a total of 203 patients who underwent esophagectomy for squamous cell esophageal carcinoma was performed. We evaluated IDCI immunohistochemically using S-100-protein. Under a high power objective (x400), we evaluated IDCI in 10 regions of surgical specimen including the most invasive slice and averaged the number of S-100-protein positive cells. The patients were classified into two groups according to IDCI (>20 or <20 S-100-protein positive cells per high power field: high or low IDCI, respectively). The degree of IDCI varied from 0 to 67 (average 12.3). The 203 patients were separated into 65 with high IDCI and 138 with low IDCI. The depth of invasion was significantly more superficial in patients with high IDCI (p<0.05), and the disease was at an earlier clinical stage (p<0.01) than in those with low IDCI. Five-year survival rates after curative resection were 67% and 39% in patients with high and low IDCI, respectively (p<0.01). Multivariate analysis showed that IDCI was an independent prognostic factor (RR=1.6, p=0.02), next to nodal involvement, depth of invasion and clinical stage. Being closely related to clinical features, the prognosis of patients with esophageal cancer may be estimated by monitoring IDCI. PMID- 12883688 TI - Role of COX-2, VEGF and cyclin D1 in mammary infiltrating duct carcinoma. AB - Cyclooxygenase-2 (COX-2) has an important role in the promotion of carcinogenesis, tumor invasion and angiogenesis. Vascular endothelial growth factor (VEGF) is a proangiogenic factor that is up-regulated in various tumors. VEGF has been shown to interact with COX-derived prostaglandins in angiogenesis. Cyclin D1 gene overexpression and amplification have been shown to play a role as prognostic factors in many human cancers. To better understand the roles of these genes in mammary carcinoma, the immunohistochemical expression patterns of COX-2 and VEGF were evaluated in relationship with cyclin D1 overexpression, tumor stage, clinicopathologic parameters and patient survival in 128 mammary infiltrating duct carcinomas. The expressions of COX-2/VEGF, COX-2/cyclin D1, and VEGF/cyclin D1 were evaluated using double immunofluorescein staining with a confocal scanning laser microscope. A positive expression was seen in 41% for COX 2, 47% for VEGF, and 66% for cyclin D1 in the cases with breast cancer. There was correlation in positive expression of COX-2 or VEGF with histologic grade, lymph node metastasis, and tumor size. Conversely, a significant inverse relation was observed between VEGF and patient age. There was a correlation in overexpression of cyclin D1 with lymph node metastasis, survival rate and survival length. Significant correlations were observed between COX-2 and VEGF as well as COX-2 and cyclin D1. Co-expression of only COX-2 and VEGF was detected with significance. These results indicate that elevated COX-2 or VEGF expression or cyclin D1 overexpression is more common in breast cancer patients with poor prognostic characteristics and is partly associated with an unfavorable outcome. The present findings support the efforts to initiate clinical trials on the efficacy of COX-2 inhibitors in adjuvant treatment of breast cancer. PMID- 12883689 TI - Numerical sex chromosome aberrations in juvenile angiofibromas: genetic evidence for an androgen-dependent tumor? AB - Juvenile angiofibromas (JAs) are rare benign tumors arising almost exclusively in the posterior nasal cavity of adolescent males. While male sex predominance and tumor manifestation during adolescence are well known clinical features, their genetic causes are still unknown. Observation of an increased androgen binding rate to the cytosol of JAs and immunohistological finding of strong AR expression have suggested involvement of the androgen receptor (AR) in JA biology. In the present study, we investigated sex chromosome distribution and the expression of the AR gene in JAs of 7 males using two-color in situ hybridizations. Probes specific for the centromeres of chromosomes 1, X, and Y as well as a probe specific for the AR gene were used for investigation of paraffin-embedded JA tissue. Significant aberrations of the chromosome 1 were not observed. In 6 out of 7 analysed JAs derived from male patients we observed a significant loss of the chromosome Y in 11.5 to 63.8% (mean value: 31.3%). A gain of chromosome X was seen in 5 out of 7 JAs with the finding of two chromosomes X in 12 to 34% (mean value: 25%) of the analyzed nuclei. As each chromosome X revealed nearly almost one AR gene signal without evidence for amplifications, in 11.5 to 30% of the JA nuclei (mean value: 23.8%) two copies of the AR gene were observed. Our data indicate a significant loss of chromosome Y in combination with a gain of chromosome X in JAs. A gain of chromosome X leads to AR gene gain indicating that JAs are androgen-dependent tumors. This is supported by the finding that beta catenin known to be overexpressed in JAs acts as a co-activator of the AR. PMID- 12883690 TI - Nm23-H1 expression does not predict clinical survival in colorectal cancer patients. AB - The gene Nm23, which encodes for a nucleoside diphosphate kinase, has been defined as a metastasis-suppressor gene because of the inverse correlation between its expression and the metastatic capacity of the tumor cells. For colorectal cancer, however, the findings are equivocal. The aim of our study was to assess, in 160 patients undergoing surgery for colorectal cancer (CRC), the expression of the Nm23-H1 protein and to evaluate its possible associations with traditional clinicopathologic variables, with DNA-ploidy and proliferative activity (S-phase fraction, SPF), and with disease-free and overall survival of patients. Nm23-H1 expressions were evaluated on paraffin-embedded tissue by immunohistochemistry; DNA-ploidy and SPF on frozen tissue by flow-cytometric analysis. The median follow-up time in our study group was 71 months (range 34 115 months). No association was observed between Nm23-H1 protein expression and clinicopathological variables, S-phase fraction and DNA-ploidy. Furthermore, no significant differences were observed in the survival of patients with either moderate or strong Nm23-H1 expression. The major significant predictors for both disease relapse and death were advanced Dukes' stage, DNA aneuploid tumors and high SPF, while lymphohematic invasion was the only independent factor for relapse and non-curative resection for death. Our results indicate that Nm23-H1 activity is tissue-specific and that in CRCs the expression of the protein is not associated with tumor progression and patient prognosis, although further studies are required in order to throw more light on the possible clinical significance of the overexpression of the protein Nm23-H1 in such tumors. PMID- 12883691 TI - P53-induced gene 11 (PIG11) involved in arsenic trioxide-induced apoptosis in human gastric cancer MGC-803 cells. AB - Arsenic trioxide (As2O3) has been used as an effective chemotherapy agent for some human cancer, such as acute promyelocytic leukemia. We have demonstrated that low level of As2O3 relatively selectively inhibited growth of the solid tumor MGC-803 cells by triggering apoptosis. In this study, we found PIG11, a p53 induced gene, was upregulated markedly by As2O3 using the technique of differential display reverse transcriptase PCR (DDRT-PCR). Addition of anti-PIG11 phosphorothioated oligonucleotide (5'-GGC CGC CAT CTT CTC CTC-3') before As2O3 treatment, abolished the transient increase in PIG11 gene expression. Furthermore, it significantly inhibited the As2O3-induced apoptosis of MGC-803 cells, but had no effect in addition of missense (5'-GAG GAG AAG ATG GCG GCC-3') phosphorothioated oligonucleotides. These results suggest that PIG11, as a downstream target of p53, is involved in apoptosis of MGC-803 cells. PMID- 12883692 TI - Blockade of her2/neu decreases VEGF expression but does not alter HIF-1 distribution in experimental Wilms tumor. AB - Her2/neu regulates angiogenesis in human breast cancer, in part by stabilizing hypoxia-inducible factor 1alpha (HIF-1alpha), causing accumulation of the HIF-1 heterodimer and thus increasing expression of the proangiogenic cytokine VEGF. Her2/neu has recently been shown to be overexpressed in a subset of Wilms tumors. Using her2/neu (+) and her2/neu (-) Wilms tumor cell lines, we tested the effect of blocking anti-her2/neu antibody in vitro and in vivo. Blocking antibody did not alter VEGF expression in vitro, but decreased expression of VEGF in her2/neu (+) Wilms tumor xenografts. Tumor suppression was less marked than in parallel experiments using agents directly blocking VEGF. HIF-1alpha immunostaining was not altered in her2/neu (+) xenografts exposed to blocking antibody. These results suggest that her2/neu contributes to Wilms tumor angiogenesis in vivo by regulating VEGF, but other processes may act to rescue HIF-1alpha and thus to support tumor growth in this system. PMID- 12883693 TI - The putative serine/threonine kinase gene STK15 on chromosome 20q13.2 is amplified in human gliomas. AB - Searching for amplifications in low grade and high grade gliomas we observed an interesting correlation between the recurrence and progression of astrocytic low grade gliomas and the amplification of the STK15 gene located in the chromosomal region 20q13. Chromosome copy gains in this region have been reported previously in astrocytic gliomas and glioma cell lines and in many cancer types including breast, colorectal and ovarian cancers. The putative serine/threonine kinase STK15 has been reported to be amplified and overexpressed in breast cancer cell lines and colorectal cancer. Another candidate gene located in this region is PTPN1, a protein tyrosine phosphatase non-receptor type 1 that might play a role in cell cycle control. We used comparative PCR for quantitative DNA analysis to search for STK15 and PTPN1 amplification in gliomas previously characterized by CGH. Five out of 16 tumors (31%) of different WHO grade (1x grade II, 1x grade III, 3x grade IV) showed DNA amplification of STK15 whereas we did not detect amplification of PTPN1. We hypothesize that amplification of the STK15 gene may be a non-random genetic alteration in human gliomas playing a role in the genetic pathways of tumorigenesis. PMID- 12883694 TI - Phenotyping of IL4-induced nuclear Stat6 activity in humans: quantitation after gel shift assay using immortalized cell lines. AB - The IL4-induced Stat6 signaling pathway is active in a variety of cell types, including different cancer cells, and plays an important role in the regulation of gene expression, such as CD23. There are large quantitative differences in DNA binding activity of IL4-induced Stat6, which are useful for phenotyping activated Stat6 in normal and disease status. However, quantitation of activated Stat6 is challenging and a standardized methodology is needed. Here we have developed a semi-quantitative methodology using gel shift assay in which IL4-induced nuclear Stat6 activities are measured in human EBV-transformed lymphoblastoid cell lines. Using a DNA probe with high affinity Stat6-binding N4 motif and a specific antibody to Stat6, autoradiographs of EMSA gels are recorded by a scan imager and OD readings of antibody super-shifted Stat6 complex bands are obtained. OD readings of all test cell lines are referenced to that of a standard cell line placed in every single experiment and an OD ratio is obtained for each test cell, which allows assignment of Stat6 activational phenotypes. Using this methodology, we have been able to define three Stat6 activational phenotypes termed as Stat6high (intense banding), Stat6low (medium intensity banding), and Stat6null (very low to no discernible banding). These Stat6 phenotypes correlate well with levels of CD23 expression, but not with those of HLA-DR. Pedigree analysis has revealed a Mendelian inheritance pattern for Stat6 phenotypes. The methodology is useful in association studies in human cancer and autoimmune diseases. The Stat6null phenotype may result from a defect in Stat6 signaling which has important implications with respect to the pathogenesis of cancer and Th1/Th2 cytokine imbalance in general. In addition, the defective Stat6null lines discovered here may serve as a natural human model for comprehensive study in the same way as a Stat6 knockout null animal model does. PMID- 12883695 TI - Comparing the efficacy and safety of fixed versus weight-based dosing of epoetin alpha in anemic cancer patients receiving platinum-based chemotherapy. AB - Fixed dosing is potentially more convenient than weight-based dosing for both patients and physicians. Therefore, this open-label, randomized (1:1), multicenter study was conducted to compare the effectiveness, safety, and quality of-life benefits of fixed vs. weight-based dosing of epoetin alpha in anemic cancer patients undergoing chemotherapy. Five hundred forty-six anemic patients undergoing platinum-based chemotherapy for solid malignancies were enrolled. Patients received epoetin alpha, either a fixed dose of 10,000 IU or a weight based dose of 150 IU/kg, administered subcutaneously 3 times weekly for up to 12 weeks. Endpoints were transfusion requirements over days 29-84, change in hemoglobin (Hb) level from baseline, and change in quality-of-life (QOL) scores from baseline as measured using the Cancer Linear Analog Scale (CLAS). Five hundred and thirty-two patients received at least 1 dose of epoetin alpha, and 510 of these (255 in each treatment group) were considered evaluable for efficacy. At day 84, rates for freedom from transfusion were similar between the fixed-dose and the weight-based dose group (84% vs. 87%, respectively, p=0.32), as calculated by the lifetable method. These rates were also similar between patients in the 45-63 kg weight group receiving the fixed 10,000 IU dose or 7,000 9,000 IU on a per-weight basis (83% vs. 87%, respectively), and those in the 70 100 kg weight group receiving the fixed 10,000 IU dose or 11,000-15,000 IU on a per-weight basis (85% vs. 83%, respectively). Mean Hb increases from baseline to last observation were 2.10 g/dl [95% confidence intervals (CI95) 1.85-2.35] in the 10,000 IU group (from 9.64-11.74 g/dl) and 2.06 g/dl (CI95 1.82-2.30) in the 150 IU/kg group (from 9.70-11.76 g/dl). QOL results were similar for both groups and cumulative data have been reported. For 275 patients (in both groups combined) with CLAS QOL scores both at baseline and 29-98 days thereafter, the QOL index (average of scores for the 3 QOL parameters: energy level, ability to do daily activities and overall QOL) increased by 10.4 mm (CI95 7.5-13.2), from 46.2 mm at baseline to 56.6 mm at the final observation. QOL improvements were directly associated with Hb increases (p<0.001, multiple linear regression analysis) within all chemotherapy response classes. Epoetin alpha was well tolerated in both groups. Fixed (10,000 IU) and weight-based (150 IU/kg) dosing regimens of epoetin alpha demonstrated similar efficacy in maintaining freedom from transfusion, increasing Hb levels, and improving QOL in anemic cancer patients undergoing platinum-based chemotherapy. QOL improvements were directly associated with Hb increases. These findings support the use of a fixed-dose regimen of epoetin alpha, which may offer greater convenience for physicians and patients than weight-based dosing with this agent. PMID- 12883696 TI - Frequent loss of heterozygosity on chromosome 10p15, a putative telomerase repressor/senescence gene locus, in gastric cancer. AB - Previous studies suggest that a telomerase repressor/senescence gene, which acts as a tumor suppressor gene, may be located on chromosome 10p15. However, there are no studies on alterations on chromosome 10p15 in gastric cancers. We, therefore, examined loss of heterozygosity (LOH) on the 10p15 in gastric cancers by microsatellite assay. Two microsatellite loci, D10S501 and D10S602, were used. Fifty-seven gastric cancers, including 36 intestinal type and 21 diffuse type, were selected. LOH at D10S602 and D10S501 was detected in 6 of 18 (33%) and 5 of 27 (19%) gastric cancers, respectively. There was no significant correlation between LOH at these loci and clinicopathologic features, including patient age, sex, tumor location, histologic subtype, depth of invasion, and lymph node metastasis. These data suggest that a putative telomerase repressor/senescence gene may be located on chromosome 10p15, especially at the D10S602, in gastric carcinogenesis, and that the putative gene malfunction may be involved in the early stages of gastric carcinogenesis. PMID- 12883697 TI - Cell growth- and P53-dependent transcriptional activity of the midkine promoter confers suicide gene expression in tumor cells. AB - Midkine (MK) is preferentially expressed in a number of human tumors, while the expression in adult normal tissues is restricted. Previous studies showed that a 2.3-kb regulatory region of the human MK gene could selectively activate a linked suicide gene in tumors. In this study, we explored the minimal promoter region using genomic fragments deleted from the 5'-upstream side and analyzed the mechanism of the preferential activation in tumor cells. Luciferase assays showed that the 0.3-kb fragment from the transcription start site contained a cis-acting element(s) for the promoter activity. Expression of the herpes simplex virus thymidine kinase gene under the control of the MK promoter followed by ganciclovir administration produced antitumor effects in vivo. Transfection of the wild-type p53 gene into the immortalized fibroblasts bearing mutated p53 and tumor cell lines, which induced cell cycle arrest, decreased the MK promoter mediated transcription more effectively than the SV40 or the cytomegalovirus promoter-mediated transcription. The P53-mediated downregulation of the MK promoter activity was stronger in p53-defective tumors than in wild-type p53 bearing tumors. Moreover, the MK promoter-mediated luciferase activity was greater in p53-deficient mouse embryonic fibroblasts than in those bearing wild type p53 gene. The transcriptional activity of the MK promoter could be regulated by cell growth and in part P53-dependent pathways. PMID- 12883698 TI - Induction of antigen specific cellular immunity by vaccination with peptides from MN/CA IX in renal cell carcinoma. AB - We investigated the induction of the specific immunity for renal cell carcinomas (RCC) using MN/CA IX, a tumor-associated antigen frequently expressed in RCC. We have generated 9-mer peptide derived from MN/CA IX and examined the antigenicity as a vaccine to induce specific immunity for RCC. To use mouse syngeneic system, we transfected human MN/CA9 cDNA into RenCa and BALB-3T3 cells originally from BALB/c mouse, and established MN/CA IX expressing mouse cell lines, i.e., MN RenCa and MN-3T3. The immunization of BALB/c mouse with MN-RenCa cells resulted in the induction of cytotoxic T lymphocytes (CTL) against MN/CA IX expressing cells and the CTL clone was established from bulked CTL. This CTL clone specifically lyzed MN-3T3 cells, but not parental cells. To identify the targeted epitope binding to H-2Kd antigen, three 9-mer peptides (A, B, C-peptide) of human MN/CA IX compatible with the H-2Kd as well as HLA-A24 binding motif was synthesized. The cloned CTL targeted the B-peptide pulsed BALB-3T3 cells as well as MN-3T3 cells. Furthermore, spleen cells from BALB/c mouse immunized with B peptide reacted against MN-RenCa cells. These results suggest that the peptides derived from MN/CA IX containing HLA-A24 binding motif may be useful as a potent tumor vaccine for the treatment of human RCC, and in mouse models. PMID- 12883699 TI - CD34-positive stromal cells in primary lung carcinomas. AB - To investigate the phenotypes of stromal cells in primary lung carcinomas, we examined the distribution of CD34-positive stromal cells in primary lung carcinomas, with special reference to histological types. In total 26 surgically resected primary lung carcinomas (13 adenocarcinomas, 10 squamous cell carcinomas, 2 large cell carcinomas, and 1 small cell carcinoma and their normal tissues were examined. CD34-positive stromal cells were observed in connective tissue adjacent to the bronchiolar and bronchial epithelium in normal lung tissues. Nine of the 13 adenocarcinomas had CD34-positive stromal cells in the tumor stroma, whereas none of the other histological type tumors examined had CD34-positive stromal cells. These results suggest that CD34-positive stromal cells are specific in the stroma of primary lung adenocarcinomas, and there is a possibility that CD34-positive stromal cells may play a supportive role in primary lung adenocarcinomas. PMID- 12883700 TI - High prevalence of hypothyroidism among patients with cutaneous melanoma. AB - We have previously reported a high prevalence of hypothyroidism among patients with uveal melanoma. The objectives of the present study were to determine if a similar pattern of thyroid pathology exists among patients with cutaneous melanoma as well. To address this question, the medical records of all patients registered at the University of Texas M.D. Anderson Cancer Center with a diagnosis of cutaneous melanoma during the years 1997 and 1998 were examined for a history of overt hypothyroidism, defined as a requirement for thyroid hormone replacement. Data regarding stage and site of the primary tumor were obtained for these patients and for age/gender matched euthyroid controls from the same melanoma study population. Among 1,580 cutaneous melanoma patients (948 M/632 F), 111 (7.0%) gave a history of hypothyroidism [23/948 M (2.4%) and 88/632 F (13.9%)]. The prevalences of hypothyroidism for both males and females were significantly higher than those reported for the general population. Characteristics of the primary tumor did not differ between cases and controls, although there was a trend for a lower rate of primary tumor ulceration among the hypothyroid case subjects. We conclude that hypothyroidism of varied etiologies is common among patients with cutaneous melanoma. These data suggest that melanoma may be responsive to hormones of the thyroid hormone control loop, raising many questions of clinical and biologic importance. PMID- 12883701 TI - Factors that influence the incidence of microscopic carcinoma in frozen and preserved specimens from patients with breast carcinoma after breast-conserving surgery. AB - Because a positive margin is an important risk factor for decreased survival of patients with breast cancer after breast-conserving surgery (BCS), we examined the rate of positive surgical margins after BCS, determined from intraoperative frozen and preserved sections, and we assessed factors that influence the incidence of positive margins in surgical specimens. The cases of 172 patients who underwent BCS between June 1989 and March 2001 were studied. The pathologic features and positive margins were evaluated by microscopic examination of frozen and preserved sections. The grade of each positive margin was arbitrarily classified into one of four groups: i) cancer involved; ii) closed margin (<2 mm); iii) partially closed margin (>3 mm and <5 mm) and iv) free margin (>5 mm). Biomarkers were estimated by immunohistochemistry and DNA ploidy, and the relationship of clinical parameters was statistically analyzed using the chi(2) test. In 59 of 172 cases (34.3%), the surgical margin was positive: horizontal in 47 cases (27.3%) and vertical in 12 cases (6.9%). The free margin was significantly correlated with positive status of estrogen and progesterone receptors and middle age (35-50 years) in the patients, while clinicopathologic factors were not correlated with positive margins in surgical specimens. Although 17 cases of cancer were found in 9 frozen specimens and 8 preserved sections, the mammographic findings showed spiculated masses in 7 of the 9 frozen specimens (77.7%) and 4 of the 8 preserved sections (50%), and microcalcification in 2 frozen specimens (22.2%) and 4 preserved sections (50%). The overexpression (2+ or 3+) of the HER-2 protein was found in 8 of the 9 frozen specimens (87.5%) and in 7 of the 8 preserved sections (88.8%). In the 11 cases with spiculated masses, extensive intraductal components (EICs) of various grades were found; the presence of EICs in a spiculated lesion was observed in 5 of the 7 frozen specimens and in all 4 of the preserved sections. The local extension of breast carcinoma cells in BCS is associated in part with the findings of spiculation and microcalcification, which are associated with EICs. The overexpression of HER-2 protein appears to play a role in EICs in positive margins, and needs to be considered for avoiding positive margins in BCS on patients with breast carcinoma. PMID- 12883702 TI - Relationship between AgNORs, MIB-1 and oncogene expression in male breast carcinoma and papillary superficial bladder neoplasm. AB - The expression of p53, c-erbB-2, bcl-2 and c-myc proteins was compared to the quantity of the nucleolar organiser regions (AgNORs) and MIB-1 antigen to elucidate the relationship between oncogene expression and rapidity of cell proliferation and tumor growth fraction. Sections from 50 male breast carcinomas (MBC) and 62 superficial papillary bladder neoplasias were stained with the standardised AgNOR method and monoclonal antibodies MIB-1, DO7, CB11, bcl-2 124 and 9E11. p53 immunopositivity was associated with high AgNOR quantity and MIB-1 scores both in MBC and bladder neoplasm. c-erbB-2 expression was associated with high AgNOR quantity in bladder neoplasm. bcl-2 expression was associated with low AgNOR quantity in MBC. c-myc expression was associated with high AgNOR quantity in MBC. MBC patients with low AgNOR quantity, and p53, c-erbB-2 and c-myc immunonegativity had the longest overall survival. Patients with bladder neoplasia with low AgNOR quantity, negative p53 and positive c-erbB-2 immunostaining had the longest disease-free survival time. Our results indicate that p53 overexpression reflects both the rapidity of cell proliferation, as assessed by AgNOR quantity, and tumor growth fraction, as assessed by MIB-1 scores, while c-erbB-2, c-myc and bcl-2 expression mainly reflects the rapidity of cell proliferation. The combination of AgNOR quantity and oncogene expression may stratify patients into different risk groups. PMID- 12883703 TI - Survivin expression is significantly linked to the dedifferentiation of thyroid carcinoma. AB - Survivin is a novel member of the apoptosis protein inhibitors, but according to previous reports, it is also significantly linked to cell proliferating activity. In this study, we investigated the expression of survivin in thyroid neoplasms. Survivin was only occasionally expressed in normal follicular cells, whereas in follicular and papillary carcinomas, about 20% of cases were positive for survivin. The incidence was significantly higher in advanced stage papillary carcinoma (p=0.0080) and papillary and follicular carcinomas with poorly differentiated lesions (p=0.0150). In anaplastic carcinoma, survivin positivity was observed in 84% of the cases, which was in significantly higher incidence than in papillary or follicular carcinoma (p<0.0001). These results suggest that survivin is strongly related to the dedifferentiation of thyroid carcinoma. PMID- 12883704 TI - Methylenetetrahydrofolate reductase gene C677T and A1298C polymorphisms in patients with small cell and non-small cell lung cancer. AB - Two mutations of methylenetetrahydrofolate reductase (MTHFR) gene (C677T and A1298C) may lead to a decreased activity of the enzyme. These mutations may change a risk of some cancers. We evaluated these two polymorphisms of MTHFR in patients with small cell lung cancer (SCLC) and non-small cell lung cancer (NCSCL). All lung cancer patients had statistically significantly higher percentage of MTHFR 677TT genotype in comparison with non-cancer controls. There were no statistically significant differences in the distribution of MTHFR 1298 genotypes. Neither of the polymorphisms presented any statistically significant differences between SCLC and NSCLC. PMID- 12883705 TI - Combined use of established and novel tumour markers in the diagnosis of head and neck squamous cell carcinoma. AB - Serum SCC, CYFRA 21-1, and CEA are the common tumour markers for head and neck squamous cell carcinoma (HNSCC), although diagnostic sensitivity should be yet improved, especially at early stages. In the present study, we have reported the diagnostic value of two novel serum tumour markers in HNSCC: alpha-L-fucosidase (AFU) activity, and total sialic acid concentration adjusted by total protein concentration (TSA/TP). Using the cut-off 4.0 U/ml, AFU showed a sensitivity of 55% with specificity levels of 91%, 85% and 50% to discriminate HNSCC patients from healthy donors, drinking and smoking subjects, and patients with benign diseases, respectively. Furthermore, AFU showed the best sensitivity (71%) in the detection of patients with premalign lesions. Using the cut-off 12.0 ng/mg, TSA/TP showed the best sensitivity levels (63%) in the diagnosis of HNSCC with specificity levels of 94%, 50% and 90%, regarding healthy donors, drinking and smoking subjects, and patients with benign diseases, respectively. It was of special interest that sensitivity in the diagnosis of HNSCC at non-disseminated stages was improved when using combinations of AFU+CYFRA or TSA/TP+CYFRA, up to 86% or 71% in TNM I, 60% or 80% in TNM II, and 80% or 60% in TNM III, respectively. PMID- 12883706 TI - Rho regulates the hepatocyte growth factor/scatter factor-stimulated cell motility of human oral squamous cell carcinoma cells. AB - To investigate the effects of hepatocyte growth factor/scatter factor (HGF/SF) on the invasion and metastasis of human oral squamous cell carcinoma (SCC) cells, we examined cell motility and intercellular signal transduction of a human oral SCC cell line (SAS) obtained from the primary lesion of a tongue carcinoma. HGF/SF stimulation significantly enhanced the motility of SAS cells in a dose-dependent manner. Clostridium botulinum C3 exoenzyme (C3), which is known to selectively impair the function of Ras-related small G-protein p21rho (Rho), significantly reduced the motility of SAS cells. HGF/SF stimulation also enhanced the tyrosine phosphorylation of HGF receptors (c-Met) and focal adhesion kinase (FAK) on SAS cells, but C3 completely inhibited the phosphorylation of FAK. Furthermore, it was observed that Rho A protein, normally located around the nuclear area, was translocated to the membrane and levels in the cytolysate increased following HGF/SF stimulation with no change in Rho A mRNA. These results suggest that the activation of FAK caused by phosphorylation of c-Met may mediate the HGF/SF induced motility of human oral SCC cells, and that Rho protein regulates the tyrosine phosphorylation of FAK through translocation from the nucleus to the membrane. PMID- 12883707 TI - VHL tumor suppressor gene: its mutation and protein level in renal cell carcinoma. AB - In this study, we identified mutations in the von Hippel-Lindau gene (VHL) in patients with sporadic clear cell renal carcinoma and analyzed the relationship between these VHL mutations and VHL protein (pVHL) expression levels. Analysis of VHL coding region mutations by direct polymerase chain reaction in 39 patients with clear cell renal carcinoma, who had all undergone radical nephrectomy, revealed heterozygous mutations in 7 (17.9%) of the 39 patients, including 2 novel mutations. Analysis of the levels of pVHL expression in tumor tissue samples from the 7 patients with mutations by Western blot analysis demonstrated that pVHL was not expressed in 3 patients, and that 3 subjects expressed about 50%, and 1 was similar to the normal level. These results suggest that the 3 patients who did not express pVHL had both mutation and loss of heterozygosity, and that the 3 patients who expressed about 50% of the normal pVHL level had heterozygous mutations. PMID- 12883708 TI - Ultrastructural change due to acquired cisplatin resistance in human bladder cancer cells. AB - Cis-diaminedichloroplatinum (cisplatin) has therapeutic efficacy against advanced bladder cancer. However, the response is often limited due to appearance of drug resistant tumor cells. Recently, we established a cisplatin-resistant human bladder cancer cell line, T24/R2, from T24 human bladder cancer cell line, which shows 18-fold resistance to cisplatin by the stepwise exposure of increasing concentrations of cisplatin. A light microscopic (LM) and transmission electron microscopic (EM) examination was performed to investigate the morphological and ultrastructural changes during induction of drug resistance. In LM, the cytoplasm of T24R2 cells showed plumper pattern than that of T24 parent cells. In EM, the chromatin pattern of T24R2 cells was finely dispersed compared to T24 cells, which were coarse and aggregated. The mitochondrial volume, rough endoplasmic reticulum, polyribosomes and ribosomes were moderately increased in T24R2 cells. The cell membrane showed ruffled border and great amount of double membrane vesicles and pinocytic vesicles were observed in the cell surface of T24R2 cells, which were seldom observed in T24 cells. With these findings, we concluded that human bladder cancer cells underwent morphological and ultrastructural changes during acquiring of resistance to cisplatin. We could suggest that these changes might be involved in the drug resistance mechanism in human bladder cancer cells. PMID- 12883709 TI - The copper-chelating agent, trientine, attenuates liver enzyme-altered preneoplastic lesions in rats by angiogenesis suppression. AB - It has been shown that angiogenesis plays an important role not only in tumor growth, but also in carcinogenesis. We previously reported that the copper chelating agent, trientine dihydrochloride (trientine), exerted strong anti angiogenic activity and inhibited hepatocellular carcinoma (HCC) tumor growth. The aim of the current study was to elucidate the effect of trientine on liver enzyme-altered preneoplastic lesions in rats, especially in conjunction with angiogenesis alteration in the liver. In a diethylnitrosamine (DEN)-induced rat hepatocarcinogenesis model, trientine treatment, even at a clinically comparable low dose, significantly suppressed glutathione S-transferase placental form (GST P)-positive preneoplastic lesions associated with a decrease in copper content in the liver. Trientine also markedly suppressed neovascularization in the liver to a similar level as that of development of the preneoplastic lesions. On the contrary, the proliferative cell nuclear antigen (PCNA)-positive cells were not altered with or without trientine treatment. These results suggested that the copper-chelating agent, trientine, exerted chemopreventive effects against rat liver carcinogenesis due to the suppression of angiogenesis, and suggest that it might be useful clinically as a chemopreventive agent of HCC. PMID- 12883710 TI - Reduced expression and loss of heterozygosity of the SDHD gene in colorectal and gastric cancer. AB - Mitochondrial DNA (mtDNA) mutations occur in a variety of human cancers, suggesting a possible role for mitochondrial respiratory functions in tumorigenesis. Recent studies have demonstrated that SDHD, a nuclear gene encoding one of the mitochondrial complex II subunits, acts as a tumor-suppressor for hereditary paragangliomas and pheochromocytomas. In order to determine whether the SDHD function plays a wider role in human malignancies, we examined SDHD gene alterations in 52 colorectal and 59 gastric cancers and 7 cancer cell lines. Loss of heterozygosity (LOH) at the SDHD gene locus was found in 5 of 35 (14%) colorectal and 5 of 40 (13%) gastric cancers. Reduced SDHD gene expression, which was partly associated with SDHD gene LOH, was observed in 15 of 19 (79%) colorectal cancers examined. Unlike classical tumor-suppressor genes, however, partial loss rather than complete loss of the SDHD gene expression was preferentially observed and the reduced expression could not result from CpG island methylation or coding mutation. Interestingly, the mtDNA mutations (12 cases) and the SDHD gene LOH (6 of 7 cases) did not occur in the same cancers, suggesting that these alterations might have similar functional effects in tumorigenesis. We suggest that SDHD alterations can affect mitochondrial respiratory chain functions and play a role in colorectal and gastric cancers as a distinct type of tumor suppressor. PMID- 12883711 TI - Clinical implication of cyclin B1 in non-small cell lung cancer. AB - Cyclin B1 plays an important role in the mitotic cycle, in which it regulates the G2-M transition, and has been suggested to play a role in development and progression of various cancers. The present study was undertaken in order to clarify the role of the cell cycle regulator cyclin B1 in non-small cell carcinoma (NSCLC). We retrospectively investigated 174 patients with NSCLC who previously underwent complete resection. There were two study groups: the squamous cell carcinoma (SCC) group (n=62); and the non-SCC group (n=112). Expression of cyclin B1 in cancer cells was analyzed immunohistochemically. The rate of cyclin B1 positivity in cancer cells ranged from 0% to 56.5% (average 10.9%). Seventy-four cases (42.5%) were designated as cyclin B1 positive in the present study. Cyclin B1 expression was observed more frequently in SCC cases than in non-SCC cases. In SCC, cyclin B1 expression demonstrated significant correlation with gender (p<0.01) and histological type (p<0.01). In non-SCC, only gender was correlated with cyclin B1 expression. Five-year survival rates for cyclin B1-positive and cyclin B1-negative cases were 45.8 and 57.9%, and 10-year survival rates were 39.3 and 51.4%, respectively. Patients with positive cyclin B1 staining showed a lower survival rate than those with negative staining (p=0.11). The prognostic value in SCC cases was p=0.48. In non-SCC cases, the survival rate of non-SCC patients who were positive for cyclin B1 was significantly lower than that of patients who were negative (p<0.01). Using multivariate analysis, tumor size (p=0.037) and N factor (p=0.026) were found to be independent prognostic parameters. Cyclin B1 expression was not an independent prognostic factor in the present series. These data suggest that elevated levels of cyclin B1 expression may be an indicator of poor prognosis in NSCLC, particularly in non-SCC. PMID- 12883712 TI - Expression of a 65 kDa oncofetal protein in human prostatic carcinoma. AB - The concentration of a novel 65 kDa oncofetal protein, p65, was measured in the sera of patients with prostatic malignancies using an enzyme-linked immunosorbent assay. The prostate cancer sera were positive for p65 in 40 out of 59 cases (68%), while only 7 of 79 normal sera (9%) and 12 of 61 sera from patients with benign diseases (20%) were positive. The detection system had an overall sensitivity of 68% and a specificity of 86%. Elevated p65 levels correlated positively with the pathologic stages of the prostate cancer. Using this marker in concert with PSA may increase our ability to evaluate treatment response and aid in early detection of prostate cancer. PMID- 12883713 TI - Chromosomal imbalance in esophageal squamous cell carcinoma: 3q gain correlates with tumor progression but not prognostic significance. AB - DNA copy number changes were analyzed by comparative genomic hybridization (CGH) in 53 esophageal squamous cell carcinomas (ESCC) to clarify the relationship between DNA sequence copy number aberrations and clinicopathological factors. Changes in DNA copy number were observed in all 53 ESCC patients. The average number of DNA copy number gains was 9.32 (range 1-23), most frequently located on chromosomes 3q, 5p, 8q, 11q and Xq in over 40% of tumors. Loss of DNA copy number was detected on 3p, 5q, 4p, 1p and Xp in over 20% of tumors. No statistically significant differences in the frequency of DNA copy number changes were observed. However, some loci showed correlation with clinicopathologic factors: 8q gain correlated with the pattern of tumor infiltration, 3q gain correlated with pT, 2p gain, 1p loss and 16p loss correlated with lymphatic invasion, and 3q gain correlated with clinical stage. Thus, in ESCC, gain of 3q is the only specific recurrent pattern of DNA aberration that correlates with clinicopathologic parameters, although no particular loci correlated with patient prognosis. Further CGH analysis may reveal new, recurrent genetic changes in ESCC affecting chromosomes sites that harbor genes known to participate in tumorigenesis and progression of several human malignant neoplasms. PMID- 12883714 TI - Chemotherapy-induced radiation recall myositis. AB - Myofasciitis syndrome encompasses a group of disorders characterized by chronic inflammation and/or fibrosis of the subcutaneous septa and muscular fascia. We report on a patient in whom myositis was diagnosed in the areas previously irradiated for papillary thyroid carcinoma and anal canal carcinoma respectively 21 and 3 years after radiotherapy. We are not able to explain why myopathy developed at the same time in two different sites at a different interval from the two radiotherapic schemes. We can suppose that the patient developed a subclinical regional myopathy after the first radiotherapic scheme. Radiation induced heritable mutations within surviving cells that were unable to tolerate the second damage by systemic chemotherapy. It is unclear how radiosensitization correlates with an ability to reactivate latent effects in normal tissue. Physicians using chemotherapic radiosensitizers should be aware of their potential to induce a delayed form of radiosensitization. We report this case to encourage physicians to be alert to the knowledge of the clinical, histologic and morphologic characteristics of radiation myositis in order to distinguish it from an infectious or immune fasciitis or myositis. PMID- 12883715 TI - An in vitro evaluation of radiation effects of different fractionated regimens by absolute cell count beads. AB - Recently, whole-body gamma-knife as a kind of radiotherapy equipment, characterized by precision of targeting irradiation, was applied in clinic. However, there is no conventional treatment regimen suitable for application to whole-body gamma-knife as yet. For providing reference to clinic to determine and optimize the treatment regimens for whole-body gamma-knife, a simple and quick bivariate flow cytometric approach was adopted to evaluate the killing effects of differently fractionated regimens of whole-body gamma-knife radiotherapy on the same cells, in which Annexin V-FITC (fluorescein-isothiocyanate) was used for discriminating apoptotic cells, propidium iodide (PI) for necrotic or dead cells and absolute cell count beads for absolute count of remnant surviving cells to examine the absolute killing effect. The results showed that the different survival rates of the same cells treated with different multifractionated irradiation regimens, relevant to clinical trial, might be sensitively, easily and rapidly evaluated by this approach. The evaluation of absolute killing effect, which is important to clinic, is very difficult to realize by a conventional clonogenic assay. For the approach adopted in the experiments, the cells in the treatment courses were not affected by non-treatment factors, such as cell migration, detachment, reattachment, bystander effect and medium factors encountered in a conventional clonogenic assay, and the conditions were more relevantly similar to cells in vivo. This approach might be used as a rapid cell survival assay especially in evaluating the absolute killing effect of a multifractionated treatment regimen, and also applied in the assessment in cytotoxic killing effect, such as chemotherapy. PMID- 12883716 TI - CT findings of breast cancer with clinically complete response following neoadjuvant chemotherapy--histological correlation. AB - In breast cancer patients, several regimens of neoadjuvant chemotherapy have been developed in order to achieve prognostic advantages for individual patients. Though some percentages of breast cancer patients show clinically complete response to neoadjuvant chemotherapy, the histopathological specimens of these patients demonstrate a considerably high frequency of the existence of residual disease. In this study, we aimed to evaluate the therapeutic effect of neoadjuvant chemotherapy for breast cancer patients showing clinically complete response (cCR) to neoadjuvant chemotherapy, using thin-section (5 mm) helical CT (prone position) with bolus injection of contrast agent. Between April 1994 and March 2002, 9 patients with breast cancer showing cCR to the neoadjuvant chemotherapy, who had undergone thin-section CT study both before and following neoadjuvant chemotherapy, enrolled in the study. The mean age of the patients was 46.2 years and all of them were female. The clinical stages were, 8 patients in stage II, and one in stage IIIA. In the CT evaluation, residual disease was visualized in 5 out of the 9 patients. Histopathological examination disclosed the existence of residual cancers in 6 out of the 9 patients, but only non invasive cancer was revealed in 1 out of the 6. As patients having residual disease composed only of non-invasive cancer are classified into the pathologically complete response group according to the WHO classification, 4 out of these 9 patients showing clinically complete response to the neoadjuvant chemotherapy were classified into pCR (pathologically complete response) group, and another 5 were classified into the pPR (pathologically partial response) group. As a result, the diagnostic accuracy of the second CT study performed after neoadjuvant chemotherapy was evaluated as 77.8%, with a sensitivity of 80.0%, a specificity of 75.0%, a positive predictive value (PPV) of 80.0%, and a negative predictive value (NPV) of 75.0%. Therefore, for precise evaluation of the neoadjuvant chemotherapeutic effect for breast cancer, thin-section CT studies are considered to be essential. PMID- 12883717 TI - Modulation of EGFR and neu expression by n-6 and n-9 high-fat diets in experimental mammary adenocarcinomas. AB - In this study we have investigated the influence of dietary mono- and polyunsaturated lipids on 7,12-dimethylbenz(alpha)anthracene (DMBA)-induced mammary tumorigenesis, and the modulation of expression of c-erbB1/EGFR and c erbB2/neu as a mechanism of this influence. Two series of Sprague-Dawley rats were given a single dose of DMBA. Animals from Series 1 were fed a semi-synthetic low-fat or high-fat corn oil diet, following an initiation-promotion experimental design. Rats from Series 2 were fed low-fat or different high-fat diets (rich in corn oil or rich in olive oil) in the promotion stage of mammary carcinogenesis. High corn oil diet showed clearly a stimulatory effect on experimental breast carcinogenesis, suggesting a role of this kind of lipids on initiation and promotion of mammary tumorigenesis. Moreover, results suggested that olive oil acts as a negative modulator of the experimental breast cancer. On the other hand, two transcripts of EGFR and one neu mRNA were detected by chemiluminescent Northern blot in mammary adenocarcinomas. Analysis of data showed the tendency that high-fat corn oil diet decreases the 2.7 kb mRNA of EGFR, encoding a truncated form of the receptor with no enzimatic activity. High-fat olive oil diet increased EGFR mRNA levels, specially those from 2.7 kb, and decreased the relative abundance of neu mRNA. These data suggest that the modulating effect of dietary lipids on mammary carcinogenesis could result in changes of EGFR and neu mRNA, leading to an increase of EGFR activity by high-fat corn oil diet and a decrease of EGFR and Neu signal transduction pathway by high-fat olive oil diet. PMID- 12883718 TI - Intratumoral induction of thymidylate synthase mRNA by 5-FU in colorectal cancer patients: association with survival. AB - Dihydropyrimidine dehydrogenase (DPD) is the rate-limiting enzyme in 5 fluorouracil (5-FU) catabolism. Thymidylate synthase (TS) is inhibited to form an inactive ternary complex by 5-fluoro-dUMP and is considered to be a target enzyme of 5-FU treatment. Two enzymes, DPD and TS, have been reported to be major determinants of individual sensitivity to 5-FU, and it has been reported that TS mRNA levels are modified by 5-FU treatment. We investigated their impact on treatment efficacy in colorectal cancer patients. TS and DPD mRNA levels, which correlated to the corresponding enzyme activities, were quantified in tumor tissues before and after treatment in 40 advanced colorectal cancer patients who had been treated with Doxifluridine (5'-DFUR) for 14 days before surgery. Furthermore inter-individual variations of TS mRNA levels after 5-FU treatment were found, and the individual TS induction varied between patients (0.2-2.4). Increased TS mRNA levels were found in 19 out of 40 cases. The samples were divided into two groups according to their TS mRNA induction (< or >1; TS/beta actin ratio after treatment divided by values prior treatment) and compared with tumor reduction and survival. TS and DPD mRNA levels in tumor biopsies before treatment were not related to 5-FU responses by histological evaluations in this study. However the efficacy of 5-FU treatment was enhanced in patients with no or low TS mRNA induction (odds ratio: 6.2, p<0.05). Furthermore, longer periods of survival were observed in the group without increased TS mRNA levels. These findings suggest that TS mRNA was induced by 5-FU treatment, and the overall induction level varied between individuals. Therefore, the estimation of TS mRNA induction may be useful to predict the efficacy of 5-FU treatment. PMID- 12883719 TI - E6/E7 genes of human papilloma virus type 18 induced immortalization of human fetal esophageal epithelium. AB - To study the role played by human papilloma virus (HPV) in carcinogenesis, immortalized esophageal epithelial cells were induced by E6 and E7 genes of HPV type 18 and the biological behavior was studied. Human fetal esophageal epithelial cells were transfected with recombined HPV18E6E7AAV and were cultured and passaged in medium M199. In both the 10th passage (SHEE10) and the 31st passage (SHEE31), their proliferative rates by flow cytometry and their abilities to grow and form colonies in soft agar, or to form tumors in SCID mice were examined. The HPV18 genes of E6E7 and its expression were determined using PCR methods. Cellular telomerase activity was detected by TRAP and chromosomes were analyzed by standard method. Immortalized cell lines of esophageal epithelium induced by the HPV18E6E7 were successfully established and cultured for >100 passages over 4 years. The result of PCR showed that the E6E7 gene of HPV18 was detectable in both cell clones. Both of them were unable to grow in soft-agarose medium and failed to produce tumors in SCID mice. Flow cytometry demonstrated an average of 43% proliferation index in SHEE31, but 28% in SHEE10. Telomerase activity was clearly identified in SHEE31 but not in SHEE10. Cytogenetic analysis demonstrated progression of chromosomal abnormalities with increasing trisome. Our data indicated that genes E6/E7 of the HPV18 were capable of inducing immortalization in fetal esophageal epithelial cells. The immortal phenotype requires both activation of telomerase and genetic alterations that abrogate normal differentiation and promote cellular proliferation. This cell line can assist us to characterize the role played by HPV in carcinogenesis. PMID- 12883720 TI - Topical vidarabine or 5-fluorouracil treatment against persistent HPV in genital (pre)cancerous lesions. AB - In the present study, effectiveness of topical vidarabine or subsequent 5 fluorouracil (5-FU) administration was examined against persistent genital human papillomavirus (HPV) infection after local surgery. Thirty patients underwent local eradication treatment of uterine cervical intra-epithelial neoplasia (CIN) and stage Ia1 uterine cervical cancers. HPV typing was performed by PCR-RFLP analysis. HPV infection was detected pre-operatively in 29 of 30 patients. Of these, HPV was still present in the 20 patients within two months after the therapy. Topical administration of vidarabine or subsequent 5-FU once a week for four weeks was performed to the post-operative persistent HPV-positive cases. HPV infection was abolished in 1 of 10 (10%) with topical vidarabine, and in 2 of 4 vidarabine-resistant cases (50%) with topical 5-FU. Topical vidarabine or 5-FU treatment is beneficial for HPV-positive cases after local surgical excision. PMID- 12883721 TI - The increase of CD57+ T cells in the peripheral blood and their impaired immune functions in patients with advanced gastric cancer. AB - In addition to natural killer (NK) cells, T cells expressing natural killer cell markers, CD56 or CD57 (NK type T cells), have been considered to play an important role in antitumor immunity. We examined the proportion of NK cell and NK type T cell subsets in the peripheral blood from patients with gastric cancer. The IFN-gamma production capacity and population of cytoplasmic perforin positive cells in peripheral blood mononuclear cells (PBMC) were evaluated. Peripheral blood samples were obtained from 56 patients with gastric cancer and 21 healthy volunteers. The proportion of CD56- CD57+ T cells (CD57+ T cells) was significantly higher in advanced gastric cancer patients than those in healthy volunteers and patients with early stage gastric cancer, whereas no correlation was observed between the proportion of CD56+ T cells or NK cells and tumor progression. Furthermore, a significant decrease of CD8+ CD57+ T cells was found in patients with advanced gastric cancer. The proportion of CD57+ T cells did not correlate with interferon-gamma (IFN-gamma) production from PBMC in gastric cancer patients, although a significant correlation was found between them in healthy volunteers. The proportion of perforin positive CD57+ T cells, especially CD8+ CD57+ T cells, in patients with gastric cancer was markedly lower than that in healthy volunteers. Collectively, although the proportion of CD57+ T cells in PBMC was found to increase with tumor progression, their function in antitumor immunity is impaired in patients with gastric cancer. PMID- 12883722 TI - Multidetector-row CT analysis of time-dependent changes in lung fields after chest irradiation: usefulness of precision scans. AB - Time-dependent changes in lung fields after chest irradiation were analyzed using multidetector-row CT. Routine scans at 3-mm raw thickness and 8-mm recon thickness and precision scans at 0.5-mm raw thickness and 0.5-mm recon thickness were compared with respect to the number of each finding and the time-dependent changes in the rate of each finding. Among the findings visualized by these scans, ground-glass opacity (GGO) showed the highest overall appearance rate. Precision scans exceeded routine scans in the rates of all findings except GGO and confluent shadows, and the two types of scans showed the greatest difference in the rate of GGO. Since we found that GGO tended to be overestimated on routine scans, we confirmed it by a phantom experiment. Precision scans were similar or superior to routine scans in the rates of findings except 3 months after irradiation. We consider that the concomitant use of precision scans is useful in that it allows more accurate evaluation of various post-irradiation changes in lung fields including GGO, in which the lesion is in a reversible stage. PMID- 12883723 TI - Detection of codon 61 point mutations of the K-ras gene in lung and colorectal cancers by enriched PCR. AB - Mutation of the Kirsten ras (K-ras) gene is one of most common alterations in solid tumors including lung and colorectal cancers. We developed new enriched PCR RFLP assay to detect mutations of K-ras codon 61 at the 1st and 2nd letters and non-enriched PCR-RFLP assay to detect the 3rd letter mutation. One mutant allele among 10(3) wild-type alleles was detected by enriched PCR-RFLP assay, while one mutant in 10 wild-type alleles was detected by non-enriched PCR-RFLP assay for codon 61 3rd letter. We then examined K-ras codon 12, 13 and 61 mutations in lung and colorectal cancers using these assays. K-ras codon 12 mutation was detected in 10 of 109 (9%) lung cancer and 19 of 83 (23%) colorectal cancer cases. K-ras codon 13 mutation was detected in 2 of 83 (2%) colorectal and 0 of 109 NSCLC cases, respectively. There was no K-ras codon 61 mutation in either type of cancer. Our results demonstrate that enriched PCR-RFLP is a sensitive assay to detect K-ras codon 61 mutation, however, it was extremely rare in lung and colorectal cancers, suggesting organ-specific pathways in mutagenesis of the ras gene family. PMID- 12883724 TI - Protein expression of p53 and Bcl-2 has a strong correlation with radiation resistance of laryngeal squamous cell carcinoma but does not predict the radiation failure before treatment. AB - To examine what factors can precisely predict the radio-sensitivity or radio resistance of early stage laryngeal squamous cell carcinomas (LSCC), protein expression of p53, Bcl-2 and Bax and loss of heterozygosity (LOH) at 3p and 9p loci were investigated. From June 1994 through June 1999, specimens of primary tumors were obtained by biopsy from 21 patients diagnosed as early stage LSCC at Miyagi Cancer Center Hospital under approved protocol. Labeling indexes of p53 and Bcl-2 were markedly increased in the recurrent tumors (n=8) and the differences were significant by statistical analysis (p=0.043 and p=0.015). However, when labeling indexes of the samples before treatment from non-recurrent cases were compared with those of the samples before treatment from recurrent tumors (n=13), no significant difference was observed. When LOH was examined, 16 out of these 21 cases were acceptable for evaluation, 4 of which (25%) had a relapse. Allelic loss at the 9p21 and 3p21 loci was found in 6 of 13 cases (46%) and 10 of 16 cases (63%), respectively. The frequency of LOH at the 3p21 locus in the recurrent cases was 100% (4 out of 4) and that in non-recurrent cases was 50% (6 out of 12). The frequency of LOH in recurrent tumors was apparently higher than that in non-recurrent tumors. Preservation rates of the larynx of the patients with 3p21 LOH negative tumors was very high (100%) and that of the patients with 3p21 LOH positive tumors was low. These results indicate that although expression of p53 and Bcl-2 had a strong correlation with recurrent LSCC treated by irradiation, immunohistochemical analysis of p53 and Bcl-2 in the sample before treatment was not able to predict the radiation failure. Moreover, our results also indicated that molecular research on genetic instability such as allelic loss may be more sensitive for the detection of radio-resistant tumor cells. PMID- 12883725 TI - P38 MAPK pathway is involved in the urokinase plasminogen activator expression in human gastric SNU-638 cells. AB - Overexpression of urokinase plasminogen activator (uPA) is known to correlate closely with tumor cell invasion and metastasis. In gastric cancer, however, the mechanism for induction of uPA remains to be elucidated. In this study, we investigated the intracellular signaling for uPA expression in human gastric carcinoma cells (AGS, SNU-1, SNU-5, and SNU-638). SNU-638 cells which expressed a high level of uPA was found to be highly invasive on a matrigel, while AGS, SNU 1, and SNU-5 cells with low levels of uPA expression were only slightly invasive. SNU-638 cells showed a much higher P38 MAPK activity than the 3 other cell lines. However, there was no significant difference in the activities of P44/42 MAPK (Erk-1/2), JNK and Akt among the above cell lines. Treatment of SNU-638 cells with SB203580, a specific P38 MAPK inhibitor, reduced both the promoter activity and mRNA expression of uPA. Expression of a vector encoding a mutated-type P38alpha MAPK resulted in decrease in the uPA promoter activity in SNU-638 cells. These results suggest that P38 MAPK signaling pathway is important for uPA expression in gastric SNU-638 cells by enhancing the promoter activity of uPA. PMID- 12883726 TI - Follow-up CT findings of tamoxifen-induced non-alcoholic steatohepatitis (NASH) of breast cancer patients treated with bezafibrate. AB - One-third of the breast cancer patients who underwent tamoxifen intake showed less than 0.9 of their liver/spleen CT (computed tomography) ratio on their annual CT study, and were diagnosed as having fatty liver (hepatic steatosis). Among them, patients who showed a lower liver/spleen CT ratio of less than 0.5 were recommended to undergo needle biopsy of the liver in order to obtain histopathological confirmation of non-alcoholic steatohepatitis (NASH), with 15 patients undergoing needle biopsy of the liver. As a result, 14 out of the 15 patients were diagnosed as having NASH, and these patients were additionally administered bezafibrate in order to prevent possible progressive changes of NASH into liver cirrhosis. In this study, we show the changes of follow-up CT findings of 6 patients with histopathologically-proven NASH who continued to undergo bezafibrate intake after the diagnosis of NASH. Two patients showed almost complete improvement as indicated by the liver/spleen CT ratio several months after completion of a tamoxifen intake of 5 years, and another 3 showed partial improvement on their liver/spleen CT ratio by bezafibrate intake in spite of continuing tamoxifen intake. Another patient with diabetes mellitus (type II) showed a continually decreasing liver/spleen CT ratio during adjuvant tamoxifen in spite of bezafibrate intake. Therefore, we concluded that the progression of NASH could be prevented by bezafibrate without any interruption of adjuvant tamoxifen treatment. For patients with diabetes mellitus, critical follow-up using CT study and laboratory tests is considered essential. PMID- 12883727 TI - Protracted administration of weekly docetaxel in metastatic breast cancer. AB - The purpose of this study was to evaluate the efficacy and toxicity of weekly docetaxel in metastatic breast cancer. Fifty-seven patients were enrolled in this study of weekly docetaxel given at 25 mg/m(2)/week as a 1-h infusion in out patient setting. Each cycle consisted of 3 weeks of therapy followed by 1-week treatment break, and the patients received a median of 24 infusions with a median cumulative dose of 600 mg/m(2). Overall response rate was 30% [95% confidence interval (CI), 18-42%], and 31% (18-42%) of patients had stable disease for at least 6 months. With a median follow-up of 16 months, the median time to treatment failure was 11 months (6-14 M), and 3-year survival rate was 60%. There was no grade 4 toxicity, and myelosuppression, fatigue, nausea, vomiting was mild, thus the regimen was generally well tolerated. On the other hand, 44% of patients had grade 1-2 nail change, 21% had grade 3 pleural effusion, 17% had grade 1-2 tearing, and these became more common with prolonged administration of docetaxel. Thus, a weekly schedule of docetaxel at low dose maintains activity and is less myelosuppressive, however, one should be careful about cumulative toxicities. PMID- 12883728 TI - FDG PET in suspected recurrent and metastatic prostate cancer. AB - Development of an accurate non-invasive imaging technique to detect recurrent and metastatic prostate cancer is critical for the effective management of these patients. The purpose of our study was to determine the diagnostic utility of positron emission tomography with [F-18]fluorodeoxyglucose (FDG PET) in patients with suspected or known metastatic and recurrent prostate cancer. We performed 12 FDG PET scans in 12 men (age 65-81 years) with history of prostate cancer who had previously undergone radical prostatectomy (n=3) or prostate radiotherapy (n=9). Serum prostate specific antigen (PSA) level was elevated in all patients (5-206 ng/ml). Available correlative imaging studies included contrast-enhanced chest, abdomen and pelvis CT (n=8), bone scintigraphy (n=5), and radiography (n=2). PET findings were compared to the findings of the other imaging studies on a lesion by-lesion basis in individual patients. Validation was by clinical or imaging follow-up for up to 1 year. PET findings were concordant with the findings of the other imaging studies in 7 patients. PET was discordant with the other imaging studies in 5 patients. PET demonstrated suspicious hypermetabolic pelvic lymph nodes in one patient with negative pelvis CT. PET underestimated the extent of osseous metastatic disease in the remaining 4 patients. FDG PET is limited in the detection of osseous metastatic lesions but may be useful in the detection of metastatic nodal and soft tissue disease. PMID- 12883729 TI - Cisplatin-induced ubiquitination of RNA polymerase II large subunit and suppression of induction by 7-hydroxystaurosporine (UCN-01). AB - Exposure of cells to DNA-damaging agents induces hyperphosphorylation of the C terminal domain (CTD) of mammalian RNA polymerase II (RNAP II) large subunit (LS); the hyperphosphorylated RNAP II is then ubiquitinated. The purpose of this study was to verify that cisplatin-induced RNAP II ubiquitination is transcription dependent in living cells and to determine whether 7 hydroxystaurosporine (UCN-01) inhibits the ubiquitination induced by cisplatin. Cisplatin at clinically achievable concentrations (2.5-10 micro M) induced the ubiquitination of RNAP II in exponentially growing A2780 human ovarian tumor cells; the effect was drug-dose and exposure-time dependent. Such induction, however, was not observed in colcemid-selected mitotic cells. When detergent extraction was applied, the ubiquitinated RNAP II was recovered in the detergent insoluble fraction, indicating that the protein was tightly bound to DNA. In an in vitro transcription reaction that consists of nuclear extracts and an immobilized DNA template containing a site-specific cisplatin lesion, the elongating RNAP II that was stalled at a cisplatin lesion site on the template was targeted by ubiquitins. Together, our results indicate that the ubiquitination is associated with transcription-coupled repair. We previously showed that the Ser/The kinase-inhibitor UCN-01 inhibits nucleotide excision repair. Here, we further determined the effect of UCN-01 on the phosphorylation and ubiquitination of RNAP II LS in a whole-cell system. Immunoblotting results showed that UCN-01 suppressed the cisplatin-induced ubiquitination and the cisplatin-induced shift from the hypophosphorylated IIa to the hyperphosphorylated IIo, without affecting the basal levels of the IIo and IIa forms of the RNAP II CTD, suggesting that UCN-01 acts by suppressing cisplatin mediated induction of the one or more kinases that is responsible for the conversion of the IIo that is important for ubiquitination. PMID- 12883730 TI - Mutational analysis of ras gene family in lung cancer in Thai. AB - H-, K- and N-ras gene mutations were analyzed in lung cancer from Thai patients. Thirteen out of 58 cases (22%) harbored the mutations. Ten cases showed K-ras gene mutations at codon 12, 1 case presented a mutation at codon 13 and another case exhibited a mutation at codon 63. Silent mutations of N-ras gene in codons 57 and 62 were seen in one patient, whilst no H-ras mutation was found in these patients. Bases change in K-ras gene were G right curved arrow T transversion (62%), G right curved arrow A transition (15%) and G right curved arrow C transition (15%), whereas T right curved arrow G transversion and A right curved arrow G transition were detected in N-ras mutant gene. PMID- 12883731 TI - Role of the host microenvironment on the expression of VEGF-subtypes in squamous cell carcinoma. AB - The phenotypic characteristics and function of microvascular endothelium differs from tissue to tissue because of the organ specific microenvironment. Even the neovasculature in tumors differs depending upon tumor location. These findings indicate that local regulatory pathways modulate endothelial-cell growth and function in a tissue specific manner. This might be mediated by novel tissue specific regulators or the tissue specific regulation of known proteins. One of the major proangiogenic factors is VEGF. Its importance for tumor vascularisation and growth has been shown in several studies. VEGF is expressed in several subtypes and its impact on initial tumor growth and tumor progression has been discussed. Many of these results have been obtained by xenotransplantation studies in mice. The influence of the mouse microenvironment on the VEGF-subtype expression has not been previously analyzed. In the present study we analyzed the impact of the nude mouse microenvironment on the expression of the VEGF-subtypes after subcutaneous xenotransplantation. Our study was performed on seven squamous cell carcinoma (SCC) cell lines. We analyzed by quantitative PCR the mRNA expression levels and the proportion of the subtypes -121, -165, -189, and of VEGF-total of established xenotransplanted tumors and compared them to the cell lines prior to transplantation. We found high VEGF-165 fractions in the cell lines leading to fast growing, large tumors. The proportion of the VEGF-isoforms remaining in the cell lines and xenotransplanted tumors were generally unchanged, but an overall decrease of VEGF by about 50% was observed. We conclude that the VEGF-subtypes expressed by the transplanted tumors are not differentially regulated by the nude mouse subcutaneous microenvironment and that the tumor growth characteristics are not dependent on VEGF-subtype regulation of the host. PMID- 12883732 TI - Traditional Chinese medicine Astragalus reverses predominance of Th2 cytokines and their up-stream transcript factors in lung cancer patients. AB - Th2 cytokine is predominant in tumor patients and was found to be associated with tumor progression. Reversing of Th2 dominant status is thought to be a promising strategy. In the present study, peripheral blood mononuclear cells (PBMNC) of 37 lung cancer patients and 19 healthy subjects were prepared and used for examination of cytokine secretion and gene expression. The positive percentage of mRNA transcripts of Th1 cytokines (8.1% for IFNgamma and 13.5% for IL-2) in patients' PBMNC were lower than those of Th2 cytokines (70.3% for IL-4, 64.9% for IL-6 and 83.8% for IL-10). The gene expression capacity (measured as relative intensity to ratio of beta-actin) of patients for Th1 cytokines was low, but constitutively relatively high for Th2 cytokines. Both positive percentage and relative intensity were lower in transcript factor for Th1 cytokine, T-bet (40.5% and 0.139, respectively) than those for Th2 cytokine, GATA3 (89.2% and 0.364, respectively). Traditional Chinese medicine, Astragalus (AG) was observed to reverse Th2 status of lung cancer. AG enhanced culture supernatant and gene expression levels of Th1 cytokine (IFNgamma and IL-2) and its transcript factor (T-bet), and reduced those of Th2 cytokines in cultured PBMNC of lung cancer patients. These results demonstrated that traditional Chinese medicine AG might reverse the Th2 predominant status in lung cancer patients, which is a probable alternative therapeutic regime in future. PMID- 12883733 TI - Pentostatin in T-non-Hodgkin's lymphomas: efficacy and effect on CD26+ T lymphocytes. AB - Pentostatin is an adenosine deaminase (ADA) inhibitor with antineoplastic activity. CD26 is a surface glycoprotein with a key role in T cell function as the ADA binding protein. We conducted a phase II study to evaluate pentostatin efficacy in relapsed T-non-Hodgkin's lymphoma (T-NHL) and to correlate response with tumor CD26 expression. We also examined the lymphopenic effect of pentostatin on CD26+ T lymphocytes. Eighteen patients were registered for the study. Pentostatin was administered as intravenous bolus daily over 3 days at an initial dose of 5 mg/m(2)/day, repeated every 4 weeks. CD26 surface expression on tumor cells and T lymphocytes was determined by flow cytometry. Out of 14 patients evaluable for response, there was 1 (7%) complete response (CR) and 6 (43%) partial responses (PR). Median progression-free survival for responders was 6 months (range: 2-15 months); median number of courses was 4 (range: 1-6). Responders included 1 of 2 CD26+ and 5 of 9 CD26- cases. Pentostatin also specifically depleted CD26+ rather than CD26- T lymphocytes, potentially associated with immunosuppression. We therefore conclude that while pentostatin is a safe and active agent for T-NHL regardless of CD26 expression, it may selectively deplete CD26+ T lymphocytes, with potentially significant clinical implications. PMID- 12883734 TI - Aberrant CpG island methylation in neurofibromas and neurofibrosarcomas. AB - Aberrant methylation of the promoter CpG island of human genes is an alternative gene inactivation mechanism that contributes to the carcinogenesis of human tumours. We have determined the methylation status of the CpG island of 11 tumour related genes (RB1, p14ARF, p16INK4a, p73, TIMP-3, MGMT, DAPK, THBS1, caspase 8, TP53 and GSTP1) in 18 neurofibromas (including one plexiform neurofibroma) and three neurofibrosarcomas, as well as two non-neoplastic peripheral nerve sheath samples, using methylation-specific polymerase chain reaction. The series included sporadic and neurofibromatosis type 1-associated tumours. The incidence of aberrant methylation in the tumour samples was 52% for THBS1, 43% for MGMT, 33% for TIMP-3, 19% each for p16INK4a and p73, 14% for RB1, 5% for p14ARF, and 0% for DAPK, caspase 8, TP53 and GSTP1. No methylation of these genes was detected in the two samples of non-neoplastic peripheral nerve sheath. All but three samples in the study displayed aberrant methylation in at least one of the studied genes, and there was no correlation between methylation status and the patients' clinical parameters. These findings suggest that methylation of some tumour-related genes may play a significant role in the tumourigenesis of neurofibromas/neurofibrosarcomas. PMID- 12883735 TI - Modified radiosensitivity of pancreatic cancer xenografts by farnesyl protein transferase inhibitor and MEK inhibitor. AB - We investigated the effects of the farnesyl transferase inhibitor (FTI) manumycin and the MEK inhibitor PD98059 on growth of human pancreatic cancer, with mutant (SUIT2) or wild-type (BxPC-3) K-ras, xenografted into nude mice. Tumor growth was not reduced by either of the agents at a dose of 3 mg/kg without irradiation. Growth of SUIT2 irradiated at 15 Gy or 30 Gy was reduced by manumycin and PD98059: at 15 Gy, tumor volume doubling time (TVDT) increased from 18.6+/-3.8 to 36.3+/-14.2 days with PD98059 (p<0.05); at 30 Gy, TVDT increased from 32.8+/-6.8 to 70.5+/-10.5 days and 70.7+/-1.5 days, respectively. Manumycin tended to reduce growth of BxPC-3, but the difference in TVDT was not statistically significant. PD98059 significantly increased the TVDT of BxPC-3 at 30 Gy from 34.4+/-18 to 62.6+/-9.8 at 30 Gy. The present results suggest that Ras signaling pathways are potential targets for manipulation of radiosensitivity, and that induction of an alternative pathway may enhance radiosensitivity of pancreatic cancer. PMID- 12883736 TI - Effects of 1alpha,25-dihydroxy-vitamin D3 pretreatment and MAP kinase inhibitor PD 98059 on response of osteoblasts to prostate-derived osteoblastic factors. AB - Prostate carcinoma-derived factors induce a proliferative response in osteoblasts. The present study investigated the involvement of MAP kinase in the osteoblastic reaction of osteocytes and the response of 1alpha,25-hydroxy-vitamin D3 (1,25-vitD3)-pretreated osteoblasts. Conditioned media (CM) from prostate, colon, pancreatic, renal cell and breast cancer cell lines were tested on their proliferative activity using murine osteoblast-like MC3T3-E1 cells, MG63 human osteosarcoma cells and immortalized human osteoblasts (AHTO-7). Changes in osteoblastic activities of the supernantants were measured in the presence of MAP kinase inhibitors and following 1,25-vitD3-induced differentiation of the target osteoblasts. Supernatants of prostate cancer cells stimulated proliferation of osteoblasts in all three indicator cell lines, with AHTO-7 exhibiting the most significant correlation to human primary osteoblast cultures. 1,25-vitD3 induced the differentiation marker alkaline phosphatase (ALP) in MC3T3-E1 and AHTO-7, but only to a minor degree in MG63 cells. 1,25-vitD3-induced differentiation reduced the proliferative response to CM from several cell lines in MC3T3-E1 and MG63 to a minor degree, whereas in AHTO-7 cells the osteoblastic reaction was reduced for 2/4 pancreatic, 3/3 colon and 1/1 renal cancer CMs, however not for 3/3 prostate cancer CMs. Stimulation of AHTO-7 cells by CM from prostate cancer lines is inhibited significantly by MEK1 kinase inhibitor PD 98059 in contrast to CMs derived from other carcinomas, except ACHN renal cancer cells. The findings in the present study demonstrate that human AHTO-7 cells seem to represent a valid human system to monitor osteoblastic activity, especially in respect to 1,25 vitD3-induced differentiation. Vitamin D3-induced differentiation has no direct effect on prostate cancer-derived osteoblastic activity in the same cell line in vitro, which however, could be reversed by disruption of the signal transduction at the MAP kinase level, revealing a new target for the inhibition of prostate cancer-associated bone formation. PMID- 12883737 TI - High plasma YKL-40 level in patients with ovarian cancer stage III is related to shorter survival. AB - YKL-40 (human cartilage glycoprotein-39) is a member of family 18 glycosyl hydrolases. YKL-40 is a growth factor and is secreted by cancer cells. High serum levels of YKL-40 in patients with colorectal cancer and recurrent metastatic breast cancer have been associated with a poor prognosis. We evaluated the prognostic value of plasma YKL-40 in patients with primary ovarian cancer (OC). YKL-40 was determined by ELISA in plasma obtained preoperatively from 47 women with stage III OC and in plasma from 79 healthy females. The results showed that plasma YKL-40 was elevated compared to healthy females in 57% of the OC patients and was highest in the patients who died during the follow-up compared to the patients still alive (186 vs. 78 micro g/l, p=0.002). Patients with high plasma YKL-40 (>130 micro g/l) had significantly (p=0.0003) shorter survival than patients with normal plasma YKL-40. Multivariate Cox regression analysis showed that plasma YKL-40 (RH=3.95; 95% CI, 1.52-10.27; p=0.005) and radicality after primary surgery (RH=4.03; 95% CI, 1.81-8.97; p=0.001) were independent prognostic factors of survival, whereas age, histological type of tumour and serum CA125 had no independent prognostic value. In conclusion, plasma levels of YKL-40 proved of prognostic value in stage III OC patients. PMID- 12883738 TI - Expression and intracellular localization of Smad proteins in human endometrial cancer. AB - The aim of our study was to examine expression of Smad proteins i.e., Smad2, Smad3 and Smad4 both as mRNA and protein as well as their intracellular localization in normal (n=13) and neoplastic (n=42) endometrial tissue specimens using RT-PCR and immunological techniques i.e., Western blot and ELISA. Two uncommon female genital tract tumours, rhabdomyosarcoma of uterine of the cervix and uterine carcinosarcoma were also included. No statistically significant differences were found in the mRNA level of the examined Smad proteins between normal and tumour tissue specimens. Smad2 and Smad3 mRNAs were detected both in uterine carcinosarcoma and rhabdomyosarcoma of the uterine cervix. However, significantly lower Smad2 and Smad4 mRNA level was noted when the depth of myometrial invasion was considered (p<0.05). In endometrial cancer as compared to normal endometrium significantly higher levels of Smad2 and Smad3 proteins, both in cytoplasmic (p=0.002; p=0.0001) and nuclear (p=0.016; p=0.0004) fractions were observed. Both in uterine carcinosarcoma and rhabdomyosarcoma of the uterine cervix Smad2, Smad3 and Smad4 proteins were not detected. Moreover, significantly elevated Smad4 protein level in cytoplasmic fraction was stated when tumour grade and depth of myometrial invasion was undertaken (p<0.05). When intracellular distribution of Smads was considered differences between cytoplasmic and nuclear localization in normal and carcinomatous endometrium was stated. In endometrial cancer decreased number of cases with Smad3 and increased number of cases with Smad4 located in nuclear fraction was found. In conclusion, the disturbances in Smad protein expression and/or differences in their intracellular distribution suggest, that TGF-beta signaling pathway via Smads may be deregulated in endometrial carcinomas. PMID- 12883739 TI - Predicting factors for collection of peripheral blood stem cells in patients with advanced germ cell tumors after cisplatin-based combination chemotherapy. AB - The objectives of this study were to identify the predictive factors for the mobilization of CD34 positive peripheral blood stem cells (PBSC) and to determine the optimal timing of PBSC harvest in patients with advanced germ cell tumors after cisplatin-based combination chemotherapy. Thirty-eight patients with advanced germ cell tumors were enrolled in this study. We undertook a retrospective analysis of 124 aphereses performed between March, 1994 and December, 1999. The predictive value for the optimal timing of PBSC harvest was determined by the analysis of the correlation factors between the number of harvested CD34 positive cells and several clinicopathological factors. The mean number of CD34 positive cells obtained at a single apheresis was 5.38 x 10(6)/kg, and the mean cumulative number of CD34 positive cells in each patient was 24.07 x 10(6)/kg. The number of CD34 positive cells were significantly correlated to the number of previously performed chemotherapies before PBSC harvest, leukocyte count and the percentage of immature leukocytes (myelocytes plus metamyelocytes) (p=0.0098, p=0.011, and p<0.0001, respectively). Multivariate analysis revealed that the number of chemotherapies and the percentage of immature leukocytes were independent predictors for the number of harvested CD34 positive cells (p=0.012 and p=0.016, respectively). The present findings suggest that PBSC harvest should be performed during first-line chemotherapy, and that the monitoring of the percentage of immature leukocytes could be a useful predictor for determining the optimal timing of PBSC harvest. PMID- 12883740 TI - Rad52 gene mutations in breast/ovarian cancer families and sporadic ovarian carcinoma patients. AB - Rad52 encodes a protein which is required for recombinational repair of double strand breaks. It is also associated with breast cancer predisposition genes BRCA1 and BRCA2. Mutations in the genes Rad51 or Rad52 result in severe defects in genetic recombination and the repair of double-strand DNA breaks. In order to examine if Rad52 mutations might be involved in sporadic ovarian cancer, we analyzed two stop mutations (Ser346ter and Tyr415ter) in 142 Austrian ovarian carcinoma patients and 128 healthy volunteers. In addition, we analyzed these two mutations in 105 breast/ovarian cancer families (160 members) to examine if the mutations in Rad52 are associated with the occurrence of cancer and with mutations in the BRCA1 and BRCA2 genes. Our results show that these two mutations are rare in all three groups examined. There are no statistically significant differences in the frequencies of the Rad52 mutations between the control group and sporadic ovarian cancer patients and between the control groups and familial breast/ovarian cancer patients, indicating that these two mutations of the Rad52 do not play a major role in the initiation of sporadic ovarian carcinoma and familial breast/ovarian cancer. PMID- 12883741 TI - The alpha1-adrenoceptor antagonist terazosin induces prostate cancer cell death through a p53 and Rb independent pathway. AB - Prostate cancer is the second leading cause of cancer-related death in men. Treatment failure in prostate cancer is usually due to the development of androgen independence and resistance to chemotherapeutic drugs at an advanced stage. Recently, it was reported that the alpha1-adrenoceptor antagonist terazosin was able to inhibit prostate cancer cell growth and indicated that it may have an implication in the treatment of prostate cancer. The aim of the present study was to investigate the mechanisms involved in terazosin-induced prostate cancer cell death using two androgen-independent cell lines, PC-3 and DU145. Our results showed that terazosin inhibited not only prostate cancer cell growth but also colony forming ability, which is the main target of chemotherapy. We also found that the sensitivity of these cells to terazosin was not affected by the presence of either functional p53 or Rb, suggesting that the terazosin induced cell death was independent of p53 and Rb. However, the terazosin-induced cell death was associated with G1 phase cell cycle arrest and up-regulation of p27KIP1. In addition, up-regulation of Bax and down-regulation of Bcl-2 was also observed indicating that these two apoptotic regulators may play important roles in terazosin-mediated cell death pathway. Our results provide evidence for the first time that terazosin may have a therapeutic potential in the treatment of advanced prostate cancer. PMID- 12883742 TI - Molecular patterns of nuclear and mitochondrial microsatellite alterations in breast tumors. AB - The widespread distribution of microsatellite sequences within the human genome has allowed researchers to identify alternative patterns of microsatellite alterations in cancer cells. Among them, aneuploid patterns of nuclear microsatellites, pseudo-diploid microsatellite instability patterns, and also patterns of microsatellite instability within the mitochondrial genome. In this context, while aneuploid patterns of multiple genomic gains and losses had already been described in breast tumors, data on nuclear microsatellite instability still remain controversial and limited data on mitochondrial instability are available. In order to clarify this, we undertook an extensive analysis of nuclear and mitochondrial microsatellite alterations in breast ductal adenocarcinomas, stratified by grades. No instability was detected in any of the 40 dinucleotide microsatellites analysed nor in bat26 and APDelta3 mononucleotide repeats, clearly concluding that microsatellite instability is not a feature of ductal breast tumorigenesis. Instead, microsatellites defined a clear pattern of aneuploid genomic gains and losses among which, losses of BRCA1 at D17S855 and gains of plakoglobin at D17S846 significantly associated to grade III tumors and poor prognosis. On the other hand, mitochondrial instability at the transcription control region was also detected in 10.8% of cases. None of the new mitochondrial variants was found in the normal tissue counterparts, confirming that these new variants arise as sporadic somatic mutations in the tumor cells. Also, no association was found between heteroplasmy in the normal tissue and mitochondrial instability in the tumors. We therefore suggest that these new variants arise in tumors as a consequence of the progressive accumulation of slippage somatic mutations and the intrinsic instability of these microsatellite sequences. Finally, our results also confirm that mitochondrial instability does not associate with nuclear MSI. PMID- 12883743 TI - Establishment and characterization of sublines of LNCaP human prostate cancer cells. AB - The present study was undertaken to develop a panel of human prostate cancer cell sublines that represent a phenotypic continuum of prostate carcinogenesis. We cloned and established more than two dozen LNCaP sublines from parental LNCaP cells by the limiting dilution method, akin to a fluctuation analysis, in vitro. The newly established LNCaP sublines differ in hormone-sensitivity, anchorage independent growth ability and rate of PSA production. These LNCaP sublines may represent the naturally occurring heterogeneity in human prostate cancer and, therefore, could be useful for studying the effects of anticarcinogenic agents on cell clones that are derived from the same parental cell population. PMID- 12883744 TI - Biochemical characterization of soluble Tn glycoproteins from malignant effusions of patients with carcinomas. AB - The Tn determinant (GalNAc-O-Ser/Thr) is one of the most specific human tumor markers. In normal cells Tn is a cryptic structure in the peptide core of mucin type O-glycoproteins, and it is detected in an unmasked form in most human carcinomas evaluated by immunohistochemistry. Scarce data are available regarding the characteristics of soluble Tn bearing glycoproteins. We herein report the first comparative characterization of soluble Tn glycoproteins derived from different kinds of human tumors (breast, colon, gastric, ovarian and liver). Considerable heterogeneity was observed in the physicochemical properties of Tn soluble glycoproteins from all the tumor-associated effusions evaluated. In SDS PAGE analysis Tn glycoproteins from liver and colon effusions migrated as a broad single major component (>500 kDa), while several components of >200 kDa were identified in samples from breast, ovarian, and gastric cancer. The results of perchloric acid (PCA) treatment and CsCl gradient ultracentrifugation indicated that the Tn glycoproteins in effusion fluids correspond predominantly to mucin like glycoproteins. However, in samples from patients with colon and liver cancer, a fraction of Tn glycoproteins formed part of the immune complexes that precipitated in PCA, suggesting that the anti-Tn immune response in vivo could modify their physicochemical properties. The four apomucins evaluated (MUC1, MUC2, MUC5AC and MUC6) carried Tn epitopes in each of the effusions, indicating that soluble apomucin detection may reflect the abnormal expression of MUC genes inherent to these tumors. Taking together, these results indicate that apomucin expression profile is responsible, at least in part, for the high heterogeneity of soluble Tn glycoproteins, and suggest that the identification of Tn determinant on the different soluble apomucins could be useful for the development of new diagnostic tools as well as to evaluate the anti-tumor immune response in patients with cancer. PMID- 12883745 TI - Evaluation of anti-tumour effects of oral fenretinide (4-HPR) in rats with human neuroblastoma xenografts. AB - Neuroblastoma, the most common extracranial solid tumour in children, may undergo spontaneous differentiation or regression, but the majority of metastatic neuroblastomas have poor prognosis despite intensive treatment. Retinoic acid and its analogues regulate growth and differentiation of neuroblastoma cells in vitro, and 13-cis retinoic acid has shown activity against human neuroblastomas in vivo. Fenretinide [N-(4-hydroxyphenyl)retinamide] has been identified as a synthetic retinoid able to induce apoptosis of numerous malignant cell lines in vitro, including neuroblastoma. Furthermore, in animal models, fenretinide has shown chemopreventive and therapeutic efficacy against several malignancies without any obvious signs of toxicity. To investigate the anti-neuroblastoma tumour growth effects of oral fenretinide in vivo we used a human neuroblastoma xenograft model. Nude rats with established neuroblastoma xenograft tumours were treated orally with fenretinide for 10 days. Five different doses of fenretinide were used ranging from 2.5 to 75 mg/rat/day (10-300 mg/kg). Tumour volumes and toxic side effects were monitored during treatment and tumour weights were recorded at autopsy. In this study we found no significant anti-tumour growth effects of fenretinide in vivo, when used as oral treatment of rats with established neuroblastoma xenograft tumours. Furthermore, there were no intra tumoural differences in treated compared to untreated tumours. However, because of the promising results of fenretinide on neuroblastoma growth in vitro, further in vivo studies are warranted using other modalities of drug administration. PMID- 12883746 TI - Induction chemotherapy for bone sarcoma in adults: correlation of results with erbB-4 expression. AB - Tumor response to preoperative chemotherapy is an important prognostic factor for localized, operable extremity osteosarcoma. Other clinical variables include tumor size and location, age and sex, and serum enzymes. Advances in molecular oncology yielded a second group of factors such as multidrug resistance status, loss of heterozygosity of RB gene, and HER2/erbB-2 expression. The aim of this study was to investigate the expression and the prognostic value of the newly described erbB-4 receptor in specimens from adults with bone sarcomas treated by pre- and postoperative chemotherapy. Thirty-three patients with non-metastatic bone sarcoma have been treated by two doxorubicin-based induction chemotherapy regimen, followed by limb sparing surgery and tailored adjuvant chemotherapy. Pre chemotherapy tissue specimens were investigated for the expression of erbB-4 receptor and post-induction specimens were assessed for pathological response. The clinical response rates were 32-36%. The degree of induced necrosis was correlated with the disease-free survival (DFS). Patients achieving >/=90% necrosis had an improved DFS over patients with poor histological response. ErbB 4 expression was significantly associated with poor histologic response and shorter DFS. ErbB-4 expression may be used for prognostication of adults with bone sarcomas. PMID- 12883747 TI - Gleditsia sinensis fruit extract is a potential chemotherapeutic agent in chronic and acute myelogenous leukemia. AB - The anti-leukemia activity of the saponin rich Gleditsia sinensis Lam. fruit extract (GSE) was investigated on cancer cell lines and bone marrow cells obtained from consented patients with chronic myelogenous leukemia (CML) and acute myelogenous leukemia (AML) during presentation. The growth inhibitory activity of the extract was determined by [3-(4,5-dimethylthiazol-2-yl)-5-(3 carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium] (MTS) assay. Colony formation assay was performed to investigate the regeneration potential. Cellular morphology change was studied. Apoptosis was demonstrated by DNA electrophoresis, reverse transcription polymerase chain reaction (RT-PCR) and flow cytometry. The mean concentration to inhibit the cell growth by 50% (MTS50) was 18+/-1.6 micro g/ml for K562 CML cell line and 12+/-1.3 micro g/ml for HL-60 acute promyelocytic leukemia cell line. Patient samples showed a mean MTS50 of 13-28 micro g/ml. Non malignant hematological disorder bone marrow samples showed a mean MTS50 from 45 to 53 micro g/ml. Loss of regeneration property after treatment with GSE of these two cancer cell lines were confirmed by colony formation assay. GSE was able to induce cell shrinkage in K-562. DNA laddering was observed by incubating the leukemia cells with GSE. RT-PCR demonstrated that the pro-apoptic gene bax was induced while the anti-apoptic gene bcl-2 and cell cycle active gene PCNA were reduced. Flow cytometric analysis showed that the apoptotic effect of GSE on leukemia cell line was time- and dose-dependent. Thus GSE might be potentially used as a chemotherapeutic drug to treat patients with acute and chronic myelogenous leukemia. PMID- 12883748 TI - Variable cytotoxicity of amifostine in malignant and non-malignant cell lines. AB - Amifostine is the best known radioprotector and chemoprotector which has already been incorporated into general oncology practice. However, the data regarding its action at the cellular level remain unclear. The present study examined the effect of amifostine with and without ionizing radiation on the growth of malignant and non-malignant cell lines. Amifostine was found to have a remarkable cytotoxic effect on malignant epithelial cell lines but a modest cytotoxic effect on malignant melanoma and non-malignant cell lines. It demonstrated an additive effect with radiation therapy on the malignant cell line and a variable effect on the non-malignant cell line. Endothelial cells were not affected by amifostine, but the myoblast cells showed a synergistic effect of amifostine and radiation. These findings demonstrate that the cytotoxic as well as the radioprotective effect of amifostine are cell-specific. Thus, caution should be exercised in the use of amifostine as a radioprotector, and it should be tested for each model of disease. PMID- 12883749 TI - Genetic polymorphisms and susceptibility to esophageal cancer among Chinese population (review). AB - Esophageal cancer, which is prevalent in China and some other parts of the world, is a complex disease likely resulting from polymorphisms of multiple interacting genes and gene-environment interactions. Recent efforts have been made to analyze the associations between risk of this cancer and hereditary sequence variations in genes involved in metabolism, DNA repair and cell cycle control. We summarized here the results of published case-control studies that have examined the effects of common alleles of 15 genes, MTHFR, CYP1A1, CYP2A6, CYP2E1, GSTM1, GSTT1, GSTP1, NAT2, XRCC1, XPD, hOGG1, MGMT, p53, CNDD1 and L-Myc, on risk of esophageal squamous cell carcinoma among Chinese. Statistically significant differences in genotype frequencies found in case-control comparisons were MTHFR C677T and A1298C polymorphisms, the XRCC1 Arg194Trp polymorphisms, the hOGG1 Ser326Cys polymorphism, and the p53 Arg72Pro polymorphism. The overall effects of these genetic polymorphisms were moderate in terms of relative risk, with ORs ranging from 2-10. There was also some evidence that genetic polymorphisms in certain carcinogen-metabolizing enzymes such as CYP2E1, CYP1A1, CYP2A6, GSTM1, and GSTP1 modulate risk of the cancer, although the results require confirmation with larger sample size studies. For polymorphisms in GSTT1, XPD, CCND1, and L-Myc, the risk estimate from the studies was sufficiently precise to exclude an OR >/=1.5. PMID- 12883750 TI - Angiogenesis and the immune response as targets for the prevention and treatment of colorectal cancer (review). AB - The carcinogenic process in previously normal human colonic mucosa involves hyperproliferation and adenoma formation, but it is not known why only a tiny proportion of adenomas undergo malignant transformation. There is accumulating evidence in favour of the hypothesis that inflammatory processes are a prerequisite for the development of malignancy. The data include upregulation of mediators of the inflammatory response such as cyclooxygenase-2, the generation of inflammatory cytokines which result in induction of cell proliferation and inhibition of apoptosis, and chronic inflammation which may lead to the production of reactive species that damage DNA. Angiogenesis, the formation of new blood vessels from an existing vasculature, is generally regarded as essential to the late stages of carcinogenesis, allowing tumours to grow beyond 1 2 mm in diameter, invade and metastasise. In this communication, it is argued that angiogenesis is not only present before neoplastic transformation occurs, but that it is of relevance to inflammatory diseases that increase risk of developing colorectal cancer. It is proposed that intervention to prevent or treat colorectal cancer should be targeted at inhibiting inflammation, reducing angiogenesis and stimulating cell mediated immune responses. PMID- 12883751 TI - The Chinese medicinal herbal formula ZYD88 inhibits cell growth and promotes cell apoptosis in prostatic tumor cells. AB - Prostate cancer is a leading cause of cancer death in American males. Currently, there is no curative therapy available once prostate cancer has metastasized. A major systemic therapy for metastatic prostate cancer is anti-androgen therapy. Unfortunately this therapy is only palliative and rarely curative, and eventually the tumor cells develop resistance to further hormone manipulation. It is therefore imperative to develop alternative effective therapies. In the present study, the effect of a Chinese herbal formula, ZYD88, on regulation of cell growth and cell apoptosis was examined in prostatic tumor cells. ZYD88 decreased cell viability of multiple prostatic tumor cell lines, DU-145, PC-3, MDA-PCa 2b and LNCaP in a time- and dose-dependent manner. It also produced a rapid and dose dependent increase in caspase 3 activity in LNCaP and PC-3 cells, and induced DNA fragmentation in LNCaP cells, indicating cell apoptosis. In cotransfection assays, ZYD88 inhibited androgen-induced prostate specific antigen (PSA) gene promoter activity, and induced estrogen-target gene promoter activity. These data suggest that ZYD88 is a potential agent for prostate cancer therapy, and deserves further study. PMID- 12883752 TI - Evaluation of immunohistochemical parameters as prognostic markers in malignant fibrous histiocytoma. AB - In this study 65 primary malignant fibrous histiocytomas (MFH) of high malignancy grade were characterized by immunohistochemistry for their expression of proteins reflecting or promoting tumor growth. The results were evaluated in relation to the disease-free survival and the occurrence of metastases alone or in combination with local recurrences during follow-up. A tumor size >8 cm was strongly associated with both a shorter disease-free survival (p=0.001) and a higher frequency of metastases alone or together with local recurrence during follow-up (p=0.001 and 0.004). Similarly a higher frequency of mitosis was associated with a shorter disease-free survival (p=0.004), while the presence of necrosis or malignancy grade 4 did not affect the clinical outcome. No significant effect on the clinical outcome was seen for p53, Ki-67, p27 expression or for vascular density determined by factor VIII staining. However, a significant association was demonstrated between high Bcl2 expression and the risk to develop both local recurrence and metastases (p=0.026). Taken together, the findings support the importance of the tumor size, and suggest that bcl2 staining but not p53, Ki-67, p27, vascular density or distinction of grade 3 and grade 4 tumors are of clinical value in the prognostication of MFH tumors. PMID- 12883754 TI - [The superior labrum biceps tendon complex. Bringing more light into the darkness]. PMID- 12883753 TI - GM-CSF secreted by murine adenocarcinoma cells modulates tumor progression and immune activity. AB - During tumor development, growth factors may act in autocrine manner stimulating cell proliferation, or in paracrine manner affecting the microenvironment of the tumor and modulating the immune system. Murine mammary adenocarcinoma M3 tumor bearers develop lung metastases and leukocytosis during its evolution. Previously we described that M3 conditioned media enhanced metastasis incidence, when it was inoculated in tumor-operated mice. In the present study we determine that spleen cells from M3 tumor operated mice treated with M3 conditioned media, were able to transfer the capacity to enhance metastasis to other tumor operated mice. Spleen cells have immune suppressor activity that could be reversed by cyclophosfamide treatment. M3 tumor cells secrete GM-CSF, which is able to promote in vitro proliferation of M3 cells as well as spleen cells. This proliferation could be abrogated by the addition of anti-GM-CSF. We report that the GM-CSF secreted by M3 tumor cells had stimulatory activity on M3 tumor cell and lymphocyte proliferation. PMID- 12883755 TI - [Anatomy of the superior labrum complex of the shoulder]. AB - The outstanding progress achieved in MRI and arthroscopic techniques has forced orthopedic surgeons to consider the anatomy of the shoulder joint in detail. The superior labrum complex shows a broad range of variations, which is the reason for some difficulties regarding diagnostic conclusions as well as therapeutic considerations. In particular, the differentiation between the normal and pathological adherence of the labrum is a serious clinical problem. The glenoid labrum consists of circularly arranged collagenous fibers attached to the osseous margin of the glenoid by a fibrocartilaginous transitional zone. The cranial part of the labrum is more compact than the caudal one. It is also connected to the joint capsule and the integrated ligaments. Most of the fibers of the long head of the biceps tendon insert at the supraglenoid tubercle, only some of them continue in the fibrous ring. So-called SLAP lesions have to be differentiated from variations. In up to 18% of the patients normal clefts can be seen, e. g., sublabral holes, and in less than 6% other rare variations, e. g., the so-called Buford complex. PMID- 12883756 TI - [Anatomy of the glenoid labrum]. AB - Most instabilities or pain syndromes are associated with injuries or morphologic changes in the glenoid labrum complex or long head of the biceps tendon origin. The first anatomic descriptions go back to Fick in 1910 and since then many authors have described the anatomy of these structures. It was Snyder who introduced the term SLAP lesions, classifying superior, anterior, posterior labrum changes into four grades. It is still unclear whether all of the described and arthroscopically observed changes are due to a post-traumatic, acquired lesion or whether anatomic variations can be present as well. In order to elucidate this problem, 36 cadaver shoulder joints were inspected macroscopically and sectioned for microscopic evaluation. Here the glenoid could be divided into an superior and an anterior- superior area demonstrating a wide variety of morphologic labral glenoid changes, while the dorsal and inferior sectors of the glenoid showed a relatively uniform anatomy of a firm labrum-glenoid bond. Four types of biceps tendon attachments could be identified similar to the description given by Vangsness. In addition, a variety of anterior-superior changes could be found. The sublabral hole as described by Esch in the clinical setting was found to be a physiologic variant. Precise knowledge of the anatomic morphology of the normal glenoid in its variations seems to be necessary to understand variants and allow for distinguishing between physiologic anatomic variants and pathoanatomic changes in imaging and the clinical setting. PMID- 12883757 TI - [Normal anatomical variants of the superior labrum biceps tendon anchor complex. Anatomical and magnetic resonance findings]. AB - There are difficulties in diagnosing pathologies of the labrum-biceps tendon complex (LBTC) because of the great individual variability of this structure. Anatomical variations, such as the sublabral recess, are frequently found and can also be misinterpreted as Andrews or SLAP II lesions. The etiology and mechanisms of creating such a sublabral recess are not exactly known. The examination of 31 cadaveric shoulder specimens from an age group between 49.3 and 83.6 years showed a sublabral recess in 63.6-85%. This high frequency of sublabral detachment of the labrum in older patients indicates that according to the high range of motion of the humeral head and therefore the changing angle of the long biceps tendon (LBS) a certain mobility of the superior LBTC is physiological and should not necessarily be seen as a sign of instability. Besides mobility-increasing factors such as overhead professions and sports with high repetitive maximal abduction and external rotation and the individual age, the type of insertion of the LBT at the glenoid influences the development of sublabral recess. Accordingly, posterior oriented insertion types of the LBT determine deeper and more posterior oriented sublabral recesses whereas in cases of anterior directed types of insertions no or less deep recesses can be found. Anamnestic aspects, such as the mechanism of injury and overhead activities, these macroscopic and functional associations in MR arthrography and arthroscopic inspection have to be considered in order to differentiate between physiological and traumatic changes of the labrum-biceps tendon complex. PMID- 12883758 TI - [MR imaging of variants of the superior labral-bicipital complex and SLAP lesions]. AB - In the detection of SLAP (superior labral anterior to posterior) lesions of the shoulder MR arthrography shows a significantly higher sensitivity compared with conventional MR techniques and therefore, represents the method of choice in diagnostic imaging of the superior labral-bicipital complex. On the basis of morphological criteria it mostly allows distinction of traumatic lesions and anatomic variants of the superior labrum and the biceps anchor as well as classification of detected SLAP lesions (types 1-4 according to Snyder). However, the differentiation of a SLAP type 2 lesion and a sublabral recess can be very difficult, even if all distinction criteria are considered. PMID- 12883759 TI - [Biomechanical tests for type II SLAP lesions of the shoulder joint before and after arthroscopic repair]. AB - Superior labral anterior-to-posterior (SLAP) lesions can cause shoulder pain partly by causing glenohumeral instability. The purpose of this study was to examine the effect of a simulated type II SLAP lesion and subsequent repair on glenohumeral translation of the vented shoulder. In eight cadaver joints, a robotic/UFS testing system was used to measure joint translation by applying an anterior, posterior, or inferior load of 50 N to each shoulder. The "apprehension tests" for anterior and posterior instability were simulated by applying an anterior load of 50 N with an external rotation torque of 3 Nm or a posterior load of 50 N with an internal rotation torque of 3 Nm. Each loading condition was applied at 30 degrees and 60 degrees of glenohumeral abduction with a constant joint compressive load (44 N) to the intact, simulated SLAP lesion, and repaired shoulder. Repair of the type II SLAP was then performed by placing a Suretac through the labrum both anterior and posterior to the biceps anchor and testing was repeated. ANOVA was used to compare translation of the intact joint, the joint after the type II SLAP lesion had been simulated, and after repair. At 30 degrees of abduction, anterior translation of the intact vented shoulder joint from anterior loading was 18.7+/-8.5 mm and increased to 26.2+/-6.5 mm after simulation of the type II SLAP lesion ( p< or =0.05). The arthroscopic repair did not restore anterior translation (23.9+/-8.6 mm) to the same degree as the intact joint ( p> or =0.05). At 60 degrees of abduction, anterior translation of 16.6+/ 9.6mm in the intact joint was not significantly increased at 19.4+/-10.1 after simulation of the type II SLAP lesion ( p=0.0527). AP loading also resulted in inferior translation. At 30 degrees of abduction it was 3.8+/-4.0 mm in the intact joint and increased to 8.5+/-5.4 mm after the type II SLAP lesion ( p< or =0.05. After repair the inferior translation decreased significantly to 6.7+/-5.3 mm ( p< or =0.05). Although inferior translations were less at 60 degrees of abduction, results were similar to those at 30 degrees after repair. There were no significant increases in translation after SI/AP combined external rotation torque or posterior-anterior combined internal rotation torque loading. In this study the repair of a type II SLAP lesion only partially restored translations to the same degree as an intact vented shoulder joint. Therefore, improved repair techniques or an anteroinferior capsulolabral procedure in addition to the type II SLAP lesion repair might be needed to restore normal joint function. PMID- 12883760 TI - [Biomechanical investigations for the development of a SLAP-II-lesion]. AB - The superior labral-biceps-tendon-complex forms an anatomical and functional unit and combines static and dynamic elements of shoulder stability. At present, only theoretical hypotheses exist on the etiology of the microtraumatic SLAP-II lesion. To gain further insight into this, an instrument was developed to simulate throwing motions such as the late-cocking/early acceleration phase as well as deceleration/follow-through. Sixteen freshly frozen shoulder specimens were tested, varying the loads on the biceps tendon (25 N, 50 N, 100 N) and the compression of the humeral head against the glenoid (25 N, 50 N, 80 N). Each shoulder had to run through a certain number of cycles during the particular phase of throwing. The tests were stopped after a SLAP-II-lesion was observed, or after a limit of 15,000 cycles. Every 1,000 cycles the results of the tests were checked arthroscopically. A SLAP-II-lesion developed in only 10% of the specimens during the acceleration/ late cocking phase whereas in the deceleration/ follow through phase 83% developed such a lesion. According to our results, the deceleration/follow-through of the throwing motion seems to be responsible for creating microtraumatic SLAP-II-lesions. One reason is the loss of the centering function of the long head of the biceps tendon during total internal rotation, another is the increased posterosuperior translation of the humeral head in this position, which leads to a non-physiological contact, creating lesions in this area due to the large sheering forces. PMID- 12883761 TI - [Superior labrum-biceps anchor complex]. AB - The focus in overhead sport injuries of the shoulder has been directed on lesions of the superior labrum-biceps complex during the last few years. This is due to improved imaging modalities as well as a better understanding of the anatomy and biomechanics. Arthroscopy is capable of diagnosing and treating these lesions. The anatomy, biomechanics, and operative treatment strategies and techniques are described in detail. The superior labrum anterior-posterior (SLAP) lesion is found in young active overhead athletes. Based on their classification and concomitant lesions, the unstable SLAP lesions (types II and IV) have to be repaired in order to restore stability to the superior labrum-biceps complex for stabilizing the glenohumeral joint. According to our own results and after review of the current literature, SLAP repair yields good results in overhead athletes enabling them to return to pain-free participation in their sport. PMID- 12883762 TI - [The arthroscopic SLAP refixation]. AB - Lesions of the superior glenoid labrum and the insertion of the biceps tendon are a common cause for shoulder pain in patients performing overhead sports. The therapeutic management depends on the type of lesion, and should be carried out using an arthroscopic procedure. While type I SLAP lesions should be treated conservatively or with simple debridement, SLAP II, IV, and V lesions, with a detachment of the labrumanchor- complex, should be refixed with suture anchors. Only in cases of type III lesions with a bucket handle-like lesion of the labrum, but stable insertion of the biceps tendon, a simple debridement can be performed. From the biomechanical point of view, large type III lesions should also be reconstructed. To improve the arthroscopic view for preparation of the glenoid neck, an intraarticular loop ("Imhoff-suspension sling") can prevent the posterosuperior labrum from falling into the joint. The arthroscopic SLAP refixation is a technically highly demanding procedure which provides good clinical results for the patient. PMID- 12883763 TI - [SLAP-lesions of the shoulder]. AB - The advent of shoulder arthroscopy, as well as our improved understanding of shoulder anatomy and biomechanics, has led to the identification of previously undiagnosed lesions involving the superior labrum and biceps tendon anchor. Although the history and physical examinations as well as improved imaging modalities (i. e. magnetic resonance arthrography) are extremely important in understanding the pathology, the definitive diagnosis of SLAP lesions ("superior labrum anterior and posterior") is accomplished through diagnostic arthroscopy. Treatment of these lesions is directed according to the type of SLAP lesion. In general, type I and III SLAP lesions are debrided, whereas type II and many type IV lesions are repaired. The purpose of this article is to review the anatomy, classification, diagnosis and current treatment recommendations. PMID- 12883764 TI - [Results of repair of SLAP lesion]. AB - The surgical repair of SLAP lesions has become increasingly more common as the techniques and instrumentation have improved. However, there are few studies examining the results of repair of SLAP lesions. The goals of this paper are to summarize the recommended treatment for SLAP lesions and to report upon the studies available to date which report the results of SLAP repair. PMID- 12883765 TI - [SLAP lesions as a cause of posterior instability]. AB - SLAP lesions disrupt the perilabral architecture, but so far there have been no reports about posterior instability due to SLAP lesions. In a prospective study of 30 patients with recurrent posterior instability, we found SLAP lesions as a reason for instability in three cases. The purpose of this study is to point out that SLAP lesions can be a cause of posterior instability. Thirty patients with clinical posterior shoulder instability underwent diagnostic arthroscopy before operative stabilization procedures, three of whom (three males, aged 29-51 years) showed a SLAP lesion (once case each of types II, III, and IV) as a cause of posterior instability. All three patients had a history of a fall on the outstretched arm. All patients underwent arthroscopic refixation of the labrum. After arthroscopic refixation of the SLAP lesions, two patients were completely stable (SLAP II and III), whereas one patient (SLAP IV) reported microinstability during overhead activity but complete stability during activity of daily living. The same patient complained about moderate pain in extreme external-flexion rotation with slightly reduced range of motion in external-flexion position. All other patients were free of pain and showed free range of motion. Our results demonstrate that SLAP lesions can be a cause for posterior shoulder instability. In our cases, posterior shoulder instability caused by SLAP lesions was successfully treated by arthroscopic refixation of the torn biceps anchor. When treating posterior shoulder instability, SLAP lesions should be taken into account. PMID- 12883766 TI - [Follow-up treatment with physiotherapy after arthroscopic reconstruction in SLAP lesions]. AB - An appropriate physiotherapeutic treatment program complements an optimal operative result. In the instance of SLAP lesions, a differentiated therapy is only possible on consultation with the surgeon. A detailed treatment design allows optimal guidance of the patient to his or her personal goal. Important treatment elements include the optimization of the ability of the humerus head to centralize and the proprioception of the shoulder joint, as well as reinstatement of strength and endurance, especially of the rotator cuff. PMID- 12883767 TI - [Open versus arthroscopic shoulder stabilization. An analysis of the treatment costs]. AB - In this study the total costs of clinical open and arthroscopic anterior shoulder stabilization were evaluated, analyzed and compared. From 1988 to 1998 147 patients underwent open (Bankart) or arthroscopic (ASK) anterior shoulder stabilization. We randomized two groups of 30 patients for each method (Bankart: 25 male, 5 female, 29 years of age; ASK: 25 male, 5 female, 26 years of age) and evaluated the costs of their clinical treatment. The total cost was significantly ( p<0.05, Mann-Whitney U-Test) higher for the open (5639 euro) than for the arthroscopic (4601 euro) therapy. There was a significant difference between the groups for the average cost of surgery (Bankart: 2741 euro; ASK: 2315 euro, p<0.05) and the average postoperative treatment cost (Bankart: 2202 euro; ASK: 1630 euro, p<0.05) whereas the average preoperative treatment cost was not significantly different (Bankart: 669 euro, ASK: 657 euro). The staff costs for the surgical procedure (Bankart: 1800 euro (32%), ASK: 1319 euro (29%)) and the postoperative staff costs of the nurses (Bankart: 1271 euro (23%), ASK: 997 euro (22%)) represented the greatest parts of the total costs. The average duration of the clinical treatment was 15.8 days for the open- and 12,4 days for the arthroscopic group. PMID- 12883768 TI - [Advances in orthopaedic foot surgery. 9th Annual Congress of the German Association for Orthopaedic Foot Surgery, Jena, 14-15.03.2003]. PMID- 12883769 TI - [Arthroscopy of the talocrural articulation]. AB - Arthroscopy of the ankle joint meanwhile enjoys widespread use in those departments that perform arthroscopy so that the application of arthroscopic techniques can be considered a standard procedure also for the talocrural articulation. Various indications have become generally accepted and further areas of application will emerge. As is the case for the knee joint, arthroscopy of the ankle joint solely for diagnostic purposes has become obsolete and due to improved preoperative methods for diagnosis is no longer necessary as the sole procedure. The diagnostic work-up is normally followed by arthroscopic or open treatment. The multitude of clinical examinations and imaging techniques currently available enables noninvasive clarification of pathological conditions in the ankle joint to the greatest possible extent, making detailed preoperative planning possible in most cases. The present article describes the technique, indications, complications, and perspectives for arthroscopy of the talocrural articulation. PMID- 12883770 TI - Decomposition of buried corpses, with special reference to the formation of adipocere. AB - The unrestrained decomposition of a corpse involves the consecutive processes of autolysis, putrefaction and decay. Ideally, decomposition is completed within the regular resting time (15-25 years) and leads to the entire skeletalisation of the corpse. Adipocere, a greyish fatty substance formed during decomposition, is regarded as a spontaneous inhibition of post-mortem changes; it makes the corpse almost entirely resistant to decomposition and makes it impossible to use the same graves again. This creates problems for local governments with regard to the generally growing demand for burial ground. Apart from corpse-specific characteristics (e.g. sex, age, physique, cause of death), method of burial (e.g. material of the coffin, depth of grave, individual or mass grave, clothing) and time of burial, the conditions of the resting place (geology, topography, soil properties and frequency of use, air, water, and heat budget), in particular, have a special impact on adipocere formation. This study summarises the present knowledge on this phenomenon, combining results gained both in forensic medicine and in geosciences. PMID- 12883771 TI - Benzoquinones from millipedes deter mosquitoes and elicit self-anointing in capuchin monkeys (Cebus spp.). AB - Neotropical monkeys of the genus Cebus anoint themselves by rubbing arthropods and plants against their pelage. A recent study has shown that free-ranging wedge capped capuchin monkeys (C. olivaceus) in Venezuela self-anoint with a benzoquinone-secreting millipede, an activity by which they are hypothesized to appropriate chemical deterrents of mosquitoes. To evaluate the plausibility of this hypothesis, female yellow fever mosquitoes (Aedes aegypti) were presented with two millipede secretory compounds, 2-methyl-1,4-benzoquinone and 2-methoxy-3 methyl-1,4-benzoquinone, on nylon-reinforced silicone membranes placed over wells filled with human blood, a highly preferred food. Mosquitoes exhibited fewer landings, fed less frequently, and flew more frequently (a possible indication of repellency) in the presence of membranes treated with benzoquinones than with controls. These compounds also elicit self-anointing in captive male and female tufted (C. apella) and white-faced (C. capucinus) capuchin monkeys. PMID- 12883772 TI - Hot-blooded singers: endothermy facilitates crepuscular signaling in African platypleurine cicadas (Hemiptera: Cicadidae: Platypleura spp.). AB - The cicada genus Platypleura has a wide distribution across Africa and southern Asia. We describe endothermic thermoregulation in four South African species that show crepuscular signaling behavior. This is the first evidence of thermoregulation in platypleurine cicadas. Field measurements of body temperature ( T(b)) show that these animals regulate T(b) through endogenous heat production. Maximum T(b) measured was 22.1 degrees C above ambient temperature during calling activity at dusk. The mean T(b) during dusk activity did not differ from the mean T(b) during diurnal activity. A unique behavior for cicadas, a temperature dependent telescoping pulsation of the abdomen, was observed in the laboratory during endogenous warm-up. This behavior is part of a unique method of heat generation in endothermic cicadas. Males generally call from trunks and branches within the canopy and appear to use endothermy even when the sun is available to elevate T(b). Endothermy may provide the cicadas with the advantage of decreasing predation and acoustic competition by permitting calling from perches that most complement their cryptic coloration patterns and that ectotherms cannot use due to thermal constraints. In addition, endothermy may permit calling activity during crepuscular hours when atmospheric conditions are optimal for acoustic communication and predation risks are minimal. PMID- 12883773 TI - A longitudinal study of age-specific reproductive output and body condition among male rhesus macaques, Macaca mulatta. AB - In many mammalian species, male reproductive success appears to climb sharply at young adulthood, form a brief plateau during prime ages, and decline among older animals, a pattern often attributed to reduced physical condition with ageing. However, solid evidence to either substantiate or refute this profile among nonhuman primates is lacking. Here, we combine a decade of genetic analysis of paternity among free-ranging rhesus macaques, Macaca mulatta, with information about body condition in order to evaluate how changes in morphology might govern age-specific reproduction among males. We show that age-specific reproductive success traverses the same life history profile as found in other mammals, but reductions in reproductive output with advanced age were associated with reduced chances of survivorship rather than accompanied by diminished body condition. We demonstrate that variance in male age at onset of reproduction is three times greater than variance in female age at onset of reproduction. We provide the first evidence from primates that age-specific reproductive output among males is not a consequence of age-related changes in body condition, but reflects social and demographic factors. PMID- 12883774 TI - Synthesis and structure-activity relationship of diene modified analogs of Matsucoccus sex pheromones. AB - The biological activity of the Matsucoccus spp. sex pheromones and diene modified analogs has been tested in forests of Israel and Portugal in order to explore the structure-activity relationship of the pine bast scale pheromone/kairomone system. The response of the adult predatory bugs, Elatophilus hebraicus and E. crassicornis and of the brown lacewing, Hemerobius stigma is more selective than that of the conspecific Matsucoccus males. The removal of the terminal methyl group from the diene terminus of both pheromones 1 and 2 eliminates all kairomonal activity but retains moderate pheromonal activity. Addition of a methyl group to the diene terminus of pheromones 1 and 2 sustains full pheromonal and kairomonal activities of the Elatophilus spp. but eliminates entirely the kairomonal activity of H. stigma. Subtle designed alterations in the structure of the diene group, typical of all Matsucoccus pheromones, change the mode of the kairomonal activity markedly. PMID- 12883775 TI - Sumo wrestling in ants: major workers fight over male production in Acanthomyrmex ferox. AB - In the myrmicine ant Acanthomyrmex ferox, major workers have the same number of ovarioles as queens, thrice that of minor workers, making them well suited for egg-laying. In the queen's presence, infrequent aggression allows ranking of majors but they lay only unviable trophic eggs. Major workers engage each other, but not the minors, in antennal boxing and spectacular shaking contests, a novel interaction in ants. The absence of reversals allows a clear ranking of major workers. After queen removal, aggression becomes very frequent, but previous ranking is maintained. All majors start laying reproductive eggs although they show a skew in ovary development according to ranking. The dominant major, however, actively patrols the egg-pile and cannibalises eggs laid by subordinates. PMID- 12883776 TI - Is microhabitat segregation between two cicada species (Tibicina haematodes and Cicada orni) due to calling song propagation constraints? AB - The cicada species Tibicina haematodes and Cicada orni are two sympatric species often inhabiting vineyards. We show that they occupy two distinct levels: males of T. haematodes produce their calling songs from a high position in vine foliage while males of C. orni call from a low position near the ground on vine trunks. Experiments consisting of broadcasting and re-recording experimental signals in natural habitats from low and high positions show that signals are more and more modified as sender-receiver distance increases. T. haematodes would have an advantage when calling on trunks rather than on branches whereas C. orni would be able to call indiscriminately from both low and high positions. Thus, the microhabitat segregation observed between T. haematodes and C orni in vineyards does not seem to be related to calling song propagation constraints, but may be due to other ethological or ecological factors. PMID- 12883777 TI - An example of the prisoner's dilemma in biochemistry. AB - Two strains of microorganisms that both use sugar as energy resource, but which may choose between two different pathways of ATP production, are studied from a game-theory point of view. We consider these pathways as distinct strategies to which we assign payoffs that are proportional to the expected steady-state number of individuals sustainable on the basis of these strategies. In a certain parameter range, we find that the payoffs fulfil the conditions for the prisoner's dilemma. Therefore, cooperative behaviour is unlikely to occur, unless additional factors intervene. In fact, the yeast Saccharomyces cerevisiae uses a competitive strategy by fermenting sugars even under aerobic conditions, thus wasting its own resource. The simple quantifiable structure of the model should enable access to an experimentally determined payoff matrix. PMID- 12883778 TI - The emergence of collective foraging in the arboreal Gnamptogenys menadensis (Hymenoptera: Formicidae). AB - Gnamptogenys menadensis is an arboreal nester that forages opportunistically almost exclusively on vegetation, sometimes recruiting others to participate in prey retrieval. The three-dimensional characteristics of vegetation suggest that functions describing recruitment decision thresholds or the pattern of recruitment in arboreal species may differ from those predicted by optimal foraging theory. To examine the effects of prey abundance and distance on the recruitment dynamics of G. menadensis, we baited nests with one termite, five termites or a number of termites between 20 and 40 either near to or far from the entrance and observed the ensuing behaviors. G. menadensis recruited others when encountering multiple termites regardless of the termite pile's distance from the nest, although a few individuals remained at the site and defended the resource. The pattern of arrivals at the site indicates that the majority and sometimes all arrivals were recruited from the branch trails. In combination, these results suggest that the architecture of the foraging habitat, which limits available return routes to the nest and thus increases encounter probabilities with potential recruits, shaped the process of information transfer and generated a collective pattern of foraging and prey retrieval. PMID- 12883779 TI - [Diagnosis and therapy of osteoporosis. Strategy for effective treatment after fragility fractures]. AB - Until about 20 years ago osteoporosis was considered an inevitable disease of ageing. Now the situation has completely changed and osteoporosis is classified as a disorder which is easily diagnosed and treated in the early stages of its development that is,before a skeletal fracture has occurred. Two types of medication are available today for prophylaxis and therapy of osteoporosis: the antiresorptive and the osteoanabolic drugs whose efficacy has been demonstrated in large randomized clinical trials (RCTs). These drugs are effective not only in the early stages of osteoporosis i.e. for prevention, but also when fractures have already occurred, to reduce the risk of further skeletal fractures. However, in this setting only about 7% of patients in Germany are being treated although all are at significant risk of sustaining additional fractures. Consequently all patients with osteoporosis-related fractures should be thoroughly investigated during their hospitalisation and effective treatment instituted. In addition, treatment should be continued and monitored by the family doctor. The strategy for the administration of therapy for successful prevention of secondary fractures in osteoporosis is presented. PMID- 12883780 TI - [Mortality, morbidity and functional outcome after open book and lateral compression lesions of the pelvic ring. A retrospective analysis of 100 type B pelvic ring lesions according to Tile's classification]. AB - A retrospective study was made of 100 Tile type B and 122 type C pelvic ring lesions. Type B1-open book lesions occurred in 52 patients while 48 had lateral compression lesions of type B2 or B3. Only 13.1% of the patients showed no associated lesions. The frequency of associated neurological lesions in the B1 group with 21% was higher than in the B2/B3 group with 12%. The frequency of urologic lesions in group B1 was 9.6% but in B2/B3 patients it was only 2.1%. A total of 66% of B-lesion patients were stabilized with an external fixator. The frequency of secondary operative procedures was 23% in B1-patients and 6.25% in the B2/B3 group. Overall mortality was 10.4%: 5% in the B-group and 14.8% in the C-group. Outcome for patients with B2/B3 lesions was, with 92% excellent and good end results,much better than in patients with B1 lesions (74%). The inverse was true for radiologic results: 93% of the B1- but only 75% of the B2/B3-patients had an anatomical reduction. Open book lesions, lateral compression lesions and combined vertical stable lesions should be differentiated. We suggest that lateral compression lesions be characterized as B1 and open book lesions as B2 type injuries in the classification system of AO. PMID- 12883781 TI - [Employing a modified gamma nail for the treatment of proximal femoral fractures]. AB - A modified custom-designed long gamma nail (LGN10) and the long gamma nail (LGN11) were evaluated in a prospective trial for operative treatment of proximal femoral fractures. Between May 1998 and January 2000, 50 patients (mean age 79 years) were alternately treated with LGN10 or LGN11. The femoral antecurvature varied from 6 degrees to 10 degrees, with a maximum of 14 degrees, as evidenced by lateral radiograms. The need for intramedullary reaming in LGN10 cases was significantly less ( p=0.046). Other significant differences between the two study groups were not observed. One cranial cut out of the lag screw in the LGN10 group was observed. Other complications were not recorded during hospital treatment. Follow-up results are not yet complete. Using LGN10 seems to reduce intramedullary damage to the femur and make insertion of the nail easier. PMID- 12883782 TI - [Value of sonography to monitor the course of Achilles tendon rupture after treatment--comparison of conservative therapy, percutaneous tendon adaptation, and open suture]. AB - In this prospective study,73 patients (with an average age of 36 years) were examined by ultrasound to compare the results of open Achilles tendon surgery, percutaneous tendon adaptation, and conservative treatment. Once the accident reports had been collected, all of the patients presented themselves at the Sportklinik Stuttgart at intervals of 4, 8, 12, 26 weeks, and approx. 2.5 years after the accident. During the course of these examinations, the thickness and structure of the tendon was quantified via ultrasound and the quality by clinical examination. The results of the ultrasonic quantification of the healing process appeared to be similar in all patients. On the day of the accident the thickness of the tendon was 9.3 mm. Four weeks later a remarkable regeneration could be seen in all patients (12.0 mm), which reached a maximum (16.7 mm) after 26 weeks. By the end of the study, however, the result was a scarred and thickened tendon of 11.5 mm. The comparison of the ultrasound results and the clinical examination after 26 weeks and 2.5 years showed no correlation between the value of the echo structure and functionality of the tendon, and the comparison between the different therapy groups showed no significant difference in the healing process either. In conclusion, ultrasound examination is able to provide information concerning the stage of the healing process. The quality of the healing and healed Achilles tendon and the relationship to their function cannot be studied with ultrasound, and this is not influenced by the choice of therapy. PMID- 12883783 TI - [Long-term results after a modified Sauve-Kapandji procedure. Report on 105 post traumatic cases]. AB - A modified Sauve-Kapandji procedure was performed on 105 patients for a painfully limited range of motion and arthritis of the distal radioulnar joint following distal fracture of the radius ( n=81), the radius and ulna in the distal one third ( n=18) and of the forearm shaft ( n=6). After an average of 8 years all patients were followed up clinically (motion, strength, pain) and radiographically (union of the arthrodesis, carpal translation, radioulnar distance). Rotation of the forearm had been improved by 53%. The amount of strength lay by 70% in comparison to the contralateral side. In 97% of the patients pain could be reduced. In all cases the arthrodesis had fused completely. An ulnar drift of the carpus was observed in 5% of the patients, and 74% of the patients showed radiological signs of approximation of the proximal ulnar stump to the radius. This reduction of the radioulnar distance amounted to less than 3 mm in 65% of the patients and lay between 3 and 5 mm in 29% of the patients. In none of the cases was direct contact between the ulna and the radius encountered. PMID- 12883784 TI - [Non-operative treatment of thoracolumbar spinal fractures. Long-term clinical results over 16 years]. AB - Non-operative treatment of thoracolumbar fractures remains the standard treatment option for stable and slightly unstable spinal injuries at the level of the lower thoracic and lumbar spine. The purpose of the present retrospective study was to determine the outcome and long-term results of patients treated conservatively using either one of the two major non-operative treatment modalities: (1) functional, or (2) closed reduction and immobilisation by cast. The average follow-up time for the 43 patients reviewed was 16.3 years. They showed a radiologic increase in the average kyphosis angle of 5.2 degrees compared to the time of injury. No difference was noted between early functional therapy and treatment with closed reduction and immobilisation by cast. Patients with initial neurologic impairment showed a significant improvement ( P<0.05) in their neurologic status during the follow-up period. Results of validated psychometric questionnaires (SF-36, FSR, VAS-Spinescore) showed the characteristic pattern of a population with chronic back pain. In conclusion, a radiologic increase in the traumatic kyphotic deformity in patients with a non-operative treatment protocol has to be expected. Measurable negative physical and social long-term consequences can be anticipated after sustaining a type-A fracture of thoracolumbar vertebral bodies. Any correlation between radiologic and functional results has not been observed. PMID- 12883785 TI - [Diagnosis of tendon injuries to the hand]. PMID- 12883786 TI - [Hb 1.1 g/dl: successful treatment of a polytraumatized patient in hemorrhagic shock]. AB - We report on the successful treatment of a seriously traumatized patient (injury severity score=59) who presented with exsanguinating hemorrhage in the emergency room. Preclinical and emergency room management included "small volume" hypertonic resuscitation as well as massive transfusion, C-clamp stabilization of a pelvic ring instability, and attachment of external fixators to multiple bone fractures. During the intensive care period,we started enteral immunonutrition immediately after an abdominal compartment syndrome was removed by laparotomy and a jejunal catheter had been inserted. Multiple fractures were stabilized rapidly and the soft tissue wounds were covered by split skin grafts. Additionally, we provided sufficient analgesia with few side effects using an epidural catheter for continuous application of local anesthetics. In summary, an effective synthesis of up-to-date strategies for preclinical, emergency room, and intensive care management prevented multiple organ failure and achieved survival with good quality of life after a "fatal" multiple trauma. PMID- 12883787 TI - [FDG PET as an early nuclear medical diagnostic tool for aseptic wear-induced loosening of hip joint endoprostheses--a report of two cases]. AB - To date, 2-[F-18]fluoro-2-deoxy-D-glucose PET (FDG PET) is used as a tool in oncology as well as myocardiological and cerebral functional diagnostics in the clinical routine. False positive results of tumor search imply new possibilities for use in diagnostics of inflammation. The two case reports presented here on aseptic loosening of endoprostheses caused by the rub of polyethylene with histological and immunohistological refurbishing call attention to the possible diagnostic valency of FDG PET as an early warning system regarding aseptic artificial limb loosening induced by the rub of polyethylene. By quantification of glucose metabolism with the "standard uptake value" (SUV) as well as specific storage samples from around the artificial limb, it is the aim of our study group in further examinations to develop an algorithm which permits to distinguish between septic and aseptic loosening. PMID- 12883788 TI - [Treatment strategies for gastroesophageal reflux disease]. AB - Gastroesophageal reflux disease (GERD) is a very common disorder. Therapeutic options include lifestyle modifications, medical therapy, laparoscopic antireflux surgery, and three more recent options-injection therapy to the lower esophageal sphincter, endoscopic sewing procedures, and radio frequency ablation therapy. Medical therapy is effective in most patients but not always successful with advanced disease. Up to 70% of subjects do not have adequate nocturnal control of gastric acid secretion with 20 mg of omeprazole given twice per day. Patients who do not tolerate medical therapy, who respond inadequately, or who want to avoid life-long drug therapy are candidates for alternate treatments. Studies on endoscopic procedures such as polymethylmethacrylate (PMMA) injection, the Stretta procedure,and endoscopic suturing techniques all suffer from having small study groups for each procedure,unknown durability, short follow-up, and the absence of randomized, controlled procedures. Limitations on endoscopic techniques are esophageal motility disorders, severe esophagitis, and larger hiatal hernias. Laparoscopic antireflux surgery remains a well-established, durable alternative to long-term medical therapy. It has the benefits of convenience, safety, minimal complications, improved quality of life, and low cost. Alternative methods will have to earn their place against this gold standard. PMID- 12883789 TI - [Cocaine-body-packing. Infrequent indication for laparotomy]. AB - INTRODUCTION: Body-packers or "mules" are drug smugglers who swallow cocaine filled condoms in order to conceal them during air travel. Body pushers hide drug packages in the rectum or vagina. In a cooperative effort between the Frankfurt Airport Clinic and the GIZ-Nord (Goettingen University poison control center), we performed a retrospective study and developed an algorithm for the problem of "rupture of a cocaine-filled condom in a body-packer." METHODS: In a retrospective analysis, the data of all cocaine body-packers and body pushers who were identified at Frankfurt International Airport from 1985 to 2001 were evaluated.Temporal development, demographic data, and surgical aspects were of special interest. RESULTS: From 1985 to 2001 a total of 280 body pushers and 2880 body-packers were identified: 63 "mules" (2.2%) developed symptoms of severe cocaine intoxication following rupture of a condom. Emergency laparotomy was performed on 20 patients (i.e., 32% of all symptomatic body-packers) and the condoms were removed, while 43 body-packers (68%) died before surgical therapy could be initiated. All operated patients survived. CONCLUSION: Severe cocaine intoxication is life threatening. Patients die from complications caused by generalized vasoconstriction. If the reason for severe cocaine intoxication is the rupture of a cocaine-filled condom,the only possible therapy consists of immediate laparotomy for removal of the condoms. PMID- 12883790 TI - [Minimally invasive resections of gastric tumors]. AB - BACKGROUND: Submucosal and mucosal gastric tumors occur infrequently and have a highly variable clinical course. MATERIALS AND METHODS: Between 1996 and 2001, 44 patients with tumors of the stomach wall underwent a laparoscopic local resection in three different procedures and were analyzed prospectively. RESULTS: Laparoscopic resections were performed on 44 patients (21 female, 23 male). The mean age of the patients was 65.3 years (range: 23-78). We performed the laparoscopic intragastric resection (LIR) on 18, the full thickness resection (LVR) on 17, and the extramucosal enucleation (LER) on 9 patients. The laparoscopic resection was combined with endoscopy in 24 patients. The immunohistological examination of the specimens showed gastrointestinal stroma cell tumors (GIST) in 24 cases, leiomyomas in 10 cases, adenomas in 2 cases, neurinomas or benign neurofibromas in 4 cases, neuroendocrine tumors (NET) in 2 cases, and an ectopic pancreas islet in 1 patient. In all of these patients, the surgical margins in the pathological specimens were tumor-free without lymphatic or venous invasion. None of the patients with GIST or NET had distant metastases. Intraoperative complications occurred in three cases (6.8%). Two repeat laparoscopic procedures (4.6%) were necessary. No fatal outcome or relapse in the follow-up over 23.4 months (range: 1-74) was registered. CONCLUSIONS: Laparoscopic or laparoendoscopic resection of gastric tumors is practicable with relatively few of complications. The level of patient comfort is high. PMID- 12883791 TI - [Twenty-five years of experience in incisional hernia surgery. A comparative retrospective study of 432 incisional hernia repairs]. AB - INTRODUCTION: Incisional hernia surgery in Germany is changing from conventional techniques to mesh implantation. The relevance of different factors such as surgical technique, mesh material, and patient-related parameters concerning the outcome following mesh repair is still under debate. METHODS: In a comparative retrospective study of 432 incisional hernia operations on 348 patients we analyzed 11 autodermic hernioplasties, 241 Mayo procedures,and 180 mesh repairs over a 25-year time period. In addition to the quality of life following mesh implantation,the prognostic relevance of demographic, pre- and intraoperative parameters, surgical technique, mesh material, and the surgeon's experience were subjected to both univariate and multivariate analysis. RESULTS: With a mean follow-up of 9.7+/-8.8 years, the rate of major complications following mesh repair was 9% in contrast to 3% after the Mayo procedure (p=0.091). The sublay technique revealed less complications compared to the onlay procedure (p=0.016). The total recurrence rate following the overlapping Mayo repair was 37% in contrast to 15% after mesh implantation (p=0.001), with a significant superiority of the sublay technique over the inlay technique (p=0.043). The rate of recurrences and complications after autodermic hernioplasty was 72% and 36%,respectively. After mesh repair, 86% of the patients were better satisfied with the results after Marlex mesh compared to GoreTex (p=0.016). Mesh size was the only significant prognostic factor concerning quality of life following mesh implantation. The complication rate was determined significantly by the patients' risk factors, size of hernia, surgical technique, and the surgeon's experience, whereas the rate of recurrences was significantly influenced by the parameters obesity (BMI >25), size of hernia,and surgical experience. The recurrence rate decreased significantly with the surgeon's experience: a minimum of 16 mesh repairs led to a recurrence rate of less than 10%. CONCLUSIONS: Only the mesh repair revealed acceptable recurrence rates with high patient comfort. The sublay technique is superior to onlay concerning the complication rate, whereas the autodermic hernioplasty and inlay techniques are obsolete. The material of choice is polypropylene. The most important prognostic factor following mesh repair is the surgeon's experience. PMID- 12883792 TI - [Subtotal bilateral adrenalectomy preserving adrenocortical function]. AB - This study analyzed the frequency of recurrence and postoperative adrenocortical function in 16 patients who had been operated on by bilateral subtotal adrenalectomy since 1995. Bilateral pheochromocytoma was found in 13 patients, bilateral adrenal metastases in 2 patients, and bilateral micronodular adrenocortical hyperplasia with primary aldosteronism in 1 patient. An endoscopic approach was performed in four patients. The remaining 12 patients were operated on by an open approach. In ten patients, unilateral subtotal adrenalectomy with contralateral total adrenalectomy (synchronous or metachronous) was performed. Six patients underwent bilateral subtotal adrenalectomy. In all patients, a total of residual adrenal tissue of at least 1/3 of a normal adrenal gland was left in situ. 15 patients were successfully weaned from exogenous steroid substitution. During a mean follow-up period of 24 months, no recurrences were observed. Three patients died without local recurrence. The present study provided evidence for the safety and benefit of subtotal bilateral adrenalectomy, which could guarantee sufficient adrenocortical function in adrenal remnant volume of more than one third of one adrenal gland even after dividing the main adrenal vein. At our institution, particularly in patients with inherited pheochromocytoma, subtotal adrenalectomy has become a common surgical strategy. PMID- 12883793 TI - [Morbidity following simultaneous pancreas/kidney transplantation]. AB - BACKGROUND: Simultaneous pancreas-kidney transplantation (SPK) is still associated with the highest rate of morbidity among solid organ transplantations. Although improved long-term survival following SPK has been proven in IDDM patients, a further decrease in morbidity would be desirable. METHODS: A retrospective, single-center study was performed to investigate the morbidity following SPK and to compare the results to kidney transplantation alone (KTA). Parameters included the rates of relaparotomies, septic complications (urinary tract infection, wound infection, pneumonia), and graft function. RESULTS: Between September 2000 and August 2001, 99 patients underwent transplantation (34 SPK, 63 KTA, 2 pancreas transplants alone). Relaparotomies were performed in six SPK patients (17.6%), mostly due to complications related to the pancreatic graft (n=5). Three reoperations (4.8%) were necessary in KTA patients (p=0.085). Septic complications occurred more often in SPK than in KTA patients (55.9% vs 30.2%, p<0.05). This difference resulted from the high rate of wound infections in SPK patients (35.3%). No intra-abdominal infection or sepsis occurred in any patient. There was one hospital death in SPK and KTA patients, respectively. The rejection rate was similar in SPK (17.6%) and KTA (12.7%, p=0.72). At discharge 91.2% of SPK patients were insulin free and 97.1% free of dialysis. At discharge 96.8% of KTA patients were free of dialysis. CONCLUSION: SPK is still associated with a higher morbidity (relaparotomies, septic complications) than KTA, although life threatening complications were rare. There was no increased mortality following SPK. PMID- 12883794 TI - [Clinical usefulness and perspectives of sentinel node biopsy in the management of breast cancer]. AB - INTRODUCTION: In view of its predictive potential, axillary nodal status plays a particularly important role in breast cancer. The concept of sentinel node biopsy (SNB) revealed an accurate method with low postoperative morbidity for staging the axilla in patients with lymph node-negative breast cancer. The aim of this study was to show that SNB alone must have a place in routine clinical work and is reliable after preoperative chemotherapy (PC) and also in patients with multicentric tumors (MC). PATIENTS AND METHOD: Between April 1997 and March 2002, a total of 300 SNBs were performed in patients with uni- or bilateral breast cancer. Of them, 45 who had completed PC and 13 with multicentric lesions underwent SNB followed by axillary dissection. The sentinel nodes (SN) were labelled with a blue dye and radioactive colloids. Median follow-up was 33.6 months (range 8.2-67.0) (StAw 1.4). RESULTS: The detection rate in the learning phase was 81.8% and later 93.5%, independently of the size of the primary tumor. Overall accuracy and negative predictive value of the first 55 patients, after PC and in those with MC came to 97.8% and 96.7%, 97.6%, 95.8% and 100%, respectively. Through focused pathologic examination, staging was improved in 11.3% of patients. A year after the procedure, one patient developed recurrence. Morbidity after SNB alone was significantly lower than after axillary dissection. CONCLUSION: Our experience shows SNB to be reliable and accurate for axillary staging in breast cancer patients. Morbidity after SNB alone was low. Conclusive data on the local recurrence rate are not yet available. Under certain conditions, SNB appears to have future clinical potential, even in patients with PC and MC, which must be confirmed in further multicentric studies. In the meantime, this method has attained definite value in the surgical therapy of breast cancer patients. PMID- 12883795 TI - [Sentinel lymph node dissection in patients with malignant melanoma. Diagnostic and therapeutic standards]. AB - INTRODUCTION: In patients with cutaneous malignant melanoma, the sentinel lymph node (SLN) reflects the histopathological features of the lymphatic basin with high accuracy. MATERIAL AND METHODS: Three hundred eighty-one melanoma patients at the Hornheide clinic with an overall follow-up of 36 months (November 1998 to October 2001) underwent sentinel lymph node dissection (SLND). RESULTS: The SLNs were successfully found in 93% of truncal melanoma ( n=136), 97% of melanoma of the extremities ( n=184), and 86% of melanoma of the head and neck region ( n=61). Of truncal midline melanomas, 84% ( n=43) showed two or more regional basins, in contrast to 18% of nonmidline melanoma ( n=93). Histopathological analysis revealed occult nodal disease in 25% of all patients. Completion lymphadenectomy revealed residual nodal disease in 8% of all patients with low risk melanoma with a tumor thickness of 0-1.5 mm (two of 26 patients with positive SLN) and in 11% of all patients with high risk melanoma with tumor thickness above 1.5 mm (eight of 70 patients with positive SLN). Tumor relapse was noted in 5% of negative SLN patients and 14% of positive SLN patients. The results of the method were false negative in 2% with a sensitivity of 98%. CONCLUSION: Sentinel lymph node dissection is a reliable and accurate method of staging regional lymph nodes for all primary tumor sites. It can localize occult metastases in unexpected lymphatic basins and provides critical indications for completion lymphadenectomy. It represents an essential method of establishing stratification criteria for future adjuvant trials. Further long-term follow-up is needed to investigate its prognostic relevance to recurrence and overall survival. PMID- 12883796 TI - [Coverage of pressure sores with free flaps]. AB - INTRODUCTION: The coverage of recurrent pressure sores with unstable scar in the surrounding tissue is still an unsolved problem in the literature. Local and regional transfer of tissue often does not meet the requirements of the tissue deficit. Especially in recurrent pressure sores, the adjacent skin has already been consumed due to multiple surgeries. As a good alternative, the microsurgical transfer of flaps offers viable tissue to cover even large pressure sores. METHODS: We performed a total of six free flaps in five patients who suffered from intractable pressure sores in the hip region. The age of the patients was between 41 and 63 years. The defect size varied between 6 x 6 cm and 25 x 30 cm. Two combined myocutaneous scapula-latissimus dorsi, two myocutaneous latissimus dorsi, one anteromedial thigh, and one rectus femoris flap were used to cover the defects. RESULTS: The average follow-up time was 29 months. Flaps provided stable coverage in four of five patients at 12-month follow-up. There was one subtotal flap necrosis that was subsequently treated with split-thickness skin grafting. CONCLUSION: In this series of five patients with six free flaps, we were able to show that the microsurgical transfer of tissue is a valuable option in the treatment of difficult pressure sores. Even in older and debilitated patients, this method is a good alternative to conventional local flaps. PMID- 12883797 TI - [Psoas abscesses. Genesis, diagnosis, and therapy]. AB - BACKGROUND: A psoas abscess is a rarely encountered entity with various etiologies and nonspecific clinical presentation, frequently resulting in delayed diagnosis, increased morbidity, and prolonged or recurrent hospitalization. PATIENTS AND METHODS: Between January 1996 and January 2002 we treated ten patients (approximately 54.8, 5 males,5 females). These cases were analyzed retrospectively relative to a review of the literature. RESULTS: CT scanning was decisive in the final diagnosis of psoas abscess. Primary psoas abscess occurred in four cases and six patients had secondary abscesses. In all except one case, the psoas abscess was located on the right side. The causes of primary abscesses were retroperitoneal perforated appendicitis, paravertebral injections for lumboischialgia, Pott's disease, and repeated intravenous drug application in the groin. Five patients underwent retroperitoneal open drainage and four patients CT guided drainage. One patient with retroperitoneal perforated appendicitis was treated by laparotomy. Staphylococcus aureus, Bacteroides fragilis, and Escherichia coli were the most common infective agents. There was no postoperative mortality and no cases of abscess recurred. CONCLUSIONS: CT scan is a diagnostic "gold standard" for psoas abscess. CT-guided drainage is the method of first choice, but is not possible in all cases. Open retroperitoneal drainage is a standard method of treatment. Postoperative antibiotic therapy is obligatory and should be adapted individually. PMID- 12883798 TI - [Sugar tumor of the lung--case report and review of the literature]. AB - Clear cell tumors of the lung are commonly primary, clear-cell, bronchial carcinomas or metastasis of a renal cell carcinoma. Compared to this, pulmonary sugar tumor is a rare entity. A large intracellular content of glycogen and immunohistochemical procedures lead to diagnosis. The demonstration of the premelanosomal protein HMB-45 is considered as proof, but this is not airtight. We present a case of metachronic, benign, and HMB-45-negative sugar tumor of the lung after hypernephroma and give a review of the literature. PMID- 12883799 TI - [Quality management in surgery--an overview of methods and possibilities]. AB - Numerous methods of assessing quality in health care have been presented over the last years. Since the German health care reform of 2000 introduced a legal obligation to quality assurance, hospitals have to meet that condition. However, no specific system of quality assurance was implicated. Therefore, many potential users search for systems suiting their own needs. However, German hospitals can benefit from international experience in quality management. Accreditation, visitation, excellence schemes, and certification seem to be the dominant methods. Every method is worthy of hospital evaluation. Certifications illustrate the quality system view without regard to profession, while excellence schemes evaluate the development of quality management in hospitals. Accreditation portrays a specific hospital as a whole, while visitations follow the professional perspective of quality assurance. Depending on the perspective and aims, there are advantages and disadvantages in each method. Finally, a combination of two may also be helpful. Indeed, no "role model" exists yet for assessing quality in health care. These methods of quality management are applicable especially in surgery, because a variety of standard outcome parameters such as length of stay, duration of procedure, and complication and relapse rates are already available. PMID- 12883800 TI - [Living organ donor transplantation--the German experience in comparison to others]. AB - Living liver donation is a prominent and innovative method in the therapy of terminal liver disease in children and adults. Live donation of livers was first used in 1989 for children. After only 10 years, the concept was also established for application in adults. Due to the imminent lack of donor organs and the constantly increasing need of liver transplantation, this method will continue to grow in importance. PMID- 12883801 TI - [Living kidney transplantation. A comparison of Scandinavian countries and Germany]. AB - The discussion of compensating for shortages of cadaveric donation with increased living donation often reveals differences between the Scandinavian countries and Germany. Possible adoption of Scandinavian structures to improve the rate of living donations in Germany warrants analysis of the actual differences between these two regions. Close examination reveals that significantly higher rates of living donation are achieved only in Sweden and Norway. In Norway, a frequently postulated negative effect on cadaveric donation due to very high rates of living donation could not be confirmed. In contrast to Germany and as a consequence of Norwegian geography, kidney transplantation has been regarded in Norway as the first-line therapy for endstage renal disease for more than 35 years. Living donation has since been actively pursued and is traditionally the transplantation of first choice. In Germany, living donation is still regarded as the second choice after cadaveric donation, due to legal regulations. Significant improvements in living donation frequencies could be achieved there by adopting the active Norwegian approach to living donor identification. PMID- 12883802 TI - [Basic ethical aspects of living organ donation]. AB - A characteristic feature of transplanting organs from living donors is that not only patients in need for treatment but also healthy individuals are submitted to medical interventions. Ethical considerations in this field have to deal with the question of property attributes of the human body and conflicts with traditional medical principles. Altruistic organ donation, appreciated by Christianity as a sign of charity, is indeed contradictory to the classic maxim of medical ethics "primum nihil nocere, " meaning "first of all, do not harm." The autonomous choice of a potential donor has to be balanced thoroughly against his personal physical and psychological risks. Apart from organ donation with altruistic motives, commercial incentives or payment for organ donation, which are increasingly under discussion in many nations, need profound ethical reflection. Organ selling does not lead to long-term economic benefit for individual donors in developing countries and is associated with a decline in health. A market system of organ sales would foster exploitation of the poor, and it is substantially doubtful whether autonomy and self determination are valid under circumstances of poverty and coercion. Commodification of the human body risks viewing persons as marketable objects. The human body,however, is an integral element of an individual's personality and not a resource to be removed. It is therefore fundamental that the social good of altruism is preserved as the major principle in organ donation. PMID- 12883803 TI - [Innovations in liver surgery through transplantation from living donors]. AB - Living-donor liver transplantation was introduced into clinical practice in the early 1990s. At first the results were unsatisfactory, but today's results after living donation are as good as those obtained after conventional liver transplantation with full-sized organs. With minimally invasive diagnostic methods, it is now possible to determine the quality of potential donor livers and exclude focal lesions and anatomical variants which influence the strategy of organ retrieval procedures. Donor liver resection is done without hilar occlusion after determining the anatomical variants of the bile system (especially for right lobes) and localizing of the course of the middle hepatic vein. Microsurgical techniques are used for reconstruction of the biliary system and hepatic vessels. Living-donor liver transplantation allows us to investigate the complex changes after liver resection. The surgical techniques and pathophysiological postoperative changes can be adapted unconventionally to complex oncological liver resections. Therapeutic optimization and better risk management are becoming possible for liver resections. PMID- 12883804 TI - [Reconstruction of the inferior vena cava for extended resection of malignant tumors]. AB - OBJECTIVES: Malignant tumor invasion in the inferior vena cava (IVC) has for a long time been the limiting factor in the resection of retroperitoneal tumors. The clinical outcome in these patients depends on vascular surgical techniques, the central role of which is played by IVC reconstruction. METHODS: Within the last 7 years, 9,085 vascular reconstructive procedures were performed in our department. Six patients suffered from retroperitoneal invasion of tumor into the IVC. After tumor resection, the involved IVC segments were replaced by polytetrafluorethylene (PTFE) grafts to restore IVC continuity. In three patients, an adjunctive arteriovenous (AV) fistula was constructed. RESULTS: The graft patency after a mean follow-up of 30.2 months (range 1 to 79) was 83.3%. The only graft occlusion occurred in a patient without AV fistula. There were no perioperative deaths and no major complications demanding reoperation. CONCLUSION: In patients with tumor involvement of the IVC, clinical outcome depends on vascular surgical coprocedure. After resection of the IVC, a PTFE graft should be interposed in combination with an AV fistula. Anticoagulation and CT scan are recommended after 3 months before ligation of the AV fistula. PMID- 12883805 TI - [Refixation of osteochondral fragments using absorbable implants. First results of a retrospective study]. AB - An osteochondral lesion in a weight bearing joint causes therapeutic problems. Surgical therapy focuses on the restoration of the articular surface, unlimited motion of the joint and prevention of cartilaginous degeneration. A causal therapy to prevent posttraumatic osteoarthritis is the fixation of osteochondral fragments. Various absorbable implants for the purpose of refixation are available, but only a few have proved to be biocompatible. This retrospective study presents the preliminary results after refixation of osteochondral fragments of the ankle and knee joints. We used self-reinforced absorbable pins, nails and screws made of polylactide acid. The results for 15 patients with 17 fractures of 16 joints were evaluated. All patients were scored by the Tegner activity-level, the McDermott-score and the DGKKT-score, and 13 of them were monitored by postoperative magnetic resonance imaging (MRI). The follow-up extended for an average of 14.3 months (range: 4-43 months). The clinical results for all patients were good (McDermott-score: O 89 points, DGKKT-score: O 78.8 points, Tegner: pre-op vs post-op=O 4.6 vs O 5.1) and the surgical therapy proved to be successful. The MRI showed the complete incorporation and vitality of the fragment as well as a congruence of the joint surface. The clinical course and the MRI gave little reference to an inflammatory reaction due to the implants that complicated the course of two patients. These results indicate that absorbable implants made of polylactide acid are biocompatible and effective in the fixation of osteochondral fragments. PMID- 12883806 TI - [Anterior semifundoplication in the treatment of gastroesophageal reflux disease: long-term results following conventional surgery]. AB - INTRODUCTION: According to randomized studies, semifundoplication in the treatment of gastroesophageal reflux disease (GERD) is related to lower postoperative dysphagia rates than with fundoplication in comparable reflux controls. However there is a lack of long-term results. The object of this study was therefore to determine the influence of partial anterior fundoplication on the long-term clinical outcome (>1 year) in patients with GERD. METHOD: From December 1986 until May 2000, 100 patients suffering from GERD were operated on. Seven of them were not evaluated, four because of revisional surgery, two because of preceding multiple abdominal operations (MEN, colitis ulcerosa), and one because of Nissen fundoplication. In all, the perioperative results of 93 patients (51 men and 42 women aged 21 to 86 years) were evaluated. Fourteen patients died during the follow-up period and two were lost because of changes in address. Thus, 77 patients with a median follow-up of 88 months (range 15-94) were interviewed with a list of standardized questions concerning reflux control and dysphagia. RESULTS: The median operation time was 110 m (range 55-270). In one patient, an esophageal mucosal tear was detected intraoperatively and promptly repaired. Postoperatively, 71.4% (55/77) had no reflux complaints, 85.7% (66/77) had dysphagia, 66.2% took no further medication, and 31.2% (24/77) continued taking medication. The median interval free of symptoms was 25.5 months, and in 40.9% of the patients, symptoms recurred within the first year after operation. Five patients were in need of reoperation. A total of 77.9% of the patients were satisfied with the results of the operation. CONCLUSIONS: With regard to reflux control, long-term results of anterior semifundoplication are comparable to those of fundoplication. However, considering postoperative dysphagia, the technically easier anterior semifundoplication is less eventful and therefore a good alternative which in the long run shows good results after laparoscopic intervention. PMID- 12883807 TI - [Arterial back table pressure perfusion prevents ischemic biliary lesions after orthotopic liver transplantation]. AB - INTRODUCTION: Ischemic biliary lesions are a threatening complication following orthotopic liver transplantation. Their exact pathophysiological mechanism is unknown so far, but insufficient perfusion of biliary arterial vessels might be responsible for the development of these lesions. This might be changed by improved perfusion techniques. We performed a controlled study of cases since February 2000. MATERIALS AND METHODS: We used arterial back table pressure perfusion to achieve reliable perfusion of the capillary system of the biliary tract, which may be impaired by the high viscosity of University of Wisconsin solution. In this study, 190 orthotopic liver transplantations performed between September 1997 and July 2002 were investigated with regard to ischemic biliary lesions. RESULTS: One hundred thirty-one grafts were preserved by in situ standard perfusion including portal perfusion,whereas additional arterial back table pressure perfusion was performed in 59 cases. Donor-related factors, recipient age, indications for transplantation, transplantation techniques, and ischemia times were comparable between groups. Twenty-one (16%) of the patients in the standard perfusion group and only one of the those receiving arterial back table pressure perfusion developed ischemic biliary lesions. This difference was highly significant (P=0.004). Maximal aspartate aminotransferase and alanine aminotransferase levels in the first 3 days were significantly lower in the arterial back table pressure perfusion group (P>0.05). CONCLUSION: Arterial back table pressure perfusion is an easy and reliable method for preventing ischemic biliary lesions in orthotopic liver transplantation. It should, therefore, be the standard technique in liver procurement. PMID- 12883808 TI - [Collateral circulation of the left colon: historic considerations and actual clinical significance]. AB - INTRODUCTION: The arterial communication between the superior and inferior mesenteric arteries is important in surgery of the colon and aorta. METHODS: The anastomosis between the median colic artery (ACM) and the superior colic artery (AMS) was studied on 52 cadavers (32 female, 20 male). The length of the artery was measured using a flexible scale, and its diameter was determined at three different locations (origin, middle, end). RESULTS: The median lengths of the arteries were: 6.8 cm (range 3.9-9.7) for ACM, 12.2 cm (range 7-17.4) for ACS, and 23 cm (range 13.6-34.2) for margin artery. The median diameters were: 3.4 mm (range 2.7-4.1) for ACM, 3.1 mm (range 2.1-4.1) for ACS, and 2.7 mm (range 2-3.4) for margin artery. In 9.6% of the cadavers ( n=5), additional anastomoses between the AMS and inferior mesenteric arteries were proven. In 90.4% ( n=47), only margin arteries were found. SUMMARY: The low incidence of an additional communication (Riolan's arch) and possible additional impairment of the blood supply should be taken into account during operation. PMID- 12883809 TI - [Tracheal rupture from blunt chest injury in a child]. AB - Tracheobronchial lesions after blunt chest injury are rare. Diagnosis and therapy can prove to be challenging for the attending physician. Nonrecognition of this injury may entail severe complications. Herein we report the case of a 9-year-old girl with rupture of the trachea due to a fall against the handlebar of her kickboard. The clinical, radiological, and features of the therapy performed are discussed. PMID- 12883810 TI - [Heterotopic gastric pancreas: a rare cause of abdominal complaints]. AB - Heterotopic pancreas is an uncommon cause of gastrointestinal complaints such as epigastric pain, nausea, vomiting, and upper gastrointestinal bleeding. Despite the development of modern diagnostic procedures, it is still difficult to differentiate heterotopic pancreatic tissue from other benign or malignant gastric tumors. Local excision of the gastric wall is regarded as the diagnostic and therapeutic procedure of choice. We present two cases and an overview of the literature. PMID- 12883811 TI - [Anti-inflammatory therapy in sepsis]. PMID- 12883812 TI - [Surgical treatment of hyperparathyroidism]. PMID- 12883813 TI - The endoscopic stapler diverticulotomy for Zenker's diverticulum. AB - This paper describes the surgical procedure of the endoscopic stapler treatment of Zenker's diverticulum and analyzes the results of 24 consecutive operated patients. In three patients the endoscopic exposure of the diverticulum was not possible. Twenty-one patients underwent endoscopic stapler treatment without any peri- or postoperative complications. The follow-up period was 4 to 29 months (average 18 months). The average total time for surgery was 25 min. Postoperatively, a nasogastric feeding tube was not necessary: all patients resumed oral intake 12 h after surgery. Discharge from the hospital followed the 2nd postoperative day. All patients had complete or nearly complete resolution of symptoms at the 4-month follow-up. Recurrent complaints were an indication for repeat of the contrast barium esophagram. Two patients revealed a residual diverticulum 7 and 11 months after treatment. In comparison with results and complication rates in the literature of the external, transcutaneous techniques and endoscopic diverticulotomy procedures, the endoscopic stapler treatment of Zenker's diverticulum is a safe, (cost-)effective and minimally invasive method and to be considered as the initial treatment of choice. PMID- 12883814 TI - Can low frequency sound stimulation during posturography help diagnosing possible perilymphatic fistula in patients with sensorineural hearing loss and/or vertigo? AB - Perilymphatic fistula (PLF) is often difficult to diagnose because of the similar symptomatology, such as vertigo, tinnitus and hearing loss, which is found in several inner ear diseases. We attempted to correlate a positive result of low frequency sound (LFS) stimulation tests in posturography with the presence or absence of a PLF confirmed by transtympanic endoscopy in 209 patients with various inner ear diseases (Meniere's disease ( n=128), vestibulopathy ( n=41), cochleopathy ( n=28) and sudden deafness ( n=12). LFS provoked unsteadiness in posturography without PLF in 24 patients with Meniere's disease, in 5 patients with vestibulopathy, in 3 patients with cochleopathy and in 2 patients with sudden deafness. In one patient, tympanoscopy revealed fistula in the round window membrane that was covered with a fibrinous layer. In four cases there was abnormal light reflex in the round window but without PLF. In eight cases, Hennebert's sign was present with nystagmus, without PLF. We conclude that pathological responses to the LFS test in posturography can also be encountered in other inner ear diseases without PLF. PMID- 12883815 TI - Congenital vomeral bone defect in two thalassemia trait cases. AB - The nasal septum is composed of a perpendicular plate of ethmoid bone, quadrilateral septal cartilage, membraneous septum, crest of palatine and maxillary bone and vomer. Defects of the nasal septum may be due to a variety of causes such as trauma, infection, inhalant irritants and neoplasia. To our knowledge, up until now, congenital defect of the vomer has been presented in six cases without any nasal symptoms. We present two cases of congenital defect of vomer with thalassemia trait, the patients complaining of nasal obstruction. PMID- 12883816 TI - Voice quality change in future professional voice users after 9 months of voice training. AB - Sixty-eight students of a school for audiovisual communication participated in this study. A part of them, 49 students, received voice training for 9 months (the trained group); 19 subjects received no specific voice training (the untrained group). A multidimensional test battery containing the GRBAS scale, videolaryngostroboscopy, Maximum Phonation Time (MPT), jitter, lowest intensity (IL), highest frequency (FoH), Dysphonia Severity Index (DSI) and Voice Handicap Index (VHI) was applied before and after training to evaluate training outcome. The voice training is made up of technical workshops in small groups (five to eight subjects) and vocal coaching in the ateliers. In the technical workshops, basic skills are trained (posture, breathing technique, articulation and diction), and in the ateliers, the speech and language pathologist assists the subjects in the practice of their voice work. This study revealed a significant amelioration over time for the objective measurements [Dysphonia Severity Index: from 2.3 to 4.5 ( P<0.001)] and the self-evaluation [Voice Handicap Index, from 23 to 18.4 ( P=0.016)] for the trained group only. This outcome favors the systematic introduction of voice training during the schooling of professional voice users. PMID- 12883818 TI - Pleomorphic adenoma of the parotid gland: a 13-year experience of consequent management by lateral or total parotidectomy. AB - Optimal surgical management of parotid pleomorphic adenoma is important because of a considerable risk of tumour recurrence and complications. Our series of primary surgery cases with standardised parotidectomy technique in a university hospital were evaluated. Medical records of 295 patients treated from 1987 to 1999 were reviewed, and 171 patients answered a questionnaire about long-term sequelae. Sixty-two patients reporting major complications were re-examined in the hospital. At a mean follow-up of 8 years, one patient (0.6%) re-appeared with a recurrence. Twenty-four percent left the hospital with a partial and 3% with a complete paresis. Only 5% developed a permanent partial paresis, whereas a permanent total paresis was not seen. Half of the patients indicated gustatory sweating, but treatment was required in only 6%. Other major permanent complications were not seen. Treatment of pleomorphic adenoma by standardised parotidectomy in a university hospital demonstrates favourable results with low perioperative and long-term morbidity, as well as minimal risk of tumour relapse. PMID- 12883819 TI - Primary mucosal melanoma of the eustachian tube. AB - Melanomas of the upper airway tract constitute around 1% of the total melanomas of the head and neck. They are usually more advanced in depth of invasion or size at the time of diagnosis than melanomas of the skin. The case of a 74-year-old Caucasian female with primary malignant melanoma of the left eustachian tube and bilateral neck metastasis is presented. The procedure from the diagnosis of neck metastasis to the location of the primary melanoma is described. The management of the malignant melanoma neck metastasis should also involve endoscopic examination of the upper aerodigestive tract, since the primary lesion can occur there. PMID- 12883820 TI - Hearing preservation in perilymphatic fistula due to a congenital fistula in an adult. AB - Congenital fistula in the stapedial footplate usually can be diagnosed by the recurrence of meningitis caused by spontaneous cerebrospinal fluid (CSF) in infants or young children. We report the case of a 65-year-old woman, who was initially diagnosed as having an acquired perilymphatic fistula caused by aural barotrauma and demonstrated episodic vertigo and fluctuant sensorineural hearing loss in the right ear after air travel. Surgical exploration showed a congenital circular defect in the peripheral part of the stapedial footplate with leakage of CSF. The fistula was closed by inserting a tiny piece of fascia attached to both the tympanic and perilymphatic side of the stapedial footplate utilizing the back pressure of perilymphatic fluid and fibrin glue; hearing was preserved. PMID- 12883821 TI - CAP amplitude after impulse noise exposure in guinea pigs. AB - In this study, 21 guinea pigs were submitted to a single high energy impulse noise (gun shot with blank projectiles). The auditory function was evaluated over a 7-day recovery period by recording the compound action potential (CAP) from the round window. The threshold shift and input/output function (CAP amplitude and delay function of the stimulus intensity) were studied at different frequencies. CAP amplitude fell after the noise trauma, especially at the lower sound level, resulting in a threshold shift. Latency was significantly increased. During recovery, whereas latency returned to its initial value, CAP amplitude gradually increased and, in half the animals, exceeded the control value for the higher levels of stimulus. This could have been because of progressive disinhibition or recruitment, and may correspond clinically to hyperacusis. These results are discussed referring to those obtained by other authors using other methods. PMID- 12883822 TI - Indirect autofluorescence laryngoscopy in the diagnosis of laryngeal cancer and its precursor lesions. AB - Direct autofluorescence endoscopy of the larynx has proven to facilitate the detection and delineation of precancerous lesions, carcinoma in situ and cancer. The aim of the present study was to evaluate the diagnostic potential and limitations of this imaging technique applied during indirect laryngoscopy. In a prospective study, 116 patients with suspected precancerous or cancerous lesions were investigated preoperatively by indirect autofluorescence laryngoscopy. Autofluorescence was induced by filtered blue light (375-440 nm) of a xenon short arc lamp and processed by a CCD camera system (D-light-AF System, Storz, Tuttlingen, Germany). Autofluorescence images were immediately assessed for diagnosis, compared to the direct autofluorescence picture obtained during microlaryngoscopy and compared to pathohistological findings. Comparable to direct autofluorescence images, normal laryngeal mucosa showed a typical green fluorescence signal. Moderate and high epithelial dysplasia, carcinoma in situ and cancer displayed a diminished green fluorescence. False negative results were related to extreme hyperkeratosis. False positive cases showed mild dysplasia with inflammatory reactions or scarring. In 103 cases (89%), we found concordant results. According to our results, the presented technique seems to be a promising diagnostic tool for the early detection of laryngeal cancer and its precursor lesions during indirect laryngoscopy. Scarring, marked hyperkeratosis and inflammation can limit the predictive value of the method. PMID- 12883823 TI - Gonadotropin (LH, FSH) levels in serum and cyst fluid in epithelial tumors of the ovary. AB - OBJECTIVES: The aim of this work was to determine gonadotropin (LH, FSH) levels in serum and cyst fluid in various type of ovarian epithelial neoplasms (benign, borderline, malignant) and to compare them with levels in benign cysts. Additionally we decided to estimate if there were some significant correlations between serum and ovarian cyst fluid in gonadotropin levels in all investigated groups. MATERIALS AND METHODS: The study group included 74 patients before (n=36) and after (n=38) menopause, divided into four groups depending on the histopathologic diagnosis. Serum and cyst fluid levels of LH and FSH were determined in all patients. RESULTS: We found statistically significant differences concerning LH and FSH levels in serum and cyst fluid between malignant and borderline tumors, between malignant tumors and benign cystadenomas and between malignant tumors and non-neoplastic cysts. We also found statistically significant correlations between serum and cyst fluid as to FSH and LH levels in patients with borderline tumors. There were no statistically significant correlations between serum and cyst fluid as to FSH and LH levels in patients with malignant epithelial tumors (group 1) and in patients with benign cystadenomas (group 3). CONCLUSIONS: High FSH and LH levels in cyst fluid of malignant epithelial tumors of the ovary are in line with the gonadotropin theory of tumor growth. Simultaneous determination of various types inhibin levels appears to be an interesting topic for our future research. PMID- 12883824 TI - Immunohistochemical expression of inhibin-alpha in human endometrium and the in vitro secretion of inhibin, estradiol and cortisol in cultured human endometrial glandular cells. AB - BACKGROUND: Inhibins are multipotent dimeric glycoproteins, composed of an alpha subunit and one of two possible beta-subunits (betaA or betaB). Aims of this study were (a): the immunohistochemical characterisation of normal human endometrium for the inhibin-alpha subunit; (b) the assessment of the secretion and metabolism of inhibin, E2 and cortisol; (c) the evaluation of any relationship between these three substances in cell culture medium of isolated and cultivated normal human endometrial glandular cells. MATERIALS AND METHODS: Samples of human endometrium were obtained from 34 premenopausal patients. Nineteen endometrial specimen (proliferative [PP] n=8; early secretory [ES] n=7; late secretory phase [LS] n=4) were brought into cell culture. Fifteen endometrial specimen (PP n=5; ES n=5; LS n=5) were paraffin-fixed and used for the immunohistochemical analysis for inhibin-alpha. Stromal and epithelial cells were separated by collagenase digestions, filtrations, sedimentations and Ficoll gradient centrifugation. E2 and cortisol were measured with radioimmunoassay (RIA) and inhibin with enzyme-immunoassay (EIA). Statistical analysis was performed with the non-parametric Mann-Whitney rank-sum test and linear regression analysis. RESULTS: Inhibin-alpha showed a weak (positive) expression during proliferative phase, which increased significantly as the menstrual cycle continued. In secretory glands the mean inhibin concentration was higher than that from proliferative samples. A significant correlation was observed between inhibin and E2 (p<0.001) as well as cortisol and inhibin (p<0.0001) in glands from proliferative phase. Between inhibin and E2 (p<0.05) as well as inhibin and cortisol (p<0.002) a significant correlation in early secretory glands was also noted. In late secretory phase inhibin and E2 (r2=0.78650; p<0.0001), inhibin and cortisol (r2=0.58326; p<0.001) and E2 and cortisol (r2=0.52880; p<0.001) showed a significant correlation. DISCUSSION: In conclusion, we found a cyclical expression of inhibin-alpha subunit in the endometrium demonstrated by immunohistochemical means. A higher in vitro secretion of inhibin from secretory glands was also observed. In addition, a significant correlation between inhibin with E2 and cortisol in PP and ES glands and a significant correlation between inhibin, E2 and cortisol in LS glands could also be demonstrated. We conclude that inhibin can be associated with E2 and cortisol metabolism, playing an important role in paracrine/autocrine mechanisms in the endometrium and possibly exerting its function through cortisol and E2. The cortisol concentration also correlates with E2, suggesting a link between these steroids in the endometrial function. The correlation of inhibin, E2 and cortisol suggest complex autocrine/ paracrine mechanisms in human endometrial glands, modulated and controlled by all these three substances. PMID- 12883825 TI - Maternal and neonatal morbidity of emergency caesarean sections with a decision to-delivery interval under 30 minutes: evidence from 10 years. AB - PURPOSE: The purpose was to investigate the decision-to-delivery interval for emergency caesarean section and to compare the perioperative maternal and neonatal morbidity to that of intrapartum non-emergent caesarean section. MATERIALS AND METHODS: A cohort study was designed evaluating 109 women undergoing 'crash' emergency caesarean section and 109 controls from 1988 to 1997 in a University hospital. All emergency caesarean sections were performed in the delivery room. Controls were matched by gestational age. Retrospective collection of data relating to indication, maternal and neonatal outcome of both groups was conducted. Time intervals between decision to deliver and actual delivery were collected for all emergency deliveries. RESULTS: The emergency caesarean section cohort represented a high risk group with a mean gestational age of 34.8 weeks and a high number of patients with preterm labour (29.3%). All 109 emergency caesarean sections were achieved within a decision-to-delivery time of 30 min. Severe maternal haemorrhage was significantly higher mainly because of underlying obstetrical complications such as placenta praevia, placental abruption, and others resulting in a blood transfusion rate of 10.1%. Procedure related and infectious maternal morbidity, however, was not significantly increased. All women undergoing the emergency procedure received perioperative antibioprophylaxis. Mean umbilical arterial blood pH and Apgar scores were significantly lower in infants delivered by emergency caesarean section but there was no difference in the admission to the NICU. CONCLUSIONS: Emergency caesarean section performed in the delivery room may result in a decision-to-delivery time of less than 30 min without detrimental perioperative effects on mother or infant. PMID- 12883826 TI - Inhibitory effect of catechin against the superantigen staphylococcal enterotoxin B (SEB). AB - Staphylococcal superantigens (SsAgs) have gained attention as one of the factors aggravating atopic dermatitis (AD) and several potential mechanisms of AD aggravation by SsAgs have been reported. Tea catechin has been found to have many unique antimicrobiological activities such as antibacterial, antiviral, antifungal and antitoxic effects. In the present study, we investigated the inhibitory effect of the green tea catechin extract, Polyphenon, and (-) epigallocatechin gallate (EGCg) on staphylococcal enterotoxin B (SEB) and its mechanisms of action, and we also discuss the possibility of therapeutic benefits for AD patients of tea catechin. Polyphenon inhibited the lethal toxicity of SEB and the SEB-induced production of TNF-alpha, IFN-gamma and IL-4 following its intraperitoneal administration to BALB/c mice. Although Polyphenon is composed of several isomers among which EGCg is approximately 50% of the total, we considered that most of the inhibitory effect of Polyphenon in mice could be attributed to EGCg. EGCg was immediately bound to SEB molecules and neutralized SEB in a dose- and incubation time-dependent manner without molecular weight alteration of the SEB molecule. Furthermore, EGCg inhibited SEB-induced TNF-alpha and IFN- gamma production and IL-2, IFN-gamma, IL-10 and IL-12 p40 mRNA expression in human PBMCs from normal donors in a dose-dependent manner. Inhibition of SsAg-induced T cell activation by catechin was observed in both in vivo and in vitro studies, suggesting that catechin may be useful in the treatment of AD. PMID- 12883827 TI - ECP-treated lymphocytes of chronic graft-versus-host disease patients undergo apoptosis which involves both the Fas/FasL system and the Bcl-2 protein family. AB - Chronic graft-versus-host disease (cGVHD) is a severe and frequent complication of allogenic bone marrow transplantation which is often treated with extracorporeal photochemotherapy (ECP) with a positive clinical outcome in patients resistant to conventional protocols. The mechanism of action of ECP has not been fully elucidated, although several authors have reported that it is able to induce apoptosis. Using samples obtained from ten cGVHD patients, we sought to determine whether lymphocytes treated with ECP underwent apoptosis and, above all, the mechanisms involved. Lymphocytes at four stages were isolated: immediately before ECP, from the last buffy coat collected, after UV irradiation prior to reinfusion, and the day after ECP. When cultured for 48 h, lymphocytes treated with ECP underwent accelerated apoptosis (tested as annexin V binding cells and as intracellular histone-associated DNA fragments) in comparison with lymphocytes from the other samples. This enhanced programmed cell death could not be prevented by IL-2. Immediately after isolation, there was no difference in Bcl 2 or bax expression among the four different samples, or in Fas and FasL mRNA. However, when cultured, lymphocytes treated with ECP showed a rapid downregulation of Bcl-2, an upregulation of bax with an increased bax/Bcl-2 ratio, a decrease in bcl-2 mRNA and an increase in Fas. No changes were detectable in lymphocytes from the other samples. IL-2 and TNF-alpha production was not significantly different among lymphocytes from the four samples. In conclusion, in patients affected by cGVHD, ECP induced apoptosis of lymphocytes with the involvement of both the Fas/FasL system and the Bcl-2 protein family. PMID- 12883828 TI - The L266V tau mutation is associated with frontotemporal dementia and Pick-like 3R and 4R tauopathy. AB - We report a case of rapidly progressive frontotemporal dementia presenting at age 33 years. At autopsy there was severe atrophy of the frontal and temporal lobes. Tau-positive Pick bodies, which ultrastructurally were composed of straight filaments, were present, accompanied by severe neuronal loss and gliosis. RD3, a tau antibody specific for the three-repeat (3R) isoforms, labeled the Pick bodies. ET3, a four-repeat (4R) isoform-specific tau antibody, did not label Pick bodies, but highlighted rare astrocytes, and threads in white matter bundles in the corpus striatum. Analysis of the tau gene revealed an L266V mutation in exon 9. Analysis of brain tissue from this case revealed elevated levels of exon 10+ tau RNA and soluble 4R tau. However, both 3R and 4R isoforms were present in sarkosyl-insoluble tau fractions with a predominance of the shortest 3R isoform. The L266V mutation is associated with decreased rate and extent of tau-induced microtubule assembly, and a 3R isoform-specific increase in tau self assembly as measured by an in vitro assay. Combined, these data indicate that L266V is a pathogenic tau mutation that is associated with Pick-like pathology. In addition, the results of the RD3 and ET3 immunostains clearly explain for the first time the presence of both 3R and 4R tau isoforms in preparations of insoluble tau from some Pick's disease cases. PMID- 12883829 TI - Familial British dementia: colocalization of furin and ABri amyloid. AB - Familial British dementia (FBD) is an autosomal dominant condition caused by a point mutation in the stop codon of the BRI gene. This mutation extends the normal precursor protein (PP) of 266 amino acids to the next stop codon, which is at amino acid 277. Kim and colleagues demonstrated in vitro that furin can process both the normal protein BriPP and the extended protein ABriPP to produce C-terminal fragments of 23 and 34 amino acids. The abnormal C-terminal fragment, ABri, accumulates in FBD in the form of extracellular amyloid deposits. The objective of our study was to determine if furin is associated with ABri in FBD. Brain tissue of one case of FBD, four cases of Alzheimer's disease (AD) and two controls were studied by immunohistochemistry using antibodies against furin, beta-amyloid protein and ABri. In FBD, furin was found to be colocalized with ABri deposits and amyloid angiopathy in all areas examined. In contrast beta amyloid deposits in AD were not immunostained by the furin antibody. In normal as well as pathological cases, clusters of neurons in the hippocampus and neocortex showed light to moderate furin immunostaining, while peptidergic neurons of the hypothalamus showed intense furin-immunostaining. These data suggest that furin may be involved in producing the pathological fragment of ABriPP in vivo and that inhibition of furin might be a method of treating this disorder. PMID- 12883831 TI - Remodeling of cardiomyocyte ion channels in human atrial fibrillation. AB - This review is focused on electrical adaptational processes in patients with chronic AF. Cellular electrical remodeling includes shortening of action potential duration and effective refractory period that can be explained by concomitant alterations in ion channel activity. While most currents studied are reduced or unaffected, the inward rectifier I(K1) is increased in amplitude. The time courses of these changes and the putative molecular mechanisms suggest that electrical remodeling in chronically fibrillating human atria are adaptive processes. New therapeutic options could consist of supporting rather than reversing the adaptive mechanisms. PMID- 12883830 TI - Variability and heterogeneity in Alzheimer's disease with cotton wool plaques: a clinicopathological study of four autopsy cases. AB - We describe three cases of early- (cases 1-3, 28-39 years) and one of late-onset (case 4, 76 years) Alzheimer's disease (AD) with 'cotton wool' plaques (CWPs) but without a family history indicating autosomal dominant inheritance. The early onset cases, but not the late-onset case, showed remarkable aggression, disinhibition, and impulsiveness. Spastic paraparesis was observed in only one early-onset case. Hematoxylin-eosin-stained sections showed numerous CWPs, especially in the temporal cortex, in all cases. Bielschowsky-stained sections showed neurofibrillary tangles and minor neuritic changes surrounding the CWPs in three cases, but not in case 2. Gallyas-Braak-stained sections showed weak argyrophilia in homogeneous material of the CWPs in cases 2 and 4. Quantitative analysis demonstrated that Abeta42 was deposited more predominantly than Abeta40 in three cases. However, in case 2, approximately twice as much Abeta40 as Abeta42 was deposited. Tau immunostaining demonstrated neuritic changes in three cases, but not in case 2. alpha-Synuclein-positive Lewy bodies (LBs) and astrocytic lesions containing non-Abeta component of AD amyloid (NAC), a central fragment of alpha-synuclein, were found in case 3. In conclusion, (1) a frontal lobe syndrome-like personality change may be one of the characteristic clinical features of early-onset CWP-AD, (2) the deposition pattern of Abeta40 and Abeta42 in CWP-AD is more variable than that of presenilin-1-linked cases, (3) Abeta deposition can result in development of dementia without tau pathology, and (4) CWP-AD with LBs and several other neurodegenerative disorders with LBs share a common process involving alpha-synuclein and NAC deposition. PMID- 12883832 TI - Cardioprotection 'outside the box'--the evolving paradigm of remote preconditioning. AB - Conventional ischemic preconditioning is the phenomenon whereby brief episodes of myocardial ischemia render the ischemic territory resistant to a subsequent, sustained ischemic insult. A growing body of evidence further indicates that brief ischemia applied in distant organs and tissues can also protect naive, virgin myocardium from ischemic injury. In this review, we describe the initial observations that provided the impetus for the study of 'remote preconditioning', and summarize our current knowledge of the three facets of 'preconditioning at a distance' --intra-cardiac, inter-organ and transferred inter-cardiac preconditioning. PMID- 12883833 TI - Involvement of p38 MAPK and JNK in heat stress-induced cardioprotection. AB - The present study investigated whether heat stress-induced cardioprotection involves alterations in the pattern of p38 mitogen activated protein kinase (p38MAPK) and c-Jun NH2 - terminal kinase (JNK) activation during ischaemia - reperfusion in a model of isolated perfused rat heart. Wistar rats were subjected to whole-body hyperthermia at 42 degrees C for 15 min (HS), while untreated animals served as controls (CON). Twenty four hours later, CON and HS isolated hearts were perfused in a Langendorff mode and subjected to 20 min of zero-.ow global ischaemia followed by 45 min of reperfusion. Postischaemic recovery of left ventricular developed pressure at 45 min of reperfusion was expressed as % of the initial value (LVDP%). Activation of p38 MAPK and JNK was assessed by standard Western blotting techniques using a dual phospho-p38 MAPK and phospho p46 JNK and p54 JNK antibodies. The levels of phospho-p38 MAPK at the end of reperfusion were not different in HS as compared to CON hearts. The levels of phospho-p46 JNK and p54 JNK were 1.4- and 1.6-fold less in HS than in CON hearts respectively, p < 0.05. LVDP% was 60.3 (s.e.m., 6.3) for HS and 42.9 (4.1) for CON, p < 0.05. In summary, heat stress pretreatment improves postischaemic recovery of function in isolated rat hearts and this is associated with suppressed JNK activation in response to ischaemia-reperfusion. PMID- 12883834 TI - Selective modulation of endogenous nitric oxide formation in ischemia/reperfusion injury in isolated rat hearts--effects on regional myocardial flow and enzyme release. AB - The role of nitric oxide (NO) in ischemia/reperfusion injury is controversial. We tested the role of inducible NOS (iNOS) in the ischemia/reperfusion injury in isolated rat hearts using the selective iNOS inhibitor S-methylisothiourea sulfate (SMT) and the non-selective NOS inhibitor N(G)-nitro-L-arginine methyl ester (L-NAME). After 15 min of stabilization in Langendorff mode, hearts were perfused either with normal Krebs-Henseleit buffer, buffer containing 100 microM L-NAME, 0.5 microM SMT or 50 microM SMT for 5 min and were subjected to 25 min of ischemia followed by 30 min of reperfusion. Left ventricular developed pressure (LVDP) and total coronary flow (CF) were recorded continuously. After ischemia/reperfusion, a marked expression of iNOS protein was demonstrated by Western blotting, while virtually no iNOS protein was present in hearts without ischemia/reperfusion. Regional myocardial blood flow (RMBF) was measured with colored microspheres. Coronary vasoactive concentration of L-NAME and SMT depressed myocardial function as shown by decreased LVDP, dP/dt(max) and coronary.ow before ischemia. After ischemia the recovery of the total CF was impaired in L-NAME and 50 microM SMT pretreated hearts which was related to homogenous RMBF decrease in the right and left ventricle compared to that in control group. Low concentration SMT (0.5 microM) showed no coronary vasoactive effects before ischemia and attenuated ischemia/reperfusion injury indicated by lower ischemic contracture at 25 min of ischemia and reduced CK and LDH release during reperfusion. Thus, NOS inhibition did not affect blood flow distribution in rat hearts either in the pre-ischemic or reperfusion period. Selective iNOS inhibition reduced ischemic injury by reducing ischemic contracture and CK as well as LDH release during reperfusion. PMID- 12883835 TI - Enhancement of carotid chemoreceptor reflex and cardiac chemosensitive reflex in the acute phase of myocardial infarction of the anesthetized rabbit. AB - In the acute phase of cardiac ischemia there is an imbalance of the autonomic outflow with a depression of the baroreceptor reflex. Carotid chemoreceptor stimulation evokes an increase on arterial blood pressure and bradycardia in the anesthetized and paralyzed animal. The activation of cardiac chemosensitive fibers elicit the Bezold-Jarisch reflex comprising a decrease of arterial blood pressure and bradycardia. In the present study, we studied the modifications of the carotid chemoreceptor reflex and the Bezold-Jarisch reflex elicited during the acute phase of myocardial infarction (MI) in the anesthetized and paralyzed rabbit. Rabbits were anesthetized with pentobarbitone, paralyzed and artificially ventilated. The carotid sinus region was exposed and a cannula was inserted retrogradely through the external carotid artery into the carotid bifurcation; the carotid body was stimulated by a lobeline injection. A catheter was advanced, via the right carotid artery, to the origin of the aorta and the Bezold-Jarisch reflex was evoked by an injection of ATP. The baroreflex was provoked by an increase in after-load or by clamping the common carotid artery. Heart ischemia was provoked by ligation of the descending coronary artery. Arterial blood pressure, carotid artery pressure, heart rate and electrocardiogram were monitored. Stimulation of cardiovascular and cardiac receptors was performed before and after coronary ligation. Results show an overall increase in the cardiovascular reflex responses elicited by stimulation of chemically activated receptors and an overall decrease of the baroreceptor responses after MI. In conclusion, these data show the existence of an enhancement of the reflex cardiovascular responses to carotid chemoreceptor and cardiac chemosensitive receptors stimulation and confirmed the depression of baroreceptor reflexes following heart ischemia that could account for the imbalance of the autonomic output observed in the acute phase of myocardial infarction. PMID- 12883836 TI - [The impact factor or the fairy tale of impartiality]. PMID- 12883837 TI - Anticoagulation with the low-molecular-weight heparin dalteparin (Fragmin) in atrial fibrillation and TEE-guided cardioversion. AB - BACKGROUND: Cardioversion (CV) in atrial fibrillation can cause arterial embolism. Effective anticoagulation clearly reduces the risk. In practice, in every third case anticoagulation is not in line with the recommendations. Simplification can be achieved, and time gained, by transesophageal echocardiography (TEE) due to the shorter anticoagulation period prior to CV, and by use of low-molecular-weight heparin (LMWH) for anticoagulation. As yet little data is available on LMWH in cardioversion. The aim of this cohort study was to investigate the administration of a LMWH in this indication under everyday clinical conditions. METHODS: 125 patients treated as inpatients for atrial fibrillation or -flutter received the LMWH Fragmin (dalteparin 2 x 100 anti-Xa units/kg, maximum dosage 2 x 10,000 anti-Xa units subcutaneously). In the presence of a relevant indication, TEE-guided CV was performed. The application of dalteparin was terminated as soon as effective anticoagulation had been achieved from phenprocoumon or once anticoagulation was no longer indicated. RESULTS: 125 patients with atrial fibrillation or -flutter received dalteparin for a median of 11 days (range of 3-41 days). TEE was performed in 39 patients. Five patients revealed a thrombus in the left atrial appendage in the TEE, and one patient died from suspected cerebral embolism over the further course. In the remaining 124 patients, no thromboembolic event was established. Successfully cardioverted were 26 of 34 patients (76%) who had no thrombus in the TEE. Serious adverse effects did not ensue. CONCLUSION: Simple, well tolerated and effective anticoagulation is possible with dalteparin in TEE-guided CV. Due to the methodic limitations of a cohort study and the low incidence of emboli, the efficacy of dalteparin in this indication needs to be further confirmed by prospective and randomized studies. PMID- 12883838 TI - Prediction of early recurrence of atrial fibrillation after external cardioversion by means of P wave signal-averaged electrocardiogram. AB - BACKGROUND: Therapy of atrial fibrillation by electrical cardioversion (CV) is limited by the high rate of recurrences. Early recurrence of atrial fibrillation (ERAF) occurs in a subgroup of patients whose characteristics are poorly defined. This prospective study was performed to evaluate if the P wave signal-averaged ECG (PSAECG) is able to identify patients with an increased risk of ERAF after CV. METHODS: Patients with an indication for elective external CV were enrolled. After successful CV, PSAECGs were recorded at 0.5, 1, 24 h and 1 week. The ability of PSAECG parameters (signal-averaged P wave duration, PWD; root-mean square of the voltage of the terminal 20, 30, and 40 ms of the signal-averaged P wave; RMS20, RMS30, RMS40) to predict ERAF (prospectively defined as AF recurrence within 4 h after CV) was assessed. RESULTS: Of 111 consecutive patients, 7 experienced ERAF, 30 patients had AF recurrence later during the 1 week follow-up. Patients with ERAF had a significantly prolonged signal-averaged PWD compared to patients who remained in SR (194+/-16 ms vs 139+/-3 ms at 0.5 h, p<0.001). As ROC analysis revealed, a PWD >/=154 ms at 30 min after CV had the highest predictive accuracy for ERAF (sensitivity 100%, specificity 82%, positive predictive value 33%, negative predictive value 100%). Other parameters of the PSAECG did not reveal significant differences between patients with and without ERAF. CONCLUSIONS: The PSAECG provides a sensitive noninvasive tool for detection of patients at risk of ERAF. Thus identified, tailored pharmacological therapy is conceivable to prevent ERAF. PMID- 12883839 TI - [Short-term heart rate turbulence analysis versus variability and baroreceptor sensitivity in patients with dilated cardiomyopathy]. AB - New methods for the analysis of arrhythmias and their hemodynamic consequences have been applied in risk stratification, particularly to patients after myocardial infarction. This study investigates the suitability of shortterm heart rate turbulence (HRT) in comparison to heart rate and blood pressure variability as well as baroreceptor sensitivity analyses to characterize the regulatory differences in patients with dilated cardiomyopathy (DCM) and healthy controls. In this study, 30 minutes data from noninvasive continuous blood pressure and ECG of 37 DCM patients and 167 controls under standard resting conditions were analyzed. The results showed highly significant differences between DCM patients and controls in heart rate and blood pressure variability as well as baroreceptor sensitivity parameters. Applying a combined heart rate-blood pressure trigger, in 24.3% (9) of the DCM patients and in 11.3% (19) of the controls ventricular premature beats were detected. This fact demonstrates the constricted applicability of short-term HRT analyses. However, the HRT parameters showed significant differences in this subgroup with ventricular premature beats (Turbulence Onset: DCM: 1.80+/-2.72, CONTROLS: -4.34+/-3.10, p<0.001; Turbulence Slope: DCM: 6.75+/-5.50, CONTROLS: 21.30+/-17.72, p = 0.021). Considering all (including HRT) parameters in the subgroup with ventricular beats, a discrimination rate between DCM patients and controls of 88.0% was obtained (max. 6 parameters). In comparison, in the total group this rate was 86.3% (without HRT parameters). The comparable classification rates and the high correlations between heart rate turbulence and variability and baroreflex parameters point to a more universal applicability of the last-mentioned methods. PMID- 12883840 TI - Chemoreflexsensitivity among patients with paroxysmal atrial fibrillation. AB - Atrial fibrillation is the most common cardiac arrhythmia with typical complications of thromboembolisms. The autonomic nervous system is an important factor for the initiation of arrhythmias. A vagally or adrenergically hyperfunction could cause the initiation of paroxysmal atrial fibrillation (PAF). METHOD: We measured the chemoreflexsensitivity (CHRS) among 110 patients to determine a disturbed autonomic function as risk factor for PAF. We examined 45 patients with PAF (group A), 45 patients with sinus rhythm (group B) and 20 young volunteers (group C). The ratio between the difference of RR intervals in ECG and venous pO(2) was measured for the determination of CHRS. The margin of the CHRS was 3 ms/mmHg. RESULTS: Patients of group A had a significantly lower CHRS compared to group B (1.56+/-1.46 vs 6.29+/-3.71 ms/mmHg, p<0.0008) or group C (1.56+/-1.46 vs 6.35+/-4.29 ms/ mmHg, p<0.0003). A significant difference between group B and C could not be observed (6.29+/-3.71 vs. 6.35+/-4.29 ms/mmHg, p = n.s.). A specificity of 74% and a sensitivity of 71% was achieved for identifying patients with PAF by using a margin of 3 ms/mmHg for the CHRS. CONCLUSIONS: An analysis of CHRS seems to be an appropriate method to demonstrate a neurovegetative imbalance which might be one possible trigger mechanism of PAF. The predictive power has to be examined by prospective investigations of a larger patient population. PMID- 12883841 TI - A prospective study on ultrasound-guided compression therapy or thrombin injection for treatment of iatrogenic false aneurysms in patients receiving full dose anti-platelet therapy. AB - BACKGROUND: False aneurysms (FA) develop at the puncture site in up to 6% of percutaneous cardiovascular procedures. Previous management included surgery or manual compression. Recently, selective injection of thrombin has been proposed as an alternative. However, there has been no direct comparison of thrombin injection to manual compression. AIM: To study the effectiveness of manual compression compared to that of thrombin injection in patients with false aneurysms on full-dose aspirin and clopidogrel. METHODS AND PROTOCOL: All patients with a clinically suspected FA after percutaneous invasive procedures were recruited for the study. The patients were examined with color ultrasound (7.5 MHz transducer). The minimum and maximum diameters of the false aneurysm and the distance between the surface and the false aneurysm were measured online. Under local anesthesia, manual compression was applied under sonographic guidance in all patients. If compression stopped flow into the false aneurysm, manual compression was applied for a maximum of 40 min followed by compression bandage for a minimum of 12 hours. If compression failed, thrombin was injected under ultrasound guidance. RESULTS: Thirty-six patients had a FA. Their age ranged from 58 to 90 years (mean 71+/-9 years). All patients were taking aspirin (median dose 100 mg per day) and clopidogrel (median dose 75 mg per day). Additionally, 24 patients had received subcutaneous heparin (7500 to 12 500 units) or enoxaprin (0.4-1.0 ml) 3 to 12 hours before treatment. The mean width of the false aneurysm was 22.1+/-3 mm, mean length 33.6+/-35.4 mm, and mean depth 19.5+/-8.2 mm. In six patients (17%), ultrasound-guided manual compression was tolerated, succeeding after 5 to 31 minutes. Thirty patients received thrombin injections (100-1800 units, mean 880+/-470 units, median 800 units). Complete thrombosis occurred in 28 patients (93%). Surgery was performed in the other two patients. The thrombin injection was not associated with any complications. In particular, there were no peripheral vascular complications. CONCLUSION: In patients with FA taking aspirin and clopidogrel, selective thrombin injection is more effective than manual compression. PMID- 12883842 TI - [Documentation in the cardiac catheterization laboratory using electronic databases--experiences in 176 German cath labs]. AB - Due to the introduction of a variety of new features to the German health system, such as a new reimbursement system for inpatient work based on diagnosis-related groups, an obligation to detailed documentation for total quality management, as well as due to the continuing increase in the amount of imaging and other data within invasive cardiology, the use of electronic databases for documentation in the cardiac cath lab is of significant importance. The user friendliness and robustness of the software system for daily clinical practice is crucial for the acceptance of electronic documentation systems by physicians, nurses and technicians. This survey was aimed to explore the current practice and experiences of German cardiac catheterization labs with electronic databases particularly designed for documentation in invasive cardiology, including report generation of cath studies, risk and complication management and logistics for medical material ordering for the cath lab. Of the 176 hospitals and practices having agreed in taking part in this surveillance 165, are currently using an electronic database for administration of medical data and study reports. In 64% of the cath labs, a sub-specialized software system for detailed documentation of the course and the results of an invasive procedure, the human and material resources used, generation of the cath report, and documentation of procedural and post-procedural complications for total quality management is used. The acceptance and satisfaction with the software solutions currently available reveals to be rather inhomogeneous. While user-friendliness and flexibility of a number of software products were positively mentioned, main issues, such as missing hardware of software interfaces with the hospital information system for patient administration, missing stability of several systems, image management too complex and report configuration too inconvenient for daily clinical practice, were criticized. In general, in the light of continuing increase of the amount of clinical and imaging data as well as due to the legal obligation of detailed procedural documentation in all areas of invasive cardiology, a documentation system based on electronic databases bears the potential of an enormous economization of the daily workload of a cardiac cath lab. Prerequisites for the acceptance of such a system in clinical practice is not only the stability and reliability of the system but also the ability to link such a sub system to existing databases via suitable interfaces and the flexibility of the system for modification of its features to meet the individual requirements of each user. PMID- 12883843 TI - [Evaluating the first German diagnosis-related groups (G-DRG) in cardiological patients: problems in the correct medical and economic grouping]. AB - About three years ago, the German Government initiated a complete change in the reimbursement system for costs of the in-hospital treatment of patients. A commission of representatives from every component of the German health system decided to adapt the Australian refined Diagnosis Related Groups (AR-DRG system). The AR-DRG system was selected as it would fit best to the German system and because of its high flexibility and preciseness reflecting severity of diseases and treatments. In October 2002, the first German Diagnosis Related Groups (G DRGs) were calculated from the data of about 116 hospitals. These data now allow first analyses in how far a correct and precise grouping of patients in specific hospital settings is indeed performed and corresponds to the actual costs. Thus, we thoroughly calculated all costs for material and personnel during the in hospital stay for each patient discharged during the first 4 months of 2002 from our cardiological department. After performing the grouping procedure for each patient, we analyzed in how far inhomogeneous patient distribution in the DRGs occurred and which impact this had on costs and potential reimbursements. Several different problems were identified which should be outlined in this work regarding three G-DRGs: costs of patients who received an implantable cardioverter defibrillator (F01Z) were markedly influenced by multimorbidity and additional expensive interventions which were not reflected by this G-DRG. Use of numerous catheters and expensive drugs represented a major factor for costs in patients with coronary angioplasty in acute myocardial infarction (F10Z) but seemed to be not sufficiently included in the cost weight. A specific area of patient management in our department is high frequency ablation of tachyarrhythmias which is included in other percutaneous interventions (F19Z). Complex procedures such as ablation of ventricular tachycardia or new innovative procedures as ablation of atrial fibrillation were associated with high costs leading to inadequate reimbursement. Furthermore, problems in the associated codes for diseases and procedures became apparent. Opportunities for future optimization such as specific new DRGs, splitting of DRGs, or the impact of changes in reimbursement for high-outliers were discussed. PMID- 12883844 TI - [A "late" scimitar syndrome. Diagnostic contribution of cardiac computed tomography]. AB - A 51-year-old female hospitalized with a non-specific colitis, presented a crescent-like shadow in the right lower lung accompanied by a reduced right lung volume on a routine chest x-ray. There was no family history of congenital heart disease. The initially performed, noninvasive, contrast enhanced cardiac CT (electron-beam tomography [EBT]) proved the suspected diagnosis of a partial, anomalous pulmonary, transdiaphragmatic vein drainage (APVD) in combination with a hypoplastic right lower lobe and dextrocardia. These findings are in accordance with scimitar syndrome. Regarding to the clinical situation with symptoms like slowly progressive dyspnea on exertion and low exercise tolerance for the last 2 years and an invasively documented left-to-right shunt ratio >50% (Qp:Qs = 2.6 : 1) surgical repair was recommended. The anomalous vein was connected to the left atrium creating a "neo-septum". On a postoperative checkup after 9 months the patient is without any medication, symptoms during moderate activity are relieved, exercise tolerance was substantially better and noninvasive imaging visualized the corrected drainage of the anomalous vein to the left atrium. PMID- 12883845 TI - [Spontaneous pneumomediastinum as a cause of chest pain]. AB - Two patients, both 16 years old, presented because of chest pain after extended sports activity. The thoracal X-ray showed trapped air in the mediastinum especially around the cervical vessels in one patient. In the thoracal X-ray of the other patient a vertical lucent streak along the left side of the heart, showing the pleura as a fine opaque line, was found. Both developed subcutaneous emphysema after a few hours, which led to the diagnosis spontaneous pneumomediastinum. The chest pain resolved under analgetic medication, and after resorption of the subcutaneous emphysema both patients recovered completely. The main differential diagnosis of the spontaneous pneumomediastinum is besides pericarditis and myocarditis, the spontaneous esophageal perforation (also called Boerhaave syndrome), with the high morbidity, it has to be ruled out consequently. PMID- 12883846 TI - [Position paper on the use of drug eluting stents (DES) in percutaneous coronary interventions in patients with coronary heart disease]. PMID- 12883850 TI - Adrenocortical tumors in childhood. AB - Adrenocortical tumors (ACT) are rare in childhood. Determination of malignant potential is difficult. To assess the presentation, clinical behaviour, and histology of these tumors in our center, the records of nine patients with ACTs between 1989 and 2000 were analyzed. The age, sex, clinical presentation, investigations, treatment, follow-up data, and current status were determined. The pathological features were reanalyzed by our pathologist. The endocrine profile of the tumors was also retrieved from the records. There were two males and seven females; the median age at referral was 2.5 years. Two patients (one male, one female) presented with clinical features of Cushing's syndrome with hypertension. Four girls had features of Cushing's syndrome with associated virilization. One 11-year-old girl presented with hypertension, hypokalemia, and pseudoparalysis due to an aldosterone-secreting tumor. One girl presented with precocious pubic hair and clitoromegaly. One male had evidence of true precocious puberty. The endocrine profile was available in eight cases: cortisol levels were raised in four, testosterone in four, and aldosterone in one. One boy had pubertal levels of follicle-stimulating hormone and testosterone. Surgery was the only modality of therapy used. Histopathology showed capsular invasion in four patients and vascular invasion in two. The median tumor weight was 80 g (15-550 g) and the size ranged from 1.5 x 1.5 to 12 x 12 cm. Five patients are alive without tumor recurrence at a median follow-up time of 2 years. Tumor recurrence was seen in two patients, one with capsular rupture during surgery and another with a tumor weight of 16 g with associated capsular and vascular invasion. Atypical modes of presentation like pseudoparalysis must be kept in mind. Histologic criteria for malignancy in ACT are unreliable. Tumor weight may not always be accurate in predicting tumor behaviour. Complete surgical excision remains the only effective and potentially curative treatment. PMID- 12883851 TI - Wound infections in pediatric surgery: a study of 575 patients in a university hospital. AB - Surgical wound infections (WI) remain a significant source of postoperative morbidity. This epidemiologic study was undertaken to determine retrospectively the incidence of postoperative WI in children in a university hospital and include critical comparisons of pediatric surgery WI rates between different international reports. As few data exist on postoperative WIs in pediatric patients, in contrast to numerous reports in adults, all infants and children undergoing operations in the pediatric surgical service in our institution during a 7-year period were reviewed for development of a WI, a total of 537 patients who underwent 575 operations. WIs occurred in 39 cases (6.7%). Clean wounds (56.8% of patients) had an infection rate of 2.7%, clean-contaminated (23.1%) 10.5%, contaminated (12.9%) 13.5%, and dirty/infected (7.2%) 14.6%. Increasing duration of operation ( P < 0.001), contamination at operation ( P < 0.001), and a new element in the operation - a resident or intern - ( P < 0.001) were all associated with a higher incidence of infection, despite efforts at infection control practices including improved sterilization methods and barriers, surgical technique, and availability of antimicrobial prophylaxis. The total incidence of wound infection in this population was comparable to that in other reports. Comparing children who developed a wound infection with those who did not, there were no significant differences in age, sex, American Society of Anesthesiologists preoperative assessment score, length of preoperative hospitalization, location of operation (intensive care unit vs operating room), the presence of a coexisting disease or remote infection, or the use of perioperative antibiotics. These baseline data may aid in forming strategies to lower the risk of WI in children. Our results suggest that WIs in children are related more to factors at operation than to the patients overall physiologic status. PMID- 12883852 TI - Reverse serosal demucosalized gastrocystoplasty. AB - A novel technique of augmentation cystoplasty is suggested. The patch which is advocated is less likely to cause electrolyte imbalance, is autologous and easily available. It will avoid all the gut-mucosa-related early and late postoperative complications. PMID- 12883853 TI - Fetal cricotracheal manipulation: effects on airway healing, cricoid growth and lung development. AB - The last decade has seen significant advance in the surgical management of pediatric subglottic stenosis, which remains one of the most fascinating problems of the laryngotracheal complex (LTC). Refined techniques for operating on these fragile structures should reduce cricotracheal scarring to a minimum, thus avoiding a lot of severe postoperative complications in a tricky moment of laryngeal's growing up. Experimental works indicates that the LTC growth is variously affected by longitudinal anterior, posterior or lateral incisions and actually the indications for laringotracheoplasty or cricotracheal resection in children with subglottic stenosis are still unclear. Reports on fetal manipulation of cricotracheal tissues are lacking as well as early effects on airway healing, LTC growth and lung development. The aim of this study was to evaluate if the airway mucosal healing is regenerative and scarless after cricotracheal manipulation in fetuses of New Zealand White Rabbits (NZWRFs). The consequences of fetal incisions on the cricoid growth and lung development are also examined, in a group of 12 NZWRFs, manipulated at 25+/-1 days of gestational age. The does underwent halothane anesthesia and all received a bilateral longitudinal cricoidotracheotomy. Twenty sham-operated fetuses were submitted to a limited cervicotomy (control's group). At the time of retrieval (31+/-0.5 days), en bloc laryngotracheobronchial tree and lungs were collected and processed for histological and morphometric analysis. Parameters recorded included: 1) histological full-thickness examinations focusing on inflammation, foreign body reaction, fibrosis, neochondrogenesis; 2) morphometric analysis, including the fetal Subglottic Diameter (FSD), the fetal Subglottic Area (FSA), the Radial Alveolar Count (rAC) and Computer Assisted Morphometric Colorimetry (CAmc); 3)analysis of lung hypoplasia (LH) by means of lung weight/body weight (LW/BW) ratio, protein and DNA indexes; 4) finally, different fractions of lung tissue phospholipids for lung maturity assessment were studied. Student's t test, when indicated, was performed for statistical analysis (p <0.05 = significant). There was no maternal mortality in this study. Ten fetuses were available for a final evaluation (16.6% mortality). In one case only, an incomplete closure of the fetal cricoidotomy was seen and could be probably due to a technical mistake. Mean fetal subglottic diameter and area were respectively 0.13+/-0.05 mm and 3.15+/-0.45 mm2 in both groups. As well as in fetal dermal repair, regeneration of the airway cartilage and mucosa were complete and scarless. LW/BW ratio, DNA content and analysis of different fractions of phospholipids were similar in experimental vs. the control group. These findings suggest that the healing processes were fibrosis-free and without evidence of scars. A complete closure of the incisions was achieved without stenosis of the fetal subglottic region. In addition, it seems that the fetal cricoidotracheotomy doesn't interfere with the laryngeal function which coordinate the amount of liquid leaving the lungs via the trachea. In addition, only a small leakage of amniotic fluid is shown and this could be responsible for normal and mature lungs. PMID- 12883854 TI - Simple method for in vitro bladder urothelium regeneration on a heterologous acellular matrix. PMID- 12883855 TI - The utility of peritoneal drains in children with uncomplicated perforated appendicitis. AB - Peritoneal drainage in children with uncomplicated perforated appendicitis (UPA) is still controversial. Many pediatric surgeons prefer not to drain the peritoneal cavity in such cases. However, there is no randomized controlled study performed in children. We aimed to study the effects of peritoneal drainage in children with UPA in a randomized prospective trial. One hundred and forty consecutive patients with UPA were divided randomly into 2 groups. Group I (70 patients) consisted of cases with peritoneal drainage, and group II (70 patients) without drainage. UPA is defined as perforated appendicitis with no more discoloration of peritoneal fluid after peritoneal wash out. Cases with localized abscess in the peritoneum were excluded from the study. In all patients, the ages, duration of symptoms, nasogastric drainage and hospitalization, and complications after surgery were recorded. The duration of hospitalization and nasogastric draining time were significantly lower in patients without peritoneal drainage. There was no difference in postoperative complications between the two groups. The onset of oral intake after surgery was significantly earlier in group II patients. Placing drains in the peritoneum does not improve outcome in UPA. Therefore, we do not recommend routine drainage of children with UPA. PMID- 12883856 TI - Scrotoshisis, bilateral extra-corporeal testicular ectopia and testicular torsion. AB - Although anomalies of testicular descent are common, scrotoschisis and extracorporeal testicular ectopia is rare with only 3 reported cases in literature. This is a report of a neonate presenting with scrotoschisis and bilateral extracorporeal testicular ectopia. The right testis underwent torsion before presentation, resulting in gangrene of that testis, requiring orchidectomy. The scrotum was explored and the viable left testis placed in the scrotum and fixed in that position. The left testis has remained normal at 9 months of follow up. The possible embryology and management of this curious anomaly is discussed. PMID- 12883858 TI - The Freter model: a simple model of biofilm formation. AB - A simple, conceptual model of biofilm formation, due to R. Freter et al. (1983), is studied analytically and numerically in both CSTR and PFR. Two steady state regimes are identified, namely, the complete washout of the microbes from the reactor and the successful colonization of both the wall and bulk fluid. One of these is stable for any particular set of parameter values and sharp and explicit conditions are given for the stability of each. The effects of adding an anti microbial agent to the CSTR are examined. PMID- 12883857 TI - A mathematical model for reconstitution of granulopoiesis after high dose chemotherapy with autologous stem cell transplantation. AB - High dose chemotherapy supported with hematopoietic progenitor cells gives a characteristic neutropenic period (blood neutrophils < 0.5 x 10(9) c/l) ranging from 10 to 16 days. The question of a correlation between the CFU-GM content of the transplanted CD34+ cells and time to neutrophil recovery by patients having been given high-dose chemotherapy (HD-CT) with stem cell support was addressed by means of a mathematical model of granulopoiesis. The model utilizes a convection reaction partial differential equation (PDE) with feedback from a cytokine compartment on proliferation, maturation, and mobilization of granulocytes from bone marrow to blood. The observed number of CFU-GM cells in the transplanted CD34+ cell autograft was used as input to the model. Using this approach, the observed gross relationship between CFU-GM content in the reinfused blood product and engraftment time could be reproduced. At the same time, the effects of assumed physiological mechanisms, especially some of the effects of G-CSF on proliferation rate, maturation rate, mobilization, and cell death, could be investigated and discussed relative to observed engraftment. The model makes it possible to explain how cytokines interfere with progenitor cell mobilization from bone marrow to blood, and it points out the implications of a regulating mechanism for the granulocyte maturation rate. PMID- 12883859 TI - Competitive exclusion and coexistence for pathogens in an epidemic model with variable population size. AB - We study an SIR epidemic model with a variable host population size. We prove that if the model parameters satisfy certain inequalities then competition between n pathogens for a single host leads to exclusion of all pathogens except the one with the largest basic reproduction number. It is shown that a knowledge of the basic reproduction numbers is necessary but not sufficient for determining competitive exclusion. Numerical results illustrate that these inequalities are sufficient but not necessary for competitive exclusion to occur. In addition, an example is given which shows that if such inequalities are not satisfied then coexistence may occur. PMID- 12883860 TI - Fractal dynamics in circadian cardiac time series of corticotropin-releasing factor receptor subtype-2 deficient mice. AB - Non-linear fractal analysis of circadian 24 hr heartbeat interval time series was performed in corticotropin releasing factor receptor-subtype 2 (CRFR2) deficient mice. We hypothesized that, as a result of its central as well as its peripheral expression, CRFR2 would mediate or interfere with the circadian rhythmicity. The dynamical properties of cardiac interbeat intervals were expected to be different between CRFR2 (+/+) and CRFR2 (-/-) mice when studied over an extended circadian 24 hr cycle. The dynamics of neurocardiac control were found to remain remarkably stable throughout the circadian cycle. In disagreement with the initial hypothesis, the dynamical properties underlying the cardiac control process were common to both CRFR2 (+/+) and CRFR2 (-/-) mice suggesting that control of heart rate does not rely on the elaborate interaction of the CRFR2-sensor and its intrinsic feedback arrangement. Lack of expression of CRFR2 would not compromise cardiac control and its dynamical output or is subserved by other, unknown mechanisms. Functional integrity of CRFR2 would not constitute an indispensable requirement of physiologic cardiac control. The circadian rhythm of heart rate is generated centrally and is independent of expression of CRFR2. While 'normal' strain C57BL/6N mice exhibit a circadian dark/light cycle of heart rate, absence of circadian fluctuations in transgenic CRFR2-mice (both +/+ and -/-) and 'normal' strain C57BL/6J mice points at the importance of other deficiencies that may be related to a common genetic background. Mutant mice that share a common 129SvJ- or C57BL/6J-derived genetic background may not present an optimal model for physiological studies of cardiovascular control. PMID- 12883861 TI - Enhancement of taxol production and release in Taxus chinensis cell cultures by ultrasound, methyl jasmonate and in situ solvent extraction. AB - This study evaluates the use of a novel mechanical stimulus, ultrasound (US), and a putative chemical elicitor, methyl jasmonate (MJ), combined with in situ solvent extraction (two-phase culture), to enhance taxol production by Taxus chinensis cells in suspension culture. The volumetric taxol yield was increased 1.5- to 1.8-fold with 2 min US treatment once or twice during a 4-week culture period, about 5-fold with 60-120 microM MJ, and 7- to 9-fold by in situ solvent extraction of taxol with dibutyl phthalate (DBP) (11% v/v). The percent of extracellular taxol or taxol release was also significantly increased. The combined use of US (day 5 or 9) and MJ treatment (day 7) resulted in taxol yields 20-50% higher than each of the treatments used alone. The most favorable strategy for taxol production was the application of US or MJ treatment, followed by in situ solvent extraction, giving rise to a taxol yield of 33-35 mg/l, about 17 fold higher than the control, at 1.9 mg/l. It was found that the organic solvent DBP, as well as US and MJ, stimulated the enzyme activity of secondary metabolic pathways, which was partially responsible for the enhanced taxol production. PMID- 12883862 TI - Influence of growth conditions on bacteriocin production by Brevibacterium linens. AB - The influence of temperature, NaCl concentration and cheese whey media on growth of Brevibacterium linens ATCC 9175 and production of bacteriocin-like antimicrobial activity was studied. Bacteriocin production and activity were higher at 25 degrees C than at 30 degrees C. No significant growth or production of bacteriocins was observed at 37 degrees C. When bacteriocin production was investigated in media containing different concentrations of NaCl, increased activity was observed in media containing 40 or 80 g l(-1), but not 120 g l(-1) NaCl. The addition of NaCl resulted in a significant increase in specific production rates of bacteriocin-like activity. Antimicrobial activity was also observed by cultivation of B. linens at 25 degrees C in cheese whey media. PMID- 12883863 TI - Gene cloning, expression, and crystallization of a thermostable exo-inulinase from Geobacillus stearothermophilus KP1289. AB - The gene ( inuA) encoding exo-inulinase (EC 3.2.1.80) was cloned from the thermophilic Geobacillus stearothermophilus ( Bacillus stearothermophilus) KP 1289 growing at between 41 degrees C and 69 degrees C. The inuA gene consisted of 1,482 bp encoding a protein of 493 amino acids. The deduced polypeptide of molecular mass ( M) 56,744 Da showed strong sequence similarity to Pseudomonas mucidolens exo-inulinase, Bacillus subtilis levanase, Paenibacillus polymyxa ( Bacillus polymyxa) fructosyltransferase, and so on, indicating that the enzyme belonged to glycosyl hydrolase family 32. The M of the purified exo-inulinase, expressed in Escherichia coli HB101, was estimated as approximately 54,000 Da by both SDS-PAGE and gel filtration. These results suggested that the active form of the enzyme is a monomer. The enzyme was active between 30 and 75 degrees C with an optimum at 60 degrees C. The properties were identical to those of the native enzyme. Additionally, for the first time for a prokaryotic GH32 protein, crystals of the recombinant enzyme were obtained. PMID- 12883864 TI - Molecular, biochemical and ecological characterisation of a bio-catalytic calcification reactor. AB - Bio-catalytic calcification (BCC) reactors utilise microbial urea hydrolysis by autochthonous bacteria for the precipitation-removal of calcium, as calcite, from industrial wastewater. Due to the limited knowledge available concerning natural ureolytic microbial calcium carbonate (CaCO(3)) precipitation, the microbial ecology of BCC reactors has remained a black box to date. This paper characterises BCC reactor evolution from initialisation to optimisation over a 6 week period. Three key parameters were studied: (1) microbial evolution, (2) the (bio)chemical CaCO(3) precipitation pathway, and (3) crystal nucleation site development. Six weeks were required to establish optimal reactor performance, which coincided with an increase in urease activity from an initial 7 mg urea l( 1) reactor h(-1) to about 100 mg urea l(-1) reactor h(-1). Urease activity in the optimal period was directly proportional to Ca(2+) removal, but urease gene diversity was seemingly limited to a single gene. Denaturing gradient gel electrophoresis of 16S rRNA genes revealed the dynamic evolution of the microbial community structure of the calcareous sludge, which was eventually dominated by a few species including Porphyromonas sp., Arcobacter sp. and Bacteroides sp. Epi fluorescence and scanning electron microscopy showed that the calcareous sludge was colonised with living bacteria, as well as the calcified remains of organisms. It appears that the precipitation event is localised in a micro environment, due to colonisation of crystal nucleation sites (calcareous sludge) by the precipitating organisms. PMID- 12883865 TI - cDNA cloning and functional expression of alpha-glucosidase from Mortierella alliacea. AB - We recently purified an alpha-glucosidase comprising 61-kDa and 31-kDa subunits from the fungus Mortierella alliacea and characterized its soluble starch hydrolyzing activity. Here, the cDNA coding for this enzyme was cloned, revealing that it encodes a single polypeptide of 1,053 amino acids, with a calculated molecular mass of 117 kDa. Comparison between the deduced amino acid sequence and the partial sequences of the purified enzyme suggested that an immature protein can be converted into the two subunits of mature enzyme by post-translational processing at least three cleavage sites. Heterologous expression of recombinant alpha-glucosidase in yeast gave rise to a significant increase in hydrolytic activity toward maltose and soluble starch, in both intracellular and extracellular fractions. Immunoblot analysis using antiserum against the alpha glucosidase revealed that the active enzyme expressed in yeast is also composed of two subunits. The yeast expression system provides a model suitable for investigating the polypeptide-processing event and structure-function relationship of the alpha-glucosidase with unique substrate specificity. PMID- 12883866 TI - Cloning and characterisation of a glucoamylase gene (GlaM) from the dimorphic zygomycete Mucor circinelloides. AB - This article reports a novel strategy for the cloning of glucoamylase genes using conserved sequences and semi-nested PCR and its application in cloning the GlaM glucoamylase gene and cDNA from the dimorphic zygomycete Mucor circinelloides. The deduced 609-amino-acid enzyme (including signal peptide) is 63% identical to the Rhizopus oryzae raw starch-degrading glucoamylase and is the third glucoamylase reported to have the putative starch-binding domain placed N terminally. The C-terminal catalytic domain is separated from the starch-binding domain by a serine/threonine-rich linker. An alignment of the cloned gene and cDNA sequences showed that the gene contains three introns. The transcriptional start site and the site of polyadenylation were defined by primer extension and 3'RACE, respectively. The atypical Kozak sequence is identical to the one used in R. oryzae in positions -1 to -4. Northern slot blots revealed that glucoamylase transcription is induced during growth on starch and repressed by glucose. In silico analysis of the 1.9-kb promoter sequence cloned by inverse PCR revealed the presence of several putative regulatory elements, most notably a 19-bp sequence containing six overlapping copies of the Saccharomyces cerevisiae Nrg1p binding sequence. PMID- 12883867 TI - Development of a biosensor for on-line detection of tributyltin with a recombinant bioluminescent Escherichia coli strain. AB - A biosensor was developed for the detection of tributyltin (TBT), using a bioluminescent recombinant Escherichia coli:: luxAB strain. Dedicated devices allowed the on-line measurement of bioluminescence, pH and dissolved oxygen values and the feed-back regulation of temperature. Bacterial physiology was monitored by the measurement of the cellular density, respiratory activity and the intracellular level of ATP, glucose and acetate levels. Our results showed that a synthetic glucose medium gave a better TBT detection limit than LB medium (respectively 0.02 micro M and 1.5 micro M TBT). High growth and dilution rates ( D=0.9 h(-1)) allowed maximum light emission from the bacterium. Moreover, simple atmospheric air bubbling was sufficient to provide oxygen for growth and the bioluminescence reaction. Real-time monitoring of bioluminescence after TBT induction occurred with continuous addition of decanal up to 300 micro M, which was not toxic throughout a 7-day experiment. The design of our biosensor and the optimization of the main parameters that influence microbial activity led to the capacity for the detection of TBT. PMID- 12883868 TI - Display of a functional hetero-oligomeric catalytic antibody on the yeast cell surface. AB - A functional hetero-oligomeric protein was, for the first time, displayed on the yeast cell surface. A hetero-oligomeric Fab fragment of the catalytic antibody 6D9 can hydrolyze a non-bioactive chloramphenicol monoester derivative to produce chloramphenicol. The gene encoding the light chain of the Fab fragment of 6D9 was expressed with the tandemly-linked C-terminal half of alpha-agglutinin. At the same time, the gene encoding the Fd fragment of the heavy chain of the Fab fragment was expressed as a secretion protein. The combined Fab fragment displayed and associated on the yeast cell surface had an intermolecular disulfide linkage between the light and heavy chains. This protein fragment catalyzed the hydrolysis of a chloramphenicol monoester derivative and exhibited high stability in binding with a transition-state analog (TSA). The catalytic reaction was also inhibited by the TSA. The successful display of a functional hetero-oligomeric catalytic antibody provides a useful model for the display of hetero-oligomeric proteins and enzymes. PMID- 12883869 TI - Antagonistic properties of two recombinant strains of Streptomyces melanosporofaciens obtained by intraspecific protoplast fusion. AB - Intraspecific protoplast fusion was used to produce stable prototrophic recombinants of Streptomyces melanosporofaciens EF-76, a biocontrol agent of plant disease producing geldanamycin. Two recombinant strains (FP-54 and FP-60) that differed with regard to their antagonistic properties against Bacillus cereus ATCC 14579, Streptomyces scabies EF-35 and Phytophthora fragariae var. rubi 390 were characterized. FP-60 lost the ability to inhibit the in vitro growth of these microbial strains while FP-54 exhibited higher antagonistic activities against them. FP-60 was deficient in geldanamycin biosynthesis whereas FP-54 was shown to produce, in addition to geldanamycin, at least two other antimicrobial compounds that were absent in the culture supernatants of strain EF 76. Like the wild-type strain EF-76, strain FP-54 reduced common scab symptoms on potato tuber but no significant difference was observed between the disease index attributed to tubers treated with strain EF-76 or with strain FP-54. Strain FP-60 showed no protective effect against common scab. The disease index of tubers treated with this recombinant was worse than the index associated with potato tubers from control treatments. PMID- 12883870 TI - Effect of cysteine desulfhydrase gene disruption on L-cysteine overproduction in Escherichia coli. AB - In Escherichia coli, the enzyme called cysteine desulfhydrase (CD), which is responsible for L-cysteine degradation, was investigated by native-PAGE and CD activity staining of crude cell extracts. Analyses with gene-disrupted mutants showed that CD activity resulted from two enzymes: tryptophanase (TNase) encoded by tnaA and cystathionine beta-lyase (CBL) encoded by metC. It was also found that TNase synthesis was induced by the presence of L-cysteine. The tnaA and metC mutants transformed with the plasmid containing the gene for feedback-insensitive serine acetyltransferase exhibited higher L-cysteine productivity than the wild type strain carrying the same plasmid. These results indicated that TNase and CBL did act on L-cysteine degradation in E. coli cells. PMID- 12883871 TI - Fermentation characteristics and protein expression patterns in a recombinant Escherichia coli mutant lacking phosphoglucose isomerase for poly(3 hydroxybutyrate) production. AB - For the efficient production of poly(3-hydroxybutyrate) (PHB) using recombinant Escherichia coli, it is of primal importance to overproduce NADPH, which is necessary for the PHB synthetic pathway. In order to overproduce NADPH in the pentose phosphate (PP) pathway, a recombinant E. coli was constructed in which the phosphoglucose isomerase ( pgi) gene was knocked out to force the carbon flow into the PP pathway. The fermentation characteristics of the recombinant E. coli mutant lacking pgi were then investigated to determine the effect of overproduction of NADPH on efficient PHB production. It was found that, compared with the parent strain ( E. coli JM109), growth of the E. coli mutant lacking pgi ( E. coli DF11) is repressed due to NADPH overproduction in the PP pathway. Furthermore, repressed cell growth can be recovered to some extent by introducing a NADPH-consuming pathway, such as the PHB synthetic pathway. Efficient PHB production using such recombinant E. coli (DF11/pAeKG1) could be attained by appropriately controlling the glucose concentration in the fermentor. Total gene expression was investigated at the protein level by two-dimensional electrophoresis. Out of 22 differentially expressed proteins, 12 were identified with the aid of MALDI-TOF mass spectrometry. Variations in the accumulation of PHB in the recombinant pgi mutant carrying phb (E. coli DF11/pAeKG1) corresponded to the expression of proteins encoded by rpsA, znuA, fabD, potD, fkpA, gapA, ynaF and ibpA. The unfavorable conditions generated by PHB accumulation in the pgi mutant carrying phb resulted in the highest expression of 30S ribosomal protein S1, which ultimately caused a further increase in soluble protein synthesis. PMID- 12883872 TI - Roles of tert-butyl formate, tert-butyl alcohol and acetone in the regulation of methyl tert-butyl ether degradation by Mycobacterium austroafricanum IFP 2012. AB - Mycobacterium austroafricanum IFP 2012 is a Gram-positive strain able to grow on methyl tert-butyl ether (MTBE) as a sole carbon and energy source. The effect of two downstream metabolites of MTBE, tert-butyl formate (TBF) and tert-butyl alcohol (TBA) on MTBE degradation was investigated using resting cells. The addition of low concentrations of TBF decreased the MTBE degradation rate by about 30%. In contrast, the addition of TBA did not have a significant effect on MTBE degradation rate, even at high concentrations; and it was also shown that TBA degradation occurred only once MTBE was exhausted. At neutral pH, TBF hydrolysis involved mainly an esterase-type activity regulated by the presence of TBA. The TBF degradation rate was about four times lower than the MTBE degradation rate. Furthermore, acetone was identified as an intermediate during TBA degradation. An acetone mono-oxygenase activity, inhibited by methimazole but not by acetylene, was suggested. It was different from the MTBE/TBA mono oxygenase and, thus, acetone did not appear to compete with MTBE and TBA for the same enzyme. These new results show that the metabolic regulation of the early steps of MTBE degradation by M. austroafricanum IFP 2012 is complex, involving inhibition and competition phenomena. PMID- 12883873 TI - Novel proline hydroxylase activities in the pneumocandin-producing fungus Glarea lozoyensis responsible for the formation of trans 3- and trans 4-hydroxyproline. AB - Novel proline 3-hydroxylase (P3H) and proline 4-hydroxylase (P4H) activities that convert free l-proline to both trans 3- and trans 4-hydroxy- l-proline were detected in protein extracts of the anamorphic fungus Glarea lozoyensis. The enzymatic conversion of l-proline to trans 3- and trans 4-hydroxy- l-proline was strictly dependent on alpha-ketoglutarate, ascorbate, and dithiothreitol. Ferrous iron was required for optimal P3H and P4H activity. These substrate and co-factor requirements indicate these enzyme activities belong to the class of 2 oxoglutarate-dependent dioxygenases. Both P3H and P4H were inhibited by zinc and other trace metals. The addition of proline to the fermentation medium resulted in an increase in the specific activity of P4H and a decrease in the synthesis of pneumocandin C(0). Additionally, the synthesis of trans 3- and trans 4-hydroxy- l proline in vivo was affected differently by the proline concentration in the medium. This result suggested that two enzymes may be responsible for the regio- and stereospecific hydroxylation of l-proline. PMID- 12883874 TI - Quantitative detection of crystalline lysine supplementation in poultry feeds using a rapid bacterial bioluminescence assay. AB - Lysine is an essential amino acid for both humans and animals; and it is usually the first or second limiting amino acid in most formulated diets. In order to estimate the lysine content in feeds and feed sources, rapid amino acid bioassays have been developed. The objective of this work is to assess a rapid assay for lysine supplementation in chicken feeds, using a luminescent Escherichia coli lysine-auxotrophic strain, to avoid prior thermal sterilization. An E. coli lysine auxotroph carrying a plasmid with lux genes was used as the test organism. The lysine assay was conducted using depleted auxotrophic cells in lysine samples. Luminescence was measured with a Dynex MLX luminometer after addition of the aldehyde substrate. Growth response (monitored as optical density at 600 nm) and light emission response of the assay E. coli strain were monitored to generate standard curves. Bioluminescent analysis of feed samples indicated that the method works well in the presence of a complex feed matrix. Comparison of both optical density and luminescent-based methods indicated that, when the assay takes place under optimal conditions, both methodologies correlated well ( r(2)=0.99). Except for the 0.64% lysine-supplemented feed, estimates for lysine based on the bacterial assay were over 80% (82-97%) of the theoretical values. Animal data showed that the bacterial bioluminescent method correlated well with the chick bioassay when diets with different levels of lysine supplementation were assayed for lysine bioavailability ( r(2)=0.97). Luminescent methodology coupled with a bacterial growth assay is a promising technique to assess lysine availability in supplemented animal feeds. PMID- 12883876 TI - Degradation and detoxification of endosulfan isomers by a defined co-culture of two Bacillus strains. AB - The degradation of alpha and beta isomers of endosulfan by a two-member bacterial co-culture was studied. Results were similar whether the two isomers were present individually or together, as in technical endosulfan. The degradation of both isomers was accompanied by the formation of endosulfan diol and endosulfan lactone. Accumulation of the metabolite, endosulfan sulfate was, however, not observed during the reaction with either of the isomers. The microbial degradation of endosulfan isomers was also accompanied by a decrease in its toxicity to the test organism Tubifex tubifex Muller. PMID- 12883875 TI - Removal of inorganic and organic mercurials by immobilized bacteria having mer ppk fusion plasmids. AB - Feasibility of biological mercury removal from wastewater was examined by using alginate-immobilized cells of Escherichia coli carrying mer-ppk fusion plasmid pMKB18. Immobilized cells engineered to express mercury-transport system, organomercurial lyase and polyphosphate efficiently removed organic and inorganic mercury from contaminated wastewater over a wide concentration range of mercurials, probably via intracellular accumulation mediated by ppk-specified polyphosphate. Bioaccumulation of mercury was selective compared to other metals such as Cd(2+), Pb(2+) and Cr(6+). The immobilized cells could be used repeatedly (at least three times) without large loss of mercury removal activity. From these results, it is concluded that the mer-ppk fusion plasmid and the immobilized cells are useful for simultaneous removal of organic and inorganic mercury from contaminated wastewater. PMID- 12883877 TI - A novel catabolic activity of Pseudomonas veronii in biotransformation of pentachlorophenol. AB - Pseudomonas veronii PH-05, a bacterial strain capable of transforming pentachlorophenol (PCP) to a metabolic intermediate, was isolated by selective enrichment of soil samples from a timber storage yard. Strain PH-05 was shown to be able to grow using PCP as the sole source of carbon and energy. GC-MS analysis showed that the metabolic intermediate was tetrachlorocatechol, which inhibited the growth of this strain. The formation of tetrachlorocatechol during biotransformation was monitored, and its inhibitory effect on growth of strain PH 05 was analyzed at a range of concentrations. The catabolic activity of the isolated strain differs from that of other PCP-degrading bacteria, which metabolize PCP through a chlorinated hydroquinone intermediate. PMID- 12883878 TI - Prostate lymphoscintigraphy for sentinel lymph node identification in canines: reproducibility, uptake, and biokinetics depending on different injection strategies. AB - At present there are neither clinical nor experimental data available on the influence of technical details on the quality and reproducibility of prostate lymphoscintigraphy. Six adult fox hounds received repeated transrectal ultrasound guided intraprostatic injections of a technetium 99m labeled nanocolloid to prove the influence of different techniques of injection (one central injection in both prostate lobes vs two peripheral injections in both lobes) on tracer accumulation in sentinel lymph nodes (SLN) and other organs. The reproducibility of the favored technique was examined and in a last step it was subject to scrutiny following a reduction of the injected volume to 1% of the prostate volume. The number of scintigraphically visualized SLN varied between four and seven. They were located in the region of the internal and external iliac vessels, presacrally, paravesically, and directly paraprostatically. In five of six cases, the localization was reproducible both with the central application of an identical volume as well as with the volume reduced central injection. Tracer accumulation of SLNs and other organs varied enormously. We expect that with the combination of both injection techniques, even with the reduced injection volume, an optimized prostate lymphoscintigraphy will be the outcome. PMID- 12883879 TI - Loss of heterozygosity and methylation of p16 in renal cell carcinoma. AB - To investigate the possible role of genomic aberrations of chromosome 9p21 in the tumorigenesis of human renal cell carcinoma (RCC), 40 sporadic RCCs were studied using PCR analyses. The tumours were predominantly low stage and low grade. Loss of heterozygosity (LOH) was observed in nine of 39 informative cases, but no homozygous deletion was noticed. Hypermethylation of the promoter region of p16 occurred in eight of the 40 RCCs. No correlation was found between hypermethylation of the p16 gene and LOH on 9p21. A similar level of LOH and methylation was observed in the 40 RCCS regardless of histology, grade and stage. These results suggest that inactivation of p16 and the possibility of other unknown tumour suppressor genes located on other chromosomes could be involved in the pathogenesis of RCC. PMID- 12883880 TI - Expression of functional protease-activated receptor 1 in human prostate cancer cell lines. AB - Functional protease-activated receptors (PAR) are expressed by a variety of malignant cells. In the present study, RT-PCR assays demonstrated the expression of the thrombin receptor PAR-1 mRNA in human prostate cancer cell lines DU 145, LnCAP, and SV40-immortalized human prostate epithelial cell line PNT1A. In contrast, the additional thrombin receptors PAR-3 and PAR-4 were not detected. PAR-1 protein localized to the cellular surface was detected by flow cytometry in all three cell lines. To demonstrate the functional importance of the PAR-1, the effects of different concentrations of thrombin on cell proliferation kinetics were assessed. The treatment of growth-arrested cells with varying concentrations of thrombin demonstrated dose- and time-dependent effects. At low concentration (<0.5 U/ml), thrombin induced proliferation of all prostate-derived cell lines. Thrombin at higher concentration (1.0 U/ml) initially stimulated PNT1A and LnCAP cells to proliferate (time of thrombin application 24 h and 48 h) followed by inhibited growth when assessed after 72 h of incubation. In contrast, 1.0 U/ml thrombin caused earlier inhibition of DU 145 proliferation starting at 48 h of incubation. Our results suggest that PAR-1 mediates the proliferation-modulating effects of thrombin on prostate cancer cells. PMID- 12883881 TI - Cell cycle events during the development of the silk glands in the mulberry silkworm Bombyx mori. AB - Silk glands of the mulberry silkworm Bombyx mori are long and paired structures originating from the labial region and are anatomically and physiologically divided into three major compartments, the anterior, middle and posterior silk glands. The silk gland morphogenesis is complete by 8 days post egg laying. Extensive growth of silk glands during the larval stages is due to increase in tissue mass and not cell number. The cells in a completely formed silk gland pursue an endoreplicative cell cycle, and the genome undergoes multiple rounds of replication without mitosis or nuclear division. The expression patterns of cyclin B (mitotic cyclin) and cyclin E (G1 cyclin, essential for G1/S transition in both mitotic and endoreplicative cell cycles) in the course of silk gland development revealed that mitotic cell divisions take place only in the apex of the growing silk gland. However, the persistence of another mitotic focus in the middle silk gland even when the growing apex has moved well past this zone suggested the continued operation of mitosis for a while in this restricted region. The lack of cyclin B expression and abundance of cyclin E in the rest of the areas confirmed an alternation of the G1 and S phases of the cell cycle without an intervening mitotic phase. No expression of cyclin B was noticed anywhere in the silk glands after stage 25 of embryogenesis, indicating a complete switch over to the endomitotic mode of the cell cycle. The onset of expression of various genes encoding different silk proteins correlated with the onset of endomitotic events. PMID- 12883882 TI - Expression of developmental genes during early embryogenesis of Hydra. AB - Hydra is a classical model to study key features of embryogenesis such as axial patterning and stem cell differentiation. In contrast to other organisms where these mechanisms are active only during embryonic development, in Hydra they can be studied in adults. The underlying assumption is that the machinery governing adult patterning mimics regulatory mechanisms which are also active during early embryogenesis. Whether, however, Hydra embryogenesis is governed by the same mechanisms which are controlling adult patterning, remains to be shown. In this paper, in precisely staged Hydra embryos, we examined the expression pattern of 15 regulatory genes shown previously to play a role in adult patterning and cell differentiation. RT-PCR revealed that most of the genes examined were expressed in rather late embryonic stages. In situ hybridization, nuclear run-on experiments, and staining of nucleolar organizer region-associated proteins indicated that genes expressed in early embryos are transcribed in the engulfed "nurse cells" (endocytes). This is the first direct evidence that endocytes in Hydra not only provide nutrients to the developing oocyte but also produce maternal factors critical for embryogenesis. Our findings are an initial step towards understanding the molecular machinery controlling embryogenesis of a key group of basal metazoans and raise the possibility that in Hydra there are differences in the mechanisms controlling embryogenesis and adult patterning. PMID- 12883883 TI - Characterization of a novel cellulose synthesis inhibitor. AB - The physiological effects of an experimental herbicide and cellulose synthesis inhibitor, N2-(1-ethyl-3-phenylpropyl)-6-(1-fluoro-1-methylethyl)-1,3,5-triazine 2,4-diamine, called AE F150944, are described. In the aminotriazine molecular class, AE F150944 is structurally distinct from other known cellulose synthesis inhibitors. It specifically inhibits crystalline cellulose synthesis in plants without affecting other processes that were tested. The effects of AE F150944 on dicotyledonous plants were tested on cultured mesophyll cells of Zinnia elegans L. cv. Envy, which can be selectively induced to expand via primary wall synthesis or to differentiate into tracheary elements via secondary wall synthesis. The IC50 values during primary and secondary wall synthesis in Z. elegans were 3.91 x 10(-8) M and 3.67 x 10(-9) M, respectively. The IC50 in suspension cultures of the monocot Sorghum halapense (L.) Pers., which were dividing and synthesizing primary walls, was 1.67 x 10(-10) M. At maximally inhibitory concentrations, 18-33% residual crystalline cellulose synthesis activity remained, with the most residual activity observed during primary wall synthesis in Z. elegans. Addition to Z. elegans cells of two other cellulose synthesis inhibitors, 1 microM 2,6-dichlorobenzonitrile and isoxaben, along with AE F150944 did not eliminate the residual cellulose synthesis, indicating little synergy between the three inhibitors. In differentiating tracheary elements, AE F150944 inhibited the deposition of detectable cellulose into patterned secondary wall thickenings, which was correlated with delocalization of lignin as described previously for 2, 6-dichlorobenzonitrile. Freeze-fracture electron microscopy showed that the plasma membrane below the patterned thickenings of AE F150944 treated tracheary elements was depleted of cellulose-synthase-containing rosettes, which appeared to be inserted intact into the plasma membrane followed by their rapid disaggregation. AE F150944 also inhibited cellulose-dependent growth in the rosette-containing alga, Spirogyra pratensis, but it did not inhibit cellulose synthesis in Acetobacter xylinum or Dictyostelium discoideum, both of which synthesize cellulose via linear terminal complexes. Therefore, AE F150944 may inhibit crystalline cellulose synthesis by destabilizing plasma membrane rosettes. PMID- 12883884 TI - Intron-mediated enhancement of gene expression in transgenic plants using chimeric constructs composed of the Peanut chlorotic streak virus (PClSV) promoter-leader and the antisense orientation of PClSV ORF VII (p7R). AB - The antisense orientation of the Peanut chlorotic streak virus (PClSV) open reading frame (ORF) VII (denoted as p7R), in conjunction with the sense orientation of the PClSV leader sequence, acts as an intron and enhances the expression of a reporter gene, analyzed in protoplasts and transgenic plants of tobacco ( Nicotiana tabacum L.). Correct 5' and 3' splicing sites were determined for intron removal from the chimeric constructs using either beta-glucuronidase (GUS) or chloramphenicol acetyltransferase (CAT) as a reporter gene. In this splicing process, the active consensus 5' splicing donor site (AG/GTATA) is located at position +283 to +289 from the transcription start site (TSS) of the PClSV full-length transcript (FLt). The 3' splice site (TAG/GATT) is located on the p7R sequence at position +785 to +791 from the TSS. The combination of PClSV FLt leader and p7R enhanced the expression of reporter genes (CAT and GUS) by as much as 2-fold compared to the strong constitutive PClSV FLt promoter without an interfering leader sequence and about 30- to 800-fold compared to constructs containing the sense orientation of PClSV ORF VII (p7) in both protoplast transient-expression experiments and stably transformed transgenic plants. An increased level of mature transcripts accompanied this. This suggests that this combination of elements can mediate the intron-mediated enhancement (IME) phenomenon. We also demonstrated comparative IME with other heterologous promoters from caulimoviruses. PMID- 12883885 TI - Light-induced fluorescence changes in Phycomyces: evidence for blue light receptor associated flavo-semiquinones. AB - Light-induced fluorescence changes (LIFCs) were detected in sporangiophores of the blue-light-sensitive fungus Phycomyces blakesleeanus (Burgeff). The LIFCs can be utilized as a spectrophotometric assay for blue-light photoreceptors and for the in vivo characterization of their photochemical primary reactions. Blue-light irradiation of sporangiophores elicited a transient decrease and subsequent regeneration of flavin-like fluorescence emission at 525 nm. The signals recovered in darkness in about 120 min. In contrast to blue light, near-UV (370 nm) caused an increase in the fluorescence emission at 525 nm. Because the LIFCs were altered in a light-insensitive madC mutant with a defective photoreceptor, the fluorescence changes must be associated with early photochemical events of the transduction chain. Action spectra for the fluorescence changes at 525 nm showed major peaks near 470 and 600 nm. Double-pulse experiments involving two consecutive pulses of either blue and near-UV, blue and red, or near-UV and red showed that the responses depended on the sequence in which the different wavelengths were applied. The results indicate a blue-light receptor with intermediates in the near-UV, blue and red spectral regions. We explain the results in the framework of a general model, in which the three redox states of the flavin photoreceptor, the oxidized flavin (Fl), the flavo-semiquinone (FlH*), and the flavo-hydroquinone (FlH2) are each acting as chromophores with their own characteristic photochemical primary reactions. These consist of the photoreduction of the oxidized flavin generating semiquinone, the photoreduction of the semiquinone generating hydroquinone, and the photooxidation of the flavo hydroquinone regenerating the pool of oxidized flavins. The proposed mechanism represents a photocycle in which two antagonistic photoreceptor forms, Fl and FlH2, determine the pool size of the biological effector molecule, the flavo semiquinone. The redox changes that are associated with the photocycle are maintained by redox partners, pterins, that function in the near-UV as secondary chromophores. PMID- 12883886 TI - Anaerobic induction of the maize GapC4 promoter in poplar leaves requires light and high CO2. AB - The maize (Zea mays L.) glyceraldehyde-3-phosphate dehydrogenase gene 4 ( GapC4) promoter confers anaerobic gene expression in tobacco (Nicotiana tabacum L.), potato (Solanum tuberosum L.) and Arabidopsis thaliana (L.) Heynh. Here we have investigated its expression in hybrid poplar (Populus tremula x P. alba). Our results show that the promoter is not expressed in leaves and stems under normoxic conditions while anaerobiosis induces reporter gene expression in leaves up to a level observed for the STLS-1 promoter from potato that is shown to confer leaf-specific gene expression in transgenic poplar. Anaerobic induction is cell autonomous and requires a CO2 atmosphere and light. As in tobacco, the GapC4 promoter in poplar is wound inducible. The induction by CO2 and light may reflect a natural situation because flooding, a natural cause of anaerobiosis, is often accompanied by high CO2 concentrations in the floodwater. Our results show that the GapC4 promoter is suitable as an anaerobic reporter and as an inducible gene expression system in poplar. PMID- 12883887 TI - Slippery surfaces of carnivorous plants: composition of epicuticular wax crystals in Nepenthes alata Blanco pitchers. AB - Plants in the genus Nepenthes obtain a substantial nutrient supply by trapping insects in highly modified leaves. A broad zone of the inner surface of these pitchers is densely covered with wax crystals on which most insects lose their footing. This slippery wax surface, capturing prey and preventing its escape from the trap, plays a pivotal role in the carnivorous syndrome. To understand the mechanism of slipperiness, the present investigation aimed at an ultrastructural and physico-chemical characterization of the wax crystals in pitchers of N. alata Blanco. Scanning electron microscopy revealed that entire platelets protruded perpendicularly from the surface. Methods were developed that allowed the mechanical removal of wax crystals from the pitcher surface. It could be shown that the sampling was selective for the epicuticular wax, relevant for plant insect interactions. The crystals consisted of a mixture of aliphatic compounds dominated by very-long-chain aldehydes. Triacontanal, at 43% the most abundant constituent, was largely responsible for crystal formation. Solubility data indicate that the Nepenthes crystals contained polymeric forms of this aldehyde. The resulting mechanical properties of the polymer crystals and the mechanism of slipperiness are discussed. PMID- 12883888 TI - SulP, a nuclear gene encoding a putative chloroplast-targeted sulfate permease in Chlamydomonas reinhardtii. AB - Genomic, proteomic, phylogenetic and evolutionary aspects of a novel gene encoding a putative chloroplast-targeted sulfate permease of prokaryotic origin in the green alga Chlamydomonas reinhardtii are described. This nuclear-encoded sulfate permease gene (SulP) contains four introns, whereas all other known chloroplast sulfate permease genes lack introns and are encoded by the chloroplast genome. The deduced amino acid sequence of the protein showed an extended N-terminus, which includes a putative chloroplast transit peptide. The mature protein contains seven transmembrane domains and two large hydrophilic loops. This novel prokaryotic-origin gene probably migrated from the chloroplast to the nuclear genome during evolution of C. reinhardtii. The SulP gene, or any of its homologues, has not been retained in vascular plants, e.g. Arabidopsis thaliana, although it is encountered in the chloroplast genome of a liverwort (Marchantia polymorpha). A comparative structural analysis and phylogenetic origin of chloroplast sulfate permeases in a variety of species is presented. PMID- 12883889 TI - Down-regulation of linear and activation of cyclic electron transport during drought. AB - The effects of short-term drought on the regulation of electron transport through photosystems I and II (PSI and PSII) have been studied in Hordeum vulgare L. cv. Chariot. Fluorescence measurements demonstrated that electron flow through PSII decreased in response to both drought and CO2 limitation. This was due to regulation, as opposed to photoinhibition. We demonstrate that this regulation occurs between the two photosystems--in contrast to PSII, PSI became more oxidised and the rate constant for P700 re-reduction decreased under these conditions. Thus, when carbon fixation is inhibited, electron transport is down regulated to match the reduced requirement for electrons and minimise reactive oxygen production. At the same time non-photochemical quenching (NPQ) increases, alleviating the excitation pressure placed on PSII. We observe an increase in the proportion of PSI centres that are 'active' (i.e. can be oxidised with a saturating flash and then rapidly re-reduced) under the conditions when NPQ is increased. We suggest that these additional centres are primarily involved in cyclic electron transport, which generates the DeltapH to support NPQ and protect PSII. PMID- 12883890 TI - Changes in growth conditions alter the male strobilus gene expression pattern in Cryptomeria japonica. AB - Two-year old saplings grown from cuttings of Cryptomeria japonica D. Don initiate strobilus development following treatment with gibberellic acid under long-day photoperiods. At 25 degrees C with a 14-h photoperiod in a phytotron, male strobili initiated normally; however, they remained green and fell from the saplings prematurely. To examine the change in male strobilus development at the molecular level, three genes expressed specifically in male strobili were analyzed. Two were MADS box genes homologous to the B-function genes in angiosperms, CjMADS1 and CjMADS2, and the third was Cry j I, which encodes an allergen protein, and this gene is expressed mainly in microspores. Under phytotron growing conditions, the homeotic genes were expressed constantly, which reflected the extended early developmental stage of male strobili. On the other hand, Cry j I expression was detected after a long delay just before strobilus development ceased. These results indicate that the expression of the genes related to male reproductive development in C. japonica is regulated by a factor(s) that is sensitive to environmental signals. PMID- 12883891 TI - The SLC22 drug transporter family. AB - The SLC22 family comprises organic cation transporters (OCTs), zwitterion/cation transporters (OCTNs), and organic anion transporters (OATs). These transporters contain 12 predicted alpha-helical transmembrane domains (TMDs) and one large extracellular loop between TMDs 1 and 2. Transporters of the SLC22 family function in different ways: (1) as uniporters that mediate facilitated diffusion in either direction (OCTs), (2) as anion exchangers (OAT1, OAT3 and URAT1), and (3) as Na(+)/ l-carnitine cotransporter (OCTN2). They participate in the absorption and/or excretion of drugs, xenobiotics, and endogenous compounds in intestine, liver and/or kidney, and perform homeostatic functions in brain and heart. The endogenous substrates include monoamine neurotransmitters, choline, l carnitine, alpha-ketoglutarate, cAMP, cGMP, prostaglandins, and urate. Defect mutations of transporters of the SLC22 family may cause specific diseases such as "primary systemic carnitine deficiency" or "idiopathic renal hypouricemia" or change drug absorption or excretion. PMID- 12883892 TI - Cariporide enhances lactate clearance upon reperfusion but does not alter lactate accumulation during global ischaemia. AB - Cariporide (HOE 642) inhibits the Na+/H+ exchanger and would be expected to reduce lactate accumulation during ischaemia and stimulate lactate/H+ co transporter upon reperfusion. The aim of this study was to determine the effect of cariporide on lactate production during global ischaemia and release during reperfusion. Guinea-pig hearts perfused in the Langendorff mode were exposed to 45 min global ischaemia and 30 min reperfusion with or without cariporide (5 or 10 micromol/l). Cardiac function was assessed by measurement of left ventricular developed pressure (LVDP). Lactate and pH were measured in coronary effluent before ischaemia and throughout reperfusion. Tissue metabolites (lactate, adenine nucleotides, guanine nucleotides and purine) were measured in ventricular biopsy samples collected at the beginning and end of ischaemia. Cariporide significantly improved recovery of LVDP (from 66% for control to 88% and 93% for 5 and 10 micromol/l cariporide, respectively). During ischaemia, only 10 micromol/l cariporide produced a small (10%) but significant preservation of ATP and GTP compared to control. This was associated with significant reduction (25%) in ischaemic contracture. Cariporide did not influence lactate accumulation during ischaemia but significantly increased lactate efflux (18%) during the first 60 s of reperfusion. In conclusion, cariporide does not alter lactate accumulation during ischaemia but enhances lactate efflux upon reperfusion, which may have implications for its cardioprotective action. PMID- 12883893 TI - "In vivo" monitoring of neuronal network activity in zebrafish by two-photon Ca(2+) imaging. AB - The zebrafish larva is a powerful model for the analysis of behaviour and the underlying neuronal network activity during early stages of development. Here we employ a new approach of "in vivo" Ca(2+) imaging in this preparation. We demonstrate that bolus injection of membrane-permeable Ca(2+) indicator dyes into the spinal cord of zebrafish larvae results in rapid staining of essentially the entire spinal cord. Using two-photon imaging, we could monitor Ca(2+) signals simultaneously from a large population of spinal neurons with single-cell resolution. To test the method, Ca(2+) transients were produced by iontophoretic application of glutamate and, as observed for the first time in a living preparation, of GABA or glycine. Glycine-evoked Ca(2+) transients were blocked by the application of strychnine. Sensory stimuli that trigger escape reflexes in mobile zebrafish evoked Ca(2+) transients in distinct neurons of the spinal network. Moreover, long-term recordings revealed spontaneous Ca(2+) transients in individual spinal neurons. Frequently, this activity occurred synchronously among many neurons in the network. In conclusion, the new approach permits a reliable analysis with single-cell resolution of the functional organisation of developing neuronal networks. PMID- 12883894 TI - A comparative assessment of two techniques for investigating initial cardiovascular reflexes under acute orthostatic stress. AB - The physiological differences between active and passive changes in posture have been previously established. This study determined the extent of the differences in the initial cardiovascular responses to the passive head-up tilt (HUT) and the active squat-stand test (SST). Eleven females and 13 males underwent one +75 degrees HUT and one SST. Beat-to-beat diastolic blood pressure (DBP), systolic blood pressure (SBP), mean arterial pressure (MAP) and heart rate (HR) were determined non-invasively. Data were recorded 10 s prior to (control) and 30 s after tilt or stand (event). Blood pressure and HR responses were analysed by calculating the deviation from control at 10 s (T10), 20 s (T20) and 30 s (T30) after the onset of each test. The DBP response (reflecting changes in systemic vascular resistance) at T10 was -10 (2) mmHg [mean (SEM)] for the HUT and -25 (2) mmHg for the SST ( P<0.01). DBP returned to control levels by T30 for the HUT, but remained depressed for the SST. MAP responses directly reflected these changes in DBP. HR significantly increased from control values ( P<0.001) for the HUT [+14 (1) bpm] and the SST [+16 (1) bpm], and remained elevated for the entire 30-s period for both tests. This study demonstrates that although the magnitude of the initial blood pressure decrease is greater for the active SST compared with the passive HUT, the reflex compensatory response is no different, making the SST a greater challenge for the cardiovascular reflexes. PMID- 12883895 TI - Effects of long- and short-term fatiguing stretch-shortening cycle exercises on reflex EMG and force of the tendon-muscle complex. AB - This study examined the fatigue effects of stretch-shortening cycle exercises of different intensity and duration on stretch reflex EMG and mechanical responses of the triceps surae muscle. Twelve subjects performed either a 10-km run ( n=6) or short but exhaustive rebound exercise on a sledge apparatus ( n=6). Passive reflex tests (mechanically induced ankle dorsiflexions) were examined before, after as well as 2 h, 2 and 7 days after exercise. Mechanical reflex responses were recorded from the ergometer torque signal. An acute contractile failure was observed as large reductions in twitch responses, especially in the sledge subgroup who showed high post-exercise peak blood lactate and an increased EMG/torque ratio. Independently of the exercise, the delayed fatigue analysis revealed strong relationships between the reflex-induced EMG and mechanical changes. In addition to muscle damage, these results may be explained by inhibitory effects via the sensitisation of small muscle afferents particularly during the exercise-induced delayed recovery process. PMID- 12883896 TI - Comparison between young and older women in explosive power output and its determinants during a single leg-press action after optimisation of load. AB - Lower limb explosive power, which is more predictive of functional difficulties than strength per se with women being more at risk than men for disability, has been previously compared between young and older women using systems with fixed inertia. Individuals may have been obliged to use a percentage of their maximum strength that is not ideal for performing the movement at the optimum speed for maximum power output. This study was designed to compare explosive power output and its two determinants, optimal force and optimal speed, during a leg-press action between young and older women after optimising the load for maximum power production. The experiments were carried out on 20 women in good physical condition: 10 older, aged between 65 and 74 years and 10 young, aged between 18 and 30. Explosive power output was measured by setting the initial load at different percentages of maximum isometric strength and measuring the corresponding speed of movement during a leg-press action of the dominant leg. Maximum peak power, which was obtained at 60% of maximum isometric strength in both young and older women, was 61% lower in the older women ( P<0.0001). This was due to a 52% lower optimal force ( P<0.0001) and 21% lower optimal speed ( P<0.01). The ratio of peak power to maximum isometric strength was 22.1% lower in the older women ( P<0.01). After optimising the load, both lower speed of movement and lower strength determine the lower levels of power in older women. Power is more affected by ageing than isometric strength. PMID- 12883897 TI - Effect of menstrual cycle and gender on ventilatory and heart rate responses at the onset of exercise. AB - To clarify the luteal-follicular and male-female differences in ventilatory and heart rate responses at the onset of exercise, seven women and seven men performed voluntary exercise and passive movement for 20 s (brief voluntary exercise and brief passive movement) and voluntary exercise for 3 min (long voluntary exercise) in a sitting position. Voluntary exercise consisted of alternate flexion-extensions of both lower legs with a weight corresponding to about 2.5% of the subjects' body mass attached to each ankle, at a frequency of about 60 times min(-1). Passive movement was carried out without weights by experimenters pulling ropes attached to both of the subjects' ankles, in the same way as voluntary exercise. During these exercises and movements, minute inspiratory ventilation ( V(I)) and heart rate (HR) were continuously measured by breath-by-breath and beat-to-beat techniques. We calculated relative changes of V(I) and HR (Delta V(I) and DeltaHR). Additionally, we averaged Delta V(I) and DeltaHR obtained during the exercise and movement for each subject, and performed a correlation analysis between the averaged Delta V(I) and DeltaHR. It was clarified that: (1) Delta V(I) and DeltaHR in the follicular phase were almost equal to those in the luteal phase; (2) there were no significant male-female differences in these parameters; (3) significant positive correlations were found in both genders only during brief voluntary exercise. We conclude that ventilatory and HR responses at the onset of voluntary exercise and passive movement are not affected by the menstrual cycle or gender. PMID- 12883898 TI - The VO2 response to exhaustive square wave exercise: influence of exercise intensity and mode. AB - We investigated the oxygen uptake ( VO(2)) response to exhaustive square wave exercise of approximately 2, 5 and 8 min duration in cycling and running. Nine males completed a ramp test and three square wave tests on a motorised treadmill and the same four tests on a cycle ergometer, throughout which gas exchange was assessed (Douglas bag method). The peak VO(2) from the ramp test was higher for running than for cycling [mean (SD): 58.4 (2.8) vs. 55.9 (3.7) ml.kg(-1).min(-1); P=0.04]. However VO(2max) (defined as the highest VO(2) achieved in any of the four tests) did not differ between running and cycling [60.0 (2.9) vs. 58.5 (3.3) ml.kg(-1).min(-1); P=0.15]. The peak VO(2) was similar ( P>0.1) for the 5 and 8 min square wave tests [98.5 (1.8) and 99.2 (2.3) % VO(2max) for running; 97.0 (4.2) and 97.5 (2.0) % VO(2max) for cycling] but lower ( P<0.001) for the 2-min test [91.8 (2.5) and 89.9 (5.5) % VO(2max) for running and cycling respectively]. VO(2) increased over the final two 30-s collection periods of the 2-min test for cycling [Delta VO(2)=0.18 (0.15) l.min(-1); P<0.01] but not running [Delta VO(2)=0.00 (0.09) l.min(-1); P=0.98]. We conclude that in the aerobically fit the peak VO(2) for square wave running or cycling at an intensity severe enough to result in exhaustion in approximately 2 min is below VO(2max). In running, VO(2) plateaus at this sub-maximal rate. PMID- 12883899 TI - Influence of pedalling rate on the energy cost of cycling in humans. PMID- 12883900 TI - Comments on "Influence of pedalling rate on the energy cost of cycling in humans". PMID- 12883902 TI - Exercise starts and ends in the brain. AB - Classically the limit to endurance of exercise is explained in terms of metabolic capacity. Cardio-respiratory capacity and muscle fatigue are thought to set the limit and the majority of studies on factors limiting endurance exercise discuss issues such as maximal oxygen uptake (VO2max), aerobic enzyme capacity, cardiac output, glycogen stores, etc. However, this paradigm does not explain the limitation to endurance exercise with large muscle groups at altitude, when at exhaustion exercise is ended without limb locomotor muscle fatigue and with sub maximal cardiac output. A simple fact provides a basis for an explanation. Voluntary exercise starts and ends in the brain. It starts with spatial and temporal recruitment of motor units and ends with their de-recruitment. A conscious decision precedes a voluntary effort. The end of effort is again volitional and a forced conscious decision to stop precedes it, but it is unknown what forces the off-switch of recruitment at exhaustion although sensation of exertion certainly plays a role. An alternative model explaining the limitation of exercise endurance thus proposes that the central nervous system integrates input from various sources all related to the exercise and limits the intensity and duration of recruitment of limb skeletal muscle to prevent jeopardizing the integrity of the organism. This model acknowledges the cardio-respiratory and muscle metabolic capacities as prime actors on the performance scene, while crediting the central nervous system for its pivotal role as the ultimate site where exercise starts and ends. PMID- 12883903 TI - Blood flow in the tibialis anterior muscle by photoplethysmography during foot transmitted vibration. AB - Photoplethysmography (PPG) was used to detect changes in the anterior tibial muscle blood flow (MBF) during foot-transmitted vibration in six healthy subjects. A filter was developed for reducing the vibration-induced artefacts in the PPG signal. The vibration applied was random with constant acceleration power density in the range 5-2000 Hz with an acceleration of 16-46 m s(-2) (rms). The application of the filter technique showed a dramatic reduction in the vibration induced artefacts and distortion of the PPG signal was negligible. The mean ratio and correlation coefficient of MBF originating from filtered and non-filtered PPG signals was [mean (SD)] 0.89 (0.04) and 0.99, respectively. This made it possible to detect a relative increase in MBF of 20% during an acute vibration exposure. The results suggest that the use of the filter enables MBF to be measured by PPG during vibration, thus extending the range of applications of the PPG technique to include ergonomic conditions. PMID- 12883904 TI - Blood coagulation and fibrinolysis after long-duration treadmill exercise controlled by individual anaerobic threshold. AB - For rehabilitation training it is recommended that the intensity of exercise should be clearly below the individual anaerobic threshold (IAT). We investigated blood coagulation, particularly endogenous thrombin potential (ETP) and fibrinolysis following a standardized treadmill (TR) ergometer test at 90% IAT for 60-120 min. Sixteen healthy male non-smokers underwent the TR test. Blood samples were taken after a 30-min rest, immediately after exercise, and 2 h after exercise completion. Extrinsic and intrinsic total (TTP(ex+in)) and endogenous (ETP(ex+in)) thrombin potential, prothrombin fragment 1+2 (F1+2), thrombin antithrombin complex (TAT), plasmin-alpha2-antiplasmin complex (PAP), D-dimer, tissue plasminogen activator antigen and activity (tPA-AG and tPA-ACT) and plasminogen activator inhibitor type 1 antigen and activity (PAI-1-AG and PAI-1 ACT) were measured. Immediately after TR, F1+2, TAT and TTP(ex+in) were increased ( P<0.05) while ETP(ex+in) remained unchanged. In contrast, PAP, D-dimer, tPA-AG, tPA-ACT ( P<0.05) were distinctly enhanced while PAI-1-ACT was decreased ( P<0.05) immediately after exercise. The changes in tPA-AG, tPA-ACT, and PAI-1-ACT were reversed to nearly baseline while the enhancement in PAP and D-dimer was prolonged by more than 2 h after exercise. Long-duration exercise between 60 and 120 min controlled by IAT (90%) on a TR ergometer only implicates a small increase in thrombin generation markers and total (free and alpha(2) macroglubulin-bound thrombin), but not in endogenous (free) thrombin potential alone. In contrast, fibrinolysis is distinctly increased after this type of exercise. Endurance exercise with an intensity below 90% IAT and a duration below 2 h generates a more favourable condition for fibrinolysis than for blood coagulation in healthy young subjects. Data are given as mean (SD). PMID- 12883905 TI - Expression patterns of cartilage collagens and Sox9 during mouse heart development. AB - A majority of congenital heart defects are due to abnormal development of the valves and membranous septa, i.e., connective tissue components of the heart. During development, an interesting feature of cardiac connective tissue is transient expression of collagens typical for cartilage. To better understand the role of these collagens in the heart, we have performed a systematic study on the temporospatial expression of type II and IX collagen isoforms during mouse heart development employing northern hybridization and RNase protection assay. The mRNAs for alpha1(II) and alpha1(IX) collagens were expressed transiently between embryonic days 10.5 and 14.5 in embryonic mouse heart. RNase protection assays revealed that for both transcripts the embryonic ("prechondrogenic") variants of the alternatively spliced mRNA isoforms dominated. Immunohistochemistry demonstrated that type IIA collagen and Sox9, its key transcriptional regulator, were expressed in the epithelial-mesenchymal areas of the developing heart, with partially overlapping patterns particularly in valvular and septal regions. In addition, Sox9 expression was detected widely in the developing heart. These observations support the hypothesis that cartilage collagens, especially the long isoform of type II collagen, participate in the morphogenesis of cardiac valves and septa. PMID- 12883906 TI - Dissociation of mammalian Polycomb-group proteins, Ring1B and Rae28/Ph1, from the chromatin correlates with configuration changes of the chromatin in mitotic and meiotic prophase. AB - The Polycomb group (PcG) gene products form complexes that regulate chromatin configuration to mediate cellular memory to postmitotic somatic cells and postmeiotic oocytes in Drosophila melanogaster. Structural and functional similarities of PcG proteins between invertebrates and vertebrates suggest mammalian PcG proteins may be involved to imprint transcriptional status at various loci into postmitotic and postmeiotic daughter cells. To address molecular mechanisms underlying PcG-mediated cellular memory, it might be a prerequisite to understand subcellular localization of PcG proteins during mitosis and meiosis. In this study, we analyzed subcellular localization of Rae28/Ph1 and Ring1B by using newly generated monoclonal antibodies in mitotic somatic cells and meiotic mouse oocytes. Results suggest that Rae28/Ph1 and Ring1B dissociate from the chromatin upon its condensation in mitotic prophase in the U2-OS human osteosarcoma cell line. During maturation of oocytes, significant alterations of Rae28/Ph1 and Ring1B localization are concordant with configuration changes of the chromatin at the germinal vesicle stage of meiotic prophase. Importantly, dissociation of Rae28/Ph1 and Ring1B from the chromatin temporally correlates with transcriptional arrest both in mitosis and meiosis. Present and previous observations suggest molecular mechanisms required for mitotic regulation of RNA polymerase II could be involved in dissociation of PcG proteins. PMID- 12883907 TI - Brefeldin A-regulated retrograde transport into the endoplasmic reticulum of internalised wheat germ agglutinin. AB - The effects of the fungal metabolite brefeldin A (BFA) on the endocytic routes of internalised wheat germ agglutinin (WGA) were studied in human HepG2 hepatoma cells, drawing particular attention to the application times in relation to the membrane dynamics occurring at the trans Golgi face during endocytosis. As shown in previous studies, transport of internalised WGA into the Golgi apparatus can be classified in three stages being characterised by predominance of vesicular endosomes (stage I), formation of an extended endocytic trans Golgi network (stage II) and uptake of WGA into the stacked Golgi cisternae (stage III). BFA treatment of the cells led to rapid tubular-reticular transformations of the Golgi stacks. Retrograde transport and further destinations of internalised WGA depended on the time of BFA application. When BFA was administered during stages I or II, WGA was localised within the BFA-induced tubules and networks, but never was found within the endoplasmic reticulum. By contrast, BFA treatment during stage III led to a redistribution of internalised WGA into cisternae of the endoplasmic reticulum. These results show that BFA administered according to a precise time schedule can be used as a regulatory agent that allows to control retrograde traffic of internalised molecules into the endoplasmic reticulum. PMID- 12883908 TI - Changes in the choroidal circulation of rabbit following RPE removal. AB - PURPOSE: To examine the effects of surgical removal of the retinal pigment epithelium (RPE) on the choroidal circulation of the rabbit. METHODS: The retina and choroid were examined by biomicroscopy, scanning laser ophthalmoscopy, fluorescein and indocyanine green angiography and histology at various times after the surgical removal of the RPE by gentle aspiration following a prior local vitrectomy and bleb detachment. Comparison was made between small and large areas of RPE removal, or only slight pressure to the retina without removal of RPE. RESULTS: Removal of the RPE layer causes transient leakage of vascular fluid into the subretinal space for at least a week after surgery and loss of perfusion in the underlying choroidal vessels and choriocapillaris at the debridement site. This reduction in local choroidal blood flow can occur within 15 min after RPE debridement and can be transient or permanent. Histology indicates that permanent changes are due to fibroblastic infiltration that compresses the choroidal vessels. Permanent changes tend to occur after removal of relatively large areas of RPE. The removal of small areas of RPE or slight pressure on the retina causes a transient loss of local choroidal perfusion, and fibroblastic infiltration into the choroid does not occur. CONCLUSION: Removal of the RPE causes changes throughout the underlying choroid. It reduces the circulation in the large choroidal vessels as well as the choriocapillaris. If large areas of RPE are removed, this choroidal non-perfusion can be permanent due to fibroblastic infiltration. Small areas of RPE removal or slight pressure on the retina lead to only transient reduction of the choroidal flow. The rapidity with which the choroidal blood flow can be reduced implies a reflex mechanism that responds to sudden RPE pressure and/or trauma. These changes are best observed with ICG angiography. PMID- 12883909 TI - Corneal shrinkage induced by nonmechanical Q-switched erbium:YAG laser trephination for penetrating keratoplasty in porcine eyes. AB - PURPOSE: To assess the degree of corneal diameter shrinkage induced by Q-switched mid-infrared laser corneal trephination for penetrating keratoplasty in an experimental model. METHODS: Corneal trephination was performed in 80 enucleated porcine eyes fixed in a holder centered on an automated globe rotation device, by Q-switched (2.94 microm) Er:YAG laser along open masks. Four types of masks were used to protect the underlying corneal tissue: metal masks (donor and recipient) and ceramic masks (donor and recipient). Two spot diameters (0.65 mm and 0.96 mm) were combined with two energy settings (40 mJ/pulse and 50 mJ/pulse) for each of the masks used. Repetition rate was fixed at 5 Hz. Diameters of donor buttons/recipient beds (horizontal and vertical) were measured immediately after the trephination and compared to the given mask size. RESULTS: Minimum corneal shrinkage was found in the recipient metal mask group (mean +/- SD=0.3+/-0.4%) with 50 mJ pulse energy and 0.65 mm spot diameter (in the horizontal diameter), while the maximum shrinkage (5.3+/-2.8%) was found in the donor metal mask group with 50 mJ pulse energy and 0.96 mm spot diameter. Corneal shrinkage was less pronounced in recipient beds than in donor buttons (P<0.01). The differences in shrinkage between the use of ceramic and metal masks were insignificant (P>0.05). Mean induced corneal diameter discrepancies between the donor button and the recipient bed (with metal and ceramic masks) were 2.5% and 2.5% in vertical diameter and 3.4% and 2.4% in horizontal diameter. CONCLUSIONS: The Q-switched Er:YAG laser experimental corneal trephination for penetrating keratoplasty may induce minor degrees of corneal diameter shrinkage in donor buttons and recipient openings. Oversizing of donor masks by 0.25-0.35 mm (i.e. 3-4% of graft size) may be a valid option to avoid refractive consequences. PMID- 12883910 TI - Topographic change in the central macula coupled with contrast sensitivity loss in diabetic pregnancy. AB - PURPOSE: To study macular topography and contrast sensitivity (CS) in diabetic and nondiabetic women during pregnancy and post partum. METHODS: A prospective study of 46 pregnant women with insulin-dependent diabetes and 11 nondiabetic pregnant controls. Macular surface topography was analyzed by Heidelberg Retinal Tomograph. Volume above the reference plane (VARP) was measured with 1.0-, 1.5-, 2.0-, and 3.0-mm-diameter circles. CS was measured with the Vistech 6500 Contrast Test System. RESULTS: The diabetic women had greater VARP than the controls measured with the 1.5-mm diameter circle. In diabetic women, the mean VARP was 0.084+/-0.064 mm(3) (mean +/- SD) in the first trimester, 0.080 +/- 0.056 mm(3) in the third trimester, and 0.087 +/- 0.067 mm(3) 3 months post partum compared with the values of 0.069+/-0.043, 0.054+/-0.024, and 0.036+/-0.020 mm(3) in the controls ( P=0.036 between groups). In diabetic women requiring laser treatment, the difference from controls was more significant ( P<0.001). CS at 3 cpd and 6.0 cpd was lower in diabetic women than in controls throughout pregnancy and post partum ( P=0.012 and P=0.043). A statistically significant negative correlation appeared between macular topography and CS during the third trimester; between cpd 6 and VARP 1.5 mm ( r=-0.471, P=0.001), and between cpd 6 and VARP 2.0 mm ( r=-0.446, P=0.002). CONCLUSIONS: In the diabetic women, especially in those with clear progression of retinopathy during pregnancy and post-partum, the macula seemed to be slightly more elevated than in the controls, and CS was lower at mid range spatial frequencies. CS loss in the diabetic women correlated with macular elevation during the third trimester. PMID- 12883911 TI - Visual acuity testing in diabetic subjects: the decimal progression chart versus the Freiburg visual acuity test. AB - PURPOSE: To study reproducibility and biological variation of visual acuity in diabetic maculopathy, using two different visual acuity tests, the decimal progression chart and the Freiburg visual acuity test. METHODS: Twenty-two eyes in 11 diabetic subjects were examined several times within a 12-month period using both visual acuity tests. The most commonly used visual acuity test in Denmark (the decimal progression chart) was compared to the Freiburg visual acuity test (automated testing) in a paired study. RESULTS: Correlation analysis revealed agreement between the two methods (r(2)=0.79; slope=0.82; y-axis intercept=0.01). The mean visual acuity was found to be 15% higher (P<0.0001) with the decimal progression chart than with the Freiburg visual acuity test. The reproducibility was the same in both tests (coefficient of variation: 12% for each test); however, the variation within the 12-month examination period differed significantly. The coefficient of variation was 17% using the decimal progression chart, 35% with the Freiburg visual acuity test. CONCLUSION: The reproducibility of the two visual acuity tests is comparable under optimal testing conditions in diabetic subjects with macular oedema. However, it appears that the Freiburg visual acuity test is significantly better for detection of biological variation. PMID- 12883912 TI - Choroidal neovascularization after laser-assisted in situ keratomileusis following penetrating keratoplasty. AB - PURPOSE: To describe a case of choroidal neovascularization (CNV) after laser in situ keratomileusis (LASIK) following penetrating keratoplasty (PK). METHODS: Case report. RESULTS: A 30-year-old man underwent PK in both eyes for bilateral keratoconus in 1997. Two years later, best-corrected visual acuity (BCVA) was 20/40 (-6=-4.50 x 170 degrees ) in RE and 20/20 (-1.50=-0.50 x 90 degrees ) in LE. To reduce the anisometropic defect, LASIK was performed in RE. After surgery, the refractive defect in RE reduced to -1.75 x 125 degrees and BCVA improved to 20/25. Six months after LASIK the patient presented loss of vision and metamorphopsia in RE due to choroidal neovascularization. BCVA was reduced to 20/200. Photodynamic therapy was performed in RE; 1 year later BCVA was stable at 20/200. CONCLUSION: Vitreoretinal complications after LASIK occur rarely. The potential relationship between CNV and LASIK is discussed. PMID- 12883913 TI - Hydrophobic acrylic versus heparin surface-modified polymethylmethacrylate intraocular lens: a biocompatibility study. AB - BACKGROUND: The implant of intraocular lenses (IOLs) following cataract surgery induces a foreign-body reaction to the IOL and a response on the part of the lens epithelial cells (LECs). The purpose of this study was to compare these aspects after the implantation of two different IOL materials. METHODS: Thirty-six cataract patients were randomised to receive two different foldable lens: an acrylic hydrophobic IOL (Acrysof MA30BA) and a heparin surface-modified (HSM) polymethylmethacrylate IOL (Pharmacia & Upjohn 809C) after phacoemulsification. Slit-lamp biomicroscopy with specular technique was used to assess the inflammatory cell adhesion on the anterior IOL surface, anterior capsule opacification (ACO) and membrane growth from the rhexis edge at 7, 30, 90, 180 and 360 days after surgery. RESULTS: The 809C group showed a higher percentage of patients with slight inflammatory cell adhesion on the anterior cell surface and a higher small cellular density during the whole follow-up period. The epithelioid cell response was greater in the 809C group than the Acrysof group but the difference between the two groups was not statistically significant. The ACO increased during the follow-up in both groups but was significantly higher in the 809C group. CONCLUSION: Acrysof lenses are more biocompatible than HSM IOLs, showing a lower grade of inflammatory cell adhesion and ACO. The implantation of these lenses may be particularly indicated in patients with pathologies predisposing to blood-aqueous barrier damage. PMID- 12883914 TI - Results and complications of hydrophobic acrylic vs PMMA posterior chamber lenses in children under 17 years of age. AB - PURPOSE: To compare results and complications of implantation of hydrophobic acrylic foldable intraocular lenses in children with those of polymethylmethacrylate (PMMA) intraocular lenses. METHODS: In a retrospective study, we analyzed results of cataract surgery with posterior chamber lens implantation in 30 eyes of 30 patients aged 1-16 years. In 10 eyes, acrylic (Alcon AcrySof) intraocular lenses, and in 20 eyes, single-piece PMMA posterior chamber lenses were implanted. Indications for cataract surgery (blunt or penetrating trauma, zonular cataract, cataracta polaris posterior, posterior lenticonus) and mean age at implantation were comparable in the two groups. Mean patient age at surgery was 8.6+/-4.6 years (range 3-16 years) for the acrylic vs 6.3+/-4.3 years (range 1-16 years) for the PMMA group. Mean follow-up was 1.0+/ 0.7 years (range 0.1-2.2 years) in the acrylic group and 1.8+/-1.5 years (range 0.1-5.7 years) in the PMMA group. Primary anterior vitrectomy was performed in 7 eyes in the PMMA group and in 3 eyes in the acrylic group. In addition, one additional posterior capsulorhexis without anterior vitrectomy was performed in each group. Primary outcome measure was the occurrence of postoperative "complications" (fibrin, synechiae, posterior capsular opacification). For statistical evaluation, the Fisher exact test was used. RESULTS: When evaluating all complications together (at least one complication vs no complication), there were significantly less complications in the acrylic group (2 of 10 vs 15 of 20; p=0.007. For early complications (postoperative fibrin, synechiae) the difference was also significant (1 of 10 in the acrylic vs 11 of 20 in the PMMA group; p=0.02). The rate of posterior capsular opacification necessitating YAG capsulotomy was lower in the acrylic group (1 of 10 eyes) than in the PMMA group (7 of 20 eyes), but the difference did not reach statistical significance ( p=0.67). The postoperative time point of YAG capsulotomy was 21 months in the acrylic group and 19+/-10 months (range 6-33 months) in the PMMA group. IOL dislocation was not observed in any of the patients. Postoperative visual acuity was comparable in the two groups: 0.57+/-0.35 (0.03-1.0) in the acrylic vs 0.39+/ 0.34 (0.001-0.9) in the PMMA group ( p=0.83). CONCLUSIONS: Implantation of hydrophobic acrylic intraocular lenses in the capsular bag in children may be associated with less postoperative complications compared with implantation of PMMA lenses. This appears also to be true in children under age 6 years. The visual results seem comparable and correspond mainly to the underlying ocular pathology. PMID- 12883915 TI - Prevalence and risk factors for epiretinal membranes in a Japanese population: the Hisayama study. AB - PURPOSE: To examine the prevalence and risk factors for epiretinal membranes (ERMs) in a sample Japanese population. METHODS: In 1998 a cross-sectional community survey was conducted among residents of Hisayama. A total of 688 men and 1087 women living in Hisayama, Japan, aged 40 years or older consented to participate in the study. Each participant underwent a comprehensive physical examination that included an ophthalmic examination. The presence of ERMs was judged from grading based on fundus examination using indirect ophthalmoscopy, slit lamp examination, and color fundus photographs. This study used non stereoscopic 45 degrees fundus photographs to grade ERMs, whereas the other population-based studies used 30 degrees stereoscopic fundus photographs, which might explain some differences in prevalence of ERMs. Multiple logistic regression analysis was performed on the cross-sectional data to determine the risk factors for ERMs. The following ten possible risk factors were used: age; gender; hypertension; diabetes; serum cholesterol; serum HDL cholesterol; serum triglycerides; smoking habits; alcohol intake; and body mass index. RESULTS: The prevalence of ERMs was 4.0%, and increased with age. The ERMs were more prevalent in women (4.3%) than in men (3.5%). Multiple logistic regression analysis showed that age and serum cholesterol significantly associated with ERMs. CONCLUSIONS: This study suggests that ERMs are less common in the Japanese population than in similar populations in Western countries, and that hypercholesterolemia is a relevant risk factor for ERMs. PMID- 12883916 TI - Macular pigment density in healthy subjects quantified with a modified confocal scanning laser ophthalmoscope. AB - BACKGROUND: In the industrialized world age-related macular degeneration (ARMD) is the leading cause for legal blindness beyond the age of 65 years. Recent studies indicate that the density of the macular pigment (MP) may play a central role in the development and progression of age-related maculopathy (ARM). We present results of measurements of macular pigment density (MPD) with a modified confocal scanning laser ophthalmoscope and compare macular pigment density values obtained from reflectance and autofluorescence images in healthy subjects. METHODS: A modified confocal scanning laser ophthalmoscope (Heidelberg Engineering, Heidelberg) was optimised for fundus reflectance and autofluorescence images at 488 nm and 514 nm. For autofluorescence imaging a band pass barrier filter with a short wavelength cut off at 530 nm was used. MPD maps were obtained, and MPD was evaluated within two degrees around the foveal center. In this study we included 109 healthy subjects aged from 18-75 years. RESULTS: Average MPD ranged between 0.16 density units (D.U.) and 0.22 D.U. depending on the techniques used for the measurement. Using the reflectance method the mean MPD was 0.16+/-0.06 D.U., whereas a MPD of 0.22+/-0.07 D.U. resulted from autofluorescence images with the 530-nm barrier filter. CONCLUSIONS: This study demonstrates the good feasibility of MPD measurements with a modified confocal SLO. In agreement with previous studies we found higher macular pigment density values calculated from autofluorescence images than from reflectance images. We could not find any differences in MPD related to age and gender. Prospective longitudinal studies to verify the influence of MPD on the development and progression of ARM are needed. PMID- 12883917 TI - Pathophysiology of tics and Tourette syndrome. AB - Tics are involuntary movements that can affect one or more muscles producing simple or complex movements. Blink reflex and startle reflex studies disclose an increased excitability of brainstem interneurons. Analysis of voluntary movement shows that when advance visual information is reduced, patients with tics and Tourette syndrome become progressively slower in completing motor sequences. Sensorimotor integration is abnormally processed. Studies of the contingent negative variation demonstrate abnormalities of movement preparation and the investigation of premotor potentials shows that in some patients tics are not preceded by a normal premotor potential. Magnetic stimulation studies demonstrate an increased excitability of cortical motor cortex. Functional MRI, PET and SPECT studies show abnormal activation of cortical and subcortical areas. Dysfunction of basal ganglia-thalamo-cortical projections affects sensorimotor, language and limbic cortical circuits, and may explain why patients with Tourette syndrome have difficulty in inhibiting unwanted behaviors and impulses. PMID- 12883918 TI - Anti-inflammatory therapy in Alzheimer's disease: is hope still alive? AB - Based on observations from neuropathology, epidemiology, and in vitro and animal experiments, the inflammatory component of Alzheimer's disease (AD) has been considered a compelling target for therapeutic intervention. However, a summary of all published trial reports to date suggests that AD patients do not benefit from treatment with anti-inflammatory drugs. In this brief review, we try to reconcile these sobering trial results with recent observations from basic research and epidemiology that continue to strengthen the idea that inflammatory mechanisms play an important role in the pathogenesis of AD. We review the possibilities that (1) not all components of the inflammatory response in AD are detrimental, (2) beneficial effects of anti-inflammatory drugs may not be mediated by inflammatory pathways, and (3) the timing of the intervention should be in the earliest stages of the pathogenesis of AD, perhaps even before the first symptoms emerge. We conclude that studies on primary prevention of AD are the logical next step in testing the inflammatory hypothesis of AD. PMID- 12883919 TI - An unusual family with multiple movement disorders. AB - Multiple movement disorders presenting in the same family are rare. We present an unusual family where generalized dystonia, Huntington's disease, progressive supranuclear palsy and secondary paroxysmal dyskinesia co-exist. The index case presented with young-onset dystonia and tested negative for the DYT1 gene deletion. Her father was similarly affected. The father's brother (paternal uncle of the index) also had abnormal movements-a mixture of chorea and dystonia-and tested positive for the HD expansion. His son had secondary paroxysmal dyskinesia, and tested negative for the HD expansion. The index case and her father were also negative for the HD expansion. A paternal aunt of two of the cases had a clinical diagnosis of progressive supranuclear palsy. Dystonia is known to be a genetically heterogeneous condition. The co-existence of inherited generalized dystonia with other movement disorders may provide clues to its genetic localization. PMID- 12883920 TI - Cerebral microdialysis as a monitoring method in subarachnoid hemorrhage patients, and correlation with clinical events--a systematic review. AB - BACKGROUND: One of the goals in treating subarachnoid hemorrhage patients is to prevent or minimize vasospasm-induced ischemia. Intracerebral microdialysis is a rapidly developing tool to monitor physiological and pathophysiological changes in chemical processes associated with ischemia. OBJECTIVE: To determine the diagnostic accuracy of microdialysis in detecting ischemia or ischemic events in patients with subarachnoid hemorrhage. METHODS: A systematic review of clinical studies regarding microdialysis as a monitoring method in patients in the acute stage of subarachnoid hemorrhage was conducted by performing a MEDLINE search using the terms "subarachnoid hemorrhage", "brain ischemia", "intracranial aneurysm", "cerebrovascular accident" and "stroke". These were combined with a search on "microdialysis". The methodological quality of the studies was assessed independently by two reviewers. For each study the grades of recommendation were determined. RESULTS: The search yielded 73 publications of which 13 studies were analysed. The methodological quality of the studies was low,with only 3 studies fulfilling more than 50% of the criteria. Consequently the level of evidence was low. Of the internal validity criteria the characteristics of study populations were similar in most of the studies. The technical assessment varied considerably. A quantitative analysis (meta-analysis) could not be performed because of lack of sufficient data. A qualitative analysis yielded a positive impression with regard to the diagnostic accuracy of microdialysis in detecting ischemic events. CONCLUSION: There is a positive tendency to use microdialysis as a diagnostic tool for monitoring of ischemic events in subarachnoid hemorrhage patients. However, there is insufficient evidence for the routine use of microdialysis. PMID- 12883921 TI - Criteria for diagnosis of pure neural leprosy. AB - The clinical diagnosis of pure neural leprosy (PNL) remains a public health care problem mainly because skin lesions-the cardinal features of leprosy-are always absent.Moreover, the identification of the leprosy bacillus is not easily achieved even when a nerve biopsy can be performed. In an attempt to reach a reliable PNL diagnosis in patients referred to our Leprosy Outpatient Clinic, this study employed a variety of criteria. The nerve biopsies performed on the 67 individuals whose clinical, neurological, and electrophysiological examination findings strongly suggested peripheral neuropathy were submitted to M. leprae identification via a polymerase chain reaction (PCR). Mononeuropathy multiplex was the most frequent clinical and electrophysiological pattern of nerve dysfunction, while sensory impairment occurred in 89% of all cases and motor dysfunction in 81%. Axonal neuropathy was the predominant electrophysiological finding, while the histopathological nerve study showed epithelioid granuloma in 14% of the patients, acid fast bacilli in 16%, and nonspecific inflammatory infiltrate and/or fibrosis in 39%. PCR for M. leprae was positive in 47% of the nerve biopsy samples (n=23). PCR, in conjunction with clinical and neurological examination results, can be a powerful tool in attempting to identify and confirm a PNL diagnosis. PMID- 12883922 TI - Early cerebral hemodynamic changes during passive movements and motor recovery after stroke. AB - Recovery from hemiplegia is a complex phenomenon that depends on various adaptive processes involving both the affected and the unaffected hemisphere. Our aim in this study was to investigate changes in cerebral perfusion in hemiplegic stroke patients during passive movements and their correlation with the subsequent motor recovery. The study included 30 patients with single, subcortical ischemic cerebral lesions. Within 14 days (range 8 to 14 days) from stroke onset, all patients were examined for the effects of passive elbow movements on cerebral blood flow in the middle cerebral arteries (MCAs) by means of bilateral transcranial Doppler (TCD) ultrasonography. On the same day as TCD assessment, they were also evaluated clinically with the Canadian Neurological Scale (CNS) and with Medical Research Council (MRC) scale for motor deficit of the affected arm. A clinical evaluation using the same scales was repeated after two months of motor rehabilitation therapy. We investigated the relationship between changes of Mean Flow Velocity (MFV) during passive movements and degree of recovery after stroke. The logistic regression procedure indicated that out of all factors considered as possibly related to a good clinical motor deficit recovery of the affected arm, evaluated by means of MRC, only the MFV percentage increase played a predictive role. In particular, for each additional point of contralateral MFV percentage increase during passive movement of the affected arm, the relative probability of good clinical recovery increased 5.68 times (95% CI=1.76-18.40; p=0.004). Similar results were found when the clinical recovery was measured by means of the CNS (slope=0.40, p<0.001). Passive movements in hemiplegic stroke patients before clinical recovery elicit activation patterns that may be critical for the restoration of motor function.I n particular, early and consistent activation of the affected hemisphere, as detected with TCD, seems to predict the positive evolution of a motor deficit. PMID- 12883923 TI - Side effects of intravenous immunoglobulins in neurological autoimmune disorders- a prospective study. AB - The increased use of intravenous immunoglobulins (IVIg) in the treatment of neurological autoimmune diseases has led to more awareness of adverse reactions. We studied prospectively the side effects of IVIg during 84 treatment courses with a total of 341 infusions under routine clinical conditions. Mild reactions were common. Headache was noted most often, occurring during 30% of treatment courses. There were three severe adverse events (3.6% of all treatment courses) that led to discontinuation of the treatment, namely thrombosis of the jugular vein, allergic reaction and retrosternal pressure. Significant changes in laboratory findings were seen for leucocytes, erythrocytes, haematocrit, haemoglobin, ALAT and ASAT. None of these changes were clinically relevant. The elevation of liver enzymes was dependent on the IVIg preparation used, while there was no association with the underlying disease, age, or gender of the patient. In conclusion, this prospective study confirms the high frequency of mild, self-limited side effects of IVIg. Elevation of liver enzymes may possibly be associated with certain IVIg preparations. Bearing these complications in mind, this prospective study supports the notion that IVIg can generally be regarded as safe, leading to severe adverse events during only 3 (0.9%) of 341 infusions (or 3 of 84 treatment courses, 3.6 %). However, careful monitoring for severe side effects remains mandatory, and we propose that laboratory findings like full blood count, renal and liver function should be monitored routinely. PMID- 12883924 TI - Dual dopamine agonist treatment in Parkinson's disease. AB - We studied the effectiveness of cabergoline as an adjunct in patients with early Parkinson's disease receiving another dopamine agonist (pramipexole or ropinirole), at the maximal permitted dose. The study enrolled 47 patients: 35 had motor fluctuations and were receiving a dopamine agonist with levodopa; and 12, without motor fluctuations, were receiving a dopamine agonist without levodopa. These medications had in all cases failed to achieve adequate symptom control. In the 35 patients with motor fluctuations, when cabergoline was added to therapy, the time spent in the OFF period decreased by 65.6% (from 3.14+/-1.11 to 1.08+/-1.07 hours p<0.0001); and the Unified Parkinson's Disease Rating Scale (UPDRS) motor score decreased by 19.24% (from 41.68+/-12.6 to 33.66+/-10.22; (p<0.0001) during the OFF condition, and by 7.11% (from 17.01+/-6.63 to 15.8+/ 7.22; p<0.001) during the ON condition. Nocturnal akinesia improved in all the patients except one. In the 12 patients without motor fluctuations, when cabergoline was added, the UPDRS score improved by 34.4% (from 23.5+/-5.3 to 15.5+/-4.7). This open study shows that dual dopamine agonist therapy can be useful in the symptomatic treatment of patients with early or more advanced Parkinson's disease receiving therapy with or without levodopa. PMID- 12883925 TI - Metabolic correlates of behavioral and affective disturbances in frontal lobe pathologies. AB - OBJECTIVE: Although previous studies have shown that the human frontal cortex is involved in the experience of emotions as well as in social behavior, data regarding the exact anatomical substrates of behavioral and affective deficits in frontal lobe pathologies are still scarce. The aim of this study was to investigate the metabolic correlates of these deficits in a group of non-selected consecutive patients with frontal lobe lesions. PATIENTS AND METHODS: Clinicometabolic correlations between several emotional and social parameters and metabolic patterns in the frontal cortex and amygdala were investigated in 32 patients with frontal lobe pathologies. The behavioral disturbances were evaluated using the Lhermitte's informant questionnaire. Regional cerebral glucose metabolism was measured with [(18)F] fluorodeoxyglucose and high resolution positron emission tomography. Statistical analysis was performed using both single variable correlation and multiple regression analyses. RESULTS: Both single variable and multivariate analyses demonstrate that decreased regional glucose metabolism in the right medial area 10 was associated with apathy. There were also significant negative relationships between metabolism in the right orbitofrontal cortex and stereotypy and indifference to rules. Impulsiveness, personality disturbances and loss of emotional control were associated with decreased metabolism in the left amygdala. CONCLUSIONS: In terms of clinicometabolic correlations, the present data support the implication of different functional anatomic systems in frontal lobe-related behavioral and affective disturbances. In particular, they imply that the classically described symptoms of impaired behavioral control may be related to right orbitofrontal cortex hypometabolism whereas impaired regulation of emotions may result from a functional damage of the left amygdala. PMID- 12883926 TI - Cyclophosphamide is effective in stabilizing rapidly deteriorating secondary progressive multiple sclerosis. AB - The safety and efficacy of pulse cyclophosphamide (CTX) therapy was investigated in patients with very active secondary progressive multiple sclerosis, characterized by frequent relapses and rapid disability progression. For this purpose the clinical and MRI effects were assessed. Sixteen patients, 11 female and 5 male, were experiencing rapidly deteriorating disease, characterized by frequent and severe relapses as well as rapid progression (defined by an increase of more than 1 EDSS point in a period of 1 year). Mean relapse rate in the two years preceding CTX therapy was 3.0 +/-1.4. Mean EDSS was 4.0+/-1.4 one year before therapy and 5.6+/-1.0 at study entry. Treatment consisted in administration of high dose intravenous CTX every four weeks for one year and then every eight weeks for an additional twelve months. CTX dose was tailored to the patient's white blood cell response, and ranged from 800 to 1,200 mg/m(2) body surface. MRI was performed before therapy and then at 12 (Y1) and 24 (Y2) months. Eight patients with similar clinical features constituted a control group. CTX therapy was safe and well tolerated, and no severe side effects were observed. The EDSS decreased to 4.3+/-1.6 at Y1 (Y0 vs.Y1: p< 0.001) and to 4.1+/ 1.6 at Y2 (Y0 vs.Y2: p< 0.001). Only four patients experienced relapses during the first year of therapy, while no relapses were observed during the second year of therapy. The mean relapse rate during therapy was 0.25 +/-0.45 (p< 0.0001). No increase in T2 lesion load was observed over the two years. A significant clinical and MRI deterioration was observed in the control group. Therapy with pulse CTX was able to stop disease activity and progression in patients with rapidly evolving secondary-progressive MS. PMID- 12883927 TI - Epileptic seizure after stroke in Chinese patients. AB - This was a hospital-based cohort study aiming at determining the occurrence rate of post-stroke seizures and the associated risk factors. From 27 July 1996 to 16 June 1998, the first 1000 consecutive patients in the acute stroke registry were retrospectively reviewed for one year after acute stroke to identify seizure occurrence. The demographic data, seizure onset time, seizure type, drug treatment, response to medication, electroencephalogram findings and cranial computed tomogram findings were collected. Thirty-four patients (3.4%) developed seizure within one year after acute stroke. Univariate analysis revealed that male, age greater than 65 years, total anterior circulation infarction, partial anterior circulation infarction, cortical location and large lesion were significantly associated with post-stroke seizure while multivariate analysis showed that only male (adjusted OR 3.21, p<0.01) and cortical location (adjusted OR 3.83, p<0.05) were significant independent risk factors. Fifty-six percent of early seizures were partial type whereas 72% of late seizures were generalized tonic-clonic type of undetermined onset. Seizures occurred in 3.4% of patients within one year after the onset of stroke. This percentage of seizure occurrence and associated risk factors were similar to other studies. However, intracerebral and subarachnoid haemorrhage were not shown to be risk factors in our study. PMID- 12883928 TI - Giant cell arteritis of the occipital arteries--a prospective color coded duplex sonography study in 78 patients. AB - CONTEXT: Occipital headache and nuchal pain may indicate the involvement of the occipital arteries (OCCA) in temporal arteritis (TA). Recently high resolution color coded sonography (CCDS) has greatly improved the imaging of small lumen arteries. OBJECTIVE: The aim of the present study was the demonstration of TA, of the OCCA in comparison with the superficial temporal artery (STA) by means of CCDS in patients with nuchal and occipital pain suspected of suffering from TA. DESIGN: Prospective study of 78 patients comparing CCDS findings of the OCCA and of the STA with the clinical diagnosis and the biopsy results. RESULTS: 27 patients received the diagnosis TA; there were 51 other diagnoses. CCDS of the OCCA reached a sensitivity in diagnosis of 63 % and a specificity of 100 % and in histology of 65 % and 100% respectively. CCDS of the STA reached a diagnostic sensitivity of 78 % and a specificity of 94% and of a histological sensitivity of 77 % and specificity of 82 %. Reversibility of CCDS abnormalities was monitored in 5 patients over a period of 13 to 42 days. CONCLUSIONS: Involvement of the OCCA in TA patients is a frequent finding and may be the only pathological phenomenon in some patients with nuchal pain, occipital headache and occipital scalp tenderness. CCDS of the STA and OCCA contributed to the diagnosis of TA with a high rate of perivascular hypoechogenic abnormalities (stenoses and occlusions) and a low rate of these abnormalities in the control patients. However, CCDS cannot differentiate between inflammatory and degenerative artery disease and has limitations concerning spatial resolution. Before CCDS may replace biopsy in clinical practice the accuracy of the criteria recommended above should be tested in larger groups of patients. PMID- 12883929 TI - Deficits of predictive grip force control during object manipulation in acute stroke. AB - Anticipatory grip force adjustments when lifting, holding and performing vertical point-to-point movements with a hand-held object were analysed in 11 patients with deficits of fine manual motor performance due to acute ischemic stroke. All patients had mild to moderate paresis and sensory deficits of the affected hand. Grip forces used to stabilise the object in the hand, accelerations of the object and movement-induced loads were measured. Compared with controls, patients produced markedly increased grip forces when lifting, holding and moving the hand held object. The ratio between grip force and the actual load,which is considered to be a sensitive measure of force efficiency, was significantly elevated in stroke patients indicating a strategic generalisation of grip force increase when cerebral sensorimotor areas are functionally impaired. The temporal coupling between grip and load force profiles revealed only selective impairments during the lifting and movement tasks of stroke patients. The time to reach maximum grip force was prolonged and there were greater time lags between grip and load force maxima during the lifting movements. When healthy controls performed vertical movements with the hand-held object grip force increased early in upward and late in downward movements and grip and load force maxima coincided closely in time. The time lags between maximum grip and load forces were similar for vertical movements performed by patients and controls. However, the time lags between grip force and acceleration onset were larger for upward and smaller for downward movements performed by stroke patients. These findings indicate impaired prediction of the inertial load profiles arising from voluntary arm movements with a hand-held object in acute stroke. PMID- 12883930 TI - Effects of a short outpatient rehabilitation treatment on disability of multiple sclerosis patients--a randomised controlled trial. AB - It is well known that neurorehabilitation can reduce disability or improve handicap of people with multiple sclerosis (MS). The aim of this study was to evaluate the effectiveness of a short period (6 weeks) of a tailored, individualised outpatient rehabilitation program in people with progressive MS. A randomised-controlled trial was undertaken in patients with primary and secondary progressive MS referred to the Centro Sclerosi Multipla of Catania. One hundred and eleven patients were assessed at baseline and at 12 weeks with validated measures of disability (Functional Independence Measure (FIM)) and impairment (Expanded Disability Status Scale (EDSS) and Functional Systems Scale). Of the 111, 58 were randomly assigned to the treatment group and 53 to the control group. All patients had been previously trained in a home exercise program. Both groups were well matched for age, sex, disease duration and severity, disability and quality of life (Short Form-36). At the end of 6 weeks patients allocated to the rehabilitation treatment group showed significant improvement in their level of disability compared with the control group,while the level of impairment did not change. Thirty-two patients of the treatment group and four of the control group improved on the FIM by two or more steps at 12 weeks (p<0.0001). An improvement by 1 EDSS step occurred in only two patients of the treatment group and in one patient of the control group. Benefits were maintained for a further six weeks. This study demonstrates that a short outpatient rehabilitation treatment improves disability of MS patients, without changing their impairment and confirms the effectiveness of rehabilitation in people with MS. PMID- 12883931 TI - Diffuse white matter lesions in a case of herpes simplex encephalitis. PMID- 12883932 TI - Streptococcus suis meningitis and septicemia contracted from a wild boar in Germany. PMID- 12883933 TI - Clinically definite multiple sclerosis after radiological Schilder-like onset. PMID- 12883934 TI - Myelin basic protein in multiple sclerosis and other neurological disorders. PMID- 12883935 TI - Spastic paraparesis without optic atrophy after occupational methanol exposure. PMID- 12883936 TI - An Italian family with Nasu-Hakola disease. PMID- 12883937 TI - Acute oropharyngeal palsy associated with anti-GM1b IgG antibody. PMID- 12883938 TI - Ice pack test for myasthenia gravis. A simple, noninvasive and safe diagnostic method. PMID- 12883939 TI - Essential palatal myoclonus in monozygotic male twins. PMID- 12883940 TI - Intestinal pseudobstruction as presenting event of fatal cerivastatin-induced myopathy. PMID- 12883941 TI - Derek Ernest Denny-Brown (1901-1981). PMID- 12883944 TI - Significance of micro-geographical population structure in forensic cases: a bayesian exploration. AB - We studied the influence of population structure at the microgeographical level on the analysis of forensic cases. A total of nine autosomal STRs and seven Y STRs were analyzed in the general mixed population and in two relatively isolated valleys of Cantabria, a region in Northern Spain. Statistically significant differences existed in the frequency distribution of four autosomal STRs, with an overall Fst value of 0.3%. A simulation of virtual trio cases revealed that it did not have a practical influence on the analysis of paternity disputes. Significant differences also existed in most Y-STRs, with an overall Fst value of 3%. Thus, using the general database instead of the specific valley database resulted in 5-fold or higher overestimation of the likelihood ratio of matching in up to 30% of cases. A bayesian analysis revealed that this had a significant impact on the estimation of the probability of identity in scenarios of low "a priori" odds of suspicion. PMID- 12883945 TI - CEN plasmid segregation is destabilized by tethered determinants of Ty 5 integration specificity: a role for double-strand breaks in CEN antagonism. AB - The yeast retrotransposon Ty 5 integrates preferentially into heterochromatin at the telomeres and HM loci. Target specificity is mediated by a six amino acid sequence motif (the targeting domain, TD) of integrase that interacts with Sir4p, a structural component of heterochromatin. When tethered to CEN plasmids as part of a Gal4p DNA binding domain (GBD) fusion protein, TD destabilizes plasmid segregation in a manner similar to that observed for CEN + HM or CEN +TEL antagonism. This instability is caused by the ability of TD to nucleate components of heterochromatin on the CEN plasmid, because CEN +TD antagonism is abrogated by sir2, sir3 and sir4 mutations and by TD mutations that prevent interaction with Sir4p. In strains that acquire resistance to CEN +TD antagonism, the CEN plasmid has either recombined with a 2 mu plasmid or sustained deletions in sequences required to bind GBD-TD. CEN +TD and CEN + HM antagonism is exacerbated by mutations in components of the Ku-mediated non-homologous end joining pathway. These observations suggest that CEN antagonism is caused by DNA breaks that result from competition between CEN - and Sir-specific segregation pathways. PMID- 12883946 TI - Analysis of detached human kinetochores. AB - A method has recently been established for inducing the physical detachment of kinetochores from chromosomes in human HeLa cells, and was used in the studies reported here to investigate the organization and function of dissociated HeLa kinetochores. Immunofluorescence labeling demonstrated that the detached HeLa kinetochores were relatively intact, with the number of detached kinetochores being only moderately more than the diploid number of chromosomes in HeLa cells. In addition, the detached kinetochores could be labeled with antibodies specific for the inner kinetochore plate, outer kinetochore, and subjacent centromeric heterochromatin. A functional assay demonstrated that detached kinetochores retained the capacity to activate the spindle checkpoint, leading to metaphase arrest. Analysis of kinetochore DNA indicated that it consisted primarily of DNA fragments of 130-160 kb in size, while the remainder of the chromosomes were sheared into much smaller fragments during the kinetochore detachment event. Further analysis of kinetochore DNA indicated that it was first cleaved into high molecular weight DNA (>200 kb) fragments during the initial stages of the kinetochore detachment process, and then underwent further maturation following nuclear envelope breakdown to give rise to the 130-160 kb fragment in detached kinetochores. Collectively, these data indicate that detached human kinetochores will be a useful system for investigating the organization, assembly, and function of human kinetochores. PMID- 12883947 TI - Localization of importin alpha (Rch1) at the plasma membrane and subcellular redistribution during lymphocyte activation. AB - Rch1 belongs to the importin alpha subfamily and works as an adapter between karyophilic proteins and the nuclear import machinery. Its level of expression varies among species and tissues, and depends on the state of cellular metabolism. In the present study we examined the level of expression of nuclear envelope and nuclear transport proteins (Rch1, importin beta, lamins A/C, lamin B, gp210, p62 and transportin) after human lymphocyte activation with phytohemagglutinin. We observed that the level of Rch1 increases dramatically, especially in larger lymphocytes, in response to activation. Moreover, using immunoelectron microscopy, this nuclear transport factor was found to be localized at the plasma membrane and also in tracks from the cytoplasm through the nuclear envelope into the nucleus. Similar localization was also observed in the human melanoma cell line A375. In addition, metabolic activation led to a redistribution of Rch1 from the cytoplasm to both the plasma membrane and the nuclear interior. These results suggest that, during lymphocyte activation, Rch1 may be involved in a signal transduction pathway that involves the shuttling of karyophilic proteins from the plasma membrane to the nucleus. PMID- 12883948 TI - Clonogenic survival of human keratinocytes and rodent fibroblasts after irradiation with 25 kV x-rays. AB - Low energy x-rays (E(ph) 90%. However production of active metalloproteinases by disc cells was relatively insensitive to pH, with activity at pH 6.3 not statistically different from that at pH 7.2. These findings indicate that exposure to acid conditions appears particularly deleterious for the disc matrix, as it inhibits the disc cells from synthesising functionally important molecules such as the sulphated GAGs but does not prevent the production of agents able to degrade matrix components. The low values of pH seen in some degenerate discs are thus likely to be involved in breakdown of the disc matrix. PMID- 12883963 TI - Associations among awareness of prognosis, hopefulness, and coping in patients with advanced cancer participating in phase I clinical trials. AB - GOALS: There is considerable debate concerning prognosis in patients with advanced cancer (advanced cancer patients, ACP), but no one has systematically examined the relationship between awareness of prognosis and hopefulness. In this study, the relationships among awareness of prognosis, hopefulness, and coping were examined in a selected group of ACP. PATIENTS AND METHODS: All subjects were enrolled in phase I trials. Information was obtained from subjects via structured, face-to-face interviews. Subjects provided demographic information, answered questions designed to ascertain their awareness of their prognosis, and completed the Functional Assessment of Cancer Therapy-General instrument (FACT G). RESULTS: A total of 179 consecutive subjects enrolled in phase I trials were interviewed. Awareness of prognosis was assessed using a semiquantitative scale. Willingness to provide responses to the question was significantly negatively correlated with hopefulness ( r=-0.306, P=0.030). Using items from the FACT-G, hopefulness and coping were positively correlated ( r=0.140, P=0.097). Female subjects were more realistic about their prognosis than men ( b=0.568, P=0.026). More educated patients had higher response rates to the prognosis questions (OR 1.62, P<0.001), and demonstrated a more realistic awareness of prognosis ( b=0.225, P=0.030). Overall, there was a strong association between coping status and responding to the prognosis questions ( P=0.033). Patients who reported being "not at all proud of coping" had a lower response rate than patients who reported better coping. DISCUSSION: Among the ACP who provided responses, many reported an unrealistic view of their prognosis. Having a more accurate view of prognosis in the face of terminal illness was associated with reduced hopefulness, which may be related to a poorer sense of coping. Finally, the prognosis question utilized in the study was problematic, led to a fairly poor response rate, and may be indicative of the many difficulties present in attempting to accurately assess ACPs' perceptions of their prognosis. Other methods, such as in-depth qualitative methods, may lead to more productive inquiry into ACPs' awareness of their prognosis. Further research into the accurate assessment of awareness of prognosis is needed. PMID- 12883964 TI - Time limitations and the challenge of providing infection-preventing dental care to hematopoietic stem-cell transplantation patients. AB - GOAL: Oral-dental infection foci should be eradicated before the ablative chemo radiotherapy regimen of hematopoietic stem cell transplantation (HSCT) commences. The rationale of oral-dental treatment is to prevent the future development and spread of infections in patients with compromised immune systems. This study aims to shed light on the challenges facing hospital dentists in the implementation of optimal oral-dental treatment prior to HSCT. PATIENTS AND METHODS: Data regarding the medical status and dental treatment needs before HSCT were retrieved from the files of 86 consecutive patients post-HSCT. The timing of the oral-dental examination was also recorded. MAIN RESULTS: Dental treatments required before the HSCT were mainly combinations of scaling, plastic fillings, and extractions (47.8%, 39.1%, and 19.5% of the patients respectively). Patients presented at the clinic for oral-dental examination an average of 20.65+/-16.82 days before HSCT with a median of 15 (quarter interval range 10-15) days. CONCLUSIONS: Our data indicate a dense distribution of dental needs preceding the ablative conditioning regimen for HSCT. These facts accentuate the vital need for cooperation between hospital dentists and treating physicians. PMID- 12883965 TI - Economic evaluation of antiaromatase agents in the second-line treatment of metastatic breast cancer. AB - GOALS OF WORK: An economic evaluation was conducted comparing anastrozole, exemestane, letrozole and megestrol for the second-line treatment of postmenopausal patients with hormone-sensitive metastatic breast cancer who had failed tamoxifen. METHODS: An economic model was developed based on data from phase III megestrol-controlled clinical trials of antiaromatase agents (AA) and estimates of resource utilization from both Statistic Canada's Population Health Model for breast cancer and expert opinion. MAIN RESULTS: In megestrol-controlled trials, anastrozole and exemestane equivalently improved survival compared to megestrol, while letrozole did not. Compared to megestrol, exemestane and anastrozole both cost Canadian $9000 per life-year gained, and letrozole saved Canadian $300 annually, with no life-year gain. Cost-effectiveness results were robust under sensitivity analysis testing. However, the model was dependent on any difference between AAs with respect to survival benefit and drug acquisition price. CONCLUSION: Based on available data and this cost-effectiveness analysis, exemestane and anastrozole are appropriate choices for second-line hormonal treatment of metastatic breast cancer. PMID- 12883966 TI - A bedside food cart as an alternate food service for acute and palliative oncological patients. AB - Patients with advanced cancer experience various problems with eating, and their meals should be tailored to meet their specific needs. Two methods of food service were compared in a shared acute oncology/palliative care unit; an electrical food cart allowing patients to select their food types and portions at the bedside, and a traditional food tray delivery service that relied on meals being prepared in a centralized kitchen and then delivered by tray. Over a 10-day period, lunch meals were delivered by food cart and supper meals via food trays. Twenty-seven out of 32 patients participated in the trial. Patients significantly preferred the food cart to the trays with respect to the timing and appeal of the meal, appropriateness of food types and food portions and the variety of the food choices. A food cart as used in this trial provides a more flexible and appropriate method of food delivery to in-patients in the oncology and palliative unit. Further studies should examine whether this translates to improved caloric intake and quality of life parameters. PMID- 12883967 TI - Renal involvement in human parvovirus B19 infection. PMID- 12883968 TI - Focal segmental membranoproliferative glomerulonephritis in children. AB - Eight patients with focal segmental membranoproliferative glomerulonephritis (FSMPGN) were followed for 5-16 years. Their urinary abnormalities were detected by school urinary screening in seven, and one patient presented with nephrotic syndrome. All but one patient were treated with alternate-day (ALD) prednisolone. With time, urinalysis became normal in six and two continued to have proteinuria with or without hematuria. Serum albumin, cholesterol, and creatinine levels were normal at the last follow-up. Serum C3 returned to normal levels in six, but remained persistently decreased in two. Mesangial proliferation and matrix changes in glomeruli without MPGN lesions were mild. Subendothelial and mesangial electron-dense deposits and deposits containing C3 along capillary walls and mesangium were observed. MPGN lesions and mesangial proliferation improved. No severe growth retardation was observed, but the duration and dosage of ALD prednisolone could be reduced further, since the patients with FSMPGN seemed to have an excellent prognosis. PMID- 12883969 TI - Natural history of voiding dysfunction. AB - Voiding dysfunction leads to daytime wetting, night-time wetting, and recurrent urinary tract infection (UTI). Our interdisciplinary center has been committed to treating children with dysfunctional voiding since 1992. We describe the long term follow-up of a large cohort of children using our approach to treatment. We reviewed the records of 199 children with symptoms of voiding dysfunction seen between March 1992 and December 1993. We contacted 98 parents and 51 patients by phone at least 6.5 years after initial presentation and asked about current symptoms, effective strategies, and satisfaction of care. Of the initial group of contacted parents, 81 (83%) were female with a median age of 7.8 years. Patients were followed in the clinic for a mean time of 1.6 years and a median of three visits. Of the 90 patients with daytime wetting, 82 (91%) reported complete resolution with a median time of resolution of 2.9 years. Parents felt that maturation was the most important factor leading to improvement. Of the patients that were directly contacted (41 females, 10 males, median age 15.2 years), all were dry and 86% denied any sense of urgency; 86% of parents and 87% of patients were highly satisfied with their care. Almost every child improved within 5 years of the initial evaluation. Only small fractions are still wet, are infected, or have urgency. Maturation seems to be the most important factor leading to improvement. Most parents and patients are satisfied with this form of treatment. PMID- 12883970 TI - Fetal renal pelvic dilatation--poor predictor of familial vesicoureteral reflux. AB - The sensitivity and positive predictive value (PPV) of fetal renal pelvic dilatation for detecting vesicoureteral reflux (VUR) can only be determined by performing voiding cystourethrography (VCUG) on all newborns in a study population. We sought to determine this for infants with a family history of reflux. From June 1989 to September 1996, 157 children (80 males) under 2 years with a family history of primary VUR in a first-degree relative had VCUG. VCUG results were compared with the fetal renal pelvic diameter on obstetric sonogram performed after 16 weeks gestation. Of the 157 infants, 33 (11 boys) had primary VUR. In those with a parent as the index case, the prevalence of reflux was 5% in boys and 35% in girls ( P=0.03). The largest renal pelvic diameter after 16 weeks was not discriminatory for VUR, but after 30 weeks gestation a 4 mm renal pelvis had a sensitivity of 33% and a PPV of 32%. The sensitivity was higher for grades 4 and 5 (75%) than for grades 1-3 reflux (17%), ( P=0.04). In conclusion, fetal renal pelvic diameter has a low sensitivity and poor predictive value for detecting VUR, but this improves a little after 30 weeks gestation. PMID- 12883971 TI - Kimura disease in a patient with renal allograft failure secondary to chronic rejection. AB - Recent studies suggest that CD4(+ )T helper 2 (Th2) cell proliferation and overexpression of Th2 cytokines may play an important role in the development of Kimura disease. Chronic rejection of a renal allograft by the indirect allo recognition pathway is also induced by Th2 cytokines. We report a 12-year-old boy who had presented with nephrotic syndrome 10 years previously. He was found to have focal segmental glomerulosclerosis, which was attributed to vesicoureteral reflux, and he underwent renal transplantation at the age of 5 years. Allograft dysfunction secondary to chronic rejection was noted by 2 years post transplant, after which continuous ambulatory peritoneal dialysis was instituted. After discontinuation of immunosuppressive therapy, he progressively developed peripheral eosinophilia and eczema, followed by cervical lymphadenopathy and then epitrochlear lymphadenopathy. Kimura disease was diagnosed on lymph node biopsy. Our patient demonstrated that Kimura disease can occur after renal allograft failure secondary to chronic rejection. Both disorders involve the Th2-dominant immune response, according to previous observations. PMID- 12883972 TI - Bone mineral density in children with myelomeningocele: effect of hydrochlorothiazide. AB - Children with myelomeningocele experience difficulty with ambulation, which leads to immobilization and secondary loss of bone mineral density (BMD). In addition, non-ambulatory myelomeningocele patients have higher urinary calcium losses than their ambulatory counterparts. Hydrochlorothiazide (HCTZ) is known to reduce urinary calcium loss and increase BMD in non-myelomeningocele patients with hypercalciuria. This study examines the effect of HCTZ on urinary calcium and BMD in non-ambulatory children with myelomeningocele. Thirteen of 20 non-ambulatory patients with myelomeningocele completed the year-long randomized double-blinded study (placebo = 7 and HCTZ = 6). Evaluation included electrolytes, PTH, osteocalcin, 1, 25-OH vitamin D, urinary pyridinolines/deoxypyridinolines (U(pyr/dpyr)), urinary calcium/creatinine (U(Ca/Cr)), and forearm BMD (dual X-ray absorptiometry). Follow-up electrolytes were obtained at 1-2, 6, and 12 months and U(Ca/Cr) and BMD was obtained again at 12 months. There were no initial differences between the placebo and HCTZ groups. U(Ca/Cr) decreased in the HCTZ group after treatment (0.20+/-0.09 vs. 0.04+/-0.02, p<0.05). However, forearm BMD ( z-scores) after 1 year remained unchanged in both the HCTZ (-5.95+/-0.98 to 5.86+/-0.92) and placebo (-7.19+/-0.69 to -6.67+/-0.63) groups. While use of HCTZ for 1 year did not affect BMD, it reduced urinary calcium excretion in non ambulatory children with myelomeningocele. PMID- 12883974 TI - Treatment of congenital nephrogenic diabetes insipidus by hydrochlorothiazide and cyclooxygenase-2 inhibitor. AB - A 1-month-old male infant presented with failure to thrive, polyuria, and severe hypernatremic dehydration. Based on family history, lack of response to vasopressin, and normal sonography of the urinary system, the diagnosis of congenital nephrogenic diabetes insipidus (cNDI) was established. The infant responded well to indomethacin in combination with hydrochlorothiazide (HCTZ), but quickly developed gastrointestinal bleeding. The substitution of indomethacin by amiloride and later by tolmetin was found to be ineffective. Treatment with HCTZ (3 mg/kg per day) and rofecoxib (1 mg/kg per day, both divided into three doses) combined with a low-salt formula resulted in a dramatic decrease in urinary free water losses. No side effects of the combination were noted. At age 8.5 months, the infant demonstrated catch-up growth and normal neurodevelopmental milestones. We conclude that the combination HCTZ/cyclooxygenase-2 inhibitor could be successfully used to treat infantile cNDI. PMID- 12883975 TI - Carotid ultrasonography for detection of vascular abnormalities in hypertensive children. AB - To determine whether hypertension and overweight status are associated with increased carotid intimal-medial thickness (cIMT) in children, vascular ultrasonography was performed in newly diagnosed hypertensive patients ( n=53) and normotensive controls ( n=33). Hypertensive subjects were identified either by referral or by systematic school-based hypertension screening. Hypertension was defined as blood pressure above the 95th percentile based on current Task Force criteria, and overweight was defined as body mass index (BMI) >25 kg/m(2). cIMT was assessed by high-resolution vascular ultrasonography of the distal common carotid artery. Hypertensive subjects had a higher cIMT than normotensive subjects (0.62 vs. 0.53 mm, P<0.00001). This difference remained significant after controlling for the effects of gender, race, age, height, weight, and BMI. Similarly, overweight subjects had a higher cIMT than normal-weight subjects (0.63 vs. 0.54 mm, P<0.0001). Subjects with both systolic and diastolic hypertension had higher cIMT than those with isolated systolic hypertension (0.67 vs. 0.60, P<0.05). cIMT showed significant positive pairwise correlation with age, height, weight, BMI, and systolic blood pressure. Among all clinical variables analyzed, cIMT was most strongly correlated with BMI ( r=0.53, P<0.001). These results provide further evidence that vasculopathy occurs in association with known cardiovascular risk factors such as hypertension and obesity during childhood. PMID- 12883976 TI - Low serum apolipoprotein AI levels in amyloidosis related to familial Mediterranean fever. AB - Amyloidosis (A) related to familial Mediterranean fever (FMF) causes serious morbidity and mortality in children. Our study evaluates serum levels of apolipoprotein (Apo) AI, AII, B, and E and Apo AII/AI ratios as a non-invasive diagnostic tool for amyloidosis in children with FMF and FMF-A. Results were compared with those of patients with childhood nephrotic syndrome (NS) and healthy children (controls). Significantly lower serum levels of Apo AI (90.20+/ 28.30 mg/dl) were documented in patients with FMF-A than in all other groups (FMF 126.89+/-51.07 mg/dl, NS 140.38+/-33.73 mg/dl, and controls 134.67+/-12.73 mg/dl) ( P<0.01). Diagnostic sensitivity, specificity, and predictive value for this test were 85%, 80%, and 85%, respectively. Apo AII/AI ratio results were essentially equal in all groups ( P>0.05). It is concluded that a decreased Apo AI serum level, but not Apo AII/AI ratio, is a useful, non-invasive test for the early diagnosis of FMF-A in children. PMID- 12883977 TI - Pharmacokinetics of ganciclovir in pediatric renal transplant recipients. AB - Ganciclovir (GCV) is effective in preventing and treating cytomegalovirus (CMV) infection in solid organ transplant recipients. The aims of the present study were to determine the pharmacokinetics of GCV administered intravenously (IV) and orally (p.o.) as pre-emptive anti-CMV therapy in pediatric renal transplant recipients and to monitor trough levels and side-effects during pre-emptive therapy. Eleven pediatric renal transplant recipients (aged 11.0+/-3.9 years) were included. The diagnosis of CMV infection, based on two positive pp-65 CMV blood antigen tests at 1 week apart, was made at 39+/-12 days post renal transplantation. They received IV GCV at a dose of 5.0+/-0.3 mg/kg per 12 h for 15 days, followed by GCV p.o. at a dose of 46.7+/-8.2 mg/kg per 12 h for 3 months. Pharmacokinetics (PK) were studied at steady state and GCV plasma concentrations were measured by high-performance liquid chromatography. After IV GCV administration, PK parameters were: C(0)=0.84+/-0.66 microg/ml; C(max)=11.77+/-2.82 microg/ml; AUC(0-12 h)=42.29+/-17.57 microg/ml per hour; Cl=0.13+/-0.05 l/h per kg. After p.o. GCV administration, PK parameters were: C(0)=1.08+/-0.68 microg/ml; C(max)=2.70+/-1.07 microg/ml; AUC(0-12 h)=18.97+/ 9.36 microg/ml per hour; Cl/F=2.97+/-1.42 l/h per kg. Bioavailability (F) was 4.9+/-1.2%. Pre-dose concentrations (C(0)) measured under p.o. GCV (n=51) were 1.29+/-0.80 microg/ml (8 C(0) values were below 0.5 microg/ml). Pp-65 CMV blood antigen tests became negative after 16+/-11 days of treatment. GCV was well tolerated. Because of the limited bioavailability, the recommended high doses of p.o. GCV (50 mg/kg per 12 h) were administered and were associated with trough levels over 0.5 microg/ml. In 1 patient who received an erroneously low dosage p.o., CMV resistance to GCV appeared, requiring foscarnet. PMID- 12883978 TI - Serum lipid pattern unifies following renal transplantation in children. AB - Hyperlipidemia is a common problem in solid organ transplant recipients. In this study we evaluated the role of pre-transplant renal replacement therapy on early and late changes of serum lipid levels in children following renal transplantation. In 46 children with chronic renal failure (median age 10.3 years) and 12 children with heart failure (median age 5.0 years), cholesterol and triglycerides were measured before and during follow-up after transplantation. Children with renal failure had significantly higher serum lipids than controls ( n=34, median age 9.2 years) and patients with heart failure. Pre transplantation, cholesterol and triglycerides were significantly lower in the hemodialysis than in the peritoneal dialysis population, whereas conservatively treated children had intermediate levels. After transplantation, serum cholesterol converged towards a mean level of 208 mg/dl and triglyceride levels converged towards a uniform level of 195 mg/dl at 9 months post transplant. The ratio of cholesterol/high-density lipoprotein significantly decreased from 4.7 to 3.8. The pattern of "post-transplant hyperlipidemia" was similar in both renal and cardiac allograft recipients. Hence, the early post-transplant changes of serum lipid pattern are markedly dependent on the mode of pre-transplant renal replacement therapy. Later, serum lipid levels were no longer influenced by prior renal replacement therapy and showed a new pattern of "post-transplant hyperlipidemia" in all children. PMID- 12883979 TI - ACE inhibition modulates transforming growth factor-beta receptors in the young rat. AB - The renin-angiotensin system plays an important role in renal growth and development. Exposure of the neonate to angiotensin converting enzyme (ACE) inhibitors increases mortality and results in growth retardation and abnormal renal development. It has been demonstrated that ACE inhibition in the developing kidney reduces the renal expression of growth factors, which may account for renal growth impairment. This study was designed to investigate the relationship between renal growth impairment and the expression of transforming growth factor beta1 (TGF-beta1), TGF-beta receptor I [TbetaRI, activin-like kinase (ALK)-1 and ALK-5], and TGF-beta receptor II (TbetaRII). Newborn rat pups were treated with enalapril (30 mg/kg per day) or vehicle for 7 days, and kidneys were removed for Western blotting of TGF-beta1, ALK-1, ALK-5, and TbetaRII, and for RT-PCR of ALK 5 and TbetaRII. TGF-beta1, ALK-1, ALK-5, and TbetaRII were also detected by immunohistochemistry. Enalapril treatment resulted in an increased mortality (30.4%) by day 7, and reduced body weight and kidney weight ( P<0.05 versus vehicle). Enalapril decreased renal TGF-beta1, ALK-1, and ALK-5 protein expression ( P<0.05). Also, enalapril decreased ALK-5 mRNA expression ( P<0.05). TbetaRII expression was not changed by enalapril treatment. These results indicate that ACE inhibition in the developing kidney decreases TGF-beta1, ALK-1, and ALK-5 expression, which may account for renal growth impairment. TbetaRII may not be modulated by ACE inhibition in the developing kidney. PMID- 12883980 TI - Vesicoureteric reflux in Kuwaiti children with first febrile urinary tract infection. AB - The prevalence of vesicoureteric reflux (VUR) in children with urinary tract infection (UTI) varies among different racial groups. The purpose of this study was to determine the frequency of VUR and associated renal changes in a group of Arab Kuwaiti children with their first documented febrile UTI and to compare our findings with those reported from other racial groups. One hundred and seventy four children (38 males and 136 females) fulfilled the study criteria and were divided into three age groups (<1 year, 1-5 years, and >5 years). Patients in each group had both micturating cystourethrography (MCUG) and 99m-Tc dimercaptosuccinic acid (DMSA) renal scan after diagnosis. VUR was detected in 39 children (22%). Two-thirds of cases had mild reflux (grade I and II). Females ( n=32) had more reflux than males ( n=7) (24% vs. 18%). Sixty-three patients (36%) had abnormal (DMSA) renal scans (acute pyelonephritis [AP] or renal scars). Of these, 79% were children below 5 years. Abnormal DMSA scans were found in 4 of 38 males (11%) versus 59 of 136 females (43%). Abnormal scans in children with VUR were seen in 1 of 7 males (14%) versus 19 of 32 females (59%). In total, the combination of abnormal scan with VUR occurred in 1 of 38 males (3%) and in 19 of 136 females (14%), whereas abnormal scan without demonstrable VUR was seen in 3 of 38 males (8%) versus 40 of 136 females (29%). Our data showed that the frequency of VUR in Arab Kuwaiti children with febrile UTI is midway between Caucasian and other racial groups. In this study, males had a lower-risk profile than females, the latter having a higher rate of reflux as well as a higher rate of abnormal DMSA scans, irrespective of demonstrable VUR. PMID- 12883981 TI - The use of steroid-sparing agents in steroid-sensitive nephrotic syndrome. AB - Childhood nephrotic syndrome (NS) is frequently characterized by a relapsing course. There is no uniform agreement about the precise stage at which a steroid sparing agent should be introduced to control the disease. In order to evaluate the treatment strategies and outcome of steroid-sensitive NS over the last 2 decades, a retrospective notes review was undertaken in a cohort of children treated at Great Ormond Street Children's Hospital between 1980 and 2000. From a population of 863 children with NS referred, 509 had frequently relapsing or steroid-dependent disease and 261 children received at least one steroid-sparing agent. Cyclophosphamide was the first choice in 178 patients and in 114 no further steroid-sparing agent was needed. Levamisole was prescribed as the first steroid-sparing agent for 65 children and disease control was achieved in 30%. Cyclosporin A was prescribed in 61 children and sustained remission was induced in 69%. It is concluded that cyclophosphamide is a potent agent in inducing sustained remission in steroid-sensitive NS. Levamisole and cyclosporin A have emerged as attractive steroid-sparing agents. Complications and major side effects of treatment are infrequent but occasionally fatal. PMID- 12883982 TI - Morbidity and mortality in children with anemia at initiation of dialysis. AB - Studies in adult dialysis patients have identified anemia as a risk factor for patient morbidity and mortality. Limited comparable outcome data are available from children. We used the database from the North American Pediatric Renal Transplant Cooperative Study (NAPRTCS) to identify patients <18 years of age who initiated dialysis (hemodialysis and peritoneal dialysis) between 1992 and 2001 with no prior history of dialysis or transplantation. A Cox proportional hazards model was used to evaluate the association between anemia (hematocrit <33%) at 30 days post initiation of dialysis and patient mortality. The association between anemia and prolonged hospitalization was also evaluated. Of the 1,942 patients (male 56%), 68% were anemic on day 30. One hundred and seventy-one patient deaths were reported over the 9-year observation period. The multivariate analysis demonstrated anemia to be associated with a 52% higher risk of death (adjusted relative risk 1.52, 95% confidence interval 1.03-2.26, P=0.037). Cardiopulmonary disease was the primary reported cause of death associated with anemia, accounting for 22% of cases. The presence of anemia was also associated with an increased risk for prolonged hospitalization, irrespective of dialysis modality. In conclusion, the presence of anemia 1 month after initiation of dialysis iis associated with an increased risk of prolonged hospitalization and death in pediatric patients. Further studies should ascertain whether anemia per se increases the risk of death or whether it is a marker for other conditions that shorten patient survival, such as chronic inflammation or recurrent infection. PMID- 12883983 TI - Growth hormone aggravates renal abnormalities induced by neonatal enalapril treatment. AB - Lack of neonatal angiotensin II type-1 receptor stimulation produces irreversible abnormalities of renal function and morphology, which can be prevented by simultaneous administration of insulin-like growth factor-I (IGF-I). Given the fact that growth hormone (GH) is the strongest secretagogue for IGF-I, we wanted to explore whether GH could reproduce the effect of IGF-I. Rats were treated from 3 to 13 days of age with the angiotensin-converting enzyme inhibitor enalapril (10 mg/kg/day) and GH (4 mg/kg/day), alone or in combination. Renal gene expression of IGF-I and IGF-binding proteins (IGFBP) was determined during and after treatment. Renal function and morphology were investigated at adult age. In contrast to the beneficial effect of IGF-I, GH treatment in combination with enalapril further deteriorated both renal function and morphology as compared with enalapril treatment alone, demonstrating: reduced glomerular filtration rate, increased tubular dilation and further expansion of the outer medulla. Enalapril decreased medullary expression of IGF-I and increased renal expression of IGFBP-1, changes that were not affected by concomitant GH treatment. These findings indicate that GH and IGF-I have different roles in the renin-angiotensin system-mediated kidney development. PMID- 12883984 TI - Density-dependent foraging behaviors in a parasitoid lead to density-dependent parasitism of its host. AB - Empirical studies of spatial heterogeneity in parasitism by insect parasitoids have focused largely on patterns, while the many possible underlying mechanisms have been little studied in the field. We conducted experimental and observational studies on Tachinomyia similis (Diptera: Tachinidae) attacking western tussock moths (Orgyia vetusta; Lepidoptera: Lymantriidae) on lupine bushes at Bodega Bay, Calif., USA. We examined several foraging behaviors that have been hypothesized to create density-dependent variation in parasitism rates, including spatial aggregation of parasitoids to high host density, mutual interference among searching parasitoids and decelerating functional responses of the parasitoid. At the spatial scale of individual bushes, we detected both aggregation to a high density and a decelerating functional response. The resulting spatial pattern of parasitism was best fit by two models; one included an effect of parasitoid aggregation and the other included an effect of aggregation and a decelerating functional response. Most of the variation in parasitism was not correlated with density of O. vetusta. PMID- 12883985 TI - The field energetics and water fluxes of free-living wombats (Marsupialia: Vombatidae). AB - Wombats are large, fossorial, herbivorous marsupials exhibiting physical and behavioural characteristics indicative of extreme energy conservation. Previous energetics studies have been limited to their basal metabolism under laboratory conditions; little is known of the energetics of free-living wombats. We measured seasonal field metabolic rates (FMR) and water fluxes in the three species of free-living wombat using the doubly labelled water technique, to further investigate the extent of energy conservation in the Vombatidae. Measurements were taken during the wet and dry annual extremes of their characteristically harsh environments, which corresponded to seasonal extremes of food and water availability. Seasonal FMRs for all wombat species were lower than that recorded for other marsupials and well below that predicted for herbivorous mammals. Dry season FMR of Lasiorhinus kreftii was 40% of that predicted for a mammal. Wombats maintained energy balance during the poor season by reducing FMR to about half that of the good season. Water flux rates during the dry season for the arid adapted Lasiorhinus are amongst the lowest recorded for mammals, being only 25% of that predicted for a similarly sized herbivorous mammal. These low water flux rates enable wombats in semi-arid areas to maintain water balance without drinking. Estimated food and nitrogen intake rates were also low. We conclude that the energetically frugal lifestyle of the Vombatidae is amongst the most extreme for mammals. PMID- 12883986 TI - Nitrogen acquisition from inorganic and organic sources by boreal forest plants in the field. AB - A wide range of recent studies have indicated that organic nitrogen may be of great importance to plant nitrogen (N) nutrition. Most of these studies have, however, been conducted in laboratory settings, excluding important factors for actual plant uptake, such as competition, mycorrhizal associations and soil interactions. In order to accurately evaluate the importance of different N compounds to plant N nutrition, field studies are crucial. In this study, we investigated short- as well as long-term plant nitrogen uptake by Deschampsia flexuosa, Picea abies and Vaccinium myrtillus from 15NO3-, 15NH4+ and (U-13C, 15N) arginine, glycine or peptides. Root N uptake was analysed after 6 h and 64 days following injections. Our results show that all three species, irrespective of their type of associated mycorrhiza (arbuscular, ecto- or ericoid, respectively) rapidly acquired similar amounts of N from the entire range of added N sources. After 64 days, P. abies and V. myrtillus had acquired similar amounts of N from all N sources, while for D. flexuosa, the uptake from all N sources except ammonium was significantly lower than that from nitrate. Furthermore, soil analyses indicate that glycine was rapidly decarboxylated after injections, while other organic compounds exhibited slower turnover. In all, these results suggest that a wide range of N compounds may be of importance for the N nutrition of these boreal forest plants, and that the type of mycorrhiza may be of great importance for N scavenging, but less important to the N uptake capacity of plants. PMID- 12883987 TI - Functional correlates of leaf demographic response to gap release in saplings of a shade-tolerant tree, Elateriospermum tapos. AB - For a shade-tolerant SE Asian tropical tree, Elateriospermum tapos (Euphorbiaceae), we studied field-established saplings in gaps and the shaded understory to test the hypothesis that differences in leaf demography and leaf life span under contrasting light regimes should be functionally correlated with architecture, self-shading and nitrogen distribution within the sapling crown. Rates of leaf production and net leaf gain were greater for saplings in gaps than those in the understory. Median leaf life span was approximately 26 months in the gap saplings, while it was estimated to be greater than 38 months in the understory saplings. Consequently, gap saplings had a greater standing leaf number and experienced greater degrees of self-shading than understory saplings. Light availability at individual leaves, estimated by a combination of canopy photos and a three-dimensional architecture model, were negatively correlated with leaf age in gap saplings but not so in understory saplings. Leaf nitrogen content per unit area (Narea) was influenced more by light availability than by leaf age in the gap saplings. In contrast, in understory saplings, Narea was neither correlated with light availability nor with leaf age, and did not decrease significantly before 38 months in leaf age. We conclude that saplings of this shade-tolerant species apparently prolong their leaf life span in the shaded understory through slower rates of leaf production, lower standing number of leaves and lower degrees of self shading than in gap, and that the rate of decline of Narea with leaf age depends on architecture and self-shading regimes that respond to changes in light regimes. PMID- 12883988 TI - Successful lizard eggs in a human-disturbed habitat. AB - As humans extend their influence to an increasingly large portion of the globe, it becomes of both theoretical and practical interest to understand the consequences of our activities on wildlife populations. Norops polylepis is a small anoline lizard native to rainforest of southern Costa Rica. It is also found and known to lay eggs in cattle pastures immediately adjacent to forest. I predicted that N. polylepis eggs would fare poorly in pastures because of the important abiotic and biotic differences with their native forest habitat. I obtained eggs from captive females and estimated an egg's probability of hatching, daily survival rate, and incubation time at various distances along a forest-to-pasture gradient. Contrary to expectation, egg survival rates were higher in pastures than in forest areas. Furthermore, egg incubation times were significantly shorter in pastures than in their native forest. As a result, eggs were at least as likely to hatch in pastures as in forested areas. Thus, pastures may represent a benign or even beneficial habitat for the eggs of some reptile species. High survival of eggs will facilitate range expansion into human-altered habitats such as pastures, but does not guarantee it. Indeed, pastures could represent an ecological trap and a population sink if adults are drawn there (e.g., in search of favorable ovipositioning sites) yet suffer higher mortality than in forests. PMID- 12883989 TI - Are alien plants more competitive than their native conspecifics? A test using Hypericum perforatum L. AB - The evolution of increased competitive ability hypothesis predicts that introduced plants that are long liberated from their natural enemies may lose costly herbivore defense, enabling them to reallocate resources previously spent on defense to traits that increase competitive superiority. We tested this prediction by comparing the competitive ability of native St John's wort (Hypericum perforatum) from Europe with introduced St John's wort from central North America where plants have long grown free of specialist herbivores, and introduced plants from western North America where plants have been subjected to over 57 years of biological control. Plants were grown in a greenhouse with and without competition with Italian ryegrass (Lolium multiflorum). St John's wort from the introduced range were not better interspecific competitors than plants from the native range. The magnitude of the effect of ryegrass on St John's wort was similar for introduced and native genotypes. Furthermore, introduced plants were not uniformly larger than natives; rather, within each region of origin there was a high variability in size between populations. Competition with ryegrass reduced the growth of St John's wort by >90%. In contrast, St John's wort reduced ryegrass growth <10%. These results do not support the contention that plants from the introduced range evolve greater competitive ability in the absence of natural enemies. PMID- 12883990 TI - Water resource partitioning, stem xylem hydraulic properties, and plant water use strategies in a seasonally dry riparian tropical rainforest. AB - This study investigated seasonal variation in the origin of water used by plants in a riparian tropical rainforest community and explored linkages between plant water source, plant xylem hydraulic conductivity and response to the onset of dry conditions. The study focused on five co-dominant canopy species, comprising three tree species ( Doryphora aromatica, Argyrodendron trifoliolatum, Castanospora alphandii) and two climbing palms ( Calamus australis and Calamus caryotoides). Stable isotope ratios of oxygen in water (delta(18)O) from soil, groundwater, stream water and plant xylem measured in the wet season and the subsequent dry season revealed water resource partitioning between species in the dry season. Measurement of stem-area-specific hydraulic conductivity ( K(S)) in the wet season and subsequent dry season showed a significant dry-season loss of K(S) in three of the five species ( Castanospora alphandii, Calamus australis and C. caryotoides) and a decrease in mean K(S) for all species. This loss of hydraulic conductivity was positively correlated with the difference between wet season and dry-season midday leaf water potentials and with leaf carbon isotope discrimination, indicating that plants that were less susceptible to loss of conductivity had greater control over transpiration rate and were more water-use efficient. PMID- 12883991 TI - Defining core habitat of local populations of the gray treefrog (Hyla versicolor) based on choice of oviposition site. AB - Concern over amphibian population declines and loss of terrestrial and aquatic habitat have emphasized the need to define habitat requirements for each stage in a species' life history. The realization that pond-breeding amphibians spend most of their lives in the terrestrial environment suggests the need to protect terrestrial as well as aquatic habitat. Many studies on amphibian populations have focused on emigration from breeding sites to define habitat use; however these studies do not typically elucidate terrestrial activities of adults within the breeding season. We measured colonization rates of artificial pools by gray treefrogs (Hyla versicolor) at multiple distances from natural breeding ponds. We found a non-random distribution of egg deposition among distances, with 95% of eggs deposited within 15 m of the breeding pond. Additionally, we found that the time to first colonization of artificial pools increased with respect to distance. Our results indicate that adult gray treefrogs may travel up to 200 m within a breeding season, and that multiple breeding ponds may be considered part of a single population. We suggest that a minimum core terrestrial habitat of 60 m surrounding breeding sites is appropriate for protection of local populations of gray treefrogs. PMID- 12883992 TI - Localisation of Even-skipped in the mature CNS of the locust, Schistocerca gregaria. AB - Even-skipped (Eve) is a transcriptional repressor found within the nuclei of a subset of neurons in the embryonic central nervous system of insects. It is necessary for neuronal pathfinding and promotes adhesion of developing motor axons to the intersegmental nerve. Here we show by immunohistochemical staining that Eve is also expressed in the nuclei of a small set of neurons in the thoracic nervous system of larval and adult locusts, suggesting a postembryonic role in transcriptional control. Thirty-two to 39 Eve-positive neurons in four distinct subsets were found in the mesothoracic ganglion of fourth instar larvae and in adults: an anterior, bilaterally paired group of 2-4 neurons; a posterior, bilaterally paired group of 12-15 neurons; 1-2 neurons in a midline position, at the centre of the ganglion; and 1-2 neurons near the midline in a posterior position. These subsets did not change in position or number of neurons within them from fourth instars through to adults, 2 and 35 days post-final moult. The metathoracic ganglion had approximately 48 Eve-positive neurons in four subsets: an anterior, bilaterally paired group of 3-4 neurons; a posterior, bilaterally paired group of 8-10 neurons; a diffuse group of 19 neurons at the midline and a group of 4 neurons in a midline, posterior position. These results suggest that Eve demonstrates both phasic and persistent expression throughout the insect's lifetime and that changes in expression are segment-specific. We suggest Eve helps maintain neural phenotype in the adult CNS, possibly by regulating the expression of cell adhesion molecules. PMID- 12883993 TI - Endothelial-pericyte interactions in angiogenesis. AB - It takes two to make blood vessels-endothelial cells and pericytes. While the endothelial cells are the better characterized of the two, pericytes are now coming into focus as important regulators of angiogenesis and blood vessel function, and as potential drug targets. However, pericytes are still surrounded by much controversy. They are difficult to define, they constitute a heterogeneous population of cells, and their ontogeny is not well understood. They are plastic and have the capacity to differentiate into other mesenchymal cell types, such as smooth muscle cells, fibroblasts and osteoblasts. Recent interest in pericytes also stems from their potential involvement in diseases such as diabetic microangiopathy, tissue fibrosis, cancer, atherosclerosis and Alzheimer's disease. The present review focuses on the role of pericytes in physiological angiogenesis. The currently favored view states that the initial endothelial tubes form without pericyte contact, and that subsequent acquisition of pericyte coverage leads to vessel remodeling, maturation and stabilization. Improved means of identifying and visualizing pericytes now challenge this view and show that high numbers of pericytes invest in actively sprouting and remodeling vessels. Genetic data demonstrate the critical importance of pericytes for vascular morphogenesis and function, and imply specific roles for the cell type in various aspects of angiogenesis. PMID- 12883994 TI - Prox1, master regulator of the lymphatic vasculature phenotype. AB - In contrast to the extensive molecular and functional characterization of blood vascular endothelium, little is known about the mechanisms that control the formation and lineage-specific differentiation and function of lymphatic vessels. The homeobox gene Prox1, the vertebrate homologue of the Drosophila prospero gene, has been recently identified to be required for the induction of lymphatic vascular development from preexisting embryonic veins, and studies in Prox1 deficient mice have confirmed Florence Sabin's original hypothesis about the origin of the lymphatic vascular system from embryonic veins. The recent establishment of cell culture models for the selective propagation of blood vascular and lymphatic endothelial cells, together with the findings that these cells maintain their lineage-specific differentiation in vitro, has led to the discovery that Prox1 expression is sufficient to induce a lymphatic phenotype in blood vascular endothelium. Ectopic expression of Prox1 downregulated blood vascular-associated genes and also upregulated some of the known lymphatic endothelial cell markers. Together, these studies suggest that the blood vascular phenotype represents the default endothelial differentiation and they identify an essential role of Prox1 in the program specifying lymphatic endothelial cell fate. PMID- 12883995 TI - Lymphangiogenesis and tumor metastasis. AB - The lymphatic system transports interstitial fluid and macromolecules from tissues back to the blood circulation, and plays an important role in the immune response by directing the traffic of lymphocytes and antigen-presenting cells. The lymphatic system also constitutes one of the most important pathways of tumor dissemination. In many human cancers, increased expression of vascular endothelial growth factor-C (VEGF-C) is correlated with regional lymph node metastases. Experimental studies using transgenic mice overexpressing VEGF-C or xenotransplantation of VEGF-C-expressing tumor cells into immunodeficient mice have demonstrated a role for VEGF-C in tumor lymphangiogenesis and the subsequent formation of lymph node metastases. However, there is at present little evidence for lymphangiogenesis in human tumors and the relative importance of preexisting vs. newly formed lymphatics for metastasis in humans remains to be determined. Nonetheless, the striking correlation between the levels of VEGF-C in primary human tumors and lymph node metastases predicts its importance in cancer spread. Aside from promoting lymphangiogenesis, VEGF-C may also activate lymphatics to promote tumor cell chemotaxis, lymphatic intravasation and hence tumor cell dissemination. PMID- 12883996 TI - Expression of the small conductance Ca2+-activated K+ channel, SK3, in the olfactory ensheathing glial cells of rat brain. AB - Olfactory sensory neurons are wrapped by ensheathing glial cells in the olfactory nerve layer (ONL). Neither functional roles nor electrical properties of ensheathing glial cells have been, as yet, fully clarified. Four subunits (SK1-4) of small conductance Ca2+-activated K+ (SK) channels have been cloned. In the present study, immunohistochemical analyses showed that SK3 channels are expressed in ensheathing glial cells in the rat olfactory bulb, in addition to neuronal cells in other regions. Western blotting analysis demonstrated that SK3 was predominantly expressed in the olfactory bulb, thalamus, moderately in the hippocampus and cerebellum and modestly in the cerebral cortex of the rat brain. SK3 immunoreactivity was detected in the ONL of the olfactory bulb, neural cell body and fibers of the substantia nigra and hypothalamus. SK3 immunoreactivity was quite intense in the outer (superficial) part of the ONL. SK3-immunoreactive structures were overlapped with glial fibrillary acidic protein (GFAP), but not with vimentin, markers for glial cells and olfactory sensory axons, respectively. Immunoelectron microscopy showed that SK3 immunoreactivity was localized in thin processes that enfolded fascicles of immunonegative olfactory nerve axons. These results indicate that SK3 is expressed specifically in the olfactory ensheathing glial cells in olfactory regions. PMID- 12883997 TI - ATP-evoked increase in intracellular calcium via the P2Y receptor in proliferating bovine trophoblast cells. AB - Bovine trophoblasts actively proliferate to elongate blastocysts before implantation. The trophoblast at this stage secretes cytokines and starts to differentiate into an endocrine cell (binucleate cell) for successful pregnancy. Intracellular calcium ([Ca2+]i) may act as a second messenger in the trophoblast response. In this study, we investigated [Ca2+]i signals in a bovine trophoblast cell line (BT-1) using fura-2 fluorescence. We found that an application of ATP (> or =1 microM) induced a transient increase in [Ca2+]i in BT-1 cells. The ATP induced increase was not affected by the removal of extracellular Ca2+, but was suppressed by suramin (100 microM), an antagonist of P2 receptors. Pretreatment with pertussis toxin (0.1 or 1 microg/ml) partially inhibited the response to ATP. The order of potency to increase [Ca2+]i was ATP=UTP>ADP. ATP-induced [Ca2+]i responses preferentially occurred in cells at the periphery of the colony. The reduced responses at the center of the colony were associated with an increase in cell density and decrease in bromodeoxyuridine incorporation. These results indicated that ATP stimulated P2Y receptors coupled to pertussis toxin sensitive and -insensitive G proteins, leading to an increase in [Ca2+]i as a result of release of Ca2+ from intracellular stores in BT-1 cells. The occurrence of ATP-induced [Ca2+]i signals depended on the cell confluence and reflected the high proliferative activity of the trophoblast cell population. PMID- 12883998 TI - STRO-1, HOP-26 (CD63), CD49a and SB-10 (CD166) as markers of primitive human marrow stromal cells and their more differentiated progeny: a comparative investigation in vitro. AB - There is widespread interest in the use of bone marrow stromal cells (BMSC) for tissue reconstruction and repair and for gene therapy. BMSC represent the differentiated progeny of CFU-F, which however comprise a developmentally heterogeneous population as is reflected in the cellular heterogeneity of the cell populations to which they give rise. We have compared the efficacy of monoclonal antibodies recognising a series of stromal antigens, viz. STRO-1, HOP 26, CD49a and SB-10/CD166, as tools for the enrichment of CFU-F prior to culture and as developmental markers for culture-expanded BMSC. In freshly isolated bone marrow mononuclear cells (BMMNC), the proportion of antigen-positive cells was 27%, 46%, 5% and 19% for STRO-1, HOP-26, CD49a and CD166, respectively. All CD49a(+) cells co-expressed STRO-1. The degree of CFU-F enrichment obtained with anti-CD49a (approximately 18-fold) by a one-pass immunoselection strategy was significantly greater than that of all other antibodies tested. BMSC expressed higher levels of all antigens investigated (except for HOP-26) compared with BMMNC. Expression of STRO-1 and CD49a remained restricted to a subset of BMSC, whereas all BMSC were SB-10/CD166 positive. Treatment with dexamethasone (10 nM), which promotes the differentiation and further maturation of cells of the osteogenic lineage in this cell culture system, increased the expression of CD49a and HOP-26. The CD49a(+) and HOP-26(+) fractions of BMSC were further subdivided by dual-labelling with anti-STRO-1 and B4-78 (an antibody recognising the B/L/K isoform of the enzyme alkaline phosphatase), respectively. By using a variety of criteria, the HOP-26 antigen was identified as CD63, a member of the tetraspanin family of proteins thought to modulate integrin compartmentalisation and signalling. PMID- 12883999 TI - A novel method for automatic genotyping of microsatellite markers based on parametric pattern recognition. AB - Genetic mapping of loci affecting complex phenotypes in human and other organisms is presently being conducted on a very large scale, using either microsatellite or single nucleotide polymorphism (SNP) markers and by partly automated methods. A critical step in this process is the conversion of the instrument output into genotypes, both a time-consuming and error prone procedure. Errors made during this calling of genotypes will dramatically reduce the ability to map the location of loci underlying a phenotype. Accurate methods for automatic genotype calling are therefore important. Here, we describe novel algorithms for automatic calling of microsatellite genotypes using parametric pattern recognition. The analysis of microsatellite data is complicated both by the occurrence of stutter bands, which arise from Taq polymerase misreading the number of repeats, and additional bands derived form the non-template dependent addition of a nucleotide to the 3' end of the PCR products. These problems, together with the fact that the lengths of two alleles in a heterozygous individual may differ by only two nucleotides, complicate the development of an automated process. The novel algorithms markedly reduce the need for manual editing and the frequency of miscalls, and compares very favourably with commercially available software for automatic microsatellite genotyping. PMID- 12884000 TI - Allelic variation at alcohol metabolism genes ( ADH1B, ADH1C, ALDH2) and alcohol dependence in an American Indian population. AB - Enzymes encoded by two gene families, alcohol dehydrogenase ( ADH) and aldehyde dehydrogenase ( ALDH), mediate alcohol metabolism in humans. Allelic variants have been identified that alter metabolic rates and influence risk for alcoholism. Specifically, ADH1B*47His (previously ADH2-2) and ALDH2-2 have been shown to confer protection against alcoholism, presumably through accumulation of acetaldehyde in the blood and a resultant 'flushing response' to alcohol consumption. In the current study, variants at ADH1B (previously ADH2), ADH1C (previously ADH3), and ALDH2 were assayed in DNA extracts from participants belonging to a Southwest American Indian tribe ( n=490) with a high prevalence of alcoholism. Each subject underwent a clinical interview for diagnosis of alcohol dependence, as well as evaluation of intermediate phenotypes such as binge drinking and flushing response to alcohol consumption. Detailed haplotypes were constructed and tested against alcohol dependence and related intermediate phenotypes using both association and linkage analysis. ADH and ALDH variants were also assayed in three Asian and one African population (no clinical data) in order to provide an evolutionary context for the haplotype data. Both linkage and association analysis identified several ADH1C alleles and a neighboring microsatellite marker that affected risk of alcohol dependence and were also related to binge drinking. These data strengthen the support for ADH as a candidate locus for alcohol dependence and suggest further productive study. PMID- 12884002 TI - The cnm locus, a canine homologue of human autosomal forms of centronuclear myopathy, maps to chromosome 2. AB - Myotubular/centronuclear myopathies are a nosological group of hereditary disorders characterised by severe architectural and metabolic remodelling of skeletal muscle fibres. In most myofibres, nuclei are found at an abnormal central position within a halo devoid of myofibrillar proteins. The X-linked form (myotubular myopathy) is the most prevalent and severe form in human, leading to death during early postnatal life. Maturation of fibres is not completed and fibres resemble myotubes. Linkage analysis in human has helped to identify MTM1 as the morbid gene. MTM1 encodes myotubularin, a dual protein phosphatase. In families in which myotubular myopathy segregates, detected mutations in MTM1 abolish the specific phosphatase activity targeting the second messenger phosphatidylinositol 3-phosphate. Autosomal forms (centronuclear) have a later onset and are often compatible with life. At birth, fibres are normally constituted but progressively follow remodelling with a secondary centralisation of nuclei. Their prevalence is low; hence, no linkage data can be performed and no molecular aetiology is known. In the Labrador Retriever, a spontaneous disorder strikingly mimics the clinical evolution of the human centronuclear myopathy. We have established a canine pedigree and show that the disorder segregates as an autosomal recessive trait in that pedigree. We have further mapped the dog locus to a region on chromosome 2 that is orthologous to human chromosome 10p. To date, no human MTM1 gene member has been mapped to this genetic region. This report thus describes the first spontaneous mammalian model of centronuclear myopathy and defines a new locus for this group of diseases. PMID- 12884003 TI - Relationship between serum HDL-C levels and common genetic variants of the endothelial lipase gene in Japanese school-aged children. AB - Endothelial lipase (EL) is a new member of the triglyceride lipase family, the genes of which play a central role in dietary fat absorption, energy homeostasis, and plasma lipoprotein metabolism. One physiologic role of EL is thought to be hydrolysis of high-density lipoprotein (HDL) phospholipid, although the precise function of endothelial lipase has yet to be fully clarified. Furthermore, genetic variation in EL has been suggested to influence serum HDL-C levels. In the present study, we detected two common single nucleotide polymorphisms in the EL gene associated with serum HDL cholesterol levels in healthy school-aged children. Our data support the hypothesis that variations in the EL gene are one of the genetic determinants of serum HDL-C levels. PMID- 12884004 TI - Exon skipping caused by an intronic insertion of a young Alu Yb9 element leads to severe hemophilia A. AB - Short interspersed elements, such as Alu elements, have propagated to more than one million copies in the human genome. They affect the genome in several ways, caused by retrotransposition, recombination between elements, gene conversion, and alterations in gene expression. These events, including novel insertions into active genes, have been associated with a number of human disorders. Hemophilia A is an X-linked severe bleeding disorder and is caused by mutations in the Factor VIII gene. The spectrum of mutations includes point mutations, rearrangements, insertions, and deletions. Recently, an Alu retrotransposition event in a coding exon has been reported in a family with a severe form of hemophilia A. This was the first report of an Alu insertion in the Factor VIII gene. Here, we report a second Alu insertion event that lies in an intron of the same gene that causes exon skipping and the complete disruption of gene expression. PMID- 12884005 TI - Evaluation of bidirectional transfer of plasma DNA through placenta. AB - To clarify the origin of cell-free fetal DNA in maternal plasma, we analyzed bidirectional transfer of plasma DNA between fetus and mother. We analyzed maternal and fetal plasma DNA obtained from 15 pregnant women at the time of Cesarean section. The subjects were five patients with preeclampsia and 10 gestational-age-matched normal controls. DNA was extracted from 1.5-ml plasma samples and the cellular fraction of maternal and umbilical blood. Seven polymorphic marker genes were analyzed. The relative concentration of fetal DNA in maternal plasma and maternal DNA in cord blood were evaluated. The relative concentration of maternal DNA in fetal circulation (median, 0.9%; range, 0.2 8.4%) was significantly lower than that of fetal DNA in maternal blood (14.3%, 2.3-64%), with P=0.007. The relative concentration of maternal DNA in fetal blood was not affected by preeclampsia. These findings indicate that cell-free DNA is unequally transferred through the placenta. The structural characteristics of the placenta suggest that the majority of cell-free fetal DNA in maternal plasma is derived from villous trophoblasts. PMID- 12884007 TI - An integrated physical and genetic map of the nematode Pristionchus pacificus. AB - The free-living nematode Pristionchus pacificus is one of several species that have recently been developed as a satellite system for comparative functional studies in evolutionary developmental biology. Comparisons of developmental processes between P. pacificus and the well established model organism Caenorhabditis elegans at the cellular and genetic levels provide detailed insight into the molecular changes that shape evolutionary transitions. To facilitate genetic analysis and cloning of mutations in P. pacificus, we previously generated a BAC-based genetic linkage map for this organism. Here, we describe the construction of a physical map of the P. pacificus genome based on AFLP fingerprint analysis of 7747 BAC clones. Most of the SSCP markers used to generate the genetic linkage map were derived from BAC ends, so that the physical genome map and the genetic map can be integrated. The contigs that make up the physical map are evenly distributed over the genetic linkage map and no clustering is observed, indicating that the physical map provides a valid representation of the P. pacificus genome. The integrated genome map thus provides a framework for positional cloning and the study of genome evolution in nematodes. PMID- 12884008 TI - Genome-wide survey of mRNA half-lives in Bacillus subtilis identifies extremely stable mRNAs. AB - We have used DNA microarrays to survey rates of mRNA decay on a genomic scale in early stationary-phase cultures of Bacillus subtilis. The decay rates for mRNAs corresponding to about 1500 genes could be estimated. About 80% of these mRNAs had a half-life of less than 7 min. More than 30 mRNAs, including both mono- and polycistronic transcripts, were found to be extremely stable, i.e. to have a half life of > or =15 min. Only two such transcripts were known previously in B. subtilis. The results provide the first overview of mRNA decay rates in a gram positive bacterium and help to identify polycistronic operons. We could find no obvious correlation between the stability of an mRNA and the function of the encoded protein. We have also not found any general features in the 5' regions of mRNAs that distinguish stable from unstable transcripts. The identified set of extremely stable mRNAs may be useful in the construction of stable recombinant genes for the overproduction of biomolecules in Bacillus species. PMID- 12884009 TI - Nucleotide binding site/leucine-rich repeats, Pto-like and receptor-like kinases related to disease resistance in grapevine. AB - Nucleotide Binding Site/Leucine-Rich Repeat (NBS-LRR) and Serine/Threonine Kinase (STK) genes are two of the known classes of resistance (R-) genes in plants, and occur in large multigene families. Systematic identification of genes for NBS LRRs and STKs provides a means of access to genomic regions that may be involved in disease resistance. Here we present a picture of these two families of R-gene analogs (RGAs) in grape with the aim of developing a set of resistance-related sequence-tagged-site (STS) markers. One hundred and three NBS-LRR sequences were isolated. They included members of the CC (coiled-coil) and TIR (Toll-interleukin receptor) sub-classes. A comparative analysis with other angiosperm NBSs is provided. Fifty-three genes for receptor-like kinases (RLKs) with serine/threonine specificity were identified. RLK sequences formed a putative monophyletic group within the kinase superfamily. They were similar to both cytoplasmic RLKs, such as Pto, and RLKs with LRR, S-locus, lectin-like and thaumatin-like extracellular binding-domains. The latter resembled the products of the R-related genes Xa21, FLS2, Rlk10, SFR2, and PR5K. Forty-five reference RGAs were converted into STSs by using appropriately designed specific primers. RGA-STSs were present in diverse grape genotypes, and >85% of the primers were capable of amplifying the STSs across the taxa Vitis and Muscadinia. DNA sequence polymorphism among these RGAs was assessed by SSCP (single-strand conformation polymorphism) analysis in over 20 Vitis spp. Finally, 45 universal primers for grape RGAs are proposed that should permit tagging of R-related regions in any grape genome. PMID- 12884010 TI - Molecular analysis of two cytochrome P450 monooxygenase genes required for paxilline biosynthesis in Penicillium paxilli, and effects of paxilline intermediates on mammalian maxi-K ion channels. AB - The gene cluster required for paxilline biosynthesis in Penicillium paxilli contains two cytochrome P450 monooxygenase genes, paxP and paxQ. The primary sequences of both proteins are very similar to those of proposed cytochrome P450 monooxygenases from other filamentous fungi, and contain several conserved motifs, including that for a haem-binding site. Alignment of these sequences with mammalian and bacterial P450 enzymes of known 3-D structure predicts that there is also considerable conservation at the level of secondary structure. Deletion of paxP and paxQ results in mutant strains that accumulate paspaline and 13 desoxypaxilline, respectively. These results confirm that paxP and paxQ are essential for paxilline biosynthesis and that paspaline and 13-desoxypaxilline are the most likely substrates for the corresponding enzymes. Chemical complementation of paxilline biosynthesis in paxG (geranygeranyl diphosphate synthase) and paxP, but not paxQ, mutants by the external addition of 13 desoxypaxilline confirms that PaxG and PaxP precede PaxQ, and are functionally part of the same biosynthetic pathway. A pathway for the biosynthesis of paxilline is proposed on the basis of these and earlier results. Electrophysiological experiments demonstrated that 13-desoxypaxilline is a weak inhibitor of mammalian maxi-K channels (Ki=730 nM) compared to paxilline (Ki=30 nM), indicating that the C-13 OH group of paxilline is crucial for the biological activity of this tremorgenic mycotoxin. Paspaline is essentially inactive as a channel blocker, causing only slight inhibition at concentrations up to 1 microM. PMID- 12884011 TI - Observation on the schizont stage of an unidentified Theileria sp. in experimentally infected sheep. AB - The schizont stage of an unidentified Theileria sp. infective for small ruminants was observed. Intact sheep were infected with adult Haemaphysalis qinghaiensis collected from fields where ovine theileriosis was prevalent. When the infested sheep developed theileriosis, tissue and organ smears were prepared. Theileria schizonts were demonstrated in liver, spleen, lung, kidney, lymph node and peripheral blood. Most of the schizonts were found outside the host cells, which was probably a smear artifact. This study adds our knowledge that lung and kidney can be parasitized by schizonts of Theileria sp. PMID- 12884012 TI - Prevalence and risk factors for enterobiasis among preschool children in a metropolitan city in Korea. AB - The infection rate of Enterobius vermicularis was investigated in 1,191 preschool children in 25 daycare centers in Seoul, Korea by cellotape anal swab from July to August 1999. Both the directors of the daycare centers and the children's parents were asked to complete questionnaires that inquired about the potential risk factors involved. The overall egg positive rate for E. vermicularis was 9.5%, and the prevalence in the daycare centers ranged from 0 to 31.1%. Children aged 6-7 years showed a significantly higher egg positive rate than younger children, but the infection rate was similar for boys and girls. The socioeconomic status of the family and personal hygiene were not associated with enterobiasis, but anthelmintic medication significantly reduced the infection rate. The daycare centers in residential areas showed significantly lower egg positive rates than those situated near traditional markets. The environment of daycare centers is an important factor in enterobiasis, and the mass screening and treatment of children at high risk is recommended. PMID- 12884013 TI - Selenium supplementation at low doses contributes to the decrease in heart damage in experimental Trypanosoma cruzi infection. AB - Chagasic patients with cardiomyopathy have low levels of selenium (Se), a fundamental trace element. We evaluated the effect of supplementing infected mice with Se (0.25-16 ppm). Supplementation with 0.25 or 1 ppm Se led to parasitaemia and survival curves similar to those of the control group. Mice treated with 4-16 ppm showed a dose-dependent decrease of parasitaemia, significant for the highest concentration. This was probably due to a direct effect on the parasites, which were lysed after in vitro incubation with Se. Survival rates did not change significantly; however, heart damage was reduced in infected mice supplemented with 4 ppm Se, as indicated by a lower cardiac isoform of creatine kinase levels. Our results imply that Se supplementation does not lead to a general protection during infection, but may help protect the heart from inflammatory damage. The effect of Se supplementation in the course of T. cruzi infection depends on the host-parasite pair employed. PMID- 12884014 TI - Genetic and phylogeographic structure of populations of Pulex simulans (Siphonaptera) in Peru inferred from two genes ( CytB and CoII). AB - In this paper we discuss the potential usefulness of determining the phylogeographic and phylogenetic patterns of a vector for understanding the spread of pathogens or insecticide resistance. We do so using the example of Pulex simulans in Peru. Six populations from six different localities were investigated. Mitochondrial DNA sequences were obtained and branching patterns were inferred using phylogenetic reconstruction methods and nested clade analyses. Ten different haplotypes were discovered. Phylogenetic analysis revealed P. simulans in Peru as a monophyletic group, containing clades that were generally not geographically correlated. The data suggest that P. simulans is not a single genetic entity but rather that this species shows a high degree of intraspecific variation. Restricted gene flow with long distance dispersal coupled with range expansion and long distance colonization are likely to have contributed to the observed patterns of variation. PMID- 12884015 TI - Intestinal helminths of a landlocked ringed seal (Phoca hispida saimensis) population in eastern Finland. AB - A small, landlocked, endangered ringed seal (Phoca hispida saimensis) population lives as a postglacial relict in Lake Saimaa in eastern Finland. In this study, the intestinal metazoans were examined from a total of 61 Saimaa seals found dead from 1981 to 2001. The helminth fauna was very depauperate. Only one acanthocephalan species, Corynosoma magdaleni, has been able to survive during isolation in the freshwater environment. In addition, only two cestode species were found: Diphyllobothrium ditretum and Schistocephalus sp. However, neither of these larvae developed in the ringed seals. As the newborn pups of Saimaa seals are nursed for about 2 months only one of them was infected by C. magdaleni. At 2 5 months the pups harboured worms; however, the mean intensity did not rise distinctively until the seals were adults. C. magdaleni infection increased with increasing age and the abundances were significantly different between the age groups. There was no correlation between the number of parasites and the adult seal body weight; however, seals weighing 36 kg or more carried the majority of the worms. The distribution of the parasites was aggregated; six seals carried 65% (n=1,182) of all worms (total n=1,809) and 26 seals were uninfected. Variation in the number of worms in individual seals may indicate differences in feeding behaviour and in the diet of seals, and/or variation in the immune status of the seals. PMID- 12884016 TI - Detection of glial fibrillary acidic protein and neurofilaments in the cerebrospinal fluid of patients with neurocysticercosis. AB - Neurocysticercosis (NCC) is an infection caused by Taenia solium larval metacestodes in the central nervous system. The glial fibrillary acidic protein (GFAP) and neurofilaments (NFs) can be used as markers of glial and neuronal damage, respectively. We studied the GFAP and NFs of 68, 160 and 200 kDa in the cerebrospinal fluid (CSF) of patients with NCC by Western blotting. Our results showed that patients with NCC had significantly elevated GFAP levels in the CSF compared with the control, whereas NFs of 68, 160 and 200 kDa were not detected in the CFS of NCC patients. We concluded that GFAP could be used as a marker of glial damage in the CFS of NCC patients. PMID- 12884017 TI - The monocyte locomotion inhibitory factor produced by Entamoeba histolytica inhibits induced nitric oxide production in human leukocytes. AB - The monocyte locomotion inhibitory factor, an anti-inflammatory pentapeptide produced by Entamoeba histolytica, inhibits the in vitro production of nitric oxide induced by cytokines (INF-gamma, TNF-alpha) or PMA in human leukocytes. This can be added to the other previously reported functional effects of this factor, such as the inhibition of monocyte locomotion and the synthesis of reactive oxygen intermediates in both monocytes and neutrophils. The decreased nitric oxide production may interfere with the killing of amebas by neutrophils in the early invasive stages of amebiasis, when oxidative mechanisms are used [reactive oxygen and nitrogen intermediates either individually or synergistically via peroxynitrite (ONOO(-))], and in the advanced stages, when both non-oxidative and oxidative (including nitric oxide) mechanisms are employed by macrophages. Diminished nitric oxide production by leukocytes may also contribute to the paucity of late inflammatory components in amebic abscess of the liver and other amebic lesions. PMID- 12884018 TI - Ion regulation in the different life stages of Trypanosoma cruzi. AB - Different ion and pH regulation mechanisms have been detected in the three main life stages of Trypanosoma cruzi: epimastigote, metacyclic trypomastigote and amastigote. Treatment with amiloride showed that the Na(+)/H(+) exchanger participated in all three forms. The Na(+)/K(+) ATPase exchanger appeared to be more active in the epimastigote than in the other forms. V-H(+)-ATPase inhibitors revealed the activity of this regulatory mechanism in the amastigote and epimastigote forms, while treatment with oligomycin only affected the amastigotes. The HCO(-)(3)/Cl(-) exchanger was found in all stages as well as in the intracellular pH-regulatory mechanism after abrupt basification. We deduce that ion regulation in T. cruzi is a complex process and depends upon the precise stage of the cell cycle of the parasite. It would seem to be an important mechanism, allowing the parasite to adapt to the changing environmental conditions within which it develops. PMID- 12884019 TI - Taenia solium metacestode antigens which are protective for pigs induce Th1/Th2 mixed responses in mice. AB - The purpose of this study was to determine the Th1 and Th2 cytokine responses induced by Taenia solium metacestode antigens in mice and correlate them with the immune responses elicited in vivo. To assess this aim, mice were inoculated with metacestode antigens. RNA was obtained from spleen cells of immunized or control mice incubated with metacestode antigens and used to determine the cytokine profile. Peripheral blood eosinophilia was measured daily in each mouse and specific serum antibody levels were determined. Results showed that metacestode antigens induce the synthesis of IL-4, IL-5 and IFN-gamma mRNAs in spleen cells. They also induced peripheral blood eosinophilia and elicited specific IgE and IgG antibodies, especially IgG1. Three antigens were recognized by all IgG subclasses and by IgE (104, 88 and 7 kDa), and a 57-kDa protein was recognized by IgG1, IgG2a, IgG2b, and IgE. IgG1 and IgG2b recognized 52, 30 and 20 kDa antigens. Immune responses elicited in vivo and the cytokine profile showed good correlation. PMID- 12884020 TI - Cuticular Architecture of Hassalstrongylus epsilon (Nematoda: Trichostrongyloidea). AB - Hassalstrongylus epsilon is a small nematode, whose adult forms are found among the intestinal microvilli of the water rat Nectomys squamipes, Brants 1827 (Rodentia: Muridae). The external appearance of the cuticle, which presents transversal striations and longitudinal ridges, is described using scanning electron microscopy. Transmission electron microscopy of thin sections and replicas of quick-frozen, freeze-fractured, deep-etched and rotatory shadowed samples showed the presence in the cuticle of struts that arise from the fluid median layer, extending outward to the epicuticle. The cuticle showed the presence of five layers: epicuticle, cortical, fibril-rich, fluid median and fibrous. The cuticle layers were made of an assemblage of fibers that create compartments, which were larger in the fluid region than in the fibril-rich median layer. PMID- 12884021 TI - Analysis of the gene expression profile of Schistosoma japonicum cercariae by a strategy based on expressed sequence tags. AB - We present an analysis of the expression profile of Schistosoma japonicum cercariae by a strategy based on expressed sequence tags (ESTs). A cDNA library from S. japoniucm cercariae was constructed and was used to generate ESTs. In total, 201 clones randomly selected from the library were sequenced; 136 ESTs were successfully obtained and sent to the BLAST server for homology searching. Among the 136 ESTs, 85 (62.50%) did not match any protein or gene sequence published in the BLAST databases; these comprised 75 (55.15%) ESTs matched (or partly matched) S. japonicum and Schistosoma mansoni ESTs, 4 (2.94%) matched human ESTs and 6 (4.41%) that did not match any sequence. Fifty-one (37.50%) ESTs were identified by the BLAST server; these consisted of 8 (5.88%), 9 (6.62%) and 34 (25.00%) that showed high homology with genes or proteins reported from S. japonicum, S. mansoni and other organisms, respectively. These identified ESTs can be grouped into nine categories: transporters (1.96%), secretory proteins (1.96%), kinases (3.92%), proteases (5.88%), structural and cytoskeletal proteins (13.73%), metabolism-related proteins (9.80%), regulatory and signaling proteins (11.76%), transcription and translation machinery (25.50%), and others (25.50%). Several interesting new genes cloned from this cDNA library are discussed here. These findings will be valuable for the understanding of the biology of this parasite. PMID- 12884022 TI - Construction of an EGF receptor-mediated histone H1(0)-based gene delivery system. AB - PURPOSE: To construct an EGF receptor (EGF-R)-mediated histone H1(0)-based gene delivery system for gene therapy. METHODS: A recombinant DNA containing histone H1(0), EGF-R ligand, and endosomalytic domains was constructed in a prokaryotic vector and expressed in E. coli. Expression of the beta-galactosidase (beta-gal) gene in the tumor cells and tissues was observed after transduction of the beta gal gene packaged by purified fusion proteins in vitro and in vivo. RESULTS: As an extension of the research on previously reported chemically synthetic composite polypeptide gene delivery systems, this genetically engineered polypeptide has proved to be capable of targeting the beta-galactosidase (beta gal) gene into EGF-R-positive cancer cells both in vitro and in vivo. We also studied the time course of beta-gal gene expression in tumor tissues delivered in vivo by this polypeptide vector. At 24 h after administration, expression of the beta-galactosidase gene in tumor reached peak levels. The dosage optimization of administered polyplex was also investigated. The optimal dose of polyplex per mouse was 1 microg DNA packaged by 3 microg of composite polypeptide. CONCLUSIONS: The genetically engineered polypeptide based on histone H1(0) is a promising gene delivery system targeting EGF-R. PMID- 12884023 TI - Correlation between GP-170 expression, prognosis, and chemoresistance of superficial bladder carcinoma. AB - PURPOSE: To study GP-170 in superficial bladder cancer at initial diagnosis and at recurrence and to evaluate if intravesical chemoprophylaxis modifies the expression of GP-170 in tumor recurrences. MATERIALS AND METHODS: GP-170 was retrospectively assessed in 160 patients affected by primary superficial transitional cell carcinoma of the bladder and followed for up to 10 years. Eighty-four patients (52.5%) recurred after transurethral resection (TUR). Adjuvant intravesical chemotherapy after TUR was adopted in 52 patients. The correlations between GP-170 and G-grade, T-category, risk of recurrence and of progression, and adoption of adjuvant intravesical chemotherapy were investigated. The correlations between variations in grade and stage at recurrence and modifications in GP-170 expression were also studied. RESULTS: No significant correlation between GP-170 expression and G-grade and T-category was found. A significant correlation was detected between GP-170 expression and recurrence ( P=0.0383). It showed a biphasic pattern, i.e., tumors that did not express GP-170 had a higher recurrence rate, but high GP-170 levels were also associated with an increasing risk of recurrence. Intravesical chemotherapy did not induce significative variations in GP-170 expression. No correlation was found between progression and GP-170. CONCLUSION: GP-170 seems to be an independent prognostic factor for recurrence in superficial bladder tumors. A negative GP-170 pattern and high levels of GP-170 are associated with an increasing risk of recurrence but have no impact upon progression. In our experience, GP-170 is neither induced nor modified by intravesical chemotherapy, although it might represent a factor of chemoresistance when strongly expressed. PMID- 12884024 TI - Analysis of cyclooxygenase-2 expression in human breast cancer: high throughput tissue microarray analysis. AB - PURPOSE: The objective of this study was to evaluate breast carcinomas for the expression of cyclooxygenase-2 (Cox-2) using a tissue microarray (TMA) and to determine its clinical and prognostic relevance. METHODS: We analyzed Cox-2 expression in 600 samples from 200 breast carcinomas immunohistochemically performing TMA technology and semiquantitative analysis. Results were correlated with various clinicopathological variables and follow-up data. Expression of estrogen receptor, progesterone receptor, Ki-67, and Her-2/neu-oncogene was analyzed and correlated with Cox-2 status. RESULTS: We observed a moderate or strong cytoplasmic staining for Cox-2 in 78 (40.6%) of breast carcinomas. Increased Cox-2 expression corresponded to higher pT stage ( P=0.038), amplification of Her-2/neu ( P=0.032), lymphovascular invasion ( P=0.006), a high MIB-1 labeling index (LI) ( P<0.001), and histological grading ( P=0.013). We also observed an inverse relationship between strong Cox-2 expression and estrogen and progesterone receptor content of tumors ( P=0.037 and P=0.010). However, we could not demonstrate a significant association between Cox-2 staining and overall survival or disease free survival time. CONCLUSIONS: These results suggest that Cox-2 expression is significantly associated with less differentiated and more aggressive breast carcinomas and might therefore be a useful prognostic indicator as well as a target for therapy. PMID- 12884025 TI - p21WAF1/CIP1 is more effective than p53 in growth suppression of mouse renal carcinoma cell line Renca in vitro and in vivo. AB - PURPOSE: Although there are many controversial reports about the effect of p53 and p21(WAF1/CIP1) overexpression in different human tumor cells, the p53 gene is shown to be a more effective candidate for cancer gene therapy because of its more pronounced ability to induce apoptosis. In the present study, we present the effect of p53 and p21(WAF1/CIP1) overexpression on mouse renal carcinoma cells in vitro and in vivo. METHODS: p53 and p21(WAF1/CIP1) genes were introduced into Renca cells using adenoviral vectors (Ad5CMV-p53 and Ad5CMV-p21). The induction of apoptosis was measured using Annexin V assay and DNA fragmentation analysis. The expression of proteins was examined using immunocytochemistry and Western blot methods. The ability of adenoviral vectors to inhibit tumorigenicity of Renca cells, as well as the growth of pre-established tumors was measured. RESULTS: In vitro growth assays revealed higher growth suppression after Ad5CMV p21 infection. Although both vectors induced apoptosis, Ad5CMV-p53 was slightly more efficient. In vivo studies in Balb/c mice, demonstrated that tumorigenicity was completely suppressed by Ad5CMV-p21. Besides this, Ad5CMV-p21 significantly inhibited the growth of established tumors, while Ad5CMV-p53 did not. CONCLUSIONS: These data suggest that p21(WAF1/CIP1) is a more potent growth suppressor than p53 of mouse tumor cells Renca. The divergent responses of tumor cells to p21(WAF1/CIP1) overexpression could be due to various networks that differ between species. PMID- 12884026 TI - A small inhibitor of the interaction between Bax and Bcl-X(L) can synergize with methylprednisolone to induce apoptosis in Bcl-X(L)-overexpressing breast-cancer cells. AB - PURPOSE: To identify inhibitors of the interaction between Bax and Bcl-X(L). METHODS: Using an assay based on biosensor technology, we screened a chemical library of 10,000 compounds for inhibitors of the interaction between Bax and Bcl X(L). Using cell-culture systems we tested active compounds for their ability to induce apoptosis in Bcl-X(L)-overexpressing MCF7 cells and increase the sensitivities of the cells to apoptosis-inducing drugs [vincristine sulphate, dexamethasone, cycloheximide and 6alpha-methylprednisolone (MP)]. RESULTS: A single compound, 2',4',5',7'-tetrabromofluorescein (A5), from the library was found to inhibit this interaction efficiently. Several structural analogues of A5 were tested and two of these [4',5'-dibromofluorescein (A9) and 3,4,5,6 tetrabromofluorescein (A11)] were found to be active, and their activities were confirmed by an independent in vitro pull-down assay. These active compounds were observed to induce apoptosis in Bcl-X(L)-overexpressing MCF7 cells. Moreover, two of the compounds (A5 and A11) appeared to increase the sensitivities of the cells to MP. A more rigorous test using the isobologram technique showed that there is a synergistic cytotoxic effect between A11 and MP. CONCLUSIONS: We have identified a small inhibitor of the interaction between Bax and Bcl-X(L) that can synergize with methylprednisolone to induce apoptosis in Bcl-X(L)-overexpressing breast-cancer cells. PMID- 12884027 TI - Intra-mammary tumor location does not influence prognosis but influences the prevalence of axillary lymph-node metastases. AB - BACKGROUND: The number of axillary lymph-node metastases is not only a function of disease progression in primary breast cancer, but is also influenced by the intra-mammary location of the tumor. Nevertheless, the prognostic role of the tumor site is discussed controversially. The objective of this study was to analyze the impact of primary-tumor location on axillary lymph-node involvement, relapse, and mortality risk by univariate and multivariate analysis, in patients both with and without systemic and loco-regional treatment. METHOD: Retrospective analysis was conducted on 2,414 patients at the I. Frauenklinik, Ludwig Maximilians University, Munich and Berlin-Charlottenburg, who underwent R(0) resection of the primary tumor and systematic axillary lymph-node dissection (at least five lymph nodes resected) for UICC I-III-stage breast cancer. Patients with unknown tumor site, multifocal tumor spread, central tumor location, or tumor location within 15 degrees of the border between outer and inner quadrants were excluded from the study. Median observation time was 6.7 years. RESULTS: The primary tumor site was within or between the medial quadrants of the breast in 33.6% of the patients ( n=810) and in the lateral hemisphere of the breast in 66.4% ( n=1,604). Tumor size, histopathological grading, and estrogen receptor status were balanced between patients with lateral and medial tumor location. Metastatic axillary lymph-node involvement was significantly associated with a lateral tumor location ( P<0.0001). The mean number of axillary lymph-node metastases was increased by 29% in cases with lateral tumor location (2.2 vs 1.7, P=0.003). In a multivariate logistic regression analysis allowing for tumor location, estrogen receptor status, grading and tumor size, tumor location was confirmed as a significant risk factor ( P=0.02) for axillary lymph-node involvement. Tumor location, however, did not correlate with either disease-free survival (DFS) or overall survival (OS), by univariate (DFS: P=0.41; OS: P=0.57) or by multivariate analysis (DFS: P=0.16; OS: P=0.98). CONCLUSION: We conclude that there is no sufficient evidence to support any independent prognostic significance of intra-mammary tumor location in early breast cancer. However, medial tumor location may lead to the underestimation of axillary lymph-node involvement. PMID- 12884028 TI - Multivariate analysis of clinical prognostic factors in patients with glioblastoma multiforme treated with a combined modality approach. AB - We investigated the influence of various clinical prognostic factors in patients with glioblastoma multiforme (GBM) treated with a combined modality approach. A total of 175 patients with GBM was treated in four consecutive prospective phase II studies using surgery, hyperfractionated or accelerated hyperfractionated radiotherapy (RT) and either adjuvant or concurrent or pre-irradiation chemotherapy (CHT) between January 1988 and December 1993. The median survival time for all 175 patients was 14 months and 1-3-year survival (OS) rates were 57%, 34% and 24%, respectively. The median time to tumour progression was 12 months, and 1-3-year progression-free survival (PFS) rates were 43%, 11% and 7%, respectively. Survival analysis showed that of all investigated prognostic factors, only gender did not influence survival. Patients 55 years; patients with KPS 80-100 did better than those with KPS 50 70; patients with frontal tumours did better than those with tumours in other locations; patients with tumours up to 4 cm did better than those with larger tumours, as did patients with either subtotal or gross total tumour resection when compared to those undergoing biopsy only. Multivariate analysis showed that gender and tumour location did not independently influence survival. When PFS was used as the endpoint, only gender did not influence PFS, as confirmed by multivariate analysis. PMID- 12884029 TI - p34cdc2 expression is an independent indicator for lymph node metastasis in colorectal carcinoma. AB - PURPOSE: The significance of p34(cdc2) expression in human tumors has not been fully explained. The aim of the current study was to elucidate the clinicopathologic significance of immunohistochemical p34(cdc2) expression in carcinoma of the colon and rectum. METHODS: The immunohistochemical expression of p34(cdc2) was examined in 90 consecutive colorectal tumor cases, and p34(cdc2) expression and the clinicopathologic features of the patients and their tumors were compared. RESULTS: Lymph node metastasis was significantly more frequent in tumors expressing p34(cdc2) (47.8%, 11 of 23 tumors) than in tumors not expressing p34(cdc2) (22.4%, 15 of 67 tumors; P=0.020). Multivariate analysis demonstrated that tumor depth ( P=0.008) and p34(cdc2) expression ( P=0.022) were independently associated with lymph node metastases of colorectal carcinomas. CONCLUSIONS: The immunohistochemical expression of p34(cdc2) is independently associated with lymph node metastasis in colorectal carcinoma. PMID- 12884030 TI - Different apoptotic activity and p21(WAF1/CIP1), but not p27(Kip1), expression in serrated adenomas as compared with traditional adenomas and hyperplastic polyps of the colorectum. AB - PURPOSE: Serrated adenomas (SAs), which include a wide spectrum of lesions, can be broadly divided into two subtypes: type I, closely mimicking hyperplastic polyps (HPs), and type II, unequivocal adenomatous tumor. Our preliminary findings showed clinicopathologic differences between them. The present study was conducted to investigate apoptotic activity and expression of the cell cycle regulator proteins p21(WAF1/CIP1) and p27(Kip1) in type I and II SAs, as compared with traditional adenomas (TAs) and HPs. METHODS: Apoptotic activity was estimated in hematoxylin-eosin stained specimens, and p21(WAF1/CIP1) or p27(Kip1) immunoreactivity was determined in 62 SAs (19 type I and 43 type II), 50 TAs and 19 HPs. The numbers (percentages) of apoptotic or immunoreactive cells were counted per 1,000 epithelial cells in equally separated crypt zones (upper, middle, and lower thirds). RESULTS: The apoptotic activity in the middle, but not the upper or lower crypt zone was higher in type II SAs (median 0.2%, interquartile range 0.1-0.5%) than in HPs (0.1%, 0.1-0.2%, P<0.01), whereas it was lower in type I SAs (0.2%, 0.1-0.3%) than in TAs (0.5%, 0.2-0.6%, P<0.001). P21(WAF1/CIP1) expression in the lower crypt zone was higher in both type I and type II SAs (19.8%, 7.0-33.2% and 20.4%, 3.9-47.8%, P<0.0001) than in TAs (1.2%, 0.6-5.2%), and a similar tendency was also observed for the middle crypt zone. p27(Kip1) expression did not vary among the groups. CONCLUSIONS: The differences in apoptotic activity and p21(WAF1/CIP1) expression between SAs and TAs or HPs indicate that SA should be considered as a distinct subtype of colorectal neoplasm. The two subtypes of SA do not differ in these parameters despite specific clinicopathological features. PMID- 12884032 TI - Informed consent/assent in children. Statement of the Ethics Working Group of the Confederation of European Specialists in Paediatrics (CESP). AB - Informed consent means approval of the legal representative of the child and/or of the competent child for medical interventions following appropriate information. National legal regulations differ in regard to the question when a child has the full right to give his or her autonomous consent. Informed assent means a child's agreement to medical procedures in circumstances where he or she is not legally authorised or lacks sufficient understanding for giving consent competently. Doctors should carefully listen to the opinion and wishes of children who are not able to give full consent and should strive to obtain their assent. Doctors have the responsibility to determine the ability and competence of the child for giving his or her consent or assent. All children, even those not judged as competent, have a right to receive information given in a way that they can understand and give their assent or dissent. This consent/assent process must promote and protect the dignity, privacy and confidentiality of the child and his or her family. Consent or assent is required for all aspects of medical care, for preventive, diagnostic or therapeutic measures and research. Children may effectively refuse treatment or procedures which are not necessary to save their lives or prevent serious harm. Where treatment is necessary to save a life or prevent serious harm, the doctor has the duty to act in the best interest of the child. However, parents may also refuse to consent and in this case national laws and legal mechanisms for resolving disputes may be used. PMID- 12884031 TI - Double high-dose chemotherapy with adriamycin, paclitaxel, cyclophosphamide, and thiotepa followed by autologous peripheral blood stem cell transplantation in women with metastatic breast cancer. AB - To determine the feasibility, time to progression, and event-free survival, twenty-two women with metastatic breast cancer received two cycles of high-dose chemotherapy (HDCT) followed by peripheral blood stem cell transplantation (PBSCT) early after first-line induction chemotherapy. The median age of the ten (45.5%) pre- and 12 (54.5%) postmenopausal women was 48 (range: 33-60) years. Sixteen patients (72.7%) had at least two or more metastatic sites involved. Protocol induction and mobilization chemotherapy including granulocyte-colony stimulating-factor (G-CSF) consisted of two cycles with adriamycin (60 mg/m(2)) i.v. and paclitaxel (200 mg/m(2)) i.v. After collection of at least 4 x 10(6)/kg bodyweight peripheral blood stem cells, the first HDCT-course of adriamycin (60 mg/m(2)), paclitaxel (200 mg/m(2)) cyclophosphamide (4 g/m(2)), and thiotepa (800 mg/m(2)) (ATCT) was given to at least stable disease (SD) patients. Six to eight weeks later, the second HDCT-ATCT was administered. Each HDCT-cycle was followed by PBSCT with a median of 3.81 x 10(6)/kg bodyweight CD-34 positive cells (range: 1.85-10.38). All women showed median leukocyte engraftment (>1,000 x 10(9)/l) on day +9.4 (range: 7-13) and median platelet engraftment (>20,000 x 10(9)/l) on day +12.3 (range: 8-15). There were no apparent differences in the clinical course and non-hematologic toxicity between the two HDCT-cycles. Of the 21 patients evaluable for response, eight (38.1%) patients achieved complete remission (CR), ten (47.6%) patients showed a partial remission (PR), two patients (9.5%) no change, and one patient (4.8%) progressive disease. After a median observation time of 36 (range 28-55) months, six (28.6%) women are alive, four (19.0%) of them in continuous CR, including two women with stable bone lesions, respectively, and 15 (71.4%) died due to progressive disease. Median time to progression (TTP) was 8 (range 4-19) months. A high initial response rate of early HDCT, including the most active drugs adriamycin and paclitaxel, can be achieved with tolerable toxicity in metastatic breast cancer. New approaches for maintaining primary tumor response achieved with efficacious high-dose chemotherapy are warranted. PMID- 12884034 TI - An 8-year-old boy with a 4-day history of fever, cough and malaise, and a 2-day history of painful calves and difficulty walking. PMID- 12884033 TI - Encephalocraniocutaneous lipomatosis: a case with unilateral odontomas and review of the literature. AB - We report a 7-year-old boy with the typical features of encephalocraniocutaneous lipomatosis (ECCL) including unilateral skin, eye, bone, and asymptomatic central nervous system involvement. The presenting sign were numerous ipsilateral odontomas which have been described in only two previous patients so far. We reviewed all 32 cases reported to date and focus on the dermatological hallmark of the condition, namely a fatty tissue naevus of the scalp for which the term naevus psiloliparus was recently coined. Sporadic occurrence, patchy type of organ involvement and roughly balanced sex ratio in ECCL are compatible with the concept of a lethal autosomal mutation only surviving in a mosaic state. CONCLUSION: In children presenting with a congenital smooth hairless lesion on the scalp, the diagnosis of naevus psiloliparus should be considered and other signs of encephalocraniocutaneous lipomatosis should be looked for. PMID- 12884035 TI - Post-natal corticosteroid use. AB - Corticosteroids given systemically in the first 2 weeks after birth may reduce chronic lung disease (CLD) and mortality, but there are concerns regarding the long-term adverse effects which may have influenced prescribing habits. The aim of this study was to determine whether corticosteroids were still prescribed and, if so, was there a consensus regarding the most appropriate regimen to use. Consultant paediatricians at all 223 neonatal units in the United Kingdom were sent a questionnaire asking which route, timing, dosage and indications they used when administering corticosteroids. There was a 72% response rate to the survey. Of those who responded, 33% never prescribed steroids and 3.75% gave inhaled steroids only. Corticosteroids were not prescribed in the 1st week after birth, 21.1% of respondents commenced administration in the 2nd week and 33.3% only outside the neonatal period. A total of 48% prescribed a daily dose of 0.5 mg/kg per day, 33% gave lower doses, with 7.9% prescribing 0.1 mg/kg per day or less. The majority (77%) prescribed corticosteroids only to ventilator-dependent infants, others also gave corticosteroids to chronically continuous positive airway pressure- or oxygen-dependent infants. CONCLUSION: There are wide variations in corticosteroid usage. This emphasises the need to identify if there is a corticosteroid dosage regimen with a positive risk/benefit ratio. PMID- 12884036 TI - Experimental infection of dogs with Borrelia burgdorferi sensu stricto using Ixodes scapularis ticks artificially infected by capillary feeding. AB - Specific pathogen-free dogs were experimentally infected with Borrelia burgdorferi sensu stricto using nymphal or adult female Ixodes scapularis ticks artificially infected with spirochetes by capillary feeding. The ticks were capillary fed B. burgdorferi isolate 610, previously isolated from a dog with Lyme disease and grown in BSK medium. This isolate induced clinical signs in the dogs similar to those for dogs infested with ticks naturally infected with B. burgdorferi. Adult ticks were more efficient than nymphs in transmitting spirochetes to the dogs. One of five dogs infested with nymphal ticks capillary fed B. burgdorferi was skin biopsy culture and serologically positive, and demonstrated lameness. In contrast, all five dogs infested with adult female ticks that had been capillary fed with B. burgdorferi were culture and serologically positive, with one dog developing lameness. The immunoblot profiles of dogs challenged with female ticks infected by capillary feeding (8 weeks post challenge) were similar to immunoblots (4 weeks post challenge) from dogs challenged with naturally infected females collected in the field. These studies demonstrated that B. burgdorferi cultured in BSK medium can be capillary fed to either nymphal or adult female ticks under laboratory controlled conditions for the purpose of transmitting the spirochete to dogs during the tick's blood meal. This tick infection system would be useful for a controlled and defined challenge of vaccinated and non-vaccinated dogs for proper evaluation of vaccine efficacy, which is difficult to achieve using field-collected ticks. Furthermore, this system may also be useful for investigation of the pathogenesis of Lyme disease, evaluation of the pathogenicity of new isolates of B. burgdorferi, or evaluation of antibiotic therapy. PMID- 12884037 TI - Detection and quantification of Mycobacterium leprae in tissue samples by real time PCR. AB - Real-time PCR technology has improved molecular diagnostics of many pathogens, but no such test is available for Mycobacterium leprae. In this report we describe the establishment and the pre-clinical evaluation of such an assay. The test achieved a theoretical analytical sensitivity limit of 194 M. leprae cells per skin biopsy specimen and facilitated quantification of mycobacteria in tissue over a range of 54-54,000,000 cells per sample. In punch skin biopsies from 39 untreated Ugandan patients with newly diagnosed leprosy, the clinical diagnosis could be confirmed in 88.9% of multibacillary and 33.3% of paucibacillary (microscopically negative) patients. Real-time detection thus did not increase the clinical sensitivity of PCR as compared to conventional protocols, in spite of its evidently high analytical sensitivity. On the other hand, as still no culture system exists for M. leprae, the assay appears to be a robust tool for detection of the bacterium in selected clinical situations, as well as for quantitation in experimental settings. PMID- 12884038 TI - The role of graft-resident Kupffer cells and lymphocytes of donor type during the time course after liver transplantation--a clinico-pathological study. AB - Although graft-resident passenger leukocytes are known to mediate acute rejection by triggering direct allorecognition, they may also act in an immunomodulatory fashion and play an important role in tolerance induction. The present study evaluated by non-isotopic in situ hybridization and immunohistochemistry if and during which time period after transplantation Kupffer cells (KC) and lymphocytes of the liver were replaced by recipient cells and if there is any correlation with the occurrence of rejection episodes and the clinical course. A successive re-population of the liver by recipient lymphocytes and KC was observed after transplantation but a smaller portion of lymphocytes and KC with donor genotype was detectable during the whole time course studied. There was no correlation between the portion of recipient-derived KC and donor-derived lymphocytes and histopathological alterations of the liver tissue. The biopsy content of KC with recipient origin has had no prognostic significance for the probability of survival, but patients with a low portion of donor lymphocytes in the liver biopsy obtained during the first week after transplantation have had a better prognosis for survival. The present results indicate that graft-resident KC and lymphocytes are potentially not the main cell types involved in tolerance induction after liver transplantation. PMID- 12884039 TI - Three-dimensional observations on the alterations of lobular architecture in chronic hepatitis with special reference to its angioarchitecture for a better understanding of the formal pathogenesis of liver cirrhosis. AB - For a better understanding of the formal pathogenesis of liver cirrhosis, the angioarchitecture of the liver lobule in chronic viral hepatitis was investigated three dimensionally. The histological reconstruction method, using serial histological sections, was adopted for the three-dimensional observation. Histology of the case showed chronic active hepatitis with occasional fibrous bridging of the portal to portal tract or hepatic vein. Graphic reconstructions revealed various degrees of altered angioarchitecture from place to place. While the conducting portion of the portal vein was almost preserved, the pathological changes mostly began at the parenchymal portion, especially second step or subsequent branches of the portal vein. In general, portal vein branches showed damage such as stenoses, disappearance, an increase and decrease in number and distorted spatial arrangements. Even in less damaged portal tracts, portal veins showed such changes to some extent. In severely damaged places with bridging fibrosis, a normal lobular angioarchitecture was completely lost; instead, portal veins, arteries and hepatic veins were tangled with each other. Parenchymal nutrition was suggested to be dependent on the remaining third-step portal branches or newly formed ones. However, the hepatic vein system had a tendency to be preserved and distributed fluently in the parenchyma. The distortion of these portal vessels indicated various degrees of loss of the lobular architecture. In conclusion, it is suggested that an early histological sign of cirrhosis develops in the course of chronic hepatitis. PMID- 12884040 TI - Important role for fibronectin-EIIIA during renal tubular repair and cellular recovery in uranyl acetate-induced acute renal failure of rats. AB - The present study was designed to identify the source and kinetics of an alternatively spliced "embryonic" cellular fibronectin EIIIA (cFn-EIIIA) in relation to regenerating renal tubules in uranyl acetate (UA)-induced acute renal failure (ARF) in rats. Damage of the proximal tubules was found as early as day 2 after induction of ARF, peaked at day 5, and was almost substituted by epithelial relining by day 7. Immunohistochemistry showed de novo deposition of cFn-EIIIA in peritubular regions as early as day 2, then on the tubular basement membrane (TBM) after day 4. beta1 Integrin, the receptor for Fn, was mainly found at the basal side of tubules in the normal control and increased in the interstitium after induction of ARF, but the staining pattern gradually returned to the control after day 7. Immunoelectron microscopy revealed that cFn-EIIIA was produced initially by the peritubular endothelium and later by fibroblastic cells and was deposited to the TBM, on which regenerating tubules proliferated, probably with cFn-EIIIA production. beta1 Integrin was expressed in cFn-EIIIA producing cells, especially in regenerating tubular cells, suggesting that cFn EIIIA signal transduction affects regenerating tubules. Transforming growth factor (TGF)-beta1 was found in some damaged proximal tubules and interstitial cells after induction of ARF and later in the regenerating tubules. CFn-EIIIA and beta1 integrin mRNA levels were upregulated as early as day 2. TGF-beta1 mRNA level significantly increased after day 3, suggesting a modulatory role for TGF beta1 on cFn-EIIIA production, but not by day 2. Our data suggest that cFn-EIIIA production by the endothelium during the very early response to tubular injury and by fibroblastic cells and regenerating tubules may play an important role in the cellular recovery of UA-induced ARF in rats. PMID- 12884042 TI - Regulation of proton-to-electron stoichiometry in photosynthetic electron transport: physiological function in photoprotection. AB - The primary stable products of photosynthetic electron flow are NADPH and ATP. Stoichiometry of their production depends on the ratio of protons pumped across the thylakoid membrane to electrons passed through the electron transport pathway (H(+)/e(-) ratio). Flexible requirements of the ATP/NADPH ratio by various assimilatory reactions in chloroplasts must be fulfilled by the H(+)/e(-) ratio during the electron flow. In addition to the well-known role of Delta pH during ATP synthesis, Delta pH also functions as a trigger of the down-regulation of photosystem II (PSII) photochemistry. Excessive light energy is safely dissipated as heat by this regulatory process to suppress the generation of toxic reactive oxygen species. Thus, regulation of the H(+)/e(-) ratio may function in the photoprotection, as well as in the regulation of the ATP/NADPH production ratio. It has long been the consensus that the H(+)/e(-) ratio can be controlled by regulating the proton-transporting Q-cycle in the cytochrome b(6)f complex and by the cyclic electron flow around photosystem I (PSI). Despite the possible physiological importance and the long history of interest, the molecular identity of Q-cycle regulation and the cyclic electron flow around PSI have been remained unclear. The recent improvements in research tools, including the genetic approach using chlorophyll fluorescence imaging and establishment of the chloroplast transformation technique, are providing new insights into classical topics. In this review, we focus on regulation of the H(+)/e(-) ratio especially from the view of photosynthetic regulation. PMID- 12884043 TI - Genetic diversity and origin of Potamogeton anguillanus (Potamogetonaceae) in Lake Biwa, Japan. AB - We analyzed the genetic variation in Potamogeton anguillanus Koidz. and its putative parents, P. malaianus Miq. and P. perfoliatus L., at five allozyme loci of four enzymes to test the hypothesis of a hybrid origin for P. anguillanus, collected in Lake Biwa, Japan. Alleles diagnostic for either P. malaianus or P. perfoliatus were present at four loci. Of 13 single locus phenotypes (SLPs) of P. anguillanus, eight were phenotypes that were expected in F(1) hybrids between P. malaianus and P. perfoliatus. Two SLPs were different from those expected in F(1) hybrids but could have resulted from segregation of parental alleles in later generation hybrids. Each of the remaining three SLPs possessed one allele unique to P. anguillanus. Allozyme analyses thus supported the view that P. anguillanus was derived from hybridization between P. malaianus and P. perfoliatus. It seems likely that the genetic diversity of P. anguillanus found previously originated through multiple hybridizations and sexual processes in P. anguillanus. Other processes such as intragenic recombination, mutation, or hybridization with another lineage are also discussed with reference to the origin of unique alleles. PMID- 12884041 TI - Distribution of p63, cytokeratins 5/6 and cytokeratin 14 in 51 normal and 400 neoplastic human tissue samples using TARP-4 multi-tumor tissue microarray. AB - p63, cytokeratin (CK) 5/6 and CK 14 have been employed in diagnostic pathology as markers of basal, squamous and myoepithelial differentiation in several types of human neoplasms; however, there is scant data on the concurrent expression of these markers in large series of human neoplasms. We analyzed the distribution of these three immunohistochemical markers in 51 normal human tissue samples, 350 carcinomas, 25 malignant melanomas (MMs), and 25 glioblastomas using three serial sections of tissue array research program (TARP)-4 multi-tumor tissue microarray. Also, we performed double immunostainings to characterize the differential distribution of p63/CK 5/6 and p63/CK 14 in normal breast, salivary gland and skin. p63, CK 5/6 and CK 14 were expressed in basal cells of the prostate and respiratory epithelia and in breast and bronchial myoepithelial cells. p63 was also expressed in cytotrophoblast cells of human placenta and in scattered cells of lymph node germinal center. CK 5/6 and CK 14 also stained the cytoplasm of basal cells of esophageal stratified squamous epithelium and transitional epithelial cells of the bladder. No mesenchymal, neural, endothelial, smooth muscle or adipose cells were stained by any of the markers. p63, CK 5/6, and CK 14 were respectively expressed in 92.6%, 75.0%, and 52.9% of the squamous cell carcinomas of the lung, 10.2%, 20.0%, and 7.4% of the ductal carcinomas of the breast, 12.9%, 34.4%, and 11.8% of the serous and 25.0%, 0%, and 0% of the endometrioid carcinomas of the ovary. Lung, prostate and colonic adenocarcinomas, as well as MMs and glioblastomas were only rarely decorated by one of the markers. Only matched samples of 16 squamous cell carcinomas and two ductal carcinomas of the breast co-expressed these three markers. In double immunostainings, p63-CK 5/6, as well as p63-CK 14 were co-expressed by basal/myoepithelial cells of the salivary glands and basal cells of the epidermis. Our results demonstrate that p63, CK 5/6 and CK 14 may be used together in immunohistochemical panels to characterize squamous differentiation in poorly differentiated carcinomas or carcinomas of unknown origin. PMID- 12884044 TI - Cytological study of Tibetia (Fabaceae) in the Hengduan Mountains region, China. AB - The Hengduan Mountains comprise one of the world's most important hot spots of biodiversity. Tibetia (Ali) H.P. Tsui (Fabaceae), which has four or five species in two sections, is one of the genera endemic to the region. This paper describes for the first time the karyotype of three of those species. The chromosome counts of all three are 2 n = 16. The karyotypes of the species examined contain chromosomes of variable karyotypic symmetry with centromeres at median and submedian positions that correlate with the morphological characteristics of the species. Karyotypic variation at the diploid level appears to be the predominant feature of chromosome evolution in the genus and may provide a clue to the study of evolutionary patterns of plants in this region. PMID- 12884045 TI - Growth promotion and an increase in cell wall extensibility by silicon in rice and some other Poaceae seedlings. AB - The effect of silicon on organ growth and its mechanisms of action were studied in rice ( Oryza sativa L. cv. Koshihikari), oat ( Avena sativa L. cv. Victory), and wheat ( Triticum aestivum L. cv. Daichino-Minori) seedlings grown in the dark. Applying silicon in the form of silicic acid to these seedlings via culture solution resulted in growth promotion of third (rice) or second (oat and wheat) leaves. The optimal concentration of silicon was 5-10 mM. No growth promotion was observed in early organs, such as coleoptiles or first leaves. In silicon-treated rice third leaves, the epidermal cell length increased, especially in the basal regions, without any effect on the number of cells, showing that silicon promoted cell elongation but not cell division. Silicon also increased the cell wall extensibility significantly in the basal regions of rice third leaves. These results indicate that silicon stimulates growth of rice and some other Poaceae leaves by increasing cell wall extensibility. PMID- 12884046 TI - Genetic variation in populations of the morphologically and ecologically variable fern Stegnogramma pozoi subsp. mollissima (Thelypteridaceae) in Japan. AB - In Japanese Stegnogramma pozoi subsp. mollissima (Fisher ex Kunze) K. Iwats. there is the intrasubspecific variation among rbcL sequences. Northern and southern plants are genetically differentiated for maternally inherited cpDNA. In the present study we examined allozyme polymorphisms to test the hypothesis that northern and southern plants may be separate species. Based on allozyme data, the degree of gene flow among populations was estimated to be large. The artificial crossing experiments between cpDNA haplotypes also suggested that isolation has not developed among these cpDNA haplotypes. However, interpopulation genetic differentiation in cpDNA was observed even in the small area at the foot of Mt. Hakone, and the cpDNA haplotypes appear to have different habitat preferences. PMID- 12884047 TI - Tension wood fibers are related to gravitropic movement of red mangrove (Rhizophora mangle) seedlings. AB - Freshly collected viviparous seedlings (propagules) were collected from wild plants of Rhizophora mangle and planted in vertical or horizontal positions. A total of 80 seedlings were examined anatomically at various ages and orientations. After rooting, seedlings reoriented from horizontal to vertical by extreme bending in the hook region of the hypocotyl directly above the basal 1 cm where roots formed. Hypocotyl bending occurred over many months. Trends in position and relative abundance of tension fibers (also called gelatinous fibers) over time were followed. The erection of the seedling was related to increased secondary xylem and the number of tension wood fibers on the upper side of the hook region. However, linear regressions had low coefficient of determination (r(2)) values, presumably related to seedlings with high variability. PMID- 12884048 TI - Floral scent chemistry of mangrove plants. AB - The flowers of mangrove plants are pollinated by a variety of pollinators including birds, bats, and insects. This study analyzed the floral scent chemistry of mangroves on Iriomote Island (located near Taiwan) including Bruguiera gymnorrhiza (L.) Lamk. (Rhizophoraceae), Kandelia candel (L.) Druce (Rhizophoraceae), Rhizophora stylosa Griff. (Rhizophoraceae), Sonneratia alba J. Smith (Sonneratiaceae), Nypa fruticans (Thunb.) Wurmb. (Palmae), Lumnitzera racemosa Willd. (Combretaceae), Avicennia marina (Forsk.) Vierh. (Avicenniaceae or Verbenaceae), and Pemphis acidula Forst. (Lythraceae). A total of 61 chemicals (fatty acid derivatives, terpenoids, carotenoid derivatives, benzenoids, nitrogen containing compounds, 13 unknown chemicals) were detected in the floral scents of the various species. The species displayed a distinct chemical profile ranging from only two chemicals in the floral scent of Kandelia candel to more than 25 chemicals in the floral scent of Nypa fruticans. All of the identified chemicals have been found in the floral scents of other angiosperms. The chemical profile of some species can be correlated with their floral morphology and pollinators. PMID- 12884049 TI - Temperature dependence of seedling establishment of a perennial, Dioscorea tokoro. AB - The temperature dependence of seed germination and seedling growth was analyzed in Dioscorea tokoro, an East Asian summer-green perennial. Seeds were able to germinate fully only at 11-20 degrees C. At around 17-20 degrees C the first leaf petiole of the seedling elongated and quickly set the first leaf blade at a position enabling photosynthesis. At temperatures higher than 20 degrees C petiole elongation was retarded, and seedlings formed a rhizome and established as a perennial. The rhizome size increased with temperature up to 29 degrees C. Thus, during growth immediately after germination, temperature appears to be a key factor in determining whether the plant establishes as a perennial or grows rapidly without rhizome thickening. PMID- 12884050 TI - Optical and anatomical characteristics of bracts from the Chinese "glasshouse" plant, Rheum alexandrae Batalin (Polygonaceae), in Yunnan, China. AB - Bracts that lacked chlorophyll were compared with rosette leaves on the Chinese glasshouse plant Rheum alexandrae Batalin. The structures were analyzed anatomically and with photospectrometry. Histological features were significantly different between the bracts and the rosette leaves. Epidermal pavement cells and palisade cells were larger in the bracts than in the rosette leaves, but the ratio of the intercellular spaces to the cells in the palisade layer was the same in the two structures. Absorption spectrum analyses of the half-translucent bracts showed that, despite their thinner structure, they absorbed ultraviolet (UV) radiation more efficiently than did the green rosette leaves. The characteristics of the R. alexandrae bracts were different from those of the Himalayan glasshouse plant Rheum nobile. These results suggest that R. alexandrae evolved bracts as an adaptive strategy to protect the inflorescence from UV radiation. PMID- 12884051 TI - Development of primer sets for PCR amplification of the PgiC gene in ferns. AB - Polymerase chain reaction (PCR)-based nuclear DNA markers were developed for fern species. We first determined the partial nucleotide sequence of cDNA of the pgiC gene encoding cytosolic phosphoglucose isomerase from Dryopteris caudipinna, and then PCR primers for exon-primed, intron-crossing (EPIC) amplifications were designed. The EPIC primers are universally applicable to the most derived indusiate fern families such as Dryopteridaceae, Thelypteridaceae, and Woodsiaceae. The PCR products of primers 14F/16R containing two introns are moderate in size (534 bp-ca.1000 bp) and are possibly of value in phylogenetic reconstruction at specific and generic levels. Codominant nuclear DNA markers applicable to the estimation of mating systems and other population genetic studies were also developed by a combination of single-strand conformation polymorphism (SSCP) and EPIC amplification using primers 14F/15R and 15F/16R. In order to provide a case study using these markers, allelic variation of PCR products using 15F/16R was examined in populations of Arachniodes standishii (Dryopteridaceae). PMID- 12884052 TI - Alpha1-adrenergic signaling mechanisms in contraction of resistance arteries. AB - Our goal in this review is to provide a comprehensive, integrated view of the numerous signaling pathways that are activated by alpha(1)-adrenoceptors and control actin-myosin interactions (i.e., crossbridge cycling and force generation) in mammalian arterial smooth muscle. These signaling pathways may be categorized broadly as leading either to thick (myosin) filament regulation or to thin (actin) filament regulation. Thick filament regulation encompasses both "Ca(2+) activation" and "Ca(2+)-sensitization" as it involves both activation of myosin light chain kinase (MLCK) by Ca(2+)-calmodulin and regulation of myosin light chain phosphatase (MLCP) activity. With respect to Ca(2+) activation, adrenergically induced Ca(2+) transients in individual smooth muscle cells of intact arteries are now being shown by high resolution imaging to be sarcoplasmic reticulum-dependent asynchronous propagating Ca(2+) waves. These waves differ from the spatially uniform increases in [Ca(2+)] previously assumed. Similarly, imaging during adrenergic activation has revealed the dynamic translocation, to membranes and other subcellular sites, of protein kinases (e.g., Ca(2+)-activated protein kinases, PKCs) that are involved in regulation of MLCP and thus in "Ca(2+) sensitization" of contraction. Thin filament regulation includes the possible disinhibition of actin-myosin interactions by phosphorylation of CaD, possibly by mitogen-activated protein (MAP) kinases that are also translocated during adrenergic activation. An hypothesis for the mechanisms of adrenergic activation of small arteries is advanced. This involves asynchronous Ca(2+) waves in individual SMC, synchronous Ca(2+) oscillations (at high levels of adrenergic activation), Ca(2+) sparks, "Ca(2+)-sensitization" by PKC and Rho-associated kinase (ROK), and thin filament mechanisms. PMID- 12884053 TI - Retrospective analysis of spinal missile injuries. AB - One hundred six patients with spinal missile injury from war zones were admitted to our department from 1994 to 2000. Functional recovery and complications in surgical and conservative treatment groups were evaluated. Sixty-five were treated surgically, of whom 55 (84%) had incomplete injuries (Frankel scores B, C, and D). In the conservative group, 28 (68%) had incomplete injuries. A total of 81 patients (53 in the surgical group, 28 in the conservative group) could be monitored for functional recovery. In the surgical group, 34 (64%) showed improvement, 15 (28%) were unchanged, and four (7%) worsened. In the conservative group, 17 (60%) improved, nine (32%) remained unchanged, and two (7%) worsened. Cerebrospinal fluid fistula was observed in ten patients, seven of them in the surgically treated group. Five of seven meningitides were seen in the surgically treated group. Surgical intervention is not essential for spinal gunshot injury; however, it may be beneficial for patients with CSF fistula, infectious and compressing foreign bodies in the injury site, instability, and rapid neurological deterioration. PMID- 12884054 TI - Effects of mexiletine, ginkgo biloba extract (EGb 761), and their combination on experimental head injury. AB - Lipid peroxidation (LP) and brain edema are important factors that produce tissue damage in head injury. The purpose of this study was to investigate the effect of mexiletine, gingko biloba extract (EGb 761), and their combination on LP and edema after moderate head trauma. Forty rats were randomly and blindly divided into four groups of ten animals each: control group (bolus injection of physiological saline), mexiletine group (50 mg/kg per injection), EGb 761 group (30 mg/kg per injection), and mexiletine plus EGb 761 group (50 mg/kg and 30 mg/kg per injection, respectively). The injections were given intraperitoneally at 1 h, 9 h, and 17 h after trauma. Twenty-four hours after injury, the rats were killed, and malondialdehyde (MDA) levels and brain water content were determined. Rats treated with mexiletine, EGb 761, and mexiletine plus EGb 761 had significantly lower MDA levels than the control group (P<0.01). The lowest MDA levels were measured in the mexiletine plus EGb 761 group. However, there was no significant difference in brain water content between treated groups and the control group (P>0.05). These findings show the usefulness of mexiletine and its combination with EGb 761 as a cerebroprotective agent in this model of experimental head injury. PMID- 12884055 TI - Hyperostotic paraganglioma of occipitotemporal bone. AB - A 25-year-old male presented with off-and-on vertigo of 10-year duration. He had left-sided cerebellar signs, left vocal cord paresis, and minimal left-sided hearing impairment. Computed tomography of the head revealed significant hyperostosis of squamous occipital, mastoid, and petrous temporal bone with no adjacent soft tissue mass. On excision, it turned out to be paraganglioma. Paraganglioma as a pure bony mass is not reported in the literature. The site of origin of such a tumor could not be ascertained, even on generous screening. The tumor remained nonsecretory on clinical and biochemical investigations. PMID- 12884056 TI - Single cerebral metastasis from colorectal adenocarcinoma. AB - Single cases are described in 50% of reported intracranial metastases. Single cerebral metastasis from colorectal adenocarcinoma is not very common, with a frequency varying between 0.5% and 1%. In our institute between 1960 and 2000, 44 patients affected by single metastasis from colorectal carcinoma were surgically treated. Surgical treatment with postoperative radiant therapy is necessary. These patients show improved quality of life, above all in relation to the maintenance of functional autonomy during the survival period. PMID- 12884057 TI - Statistical issues and methods for meta-analysis of microarray data: a case study in prostate cancer. AB - With the proliferation of related microarray studies by independent groups, a natural step in the analysis of these gene expression data is to combine the results across these studies. However, this raises a variety of issues in the analysis of such data. In this article, we discuss the statistical issues of combining data from multiple gene expression studies. This leads to more complications than those in standard meta-analyses, including different experimental platforms, duplicate spots and complex data structures. We illustrate these ideas using data from four prostate cancer profiling studies. In addition, we develop a simple approach for assessing differential expression using the LASSO method. A combination of the results and the pathway databases are then used to generate candidate biological pathways for cancer. PMID- 12884058 TI - Herman Jan Phaff: professor, mentor, friend and colleague. AB - Herman Jan Phaff, the father of yeast ecology, was born in the Netherlands in 1913. In his early years, he spent much time in his family's winery, which sparked his interest in microbes. Trained in the famous Delft tradition, Phaff discovered many unrecognized ecological niches of yeast, such as shellfish, rabbit stomach, frass of bark beetles, tree exudates, cactus roots, Capri figs, sewage, Drosophila flies and shrimp. He is also remembered for his pioneering work on the coevolution of yeasts, insects and plants as well as for his work on yeast beta-glucanase, which resulted in major advances in the understanding of the nature of the yeast cell wall. Phaff's legacy includes research on pectin degradation by fungal enzymes and the application of molecular approaches to yeast systematics. He discovered and described many yeasts, such as the yeast named in his honor, Phaffia rhodozyma, which led to the establishment of a very important industrial fermentation process yielding high concentrations of the pigment astaxanthin, used throughout the world to provide a natural source of this important carotenoid. PMID- 12884059 TI - The Phaff school of yeast ecology. AB - Herman Jan Phaff's legacy includes pioneering work on the yeast cell envelope and the application of molecular approaches to yeast systematics. Clearly, his interest and knowledge spanned the whole gamut of yeast biology. Yet, his most original and most heartfelt contribution was to our understanding of the position occupied by yeasts in nature. This view developed through the juxtaposition of his childhood exposure to industrial fermentations and his training in the tradition of Beijerinck's Delft School of Microbiology. Through some of Phaff's recent writings, I have attempted to formulate the themes or principles that were implicit to his ecological thinking. Six focal points emerge. (1) Yeasts in themselves are a sufficient object of study. (2) A clear idea of a yeast community cannot be obtained unless the yeast species are correctly identified. (3) Ecologically meaningful conclusions require an adequate sample size. (4) The bacteriological dictum "everything is everywhere" is a poor account of yeast distributions. (5) The habitat is the cornerstone of yeast ecology. (6) Ecology is the most exciting aspect of yeast biology. PMID- 12884060 TI - Evaluation of the BD Phoenix automated identification and susceptibility testing system in clinical microbiology laboratory practice. AB - The Phoenix Automated Microbiology System (BD Biosciences, USA) is a new, fully automated system for the rapid identification and antimicrobial susceptibility testing of gram-positive and gram-negative bacteria. The objective of this study was to evaluate the quality of performance of the Phoenix system in the identification and antimicrobial susceptibility testing of 260 gram-negative ( n=174) and gram-positive ( n=86) isolates collected from Polish hospitals in recent years. Two Phoenix panel types for identification/antimicrobial susceptibility testing, NMIC/ID-5 for gram-negative rods and PMIC/ID-4 for gram positive cocci, were used in the analysis according to the manufacturer's recommendations. The results produced by the system were compared with data obtained by reference or conventional microbiological methods. A high rate of agreement between the Phoenix and the conventional methods was observed for identification, ranging from 100% for gram-positive cocci to 96.0% for gram negative nonfermenters and 92.5% for members of the family Enterobacteriaceae. Similarly, a high level of agreement characterized the antimicrobial susceptibility data obtained with the Phoenix and by the agar dilution method (2,361 test results). For staphylococci, enterococci and Enterobacteriaceae, the methods were 100% concordant in determining the category of susceptibility of isolates to the majority of the antimicrobial agents tested. A category agreement value of below 90% was found for the susceptibility of enterococci and gram negative nonfermenters to ciprofloxacin (84.6% and 88.5%, respectively) and for susceptibility of Stenotrophomonas maltophilia to trimethoprim-sulfamethoxazole (80.0%). PMID- 12884061 TI - Community-acquired Klebsiella pneumoniae central nervous system infections in adults in Singapore. AB - A series of successfully managed community-acquired Klebsiella pneumoniae infections of the central nervous system (CNS) that occurred in six non neurosurgical adult patients in Singapore over a 2-year period is reported. Three patients had abscesses that were drained, and three had meningitis; all were treated with third-generation cephalosporins. All six patients were middle-aged adults, three of whom were food handlers. Clinicians should be aware of Klebsiella pneumoniae as an important CNS pathogen, particularly in East Asia. Although third-generation cephalosporins have had a major impact on the outcome of Klebsiella pneumoniae CNS infections, the emergence of extended-spectrum beta lactamase-producing strains may lead to their reduced efficacy in this clinical setting. PMID- 12884062 TI - Influence of mistletoe ( Viscum album) extracts on phagocytosis/burst activity of human phagocytes. AB - The aim of the present study was to examine the in vitro effects of various mistletoe extracts on human phagocytes. Phagocytosis, oxidative burst, and intracellular killing were analyzed by flow cytometry. None of the mistletoe preparations investigated exhibited significant phagocytosis-enhancing properties. Different mistletoe lectin concentrations ranging from 0.025 to 20 ng/ml showed only marginal influence on phagocyte activity. Contrary to the opinion of numerous practitioners of complementary medicine, the mistletoe preparations tested do not enhance phagocytic function. PMID- 12884063 TI - High incidence of gram-negative bacillary infection and high mortality in adult patients with bacterial meningitis and nasopharyngeal carcinoma. PMID- 12884064 TI - Characterisation of methicillin-resistant Staphylococcus aureus with reduced susceptibility to teicoplanin in Eastern France. PMID- 12884065 TI - Comparative evaluation of BDProbeTec ET, LCx and PACE 2 assays for the detection of Chlamydia trachomatis in urogenital specimens. PMID- 12884067 TI - Group a streptococcal serotypes isolated from healthy schoolchildren in iran. AB - Serotypes of group A streptococci are still a major cause of pharyngitis and some post-infectious sequelae such as rheumatic fever. As part of the worldwide effort to clarify the epidemiological pattern of group A streptococci in different countries, the present study was conducted to assess the prevalence of Streptococcus pyogenes serotypes in Iran. A total of 1588 throat swabs were taken from healthy school children in the city of Gorgan during February and March 1999. Of those isolates, 175 resulted positive for group A streptococci. The distribution pattern was similar for girls and boys, with 10.8% and 11.2%, respectively. Urban school children showed a higher rate of colonization compared to those in rural areas. Serotyping was performed on 65 of the positive isolates using standard techniques, and only 21 (32%) were M-type isolates. Their profiles fell into four types with M1 predominating, which could reflect the presence of rheumatic fever in the region. However, when isolates were challenged for T antigen types, nearly all were positive (94%). The pattern of T types was diverse (18 types), with the most common T types being T1 (26%), TB3264 (15%), TB?1-19 & B?25?1-19 (9.2%) and T2 & 2?28 (7.7%). When isolates were tested for opacity factor, only 23 (35%) were positive while 34 (52%) responded to the serum opacity reaction test. Although the number of isolates in this study was not sufficient to make any epidemiological conclusions, the scarcity of serotyping studies in Iran could render these data useful for future attempts to develop a streptococcal vaccine. PMID- 12884066 TI - Staphylococcus aureus colonization in a community sample of HIV-infected and HIV uninfected drug users. AB - HIV-infected individuals, especially those with a history of injecting drug use, are at high risk of Staphylococcus aureus infection. Moreover, the use of antimicrobial agents for opportunistic infections may increase nasal colonization by antimicrobial-resistant Staphylococcus aureus in this population and, subsequently, levels of infection with multidrug-resistant Staphylococcus aureus in the community. Between February 1999 and March 2000, 500 subjects from a community-based cohort of drug users completed an interview and underwent a physical exam. Risk factors for colonization by Staphylococcus aureus were examined, the antibiotic susceptibility profiles of all strains were determined, and DNA strain analysis was performed. One hundred twenty (24%) subjects had positive Staphylococcus aureus nasal cultures. Only HIV infection and homelessness were associated with Staphylococcus aureus colonization. Ten (8%) isolates were methicillin-resistant Staphylococcus aureus. Methicillin-resistant Staphylococcus aureus isolates were found more frequently among HIV-infected than HIV-uninfected respondents (14% vs. 3%, P=0.04). Among those colonized and HIV infected, the mean number of resistant isolates was higher for those currently reporting antibiotic use (5.0 vs. 2.3, P<0.001) and for those with CD4+ counts 1 IU/ml) was detected in 100%. However, only 3.3% of 20-year-old participants who did not know whether they had been vaccinated as children or young adults had a protective antibody level. After administering a single dose of tetanus vaccine to this group, the rate of protective antibody increased from 3.3% to 96.7%. For the prevention of tetanus, the optimal policy should focus on improving rates of complete primary immunization in adults and children and giving a single booster every 10 years, or at least at age 40 or 50 years. PMID- 12884074 TI - Genomovar diversity amongst Burkholderia cepacia complex isolates from an Australian adult cystic fibrosis unit. AB - In this study, a combination of recA-based PCR assays and 16S rDNA restriction fragment length polymorphism (RFLP) analysis was used to determine the genomovar diversity of clinical Burkholderia cepacia complex isolates. Twenty-eight isolates were prospectively collected from patients attending a large Australian adult cystic fibrosis (CF) unit, 22 isolates were referred from other Australian CF units and a further eight isolates originated from patients without CF. The 28 prospectively collected isolates were distributed amongst the following genomovars: Burkholderia cepacia genomovar I (28.6%), Burkholderia multivorans (21.4%), Burkholderia cepacia genomovar III (39.3%), Burkholderia vietnamiensis(3.6%) and Burkholderia ambifaria (7.1%). The results of this study highlight the usefulness of 16S rDNA RFLP typing for the identification of other Burkholderia spp. and non-fermenting gram-negative bacteria. PMID- 12884075 TI - Molecular and serological characterization of group a rotavirus isolates obtained from hospitalized children in Goiania, Brazil, 1998-2000. AB - Fecal samples positive for rotavirus group A ( n=120) were analyzed by enzyme immunoassay-monoclonal antibody (EIA-MAb) serotyping and/or reverse transcription/multiplex polymerase chain reaction (PCR) amplification to determine the prevalence of the [P] and G genotypes. The most prevalent G genotype/serotype detected was G1 (76.7%), followed by G2 (5.0%). Six samples were characterized as G9 by multiplex PCR, and one sample was characterized as G3 by EIA-MAb. The combinations of [P] and G genotypes found were P[8] and G1 (20.8%), P[6] and G1 (10.8%), P[6] and G9 (4.2%), P[8] and G2 (1.7%), and P[6] and G2 (0.8%). The diversity of rotavirus group A [P] and G genotypes/serotypes reinforces the need for continuous characterization of rotaviruses circulating in populations in Brazil. PMID- 12884076 TI - Severe disseminated tuberculosis after intravesical instillation of Bacillus Calmette-Guerin. PMID- 12884077 TI - Primates in cyberspace: using interactive computer tasks to study perception and action in nonhuman animals. PMID- 12884078 TI - Tufted capuchins (Cebus apella) attribute value to foods and tools during voluntary exchanges with humans. AB - This research examined exchange and value attribution in tufted capuchin monkeys ( Cebus apella). We presented subjects with opportunities to obtain various foods and a tool from an experimenter in exchange for the foods or tool in the subjects' possession. The times elapsed before the first chow biscuits were expelled and/or an exchange took place were recorded as the dependent measures. Laboratory chow biscuits, grapes, apples, and a metal bolt (a tool used to probe for syrup) were used as experimental stimuli. The subjects demonstrated the ability to recognize that exchanges could occur when an experimenter was present with a desirable food. Results indicate that subjects exhibited significant variation in their willingness to barter based upon the types of foods that were both in their possession and presented by the experimenter. Subjects more readily traded chow biscuits for fruit, and more readily traded apples for grapes than grapes for apples. During the exchange of tools and food, the subjects preferred the following in descending order when the probing apparatus was baited with sweet syrup: grapes, metal bolts, and chow biscuits. However when the apparatus was not baited, the values changed to the following in descending order: grapes, chow, and metal bolts. These results indicate that tufted capuchins recognize opportunities to exchange and engage in a simple barter system whereby low-valued foods are readily traded for more highly valued food. Furthermore, these capuchins demonstrate that their value for a tool changes depending upon its utility. PMID- 12884079 TI - Representation of two geometric features of the environment in the domestic chick ( Gallus gallus). AB - We report experiments based on a novel test in domestic chicks ( Gallus gallus), designed to examine the encoding of two different geometric features of an enclosed environment: relative lengths of the walls and amplitude of the corners. Chicks were trained to search for a food reward located in one corner of a parallelogram-shaped enclosure. Between trials, chicks were passively disoriented and the enclosure was rotated, making reorientation possible only on the basis of the internal spatial structure of the enclosure. In order to reorient, chicks could rely on two sources of information: the relative lengths of the walls of the enclosure (associated to their left-right sense order) and the angles subtended by walls at corners. Chicks learned the task choosing equally often the reinforced corner and its rotational equivalent. Results of tests carried out in novel enclosures, the shapes of which were chosen ad hoc (1) to induce reorientation based only on the ratio of walls lengths plus sense (rectangular enclosure), or (2) to induce reorientation based only on corner angles (rhombus shaped enclosure), suggested that chicks encoded both features of the environment. In a third test, in which chicks faced a conflict between these geometric features (mirror parallelogram-shaped enclosure), reorientation seemed to depend on the salience of corner angles. These results shed light on the elements of the environmental geometry which control spatial reorientation, and broaden the knowledge on the geometric representation of space in animals. PMID- 12884081 TI - Genomic structure and alternative splicing of the insulin receptor tyrosine kinase substrate of 53-kDa protein. AB - Insulin receptor tyrosine kinase substrate of 53-kDa protein (IRSp53) is now known to be a key factor in cytoskeleton reorganization. The human IRSp53 was identified as a binding partner with DRPLA protein, a product of the gene responsible for a neurodegenerative disorder, dentatorubral pallidoluysian atrophy, as well as a binding partner with brain-specific angiogenesis inhibitor 1. Previous studies identified at least four isoforms (L-, M-, S- and T-forms) in human, where 511 amino acid residues from the N-terminus were identical, followed by unique sequences of 9-41 amino acid residues. As each isoform had a distinct function, the unique sequences at the C-terminus had a vital role in its function. Here we report that these isoforms were indeed generated by alternative splicing, which was established by experimental and computational studies on human and rodent genomes. Previous biochemical reports suggested that rodents may lack one of the isoforms (L-form). This study solved this issue, as a nucleotide substitution occurred at a splice donor site followed by a large deletion in the rodent genome compared with human, which made the generation of the L-form impossible. This study also revealed overlapping of the IRSp53 and AATK genes coded for by complementary strands. PMID- 12884082 TI - Identification of a novel 2026G-->C mutation of the MRP2 gene in a Japanese patient with Dubin-Johnson syndrome. AB - Dubin-Johnson syndrome is a recessive inherited disorder with conjugated hyperbilirubinemia caused by a dysfunction of multidrug resistance protein 2 (MRP2) on the canalicular membrane of hepatocytes. A mutational analysis of the MRP2 gene was carried out in three Japanese patients and their family members. In two patients, the homozygous mutations c.1901del67 and c,2272del168 were found. In the third patient, a -24C-->T polymorphism and the two mutations c.1901del67 and 2026G-->C were detected. The 2026G-->C mutation was a novel mutation in exon 16 affecting the conversion of Gly(676) to Arg(676) (G676R) in the MRP2 protein, and was not detected in fifty healthy volunteers. The G676R mutation was located in the Walker A motif of the first nucleotide binding domain in the MRP2 protein, and it was suggested that the mutation induced the dysfunction of the MRP2 protein. It was concluded that the compound heterozygosity of the two mutations of the MRP2 gene in the third patient contributed to the induction of hyperbilirubinemia in this case. PMID- 12884083 TI - Mesh reconstruction preventing sacral herniation. AB - Sacral hernias are uncommon defects developing through the pelvic floor after partial or total sacrectomy. We report on a 29-year-old woman, who has been under our care with a cystic formation after perineosacral resection of a rhabdomyosarcoma and partial sacrectomy. The cystic tumor was resected and a mesh repair performed to prevent sacral herniation. The current literature is summarized; etiology and management recommendations of this rare complication are discussed. PMID- 12884084 TI - New catalyst design for polymerization of norbornene esters by reducing intramolecular interaction. AB - We have used density functional theory to study palladium-based catalysts commonly used for the polymerization of norbornene derivatives with an ester group. Exo-exo, exo-endo, and endo-endo isomers of catalyst complexes were investigated; the endo-endo isomer was the most stable and inactive due to an intramolecular interaction between Pd and O of the carbonyl group. Phosphine groups are effective in minimizing the Pd-O interaction in the endo-endo isomer and P(C6H11)3 was found to be the most efficient reagent. The intramolecular Pd-O interactions were estimated using model complexes, and it was demonstrated that they play a crucial role in stabilizing the endo-endo isomer. PMID- 12884085 TI - Molecular modeling of the rabbit colonic (HKalpha2a) H+, K+ ATPase. AB - A model of the HKalpha2a subunit of the rabbit colonic H+, K+ ATPase has been generated using the crystal structure of the Ca(+2) ATPase as a template. A pairwise sequence alignment of the deduced primary sequences of the two proteins demonstrated that they share 29% amino acid sequence identity and 47% similarity. Using O (version 7) the model of HKalpha2a was constructed by interactively mutating, deleting, and inserting the amino acids that differed between the pairwise sequence alignment of the Ca(+2) ATPase and HKalpha2a. Insertions and deletions in the HKalpha2a sequence occur in apparent extra-membraneous loop regions. The HKalpha2a model was energy minimized and globally refined to a level comparable to that of the Ca(+2) ATPase structure using CNS. The charge distribution over the surface of HKalpha2a was evaluated in GRASP and possible secondary structure elements of HKalpha2a were visualized in BOBSCRIPT. Conservation and placement of residues that may be involved in ouabain binding by the H+, K+ ATPase were considered and a putative location for the beta subunit was postulated within the structure. PMID- 12884086 TI - Mode of action and antifungal properties of two cold-adapted chitinases. AB - The mode of action of two chitinases from the Antarctic Arthrobacter sp. strain TAD20 on N-acetyl-chitooligomers and chitin polymers has been elucidated. Identification of the length of chitin oligomers following enzymatic hydrolysis was verified by using HPLC-based analysis. It was observed that the length of the oligomer is important for enzyme action. The enzymes cannot effectively hydrolyze chitin oligomers with a degree of polymerization lower than four. ArChiA is an endochitinase which hydrolyzes chitin substrates randomly, whereas ArChiB is an exochitinase which degrades chitin chains and N-acetyl-chitooligomers from the nonreducing end, releasing N- N'-diacetyl-chitobiose. ArChiB (100 microg/ml) inhibited spore germination and hyphal elongation of the phytopathogenic fungus Botrytis cinerea by 15% and 30%, respectively. A more pronounced effect was observed with ArChiA (100 microg/ml) resulting in 70% inhibition of spore germination and 60% inhibition of germ tube elongation. A slight additive effect was observed, when the two enzymes were used in combination, only on the inhibition of germ tube elongation. PMID- 12884087 TI - Purification and characterisation of two extremely halotolerant xylanases from a novel halophilic bacterium. AB - The present work reports for the first time the purification and characterisation of two extremely halotolerant endo-xylanases from a novel halophilic bacterium, strain CL8. Purification of the two xylanases, Xyl 1 and 2, was achieved by anion exchange and hydrophobic interaction chromatography. The enzymes had relative molecular masses of 43 kDa and 62 kDa and pI of 5.0 and 3.4 respectively. Stimulation of activity by Ca(2+), Mn(2+), Mg(2+), Ba(2+), Li(2+), NaN(3) and isopropanol was observed. The K(m) and V(max) values determined for Xyl 1 with 4- O-methyl- d-glucuronoxylan are 5 mg/ml and 125,000 nkat/mg respectively. The corresponding values for Xyl 2 were 1 mg/ml and 143,000 nkat/mg protein. Xylobiose and xylotriose were the major end products for both endoxylanases. The xylanases were stable at pH 4-11 showing pH optima around pH 6. Xyl 1 shows maximal activity at 60 degrees C, Xyl 2 at 65 degrees C (at 4 M NaCl). The xylanases showed high temperature stability with half-lives at 60 degrees C of 97 min and 192 min respectively. Both xylanases showed optimal activity at 1 M NaCl, but substantial activity remained for both enzymes at 5 M NaCl. PMID- 12884088 TI - A further investigation of the cytochrome b5-cytochrome c complex. AB - The interaction of reduced rabbit cytochrome b(5) with reduced yeast iso-1 cytochrome c has been studied through the analysis of (1)H-(15)N HSQC spectra, of (15)N longitudinal ( R(1)) and transverse ( R(2)) relaxation rates, and of the solvent exchange rates of protein backbone amides. For the first time, the adduct has been investigated also from the cytochrome c side. The analysis of the NMR data was integrated with docking calculations. The result is that cytochrome b(5) has two negative patches capable of interacting with a single positive surface area of cytochrome c. At low protein concentrations and in equimolar mixture, two different 1:1 adducts are formed. At high concentration and/or with excess cytochrome c, a 2:1 adduct is formed. All the species are in fast exchange on the scale of differences in chemical shift. By comparison with literature data, it appears that the structure of one 1:1 adduct changes with the origin or primary sequence of cytochrome b(5). PMID- 12884089 TI - The cellular distribution and oxidation state of platinum(II) and platinum(IV) antitumour complexes in cancer cells. AB - The cellular distribution of platinum in A2780 ovarian cancer cells treated with cisplatin and platinum(IV) complexes with a range of reduction potentials has been examined using elemental analysis (synchrotron radiation-induced X-ray emission). The cellular distribution of platinum(IV) drugs after 24 h is similar to that of cisplatin, consistent with the majority of administered platinum(IV) drugs being reduced. Micro-X-ray absorption near-edge spectra of cells treated with cisplatin and platinum(IV) complexes confirmed the reduction of platinum(IV) to platinum(II). In cells treated, the most difficult to reduce complex, cis, trans, cis-[PtCl(2)(OH)(2)(NH(3))(2)], platinum(IV) was detected in the cells along with platinum(II). The observations are in accordance with the relative ease of reduction of the platinum(IV) complexes used and support the requirement of reduction for activation of platinum(IV) complexes. PMID- 12884091 TI - Bis-intercalative dinuclear platinum(II) 6-phenyl-2,2'-bipyridine complexes exhibit enhanced DNA affinity but similar cytotoxicity compared to the mononuclear unit. AB - The interactions between a series of platinum complexes, including (pyridyl)(6 phenyl-2,2'-bipyridine)platinum(II) hexafluorophosphate (1), three dinuclear bis[(6-phenyl-2,2'-bipyridine)platinum(II)] complexes (2-4), and (4 aminopyridine)(4,6-diphenyl-2,2'-bipyridine)platinum(II) perchlorate (5) with DNA have been investigated. All Pt(II) complexes, except 5, were demonstrated to be DNA intercalators, based on viscosity measurements. Absorption and fluorescence titration results indicated that the addition of a phenyl ring to the 6-phenyl 2,2'-bipyridine ligand dramatically reduced the DNA binding of the Pt(II) complex 5. The dinuclear complexes 2-4 exhibited multiple binding modes of mono/bisintercalation and groove binding, as revealed by viscosity and fluorescence titration measurements. While complexes 2-4 bound to DNA with significantly enhanced affinities as compared to 1, compounds 1 and 2-4 showed similar IC(50) values against a panel of cancer cell lines. In addition, these complexes showed similar cellular uptakes. The results indicated that the cytotoxicity of these (6-phenyl-2,2'-bipyridine)platinum compounds may not be mediated through DNA binding but may involve interacting mechanisms with cellular components other than DNA. PMID- 12884090 TI - NMR study of manganese(II) binding by a new versatile peroxidase from the white rot fungus Pleurotus eryngii. AB - Nuclear magnetic resonance spectroscopy has been used to characterize the versatile peroxidase from Pleurotus eryngii, both in the resting state and in the cyanide-inhibited form. The assignment of most of the hyperfine-shifted resonances has been achieved by two-dimensional NMR, allowing the comparison of the present system with other ligninolytic peroxidases. This information has enabled a detailed analysis of the interaction of the enzyme with one of its reducing substrates, Mn(II). Furthermore, comparison with the data collected on a mutant in the putative Mn(II) binding site, and an analysis of the enzyme kinetic properties, shed light on the factors affecting the function of this novel peroxidase. PMID- 12884092 TI - Changes in the levels of serum-soluble interleukin-2 receptor after surgical stress. AB - PURPOSE: Surgical stress induces alterations in numerous physiological functions, including the cell-mediated immune response. It is known that interleukin-2 receptor (IL-2R) is released from its specific affinity membrane receptor on activated T lymphocytes and then is detected as a form of the alpha-chain of the IL-2R in the bloodstream. The levels of serum-soluble IL-2R (sIL-2R) reflect the quantity of activated T lymphocytes. This study investigated the changes in the serum sIL-2R levels and the relationship of such changes with other cytokines and the number of lymphocytes after abdominal surgery. METHODS: Twenty-four patients who were scheduled to undergo abdominal operations were enrolled in this study. Blood samples of these cases were collected before surgery, and on postoperative days (POD) 1, 3, 7, and 14. The levels of serum sIL-2R were measured by an enzyme linked immunosorbent assay. RESULTS: The levels of serum sIL-2R achieved the maximal values on POD 1, and gradually decreased until POD 14. The levels of serum sIL-2R on POD 1, 3, and 7 were significantly higher than the preoperative levels. There was a significant and positive correlation between the levels of serum sIL-2R and serum IL-6. There were significant and positive correlations between the levels of sIL-2R and the number of white blood cells and neutrophils. Conversely, there was a significantly negative correlation between the levels of serum sIL-2R and the number of lymphocytes. CONCLUSIONS: As high levels of serum sIL-2R were recognized after abdominal operations, the proliferation of T lymphocytes might still be highly activated in a state of surgical stress, though it is popularly acceptable that surgical stress induces a suppression of cell mediated immunity. PMID- 12884093 TI - Fate of human thyroid tissue autotransplants. AB - PURPOSE: We conducted this study to establish whether human thyroid tissue autografts can survive and function in the absence of their native blood supply in muscle. The benefits of this potential could be incorporated in routine surgery to reduce the incidence of post-operative hypothyroidism. METHODS: Fifteen patients with benign thyroid disorders, seven of whom had Graves' disease and eight, multinodular goiter (MNG), underwent modified subtotal thyroidectomy and the autotransplantation of thyroid tissue in the sternocleidomastoid muscle. About 3-5 g of thyroid tissue was cut and implanted into the sternocleidomastoid muscle. Postoperative clinical assessment, thyroid function tests, and technetium scans of the neck were done to assess the function of remnant and transplanted thyroid tissue. RESULTS: The transplanted tissue was functional in six of the eight patients with MNG and four of the seven with Graves' disease. All the patients with MNG and a functional transplant became euthyroid within 6 months postoperatively. Although the transplanted tissue was functional in four patients with Graves' disease, only one became euthyroid, while the other three required supplemental hormone therapy for postoperative hypothyroidism. CONCLUSIONS: These findings demonstrate the ability of autotransplanted thyroid tissue to survive, function, and grow in muscle. PMID- 12884094 TI - Prediction of 5-fluorouracil and cisplatin synergism for advanced gastrointestinal cancers using a collagen gel droplet embedded culture. AB - PURPOSE: The combination effects of 5-fluorouracil (5-FU) and cisplatin (CDDP) are herein reported using a drug sensitivity assay, with a special focus on the induction of apoptosis. METHODS: The combination effects of 5-FU and CDDP were examined using in vitro chemosensitivity testing by means of the collagen gel droplet embedded culture drug sensitivity test (CD-DST) in MKN45 and GSS cell lines, and primary gastrointestinal cancer cells ob-tained from 40 patients. Apoptosis was assayed using the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate biotin neck end labeling (TUNEL) method. RESULTS: The combination of 5-FU with CDDP increased the efficacy of 5-FU 1.16-1.35-fold with the GSS cell line and 1.10-2.01-fold with primary gastric and colorectal cancers when the exposure time was 7 days. In primary tumor cells a synergistic action was noted in 15 of 40 (38%) gastric and colorectal cancers. Both 5-FU and CDDP were found to induce apoptosis in GSS and MKN45 cells, and the number of apoptotic cells increased synergistically after the combined treatment in the GSS cases and showed a correlation with the results of CD-DST. CONCLUSIONS: Our findings suggested that the efficacy of the combined treatment with 5-FU and CDDP can be predicted by the in vitro chemosensitivity test and that a synergistic effect might be associated with the induction of apoptosis. PMID- 12884095 TI - Topographical electrogastrograms after radical esophagectomy with colonic replacement. AB - PURPOSE: To characterize the functional substitution of colon for the esophagus, we compared the electrogastrogram (EGG) maps and spectral frequencies and power of preoperative controls with patients who had undergone colonic replacement. METHODS: Monopolar EGGs were recorded and spectrally analyzed at 27 locations on the thoracoabdominal surface. The spectral powers of five frequency groups were converted into EGG maps. RESULTS: In contrast to the epigastric concentrations of maximal power foci in a preoperative 3-cpm (cycles per minute) group, those of the colon replacement subjects seemed not to be concentrated in the epigastric region. Power in the 6-cpm colon replacement group were significantly greater and those in the 3-cpm colon replacement group were significantly less than those in the preoperative controls. Spectral frequencies in the 1- and 3-cpm colon replacement groups were significantly higher than those in the preoperative controls. CONCLUSIONS: The colonic and gastric EGG activities had 3-cpm, and probably 6-cpm in common. However, the colonic EGG activities were significantly different from the gastric EGG activities in frequency in the 3-cpm group, and in amplitude in both the 3- and 6-cpm groups. Thus, the replaced colon seems to preserve the original colonic EGG activity. PMID- 12884096 TI - Carcinoembryonic antigen to detect hepatic metastases of colorectal cancers. AB - PURPOSE: The spread of colorectal carcinoma (CRC) affects both staging and treatment. In this prospective study we examined the relationship between carcinoembryonic antigen (CEA) levels in bile, mesenteric venous blood, and peripheral venous blood, and the presence of occult hepatic metastases in patients with CRC. METHODS: Sixty patients who underwent elective surgery at Ankara Oncology Hospital were voluntarily assigned to one of the three groups of 20 patients each. The control group consisted of patients operated on for benign biliary disorders, the second group consisted of patients with CRC without metastases, and the third group consisted of patients with CRC and isolated liver metastases. Bile, mesenteric venous blood, and peripheral venous blood was collected perioperatively for the measurement of CEA levels. RESULTS: The bile CEA levels were significantly higher in the CRC patients with metastases than in those without metastases and the controls. The CEA levels in the mesenteric venous blood and peripheral venous blood were significantly higher in the two groups of CRC patients than in the control group. Overt hepatic metastases later developed in seven patients from the CRC without metastases group who had relatively high CEA levels, but the difference was not significant. CONCLUSION: The CEA levels in bile, mesenteric venous blood, and peripheral venous blood were increased in patients with CRC and liver metastases. The detection of elevated CEA levels in bile taken at surgery did not herald metachronous hepatic metastases in CRC patients without liver metastases. PMID- 12884097 TI - Usefulness of peak-to-peak pulsatility index in infrainguinal bypass graft surveillance. AB - PURPOSE: We conducted this study to find out whether the peak-to-peak pulsatility index (PPI) predicted graft failure and which factors affected the PPI. METHODS: Color-duplex sonography was used to take 520 scans of 74 infrainguinal bypasses, 62 of which were femoropopliteal bypasses and 12 of which were femorocrural bypasses. Vessel diameter and velocity waveform were measured in the graft as well as in the proximal and distal arteries. RESULTS: There were 13 cases of graft failure. The PPI in the mid-graft was significantly different in the normal group (12.30 +/- 8.77) and the graft failure group (4.17 +/- 1.79). A PPI of less than 7.0 in the mid-graft was defined as graft failure, with a sensitivity of 96.0% and a specificity of 77.6%. There was no correlation between the graft diameter and the PPI. The average mode frequency was inversely correlated with the PPI (PPI = 44.8 x Mode F(-1) + 3.50, correlation co-efficient: 0.78). CONCLUSION: One point measurement of the PPI in the mid-graft could be a simple and useful parameter for diagnosing graft failure, based on our finding that the waveform was very similar in the proximal artery, the entire graft, and the distal artery. PPI was inversely correlated with average mode frequency because reverse flow decreased or diminished when a significant stenosis existed. PMID- 12884098 TI - Beneficial effects of n-9 eicosatrienoic acid on experimental bowel lesions. AB - PURPOSE: Dietary fortification of n-9 polyunsaturated fatty acids (PUFA) or 5,8,11-eicosatrienoic acid (ETrA) as well as n-3 PUFA might contribute to the suppression of leukotriene B4 (LTB4) synthesis and thereby reduce inflammatory bowel lesions. As a result, the effect of an ETrA-enriched diet on experimental bowel lesions was examined in this study. METHODS: In Expt. 1, rats were freely fed either an ETrA-enriched or a standard diet. After 7 days of feeding, acute bowel lesions were induced by the subcutaneous injection of 10 mg/kg indomethacin. In Expt. 2, chronic bowel lesions were made by performing subcutaneous injections of 7.5 mg/kg indomethacin twice. After the first injection, the rats were freely fed either an ETrA-enriched or a standard diet for 7 days. RESULTS: In both experiments, the rats fed an ETrA-enriched diet showed increased levels of ETrA in the plasma and intestinal mucosa, and a decreased inflammation score. However, there was no significant decrease in plasma and intestinal mucosal LTB4 in the ETrA-enriched diet-fed rats. CONCLUSION: These results suggest that the dietary supplementation of ETrA may have both prophylactic and therapeutic effects on experimentally produced bowel lesions. Further investigations are necessary to clarify the effects of ETrA on bowel lesions and its mechanisms. PMID- 12884099 TI - Left renal vein anomaly associated with abdominal aortic aneurysm surgery: report of a case. AB - Retroaortic left renal vein occurs infrequently and poses potential hazards to the surgeon during abdominal aortic surgery. We herein report the case of a 77 year-old man with a retroaortic left renal vein who underwent an abdominal aortic aneurysm (AAA) repair. The retroaortic left renal vein was diagnosed preoperatively by contrast-enhanced computed tomography (CT). The patient underwent a successful abdominal aortic replacement with an uneventful postoperative course without any renal complications. The CT scan was useful for both evaluating the AAA and for establishing the presence of the venous anomaly. Major venous anomalies (retroaortic left renal vein, left renal vein collar, left sided inferior vena cava, and caval duplication) should thus be taken into consideration in AAA operations. PMID- 12884100 TI - Simultaneous operations for abdominal aortic aneurysm and liver cancer complicated by severe ischemic heart disease: report of a case. AB - We performed successful simultaneous operations for an abdominal aortic aneurysm (AAA) and liver cancer in a patient complicated by severe ischemic heart disease. A 59-year-old man with a history of liver dysfunction presented with acute epigastric pain. Abdominal computed tomography findings of ascites and a liver tumor indicated a diagnosis of ruptured hepatocellular carcinoma. He had a concomitant 65-mm AAA and a 48-mm right common iliac aneurysm. Elective surgery was scheduled because of his good general condition. Although triple-vessel disease was detected preoperatively, there were no graftable coronary arteries. The aneurysms were repaired first to utilize intra-aortic balloon pumping (IABP) during resection of the liver cancer, followed by left lateral segmentectomy. Perioperative hemodynamics were maintained by administering catecholamines and vasodilators, without the need for IABP. The patient was discharged on the 21st postoperative day without any complications, and no recurrence of liver cancer has been found in the 5 months since his operation. PMID- 12884101 TI - A small bowel volvulus caused by a mesenteric lipoma: report of a case. AB - A 31-year-old man underwent a laparotomy for acute intestinal obstruction symptoms, which he had intermittently experienced for 14 years. The cause of the obstruction was due to a volvulus of the small bowel caused by a mesenteric lipoma. This is a rare finding, which is ideally diagnosed by computed tomography, with surgery the best and most highly recommended treatment. This particular presentation, to the best of our knowledge, has not yet been previously reported in the English language. PMID- 12884102 TI - Enteral migration of a Pezzer tube after a feeding jejunostomy: report of a case. AB - We herein report a rare complication of the migration of a feeding jejunostomy tube through the entire intestine. A surgical Stamm jejunostomy was performed in a patient with an unresectable gastric cancer using a 28-F silicone catheter with a mushroom tip (Pezzer catheter). The catheter was suture-fixed to the skin, family members were given instructions regarding tube feeding and tube care, and the patient was discharged to home care. Two months later, he presented because of the "disappearance" of the tube. A clinical examination revealed a mature jejunostomy tract, skin erosion at the site where the suture was placed, and absence of the tube. There were no signs of intestinal obstruction or peritonitis. Abdominal X-ray examination showed the catheter inside the jejunum. The patient was treated conservatively with serial radiographs showing rapid tube migration through the intestine, and the tube was eliminated spontaneously 5 days later. An awareness of this complication and its appropriate treatment is important given the widespread use of enteral nutrition. PMID- 12884103 TI - Sigmoidoduodenal fistula as a rare complication of colonic carcinoma: report of a case. AB - We report a very unusual case of malignant sigmoidoduodenal communication. To the best of our knowledge, this is the first documentation of this entity in the English language literature. A 76-year-old man presented with weakness, severe weight loss, foul-smelling eructations, anemia, constipation, and episodes of diarrhea. A sigmoidoduodenal fistula was found by barium enema, and a diagnosis of ulcerative colonic adenocarcinoma was made from the colonoscopy findings. Thus, we performed sigmoid colectomy with resection of the fistula and the involved anterior wall of the third duodenal part, followed by primary closure of the duodenal defect. Histological examination confirmed a Dukes' B (Stage II - T(4)N(0)M(0)) colonic adenocarcinoma, and the excision margins of the resected duodenal specimen were clear. We gave adjuvant chemotherapy with 5-fluorouracil and leucovorin. The patient is still alive and disease-free, 2 years postoperatively. PMID- 12884104 TI - Anal endosonography in the diagnosis and management of perianal endometriosis: report of a case. AB - We report a rare case of perianal endometriosis, diagnosed in a 39-year-old woman who presented with a several-day history of a painful mass in the perineum. Perianal examination showed redness and swelling in the right anterior direction. A soft tumor was palpated, but there was no evidence of an episiotomy scar, or of fistula orifices. An anal endosonography in the right anterior direction revealed a sharply defined lesion, 17 x 14 mm in diameter, with high echoic enhancement at its center. The lesion was located along the edge of the external anal sphincter but did not involve it. Based on these endosonographic findings, the tumor was not considered to be an abscess or fistula. We detected its location, and judged it possible to enucleate the tumor under local anesthesia without injuring the anal sphincter. The operation was performed uneventfully and a histological diagnosis of endometriosis was confirmed. Using anal endosonography, we were able to determine the exact anatomic relationship of the lesion in the internal and external sphincter, which substantially influenced the diagnosis and operative procedures. PMID- 12884105 TI - Acute acalculous cholecystitis associated with aortic dissection: report of a case. AB - Acute acalculous cholecystitis is uncommon, but not rare. Classically, this disease is observed in the intensive care unit associated with major trauma, burns, or surgery. Moreover, comorbidity such as infection, hypertension, and diabetes mellitus is often found. Although the exact pathogenesis is still not fully understood, it may be multifactorial and ischemia seems to play a central role. We herein report an unusual case of acute alithiasic cholecystitis predisposing to a de Bakey type III aortic dissection. A 57-year-old man was referred to our hospital for investigation of persistent right upper abdominal pain with tenderness and fever, associated with a newly diagnosed aortic dissection treated conservatively. The diagnosis of acalculous cholecystitis, which is often difficult to establish, was particularly delayed. An open cholecystectomy was performed, revealing a preperforating gangrenous gallbladder without any stones. The patient was discharged from hospital 9 days postoperatively without any early or late complications. No operative treatment for the aortic dissection was needed. PMID- 12884106 TI - Spontaneous splenic rupture after a left-side thoracotomy: report of a case. AB - We herein describe the case of a 59-year-old man who experienced a spontaneous splenic rupture 12 h after undergoing a left-side thoracotomy for a wedge resection of an unknown pulmonary nodular tumor following a history of malignant melanoma. He demonstrated no special abdominal diseases or traumas, except an uneventful cholecystectomy 12 years previously. Preoperatively, he was not on anticoagulation, aspirin, or nonsteroidal anti-inflammatory medication, and all coagulation tests were inconspicuous. At 12 h after lung surgery the patient showed signs of progredient hypovolemic shock. After ultrasonography, which showed a moderate amount of free intra-abdominal liquid, the patient was urgently taken to the operation room. Bleeding resulted from a rupture of an encapsulated hematoma from the spleen. No signs of adhesion around the spleen or of an injury of the left diaphragm were observed. A pathological analysis of the spleen revealed a normal dimension and a normal histological structure without any evidence of a hematological or neoplastic disease. The patient was discharged on the 12th day after surgery. A review on the literature and the differential diagnosis of this unusual case is presented and discussed. PMID- 12884107 TI - Isolated iliac artery aneurysm caused by fibromuscular dysplasia: report of a case. AB - Fibromuscular dysplasia (FMD) can develop in many different arteries, but iliac artery aneurysms are rare. A 69-year-old Japanese woman was admitted to our hospital for treatment of a right common iliac artery aneurysm. Aortography revealed aneurysms in both the right common iliac artery and the left internal iliac artery. Notably, the right common iliac artery aneurysm had a "string-of beads" appearance. At surgery, the aneurysms were resected, and replaced with Y shaped vascular prostheses. The histopathological diagnosis was fibromuscular dysplasia (FMD). We report this case of common iliac artery aneurysm caused by FMD due to its rarity. PMID- 12884108 TI - Closed rectosacropexy for rectal prolapse in children. AB - This report describes a simple, effective, and permanent surgical solution for persistent rectal prolapse (RP). The technique simply involves passing multiple U shaped sutures through the stab incisions made in the skin posterior to the anus, into the sacral fascia, then into the wall of rectum, down to the anal canal, and out through the stab incisions. The strands of the suture are tied subcutaneously through the stab incisions. This operation was successfully performed in 42 children (mean age, 3.5 years) who had suffered from recurrent RP for 3-5 months. None of the children experienced any further recurrence or specific complications during follow-up ranging from 1 to 3 years. Thus, I believe that closed rectosacropexy offers a simple, minimally invasive, and effective method of treating complete RP in children. PMID- 12884109 TI - Signal enhancement through heteronuclear polarisation transfer in in-vivo 31P MR spectroscopy of the human brain. AB - Significant (31)P NMR signal enhancement through heteronuclear polarisation transfer was obtained in model solutions and in vivo on a 1.5-T whole-body MR scanner equipped with two RF channels. The much higher population differences involved in proton Zeeman energy levels can be transferred to the (31)P levels with the refocused INEPT (insensitive nucleus enhancement by polarisation transfer) double-resonance experiment by means of a series of simultaneously applied broadband RF pulses. INEPT achieves a polarisation transfer from (1)H to (31)P spin states by directly reordering the populations in spin systems with heteronuclear scalar coupling. Thus, only the (31)P NMR signal of metabolites with scalar (1)H-(31)P coupling is amplified, while the other metabolite signals in the spectra are suppressed. Compared to Ernst-angle excitation, a repetition time-dependent signal enhancement of eta=(29+/-3)% for methylene diphosphonic acid (MDPA) and eta=(56+/-1)% for phosphorylethanolamine (PE) was obtained on model solutions through optimisation of the temporal parameters of the pulse experiment. The results are in good agreement with numerical calculations of the theoretical model for the studied spin systems. With optimised echo times, in vivo (31)P signal enhancement of the same order was obtained in studies of the human brain. PMID- 12884110 TI - Design of actively shielded main magnets: an improved functional method. AB - An improved functional approach for designing MRI (magnetic resonance imaging) main magnets with active shielding is presented. By nulling one or two external moments as well as a certain series of internal moments of the magnetic field, new designs with improved shielding in combination with or without shorter magnet lengths are obtained. The improved method can be employed to design short and practical superconducting magnets at any given field strength. The resulting designs yield the desired field homogeneity inside the region of interest without using superconducting shim coils. This approach requires only a modest amount of computing power. One of the design steps, a contour plot of the continuous current solutions, can be utilized to study stretch goals for favorable design parameters. PMID- 12884111 TI - Food competition in captive female sooty mangabeys (Cercocebus torquatus atys). AB - We studied the social and foraging behavior of two captive groups of sooty mangabeys under two different spatial food situations. These food conditions were clumped (food was placed in a box) and dispersed (food was dispersed over the entire enclosure). In each group five adult females and two adult males were observed. As a criterion for food competition, individual differences in the relative food intake were used. Adult female mangabeys had a linear, stable, and unidirectional dominance hierarchy. Access to food was rank dependent among females only under clumped food distribution, as current models of the evolution of primate social systems predict. However, feeding success appeared to be mediated not by female but by male agonistic behavior toward females. High ranking females received relatively less aggression from males and could, therefore, stay and feed longer in the feeding area. Male tolerance of higher ranking females seems to mediate female feeding success under restricted food resources. The establishment of a special relationship with a high-ranking male might, therefore, be a strategy to get better access to food. This study demonstrates that female competition for access to food should not be analyzed separately from male influences on females and suggests that a more integral role of males in socioecological models of the evolution of primate social systems should be considered. PMID- 12884112 TI - Changes in unit structures and infanticide observed in Arsi geladas. AB - In 1989 a new gelada baboon ( Theropithecus gelada) population was found in Arsi, on the opposite side of the Rift Valley to that of the known gelada populations of Semien and Showa. Previous comparisons of units of the band at Gado Goro, Arsi, in the same season in consecutive years, indicated that unit structure is less stable among Arsi geladas as compared to the Semien population. Gelada units of the band at Gado-Goro were studied for 7 months in order to investigate the processes of social changes. Changes in unit structure were observed. Provisioning was carried out for 1.5 months at the beginning of the 7-month study period, in order to capture and obtain blood samples from the geladas. Following this, changes in male leadership of some units were observed, presumably as a consequence of the capture. However, natural changes also occurred. One change in unit structure occurred after a female gave birth, and changes in another unit occurred after the disappearance of the leader male. These changes involved female desertion of a unit, her subsequent transfer to a male unit, and culminated in the formation of a unit consisting of one female and one male. One successful and one attempted case of unification of units, and one case of change of a unit leader male are reported. These changes occurred among eight resident units in a period of 7 months (196 female months). Though the types of social changes were not much different from previous observations in Semien National Park, their frequencies seemed to be much higher. The characteristics of Arsi gelada social changes are proposed to be related to the small size of the units. We also describe a new confirmed case and one suspected case of infanticide, as well as one case of abortion at the time of male leader change. PMID- 12884113 TI - Identification of vocalizers by pant hoots, pant grunts and screams in a chimpanzee. AB - Identification of vocalizers was examined using an auditory-visual matching-to sample task with a female chimpanzee. She succeeded in selecting the picture of the vocalizer in response to various types of vocalizations: pant hoots, pant grunts, and screams. When pant hoots by two chimpanzees were presented as a "duet", she could identify both of the vocalizers. These results suggest that researchers have underestimated the capability of vocalizer identification in chimpanzees. The chimpanzee correctly chose her own pictures in response to her vocalizations only by exclusion, and she did not show vocal self-recognition. The effect of acoustical modification (pitch shift and filtration) on the performance suggested that pitch is an important cue for the vocalizer identification. PMID- 12884114 TI - Age-related differences in social grooming among adult female Japanese monkeys ( Macaca fuscata). AB - The present study investigated the influence of dominance rank in combination with kinship on age-related differences in social grooming among adult females in a free-ranging group of Japanese monkeys ( Macaca fuscata). Eighty-three adult females were divided into six sub-groups according to age-class (younger: 5-9 years old; middle: 10-14 years old; older: 15-22 years old) and dominance rank (high and low rank). The ratio of the number of unrelated females that each female groomed to the total number of available unrelated females and grooming bouts which she gave to unrelated females decreased with increasing age for both high- and low-ranking females, whereas age did not appear to affect corresponding values for related females. On the other hand, compared with low-ranking females, high-ranking females of all age-classes received grooming more often from a larger number of unrelated females. Moreover, older females of low rank received grooming less often from a smaller number of unrelated females than younger females of low rank. These results indicate that with increasing age females are more likely to concentrate on related females when they have grooming interactions with other females. This tendency seems to be more apparent for low ranking females. Moreover, the present findings also indicate that older high ranking females could maintain their social attractiveness as high as younger high-ranking females. PMID- 12884115 TI - Cholesterol-fed ovariectomized monkeys are good animal models for human atherosclerosis of postmenopausal women. AB - Although it is well known that the incidence of atherosclerosis is markedly increased in postmenopausal women, antiatherosclerotic effects of estrogen replacement therapies are not clear. One of the reasons for this is due to the lack of appropriate animal models for atherosclerosis of postmenopausal women. Therefore, we attempted to develop an animal model for atherosclerosis of postmenopausal women and examined the antiatherosclerotic effects of estrogen replacement therapy. Adult ovariectomized Japanese monkeys were fed 2% cholesterol diet alone (C-group) or in combination with conjugated estrogen (CE group) for 30 months. The serum estradiol-17beta levels of the CE-group were varied between 10 and 204.5 ng/dl during treatment. In the C-group, the serum total cholesterol levels were increased from 110 to 270 mg/dl, and atheroma was first observed after 3-months treatment with angioscopy. In the CE-group, the levels of the serum total cholesterol during treatment were 30% lower than those of the C-group, and the aortic lesions were first observed after 12-months treatment with angioscopy. The aortic intimal thickness of the CE-group was 58% of the C-group. This finding showed good agreement with the angioscopic observation. The aortic lesions were of a fibromuscular type in both groups. In conclusion, a cholesterol-fed ovariectomized monkey is an appropriate animal model for atherosclerosis of postmenopausal women. Furthermore, angiofiberscopic and histopathological observations suggested that estrogen replacement therapy was valid for atherosclerosis of postmenopausal women. PMID- 12884117 TI - A longitudinal study on hand and wrist skeletal maturation in chimpanzees ( Pan troglodytes), with emphasis on growth in linear dimensions. AB - Skeletal maturation in the chimpanzee hand and wrist (the RUS system; radius, ulna, and short bones) was studied both longitudinally and cross-sectionally. Maturity states were evaluated in each of the 13 bones of the RUS system based on the TW2 method (Tanner and Whitehouse method), and the RUS score was calculated by the summation of scores for these bones. Individual variation was examined by means of residual curves and pseudo-velocity curves of RUS score and anterior trunk length (ATL). Norms of the age change pattern in RUS skeletal maturation and the growth of ATL were determined for each sex, and the relationships among ATL growth and skeletal and reproductive maturation were examined. We found a fairly good relationship between ATL growth and RUS skeletal maturation. Comparison of growth and development between humans and chimpanzees showed that growth characteristics are coupled with each other at puberty in male chimpanzees and in both sexes of humans. Although nutritional condition influenced ATL growth in infancy, it had no effect on the RUS maturational process. Social relationships appeared to influence both ATL growth and RUS maturation. Analyses on relationships between RUS skeletal maturation, ATL growth, and reproductive maturation, showed that RUS skeletal maturation is a good indicator of "physiological age". PMID- 12884116 TI - Inhibin B is the major form of inhibin secreted from testes in male Japanese macaques ( Macaca fuscata). AB - In order to clarify the cellular source and forms of bioactive inhibin in male Japanese macaques ( Macaca fuscata), circulating concentrations of inhibin A and B, and immunohistochemical localization of inhibin subunits in testis were studied. Plasma concentrations of testosterone were also measured. The present study showed that inhibin B was clearly detected in the plasma of male Japanese macaques. Moreover, concentrations of both inhibin B and testosterone during the breeding (mating) season were significantly higher than those of the non-breeding season. On the other hand, plasma inhibin A was detected neither during the breeding seasons nor during the non-breeding seasons. Positive stainings with alpha and betaB subunit antibodies were observed in the Sertoli cells, however staining with betaA subunit antibody was not observed in the testicular samples. These results indicate that inhibin B is the major circulating inhibin and probably secreting from Sertoli cells in male Japanese macaques. PMID- 12884118 TI - Plasma insulin-like growth factor-I, testosterone and morphological changes in the growth of captive agile gibbons ( Hylobates agilis) from birth to adolescence. AB - We examined growth changes in concentrations of plasma insulin-like growth factor 1 (IGF-1) and testosterone, and somatometric parameters in two captive male agile gibbons from birth to about 4 years of age, to examine the evolution of growth patterns in primates. Plasma IGF-1 concentrations in agile gibbons generally increased with age with values ranging from 200 to 1100 ng/ml. The growth profiles in plasma IGF-1 in the gibbons were similar to those reported for chimpanzees. The highest concentrations of plasma testosterone (230 and 296 ng/dl) were observed within the first 0.3 years from birth, then the concentrations rapidly decreased and fluctuated below 100 ng/dl. Continuously higher IGF-1 concentrations were observed after 2.6 and 3.5 years of age. The profiles of plasma testosterone in these gibbons also resembled those of other primates including humans. However, their plasma testosterone levels in both neonate and adult stages (60 ng/dl) were lower than those reported for macaques and chimpanzees of respective stages. The obtained growth profiles of plasma IGF 1 and testosterone suggest that the adolescent phase starts around 2.6 or 3.5 years of age in male agile gibbons. The growth trend in many morphological parameters including body weight showed a linear increase without a significant growth spurt at approximately the onset of puberty. Head length and first digit length had reached a plateau during the study period. Brachial index, which indicates the relative length of forearm to upper arm, significantly increased gradually through the growth period. This result indicates that forearm becomes relatively longer than the upper arm with growth, which may be an evolutionary adaptation for brachiation. PMID- 12884119 TI - Familial early onset macular degeneration in cynomolgus monkeys (Macaca fascicularis). AB - The mode of inheritance of macular degeneration was determined with 45 cynomolgus monkeys (18 females and 27 males) who were the offspring of one breeding male with typical macular degeneration. In the first generation, 27 offspring (10 females and 17 males) were born from mating between the macular degeneration affected founder male and 5 normal female breeders. Among them, 18 monkeys (9 females and 9 males) were judged as having macular degeneration (affected). Next, the distribution of affected offspring was examined with 18 offspring who were born from 3 different mating pairs, normal vs normal, affected vs normal and affected vs affected, when they became 2 years old. All of the 9 monkeys (4 females and 5 males) obtained from the 2 pairs of normal vs normal were normal. On the other hand, 6 affected monkeys (3 females and 3 males) were detected in 8 offspring from the mating pair of affected vs normal, and the single offspring produced by the mating pair of affected vs affected was affected. These results showed that this degeneration must be early onset familial macular degeneration controlled by autosomal dominant gene(s). PMID- 12884125 TI - Long-term adaptation of methanol-fed thermophilic (55 degrees C) sulfate-reducing reactors to NaCl. AB - A laboratory-scale upflow anaerobic sludge bed (UASB) reactor was operated during 273 days at increasing NaCl concentrations (0.5-12.5 g NaCl l(-1)) to assess whether the stepwise addition of the salt NaCl results in the acclimation of that sludge. The 6.5-l thermophilic (55 degrees C), sulfidogenic [a chemical oxygen demand (COD) to SO4(2-) ratio of 0.5] UASB reactor operated at an organic loading rate of 5 g COD l(-1) day(-1), a hydraulic retention time of 10 h and was fed with methanol as the sole electron donor. The results show that the adaptation of the thermophilic, sulfidogenic methanol-degrading biomass to a high osmolarity environment is unlikely to occur. Sulfide was the main mineralization product from methanol degradation, regardless of the NaCl concentration added to the influent. However, sulfide production in the reactor steadily decreased after the addition of 7.5 g NaCl l(-1), whereas acetate production was stimulated at that influent NaCl concentration. Batch tests performed with sludge harvested from the UASB reactor when operating at different influent salinities confirmed that acetate is the main metabolic product at NaCl concentrations higher than 12.5 g l(-1). The apparent order of NaCl toxicity towards the different trophic groups was found to be: sulfate-reducing bacteria > methane-producing archaea > acetogenic bacteria. PMID- 12884126 TI - Biofilms, bacterial signaling, and their ties to marine biology. AB - Much of what is know about quorum sensing has come from the study of marine biology. The original description of the phenomenon was based on the study of marine bacteria and the luminescent pathway. More recently, aquatic organisms have been found to inhibit bacterial fouling of surfaces by blocking signaling pathways in the bacteria. These signaling effects have, over the last 5 years, been linked to biofilms. However, this correlation is not as straight forward as originally believed. Here, a brief overview of quorum sensing, and background on biofilms is provided, followed by a discussion of more recent work looking at the effects that environment may have on signal expression. PMID- 12884127 TI - AfsR as an integrator of signals that are sensed by multiple serine/threonine kinases in Streptomyces coelicolor A3(2). AB - The genome sequence of Streptomyces coelicolor A3(2) has revealed the presence of about 40 protein serine/threonine or tyrosine kinases. AfsK, which is able to phosphorylate AfsR, a transcriptional activator with ATPase activity, represents the first instance in which a bacterial Hanks-type protein kinase phosphorylates a specific protein and exerts biologically important functions. The AfsK-AfsR system in S. coelicolor A3(2) globally controls secondary metabolism. The signal transduction pathway so far demonstrated or suggested is as follows: AfsK loosely attached to the membrane autophosphorylates threonine and serine residues, perhaps on sensing some external stimulus, and enhances its kinase activity. The kinase activity is modulated by KbpA, an AfsK-binding protein, by means of protein-protein interactions. The activated AfsK phosphorylates threonine and serine residues of AfsR in the cytoplasm, by which the DNA-binding activity of AfsR is greatly enhanced. In addition to AfsK, other kinases-including PkaG and AfsL-also phosphorylate AfsR, suggesting that AfsR serves as an integrator of multiple signals sensed by these kinases. The phosphorylated AfsR binds the promoter of afsS, which encodes a protein of 63 amino acids, and forms a closed complex with RNA polymerase. The closed complex is then converted to a transcriptionally active open complex by the energy available from ATP hydrolysis by AfsR. AfsS induced in this way activates transcription of pathway-specific transcriptional activators, such as actII-ORF4 for actinorhodin production and redD for undecylprodigiosin, in an as yet unknown manner. PMID- 12884128 TI - Antibiotic activity of lectins from marine algae against marine vibrios. AB - Saline and aqueous ethanol extracts of marine algae and the lectins from two red algal species were assayed for their antibiotic activity against marine vibrios. Experimental studies were also carried out on the influence of environmental factors on such activity, using batch cultures. The results indicated that many of the saline extracts of the algal species were active and that the activity was selective against those vibrios assayed. The algal extracts were active against Vibrio pelagius and the fish pathogen V. vulnificus, but inactive against V. neresis. Algal lectins from Eucheuma serra (ESA) and Galaxaura marginata (GMA) strongly inhibited V. vulnificus but were inactive against the other two vibrios. The antibacterial activity of algal extracts was inhibited by pretreatment with various sugars and glycoprotein. Extracts of the two red algae, E. serra and Pterocladia capillacea, in saline and aqueous ethanol, inhibited markedly the growth rate of V. vulnificus at very low concentrations. Culture results indicated that metabolites active against V. vulnificus were invariably produced in P. capillacea over a wide range of temperature, light intensity, and nutritional conditions. Enhanced antibacterial activity occurred when P. capillacea was grown under higher irradiance, severe nutrient stress and moderate temperature (20 degrees C), reflecting the specific antibiotic characteristics of this alga. The strong antibiotic activity of lectins towards fish pathogenic bacteria reveals one of the important roles played by algal lectins, as well as the potential high economic value of those marine algae assayed for aquaculture and for biomedical purposes. PMID- 12884129 TI - Phylogeny of Saururaceae based on mitochondrial matR gene sequence data. AB - DNA sequences of matR gene from three species of Saururaceae and the selected outgroups, Chloranthus holostegius and Zippelia begoniaefolia, are reported. All DNA sequences of six species in four genera of Saururaceae and the two outgroups are analyzed on PAUP 4.0 8b to reconstruct the phylogeny. A single matR gene tree is generated from parsimony, distance, and likelihood analyses, respectively. The three trees with the same topology are slightly different in bootstrapping support for some clades. The result indicates that Saururaceae is monophyletic. Anemopsis is sister to Houttuynia, and the two genera form the first diverging lineage of the family. The sister group relationship between Saururus and Gymnotheca is also supported by a relatively high bootstrap value. The result is different from all the former phylogenetic opinions on Saururaceae based on morphology, but it is supported by the evolution of flower-bract stalk in Saururaceae. In addition, some characteristics of the matR gene are analyzed. The MatR gene is a relatively better tool to reconstruct the molecular clock because the base substitution bias greatly decreases in the gene. PMID- 12884130 TI - Identification of mono-, oligo-, and polysaccharides secreted from soybean roots. AB - The mist culture system was conducted to study secreted polysaccharides from soybean ( Glycine max) roots grown for 15 days. Roots were rinsed with distilled water (DW) for 15 min, then with 30 mM oxalic acid (OXA) for 15 min to remove ionically bound sugar. Released sugars were further fractionated into low (L) and high (H) molecular weight fractions with Sephadex G-10. DW rinsing released 190 microg neutral sugar (NS) and 62 microg uronic acid (UA) per plant, while 374 microg NS and 70 microg UA per plant were released by OXA rinsing. Acetylation analysis revealed that the L fraction by DW and OXA mainly consisted of glucose (Glc), pinitol, and UA, whereas the H fraction mainly consisted of arabinose (Ara), galactose (Gal), Glc, and UA. The presence of rhamnose (2%-6%) in both fractions suggests secretion of rhamnogalacturonans. Methylation analysis revealed that the H fraction by DW and OXA contained T-Ara, 3-, 6-, and 3,6-Gal, suggesting the presence of type II arabinogalactan and arabinogalactan proteins. HPLC analysis detected mono-, di-, and tri-GalA in the L fraction by DW and OXA. Substances corresponding to sucrose, kojibiose, cello- and laminari oligosaccharides were also found in root exudates. PMID- 12884131 TI - Cryptic species in the fern Ceratopteris thalictroides (L.) Brongn. (Parkeriaceae). I. Molecular analyses and crossing tests. AB - For the taxonomic revision of the problematic species Ceratopteris thalictroides, molecular analyses and crossing tests were conducted for 16 sources in the world. An analysis of allozyme composition of five enzymes revealed the presence of three intraspecific entities, which were called the south type, the north type, and the third type. An analysis of the nucleotide sequences of chloroplast DNA also distinguished the same entities. Crossing tests showed that the south type was completely cross-sterile with the other two types, and that the other two were considerably cross-sterile with each other. These results suggest that the three entities should be regarded as different biological species. Although the south type and the other two meet in several regions, complete cross-sterility between them seems to sustain their genetic distinctiveness in spite of occasional crossing. The results from the present study suggest that widely distributed fern species are apt to comprise several cryptic species. PMID- 12884132 TI - Predicting the impact of climatic warming on the carbon balance of the moss Sanionia uncinata on a maritime Antarctic island. AB - The effects of climatic factors, especially those of temperature, on the carbon balance of the moss Sanionia uncinata were examined on King George Island in the maritime Antarctic. Net photosynthesis (P(n)) and dark respiration rates of two colonies (A and B) were measured with a portable infrared gas analyzer. Colony A showed small P(n) compared with its dark respiration rates throughout the growing season. Colony B showed much higher net photosynthetic rates, but the dark respiration rates of the two colonies did not differ significantly. Net photosynthetic rate determined at light saturation was almost constant over a wide temperature range, from 5 degrees to 15 degrees C, while the dark respiration was strongly affected by temperature. To assess the impact of warming on the carbon balance of the moss, cumulative carbon gain of the moss was calculated using a simulation model for the main part of the growing season. The results suggest that climatic warming may cause a reduction of carbon gain in some relatively photosynthetically inactive moss colonies. PMID- 12884133 TI - Examination of morphological changes in the first formed protoxylem in Arabidopsis seedlings. AB - We examined morphological changes in the first-formed protoxylem vessels in Arabidopsis seedlings. Between 2.5 and 8 days after imbibition, mean hypocotyl and root length increased 1.52 and 23.3 times, respectively. In the 2.5-day-old seedlings, two continuous protoxylem vessels were present in the hypocotyl-root axis. In the 8-day-old upper hypocotyls, six protoxylem vessels were observed, and in the lower hypocotyls, four protoxylem vessels and one or two metaxylem vessels were observed. In the 8-day-old roots, there were two protoxylem vessels and two or three metaxylem vessels. Two protoxylem vessels in the hypocotyls connected to two metaxylem vessels in the roots of 8-day-old seedlings. At the 0.3-mm part below the hypocotyl-root boundary, the mean intervals of neighboring annular secondary wall thickenings in protoxylem vessels in 8-day-old roots were 12.9% larger than those in 2.5-day-old roots. In more apical parts of 8-day-old roots, the mean intervals fluctuated between 1.71 and 2.29 microm. In 8-day-old seedlings, metaxylem vessels were formed between 0.4 mm above the hypocotyl-root boundary and 17 mm below the boundary. The intervals in these regions were not extended so much as protoxylem vessels were collapsed. The first-formed protoxylem vessels presumably retain their water-conductive function after metaxylem formation. PMID- 12884134 TI - Clonal integration and its ecological significance in Hedysarum laeve, a rhizomatous shrub in Mu Us Sandland. AB - Hedysarum laeve, a rhizomatous clonal half-shrub, commonly dominates in inland dunes in semiarid areas of northern China. This species propagates vegetatively by the extension of horizontal rhizomes resulting in programmed reiteration of apical and/or axillary meristems. In this study, (14)C labeling and experimental defoliation were employed to test the photosynthate translocation within the interconnected parent-daughter ramet pairs. A proportion of (14)C-photosynthates was transported from the parent ramet into the daughter ramet, the roots of the daughter ramet, and the rhizome; these three components showed more than 70% sink activity after 24-h translocation. On the other hand, the basipetal translocation (from daughter ramet into parent ramet) was relatively small with sink activity of less than 5%, but sink activity of the rhizome exceeded 10%. Defoliation had an influence on the photosynthate translocation between parent and daughter ramets. The intact parent ramets significantly increased their (14)C photosynthate translocation into defoliated daughter ramets when compared to intact daughter ramets. The daughter ramets transported significantly more (14)C photosynthates to the defoliated parent ramets than to the intact parent ramets. A portion of (14)C-photosynthates was transported into the rhizome from both parent and daughter ramets, indicating that the rhizome is supported by both ramets for photosynthates. The clonal integration between ramets of the species through rhizome connection may confer benefit both to the ramets and the genet in adverse environments. PMID- 12884135 TI - Analysis of salt-inducible genes in barley roots by differential display. AB - To obtain insight into the comprehensive molecular characteristics related to the mechanisms of salt tolerance, we performed a large-scale screening of salt inducible genes in barley roots by differential display. A comparative analysis of gene expression between control and salt-stressed conditions led to the detection of 218 cDNA clones induced by salt. Sequence analysis and database searching revealed that 133 cDNA clones have homology to known proteins. Twenty four salt-inducible clones were identified as genes for signal transduction (e.g., phosphatidylinositol-4-phosphate-5-kinase, mitogen-activated protein kinase, transcription factor, receptor protein kinase, and protein phosphatase 2A). We also detected clones encoding glutathione reductase, thioredoxin-like protein, trehalose-6-phosphate synthetase, and heat shock proteins in the category of typical stress tolerance. Furthermore, we have obtained genes encoding membrane transporters, members of the P450 family, enzymes involved in RNA metabolism or function, and enzymes of sugar or amino acid metabolism. It must be noted that most genes were expressed strongly in roots, but only rarely or weakly in leaves. In addition, some clones were newly found as salt-inducible genes encoding SCARECROW, splicing factor and apoptosis protein. In this research, it was shown that differential display is a powerful tool for a large scale cloning of cDNAs induced by salt and these results are very useful for understanding the mechanisms of plant salt tolerance. PMID- 12884136 TI - Photogravitropic equilibrium in Avena coleoptiles: fluence rate-response relationships and dependence on dark adaptation and clinostating. AB - Phototropism of Avena coleoptiles was measured in response to blue-light irradiation lasting between 2 and 24 h. During this time the coleoptiles established a bending angle of photogravitropic equilibrium that was dependent on the time of irradiation and also on the pretreatment in light or darkness prior to stimulation. The absolute threshold for the photogravitropic equilibrium in response to blue light was 10(-8) micromol m(-2) s(-1). Photon fluence rate response curves, which were generated after several hours of dark adaptation, had a characteristic shape with a prominent optimum in the middle of the dynamic range. Curves which were generated without prior dark adaptation displayed no such optimum. Clinostating dark-adapted coleoptiles caused an increase of sensitivity and responsiveness during a 2-h period of unilateral irradiation. The advantages and the drawbacks of long-term irradiation experiments for the investigation of phototropism and the generation of action spectra are discussed. PMID- 12884137 TI - Programmed cell death of Pinus nucellus in response to pollen tube penetration. AB - In ovules of Pinus densiflora, pollen tubes elongate and branch into the nucellar tissue in the direction of the female gametophyte. After pollination, nucellar cells located around the pollen grain and tube die off. We showed here that the nuclei of the nucellar cells were stained by TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP-fluorescein nick end labeling). The number of TUNEL positive cells increased during pollen tube growth. The tips of pollen tube branches protruded into the nucellar cells to form a convex-concave junction. At this junction, the cell membrane of nucellar cells was separated from the cell wall and the protoplast shrank. Small vesicles and amorphous materials were released from the protoplast into the space between the cell membrane and wall. Vacuoles were collapsed, chromatin was condensed, and mitochondria and plastids were deteriorated in the shrunken protoplast. Agarose gel analysis of DNA isolated from the ovules showed a DNA ladder, suggesting that the nuclear DNA had undergone internucleosomal cleavage. These results suggest that nucellar cells undergo programmed cell death in response to pollen tube penetration with some features resembling apoptosis and other features peculiar to nucellar cells. PMID- 12884138 TI - The synergid cell: attractor and acceptor of the pollen tube for double fertilization. AB - In flowering plants, the egg cell is generally accompanied by two symmetrical cells, called synergid cells. As early as the 1870s, synergid cells were distinguished from egg cells and cooperation between synergid and egg cells was proposed; the term "synergid" is derived from the Greek "synergos," which means "working together." The accumulation of morphological and genetic data, and, more recently, the in vitro physiological analysis of the fertilization system of Torenia fournieri, have revealed that synergid cells work together with egg and central cells to accomplish double fertilization. This cooperation is of crucial importance in the attraction and acceptance of the pollen tube. In this review article, I focus on the physiological function and behavior of the synergid cell during the fertilization process. PMID- 12884139 TI - [Results of the Magdeburg consensus conference on surgical hernia repair]. PMID- 12884140 TI - [Registration using three-dimensional laser surface scanning for navigation in oral and craniomaxillofacial surgery]. AB - INTRODUCTION: Recent innovations in laser scanning technology provide a potentially useful tool for three-dimensional surface registration for image guided surgery. The purpose of this study is to evaluate the clinical reliability of this technique in oral and maxillofacial surgical procedures using image guided navigation. METHODS: In an experimental step, a stable anthropomorphic skull model with prelabeled markers was scanned and registered with laser surface scanning (z-touch, BrainLAB) and marker- based algorithms. The registration protocol was then repeated 25-times. Root mean square error (RMSE) and target difference values were compared for their suitability for this application. Twelve patients with different indications for oral and maxillofacial surgery were planned for image-guided surgery using a passive infrared surgical navigation system (VectorVision, BrainLAB). Preoperative computed tomography (CT) scans were carried out with newest 16-line multisclice CT-scanner (Siemens Somatom Sensation 16). The new markerless laser surface scanning technique was applied in all intraoperative patient registrations. Registration error was noted. The clinical application accuracy was determined for anatomical landmark localization deviation. RESULTS: In the experimental protocol a mean registration error (RMSE) or target difference of 1.3 (0.14) or 2.08 (0.49) mm for laser scanning and 0.38 (0.01) or 0.99 (0.15) mm for marker registration was found. The differences for RMSE and target localization were statistically significant (p < 0.005). Furthermore, a strong correlation between RMSE and target difference was found for laser scanning (r = 0.96) and marker registration (r = 0.95). During various clinical procedures involving oral and maxillofacial surgery, the overall error of the registration procedure determined as RMSE was 1.21 (0.34) mm. Intraoperatively, the mean clinical application accuracy was found to be 1.8 (0.5) mm. CONCLUSION: Three-dimensional laser surface scanning technique may be a interesting and useful approach to register the patient for image-guided procedures, particularly during oral and craniomaxillofacial surgery. PMID- 12884141 TI - [Rupture of abdominal aortic aneurysm (RAAA)--predictors of the early postoperative mortality]. AB - PURPOSE: To evaluate the main factors of the 30 days mortality rate of patients operated on for abdominal aortic aneurysm rupture (RAAA. PATIENTS AND METHOD: Univariate and multivariate analysis of various factors associated with RAAA was performed in a group of 73 patients operated on for RAAA between 1996-2001. RESULTS: The 30 days mortality rate was 35.6 %. The main factors of mortality were: misdiagnosis, cardio- pulmonary-cerebral resuscitation (CPCR) on admission, configuration of RAAA, number of blood transfusions, hypotension on admission (p < 0.0001) and duration of operation, type of reconstruction and hypertension in anamnesis (p < 0.01). Important factors (p < 0.05) of postoperative mortality were also low haemoglobin level on admission, abdominal aortic aneurysm (AAA) diameter and ischaemic heart disease in anamnesis. The probability of patient's death is the highest (p < 0.003), if factors like CPCR, number of blood transfusions and aneurysm diameter are combined (multivariate analysis, stepwise method). CONCLUSION: The early detection and surgical or endovascular elective treatment of AAA, the regular dispensation of patients with small AAA especially in hypertonics, the correct diagnosis of RAAA without time delay are the best tools for patients survival. The patient's chance for survival increases with highly trained prehospital resuscitation system and experienced team of vascular surgeons and anesthesiologists. PMID- 12884142 TI - [Minimal invasive surgery for pHPT--which patients will profit?]. AB - BACKGROUND: Standard for treatment of pHPT has been the exploration of the neck and extirpation of the macroscopic identified adenoma for a long time. Since the Quick-PTH-Assay is available as intraoperative control of success, it is possible to extirpate a preoperativeley localized adenoma without identifying the normal parathyroid glands. Since the accuracy of the preoperative localization is essential for a minimally invasive or endoscopic procedure, we analyzed retrospectively the accuracy of ultrasound and mibiscan of our patients, operated due to pHPT in our facility. METHODS: Between 1995 and 2002 119 patients were operated due to HPT in our facility. Sensitivity, specificity, positive- and negative-predictive value for diagnosis and correct localisation of a singular adenoma by ultrasound and/or mibiscan were determined. RESULTS: We found a singular adenoma in 94 patients, a carcinoma in 1 patient, 6-times a double adenoma and in 18 cases a hyperplasia of all parathyroid glands. Sensitivity, specificity, positive- and negative-predictive value for diagnosis and correct localisation of a singular adenoma were 52 %, 41 %, 60 % and 27 % rsp., when the results of mibiscan and ultrasound were used. DISCUSSION: Related to the preoperative imaging, a single gland exploration and extirpation would have been possible in 70 % of our patients with pHPT. However, in 40 % of these patients the 100 % reliable Quick-PTH-Assay would have shown, that the adenoma has not been removed successfully in the first attempt. PMID- 12884143 TI - [Minimally invasive options for gastric tumors]. AB - INTRODUCTION: Depending on the origin of the tumor tissue, gastric tumors may be more or less accessible for biopsy diagnostics. However, especially submucous tumors present a diagnostic problem. Entity and dignity may remain unclear particularly in larger tumors and may not be clarified before operative excision via gastrotomy and frozen section diagnostics. Similar problems may occur in the diagnostics of epithelial tumors, if a reliable appraisal of the dignity based on forceps biopsy is impossible. To clarify their entity and dignity, tumors can be completely extirpated with minimally invasive techniques. PATIENTS AND METHODS: Apart from the endoscopic mucosa resection (EMR), laparoscopic intragastric tumor resection and laparoscopic wedge resection were performed, especially in larger tumors. In the period from December 1999 to December 2001, we saw an indication for minimally invasive procedures in 22 patients. There were 5 cases of submucous tumors of unclear entity and 17 epithelial lesions. The epithelial lesions included 12 patients with tumors of unclear dignity and five cases with early gastric carcinomas. RESULTS: The EMR was performed without complications in all 14 cases. One of the three cases with wedge resection was followed by a gastrectomy for oncological reasons. One early postoperative bleeding occurred, which was controlled laparoscopically. Conversion to open surgery due to technical problems was necessary in two cases of laparoscopic intragastric resection, and in one case a gastrectomy was required for oncological intention. CONCLUSION: Beside the diagnostic aspect, the mentioned techniques also enable a minimally invasive therapy of locally excisable gastric tumors. In addition to benign and low grade malignant lesions, early gastric carcinomas of the intestinal type present an indication. PMID- 12884144 TI - [Right heart failure following pericarditis constrictiva tuberculosa--urgent surgical treatment]. AB - A 71-year-old male patient presented with a 4-month history of fever, dyspnoea, night sweat, ankle swelling and was admitted to our institution for further investigation due to heart failure (NHYA IV). A posterior-anterior chest radiograph showed an enlarged cardiac silhouette, the lung was without pathological findings; calcifications were not described. Echocardiography revealed a severe diastolic malfunction but no pericardial effusion. In computed tomography, pericardium was thickened. Patient was admitted for further investigations. Heart catheterization revealed a left ventricular ejection fraction of 56 %, a cardiac index of 1.3 ml/min/m2 leading to the diagnosis of severe constrictive pericarditis. The patient underwent an urgent pericardectomy via median sternotomy. Extracorporal circulation was not necessary. The postoperative course was uneventful, heart failure improved to NYHA II. The removed pericardium revealed severe granulomatous pericarditis resulting from infection with acid-resistant bacilli. The diagnosis was confirmed by a positive culture for mycobacterium tuberculosis. The patient was put on anti-TB chemotherapy for one year. 1 year after operation patient is graduated in NYHA class II. CONCLUSION: This rare extrapulmonary form of TB can have an insidious or sudden onset. The diagnosis is complicated by non-specific clinical and radiographic findings. Clinical presentation may be the result of the infectious process itself or the pericardial inflammation causing pain, effusion, and hemodynamic effects. In the absence of concurrent extracardiac TB, diagnosis of pericardial TB is difficult. Nevertheless, rapid diagnosis and treatment are crucial to reduce mortality. PMID- 12884145 TI - [Endosonographically controlled transluminal fine needle aspiration biopsy: diagnostic quality by cytologic and histopathologic classification]. AB - BACKGROUND AND OBJECTIVE: EUS-guided fine needle aspiration (EUS-FNA) has emerged as a highly accurate technique for detecting and classifying mediastinal and pancreatic lesions as well as abdominal and recently retroperitoneal masses with a minimum of risk for the patient. PATIENTS AND METHODS: To objectify these statements, we evaluated the quality of 72 EUS-FNA specimens by cytologic and histopathologic classification, investigated their contamination with tissue from the needle pathway and observed puncture-related complications in a retrospective study of 44 EUS-FNA in 41 consecutive patients (56 +/- 14 years, m = 24, f = 17; 13 pancreatic, 9 adrenal, 6 abdominal and 13 mediastinal masses). EUS-FNA was performed using a PENTAX 32 UA endosonoscope (longitudinal 7.5 MHz sector array) in combination with a needle system type "Hancke-Vilmann". RESULTS: 16 vs. 11 of 34 histopathologic and 38 cytologic specimens were classified "excellent", 7 vs. 10 "sufficient", 7 vs. 13 "poor" and 4 vs. 4 "failed". Analysis of contamination with tissue from the needle pathway showed 4 vs. 2 specimens "highly", 3 vs. 14 "clearly", 8 vs. 19 "slightly" and 19 vs. 3 "not" contaminated. Specimens classified "excellent" were less contaminated (p = 0,037). EUS-FNA identified 35 benign and 24 malignant masses. Definite diagnosis failed in 13 specimens. One nonfatal complication occurred. EUS-FNA is an accurate (89 %) and low-risk procedure to examine primary undiagnosed mediastinal, pancreatic, intraabdominal and especially adrenal lesions in most of the cases. Contamination with tissue from the needle pathway seems to be a major predictive factor of poor specimen quality and failed diagnosis. CONCLUSION: EUS-FNA expands the diagnostic approach of mediastinal, abdominal, pancreatic and adrenal masses and provides accurate specimens for reaching new differential-diagnostic competence, especially in endocrinologic cases. PMID- 12884146 TI - [Non-healing gastric ulcer]. AB - HISTORY AND ADMISSION FINDINGS: A 47-year-old Angolan complained of upper abdominal pain, fever, weight loss and night sweats. Abdominal ultrasound performed by his general practitioner showed enlarged lymph nodes at the liver hilum. Physical examination was negative except for slight tenderness over the right upper abdomen. INVESTIGATIONS: A tuberculin skin test was clearly positive. Upper gastrointestinal endoscopy revealed multiple gastric ulcers and the Helicobacter pylori test was positive. The lymph nodes at the liver hilum were needle-aspirated, this showed granulomatous inflammation similar to the biopsies of the ulcers. Eradication of Helicobacter pylori and therapy with proton pump inhibitors were initiated. DIAGNOSIS, TREATMENT AND COURSE: Epigastric pain and the ulcers persisted unchanged. The culture of the lymph node aspirate grew multiresistent Mycobacterium tuberculosis. With adequate tuberculostatic therapy the patient improved rapidly and the further course was without any complications. CONCLUSION: Tuberculosis should be included in the differential diagnosis in non-healing ulcers of the gastrointestinal tract, especially in immigrants from endemic areas. PMID- 12884147 TI - [Tolerance induction in anaphylaxis to carboplatin]. PMID- 12884148 TI - [Surgical options in chronic pancreatitis]. PMID- 12884149 TI - [New data on the pathogenesis of steroid-induced osteoporosis: consequences for therapy and prevention]. PMID- 12884150 TI - [Genetics of osteoporosis]. PMID- 12884151 TI - [Evidence-based clinical practice guidelines for diagnosis and treatment of osteoporosis]. PMID- 12884152 TI - [Is lifetime anticoagulation required in factor V Leiden mutation?]. PMID- 12884153 TI - [Evidence-based therapy of osteoporosis--comment on DMW 17/2003]. PMID- 12884154 TI - Association between acquisition of herpes simplex virus type 2 in women and bacterial vaginosis. AB - A longitudinal cohort study of sexually active women 18-30 years of age was conducted to identify variables associated with the acquisition of herpes simplex virus type 2 (HSV-2) infections. Six hundred seventy HSV-2-seronegative women were followed up at 4-month intervals for 1 year; acquisition of HSV-2 antibodies was detected in 32 of these women. Black race, < or =12 years of education, having a new sex partner, and bacterial vaginosis (BV) were associated with HSV-2 seroconversion on univariate analysis. Antecedent HSV-1 infection was not protective against HSV-2 acquisition. After controlling for other identified risk factors in multivariable models, the diagnosis of BV remained associated with an increased risk of acquiring HSV-2 infection (hazard ratio, 2.1; 95% confidence interval, 1.0-4.5; P=.05). In this study, the population attributable risk of BV for HSV-2 seroconversion was 21%. Additional studies are needed to determine whether screening and treatment of BV could reduce susceptibility to the acquisition of HSV-2 in women. PMID- 12884155 TI - Control of endemic vancomycin-resistant Enterococcus among inpatients at a university hospital. AB - We sought to determine the ability of surveillance cultures and isolation of vancomycin-resistant Enterococcus (VRE)-colonized patients to control nosocomial VRE infection and colonization during a 5-year period (November 1994 through October 1999). During this period, VRE colonization was limited to 0.82% of admissions. The incidence of VRE infection was 0.12 cases per 1000 patient-days (attack rate, 0.07%). Colonized patients were first identified by surveillance (95%) or routine clinical cultures (5%); 14% of colonized patients had a positive clinical culture a median of 15 days after a positive surveillance culture. Ten percent of colonized patients were identified by surveillance at the time of transfer from another health care facility. Identification of these colonized patients was associated with reduction from a peak incidence rate of 2.07% to a rate of 1.25% and stabilization at this lower level. The use of surveillance cultures to identify and isolate patients with asymptomatic colonization can provide sustained control of the spread of VRE within a health care facility. PMID- 12884156 TI - Intravenous immunoglobulin G therapy in streptococcal toxic shock syndrome: a European randomized, double-blind, placebo-controlled trial. AB - The efficacy and safety of high-dose intravenous polyspecific immunoglobulin G (IVIG) as adjunctive therapy in streptococcal toxic shock syndrome (STSS) were evaluated in a multicenter, randomized, double-blind, placebo-controlled trial. The trial was prematurely terminated because of slow patient recruitment, and results were obtained from 21 enrolled patients (10 IVIG recipients and 11 placebo recipients). The primary end point was mortality at 28 days, and a 3.6 fold higher mortality rate was found in the placebo group. A significant decrease in the sepsis-related organ failure assessment score at days 2 (P=.02) and 3 (P=.04) was noted in the IVIG group. Furthermore, a significant increase in plasma neutralizing activity against superantigens expressed by autologous isolates was noted in the IVIG group after treatment (P=.03). Although statistical significance was not reached in the primary end point, the trial provides further support for IVIG as an efficacious adjunctive therapy in STSS. PMID- 12884157 TI - Dilemmas in the treatment of invasive Streptococcus pyogenes infections. PMID- 12884158 TI - Relationships between patient- and institution-specific variables and decreased antimicrobial susceptibility of Gram-negative pathogens. AB - The identification of patients infected with antibiotic-resistant strains of bacteria for inclusion in clinical trials remains a serious challenge for the future development of agents for use against such infections. To identify patient and institution-specific factors predictive of reduced susceptibility of Enterobacter species, Pseudomonas aeruginosa, and Klebsiella pneumoniae to cefepime, ciprofloxacin, and piperacillin-tazobactam, 5 years (1997-2001) of North American surveillance data were analyzed. The relationship between minimum inhibitory concentration (MIC) values for each organism-agent pair and patient- and institution-specific variables was analyzed using multivariable general linear modeling. The variables most commonly associated with decreases in susceptibility were duration of hospital stay before pathogen isolation, hospital size, primary diagnosis, and medical service. Combinations of these variables were associated with increases in observed MIC90 values of as much as 16-32-fold. Our findings demonstrate a relationship between MIC and certain patient- and institution-specific variables. Such data should be considered in the design of clinical trials directed at the study of resistant pathogens. PMID- 12884159 TI - Extensive limb swelling after immunization: reports to the Vaccine Adverse Event Reporting System. AB - Extensive limb swelling (ELS) has been reported after vaccination with a limited number of vaccine types. We sought to describe vaccine types involved in and the clinical characteristics of ELS cases reported to the Vaccine Adverse Event Reporting System (VAERS). A case of ELS was defined as any report of edema extending at least to the elbow or knee of a vaccinated extremity. Four hundred ninety-seven cases were identified, with some describing swelling from the shoulder to the hand or the hip to the foot. Patient age ranged from 0.1 to 91 years. The proportion of reports of ELS associated with a given vaccine, among all VAERS reports received for that vaccine, varied substantially among vaccines. Most reactions began within 1 day after vaccination and involved other signs of inflammation. Postvaccination ELS can involve both the proximal and distal segments of the extremity, affects all age groups, and occurs after vaccination with a broad range of vaccines. PMID- 12884160 TI - Listeriosis in patients at a comprehensive cancer center, 1955-1997. AB - Listeria monocytogenes infection occurred in 94 patients during 1955-1997 at Memorial Sloan-Kettering Cancer Center. The incidence was 0.5 (1955-1966), 0.96 (1970-1979), and 0.14 (1985-1997) cases per 1000 new admissions. Eighty-five patients (90%) were bacteremic, and 34 (36%) had evidence of intracranial infection. In 91 patients with cancer, 70 (77%) received chemotherapy for advanced or relapsed malignancy (n=51; 56%); 64 (68%) received corticosteroids. Breast cancer was the most common solid-organ cancer (n=14; 45%), and 34 (36%) had preexisting advanced liver disease. In 14 (39%) of 37 patients who died of listeriosis, death occurred within 48 h of L. monocytogenes isolation. Four (80%) of 5 patients with extracranial foci of infection died of their infection, compared with 33 (37%) of 89 patients with isolated bacteremia and/or intracranial infection (odds ratio, 2.34; P=.05). Most infections (60%) were due to L. monocytogenes serotype 1/2, and the remainder (40%) were due to serovar 4b. Listeriosis in these patients with cancer occurred most often in individuals receiving antineoplastic therapy for advanced or relapsed malignancy and systemic corticosteroids. The presence of advanced liver disease may have increased the risk of systemic listeriosis in susceptible patients with underlying cancer. PMID- 12884161 TI - Diagnostic accuracy of stool assays for inflammatory bacterial gastroenteritis in developed and resource-poor countries. AB - Because acute bacterial gastroenteritis is often inflammatory, rapid stool assays that detect intestinal inflammation might be used to distinguish between bacterial and nonbacterial gastroenteritis. We performed meta-analyses to determine the discriminatory power, in developed and in resource-poor countries, of rapid stool assays that test for lactoferrin, fecal leukocytes, fecal erythrocytes, and occult blood. In developed countries, the area under the summary receiver operating characteristic curve (AUC/SROC) was 0.89 for fecal leukocytes and 0.81 for occult blood. In resource-poor countries, the AUC/SROC was 0.79 for lactoferrin, 0.72 for fecal leukocytes, 0.63 for occult blood, and 0.61 for fecal erythrocytes. In developed countries, positive and negative likelihood ratios (LR+ and LR-, respectively) for fecal leukocytes were 4.56 and 0.32 when a threshold of >5 cells/high-power field was used, compared with 2.94 and 0.6 in resource-poor countries; for lactoferrin, LR+ was 1.34 and LR- was 0.17 in resource-poor countries when the threshold was an agglutination rating of "+" and a dilution of 1:50. In developing countries, rapid stool assays performed poorly, whereas in developed countries, tests for fecal leukocytes, lactoferrin, and occult blood were moderately useful and could identify patients who were more likely to benefit from empirical antibiotic therapy. PMID- 12884162 TI - Emergence of levofloxacin-resistant pneumococci in immunocompromised adults after therapy for community-acquired pneumonia. AB - We describe 4 patients infected with levofloxacin-resistant pneumococci after therapy for community-acquired pneumonia (CAP). The 4 patients had 15 episodes of CAP; Streptococcus pneumoniae was isolated from blood or sputum samples obtained during 14 of the episodes. The underlying medical condition was Bruton agammaglobulinemia in 3 patients and chronic lymphoid leukemia in the other. The initial episode of CAP in each patient was due to a levofloxacin-susceptible strain. One of 4 reinfections and 5 of 6 relapses were due to levofloxacin resistant strains. All of these strains had amino acid substitutions in the quinolone-resistance-determining region of the genes parC and gyrA. The time between episodes of pneumonia varied from 1 to 4 months. In immunocompromised patients with suspected or proven pneumococcal infection, it may be prudent not to use fluoroquinolone monotherapy empirically when the patient has a history of fluoroquinolone therapy in at least the past 4 months. PMID- 12884163 TI - Vancomycin versus placebo for treating persistent fever in patients with neutropenic cancer receiving piperacillin-tazobactam monotherapy. AB - This prospective, double-blind trial assessed whether the addition of a glycopeptide would be able to reduce the time to defervescence in neutropenic patients with cancer who had persistent fever 48-60 h after the initiation of empirical piperacillin-tazobactam monotherapy. Of 763 eligible patients, 165 with persistent fever were randomized to receive piperacillin-tazobactam therapy plus either vancomycin therapy or placebo. Defervescence was observed in 82 (95%) of 86 patients in the vancomycin group and in 73 (92%) of 79 patients in the placebo group (P=.52). The distributions of the time to defervescence were not statistically significant between the 2 groups (estimated hazard ratio, 1.03; 95% confidence interval, 0.75-1.43; P=.75). The number of additional episodes of gram positive bacteremia and the percentage of patients for whom amphotericin B was empirically added to their therapy regimen were also similar in both groups. This study failed to demonstrate that the empirical addition of vancomycin therapy to the treatment regimen is of benefit to persistently febrile neutropenic patients with cancer. PMID- 12884164 TI - Fish tank exposure and cutaneous infections due to Mycobacterium marinum: tuberculin skin testing, treatment, and prevention. AB - In the present study, 8 patients with soft tissue infection due to Mycobacterium marinum are described, and contemporary data on treatment are reviewed. Six patients had positive cultures, all patients had cutaneous exposure to fish tanks, 7 had sporotrichoid lesions, and 2 had deep infection. All 7 tested patients had tuberculin skin test reactions > or =10 mm. Six patients with disease limited to the skin were successfully treated with 2-drug combination therapy, including clarithromycin, ethambutol, and rifampin. Optimal treatment should include 2 drugs for 1-2 months after resolution of lesions, typically 3-4 months in total. Deeper infections may require more prolonged treatment and surgical debridement. Positive tuberculin reactions may be due to infection with M. marinum. Persons with open skin lesions or immunosuppression should avoid cutaneous contact with fish tanks. PMID- 12884165 TI - Cholera, diarrhea, and oral rehydration therapy: triumph and indictment. AB - Cholera drove the sanitary revolution in the industrialized world in the 19th century and now is driving the development of oral rehydration therapy (ORT) in the developing world. Despite the long history of cholera, only in the 1960s and 1970s was ORT fully developed. Scientists described this treatment after the discovery of the intact sodium-glucose intestinal cotransport in patients with cholera. This new understanding sparked clinical studies that revealed the ability of ORT to reduce the mortality associated with acute diarrheal disease. Despite the steady reductions in mortality due to acute dehydrating diarrheal diseases achieved by ORT, the costly morbidity due to these diseases remains, the result of a failure to globalize sanitation and to control the developmental impact of diarrheal diseases and their associated malnutrition. New advances in oral rehydration and nutrition therapy and new methods to recognize its costs are discussed in this review. PMID- 12884166 TI - Human illness associated with use of veterinary vaccines. AB - Veterinary vaccines are being used with increasing frequency in the United States to protect the health of animals. However, humans may be inadvertently exposed to these products by means of unintentional inoculation or other routes of exposure. The potential for both exposure and for adverse consequences secondary to exposure to veterinary vaccines may be growing. With the exception of brucellosis vaccines, there have been few reports of suspected or confirmed adverse events in humans associated with the use of animal vaccines, but it is unclear whether that is because few adverse events occur or because adverse events are not recognized and/or reported. Results of a search for relevant literature and of communications with health officials at governmental and private institutions suggest that enhanced efforts are needed to recognize and to prevent human illness associated with use of veterinary vaccines. PMID- 12884167 TI - Amphotericin B: time for a new "gold standard". AB - When introduced in 1959, amphotericin B deoxycholate (AmBD) was clearly a life saving drug. Randomized studies demonstrating its efficacy were not thought to be necessary, and it was granted indications for many invasive fungal infections. Despite its formidable toxicities, AmBD is thus often used as the primary comparator in studies of invasive fungal infections. Safer lipid-based versions of amphotericin B (AmB) have been introduced, but difficulties with studying these agents generally led to licensure for salvage therapy, not primary therapy. However, the cumulative clinical experience to date with the lipid-based preparations is now adequate to demonstrate that these agents are no less active than AmBD, and, for some infections, it can now be stated that specific lipid based preparations of AmB are superior to AmBD. Given their superior safety profiles, these preparations can now be considered suitable replacements for AmBD for primary therapy for many invasive fungal infections in clinical practice and research. PMID- 12884168 TI - Separate worlds set to collide: smallpox, vaccinia virus vaccination, and human immunodeficiency virus and acquired immunodeficiency syndrome. AB - Concerns about the possible release of smallpox by bioterrorists has led to policies that recommend smallpox vaccination of some health care providers, and, in the near future, the vaccine may become available to the general population on a voluntary basis. Both smallpox virus (variola virus) and the smallpox vaccine (vaccinia virus) will have a significant impact on people infected with human immunodeficiency virus (HIV). Given that populations with acquired immunodeficiency syndrome and populations with immunosuppressed conditions due to solid organ and bone marrow transplantation were not present in the days when smallpox was prevalent, we will speculate on how smallpox might present in immunodeficient patients, and we will review the adverse events expected from the smallpox vaccine in hosts with HIV infection. PMID- 12884169 TI - Do sex and race/ethnicity influence CD4 cell response in patients who achieve virologic suppression during antiretroviral therapy? AB - To determine whether CD4 cell count response to virus suppression during highly active antiretroviral therapy differs according to sex or race/ethnicity, we analyzed data in our observational cohort study for patients receiving their first antiretroviral regimen who experienced virus suppression by 6 months of therapy. In both univariate and multivariate analyses, women had greater CD4 cell count increases, compared with men, as did patients receiving a regimen that did not include a protease inhibitor. Race/ethnicity was not a factor. PMID- 12884170 TI - Response of human immunodeficiency virus-infected patients receiving highly active antiretroviral therapy to vaccination with 23-valent pneumococcal polysaccharide vaccine. AB - Whether highly active antiretroviral therapy (HAART) impacts responses to 23 valent pneumococcal polysaccharide vaccine (PV) is not known. Immunoglobulin G (IgG) levels for 6 capsular polysaccharides in human immunodeficiency virus (HIV) infected patients who had received > or =6 months of HAART were measured either after their first dose of PV (n=46) or after revaccination (n=41); control subjects had never received HAART and had received the first dose of PV (n=38). There were no significant differences in pre- or postvaccination IgG levels among these groups but for 1 capsular polysaccharide. The 3 groups had significant postvaccination increases in IgG levels to all capsular polysaccharides. The control group had a greater number of 2-fold responses than did the combined HAART groups (P<.05). Patients with a CD4 cell count of > or =200 cells/mm3 had a greater number of 2-fold responses than did those with a CD4 cell count of <200 cells/mm3 (P<.05). For revaccinated patients, postvaccination IgG levels were correlated with the CD4 cell count at the initial vaccination. The immunogenicity of PV among patients receiving long-term HAART is modest. It seems best to immunize HIV-infected patients early in the course of disease. PMID- 12884171 TI - Genetic confirmation of atovaquone-proguanil-resistant Plasmodium falciparum malaria acquired by a nonimmune traveler to East Africa. AB - We report a case of atovaquone-proguanil-resistant Plasmodium falciparum malaria acquired by a nonimmune traveler to Kenya. Recurrent parasitemia occurred 30 days after directly observed therapy with a combination of atovaquone and proguanil. Treatment failure was confirmed by genetic fingerprinting and sequencing. The primary isolate had wild-type sequence of cytochrome b; however, the recrudescent isolate had a single mutation at position 268 (Tyr268Ser). PMID- 12884172 TI - Five years' experience of reverse-transcriptase polymerase chain reaction in daily diagnosis of enterovirus and rhinovirus infections. AB - To determine the efficiency of reverse-transcriptase polymerase chain reaction (RT-PCR) assays currently used in diagnosing enterovirus and rhinovirus infections, we compared results obtained with RT-PCR methods, which detect both enteroviruses and rhinoviruses simultaneously, with results obtained by conventional virus isolation. Both tests were performed on 591 specimens: 38 samples (6%) had positive results by both RT-PCR and isolation, 90 samples (15%) had positive results by RT-PCR only, and 7 samples (1%) had positive results only by virus isolation. In conclusion, RT-PCR was superior in rapidity and sensitivity to virus isolation and is recommended as the primary diagnostic tool for enterovirus and rhinovirus infections. PMID- 12884173 TI - A possible case of myelopathy/tropical spastic paraparesis in an Argentinian woman with human T lymphocyte virus type II. AB - A case of neurological disease featuring human T lymphocyte virus-associated myelopathy/tropical spastic paraparesis (HAM/TSP) was diagnosed by serological (Western blot) and molecular (polymerase chain reaction) criteria as related to human T lymphocyte virus (HTLV)-II infection. This is, to our knowledge, the first case of this kind found in Argentina and is additional evidence that HAM/TSP solely related to HTLV-II infection occurs in HTLV-I-negative subjects. PMID- 12884174 TI - Mortality rates associated with methicillin-resistant and -susceptible Staphylococcus aureus infections. PMID- 12884176 TI - Impact of hepatitis C virus (HCV) on morbidity and mortality rates among HIV infected patients. PMID- 12884178 TI - Evaluation of current immunodiagnostic criteria for diagnosis of neurocysticercosis. PMID- 12884179 TI - Pruritic skin eruption on the left foot of a 36-year-old woman. PMID- 12884180 TI - Should treatment of hepatitis C in HIV-seropositive and HIV-seronegative patients with hemophilia include induction doses of interferon? PMID- 12884182 TI - Antimicrobial resistance of Salmonella enterica serotype Typhi in Dakar, Senegal. PMID- 12884183 TI - Community-acquired methicillin-resistant Staphylococcus aureus. PMID- 12884184 TI - Smallpox vaccination after a bioterrorism-based exposure. PMID- 12884185 TI - Failure of linezolid treatment for enterococcal endocarditis. AB - We describe a patient with Enterococcus faecalis endocarditis who failed to respond to treatment with linezolid. PMID- 12884186 TI - Infective endophthalmitis caused by Bacillus cereus after cataract extraction surgery. AB - We describe a 72-year-old Chinese man who developed fulminant Bacillus cereus endophthalmitis 36 h after an uneventful surgery for cataract extraction. Clinical progression of disease was extremely rapid, in spite of early vitrectomy and intravitreal injection of antibiotics. Results of Gram staining showed gram positive bacilli, and the culture was positive for B. cereus. Endophthalmitis is rare after cataract surgery (cumulative probability within 1 year, approximately 0.08%), and cases caused by B. cereus are even less common. The prognosis and plan of treatment are discussed. PMID- 12884187 TI - Isolated pulmonary Mycobacterium avium complex infection in patients with human immunodeficiency virus infection: case reports and literature review. AB - We report 4 cases of isolated pulmonary Mycobacterium avium complex (MAC) infection and review the 20 previously reported cases in the human immunodeficiency virus literature. All 4 patients had acquired immune deficiency syndrome, and 3 were believed to have had an immune reconstitution syndrome as a cause of MAC infection. Two patients underwent bronchoscopy with biopsy, revealing endobronchial lesions and granuloma formation, and all 4 patients responded well to MAC therapy. PMID- 12884188 TI - Acute renal failure associated with tenofovir treatment in a patient with acquired immunodeficiency syndrome. AB - We report a case of acute renal failure due to proximal tubular necrosis associated with tenofovir treatment in a patient with acquired immunodeficiency syndrome. PMID- 12884189 TI - Liver transplantation in neonates. AB - Orthotopic liver transplantation (OLT) has evolved over the past two decades to become the standard of care for end-stage liver disease in infants and children. Technical advances, particularly the use of technical variant allografts, have permitted extension of OLT into a much younger and smaller population than previously possible. Major centers around the world now routinely perform OLT in infants with survival success equivalent to that in older children and adults. We are beginning to see a small population of school-aged children who were infant OLT recipients. The further extension of OLT into neonates is more recent, with only a few pediatric centers reporting survival success. Very little is known about this frontier of transplantation. Our intent is to provide an overview of neonatal OLT using all available data and our experience in the field. PMID- 12884190 TI - Arterialized partial orthotopic liver transplantation in the mouse: a new model and evaluation of the critical liver mass. AB - The availability of a model of partial orthotopic liver transplantation (OLT) in the mouse would be an important tool for studying injuries associated with transplantation. The goals of this study were three-fold: (1). to develop a model of partial OLT in the mouse, (2). to determine the minimal graft volume in this model, and (3). to define the injury associated with small volume incompatible with animal survival. Putative grafts of 30% and 50% were prepared. Their weight was 30 +/- 5% and 45 +/- 10%, respectively. Subsequently, 30% and 45% syngeneic partial liver grafts were orthotopically transplanted into C57BL/6 mice. Each recipient receiving a 45% graft survived permanently, whereas those receiving only a 30% graft volume died within 2 to 4 days of surgery. Serum transaminase levels normalized in the 45% graft group within 14 days after surgery. In this group, small foci of necrosis and mild steatosis were noted on histology at postoperative day 2, but no abnormalities were noted after 14 days and 100 days. In contrast, recipients who underwent transplantation with a 30% graft volume showed a comparable amount of necrosis and significant microvesicular steatosis in most hepatocytes 2 days after surgery. Hepatocyte proliferation was reduced in this group when compared with animals who underwent transplantation with a 45% graft volume. In conclusion, partial liver transplantation is feasible in the mouse with a critical graft volume ranging between 30% and 45%. Small liver grafts develop massive microvesicular steatosis and impaired regeneration rapidly leading to animal death. PMID- 12884191 TI - Outcome of liver transplantation for patients infected by hepatitis C, including those infected by genotype 4. AB - Predictors of hepatitis C virus (HCV)-related liver disease posttransplantation are still unclear. The impact of HCV genotype on outcome of transplantation has been studied, but conclusions are not in agreement. The role of HCV genotype 4 on the result of liver transplantation requires further study. The aim of this study is to examine the outcome of liver transplantation for patients with HCV genotype 4 infection. The study group included 128 patients who underwent transplantation for HCV infection: 28 patients, genotype 1; 11 patients, genotype 2; 19 patients, genotype 3; and 32 patients, genotype 4. For 64 of 128 patients, genotype was known and an assessable histological specimen was available. Median interval from transplantation to biopsy was 1.92 years (range, 0.24 to 11.48 years). Twenty-six percent of HCV genotype-4 patients developed either severe fibrosis or cirrhosis versus 6.7% in the genotype non-4 group (P =.04). A statistically significant greater fibrosis progression rate was observed in genotype-4 than genotype non-4 patients. In univariate and multivariate analysis, rapid liver fibrosis was associated with the presence of HCV genotype-4 infection. In addition, donor and recipient age and graft warm ischemic time also were associated with rate of fibrosis progression. Five-year cumulative rates for the development of cirrhosis or severe liver fibrosis were 84% in genotype-4 and 24% in genotype non-4 patients (P =.02). Five-year survival rates for patients with genotypes 1, 2/3, and 4 were 72%, 80%, and 79%, respectively (P =.8). In conclusion, 5-year survival for patients who underwent transplantation for HCV genotype-4 infection was similar to that of genotype non-4 patients; however, more severe fibrosis and rapid fibrosis progression was observed after transplantation in patients with genotype-4 infection. PMID- 12884192 TI - Prolonged survival of rat liver allograft with adenoviral gene transfection of human immunodeficiency virus type 1 nef. AB - HIV-1 nef is believed to allow immune evasion by modifying cell surface molecules because of certain mechanisms such as downregulation of the major histocompatibility complex (MHC) class I molecule complex as well as upregulation of FasL. In the present study, we successfully generated a recombinant adenovirus vector containing HIV-1 nef. We detected the expression of nef in liver infected with AxCANef by immune staining and Western blotting, and confirmed its expression as persistent for more than 4 weeks. Furthermore, the surface expression of MHC class I was downregulated in AxCANef-infected hepatic cells. In addition, we also observed nef-induced FasL upregulation of gene-transfected hepatic cells. Using a DA-to-Lewis orthotopic liver transplantation model, we transfected AxCANef to a liver graft to determine whether nef expression could have an effect on recipient survival. AxCANef significantly prolonged recipient survival time (14.5 days) compared with the uninfected group (11 days) (P <.001) and the AxCALacZ-infected group (11 days) (P <.001). Histologic analysis showed reduction in the number of accumulated inflammatory cells and an increase in apoptotic cells in grafts expressing nef. In conclusion, we showed that the nef gene could prolong survival of rat liver allografts, and this result suggested the potential clinical use of its transfection. PMID- 12884193 TI - A multicenter pilot prospective study comparing Celsior and University of Wisconsin preserving solutions for use in liver transplantation. AB - Primary dysfunction (PDF) still occurs after orthotopic liver transplantation (OLT). Celsior solution (CS) might offer some advantages over the conventional University of Wisconsin (UW) solution for organ preservation, but to date, this has not been prospectively evaluated in the context of OLT. In this prospective, randomized, multicenter, pilot study, 215 potential liver donors were enrolled and randomized. In 42 cases, the livers were unsuitable for transplantation; therefore, 173 randomized livers ultimately were implanted after perfusion and cold preservation with CS (n = 83) or UW solution (n = 90). In accord with the indications of the CS manufacturing company, total CS infusion volume was 90 mL/kg, greater than that of UW solution (60 mL/kg). The main aim of the study is to compare the prevalence of PDF between the two groups. Donor and recipient variables were similar in the two groups. Episodes of PDF were numerically lower in the CS (2.4%) than UW group (7.8%), but the difference was not statistically significant. There was a trend toward a lesser need for early re-OLT (<30 days) in the CS group (P =.0507), but again, no statistically significant difference emerged. Overall and time-differentiated postoperative deaths also were similar. One-year actuarial patient (UW, 89% v CS, 87%) and graft (UW, 83% v CS, 85%) survival rates were similar. In conclusion, CS was similar to UW solution as a preservation solution in the clinical setting of OLT at the infusion volumes described, although some theoretical advantages of CS composition suggest that CS might prove a valid alternative to UW preservation solution in multiorgan harvesting, including the liver. A study on a larger patient basis is needed. PMID- 12884194 TI - Histidine-tryptophan-ketoglutarate versus University of Wisconsin solution in living donor liver transplantation: results of a prospective study. AB - The grafts obtained from a living donor hepatectomy are perfused on the back table with either University of Wisconsin solution (UW) or histidine-tryptophan ketoglutarate solution (HTK). The efficacy and safety of these solutions have been studied in cadaveric liver transplantation, however, there is no study comparing the two solutions in adult-to-adult living donor liver transplantation. In this study, UW and HTK were used in the perfusion of right living donor grafts. The grafts were perfused with a predetermined sequence and volume of one of the solutions. Liver biochemistries, complications, and graft and patient survival were analyzed. From January 2001 to September 2002, 30 grafts were alternately perfused with either UW (UW group) or HTK (HTK group). The perfusion was performed first via the artery and then via the portal vein with a predetermined volume. At a mean follow-up of 13 +/- 7 months, no significant statistical difference between groups UW and HTK in posttransplantation liver biochemistries, complications, or patient and graft survival (84% and 80%, respectively) was observed. In conclusion, UW and HTK are equally effective and safe in the perfusion of the living donor liver grafts. HTK has a slight practical advantage over UW because it does not need to be flushed away before reperfusion of the graft and is less expensive. PMID- 12884195 TI - Safety and outcome of hepatitis B core antibody-positive donors in right-lobe living donor liver transplantation. AB - We studied the safety of right-lobe donor hepatectomy in living donors with a positive serological test result for antibody against hepatitis B core antigen (anti-HBc). Of 54 right-lobe living liver donors, serum anti-HBc was positive in 29 donors (54%) and negative in 25 donors (46%). None had a history of hepatitis and all had normal liver biochemistry test results before surgery. Surgical data, postoperative liver function, and outcome of donors were compared according to anti-HBc status. Donors positive for anti-HBc were significantly older (median age, 42 v 31 years; P <.001), but there was no difference in other demographics and liver size. Median blood loss was greater in anti-HBc-positive donors (600 v 350 mL; P =.007). Histological examination showed no difference in degree of portal fibrosis or fatty change in liver grafts. There was no significant difference in postoperative serum transaminase levels or prothrombin times; however, anti-HBc-positive donors had greater serum bilirubin levels day 6 (26 v 21 micromol/L; P =.01) and day 7 (22 v 15 micromol/L; P =.004) after surgery. Postoperative complications developed in 10 anti-HBc-positive and 6 anti-HBc negative donors (P =.4). All donors who developed cholestasis were positive for anti-HBc, aged 45 years or older, and had mild fatty changes of the liver. Hospital stays were similar. All donors had completely normal liver function at a median follow-up of 31 months (range, 21 to 76 months). A positive serological test result for anti-HBc should not be regarded as a contraindication for right lobe liver donation. PMID- 12884196 TI - Are we ready for marginal hepatitis B core antibody-positive living liver donors? PMID- 12884197 TI - Use of hepatitis B core antibody-positive donors in recipients without evidence of hepatitis B infection: a survey of current practice in the United States. AB - Because of the current organ shortage, some liver transplant programs have begun to accept marginal organs that previously would have been rejected. An example is the use of donors with evidence of past hepatitis B virus (HBV) infection. To gain insight into the use of hepatitis B core antibody-positive (anti-HBc(+)) donor livers in recipients without evidence of HBV infection, we conducted a survey. Surveys consisting of 12 multiple-choice questions were sent to all 110 liver transplant programs across the United States in mid-2001, and 56 of 110 surveys (51%) could be evaluated. Overall, 32 of 56 programs (57%) indicated they would transplant an anti-HBc(+) liver into a recipient without serological evidence of HBV infection. Of those who would accept an anti-HBc(+) liver, 16 of 27 respondents (59%) indicated knowledge of HBV DNA status would change their protocol; 46% of these respondents would decrease prophylaxis if HBV DNA was negative, 27% would increase prophylaxis if HBV DNA was positive, and 27% would not accept the liver if HBV DNA was positive. Conversely, 9 of 28 respondents (32%) who would not accept an anti-HBc(+) liver stated that knowing HBV DNA status would change their protocol in that they might consider accepting livers if HBV DNA was negative. In conclusion, as of mid-2001, of transplant medical directors in the United States who responded to our survey, 57% would accept an anti-HBc(+) donor liver for an HBV-naive recipient. Treatment protocols for using these organs varied. Knowledge about HBV DNA status of the donor and/or liver would greatly influence prophylaxis for those accepting anti-HBc(+) donor livers. PMID- 12884198 TI - Incidence and prevalence of coccidioidomycosis in patients with end-stage liver disease. AB - Coccidioidomycosis is an endemic fungal infection of the desert southwestern United States. Patients with end-stage liver disease (ESLD) have not been described as having a high rate of coccidioidal infection. We prospectively evaluated 290 patients with ESLD for liver transplantation and found that 6 of these patients (2.1%) had active coccidioidal infection at presentation. Of 184 patients listed for transplantation, 48 patients were observed for at least 1 year; among these 48 patients, new coccidioidal infection developed in 2 patients within the year, for a 1-year incidence of 4.2%. Conversely, the incidence of coccidioidal infection in Maricopa County, Arizona in that same period was 0.04%. Awareness is needed to identify the presence of coccidioidal infection, which may be masked by symptoms and findings of ESLD. Treatment may alleviate some of the symptoms of coccidioidomycosis originally attributed to the concomitant liver disease. PMID- 12884199 TI - Dual-echo, chemical shift gradient-echo magnetic resonance imaging to quantify hepatic steatosis: Implications for living liver donation. AB - In living liver donation, a fatty liver poses risks for both recipient and donor. Currently, liver biopsy is the standard for assessing the presence and extent of steatosis. The goals of this study were to correlate a steatosis index derived from magnetic resonance imaging (MRI) to the histologic grade on biopsy as well as to determine the topographic distribution of steatosis within the liver. We examined the ability of dual-echo, chemical shift gradient-echo MRI to predict the degree of steatosis on liver biopsy. A total of 22 subjects received both a liver biopsy and detailed MRI evaluation. These individuals included 15 potential living donors and 7 patients with nonalcoholic fatty liver disease. MRI steatosis index was then compared with histologic grade on liver biopsy. The topographic distribution of hepatic steatosis was determined from those subjects in whom MRI detected hepatic steatosis. The steatosis index had a positive correlation with grade of steatosis on liver biopsy (correlation coefficient, 0.84). There was no significant variation in the degree of steatosis among segments. A steatosis index of >0.2 had good positive and negative predictive value for the presence of significant steatosis (>15%) on biopsy. Our quantitative MRI protocol can predict the degree of hepatic steatosis when it is minimal to moderate, and may obviate the need for liver biopsy for the purpose of quantification of steatosis in living donors. Fat saturation added to the MRI protocol may further improve diagnostic accuracy. This technique may be applicable to the larger population with hepatic steatosis. PMID- 12884200 TI - Serial measurements of bone density at the lumbar spine do not predict fracture risk after liver transplantation. AB - Bone disease has emerged as a serious and complex complication after liver transplantation. The purpose of this study is to determine risk factors for fracture and bone loss after liver transplantation. Dual-energy x-ray absorptiometry (DEXA) of the lumbar spine was performed routinely pretransplantation, 6 months posttransplantation, and at yearly intervals thereafter at our center. We followed up patients who underwent transplantation in the past 10 years and compared bone mineral density (BMD) and fracture rate with known risk factors for bone loss in primary transplant recipients who met the inclusion criteria of a pretransplantation DEXA and at least one follow-up DEXA scan postoperatively (n = 153). We observed a 15% (n = 23) prevalence of symptomatic fractures at a mean of 2.2 +/- 1.8 years after transplantation. Change in BMD was greatest from pretransplantation to 6 months posttransplantation (-4.2%; P =.006), then increased at a rate of 1.4% per year. Logistic regression analysis showed an association of fracture risk with several factors, including number of acute rejection episodes (P =.045), smoking (P =.02), and female sex (P =.02). Stepwise logistic regression analysis reported female sex (P =.004) as the only factor associated with fracture after transplantation. Age, time listed for transplantation, race, menopause, chronic renal insufficiency, loss of height, family history of osteoporosis, BMD, and T score did not predict fracture or bone loss after transplantation. In conclusion, serial measurements of BMD at the lumbar spine do not appear to predict fracture risk; however, data suggest that female sex is the strongest predictor of fracture after liver transplantation. PMID- 12884201 TI - Impact of right-lobe live donor liver transplantation on patients waiting for liver transplantation. AB - Right-lobe live donor liver transplantation (LDLT) is accepted for adult patients, but the incremental benefit of LDLT over cadaveric donor liver transplantation (CDLT) is unknown. We evaluated prospectively the impact of right lobe LDLT on patients listed for CDLT. Between January 1999 and December 2001, a total of 152 patients with chronic liver disease were listed for liver transplantation. The options of LDLT and CDLT were given after listing. Sixty five patients (43%) had volunteers and 87 patients (57%) did not have volunteers. The groups with and without volunteers were similar in terms of age, diagnosis of liver disease, and Model for End-Stage Liver Disease score. The group with volunteers had a shorter waiting time for transplantation (median, 38 v 344 days; P <.001), greater transplantation rate (72% v 20%; P <.001), and lower mortality rate while waiting for a transplant (15% v 30%; P =.037). Overall, right-lobe LDLT was performed in 44 patients (29%). It increased the transplantation rate from 13% to 42%. On follow-up until December 2002 (median follow-up, 17.4 months), survival rates of the groups with and without volunteers were 68% and 51%, respectively (P =.034). One- and 3-year survival rates were 72.3% and 66.4% for the group with volunteers and 61.9% and 45.6% for the group without volunteers, respectively (P =.028). In conclusion, right-lobe LDLT offers patients listed for transplantation an incremental benefit by tripling the transplantation rate and improving overall survival. PMID- 12884202 TI - Recurrent deep-vein thrombosis based on homozygous factor V Leiden mutation acquired after liver transplantation. AB - Several genetic liver diseases can be treated by liver transplantation (LT). However, some genetic defects also may be acquired by this procedure. We describe a patient who developed recurrent deep-vein thromboses after LT for hepatitis C virus-associated hepatocellular carcinoma on the basis of a homozygous Leiden mutation of the factor V gene in the donor liver. Liver donors with a history of venous thrombosis should be screened for the presence of activated protein C (APC) resistance. In addition, we recommend looking for APC resistance in liver recipients who develop venous thromboembolic disease in the post-LT course. Molecular analysis of donor tissue may be necessary to make a definite diagnosis of factor V Leiden mutation in these patients. As a consequence, intensified postoperative thromboprophylaxis or lifelong anticoagulant therapy may be necessary if this thrombophilic gene defect is detected. PMID- 12884203 TI - Cavoportal transposition for portal vein thrombosis in a pediatric living-related liver transplantation. AB - We report a case of a living-related liver transplantation in a child in whom diffuse thrombosis of the portal venous system required the use of recipient vena cava to perfuse the donor portal vein (cavoportal transposition). An 8 (1/2) month-old infant with biliary atresia received the left lateral segment of his father's liver. The child's portal vein was irretrievably thrombosed, as were the splenic and superior mesenteric veins. A cavoportal anastomosis provided excellent flow to the liver, and there was no engorgement of the graft, which had good immediate function. The main postoperative problem was loss of 3 to 3.5 liters of ascitic fluid through the drain for 2 weeks. The ascites eventually resolved over a period of 4 weeks. Twenty-four months after transplantation, the child is thriving on a normal diet and has no ascites. A Doppler examination showed good flow in the cavoportal anastomosis. PMID- 12884204 TI - Kaposi's sarcoma in a liver graft recipient. PMID- 12884205 TI - Setting organ allocation priorities: should we care what the public cares about? AB - OBJECTIVE: To investigate the nature of public preferences in the allocation of donor liver grafts for transplantation. DESIGN: A qualitative study based upon the transcripts of four focus groups. SETTING: Derby, Derbyshire, UK. PARTICIPANTS: Twenty-two members of the public in the Derby locality, recruited to one of four focus groups through local community groups. MAIN OUTCOME MEASURES: The views of focus group members as to the importance (or otherwise) of several potential discriminating factors which could be used in the prioritization of patients on the waiting list for liver transplantation were ascertained. The factors included were expected posttransplantation prognosis, the age of the patient, whether the patient was personally responsible for their illness, the time spent on the waiting list, re-transplantation or primary transplant and the social background of the patient. RESULTS AND CONCLUSIONS: Group members explored the criteria from a number of perspectives, and made some unexpected linkages between the criteria and wider moral principles. They did not come to firm conclusions about the relative desirability of the criteria, but their approach was notably flexible and thoughtful, with the exception of a few instances where they appeared to resort to arguments based on what is 'obvious' and 'natural'. The results of these discussions suggest that members of the public would be able and willing to respond positively to a more open and consultative system of donor liver prioritization than exists presently within the UK. PMID- 12884206 TI - Multivisceral and isolated intestinal procurement techniques. AB - Improved outcomes in multivisceral and isolated intestinal transplantation have generated increased demand for these procedures. Enhanced recognition of potential multivisceral/intestinal donors and widespread application of advanced organ procurement techniques is necessary in the current climate of organ scarcity. This manuscript details the multivisceral and isolated intestinal procurement techniques currently performed at the University of California Los Angeles. PMID- 12884207 TI - Serum protein S-100b in acute liver failure: Results of the US Acute Liver Failure Study group. PMID- 12884208 TI - Has cerumen a protective role in recurrent external otitis? AB - PURPOSE: We investigated the bactericidal activity of the cerumen in patients with recurrent otitis externa. MATERIALS AND METHODS: Cerumen samples were collected from 2 groups. Group A (n = 20) consisted of patients with recurrent otitis externa (2 or more acute otitis externa attack in the current year) and group B (n = 30) consisted of cerumen from a healthy population. We examined the bactericidal activity against the common microorganisms (Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, Pseudomonas aeruginosa, and Enterococcus) that encounter the recurrent otitis externa. RESULTS: A significant decrease in the count of S epidermidis was observed in group A (P <.05) and B (P <.01) was observed. A comparison of decreases and increases in the percentages of microorganisms between the groups A and B showed that there was a significant difference only in the count of E coli (P 12 pg/ml) were correlated with larger tumor size, elevated serum CRP levels, and liver metastases (P < 0.05). IL-6 levels also increased in a stage-related manner (P < 0.01). Although serum IL-6 correlated with survival, it is not an independent prognostic indicator. CONCLUSIONS: Serum IL-6 levels correlated with disease status of colorectal cancer but could not be regarded as an independent predictor for prognosis. PMID- 12884235 TI - Color multiplex polymerase chain reaction for quantitative analysis of epidermal growth factor receptor genes in colorectal adenocarcinoma. AB - BACKGROUND AND OBJECTIVES: Epidermal growth factor receptor (EGFR) is a 170-kDa transmembrane cell surface receptor, which belongs to the c-erbB family of tyrosine kinases. Chimeric anti-EGFR monoclonal antibody is an investigational therapy for advanced adenocarcinoma of the colon. Anti-EGFR is believed to be most effective against those neoplasms with elevated EGFR levels. Possible mechanisms for over expression of EGFR include gene amplification, transcriptional upregulation, or decreased degredation of the EGFR protein. METHODS: We analyzed a series of 36 specimens of colonic adenocarcinoma with known levels of EGFR protein expression for amplification of the gene sequence of EGFR. Carcinomas were considered positive for EGFR expression when greater than 10% of the neoplastic cells stained at a level of 2+ or 3+. Foci of adenocarcinoma were microdissected from paraffin sections and quantitative real time PCR (polymerase chain reaction) performed using a thermal cycler with real time fluorescence capability (Light cycler(TM), Roche Diagnostics, Indianapolis, IN). A relative quantitation assay comparing the EGFR gene to the control albumin gene was performed by 2-color multiplexing. RESULTS: Usable data on gene amplification status were obtained in 31 of the 36 samples. The average EGFR/albumin gene copy number ratio for the 31 samples of colon adenocarcinoma in which PCR results were obtained was 1.13 +/- 0.55 with a range of 0.26-2.29. The average EGFR/albumin gene copy number ratio obtained for 16 normal DNA leukocyte samples used to establish the efficiency curves was 1.03 +/- 0.31 with a range of 0.49-1.51. CONCLUSIONS: EGFR protein is overexpressed in a significant percentage of colonic adenocarcinomas. As with Her-2/neu overexpression, overexpression of EGFR serves as a basis for specific antibody therapy in a subset of carcinomas. Unlike Her-2/neu, where most overexpression is secondary to gene amplification, overexpression of EGFR appears to be unrelated to gene amplification. PMID- 12884236 TI - BRCA1 expression in leukoplakia and carcinoma of the tongue. AB - BACKGROUND AND OBJECTIVES: Expression of BRCA1 was examined in patients with leukoplakia and carcinoma of the tongue. Its prognostic value was evaluated in patients with tongue cancer. METHODS: Expression of BRCA1 was studied by immunohistochemical localization. Cytoplasmic staining of BRCA1 was observed in both leukoplakia and carcinoma of the tongue. RESULTS: In leukoplakia, 61% and 39% of the patients expressed BRCA1 expression with a staining intensity of 1+ and 2+, respectively. In patients with hyperplasia (67%), BRCA1 expression with a staining intensity of 1+ was 67%; BRCA1 expression with a staining intensity of 2+ was 33%. In patients with dysplasia (33%; mild and moderate), BRCA1 expression with a staining intensity of 1+ and 2+ was 50% each. In carcinoma of the tongue, only 34% of the patients showed BRCA1 expression. In this group, 33% of the tumors exhibited 1+ staining, and only 1% of the tumors expressed 2+ staining. Moreover, BRCA1 expression with a staining intensity of 2+ was significantly higher in patients with dysplasia (50%) than in those with hyperplasia (33%), followed by patients with squamous cell carcinoma of the tongue (1%). The percentage positivity of BRCA1 expression in tongue cancer patients was significantly lower (34%), as compared with patients with leukoplakia (100%; P = 0.000001). A significant positive correlation was noted between BRCA1 and c-myc (P = 0.012). Univariate survival analysis by log-rank test and multivariate survival analysis by Cox regression showed that BRCA1 expression was the most significant prognostic factor predicting relapse-free survival of early-stage patients. CONCLUSIONS: Subcellular localization of the BRCA1 gene product provided evidence of its involvement in the pathogenesis of tongue tumors. PMID- 12884237 TI - Prognostic implication of cyclin E expression and its relationship with cyclin D1 and p27Kip1 expression on tissue microarrays of node negative breast cancer. AB - BACKGROUND AND OBJECTIVES: Altered expression of cell-cycle regulators is prevalent in clinical breast cancer. This study was performed to analyze the impact of cyclin E expression to the outcome of breast cancer together with cyclin D1 and p27Kip1. METHODS: The correlation between cyclin D1/E and p27Kip1 expression was analyzed in tissue arrays of 175 node-negative breast cancers treated by the same chemotherapy composed of fluorouracil, cyclophosphamide, and methotrexate. Data from the immunohistochemical assays of three molecules were correlated and were analyzed with clinical outcome of the patients. RESULTS: Cyclin E expression was observed in 48 (27.4%) of 175 breast carcinomas. Cyclin E expression was significantly increased in young age patients and poorly differentiate tumors. Expression of cyclin E was significantly increased in cyclin D1 expressing tumors (P = 0.034). p27Kip1 expression was preserved above the 50% level in 87 tumors (49.7%) and was inversely correlated with cyclin E expression (P = 0.042). Ki67 labeling index was significantly increased in cyclin E-expressing tumors (P = 0.033) and was inversely related with p27Kip1 expression. In multivariate survival analysis, cyclin E expression was significant for the prediction of poor survival of the patients. CONCLUSIONS: Cyclin E expression was associated with poor prognosis and intimately correlated with the expression of cyclin D1 and p27Kip1. Integration of TMA technology allowed a high-throughput analysis for correlating molecular in situ findings with clinico-pathologic information. PMID- 12884238 TI - Elevation of preoperative serum C-reactive protein level is related to poor prognosis in esophageal squamous cell carcinoma. AB - BACKGROUND AND OBJECTIVES: An increased serum C-reactive protein (CRP) level was found in patients with various malignant tumors and was associated with poor prognosis. The aim of this study was to analyze the clinicopathological significance and the prognostic value of preoperative CRP levels in patients with esophageal squamous cell carcinomas. PATIENTS AND METHODS: The preoperative CRP level was measured by enzyme-linked immunosorbent assay (ELISA) in 150 patients with primary esophageal squamous cell carcinomas. All patients underwent radical surgery without any preoperative therapy. The patients were divided into two groups using a cut-off value of 1.0 mg/dl. The pathological classifications of the tumor were examined according to the TNM/UICC classification. The associations between the clinicopathological factors and CRP level were determined. The prognostic value of CRP was determined using Cox's proportional hazards model. RESULTS: Thirty-five patients (23%) showed high CRP levels (more than 1.0 mg/dl). Statistically significant differences in CRP levels were observed depending on tumor depth (P = 0.022) and TNM/UICC stage (P = 0.001). A high CRP level was associated with poor survival (P = 0.005) and was confirmed by multivariate analysis. CONCLUSIONS: A high CRP level is associated with tumor progression and poor survival in patients with esophageal squamous cell carcinoma. PMID- 12884239 TI - Role of MUC1 and MUC5AC expressions as prognostic indicators in gastric carcinomas. AB - BACKGROUND AND OBJECTIVES: The aim of this study is to clarify the relationship between the expression of MUC1 and MUC5AC mucins and the clinicopathological features in human gastric carcinomas using the mouse monoclonal antibodies VU-4H5 and Clone 45M1, respectively. Furthermore, the possibility of using phenotypes (MUC1+/MUC5AC+, MUC1+/MUC5AC-, MUC1-/MUC5AC-, MUC1-/MUC5AC+) to predict prognosis of the patients is evaluated. METHODS: Formalin-fixed, paraffin wax-embedded tissues from 76 cases of gastric cancer were examined for the expression of MUC1 and MUC5AC mucin antigens immunohistochemically using the avidin-biotin peroxidase method. RESULTS: Of the 76 cases, MUC1 and MUC5AC immunoreactivities were observed in 49 (64.5%) and in 32 (42.1%) of gastric carcinoma tissues, respectively. MUC1 expression was significantly correlated to the depth of invasion, lymph node metastasis, peritoneal dissemination, and tumor stage. On the other hand, MUC5AC was inversely associated with depth of invasion, lymph node metastasis, liver metastasis, and tumor stage. Multivariate analyses indicated that tumor stage and MUC1 mucin expression were independently correlated with overall survival. The patients with MUC1+/MUC5AC- antigen staining in carcinoma tissues showed the lowest survival rate among four phenotypes. In contrast, the patients with MUC1-/MUC5AC+ antigen staining in carcinoma tissues showed the highest survival rate. CONCLUSIONS: Altogether these data suggest that combined evaluation of MUC1 and MUC5AC mucin staining may be clinically helpful to predict outcome in patients with gastric cancer. PMID- 12884240 TI - Total anterior parietal peritonectomy. PMID- 12884241 TI - Molecular features, regulation, and function of monocarboxylate transporters: implications for drug delivery. AB - The diffusion of monocarboxylates such as lactate and pyruvate across the plasma membrane of mammalian cells is facilitated by a family of integral membrane transport proteins, the monocarboxylate transporters (MCTs). Currently, at least eight unique members of the MCT family have been discovered and orthologs to each have been identified in a variety of species. Four MCTs (MCT1-MCT4) have been functionally characterized. Each isoform possesses unique biochemical properties such as kinetic constants and sensitivity to known MCT inhibitors. Several fold changes in the expression of MCTs may be evoked by altered physiological conditions, yet the molecular mechanisms underlying the regulation of MCTs are poorly understood. Post-translational regulation of MCT1 and MCT4 occurs, in part, by interaction with CD147, an accessory protein that is necessary for trafficking, localization, and functional expression of these transporters. Because of the physiological importance of monocarboxylates to the overall maintenance of metabolic homeostasis, the function of MCTs is significant to several pathologies that occur with disease, such as ischemic stroke and cancer. Finally, the expression of MCT1 in the epithelium of the small intestine and colon and in the blood-brain barrier may provide routes for the intestinal and blood to brain transfer of carboxylated pharmaceutical agents and other exogenous monocarboxylates. PMID- 12884242 TI - Biological, pharmaceutical, and analytical considerations with respect to the transport media used in the absorption screening system, Caco-2. AB - During the evaluation and selection of drug candidates, the Caco-2 cell culture system is commonly used for the determination of intestinal transport characteristics and to anticipate permeability limited drug absorption. Although classic HBSS-like buffered salt solutions are commonly used to perform Caco-2 transport experiments, different shortcomings (e.g., adsorption and low solubility) have been associated with the use of plain aqueous buffers. As transport experiments performed with unoptimized conditions may compromize the value of the Caco-2 model as a permeation screening tool, many efforts have been made to optimize the experimental conditions of Caco-2 transport assays. In this minireview, the hurdles associated with the use of saline aqueous buffers in Caco 2 transport experiments are summarized and the different options, which have been proposed to overcome these issues, are reviewed and discussed. Biologically, pharmaceutically, as well as analytically relevant media affecting the outcome of the transport experiments are described. Unfortunately, up to now, no systematic studies comparing the different experimental conditions have been performed, jeopardizing the possibility to define a (single) optimal solution to overcome the different issues associated with the use of saline aqueous buffers. Based on the reported options it can be proposed to use DMSO (/=12 bases) for the exertion of immune enhancing function upon mammalian cells. Herein we demonstrate that short ISS (5 7 bases), which exhibit no activity on their own, induce IFN-gamma and IFN-alpha secretion from human peripheral blood mononuclear cells when adsorbed to the surface of cationic poly(D,L-lactide-co-glycolide) microparticles (cPLGA). Utilizing this technique, we discovered a minimal ISS sequence for induction of IFN-gamma and IFN-alpha from human cells: 5'-TCGXX-3'. These short ISS/cPLGA formulations targeted PDC in similar fashion to longer ISS ODN, the activity of which does not require (but is enhanced by) cPLGA. PDC stimulated with short ISS/cPLGA responded with enhanced uptake of ISS and elevated production of cytokines, including IFN-alpha. However, ISS-responsive B cells did not respond to short ISS/cPLGA, underlining the plasmacytoid dendritic cell selectivity of this formulation. These results describe a novel technique for formulating active, but very short, ISS oligodeoxynucleotide that allows for the dissection and characterization of minimal immunostimulatory CpG motifs. PMID- 12884287 TI - 4-1BB enhances CD8+ T cell expansion by regulating cell cycle progression through changes in expression of cyclins D and E and cyclin-dependent kinase inhibitor p27kip1. AB - The T cell costimulatory receptor 4-1BB enhances cell cycle progression and proliferation of CD8(+) T cells in both an IL-2-dependent and -independent manner. In these studies, 4-1BB costimulation was shown to increase cyclin D2, D3, and E expression, and concomitantly down-regulate the expression of the cyclin-dependent kinase inhibitor p27(kip1). 4-1BB increases cyclin D2 transcription via mitogen-activated/extracellular signal-regulated kinase-1/2 and LY294002-sensitive phosphatidylinositol 3-kinase (PI3K) signaling pathways. In addition, 4-1BB up-regulates cyclin D2 translation via PI3K/Akt/mammalian target of rapamycin (mTOR) pathways, presumably triggered by IL-2/IL-2 receptor ligation. The enhanced cyclin D2 and D3 expression initiates up-regulation of cyclin E expression and down-regulation of p27(kip1). Our results suggest a role for cyclin D2, D3, and E, and p27(kip1) proteins in the 4-1BB-mediated cell cycle progression of CD8(+) T cells in vivo. PMID- 12884288 TI - Analysis of allelic expression patterns of IL-2, IL-3, IL-4, and IL-13 in human CD4+ T cell clones. AB - The occurrence of monoallelic expression of cytokine genes in single cells has been convincingly demonstrated, but there have been few reports of this phenomenon in T cell clones. Here we describe studies on the expression of alleles of the human genes encoding IL-2, IL-3, IL-4, and IL-13 in human CD4(+) T cell clones. In contrast to the results reported in mouse T cell clones and single human T cells, we found no evidence for the monoallelic expression of the IL-2, IL-3, and IL-13 genes. The gene for IL-4 showed an imbalance in expression from each allele, indicating differential expression of IL-4 alleles within or between IL-4-expressing cells. PMID- 12884289 TI - Caspase-dependent and -independent apoptosis of mast cells induced by withdrawal of IL-3 is prevented by Toll-like receptor 4-mediated lipopolysaccharide stimulation. AB - IL-3-dependent mucosal-like mast cells undergo apoptosis upon withdrawal of IL-3. Generally, the apoptosis is mediated by the activation of caspases and inhibited by addition of the pan-caspase inhibitors z-VAD-FMK or BOC-D-FMK. However, DNA fragmentation, a typical characteristic of apoptosis, is not inhibited by z-VAD FMK or BOC-D-FMK in mast cell apoptosis. In this study, we demonstrate that the apoptosis of mast cells is mediated by both caspase-dependent and -independent mechanisms. The caspase-independent apoptosis is mediated by the translocation of endonuclease G from mitochondria into nuclei. Withdrawal of IL-3 caused down regulation of Bcl-xL, resulting in a drop in mitochondrial membrane transition potential followed by the release of cytochrome c and endonuclease G from mitochondria. However, stimulation of mast cells through Toll-like receptor 4 (TLR4) by lipopolysaccharide prevented mast cell apoptosis by inducing expression of Bcl-xL. Moreover, the activation of mast cells by LPS is enhanced in the presence of IFN-gamma, which up-regulates the expression of cell surface TLR4. Taken together, these observations provide evidence that mast cells play important roles not only in allergic reactions but also in innate immunity recognizing enterobacteria through TLR4, and are regulated differently from allergic inflammation by Th1 cytokines. PMID- 12884290 TI - The mer receptor tyrosine kinase: expression and function suggest a role in innate immunity. AB - The mer receptor tyrosine kinase mediates phagocytosis of apoptotic cells and modulates cytokine production; it is also required for prevention of systemic autoimmune disease. Using a mer-specific antibody, we have confirmed the presence of mer on macrophages and now report its expression on NK cells, NKT cells, and dendritic cells (DC). We found that DC do not require mer for ingestion of apoptotic cells, as DC from mer-deficient mice phagocytose apoptotic cells normally. Mer was observed in splenic sections on cells outside follicular areas, probably representing DC and macrophages. Mer apparently participates in NKT-cell antigen-induced signaling, as NKT cells from mer-deficient mice evinced much lower cytokine production after in vivo alpha-galactosylceramide stimulation; this defect was intrinsic to the mer-deficient NKT cells. Taken together, these studies show mer expression on cells of the innate immune system. Mer, through its binding of lipid antigens, may not only mediate ingestion of apoptotic cells, but also signal events in NK cells, NKT cells, and DC. PMID- 12884291 TI - Oxidative stress stimulates IL-4 and IL-6 production in mast cells by an APE/Ref 1-dependent pathway. AB - Mast cells are exposed to an oxidative environment in the course of allergic and inflammatory reactions. We have examined the effects of H(2)O(2) stimulation in a primary rat basophilic leukemia cell line (RBL-2H3) and compared with IgE dependent stimulation. Like IgE stimulation, H(2)O(2) up-regulates IL-4 and IL-6 gene expression and cytokine secretion, shows a little effect on IL-5 but does not induce IL-10 gene expression. Simultaneous H(2)O(2) treatment and FcepsilonRI triggering of mast cells has additive effects on IL-4 expression. In addition, we show that both stimuli induce the nuclear translocation of APE/Ref-1, a bifunctional enzyme that stimulates the DNA-binding activity of several transcription factors through the reduction of highly reactive cysteines. Conditional inactivation of APE/Ref-1 expression abolishes H(2)O(2)-induced IL-4 and IL-6 gene expression but does not affect that induced by FcepsilonRI stimulation. Our findings indicate that oxidative stress activates the gene expression of a specific cytokine pattern in mast cells through an APE/Ref-1 dependent pathway, which is distinct from the one that is activated by FcepsilonRI stimulation. Nonetheless, H(2)O(2) and FcepsilonRI signalings are additive in augmenting IL-4 production. Most importantly, oxidative stress can induce a pro-type 2 inflammatory response from mast cells that is independent of FcepsilonRI stimulation. PMID- 12884292 TI - Oxidative-stress-induced T lymphocyte hyporesponsiveness is caused by structural modification rather than proteasomal degradation of crucial TCR signaling molecules. AB - In several human pathologies (e.g. cancer, rheumatoid arthritis, AIDS and leprosy) oxidative stress induces T cell hyporesponsiveness. Hyporesponsive T cells often appear to display impaired expression of some (e.g. TCR-zeta, p56(lck) and LAT) but not all (e.g. TCR-alphabeta and CD3-epsilon) crucial TCR proximal signaling molecules but the underlying mechanisms have as yet not been identified. Using an in vitro system for oxidative-stress-induced T cell hyporesponsiveness we here report two sequential effects of oxidative stress on TCR signaling molecules: protein alterations and proteasomal degradation. We have identified the C-terminal part of TCR-zeta and the membrane-proximal domain of p56(lck) as potential targets for modifications induced by reactive oxygen species. Oxidative-stress-exposed proteins were differentially susceptible to proteasomal degradation: whereas modified TCR-zeta was relatively resistant, reactive oxygen species (ROS)-altered LAT and p56(lck) were much more susceptible. Importantly, we found that T cell hyporesponsiveness best correlated with ROS-dependent protein alteration since inhibition of proteasomal degradation did not restore function. Finally, our data provide an explanation for the paradox of reduced TCR-zeta signals combined with unaltered TCR-alphabeta and CD3 epsilon expression levels: the TCR-zeta chain in hyporesponsive T cells is still expressed but no longer detectable by certain mAb recognizing ROS-sensitive epitopes. PMID- 12884293 TI - Vav exchange factor counteracts the HIV-1 Nef-mediated decrease of plasma membrane GM1 and NF-AT activity in T cells. AB - Several findings support the importance of GM1-enriched lipid microdomains of plasma membrane and of Vav, an essential regulator of actin cytoskeletal rearrangement, in the regulation of T cell activation. Moreover, a functional link among lipid microdomains, Vav and the HIV product Nef has been described. These observations suggest that Nef can modify plasma membrane GM1, affecting the behavior of HIV-infected cells towards antigen recognition and Vav towards counteracting such an effect. We observed that Nef expression, either following viral infection or ectopic expression, significantly decreased the level of plasma membrane GM1 in unstimulated T cells. This down-regulation was associated with the inhibition of NF-AT activation, but not with NF-kappaB activation induced by TCR engagement. Dissecting the signaling pathway that regulates NF-AT activation, we found that Nef inhibited exclusively the Ca(2+)/calcineurin cascade, whereas the JNK cascade and AP-1 transcriptional activity were not affected. Our evidence that Vav overexpression counteracted both the Nef-induced decrease of GM1 expression and the inhibition of NF-AT activity, suggests a novel mechanism by which Nef may interfere with TCR-mediated activation through the modulation of intracellular trafficking and clustering of GM1-enriched microdomains at the cell surface. PMID- 12884294 TI - Human NK cells express Fc receptors for IgA which mediate signal transduction and target cell killing. AB - Receptors for the Fc region of IgG (FcgammaRIIIa, FcgammaRIIc) and IgM (FcmicroR) were previously described on NK cells. In this work the expression of Fc receptors for IgA (FcalphaR) on human NK cells and the signaling events were investigated. The FcalphaR was demonstrated by flow cytometry using secretory IgA (sIgA) and anti-human IgA antibody. The percentage of NK cells (CD3( )CD56(+)CD16(+)) expressing FcalphaR ranged between 55.7% and 95.7%, with a mean +/- SD of 75.2+/-11.8. The association constant and the number of (125)I-labeled sIgA ((125)I-sIgA) molecules bound per cell, calculated by Scatchard analysis, were 2 x 10(7) M(-1) and 1.7 x 10(4), respectively. The binding specificity was proved by inhibition experiments. Cold sIgA but not IgA Fab fragments were able to inhibit (125)I-sIgA binding in a concentration-dependent manner. Binding of sIgA to NK cells was neither inhibited by anti-mannose receptor antibody, nor by L-fucose, D-galactose, D-glucose, D-mannose or N-acetyl-D-glucosamine. Pretreatment of NK cells with polymeric IgA inhibited their capacity to kill (51)Cr-labeled K562 target cells by 34.8%, whereas with monomeric IgA only by 13.1%. Ligand-induced clustering of the FcalphaR resulted in activation of tyrosine kinases Lck, Syk and phosphatidylinositol 3-kinase. The present studies support the concept that human NK cells bind preferentially sIgA and polymeric IgA with moderate affinity via FcalphaR, which is different from the FcalphaRI/CD89 and other carbohydrate-recognizing receptors like mannose receptor/CD206. This novel structure mediates signal transduction and cell killing. PMID- 12884295 TI - Sensitivity of IL-5 production to the cAMP-dependent pathway in human T cells is reduced by exogenous IL-2 in a phosphoinositide 3-kinase-dependent way. AB - The cAMP-dependent pathway plays an important role in the regulation of T cell mediated immune responses by inhibition of T cell proliferation, activation and production of Th1-like cytokines. Depending on costimulatory signals and on the activation status of T cells, cAMP also regulates the production of Th2-like cytokines, yet the mechanism is not completely defined. We investigated the effect of costimulation with IL-2 on cAMP-mediated inhibition of IL-5 secretion and the signaling pathways involved in these effects in freshly isolated, alpha CD3/alpha-CD28-stimulated human T lymphocytes. We demonstrate that IL-2 counteracts the cAMP-mediated inhibitory effects on IL-5 secretion by the modulation of phosphoinositide 3-kinase (PI3-K)-dependent signaling. Our results indicate that phosphorylation of cAMP-responsive element-binding protein (CREB) and the activity of the small GTPase Rap1 are unlikely involved in the protective effect of IL-2. Instead, the effect of IL-2 may be mediated by the PI3-K dependent inactivation of the forkhead-related transcription factor FKHR-L1, down regulation of p27(kip) and abrogation of the cAMP-mediated inhibition of activator protein (AP)-1 binding activity. Together, our results indicate that increased IL-2-dependent PI3-K signaling leads to impaired negative feedback control of the production of Th2-type cytokine IL-5 by the cAMP-dependent pathway. PMID- 12884296 TI - TEA regulates local TCR-Jalpha accessibility through histone acetylation. AB - Enhancer alpha-dependent histone acetylation has been proposed as a molecular mechanism underlying the control of accessibility of recombination signal sequences along the TCRalpha locus. Here we show that chromatin acetylation along the first Jalpha segments is under the dependence of the T early alpha element (TEA), located upstream of TCRJalpha locus. The targeted deletion of TEA leads to an absence of histones H3 and H4 tail acetylation, while maintaining histone acetylation in the region spanning downstream Jalpha segments. During thymocyte maturation, TEA-dependent histone acetylation appears at immature single-positive stage, known to represent the stage of ValphaJalpha initiation. TEA-dependent histone acetylation of the most upstream Jalpha segments leads to enhanced DNA accessibility thus optimizing TCRJalpha usage and increasing Ag receptor diversity potential. PMID- 12884297 TI - Activated Akt promotes increased resting T cell size, CD28-independent T cell growth, and development of autoimmunity and lymphoma. AB - The mechanisms that regulate basal T cell size and metabolic activity are uncertain. Since the phosphatidylinositol-3 phosphate kinase (PI3 K) and Akt (PKB) pathway has been shown in model organisms to regulate both cell size and metabolism, we generated transgenic mice expressing a constitutively active form of Akt (myristoylated Akt, mAkt) in T cells. Naive transgenic T cells were enlarged and had increased rates of glycolysis compared to control T cells. In addition, mAkt transgenic T cells resisted death-by-neglect upon in vitro culture. Upon activation, mAkt-transgenic T cells were less dependent than control cells on costimulation through CD28 and could both grow rapidly and secrete cytokines in the absence of CD28 ligation. In addition, transgenic expression of mAkt led to the accumulation of CD4 T cells and B cells with age. Many aged mAkt-transgenic mice also developed autoimmunity with immunoglobulin deposits on kidney glomeruli and displayed increased incidence of lymphoma. Together, these data show that Akt activation is sufficient to increase basal T cell size and metabolism. Enhancement of T cell metabolism by Akt and more rapid CD28-independent T cell growth may contribute to the accumulation of excess immune cells and the development of lymphoma and autoimmunity. PMID- 12884298 TI - Possible involvement of glial cell line-derived neurotrophic factor and its receptor, GFRalpha1, in survival and maturation of thymocytes. AB - The glial cell line-derived neurotrophic factor (GDNF) and its receptors (GFR) play important roles in the promotion of survival and differentiation of central and peripheral neuronal populations. We show that GFRalpha1, a component of GDNF receptor, was expressed in thymocytes at an early stage of thymocyte-development and was involved in the survival of thymocyte precursors. GFRalpha1and GDNF were expressed in thymus, but not in spleen or lymph nodes in adult mice. During embryonic thymocyte development, GFRalpha1 was predominantly expressed on thymocytes from days 14.5 to 16.5 of gestation, and thereafter its expression gradually declined. In adult thymus, GFRalpha1 was expressed only on CD4(-)CD8(-) double-negative (DN) thymocytes, but not on CD4(+)CD8(+) double-positive or single-positive thymocytes. It was strongly expressed on RAG2(-/-) thymocytes arrested at the DN stage, and ist expression was reduced during their differentiation after in vivo anti-CD3 antibody stimulation. Additionally, fetal thymocyte precursors grew in serum-free medium of the fetal thymus organ culture system in the presence of recombinant GDNF (rGDNF), while the cells without rGDNF died. These results suggested that GDNF/GFRalpha1 are involved in the survival of both the nervous system and DN immature thymocytes. PMID- 12884299 TI - CCR3 functional responses are regulated by both CXCR3 and its ligands CXCL9, CXCL10 and CXCL11. AB - The chemokine receptor CXCR3 is predominantly expressed on T lymphocytes, and its agonists CXCL9, CXCL10 and CXCL11 are IFN-gamma-inducible chemokines that promote Th1 responses. In contrast, the CCR3 agonists CCL11, CCL24 and CCL26 are involved in the recruitment of cells such as eosinophils and basophils during Th2 responses. Here, we report that although CCL11, CCL24 and CCL26 are neither agonists nor antagonists of CXCR3, CCL11 binds with high affinity to CXCR3. This suggests that, in vivo, CXCR3 may act as a decoy receptor, sequestering locally produced CCL11. We also demonstrate that the CXCR3 ligands inhibit CCR3-mediated functional responses of both human eosinophils and CCR3 transfectants induced by all three eotaxins, with CXCL11 being the most efficacious antagonist. The examination of CCR3-CCR1 chimeric constructs revealed that CCL11 and CXCL11 share overlapping binding sites contained within the CCR3 extracellular loops, a region that was previously shown to be essential for effective receptor-activation. Hence, eosinophil responses mediated by chemokines acting at CCR3 may be regulated by two distinct mechanisms: the antagonistic effects of CXCR3 ligands and the sequestration of CCL11 by CXCR3-expressing cells. Such interplay may serve to finely tune inflammatory responses in vivo. PMID- 12884300 TI - ME3738 protects from concanavalin A-induced liver failure via an IL-6-dependent mechanism. AB - ME3738 is a new compound that attenuates liver disease in several models of acute and chronic liver inflammation. We used the concanavalin A (Con A) model to elucidate the molecular mechanisms of ME3738 to block liver cell damage. Pretreatment of BALB/c mice with ME3738 prior to Con A injection resulted in a significant reduction in liver injury. The protective effect of ME3738 prior to Con A injection was associated with a reduction in IL-6 serum levels and NF kappaB DNA binding in liver nuclear extracts. However, STAT3 DNA binding was induced via ME3738 prior to Con A injection. Further analysis showed that ME3738 induces IL-6 serum levels and activates STAT3 DNA binding and target gene transcription. The relevance of this finding was assessed in IL-6(-/-) mice. In these animals, ME3738 induced no increase in IL-6 serum expression, and activation of IL-6-dependent pathways was not found. In addition, ME3738 did not protect IL-6(-/-) animals from Con A-induced liver failure, while IL-6 injection was still effective. Therefore, we demonstrate that ME3738 triggers IL-6 expression, which activates pathways that are relevant to protect from Con A induced liver failure. PMID- 12884301 TI - gp49B1 suppresses stem cell factor-induced mast cell activation-secretion and attendant inflammation in vivo. AB - We report that gp49B1, a mast cell membrane receptor with two immunoreceptor tyrosine-based inhibitory motifs (ITIM), constitutively inhibits mast cell activation-secretion induced by stem cell factor (SCF), a tissue-derived cytokine that also regulates mast cell development. The intradermal injection of SCF into the ears of gp49B1 null (gp49B(-/-)) mice elicited approximately 4- and 2.5-fold more degranulating mast cells and tissue swelling caused by edema, respectively, than in gp49B(+/+) mice. SCF did not induce tissue swelling in mast cell deficient mice, and the responsiveness of gp49B(-/-) mice to mast cell-associated amine and lipid mediators was unaltered. When gp49B(+/+) and gp49B(-/-) mice were pretreated with antagonists of the amines, SCF-induced tissue swelling was reduced by >90% and 60%, respectively, and it was reduced by >90% in both genotypes when a cysteinyl leukotriene receptor antagonist was also provided. Hence, the dominant contribution of secretory granule amines to SCF-induced tissue swelling is the result of gp49B1-mediated inhibition of the production of cysteinyl leukotrienes by mast cells. Our findings also provide the first example of an ITIM-bearing receptor that constitutively suppresses inflammation generated in vivo independently of the adaptive immune response by a receptor that signals through intrinsic tyrosine kinase activity rather than immunoreceptor tyrosine based activation motifs. PMID- 12884302 TI - Induction of arthritis by single monoclonal IgG anti-collagen type II antibodies and enhancement of arthritis in mice lacking inhibitory FcgammaRIIB. AB - IgG anti-collagen type II (CII) antibodies (Ab) can induce arthritis in healthy mice. Here we have investigated if single monoclonal IgG anti-CII Ab can induce arthritis in CIA-susceptible DBA/1 mice and if there is an IgG subclass dependency. The involvement of Fc receptors for IgG (FcgammaR) in anti-CII Ab mediated arthritis was also investigated by comparing the clinical outcome in DBA/1 mice to those in FcgammaR-deficient mice. We demonstrate for the first time that single mAb to naive DBA/1 mice can induce persistent arthritis. Histology of the inflamed joints revealed massive cellular infiltrate and cartilage and bone destruction. All IgG subclasses tested (IgG1, IgG2a and IgG2b) were arthritogenic, with the IgG1 and IgG2b isotypes as the dominating arthritogenic Ab. Pathogenicity was dependent on engagement of activating FcgammaR, as FcRgamma deficient mice were completely resistant to Ab-mediated arthritis. The arthritis induced with the IgG1 and IgG2b Ab was also inhibited by FcgammaRIII disruption, whereas arthritis mediated by the IgG2a Ab was not substantially affected. The arthritic response of the IgG1 and IgG2b isotypes, but not of the IgG2a Ab, was further enhanced in mice lacking the inhibitory FcgammaRIIB. These results demonstrate that single IgG anti-CII mAb can induce erosive arthritis and that IgG anti-CII Ab mediate arthritis by engagement of FcgammaR. PMID- 12884303 TI - Essential pathogenic role of endogenous IL-18 in murine diabetes induced by multiple low doses of streptozotocin. Prevention of hyperglycemia and insulitis by a recombinant IL-18-binding protein: Fc construct. AB - IL-18 is a cytokine structurally and functionally related to IL-1 that, in synergy with IL-12, stimulates the synthesis of IFN-gamma from T lymphocytes and natural killer cells. Because IFN-gamma plays a key pathogenic role in the development of murine immunoinflammatory diabetes induced by multiple low doses of streptozotocin (STZ) we investigated the effect of negating the actions of endogenous IL-18 in this model by administering recombinant IL-18-binding protein:Fc (IL-18 bp:Fc). C57BL/6 mice were injected once daily with 40 mg/kg STZ for 5 consecutive days, day 0 being the first day of STZ challenge. Relative to control animals treated in parallel with either PBS or human IgG, mice treated from day -3 to day 7 with daily doses of 150 microg of IL-18 bp:Fc exhibited lower incidence of diabetes and milder insulitis. In contrast, mice that were treated with IL-18 bp:Fc from day 7 to day 14 exhibited clinical and histological signs of STZ-induced diabetes similar to those of control mice treated with IgG. The protective effect of IL-18 bp:Fc was accompanied by modified ex vivo immune responses, in that spleen cells and peritoneal macrophages contained fewer IFN gamma secreting cells and released lower amounts of nitrite (an index of nitric oxide production) and IL-1beta. We conclude that intact IL-18 function is essential for the full diabetogenic effect of low dose STZ in C57BL/6 mice. PMID- 12884304 TI - Cyclic AMP increases endogenous granulocyte colony-stimulating factor formation in monocytes and THP-1 macrophages despite attenuated TNF-alpha formation. AB - The cytokine granulocyte colony-stimulating factor (G-CSF) is in broad clinical use to treat neutropenia, and trials on its use in immunosuppressed conditions and infections are ongoing. To apply G-CSF effectively, it is crucial to understand the regulation and distribution of its endogenous formation. Since G CSF release is mediated, at least in part, by TNF-alpha formation, we investigated whether drugs suppressing TNF-alpha also impair G-CSF production. Surprisingly, G-CSF formation was enhanced in lipopolysaccharide (LPS)-stimulated blood from a pentoxifylline-treated patient. In the presence of dibutyryl-cAMP, forskolin, tolafentrine or 3-isobutyl-1-methylxanthine, LPS-induced G-CSF formation was enhanced in THP-1 cells, primary monocytes and whole blood. Correspondingly,rp-8-bromo-cAMP suppressed LPS-induced G-CSF release. Addition of prostaglandin E(2) enhanced and indomethacin suppressed G-CSF formation. Reporter gene studies showed that dibutyryl-cAMP enhanced LPS-induced G-CSF promoter activity, indicating a transcriptional up-regulation. Furthermore, disruption of a newly identified putative cAMP-responsive element (CRE) in the G-CSF promoter demonstrated the regulatory role for G-CSF gene transcription. In conclusion, endogenous G-CSF formation critically depends on both TNF-alpha and cyclooxygenase products, exerting effects via cAMP and the CRE in the G-CSF promoter. This might have bearing for drug side effects, putative G-CSF mimetics and our understanding of G-CSF immunobiology. PMID- 12884305 TI - TNF-alpha mediates the induction of nitric oxide synthase in macrophages but not in neutrophils in experimental cutaneous leishmaniasis. AB - Leishmania major infection in C57BL/6 mice is controlled by the activation of a Th1 response and nitric oxide (NO) production by macrophages. TNF-alpha is considered one of the most important cytokines involved in this response. In the present study, we investigated the expression of nitric oxide synthase (iNOS) in the inflammatory cells present in the lesion and draining lymph nodes, and the cytokine production by lymph node cells in animals treated with anti-TNF-alpha. Our results demonstrated that mice treated with anti-TNF-alpha presented an increase in the number of parasites and the size of lesion, but they were able to control the infection. The increase in the lesion size correlated to the reduction of iNOS activity in the draining lymph nodes. Furthermore, the anti-TNF alpha treatment also reduced the expression of iNOS in the macrophages, but did not affect the iNOS expression in the neutrophils. The anti-TNF-alpha mAb did not reduce the iNOS expression in IFN-gamma-stimulated L. major infected neutrophils in vitro. Anti-TNF-alpha mAb treatment caused an increase in the production of IFN-gamma and IL-10 by the lymph node cells from infected mice. Consequently, these results suggest that neutrophils do not respond to anti-TNF-alpha treatment and might be a source of NO to control L. major infection under these experimental conditions. PMID- 12884306 TI - Reduced transforming growth factor-beta1-producing T cells in the duodenal mucosa of children with food allergy. AB - Infant food allergies are increasing, and many breast-fed infants now sensitize to maternally-ingested antigens. As low-dose oral tolerance requires generation of suppressor lymphocytes producing TGF-beta1 (Th3 cells), we studied these cells in duodenal biopsies after diagnostic endoscopy. Spontaneous production of Th1, Th2 and Th3 cytokines by duodenal lymphocytes was studied using flow cytometry in 20 children with no eventual clinico-pathological diagnosis (controls), 30 children with multiple food allergy, nine with celiac disease and six with inflammatory enteropathies. Immunohistochemistry and in situ hybridization were used to localize TGF-beta1 protein and mRNA in matched biopsies. We found no significant Th1/Th2 skewing amongst mucosal lymphocytes in allergic children compared to controls, although celiac and inflammatory enteropathy patients showed increased Th1 responses. By contrast, the allergic children showed reduction of TGF-beta1(+) lymphocytes in both epithelial and lamina propria compartments. Reduction of TGF-beta1 expression was also seen in mononuclear cells and epithelium in food allergy by immunohistochemistry and in situ hybridization. The dominant mucosal abnormality in food allergic children was, thus, not Th2 deviation but impaired generation of Th3 cells. As generation of these cells requires innate immune response to enteric bacteria, we suggest that changing infectious exposures may inhibit primary establishment of basic oral tolerance mechanisms. PMID- 12884307 TI - Measurement and modeling of human T cell kinetics. AB - The ability to measure, describe and interpret T cell kinetics is pivotal in understanding normal lymphocyte homeostasis and diseases that affect T cell numbers. Following in vivo labeling of dividing cells with 6,6-D(2)-glucose in eight healthy volunteers, peripheral blood T cells were sorted by CD4, CD8 and CD45 phenotype. Enrichment of deuterium in DNA was measured by gas chromatography mass spectrometry. A novel model of T cell kinetics, allowing for heterogeneity within T cell pools, was used to analyze data on acquisition and loss of label and calculate proliferation and disappearance rates for each subpopulation. Proliferation rates for CD45RO(+)CD8(+) cells and CD45RO(+)CD4(+) cells were 5.1% and 2.7% /day, respectively (equivalent doubling times: 14 and 26 days). CD45RA(+)CD8(+) lymphocytes and CD45RA(+)CD4(+) lymphocytes had slower proliferation rates, 0.5% and 0.6% / day, respectively (doubling time about 4 months). Disappearance rates of labeled cells were similar for all cell types (7% 12% / day) and exceeded corresponding proliferation rates. This disparity may be understood conceptually in terms of either phenotypic heterogeneity (rapid versus slow turnover pools), or history (recently divided cells are more likely to die). The new kinetic model fits the data closely and avoids the need to postulate a large external source of lymphocytes to maintain equilibrium. PMID- 12884308 TI - CRP-ductin, the mouse homologue of gp-340/deleted in malignant brain tumors 1 (DMBT1), binds gram-positive and gram-negative bacteria and interacts with lung surfactant protein D. AB - CRP-ductin is a protein expressed mainly by mucosal epithelial cells in the mouse. Sequence homologies indicate that CRP-ductin is the mouse homologue of human gp-340, a glycoprotein that agglutinates microorganisms and binds the lung mucosal collectin surfactant protein-D (SP-D). Here we report that purified CRP ductin binds human SP-D in a calcium-dependent manner and that the binding is not inhibited by maltose. The same properties have previously been observed for gp 340 binding of SP-D. CRP-ductin also showed calcium-dependent binding to both gram-positive and -negative bacteria. A polyclonal antibody raised against gp-340 reacted specifically with CRP-ductin in Western blots. Immunoreactivity to CRP ductin was found in the exocrine pancreas, in epithelial cells throughout the gastrointestinal tract and in the parotid ducts. A panel of RNA preparations from mouse tissues was screened for CRP-ductin and SP-D expression by reverse transcription-PCR. The pancreas was the main site of synthesis of CRP-ductin, but transcripts were also readily amplified from salivary gland, the gastrointestinal tract, liver, testis, uterus and lung. Lung was the main site of synthesis of SP D, but transcripts were also amplified from uterus, salivary gland, thymus, thyroid gland, pancreas and testis. We conclude that CRP-ductin is the mouse homologue of human gp-340 and that its capacity to bind SP-D as well as gram negative and gram-positive bacteria suggests a role in mucosal immune defense. PMID- 12884311 TI - Effects of rowing on humeral strength. AB - This study used an aggregate measure of robusticity (based on humeral areal and inertial cross-sectional components) to test the hypothesis that rowing on oceans led to more robust humeri than did rowing on rivers or not rowing at all. Results confirmed the hypothesis that male ocean-rowers had more robust humeri than did male river-rowers or nonrowers. However, although the females from the ocean rowing populations did not row, they averaged more robust humeri than did females from the non-ocean-rowing populations. Males averaged greater robusticity than did females. The robusticity of ocean-rowing populations, therefore, does not seem to be due solely to bone remodeling resulting from strenuous rowing. Humeral robusticity is difficult to attribute to specific activities because, in humans, upper limbs are utilized for many activities. Some populations may have more robust humeri because certain environments require greater overall activity levels in order to survive. PMID- 12884309 TI - N-terminal destruction signals lead to rapid degradation of the major histocompatibility complex class II transactivator CIITA. AB - Major histocompatibility complex (MHC) class II molecules play an essential role for the cellular immune response by presenting peptide antigens to CD4(+) T cells. MHC class II molecules and genes show a highly complex expression pattern, which is orchestrated through a master regulatory factor, called CIITA (class II transactivator). CIITA controls MHC class II expression not only qualitatively, but also quantitatively, and has therefore a direct influence on the CD4 T cell dependent immune response. CIITA is itself tightly regulated not only on the transcriptional level, but as we show here also on the protein level. CIITA is subjected to a very rapid protein turnover and shows a half-life of about 30 min. Inhibition of degradation by proteasome inhibitors and the identification of ubiquitylated CIITA intermediates indicate that the degradation of CIITA is mediated by the ubiquitin-proteasome system. We identified two regions mediating degradation within the N-terminal domain of CIITA. N-terminal fusions or deletions stabilized CIITA, indicating that the N termini contribute to degradation. Several non-functional CIITA mutants are partially stabilized, but we provide evidence that transcriptional activity of CIITA is not directly linked to degradation. PMID- 12884312 TI - Beyond palisades: The nature and frequency of late prehistoric deliberate violent trauma in the Chickamauga reservoir of east Tennessee. AB - Based on the presence of palisades and an iconography suggesting a warrior elite, warfare is presumed to be endemic in the Late Mississippian period (AD 1200-1600) of the southeastern United States. Warfare is theorized to play a vital role in the cycling of chiefdoms. However, apart from a few exemplary cases that display double-digit frequencies, very little direct (i.e., skeletal) evidence of violent trauma has dovetailed with the archaeological presumptions of warfare. Eight sites from the Chickamauga Reservoir of east Tennessee were examined for skeletal evidence of deliberate violent trauma. Violent trauma was anticipated because these sites are in close proximity and consist of two adjacent, sociopolitically distinct, and temporally overlapping phases: Dallas (AD 1300-1600) and Mouse Creek (AD 1400-1600). In addition to small, round, nonlethal ectocranial blunt force trauma (BFT) on the frontal and upper parietal bones, inflicted projectile points and scalping were identified. The low total trauma frequency in the Dallas sample (3.86%, n = 259) is consistent with emerging evidence from east and west Tennessee Late Mississippian data, but significantly different from Mouse Creek (8.06%, n = 273). The proportion of nonlethal cranial BFT in the collective Chickamauga sample is large and at odds with the Tennessee River Valley comparative literature. Based on other bioarchaeological literature, this pattern suggests intragroup violence, but not face-to-face ritual contests. It is better explained as interpersonal conflict resolution along codified lines. This is consistent with southeastern ethnohistoric data and may explain the more frequent cranial BFT in the less stratified Mouse Creek phase, which likely would not have had an overarching civil authority. PMID- 12884313 TI - Cranial variation in the Marquesas Islands. AB - The Marquesas Islands have traditionally been divided into a northwestern and a southeastern group, a division which reflects language dialect differences. Additionally, archaeological studies have also suggested that differences in material culture existed between the northwestern and southeastern islands. This study examines Marquesan cranial discrete and metric traits to evaluate the level of intra-archipelago heterogeneity, and to determine if a northwest/southeast division is evident cranially. The data consist of 28 cranial discrete traits and 49 craniofacial measurements of prehistoric Marquesans. Male and female data are pooled for discrete trait and metric data, following a Z-score standardization technique adjustment. The data represent three island samples: Nuku Hiva (northwest), Fatuiva (southeast), and a combined Tahuata/Hiva 'Oa (southeast). Of the 28 discrete traits, 16 are utilized in a mean measure of divergence analysis that provides scores of 0.259 for Fatuiva-Tahuata/Hiva 'Oa, 1.850 for Nuku Hiva Fatuiva, and 1.491 for Nuku Hiva-Tahuata/Hiva 'Oa. Of the 49 craniofacial measurements, 46 are utilized in RMET/NORM analyses, providing unbiased D(2) values of 0.0433 for Fatuiva-Tahuata/Hiva 'Oa, 0.1328 for Nuku Hiva-Fatuiva, and 0.0813 for Nuku Hiva-Tahuata/Hiva 'Oa. The islands of the southeastern group are closer to each other than either was to the island of the northwestern group. When a sample from 'Ua Huka is included in the craniometric analysis, the unbiased D(2) values of 0.0829, 0.1175, and 0.0431 are calculated for 'Ua Huka and Nuku Hiva, and Fatuiva and Tahuata/Hiva 'Oa pairings, respectively, indicating a close similarity of 'Ua Huka to the southeastern islands. Mean measure of divergence analysis of cranial discrete traits as well as RMET/NORM analyses of craniometric variables reveal that differences exist between the islands of the northwestern and southeastern Marquesas Islands. These results support previous research that documented linguistic and cultural differences between these regions of the archipelago. However, the results indicate that 'Ua Huka, an island traditionally included in the northwestern Marquesas Islands, has an affinity to the southeastern Marquesas Islands, possibly due to its pivotal position as a waypoint in the Marquesas Island interaction sphere. PMID- 12884314 TI - Brief communication: age and fractal dimensions of human sagittal and coronal sutures. AB - The fractal dimensions of human sagittal and coronal sutures were calculated on 31 complete skulls from the Terry Collection. The aim was to investigate whether the fractal dimension, relying on the whole sutural length, might yield a better description of age-related changes in sutural morphology, as opposed to other methods of quantification, which generally rely on more arbitrary scoring systems. However, the fractal dimension did not yield better age correlations than other previously described methods. At best, the results reflected the general observation that young adults below age 40 years display an age-related development, but that it is impossible to arrive at any precise age determinations for older adults. It seems that for some individuals, suture obliteration simply does not take place, even at an advanced age, whereas for others, suture obliteration progresses rapidly. Until a better understanding of sutural biology is reached, this will render cranial sutures only marginally useful in age determination. This does not mean, however, that investigations should not be made to elucidate more unbiased methods of sutural morphology quantification. PMID- 12884315 TI - Brief communication: multiplex X/Y-PCR improves sex identification in aDNA analysis. AB - This study introduces a polymerase chain reaction (PCR)-based multiplex approach to improve the certainty of molecular sex identification on archaeological skeletal material. We coamplified amelogenin, two X-chromosomal short tandem repeats (STRs) (DXS6789 and DXS9898), and two Y-specific STRs (DYS391 and DYS392). The amplification results of this multiplex approach back each other up, and enable a reliable sex identification. This coamplification of X- and Y specific markers in a multiplex assay combines the added advantage of positive identification of both female and male individuals with raising the validity of the diagnosis by obtaining multiple data simultaneously. This multiplex system was successfully applied to 3,000-year-old bone material. PMID- 12884316 TI - Ontogenetic characteristics of the vomeronasal organ in Saguinus geoffroyi and Leontopithecus rosalia, with comparisons to other primates. AB - It has been suggested that the variability of the primate vomeronasal organ (VNO) may be greater than previously thought, especially among New World monkeys. It is not clear to what extent VNO variation reflects ontogenetic, functional, or phylogenetic differences among primates. The present study investigated VNO anatomy in an ontogenetic series of two genera of callitrichid primates, in order to assess recent attempts to develop VNO character states and to examine the evidence for VNO functionality at different life stages. A sample of six Leontopithecus rosalia, one L. chrysomelas, and six Saguinus geoffroyi was serially sectioned and stained using various methods. Two adult Callithrix jacchus were also sectioned for comparative purposes. The VNO of each primate was examined by light microscopy along its entire rostrocaudal extent. VNOs of the tamarins were described to determine whether they fit into 1 of 3 character states recently attributed to various New World monkeys. At birth, the two species of tamarins differed in the nature of communication between the VNO and nasopalatine duct (NPD). Two of 3 neonatal S. geoffroyi exhibited a fused VNO duct in a more dorsal position (adjacent to the nasal cavity) compared to that of L. rosalia. The VNO duct communicated with the NPD and was patent in neonatal L. rosalia. Both species appeared to have an age-related increase in the amount of sensory epithelium in the VNO. Subadult L. rosalia had caudal regions of the VNO that were exceptionally well-developed, similar to those of strepsirhine primates. Compared to subadults, all adult callitrichids appeared to have more ventral communications of the VNO duct directly into the NPD. Adult S. geoffroyi and L. chrysomelas both had VNO sensory epithelium separated by multiple patches of nonsensory epithelium. This contrasted with the VNOs of C. jacchus, which had a nearly continuous distribution of receptors on all surfaces of the VNO. The findings indicate that tamarins have delayed maturation of the VNO epithelium, and that some species have little or no perinatal function. These results also suggest that ontogenetic changes in craniofacial form may alter the position of the VNO in tamarins. The present study supports the use of at least two character states to categorize the VNO of various callitrichids, but it is suggested that one of these, previously called "reduced sensory epithelium" should be instead termed "interrupted sensory epithelium." The distribution of VNO sensory epithelium does not appear to reflect phylogenetic influences; it is more likely a functional characteristic that varies throughout postnatal life. Therefore, this chemosensory system has a high degree of plasticity relating to age and function, which in some instances can confound the use of characteristics as phylogenetic traits. Further study is needed to quantify VNO receptors in various species to determine if functional differences exist and if some species have more precocious VNO function than others. PMID- 12884317 TI - Osteon remodeling dynamics in the Cayo Santiago Macaca mulatta: The effect of matriline. AB - At the microstructural level, bones remodel throughout life. This process is recorded in bone cortex as osteons. A more comprehensive understanding of the interaction between genetic regulation and environmental factors in osteon remodeling will increase the value of this skeletal record and enable more accurate reconstruction of individual life histories. The purpose of this study was to examine the contribution of maternal lineage to normal age and sex variation in osteon remodeling dynamics in Macaca mulatta. Femoral cross sections from 57 Cayo Santiago-derived rhesus macaques representing five matrilines were examined to evaluate the effect of genetic relatedness on osteon remodeling dynamics. Analysis of variance revealed an effect of maternal lineage on osteon area and Haversian canal area. The other variables did not differ significantly among matrilines. Analysis of covariance revealed no significant interactions among age, sex, and matriline for any of the microstructural variables. PMID- 12884318 TI - Mitochondrial DNA sequence from an enigmatic gorilla population (Gorilla gorilla uellensis). AB - Although today gorillas are found in only two widely separate, discontinuous western and eastern African populations, rumors of the existence of an additional gorilla population in central Africa have inspired recent unsuccessful field expeditions in search of the "mystery ape" termed Gorilla gorilla uellensis. Such a gorilla population would have considerable conservation and scientific interest, and would presumably have descended from a population of gorillas that was thought to exist until the end of the 19th century on the Uele River in the current-day Democratic Republic of Congo. However, the sole evidence for the existence of these gorillas is three skulls and one mandible brought to the Royal Museum for Central Africa (Tervuren, Belgium) in 1898. We determined a mitochondrial DNA sequence from one of these specimens and compared it to sequences from other gorillas. Contrary to expectations, the sequence obtained did not exhibit the phylogenetic distinctiveness typical of a representative of a peripheral isolated population. Rather, the results suggest a scenario in which the museum specimens did not originally derive from the northern Congo, but were brought from the area of current distribution of western gorillas to that location; the subsequent discovery and collection of the specimens there gave rise to the false inference of a local gorilla population. PMID- 12884319 TI - Asymmetric regional cerebral blood flow in sedated baboons measured by positron emission tomography (PET). AB - The analysis of structural brain asymmetry has been a focal point in anthropological theories of human brain evolution and the development of lateralized behaviors. While physiological brain asymmetries have been documented for humans and animals presenting with pathological conditions or under certain activation tasks, published studies on baseline asymmetries in healthy individuals have produced conflicting results. We tested for the presence of cerebral blood flow asymmetries in 7 healthy, sedated baboons using positron emission tomography, a method of in vivo autoradiography. Five of the 7 baboons exhibited hemispheric asymmetries in which left-sided flow was significantly greater than right-sided flow. Furthermore, the degree of asymmetry in 8 of 24 brain regions was found to be significantly correlated with age; older individuals exhibited a higher degree of asymmetry than younger individuals. Cerebral blood flow itself was uncorrelated with age, and differences between males and females were not significant. PMID- 12884320 TI - Does variation in sample size explain individual differences in hand preferences of chimpanzees (Pan troglodytes)? An empirical study and reply to Palmer (2002). PMID- 12884322 TI - Testing a method: Paleodemography and proportional fitting. PMID- 12884324 TI - On defining the imaging sciences: Tthe first annual Biomedical Imaging Research Opportunities Workshop. PMID- 12884325 TI - Proton MR spectroscopic studies of chronic alcohol exposure on the rat brain. AB - PURPOSE: To better understand the long-term pathophysiologic mechanisms of alcoholism-related organic brain damage by serially assessing brain metabolites in chronically exposed rats using both in vivo magnetic resonance spectroscopy (MRS) and high-resolution nuclear magnetic resonance (NMR) from brain extracts. MATERIALS AND METHODS: The alcoholic regimen was continued up to 60 weeks. In vivo proton MRS studies were performed at 200 MHz using a small animal imaging/spectrometer. In vitro rat brain extracts were also examined using a 500 MHz vertical bore magnet. Comparison measurements were also obtained in an age matched control group. RESULTS: In vivo results showed that there is a significant increase in the Cho/NAA ratio in the chronic alcohol-exposed group that reached a maximum around 16 weeks. After 44 weeks of alcohol exposure, Cho/NAA in the alcohol group decreased significantly from its maximum value to a value that was significantly lower than those from the control groups. Brain extract studies demonstrated that PC and GPC were the main components responsible for the observed in vivo spectral changes after 16 and 60 weeks of alcohol consumption, respectively. CONCLUSION: The fluctuation of choline-containing metabolites during alcohol intoxication could explain sometimes seemingly conflicting and confusing results from MRS studies in human and animal studies in which the duration of alcohol consumption and amount are varied widely. PMID- 12884326 TI - Functional MRI of the rat lumbar spinal cord involving painful stimulation and the effect of peripheral joint mobilization. AB - PURPOSE: To examine neuronal activation in the spinal cord due to secondary hyperalgesia resulting from intrajoint capsaicin injection, and the effect of physiotherapy manipulation, using functional magnetic resonance imaging (fMRI), in alpha-chloralose anesthetized rats. MATERIALS AND METHODS: FMRI of the rat lumbar spinal cord was performed at 9.4 Tesla. Stimuli included injection of 25 microL of capsaicin (128 microg/mL in 7.5% dimethyl sulfoxide [DMSO]) into the right forepaw or 75 microL into the right ankle joint followed by a light touch stimulus, with and without physiotherapy manipulation. RESULTS: Activation of pain areas of the spinal cord (dorsal horn) was found in all animals after injection of capsaicin into the plantar surface of the rat hindpaw and ankle joint. Overlay maps depicting activations and deactivations showed significant reproducibility between experiments. Greater overlay of activations were observed for intrajoint compared to intradermal capsaicin injection. The distribution of activations after stimulation of the hindpaw using a light touch stimulus was somewhat more varied; activation of the dorsal horn was evident, with greater overlap resulting when joint mobilization was not performed. CONCLUSION: Results suggest a trend toward decreased areas of activation in the spinal cord associated with pain, as a result of hyperalgesia, following physiotherapy joint mobilization. PMID- 12884327 TI - Improvement in the quantification of myocardial perfusion using an automatic spline-based registration algorithm. AB - PURPOSE: To improve the quantification of myocardial perfusion by registering the time series of magnetic resonance (MR) images with injection of gadolinium. MATERIALS AND METHODS: Eight patients underwent MR scans to perform myocardial perfusion exam. Two short axis views of the left ventricle (LV) were acquired in free breathing. Two masks for performing the spatial registration of the images were evaluated. The registration was based on pixel intensity in a multi resolution scheme. The efficiency of this correction was evaluated by calculating geometric residual displacement of the LV and by fitting the data to a compartment model fit with two parameters: K1, the blood-to-myocardium transfer coefficient, and Vd, the distribution volume of the contrast media. RESULTS: The registration stage allowed a decrease in the observed motion of the LV from more than 1.98 +/- 0.68 mm to less than 0.56 +/- 0.18 mm (mean +/- SD). Variability obtained in the perfusion analysis decreased from 46 +/- 103% to 5+/- 4% for K1 parameter and from 18 +/- 21% to 5 +/- 5% for Vd parameter. CONCLUSION: As with manual correction, this automatic motion correction leads to accurate perfusion parameters in dynamic cardiac MR imaging after contrast agent injection. This automatic stage requires placing only one mask over one frame of the perfusion study instead of manually shifting each image to fit a reference image of the perfusion study. PMID- 12884328 TI - Two-dimensional flow quantitative MRI of aortic arch blood flow patterns: Effect of age, sex, and presence of carotid atheromatous disease on prevalence of spiral blood flow. AB - PURPOSE: To determine the effect of age, sex, and presence of carotid atheromatous disease on the presence of aortic spiral blood flow pattern using two-dimensional flow quantitative magnetic resonance imaging (MRI). MATERIALS AND METHODS: Sixty subjects (37 women, 23 men) were examined. Prospective phase contrast flow quantitative MRI (1.5 T, Siemens Symphony) sequences in the plane of the aortic arch, and three-dimensional contrast-enhanced MR angiography of the vessels from the aortic arch to the circle of Willis, were performed. Flow quantitative analysis, using circular regions of interest, in the root, apex, and descending aortic arch to determine the presence of a spiral blood flow pattern was undertaken. The results were correlated with the subjects age, sex, and presence of significant carotid arterial disease. RESULTS: A spiral blood flow pattern was seen during diastole in 43 of 50 (86%), 42 of 48 (88%), and in 26 of 28 (93%) subjects in the root, apex, and descending aortic arch, respectively. Spiral flow was seen during systole in 14 of 35 (40%), 20 of 47 (42%), and 11 of 31 (35%) subjects in the root, apex, and descending aortic arch, respectively. There was no clear effect of age or sex on the presence of spiral flow. Carotid disease was associated with a significant reduction in the prevalence of systolic spiral flow from 51%-19% subjects (P < 0.05). CONCLUSION: Spiral flow pattern can be seen in the arch of the aorta in clinical practice using flow quantitative MRI. The prevalence of spiral flow pattern does not appear affected by subject age or sex. Carotid atheromatous disease is associated with a reduction in prevalence of systolic spiral flow pattern in the aortic arch. PMID- 12884329 TI - Automated unwrapping of MR phase images applied to BOLD MR-venography at 3 Tesla. AB - PURPOSE: To improve the diagnostic value of BOLD MR-Venography by removing artifacts related to phase wrapping, particularly in regions of large background susceptibilities at high magnetic field strengths. MATERIALS AND METHODS: High resolution, T(2)(*)-weighted, single echo images were acquired on a 3 T system (Medspec 30/80 Avance, Bruker Medical, Ettlingen, Germany) with a three dimensional, first order velocity compensated gradient echo sequence using a quadrature transmit/receive birdcage head coil. Data of nine healthy subjects and 19 patients were evaluated (age range: 27 to 76 years). To achieve improved MR venograms, a fully automated region-growing phase unwrapping algorithm and subsequent high pass filtering were applied to phase images prior to the computation of minimum intensity projections. RESULTS: Considerably improved visualization of venous structures in regions of large background susceptibility induced field inhomogeneities is demonstrated in healthy subjects and patients with brain tumors. CONCLUSION: BOLD MR venograms are improved compared to previous post-processing algorithms. It is now feasible to obtain high-resolution images also in inhomogeneous regions, which increases the clinical potential of BOLD MR-Venography. PMID- 12884330 TI - Liver metastases from pancreatic adenocarcinoma: MR imaging characteristics. AB - PURPOSE: To describe the magnetic resonance imaging spectrum of appearances of liver metastases from pancreatic ductal adenocarcinoma. MATERIALS AND METHODS: We retrospectively evaluated the MRI exams performed between July 1996 and August 2001 in all patients who had liver metastases from pancreatic adenocarcinoma and histopathologic diagnosis from either the primary pancreatic tumor, liver metastases, or both. Sixteen patients were included in the study. All MR studies were performed at 1.5 T with a standard protocol including T1- and T2-weighted images and serial post-gadolinium spoiled gradient echo (SGE) images. Location, size, number, signal characteristics on T1- and T2-weighted images, and pattern of enhancement on serial gadolinium-enhanced SGE images were assessed. RESULTS: The diameter of metastases ranged from a few millimetres to 4 cm, and 12 patients (75%) had only lesions of 1.5 cm or less. Capsular-based liver metastases were found in 13 patients (81%) and three patients had only capsular-based lesions with a diameter under 1.5 cm. Hypervascular lesions were found in six patients (38%) and hypovascular lesions in 10 patients (62%). Perilesional enhancement was present in 10 patients (62%), with six patients (38%) having ring perilesional enhancement and eight patients (50%) having wedge-shaped perilesional enhancement. CONCLUSION: On MR imaging, hepatic metastases from pancreatic adenocarcinoma show a range of enhancement patterns. Hypervascular metastases are not rare. Capsular based distribution, small diameter, and perilesional enhancement are common features. This retrospective study describes the MR imaging spectrum of appearances of liver metastases from pancreatic adenocarcinoma in patients with histopathologic confirmation of the diagnosis. PMID- 12884331 TI - Hepatocellular carcinoma of diffuse type: MR imaging findings and clinical manifestations. AB - PURPOSE: To assess MR imaging findings and clinical manifestations of diffuse type hepatocellular carcinoma (HCC). MATERIALS AND METHODS: We retrospectively reviewed our experience with diffuse HCC from November 1994 to October 2001. MR imaging findings and clinical features were assessed. RESULTS: Twenty-two consecutive patients with diffuse-type HCC (19 men and three women, age range 16 80 years [mean, 52 years]) were identified in a review of liver MR studies. This represented 13% of all patients with HCC imaged during this time period. Diffuse HCC showed a permeative, infiltrative pattern with ill-defined borders and no evidence of convex margination in all cases. At least 50% of the liver volume was involved with tumor. Diffuse-type HCC showed hypointensity in 15 patients, mixed intensity in three, and isointensity in four on T1-weighted images; heterogeneous hyperintensity in 16 patients; and homogeneous hyperintensity in six on T2 weighted MR images. Diffuse-type HCC showed patchy enhancement in 12 patients, miliary enhancement in nine, and minimal enhancement in one on postcontrast early phase images, and showed heterogeneous wash-out in all patients on postcontrast late-phase images. Proximal portal venous tumor thrombosis was seen in all patients. Serum alpha-fetoprotein (AFP) value was elevated (>10 ng/mL) in 14 of 18 patients, and 13 showed a value greater than 500 ng/mL. The four patients who did not have elevated AFP had tumors which were indistinguishable from those in patients with elevated AFP; they also did not have a distinctive clinical history. CONCLUSION: Diffuse-type HCC was typically seen as an extensive, heterogeneous permeative hepatic tumor, with portal venous tumor thrombosis on MR images in all cases. Early enhancement, observed as patchy in 12 and miliary in nine of 22 patients, was a distinctive imaging feature. Elevated serum AFP value was a common finding; however, 22% had normal values. PMID- 12884332 TI - Biliary motor function in gallstone patients evaluated by fatty-meal MR cholangiography. AB - PURPOSE: To investigate the possibility of evaluating biliary motor function with magnetic resonance cholangiography (MRC). MATERIALS AND METHODS: Twenty patients with gallstones and 30 control subjects were studied using fatty-meal MRC. After baseline MRC, they were encouraged to drink 250 mL of milk and underwent postprandial MRC every 10 minutes for 60 minutes. Postprandial changes in gallbladder volume and the diameter of the common duct were assessed as indicators of gallbladder contractility and biliary obstruction, respectively. Postprandial dilatation at 60 minutes was considered indicative of persistent biliary obstruction. RESULTS: Gallbladder ejection fraction was calculated at 66.0% +/- 12.2% (range, 40.3%-88.6%) in the controls. Gallbladder volume expressed as a percentage of the baseline value was significantly larger at 20-60 minutes in the gallstone patients than in the controls. Gallbladder ejection fraction varied widely (mean, 46.4% +/- 24.4%; range, 2.8%-81.5%) and was significantly reduced in comparison with that of the controls (P < 0.01). In two gallstone patients with co-existing ductal stones, transient postprandial dilatation associated with ampullary impaction was observed. Persistent biliary obstruction was not indicated in any subjects. CONCLUSION: The results of this study suggest the feasibility of fatty-meal MRC, as well as its potential for evaluating biliary motor function. PMID- 12884333 TI - Phantom and animal studies of a new hepatobiliary agent for MR imaging: comparison of Gd-DTPA-DeA with Gd-EOB-DTPA. AB - PURPOSE: To investigate the characteristics of Gd-DTPA-DeA as a hepatobiliary contrast agent for MR imaging in comparison with those of Gd-EOB-DTPA. MATERIALS AND METHODS: We undertook phantom experiments to assess T1 relaxivity for Gd-DTPA DeA, Gd-EOB-DTPA, and Gd-DTPA in human plasma. For Gd-DTPA-DeA and Gd-EOB-DTPA, we evaluated the contrast effect in rats using an SPGR sequence. The contrast ratios of liver and abdominal aorta were measured up to 21 minutes after intravenous administration of the agents. Visualization of the bile duct and renal pelvis was also assessed. RESULTS: In human plasma, T1 relaxivity was similar for Gd-DTPA-DeA and Gd-EOB-DTPA, and higher than those for Gd-DTPA. Whereas the contrast ratio of liver peaked about five minutes after the injection of Gd-EOB-DTPA and was followed by a subsequent decline, a continuous rise was shown for Gd-DTPA-DeA, resulting in a larger maximal contrast effect. Contrast ratios of the abdominal aorta were larger for Gd-DTPA-DeA. Biliary excretion was observed for both agents but occurred earlier with Gd-EOB-DTPA. While renal excretion was shown for all rats three minutes after the injection of Gd-EOB DTPA, it was not observed for Gd-DTPA-DeA. CONCLUSION: Gd-DTPA-DeA may be used as a hepatobiliary contrast agent and shows different pharmacokinetics from Gd-EOB DTPA. PMID- 12884334 TI - Autosomal dominant polycystic kidney disease: MR imaging evaluation using current techniques. AB - PURPOSE: To determine the MR imaging findings of autosomal dominant polycystic kidney disease using current imaging techniques. MATERIALS AND METHODS: We reviewed our five-year experience with MR imaging of autosomal dominant polycystic kidney disease (ADPKD) to determine the spectrum of appearance of kidney disease, the occurrence of cysts in other abdominal organs, the size and number of cysts in the kidneys and other organs, and the association with other benign or malignant disease. Thirty patients (17 men and 13 women, age range 30 to 88 years old) with ADPKD were included in this study. All patients were examined by MR imaging including T2-weighted single-shot echo-train spin-echo and pre- and post-gadolinium chelate spoiled gradient-echo imaging. RESULTS: All kidneys were involved with multiple, varying sized cysts scattered throughout the parenchyma. Giant renal cysts (>8 cm) were associated with pain in the only two patients who possessed them. Hemorrhage in renal cysts was observed in all kidneys with a heterogeneous pattern of involvement on non-contrast T1- and T2 weighted images, reflecting hemorrhage of varying age. The mean kidney size for the right kidney was 17.4 cm in length, 10.3 cm in transverse, and 9.4 cm in antero-posterior diameter (AP); and for the left kidney, 15.9 cm in the length, 9.3 cm in the transverse, and 9.3 cm in AP diameter. Other organs involved included the liver (22 patients), the pancreas (three patients), with two of the above-mentioned patients having both liver and pancreas cysts, and the spleen (one patient) who had both liver and splenic cysts. Massive liver involvement with large cysts was associated with abdominal pain. Malignant disease was present in five patients, including two patients with renal cell carcinoma, one with bladder cancer, one with lung cancer, and one patient with anal adenocarcinoma. Comparison of pre- and post-contrast T1-weighted images was essential to detect renal cancer. CONCLUSION: All kidneys in patients with ADPKD had extensive, varying-sized cysts and in all cases some cysts showed evidence of hemorrhage. The liver was the second most common organ to be involved with cystic disease, in 73% of patients. Large cysts in the kidneys and liver were associated with abdominal pain. PMID- 12884335 TI - Serial MR imaging with MS-325 for evaluating female sexual arousal response: determination of intrasubject reproducibility. AB - PURPOSE: To determine if a similar sexual arousal response in normal, healthy women could be obtained and monitored by serial magnetic resonance (MR) imaging at two separate sessions. MATERIALS AND METHODS: Serial imaging of the external genitalia was performed on nine healthy, sexually functional women at two separate MR sessions after administration of the contrast agent, MS-325. Images were obtained every three minutes during a 45-minute study period during each MR session. The second MR session began approximately 45 minutes after the end of the first MR session. While undergoing imaging, subjects viewed videotapes that contained neutral and sexually-explicit material through an audiovisual system. Analysis performed at each time point consisted of visual evaluation of the images, clitoral and femoral vein signal intensity measurements, relative regional blood volume calculations, and clitoral volume measurements. Statistical analysis of the results consisted of calculating correlation coefficients of the two MR sessions by using the least square fit method. RESULTS: All nine subjects reported sexual arousal on subjective questionnaires at each MR session. Post contrast MS-325 MR images showed strong enhancement of the external genitalia at each session. There was excellent correlation between the two sessions for the clitoral volume measurements of all nine subjects. The correlation coefficient, r(2), was 0.95. CONCLUSION: The sexual arousal response in normal, healthy women can be monitored by serial imaging combined with the use of the contrast agent, MS-325, and similar results can be reproduced at two different MR sessions. This method holds promise for future studies of women with female sexual arousal dysfunction. PMID- 12884336 TI - Fat suppression gradient-echo magnetic resonance imaging of experimental articular cartilage lesions: comparison between phase-contrast method at 0.23T and chemical shift selective method at 1.5T. AB - PURPOSE: To evaluate the diagnostic performance of a newly developed single-scan phase-contrast water-fat imaging technique for fat suppression at 0.23T open magnet, compared to the conventional chemical shift selective fat suppression method at 1.5T, in the detection of experimental articular cartilage lesions. MATERIALS AND METHODS: Sixty regions of 20 knee joint specimens of pigs with artificially created articular cartilage lesions were examined with 0.23T and 1.5T MR scanners. Sagittal fat-suppressed three-dimensional gradient-echo (3D GRE) images, obtained with the phase-contrast method at 0.23T, and fat-suppressed three-dimensional spoiled gradient recalled echo (3D SPGR) images, obtained with a chemical shift selective method at 1.5T, were evaluated. Diagnostic performance was analyzed. The conspicuity of the lesions, the amount of artifacts, and the uniformity of fat suppression were evaluated. The contrast-to-noise (CNR) values of cartilage-to-bone marrow, and cartilage-to-infrapatellar fat were calculated. RESULTS: At 0.23T, sensitivity and specificity were 80% and 95% for partial cartilage lesions (grade 2), and 91% and 100% for full-thickness lesions (grade 3). At 1.5T, sensitivity and specificity were 85% and 95% for grade 2 lesions, and 96% and 97% for grade 3 lesions. No significant difference was detected in the conspicuity of lesions. The uniformity of fat suppression was more constant with 3D SPGR images compared to 3D GRE images. More susceptibility artifacts, derived from the procedure of creating lesions, were detected at 1.5T. The cartilage-to-fat CNRs were significantly higher with high-field images. CONCLUSION: Phase-contrast method for fat suppression at 0.23T is a useful technique in detecting articular cartilage lesions. PMID- 12884337 TI - Improved detection of metastases on magnetic resonance images by digital tissue recognition: validation using VX-2 tumor in the rabbit. AB - PURPOSE: To evaluate the ability of a prototype digital tissue recognition (DTR) system to improve the accuracy of detection of metastases on magnetic resonance (MR) images in the rabbit VX-2 tumor model. MATERIALS AND METHODS: Multiple MR imaging (MRI) sequences, including pre-contrast and post-contrast enhanced T1 weighted, T2-weighted, proton-density, and fast short inversion time inversion recovery (FSTIR), were acquired for six rabbits implanted with VX-2 adenocarcinoma. For each rabbit, DTR used the MR intensity characteristics of a known tumor site to highlight other areas suspicious for tumor. Three independent veterinary radiologists with extensive experience in animal MRI interpreted the images for tumor both without and with the results of DTR. The conventional and DTR-assisted interpretations were compared to pathology. RESULTS: Using DTR, the radiologists found an average of 13.2% more true positive sites with a 10.3% reduction in false positives compared to unassisted interpretation. The improvement for the radiologists was statistically significant (McNemar's test, P = 0.0004). The agreement between radiologists using DTR was consistently higher than for their conventional interpretations (kappa statistic). CONCLUSION: Compared with conventional interpretation of MR images, the use of DTR provided a statistically significant improvement in the accuracy of locating more and smaller sites of tumor. This improvement was achieved without the benefit of post contrast images. PMID- 12884338 TI - A framework for a streamline-based probabilistic index of connectivity (PICo) using a structural interpretation of MRI diffusion measurements. AB - PURPOSE: To establish a general methodology for quantifying streamline-based diffusion fiber tracking methods in terms of probability of connection between points and/or regions. MATERIALS AND METHODS: The commonly used streamline approach is adapted to exploit the uncertainty in the orientation of the principal direction of diffusion defined for each image voxel. Running the streamline process repeatedly using Monte Carlo methods to exploit this inherent uncertainty generates maps of connection probability. Uncertainty is defined by interpreting the shape of the diffusion orientation profile provided by the diffusion tensor in terms of the underlying microstructure. RESULTS: Two candidates for describing the uncertainty in the diffusion tensor are proposed and maps of probability of connection to chosen start points or regions are generated in a number of major tracts. CONCLUSION: The methods presented provide a generic framework for utilizing streamline methods to generate probabilistic maps of connectivity. PMID- 12884339 TI - Acute infarct of the corpus callosum: appearance on diffusion-weighted MR imaging and MR spectroscopy. AB - Few reports of corpus callosum infarction are published in the radiologic literature. We present a case of an atypical infarction of the corpus callosum and adjacent gyri involving both anterior cerebral artery territories. Standard magnetic resonance (MR) imaging was performed including diffusion weighted imaging. In addition, perfusion-weighted imaging and MR spectroscopy were employed to further characterize the patient's corpus callosal lesion. On MR imaging and spectroscopic examination, the lesion demonstrated T2 hyper intensity, restricted diffusion, and the metabolite lactate. While the MR imaging appearance of this bilateral infarction has some characteristics that mimic tumor, employment of the full range of MR techniques can obviate the need for biopsy in acute infarcts of the corpus callosum. PMID- 12884340 TI - Quantitative perfusion mapping of the human lung using 1H spin labeling. AB - PURPOSE: To evaluate the feasibility and reproducibility of a noninvasive, rapid and quantitative pulmonary perfusion mapping method using a two-compartment tissue model in combination with a (1)H spin labeling technique. MATERIALS AND METHODS: Ten healthy volunteers and three patients with cystic fibrosis (CF) were examined on a 1.5-T whole-body scanner. Global and selective lung T(1) maps based on an inversion recovery Snapshot FLASH technique were acquired from each subject with breath-holds at end-expiration. For comparison, corresponding Gd-DTPA enhanced (1)H MR perfusion images were also obtained from each CF patient. RESULTS: Quantitative perfusion maps were calculated from the global and selective T(1) maps. The measured perfusion rates of the upper right lung in volunteers ranged from 400 to 600 mL/100 g/minute. The method showed a high intra study reproducibility and low relative errors. In CF-patients, perfusion defects detected using Gd-DTPA-enhanced MR imaging were also detected using the spin labeling method. The perfusion rates of diseased lung tissues were less than 200 mL/100 g/minute. CONCLUSION: Noninvasive, robust and quantitative (1)H MR mapping of pulmonary perfusion was successfully performed using a rapid lung T(1) mapping in combination with spin labeling within the imaging slice. The proposed method has the potential to provide both important qualitative functional information and quantitative pulmonary perfusion rates in various lung diseases at various stages without the need of contrast agents. PMID- 12884341 TI - Characterization of interpolation effects in cine anatomic and phase-velocity images. AB - PURPOSE: To compare actual and predicted frequency response of reconstruction algorithms applied to anatomic and phase-contrast velocity cine images. MATERIALS AND METHODS: Anatomic and phase-contrast velocity segmented cine gradient echo images were collected from a stationary, doped-agarose phantom, using non-phase contrast cine and phase-contrast cine with one to three directional encodings, one to 16 views per segment, and nearest-neighbor or linear interpolation. Temporal power spectra from object pixels were fit to linear transfer function models; nearest-neighbor interpolation by a sinc function and linear interpolation by a sinc(2) function. RESULTS: Simple linear transfer function models predicted >98% of the observed power spectral variation. Finite word effects produced small systematic differences at high temporal frequency. CONCLUSION: Temporal power spectra of cine images collected from stationary objects completely characterizes low pass filtration effects of interpolation. PMID- 12884343 TI - Comparison of pharyngoesophageal segment pressure in total laryngectomy patients with and without pharyngeal neurectomy. AB - BACKGROUND: To compare pharyngoesophageal segment (PES) pressure values in total laryngectomy patients with and without pharyngeal neurectomy (PN) in the early postoperative period. METHODS: Forty-five previously untreated laryngeal carcinoma patients were enrolled into this prospective randomized study. Twenty of them underwent total laryngectomy with PN, and 25 underwent total laryngectomy without PN. PES pressures were measured on the tenth postoperative day with a four-channel catheter. RESULTS: Average PES pressures in patients with and without pharyngeal neurectomy were 12.82 +/- 6.11 mmHg and 17.40 +/-.72 mmHg respectively (p <.05). When compared with the critical point of 20 mmHg that is closely related to voice attainment in the group without pharyngeal neurectomy, 10 (40%) patients had pressure levels greater than 20 mmHg and in the other group only 1 (5%) patient had a pressure level greater than 20 mmHg. The difference between the groups with pressure levels greater than 20 mmHg was found to be statistically significant (p <.05). CONCLUSIONS: Pharyngeal neurectomy results in a statistically significant decrease of PES pressures in total laryngectomy patients. PMID- 12884344 TI - Treatment of gustatory sweating (Frey's syndrome) with botulinum toxin A. AB - BACKGROUND: Gustatory sweating is a common complication of parotid surgery. Injection of botulinum toxin A has been reported as a safe and effective treatment option for patients with Frey's syndrome. PATIENTS AND METHODS: A total of 69 patients who had undergone superficial parotidectomy because of adenoma were evaluated with respect to the incidence of Frey's syndrome and treatment interest. Minor's iodine starch test was used to detect the affected skin area. Affected skin areas were documented using a digital camera; skin areas were evaluated morphometrically. A single injection of Botox per 1 cm(2) skin field was administered to those patients interested in treatment. RESULTS: Of 43 patients (62%) with gustatory sweating, 33 patients requested treatment. The affected skin area varied from 16 cm(2) to 81 cm(2). The individual Botox dosage ranged from 16 to 80 IU. All relevant clinical symptoms of sweating disappeared within 1 week after a single injection. Treatment was well tolerated with no side effects. CONCLUSION: Botox A injection is a safe and effective treatment with long-lasting effects for patients with extensive gustatory sweating. PMID- 12884345 TI - Distant metastases after definitive radiotherapy for squamous cell carcinoma of the head and neck. AB - PURPOSE: To analyze parameters that influence the risk of distant metastases after definitive radiotherapy. METHODS: Between 1983 and 1997, 873 patients were treated with definitive radiotherapy and had follow-up for 2 years or more. Univariate and multivariate analyses were performed to evaluate risk factors that might influence the risk of distant metastases. RESULTS: The 5-year distant metastasis-free survival rate was 86%. Univariate analyses revealed that the risk of distant metastases was significantly influenced by gender (p =.0092), primary site (p =.0023), T stage (p <.0001), N stage (p <.0001), overall stage (p <.0001), level of nodal metastases in the neck (p <.0001), histologic differentiation (p =.0096), control above the clavicles (p <.0001), and time to locoregional recurrence (p <.0001). Multivariate analysis of freedom from distant metastases revealed that gender (p =.0390), T stage (p <.0001), N stage (p =.0060), nodal level (p <.0001), and locoregional control (p <.0001) significantly influenced this end point. Multivariate analysis revealed that gender (p =.0049), T stage (p <.0001), N stage (p <.0001), and locoregional control (p <.0001) significantly influenced cause-specific survival. CONCLUSIONS: The risk of distant metastases after definitive radiotherapy is 14% at 5 years and is significantly influenced by gender, T stage, N stage, nodal level, and locoregional control. PMID- 12884346 TI - Therapy evaluation of laryngeal carcinomas by tyrosine-pet. AB - INTRODUCTION: One of the major problems in head and neck oncology is determination of tumor status after radiotherapy. Physical examination and conventional imaging by CT and MRI do not always accurately differentiate between residual or recurrent tumor and posttreatment inflammation, fibrosis, edema, or scarring. The feasibility of positron emission tomography (PET) with L-[1-(11)C] tyrosine (TYR) for therapy evaluation of laryngeal squamous cell carcinomas by identification of residual or recurrent disease after radiotherapy was investigated. PATIENTS AND METHODS: Nineteen patients with laryngeal carcinomas had standard workups with endoscopy and conventional imaging. All subjects underwent a TYR PET scan (PET1) before definitive treatment. For determination of tumor status, a second TYR PET scan (PET2) was performed 3 months after radiotherapy. At the time of scanning, seven patients were clinically suspected of having residual disease, and in these cases, additional CT imaging and biopsies during endoscopy were performed. During the minimal follow-up period of 29 months, six patients had clinical suspicion of recurrent disease. In these six cases, a third TYR PET (PET3), CT imaging, and biopsy were performed. RESULTS: All pretreatment tumors were depicted by TYR PET (PET1). Three months after radiotherapy, sensitivity and specificity of TYR PET (PET2) for discrimination between residual tumor and benign posttreatment tissue changes were both 100%, and for CT, 50% and 67%, respectively. For detection of recurrent tumor during follow-up, sensitivity and specificity of TYR PET (PET3) were also 100%, and CT, 75% and 50%, respectively. CONCLUSIONS: Dynamic TYR PET is an accurate imaging modality for therapy evaluation in detection of residual and recurrent disease with higher sensitivity (100%) and specificity (100%) for discrimination of tumor status by TYR PET compared with conventional imaging. PMID- 12884347 TI - Using thallium-201 SPECT to detect recurrent or residual nasopharyngeal carcinoma after radiotherapy in patients with indeterminate CT findings. AB - BACKGROUND: The aim of this study is to evaluate the usefulness of thallium-201 (Tl-201) single-photon emission CT (SPECT) to detect recurrent or residual nasopharyngeal carcinomas (NPC) when CT findings are indeterminate. METHODS: After radiotherapy, 30 NPC patients with indeterminate CT findings were included. CT, Tl-201, and biopsy were performed at least 4 months after radiotherapy and within 1 week of each other. The final results were based on histopathologic findings and clinical follow-up after at least 6 months. RESULTS: The rates for sensitivity, specificity, and accuracy of Tl-201 SPECT in detecting recurrent or residual NPC when there were indeterminate CT findings, were 86.7%, 93.3%, and 90.0%, respectively. CONCLUSIONS: These results suggest that Tl-201 SPECT is effective in detecting recurrent or residual NPC when CT findings are indeterminate. PMID- 12884348 TI - A histological comparison of deep and superficial lobe pleomorphic adenomas of the parotid gland. AB - BACKGROUND: Excision of parotid superficial lobe pleomorphic adenomas requires removal of a surrounding cuff of normal parotid tissue. Less aggressive dissection in removing pleomorphic adenomas that occur in the deep lobe of the parotid gland does not seem to compromise prognosis in these patients. We attempted to define histologic characteristics, differentiating superficial and deep lobe tumors, in an attempt to explain this clinical phenomenon. METHOD: Thirty-one pleomorphic adenomas, 12 deep-lobe tumors, and 19 superficial lobe tumors were analyzed and compared, looking at tumor size, capsule thickness, penetration of tumor through capsule, and predominant cell types present. RESULTS: The superficial lobe tumors had significantly thinner capsules (p =.02). There was increased extracapsular extension of tumor in the superficial lobe group compared with the deep lobe group (79% and 58%, respectively). The tumors were larger in patients with deep lobe lesions (2.6 cm vs 3.6 cm). There was no difference in predominant cell types. CONCLUSIONS: The anatomic location of deep lobe tumors is a likely explanation for the histologic differences observed in this study. These important differences allow less aggressive dissection in deep lobe tumors without compromising prognosis. PMID- 12884349 TI - P53 mutation correlates with cisplatin sensitivity in head and neck squamous cell carcinoma lines. AB - BACKGROUND: A critical factor for successful organ preservation treatment in head and neck cancer may be selecting tumors that respond to chemotherapy and radiation. Previous results in patients indicated that tumors that overexpressed p53 were more sensitive to chemotherapy than those that did not overexpress p53. METHODS: To determine the relationship of p53 mutations to sensitivity to cisplatin in vitro, 23 head and neck squamous cell carcinoma (HNSCC) cell lines were analyzed for cisplatin sensitivity, p53 expression, and p53 mutation status. RESULTS: Mutations of the p53 gene were identified in 13 of 23 of the cell lines tested. Mutation of the p53 gene was significantly associated with high levels of expression of the p53 protein. The average ID(50) (drug dose required to inhibit 50% of cell growth) for cell lines with mutant p53 was 6.8 microM, whereas the average ID(50) for cell lines with wild-type p53 was 13.7 microM. CONCLUSIONS: These in vitro data support a role for mutation of the p53 tumor suppressor gene as a marker for response to cisplatin in HNSCC. PMID- 12884350 TI - Anaplastic transformation of thyroid cancer: review of clinical, pathologic, and molecular evidence provides new insights into disease biology and future therapy. AB - BACKGROUND: Anaplastic thyroid cancer ranks among the most lethal of all human malignancies. Its rarity and rapidly fatal course have made it a difficult cancer to both study and treat. Unfortunately, there has been little progress in the management and control of this malignancy. Anaplastic transformation, or the intratumoral evolution of anaplastic carcinoma from pre-existing differentiated thyroid cancer, has become a well-accepted process, despite a limited understanding of its underlying mechanisms. METHODS: It is through review of the literature that an understanding of the aggressive disease biology can be developed. The aim of this review is to evaluate the relevant clinical, pathologic, and molecular studies to develop an insight into the mechanisms that underlie the intratumoral molecular evolution of anaplastic thyroid carcinoma. RESULTS/CONCLUSION: It is based on an understanding of this process that effective treatments for this aggressive malignancy are currently being developed. PMID- 12884351 TI - Hard palate resection, microvascular reconstruction, and prosthetic restoration: a 14-year retrospective analysis. AB - BACKGROUND: This retrospective analysis was conducted to evaluate self-reported patient and clinician assessed functional outcomes of patients who have undergone ablative hard palate oncologic resection and microvascular free-flap reconstruction with and without maxillofacial prosthetic intervention. METHODS: All Head and Neck Surgery Service, Plastic and Reconstruction Surgery Service, and Dental Service charts and progress notes entered into the Institutional Health Care Information System of 57 patients who underwent hard palate resection and microvascular reconstruction at Memorial Sloan-Kettering Cancer Center (MSKCC) between January 1, 1988, and December 31, 2001 were reviewed retrospectively by three maxillofacial prosthodontists (BJB, JMH, IMZ). Prosthetic and nonprosthetic rehabilitation outcomes for each patient were reported as having normal; limited; or poor esthetics and function according to each prosthodontist's clinical evaluations; patients' self-perceptions; and feedback from friends, family members, and/or spouse. Speech intelligibility was determined in the same manner but reported as normal, hypernasal, and hyponasal. RESULTS: Most of the 26 prosthetically rehabilitated patients were reported as having normal esthetics (81%), normal function (77%), normal speech (96%), and were able to return to per oral full diets (81%) without any restrictions. CONCLUSIONS: Acceptable oral rehabilitation outcomes were reported for most prosthetically rehabilitated free-flap patients. Normal speech can be anticipated with a high degree of certainty, and a high percentage of prosthetically restored free-flap patients achieved per oral diets. PMID- 12884352 TI - Cricopharyngeal spasm and Zenker's diverticulum. AB - Cricopharyngeal spasm and Zenker's diverticulum represent disorders of the pharyngoesophageal junction for which a unifying theory of etiology has yet to be established. There is, however, a large body of evidence that supports an association with gastroesophageal reflux. Cricopharyngeal myotomy is the key to successful management of both disorders. Newer transoral endoscopic techniques of management have a lower overall morbidity than traditional open approaches in appropriately selected patients and are therefore gaining popularity as the preferred method of treatment. PMID- 12884353 TI - Parathyroid adenoma autoinfarction: a report of a case. AB - BACKGROUND: Parathyroid adenoma autoinfarction, although uncommon, is an entity that has been previously reported in the literature; however, the influence of intraoperative parathyroid hormone (PTH) monitoring on therapeutic management has not been reported. METHODS: We present a case of parathyroid autoinfarction that is unique in that it applies a new technology to parathyroid surgery: intraoperative PTH monitoring. RESULTS: Intraoperative PTH monitoring aided in the successful surgical management of this patient. CONCLUSIONS: Intraoperative PTH monitoring can serve as a therapeutic adjunct in the surgical management of parathyroid adenoma autoinfarction. PMID- 12884354 TI - Rhabdomyoma of the head and neck: clinicopathologic features of two cases. AB - BACKGROUND: Extracardiac rhabdomyomas are rare benign tumors of the head and neck. They are frequently misdiagnosed and possibly overtreated when confused with other aggressive tumors. METHODS: This article reports on the clinicopathologic features of two such tumors affecting adult patients and initially seen as slowly-growing, indolent neoplasms. RESULTS: The tumors were of fibrous consistency, mobile, with well-demarcated borders and evident lines of cleavage. Both patients were treated by conservative surgery and remained disease free after prolonged clinical observation. Histologically, both tumors were composed of large eosinophilic cells intermingled with polyhedral clear cells. Both cell types contained abundant glycogen accumulations. CONCLUSIONS: The differential diagnosis is with granular cell tumor and with sarcomas, the former being easily distinguishable morphologically, and the latter be seen with more rapid growth and adherence to the adjacent tissues. On the basis of the preceding features, rhabdomyomas can be suspected at a preoperative stage and adequately managed with enucleation or simple excision. PMID- 12884355 TI - 1H MR spectroscopy of brain tumours and masses. AB - Accurate diagnosis is essential for optimum management and treatment of patients with brain tumours. Proton magnetic resonance spectroscopy ((1)H MRS) provides information non-invasively on tumour biochemistry and has been shown to provide important additional information to that obtained by conventional radiology. We review the current status of (1)H MRS in classifying brain tumour type and grade, for monitoring response to therapy and progression to higher grade, and as a molecular imaging technique for determining tumour extent for treatment planning. PMID- 12884356 TI - Diffusion-weighted MR imaging of the liver of hepatitis C patients. AB - Magnetic resonance diffusion-weighted imaging (DWI) of the liver was investigated to determine whether this method could be used to differentiate between the stages of fibrosis and inflammation for hepatitis C viral infection. DWI data were recorded for 18 hepatitis C patients and 10 control subjects using a modified pulse sequence allowing a 52 ms echo time delay. Acquisitions were performed with breath holding using five different b gradient factor values ranging between 50 and 250 s/mm(2) and in the three axes. Apparent diffusion coefficient (ADC) values were measured from a 5.7 cm(2) area in the central region of the liver. The inflammation and fibrosis grades were evaluated histologically on a biopsy sample. The mean ADC values were 2.30 +/- 1.28 x 10( 3) and 1.79 +/- 0.25 x 10(-3) mm(2)/s for hepatitis C patients and control subjects, respectively. Using our technique, no correlation could be found between the ADC values and the inflammation or fibrosis scores, indicating that tissue changes produced by hepatitis C do not appear to be quantifiable by DWI. PMID- 12884357 TI - Improved spatial localization based on flow-moment-nulled and intra-voxel incoherent motion-weighted fMRI. AB - Functional MRI signal based on the blood oxygenation level-dependent contrast can reveal brain vascular activities secondary to neuronal activation. It could, however, arise from vascular compartments of all sizes, and in particular, be largely influenced by contributions of large vein origins that are distant from the neuronal activities. Alternative contrasts can be generated based on the cerebral blood flow or volume changes that would provide complementary information to help achieve more accurate localization to the small vessel origins. Recent reports also indicated that apparent diffusion coefficient-based contrast using intravoxel incoherent motion (IVIM) weighting could be used to efficiently detect synchronized signal changes with the functional activities. It was found that this contrast has significant arterial contribution where flow changes are more dominant. In this study, a refined approach was proposed that incorporated the flow-moment-nulling (FMN) strategy to study signal changes from the brain activation. The results were then compared with those from conventional IVIM- and BOLD-weighted acquisitions. It was shown that the activated region using the new acquisition strategy had smaller spatial extent, which was contained within the activated areas from the other two methods. Based on the known characteristics of the conventional IVIM and BOLD contrasts, it was inferred that the FMN-IVIM acquisition had improved selective sensitivity towards smaller vessels where volume changes were prevalent. Therefore, such an acquisition method may provide more specific spatial localization closely coupled to the true neuronal activities. PMID- 12884358 TI - Detection of the non-steroidal anti-inflammatory drug niflumic acid in humans: a combined 19F-MRS in vivo and in vitro study. AB - This study describes for the first time results of a (19)F-MRS study on humans exposed to the fluorinated non-steroidal anti-inflammatory drug niflumic acid. The accumulation and elimination of this commercially available selective prostaglandin synthase inhibitor is studied after an oral bolus in the human liver, in blood plasma and in urine samples. The in vivo spectra of the liver display two resonances with a similar increase in signal intensity during the investigation period of 240 min. One resonance refers to the parent compound niflumic acid (P), whereas the second resonance corresponds to a metabolite (M1) formed by the biotransformation by liver enzymes. The spectroscopic comparison with model compounds suggests 4'-hydroxyniflumic acid as the metabolite. During the entire experiment the concentration ratios of these resonances (P/M1) ranged between 0.7 and 0.9, indicating a high metabolite concentration most probably due to an efficient first pass metabolism. Both resonances (P, M1) were observed in the in vitro study of the blood plasma samples after plasma protein denaturation. However, in comparison to the liver spectra, the amount of the metabolite M1 is very small with a P/M1-ratio of 36.6 after 90 min and 16.1 after the end of measurement. This finding suggests an efficient biliary excretion of the metabolite M1, which bypasses the blood circulation system. Both resonances are also identified in the native urine samples. The signal intensity of the parent compound dominates the spectra of all urine samples, whereas the signal intensity of M1 increases slowly reaching a similar value to the parent compound P at the end of the measurement. This observation demonstrates an effective renal elimination of niflumic acid and suggests the existence of an enterohepatic circuit with a re-entry mechanism for the biliary excreted metabolite M1. In the urine spectra, an additional metabolite M2 is found. This resonance exhibits a low but constant signal intensity. The chemical origin of this metabolite is unclear. PMID- 12884359 TI - MR imaging of postischemic neuronal death in the substantia nigra and thalamus following middle cerebral artery occlusion in rats. AB - The goal of this study was to investigate apparent diffusion coefficient (ADC) and T(2) relaxation time (T(2)) in the substantia nigra and thalamus after middle cerebral artery occlusion in rats. In the substantia nigra ipsilateral to infarct, ADC was significantly lower and T(2) was significantly higher on the third and fourth days, but they did not change significantly on the first, second, eighth and 15th days. In the ipsilateral thalamus, ADC and T(2) did not change significantly between the first and fourth days, but were significantly lower on the eighth and 15th days. This combination of MR findings suggested that secondary degeneration in the thalamus was different from that in the substantia nigra. PMID- 12884360 TI - In vivo characterization of fatty acids in human adipose tissue using natural abundance 1H decoupled 13C MRS at 1.5 T: clinical applications to dietary therapy. AB - Natural abundance proton-decoupled (13)C magnetic resonance spectroscopy was used to establish the in vivo lipid composition of normal adipose tissue and the corresponding effects of altered lipid diets. Experiments were performed on a standard 1.5 T clinical MR scanner using a double-tuned (1)H-(13)C coil. Peaks from double-bonded and methylene carbons were analyzed. Normal lipid composition was established in 20 control subjects. For comparison, five subjects on altered lipid diets were studied. Four subjects were on a fish oil supplement diet or predominantly seafood diet (polyunsaturated fatty acids), and one subject was on a Lorenzo's oil diet (monounsaturated fatty acids). Well-resolved (13)C spectra were obtained from the calf adipose tissue with a total acquisition time of 10 min. Model oil solutions were used to identify specific (13)C resonances. Subjects on lipid diets showed significantly elevated levels of monounsaturated and polyunsaturated fatty acids for Lorenzo's and fish oil diets, respectively. We conclude that (13)C MR spectroscopy can readily detect changes in lipid composition due to medium- and long-term therapeutic lipid diets. Since the examination is rapid, robust and noninvasive, opportunities arise for large clinical trials of preventive or therapeutic diets to be performed with (13)C MRS on a clinical MR scanner. PMID- 12884361 TI - Prior knowledge for time domain quantification of in vivo brain or liver 31P MR spectra. AB - Prior knowledge is required when quantifying in vivo (31)P magnetic resonance spectra from the brain or liver. The prior knowledge system we have used models both the phosphomonoester and phosphodiester resonances as two peaks of equal linewidth and fixed relative chemical shift. The analysis of the data is carried out in the time domain, which allows the broad component of the spectra to be modelled. This prior knowledge method has been tested for analysis of in vivo (31)P MR spectra from the liver and brain and gives results consistent with other methods that are also used to analyse the spectra, but with reduced variability. This technique may be utilized for studies requiring serial MR spectroscopy examinations, before and after patient treatment. PMID- 12884363 TI - Ets protein Elf-1 bidirectionally suppresses transcriptional activities of the tumor suppressor Tsc2 gene and the repair-related Nth1 gene. AB - Alterations in the rat tuberous sclerosis gene (Tsc2) cause renal cell carcinomas (RCCs) with complete penetrance. In this study, it was shown that the minimal core promoters of the rat Tsc2 and endonuclease III 1 (Nth1) genes, lying in a 5' to-5' arrangement, were localized in a 0.11-kb region containing two Ets binding sites (EBSs). This region worked as a bidirectional promoter in a single reporter plasmid. Mutational inactivation of each of the two EBSs significantly reduced promoter activity. Moreover, gel shift assays revealed the presence of specific EBSs-protein complexes. These results demonstrate that some members of the Ets family positively regulate the promoter activities of the Tsc2/Nth1 genes by binding to the EBSs. We identified Elf-1 as a binding factor for EBSs through super-shift assays, and detected approximately 35 kDa bands with an EBSs containing DNA probe by Southwestern blot analysis. Forced expression of Elf-1 in cells, however, bidirectionally suppressed the activities of the Tsc2/Nth1 promoters. Elf-1 may be a negative regulator of Tsc2/Nth1 gene expression and may compete against positive regulators for binding to the EBSs. Our observations suggest that mechanisms that inactivate Tsc2 gene expression, such as promoter suppression, may exist. PMID- 12884364 TI - Prostate-specific antitumor activity by probasin promoter-directed p202 expression. AB - p202, an interferon (IFN) inducible protein, arrests cell cycle at G1 phase leading to cell growth retardation. We previously showed that ectopic expression of p202 in human prostate cancer cells renders growth inhibition and suppression of transformation phenotype in vitro. In this report, we showed that prostate cancer cells with stable expression of p202 were less tumorigenic than the parental cells. The antitumor activity of p202 was further demonstrated by an ex vivo treatment of prostate cancer cells with p202 expression vector that showed significant tumor suppression in mouse xenograft model. Importantly, to achieve a prostate-specific antitumor effect by p202, we employed a prostate-specific probasin (ARR2PB) gene promoter to direct p202 expression (ARR2PB-p202) in an androgen receptor (AR)-positive manner. The ARR2PB-p202/liposome complex was systemically administered into mice bearing orthotopic AR-positive prostate tumors. We showed that parenteral administration of an ARR2PB-p202/liposome preparation led to prostate-specific p202 expression and tumor suppression in orthotopic prostate cancer xenograft model. Furthermore, with DNA array technique, we showed that the expression of p202 was accompanied by downregulation of G2/M phase cell-cycle regulators, cyclin B, and p55cdc. Together, our results suggest that p202 suppresses prostate tumor growth, and that a prostate-specific antitumor effect can be achieved by systemic administration of liposome-mediated delivery of ARR2PB-p202. PMID- 12884365 TI - Pifithrin-alpha promotes p53-mediated apoptosis in JB6 cells. AB - Recently, blockage of p53-dependent transcriptional activation and apoptosis by pifithrin-alpha (PFTalpha) has been reported to be useful for reducing the side effects of cancer therapy and the compound is thus thought to be a specific inhibitor of p53 [Komarov et al., Science 1999;285:1733-1737]. Here, we found that PFTalpha did not inhibit UVB- or doxorubicin (Dox)-stimulated p53-mediated transcriptional activation and apoptosis in JB6 cells. Instead, p53-dependent activation and apoptosis were not only induced by PFTalpha itself but were also enhanced by a combination of PFTalpha with UVB or Dox. Furthermore, PFTalpha induced apoptosis was mediated through p53-dependent and -independent signaling pathways. Extracellular signal-regulated kinases and p38 kinase, but not c-jun N terminal kinases (JNKs), were activated, and these activations were required for phosphorylation and accumulation of p53 in the cellular apoptotic response to PFTalpha. Thus, we conclude that PFTalpha is not a specific p53 inhibitor in JB6 cells but is a potential activator of p53-mediated signaling and apoptosis. PMID- 12884366 TI - Elevated basal reactive oxygen species and phospho-Akt in murine keratinocytes resistant to ultraviolet B-induced apoptosis. AB - Resistance to apoptosis may be a critical phenotype that cells must acquire during skin carcinogenesis. The Akt kinase is a known upstream regulator of apoptosis in many cell types and has been shown to be activated by increased reactive oxygen species (ROS). We have previously demonstrated that two malignant variants (6M90 and 6R90) of the mouse keratinocyte 308 cell line have elevated ROS because of loss of catalase activity, and that this elevated ROS confers a growth advantage. We report here that in addition to a growth advantage, chronically increased ROS in the variants resulted in an increase in resistance to ultraviolet (UV) B-induced apoptosis. This resistance was due to basal increases of Akt phosphorylation in the malignant variants compared to the 308 cells. Modulation of ROS in 6M90 and 6R90 cells by catalase overexpression or antioxidant treatment resulted in decreased levels of Akt phosphorylation and subsequent loss of resistance to UVB-induced apoptosis. Conversely, treatment of 308 cells with hydrogen peroxide caused increases in Akt phosphorylation and increased apoptosis resistance. These results indicate that the chronically elevated ROS often observed in tumors may contribute to a malignant phenotype by keeping Akt in a phosphorylated state, resulting in increased apoptosis resistance. PMID- 12884367 TI - Functional evaluation of novel single nucleotide polymorphisms and haplotypes in the promoter regions of CYP1B1 and CYP1A1 genes. AB - Interindividual variation in the expression of the carcinogen- and estrogen metabolizing enzymes cytochrome P4501B1 and 1A1 (CYP1B1 and CYP1A1) has been detected in human lung. To search for polymorphisms with functional consequences for CYP1B1 and CYP1A1 gene expression, we examined 1.5 kb of the promoter region of each gene. Genomic DNA from 21 Caucasian individuals was amplified by polymerase chain reaction (PCR) for direct cycle sequencing. Eight single nucleotide polymorphisms (SNPs) for CYP1B1 and 13 SNPs for CYP1A1 were found. The majority of polymorphisms occurred as multiSNP combinations for individual subjects. The wild-type sequences were cloned into a luciferase reporter construct. The most frequent polymorphisms were then recreated by iterative site directed mutagenesis, replicating single polymorphisms and multiSNP combinations. These wild-type and variant constructs were functionally evaluated in transient transfection experiments employing exposures to either the index polycyclic aromatic hydrocarbon (PAH) inducer benzo[a]pyrene (B[a]P), a composite mixture of cigarette smoke extract (CSE), or the repressor chemopreventive agent trans-3,4,5 trihydroxystilbene (reseveratrol). Results indicated that all wild-type and variant constructs responded in qualitatively concordant fashion to the inducers and to the repressor. The CYP1B1 haplotypes and the majority of CYP1A1 haplotypes were shown to have no functional consequence, as compared to those of the wild type promoter sequences. Two constructs of composite polymorphisms of CYP1A1 appeared to result in a statistically significant increase in basal promoter activity (1.38- and 1.50-fold, respectively), but the degree of functional impact was judged unlikely to be biologically important in vivo. We conclude that the observed promoter region polymorphisms in these genes are common, but are of unclear functional consequence. PMID- 12884369 TI - Comparative performances of selected chiral HPLC, SFC, and CE systems with a chemically diverse sample set. AB - Pharmaceutical companies have a continuous need to resolve new racemates. Analysis may be required in aqueous and nonaqueous media, or in the presence of several different sets of potentially interfering compounds. There is often a preparative requirement. For these reasons analysts may require a number of different separation systems capable of resolving a given pair of enantiomers. We wished to improve upon existing approaches that address this situation and undertook a program of work to screen over 100 racemates, selected for their chemical diversity, on over 100 different chiral HPLC, SFC, and CE systems. Here we report results of this comparison and illustrate the use of rapid gradient screening as a valuable tool for chiral method development. PMID- 12884370 TI - Gas chromatographic separation of saturated aliphatic hydrocarbon enantiomers on permethylated beta-cyclodextrin. AB - Enantiomers of chiral aliphatic hydrocarbons are generally difficult to separate because they lack functional groups to be derivatized in order to generate diastereomers. The systematic and quantitative separation of a series of branched hydrocarbon enantiomers using a chiral cyclodextrin stationary phase and a cryostat-controlled gas chromatograph is described. The use of a cryogenic system allows the improvement of separations for various chiral aliphatic hydrocarbons. The molecular cyclodextrin-based mechanism of the achieved enantiomeric separations is discussed briefly. Possible applications of this analytical technique are summarized, with special emphasis on the planned enantiomeric separation experiment on a cometary nucleus. PMID- 12884371 TI - Novel cinchona carbamate selectors with complementary enantioseparation characteristics for N-acylated amino acids. AB - The synthesis and chromatographic evaluation of the enantiomer separation capabilities of covalently immobilized calix[4]arene-cinchona carbamate hybrid type receptors derived from quinine (QN) and its corresponding C9-epimer (eQN) in different solvents are reported. The receptors display complementary enantiomer separation profiles in terms of elution order, chiral substrate specificity, and mobile phase characteristics, indicating the existence of two distinct chiral recognition mechanisms. The QN-derived receptor binds the (S)-enantiomers of N acylated amino acids more strongly, shows preferential recognition of open chained amino acids, and superior enantioselectivity in polar media such as methanol/acetic acid. In contrast, the eQN congener preferentially recognizes the corresponding (R)-enantiomers, displays good enantioselectivity (alpha up to 1.74) for cyclic amino acids, and enhanced stereodiscriminating properties in apolar mobile phases, e.g., chloroform/acetic acid. A comparison of the enantiomer separation profiles with those of the corresponding QN and eQN tert butyl carbamate congeners indicates no significant level of cooperativity between the calix[4]arene module and the cinchona units in terms of overall chiral recognition, most probably as a consequence of residual conformational flexibility of the calixarene module and the carbamate linkage. PMID- 12884372 TI - Design and synthesis of fluorescent beta-cyclodextrins for the enantioselective sensing of alpha-amino acids. AB - Fluorescent monofunctionalized beta-cyclodextrins bearing a copper(II) binding side arm and a dansyl group (CD-NH-AA-CH(2)CH(2)NH-DNS) were designed as enantioselective sensors for unmodified alpha-amino acids. The side arm was derived from amino acid synthons (AA = L- and D-phenylalanine (1 and 2), L- and D phenylglycine (3 and 4), L-proline (5), and L-cyclohexylglycine (6)) and was chosen in order to contain an amide, an amine, and a sulphonamide group. Enantioselectivity was evaluated by addition of copper(II) complexes of D- or L valine and D- or L-proline. Chiral discrimination in the fluorescence response was observed in all cases, due to a ligand exchange process. The best conditions for these experiments were found to be the use of an excess (10:1) of the copper complex. The cyclodextrin 4 containing a D-phenylglycine unit was found to be poorly enantioselective, as found for 2, suggesting that the best design can be obtained by using L-amino acids. All L-amino acid containing cyclodextrins showed good enantioselectivities, some of which were higher than those already reported for 1. Other analytes related to amino acids were studied using cyclodextrins 1 and 3. Enantiomers of alpha,alpha-disubstituted amino acids, N-methylamino acids, and amino acid amides were found to be discriminated, while beta-phenylalanine and other molecules bearing a poor anchoring group at the alpha-carbon gave poor enantioselectivity. On the basis of the present data a model for the recognition process, based on the formation of ternary diastereomeric complexes, is proposed. PMID- 12884373 TI - Chiroptical detection during liquid chromatography: Deconvolution of overlapping peaks of enantiomers and its applications. AB - The peaks of enantiomers in liquid chromatography (LC) frequently overlap for different reasons. The experimental curve can be deconvolved, i.e., transformed into the two curves of the enantiomers, without any assumption concerning their peak shapes. Besides the usual photometric UV detection, resulting in absorbance A, polarimetric or circular dichroic detection is required, providing the rotation 90 degree angle alpha or the differential absorbance DeltaA, respectively. The accuracy of the ratio alpha/A or DeltaA/A for the pure enantiomers is essential for the quality of the deconvolution. The determination of these ratios, using the overlapping peaks, and the subsequent computer deconvolution of the latter are discussed in more detail than in the earlier publications, e.g. Ref. 1 concerning this particular method. The computer program developed for this purpose is characterized. A condition is given which limits the availability of ratios and, therefore, the possibility of deconvolution. Several novel examples are described which stem from the following fields of application of deconvolved peaks: actual optical purities during LC (on-line analysis), overall optical purity of a sample, purities of chromatographic peaks, and, finally, enantiomerization during LC. PMID- 12884374 TI - Molecular engineering of extended chiral and enantiopure multiporphyrinic architectures: Towards the control of their physico-chemical properties. AB - We report our recent progress in the synthesis and study of extended chiral and enantiopure multiporphyrinic devices. We devoted particular efforts to the study of the influence of both structure and conformation of a molecule on its physicochemical properties, and in this perspective peptides and nucleosides have been chosen as interchromophore linkers. Concerned with the elaboration of molecular wires, we synthesized enantiopure peptides bearing pendant porphyrins. The synthesis of a peptide built according to a predetermined sequence of L lysine derivative holding porphyrin-functionalized lateral chain allowed, beyond the preparation of homo-octaporphyrins with free-base or Zn(II) porphyrins, the elaboration of a pentaporphyrin with an assigned sequence of chromophores. A two dimensional system, a star-shaped pentaporphyrin with chiral enantiopure linkers derived from uridine, was prepared. Electrochemical and photophysical investigations have shown a duality of the physicochemical properties of the core porphyrin. PMID- 12884375 TI - Determination of absolute configuration using ab initio calculation of optical rotation. AB - Ab initio Density Functional Theory (DFT) calculations of transparent spectral region, discrete frequency specific rotations were used to assign the absolute configurations (ACs) of: 1, 2H-naphtho[1,8-bc]thiophene 1-oxide; 2, m-F-phenyl glycidic acid methyl ester; 3, o-Br-phenyl glycidic acid methyl ester; 4, p-CH(3) phenyl glycidic acid methyl ester; 5, 2-(1-hydroxyethyl)-chromen-4-one; and 6, 6 Br-2-(1-hydroxyethyl)-chromen-4-one. The ACs of 5 and 6 were previously determined via X-ray crystallography to be: 5, R(-)/S(+); 6, R(+)/S(-). The ACs obtained using [alpha](D) are the same for both 5 and 6: R(+)/S(-). We conclude that the previously reported AC of 5 is incorrect. PMID- 12884376 TI - Probing the discriminating power of chiral crown hosts by CD spectroscopy. AB - The discriminating efficiency of pyridino- (1), pyridono- and thiopyridono- (2), phenazino- (3), and acridino- (4) 18-crown-6 hosts as well as pyridino- (1) and phenazino- (3) 18-crown-6 hosts with allylic moieties attached either to the macrocyclic ring [X=CH-CH(2)-CH=CH(2) or C(CH(2)-CH=CH(2))(2)] or to the heterocyclic subunit was probed by circular dichroism (CD) spectroscopy using enantiomers of alpha-(1-naphthyl)- ethylamine hydrogen perchlorate (1-NEA). The CD spectra of the diastereomeric complexes as well as the difference and sum of the spectra were analyzed. Titration experiments were also performed monitored by CD or UV spectroscopy. The CD- and (1)H NMR-based enantiomeric preferences were compared. CD spectroscopy showed that the relative stability of heterochiral [(R,R)-host/(S)-guest or (S,S)-host/(R)-guest] complexes generally exceeds that of homochiral [(R,R)/(R)-or (S,S)/(S)] complexes. Bulkier substituents (R=iBu, sBu or tBu) decrease complex stability but increase the discriminating power of the host. According to (1)H NMR titration, attachment of one allylic linker group in position X of the macro ring [(S,S)-1b, (S,S)-1e] changes the discriminating preference (from heterochiral to homochiral) of the parent hosts (S,S)-1a and (S,S)-1d. This change of discriminating preference was not reflected in the CD spectra. Host (S,S)-1g with an allyloxy linker group in position gamma of the pyridine ring gives CD spectra which clearly reflect the high discriminating power and enantiomeric preference of the host. The exciton-coupled circular dichroism (ECCD) spectra of the 1-NEA complexes of phenazino and acridino hosts allow reliable determination of the enantiomeric preference by comparing the A values of the exciton couplets. An allylic group appended to the macrocyclic ring or the N-containing heterocyclic subunit allows for the attachment of the chiral discriminator to the solid matrix of a chromatographic sorbent and also serves as a spectroscopic label of the host. CD spectroscopy is a simple and rapid method, providing qualitative information on enantiomeric discrimination. It can be of great help in designing and testing new host molecules. PMID- 12884377 TI - Synthesis and NMR configurational analysis of 1,3-imidazolidin-4-ones derived from (-)-(S)-phenylethylamine. AB - The preparation and NMR study in the solution state of (-)-(S)-1-acetyl-3 (phenylethyl)-1,3-imidazolidin-4-one, (-)-(2R,1'S)- and (+)-(2S,1'S)-1-acetyl-2 tert-butyl-3-(phenylethyl)-1,3-imidazolidin-4-one, and (-)-(2S,1'S)- and (+) (2R,1'S)-1-carboethoxy-2-tert-butyl-3-(phenylethyl)-1,3-imidazolidin-4-one are described. The orientations of acetyl, carboethoxy, and phenethyl groups attached to nitrogen atoms N-1 and N-2 were deduced from the proton and (13)C chemical shifts. The typical orientation of phenethyl group, which has the hydrogen H-6 syn and close to the oxygen of carbonyl C-4, was observed in (-)-(S)-1-acetyl-3 (phenylethyl)-1,3-imidazolidin-4-one but not in the other four compounds; nevertheless, interpretation of chemical shifts based on the ring current effects correlated with the true configuration of the new stereogenic center at C-2. Four of these imidazolidinones were analyzed by X-ray diffraction to corroborate the configuration determined by NMR. The structures determined by X-ray diffraction revealed the existence of intramolecular hydrogen bonds capable to control the conformation of the phenethyl and acyl groups in the solution state. PMID- 12884378 TI - Hydrogenation of cinchona alkaloids over supported Pt catalyst. AB - The heterogeneous catalytic hydrogenation of two isomeric cinchona alkaloids, cinchonidine and cinchonine, was studied over Pt/Al(2)O(3) in 1N H(2)SO(4) solution under 100 bar H(2) at 25 degrees C. Cinchonidine was transformed into two diastereomeric hexahydroderivatives by hydrogenation of ring A (with N) of the quinoline moiety (yield over 95%, diastereomeric ratio 2/3), whereas hydrogenation of cinchonine resulted in the formation of three products, the major one being formed by the hydrogenation of ring B (without N) (yield 60%). The isolated hexahydroderivatives were investigated by (1)H-, (13)C-NMR, (1)H (1)H COSY, (1)H-(13)C HetCOSY, and NOESY spectroscopy and were used as modifiers in the heterogeneous enantioselective hydrogenation of ethyl pyruvate and as catalysts in the Michael addition of ethyl 2-oxocyclopentanecarboxylate to methyl vinyl ketone. PMID- 12884379 TI - Enantioselective direct aldol addition of acetone to aliphatic aldehydes. AB - The asymmetric direct aldol addition of acetone to aliphatic aldehydes catalyzed by D-proline, L-proline, and its derivatives was studied. While excellent results could be obtained in neat acetone using alpha-branched aldehydes, unbranched and beta-branched aldehydes gave moderate results. Two dipeptide derivatives, L-Pro-L Try-CH(2)OH and L-Pro-L-Trp-OCH(3), were prepared and tested in this reaction and both were found to be able to induce enantioselectivities. The ee-values in the case of some aldehydes approached that obtained with L-proline. Immobilization of L-proline on a polystyrene resin by its carboxylic group provided a catalyst which is able to induce enantioselectivity, can be easily removed from the reaction mixture, and reused without a significant decrease in the enantioselectivity of the beta-hydroxyketones obtained in the cross-aldol additions. PMID- 12884380 TI - Novel P,N-bidentate phosphite ligands in asymmetric catalysis. AB - Results achieved by the authors in the synthesis of chiral P,N-phosphite ligands are summarized. Three groups of new chiral P,N-phosphites are discussed, namely, ligands derived from 1,1'-bi-2-naphthol, ligands possessing an acyclic phosphorus center, and P*-chiral ligands derived from (S)-2-anilinomethylpyrrolidine. An overview of complexation of the ligands with Rh(I) and Pd(II) precursors is given. Accessibility and stability of chiral phosphite ligands possessing acyclic phosphorus was analyzed for the first time along with their efficiency in terms of stereoselectivity. The title ligands are shown to be highly efficient in the Pd-catalyzed allylic alkylation (up to 85% ee) and, especially, allylic sulfonylation (up to 97% ee) reactions in certain cases outperformed all known catalytic systems. PMID- 12884381 TI - Stereoselective synthesis of 1-aminoalkanephosphonic acids with two chiral centers and their activity towards leucine aminopeptidase. AB - The stereoselective synthesis of 1-amino-2-alkylalkanephosphonic acids, namely, compounds bearing two chiral centers, was achieved by the condensation of hypophosphorous acid salts of (R)(+) or (S)(-)-N-alpha-methylbenzylamine with the appropriate aldehydes in isopropanol. Simultaneous deprotection and oxidation by the action of bromine water provided equimolar mixtures of the RS:RR and SR:SS diastereomers of desired acids. They appeared to act as moderate inhibitors of kidney leucine aminopeptidase with potency dependent on the absolute configuration of both centers of chirality. PMID- 12884382 TI - Development of homochiral peptides in the chemical evolutionary process: Separation of homochiral and heterochiral oligopeptides. AB - Living organisms have one-handed structures of L-amino acids in proteins and D sugars in nucleic acids. Although the origins of each one-handed structure (or homochirality) have been discussed for many years, these discussions have been restricted to monomeric compounds, such as amino acids and monosaccharides, or their stereospecific condensation reactions. Oligomers of these compounds have to be considered in the accumulation processes of homochirality because of the differences in physical properties of the diastereomers. High-performance liquid chromatography (HPLC) and the calculation of the partition coefficient values showed that the peptides having heterochiral sequences like L-Ala-D-Ala or D-Ala L-Ala were more hydrophobic than the peptides having homochiral ones (L-Ala-L-Ala and D-Ala-D-Ala). Similar results were given from the calculation of most linear dipeptides and all cyclic ones composed of Gly, Ala, Val, or Asp. In addition, longer homo-oligopeptides composed of Ala, Val, or Asp also gave similar results. This general tendency would be useful for the separation of diastereomeric oligopeptides in water. The results also suggest that the separation of the homochiral peptides from the heterochiral ones by their solubility in water could have progressed in a primitive hydrosphere. PMID- 12884383 TI - Insect chemistry and chirality. AB - Examination of the chemistry of a number of Australian insect species provided examples of unusual structures and encouraged determinations of their absolute stereochemistry by stereocontrolled syntheses and chromatographic comparisons. Inter alia, studies with the fruit-spotting bug (Amblypelta nitida), certain parasitic wasps (Biosteres sp.), the aposematic shield bug (Cantao parentum), and various species of scarab grubs are summarized. The determination of enantiomeric excesses (ee's) for component epoxides, lactones, spiroacetals, and allenes are described. Stereochemical and related aspects of the biosynthesis of spiroacetals in certain fruit-fly species (Bactrocerae sp.) are also presented. PMID- 12884384 TI - Facing chirality in the 21st century: Approaching the challenges in the pharmaceutical industry. AB - How is process R&D organized and operated in today's pharmaceutical industry at the dawn of the 21st century? A way to respond to the challenges with regard to reduced time to market is to build on early involvement and a front-loading approach. This means that activities are initiated during the lead optimization phase starting up to 2 years ahead of candidate drug nomination and a model built on this concept covering the stages through to commercial launch is advocated as the appropriate way forward. However, given the high attrition rate in a pharma R&D pipeline focused risk management needs to be applied and options judiciously evaluated. From a molecular perspective, the chemical targets in many instances present a formidable complexity both with regard to the overall structure but increasingly also when it comes to their stereochemical features. Thus, a novel triazolo pyrimidine compound with six stereogenic centers requiring 28 transformations for its assembly is examined to underscore this, but also the difficulties in designing a feasible route for the relatively simple (S) azetidinecarboxylic acid are highlighted. Furthermore, the successful development of a unique and highly efficient catalytic asymmetric sulfide oxidation to the corresponding (S)-sulfoxide esomeprazole is discussed, together with the remarkable effect that normal sea sand has on the stereoselectivity of a steroid trans-acetalization. PMID- 12884385 TI - Intrinsic rotation and molecular structure. AB - The intrinsic rotation, limiting value of specific rotation at zero concentration, of (R)-(-)-epichlorohydrin was measured in four different solvents, CH(3)OH, CH(2)Cl(2), CHCl(3) and CCl(4). It was found that the sign of rotation in CH(3)OH, and CH(2)Cl(2) solvents is opposite to that in CCl(4). The intrinsic rotation in CHCl(3) is close to zero. This observed pattern was explained using density functional calculations of specific rotation using very large basis sets. It was found that the g-I and g-II conformations of epichlorohydrin have nearly the same magnitude of specific rotation but with opposite sign. When these two conformations have equal populations, as in CHCl(3) solvent, the net rotation is close to zero. When g-II conformation dominates, as in CCl(4) solvent, the observed sign of rotation will be opposite to that when g I conformation dominates, as in CH(2)Cl(2) and CH(3)OH solvents. A combination of intrinsic rotation measurement with density functional prediction of specific rotation is demonstrated to be a practical method for determining the structures of molecules. PMID- 12884386 TI - Calix[4]arene, calix[4]resorcarene, and cyclodextrin derivatives and their lanthanide complexes as chiral NMR shift reagents. AB - Calix[4]arenes, calix[4]resorcarenes, and anionic cyclodextrin derivatives were examined as chiral NMR solvating agents. The calix[4]arenes were prepared by attachment of amino acids through the hydroxyl groups of the phenol rings. Chloroform-, methanol-, and water-soluble derivatives were prepared and tested with a range of substrates. Chloroform-soluble chiral calix[4]resorcarenes were prepared by attachment of chiral primary and secondary amines and examined in NMR applications with a variety of substrates. Sulfated and carboxymethylated beta cyclodextrin are effective at causing enantiomeric discrimination in the (1)H NMR spectra of organic cations. Lanthanide ions associate at the carboxymethyl groups and cause sizeable shifts and enhancements in enantiomeric discrimination in the spectra of organic cations. The enhancements caused by the lanthanide ion are large enough that much lower concentrations of the cyclodextrin can be used as compared to conventional analyses. PMID- 12884387 TI - Some examples of mass spectrometry research in Italy. PMID- 12884388 TI - Mass spectrometry in grape and wine chemistry. Part I: polyphenols. AB - Mass spectrometry, had and still has, a very important role for research and quality control in the viticulture and enology field, and its analytical power is relevant for structural studies on aroma and polyphenolic compounds. Polyphenols are responsible for the taste and color of wine, and confer astringency and structure to the beverage. The knowledge of the anthocyanic structure is very important to predict the aging attitude of wine, and to attempt to resolve problems about color stability. Moreover, polyphenols are the main compounds related to the benefits of wine consumption in the diet, because of their properties in the treatment of circulatory disorders such as capillary fragility, peripheral chronic venous insufficiency, and microangiopathy of the retina. Liquid Chromatography-Mass Spectrometry (LC-MS) techniques are nowadays the best analytical approach to study polyphenols in grape extracts and wine, and are the most effective tool in the study of the structure of anthocyanins. The MS/MS approach is a very powerful tool that permits anthocyanin aglycone and sugar moiety characterization. LC-MS allows the characterization of complex structures of grape polyphenols, such as procyanidins, proanthocyanidins, prodelphinidins, and tannins, and provides experimental evidence for structures that were previously only hypothesized. The matrix-assisted-laser-desorption-ionization time-of-flight (MALDI-TOF) technique is suitable to determine the presence of molecules of higher molecular weight with high accuracy, and it has been applied with success to study procyanidin oligomers up to heptamers in the reflectron mode, and up to nonamers in the linear mode. The levels of resveratrol in wine, an important polyphenol well-known for its beneficial effects, have been determined by SPME and LC-MS, and the former approach led to the best results in terms of sensitivity. PMID- 12884389 TI - Atmospherically relevant ion chemistry of ozone and its cation. AB - The importance of ionic processes that occur in terrestrial, planetary, and stellar atmospheres is receiving increasing recognition. Actually, ions play important, often crucial, roles in a variety of atmospheric processes throughout the universe, and a strong link with the neutral chemistry is also apparent. In the terrestrial atmosphere, the ionic reactions are most relevant in those transient and fleeting events, e.g., lightning, coronas (in thunderstorm clouds and along power lines), where the local ion density is much higher than in unperturbed air, and the chemical systems are typically far from equilibrium. In such cases, ozone, a key molecule for the terrestrial atmosphere, is also present in high local concentrations; it is formed from O(2) by the same transient event. Accordingly, this review provides a survey of the positive ion chemistry of ozone with several of the most important "atmospheric" species: the reactions, the products, and the importance of the examined processes are discussed also in the light of the local thermodynamic disequilibrium (LTD) approach to the chemistry of transient atmospheric events. In all such studies, mass spectrometry is traditionally, and remains today, the experimental technique of choice. The novel application of mass spectrometry to the study of neutral species (NRMS), highly successful for the preparation and positive detection of long-sought, otherwise inaccessible, short-lived neutrals, makes mass spectrometry the most powerful tool now available for the study of the species and processes that are relevant to atmospheric chemistry. Selected examples of the interlink between the neutral and the ionic chemistry are also illustrated. PMID- 12884390 TI - Assessment of protein expression by means of 2-D gel electrophoresis with and without mass spectrometry. AB - Careful examination of current literature, particularly over the last 5 years, reveals a wide range of approaches for the relative quantification of protein expression in cells, tissues, and body fluids. In view of such an observation, it is reasonable to ask whether researchers need new methods, or whether it is more productive to optimize and tune already existing ones. It is generally agreed that none of the existing methodologies on its own can give a full account of protein expression in a complex medium; this limitation, however, has not prevented the use of existing methods to provide valuable information on a wide range of proteins, where their expression has been correlated to certain pathologies and/or to pharmacological, genetic, or environmental factors. In the present work, an attempt is made to review the application of one of these methodologies, namely two-dimensional polyacrylamide gel electrophoresis on its own or in conjunction with mass spectrometry, to assess protein expression, particularly when such expression can be correlated to certain pathologies. PMID- 12884392 TI - Bioanalytical considerations for compounds containing free sulfhydryl groups. AB - Thiol-containing compounds pose bioanalytical challenge in several dimensions due to extreme reactivity of the sulfhydryl group. The development of robust bioanalytical methodology for thiol groups should address the aspects of adequate stabilization in the biological matrix and selectivity considerations. In this context, availability of plethora of thiol reagents provides ample opportunity to achieve the above goals. However, several considerations need to be factored into the decision-making of a suitable scheme for the thiol drugs under investigation. This review provides some critical insights to many such considerations that may be vital for development and validation of methods for thiol-containing drugs. PMID- 12884393 TI - Liquid chromatographic separation of the enantiomers of beta-amino acids on a ligand exchange chiral stationary phase. AB - A liquid chromatographic ligand exchange chiral stationary phase (CSP) derived from (S)-leucinol was applied in the separation of the enantiomers of 12 beta amino acids. The resolution was quite successful especially for the enantiomers of beta-amino acids containing aromatic functional group in the side chain. The chromatographic resolution behaviors were dependent on the organic modifier and Cu(II) concentration in aqueous mobile phase and the column temperature. PMID- 12884394 TI - The benefit of the retrofitting of a conventional LC system to micro LC: a practical evaluation in the field of bioanalysis with fluorimetric detection. AB - The interests in liquid micro-chromatography (higher column efficiencies, increase in sensitivity) are now well established. The enhancement of fluorimetric response induced by the reduction of the inner diameter of columns (4.6, 3.0, 1.0 and 0.3 mm respectively) coupled with adapted detection cells to control the loss of efficiency (8 micro L for the two first columns and 100 nL for the two smaller ones) has been studied in the bioanalytical field, using the plasma determination of native fluorescent antibacterial agents: fluoroquinolones. Ten-fold enhancement of the signal can easily be obtained when substituting a 0.3 mm i.d. column and 100 nL detection cell for a 4.6 mm i.d. column, and 8 micro L detection cell. In addition to inner diameter reduction, the detection cell geometry appears to be an essential parameter to obtain the best enhancement of the recorded signal. Hence, the enhancement of signal with micro-chromatography with fluorimetric detection appears to be a compromise between column inner diameter and flow cell volume reduction. PMID- 12884395 TI - Analysis of three effective components in Fructus corni and its preparations by micellar electrokinetic capillary chromatography. AB - An efficient micellar electrokinetic capillary chromatography method was developed to analyze three major active components including morroniside, loganin and gallic acid in Fructus corni and its six preparations for the first time. The factors that could affect the separation were studied, such as the pH of the buffer, concentrations of SDS, organic modifier and beta-CD, and the applied voltage. The optimum analysis conditions were 10 mmol/L NaH(2)PO(4)-5 mmol/L Na(2)B(4)O(7) (pH 6.8) buffer containing 140 mmol/L SDS, 1 mmol/L beta-CD, 5% (v/v) methanol and 12.5 kV applied voltage. The linearity between the peak-areas and the concentrations of the analytes were investigated, and they exhibit excellent linear behavior over the concentration ranges (correlation coefficients 0.9953-0.9995). In addition, the pK(a) of gallic acid was determined using capillary zone electrophoresis. The result was consistent with that reported by the literature. PMID- 12884396 TI - Preliminary study on application of impregnated synthetic peptide TLC stationary phases for the pre-screening of 5-HT1A ligands. AB - Chromatographic parameters (deltaR(f)), defined as a difference in the migration of tested compound on the control and peptide impregnated silica gel TLC plates, were determined for 42 arylpiperazine derivatives. An amino acid sequence of the peptide used for impregnation was derived from the III transmembrane segment of the 5-HT(1A) receptor in the close vicinity of aspartic acid (Asp 166) residue. It was found that the deltaR(f) values obtained in a model employing tetrapeptide P4LA (ADVL), as well as the calculated logP correlate with 5-HT(1A) receptor affinity of the studied compounds. PMID- 12884397 TI - Estimation of the lipophilicity of antiarrhythmic and antihypertensive active 1 substituted pyrrolidin-2-one and pyrrolidine derivatives. AB - The lipophilicity of some antiarrhythmic and antihypertensive active 1-[2-hydroxy or 1-[2-acetoxy-3-(4-aryl-1-piperazinyl)propyl]pyrrolidin-2-one derivatives (1 12) has been investigated. Their lipophilicity (R(MO) and log k') was determined by reversed-phase thin-layer chromatography and reversed-phase high-performance liquid chromatography with mixtures of acetonitrile and Tris buffer as mobile phases. The partition coefficients of compounds 1-12 (log P(ScilogP)) were also calculated with the ScilogP program. Comparison of R(MO), log k' and calculated log D(7.0 ScilogP) values enabled calculation of clog D(7.0 TLC) and clog D(7.0 HPLC) values. Preliminary quantitative structure-activity relationship studies indicated that for active compounds there is a dependence between affinity for alpha(2)-adrenoceptors and their clog D(7.0 HPLC) values. PMID- 12884398 TI - Thin layer chromatographic resolution of some 2-arylpropionic acid enantiomers using L-(-)-serine, L-(-)-threonine and a mixture of L-(-)-serine and L-(-) threonine-impregnated silica gel as stationary phases. AB - Impregnated silica TLC plates with L-(-)-serine and L-(-)-threonine and a mixture of L-(-)-serine and L-(-)-threonine (1:1) as chiral selectors were prepared to use as chiral stationary phases (CSPs) in thin layer chromatography. The resolution of the enantiomers of 2-arylpropionic drugs, including ibuprofen, ibuproxam, ketoprofen, pranoprofen, benoxaprofen, flurbiprofen and tiaprofenic acid was investigated on these CSPs. A mobile phase system of acetonitrile methanol-water (16:4:0.5, v/v/v) was used. The spots were detected with iodine vapours and the detection limits were found to range between 0.25 and 0.5 micro g/mL for all racemic compounds investigated. The effect of temperature, pH and concentration of the impregnating chiral selectors on resolution has been studied. PMID- 12884399 TI - Monitoring manufacturing process yields, purity and stability of structural variants of PEGylated staphylokinase mutant SY161 by quantitative reverse-phase chromatography. AB - Staphylokinase variant SY161 is a recombinant mutant of the Staphylococcus aureus polypeptide staphylokinase (Sak), and is currently in human clinical trials as a thrombolytic agent. The 15 kDa single chain SY161 protein is expressed as a soluble cytoplasmic product in E. coli with a single cysteine inserted near the N terminus. The protein as extracted from E. coli is a mixture of both monomeric and intermolecularly disulfide crosslinked species. To improve protein purification yields SY161 is sulfitolyzed during the early stages of production, preventing disulfide formation. The protein is later modified during manufacturing to incorporate a single 5 kDa polyethylene glycol group on the single sulfhydryl sidechain. We have developed and qualified a reverse-phase chromatographic method to quantitate SY161 during product manufacturing. We discuss the use of the assay during manufacturing development to monitor fermentation yields, the SY161 PEGylation reaction, and as an in-process manufacturing control assay. The assay has been applied as a product purity and identity release assay and is suitable for use in assessing product structural integrity during stability testing. The assay has a linear range of quantitation for SY161 from at least 0.15 to 16 micro g, and is-in addition capable of detecting and quantitating protein de-PEGylation events and host cell-derived protein contaminants. PMID- 12884400 TI - An optimized method for determination of intracellular glutathione in mouse macrophage cultures by fluorimetric high-performance liquid chromatography. AB - We have optimized a method for the determination of intracellular glutathione by high-performance liquid chromatography, using fluorimetric detection. To minimize artifacts and provide an accurate determination of intracellular glutathione, cell extracts were prepared using extraction conditions specifically designed to inhibit autoxidation and enzymatic degradation of glutathione. The sensitivity of the method was enhanced by adjusting the dansyl chloride derivatization reaction with regard to parameters such as pH, reaction time and dansyl chloride concentration. Both oxidized and reduced forms of glutathione were quantified using the refined method in extracts of oxidatively stressed J774A.1 mouse macrophage cells and reflected an expected shift in cellular redox status. PMID- 12884401 TI - Methods for the identification of chemical respiratory allergens in rodents: comparisons of cytokine profiling with induced changes in serum IgE. AB - No validated or widely recognized test methods are currently available for the prospective identification of chemicals with the potential to cause respiratory allergy. The cellular and molecular mechanisms that result in the induction of chemical sensitization of the respiratory tract are unclear, although there is evidence for the selective development of T helper 2 (Th2)-type responses and, in some cases, the production of IgE antibody. We have therefore examined the utility of cytokine profiling using BALB/c mice, together with the measurement of induced increases in the total serum concentration of IgE in the Brown Norway (BN) rat, as markers for the prospective identification of chemical respiratory allergens. Responses provoked by the reference respiratory allergen trimellitic anhydride (TMA) have been compared with those stimulated by the respiratory sensitizing diisocyanates toluene diisocyanate (TDI) and hexamethylene diisocyanate (HDI) and by the acid anhydride hexahydrophthalic anhydride (HHPA). Topical exposure of BN rats to TMA, TDI and HHPA each provoked marked immune activation (increases in lymph node cellularity and proliferation). However, only treatment with TMA stimulated vigorous increases in the total serum concentration of IgE. In contrast, exposure to HHPA, TDI or HDI failed to provoke significant changes in serum IgE concentration or induced only transient and relatively weak increases in serum IgE levels. In parallel experiments using BALB/c strain mice, however, topical application of all four chemical respiratory allergens provoked a marked Th2-type cytokine secretion profile in draining lymph node cells. These data suggest that the measurement of induced changes in serum IgE is not sufficiently sensitive for the robust identification of chemical respiratory allergens. Furthermore, irrespective of the reasons for variations in TMA-induced IgE production among BN rats, doubts remain regarding the utility of these animals for the characterization of immune responses to chemical allergens. Cytokine profiling using the BALB/c strain mouse apparently provides a more robust method for the hazard assessment of chemical respiratory allergens. PMID- 12884402 TI - Evidence for redox cycling of lawsone (2-hydroxy-1,4-naphthoquinone) in the presence of the hypoxanthine/xanthine oxidase system. AB - This study reports that lawsone (2-hydroxy-1,4-naphthoquinone) undergoes redox cycling in the presence of the hypoxanthine/xanthine oxidase system. The rate of cytochrome c reduction obtained in the presence of 80 microM lawsone was almost three times the rate of cytochrome c reduction measured in its absence. This increase in the rate of cytochrome c reduction was partially inhibited by superoxide dismutase, suggesting the involvement of O(2)(.-) in this process. It is remarkable to note that, even though lawsone is considered to be a non-redox cycling quinone in vitro, this quinone was shown to be more toxic in vivo in rats than menadione, causing haemolytic anemia of an oxidative nature and renal damage. The view that this quinone is a non-redox-cycling quinone was based on the inability of one-electron-transferring flavoenzymes such as NADPH-cytochrome c reductase to reduce this naphthoquinone. Our finding that lawsone, like menadione, undergoes redox cycling in the presence of the hypoxanthine/xanthine oxidase system could explain the observed oxidative damage of tissues inflicted by this quinone in rats in vivo. Such an observation therefore reconciles the in vivo toxicity results of this naphthoquinone with those of in vitro experiments. PMID- 12884403 TI - Tissue histopathology, clinical chemistry and behaviour of adult Comt-gene disrupted mice. AB - Catechol-O-methyltransferase (COMT) enzyme is a widely distributed enzyme that catalyses O-methylation of catecholamines and other compounds having a catechol structure. Because there has been some concern about the consequences of a low COMT activity in the development of oestrogen-dependent cancers and because one of the COMT inhibitors, tolcapone, has caused serious liver injuries in Parkinsonian patients, the histopathology and clinical chemistry of Comt-gene disrupted mice were studied at the age of 12 months. Owing to the high COMT activities in liver and kidney and the role of COMT in the metabolism of catechol oestrogens, special emphasis was given to the histology of the liver, kidney and oestrogen-dependent organs such as mammary glands and uterus. The mice of both heterozygous and homozygous genotypes appear to be physically healthy and fertile. Diurnal motility rhythm and behaviour in measuring anxiety and depression were equal in all genotypes. At the age of 12 months, the body weight of homozygous mice was 7-9% lower than that of the other groups. This was reflected in histology as a diminished incidence of vacuolation of liver cells (fatty change). Macroscopic pathology and histopathology revealed no abnormal findings in any COMT genotype. The values of some clinical chemistry parameters, such as alkaline phosphatase, alanine aminotransferase, urea, glucose, calcium and proteins, were at a higher level in homozygous animals compared with the wild type mice. However, all the values remained within the normal physiological range, and the differences in enzyme levels between genotypes were not reflected as histopathological findings in the relevant organs. No changes in haematological parameters or plasma catecholamine concentrations were noted but plasma 3,4-dihydroxyphenylethylene glycol levels were high in COMT null mice. The results suggest that the full or 50% lack of Comt gene as such is not associated with any toxic consequences. PMID- 12884404 TI - Apoptosis induced by different doses of caffeine on Chinese hamster ovary cells. AB - Caffeine has been investigated for its potential mutagenic activity to bacteria, fungi and mammalian cells in culture, and at high concentrations it is also an inducer of apoptosis. Caffeine can exert acute cellular toxicity, including inhibition of cell growth and cell death, in Chinese hamster ovary cells. The aim of this study was to evaluate the cell survival and apoptotic or non-apoptotic effects of caffeine to different concentrations in Chinese hamster ovary cells (CHO-K1). These effects were evaluated by measuring cell viability, caspase 8 activity and fragmented DNA. This study suggests that the concentration of caffeine is of critical importance because high doses of caffeine induce apoptosis and low concentrations can act as an antioxidant. Previously, the cytotoxicity of caffeine was evaluated using a wide range of concentrations by the neutral red test. From this screening, adequate doses were selected to perform the caspase activity and fragmentation DNA studies. The potential antioxidant effect of caffeine was studied using tert-butyl-hydroperoxide as a free-radical generator. The repeatability was checked through three separate tests with the same concentration. PMID- 12884405 TI - Degradation of VX and sulfur mustard by enzymatic haloperoxidation. AB - Chloroperoxidase (CPO) isolated from Caldariomyces fumago (20 U ml(-1)) together with urea hydrogenperoxide (UPER, 0.5 mM) and sodium chloride as co-substrate (NaCl, 0.5 M) caused rapid breakdown of VX (10 microM) (t((1/2)) = 8 s, 25 C, 50 mM tartarate, pH 2.75). Glucose oxidase (GOX, Aspergillus niger) and glucose were used as an alternative source for H(2)O(2). A mixture of GOX (20 U ml(-1)), glucose (GLU 0.45 M), CPO (20 U ml(-1)) and NaCl (0.5 M) caused a 3.8-fold slower degradation of VX (10 microM) (t((1/2)) = 30 s, 25 C, 50 mM tartarate, pH 2.75). The concentrations of H(2)O(2) and chlorine produced by this enzyme/substrate mixture depended mainly on the GLU concentration. Horseradish peroxidase (HRP) together with UPER (1 mM) and sodium iodide (NaI, 0.05 M) caused progressive degradation of VX that was more than 400-fold slower than with CPO (20 U ml(-1)), UPER (0.5 mM) and NaCl (0.5 M) (t((1/2)) = 55 min, 25 C, pH 8). Skin decontamination of VX by CPO was tested in pig-ear skin in vitro. The chemical agent VX (0.01 M, 100 microl) was degraded by 98% within 3 h of skin diffusion when a mixture of UPER/NaCl/CPO was applied 60 min prior to VX application. A mixture of UPER/NaCl without CPO also caused significant VX degradation (94%) during skin diffusion whereas it did not cause any VX degradation in solution. Degradation of VX in skin, obtained without exogenous CPO, may indicate involvement of endogenous intradermal haloperoxidase-like enzyme. Reagent UPER (1 mM) did not cause any degradation of VX in solution or during its skin diffusion. Furthermore, a mixture of CPO, UPER and NaCl caused rapid degradation of sulfur mustard (HD). Sulfur mustard (50 microM) incubated in the presence of CPO (4 U ml(-1)), UPER (0.05 M) and NaCl (0.5 M) at pH 2.75 and 30 C was oxidized by 97% and 99% within 5 and 10 min, respectively. The oxidation products HD sulfoxide, HD sulfone and HD sulfoxidevinyl were identified by GC/MS in the enzymatic chloroperoxidation mixture. PMID- 12884406 TI - Leydig cell death in rats exposed to bismuth subnitrate. AB - In a previous study, rats were treated with one injection of 500 mg kg(-1) bismuth subnitrate intraperitoneally and maintained for 2 weeks. Bismuth was observed in Leydig cells, with a subsequent reduction in serum testosterone levels. In the present study, stereological procedures were used to estimate the number of Leydig cells in the right testis from the retained rats used in the previously published study. The mean number of Leydig cells in the control group was estimated to be 18.7 x 10(6) (coefficient of variation = 0.11), which is comparable to previous estimations. In the group exposed to bismuth the mean was 15.5 x 10(6) (coefficient of variation = 0.18). The observed 17% difference between the two groups was statistically significant (2P = 0.019). The inter individual variation was largest in the bismuth-exposed group. Testis weight and body weight were not significantly reduced after bismuth exposure. These findings support the hypothesis that bismuth has a direct toxic effect on rat Leydig cells and underscores the potential risks of bismuth on male reproduction. PMID- 12884407 TI - Iron status in manganese alloy production workers. AB - The aim of this study was to investigate markers of iron status in production workers with current and long-term exposure to manganese (Mn) alloys. A total of 100 Mn-exposed male workers were compared with 100 male controls matched for age in a cross-sectional study. The geometric mean urinary Mn concentration in the exposed workers was 0.9 nmol mmol(-1) creatinine (range = 0.1-126.3), compared with 0.4 nmol mmol(-1) creatinine (range = 0.1-13.1) in the controls. The index group had been exposed to Mn for 20 years on average (range = 2.1-41.0). The geometric mean concentration of soluble transferrin receptor was lower in the exposed subjects than in the controls (2.2 vs 2.6 mg l(-1); P < 0.001) and the concentration was negatively associated with current exposure to "soluble" Mn in the inhalable aerosol fraction and with current smoking habits. An association was found between the concentration of serum soluble transferrin receptor and the concentration of Mn in whole blood (Pearson's r = 0.48; P < 0.001) in the controls. The results suggest that Mn-exposed workers have higher intracellular iron concentration in the erythrocyte precursors than the controls, resulting in a down-regulation of transferrin receptors on the surface of these cells. The concentrations of Mn in the blood of occupationally non-exposed individuals appear to be influenced by iron status, even at physiological iron levels. PMID- 12884408 TI - Effects of sulfur mustard on transcription in human epidermal keratinocytes: analysis by mRNA differential display. AB - This study examines the transcriptional response of human epidermal keratinocytes (HEK) to sulfur mustard (HD) in order to gain a better understanding of the intracellular events that result in cytotoxicity. Differential display polymerase chain reaction technology was used to examine the relative transcriptional activity of healthy cells to those exposed to subvesicating or vesicating concentrations of HD for 4 h. Approximately 2% of the HEK transcriptome had altered expression. Sixty of the most prominently altered transcripts were characterized. Important upregulated genes include NADH dehydrogenase III, GADD45 and ubiquitin. Key downregulated genes include type I keratin 14, alpha-enolase and caltractin. Many of the identified transcripts protein products presently do not have an assigned function and eleven transcripts were unidentifiable. These transcriptional alterations provide one of the first molecular insights into the intracellular events induced by HD. PMID- 12884409 TI - Resveratrol, a natural aryl hydrocarbon receptor antagonist, protects lung from DNA damage and apoptosis caused by benzo[a]pyrene. AB - Benzo[a]pyrene (BaP) is an agonistic ligand for the aryl hydrocarbon receptor (AhR) and a major environmental carcinogen implicated in the aetiology of lung cancer through the induction of benzo[a]pyrene diol epoxidation (BPDE) and BPDE DNA adducts. Because BaP metabolization requires cytochrome P-450 1A1 (CYP1A1) induction through activation of the AhR, we hypothesized that resveratrol, a natural competitive inhibitor of AhR, could prevent these adverse effects of BaP on the lung. Balb-C mice were injected for 5 weeks with corn oil, BaP (5 mg kg( 1) week(-1)), resveratrol (50 mg kg(-1) week(-1)) or BaP + resveratrol. Immunohistochemistry was performed on lung sections for the determination of CYP1A1 protein, BPDE-DNA adducts and apoptosis. A semi-quantitative immunohistochemistry score (H score) was used for data analysis. Mice exposed to BaP had a significant induction of lung BPDE-DNA adducts when compared with controls (H scores: control, 26, interquartile range 18-33; BaP, 276, interquartile range 269-288; P < 0.01). The BPDE-DNA adduct induction by BaP was abrogated significantly by resveratrol (H score: BaP + resveratrol, 103, interquartile range 96-113). A similar pattern was found by immunohistochemistry for apoptosis (H scores: control, 121, interquartile range 102-137; BaP, 288, interquartile range 282-292, P < 0.05; BaP + resveratrol, 132, interquartile range 121-141, P = NS) and CYP1A1 (H scores: control, 170.3, interquartile range 164-175; BaP, 302.3, interquartile range 291-315, P < 0.05; BaP + resveratrol, 200.7, interquartile range 174-215, P = NS). Western blotting confirmed that resveratrol prevented BaP-induced CYP1A1 expression. This increase in CYP1A1 expression in response to BaP administration most likely causes BaP metabolism, BPDE-DNA adduct formation and subsequent apoptosis. All BaP-induced effects could be prevented by resveratrol, suggesting a possible chemopreventive role for this natural phytoalexin against the development of lung cancer. PMID- 12884410 TI - Neurotoxic character of thimerosal and the allometric extrapolation of adult clearance half-time to infants. AB - The decomposition rate of organomercurials and the potency of the blood-brain barrier increase with the size of the organic radical. Thus methylmercury damages the brain more than thimerosal does, and when intake limits set for methylmercury are applied to thimerosal the safety margin is increased even if the clearances were the same. However, the clearance half-time of ethylmercury in adults is about one-third of the 50 days' clearance half-time of methylmercury given for 60 kg body weight. Moreover, because metabolic rates (e.g. basal metabolism, daily loss of mercury in per cent of body burden) in different weight groups are related to the fractional power of body weight (rule of allometry), mercury clears from the infant body faster than from the adult body. Blood mercury concentrations observed after vaccination showed agreement with allometrically extrapolated concentrations. PMID- 12884411 TI - Testicular and spermatotoxic effects of quinalphos in rats. AB - Testicular and spermatotoxic effects were investigated in rats exposed to technical-grade quinalphos (70%) at dose levels of 0.52 mg kg(-1) (1/50th ld(50)) or 1.04 mg kg(-1) body weight (1/25th ld(50)) for 5 days a week for 60 days. The activities of marker testicular enzymes such as sorbitol dehydrogenase (SDH) and acid phosphatase were significantly decreased but those of lactate dehydrogenase (LDH), gamma-glutamyl transpeptidase (gamma-GT) and beta-glucuronidase were significantly increased in a dose-dependent manner. This particular pattern in the activity of testicular-cell-specific enzymes, a decrease in sperm motility and total epididymal sperm count and an increase in abnormal sperm suggest damage to germ cells and Sertoli cells. The testicular and spermatotoxic effects observed in rats may be due to the pesticide quinalphos or its metabolites. PMID- 12884412 TI - Insights into the mechanism of erythrocyte Na+/K+-ATPase inhibition by nitric oxide and peroxynitrite anion. AB - Evidence shows that Na(+)/K(+)-ATPase from kidney, brain and liver is inhibited by nitric oxide (NO) and peroxynitrite anion (ONO(2) (-)), but the mechanism is unknown. The aim of the present work was to study the inhibitory effect of NO and ONO(2) (-) on erythrocyte Na(+)/K(+)-ATPase. Erythrocyte membranes were isolated from male Wistar rats by hypotonic washing. The membranes (free from haemoglobin) were incubated for Na(+)/K(+)-ATPase activity measurement at various concentrations of ATP in the presence or absence of 400 microM SNAP (an NO donor) or 100 microM SIN-1 (an ONO(2)(-) donor). At these concentrations, SNAP and SIN-1 released about the same amount (100 microM) of NO or ONO(2)(-), respectively, as monitored by measuring NO(2)(-) + NO(3)(-). Both SNAP and SIN-1 decreased V(max) by ca. 75% but they did not decrease the apparent affinity of the Na(+)/K(+) ATPase for the substrate (a decrease of K(m) was even observed after SNAP treatment). The pattern of this inhibition is compatible either with oxidation of SH groups directly involved in ATP binding but in a way that is not surmountable by increasing the substrate concentration ("non-competitive") or with oxidation of SH groups located outside the active site of the enzyme but important for the activity of the enzyme. PMID- 12884413 TI - Harmonization of thresholds for primary skin irritation from results of human repeated insult patch tests and laboratory animal skin irritation tests. AB - Classification of a chemical or chemical product as a primary skin irritant using laboratory animal tests has been defined rigorously and codified in Consumer Products Safety Commission (CPSC) regulations. However, no regulatory agency, including the CPSC, has defined a primary skin irritant threshold for human repeat insult patch tests (RIPTs) that are typically conducted on products, such as cosmetics, anticipated to come into direct contact with humans. Further more, the protocols for animal and human tests are significantly different, as are the schemes for grading responses. Consequently, comparing the results of one type of test with those from the other type has proved to be difficult. In this short communication, we propose a procedure to harmonize the results from these two types of skin irritation tests and suggest that a score of "5" in animal tests, considering 24-h results on unabraded skin, is equivalent to a score of "3" in human RIPTs. PMID- 12884415 TI - Reversal theory states and cigarette availability predict lapses during smoking cessation among adolescents. AB - Although 67% of adolescent smokers say they want to quit, only 7% are able to do so. The purpose of this study was to use metamotivational states as described in reversal theory to predict whether adolescents who are trying to quit smoking will lapse (smoke a cigarette) or will resist smoking in highly tempting situations. Reversal theory holds that individuals switch between telic (serious minded) and paratelic (playful) metamotivational states. Questionnaires and semistructured interviews were administered to 62 adolescents ages 14-19 who had participated in a smoking cessation program. Interviews were coded according to the Metamotivational State Interview and Coding Schedule, and the resulting data were analyzed using logistic regression analysis. Adolescents in the paratelic metamotivational state (OR = 15.34; 95% CI: 5.3, 43.6) or with cigarettes readily available (OR = 4.6; 95% CI: 1.6, 13.2) were more likely to lapse than were adolescents in the telic state or who required effort to obtain cigarettes. The variables telic/paratelic state and cigarette availability accurately predicted the outcome of highly tempting situations 80% of the time. Using reversal theory constructs to develop state-specific strategies shows promise as an additional tool for those in this age group coping with the temptation to smoke. PMID- 12884416 TI - Accuracy in the Outcomes and Assessment Information Set (OASIS): results of a video simulation. AB - There is little information regarding the accuracy of the Outcomes and Assessment Information Set (OASIS), the patient assessment tool mandated for use in Medicare funded home health care. The purposes of this study were to evaluate the accuracy of OASIS completion by home health nurses and rehabilitation therapists, to compare responses of nurses and therapists, and to determine whether dispersion of answers would affect the home health resource group (HHRG) to which patients were assigned for Medicare home health care payments to agencies. Using a video simulation of admission and discharge visits, 436 clinicians from 29 Ohio home health care agencies scored selected OASIS items. Although the majority of the items were rated accurately, discrepancies were found between clinician responses and the "correct" answer on several items. Nurses and therapists provided similar ratings on most items studied, but for most cases in which discrepancies were found, nurses were more likely to agree with the "correct" answer. Discrepancies most often led to patients being assigned to lower-payment HHRGs. Continued monitoring of OASIS data collection accuracy is recommended to maximize the value of the OASIS instrument in home health care research, practice, and policy. PMID- 12884417 TI - Creating avenues for relative empowerment (CARE): a pilot test of an intervention to improve outcomes of hospitalized elders and family caregivers. AB - The purpose of this pilot study was to evaluate the effectiveness of a family caregiver-focused intervention program (CARE) on the outcomes of hospitalized elders and their family caregivers. A randomized clinical trial was conducted with 49 family caregivers of hospitalized elders in a university medical center in upstate New York. Driven by self-regulation and role theories, the two-phase CARE program consisted of: (a). a mutual agreement consisting of family caregiving activities during hospitalization; and (b). audiotaped information regarding emotional responses and possible complications associated with an elderly patient's hospitalization as well as instructions for effectively participating in the elder's hospital care. The comparison program consisted of information about hospital services and policies. CARE elders had fewer incidents of acute confusion reported by family caregivers during hospitalization and fewer depressive symptoms at 2 weeks and 2 months posthospitalization than did the comparison group. CARE family caregivers participated more in the care of their hospitalized elders and had higher scores on role rewards prior to hospital discharge. Findings from this study support the need for further testing of the CARE intervention with family caregivers to determine its effectiveness on outcomes of hospitalized elders and their family caregivers. PMID- 12884418 TI - Mother-infant interactions of medically fragile infants and non-chronically ill premature infants. AB - At 6 months corrected for prematurity, 41 medically fragile prematures, 20 medically fragile full-terms, and 28 prematures without chronic illnesses were observed interacting with their mothers for 1 hr. Mothers of non-chronically ill prematures gestured to and touched their infants less, were uninvolved with them for a longer time, and spent less time interacting and looking at their infants than did mothers of medically fragile infants. Medically fragile full-terms slept more than the non-chronically ill prematures. The non-chronically ill premature group played with objects more and exhibited more locomotion. Thus, the non chronically ill prematures had more mature behaviors but less frequent interactions than did the medically fragile infants. These disparities reflect differences both in the infants' functional maturity and in maternal compensation for infant vulnerability. PMID- 12884419 TI - Using methodological triangulation for cultural verification of commitment to a plan for exercise scale among Korean adults with chronic diseases. AB - The purpose of this study was twofold: to use methodological triangulation to increase the cultural appropriateness of the Planning for Exercise Scale (Pender, 1996), which measures commitment to a plan of action, and to psychometrically evaluate the revised scale in a selected population of 230 Korean adults experiencing chronic diseases. First, through a qualitative approach, the cultural properties of the concept were identified in order to revise Pender's Planning for Exercise Scale. The revised scale consisted of 20 items: 13 items emerged from the qualitative procedure and seven items were retained from the original scale. Through a quantitative approach, the revised scale was analyzed psychometrically. Results showed the scale had sufficient reliability and validity to warrant its use by researchers or practitioners evaluating the level of commitment to a plan for exercise among Korean adults. The revised instrument is appropriate for use in future intervention studies designed to improve the performance of exercise among Koreans who are chronically ill. PMID- 12884420 TI - Research methods: managing primary study quality in meta-analyses. AB - Meta-analyses synthesize multiple primary studies and identify patterns of relationships. Differences in primary study methodological quality must be addressed for meta-analysis to produce meaningful results. No single standard exists for addressing these quality variations. Quality measurement scales are fraught with development and application problems. Several strategies have been proposed to address quality. Researchers can set minimum levels for inclusion or require that certain quality attributes be present. An inclusive method is to weight effect sizes by quality scores. This allows the inclusion of diverse studies but relies on questionable quality measures. By considering quality an empirical question, meta-analysts can examine associations between quality and effect sizes and thus preserve the purpose of meta-analysis to systematically examine data. Researchers increasingly are combining strategies to overcome the limitations of using a single approach. Future work to develop valid measures of primary study quality dimensions will improve the ability of meta-analysis to inform research and nursing practice. PMID- 12884421 TI - Living with achondroplasia in an average-sized world: an assessment of quality of life. AB - Mutations in the gene encoding fibroblast growth factor receptor 3 cause achondroplasia, the most common form of inherited skeletal dysplasia. Although there are more than 10,000 individuals with achondroplasia living in the United States, there has been little study of their quality of life (QOL). For this study, surveys were collected from 189 individuals affected with achondroplasia (ACH) and 136 unaffected first-degree relatives (FDRs) of affected individuals. Individuals affected with achondroplasia had lower annual income, less education, and were less likely to be married than FDRs. They also differed significantly in their perceptions of achondroplasia, with FDRs believing that achondroplasia is a more serious condition. Total QOL indices and indices in each of four QOL subdomains were significantly lower in affected individuals than in relatives. When controlling for demographic characteristics and affected status, having lower self-esteem scores and perceiving achondroplasia as more serious were the independent factors most highly associated with lower QOL. A qualitative analysis of open responses to questions about the advantages and disadvantages of achondroplasia revealed that individuals were as likely to cite disadvantages relating to social barriers as they were to cite those relating to health and functioning. We interpret the low QOL scores to reflect the social challenges that individuals with achondroplasia regularly experience in the average-sized world. Genetics professionals should consider sources of lower QOL for affected individuals in their counseling sessions to acknowledge the relative importance of non-medical contributions. PMID- 12884422 TI - Case-control study of cleft lip or palate after maternal use of topical corticosteroids during pregnancy. AB - A case-control survey of 48 children with nonsyndromic cleft lip or palate showed a significant increase in prevalence of maternal use of topical corticosteroid preparations in the first trimester of pregnancy, compared to 58 controls born in the same hospital; the odds ratio was 13.154, 95% confidence interval 1.67-586, P = 0.0049 on Fisher's exact two-tail test. The results were statistically significant although the wide confidence interval reflected the small sample size. Although older epidemiological studies have not detected any association between systemic corticosteroid treatment and the combined incidence of all congenital malformations, experimental studies over several decades have shown that maternal corticosteroid exposure in several species of animals is specifically associated with oral clefts. This association has been confirmed by more specific case-control surveys where the cases were children with cleft lip or palate and the exposure was maternal systemic corticosteroid treatment in the first trimester. Only one previous survey also analyzed topical corticosteroids, and it demonstrated an increased odds ratio for cleft lip or palate. A national survey of children with cleft palate will be necessary to evaluate the results of this pilot study. PMID- 12884423 TI - Prenatal detection of mosaic trisomy 1q due to an unbalanced translocation in one fetus of a twin pregnancy following in vitro fertilization: a postzygotic error. AB - Complete or mosaic trisomy for all of chromosome 1q has been seen rarely in a recognized pregnancy. A patient presented with twins following in vitro fertilization (IVF). Ultrasound showed twin A to have a diaphragmatic hernia, thick nuchal fold, and subtle intracranial abnormalities. Twin B appeared normal and a thick dividing membrane was seen. Amniocentesis of twin A showed a male karyotype with mosaic trisomy 1q in 57% of cells resulting from a translocation between chromosomes Yq12 and 1q12. Parental karyotypes were normal. The twins were delivered at 33 weeks. Twin A died at 1 hr of life. Autopsy confirmed the left diaphragmatic hernia and hypoplastic lungs. Autopsy also revealed a partial cleft palate, syndactyly of the second and third toes bilaterally, external deviation of the left 5th toe, and contractures of the index fingers bilaterally. A recent report documented formation of a chimera resulting from embryo amalgamation after IVF. Given the rarity of the cytogenetic findings in our case, we sought to determine if the mosaicism was a result of chimera formation related to the IVF. Thirteen polymorphic loci throughout the genome, in addition to four on 1q and four on 1p, were amplified by PCR. Only two alleles were observed at each of these loci in twin A, one paternal and the other maternal. We present further clinical findings of this case with a rare cytogenetic abnormality that appears to have originated from a postzygotic mitotic error and not embryo amalgamation. PMID- 12884424 TI - Screening of patients with craniosynostosis: molecular strategy. AB - Craniosynostosis is the premature fusion of calvarial bones leading to an abnormal head shape. The craniosynostosis syndromes are clinically heterogeneous with overlapping features, which make an accurate diagnosis difficult at times. Although the clarification of a genetic lesion does not have a direct impact on patient management in many cases, there is a significant benefit in providing accurate prenatal diagnosis. Genetic counsellors are also able to offer better risk estimates of recurrences to non-manifesting carriers and their extended family members and for affected patients of reproductive age. Advances in gene discovery have shown that craniosynostosis syndromes delineated on clinical bases, with the possible exception of Apert syndrome, are genetically heterogeneous, and mutations have been found in fibroblast growth factor receptors (FGFR) 1, 2, 3 and TWIST. We surveyed 99 craniosynostosis patients at the molecular level and found mutations in 50 of them. Six novel point mutations were identified: three in FGFR2 and three in TWIST. Two Saethre-Chotzen patients with TWIST microdeletions at 7p21 were also found. The other mutations identified have been previously reported. In studying these 99 patients, we developed a diagnostic strategy for craniosynostosis testing, where sequential analysis of recurrent mutations was followed by selective sequencing. This algorithm makes testing of craniosynostosis disorders more efficient and cost-effective. PMID- 12884425 TI - Maternal inheritance in cyclic vomiting syndrome with neuromuscular disease. AB - Cyclic vomiting syndrome (CVS), characterized by severe discrete episodes of nausea, vomiting, and lethargy, is a predominately childhood condition associated with migraine and dysautonomic features. Disease-associated mitochondrial DNA (mtDNA) sequence variants are suggested by a strong maternal bias in the inheritance of migraine, and the recent findings of mtDNA variants in a few children with CVS and additional neuromuscular disease manifestations ("CVS+"). A clinical interview using a questionnaire was administered (generally) to one parent of 62 children with CVS+. Non-senile disease manifestations, including migraine, myopathy, seizures, and dysautonomia-like symptoms, were far more common in matrilineal versus non-matrilineal relatives, including being present in 75% of the mothers versus in only 11% of the fathers (P < 0.001). Overall, maternal inheritance is suggested in 86% of the families (in 65% strongly so). Disease manifestations in subjects and their affected matrilineal relatives are predominately intermittent and consistent with dysautonomia, including increased vital sign fluctuations. Body fluid metabolites and muscle biopsy findings are consistent with mitochondrial dysfunction in most cases tested. We conclude that mtDNA sequence variants are at least risk factors in the development of disease in most children at this "severe" end of the CVS spectrum, likely involving a maternally inherited propensity towards dysautonomia. PMID- 12884426 TI - Characterization of terminal chromosome anomalies using multisubtelomere FISH. AB - Telomeric repeat sequences (TTAGGG) are known to cap the termini of every human chromosome. Proximal to these repeat sequences are chromosome-specific repeat sequences, which in turn are distal to gene-rich regions. Submicroscopic, subtle, or cryptic abnormalities in these regions can now be investigated using commercial probe sets for all of the chromosome-specific subtelomeric regions of the human genome. Using this technology, previously unidentified genomic imbalance has been found in a proportion of patients with idiopathic developmental delay and learning difficulties. We have used these probe sets to investigate cases with apparently terminal anomalies detected on G-banded chromosome analysis. As a result of such investigations, we have found that 3 (19%) of 16 apparently terminal deletion cases were the result of more complex rearrangements involving other chromosome subtelomeres. The remaining 13 cases contained no chromosome-specific subtelomere repeats on the deleted arm, but in all 16 cases, the TTAGGG telomere repeat cap was present. A further case was investigated where extra material was found in the terminal region of the chromosome 12 short arm, found to represent a complex inversion/duplication/deletion rearrangement. Investigation of all cases with terminal anomalies, including apparently terminal deletions, is likely to uncover further cases involving complex rearrangements and should lead to a greater understanding of the mechanisms by which these abnormalities arise. PMID- 12884427 TI - Familial multiple epiphyseal dysplasia due to a matrilin-3 mutation: further delineation of the phenotype including 40 years follow-up. AB - In this study, we followed-up the family with bilateral hereditary micro epiphyseal dysplasia (BHMED) originally described by Elsbach [1959: J Bone Joint Surg [Br] 41-B:514-523]. Clinical re-examination of all available family members resulted in further delineation of the clinical and radiological phenotype, which is distinct from common multiple epiphyseal dysplasia (MED). Linkage analysis excluded EDM1, EDM2, and EDM3 as candidate genes. Linkage and mutation analysis of matrilin-3 (MATN-3) revealed a new pathogenic mutation confirming that BHMED is indeed a distinct disease entity among MED and MED-like disorders. PMID- 12884428 TI - Spondyloepiphyseal dysplasia Maroteaux type: report of three patients from two families and exclusion of type II collagen defects. AB - Spondyloepiphyseal dysplasia (SED) Maroteaux type is an autosomal dominant skeletal dysplasia, characterized by spondylar dysplasia, mild epiphyseal dysplasia of the large joints, and type E-like brachydactyly. These manifestations overlap with those of spondyloperipheral dysplasia (SPD), in which a sporadic case with a mutation of COL2A1 has been reported. We report on three patients (an affected woman and her son and a sporadic case of an affected man) with SED Maroteaux type. The affected adults were severely short along with stubby hands and feet, and one developed myelopathy as a result of thoracolumbar gibbus. The affected child was mildly short at birth, and developed brachydactyly in early childhood. The radiological hallmarks of these patients included severe platyspondyly with square-shaped vertebral bodies, iliac hypoplasia, epiphyseal hypoplasia of the large joints, and strikingly short metacarpals and phalanges. These radiological findings appeared already apparent in early childhood. SED Maroteaux type was radiologically discriminative from SPD. Brachydactyly was much severe in the former than in the latter, and spondylar dysplasia manifestation was different between both disorders. Mutation screen by polymerase chain reaction-direct sequencing for all exons and their flanking regions of COL2A1 did not reveal any mutations in the three patients. PMID- 12884429 TI - Sirenomelia sequence according to the distance between the first sacral vertebra and the ilia. AB - The development of the iliac bones and lower limbs are parallel processes depending on the normal ontogeny of the caudal blastema [O'Rahilly and Muller, 1989; Opitz, 1993; Opitz et al., 2000]. We hypothesized that the pathogenetic mechanisms leading to fused lower limbs would in parallel displace the ilia caudally and medially and that the degree of this displacement might correlate with the severity of the iliac and lower limb defects. Thus the purpose of the study was to test this hypothesis in a sample of 12 sirenomelic fetuses. The fetuses GA 16-39 weeks, spontaneously or therapeutically aborted, were radiographed in the frontal projection as part of a requested autopsy. From each radiograph, a line was drawn connecting the most cranial part or the two ilia. After that the distance was measured vertically between this line and the most cranial part of the first sacral vertebral body (iliac-sacral distance (ISD)). A second distance was measured horizontally between the most lateral part of the two iliae (bi-iliac distance (BD)). As a result, ISD correlates with the iliac/femur phenotype. Separate ilia and femora occur only in cases with normal ISD. Fused ilia or femora or both are seen only in fetuses with mildly increased ISD, whereas a single iliac bone and femur occur only in cases with greatly increased ISD. The increase of the ISD does not correlate with the severity of more distal limb involvement. There was a correlation between the ISD and the BD values; the higher the ISD, the shorter the BD. Based on these findings, we propose an extended classification of Sirenomelia to be tested by other researchers. PMID- 12884430 TI - X-linked mild non-syndromic mental retardation with neuropsychiatric problems and the missense mutation A365E in PAK3. AB - We describe a family of 19 males in five generations with mild to borderline non syndromic X-linked mental retardation (MRX). There were no clinical manifestations in the affected males other than mental impairment and relatively long ears, with neuropsychiatric problems in some cases. Linkage analysis carried out on part of the pedigree using 34 markers spanning the X chromosome localized the gene between DXS454 and DXS1001 in Xq23. The maximum two-point lod score was 3.21 at DXS1059. PAK3 is a known MRX gene mapping to the same region. The affected males and obligate carrier females were found to have a missense mutation c.1094C > A in exon 10 causing an A365E substitution in a highly conserved region of the protein. The C to A base change abolishes a PvuII restriction enzyme site providing the basis for a simple test, if required, for carrier detection and prenatal diagnosis in the extended family. PMID- 12884431 TI - Splenogonadal fusion-limb defect "syndrome" and associated malformations. AB - Two types of splenogonadal fusion (SGF) have been proposed. In the continuous type the spleen is connected to the gonad, and often it is associated with limb defects and other anomalies such as micrognathia, microglossia, anal atresia, and hypoplastic lungs. Associated abnormalities are lower in the discontinuous type of SGF, where there is fusion between the gonad and accessory splenic tissue, without connection with the normal spleen. The cause of SGF and SGF with limb defects (SGFLD) is unknown. On the basis of associated orofacial limb abnormalities, some authors have suggested that SGFLD may be related to the Hanhart complex. Others have noted the overlap between SGFLD and femoral-facial syndrome (FFS). We consider SGFLD a developmental field defect that originates during blastogenesis. Although the cause is unknown, the earlier its action, the more severe the involvement resulting in SGFLD; later action may result only in SGF. PMID- 12884432 TI - D-2-hydroxyglutaric aciduria: a case with an intermediate phenotype and prenatal diagnosis of two affected fetuses. AB - D-2-hydroxyglutaric aciduria (D2HGA) is a rare autosomal recessive disorder with variable clinical expression. The biochemical defect is unknown at present. Previously reported cases have either followed a severe clinical course characterized by neonatal epileptic encephalopathy, cortical blindness, and profound developmental delay, or a mild course characterized by mild developmental delay, manageable epilepsy, and mild hypotonia. To date there has been a clear distinction between these two groups. We report the second case of a child with D2HGA who has followed an intermediate course. She presented in infancy with hypotonia, manageable epilepsy and developed moderate to severe developmental delay, and cortical visual impairment. The proposita had a coarse facial appearance, flat face, broad nasal bridge, up-turned nose, and simple, anteverted ears. These facial anomalies have been noted in other children with D2HGA and this case strengthens the proposed association between this facial phenotype and D2HGA. We also report the third and fourth instances of prenatal diagnosis for D2HGA. At each prenatal diagnosis, an affected fetus was diagnosed on the basis of markedly increased levels of D-2-hydroxyglutaric acid in amniotic fluid. PMID- 12884433 TI - Abnormalities of the umbilico-portal venous system in Down syndrome: a report of two new patients. AB - Congenital anomalies of the umbilical and portal venous system are rare vascular malformations which are often associated with anomalies of the heart and gastrointestinal tract. Association with chromosomal disorders has been sporadically reported. We now report on two patients with trisomy 21 and congenital anomalies of the umbilico-portal system. A male fetus showed absence of the intrahepatic portal vein (PV) and ductus venosus with a direct communication between portal sinus and inferior vena cava exhibiting an umbilicosystemic total shunt during the fetal life and a portosystemic total shunt after birth. A female infant showed absence of the intrahepatic PV and a total portocaval shunt. Both patients also had heart defects. As previously documented in other reports, our cases demonstrated that this association may be causally-related to the chromosomal aberration. In addition, the umbilico-portal venous system abnormalities seems to be the most frequent congenital vascular malformation in Down syndrome. A presumptive pathogenetic mechanism could be a trisomy 21-related altered angiogenesis of the vitelloumbilical plexus. PMID- 12884434 TI - Autistic disorder and chromosomal mosaicism 46,XY[123]/46,XY,del(20)(pter --> p12.2)[10]. AB - We report on a 3-year-old boy with a moderate to severe mental retardation, autistic behavior patterns, and myoclonic epilepsy of early childhood. The cytogenetic analysis of blood lymphocytes revealed a deletion of chromosome 20pter --> p12.2 occurring as mosaicism in 8% of the analyzed metaphases: 46,XY[123]/46,XY,del(20)(pter --> p12.2)[10]. The deletion was confirmed by the recently developed multicolor banding approach and additionally by region specific fluorescence in situ hybridization (FISH) probes. To the best of our knowledge, this is the first report on a patient with autistic behavior with terminal 20p deletion mosaicism reported up to present. PMID- 12884435 TI - Toriello-Carey syndrome associated with respiratory failure and non-mechanical ileus. AB - Toriello-Carey syndrome comprises agenesis of the corpus callosum, telecanthus, small palpebral fissures, Pierre Robin sequence, abnormal ears, and cardiac defects. We report a boy who has some additional findings, including a severe respiratory failure and intestinal dysmotility. The boy died of these two disorders at age 13 months. Histological examination revealed pulmonary immaturity and a defect of smooth muscle cells in the longitudinal muscle coat of the intestinal musculature, both of which might explain some aspects of the pathophysiology of the patient. PMID- 12884436 TI - Craniosynostosis and congenital heart anomalies associated with a maternal deletion of 15q15-22.1. AB - We report an infant with multiple congenital anomalies, including craniosynostosis, tetralogy of Fallot variant, and limb anomalies associated with a maternal deletion of 15q15-22.1. Only two other patients have been reported with a similar deletion, but the deletion was paternal in both cases. We review our patient's findings and compare them to previously reported individuals with similar 15q abnormalities. Our patient allows an expansion of phenotype associated with mid-15q deletions to include severe craniosynostosis, congenital heart disease, and limb anomalies. This will assist in prenatal counseling and predicting postnatal outcome for other affected individuals. The specific breakpoints in our patient and the other patients with similar deletions may also assist in determining a critical region for suture formation. PMID- 12884437 TI - Antenatal onset of cortical hyperostosis (Caffey disease): case report and review. AB - We report on a fetus of 27 weeks of gestation whose clinical, radiological, and histopathological findings are compatible with the prenatal form of Caffey disease (cortical hyperostosis). Prenatal ultrasound examination showed polyhydramnios and markedly short and angulated long bones, which had led to the incorrect diagnosis of lethal osteogenesis imperfecta. We found 43 published descriptions on the antenatal onset of cortical hyperostosis symptomatic in utero or at birth. Two groups of prenatal cortical hyperostosis (PCH) can be distinguished: (1) severe (25 reports and the fetus presented here), with onset before 35 weeks of gestation and generally associated with polyhydramnios, lung disease, prematurity, and high lethality; (2) mild (18 reports), with onset after 35 weeks of gestation and without complications. Autosomal recessive inheritance has been suggested for the prenatal form of cortical hyperostosis. However, the available evidence suggests that both dominant and recessive inheritance is possible. Moreover, dominant inheritance seems to be more common in the mild prenatal form of cortical hyperostosis. PMID- 12884438 TI - Severely delayed epiphyseal ossification dysplasia with normal stature. AB - We report on a pair of dizygotic twins born to consanguineous parents, showing an extreme delay in skeletal maturation (roughly corresponding to the time of birth at the age of 4 10/12 years), mild metaphyseal irregularities, and normal to large stature (+ 1 to + 2 SD) with normal growth pattern, long fingers, generalized small joint hyperlaxity with flat feet, genua valga, and low normal intellectual development. This disorder was not associated with abnormal endocrine function or abnormal Ca/P metabolism. This epiphyseal disorder is compatible with autosomal recessive inheritance. PMID- 12884439 TI - Functional disomy for Xq22-q23 in a girl with complex rearrangements of chromosomes 3 and X. AB - A 5-year-old girl with developmental and growth retardation is reported with complex chromosome rearrangements consisting of a partial Xq deletion and an abnormal chromosome 3 with multiple breakpoints. GTG-banding, and multiplex and conventional FISH studies showed that a 6.6-Mb Xq22-q23 segment was inserted into 3q, in addition to three intrachromosomal insertions in chromosome 3. Her karyotype was thus interpreted as 46,X,der(X)(Xpter-->Xq22::Xq23- >Xqter),der(3)(3pter-->3p26::3p12-->3q25.3::3p12-->3p26::Xq22-->Xq23::3q25.3- >3qter). Replication R-banding study showed that the der(X) was inactivated in all blood lymphocytes analyzed. Methylation-specific PCR at the androgen receptor gene (HUMARA) locus at Xq11-q12 showed a skewed inactivation pattern with the active/inactive X chromosome ratio of 92/8. These data indicated the presence, in the majority of cells, of a functioning Xq22-q23 segment in both the normal X and the der(3) chromosomes. Her growth retardation, developmental delay, and other minor anomalies were most likely caused by dosage effects of the genes in the functionally disomic Xq22-q23 region. Despite the presence of two active copies of the proteolipid protein 1 gene (PLP1), she did not show the symptoms of Pelizaeus-Merzbacher disease, a subset of which has been known to be caused by the duplication of PLP1. PMID- 12884440 TI - Sliding type hernia and ectopic pancreatic tissue in the stomach with renal agenesis and ear abnormalities: branchio-oto-renal syndrome or hemifacial microsomia with additional findings. PMID- 12884441 TI - On macrocephaly, epilepsy, autism, specific facial features, and mental retardation. PMID- 12884442 TI - Do genetic polymorphisms of serotonin (5-HT) neurotransmission influence function in humans? PMID- 12884443 TI - Conjoined twins: morphogenesis of the heart and a review. AB - Five cases of conjoined twins have been studied. These included three thoracopagus twins, one monocephalus diprosopus (prosop = face), and one dicephalus dipus dibrachus. The thoracopagus twins were conjoined only from the upper thorax to the umbilicus with a normal foregut. These three cases shared a single complex multiventricular heart, one with a four chambered heart with one atrium and one ventricle belonging to each twin with complex venous and arterial connection; two had a seven chambered heart with four atria and three ventricles. The mono-cephalus diprosopus twins had a single heart with tetralogy of Fallot. The dicephalus twins had two separate axial skeletons to the sacrum, two separate hearts were connected between the right atria with a shared inferior vena cava. Thoracopagus twinning is associated with complex cardiac malformations. The cardiac anlagen in cephalopagus or diprosopus are diverted and divided along with the entire rostral end of the embryonic disc and result in two relatively normal shared hearts. However, in thoracopagus twins the single heart is multiventricular and suggests very early union with fusion of the cardiac anlagen before significant differentiation. Cardiac morphogenesis in conjoined twins therefore appears to depend on the site of the conjoined fusion and the temporal and spatial influence that determines morphogenesis as well as abnormally oriented embryonic axes. PMID- 12884444 TI - Developmental anomalies of the scapula-the "omo"st forgotten bone. AB - Congenital malformations of the scapula, ranging from complete absence, to abnormal shape and position (Sprengel anomaly) are encountered, not infrequently, in genetic practice. Despite this, little is known of the embryologic origin of the scapula and the relationship of the embryology to the observed birth defects. Standard embryology texts, when discussing the subject at all, generally consider the scapula as part of the upper limb. The pattern of associated birth defects suggests that this is at least an oversimplification and may be inaccurate. Sprengel anomaly is the most frequently encountered malformation of the scapula. It can be seen in isolation, but is often seen in association with other defects that include; scoliosis, hemivertebrae, segmentation abnormalities of vertebrae and ribs (including Klippel-Feil sequence), spina bifida, clavicular abnormalities, renal abnormalities and hypoplasia of the muscles of the neck and shoulder. The ipsilateral limb is usually normal. An unappreciated association between Sprengel anomaly and diastematomyelia of the lumbar spine also raises questions about the embryologic origin of the scapula. 25-50% of Sprengel anomaly patients have an associated omovertebral band or bone that arises from the posterior process of a vertebral body and attaches to the superior angle/medial portion of the scapula. This is felt to be of scapular origin, but the report of at least one patient with an omovertebral bone, not associated with a Sprengel anomaly questions this assumption. Scapuloiliac dysostosis (Kosenow syndrome), a rare skeletal dysplasia, is associated with marked hypoplasia of the scapulae, clavicles and pelvis. Associated anomalies include eye anomalies, rib anomalies and spina bifida. The limbs are normal. Knockout of the Emx2 gene in mice yields a similar skeletal phenotype. Mutations in EMX2 in humans are associated with schizencephaly, not skeletal anomalies. Data on gene expression in the scapula will be reviewed. Based on this information, the author proposes that the scapula arises from 2 or more distinct embryologic anlage under different genetic control than the upper limb. PMID- 12884445 TI - Preempting genetic discrimination and assaults on privacy: report of a symposium. AB - At a symposium in June, 2002, biomedical researchers, clinicians, legal experts, policymakers, and representatives of the insurance industry and the advocacy community gathered to address issues of genetic privacy and discrimination; and to identify research, legal, and policy gaps needing to be filled. They concluded that over the next decade, as more genetic information becomes available and the public becomes more aware of individual risks, concerns about privacy and discrimination will become increasingly important. Documented cases of genetic discrimination are rare and largely anecdotal, yet individuals with genetic conditions harbor significant fears about discrimination. Current laws enacted to protect individuals from workplace and insurance discrimination offer some measure of protection, but leave many unfilled gaps. Moreover, the use of genetic information in potentially discriminatory ways is not limited to employment and insurability. Existing laws do little to protect people seeking life, disability, or long-term care insurance. And the courts have used genetic information in a wide variety of cases including paternity, criminal, and tort (personal injury) cases. Genetic information that might jeopardize an individual's right to privacy may also be obtained in the course of research studies, including through the collection of DNA and tissue samples. The insurance industry, State and Federal agencies, and the advocacy community are all making efforts to address some of these gaps through legislation and education of clinicians, the public, and policy makers. PMID- 12884446 TI - Genetic Drift. Prenatal diagnosis is for the DR, not just for the OR. PMID- 12884447 TI - Estimate of the prevalence of chromosome 15q11-q13 duplications. PMID- 12884448 TI - Associative memory design using overlapping decomposition and generalized brain state-in-a-box neural networks. AB - This paper is concerned with large scale associative memory design. A serious problem with neural associative memories is the quadratic growth of the number of interconnections with the problem size. An overlapping decomposition algorithm is proposed to attack this problem. Specifically, a pattern to be processed is decomposed into overlapping sub-patterns. Then, neural sub-networks are constructed that process the sub-patterns. An error correction algorithm operates on the outputs of each sub-network in order to correct the mismatches between sub patterns that are obtained from the independent recall processes of individual sub-networks. The performance of the proposed large scale associative memory is illustrated using two-dimensional images. It is shown that the proposed method reduces the computing cost of the design of the associative memories compared with non-interconnected associative memories. PMID- 12884449 TI - Singular value decomposition learning on double Stiefel manifold. AB - The aim of this paper is to present a unifying view of four SVD-neural computation techniques found in the scientific literature and to present some theoretical results on their behavior. The considered SVD neural algorithms are shown to arise as Riemannian-gradient flows on double Stiefel manifold and their geometric and dynamical properties are investigated with the help of differential geometry. PMID- 12884450 TI - Global and robust stability of interval Hopfield neural networks with time varying delays. AB - In this paper, we investigate the problem of global and robust stability of a class of interval Hopfield neural networks that have time-varying delays. Some criteria for the global and robust stability of such networks are derived, by means of constructing suitable Lyapunov functionals for the networks. As a by product, for the conventional Hopfield neural networks with time-varying delays, we also obtain some new criteria for their global and asymptotic stability. PMID- 12884451 TI - Cost component analysis. AB - In optimizations the dimension of the problem may severely, sometimes exponentially increase optimization time. Parametric function approximatiors (FAPPs) have been suggested to overcome this problem. Here, a novel FAPP, cost component analysis (CCA) is described. In CCA, the search space is resampled according to the Boltzmann distribution generated by the energy landscape. That is, CCA converts the optimization problem to density estimation. Structure of the induced density is searched by independent component analysis (ICA). The advantage of CCA is that each independent ICA component can be optimized separately. In turn, (i) CCA intends to partition the original problem into subproblems and (ii) separating (partitioning) the original optimization problem into subproblems may serve interpretation. Most importantly, (iii) CCA may give rise to high gains in optimization time. Numerical simulations illustrate the working of the algorithm. PMID- 12884452 TI - Global convergence of delayed neural network systems. AB - In this paper, without assuming the boundedness, strict monotonicity and differentiability of the activation functions, we utilize a new Lyapunov function to analyze the global convergence of a class of neural networks models with time delays. A new sufficient condition guaranteeing the existence, uniqueness and global exponential stability of the equilibrium point is derived. This stability criterion imposes constraints on the feedback matrices independently of the delay parameters. The result is compared with some previous works. Furthermore, the condition may be less restrictive in the case that the activation functions are hyperbolic tangent. PMID- 12884453 TI - A hybrid neural network of addressable and content-addressable memory. AB - We investigate the memory structure and retrieval of the brain and propose a hybrid neural network of addressable and content-addressable memory which is a special database model and can memorize and retrieve any piece of information (a binary pattern) both addressably and content-addressably. The architecture of this hybrid neural network is hierarchical and takes the form of a tree of slabs which consist of binary neurons with the same array. Simplex memory neural networks are considered as the slabs of basic memory units, being distributed on the terminal vertexes of the tree. It is shown by theoretical analysis that the hybrid neural network is able to be constructed with Hebbian and competitive learning rules, and some other important characteristics of its learning and memory behavior are also consistent with those of the brain. Moreover, we demonstrate the hybrid neural network on a set of ten binary numeral patters PMID- 12884454 TI - Diabetes diet war. PMID- 12884455 TI - Do you feel like bug bait? PMID- 12884456 TI - In no mood for therapy. PMID- 12884457 TI - Our mothers' bones. PMID- 12884458 TI - Adalimumab (HUMIRA): a review. AB - Adalimumab (HUMIRA, Abbott Laboratories) is a new fully human TNF-alpha monoclonal antibody recently approved for the treatment of rheumatoid arthritis and undergoing trials for use in treating other conditions, including psoriasis and psoriatic arthritis. This article reviews its mechanisms of action, clinical trial results, and related discussion. PMID- 12884459 TI - Linezolid and quinupristin/dalfopristin: novel antibiotics for gram-positive infections of the skin. AB - With the continuing development of clinical drug resistance among bacteria, the need for new, effective agents to treat multi-drug-resistant Gram-positive infections remains important. With treatment options limited, it has become critical to identify antibiotics with novel mechanisms of activity. Several new drugs have emerged as possible therapeutic alternatives. This review focuses on agents newly introduced and FDA-approved for the treatment of skin and skin structure infections: linezolid and quinupristin/dalfopristin. PMID- 12884460 TI - The role of apoptosis in human epidermal keratinocytes. AB - The mechanisms of apoptosis have been extensively studied in certain cell types such as lymphocytes. However, while it is known that apoptosis is an intrinsic part of the turnover of normal human keratinocytes, relatively little is known about how this cell population utilizes programmed cell death to maintain cutaneous homeostasis. The apoptotic pathways thought to be employed by epidermal keratinocytes in the various cell layers are reviewed, with special emphasis on the protective mechanisms such as proto-oncogenes bcl-2 and Bcl-XL, growth factors, and the NFkB pathway in protecting keratinocytes from premature apoptosis during the process of upward migration and differentiation. The similarities and distinctions between terminal differentiation and apoptosis in keratinocytes are discussed. Both the passive and active apoptosis, including the TNF alpha and Fas-mediated pathways are highlighted, with regards to utilization in normal human epidermal turnover. A firm understanding of the mechanisms of apoptosis in normal human epidermis may allow dermatologists to further appreciate the aberrancies of this process in psoriatic epidermis, and impact on future targets by which to treat hyperproliferative disease. PMID- 12884461 TI - A single-center study of aminolevulinic acid and 417 NM photodynamic therapy in the treatment of moderate to severe acne vulgaris. AB - This article describes a single-center trial involving twenty-two patients with acne vulgaris. Blu-U light was used in all patients, while half were pre-treated with aminolevulinic acid. Preliminary results show promise for this treatment in mild to moderate inflammatory acne vulgaris. PMID- 12884462 TI - Current trend in ivermectin usage for scabies. AB - Ivermectin, an anti-parasitic agent widely used for onchocerciasis in humans, is emerging as an oral antiscabietic that is as safe and effective as the topical antiscabietics. In the recent reports, all groups of population responded to ivermectin in the treatment of scabies, including immunocompetent, immunocompromised, and other high-risk populations such as individuals with Down's syndrome. This report reviews the efficacy, the mechanism of action, and the safety profile of ivermectin in the treatment of scabies, particularly its utility in crusted scabies and outbreaks of scabies in institutional settings. PMID- 12884463 TI - Vitiligo therapy: where are we now? AB - Vitiligo is a disfiguring skin disease. Many insights into its pathogenesis have been identified in recent years; however, treatment remains a challenge. In this article, the various treatment options for the treatment of vitiligo are outlined and newer treatment options are discussed. PMID- 12884464 TI - Systemic contact dermatitis to doxepin. AB - Although allergic contact dermatitis to topical preparations of doxepin has been published, systemic contact dermatitis from oral doxepin is more of a theoretical consideration and is rarely reported. We report a case of a patient with contact allergy to doxepin hydrochloride 5% cream who developed a systemic contact dermatitis to oral doxepin. PMID- 12884465 TI - Erythema elevatum diutinum in an HIV-positive patient. AB - A 53-year old man with HIV infection and hepatitis C infection presented with multiple, firm nodules on the hands and ankles. Erythema elevatum diutinum is a rare, chronic cutaneous vasculitis that may be associated with hematologic, autoimmune, and infectious diseases, which include HIV infection. First-line therapy includes dapsone, as well as treatment of any underlying cause or infection. PMID- 12884466 TI - The use of infliximab in cutaneous sarcoidosis. AB - Infliximab, a chimeric (humanized mouse) monoclonal antibody which specifically inhibits TNF-alpha, has recently shown efficacy in the treatment of refractory sarcoidosis. The manifestations of sarcoidosis are diverse; however, the long term effects on the lungs are often the most life-threatening. We describe a patient with sarcoidosis who experienced the complete resolution of her cutaneous disease and stabilization of her pulmonary symptoms after initiation of infliximab. PMID- 12884467 TI - Case and discussion of Lyme disease. AB - Lyme disease is a multisystem disease with prominent neurological involvement, primarily affecting the nervous system, skin, joints, and heart. We report the case of a patent with Lyme Disease and discuss the background as treatment of this condition. PMID- 12884468 TI - Annular erythema responding to tacrolimus ointment. AB - Annular erythema is an unusual, often idiopathic disorder that tends to respond poorly to topical therapy. Two patients with idiopathic, topical corticosteroid resistant annular erythema showed prompt clearing of lesions treated with 0.1% tacrolimus ointment and persistence of untreated ones which themselves responded to subsequent treatment. These two cases demonstrate a clear-cut therapeutic response of chronic, topical corticosteroid-resistant annular erythema to topical tacrolimus ointment 0.1% BID. Additional experience with tacrolimus ointment, hopefully in controlled circumstances, should clarify its potential value in treating annular erythema. PMID- 12884469 TI - Cardiac catheterization-induced acute radiation dermatitis presenting as a fixed drug eruption. AB - Acute radiation dermatitis is an uncommon complication of fluoroscopic procedures. Cases secondary to cardiac catheterization are infrequently reported in the literature. The risk of developing this reaction is directly related to the skin dose of radiation, which is affected by the type of procedure, the technique used to achieve appropriate penetration, the duration of exposure, and the patient's body habitus. We report a case of acute radiation dermatitis following prolonged coronary angiography initially presenting as a fixed drug eruption. PMID- 12884470 TI - New and emerging treatments for nonmelanomas and actinic keratoses. PMID- 12884471 TI - Systemic and topical drugs for aging skin. AB - The rejuvenation of aging skin is a common desire for our patients, and several options are available. Although there are some systemic methods, the most commonly used treatments for rejuvenation of the skin are applied topically. The most frequently used topical drugs include retinoids, alpha hydroxy acids (AHAs), vitamin C, beta hydroxy acids, anti-oxidants, and tocopherol. Combination therapy is frequently used; particularly common is the combination of retinoids and AHAs. Systemic therapies available include oral retinoids and vitamin C. Other available therapies such as chemical peels, face-lifts, collagen, and botulinum toxin injections are not discussed in this article. PMID- 12884472 TI - [Visibility of scientific journals and initiatives to improve the diffusion of spanish publications]. AB - The conceptual change which has been produced in the transmission of technology knowledge among the scientific community is examined. The international and national systems to organize, store and retrieve the scientific production as referential data bases like Medline, PubMed, Latin American and Caribbean Health Sciences (LILACS) and Spanish Health Sciences Bibliographic Index (IBECS) are listed. The initiative called the Virtual Health Library (VHL) developed by BIREME/PAHO/WHO, especially SciELO (Scientific Electronic Library Online) the electronic publication model, that gather collections of scientific publications- full text--, with high quality is detailed. PMID- 12884473 TI - [The effect of prebiotics on lipid metabolism]. AB - Prebiotics were defined in 1995 as non-digestible food ingredients beneficially affecting the host by stimulating the growth and/or activity of one or more bacteria in the colon, thus improving health. The proliferation of certain bacteria by fermentation of non-digestible carbohydrates has been shown to be able to inhibit the colonization of the intestine by pathogens, thus giving a protective effect vis-a-vis acute or chronic intestinal disorders. The fermentation of prebiotics may promote some specific physiological functions through the release of metabolites from the bacteria, especially short chain fatty acids (acetate, propionate, butyrate, lactate, etc.) into the lumen of the intestine. Short chain fatty acids may act directly or indirectly (by modifying the pH) on intestinal cells and may be involved in the control of various processes such as the proliferation of mucosa, inflammation, colorectal carcinogenesis, mineral absorption and the elimination of nitrogenated compounds. Curiously, numerous papers have hinted at the possibility that prebiotics may have systemic physiological effects that are related to beneficial effects on lipid metabolism and various cardiovascular risk factors. PMID- 12884474 TI - [Nutritional effect of bariatric surgery with Scopinaro operation. Analysis of 40 cases]. AB - OBJECTIVES: To analyze the influence of bariatric surgery over nutritional status in patients with morbid obesity after 1 year of following. To know the influence of specific factors as age, sex and common channel's length. DESIGN: Retrospective study from 1998-2001. SUBJECT: All patients suffering from morbid obesity that had been operated during this period of time, 40 subjects: 28 women and 12 men with a mean age of 38 +/- 12 years old. Were studied. METHODS: We analyzed the variations of weight and other anthropometric measures, biochemical parameters and the micronutrient (vitamins A, D, E, B12, folate, C, zinc, Calcium, Magnesium) status during 1 year of following after surgery. RESULTS: Percentage weight loss was 29.7 +/- 2.9% without sex differences. A significant (p < 0.005) descent of serum cholesterol, triglycerides, and uric acid level as well as liver function markers was observed. All the hyperglycemic disturbances were reverted. 97% and 48% of the patients developed hypovitaminosis A and D respectively, that was correlated to steatorrhea. Vitamin E deficiency was found in 72% of the patients, and zinc deficiency in 68% CONCLUSIONS: Our expected weight loss 1 year after bariatric surgery is 30%, irrespective of sex and age. This loss is accompanied by a significant improvement in metabolic parameters, but the high prevalence of micronutrients deficiency that was found makes an early supplementation and close follow-in of these patients very advisable. PMID- 12884475 TI - Validation of an in vitro nutrition model using an enteral formula in aged neutrophils. AB - The aim of the study is to validate a cell culture model appropriate for assessing the effects of standard nutritional formulas on neutrophil functionality in vitro. The model consists of aged cells exposed to a commercial nutritional formula containing solely LCT as lipid component. Preliminary experiments determined dosage of formula and culture interval. Neutrophils were isolated from a pool of whole blood in healthy volunteers (18-55 years old) and cultured with and without addition of a commercial enteral diet with 3.5% lipids (equivalent to 0.04, 0.08, 0.2 and 0.4 mM of intraassay LCT) for 18, 42 or 76 hours. Based on cell viability results, doses of 0.2 and 0.4 mM LCT and culture time of 18 hours were established for subsequent experiments. Neutrophil functionality was evaluated by phagocytosis (NBT test), MDA production (lipoperoxidation index) and DNA fragmentation. Optic microscopy showed higher percentages of pre-apoptotic cells and a significant increase in DNA fragmentation as compared to controls only with an LCT concentration of 0.4 mM (p < 0.05). Interestingly, cell cultures with both 0.2 and 0.4 mM of added LCT showed significant decreases in malonyldialdehyde (MDA) release as a lipoperoxidation marker. This nutrition model of neutrophils and in vitro complete enteral commercial diet is relatively simply to execute and can be applied to different pathological conditions in which the aim is to study changes in neutrophil functionality. PMID- 12884476 TI - [Caloric overload in critically ill patients treated with parenteral nutrition]. AB - OBJECTIVE: To assess the degree of hypernutrition of critically-ill patients under treatment with parenteral nutrition (PN) in a multi-purpose intensive care unit (ICU). SCOPE: Patients under treatment with parenteral nutrition in a multi purpose intensive care unit. Prospective study lasting four months. INTERVENTION: The amounts of the daily dose of glucose, lipids and nitrogen were calculated in PN, enteral nutrition (EN), dextrose solution (DS) and propofol. The daily dose of glucose and lipid administered intravenously (i.v.) was assessed with respect to the recommended value (4-5 mg/kg/min and 1.5 g/kg/day, respectively) and with respect to the dose prescribed in the PN regime The total daily calorie intake (i.v. plus EN) was assessed with respect to the recommended value (25-35 kcal/kg/day). RESULTS: The study involved 30 patients totalling 488 days with PN. The total daily dose of i.v. lipids (PN plus propofol) exceeded the recommended value on 23.2% of the days with propofol (13 of 56) and on 3.7% of the days without propofol (16 of 432). The total daily dose of i.v. dextrose did not exceed any day the maximum metabolization threshold. On 28.2% of the days with EN and 39.6% of the days without EN, the total daily dose of i.v. dextrose exceeded the PN regimen. Similarly, on 41% of the days with propofol, the total daily dose of i.v. lipids exceeded the PN regimen. The total calorie intake (i.v. plus EN) exceeded the recommended value on 46.9% of the days with EN (51 of 109) and on 5% of the days without EN (19 of 379). CONCLUSION: The glucose of dextrose solution and the propofol lipid are not routinely discounted from the PN regime. A trend towards hypernutrition of the critically-ill patient is shown, especially on days with simultaneous treatment with PN and EN. PMID- 12884477 TI - [Comparative microbiological study of fresh and heat-treated yoghurt in an in vivo animal model]. AB - In an acute assay, rats fasted for 24 h were sounded with 2 ml of fresh yogurt, sterilised yogurt or distilled water, and sacrificed at 2, 4, 8 and 24 h. They were compared with non-sounded rats. The survival of the lactic bacteria of the yogurt administered in the animals' stomach and intestines was determined, and the bacteria isolated were to the Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus profiles belonging to the strains isolated originally from the yogurt. Counts of the total number of microorganisms that were grown in lactobacilli and lactic streptococci media were also made. Analyses of the different types of resulting colonies in the lactobacilli and lactic streptococci media was performed. No L. delbrueckii subsp. bulgaricus or S. thermophilus were characterized in this acute assay. The statistical analysis of the results did not show significant differences in the log UFC. g-1, of the intestinal microbiota microorganisms of the three groups of animals in any of the times. In a chronic assay, three groups of animals, after 30 days of nourishment ad libitum with either a semisynthetic diet with 10% of fresh yogurt, or with 10% of sterilised yogurt or controls, housed in individual metabolism cells, were sacrificed. The results did not show meaningful differences in the long UFC. g-1 of the intestinal microbiota microorganisms from the three groups of animals in any of the times. In turn, in the study of the different types of colonies resulting in the lactobacilli and lactic streptococci media, no L. delbrueckii subsp. bulgaricus or S. thermophilus were characterised in this chronic assay. The results did not show meaningful differences in the log CFU. g-1 of the intestinal flora microorganisms from the three groups of animals in any of the times. In turn, no L. delbrueckii subsp. bulgaricus or S. thermophilus were characterised in this chronic assay. PMID- 12884478 TI - [Weight monitoring in parenteral nutrition]. AB - OBJECTIVE: To assess the usefulness of weight monitoring as a routine quality assurance method in the preparation of parenteral nutrient units (PUNs). MATERIAL AND METHODS: In accordance with their frequency of use, two parenteral nutrition formulations were chosen, one prepared externally (NP-I) and the other internally (NPH-2500). The relative difference between the experimental weight and the theoretical weight was used as the response variable. This difference, expressed as a percentage, defines the preparation gravimetric error (EGP in its Spanish acronym) Weight monitoring was effected on 615 PUNs, of which 337 (55%) corresponded to the NPH-2500 formulation and 278 (45%) to the NP-I formulation. The quality of the preparation of the PUNs was established by means of the calculation of the exactness (mean EGP) and precision (square root of the mean of the squares of the EGP values), and the alert and invalidation limits for these parameters were defined. The usefulness of weight monitoring was established by the time needed to detect a preparation gravimetric error of more than 5%. RESULTS: The overall prevalence of EGP in excess of 5% was less than 0.6% (95% CI: 0.0 to 0.6%). The overall exactness was 1.01% (95% CI: 0.96 to 1.05%). The difference between the exactness of NPH-2500, 0.66% (95% CI: 0.62 to 0.70%), and NP-I, 1.42% (95% CI: 1.36 to 1.49%), was statistically significant. The percentage of PUNs located outside the limits of invalidation for precision was significantly higher in the NPH-2500 formulation, 2.67% (95% CI: 2.36 to 3.00%), than in the NP-I formulation, 0.36% (95% CI: 0.25 to 0.49%). No statistically significant differences were detected in the remaining cases. CONCLUSION: Weight monitoring is a strategy that, possibly in combination with other methods, may complement the quality assurance processes defined to improve the preparation of parenteral nutrient units. PMID- 12884479 TI - Iron, folate and vitamins B12 & C dietary intake of an elderly institutionalized population in Leon, Spain. AB - Aging is associated with increased risk of developing anaemia and micronutrient deficiencies. The purpose of this study was to evaluate the daily intake of micronutrient whose deficient in diet could cause anaemia (iron, folic acid, vitamin B12) and vitamin C to establish the prevalence of anaemia in a group of institutionalized of 124 elderly subjects residing in five nursing homes in Leon (Spain). A precise weighing method was used to conduct the control of food intake covering seven days. Energy, alcohol, iron, folate, vitamin B12, and vitamin C intake were obtained. Weight, and Height also were measured. Serum iron, serum ferritin, haemoglobin and hematocrit were also measured. Average daily iron intake was higher than the 10 mg recommended by the National Academy of Science although significantly higher (p < 0.05) in males (17.0 +/- 7.4 mg) than in females (11.8 +/- 1.5 mg). Moreover, vitamin C intake in all subjects is high (118.8 +/- 43.7 mg) and higher than Spanish RDA (198%). Average intakes of folate and vitamin B12 in the present study exceeded the RDA, (103% and 144%). However, 45.83% of males and 5.97% of females showed deficiencies in vitamin B12 and 53.91% of the subjects showed deficiencies in folic acid. The average haemoglobin concentration (14.28 +/- 1.33 g/dL), hematocrit percentage (43.71 +/- 6.31), ferritin concentration (87.01 +/- 59.74 ng/mL) and serum iron (85.36 +/- 33.98 micrograms/dL) showed similar figures to the results obtained in other studies carried out on elderly populations. It would be necessary to adequately compose the menus given in nursing homes, decreasing energy contribution, and supplying micronutrient rich foods or fortified foods. PMID- 12884480 TI - [Dietetic-nutritional treatment in chronic radiation enteritis. A complex clinical case]. AB - We present a clinical case of chronic radiation enteritis, with follow-up over 34 months, the main features of which were as follows: A 60-year-old female patient with adenocarcinoma of the rectum who required a left hemicolectomy (Hartmann), receiving chemotherapy with fluorouracil and external radiation therapy (45 Gy), presenting sub-occlusive episodes five months later that required a first surgical treatment with release of adhesive bands suggesting radiation enteritis. The patient presented progressive intolerance to oral feeding and nutritional supplements, accompanied by severe malnutrition, requiring parenteral nutrition (PN), endovenous fluid therapy and requiring a second surgical intervention due to persistence of the occlusive condition, with resection of 70 cm of the terminal ileum with ileum-colon anastomosis. The pathological analysis of the segment confirmed chronic radiation enteritis, persisting with clinical manifestations of sub-occlusion and radiological alterations compatible with radiation enteritis on the remaining intestine. The dietetic and nutritional management following admission was exclusive parenteral nutrition for 3 months, receiving formula food with glutamine for four months, associated, after 79 days of intestinal rest, with oral formula diets (Basic formula with amino acids as protein source and Standard diet) and with a gradual reduction in the number of weekly infusion until nutritional autonomy was restored after 10 months with PN, with her nutritional needs being covered completely by the digestive route. Subsequently, a mixed regime was established combining diet therapy with good tolerance and normalization of the nutritional status with a return to weight prior to illness and an improvement in radiation enteritis and biochemical parameters. PMID- 12884481 TI - New JCAHO standards are here: what changes do you need to make now? AB - Now that the long-awaited revised accreditation standards from the Joint Commission on Accreditation of Healthcare Organizations have been unveiled, what changes should you make in the way you prepare for surveys? PMID- 12884482 TI - Credential volunteers during disasters. AB - After the terrorist attacks of 9/11, area hospitals all reported a deluge of volunteer clinicians. This may sound like good news when your facility is suddenly overwhelmed with patients, but it also can be dangerous. PMID- 12884483 TI - Are you complying with restraint standards? PMID- 12884484 TI - Tips from a recent survey. PMID- 12884485 TI - Don't impairments jeopardize patient safety. PMID- 12884486 TI - [Obituary to Professor/Doctor of Pharmaceutical sciences Jacek Brzezinski, 1934 2003]. PMID- 12884487 TI - [Do cells play role in the assessment of risk due to drugs and environmental factors and by what means?]. PMID- 12884488 TI - [Risk analysis rules for the establishing of maximum residue limits of veterinary medicinal products]. PMID- 12884489 TI - [Quantitative methods in the assessment of cancer risk factors]. PMID- 12884490 TI - [Evaluation of hemolytic effect of ethylene glycol ethers: risk assessment]. PMID- 12884491 TI - [Risk analysis of residues of veterinary drugs]. PMID- 12884492 TI - [Risk analysis of soil environmental pollution with heavy metals]. PMID- 12884493 TI - [Time series analysis for the evaluation of risk of death associated with environmental air pollution]. PMID- 12884494 TI - [Identification of delayed health effects of short-term changes of air pollution]. PMID- 12884495 TI - [The significance of animal studies for the assessment of prenatal risk factors]. PMID- 12884496 TI - [Health risks associated with environmental exposure to polycyclic aromatic hydrocarbons (PaHs)]. PMID- 12884497 TI - [The use of biomarkers in the assessment of health risk from environmental exposure to chemical carcinogens]. PMID- 12884498 TI - [Toxicokinetics and metabolism of 0-anisidine in rats]. PMID- 12884499 TI - [Application of cotinine determination for the assessment of active and passive smoking of delivering women]. PMID- 12884500 TI - [Application of environmental categories in estimation of active and passive smoking exposure]. PMID- 12884501 TI - [Levels of cadmium, zinc, copper and selenium in subjects with diagnosed prostate cancer]. PMID- 12884502 TI - [Liver radiosensitivity to gamma radiation in mice poisoned with N nitrosodiethylamine]. PMID- 12884503 TI - [Risk assessment for the operators of agricultural -chemical pesticides]. PMID- 12884504 TI - [Scientific background of occupational exposure limits for chemical substances in Poland]. PMID- 12884505 TI - [The content of selected contaminants in daily diets of families from Podlaskie and Silesia regions]. PMID- 12884506 TI - [Examination of students' working environment during laboratory training]. PMID- 12884507 TI - [Use of element concentration ratios in the estimate of their metabolism in the teeth of people environmentally exposed to metals]. PMID- 12884508 TI - [Comparison of auc values for caffeine and its metabolites after single and repeated administration of 2-methoxy-5-nitrobenzyl bromide]. PMID- 12884509 TI - [Hybrids of human and monkey adenoviruses (adeno-adeno hybrids) that can reproduce in monkey cells: biological and molecular genetic peculiarities]. AB - A highly oncogenic monkey adenovirus SA7(C8) facilitates the reproduction of human adenovirus type 2 (Ad2) in monkey cells. Upon mixed infection of monkey cells with both viruses, these viruses recombine producing defective adeno-adeno hybrids Ad2C8 serologically identical to Ad2 and capable of assisting Ad2 to reproduce in monkey cells. Ad2C8 and Ad2 form an intercomplementary pair inseparable in monkey cells. Unlike oncogenic SA7(C8), Ad2C8 is a nononcogenic virus for hamsters but is able to induce tumor antigens of this virus (T and TSTA). Molecular genetic analysis of 68 clones of adeno-adeno hybrids revealed that the left part of their genome consists of Ad2 DNA, and the right part contains no less than 40% of the viral SA7(C8) genome where E2A, E3, and E4 genes are located. Apparently, the products of these genes contribute to the composition of adenoviral tumor antigens, while the E4 gene is involved in complementation of monkey and human adenoviruses and makes a contribution to host range determination of these viruses. PMID- 12884510 TI - [Effect of the pho85 mutation on the catabolite repression of the CIT1 gene in yeasts Saccharomyces cerevisiae]. AB - The Krebs cycle is one of the major metabolic pathways in a cell, which includes both catabolic and anabolic reactions. The first enzyme of the Krebs cycle, citrate synthase, catalyzes one of a few irreversible reactions of the cycle, citrate formation from acetyl-CoA and oxaloacetate. Expression of the CIT1 gene encoding the mitochondrial form of this enzyme in Saccharomyces cerevisiae is repressed on glucose- and glutamate-containing medium and activated on the raffinose-containing medium. In this work, the dependence of glucose repression of the CIT1 gene on the content of phosphate in the medium was studied. On the phosphate-deficient medium, the level of the CIT1 gene expression was increased twice. A low-molecular-weight (about 34 kDa) protein was identified and shown to interact with a region of the CIT1 gene promoter (from -367 to -346 bp), which controls the glucose repression. The results obtained suggest that the Pho4 protein is involved in regulation of the CIT1 gene expression on the glucose containing and phosphate-deficient medium. Disruption of the PHO85 gene encoding phosphoprotein kinase (Pho4p is the substrate of this enzyme) leads to alleviation of glucose repression of the CIT1 gene. Thus, in yeast cells grown in the presence of glucose, the PHO85 gene mediates downregulation of the CIT1 expression. PMID- 12884511 TI - [The genetic control of cell growth and development in yeast Saccharomyces cerevisiae. Disturbed sporulation in diploids with decreased activity of the Ras/cAMP signal transduction pathway]. AB - Seven haploid strains (four with the MAT alpha mating type and three with the MATa mating type) were selected from the Peterhof genetic collection of yeast. Previous phenotypic analysis assigned six of these strains to a physiological group of strains with a lower activity of the Ras/cAMP signal transduction pathway. The haploids were crossed, and the resulting 12 diploids showed higher glycogen accumulation, tolerance to heat shock and nitrogen starvation, and sporulation in complete media. Ten of the diploids expressed the hypersporulation phenotype (higher sporulation efficiency). The phenotypic characters of these ten diploids suggested a reduced activity of the Ras/cAMP pathway. All 12 diploids were tested for sporulation and production of two groups of asci (those with one or two spores and those with three or four spores) as dependent on culture conditions (21, 30, or 34 degrees C; standard sporulation medium or a complete medium containing potassium acetate or glycerol in place of glucose). Sporulation proved to depend on temperature and medium composition. The results are collated with the data on yeast phenotypes associated with a lower activity of the Ras/cAMP signal transduction pathway. PMID- 12884512 TI - [Effect of insertional mutation in the cspA gene encoding the major cold-shock protein on radiation resistance of Escherichia coli]. AB - Plasmid pCspA::Km carrying a cloned mutant allele of the cspA gene for the major Escherichia coli cold-shock protein CspA with an insertion of the kanamycin resistance gene cassette from transposon Tn903 into the core region of the coding sequence causes a 2.3-fold increase in radioresistance of wild-type E. coli cells (cspA+). The radioprotective effect of this plasmid is abolished or drastically reduced in mutants recA13 and rpoH15 defective in RecA protein and in induction of the heat-shock protein regulon, respectively. Plasmid pCspA::Km causes a 1.3 fold elevation in the resistance to gamma-irradiation of E. coli mutants with an intermediate level of radioresistance (Gamr445 and KS0160) but slightly diminishes resistance of a highly radiation-resistant Gamr445 mutant. In the chromosome of E. coli with normal DNA repair systems, the cspA::Km mutation in the homozygous state enhances resistance to the lethal effect of gamma-rays and UV light 2.9 and 1.4 times, respectively. These data suggest that the system of cold-shock proteins can modulate resistance of E. coli cells to the lethal effect of gamma-rays and UV light. PMID- 12884513 TI - [Loach spermatozoa transfer foreign DNA, which expression is discovered in the early development stages]. AB - The transfer of plasmid pcDNA3-lacZ by electrotrasfected sperm cells into loach (Misgurnus fossilis L.) ova has been studied. The lacZ gene has been found to express in 3- to 5-day-old prelarvae. PMID- 12884514 TI - [Mitochondrial DNA polymorphism in the natural populations of the Drosophila virilis species group]. AB - Restriction fragment length polymorphism (RFLP) analysis has been used to evaluate mitochondrial DNA (mtDNA) variation in 12 sibling species forming the Drosophila virilis species group. The variation thresholds corresponding to the interspecific and interstrain levels have been determined. The results indicate that interspecific hybridization has significantly contributed to the evolutionary history of the virilis species group. PMID- 12884515 TI - [The frequency of reversion of the opaque2 locus mutable alleles controlled by the Bg-rbg system of mobile elements in maize]. AB - Maize lines differing in the frequency of reversion of the opaque2 (o2) mutable alleles controlled by the system of Bg-rbg transportable elements were studied. In the presence of the Bg regulatory element, these alleles can revert to normal. When reversion occurs prior to the first division of the primary endosperm nucleus, either phenotypically normal kernels or whole endosperm revertants (WER) develop. Low frequency of whole endosperm revertant formation may be produced by different genetic mechanisms. The frequency of WER formation was shown to nonlinearly depend on the dose of the Bg-hf regulatory element. A dose increase from two to three failed to cause an essential increase in the number of revertants. The regulatory elements Bg-lf and Bg-hf differed in ability to induce excision of the receptor element at the same dose. The frequency of reversion of the receptive alleles was shown to be regulated by epigenetic mechanisms so that high frequency of reversion of receptive alleles requires preliminary premeiotic association between the regulatory and receptor elements. The inheritance of the maize alleles o2-hf and o2-lf proved to be similar to that an3 mutable alleles in petunia. PMID- 12884516 TI - [Genetic analysis of inheritance of mei8, sy1 and sy10 mutations, disrupting the correct meiosis in the rye Secale cereale L]. AB - It is shown that mutations mei8 (irregular condensation and fragmentation of meiotic chromosomes), sy1 (asynapsis), and sy10 (heterologous synapsis) of rye Secale cereal are nonallelic. In double mutants mei8 sy1 and mei8 sy10 both mutations are expressed simultaneously and independently of each other. A study of joint inheritance of mutations sy1 and sy10 revealed their interaction by means of recessive epistasis: the double mutants has the sy10 phenotype. This means that the sy10 gene controls an earlier stage of synapsis in meiotic prophase than the sy1 gene. Mutation mei8 is inherited independently of sy1 but it is linked to sy10 (recombination frequency 26.8 +/- 3.58%). PMID- 12884517 TI - [The study of joint inheritance of mutations impairing the structure of meiotic chromosomes in rye Secale cereale L]. AB - Genetic analysis has demonstrated that meiotic mutations mei8 (irregular condensation and fragmentation of meiotic chromosomes) and mei10 (chromosome overcompaction) are nonallelic. Mutation mei10 exhibits digenic inheritance (with a segregation ratio of 13:3) in the combinations of crosses studied. It is assumed that the phenotypic expression of mutation mei10 is suppressed by the effect of recessive gene lch1 or lch2 (long chromosomes), both of which have been revealed in one of the parental lines (Mc10). These genes determine weak condensation of meiotic chromosomes. In double mutants mei8 mei10, the mutations are expressed independently of each other. Gene mei10 is linked with gene mei8 (r = 36.8 +/- 5.38%); genes lch1 and lch2 are not linked either with them or with each other. Taking into account the data on the linkage between genes mei10 and sy10 and between mei8 and sy10, the order of genes in the linkage group is shown to the following: mei8-sy10-mei10. PMID- 12884518 TI - [RAPD-based analysis of introgression of barley genetic material into the genome of alloplasmic wheat lines (Hordeum geniculatum All./ Triticum aestivum L.)]. AB - Genomes of three alloplasmic wheat lines obtained on the basis of barley--wheat hybrid Horderum geniculatum All. (2n = 28) x Triticum aestivum L. (2n = 42)(Pyrotrix 28) were examined using random amplified polymorphic DNA (RAPD) analysis. Line L-29 was obtained after first backcross of the initial hybrid with the wheat variety Pyrotrix 28 and ten subsequent self-pollinating generations. This line was represented by euploid plants with typical to the common wheat chromosome number (2n = 42), as well as by aneuploids, which contained an additional telocentric chromosome in the main karyotype (2n = 42 + t). Lines L-26 and L-27 were obtained by two backcrosses of one BC1 plant with the wheat variety Novosibirskaya 67 and one subsequent self-polination of one BC3 plant. Chromosome number in all these plants corresponded to 2n = 40 + 4t. RAPD analysis was carried out using seven primers, which were previously proved to be effective for identification of the barley genome fragments within hybrid genomes of alloplasmic lines. The presence of barley genome fragments in line L-29 was revealed by use of five primers, while in lines L-26 and L-27 these fragments were detected by use of one primer. The significant difference in the number of barley RAPD fragments in the genomes of alloplasmic lines obtained at different backcrossing stages suggests more intense displacement of barley genome during backcrossing compared to self-pollination in BC1 plants. PMID- 12884519 TI - [Variation of the cytoplasm type in sugar beet (Beta vulgaris L.) upon inbreeding. The influence of hybridization]. AB - Hybrid combinations of inbred sugar beet lines that undergo conversion of N cytoplasm into S-state were screened for the marker mitochondrial genes atpA and atp6. The involvement of nuclear factors into cytoplasm conversion and possible identity of these factors in different lines have been studied. The cytoplasm conversion factor was localized to nucleus. In different lines with cytoplasm conversion, the nuclear conversion factors are not identical. The state of the mitochondrial genome is normalized after outcrosses with plants having the stable cytoplasm. PMID- 12884520 TI - [Oligodontia and its inheritance in Kerry blue terrier]. AB - Defects of the premolar tooth formula (oligodontia, tooth number reduction) were studied in dogs of the Kerry blue terrier breed. For this purpose, a database including 480 individuals of 96 litters was constructed. The occurrence of oligodontia was investigated in pedigree groups with inbred and outbred crosses. No selective mating choice for the anomaly under study was found in the sample. The results indicate that oligodontia is inherited, which requires comprehensive study of its genetic control and search for corresponding genes. PMID- 12884521 TI - [A search for genetic loci responsible for emotional stress-induced arterial hypertension in the ISIAH rats]. AB - Hypertension is a widespread human disease caused by a complex interaction of a series of the genetic factors with both each other and the environmental conditions. In this study we aimed at determining the candidate genetic loci responsible for hypertension in the ISIAH rats and studying the dynamics of the relevant genetic and physiological mechanisms in rat ontogeny. The candidate genetic loci were identified from association of the microsatellite markers linked to these loci with arterial hypertension in rat F2 hybrids exposed to stress. Two populations of F2 hybrids of different age (3-4 and 6 months) were obtained by crossing hypertensive ISIAH and normotensive WAG rats. We present the results of cosegregation analysis for the following loci: the gene for the Na+, K(+)-ATPase alpha 1 subunit isoform (Atp1a1), the endothelin-2 gene (Edn2), the low affinity nerve growth factor receptor gene (Lngfr), and a region of chromosome 10 marked with the D10Rat58 microsatellile located 3 cM away of the aldolase C gene (AldC). The results obtained allowed us to localize the genes responsible for the stress-induced arterial hypertension in the ISIAH rats to the Atp1a1 locus (P < 0.05), chromosome 2 and to the Lngfr gene locus (P < 0.05), chromosome 10. The association of hypertensive status with the Lngfr gene was found only in young ISIAH rats whereas in adult rats of this line, hypertension was associated with the Atp1a1 locus. PMID- 12884522 TI - [Comparative compositional mapping of chicken and quail chromosomes]. AB - The distribution of various isochore families on mitotic chromosomes of domestic chicken and Japanese quail was studied by the method of fluorescence in situ DNA- DNA hybridization (FISH). DNA of various isochore families was shown to be distributed irregularly and similarly on chromosomes of domestic chicken and Japanese quail. The GC-rich isochore families (H2, H3, and H4) hybridized mainly to microchromosomes and a majority of macrochromosome telomeric regions. In chicken, an intense fluorescence was also in a structural heterochromatin region of the Z chromosome long arm. In some regions of the quail macrochromosome arms, hybridization was also with isochore families H3 and H4. On macrochromosomes of both species, the pattern of hybridization with isochores of the H2 and H3 families resembled R-banding. The light isochores (L1 and L2 families) are mostly detected within macrochromosome internal regions corresponding to G bands, whereas microchromosomes lack light isochores. Although mammalian and avian karyotypes differ significantly in organization, the isochore distribution in genomes of these two lineages of the warm-blooded animals is similar in principle. On macrochromosomes of the two avian species studied, a pattern of isochore distribution resembled that of mammalian chromosomes. The main specific feature of the avian genome, a great number of microchromosomes (about 30% of the genome), determines a compositional specialization of the latter. This suggests the existence of not only structural but also functional compartmentalization of the avian genome. PMID- 12884523 TI - [The correlation between brain weight and behavior changes in response to ethanol administration in laboratory mice]. AB - The effects of ethanol injections on the F2 offspring of the cross between large brain (LB) and small-brain (SB) mouse strains selected for high and low relative brain weights, respectively, have been studied. The parental strains have significantly differed in brain weight for many generations. The effects of ethanol (2.4 g/kg) have been compared in four subpopulations of mice that differ pairwise in brain weight. One pair of subpopulations has been isolated from the hybrid group and the other, from generation 22 of selection of the parental strains. The results of ANOVA have demonstrated that brain weight is related to the response to ethanol injections. The parameters of stereotyped behavior, which increased in after ethanol injections and reflected the decrease in exploratory activity) were different in mice with high and low relative brain weights. The pattern of behavioral changes after ethanol injections is the second (after increased learning ability) behavioral trait found to be correlated with brain weight. PMID- 12884524 TI - Determination of the CCR2-64I/CCR5-59653T haplotype linkage disequilibrium in a sample of Polish population. AB - It has been reported that different polymorphisms in the regulatory regions of CCR5 and in the CCR2 gene of the chemokine receptors are associated with AIDS. We decided to determine the congruence between CCR5-59653T and CCR2-64I alleles in a group of 281 persons. Frequencies of combined CCR5-59653T and CCR2-64I haplotypes were examined in a group of 281 persons. Among 281 individuals 26 (9.3%) and 24 (8.5%) respectively were carriers of the CCR5-59653T and the CCR2-64I alleles. We also found that 24 persons (8.5%) were carriers of combined CCR2-64I/CCR5-59653T allele. The calculation of the congruence revealed that 92.0% of individuals exhibited the same genotype for both CCR2-64I and CCR5-59653T polymorphisms. Our results confirm that linkage between CCR5-59653T and CCR2-64I alleles is not absolute. PMID- 12884525 TI - Correlation of age and birth order of parents with chromosomal anomalies in children. AB - One hundred children with suspected congenital and/or malformation and their parents who reported to SAT hospital, Medical College, Trivandrum, India formed the test group. Fifty children with no obvious anomalies or abnormalities and their parents formed the control group. The criteria for selection of the control was 1) the maternal age at delivery was below 30 years and 2) the parents belong to 1st or 2nd birth order. The chromosomal analysis was carried out in all the subjects using peripheral blood lymphocyte microculture to investigate for any constitutional chromosomal markers and quantitate the mutagen (bleomycin) sensitivity of the chromosomes. All the subjects were evaluated clinically and a complete family history was recorded. Chromosome anomalies were noted in 41 out of the one hundred children and in 4 out of the 200 parents of the test group. No constitutional aberrations were seen either in the parents or in the children of the control group. Bleomycin sensitivity study revealed a high b/c value in 35 children (24 hypersensitive and 11 sensitive) of the test group whereas in the control group the b/c values were low denoting hyposensitivity and very good DNA repair mechanism. This study reveals that the incidence of chromosome aberrations is higher when the age and birth order of parents are higher. A direct correlation was noted with parental order and b/c value. This was also true with the parental age and birth defects. PMID- 12884526 TI - [Correlation between polymorphic variants of the genes for transferrin and angiotensin-converting enzyme and antioxidant activity of blood plasma]. AB - In 75 male and 46 female subjects of an urban population (93% Russians) and in 38 males and 40 females of a rural population (87% Russians), the antioxidant activity (AOA) of blood plasma was determined from the plasma ability to reduce the yield of products interacting with thiobarbituric acid in the model lecithin Fe2+ ion system. In the urban population, the loci TF (AvaI in exon5) and ACE (I/D polymorphism of the Alu repeat in intron16) were studied in 130 and 141 subjects, respectively. Of them, 102 and 111 subjects, respectively, were examined for AOA. In the rural population, the corresponding sample sizes were 75 and 76 (73 and 74 subjects were examined for AOA). The polymorphic loci of the urban and rural populations did not differ in the allele frequencies. In both populations Hardy--Weinberg and gametic equilibria were observed. The contributions of the TF and ACE genes to AOA variation in the combined sample from the urban and rural populations were 0.6 and 0.5%, respectively. PMID- 12884527 TI - [Analysis of mutations in the RET proto-oncogene in patients with medullary thyroid tumor]. AB - The spectrum of mutations of the RET protooncogene was analyzed in Russian patients with inherited or sporadic medullary thyroid carcinoma (MTC). Four RET exons (11, 13, 15, and 16) were subjected to molecular analysis, and mutations were revealed and identified in 47.4% (9/19) patients with sporadic MTC. In total, six mutations (including three new ones) were observed. The most common mutation affected codon 918 to cause substitution of methionine with threonine and accounted for 31.6% alleles. Analysis of exons 11 and 16 revealed four mutations in patients with inherited multiple endocrine neoplasia type 2 (MEN 2). Mutations were found in each patient. Thyroidectomy was performed in four asymptomatic carriers of RET mutations from three MET 2A families (in two families, affected relatives had bilateral pheochromocytoma). In two patients, analysis of the surgery material revealed MTC microfoci in both lobes of the thyroid gland. The results provide the ground for constructing a bank of genetic information on Russian MTC patients with the clinically verified diagnosis. PMID- 12884528 TI - [Analysis of the N-acetyltransferase 2 gene polymorphism in the patients with chronic obstructive pulmonary disease and in populations of the Volga-Ural region]. AB - Restriction fragment-length polymorphism of the gene coding for N acetyltransferase 2 (NAT2) was typed in populations of the Volga-Ural region (Bashkirs, Tatars, Chuvashes, Udmurts, and Russians) as well as in patients with chronic obstructive pulmonary disease (COPD) and in healthy individuals. Rapid and slow acetylator phenotypes were determined based on the presence or absence of the KpnI, TaqI, and BamHI restriction endonuclease recognition sites. The proportion of slow acetylators in the populations examined varied from 40.00% in Bashkirs to 64.15% in Chuvashes with statistically significant difference between these two ethnic groups (chi 2 = 5.7; p = 0.02). Overall, in the Volga-Ural populations slow acetylators represented 56.25% of the subjects examined. This value was similar to those presented in other studies of Caucasoid populations. In the COPD patients a statistically significant decrease of the slow acetylator frequency to 48.28% compared to healthy individuals (62.18%) was observed (chi 2 = 4.60; p = 0.036). The data obtained suggest a possible association between the drug resistance in the COPD patients with the rapid acetylator phenotype, which can lead to the development of the chronic form of the disease. PMID- 12884530 TI - Hot words. A claim of nonhuman-induced global warming sparks debate. PMID- 12884529 TI - [A search for mutations in the DIA1 gene in case of hereditary methemoglobinemia type I in the iakut population]. AB - In the patients with enzymopenic hereditary methemoglobinemia type I, a disease widely distributed on the territory of Yakutia, a search for the mutations in exons 3 and 4 of the DIA1 gene encoding NADH-cytochrome b5 reductase was carried out. It was shown that Yakut patients have none of three missence mutations, Arg57Gln, Leu72Pro, and Val105Met, described in case of this disease in the neighboring populations, Chinese and Japanese, inhabiting the territories south of Yakutia. PMID- 12884531 TI - Secret ingredients. PMID- 12884532 TI - Sourcing sapiens. New fossils and DNA tests get to the roots of our species. PMID- 12884534 TI - Energy crunch. PMID- 12884533 TI - View from VIRGO. PMID- 12884535 TI - Hands of light. Moving particles with photons leads to a new form of nanomanufacturing. PMID- 12884537 TI - Demystifying the digital divide. PMID- 12884536 TI - Censors of the genome. PMID- 12884538 TI - Rethinking the "lesser brain". PMID- 12884539 TI - Information in the holographic universe. PMID- 12884540 TI - Questioning the Delphic oracle. PMID- 12884541 TI - Planet of the apes. PMID- 12884542 TI - Night vision. Seeing green. PMID- 12884543 TI - Virologic and serologic investigations of West Nile virus circulation in Belarus. AB - In 1985-1994 virologic and serologic investigations were performed for the purposes of West Nile (WN) virus circulation establishment on the territory of Belarus. Blood-sucking mosquitoes, midges, wild small mammals, birds as well as blood and cerebrospinal samples from patients with nondifferentiated fevers and from healthy individuals were under studies. Four virus strains were isolated in Belarus for the first time, namely: 1--from birds (48-WN Tremlya); 2--from Aedes mosquitoes (319 and 2438); 1--from a febrile patient (Win). Their antigenic and biological properties were examined in cell cultures and laboratory animals. The isolates turned to be identical with each other and closely related to reference Egypt strain Eg 101, that is a topotype for the African virus group. One more WN virus strain (8891) was isolated from Anopheles mosquitoes in 1999. Specific antibodies to the virus in human blood sera were identified by immunological and serologic assays in 1.7% of Belarusian population. In Gomel and Brest Regions the percentage of seropositive individuals reached 5.8 and 15.4, respectively. WN virus antibodies prevailed in 0.6-5.8% of cattle, in 2.9-6.8% of wild small mammals and in 6.5-16.7% of birds. Thus, the conclusion was made on the existence of favourable conditions for the virus spread throughout the whole country and in the south in particular. Blood-sucking mosquitoes and birds are principle vectors in WN virus circulation in Belarus. 16 serologically confirmed cases of WN encephalitis were revealed in patients with fever of obscure etiology. In the view of the given data, reports on the reemergence of the pathogen in different countries and the tendency in global warming WN virus monitoring should become a subject of concern for Belarusian public medical care services. PMID- 12884544 TI - WHO publishes guidance to minimize terrorist threats to food. PMID- 12884545 TI - Ascorbic acid and infertility treatment. AB - AIM OF THE STUDY: To assess the ascorbic acid (AA) in the follicular fluid in women treated by in vitro fertilization and embryonic transfer levels (IVF/ET) and to analyse the influence of vitamin C supplementation on the results of infertility treatment. TYPE OF THE STUDY: Prospective study in women treated by IVF/ET. METHODS: The influence of vitamin C supplementation on the outcome of infertility treatment in the assisted reproduction programme in 76 women (38 of them smokers, 38 non-smokers) was studied. Half the women (19 smokers and 19 non smokers) were administered vitamin C in daily doses of 500 mg in so-called pellets allowing for gradual release over 8 to 12 hours. The control group consisted of the same number of smokers and non-smokers. In all the women, ascorbic acid levels were determined in two urine samples (prior to supplementation and at follicle retrieval) and in follicular fluid by means of a colorimetric method. Ovarian response to hormonal stimulation with gonadotropins (hMG, FSH) at a dosage of 150-225 IU per day combined with GnRH analogues in the short (buserelin) or long (triptorelin) protocols, and 5,000-10,000 IU of human chorionic gonadotropin was evaluated based on the number of follicles created and number of retrieved oocytes. Fertilisation was assessed, based on the number of successfully fertilised oocytes (fertilisation rate) and based on the number of cultivated embryos. The success of the infertility treatment was evaluated based on the number of pregnancies. RESULTS: Ascorbic acid levels in follicles were significantly higher (p < 0.001) in women with vitamin C supplementation than in the control group (8.98 +/- 5.09 vs. 5.04 +/- 2.85 mg/l). The administration of vitamin C during the period of hormonal stimulation showed a statistically insignificant impact in terms of the higher number of pregnancies (34.2% vs. 23.7%). Vitamin supplementation had a greater impact on the number of pregnancies in the non-smokers' group (57.9% vs. 31.6%). The pregnancy rate was significantly higher (p < 0.01) in non-smoking women than in smokers--44.7% vs. 13.2%, which appears to be a reason for asking women to cease smoking prior to infertility treatment. PMID- 12884546 TI - WHO calls for strong focus on health promotion for women. PMID- 12884547 TI - Effectiveness of chlorine, organic acids and UV treatments in reducing Escherichia coli O157:H7 and Yersinia enterocolitica on apples. AB - This study assessed the effectiveness of 200 and 500 ppm of chlorine and organic acids (0.5% lactic acid and 0.5% citric acid) in wash solutions, and UV radiation for reducing Escherichia coli O157:H7 and Yersinia enterocolitica on apples contaminated by two different methods. Residual levels of these pathogens after different treatments were compared. On dip inoculated apples, Y. enterocolitica reductions of 2.66 and 2.77 logs were obtained with 200 and 500 ppm chlorine combined with 0.5% lactic acid, respectively. The E. coli O157:H7 population decreased 3.35 log with 0.5% lactic acid wash solution, and 2.72 and 2.62 logs after 500 ppm chlorine and 500 ppm chlorine plus 0.5% lactic acid treatments, respectively. Similar reductions were obtained with UV radiation. On spot inoculated apples, significant (p < 0.05) decreases of 4.67 and 4.58 logs were observed in E. coli O157:H7 and Y. enterocolitica levels, respectively, after 500 ppm chlorine plus 0.5% lactic acid treatment as compared with the control. In sectioned apples, microorganisms infiltrated in inner core region and pulp were not significantly (p < 0.05) affected by disinfection treatments. No pathogens were detected in the natural microflora on apples. Reductions such as those obtained with 500 ppm chlorine plus 0.5% lactic acid solution were very proximal to the 5-log score required by FDA for apple disinfection. PMID- 12884548 TI - WHO health briefing for Iraq. PMID- 12884549 TI - Lifestyle and socio-economic status--verification of data reliability from questionnaire surveys. AB - The study of reliability was part of a project funded by the Grant Agency of the Czech Ministry of Health. The repeatability of answers, 180 questionnaires being sent again after a six week interval, was assessed by two methods: by the percentages of agreement and using the Kappa index. We evaluated 61 questions, which were divided into five sections (general questions, questions on employment, lifestyle, health, and personality). The agreement rate varied from 46% to 100% and the value of the Kappa index from -0.01 to 1. The agreement performed by the Kappa index was divided into 4 groups (< or = 0.4--poor; 0.41 0.6--average; 0.61-0.80--good; 0.81-1 almost perfect). The agreement across our questionnaire was poor in 6.6% of the questions, average--in 31.1%, good--in 45.9% and almost perfect--in 16.4% of the questions. Next we divided the questions by their content into two groups--the factual questions (group 1) and questions where answers contain an evaluative or motivational element (group 2). Significant differences (p < 0.001) were found between groups 1 and 2 in the agreement (86.8%; 72.1%) and the Kappa index (0.73; 0.48). In questions where agreement of answers is evaluated as average or even weak we used three types of solution: a) substitution by the analogous questions with a higher agreement, b) aggregation of the answers if possible, or c) the questions were not used for the analyses. These important results will be considered in further analyses of the data and in the interpretation of the study results. The detailed information about the whole project and the results has already been published or presented are available on the following web site: www.zuova.cz/projekty/ses.php. PMID- 12884550 TI - Evaluation of the response of a questionnaire study by using the GIS and standard statistical methods. AB - This study is a part of a larger project Nr. NJ 6139-3 funded by the Grant Agency of the Czech Ministry of Health. The aim of the paper was to analyse the response rate using standard statistical methods and the Geographical Information System (GIS); to indicate differences in the response by sex, age, education, and employment; to determine the breakpoint for the collection of questionnaires according to which we can estimate the total response rate; to indicate whether the study sample was representative enough to generalize the project results. The additional aim of the paper was to collect those background literary sources dealing with the response rate as a methodological paradigm. The statistical and GIS analysis were based on comparison of the total population data (Census 2001), the study sample and the sample of the completed questionnaires data in the 23 districts of the city of Ostrava. The information from the data collection was derived from the date of receipt for each questionnaire. The literature sources were obtained from the Internet--in total 228 papers from the period since 1986 to the present have been checked. The main results of this study are: the GIS analysis was confirmed in all stages by standard statistical methods--it can therefore be used as a valid tool for quick orientation in data and for the comparison of a study sample with the general population; we did not find significant differences in the course of the collection of the questionnaires between sex, age, education, and the employment of respondents; it can be seen that the breakpoint according to which we can estimate the total response rate, is the 10th day after the questionnaires are distributed by post (75% of the questionnaires collected); our sample is representative enough from the geographical point of view. More detailed information about the whole project and results already published or presented are available on the following web site: www.zuova.cz/projekty/ses/php. PMID- 12884551 TI - World Health Organization inspects Russian smallpox laboratory. PMID- 12884552 TI - Dynamics of transitory morbidity in children aged 0 to 6 years from the town of Varna. AB - Transitory chronic morbidity of children aged 0-6 years from the town of Varna was retrospectively studied for a five-year period (1997-2001) based on the results obtained during the annual prophylactic examinations. A total of 54,068 children (24,204 being 0-3 year-old and the rest of 29,864 aged between 4-6 years) is involved in the study. Based on data from the documentation records the frequency of cases per 1000 examined is given. The structure and the dynamics of transitory morbidity is determined. The total chronic transitory morbidity for the whole period is 36 per 1000 examined children. The prevalence of chronic diseases in children aged 0-3 years is 23%@1000, while in children aged 4-6 years it is twice as high (47%@1000) (p < 0.01). In the nosologic structure of chronic morbidity in children aged 0-3 years diseases of the blood and haemopoiesis dominate (52.2%) followed by congenital anomalies (7.5%), and bronchial asthma (4.1%). For the age group of 4-6 years, diseases of the blood and haemopoietic organs occupy the first place (14.3%). Second come disorders in refraction, accommodation and vision (12.8%) followed by bronchial asthma (11.8%) and scoliosis (11.2%). The results obtained in this study define the priority guidelines of the work of general practitioners and medical specialists for disease prevention and health promotion in children aged up to 6 years. They are offered to the municipal authorities of public health. PMID- 12884553 TI - 10 million Euros needed now to prepare for deadly new strain of meningitis in Africa. IFRC, MSF, UNICEF and WHO launch emergency appeal. PMID- 12884554 TI - Cancer occurrence among radiation workers at Jaslovske Bohunice nuclear power plant. AB - The Public Health Institute in Trnava, Slovak Republic under coordination of the International Agency of Cancer Research (IARC) at Lyon, France, had completed a retrospective cohort study of cancer occurrence among nuclear workers at Jaslovske Bohunice, Slovak Republic, as part of a multicentric cohort study of 14 countries. The objective of the study was to assess an impact of a long-term low level ionising radiation on cancer occurrence. In summary, 2776 employees had been selected for the follow-up period, which lasted since January 1, 1973 till December 31, 1993, it means for 21 years. Mean age at beginning of follow-up was 28.9 years and at the end of follow-up 39.6 years. The total number of person years was 27,742.1 of males and 2,442.2 of females. Exposure was expressed as annual cumulative dose of each cohort member. The total cumulative external gamma dose over the follow-up time was 58,187.9 mSv. The average external gamma dose per person years was 2.06 mSv of males and 0.37 of females. Socio-economic status, described by the last job description and education were used as possible confounding variables. All cause mortality, cancer caused mortality and cancer incidence were assessed comparing with general Slovak population using indirect standardised mortality ratio or incidence ratio calculations. There were 47 deaths reported, 44 males and 3 females, over 21 years of follow-up. The most frequent causes of death were cancer (about 30%), accidents, suicides and other external causes (about 26%), cardiovascular diseases (23%). Standardised mortality ratios of 0.39 (0.386-0.392) and 0.46 (0.27-0.59) for males and females, respectively were found in case of all cause mortality. In case of cancer mortality the standardised mortality ratios (SMR) were 0.44 (0.42-0.47) and 1.35 (0.84-1.86) for males and females, respectively. There were 32 incident cases of cancer observed during follow-up. Standardised incidence ratios as 0.508 (0.49-0.52) and 0.905 (0.74-1.06) for males and females, respectively were found. Dose response relation was measured using Poisson regression and Cox proportional hazard model. The estimated excess risks in both approaches and both for mortality and morbidity study were rather anecdotal due to lack of statistical significance caused by small number of cases. PMID- 12884555 TI - Thyroid cancer morbidity in Opole province, Poland, after Czernobyl disaster. AB - The Czernobyl disaster caused an extensive pollution in Europe due to the release into the environment thousands of tons of radioactive isotopes affecting the elevated cancer morbidity in the continent. The authors used the data set on radiation in Opole province, Poland, to model thyroid cancer incidence in the 1994-1998 quinquennium. An increase in thyroid cancer morbidity in females was observed within the 137 + 134Cs elevated concentration areas. The changes of the cancer incidence in males were found not significant for the distinguished radiation levels in the region. The statistical modeling was performed via the BUGS software. PMID- 12884556 TI - WHO releases first global reference guide on safe and effective use of essential medicines. Guide promotes consumer rights and patient safety. PMID- 12884557 TI - Serological markers of Chlamydia pneumoniae, cytomegalovirus and Helicobacter pylori infection in diabetic and non-diabetic patients with unstable angina pectoris. AB - The possible role of inflammation in coronary artery disease (CAD) is being recognised, while markers of inflammation (e.g., CRP) and infection with Chlamydia pneumoniae (C. pneumoniae), cytomegalovirus (CMV) and Helicobacter pylori (H. pylori) have been proposed as risk factors for CAD. However, these associations require further evaluation. It is a known fact that diabetic patients suffer from impaired immune response to some pathogens and a high incidence of atherosclerosis. In this case-control study we investigated serological markers of infection with C. pneumoniae, CMV, and H. pylori in a group of 140 patients with unstable angina pectoris (UA), 52 of them having type 2 diabetes mellitus, and in a matched control group. Anamnestic (IgG) and acute infection (IgA) antibodies against the above agents were tested using ELISA or indirect immunofluorescence tests. In patients with UA we found a significantly higher seroprevalence and titres of IgG antibodies against C. pneumoniae (p = 0.04) and increased titres of IgG antibodies against CMV (p = 0.007). No differences were found in IgA antibody response to these pathogens. Antibody response to H. pylori was similar in both groups tested. In diabetic patients with UA, the frequency of group-common IgG antibodies against C. pneumoniae was higher than in the non-diabetic UA patients. The other serological markers studied were comparable in the patients with or without diabetes mellitus. Our findings confirmed association of C. pneumoniae and CMV with cardiovascular heart disease. Moreover, diabetes mellitus may predispose the patients to C. pneumoniae infection. However, serological markers observed do not indicate that destabilisation of angina pectoris is associated with acute C. pneumoniae or CMV infection. No relationship was found between UA and H. pylori infection. PMID- 12884558 TI - WHO Atlas maps global tobacco epidemic. PMID- 12884559 TI - Effect of iodine and selenium upon thyroid function. AB - Iodine is an essential element with unique role in organism: it is indispensable component of thyroid hormones. After binding with specific nuclear receptor, T3/T4 induce transcription of genetic code via mRNA and regulate proteosynthesis in most tissues. Thyroid hormones regulate rate of metabolic processes and consequently development of organism. Czech Republic was in past typical region with moderate to severe iodine deficiency. Therefore epidemiological survey was started in randomly selected samples of peoples. Thereafter a complex program of improving iodine supply was realized. Prompt effects of this changes were recorded, namely increase of ioduria and decrease of thyroid volume. Essential trace element selenium has a fundamental importance to the cell and body metabolism regulation by thyroid hormones. Activities of selenoenzymes deiodinases lead to the activation of prohormone T4 to active hormone T3 and the inactivation of T3 and of T4, as well. There is unfortunately moderate to mild Se deficit in the CR. Average serum Se concentrations for the populations from 6 to 65 years are in the regions of the CR between 42 and 62 micrograms/l and urine Se values are between 8 and 15 micrograms/l. We have found statistically significant correlations among indexes of selenium status and indexes of thyroid hormone metabolism and function. Especially dangerous are concomitant deficiencies of both key elements for thyroid hormone metabolism--I and Se--from the point of thyroid hormone regulative functions. PMID- 12884560 TI - International Conference on Indoor Environment Quality in Hospitals. Prague 10-11 October 2002. PMID- 12884561 TI - [The influence oc glycocorticoid therapy on sputum ECP concentration in symptomatic patients with bronchial asthma]. AB - ECP released from the granules of activated eosinophils is regarded to be a marker of airway inflammation in asthma. The study was performed to compare the usefulness of measuring serum and sputum ECP for monitoring the asthma treatment. 29 subjects with mild to moderate asthma (mean age 41 +/- 17) were admitted in exacerbation (FEV1 55.54 +/- 87.49% N). 10 subjects with grass pollen asymptomatic asthma and 10 healthy subjects were also enrolled in the study. Patients with symptomatic asthma were ordered 30 mg prednisone for 2 weeks and they continued during next 2 weeks inhaled budesonide therapy. The concentrations of ECP (mcg/L) were determined by CAP-system (Pharmacia). The total eosinophil count and serum ECP in all subjects treated orally and next by inhaled GKS didn't differ statistically. The highest sputum ECP concentration was determined in exacerbation of asthma 84.5 +/- 78 mcg/L and statistically were reduced after 2 weeks of prednisone treatment 24.4 +/- 12.1 mcg/L (p = 0.05). In following 2 weeks of budesonide treatment sputum ECP concentration was statistically negligible in relation to previous treatment in spite of increasing tendency (50 +/- 61.3 mcg/L (p = 0.2394). In asymptomatic grass pollen asthma sputum ECP concentration was 19.7 +/- 9.4 mcg/L, higher than in controls 12 +/- 5.8 mcg/L (p = 0.04). There were a significant correlations between total eosinophil count and serum (r = 0.6396) and sputum ECP(r = 0.4683) in exacerbation. CONCLUSIONS: 1. In asthma exacerbation elevated sputum ECP concentration was observed. 2. In consequence of prednisone treatment the sputum ECP concentration was reduced. 3. Sputum ECP measurement is more accurate than serum ECP for monitoring the effectiveness of treatment. 4. Sputum ECP concentration is a sensitive parameter which discriminate asymptomatic patients with asthma from healthy subjects. PMID- 12884562 TI - [Analysis of pulmonary function tests and cellular composition of bronchoalveolar lavage fluid in patients with sarcoidosis]. AB - We investigated cellular composition of bronchoalveolar lavage fluid and changes in pulmonary function tests in patients with histopathologically proven sarcoidosis. We analyzed BAL, lung volumes, lung compliance, and diffusing capacity in 33 nonsmoking patients (18 males, 15 females, age 22-60 years, mean 40 years). In 39% of the patients we observed decreased static compliance and in 18% of patients reduction in diffusing capacity. The total cell count, percentage of lymphocytes and CD4/CD8 ratio and pulmonary function parameters in I, II and III radiological stages of sarcoidosis were similar. We found a moderate negative correlation between the carbon monoxide diffusing capacity (DLco% of pred.) and CD4/CD8 ratio (r = -0.40, p < 0.05). No correlation was observed between other pulmonary function indices and total cell count in BAL, percentage of lymphocytes and CD4/CD8 ratio. Our results suggest, that reduction in diffusing capacity may reflect an insensitivity of alveolitis, but correlation is moderate and it usefulness for making therapeutic decision is not clear. PMID- 12884563 TI - [Myeloperoxidase (MPO) as a marker of neutrophil influx into nasal mucosa after recombinant IL-8 challenge]. AB - Myeloperoxidase (MPO) has been proposed to mirror the degree of neutrophil activation, however its role as a marker of the participation of neutrophils in allergic inflammation is still unclear as the literature is controversial. The aim of this study was to evaluate the MPO levels and neutrophil influx in nasal lavage after recombinant Il-8 challenge. Eight patients suffering from seasonal allergic rhinitis, hypersensitive to grass pollens, with average age 30.1 +/- 2.67 were challenged both with Il-8 and diluent for Il-8 on a subsequent day, in two phases: before the pollen season (unprimed mucosa) and during the season (primed mucosa). Nasal lavage with saline were collected before, during Il-8 or placebo challenge and 30 minutes, 2 hours and 3 hours after the challenge. The number of neutrophils and MPO levels in the nasal fluid were determined. After the challenge with Il-8 of primed mucosa the number of neutrophils increased from 28250 cells/ml at the baseline to 28778, 251020 and 333660 at 0, 5, 2 and 3 hours respectively. There was a statistically significant difference between cells number after diluent and Il-8 challenge (p < 0.05, Wilcoxon rank-sum test). The number of neutrophils in the nasal lavage of primed mucosa after Il-8 challenge was significantly higher (p < 0.005) at all time points in comparison with the diluent and Il-8 challenges in the unprimed mucosa. There was no difference (p < 0.05) in MPO levels in the nasal lavage between Il-8 (mean 17.43 ng/ml +/- 10.98) and diluent challenge (20.98 ng/ml +/- 11.89) of unprimed mucosa. In the primed mucosa fluid we observed a peak of MPO level at 2 hours time point, however that was not significant as compared to diluent challenge (p = 0.465). We did not find the relationship between MPO levels and the neutrophils number in the lavage (rank Spearman factor, rs = 0.258, p = 0.42). Due to the lack of statistically confirmed relationship between MPO level and the number of neutrophils, MPO seems to be of little value as a marker of neutrophil influx into nasal mucosa. PMID- 12884564 TI - [Pulmonary mycobacterioses--frequency of occurrence, clinical spectrum and predisposing factors]. AB - Various types of non-tuberculous mycobacteria can be the aetiologic factors of chronic lung infections especially in patients with underlying chronic lung diseases. The aim of this study is to present the cases of pulmonary mycobacterioses observed in Institute of Tuberculosis and Lung Diseases in the years 1995-2001. There were 23 patients, 12 men and 11 women in the age between 35-77 years, mean 56 years. 16 out of 23 patients had underlying respiratory problems, mainly healed tuberculosis (7) and COPD (6). Two additional patients suffered from other diseases with potential immunosuppression (leukopenia). In 5 patients no disease other than mycobacteriosis was found, but they were chronic smokers. In 19 cases cough and expectoration of purulent sputum lasting from several months to several years was observed. In 5 patients onset of disease was acute or subacute with high fever. Eight patients had haemoptysis. In chest X-ray pathological lesions including (18 cases) lung cirrhosis (10) and cavities (15) were found. In 4 cases disseminated bronchiectases with small nodules were the main radiologic feature. Mycobacteriosis was caused by M. kansasii in 11 cases, by M. intracellularae in 6, by M. xenopi in 5 and by M. scrofulaceum in 1 case. PMID- 12884565 TI - [Preliminary results of prophylactic program of allergic diseases in children in Lodz district]. AB - Allergic diseases are one of the most important problems in medicine. As a consequence of increased frequency of allergic diseases, negative health, economical and social problems appear. To eliminate these consequences prophylactic programmes are created. In this paper preliminary results of Prophylactic Program of Allergic Diseases in Children in Lodz district in 2000 2001 years is presented. PMID- 12884566 TI - [Oxygen saturation at rest, on exercise and during sleep in COPD patients undergoing pulmonary rehabilitation program. Two years prospective controlled study]. AB - The role of pulmonary rehabilitation (PR) in COPD patients is well established. However, the PR programs includes physical exercise that may deteriorate chronic hypoxemia. The objective of the study was to evaluate impact of a long term PR program on oxygen saturation at rest, on exercise and during sleep in COPD patients no receiving supplemental oxygenation. A two--year controlled study involved 27 rehabilitated (PR-COPD patients (6 females) aged 32-76 years, mean 58.1 years on average and 19 control (C-COPD patients (3 females) aged 58-83 years, mean 68. COPD patients participating in PR program patients were younger, had higher value of FVC and FEV1 and better results of METs during symptom limited exercise test (ET, p = 0.007). They also had better results of transcutaneus oxygen saturation (SaO2) at rest (p = 0.02), a lower maximal decrease in SaO2 during ET(p = 0.003) and a higher value of medan SaO2 during (p = 0.02). Following PR program the number of METs achieved during ET increased in PR-COPD patients (p = 0.007) and did not change in C-COPD, p = NS. The SaO2 at rest did not change significantly in both groups. Maximal drop in SaO2 during ET decreased in PR-COPD patients (p = 0.02) and did not change in C-COPD(p = NS) group. Median SaO2 at sleep did not change in PR-COPD patients (p = NS) but it decreased in C-COPD patients (p = 0.02). The study indicates that long term PR does not deteriorate the resting diurnal hypoxaemia, it prevent COPD patients from time depending exacerbation of nocturnal hypoxaemia and can improve oxygen saturation on exertion. The results could be influenced by baseline differences between PR-COPD and C-COPD patients. PMID- 12884567 TI - [Prognostic value of some clinical, radiological, laboratory and functional parameters in sarcoidosis]. AB - Sarcoidosis is a granulomatous disease of unknown origin characterized by the trend to spontaneous remission in the great number of the patients. Some patients require treatment with corticosteroids, which have significant toxicity. The aim of this study was to assess the prognostic role of many different parameters in the patients with sarcoidosis. 162 sarcoidosis patients were introduced to prospective study: 22 patients were treated because of deterioration in lung function or serious ocular disease and 140 were observed without therapy for two years. We assessed the age, sex, symptoms, serum activity of angiotensin converting enzyme (SACE), hypercalcaemia, hypercalciuria, splenomegaly and HRCT findings at the time of diagnosis. We analyzed the frequency of spontaneous remission of sarcoidosis in the untreated patients. We investigated correlation between these parameters and remission. Statistical comparisons were made with chi-square test. We also applied the k nearest neighbor (k-NN) rule and the leave one out method adopted from the statistical pattern recognition theory. From many different parameters only acute symptoms (erythema nodosum, fever, arthritis) and serum activity of ACE might be helpful in predicting prognosis in the patients with stage I of disease. The patient's age at onset less than 36 years, the appearance of erythema nodosum and ground-glass opacities on HRCT scans portend an excellent prognosis in the patients with stage II of disease. PMID- 12884568 TI - [Wegener's granulomatosis--disease course and treatment response of 18 patients]. AB - Eighteen patients (12 female) with Wegener's granulomatosis (WG) treated from 1982-2001 were studied. Mean duration of patients' symptoms prior to diagnosis was 13.5 months, and mean age 41.2. Lung biopsy confirmed WG in 5 patients, nasal biopsy was made in 12, endobronchial in 3. In 11 of 14 patients cANCAs were found. Prednisone and 2 mg/kg/day doses of oral cyclophosphamide were used to treat 17 patients, prednisone was used alone for 1. Patients with severe symptoms were given i.v. 'pulse' methylprednisolone. Mean duration of treatment was 34.8 months and of follow-up, 71.9 months. A total of 4 patients died: 3 from complete renal failure. Four patients still undergo treatment, improving markedly. Complete remission was achieved in 6 patients. A total of 3 patients relapsed and are recently treated. In another one chronic renal failure developed. Three of them had moderate symptoms at diagnosis but were treated inadequately. IN CONCLUSION: manifestation of WG and severity of symptoms may be different among patients. ANCA is not universally present in all patients with WG. The absence of ANCA does not exclude WG diagnosis. Complete remission can be achieved in WG patients, including the severely ill, if adequate therapy is given. Moderate symptoms at presentation do not justify less intensive treatment. PMID- 12884569 TI - [Wegener's granulomatosis--a disease of many faces]. AB - We describe a case of Wegener's granulomatosis. The first symptoms included severe headache subsiding only after administration of dexamethasone. Despite a great number of diagnostic tests involving CT and MR of the head, the cause of the headache remained unknown. Because a chest x-ray revealed a nodule in the right lung, the patient was sent to our Clinic. Reevaluation of CT and MR pointed to a massive ethmoid sinusitis. 7-days' course of antibiotics and corticosteroids induced remission of the lung nodule. Several diagnoses were made: neoplasm, bacterial ethmoid sinusitis, trigeminal neuritis, thrombotic cavernous sinusitis and tuberculosis. Results of the ethmoid sinus biopsy together with a high c-ANCA concentration gave the correct diagnosis. PMID- 12884570 TI - [Tracheobronchopathia osteochondroplastica]. AB - Tracheobronchopathia osteochondroplastica (TO) is a rare disease of unknown etiology affecting mainly the trachea and large bronchi. It is characterized by the presence of multiple submucosal osseus and/or cartilaginous nodules. The authors report a case of 74-year-old woman in whom fiberoptic bronchoscopy, performed because of hemoptysis, revealed typical feature of TO. Besides the typical nodules protruding into the lumen of trachea and main bronchi, a small soft nodule in the larynx was found. On histological examination it was showed to be polyp with regions of inflammation and necrosis. The direct relation between such a laryngeal polyp and TO seems to be very unlikely. PMID- 12884571 TI - [Review of quality of life assessment methods in patients with non-small cell lung cancer]. PMID- 12884572 TI - [Potential role of viruses in pathogenesis of idiopathic pulmonary fibrosis]. PMID- 12884573 TI - [Pulmonary obstruction in spirometry and what next? How to correctly perform and assess the reversibility test]. PMID- 12884574 TI - [National parasite control programmes at the crossroads]. PMID- 12884575 TI - [Schistosomiasis control in China: progress and challenges]. PMID- 12884576 TI - [A case report of infant amebic dysentery]. PMID- 12884577 TI - [Development of information technology and public health information systems]. PMID- 12884578 TI - [An imported case of falciparum malaria from Laos]. PMID- 12884579 TI - [Impact of a reservoir project on schistosomiasis transmission in lake region]. AB - OBJECTIVE: To determine the impact of a project of building dike for storing water on schistosomiasis transmission in Junshan Lake. METHODS: Junshan Lake in Jinxian County of Jiangxi Province was selected as survey pilot. Data on snail distribution and historical prevalence of schistosomiasis before building dike were collected. The water level inside and outside of the dike was recorded from 1995 to 2002, and the relationship between the water level and snail population at the inside of the dike was analyzed. Survey was made in a natural endemic village to confirm the current endemicity of schistosomiasis. RESULTS: In the selected area of Junshan lake, schistosomiasis was prevalent in 6 villages of 3 townships, with a snail area of 1,394,030 m2 (2,090 Chinese Mu). Snails distributed mainly in the marshland at an elevation of 16.6-17.2 m, and the average infection rate of schistosomiasis in the residents was 12.5% in 1958 before the dike project. In 1960, two years after the dike was built, no living snails were found on the marshland, and the infection rate of schistosomiasis in the residents reduced remarkably. Currently, no schistosomiasis cases were found in human being and cattle in the surveyed village. CONCLUSION: The reservoir project had helped the elimination of snails and interrupted schistosomiasis transmission. PMID- 12884580 TI - [Cloning and expression of specific antigen genes of Ancylostoma caninum]. AB - OBJECTIVE: To search for the gene encoding specific antigen of Ancylostoma caninum that induces host's protective immunity. METHODS: A lambda ZAP II cDNA library of A. caninum was screened with sera from dogs immunized subcutaneously with hookworm larvae(L3). After sequencing, insert gene (AcAg) from positive clones was ligated into PUC18 and PET28C. Recombinant pET28C plasmid was induced to expressed protein in the E. coli BL21. The characteristic of recombinant protein is analyzed by SDS-PAGE and Western blotting assay. RESULTS: Five positive clones were obtained, and proved to be the same. The insert gene (AcAg) in pET28C vector expressed a recombinant protein of about 43 kDa. Using Western blotting assay, this protein was recognized by the sera from dog immunized with Ancylostoma caninum third-stage infected larvae and used for screening library. CONCLUSION: The AcAg, which exhibits 35% homologous to Caenorhabditis elegans gene unc-89, is a novel specific antigen of A. caninum. Its ability to elicit the protective immunity and the probability of the recombinant protein as a vaccine need to be further evaluated. PMID- 12884581 TI - [Clinical observation on 32 cases of paragonimiasis]. PMID- 12884582 TI - [Prokaryotic expression and characterization of an antigenic gene of adult Trichinella spiralis]. AB - OBJECTIVE: To obtain a purified Ts87 gene expression product of adult Trichinella spiralis and identify its immunogenicity. METHODS: Ts87 cDNA was subcloned into PET-28a(+) expression vector. The transformed bacteria bearing PET-28a(+)/Ts87 plasmids were induced by IPTG for production of fusion proteins. The expression product purified with His-binding affinity chromatography and electro-elution assay was analyzed by SDS-PAGE, ELISA and Western blot, and was used to immunize the rabbits. RESULTS: PET-28a(+)/Ts87 transformed bacteria produced the desired fusion protein with a relative molecular weight of 40 kDa. The antisera with high titer were obtained by immunizing rabbit with Ts87 recombinant protein. Ts87 expression protein was detected as positive reaction with infected rabbit sera, infected swine sera and antisera against Ts87 by ELISA. Ts87 protein was also recognized by above-mentioned sera with Western blotting. However, Ts87 protein failed to react with the patient sera infected with Cysticercus cellulosae or Echinococcus granulosus, and rabbit sera infected with Schistosoma japonicum. CONCLUSION: A new Trichinella recombinant protein with specific antigenicity was obtained. PMID- 12884583 TI - [Emergency treatment of sustained epilepsy in cerebral cysticercosis]. PMID- 12884584 TI - [Identification of Taenia saginata by mtCO I in four areas of Yunnan and Guizhou provinces]. AB - OBJECTIVE: To identify the types of Taenia saginata isolated from Dali of Yunnan Province, and from Duyun and Congjiang of Guizhou Province. METHODS: Genomic DNA was isolated, and the mitochondrial cytochrome C oxidase subunit I (mtCO I) genes were amplified by polymerase chain reaction(PCR) and analyzed by PHYLIP software package. RESULTS: The mtCO I gene sequences of Lanping sample were identical to that of T. saginata asiatica known in Taiwan, and the samples obtained from Dali and Duyun showed the same mtCO I gene sequences, while the sample from Congjiang had the same mtCO I gene sequences as T. saginata. The homology between these two groups of gene sequences was 97.44%, while the homology of amino acid sequences reached to 99.16%. The constructed phylogenetic tree revealed that the relationship between T. saginata asiatica and T. saginata is closer, both are distant relative to T. solium and other species of cestodes. CONCLUSION: The Taenia prevalent in Lanping, Dali and Duyun is identified as T. saginata asiatica, while that isolated in Congjiang is the typical T. saginata. PMID- 12884585 TI - [A misdiagnosed case of paragonimiasis]. PMID- 12884586 TI - [Protective effect against Schistosoma japonicum of recombinant fusion protein SjGST-32 in mice]. AB - OBJECTIVE: To explore the optimal immune doses of recombinant antigen rSjGST-32, with QuilA as adjuvant. METHODS: BALB/c mice were immunized with doses of 50, 100 and 200 micrograms rSjGST-32/mouse plus 50 micrograms QuilA adjuvant. QuilA and PBS were used as controls. Levels of specific antibodies were detected by ELISA. The mice were challenged 4 weeks after last vaccination. Worms and eggs collected from the livers of mice were counted 45 days after challenge. RESULTS: As compared with the control groups, the worm reduction rate in the 50, 100 and 200 micrograms experiment groups was 38.1%, 47.8% and 48.8%, respectively. The reduction rate of liver eggs per gram (LEPG) was 39.1%, 53.5% and 53.6%, respectively, and the reduction rate of the liver eggs per female (LEPF) was 22.5%, 22.8% and 21.8%, respectively. The specific antibody titers in sera reached 1:51,200, 1:102,400 and 1:102,400, respectively before challenge. CONCLUSION: The results show that for vaccinating BALB/c mice, the optimal antigen dose is 100 micrograms of recombinant rSjGST-32 plus QuilA adjuvant. PMID- 12884587 TI - [Taxonomic status in DNA sequences of five species of genus Paragonimus]. AB - OBJECTIVE: To analyze the taxonomic status of Paragonimus hokuoensis, P. paishuihoensis, P. menglaensis, P. bangkokensis and P. xiangshanensis in Genus Paragonimus Braun, 1899. METHODS: DNA sequences were obtained from the ITS2 and CO I genes and phylogenetic trees were built from these. RESULTS: There are some DNA sequence differences among P. paishuihoensis, P. menglaensis, P. bangkokensis and P. xiangshanensis, but the differences are small. P. hokuoensis and P. skrjabini are similar in DNA sequence. CONCLUSION: P. paishuihoensis, P. menglaensis, P. bangkokensis and P. xiangshanensis are closer in the genetic relationship. Their taxonomic status lies between P. skrjabini and P. westermani. The genetic relationship between P. hokuoensis and P. skrjabini is very close. PMID- 12884588 TI - [Two cases with gum myiasis caused by Wohlfahrtia magnifica]. PMID- 12884589 TI - [Observation on the amount of oxygen consumption by Oncomelania hupensis under low temperature]. AB - OBJECTIVE: To observe hibernation phenomena of Oncomelanai hupensis and explore the way of inducing the hibernation in laboratory. METHODS: Snails, O. hupensis hupensis, were collected from marshland of Jiangsu. The snail hibernation was induced by the way of cultivation at a mimic natural environment in the laboratory with gradually changing temperature. The amount of oxygen consumed by snails was tested by iodine titration, and their hibernation was tested by pin puncture followed by warm water. RESULTS: There was no significant difference on the rate of snail hibernation when the temperature was reduced by 1 degree C per 24 hrs and by 1 degree C per 48 hrs. The hibernation rate increased with the decreasing temperature. There was a significant regression relationship between hibernation rate and temperature with R2 = 0.967 (F = 207.72, P < 0.01). The temperature for 50% snails at hibernation (ET50) was at 5.87 degrees C with 95% confidence limit of 5.32-6.23 degrees C. The amount of oxygen consumed by snails declined with reduced temperature, there was a significant regression relationship between oxygen consumption and temperature with R2 = 0.963 (F = 182.18, P < 0.01). A significant regression relationship was also shown between oxygen consumption and hibernation rate (R2 = 0.916, F = 75.88, P < 0.01). CONCLUSION: Snail hibernation can be induced by the way of gradually decreasing temperature, and pin puncture or warm water resuscitation can be used to determine the status of snail hibernation. PMID- 12884590 TI - [Malaria incidence and the importance of floating population in malaria control in Tengchong County 1997-2001]. PMID- 12884591 TI - [Cloning and characterization of mitochondrial NADH dehydrogenase gene of Cysticercus cellulosae]. AB - OBJECTIVE: To clone and characterize the NADH1 gene of Cysticercus cellulosae. METHODS: A cDNA library was constructed from Cysticercus cellulosae and was immunoscreened by using rabbit anti-Cysticercus cellulosae polyclonal antibody. The gene structure and its possible function were analyzed by comparing with sequences available in the GenBank, after the insert of positive clone was subcloned and the nucleotide sequence of the insert was determined by dideoxynucleotide chain termination method. RESULTS AND CONCLUSION: A cDNA clone (named TS5) with a length of 1,082 bp was isolated. The 5' terminal of cloned gene contained one open reading frame of 1-578 bp encoding 192 amino acid residues of mitochondrial NADH dehydrogenase subunit 1 and the 3' terminal contained three kinds of tRNA genes. PMID- 12884592 TI - [Intranasal or intragastric vaccination of mice with recombinant Schistosoma japonicum ferritin induces immune protection against challenge infection]. AB - OBJECTIVE: To test the immune protection against challenge infection in mice vaccinated intranasally or intragastrically with recombinant Schistosoma japonicum (S.j) Ferritin (rSjFer). METHODS: Mice were divided into 8 groups each with 10 mice. They were immunized intragastically with rSjFer, CTB, rSjFer + CTB and intranasally with rSjFer, CTB and rSjFer + CTB respectively. PBS was used intragastically or intranasally as control groups. The mice were challenged with 40 +/- 1 S. j cercariae per mouse 2 wk after the third vaccinization. Forty-five days later, mice were killed and perfused, and the adult worms and eggs were counted. Serum and fecal samples were obtained before the first immunization and the challenge infection. IgA and IgG in sera and sIgA in feces were detected by ELISA. RESULTS: The worm reduction rate was 3.98%, 3.77%, 25.57% in the intragastric vaccination groups and 7.59%, 4.50%, 33.35% in the intranasal vaccination groups respectively. The egg reduction rate was 3.76%, 2.46%, 34.75% and 4.40%, 0.06%, 60.10% respectively. CONCLUSION: This study showed that a significant immune protection against Schistosoma japonicum infection was induced by mucosal (intranasal and intragastic) vaccination with rSjFer. PMID- 12884593 TI - [Two dimensional measurement of the mature and immature eggs of Schistosoma japonicum]. PMID- 12884594 TI - [Study on the level of specific IgG, IgG1 and IgE during anaphylactic shock in sheep induced by Echinococcus granulosus]. AB - OBJECTIVE: To investigate the change of specific antibodies IgG, IgG1 subclass and IgE in sheep infected with Echinococcus granulosus (E. g) during anaphylactic shock, and to observe antigen B reactivity against IgG antibody in E. g-infected sheep. METHODS: Antigen B and crude antigen were prepared with E. g cyst fluid (EgCF) from infected sheep. The enzyme-linked immunosorbent assay(ELISA) was used for detecting the level of specific IgG, IgG1 and IgE during anaphylactic shock in sheep induced by E. g. RESULTS: The level of specific IgG, IgG1 and IgE was significantly higher in the infected sheep after 6 months than that of the uninfected control group (P < 0.01). The IgE level decreased rapidly after anaphylactic shock induced, especially when the sheep was dying. Following an antigen challenge the sheep showed a general decrease in total IgG and IgG1 subclass. The total IgG showed a slight change at the beginning, followed by a decrease 1 h after challenge. The decrease of IgG1 subclass was more significant than the total IgG in 40 min after challenge injection. The positive rate was 91% for antigen B and 32% for crude antigen of EgCF against IgG antibody in E. g infected sheep. CONCLUSION: The specific IgE plays a major role in the anaphylactic shock, while IgG and IgG1 antibodies are also important. Antigen B derived from sheep E. g cyst fluid appears to be useful in serodiagnosis of and monitoring on the infection status in sheep. PMID- 12884595 TI - [Mechanisms of cytoadherence in severe cerebral falciparum malaria]. PMID- 12884596 TI - [Research progress in molecular biology of important proteins of Clonorchis sinensis]. PMID- 12884597 TI - [Application of microwave irradiating ELISA for serological detection of schistosomiasis japonica]. PMID- 12884598 TI - [Human myiasis in China: a review of 107 cases during 1995-2002]. PMID- 12884599 TI - [Structure predication and function analysis on the stage-specific serine proteinase in newborn larvae of Trichinella spiralis]. PMID- 12884600 TI - [Diagnosis of falciparum malaria by immunocapture lactate dehydrogenase activity assay]. PMID- 12884601 TI - [Note of Schistosma sinensium in Napo County, Guangxi]. PMID- 12884602 TI - [An outbreak of trichinosis in Milin County of Tibet Autonomous Region]. PMID- 12884603 TI - [Explosion of Culex pipiens pallens in winter season and its control]. PMID- 12884604 TI - [Filariasis control and surveillance in Yongjia County from 1956 to 1998]. PMID- 12884605 TI - [Malaria surveillance in areas with different mosquito vectors in Chongqing municipality]. PMID- 12884606 TI - [Malaria surveillance during 1987-2001 in Jiande City]. PMID- 12884607 TI - [Data filing and management during filariasis elimination]. PMID- 12884609 TI - [Current situation and trends of the journals on parasitology]. PMID- 12884608 TI - [Isolation of the infective stage larvae of Ascaris lumbricoides]. PMID- 12884611 TI - [Research on blood-sucking blackflies (Diptera: Simuliidae) in China]. PMID- 12884610 TI - [A case of human infection with Rhabditella axei in Jiangsu Province]. PMID- 12884612 TI - [Study on DNA sequences of Paragonimus skrjabini populations from five provinces in China]. AB - OBJECTIVE: To study differences among Paragonimus skrjabini populations from five provinces (Guangdong, Fujian, Yunnan, Hubei, Sichuan) and Paragonimus szechuanensis and to analyze the taxonomic status of P. heterotremus and P. veocularis in the Genus Paragonimus Braun, 1899. METHODS: DNA sequences were obtained from the ITS2 and CO1 genes and phylogenetic trees were built. RESULTS: Difference of the DNA sequences among P. skrjabini populations from five provinces were minor. P. veocularis and P. skrjabini were similar in DNA sequences. There were also some resemblances between P. miyazakii from Japan and the Fujian isolates of P. skrjabin. In addition, P. heterotremus was found to be closer to P. skrjabini and quite a distance from P. westermani in the phylogenetic tree. CONCLUSION: All studied populations can be regarded as different strains of P. skrjabini; P. szechuanensis is not a separate species, but possibly a geographical strain of P. skrjabini. Also found was that P. veocularis and P. miyazakii may be the synonyms of P. skrjabini. As a separate species, P. heterotremus was found to be closer to P. skrjabini and quite a distance from P. westermani in the genetic relationship. PMID- 12884613 TI - [Prokaryotic expression of gene encoding Schistosoma japonicum SjE16 and its potential application in immunodiagnosis]. AB - OBJECTIVE: To sub-clone and express the gene encoding Schistosoma japonicum calcium-binding protein (SjE16) and study its immunological response. METHODS: The specific primers were designed according to the expressed sequence tags (ESTs) sequence, which was used for amplification of the encoding sequence from the cDNA clone containing SjE16. The gene was subcloned into pGEX4T-1 plasmid and expressed. The rSjE16 was tested for its immunological response by ELISA. RESULTS: The gene encoding Schistosoma japonicum SjE16 was cloned and expressed successfully. The immunogenicity and diagnostic potential of rSjE16 were investigated. It was demonstrated by immunoassay in rabbits that the specificity and sensitivity of the test were 94.1% (16/17) and 88.2% (15/17), respectively, and the level of antibody titer of the untreated group reached a peak at 9-11 wk post infection and maintained high at least for 21 wk post infection, while the antibody level in the treated group rapidly decreased to pre-infection level in 11 wk after treatment. In human, the specificity of the test was 98.3% (57/58); the sensitivity of acute and chronic patient serum assay was 85.5% (53/62) and 70.2% (40/57), respectively. CONCLUSION: The recombinant protein of SjE16 (rSjE16) was acquired. It can be recognized by the sera from schistosomiasis patients, and the level of antibodies decreased quickly after treatment in experimental rabbits, which implicates the potential value for the evaluation of chemotherapy and detection of active infection. PMID- 12884614 TI - [Protective antibody response to recombinant fragments of Plasmodium falciparum apical membrane antigen 1]. AB - OBJECTIVE: To determine the role of putative apical membrane antigen (AMA)1 domains in inducing protective immunity and to provide basis for selection of vaccine applicable segments. METHODS: Encoding gene segments of AMA1 were amplified and cloned into pET prokaryotic expression vectors. Recombinant proteins were expressed and purified. Groups of BALB/c mice were immunized by using recombinant protein in Freund's adjuvant, and the IgG titer and specificity of the immune sera were analyzed by IFA and Western blotting. Efficiency of the immune sera in inhibiting Plasmodium falciparum in vitro growth was evaluated. RESULTS: Recombinant AMA1 fragments including the entire ectodomain E and subdomain I + II, I, II and III were successfully expressed and purified. Different levels of antibody were induced in mice by individual proteins and all the immune sera recognized native antigen in the parasites. Sera from protein E and I + II immunized mice inhibited the growth of parasites. CONCLUSION: The integrality of the ectodomain of AMA1 determines the conformation of the protective antibody epitopes, and these protective epitopes distribute mainly in the subdomain I. PMID- 12884615 TI - [A case of cerebral sparganosis]. PMID- 12884616 TI - [Study on the relationship between intracellular free calcium and melanization in oocysts of Plasmodium yoelii]. AB - OBJECTIVE: To study the change of intracellular free Ca2+ in the oocyst when it melanized and to find out the relationship between the melanized oocyst and its intracellular level of free Ca2+ in a Plasmodium-refractory strain of Anopheles dirus. METHODS: The distribution and experimental condition of the intracellular free Ca2+ in oocyst of Plasmodium yoelii was measured with Ca2+ sensitive dye Fluo-3/AM and Pluronic F-127 under confocal laser scanning microscope (CLSM) at different time. RESULTS: The best load condition was that the oocysts were incubated in 3 mumol/ml Fluo-3/AM adding 1 microliter/ml 25% Pluronic F-127 for 60 min at 37 degrees C. Fluorescent imaging of oocysts was affected by an increase or decrease of the concentration of Fluo-3/Am and incubation time. The distribution of intracellular free Ca2+ was heterogeneous in the oocysts. The mean value of Ca2+ in the mature oocysts was (137.15 +/- 7.02) nmol/L (X +/- S) but was (18.44 +/- 1.75) nmol/L in melanized oocysts with Ca2+ sedimentation in the wall of oocyst. CONCLUSION: The results suggest that the level of the intracellular free Ca2+ in oocyst decreased and excreted during its melanization in a Plasmodium-refractory anopheline mosquito species. PMID- 12884617 TI - [Immunological screening of Toxoplasma tachyzoite cDNA expression libraries with serum from infected rats]. AB - OBJECTIVE: To screen and identify the potential candidates for the development of toxoplasmosis vaccine. METHODS: Rats were infected with Toxoplasma gondii (T. gondii) RH strain and their sera were used as a probe to screen T. gondii tachyzoite cDNA expression libraries. The positive clones were analyzed by PCR amplification and DNA sequencing. RESULTS: Thirteen positive clones were obtained from about 4 x 10(5) phage plagues after three rounds of screening. The size of the inserts ranged from 0.45 kb to 2.4 kb. A BLAST search of all available sequence databases using the partial sequences from four positive clones (L1, L2, L4, L5) showed that the sequence of L2 clone was identical with T. gondii P24 major antigen gene (TgP24). Clone L4 had a high homology with Saccharum officinarum pyruvate orthophosphate dikinase. There is no significant hit of any sequences to clone L1, suggesting that L1 could be a novel gene(GenBank accession number AY180109), named T.g-R1, which encodes a non-transmembrane protein with 134 amino acid open reading frame. PROSCAN analysis of the T.g-R1 amino acid sequence showed that this gene product contains two protein kinase C phosphorylation site, two casein kinase II phosphorylation site, one N myristoylation site and one microbodies C-terminal targeting signal. Clone L5 was a small partial fragment. CONCLUSION: The identification of positive clones provides a possible way for the development of toxoplasmosis vaccine. PMID- 12884618 TI - [The correlation between DDVP resistance of Culex pipiens pallens and esterase activity]. AB - OBJECTIVE: To detect the resistance index and esterase activity of each generation of DDVP-resistant Culex mosquitoes and analyze the relationship between insecticide resistance and esterase. METHODS: WHO bioassay and micro plate measurement were used for the detection. RESULTS: The resistance index increased to 12.17 after 43 generations' insecticide selection compared to 1.00 as sensitive isolate. The nonspecific esterase(NSE) activity of the mosquitoes became strengthened with the extension of the generations, and the individual frequency of those with OD values no less than 0.9 increased gradually, consistent basically to the bioassay. The AChE average inhibition rate decreased with the extended generation and increased resistance, and the individual frequency of those with inhibition rate less than 30% became strengthened with the extension of generations, showing a positive correlation. CONCLUSION: The activity of NSE and AChE shows a correlation with DDVP resistance. PMID- 12884619 TI - [Construction and expression of the recombinant plasmid pcDNA3.1/Ts87 of Trichinella spiralis]. AB - OBJECTIVE: To construct and express DNA plasmid of Trichinella spiralis. METHODS: The BamHI and HindIII enzyme sites and KOZAK sequences were introduced at both ends of Ts87 gene by PCR. Ts87 gene was ligated into pcDNA3.1(+) vector with T4 ligase. The recombinant plasmid pcDNA3.1/Ts87 and plasmid pcDNA3.1 were transfected into an eukaryotic cell line COS7 through Lipofectamine, respectively. The BALB/c mice were immunized with the purified plasmid DNA pcDNA3.1/Ts87 through two routes: intramuscular injection and gene-gun injection. RESULTS AND CONCLUSION: The pcDNA3.1/Ts87 was expressed both in COS7 and in the BALB/c mice. PMID- 12884620 TI - [Investigation on the impact of imported cases on filariasis elimination program in Shandong Province]. AB - OBJECTIVE: To study the impact of imported filariasis cases on the elimination program in different areas of Shandong Province. METHODS: Dezhou was selected as former low endemic area and Yicheng as former high endemic area. Blood examination was carried out for both mobile population and local people for microfilariae(Mf). Mosquitoes were caught in field and dissected to count the ratio of those having laid eggs and the natural filarial infection rate. Mosquitoes reared at different temperatures were fed with Mf-positive blood and dissected after certain time period to observe the development of the larvae. The vectorial capacity and case transmission quantity were calculated and compared with those from different areas. RESULTS: The Mf positive rate of inflow population was 3.18% in average. No case was detected from 9,411 local residents after blood exam in Dezhou while 2 out of 692 local residents were found Mf positive in Yicheng. Mosquitoes natural infection rate was 3.81% but no third stage larva was found. The shortest time period needed for the larva to develop into an infective stage was 16 days in Dezhou and 11 days in Yicheng. The time period from blood meal to egg-laying on average was 4.95 days in Dezhou and 4.33 days in Yicheng. The ratio of vectorial capacity and case transmission quantity was 1:4.41 and 1:5.82 respectively in Dezhou and Yicheng. CONCLUSION: Filarial transmission seems unlikely in Dezhou for its low vectorial capacity and low transmission quantity resulted from low and evidently fluctuating temperature in the north. A low level filarial transmission may be possible in former high endemic area such as Yicheng if there are as many imported cases as in Dezhou. PMID- 12884621 TI - [A malaria case with more than one ring form of Plasmodium vivax within an erythrocyte]. PMID- 12884622 TI - [Determination of free thiols in the chimeric protein PfCP-2.9 of Plasmodium falciparum]. AB - OBJECTIVE: To determine the free thiols in the chimeric protein PfCP-2.9 of Plasmodium falciparum expressed by Pichia pastoris. METHODS: Two experiments of reverse phase HPLC and Ellman's reaction were applied to the PfCP-2.9 for the determination of its free thiols. For RP-HPLC analysis, three kinds of samples were tested: PfCP-2.9, dithiothreitol-reduced PfCP-2.9 and indoacetic acid alkylated PfCP-2.9. RESULTS: Both experiments showed that there were no any free thiols present in the PfCP-2.9. CONCLUSION: The disulfide bonds between cysteine residues of PfCP-2.9 were formed completely. PMID- 12884623 TI - [Isoenzyme analysis on different isolates of Trichomonas vaginalis]. AB - OBJECTIVE: To study the biological types on the seven isolates of Trichomonas vaginalis from Beijing, Hebei-Tangshan, Hebei-Chengde and Jiangxi-Jiujiang in the mainland of China. METHODS: The samples were analyzed by PAGE, isoenzyme stain and cluster analysis. RESULTS: The isoenzyme systems used in the study included MDH, LDH, G-6-PD, PGI and PGM. No difference in the isoenzyme patterns of G-6-PD and PGI was found among the seven isolates. The MDH and LDH patterns of Beijing 1, Beijing 2, Jiujiang 3 strains were identical, while they were distinguishable from those of Chengde, Tangshan, Jiujiang 1, Jiujiang 2 isolates. The PGM pattern of Beijing 1 and Beijing 2 isolates were same but was different from that of the remainders. Gene tree was constructed according to the isoenzyme profiles. The results showed that there are differences in the patterns of the five isoenzymes between the isolates of Beijing 1, Beijing 2, Jiujiang 3 and other four isolates, and Jiujiang 3 was different from Beijing 1, Beijing 2 slightly. CONCLUSION: It seems reasonable to assume that there are at least three different biological types of Trichomonas vaginalis in China. PMID- 12884624 TI - [An investigation on the citation of Chinese periodicals in Chinese Journal of Parasitology and Parasitic Diseases]. PMID- 12884625 TI - [Construction of the plant expression vectors containing the multiepitope gene of Toxoplasma gondii]. AB - OBJECTIVE: To construct the plant expression vectors containing the multiepitope gene of Toxoplasma gondii (TGMG). METHODS: 1. TGMG was subcloned into pBAC55 vector to construct the intermediate plasmid pB35MG. The E35S/TGMG/NOS3' fragment was cleaved from pB35MG and ligated into the plant binary vector pCAMBIA2300 to construct the plant expression vector pC35MG. 2. Tomato fruit-specific E81. 1 promoter was introduced to pB35MG to construct pB35E1MG vector. The E35SE81. 1/TGMG/NOS3' fragment was subcloned into pCAMBIA2300 to construct the plant expression vector pC35E1MG. 3. Tomato fruit-specific E82.2 promoter was inserted to pB35MG to construct pBE2MG vector. The E82.2/TGMG/NOS3' fragment was subcloned into pCAMBIA2300 to construct the plant expression vector pCE2MG. The insert gene TGMG in the vectors pB35MG, pC35E1MG and pCE2MG were confirmed by sequencing. 4. pC35MG, pC35E1MG and pCE2MG were introduced into Agrobacterium tumefaciens strain LBA4404 competent cell. RESULTS: Digestion with restriction enzymes proved that all recombinant vectors had the inserts with expected length of the target fragments. And the sequencing results were confirmed correct. CONCLUSION: The TGMG intermediate vectors pB35MG, pB35E1MG and pBE2MG and the plant expression vectors pC35MG, pC35E1MG and pCE2MG were constructed successfully, and the three plant expression vectors were introduced into Agrobacterium tumefaciens. PMID- 12884626 TI - [Epidemiological study on group infection of Angiostrongylus cantonensis in Changle City]. AB - OBJECTIVE: To make etiological and epidemiological investigation on the infection of Angiostrongylus cantonensis in 8 pupils in Changle City. METHODS: 1. CSF of patients was examined with the conventional method to detect pathogens and eosinophiles. 2. The fecal samples of wild rodents were collected from the spot and examined microscopically to discover the first stage larvae of A. cantonensis. 3. Snails (Pila gigas) were collected in the spot. The smashed head tissue was examined for the third stage larvae of A. cantonensis. 4. The patient's clinical symptoms and physical signs were recorded with an emphasis on central nervous system. RESULTS: 1. Two larvae of the third stage of A. cantonensis were found in CSF of one patient. Eosinophiles occupied 68% of the cell number in average (ranged from 47% to 83%) in CSF of the 8 patients. 2. The infection rate of the first stage larvae of A. cantonensis was 39.3% (44/112) in feces of the rodents. 3. The infection rate of the third stage larvae of A. cantonensis was 40.0% (82/205) in the snails. 4. Major clinical manifestations in the 8 patients included: severe headache(8/8), dizziness(8/8), nausea(8/8), vomiting(8/8), lethargy(7/8), lower limb hypodynamia(7/8). CONCLUSION: The confirmation of severe infection of A. cantonensis in 8 child patients demonstrated that a natural nidus of angiostrongyliasis is present in Chengle City. PMID- 12884627 TI - [A misdiagnosed case of paragonimiasis]. PMID- 12884628 TI - [Clinical observation on the efficacy of ivermectin in the treatment of intestinal nematode infections]. AB - OBJECTIVE: To observe the efficacy and side effects of ivermectin in the treatment of intestinal nematode infections. METHODS: A single dose of ivermectin 0.1, 0.2, 0.2 and 0.2 mg/kg was orally administered to cases with infection of Ascaris, hookworm, Trichuris and Enterobius respectively. A single dose of albendazole 400 mg was used as control. RESULTS: The egg negative conversion rates of ivermectin and albendazole were both 100% (34/34) for Ascaris infection, 17.6% (6/34) and 76.5% (26/34) respectively for hookworm infection, 67.6% (23/34) and 47.1% (16/34) respectively for Trichuris infection, 58.8% (20/34) and 100% (34/34) respectively for Enterobius infection. The worm discharge reached a peak in 1-2 days after treatment. The side effect of ivermectin was mild and transient showing no adverse effect on blood picture, liver function, renal function or ECG. CONCLUSION: Ivermectin shows similar effect on Ascaris with albendazole, better effect on Trichuris and poorer effect on hookworm and Enterobius than albendazole. PMID- 12884629 TI - [Study on morphology of Blastocystis hominis in culture and from diarrhea patients]. AB - OBJECTIVE: To study morphology of different stage Blastocystis hominis (B. h) for establishing a base in the research of life cycle and pathogenicity of B. h and providing information for clinical laboratory. METHODS: B. h from diarrheal patients was continuously cultured in LES medium, and morphology of B. h was studied with iodine and iron hematoxylin staining under light microscope. RESULTS: The vacuolar, granular, amoeboid and cyst forms of B. h and transformation among the forms were observed microscopically. CONCLUSION: Among different forms, the vacuolar and granular forms were often seen clinically and the vacuolar form can transform to cysts. PMID- 12884630 TI - [The impact of sandflies and leishmaniasis research on the development of western China]. PMID- 12884631 TI - [HE staining of uterus cells of Ascaris suum by egg albumin-glycerine smear technique]. PMID- 12884632 TI - [Ten years' control of Enterobius vermicularis infection in children in Zhejiang Province]. PMID- 12884633 TI - [Seguence analysis of the ITS2 gene of a large metacercaria from Sinopotamon chekiangense]. PMID- 12884634 TI - [The prevalence and control of schistosomiasis in marshland of Tongling County]. PMID- 12884635 TI - [An investigation on intestinal helminth infections among residents of Harbin City]. PMID- 12884636 TI - [Effect of chemotherapeutic regimens on soil-transmitted nematode infections in areas with low endemicity]. PMID- 12884637 TI - The operative outcome in children with supravalvular aortic stenosis. AB - The authors present their experience in surgical treatment of children with supravalvular aortic stenosis (SAS). A retrospective analysis was carried out on 20 patients operated in the years 1980-1999. The group included 12 boys and 8 girls aged 0.9 to 14.5 years (mean 7.8 +/- 3.4 years). The diagnosis of Williams syndrome in 6 children (30%) was based on the typical face. The mean systolic pressure gradient across the SAS was 84.3 +/- 34.5 mmHg (range from 41 to 168 mmHg). Intraoperative studies allowed to determine that 16 (80%) children had the hourglass type, 2 (10%) tubular type and 2 (10%)--fibromuscular membrane type of stenosis. Sixteen (80%) children had anomalies of the aortic valve and/or coronary arteries, and in 13 (56%) children associated heart or great vessels anomalies were noted. All the patients were operated on using cardiopulmonary bypass and hypothermia. The associated anomalies were corrected simultaneously. There were no early and late deaths. Because of restenosis one child was reoperated. In the follow up the mean systolic pressure gradient between the left ventricle and ascending aorta revealed by echocardiography in 19 (95%) children to be 8 +/- 2 mmHg (range from 0 to 28 mmHg). Eighteen children are now in NYHA class I (New York Heart Association) and develop normally. Supravalvular aortic stenosis is a rare complex anomaly, which, because of life-threatening complications and low operative risk, should be fully repaired as soon as possible. PMID- 12884638 TI - [Treatment of osteoporosis and osteopenia in children--own experience]. AB - Osteoporosis and osteopenia diagnosed in developmental age require special treatment with careful consideration of indications for particular drugs, their dosage and monitoring of treatment. The classical method, similarly as in adults, is administration of calcium and vitamin D, physical rehabilitation, treatment of fractures and pain release. The aim of the study was to evaluate the effect of combined therapy of decreased bone mineralization in 45 children (28 boys and 17 girls) aged 6.5 to 18 years. In 15 of them secondary osteoporosis (13/15) or osteopenia (2/15) were diagnosed, and in 30 cases the disorders were primary 16/30 and 14/30, respectively. The patients were treated from 6 months to 4 years. All patients received calcium and vitamin D preparations. In the majority of treated children calcium-rich diet and physical rehabilitation were applied, adjusted to the advancement of the disease. In 6 cases treatment with bisphosphonates was given. In our study we also present results and observations from calcitonin treatment of 35 children aged 6 to 18 years. This treatment was applied during the last 5 years in various periods--in 23 children with secondary and in 12 with primary osteoporosis. The therapy of osteoporosis and osteopenia in developmental age should always be individually assessed to disease advancement, symptoms, concomitant illnesses, age and possibilities of long-term treatment. The classical treatment includes appropriate intake of calcium, vitamin D and other vitamins and minerals (pharmacological preparations, diet), physical rehabilitation and pain release. On the basis of our observations it appears that antiresorptive drugs such as calcitonin and bisphosphonates may be used in treatment of osteoporosis in developmental age. Evaluation of treatment efficacy should involve clinical improvement and results of additional examinations, especially densitometry. Among biochemical tests, bone resorption markers (Pyr, Dpyr, CrossLaps) appeared to be the most useful; treatment of bone fractures should be evaluated by X-ray examinations. PMID- 12884639 TI - [Selected new atherosclerosis risk factors and markers of fibrinolysis in children and adolescents with obesity, hypertension and diabetes]. AB - In last years it has been proved that atherosclerosis risk factors are present in children and adolescents, and that already in their young age they are connected with anatomic, atherosclerosis changes in vessels. The aim of the study was to evaluate levels of selected new atherosclerosis risk factors (Lp(a), apo A-I, apo B, homocysteine, fibrinogen) and markers of fibrinolysis (t-PA and PAI-1) in children and adolescents with traditional risk factors (obesity, hypertension, diabetes). MATERIALS AND METHODS: The study group consisted of 285 children and adolescents aged 14.3 yrs. Children were divided according to their main disease into groups: children with obesity (n = 49), children with obesity and coexisting hypertension (n = 56), children with hypertension (n = 58) and children with diabetes (n = 122). Control group consisted of 79 healthy children and adolescents aged 13.3 yrs. Lp(a), apo A-I and apo B levels were estimated by use of immunoturbidimetric methods, total homocysteine, fibrinogen, t-PA and PAI-1 were estimated by use of immunoenzymatic methods. RESULTS: Lp(a) level in the total study group was 0.30 g/L and was over twice higher than in the control group -0.14 g/L. Apo A-I level was significantly lower in obese children (1.27 g/L) and in the group with obesity and coexisting hypertension (1.25 g/L) vs 1.35 g/L in controls. Apo B level was significantly higher in the total study group (0.86 g/L) and in groups with obesity, obesity and coexisting hypertension and diabetic children vs 0.73 g/L in controls. Hcy was higher in the group with obesity and coexisting hypertension (8 mumol/L) and in the group with hypertension (9.4 mumol/L) vs 6.2 mumol/L in control group. FB level was higher in the total study group (2.76 g/L) and in groups of obese children (3.18 g/L) and obesity coexisting with hypertension (3.22 g/L) vs 2.52 g/L in controls. Significantly higher t-PA level was found in the obese group (9 micrograms/L) and obesity with hypertension group (9.7 micrograms/L) vs 7.3 mg/L in controls, and PAI-1 level was significantly higher in total study group (62.3 micrograms/L) and groups of obese children (73.8 micrograms/L), obese and hypertensive (78 micrograms/L) and hypertensive (73 micrograms/L) vs 42.4 micrograms/L in control group. 28% of study children had positive family history of cardiovascular diseases. CONCLUSION: 1. Young patients with obesity, hypertension or obesity present significant lipid metabolism disturbances, regarding mainly total cholesterol, LDL, triglycerides, and Lp(a) and apo B levels. Unfavourable lipid profile is characteristic mainly in children with obesity and coexisting hypertension. 2. Elevated homocysteine level is found in children with hypertension. 3. Elevated fibrinogen level and diminished fibrinolytic activity characterises obese children. 4. Children and adolescents with obesity, hypertension or diabetes often come from families with positive family history of cardiovascular diseases and other atherosclerosis risk factors. PMID- 12884640 TI - [Serum levels of interleukin-10 in patients with ovarian carcinoma in response to chemotherapy]. AB - The levels of IL-10 and CA-125 were measured in 48 patients with ovarian carcinoma in different histological types and stages of clinical advance. The measurements were performed before surgical procedure and after chemotherapy. Pretreatment levels of IL-10 and CA-125 were significantly increased in carcinoma patients relatively to the control group. The frequency of higher levels and absolute value of IL-10 and CA-125 increased with the stage of disease. They decreased after chemotherapy in patients responding, while they remained elevated in non-responding to the treatment. The presents study shows, that IL-10 especially in the combination with CA-125 may be useful in the diagnosis and the monitoring of therapy results in patients with ovarian carcinoma. PMID- 12884641 TI - [Serum lipofuscin level after renal transplantation]. AB - BACKGROUND: Lipofuscin is believed to be the last product of lipid peroxidation. Elevated serum lipofuscin in chronic renal insufficiency and in multiple organ failure after major surgery has been detected. AIM: To compare serum lipofuscin level after renal transplantation with hemodialysed patients and healthy adults. MATERIAL AND METHODS: Lipofuscin has been determined according to Tsuchida with Roumen's modification and expressed in arbitrary units (u.), in serum of 56 healthy adults, 11 renal transplant recipients and nine hemodialysed patients. Blood was collected before hemodialysis in uremic patients, or within 24 h after renal transplantation. RESULTS: Serum lipofuscin in hemodialysed patients and renal recipients was significantly higher about 80% and 180% respectively, comparing to healthy adults. CONCLUSION: Elevated serum lipofuscin levels after renal transplantations show high oxidative stress due to several mechanisms of lipid peroxidation in this group of patients. PMID- 12884642 TI - [Torasemide (Trifas) in clinical practice--own experience]. AB - Torasemide (Trifas, Berlin Chemie/Menarini) is a loop diuretic and its site of action in the nephron is the afferent section of Henle's loop. Associated with this is its increased effectiveness and reduction of side effects in comparison with other loop diuretics like furosemide. Torasemide acts proportionately to the administered dose, which allows to asses its diuretic effect. It has been used in our Clinic in patients not responding to previous diuretic treatment. The diuretic effect of torasemide has been satisfactory, with no side effects. However, the trial group has not been numerous and was non-homogeneous in regard to renal failure and other disorders. PMID- 12884643 TI - [Effects of statins on coronary artery disease. Review of the recent publications]. AB - This review describes the role of statins in the primary and secondary prevention of coronary artery disease and the clinical benefit of therapy with statins in different forms of coronary artery disease (stable, unstable and myocardial infarction). Presented clinical trials have provided data that statins not only reduce plasma total cholesterol and LDL-cholesterol levels but also reduce major coronary event rate and all-cause mortality. Treatment with statins is usually well tolerated. The mean total cost of care to reach LDL-cholesterol target is significantly lower with atorvastatin than with the other statins. PMID- 12884644 TI - [Hemodynamic consequences of atrial fibrillation]. AB - Beside supraventricular extra beats, atrial fibrillation is the most common cardiac arrhythmia. Atrial fibrillation according to Douglas P. Zipes definition is an arrhythmia characterised by complete asynchronous atrial depolarisation, without following hemodynamic effective contraction. Atrial electric activity can be detected during electrocardiography as irregular oscillation of the isoelectric line of low and variable amplitude 350-600/min waves. Atrial fibrillation previously considered as a benign arrhythmia is associated with severe clinical complications as hemodynamic disturbances and closely connected with it cerebral embolization. In the present study we focused on left ventricle function during atrial fibrillation. We evaluate a deterious effect of high, irregular heart rate which stimulates left ventricle systolic dysfunction, so called "tachycardiomyopathy". Deterioration of left ventricle systolic function usually coexists with diastolic impairment, which is difficult to treat even if sinus rhythm is restored. We also analysed the pathophysiologic mechanism of left ventricle impairment during atrial fibrillation at cellular level. In second part of this paper we discussed dependence between size and function of left atrium and its appendage and atrial fibrillation induction. One must take into account of vicious circle mechanism induction during atrial fibrillation, which promotes arrhythmia stabilization, as well as after sinus rhythm restoration--may provoke recurrence of arrhythmia. Intracellular changes followed by electrophysiological alteration are regarded as causes of the mentioned mechanism. Accordingly to the newest studies, gene expression plays a role in arrhythmia induction and stabilization. This means that the problem is very complicated and also gives hope to effective treatment of atrial fibrillation including gene modification. PMID- 12884645 TI - [Leptin--the metabolic signal from adipose tissue]. AB - Leptin is a hormone that is released mainly by adipocytes. One of the roles of leptin in animals is regulation of the energy balance by decreasing food intake and increasing energy expenditure. Beyond this function, it influences sexual and reproductive system development. Much evidence suggests that leptin plays the same roles in human beings. It is also believed that this hormone takes part in the regulation of hematopoietic, endocrine (other than reproductive) and sympathetic system functioning, and is involved in pathogenesis of arterial hypertension and diabetes. This brief overview focuses on the history of leptin discovery, regulation of circulating leptin levels, and physiological role (scientifically proved and being under research). The use of recombinant leptin in patients with obesity is mentioned. PMID- 12884646 TI - [Resistine--a new hormone secreted by adipose tissue (adipose tissue in insulin resistance)]. AB - Obesity is an important risk factor for the development of insulin resistance and type 2 diabetes. The molecular mechanism linking obesity to insulin resistance is, however, unclear. Recently, a new circulating hormone resistine, which is expressed in adipose tissue, has been identified. Resistine has been shown to antagonize insulin action. Resistine levels are increased in diet-induced obesity as well as in genetic models of obesity and insulin resistance. Furthermore, resistine gene expression is markedly downregulated by treatment with anti diabetic drugs called thiazolidinediones, that improve target-tissue sensitivity to insulin. It has been found that in human abdominal adipose tissue, which is thought to be a main risk factor for insulin resistance, amount of resistine mRNA is higher than in other fat depots. Resistine, therefore, may play a role in the pathogenesis of obesity-related insulin resistance. PMID- 12884647 TI - [Cystic lesions--diagnosis and conservative treatment]. AB - Effective treatment of cystic lesions that we can apply in the outpatient clinic is the target for seeking a new solution in finding treatment that will produce a better percentage of recovered patients. At this moment non-surgical treatment of cystic lesions is compare with traditional methods of treatment i.e. cyst content aspiration and injection of steroids, with methods which lead to obliteration of cyst lumen and in this way closing the space being the reservoir for the cyst content. Local application of hialuronidase, aspiration, and then steroid injection is gaining more and more attention of clinicians. We also pay much more attention to local fibrinogen injection as a safe and effective method of treatment, which can be useful in the treatment of cystic lesions and topical treatment of bursitis. PMID- 12884648 TI - [Coronary artery stenting in the treatment of 5-fluorouracil-induced unstable angina]. AB - Cardiotoxicity is a rare but very serious side effect of 5-fluorouracil (5-FU) treatment. Many theories have been suggested to explain the mechanism of this problem. Most commonly, coronary artery spasm is considered responsible in cardiovascular toxicity. Ischemic symptoms and signs related to 5-FU therapy are observed during the late phase of the administration of the drug. The close and careful monitoring of all patients, especially the ones with pre-existent coronary artery disease, during 5-FU infusion is mandatory. Because there is not a single and effective modality of prevention of 5-FU cardiotoxicity, the patients should be selected carefully for its administration, 5-FU infusion should be stopped as soon as symptoms are encountered. A case of a 58-year-old woman treated with 5-FU due to colon cancer is described. In the late phase of 5 FU administration, the patient developed anginal pain with transitional ST segment elevation in ECG. Patient, after coronary angiography, was successfully treated with percutaneous coronary intervention. Interventional methods, most of all stent implantation, seem to be the best treatment of 5-FU-related acute coronary syndromes. PMID- 12884649 TI - [Pacemaking in Cracow in years 1966-1980]. AB - Permanent pacemaking, a new and effective bradycardia--treatment method, has appeared in the sixties. In Cracow the first emergency temporary stimulation by means of transvenously inserted intracardiac electrode was performed in 1966. Permanent technique of artificial pacing was introduced soon after. The number of pacemaker implantations grew successfully, obtaining 750 of the end of the seventies when the Institute of Cardiology of Nicolaus Copernicus Medical Academy was established. Assortment of implantable devices also became wider, including such rare constructions like nuclear pacemakers and an inductive coupled pacemaker. PMID- 12884650 TI - [Dr. Julian Wisz (1919-2000)--radiologist, educator and social worker from Rzeszow]. AB - This article is devoted to Dr Julian Wisz M.D., one of the Nestors of radiology. Already the first years after graduation he actively worked as a doctor who was responsible for the radiology service in Rzeszow and one of the faunders ambulance service. In the history of medicine in Rzeszow, Dr. J.Wisz was known as a social worker who participated actively in the works of the Polish Red Cross and also helped to organize the so called White Sundays, during which teams of doctors visited remote places to help people with medical problems. J. Wisz was also appreciated as the author of many talks and lectures concerning tuberculosis prophylaxis and alcoholism treatment. Among many activities he was also a lecturer at the Medical Vocational School and initiated mammography, which has been carried out in the oncological ward in the Provincial Hospital in Rzeszow since 1982. He was awarded many orders for his pedagogical and vocational work. Among them he received the Chivalrous Cross of the Order of Revival of Poland and "Pro Gloria Medici". PMID- 12884651 TI - [One-stage approach to the anterior and posterior vertebral column in operative treatment of advanced traumatic and neoplastic changes of the thoraco-lumbar spine]. AB - There were presented indications and operative technique of one-stage approach to anterior and posterior part of the thoraco-lumbar spine. Presented technique gave chance to control antero-lateral and posterior aspects of the spine. There were 2 patients with delayed compressions of the vertebral bodies in lumbar part with severe kyphotic deformity. Those deformities were the cause of progression of neural symptoms in a few months after injury. Another 4 patients had the spinal tumours. In one case it was giant-cell tumour which destroyed body and the arch of Th12. In two patients metastases of renal cancer infiltrated L2 and L3. The last one had breast cancer metastasis into Th12. In all patients it was necessary to control anterior and posterior columns of the spine to restore its axis and the shape of spinal canal. PMID- 12884652 TI - [Surgical treatment of neoplasm metastasis to the spine]. AB - The paper presents the treatment mode of metastasis to the spine. Between 2000 and 2002 5 cases (3 men and 2 women) with metastasis to the lumbar spine were treated surgically. The metastasis derived from primary foci in the prostate (3 cases), in the breast (1 case) and in the kidney (1 case). The average age of the patients was 65 years (ranging from 53 to 74 years). In all cases a vertebrectomy was performed through an anterior approach. Titanium baskets filled with bone cement were implemented. The spine was stabilized using anterior Kaneda instrumentation (3 cases) and the posterior Omega instrumentation (2 cases). This mode of treatment yielded good results. Pain relief was achieved by decompressing the neural elements, leading to better quality of life among the operated patients. PMID- 12884653 TI - [Treatment of displaced supracondylar fractures of the humerus in children at the orthopaedic ward of the Children's Hospital in Kielce]. AB - Ninety five children in age from 4 to 12 years (61 boys and 34 girls) with displaced supracondylar distal humerus fractures were treated at the orthopaedic ward of The Children's Hospital in Kielce between I 2000-XII 2001. The method of choice was closed reduction and percutaneous fixation with Kirschner wires (74 children). We also used closed reduction and application of an above elbow cast (12 children), skeletal traction with fixation after swelling resignation (5 children), and open reduction and internal fixation (4 children). In 90 cases we did not observe any neurovascular disorders, and early functional results were good or satisfactory. 2 cases were associated with transient, postreduction radial nerve palsy. In 2 children surgical intervention was necessary due to external compression of the brachial artery in one case, and contusion with brachial artery spasm in another. In one girl we observed development of Volkmann's contracture (treated surgically with good functional result). The authors conclude that the closed reduction and percutaneous K-wire fixation can be used as a treatment of choice for displaced supracondylar fractures of the humerus in children. PMID- 12884654 TI - [Operative treatment of clavicle fractures in children]. AB - We studied retrospectively, 23 fractures of the clavicle in children treated by K wire stabilization between 1999 and 2001. We performed open reduction in 22 children. In one child percutaneous pinning was possible. The anatomical reduction was achieved in all cases. The postoperative course was uneventful in all cases, we didn't notice any infectious complication. The irritation of skin above the protruding end of pin was the only complaint. The pins were removed after 2-4 months. No any additional immobilisation was applied. PMID- 12884655 TI - [Treatment methods for complications of forearm diaphyses fractures, first stabilized with plates]. AB - A short series 8 patients with complications and failures of forearm shafts fractures treated with screws and plates was analysed. There were 2 pseudoarthroses of ulna, delayed union of radial shafts in 4 cases in which 2 had plates loosening. The additional 2 patients had the refractures of radius shortly after metal removal. The intraoperative investigation presented that main cause of delayed unions and pseudoarthroses was insufficient blood supply to the bony ends due to circulation destruction during plates implantation. All 8 patients had finally obtained union after second stage procedure. It was consisted of plates and screws removal, refreshing of fragments ends, cancerous auto grafts implantation and plaster cast immobilization. Intramedullary stabilization with Rush pins was performed in 6 patients. The additional 2 had such wide bone destruction after primary plates stabilization that intramedullary fixation was risky. PMID- 12884656 TI - [Anterior cruciate ligament reconstruction by autogenous bone-patellar tendon bone graft: correlation between bone-tunnel position and clinical results]. AB - The authors evaluate correlation between bone-tunnel position and clinical results of 50 patients, aged 18-47 years, after anterior cruciate ligament reconstruction by bone-patellar tendon-bone autograft with half-tunnel technique. The evaluation comprises the femoral tunnel placement at the lateral femoral condyle, the angle of the tibial tunnel to the medial joint line of the tibia and paraarticular location of the tibial tunnel width clinical outcome within one to five years prior to operation. Bone-tunnel placement was based on radiograms done in anteroposterior and lateral view. Clinical evaluation was based on the International Knee Documentation Committee (IKDC) scale. Statistical analysis was performed by using Fisher exact test, Mann-Whitney test and chi 2 test with Yates correction. The results show that placement the femoral tunnel anteriorly to the line which was perpendicular to the Blumensaat line where it starts to curve posteriorly and tibial tunnel angle of 75 degrees or more is associated with statistically significant worsened of clinical outcome. Malalignment of femoral tunnel placement characterised much larger effects of qualitative and quantitative changes. PMID- 12884657 TI - [Operative treatment of Haglund's deformity]. AB - The paper presents results of the operative treatment of 22 heel in 18 patients mostly young women from 13 to 31 years old (average 20.6) with Haglund's exostosis. Operative technique consisted in partial resection of the heel with removal of the Achilles bursa. The most valuable was radiological examination for the diagnosis and assessment operative results. The ultrasound examination was helpful of estimation of Achilles tendon and calcaneal bursa. Good results achieved in 16 patients (20 heel) with follow-up from 4 to 20 years (average 9.5). Unsatisfactory results in 2 patients was connected with inadequate bone resection. PMID- 12884658 TI - [Surgical treatment of herniated intervertebral discs in children and teenagers]. AB - The authors present early results od surgical treatment of herniated intervertebral discs in children and teenagers. Five cases of herniated intervertebral discs in the lumbar spine in teenagers were treated surgically between 2000 and 2002. The paper presents early results of surgical treatment, as well as the indications for it. It is the authors' opinion that the indications for surgery of herniated lumbar discs among children and teenagers do not differ form indications among adults. Postponing surgery because of young age seems to be unjustified. PMID- 12884659 TI - [Professor Franciszek Raszeja MD--60th anniversary of his death]. AB - The authors present a concise biography of professor Franciszek Raszeja on the sixty years of his death. Special attention is given to his scientific and organizational achievements, particularly in regard to the formation of the Orthopaedic Department at the University of Poznan. PMID- 12884660 TI - [Arthroscopy of the knee joint in the last twenty five years]. AB - This study presents, based on the literature data and own experience, the development of the knee joint arthroscopy in the years of 1975-2002 and its current trends. PMID- 12884661 TI - [Bipolar radial head prosthesis--own design]. AB - The authors describe its own design--bipolar radial head prosthesis. The value of this prosthesis is analyzed. The early results in 10 patients, age 20-69 (average 36 years), 7 male and 3 female have been reported. The results are excellent and good. The authors conclude that this prosthesis should be used as an alternative for radial head resection. PMID- 12884662 TI - [A new distally pedicled adipocutaneous perforator forearm flap]. AB - A new distally pedicled adipocutaneous perforator forearm flap useful in corrections of congenital anomalies of hand was described. Due to the congenital anomaly expression the perforators may origin from radial, median, ulnar or anterior interosseous artery which supply blood flow into the flap tissues. Raising the flap from anterior and radial surface of the forearm begins from the proximal to the distal part, above the fascia leaving superficial cutaneous nerves and cephalic vein intact. The flap may be safely incised in the ratio 5:1 and the pivot point is localized in the place where perforators pierce the forearm fascia usually 1-2 cm above the rascetta. Rerouting of the flap distally on the hand makes reconstruction of missing thenar, the first web space or even finger tissues possible. These conditions are basal for further reconstruction in advanced hypoplasia of the radial part of hand. Similar flap may be used in the posttraumatic cases when the flap blood supply comes exclusively from radial artery perforators. PMID- 12884663 TI - [Surgical approach through the femoral neck in pathological changes of the femoral head]. AB - Three cases patients with tumors or pathological changes of femoral head. They were treated by surgery by femoral neck approach. We reach the tumor without destroy any articular structures of the hip. In two cases the tumors were removed in total, coagulated and filed up with autogenic bone graft. Third patient with traumatical necrotic changes was treated by autograft. PMID- 12884664 TI - [Spontaneous anterior atlanto-axial subluxation--a case story]. AB - In this study, a case of 4.5 years' old girl with spontaneous dislocation of C1/C2 vertebrae probably due to congenital defect of C1-hypoplasia of the atlas anterior arch is presented. Treatment consisted of skull traction and slow reposition followed by surgical spinodesis C1 and C2. During examination after 1 year follow up period, patient was in good clinical condition, without any complaints. On radiograms the cervical spine was stable and of correct shape. PMID- 12884665 TI - [Reconstructive microsurgery procedures in orthopaedic departments performed by plastic surgeon]. AB - The author carried out 50 free flaps in 49 patients during 10 year consulting practice in orthopaedic departments in the regional hospital in Otwock. 43 flaps were transplanted to lower extremities and 7 to upper. Extensive bone defects (bd), requiring reconstruction of shafts especially in case of tibiae predominated. The recipient sites of free tissue flaps were as follows: the arm- 3 (bd), forearm--2 (bd), hand--2 (thumb defects), thigh--6 (bd--5, aseptic necrosis of the femoral head--1), tibiae--33 (bd and nonunion of tibiae--29, soft tissue defects (std)--4), foot--4 (std). In the post-traumatic cases criteria for operations were determined on the basis of the arteriography in the impaired extremity. The operations were performed in the period from some weeks to several years after the trauma. The following free flaps were used: peroneal flap--28, iliac flap--13, radial forearm flap--2, latissimus dorsi flap--3, rectus abdominis muscle flap--1, groin flap with aponeurosis of the external oblique muscle of the abdomen--1, toe--1, composite great and second toe--1. The operating microscope was usually used during microsurgical anastomoses, and in two cases a magnifying glass was used. Standard Ethilon sutures 10-0 with the needle of 100 or 130 microns in diameter were used for anastomoses. End-to-side microanastomoses both for artery and comitant vein prevailed. Dextran 40,000 and Aspirin were administered postoperatively. There was a survival rate of 98 percent with 49 flaps surviving and 1 failing. Early vascular complications were observed in 3 patients (2 in radial flaps and 1 in latissimus dorsi flap). Revision of the arterial microanastomoses was carried out in 2 patients, of the venous in one, obtaining complete survival of one of the flaps. Partial loss of the flap tissue without essential effect on the final treatment was observed in two remaining flaps. One patient with the disturbances of outflow was successfully treated with Heparin administered subcutaneously to the island of the flap. Thrombosis resulting in complete loss of one of the fibular flaps due to the wound infection was observed on the seventh day after the operation. Soft tissue and periosteum around sequestrum were revascularized generating sufficient bone recovery. Different depth necrosis of the island tissue in its peripheral parts was observed in some patients because of difficulties of blood outflow from the flap. Necrectomy or excision of the whole skin island in cases of very fatty flaps and skin grafting were indicated as giving a better cosmesis. Secondary procedures modeling the flap were not performed. Complete rebuilding of tibiae was not obtained only in two children treated because of congenital pseudoarthrosis of tibiae. In one patient with femur defect and vascularized fibular bone reconstruction late result of treatment is unknown. Treatment of the remaining patients was successful (92 percent). PMID- 12884666 TI - In search of global leaders. AB - For all the talk about global organizations and executives, there's no definitive answer to the question of what we really mean by "global." A presence in multiple countries? Cultural adaptability? A multilingual top team? We asked four CEOs and the head of an international recruiting agency--HSBC's Stephen Green, Schering Plough's Fred Hassan, GE's Jeffrey-lmmelt, Flextronics's Michael Marks, and Egon Zehnder's Daniel Meiland--to tell us what they think. They share some common ground. They all agree, for example, that the shift from a local to a global marketplace is irreversible and gaining momentum. "We're losing sight of the reality of globalization. But we should pay attention, because national barriers are quickly coming down", Daniel Meiland says. "If you look ahead five or ten years, the people with the top jobs in large corporations ... will be those who have lived in several cultures and who can converse in at least two languages." But the CEOs also disagree on many issues--on the importance of overseas assignments, for instance, and on the degree to which you need to adhere to local cultural norms. Some believe strongly that the global leader should, as a prerequisite to the job, live and work in other countries. As Stephen Green put it, "If you look at the executives currently running [HSBC's] largest businesses, all of them have worked in more than one, and nearly all in more than two, major country markets." Others downplay the importance of overseas assignments. "Putting people in foreign settings doesn't automatically imbue new attitudes, and it is attitudes rather than experiences that make a culture global," says Fred Hassan. The executives' essays capture views that are as diverse and multidimensional as the companies they lead. PMID- 12884667 TI - Microcapitalism and the mega-corporation. AB - More than 100 miles from Bangalore, India, there's a rural area called Kuppam where one in three citizens is illiterate, more than half of the households have no electricity, and there's a high rate of AIDS. It's exactly this challenging atmosphere that prompted Hewlett-Packard to choose Kuppam as one of its first "i communities" initiatives. Through the program, HP creates public-private partnerships to accelerate economic development through the application of technology while simultaneously opening new markets and developing new products and services. HP brings to these initiatives the management disciplines of a successful technology business. For example, it unearths customer needs using an iterative cycle that involves prototyping products and services and then closely observing residents' experiences with them. It fields a diversely talented team that brings many skills to the initiative, including deep technical ability, management acumen, and market knowledge. It takes a systems approach, simultaneously examining all the elements that must come together to create a working solution to a given problem. It establishes a "leading platform" on which other players--comapnies, nonprofits, and government agencies--can build technologies and applications. Practices like these help ensure that HP's investment yields real, sustainable results for the community in question. But HP also sees returns to its own business. In Kuppam, the company is discovering the need for (and developing) new products like a solar-powered digital camera, with printer, that fits in a backpack. By engaging the community and its leaders and working with them to design valuable new tools and capabilities, HP is gaining the knowledge it needs to be a stronger competitor in other developing regions. PMID- 12884668 TI - Abraham Lincoln and the global economy. AB - Abraham Lincoln would have well understood the challenges facing many modern emerging nations. In Lincoln's America, as in many developing nations today, sweeping economic change threatened older industries, traditional ways of living, and social and national cohesion by exposing economies and societies to new and powerful competitive forces. Yet even in the midst of the brutal and expensive American Civil war--and in part because of it--Lincoln and the Republican Congress enacted bold legislation that helped create a huge national market, a strong and unified economy governed by national institutions, and a rising middle class of businessmen and property owners. Figuring out how to maximize the benefits of globalization while minimizing its disruptions is a formidable challenge for policy makers. How do you expand opportunities for the talented and the lucky while making sure the rest of society doesn't fall behind? It may be helpful to look at the principles that informed the policies that Lincoln and the Republican Congress instituted after they came to power in 1861: Facilitate the upward mobility of low- and middle-income groups to give them a significant stake in the country. Emphasize the good of the national economy over regional interests. Affirm the need for sound government institutions to temper the dynamics of the free enterprise system. Tailor policies to the national situation. Realize that a period of turmoil may present a unique opportunity for reform. These principles drove the reforms that helped Americans cope with and benefit from rapid technological advances and the fast integration of the American economy in the nineteenth century. They may be instructive to today's policy makers who are struggling to help their own citizens integrate into the fast-changing global economy of the twenty-first century. PMID- 12884669 TI - The new world disorder. AB - On January 1, 1995, representatives from 76 countries signed the World Trade Organization charter, which for years had been part of a temporary trade agreement. The WTO's emergence as a fully empowered supranational body seemed to reflect the triumph of what the first President Bush had described as the "new world order." That order was based on two assumptions: that a healthy economy and a sound financial system make for political stability, and that countries in business together do not fight each other. The number one priority of U.S. foreign policy was thus to encourage the former Communist countries of Europe and the developing nations in Latin America, Asia, and Africa to adopt business friendly policies. Private capital would flow from the developed world into these countries, creating economic growth. It sounded too good to be true, and so it proved. The new world order of Bush pere and his successor, Bill Clinton, has been replaced by the new world disorder of Bush fils. Under the second Bush's administration, the economic and political rationale-behind the Washington consensus of the 1990s has unraveled, forcing a radical change in our perceptions of which countries are safe for business. Negotiating this new environment will require companies to more rigorously evaluate political events and more carefully assess the links between political, economic, and financial risk factors. They'll need to be more selective about which markets to enter, and they'll need to think differently about how to position themselves in those markets. The geopolitical events of the past year, the Bush administration's global war on terror, as well as ongoing convulsions in traditional political and economic relationships must be understood and managed by corporate leaders worldwide. With careful analysis, business leaders can increase their companies' visibility and better respond to the uncertainties of the new world disorder. PMID- 12884670 TI - What is a global manager? AB - Riven by ideology, religion, and mistrust, the world seems more fragmented than at any time since, arguably, World War II. But however deep the political divisions, business operations continue to span the globe, and executives still have to figure out how to run them efficiently and well. In "What Is a Global Manager?" (first published in September-October 1992), business professors Christopher Bartlett and Sumantra Ghoshal lay out a model for a management structure that balances the local, regional, and global demands placed on companies operating across the world's many borders. In the volatile world of transnational corporations, there is no such thing as a "universal" global manager, the authors say. Rather, there are three groups of specialists: business managers, country managers, and functional managers. And there are the top executives at corporate headquarters who manage the complex interactions between the three--and can identify and develop the talented executives a successful transnational requires. This kind of organizational structure characterizes a transnational rather than an old-line multinational, international, or global company. Transnationals integrate assets, resources, and diverse people in operating units around the world. Through a flexible management process, in which business, country, and functional managers form a triad of different perspectives that balance one another, transnational companies can build three strategic capabilities: global-scale efficiency and competitiveness; national-level responsiveness and flexibility; and cross-market capacity to leverage learning on a worldwide basis. Through a close look at the successful careers of Leif Johansson of Electrolux, Howard Gottlieb of NEC, and Wahib Zaki of Procter & Gamble, the authors illustrate the skills that each managerial specialist requires. PMID- 12884671 TI - The end of corporate imperialism. AB - As they search for growth, multinational corporations will have no choice but to compete in the big emerging markets of China, India, Indonesia, and Brazil. But while it is still common to question how such corporations will change life in those markets, Western executives would be smart to turn the question around and ask how multinationals themselves will be transformed by these markets. To be successful, MNCs will have to rethink every element of their business models, the authors assert in this seminal HBR article from 1998. During the first wave of market entry in the 1980s, multinationals operated with what might be termed an imperialist mind-set, assuming that the emerging markets would merely be new markets for their old products. But this mind-set limited their success: What is truly big and emerging in countries like China and India is a new consumer base comprising hundreds of millions of people. To tap into this huge opportunity, MNCs need to ask themselves five basic questions: Who is in the emerging middle class in these countries? How do the distribution networks operate? What mix of local and global leadership do you need to foster business opportunities? Should you adopt a consistent strategy for all of your business units within one country? Should you take on local partners? The transformation that multinational corporations must undergo is not cosmetic--simply developing greater sensitivity to local cultures will not do the trick, the authors say. To compete in the big emerging markets, multinationals must reconfigure their resources, rethink their cost structures, redesign their product development processes, and challenge their assumptions about who their top-level managers should be. PMID- 12884672 TI - Thriving locally in the global economy. AB - More and more small and midsize companies are joining corporate giants in striving to exploit international growth markets. At the same time, civic leaders worry about their communities' economic future in light of the impact of global forces on the operation and survival of business. How can communities retain local vitality yet still link their business to the global economy? Harvard professor Rosabeth Moss Kanter addresses that question in this classic HBR article, orginally published in 1995. To avoid a clash between international economic interests and local political interests, globalizing business must learn how to be responsive to the communities in which they operate, Kanter says. And communities must determine how to create a civic culture that will attract and retain footloose companies. The author surveyed five U.S. regions with direct connections to the global economy--Boston, Cleveland, Miami, Seattle, and the Spartanburg-Greenville region of South Carolina--to determine their business and civic leader's strategies for improving their constituent's quality of life. She identified ways in which the global economy can work locally by capitalizing on the resources that distinguish one place from another. Kanter argues that regions can invest in capabilities that connect their local populations to the global economy in one of three ways: as thinkers, makers, or traders. She points to the Spartanburg-Greenville region as a good example of a world-class makers, with its exceptional blue-collar workforce that has attracted more than 200 companies from 18 countries. The history of the economic development of this region is a lesson for those seeking to understand how to achieve world-class status and bring local residents into the world economy. PMID- 12884673 TI - Turn public problems to private account. AB - Many managers face increasing calls to invest corporate resources in charitable causes. How should executives balance a firm's very real economic imperative to maximize profitability with its hypothetical moral imperative to improve society? To provide one answer, the author draws on his experience as president of an economic-development company, IBEC. Viewing profit as "an essential discipline and measure of economic success" but not "the sole corporate goal," the company actively invested in social programs that met four criteria: they served a need of the local population; they required innovative approaches; they made sense on economic grounds; and they respected the social norms of the community. Such civic-minded efforts, the author argues in this prescient 1971 article, not only improve people's lives but also create the foundation for more affluent and dynamic markets--markets that ultimately produce greater profits for business. For example, one of IBEC's earliest ventures was directed toward solving Venezuela's problems in retail food marketing. Many important items were unavailable at the small stores where people shopped. So in 1949, working with local partners, IBEC opened a supermarket. Supermarkets soon changed the food buying habits of the nation, and the initiative helped alter patterns of food distribution and created the reliable demand needed to establish a host of local suppliers. Return on IBEC's investment, and that of its local partners, was most satisfactory, the author reports. The road to meeting a public need-especially a major one--is rarely easy, the author says. But if management sizes up the need well, there is a good chance its new venture will survive under adversity. PMID- 12884674 TI - Food for thought. PMID- 12884675 TI - Women vs. Wal-Mart. PMID- 12884676 TI - Bioplastic fantastic. PMID- 12884677 TI - Backward conditioning: mediation by the context. AB - The information acquired in backward conditioning (i.e., outcome-->cue) was assessed in 3 Pavlovian lick-suppression experiments with water-deprived rats as subjects. Experiment 1 confirmed previous research that few outcome-->cue pairings made the cue into a conditioned excitor and additionally showed that massive posttraining extinction of the training context attenuated a backward trained cue's excitatory value. Experiment 2 found that many outcome-->cue pairings made the cue into a conditioned inhibitor and that the same context manipulation attenuated this inhibitory value. Experiment 3 confirmed the observations of Experiments 1 and 2 and demonstrated that these effects of context extinction were specific to backward-trained cues conditioned in the extinguished context. These results are interpreted in terms of cue-->context and context-->outcome associations. PMID- 12884678 TI - Abstract-concept learning and list-memory processing by capuchin and rhesus monkeys. AB - Three capuchin monkeys (Cebus apella) touched the lower of 2 pictures (same) or a white rectangle (different), increased same/different abstract-concept learning (52% to 87%) with set-size increases (8 to 128 pictures), and were better than 3 rhesus monkeys (Macaca mulatta). Three other rhesus that touched the top picture before choices learned similar to capuchins but were better at list-memory learning. Both species' serial position functions were similar in shape and changes with retention delays. Other species showed qualitatively similar shape changes but quantitatively different time-course changes. In abstract-concept learning, qualitative similarity was shown by complete concept learning, whereas a quantitative difference would have been a set-size slope difference. Qualitative similarity is discussed in relation to general-process versus modular cognitive accounts. PMID- 12884679 TI - Modularity as a fish (Xenotoca eiseni) views it: conjoining geometric and nongeometric information for spatial reorientation. AB - When disoriented in a closed rectangular tank, fish (Xenotoca eiseni) reoriented in accord with the large-scale shape of the environment, but they were also able to conjoin geometric information with nongeometric properties such as the color of a wall or the features provided by panels located at the corners of the tank. Fish encoded geometric information even when featural information sufficed to solve the spatial task. When tested after transformations that altered the original arrangement of the panels, fish were more affected by those transformations that modified the geometric relationship between the target and the shape of the environment. Finally, fish appeared unable to use nongeometric information provided by distant panels. These findings show that a reorientation mechanism based on geometry is widespread among vertebrates, though the joint use of geometric and nongeometric cues by fish suggest that the degree of information encapsulation of the mechanism varies considerably between species. PMID- 12884680 TI - Linking of serially ordered lists by macaque monkeys (Macaca mulatta): list position influences. AB - Five monkeys (Macaca mulatta) were trained on 2 sets of 3 5-item serially ordered lists. Then, each set was either linked or not in a counterbalanced, within subject design. Linking entailed training on the 2 pairs that ordered the 3 5 item lists into a single overall 15-item series. Choices on novel pairings after linking conditions attempted to define the unique contributions of knowledge of within-list ordinal position and between-lists link training. With linkage, the series was immediately treated as a 15-item ordered list. Without linkage, choices reflected list positions from initial learning, but continued testing with directional reward yielded gradual ordering into a 15-item list. Apparently, monkeys remembered and used initial list-position information, but linkage allowed inference of an integrated serial relationship among items. Results supported primate list memory as an organizational process. PMID- 12884681 TI - Anterograde amnesia for Pavlovian fear conditioning and the role of one-trial overshadowing: effects of preconditioning exposures to morphine in the rat. AB - Four experiments studied anterograde deficits in Pavlovian fear conditioning following prolonged exposure to the mu-opioid receptor agonist morphine. Injections of morphine produced temporally graded anterograde amnesia characterized by deficits in contextual and conditioned-stimulus (CS) conditioning 1 or 7 days and selective impairment in CS conditioning 21 days after last injection. This anterograde deficit in conditioning did not recover across a retention interval, was absent when rats were tested immediately after conditioning, and required the presence of an auditory CS. These results suggest that anterograde deficits in Pavlovian fear conditioning emerged from differences in susceptibility to 1-trial overshadowing of context by CS. PMID- 12884682 TI - How do tufted capuchin monkeys (Cebus apella) understand causality involved in tool use? AB - Four tufted capuchin monkeys (Cebus apella) were trained to choose from 2 hook like tools, 1 of which successfully led to collecting food, whereas the other did not because of inappropriate spatial arrangement of the tool and the food. In Experiment 1, all of the monkeys successfully learned the basic task. The monkeys performed successfully with tools of novel colors and shapes in Experiments 2-5. These results demonstrate that the monkeys used the spatial arrangement of the tool and the food as a cue. However, they failed when there were obstacles (Experiment 6) or traps (Experiment 7) on the path along which the monkeys dragged tools. These results may suggest that capuchin monkeys understand the spatial relationship between 2 items, namely, food and the tool, but do not understand the spatial relationship among 3 items, namely, food, tool, and the environmental condition. The possible role of stimulus generalization is also considered. PMID- 12884683 TI - Comparing the properties of neuronal culture systems: a shopping guide for the cell biologist. AB - Cell biologists of many stripes may find that their question of interest can be studied to advantage in neurons. However, they will also find that "neurons" include many and diverse cell types among which perhaps just one or a few may be ideal for a particular experiment. This chapter discusses the properties, relative complexity, and cost of primary neurons and neuronal cell types from different species and parts of the nervous system and compares their utility for different kinds of cell biological experiments. PMID- 12884684 TI - Growing and working with peripheral neurons. AB - Cultures of vertebrate peripheral neurons have been used to address a variety of issues related to the cell biology of the neuron. They are particularly amenable to experimental manipulations, such as microinjection, and can be cultured under a variety of different conditions designed to meet the needs of the particular experiment. This chapter focuses on cultures of rat sympathetic neurons from the superior cervical ganglia and on cultures of chick sensory neurons from the lumbosacral dorsal root ganglia. Information is provided on methods for dissection, preparation of culture dishes and substrates, composition of media, relevant growth factors, reduction of nonneuronal contamination, and maintenance of the cultures. PMID- 12884685 TI - Dissection and culturing of chick ciliary ganglion neurons: a system well suited to synaptic study. AB - This chapter describes the function and development of the ciliary ganglion, the potential of ciliary ganglion neurons as a cell biological tool, and their dissection, dissociation, and culturing. Ciliary ganglion neurons grow unusually rapidly on a laminin-based substratum and develop large, thin calyx terminals in culture in less than 12 h. The two neuronal classes present in the cultures can be identified by size alone. The limited number of ganglia per animal renders this ganglion a poor choice for biochemical studies based on the extraction of cultured cells. However, they are ideally suited for studies based on single-cell observation, particularly investigation of presynaptic mechanisms using fluorescence microscopy. PMID- 12884686 TI - The culture of chick forebrain neurons. AB - Dissociation of the forebrain of a single 8-day chick embryo produces > 10(7) neurons in nearly pure culture. Our methods allow 50-70% of these neurons to develop an axon and typical pyrimidal shape after 3-4 days in culture at low density (10(4) cells/cm2) by a stereotyped developmental sequence similar to that of rat hippocampal neurons. The culture method for chick forebrain neurons is unusually rapid, inexpensive, simple, and could be used in undergraduate laboratory exercises. The dissection and dissociation of the tissue are easy and rapid, requiring less than 30 min from cracking open the chicken egg to plating the cells. Axonal development by these neurons and growth for about a week do not require glial support. The neurons are grown on polylysine-treated culture surfaces in either CO2-dependent (Medium 199) or -independent (Liebovitz L15) media with 10% fetal bovine serum and a supplement based on the classic N2 supplement for neuronal culture. PMID- 12884687 TI - Growing and working with spinal motor neurons. AB - The chick embryo has a long tradition as a model organism in developmental biology as well as embryology. A year-round supply of fertilized eggs, accessibility to all stages of development, and the ease of manipulation of the embryo all contribute to the advantages of investigations using chick embryos. A plethora of culture systems have been developed over the past century allowing to culture intact embryos from as early as 2 days of development. Other culture systems include whole embryo slices, organotypic cultures, tissue explants, and dissociated cultures. Studies utilizing the chick embryo, and in particular spinal motor neurons, were crucial for our present knowledge of the development but also adult physiology, injury, and disease of the nervous system. Extensive studies on spinal motor neurons revealed many molecular mechanisms underlying fundamental events, such as neural induction, axon guidance, programmed cell death, and neuron-target interaction. Cultures of dissociated spinal motor neurons represent one important experimental paradigm. This chapter describes two alternative procedures to establish dissociated spinal motor neuron cultures with virtually no contamination by nonneuronal cells. PMID- 12884688 TI - Avian Purkinje neuronal cultures: extrinsic control of morphology by cell type and glutamate. AB - An in vitro coculture system is described to study the avian Purkinje neuron and the interactions occurring with astrocytes and granule cells during development in the cerebellum. Astrocytes initially and granule cells later regulate Purkinje neuron morphology. The coculture system presented here provides an excellent system for investigating the morphological, immunocytochemical, and electrophysiological differentiation of Purkinje neurons under controlled conditions and for studying cell-cell interactions and extrinsic factors, e.g., glutamate in normal and neuropathological conditions. PMID- 12884689 TI - Culturing hippocampal and cortical neurons. AB - Primary cultures of rat and mouse hippocampus and cerebral cortex are widely used to study neuronal properties such as axonal extension, synaptic transmission, and excitotoxicity. Short-term culturing of these neurons can be very straightforward and is perhaps easier than culturing cells lines once dissections are made and cell stocks are frozen. Long-term cultures of relatively pure neuronal populations require slightly more effort, but protocols are described that are less complicated than most published protocols. These include simpler ways to clean and coat coverslips, as well as using glia-conditioned medium to eliminate the need to make individual cocultures of neurons and glia. These methods consistently yield hippocampal and cortical cultures expressing dendritic spines and synapses that survive over 3 weeks in culture. For investigators employing biochemical assays where a fairly large amount of protein is necessary, cortical neurons may be especially attractive to use as large amounts of tissue are obtained and available for culture. PMID- 12884690 TI - Working with Xenopus spinal neurons in live cell culture. AB - Neurons from the Xenopus spinal cord are highly versatile and easily manipulated, making them an ideal model system to answer questions regarding the cellular and molecular basis of early neural development and function. Xenopus has been a productive model system in studies ranging from axon growth and guidance to synaptic plasticity. Exogenous molecules, such as proteins, fluorescent tracers, and nucleic acids, can be injected into early blastomeres to load tracers in all neurons or into late blastomeres to target specific classes of neurons based on established lineage maps. Xenopus spinal neurons also provide an excellent culture system, as neurons extend processes on a variety of substrata and develop at room temperature in minimal salt solutions. Live fluorescent neurons can be imaged for hours with fluorescence microscopy at room temperature in static cultures without neurotrophic support or serum. This highly reduced culture system minimizes variables that can confound interpretation of results. Cultures can be prepared at various stages of development as dissociated neurons or as spinal cord explants. Both excitatory and inhibitory neurons develop in culture, and synaptic contacts among neurons and between neurons and nonneuronal targets form naturally. The simple anatomy and rapid rostral-to-caudal development of the Xenopus spinal cord also make this an excellent in vivo model system to analyze axon guidance by identifiable classes of neurons. This chapter focuses on techniques that exploit both in vitro and in vivo qualities of this system. PMID- 12884691 TI - Culturing neurons from the snail Helisoma. AB - The large neurons of the freshwater snail Helisoma trivolvis provide a unique preparation to study cytoskeletal mechanisms involved in neuronal growth and axon guidance. When placed into culture, these neurons form large growth cones in which cytoskeletal components and their dynamics can be analyzed with high spatial resolution. Moreover, these growth cones display all of the dynamic features characteristic of growing axons, including advance, pause, collapse, and turning, allowing the correlation of cell biological mechanisms with growth cone motility. This chapter describes complete procedures for culturing Helisoma neurons, including snail dissection, enzymatic treatments, removal of neurons, and necessary solutions, equipment, and supplies. Techniques are presented to culture Helisoma neurons by the extraction and transfer of individual neurons to culture dishes. A newer technique to dissociate neurons from whole ganglia is also described. In addition, methods to culture neurons on two substrates are presented. Culturing on polylysine in defined medium produces large, but nonmotile growth cones for cytoskeletal analysis, whereas culturing on polylysine in conditioned medium allows growth and motility for behavioral analysis. Recent tests suggest a new, simpler formulation for the medium used to culture Helisoma neurons that does not require the special-order medium that was previously used for cultures. These procedures make it feasible for someone inexperienced to successfully culture Helisoma neurons for use in a variety of experiments. PMID- 12884692 TI - The tibial-1 pioneer pathway: an in vivo model for neuronal outgrowth and guidance. AB - As neurons extend axons to their targets during development, growth cones must reorient their direction of migration in response to extracellular guidance cues. A variety of model systems have been employed in order to dissect the cellular and molecular mechanisms that underlie this complex process. One preparation, the developing grasshopper limb bud, has proved to offer a number of advantages in which to examine mechanisms of growth cone guidance and motility in vivo. First, the relatively large size of the embryonic nervous system allows for straightforward imaging of both fixed and live neurons in vivo. Second, the peripheral nerves generated in the limb bud are highly stereotyped. Third, intact embryos can be cultured for a period of days, allowing for fairly easy perturbations at precise developmental stages. Fourth, due to the ease of dissection, numerous cell biological and molecular techniques can be utilized in the limb bud. Finally, axon guidance molecules and mechanisms are conserved between grasshoppers and other organism, including vertebrates. PMID- 12884693 TI - Techniques to dissect cellular and subcellular function in the Drosophila nervous system. PMID- 12884694 TI - PC12 cells as a model for studies of regulated secretion in neuronal and endocrine cells. AB - Pheochromocytoma-derived cell lines such as PC12 cells maintain a differentiated neuroendocrine phenotype and have been widely used as a convenient model system for a wide variety of cell biological studies on neurotrophin action, monoamine biogenesis, protein trafficking, and secretory vesicle dynamics. This chapter reviews a number of methods that are useful for studies of the regulated dense core vesicle secretory pathway. This includes protocols for maintaining cells and preserving their phenotype. A variety of assays are discussed for monitoring secretion in intact or permeable cells and in transfected cells. Specific methods for immunocytochemical studies in permeable cells are discussed. Finally, protocols for high-efficiency PC12 cell transfections and the isolation of stably transfected cell lines are provided. PMID- 12884695 TI - B35 neuroblastoma cells: an easily transfected, cultured cell model of central nervous system neurons. AB - A panel of neuronal cell lines was derived from tumors of the neonatal rat central nervous system (CNS) in 1974, and two of these lines are in wide use today. Both the B35 and B50 lines offer a number of advantages to researchers who study CNS neurons in culture: they are simple to grow, to differentiate, and to transfect. B50 cells have been used extensively in the study of neuronal cell death, toxicology, and differentiation, whereas B35 cells have proven useful in the molecular analysis of endocytosis and of signaling pathways, in particular those that guide axonal outgrowth and cell motility. This chapter provides protocols for growing and transfecting B35 cells, selecting stable transfectants, exploring protein function using an antisense approach, and assaying cell motility in a Transwell chamber. All of these protocols have been written for researchers who have some skill in basic cell culture techniques, but previous experience with cultured neurons is not required. PMID- 12884697 TI - Making proteins into drugs: assisted delivery of proteins and peptides into living neurons. AB - The introduction of exogenous proteins and peptides into cells is a valuable experimental approach. However, exogenous proteins are not internalized by living cells readily. This limitation has been overcome by the development of peptide based methods that assist in the delivery of exogenous proteins into the cytoplasm and nucleus. These methods may facilitate the in vivo delivery of reagents in therapies aimed at neurodegenerative disorders and recovery from nervous system injury. PMID- 12884696 TI - Live-cell imaging of slow axonal transport in cultured neurons. AB - Cytoskeletal polymers and other cytosolic protein complexes are transported along axons in the slow components of axonal transport. Studies on the movement of neurofilaments and microtubules in the axons of cultured neurons indicate that these polymers actually move at fast rates and that the movements are also infrequent and highly asynchronous. These observations indicate that the slow rate of slow axonal transport is due to rapid movements interrupted by prolonged pauses which presents special challenges for studies on the mechanism of movement. This chapter describes the procedures that the author's laboratory has used to observe and analyze the movement of neurofilaments and microtubules in axons of cultured neurons from the superior cervical ganglia of neonatal rats. In particular, the author describes how to culture these neurons, how to transfect them by nuclear injection, and how to detect the rapid and infrequent movement of cytoskeletal polymers using time-lapse fluorescence imaging. PMID- 12884698 TI - Transfection of primary central and peripheral nervous system neurons by electroporation. AB - Neurons are difficult cells to transfect. Many methods that work routinely for immortalized tissue culture cells or primary cultures of nonneuronal cells are ineffective, toxic, or both when applied to neurons. This chapter describes a protocol that optimizes electroporation-based transfection of chick embryonic peripheral and central neurons. The key features required for successful electroporation and recovery of transfected neurons are high cell density, correct applied voltage and pulse duration, and the presence of calcium ions in the electroporation medium. Less important features are temperature, postporation rest, and the general composition of the electroporation medium. We emphasize the rationale for each element in our method and provide information useful for optimizing the procedure for other neurons. PMID- 12884699 TI - Biolistic transfection. AB - Recent improvements in the biolistic technique and devices have increased its usefulness for transfection of neurons. With these recent advances, both dissociated and slice cultures can be transfected at reasonably high rates. This chapter focuses on the parameters that determine the successful biolistic transfection of neurons in both types of cultures. PMID- 12884700 TI - Expression of transgenes in primary neurons from chick peripheral and central nervous systems by retroviral infection of early embryos. AB - Many cell biological studies require the expression of transgenes in cells in culture, but it is difficult to obtain uniform, stable, and efficient expression of transgenes in primary neurons. We have approached this problem by adapting from developmental biologists the avian retroviral vector, RCAS. This vector allows the introduction of a transgene by infection early in chick embryonic development. Transgenes that are less than 2.6 kb in size can be cloned through an adapter vector, SLAX 12 NCO, and into the RCAS retroviral vector with relative ease. The vector is then used to produce active virus, and the virus is injected into the neural tube or ventricles of stage 10 embryos. By infecting the neuronal precursor cells while they are still mitotic, the retrovirus and accompanying transgene are introduced into the genome and subsequently spread by replication, shedding of new virus, and infection of other cells. Embryos are incubated from the time of injection until E9-E12 and peripheral and central nervous system neurons are dissected out and grown in culture using standard techniques. In this manner, the majority of the sympathetic and dorsal root ganglion neurons can be induced to express the trangene. A similar result, at lower efficiencies, is obtained for central nervous system neurons. PMID- 12884701 TI - Production and use of replication-deficient adenovirus for transgene expression in neurons. AB - Adenoviruses infect a wide range of cell types, do not require integration into the host cell genome, and can be produced as replication-deficient viruses capable of expressing transgenes behind any desired promoter. Thus, they are ideal for use in expressing transgenes in the postmitotic neuron. This chapter describes simplifications in the protocols for making recombinant adenoviruses and their use in expressing transgenes in primary neurons of several different types. PMID- 12884702 TI - It's more than just Tenet. Analysis shows not-for-profit hospitals, including a cluster in New Jersey, also heavily rely on outlier payments. AB - After using a loophole in the Medicare outlier payment program to fuel growth, Tenet Healthcare Corp. has found itself operating under a financial cloud. But the spotlight hasn't focused on many other hospitals and systems that have relied heavily on outlier payments to boost their bottom lines. At left, Saint Barnabas Medical Center in Livingston, N.J., ranks among hospitals with a high reliance on outlier payments. PMID- 12884703 TI - Experimenting with quality. CMS-Premier initiative to reward best, punish worst. PMID- 12884704 TI - What suppliers think. GPO supply contracts don't ensure success: survey. PMID- 12884705 TI - Starting over. Docs vow to keep working for malpractice reform. PMID- 12884706 TI - Battle over Medicare. Providers ready to defend their pet provisions. PMID- 12884707 TI - Another membership study. After rejecting sweeping change, AMA to pay for another organizational review. PMID- 12884708 TI - Hard times. Legal fallout for HealthSouth driven by post-Enron climate, observers say. PMID- 12884709 TI - Cloudy forecast. Analysts cite waning admissions as potential threat to not-for profits' credit. PMID- 12884710 TI - Coming together. United Memorial merging with Spectrum in Mich. PMID- 12884711 TI - Too friendly? Teaching hospitals revisit ties to pharmaceutical firms. PMID- 12884712 TI - They've got the blues. N.C. Blues abandons for-profit conversion plan; fines likely at CareFirst. PMID- 12884713 TI - Proof of privacy. Certification offered for HIPAA 'business associates'. PMID- 12884714 TI - The healthcare doctrine. U.S. foreign policy must include the eradication of dangerous diseases. PMID- 12884715 TI - On a mission. Foundation, network work to improve healthcare in Latin America more through building relationships than building facilities. PMID- 12884716 TI - Bloodless coup. Funded by the Army, Oregon researchers turn to the sea to develop a revolutionary bandage that stanches heavy bleeding. PMID- 12884717 TI - How to kill a mandate. Hospital lobby persuades Scully, CMS to scuttle proposal that would have required hospitals to measure patient satisfaction. AB - The Centers for Medicare and Medicaid Services has backed off its plan to require all Medicare-certified hospitals to publicly report mandatory patient surveys, Modern Healthcare has learned. Instead, no regulations will be written and hospitals will be asked to voluntarily use surveys. American Hospital Association President Richard Davidson, left, co-authored a letter urging hospitals to participate. PMID- 12884718 TI - Broadlane's trailblazing mission. San Francisco company's deal reduces tenet stake to that of minority holder. PMID- 12884719 TI - Letting it all hang out. Not-for-profits slow to post finances online. PMID- 12884720 TI - Getting to the bottom. HealthSouth due to offer earnings projections. PMID- 12884721 TI - Resident restrictions. Most teaching hospitals ready for work-hour rules. PMID- 12884722 TI - Overtime threat. Proposed regulation could limit nurses' overtime. PMID- 12884723 TI - Mixed reaction. Molina's IPO goes well, but not all observers cheer. PMID- 12884724 TI - Easing restrictions. Calif. bill would ban service restrictions for NFPs. PMID- 12884725 TI - Bringing in the big guns. U.S. attorney joins civil suits against Medco Health. PMID- 12884726 TI - Mission: control. System, county fight for rights to Tennessee hospital. PMID- 12884727 TI - Triage for overcrowding. Hospitals should fix the emergency department problems they can control. PMID- 12884728 TI - More bills coming due. Although Medicare reform has passed Congress, the final package remains uncertain. And what about the fate of other health legislation? PMID- 12884729 TI - Forging the path. Former Senate aide, lobbyist helps White House craft healthcare policy, reform strategies. PMID- 12884730 TI - In death's shadow. PMID- 12884731 TI - A law's fetal flaw. PMID- 12884732 TI - Burned by SPF. PMID- 12884733 TI - [Function of cortical basal nuclei: Pathophysiology of Parkinson's disease]. PMID- 12884734 TI - [Function of TRP channel as a thermal receptor]. PMID- 12884735 TI - [Detection of anti-early endosome antigen 1 antibody in sera showing cytoplasmic vesicular staining, taken from patients with connective tissue diseases]. AB - An early endosome antigen previously reported by F.T. Mu could be stained on cytoplasmic vesicles of HEp-2 cells. Here, we have investigated autoantibodies against cytoplasmic vesicular antigens, especially against early endosome antigen 1. Twelve sera were selected on the basis of cytoplasmic vesicular staining patterns of HEp-2 cells. Protein-immunoprecipitation using 35S-methionine labeled HeLa lysates, and RNA immunoprecipitation using 32P-labeled HeLa lysates were conducted to characterize the cognate antigens. Nine of 12 sera reacted with proteins in the range of 162-180 kDa, three of which were found to react specifically with the 162 kDa 35S methionine labeled recombinant early endosome antigen 1. These proteins were not associated with common RNA. Although complete clinical information was not available, some of the patients had rheumatoid arthritis(RA). In addition, the RNA-IPP results suggest that other patients included one each with SLE, SSc, polymyositis, and Sjogren's syndrome. Anti-early endosome antigen 1 antibody was found in 25%(3 of 12) of sera known to stain cytoplasmic vesicles. The reactive sera came mostly from patients with RA. The sera was from one case each of clinical-confirmed RA, SLE and Sjogren's syndrome. PMID- 12884736 TI - [A variant hemoglobin found by dissociation of blood glucose from HbA1c on routine physical examination]. AB - Discrepancy between HbA1c measured by routine HPLC and blood glucose was observed in four out of 340 persons on physical examination. We investigated whether this discrepancy was due to abnormal hemoglobin. HbA1c was measured by routine HPLC and latex agglutination. For further examinations, a hemoglobin specimen was analyzed by weak cation-exchange column(PolyCAT A) chromatography, ESI/MS and MS/MS. HbA1c levels measured by HPLC were about half of those measured by latex agglutination. An abnormal peak between HbA1c and HbA0 was detected on HPLC chromatography in three consanguineous persons. The abnormal hemoglobin from one of the related person was also confirmed by PolyCAT A chromatography. The hemoglobin specimen contained almost same amounts of normal HbA0 and abnormal Hb. Analyses of the globin by ESI/MS and MS/MS revealed that glutamate at the 22nd amino acid residue of the beta-chain was replaced by alanine. This variant was the same as HbG-Coushatta[beta 22(B4) Glu-->Ala]. Family studies showed that the variant was inherited as a dominant trait. The dissociation observed in the case was due to a presence of a variant hemoglobin. In the other case HbA1c levels measured by the two methods were similarly low, and no abnormal peak was observed on HPLC chromatography. Similar studies on the hemoglobin of this case did not disclose any abnormalities. The low values of HbA1c of this case were considered in the lowest region of the normal range. PMID- 12884737 TI - [Measurement of plasma von Willebrand factor cleaving protease in patients with varied thrombotic microangiopathy]. AB - Thrombotic thrombocytopenic purpura(TTP) is a multisystem disorders characterized by thrombocytopenia, microangiopathic hemolytic anemia associated with red cell fragmentation, and neurological and renal symptoms. Plasma of patients with TTP has been shown to contain unusually large von Willebrand factor(vWF) multimers that may cause platelet agglutination in vivo. Recently, a metalloprotease responsible for cleavage of vWF multimers has been isolated from normal human plasma and was found to be deficient in some patients with TTP. We examined the activity of the vWF-cleaving protease(vWF-CP), by modified Furlan's method, in plasma from patients with a familial TTP, 3 acquired TTP, 4 thrombotic microangiopathy(TMA) and 2 veno-occlusive disease(VOD) associated after allo-BMT. Diluted plasma samples of patients were incubated with protease-free vWF purified from normal human plasma, in the presence of urea and barium ions. The extent of vWF degradation was assayed by electrophoresis in SDS-agarose gels and immunoblotting. Activity of vWF-CP from 12 normal plasma have been shown as 77 180%(average 115%), whereas, no vWF-CP(below 5%) was observed in plasma from familial TTP, before and after plasma exchange, although FFP infusion therapy has been effective for this patient to recover thrombocytopenia. In 3 acquired TTP, 2 patients showed lack of vWF-CP activity in plasma, and inhibitors against vWF-CP have been elucidated by plasma cross-mixing test. After extensive plasma exchange and FFP infusion followed by corticosteroid therapy, normal vWF-CP was recovered in plasma from 2 acquired TTP patients. Among BMT patients, plasma from 4 BMT-TMA showed normal vWF-CP activities as 55-111%, whereas plasma from 2 BMT-VOD revealed low vWF-CP activity, as 24% and 37%, respectively. Thus, measurement of vWF-CP is crucial to predict differentiation of primary forms of TMA to establish the pathogenesis in varied endothelial dysfunction. PMID- 12884738 TI - [Intraoperative evaluation of axillary lymph nodes for micrometastases using immunohistochemistry--preliminary study]. AB - The increase of utilization of sentinel lymph nodes concept for breast carcinoma has made intraoperative evaluation of immunohistochemistry using epithelial markers attractive. At present the optimal procedures for intraoperative detection of micrometastasis of axillary lymph nodes has not been established. The purpose of this study is to evaluate the immunohistochemistry for intraoperative diagnosis of axillary lymph nodes in patients with breast cancer. Lymph nodes from 170 patients(1048 lymph nodes) were examined immunohistochemistry using anti cytokeratin, compared with intraoperative frozen section of same lymph nodes with H & E staining. Tumor metastases were found in 50 patients(92 lymph nodes) in H & E staining section, compared with 64 patients(113 lymph nodes) stained with anti-cytokeratin. Of 14 patients whose metastases were detected by immunohistochemistry. Routine intraoperative frozen diagnosis using H & E stainings significantly underestimates lymph nodes metastases. The insufficient diagnosis may be overcome by immunohistochemistry using anti-cytokeratin and careful examination of routine sections with good qualities. The true clinical significance of these micrometastases will be determined by long term follow up studies. PMID- 12884739 TI - [Analyses of pathogenesis and therapeutic approaches for hereditary amyloidosis]. AB - Amyloidosis is a disorder of protein metabolism in which normally soluble autologous proteins are deposited in tissues as abnormal insoluble fibrils, causing structural and functional disruptions. We have recently identified the novel localized amyloidosis accompanied by trichiasis. The precursor protein of amyloid deposits was mutated lactoferrin and all the patients had lactoferrin Glu561Asp. The disease was classified hereditary amyloidosis whose risk factor is trichiasis. We examined the therapeutic possibilities for mutated transthyretin(ATTR) related familial amyloidotic polyneurpathy(FAP), one of the systemic amyloidoses. Cr3+ suppressed amyloid formation by stabilizing ATTR structure in vitro. BSB is a useful new diagnostic tool to detect amyloid deposits both in in vitro and in vivo and may have therapeutic potential for preventing amyloid deposition. Gene therapy using single-stranded oligonuclotides(SSOs) may become a promising tool for therapy instead of liver transplantation. SSOs with athrocollagen effectively replaced the TTR gene both in vitro and in vivo. PMID- 12884740 TI - [Molecular mechanisms of hereditary neuropathy: genotype-phenotype correlation]. AB - Hereditary neuropathies are classified into several subtypes according to clinical, electrophysiologic and pathologic findings. Recent genetic studies have revealed their phenotypic and genetic diversities. In the primary peripheral demyelinating neuropathies(CMT1), at least 9 genes have been associated with the disorders; altered dosage of peripheral myelin protein 22(PMP22) or point mutation of PMP22, the gap junction protein 1(GJB1), the myelin protein zero gene(MPZ), the early growth response gene 2(EGR2), the myotubularin-related protein 2 gene(MTMR2), the N-myc downstream-regulated gene 1 (NDRG1), the L periaxin gene(PRX), SRY-related HMG-BOX gene 10(SOX10) and the ganglioside induced differentiation-associated protein 1 gene(GDAP1). In the primary peripheral axonal neuropathies(CMT2), at least 8 genes have been associated with these disorders; the neurofilament light chain gene(NEFL), the kinesin 1B gene(KIF1B), the gigaxonin gene(GAN1), Lamin A/C(LMNA) and tyrosyl-DNA phosphodiesterase 1(TDP1). In addition, some mutations in GJB1, MPZ and GDAP1 also present with clinical and electrophysiologic findings of CMT2. Mutation of NEFL or KIF1B cause dominantly inherited axonal neuropathies, whereas mutation of GJB1 or MPZ can present as genocopies of dominant axonal neuropathies. In addition to the above diseases, we have reported a new type of NMSNP(MIM # *604484) characterized by proximal dominant neurogenic atrophy, obvious sensory nerve involvement and the gene locus on 3q13. Here, we summarize the genetic bases of hereditary neuropathies and attempt to highlight significant genotype phenotype correlations. PMID- 12884741 TI - [Genetic diagnosis of diabetes mellitus: Wolfram syndrome--from positional cloning to DNA diagnosis]. AB - Wolfram syndrome(WFS, DIDMOAD syndrome) is an autosomal recessive neurodegenerative disorder characterized by juvenile-onset, insulin-dependent diabetes mellitus and optic atrophy, often accompanied by other symptoms including diabetes insipidus, neurosensory deafness, urinary tract and neurological abnormalities. In patients, beta-cells are selectively lost from their pancreatic islets of Langerhans. By positional cloning, a novel gene named WFS1 was identified and many mutations were subsequently identified in patients with WFS. Most of the mutations were identified in the largest 8th exon, however, they also existed in other exons. No common mutations were found and founder effect was not observed except in Spanish families. The WFS1 gene encodes a putative transmembrane protein of 100.3 kDa localized in endoplasmic reticulum. More recently, WFS1 mutations were also identified in patients with dominantly inherited low-frequency sensorineural hearing loss. Clarification of the functions of the WFS1 protein, as well as the phenotype-genotype relationship, will help improve understanding of the pathophysiology of diseases caused by the WFS1 gene. PMID- 12884742 TI - [Gene diagnosis of hemoglobinopathies--the determination of breakpoints for a large deletion]. AB - A large number of mutations leading to hemoglobinopathies(abnormal hemoglobins, thalassemias) have been discovered. Gene diagnosis for a point mutation or a deletion/insertion of a few nucleotides is now readily performed. For a large deletion, once the precise breakpoints are unraveled, the same type of deletion is promptly diagnosed by gap PCR. However, a number of new types of large deletions remain unexamined. They need meticulous Southern blot analysis and/or cloning. Here we present a new technique to determine their precise breakpoints. One of the two breakpoints needs to be first assigned within the 5-kb portion estimated by gene dosage by quantitative PCR. The other breakpoint is left unexamined. The genome DNA is digested with one of eight kinds of endonucleases and subjected to recombination with pUC18 cloning vector digested with the same endonuclease. The gap PCR is subsequently performed between the common primers of pUC18 and five arbitrary primers within the aforementioned 5-kb portion. An abnormal gap PCR product, if detected by electrophoresis, discloses precise 5' and 3' breakpoints after direct sequencing. This method successfully disclosed the breakpoints for two epsilon gamma delta beta-thalassemias, one delta beta thalassemia and one beta-thalassmeia in a relatively short period. All are new mutations. This method uses neither cloning procedures nor Southern blot, but employs gene dosage estimation and PCR. Thus, it is relatively simple. The progress of the genome project facilitated analysis of any large deletions. PMID- 12884743 TI - [Management of laboratory for clinical physiology]. AB - Physiological examination has the following characteristics: 1. Patient understanding and cooperation are required. 2. Accuracy of the results of examination is influenced by technician skill and experience. 3. Accidents may occur during examination. 4. Examination and judgment of the results proceed under cooperation between doctors and technicians. 5. Examination can be performed at patient's bedside. Recently, laboratories for clinical physiology have been required to provide high-quality examinations with an efficient cost performance. They must not only integrate and conduct various physiological tests on demand, but also arrange the proper number of staff for the proper number of instruments. It is important to reinforce the risk management against unexpected accidents and to allocate a chief technician for flexible management of the laboratory. Equipment should be purchased according to systematic planning, novel technology should be actively introduced into laboratory medicine, and excellent technicians to handle it should be brought up. Standardization of the test methods and the quality control are necessary to guarantee the quality of the test results. These are issues to be solved by the society of laboratory medicine and not by a single institution. PMID- 12884744 TI - [Current neurophysiological tests and revised JSCN technical standards for clinical EEG]. AB - The purpose of this lecture is to review the development of current neurophysiology and the revised standard of society for clinical EEG. 1. The improvement of neurophysiological tests. 1) EEG and evoked potential: EEG and evoked potential testing includes the routine EEG recording, EEG monitoring in surgical operation, all night sleep polygraph for the diagnosis of sleep apnea syndrome and many kinds of brain evoked potentials. Especially, the P300 component in the ERP(event-related evoked potential) is useful for the testing of essential brain functions. 2) EMG and evoked EMG: These tests are applied for the diagnosis of neurogenic, myogenic and neuromuscular junction disorder, and also the single fiber EMG using micro needle electrode is useful for the diagnosis of myasthenia gravis. Motor and sensory nerve conduction velocity are calculated from the latency of evoked EMGs. Furthermore, the distribution of these conduction velocities in many nerve fibers is measured by the collision technique. 3) Other tests: Near-infrared spectroscopy for the testing of brain functions has made rapid progress, and the transcranial magnetic stimulation method has come to be used for evaluation of functional diseases in the pyramidal tract, cerebellum and the spinal cord. 2. The revised JSCN technical standards for clinical EEG. The revised recording conditions of ECI(electro cerebral inactivity: flat EEG) in brain death are the focus of this lecture. PMID- 12884745 TI - [Pulmonary function tests]. AB - The spirometry has been most valuable pulmonary function test and it defines pulmonary physiology. But the spirometry has not been widely used by general physicians in Japan. The spirometry is effort-dependent test, so, they seem to keep it at a distance. It's desirable that pulmonary function tests are effort independent. We introduce some effort-independent pulmonary function tests and refer to analysis of exhaled breath condensate. PMID- 12884746 TI - [Guideline for hyperlipidemia]. AB - The guideline for hyperlipidemia was first discussed at the meeting of the Japan Atherosclerosis Society in 1987 and further discussed and published for the first time by the Society in 1997. Thereafter, a new guideline "Japan Atherosclerosis Society(JAS) Guidelines for Diagnosis and Treatment of Atherosclerotic Cardiovascular Diseases" was released in September 2002 taking the results of J LIT study and others into consideration. The diagnostic values of TC, LDL-C, HDL C and TG for hyperlipidemia are > or = 220 mg/dl, > or = 140 mg/dl, < 40 mg/dl, and > or = 150 mg/dl, respectively, the same figures as those of a previous guideline in 1997. Other criteria were made as management goals for patients with coronary heart disease in each category, classified by the number of risk factors. According to the criteria, the recommended level of TC is < 240 mg/dl in males aged less than 45 with no risk factors. If any result is abnormal, treatment by life style and dietary modification should be started first and drug therapy should be a second choice. It should be noted that this guideline helps to inform, not replace, the physician's clinical judgement, which must ultimately determine the appropriate treatment for each individual. PMID- 12884747 TI - [Guidelines for the management of hypertension in Japan]. AB - Hypertension is one of the most significant risk factors for cerebrovascular and heart diseases, which rank as the second and third most frequent causes of death in Japan. Two guidelines, Guidelines for the Management of Hypertension for General Practitioners(JSH2000) and Guideline for Treatment of Hypertension in the Elderly(2002 Revised Version), are currently available for treatment of hypertension in Japan. Both guidelines include indications for the treatment of hypertension, classification and evaluation of blood pressure(diagnosis), treatment strategy, modification of lifestyle and treatment with antihypertensive drugs. The guidelines are coming into clinical practice as useful references for the control and treatment of hypertension in Japan. They have advantages based on evidence from many recent studies in Japan and on consideration of the unique life style of Japanese, since the treatment guidelines for hypertension previously referenced are essentially guidelines for individuals in Western countries, primarily Caucasians. In Guideline for Treatment of Hypertension in the Elderly(2002 Revised Version), the opinions and criticisms of the Japanese hypertension experts are also incorporated. In future research, there is a need for further observational epidemiological studies and intervention trials in Japanese patients to provide direct evidence about the risks associated with blood pressure and other risk factors, as well as the antihypertensive drug effects. PMID- 12884748 TI - [Diabetes mellitus and HbA1c]. AB - Since the Japan Diabetes Association(JDS) began to distribute Lot 1 as the primary calibrator for measuring Hemoglobin A1c(HbA1c) to manufactures of reagents and devices in 1995, the standardization of the measurement of HbA1c has been achieved throughout Japan. In 1999, JDS proposed a new diagnostic criterion for diabetes mellitus, and HbA1c was adopted firstly as an assistant index for diagnosis of diabetes mellitus, after HbA1c measurement was standardized by JDS. Furthermore, JDS published a clinical guideline for treatment of diabetes mellitus in 1999, in which it was stated that HbA1c below 6.5% should be one of the goals. This statement was made based on the results of the Kumamoto Study. Because Lot 1 was made mainly in order to minimize the difference among measurements by HPLC devices, Lot 1 is a freeze-dried material. Since some faults of Lot 1 were pointed out after its distribution, in 2001 JDS approved Lot 2 as the new standard material, a deeply frozen material. Currently in Japan, the standardization of HbA1c is maintained with Lot 2. PMID- 12884749 TI - [The guidelines for the diagnosis and monitoring of endocrine diseases(except of diabetes mellitus)]. AB - The 6th draft for the guidelines for the diagnosis of thyroid disease has been published from the working group of Japan Endocrine Society in 2002. However, there are still controversial about guidelines for diagnosis of Graves disease. Serum-based tests for measuring the concentration of the TRAb and TSAb are available. We should use these tests rather than measuring of radioisotope uptake recommended in the 6th draft. PMID- 12884750 TI - [Involvement of unfolded protein responses in neurodegeneration]. AB - Various stresses cause the accumulation of unfolded proteins in the endoplasmic reticulum (ER). To manage the state, cells have the unfolded protein responses (UPR). If the UPR is unsuccessful, ER-mediated apoptosis occurs. To date, three types of UPR, i.e. the induction of chaperones, the translation block, and ER associated degradation (ERAD) have been reported. To sense the accumulation of unfolded proteins, the ER has IRE1, PERK, and ATF6. The pathways mediated by IRE1 and ATF6 cause the induction of chaperones. The pathway mediated by PERK causes a translation block. The induction of caspase 12, the activation of the JNK pathway, and the induction of CHOP have been reported as apoptosis caused by ER stress. The stability of the cell is based on the balance between UPR and ER mediated apoptosis. Recently several diseases have been reported to be related to ER stress. We reported that mutant presenilin 1 causes a vulnerability to ER stress because it attenuates the activation of IRE1, PERK, and ATF6. Recent reports have also shown that Parkinson disease and polyglutamine diseases are relevant to ER stress. Therefore it is suggested that the ER stress story is the common mechanism for neurodegerative disorders. PMID- 12884751 TI - [New aspects of neuroprotective effects of nonsteroidal anti-inflammatory drugs]. AB - Nonsteroidal anti-inflammatory drugs (NSAIDs) exert anti-inflammatory, analgesic and antipyretic activities and are involved in the suppression of prostaglandin synthesis by inhibiting cyclooxygenase (COX), a prostaglandin synthesizing enzyme. It has been recently revealed that NSAIDs also possess inhibitory effects on the generating system of nitric oxide radicals and modulating effects on transcription factors and nuclear receptors which are related to inflammatory reactions. Since it has been reported that inflammatory processes are associated with the pathophysiology of several neurodegenerative diseases and that NSAIDs inhibit amyloid beta-protein-induced neurotoxicity to reduce the risk for Alzheimer's disease, a number of studies have been conducted focusing on the neuroprotective effects of NSAIDs. It has been clarified that the drugs exert neuroprotective effects, which are not related to their COX-inhibiting property, on pathophysiology of various neurological disorders. In this article, new aspects of neuroprotective effects of NSAIDs have been reviewed, especially, in Alzheimer's disease and Parkinson's disease, discussing various pharmacological effects of NSAIDs other than their inhibitory action on COX. PMID- 12884752 TI - [Direct involvement of the supraspinal phosphoinositide 3-kinase/phospholipase C gamma 1 pathway in the mu-opioid receptor agonist-induced supraspinal antinociception in the mouse]. AB - Recent works have documented that the stimulation of the mu-opioid receptor (MOR) can activate phosphoinositide-specific phospholipase C (PI-PLC). Here I demonstrate that PLC beta 3 isoform activated by beta gamma subunit of G-protein (G beta gamma) in the brain may contribute to the negative modulation for supraspinal antinociception induced by morphine in mice. In immunohistochemical studies, phosphoinositide 3-kinase (PI3K) was detected in the membrane of the cell soma and the immunoreactivity of PI3K (PI3K-IR) was almost overlapped with MOR-IR and PLC gamma 1-IR in the periaqueductal gray matter (PAG) that is considered to be one of the most important sites for the expression of MOR mediated antinociception. Morphine produced a marked increase in the protein level of membrane-bound PLC gamma 1, and this increase induced by morphine was significantly inhibited by intracerebroventricullar (i.c.v.) pretreatment with PI3K inhibitors at the dosage that suppressed the morphine-induced supraspinal antinociception. Furthermore, morphine also caused a robust increase in the number of phosphorylated-PLC gamma 1 (p-PLC gamma 1) expressing cells in the PAG. It is worthwhile to note that MOR-IR was overlapped with p-PLC gamma 1-IR in the same cells that also contained PI3K in this region. Based on these findings, the next experiment was designed to investigate whether a deletion of the PLC gamma 1 gene by i.c.v. pretreatment with antisense oligodeoxynucleotide (AS-ODN) against PLC gamma 1 could affect the antinociception induced by MOR agonists. Pretreatment with AS-ODN against PLC gamma 1 revealed a significant inhibition of supraspinal antinociception induced by MOR-agonists. In addition, the morphine induced supraspinal antinociception was suppressed by the blockade of the G beta gamma subunit that can directly activate both PI3K and PLC gamma 1. Moreover, mice lacking the gene for inositol 1,4,5-trisphosphate (IP3)-sensitive receptors, which can modulate the release of Ca2+ from the endoplasmic reticulum, exhibited a significant inhibition of the morphine-induced antinociception. Collectively, these findings raise the possibility that the activation of the PLC pathway associated with the stimulation of PI3K and/or G beta gamma is implicated in supraspinal antinociception induced by MOR agonists in mice. PMID- 12884753 TI - [Regulation of development of oligodendrocyte]. AB - Oligodendrocyte (OL) progenitors in the mouse spinal cord are generated from a strictly restricted region in the ventricular zone of the ventral cord as early as on embryonic day 11 (E11). We previously reported that one of the factors that restrict this ventral specific appearance of OLs is an inhibitory factor secreted from the dorsal spinal cord, in addition to well-known stimulatory ventral factors such as sonic hedgehog. We characterized the developmental change of the inhibitory activities. They were very strong at E11, gradually reduced, and disappeared by E14. This pattern seemed to be well correlated with the developmental profile of Wnt3a expression at/near the roof plate. A conditioned medium of L cells that stably express Wnt3a showed significant reduction of O4 positive OLs in the ventral spinal cord explants, indicating that Wnt3a is one of the dorsal factors that inhibit OL development. Addition of Wnt3a supernatant to CG4 cells, an OL progenitor strain, and to the dissociated primary cultured cells suggested that Wnt3a directly acts on OL lineage cells and inhibits a differentiation step from OL progenitor to O4-positive stage. Thus, Wnt3a may directly control the timing of OL differentiation and the motility of OL lineage cells. A population of myelinating OLS in the dorsal area of telencephalon was further demonstrated to be ventral origin by the newly established cell marking system using in utero DNA electroporation. PMID- 12884754 TI - [Molecular dynamics of K+ transport and its crucial involvement in signal transduction]. AB - Potassium ion (K+) is the only cation whose concentration in intracellular solution is more abundant than in extracellular fluid. In higher animals, each of the cells, which is bathed in the extracellular fluid containing approximately 5 mM K+, has approximately 140 mM K+ in its cytoplasm. Physiological experiments have reported that the glial cells such as brain astrocytes and retinal Muller cells highly express K+ conductance on their membrane. The K+ conductance is thought to be responsible for transport of K+ from the extracellular regions of high K+, which is elicited by the excitation of neurons, to those of low K+. This so-called glial K(+)-buffering action is essential for maintenance of neuronal activity. Types and distribution of the K+ channels involved in the buffering action, however, have remained unknown for a long time. Recently, we have identified that an inwardly rectifying K+ channel, Kir4.1, is dominated in glial cells and plays a central role in the K(+)-buffering action. We also found that Kir4.1 is enriched on the processes of astrocytes wrapping synapses and blood vessels. Furthermore, some of the Kir4.1 is assembled with Kir5.1, suggesting that a heteromultimer of Kir4.1 and Kir5.1 plays a distinct role in the glia. PMID- 12884755 TI - [Molecular mechanisms of nociception]. AB - Capsaicin, the main ingredient in 'hot' chili peppers, elicits burning pain by activating nociceptors. The cloned capsaicin receptor (TRPV1) is a nonselective cation channel with six transmembrane domains, and is activated not only by capsaicin but also by noxious heat (> 43 degrees C) or protons (acidification), both of which cause pain in vivo. Furthermore, analyses of mice lacking VR1 showed that VR1 is essential for selective modalities of pain sensation and for tissue injury-induced thermal hyperalgesia. Tissue damage produces an array of chemical mediators that activate or sensitize nociceptor terminals to elicit pain. Important components of this pro-algesic response are ATP and bradykinin. In cells expressing TRPV1, ATP or bradykinin increased the currents evoked by capsaicin or protons through activation of metabotropic P2Y or B2 bradykinin receptors. In the presence of ATP or bradykinin, the temperature threshold for VR1 activation was reduced from 42 degrees C to 30-35 degrees C, such that normally non-painful normal body temperatures were capable of activating TRPV1, thereby leading to the sensation of pain. Direct phosphorylation of TRPV1 by PKC epsilon was confirmed and the involved two serine residues were determined. This represents a novel mechanism through which ATP or bradykinin in response to tissue trauma might trigger the sensation of pain. PMID- 12884756 TI - [Neural and molecular mechanisms of fatigue and recovery from fatigue]. AB - Fatigue is an indispensable biosignal for maintaining life. However, the neural/molecular mechanisms of fatigue are still unclear. Here, the recent progress in this field is introduced, mostly through our project research under the control of the Ministry of Education, Culture, Sports, Science and Technology (MEXT), Japanese Government. The sensing mechanisms of fatigue in the brain might be related to the orbitofrontal-dorsoprefrontal-cingulate triangle circuits. We hypothesized the neural circuits for fatigue sensation through our PET study on chronic fatigue patients and healthy volunteers. The serotonergic system might be involved in the sensation in the prefrontal cortex, although the hyperserotonin hypothesis might be wrong. The fatigue sensation is somehow related to hypofunction of the glutamatergic system through reduced uptake of acetyl carnitine in the discrete brain regions. For creation of new methods and drugs overcoming fatigue, different types of animal models of fatigue were developed. By using such animal models, the green leaf odor, ascorbic acid, acetyl carnitine, and tetrahydrobiopterin have been found to be effective. PMID- 12884757 TI - [Home parenteral and enteral nutritional in Japan-present and future]. PMID- 12884758 TI - [New diagnostic method of gastrointestinal disease with CT and MRI, especially three-dimensional imaging]. PMID- 12884759 TI - [Three-dimensional endoscopic ultrasonography]. PMID- 12884760 TI - [Clinical pathway for inpatients with gastric ulcer: evaluation of usefulness]. AB - Based on the results of a retrospective review of clinical data on inpatients with gastric ulcer treated at our department, we devised on original clinical pathway and tested it in the clinical setting. From the results obtained, we created an improved clinical pathway and evaluated its usefulness. The duration of hospitalization was 16.2 +/- 6.9 (mean +/- SD) days in the non-path group, 14.1 +/- 3.0 days in the original path group, and 10.9 +/- 2.0 days in the improved path group. The hospital time was significantly shorter in the improved path group. For patients with bleeding gastric ulcer, the duration of hospitalization was 18.0 +/- 6.3 days in the non-path group, 15.1 +/- 2.3 days in the original path group, and 11.2 +/- 1.8 days in the improved path group. This period was also significantly shorter in the improved path group. With regard to the occurrence of rebleeding from the gastric ulcers, there were no significant differences between the non-path group and both clinical path groups. These results indicate that devising a clinical pathway is useful for shortening the duration of hospitalization for patients with gastric ulcer. PMID- 12884761 TI - [Two cases of Sister Mary Joseph's nodule due to colon cancer]. PMID- 12884762 TI - [A case of Crohn's disease complicated with hepatocellular carcinoma]. PMID- 12884763 TI - [A case of primary small intestinal gastrointestinal stromal tumor: an intraperitoneal bleeding from greater omentum metastasis caused by administration of imatinib mesylate]. PMID- 12884764 TI - [A case of angiosarcoma metastatic to the small intestine]. PMID- 12884765 TI - [Percutaneous radiofrequency ablation of bone metastasis to the rib from hepatocellular carcinoma: a case report]. PMID- 12884766 TI - [A case of intraductal papillary-mucinous carcinoma without mucin-hypersecretion of the pancreas]. PMID- 12884767 TI - [A case of groove pancreatitis pure type treated with conservative therapy]. PMID- 12884768 TI - [A case of an anomalous junction of the pancreatico-biliary ductal system with intraductal papillary-mucinous carcinoma of the common canal of pancreas]. PMID- 12884769 TI - [A case of pancreatic carcinoid tumor diagnosed 10 years after operation of primary hepatic carcinoid tumor]. PMID- 12884770 TI - [A case of acute pancreatitis induced by metastasis of renal cell carcinoma]. PMID- 12884772 TI - [Antiinfective host defense mechanism: toll-like receptors and innate immunity]. AB - The innate immune system has evolved as the first line of defense against invading microorganisms. The recent discovery of the toll-like receptors(TLRs) has rapidly expanded our knowledge of molecular events that initiate host pathogen interactions. The TLRs, which are expressed on the surface of cells, involved in innate immune recognition, including macrophages and dendritic cells, have a crucial role in the detection of microbial infection. Signals initiated by the interaction of TLRs with pathogen-associated molecular patterns (PAMPs) induce activation of the inflammatory and antimicrobial innate immune response. Ten members of the TLR family have been identified, and they appear to recognize PAMPs, including lipopolysaccharide, peptidoglycan, and bacterial DNA. There has been considerable interest in how adaptive immune responses are controlled by the innate immune system. Recent studies have suggested that TLRs may control the induction of Th1 responses and that a separate system of recognition regulates the Th2 response. Thus TLR signaling represents a key component in the innate immune response to microbial infection. PMID- 12884773 TI - [The pathogenesis of the systemic inflammatory response syndrome and compensatory antiinflammatory response syndrome following surgical stress]. AB - The inflammation charge cells such as the macrophages, neutrophils, endothelial cells, and fibroblasts are activated in surgical sites when tissue injury occurs due to the operation. Proinflammatory cytokines such as tumor necrosis factor alpha, interleukin (IL)-1, IL-6, and IL-8 are induced from the activated inflammation charge cells. These cytokines amplify the information by autocrine and paracrine action, induce the production of other cytokines, and send the information to the whole body. Increases in body temperature, pulse rate, and leukocyte counts are then observed. This condition is called the systemic inflammatory response syndrome (SIRS) clinically. On the other hand, the production of antiinflammatory cytokines against SIRS is induced simultaneously. The condition in which antiinflammatory cytokines are produced in excess and become dominant systemically is called the compensatory antiinflammatory response syndrome (CARS). No standard concrete diagnosis for CARS has been established, although CARS is a pathogenetic concept. However, recently the analysis of cytokine production and various types of surface molecules in the inflammation charge cells became possible with the development of molecular biological methods. As a result, it was found that the immune system is controlled by the balance between proinflammatory and antiinflammatory cytokine production, and if the compensatory antiinflammatory reaction is sufficiently severe, it will manifest clinically as anergy, with increased susceptibility to infection. In this manuscript, we explain the pathogenesis of SIRS and CARS following surgical stress by analyzing cytokine production and surface membrane molecules in the inflammation charge cells. PMID- 12884774 TI - [Role of polymorphonuclear neutrophils and macrophages in the prevention of postoperative infections]. AB - Polymorphonuclear neutrophils (PMNs) and macrophages are recognized to serve as the first line of defense against bacterial contamination during the perioperative period. Chemoattractants produced by macrophages cause PMN accumulation at the inflammatory site. Proinflammatory cytokines released by macrophages increase adhesion molecule expression on the surface of endothelial cells. P- and E-selectins produce leukocyte rolling, whereas beta 2-integrin-ICAM 1 interaction causes firm adhesion of leukocytes to the endothelium, followed by migration to the interstitium. Thus appropriate activation of the inflammatory cascade leads to leukocyte migration to the infectious focus. In particular, during the first several hours after the onset of bacterial contamination, massive exudation of PMNs is generally observed. PMNs and macrophages phagocytose the bacteria via opsonization and kill them with oxygen products or proteolytic enzymes. Malnutrition and the lack of enteral nutrition are assumed to impair PMN macrophage-dependent host defense by derangement of adhesion molecule expression and the cytokine milieu. Recently, sticky PMNs before surgery have been reported to increase infectious morbidity after surgery. In addition, augmented alpha 4 integrin-VCAM-1 interaction has been demonstrated during sepsis, which is possibly a mechanism for increased PMN adhesion and resultant organ injury. Further study is needed to clarify the mechanisms of the disturbed function of PMNs and macrophages under various conditions. PMID- 12884775 TI - [Humoral mediators in the pathophysiology of organ dysfunction in sepsis]. AB - It is recognized that various humoral mediators, especially inflammatory cytokines such as tumor necrosis-factor-alpha and interleukin (IL)-1 beta, play a key role in the pathophysiology of septic shock and organ dysfunction following severe infection. We have recently begun to employ a rapid measurement system that allows us to measure blood IL-6 levels within 30 min using a chemiluminescent enzyme immunoassay. IL-6 blood levels reflect well the activation of the cytokine cascade and correlated well with the severity of patient's, condition. The ratio of simultaneously measured IL-6 blood levels in peripheral and pulmonary arteries is useful to identify the site responsible for cytokine production. Early initiation of PMMA-CHDF as a cytokine modulator of septic shock results in improved recovery from shock and in improved outcome. Genetic analysis of cytokine-related genes revealed a high frequency of a specific genotype in patients who have extremely high IL-6 blood levels. Screening of patients at high risk of hypercytokinemia with the specific genotype would enable appropriate treatments and contribute to future tailor-made medicine. PMID- 12884776 TI - [Molecular diagnosis of infectious disease]. AB - With the development of molecular biology over the decades, molecular technologies have become available for clinical diagnosis of infectious disease. The principle of the technique is to detect specific genes of microorganisms. Compared with the traditional culturing techniques, molecular technologies have several advantages: 1) direct detection and rapid identification of organisms that are slow growing or those currently lacking a system for in vitro cultivation; 2) nucleic acid-based methods for epidemiologic typing of microorganisms; and 3) identification of genotypic markers of microbial resistance to specific antibiotics. When sufficient amounts of nucleic acid can be obtained, direct probe tests (simple molecular hybridization-based techniques) may be performed. In many instances with small amounts of sample nucleic acids, amplification techniques(e.g., polymerase chain reaction[PCR]) are extremely useful. However, the application of PCR to clinical specimens has potential pitfalls due to the susceptibility of PCR to inhibitors, contamination, and experimental conditions. In some cases, DNA fingerprinting or restriction fragment-length polymorphism analysis is used and it has become the method of choice for epidemiologic analysis. The major disadvantage of the molecular method is that we cannot differentiate whether the microorganisms identified are alive or dead and chemosensitivity testing cannot be performed. PMID- 12884777 TI - [The application of molecular biology to anti-endotoxin therapies]. AB - Toll-like receptors (TLRs), recently identified on macrophages and dendritic cells in mammals, recognize a specific pattern of pathogen components, including endotoxins(lipopolysaccharide). Pathogen recognition by TLRs activates the innate immune system through the signaling pathway and provokes inflammatory responses, such as inducing the production of cytokines. Therefore the specific inhibition of the signaling pathway and the administration of excess inflammatory responses have useful potential in the management of sepsis syndrome. Currently, several monoclonal antibodies are applicable to the treatment of autoimmune diseases and cancer. On the other hand, immunotherapies against proinflammatory cytokines in septic shock have failed to demonstrate clinical benefit. In this review, we summarize recent views of novel therapeutic targets, provided from molecular biologic studies of gram-negative infection. PMID- 12884778 TI - [Microbiological approach to antibiotic therapy against multidrug-resistant bacteria]. AB - Many severe infections that occur after gastrointestinal surgery are caused by multidrug-resistant bacteria such as methicillin-resistant staphylococcus aureus and Pseudomonas aeruginosa. (P. aeruginosa,) which has been isolated from such cases, is resistant to many beta-lactamantibiotics, including imipenem (IPM). IPM resistant P. aeruginosa lacks the outer membrane proteins (Opr) D, OprD2, OprC, and OprE1, which together normally form an opening in the outer membrane. Therefore IPM cannot pass through the outer membrane. But it is also known that meropenem, caftazidime, and ciprofloxacin are not restricted by this effect and that they can easily pass through the outer membrane. Therefore if we can identify these genes by using the polymerase chain reaction, appropriate antibiotics can be swiftly administered without waiting for the results of drug sensitivity tests. PMID- 12884779 TI - [Medical information service of clinical practice guidelines]. PMID- 12884780 TI - [What we can learn from a case of medical malpractice]. PMID- 12884781 TI - [Quantitative study of 99mTc-technegas SPECT for ventilatory impairment in pulmonary emphysema--regional distribution: correlation of SPECT with pulmonary function test]. AB - 99mTc-Technegas scintigraphy is used for evaluation of abnormality of ventilation in pulmonary emphysema. Although the abnormality of ventilation distribution is very easy to find, there is not an objective index. The evaluation is subjective and different by each radiologist. It was also difficult to compare each cases and the clinical course in the same case. The present study for quantitative evaluation has proved that excellent correlations is obtained between the mean voxel values of the lung and stage classification. Furthermore, a correlation is observed between the mean and FEV1.0%. These deta indicate that quantitative analysis of SPECT seem to be useful to classify the clinical stage and compare the each cases. And respiratory function decreases as much as chronic pulmonary emphysema exists in the lower lung field. PMID- 12884782 TI - [The development of new method for the assessment of perfusion reserve using split dose iodine-123-IMP SPECT: one-day protocol by modified ARG method]. AB - Evaluation of the regional cerebrovascular reactivity (rCVR) to a cerebral vasodilatory stimulus is important in the investigation of patients with ischemic cerebrovascular disease. We devised a simplified one-day protocol technique using [123I]N-isopropyl-p-iodoamphetamine (IMP) autoradiography (ARG) with SPECT. To validate the accuracy of IMP-ARG for quantifying rCVR to acetazolamide, we compared rCVR measured using IMP-ARG with rCVR calculated using IMP split dose method of microsphere model. Twenty patients with chronic steno-occlusive disease in a unilateral major cerebral artery underwent 123I-SPECT. On rCBF SPECT image above 3.5 cm from OM line, large cortical regions of interest (ROI) was bilaterally determined for bilateral middle cerebral artery and anterior cerebral artery. Based on rCBF values in each ROI, rCVR to acetazolamide was calculated. Significant correlation was observed between rCVR values obtained using IMP-ARG and microsphere model IMP methods in the 80 ROIs examined in the 20 patients (r = 0.72; p < 0.001). The result demonstrated that [123I]IMP-ARG split dose method can quantify rCVR non-invasively in a short time. PMID- 12884783 TI - [Evaluation of the 123I-IMP Patlak plot method using the pulmonary differential curve as an input function: a comparison with cerebral blood flow (CBF) determined by the noninvasive micro-sphere (NIMS) method]. AB - We explored the possibility of applying the Patlak plot method to clinical practice as a simple non-invasive quantitative method of measuring cerebral blood flow using N-isopropyl-4-[123I]iodoamphetamine (123I-IMP). On the assumption that after temporarily accumulating in the lungs, all the administered 123I-IMP is eliminated by the pulmonary arterial flow for systemic diffusion, we collected dynamic data by setting an area ranging from the brain to the whole lung field within the field of the camera. The lung clearance curve L(t) was differentiated and divided by cardiac output. It was then converted a positive number by multiplying it of--1 to determine the volume of 123I-IMP tracer diffused in arterial blood per unit of time. The calculated concentration was defined as the arterial time activity curve A(t). A Patlak plot analysis was conducted between A(t) and the brain time activity curve B(t) to determine K1 (total cerebral blood flow [tCBF], ml/min) and Vn (nonspecific initial distribution volume, ml). The total volume of tracer diffused in the central cardiovascular system within a given (T) was also obtainer from the volume of tracer remaining in the lungs [Lpeak--L(T)], and by reporting this calculation over time, an accumulation curve was produced. By differentiating the obtained accumulation curve, we were able to estimate the volume of tracer diffused in the central cardiovascular system per unit of time. With this value used as the input function index I(t), a Patlak plot analysis was conducted to determine the unidirection influx index ki, which was then multiplied by 100 to obtain the brain perfusion index (IMP-BPI). The noninvasive micro-sphere method was performed concurrently on 16 patients with cerebrovascular and/or neurological disorders to obtain mean cerebral blood flow (mCBF). Correlations between K1 and between IMP-BPI and mCBF were then determined and compared. Both K1 and IMP-BPI obtained from ki were found to correlate highly with mCBF, r = 0.759 (y = 0.032x + 20.1) and r = 0.833 (y = 2.73x + 0.10) respectively, with a better result from IMP-BPI. These results indicate that the 123I-IMP Patlak plot method with a wide-field gamma camera is clinically applicable as a simpler noninvasive technique for measuring cerebral blood flow even when a simple input function is used. PMID- 12884784 TI - [Evaluation of lesions of malignant lymphoma using histograms of regional SUV]. AB - We designed new evaluation method using histogram of regional standardized uptake value (SUV) of every pixels of box shape volume of interest (VOI) on 18F-FDG PET. We evaluate lung and liver of normal volunteers and also the lesions of 4 cases of patients with non Hodgikin's malignant lymphoma using this method. SUV of pixels of pretreatment lesions shows log-normal distribution, whereas SUV of lung and liver show normal shape distribution. The next day of chemotherapy, 2 cases of them showed changes of distribution pattern from long-normal to normal with decrease of component of higher SUV values. No remarkable changes of distribution pattern were observed on other 2 cases, whereas decrease of regional mean values and max values of SUV were observed. And these findings were continued to 3 weeks later. These findings suggest that regional evaluation using histograms of SUV gives additional and predictive information for tumor therapies soon after the end of course. PMID- 12884785 TI - [Phase III additional clinical study of 111In-pentetreotide (MP-1727): diagnosis of gastrointestinal hormone producing tumors based on the presence of somatostatin receptors]. AB - Additional phase III multicenter clinical study was performed to investigate the efficacy, safety, and usefulness of somatostatin receptor scintigraphy using 111In-pentetreotide (MP-1727), which binds to somatostatin receptors. Forty patients were included in the study; Group A: 18 patients, gastrointestinal hormone producing tumors had been detected with conventional imaging modalities, Group B: 22 patients, no tumors had been detected with conventional imaging modalities in spite of high serum hormone levels. By comparing the results of the octreotide suppression test, 12/16 cases (75.0%) of Group A and 11/19 cases (57.9%) of Group B were assessed as "effective." By comparing the results of immunohistological examination, 5/9 cases (55.6%) of Group A and 2/4 cases (50.0%) of Group B were assessed as "effective." Severe adverse events were not observed in any of the evaluable 35 cases. MP-1727 was judged as clinically useful in 11/16 cases (68.8%) of Group A and 5/19 cases (26.3%) of group B. These results suggest that MP-1727 scintigraphy is very useful for the diagnosis and decision of the therapeutic strategy of gastrointestinal hormone producing tumors. PMID- 12884786 TI - [Current utilization of gated SPECT in Japan]. AB - The circumstances for gated SPECT in Japan were surveyed by the working group of the Japanese Society of Nuclear Medicine, using a questionnaire method. The questionnaire included the following items; 1) proportion of the gated SPECT, 2) indication, 3) radiopharmaceuticals, 4) study protocols, 5) unsuccessful cases, 6) data acquisition, 7) reconstruction methods, 8) software, and 9) reliable parameters. The questionnaires were sent to 345 institutions and were returned in 180 (52%), 177 of which used gated SPECT. Gated SPECT was mainly used for ischemic heart disease, exercise-rest or rest studies with 99mTc-myocardial agents were used in most of these patients. QGS software was used mostly for gated SPECT processing. Common parameters used in gated SPECT were ejection fraction, end-diastolic and systolic volume. The incidence of successful case for gated SPECT was 6.8% which patients had arrhythmia, large infarct, small heart, and high liver accumulation. PMID- 12884787 TI - [Evolution of chemotherapy for malignant brain tumors]. PMID- 12884788 TI - [Surgical treatment of temporal lobe epilepsy]. PMID- 12884789 TI - [Progressive brain injury]. AB - The aim of this study was to evaluate the clinical manifestations and prognostic factors of progressive brain injury following trauma. We reviewed the records of 779 patients with head injury who had an admission Glasgow Coma Scale of 9 or more; 70 (7.0%) developed progressive brain injury as evidenced on serial CT scans. Of these 70 patients, 19 (27.1%) had a subdural hematoma, 19 (27.1%) an epidural hematoma, 16 (22.9%) a cerebral contusion, 13 (18.6%) an intracerebral hematoma, and 3 (4.3%) a diffuse brain swelling. Three months after injury, 36 (51.4%) patients died, 2 (2.9%) were left in a vegetative state and 23 (32.9%) had a favorable outcome. The appearance of progressive brain injury was associated with patient age, admission Glasgow Coma Scale, injury mechanisms, skull fracture and hemorrhagic lesions on the initial CT scan. Patients with the extracerebral lesions deteriorated 4 hours after injury, whereas those with intracerebral lesions deteriorated 8 hours after injury. The outcome based on Glasgow Outcome Scale was significantly associated with age, type of intracranial lesion, Glasgow Coma Scale following deterioration, the mechanism of injury and surgical treatment. It is concluded that early repeated CT scan is indicated in patients with risk factors of developing progressive brain injury. PMID- 12884790 TI - Radiographical investigations of organic lesions of the hypothalamus in patients suffering from neurogenic pulmonary edema due to serious intracranial diseases: relationship between radiographical findings and outcome of patients suffering from neurogenic pulmonary edema. AB - Radiographical investigations of the hypothalamus by computerized tomography (CT) have rarely been performed despite the fact that the damage to the hypothalamus owing to serious intracranial organic diseases may cause neurogenic pulmonary edema (NPE). We presented 22 consecutive cases of patients suffering from NPE caused by serious intracranial organic diseases and investigated the relationship between NPE and abnormal radiographical findings of the hypothalamus. In 11 cases, organic lesions were noted in the hypothalami and 10 of these patients died of NPE (91.0%). In contrast, of the remaining 11 cases without significant radiographical findings of organic lesions in the hypothalami, only 2 patients were lost (18.2%). In general, various factors including systemic ones are considered to contribute to the prognosis of the patients suffering NPEs caused by serious intracranial diseases. It was concluded that hypothalamic damage was not always found by radiograph in patients with NPE due to critical intracranial diseases, but once abnormal findings in their hypothalamus of these patients were noted, their prognosis would become significantly poor (p < 0.05). PMID- 12884791 TI - [Therapeutic efficacy and prognostic factors in diffuse astrocytomas]. AB - Diffuse astrocytomas are slowly growing tumors with a relatively long overall survival. Considerable controversy exists as to the best therapeutic management for patients with such tumors. In the present study, we retrospectively analyzed a series of 64 patients with WHO grade II astrocytomas of the cerebral hemispheres. Gross total resection and interferon-beta therapy were significantly associated with both longer progression free survival (PFS) and overall survival (OS). Immediate postoperative radiation therapy did not prolong either the PFS or OS. The presence of promoter hypermethylation of the O6-methylguanine-DNA methyltransferase (MGMT) gene was an independent predictor of a shorter PFS. Our data suggest that radical surgery plus interferon-beta therapy may offer the best chance for long survival. Since the presence of MGMT methylation is a probable indication of an increased sensitivity to alkylating chemotherapeutic agents, determining the methylation status of MGMT could provide a potential basis for logical therapeutic intervention in identifying a subgroup of patients who could be candidates for early chemotherapy. PMID- 12884792 TI - [A case of metastatic suprasellar tumor with edema along the optic pathway]. AB - A rare case in presented of a patient with edema formation along the optic pathway, which was caused by metastatic suprasellar tumor mimicking craniopharyngioma. A 63-year-old female with visual disturbance, diabetes insipidus, and symptomatic panhypopituitarism was admitted to our hospital. CT images showed a suprasellar mass lesion. MR images demonstrated a heterogeneously enhanced tumor and edema along the optic nerve and tract. Although no cystic component is shown in radiological findings, these features are commonly observed in cases of craniopharyngioma. The patient underwent removal of the tumor by the transcranial approach. Since histological examination revealed adenocarcinoma, the tumor was diagnosed as metastatic brain tumor originating from non-recurrent rectal cancer operated on eight years previously. Conventional radiotherapy with a dose of 50GY was performed for the residual tumor. The tumor size was unchanged, but the edema had disappeared 7 months after the operation. Edema spreading along the optic tract was reported as a characteristic MR finding for diagnosis of craniopharyngiomas as distinguished from other common parasellar tumors. However, it is suggested that edema of the optic pathway seems to be caused not only by craniopharyngioma but also other suprasellar lesions. Even if there was a characteristic radiological finding for some specific diseases, metastasis should be kept in mind as a differential diagnosis, because of its various appearance in neuroimages. PMID- 12884793 TI - [Two cases of severe vertigo occasioned by neck rotation and its correlation with the direction of vertebral artery at its origin]. AB - We report two cases of severe vertigo with neck rotation. The first case is that of a 54-year-old woman. She suffered from severe vertigo during neck rotation to the right. Angiography demonstrated that the origin of the left vertebral artery (VA) became kinked when she turned her head to the right. The second case is that of a 59-year-old man. He suffered from severe vertigo during neck rotation to the left. Angiography demonstrated that the kink in the right VA at its origin became worse with the patient's turning his head to the left. We resected nerve fibers from the stellate ganglion at the origin of the respective vertebral arteries and corrected the kink in the arteries at their origin. Both patients were relieved of vertigo after the operation the fact that there was vertigo occasioned by neck rotation. The direction of neck rotation was identical with that of a kink in the VA at its origin in both patients. Therefore, the direction of neck rotation is useful information for the diagnosis of vertigo induced by neck rotation. In terms of the treatment, though vascular reconstruction such as vein graft bypass between the VA and the carotid artery or transposition of the VA onto the subclavian artery has been established for VA stenosis and occlusion, the resection of nerve fiber from the stellate ganglion at the origin of VA is also effective and safe, especially when a kink in the VA at its origin accompanies with neck rotation. PMID- 12884794 TI - [A dura-based intracranial schwannoma in the temporal fossa: a case report]. AB - We report a rare case of a dura-based intracranial schwannoma in the temporal fossa. A 77-year-old man admitted to our hospital with a 5 months history of mild recent memory disturbance. Magnetic resonance imaging (MRI) revealed a slight enhanced solid mass with a cystic mass in the right temporal lobe. Preoperative diagnosis was glioma and then total resection was performed by the transsylvian approach. Dense adhesion of the tumor to dura matter of the middle cranial fossa was confirmed during surgery. Microscopic examination of the tumor showed biphasic pattern with cellular Antoni A and hypocellular Antoni B areas. Immunohistochemically, the tumor was positive for S-100, vimentin and Schwann/2E and was negative for GFAP. From these findings, the tumor was diagnosed as a schwannoma arising from the dura matter of the middle fossa. The patient was discharged from our hospital with no neurological deficits. Intracranial schwannomas, which are not associated with cranial nerves, are rare. Our case was classified according to the origin into dura-based intracranial schwannoma. To our knowledge, only 14 cases of dura-based intracranial schwannoma have been reported, mostly in children and young adults. This represents the oldest case in the literature of dura-based intracranial schwannoma. PMID- 12884795 TI - [Surgical treatment of retroperitoneal presacral large schwannoma by the anterior transabdominal approach: two cases reports]. AB - Retroperitoneal schwannomas are relatively rare. However, when the tumors occur, they can be large and difficult to diagnose at an early stage because of their vague symptoms. Although surgery is the only treatment, the choice of surgical approach depends upon pathological type, the degree of sacral destruction, and intrapelvic extension. If a large component is present presacrally, an anterior transabdominal approach should be chosen to gain control of the tumor bleeding. We report two cases of large schwannoma in the retroperitoneal presacral space that was removed by the anterior transabdominal approach. We performed an intracapsular piecemeal resection with preservation of the tumor capsule, and were able to remove the tumor gross totally without any postoperative deficit. For this reason, we think that retroperitoneal presacral larger schwannoma should be removed leaving the capsule intact and using the anterior transabdominal approach to prevent injury of the adjacent structures. PMID- 12884796 TI - [Vertebrobasilar artery dissection with subarachnoid hemorrhage after brain stem infarct showing an improvement on angiography: case report]. AB - A 66-year-old male presented with dysarthria and right hemiparesis. Cerebral angiography at onset showed obliteration of the basilar artery and dilatation in the left vertebral artery. The patient's clinical symptoms were exaggerated and he was finally diagnosed to as suffering locked-in syndrome. Magnetic resonance imaging showed a brain stem infarction from the pons to the left middle cerebellar peduncle. We determined that the patient suffered dissection of the left vertebral artery at the basilar artery and treated, using the conservative therapy him of strict blood pressure control. A second angiography 13 days after onset showed recanalization of the basilar artery. At 29 days after onset, the patient fell into a coma with subarachnoid hemorrhage and acute hydrocephalus. Cerebral angiography revealed improvement in the irregularity and dilatation of the basilar artery, but the point of rupture could not be clearly identified. After performing proximal occlusion of the left vertebral artery by intravascular surgery, both right STA-SCA anastomosis and proximal occlusion of the right vertebral artery were carried out. Unfortunately, the patient died. Based on there data, it is appointed out that patients with a dissection of the vertebrobasilar artery must be followed up by serial angiography, and even if an angiographical improvement of the dissection is observed, the risk of subarachnoid hemorrhage still exists in patients suffering ischemic stroke. PMID- 12884797 TI - [Functional brain mapping detected by cortical stimulation using chronically implanted subdural electrodes: basic knowledge of clinical nerve physiology for neurosurgeons]. PMID- 12884798 TI - ["How I do it" no. 4, a case of mesenchymal chondrosarcoma in the upper thoracic spine]. PMID- 12884799 TI - [Hydrocephalus]. PMID- 12884800 TI - [Neuroendoscopic surgery for ventricular tumors]. PMID- 12884801 TI - [Endoscopic pituitary tumor surgery]. PMID- 12884802 TI - [Endoscopic evacuation of spontaneous intracerebral hemorrhage: a review]. PMID- 12884803 TI - [Temporal lobe epilepsy associated with cystic lesion in the temporal lobe and middle cranial fossa]. AB - Four patients out of 52 patients with temporal lobe epilepsy (TLE), who underwent epilepsy surgery in our hospital since September of 1994, had cystic lesions in the temporal lobe and middle cranial fossa. Case 1 had old hematoma cavity in the inferior temporal gyrus and chronic subdural electrode recording revealed the ictal onset zone to be localized in the ipsilateral medial temporal region. Case 2 had cystic ganglioglioma in the temporal tip, and intraoperative electrocorticography demonstrated independent paroxysmal activities from medial temporal region and temporal tip near the cyst. Both area were resected and the patients became seizure free. Case 3 and 4 had arachnoid cysts in the middle cranial fossa. Chronic subdural electrode recording revealed that the ictal onset zone was localized in the ipsilateral inferior temporal gyrus (that had microdysgenesis) in Case 3 and contralateral medial temporal region (that had hippocampal sclerosis) in Case 4, respectively. These finding suggest that co existence of extra-axial cyst such as Case 3 and 4 is incidental and that arachnoid cyst is less epileptogenic. However, intra-axial cyst such as Case 1 and 2 is epileptogenic and complicated physiological mechanism such as kindling phenomenon or secondary epileptogenesis may effect on the hippocampus. Comprehensive presurgical evaluation including electrocorticography is needed in the surgical treatment of TLE with cystic lesion. PMID- 12884804 TI - [A patient with Guillain-Barre syndrome and recurrent episodes of ST elevation and left ventricular hypokinesis in the anterior wall]. AB - A-65-year old woman who developed total ophthalmoplegia, areflexia in all her limbs and ataxia after upper respiratory tract infection was admitted to our hospital on the second day of illness. On day 3, she developed severe tetraparesis and respiratory failure which required mechanical ventilation, and Guillain-Barre syndrome (GBS) was diagnosed. On day 4, bilateral ptosis, facial diplegia, and neck muscle weakness appeared, and her all limbs were flaccid and immobile. An electrophysiological study suggested axonal damage. Marked blood pressure fluctuation also appeared on day 4. On day 5, an electrocardiogram showed a ST-segment elevation in leads V1 through V4 and the echocardiography showed anterior hypokinesia of the left ventricle. Her serum creatine kinase was normal. Left ventricular dysfunction and ST-segment elevation were normalized within hours, but a transient ST-segment elevation re-occurred on day 6. An electrocardiogram on day 13, showed inverted T waves in diffuse leads, which inversion continued. ST-segment elevation and hypokinesia in this patient were restricted to the left anterior descending branch, therefore, coronary spasm of that branch was considered the possible mechanism. In contrast, inverted T wave was due to either catecholamine cardiotoxicity or diffuse cardiac ischemia. Abnormalities of electrocardiogram were presumably due to cardiovascular autonomic dysfunction of GBS. PMID- 12884805 TI - [A case of neuro-Behcet's disease with repeating isolated thoracic spinal cord lesion]. AB - We reported a 45-year-old man who had repeated isolated thoracic spinal cord lesion on MRI in the clinical course of seven years. He had transient bilateral plantar numbness and urinary retention on December, 1994. Then, spastic paraplegia, total anesthesia of feet, and severe sphincter disturbance struck him on May, 1995. He was diagnosed as incomplete Behcet's disease and neuro-Behcet's disease on June, 1995 because of recurrent oral aphta, genital ulceration, and foliculitis. T2-weighted magnetic resonance imaging showed high intensity enhanced with Gd-DTPA in thoracic spinal cord from Th 5 to 8 level. Any other abnormal lesion on brain or spinal MRI was not observed. He was treated with corticosteroids and recovered incompletely. Another two big attacks occurred to him. No new lesion but thoracic spinal cord lesion was observed. We conclude that this case is the first reported example of neuro-Behcet's disease with repeating isolated thoracic spinal cord lesion. PMID- 12884806 TI - [A case of hypokalemic periodic paralysis: utility of exercise test for the assessment of therapeutic efficacy]. AB - We report a 14-year-old male with hypokalemic periodic paralysis. He noticed periodic paralysis at the age of 11. Any complication did not accompany the symptom. At the age of 12, hypokalemia was found during an episode of paralysis, and he was diagnosed as hypokalemic periodic paralysis. The frequency of paralytic attack increased around April 2000. Although long-acting oral potassium (32 mEq/day) was administered, it did not give favorable effect. Therapeutic spironolactone trial also failed. After the reconfirmation of the diagnosis of periodic paralysis by an exercise test, oral acetazolamide (750 mg/day) was started. In subsequent exercise test, the increment of the CMAP amplitude of abductor digiti minimi during exercise became smaller and the decrement of CMAP amplitude after exercise disappeared thereafter, which was assumed to be related with clinical improvement. The noninvasive exercise test is useful not only to diagnose periodic paralysis but also to evaluate therapeutic efficacy. PMID- 12884807 TI - [Amnesia, confabulation and nonaphasic misnaming after left thalamic infarct]. AB - A 72-year-old right handed woman developed amnesia, confabulation and abnormal (bizarre) verbal response after the left thalamic infarction. Clinical features including disorientation, euphoria and various kinds of paraphasia coincided in nonaphasic misnaming. MR images showed that lesions involved the genu of the internal capsule, the anteroventral nucleus, the lateroventral nucleus, intralaminar nuclei, the mamillothalamic tract and the region around the ventral thalamus. 99mTc-HMPAO-SPECT revealed decreased uptake in the left frontal lobe, probably due to the disconnection from the thalamus. These findings suggest that the dysfunction of the thalamus (mainly ventrolateral) and the left frontal lobe caused the disturbance of the self-monitoring in the language use, which generated confabulation and nonaphasic misnaming in our case. PMID- 12884808 TI - [Contrast enhanced fast fluid-attenuated inversion-recovery MR imaging for diagnosing cerebral venous angioma: report of two cases]. AB - It has been reported that contrast-enhanced fluid-attenuated inversion-recovery (FLAIR) sequences were useful for detecting superficial abnormalities, such as meningeal disease, because they do not demonstrate contrast enhancement of cortical vessels with slow flow as do T1-weighted images. We reported the usefulness of contrast-enhanced FLAIR images to differentiate cerebral venous angioma from tumor in two patients. Case 1 was a 71-year-old man developed cortical hemorrhage. Post contrast-enhanced T1-weighted images showed an enhanced lesion around the hematoma, whereas contrast-enhanced FLAIR images showed no enhancement of the lesion, thus he was diagnosed as cortical hemorrhage from cerebral venous angioma. Case 2 was a 72-year-old woman, who was examined MR images because of the jugular foramen neurinoma. There was a T2-high-intensity lesion in the right frontal lobe, and post contrast-enhanced T1-weighted images showed an enhanced lesion in and around the T2-high-intensity lesion. Post contrast FLAIR images showed no enhancement, and she was diagnosed as cerebral venous angioma. Contrast-enhanced fast FLAIR sequences was useful in differentiation between venous angiomas and tumors. Identification of these lesions was due to the flow-void phenomenon in vessels with slow-flowing blood such as venous angioma, which could not be differentiated from tumors on T1 weighted images. PMID- 12884809 TI - [The usefulness of Gadolinium-enhanced MR angiography for large cerebral aneurysm]. PMID- 12884810 TI - [Meningeal carcinomatosis]. PMID- 12884811 TI - [An autopsy case of amyotrophic lateral sclerosis with 22-years duration]. PMID- 12884812 TI - [Pathophysiological aspects of neutrophils--dual action of neutrophils]. PMID- 12884813 TI - [Cellular immunotherapy of cancer]. PMID- 12884814 TI - [Single-agent thalidomide for advanced and refractory multiple myeloma]. AB - Thalidomide as a single agent (200-400 mg/day) was administered in fourteen cases of refractory myeloma, from March 2001 till February 2002. The median age was 71 years (range 58 to 85 years), and the efficacy of thalidomide was observed in cases receiving treatment for at least three consecutive months. Response was evaluated in February 2002, according to the criteria for assessment of response described by Kakimoto et al. At the time of evaluation, two cases were in the PR2 state, one in PR3, two were stable, and three were PD. Evaluation of the response was not possible in six cases in whom treatment had to be discontinued due to intolerable side effects. The response to thalidomide was variable, with some cases responding well even to a low dose (200 mg/day) while a few others showed an early relapse due to the refractory nature of the disease in its response to the drug. The efficacy of treatment seemed to be correlated with the maturation pattern of myeloma cells. Side effects included neurological complications like somnolence, physiological symptoms such as constipation and so on, etc but all were relieved with symptomatic treatment. The drug was well tolerated in geriatric patients. Neutropenia was a dose limiting factor with half of the cases (7/14) presenting with severe neutropenia (grade 3-4), but a response was observed in all of them on administration of G-CSF. Thromboembolism occurred in two cases, the cause of which is not clear. These results suggest that thalidomide is a well tolerated drug and can be considered as a mainstay in the therapy of refractory myeloma. PMID- 12884815 TI - [A cold agglutinin disease, difficult to distinguish from paroxysmal cold hemoglobinuria]. AB - A 58-year-old male started passing dark brown colored urine in January 2002. An annual medical examination in the same month revealed a mild anemia and an increased serum LDH level. Because of the instability of the data of his peripheral red cell count and hemoglobin concentration as measuring with a blood cell auto-analyzer, and also the hemolytic findings in a test tube at room temperature (25 degrees C), he was referred to our hospital. Laboratory data revealed Hb 11.2 g/dl, reticulocytes 73.1% (233,000/microliter), indirect bilirubin 2.8 mg/dl, LDH 757 U/l, and hemosiderinuria, suggesting some intravascular hemolysis. The cold agglutinin titer was > 1,024, direct and indirect Coombs tests were both positive, and the Donath-Landsteiner antibody (D L antibody) was initially assessed as false positive because of a high titer of cold agglutinin. He was finally diagnosed as having a cold agglutinin disease (CAD) with anti-I autoantibody. Serologic tests for syphilis and mycoplasma, and various virus tests were all negatives. After avoiding cold exposure, his symptoms, including hemoglobinuria, disappeared. However, during 9 months follow up, he still showed a high titer of cold agglutinin. This case suggests that CAD should be considered when peripheral blood cell count data are unstable as assessed by a blood cell auto-analyzer. PMID- 12884816 TI - [Fatal septic shock and rhabdomyolysis following transfusion of platelet concentrates contaminated with Streptococcus pneumoniae]. AB - We report on a 58-year-old male diagnosed as having primary myelofibrosis with thrombocytopenia, who died of fatal septic shock and rhabdomyolysis after platelet concentrates (PCs) transfusion. The initial diagnosis of primary myelofibrosis was established by splenomegaly, leukoerythroblastosis and bone marrow fibrosis. PCs were transfused because of thrombocytopenia with marked bleeding tendency. Soon after the PCs transfusion in March 2000, he had attacks of chest pain, back pain and myalgia, then went into shock and died of unknown causes. PCs were suspected as being the cause of death, because Streptococcus pneumoniae was found in the culture of PCs in the WBC-reduction in-line filter and fresh frozen plasma from the same donor preserved in the Japan Red Cross Center. Rhabdomyolysis, neutrophil infiltration and phagocytosed bacteria were found from the autopsy materials, which were identified by DNA analysis as the same species found in the PCs. PCs are kept at room temperature because platelet function is lost in the cold. When PCs are contaminated with bacteria, marked multiplication induces fatal bacteremia. This is a rare report in Japan of fatal septic shock caused by PCs with bacterial contamination. We must pay strict attention to bacterial contamination in blood components. PMID- 12884817 TI - [Successful induction of complete cytogenetic response with high-dose imatinib mesylate and subsequent allogeneic stem cell transplantation for CML blastic crisis]. AB - A 29-year-old man was referred to our hospital with leukocytosis on March 7th, 2002. The white blood cell count was 132.9 x 10(3)/microliter with 42.0% blast cells. We diagnosed Philadelphia chromosome-positive chronic myelogenous leukemia (CML) in a blast crisis and started imatinib mesylate therapy at a dose of 800 mg/day on March 9th, 2002. The patient's peripheral blood blasts had disappeared by March 22nd, 2002, and the percentage of blasts in the bone marrow was 0.6% on May 2nd, 2002. The patient achieved a complete cytogenetic response on May 13th, 2002, and underwent allogeneic peripheral blood stem cell transplantation from his HLA-identical sibling donor on May 30th, 2002. Although adverse reactions such as grade 3/4 of hematological events (leukopenia, anemia, thrombocytopenia and neutropenia) and grade 1/2 of non-hematological events (hyperbilirubinemia, dermatitis and edema) were observed, these adverse reactions were clinically managed. This case suggested the usefulness of imatinib mesylate in the management of the CML-associated blast crisis. PMID- 12884818 TI - [Hemolytic uremic syndrome-like episode following nonmyeloablative hematopoietic stem cell transplantation in a patient with chronic myeloid leukemia]. AB - We report on a 61-year-old woman with chronic myeloid leukemia (CML) who developed a hemolytic uremic syndrome (HUS)-like episode following nonmyeloablative allogeneic hematopoietic stem cell transplantation. Macrohematuria, hypertension, hemolytic anemia with red cell fragmentation, thrombocytopenia, and progressive renal insufficiency were observed after thawed peripheral blood stem cell (PBSC) infusion. Although transient systemic hemolysis is known to occur during dimethylsulfoxide (DMSO)-cryopreserved stem cell infusion, HUS caused by DMSO has not been described in the literature. We speculate that one of the triggers of the HUS-like episode could have been renal microangiopathy caused by the long-term administration of interferon-alpha before the stem cell transplantation. PMID- 12884819 TI - [Myelodysplastic syndrome accompanied by Evans syndrome]. AB - Various cases of myelodysplastic syndrome (MDS) with diverse immunological disorders have been reported by many investigators. In this case report, we present a 70-year-old woman who had been diagnosed as having MDS and liver cirrhosis (LC) type C for 20 months, and who finally developed autoimmune phenomena against autologous blood cells. She was admitted to our hospital in order to evaluate her advanced anemia, thrombocytopenia and fatigue. The laboratory data at admission were as follows: hemoglobin 5.3 g/dl, red blood cell count 109 x 10(4)/microliter, white blood cell count 1,760/microliter, platelet count 4.3 x 10(4)/microliter and reticulocyte count 1.3%. The direct Coombs test was positive. In addition, anti-platelet antibody was positive, using the MPHA method. With these results, Evans syndrome secondary to MDS or LC was diagnosed. The mechanisms for the development of immunological disorders in patients with MDS or LC have not been fully elucidated. To our knowledge, this patient is one of the rare cases with MDS and Evans syndrome reported in the literature. During the generation process of autoantibodies, the role of HCV antigen in the pathogenesis of Evans syndrome was of interest in this patient. PMID- 12884820 TI - [Successful treatment of acute promyelocytic leukemia in a pregnant patient with all-trans retinoic acid and chemotherapy resulting in a safe delivery]. AB - A 32-year-old woman at 21 gestational weeks was admitted because of leukocytosis with DIC. She was diagnosed as having acute promyelocytic leukemia and treated with all-trans retinoic acid (70 mg/body) in combination with daunorubicin and cytosine arabinoside. She achieved complete remission, and continuously received a second treatment with daunorubicin and cytosine arabinoside. Cesarean section was performed, and a live male infant was born in the 30th week of pregnancy. The mother and baby have progressed excellently to date. In a such case, the choice of treatment and time of birth should be considered depending on the individual situation. PMID- 12884821 TI - [High dose ara-C therapy induced bradycardia in an acute myeloid leukemia patient with inv (16)(p13q22)]. AB - A 20-year-old Japanese woman was diagnosed as having acute myeloid leukemia with inv(16)(p13 q22). She achieved complete remission (CR) after treatment with a standard dose of cytarabine(ara-C) and idarubicin. She received high dose ara-C and etoposide for the 2nd consolidation chemotherapy. On the 5th fraction of high dose ara-C, her heart rate dropped to 52/minute and returned to 72/minute after the cessation of ara-C. After achieving informed consent, we gave her another course of high dose ara-C, which once again resulted in a decreased heart rate. This suggests that the administration of high dose ara-C could cause bradycardia. PMID- 12884822 TI - [Current status of reduced-intensity stem cell transplantation; survey in the Kinki area]. AB - The status of reduced-intensity stem cell transplantation (RIST) in the Kinki area was investigated by questionnaires. We sent out questionnaires to 72 institutions and received 51 replies (72.2%). RIST was performed in 16 institutions (31.4%). A total of 91 cases were included of whom approximately 70% were non-remission or advanced hematological malignancies. Various conditioning regimens were used for RIST, most often consisting of fludarabine, busulfan and antithymocte globulin. Acute GVHD developed in 57% of all patients. Relapse or disease progression developed in 42.7% of evaluable patients. The overall survival rate was 54.6%, and 80% of low risk patients were alive at the time of analysis. PMID- 12884823 TI - [A quantitative evaluation of spinocerebellar degeneration by an acoustic analysis--the effect of taltirelin hydrate on patients with Machado-Joseph disease]. AB - A clinical evaluation of 10 patients with Machado-Joseph disease (MJD) was performed by using an acoustic analysis soft (SoundScope, GW Instruments Inc) before and on 4 weeks of treatment of taltirelin hydrate (TH). A rapid repetitive monosyllable/ka/was recorded as the samples were analyzed in a computer. The repetition period (ms) and the maximum intensity (volt) was measured from the sound spectrogram and the amplitude envelope for each speech wave. Three acoustic parameters, which were mean of period (ms) (m-P), coefficient of variation of period (%) (CV-P), and coefficient of variation of amplitude (%) (CV-A), were calculated according to Ishida's acoustic analysis method. The average age in this study group was 52.4 +/- 11.4 years (32-73 years), the mean scores of International Cooperative Ataxia Rating Scale (ICARS) was 35.0 +/- 18.6. Significant correlation was shown between CV-A and ICARS scores before therapy (p < 0.05). CV-P was decreased significantly (p < 0.05) from 10.9 +/- 0.12% to 9.2 +/- 0.26% after TH, and its degree of decrement correlated with ICARS scores (p < 0.05), which means a greater effect of TH is expected in patients with milder symptoms. ICARS scores did not change with a statistical significance. TH was shown to be effective on the ataxic speech of patients with MJD by quantitative methods. The acoustic analysis used in this study is a sensitive and objective evaluation method for following up the clinical severity of MJD and judging effectiveness of a drug. PMID- 12884824 TI - [Clinical features of patients associated with anti-GM1b antibody]. AB - To clarify the clinical features of patients with anti-GM1b antibody, we retrospectively investigated which conditions were associated with anti-GM1b antibody in a large number of patients. Two hundred out of 1,713 patients had anti-GM1b antibody. One hundred and sixty-six (83%) were diagnosed with Guillain Barre syndrome (GBS) or atypical GBS with preserved deep tendon reflexes. Our study suggests that the anti-GM1b antibody testing is useful for supporting the diagnosis of GBS. In addition, anti-GM1b antibody was detected in 18 patients with Fisher syndrome and the related conditions associated with anti-GQ1b IgG antibody, and 3 patients with ataxic GBS associated with anti-GD1b IgG antibody. PMID- 12884825 TI - [The analysis of cytokines in cerebrospinal fluid (CSF) in two cases of non herpetic acute limbic encephalitis (NHALE)]. AB - We report two cases of non-herpetic acute limbic encephalitis (NHALE) which showed elevation of interleukin (IL)-6 in the cerebrospinal fluid (CSF). [Case 1] The patient was a 25-year-old woman who was admitted to another hospital because of fever and severe headache, following common cold. After the admission, she developed severe disturbance of consciousness and suffered from generalized convulsions, and was then transferred to our hospital. The CSF examination revealed neither pleocytosis nor elevation of total protein. Her consciousness improved by intravenous administration of high-dose methylprednisolone, but mild retrograde amnesia and symptomatic epilepsy remained as sequelae. [Case 2] The patient was a 58-year-old man who was admitted to our hospital because of fever, severe headache, and mild disturbance of consciousness, following common cold. After the admission, he exhibited marked psychiatric symptoms and severe amnestic syndrome. The CSF examination revealed mild lymphocytic pleocytosis and mild elevation of total protein. His clinical symptoms improved markedly by intravenous administration of high-dose methylprednisolone, but mild retrograde amnesia and personality changes remained. Cranial MRI showed reversible high signal intensity lesions in bilateral hippocampi and amygdaloid bodies on diffusion weighted images (DWI) in both cases. No laboratory findings suggesting herpes simplex virus infection or malignancy were detected in either case. In the CSF analysis of cytokines including IL-1 beta, IL-2, IL-6, IL-10, tumor necrosis factor alpha, and interferony gamma, only IL-6 was elevated in both cases. We recognized four clinical features in both cases as follows: 1. the episode of preceding infection such as common cold, 2. appearance of reversible high signal intensity lesions in bilateral hippocampi and amygdaloid bodies on DWI, 3. elevation of only IL-6 in CSF, and 4. marked neurological improvement by intravenous administration of high-dose methylprednisolone. We speculate that the immune reaction of the host might play some significant roles in the pathogenesis of NHALE, based on these four clinical features. PMID- 12884826 TI - [A case of Crow-Fukase syndrome with extramedullary plasmacytoma: marked clinical deterioration following a biopsy to plasmacytoma]. AB - A 66-year-old man developed paresthesia of the distal parts of the bilateral lower limbs a week after his upper respiratory infection, followed by the weakness with the legs and paresthesia with the lip area, tongue and finger tips. Those symptoms gradually became worse to the point that he was unable to walk 10 days later. Although skin pigmentation, edema, and lymph node swelling were not found, we made a diagnosis of Crow-Fukase syndrome (CFS) because of clinical features of polyneuropathy, IgG-lambda type M proteinemia, endocrinological abnormality, elevated plasma level of vascular endothelial growth factor (VEGF) and extramedullary plasmacytoma in his abdomen. Following intravenous immunoglobulin therapy (IVIg), he showed marked improvement. However, his neurologic symptoms deteriorated acutely just after open biopsy together with the elevation of VEGF level, and a few days later he was in the state of flaccid quadriparesis. We tried IVIg therapy again and his neurologic symptoms were markedly improved. We speculated that an elevated VEGF, released from plasma cells induced by the bioprocedure, might have caused an increase in microvascular permeability and affected the blood-nerve-barrier, thereby his neurologic symptoms deteriorated. It is thought that this case may support the hypothesis that a significant role is played by VEGF in the pathomechanism of the development of CFS. Additionally we experienced that IVIg was very effective to the neurologic symptoms, and we think that IVIg will be able to be one of the future therapy of the CFS. To our knowledge, there has been no report of CFS which manifested acute deterioration of his neurologic symptoms just after open biopsy with acute onset with Guillain-Barre syndrome like symptoms. PMID- 12884827 TI - [A Japanese case of Streptococcus suis meningitis associated with lumbar epidural abscess]. AB - A 47-year-old man working at a meat processing factory was admitted to our hospital after suffering high fever and severe headache for a 2-day period. He was alert and showed no neurological symptoms apart from neck stiffness. The CSF was turbid, with 701/microliter of polymorphnuclear leukocytes, a protein concentration of 218 mg/ml, and a sugar concentration of 33 mg/dl. He was diagnosed as suffering from Streptococcus suis (S. suis) serotype 2 meningitis based on CSF culture. Severe headache and high fever subsided after the treatment with intravenous antibiotics and immunoglobin. He developed, however, severe lumbargo on the third day, which was proven to be caused by the lumbar epidural abscess. Also, on the 8th day he developed hearing loss. These symptoms were treated and ameliorated with the administration of dexamethasone. On the 73rd day, the patient was discharged in good clinical condition, although he still suffered from a decrease in hearing perception. S. suis, found as a commensal in the nasopharynx of swine, is responsible for meningitis in the people who are in close contact with swine or unprocessed pork. S. suis meningitis seems to be the result of a preventable occupational infection. Although most of the patients come from other countries in Asia, our patient is the first documented case occurring in Japan. Those who have an occupational exposure to swine and pork are strongly recommended to use protective gloves. Prevention through public health surveillance is important. PMID- 12884828 TI - [MR spectroscopy findings of a case of intravascular malignant lymphoma: usefulness for differential diagnosis]. AB - We report a 49-year-old previously healthy woman with acute onset of decrease in attention, dysarthria and ataxia, accompanied by drowsiness. On admission, there were cloudness of consciousness, hallucination and left hemiparesis. Cerebrospinal fluid study revealed a cell count of 1/mm3, and the cytology was class I with a slight increase in protein. MRI of the brain performed on admission showed multiple gadolinium-enhanced lesions with a T2 weighted high intensity area in the cerebral white matter. At first the patient was diagnosed as acute disseminated encephalomyelitis (ADEM), and treated with methylprednisolon pulse therapy. Soon after, she showed transient clinical improvement, but her condition soon worsened. MR spectroscopy revealed elevated choline peak, decreased NAA peak and lactate peak, which indicated a neoplastic lesion. The brain biopsy disclosed diffuse intravascular lymphoma (IVL). MRS was useful in the differential diagnosis of IVL from ADEM. PMID- 12884829 TI - [Regional cerebral blood flow measured by three-dimensional stereotactic surface projections (3D-SSP) of 123I-IMP SPECT in Parkinson disease patients with cognitive impairment]. AB - We investigated the regional cerebral blood flow (rCBF) in 8 patients with Parkinson disease (PD) with cognitive impairment (age; 64-82 years, Mini-Mental State Examination score = MMSE score; 22-6 points, Yahr stage; III-V), with the standard transaxial images and the Z-score images using the three-dimensional stereotactic surface projections (3D-SSP) of 123I-IMP SPECT. A contrast database was created by averaging extracted database sets of the contrast group (numbers; 14 cases, age; 64-82 years, MMSE score; > or = 29 points). The regions of the perfusion reduction shown on the standard transaxial images were similarly demonstrated on the Z-score images in 6 of the 8 patients, and only the Z-score images demonstrated definite regions of perfusion reduction in remaining 2 patients. Both the standard transaxial and Z-score images demonstrated the perfusion reduction in the temporo-parietal regions in all of the patients, and the Z-score images but not the standard transaxial ones detected the reduction in the posterior cingulate gyrus and precuneus in 3 patients. 3D-SSP images of 123I IMP SPECT are thus more sensitive in detecting rCBF of the medial aspect of the parietal cortex than the standard transaxial images, and can be used as a diagnostic tool to objectively evaluate the cognitive function of PD patients. PMID- 12884830 TI - [Two sisters with dysferlinopathy manifesting different clinical phenotypes]. AB - We report two sisters with dysferlinopathy who manifested different clinical phenotypes. A 22-year-old female (patient 1) noticed of difficulty in running at the age of 13 years, and since then weakness of the lower extremities has progressed slowly. She had typical features of Miyoshi myopathy (MM); i.e., young adult onset, dominant involvement of calf muscles and markedly elevated serum creatine kinase (CK). Her 19-year-old sister (patient 2) first noticed of weakness in lower extremities at age 12 years. On neurological examination, she had proximally dominant muscle weakness, consistent with limb girdle muscular dystrophy type 2B (LGMD2B); serum CK level was also markedly elevated. On MRI study of muscle, patient 1 showed fatty degeneration of calf muscles, whereas patient 2 showed no abnormality in quadriceps and calf muscles. Immunohistochemistry of the muscle biopsy specimens using anti-dysferlin antibody showed deficiency of this protein in sarcolemma. There have been only a few reports of sibling cases of dysferlinopathy whose clinical phenotypes are different. These sibling cases may have important suggestion on the mechanism(s) of phenotypic variation of dysferlinopathy. PMID- 12884831 TI - [A case report of Parkinson's disease presenting with recurrent dyspneic attacks due to focal laryngeal dystonia]. AB - A 69-year-old woman suffering from Parkinson's disease for 22 years was admitted because of frequent occurrence of paroxysmal dyspnea for 3 months. Her dyspneic attacks consisting of inspiratory stridor and cyanosis occurred mainly during the wearing-off time and continued for less than 30 min. During nonictal period her respiration and phonation were normal and endoscopic investigation of the vocal cord and upper respiratory tract revealed no abnormality. Based on these findings, she was diagnosed to have focal laryngeal dystonia. The 24-hr monitoring with pulseoxymeter recorded frequent occurrence of paroxysmal asymptomatic hopoxemia during both daytime and sleep. With the treatment of tracheostomy and the reduction and alteration of anti-Parkinsonian drugs, dyspneic attacks disappeared gradually. We also confirmed the complete disappearance of paroxysmal asymptomatic hopoxemia with the 24-hr monitoring by pulseoxymeter, which is considered to be a useful method for early detection of asymptomatic focal laryngeal dystonia. PMID- 12884832 TI - [Bilateral medial medullary infarction presented with monoplegia of the lower limb, followed by paraplegia and finally by tetraplegia]. AB - A 69-year-old Japanese man suddenly developed monoplegia of left lower extremity, followed by paraplegia and finally by tetraplegia. MRI revealed an infarction in bilateral medial medulla extending from the cervicomedullary junction up to the upper limit of the medulla. Both hypoglossal nerve palsy and sensory disturbance were absent. At the pyramidal decussation, fibers to the lower extremities cross caudal to the fibers going to the upper extremities, therefore right below the decussation, fibers to the lower extremities run medial side of the fibers to the upper extremities, but later the former run lateral side of the latter. In this patient, the authors considered that the lesion initially damaged the pyramidal decussation at a slightly lower level, involving the tract to left lower extremity, and then extended to right lower extremity, to the left upper extremity, finally to the right upper extremity. Bilateral medial medullary infarction must be considered in the clinical course seen as in this patient. PMID- 12884833 TI - Family history and coronary artery disease. PMID- 12884834 TI - Back from the Peace Corps--with skin lesions from South America. PMID- 12884835 TI - [Human hepatitis B virus infection of tree shrews and Macaca assamensis in vivo]. AB - OBJECTIVE: To examine sensitivity of the tree shrews and Macaca assamensis to human hepatitis B virus (HHBV) by serologic methods. METHODS: Totally 233 tree shrews and 28 Macaca assamensis were inoculated with human sera containing HBV. After inoculation, the sera were collected weekly from them and HBV markers were detected with HBV ditecting ELISA kits. RESULTS: Ninety percent of the tree shrews developed acute infection, among them, 44.4 % persisted for over one year, 33.3% of them developed chronic infection persisted for 2 years and one month; the persistence of HBV in Macaca assamensis was much shorter. CONCLUSION: These data clearly indicated that tree shrew may be used as an animal model for study of chronic HBV infection, whereas, Macaca assamensis, showed only a transient sensitivity to HHBV. PMID- 12884836 TI - The role of colour in implicit and explicit memory performance. AB - We present two experiments that examine the effects of colour transformation between study and test (from black and white to colour and vice versa, of from incorrectly coloured to correctly coloured and vice versa) on implicit and explicit measures of memory for diagnostically coloured natural objects (e.g., yellow banana). For naming and coloured-object decision (i.e., deciding whether an object is correctly coloured), there were shorter response times to correctly coloured-objects than to black-and-white and incorrectly coloured-objects. Repetition priming was equivalent for the different stimulus types. Colour transformation did not influence priming of picture naming, but for coloured object decision priming was evident only for objects remaining the same from study to test. This was the case for both naming and coloured-object decision as study tasks. When participants were asked to consciously recognize objects that they had named or made coloured-object decisions to previously, whilst ignoring their colour, colour transformation reduced recognition efficiency. We discuss these results in terms of the flexibility of object representations that mediate priming and recognition. PMID- 12884837 TI - Preparing those next at bat. PMID- 12884839 TI - Acetylcysteine for prevention of acute deterioration of renal function following elective coronary angiography and intervention: a randomized controlled trial. PMID- 12884838 TI - The human vomeronasal organ. V. An interpretation of its discovery by Ruysch, Jacobson, or Kolliker, with an English translation of Kolliker (1877). AB - The vomeronasal organs (VNOs) of mammals are highly variable epithelial structures found bilaterally in the mucosa of the nasal septum. Whereas the discovery of the human VNO is traditionally ascribed to Frederick Ruysch (1703, 1724), the organ is named after Ludwig Levin Jacobson (1811, 1813) who described it in nonhuman mammals. We recently have pointed out controversies surrounding the incidence and structure of the enigmatic human VNO, and herein, we provide a historical analysis of its discovery. We present evidence that the honor of discovering the human VNO truly belongs to Kolliker (1877), and not to Ruysch. Ruysch illustrated the lateral view of a 2-year-old infant's nasal septum, and it is unclear whether the right nasal passage, the tubular VNO or its opening, or an unrelated duct is being indicated. Jacobson reported the VNO to be missing in humans. Its discovery in the human embryo can be related in part to later authors, such as Dursy (1869). Our reappraisal of the literature confirms that Kolliker was actually the first among these 18th-689th century investigators to provide evidence of the human VNO as a histologically identifiable structure in the fetus and the adult. PMID- 12884840 TI - PET on display: notes from the 50th SNM annual meeting. PMID- 12884841 TI - PET and PET/CT: market outlook. PMID- 12884842 TI - PET_Mail: a low-key, high-tech PET community. PMID- 12884843 TI - SNM highlights as history: 1983. PMID- 12884844 TI - The height-based formula for prediction of left-sided double-lumen tracheal tube depth. PMID- 12884845 TI - The timing for the use of heparinase and tromboelastography to prevent excessive bleeding coronary artery bypass graft surgery. PMID- 12884846 TI - Should first-time traumatic shoulder dislocations undergo an acute stabilization procedure? PMID- 12884847 TI - Lifting the profile of nutrition in Australia. PMID- 12884848 TI - Military health care and telemedicine. PMID- 12884849 TI - Endoscopic mucosal resection (EMR) for the management of poorly differentiated adenocarcinoma of the stomach: a patient who had recurrence and died 4 years after EMR. AB - Recent studies have explored the possibility of using endoscopic mucosal resection (EMR) to treat lesions with a relative indication for endoscopic therapy. We used EMR to manage poorly differentiated adenocarcinoma, a relative indication for endoscopic treatment, in a patient who requested such treatment and refused surgical intervention. We describe our experience with this patient, who died of tumor recurrence 4 years after the EMR. This case describes the problems of using EMR for the treatment of poorly differentiated adenocarcinoma. PMID- 12884850 TI - Weekly paclitaxel for a patient with advanced gastric cancer. AB - A 56-year-old woman diagnosed with gastric cancer was admitted to our hospital for operation on May 15, 2001. The operation was performed on May 23. The tumor formed a large mass from the antrum to the head of the pancreas, and cancer cells were detected in the ascitic fluid microscopically. During the operation, resection was impossible, and so 100 mg of cisplatin (CDDP) was infused into the abdominal cavity. After the operation, she experienced continuous nausea and there was a discharge of 1200-1600 ml of digestive fluid per day from her nasogastric tube. On July 17, a new regimen, of 4-week courses of chemotherapy, with weekly administrations of 65 mg/m2 of paclitaxel, along with premedication for 3 weeks, followed by 1 week of rest, was started. After the first of these 4 week courses, the discharge from her nasogastric tube decreased to 200-600 ml per day, and the tube was removed 78 days after insertion. Oral intake of food increased smoothly, and she was discharged on September 14. After another, short, hospitalization, she was discharged on October 20, and she has been coming to our outpatient clinic once a week. After paclitaxel was started, gastric fiberscopy and computed tomography (CT) scan showed reduction of the tumor. Of special note was the disappearance of a scitic fluid after two courses, rated as a "partial response" (Japanese classification). There was a decrease in hemoglobin, but neither leukocytopenia nor a decrease in platelets was found. Neuropathy was slight and no treatment was needed. Now, after 1 year, 11 courses of chemotherapy have been administered at the outpatient clinic. These results suggest weekly administration of paclitaxel to be a promising treatment for advanced gastric cancer with peritoneal dissemination. The therapeutic efficacy should be confirmed by further clinical trials. PMID- 12884851 TI - Almost one in three non-elderly Americans uninsured for all or part of 2001-2002. PMID- 12884852 TI - Health consequences of lack of health insurance. PMID- 12884853 TI - Community consequences of lack of health insurance. PMID- 12884854 TI - Radio daze: young people targeted by alcohol ads. PMID- 12884855 TI - Essential role for IKKgamma/NEMO in TCR-induced IL-2 expression in Jurkat T cells. AB - The control of IL-2 gene expression in T cells by multiple transcriptional factors has been extensively explored, however, the role of the NF-kappaB signaling pathway in TCR-dependent IL-2 production still remains unclear. In this study, we used a somatic cell genetics approach to address this question. Triggering TCR in mutant Jurkat T cells lacking IKKgamma/NEMO failed to induce IL 2 due to a selective loss in I-kappaB kinase activity, I-kappaBalpha degradation and NF-kappaB DNA-binding activity. The AP-1 and NF-AT binding activities in the IL-2 promoter were comparable between wild-type and mutant T cells. These defects in the mutant cell line were rescued by the reintroduction of exogenous IKKgamma. Taken together, our data demonstrate that IKKgamma plays an essential role in TCR induced signaling pathways leading to IL-2 expression. PMID- 12884856 TI - Granulysin in human serum as a marker of cell-mediated immunity. AB - Granulysin is a cytolytic granule protein of natural killer (NK) cells and cytotoxic T lymphocytes (CTL) with a broad range of antimicrobial and tumoricidal activities. Two molecular forms of granulysin, the 15-kDa precursor and 9-kDa mature form, are produced in these cells. In this study, we developed monoclonal antibodies against granulysin and found that the 15-kDa granulysin is spontaneously secreted by peripheral blood NK and T cells via a non-granule exocytotic pathway. When NK cells killed the target cells, the released granulysin levels in culture supernatants significantly increased through the granule exocytosis. The granulysin protein was found in the sera of healthy individuals at an average concentration of 3.7 +/- 3.2 ng/ml (age 0-99 years, n=244). The serum levels of granulysin were transiently highly elevated among patients with acute viral infections. In addition, the serum granulysin levels in patients with severe immunodeficiency treated bycell therapy fluctuated proportionately to the improvement of other immunological parameters. Our results suggest that granulysin is well associated with diverse activities of NK cells and CTL in physiological and pathological settings and could be a useful novel serum marker to evaluate the overall status of host cellular immunity. PMID- 12884857 TI - Imatinib (STI-571) heals a gastrocutaneous fistula resulting from a malignant gastric stromal tumor. AB - The management of inoperable giant malignant gastrointestinal stromal tumor (GIST) of the stomach sed to be a formidable task. We report our success with the use of STI-571 in treating a patient with huge GIST of the stomach complicated by gastrocutaneous fistula after an unsuccessful laparotomy. The patient was a 49 year-old man who presented to our center with a painful epigastric mass in December 2001. Endoscopy, biopsy, and magnetic resonance scan confirmed that it was a malignant stromal tumor arising from the gastric fundus. Laparotomy with an intention to resect the tumor was performed in view of the obstructing symptoms. However, massive bleeding was encountered during dissection of the tumor and gastrectomy was abandoned. The case was further complicated with the development of a gastrocutaneous fistula in the early postoperative period. The patient ws then managed with naso-duodenal tube feeding and enteral STI-571 was prescribed. The fistula healed up in 20 days and the mass became impalpable 1 month afterwards. Follow-up computed tonography (CT) scan 3 months later confirmed significant tumor reduction, and the patient has experienced no side effects from the treatment PMID- 12884858 TI - Xiphoidectomy. AB - Complex surgical procedures in the upper abdomen required ear exposure. To improve exposure xiphoidectomy is indicated; however, in the past there was an extensive time commitment and unpredictable bleeding. We describe a rapid and bloodless approach to xiphoidectomy, utilizing a self-retaining retractor, controlled osteotomy, and complete removal to the xiphisternal junction. The xiphoid bone is dissected with high-voltage electrosurgery, denatured, and resected completely. Bleeding points are easily controlled with electrocoagulation. This procedure allows optimal exposure to the most superior aspect of the abdominal cavity. PMID- 12884859 TI - Differential acquired immune responsiveness to bacterial lipoproteins in Lyme disease-resistant and -susceptible mouse strains. AB - We investigated the effect of Borrelia burgdorferi lipoproteins (outer surface protein A) and the synthetic lipohexapeptide tripalmitoyl-S-glyceryl-Cys-Ser 4(Lys) (Pam3-Cys) on isolated lymph node (LN) cells from Lyme disease-susceptible (C3H/HeJ) and -resistant (C57BL/6J) mice. Mice were either infected with B. burgdorferi for 1 week or left uninfected. Lipoprotein-stimulated LN cells from infected C3H/HeJ mice produced significantly higher levels of the inflammatory cytokines IL-6 and IFN-gamma than did cells from C57BL/6J mice. Cells from uninfected mice did not respond. No TNF-alpha or IL-1beta were produced by LN cells from infected mice of either strain in response to lipoprotein or B. burgdorferi spirochetes. Unlike with IL-6 or IFN-gamma, LN cells from either strain failed to produce IL-10 in response to lipoproteins. However, the LN cells were able to produce this cytokine in response to B. burgdorferi spirochetes or after incubation with phorbol-12-myristate-13-acetate/ionomycin, anti-CD3 antibody alone or anti-CD3 combined with anti-CD28 antibodies. Addition of exogenous IL-10 to lipopeptide-stimulated cultures significantly reduced IFN gamma and IL-6 production in a dose-dependent fashion. This inhibition was more effective with cells from disease-resistant C57BL/6J mice than with cells from disease-susceptible C3H/HeJ mice. The proclivity to disease of the C3H/HeJ mouse could be simultaneously based on the phenomena of enhanced inflammatory responsiveness to lipoproteins and diminished ability to respond to IL-10. An investigation of the determinants of these two phenomena could be used as a blueprint to elucidate the pathogenesis of Lyme disease in humans. PMID- 12884860 TI - Incomplete effector/memory differentiation of antigen-primed CD8+ T cells in gene gun DNA-vaccinated mice. AB - DNA vaccination is an efficient way to induce CD8+ T cell memory, but it is still unclear to what extent such memory responses afford protection in vivo. To study this, we induced CD8+ memory responses directed towards defined viral epitopes, using DNA vaccines encoding immunodominant MHC class I-restricted epitopes of lymphocytic choriomeningitis virus covalently linked to beta2-microglobulin. This vaccine construct primed for a stronger recall response than did a more conventional minigene construct. Despite this, vaccinated mice were only protected against systemic infection whereas protection against the consequences of peripheral challenge was limited. Phenotypic analysis revealed that DNA vaccine-primed CD8+ T cells in uninfected mice differed from virus-primed CD8+ T cells particularly regarding expression of very-late antigen (VLA)-4, an adhesion molecule important for targeting T cells to inflammatory sites. Thus, our DNA vaccine induces a long-lived memory CD8+ T cell population that provides efficient protection against high-dose systemic infection. However, viral replication in solid non-lymphoid organs is not curtailed sufficiently fast to prevent significant virus-induced inflammation. Our results suggest that this is due to qualitative limitations of the primed CD8+ T cells. PMID- 12884861 TI - Thymic epithelial cells provide WNT signals to developing thymocytes. AB - Interactions with thymic stromal cells are known to be critical for the development of T cells from progenitors entering the thymus, yet the molecular mechanisms of stromal cell function remain poorly understood. Accumulating evidence has highlighted the importance of beta-catenin-mediated activation of T cell factor (TCF)/lymphoid enhancer factor (LEF) transcription during thymocyte development. As regulation of this signaling pathway is controlled by binding of soluble Wnt proteins to cell surface Frizzled (Fz) receptors, we studied components of Wnt/Fz-mediated signaling in thecontext of stromal cell regulation of thymocyte development. We show that mRNA for a variety of Wnt family members, notably Wnt-4, Wnt-7a and 7b, and Wnt-10a and 10b, are expressed by thymic epithelium rather then by thymocytes, while thymocytes demonstrate a developmentally regulated pattern of Fz receptor expression. Collectively these findings suggest (1) a functional role for Wnt-producing thymic epithelium in determining TCF/LEF-mediated transcriptional regulation in Fz-bearing thymocytes, and (2) a role for defined Wnt-Fz interactions at successive stages of thymocyte maturation. In support of this we show that separation of thymocytes from Wnt producing epithelial cells and the thymic microenvironment, triggers beta-catenin phosphorylation and degradation in thymocytes. Thus, sustained exposure to Wnt in the context of an intact stromal microenvironment is necessary for stabilization of beta-catenin-mediated signaling in thymocytes. PMID- 12884862 TI - The efficacy of complement-mediated phagocytosis of Cryptococcus neoformans is dependent on the location of C3 in the polysaccharide capsule and involves both direct and indirect C3-mediated interactions. AB - Complement component 3 (C3) is the major opsonin for the pathogenic fungus Cryptococcus neoformans in the non-immune host. However, the efficiency of complement-mediated opsonization varies, depending on the strain, through mechanisms that are not understood. Analysis of complement-mediated phagocytosis for 12 strains grown in Sabouraud medium revealed that phagocytic indices were inversely correlated with capsule volume. In contrast, there was no correlation between phagocytic index and capsule volume for IgG1-opsonized cells. When capsule size was increased, the efficacy of complement-mediated phagocytosis decreased, whereas that of antibody-mediated phagocytosis increased. C3 localized inside the capsule and at the outer capsule edge for poorly phagocytozed and well phagocytozed strains, respectively. Blocking experiments revealed that complement mediated phagocytosis occurred through complement receptor 3 (CR3), without significant involvement of CR1 or CR4. Blocking experiments with antibodies to C3 did not completely abrogate yeast cell uptake, consistent with phagocytosis through glucuronoxylomannan-CR3 interactions. Our data explain how some large encapsulated cells avoid phagocytosis and suggest a novel strategy for immune evasion whereby a microbial capsule interferes with phagocytosis by modifying the location of C3 deposition. PMID- 12884863 TI - Pre-TCR signaling regulates IL-7 receptor alpha expression promoting thymocyte survival at the transition from the double-negative to double-positive stage. AB - The pre-T cell receptor (pre-TCR) and IL-7 receptor (IL-7R) are critical mediators of survival, proliferation and differentiation in immature thymocytes. Here we show that pre-TCR signaling directly maintains IL-7Ralpha expression as developing thymocytes undergo beta-selection. Inhibition of IL-7/IL-7R signaling in (CD44-CD25-) DN4 cells results in decreased generation of double-positive thymocytes due to increased death of rapidly proliferating beta-selected cells. Thus, we identify a mechanism by which pre-TCR signaling controls the selective survival of TCRbeta+ thymocytes, and define a further stage of T cell differentiation in which signaling from a TCR regulates the ability of that cell to respond to cytokine. PMID- 12884864 TI - Nucleosomes induce lymphocyte necrosis. AB - Nucleosomes are major autoantigens in systemic lupus erythematosus. These are found as circulating complexes in both lupus patients and lupus mice. The existence of a potential ubiquitous cell surface receptor specific for nucleosomes has been suggested. However, neither the nature nor the role of this receptor has been identified. Moreover, the consequence of receptor-nucleosome interaction on the target cell has not been studied in detail so far. We show here that purified nucleosomes induce cell death of normal and lupus lymphocytes ex vivo in a dose- and time-dependent manner whereas human dendritic cells were relatively resistant. Most importantly, nucleosome-induced cell death is primary necrosis. Moreover, necrosis could be abolished when nucleosomes were first treated with deoxyribonuclease I, proteinase K or with a specific monoclonal antibody. Finally, intravenous injections of purified nucleosomes result in a reduced spleen cell number compared to buffer-injected mice, indicating that circulating nucleosomes may behave similarly in vivo. Taken together, this is the first report indicating that nucleosomes are able to induce necrosis,which in turn could result in an amplification loop of the disease causing the inflammation observed in lupus patients. PMID- 12884865 TI - Suppressor activity of anergic T cells induced by IL-10-treated human dendritic cells: association with IL-2- and CTLA-4-dependent G1 arrest of the cell cycle regulated by p27Kip1. AB - We have previously shown that human IL-10-treated dendritic cells (DC) induce an antigen-specific anergy in CD4+ T lymphocytes. These anergic T cells are characterized by an inhibited proliferation, a reduced production of IL-2, and additionally display antigen-specific suppressor activity. In this study we investigated the mechanisms underlying the anergic state and regulatory function of these T cells. We did not observe enhanced rates of programmed cell death of anergic CD4+ suppressor T cells compared to T cells stimulated with mature DC. Cell cycle analysis by DNA staining and Western blot experiments revealed an arrest of anergic CD4+ T suppressor cells in the G1 phase. High levels of the IL 2-dependent cyclin-dependent kinase (cdk) inhibitor p27Kip1 were found in anergic CD4+ suppressor T cells resulting in an inhibited activation of retinoblastoma protein and an arrest of cell cycle progression in the G1 phase. Addition of IL 2, but not blocking of the CTLA-4 pathway restored the proliferation of the suppressor T cells. In contrast, both treatments induced a down-regulation of p27Kip1 and acomplete inhibition of the antigen-specific regulatory function as demonstrated by high proliferation and enhanced IFN-gamma production of co cultured T cells. Further experiments demonstrated that p27Kip-expressing regulatory CD4+CD25+ T cells did not contribute to induction of T cell anergy in this model. Our data show that regulatory function of anergic CD4+ suppressor T cells is associated with an arrest in the G1 phase of the cell cycle mediated by increased levels of the IL-2- and CTLA-4-dependent cdk inhibitor p27Kip1. PMID- 12884866 TI - Caspase-8 and caspase-10 activate NF-kappaB through RIP, NIK and IKKalpha kinases. AB - NF-kappaB regulates the expression of various genes involved in cell growth and differentiation, immune response and inhibition of apoptosis. Recently, some death effector domain (DED)-containing proteins, such as FADD and c-FLIP were reported to activate NF-kappaB. We previously reported that the prodomain-only isoforms of caspase-8 and -10 (PDCasp8/10), containing two DED motifs, could inhibit Fas-mediated apoptosis. Here, we demonstrate that these isoforms also activate NF-kappaB, implying this to be one of the mechanisms by which these polypeptides inhibit apoptosis. The GST pull-down assay revealed that, among upstream kinases that activate NF-kappaB, only NIK and RIP, but not RICK or IKKalpha/beta, could directly bind to PDCasp8/10. In addition, both modules ofDED in PDCasp8/10 were required for these interactions as well as NF-kappaB activation. Experiments using a kinase-dead mutant of IKKalpha and an RIP mutant lacking a kinase domain, both of which function as dominant-negative mutants for their wild-type counterparts, blocked PDCasp8/10-mediated NF-kappaB activation. Using small interfering RNA technology, we further demonstrate that the down regulation of IKKalpha but not IKKbeta significantly inhibits PDCasp8-mediated NF kappaB activation. Taken together, these results suggest that caspase-8 and -10 have roles in a non- or anti-apoptotic signaling pathway leading to NF-kappaB activation through RIP, NIK and IKKalpha. PMID- 12884867 TI - Type-1 polarized nature of mouse liver CD8alpha- and CD8alpha+ dendritic cells: tissue-dependent differences offset CD8alpha-related dendritic cell heterogeneity. AB - Interleukin-12 p70 (IL-12p70) is a major dendritic cell (DC)-produced cytokine known to support type-1 T helper (Th1) cells and inflammatory-type immunity. While the ability of DC to produce bioactive IL-12p70 depends on both the DC subtype and the microenvironmental conditions of DC development, the relative contribution of each of these factors remains unclear. Here, we report that in contrast to spleen CD8alpha+ and CD8alpha- DC that show strong differences in their respective IL-12p70-producing capacities, CD8alpha+ and CD8alpha- DC isolated from the liver, a non-lymphoid organ, both efficiently produce IL-12p70 in amounts comparable to spleen CD8alpha+ DC. The IL-12p70-producing capacity CD8alpha+ and CD8alpha- DC from either location is greatly increased following their overnight culture in the presence of granulocyte-macrophage colony stimulating factor. The elevated production of IL-12p70 by short-term cultured DC correlates with their enhanced expression of CD40 and other costimulatory molecules, and elevated T cell-stimulatory capacity. These data indicate that low IL-12-producing capacity is not an intrinsic property of the CDalpha8- DC subtype, and support the hypothesis that factors such as the site of DC development and maturation stage play a dominant role in defining DC function. PMID- 12884868 TI - H2-O influence on antigen presentation in H2-E-expressing mice. AB - MHC class II molecules sample peptides generated in the endosomal/lysosomal system for cell surface presentation to CD4+ T cells. Peptide loading requires the endosomal/lysosomal resident HLA-DM (DM; H2-DM, mouse), but in B cells, DM is tightly associated with HLA-DO (DO; H2-O, mouse). We have previously shown that H2-O differentially modulates the processing and presentation of different antigenic epitopes on H2-Ab molecules. Using H2-Ead-transgenic mice, we here show that presentation of different epitopes by H2-Ed/b molecules is similarly influenced by H2-O after membrane immunoglobulin-mediated uptake of antigen. In addition, B cells from H2-Ead-transgenic mice (which co-express H2-Ab and H2-Ed/b molecules) show an altered pattern of presentation of H2-Ab-restricted epitopes. In H2-Ead-transgenic, H2-O-deficient mice, further changes in the peptide repertoire were observed. Thus, H2-Ed/b expression influences the epitopes presented by H2-Ab, and this effect is further altered by expression of H2-O. PMID- 12884869 TI - Recovery from EAE is associated with decreased survival of encephalitogenic T cells in the CNS of B7-1/B7-2-deficient mice. AB - Adoptive transfer experiments using C57BL/6 mice lacking B7-1 and B7-2 as recipients of wt (wt) encephalitogenic T cells demonstrate a key role for B7 costimulation during the effector phase of experimental autoimmune encephalomyelitis (EAE). Following transfer of encephalitogenic T cells, B7-1/B7 2-deficient (-/-) recipients develop a transient and mild disease as compared to wt recipients. To understand the mechanism by which B7-1/B7-2 may influence the effector phase of EAE, we analyzed T cells, pro-inflammatory cytokines and chemokines within the CNS of wt and B7-1/B7-2-/- recipients at different times after adoptive transfer of activated myelin specific T cells. There was a marked decline in T cells and inflammatory mediators in the CNS of B7-1/B7-2-/- recipients by day 30 post transfer. B7-1/B7-2-/- mice developed more TUNEL+ apoptotic cells in the parenchyma and greater ratios of TUNEL+ cells/parenchymal foci than wt mice resulting in virtual disappearance of parenchymal foci. Therefore, without B7-1 and B7-2 costimulation in the target organ, there is increased T cell apoptosis and attenuation of inflammation. These results indicate that B7-1 and B7-2 provide critical costimulatory signals for sustaining survival of pathogenic T cells within the central nervous system parenchyma during the effector phase of EAE and suggest novel treatment approaches in the effector phase of autoimmune diseases. PMID- 12884870 TI - NKG2D engagement of colorectal cancer-specific T cells strengthens TCR-mediated antigen stimulation and elicits TCR independent anti-tumor activity. AB - The NKG2D receptor is expressed by human NK, gammadelta T and alpha/beta T lymphocytes and its engagement results in the stimulation of effector cells. We evaluated the role of NKG2D receptor in anti-colorectal cancer (CRC) immune response. The cell surface expression of stress-inducible NKG2D ligands MICA/B (MHC class I-related chain molecules A/B) and ULBP (UL16 binding protein) by a panel of CRC lines was evaluated by flow cytometry. MICA and ULBP2/3 were widely expressed by the analyzed lines, with a minority of them being also ULBP-1+, whereas MICB was undetectable. CD8+ and CD4+ HLA-restricted anti-tumor T cell clones of a CRC patient were used to evaluate whether NKG2D engagement could mediate tumor recognition. Three out of four CD8+ T cell clones recognized the autologous tumor with a marginal NKG2D engagement, a finding that was correlated with the weak expression of NKG2D ligands by the autologous tumor. On the contrary, NKG2D triggering of these CD8+ T cell clones induced recognition of allogeneic CRC lines showing high expression of MICA and ULBP. A costimulatory role of NKG2D was observed with one CD4+/NKG2D+ T cell clone when stimulated by tumors sharing the HLA class II alleles and expressing NKG2D ligands. Taken together these data indicate that the engagement of NKG2D, depending on the expression of its ligands by target cells, can influence the pattern of anti tumor reactivity by T lymphocytes. PMID- 12884871 TI - The ester-bonded palmitoyl side chains of Pam3CysSerLys4 lipopeptide account for its powerful adjuvanticity to HLA class I-restricted CD8+ T lymphocytes. AB - Molecularly defined adjuvants are urgently required to implement immunization protocols by which CD8+ T cells induction is envisaged. We show here that the synthetic lipopeptide Pam3CysSerLys4 (P3CSK4) strongly enhances the expansion of antigen-specific IFN-gamma+CD8+ cells in vitro. These effects critically depend on the presence of two ester-bonded palmitoylated side chains. In fact, T cell expansion is impaired in the presence of derivatives bearing two non palmitoylated fatty acid chains, while derivatives with only one amide-bonded palmitoylated residue are completely inactive and behave like the non-lipidated peptide backbone. P3CSK4 is not mitogenic for T lymphocytes and can modulate DC immune biological properties. Indeed, doses as low as 100 ng/ml increase CD86, CD83 and CD40 surface expression on DC, fail to induce CCR7, and trigger a defined pattern of soluble factors associated to immune effector functions. In particular, substantial amounts of TNF-alpha, IL-6, CCL2 and CXCL10, in the absence of IFN-alpha, IFN-gamma, IL-15, IL-12p70 and CX3CL1, can be measured. Accordingly, antigen-specific CD8+ T cells expanded in vitro express CCR2 and CXCR3 chemokine receptors. Altogether our data suggest that human DC are able to respond to chemically different synthetic lipopeptide analogs and that optimal adjuvanticity to CD8+ T cell induction is achieved by the palmitoylated structures. PMID- 12884872 TI - TNF-alpha induces the generation of Langerin/(CD207)+ immature Langerhans-type dendritic cells from both CD14-CD1a and CD14+CD1a- precursors derived from CD34+ cord blood cells. AB - CD34+ cell-derived hematopoietic precursors amplified with FLT3-ligand, thrombopoietin and stem cell factor became, after a 6-day induction with GM-CSF, IL-4 and TGF-beta1, HLA-DR+, CD1a+, CD83-, CD86-, CD80- cells. A fraction of them expressed Langerin, Lag, and E-cadherin, resembling epidermal Langerhans cells (LC). TNF-alpha added for the last 3 days only marginally induced CD83 expression, but strikingly increased the proportion of immature Langerin+CD83- LC. Langerin+CD83+ and Langerin+CD83- cells were functionally distinct, the former internalizing less efficiently Langerin than the latter. Both CD1a-CD14- and CD1a-CD14+ cells sorted from FLT3-ligand, thrombopoietin and stem cell factor cultures responded to TNF-alpha by an increase of Langerin+ cells. Thus, TNF alpha rescued LC precursors irrespective of their commitment to the monocytic lineage. When added to GM-CSF, IL-4 and TGF-beta1 containing-cultures, LPS or IL 1beta also induced significant numbers of Langerin+CD83- immature cells displaying a low allostimulatory activity, while CD40-ligand largely promoted highly allostimulatory Langerin-CD83+ cells. Altogether, these data show that in contrast to CD40-ligand, which induced LC maturation even in presence of TGF beta1, nonspecific proinflammatory factors such as TNF-alpha, IL-1 or LPS, essentially induced immature LC generation, and little cell activation in the presence of TGF-beta1. PMID- 12884873 TI - [Clinical problems in medical mycology: problem #3. Paracoccidioidomycosis]. PMID- 12884874 TI - Lymphadenopathy of sinus histiocytosis mimicking metastatic bladder cancer: a case report. AB - A case of invasive bladder cancer accompanied with enlarged pelvic lymph nodes, which were interpreted as metastasis, is reported. The lymphadenopathy eventually regressed spontaneously while the bladder cancer increased in size. Total cystectomy and pelvic lymph node dissection were performed. Histopathological examination of the lymph nodes revealed dilation of the sinuses and marked increase in the number of histiocytes, which suggested sinus histiocytosis. Sinus histiocytosis is often associated with malignant tumor. It is considered as a sign of the host's immune activation with favorable prognostic significance. Nevertheless, such lymphadenopathy mimics a metastatic lymph node, making proper staging difficult. PMID- 12884875 TI - Unequal access: African-American Medicare beneficiaries and the prescription drug gap. PMID- 12884876 TI - Food labeling: health claims; soluble dietary fiber from certain foods and coronary heart disease. Final rule. AB - The Food and Drug Administration (FDA) is adopting as a final rule, without change, the provisions of the interim final rule that amended the regulation authorizing a health claim on the relationship between beta-glucan soluble fiber from whole oat sources and reduced risk of coronary heart disease (CHD). FDA is taking this action to complete the rulemaking initiated with the interim final rule. PMID- 12884877 TI - Medical devices; obstetrical and gynecological devices; classification of the breast lesion documentation system. Final rule. AB - The Food and Drug Administration (FDA) is classifying the breast lesion documentation system into class II (special controls). The special controls that will apply to this device are discussed later in this document. The agency is taking this action in response to a petition submitted under the Federal Food, Drug, and Cosmetic Act (the act) as amended by the Medical Device Amendments of 1976 (the amendments), the Safe Medical Devices Act of 1990, and the Food and Drug Administration Modernization Act of 1997 (FDAMA). The agency is classifying this device into class II (special controls) in order to provide a reasonable assurance of safety and effectiveness of the device. Elsewhere in this issue of the Federal Register, FDA is publishing a notice of availability of a guidance document that is the special control for this device. PMID- 12884878 TI - Finance issue brief: any willing provider: year end report-2002. AB - Today, almost half the states have laws obligating managed care organizations (MCOs) such as health maintenance organizations (HMOs) and preferred provider organizations (PPOs) to contract with any willing health care provider. Although most provisions are limited to pharmacies or pharmacists, several states have adopted broad provisions applying to hospitals, physicians, chiropractors, pharmacists, podiatrists, therapists and nurses. PMID- 12884879 TI - Finance issue brief: bans on financial incentives: year end report-2002. PMID- 12884880 TI - Finance issue brief: bans on gag clauses: year end report-2002. AB - Forty-eight states prohibit managed care organizations (MCOs) from limiting a provider's ability to discuss potential treatment options with patients. PMID- 12884881 TI - Finance issue brief: comprehensive consumer rights bills: year end report-2002. AB - The health care industry has historically used established formats for patient rights in order to maintain the integrity and quality of care in all settings of care provision. By establishing rights and responsibilities for patients, it increased patient involvement and created boundaries for acceptable practice for health care providers. As the provision of health care is evolving and including the health payor as never before, legislators are faced with the task of bridging the gap for consumers by creating protective measures that are referred to as a "Patient's Bill of Rights". PMID- 12884882 TI - Finance issue brief: consumer grievance procedures: internal and independent appeals: year end report-2002. AB - State lawmakers across the country have begun to play an important role in ensuring that managed care plans establish and maintain grievance and appeals procedures that are responsive to the needs of their constituents. PMID- 12884883 TI - A unitary model of schizophrenia: Bleuler's "fragmented phrene" as schizencephaly. AB - inding a unifying concept behind the diversity of signs and symptoms in schizophrenia is a central challenge to contemporary research. A neo-Bleulerian unitary model is described, which defines the illness as a neurodevelopmentally derived "misconnection syndrome," involving connections between cortical regions and the cerebellum mediated through the thalamus (the cortico-cerebellar-thalamic cortical circuit [CCTCC]). An abnormality in this circuitry, normally used to coordinate both motor and mental activity, leads to misconnections in many aspects of mental activity, or "cognitive dysmetria." As Bleuler originally proposed, "thought disorder" is the primary defining feature of schizophrenia, rather than the more obvious signs and symptoms such as delusions and hallucinations. Cognitive dysmetria, or a disorder in the CCTCC, may provide a heuristic theoretical framework for strategies to explore etiology, pathophysiology, intervention, or prevention. PMID- 12884884 TI - Connecting the "dots" of brain dysfunction in schizophrenia: what does the picture look like? PMID- 12884885 TI - Children with prepubertal-onset major depressive disorder and anxiety grown up. AB - BACKGROUND: The continuity in adulthood of major depressive disorder (MDD) first arising before puberty is largely unknown. This information could guide early treatment and clarify the appropriateness of including children with MDD in genetic studies. METHODS: Eighty-three subjects with onset of MDD, 44 subjects with anxiety disorder and no MDD, and 91 subjects with no evidence of past or current psychiatric disorders were assessed by two psychiatrists before puberty (Tanner stage < III) and were evaluated 10 to 15 years later as adults by an independent team without knowledge of the initial diagnosis. RESULTS: The clinical outcome of children with prepubertal-onset MDD in adulthood includes a high risk of suicide attempts (nearly 3-fold compared with normal controls and 2 fold compared with children with anxiety) and bipolar disorder. Compared with controls, both the children with MDD and those with anxiety went on to have increased risk of substance abuse and conduct disorder but not other disorders, increased use of longterm psychiatric and medical services, and overall impaired functioning. Children with prepubertal-onset MDD with a recurrence of MDD during follow-up had higher rates of MDD in their first-degree relatives. CONCLUSIONS: There is high morbidity in clinically referred children with prepubertal-onset MDD and anxiety, but continuity and specificity of MDD or anxiety disorder in adulthood is less clear. Caution is warranted in selecting clinically referred children with prepubertal-onset MDD for inclusion in genetic studies unless they have a family history of MDD and recurrence of MDD over time. PMID- 12884886 TI - Personality disorders in adolescence and risk of major mental disorders and suicidality during adulthood. AB - BACKGROUND: A community-based longitudinal study was conducted to investigate whether personality disorders (PDs) during adolescence increase the risk for Axis I psychiatric disorders and suicidality during early adulthood. METHOD: Psychosocial and psychiatric interviews were administered to a representative community sample of 717 youths and their mothers from 2 counties in the state of New York in 1975, 1983, 1985-1986, and 1991-1993. Anxiety, disruptive, eating, mood, personality, and substance use disorders and suicidal ideation and behavior were assessed in 1983 and 1985-1986, when the participants were adolescents, and in 1991-1993, when they were young adults. RESULTS: Adolescents with PDs were more than twice as likely as those without PDs to have anxiety, disruptive, mood, and substance use disorders during early adulthood. These associations remained statistically significant after co-occurring Axis I disorders during adolescence were controlled statistically. Cluster A, B, and C PDs and DSM-IV Appendix B PDs during adolescence were all associated with elevated risk for Axis I disorders during early adulthood after co-occurring Axis I and Axis II disorders during adolescence were controlled statistically. Cluster C PDs during adolescence were associated with elevated risk for suicidal ideation or behavior during early adulthood after co-occurring psychiatric disorders and suicidality during adolescence were controlled statistically. CONCLUSIONS: Adolescents in the community with personality disorders are at elevated risk for major mental disorders and suicidal ideation or behavior during early adulthood. This increase in risk is not accounted for by co-occurring Axis I disorders or suicidality during adolescence. PMID- 12884887 TI - Desipramine in opioid-dependent cocaine abusers maintained on buprenorphine vs methadone. AB - BACKGROUND: Cocaine abuse occurs in 40% to 60% of patients entering opioid maintenance treatment, and effective pharmacotherapies are needed for this combined dependence. METHODS: This 13-week, randomized, double-blind, placebo controlled trial evaluated the efficacy of desipramine hydrochloride (0 or 150 mg/d) plus buprenorphine hydrochloride (12 mg/d) or methadone hydrochloride (65 mg/d) in 180 opioid-dependent cocaine abusers (124 men, 56 women). Supervised urine samples were obtained thrice weekly, and self-reported cocaine and heroin use was reported once weekly. Desipramine plasma levels were determined at weeks 4 and 10. RESULTS: In men, opioid abstinence was increased more rapidly over time when treated with methadone than with buprenorphine, whereas cocaine abstinence was increased more with buprenorphine than with methadone. In women, opioid abstinence was increased the least rapidly when treated with buprenorphine plus placebo, while cocaine abstinence was increased more rapidly over time when treated with methadone than with buprenorphine. Regardless of sex or opioid medication, desipramine increased opioid and cocaine abstinence more rapidly over time than placebo. Self-reported opioid use confirmed these findings. Desipramine plasma levels were higher in women than in men, particularly those on buprenorphine maintenance. Higher desipramine plasma levels were associated with greater opioid, but not cocaine, abstinence. CONCLUSIONS: Desipramine may be a useful adjunctive medication in facilitating opioid and cocaine abstinence in opioid-maintained patients. The efficacy of opioid medications to treat opioid or cocaine dependence may differ by sex. These findings highlight the importance of including sex as a factor when examining treatment outcome in these types of trials. PMID- 12884888 TI - Long-term maintenance and discontinuation of imipramine therapy in panic disorder with agoraphobia. AB - BACKGROUND: There has been little systematic work done regarding the long-term treatment of panic disorders. The aim of the present study was to assess the 12 month cumulative risk of relapse specifically due to discontinuation of imipramine and to test the hypothesis that maintenance treatment with imipramine protects patients with panic disorder and agoraphobia from such reversals. METHOD: Following an acute-phase open trial with imipramine (2.25 mg/kg per day) involving 110 patients for 6 months, the 56 patients who were in stable remission, did not require additional treatment, and consented to be randomly assigned to double-blind maintenance (n = 29) or discontinuation (n = 27) conditions were followed up with planned assessments every 2 months during a 1 year period. There were no behaviorally oriented interventions or instructions at any time during the 18 months of the study. RESULTS: Maintenance treatment (1 relapse) and discontinuation (10 relapses) conditions had significantly different survival curves (Mantel-Cox statistic chi(2)1 = 10.47, P = .001). None of the additional 10 variables from demographic, clinical, and open-treatment domains considered in the proportional hazard model disrupted the significant relationship between experimental drug condition and relapse; other things being equal, a patient receiving imipramine maintenance was 92.5% lower in the hazard rate of relapse than a patient receiving placebo. CONCLUSION: The results confirm the very high degree of prophylactic effectiveness of maintenance imipramine treatment and demonstrate that relapse, although substantial, occurs in a minority of patients with panic disorder and agoraphobia who are in stable remission prior to treatment discontinuation. PMID- 12884889 TI - Prevention of relapse in residual depression by cognitive therapy: a controlled trial. AB - BACKGROUND: Previous studies indicate that depressed patients with partial remission and residual symptoms following antidepressant treatment are common and have high rates of relapse. There is evidence that cognitive therapy may reduce relapse rates in depression. METHODS: One hundred fifty-eight patients with recent major depression, partially remitted with antidepressant treatment (mean daily doses equivalent to 185 mg of amitriptyline or 33 mg of fluoxetine) but with residual symptoms of 2 to 18 months' duration, were included in a controlled trial. Subjects were randomized to receive clinical management alone or clinical management plus cognitive therapy for 16 sessions during 20 weeks, with 2 subsequent booster sessions. Subjects were assessed regularly throughout the 20 weeks' treatment and for a further year. They received continuation and maintenance antidepressants at the same dose throughout. RESULTS: Cognitive therapy reduced relapse rates for acute major depression and persistent severe residual symptoms, in both intention to treat and treated per protocol samples. The cumulative relapse rate at 68 weeks was reduced significantly, from 47% in the clinical management control group to 29% with cognitive therapy (hazard ratio 0.54; 95% confidence interval, 0.32-0.93; intention to treat analysis). Cognitive therapy also increased full remission rates at 20 weeks but did not significantly improve symptom ratings. CONCLUSION: In this difficult-to-treat group of patients with residual depression who showed only partial response despite antidepressant treatment, cognitive therapy produced worthwhile benefit. PMID- 12884890 TI - The 4-year course of tic disorders in boys with attention-deficit/hyperactivity disorder. AB - BACKGROUND: Despite long-standing clinical concerns, relatively little is known about the comorbidity between attention-deficit/hyperactivity disorder (ADHD) and tic disorders. Therefore, we examined tic disorders in an ongoing prospective follow-up study of male subjects with ADHD, a sample unselected for any comorbid disorder. METHODS: One hundred twenty-eight male children and adolescents with ADHD and 110 male controls were comprehensively evaluated at baseline and 4 years later. We characterized tic disorders along with a wide range of neuropsychiatric correlates, including other comorbid disorders and indices of psychosocial function in multiple domains (school, cognitive, social, and family). RESULTS: Compared with controls, subjects with ADHD showed more tic disorders at baseline and more new onsets were reported at follow-up. Attention-deficit/hyperactivity disorder and tic disorders appeared to be independent in course: in contrast to low rates of ADHD remission, tic disorders mostly remitted. The age-adjusted rate of ADHD remission was 20% and that of tic remission, 65%. Tic disorders had little effect on the psychosocial functioning of subjects with ADHD. CONCLUSIONS: These findings suggest that comorbidity with a tic disorder has a limited effect on ADHD outcome. However, because of the relatively small sample of subjects with tic disorders, our conclusions should be considered preliminary until confirmed in larger studies of medicated and unmedicated children with ADHD with and without tic disorders. PMID- 12884891 TI - Consensus statement on the upcoming crisis in geriatric mental health: research agenda for the next 2 decades. AB - It is anticipated that the number of people older than 65 years with psychiatric disorders in the United States will increase from about 4 million in 1970 to15 million in 2030. The current health care system serves mentally ill older adults poorly and is unprepared to meet the upcoming crisis in geriatric mental health. We recommend the formulation of a 15- to 25-year plan for research on mental disorders in elderly persons. It should include studies of prevention, translation of findings from bench to bedside, large-scale intervention trials with meaningful outcome measures, and health services research. Innovative strategies are needed to formulate new conceptualizations of psychiatric disorders, especially those given scant attention in the past. New methods of clinical and research training involving specialists, primary care clinicians, and the lay public are warranted. PMID- 12884892 TI - Menopause research: an opportunity for complementary collaborations. PMID- 12884893 TI - Plant models for fundamental research in homeopathy. PMID- 12884894 TI - The homeopathic approach to the treatment of symptoms of oestrogen withdrawal in breast cancer patients. A prospective observational study. AB - This paper reports on an investigation of the homeopathic approach to the management of symptoms of oestrogen withdrawal in women with breast cancer. Forty five patients entered the study. The most common presenting symptoms were hot flushes (HF) (n=38), mood disturbance (n=23), joint pain (n=12), and fatigue (n=16). Other symptoms included sleeplessness, reduced libido, weight gain, cystitis, vaginal dryness and skin eruptions. The active intervention was an individualised homeopathic medicine. Forty women (89%) completed the study. Significant improvements in mean symptom scores were seen over the study period and for the primary end-point 'the effect on daily living' scores. Symptoms other than HF such as fatigue and mood disturbance appear to be helped. Significant improvements in anxiety, depression and quality of life were demonstrated over the study period. The homeopathic approach appears to be clinically useful in the management of oestrogen withdrawal symptoms in women with breast cancer whether on or off Tamoxifen and improves mood disturbance. A placebo-controlled trial would be the next stage in this line of inquiry. PMID- 12884895 TI - Study of the practice of homeopathic general practitioners in France. AB - Little is known about the practice of homeopathic general practitioners (GPs) in France. We report a descriptive, cross-sectional study on this subject. The aim was to acquire a clearer idea of (1) the type of patients who consult homeopathic GPs and (2) the type of treatment provided by such GPs, in terms of diagnoses, prescriptions and costs. The survey was carried out on a representative sample of the French homeopathic GP population and covered three seasons of the year. The data was collected by means of a questionnaire which GPs completed for each consulting patient. It emerged that the type of patients who consult homeopathic GPs are chiefly women, between the ages of 20 and 54, living in an urban environment, not in employment, covered by National Health Insurance for Salaried Workers, and belonging to a Mutual Benefit Association. The most common reasons for consultation were ENT disorders, stress and anxiety. The homeopathic GPs mainly used homeopathy to treat these disorders. On average, four medicinal products per patient were prescribed per 2-month course of treatment, for an average reimbursed cost of 3.78 Euros. PMID- 12884896 TI - Homeopathically prepared gibberellic acid and barley seed germination. AB - The potentisation process by which homeopathic preparations are produced raises the concern that these medicines have placebo effects only, since they theoretically no longer contain active molecules of the diluted substance. Plant models offer a method of examining the efficacy of homeopathically prepared solutions. This study examined the effects of homeopathically prepared gibberellic acid (HGA3) on the germination performance of barley (Hordeum vulgare L.) seeds. The effect of HGA3 (4-200 cH) on seed germination rate and seedling development was compared to that of the most commonly used form of gibberellic acid (GA3), 0.5 g l(-1), and control (distilled water). The extent and type of response was dependent on the vigour level of the seedlot. Treating seeds from three vigour groups in HGA3 consistently resulted in larger seedlings. High vigour seeds treated with HGA3 4, 30 and 200 cH germinated faster, and roots of medium-vigour seedlots treated in HGA3 15 cH were longer. Biphasic effects of HGA3 were also demonstrated. As a plant model, germinating barley seeds successfully demonstrated the ability of HGA3 to produce a biological response. PMID- 12884897 TI - What is the therapeutically active ingredient of homeopathic potencies? AB - The nature of the 'active ingredient', in homeopathic high dilutions is investigated. A model for every degree of dilution is introduced; within this the active ingredient can be dealt with in physical terms. In mathematical terms this model has features which correspond to the axioms of weak quantum theory. Features which are similar to entanglement in ordinary quantum theory are discussed in particular. PMID- 12884898 TI - Patient-practitioner-remedy (PPR) entanglement. Part 3. Refining the quantum metaphor for homeopathy. AB - The notion of patient-practitioner-remedy (PPR) entanglement, previously proposed for homeopathy, is refined by adapting concepts derived from Greenberger, Horne, and Zeilinger's treatment of three-particle entanglement (GHZ states), and a generalised version of quantum theory, called weak quantum theory (WQT). These suggest that for maximum PPR entanglement during the therapeutic encounter, the practitioner's awareness needs to be directed inward as well as outward toward the patient, and that health and disease are mirror images of each other, similar to and represented by, the relationship of complex numbers to their complex conjugates. PMID- 12884899 TI - Saving holistic homeopathic medicine from mechanistic scientism--an urgent need. PMID- 12884900 TI - 20 years ago: British Homoeopathic Journal, July 1983. PMID- 12884901 TI - International conference: Bridging the Credibility Gap; London, 3-4 April 2003. PMID- 12884902 TI - Arnica--a placebo? PMID- 12884903 TI - Witnessing a revolution in voice research: genomics, tissue engineering, biochips and what's next! AB - Due to advances in genomics and tissue engineering, new tools and methods are available for use in voice research. Microarray analysis, a method for examining the expression levels of thousand of genes within 24-48 hours, is a very powerful research tool and has greatly hastened the development of biochips for medical application. The use of microarray analysis in voice research will be discussed and the feasibility of a biochip for voice is presented. Gene expression profiles, a fundamental part of biochip development, are now commonly performed in some voice laboratories. Tissue engineering initiatives have led to the ability to grow and work with laryngeal fibroblasts, Because of the extreme conditions (intense vibration, exposure to inhalants, etc.) that vocal fold fibroblasts tolerate, engineering living lamina propria of vocal folds is complex and challenging. Research efforts to explore these conditions in-vitro and their effect on fibroblast growth is detailed. In summary, applying tools from genomic and tissue engineering fields to vocal science is fruitful and holds great clinical promise. PMID- 12884904 TI - The perception of 'forward' and 'backward placement' of the singing voice. AB - Singing teachers sometimes characterize voice quality in terms of 'forward' and 'backward placement'. In view of traditional knowledge about voice production, it is hard to explain any possible acoustic or articulatory differences between the voices so 'placed'. We have synthesized a number of three-tone melodic excerpts performed by the singing voice. Formant frequencies, and the level and frequency of the singer's formant were varied across the stimuli. Results of a listening test show that the stimuli which were perceived as 'placed forward', correlated not only with higher frequencies of the first and second formants, but also with the higher frequency and level of the singer's formant. PMID- 12884905 TI - Methods to study pre-school teachers' voice at work: simultaneous recordings with a voice accumulator and a DAT recorder. AB - Long-term recordings with reliable methods are desirable for objective documentation of voice use during natural conditions. The purpose of this study was to evaluate a voice accumulator (VAC) with a digital audiotape (DAT) recorder as a reference. The VAC is based on a microprocessor that accumulates information about fundamental frequency (F0) and phonation time. A contact microphone attached to the front of the neck registers vocal fold vibrations. The DAT recorder was connected to two microphones for airborne signals placed at equal distance from the mouth close to the subject's ears. The computer program Aura was used to separate the subject's voice from the background noise. The Soundswell program was used for F0 and phonation time analysis. Two tests were performed: 1) One female speech-language pathologist was recorded with the two devices simultaneously in a sound-proof booth. She read a standard text with different voice qualities and sustained vowels with increasing F0 and intensity separately. The results showed good agreement between the two methods with respect to F0 and phonation time. However, the VAC failed to register high frequencies above around 440 Hz as well as low intensities. 2) Three female pre school teachers were recorded with the two devices simultaneously during a working day. Results showed high correlations between the two methods in terms of long-term measurements of F0 and phonation time for two subjects For one subject with subcutaneous soft tissue on the neck, the registration with the contact microphone was not reliable. It was concluded that the VAC has potential for assessment of occupational voice disorders if certain limitations of the method are considered. PMID- 12884906 TI - The crashed voice--a potential for change: a psychotherapeutic view. AB - Jungian analysis is based on the work and extensive writings of C.G. Jung. Jung worked for a while with Freud who expected him to carry on the work of psychoanalysis. However, the two men were very different and Jung broke away from the Freudian model to develop his own unique view of the unconscious and to explore what he believed to be its innate capacity for healing. He said: "My life is the story of the self realization of the unconscious. Everything in the unconscious seeks outward manifestation, and the personality too desires to evolve out of its unconscious conditions and to experience itself as a whole." (Memories, Dreams, Reflections C.G. Jung. 1963) The goal of the therapeutic work for Jung was individuation--the never ending process of becoming fully oneself. This process represents the individual's capacity to choose creatively between both internal and external necessity without denying either. Jungian analysts work in some similar ways to other analysts and see dreams as central to this process. They also sometimes work with archetypal symbols and other forms of creative expression such as painting or dance. All the methods used are there to enable patients to explore their unconscious, understand and accept who they are, and access powers for healing the self. People in creative and expressive fields are often drawn to the work because of its belief in the uniqueness of the individual and its understanding of the healing power of a creative relationship to oneself. PMID- 12884907 TI - Epidermal growth factor receptor family tyrosine kinases as signal integrators and therapeutic targets. AB - Activating mutations, gene amplification and overexpression of human epidermal growth factor receptor (erbB or HER) family kinases have been implicated as integral contributors to a variety of cancers. However, there is increasing evidence that the HER family receptors propagate not only signals initiated by their own ligands but also those initiated by multiple other signaling pathways. Attenuation of HER family signaling is a developing strategy for the management of human malignancies and is the subject of a number of ongoing clinical trials. The purpose of this review is to provide a timely update of recent reports in the rapidly evolving area of HER family biology and the emerging integral role of these receptors in malignant transformation and as targets of cancer therapy. PMID- 12884908 TI - c-Met: structure, functions and potential for therapeutic inhibition. AB - Studies on signal transduction pathways have generated various promising molecular targets for therapeutic inhibition in cancer therapy. Receptor tyrosine kinases represent an important class of such therapeutic targets. c-Met is a receptor tyrosine kinase that has been shown to be overexpressed and/or mutated in a variety of malignancies. A number of c-Met activating mutations, many of which are located in the tyrosine kinase domain, have been detected in various solid tumors and have been implicated in invasion and metastasis of tumor cells. It is known that stimulation of c-Met via its natural ligand, hepatocyte growth factor (also known as scatter factor, HGF/SF) results in a plethora of biological and biochemical effects in the cell. Activation of c-Met signaling can lead to scattering, angiogenesis, proliferation, enhanced cell motility, invasion, and eventual metastasis. In this review, the role of c-Met dysregulation in tumor progression and metastasis is discussed in detail with particular emphasis on c Met mutations. Moreover, we summarize current knowledge on various pathways of c Met signal transduction, highlighting the central role in the cytoskeletal functions. In this summary is included recent data in our laboratory indicating that phosphorylation of focal adhesion proteins, such as paxillin, p125FAK, and PYK2, occurs in response to c-Met stimulation in lung cancer cells. Most importantly, current data on c-Met suggest that when mutated or overexpressed in malignant cells, c-Met would serve as an important therapeutic target. PMID- 12884909 TI - The type-1 insulin-like growth factor receptor tyrosine kinase and breast cancer: biology and therapeutic relevance. AB - The development of the mammary gland requires the coordinated expression of hormones and growth factors. Likewise, some transformed breast cells continue to respond to these same extracellular signals. Thus, understanding the mechanisms that control normal development of tissues can lead to new therapeutic targets. The insulin-like growth factor (IGF) system plays an important role in the normal development and function of the mammary gland. Accumulating evidence suggests that the IGFs are also key regulators of the malignant phenotype. The IGFs stimulate proliferation, promote survival, and enhance metastatic potential of breast cancer cells. Although multiple receptors for the IGFs have been identified, the IGFs primarily exert their biologic effects through ligation of the type I IGF receptor tyrosine kinase (IGF1R). IGF binding to the IGF1R initiates an intracellular signaling cascade that leads to changes in gene expression and cell biology. This review will focus on the evidence that the IGF1R is a relevant treatment target in breast cancer. PMID- 12884910 TI - Src family kinases in tumor progression and metastasis. AB - The Src family of non-receptor protein tyrosine kinases plays critical roles in a variety of cellular signal transduction pathways, regulating such diverse processes as cell division, motility, adhesion, angiogenesis, and survival. Constitutively activated variants of Src family kinases, including the viral oncoproteins v-Src and v-Yes, are capable of inducing malignant transformation of a variety of cell types. Src family kinases, most notably although not exclusively c-Src, are frequently overexpressed and/or aberrantly activated in a variety of epithelial and non-epithelial cancers. Activation is very common in colorectal and breast cancers, and somewhat less frequent in melanomas, ovarian cancer, gastric cancer, head and neck cancers, pancreatic cancer, lung cancer, brain cancers, and blood cancers. Further, the extent of increased Src family activity often correlates with malignant potential and patient survival. Activation of Src family kinases in human cancers may occur through a variety of mechanisms and is frequently a critical event in tumor progression. Exactly how Src family kinases contribute to individual tumors remains to be defined completely, however they appear to be important for multiple aspects of tumor progression, including proliferation, disruption of cell/cell contacts, migration, invasiveness, resistance to apoptosis, and angiogenesis. This review details the evidence for Src family activation in human tumors, and emphasizes possible consequences to tumor progression. Given the ability of Src and its family members to participate in so many aspects of tumor progression and metastasis, Src family kinases are attractive targets for future anti-cancer therapeutics. PMID- 12884911 TI - FAK regulates biological processes important for the pathogenesis of cancer. AB - Since its initial discovery as a substrate and binding partner for the Src oncogene, a role for the focal adhesion kinase (FAK) in cancer has been speculated. In this review the clinical evidence correlating FAK overexpression with cancer and the experimental evidence demonstrating that FAK can control some phenotypes associated with cancer will be discussed. In addition, the emerging theme of interactions between the FAK and growth factor signaling pathways will be described. The evidence presented in this review provides a compelling case for a role for FAK in the pathology of human cancer. PMID- 12884912 TI - The role of integrin-linked kinase (ILK) in cancer progression. AB - Integrin-linked kinase (ILK) is an intracellular protein, which interacts with the cytoplasmic domains of integrin beta and beta3 subunits. ILK is a 59 kDa protein containing a phosphoinositide phospholipid-binding domain flanked by an N terminal ankyrin repeat domain and a C-terminal serine/threonine protein kinase domain. Genetic and biochemical evidence have established an essential role of ILK in connecting integrins to the actin cytoskeleton. Apart from integrins, ILK interacts with several adaptor and signaling proteins resulting in its activation and localization to focal adhesion plaques. The kinase activity of ILK is stimulated upon integrin engagement, as well as by growth factors and chemokines in a PI-3Kinase-dependent manner. ILK can mediate the phosphorylation of a variety of intracellular substrates, most notable of which are: protein kinase B (PKB/Akt), glycogen synthase kinase-3 (GSK-3) and myosin light chain. Gain and loss of function strategies have shown that overexpression, and/or constitutive activation of ILK results in oncogenic transformation and progression to invasive and metastatic phenotypes. In addition ILK expression and activity are upregulated in several types of cancers. In this review, we summarize the adaptor and signaling properties ofILK, and also progress in the identification of therapeutic strategies for inhibition of ILK activity. PMID- 12884913 TI - P21-activated kinases in human cancer. AB - A balance between proliferation, differentiation, migration and death of cells is critical for the normal development of an organism. Perturbations of this balance can contribute to cancer development. The p21-activated serine/threonine kinases (Paks) play an important role in a variety of cellular functions including cell morphogenesis, motility, survival, angiogenesis, and mitosis. Paks were initially identified as an effector molecules of RHO GTPases; however, recent evidence that they can be activated in both GTPase-dependent and -independent manners expands our understanding of their physiologic functions. Paks play an important role in growth factor signaling, leading to cytoskeletal reorganization that subsequently influences growth factor-mediated cell migration and metastasis functions. Recent findings that Paks play a role in mitosis, nuclear receptor-signaling and deregulation of Pak in cancer cells suggest that these kinases play an important role in both normal development and cancer progression. In this review, we summarize the results of recent advances into the role of Paks in tumorigenesis and metastasis. PMID- 12884914 TI - Role of MAP kinase in tumor progression and invasion. AB - Activation of the mitogen-activated protein kinase/extracellular signal-regulated kinase (MAPK/ERK) pathway is a frequent event in tumorigenesis. MAPKs have been implicated in cell migration, proteinase-induction, regulation of apoptosis, and angiogenesis, events that are essential for successful completion of metastasis. In this review, we discuss the potential role that MAPKs play in metastasis by regulating cell migration, proteinase-induction and apoptosis. PMID- 12884915 TI - Deregulation of NF-kappaB and its upstream kinases in cancer. AB - The transcription factor NF-kappaB is known for its function in regulating immune and inflammatory responses. However, recent evidence suggests that NF-kappaB also plays a pivotal role in controlling cell proliferation, apoptosis, and cell migration. Deregulated activation of NF-kappaB has been observed in various cancers. Over the past few years. significant progress has been made to elucidate the mechanisms of NF-kappaB activation in both normal and cancer cells. Notably, a large number of protein kinases have been shown to stimulate NF-kappaB activity under different conditions, and some of these kinases are aberrantly activated in cancer cells. In this review, we discuss our current knowledge of NF-kappaB activation, with a focus on the NF-kappaB-activating kinases and their roles in cancer formation. PMID- 12884916 TI - Jak family of kinases in cancer. AB - The family of Jak kinases is composed from at least four different tyrosine kinases (Tyk2, Jak1, Jak2, Jak3) that share significant structural homology with each other. The members of this family of kinases associate constitutively with a variety of cytokine and hormone receptors. Upon binding of the specific ligands to their receptors, Jak kinases are rapidly activated and their kinase activities induced, to regulate tyrosine phosphorylation of various effectors and initiate activation of downstream signaling pathways. The discovery of this family of tyrosine kinases dates back in the early 1990s with the cloning of the Tyk-2 tyrosine kinase as a critical element of the Type I interferon signaling pathway. Extensive work over the last few years has provided evidence that Jak kinases play important roles in the generation of responses for interferons, which are cytokines that exhibit important antitumor activities. There is also accumulating evidence that constitutive activation of different Jaks and Stats mediates neoplastic transformation and promotes abnormal cell proliferation in various malignancies. This review summarizes the role of various Jak-kinase dependent pathways in malignancies and discusses the therapeutic implications of the recent advances in the field. PMID- 12884917 TI - Regulators of G1 cyclin-dependent kinases and cancers. AB - The mammalian cell cycle can be divided into four phases: G1 (gap phase 1), S (DNA synthesis), G2 (gap phase 2), and M (mitosis). Progression through each phase of the cell cycle is delicately controled by the activity of different cyclin-dependent kinases (CDKs) and their regulatory subunits known as cyclins. CDK2, CDK4, CDK6 and their associated cyclins control the G1 to S phase transition. The association of CDK4 or CDK6 with D-type cyclins is critical for G1 phase progression, whereas the CDK2-cyclin E complex is important for initiation of the S phase. Cancer can originate from dysregulation of these regulators. A variety of intrinsic and extrinsic signals were recently identified to regulate these G1 or G1/S CDKs and cyclins. Here we discuss the regulators of these protein kinases at different mechanistic level with a hope that these insights can be applied to develop therapeutic strategies for cancer treatment. PMID- 12884918 TI - The Aurora kinases: role in cell transformation and tumorigenesis. AB - Aurora kinases representing a novel family of serine/threonine kinases have been identified as key regulators of the mitotic cell division process. The three members of this kinase family, identified so far, referred to as Aurora-A, Aurora B and Aurora-C kinases, are close homologues of the prototypic yeast Ipll and Drosophila aurora kinases, which are known to be involved in the regulation of centrosome function, bipolar spindle assembly and chromosome segregation processes. All three members of the mammalian kinase family have a catalytic domain that is highly conserved with a short C-terminal domain and an N-terminal domain of varying sizes. Following their discovery about five years ago, extensive research has focused on understanding the biological roles of these kinases and elucidation of their pathways, which regulate cell proliferation and maintenance of normal cellular phenotypes. Significant interest in the subject was generated since all three Aurora kinases family members were reported to be overexpressed in many human cancers, and elevated expression has been correlated with chromosomal instability and clinically aggressive disease in some instances. Ectopic overexpression of one member of the family, Aurora-A, was shown to induce oncogenic transformation in cells. Unlike most other putative oncogenes identified, so far, members of this kinase family are expressed and active at the highest level during G2-M phase of the cell cycle. Aurora kinases are localized at the centrosomes of interphase cells, at the poles of the bipolar spindle and in the midbody of the mitotic apparatus. Substrates identified for the Aurora-A and Aurora-B kinases, include a kinesin-like motor protein, spindle apparatus proteins, histone H3 protein, kinetochore protein and the tumor suppressor protein p53. Identification of Aurora kinases as RasGAP Src homology 3 domain binding protein, also implicates these kinases as potential effectors in the Ras pathway relevant to oncogenesis. Abnormal elevated expression of Aurora kinases detected in human cancer cells could help explain the underlying biological mechanisms responsible for the development of many cellular phenotypes associated with malignant cells. Identification of these mechanisms offers the possibility of designing novel targeted therapies for cancer in the future. PMID- 12884919 TI - Intraperitoneal cancer dissemination: mechanisms of the patterns of spread. AB - BACKGROUND: Well known patterns govern the distribution of hematogenous and lymphatic metastasis of cancer. In the past the distribution of cancer cells free within abdominal cavity received little attention and was thought to be a random event. However, surgical observation led the authors to generate and test hypotheses regarding patterns of spread that vary with tumor type, with the intraperitoneal environment, and with the physiology of the peritoneal surface tissues. METHODS: The distribution and volume of peritoneal surface malignancy was prospectively recorded in 129 patients with 5 different types of tumors at the time of cytoreductive surgery. The malignancies studied included pseudomyxoma peritonei (PMP) of appendiceal origin, colonic mucinous adenocarcinoma (MA), nonmucinous colonic adenocarcinoma (NMA), ovarian carcinoma (OV) and sarcoma (SA). The abdominal and pelvic cavity was divided into 3 horizontal sectors, 9 regions and 25 sites. The incidences of tumor implants in these designated areas were statistically analyzed for each tumor type and comparisons between tumor types studied. RESULTS: The magnitude of intraperitoneal cancer dissemination was similar for mucinous tumors, including PMP and MA and significantly higher than for non-mucinous tumors. Also the mucinous cancers were more likely to be present in the upper horizontal sector than were non-mucinous. When NMA was compared to PMP and MA the epigastric region was significantly less likely to contain tumor. For all cancer diagnoses the colon, greater omentum and cul-de-sac of Douglas were most often affected. The ileocecal valve region was more likely to have large tumor masses on its surface than small bowel surface or small bowel mesentery. CONCLUSIONS: Peritoneal carcinomatosis had a wider distribution when mucinous fluid was present; this cancer distribution by intraperitoneal fluid hydrodynamics occurred regardless of histologic aggressiveness. The organs that have peritoneal fluid resorption (omentum and omental appendages) have a high incidence of implants. Small bowel and its mesentery free to move by peristalsis had a reduced incidence of implants as compared to the ileocecal area, which is fixed to the retroperitoneum. These observations may facilitate efforts to treat peritoneal surface malignancy. PMID- 12884920 TI - Medical applications of synchrotron radiation. AB - The medical imaging and therapeutic technologies that are based on the use of radiation are reviewed briefly, with special emphasis on the recent developments of synchrotron radiation (SR) methods. New results have been achieved in all of these areas since the last comprehensive reviews were written in this field. This topical review is intended to make the latest possible results and complete set of references available. The different contrast mechanisms in imaging by x-rays are described. The applications range from whole-body imaging to studies of atomic and molecular structures. The SR imaging applications include coronary angiography, bronchography, mammography, computed tomography, x-ray microscopy and imaging by scattering. The therapy applications include photon activation therapy and microbeam radiation therapy. PMID- 12884921 TI - Estimation of parameters of dose-volume models and their confidence limits. AB - Predictions of the normal-tissue complication probability (NTCP) for the ranking of treatment plans are based on fits of dose-volume models to clinical and/or experimental data. In the literature several different fit methods are used. In this work frequently used methods and techniques to fit NTCP models to dose response data for establishing dose-volume effects, are discussed. The techniques are tested for their usability with dose-volume data and NTCP models. Different methods to estimate the confidence intervals of the model parameters are part of this study. From a critical-volume (CV) model with biologically realistic parameters a primary dataset was generated, serving as the reference for this study and describable by the NTCP model. The CV model was fitted to this dataset. From the resulting parameters and the CV model, 1000 secondary datasets were generated by Monte Carlo simulation. All secondary datasets were fitted to obtain 1000 parameter sets of the CV model. Thus the 'real' spread in fit results due to statistical spreading in the data is obtained and has been compared with estimates of the confidence intervals obtained by different methods applied to the primary dataset. The confidence limits of the parameters of one dataset were estimated using the methods, employing the covariance matrix, the jackknife method and directly from the likelihood landscape. These results were compared with the spread of the parameters, obtained from the secondary parameter sets. For the estimation of confidence intervals on NTCP predictions, three methods were tested. Firstly, propagation of errors using the covariance matrix was used. Secondly, the meaning of the width of a bundle of curves that resulted from parameters that were within the one standard deviation region in the likelihood space was investigated. Thirdly, many parameter sets and their likelihood were used to create a likelihood-weighted probability distribution of the NTCP. It is concluded that for the type of dose response data used here, only a full likelihood analysis will produce reliable results. The often-used approximations, such as the usage of the covariance matrix, produce inconsistent confidence limits on both the parameter sets and the resulting NTCP values. PMID- 12884922 TI - Characterization of the water-equivalent material WTe for use in electron beam dosimetry. AB - This paper describes the characterization of the water-equivalent material WTe (produced by St Bartholomew's Hospital, London). The use of epoxy resin phantoms offers a number of advantages over water for radiotherapy dosimetry in terms of robustness and ease of use, but the published uncertainties in the fluence corrections for such phantoms significantly increase the overall uncertainty in the measurement of absorbed dose to water at the reference point. Depth ionization data were obtained in water and WTe for electron beams in the range 4 MeV to 16 MeV and it was found that the measured fluence in the WTe phantom was approximately 0.4% higher than in a water phantom at the same depth. For measurements only at the reference depth this difference was less, with the fluence in the WTe phantom being 0.2% higher. The standard uncertainty on this value is estimated to be +/- 0.12%, which represents a significant improvement over previous measurements. It was also found that the range scaling factor is not equal to unity, as previously recommended for this material, but that the data was best fitted by the relation 1 mm WTe = 1.01 mm water (with an uncertainty of +/- 0.2%). The results obtained confirm previous investigations of WTe as to its suitability for reference ion chamber dosimetry in the radiotherapy clinic. However, the recommendation is still to use a water phantom wherever possible. PMID- 12884923 TI - The value of radiochromic film dosimetry around air cavities: experimental results and Monte Carlo simulations. AB - In this study we investigate radiochromic film dosimetry around air cavities with particular focus on the perturbation of the dose distribution by the film when the film is parallel to the beam axis. We considered a layered polystyrene phantom containing an air cavity as a model for the air-soft tissue geometry that may occur after surgical resection of a paranasal sinus tumour. A radiochromic film type MD-55 was positioned within the phantom so that it intersected the cavity. Two phantom set-ups were examined. In the first case, the air cavity is at the centre of the phantom, thus the film is lying along the central beam axis. In the second case, the cavity and film are located 2 cm offset from the phantom centre and the central beam axis. In order to examine the influence of the film on the dose distribution and to interpret the film-measured results, Monte Carlo simulations were performed. The film was modelled rigorously to incorporate the composition and structure of the film. Two field configurations, a 1 x 10 cm2 field and a 10 x 10 cm2 field, were examined. The dose behind the air cavity is reduced by 6 to 7% for both field configurations when a film that intersects the cavity contains the central beam axis. This is due to the attenuation exerted by the film when photons cross the cavity. Offsetting the beam to the cavity and the film by 2 cm removes the dose reduction behind the air cavity completely. Another result was that the rebuild-up behind the cavity for the 10 x 10 cm2 field, albeit less significant than for the 1 x 10 cm2 field, could only be measured by the film that was placed offset with respect to the central beam axis. Although radiochromic film is approximately soft-tissue equivalent and energy independent as compared to radiographic films, care should be taken in the case of inhomogeneous phantoms when the film intersects air cavities and contains the beam central axis. Errors in dose measurement can be expected distal to the air cavity due to attenuation in the film itself. This attenuation would not occur in the absence of the film. Both experiments and Monte Carlo computations support this conclusion. PMID- 12884924 TI - Modelling the elastic scattering in diagnostic radiology: the importance of structure form factors. AB - The importance of structure form factors in describing elastic scattering in diagnostic radiology was studied through a Monte Carlo code built to reproduce scattering in large water samples. The code, developed by us, considers all relevant interactions, including multiple scattering and interference due to scattering by the liquid structure. Geometrical conditions and energies similar to those found in radiology were used. The secondary to primary radiation ratio using the usual free atom approximation and the structure form factor was obtained and both approaches were compared. Calculations of radiological parameters such as the angular distribution of photons incident on the detector and the fraction of scattered photons stopped by anti-scattering grids were also performed considering mammography, thorax and abdomen radiography conditions. The results have shown that S(beta)/P depends on the experimental set-up, being more important for low momentum transfers and sample sizes for which the multiple scattering is not expected to be significant, as in the case of mammography. It was also verified that large samples increase the probability of multiple scattering, masking the structure peak in S(beta) and making the sample structure important just for relatively thin samples. Considering mammography-like geometry, the maximum of the S(beta)/P distribution considering structure form factors occurs around 15 degrees while the correspondent maximum without considering the structure factors occurs around 10 degrees for any sample thickness. S(beta)/P is almost independent of the irradiation field, with the maximum remaining at 15 degrees and 10 degrees for the SFF and FAFF, respectively. The cases studied in this paper stress some conditions in which it is mandatory to use SFF, but since it requires no further significant efforts, the SFF approach is recommended as a standard procedure when describing the elastic scattering process in radiology. PMID- 12884925 TI - Sliding window dual gradient echo (SW-dGRE): T1 and proton resonance frequency (PRF) calibration for temperature imaging in polyacrylamide gel. AB - The aim of the work is to evaluate a magnetic resonance imaging (MRI) thermometry sequence suitable for targeting of focused ultrasound (FUS) when used in vascular occlusion studies. A sliding window dual gradient echo (SW-dGRE) sequence was used. This sequence has the capability of monitoring both T1 relaxation and phase changes, which vary with temperature. Preliminary work involved quantification of the changes in T1 relaxation time with temperature and obtaining the PRF shift coefficient in polyacrylamide gel as it underwent an exothermic reaction during polymerization (avoiding the use of an external heat source). Temperature changes were visualized using thermal maps acquired with the sequence. For FUS guidance a thermal imaging technique is required with a temporal resolution <5 s, a spatial resolution of approximately 1 mm and a temperature resolution of approximately 5 degrees C. The sequence was optimized to improve the CNR (contrast to noise ratio) and SNR (signal to noise ratio) in the phase and magnitude images respectively. The PRF coefficient obtained for the polyacrylamide gel was -9.98 +/- 0.24 ppb degrees C(-1), whilst deltaT1 and temperature change were related by a proportionality factor, the T1 temperature coefficient, of 102.3 +/- 2.9 ms degrees C(-1). The sequence produces an image at every 1.4 s interval. In both magnitude and phase data, the in-plane resolution is +/- 1.2 mm and the temperature resolution is approximately 2 degrees C. The advantage of this sequence is that the temperature obtained from the magnitude data can be confirmed independently using the phase data and vice versa. Thus the sequence can essentially be crosschecked. PMID- 12884926 TI - Optimization of helical tomotherapy treatment plans for prostate cancer. AB - Helical tomotherapy (HT) is a novel treatment approach where the ring gantry irradiation geometry of a helical CT scanner is combined with an intensity modulated megavoltage x-ray fan beam. An inverse treatment planning system (TomoTherapy Inc., Madison) was used to optimize the treatment plans for ten randomly selected prostate patients. Five different sets of margins (2, 5, 7.5 and 10 mm uniform 3D margins and a non-uniform margin of 5 to 10 mm) were employed for the prostate (GTV2) and seminal vesicles (GTV1). The dose distribution was evaluated in targets, rectum, bladder and femoral heads. HT plans are characterized by a rapid dose fall off around the target in all directions resulting in low doses (less than 30% of the dose at ICRU reference point) to the femurs in all cases. Up to a margin of 5 mm for target structures, it was always possible to satisfy the requirements for dose delivery set by RTOG protocol P-0126. Using a 'class solution', HT plans require minimal operator interaction and result in excellent sparing of normal structures in prostate radiotherapy. PMID- 12884927 TI - Dual-energy imaging in full-field digital mammography: a phantom study. AB - A dual-energy technique which employs the basis decomposition method is being investigated for application to digital mammography. A three-component phantom, made up of plexiglas, polyethylene, and water, was doubly exposed with the full field digital mammography system manufactured by General Electric. The 'low' and 'high' energy images were recorded with a Mo/Mo anode-filter combination and a Rh/Rh combination, respectively. The total dose was kept within the acceptable levels of conventional mammography. The first hybrid images obtained with the dual-energy algorithm are presented in comparison with a conventional radiograph of the phantom. Image-quality characteristics at contrast cancellation angles between plexiglas and water are discussed. Preliminary results show that a combination of a standard Mo-anode 28 kV radiograph with a Rh-anode 49 kV radiograph provides the best compromise between image-quality and dose in the hybrid image. PMID- 12884929 TI - Three-dimensional forward solver and its performance analysis for magnetic resonance electrical impedance tomography (MREIT) using recessed electrodes. AB - In magnetic resonance electrical impedance tomography (MREIT), we try to reconstruct a cross-sectional resistivity (or conductivity) image of a subject. When we inject a current through surface electrodes, it generates a magnetic field. Using a magnetic resonance imaging (MRI) scanner, we can obtain the induced magnetic flux density from MR phase images of the subject. We use recessed electrodes to avoid undesirable artefacts near electrodes in measuring magnetic flux densities. An MREIT image reconstruction algorithm produces cross sectional resistivity images utilizing the measured internal magnetic flux density in addition to boundary voltage data. In order to develop such an image reconstruction algorithm, we need a three-dimensional forward solver. Given injection currents as boundary conditions, the forward solver described in this paper computes voltage and current density distributions using the finite element method (FEM). Then, it calculates the magnetic flux density within the subject using the Biot-Savart law and FEM. The performance of the forward solver is analysed and found to be enough for use in MREIT for resistivity image reconstructions and also experimental designs and validations. The forward solver may find other applications where one needs to compute voltage, current density and magnetic flux density distributions all within a volume conductor. PMID- 12884928 TI - Accelerated median root prior reconstruction for pinhole single-photon emission tomography (SPET). AB - Pinhole collimation can be used to improve spatial resolution in SPET. However, the resolution improvement is achieved at the cost of reduced sensitivity, which leads to projection images with poor statistics. Images reconstructed from these projections using the maximum likelihood expectation maximization (ML-EM) algorithms, which have been used to reduce the artefacts generated by the filtered backprojection (FBP) based reconstruction, suffer from noise/bias trade off: noise contaminates the images at high iteration numbers, whereas early abortion of the algorithm produces images that are excessively smooth and biased towards the initial estimate of the algorithm. To limit the noise accumulation we propose the use of the pinhole median root prior (PH-MRP) reconstruction algorithm. MRP is a Bayesian reconstruction method that has already been used in PET imaging and shown to possess good noise reduction and edge preservation properties. In this study the PH-MRP algorithm was accelerated with the ordered subsets (OS) procedure and compared to the FBP, OS-EM and conventional Bayesian reconstruction methods in terms of noise reduction, quantitative accuracy, edge preservation and visual quality. The results showed that the accelerated PH-MRP algorithm was very robust. It provided visually pleasing images with lower noise level than the FBP or OS-EM and with smaller bias and sharper edges than the conventional Bayesian methods. PMID- 12884930 TI - Post-processing noise removal algorithm for magnetic resonance imaging based on edge detection and wavelet analysis. AB - A post-processing noise suppression technique for biomedical MRI images is presented. The described procedure recovers both sharp edges and smooth surfaces from a given noisy MRI image; it does not blur the edges and does not introduce spikes or other artefacts. The fine details of the image are also preserved. The proposed algorithm first extracts the edges from the original image and then performs noise reduction by using a wavelet de-noise method. After the application of the wavelet method, the edges are restored to the filtered image. The result is the original image with less noise, fine detail and sharp edges. Edge extraction is performed by using an algorithm based on Sobel operators. The wavelet de-noise method is based on the calculation of the correlation factor between wavelet coefficients belonging to different scales. The algorithm was tested on several MRI images and, as an example of its application, we report the results obtained from a spin echo (multi echo) MRI image of a human wrist collected with a low field experimental scanner (the signal-to-noise ratio, SNR, of the experimental image was 12). Other filtering operations have been performed after the addition of white noise on both channels of the experimental image, before the magnitude calculation. The results at SNR = 7, SNR = 5 and SNR = 3 are also reported. For SNR values between 5 and 12, the improvement in SNR was substantial and the fine details were preserved, the edges were not blurred and no spikes or other artefacts were evident, demonstrating the good performances of our method. At very low SNR (SNR = 3) our result is worse than that obtained by a simpler filtering procedure. PMID- 12884931 TI - High-resolution EEG mapping: an equivalent charge-layer approach. AB - Brain electrical signal is one of the windows to understanding neural activities. Various high-resolution imaging techniques have been developed to reveal the electrical activities underneath the cortical surface from scalp electroencephalographic recordings, such as scalp Laplacian, cortical surface potential, equivalent charge layer (ECL) and equivalent dipole layer (EDL). In this work, we develop forward density formulae for the ECL and the EDL of neural electric sources in a 4-concentric-sphere head model, and compare ECL with EDL in theory, simulation and real evoked data tests. The results confirm that the ECL map may be of higher spatial resolution than the EDL map. PMID- 12884932 TI - An alternative method for determination of low-frequency specific absorption rate patterns in homogeneous phantoms. AB - We propose a new application of voltage gradient measurements to determine specific absorption rate (SAR) at low frequencies where quasi-static electromagnetic conditions apply. This method, which we call the voltage gradient method, relies on direct measurement of the voltage field rather than measurement of the electric field or thermal transients. The voltage gradient method is fast and can be implemented with voltmeters of moderate cost. We tested the voltage gradient method using normal saline, in a phantom with simple geometry, and a sine wave voltage source at 5, 10, 20 and 50 kHz. Compared to the SAR measured thermally in the same phantom, the voltage gradient method produced almost identical curves when normalized. When the results of the voltage gradient method were scaled to the same power level used for the thermal SAR, the agreement was compatible with typical thermal SAR accuracy. PMID- 12884933 TI - The use of Fourier-transform infrared spectroscopy for the quantitative determination of glucose concentration in whole blood. AB - Fourier-transform infrared transmission spectroscopy has been used for the determination of glucose concentration in whole blood samples from 28 patients. A 4-vector partial least-squares calibration model, using the spectral range 950 1200 cm(-1), yielded a standard-error-of-prediction of 0.59 mM for an independent test set. For blood samples from a single patient, we found that the glucose concentration was proportional to the difference between the values of the second derivative spectrum at 1082 cm(-1) and 1093 cm(-1). This indicates that spectroscopy at these two specific wavenumbers alone could be used to determine the glucose concentration in blood plasma samples from a single patient, with a prediction error of 0.95 mM. PMID- 12884934 TI - Calculating the uncertainty in lead concentration for in vivo bone lead x-ray fluorescence. AB - A revised mathematical treatment of the calibration line intercept has been published for in vivo bone lead measurements using 109Cd-based K-shell x-ray fluorescence. The revised calibration line treatment prompts changes, presented herein, to the method for calculating the measurement uncertainty. PMID- 12884935 TI - Effects of 0.075 Gy x-ray irradiation on the expression of IL-10 and IL-12 in mice. AB - The objective was to observe the effects of 0.075 Gy low dose radiation (LDR) on the synthesis of IL-10 in splenocytes and the secretion of IL-12 by peritoneal macrophages. Kunming mice were selected and randomly grouped. Northern blot and flow cytometry were adopted to detect the changes of IL-10 at mRNA and protein levels, respectively. Northern blot and ELISA were used, respectively, to examine the changes of IL-12 p35/p40 mRNA and IL-12 p70 protein levels. IL-10 mRNA decreased significantly in splenocytes, while both IL-12 p35 and p40 subunit mRNA levels in macrophages increased after whole body irradiation (WBI) with 0.075 Gy x-rays. Meanwhile, IL-10 synthesis in splenocytes decreased beginning from 4 h after exposure and remaining at a lower level up to 48 h, and IL-12 secretion by macrophages was found to take the opposite direction, i.e. increase significantly. In conclusion, WBI with 0.075 Gy x-rays may suppress IL-10 both at the mRNA level and protein level and stimulate IL-12 expression simultaneously, which might contribute to a shift of the immune response in favour of Th1 differentiation. PMID- 12884936 TI - Image analysis can be used to detect spatial changes in the histopathology of pancreatic tumours. AB - Pancreatic cancer is frequently associated with intense growth of fibrous tissue at the periphery of tumours, but the histopathological quantification of this stromal reaction has not yet been used as a prognostic factor because of the difficulty of obtaining quantitative measures using manual methods. Manual histological grading is a poor indicator of outcome in this type of cancer and there is a clinical need to establish a more sensitive indicator. Recent pancreatic tumour biology research has focused upon the stromal reaction and there is an indication that its histopathological quantification may lead to a new prognostic indicator. Histological samples from 21 cases of pancreatic carcinoma were stained using the sirius red, light-green method. Multiple images from the centre and periphery of each tumour were automatically segmented using colour cluster analysis to subdivide each image into representative colours. These were classified manually as stroma, cell cytoplasm or lumen in order to measure the area of each component in each image. Measured areas were analysed to determine whether the technique could detect spatial differences in the area of each tissue component over all samples, and within individual samples. Over all 21 cases, the area of stromal tissue at the periphery of the tumours exceeded that at the centre by an average of 10.0 percentage points (P < 0.001). Within individual tumours, the algorithm was able to detect significantly more stroma (P < 0.05) at the periphery than the centre in 11 cases, whilst none of the remaining cases had significantly more stromal tissue at the centre than the periphery. The results demonstrate that semi-automated analysis can be used to detect spatial differences in the area of fibrous tissue in routinely stained sections of pancreatic cancer. PMID- 12884937 TI - Summary of the role of statins in the treatment of dyslipidemia. PMID- 12884938 TI - Weight of the evidence in statin clinical trials: comparing the results. AB - This review of lipid-lowering trials looks at both the primary and secondary prevention of coronary heart disease. It also introduces evidence pointing toward the emerging importance of nonlipid risk factors in the pathogenesis of atherosclerosis. This review is designed to enhance the primary care physician's working knowledge of the evidence supporting the importance of LDL-C reduction and the central role of statin therapy in the reduction of the risk of coronary heart disease, as well as provide a greater understanding of emerging risk factors. PMID- 12884939 TI - Lipid and nonlipid benefits of statins. AB - The Third Report of the National Cholesterol Education Program (NCEP) Expert Panel on Detection, Evaluation, and Treatment of High Blood Cholesterol in Adults (ATP III) provides an evidence-based approach to the diagnosis of dyslipidemia and the management of low-density lipoprotein cholesterol (LDL-C). This article provides a brief overview of the ATP III therapeutic approach (therapeutic lifestyle changes alone or in combination with a pharmacologic agent) in which the patient's level of risk for coronary heart disease guides the intensity of intervention to lower LDL-C concentrations. Because statins have been found to effectively help patients reach ATP III target LDL-C levels to reduce their risk of coronary events, they tend to be the treatment of choice when initial therapy with therapeutic lifestyle changes alone fails to achieve the target level. Discussion includes a summary of the beneficial properties of statins beyond lipid lowering. PMID- 12884940 TI - Clinical information affects electrocardiogram interpretation and can influence patient entry type. PMID- 12884941 TI - Osteopathic graduate medical education opportunities abound in south Florida. PMID- 12884942 TI - Osteopathic graduate medical education opportunities abound in south Florida. PMID- 12884943 TI - Do osteopathic physicians differ in patient interaction from allopathic physicians? An empirically derived approach. AB - Colleges of osteopathic medicine teach osteopathic principles, which provide a different approach to and interaction with patients than principles taught in allopathic medical schools. The authors examined whether osteopathic primary care physicians' interactions with patients reflect the principles of osteopathic medicine when compared with allopathic physicians' interactions. The principles of osteopathic medicine were adapted to elements that could be measured from an audio recording. This 26-item index was refined with two focus groups of practicing osteopathic physicians. Fifty-four patient visits to 11 osteopathic and 7 allopathic primary care physicians in Maine for screening physicals, headache, low back pain, and hypertension were recorded on audiotape and were dual-abstracted. When the 26-item index of osteopathic principles was summed, the osteopathic physicians had consistently higher scores (11 vs. 6.9; P = .01) than allopathic physicians, and visit length was similar (22 minutes vs. 20 minutes, respectively). Twenty-three of the 26 items were used more commonly by the osteopathic physicians. Osteopathic physicians were more likely than allopathic physicians to use patients' first names; explain etiologic factors to patients; and discuss social, family, and emotional impact of illnesses. In this study, osteopathic physicians were easily distinguishable from allopathic physicians by their verbal interactions with patients. Future studies should replicate this finding as well as determine whether it correlates with patient outcomes and satisfaction. PMID- 12884944 TI - Treating patients in primary care: the impact of mood, behavior, and thought disturbances. AB - Patients with symptoms of mood, behavior, and thought disturbances are regularly treated in the primary care setting. More often than not, the disorders associated with these symptoms are overlooked or misdiagnosed by physicians, in part because these patients present symptomatic complaints that are seemingly unrelated to the underlying disorder. Recognition of comorbid psychiatric symptoms allows physicians to treat the whole person more effectively. Furthermore, patients and their caregivers benefit greatly from the early intervention and treatment that is frequently provided in the primary care setting. With the appropriate training so that they may readily recognize these symptoms, osteopathic physicians can help prevent the further progression of--or potential unfavorable outcomes from--otherwise untreated or inadequately treated illnesses. PMID- 12884945 TI - Relationship between academic achievement and student performance on the Comprehensive Osteopathic Medical Licensing Examination-USA level 2. AB - The Comprehensive Osteopathic Medical Licensing Examination-USA (COMLEX-USA) is a three-part examination used for licensing osteopathic physicians. The relationship between performance on this examination and prior student academic performance has been unclear. This study explores the relationship between COMLEX USA level 2 scores and student academic performance at Oklahoma State University College of Osteopathic Medicine (OSU-COM). All first-time examination candidates from OSU-COM had a formal, weeklong board review in March 2000, and all passed the examination. Predictions about COMLEX-USA scores generated by the academic dean's office at OSU-COM underestimated student examination performance; results suggest a significant correlation between level 2 performance with (1) level 1 performance (.751); (2) grade point average (GPA) in basic science (.659); (3) total GPA (.672); and (4) Medical College Admission Test (MCAT) scores (.406). The correlation of level 2 scores to clinical GPA (.269) was lower but still significant. Results of this study suggest that performance on COMLEX-USA level 2 is strongly correlated with prior student academic performance in this population. PMID- 12884946 TI - Predictive validity of osteopathic medical licensing examinations for osteopathic medical knowledge measured by graduate written examinations. AB - The purpose of this study was to evaluate the predictive validity of osteopathic medical licensing examinations for osteopathic medical knowledge measured by graduate written medical examinations. Performances on the three osteopathic initial licensing examinations, the three osteopathic internal medicine in service examinations, and the osteopathic internal medicine board certification examinations were analyzed for a cohort of the most recent osteopathic internal medicine board certification examination candidates (N = 82). Multiple regressions were performed for the predictive value of licensing examination scores for the late examination scores. Logistic regressions were used for the prediction of pass/fail status on the licensing examinations for that on the board certification. A longitudinal performance profile was constructed to assess the rank changes in decile on the examinations at different times. All correlation coefficients between the licensing examinations and other examinations were significant and higher than .70. The licensing examinations together predicted at least 60% of the variance of any of the other examination scores. The pass/fail status on the licensing examinations predicted 89% of the pass/fail status on the certification examination. Decile ranks since the first licensing examination were consistent and stable over at least 5 years. The osteopathic licensing examinations had high predictive validity for the late written osteopathic internal medicine examinations. Generalization of the findings to other disciplines needs a caution, as a specialty bias may exist for these types of studies. PMID- 12884947 TI - Neophaeosphaeria and Phaeosphaeriopsis, segregates of Paraphaeosphaeria. AB - Two new genera, Neophaeosphaeria and Phaeosphaeriopsis, are described to accommodate species of Paraphaeosphaeria that are not congeneric based on morphological characters and results of 18S rDNA sequence analyses. Paraphaeosphaeria s. str. is restricted to species with two-septate ascospores and anamorphs that produce non-septate, smooth, pale brown conidia enteroblastically from phialides which have some periclinal thickening. Species in Neophaeosphaeria have 3-4-septate ascospores and anamorphs that produce ovoid to ellipsoid, non-septate, brown, verrucose or punctate conidia from percurrently proliferating conidiogenous cells. Paraphaeosphaeria barrii, P. conglomerata, P. filamentosa and P. quadriseptata are transferred to Neophaeosphaeria. At present all species in Neophaeosphaeria occur on Yucca (Agavaceae). Phaeosphaeriopsis is described for species that produce 4-5-septate ascospores. Known anamorphs produce cylindrical, 0-3-septate, brown, punctate conidia from percurrently proliferating conidiogenous cells or bacillar conidia from simple phialides. P. agavensis, P. glauco-punctata, P. nolinae and P. obtusispora are transferred to Phaeosphaeriopsis. P. amblyspora is described as a new species. PMID- 12884948 TI - Marchandiobasidium aurantiacum gen. sp. nov., the teleomorph of Marchandiomyces aurantiacus (Basidiomycota, ceratobasidiales). AB - The name Marchandiobasidium aurantiacum gen. sp. nov. is introduced for the teleomorph of Marchandiomyces aurantiacus. Dolipore septa and septal pore caps of the closely related Marchandiomyces corallinus are typical of the Ceratobasidiales, and the basidiomatal characters of Marchandiobasidium aurantiacum are reminiscent of those of the monotypic genus Waitea. Morphological, ultrastructural and molecular data suggest that Marchandiobasidium should not be included in Waitea, but should be treated as a distinct genus. PMID- 12884949 TI - Species specific identification of the Neofabraea pathogen complex associated with pome fruits using PCR and multiplex DNA amplification. AB - Five species of pathogenic fungi belong to Neofabraea. One of these, N. krawtzewii (syn. N. populi), is responsible for bark lesions on poplar (Populus) trees. The other four species cause post-harvest bull's eye rot of pome fruits, and at least two of these also cause bark cankers on pome fruit trees. Morphological variation among these species is slight, and overlap in geographic range sometimes occurs. As a consequence, identification based on conventional criteria can be tenuous. PCR primers with putative species specificity were developed following genetic analysis of the beta-tubulin gene for isolates of each of the five species of Neofabraea. PCR conditions required to achieve specificity of the primer sets were determined, and a multiplex PCR protocol was developed to optimize their diagnostic utility on apple fruits. A protocol with higher annealing temperatures in the initial PCR cycles followed by lower temperatures in later cycles gave complete species-specificity when the primer sets were used individually and in multiplex, resulting in successful detection of the pathogens from axenic culture and infected apple fruits. PMID- 12884950 TI - A molecular phylogeny of Pythium insidiosum. AB - Sequence analysis of the ribosomal DNA internal transcribed spacers (ITS) was used to establish phylogenetic relationships among 23 isolates of Pythium insidiosum, the etiological agent of pythiosis in mammals. The isolates were divided into three distinct clades that exhibited significant geographic isolation. Clade I consisted of isolates from North, Central, and South America, while clade II contained isolates from Asia and Australia. Also present in clade II was an isolate from a patient in the USA, but the origin of the infection may have been in the Middle East. Clade III was comprised of isolates from Thailand and the USA. All 23 P. insidiosum isolates were more closely related to each other than to any other Pythium species in this study. Additionally, all Pythium isolates formed a clade separate from both outgroup species, Phytophthora megasperma and Lagenidium giganteum. The ITS sequence results tend to support the existence of geographic variants or cryptic speciation within P. insidiosum. The sequence information obtained also provides an abundance of data for applications in the diagnosis of pythiosis and identification of P. insidiosum from clinical samples. PMID- 12884951 TI - Molecular genetic variability of Australian isolates of five cereal rust pathogens. AB - Rust fungi cause economically important diseases of cereals, and their ability to rapidly evolve new virulent races has hindered attempts to control them by genetic resistance. PCR-based molecular tools may assist in understanding the genetic structure of pathogen populations. The high multiplex DNA fingerprinting techniques, amplified fragment length polymorphisms (AFLP), selectively amplified microsatellites (SAM) and sequence-specific amplification polymorphisms (S-SAP) were assessed for their potential in investigations of the genetic relationships among isolates of the wheat rust pathogens, Puccinia graminis f. sp. tritici (Pgt), Puccinia triticina (Pt), and P. striiformis f. sp. tritici (Pst), the oat stem rust pathogen P. graminis f. sp. avenae (Pga), and a putative new P. striiformis special form tentatively designated Barley grass yellow rust (Bgyr). Marker information content, as indicated by the number of species-specific fragments, polymorphic fragments among pathotypes, percentage of polymorphic loci, and the marker index, was highest for the SAM assay, followed by the AFLP and S-SAP assays. UPGMA analysis revealed that all marker types efficiently discriminated the five different taxa and Mantel tests revealed significant correlations between the marker types. Within pathogen groups, the marker types differed in the amount of variation detected among isolates; however, the major differences were consistent and polymorphism was generally low. This was reflected by the AMOVA analysis that significantly partitioned 90% of the genetic variation between taxa. Of the three marker types, SAMS were the most informative, and have the potential for the development of locus-specific microsatellites. PMID- 12884952 TI - Multiple gene genealogies and microsatellite markers reflect relationships between morphotypes of Sphaeropsis sapinea and distinguish a new species of Diplodia. AB - Sphaeropsis sapinea is an opportunistic pathogen causing serious damage to conifers, pre-disposed by adverse environmental conditions or mechanical damage. Three different morphological forms of the fungus have been described and are commonly referred to as the A, B and C morphotypes. Isolates of the different morphotypes have also been separated based on differences in pathogenicity and molecular characteristics. These differences, however, overlap and have not been considered sufficiently robust to justify the description of separate taxa. The aim of this study was to consider relationships between isolates representing different S. sapinea morphotypes, using multiple gene genealogies inferred from partial sequences of six protein-coding genes and six microsatellite loci. Genealogies generated for the protein-coding genes and microsatellite loci were not congruent but both consistently grouped isolates representing the A and C morphotypes in separate but closely related clades. In contrast, isolates of the B morphotype grouped together in a clade that was equally different to the A and C morphotypes as it was to the clade encompassing isolates of Botryosphaeria obtusa. These results provide strong evidence to show that the B morphotype isolates are distantly related to S. sapinea and represent a discrete taxon, which we describe here as Diplodia scrobiculata sp. nov. PMID- 12884953 TI - Genetic variation in Corynespora cassiicola: a possible relationship between host origin and virulence. AB - Genetic variation of 42 isolates of Corynespora cassiicola, a destructive fungal pathogen of many economically important crop plants including rubber, was investigated using RAPD-PCR analysis. Five genetic groups were identified using RAPD-PCR profiles generated by eight random primers. Results indicate that there is a significant genetic variation among C. cassiicola isolates collected from different host plants. These results should facilitate the development of rubber clones with enhanced resistance against all genetic groups of C. cassiicola. PMID- 12884954 TI - Telomere fingerprinting for assessing chromosome number, isolate typing and recombination in the entomopathogen Beauveria bassiana. AB - Beauveria bassiana is a popular biocontrol agent used as 'green' pesticide in crop insect pest management. Chromosome number has been variously reported as five, six, seven and eight in this species. The range of chromosome number and the minimum chromosome number in this economically important fungus were assessed through telomere fingerprint analysis of a sample of 17 isolates from different and similar hosts and distant and same geographic origin. Genomic DNA digested with EcoRI, which has no cutting site in the telomere repeat sequence arrays was probed with a radioisotope-labelled (5'-TTAGGG-3')s oligonucleotide. The probe hybridised regions appeared as discrete bands--each representing a telomere. The number of bands in each lane was counted and halved to arrive at the chromosome number of that isolate. The chromosome number varied from 5 to 10 in the different isolates. The telomere probe hybridised bands were also scored for presence or absence in a 0-1 matrix and a dendrogram based on similarities between the isolates was constructed using the NTSYS-pc ver. 2.02i software. The isolates showed very little similarity; the overall similarity was 14%. Only two isolates which were of diverse host and geographic origin showed 100% similarity. Isolates from the same epizootic that showed 43% similarity in their telomere fingerprints had 96 % similarity in their RAPD (Random amplified polymorphic DNA) fingerprints with 10 primers. The genetic distances computed from any one DNA fingerprinting method thus do not reflect the true genetic similarities of the isolates. The frequency distribution pattern of the pair-wise similarities computed from telomere fingerprints hinted at the occurrence of recombination in this fungus. Telomere fingerprinting proved very useful in typing isolates since each of them was found to have a unique fingerprint. Isolates with the same chromosome number neither showed a distinct morphology or virulence character nor a close similarity in telomere or RAPD fingerprints to merit their subgrouping into a taxonomically relevant or practically useful unit. PMID- 12884955 TI - Cytotoxicity against insect cells of entomopathogenic fungi of the genera Hypocrella (anamorph Aschersonia): possible agents for biological control. AB - Extracts from entomopathogenic fungi of the genus Hypocrella (7 species) and its anamorph Aschersonia (11 species) were screened for cytotoxicity to Sf9 and C6/36 insect cells and L929, BHK(21)C13 and HepG2 mammalian cells. Cytotoxic extracts to insect cells (ID50's < or = 10 microg ml(-1)) but not to mammalian cells (ID50's > or = 10 microg ml(-1)) conformed to the criteria of the project and were considered 'lead' extracts for further investigation. 'Leads' were found in two of the Hypocrella species: H. discoidea, and H. tamurai and in three of the Aschersonia species: A. samoensis, A. badia, and A. tamurai. Bioassay-guided fractionation of the cell extract of the fungus A. samoensis BCC 1393 led to the identification of two known anthraquinone dimers, (+)rugulosin (1) and skyrin (2) which showed selective toxicity towards insect cells. (+)Rugulosin (1) and skyrin (2) exhibited strong cytotoxic activity against the insect cell line Sf9 with respective ID50 values of 1.2 and 9.6 microg ml(-1), but showed weak activity toward mammalian cells. This first report of (+)rugulosin (1) and skyrin (2) in A. samoensis is confirmed and demonstrated in another four strains of A. samoensis isolated in Thailand. The preferential cytotoxicity against Sf9 insect cells gives evidence that these insect-pathogenic fungi of the Hypocrella/Aschersonia group might be useful as an agent for pest control. PMID- 12884956 TI - Population dynamics of epiphytic mycoparasites of the genera Clonostachys and Fusarium for the biocontrol of black pod (Phytophthora palmivora) and moniliasis (Moniliophthora roreri) on cocoa (Theobroma cacao). AB - Mycoparasites collected from aerial parts of the cocoa plant (Theobroma cacao) have shown great promise in the control of black pod, caused by Phytophthora palmivora, and moniliasis, caused by Moniliophthora roreri. However, the ecology of epiphytic mycoparasites is still poorly understood although it has a direct bearing on applied biocontrol practices, ranging from the identification and isolation of promising biocontrol candidates to formulation needs and required application frequency. One objective of this study was to determine the natural abundance of mycoparasites on cocoa flowers and pods in relation to crop development stage and cultivar. For this purpose, native mycoparasites were detected on cocoa flowers and pods using the precolonised plate baiting technique. Furthermore, the survival of an applied Clonostachys rosea isolate on cocoa pods on shaded and non-shaded trees was compared as well as the recolonisation patterns of surface-sterilised pods by native mycoparasites under these conditions. Clonostachys spp. were the most commonly isolated native mycoparasites, followed by Fusarium spp. No differences in the occurrence of native, epiphytic mycoparasites were observed between the three main cocoa cultivars, 'Criollo', 'Forastero' and 'Trinitario', nor between clones within these groups. Thus, a single biocontrol inoculum can be suitable for application to cultivar mixtures of cocoa commonly grown together in a field. Different susceptibility classes of segregating F1 populations of hybrids with resistance against M. roreri and P. palmivora supported similar population levels and taxonomic assemblages of mycoparasites. Therefore, we reject the hypothesis that these antagonists mediate resistance. Mycoparasite abundance and genetic disease resistance to black pod and moniliasis are independent phenomena and should lead to additive effects if employed simultaneously in an integrated disease management programme. The survival of applied C. rosea was not affected by the shading regime or any other meteorological parameter measured. On the other hand, recolonisation of surface-sterilised cocoa pods by most native mycoparasites was faster in the shade. Only Trichoderma spp. colonised pods exposed to direct sunlight faster than shaded ones. The implications for the design of biocontrol inocula and formulation technology are discussed. PMID- 12884957 TI - Polyporus s. str. in southern South America: isoenzyme analysis. AB - Forty-two dikaryotic and 42 monokaryotic isolates, and 34 pairings were examined by horizontal polyacrylamide gel electrophoresis (PAGE) for six enzymatic activities, viz. EST, 6PGD, IDH, MDH, SHDH and SOD. 44 bands were analysed. Numerical analysis of the isoenzymatic patterns was undertaken and compared with those from morphological characters. The analysis of six enzymatic systems showed the existence of four monomorphic systems (IDH, MDH, SHDH and SOD). The sterease system (EST) appears to be polymorphic in Polyporus ciliatus and in populations of P. tenuiculus from Argentina, being monomorphic in the remaining species studied. The 6PGD system is polymorphic in P. tucumanensis and monomorphic in the other species. Predominance of monomorphic enzymes and a clear distribution of the electromorphs among the species, indicates that isoenzymatic analysis is a good taxonomic tool within Polyporus. The low intraspecific variability allowed the use of interspecific differences to separate species. Numerical analysis showed a good correlation between morphological and molecular characters. In the isoenzymatic phenogram the similarity index is high only among very close species, showing a stressed separation of species. PMID- 12884958 TI - Differentiation of Agaricus species and other homobasidiomycetes based on volatile production patterns using an electronic nose system. AB - Comparisons of the qualitative volatile production patterns between seven species of Agaricus, and between two of Volvariella and Pleurotus and one Coprinus species when grown at 25 degrees C on agar media for 14 d were made. There was good reproducibility between the volatile production patterns of the same species using an electronic nose unit with a 14 conducting sensor polymer array. Principle Component Analysis (PCA) showed that it was possible to discriminate between five of the seven Agaricus species, but that some overlap occurred between the others. Cluster analysis showed that there was also overlap between some species with the tropical collection of A. bitorquis separating out from the others. The volatile production profile of the commercial A. bisporus was close to that of a wild species, A. campestris. A. bisporus could be readily differentiated from other non-Agaricus species. This study demonstrates the potential for using electronic nose systems to rapidly differentiate mycelial cultures of homobasidiomycete mushrooms. PMID- 12884959 TI - Long-term viability and infectivity of intraradical forms of Glomus intraradices vesicles encapsulated in alginate beads. AB - Intraradical vesicles of Glomus intraradices were isolated, entrapped in alginate beads, and stored at 4 degrees C for periods from 2-74 months. The beads were used to inoculate leeks grown under standard conditions for 6 wk, then development of root colonization by G. intraradices was recorded. Colonization of leeks was high (mean > 50% in length) and did not vary markedly until five years of storage. After six years of storage, the inoculum proved infective and viable. PMID- 12884960 TI - Effect of intraspecific competition by Aspergillus flavus on aflatoxin formation in suspended disc culture. AB - The ability of two non-aflatoxin producing strains of Aspergillus flavus to interfere with aflatoxin production by a toxigenic A. flavus strain was examined using a replacement series with suspended disc culture method. Individual glass fiber discs, affixed to a pin suspended from the caps of scintillation vials, were inoculated with medium containing A. flavus conidial mixtures in different proportions (aflatoxin producer:non-producer = 100:0, 80:20, 60:40, 20:80 and 0:100 by vol) at a constant total density (1 x 10(5) spores ml(-1)). Reductions in the total conidial density of these strains when grown alone, had little effect on fungal growth (mycelium dry weight) or aflatoxin production. Significant (P < 0.0001) reductions in aflatoxin B1 were recorded when non toxigenic strains represented any proportion of the inoculum mixture. Aflatoxin yield values were less than (P < 0.0001) expected from the input ratios for toxigenic vs. non-toxigenic conidial inoculum within the replacement series. Aflatoxin yields were also reduced (P < 0.001), with a corresponding increase in fungal growth (P < 0.001), when conidia from aflatoxin producing strains were mixed in equal proportions. This suggests that the substantial inhibition of aflatoxin yield for inoculum mixtures results from the failure of spore germlings to establish a cooperative mycelial network. PMID- 12884961 TI - Beta-carotene production and sclerotial differentiation in Sclerotinia minor. AB - Sclerotinia minor accumulates beta-carotene at levels dependent upon oxidative growth conditions and differentiation. Beta-carotene accumulation is 2.5-fold higher in differentiated mycelia at high than at low oxidative stress, and approx. 3-fold higher in differentiated than in undifferentiated mycelia. It is proposed that beta-carotene may be produced by the fungus to counteract oxidative stress that develops during growth. This is shown by the finding that exogenous beta-carotene at growth non-inhibiting concentrations causes a concentration dependent reduction of oxidative stress (lipid and protein peroxidation) and sclerotial differentiation in this fungus. The data of this study support our hypothesis that sclerotial differentiation in phytopathogenic fungi may be induced by oxidative stress. PMID- 12884962 TI - Assemblage structure, species richness, abundance, and distribution of fungal fruit bodies in a seven year plot-based survey near Vienna. AB - Almost 900 species were encountered during a seven year survey of fungal fruit bodies in 13 forests and grasslands plots measuring about 1 ha. The data were comparable to those from a Swiss forest plot monitored for 21 years. Species richness and abundance were almost linearly correlated on a log/log scale. Abundance and yearly frequency of species were tightly correlated. The geographic range of species may also be related to these two parameters. Rare species scored low on abundance, yearly frequency and geographic range. About half of the species were rare, and occurred in only one out of seven years. The two major functional groups of saprotrophic and mycorrhizal species behaved similarly over the years, as in the Swiss study. Annual variation was high for species richness alone and for richness combined with abundances. The log transformed species richness of plots correlated well with parameters that take the abundances or yearly frequencies of species into account. If other mushroom assemblages show similar regularities, it may be possible to omit abundance counts in future surveys. The species composition of the plots varied strongly over the years. Many species did not reach their maximum abundance in the richest year. Species showed their maximum abundances in different plots in the same year more often than expected by chance. Thus the presence and abundance of species depended on factor(s) other than the general productivity of a year, and years showed (a) different aspect(s) than productivity alone. Long-term surveys are important for understanding the structure of mushroom assemblages and their biodiversity. PMID- 12884963 TI - Altered conditioned fear behavior in glutamate decarboxylase 65 null mutant mice. AB - We investigated the involvement of the 65 kDa isoform of glutamic acid decarboxylase (GAD65) and GAD65-mediated gamma-aminobutyric acid (GABA) synthesis in the formation and expression of Pavlovian fear memory. To this end, behavioral, endocrine and autonomic parameters were examined during conditioned fear retrieval of mice with targeted ablation of the GAD65 gene (GAD65-/- mice). These mutant mice were found to display specific fear behavior (freezing, escape), as well as autonomic (increased defecation) and endocrine activation (increased plasma corticosterone) during fear memory retrieval. However, freezing was reduced and flight and escape behavior were increased in GAD65-/- mice compared to their wild type and heterozygous littermates, while corticosterone levels and defecation rates did not differ between genotypes. Active defensive behavior of GAD65-/- mice was observed during both auditory cued and contextual retrieval of fear memory, as well as immediately after conditioning. These data indicate a selectively altered behavioral fear response in GAD65-/- mice, most likely due to deficits in threat estimation or the elicitation of appropriate conditioned fear behavior, and suggest that GAD65 is a genetic determinant of conditioned fear behavior. GAD65-/- mice provide a valuable tool to further dissect the GABAergic mechanisms involved in fear and anxiety and to model GABA related neurological and psychiatric disorders. PMID- 12884964 TI - A rating scale for wildness and ease of handling laboratory mice: results for 21 inbred strains tested in two laboratories. AB - Rating scales for difficulty in capturing and holding mice were devised that proved to be easy to use and highly sensitive to differences among mouse strains on the A and B priority lists of the Mouse Phenome Project. The simplicity of the scales makes it feasible to rate wildness during behavioral test sessions without adding much to testing time or distracting the technician from the principal task at hand. Overall wildness and placidity ratings obtained by combining capture and hold ratings provide a good impression of the difficulties encountered while working with lab mice in the course of complex experiments. Ratings of 21 inbred strains during the course of 15 behavioral tests in two laboratories demonstrated that the SPRET/Ei, PERA/Ei, CAST/Ei and SWR/J strains were particularly difficult to handle. The NOD/LtJ strain posed no special challenge in the Edmonton laboratory but was very difficult to handle in the Portland lab. The rating scales should be useful for judging the difficulties in working with novel targeted or induced mutations in mice as well as effects of a variety of environmental treatments or drugs. PMID- 12884965 TI - Differential involvement of the mu and kappa opioid receptors in spatial learning. AB - In order to test the role of mu and kappa opioid receptors (Mu opioid receptor (MOR) and Kappa opioid receptor (KOR)) in hippocampal-dependent spatial learning, we analyzed genetically engineered null mutant mice missing the functional MOR or KOR gene. Compared to wild-type mice, the homozygous MOR null mutants exhibited an impairment in the ultimate level of spatial learning as shown in two distinct tasks, the 8-arm radial-maze and the Morris water-maze. Control behaviors were normal. The learning impairment could be associated with the impairment we found in the maintenance of long-term potentiation in mossy fibers in CA3. In comparison, there was no impairment in spatial learning in our KOR mutants or in mossy fibers (mf) in CA3 region long-term potentiation (LTP). Our work suggests that the MOR may play a positive role in learning and memory by increasing LTP in CA3 neurons. PMID- 12884966 TI - Prepulse inhibition of acoustic startle in aromatase knock-out mice: effects of age and gender. AB - Estrogen has been suggested to play a neuromodulatory and neuroprotective role on the brain dopamine system. We used aromatase knockout (ArKO) mice that lack a functional aromatase enzyme and are unable to convert testosterone into estrogen, and assessed prepulse inhibition of acoustic startle, locomotor hyperactivity to amphetamine treatment and rotarod performance. Mice were tested at either 1 month, 4-5 months or 12-18 months of age. In male, but not female ArKO mice, there was an age-related reduction of prepulse inhibition. The 12-18 months old male ArKO mice also showed significantly greater amphetamine-induced hyperactivity. Mice heterozygous for the mutation showed no deficits or were in between wildtype mice and ArKO mice. We postulate that these data indicate a neuroprotective role of estrogen, particularly in male mice, on ageing of brain mechanisms involved in pre-pulse inhibition and locomotor activity regulation. It is likely that these brain mechanisms are or include dopaminergic activity. PMID- 12884967 TI - Hippocampal infection with HSV-1-derived vectors expressing an NMDAR1 antisense modifies behavior. AB - Herpes simplex virus-derived amplicon vectors simultaneously expressing the open reading frame encoding NR1 subunit of the NMDA receptor, either in sense or antisense orientation, as well as the open reading frame encoding the green fluorescent protein (GFP), as distinct transcription units, were constructed. Vector expression in cells was demonstrated by GFP-fluorescence, immunofluorescence, Western blots and RT-PCR. The vectors were inoculated into the dorsal hippocampus of adult male rats, which were then trained for habituation to an open field and for inhibitory avoidance to a foot-shock. Those animals injected with vectors expressing NR1 protein showed habituation to a new environment, and achieved the criteria for a step-down inhibitory avoidance to a foot-shock. In contrast, animals injected with vectors carrying the NR1 open reading frame in antisense position, showed neither habituation nor appropriate performance in the inhibitory avoidance task. There was no evidence for motor impairment or motivational disturbance, since all the animals exhibit similar behavior and performance in the training sessions. Hence, the impaired performance might be due to either amnesia or disability to record events. Transgene expression in brain, as revealed by GFP fluorescence, was mainly observed in pyramidal cells of CA1, but also in CA3. Therefore, our results strongly support the participation of hippocampal NR1 subunit in habituation to a new environment, but also in recording events for the inhibitory avoidance task. Hence, amplicon vectors appear to be useful tools to modify endogenous gene expression at a defined period, in restricted brain regions, and should allow investigating in vivo functions of genes. PMID- 12884968 TI - Congenic C57BL/6 mu opiate receptor (MOR) knockout mice: baseline and opiate effects. AB - Homozygous mu-opioid receptor (MOR) knockout (KO) mice developed on a chimeric C57B6/129SV background lack morphine-induced antinociception, locomotion and reward. Therefore it appears that MOR largely mediates these morphine actions. However, one factor that could affect the extent of knockout deficits in morphine induced behavior is the genetic background against which the gene deletion is expressed. To examine the effect of genetic background chimeric C57B6/129SV MOR knockout mice from the 15th generation of those developed in our laboratory were backcrossed for 10 successive generations with C57BL/6 mice, a strain which is more sensitive to many of the properties of morphine, to produce congenic MOR (con-MOR) KO mice. Heterozygote conMOR KO mice display attenuated morphine locomotion and reduced morphine analgesia compared to wild-type mice. Homozygote con-MOR KO mice display baseline hyperalgesia, no morphine place preference, no morphine analgesia and no morphine locomotion. These results are not qualitatively different from those observed in the MOR KO strain with a chimeric C57B6/129SV background, and suggest that although the strain has separate influences on these functions, it does not substantially interact with deletion of the mu opiate receptor gene. PMID- 12884969 TI - Transgenic and knock-out mouse pups: the growing need for behavioral analysis. AB - Few laboratories working with transgenic and knockout mice analyze the neurobehavioral consequences of genetic manipulation in early ontogeny. However, the study of behavioral endpoints during the early postnatal period in genetically modified mice is important not only to assess possible developmental abnormalities, but also to better understand and disentangle the effects of genetic manipulations in adulthood. We propose that the assessment of neurobehavioral development represents an appropriate strategy to identify possible compensatory and/or unexpected effects. Nowadays, a large number of experimental protocols that take into account the practical constraints imposed by the peculiar physiological and behavioral responses of an immature subject are available to assess the neurobehavioral profile of developing mice. While this knowledge should be applied to the field of transgenic and knock-out mice in general, it should be recommended, in particular, for the study of mouse models of neurodevelopmental disorders. PMID- 12884970 TI - Does my mouse have Alzheimer's disease? AB - Small animal models that manifest many of the characteristic neuropathological and behavioral features of Alzheimer's disease (AD) have been developed and have proven of great value for studying the pathogenesis of this disorder at the molecular, cellular and behavioral levels. The great progress made in our understanding of the genetic factors that either cause or contribute to the risk of developing AD has prompted many laboratories to create transgenic (tg) mice that overexpress specific genes which cause familial forms of the disease. Several of these tg mice display neuropathological and behavioral features of AD including amyloid beta-peptide (A beta) and amyloid deposits, neuritic plaques, gliosis, synaptic alterations and signs of neurodegeneration as well as memory impairment. Despite these similarities, important differences in neuropathology and behavior between these tg mouse models and AD have also been observed, and to date no perfect animal model has emerged. Moreover, ascertaining which elements of the neuropathological and behavioral phenotype of these various strains of tg mice are relevant to that observed in AD continues to be a challenge. Here we provide a critical review of the AD-like neuropathology and behavioral phenotypes of several well-known and utilized tg mice that express human APP transgenes. PMID- 12884971 TI - Strain and sex differences in repeated ethanol treatment-induced motor activity in quasi-congenic mice. AB - The B6.C quasi-congenic Recombinant QTL Introgression (RQI) strains of the b4i5 series have similar genetic background, but differ in about 5% of their genome from the C57BL/6ByJ (B6) background strain because they carry short chromosome segments introgressed from the BALB/cJ (C) donor strain. These RQI strains were derived from mouse lines selectively bred for high activity of mesencephalic tyrosine hydroxylase (TH/MES), therefore genetic variation in dopamine system related behaviours, such as ethanol-induced motor activity, can be expected. Males and females of 17 RQI and two progenitor strains were tested for initial motor activity for 15 min after a habituating injection of saline, which was followed by an i.p. injection of saline or ethanol (2 g/kg) and an additional test of motor activity for 30 min. This procedure was repeated during 4 subsequent days. In all strains, the first-day ethanol treatment showed an inhibitory effect. With repetition of the treatment the inhibitory effect decreased, and a stimulatory effect could be observed with significant strain- and sex-dependent variation. Females exhibited higher activity in the saline group than males, and reached an equilibrium of inhibition and stimulation sooner than males with repetition of the ethanol treatment. The highest (> 25-fold) difference in activity after repeated ethanol treatment was detected between females of the two strains B6.Cb4i5-Alpha4/Vad and B6.Cb4i5-Beta13/Vad. These results firstly suggest that females are more sensitive to repeated ethanol exposure than males, secondly they support the observations that ethanol has both inhibitory and stimulatory effects on motor activity, which are affected by sex, genotype, and repetition of treatment, and thirdly offer new quasi-congenic animal models with highly different responses to ethanol allowing one to more quickly move to gene detection. PMID- 12884972 TI - Home-cage activity in heterogeneous stock (HS) mice as a model of baseline activity. AB - Behavioral genetic work in humans indicates that clinical hyperactivity is best viewed as the extreme end of activity levels in the population. However, current animal models of hyperactivity are not studied as quantitative traits as they are either knockout models or inbred strains. Furthermore, these animal models generally demonstrate elevated locomotion in novel environments, but not in their home-cages. This is the opposite of the symptoms seen in the human condition where childhood hyperactivity is generally more pronounced in constant, unstimulating situations. In this study we filmed an outbred population of 44 heterogeneous stock (HS) mice under red light during their active phase, to assess the reliability of individual differences in home-cage behavior and extract an index of home-cage activity (HCA) level. We then compared this measure to locomotor behavior in a novel environment--the open-field. Reliable individual differences in home-cage behaviors such as running, swinging on bars, and burrowing were found, and principal component factor analysis yielded a general activity factor, which accounted for 32% of the variance and correlated 0.90 with a subjective impression of activity level. The correlation between HCA and locomotor activity in the open-field was 0.23, which was non-significant. However, the association with HCA level appeared to increase over the five minutes of the open-field, presumably as the mice habituated. Furthermore, although mice displaying particularly high and low HCA were indistinguishable early in the open-field task, they became significantly differentiated over time. We conclude that home-cage behaviors and the open-field, after habituation, display good face and construct validity, and may provide a good model of baseline activity for quantitative trait loci (QTL) discovery and functional genomics in the HS mice. PMID- 12884973 TI - Mice overexpressing CRH show reduced responsiveness in plasma corticosterone after a5-HT1A receptor challenge. AB - Corticotropin-releasing hormone (CRH) overproduction and serotonergic dysfunction have both been implicated in a range of psychiatric disorders, such as anxiety and depression, and several studies have shown interactions between these two neurotransmitter systems. In this study, we investigated the effects of CRH challenge on hypothalamo-pituitary-adrenal (HPA) axis activity in female transgenic mice overproducing CRH. Furthermore, the effects of mild stress on HPA axis activity and body temperature were investigated in these mice. Pre- and post synaptic 5-HT1A receptor function were studied by monitoring body temperature and plasma corticosterone levels after challenge with the 5-HT1A receptor agonist 8 hydroxy-2-(di-n-propyl-amino)-tetralin (8-OH-DPAT). Hypothermia in response to 8 OH-DPAT treatment did not differ between transgenic and wild type mice, indicating unaltered somatodendritic 5-HT1A autoreceptor function in mice overproducing CRH. In wild type mice 8-OH-DPAT increased plasma corticosterone levels, but not in transgenic animals. CRH injection, however, increased corticosterone levels in both groups. These data suggest desensitization of post synaptic, but not pre-synaptic, 5-HT1A receptors in mice overproducing CRH. These findings resemble those seen in depressed patients following 5-HT1A challenge, which is in accord with the hypothesized role of CRH in the pathogenesis of depression. PMID- 12884974 TI - Difference in anxiety and sensitization of the acoustic startle response between the two inbred mouse strains BALB/cAN and DBA/2N. AB - The inbred mouse strain BALB has been proposed to be an animal model for pathological anxiety. BALB exhibits a stronger acoustic startle response (ASR) than the 'less emotional' inbred strain DBA. Four experiments were conducted to determine whether this strong ASR is due to a higher anxiety level and/or to greater sensitization in BALB than in DBA, with the following results: (1) The ASR to the very first startle stimulus was found to be much stronger in BALB than in DBA, and freezing behavior evoked by startle stimuli was more pronounced in BALB than in DBA. These findings indicate a higher level of anxiety in this strain. (2) ASR amplitudes of BALB initially rose much higher during consecutive startle stimuli and remained at a high level much longer than in DBA. Thereafter, ASR amplitude dropped more slowly and to a lesser degree than in DBA. Startle amplitudes decreased similarly in both strains (strong exponential decrease) only when a low sound pressure level (SPL) was used which elicited approximately the same low ASR in both strains. These results can only be explained by increased sensitization in BALB. (3) The slope of the i/o-function, which represents the relation between sensory input and motor output, was steeper in BALB than in DBA. As it has been shown recently, sensitization increases the slope of the startle i/o-function indicating increased sensitization in BALB. It is discussed, however, whether anxiety also contributes to this effect. (4) Footshocks increased the ASR much less in BALB than in DBA, again showing increased sensitization in BALB. Both a higher level of anxiety and greater sensitization therefore determined the greater strength of the ASR in BALB than in DBA. PMID- 12884975 TI - Screening the human protocadherin 8 (PCDH8) gene in schizophrenia. AB - Abnormalities in synaptic connectivity and plasticity have been implicated in the pathophysiology of schizophrenia. Molecules involved in the development and maintenance of neural circuitry include the recently cloned protocadherins. Human protocadherin 8 (PCDH8) is homologous to 'arcadlin', a molecule shown to play a role in hippocampal synaptic function in the rat. The gene encoding PCDH8 maps to a region on chromosome 13 where linkage to schizophrenia has been reported. In this study, the entire expressed sequence of the PCDH8 gene and over 800 bp of the 5' flanking region were screened for polymorphisms in 30 DSM-IV schizophrenia individuals using Denaturing High Performance Liquid Chromatography (DHPLC). A total of nine single nucleotide polymorphisms were identified, including three in the first exon that are predicted to change the amino acid sequence. One polymorphism, causing the Trp7Arg change in the putative signal peptide, showed a trend towards excess of the arginine encoding allele in a case-control sample consisting of 520 DSM-IV schizophrenia patients and 535 matched controls from the UK (chi2=3.72, P [1 df]= 0.054). However, this polymorphism did not show preferential transmission to schizophrenic individuals in a separate sample of 203 proband-parent trios from Bulgaria. A second, rare single nucleotide variation, predicting the non-conservative amino acid change Glu39Ala, was found in one schizophrenic individual and their affected sibling but not in a further 352 affected individuals, nor 357 controls. These results suggest that any contribution of PCDH8 polymorphisms to schizophrenia susceptibility is likely to be weak, although the existence of rare variations of stronger effect cannot be excluded. PMID- 12884976 TI - 'My mouse has no phenotype'. PMID- 12884977 TI - Behavioural and physiological characterization of inbred mouse strains: prospects for elucidating the molecular mechanisms of mammalian learning and memory. AB - With the advent of recombinant DNA methodology, it has become possible to dissect the molecular mechanisms of complex traits, including brain function and behaviour. The increasing amount of available information on the genomes of mammalian organisms, including our own, has facilitated this research. The present review focuses on a somewhat neglected area of genetics, one that involves the study of inbred mouse strains. It is argued that the use of inbred mice is complementary to transgenic approaches in the analysis of molecular mechanisms of complex traits. Whereas transgenic technology allows one to manipulate a single gene and investigate the in vivo effects of highly specific, artificially induced mutations, the study of inbred mouse strains should shed light on the roles of naturally occurring allelic variants in brain function and behaviour. Systematic characterization of the behavioural, electrophysiological, neurochemical, and neuroanatomical properties of a large number of inbred strains is required to elucidate mechanisms of mammalian brain function and behaviour. In essence, a 'mouse phenome' project is needed, entailing the construction of databases to investigate possible causal relationships amongst the phenotypical characteristics. This review focuses on electrophysiological and behavioural characterization of mouse strains. Nevertheless, it is emphasized that the full potential of the analysis of inbred mouse strains may be attained if techniques of numerous disciplines, including gene expression profiling, biochemical analysis, and quantitative trait loci (QTL) mapping, to name but a few, are also included. PMID- 12884978 TI - Ethanol-induced activation and rapid development of tolerance may have some underlying genes in common. AB - Ethanol exerts biphasic effects on behavior, stimulant at low doses and depressant at higher doses. In the present study we used two mouse genetic models to investigate the relationships among activating and depressant responses to alcohol. The first model was a panel of nine isogenic genotypes. FAST and SLOW mice, selectively bred for high and low ethanol-induced motor activation, respectively, were used as a second model. We used loss of righting reflex to assess initial sensitivity and acute functional tolerance to a hypnotic dose of ethanol (3 g/kg, 20% v/v). Blood ethanol concentration at the onset of loss of righting reflex was used as an estimate of initial sensitivity, while the difference between concentration values at the recovery and loss of righting represented an acute functional tolerance score. Mean initial sensitivity and acute functional tolerance values of the nine strains were correlated with a previously obtained measure of ethanol-induced locomotor activation. Activation correlated significantly with both initial sensitivity (rg = 0.80; P < 0.05) and acute functional tolerance (rg = 0.77; P < 0.05). Thus, inbred genotypes that were activated more by a low dose of ethanol were also more sensitive to and developed more acute tolerance to a high dose. FAST mice had initial sensitivity values similar to those of SLOW mice, but developed more pronounced tolerance, indicating that ethanol-induced activation and acute functional tolerance may be regulated by some common genetic mechanisms. In summary, these results supported a genetic association between ethanol-induced activation and rapid development of tolerance. PMID- 12884979 TI - Evidence for general cognitive ability (g) in heterogeneous stock mice and an analysis of potential confounds. AB - The heterogeneous stock (HS) is a genetically outbred line of mice established more than 30 years ago from an 8-way cross of C57BL/6, BALB/c, RIII, AKR, DBA/2, I, A/J and C3H inbred mouse strains. The present study compared the performance of 40 HS mice across a battery of diverse cognitive tasks under a variety of motivations. Indices of emotionality were also included in order to assess their influence on performance. All measures of ability loaded positively on an unrotated first principal component that accounted for 31% of the variance, suggesting the presence of a common factor of general cognitive ability (g) underlying all tasks. A first factor derived from anxiety indices correlated nonsignificantly with all cognitive tasks and nonsignificantly with this g factor, supporting the hypothesis that the factor is cognitive rather than temperamental in nature. The factor was also robust in relation to outliers and sex differences, accounting for 28% of the variance after removal of outlier individuals and also after correcting for variance owing to sex differences. A general cognitive ability (g) appears to underlie the performance of HS mice on a battery tapping diverse cognitive demands. PMID- 12884980 TI - Strain differences in activity and emotionality do not account for differences in learning and memory performance between C57BL/6 and DBA/2 mice. AB - This study examined emotionality, activity, learning and memory, as well as the influence of emotionality and activity on learning and memory performance in C57BL/6 and DBA/2 mice using a mouse-test battery. DBA/2 mice performed more poorly than C57BL/6 mice in complex learning tasks such as the water maze and object recognition tasks. In contrast, C57BL/ 6 mice showed attenuated habituation tonovelty in the open field apparatus and poorer performance in the step-down passive avoidance task. The C57BL/6 mice were less exploratory and more anxious than the DBA/ 2 mice. The anxiety score (open arm entries in the elevated plus maze) was significantly correlated with all measures of learning and memory in the object recognition task, and some measures in the passive avoidance and water maze tasks. Analysis of covariance (with open arm entries as a covariate) revealed that some measures on trial 1 of the object recognition task, but not the memory scores on trial 2,were confounded by anxiety. No confounding factors of anxiety were found in the water maze or passive avoidance tasks. Similar results were obtained with the activity scores (line crossing and rearing in the open field). In conclusion, strain differences in activity and anxiety did not account for strain differences in learning and memory performance of C57BL/6 and DBA/2 mice. Nonetheless, the importance of using complete behavioural test batteries should be stressed to ensure that strain differences in learning and memory tasks are not confounded by non-cognitive factors. PMID- 12884981 TI - Marker-assisted selection of a neuro-behavioural trait related to behavioural inhibition in the SHR strain, an animal model of ADHD. AB - The search for the molecular bases of neuro-behavioural traits in Spontaneously Hypertensive Rats (SHR), an animal model of Attention Deficit Hyperactivity Disorder (ADHD), led to the discovery of two quantitative trait loci related to the locomotor activity in the centre of the open field. In the present study, rats from an F2 intercross between the SHR and Lewis strains were selected with markers on the basis of their genotype at these two loci. We obtained a 'high line' in which rats have the alleles increasing the trait, and a 'low line' with the lowering alleles. In activity cages with a dim light, the low line was more active than the high line. The reverse was found in the open field, and the inhibition of locomotor activity in the low line (as compared to the high line) was directly related to the aversiveness of the situation (larger in the centre than in the periphery, and in high light than in low light), and was more intense in males than in females. This inhibition is not attributable to a classical 'anxiety' factor as measured in the elevated plus maze, in which the open arms behaviours were not different between the lines. The high line also showed a deficit in prepulse inhibition of the acoustic startle reflex. The present data show that the two loci previously described in a SHR x Lewis intercross as related to the activity in the centre of the open field are indeed involved in a behavioural inhibition trait. The marker-based selected lines described here are unique tools for the study of the neurobiological bases of this trait and the molecular foundations of its variability of genetic origin. PMID- 12884983 TI - Resuscitative hypothermia after cardiac arrest in adults. PMID- 12884982 TI - Hot topics in behavioral and neural genetics. PMID- 12884984 TI - Risk stratification for adverse outcome in cardiac surgery. AB - Risk-adjusted outcome prediction is mainly important in two separate fields. The first is quality monitoring: measuring actual versus predicted mortality in an institution allows assessment of the clinical surgical and anaesthesia performance while adjusting for the risk profile of the patients. Without risk stratification, surgeons and hospitals treating high-risk patients will appear to have worse results than others. This may prejudice referral patterns, affect the allocation of resources and even discourage the treatment of high-risk patients. The second field is that of informed consent and clinical decision-making. Risk adjusted predicted mortality should form an important part of patient and surgeon decisions on whether or not to proceed with surgery. Clearly, no 'perfect' model can be produced as some aspects of mortality will always be related to risk factors not included in the model (e.g. the quality of the distal coronary artery vessels in coronary artery surgery) or due to chance happenings not related to preoperative patient characteristics (such as surgical error). An individual patient will either survive or die after cardiac surgery. Clearly, no scoring system will predict the specific outcome for every patient. However, risk stratification will inform patients and clinicians of the likely risk of death for a group of patients with a similar risk profile undergoing the proposed operation. This information is useful and should form part of the basis on which the patient and surgeon decide whether to proceed. PMID- 12884985 TI - Multicentre evaluation of in vitro contracture testing with bolus administration of 4-chloro-m-cresol for diagnosis of malignant hyperthermia susceptibility. AB - BACKGROUND AND OBJECTIVE: The in vitro contracture test with halothane and caffeine is the gold standard for the diagnosis of susceptibility to malignant hyperthermia (MH). However, the sensitivity of the in vitro contracture test is between 97 and 99% and its specificity is 78-94% with the consequence that false negative as well as false-positive test results are possible. 4-Chloro-m-cresol is potentially a more specific test drug for the in vitro contracture test than halothane or caffeine. This multicentre study was designed to investigate whether an in vitro contracture test with bolus administration of 4-chloro-m-cresol can improve the accuracy of the diagnosis of susceptibility to MH. METHODS: Three hundred and fifty-two patients from 11 European MH laboratories participated in the study. The patients were first classified as MH susceptible, MH normal or MH equivocal by the in vitro contracture test according to the European MH protocol. Muscle specimens surplus to diagnostic requirements were used in this study (MH susceptible = 103 viable samples; MH equivocal = 51; MH normal = 204). 4-Chloro-m cresol was added to achieve a concentration of 75 micromol L(-1) in the tissue bath. The in vitro effects on contracture development and muscle twitch were observed for 60 min. RESULTS: After bolus administration of 4-chloro-m-cresol, 75 micromol L(-1), 99 of 103 MH-susceptible specimens developed marked muscle contractures. In contrast, only two of 204 MH-normal specimens showed an insignificant contracture development following 4-chloro-m-cresol. From these results, a sensitivity rate of 96.1% and a specificity rate of 99.0% can be calculated for the in vitro contracture test with bolus administration of 4 chloro-m-cresol 75 micromol L(-1). Forty-three patients were diagnosed as MH equivocal, but only specimens from 16 patients developed contractures in response to 4-chloro-m-cresol, indicating susceptibility to MH. CONCLUSIONS: The in vitro contracture test with halothane and caffeine is well standardized in the European and North American test protocols. However, this conventional test method is associated with the risk of false test results. Therefore, an improvement in the diagnosis of MH is needed. Regarding the results from this multicentre study, the use of 4-chloro-m-cresol could increase the reliability of in vitro contracture testing. PMID- 12884986 TI - QTc dispersion as a marker for medical complications after severe subarachnoid haemorrhage. AB - BACKGROUND AND OBJECTIVE: Morbidity from subarachnoid haemorrhage is common and results from complications including myocardial dysfunction and neurogenic pulmonary oedema causing hypotension and hypoxia--both major causes of secondary brain injury. Predicting patients at risk of developing these complications may facilitate early intervention. METHODS: Using QTc dispersion to assess repolarization inhomogeneity, patients who had suffered severe acute subarachnoid haemorrhage were studied in an intensive care unit. Electrocardiograms were recorded within 24 h of ictus. Subsequent development of myocardial dysfunction was defined as a requirement for inotropes, and neurogenic pulmonary oedema as a PaO2 (kPa)/FiO2 ratio < 40. Together they constituted cardiorespiratory compromise. RESULTS: Twenty-seven patients were recruited. QTc dispersion was greater in patients (74.1 ms, SD +/- 26.1) than in controls (48.3 ms, 12.0) P < 0.0001, 95% CI 14.6, 37.0. Thirteen patients developed cardiorespiratory compromise and had greater QTc dispersion (84.5 ms, 26.2) than patients who did not develop cardiorespiratory compromise (64.5 ms, 22.7) P = 0.046, 95% CI 0.3, 39.6. There was no difference in QTc dispersion between patients who did and those who did not develop myocardial dysfunction alone. Similarly, there was no difference in QTc dispersion between patients who did and those who did not develop neurogenic pulmonary oedema alone. CONCLUSIONS: Increased QTc dispersion is associated with the later development of cardiorespiratory compromise in poor grade subarachnoid haemorrhage patients. QTc dispersion may be used as a marker to predict impending clinical deterioration, providing an opportunity for early intervention. PMID- 12884987 TI - Effects of sevoflurane on cerebral blood flow and cerebral metabolic rate of oxygen in human beings: a comparison with isoflurane. AB - BACKGROUND AND OBJECTIVE: Isoflurane is commonly used for neurosurgery but the effects of sevoflurane on human cerebral blood flow and cerebral metabolic rate of oxygen have not been fully evaluated. We therefore assessed the effects of sevoflurane and isoflurane on global cerebral blood flow and cerebral metabolic rate of oxygen in patients without noxious stimuli or neurological disorders. METHODS: General anaesthesia was induced with midazolam (0.2 mg kg(-1)) and fentanyl (5 microg kg(-1)) in 20 ASA I patients undergoing knee joint endoscopic surgery. Epidural anaesthesia was also performed to avoid noxious stimuli during surgery. Cerebral blood flow and cerebral arteriovenous oxygen content difference was measured using the Kety-Schmidt method with 15% nitrous oxide as a tracer before and after administration of either sevoflurane or isoflurane (1.5 minimum alveolar concentration, 60 min) and cerebral metabolic rate of oxygen was then calculated. RESULTS: Sevoflurane and isoflurane both increased cerebral blood flow (17%, P < 0.05; 25%, P < 0.05, respectively) and decreased cerebral metabolic rate of oxygen (26%, P < 0.01; 38%, P < 0.01, respectively). There were no significant differences in cerebral blood flow and cerebral metabolic rate of oxygen between sevoflurane and isoflurane. CONCLUSIONS: Sevoflurane and isoflurane similarly increased cerebral blood flow and decreased cerebral metabolic rate of oxygen in human beings anaesthetized with midazolam and fentanyl. PMID- 12884988 TI - Inhalational induction of anaesthesia with 8% sevoflurane in children: conditions for endotracheal intubation and side-effects. AB - BACKGROUND AND OBJECTIVE: This study was designed to assess the conditions for endotracheal intubation or insertion of a laryngeal mask airway following an inhalational induction using 8% sevoflurane and nitrous oxide without the use of muscle relaxants or opioids. METHODS: There were two groups: 30 children had endotracheal intubation and 30 children had a laryngeal mask airway inserted. Induction of anaesthesia was accomplished using an inspiratory concentration of sevoflurane 8% in a nitrous oxide and oxygen mixture. After an end-expiratory concentration of sevoflurane of at least 4% had been reached, when the pupils were miotic and centred, the trachea was intubated or a laryngeal mask inserted. The time to loss of consciousness and successful airway management was recorded. Jaw relaxation, movements, visibility, and position of the vocal cords and vital parameters were monitored. RESULTS: Jaw relaxation was complete in all children. The vocal cords were completely visible in all patients of the tracheal intubation group, whereas vocal cord relaxation was incomplete in five children. Nevertheless, all children had an atraumatic intubation or insertion of the laryngeal mask without the use of a muscle relaxant. Vital signs were stable in both groups. There were no cases of restlessness and/or postoperative shivering. Four patients in the endotracheal group (13.3%) were nauseous and three (10%) vomited, while two children (6.6%) in the laryngeal mask group experienced nausea and vomiting. CONCLUSIONS: Induction with sevoflurane in nitrous oxide and oxygen leads to fast loss of consciousness and provides ideal conditions for managing the airway without supplemental opioids or muscle relaxants. Furthermore, sevoflurane using this technique was very well tolerated, indicated by high haemodynamic stability and a reduced rate of postoperative restlessness, shivering, nausea and vomiting. PMID- 12884989 TI - Nitrous oxide concentrations in maternal and fetal blood during caesarean section. AB - BACKGROUND AND OBJECTIVE: There are little data on nitrous oxide (N2O) concentrations in neonatal blood at delivery. We investigated the effects of the time elapsing between the induction of anaesthesia and delivery (the I-D interval) on umbilical blood N2O concentrations. METHODS: Maternal and neonatal blood N2O concentrations were measured in 27 patients undergoing Caesarean section under N2O 67% anaesthesia. The duration of N2O administration (range 2-50 min) was arbitrarily divided into three groups (each n = 9): short (2-9 min), medium (9.1-14 min) and long duration (14.1-50 min). RESULTS: Compared with a rapid increase in the maternal arterial N2O concentration (48.9 +/- 4.7%), the umbilical venous N2O concentration (17.9 +/- 8.3%) rose slowly in the short duration group, whereas the N2O concentrations became more similar (61.6 +/- 4.3 and 43.2 +/- 10.0%, respectively) in the long duration group. The ratio of umbilical vein to maternal artery N2O concentrations correlated with the duration of N2O anaesthesia (r = 0.739), resulting in ratios of 0.37 +/- 0.18, 0.61 +/- 012 and 0.70 +/- 0.13 for the short, medium and long duration groups, respectively. The Apgar score at 1 min correlated inversely with the duration of anaesthesia and with the umbilical vein N2O concentration (r = -0.457 and -0.423, respectively). CONCLUSIONS: The data suggest that placental N2O transfer during Caesarean section is time-dependent and slower compared with maternal N2O uptake. They also suggest that the Apgar score is less affected by N2O administration when the I-D interval is shorter. PMID- 12884990 TI - Comparison of the effects of two intrathecal anaesthetic techniques for transurethral prostatectomy on haemodynamic and pulmonary function. AB - BACKGROUND AND OBJECTIVE: Transurethral prostatectomy is routinely performed under spinal anaesthesia. This technique can cause hypotension, which is particularly undesirable in the elderly. The objective was to compare spinal anaesthesia for transurethral prostatectomy using hyperbaric bupivacaine 15 mg (control group) and hyperbaric bupivacaine 10 mg (limiting spread by maintaining the upright position for 15 min) and fentanyl 25 microg (fentanyl group) in terms of haemodynamic and pulmonary function. METHODS: Thirty ASA I-III patients were randomly selected and underwent spinal anaesthesia with either hyperbaric bupivacaine 15 mg (immediately positioned supine) or hyperbaric bupivacaine 10 mg (upright for 15 min) and fentanyl 25 microg. RESULTS: The greatest changes in mean arterial pressure (P = 0.9), ephedrine requirements (P = 0.8) and mean maximum change in forced vital capacity (P = 0.5) were similar in both groups. CONCLUSIONS: The addition of fentanyl 25 microg to bupivacaine 10 mg and limiting the spread of the block does not improve either haemodynamic or pulmonary function compared with bupivacaine 15 mg in patients undergoing transurethral prostatectomy. PMID- 12884991 TI - Target-controlled infusion of propofol for fibreoptic intubation. AB - BACKGROUND AND OBJECTIVE: In a retrospective study, we examined the suitability of a departmental clinical protocol for anaesthesia induction with target controlled infusion of propofol developed for fibreoptic intubation in spontaneously breathing patients scheduled for outpatient oral surgery at the dental clinic of the Vienna University Hospital. METHODS: Propofol was administered using target-controlled infusion (Diprifusor) at increasing target plasma concentrations starting at 2.5 microg mL(-1). After 10 min, an intravenous dose of alfentanil (5-10 microg kg(-1)) was given for pain reduction. After a further 2 min, the patient was evaluated for response to auditory stimulation. If unresponsive, fibreoptic intubation was performed, otherwise the target concentration was increased by 0.2 microg mL(-1) every 2 min until non responsiveness was attained. RESULTS: Tracheal intubation was successful in all patients without any haemodynamic instability. However, one patient required facemask ventilation for 2 min. No patient was aware of intubation. The plasma concentration required for non-responsiveness was 2.8 +/- 0.4 microg mL(-1) (mean +/- SD). CONCLUSIONS: When using a target-controlled infusion of propofol, fibreoptic intubation can be performed with complete amnesia of the procedure for the patient. However, assisted ventilation of the lungs may be necessary as spontaneous ventilation may cease. PMID- 12884992 TI - Simple method to determine the size of the laryngeal mask airway in children. AB - BACKGROUND AND OBJECTIVE: The size of the laryngeal mask airway in children is determined by the patient's weight. However, in some instances an alternative method may be wanted. The aim was to search for a new method that would be easy to perform at the bedside. METHODS: The size of the laryngeal mask airway was determined in 183 children by choosing the laryngeal mask that best matched the combined widths of the patient's index, middle and ring fingers. The results were compared with the standard method recommended by the manufacturer's weight related guidelines. The patients were classified in different groups depending on the laryngeal mask airway sizes determined by both methods. A kappa coefficient evaluated the agreement between both techniques. RESULTS: The kappa coefficient was 0.81, showing an 'excellent agreement' between both methods. The size was the same for both methods in 142 children (78%). The disagreement between both techniques was only of one size in the remaining 41 patients (22%). In such patients, the weight was a borderline value that would indicate a change in the size of the laryngeal mask airway using the classic method. CONCLUSIONS: This new approach is of valid and practical use in children, particularly as an alternative in those situations where the patient's weight is unknown, such as in emergency situations or in those borderline instances where an alternative measurement would be useful. PMID- 12884993 TI - Effects of diltiazem compared with nicardipine on diaphragmatic fatigability in vivo. PMID- 12884995 TI - Two episodes of bronchospasm during anaesthesia in asthmatic patients. PMID- 12884994 TI - Possible underestimation of the relative incidence of anaphylactic reactions to benzylisoquinoline neuromuscular blocking agents. PMID- 12884996 TI - Temporary pacemakers for non-cardiac surgery. PMID- 12884997 TI - Effects of sevoflurane and isoflurane on the efficacy of rewarming from hypothermic cardiopulmonary bypass. PMID- 12884998 TI - Impossibility of injection through an epidural catheter caused by an incorrect connection of catheter and connector. PMID- 12884999 TI - Cyproheptadine and the treatment of an unconscious patient with the serotonin syndrome. PMID- 12885000 TI - Sometimes my heart goes numb. PMID- 12885001 TI - HIV/AIDS a global epidemic. PMID- 12885002 TI - Prevalence of dental caries and treatment needs among school going children of Pondicherry, India. AB - Dental caries is an important Dental public Health problem. Its high morbidity potential has brought this disease into the focus of dental health professionals. The purpose of the study was to assess the prevalence of dental caries and treatment needs among 5 and 12 years old school children of urban Pondicherry. The study population consisted of 2022 school children of both the sexes, (1009, 5-year-old children) and (1013, 12-year-old children). A simple random sampling method was used to select the schools. Dental caries was assessed by Dentition Status and Treatment Needs described by WHO (1997). The prevalence of dental caries was 44.4% in 5 years age group with 47.4% for males (mean dmft = 1.91 +/- 2.64 S.D.) and 41.1% for females (mean dmft = 1.45 +/- 2.18 S.D.). In 12 years age group the prevalence of dental caries was 22.3% with 20.6% for males (mean DMFT = 0.40 +/- 0.94 S.D.) and 24.1% for females (mean DMFT = 0.55 +/- 1.15 S.D). Evaluation of treatment needs revealed that the greatest need was for one surface restorations followed by two or more surface restorations. It may be concluded that FDI/WHO Oral Health Goals for the year 2000 have been achieved for the ages 5-6 and 12 years in Pondicherry. PMID- 12885003 TI - Mandibular lateral incisor-canine transposition (Mn.12.C): a case report. AB - Tooth transposition is a rare developmental anomaly of the teeth characterized by positional interchange of permanent tooth leading to disintegration in the alignment in the affected segment, shifting of midline and malocclusion of teeth. This article reports a rare case of Mn.12.C anomaly affecting right mandibular arch of a (11 year old) female child causing shifting of midline and crowding. PMID- 12885004 TI - Oral tuberculosis : a case report. AB - Tuberculosis can affect any part of the body and oral cavity remains no exception. The oral lesions of tuberculosis, though not common, are seen in various forms. The purpose of this article is to emphasize the importance of early diagnosis of primary tuberculosis of the mouth, which may be misdiagnosed when the oral lesions are the sole manifestation of the disease. PMID- 12885005 TI - The prevalence of nursing caries in 18 to 60 months old children in Qazvin. AB - The study was carried out to establish the prevalence of Nursing Caries and its relation and caustive factors in children aged 18 to 60 months coming to Health Care Center weighing more than 2.5 kg. A total of 544 children (314 boys and 230 girls) were included in this study. The prevalence of Nursing Caries in girls was 23.5%, in boys 16.5% and in both sexes 19.5%. In affected children dmft was 3.8 times more than the unaffected children and DMFT of their mothers was higher too. Bottle feeding specially during sleep showed statistically significant increase in nursing caries and so did the duration of feeding with mother's milk. The results agree with those obtained in similar populations and showed that the prevalence of nursing caries is anxiously high. PMID- 12885006 TI - Conservative management of a complication of neonatal teeth: a case report. AB - A case report of neonatal teeth in a 4 month old male infant with ulceration on ventral surface of the tongue and its conservative management, is presented. PMID- 12885007 TI - Acidogenic potential of Soya infant formula in comparison with regular infant formula and bovine milk: a plaque pH study. AB - Soya infant formulas are essential for infants intolerant to cow's milk. Soya formulas contains glucose syrup or maltodextrins instead of Lactose, which has led to concern that they are potentially more cariogenic than standard infant formulas containing lactose. The study was conducted to compare the acidogenic effect of Soya Infant formula, infant formula and bovine milk. The study group consisted of 75 children in the age group of 7-10 years. The plaque pH was measured using plaque pH meter, before and after rinsing with different samples for sixty minutes Soya infant formula is found to be more acidogenic than infant formula (with lactose) and bovine milk. This study demonstrates high caries inducing potential of Soya infant formula. Further awareness of the possible role of Soya infant formula in early childhood caries is required. PMID- 12885008 TI - Fused supernumeraries in deciduous dentition: a case report. AB - The occurrence of supernumerary teeth and fusion of two teeth is not a very rare findings but the fusion of two supernumeraries especially in primary is very rare. Reported here, is a case of three supernumerary teeth present in the same region, out of which two of them were fused with each other. Since the interference with the normal eruption of the permanent successor was anticipated, the fused supernumeraries were extracted. PMID- 12885009 TI - Prevalence of supernumerary teeth in deciduous and mixed dentition. AB - Supernumerary teeth are the extra teeth which may have either erupted or unerupted in addition to normal dentition and are seen both in deciduous as well as permanent dentition. The incidence of supernumerary teeth shows more frequent occurrence in permanent dentition than deciduous dentition, affecting both the sexes. The etiology of supernumerary teeth is still unknown and not well understood, but thought to be the result from disturbance during the initiation and proliferation stages of tooth development. In the present study an attempt has been made to asses, its prevalence in deciduous and mixed dentition phase. PMID- 12885010 TI - Acute idiopathic thrombocytopenic purpura in childhood: a case report. AB - Acute thrombocytopenic purpura is the most common of thrombocytopenias of the childhood. Clinical features include petechial lesions on oral mucosa, gingival bleeding and occasionally hemorrhage. A case of acute thrombocytopenic purpura is presented. PMID- 12885011 TI - Ortho-Babinet polarization-interrogating filter: an interferometric approach to polarization measurement. AB - A novel, interferometric, polarization-interrogating filter assembly and method for the simultaneous measurement of all four Stokes parameters across a partially polarized irradiance image in a no-moving-parts, instantaneous, highly sensitive manner is described. In the reported embodiment of the filter, two spatially varying linear retarders and a linear polarizer comprise an ortho-Babinet, polarization-interrogating (OBPI) filter. The OBPI filter uniquely encodes the incident ensemble of electromagnetic wave fronts comprising a partially polarized irradiance image in a controlled, deterministic, spatially varying manner to map the complete state of polarization across the image to local variations in a superposed interference pattern. Experimental interferograms are reported along with a numerical simulation of the method. PMID- 12885012 TI - Symmetry breaking and high-frequency periodic oscillations in mutually coupled laser diodes. AB - We investigate the dynamical behavior of two laser diodes coupled through mutual injection of their optical fields when placed face to face with a small separation between them. We report symmetry breaking in periodic solutions at low coupling rates. In addition, we demonstrate that at higher coupling rates both lasers exhibit very fast periodic oscillations. The system is of practical interest, since it constitutes a tunable all-optical source of microwave oscillations. PMID- 12885013 TI - Laser-induced reorientation effect and ripple structure in dye-doped liquid crystal films. AB - The effects of light-induced reorientation on a homeotropical dye-doped liquid crystal (DDLC) cell are discussed. The photoexcited azo dye Methyl Red (MR) is diffused and adsorbed onto the substrate, thus forming a ripple structure. The adsorbed dye and the laser-induced ripple structure then reorient the liquid crystal molecules and induce a holographic grating. Initially, the liquid-crystal directors are reoriented primarily by the adsorbed dye. However, given a sufficiently large ripple groove amplitude, the torque imposed by the ripple grooves overcomes that which is due to the adsorbed dyes, and the liquid crystals are realigned along the groove direction. PMID- 12885014 TI - New effects in Young's interference experiment with partially coherent light. AB - We analyze the coherence properties of a partially coherent optical field emerging from two pinholes in a plane opaque screen. We show that at certain pairs of points in the region of superposition the light is fully coherent, regardless of the state of coherence of the light at the pinholes. In particular, this result also holds if each pinhole is illuminated by a different laser. PMID- 12885015 TI - Manifestation of the rotational Doppler effect by use of an off-axis optical vortex beam. AB - We report what is to our knowledge the first all-optical detection of the frequency beats between Gaussian and Laguerre-Gaussian LG0(1) modes in their axial superposition, caused by the rotational Doppler effect. The relation between the observable off-axis optical vortex rotation and the rotational frequency shift of the Laguerre-Gaussian component is ascertained. The results can be used as a physical basis for recognition of Laguerre-Gaussian mode spectra along their orbital angular momenta. PMID- 12885016 TI - Opposite-parity orthonormal function expansion for efficient full-vectorial modeling of holey optical fibers. AB - An improved full-vectorial method exploiting the opposite-parity property of eigenmodes based on orthonormal-functions expansion is proposed to solve the wave equation for holey optical fibers. By use of the parity property of eigenmodes in symmetric structures, the number of orthonormal function integrals involved in the calculation is reduced, and the computation efficiency is greatly enhanced. The coupling between the two transverse field components is considered, and both dominant and minor field components can be calculated for the accurate modeling of fiber modes. This method is useful for efficiently modeling holey fibers, especially those with large air holes, in which the coupling effect that is due to refractive-index discontinuities is strong. PMID- 12885017 TI - Incomplete collisions in strongly dispersion-managed return-to-zero communication systems. AB - Incomplete collisions in wavelength-division-multiplexed return-to-zero transmission systems are analyzed by asymptotic methods. Formulas for frequency and timing shifts are obtained. The results agree with direct numerical calculations. PMID- 12885018 TI - Fast phase unwrapping algorithm for interferometric applications. AB - A wide range of interferometric techniques recover phase information that is mathematically wrapped on the interval (-pi, pi). Obtaining the true unwrapped phase is a longstanding problem. We present an algorithm that solves the phase unwrapping problem, using a combination of Fourier techniques. The execution time for our algorithm is equivalent to the computation time required for performing eight fast Fourier transforms and is stable against noise and residues present in the wrapped phase. We have extended the algorithm to handle data of arbitrary size. We expect the state of the art of existing interferometric applications, including the possibility for real-time phase recovery, to benefit from our algorithm. PMID- 12885019 TI - High transmission through waveguide bends by use of polycrystalline photonic crystal structures. AB - A hybrid photonic-crystal structure is presented as a candidate for enhancing transmission through sharp photonic-crystal waveguide bends built on a perforated dielectric slab. This structure, which we refer to as a polycrystalline structure, combines two photonic-crystal lattices. Polycrystalline photonic crystal structures offer the ability to minimize reflections as well as mismatches that a propagating wave might encounter while undergoing a sharp corner or a discontinuity between different waveguide sections. The availability of polycrystalline structures in photonic crystals opens a broad range of possibilities for the development of optical devices. Numerical experiments are performed with two- and three-dimensional finite-difference time domain methods. PMID- 12885020 TI - Self-organization in a multicore fiber laser array. AB - We explain an observed spontaneous transition to the high-brightness, in-phase array state of a seven-core ytterbium-doped fiber laser array [IEEE Photonics Technol. Lett. 13, 439 (2001)]. The responsible mechanism is nonlinear refraction, and either in-phase or antiphase array modes can be selected by control of pump intensity. The phenomenon appears to be robust and scalable. PMID- 12885021 TI - Mutual injection locking of an erbium-doped fiber laser and a fiber-pigtailed Fabry-Perot laser diode. AB - Mutual injection locking of a fiber-pigtailed Fabry-Perot laser diode (FPLD) and a closed-loop erbium-doped fiber-amplifier-based laser (EDFL) system are studied. A single FPLD longitudinal mode can be lasing in the EDFL-FPLD link, with a reduced linewidth of <0.017 nm and an improved sidemode-suppressing ratio of 50 dB. The FPLD's optimized current range is below threshold within 10%, and its required feedback-injecting power is 12.4 microW (corresponding to 0.03% of the EDFL output power). The maximum detuning ranges of the current and temperature of the FPLD that preserve the narrow linewidth and the highest sidemode-suppressing ratio of the EDFL-FPLD link are 2.4 mA and 2.2 degrees C, respectively. The power dissipated by the EDFL-FPLD link is 20% more than that of the free-running EDFL. PMID- 12885022 TI - Optical-fiber-based Mueller optical coherence tomography. AB - An optical-fiber-based multichannel polarization-sensitive Mueller optical coherence tomography (OCT) system was built to acquire the Jones or Mueller matrix of a scattering medium, such as biological tissue. For the first time to our knowledge, fiber-based polarization-sensitive OCT was dynamically calibrated to eliminate the polarization distortion caused by the single-mode optical fiber in the sample arm, thereby overcoming a key technical impediment to the application of optical fibers in this technology. The round-trip Jones matrix of the sampling fiber was acquired from the reflecting surface of the sample for each depth scan (A scan) with our OCT system. A new rigorous algorithm was then used to retrieve the calibrated polarization properties of the sample. This algorithm was validated with experimental data. The skin of a rat was imaged with this fiber-based system. PMID- 12885023 TI - Aberro-polariscope for the human eye. AB - We have developed an aberro-polariscope that simultaneously measures spatially resolved polarization properties and wave-front aberration in a living human eye. The setup consists of an infrared Hartmann-Shack sensor that incorporates a polariscope. A series of four Hartmann-Shack images corresponding to independent polarization states were recorded. The corresponding wave-front aberration was computed from each image. Moreover, from each set of four images spatially resolved (over the pupil plane) parameters of polarization were also determined. This instrument allows useful information on both the optical and the biomechanical properties of the eye to be obtained. PMID- 12885024 TI - In vivo near-infrared spectral detection of pressure-induced changes in breast tissue. AB - A diffuse near-infrared tomography system was used to measure dynamic changes in the absolute optical properties of the human breast that were induced through pressure applied to the tissue surface. Results from five subjects show that absorption and scattering coefficients changed measurably when pressure was increased and that these relative changes correlated with the subjects' body-mass index, indicating that the effect depends on tissue composition. Fitting the absolute absorption and scattering coefficients at six wavelengths to the molar absorption spectra of the three predominant chromophores revealed that both the average total hemoglobin and oxygen saturation increased by 10%, while water concentration decreased by more than 12%. These changes indicate that the pressure-induced variation is likely due to water displacement and vascular volume increase in the region being imaged, for mild application of pressure to the breast. These results suggest that the pressure applied during optical measurements of tissue may alter the tissue physiology, and care should be taken to factor this effect into the design of optical medical instrumentation. In addition, the technique provides a unique approach to measuring tissue elastic changes in vivo in the female breast and may offer a new method for dynamic contrast imaging based on elasto-optical measurements. PMID- 12885025 TI - Measurement of dye diffusion in agar gel by use of low-coherence interferometry. AB - We demonstrate low-coherence interferometry for diffusion measurements. We have measured the diffusion coefficient of a phthalocyanine dye in 1.5% agar gel with a two-wavelength interferometer; one wavelength was matched to the absorption peak of the dye at 675 nm, while the other, 805 nm, was not affected by the dye. The diffusion coefficient of the dye was found by fitting a mathematical model for the interferometer signal to the measured low-coherence interferometry amplitude. A 95% confidence interval for the diffusion coefficient was found to be D = (2.5 +/- 0.2) x 10(-10) m2/s. The influence of speckle averaging and experiment time on the determination of the diffusion coefficient has been studied. The presented technique allows in situ characterization of diffusion in semitransparent media. PMID- 12885026 TI - Functional optical coherence tomography for detecting neural activity through scattering changes. AB - We have demonstrated functional optical coherence tomography (fOCT) for neural imaging by detecting scattering changes during the propagation of action potentials through neural tissue. OCT images of nerve fibers from the abdominal ganglion of the sea slug Aplysia californica were taken before, during, and after electrical stimulation. Images acquired during stimulation showed localized reversible increases in scattering compared with those acquired before stimulation. Motion-mode OCT images of nerve fibers showed transient scattering changes from spontaneous action potentials. These results demonstrate that OCT is sensitive to the optical changes in electrically active nerve fibers. PMID- 12885027 TI - Direct measurement of the voltage sensitivity of second-harmonic generation from a membrane dye in patch-clamped cells. AB - We report what is to our knowledge the first optical imaging of voltage-clamped cells by second-harmonic generation. For the membrane-staining styryl dye di-4 ANEPPS, we determined the sensitivity of second-harmonic generation to be 18%/100 mV at an excitation wavelength of 850 ns. This sensitivity is significantly better than the optimal 10%/100 mV under fluorescence and further establishes the importance of second-harmonic generation for the functional imaging of membrane potential in living cells. PMID- 12885028 TI - Enhanced two-photon biosensing with double-clad photonic crystal fibers. AB - A double-clad photonic crystal fiber was used to improve detection efficiency over a standard single-mode fiber in a two-photon fluorescence detection scheme in which the dye was excited and the fluorescence was detected back through the same fiber. PMID- 12885029 TI - Determination of flow velocity vector based on Doppler shift and spectrum broadening with optical coherence tomography. AB - We describe a technique that uses Doppler optical coherence tomography to estimate accurately the scattering fluid-flow velocity without a priori knowledge of the Doppler angle. Our technique is based on the combined use of the Doppler shift on the interference signal and the Doppler spectrum broadening caused by the particles moving across the probe beam. It is shown that the estimated values of the Doppler angle and average fluid velocity from the experiments agree well with the preset values. PMID- 12885030 TI - Fourier-domain low-coherence interferometry for light-scattering spectroscopy. AB - We present a novel method for obtaining depth-resolved spectra for determining scatterer size through elastic-scattering properties. Depth resolution is achieved with a white-light source in a Michelson interferometer with the mixed signal and reference fields dispersed by a spectrograph. The spectrum is Fourier transformed to yield the axial spatial cross correlation between the signal and reference fields with near 1-microm depth resolution. Spectral information is obtained by windowing to yield the scattering amplitude as a function of wave number. The technique is demonstrated by determination of the size of polystyrene microspheres in a subsurface layer with subwavelength accuracy. Application of the technique to probing the size of cell nuclei in living epithelial tissues is discussed. PMID- 12885031 TI - Optical waveguide sensor for on-line monitoring of bacteria. AB - A grating-coupled planar optical waveguide sensor is presented for sensing of bacteria by evanescent waves. The waveguide design results in increased depth of penetration into the sample volume, which makes it suitable for detecting micrometer-sized biological objects. We tested the sensor's performance by monitoring the adhesion of Escherichia coli K12 cells to the sensor surface. PMID- 12885032 TI - Procedure for retrieving the optical properties of a two-layered medium from time resolved reflectance measurements. AB - A procedure for retrieving the optical properties of a two-layered diffusive medium based on an exact analytical solution of the diffusion equation and on relative multidistance time-resolved reflectance measurements is presented. The method overcomes some limitations of previously developed procedures. Five parameters of the medium have been fitted: the absorption and the reduced scattering coefficients of both layers and the thickness of the first layer. The actual values of the parameters are correctly retrieved by the procedure. The inversion procedure does not require an initial guess for the unknown optical properties, but the starting value for the thickness of the first layer needs to be estimated with an error smaller than 50%. PMID- 12885033 TI - White-light solitons. AB - Optical spatial solitons made from incoherent white light were experimentally observed in 1997 by Mitchell and Segev [Nature (London) 387, 880 (1997)]. We present what is believed to be the first theory describing these solitons and find the characteristic features of their spatiotemporal coherence properties and their temporal power spectrum. PMID- 12885034 TI - Noncritically phase-matched second-harmonic generation in cesium lithium borate. AB - Efficient generation of 236-nm light was demonstrated by use of noncritically phase-matched second-harmonic generation in cesium lithium borate. Noncritical phase matching provided approximately 20x the nonlinear drive for second-harmonic generation than beta-barium borate for 236-nm generation. The 236-nm wavelength is the fourth harmonic of a 946-nm Nd:YAG laser. Phase matching was accomplished at a crystal temperature of -15 degrees C. PMID- 12885035 TI - High-conversion-efficiency optical parametric chirped-pulse amplification system using spatiotemporally shaped pump pulses. AB - High-conversion-efficiency, high-stability optical parametric chirped-pulse amplification is demonstrated with a spatiotemporally shaped pump laser system. Broadband 5-mJ pulses are produced at a 5-Hz repetition rate with a pump-to signal conversion efficiency of 29% and energy stability better than 2% rms. To our knowledge this is the highest conversion efficiency and stability achieved in an optical parametric chirped-pulse amplification system. PMID- 12885036 TI - Pulsed injection-seeded optical parametric oscillator with low frequency chirp for high-resolution spectroscopy. AB - An injection-seeded optical parametric oscillator (OPO), based on periodically poled KTiOPO4 and pumped by a frequency-doubled, nanosecond-pulsed Nd:YAG laser, generates continuously tunable, single-longitudinal-mode, pulsed output at approximately 842 nm for high-resolution spectroscopy. Optical-heterodyne measurements show that the OPO frequency chirp increases linearly with detuning from the free-running (unseeded) OPO frequency and can be maintained as low as 10 MHz. Other factors affecting chirp are identified. PMID- 12885037 TI - X waves generated at the second harmonic. AB - The process of optical frequency doubling in the undepleted regime can lead to the generation of an X-wave envelope with group velocity locked to the pump beam. Its parameters and its angular spectrum are directly related to the zeroth- and first-order dispersive features of the nonlinear process. This constitutes a novel mechanism for spatiotemporal localization of light. PMID- 12885038 TI - Simple method for determining the gain coefficient of a photorefractive polymer film. AB - We propose a simple method for measuring the gain coefficient of two-beam coupling for photorefractive polymer film. To measure the gain coefficient, we attached a phase grating onto the photorefractive polymer film and made an interference pattern between the transmitted and diffracted beams in the photorefractive film by using an illuminating single beam. The gain coefficient values measured at various external fields by our method showed good agreement with those measured by a standard two-beam coupling method. Compared to standard two-beam coupling, this method exhibited fairly improved stability of the output signal to external factors such as human sounds and airstreams. PMID- 12885039 TI - Digital holographic microscope with automatic focus tracking by detecting sample displacement in real time. AB - We propose a new method for focus tracking during the recording of a sequence of digital holograms while the sample experiences axial displacement. Corrected reconstruction distances can be automatically calculated, and well-focused amplitude and phase-contrast images can be obtained for each digitized hologram. The method is demonstrated for inspection of microelectromechanical systems subjected to thermal load. The method can be applied as a quasi-real-time procedure. PMID- 12885040 TI - Silicon-based transmissive diffractive optical element. AB - A silicon nitride (SiNx) membrane diffractive optical element (DOE) designed to exhibit beam-splitting and focusing behavior at visible wavelengths has been fabricated and tested. Since the fabrication process is based on silicon micromachining technology, the DOE is easily integrated with a laser diode chip and a photodiode chip on a silicon substrate to function as the hologram-laser photodiode unit for use in the pickup head of a CD or DVD system. The SiNx film is deposited with low-pressure chemical-vapor deposition and the free-standing membrane is formed by KOH etching. The transmissive DOE showed a high diffraction efficiency (>20% for a binary-phase-level element). The experimental evaluation was in good agreement with the designed and modeled predictions. PMID- 12885041 TI - Grating-enhanced through-wafer optical microprobe for microelectromechanical system high-resolution optical position feedback. AB - We present modeling and experimental results from the use of a 1310-nm-wavelength through-wafer optical microprobe in conjunction with a microstructure grating to monitor the motion of a lateral comb resonator stage. The optical signal that results from shuttle interaction with the microprobe beam exhibits a peak-to valley dynamic range that corresponds to 2-microm microstructure displacement, facilitating submicrometer positional resolution on digitization. This signal was used to achieve microstructure positional feedback and effective microsystem model parameter extraction, which are essential for structure control and model based fault detection. PMID- 12885042 TI - Dense atom clouds in a holographic atom trap. AB - We demonstrate the production of high-density cold 87Rb samples (2 x 10(14) atoms/cm3) in a simple optical lattice formed with YAG light that is diffracted from a holographic phase plate. A loading protocol is described that results in 10,000 atoms per 10 microm x 10 microm x 100 microm unit cell of the lattice site. Rapid free evaporation leads to a temperature of 50 microK and phase space densities of 1/150 within 50 ms. The resulting small, high-density atomic clouds are very attractive for a number of experiments, including ultracold Rydberg atom physics. PMID- 12885043 TI - Self-guided propagation of femtosecond light pulses in water. AB - We report experimental evidence of self-guided propagation of femtosecond laser pulses in water. A light filament induced by 170-fs, 527-nm pulses has a diameter of 60 microm (at the 1/e2 level) and persists over a distance of 20 mm. The filamentary mode is sustained over a wide range of input power, and the energy excess is converted into conical emission. In the time domain, the pulse trapped in a filamentary mode experiences a number of splittings occurring in the early stage of filament formation. PMID- 12885044 TI - Experimental demonstration of phase control of dispersion effects for an ultrashort pulse train propagating in a resonant medium. AB - We present an experiment in which an ultrashort pulse train propagates resonantly through anoptically dense vapor of atomic rubidium. The sequence obtained from a Fabry-Perot interferometer comprises nearly 10 regularly time-delayed and mode locked pulses. We show that a sequence with phase shift phi = 0[2pi] between two successive pulses propagates with important temporal distortion, whereas a sequence with phi = pi[2pi] experiences few propagation effects, thus leading for the first time to our knowledge to the possibility of phase control of dispersion effects for an ultrashort pulse train. PMID- 12885045 TI - Passive mode locking of a Nd:YVO4 laser with an extra-long optical resonator. AB - We describe a mode-locked, diode-pumped Nd:YVO4 laser with a very long optical cavity operating at 1064 nm. High-modulation, InGaAs quantum-well, semiconductor saturable-absorber mirrors were used for passive mode locking, providing a stable train of 13-ps pulses. A novel zero-q-transformation multipass cell provided a variable-length optical cavity as much as 100 m long. The output beam had M2 < 1.1 at average powers of 4.1, 3.9, and 3.5 W at repetition rates of 4.1, 2.6, and 1.5 MHz, respectively. To the best of our knowledge the last of these is the lowest repetition rate ever generated directly from a mode-locked nonfiber solid state laser without cavity dumping. PMID- 12885046 TI - Experimental comparison of three monoclonal antibodies for the class-selective immunoextraction of triazines. Correlation with molecular modeling and principal component analysis studies. AB - The specificity of three immunosorbents (ISs) based on different monoclonal anti triazine antibodies has been characterized by extraction recoveries studies and with step elution experiments. Both indicated that the anti-dichloroatrazine IS is specific of terbutylazine and cyanazine. The anti-atrazine IS is specific of the chlorotriazines, whereas the anti-ametryn IS can trap all the triazines. This confirms the great influence of the hapten design on the specificity of the resulting antibodies, even if the target molecules are small. Moreover, the anti ametryn IS is suitable for class-selective extraction of triazines contained in complex matrices. An approach designed to learn more about the specificity for a group of structurally related compounds of antibodies produced with a given compound is proposed and evaluated. Molecular modeling followed by principal component analysis has been used to obtain distribution maps with the relative position of each immunoconjugate and all the triazines. In all three cases, conclusions on specificity made with the analysis of the maps fit well with the experimental results. Consequently, molecular modeling coupled with principal component analysis seems to be a unique, inexpensive, and rapid tool to select the appropriate hapten providing highly specific or class-specific antibodies according to the given problem. PMID- 12885047 TI - Cyclodextrin-assisted capillary electrophoresis for determination of the cyclic nitramine explosives RDX, HMX and CL-20 comparison with high-performance liquid chromatography. AB - A sulfobutyl ether-beta-cyclodextrin-assisted electrokinetic chromatographic method was developed to rapidly resolve and detect the cyclic nitramine explosives 2,4,6,8,10,12-hexanitro-2,4,6,8,10,12-hexaaza-isowurtzitane (CL-20), octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine (HMX) and hexahydro-1,3,5 trinitro-1,3,5-triazine (RDX) and their related degradation intermediates in environmental samples. Development of the electrophoretic method required the measurement of the aqueous solubility of CL-20 which was determined to be 3.59 +/ 0.74 mg/l at 25 degrees C (95% confidence interval, n=3). The performance of the method was then compared to results obtained from existing high-performance liquid chromatography methods including US Environmental Protection Agency method 8330. PMID- 12885048 TI - Optimization of the class-selective extraction of triazines from aqueous samples using a molecularly imprinted polymer by a comprehensive approach of the retention mechanism. AB - Direct, selective solid-phase extraction of triazines from aqueous samples is presented using a molecularly imprinted polymer (MIP) made with terbutylazine as template molecule. After optimization of the steps of the procedure, 14 triazines including degradation products were studied and satisfactory extraction recoveries were obtained except for thiotriazines. By comparing results obtained with the terbutylazine MIP and a similar non-imprinted polymer, it was determined that retention was achieved via specific interactions except for hydroxyterbutylazine. Selectivity of the extraction procedure was also verified by applying the MIP for the extraction of phenylureas that were not retained on it. The effects of the charge distribution and of molecular volume of the triazines (obtained by molecular modeling) on the selectivity of interactions between the analytes and the MIP were studied. However, when the optimized procedure was applied to real samples, low extraction recoveries were obtained due to strong matrix effects: ion-exchange occurs between the carboxylate groups of the MIP and the ionic species of the sample, that prevents subsequent specific interactions. By introducing an acid wash step, the procedure was successfully applied for the class-selective extraction of triazines from industrial effluent and surface water samples. Finally, increased extraction recoveries were achieved for the polar degradation products of triazines by using a mixed-phase composed of a polymeric sorbent and the MIP. PMID- 12885049 TI - Solid-phase microextraction fibre-water distribution constants of more hydrophobic organic compounds and their correlations with octanol-water partition coefficients. AB - Fibre coating-water distribution constants (Kfw) of more hydrophobic chemicals were determined with six different solid-phase microextraction fibre types (including a polyoctylmethylsiloxane (C8) prototype) by fibre exposure to agitated large-volume water samples lasting several days. The results obtained are comparable with Kfw values obtained under dynamic conditions. Octanol-water partition coefficients (Kow) were measured with the mixture of distributing substances to examine log Kfw-log Kow relationships based on a consistent dataset. Kfw values obtained with liquid polymer fibre coatings correlate reasonably well with the hydrophobicity of the test compounds (with decreasing sensitivity in the following order: 7 microm polydimethylsiloxane (PDMS)>100 microm PDMS>C8>polyacrylate) whereas this is not the case with the PDMS divinylbenzene (DVB) and Carbowax-DVB fibres, partially porous coatings which provide the highest Kfw values for all substances tested. PMID- 12885050 TI - On-site calibration method based on stepwise solid-phase microextraction. AB - A stepwise solid-phase microextraction (SPME) method was developed for on-site calibration of SPME for volatile organic compounds analysis. In this approach, a 75-microm Carboxen-polydimethylsiloxane coated fibre was loaded with a prior to exposure to samples of interest Extraction time for the target analytes can be controlled independently from that of the standard, and the response factors for the target analytes can be adjusted accordingly. A good reproducibility of the response factors for BTEXs (benzene, toluene, ethylbenzene and xylenes) was obtained with stepwise SPME. Satisfactory results were obtained by using this method for quantitative analysis of BTEXs in the air of a gas station when tetrachloroethylene was used as a standard. The introduction of standard via the stepwise SPME procedure makes SPME more useful in field applications. It can be used to detect leaks, contaminations and losses from loading of a standard onto a fibre to introduction of the fibre to an analytical instrument. However, this method cannot be used for compensation of sample matrix effects. PMID- 12885051 TI - Determination of linear alkylbenzensulfonates in aqueous matrices by ion-pair solid-phase microextraction-in-port derivatization-gas chromatography-mass spectrometry. AB - Trace determination (low ng/ml) of linear alkylbenzensulfonates (LASs) in water was achieved by solid-phase microextraction (SPME) of ion-pairs formed with tetrabutylammonium. This ion-pairing reagent served two purposes. First, it allowed the extraction of LAS with the polydimethylsiloxane fiber by counterion association and second, the derivatization of the formed LAS ion pairs in the GC injection port at 300 degrees C to form the corresponding sulfonated butyl esters. The methodology developed allows the isomer specific determination of LAS at low detection limits (0.16-0.8 ng/ml), depending on the alkyl chain lengths of LASs with RSDs of 10-12%. Furthermore, the developed methodology was applied to urban wastewater and sea water and compared with a solid-phase extraction (SPE) method (e.g. C18 and strong anion-exchange sorbent) to obtain concordant values for urban wastewater. Moreover, the developed SPME methodology overcame the procedural blank and matrix-dependent recoveries found in the SPE methodologies at low LAS concentrations. PMID- 12885052 TI - Physico-chemical approach to study organotin sorption-desorption during solid phase microextraction. AB - Solid-phase microextraction (SPME) has become a real alternative to liquid-liquid extraction in the field of speciation of organometallic compounds. Despite the high performance of this preconcentration technique, matrix effects in natural samples can affect the analytical precision. In order to understand the origin of these disturbances and control the extraction step as best as possible, the sorption-desorption behaviour of organotins was studied. In the first part, this paper discusses the analytical problems encountered in the daily use of SPME due to the particular problems observed for phenyltins. The sorption profile of these compounds was modelled using experimental design methodology to confirm the first order kinetics. Desorption of the compounds was also observed after a given time and could not be attributed to competition between organotin compounds. In the same way, butyl- and phenyltins were studied in the presence of humic substances, which acted as representatives of organic matter found in natural samples. These substances drastically decrease the extraction yields, but do not affect the sorption profile of butyl- and phenyltins. PMID- 12885053 TI - Development of a quantification method for the analysis of malodorous sulphur compounds in gaseous industrial effluents by solid-phase microextraction and gas chromatography-pulsed flame photometric detection. AB - A quantification method for malodorous sulphur compounds in gaseous industrial effluents using solid-phase microextraction sampling followed by gas chromatography-pulsed flame photometric detection has been developed. A comparative study showed that polydimethylsiloxane-Carboxen fibre led to sufficient sensitivity to achieve the microg m(-3) human perception levels of the five analytes studied (hydrogen sulphide, methanethiol, ethanethiol, dimethyl sulphide, dimethyl disulphide). However, this coating is known to suffer from competitive adsorption, which may lead to inaccurate quantification. Therefore, external calibration can only be used under a limited range of concentrations, which were determined from Fick's diffusion law. This approach was tested on a real gaseous sample and compared with the standard addition method. Good correlations were found for ethanethiol, dimethyl sulphide and dimethyl disulphide. However, for more volatile sulphur compounds (i.e., hydrogen sulphide and methanethiol), the easy-to-use external calibration could not be applied and standard additions had to be performed for accurate quantification. PMID- 12885054 TI - Determination of methyl tert.-butyl ether and tert.-butyl alcohol in seawater samples using purge-and-trap enrichment coupled to gas chromatography with atomic emission and mass spectrometric detection. AB - A rapid and simple analytical method has been established for the determination of methyl tert.-butyl ether (MTBE) and tert.-butyl alcohol (TBA), in seawater. The method involves purge-and-trap enrichment followed by gas chromatographic (GC) determination. Two different detection systems have been compared: atomic emission detection (AED) and MS (selected ion monitoring mode). Validation parameters and possible matrix effects have been evaluated. The linearity and analytical precision was good with both methods, but limits of detection reached by AED (10 microg l(-1)) were not low enough to evaluate current environmental concentrations. GC-MS detection presented much better sensitivity [limits of detection (LODs) of 0.04 microg l(-1) for MTBE and 0.09 microg l(-1) for TBA] and selectivity, providing a more reliable determination. The analysis of samples collected from various marinas in the south of Spain (Almeria and Malaga) showed, in all cases, detectable concentrations of MTBE that ranged from below LOD to 1842 microg l(-1), depending on the sampling point and time. TBA was also detected in some cases, with concentration levels that ranged from 400 to 600 microg l(-1). These preliminary results should be followed by monitoring programs in coastal waters, in order to establish real levels of presence of MTBE in our coasts and its possible effect on the marine environment. PMID- 12885055 TI - Analysis of 35 priority semivolatile compounds in water by stir bar sorptive extraction-thermal desorption-gas chromatography-mass spectrometry. I. Method optimisation. AB - A multiresidue method for the determination of 35 organic micropollutants (pesticides and polycyclic aromatic hydrocarbons) in water has been optimised using stir bar sorptive extraction (SBSE) and thermal desorption coupled to capillary gas chromatography-mass spectrometry (GC-MS). In the present work, the different parameters affecting the extraction of the analytes from the water samples to the PDMS-coated stir bars and optimisation of conditions affecting thermal desorption are investigated. The optimised conditions consist of a 100-ml water sample with 20% NaCl addition extracted with 20 mm length x 0.5 mm film thickness stir bars at 900 rpm during 14 h at ambient temperature. Desorption is carried out at 280 degrees C during 6 min under a helium flow of 75 ml/min in the splitless mode while maintaining a cryofocusing temperature of 20 degrees C in the programmed-temperature vaporisation (PTV) injector of the GC-MS system. Finally, the PTV injector is ramped to a temperature of 280 degrees C and the analytes are separated in the GC and detected by MS using full scan mode (m/z 60 400). Under the described conditions, the good repeatability, high analyte recoveries and robustness, make SBSE a powerful tool for routine quality control analysis of the selected semivolatile compounds in water samples. PMID- 12885056 TI - Solid-phase microextraction for herbicide determination in environmental samples. AB - Liquid-liquid extraction or solid-phase extraction followed by gas chromatography (GC) or high-performance liquid chromatography are traditional herbicide residue determination methods for environmental samples. Solid-phase microextraction (SPME) is a solventless, fast, and sensitive alternative herbicide residue extraction method that can be applied to numerous environmental matrices. The objective of this paper was to review SPME literature regarding extraction theory, extraction modes, fiber types, and method optimization in conjunction with present and future SPME applications for herbicide determination in environmental samples. PMID- 12885057 TI - Rapid determination of organotin compounds by headspace solid-phase microextraction. AB - Headspace solid-phase microextraction (SPME) followed by gas chromatography (GC) coupled to pulsed flame photometric detection have been investigated for the simultaneous speciation analysis of 14 organotin compounds, including methyl-, butyl-, phenyl-, and octyltins compounds. The analytical process (sorption on SPME fibre and thermal desorption in GC injection port) has been optimised using experimental designs. Six operating factors were considered in order to evaluate their influence on the performances of a SPME-based procedure. The evaluation of accuracy, precision and limits of detection (LODs) according to ISO standards and IUPAC recommendations has allowed the method to be validated. The LODs obtained for the 14 studied organotins compounds are widely sub-ng(Sn) l(-1). The precision evaluated using relative standard deviation ranges between 9 and 25% from five determinations of the analytes at 0.25-125 ng(Sn) l(-1) concentrations. The accuracy was studied throughout the analysis of spiked environmental samples. These first results show that headspace SPME appears really as attractive for organotins determination in the environment and the monitoring of their biogeochemical cycle. PMID- 12885058 TI - Simultaneous determination of "earthy-musty" odorous haloanisoles and their corresponding halophenols in water samples using solid-phase microextraction coupled to gas chromatography with electron-capture detection. AB - Certain haloanisoles present at trace levels cause a large part of earthy-musty off-flavor problems in drinking water. These potent odorous chemicals come mainly through biomethylation of their corresponding halopenols. To enable the investigation of both families of compounds, a method involving solid-phase microextraction (SPME) was developed and the main parameters governing SPME were optimized. This method allows the simultaneous quantification of haloanisoles and halophenols at levels ranging from 1 to 100 or 250 ng/l, with detection limits of about 0.5 ng/l and could be applied to potable as well as raw surface waters. PMID- 12885059 TI - Hollow-fibre liquid-phase microextraction of phthalate esters from water. AB - A simple and efficient liquid-phase microextraction (LPME) technique using a hollow-fibre membrane, in conjunction with gas chromatography-mass spectrometry has been developed for the extraction and analysis of six phthalate esters in water samples. Parameters such as extraction solvent, agitation of the sample, salt addition and extraction time were controlled and optimised. The developed protocol was found to yield a linear calibration curve in the concentration range from 0.02 to 10 microg l(-1) for most target analytes and the limits of detection were in the low microg l(-1) level, ranging between 0.005 and 0.1 microg l(-1). The repeatability of the method varied between 4% and 11%. Under the present experimental conditions, the performance of the method was found comparable to that of solid-phase microextraction (SPME). The advantage of the proposed method over SPME was that it eliminated carry-over of analytes between runs. The applicability of the developed hollow-fibre LPME method and SPME was demonstrated for real water samples. The ability of both microextraction methods to concentrate many organic analytes was demonstrated as both methods allowed the confirmation of the presence of an extra contaminant (ethyl p-ethoxybenzoate) in bottled mineral water samples. PMID- 12885060 TI - Multiple headspace solid-phase microextraction for the quantitative determination of volatile organic compounds in multilayer packagings. AB - The theory of multiple headspace solid-phase microextraction (HS-SPME) and a method based on multiple HS-SPME for the quantitative determination of volatile organic compounds (VOCs) in packaging materials is presented. The method allows the direct analysis of solid samples without using organic solvents to extract analytes. Multiple headspace solid-phase microextraction is a stepwise method proposed to eliminate the influence of the sample matrix on the quantitative analysis of solid samples by HS-SPME. Different amounts of packaging and different volumes of standard solution were studied in order to remove a substantial quantity of analytes from the headspace at each extraction and obtain the theoretical exponential decay of the peak area of the four successive extractions and, thus, the total area was calculated from these four extractions. In addition, two fibres were compared: carboxen-polydimethylsiloxane (CAR-PDMS) and divinylbenzene-carboxen-polydimethylsiloxane (DVB-CAR-PDMS), as they showed differences in the linearity of the exponential decay with the number of extractions depending on the compound. The CAR-PDMS fibre was better for the VOCs with a low molecular mass, whereas the DVB-CAR-PDMS fibre was better for the VOCs with a high molecular mass. Finally, the method was characterised in terms of linearity, detection limit and reproducibility and applied to analyse four multilayer packaging samples with different VOCs contents. PMID- 12885061 TI - Solvent extraction of organic molecules of exobiological interest for in situ analysis of the Martian soil. AB - A solid-liquid extraction method able to perform in situ extraction of organic compounds on Mars is proposed. The extraction efficiency of various organic solvents was tested and compared to that of water. The selected key compounds are molecules of exobiological interest: glycine, alanine, serine, glutamic acid, oxalic acid, benzoic acid, phthalic acid, isophthalic acid, terephthalic acid and 1,3,5-benzenetricarboxylic acid. Among the organic solvents, propanol gives the highest yield of extraction for all the targeted compounds except for benzoic acid. A mixture of propanol and ethyl acetate increases significantly the extraction yield of benzoic acid. The extraction time was considerably reduced (140 h to 15 min) by using sonication. The method is discussed for an easy automation with coupling to an in situ GC-MS space instrument. PMID- 12885062 TI - Rapid determination of polycyclic aromatic hydrocarbons in sewage sludges using microwave-assisted solvent extraction. Comparison with other extraction methods. AB - The presence of toxic organic micropollutants in municipal sewage sludges is a major problem on account of risks associated with the agricultural use of the sludges and therefore maximum tolerance limits are imposed. The aim of our study was to develop a reliable and fast analytical procedure for the determination of polycyclic aromatic hydrocarbons in sewage sludges, using focused microwave assisted extraction. Optimization of the extraction conditions was performed on real matrices. The results of a 2(3) factorial design showed that extraction time was the only influential factor. The selected conditions (30 W, 10 min, 30 ml solvent) were used for real sludges and a certified marine sediment, leading to recoveries of between 56 and 75%. Results of an interlaboratory test confirmed these values. Finally, the technique was compared to traditional techniques (Soxhlet, sonication), and the more recent pressurized liquid extraction technique. Focused microwave-assisted extraction remains an attractive alternative technique due to its rapidity, the low solvent volume required and its moderate investment cost. PMID- 12885063 TI - Determination of musk compounds in sewage treatment plant sludge samples by solid phase microextraction. AB - Headspace solid-phase microextraction, followed by GC-MS analysis is presented as a suitable technique for the determination of musk compounds in sewage treatment plant sludge. Five polycyclic musks (celestolide, phantolide, traseolide, galaxolide and tonalide) and four nitro musks (musk xylene, musk moskene, musk tibetene and musk ketone) were considered in the optimisation of the analytical method. The influence of extraction temperature, fibre coating, agitation, pH and salting out on the efficiency of the extraction along with the extraction kinetics were studied. An extraction temperature of 100 degrees C and sampling the headspace over the stirred sludge sample using polydimethylsiloxane divinylbenzene as fibre coating lead to effective extraction. The method proposed is very simple and yields high sensitivity, good linearity and repeatability for all the analytes with limits of detection at the sub-ng/g level. The total analysis time, including extraction and GC analysis, was only 40 min, and no manipulation of the sample was required. PMID- 12885064 TI - Use of solid-phase microextraction coupled with gas chromatography for the determination of residual solvents in pharmaceutical products. AB - The aim of this work was to prove that solid-phase microextraction coupled with gas chromatography could be used for the determination and quantification of residual solvents in drugs. Four solvents were selected for the experiments: ethanol, cyclohexane, triethylamine and pyridine, together with a model powdered drug substance. Several kinds of fibers, together with the extraction mode, were evaluated to determine the most appropriate one for the simultaneous extraction of the four solvents. The most promising conditions were obtained with the Carboxen-polydimethylsiloxane fiber in the headspace of the aqueous solution that contained the dissolved powder. A concentrated phosphate buffer was added to the aqueous solution to set the pH at 9.6 in order to enable good extraction of triethylamine, and the optimum extraction time was experimentally determined. A multi-criteria optimization was also carried out by means of design of experiments to optimize remaining parameters: the extraction temperature was set at 40 degrees C, the ionic strength at 1.77 mol (l-1) and the volume of the aqueous solution at 7.2 ml. The method of standard additions was used for quantitative analysis. Its performance was evaluated and validated: the pooled RSD was around 15%, the limits of detection were all of the ppb level and the method was both accurate and linear. PMID- 12885065 TI - What makes a good leader? PMID- 12885066 TI - Using failure mode and effects analysis to improve patient safety. AB - Failure mode and effects analysis (FMEA) (ie, prospective risk analysis) involves close examination of high-risk processes to identify needed improvements that will reduce the chance of unintended adverse events. This risk assessment process is used in other industries (ie, manufacturing, aviation) to evaluate system safety. Health care organizations now are using it to evaluate and improve the safety of patient care activities. The FMEA process promotes systematic thinking about the safety of patient care processes (ie, what could go wrong, what needs to be done to prevent failures.) The steps of the FMEA process are described and applied to a high-risk perioperative process. PMID- 12885067 TI - Incident reports--their purpose and scope. AB - Accidents in the health care setting may be inevitable, but their frequency can be decreased with a dedicated focus on patient safety. Risk reduction naturally flows from a positive approach to risk containment and control that includes learning from past errors. Part One of this two-part series on incident reports talks about identifying and correcting errors, which results in decreased harm to patients and personnel, decreased facility risk liability and regulatory sanctions, and less negative publicity. Although it may be human nature to make mistakes, it also is human nature to create solutions, identify alternatives, and meet future challenges. PMID- 12885068 TI - Identifying lost surgical needles using radiographic techniques. AB - Many surgical procedures involve the use of very small surgical needles. In the event that a postoperative needle count is incorrect, traditionally an x-ray has been taken to find the needle. Little data exist to recommend the smallest surgical needle that can be identified with conventional radiographic techniques and the optimum technique for finding lost surgical needles. In this quality assurance project x-rays of various size surgical needles were taken. The smallest seen by the majority of observers was 17 mm. The radiographic technique of choice for optimum detection of lost surgical needles was imaging with a mobile image intensifier. PMID- 12885069 TI - Heparin-induced thrombocytopenia--recognition and treatment. AB - Heparin, an important agent for the prophylaxis and treatment of venous and arterial thromboembolism, also can cause heparin-induced thrombocytopenia (HIT). This medication-induced immune-mediated syndrome is characterized by thrombocytopenia and life- or limb-threatening thrombotic events. The initial presentation of HIT is a decrease in platelet count with or without thrombosis. Heparin-induced thrombocytopenia frequently is overlooked in hospital settings, and it must be distinguished from other causes of thrombocytopenia. New therapies include the use of direct thrombin inhibitors argatroban and hirudin. PMID- 12885070 TI - A guide to developing a systematic review. AB - Systematic reviews are used to gather information from a variety of sources and distill information based on best evidence synthesis, while also excluding poor quality studies via explicit validity checks. This article provides a roadmap to conducting a systematic review with the Cochrane Collaboration and uses an example of a completed systematic review. The literature is drawn upon to provide rationale for the processes undertaken. The aim of the sample systematic review was to identify and review all randomized controlled trials evaluating disposable surgical face masks worn by surgical team members during clean surgery to prevent postoperative surgical wound infection. PMID- 12885071 TI - Nursing abroad stirs the soul. AB - After establishing herself in her career as a perioperative nurse and ensuring her grown children were settled, this author pursued her dream of working abroad as a nurse. She chose to work in Pakistan, which is where her father was born. Her experiences there gave her a new appreciation for the help that perioperative nurses can offer to the citizens of developing countries. PMID- 12885072 TI - Withdrawing consent after a procedure has begun. PMID- 12885073 TI - Keeping patients safe from infection. PMID- 12885074 TI - Determination of herbicides, including thermally labile phenylureas, by solid phase microextraction and gas chromatography-mass spectrometry. AB - A method for the determination of 10 herbicides, including thermally unstable compounds, has been developed. The method uses solid-phase microextraction (SPME) with a polyacrylate fibre. Separation, identification and quantification were accomplished with gas chromatography-mass spectrometry. The herbicides chosen belong to different chemical groups and were alachlor, atrazine, chlorotoluron, diclofop, diflufenicam, ethofumesate, isoproturon, linuron, terbutryn and trifluralin. In the present work we studied the chromatographic behaviour of three phenylureas as a function of the medium and injection mode employed. The compounds generated as a function of the solvent used in direct injection of the phenylureas (ethyl acetate, methanol and methanol-water) and those obtained when injection was accomplished using the polyacrylate fibre were determined. The results allow us to propose a method for the determination of stable and thermally unstable herbicides as long as a preconcentration step involving SPME is carried out. In the proposed method, the limits of detection varied between 0.02 microg/l for ethofumesate and 0.11 microg/l for chlorotoluron. The method was applied to the determination of these herbicides in surface and ground water samples, performing quantification by standard addition calibration. The contents of chlorotoluron and atrazine found were significantly equal to those obtained using HPLC after a preconcentration stepwith styrene-divinylbenzene sorbents. PMID- 12885075 TI - Direct continuous supercritical fluid extraction as a novel method of wine analysis. Comparison with conventional indirect extraction and implications for wine variety identification. AB - Direct supercritical fluid extraction (SFE) of wines with carbon dioxide was compared to SFE of the sorbent used for solid-phase extraction of the same wine samples (SPE-SFE). Compared to SPE-SFE, the direct SFE results in amore specific and representative gas chromatographic fingerprint of the wine sample. The multivariate statistical processing of the direct SFE-GC data provides a clear cut and sharp discrimination among the individual wine varieties while the discrimination based on the SPE-SFE-GC data is relatively poor. This finding reflects the adverse effects of additional analyte-sorbent interactions and sorption/desorption steps involved in SPE-SFE. PMID- 12885076 TI - Information theory applied to chromatographic fingerprint of herbal medicine for quality control. AB - At present, the construction of chromatographic fingerprints plays an important role in the quality control of complex herbal medicines. In this work, information theory was applied to obtain chromatographic fingerprints with good performance. Moreover, according to the characteristics of the chromatographic fingerprints obtained, some modifications of the calculation of the information content were conducted. In comparison with the information content from several chromatographic fingerprints obtained, reliable chromatographic fingerprints with a high separation degree and uniform concentration distribution of chemical components could be determined. The successful application of information theory with modification to simulated chromatographic fingerprints together with real herbal medicines such as Rhizoma chuanxiong and Ginkgo biloba from different sources demonstrated clearly that the proposed method to determine chromatographic fingerprints was reasonable and reliable and it was user friendly. Chromatographic fingerprints determined with high separation degrees and uniform concentration distribution of chemical ingredients might also chemically represent characteristic components of herbal medicines for quality control. PMID- 12885077 TI - Effect of temperature on pH measurements and acid-base equilibria in methanol water mixtures. AB - The knowledge of the acid-base equilibria in water-solvent mixtures of both common buffers and analytes is necessary in order to predict their retention as function of pH, solvent composition and temperature. This paper describes the effect of temperature on acid-base equilibria in methanol-water solvent mixtures commonly used as HPLC mobile phases. We measured the delta-correction parameter (delta = sw pH - ss pH = Ej - log sw(gamma)oh) between two pH scales: pH measured in the solvent concerned and referred to the same standard state, ss pH, and the pH measured in that solvent mixture but referred to water as standard state, sw pH, for several methanol compositions in the temperature range of 20-50 degrees C. These determinations suggest that the delta-term depends only on composition of the mixture and on temperature. In water-rich mixtures, for which methanol is below 40% (w/w), delta-term seems to be independent of temperature, within the experimental uncertainties, whereas for methanol content larger than 50% (w/w) the delta-correction decreases as temperature increases. We have attributed this decrease to a large increase in the medium effect when mixtures have more than 50% methanol. The pKa of five weak electrolytes of different chemical nature in 50% methanol-water at 20-50 degrees C are presented: the effect of temperature on pKa was large for amines, pyridine and phenol, but almost no dependence was found for benzoic acid. This indicates that buffers can play a critical role in affecting retention and selectivity in HPLC at temperatures far from 25 degrees C, particularlyfor co-eluted solutes. PMID- 12885078 TI - Adsorption of polyethylene standards from decalin on liquid chromatography column packings. AB - Linear polyethylene and isotactic polypropylene standards were injected into columns which contained MFI (SH-300 and silicalite) or faujasite (CBV-780) type zeolites. 1,2,4-Trichlorobenzene, cyclohexanone, 2-ethyl-hexanol, decalin and tetralin were used as mobile phases at 140 degrees C. It was found that polyethylene is fully retained on zeolite SH-300 when decalin is used as a mobile phase. Moreover, polyethylene is partially retained on zeolite SH-300 from tetralin and from 1,2,4-tichlorobenzene, on silicalite from decalin and in a very small extent on zeolite CBV-780 from decalin. Using all other solvents, polyethylene and polypropylene were not retained in any of the columns tested. This is the first experimental observation of polyethylene adsorption from a solvent on a chromatographic stationary phase. PMID- 12885079 TI - Application of diaza-18-crown-6-capped beta-cyclodextrin bonded silica particles as chiral stationary phases for ultrahigh pressure capillary liquid chromatography. AB - Two bonded chiral stationary phases (CSPs), 8-aminoquinoline-2-ylmethyl- and 8 aminoquinoline-7-ylmethyl-diaza-18-crown-6-capped [3-(2-O-beta-cyclodextrin)-2 hydroxypropoxy]propylsilyl silica particles (non-porous, 1.5 microm), have been prepared and evaluated using capillary liquid chromatography at high pressures (> or = 8000 p.s.i.). High column efficiency (up to 400 000 plates m(-1)) was achieved for chiral separations. These CSPs with two recognition sites, i.e. substituted-diaza-18-crown-6 and beta-cyclodextrin combined with high chromatographic efficiency provide good resolution of a variety of enantiomers and positional isomers in relatively short times under reversed-phase conditions. After inclusion of a Ni (II) ion from the mobile phase, the positively charged crown ether-capped beta-cyclodextrin facilitates specific static, dipolar, and host-guest complexation interactions with solutes. PMID- 12885080 TI - Synthesis of micron diameter polybutadiene-encapsulated non-porous zirconia particles for ultrahigh pressure liquid chromatography. AB - In this study, 1-microm diameter polybutadiene-encapsulated non-porous zirconia particles were synthesized, slurry packed into 50-microm I.D. fused-silica capillary columns, and evaluated using ultrahigh pressure liquid chromatography. The dependencies of column efficiency and solute retention factor on pressure were investigated. Efficiencies as high as 280000 plates per meter were obtained for the separation of anti-inflammatory drugs at a pressure of 1351 MPa. Comparing the reversed-phase behavior of the polybutadiene-encapsulated non porous zirconia with octadecylsilane bonded non-porous silica, greater selectivity was found using the zirconia-based material for the applications reported in this study. The encapsulated non-porous zirconia particles demonstrated excellent thermal stability in the separation of polycyclic aromatic hydrocarbons at a temperature of 100 degrees C and a pressure of 1351 MPa. PMID- 12885081 TI - On-line high-performance liquid chromatography-mass spectrometric detection and quantification of N-acylhomoserine lactones, quorum sensing signal molecules, in the presence of biological matrices. AB - A protocol using reversed-phase liquid chromatography coupled with positive-ion electrospray ionization and ion trap mass spectrometry is described for the identification and quantification of N-acylhomoserine lactones (HSLs) in crude cell-free supernatants of bacterial cultures. The HSLs are produced by gram negative bacteria and act as intercellular signals inducing density-dependent gene expression. Compared with the multi-step procedures previously reported, which included chemical extraction, purification and the use of Escherichia coli HSL biosensors, this on-line LC-MS-MS method is fast and detects 11 HSLs. Its speed and robustness allow the analysis of a large number of samples without loss of performance (no signal variation for a control sample after 90 chromatographic injections). The selectivity is based on the MS-MS fragment ions of the molecular [M+H]- ions and on their relative intensities. For quantification, the m/z 102 ion, specific for the lactone ring and detected with a good signal-to-noise ratio, allows low detection limits even in complex matrix samples (0.28 up to 9.3 pmol). Moreover, this method allows the quantification of 11 HSLs whatever their chemical structure, substituted or not. The protocol was applied to Vibrio vulnificus, a marine bacterium. Six HSLs were detected and quantified with relative standard deviations for repeatability of < 10%. PMID- 12885082 TI - Determination of 1-deoxynojirimycin in Morus alba L. leaves by derivatization with 9-fluorenylmethyl chloroformate followed by reversed-phase high-performance liquid chromatography. AB - A rapid and reliable method suitable for assays of a large number of Morus alba leaves for 1-deoxynojirimycin (DNJ) has been developed. DNJ in 0.1 g of freeze dried leaves was double-extracted in 10 mL of aqueous 0.05 M HCl by vortexing for 15 s at room temperature, derivatized with 9-fluorenylmethyl chloroformate (FMOC Cl), and analyzed by reversed-phase high-performance liquid chromatography (RP HPLC) equipped with a fluorescence detector. The double extraction recovered > 99% of extractable DNJ from the leaves. Stabilization of FMOC-derivatized DNJ (DNJ-FMOC) was achieved by diluting the reactant with aqueous acetic acid after derivatization. DNJ-FMOC was stable for at least 16 days under acidic conditions at room temperature (24 degrees C). Linearity ranged between 0.3 and 30 microg mL(-1). The intra- and inter-day precision for DNJ-spiked biological samples was between 0.6 and 1.8% and between 3.7 and 4.5%, respectively. PMID- 12885083 TI - Molar mass characterization of cationic methyl methacrylate-ethyl acrylate copolymers using size-exclusion chromatography with online multi-angle light scattering and refractometric detection. AB - Size-exclusion chromatography (SEC) combined with online multi-angle light scattering (MALS) and refractometric (RI) detection has been employed for the molar mass characterisation of water-insoluble cationic methyl methacrylate-ethyl acrylate copolymers (Eudragit RS and RL). Due to their positive charge, cationic polymers are particularly difficult to separate on a SEC column, in worst cases being completely adsorbed on the oppositely charged packing material. This work has examined how a careful addition of salt (LiCl) to the copolymer solution in ethanol decreases the electrostatic interactions, clearly seen as a decrease in elution volume from the SEC column as well as an improved recovery. At a certain level of ionic strength, typically about 50 mM, the copolymer recovery from the SEC column reached 100% and molar mass distributions corresponding to the complete sample could be obtained. The combined MALS/RI detection gives the opportunity to measure the absolute molar mass independent of recovery and retention. Thus, in this study, it turned out to be a favourable tool for tracing the changes in elution behaviour of the charged copolymer as the ionic strength was increased. PMID- 12885084 TI - Fungal metabolite screening: database of 474 mycotoxins and fungal metabolites for dereplication by standardised liquid chromatography-UV-mass spectrometry methodology. AB - A standardised LC-UV-MS micro-scale method for screening of fungal metabolites and mycotoxins in culture extracts is presented. The paper includes data for detection and dereplication of > 400 fungal metabolites to facilitate detection and identification when standards are not available. The data also shows the types of components that can be analysed by positive electrospray (ESI+) mass spectrometry (MS) along with common fragments and adducts of these, as well as giving suggestions on whether UV or ESI+-MS methods should be used. Examples of dereplication of penitrems and macro-cyclic ichothecenes, and detection of several novel compounds are shown. This was done by UV spectroscopy combined with accurate mass determination of adduct and fragment ions obtained by high resolution orthogonal time-of-flight MS. PMID- 12885085 TI - Procedure for determination of trace ions in boric acid by matrix volatilization ion chromatography. AB - A method for determination of anions and cations in boric acid is proposed by matrix volatilization. The boric acid matrix was eliminated as trimethyl borate ester in a vapour phase matrix elimination (VPME) system using a mixture of glycerol-methanol. In this VPME system, in situ reagent purification, sample decomposition and digest evaporation were achieved in a single step. Trace anions were separated on anion-exchange column (IonPac AS17) by an isocratic elution with 15 mM sodium hydroxide and the cations on a cation-exchange column (IonPac CS12) by 20 mM hydrochloric acid as eluents. Method detection limits (3sigma) for most ions ranged from 0.3 to 8 ng/g (ppb). Recovery experiments combined with comparison of data obtained by other methods were employed to verify the accuracy of the proposed method. Application of the method to determine trace levels of anions like acetate, oxalate, sulfate, phosphate and cations such as lithium, sodium, potassium, magnesium and calcium in two highly pure grades of boric acid using ion chromatography is demonstrated. PMID- 12885086 TI - Sonication effect on cellular material in sedimentation and gravitational field flow fractionation. AB - Sonication procedures are generally used prior to field flow fractionation (FFF) separation in order to produce suspensions without aggregates. Yeast cells manufactured in active dry wine yeast (ADWY) were placed in an ultrasound water bath in order to disrupt possible clumps and to obtain a single-cell suspension to be used in optimal conditions during fermentation processes. In order to determine whether this sample preparation procedure meets absolute needs, different yeast samples before and after sonication were analysed by two field flow fractionation techniques. It is shown that 2 min of sonication in the sample preparation process is sufficient to obtain an optimal dispersion of the yeast cells, that is, without critical percentage of aggregates. To demonstrate this effect, photographs of the yeast cell suspensions were performed with non sonicated and sonicated yeast sample dispersion. The resulting data are compared with the elution profiles obtained from the two different FFF techniques. It is demonstrated that fractogram profiles prove the effectiveness of sonication methodologies. PMID- 12885087 TI - Mosaic increments for predicting the gas chromatographic retention data of the chlorobenzenes. AB - The chlorinated organic compounds are very important from the point of view of the chemical industry and environmental protection, and therefore the gas chromatographic analysis of these compounds is very interesting for analytical chemists. In this paper we studied the relationship between the molecular structure and gas chromatographic retention on several stationary phases having different polarity and at several temperatures of benzene and 12 chlorobenzene compounds as model compounds. A coding system involving primary (mosaic increments) and secondary (bond increments)calculation methods was developed. The retention indices of benzene and the chlorobenzenes calculated on HP-5 at 120 degrees C shows a better performance of the mosaic increments (average absolute deviation delta of 1.7 retention index units) compared with the bond increments (delta = 11.7 retention index units). Retention factors, k, calculated with mosaic increments for chlorobenzenes on SPB-1 and WAX-10, at 140 degrees C, yield average relative errors of epsilon = 0.9 and 3.5%, respectively. Therefore, the presented paper provides a new possibility for precalculation of the retention data. PMID- 12885088 TI - Quantitative gas chromatography-olfactometry. Analytical characteristics of a panel of judges using a simple quantitative scale as gas chromatography detector. AB - Some questions related to the ability of gas chromatography-olfactometry (GC-O) to provide quantitative measurements of the concentration of a given odorant in an extract are explored and discussed. A panel of eight individuals has been used to evaluate the intensity of 15 odorants present at different concentrations. The use of very simple scales, such as a 0-3 scale, makes it possible to build calibration graphs based on the different stimulus-response models (Fechner, Stevens, Hill) and, with a proper calibration, up to nine different concentration levels can be discriminated by an eight-judge panel. The signal shows a good long term stability, and its precision varies between 3.7 and 8% of the whole scale, with 5.7% as average. Sensitivity is extremely dependent on the compound: in the best case a concentration change of 20% can be detected, while in the worst, concentrations must differ more than one order of magnitude. In average, concentrations must differ between 2 and 4.7 times (including calibration error) or between 1.2 and 2.3 (excluding calibration error). The performance of the different judges, the effect of the close elution of two odors and the benefits derived from the use of more complex scales (7-points) are briefly discussed from the perspective of the analytical performance of GC-O methods. PMID- 12885089 TI - Decoding of complex isothermal chromatograms recovered from space missions. Identification of molecular structure. AB - A chemometric approach, based on the study of the autocovariance function, is described to study isothermal GC chromatograms of multicomponent mixtures: isothermal GC analysis is the method of choice in space missions since it is, to date, the only method compatible with flight constraints. Isothermal GC chromatograms look inhomogeneous and disordered with peak density decreasing at higher retention times: a time axis transformation is proposed to make retention an homogeneous process so that CH2 addition in terms of an homologous series yields a constant retention increment. The time axis is transformed into a new scale based on the retention times of n-alkanes, as they are the basis of the universal Kovats indices procedure. The order introduced into the chromatogram by retention time linearization can be simply singled out by the experimental autocorrelation function (EACF) plot: if constant inter-distances are repeated in different regions of the chromatogram, well-shaped peaks are evident in the EACF plot. By comparison, with a standard mixture it is possible to identify peaks diagnostic of specific molecular structures: study of the EACF plot provides information on sample chemical composition. The procedure was applied to standard mixtures containing compounds representative of the planetary atmospheres that will be investigated in the near future: in particular, those related to Titan's atmosphere (Cassini-Huygens mission) and cometary's nucleus (Rosetta mission). The employed experimental conditions simulated those applied to GC instruments installed on space probes and landers in space missions. The method was applied to two specific investigations related to space research, i.e., a comparison of retention selectivity of different GC columns and identification of the chemical composition of an unknown mixture. PMID- 12885090 TI - Calibration system and analytical considerations for quantitative sesquiterpene measurements in air. AB - Sesquiterpenes (C15H24, SQT) are semi-volatile organic compounds emitted from vegetation and are of interest for air quality considerations because of their suspected contribution to the formation of secondary aerosol. This article investigates the application of a capillary diffusion method for the generation of standard atmospheres of 16 SQT and four other related semi-volatile compounds. This instrument subsequently has been used in the testing of analytical materials, protocols and calibration of air sampling methods. SQT DB-1 retention indices, vapor pressures at 25 and 75 degrees C, and diffusion coefficients were determined. A quantitative, on-line GC method yielded improved results (median relative standard deviation of 5.0-6.1%) for the diffusion rate determination in comparison to a gravimetric approach (median relative standard deviation 18%). The GC method also allowed identifying errors in the gravimetric method stemming from residual solvent evaporation, impurities, and chemical analyte losses. Stainless steel, glass, nickel and PTFE tubing that were tested for transfer lines and a sampling loop had to be kept at temperatures in excess of approximately 110 degrees C in order to prevent significant analytical errors from the stickiness of SQT to these materials. In addition to SQT analysis, results from this research provide general guidelines for gas-phase analysis of related compounds in the C14-C16 volatility range. PMID- 12885091 TI - Coupling development and elution, a new thin-layer chromatography technique. AB - Three methods of coupling development and elution were studied in this paper. (1) A new mode of solvent supplementation and eluate collection was developed for descending development. By using a new distributor and collector in descending development, components can be separated and eluted continuously. (2) The same effect can be realized with a slope distributor [Su et al., J. Planar Chromatogr. 14 (2001) 203] and a collector by horizontal development. (3) In-situ elution can be used to treat a developed silica plate, which can elute the separated components to the receptor without scraping them off. These three methods can be used individually, and the in-situ elution can be used with other modes of development. PMID- 12885092 TI - Quantitative structure-property relationship studies of migration index in microemulsion electrokinetic chromatography using artificial neural network. AB - Artificial neural networks (ANNs) were successfully developed for the modeling and prediction of migration indices of the 53 benzene derivatives and heterocyclic compounds in microemulsion electrokinetic chromatography. The selected descriptors that appear in multiple linear regression models are: 3D MoRSE signal 25 unweighted, 3D-MoRSE signal 19 weighted by atomic Sanderson electronegativity, R maximal autocorrelation index lag 1 weighted by atomic mass (R1M+), R maximal autocorrelation index lag 2 weighted by polarizability (R2P+) and average atomic composition index. These descriptors were used as inputs for generated 5-4-1 networks. After training and optimization of the ANN parameters it was used to prediction of migration index of the test set compounds. The results obtained using ANNs were compared with the experimental values as well as with those obtained using regression models and showed the superiority of ANNs over regression models. PMID- 12885093 TI - Enantioseparation of novel chiral heterometal tetrahedral clusters by high performance liquid chromatography. AB - A series of novel chiral heterometal tetrahedral clusters have firstly been separated on cellulose tris-(3,5-dimethylphenylcarbamate) stationary phase by high-performance liquid chromatography using n-hexane as the mobile phase containing different alcohols as modifiers. The effects of concentration and structures of alcohols in the mobile phases and structures of chiral heterometal tetrahedral clusters on enantioseparation were investigated. The results showed that the concentration and structures of alcohols had large effects on enantioseparation. It also was found that both the metal in the tetrahedral core and the ligand coordinated to the atom in tetrahedral core had significant effects on their chromatographic behavior. PMID- 12885094 TI - Ion chromatographic method for the simultaneous determination of nitrite and nitrate by post-column indirect fluorescence detection. AB - This short paper highlights the suitability of ion chromatography with post column indirect fluorescence detection to determine simultaneously nitrite and nitrate based on the quenching of tryptophan native fluorescence. The method uses an enhanced fluorescence mobile phase containing tryptophan and detects the suppression of fluorescence of the mobile phase due to the elution of the target ions. The phenomenon of fluorescence quenching of tryptophan is highly induced by the presence of phosphate ions. The quenched fluorescence intensity exhibits concentration dependence in the range 1-25 mg/l and 3-65 mg/l for nitrite and nitrate, respectively. The relative standard deviation for five replicates of a standard solution containing a mixture of 5 mg/l of nitrite and 10 mg/l of nitrate lies around 2.8%. This simple coupling technique results in a relatively sensitive, fast, and accurate method, allowing for both qualitative and quantitative analysis of nitrite and nitrate. The method can easily be implemented to real samples such as foodstuffs, fertilizers and soils and is proven to be precise and accurate when compared with reference methods. PMID- 12885095 TI - Efficacy and safety of 4-aminopyridine in patients with long-term spinal cord injury: a randomized, double-blind, placebo-controlled trial. AB - OBJECTIVES: To study the efficacy and safety of 4-aminopyridine (4-AP), and to document sensorimotor changes after discontinuation of the drug in patients with long-term spinal cord injury. DESIGN: Randomized, double-blind, placebo controlled trial. SETTING: Clinical research unit. PATIENTS: Twenty-seven patients with long-term spinal cord injury. INTERVENTION: Patients were randomized to receive either oral 4-AP 5 mg/day, which was increased by 5 mg/week to a maximum dosage of 30 mg/day, or placebo for 12 weeks. They switched to the opposite treatment for the next 12 weeks. MEASUREMENTS AND MAIN RESULTS: Twenty five patients finished the study. The results from the first 12 weeks were used to test efficacy. Positive gains in motor function, sensation, and independence occurred more frequently in patients receiving 4-AP (69%) than those receiving placebo (46%). Significant functional improvement was also noted in those treated with 4-AP (chi2, p=0.042). When each evaluation scale was considered separately, significant improvement was seen only in motor function (4-AP 92% vs placebo 46%, Fisher exact test, p=0.03). Persistent effects of the drug were assessed at week 24 in the group that initially received 4-AP. A persistent, significant 4-AP effect was observed in evaluations of sensation and independence (67% and 83% of patients, respectively; Wilcoxon signed rank test, p=0.032 and 0.042, respectively). Fourteen (56%) patients had 26 adverse reactions. One moderate adverse reaction--posterior tibial artery vasospasm--and 25 mild adverse reactions, such as dry mouth, dizziness, nausea, gastritis, oral and peripheral paresthesia, resolved adequately. Six (24%) patients experienced transitory alterations of enzyme levels (alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, and creatine kinase) and thrombocytopenia. CONCLUSION: Patients who received 4-AP showed significant improvement in motor function, and a persistent effect on sensation and independent function occurred. The drug is safe; however, after starting 4-AP therapy, patients must be carefully monitored for the possible occurrence of peripheral vasospasm. PMID- 12885096 TI - Effects of didanosine formulations on the pharmacokinetics of amprenavir. AB - STUDY OBJECTIVES: To determine the effects of concurrent, single doses of didanosine (both buffered and encapsulated enteric-coated bead formulations) on amprenavir steady-state pharmacokinetics, and to determine the effect of staggered dosing of the buffered formulation. DESIGN: Two-period, single sequence, prospective, open-label drug interaction study with a 10-day washout interval. SETTING: Clinical research unit. SUBJECTS: Sixteen healthy volunteers without human immunodeficiency virus infection. INTERVENTION: Amprenavir 600 mg twice/day was given for the first 4 days of each treatment period, with 12-hour pharmacokinetic evaluations conducted on the last 2 days of each period. Amprenavir was administered according to the following sequential treatments (all fasting): amprenavir alone, concurrent with buffered didanosine, 1 hour before buffered didanosine, and concurrent with the encapsulated enteric-coated bead formulation of didanosine. MEASUREMENTS AND MAIN RESULTS: Plasma was collected 0, 1, 2, 3, 4, 6, 8, and 12 hours after dosing and assayed for amprenavir by using high-performance liquid chromatography. Noncompartmental pharmacokinetic parameters were determined. Geometric mean ratios for each treatment relative to amprenavir alone were determined and reported with 90% confidence intervals (CIs). No significant trends were noted in predose concentrations measured during either period. Area under the concentration-time curve during one 12-hour dosing interval (AUC12) was found to be bioequivalent for all treatments. Peak drug concentration (Cmax) was reduced by 15% on average with concurrent administration of buffered didanosine, and bioequivalence was not demonstrated for this parameter. For concurrent enteric-coated didanosine, geometric mean ratios for Cmax and AUC12 were 0.93 and 0.94, respectively. For buffered didanosine given 1 hour after amprenavir, geometric mean ratios were 1.06 and 1.10 for the same parameters, respectively. No differences were observed in 12-hour concentration (C12) with concurrent administration of buffered or enteric-coated didanosine. CONCLUSION: Amprenavir AUC12 and C12 are not significantly affected by concurrent administration of the buffered or enteric-coated formulations of didanosine. Therefore, amprenavir may be administered concurrently with either the buffered or the encapsulated enteric-coated bead formulation of didanosine in the fasting state. PMID- 12885097 TI - Impact of an alcohol withdrawal syndrome practice guideline on surgical patient outcomes. AB - STUDY OBJECTIVE: To standardize treatment of alcohol withdrawal syndrome (AWS) in surgical patients using an AWS practice guideline with a symptom-triggered approach. DESIGN: Prospective interventional (pilot group) and retrospective (comparison group). SETTING: University teaching hospital. PATIENTS: Thirty-eight trauma, orthopedic, and general surgery patients identified at risk for AWS in the pilot group, and 34 patients who were managed using nonstandardized approaches. INTERVENTIONS: At-risk patients in the pilot group were assessed using the AWS Type Indicator. They received lorazepam, clonidine, or haloperidol, based on AWS Type Indicator assessment and AWS practice guideline criteria. MEASUREMENTS AND MAIN RESULTS: A standardized symptom-triggered approach to managing AWS was expected to decrease the use of benzodiazepines, avoid undertreatment of adrenergic hyperactivity and delirium, decrease the need for sitters and physical restraints, and reduce hospital length of stay. Pilot patients received a mean of 23 mg less benzodiazepine (p=0.01), 0.1 mg more clonidine (p=0.01), and 20 mg less haloperidol (p=0.06) than comparison patients. Pilot patients also required significantly fewer sitter hours (p=0.04) and hours of restraint use (p=0.09) than comparison patients. No significant differences were found between groups for length of stay (p=0.77). CONCLUSIONS: This pilot project suggests that trauma, orthopedic, and general surgery patients at risk for AWS can be safely and effectively managed with a standardized, symptom triggered approach. Moreover, this approach decreased the amounts of benzodiazepines and haloperidol administered to patients at risk for AWS. PMID- 12885098 TI - Assessment of medication errors that involved drug allergies at a university hospital. AB - OBJECTIVES: To determine the reporting accuracy of true patient-related allergies to drugs in a large teaching institution (908 licensed beds), and to identify factors contributing to medication errors that involved drug allergies. Of particular interest was the accuracy of allergy information in the medical record and the occurrence of medication errors that involved penicillin antibiotics. METHODS: From a sample population of 340 patients, 50 adult patients admitted to our university-affiliated hospital who met criteria and had an allergy to selected drugs that was documented in the hospitalwide computer system were randomly selected and interviewed to determine the timing, nature, and extent of the reaction. Furthermore, data were collected from identified Medication Error Reports when an agent was prescribed for a patient with a reported allergy to that agent or class. Prospective data collection was conducted from November 2000 February 2001. Using the information obtained by the patient interview and chart documentation, we assessed the reported allergy. In addition, contributing factors for medication errors that involved drug allergies were identified. RESULTS: Of the sample population, 133 patients (39%) reported allergies to at least one drug. Allergies to beta-lactams, sulfonamides, and opioid narcotics were reported in 12.6% (43 patients), 9.1% (31), and 14.4% (49) of the sample population, respectively. Most agents involved in medication errors were beta lactam antibiotics, with an overwhelming number of these errors due to piperacillin-tazobactam (51.4%, 36 errors). Other drugs involved were ampicillin (10%, 7 errors), other beta-lactams (24.3%, 17 errors), opioid narcotics (10%, 7 errors), and sulfonamides (4.3%, 3 errors). Most contributing factors were classified as "MD [prescribing physician] not aware of allergy." CONCLUSION: These results suggest a need for ensuring that prescribers review each patient's allergy profile before order entry. PMID- 12885099 TI - Thiazolidinedione safety and efficacy in ambulatory patients receiving hemodialysis. AB - STUDY OBJECTIVES: To determine whether thiazolidinediones cause significant changes in intravascular volume, anemia, or chronic heart failure; to determine which thiazolidinedione, rosiglitazone or pioglitazone, has a greater propensity to cause these adverse effects; and to evaluate thiazolidinedione efficacy in patients with diabetes mellitus and end-stage renal disease who require hemodialysis. DESIGN: Retrospective chart review. SETTING: Ambulatory hemodialysis clinic. PATIENTS: Forty ambulatory patients receiving hemodialysis. MEASUREMENTS AND MAIN RESULTS: Of the 40 patients (26 men, 14 women, mean +/- SD age 64.8 +/- 11.5 yrs), diabetes mellitus was the cause of end-stage renal disease in 37 (92.5%). The men were older than the women (mean +/- SD age 67.65 +/- 11.43 yrs and 59.58 +/- 10.6 yrs, respectively, p=0.03). Additional demographic data collected were start date and cause of end-stage renal disease, comorbid conditions, drug profile, hospitalization dates, and reason for admission. Laboratory values were obtained for hematocrit, iron indexes (transferrin saturation and ferritin), mean corpuscular volume, and hemoglobin A1c (A1C); body weight before and after dialysis, and predialysis systolic and diastolic blood pressures were measured. All monitoring parameters were evaluated for 3 months before and after the start of therapy. Three patients were hospitalized for new or worsening chronic heart failure (two were receiving rosiglitazone therapy, one pioglitazone, p=0.555). Changes in A1C values were reviewed to determine thiazolidinedione efficacy; no statistical difference was observed between thiazolidinedione agents prescribed. Combined thiazolidinedione data yielded nonsignificant effects for all clinical and laboratory findings except A1C (-0.61%, p=0.05) and blood pressure (systolic -5.57 +/- 12.09 mm Hg, p=0.01; diastolic -3.24 +/- 6.17 mm Hg, p=0.002). CONCLUSION: Thiazolidinedione therapy is safe and effective for ambulatory patients receiving hemodialysis. However, as we found that these drugs reduced systolic and diastolic blood pressure, further investigation into this drug effect is warranted. PMID- 12885100 TI - Coadministration of milk thistle and indinavir in healthy subjects. AB - STUDY OBJECTIVE: To determine if milk thistle (silymarin) alters the pharmacokinetics of indinavir. DESIGN: Sequential crossover trial. SETTING: General clinical research center. SUBJECTS: Ten healthy subjects. INTERVENTION: Indinavir 800 mg 3 times/day was given for four doses on days 1 and 2. Silymarin 160 mg 3 times/day was given on days 3-15. On day 16 and for one dose on day 17, both drugs were given at the same dosages. MEASUREMENTS AND MAIN RESULTS: Indinavir's pharmacokinetic parameters were evaluated at steady state both before and after administration of 14 days of silymarin. Blood samples were collected 0.25, 0.5, 1, 2, 3, 4, and 5 hours after indinavir dosing and assayed by high performance liquid chromatography. The final pharmacokinetic model had first order absorption after a lag time, and two compartments with first-order elimination from the central compartment. When given alone and combined with silymarin, respectively, the geometric mean (95% confidence interval [CI]) steady state indinavir area under the plasma concentration-time curve was 20.7 hr x mg/L (15.3-28.2 hr x mg/L) and 19.4 hr x mg/L (15.8-23.6 hr x mg/L) and the trough plasma concentration was 0.340 mg/L (0.232-0.497 mg/L) and 0.232 mg/L (0.129 0.419 mg/L). CONCLUSION: Silymarin has no apparent effect on indinavir plasma concentrations. PMID- 12885101 TI - Statin-associated memory loss: analysis of 60 case reports and review of the literature. AB - OBJECTIVE: To review case reports of statin-associated memory loss as well as the available published evidence for and against such a link. METHODS: We searched the MedWatch drug surveillance system of the Food and Drug Administration (FDA) from November 1997-February 2002 for reports of statin-associated memory loss. We also reviewed the published literature (using MEDLINE) and prescribing information for these drugs. RESULTS: Of the 60 patients identified who had memory loss associated with statins, 36 received simvastatin, 23 atorvastatin, and 1 pravastatin. About 50% of the patients noted cognitive adverse effects within 2 months of therapy. Fourteen (56%) of 25 patients noted improvement when the statin was discontinued. Memory loss recurred in four patients who were rechallenged with the drug. None of the 60 reported cognitive test results. Two placebo-controlled trials found no benefits for statins on cognition or disability. One randomized controlled trial of simvastatin found no effects on cerebrospinal amyloid levels. In one small, randomized study, patients receiving statins showed a trend toward lower cognitive performance than those receiving placebo. Five observational studies found a lower risk of dementia among patients receiving statins. CONCLUSION: Current literature is conflicting with regard to the effects of statins on memory loss. Experimental studies support links between cholesterol intake and amyloid synthesis; observational studies indicate that patients receiving statins have a reduced risk of dementia. However, available prospective studies show no cognitive or antiamyloid benefits for any statin. In addition, case reports raise the possibility that statins, in rare cases, may be associated with cognitive impairment, though causality is not certain. PMID- 12885102 TI - Clinical relevance and management of drug-related QT interval prolongation. AB - Much attention recently has focused on drugs that prolong the QT interval, potentially leading to fatal cardiac dysrhythmias (e.g., torsade de pointes). We provide a detailed review of the published evidence that supports or does not support an association between drugs and their risk of QT prolongation. The mechanism of drug-induced QT prolongation is reviewed briefly, followed by an extensive evaluation of drugs associated with QT prolongation, torsade de pointes, or both. Drugs associated with QT prolongation are identified as having definite, probable, or proposed associations. The role of the clinician in the prevention and management of QT prolongation, drug-drug interactions that may occur with agents known to affect the QT interval, and the impact of this adverse effect on the regulatory process are addressed. PMID- 12885103 TI - Angiotensin-converting enzyme inhibitors and type 2 diabetic nephropathy: a meta analysis. AB - OBJECTIVE: To perform a meta-analysis on studies evaluating the effect of angiotensin-converting enzyme (ACE) inhibitors on diabetic nephropathy in patients with type 2 diabetes mellitus. METHODS: A computerized literature search was conducted for articles of studies comparing ACE inhibitors with a control in patients with diabetes, in which measurement of albuminuria or proteinuria was an outcome. Each article was abstracted by two of the authors. Data from the articles were presented as geometric or arithmetic means. The data were summarized separately by using standard techniques for meta-analysis. MAIN RESULTS: Statistically significant reductions in albuminuria were observed regardless of whether data were described with geometric or arithmetic means. Both were associated with significant heterogeneity. When studies reporting geometric means were stratified and analyzed, the heterogeneity was lost and statistically significant reductions in albuminuria were observed. The same procedure was repeated for studies reporting arithmetic means, but heterogeneity remained. CONCLUSION: The ACE inhibitors produce statistically significant reductions in albuminuria associated with significant heterogeneity of effect. Stratification reduces the heterogeneity and supports treatment with ACE inhibitors to reduce the progression of nephropathy in patients with type 2 diabetes mellitus. PMID- 12885104 TI - The role of multidrug efflux pumps in the antibiotic resistance of Pseudomonas aeruginosa and other gram-negative bacteria. Insights from the Society of Infectious Diseases Pharmacists. AB - Gram-negative bacteria remain clinically important pathogens in both hospital and community settings. Recent research indicates that efflux pumps play a prominent role in the multidrug resistance of Pseudomonas aeruginosa and many other gram negative bacteria. Four multidrug efflux pump systems have been well characterized in P. aeruginosa: MexA-MexB-OprM, MexC-MexD-OprJ, MexE-MexF-OprN, and MexX-MexY-OprM. These efflux pumps have different substrate specificities, and their production and activity can be increased by many factors commonly present in infections (e.g., high inocula of bacteria, low pH, and stationary phase growth). Moreover, fluoroquinolone antibiotics can commonly select mutants that constitutively overproduce Mex-Opr efflux pump systems. Based on most recent studies, the prevalence of efflux pump overproduction in clinical strains of P. aeruginosa may range from 14-75%. The best treatment for infections caused by bacteria that overproduce efflux pumps is unknown, but pharmacodynamic optimization of antibiotics and the use of antibiotic combinations that are substrates for different pump systems may represent reasonable strategies until more data are available. PMID- 12885105 TI - A novel method of estimating cost of therapy by using patient population characteristics: analysis of fluoroquinolones in various populations with different distributions of renal function. AB - INTRODUCTION: Formulary decisions regarding a given drug class are often made in the absence of patient outcome and/or sophisticated pharmacoeconomic data. Analyses that consider factors beyond simple acquisition costs may be useful in such situations. For example, the cost implications of using manufacturers' recommendations for dosing in patients with renal dysfunction may be important, depending on the distribution of various levels of renal function within a patient population. METHODS: Using four 1000-patient populations representing different renal function distributions and a fifth population of our medical center's distribution, we determined the costs of therapy for intravenous and oral levofloxacin, gatifloxacin, and moxifloxacin for a 10-day course of therapy for community-acquired pneumonia. Costs considered were average wholesale prices (AWPs), 50% of AWP, or same daily price, plus intravenous dose preparation and administration costs when applicable. Costs for each renal function distribution were examined for significant differences with an analysis-of-variance test. Also, costs of failing to adjust dosing regimens for decreased renal function were determined. RESULTS: Differences in fluoroquinolone costs (AWP, 50% AWP, or when matched as the same daily price) among the populations were found. When considering same daily prices, differences among populations ranged from about 35,000 dollars with intravenous gatifloxacin to more than 51,000 dollars for intravenous levofloxacin (all fluoroquinolones, p>0.05). Within a population, differences in costs among the intravenous fluoroquinolones ranged from 47,000 99,000 dollars. Rank orders of the drugs and population costs of therapy were affected by the pricing structure used and varied by the specific population and drug. Differences among the fluoroquinolones or populations were much smaller (<2100 dollars) when considering oral regimens. Costs potentially incurred by failing to adjust dosing for renal function were substantial. CONCLUSION: Formulary decisions can be facilitated by considering factors such as patient characteristics and related dosing in addition to simple acquisition costs. In our example, consideration of the distribution of renal function within a given patient population and related dosing for these fluoroquinolones revealed potentially important differences within the class. PMID- 12885106 TI - Propofol withdrawal syndrome in an adult patient with thermal injury. AB - A 48-year-old man with a history of ethanol abuse and bipolar disease fell asleep while smoking in an intoxicated state. The patient received a 30% total body surface area burn involving his face and upper torso that resulted in an inhalation injury. Several attempts at weaning from mechanical ventilation failed due to his extreme agitation, which was unresponsive to benzodiazepines, opiates, and antipsychotic agents. Propofol therapy was begun in combination with valproic acid, fluoxetine, and risperidone to assist in the treatment of his severe agitation associated with the bipolar disease, inhibiting ventilatory weaning. Repeated attempts to discontinue propofol were associated with withdrawal symptoms such as severe agitation, tremors, tachycardia, tachypnea, and hyperpyrexia. His symptoms resolved only after each time the propofol infusion was restarted. The patient received propofol for 95 days for management of his agitation before dying from refractory septic shock and multiple organ failure. PMID- 12885107 TI - Hypertriglyceridemia, acute pancreatitis, and diabetic ketoacidosis possibly associated with mirtazapine therapy: a case report. AB - A 44-year-old woman with a history of major depression and obsessive-compulsive disorder was prescribed mirtazapine. She came to the emergency department approximately 2 months after starting therapy; severe hypertriglyceridemia, acute pancreatitis, and diabetic ketoacidosis were diagnosed. Although these adverse effects have been reported in early clinical trials, we found only three published cases of subclinical pancreatitis possibly associated with mirtazapine therapy. We suspect that mirtazapine-associated hypertriglyceridemia had contributed to the development of acute pancreatitis and diabetic ketoacidosis in our patient. All these problems resolved with supportive care and discontinuation of mirtazapine. Her serum amylase, lipase, and lipid levels were normal 2 months after the acute event occurred. Health care providers should be aware of these possible adverse effects. Serum glucose and triglyceride levels should be measured at baseline and monitored regularly thereafter in all patients receiving mirtazapine therapy. PMID- 12885108 TI - Possible heart failure exacerbation associated with rosiglitazone: case report and literature review. AB - Increasing evidence suggests that neurohumoral manifestations of heart failure may lead to insulin resistance, predisposing patients with heart failure to the development of glucose intolerance or worsening of existing diabetes. Theoretically, insulin-sensitizing thiazolidinediones (TZDs) should be beneficial in this patient population. A 74-year-old man with well-compensated systolic dysfunction and longstanding type 2 diabetes mellitus treated with glyburide began therapy with rosiglitazone 4 mg/day, which was increased to 8 mg/day after 1 month. Two weeks later he was seen with a 5-kg weight gain, shortness of breath, bibasilar rales, +S3 gallop, and increased jugular venous distention. Twelve days later symptoms worsened, with pulmonary edema on chest radiograph, continued weight gain, and +4 pitting edema resistant to oral diuretics. The patient was admitted to the hospital for exacerbation of heart failure. Five days after discharge he was readmitted for similar symptoms, including an 11.8-kg weight gain. He reported adherence to drug therapy and diet. Rosiglitazone was immediately discontinued and 11 days later the man's weight stabilized to 79 kg and remained between 79 and 80 kg 2 and 3 months after discharge. This case demonstrates that TZDs may precipitate weight gain and pulmonary and peripheral edema in patients with stable heart failure. Earlier reports documented similar symptoms in patients without a history of heart failure. Although current recommendations state that TZDs should not be administered to patients with New York Heart Association class III or IV disease, practitioners should be aware that these adverse effects also may occur in patients with milder forms heart failure as well as those without heart failure. PMID- 12885109 TI - Temporal construal. AB - Construal level theory proposes that temporal distance changes people's responses to future events by changing the way people mentally represent those events. The greater the temporal distance, the more likely are events to be represented in terms of a few abstract features that convey the perceived essence of the events (high-level construals) rather than in terms of more concrete and incidental details of the events (low-level construals). The informational and evaluative implications of high-level construals, compared with those of low-level construals, should therefore have more impact on responses to distant-future events than near-future events. This article explores the implications of construal level theory for temporal changes in evaluation, prediction, and choice. The authors suggest that construal level underlies a broad range of evaluative and behavioral consequences of psychological distance from events. PMID- 12885110 TI - Driven by information: a tectonic theory of Stroop effects. AB - The goal of avoiding distraction (e.g., ignoring words when naming their print colors in a Stroop task) is opposed intrinsically by the penchant to process conspicuous and correlated characteristics of the environment (e.g., noticing trial-to-trial associations between the colors and the words). To reconcile these opposing forces, the authors propose a tectonic theory of selective attention in which 2 memory-based structures--dimensional imbalance and dimensional uncertainty--drive selection by processing salient, surprising, and/or correlated information contained within and across stimulus dimensions. Each structure modulates the buildup of excitation to targets and the buildup of inhibition to distractors and to memories of previous stimuli. Tectonic theory is implemented to simulate the impact of 4 types of context on the presence, magnitude, and direction of congruity effects and task effects in the Stroop paradigm. The tectonic model is shown to surpass other formal models in explaining the range and diversity of Sroop effects. PMID- 12885111 TI - Comparison processes in social judgment: mechanisms and consequences. AB - This article proposes an informational perspective on comparison consequences in social judgment. It is argued that to understand the variable consequences of comparison, one has to examine what target knowledge is activated during the comparison process. These informational underpinnings are conceptualized in a selective accessibility model that distinguishes 2 fundamental comparison processes. Similarity testing selectively makes accessible knowledge indicating target-standard similarity, whereas dissimilarity testing selectively makes accessible knowledge indicating target-standard dissimilarity. These respective subsets of target knowledge build the basis for subsequent target evaluations, so that similarity testing typically leads to assimilation whereas dissimilarity testing typically leads to contrast. The model is proposed as a unifying conceptual framework that integrates diverse findings on comparison consequences in social judgment. PMID- 12885112 TI - Meaning through syntax: language comprehension and the reduced relative clause construction. AB - A new explanation is proposed for a long standing question in psycholinguistics: Why are some reduced relative clauses so difficult to comprehend? It is proposed that the meanings of some verbs like race are incompatible with the meaning of the reduced relative clause and that this incompatibility makes sentences like The horse raced past the barn fell unacceptable. In support of their hypotheses, the authors show that reduced relatives of The horse raced past the barn fell type occur in naturally produced sentences with a near-zero probability, whereas reduced relatives with other verbs occur with a probability of about 1 in 20. The authors also support the hypotheses with a number of psycholinguistic experiments and corpus studies. PMID- 12885113 TI - Hypothesis testing and theory evaluation at the boundaries: surprising insights from Bayes's theorem. AB - Because the probability of obtaining an experimental finding given that the null hypothesis is true [p(F?H0)] is not the same as the probability that the null hypothesis is true given a finding [p(H0?F)], calculating the former probability does not justify conclusions about the latter one. As the standard null hypothesis significance-testing procedure does just that, it is logically invalid (J. Cohen, 1994). Theoretically, Bayes's theorem yields p(H0?F), but in practice, researchers rarely know the correct values for 2 of the variables in the theorem. Nevertheless, by considering a wide range of possible values for the unknown variables, it is possible to calculate a range of theoretical values for p(H0?F) and to draw conclusions about both hypothesis testing and theory evaluation. PMID- 12885114 TI - A recurrent connectionist model of group biases. AB - Major biases and stereotypes in group judgments are reviewed and modeled from a recurrent connectionist perspective. These biases are in the areas of group impression formation (illusory correlation), group differentiation (accentuation), stereotype change (dispersed vs. concentrated distribution of inconsistent information), and group homogeneity. All these phenomena are illustrated with well-known experiments, and simulated with an autoassociative network architecture with linear activation update and delta learning algorithm for adjusting the connection weights. All the biases were successfully reproduced in the simulations. The discussion centers on how the particular simulation specifications compare with other models of group biases and how they may be used to develop novel hypotheses for testing the connectionist modeling approach and, more generally, for improving theorizing in the field of social biases and stereotype change. PMID- 12885115 TI - Egocentrism versus protocentrism: the status of self in social prediction. AB - In this article, the author discusses the limitations of the egocentric view of self in which self serves as an automatic filter, inhibiting access to alternative representations of others' thoughts and feelings. The author then outlines a protocentric model, the self-as-distinct (SAD) model, in which generic representations of prototypic others serve as the default; representations of self, specific others, or categories encode only distinctiveness from generic knowledge about prototypic others. Thus, self-knowledge is distributed both in generic representations in which self and prototypic others are undifferentiated and in a self-representation that encodes distinctiveness. The self representation does not serve to make predictions about others because it encodes how self differs from the generic representation of others. Predictions that are the same about self and others are protocentric, based on generic knowledge that serves as the default. The SAD model parsimoniously accounts for many inconsistent findings across various domains in social cognition. PMID- 12885116 TI - When egocentrism breeds distinctness--comparison processes in social prediction: comment on Karniol (2003). AB - In contrast with R. Karniol's (2003) protocentric model, this article proposes an egocentric comparison model of social prediction. To predict the experiences and reactions of a target other, judges are assumed to relate self-knowledge to the target via a process of comparing the target with the self. Two alternative comparison processes are distinguished. Similarity testing yields target judgments that are consistent with the self, whereas dissimilarity testing leads to target judgments that are inconsistent with the self. From this perspective, social prediction is egocentric in that it makes use of self-knowledge as its primary representational basis. At the same time, egocentric prediction does not require the self to be seen and judged as similar to the target other. Conceptualizing social predictions as social comparison suggests that predicting the experiences and reactions of others is a highly flexible process. PMID- 12885117 TI - Return of the ego--self-referent information as a filter for social prediction: comment on Karniol (2003). AB - The protocentrism paradigm of social prediction (R. Karniol, 2003) challenges the egocentrism paradigm tacitly accepted by many researchers. The author reviews the 2 paradigms comparatively by focusing on 3 conceptual and 3 empirical issues. On conceptual grounds, the author suggests that the egocentrism paradigm has been proven useful because of (a) its greater breadth and parsimony, (b) the difficulties in documenting the origin of protocenters, and (c) the indeterminate nature of self-as-distinct tags (which are crucial to protocentrism). On empirical grounds, the author argues that in research on perceptions of self other similarities, the egocentric process of social projection is well established. Self-referent knowledge (a) is most readily accessible, (b) receives greater weight in prediction tasks than does other-referent knowledge, and (c) tends to be suppressed only temporarily, with effort, and incompletely. PMID- 12885118 TI - On the status of self in social prediction: comment on Karniol (2003). AB - A challenge to mainstream notions on the status of the self in social prediction is welcome. The self-as-distinct model (R. Karniol, 2003) is thoughtful, provocative, and parsimonious, but it is also underspecified, undertested, and selective in its treatment of the evidence. More important, the model does not provide compelling answers to issues pertaining to the origins of prototypic social knowledge, the status of self-knowledge, the content of the self representation, whether the use of self in social prediction is a logical contradiction, and whether the self's role in social prediction is amotivated. PMID- 12885119 TI - Synthesis and properties of phosphorylated 3'-O-beta-D-ribofuranosyl-2' deoxythymidine. AB - A strategy was developed for the synthesis of 3'-O-beta-D-ribofuranosyl 2' deoxythymidine derivatives using three different protecting groups, which allows the synthesis of a phosphoramidite building block for oligonucleotide synthesis. Likewise the 5'-O- and 5''-O-phosphorylated analogues were synthesized and their conformation was determined using NMR spectroscopy. PMID- 12885120 TI - Synthesis, structural studies, and biological evaluation of some purine substituted 1-aminocyclopropane-1-carboxylic acids and 1-amino-1 hydroxymethylcyclopropanes. AB - The novel purine derivatives of 1-aminocyclopropane-1-carboxylic acid (8 and 9) and 1-amino-1-hydroxymethylcyclopropane (12 and 13) with methylene spacer between the base and the cyclopropane ring were prepared by multistep synthetic route involving alkylation of adenine and 6-(N-pyrrolyl)purine with 2-hydroxy-methyl-1 aminocyclopropane-1-carboxylic acid derivative 3 as a key reaction. All novel compounds were racemic. The N-9 substitution of the purine ring and the Z configuration of the cyclopropane ring in 4-13 were deduced from their 1H and 13C NMR spectra by analyses of chemical shifts, H-H coupling constants and connectivities in two-dimensional homo- and heteronuclear correlation spectra. An unequivocal proof of the stereostructure of 1, 4 and 5 was obtained by their X ray structure analysis. The novel compounds were evaluated on cytostatic and antiviral activities in several cell lines. The 6-(N-pyrrolyl)purine derivative of 1,2-aminocyclopropane alcohol 12 exhibited a more pronounced inhibitory activity against the proliferation of cervical carcinoma (HeLa) and human fibroblast (WI-38) cells than other types of tumor cell lines. None of the compounds showed inhibitory activities against cytomegalovirus, varicella-zoster virus or other viruses. PMID- 12885121 TI - C-5 modified nucleosides: direct insertion of alkynyl-thio functionality in pyrimidines. AB - A route is presented to append, in a single step, alkynyl thioesters to the 5 position of a pyrimidine ring of a nucleoside that is unprotected. These products should be useful to support in vitro selection experiments with functionalized DNA. PMID- 12885122 TI - The analogue of thymidine triphosphate containing a methylene group in place of the 5' oxygen can serve as a substrate for reverse transcriptase. AB - The thymidine 5'-triphosphate analogue containing a methylene group in place of the 5' oxygen atom can be prepared using modifications of published procedures and can substitute for the natural thymidine triphosphate in chain extension reactions catalyzed by Moloney-MLV reverse transcriptase. Using rabbit beta globin mRNA as the template together with an appropriate primer, the enzyme readily makes full-length DNA transcripts in which all thymidine 5' oxygen atoms have been replaced with methylene groups. In sequence analyses using the partial depurination procedure, the analogue DNA transcript produces electrophoretic gel patterns identical with those of the corresponding natural DNA transcript. Experiments on second strand synthesis using the four regular triphosphates show that the analogue DNA transcript, like the natural transcript, can serve as a template. The two DNA duplexes (natural/natural and analogue/natural) formed by these reactions produce similar electrophoretic cleavage patterns when treated with either of the endonucleases HaeIII and EcoRI. However, further studies on template properties indicate that, while the enzyme makes a full-length product when using the analogue substrate with a natural DNA strand as template, it appears unable to use the analogue transcript as template with the analogue triphosphate as substrate during second strand synthesis. Preliminary experiments have also been carried out with a DNA polymerase. No products are detected reactions using Taq polymerase with PCR protocols containing the analogue triphosphate as the only source of thymidine. PMID- 12885123 TI - Synthesis of triazole nucleoside derivatives. AB - 5'-O-Mesyl-2',3'-O-isopropylidene ribonucleosides (4 and 12) were converted to their 5'-substituted nucleosides in good yields by reacted with NaN3 or KI. 2',3' O-Isopropylidene ribonucleosides (3 and 11) were prepared in good yields from ribonucleosides 1 and 2 with a reaction mixture of acetone and triethyl orthoformate instead of using acetone diethyl acetal. Compound 1 or 2 was treated with 2-acetoxyisobutyryl halide (Cl or Br) to give 1-[2-O-acetyl-3-halo-3-deoxy-5 O-(2,5,5-trimethyl-1,3-dioxolan-4-on-2-yl)-beta-D-xylofuranosyl]-1,2,4-triazole-3 carboxamide (19, 22, and 23) in high yields. Instead of using 2-acetoxyisobutyryl bromide, the mixture of 2-acetoxyisobutyryl chloride and NaBr was employed in the synthesis of 22 and 23. Treatment of 19 with an activated Zn/Cu couple and deprotection gave 2',3'-anhydro nucleoside (21), and treatment of 22 and 23 with an activated Zn/Cu couple and a little of HOAc and deprotection gave corresponding 2',3'-unsaturated triazole nucleosides (24 and 25), respectively. The biological activity of the compounds (7-10, 15-18, and 24) was examined in human liver cancer cells (A-549), lung cancer cells (BEL-7402), and Flu-A cells. PMID- 12885124 TI - Synthesis of carbocyclic 1-[4-(hydroxymethyl)cyclopent-2-enyl]-1,2,4-triazole-3 carboxamide and its derivatives. AB - The synthesis of carbocyclic 1-[4-(hydroxymethyl)cyclopent-2-enyl]-1,2,4-triazole 3-carboxamide (6a) and its derivatives was achieved from triol 10 in excellent overall yield. This route involves a Pd(0)-catalyzed coupling reaction as a key step. PMID- 12885125 TI - 2-Methyl-5-tert-butylthiophenol--an odorless deprotecting reagent useful in synthesis of oligonucleotides and their analogs. AB - A solution of 2-methyl-5-tert-butylthiophenol and triethylamine in acetonitrile efficiently removed the methyl protecting groups from phosphate and phosphorothioate triesters and yielded oligonucleotides of high quality. This inexpensive odorless liquid is not toxic and is a suitable replacement for hazardous thiophenol and other reagents often used for this purpose. PMID- 12885126 TI - Solid phase synthesis of phosphorothioate oligonucleotides utilizing diethyldithiocarbonate disulfide (DDD) as an efficient sulfur transfer reagent. AB - Diethyldithiodicarbonate (DDD), a cheap and easily prepared compound, is found to be a rapid and efficient sulfurizing reagent in solid phase synthesis of phosphorothioate oligodeoxyribonucleotides via the phosphoramidite approach. Product yield and quality based on IP-LC-MS compares well with high quality oligonucleotides synthesized using phenylacetyl disulfide (PADS) which is being used for manufacture of our antisense drugs. PMID- 12885127 TI - 6-(Levulinyloxymethyl)-3-methoxy-2-nitrobenzoyl and 2-(levulinyloxymethyl)-5 methoxy-4-nitrobenzoyl groups as novel base-labile groups for 5'-hydroxy protection in solid-phase oligonucleotide synthesis. AB - The 6-(levulinyloxymethyl)-3-methoxy-2-nitrobenzoyl (LMMoNBz) and 2 (levulinyloxymethyl)-5-methoxy-4-nitrobenzoyl (LMMpNBz) groups were developed as novel base-labile groups for 5'-hydroxy protection in solid-phase oligonucleotide synthesis. A comparative study of the utility of LMMoNBz, LMMpNBz, and 2 (levulinyloxymethyl)-5-nitrobenzoyl (LMNBz) groups is described. PMID- 12885128 TI - A short review of transmissible spongiform encephalopathies, and guidelines for managing risks associated with chronic wasting disease in captive cervids in zoos. AB - The transmissible spongiform encephalopathies (TSEs) represent an emerging group of diseases that have been labeled as "prion diseases" because of the recent characterization of the infectious agent. TSEs are caused by prions, which induce neurodegenerative fatal diseases in humans and animals. Some TSEs (scrapie and kuru), have existed in both animals and humans for a very long time, whereas others such as bovine spongiform encephalopathy and variant Creutzfeld-Jakob disease have either recently emerged or are more thoroughly described and recognized. It is obvious that the medical community will be forced to consider these diseases in humans and animals for the future. This article offers a short review of the TSEs of immediate concern to zoo and wildlife veterinarians and wildlife biologists and suggests risk management strategies for the prevention of these diseases, with special focus on chronic wasting disease of cervids in North America. PMID- 12885129 TI - A review of alpha2 adrenoreceptor agonists and the development of hypoxemia in domestic and wild ruminants. AB - Alpha2 agonists are commonly used in combination with other anesthetic agents to chemically immobilize wildlife and may cause potentially serious hypoxemia in wild ruminants. In some domestic ruminants, they are associated with significant changes to the pulmonary parenchyma, increased venous admixture, pulmonary edema, and hypoxemia. Effective and safe immobilization of these animals requires knowledge of the mechanisms behind these changes and the methods that offset their effects, including the use of supplemental oxygen. PMID- 12885130 TI - Identification of Haemobartonella felis (Mycoplasma haemofelis) in captive nondomestic cats. AB - This study was undertaken to determine whether Haemobartonella felis (Mycoplasma haemofelis), the causative bacterial agent of feline infectious anemia, infects nondomestic cats. Routine complete blood count and polymerase chain reaction (PCR) were performed to detect the gene for 16S ribosomal RNA for the organism. Sixty-four blood samples were collected from 54 nondomestic cats, including tigers (Panthera tigris), cheetahs (Acinonyx jubatus), lions (P. leo), mountain lions (Felis concolor), snow leopards (P. unica), and a jaguar (P. onca). Some cats were sampled on two or three different dates. Two tigers were positive for H. felis by PCR analysis. As previously described in domestic cats, the parasitemia appears to be intermittent in nondomestic cats. PMID- 12885131 TI - Ultrasonographic imaging of the sandhill crane (Grus canadensis) intertarsal joint. AB - Tendon ossification in the crus and tarsometatarsal regions of cranes makes ultrasonography difficult everywhere except for joints, where ossification is absent. Normal ultrasonographic anatomy of the adult Florida sandhill crane (Grus canadensis pratensis) intertarsal joint is described on the basis of ultrasonography that was performed on the limbs of a cadaver, which were dissected and cross-sectioned subsequently, to correlate ultrasonographic images with anatomic structures. Intertarsal joints of five normal sandhill cranes and two cranes with known intertarsal abnormalities were then imaged bilaterally in transverse and sagittal planes. Ultrasonographic imaging can be used to evaluate the soft-tissue structures on the dorsal and plantar aspects of the intertarsal joint of cranes, and it is a useful adjunct to physical and radiographic examination for localizing injuries to this area. PMID- 12885132 TI - Evaluation of propofol and medetomidine-ketamine for short-term immobilization of Gulf of Mexico sturgeon (Acipenser oxyrinchus de soti). AB - Parenteral anesthetic protocols for short-term immobilization were evaluated in twenty 4-yr-old Gulf of Mexico sturgeon (Acipenser oxyrinchus de soti). An initial dose-response trial determined the efficacy of either propofol (3.5-7.5 mg/kg. i.v.) or combinations of medetomidine (0.03-0.07 mg/kg, i.m.)-ketamine (3 7 mg/kg, i.m.). A subsequent study evaluated the physiologic effects of propofol (6.5 mg/kg, i.v.)-induced anesthesia and anesthesia induced with a medetomidine (0.06 mg/kg, i.m.)-ketamine (6 mg/kg i.m.) combination. The effects of medetomidine were reversed at 30 min with atipamezole (0.30 mg/kg, i.m.). Both drug protocols provided adequate short-term immobilization for minor diagnostic procedures. Sturgeon receiving propofol were in a light plane of anesthesia within 5 min after drug administration, whereas only 30% of the medetomidine ketamine group reached a light plane of anesthesia in the same time period. Both propofol and medetomidine-ketamine resulted in mild bradycardia and apparent respiratory depression, with propofol producing more profound effects. At the dosages used in this study, both propofol and the medetomidine-ketamine combination effectively induced a light plane of anesthesia. Induction times were shorter in the propofol group. PMID- 12885133 TI - Hematologic and serum biochemistry values in two species of free-ranging porcupines (Coendou prehensilis, Coendou melanurus) in French Guiana. AB - Individuals of two porcupine species (Coendou prehensilis and Coendou melanurus) were translocated during the flooding of a forest at a hydroelectric dam site in French Guiana. Blood samples were collected for 11 mo to determine hematologic and serum chemistry reference mean values and ranges and to look for blood parasites. Male C. prehensilis had significantly higher hemoglobin levels, packed cell volumes, and creatinine and potassium values than did females of the same species. Coendou prehensilis had significantly lower amylase levels but higher creatinine, calcium, lactate dehydrogenase, and gamma glutamyl transferase levels than C. melanurus. PMID- 12885134 TI - Comparison of the efficacy and cardiorespiratory effects of medetomidine-based anesthetic protocols in ring-tailed lemurs (Lemur catta). AB - The relative efficacies and cardiorespiratory effects of three injectable anesthetic combinations containing medetomidine were evaluated in ring-tailed lemurs (Lemur catta). In addition, the direct effects of medetomidine on heart rate and blood pressure were evaluated in lemurs anesthetized with isoflurane. For injectable anesthesia, captive adult ring-tailed lemurs were anesthetized with medetomidine and ketamine (0.04-0.06 mg/kg, i.m. and 3 mg/kg, i.m., respectively), medetomidine, butorphanol, and ketamine (0.04 mg/kg, i.m., 0.4 mg/kg, i.m., and 3 mg/kg, i.m., respectively), or medetomidine, butorphanol, and midazolam (0.04 mg/kg, i.m., 0.4 mg/kg, i.m., and 0.3 mg/kg, i.m., respectively). For inhalation anesthesia, lemurs were mask-induced and maintained with isoflurane for 30 min before receiving medetomidine (0.04 mg/kg, i.m.). Sedation produced by medetomidine-ketamine was unpredictable and of short duration. Both medetomidine-butorphanol-ketamine (MBK) and medetomidine-butorphanol-midazolam (MBMz) provided adequate anesthesia for routine physical exams; however, the effects of MBMz lasted longer than those of MBK. Heart rates and respiratory rates were within clinically normal ranges for all groups, and lemurs remained normotensive throughout the study. Common side effects such as hypertension and bradycardia associated with the use of alpha2-adrenergic receptor agonist combinations in other species were not observed. Likewise, medetomidine administration had no effect on HR in lemurs receiving isoflurane. Lemurs in all groups were well ventilated and remained well oxygenated throughout the procedures, though arterial partial pressure of O2 was lowest in the MBMz group. All three injectable medetomidine combinations were effective in ring-tailed lemurs but only MBK and MBMz provided adequate depth and duration of anesthesia for use as sole regimes. For many clinical procedures in lemurs, MBMz offers advantages over MBK because of its longer duration of action and its rapid and more complete reversibility with specific antagonists. PMID- 12885135 TI - Steady-state plasma concentrations of itraconazole after oral administration in Kemp's ridley sea turtles, Lepidochelys kempi. AB - Pharmacokinetic studies of antifungal agents in reptiles are uncommon. Itraconazole, which has been used prophylactically in juvenile sea turtles suffering from hypothermia (cold stunning) on a regular basis, was evaluated for steady-state plasma concentrations. Five Kemp's ridley sea turtles (Lepidochelys kempi) receiving itraconazole at several dosages in a rehabilitation program had blood collected within 24 hr to estimate dosing frequency. Subsequently, serial blood samples of Kemp's ridley sea turtles that were given itraconazole at several dosages for 30 days to treat cold stunning were collected at various intervals to evaluate itraconazole plasma concentrations. Tissue samples were collected from one Kemp's ridley that died during rehabilitation. Plasma concentrations of itraconazole (and of hydroxyitraconazole [OH-ITRA], one of its major bioactive metabolites) were determined using a modified, validated reverse phase high-performance liquid chromatography technique. Itraconazole concentrations in tissues were determined by bioassay to be far greater than the plasma concentrations measured in any of the turtles. At a 15-mg/kg dosage, the half-life (t1/2) was 75 hr for itraconazole and 55 hr for OH-ITRA. All dosages produced adequate concentrations in some turtles, but consistent therapeutic concentrations were produced only at 15 mg/kg q72hr and 5 mg/kg s.i.d., with the latter producing the highest plasma concentrations. PMID- 12885136 TI - Cutaneous fibropapilloma in a mountain lion (Felis concolor). AB - A 12-yr-old mountain lion (Felis concolor) developed a 0.5-cm3 raised nonpigmented and nonulcerated mass between the lip and the nasal planum. The tumor was surgically removed and diagnosed histologically as a fibropapilloma. The tumor recurred 1 yr later, at which time it was again excised, and the diagnosis was reconfirmed by biopsy. Frozen tissue from the second excision was submitted for polymerase chain reaction testing for papillomavirus. The 176-base pair polymerase chain reaction product recovered from the tumor was cloned and sequenced. The papillomavirus had 96% homology with a papillomavirus previously retrieved from a fibropapilloma in a domestic cat and is the next most closely related to bovine papillomavirus type 1. This is the first report of a virus associated fibropapilloma in a mountain lion. PMID- 12885137 TI - Mycobacteriosis in two captive Florida manatees (Trichechus manatus latirostris). AB - Two male Florida manatees (Trichechus manatus latirostris) died at the marine aquarium in Inagi City, Tokyo, Japan. Acid-fast bacteria were demonstrated in tuberculoid nodules in the lungs from both manatees. Mycobacterium marinum and M. fortuitum were isolated from one manatee; M. marinum and M. kansasii were cultured from the second animal. This report confirms the pathogenicity and potentially fatal outcome of mycobacterial infection in manatees. In addition, the pathologic response to infection with these mycobacteria in manatees is similar to that associated with Mycobacterium spp. in other animals. PMID- 12885138 TI - Detection of Toxoplasma gondii IgG antibodies in juvenile great grey kangaroos, Macropus giganteus giganteus. AB - Serial IgG antibody titers to Toxoplasma gondii from three juvenile great grey kangaroos (Macropus giganteus giganteus) with clinical signs consistent with toxoplasmosis were compared with titers from three clinically normal juveniles of similar age. The modified agglutination test was used to measure antibodies to T. gondii. Results demonstrated declining antibody titers to T. gondii in five of six animals. Antibody titers were similar in animals with and without clinical signs. This was interpreted as an indication that maternal IgG antibodies to T. gondii were transferred to juveniles, presumably in milk, rather than an indication of clinical disease. Therefore, clinicians should interpret Toxoplasma serology results in juvenile macropods cautiously. PMID- 12885139 TI - Intraocular nematodiasis caused by Parelaphostrongylus tenuis in an eland antelope (Taurotragus oryx). AB - A 10-mo-old female eland (Taurotragus oryx) at the Wilds exhibited recalcitrant, progressive unilateral uveitis for a 5-wk period, despite constant medical treatment. Unilateral enucleation was performed because of blindness and animal discomfort evidenced by continuous blepharospasm. Histopathologic examination of the eye demonstrated intraocular larvae morphologically consistent with Parelaphostrongylus tenuis, the first known case of intraocular P. tenuis migration. This animal subsequently was euthanatized because of severe, nonresponsive neurologic signs associated with P. tenuis infection. PMID- 12885140 TI - Eurasian otter (Lutra lutra), a definitive host for Dirofilaria immitis. AB - A mature male and a mature female Dirofilaria immitis were found in the right ventricle of the heart of a naturally infected 2-yr-old male Eurasian otter (Lutra lutra) that had died of severe lung congestion at a zoo in South Korea. Both developing embryos and microfilariae were present in the uterus of the female D. immitis. Although circulating microfilariae were not detected in blood or tissue, the Eurasian otter may serve as a definitive host for D. immitis. PMID- 12885141 TI - Immobilization of sun bears (Helarctos malayanus) with medetomidine-zolazepam tiletamine. AB - A mixture of medetomidine (50.0 microg/kg, i.m.) and zolazepam-tiletamine (2.0 mg/kg, i.m.) effectively immobilized 16 sun bears (Helarctos malayanus) for more than 1 hr with good myorelaxation and minimal effects on cardiorespiratory performance during 22 immobilizations. All bears were immobilized once, except for six individuals that were immobilized twice. Atipamezole (250 microg/kg, i.v.) effectively reversed medetomidine-induced sedation and reduced recovery time significantly. Respiratory rates of immobilized bears did not change significantly over time. Rectal body temperature and heart rate decreased significantly after 10 min of immobilization. Hematologic and serum biochemical parameters did not change significantly within 30 min of induction. PMID- 12885142 TI - Use of blood culture as a nonlethal method for isolating bacteria from fish. AB - Simple nonlethal blood culture methodology, an alternative to euthanasia for diagnosing systemic bacterial infections in fish, is described. Blood was extracted from the caudal vein of 20 individuals of five fish species, incubated in brain-heart infusion broth, and then plated onto enriched blood agar. Nine of these fish were subsequently euthanized and necropsied for confirmatory tissue cultures. Five species of bacteria were isolated from the blood cultures from nine fish, and the tissue culture results in euthanized, necropsied fish agreed with the blood culture results in all cases. All the fish that were not euthanized survived for 24 hr, although two heavily parasitized fish subsequently died. PMID- 12885143 TI - Subdimensions and subscales in measurement and instrument development. PMID- 12885144 TI - Evaluation of the diabetes self care scale: an illustration of the Rasch model of measurement. AB - This pilot study illustrates the use of a Rasch measurement model to evaluate the psychometric properties of the Diabetes Self Care Scale (DSCS). A convenience sample of 50 adults with diabetes at two local hospitals (one for profit and one public) in southern Louisiana responded to the survey questions. The instrument is comprised of 27 items and 6 rating categories. The Bigsteps Rasch measurement software was used for all analyses. Results indicated that respondent separation reliability was low (0.63), but item separation reliability was satisfactory (0.91). Internal validity was supported. Twenty-six items measure a single construct of diabetes self care. Construct validity was supported by the meaningfulness of the item difficulty order and by the consistency of that order across respondents. Rating scale analysis revealed that middle categories were not distinct steps in the progression of increasing diabetes self care. In addition to deleting an item producing inconsistent responses and combining adjacent categories (categories 1 and 2, and categories 3, 4, and 5), four recommendations for future research are provided. PMID- 12885145 TI - An introduction to Rasch measurement for scale development and person assessment. AB - This article: a) outlines some of the limitations of True Score Theory, b) discusses some advantages of Rasch measurement, and c) compares the results of Rasch and factor analytic techniques applied to a common data set. Using a mail survey, 206 female members of a national nursing association completed Ryckman's (1982) Perceived Physical Ability Scale. Exploratory factor analysis identified one factor with a Cronbach alpha of .78. Rasch fit statistics also identified one dimension with a person reliability of .76. Rasch methods were also used to investigate the rating scale functioning, additional construct validity issues, and person diagnosis. Discrepancies between the two estimates of internal consistency are discussed and differences in the utility of the two methodologies are reviewed. PMID- 12885146 TI - Developing and testing of the Foot Care Confidence Scale. AB - This study developed and tested the Foot Care Confidence Scale (FCCS), an instrument designed to measure the confidence (self-efficacy) diabetics have in performing foot self care. Content validity was assessed using Lynn's (1986) Content Validity Index (CVI) with a total of 100% content validity (Sloan, 1993). The FCCS was administered to four persons with diabetes (Sloan, 1994) and pilot tested with 41 healthy older adults (Wills & Sloan, 1995). Based on the feedback from the pilot testing and CVI panel of judges, as well as additional literature review, the instrument was revised. A study involving 122 community-dwelling older adults with a diagnosis of diabetes for five or more years was conducted. The FCCS had a Cronbach's alpha of 0.92. Factor analysis revealed that all 12 items loaded on one factor and all 12 items are needed to measure confidence to care for one's feet. The FCCS is a practical instrument for use in many settings, especially where time constraints exist. It may assist the nurse in evaluating and improving outcomes of self care of the feet of persons with diabetes mellitus. PMID- 12885147 TI - The Friedrich-Lively Instrument to Assess the Impact of Schizophrenia on Siblings (FLIISS): Part I--instrument construction. AB - Siblings of persons with schizophrenia may provide primary or secondary care for their sibling and support to parents who are primary care givers. These siblings experience stress and the accompanying sequelae of decreased quality of life, grief, chronic illness, and symptoms of depression and anxiety. Comprehensive measures of sibling stress have not been published. Before interventions to help siblings can be created, health professionals need such an assessment. The Friedrich-Lively Instrument to Assess the Impact of Schizophrenia on Siblings (FLIISS) has been developed to meet the needs of both health workers and siblings. The instrument has a strong conceptual basis adapted from Pearlin's model of stress in caregiving and was preceded by pilot work that evaluated both quantitative and qualitative data. In this article, (Part I), the development of the instrument and its relationships to the conceptual model are described. Health professionals can use the instrument for collecting information that will increase their ability to identify sources of stress faced by siblings of persons with schizophrenia, and from this assessment they may develop interventions for this underserved population. PMID- 12885148 TI - The Friedrich-Lively Instrument to Assess the Impact of Schizophrenia on Siblings (FLIISS): Part II--reliability and validity assessment. AB - Siblings of persons with schizophrenia who provide primary or secondary care for their sibling and support to parents may experience stress, decreased quality of life, grief, chronic illness, depression, and anxiety. The effects on siblings have not been widely studied and there have not been appropriate instruments to measure the types and degree of stress and associated mental, emotional, and physical distress. The Friedrich-Lively Instrument to Assess the Impact of Schizophrenia on Siblings (FLIISS) is a comprehensive instrument based on extensive research and work with siblings of persons with schizophrenia. Construction of the FLIISS and its relationship to a conceptual model of stress is presented in an article (Part I) elsewhere in this issue. In this article (Part II), reliability results and construct validity are reported for the FLIISS. The FLIISS demonstrates appropriate usefulness and content, acceptable internal consistency, and strong construct validity. Health professionals can use the instrument for collecting information that will increase their ability to identify sources of stress faced by siblings of persons with schizophrenia and develop interventions to help these siblings. PMID- 12885149 TI - Development and testing of the Labor Support Scale. AB - Nurses use interventions and supportive strategies to help mothers throughout labor, yet little research examines intrapartum nurses' labor support techniques. The purpose of the study was to develop and test a self-report instrument, the Labor Support Scale (LSS), to describe the frequency with which nurses perform interventions and nurses' perceptions of the helpfulness of interventions. Steps for instrument development were item generation, content validity testing, piloting, refining, and administering the questionnaire in two studies (n = 307, n = 472). Internal consistency reliability was .90 and .92 for the frequency and helpfulness portions of the instrument (respectively). Exploratory factor analysis, known groups technique, content analysis, and discriminant analysis evaluated validity. In both phases, instrument psychometrics provided evidence of content, construct, and discriminant validity. PMID- 12885150 TI - Use of the Edmonton Research Orientation Scale with nurses. AB - Improved understanding of the determinants of research utilization is fundamental to developing and testing strategies to increase research utilization. Inconsistent findings in this field of research about research utilization may be attributable, in part, to lack of development of measures. This research tested the internal consistency and construct validity of the Edmonton Research Orientation Scale (EROS) and its four subscales (Valuing Research, Research Involvement, Being on the Leading Edge, and Evidence-Based Practice), which are promising measures of research utilization and attitudes towards research. One hundred eighty-five registered nurses in a pediatric teaching hospital completed the EROS. Nurses who reported higher levels of education, better understanding of research topics or participation in quality management or research projects also reported higher levels on the EROS and EROS subscales. The Valuing Research and Evidence-Based Practice subscales were associated with having taken courses in research design and statistics. The findings suggest that the EROS and the Valuing Research and Evidence-Based Practice subscales may be used to measure nurses' attitudes towards research and research utilization. PMID- 12885151 TI - The search for a practical approach to emerging diseases: the case of severe acute respiratory syndrome (SARS). PMID- 12885152 TI - Altered affinity maturation in primary response to (4-hydroxy-3-nitrophenyl) acetyl (NP) after autologous reconstitution of irradiated C57BL/6 mice. AB - Immune responses developing in irradiated environment are profoundly altered. The memory anti-arsonate response of A/J mice is dominated by a major clonotype encoded by a single gene segment combination called CRIA. In irradiated and autoreconstituted A/J mice, the level of anti-ARS antibodies upon secondary immunization is normal but devoid of CRIA antibodies. The affinity maturation process and the somatic mutation frequency are reduced. Isotype switching and development of germinal centers (GC) are delayed. The primary antibody response of C57BL/6 mice to the hapten (4-hydroxy-3-nitrophenyl) acetyl (NP)-Keyhole Limpet Hemocyanin (KLH) is dominated by antibodies encoded by a family of closely related VH genes associated with the expression of the lambda1 light chain.We investigated the anti-NP primary response in irradiated and autoreconstituted C57BL/6 mice. We observed some splenic alterations as previously described in the irradiated A/J model. Germinal center reaction is delayed although the extrafollicular foci appearance is unchanged. Irradiated C57BL/6 mice are able to mount a primary anti-NP response dominated by lambda1 positive antibodies but fail to produce high affinity NP-binding IgG1 antibodies. Following a second antigenic challenge, irradiated mice develop enlarged GC and foci. Furthermore, higher affinity NP-binding IgG1 antibodies are detected. PMID- 12885153 TI - Phage display based cloning of proteins interacting with the cytoplasmic tail of membrane immunoglobulins. AB - The reduced quantity and quality of serum immunoglobulins (sIgs) in mutant mice expressing truncated cytoplasmic tails of IgE and IgG1 indicate an active role for the cytoplasmic domains of mIgG1 and mIgE. We used phage display technology to identify candidate proteins able to interact with the cytoplasmic tail of mIgE. Using a murine cDNA B cell library displayed on the surface of phage as prey and the 28 amino acid long cytoplasmic tail of IgE as bait, we isolated phage encoding the murine hematopoietic progenitor kinase 1 (HPK1). Surface plasmon resonance analysis measurements confirmed affinity of HPK1 to the mIgE cytoplasmic tail and revealed association to other immunoglobulin isotypes as well. Immunoprecipitation experiments, using lysates from two B cell lines expressing nitrophenyl (NP) specific mIgE molecules showed co-precipitation of IgE and HPK1. The interaction of HPK1 with the cytoplasmic domains of membrane immunoglobulins indicate an active role of the tails as part of an isotype specific signal transduction, independent from the Igalpha/Igbeta heterodimers, and may represent a missing link to upstream regulatory elements of HPK1 activation. PMID- 12885154 TI - Modulation of TNF-alpha secretion in peripheral blood mononuclear cells by cocoa flavanols and procyanidins. AB - Epidemiological reports have suggested that the consumption of foods rich in flavonoids is associated with a lower incidence of certain degenerative diseases, including cardiovascular disease. Flavanols and their related oligomers, the procyanidins CFP, isolated from cocoa can modulate the production and level of several signaling molecules associated with immune function and inflammation in vitro, including several cytokines and eicosanoids. To further elucidate the potential immuno-modulatory functions of flavanol-rich cocoa, the present investigation examined whether isolated CFP fractions (monomers through decamers) influence the secretion of tumor necrosis factor-alpha (TNF-alpha) from resting and phytohemagluttinin (PHA)-stimulated human peripheral blood mononuclear cells (PBMC). We used an in vitro culture system where PBMC from 14 healthy subjects were introduced to individual CFP fractions for 72 h prior to measuring the levels of TNF-alpha released. The intermediate-sized CFP fractions (tetramers through octamers) were the most active on resting cells, causing a 3-4 fold increase in TNF-alpha relative to media baseline. The monomers and dimers were the least stimulatory of the fractions tested, displaying a 42 and 31% increase, respectively, over media control, whereas the trimers, nonamers and decamers showed an intermediate stimulation of this cytokine. In the presence of PHA, the intermediate-sized CFP fractions again were the most active, enhancing TNF-alpha secretion in the range of 48-128% relative to the PHA control. The monomers and dimers were slightly inhibitory (-1.5 and -15%, respectively), while trimers, nonamers and decamers stimulated moderate increases in TNF-alpha levels (13, 19 and 15%, respectively). The above results lend support to the concept that CFP can be immunomodulatory. The stimulation of TNF-alpha secretion may contribute to the putative beneficial effects of dietary flavanoids against microbial infection and tumorigenesis. PMID- 12885155 TI - Kupffer cell-mediated recruitment of dendritic cells to the liver crucial for a host defense. AB - Tissue recruitment of dendritic cells (DCs) is essential for antigen presentation. When latex particulates were injected intravenously into rats, DC precursors were recruited to the liver. Propionibacterium acnes also induced the recruitment of definite mouse DC precursors. These DCs initially showed a selective binding to Kupffer cells. In the Kupffer cell-depleted rats, DCs could neither be recruited to the liver nor adhere to sinusoidal walls. Pretreatment with varied monosaccharides in vitro showed that sugar residues consisting of N acetylgalactosamine were necessary for this binding. Mouse DC precursors had CC chemokine receptor 1 and 5, while granulama tissues and rat Kupffer cells expressed the corresponding chemokine, macrophage inflammatory protein-lalpha. Recruited DC precursors phagocytosed latex or bacteria and some of them soon translocated to hepatic nodes and induced the immune response there. We conclude that after invasion of pathogens, Kupffer cells not only scavenge them but also recruit DCs/DC precursors via chemokine- and N-acetylgalactosamine-mediated interactions. The accelerated DC traffic and the presence of blood-lymph translocation would induce rapid and efficient immune responses and thus contribute to the local defense to antigens within liver tissues as well as systemic defense to blood-borne antigens. PMID- 12885156 TI - Allergic disease and autoimmune effectors pathways. AB - Allergy and autoimmunity result from dysregulation of the immune system. Until recently, it was generally accepted that the mechanisms that govern these disease processes are quite disparate; however, new discoveries suggest possible common pathogenetic effector pathways. This review illustrates the concomitant presentation of these conditions and the potential relationship or common mechanism in some cases, by looking at the key elements that regulate the immune response in both allergic and autoimmunite conditions: mast cells, antibodies, T cells, cytokines, and genetic determinants. The parallel appearance of allergic and autoimmune conditions in the some patients may reveal that such aberrations of the immune system have a common pathophysiologic mechanism. Mast cells, which play a key role in allergic reactions, and the wealth of inflammatory mediators they express, make it likely that they have profound effects on many autoimmune processes. Activation of protein kinases by inflammatory cytokines and environmental stresses may contribute to both allergic and autoimmune diseases. The presence of autoantibodies in some allergic conditions suggests an autoimmune basis for these conditions. Because of the central role T cells play in immune reactivity, the T-cell receptor (TCR) loci have long been considered important candidates for common disease susceptibility within the immune system such as asthma, atopy, and autoimmunity. Immunomodulation is the key to a successful treatment of allergic and autoimmune conditions. PMID- 12885157 TI - Involvement of GANP in B cell activation in T cell-dependent antigen response. AB - Adaptive immunity is dependent on proliferation of antigen-driven B cells for clonal expansion in germinal centers (GCs) against T cell-dependent antigens (TD Ag), accompanied with somatic hypermutation of variable-region gene and class switching of B cell antigen receptors. To study molecular mechanisms for B cell differentiation in GCs, we have identified and studied a 210kDa GANP protein expressed in GC-B cells. GANP has domains for MCM3-binding and RNA-primase activities and is selectively up-regulated in centrocytes surrounded with follicular dendritic cells (FDCs) upon immunization with TD-Ag in vivo and in B cells stimulated with anti-CD40 monoclonal antibody in vitro, which suggested that GANP plays a certain important role in the maturation of immunoglobulin or selection of B cells in GC during the immune response to TD-Ag. Since this up regulation has not been detected in T cells in GCs and in Concanavalin A stimulated T cells in vitro, selective function of GANP molecule on B cell proliferation and differentiation might exist. PMID- 12885158 TI - Functional tolerance is maintained despite proliferation of CD4 T cells after encounter with tissue-derived antigen. AB - Since negative selection in the thymus is incomplete, some self-reactive T cells are able to mature and seed the periphery. To study how these T cells interact following encounter with the self-protein they recognize in the periphery, we have developed an adoptive transfer system in which HEL-specific TCR transgenic CD4 T cells are transferred to mice expressing HEL protein in the pancreas under the control of the rat insulin promoter. Here we show that after adoptive transfer of HEL-specific T cells functional tolerance is maintained despite evidence that the T cells encounter and respond to pancreas-expressed antigen. Even the provision of an additional activation stimulus by peripheral immunization with HEL protein is insufficient to induce the T cells to cause autoimmune tissue injury. However, in the presence of blocking anti-CTLA-4-mAb, immunized adoptive transfer recipients rapidly developed diabetes. These data suggest that the CTLA-4 pathway regulates the pathogenicity of antigen-specific T cells following a peripheral activation stimulus. PMID- 12885159 TI - Exposure to environmental antigens induces the development of germinal centers in premature neonates. AB - The immune response of the neonate is poor and is dependent on passive immunity provided by maternal Ig. However, here we show that exposure of the neonate to environmental antigens induces a germinal center (GC) reaction. In the peripheral blood of premature infants one finds IgG class switched B cells expressing a selected V-gene repertoire. These data suggest that restrictions in the repertoire rather than immaturity of the immune system is responsible for the poor immune responses of the neonate. PMID- 12885161 TI - Transgene expression in the vegetative tissues of apple driven by the vascular specific rolC and CoYMV promoters. AB - The ability of the heterologous promoters, rolCP and CoYMVP, to drive expression of the gusA reporter gene in the vegetative tissues of apple (Malus pumila Mill.) has been studied using transgenic plants produced by Agrobacterium-mediated transformation. Replicate plants of each transgenic clone were propagated in soil to a uniform size and samples of leaf, petiole, stem, and root were taken for the measurement of beta-glucuronidase (GUS) activity by fluorometric assay. The levels of expression were compared with those in tissues of a representative clone containing the CaMV 35S promoter. These quantitative GUS data were related to the copy number of transgene loci assessed by Southern blotting. The CoYMV promoter was slightly more active than the rolC promoter, although both expressed gusA at a lower level than the CaMV 35S promoter. In clones containing the rolC promoter with multiple transgene loci, expression values were generally among the highest or lowest in the range. The precise location of GUS activity in each tissue was identified by staining of whole leaves and tissue sections with a chromogenic substrate. This analysis demonstrated that with both the rolC and CoYMV promoters the reporter gene activity was primarily localised to vascular tissues, particularly the phloem. Our results indicate that both promoters would be suitable to drive the expression of transgenes to combat pests and diseases of apple that are dependent on interaction with the phloem. PMID- 12885160 TI - Fish can be first--advances in fish transgenesis for commercial applications. AB - Over the past 15 years researchers have generated stable lines of several species of transgenic fish important for aquaculture. 'All-fish' growth hormone (GH) gene constructs and antifreeze protein (AFP) genes have been successfully introduced into the fish genome resulting in a significant acceleration of growth rate and an increase in cold and freeze tolerance. However, neither gene modification is completely understood; there are still questions to be resolved. Expression rates are still low, producing variable growth enhancement rates and less than desired levels of freeze resistance. Transgene strategies are also being developed to provide improved pathogen resistance and modified metabolism for better utilization of the diet. Additional challenges are to tailor the genetically modified fish strains to prevent release of the modified genes into the environment. PMID- 12885162 TI - An efficient procedure to stably introduce genes into an economically important pulp tree (Eucalyptus grandis x Eucalyptus urophylla). AB - Regeneration problems are one of the main limitations preventing the wider application of genetic engineering strategies to the genus Eucalyptus. Seedlings from Eucalyptus grandis x Eucalyptus urophylla were selected according to their regeneration (adventitious organogenesis) and transformation capacity. After in vitro cloning, the best genotype of 250 tested was transformed via Agrobacterium tumefaciens. A cinnamyl alcohol dehydrogenase (CAD) antisense cDNA from Eucalyptus gunnii was transferred, under the control of the 35S CaMV promoter with a double enhancer sequence, into a selected genotype. According to kanamycin resistance and PCR verification, 120 transformants were generated. 58% were significantly inhibited for CAD activity, and nine exhibited the highest down regulation, ranging from 69 to 78% (22% residual activity). Southern blot hybridisation showed a low transgene copy number, ranging from 1 to 4, depending on the transgenic line. Northern analyses on the 5-16 and 3-23 lines (respectively one and two insertion sites) demonstrated the antisense origin of CAD gene inhibition. With respectively 26 and 22% of residual CAD activity, these two lines were considered as the most interesting and transferred to the greenhouse for further analyses. PMID- 12885163 TI - High efficiency production of germ-line transgenic Japanese medaka (Oryzias latipes) by electroporation with direct current-shifted radio frequency pulses. AB - Although there have been several studies showing the production of transgenic fish through electroporation techniques, success rates have been low and few studies show germ-line integration and expression. When electroporation has been successful, the device used is no longer commercially available. The goal of this experiment was to find an alternative efficient method of generating transgenic Japanese medaka (Oryzias latipes) using a commercially available electroporation device. The Gene Pulser II and RF module (Bio-Rad Laboratories, USA), along with two reporter gene constructs, were used. In contrast to other electroporation devices, which are based on a single pulse with exponential decay or square wave technology, the Gene Pulser II incorporates a direct current (DC)-shifted radio frequency (RF) signal. With this technique, over 1000 embryos can be electroporated in less than 30 min. The plasmid pCMV-SPORT-beta-gal (Invitrogen, USA) was used in the supercoiled form to optimize parameters for gene transfer into single-celled embryos, and resulted in up to 100% somatic gene transfer. Similar conditions were used to generate fish transgenic for both the pCMV-EGFP plasmid (Clontech, USA) and a cytomegalovirus (CMV) driven phytase-EGFP construct. The conditions used were a voltage of 25 V, a percent modulation of 100%, a radio frequency of 35 kHz, a burst duration of 10 ms, 3 bursts, and a burst interval of 1.0 s. Seventy percent of the embryos electroporated with the pCMV-EGFP construct survived to sexual maturity, and of those, 85% were capable of passing the transgene on to their offspring. Transgenic second generation back crossed (BC2) fry were subjected to Southern blot analysis, which confirmed germ line integration, and observation for green fluorescence protein, which confirmed protein expression. DC-shifted RF pulses are effective and efficient in the production of transgenic medaka, and germ-line integration and expression can be achieved without linearization of the transgene vector. PMID- 12885164 TI - Homology-dependent DNA transfer from plants to a soil bacterium under laboratory conditions: implications in evolution and horizontal gene transfer. AB - DNA transfer was demonstrated from six species of donor plants to the soil bacterium, Acinetobacter spp. BD413, using neomycin phosphotransferase (nptII) as a marker for homologous recombination. These laboratory results are compatible with, but do not prove, DNA transfer in nature. In tobacco carrying a plastid insertion of nptII, transfer was detected with 0.1 g of disrupted leaves and in oilseed rape carrying a nuclear insertion with a similar quantity of roots. Transfer from disrupted leaves occurred in sterile soil and water, without the addition of nutrients. It was detected using intact tobacco leaves and intact tobacco and Arabidopsis plants in vitro. Transfer was dose-dependent and sensitive to DNase, and mutations in the plant nptII were recovered in receptor bacteria. DNA transfer using intact roots and plants in vitro was easily demonstrated, but with greater variability. Transfer varied with plant genome size and the number of repeats of the marker DNA in the donor plant. Transfer was not detected in the absence of a homologous nptII in the receptor bacteria. We discuss these results with reference to non-coding DNA in plant genomes (e.g., introns, transposons and junk DNA) and the possibility that DNA transfer could occur in nature. PMID- 12885165 TI - A sequential approach to risk assessment of transgenic plants expressing protease inhibitors: effects on nontarget herbivorous insects. AB - Protease inhibitors expressed in transgenic plants can provide enhanced levels of resistance to important pest species. A sequential approach for testing the effects of protease inhibitor-expressing crops on nontarget herbivorous insects has been developed. The approach consists of five tiers. The first two tiers comprise the selection phase. In tier one, field surveys are used to characterise the nontarget invertebrate fauna of a crop. In tier 2, histochemical assays are used to identify the subset of herbivores with a particular class of digestive proteolytic enzymes. In the assessment phase a combination of laboratory 'worst case scenario' studies (tier 3) and controlled environment or small-scale field trials (tier 4) are used to evaluate the impact of the protease inhibitor expressing plants on the selected nontarget species. In the final tier, field trials are used to compare the relative effect of transgenic plants and current management practices, such as pesticide use, on selected species. The first four tiers of the approach are described using potatoes expressing cystatins, a family of cysteine proteinase inhibitors, as an example. Although the plants have enhanced levels of resistance to potato cyst nematodes (PCN), Globodera pallida and Globodera rostochiensis, the results establish that they have negligible impact on the nontarget herbivorous insect, Eupteryx aurata. PMID- 12885166 TI - Isolation of murine embryonic stem and embryonic germ cells by selective ablation. AB - The isolation of murine embryonic stem (ES) cells has been almost exclusively from the 129 mouse strain. Other mouse strains, such as CBA, have proven refractory to ES cell isolation by conventional means. We previously reported the isolation of 87.5% CBA ES lines by selective ablation of differentiating cells (McWhir et al., 1996). Here, we report the isolation of ES and EG cells from 94% CBA embryos hemizygous and homozygous for a neomycin-resistance transgene under the transcriptional control of the Oct3/4 promoter (Oct/neo). Since expression of the Oct/neo transgene only confers drug resistance to undifferentiated cells of the inner cell mass, selection results in the ablation of differentiating cells from the culture. The efficiency of ES isolation by selective ablation in homozygotes is twice that in heterozygotes. ES isolation frequency in permissive strain 129 embryos is enhanced by treatment with an inhibitor of the extracellular-signal-regulated kinase (ERK) pathway but this effect is not sufficient to permit ES isolation from the CBA strain. PMID- 12885167 TI - Transgene organisation in potato after particle bombardment-mediated (co )transformation using plasmids and gene cassettes. AB - Protocols for efficient co-transformation of potato internodes with genes contained in separate plasmids or gene cassettes (i.e., linear PCR fragments comprising a promoter-gene-terminator) using particle bombardment were established. Twenty-eight out of 62 (45%) and 11 out of 65 (17%) plants transformed with a plasmid containing the selectable marker contained one and two additional non-selected genes, respectively. When gene cassettes were used in transformation, six out of eight plants were co-transformed. Expression analysis showed that 75-80% of the plants transformed with two transgenes expressed both of them, irrespective of the use of plasmids or gene cassettes. Thirty-eight plants containing the gusA reporter-gene and the nptII selectable-marker have been characterised with respect to the molecular organisation of the donor DNAs. Seventeen out of 49 (35%) gusA sites of integration contained one copy of the gene. Only 11 gusA sites (22%) were linked to the site of integration of the selectable marker. When one site of integration contained several copies of the transgene, a predominance of 3'-3' inverted re-arrangement repeats was observed. PMID- 12885168 TI - Co-expression of a modified maize ribosome-inactivating protein and a rice basic chitinase gene in transgenic rice plants confers enhanced resistance to sheath blight. AB - Chitinases, beta-1,3-glucanases, and ribosome-inactivating proteins are reported to have antifungal activity in plants. With the aim of producing fungus-resistant transgenic plants, we co-expressed a modified maize ribosome-inactivating protein gene, MOD1, and a rice basic chitinase gene, RCH10, in transgenic rice plants. A construct containing MOD1 and RCH10 under the control of the rice rbcS and Act1 promoters, respectively, was co-transformed with a plasmid containing the herbicide-resistance gene bar as a selection marker into rice by particle bombardment. Several transformants analyzed by genomic Southern-blot hybridization demonstrated integration of multiple copies of the foreign gene into rice chromosomes. Immunoblot experiments showed that MOD1 formed approximately 0.5% of the total soluble protein in transgenic leaves. RCH10 expression was examined using the native polyacrylamide-overlay gel method, and high RCH10 activity was observed in leaf tissues where endogenous RCH10 is not expressed. R1 plants were analyzed in a similar way, and the Southern-blot patterns and levels of transgene expression remained the same as in the parental line. Analysis of the response of R2 plants to three fungal pathogens of rice, Rhizoctonia solani, Bipolaris oryzae, and Magnaporthe grisea, indicated statistically significant symptom reduction only in the case of R. solani (sheath blight). The increased resistance co-segregated with herbicide tolerance, reflecting a correlation between the resistance phenotype and transgene expression. PMID- 12885169 TI - Increased efficiency of transgenic livestock production. AB - Production of transgenic livestock by pronuclear microinjection of DNA into fertilized zygotes suffers from the compounded inefficiencies of low embryo survival and low integration frequencies of the injected DNA into the genome. These inefficiencies are one of the major obstacles to the large-scale use of pronuclear microinjection techniques in livestock. We investigated exploiting the properties of recombinase proteins that allow them to bind DNA to generate transgenic animals via pronuclear microinjection. In theory, the use of recombinase proteins has the potential to generate transgenic animals with targeted changes, but in practice we found that the use of RecA recombinase coated DNA increases the efficiency of transgenic livestock production. The use of RecA protein resulted in a significant increase in both embryo survival rates and transgene integration frequencies. Embryo survival rates were doubled in goats, and transgene integration was 11-fold higher in goats and three-fold higher in pigs when RecA protein-coated DNA was used compared with conventional DNA constructs without RecA protein coating. However, a large number of the transgenic founders generated with RecA protein-coated DNA were mosaic. The RecA protein coating of DNA is straightforward and can be applied to any species and any existing microinjection apparatus. These findings represent significant improvements on standard pronuclear microinjection methods by enabling the more efficient production of transgenic livestock. PMID- 12885170 TI - The Saccharomyces cerevisiae chitinase, encoded by the CTS1-2 gene, confers antifungal activity against Botrytis cinerea to transgenic tobacco. AB - The Saccharomyces cerevisiae chitinase, encoded by the CTS1-2 gene has recently been confirmed by in vitro tests to possess antifungal abilities. In this study, the CTS1-2 gene has been evaluated for its in planta antifungal activity by constitutive overexpression in tobacco plants to assess its potential to increase the plant's defence against fungal pathogens. Transgenic tobacco plants, generated by Agrobacterium-mediated transformation, showed stable integration and inheritance of the transgene. Northern blot analyses conducted on the transgenic tobacco plants confirmed transgene expression. Leaf extracts from the transgenic lines inhibited Botrytis cinerea spore germination and hyphal growth by up to 70% in a quantitative in vitro assay, leading to severe physical damage on the hyphae. Several of the F1 progeny lines were challenged with the fungal pathogen, B. cinerea, in a detached leaf infection assay, showing a decrease in susceptibility ranging from 50 to 70%. The plant lines that showed increased disease tolerance were also shown to have higher chitinase activities. PMID- 12885171 TI - The endosperm and the embryo of Arabidopsis thaliana are independently transformed through infiltration by Agrobacterium tumefaciens. AB - Several experiments had indicated that in planta transformation of Arabidopsis thaliana by Agrobacterium involves the female germ line. In order to identify the precise stage at which transformation occurs we have monitored expression of a gusA reporter gene in the two products of the double fertilization of infiltrated plants. The plantlets and the remaining endosperm of seeds were separately tested after germination. It appeared that in the majority of cases only the plantlet or the endosperm were transformed. Based on transformation with two vectors borne by two different Agrobacterium strains, the minority of 'co-transformed' plantlets and endosperm can be explained by simultaneous but independent transformation events. These results indicate that mature female gametes could be the targets of T-DNA. PMID- 12885172 TI - A simple genotyping assay for the Hprt null allele in mice produced from the HM-1 and E14TG2a mouse embryonic stem cell lines. PMID- 12885173 TI - Universal PCR genotyping assay that achieves single copy sensitivity with any primer pair. PMID- 12885174 TI - A rapid and simple PCR-based method for analysis of transgenic fish using a restricted amount of fin tissue. AB - The protocol described in this paper offers a simple and rapid method for PCR analysis of transgenes using a restricted amount of fin tissue from small-sized transgenic fish. A simple preparation of fin lysate using a buffer containing a low concentration of an ionic detergent, SDS (0.01%), followed by neutralization with a second buffer containing higher concentrations of non-ionic detergents NP40 (2%) and Tween 20 (2%) consistently provides a reliable quantity of high quality DNA template for PCR amplification of transgenes. Based on this protocol, transgenic fish can be clearly distinguished from non-transgenic fish using PCR in a rapid and reproducible manner. Tedious DNA purifications are avoided while fidelity of amplification and efficient identification of transgenic fish are maintained. PMID- 12885175 TI - Correlation beween AAI-index and the BIS-index during propofol hypnosis: a clinical study. AB - OBJECTIVE: To determine the degree of linearity and correlation between the anaesthetic depth indices BIS and AAI over a wide range of hypnotic depth using propofol. METHODS: 20 ASA I patients were studied during propofol induction. Co induction with 0.05 mg fentanyl and 30 mg propofol iv before initiation of the study. Thereafter repeated doses of propofol 0.5 mg/kg iv. every minute until BIS < 30. Loss of responsiveness to verbal command was determined by repeated loud commands to the patient. BIS (Aspect 2000 XP, BIS algorithm 4.0, system rev. 3.12, Aspect Medical Systems; Natick, MA, U.S.A.) and AAI-index (A-Line Auditory Evoked Potential Monitor, version 1.4, Danmeter A/S; Odense, Denmark) were determined simultaneously (n = 15). BIS alone without acoustical stimulation was studied in a control group (n = 5). MAIN RESULTS: Both indices decreased with increasing dose, and there was a high correlation between the two (r2 = 0.82). The indices showed however different values and while BIS were quite linear, the AAI-index had a more on-offb ehaviour. CONCLUSION: The AAI-index correlates with the BIS-index during propofol hypnosis in the absence of surgical stimulation. Neither the BIS-index, heart rate, nor systemic blood pressure were influenced by the acoustical stimulation from the A-line monitor. Both indices decreased in relation to increasing doses of propofol, but the AAI-index was lower both before becoming unconscious, during transition to unconsciousness, and during the deeper levels of sedation. The AAI-index lacks linearity at both very low and higher levels of propofol sedation with a nearly on-off behaviour for wakefulness vs hypnosis. PMID- 12885176 TI - A better landmark for positioning a central venous catheter. AB - BACKGROUND: Improper positioning of central venous catheters (CVCs) can lead to erosion through the superior vena cava (SVC) or right atrium (RA) and pericardial tamponade. It is widely recommended that the tip of CVCs be placed above the heart or the pericardial reflection. The purpose of this study was to identify an easily recognized landmark to allow identification of the proximal extent of the pericardial reflection on a routine chest radiograph (CXR). METHODS: We analyzed the computerized tomograms of the chest from 97 adults to evaluate the relationship between the pericardial reflection, SVC, carina, and right mainstem bronchus. Correlations between demographic data and length of SVC or pericardial reflection were sought. RESULTS: The mean length of the SVC was 6.5 cm. The pericardial reflection covered an average of 3.6 cm of the distal SVC. The carina was a mean of 1.3 cm below the mid-point of the SVC and 0.7 cm below the pericardial reflection. There was no significant correlation between SVC or pericardial length and either age, height, or weight. CONCLUSIONS: The distal half of the SVC lies within the pericardial reflection, and the upper limit of the pericardial reflection is slightly above the level of the carina. These landmarks are useful for determining proper position of the tip of a CVC on CXR. PMID- 12885177 TI - Computerize anesthesia record keeping in thoracic surgery--suitability of electronic anesthesia records in evaluating predictors for hypoxemia during one lung ventilation. AB - OBJECTIVE: The aim of this retrospective study was to assess the suitability of routine data gathered with a computerized anesthesia record keeping system in investigating predictors for intraoperative hypoxemia (SpO2 < 90%) during one lung ventilation (OLV) in pulmonary surgery. METHODS: Over a four-year period data of 705 patients undergoing thoracic surgery (pneumonectomy: 78; lobectomy: 292; minor pulmonary resections: 335) were recorded online using an automated anesthesia record-keeping system. Twenty-six patient-related, surgery-related and anesthesia-related variables were studied for a possible association with the occurrence of intraoperative hypoxemia during OLV. Data were analyzed using univariate and multivariate (logistic regression) analysis (p < 0.05). The model's discriminative power on hypoxemia was checked with a receiver operating characteristic (ROC) curve. Calibration was tested using the Hosmer-Lemeshow goodness-of-fit test. RESULTS: An intraoperative incidence of hypoxemia during OLV was found in 67 patients (9.5%). Using logistic regression with a forward stepwise algorithm, body-mass-index (BMI, p = 0.018) and preoperative existing pneumonia (p = 0.043) could be detected as independent predictors having an influence on the incidence of hypoxemia during OLV. An acceptable goodness-of-fit could be observed using cross validation for the model (C = 8.21, p = 0.370, degrees of freedom, df 8; H = 3.21, p = 0.350, df 3), the discriminative power was poor with an area under the ROC curve of 0.58 [0.51-0.66]. CONCLUSIONS: In contrast to conventional performed retrospective studies, data were directly available for analyses without any manual intervention. Due to incomplete information and imprecise definitions of parameters, data of computerized anesthesia records collected in routine are helpful but not satisfactory in evaluating risk factors for hypoxemia during OLV. PMID- 12885178 TI - A preliminary evaluation of a new derived EEG index monitor in anesthetized patients. AB - BACKGROUND: An electroencephalogram (EEG) monitoring device, recently developed by Nicolet Biomedical, analyzes both high and low EEG frequencies. A processed derivative is obtained and displayed graphically and numerically on a monitor and may be used during anesthesia to indicate anesthetic "depth." However, radio frequency interference from electrical equipment typically used in the operating room has the potential to interfere with the analysis of the high frequency components of the EEG. OBJECTIVE: The objective of this study was to determine the ability of the Nicolet EEG monitoring device to function satisfactorily and effectively in the operating arena when used for anesthetized patients. METHODS: A total of 40 patients undergoing surgery with general anesthesia were monitored using a prototype, processed EEG monitoring device. The device was used beginning prior to induction and continuing until emergence from anesthesia. RESULTS: Analysis was performed on 38 of the 40 patients. Brief interruption of the derived EEG Index occurred during use of the monopolar electrosurgical unit (ESU) as well as during stimulation of the facial nerve (used to monitor pharmacologic muscle relaxation). Derived EEG Index interruption was also noted during some movements of the patient's head or the EEG electrode wires. Changes in the derived EEG Index value accompanied changes in inhaled concentration of anesthetics and, in one patient, change in the derived EEG Index preceded patient movement that was not heralded by the typical cardiovascular indications of "light" anesthesia. CONCLUSIONS: The derived EEG Index monitoring system evaluated in this study functioned satisfactorily in the operating room setting in patients undergoing general anesthesia. Interference from ESU, facial nerve stimulation, and movement of the electrode wires caused brief interruptions of the derived EEG Index display and did not affect its utility in monitoring brain activity during anesthesia. PMID- 12885179 TI - Temporal patient state characterization using Iterative Order and Noise (ION) estimation: applications to anesthesia patient monitoring. AB - OBJECTIVE: As more sensors are added to increasingly technology-dependent operating rooms (OR), physicians such as anesthesiologists must sift through an ever-increasing number of patient parameters every few seconds as part of their OR duties. To the extent these many parameters are correlated and redundant, manually monitoring all of them may not be an optimal physician strategy for assessing patient state or predicting future changes to guide their actions. METHODS: The method is illustrated by application to seventy-six anesthetized patients for which thirty-two fundamental and derived variables were recorded at 20-second intervals. The Iterative Order and Noise estimation algorithm (ION) estimated the noise on each parameter. The performance of principal components analysis (PCA) was improved by normalizing the noise estimated by ION to unity. A linear regression of the resulting seven high signal-to-noise ratio principal components (PC's) predicted tachycardia 140 seconds in advance. RESULTS: ION estimated the noise on each parameter with sufficient accuracy to increase the number of significant PC's from two to seven, all of which had identifiable physiological correlates. The resulting receiver operating characteristic (ROC) suggested that a 70 percent prediction rate with 5 percent false alarms could be achieved. CONCLUSIONS: This paper illustrates the use of ION to improve significantly the performance of PCA in the efficient representation of patient state and in improving the performance of linear predictors of clinically significant parameters. PMID- 12885180 TI - Unconsciousness indication using time-domain parameters extracted from mid latency auditory evoked potentials. AB - OBJECTIVE: The mid-latency auditory evoked potential (MLAEP) has been used to indicate depth of anaesthesia, and is usually analysed in time-domain. This work compares three techniques: Wave Deformation Parameters (PDO), Auditory Evoked Potential Index (AEPidx) and an automatic Nb-wave latency estimator (Nb), in the assessment of unconsciousness onset based on EEG under auditory stimulation. METHODS: Ten normal adult volunteers, under no pre-anaesthetic drug administration, received propofol during two successive periods of 45 min each one (3 mg/Kg/h and 9 mg/Kg/h), being the EEG collected from 10 min previous to infusion beginning until the subjects woke up. From the time-evolution of MLAEP (averaging of successive sets of 1000 epochs) all the parameters were compared to thresholds (unconsciousness onset indication time) and the results were compared to the instant of pressing interruption of a soft-touch switch, when one assumed the volunteer became unconscious. RESULTS: The Wilcoxon Signed-Rank test points equivalence between each of the parameters and the switch for, say, alpha = 5%. Bland-Altman diagrams revealed that the Attenuation-PDO has better agreement to the switch than Nb and AEPidx. CONCLUSION: The results suggest that, at least to indicate unconsciousness, the most reliable effect of the anaesthetic drug on MLAEP would be the amplitude attenuation. Despite the high dependence on noise due to its time-domain basis, the Attenuation-PDO seems to be adequate to assess depth of anaesthesia. PMID- 12885181 TI - Closed-loop control if the inspired fraction of oxygen in mechanical ventilation. AB - OBJECTIVE: Supplemental oxygen treatment of patients on mechanical ventilation is crucial in maintaining the patients' oxygen levels in the normal range. The purpose of this study was to evaluate the effectiveness of a closed-loop controller for automatic adjustment of the fraction of inspired oxygen, FIO2. More specifically, the aim of the study was to assess the robustness of the controller in correcting hypoxemia as well as its effectiveness in prevention of hyperoxemia and oxygen toxicity. METHODS: The microprocessor-based feedback control system combines a rapid control algorithm with a proportional-integral derivative (PID) control procedure to automatically adjust FIO2. The system is designed to prevent hypoxemia by applying a stepwise control procedure in response to rapid declines in arterial oxygen saturation while fine-tuning FIO2 and avoiding hyperoxemia by resuming to the PID control procedure when appropriate. The system includes a sophisticated safeguard unit which is designed to communicate any oxygenation problems or measurement artifacts to the medical personnel while keeping FIO2 at a safe and sufficiently high level. The control system has been tested by using computer simulations as well as animal studies. RESULTS: In response to different disturbances, the arterial oxygen saturation returned to the normal safe range within less than 20 seconds, thereby avoiding any untoward effects of hypoxemia. Under steady state conditions, the variations in arterial oxygen saturation were kept within +/- 3% of the mean value. The controller corrected hypoxemia within seconds while preventing hyperoxemia, rejecting artifacts, and minimizing exposure to high concentrations of oxygen. CONCLUSION: The results of the study attest to the reliability of the proposed closed-loop control scheme for automatic adjustment of FIO2. Further evaluation of the controller will require testing the effectiveness of the system on different patient groups. PMID- 12885182 TI - Biasing effect of the electromyogram on BIS: a controlled study during high-dose fentanyl induction. AB - OBJECTIVE: A biasing effect of the electromyogram (EMG) on the Bispectral Index (BIS) may explain discrepancies in previous studies assessing BIS in the presence of neuromuscular activity. Our aims were: to evaluate variations of BIS in the presence of high EMG activity associated with muscular rigidity after administration of high-dose fentanyl; to compare muscular rigidity, as measured by the EMG variable of the BIS monitor, in patients who were administered two different dosages of fentanyl at induction of cardiac anaesthesia. METHODS: 26 patients undergoing CABG surgery, after premedication with morphine 0.15 mg/kg, were randomized to receive either fentanyl 50 mcg/kg (group F) or fentanyl 10 mcg/kg plus etomidate 0.2 mg/kg (group EF). The induction dose was administered over 2 minutes. Patients were manually ventilated with O2 via face mask. Five minutes after induction was complete, patients were clinically assessed using the Responsiveness portion of the Observer's Assessment of Alertness/Sedation scale (OAAS). Haemodynamic data were recorded and arterial blood samples obtained at the time of OAAS observation. Patients were administered a neuromuscular blocking agent only after the OAAS assessment. BIS (3.4) was recorded from an A-2000 EEG monitor (Aspect Medical Systems) using disposable sensors (BIS Sensor, Aspect Medical Systems) applied per manufacturer's instructions. Data were recorded on a PC for off-line analysis. RESULTS: At the time of OAAS observation, mean (95% CI) BIS in group F was 85 (77-92) compared to 67 (56-79) in group EF (p = 0.01). Similarly, mean (95% CI) EMG was 50 dB (45-56) in F and 41 dB (35-47) in EF (p = 0.01). Correlation between BIS and EMG was very high (r2 = 0.88). OAAS scores were significantly higher in group F (p = 0.03). Non significant correlation was observed between BIS and OAAS scores (r2 = 0.32, p = 0.1). Backward stepwise multiple regression analysis including EMG, pH, CO2, O2 and OASS scores showed EMG as strong predictor of BIS (p < 0.0001, r2 = 0.7). Regression of EMG against BIS yielded the equation: BIS = 3.7 + (1.6 x EMG). CONCLUSION: During fentanyl induced muscular rigidity BIS recordings reflect EMG variations. When assessing BIS in the absence of neuromuscular blockade, it is necessary to evaluate the effect of EMG on BIS before making conclusions about depth of sedation. Fentanyl induced rigidity appears to be a dose-related phenomenon which the EMG variable of BIS 3.4 is able to quantify. PMID- 12885183 TI - Sex ratio of Plasmodium falciparum gametocytes in inhabitants of Dielmo, Senegal. AB - An epidemiological survey was conducted during a 4-month period of intense malaria transmission in Dielmo, a holoendemic Senegalese village. Two thick blood smears per inhabitant were collected weekly. The sex ratio of Plasmodium falciparum gametocytes (gamete precursors) was studied in 50 gametocyte carriers. All age classes were represented (mean 19.7 years; range: 2 months-75 years); 42 (84%) of them did not receive antimalarial treatment. Overall 668 thick smears were examined until 100 gametocytes had been counted or for 40 min. A total of 11204 gametocytes were observed with a mean sex ratio of 0.346 (95% CI 0.317 0.374), i.e. 2.89 females per 1 male. Among the 284 thick smears in which at least 10 gametocytes were observed, the mean percentage of male gametocytes was 27.8%, with a range of 0-82%. Great variability was observed between gametocyte carriers and also between thick smears from the same gametocyte carrier. A multivariate analysis was performed which highlighted the fact that only 2 variables had a significant effect on the sex ratio. Anaemia was associated with an increased percentage of males (Prevalence Rate Ratio [PPR] of male gametocytes was multiplied by 1.65 if haematocrit rate < 32%) and a wave of gametocytes was associated with an increased percentage of female gametocytes (PRR was multiplied by 0.48 during the peak of gametocytaemia and for the 2 weeks following this peak). The variables without significant effect on sex ratio were: age, sex, clinical status and sickle cell trait status of the gametocyte carrier, density of asexual parasites, quinine treatment, and gametocyte density (when taking account of its waves). These results are discussed in regard of possible differential production, mortality or sequestration of one gametocyte sex and selective advantages for the transmission of parasites. PMID- 12885184 TI - The influence of malaria parasite genetic diversity and anaemia on mosquito feeding and fecundity. AB - Studies of invertebrate-parasite interactions frequently report that infection reduces host fecundity. The extent of the reduction is likely to be determined by a wide range of host and parasite factors. We conducted a laboratory experiment to evaluate the role of parasite genetics and infection genetic diversity on the fecundity of mosquitoes carrying malaria parasites. The malaria vector Anopheles stephensi was infected with either of 2 different genotypes of the rodent malaria parasite Plasmodium chabaudi, or by a mixture of both. Mixed genotype infections reduced mosquito fecundity by 20%, significantly more than either of the 2 single genotype infections. Mixed genotype infections were associated with high gametocyte densities and anaemia in mice, both of which were correlated with reduced bloodmeal size in mosquitoes. Bloodmeal size was the most important predictor of mosquito fecundity; the presence and number of parasites had no direct effect. Parasite density influenced the propensity of mosquitoes to feed on infected mice, with a higher percentage of mosquitoes taking a meal as asexual parasite and gametocyte density increased. Thus mosquitoes may preferentially feed on hosts who will most impair their fecundity. PMID- 12885185 TI - The morphological discrimination of microfilariae of Onchocerca volvulus from Mansonella ozzardi. AB - There is no published account which allows the morphological discrimination of microfilariae of Onchocerca volvulus and M. ozzardi from each other. However, they occur together in parts of Brazil and Venezuela, and presumably there is always the possibility that migration could establish new sympatric populations in the future. The objective of this study was to evaluate simple morphological characters that might be used for species-diagnosis of microfilariae. The conclusions were that the location of microfilariae in the blood or skin, the body size and the nucleation of the nerve ring are expected to be useful first indications of species identity, but cannot be used for confident diagnosis. The structure of the cephalic armature (stained with alcian blue) seems to be species specific, but is of limited application because it is often difficult to see. However, the pattern of nucleation of the tail (as expressed by the ratio of the length of the terminal nucleus compared with the length of the tail space) is distinctive and is expected to be diagnostic. PMID- 12885186 TI - Age-dependent survival and infectivity of Schistosoma mansoni cercariae. AB - The age dependency of the mortality, spontaneous de-tailing and infectivity of cercariae of Schistosoma mansoni has been determined at 25 degrees C. Infectivity was assessed with respect to stratum corneum-like differentiated human keratinocyte cultures (validated by comparison with fresh human skin samples) and displayed a complex age-dependent pattern. From 1 to 9 h post-emergence cercariae showed a plateau of maximal infectivity (around 90% attachment). Thereafter, infectivity declined. Immediately after release, infectivity at around 60% was significantly lower than the plateau values and this could be an adaptation for spatial dispersal of cercariae. Age-dependent patterns of cercarial mortality and spontaneous de-tailing closely mirrored the infectivity pattern except in relation to the low initial infectivity value. These findings suggest that, at a population level, the age-dependent decline in cercarial infectivity towards human skin is essentially driven by cercarial mortality. The recently described phenomenon of delayed tail loss (DTL) in S. mansoni cercariae infecting human skin is confirmed in the present study. For cercariae aged up to 13.5 h post emergence, 90% or more of invading cercariae took their tails with them into the keratinocyte culture. The infection dynamics described in this study suggest that diurnally shed S. mansoni cercariae, with peak emergence around mid-day, will have near maximal infectivity towards humans in contact with water through all remaining daylight hours in the tropics. PMID- 12885187 TI - Structure and organization of the mitochondrial genome of the canine heartworm, Dirofilaria immitis. AB - This study determined the complete mitochondrial (mt) genome sequence of the canine heartworm, Dirofilaria immitis, and compared its structure, organization and other characteristics with Onchocerca volvulus and other secernentean nematodes. The D. immitis mt genome is 13814 bp in size and contains 36 of the 37 genes typical of metazoan organisms, and lacks the ATP synthetase subunit 8 gene. All of the genes are transcribed in the same direction. For the entire genome, the nucleotide contents are approximately 55% (T), approximately 19% (each for A and G) and approximately 7% (C), which is very similar to those of the protein coding genes. In the latter genes, most (approximately 69%) third codon positions have a T, but rarely (approximately 1-9%) have an A or a C. The C content (8-12%) is higher at the first and second codon positions compared with the third position (approximately 1%). These nucleotide biases have a significant effect on the codon usage patterns and, thus, on the amino acid composition of the proteins. The mt genome organization of D. immitis is essentially the same as that of O. volvulus, but is distinctly different from other secernentean nematodes sequenced thus far. Irrespective of transpositions of transfer RNA (trn) genes and the non-coding, AT-rich region, there are 4 gene- or gene block translocations between the mt genome of D. immitis and those of Caenorhabditis elegans, Ascaris suum and the 2 human hookworms, Ancylostoma duodenale and Necator americanus. For D. immitis, the 22 trn genes have secondary structures typical of other secernentean nematodes, and possess a TV-replacement loop instead of a TpsiC arm and loop. Like O. volvulus, the mt trnK and trnP of D. immitis use the anticodons CUU and AGG, whereas in other nematodes, UUU and UGG are employed, respectively. Also, the secondary structures of the 2 ribosomal RNA (rrn) genes are similar to the models for other nematodes. Overall, the availability of the complete D. immitis mt genome sequence provides a resource for future studies of the comparative mt genomics and of the population genetics and/or phylogeny of parasitic nematodes. PMID- 12885188 TI - Analysis of the beta-tubulin codon 200 genotype distribution in a benzimidazole susceptible and -resistant cyathostome population. AB - To study the prevalence of the polymorphism in position 200 of the beta-tubulin gene in the mechanism of benzimidazole (BZ) resistance in cyathostomes of horses, an allele-specific PCR was used to detect the genotype of individuals of BZ susceptible and BZ-resistant populations. The molecular analysis of 100 adults recovered from an anthelmintic-naive horse revealed 80% homozygous TTC/TTC individuals, 17% heterozygous TTC/TAC and 3% homozygous TAC/TAC. A naturally infected horse was treated with increasing fenbendazole (FBZ) dosages to select a BZ-resistant population of cyathostomes. The PCR based analysis of 3rd-stage larvae (L3) during the experiment revealed a decrease of the homozygous TTC/TTC genotype and an increase in heterozygous TTC/TAC and homozygous TAC/TAC individuals. After treatment 42.3% of the adults (n=104) were homozygous TTC/TTC, 55.8% were heterozygous TTC/TAC and only 1.9% showed the homozygous genotype TAC/TAC. The results of the molecular analysis lead to the proposal that polymorphism within codon 200 is not the only reason for the development of BZ resistance in small strongyles. PMID- 12885189 TI - The effect of inulin and sugar beet fibre on oesophagostomum dentatum infection in pigs. AB - The present study was designed to assess the role of inulin and sugar beet fibres (SBF) on adult O. dentatum in growing pigs. Four experimental diets were formulated based on barley flour with added insoluble fibre from oat husk (Diet 1), a pure carbohydrate source inulin (Diet 2), soluble fibre from sugar beet fibre (SBF) with a high proportion of soluble fibre components (Diet 3) or inulin plus SBF (Diet 4). Thirty-two 10-week-old pigs were divided randomly into 4 groups each of 8 pigs. After 3 weeks adaptation on Diet 1 all pigs were infected with a single dose of 6000 L8 O. dentatum. At week 7 p.i. one group was switched to Diet 2, another group to Diet 3 and another group to Diet 4. The remaining 8 pigs continued on Diet 1 until the end of the experiment and served as controls. At week 13, all pigs were necropsied and their worm burdens determined. The worm recoveries from the pigs on the inulin supplemented diet (Diet 2) were reduced by 97% compared to the controls (Diet 1). Further, the inulin-fed pigs exhibited markedly reduced faecal egg counts. The pigs on inulin plus SBF diet (Diet 4) and on SBF diet (Diet 3) had 86% and 70% adult worm reductions compared with the controls, respectively. The results from this study indicate that highly degradable and rapidly fermentable carbohydrates such as dietary inulin have a profound deworming effect on O. dentatum infection. PMID- 12885190 TI - Purification and characterization of a saccharide-binding protein from penetration glands of Diplostomum pseudospathaceum--a bifunctional molecule with cysteine protease activity. AB - A beta-1,3-glucan-binding lectin from the penetration glands of Diplostomum pseudospathaceum cercariae was isolated by affinity chromatography using yeast glucan and curdlan as affinity matrices. Further purification to homogeneity was performed by cation-exchange chromatography. The protein migrated as a double band around 24 kDa in gels after SDS-PAGE. The protein is of strongly basic nature--its pI shown by native IEF was around 10. The mass of the protein determined by MALDI-TOF mass spectrometry was 23.9 kDa. N-terminal sequence as well as some internal sequences showed significant alignments with several cysteine protease sequences found in databases. The protein bound a biotinylated synthetic analogue of the irreversible inhibitor of cysteine proteases, E-64 and, moreover, its proteolytic activity was demonstrated in substrate gels. The enzymatic activity could be inhibited by the cysteine protease inhibitor E-64; therefore, the investigated protein was considered to be a bifunctional molecule possessing both lectin and enzyme activities. Glycanohydrolytic activity was not proved. The detected characters of this molecule lead to a hypothesis on its role in penetration of Diplostomum cercariae into fish hosts--that of binding to the carbohydrates of fish mucus and concurrent cleaving of protein components of the mucus and skin. PMID- 12885191 TI - Population structure and recruitment of the ectoparasite Argulus coregoni Thorell (Crustacea: Branchiura) on a fish farm. AB - The population structure and recruitment of Argulus coregoni was monitored at a Finnish fish farm during the open water periods of 1999 and 2001 by weekly sampling of attached argulids. In 2001 the numbers of rainbow trout examined increased in the autumn when the A. coregoni population was declining. When the water temperature exceeded 10 degrees C, at the end of May, A. coregoni egg hatching commenced. A mean number of 98 (S.D. +/- 5.4) juvenile A. coregoni was recorded on each fish, before the start of female egg laying in July 1999. The abundance of lice was lower in 2001. The main recruitment of A. coregoni juveniles occurred in early summer, but the hatching of eggs continued until September. However, no pulses of hatching were recorded later in the summer and the numbers of lice on fish substantially decreased after mid-July in both years. On average, only 0.6 (S.D. +/- 0.72) parasites/fish were found between August and late October 2001 and none in November. We suggest that there is 1 main A. coregoni generation annually in Central Finland. Environmental conditions, especially temperature, affects the population cycle of A. coregoni: we found a more synchronous and intense population cycle during the summer 1999, when the early summer was warmer than in 2001. The overall sex ratio (female:male) of A. coregoni was nearly 1:1 in June 1999, but was male biased (1:1.4) in June 2001. It was also shown that from July onwards, many females detached from the fish host in order to lay their eggs on the bottom of the pond. Large males were often bigger than the largest females between mid-July and early September 2001, when as many as 3.7 (s.r. +/- 1.48) times more males than females were present in the lice samples. PMID- 12885192 TI - Anti-complement activity in the saliva of phlebotomine sand flies and other haematophagous insects. AB - The saliva of haematophagous insects has a series of pharmacological activities which may favour blood feeding. In the present study, an inhibitory effect on the complement system was observed in salivary extracts obtained from the phlebotomine sand flies Lutzomyia longipalpis and Lu. migonei. Saliva from Lu. longipalpis was capable of inhibiting both the classical and alternative pathways, while that from Lu. migonei acted only on the former. Other haematophagous insect species were screened for inhibition of the classical pathway. The triatomine bugs Panstrongylus megistus, Triatoma brasiliensis and Rhodnius prolixus were also able to inhibit the classical pathway whereas the mosquito Aedes aegyti and flea Ctenocephalides felis were not. The activity of Lu. longipalpis saliva on the classical pathway was partially characterized. The inhibitor is a protein of Mr 10000-30000 Da, which is very resistant to denaturation by heat. The inhibition of the complement system by phlebotomine sand flies may have a role in the transmission of Leishmania to the vertebrate hosts. The inhibitor molecule is thus a promising component of a vaccine to target salivary immunomodulators. PMID- 12885193 TI - Review of current technologies for urinary incontinence: strengths and limitations. AB - A wide range of continence products is available, and this paper focuses on products to prevent incontinence (such as urinals and commodes) and to contain or manage urinary incontinence (such as absorbent pads, penile sheaths and urethral catheters). Drawing on results from published clinical evaluations at the Continence Product Evaluation Network at UCL and at other centres, the strengths and limitations of the major categories of incontinence products currently on the market are reviewed. It is concluded that, although products for continence have improved considerably over the last 20 years, there is considerable scope for the designer and engineer to improve on current products. PMID- 12885194 TI - Engineered structures for use in disposable incontinence products. AB - The background and evolution of absorbent disposable hygiene products is discussed. The current technology used to produce disposable hygiene products such as baby diapers, feminine hygiene products and adult incontinence management products is shown to have a number of manufacturing and in-use problems. Alternative technologies, such as multilayer composite webs offer the opportunity to achieve better product weight control, and therefore better control over the other related properties, reduced raw material waste and improved functional characteristics. There are still problems to be resolved when using such composite materials, but the potential of these new technologies has now been established. PMID- 12885195 TI - Mapping liquid distribution in absorbent incontinence products. AB - This paper reviews methods available for mapping the distribution of fluid in incontinence pad materials to assist with evaluating existing products and developing new ones, and to provide data for building and validating predictive models. Specifically, the following technologies are considered and their strengths and limitations described: discrete sensors based on conductance, temperature or optical measurements, optical imaging, gravimetric methods, X-ray imaging and magnetic resonance imaging. It is suggested that the ideal method would enable fluid distribution to be mapped in three dimensions with good spatial and time resolution in single materials and composite structures of simple and complex geometries under static and dynamic mechanical loading. It would also allow liquid to be mapped in products when worn by users. It is concluded that, although each existing method meets some of these requirements, and each requirement is met, at least reasonably well, by at least one method, improved techniques are needed. The particular need for methods that can provide some measurement of liquid saturation within absorbent products, both in the laboratory and in real use, is highlighted. In many cases, simple methods used appropriately are sufficient to elicit the important aspects of liquid transport and storage within absorbent products. PMID- 12885196 TI - Experimental and theoretical study of the spread of fluid from a point source on an inclined incontinence bed-pad. AB - The spread of fluid from a localized source on to a flat fibrous sheet is studied. The sheet is inclined at an angle, alpha, to the horizontal, and the areal flux of the fluid released is Qa. A new experimental study is described where the dimensions of the wetted region are measured as a function of time t, Qa and alpha (>0). The down-slope length, Y, grows according to Y approximately (Qa t)(2/3) (sin alpha)(1/3); for high discharge rates and low angles of inclination, the cross-slope width, X, grows as approximately (Qa t)(1/2), while for low discharge rates or high angles of inclination, the cross-slope transport is dominated by infiltration and X approximately 2(2Ks psi* t)(1/2), where Ks is the saturated permeability and psi* is the characteristic value of capillary pressure. A scaling analysis of the underlying non-linear advection diffusion equation describing the infiltration process confirms many of the salient features of the flow observed. Good agreement is observed between the collapse of the numerical solutions and experimental results. The broader implications of these results for incontinence bed-pad research are briefly discussed. PMID- 12885197 TI - Wetting between parallel fibres; column-unduloid and column disintegration transitions. AB - Wetting phenomena are studied for a fibre bundle consisting of three parallel cylinders of radius b. Two types of long liquid column (a disintegrated and a channel-filling column) and a state akin to the unduloid can form, depending on whether the cylinder-cylinder separation, 2d, is larger or smaller than certain critical values. The dimensionless threshold values d/b have been calculated, when gravity can be neglected and the liquid-fibre contact angle is zero. The threshold value for the morphological transition between disintegrated columns and channel-filling columns occurs for (d/b)tr1 = 0.07869, and the transition between channel-filling column and unduloid sets in when (d/b)tr2 = 2.85337. The existence of the first type of morphological transition is validated by computer simulation based on the Ising model and the Monte Carlo method, as well as by experiments carried out with steel cylinders and epoxy resin. PMID- 12885198 TI - Polymer surface properties and their effect on the adhesion of Proteus mirabilis. AB - A problem encountered in patients undergoing long-term catheterization of the urinary tract is that of encrustation and blockage of the catheter by crystalline bacterial biofilms. This is principally caused by the action of the urease producing pathogen Proteus mirabilis. A major aim of this work is to develop materials resistant to encrustation. Here, the effects of polymer surface properties on the adhesion of P. mirabilis are examined. Spin-coated polymer films were characterized through contact angle measurements to give the Lifschitz van der Waals, electron acceptor and electron donor terms of the surface free energy, gamma(s)LW, gamma(s)+ and gamma(s)- respectively. A parallel-plate flow cell was used to assess adhesion to these polymer films of P. mirabilis suspended in an aqueous phosphate buffer, pH 7.4, ionic strength 0.26 mol/kg. P. mirabilis was found to adhere significantly less (p < 0.02) to films of agarose, poly(2 hydroxyethylmethacrylate) and cross-linked poly(vinyl alcohol) than to more hydrophobic materials. These polymer films were found to be strongly electron donating, i.e. possessing large gamma(s)-. Films examined using scanning electron microscopy mostly showed no evidence of roughness down to a scale of 1-10 microm. The better performance is thought to be due to a repulsive interaction with the bacterial surface caused by acid/base-type interactions. PMID- 12885199 TI - The further development of the active urine collection device: a novel continence management system. AB - Continence difficulties affect the lives of a substantial minority of the population. Women are far more likely than men to be affected by urinary incontinence but the range of management options for them is limited. There has been considerable interest in developing an external urine collection system for women but without success to date. This paper describes the development and preliminary clinical testing of an active urine collection device (AUCD), which could provide a solution for sufferers. The device uses stored vacuum, protected by a high bubble point filter, to remove urine as quickly as it is produced. This allows a small battery-operated pump to provide the required vacuum, enabling the device to be portable. Two different types of non-invasive patient/device interface were developed, and tested by volunteers: urinal and small pad. The slimline urinal was popular with users although liquid noise was a problem. The pad interface was successful on occasions but further work is necessary to produce a reliable pad. This study has successfully demonstrated that a prototype AUCD liquid handling system can remove urine at clinically relevant flowrates. While further development is required, volunteer tests have shown that the AUCD could be a useful advance in continence management. PMID- 12885200 TI - Developing alternative devices to the long-term urinary catheter for draining urine from the bladder. AB - The self-retaining urinary catheter is used for long-term drainage of urine from the bladder only as a last resort because of serious associated complications, yet it remains a routine method for managing older and disabled patients with loss of bladder control. Blockage of the catheter from calcified deposits within its lumen is a common occurrence, obstructing the passage of urine and causing an urgent, unpredictable problem for patients, carers and the nursing staff. The need for further research on the subject has been recognized for many years. The SuPort Project aimed to develop an alternative suprapubic urine collection system. This report outlines the approach adopted towards the design and selection of the novel device, the production problems that ensued and the small clinical trial of a modified prototype. PMID- 12885201 TI - A system for logging incontinence events using a simple disposable sensor. AB - Many elderly people entering residential or nursing care are already incontinent to some degree, relying on incontinence pads to deal with the consequences. A proportion of these people have been shown to exhibit a regular pattern in their incontinence, which opens up the possibility of mitigating the problem by instituting an individual toileting regime for the person. This can reduce their reliance on incontinence pads, both improving their quality of life, and reducing the cost of care. This paper covers the development and evaluation of a sensor for detecting incontinence events, suitable for use in this setting, and describes the design of an associated electronic logger. The devices form part of an assessment system intended to identify a pattern in incontinence where it exists, and to help with the design of the toilet regime for an individual. The requirement is that the system must reliably record incontinence events, and present the information describing them in a manner appropriate to the users of the devices, who are likely to be non-technical and non-specialist. PMID- 12885202 TI - Bladder reconstruction--from cells to materials. AB - Surgical reconstruction of the urinary bladder is performed on patients of all ages for a diverse range of conditions, including congenital abnormalities, bladder dysfunction, trauma and cancer. The most common material utilized to augment or replace the bladder during these procedures is a segment of the patient's own intestine. However, this procedure ('enterocytoplasty') is associated with significant clinical complications that arise due to the exposure of the epithelial lining of the intestine to urine. A number of alternative approaches are being actively developed to find a practical and functional substitute for native bladder tissue. These range from 'composite enterocystoplasty', where the de-epithelialized intestine wall is lined with bladder epithelial cells that have been propagated in vitro, to augmenting the urinary system with natural or synthetic biomaterials that may incorporate in vitro-propagated cells. However, if tissue-engineered products are to have therapeutic application in bladder reconstruction, a number of issues remain to be addressed; these issues are discussed briefly below. PMID- 12885203 TI - A new device for the treatment of female stress urinary incontinence. AB - Treatment for stress urinary incontinence (SUI) comprises a broad range of possible interventions. Non-surgical options include absorbent pads, vaginal weights and cones, biofeedback and minimally invasive techniques such as urethral bulking agents (UBAs). Surgical interventions range in complexity from sling surgery and suspension techniques to more major surgeries such as burch colposuspension. Each option has its challenges and limitations. This paper will focus on UBAs, which are implantable materials whose purpose is to augment urethral tissue function and restore continence. The characteristics required of such materials, and the challenges to be overcome when incorporating them in a successful product design, will be described and discussed. Particular attention will be given to the latest developments in the administration of polydimethylsiloxane elastomer UBA. PMID- 12885205 TI - Non-infectious and iatrogenic diseases of salmon in commercial aquaculture. AB - Based on current commercial aquaculture production practices, the production cycle for Atlantic salmon (Salmo salar) extends over a 36-month period during which time the fish are first exposed to intensive freshwater rearing conditions followed by transportation to marine netpen sites for ongrowing. It is predictable that, because of the duration of the production cycle and the variety of water conditions, deleterious environmental conditions have many opportunities to affect salmon health directly and indirectly. Furthermore, diseases which are iatrogenic arise from current methods used to prevent or treat infectious diseases. Specific, more frequently encountered examples are explored, with reference to the possible roles which these diseases may have in favouring the onset of infectious diseases. PMID- 12885204 TI - Canine distemper, a re-emerging morbillivirus with complex neuropathogenic mechanisms. AB - Paramyxoviruses are responsible for a wide variety of diseases both in humans and in animals. Common to many paramyxoviruses is the fact that they can cause neurological symptoms in their final host. Newly discovered paramyxoviruses, such as the Hendra and Nipah viruses, show the same pattern of pathogenesis as that of the paramyxoviruses already known. Canine distemper virus (CDV) is a well-studied member of the genus Morbillivirus. Study of the neuropathogenesis of CDV might give insight into disease mechanisms and suggest approaches for the prevention of other recently discovered paramyxovirus infections. PMID- 12885206 TI - American canine hepatozoonosis. AB - American canine hepatozoonosis is an emerging, tick-transmitted infection of domestic dogs caused by a recently recognized species of apicomplexan parasite, Hepatozoon americanum. The known definitive host of the protozoan is the Gulf Coast tick, Amblyomma maculatum. Presently recognized intermediate hosts include the domestic dog and the coyote, Canis latrans. Laboratory-reared larval or nymphal A. maculatum can be infected readily by feeding to repletion on a parasitemic intermediate host; sporogony requires 35-40 days. Transmission of infection to the dog has been produced experimentally by oral administration of mature oocysts or oocyst-containing ticks. Canine disease follows experimental exposure in 4-6 weeks and is characterized by systemic illness, extreme neutrophilic leukocytosis, muscle and bone pain, and proliferation of periosteal bone. Histopathological findings include multifocal skeletal and cardiac myositis associated with escape of mature merozoites from within the host-cell environment. There is also rapid onset of periosteal activation and osteogenesis and, less frequently, glomerulopathy and amyloidosis. Sequential stages of development of H. americanum in both the dog and the tick have been elucidated. Gamonts potentially infectious to ticks have been observed in peripheral blood leukocytes of the dog in as few as 28 days after exposure to oocysts. Young coyotes experimentally exposed to a canine strain of H. americanum acquired disease indistinguishable from that of similarly exposed young dogs. PMID- 12885207 TI - Global perspectives on nematode parasite control in ruminant livestock: the need to adopt alternatives to chemotherapy, with emphasis on biological control. AB - Effective, sustainable control of nematode parasites of grazing livestock is becoming evermore challenging and difficult. This is largely due to two contrasting issues. One is the rapid escalation of resistance to anthelmintic drugs, which is arguably the greatest problem now facing the small ruminant industries worldwide. Secondly, there is the increasing trend towards organic farming, in which there is prohibition of the prophylactic use of all chemical compounds. Livestock producers urgently need non-chemotherapeutic alternatives in parasite control. Researchers have responded to this challenge and a variety of quite different approaches have been the subject of intense investigation in many countries for several decades now. These vary in relation to their stage of development for on-farm use, their utility, and their applicability across the spectrum of grazing livestock enterprises throughout the world. One relatively recent innovation is the biological control approach to nematode parasites. This has now reached the stage of commercialization. This review focuses on these issues and provides an overview of the possible ways in which the biological control of nematode parasites could be employed in grazing ruminant livestock systems worldwide. PMID- 12885208 TI - A key mechanism of pathogenesis in sheep infected with the nematode Teladorsagia circumcincta. AB - Infection of sheep with the abomasal nematode Teladorsagia circumcincta can cause a relative protein deficiency and reduce growth rate in growing lambs. A key event appears to be the destruction of junctions between epithelial cells. If the infection is heavy or prolonged, this leads to increased mucus production, hyperplasia, decreased acid production, gastrinemia, inappetance and pepsinogenemia. The severity of the infection depends upon the extent of concurrent infection, the nutritional status of the host and genetically controlled variation in the ability to mount protective immune responses. PMID- 12885209 TI - Cryptosporidium and host resistance: historical perspective and some novel approaches. AB - Cryptosporidium parvum is recognized as a major cause of diarrheal disease in neonatal bovine calves. In addition, this protozoan parasite has emerged as an important cause of disease in both immunocompromised and immunocompetent humans. Despite years of research, no consistently effective means of prevention or treatment are readily available for cryptosporidiosis in any species. Infection through ingestion of contaminated water has been widely documented; C. parvum was reported to be responsible for the largest waterborne outbreak of infectious disease in US history. In addition to its role as a primary disease agent, C. parvum has potential to initiate or exacerbate other gastrointestinal disorders, such as inflammatory bowel disease. Thus, control of C. parvum infection in both animals and humans remains an important objective. Research in our laboratory has focused on understanding mechanisms of resistance to C. parvum. We have demonstrated that acquisition of intestinal flora increases resistance to C. parvum. Substances present in the intestinal mucosa of adult animals can transfer resistance when fed to susceptible infants. Both expression of intestinal enzymes and rate of proliferation of epithelial cells may be altered following C. parvum infection. These and other changes may have profound effects on host resistance to C. parvum. PMID- 12885210 TI - Lay TB testing: DEFRA issues its proposals. PMID- 12885211 TI - How do you measure quality of life? PMID- 12885212 TI - Early dissemination of foot-and-mouth disease virus through sheep marketing in February 2001. AB - The results of epidemiological investigations suggest that livestock on up to 79 premises, spread widely throughout the British Isles, may have been exposed to infection by foot-and-mouth disease (FMD) virus by the movement of infected sheep before the first case of the disease was confirmed at an abattoir in Essex on February 20, 2001. A further 36 premises may have been infected by this route before the national livestock movement ban was imposed on February 23. PMID- 12885213 TI - Comparison of two treatments for preventing dogs eating their own faeces. AB - Twenty-eight domestic dogs with the behavioural problem of eating their own faeces were treated in two ways. Half of them were treated with a citronella spray collar, and the others were treated with sound therapy. To assess the relative efficacy of the treatments their owners rated the severity of their dog's faeces eating for a week before the study began, during each of three weeks of treatment, and at the end of a fourth week during which they had not been treated. The owners reported a significantly lower incidence of the behavioural problem during the first week of both treatments, but in the dogs treated by sound therapy its incidence subsequently increased. The behavioural problem was reduced most effectively in the dogs treated with the spray collar, and continued to decrease during the period of treatment. PMID- 12885214 TI - Plasma and erythrocyte fatty acids in captive Asian (Elephas maximus) and African (Loxodonta africana) elephants. AB - The fatty acid components of the plasma triglycerides and the phospholipid fractions of the red blood cells of a captive group of two African (Loxodonta africana) and four Asian (Elephas maximus) elephants were investigated. All the animals received the same diet of hay, fruits and vegetables, and concentrates. A comparison with data from free-ranging African elephants or Asian work-camp elephants showed that the captive elephants had lower proportions of polyunsaturated fatty acids (PUFAs), and for several lipid fractions a higher n 6:n-3 ratio, than their counterparts in the wild or under the more natural, in terms of diet, work-camp conditions. The difference in PUFA content was smaller in the African than in the Asian elephants. The captive Asian elephants tended to have lower levels of n-3 and total unsaturated fatty acids in their red blood cells than the captive African elephants. PMID- 12885215 TI - Measurement of 25-hydroxycholecalciferol in captive grey parrots (Psittacus e erithacus). PMID- 12885216 TI - Clostridium tertium as a cause of enteritis in cattle. PMID- 12885217 TI - FMD control strategies. PMID- 12885218 TI - Dealing with exotic disease. PMID- 12885219 TI - Religious slaughter. PMID- 12885220 TI - Religious slaughter. PMID- 12885221 TI - Deer-related traffic accidents. PMID- 12885222 TI - Supply of POMs. PMID- 12885223 TI - Ethical issues in veterinary practice. PMID- 12885224 TI - Effects of photosynthetic reaction center H protein domain mutations on photosynthetic properties and reaction center assembly in Rhodobacter sphaeroides. AB - Purple bacterial photosynthetic reaction center (RC) H proteins comprise three cellular domains: an 11 amino acid N-terminal sequence on the periplasmic side of the inner membrane; a single transmembrane alpha-helix; and a large C-terminal, globular cytoplasmic domain. We studied the roles of these domains in Rhodobacter sphaeroides RC function and assembly, using a mutagenesis approach that included domain swapping with Blastochloris viridis RC H segments and a periplasmic domain deletion. All mutations that affected photosynthesis reduced the amount of the RC complex. The RC H periplasmic domain is shown to be involved in the accumulation of the RC H protein in the cell membrane, while the transmembrane domain has an additional role in RC complex assembly, perhaps through interactions with RC M. The RC H cytoplasmic domain also functions in RC complex assembly. There is a correlation between the amounts of membrane-associated RC H and RC L, whereas RC M is found in the cell membrane independently of RC H and RC L. Furthermore, substantial amounts of RC M and RC L are found in the soluble fraction of cells only when RC H is present in the membrane. We suggest that RC M provides a nucleus for RC complex assembly, and that a RC H/M/L assemblage results in a cytoplasmic pool of soluble RC M and RC L proteins to provide precursors for maximal production of the RC complex. PMID- 12885225 TI - Functional differences of photosystem II from Synechococcus elongatus and spinach characterized by flash induced oxygen evolution patterns. AB - Detailed comparative studies of flash induced oxygen evolution patterns in thylakoids from the thermophilic cyanobacterium Synechococcus elongatus (S. elongatus; also referred to as Thermosynechococcus elongatus) and from spinach led to the following results: (i) the miss parameter alpha of S. elongatus thylakoids exhibits a pronounced temperature dependence with a minimum of 7% at 25 degrees C and values of 17 and 10% at 3 and 35 degrees C, respectively, while for spinach thylakoids alpha decreases continuously from 18% at 35 degrees C down to 8% at 3 degrees C; (ii) at all temperatures, the double hit probability beta exceeds in S. elongatus the corresponding values of spinach by an increment Delta beta of about 3%; (iii) at 20 degrees C the slow relaxation of the oxidation states S(2) and S(3) is about 15 and 30 times, respectively, slower in S. elongatus than in spinach, while the reduction of these S states by tyrosine Y(D) is 2-3 times faster; (iv) the reaction S(0)Y(D)(ox) --> S(1)Y(D) is slower by a factor of 4 in S. elongatus as compared to spinach; and (v) the activation energies of S state dark relaxations in S. elongatus are all within a factor of 1.5 as compared to the previously reported values from spinach thylakoids [Vass, I., Deak, Z., and Hideg, E. (1990) Biochim. Biophys. Acta 1017, 63-69; Messinger, J., Schroder, W. P., and Renger, G. (1993) Biochemistry 32, 7658-7668], but the difference between the activation energies of the slow S(2) and S(3) decays is significantly larger in S. elongatus than in spinach. These results are discussed in terms of differences between cyanobacteria and higher plants on the acceptor side of PSII and a shift of the redox potential of the couple Y(D)/Y(D)(ox). The obtained data are also suitable to address questions about effects of the redox state of Y(D) on the miss probability and the possibility of an S state dependent miss parameter. PMID- 12885226 TI - Hydralysin, a novel animal group-selective paralytic and cytolytic protein from a noncnidocystic origin in hydra. AB - In Cnidaria, the production of neurotoxic polypeptides is attributed to the ectodermal stinging cells (cnidocytes), which are discharged for offensive (prey capture) and/or defensive purposes. In this study, a new paralysis-inducing (neurotoxic) protein from the green hydra Chlorohydra viridissima was purified, cloned, and expressed. This paralytic protein is unique in that it (1) is derived from a noncnidocystic origin, (2) reveals a clear animal group-selective toxicity, (3) possesses an uncommon primary structure, remindful of pore-forming toxins, and (4) has a fast cytotoxic effect on insect cells but not on the tested mammalian cells. The possible biological role of such a noncnidocystic toxin is discussed. PMID- 12885227 TI - Modeling domino effects in enzymes: molecular basis of the substrate specificity of the bacterial metallo-beta-lactamases IMP-1 and IMP-6. AB - Metallo-beta-lactamases can hydrolyze a broad spectrum of beta-lactam antibiotics and thus confer resistance to bacteria. For the Pseudomonas aeruginosa enzyme IMP 1, several variants have been reported. IMP-6 and IMP-1 differ by a single residue (glycine and serine at position 196, respectively), but have significantly different substrate spectra; while the catalytic efficiency toward the two cephalosporins cephalothin and cefotaxime is similar for both variants, IMP-1 is up to 10-fold more efficient than IMP-6 toward cephaloridine and ceftazidime. Interestingly, this biochemical effect is caused by a residue remote from the active site. The substrate-specific impact of residue 196 was studied by molecular dynamics simulations using a cationic dummy atom approach for the zinc ions. Substrates were docked in an intermediate structure near the transition state to the binding site of IMP-1 and IMP-6. At a simulation temperature of 100 K, most complexes were stable during 1 ns of simulation time. However, at higher temperatures, some complexes became unstable and the substrate changed to a nonactive conformation. To model stability, six molecular dynamics simulations at 100 K were carried out for all enzyme-substrate complexes. Stable structures were further heated to 200 and 300 K. By counting stable structures, we derived a stability ranking score which correlated with experimentally determined catalytic efficiency. The use of a stability score as an indicator of catalytic efficiency of metalloenzymes is novel, and the study of substrates in a near-transition state intermediate structure is superior to the modeling of Michaelis complexes. The remote effect of residue 196 can be described by a domino effect: upon replacement of serine with glycine, a hole is created and a stabilizing interaction between Ser196 and Lys33 disappears, rendering the neighboring residues more flexible; this increased flexibility is then transferred to the active site. PMID- 12885228 TI - Solution structure of a steroid-DNA complex with cholic acid residues sealing the termini of a Watson-Crick duplex. AB - The three-dimensional structure of a covalent hybrid between cholic acid and the self-complementary DNA hexamer 5'-TGCGCA-3' was solved via two-dimensional NMR and restrained torsion angle molecular dynamics. In the complex, refined to a pairwise rmsd of 0.64 A, the steroid binds to the terminal T:A base pairs via extensive van der Waals contacts but without any hydrogen bonds or detectable dipole-dipole interactions. The contacts involve the methyl groups as well as one edge of the streoid's sterane skeleton and both nucleobases and the deoxyriboses of the terminal base pair of the DNA. The surprising shape complementarity between steroid and the undisturbed DNA termini explains the increase in fidelity and affinity observed for hybridization probes bearing bile acid residues. Since the hydroxyl groups of the steroid do not contribute to the binding of the DNA, they may be derivatized, potentially giving access to a new class of specific binders for blunt ends of Watson-Crick duplexes. PMID- 12885229 TI - Solution structure and RNA interactions of the RNA recognition motif from eukaryotic translation initiation factor 4B. AB - Eukaryotic initiation factor 4B (eIF4B) is a multidomain protein with a range of activities that serves primarily to promote association of messenger RNA to the 40S ribosomal subunit during translation initiation. We report here the solution structure of the eIF4B RNA recognition motif (RRM) domain. It adopts a classical RRM fold, with a beta alpha beta beta alpha beta topology. The most striking difference with other RRM structures is in the disposition of loop 3, which connects the beta 2 and beta 3 strands and is implicated in RNA recognition. This loop folds down against the body of the RRM and exhibits restricted motion on a milli- to microsecond time scale. Although it contributes to a large basic patch on the RNA binding surface, it does not protrude out from the domain as observed in other RRM structures, possibly implying a different mode of RNA binding. On its own, the core RRM domain provides only a relative weak interaction with RNA targets and appears to require extensions at the N- and C-terminus for high affinity binding. PMID- 12885230 TI - Molecular dynamics simulations of pentapeptides at interfaces: salt bridge and cation-pi interactions. AB - Peptide-membrane interactions are important for understanding the binding, partitioning, and folding of membrane proteins; the activity of antimicrobial and fusion peptides; and a number of other processes. We describe molecular dynamics simulations (10-25 ns) of two pentapeptides Ace-WLXLL (with X = Arg or Lys side chain) (White, S. H., and Wimley, W.C. (1996) Nat. Struct. Biol. 3, 842-848) in water and three different membrane mimetic systems: (i) a water/cyclohexane interface, (ii) water-saturated octanol, and (iii) a solvated dioleoylphosphatidylcholine bilayer. A salt bridge is found between the protonated Arg or Lys side chains with the carboxyl terminus at the three interfaces. In water/cyclohexane, the salt bridge is most exposed to the water phase and least stable. In water/octanol and the lipid bilayer systems, the salt bridge once formed persists throughout the simulations. In the lipid bilayer, the salt bridge is more stable when the peptide penetrates deeper into the bilayer. In one of two peptides, a cation-pi interaction between the Arg and the Trp side chains is stable in the lipid bilayer for about 15 ns before breaking. In all cases, the conformations of the peptides are restricted by their presence at the interface and can be assigned to a few major conformational clusters. Side chains facing the water phase are most mobile. In the lipid bilayer, the peptides remain in the interface area, where they overlap with the carbonyl area of the lipid bilayer and perturb the local density profile of the bilayer. The tryptophan side chain remains in the water-lipid interface, where it interacts with the lipid choline group and forms hydrogen bonds with the ester carbonyl of the lipid and with water in the interface. PMID- 12885231 TI - Using motif-based methods in multiple genome analyses: a case study comparing orthologous mesophilic and thermophilic proteins. AB - Protein motifs represent highly conserved regions within protein families and are generally accepted to describe critical regions required for protein stability and/or function. In this comprehensive analysis, we present a robust, unique approach to identify and compare corresponding mesophilic and thermophilic sequence motifs between all orthologous proteins within 44 microbial genomes. Motif similarity is determined through global sequence alignment of mesophilic and thermophilic motif pairs, which are identified by a greedy algorithm. Our results reveal only modest correlation between motif and overall sequence similarity, highlighting the rationale of motif-based approaches in comprehensive multigenome comparisons. Conserved mutations reflect previously suggested physiochemical principles for conferring thermostability. Additionally, comparisons between corresponding mesophilic and thermophilic motif pairs provide key biochemical insights related to thermostability and can be used to test the evolutionary robustness of individual structural comparisons. We demonstrate the ability of our unique approach to provide key insights in two examples: the TATA box binding protein and glutamate dehydrogenase families. In the latter example, conserved mutations hint at novel origins leading to structural stability differences within the hexamer structures. Additionally, we present amino acid composition data and average protein length comparisons for all 44 microbial genomes. PMID- 12885232 TI - Structural and dynamic properties of the HIV-1 tat transduction domain in the free and heparin-bound states. AB - An 11-residue basic domain of the HIV-1 tat protein, termed the tat transduction domain (TTD), has been shown to mediate transfer of biomolecules across biological membranes. The mechanism of TTD-mediated membrane translocation is currently unknown but thought to involve binding to heparan sulfate, which is found in proteoglycans that are ubiquitously present on cell surfaces. To study the mechanism of TTD-mediated membrane translocation, the TTD was fused to the C terminus of a model cargo protein, the IgG binding domain of streptococcal protein G (PG) to form PG-TTD. NMR studies of PG-TTD in the free state indicated that the structure of the PG moiety of PG-TTD was not perturbed by the presence of the TTD and that the TTD moiety is in an extended conformation. Heteronuclear relaxation measurements of PG-TTD in the free state show that the TTD moiety of PG-TTD is relatively mobile (e.g., the average S(2) value of the TTD and PG core are approximately 0.54 and approximately 0.84, respectively). PG-TTD has been shown to bind to heparin by isothermal titration calorimetry (K(D) = 0.37 microM, Delta H = -12 kcal/mol, Delta S = -11 cal/mol/T). NMR spectroscopy demonstrated that heparin binds to the TTD moiety of PG-TTD. The heteronuclear relaxation measurements of PG-TTD in complex with heparin show that the TTD becomes less dynamic when bound to heparin (average S(2) value of the TTD is 0.69 in the presence of heparin). A model for the first step of TTD-mediated entry into cells is presented. PMID- 12885233 TI - Is the V3 loop involved in HIV binding to CD4? AB - The entry of the human immunodeficiency virus into cells requires the interaction of the viral envelope glycoprotein gp120 with CD4 and a chemokine receptor. The gp120 binding site has been previously mapped to the Ig-CDR2-like region of CD4 first domain. A second area of this domain (Ig-CDR3-like region) is involved in gp120-CD4 interactions, but its gp120 counterpart remained so far unknown. Using a photoaffinity labeling experiment, we demonstrate that a peptide, mapping the (307-330)m region of HIV-MN-gp120 V3 loop, binds a sequence including a part of the Ig-CDR3-like region. These results may contribute to explain the complex mechanism of human immunodeficiency virus penetration, helping the development of new therapeutic agents. PMID- 12885234 TI - Design of weakly basic thrombin inhibitors incorporating novel P1 binding functions: molecular and X-ray crystallographic studies. AB - To prepare weakly basic thrombin inhibitors with modified S1 anchoring groups, two series of compounds were synthesized by reaction of guanidine or aminoguanidine with acyl halides and N,N-disubstituted carbamoyl chlorides. pK(a) measurements of these acylated guanidines/aminoguanidines showed a reduced basicity, with pK(a) values in the range of 8.4-8.7. These molecules typically showed inhibition constants in the range of 150-425 nM against thrombin and 360 965 nM against trypsin, even though some bulky derivatives, such as N,N diphenylcarbamoylguanidine/aminoguanidine and their congeners, showed much stronger thrombin inhibitory activity, with inhibition constants in the range of 24-42 nM. Unexpectedly, very long incubation times with both proteases revealed that aminoguanidine derivatives behaved as irreversible inhibitors. To assess the molecular basis responsible for the high affinity observed for these molecules toward thrombin, the crystal structure of the thrombin-hirugen-N,N diphenylcarbamoylaminoguanidine complex has been solved at 1.90 A resolution. The structural analysis of the complex revealed an unexpected interaction mode with the protease, resulting in an N,N-diphenylcarbamoyl intermediate covalently bound to the catalytic serine as a consequence of its hydrolysis together with the release of the aminoguanidine moiety. Surprisingly, in this covalent adduct a phenyl group was found in the S1 specificity pocket, which usually recognizes positively charged residues. These findings provide new insights in the design of low basicity serine protease inhibitors. PMID- 12885235 TI - Fibrillar aggregates of the tumor suppressor p53 core domain. AB - Alzheimer's disease, Parkinson's disease, cystic fibrosis, prion diseases, and many types of cancer are considered to be protein conformation diseases. Most of them are also known as amyloidogenic diseases due to the occurrence of pathological accumulation of insoluble aggregates with fibrillar conformation. Some neuroblastomas, carcinomas, and myelomas show an abnormal accumulation of the wild-type tumor suppressor protein p53 either in the cytoplasm or in the nucleus of the cell. Here we show that the wild-type p53 core domain (p53C) can form fibrillar aggregates after mild perturbation. Gentle denaturation of p53C by pressure induces fibrillar aggregates, as shown by electron and atomic force microscopies, by binding of thioflavin T, and by circular dichroism. On the other hand, heat denaturation produced granular-shaped aggregates. Annular aggregates similar to those found in the early aggregation stages of alpha-synuclein and amyloid-beta were also observed by atomic force microscopy immediately after pressure treatment. Annular and fibrillar aggregates of p53C were toxic to cells, as shown by MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] reduction assay. Interestingly, the hot-spot mutant R248Q underwent similar aggregation behavior when perturbed by pressure or high temperature. Fibrillar aggregates of p53C contribute to the loss of function of p53 and seed the accumulation of conformationally altered protein in some cancerous cells. PMID- 12885236 TI - Deletion of DnaK's lid strengthens binding to the nucleotide exchange factor, GrpE: a kinetic and thermodynamic analysis. AB - In this study, we have used surface plasmon resonance (SPR) and isothermal microtitration calorimetry (ITC) to study the mechanism of complex formation between the Hsp70 molecular chaperone, DnaK, and its cochaperone, GrpE, which is a nucleotide exchange factor. Experiments were geared toward understanding the influence of DnaK's three domains, the ATPase (residues 1-388), substrate-binding (residues 393-507), and lid (residues 508-638) domains, on complex formation with GrpE. We show that the equilibrium dissociation constants for the interaction of GrpE with wtDnaK, lidless DnaK(2-517), the ATPase domain (2-388), and the substrate-binding fragment (393-507) are 64 (+/-16) nM, 4.0 (+/-1.5) nM, 35 (+/ 10) nM, and 67 (+/-11) microM, respectively, and that the on-rate constant for the different reactions varies by over 4 orders of magnitude. SPR experiments revealed that GrpE-DnaK(393-507) complex formation is inhibited by added peptide and abolished when the 33-residue flexible "tail" of GrpE is deleted. Such results strongly suggest that the 33-residue flexible N-terminal tail of GrpE binds in the substrate-binding pocket of DnaK. This unique mode of binding between GrpE's tail and DnaK contributes to, but does not fully explain, the decrease in K(d) from 64 to 4 nM upon deletion of DnaK's lid. The possibility that deletion of DnaK's lid creates a more symmetrically shaped molecule, with enhanced affinity to GrpE, is also discussed. Our results reveal a complex set of molecular interactions between DnaK and its cochaperone GrpE. We discuss the impact of each domain on complex formation and dissociation. PMID- 12885237 TI - Amino acid determinants in cyclooxygenase-2 oxygenation of the endocannabinoid anandamide. AB - The endocannabinoid arachidonylethanolamide (AEA, anandamide) is an endogenous ligand for the cannabinoid receptors and has been shown to be oxygenated by cyclooxygenase-2 (COX-2). We examined the structural requirements for COX mediated, AEA oxygenation using a number of substrate analogues and site-directed mutants of COX-2. Fourteen AEA analogues were synthesized and tested as COX substrates. These studies identified the hydroxyl moiety of AEA as a critical determinant in the ability of COX enzymes to effect robust endocannabinoid oxygenation. In addition, these studies suggest that subtle structural modifications of AEA analogues near the ethanolamide moiety can result in pronounced changes in their ability to serve as COX-2 substrates. Site-directed mutagenesis studies have permitted the development of a model of AEA binding within the COX-2 active site. As with arachidonic acid, the omega-terminus of AEA binds in a hydrophobic alcove near the top of the COX-2 active site. The polar ethanolamide moiety of AEA, like the carboxylate of arachidonate, interacts with Arg-120 at the bottom of the COX-2 active site. Mutation of Tyr-385 prevents AEA oxygenation, suggesting that, as in the case of other COX substrates, AEA metabolism is initiated by Tyr-385-mediated hydrogen abstraction. Thus, AEA binds within the COX-2 active site in a conformation roughly similar to that of arachidonic acid. However, important differences have been identified that account for the isoform selectivity of AEA oxygenation. Importantly, the COX-2 side pocket and Arg-513 in particular are critical determinants of the ability of COX-2 to efficiently generate prostaglandin H(2) ethanolamide. The reduced efficiency of COX-1-mediated, AEA oxygenation can thus be explained by the absence of an arginine residue at position 513 in this isoform. Mutational analysis of Leu-531, an amino acid located directly across from the COX-2 side pocket, suggests that AEA is shifted away from this hydrophobic residue and toward Arg-513 relative to arachidonic acid. Coupled with earlier observations with the endocannabinoid 2-arachidonylglycerol, these results indicate that one possible function of the highly conserved COX-2 active site side pocket is to promote endocannabinoid oxygenation. PMID- 12885238 TI - Thermodynamic linkage in the GrpE nucleotide exchange factor, a molecular thermosensor. AB - GrpE is the nucleotide exchange factor for the Escherichia coli molecular chaperone DnaK, the bacterial homologue of Hsp70. In the temperature range of the bacterial heat shock response, the long helices of GrpE undergo a helix-to-coil transition, and GrpE exhibits non-Arrhenius behavior with respect to its nucleotide exchange function. It is hypothesized that GrpE acts as a thermosensor and that unwinding of the long helices of E. coli GrpE reduces its activity as a nucleotide exchange factor. In turn, it was proposed that temperature-dependent down-regulation of the activity of GrpE may increase the time in which DnaK binds its substrates at higher temperatures. A combination of thermodynamic and hydrodynamic techniques, in concert with the luciferase refolding assay, were used to characterize a molecular mechanism in which the long helices of GrpE are thermodynamically linked with the beta-domains via an intramolecular contact between Phe86 and Arg183. These "thermosensing" long helices were found to be necessary for full activity as a nucleotide exchange factor in the luciferase refolding assay. Point mutations in the beta-domains and in the long helices of GrpE destabilized the beta-domains. Engineered disulfide bonds in the long helices alternately stabilized the long helices and the four-helix bundle. This allowed the previously reported 75 degrees C thermal transition seen in the excess heat capacity function as monitored by differential scanning calorimetry to be further characterized. The observed thermal transition represents the unfolding of the four-helix bundle and the beta-domains. The thermal transitions for these two domains are superimposed but are not thermodynamically linked. PMID- 12885239 TI - Engineering the primary substrate specificity of Streptomyces griseus trypsin. AB - Streptomyces griseus trypsin (SGT) was chosen as a model scaffold for the development of serine proteases with enhanced substrate specificity. Recombinant SGT has been produced in a Bacillus subtilis expression system in a soluble active form and purified to homogeneity. The recombinant and native proteases have nearly identical enzymatic properties and structures. Four SGT mutants with alterations in the S1 substrate binding pocket (T190A, T190P, T190S, and T190V) were also expressed. The T190P mutant demonstrated the largest shift to a preference for Arg versus Lys in the P1 site. This was shown by a minor reduction in catalytic activity toward an Arg-containing substrate (k(cat) reduction of 25%). The crystal structures of the recombinant SGT and the T190P mutant in a complex with the inhibitor benzamidine were obtained at high resolution (approximately 1.9 A). The increase in P1 specificity, achieved with minimal effect on the catalytic efficiency, demonstrates that the T190P mutant is an ideal candidate for the design of additional substrate specificity engineered into the S2 to S4 binding pockets. PMID- 12885240 TI - Structural basis for the quinone reduction in the bc1 complex: a comparative analysis of crystal structures of mitochondrial cytochrome bc1 with bound substrate and inhibitors at the Qi site. AB - Cytochrome bc(1) is an integral membrane protein complex essential to cellular respiration and photosynthesis. The Q cycle reaction mechanism of bc(1) postulates a separated quinone reduction (Q(i)) and quinol oxidation (Q(o)) site. In a complete catalytic cycle, a quinone molecule at the Q(i) site receives two electrons from the b(H) heme and two protons from the negative side of the membrane; this process is specifically inhibited by antimycin A and NQNO. The structures of bovine mitochondrial bc(1) in the presence or absence of bound substrate ubiquinone and with either the bound antimycin A(1) or NQNO were determined and refined. A ubiquinone with its first two isoprenoid repeats and an antimycin A(1) were identified in the Q(i) pocket of the substrate and inhibitor bound structures, respectively; the NQNO, on the other hand, was identified in both Q(i) and Q(o) pockets in the inhibitor complex. The two inhibitors occupied different portions of the Q(i) pocket and competed with substrate for binding. In the Q(o) pocket, the NQNO behaves similarly to stigmatellin, inducing an iron sulfur protein conformational arrest. Extensive binding interactions and conformational adjustments of residues lining the Q(i) pocket provide a structural basis for the high affinity binding of antimycin A and for phenotypes of inhibitor resistance. A two-water-mediated ubiquinone protonation mechanism is proposed involving three Q(i) site residues His(201), Lys(227), and Asp(228). PMID- 12885241 TI - High-resolution solution structure of the beryllofluoride-activated NtrC receiver domain. AB - Bacterial receiver domains mediate the cellular response to environmental changes through conformational changes induced by phosphorylation of a conserved aspartate residue. While the structures of several activated receiver domains have recently been determined, there is substantial variation in the conformational changes occurring upon activation. Here we present the high resolution structure of the activated NtrC receiver domain (BeF(3)(-)-NtrC(r) complex) determined using NMR data, including residual dipolar couplings, yielding a family of structures with a backbone rmsd of 0.57 +/- 0.08 A, which is compared with the previous lower-resolution structure of the phosphorylated protein. Both phosphorylation and beryllofluoride addition induce a shift in register and an axial rotation of alpha-helix 4. In this high-resolution structure, we are able to observe a concerted change in the positions of Thr82 and Tyr101; this correlated change in two conserved residues (termed Y-T coupling) has been considered a general feature of the conformational change in receiver domains upon activation. In NtrC, this correlated side chain shift, leading to the helix reorientation, is distinctly different from the smaller reorganization seen in other activated receiver domains, and involves numerous other residues which do not participate in conformational changes seen in the other systems. Titration of the activated receiver domain with peptides from the NtrC ATPase domain provides direct evidence for interactions on the rearranged face of the receiver domain, which are likely to be responsible for enabling assembly into the active aggregate. Analysis of the active structure also suggests that His84 may play a role in controlling the phosphate hydrolysis rate. PMID- 12885242 TI - Conservation in the CYP51 family. Role of the B' helix/BC loop and helices F and G in enzymatic function. AB - CYP51 (sterol 14 alpha-demethylase) is an essential enzyme in sterol biosynthetic pathways and the only P450 gene family having catalytically identical orthologues in different biological kingdoms. The proteins have low sequence similarity across phyla, and the whole family contains about 40 completely conserved amino acid residues. Fifteen of these residues lie in the secondary structural elements predicted to form potential substrate recognition sites within the P450 structural fold. The role of 10 of these residues, in the B' helix/BC loop, helices F and G, has been studied by site-directed mutagenesis using as a template the soluble sterol 14 alpha-demethylase of known structure, CYP51 from Mycobacterium tuberculosis (MT) and the human orthologue. Single amino acid substitutions of seven residues (Y76, F83, G84, D90, L172, G175, and R194) result in loss of the ability of the mutant MTCYP51 to metabolize lanosterol. Residual activity of D195A is very low, V87A is not expressed as a P450, and A197G has almost 1 order of magnitude increased activity. After purification, all of the mutants show normal spectral properties, heme incorporation, and the ability to be reduced enzymatically and to interact with azole inhibitors. Profound influence on the catalytic activity correlates well with the spectral response to substrate binding, effect of substrate stabilization on the reduced state of the P450, and substrate-enhanced efficiency of enzymatic reduction. Mutagenesis of corresponding residues in human CYP51 implies that the conserved amino acids might be essential for the evolutionary conservation of sterol 14 alpha demethylation from bacteria to mammals. PMID- 12885243 TI - Localization of a substrate binding site on the FeMo-cofactor in nitrogenase: trapping propargyl alcohol with an alpha-70-substituted MoFe protein. AB - Substitution of the MoFe protein alpha-70(Val) residue with Ala or Gly expands the substrate range of nitrogenase, allowing the reduction of larger alkynes, including propargyl alcohol (HC[triple bond]CCH(2)OH). Herein, we report characterization of the alpha-70(Val)(-->)(Ala) MoFe protein with propargyl alcohol trapped at the active site. The alpha-70(Ala) variant MoFe protein was rapidly frozen during reduction of propargyl alcohol, resulting in the conversion of the resting-state FeMo-cofactor EPR signal (S = 3/2 and g = [4.41, 3.60, 2.00]) to a new state (S = 1/2 and g = [2.123, 1.998, 1.986]). This EPR signal of the new state increased in intensity with increasing propargyl alcohol concentration, consistent with the binding of a single substrate. The EPR signal of the propargyl alcohol state showed temperature and microwave power dependencies markedly different from those of the classic FeMo-cofactor EPR signal, consistent with the difference in spin. The new state is analogous to that induced by the binding of the inhibitor CO ("lo CO" state) to FeMo-cofactor in the wild-type MoFe protein. The (13)C ENDOR spectrum of the alpha-70(Ala) MoFe protein with trapped (13)C-labeled propargyl alcohol exhibited three well resolved (13)C doublets centered at the (13)C Larmor frequency with isotropic hyperfine couplings of approximately 3.2, approximately 1.4, and approximately 0.7 MHz, indicating that the alcohol (or a fragment) is coordinated to the cofactor. The results presented here localize the binding site of propargyl alcohol to one [4Fe-4S] face of FeMo-cofactor and indicate roles for the alpha 70(Val) residue in controlling FeMo-cofactor reactivity. PMID- 12885244 TI - Three-dimensional model for meta-II rhodopsin, an activated G-protein-coupled receptor. AB - A novel approach that iteratively combined the results of energy calculations and experimental data was used to generate a three-dimensional (3D) model of the photoactivated state (R*) of bovine rhodopsin (Rh). The approach started with simplified energy calculations in an effort to find a set of sterically and energetically reasonable options for transmembrane (TM) helix arrangements with all-trans-retinal. Various 3D models of TM helix packing found by computations were then compared to limited site-directed spin-label experimental data regarding the transition of the TM helices of Rh in the inactive state (R) to those in the R* state to identify the most plausible model of the TM helical bundle. At the next step, all non-TM structural elements, such as the non-TM helix 8, the N- and C-terminal fragments, and the loops connecting TM helices, were reconstructed, and after the entire R* structure had been relaxed, all other currently available additional experimental data, both mutational and spectroscopic, on the structure of the meta-II state of rhodopsin were used to validate the resulting 3D model. PMID- 12885245 TI - Metal-1,4-dithio-2,3-dihydroxybutane chelates: novel inhibitors of the Rho transcription termination factor. AB - Rho is an enzyme that is essential for the growth and survival of Escherichia coli, and bicyclomycin (1) is its only known selective inhibitor. We show that metal (Cd(2+), Ni(2+), and Zn(2+)) complexes of 1,4-dithio-2,3-dihydroxybutanes (2) serve as effective and potent rho inhibitors with I(50) values that can exceed that of 1. Maximal inhibition for ZnCl(2) and L-dithiothreitol (2a) corresponded to Zn(2):L-DTT stoichiometry. The I(50) value for the 2:1 Zn-L-DTT solution was 20 microM, which made it 3 times more potent than 1 (I(50) = 60 microM). Kinetic studies showed that a Zn-L-DTT solution functioned as a noncompetitive inhibitor with respect to ATP in the rho poly(C)-dependent ATPase assay and as a competitive inhibitor with respect to ribo(C)(10) in the poly(dC).ribo(C)(10)-stimulated ATPase assay. These findings demonstrated that both 1 and a Zn-L-DTT solution disrupted rho-mediated ATP hydrolysis but that they inhibit using different mechanisms. Substitution of L-DTT with 1,2 ethanedithiol in ZnCl(2) solutions led to a comparable loss of rho poly(C) dependent ATPase activity, indicating that other metal chelates can serve as efficient inhibitors. The site and pathway of rho inhibition by the putative metal-1,4-dithio-2,3-dihydroxybutane chelates are discussed in light of the current data. PMID- 12885246 TI - Two redox-active beta-carotene molecules in photosystem II. AB - Photosystem II (PS II) contains secondary electron-transfer paths involving cytochrome b(559) (Cyt b(559)), chlorophyll (Chl), and beta-carotene (Car) that are active under conditions when oxygen evolution is blocked such as in inhibited samples or at low temperature. Intermediates of the secondary electron-transfer pathways of PS II core complexes from Synechocystis PCC 6803 and Synechococcus sp. and spinach PS II membranes have been investigated using low temperature near IR spectroscopy and electron paramagnetic resonance (EPR) spectroscopy. We present evidence that two spectroscopically distinct redox-active carotenoids are formed upon low-temperature illumination. The Car(+) near-IR absorption peak varies in wavelength and width as a function of illumination temperature. Also, the rate of decay during dark incubation of the Car(+) peak varies as a function of wavelength. Factor analysis indicates that there are two spectral forms of Car(+) (Car(A)(+) has an absorbance maximum of 982 nm, and Car(B)(+) has an absorbance maximum of 1027 nm) that decay at different rates. In Synechocystis PS II, we observe a shift of the Car(+) peak to shorter wavelength when oxidized tyrosine D (Y(D)*) is present in the sample that is explained by an electrostatic interaction between Y(D)* and a nearby beta-carotene that disfavors oxidation of Car(B). The sequence of electron-transfer reactions in the secondary electron transfer pathways of PS II is discussed in terms of a hole-hopping mechanism to attain the equilibrated state of the charge separation at low temperatures. PMID- 12885247 TI - A putative consensus sequence for the nucleotide-binding site of annexin A6. AB - Reaction-induced infrared difference spectroscopy (RIDS) has been used to investigate the nature of interactions of human annexin A6 (ANXA6) with nucleotides. RIDS results for ANXA6, obtained after the photorelease of GTP-gamma S, ATP, or P(i) from the respective caged compounds, were identical, suggesting that the interactions between the nucleotide and ANXA6 were dominated by the phosphate groups. Phosphate-induced structural changes in ANXA6 were small and affected only seven or eight amino acid residues. The GTP fluorescent analogue, 2'(3')-O-(2,4,6-trinitrophenyl)guanosine 5'-triphosphate (TNP-GTP), quenched tryptophan fluorescence of ANXA6 when bound to the protein. A binding stoichiometry of 1 mol of nucleotide/mol ANXA6 was established with a K(D) value of 2.8 microM for TNP-GTP. The bands observed on RIDS of ANXA6 halves (e.g., N terminal half, ANXA6a, and C-terminal half, ANXA6b) were similar to those of the whole molecule. However, their amplitudes were smaller by a factor of 2 compared to those of whole ANXA6. TNP-GTP bound to both fragments of ANXA6 with a stoichiometry of 0.5 mol/mol. However, the binding affinities of ANXA6a and ANXA6b differed from that of ANXA6. Simulated molecular modeling revealed a nucleotide-binding site which was distributed in two distinct domains. Residues K296, Y297, K598, and K644 of ANXA6 were less than 3 A from the bound phosphate groups of either GTP or ATP. The presence of two identical sequences in ANXA6 with the F-X-X-K-Y-D/E-K-S-L motif, located in the middle of ANXA6, at residues 293-301 (within ANXA6a) and at 641-649 (within ANXA6b), suggested that the F-X-X K-Y-D/E-K-S-L motif was the putative sequence in ANXA6 for nucleotide binding. PMID- 12885248 TI - Functional asymmetry of the human Na+/glucose transporter (hSGLT1) in bacterial membrane vesicles. AB - The functional characteristics of the forward and reverse transport modes of the human Na(+)/glucose transporter (hSGLT1) were investigated using plasma membrane vesicles of E. coli expressing the recombinant transporter. Correctly and inverse oriented vesicles were employed to measure the initial rates of methyl-alpha-D glucose uptake, under zero-trans conditions, as a function of Na(+), sugar, and phlorizin concentrations and membrane potential. This approach enabled the analysis of the two faces of hSGLT1 in parallel, revealing the reversibility of Na(+)/sugar cotransport. While the key characteristics of secondary active sugar transport were maintained in both modes, namely, Na(+) and voltage dependence, the kinetic properties of the two sides indicated a functional asymmetry of the transporter. That is, the apparent affinity for sugar and driver cation Na(+) exhibited a difference of more than 1 order of magnitude between the two modes. Furthermore, the selectivity pattern of ligands and the interaction of the transporter with the competitive inhibitor phlorizin were different. Whereas the high-affinity substrates, D-glucose and D-galactose, inhibited uptake of radioactive sugar tracer at their physiological concentrations (10 mM) in the forward reaction, they were poor inhibitors even at high concentrations in the reverse transport mode. Taken together, these results confirm the successful employment of E. coli to express and characterize a human membrane protein (hSGLT1), elucidating the functional asymmetry of this cotransporter. PMID- 12885249 TI - Characterization of a lactose permease mutant that binds IIAGlc in the absence of ligand. AB - Enzyme IIA(Glc) of the Escherichia coli phosphoenolpyruvate:glucose phosphotransferase system plays a direct role in regulating inducible transport systems. Dephosphorylated IIA(Glc) binds directly to lactose permease in a reaction that requires binding of a galactosidic substrate. A double-Cys mutation (Ile129 --> Cys/Lys131 --> Cys) was introduced into helix IV of the permease near the IIA(Glc) binding site in cytoplasmic loop IV/V and in the vicinity of the galactoside binding site at the interface of helices IV, V, and VIII. The mutant no longer requires galactoside for IIA(Glc) binding as demonstrated by both a [(125)I]IIA(Glc) binding assay and a newly developed fluorescence anisotropy assay. Further characterization of the mutant shows that it binds substrate with high affinity, but is almost completely defective in all modes of translocation across the cytoplasmic membrane. The data are consistent with the interpretation that the double mutant is locked in an inward-facing conformation. PMID- 12885250 TI - Addition of lysines to the 50/20 kDa junction of myosin strengthens weak binding to actin without affecting the maximum ATPase activity. AB - Much interest has centered on two surface loops in the motor domain to explain the differences in enzymatic and mechanical properties of myosin isoforms. We showed that two invariant lysines at the C-terminal end of loop 2, which is part of the actin-binding interface, are required to obtain actin activation [Joel et al. (2001) J. Biol. Chem. 276, 2998-3003]. Here we investigate the effects of increasing positive charge in the variable portion of loop 2 of smooth muscle heavy meromyosin (smHMM). Increasing the net positive charge by +4 increased the affinity for actin in the presence and absence of ATP. The K(m) for actin activated ATPase activity decreased 15-fold, but V(max) was unchanged, showing that "weak binding" of myosin for actin can be significantly strengthened without increasing the rate-limiting step for V(max). The mutant HMM had slower rates of in vitro motility and ADP release compared to WT HMM. ADP release and motility, which were both salt-dependent, correlated linearly with each other. Loop 2 thus plays a major role in setting the affinity for actin but also affects ADP release and motility. Because the actin- and nucleotide-binding regions communicate, mutations to one region can impact multiple facets of myosin's mechanical and enzymatic properties. PMID- 12885251 TI - Lipid-protein interactions and effect of local anesthetics in acetylcholine receptor-rich membranes from Torpedo marmorata electric organ. AB - The selectivity of lipid-protein interaction for spin-labeled phospholipids and gangliosides in nicotinic acetylcholine receptor-rich membranes from Torpedo marmorata has been studied by ESR spectroscopy. The association constants of the spin-labeled lipids (relative to phosphatidylcholine) at pH 8.0 are in the order cardiolipin (5.1) approximately equal to stearic acid (4.9) approximately equal to phosphatidylinositol (4.7) > phosphatidylserine (2.7) > phosphatidylglycerol (1.7) > G(D1b) approximately equal to G(M1) approximately equal to G(M2) approximately equal to G(M3) approximately equal to phosphatidylcholine (1.0) > phosphatidylethanolamine (0.5). No selectivity for mono- or disialogangliosides is found over that for phosphatidylcholine. Aminated local anesthetics were found to compete with spin-labeled phosphatidylinositol, but to a much lesser extent with spin-labeled stearic acid, for sites on the intramembranous surface of the protein. The relative association constant of phosphatidylinositol was reduced in the presence of the different local anesthetics to the following extents: tetracaine (55%) > procaine (35%) approximately benzocaine (30%). For stearic acid, only tetracaine gave an appreciable reduction (30%) in association constant. These displacements represent an intrinsic difference in affinity of the local anesthetics for the lipid-protein interface because the membrane partition coefficients are in the order benzocaine >> tetracaine approximately procaine. PMID- 12885252 TI - Differential interaction of GRK2 with members of the G alpha q family. AB - Regulators of G protein signaling (RGS) proteins bind to active G alpha subunits and accelerate the rate of GTP hydrolysis and/or block interaction with effector molecules, thereby decreasing signal duration and strength. RGS proteins are defined by the presence of a conserved 120-residue region termed the RGS domain. Recently, it was shown that the G protein-coupled receptor kinase 2 (GRK2) contains an RGS domain that binds to the active form of G alpha(q). Here, the ability of GRK2 to interact with other members of the G alpha(q) family, G alpha(11), G alpha(14), and G alpha(16), was tested. The signaling of all members of the G alpha(q) family, with the exception of G alpha(16), was inhibited by GRK2. Immunoprecipitation of full-length GRK2 or pull down of GST-GRK2-(45-178) resulted in the detection of G alpha(q), but not G alpha(16), in an activation dependent manner. Moreover, activated G alpha(16) failed to promote plasma membrane (PM) recruitment of a GRK2-(45-178)-GFP fusion protein. Assays with chimeric G alpha(q)(-)(16) subunits indicated that the C-terminus of G alpha(q) mediates binding to GRK2. Despite showing no interaction with GRK2, G alpha(16) does interact with RGS2, in both inositol phosphate and PM recruitment assays. Thus, GRK2 is the first identified RGS protein that discriminates between members of the G alpha(q) family, while another RGS protein, RGS2, binds to both G alpha(q) and G alpha(16). PMID- 12885253 TI - Protein transduction domains of HIV-1 and SIV TAT interact with charged lipid vesicles. Binding mechanism and thermodynamic analysis. AB - Cell-penetrating peptides (CPPs) traverse cell membranes of cultured cells very efficiently by a mechanism not yet identified. Recent theories for the translocation suggest either the binding of the CPPs to extracellular glycosaminoglycans or the formation of inverted micelles with negatively charged lipids. In the present study, the binding of the protein transduction domains (PTD) of human (HIV-1) and simian immunodeficiency virus (SIV) TAT peptide (amino acid residues 47-57, electric charge z(p) = +8) to membranes containing various proportions of negatively charged lipid (POPG) is characterized. Monolayer expansion measurements demonstrate that TAT-PTD insertion between lipids requires loosely packed monolayer films. For densely packed monolayers (pi > 29 mN/m) and lipid bilayers, no insertion is possible, and binding occurs via electrostatic adsorption to the membrane surface. Light scattering experiments show an aggregation of anionic lipid vesicles when the electric surface charge is neutralized by TAT-PTD, the observed stoichiometry being close to the theoretical value of 1:8. Membrane binding was quantitated with isothermal titration calorimetry and three further methods. The reaction enthalpy is Delta H degrees approximately equal to -1.5 kcal/mol peptide and is almost temperature independent with Delta C(p) degrees approximately 0 kcal/(mol K), indicating equal contributions of polar and hydrophobic interactions to the reaction heat capacity. The binding of TAT-PTD to the anionic membrane is described by an electrostatic attraction/chemical partition model. The electrostatic attraction energy, calculated with the Gouy-Chapman theory, accounts for approximately 80% of the binding energy. The overall binding constant, K(app), is approximately 10(3)-10(4) M(-1). The intrinsic binding constant (K(p)), corrected for electrostatic effects and describing the partitioning of the peptide between the lipid-water interface and the membrane, is small and is K(p) approximately 1-10 M(-1). Deuterium and phosphorus-31 nuclear magnetic resonance demonstrate that the lipid bilayer remains intact upon TAT-PTD binding. The NMR data provide no evidence for nonbilayer structures and also not for domain formation. This is further supported by the absence of dye efflux from single-walled lipid vesicles. The electrostatic interaction between TAT-PTD and anionic phosphatidylglycerol is strong enough to induce a change in the headgroup conformation of the anionic lipid, indicating a short-lived but distinct correlation between the TAT-PTD and the anionic lipids on the membrane outside. TAT-PTD has a much lower affinity for lipid membranes than for glycosaminoglycans, making the latter interaction a more probable pathway for CPP binding to biological membranes. PMID- 12885254 TI - Adhesion-associated and PKC-modulated changes in serine/threonine phosphorylation of p120-catenin. AB - p120-catenin (p120) was originally identified as a tyrosine kinase substrate, and subsequently shown to regulate cadherin-mediated cell-cell adhesion. Binding of the p120 Arm domain to E-cadherin appears to be necessary to maintain adequate cadherin levels for strong adhesion. In contrast, the sequence amino-terminal to the Arm domain confers a negative regulatory function that is likely to be modulated by phosphorylation. Several agents that induce rapid changes in cell cell adhesion, including PDBu, histamine, thrombin, and LPA, result in significant changes in p120 S/T phosphorylation. In some cases, these changes are PKC-dependent, but the relationship among adhesion, PKC activation, and p120 phosphorylation is unclear, in part because the relevant p120 phosphorylation sites are unknown. As a crucial step toward directly identifying the function of these modifications in adhesion, we have used two-dimensional tryptic mapping and site-directed mutagenesis to pinpoint the constitutive and PKC-modulated sites of p120 S/T phosphorylation. Of eight sites that have been identified, two were selectively phosphorylated in vitro by GSK3 beta, but in vivo treatment of cells with GSK3 beta inhibitors did not eliminate these sites. PKC stimulation in vivo induced potent dephosphorylation at S268, and partial dephosphorylation of several additional sites. Surprisingly, PKC also strongly induced phosphorylation at S873. These data directly link PKC activation to specific changes in p120 phosphorylation, and identify the target sites associated with the mechanism of PKC-dependent adhesive changes induced by agents such as histamine and PDBu. PMID- 12885255 TI - Electric field effects on the chlorophylls, pheophytins, and beta-carotenes in the reaction center of photosystem II. AB - We present an electric field modulated absorption spectroscopy (Stark effect) study of isolated photosystem II reaction center complexes, including a preparation in which the inactive pheophytin H(B) was exchanged for 13(1)-deoxo 13(1)-hydroxy-pheophytin. The results reveal that the Stark spectrum of the Q(x) and Q(y) transitions of the pheophytins has a second-derivative line shape, indicating that the Stark effect is dominated by differences in the dipole moment between the ground and the electronically excited states of these transitions (Delta mu). The Delta mu values for the Q(x) and Q(y) transitions of H(B) are small (Delta mu = 0.6-1.0 D f(-1)), whereas that of the Q(x) transition of the active pheophytin H(A) is remarkably large (Delta mu = 3 D f(-1)). The Stark spectrum of the red-most absorbing pigments also shows a second-derivative line shape, but this spectrum is considerably red-shifted as compared to the second derivative of the absorption spectrum. This situation is unusual but has been observed before in heterodimer special pair mutants of purple bacterial reaction centers [Moore, L. J., Zhou, H., and Boxer, S. G. (1999) Biochemistry 38, 11949 11960]. The red-shifted Stark spectra can be explained by a mixing of exciton states with a charge-transfer state of about equal energy. We conclude that the charge transfer state involves H(A) and its immediate chlorophyll neighbor (B(A)), and we suggest that this (B(A)(delta+)H(A)(delta-)) charge transfer state plays a crucial role in the primary charge separation reaction in photosystem II. In contrast to most other carotenes, the two beta-carotene molecules of the photosystem II reaction center display a very small Delta mu, which can most easily be explained by excitonic coupling of both molecules. These results favor a model that locates both beta-carotene molecules at the same side of the complex. PMID- 12885256 TI - Control of metalloprotein reduction potential: compensation phenomena in the reduction thermodynamics of blue copper proteins. AB - The reduction thermodynamics (Delta H degrees '(rc) and Delta S degrees '(rc)) for native Paracoccus versutus amicyanin, for Alcaligenes faecalis S-6 pseudoazurin, and for the G45P, M64E, and K27C variants of Pseudomonas aeruginosa azurin were measured electrochemically. Comparison with the data available for other native and mutated blue copper proteins indicates that the features of metal coordination and the electrostatic potential due to the protein matrix and the solvent control the reduction enthalpy in a straightforward way. However, the effects on the reduction potential are rather unpredictable owing to the entropic contribution to E degrees ', which is mainly determined by solvent reorganization effects. Analysis of all the Delta H degrees '(rc) and Delta S degrees '(rc) values available for this protein class indicates that enthalpy-entropy compensation occurs in the reduction thermodynamics of wt cupredoxins from different sources, as well as for mutants of the same species. The findings indicate that the reduction enthalpies and entropies for these species are strongly affected by reduction-induced reorganization of solvent molecules within the solvation sphere of the protein. The absence of a perfect enthalpy-entropy compensation is due to the fact that while the differences between reduction entropies are dominated by solvent reorganization effects, those between reduction enthalpies are significantly controlled by intrinsic molecular factors related to the selective stabilization of the reduced form by coordination features of the copper site and electrostatic effects at the interface with the protein matrix. PMID- 12885257 TI - Bipyrimidine photoproducts rather than oxidative lesions are the main type of DNA damage involved in the genotoxic effect of solar UVA radiation. AB - Exposure to solar UV radiation gives rise to mutations that may lead to skin cancer. UVA (320-340 nm) constitutes the large majority of solar UV radiation but is less effective than UVB (290-320 nm) at damaging DNA. Although UVA has been implicated in photocarcinogenesis, its contribution to sunlight mutagenesis has not been elucidated, and DNA damage produced by UVA remains poorly characterized. We employed HPLC-MS/MS and alkaline agarose gel electrophoresis in conjunction with the use of specific DNA repair proteins to determine the distribution of the various classes and types of DNA lesions, including bipyrimidine photoproducts, in Chinese hamster ovary cells exposed to pure UVA radiation, as well as UVB and simulated sunlight (lambda > 295 nm) for comparison. At UVA doses compatible with human exposure, oxidative DNA lesions are not the major type of damage induced by UVA. Indeed, single-strand breaks, oxidized pyrimidines, oxidized purines (essentially 8-oxo-7,8-dihydroguanine), and cyclobutane pyrimidine dimers (CPDs) are formed in a 1:1:3:10 ratio. In addition, we demonstrate that, in contrast to UVB and sunlight, UVA generates CPDs with a large predominance of TT CPDs, which strongly suggests that they are formed via a photosensitized triplet energy transfer. Moreover, UVA induces neither (6-4) photoproducts nor their Dewar isomers via direct absorption. We also show that UVA photons contained in sunlight, rather than UVB, are implicated in the photoisomerization of (6-4) photoproducts, a quickly repaired damage, into poorly repaired and highly mutagenic Dewar photoproducts. Altogether, our data shed new light on the deleterious effect of UVA. PMID- 12885258 TI - Quantitative identification of the protonation state of histidines in vitro and in vivo. AB - The C[bond]N coupling constants centered at the C(epsilon 1) and C(delta 2) carbons in histidine residues depend on the protonation state and tautomeric form of the imidazole ring, making them excellent indicators of pH or pK(a), and the ratio of the tautomeric states. In this paper, we demonstrate that the intensity ratios for the C(epsilon 1)-H and C(delta 2)-H cross-peaks measured with a constant time HSQC experiment without and with J(C[bond]N) amplitude modulation are determined by the ratios of the protonated and deprotonated forms and tautomeric states. This allows one to investigate the tautomeric state of histidines as well as their pK(a) in situations where changing the pH value by titration is difficult, for example, for in-cell NMR experiments. We apply this technique to the investigation of the bacterial protein NmerA and determine that the intracellular pH in the Escherichia coli cytoplasm is 7.1 +/- 0.1. PMID- 12885259 TI - Dynamic, thermodynamic, and kinetic basis for recognition and transformation of DNA by human immunodeficiency virus type 1 integrase. AB - Specific interactions between retroviral integrase (IN) and long terminal repeats are required for insertion of viral DNA into the host genome. To characterize quantitatively the determinants of substrate specificity, we used a method based on a stepwise increase in ligand complexity. This allowed an estimation of the relative contributions of each nucleotide from oligonucleotides to the total affinity for IN. The interaction of HIV-1 integrase with specific (containing sequences from the LTR) or nonspecific oligonucleotides was analyzed using a thermodynamic model. Integrase interacted with oligonucleotides through a superposition of weak contacts with their bases, and more importantly, with the internucleotide phosphate groups. All these structural components contributed in a combined way to the free energy of binding with the major contribution made by the conserved 3'-terminal GT, and after its removal, by the CA dinucleotide. In contrast to nonspecific oligonucleotides that inhibited the reaction catalyzed by IN, specific oligonucleotides enhanced the activity, probably owing to the effect of sequence-specific ligands on the dynamic equilibrium between the oligomeric forms of IN. However, after preactivation of IN by incubation with Mn(2+), the specific oligonucleotides were also able to inhibit the processing reaction. We found that nonspecific interactions of IN with DNA provide approximately 8 orders of magnitude in the affinity (Delta G degrees approximately equal to -10.3 kcal/mol), while the relative contribution of specific nucleotides of the substrate corresponds to approximately 1.5 orders of magnitude (Delta G degrees approximately equal to - 2.0 kcal/mol). Formation of the Michaelis complex between IN and specific DNA cannot by itself account for the major contribution of enzyme specificity, which lies in the k(cat) term; the rate is increased by more than 5 orders of magnitude upon transition from nonspecific to specific oligonucleotides. PMID- 12885262 TI - Bone as an effect compartment : models for uptake and release of drugs. AB - "Bone-seeking agents" are drugs characterised by high affinity for bone, and are disposed in bone for prolonged periods of time while maintaining remarkably low systemic concentrations. As a consequence, the bone becomes a reservoir for bone seeking agents, and a site of both desirable and adverse effects, depending on the pharmacological activities of the specific agent. For some agents, significant systemic effects may also be produced following their prolonged release from bone, a process that is governed mostly by the rate of bone remodelling. This review covers the pharmacokinetic and pharmacodynamic features of bone-seeking agents with different pharmacological properties, including drugs (bisphosphonates, drug-bisphosphonate conjugates, radiopharmaceuticals and fluoride), bone markers (tetracycline, bone imaging agents) and toxins (lead, chromium, aluminium). In addition, drugs that do not possess bone-seeking properties but are used for therapy of bone diseases (such as antibacterials for treatment of osteomyelitis) are discussed, along with targeting of these drugs to the bone by conjugation to bone-seeking agents, local delivery systems, and other approaches. The pharmacokinetic and pharmacodynamic behaviour of bone-seeking agents is extremely complex due to heterogeneity in bone morphology and physiology. This complexity, accompanied by difficulties in human bone research caused by ethical and other limitations, gave rise to modelling approaches to study bone drug disposition. This review describes the pharmacokinetic models that have been proposed to describe the pharmacokinetic behaviour of bone-seeking agents and predict bone concentrations of these agents for different doses and patient populations. Models of different types (compartmental and physiologically based) and of different complexity have been applied, but their relevance to drug effects in the bone tissue is limited since they describe the behaviour of the "average" drug molecule. Understanding of the cellular and molecular processes responsible for the heterogeneity of bone tissue will provide better comprehension of the influence of microenvironment on drug bone disposition and the resulting pharmacological response. PMID- 12885264 TI - Clinical pharmacokinetics of antiplatelet agents used in the secondary prevention of stroke. AB - Stroke is one of the leading causes of death and debilitation. Several million stroke survivors are alive throughout the world today. Prevention of recurrent stroke is of major importance to stroke survivors. Several pharmacological agents are currently available for use in secondary stroke prevention.Clopidogrel, the combination of immediate-release aspirin and extended-release dipyridamole and aspirin alone are the most widely recommended agents for use in the secondary prevention of strokes. Clopidogrel has shown superiority over aspirin in the combined endpoints of stroke, death and myocardial infarction. The immediate release aspirin/extended-release dipyridamole combination has shown superiority to aspirin alone in the secondary prevention of stroke. Dipyridamole has been studied as an antiplatelet agent for several decades. Early trials to prove its efficacy compared with aspirin were not favourable, and patients often experienced many adverse effects. Researchers began developing an extended release formulation in an effort to maintain therapeutic blood concentrations with less frequent daily administration and better adverse effect profile. Pharmacokinetic analysis of this new product showed it to have a more consistent and reproducible absorption compared with immediate-release dipyridamole. The rate of absorption of extended-release dipyridamole is considerably slower than that of immediate-release dipyridamole, while similar plasma concentrations are maintained to optimise antiplatelet efficacy. This allows extended-release dipyridamole to be administered twice daily rather than four times daily.A large scale randomised trial was conducted with extended-release dipyridamole 200mg in combination with immediate-release aspirin 25mg given twice daily. The combination product showed a greater efficacy at preventing a recurring stroke then either agent administered alone. Indirect comparisons with clopidogrel show that the combination of immediate-release aspirin/extended-release dipyridamole may be more effective than clopidogrel at preventing a recurring stroke. PMID- 12885266 TI - Microbial pathogen genomes - new strategies for identifying therapeutic and vaccine targets. AB - Efficient mining of genomic sequence information from multiple pathogens for therapeutic and vaccine targets requires efficient tools. Fortunately, robust methods applicable to whole genomes have been developed and applied in the past few years to identify genes essential for growth or virulence and to detect potential vaccine targets. Successful approaches to identify potential therapeutic targets include a variety of ingenious uses of nearly random transposon insertions, more directed methods such as antisense and insertion duplication mutagenesis, and expression profiling facilitated by microarrays. Vaccine targets have been identified by gene fusion and expression experiments to discover gene products that are immunogenic in humans or animal models. All genome-wide methods require focused secondary assays to validate the findings, but these genomic methods excel at reducing to a manageable number the genes to be examined further. This editorial reviews the latest developments in genome wide target identification tools. PMID- 12885265 TI - Pharmacokinetics of tazarotene cream 0.1% after a single dose and after repeat topical applications at clinical or exaggerated application rates in patients with acne vulgaris or photodamaged skin. AB - OBJECTIVE: To evaluate the safety and pharmacokinetics of tazarotene cream 0.1% under standard (face only) or exaggerated (15% body surface area, including the face) application conditions after a single dose and after repeat topical applications once daily to patients with acne vulgaris or photodamaged skin. METHODS: Two separate, randomised, single-centre, nonblinded, parallel-group pharmacokinetic studies were conducted. In one study, tazarotene cream 0.1% was applied either to the face of eight female patients with moderate acne or to 15% body surface area of ten female patients with severe acne. In the other study, tazarotene cream 0.1% was applied either to the face (six females, two males) or to 15% body surface area (8 females, 8 males) of patients with photodamaged skin. In both studies, tazarotene cream 0.1% was applied once daily (except on days 1 and 2) for 30 days. Blood was drawn for measurement of plasma concentrations of tazarotenic acid at defined time intervals after application of the cream. Plasma tazarotenic acid concentrations were determined by a validated gas chromatography tandem mass spectrometry method with a lower limit of quantification of 0.005 microg/L. RESULTS: At exaggerated application rates in patients with acne vulgaris, the maximum average peak concentration (C(max)) and 24-hour area under the concentration-time curve (AUC) values of tazarotenic acid were (mean +/- SD) 1.20 +/- 0.41 microg/L (n = 10) and 17.0 +/- 6.1 microg. h/L (n = 10), respectively, and occurred on day 15. The single highest C(max) was 1.91 microg/L. At standard application rates in patients with acne vulgaris, the maximum average C(max) and AUC values of tazarotenic acid were 0.10 +/- 0.06 microg/L (n = 8) and 1.54 +/- 1.01 microg. h/L (n = 8), respectively, and occurred on day 15. At exaggerated application rates in patients with photodamaged skin, the maximum average C(max) and AUC values of tazarotenic acid were (mean +/- SD) 1.75 +/- 0.53 microg/L (n = 16) and 23.8 +/- 7.0 microg. h/L (n = 16), respectively, and occurred on day 22. The single highest C(max) was 3.43 microg/L on day 29. At standard application rates in patients with photodamaged skin, the maximum average C(max) and AUC values of tazarotenic acid were 0.236 +/- 0.255 microg/L (n = 8) and 2.44 +/- 1.38 microg. h/L (n = 8), respectively, and occurred on day 15. Gender had no influence on the systemic exposure of tazarotenic acid. The most common treatment-related adverse events were signs and symptoms of local irritation, of mild or moderate severity. CONCLUSIONS: The pharmacokinetics of tazarotene cream 0.1% in patients with acne vulgaris or photodamaged skin are similar. The maximum average plasma concentrations of tazarotenic acid after topical application of tazarotene cream 0.1% to the face were less than 0.25 microg/L. The maximum average plasma concentrations of tazarotenic acid following application to an exaggerated body surface area (15%) were less than 1.8 microg/L. PMID- 12885263 TI - Whole body pharmacokinetic models. AB - The aim of the current review is to summarise the present status of physiologically based pharmacokinetic (PBPK) modelling and its applications in drug research, and thus serve as a reference point to people interested in the methodology. The review is structured into three major sections. The first discusses the existing methodologies and techniques of PBPK model development. The second describes some of the most interesting PBPK model implementations published. The final section is devoted to a discussion of the current limitations and the possible future developments of the PBPK modelling approach. The current review is focused on papers dealing with the pharmacokinetics and/or toxicokinetics of medicinal compounds; references discussing PBPK models of environmental compounds are mentioned only if they represent considerable methodological developments or reveal interesting interpretations and/or applications.The major conclusion of the review is that, despite its significant potential, PBPK modelling has not seen the development and implementation it deserves, especially in the drug discovery, research and development processes. The main reason for this is that the successful development and implementation of a PBPK model is seen to require the investment of significant experience, effort, time and resources. Yet, a substantial body of PBPK-related research has been accumulated that can facilitate the PBPK modelling and implementation process. What is probably lagging behind is the expertise component, where the demand for appropriately qualified staff far outreaches availability. PMID- 12885267 TI - Regulators of G protein signalling: potential targets for treatment of allergic inflammatory diseases such as asthma. AB - Asthma, a disease that affects nearly 15% of the world's population, is characterised by lung inflammation and reversible airway obstruction, which leads to wheezing and dyspnoea. Asthma is a prototype for allergic processes initiated by tissue inflammatory leukocytes, such as mast cells, whose secreted mediators recruit lymphocytes and eosinophils to the lung parenchyma. Signals transmitted through G-protein-coupled receptors (GPCRs) contribute to both the development and perpetuation of allergic processes, and pharmacological agents that block or stimulate GPCR action have been a mainstay of allergic disease therapy. Despite the widespread use of GPCR-targeted agents, little is understood about intracellular regulation of G protein pathways in immune cells. Regulators of G protein signalling (RGS proteins) enhance G protein deactivation and may contribute to the specificity and precision characteristic of GPCR signalling pathways. This review discusses the emerging functions of RGS proteins in immune processes and inflammatory states such as asthma, and their potential value as therapeutic targets for the treatment of allergic disease. PMID- 12885268 TI - ADAM33: a novel therapeutic target for asthma. AB - The incidence of asthma worldwide is increasing, and the disease has a large unmet clinical need. Despite the availability of anti-inflammatory and bronchodilator medication, there is persisting morbidity and mortality. New approaches are needed to understand the role that structural changes in the airways (remodelling) play in this process. Studies of the genetic basis of asthma have identified the ADAM33 (a disintegrase and metalloproteinase 33) gene, a novel member of the ADAM family of zinc-dependent metalloproteases, as a risk factor for the development of asthma and bronchial hyperresponsiveness (BHR). The identification of ADAM33 as a major risk factor involved in the pathogenesis of BHR and airway wall remodelling provides insight into the pathogenesis of asthma and represents a novel therapeutic target. PMID- 12885269 TI - Melanin-concentrating hormone functions in the nervous system: food intake and stress. AB - Melanin-concentrating hormone (MCH) is a cyclic neuropeptide, which centrally regulates food intake and stress. MCH induces food intake in rodents and, more generally, acts as an anabolic signal in energy regulation. In addition, MCH seems to be activatory on the stress axis. Two receptors for MCH in humans have very recently been characterised, namely, MCH-R1 and MCH-R2. MCH-R1 has received considerable attention, as potent and selective antagonists acting at that receptor display anxiolytic, antidepressant and/or anorectic properties. Feeding and affective disorders are both debilitating conditions that have become serious worldwide health threats. There are as yet no efficient and/or safe cures that could contain the near-pandemia phenomen of both diseases. Thus, the discovery of MCH-R1 antagonists may lead to the development of valuable drugs to treat obesity, anxiety and depressive syndromes. In addition, it opens wide avenues to probe additional functions of the peptide, both in the brain and in the peripheral nervous system. PMID- 12885270 TI - Contributions of P2X3 homomeric and heteromeric channels to acute and chronic pain. AB - ATP acts as a fast neurotransmitter by activating a family of ligand-gated ion channels, the P2X receptors. Functional homomeric (P2X(3)) and heteromeric (P2X(2/3)) receptors are highly localised on primary sensory afferent neurons that transmit nociceptive sensory information. Activation of these P2X(3)containing channels may provide a specific mechanism whereby ATP, released via synaptic transmission or by cellular injury, elicits pain. The physiological relevance of the pro-nociceptive actions of ATP is supported by data demonstrating that the exogenous peripheral or spinal administration of ATP and other P2X receptor agonists elicits nociceptive behaviour and increases sensitivity to noxious stimuli in both humans and laboratory animals. The nociceptive effects of P2X receptor agonists are also enhanced in the presence of inflammatory mediators. Both permanent (P2X(3) gene knockout) and transient (P2X(3) antisense) receptor gene disruption studies in laboratory rodents have provided hypoalgesic phenotypes, further supporting a role for P2X(3) subunits in contributing to the expression of pain. More recently, the acute systemic administration of a highly selective non-nucleotide P2X(3) antagonist, A317491, has been shown to fully block specific types of chronic inflammatory and neuropathic pain in animal models in the absence of cardiovascular and CNS side effects associated with other analgesic compounds. Therefore, both genetic and pharmacological approaches have provided converging evidence that activation of P2X(3)-containing channels is an important mediator of persistent nociceptive signalling. The available data also indicate potential discrete roles for homomeric P2X(3) and heteromeric P2X(2/3) receptor activation in acute and chronic pain. PMID- 12885271 TI - Gremlin - a putative pathogenic player in progressive renal disease. AB - Progressive renal fibrosis is the end process of renal injury leading to kidney failure. Current therapies for chronic renal failure aim to slow this process but fail to halt its progression. As the mechanisms involved in glomerulosclerosis and tubulointerstitial fibrosis are unravelled, potential treatments for this growing clinical problem should emerge. Gremlin, a developmental regulator of bone morphogenetic proteins (BMPs), has recently been implicated in processes such as glomerulosclerosis, tubulointerstitial fibrosis and cellular hypertrophy, and may represent a novel therapeutic target in progressive renal diseases. PMID- 12885272 TI - Molecular targets in the inhibition of angiogenesis. AB - Angiogenesis, the process of blood vessel formation, is crucial for malignant tumour growth and metastases; therefore, it has become an attractive target for anticancer therapy. Theoretically applicable to most solid tumours, this therapy may be advantageous over existing cytotoxic therapy, since it is directed at genetically stable endothelium growing within tumours rather than at malignant cells, which acquire resistance to treatment. Many promising angiogenesis inhibitors have been developed, although their activity has yet to be demonstrated in human clinical trials. To improve therapeutic benefit, this may require further insight into tumour angiogenesis, development of appropriate surrogate markers of activity, treatment of early stage neoplastic disease and probably a combination of different classes of antiangiogenesis agents to overcome redundant mechanisms of angiogenesis control. PMID- 12885273 TI - Molecular targets as therapeutic strategies in the management of breast cancer. AB - Although the molecular and genetic determinants of most sporadic breast cancers remain unknown, increasing understanding of molecular and genetic events affecting breast carcinogenesis has provided information about the potential roles of specific biomarkers in tumour development and spread. It is now recognised that mutations of some tumour suppressor genes appear to play important early roles in the formation of some breast cancers. In addition, alterations in proto-oncogenes may contribute to the development of some breast cancers. The study of breast tumour biology at the molecular level has led to the development of targeted drug design, which provides a variety of agents targeted at specific molecules for the prevention, diagnosis and treatment of breast cancer. This review will describe the recognised molecular targets in breast cancer. PMID- 12885275 TI - Reducing inhaled corticosteroids in asthma is just the start. PMID- 12885276 TI - Rhabdomyolysis secondary to interaction of fusidic acid and simvastatin. PMID- 12885277 TI - Screening for genital Chlamydia trachomatis infection: are men the forgotten reservoir? PMID- 12885278 TI - Helical computed tomography for lung cancer screening. PMID- 12885279 TI - The protection of human genetic information. PMID- 12885274 TI - Cellular FLICE-inhibitory protein: an attractive therapeutic target? AB - Cellular FLIP (c-FLIP), also known as FLICE-inhibitory protein, has been identified as an inhibitor of apoptosis triggered by engagement of death receptors (DRs) such as Fas or TRAIL (TNF-related apoptosis-inducing ligand). cFLIP is recruited to DR signalling complexes, where it prevents caspase activation. Animal models have indicated that c-FLIP plays an important role in T cell proliferation and heart development. Abnormal c-FLIP expression has been identified in various diseases such as multiple sclerosis (MS), Alzheimer's disease (AD), diabetes mellitus, rheumatoid arthritis (RA) and various cancers. This review focuses on recent insights into c-FLIP dysregulation associated with human diseases and addresses the possibilities of using c-FLIP as a therapeutic target. PMID- 12885280 TI - Disability in older Australians: projections for 2006-2031. AB - OBJECTIVES: To provide detailed projections for the prevalence of disability and associated common health conditions for older Australians for the period 2006 2031. DESIGN: Secondary analyses of datasets (national 1998 Survey of Disability, Ageing and Carers; and projections of Australia's population from 2006-2031) collected by the Australian Bureau of Statistics. OUTCOME MEASURES: (i) The projected number of people with differing levels of disability (core activity restrictions in self-care, mobility or communication) up to 2031; (ii) The projected number of people with the main health conditions associated with disability in 2006 and 2031. RESULTS: Projections indicate a 70% increase in the number of older people with profound disability over the next 30 years. The main conditions associated with profound or severe core activity restriction in older Australians are musculoskeletal, nervous system, circulatory and respiratory conditions and stroke. CONCLUSIONS: In the future, there will be many more older Australians requiring assistance because of disability. This will present a challenge to families, friends, volunteers and paid service providers. The Australian planning ratio for residential aged-care services and community aged care services should be changed to take account of the shift to an older population with greater need of support. PMID- 12885281 TI - Untreated hypertension among Australian adults: the 1999-2000 Australian Diabetes, Obesity and Lifestyle Study (AusDiab). AB - OBJECTIVE: To measure the prevalence of untreated hypertension in Australian adults, and examine the associations with clinical and lifestyle factors. DESIGN: AusDiab, a cross-sectional survey conducted between May 1999 and December 2000, involved participants from 42 randomly selected census districts throughout Australia. PARTICIPANTS: Of 20 347 eligible people aged >or= 25 years who completed a household interview, 11 247 attended a physical examination (response rate, 55%). MAIN OUTCOME MEASURES: The prevalence of hypertension (blood pressure >or= 140/90 mmHg or self-reported use of antihypertensive drugs) and its treatment; associations of clinical and lifestyle factors with the treatment of hypertension; and adequacy of treatment for primary and secondary prevention of cardiovascular disease. RESULTS: The prevalence of hypertension was 28.6 per 100 (95% CI, 25.0-32.3), and the prevalence of untreated hypertension was 15.2 per 100 (95% CI, 13.2-17.2). Of those with untreated hypertension, 80.8% (95% CI, 74.7%-85.0%) had had a blood pressure check within the preceding 12 months. At least one modifiable lifestyle factor was present in 71.7% (95% CI, 68.5%-74.8%) of participants with untreated hypertension. Although lower risk clinical characteristics of younger age and lack of hyperlipidaemia were independently associated with untreated hypertension, 53.5% warranted treatment based on current cardiovascular disease prevention guidelines and multivariable absolute risk assessment. CONCLUSIONS: Considerable scope remains for reducing the burden of cardiovascular disease through lifestyle modification and rational treatment of hypertension. PMID- 12885282 TI - Mobile telephone use among Melbourne drivers: a preventable exposure to injury risk. AB - OBJECTIVE: To determine the rate of handheld mobile telephone use among motor vehicle drivers. DESIGN AND SETTING: Observational study of motor vehicle drivers at three times (10: 00-11: 00; 14: 00-15: 00; 17: 00-18: 00) on three consecutive Fridays in October 2002 at 12 highway sites in metropolitan Melbourne. MAIN OUTCOME MEASURES: Rates of mobile phone use overall and by sex and age group, highway site (major metropolitan road, central business district, freeway exit ramp) and time of day (morning, afternoon, evening). RESULTS: 315 of 17 023 drivers were observed using mobile phones (18.5 users/1000 drivers; 95% CI, 16.5 20.6). Men had a slightly higher rate of use (19.0; 95% CI, 16.5-21.6) than women (17.5; 95% CI, 14.1-20.9), but the difference was not significant. Older drivers (50 years or more) had a significantly lower rate (4.8; 95% CI, 2.5-7.0) than middle-aged (21.9; 95% CI, 18.8-25.1) or young drivers (23.2; 95% CI, 18.9-27.5). Central business district drivers had a slightly, but not significantly, higher rate (20.5; 95% CI, 16.8-24.3) compared with those on major metropolitan roads (16.7; 95% CI, 13.3-20.2) or freeway exit ramps (18.2; 95% CI, 14.8-21.6). The rate of mobile phone use was significantly higher in the evening (23.5; 95% CI, 19.8-27.3) compared with the morning (16.0; 95% CI, 12.6-19.4) and afternoon (15.2; 95% CI, 11.9-18.4). CONCLUSION: Mobile phone use is common among Melbourne metropolitan drivers despite restrictive legislation. This issue needs to be further addressed by Victoria Police and public health and education agencies. Similar research is indicated to determine the extent of mobile phone use in other states. PMID- 12885283 TI - Cardiovascular risk among urban Aboriginal people. AB - OBJECTIVE: To describe the results of a program for detecting high cardiovascular risk in an urban Aboriginal community. DESIGN: Cardiovascular risk assessment program conducted between January 1998 and October 1999. Participants completed a questionnaire and underwent a physical assessment and biochemical tests. PARTICIPANTS: 738 self-selected members of the Perth Aboriginal community (332 men, 406 women; age range, 18-79 years). RESULTS: The participants represented approximately a fifth of the Perth Aboriginal population aged 25-64 years (those aged 18-24 years comprised < 5% of Aboriginals aged 15-24 years in Perth). Eighty four per cent fell within National Heart Foundation "high risk" or "highest risk" categories for cardiovascular disease; 15% of men and 6% of women had an absolute risk of a cardiovascular event of over 15% within 10 years. A high proportion of participants reported diabetes, hypertension, smoking, overweight and obesity. A fasting plasma glucose level indicative of diabetes or impaired fasting glucose was found in 8.6% (95% CI, 6.2%-11%) of people not previously known to have diabetes. Obesity and smoking were twice as prevalent in study participants as in the general population. Less than a third of subjects with hypertension and diabetes had attained recommended target levels for blood pressure reduction or glycaemic control, and only a third of those at high risk and one in six of those at highest risk had attained recommended lipid-level targets. CONCLUSIONS: A cardiovascular risk assessment program with strong community support in an urban Aboriginal population can identify a significant number of people with high cardiovascular risk who are candidates for intensive risk-factor reduction strategies. PMID- 12885284 TI - Aspirin for cardiovascular disease prevention. AB - SECONDARY PREVENTION: Aspirin provides benefit in nearly all groups of patients with clinical manifestations of coronary heart disease. This includes patients with evolving acute myocardial infarction or after recovery from myocardial infarction, with unstable or stable angina, and those who undergo coronary artery bypass grafting or coronary angioplasty. Aspirin provides benefit in patients with peripheral arterial disease. This includes patients with acute or previous history of ischaemic stroke or transient ischaemic attack, those with lower limb arterial insufficiency, and those who undergo grafting or angioplasty of peripheral arterial vessels. PRIMARY PREVENTION: People without symptoms but at increased risk of a coronary heart disease event (> 1% annual risk) may reduce this risk by taking low-dose aspirin. However, the decision to take aspirin requires detailed consideration of individual cardiovascular risk and the potential benefit versus harm of treatment, particularly bleeding. Aspirin should only be used to prevent a cardiovascular event in association with an overall program of lifestyle measures including healthy eating, cessation of smoking, control of blood pressure and regular physical activity. ASPIRIN FOR PREVENTION: Prevention benefits of aspirin in heart disease can be achieved with doses as low as 75-150 mg daily. Unwanted effects of aspirin include stomach upsets, activation of peptic ulcers, an increased tendency to bruising, allergic reactions and increased risk of major gastrointestinal and other bleeding, including intracranial haemorrhage. In general, the risk of bleeding increases with increasing dose of aspirin and when it is used in combination with non steroidal anti-inflammatory drugs or oral anticoagulants. PMID- 12885285 TI - Management of healthcare workers after occupational exposure to hepatitis C virus. AB - The increasing rate of hepatitis C virus (HCV) infection in the community means that there is increased risk of occupational exposure for healthcare workers. In metropolitan hospitals in Victoria, we found that 80-150 healthcare workers have occupational exposures from HCV-infected patients annually. As there is a 1.8%-3% risk of transmission of HCV from a needlestick injury, two to five healthcare workers are likely to acquire HCV each year in Victoria. These needlestick injuries pose a personal, legal and professional risk to healthcare workers and their patients. Recent information shows that early antiviral treatment of acute HCV infection has high cure rates. Current local and international protocols for management of healthcare workers exposed to HCV do not address these issues. We propose a management protocol after needlestick injury that is stratified according to the likelihood of HCV acquisition and potential risk of staff-to patient transmission, and that is consistent with the current legal and clinical context of HCV infection in Australia. PMID- 12885286 TI - Current priorities for adolescent sexual and reproductive health in Australia. AB - The sexual health needs of teenagers differ from those of adults. Young sexually active teenagers are at high risk of Chlamydia trachomatis genital infection and its complications. Teenage pregnancy continues to be a problem in Australia. Current preventive strategies and clinical services in this domain of adolescent health in Australia are deficient. Australia can learn from the innovative and effective strategies developed in various countries for preventing high-risk sexual behaviours in teenagers. PMID- 12885287 TI - Occupational exposure to HIV: response to a system failure. PMID- 12885288 TI - Stem cell therapies: a tale of caution. AB - One of the most exciting possibilities in human therapeutics is that stem cells (embryonic or adult) may compensate for cell loss in disease, with functional recovery. This has received considerable publicity in the lay press. Much work remains to be done to turn stem cell therapy into a practical reality for major degenerative diseases, especially those affecting the nervous system. Medical scientists and journalists should work together in ensuring that the general public has a realistic understanding of the likely time frame in which benefits from stem cell therapies will be realised. PMID- 12885289 TI - The specialist neurologist and the "new genetics". AB - The "new genetics" will require specialist physicians to deal with an increasing number of genetic issues. Huntington disease (HD) is a rare single gene adult onset fatal neurodegenerative disorder. It provides a model to illustrate the role of the specialist physician in the new genetics. DNA testing options in HD include diagnostic DNA tests to confirm a provisional diagnosis, and predictive or presymptomatic DNA tests to determine whether disease will develop in an at risk individual. The specialist physician is well positioned to interact with the genetics services by providing in-depth knowledge of the clinical implications. This will become particularly relevant as the more complex multifactorial disorders (eg, Alzheimer disease) are understood at the DNA level. For optimal use of the new genetics, a team approach is essential to ensure that all areas of expertise are covered. PMID- 12885290 TI - Nocardia asteroides pneumonia with bacteraemia. PMID- 12885291 TI - Outbreak of influenza-like illness [corrected] related to air travel. PMID- 12885292 TI - Factors affecting female or male consultant stress in an Australian teaching hospital. PMID- 12885293 TI - A simple intervention to improve hospital antibiotic prescribing. PMID- 12885294 TI - Debriefing: care and sympathy are not enough. PMID- 12885295 TI - Sulphonation of dehydroepiandrosterone and neurosteroids: molecular cloning, expression, and functional characterization of a novel zebrafish SULT2 cytosolic sulphotransferase. AB - By searching the zebrafish EST (expressed-sequence tag) database, we have identified two partial cDNA clones encoding the 5' and 3' regions of a putative zebrafish sulphotransferase (ST). Using the reverse transcription-PCR technique, a full-length cDNA encoding this zebrafish ST was successfully cloned. Sequence analysis revealed that this novel zebrafish ST displays 44%, 43% and 40% amino acid identity with mouse SULT2B1, human SULT2B1b and human SULT2A1 ST respectively. This zebrafish ST therefore appears to belong to the SULT2 cytosolic ST gene family. Recombinant zebrafish ST, expressed using the pGEX-2TK prokaryotic expression system and purified from transformed Escherichia coli cells, migrated as a 34 kDa protein upon SDS/PAGE. Purified zebrafish ST displayed a strong sulphonating activity toward DHEA (dehydroepiandrosterone), with a optimum pH of 9.5. The enzyme also exhibited activities toward several neurosteroids with differential K(m) and V(max) values. A thermostability experiment revealed the enzyme to be relatively stable over a temperature range between 20 degrees C and 43 degrees C. Among ten different divalent metal cations tested, Fe2+ and Cd(2+ exhibited small, but significant, stimulatory effects, whereas Hg2+ and Cu2+ displayed considerably stronger inhibitory effects on the DHEA-sulphonating activity of the enzyme. These results constitute the first study on the molecular cloning, expression, and characterization of a zebrafish cytosolic SULT2 ST. PMID- 12885296 TI - Quantitative screening of advanced glycation endproducts in cellular and extracellular proteins by tandem mass spectrometry. AB - Glycation of proteins forms fructosamines and advanced glycation endproducts. Glycation adducts may be risk markers and risk factors of disease development. We measured the concentrations of the early glycation adduct fructosyl-lysine and 12 advanced glycation endproducts by liquid chromatography with tandem mass spectrometric detection. Underivatized analytes were detected free in physiological fluids and in enzymic hydrolysates of cellular and extracellular proteins. Hydroimidazolones were the most important glycation biomarkers quantitatively; monolysyl adducts (N(epsilon)-carboxymethyl-lysine and N(epsilon) 1-carboxyethyl-lysine) were found in moderate amounts, and bis(lysyl)imidazolium cross-links and pentosidine in lowest amounts. Quantitative screening showed high levels of advanced glycation endproducts in cellular protein and moderate levels in protein of blood plasma. Glycation adduct accumulation in tissues depended on the particular adduct and tissue type. Low levels of free advanced glycation endproducts were found in blood plasma and levels were 10-100-fold higher in urine. Advanced glycation endproduct residues were increased in blood plasma and at sites of vascular complications development in experimental diabetes; renal glomeruli, retina and peripheral nerve. In clinical uraemia, the concentrations of plasma protein advanced glycation endproduct residues increased 1-7-fold and free adduct concentrations increased up to 50-fold. Comprehensive screening of glycation adducts revealed the relative and quantitative importance of alpha oxoaldehyde-derived advanced glycation endproducts in physiological modification of proteins-particularly hydroimidazolones, the efficient renal clearance of free adducts, and the marked increases of glycation adducts in diabetes and uraemia particularly free advanced glycation endproducts in uraemia. Increased levels of these advanced glycation endproducts were associated with vascular complications in diabetes and uraemia. PMID- 12885297 TI - SH2-containing inositol 5-phosphatases 1 and 2 in blood platelets: their interactions and roles in the control of phosphatidylinositol 3,4,5-trisphosphate levels. AB - Src homology domain 2-containing inositol 5-phosphatases 1 and 2 (SHIP1 and SHIP2) are capable of dephosphorylating the second messenger PtdIns(3,4,5) P3 (phosphatidylinositol 3,4,5-trisphosphate) and interacting with several signalling proteins. SHIP1 is essentially expressed in haematopoietic cells, whereas SHIP2, a closely related enzyme, is ubiquitous. In the present study, we show that SHIP1 and SHIP2 are expressed as functional PtdIns(3,4,5) P3 5 phosphatases in human blood platelets and are capable of interacting when these two lipid phosphatases are co-expressed, either naturally (platelets and A20 B lymphoma cells) or artificially (COS-7 cells). Using COS-7 cells transfected with deletion mutants of SHIP2, we demonstrate that the Src homology domain 2 of SHIP2 is the minimal and sufficient protein motif responsible for the interaction between the two phosphatases. These results prompted us to investigate the relative importance of SHIP1 and SHIP2 in the control of PtdIns(3,4,5) P3 levels in platelets using homozygous or heterozygous SHIP1- or SHIP2-deficient mice. Our results strongly suggest that SHIP1, rather than SHIP2, plays a major role in controlling PtdIns(3,4,5) P3 levels in response to thrombin or collagen activation of mouse blood platelets. PMID- 12885298 TI - An automated method for high-throughput protein purification applied to a comparison of His-tag and GST-tag affinity chromatography. AB - BACKGROUND: Functional Genomics, the systematic characterisation of the functions of an organism's genes, includes the study of the gene products, the proteins. Such studies require methods to express and purify these proteins in a parallel, time and cost effective manner. RESULTS: We developed a method for parallel expression and purification of recombinant proteins with a hexahistidine tag (His tag) or glutathione S-transferase (GST)-tag from bacterial expression systems. Proteins are expressed in 96-well microplates and are purified by a fully automated procedure on a pipetting robot. Up to 90 microgram purified protein can be obtained from 1 ml microplate cultures. The procedure is readily reproducible and 96 proteins can be purified in approximately three hours. It avoids clearing of crude cellular lysates and the use of magnetic affinity beads and is therefore less expensive than comparable commercial systems. We have used this method to compare purification of a set of human proteins via His-tag or GST-tag. Proteins were expressed as fusions to an N-terminal tandem His- and GST-tag and were purified by metal chelating or glutathione affinity chromatography. The purity of the obtained protein samples was similar, yet His-tag purification resulted in higher yields for some proteins. CONCLUSION: A fully automated, robust and cost effective method was developed for the purification of proteins that can be used to quickly characterise expression clones in high throughput and to produce large numbers of proteins for functional studies.His-tag affinity purification was found to be more efficient than purification via GST-tag for some proteins. PMID- 12885299 TI - Pulmonary function testing in HTLV-I and HTLV-II infected humans: a cohort study. AB - BACKGROUND: HTLV-I infection has been linked to lung pathology and HTLV-II has been associated with an increased incidence of pneumonia and acute bronchitis. However it is unknown whether HTLV-I or -II infection alters pulmonary function. METHODS: We performed pulmonary function testing on HTLV-I, HTLV-II and HTLV seronegative subjects from the HTLV outcomes study (HOST), including vital capacity (VC), forced expiratory volume in one second (FEV1), and diffusing lung capacity for carbon monoxide (DLCO) corrected for hemoglobin and lung volume. Multivariable analysis adjusted for differences in age, gender, race/ethnicity, height and smoking history. RESULTS: Mean (standard deviation) pulmonary function values among the 257 subjects were as follows: FVC = 3.74 (0.89) L, FEV1 = 2.93 (0.67) L, DLCOcorr = 23.82 (5.89) ml/min/mmHg, alveolar ventilation (VA) = 5.25 (1.20) L and DLCOcorr/VA = 4.54 (0.87) ml/min/mmHg/L. There were no differences in FVC, FEV1 and DLCOcorr/VA by HTLV status. For DLCOcorr, HTLV-I and HTLV-II subjects had slightly lower values than seronegatives, but neither difference was statistically significant after adjustment for confounding. CONCLUSIONS: There was no difference in measured pulmonary function and diffusing capacity in generally healthy HTLV-I and HTLV-II subjects compared to seronegatives. These results suggest that previously described HTLV-associated abnormalities in bronchoalveolar cells and fluid may not affect pulmonary function. PMID- 12885300 TI - Longitudinal study on visual outcome and spectacle use after intracapsular cataract extraction in northern India. AB - PURPOSE: More than 3 million cataract extractions are undertaken in India annually. Almost 60% of these operations are intracapsular Cataract Extractions. The subsequent optical correction is provided by aphakic spectacles. The aim of this study is to assess visual outcome and perceived benefits of post-operative use of aphakic spectacles. METHODS: One hundred and sixty-seven persons who had undergone intracapsular cataract extraction and had been given best corrected aphakic spectacles were evaluated one year following prescription of the best corrected aphakic spectacles. Out of these, 82.6% were re-examined in this interview-based longitudinal study. RESULTS: The mean age of the male participants was 65.95 years and that of females was 71.26 years. 81.2% of the participants were using the provided spectacles. There was no significant association between the spectacle use and gender of the participant. The commonest reason stated by the respondents, for the non-use of the spectacles was 'poor vision'. 61.7% of the current users of provided spectacles had a visual acuity of 6/18 or better. 91.1% of the current users were very satisfied with the spectacles. All the current users could now manage personal activities and the spectacles had facilitated independent mobility. There was no difference in the level of satisfaction between mono-aphakics and bi-aphakics. Among the satisfied users, the modal spherical power was +10 D followed by +11 D. About one-third of these required a cylindrical correction. CONCLUSION: Following intracapsular cataract extraction, provision of the best correction after cataract surgery is desirable to obtain an optimal visual outcome. PMID- 12885304 TI - Contraceptive choices for women with inflammatory bowel disease. PMID- 12885301 TI - Congruence of tissue expression profiles from Gene Expression Atlas, SAGEmap and TissueInfo databases. AB - BACKGROUND: Extracting biological knowledge from large amounts of gene expression information deposited in public databases is a major challenge of the postgenomic era. Additional insights may be derived by data integration and cross-platform comparisons of expression profiles. However, database meta-analysis is complicated by differences in experimental technologies, data post-processing, database formats, and inconsistent gene and sample annotation. RESULTS: We have analysed expression profiles from three public databases: Gene Expression Atlas, SAGEmap and TissueInfo. These are repositories of oligonucleotide microarray, Serial Analysis of Gene Expression and Expressed Sequence Tag human gene expression data respectively. We devised a method, Preferential Expression Measure, to identify genes that are significantly over- or under-expressed in any given tissue. We examined intra- and inter-database consistency of Preferential Expression Measures. There was good correlation between replicate experiments of oligonucleotide microarray data, but there was less coherence in expression profiles as measured by Serial Analysis of Gene Expression and Expressed Sequence Tag counts. We investigated inter-database correlations for six tissue categories, for which data were present in the three databases. Significant positive correlations were found for brain, prostate and vascular endothelium but not for ovary, kidney, and pancreas. CONCLUSION: We show that data from Gene Expression Atlas, SAGEmap and TissueInfo can be integrated using the UniGene gene index, and that expression profiles correlate relatively well when large numbers of tags are available or when tissue cellular composition is simple. Finally, in the case of brain, we demonstrate that when PEM values show good correlation, predictions of tissue-specific expression based on integrated data are very accurate. PMID- 12885306 TI - Female sterilisation: is it what women really want? AB - OBJECTIVE: To audit the effectiveness of a female sterilisation counselling clinic. DESIGN: A prospective, observational study of a female sterilisation counselling clinic over a 6-month period to determine the proportions of attenders keeping their appointment, proceeding to sterilisation, and attending for surgery. SETTING: A hospital-based, city centre family planning clinic run by permanent, family planning-trained staff. RESULTS: A total of 226 women had appointments for female sterilisation counselling and most confirmed attendance. Only 153 (68%) women attended; of those 101 (66%) chose to be sterilised, the remainder choosing reversible contraception. Ninety-three (92%) women went ahead with the sterilisation procedure. Thus only 61% of the original attenders were sterilised; this was only 41% of those referred. CONCLUSIONS: The study suggests that many women referred for sterilisation counselling fail to attend. This finding needs to be further explored. For those women who do attend, accurate information and informed counselling is important. Many women are still unaware of highly effective long-term reversible contraception, and following discussion decide against female sterilisation. Services offering counselling for female sterilisation should also be able to provide easy access to a full range of contraceptive methods. PMID- 12885307 TI - Mifepristone and misoprostol for medical termination of pregnancy: the effectiveness of a flexible regimen. AB - BACKGROUND: Mifepristone, followed 48 hours later by administration of misoprostol, is a well-established regimen for medical termination of pregnancy (TOP). Although this regimen is effective, its inflexibility may limit its provision in an outpatient service. OBJECTIVE: To confirm that misoprostol administration is effective whether administered 24, 48 or 72 hours after oral mifepristone. DESIGN: Observational study of 234 consecutive women with pregnancies up to 83 days' gestational age in whom medical TOP was performed during the period December 2000-July 2001. SETTING: Women's Health Care Department, Royal Bolton Hospital, Bolton, UK. RESULTS: There was a high success rate for complete abortion in all groups whether mifepristone was administered 24, 48 or 72 hours prior to misoprostol. CONCLUSION: This study suggests that a more flexible regimen of mifepristone/misoprostol administration for medical TOP is effective in routine clinical practice. PMID- 12885308 TI - Providing information for young people in sexual health clinics: getting it right. AB - BACKGROUND: The need to improve the quality and availability of information on sexual health is identified as a key element in achieving the aims set out in the National Strategy for Sexual Health and HIV. Providing information about sexual health to young people poses particular challenges because of the sensitive nature of the issues and because of the difficulties that young people may face in sourcing information and asking questions of professionals. PMID- 12885309 TI - Capacity to consent to sexual relationships in adults with learning disabilities. AB - People with learning disabilities used to be seen as asexual or promiscuous and were discouraged from expressing their sexuality. However, with the growth of the rights movement, attention has turned to the dilemma of how to both empower and protect people with learning disabilities in relation to their sexuality. A recent research project showed that, on average, adults with learning disabilities knew much less about sex and understood much less about sexual abuse than non-disabled young people aged 16 years. Sex education made a significant difference to their knowledge and understanding however. The implications of the study for services for people with learning disabilities and for definitions of capacity to consent to sexual relationships are discussed. PMID- 12885310 TI - Use and effectiveness of oral analgesia when fitting an intrauterine device. AB - OBJECTIVE: To explore (a). if analgesia taken before an intrauterine device (IUD) fitting actually lessens the discomfort of the fitting and (b). whether there is a relationship between anxiety and choosing to take pain relief before an IUD fitting. POPULATION: Women having an IUD fitted in participating clinics in Leeds between 1 April and 31 July 2001. METHOD: Advice was given to women who were having an IUD fitted to take analgesia before they came to the clinic if they wished. The women completed a questionnaire at the time of fitting to record anticipated discomfort, actual discomfort and the use of analgesia before the fitting. A second questionnaire was sent to women 6 months later. RESPONSE: A total of 109 women completed the first questionnaire and 54 completed the second questionnaire. FINDINGS AND DISCUSSION: Women who were more anxious and who anticipated that the procedure would be painful, even if it was not as bad as expected at the time of the fitting, will recall that it was a painful experience. Those who chose to take painkillers before the procedure may identify this group of women. Higher anxiety levels may contribute to the perceived discomfort. PMID- 12885311 TI - Exacerbation of Crohn's disease after insertion of a levonorgestrel intrauterine system: a case report. AB - The case report describes a relapse of Crohn's disease in a woman soon after the fitting of a levonorgestrel intrauterine system (IUS). Whilst the published evidence on the effects of levonorgestrel IUS in patients with pre-existing inflammatory bowel disease is limited, reports to date suggest that caution should be exercised when considering fitting an IUS in such women, even if there appears to be much to gain from an expected reduction in menorrhagia. PMID- 12885312 TI - Perforation with the GyneFix intrauterine implant: is there a common factor? AB - This report describes an asymptomatic perforation with the GyneFix intrauterine contraceptive implant. A review of all other reports of this complication has been performed. Analysis of these reports suggests prolonged amenorrhoea secondary to continuous progestogen use as a possible common predisposing factor. PMID- 12885313 TI - The impalpable Implanon: a case report. AB - This is a case report of an Implanon contraceptive device that was impalpable after insertion and a discussion of the management of the impalpable Implanon. PMID- 12885314 TI - The 'intimate' examination: time for a name change? PMID- 12885315 TI - The members' enquiry service: frequently asked questions. PMID- 12885316 TI - Desogestrel-only pill (Cerazette). PMID- 12885317 TI - Evidence-based reproductive health: testing times for treatments. PMID- 12885326 TI - Size does matter and Homeopathic treatment of premenstrual symptoms. PMID- 12885327 TI - More on how to remove a Chinese IUD. PMID- 12885328 TI - Vasectomy techniques. PMID- 12885329 TI - Incidence of chromosomal mosaicism in human embryos at different developmental stages analyzed by fluorescence in situ hybridization. AB - Chromosomal mosaicism has been reported in in vitro-cultured embryos at early cleavage stages, as well as in morulae and blastocysts. We have assessed the incidence and pattern of mosaicism during in vitro development of human embryos from early-cleavage stages to morula and blastocyst. Fifty spare embryos were fixed for fluorescence in situ hybridization (FISH) analysis for chromosomes X, Y, 13, 18, and 21 on days 2 or 3 (4- to 10-cell stage) (n = 16), on day 4 (morula stage) (n = 14), on day 5 (pre-expanded blastocyst) (n = 5), and the expanded blastocyst stages (n = 15). Blocked embryos (no cleavage observed within the last 24 hr) were not included. A total of 2367 cells were analyzed. Four early cleavage stage embryos were found uniformly diploid; all of the others were mosaic for the chromosomes analyzed (mean diploid nuclei 48.3% +/- 28.7). All of the embryos at more advanced developmental stages, except one fully normal morula, had mosaic chromosome constitutions, with an increase in the percentage of diploid cells in morulae, pre-expanded, and expanded blastocysts, respectively (mean diploid nuclei 78.6% +/- 11.7, 66.0% +/- 20.8, 79.6% +/- 12.8), in comparison with earlier stages. Hypotheses about the origin of mosaicism and embryo regulation mechanisms will be discussed. PMID- 12885330 TI - Mutational analysis of the human dihydropyrimidine dehydrogenase gene by denaturing high-performance liquid chromatography. AB - Mutations in the DPYD gene, which encodes dihydropyrimidine dehydrogenase (DPD), the rate-limiting enzyme in the catabolism of pyrimidines, are responsible for an inborn error of metabolism associated with thymine-uraciluria and neurological symptoms. Because the antimetabolite 5-fluorouracil (5-FU) is metabolized by the same enzyme, deficient DPYD alleles may also constitute a risk factor for severe toxicity following treatment with this anticancer drug. The aim of this study was to develop a comprehensive and rapid method to detect sequence variations within the DPYD gene. Using polymerase chain reaction (PCR) amplification and denaturing high-performance liquid chromatography (DHPLC), we established a protocol that makes it possible to screen all 23 exons of the DPYD gene and their exon-intron boundaries for both known and unknown mutations under identical conditions. A novel one-step PCR mutagenesis procedure was developed to generate heterozygous mutant amplicons as positive controls to optimize DHPLC detection of any sequence variation. DHPLC analysis was shown to result in mutation-specific elution profiles and to be able to distinguish different base changes within the same exon or different heterozygous combinations of mutations within the same exon. By analyzing the DPYD gene in 16 affected individuals, a total of 47 base changes were detected, representing eight known mutations and three novel intronic base changes. Sequence analysis confirmed all base changes detected. This method will be useful in identifying patients at risk for toxicity prior to 5-FU treatment, as well as in the analysis of individual patients with thymine-uraciluria. PMID- 12885331 TI - Characterization of the molecular defect in the ATP7B gene in Wilson disease patients from Yugoslavia. AB - Wilson disease (WD) is an autosomal recessive disorder of copper metabolism resulting from the absence or dysfunction of a copper transporting P-type ATPase (ATP7B). Approximately 150 mutations of the ATP7B have been identified to date. In this paper, we report the results of molecular characterization and genotype phenotype analysis, which we have carried out on 35 patients from Yugoslavia affected by WD. Using single-strand conformational polymorphism (SSCP) followed by direct sequencing, we characterized the molecular defect in 80% of WD chromosomes and found 11 different mutations, three of which are novel. The most common mutations that accounted for the molecular defect in 71.3% of WD chromosomes were H1069Q (48.9%), 2304-2305insC (11.4%), R616Q (5.7%), and A1003T (5.7%). The results produced in this paper indicate that the best strategy for mutation detection in Yugoslavian patients with WD is an SSCP analysis of exons 14, 8, 5, and 13, where most of the defects (73.1%) lie, followed by mutation analysis of the remaining exons in ATP7B in patients in whom the mutation was not detected by the finitial screening. These data can be used to develop straightforward genetic testing in this population or in other countries composed of a genetically mixed population like the United States, where a significant number of immigrants came from Central and Eastern Europe. PMID- 12885332 TI - Sialic acid storage disease and related disorders. AB - This paper gives an overview of the two sialic acid storage disorders, Salla disease and infantile sialic acid storage disease, and the related disorders cystinosis, sialuria, sialidosis, and galactosialidosis. Sialic acid storage disease and cystinosis are models for a deficient lysosomal transport of monosaccharides and amino acids, respectively. Several gene mutations leading to the production of the faulty membrane proteins sialin and cystinosin have been identified in recent years. Knowledge of the underlying pathophysiology is a prerequisite for future research projects, which will focus on the expression of the disease genes in living systems and the physical characterization of these proteins by X-ray crystallography and nuclear magnetic resonance spectroscopy. PMID- 12885333 TI - Professional and personal attitudes about access and confidentiality in the genetic testing of children: a pilot study. AB - The ability to perform predictive genetic testing of children raises ethical concerns regarding whether and when to test and the disclosure of results. Semi structured interviews with a convenience sample of pediatricians (12) and geneticists (13) were conducted to see how they would react to parental requests for predictive genetic testing of their children, and their attitudes about testing their own children. We also asked about disclosure attitudes and practices for their patients' relatives and within their own families. Respondents would provide predictive genetic testing for most conditions, yet were less likely to seek this information about their own children. Respondents believed it was very important for patients to share some types of genetic information with relatives, and were directive in their counseling about intrafamilial disclosure, especially within their own families. Although respondents would almost never breach patient confidentiality, many would breach confidentiality within their own families. Health care professionals distinguish between their professional and personal roles with regard to issues of access and confidentiality in predictive testing of children. They are willing to provide greater access and more confidentiality for their patients than within their own families. PMID- 12885334 TI - Issues in newborn screening. AB - Newborn screening aims at the earliest possible recognition of disorders so that intervention with effective treatment can prevent the most serious consequences of the disorder. However, of several thousand known genetic disorders, therapy is presently available for only a small proportion of them. Newborn screening was first applied to phenylketonuria (PKU). Presently, newborn screening programs have been implemented in 26 countries for different diseases. However, potential problems in newborn screening programs that make implementation of newborn screening programs difficult include quality assessment, concerns about professional and technical competence, and ethical considerations. It has been shown that in the process from sampling the newborn to reporting of the screening results most errors are made in the pre- and post-analytical phases. It appears that much more could be done for the fetus if genetic screening and diagnostics could be accomplished early in pregnancy rather than after birth. PMID- 12885335 TI - A rapid and definitive test for Charcot-Marie-Tooth 1A and hereditary neuropathy with liability to pressure palsies using multiplexed real-time PCR. AB - Alterations in gene copy number have been shown to cause disease in humans. Two of the most common inherited peripheral neuropathies, Charcot-Marie-Tooth 1A (CMT1A) and hereditary neuropathy with liability to pressure palsies (HNPP), are two such diseases resulting from alteration in gene copy number of the dosage sensitive peripheral myelin protein 22 (PMP22) gene. Many complicated and laborious diagnostic tests exist for the diagnosis of these diseases. The aim of our study was to develop the first quantitative multiplex real-time PCR assay for the diagnosis of CMT1A and HNPP. A total of 160 individuals who were known to have CMT1A, HNPP, or were normal from previous testing were assayed by our multiplex real-time PCR method. The results confirmed the previously determined gene copy number of all patient and control individuals tested. The range of ratio values between the disease and control groups were easily defined. The assay is accurate, simple, and cost effective and can detect a 50% change in gene copy number. This represents an ideal assay for any small diagnostic laboratory. PMID- 12885336 TI - Screening for familial dysautonomia in Israel: evidence for higher carrier rate among Polish Ashkenazi Jews. AB - Familial dysautonomia (FD) is an autosomal recessive disorder characterized by hereditary sensory and autonomic neuropathies. Although extremely rare in most populations, FD is common among Ashkenazi Jews (AJ), with a calculated carrier frequency of 1 in 30, based on disease prevalence. The gene for FD was recently identified as IKBKAP. One major mutation (IVS2 + 6T --> C) is responsible in >99.5% of cases among AJ. The purpose of this study was to determine the actual frequency of FD carriers in the AJ population in Israel and to determine whether carriers are more frequent among a subpopulation of AJ from Poland. The study group included 1267 Jews of Ashkenazi origin who were referred for routine DNA screening tests. These included 1100 individuals who were full AJ and 167 who were part AJ. None had a family history of FD. Mutation analysis for (IVS2 + 6T - > C) was performed by PCR amplification followed by restriction enzyme analysis. All positive cases were confirmed by DHPLC WAVE( trade mark ). Among the 1100 full AJ tested, 34 were found to be FD carriers (1:32). The incidence of mutation carriers was significantly higher in AJ of Polish descent (1:18) compared to AJ of non-Polish descent (1:99). Among the 167 individuals who were part AJ, there were 3 carriers (1:56). The incidence of FD among AJ, particularly those of Polish background, warrants population screening. Population screening may be performed by denaturing high-performance liquid chromatography. PMID- 12885337 TI - High incidence of profound deafness in an isolated community. AB - In a Muslim Israeli Arab village, different types of hearing loss affect some 2% of the inhabitants. Most cases of profound deafness are due to recessive mutations in the Connexin-26 gene. Since in this community, marriages are by preference within the family (consanguineous), for many of the couples from the village the risk for an affected child is high. There are 30 families living in the village in which both parents have normal hearing and at least one child has a profound hearing defect. In these families, the birth of a child with profound deafness did not change family planning. The rate of marriage was similar for the siblings of deaf children as for other individuals in the village. The major problems were encountered by the deaf individuals themselves; in particular, most of the women were not married. Because of the distinctive nature of this particular problem, different types of screening programs were envisaged. However, all of them are problematic. Therefore, as a first step it was decided to begin a program including individual genetic counseling together with education of the entire population on practical aspects of human genetics. PMID- 12885338 TI - A simple method of screening for the common connexin-26 gene 35delG mutation in nonsyndromic neurosensory autosomal recessive deafness. AB - Mutations in the Connexin-26 gene are responsible for up to 60% of nonsyndromic, neurosensory autosomal recessive deafness (NSRD). Amongst all the mutations described to date, 35delG (a deletion of a G in a tract of five Gs at positions 30-35) is the most common and has been found in virtually all of the populations studied. Because its frequency varies in different populations, a rapid and simple method of detection of this mutation would be very helpful in population studies. A wide variety of methods for this detection have been described, but we herein present a very simple method using a PCR with primers designed to provide an amplicon of 94 or 93 nucleotides for the normal or mutant alleles, respectively, that can be easily distinguished in an 8% polyacrylamide gel. The entire protocol can be completed in a morning, thus supporting multiple runs. This assay will be useful in screening the large sample sizes required for population studies. PMID- 12885339 TI - Connexin-30 deletion analysis in connexin-26 heterozygotes. AB - Mutations in the Connexin-26 gene (Cx 26, GJB2) are the most common cause of hereditary nonsyndromic sensorineural hearing loss (SNHL). DNA analysis of the Cx 26 gene in deaf or hard-of-hearing individuals frequently demonstrates heterozygosity despite the fact that most mutations are known to be recessive. A 342-kb deletion in a gene adjacent to Cx 26, the Connexin-30 gene (Cx 30, GJB6), has been reported to cause deafness in the homozygous state or in combination with heterozygous mutations in Cx 26 (digenic inheritance). We have analyzed deaf or hard-of-hearing Cx 26 heterozygotes and individuals with no mutations in Cx 26 for this Cx 30 deletion. We found that 4/20 (20%) of the Cx 26 heterozygotes are heterozygous for this deletion and that no individuals were homozygous for the Cx 30 deletion. Cx 30 deletion analysis is recommended for all individuals with nonsyndromic SNHL following Cx 26 sequencing that does not demonstrate two recessive mutations. PMID- 12885340 TI - HFE alleles in an Irish cystic fibrosis population. AB - The variable clinical manifestations of cystic fibrosis (CF) suggest the influence of modifier genes. Genetic and environmental factors that determine whether an individual will develop associated complications are still being determined. It has been proposed that the gene for hemochromatosis, HFE, may be a modifier locus for CF disease phenotype. Recent research has suggested a relationship between mutations to the HFE gene and the development of meconium ileus (MI) and liver disease in CF. This study aims to expand our knowledge of the HFE mutations C282Y and H63D carrier rate in an Irish population of CF allele carriers. PCR restriction enzyme analysis was performed on blood samples from CF patients to identify the C282Y and H63D mutations. HFE status of CF allele carriers and CF patients (Delta F508) homozygotes with and without meconium ileus was determined. The carrier frequency for C282Y was 30.8% for the Delta F508 homozygote MI positive group, as compared to 12.5% for the non-Delta F508 MI positive group but did not reach statistical significance (p = 0.27). Interestingly, no Delta F508 homozygote patients were homozygous for the C282Y mutation. PMID- 12885341 TI - Absence of the Kilifi mutation in the rhinovirus-binding domain of ICAM-1 in a Caucasian population. AB - Human rhinoviruses (HRV), responsible for approximately 60% of the common colds, are divided into two groups, according to their receptor specificity. The major group of HRVs gains access to human cells by binding to the intercellular adhesion molecule-1 (ICAM-1), whereas HRVs of the minor group use members of the low-density lipoprotein receptor (LDLR) family for cell entry. Previous studies confirmed that the HRV-binding region of ICAM-1 is located in the amino-terminal immunoglobulin-like (Ig) domain 1, which is encoded by exon 2 of the ICAM-1 gene. An A --> T transversion in codon 29 of ICAM-1 exon 2 causes a lysine to methionine substitution (K29M), and was found at a high frequency (33.2%) in Kilifi (Kenya), as well as in other African populations. In this study we examined whether polymorphisms in exon 2 of ICAM-1 could be detected in a Caucasian population, assuming that these could be of importance in HRV binding. DNA from 100 healthy, unrelated, Belgian volunteers was obtained through a noninvasive swish-and-spit method. Using a primer set in the adjacent intron sequences, the full-length ICAM-1 exon 2 was amplified by polymerase chain reaction (PCR), followed by direct sequencing of the PCR product. No polymorphisms could be demonstrated in exon 2 of the ICAM-1 gene among all 100 tested individuals. The rhinovirus-binding Ig domain 1 of ICAM-1 seems to be a highly conserved region in the Caucasian population. PMID- 12885342 TI - Molecular genetic testing of beta-thalassemia patients of Indian origin and a novel 8-bp deletion mutation at codons 36/37/38/39. AB - Hemoglobinopathies are the most commonly inherited genetic disorders in India. Certain communities in India have a high predisposition to beta-thalassemia. To offer prenatal diagnosis and to prevent the birth of an affected child, mutation testing in clinically diagnosed beta-thalassemia patients/carriers is a prerequisite. Over a period of 4 years, we have conducted DNA analysis in 385 carriers for 15 beta-thalassemia mutations, HbD, HbE, and HbS. Using reverse dot blot (RDB) and amplification refractory mutation system (ARMS), we have been able to identify mutations in 381 of 385 thalassemia chromosomes. The study included the analysis of five common mutations found in Asian Indians, namely IVS1-5 (G C), 619-bp deletion, IVS1-1 (G-T), and the frameshifts at CD8/9(+G) and CD41/42( CTTT). The occurrence of these five mutations was seen in 299 (91.2%) carriers referred to us, the percentage of mutations varying between 4.0 and 68.9%. We also found Cd16 (-C) in 2.1%, CD30 (G-C) in 1.5%, and CD 15(G-A) in 0.6%; these are considered common mutations in the Indian population, as well. The beta thalassemia anomaly in 4 (0.6%) carriers remained uncharacterized by RDB and ARMS analysis. During delineation of the mutations in uncharacterized carriers by single-stranded conformational polymorphism (SSCP) and sequencing analysis, we have also been able to identify two unusual mutations, one involving an initiation codon and the second involving a novel 8-bp deletion, in Indian families of Uttar Pradesh origin. PMID- 12885343 TI - Chromosome 5. PMID- 12885344 TI - Safety of interleukin-12 gene therapy against cancer: a murine biodistribution and toxicity study. AB - As a prerequisite for a human clinical trial using interleukin (IL)-12 gene therapy, the biodistribution and safety of IL-12, administered as an intradermal naked DNA injection, was evaluated in mice. The pNGVL3-mIL12 plasmid used in this study is a nonviral vector designed to induce a high level of IL-12 protein expression during a transient transfection of the host cell. The biodistribution was evaluated by a polymerase chain reaction (PCR) assay that is capable of detecting less than 100 copies of the plasmid in the context of host DNA. Twenty four hours after three intradermal injections of 0.5 microg or 5 microg of pNGVL3 mIL12 plasmid, the plasmid was detectable in various internal organs, the blood, and the injection site. The plasmid was detectable in the gonads of only one animal at the high-dose treatment 24 hr after the injections. In the majority of the organs the plasmid was undetectable throughout the study. Possible side effects were monitored by histology and clinical chemistry, and the level of IL 12 protein expression was assessed by enzyme-linked immunosorbent assay (ELISA). No treatment-related histologic abnormalities were detected and the blood chemistry parameters showed no toxicity. The IL-12 protein was undetectable at all times at the injection site and interferon (IFN)-gamma levels at the injection site and in the serum were at background levels. The results of this murine safety study indicate that based on the distribution pattern of the plasmid in the body and the undetectable toxicities in the tissues, the use of the pNGVL3-hIL12 plasmid in cancer gene therapy clinical trials can be considered as safe. PMID- 12885345 TI - Improved packaging system for generation of high-level noncytotoxic HSV-1 amplicon vectors using Cre-loxP site-specific recombination to delete the packaging signals of defective helper genomes. AB - Amplicons are promising helper-dependent HSV-1-derived vectors that allow the transfer and expression of very large foreigner DNA into dividing and quiescent cells. We had already described an approach to prepare large amounts of high titer amplicon vectors, using Cre-loxP site-specific recombination system to delete the packaging ("a") signals of an HSV-1 recombinant helper virus (HSV-1 LaL). Amplicon vectors prepared using such a system showed a level of contamination with helper particles lower than 1%. The residual helper particles generated by this system are, however, replication-competent, thus precluding their use in gene therapy. To avoid such potential spread of residual particles, we present here the development of a defective Cre-loxP-based helper virus (HSV-1 LaL Delta J), deleted of the genes encoding ICP4 and ICP34.5 proteins from the helper genome, in addition to the native "a" signals. HSV-1 LaL Delta J carries a single floxed "a" signal in gC locus. To produce HSV-1 LaL Delta J and to prepare the amplicon vectors, we have constructed two novel cell lines expressing the essential ICP4 protein, either alone or in combination with Cre recombinase. These cell lines were conceived to complement ICP4 while minimizing the probability of generating replication-competent particles. In this paper we present results demonstrating that the novel helper system allows ready production of large amounts of high-titer amplicon vectors. Residual helper particles generated still do not exceed 0.5% of the viral population and can grow only in cells expressing ICP4. Amplicon vectors produced with this method showed no cytotoxicty for infected cells. PMID- 12885346 TI - Antiangiogenic cancer therapy with microencapsulated cells. AB - Inhibition of angiogenesis has led to tumor suppression in several cancer models. Although administering purified recombinant antiangiogenic product is effective, alternative approaches through genetic manipulation may be more cost-effective. We propose to implant nonautologous recombinant cells secreting angiostatin for systemic delivery of angiostatin in cancer treatment. These cells are protected from graft rejection in alginate microcapsules to function as "micro-organs" to deliver angiostatin in vivo. This approach was tested by implanting encapsulated mouse myoblast C2C12 cells genetically modified to secrete angiostatin into mice bearing solid tumor. Angiostatin was detected in sera of the treated mice. Efficacy was demonstrated by suppression of palpable tumor growth and improved survival. At autopsy, angiostatin localized to residual tumors and high levels of angiostatic activity were detected in tumor extracts. Tumor tissues showed increased apoptosis and necrosis compared with those from untreated or mock treated mice. Immunohistochemical staining against von Willebrand factor, an endothelial cell marker, showed that within tumors from the treated mice, the neovasculature was poorly defined by endothelial cells, many of which were undergoing apoptosis. However, the tumors eventually developed neovasculature independent of endothelial cells. Such vascular mimicry would account for the lack of long-term efficacy despite persistent angiostatin delivery. In conclusion, implantation with nonautologous microencapsulated cells is feasible for systemic delivery of angiostatin, resulting in localization of angiostatin to tumors and targeted apoptosis of the endothelial cells. Clinical efficacy was demonstrated by suppression of tumor growth and extension of life span. Although the potential of this cell-based approach for angiostatin-mediated cancer therapy is confirmed, long-term efficacy must take into account the possible escape by some tumors from angiogenesis inhibition. PMID- 12885347 TI - Phase I trial of intranasal and endobronchial administration of a recombinant adeno-associated virus serotype 2 (rAAV2)-CFTR vector in adult cystic fibrosis patients: a two-part clinical study. AB - Recombinant adeno-associated serotype 2-based vectors (rAAV2) possess a number of theoretical advantages for cystic fibrosis (CF) gene therapy because they elicit little or no inflammatory response and generally result in stable expression. rAAV2 vectors expressing the cystic fibrosis transmembrane conductance regulator (CFTR) gene have previously been shown to mediate stable correction of the CF defect in CF bronchial epithelial cells and stable expression of CFTR in rabbit and nonhuman primate models. Here we report the results of the first trial initiated with rAAV in humans, a phase I study in 25 adult and adolescent CF patients with mild to moderate lung disease. Doses of the rAAV-CFTR vector (tgAAVCF) ranging from 3 x 10(1) to 1 x 10(9) replication units (RU), which is equivalent to approximately 6 x 10(4) to 2 x 10(12) DNase resistant particles (DRP), were administered to one side of the nose and to the superior segment of the lower lobe of the right lung. Several adverse events were noted prior to and/or after vector delivery, but most of them appeared to be related to the endogenous CF lung disease or a result of the bronchoscopic procedures. Only one of the serious events was judged to be possibly vector-related (based on temporal association), and this event was a pulmonary exacerbation very similar to several others experienced by the same subject in the three months preceding vector delivery. Vector shedding was minimal throughout the study, and serum neutralizing antibodies were detected after vector delivery to subjects in the highest dosage cohorts. Gene transfer as measured by DNA polymerase chain reaction (PCR) was not observed until cohort 10 in nasal and bronchial epithelia. Sporadic low-level copy numbers suggested gene transfer of anywhere from 0.002 copies per cell up to 0.5 copies per cell was possible; however, DNA PCR was positive in lungs prior to direct dosing suggesting aspiration from the nasal dosing. These data indicate the need for continued evaluation of rAAV-CFTR vectors in additional clinical trials. PMID- 12885348 TI - Lentivirus-mediated gene transfer and expression in established human tumor antigen-specific cytotoxic T cells and primary unstimulated T cells. AB - In this report, we evaluated the efficiency of stable gene transfer into established CD8(+) human tumor antigen-specific cytotoxic T cell (CTL) lines and peripheral blood lymphocytes (PBL) by oncoretroviral and lentiviral vectors. In the oncoretroviral vector, the green fluorescent protein (GFP) reporter gene was regulated by the murine stem cell virus (MSCV) promoter. In three human immunodeficiency virus type 1 (HIV-1)-based lentiviral vectors, the GFP transgene was regulated by either a chimeric MSCV/HIV-1 promoter, or cellular promoters from human housekeeping genes PGK and EF1 alpha. We found that several lines of proliferating tumor-specific CTL were poorly (=2%) transduced by the oncoretroviral vector that transduced Jurkat T cell line efficiently (=80%). In contrast, three lentiviral vectors transduced 38-63% of these proliferating CTL. More interestingly, all lentiviral vectors packaged without the HIV-1 accessory proteins transduced human bulk PBL and purified CD4(+) and CD8(+) lymphocyte subsets without prior stimulation. Detailed analysis indicated that the lentiviral vectors containing the EF1 alpha or PGK ubiquitous promoter can transduce unstimulated PBL and achieve low-level transgene expression in the absence of any T-cell activation. However, T-cell activation subsequent to the transduction of unstimulated PBL is required for high-level transgene expression. Transduced PBL expressing transgene delivered by the lentiviral vectors still preserved resting and naive cell phenotypes. Taken together, prior T cell stimulation and HIV-1 accessory proteins are dispensable for lentivirus-mediated gene transfer into resting naive and memory T lymphocytes. These results will have significant implications for the study of T-cell biology and for the improvement of clinical gene therapies of acquired immune deficiency syndrome (AIDS) and cancer. PMID- 12885349 TI - Ex vivo gene therapy for skeletal regeneration in cranial defects compromised by postoperative radiotherapy. AB - Because radiation remains a common postoperative treatment for head and neck cancers, it is critical to determine whether new bone-regenerative approaches are effective for healing craniofacial defects challenged by therapeutic doses of radiation. The objective of this study was to determine whether the deleterious effects of radiotherapy could be overcome by ex vivo gene therapy to heal craniofacial defects. Rat calvarial critical-sized defects were treated with either an inlay calvarial bone graft or syngeneic dermal fibroblasts transduced ex vivo with an adenovirus engineered to express bone morphogenetic protein 7 (BMP-7), a morphogen known to stimulate bone formation. Two weeks postoperatively, either no radiation or a single 12-Gy radiation dose was delivered to the operated area and the tissue was harvested 4 weeks later. None of the inlay bone grafts healed at the wound margins of either the radiated or nonradiated sites. In contrast, bone was successfully regenerated when using an ex vivo gene therapy approach. More bone formed in the nonradiated group as determined by the percentage of defect surface covered (87 +/- 4.1 versus 65 +/- 4.7%; p = 0.003) and percentage of defect area filled by new bone (60 +/- 5.9 versus 32 +/- 2.7%; p = 0.002). Although the effects of radiation on the wound were not completely overcome by the gene therapy approach, bone regeneration was still successful despite the radiation sensitivity of the fibroblasts. These results indicate that BMP-7 ex vivo gene therapy is capable of successfully regenerating bone in rat calvarial defects even after a therapeutic dose of radiation. This approach may represent a new strategy for regenerating skeletal elements lost due to head and neck cancer. PMID- 12885350 TI - Vaccination of women with metastatic breast cancer, using a costimulatory gene (CD80)-modified, HLA-A2-matched, allogeneic, breast cancer cell line: clinical and immunological results. AB - MDA-MB-231, an HLA-A2(+), HER2/neu(+) allogeneic breast cancer cell line genetically modified to express the costimulatory molecule CD80 (B7-1), was used to vaccinate 30 women with previously treated stage IV breast cancer. Expression of CD80 conferred the ability to deliver a costimulatory signal and thereby improved the antigen presentation capability of the tumor cells to patient T cells in vitro. Patients were vaccinated with 10(7) or 10(8) irradiated gene modified tumor cells with granulocyte-macrophage colony-stimulating factor (GM CSF) or BCG, three times at 2-week intervals and then monthly until progressive disease developed. GM-CSF-related flulike symptoms and minor injection site reactions were observed frequently. Prolonged disease stabilization was observed in four patients but no objective tumor regressions were seen. Immune responses were measured in matched peripheral blood samples collected before and after treatment from 9 of 15 patients treated at the 10(8) tumor cell dose. Four patients exhibited MHC class I-restricted cytokine production in response to the parental breast cancer cell line. One patient maintained an increased number of circulating tumor-specific, interferon gamma-secreting CD8(+) T cells for 24 months after the last vaccination. One patient exhibited a tumor-specific interleukin 5 response to an autologous tumor cell line. This immunization strategy proved to be safe and feasible, and induced tumor-specific immune responses in a minority of patients; however, no objective tumor regressions were observed. PMID- 12885351 TI - Oxalate-degrading enzymes from Oxalobacter formigenes: a novel device coating to reduce urinary tract biomaterial-related encrustation. AB - BACKGROUND AND PURPOSE: The long-term placement of biomaterials within the urinary tract is limited by the development of encrustation. In a noninfected urinary environment, encrustation often results from the deposition of calcium oxalate on the biomaterial surface. There is an association between the absence of Oxalobacter formigenes, a commensal colonic bacterium capable of degrading oxalate, and calcium oxalate stone formation. This pilot study was designed to evaluate several oxalate-degrading enzymes produced by O. formigenes as a potential biomaterial coating to reduce urinary tract encrustation. MATERIALS AND METHODS: Circular silicone disks of 6-mm diameter were incubated for 48 hours in oxalylcoenzyme A decarboxylase (OXC), formyl-coenzyme A transferase (FRC), and coenzyme A, while control disks were incubated in distilled water. The adsorption of OXC and FRC was assessed using enhanced chemiluminescence (ECL) and atomic force microscopy (AFM). Coated and uncoated disks (20 of each) were implanted in the bladders of 40 female New Zealand White rabbits. After 30 days, the disks were recovered, and the degree of encrustation on the polymer surface was evaluated utilizing dry weight measurement, calcium atomic absorption spectroscopy (AAS), and scanning electron microscopy/energy-dispersive X-ray analysis (SEM/EDX). RESULTS: Both ECL and AFM demonstrated coating of the silicone disks with OXC and FRC. The mean dry weights of the coated and control disks following explantation were 0.591 +/- 0.438 g and 0.747 +/- 0.428 g, respectively (P = 0.307). The mean weight of calcium on the coated and control disks, as determined by AAS, was 154.1 +/- 96.25 mg and 258 +/- 181.35 mg, respectively (P = 0.008). CONCLUSIONS: The use of oxalate-degrading enzymes from O. formigenes to coat urinary biomaterials represents a novel paradigm to reduce biomaterial-related encrustation. Coating of silicone with oxalate-degrading enzymes from O. formigenes results in a modest reduction in encrustation with no apparent toxicity. Further studies are warranted. PMID- 12885352 TI - Laparoscopic inferior vena cava and right atrial thrombectomy utilizing deep hypothermic circulatory arrest. AB - BACKGROUND AND PURPOSE: Surgery for renal cancer associated with a level III or IV tumor thrombus often involves cardiopulmonary bypass, deep hypothermia, and exploration of the right atrium and inferior vena cava (IVC). This major open operation necessitates a large median sternotomy incision and a midline abdominal or chevron incision. Herein, we investigate the feasibility of purely laparoscopic IVC and right atrial thrombectomy utilizing deep hypothermic circulatory arrest. MATERIALS AND METHODS: In six male calves weighing 70 to 80 kg, the right common carotid artery and right internal jugular vein were cannulated for subsequent cardiopulmonary bypass. One laparoscopic team performed right radical nephrectomy and complete mobilization of the intra-abdominal IVC by a four-port approach. Simultaneously, a second laparoscopic team obtained three port thoracoscopic access to incise the pericardium and expose the right atrium. In sequence, cardiopulmonary bypass, complete exsanguination, cardiac arrest, and core hypothermia of 18 degrees C were achieved. A coagulum thrombus was created by needle injection into the IVC. Combined laparoscopic and thoracoscopic incision, exploration, and thrombectomy of the IVC and the right atrium were then performed in a bloodless field. An angioscope was inserted inside the heart and the IVC to confirm complete thrombus clearance visually. The IVC and right atrium were then laparoscopically suture repaired, cardiopulmonary bypass was reestablished, and the animal was gradually rewarmed. Once sinus rhythm was reestablished at normal body temperature, the animal was weaned off the pump. RESULTS: The mean total operative time was 494.5 minutes (range 355-705 minutes). The mean time needed to lower the core temperature was 63.5 minutes (range 50-120 minutes), and the mean time required to rewarm the animal was 101.8 minutes (range 70-130 minutes). The mean blood volume drained into the pump was 2633.3 mL (range 1400-3200 mL), and the mean estimated blood loss was 350 mL (range 200-750 mL). Reestablishment of sinus cardiac rhythm and weaning off the pump was successful in all animals prior to acute euthanasia. CONCLUSIONS: Laparoscopic radical nephrectomy with thrombectomy for level III or IV tumor thrombi utilizing deep hypothermic circulatory arrest is feasible in the calf model using minimally invasive techniques exclusively. The procedure is technically complex and requires the combined efforts of expert urologic and cardiac operative teams. Survival studies are planned. PMID- 12885353 TI - Ureteral tissue balloon expansion for laparoscopic bladder augmentation: survival study. AB - BACKGROUND AND PURPOSE: The search for the perfect urinary bladder substitute continues. Despite their inherent limitations, intestinal segments remain the commonest material for bladder reconstruction. The ureter, with its transitional epithelium, may be the ideal tissue to augment the bladder. Ikeguchi et al reported the feasibility of chronic ureteral balloon expansion by open surgery (J Urol 1998;159:1665). Herein, we propose a completely minimally invasive approach to balloon overdilate a segment of juxtavesical ureter incrementally and to use this in-line tissue-expanded ureteral patch to augment the bladder laparoscopically. MATERIALS AND METHODS: In five female pigs, a novel ureteral expansion balloon device (Microvasive, MA) was inserted percutaneously and advanced antegrade into the juxtavesical ureter. The device has two channels: one for balloon inflation and the other for draining the kidney. After progressive ureteral expansion over a 3- to 4-week period, laparoscopic augmentation ureterocystoplasty was performed. Animals were euthanized at 15 days (N = 1), 1 month (N = 1), 2 months (N = 1), and 3 months (N = 2). RESULTS: Percutaneous balloon device placement was technically successful in all five cases (mean operating room time 52 minutes). The mean volume of the tissue-expanded ureter at 1, 2, and 3 weeks was 12.9 cc, 60.3 cc, and 171.8 cc, respectively. Laparoscopic augmentation ureterocystoplasty with (N = 3) or without (N = 2) concomitant subtotal cystectomy was technically successful in all five cases without any open conversion. The mean operative time was 126.5 minutes, and the mean blood loss was 29 mL. Postoperative complications consisted of one case each of pyelonephritis and ureteral stricture. At autopsy, the mean capacity of the bladder was 574 mL, and the P(ves) at maximum capacity was 14 cm H(2)O. Histologic examination of the tissue-expanded ureter revealed regenerated transitional epithelium and muscle hypertrophy. CONCLUSIONS: Chronic ureteral tissue expansion can be carried out safely and efficaciously. The expanded tissue is thick, healthy, and vascular, with histologic features of normal transitional epithelium and muscle hypertrophy and hyperplasia. This expanded ureteral tissue can be used to augment the bladder with laparoscopic techniques. Such augmented bladders do not show significant shrinkage and possess urodynamic characteristic of normal capacity and normal compliance over a follow-up of 3 months. PMID- 12885354 TI - Sutureless laparoscopic heminephrectomy using laser tissue soldering. AB - BACKGROUND AND PURPOSE: Widespread application of laparoscopic partial nephrectomy has been limited by the lack of a reliable means of attaining hemostasis. We describe laser tissue welding using human albumin as a solder to control bleeding and seal the collecting system during laparoscopic heminephrectomy in a porcine model. MATERIALS AND METHODS: Laparoscopic left lower-pole heminephrectomy was performed in five female domestic pigs after occluding the hilar vessels. Using an 810-nm pulsed diode laser (20 W), a 50% liquid albumin-indocyanine green solder was welded to the cut edge of the renal parenchyma to seal the collecting system and achieve hemostasis. Two weeks later, an identical procedure was performed on the right kidney, after which, the animals were sacrificed and both kidneys were harvested for ex vivo retrograde pyelograms and histopathologic analysis. RESULTS: All 10 heminephrectomies were performed without complication. The mean operative time was 82 minutes, with an average blood loss of 43.5 mL per procedure. The mean warm ischemia time was 11.7 minutes. For each heminephrectomy, a mean of 4.2 mL of solder was welded to the cut parenchymal surface. In three of the five acute kidneys and all five 2-week kidneys, ex vivo retrograde pyelograms demonstrated no extravasation. In addition, no animal had clinical evidence of urinoma or delayed hemorrhage. Histopathologic analysis showed preservation of the renal parenchyma immediately beneath the solder. DISCUSSION: Laser tissue welding provided reliable hemostasis and closure of the collecting system while protecting the underlying parenchyma from the deleterious effect of the laser during porcine laparoscopic heminephrectomy. PMID- 12885355 TI - Laparoscopic ileocytoplasty and continent ileovesicostomy in a porcine model. AB - BACKGROUND AND PURPOSE: Bladder augmentation with autologous bowel is commonly accompanied by a continent ileovesicostomy (the Monti procedure). Herein, we describe our initial experience with laparoscopy-assisted ileocystoplasty and continent ileovesicostomy in a porcine model. MATERIALS AND METHODS: Thirteen Yucatan minipigs with an average weight 49 lb underwent a laparoscopy-assisted ileocystoplasty and ileovesicostomy (Monti procedure) using a four-port technique. After catheter removal at 3 weeks, the animals underwent twice-daily catheterization of the ileovesicostomy until complete occlusion of the ileovesicostomy occurred. RESULTS: All animals survived the initial surgery, with 10 animals being available for catheterization at 3 weeks. The average Monti anastomotic time, average ileal patch-bladder anastomotic time, and average total surgery times were 47 minutes, 89 minutes, and 307 minutes, respectively. Eight of ten animals developed stomal stenosis after catheter removal (average time to complete occlusion of 6 days; range 1-13 days), with two animals also demonstrating partial obstruction at the Monti-bladder anastomosis and four partial obstruction at the fascial level of the Monti tube. Pathologic review of select bladder augmentation and Monti tube specimens revealed moderate to severe acute and chronic inflammation with severe scarring at the Monti-skin stoma site in the majority of specimens. CONCLUSIONS: We have demonstrated the feasibility of laparoscopic ileocystoplasty and continent ileovesicostomy in a porcine model. The development of stomal stenosis may be related to the intrinsic properties of the pig skin, but additional investigation is required prior to human studies of this novel surgical technique. PMID- 12885356 TI - Simplified laparoscopic radical cystectomy with orthotopic ileal neobladder creation in a porcine model. AB - BACKGROUND AND PURPOSE: Laparoscopic radical cystectomy with orthotopic ileal neobladder creation is a technically challenging and lengthy surgical procedure. We present our experience with a simplified technique for laparoscopic cystectomy and neobladder creation in the porcine model. MATERIALS AND METHODS: Ten female minipigs underwent a purely laparoscopic radical cystectomy with orthotopic ileal neobladder creation. Nine ureterointestinal anastomoses were performed using a simplified "dunk" technique, where the ureter was prolapsed 5 mm into the afferent limb and the periureteral tissue was secured to the bowel serosa with three superficial sutures. Six ureters were not stented, and three had indwelling stents inserted. In 11 ureters, the anastomosis was performed using a running mucosa-to-mucosa technique (three with stents, eight without stents). The Lapra Ty suture clip (Ethicon Endosurgery, Cincinnati, OH) was used to secure the running sutures on the urethra, ureters, and neobladder. Animals were harvested at 3 to 8 weeks (mean 6.5 weeks) after surgery. Serology, static cystogram, intravenous urography, and gross and histopathologic evaluations were performed. RESULTS: Of six unstented dunked ureterointestinal anastomoses, two (33%) were widely patent, two were strictured but patent, and two were completely obstructed. In the three stented ureters implanted using the dunk technique, one (33%) was widely patent, one was strictured, and one was completely obstructed. All ureterointestinal anastomoses performed with a mucosa-to-mucosa running anastomosis, whether stented (three ureters) or not stented (eight ureters), were widely patent. Lapra-Ty clip migration into the neobladder pouch caused urethral obstruction resulting in delayed bladder perforation in two animals. CONCLUSIONS: Laparoscopic cystectomy and ileal neobladder creation is technically feasible. Attempts to simplify the ureterointestinal anastomosis require further evaluation and modification. Stent placement appears to be unnecessary in the laparoscopic ureterointestinal anastomosis. Laparoscopic creation of the ileal neobladder remains a technically challenging procedure. PMID- 12885357 TI - Comparison of laparoscopy-assisted partial nephrectomy using digital compression versus purely laparoscopic nephrectomy using vascular pedicle clamp in a porcine model. AB - BACKGROUND AND PURPOSE: Laparoscopic partial nephrectomy raises a number of technical issues, particularly in the control of bleeding. We compared digital compression during hand assistance with a vascular clamp. MATERIALS AND METHODS: Laparoscopic transperitoneal bilateral lower-pole partial nephrectomy was performed on six pigs, with a vascular clamp being used on one side and digital pressure through an Omniport or Lapdisk on the other. The pigs were then euthanized and the kidneys harvested for histologic examination. RESULTS: Good hemostasis was obtained regardless of the technique. However, the estimated blood loss was significantly greater with digital compression (96.7 versus 9.6 mL). There were no significant micropathological differences in the kidneys. The surgeons rated suture tying and placement much easier with the purely laparoscopic technique. CONCLUSIONS: Both techniques are feasible. In experienced hands, the purely laparoscopic technique takes no longer than the hand-assisted technique and is associated with less blood loss, easier suturing, and better cosmesis. However, long-term follow-up is needed before clinical use can be contemplated. PMID- 12885358 TI - Immune response to laparoscopic bowel injury. AB - BACKGROUND AND PURPOSE: Laparoscopic bowel injuries are rare but potentially fatal if recognition is delayed. Unlike the situation after open surgery, patients with unrecognized bowel injury after laparoscopy do not present with the typical "acute surgical abdomen." We investigated monocyte, neutrophil, and lymphocyte apoptosis as indicators of the immune response and whether this response is stimulated or suppressed by laparoscopic bowel injury compared with bowel injury induced during open surgery. MATERIALS AND METHODS: After an animal protocol was approved, laparoscopy was performed in a rabbit model. A total of 44 animals were divided into four groups of 11 rabbits each. Laparoscopic bowel injury was created using 30-W electrocautery at 0 (control), 1, and 5 hours after induction of pneumoperitoneum. Bowel injury was created in the fourth group during open laparotomy. Animals were euthanized at 0, 1 day, 1 week, or 2 weeks after surgery. Apoptosis was assessed by staining the nuclei of blood cells with H-33342 dye. RESULTS: At 1 week, neutrophil, monocyte, and lymphocyte apoptosis levels were 2.4- to 5-fold lower after laparoscopy (1-hour pneumoperitoneum) compared with open surgery. However, at 2 weeks, the percentage of apoptosis had equalized in the two groups. Interestingly, with longer laparoscopic procedures (5 hours), the percentage of apoptosis at 0 and 1 day more closely approached that seen after open surgery. At 2 weeks, there was a significant difference in apoptosis levels in all cell types between the experimental groups compared with controls (P < 0.001). No animals undergoing a 5-hour open procedure survived to 2 weeks after bowel injury. CONCLUSIONS: Open surgery resulted in a significant increase in programmed cell death compared with controls in the immediate postoperative period following bowel injury. Laparoscopic surgery produced a delayed response and after 2 weeks with bowel perforation approached open surgery levels. The difference in the degree of cellular death may be secondary to a smaller degree of stimulation of the immune response in laparoscopic surgery. PMID- 12885359 TI - In vitro training program to improve ambidextrous skill and reduce physical fatigue during laparoscopic surgery: preliminary experience. AB - PURPOSE: We illustrate the preliminary experience with our planned intensive in vitro training program focused on the nondominant hand and reducing physical fatigue. Apart from this, we tried to calculate how much training is required for a novice urologist to master laparoscopic freehand suturing with the dominant and nondominant hands. MATERIALS AND METHODS: Between December 2001 and May 2002, one trainee worked on a Pelvi-trainer, first with the dominant (right) hand and then with the left hand, practicing intracorporeal suturing and cutting and improving physical endurance by gradually increasing the duration of each training session. Along with the Pelvi-trainer, he worked on left-handed writing to improve wrist movements. Before starting this training, he did not have any laparoscopic experience as assistant or primary surgeon. The progress was stored in the computer prospectively to compare the results with those obtained at the end of the training. RESULTS: For the dominant hand, 30 hours over 2 months was required to master laparoscopic suturing skills. After training of the right hand, the nondominant hand required 40 hours of Pelvi-trainer work and 20 hours of handwriting during 2 months. Physical endurance for suturing in the Pelvi-trainer increased from 15 minutes to 150 minutes over 3 months. The main improvement was in the degree of pain over the right shoulder (because of the abducted position) and backache. CONCLUSION: Incorporation of sufficient in vitro training to improve nondominant hand functioning and reduce physical fatigue can make adaptation to laparoscopic surgery easier. PMID- 12885360 TI - Image converter eliminates mirror imaging during laparoscopy. AB - BACKGROUND AND PURPOSE: In laparoscopy, the term "mirror imaging" is used to describe a visual illusion resulting in paradoxical movements when a surgeon is positioned opposite the laparoscope. Mirror imaging is a common problem, creating difficulty in ergonomics and task performance. We introduce the use of a video image converter box (IC box) to overcome mirror imaging. The IC box converts the analog signal to a digital one, performs image rotation or inversion or both, and then reproduces an analog signal for monitor viewing. A laboratory study evaluated whether the IC box could improve performance during laparoscopic tasks. MATERIALS AND METHODS: Fourteen laparoscopic surgeons (10 novice and 4 experienced) completed three laparoscopic tasks while positioned opposite the camera and experiencing mirror imaging: (1). suture cutting; (2). multiple transfers of a piece of foam; and (3). multiple transfers of a pinto bean. Participants were timed during each test both with and without the use of the IC box. RESULTS: All surgeons completed each task faster using the IC box (P < 0.015). On average, use of the IC box allowed subjects to complete assignments in less than one-third the time needed without the IC box. In Task 3, requiring multiple transfers of a small bean, all participants using the IC box completed the task. However, without the box, only 1 of 14 participants accomplished the goal in the allotted time. CONCLUSION: Use of the IC box eliminates mirror imaging and improves performance and efficiency during laparoscopic tasks. The box would significantly benefit surgeons positioned opposite the camera during laparoscopic surgery. PMID- 12885361 TI - In vivo proteomic analysis of cytokine expression in laser capture-microdissected urothelial cells of obstructed ureteropelvic junction procured by laparoscopic dismembered pyeloplasty. AB - BACKGROUND AND PURPOSE: Ureteropelvic junction obstruction (UPJO) is defined as an impediment to urinary flow from the renal pelvis into the ureter. The exact cause remains an enigma despite investigations along embryologic, anatomic, and histologic lines. Our goal was to investigate in vivo the expression profile of cytokines in hyperplastic urothelial cells as a means of determining the source of UPJO. MATERIALS AND METHODS: Cellular proteomes of matched normal and hyperplastic urothelial cells were analyzed by laser capture microdissection (LCM) and tissue microdissection and human cytokine proteomic chips. All specimens (N = 9) were surgically obtained from patients undergoing laparoscopic dismembered pyeloplasty and were immediately embedded in O.C.T. solution and flash-frozen in liquid nitrogen. Tissue sections (6 microm) were mounted on uncoated glass slides using a cryostat, fixed in 70% ethanol, stained with hematoxylin and eosin, and sequentially dehydrated in ethanol and xylene. Typically, paired samples of normal and hyperplastic urothelial cells were procured on separate caps from serial sections of each specimen by the Arcturus PixCell II LCM system using 3000 laser pulses and a spot diameter of 30 microm. Total proteins were harvested and quantitated. Differential expression profile analysis of 43 cytokines in normal and hyperplastic cells were performed using human protein chips. Briefly, the membranes were initially probed with protein (150 ng) from normal or hyperplastic cells and sequentially reacted with a cocktail of biotinylated cytokine antibodies and horseradish peroxidase conjugated streptavidin. The membranes were developed using enhanced chemiluminescence and analyzed by densitometry. RESULTS: Comparative densitometric analysis revealed twofold to fourfold upregulation of growth related oncogene alpha (GRO-alpha), interleukin (IL)-1alpha, interferon (INF) gamma, IL-8, tumor necrosis factor (TNF)-alpha, RANTES, and macrophage inflammatory protein-1beta (MIP-1beta) and twofold to fourfold downregulation of transforming growth factor (TGF)-beta and IL-10 in hyperplastic urothelial cells compared with paired control cells. CONCLUSIONS: We here report the efficient application of LCM and proteomic array chips for expression profile analysis of cytokines in vivo. Because the etiology and pathogenesis of UPJO are still fragmentary, the marked heterogeneity of the observed cytokine alterations reported here may be of significance. Further studies are required to elucidate the functional significance of the differentially expressed cytokines in the pathogenesis of the disease. PMID- 12885362 TI - [Tegument viral protein 22 enhanced cell-killing effect of the herpes simplex virus thymidine kinase/ganciclovir system on ovarian cancer in vivo]. AB - OBJECTIVE: To explore the enhanced cell-killing effect in vivo of herpes simplex virus thymidine kinase/ganciclovir (HSV-TK/GCV) gene therapeutic system using tegument viral protein (VP22) intercellular trafficking. METHODS: Human ovarian epithelial cancer cell line 3AO was infected by lentivirus containing HSV-TK and HSV-VP22-TK respectively. Tumors were induced in nude mice by subcutaneous injection of the mixture of 90% 3AO cells and 10% 3AO cells carried with HSV-TK (3AO/TK) or HSV-VP22-TK (3AO/VP22-TK). Nude mice injected with 3AO cells were used as blank control. When the volume of tumor was 150 mm(3), GCV was administered at 10 mg/kg or 50 mg/kg intraperitoneally. RESULTS: There were significant differences, in the tumor volume and weight between 3AO/TK group and 3AO/VP22-TK group after administration of 10 mg/kg GCV (P < 0.01), and the later was more efficient than the former. But there was no significant difference after administration of 50 mg/kg GCV (P > 0.05). The tumor inhibition rates in 3AO/VP22 group and 3AO/VP22-TK group were 37.7% and 91.5% respectively after administration of 10 mg/kg GCV (P < 0.01), and were 81.8% and 96.7% respectively after administration of 50 mg/kg GCV (P > 0.05). CONCLUSION: These results clearly indicate that VP22 enhances the efficiency of the suicide gene transfer, thereby increases the cell-killing effect on tumor in vivo. PMID- 12885363 TI - [Study on transfer of interleukin-7 gene into ovarian carcinoma cells in vitro]. AB - OBJECTIVE: To investigate proliferative activity, expression of cell surface antigens and cytokines of ovarian carcinoma cells transferred by interleukin-7 (IL-7) gene, and its cytotoxic sensitivity to lymphokine activated killer (LAK) cells in vitro through gene transfection technique. METHODS: Ovarian carcinoma cell line SKOV3 were cultured and transferred by IL-7 cDNA. The proliferation activity of IL-7 gene transferred and nontransferred SKOV3 was observed with inverse phase-contrast light microscope, thiazoyl blue tetrazolium bromide assay and flow cytometry. The expressions of cell surface antigen human leucocyte antigen-ABC (HLA-ABC), human leucocyte antigen-DR (HLA-DR) and intercellular adhesion molecule-I (ICAM-I) were detected by indirect targeted FITC- flow cytometry. The secretions of IL-2, tumor necrosis factor alpha (TNF-alpha) and transforming growth factor (TGF-beta(1)) were assayed by enzyme linked immunosorbent assay. The cytotoxic sensitivity against LAK was detected by LDH release method. RESULTS: The morphology, proliferation, cell cycle distributions and HLA-ABC, HLA-DR expression of IL-7 gene transferred SKOV3 didn't change after IL-7 gene transfer. The expression of ICAM-I in IL-7 gene transferred SKOV3 significantly increased and TGF-beta(1) secretion of IL-7 gene transferred SKOV3 significantly decreased. The cytotoxic sensitivity of IL-7 gene transferred SKOV3 to LAK significantly elevated. CONCLUSION: Ovarian carcinoma cell line SKOV3, after transferred by IL-7 gene, remains unchanged proliferation and apoptosis, upregulation of ICAM-I, downregulation of TGF-beta(1) and increases the cytotoxic sensitivity to LAK in vitro. PMID- 12885364 TI - [Expression of survivin gene and its relation with the expression of bcl-2 and bax protein in epithelial ovarian cancer]. AB - OBJECTIVE: To study the expression of apoptosis related gene survivin and its relation with expression of bcl-2, bax protein in epithelial ovarian cancer. METHODS: Expression of survivin gene was evaluated by reverse transcription polymerase chain reaction (RT-PCR) in 35 cases of epithelial ovarian cancer, and compared with that in 10 cases of borderline cancer, 10 cases of benign tumors and 10 cases of normal tissue. Expression of bcl-2 and bax was detected by immunohistochemistry streptomycin-avidin-biotin-peroxidase complex (SABC) method, and correlation between them was analyzed. RESULTS: Expression of survivin gene was detected in a significantly greater proportion in epithelial ovarian cancer and borderline tumors than benign tumors and normal tissue. There was no relationship between survivin gene expression and FIGO stage, histologic grade, pathological type and lymphatic metastasis. Expression of bcl-2 and bax protein was positively and negatively correlated with expression of survivin gene respectively. CONCLUSION: survivin gene may play an important role in pathogenesis of ovarian cancer, apoptosis related gene bcl-2 may have a synergic role and bax have an antagonistic role with survivin gene in formation and progression of ovarian cancer. PMID- 12885365 TI - [Study on the expression of the gene deleted in colorectal carcinoma in ovarian carcinoma]. AB - OBJECTIVE: The purpose of this study was to explore the relationship between the loss of the gene deleted in colorectal carcinoma (DCC) gene expression in ovarian carcinoma and the transformation, progression of the tumor and its clinicopathological factors. METHODS: DCC gene mRNA expression were examined by reverse transcription polymerase chain reaction (RT-PCR) in 34 malignant, 10 benign and 10 normal ovarian samples. To clarify the expression of DCC gene by the DNA cloning and the DNA sequencing analysis in normal ovarian sample. RESULTS: The expression of DCC gene was lost in no normal ovarian tissues, in 2 (2/10) benign lesions, while the loss of DCC gene expression was found in 19 (19/34, 56%) carcinomas (P < 0.05). Similar findings were also noted when subdivision was into serous (14/18) and mucinous (4/11) categories (P < 0.05). The percentage of the loss of DCC gene expression in stage III - IV was 68%, in stage I - II was 22%, as well as 42% in grade I - II and 90% in grade III. 61% of the carcinoma with metastasis presented the loss of DCC gene expression, then there was 33% in which without metastasis. CONCLUSIONS: In ovarian carcinomas, loss of DCC gene expression was significantly related to the serous phenotype, a high histological grade and a more advanced stage. The results indicated that loss of DCC gene expression may play an important role in ovarian carcinomas and its progression. PMID- 12885366 TI - [Clinical evaluation of propess for induction of term pregnancy]. AB - OBJECTIVE: To explore the efficacy and safety of continuously released prostaglandin E(2) (PGE(2)) suppository-propess used for induction of term pregnancy. METHODS: A multicenter, prospective, case control clinical study was carried out, propess was used in 100 cases as study group, the suppository without PGE(2) was used in 49 cases as control group. The cervical maturity (by Bishop scoring), the time to labor starting, membrane rupture and delivery, the application of oxytocin, ceserean section rate, fetal and neonatal condition were compared between 2 groups after inserting of the suppository. At the same time, side effects caused by propess were investigated. RESULTS: Bishop score was increased >or= 2 points in 93% cases, >or= 3 points in 87% cases in study group, whereas only 4% cases whose Bishop score increased >or= 2 points in control group. The time to labor starting, membrane rupture, and delivery was shortened obviously in study group than that in control group after inserting suppository. The application of oxytocin was much less in study group, cesarean section rate was reduced in study group (32% vs 61%). There was no significant difference between 2 groups in fetal and neonatal conditions. The overstimulation of uterine contraction and mild gastrointestinal tract reaction occurred in 3 cases and 2 cases respectively in study groups. CONCLUSION: Propess can be used for induction of term pregnancy effectively and safely. PMID- 12885367 TI - [Observation of course of placental separation by continuous dynamic ultrasonography during third stage of labor]. AB - OBJECTIVE: To investigate the physiologic course of third stage of labor, and to judge the optimal time of natural placental separation permitted. METHODS: To observe and record the course of third stage of labor by continuous dynamic ultrasonography and to estimate the total amount of blood loss from placental separation within two hours after delivery by the method of weighing lost blood. RESULTS: Third stage of labor can be divided into four phases: the latent phase (4.37 +/- 3.78) min, the contraction phase (1.48 +/- 0.97) min, the detachment phase (0.50 +/- 0.00) min, and the expulsion phase (0.62 +/- 2.23) min. In 85% of the puerperas the third stage of labor takes about 10 minutes (average 6.94 minutes). The amount of postpartum hemorrhage and the risk of the patient increased prominently if the time of stage of labor was longer than 10 minutes (P < 0.01). CONCLUSION: The third stage of labor must be limited in 10 minutes to prevent postpartum hemorrhage. PMID- 12885368 TI - [Prevalence of postpartum thyroiditis in three different iodine intake areas]. AB - OBJECTIVE: To investigate the prevalence of postpartum thyroiditis (PPT) in three different iodine intake areas, to explore the relationship between iodine intake and PPT. METHODS: Panshan, Zhangwu and Huanghua are three different iodine intake areas. The median urinary iodine concentration were 103 micro g/L, 374 micro g/L and 614 micro g/L, respectively. One hundred and nineteen lactational women were investigated during the first year postpartum. Thyroid stimulating hormone (TSH), free thyroxine (FT(4)), free triiodothyronine (FT(3)), thyroid peroxidase antibody (TPOAb) and thyroglobulin antibody (TGAb) were determined with immulite method, and UI was determined by As-Ce(4+) catalytic spectrophotometry method. RESULTS: The prevalence of PPT is 11.8%, 10.1% of subclinical PPT, whereas 1.7% of clinical PPT. There was statistically significant difference in the frequency of PPT in three areas (Panshan 5.6%, Zhangwu 23.1% and Huanghua 6.8%, P < 0.05), but no difference in the frequency of thyroid autoimmune antibody (TAA). 50% of positive TAA women developed into PPT. CONCLUSION: Higher prevalence of subclinical PPT was found in the area with moderate iodine deficiency after iodine supplementation, suggesting a possible role of iodine supplementation in PPT. PMID- 12885369 TI - [Women's postpartum sexuality and delivery types]. AB - OBJECTIVE: To investigate the impact of delivery on the sexuality of primiparous women in China, and the association with delivery type. METHODS: We inquired 460 of primiparous women delivering of a live birth at the first affiliated hospital of Chongqing medical university from November 1, 2000 to July 31, 2001. It was a cross-sectional study using obstetric records and postal questionnaire survey by outpatients consultation six months after delivery. RESULTS: Ninety-four point seven percent had resumed sexual activity within six months of the birth. In the first three months after delivery, 70.6% of women experienced sexual problems, it declined to 34.2% at six months, but can not reaching the pre-pregnancy levels. There was no close relation between sexuality postpartum and delivery. (sexual satisfactory, desire, active rate, dyspareunia and pubococcygeal muscle intensity, P > 0.05). CONCLUSIONS: Postpartum sexuality was not significantly associated with delivery types. Sexual problems were very common after childbirth. More consultation and direction of postpartum sexuality recurrence is needed. PMID- 12885370 TI - [Applications of RHD zygosity test through polymerase chain reaction for prediction of fetus Rh D-positive phenotype]. AB - OBJECTIVE: To explore a molecular method for the determination of RHD zygosity. METHODS: Two pairs of primers were designed specific for downstream rhesus box and hybrid rhesus box according to the sequences in GenBank. Together with a pair of internal control primer, a dual-tube polymerase chain reaction (PCR) method was established for determination of the RHD zygosity. Thirty RhD-positive, D negative and weak D phenotype samples were evaluated by taking a recent restriction fragment length polymorphism (RFLP) method as reference. RESULTS: The results of the dual-tube PCR and RFLP are identical in 29 samples, which are also in concordance with the Rh phenotype, respectively. In one weak D sample, however, both methods tested as RHD(-)/RHD(-), which showed that there was one RHD(-) haplotype in this individual, but a false negative result in testing downstream rhesus box. CONCLUSIONS: The dual-tube PCR is a less-labored method for RHD zygosity detection. It can be used clinically in prenatal test to determine RHD zygosity of pregnant women's partner for prediction of fetus RhD phenotype. PMID- 12885371 TI - [A randomised clinical trial to study the effects of raloxifene hydrochloride on bone mineral density, biochemical markers of bone metabolism and serum lipids in postmenopausal women]. AB - OBJECTIVE: To determine the effect of raloxifene hydrochloride (RLX) on bone mineral density (BMD), biochemical markers of bone metabolism and lipid metabolism in Chinese postmenopausal women. METHODS: This was a multicenter, randomized, double blind placebo controlled study in China with a total of 204 postmenopausal women [mean age (60 +/- 5) years (x +/- s) and weight (63 +/- 9) kg (x +/- s)] treated with either RLX 60 mg (n = 102) or placebo (n = 102) daily for 12 months. BMD, serum lipid and bone markers were determined before and after drug administration. RESULTS: Compared to placebo, RLX produced a significant increase in both total lumbar spine and total hip BMD. For the lumbar spine, percentage increase in total BMD was 2.30% with RLX compared to a decrease of 0.08% with placebo (P < 0.001). Corresponding values for total hip BMD were 2.46% increase for RLX and 1.07% for placebo (P < 0.05). For biochemical markers of bone metabolism, serum osteocalcin and C-telopeptide, percentage decrease were 27.6% and 24.0% in raloxifene-treated subjects. Corresponding values in placebo were 10.6% decrease and 15.8% increase (RLX compared to placebo, both P < 0.001). For total cholesterol and low-density lipoprotein cholesterol, percentage decrease were 6.4% and 34.6% in the raloxifene-treated group. Corresponding values in placebo were 1.4% increase and 19.1% decrease (RLX compared to placebo, both P < 0.001). No differences were found for high-density lipoprotein cholesterol or triglyceride levels between the two groups. Only 5 subjects discontinued early due to an adverse event (3 in the RLX group and 2 in the placebo group). CONCLUSIONS: This study confirms that RLX exerts positive effects on the skeleton, increasing BMD and decreasing biochemical markers of bone metabolism, and decreased total cholesterol and low-density lipoprotein cholesterol in postmenopausal women in China. PMID- 12885372 TI - [In vitro maturation, fertilization and embryo transfer of human immature oocyte]. AB - OBJECTIVE: To try a new method for treating infertile women by in vitro maturation technique. METHODS: To set up in vitro maturation (IVM) program, including stimulating regimen, oocyte retrieval technique, and culture system of the immature oocytes with the medium consisted of essential supplements and human mature follicular fluid. IVM was performed in 30 infertile patients (35 cycles), (1) with polycystic ovary syndrome (POCS) (14/30), (2) ovary hyperstimulation syndrome (OHSS) in the previous stimulating cycles (6/30), and (3) retardation of the oocyte growth in conventional IVF cycles (10/30). RESULTS: Totally 203 oocytes were obtained, with a mean of 5.8 oocytes each cycle. First polar bodies were observed in 156 oocytes, maturation rate was 76.8%. There were 120 oocytes getting into pronucleus stage, normal fertilization rate was 76.9%. Embryo transfer was performed in 33 cycles. Eight pregnancies were established, pregnancy rate was 24%. Five patients have delivered 7 healthy babies. CONCLUSIONS: The result of our study indicated that IVM is one of the new assisted reproductive technologies for the infertile women with PCOS, poor response to stimulation and the history of OHSS. Human mature follicular fluid would be an ideal component in the maturation medium for oocyte maturation in vitro and provides a much simplified and practical method for clinical use. PMID- 12885373 TI - Attention and scintillation. AB - The scintillating grid is a recently discovered visual illusion, created by superimposing white discs on the intersections of orthogonal gray bars on a black background. Even though the entire display is stationary, observers perceive dark spots appearing and disappearing rapidly within some of the white discs. This scintillation effect is correlated with eye position and eye movements. Here we investigate whether covert shifts of attention, as revealed by cueing and visual search paradigms, can also affect the illusion. We find that the chance of a particular intersection scintillating is directly correlated with distance from the attentional focus, regardless of the location of the fixation point. This suggests that the dynamics of this illusion might reflect the distribution of attention in space and time. PMID- 12885374 TI - Orientation opponency in human vision revealed by energy-frequency analysis. AB - Studies of second-order visual processing have primarily been concerned with understanding the mechanisms for detecting spatiotemporal variations in such attributes as contrast, orientation, spatial frequency, etc. Here, we have examined the orientation characteristics of second-order processes using bandpass noise whose Fourier energy is sinusoidally modulated across orientation, rather than across space or time. Sensitivity for detecting orientation-energy modulations was measured as a function of modulation frequency. The sensitivity function was bandpass, with a pronounced peak at an orientation frequency of 4 cycles/pi. An inverse Fourier transform of the sensitivity function revealed a filter profile displaying a centre-surround antagonism across orientation, with an excitatory centre within 6-9 deg and inhibitory lobes at 15-20 deg from the filter's centre. The degree of centre-surround antagonism increased with stimulus size far beyond the spatial range of the first-order filters (more than 64 times the dominant spatial wavelength of the noise carrier). These results suggest that second-order processing involves 'orientation-opponent' channels that extract differences in first-order outputs across orientation over a wide area of the visual field. PMID- 12885375 TI - Temporal characteristics of binocular rivalry: visual field asymmetries. AB - Very little is known about the mechanisms that drive the alternation between the two views during binocular rivalry. A key property of the rivalry process is the rate at which the two views alternate. Understanding the factors that affect the rate of the alternation is critical to the final understanding of the underlying process. Using a circular and a radial grating as the rivalry stimuli, we observed a significantly faster binocular rivalry when stimuli were presented in the right visual field than that in the left visual field for the right-handed observers, and a reversed asymmetry for the left-handed observers. In both groups, rivalry was faster for stimuli presented in the lower visual field than that in the upper visual field. This pattern of results suggests that (1) rivalry is likely a locally driven process and (2) the visual brain in the left hemisphere may be the faster one of the two hemispheres in right-handed people. PMID- 12885376 TI - Divided attention and visual search for simple versus complex features. AB - Under what search conditions does attention affect perceptual processes, resulting in capacity limitations, rather than affecting noisy decision-making processes? Does parallel or serial processing cause the capacity limitations? To address these issues, we varied stimulus complexity, set size, and whether distractors were mirror images of the target. Both target detection and localization produced similar patterns of results. Capacity limitations only occurred for complex stimuli used in within-object conjunction searches. Parallel processing, rather than serial processing, probably caused these capacity limitations. Moreover, although mirror-image symmetry adversely affected early visual processing, it did not place additional demands on attention. PMID- 12885377 TI - "Phase capture" in the perception of interpolated shape: cue combination and the influence function. AB - This study was concerned with what stimulus information observers use to judge the shape of simple objects. We used a string of four Gabor patches to define a contour. A fifth, center patch served as a test pattern. The observers' task was to judge the location of the test pattern relative to the contour. The contour was either a straight line, or an arc with positive or negative curvature (the radius of curvature was either 2 or 6 deg). We asked whether phase shifts in the inner or outer pairs of patches distributed along the contour influence the perceived shape. That is, we measured the phase shift influence function. We found that shifting the inner patches of the string by 0.25 cycle results in almost complete phase capture (attraction) at the smallest separation (2 lambda), and the capture effect falls off rapidly with separation. A 0.25 cycle shift of the outer pair of patches has a much smaller effect, in the opposite direction (repulsion). In our experiments, the contour is defined by two cues--the cue provided by the Gabor carrier (the 'feature' cue) and that defined by the Gaussian envelope (the 'envelope' cue). Our phase shift influence function can be thought of as a cue combination task. An ideal observer would weight the cues by the inverse variance of the two cues. The variance in each of these cues predicts the main features of our results quite accurately. PMID- 12885378 TI - A paradox of temporal perception revealed by a stimulus oscillating in colour and orientation. AB - Psychophysical experiments with stimuli oscillating concurrently in colour and orientation revealed an apparently paradoxical dissociation between the perceived simultaneity of stimulus changes and the perceptual pairing of the events demarked by those changes. When subjects were required to report whether changes in colour and orientation were simultaneous, judgements were generally accurate within +/-10 ms. When subjects were required to report which colour was paired predominantly with which orientation, judgements showed a systematic temporal bias of up to 50 ms in favour of colour. This dissociation between different temporal judgements concerning the same stimulus sequence is not predicted by any of the current models of binding in conscious vision. We propose an account of these data based on the temporal response properties of colour- and orientation selective model neurons such that the perceived pairing of visual attributes is modelled as the cross-correlation of time-varying neural response profiles and thus reflects both neuronal latencies and the rate of rapid adaptation rather than simply the temporal pattern of responses to stimulus transitions. PMID- 12885379 TI - Border distinctness in amblyopia. AB - On the basis of the contrast sensitivity loss in amblyopia which mainly affects higher spatial frequencies, one would expect amblyopes to perceive sharp edges as blurred. We show that they perceive sharp edges as sharp and have veridical edge blur perception. Contrary to the currently accepted view, this suggests that the amblyopic visual system is not characterized by a blurred visual representation. PMID- 12885380 TI - Human visual object categorization can be described by models with low memory capacity. AB - Studies of high-level models of visual object categorization have left unresolved issues of neurobiological relevance, including how features are extracted from the image and the role played by memory capacity in categorization performance. We compared the ability of a comprehensive set of models to match the categorization performance of human observers while explicitly accounting for the models' numbers of free parameters. The most successful models did not require a large memory capacity, suggesting that a sparse, abstracted representation of category properties may underlie categorization performance. This type of representation--different from classical prototype abstraction--could also be extracted directly from two-dimensional images via a biologically plausible early vision model, rather than relying on experimenter-imposed features. PMID- 12885381 TI - Physical stability of a microcrystalline beta-sitosterol suspension in oil. AB - Sterols have been shown to reduce plasma cholesterol by blocking the absorption of cholesterol from the gut. The physical properties of crystalline plant sterols limit their use in foods. A coarse-grained structure can be avoided by recrystallisation, a method that affords a reduction in the particle size. A previous work described how to produce a microcrystalline beta-sitosterol suspension. The present study deals with the stability of that suspension. Recrystallisation was carried out by two different methods; one based on rapid the other based on slow cooling, whereby six different compositions were made containing 5-30% of beta-sitosterol and secondly either 5 or 20% water was added. The particle size and habit were evaluated during a 16 weeks storage period (+4 or -19 degrees C) by way of optical microscopy. The crystal structure and degree of crystallinity was analysed by X-ray diffraction. Suspensions can, in most cases, be stored for 16 weeks without any changes to the size and habit. The only evidence of crystal growth came from a suspension with a low sterol concentration at a temperature of +4 degrees C. This is due to the dissolution-diffusion process which is affected by temperature and viscosity. Suspensions containing higher amounts of sterol remained stable, if stored at +4 or -19 degrees C, for 16 weeks. The suspensions included both hemihydrous and monohydrous beta sitosterol crystals. Suspensions containing less sterol showed greater amounts of monohydrated crystals. This illustrates more water penetration into the crystals. A higher sterol concentration led to a larger number of smaller crystals creating reflections similar to hemihydrated crystals. PMID- 12885382 TI - Effect of colloidal carriers on ascorbyl palmitate stability. AB - Active compounds can be protected against degradation by incorporation into colloidal carrier systems. The stabilizing effect of carrier systems for ascorbyl palmitate (AP) was investigated using microemulsions (ME), liposomes and solid lipid nanoparticles (SLN). Analysis of chemical stability by HPLC showed that AP is most resistant against oxidation in non-hydrogenated soybean lecithin liposomes, followed by SLN, w/o and o/w ME, and hydrogenated soybean lecithin liposomes. The molecular environment of the AP-like nitroxide probe (C(16)-Tempo) in colloidal carriers was characterized using electron paramagnetic resonance (EPR) spectroscopy. We have found that the nitroxide groups are located in environments with different polarity and mobility. The hydrophilic part of AP is the reactive moiety, and high stability is obtained in systems in which this part is exposed to a less polar environment. Additionally, the determined accessibility of nitroxide groups to reduction correlated well with the chemical stability of AP. It is more deeply immersed in the interface when entrapped in a liquid-state carrier than when applied in gel-state particles. Encapsulation of AP in SLN core leads to greater stability. We conclude that the location of the sensitive group of the drug-molecule in a carrier system is crucial for its stability. PMID- 12885383 TI - Formulation and characterization of DNA-polyethylenimine-dextran sulfate nanoparticles. AB - Polyethylenimine (PEI) is a promising non-viral gene delivery polymer that produces high transfection efficiency both in vitro and in vivo. The use of PEI, however, is hindered by its toxicity, reflecting its polycationic nature. In an attempt to decrease this charge-dependent cytotoxicity, a polyanionic polymer, dextran sulfate (DS), has been incorporated into self-assembling PEI-DNA complexes with zinc as stabilizing agent. Spherical particles with a mean particle size of approximately 200 nm and a polydispersity index of 0.2 were achieved using the following optimal conditions: PEI solutions at pH 8, PEI/DS mass ratios of >or=2, and 25 microM zinc sulfate. Plasmid DNA was completely condensed within the nanoparticles as confirmed by an ethidium bromide accessibility assay. This result correlates well with DNase protection studies which find partial protection of the DNA nanoparticles from degradation by the enzyme. The DNA was incorporated into the PEI-DS particles with a high efficiency (>95%) and maintained a primarily supercoiled B-form as determined by gel electrophoresis and circular dichroism. The cytotoxicity of the DNA nanoparticles appeared to decrease as the amount of DS in the formulation was increased and they produced moderate transfection activities that were only modestly inhibited by the presence of serum. PMID- 12885384 TI - Assessment of diclofenac permeation with different formulations: anti inflammatory study of a selected formula. AB - The aim of this study was to improve the transdermal permeation of sodium diclofenac. Permeation studies were carried out in vitro using human skin (0.4 mm thick) from plastic surgery as a membrane. Four liquid formulations of 1% (w/w) sodium diclofenac were assayed: three ternary solvent systems (M4, M5, M6) and one microemulsion (M3). A 1% (w/w) solution of sodium diclofenac and a commercially available semisolid preparation were tested as reference formulations. The following permeation parameters for diclofenac were assessed: permeability coefficient, flux and drug permeated and retained in the skin at 24 h. The highest values of these parameters were obtained with formula M4, which contains transcutol 59.2%, oleic acid 14.9% and d-limonene 5% (w/w) as permeation enhancers. The anti-inflammatory activity of this formula was compared with that of the semisolid preparation on carrageenan-induced paw edema in rats. As expected from in vitro results, the M4 diclofenac delivery system showed higher activity than the semisolid preparation, both when applied locally (to the inflammation area) and when applied systemically (to the back). Neither treatment irritated the skin when tested on rabbits in a 72-h trial. These results suggest that topical delivery of sodium diclofenac with an absorption enhancer such as a mixture of oleic acid and d-limonene (M4) may be an effective medication for both dermal and subdermal injuries. PMID- 12885385 TI - Characterisation and deposition studies of engineered lactose crystals with potential for use as a carrier for aerosolised salbutamol sulfate from dry powder inhalers. AB - Lactose particles with different elongation ratio, roundness, polymorphic form and crystallinity were prepared by a one-step crystallisation process using varying ratios of acetone/water. The crystals were characterised using image analysis optical microscopy, scanning electron microscopy, differential scanning calorimetry and X-ray powder diffraction. The elongation ratio was found to increase with increasing acetone ratio which therefore, appears to accelerate the growth in length rather than width and/or thickness. The crystallinity and polymorphic forms were also acetone-concentration dependent. For example, the crystals formed using 65-80% v/v acetone were almost all of the alpha-form whereas at 85% v/v a small amount of beta-form was precipitated, as detected by a peak at the reflection angle 2 theta=10.4 in the X-ray diffractogram. When 90% v/v acetone was incorporated a mixture of alpha- and beta-forms were produced in almost equal quantity, whereas, with 95% v/v acetone the beta-form predominated. At high acetone concentration (90 and 95% v/v), the crystallisation proceeded rapidly leading to the creation of some amorphous content. The 63-90-microm sieve cut of either commercial grade lactose (CL) or crystallised lactose was mixed with salbutamol sulfate and dispersibility was determined using the twin stage liquid impinger. All the formulations containing carrier particles generated by crystallization from solvent showed higher dispersibility and fine particle fraction (FPF) of the drug compared to the formulation made containing CL. The carrier that showed the highest elongation ratio (produced from an 85% acetone 15% water solution), when mixed with salbutamol sulfate produced the highest dispersibility (38.5%) and highest FPF (29.24%). These parameters were six times higher than the values obtained with the formulation containing CL. PMID- 12885386 TI - Inhibition of mevalonate pathway is involved in alendronate-induced cell growth inhibition, but not in cytokine secretion from macrophages in vitro. AB - Bisphosphonates are antiresorptive drugs used for the treatment of metabolic bone diseases. They can be divided into two different pharmacological classes: nitrogen-containing and non-nitrogen-containing bisphosphonates. Non-nitrogen containing bisphosphonates, like clodronate, are metabolised to a toxic ATP analogue preventing osteoclast mediated bone resorption. Nitrogen-containing bisphosphonates, including alendronate, prevent osteoclast function by inhibiting the mevalonate pathway. Clodronate is known to have anti-inflammatory properties while alendronate induces cytokine secretion from lipopolysaccharide- (LPS) induced macrophages. This study investigates whether the cytotoxicity and cytokine production induced by alendronate and LPS could be counteracted by clodronate or products of mevalonate pathway: oxidized low density lipoprotein (ox-LDL), farnesol and geranylgeraniol. Treatment with alendronate increased LPS induced secretion of IL-1beta, IL-6 and TNF-alpha from RAW 264 macrophages 2.4-, 1.4- and 1.8-fold, respectively. This treatment was cytotoxic for macrophages as indicated by lowered cell viability. Clodronate and ox-LDL both counteracted the cytokine secretion and cytotoxicity of alendronate. Farnesol and geranylgeraniol did neither reverse the cytokine secretion nor reduce the cytotoxicity of alendronate. Clodronate and ox-LDL were able to counteract the effects of alendronate on macrophages in vitro, probably by their known ability to inhibit DNA binding activity of transcription factors, nuclear factor-kappaB (NF-kappaB) and activating protein-1 (AP-1). These findings suggest that inhibition of mevalonate pathway is not the mechanism responsible for the proinflammatory response caused by alendronate, as it is in alendronate-induced apoptosis and prevention of osteoclast function. PMID- 12885387 TI - The application of the convective diffusion model and the film equilibrium model to surfactant-facilitated dissolution of gliclazide. AB - Gliclazide is practically insoluble in water, and has low dissolution rate. Therefore, it was of interest to improve its dissolution rate using anionic and cationic surfactants. The intrinsic dissolution rates of gliclazide in solutions of sodium dodecyl sulfate (SDS) and in solutions of tetradecyltrimethyl ammonium bromide (TDTMAB) were measured using the rotating disk method to study the convective diffusion transport of drug-loaded micelles. Two different approaches were applied to the experimental data; the convective diffusion model and the film equilibrium model. The two approaches are based on the same fundamental assumptions differing only in their interpretation of the diffusional boundary layer. The results obtained from the film equilibrium model were less satisfactory, and in case of TDTMAB the model was inapplicable (negative diffusion coefficient). While excellent results were obtained from the convective diffusion model. The free solute diffusion coefficient (D(s)) obtained experimentally was 2.47 x 10(-5) cm(2)/s, and the diffusion coefficient of the drug-loaded SDS micelle (D(sm)) estimated was 1.74 x 10(-6) cm(2)/s. The drug loaded SDS micelle radius was 14 A. The thickness of the diffusional boundary layer was 54 and 22 microm for the free solute and the drug-loaded SDS micelle, respectively. TDTMAB showed lower effect in improving the dissolution rate of gliclazide than SDS. The drug-loaded TDTMAB micelle diffusion coefficient was 1.03 x 10(-6) cm(2)/s. The radius of the drug-loaded TDTMAB micelle and the boundary layer thickness were 24 A and 19 microm, respectively. PMID- 12885388 TI - Differential scanning calorimetry differences in micronized and unmicronized nimesulide uptake processes in biomembrane models. AB - Nimesulide release from micronized and unmicronized drug particles was tested at pH 7.4 by measuring the transfer to dimyristoylphosphatidylcholine liposomes (multilamellar and unilamellar vesicles), chosen as a biomembrane model. The perturbing effect of increasing molar fractions of pure nimesulide on the thermotropic behaviour of dimyristoylphosphatidylcholine liposomes was investigated by differential scanning calorimetry. In order to study the drug dissolution process by its uptake into void liposomes, measurements were carried out on suspensions of blank liposomes added to weighed amounts of free powdered nimesulide (micronized and unmicronized). The amount of drug transferred was quantified by comparing the effect caused by the dissolved and released drug to that caused by the free drug that had been previously molecularly dissolved in the liposomes. The calorimetric results show that the dissolution rate depends on the nimesulide form (micronized or unmicronized), and that the transfer to the void liposomes is quicker when the drug is in a micronized form. The uptake was faster when unilamellar vesicles were used instead of multilamellar vesicles because of the greater lipid surface. The calorimetric technique could represent an alternative 'in vitro' method that can be applied to the study of the dissolution kinetics directly at the site of drug uptake, mimicking a biological system. PMID- 12885389 TI - Endospores of B subtilis are pyrogenic and activate Mono Mac 6 cells: importance of the CD14 receptor. AB - The monocytic cell line Mono Mac 6 is sensitive to pyrogens and interleukin-6 secretion is induced after exposure to pyrogens. The aim of this study is to examine the pyrogenic activity and the interleukin-6-inducing capacity of the Gram-positive B. subtilis bacteria, endospores and isolated cell wall components. Furthermore the involvement of CD14 in activation of interleukin-6 release is investigated. All test substances are pyrogenic in the rabbit pyrogen test. The test substance is incubated with monocytic cells (Mono Mac 6) for 24 h and the secreted interleukin-6 is determined in a sandwich immunoassay. B. subtilis bacteria and endospores induce interleukin-6 in a dose-dependent manner. Endospores are less potent than bacteria. Lipoteichoic acid (LTA) isolated from B. subtilis induces interleukin-6 in a dose-dependent manner, whereas muramyl dipeptide (MDP) is unable to induce interleukin-6. Lipopolysaccharides (LPS) dose dependently induce interleukin-6 release, but the curve differs from that of LTA both in shape and offset. The interleukin-6 secretion induced by LPS, LTA and B. subtilis bacteria can be blocked by 73-85% by an antibody directed against CD14, whereas the antibody only blocks 25% of B. subtilis endospores-induced interleukin-6 release. The results might indicate that B. subtilis endospores use an additional pathway to CD14 to activate mononuclear cells. PMID- 12885390 TI - Development of sesquiterpenes from Alpinia oxyphylla as novel skin permeation enhancers. AB - To improve the drug permeation into and/or across the skin, essential oils extracted from Alpinia oxyphylla (AO) were evaluated using in vitro and in vivo permeation techniques with Wistar rats as the animal model. Hydrocarbons and oxygenated sesquiterpenes were the major components in the lower-polarity fraction (AO-1) and higher-polarity fraction (AO-2), respectively. Permeation of indomethacin was significantly enhanced after treatment with AO-1 and AO-2 in the in vitro and in vivo studies. AO-2 generally showed a higher ability to promote drug permeation compared to AO-1. The increment of skin/vehicle partitioning may be the predominant mechanism for this enhancing activity. Both transepidermal water loss (TEWL) and colorimetric evaluation showed limited irritation to skin by AO essential oils at the macroscopic level. Human skin fibroblasts were used to investigate the in vitro screening of skin toxicity. AO-1 slightly increased prostaglandin E(2) (PGE(2)) formation from skin fibroblasts. A striking result was observed with AO-2, which greatly inhibited the release of PGE(2). Moreover, both AO essential oils had no statistically significant effect on PGE(2) release by human lung epithelial cells. The results of this study indicate that skin disruption and inflammation do not necessary correspond to the enhancing efficiency of the enhancers tested. PMID- 12885391 TI - Assessment of drug salt release from solutions, suspensions and in situ suspensions using a rotating dialysis cell. AB - A rotating dialysis cell consisting of a small (10 ml) and a large compartment (1000 ml) was used to study the release of drug salt (bupivacaine 9-anthracene carboxylate) from (i). solutions, (ii). suspensions and (iii). in situ formed suspensions. Initial release experiments from suspensions indicated that the release of drug salt in deionized water was predominantly limited by the diffusion across the membrane whereas it is essentially dissolution rate controlled in 0.05 M phosphate buffer (pH 7.40). Thus, the in vitro model appears to have a potential in formulation screening when phosphate buffer is used as release media. Generally, the initial release of the drug salt from in situ suspensions occurred faster as compared to conventional suspensions, probably due to incomplete precipitation of the drug salt, and hence formation of supersaturated solutions where the rate of release is predominantly determined by the concentration gradient. However, when an adequately concentrated solution of the drug salt was used to prepare the in situ suspension, the initial fast release was followed by a substantial sustained release indicating that the release had become dissolution rate limited. PMID- 12885392 TI - Spray-dried redispersible oil-in-water emulsion to improve oral bioavailability of poorly soluble drugs. AB - A physically stabilized dry emulsion dosage form reforming the original emulsion after rehydration was developed by spray-drying a liquid oil-in-water emulsion containing maltodextrin as carrier and sodium caseinate as emulsifying agent. Several oil:water as well as maltodextrin:water ratios were tested, the homogenization and spray-drying processes and the reconstitution properties were investigated and an optimum formulation was selected for poorly soluble drug incorporation, having an identical oil:water and carrier:water ratio of 10% (w/w) and a load of solid material of 20% (w/w). Lipophilic 5-phenyl-1,2-dithiole-3 thione (5-PDTT) was selected as a model drug. 5-PDTT release from the solid state emulsion was studied using an in vitro two-phase stirred model and the relative bioavailability of 5-PDTT in the dry emulsion was obtained in the rabbit after oral administration of the reconstituted emulsion, compared to a 5-PDTT sulfobutyl ether 7 beta-cyclodextrin complex in solution. Incorporation of 5-PDTT in the oil phase neither affects the surface morphology of the powder nor the reconstitution, the droplet size or the drug releasing properties and, furthermore, allows a 3-fold improvement of 5-PDTT relative bioavailability in rabbit after oral administration. These results indicate that dry emulsions may be considered as relevant dosage forms to improve bioavailability of poorly absorbable lipophilic drugs. PMID- 12885393 TI - How do roll compaction/dry granulation affect the tableting behaviour of inorganic materials? Comparison of four magnesium carbonates. AB - The effect of roll compaction/dry granulation on the particle and bulk material characteristics of different magnesium carbonates was evaluated. The flowability of all materials could be improved, even by the application of low specific compaction forces. The tablet properties made of powder and dry granulated magnesium carbonate were compared. Roll compaction/dry granulation resulted in a modified compactibility of the material and, consequently, tablets with reduced tensile strength. The higher relative tap density of the compacted material does not allow a densification to the same extent as the uncompacted powder. The degree of densification during tableting can be expressed as the ratio of the relative tablet density to the relative tap density of the feed material. Increasing the specific compaction forces resulted in higher apparent mean yield pressure, gained from Heckel plots, of all materials analysed. The partial loss of compactibility leads to the demand of low loads during roll compaction. Comparing the tablet properties of different magnesium carbonates reveals an obvious capping disposition. However, it depends on the type of magnesium carbonate, the specific compaction force and also on the tableting force applied. PMID- 12885394 TI - Applications of a new device (spindle) for improved characterization of drug release (dissolution) of pharmaceutical products. AB - A crescent spindle (patent pending) is described which may be used in place of the USP paddle component in USP dissolution apparatus 2. The new spindle is curve shaped, corresponding to the bottom of a dissolution vessel, with attached bristles to fill in the gap between the spindle and the surface of the vessel. The geometry of the new spindle provides more efficient mixing than the USP paddle and prevents accumulation of disintegrated material (no cone formation). Using the new spindle, in comparison with the USP paddle, dissolution characteristics of three drug products: 250 mg amoxicillin capsules, 15.6 g acetylsalicylic acid (ASA) boluses and 200 mg carbamzepine tablets were evaluated. The experimental conditions for dissolution testing with the two stirring devices included; 900 ml of 0.05 M phosphate buffer, pH 6.8 with 50 rpm, 900 ml of 0.05 M acetate buffer, pH 4.5-ethanol (7:3) with 50 rpm, and water containing 1% sodium lauryl sulphate with 75 rpm for amoxicillin capsules, ASA boluses and carbamazepine tablets, respectively. Uncharacteristic of the test products, which are fast release, the USP paddle provides significantly slower drug release. For example, 90 min for <80% drug release vs. 10 min for >90% for amoxicillin capsules and 6 h for 80% vs. 30 min for >90% for ASA boluses with USP paddle vs. the new spindle. In case of the carbamazepine tablets, three products which are bioequivalent and prescribed interchangeably, the USP paddle method shows significantly different dissolution characteristics. However, with the new device, all these products show similar drug release characteristics, a better reflection of product release characteristics and in vivo drug release behaviour. Compared with the USP paddle, the suggested device (spindle) provides improved stirring and mixing which appears to provide more appropriate (biorelevant) characterization of pharmaceutical products. PMID- 12885395 TI - Oral bioavailability of the antioxidant astaxanthin in humans is enhanced by incorporation of lipid based formulations. AB - Astaxanthin is a carotenoid with antioxidant properties, synthesised by plants and algae, and distributed in marine seafood. Astaxanthin is also available as a food supplement, but, like other carotenoids, is a very lipophilic compound and has low oral bioavailability. However, bioavailability can be enhanced in the presence of fat. There is not much information in the literature about the pharmacokinetics of oral astaxanthin in humans. In this open parallel study, healthy male volunteers received a single dose of 40 mg astaxanthin, as lipid based formulations or as a commercially available food supplement, followed by blood sampling for further analysis of plasma concentrations. Pharmacokinetic parameters were calculated to evaluate the extent and rate of absorption from each formulation. The elimination half-life was 15.9+/-5.3 h (n=32), and showed a mono-phasic curve. Three lipid based formulations: long-chain triglyceride (palm oil) and polysorbate 80 (formulation A), glycerol mono- and dioleate and polysorbate 80 (formulation B), and glycerol mono- and dioleate, polysorbate 80 and sorbitan monooleate (formulation C), all showed enhanced bioavailability, ranging from 1.7 to 3.7 times that of the reference formulation. The highest bioavailability was observed with formulation B, containing a high content of the hydrophilic synthetic surfactant polysorbate 80. PMID- 12885396 TI - Biochemical and pharmacokinetic evaluation of a novel pyrimidine nucleoside nitric oxide donor as a potential anticancer/antiviral agent. AB - The objective of this study was to determine the physiochemical, biochemical and pharmacokinetic properties of 5-iodo-3'-O-nitro-2'-deoxyuridine (INUdR), a novel prodrug releasing both nitric oxide (NO) and 5-iodo-2'-deoxyuridine. The INUdR partition coefficient (log P=1.12) was determined by both the shake-flask method and by calculation using Interactive Analysis Log P Program. In vitro binding of INUdR to bovine serum albumin (BSA) was estimated using an ultrafiltration method to be 65 to 77%, depending on the INUdR concentration. INUdR was stable in phosphate buffer (pH 7.4) and in water, at both ambient temperature and at 37 degrees C. INUdR was resistant to phosphorolysis when incubated with thymidine phosphorylase. Plasma, L-cysteine and glutathione catalyzed release of NO from INUdR, as determined using the Griess reaction. In all three systems, the release of NO by INUdR was equal to or greater than that of the reference drug isosorbide dinitrate. The pharmacokinetics of INUdR following single intravenous bolus and oral doses of INUdR (40 mg/kg) to male Sprague-Dawley rats were characterized by a short elimination half-life (T(1/2) 0.27 h), a large steady-state volume of distribution (V(ss) 0.89 l/kg) and high oral bioavailability (F=0.95). In conclusion, INUdR lipophilicity, shelf-stability, and resistance towards catabolic breakdown by thymidine phosphorylase, together with its non spontaneous, yet considerable NO release, constitute favorable characteristics of a potential anticancer/antiviral agent. PMID- 12885397 TI - The cytochrome c3-[Fe]-hydrogenase electron-transfer complex: structural model by NMR restrained docking. AB - Cytochrome c(3) (M(r) 13000) is a low redox potential cytochrome specific of the anaerobic metabolism in sulfate-reducing bacteria. This tetrahemic cytochrome is an intermediate between the [Fe]-hydrogenase and the cytochrome Hmc in Desulfovibrio vulgaris Hildenborough strain. The present work describes the structural model of the cytochrome c(3)-[Fe]-hydrogenase complex obtained by nuclear magnetic resonance restrained docking. This model connects the distal cluster of the [Fe]-hydrogenase to heme 4 of the cytochrome, the same heme found in the interaction with cytochrome Hmc. This result gives evidence that cytochrome c(3) is an electron shuttle between the periplasmic hydrogenase and the Hmc membrane-bound complex. PMID- 12885398 TI - Transferrins selectively cause ion efflux through bacterial and artificial membranes. AB - Serum transferrin, ovotransferrin and lactoferrin constitute the most notable members of the transferrin family. Among their multiple biological functions, they possess an important antibacterial activity. These proteins can permeate the Escherichia coli outer membrane, reaching the inner membrane where they selectively cause permeation of ions, resulting in dissipation of the electrical potential without affecting the pH gradient. Similar results were obtained using artificial liposomes, suggesting a direct action of the proteins on the lipid bilayer, which was mediated by detectable conformational changes in their structures. PMID- 12885399 TI - Two distinct calcium-calmodulin interactions with N-terminal regions of the olfactory and rod cyclic nucleotide-gated channels characterized by NMR spectroscopy. AB - The interactions of calcium-calmodulin with two fragments of the N-terminal domains of the olfactory alpha-subunit and rod beta-subunit cyclic nucleotide gated channels have been investigated using nuclear magnetic resonance spectroscopy. The results indicate that in the two cases both the N-terminal and the C-terminal calmodulin lobes are involved in the interaction. The olfactory cyclic nucleotide-gated channel segment forms a 1:1 complex with calmodulin, whereas the rod fragment forms a 2:1 complex. The correlation times of the two complexes, as estimated by (15)N relaxation studies, are compatible with the observed stoichiometries. These results indicate differences in the mode of action by which calmodulin modulates the activity of both channels, and suggest either that the rod channel is modulated through a simultaneous interaction of two beta-subunits with calmodulin or that other regions of the N-terminus are necessarily implicated in the binding. PMID- 12885400 TI - Identification of the nuclear receptor CAR:HSP90 complex in mouse liver and recruitment of protein phosphatase 2A in response to phenobarbital. AB - The nuclear receptor CAR, a phenobarbital (PB)-responsive transcription factor, translocates into the nucleus of hepatocytes after PB induction. In non-induced mice, CAR forms a physical complex with heat shock protein 90 (HSP90) in the cytoplasm. In response to PB induction, protein phosphatase 2A is recruited to the CAR:HSP90 complex. This recruitment may lead CAR to translocate into the nucleus, consistent with the inhibitions of nuclear CAR accumulation in PB induced mouse primary hepatocytes by okadaic acid as well as by geldanamycin. PMID- 12885401 TI - Resistin is expressed in different rat tissues and is regulated in a tissue- and gender-specific manner. AB - Resistin is a polypeptide hormone first reported from human and rodent adipocytes. In order to better define the potential biological role of resistin we undertook a detailed analysis of its expression in different rat tissues. We demonstrate by real-time reverse transcription polymerase chain reaction, Southern blotting and immunohistochemistry that resistin is expressed not only in brown and white adipose tissue, but also in the stomach, small and large intestines, adrenal gland, and skeletal muscle. Food deprivation led to a decrease in resistin mRNA expression only in adipose tissue, not in any of the other tissues studied. Furthermore, resistin mRNA expression is higher in males than in females in adipose tissue, not in any of the other tissues. Thus, our data suggest that resistin is not exclusively localized in adipocytes, and indicate that its expression is regulated in a tissue- and sex-specific manner. PMID- 12885402 TI - Differential expression of sphingolipids in P-glycoprotein or multidrug resistance-related protein 1 expressing human neuroblastoma cell lines. AB - The sphingolipid composition and multidrug resistance status of three human neuroblastoma cell lines were established. SK-N-FI cells displayed high expression and functional (efflux) activity of P-glycoprotein, while multidrug resistance-related protein 1 was relatively abundant and most active in SK-N-AS cells. These two cell lines exhibited higher sphingolipid levels, compared to SK N-DZ, which had the lowest activity of either ATP-binding cassette transporter protein. SK-N-DZ cells also differed in ganglioside composition with predominant expression of b-series gangliosides. In conclusion, these three neuroblastoma cell lines offer a good model system to study sphingolipid metabolism in relation to ATP-binding cassette transporter protein function. PMID- 12885403 TI - A gene-specific effect of an internal deletion in the Bdp1 subunit of the RNA polymerase III transcription initiation factor TFIIIB. AB - The Saccharomyces cerevisiae RPR1 gene encodes the RNA subunit of its RNase P, which processes RNA polymerase (pol) III primary transcripts. RPR1, which is transcribed by pol III, has been isolated as a multicopy suppressor of a specific small internal deletion (amino acids 253-269) in the Bdp1 subunit of transcription factor TFIIIB, the core pol III transcription factor. The selective effect of this Bdp1 deletion on RPR1 transcription has been analyzed in vitro. It is shown that TFIIIC-dependent assembly of TFIIIB on the RPR1 promoter is specifically sensitive to this Bdp1 deletion, leading to gene-specifically defective single-round and multiple-round transcription. PMID- 12885404 TI - Human immunodeficiency virus-1 Nef protein interacts with Tat and enhances HIV-1 gene expression. AB - The human immunodeficiency virus (HIV-1) Nef protein is now regarded as a regulatory protein responsible not only for establishment of infection and increased pathogenesis but also for enhancement of viral replication. However, the mechanism of Nef-induced activation of viral replication remains to be clearly understood. Using transient transfection assay, co-immunoprecipitation and pull-down analysis, we demonstrate in this report that the HIV-1 Nef protein physically interacts with Tat, the principal transactivating protein of HIV-1. Our observations with single cycle replication experiments further indicate that this interaction results not only in enhancement of Tat-induced HIV-1 long terminal repeat-mediated gene expression but also in virus production. PMID- 12885405 TI - Inhibition of FGF receptor signalling in Xenopus oocytes: differential effect of Grb7, Grb10 and Grb14. AB - The role of Grb7 adapters, Grb7, Grb10, and Grb14, was investigated in Xenopus oocytes expressing fibroblast growth factor receptors (FGFR). FGF-induced maturation of FGFR-expressing oocytes was blocked by previous injection of Grb7 or Grb14, but not Grb10. This effect correlated with Grb7/14 binding to the receptor, and inhibition of the Ras-dependent pathway. Interestingly, the phosphorylated insulin receptor interacting region (PIR) and Src 2 homology domains (SH2) of Grb7 and Grb14 were differently implicated in the inhibition of FGFR signalling. This study provided further evidence for specificity of the biological action of the Grb7 adapters on receptor tyrosine kinase signalling. PMID- 12885406 TI - Electrical activity in endocrine pituitary cells in situ: a support for a multiple-function coding. AB - The anterior pituitary is an endocrine gland that controls basic body functions. Pituitary cell functioning depends on membrane excitability, which induces cytosolic calcium rises. Here, we reported the first identification of small amplitude voltage fluctuations that controlled spike firing in endocrine cells recorded in situ. Three patterns of voltage fluctuations were distinguishable by their durations (1-100 s). These patterns could be ordered on top of each other, namely in response to secretagogues. Thus, pituitary endocrine cells express in situ a cell code in which small-amplitude voltage fluctuations lead to a multimodal arrangement of spike firing, which may finely tune calcium-dependent functions. PMID- 12885407 TI - Potassium uptake in the unicellular cyanobacterium Synechocystis sp. strain PCC 6803 mainly depends on a Ktr-like system encoded by slr1509 (ntpJ). AB - The molecular basis of potassium uptake in cyanobacteria has not been elucidated. However, genes known from other bacteria to encode potassium transporters can be identified in the genome of Synechocystis sp. strain PCC 6803. Mutants defective in kdpA and ntpJ were generated and characterized to address the role of the Kdp and KtrAB systems in this strain. KtrAB is crucial for K(+) uptake, as the DeltantpJ mutant shows slowed growth, slowed potassium uptake kinetics, and increased salt sensitivity. The DeltakdpA mutant has the same phenotype as the wild type even at limiting potassium, but a DeltakdpADeltantpJ double mutant is not viable, indicating a role of Kdp for potassium uptake when the Ktr system is not functioning. PMID- 12885408 TI - Pneumocystis carinii BCK1 functions in a mitogen-activated protein kinase cascade regulating fungal cell-wall assembly. AB - Pneumocystis pneumonia remains the most common AIDS-defining opportunistic infection in people with HIV. The process by which Pneumocystis carinii constructs its cell wall is not well known, although recent studies reveal that molecules such as beta-1-3-glucan synthetase (GSC1) and environmental pH responsive genes such as PHR1 are important for cell-wall integrity. In closely related fungi, a specific mitogen-activated protein kinase (MAPK) cascade regulates cell-wall assembly in response to elevated temperature. The upstream mitogen-activated protein kinase kinase kinase (MAPKKK, or MEKK), BCK1, is an essential component in this pathway for maintaining cell-wall integrity and preventing fungal cell lysis. We have identified a P. carinii MEKK gene and have expressed it in Saccharomyces cerevisiae to gain insights into its function. The P. carinii MEKK, PCBCK1, corrects the temperature-sensitive cell lysis defect of bck1Delta yeast. Further, at elevated temperature PCBCK1 restored the signaling defect in bck1Delta yeast to maintain expression of the temperature-inducible beta-1-3-glucan synthetase gene, FKS2. PCBCK1, as a functional kinase, is capable of autophosphorylation and substrate phosphorylation. Since glucan machinery is not present in mammals, a better understanding of this pathway in P. carinii might aid in the development of novel medications which interfere with the integrity of the Pneumocystis cell wall. PMID- 12885409 TI - Characterization of rapid and high-affinity uptake of L-serine in neurons and astrocytes in primary culture. AB - The non-essential amino acid L-serine was shown to be required to support the survival of rat cerebellar Purkinje neurons because of lack of the expression of the L-serine biosynthesis enzyme 3-phosphoglycerate dehydrogenase in them. In the present study, we investigated L-[(3)H]serine uptake in primary cultures of neurons and astrocytes from the rat telencephalon. In both neurons and astrocytes, L-[(3)H]serine uptake was dependent on temperature and Na(+) ions, and exhibited a single component of high-affinity uptake sites (K(m)=15.0 and 17.2 micro M for neurons and astrocytes, respectively). Kinetic analysis of L [(3)H]serine uptake also revealed that the uptake into neurons was faster than that into astrocytes. The selectivity of inhibition by amino acids of the L [(3)H]serine uptake resembled that of the system ASC transporters ASCT1 and ASCT2. Neutral amino acids L-alanine, L-serine, L-cysteine, and L-threonine strongly inhibited the uptake by both cell types. Furthermore, in astrocytes, but not in neurons, L-valine and L-proline also inhibited L-[(3)H]serine uptake. Neither alpha-methyl aminoisobutyric acid (a system A-specific substrate) nor 2 aminobicyclo(2,2,1)heptane-2-carboxylic acid (a system L-specific substrate) inhibited the uptake of L-[(3)H]serine in both neurons and astrocytes. Expression of ASCT transporters in both neurons and astrocytes was examined by use of reverse transcriptase polymerase chain reaction and immunoblot analysis. Whereas transcripts (mRNAs) of both ASCT1 and ASCT2 transporters were detected in astrocytes, only the mRNA of the former subtype was detected in neurons. Immunoblot analysis confirmed the presence of ASCT1 in both neurons and astrocytes. These findings indicate that neurons accumulate a high level of L serine by using a Na(+)-dependent, high-affinity transport system, operating predominantly through the ASCT1 transporter subtype. PMID- 12885410 TI - Esmolol is antiarrhythmic in doxorubicin-induced arrhythmia in cultured cardiomyocytes - determination by novel rapid cardiomyocyte assay. AB - Cardiac toxicity is an uncommon but potentially serious complication of cancer therapy, especially with anthracyclines. One of the most effective anticancer drugs is doxorubicin, but its value is limited by the risk of developing cardiomyopathy and ventricular arrhythmia. When applied to a network of periodically contracting cardiomyocytes in culture, doxorubicin induces rhythm disturbances. Using a novel rapid assay based on non-invasive ion-conductance microscopy we show that the beta-antagonist esmolol can restore rhythm in doxorubicin-treated cultures of cardiomyocytes. Moreover, esmolol pre-treatment can protect the culture from doxorubicin-induced arrhythmia. PMID- 12885411 TI - Cardiac hypertrophy in vivo is associated with increased expression of the ribosomal gene transcription factor UBF. AB - The ribosomal DNA transcription-specific factor, UBF, is a key target for the regulation of ribosomal RNA synthesis and hypertrophic growth of isolated neonatal cardiomyocytes. In this study, we have examined whether UBF expression is also an important determinant of cardiac growth rates in vivo. We show that rDNA transcription, rRNA synthesis and UBF expression in left ventricular myocytes isolated from mice 1-6 weeks following transverse aortic constriction were significantly increased (2.5-3.5-fold) compared to the levels in myocytes from the left ventricle of sham-operated mice. PMID- 12885412 TI - Prevention of passively transferred experimental autoimmune myasthenia gravis by an in vitro selected RNA aptamer. AB - Myasthenia gravis (MG) and its animal model, experimental autoimmune MG (EAMG), are mainly caused by autoantibodies directed against acetylcholine receptors (AChR) located in the postsynaptic muscle membrane. Previously, we isolated an RNA aptamer with 2'-fluoropyrimidines using in vitro selection techniques that acted as an effective decoy against both a rat monoclonal antibody called mAb198, which recognizes the main immunogenic region on the AChR, and a significant fraction of patient autoantibodies with MG. To investigate the therapeutic potential of the RNA, we tested the ability of the RNA aptamer to protect the receptors in vivo from mAb198. Clinical symptoms of EAMG in rats engendered by passive transfer of mAb198 were efficiently inhibited by a truncated RNA aptamer that was modified with polyethylene glycol, but not by control scrambled RNA. Moreover, the loss of AChR in the animals induced by the antibody was also significantly blocked with the modified RNA aptamer. These results suggested that RNA aptamers could be applied for antigen-specific treatment for autoimmune diseases including MG. PMID- 12885413 TI - New partners of acyl carrier protein detected in Escherichia coli by tandem affinity purification. AB - We report the first use of tandem affinity purification (TAP) in a prokaryote to purify native protein complexes, and demonstrate its reliability and power. We purified the acyl carrier protein (ACP) of Escherichia coli, a protein involved in a myriad of metabolic pathways. Besides the identification of several known partners of ACP, we rediscovered ACP/MukB and ACP/IscS interactions already detected but previously disregarded as due to contamination. Here, we demonstrate the specificity of these interactions and characterize them. This suggests that ACP is involved in additional previously unsuspected pathways. Furthermore, this study shows how the TAP method can be simply used in prokaryotes such as E. coli to identify new partners in protein-protein interactions under physiological conditions and thereby uncover novel protein functions. PMID- 12885414 TI - The ribosomal A site-bound sense and stop codons are similarly positioned towards the A1823-A1824 dinucleotide of the 18S ribosomal RNA. AB - Positioning of the mRNA codon towards the 18S ribosomal RNA in the A site of human 80S ribosomes has been studied applying short mRNA analogs containing either the stop codon UAA or the sense codon UCA with a perfluoroaryl azide group at the uridine residue. Bound to the ribosomal A site, a modified codon crosslinks exclusively to the 40S subunits under mild UV irradiation. This result is inconsistent with the hypothesis [Ivanov et al. (2001) RNA 7, 1683-1692] which requires direct contact between the large rRNA and the stop codon of the mRNA as recognition step at translation termination. Both sense and stop codons crosslink to the same A1823/A1824 invariant dinucleotide in helix 44 of 18S rRNA. The data point to the resemblance between the ternary complexes formed at elongation (sense codon.aminoacyl-tRNA.AA dinucleotide of 18S rRNA) and termination (stop codon.eRF1.AA dinucleotide of 18S rRNA) steps of protein synthesis and support the view that eRF1 may be considered as a functional mimic of aminoacyl-tRNA. PMID- 12885415 TI - MyD88S, a splice variant of MyD88, differentially modulates NF-kappaB- and AP-1 dependent gene expression. AB - MyD88 is an adapter protein that is involved in Toll-like receptor (TLR)- and interleukin-1 receptor (IL-1R)-induced activation of nuclear factor-kappaB (NF kappaB) and c-Jun N-terminal kinase (JNK). By directly binding IL-1R-associated kinase (IRAK)-1 and IRAK-4, MyD88 serves as a bridging protein, enabling IRAK-4 induced IRAK-1 phosphorylation. We previously identified a lipopolysaccharide inducible splice variant of MyD88, MyD88(S), which specifically prevents the recruitment of IRAK-4 into the IL-1R complex and thus inhibits IRAK-4-mediated IRAK-1 phosphorylation. MyD88(S) is not able to activate NF-kappaB, and in contrast functions as a dominant negative inhibitor of TLR/IL-1R-induced NF kappaB activation. Unexpectedly, we here demonstrate that MyD88(S) still allows JNK phosphorylation and activator protein (AP)-1-dependent reporter gene induction upon overexpression in HEK293T cells. These observations indicate that NF-kappaB and JNK activation pathways can already diverge at the level of MyD88. Moreover, the regulated expression of a MyD88 splice variant which specifically interferes with NF-kappaB- but not AP-1-dependent gene expression implies an important role for alternative splicing in the fine-tuning of TLR/IL-1R responses. PMID- 12885416 TI - Differential binding of Sin3 interacting repressor domains to the PAH2 domain of Sin3A. AB - The Sin3 interacting domain (SID), originally described in the Mad family of repressors, is a novel transcriptional repressor domain that binds the PAH2 domain of corepressors Sin3A and Sin3B with high affinities. The conserved SID like domains are reportedly present in five KLF proteins. However, the KLF SIDs and the Mad SIDs can be classified into two subtypes according to sequence similarity. Here, we report the finding from computational and experimental studies that the two subtypes of SID domains bind differentially to Sin3A. This finding offers insights into a mechanism of cell growth regulation by interactions of different subtypes of SID-containing repressor proteins with Sin3. It also provides the structural basis for developing selective modulators of Sin3. PMID- 12885417 TI - Gene silencing in mammalian cells by PCR-based short hairpin RNA. AB - RNA interference (RNAi) provides a powerful tool to silence genes in a sequence specific manner in a variety of systems. However, not all sequences are effective in the RNAi-mediated gene silencing. In this study, we developed a polymerase chain reaction (PCR)-based RNAi strategy for a quick screening of small interfering RNA (siRNA) efficiency. This method utilized a two-step PCR to generate a chimeric DNA template containing the U6 promoter or cytomegalovirus promoter and short hairpin DNA. We demonstrated that the transfection of the PCR products into mammalian cells resulted in specific depressions of exogenous (luciferase, green fluorescent protein and beta-galactosidase) and endogenous (annexin II) gene expressions. This PCR strategy provides a rapid, easy and cheap approach for testing candidates siRNA sequences and is an attractive alternative to subcloning. PMID- 12885418 TI - The membrane bound N-terminal domain of human adenosine diphosphate ribosylation factor-1 (ARF1). AB - The small G protein adenosine diphosphate ribosylation factor-1 (ARF1) is activated by cell membrane binding of a self-folding N-terminal domain. We present a model of the human ARF1 N-terminal peptide in planar lipid bilayers, determined from neutron lamellar diffraction and circular dichroism data with molecular modelling. This amphipathic domain lies at a shallow membrane depth, ideal for regulation of the ARF1 bio-timer by rapid, reversible membrane binding. The helical region does not elongate upon membrane binding, leaving the connecting flexible linker region's length unchanged. PMID- 12885419 TI - Macrophage inflammatory protein-related protein-2, a novel CC chemokine, can regulate preadipocyte migration and adipocyte differentiation. AB - Adipocytes not only store energy, but also secrete biologically active molecules called adipocytokines, which play a pivotal role in adipocyte-related pathological processes such as diabetes and cardiovascular disease. Recent studies have shown that preadipocyte/adipocyte expresses chemokines (e.g. monocyte chemoattractant protein-1, macrophage inflammatory protein-1 alpha) which alter adipocyte function, indicating the involvement of chemokines in adipocyte-related pathologies. The current study investigated the potential of macrophage inflammatory protein-related protein-2 (MRP-2), a novel CC chemokine, to modulate preadipocyte trafficking and adipocyte differentiation. MRP-2 and its receptors were highly expressed in preadipocytes and differentiated adipocytes as well as in the mouse fat pad. Chemotaxis assays revealed that MRP-2 was a specific chemotactic regulator in preadipocyte migration. The levels of MRP-2 expression in adipose tissue were enhanced in obese mice compared to lean mice. MRP-2 secretion by preadipocytes was suppressed during differentiation. MRP-2 suppressed the expression of adipocyte differentiation markers such as adipocyte fatty acid-binding protein and glycerol-3 phosphate dehydrogenase. Taken together, our data suggest that MRP-2 plays a role in the regulation of preadipocyte migration and adipocyte differentiation during adipose tissue development. MRP-2 may be another adipocytokine, which can be involved in the adipocyte-related pathological process. PMID- 12885420 TI - Mucosal activity patterns as a basis for olfactory discrimination: comparing behavior and optical recordings. AB - In over half a century numerous studies have demonstrated that different odorants produce individually different spatial patterns of neural receptor activity on the olfactory mucosa. However, the thought that these differential activity patterns could be the neural code underlying olfactory perception has not been tested directly. In the present study using operant techniques, rats were trained to differentially identify five odors from a homologous series of iso-intensive straight-chain aldehydes that differed serially by only one carbon atom, viz. hexaldehyde to decaldehyde. The rats identified each of the five odorants with greater than 90% correct identification. The degree of perceptual similarity between any pair of the five odorants was determined. Using multidimensional scaling techniques (MDS) the similarity measures yielded a two-dimensional perceptual odorant space. Optical techniques were used to record the olfactory mucosal activity patterns in response to these same five iso-intensive aldehydes. The mucosal activity elicited by each odorant revealed individually distinct band like patterns that varied both within and across these bands. More importantly, the relative differential responsivity of the bands was related to chain length. An MDS analysis of the dissimilarity measure between all possible pairs of odorant induced activity patterns yielded a two-dimensional neurophysiologic odorant space. Further analysis indicated that the neurophysiologic and psychophysically determined odorant spaces were highly correlated (F(1,39)=23.9, P=nil). These results give additional credence to the concept that the odorant induced mucosal activity patterns may serve as the substrate for the perception of odorant quality. PMID- 12885421 TI - Role of central NMDA versus non-NMDA receptor in spinal withdrawal reflex in spinal anesthetized rats under normal and hyperexcitable conditions. AB - The present study aimed to investigate the role of central N-methyl-D-aspartate (NMDA) and non-NMDA receptors in the spinal withdrawal reflex assessed by recording single motor unit (SMU) electromyogram (EMG) response to peripheral mechanical (pressure, pinch) stimuli and repeated electrical stimuli at 3 and 20 Hz. During normal conditions, intrathecal administration of MK-801 and CNQX apparently depressed mechanically and electrically (3 Hz) evoked EMG responses in a dose-dependent manner (10, 20 and 40 nmol in 10 microl). In contrast, the after discharges to 20 Hz electrical stimuli were suppressed only by CNQX treatment, not by MK-801 treatment. This indicates that the central mechanisms underlying the different frequencies of electrically evoked withdrawal reflex may be different. During peripheral bee venom (BV, 0.2 mg/50 microl) induced inflammation and central sensitization, the enhanced SMU EMG responses including after-discharges to pinch stimuli and 3 Hz electrical stimuli were depressed significantly by treatments with both MK-801 and CNQX. However, the enhanced SMU activities to innocuous pressure stimuli were depressed only by treatment with CNQX. Likewise, enhanced long lasting after-discharges elicited by 20 Hz electrical stimuli were also only depressed by CNQX, indicating that different central mechanisms are involved in the persistent hyperexcitability during BV induced inflammation. The data suggest that both central NMDA and non-NMDA receptors play important roles in the transmission of nociceptive information under normal conditions. In BV-induced inflammation, however, central non-NMDA receptors, but not NMDA receptors, play a pivotal role in the generation of persistent hyperexcitability to mechanical and electrical stimuli at different frequencies (3 Hz, 20 Hz). PMID- 12885422 TI - Brain opioid-receptors are involved in mediating peripheral electric stimulation induced inhibition of morphine conditioned place preference in rats. AB - Conditioned place preference (CPP) paradigm has been suggested as one of the animal models for drug craving. The present study was performed to examine the effect of 100 Hz peripheral electric stimulation (PES) on the expression of morphine-induced CPP. Rats were trained with morphine for 4 days to establish the CPP paradigm in a three-chamber "unbiased" apparatus. Morphine-induced CPP was maintained up to 4 weeks when tests were given once a week. PES of 100 Hz administered 30 min a day for 3 days significantly attenuated morphine-induced CPP (P<0.01). I.c.v. injection of the delta-opioid receptor antagonist naltrindole (NTI) or the kappa-antagonist norbinaltorphimine (nor-BNI) but not the mu-antagonist cyclic D-Phe-Cys-Tyr-D-Trp-Arg-Thr-Pen-Thr-NH(2) (CTAP), completely blocked the inhibitory effect of 100 Hz PES on the expression of morphine-induced CPP (P<0.05-0.01). These results indicate that the anti-craving effects induced by repeated PES of 100 Hz is mediated by the activation of supra segmental delta- and kappa-opioid receptors in the central nervous system. PMID- 12885423 TI - Serum-derived factors weaken the barrier properties of cultured porcine brain capillary endothelial cells in vitro. AB - Cultured cerebral capillary endothelial cells are often used as a functional in vitro model of the blood-brain barrier (BBB) to determine drug uptake or to study barrier properties. Usually serum is supplemented to these cultures for cell proliferation. Here, we demonstrate the effect of serum and the serum-derived factors lysophosphatidic acid (LPA) and vascular endothelial growth factor (VEGF) on the barrier properties of cultured porcine brain capillary endothelial cells (PBCEC). Serum prevents tight junction formation of confluent PBCEC monolayers and moreover, opens already established tight junctions shown by decreasing transendothelial electrical resistances (TER). These effects are highly polarised with serum almost exclusively acting from the basolateral side of the cell culture. Immunocytochemistry of PBCEC revealed a delocalisation of the cell border lining tight junction proteins ZO-1, occludin and claudin-5 when serum was added. A serum fraction of 67 kDa was isolated by size-exclusion chromatography, identified as albumin and found to cause a serum-like decrease of the TER. However, fatty acid-free serum albumin does not develop this barrier weakening effect, indicating that small protein-bound factors might be responsible. For instance, serum-bound LPA demonstrated a TER-decreasing effect as well, but in contrast to serum mainly when added to the apical side of PBCEC. Addition of VEGF caused a serum-like decrease of the TER with the same polar effect; however, VEGF will be denatured by heat and could thus not be the heat-sensitive factor. Thus, we hypothesise that serum contains a variety of factors which weaken the tightness of a PBCEC monolayer from the apical side as expected but also from the basolateral side. Although the structure of the 67 kDa factor could not be analysed, this finding is of importance for in vitro models not only of the blood brain barrier mostly using serum-containing media. PMID- 12885424 TI - Subtypes of substance P receptor immunoreactive interneurons in the rat basolateral amygdala. AB - Local injections of the neurotoxin SP-saporin into the basolateral amygdala (BLA) are reported to specifically lesion substance P receptor immunoreactive (SPR-IR) interneurons, and to reduce anxiety related behavior. Hence, this technique might provide a means to study how defined interneuron populations regulate neuronal activity in the BLA. However, what interneuron subgroups in the BLA might be targeted by SP-saporin lesions has not been established. This study has used dual labeling immunofluorescence in the rat BLA to examine SPR-IR neurons for their colocalization with the calcium-binding proteins; calbindin-D28k (CB), parvalbumin (PV), and calretinin (CR); and the neuropeptides somatostatin (SOM) and neuropeptide Y (NPY). We found that all NPY-IR neurons and 45% of SOM-IR interneurons expressed SPR-IR, and that 50% and 51% of the SPR-IR interneuron population expressed NPY- and SOM-IR, respectively. Previous studies have reported that approximately a third of SOM-IR interneurons also express NPY, which suggests a large degree of overlap between the NPY, SOM and SPR expressing neurons in the BLA. We also found that the majority of SPR-IR cells were CB-IR (62%), but that these interneurons represented only 2.8% of the total CB-IR population. Moreover, SPR-IR interneurons did not express either PV-or CR- IR. Hence, SP-saporin lesions would ablate all interneurons in the BLA that contain NPY, but leave the majority of the CB-IR cells intact, and have no effect on the CR- and PV-IR populations. Consequently, these results support the use of SP saporin lesions as a useful technique to study the role of NPY-IR interneurons in the BLA. PMID- 12885425 TI - Membrane and synaptic effects of corticotropin-releasing factor on periaqueductal gray neurons of the rat. AB - Corticotropin-releasing factor (CRF) has been identified as a major component of the hypothalamic-pituitary-adrenal (HPA) axis. By stimulating the release of adrenocorticotropin hormone (ACTH), CRF acts as a key mediator of the stress response. However, CRF receptors and neuronal elements are present in many extrahypothalamic regions of the brain. A region that contains both CRF-ergic neurons and CRF receptors is the midbrain periaqueductal gray (PAG). The physiological effects of CRF in the PAG are unknown. In this study, an in vitro preparation, extracellular and intracellular patch-clamp recordings, were used to examine the effects of CRF, applied through an injecting electrode, on PAG neurons. Recordings were made from 147 neurons in the PAG. CRF injecting electrode concentrations of 0.05 and 1 microM were tested. At the higher concentration, CRF had a predominant excitatory effect on the neurons, and at the lower concentration, CRF produced no significant effect on the neurons. The excitatory effect was dose dependent and was often associated with a depolarization in membrane potential in intracellular recordings. Application of the CRF antagonist, alpha-helical CRF, blocked this excitatory effect. It is concluded that CRF has a predominant excitatory effect on PAG neurons. It is also concluded that CRF is not acting presynaptically. This excitatory effect of CRF on PAG neurons may lead to activation of a descending analgesic pathway. PMID- 12885426 TI - Cerebral angiogenesis and expression of VEGF after subarachnoid hemorrhage (SAH) in rats. AB - Subarachnoid hemorrhage (SAH) leads to the development of vasospasm in which endothelin-1 plays a very important role. The effect of its vasoconstricting action is hypoxia of the nervous tissue, which stimulates the release of growth factors. Vascular endothelial growth factor (VEGF) released in excessive amounts from hypoxically altered cerebrovascular endothelial cells is the most potent angiogenic factor and may enhance angiogenesis after SAH. If endothelin-1 is mainly responsible for vasospasm after SAH, it is possible that early administration of endothelin converting enzyme inhibitor or endothelin receptor antagonist may protect neurons against. The aim of the study was to establish whether prolonged vasospasm and endothelial cell hypoxia stimulate VEGF expression and, in consequence, promote angiogenesis in the central nervous system after subarachnoid hemorrhage. Investigations were also performed to determine whether the administration of phosphoramidon, an endothelin-converting enzyme (ECE) inhibitor, and BQ-123, an endothelin receptor ET(A) antagonist, suppresses angiogenesis and VEGF expression. Experiments were carried out in male Wistar rats injected with phosphoramidon or BQ-123 into the cisterna magna following the induction of subarachnoid hemorrhage. The brains were removed 48 h after the hemorrhage for histopathological and immunohistochemical examinations of VEGF expression and angiogenesis in the cerebral hemispheres, brainstem, and cerebellum. Statistical analysis was performed using nonparametric Wilcoxon test (P<0.05). The results obtained have shown for the first time a close correlation between endothelial hypoxia after SAH in cerebral microvessels and enhanced angiogenesis. There is also an increase in VEGF expression in cerebral vessels and neurons within the cerebral hemispheres, brainstem, and cerebellum. The administration of phosphoramidon or BQ-123 has been found to inhibit angiogenesis. Angiogenesis in the chronic phase of SAH-induced vasospasm is the result of prolonged narrowing of vessels due to excessive secretion of endothelin by damaged endothelial cells. Present results obtained indicate that it is possible to reduce or prevent the late effects of SAH, i.e., neuronal hypoxia and cerebral edema, through the inhibition of endothelin-1 induced vasospasm. PMID- 12885427 TI - Three kinds of current in response to substance P in bullfrog DRG neurons. AB - In response to SP applied externally, neurons freshly isolated from bullfrog dorsal root ganglion (DRG) showed three kinds of current (I(SP)), i.e. slow, fast and moderately activating I(SP)s. All the three kinds of I(SP) were inward currents, and were completely blocked by either peptide antagonist of SP receptor spantide or non-peptide antagonist of SP receptor WIN51708. The slow activating I(SP) showed slow kinetic features. Replacement of NaCl in external solution by NMDG had no effect on this kind of I(SP), while Ba(2+) abolished it almost completely, thus the ionic mechanism underlying slow activating I(SP) was deduced to be the closure of K(+) channels. The fast activating I(SP) in bullfrog DRG neurons, just as in rat DRG neurons, was proved to be caused by the opening of Na(+) preferring non-selective cation channel, for it was abolished almost completely by replacement of NaCl in external solution with equimolar NMDG. The moderately activating I(SP) was similar to the fast activating I(SP) in current configuration, however, its kinetic characteristics lay between those of fast and slow activating I(SP)s. Either NMDG or Ba(2+) suppressed this kind of I(SP) partially. Therefore the moderately activating I(SP) might be mediated by non selective cation channel. We used repatch technique to explore the intracellular mechanism underlying the three kinds of I(SP) and found that the three kinds of I(SP) were caused by the activity of either G-protein coupled channel (slow activating I(SP)) or directly opened channel (fast activating I(SP)) or both (moderately activating I(SP) ). PMID- 12885428 TI - A possible novel mechanism underlying temperature-dependent uptake of [3H]spermidine in nuclear fractions of murine brain. AB - [3H]Spermidine (SPD) was accumulated in subcellular fractions enriched of the nucleus in a temperature-dependent manner with a saturable profile in murine brain. The accumulation reached a plateau within 30 min at 2 degrees C and 30 degrees C, while excess unlabeled SPD significantly inhibited the accumulation at 2 degrees C without markedly affecting that at 30 degrees C when added after equilibrium. The temperature-dependent accumulation of [3H]SPD was significantly inhibited by the triamine SPD and the tetraamine spermine, but not by the diamine putrescine. Phospholipases were invariably effective in significantly inhibiting the accumulation at 30 degrees C in a concentration-dependent manner. Amongst different discrete murine central structures examined, the temperature-dependent [3H]SPD accumulation was highest in neocortex with progressively lower activities in striatum, hypothalamus, spinal cord, medulla-pons, hippocampus, midbrain and cerebellum. These results suggest the possible presence of a hitherto unidentified nuclear transport system for particular polyamines in murine brain. PMID- 12885429 TI - In vitro pharmacological characterization of (+/-)-4-[2-(1-methyl-2 pyrrolidinyl)ethyl]thio]phenol hydrochloride (SIB-1553A), a nicotinic acetylcholine receptor ligand. AB - SIB-1553A ((+/-)-4-[2-(1-methyl-2-pyrrolidinyl)ethyl]thio]phenol HCl) is a neuronal nicotinic acetylcholine receptor (nAChR) ligand which displaced the binding of [3H]nicotine (NIC) to the rat brain nAChRs with an IC(50) value of 110 nM with no appreciable affinity to the alpha7 nAhRs. SIB-1553A showed modest affinity for histaminergic (H3) and serotonergic (5-HT1 and 5-HT2) receptors, and sigma binding sites. In calcium flux assays, SIB-1553A (0.1-5 microM), in contrast to nicotine, showed a greater selectivity for beta4-subunit containing recombinant hnAChRs (alpha2beta4, alpha3beta4 and alpha4beta4) vs. beta2-subunit containing nAChRs (alpha4beta2 and alpha3beta2) both in terms of efficacy and potency. While NIC (10-30 microM) and epibatidine (0.01-0.1 microM) fully activated human muscle-type AChRs expressed by RD cell line, SIB-1553A was virtually ineffective for up to >100 microM and elicited less than 10% of the response due to suberyldicholine. SIB-1553A (< or =30 microM) evoked [3H]DA release from striatum, olfactory tubercles and prefrontal cortex (PFC), and [3H]NE release from hippocampus and PFC, and this evoked release was sensitive to mecamylamine (MEC). SIB-1553A-evoked neurotransmitter release exhibited region- and transmitter-specific antagonism by dehydro-beta-erythroidine (DHbetaE). SIB 1553A was less efficacious than NIC at evoking [3H]NE from the rat hippocampus and antagonized NIC response upon co-application implying partial agonist properties. SIB-1553A did not evoke basal [3H]ACh release from the rat striatum or hippocampus, but attenuated NMDA-evoked [3H]ACh release from the rat striatum. SIB-1553A did not inhibit rat brain cholinesterase for up to 1 mM. Multiple receptor affinities and release of several neurotransmitters may underlie the cognitive-enhancing effects of SIB-1553A documented in rodent and primate models. PMID- 12885430 TI - Effects of chronic dizocilpine on acute pain and on mRNA expression of neuropeptides and the dopamine and glutamate receptors. AB - The mesocorticolimbic circuitry has been implicated in the pathophysiology of several neuropsychiatric syndromes like chronic pain and addiction. The aim of this study was to evaluate the effects of dizocilpine (MK-801), a non-competitive N-methyl-D-aspartate (NMDA) receptor antagonist, on sensorimotor behaviors and the consequent changes in the dopamine, glutamate, and opiate systems in rats. Five groups of rats were subjected to acute tests for nociception (hot plate and paw pressure) before and after MK-801 (0.05, 0.1, 0.2 and 0.4 mg/kg, i.p.) or saline. Another two groups received daily i.p. saline or MK-801 (0.4 mg/kg) for 15 days. The nociceptive tests were performed on days 1, 7, and 14. On day 15 the rats received the last injection and were immediately sacrificed. We measured the mRNA expression, by in situ hybridization (ISH), of various dopamine and glutamate receptors, and enkephalin (Enk), dynorphin (Dyn), and substance P (SP) in the striatum, nucleus accumbens (NAC), piriform and cingulate cortex. Acute MK 801, dose-dependently, resulted in hyperalgesia. The chronic effects of 0.4 mg/kg MK-801 showed an extinction of the acute hyperalgesic effects especially with the hot plate test. The ISH studies revealed a decrease in mRNA expression of Enk and SP in the striatum and NAC. Our results indicate that the reversal of acute MK 801-induced hyperalgesia, with repeated exposure to systemic MK-801, is not directly related to changes in dopamine and glutamate receptors and might involve alteration of the striatal neuropeptide system. PMID- 12885431 TI - Nestin expression after experimental intracerebral hemorrhage. AB - The current study examines nestin expression after intracerebral hemorrhage (ICH), the role of different blood components in nestin upregulation, and the possibility that low doses of thrombin that induce tolerance to brain injury (thrombin preconditioning) might also induce nestin expression. Adult male Sprague-Dawley rats received an intracaudate injection of either whole blood, thrombin (1 or 5 U) or red blood cells (RBCs). Animals were sacrificed for single and double labeling immunohistochemistry to identify which cells express nestin, and for Western blotting to quantify nestin expression. By immunohistochemistry, nestin immunoreactivity was present in large numbers of astrocytes, surrounding the hematoma from day 3 to 1 week after ICH. After 2 weeks, nestin immunoreactivity was co-localized with a neuronal marker (neuronal specific enolase). By Western blot analysis, nestin was strongly expressed at day 3 (P<0.01) and 1 week (P<0.01), and expression persisted for at least 1 month (P<0.05). Intracerebral injection of thrombin or lysed RBCs resulted in a marked increase in nestin expression. Interestingly, injection of a low dose of thrombin that induces brain tolerance also upregulated nestin. The ICH-induced nestin expression in astrocytes may reflect an early response of these cells to injury, while the delayed expression in neurons might be a part of the adaptative response to injury perhaps leading to recovery of function. Nestin induction by a low dose of thrombin suggests that specific receptor-mediated pathways are involved in inducing nestin expression and that nestin may play a role in thrombin preconditioning. PMID- 12885432 TI - Long-lasting CNS effects of a short-term chemical knockout of ornithine decarboxylase during development: nicotinic cholinergic receptor upregulation and subtle macromolecular changes in adulthood. AB - Ornithine decarboxylase (ODC) and the polyamines play an essential role in brain cell replication and differentiation and polyamines also regulate the function of nicotinic acetylcholine receptors (nAChRs). We administered alpha difluoromethylornithine (DFMO), an irreversible inhibitor of ODC, to neonatal rats on postnatal days 5-12, during the mitotic peak of the cerebellum, a treatment regimen that achieves a chemical knockout of ODC activity and polyamine depletion limited to the treatment period. Although growth inhibition and gross dysmorphology were limited to the cerebellum, both alpha7 and alpha4beta2 nAChRs were upregulated in adulthood in the frontal cortex, hippocampus and thalamus, with the largest effect in the latter region, primarily in females. Receptor upregulation was accompanied by abnormalities in macromolecular indices of cell packing density and cell membrane surface area, but the generalized cellular alterations did not share the regional or sex selectivity shown by the effects on nAChRs. Elevated DNA concentration was most notable in the hippocampus and was associated with augmented levels of glial fibrillary acidic protein, thus implying gliosis as the cause of the increased number of cells. DFMO's effects on both nAChR expression and cellular biomarkers resembled those of developmental exposure to nicotine. Accordingly, some of the effects may represent a specific alteration in nAChR signaling evoked by polyamine depletion during a critical developmental window. Alterations in polyamine gating of cholinergic synaptic signaling may thus contribute to the adverse neurobehavioral effects of numerous neuroteratogens that directly or indirectly disrupt the ODC/polyamine pathway. PMID- 12885433 TI - Reciprocal serotonergic connections between the hamster median and dorsal raphe nuclei. AB - The median (MnR), but not the dorsal (DR) raphe, sends a serotonergic projection to the suprachiasmatic (SCN) nucleus. Stimulation of either nucleus by electrode or serotonin agonist yields equivalent effects on circadian rhythmicity. This and other evidence suggests the existence of a functional serotonergic pathway from the DR to the MnR that may participate in circadian rhythm regulation. The present investigation was designed to identify such a connection. Tract tracer studies revealed cells in the DR that project to the MnR, as well as cells in the MnR that project to the DR. Double label immunofluorescence methods demonstrated that some of the cells projecting from either nucleus to the other contain serotonin immunoreactivity. The results support the existence of a reciprocal pathway between the DR and MnR that is at least partially serotonergic. PMID- 12885434 TI - Tumor necrosis factor alpha increases cytosolic calcium responses to AMPA and KCl in primary cultures of rat hippocampal neurons. AB - Acute behavioral effects of tumor necrosis factor alpha (TNFalpha) have been previously reported, however the cellular basis for these actions are unknown. To address this issue we examined the effects of TNFalpha on AMPA- and depolarization-induced changes in cytosolic Ca(2+) in cultured hippocampal neurons. Single cell Ca(2+) levels were determined with the fluorescent calcium indicator fura-2. TNFalpha caused an up-regulation of AMPA (10 microM)- and depolarization (55 mM KCl)-induced Ca(2+) responses. This effect occurred within a window of concentrations (1 and 10 ng/ml but not 0.1 or 100 ng/ml) and times (3 and 6 h but not 1 and 24 h). The effect was dependent upon protein synthesis (blocked by cycloheximide) and was prevented by the soluble TNF receptor and by a soluble TNF receptor fragment. Treatment with the soluble TNF receptor fragment also caused a decrease in the basal response. The TNFalpha treatment protocols did not appear to produce any toxicity to the neurons. Results are consistent with the hypothesis that TNFalpha regulates proteins known to be involved in neuronal communication (AMPA receptors) and cell regulation (voltage-dependent calcium channels) in a relatively rapid period of time (a few hours). These actions may be related to the behavioral effects produced by TNFalpha that occur within this time frame. PMID- 12885435 TI - Effects of central hypocretin-1 administration on hemodynamic responses in young adult and middle-aged rats. AB - The prevalence of hypertension in middle age correlates with impaired autonomic regulation and as norepinephrinergic neurons decline with increasing age, and this reduction may contribute to this impairment. Central hypocretin-activated norepinephrinergic neurons contribute to sympathetic regulation. In the present study we compared sympathoadrenal effects of intracerebroventricular (i.c.v.) hypocretin-1(5 nmol) between young-adult (12-14 weeks) and middle-aged (12-14 months) rats. Arterial blood pressure, heart rate and plasma catecholamines were assessed under pentobarbital anesthesia. In addition, we compared hypocretin-1 and K(+)-evoked norepinephrine release from the cerebrocortical slices prepared from young-adult and middle-aged rats. We also examined whether the novel hypocretin receptor-1 antagonist (SB-334867) could reverse these hypocretin-1 effects both in vivo and in vitro. I.c.v. hypocretin-1 significantly increased blood pressure by some 7%, heart rate by 9% and plasma norepinephrine concentrations by 100% in young-adult rats. In middle-aged rats these parameters did not change. Plasma epinephrine did not increase in either group. There was a significant correlation between changes in mean arterial pressure and plasma norepinephrine. Similarly, hypocretin-1 evoked norepinephrine release from cerebrocortical slices prepared from young-adult rats was significantly higher than that of middle-aged rats whilst K(+)-evoked release did not differ between the groups. SB-334867 significantly attenuated hypocretin-1-increased blood pressure and both in vivo and in vitro norepinephrine release. The present data suggest that hypocretinergic neurons may contribute to the regulation of central but not adrenal sympathetic activity. Moreover, sympathetic regulation by hypocretinergic neurones may disappear in middle-age in rats. PMID- 12885436 TI - Central 5-HT2B/2C and 5-HT3 receptor stimulation decreases salt intake in sodium depleted rats. AB - In the present study, we investigated the participation of central 5-HT(2B/2C) and 5-HT(3) receptors in the salt intake induced by sodium depletion in Wistar male rats. Sodium depletion was produced by the administration of furosemide associated with a low salt diet. Third ventricle injections of mCPP, a 5 HT(2B/2C) agonist, at doses of 80, 160 and 240 nmol, promoted a dose-dependent reduction in salt intake in sodium-depleted rats. The inhibitory effect produced by central administration of mCPP was abolished by the central pretreatment with SDZ SER 082, a 5-HT(2B/2C) antagonist. Similar results were obtained with third ventricle injections of m-CPBG (80, 160 and 240 nmol), a selective 5-HT(3) agonist that also induced a dose-related decrease in salt intake in sodium depleted rats. The central pretreatment with LY-278,584, a selective 5-HT(3) receptor antagonist, was able to impair the salt intake inhibition elicited by third ventricle injections of m-CPBG. Central administration of each one of the antagonists alone or a combination of both antagonists together did not significantly change salt intake after sodium depletion. On the other hand, the central administration of both mCPP and m-CPBG, in the highest dose used to test their effect on salt intake (240 nmol), was unable to modify blood pressure in sodium-depleted rats. It is concluded that: (1) pharmacological activation of central 5-HT(2B/2C) and 5-HT(3) receptors diminishes salt intake during sodium depletion, (2) an inhibitory endogenous drive exerted by central 5-HT(2B/2C) and 5-HT(3) receptors does not seem to exist and (3) the reduction in salt intake generated by the pharmacological activation of these central receptors is not produced by an acute hypertensive response. PMID- 12885437 TI - Basolateral amygdala efferents to the ventral subiculum preferentially innervate pyramidal cell dendritic spines. AB - The basolateral amygdala and the ventral subiculum of the hippocampal formation are two of the major limbic-related regions within the brain, both of which project heavily to the nucleus accumbens. The nucleus accumbens is regarded as the limbic-motor interface, in view of these limbic afferent and its somatomotor and autonomic efferent connections. These afferent inputs have been suggested to converge monosynaptically on cells within the accumbens and are hypothesised to play a role in functions such as affective motivational behaviour. Convergence between inputs from the basolateral amygdala and the hippocampus at the level of the accumbens can be demonstrated with electrophysiological recording methods, but these do not conclusively preclude polysynaptic mechanisms. In fact there is a robust reciprocal projection between the basolateral amygdala and the hippocampus, synaptic details of which have not been fully investigated. We examined the synaptic input from the basolateral amygdala to the projection neurons of the subiculum, the spiny pyramidal neurons. We labelled the afferents from basolateral amygdala with a small injection of biotinylated dextran amine, and revealed the anterogradely labeled fibers within the subiculum. The labeled basolateral amygdala fibers were studied with electron microscopy to identify their postsynaptic target structures. With this technique we have demonstrated anatomically that the basolateral amygdala preferentially innervates spiny subiculum neurons, presumed pyramidal projection neurons, although some dendrites and possibly local circuit neurons may be targeted. PMID- 12885438 TI - Polyamines in a genetic animal model of paroxysmal dyskinesia. AB - Previous studies suggested that glutamatergic overactivity contributes to the manifestation of dystonia in the dt(sz) mutant hamster, a model of idiopathic paroxysmal dyskinesia in which dystonic episodes occur in response to mild stress. Therefore, the role of polyamines, known as positive modulators of NMDA receptors, was examined in the present study. The levels of polyamines (putrescine, spermidine, spermine) were determined in forebrain, cerebellum and brainstem in dt(sz) hamsters at an age of most marked expression of dystonia (32 days) and in age-matched non-dystonic control hamsters. Spermine was found to be significantly increased in the forebrain (35%) of dystonic animals, while spermidine was unaltered in dystonic brains and only a moderate increase in putrescine (12%) was detected in the cerebellum of dt(sz) mutants. In view of enhanced spermine levels, the effect of the putative polyamine receptor antagonist ifenprodil on the severity of dystonia was examined in dystonic hamsters. Ifenprodil (5-40 mg/kg i.p.) failed to exert a beneficial effect, but even aggravated dystonia in the dt(sz) mutant at higher doses. These data together with previous pharmacological findings in mutant hamsters do not completely exclude a pathophysiological role of enhanced polyamine levels but suggest that overstimulation of NMDA receptors which contain NR2B subunits by enhanced spermine levels is not involved in the dystonic syndrome. PMID- 12885439 TI - Adenosine triphosphate inhibits cytokine release from lipopolysaccharide activated microglia via P2y receptors. AB - Microglial proliferation and activation have been reported to occur after several central nervous system injuries. In this study, we tested the effects of adenosine triphosphate (ATP) on cultured microglia obtained from the spinal cord of rat embryos. The amounts of tumor necrosis factor alpha (TNF-alpha), interleukin 1beta and interleukin 6 released from the microglia, which were stimulated by lipopolysaccharide (LPS; 100 ng/ml), were inhibited by the simultaneous addition of ATP in a dose dependent manner (10-300 microM). We examined the effect of several endogenous purines (ATP, ADP, CTP, UDP, UTP) and P(2)y receptor agonists (ADPbetaS and 2-methylthio-ATP) on LPS-induced TNF-alpha release. The rank order of inhibitory potency of endogenous purines on TNF-alpha release was: ATP>ADP>>UTP>UDP>CTP. The latter three were much less potent than the former two. The addition of 10 microM 2-methylthio-ATP showed a potency similar to 100 microM ATP. The addition of ADPbetaS, however, showed less effect. These endogenous purines and selective ATP receptor agonists showed a similar inhibitory effect in their rank order on LPS-induced interleukin 6 release. We demonstrate that ATP inhibits cytokine release from LPS-activated microglia via metabotropic receptors. The application of P(2)y receptor agonists might be considered as a pharmacological treatment of several pathological conditions of the spinal cord, including toxic immunoreactions. PMID- 12885440 TI - Prevention of kainic acid seizures-induced changes in levels of nitric oxide and high-energy phosphates by 7-nitroindazole in rat brain regions. AB - Previous studies using the spin trapping agent N-tert-butyl-alpha-phenylnitrone (PBN) and the antioxidant vitamin E established the involvement of free radicals in kainic acid (KA)-induced neurotoxicity. In the present study, we examined the effects of the neuronal nitric oxide synthase (nNOS) inhibitor 7-nitroindazole (7 NI) to establish a possible role of nitric oxide (NO) in the neurotoxicity caused by KA-induced status epilepticus (SE). A single injection of KA (15 mg/kg, s.c.) induced seizures within 40-45 min, progressing to full seizure activity lasting about 3 h. Following microwave (head-focused) irradiation, perchloric acid extracts of rat brain regions (cortex, amygdala, and hippocampus) were analyzed for citrulline (determinant of NO) and high-energy phosphates (HEP) and their metabolites using high-performance liquid chromatograph (HPLC). KA-induced seizures produced a maximum increase in NO (3- to 6-fold) and a decrease in HEP (ATP 45-51% and phosphocreatine 45-58%) 2 h after KA injection in brain regions tested. 7-NI (50 mg/kg, i.p.) when given alone, reduced citrulline/NO levels (10 24%), while repeat administration of 7-NI (60 min apart) reduced NO levels by 32 49%. Neither application of 7-NI produced changes in HEP levels or toxicity. Pretreatment with 7-NI 30 min before KA injection, delayed the onset of seizures by 15-20 min, and significantly prevented an increase in NO and a decrease in HEP. Repeat administration of 7-NI, i.e. 30 min before and 30 min after KA injection, further increased protection by the delayed onset of seizures, attenuating the increase in NO and the decrease in HEP. Neurotoxicity of seizures involves activation of nNOS and of energy consumption in affected neurons. This increased energy consumption, coupled with decreased energy production caused by NO-induced mitochondrial dysfunction, may be a contributing factor to neuronal injury in KA toxicity. PMID- 12885441 TI - Sources of oxygen radicals in brain in acute ammonia intoxication in vivo. AB - The effects of acute ammonia intoxication on reactive oxygen species production by different sources in rat brain were studied. Ammonia intoxication in vivo leads to reduced activity of superoxide dismutase (SOD), catalase and glutathione peroxidase in brain nonsynaptic mitochondria and increased formation of O(2)(-) by submitochondrial particles. It also results in increased xanthine oxidase (XO) activity and decreased xanthine dehydrogenase (XDH)/XO activity ratio indicating conversion of XDH to XO and also increases monoamine oxidase A (MAO-A) activity but not of MAO-B. Blocking NMDA receptors with MK-801 prevents ammonia-induced oxidative stress, XDH to XO conversion and MAO-A activation. Ammonia intoxication did not lead to H(2)O(2) formation by mitochondria, in spite of increased O(2)(-) generation. The main source of H(2)O(2) in the mitochondrial matrix was Mn-SOD. Ammonia intoxication in vivo leads to increased superoxide and decreased hydrogen peroxide in nonsynaptic brain mitochondria. Increased superoxide is due to increased formation by the respiratory chain and by xanthine and aldehyde oxidases and decreased elimination by antioxidant enzymes. The reduced formation of hydrogen peroxide is due to the reduced activity of Mn-SOD. Prevention of ammonia-induced production of reactive oxygen species by MK-801 supports the idea that it is mediated by activation of NMDA receptors. PMID- 12885442 TI - Differential modulation of feline defensive rage behavior in the medial hypothalamus by 5-HT1A and 5-HT2 receptors. AB - Previous studies have established that the expression of defensive rage behavior in the cat is mediated over reciprocal pathways that link the medial hypothalamus and the dorsolateral quadrant of the midbrain periaqueductal gray matter (PAG). The present study was designed to determine the roles played by 5-HT(1A) and 5 HT(2C) receptors in the medial hypothalamus on the expression of defensive rage behavior elicited from electrical stimulation of the PAG. Monopolar stimulating electrodes were placed in the midbrain PAG from which defensive rage behavior could be elicited by electrical stimulation. During the course of this study, defensive rage was determined by measuring the latency of the "hissing" component of this behavior. Cannula-electrodes were implanted into sites within the medial hypothalamus from which defensive rage behavior could also be elicited by electrical stimulation in order that serotonergic compounds could be microinjected into behaviorally identifiable regions of the hypothalamus at a later time. Microinjections of the 5-HT(1A) receptor agonist 8-OHDPAT (0.1, 1.0 and 3.0 nmol) into the medial hypothalamus suppressed PAG-elicited hissing in a dose-dependent manner. Administration of the 5-HT(1A) antagonist p-MPPI (3.0 nmol) blocked the suppressive effects of 8-OHDPAT upon hissing. The suppressive effects of 8-OHDPAT were specific to defensive rage behavior because this drug (3 nmol) facilitated quiet biting attack. Microinjections of the 5-HT(2C) receptor agonist (+/-)-DOI hydrochloride into the medial hypothalamus (0.5, 1.0, and 3.0 nmol) facilitated the occurrence of PAG-elicited hissing in a dose-dependent manner. In turn, these facilitating effects were blocked by pretreatment with the selective 5-HT(2) antagonist, LY-53,857, which was microinjected into the same medial hypothalamic site. The findings of this study provide evidence that activation of 5-HT(1A) and 5-HT(2) receptors within the medial hypothalamus exert differential modulatory effects upon defensive rage behavior elicited from the midbrain PAG of the cat. PMID- 12885443 TI - Choroidal bodies: a new structure in the fourth ventricular choroid plexus of the rat and mouse. AB - Histologic study of the caudal end of the fourth ventricular choroid plexus of the rat and mouse revealed 1-4 small, discrete collections of cells that differed from the surrounding choroidal epithelial cells in appearance. They did not occur in other parts of the choroid plexuses. These choroidal bodies were not affected by a chemical toxin that caused hydropic degeneration of all the epithelial cells in the choroid plexuses. The function, if any, of the choroidal bodies is unknown. They were present in all rats that were studied by serial sections. PMID- 12885444 TI - Firing properties of frog tectal neurons in vitro. AB - Whole-cell recordings from frog tectal slices revealed different types of neuronal firing patterns in response to prolonged current injection. The patterns included regular spiking without adaptation, accelerating firing, adapting spiking, repetitive bursting and phasic response with only one spike. The observed firing patterns are similar to those found in the mammalian superior colliculus. The frog tectum could be a useful preparation in elucidating the relationship between neuronal function and membrane properties. PMID- 12885446 TI - Radiotherapy of small intracranial tumours with different advanced techniques using photon and proton beams: a treatment planning study. AB - BACKGROUND AND PURPOSE: The potential benefits and limitations of five different radiation techniques, 3D conformal radiotherapy (3DCRT), stereotactic arc therapy (SRS/T), intensity modulated radiotherapy with photons (IMRT), and radiotherapy with protons (spot scanning (SSp) or passive scattering (PSp)), have been assessed using comparative treatment planning methods in a cohort of patients presenting with 'benign' brain tumours. MATERIAL AND METHODS: Plans for five acoustic neurinomas, five meningiomas, and two pituitary adenomas were computed for all modalities using computed tomography (CT) scans to delineate planning target volume and organs at risk (OARs) and to predict dose distributions. Dose volume histograms were used for physical and simple biological evaluation. RESULTS: Proton techniques were shown to be superior to all photon approaches for the irradiation of small brain lesions in terms of target dose uniformity and conformity and in terms of sparing OARs. No major differences were observed between the results of the photon techniques, which were generally good for target coverage. Minimum target doses ranged from 81% with SRS/T to 93% with IMRT. The volume receiving more than 95% of the dose ranged from 95% (SRS/T) to 99% (PSp). No clear patterns of coverage dependence upon target shape were observed. Maximum brain stem irradiation ranged from 60% with IMRT to 26% with protons and the conformity index from 4.4 with IMRT to 2.5 with protons. Considering the rather long life expectancy of the patients suffering from meningiomas, neurinomas, and pituitary adenomas, the most important aspect to be considered, other than target coverage, is toxicity and in the long term, the possibility of secondary tumour induction. Considering these aspects, proton irradiation should be the irradiation technique of choice, when available. If not, IMRT, or even 3DCRT, techniques can provide an acceptable compromise, even without recurring to unconventional treatments like SRS/T, which require complex installations and high machine occupancy. PMID- 12885447 TI - Radiosurgery for brain metastasis: impact of CTV on local control. AB - PURPOSE: The purpose of the present analysis was to assess whether adding a 1 mm margin to the gross tumour volume (GTV) improves the control rate of brain metastasis treated with radiosurgery (RS). PATIENTS AND METHODS: All the patients had one or two brain metastases, 30 mm or less in diameter, and only one isocentre was used for RS. There were 23 females and 38 males. The median age was 54 years (34-76). The median Karnofsky performance status was 80 (60-100). At the time of RS, 23 patients had no evidence of extracranial disease and 38 had a progressive systemic disease. Thirty-eight patients were treated up-front with only RS. Twenty-three patients were treated for relapse or progression more than 2 months after whole brain radiotherapy. From January 1994 to July 1995, clinical target volume (CTV) was equal to GTV without any margin (33 metastases). From August 1995 to August 2000, CTV was defined as GTV plus a 1 mm margin (45 metastases). A dose of 20Gy was prescribed to the isocentre and 14Gy at the margin of CTV. RESULTS: The median follow-up was 10.5 months (1-45). The mean minimum dose delivered to GTV was 14.6Gy in the group without a margin and 16.8Gy in the group with a 1 mm margin (P<0.0001). The response of 11 metastases was never assessed because patients died before the first follow-up. Ten metastases recurred, eight in the group treated without a margin and two in the group treated with a 1 mm margin (P=0.01). Two-year local control rates were 50.7+/ 12.7% and 89.7+/-7.4% (P=0.008), respectively. Univariate analysis showed that the treatment group (P=0.008) and the tumour volume (P=0.009) were prognostic factors for local control. In multivariate analysis, only the treatment group with a 1 mm margin was an independent prognostic factor for local control (P=0.04, RR: 5.8, 95% CI [1.08-31.13]). There were no significant differences, either in overall survival rate or in early and late side effects, between the two groups. CONCLUSION: Adding a 1 mm margin to the GTV in patients treated with RS significantly improves the probability of metastasis control without increasing the side effects. PMID- 12885448 TI - A randomized trial comparing 35Gy in ten fractions with 60Gy in 30 fractions of cerebral irradiation for glioblastoma multiforme and older patients with anaplastic astrocytoma. AB - A randomized prospective clinical trial was conducted to compare conventional high dose radiotherapy with hypofractionated, short course radiotherapy in poor prognosis patients with high grade glioma. The primary endpoint was overall survival. PMID- 12885449 TI - Visualization of the corticospinal tract pathway using magnetic resonance axonography and magnetoencephalography for stereotactic irradiation planning of arteriovenous malformations. AB - Corticospinal tract (CST) information using anisotropic diffusion-weighted imaging and magnetoencephalography were integrated into radiosurgical planning for two patients with deeply seated arteriovenous malformation. The volume of CST receiving >10 Gy, >15 Gy, and maximum dose of CST could be reduced when plans were created with the aid of CST information compared with plans without the information. The results indicate that the use of CST information might reduce the risk of post-radiosurgical motor disturbance resulting from radiation necrosis. PMID- 12885450 TI - Factors determining acute normal tissue reactions during postoperative radiotherapy in endometrial cancer: analysis of 317 consecutive cases. AB - BACKGROUND AND PURPOSE: Acute radiotherapy reactions are commonly underestimated and under-reported in the literature. Our aim was to evaluate the incidence and risk factors for acute reactions during postoperative radiotherapy in endometrial cancer patients. MATERIAL AND METHODS: Performed was detailed retrospective analysis of 317 endometrial cancer patients given postoperative radiotherapy. Two hundred forty seven patients (78%) received both intracavitary (BRT) and external beam irradiation (EBRT), 49 patients (15%) received only BRT and 21 patients (7%) - only EBRT. BRT included radium (Ra) or cesium (Cs). The mean total dose at 0.5 cm for Ra and Cs was 50.5+/-10.3 Gy and 48.4+/-15.0 Gy, respectively, and the mean dose rate - 0.47+/-0.06 Gy/h and 1.42+/-0.41 Gy/h, respectively. Mean EBRT dose in the ICRU reference point was 49.0+/-3.7 Gy given in fractions of 1.54 2.49 Gy (mean 2.0+/-0.17 Gy). Radiotherapy and Oncology Group classification system was employed to score acute reactions. The impact of patient- and treatment-related factors on the risk of acute bowel and urinary bladder reactions was assessed with uni- and multivariate tests. RESULTS: Acute radiotherapy reactions of any grade occurred in 265 patients (84%) including bowel complications in 66% and urinary bladder complications in 36%. There were 21 severe (grade 3 or 4) reactions, all but one seen in the patients treated with combined EBRT and BRT. Higher total dose (P=0.024), higher EBRT dose (P=0.022) and higher age (P=0.026) were correlated with increased acute bowel toxicity in univariate analysis. Multivariate analysis showed that higher EBRT dose (P=0.015) and older age (P=0.016) were independently correlated with the risk of acute bowel events. Higher total dose (P=0.009), BRT dose (P=0.029), BRT dose rate (P=0.004), EBRT fraction size (P=0.007), the use of Cs BRT (P=0.001) and lower parity (P=0.041) were correlated with increased risk of acute bladder toxicity in univariate test. Multivariate analysis demonstrated that the independent risk factors for acute bladder events were BRT dose rate (P=0.002) and low parity (P=0.042) and there was a trend for EBRT dose (P=0.076). In multivariate analysis there was no impact of other clinical factors (FIGO stage, diabetes mellitus, hypertension, prior abdominal surgery) on the risk of acute bowel and/or bladder reactions nor was the impact of surgery-to-radiotherapy interval, overall radiotherapy time and overall treatment time. CONCLUSIONS: The risk of acute reactions depends both on treatment-related (BRT dose rate, EBRT dose) and patient-related factors (age, parity). Precise treatment prescription, planning and verification are of paramount concern. Further studies are warranted to evaluate the impact of extrinsic and intrinsic factors associated with acute normal tissue injury. PMID- 12885451 TI - Radiation therapy of the pelvic and paraaortic lymph nodes in cervical carcinoma: a prospective three-dimensional analysis of patient positioning and treatment technique. AB - PURPOSE: Pelvic box fields in prone position are the standard treatment for patients with cervical carcinomas. The issue investigated in this report is whether this technique should also be used when extending the planning target volume to the paraaortic region. MATERIALS AND METHODS: In a prospective study of eight consecutive patients with cervical carcinomas, two patient positions (prone and supine) and three radiation techniques (A, anteroposterior/posteroanterior opposed fields; B, four-field box; and C, three-field technique) were examined concerning the dose to critical organs. The analysis was based on three dimensional planning, dose-volume histograms and normal tissue complication probabilities (NTCP). RESULTS: Compared to the prone position, the supine position led to improved organ sparing in four of seven organs (liver, both kidneys, spinal canal). In two of seven organs (rectum and bladder) no difference between prone and supine position was observed. The best sparing of small bowel was achieved in prone position. Technique B followed by technique C in the supine position resulted in the best overall sparing of critical organs concerning the volumes receiving the respective TD(5/5) doses or more. Mean NTCP values for liver, rectum and bladder were below 1.0%. The highest values of up to 12% were found for both kidneys in prone position with C and for the spinal canal with A in the prone and supine position. CONCLUSION: According to this analysis, for the treatment of the pelvic and paraaortic lymph node regions together, supine position and technique B (alternatively C) should be preferred despite the advantages of prone position on belly boards for pelvic irradiation alone. PMID- 12885452 TI - The impact of sectional imaging on dose escalation in endocavitary HDR brachytherapy of cervical cancer: results of a prospective comparative trial. AB - PURPOSE: The purpose of this comparative prospective study was to assess the effect of CT and MR based individualisation and adaptation on the dose distribution in the target volume and organs at risk compared to a radiography based procedure. MATERIAL AND METHODS: In 15 patients MR scans, in 10 patients additional axial CT-scans with compatible tube-ring applicator in situ were performed and digitally transferred to the PLATO(R) planning system. Considering clinical examination and MR-scan before radiotherapy individual 3-D dose distribution was calculated and adapted based on (1) two orthogonal radiographs; (2) isodoses superimposed on the CT images; and (3) isodoses superimposed on the MR images. Adaptation was strictly limited by the dose level at 2 cm(3) bladder or rectum volume (D(2)) to allow comparison of CT and MR plans. All three individualised dose distributions were superimposed on the MR images and cumulative dose-volume histograms were calculated for comparison. RESULTS: 3-D individualisation based on sectional imaging enabled higher dose to the target volume (isodose enclosing 95% of the CTV=CTV(95)) compared to individualised treatment plans based on orthogonal radiographs by a mean factor of 1.2 (1-1.7). The dose to bladder and/or rectum wall was at the same time not increased beyond the prescribed tolerance level (71% of the prescribed target dose). In a subgroup of 10 patients MRI based treatment plans were superior to CT based treatment plans allowing for a higher dose (138% vs. 124%). CONCLUSION: Sectional imaging based treatment planning, in particular using MR, was superior to radiography allowing for a clinical meaningful dose escalation without increasing the dose to bladder and rectum beyond the tolerance level. PMID- 12885453 TI - Pelvic insufficiency fractures in postmenopausal woman with advanced cervical cancer treated by radiotherapy. AB - PURPOSE: To assess the predisposing factors and clinical characteristics of pelvic insufficiency fractures (PIF) in postmenopausal women with pelvic irradiation. MATERIAL AND METHODS: A total 335 postmenopausal patients with cervical cancer of the intact uterus treated with radiation therapy between 1983 and 1998 were reviewed. Total external dose was delivered between 45 and 50.4 Gy with parallel opposed anteroposterior portals. Total brachytherapy dose at point A was delivered between 10 and 36 Gy. PIF were diagnosed by bone scintigraphy and confirmed by computed tomography. The cumulative incidence of symptomatic PIF was estimated by actuarial methods. Potential risk factors (age, weight, type II diabetes, delivery, menopause, total external dose, total brachytherapy dose) were assessed. RESULTS: Fifty-seven (17.0%) of 335 patients were diagnosed as having PIF. Forty-seven patients were symptomatic and ten were asymptomatic. Parameters carrying a significant association with PIF were body weight 49 kg or below (P=0.044) in stepwise logistic regression analysis. The cumulative incidence of symptomatic PIF at 5 years was 17.9% calculated by the Kaplan-Meier method. A body weight of 49 kg or below and more than three deliveries were identified as having a significant effect on symptomatic PIF in univariate analysis (P=0.021, P=0.003, log-rank test) and Cox life table regression analysis (P=0.038, P=0.013). Five patients required narcotic agents and eight patients required hospital admission. CONCLUSIONS: We should consider reducing the dose contribution to the sacrum and sacroilac joints, without underdosing the tumor, especially in postmenopausal women with many deliveries or low body weight. PMID- 12885454 TI - Respiration-induced movement of the upper abdominal organs: a pitfall for the three-dimensional conformal radiation treatment of pancreatic cancer. AB - Respiration-induced movement of the upper abdominal organs (pancreas, liver and kidneys) was assessed in 12 subjects using dynamic magnetic resonance imaging. The movement of each organ in the cranio-caudal, the lateral and the anterior posterior direction was deduced from the movement of the center of gravity on two dimensional images. This center of gravity was computed from the volume delineated on sequential 8-mm slices of both sagittal and coronal dynamic series. The largest movements were noticed in the cranio-caudal direction for pancreas and liver (23.7+/-15.9 mm and 24.4+/-16.4 mm). The kidneys showed smaller movements in the cranio-caudal direction (left kidney 16.9+/-6.7 mm and right kidney 16.1+/-7.9 mm). The movements of the different organs in the anterior posterior and lateral directions were less pronounced. It is of the greatest importance to be aware of these movements in the planning of a conformal radiation treatment for pancreatic cancer. PMID- 12885455 TI - Concomitant boost radiotherapy for muscle invasive bladder cancer. AB - PURPOSE: To evaluate the feasibility and efficacy of a concomitant partial bladder boost schedule in radiotherapy for invasive bladder cancer, coupling a limited boost volume with shortening of the overall treatment time. METHODS AND MATERIALS: Between 1994 and 1999, 50 patients with a T2-T4 N0M0 transitional cell carcinoma of the bladder received radiotherapy delivered in a short overall treatment time with a concomitant boost technique. With this technique a dose of 40 Gy in 2-Gy fractions was administered to the small pelvis with a concomitant boost limited to the bladder tumor area plus margin of 15 Gy in fractions of 0.75 Gy. The total tumor dose was 55 Gy in 20 fractions in 4 weeks. Toxicity was scored according to EORTC/RTOG toxicity criteria. RESULTS: The feasibility of the treatment was good. Severe acute toxicity >/=G3 was observed in seven patients (14%). Severe late toxicity >/=G3 was observed in six patients (13%). Thirty seven patients (74%) showed a complete and five (10 %) a partial remission after treatment. The actuarial 3-year freedom of local progression was 55%. CONCLUSION: In external radiotherapy for muscle invasive bladder cancer a concomitant boost technique coupling a partial bladder boost with shortening of the overall treatment time provides a high probability of local control with acceptable toxicity. PMID- 12885456 TI - Comparison of megavoltage position verification for prostate irradiation based on bony anatomy and implanted fiducials. AB - PURPOSE: The patient position during radiotherapy treatment of prostate cancer can be verified with the help of portal images acquired during treatment. In this study we quantify the clinical consequences of the use of image-based verification based on the bony anatomy and the prostate target itself. PATIENTS AND METHODS: We analysed 2025 portal images and 23 computed tomography (CT) scans from 23 patients with prostate cancer. In all patients gold markers were implanted prior to CT scanning. Statistical data for both random and systematic errors were calculated for displacements of bones and markers and we investigated the effectiveness of an off-line correction protocol. RESULTS: Standard deviations for systematic marker displacement are 2.4 mm in the lateral (LR) direction, 4.4 mm in the anterior-posterior (AP) direction and 3.7 mm in the caudal-cranial direction (CC). Application of off-line position verification based on the marker positions results in a shrinkage of the systematic error to well below 1 mm. Position verification based on the bony anatomy reduces the systematic target uncertainty to 50% in the AP direction and in the LR direction. No reduction was observed in the CC direction. For six out of 23 patients we found an increase of the systematic error after application of bony anatomy-based position verification. CONCLUSIONS: We show that even if correction based on the bony anatomy is applied, considerable margins have to be set to account for organ motion. Our study highlights that for individual patients the systematic error can increase after application of bony anatomy-based position verification, whereas the population standard deviation will decrease. Off-line target-based position verification effectively reduces the systematic error to well below 1 mm, thus enabling significant margin reduction. PMID- 12885457 TI - Osteosarcomas occurring as second malignant neoplasms. PMID- 12885458 TI - Early radiotherapy related morbidity. PMID- 12885459 TI - Updates on the cytogenetics and molecular genetics of bone and soft tissue tumors: osteosarcoma and related tumors. PMID- 12885460 TI - Cytogenetic and molecular analysis of an unusual case of acute promyelocytic leukemia with a t(15;17;17)(q22;q23;q21). AB - We present a 52-year-old female with a clinical history of acute myelocytic leukemia, probable acute promyelocytic leukemia (APL). Flow cytometry results were somewhat unusual. Specifically, the promyelocytic population showed partial positivity for antigens not usually expressed in APL (HLA-DR and CD117). The interpretation of these results was that the abnormal population contained a proportion of very early promyeolocytes that had not completely lost all their "precursor" antigens. Cytogenetic analysis of a bone marrow aspirate showed a t(15:17;17)(q22;q23;q21) in all cells analyzed. Fluorescence in situ hybridization (FISH) analysis using the PML-RARA DNA probe showed a positive signal pattern (fusion) in 100% of 200 total interphase and metaphase cells examined, confirming the presence of the PML-RARA rearrangement. Multicolor FISH, which produces 24 colors to differentiate all chromosomes in a single hybridization, was applied. This study confirmed the cytogenetic interpretation of the rearrangement. No material from any other chromosome was detected on the second smaller derivative chromosome 17. Additional studies using the RARA(17q21) break-apart DNA FISH probe showed that 17q21 (RARA) was not rearranged on the derivative chromosome 17 that received the q22-->qter segment from chromosome 15. The RARA locus on the smaller derivative 17 was the allele involved in the fusion in this three-way rearrangement. The signal pattern was consistent in 100% of interphase and metaphase cells scored. This unusual t(15;17;17) prompted us to investigate further using reverse-transcription polymerase chain reaction with primers from the 3' and 5' regions of both the RARA and PML loci. These studies showed that the PML-RARA fusion was present, but the complementary fusion RARA PML, which is usually detectable, was absent. The patient is responding well to standard treatment protocols. PMID- 12885461 TI - Telomeric fusion as a mechanism for the loss of 1p in meningioma. AB - Characteristic cytogenetic aberrations are found in the various histopathological designations of meningioma. These aberrations range from the loss of 22q in histologically benign tumors to complex hypodiploid karyotypes in atypical and malignant tumors. This progression is characterized by increasing chromosome loss and instability, with a critical step being the loss of 1p. We report a detailed cytogenetic investigation of chromosome aberrations in a series of 88 meningiomas using Giemsa banding and multicolor spectral karyotyping (SKY). Clonal chromosome aberrations were identified in 46 (52%) tumors by G banding. Thirty-five tumors showing complex chromosome aberrations not fully characterized by G banding were subsequently reanalyzed by SKY. The SKY technique refined the G-band findings in 18 (51%) of the tumors on which it was applied. The most common features of cytogenetic progression in the complex karyotypes were chromosome arm-specific losses relating to the formation of deletions and dicentric chromosomes involving 1p. Part or all of 1p was lost in 19 tumors. Five tumors showed evidence for the loss of 1p in a progressive step-wise series of telomeric fusions involving the formation of unstable intermediates. Five recurring dicentric chromosomes were identified, including dic (1;11)(p11;p11), dic(1;12)(p12 approximately p13;p11), dic(1;22)(p11;q12 approximately q13), dic(7;19)(p11;p11), and dic(19;22)(p11 approximately p13;q11 approximately q13). These findings provide evidence that telomeric fusions play a role in the formation of clonal deletions, dicentrics, and unbalanced translocations of 1p. The loss of 1p has possible diagnostic and prognostic implications in the management of meningioma. PMID- 12885462 TI - Tetraploid acute promyelocytic leukemia with double t(15;17) and PML/RARA rearrangements detected by fluorescence in situ hybridization analysis. AB - Acute promyelocytic leukemia (APL) is characterized by a serious hemorrhagic syndrome, unique morphologic findings, and its response to retinoids. Tetraploidy is a very rare chromosomal abnormality in acute myelocytic leukemia. This report presents a unique case of APL with a tetraploid clone characterized by two t(15;17) without other chromosomal changes, as well as PML/RARA rearrangements confirmed fluorescence in situ hybridization. The morphology of the blast cells was that of the classic M3 subtype, but the mean blast size exceeded that of control APL cases with diploidy. A chromosomal study revealed a 92,XXXX,t(15;17)(q22;q21)x2 karyotype in all 20 metaphase spreads. Despite all trans-retinoic acid (ATRA) treatment and chemotherapy, leukemic cells persisted in the blood, and the patient died of an intracranial hemorrhage on the 16th day after admission. PMID- 12885463 TI - Rearrangement of the MLL gene and a region proximal to the RARalpha gene in a case of acute myelocytic leukemia M5 with a t(11;17)(q23;q21). AB - A case of acute myelocytic leukemia (AML) M5 subtype (French-American-British classification), in a 13-year-old girl showed the abnormal karyotype 46,XX,t(11;17)(q23;q21) in all bone marrow cells analyzed. Rearrangements involving 11q23 are frequent in cases of AML M5 and often involve the MLL gene. Nevertheless, t(11;17)(q23;q21) is very rare in this type of leukemia. In acute promyelocytic leukemia, the RARalpha gene, located at 17q21, is involved in almost all cases. Fluorescence in situ hybridization studies revealed a deletion of the C-terminal part of the MLL gene and a translocation of the RARalpha gene on the derivative chromosome 11, proximal to the remaining part of the MLL gene. However, hybridization with the LSI RARA dual color break-apart rearrangement probe showed that the RARalpha gene was not rearranged in this translocation. This is the first study reporting a t(11;17)(q23;q21) with a deletion distal to MLL gene exon 6 in a case of AML M5. Furthermore, this is the second study that strongly suggests the implication of a gene proximal and close to the RARalpha locus in a case of AML M5. According to these results, the discovery of new fusion partner genes of MLL and the precise characterization of t(11;17) will be important for the understanding of neoplastic cell differentiation in AML M5. PMID- 12885464 TI - A Li-Fraumeni syndrome family with retained heterozygosity for a germline TP53 mutation in two tumors. AB - We identified a missense germline mutation (Gly245Ser) in one of the mutation hot spots of the TP53 gene in two affected members of a Li-Fraumeni syndrome family. We also analyzed their tumors, a liposarcoma and a colorectal carcinoma. Both tumors exhibited p53 protein accumulation but none of them showed loss of the wild-type allele of the TP53 gene. We reviewed all published cases of tumors in germline TP53 mutation carriers where loss of heterozygosity data were available and identified 84 tumors with loss of the wild-type allele, 57 tumors with retention of heterozygosity, and 9 tumors with loss of the allele harboring the germline mutation. Among the tumors showing p53 accumulation, we observed a significant difference in the fraction of tumors showing p53 protein accumulation between the tumors with loss of the wild-type allele and those with retention of TP53 heterozygosity. This supports the idea that the pathogenesis of tumors in germline TP53 mutation carriers does not have to be associated with loss of the wild-type TP53 allele. The product of the normal allele can potentially be inactivated by a variety of other mechanisms or, as suggested by the analysis, many of these tumors may even preserve the activity of the wild-type p53 protein. PMID- 12885465 TI - Three new cases of non-Hodgkin lymphoma with t(9;14)(p13;q32). AB - The majority of non-Hodgkin lymphomas of B-cell type (B-NHL) exhibit chromosomal abnormalities including many types of reciprocal translocations closely related to specific histopathologic entities. The t(9;14)(p13;q32) has been recognized as a primary genetic event directly involved in the development of lymphoplasmacytic lymphoma. In the 14 published cases, the t(9;14)(p13;32) seems to delineate a variety of low-grade B-cell disorders characterized by a common clinical history and immunopathologic similarities. We report here three new cases presenting a t(9;14)(p13;q32) with other chromosomal abnormalities which have been referred to as B-cell low-grade or high-grade malignant lymphoproliferative disorders. Two of these cases showed diffuse large B cell lymphoma morphology and two patients had a favorable clinical outcome. These data suggest that t(9;14)(p13;q32) is not restricted to low-grade lymphoma. PMID- 12885466 TI - Allelic imbalance in colorectal cancer at the CRAC1 locus in early-onset colorectal cancer. AB - A susceptibility gene to colorectal adenomas and carcinoma (CRAC1) on chromosome region 15q14 approximately q22 has been proposed on the basis of linkage in a single family. Allele-specific loss of heterozygosity (LOH) in tumors of affected family members suggests that the causative gene functions as a tumor-suppressor gene. The genes that are mutated in inherited cancer syndromes are often involved in the pathogenesis of sporadic cancer. To determine whether CRAC1 plays a role in colorectal carcinogenesis in general, we have studied 277 cases of early-onset colorectal cancers for allele loss at 15q14 approximately q22 using four microsatellite markers (D15S970, D15S117, D15S971, and D15S1028) that define the region of maximal linkage. The frequency of LOH detected was between 14% and 22%, but there was no significant association between LOH at each adjacent marker. Most cancers caused by loss of expression of a tumor suppressor involve large scale deletion of one allele. On this basis, our findings suggest that CRAC1 is unlikely to be implicated in the development of colorectal cancer in general or, if involved, it is through small somatic mutations or other loss of function mechanisms rather than allele loss. PMID- 12885467 TI - Insertion of chromosome 11 in chromosome 4 resulting in a 5'MLL-3'AF4 fusion gene in a case of adult acute lymphoblastic leukemia. AB - We report on a 69-year-old woman with B-lineage acute lymphoblastic leukemia. Cytogenetic studies at diagnosis with R banding showed an insertion, ins(4;11)(q21;q13q23). Fluorescence in situ hybridization (FISH) with whole chromosome painting probes confirmed the insertion of chromosome 11 material into chromosome 4. FISH using the MLL probe showed the translocation of the 5' end of MLL into chromosome 4. Since the 5'MLL-3'AF4 fusion transcript was detected by a reverse transcriptase polymerase chain reaction, we concluded that the insertion of part of chromosome 11 split the AF4 gene in two, resulting in the presence of the 5'MLL-3'AF4 fusion gene on the der(4) instead of the der(11), as commonly observed. Our findings stress the value of combining banding cytogenetics with FISH and molecular techniques to better assess rearrangements in leukemia. PMID- 12885468 TI - Mosaic trisomy 2 in myelodysplastic syndromes and acute myeloblastic leukemias. AB - We present a short report here of two more patients with trisomy 2 as the sole chromosomal abnormality in a hematologic malignancy. Although trisomy 2 is a recognized abnormality in neoplasms, particularly hepatoblastomas, to the best of our knowledge only three other cases have been reported with trisomy 2, in patients with a hematologic malignancy. The two cases presented here of myelodysplastic syndrome transforming to acute myeloblastic leukemia and chronic myelomonocytic leukemia showed trisomy 2 as the sole abnormality. PMID- 12885469 TI - Constitutional pericentric inversion of chromosome 9 and acute leukemia. AB - In view of the recent reports demonstrating delayed engraftment after autologous and allogeneic transplantation from donors with constitutional pericentric inversion of chromosome 9, [inv(9)], we conducted a retrospective study on six patients with acute leukemia and inv(9) to investigate if there is an impaired engraftment potential of the inv(9) hematopoietic stem cells. All but one of our patients had poor outcome. The hematopoietic recovery after induction chemotherapy was not prolonged. It is possible that the hematopoietic defects of inv(9) become more apparent after repeated courses of chemotherapy. Alternatively, the number of patients in our series may have been too small to detect a partial hematopoietic defect in patients with constitutional inv(9). Larger studies are required to confirm our findings. PMID- 12885470 TI - Trisomy 6 acquired in lymphoid blast transformation of chronic myelocytic leukemia with t(9;22). PMID- 12885471 TI - Translocation (8;20), a variant of t(8;21), in T-acute lymphoblastic leukemia: a first report. PMID- 12885472 TI - WHO 2003-08: a programme of quiet thunder takes shape. PMID- 12885473 TI - Mathematical morbidity in paediatric intensive care. PMID- 12885474 TI - New precision in measuring trachoma infection. PMID- 12885475 TI - Dietary assessment and the reliability of nutritional epidemiology reports. PMID- 12885476 TI - Breast-cancer prevention: is the risk-benefit ratio in favour of tamoxifen? PMID- 12885477 TI - Dam medicine. PMID- 12885478 TI - Mortality in relation to oral contraceptive use and cigarette smoking. AB - BACKGROUND: As part of the Oxford Family Planning Association study, we compared mortality in relation to oral contraceptive use and smoking to highlight the differences between them from the perspective of public health. METHODS: The study consisted of 17032 women, aged 25-39 years at entry, recruited between May 1, 1968, and July 31, 1974, who had used oral contraceptives, a diaphragm, or an intrauterine device. We assessed mortality from follow-up data recorded until Dec 31, 2000. The analysis is based on woman-years of observation. FINDINGS: We analysed 889 deaths. Women who had ever used oral contraceptives had increased mortality from cervical cancer (rate ratio 7.2, 95% CI 1.1-303), and decreased mortality from other uterine (0.2, 0.0-0.8) and ovarian cancers (0.4, 0.2-0.7). Oral contraceptives had some adverse effect on deaths from ischaemic heart disease in women who smoked 15 or more cigarettes per day. For all causes of mortality, the rate ratio for death in women who ever used oral contraceptives was 0.89 (95% CI 0.77-1.02). By contrast, this rate ratio was 1.24 (1.03-1.49) in those who smoked one to 14 cigarettes per day, and 2.14 (1.81-2.53) in those who smoked 15 or more cigarettes per day. INTERPRETATION: There was no harmful effect of oral contraceptive use on overall mortality. By contrast, death from all causes was more than twice as high in smokers of 15 or more cigarettes a day as in non-smokers. The harmful effect was already apparent in women aged 35-44 years. PMID- 12885479 TI - Validation of the paediatric logistic organ dysfunction (PELOD) score: prospective, observational, multicentre study. AB - BACKGROUND: Multiple organ dysfunction syndrome is more frequent than death in paediatric intensive care units. Estimation of the severity of this syndrome could be a useful additional outcome measure in clinical trials in such units. We aimed to validate the paediatric logistic organ dysfunction (PELOD) score and estimate its validity when recorded daily (dPELOD). METHODS: We did a prospective, observational, multicentre cohort study in seven multidisciplinary, tertiary-care paediatric intensive care units of university-affiliated hospitals (two French, three Canadian, and two Swiss). We included 1806 consecutive patients (median age 24 months; IQR 5-90). PELOD score includes six organ dysfunctions and 12 variables and was recorded daily. For each variable, the most abnormal value each day and during the whole stay were used in calculating the dPELOD and PELOD scores, respectively. Outcome was vital status at discharge. We used Hosmer-Lemeshow goodness-of-fit tests to evaluate calibration and areas under receiver operating characteristic curve (AUC) to estimate discrimination. FINDINGS: 370 (21%) patients had no organ dysfunction, 471 (26%) had one, 457 (25%) had two, and 508 (28%) had three or more. Case fatality rate was 6.4% (115 deaths). PELOD score was significantly higher in non-survivors (mean 31.0 [SE 1.2]) than survivors (9.4 [0.2]; p<0.0001). Calibration (p=0.54) and discrimination (AUC=0.91, SE=0.01) of PELOD and dPELOD (p> or =0.39; AUC> or =0.79) scores were good. INTERPRETATION: PELOD and dPELOD scores are valid outcome measures of the severity of multiple organ dysfunction syndrome in paediatric intensive care units; their use should significantly reduce the sample size required to complete clinical trials in critically ill children. PMID- 12885480 TI - Lead lines. PMID- 12885481 TI - Strategies for control of trachoma: observational study with quantitative PCR. AB - BACKGROUND: Antibiotics are an important part of WHO's strategy to eliminate trachoma as a blinding disease by 2020. At present, who needs to be treated is unclear. We aimed to establish the burden of ocular Chlamydia trachomatis in three trachoma-endemic communities in Tanzania and The Gambia with real-time quantitative PCR. METHODS: Conjunctival swabs were obtained at examination from 3146 individuals. Swabs were first tested by the qualitative Amplicor PCR, which is known to be highly sensitive. In positive samples, the number of copies of omp1 (a single-copy C trachomatis gene) was measured by quantitative PCR. FINDINGS: Children had the highest ocular loads of C trachomatis, although the amount of pooling in young age groups was less striking at the site with the lowest trachoma frequency. Individuals with intense inflammatory trachoma had higher loads than did those with other conjunctival signs. At the site with the highest prevalence of trachoma, 48 of 93 (52%) individuals with conjunctival scarring but no sign of active disease were positive for ocular chlamydiae. INTERPRETATION: Children younger than 10 years old, and those with intense inflammatory trachoma, probably represent the major source of ocular C trachomatis infection in endemic communities. Success of antibiotic distribution programmes could depend on these groups receiving effective treatment. PMID- 12885482 TI - Survivin and molecular pathogenesis of colorectal cancer. AB - BACKGROUND: Colorectal cancer is thought to originate in the expansion of colonic crypt cells as a result of aberrant gene expression caused by transcription factors of the T-cell factor (TCF)/beta-catenin family. Survivin is a bifunctional regulator of cell death and cell proliferation expressed during embryonic development but undetectable in healthy adult tissues and re-expressed in many cancers, including colorectal cancer. METHODS: We investigated gene expression by promoter analysis, mutagenesis, and electrophoretic mobility shift assay in colorectal cancer cells. Survivin expression in human and mouse embryonic intestine was determined by in-situ hybridisation and immunohistochemistry. Changes in apoptosis were monitored in cell lines engineered to express stabilising mutations in beta catenin. FINDINGS: TCF/beta catenin stimulated a six-fold to 12-fold increased expression of the survivin gene in colorectal cancer cells. Three TCF-binding elements (TBE) in the survivin promoter were occupied by nuclear factors in colorectal cancer cells, and mutagenesis of the two proximal TBE sites abolished survivin gene expression by 75-79%. Strongly expressed at the bottom of human and mouse embryonic intestinal crypts, expression of survivin was lost in TCF-4 knockout animals, and a TCF-4 dominant negative mutant blocked survivin gene transcription in colorectal cancer cells. Expression of non-destructible beta catenin mutants increased survivin expression and protected against ultraviolet-B-induced apoptosis. INTERPRETATION: Stimulation of survivin expression by TCF/beta catenin might impose a stem cell like phenotype to colonic crypt epithelium coupling enhanced cell proliferation with resistance to apoptosis, and contribute to the molecular pathogenesis of colorectal cancer. PMID- 12885483 TI - HIV-1 transmission during scintigraphy. PMID- 12885484 TI - Haemoglobin concentration and prognosis in new cases of heart failure. AB - Anaemia is common in severe chronic heart failure and is reported to be a predictor of death. We investigated 552 patients (median age 76 years, range 29 95; 54% men [n=296]), in whom the duration of heart failure was sufficiently short that it would be unlikely to affect haemoglobin concentrations. By contrast with studies in established chronic heart failure, haemoglobin was not independently associated with prognosis when age and serum creatinine concentration were included in the analysis. The adverse effects of anaemia on survival might be a consequence of chronic heart failure rather than a separate process causing disease progression. PMID- 12885485 TI - Are imprecise methods obscuring a relation between fat and breast cancer? AB - Pooled analyses of cohort studies show no relation between fat intake and breast cancer risk. However, food-frequency questionnaire (FFQ) methods used in these studies are prone to measurement error. We assessed diet with an FFQ and a detailed 7-day food diary in 13070 women between 1993 and 1997. We compared 168 breast-cancer cases incident by 2000 with four matched controls. Risk of breast cancer was associated with saturated-fat intake measured with the food diary (hazard ratio 1.22 [95% CI 1.06-1.40], p=0.005, per quintile increase in energy adjusted fat intake), but not with saturated fat measured with the FFQ (1.10 [0.94-1.29], p=0.23). Dietary measurement error might explain the absence of a significant association between dietary fat and breast-cancer risk in cohort studies. PMID- 12885486 TI - Trachoma. AB - Trachoma is the most common infectious cause of blindness. It is caused by ocular serovars of Chlamydia trachomatis. Transmission is favoured in poor communities, where crowding is common and access to water and sanitation inadequate. Repeated reinfection over many years causes dense scarring of the upper eyelid. The resultant inversion of the lashes abrades the eyeball, and the abrasion leads to corneal opacification and visual impairment. The host immune response is probably at least partly the cause of this process. The "SAFE" strategy is used for the control of trachoma: surgery for in-turned lashes, antibiotics for active disease, facial cleanliness, and environmental improvement. The demonstration that a single oral dose of the antibiotic azithromycin is as effective as 6 weeks of topical tetracycline was an important advance in trachoma control. By means of the SAFE strategy, WHO and its partners aim to eliminate trachoma as a public health problem by the year 2020. PMID- 12885487 TI - Non-steroidal anti-inflammatory drugs and molecular carcinogenesis of colorectal carcinomas. AB - CONTEXT: Colorectal cancer is the second most common cause of cancer-related mortality in the west. The high incidence and mortality make effective prevention an important public-health and economic issue. Non-steroidal anti-inflammatory drugs (NSAIDs) can inhibit colorectal carcinogenesis and are among the few agents known to be chemopreventive. Randomised trials have shown that sulindac and celecoxib suppress the development of adenomatous polyps and cause regression of existing polyps in patients with familial adenomatous polyposis (FAP), who have a high risk for developing colorectal cancer. The mechanisms by which NSAIDs inhibit neoplastic growth are not fully known. STARTING POINT: Two recently reported randomised placebo-controlled trials show a chemopreventive effect of aspirin in populations other than those with FAP (Robert Sandler and colleagues, N Engl J Med 2003; 348: 883-90; John Baron and colleagues, N Engl J Med 2003; 348: 891-99). In the Sandler study 635 patients with colorectal cancer were randomised to receive 325 mg aspirin or placebo daily. After a follow-up of around 31 months, the mean number of adenomas was lower in the aspirin group than in the placebo group, corresponding to a relative risk of any recurrent adenoma in the aspirin group of 0.65. In the Baron study 1121 patients with colorectal adenomas were assigned to receive 81 or 325 mg aspirin or placebo daily. Follow up colonoscopy, 32 months after the index endoscopy, showed an incidence of one or more adenomas of 38% in the 81 mg aspirin group, 45% in the 325 mg aspirin group, and 47% in the placebo group. Together, these studies indicate a moderate chemopreventive effect of aspirin in populations with an intermediate risk of developing colorectal cancer. WHERE NEXT? The anticancer properties of NSAIDs have been demonstrated in vitro and in animal studies, epidemiological reports, and intervention studies. Several mechanisms through which NSAIDs alter colonic carcinogenesis have been elucidated, including the induction of apoptosis in neoplastic cells, via mechanisms dependent and independent of cyclo-oxygenase. Some studies have suggested an important role for the cell-cycle regulating protein p21 in mediating the chemopreventive effect of sulindac. A decrease in p21 expression may be one of the main oncogenic events in the development of colorectal cancer. Thus p21 could be the molecular link in the chemopreventive effects of NSAIDs. PMID- 12885488 TI - Applying an equity lens to child health and mortality: more of the same is not enough. AB - Gaps in child mortality between rich and poor countries are unacceptably wide and in some areas are becoming wider, as are the gaps between wealthy and poor children within most countries. Poor children are more likely than their better off peers to be exposed to health risks, and they have less resistance to disease because of undernutrition and other hazards typical in poor communities. These inequities are compounded by reduced access to preventive and curative interventions. Even public subsidies for health frequently benefit rich people more than poor people. Experience and evidence about how to reach poor populations are growing, albeit largely through small-scale case studies. Successful approaches include those that improve geographic access to health interventions in poor communities, subsidized health care and health inputs, and social marketing. Targeting of health interventions to poor people and ensuring universal coverage are promising approaches for improvement of equity, but both have limitations that necessitate planning for child survival and effective delivery at national level and below. Regular monitoring of inequities and use of the resulting information for education, advocacy, and increased accountability among the general public and decision makers is urgently needed, but will not be sufficient. Equity must be a priority in the design of child survival interventions and delivery strategies, and mechanisms to ensure accountability at national and international levels must be developed. PMID- 12885489 TI - Precious adults: a lesson in grown-up congenital heart disease. PMID- 12885490 TI - Clinical trials, genetic add-ons, and the question of benefit-sharing. AB - Discussion of benefit-sharing has become common in the sphere of human genetic research. Roughly, this term means that individuals or organisations who could reap financial rewards from research into human genetics have some obligation to share the benefits of this research, perhaps with the people who made the research possible in the first place, or with humankind more broadly. This idea has met with some acceptance, finding its way into policy recommendations and statements of several prominent groups. However, the issue of benefit-sharing is generally raised in the context of large-scale population-based genetic studies. Other sources of human DNA are often ignored. In particular, little attention has been paid to the increasingly common practice of collecting genetic samples as add-ons to clinical drug trials. Generally such trials do not specify a use for these samples, which are collected for purposes of potential future research. We argue that if a case for benefit-sharing can be made for genetic studies in general, it can be made for add-on studies as well. We suggest some ways in which benefit-sharing might be implemented for genetic add-on studies. PMID- 12885491 TI - Mortality reduction by breast-cancer screening. PMID- 12885492 TI - Mortality reduction by breast-cancer screening. PMID- 12885493 TI - The UK breast-screening programme should start at age 47 years. PMID- 12885494 TI - Mortality reduction by breast-cancer screening. PMID- 12885495 TI - Should we encourage exclusive breastfeeding at all cost? PMID- 12885496 TI - Clitoral surgery and sexual outcome in intersex individuals. PMID- 12885497 TI - Economists tell scientists AIDS drug projects can be scaled up. Calls for more money dominate the agenda but economists say more can be done with current funds. PMID- 12885498 TI - The need to update the classification of acute malnutrition. PMID- 12885499 TI - Regulatory assessment of new drugs in developing countries. PMID- 12885500 TI - Preimplantation genetic diagnosis. PMID- 12885501 TI - The real cost of an affordable vaccine for meningococcus A. PMID- 12885502 TI - Brazil offers expertise and support to Africa. PMID- 12885503 TI - What is the WMA for? The case of the Israeli Medical Association. PMID- 12885504 TI - Bush's visit to Uganda raises hopes and sparks controversy. PMID- 12885505 TI - Cause of dosing errors. PMID- 12885506 TI - FDA says it will require trans-fat amounts on food labels. PMID- 12885507 TI - Placebos in clinical care: for whose pleasure? PMID- 12885508 TI - Improve tuberculosis services for HIV patients, says WHO. PMID- 12885509 TI - Codex adopts new standards on GM foods, irradiation, and animal feed. PMID- 12885510 TI - Bans and proscriptions. PMID- 12885511 TI - Invasion of the managers. PMID- 12885512 TI - Study of interaction of poly(1-vinyl-2-pyrrolidone) with a surface of highly dispersed amorphous silica. AB - Interaction of poly(1-vinyl-2-pyrrolidone) with the surface of highly dispersed silica and character of its thermal destruction were investigated by means of ad desorption measurement, vibration and electron spectroscopy, and thermal gravimetric analysis with programmed heating. The correlation between the character of polymer keeping on the surface of silicon dioxide and its thermal decomposition was established. PMID- 12885513 TI - A potentiometric study of the adsorption of a cationic surfactant onto laponite in water-methanol and water-dimethylsulfoxide mixtures. AB - We studied the adsorption of a cationic surfactant, dodecyltrimethylammonium bromide (DTABr), onto laponite in water-methanol and water-DMSO mixtures at 298 K. Adsorption isotherms were determined by potentiometric methods using DTA(+) and Br(-)-selective electrodes. All of the isotherm shapes in the mixture were classical. Activity coefficients were determined for the transfer between water and hydroorganic mixtures. Comparison of the adsorption isotherms obtained for different surfactant chemical potentials showed that at higher concentrations of free surfactant, the aggregation state does not depend on the nature of the surrounding media. A study of the influence of the ionic strength showed that there is a common intersection point in each hydroorganic mixture. This point corresponds to the charge compensation point and depends on the nature of the surrounding medium. PMID- 12885514 TI - Adsorption isotherms of homologous alkyldimethylbenzylammonium bromides on sodium montmorillonite. AB - The adsorption of homologous alkyldimethylbenzylammonium bromides, [C(6)H(5)CH(2)N(CH(3))(2)R]Br, on sodium montmorillonite from aqueous NaCl solutions at room temperature has been studied. R stands for the methyl-, butyl-, hexyl-, octyl-, decyl-, and dodecyl-group, and the corresponding ammonium cations will be denoted as C1+, C4+, C6+, C8+, C10+, and C12+, respectively. C1+, the reference cation, attains the plateau region of adsorption at a level close to the cation exchange capacity (CEC) of the clay. The chain-length dependence on adsorptivity of the homologous cations exhibits an unexpected peculiarity. In the case of short-chain homologues of C1+ their adsorption onto sodium montmorillonite decreases in the order C1+>C4+>C6+. This behavior is due, presumably, to the growing steric hindrances at the surface of clay, which occur because of the limited area available for the bulky organic cations at the exchange sites. These limitations appear to be out-balanced in the case of higher homologues for which the increasingly growing hydrophobic effects lead to the expected sequence of adsorptivity of the cations, i.e., C1+ 6.6. Adsorption of humic acid onto the chitosan-coated granules was found to be strongly pH-dependent. Significant amounts of humic acid were adsorbed under acidic and neutral pH conditions, but the adsorption capacity was reduced remarkably with increasing solution pH values. The adsorption isothermal data under various initial humic acid concentrations (at the same solution pH value) can be adequately modeled by the Langmuir and Freundlich models. X-ray photoelectron spectroscopy (XPS) revealed that the amino groups of the chitosan layer were protonated due to humic acid adsorption, suggesting the formation of organic complex between the protonated amino groups and humic acid. Kinetic study indicated that the adsorption process was transport-limited at low solution pH values, but became both transport- and attachment-limited at high solution pH values. PMID- 12885517 TI - Influence of copper content on NO removal of the activated carbon fibers produced by electroplating. AB - In this study, the activated carbon fibers (ACFs) on which copper metal was deposited by electroplating were used to remove nitric oxide (NO). N(2)/77 K adsorption isotherm characteristics, including the specific surface area and micropore volume, were investigated by BET and T-plot methods. NO removal efficiency was confirmed by gas chromatographic technique. From the experimental results, the copper content supported on ACFs led to an increase in the NO conversion, in spite of the decrease of the specific surface area or the micropore volume of ACFs. Consequently, the presence of Cu on ACFs played an important role in improving the NO reduction into O(2) and N(2), which was mainly attributed to the catalytic reactions of Cz-NO-Cu. PMID- 12885518 TI - New thermodynamically consistent competitive adsorption isotherm in RPLC. AB - A new equation of competitive isotherms was derived in the framework of the ideal adsorbed solution (IAS) that predicts multisolute adsorption isotherms from single-solute isotherms. The IAS theory makes this new isotherm thermodynamically consistent, whatever the saturation capacities of these single-component isotherms. On a Kromasil-C(18) column, with methanol-water (80/20 v/v) as the mobile phase, the best single-solute adsorption isotherm of both toluene and ethylbenzene is the liquid-solid extended multilayer BET isotherm. Despite a significant difference between the monolayer capacities of toluene (370 g/l) and ethylbenzene (170 g/l), the experimental adsorption data fit very well to single component isotherms exhibiting the same capacities (200 g/l). The new competitive model was used for the modeling of the elution band profiles of mixtures of the two compounds. Excellent agreement between experimental and calculated profiles was observed, suggesting that the behavior of the toluene-ethylbenzene adsorbed phase on the stationary phase is close to ideal. For example, the concentrations measured for the intermediate plateau obtained in frontal analysis differ by less than 2% from those predicted by the IAS model. PMID- 12885519 TI - Lithium adsorption by acid and sodium amberlite. AB - In this paper we used a previously reported model for examining the adsorption of nonelectrolytes in solution by solid adsorbents to study the adsorption of lithium(I) cations by acid and sodium amberlites, which is an ion-exchange process. Based on the results, both are equilibrium processes and obey a kinetic law with a unity partial order in the Li+ concentration. The kinetic results were used to calculate the specific rate constants and thermodynamic activation functions involved. Also, equilibrium isotherms were used to determine the corresponding ion-exchange capacities, the individual equilibrium constants, and the thermodynamic functions for the overall process. PMID- 12885520 TI - Surface protonation data of kaolinite-reevaluation based on dissolution experiments. AB - The aim of the present study is to compare available surface titration curves of kaolinite, to explain the differences between them, and to constrain their interpretation based on predictions of surface protonation that emerged from dissolution experiments. Comparison of six surface titration curves obtained at 25 degrees C reveals significant discrepancies, both in the shape of the curves and in the pH of the point of zero net proton charge (pH(PZNPC)). Based on an analysis of the different sites available for adsorption on kaolinite surfaces we conclude that different kaolinite samples are expected to have similar pH(PZNPC). Therefore, the major reason for the differences in the observed surface protonation is related to the different ways in which the pH(PZNPC) was determined. To compare the titration curves, some of the curves were recalculated so that the proton surface concentrations of all the titration curves would be zero around pH 5. As a result, we obtained a good agreement between the titration curves. A prediction of the molar fraction of protonated sites was retrieved from modeling of kaolinite dissolution reaction and was compared to the protonation data obtained from surface titration. The model successfully predicts the surface protonation data of most of the surface titrations. PMID- 12885521 TI - Myelin growth and initial dynamics. AB - During the dissolution of solid surfactants in water, various types of nonequilibrium microstructures have been observed. The most important one is the myelin growth which can be observed when some poorly water soluble surfactants such as phosphatidylcholine (PC), Aerosol-OT (AOT), etc. are contacted with water. In this study initial myelin growth for a period of 2-4 s is studied both qualitatively as well as quantitatively in all the directions with respect to water flow in a PC system using digital video microscopy. Further, overall myelin growth is studied by means of optical microscopy to understand the effect of distance between cover slip and glass slide on myelin growth. Experiments are also performed to study effect of additives (silica) to lamellar phase on diffusion coefficients. It has been observed that the presence of silica particles causes extensive coiling of myelin structures. The mechanism of water transport into the lamellar phase during myelin growth is investigated by using silica in a colloidal range as dopant material. PMID- 12885522 TI - Interaction of proteins with weak amphoteric charged membrane surfaces: effect of pH. AB - Weak amphoteric charged membranes were prepared by the graft copolymerization of poly(ethylene glycol) (PEG) derivatives with pendant ionizable groups onto polyethylene (PE) porous membranes. Two types of weak amphoteric charged membranes and two types of weak single charged membranes were prepared. The pH dependence of the protein (fluorescein isothiocyanate-labeled bovine serum albumin, FITC-BSA) adsorption onto the membranes was investigated by fluorescence spectroscopy. The interfacial charge properties of the membranes and protein were also characterized at different pH values by streaming potential and electrophoretic light scattering (ELS) measurements, respectively. The adsorbed amount onto each ionic PEG chain grafted membrane showed a uniform maximum value near the isoelectric point (IEP) of the protein (pH 4.1). On both sides of the IEP (pHs 3.3 and 7.2), the adsorption experiments and zeta (zeta) potential measurements were well correlated: the contribution of electrostatic interaction was dominant for the protein adsorption behavior. In the alkaline condition (pH 10.2), the adsorption experiments contradict the zeta potential measurements. It suggested that the conformational change of protein molecule influenced the adsorption behavior. Finally, these results indicated the potential of controlling the protein-ionic PEG chain interaction on the membrane surfaces by the pH adjustment of the outer solution. PMID- 12885523 TI - Anomalous nanostructured titanium dioxide. AB - Titanium dioxide nanoparticles prepared in water-in-oil microemulsion droplets by controlled hydrolysis of TiCl(4)-generated crystalline nanoparticles of sizes from 115 nm down to 6 nm diameter depending on the size of the aqueous core of the micellar droplets. Powder X-ray diffraction of the vacuum-dried product (without sintering) indicated the presence of an unusual type of orthorhombic crystal structure nearly similar to titanium dioxide crystals prepared at high pressure. On gradual heating up to 900 degrees C these metastable crystals are converted into relatively more stable nanorods perhaps through making and breaking of the Ti-O-Ti bonds. It has been concluded that chemical pressure generated within the constrained volume of aqueous core of the reverse micellar droplets is responsible for the unusual crystal structure of TiO(2) nanoparticles. PMID- 12885524 TI - Characterization of silica-coated hematite and application to the formation of composite particles including egg yolk PC liposomes. AB - According to the method of Ohmori et al. (J. Colloid Interface Sci. 150 (1992) 594), a procedure is examined for the buildup of uniform silica layers on monodispersed hematite particles. It appears that the silica layer resulting is homogeneous and the layer thickness is controlled by the concentration of tetraethylorthosilicate (TEOS) in the medium. Further, egg PC liposomes, a typical biocolloid, are introduced onto the silica-coated hematite particle. The formation was proceeded by two types of processes: (1) heterocoagulation between the silica-coated hematite and egg PC liposomes by controlling the concentration of LaCl(3) in the medium, or (2) buildup using two proteins (lysozyme or cytochrome C) as binder molecules. These results were analyzed by zeta-potential measurements and a contact-type X-ray microscope, which is a unique technique for obtaining X-ray images of biological specimens in water with high resolution. PMID- 12885525 TI - Charge and salt-driven reentrant order-disorder and gas-solid transitions in charged colloids. AB - Monte Carlo simulations have been performed for aqueous charged colloidal suspensions as a function of effective charge density (sigma) on the particles and salt concentration C(s). We vary the effective charge density in our simulations over a range where a reentrant solid-liquid transition in suspensions of silica and polymer latex particles has been reported by Yamanaka et al. (Phys. Rev. Lett. 80 (1998) 5806). We show that at low ionic strengths a homogeneous liquid-like ordered suspension undergoes crystallization upon increasing sigma. Further increase in sigma resulted once again in a disordered state, which is in agreement with experimental observations. In addition to this reentrant order disorder transition, we observe an inhomogeneous-to-homogeneous transition in our simulations when salt is added to the disordered inhomogeneous state. This inhomogeneous-to-homogeneous disordered transition is analogous to the solid-gas transition of atomic systems and has not yet been observed in charged colloids. The reported experimental observations on charged colloidal suspensions are discussed in the light of present simulation results. PMID- 12885526 TI - Electroactive gold nanoparticles protected by 4-ferrocene thiophenol monolayer. AB - Numerous reports have focused on ferrocene-terminated electroactive self assembled monolayers (SAMs) on a flat Au surface but only a few on ferrocene SAMs on Au colloid. In this paper, we employ 4-ferrocene thiophenol as a novel capping agent to produce electroactive gold nanoparticles in consideration of the peculiar pi-conjugated structure. Transmission electron microscopy shows the narrow-dispersed gold core with an average core diameter of ca. 2.5 nm. UV/vis spectra examine the pi-conjugated structure of 4-ferrocene thiophenol and surface plasmon absorbance of the indicated gold nanoparticles. X-ray photoelectron spectroscopy reveals electronic properties of the Au core and thiol ligands. Electrochemical measurement shows that the oxidation peak current is proportional to the scan rate, indicating the electrode process is controlled by adsorbed layer reaction. The formal potential of the Fc-MPCs is compared with that of free ferrocene in MeCN solution and the Fc-SAMs. The shifts are attributed to the phenyl moiety in the 4-ferrocene thiophenol and dielectric constant of the solvation environment. PMID- 12885527 TI - CO adsorption on copper in alkaline electrolytes: an electrochemical and ellipsometric study. AB - The influence of CO on the passive behavior of copper was analyzed in the potential region near the rest potential in borax solutions (pH 9.2) by cyclic voltammetry, ellipsometry, and surface charge determination techniques. The oxide formation is explained as a sequence of Cu(2)O growth, cation adsorption, Cu(II), and dissolution steps, similarly to previously reported investigations for the metal in free CO solutions. The CO adsorption hinders the cationic defect in the outer oxide layer and accelerates the Cu(2)O growth both at open circuit and in controlled potential experiments. The isoelectric point, iep, obtained at pH 10.8 for both the metal Cu and CuO particles in KCl solutions shifts to pH 10.1 for copper particles in the presence of CO. The iep indicates a CuO coating on Cu metallic particles in the absence of CO. PMID- 12885528 TI - Electrophoresis of a charge-regulated spheroid along the axis of an uncharged cylindrical pore. AB - Boundary effects can have a profound influence on the electrophoretic behavior of a charged entity, in particular, when the entity is nonspherical and its surface conditions are dependent upon the nearby environment. In this study, the electrophoresis of a spheroid along the axis of an uncharged cylindrical pore is analyzed for the case where the electrical potential is low and the applied electric field is weak. We consider the case where the surface of a particle contains dissociable acidic and basic functional groups, which simulate biological colloids and entities covered by an artificial membrane. This leads to a mixed-type boundary value problem, which extends the conventional constant surface-potential and constant-surface-charge-density models to a more general case. The effects of the particle aspect ratio, the relative magnitudes of particle and pore, the thickness of the double layer surrounding a particle, and the pH of the liquid phase on the electrophoretic mobility of a particle are investigated. Several interesting results are observed; for example, if the volume of a particle is fixed, its mobility may have a local maximum as the relative magnitudes of its two axes vary. PMID- 12885529 TI - Studies of Athabasca asphaltene Langmuir films at air-water interface. AB - Asphaltenes are present in heavy oils and bitumen. They are a mixture of hydrocarbons having complex structures of polyaromatic rings and short side chains. In general, the high-molecular-weight asphaltene is the most aromatic fraction with the highest number of side chains and the low-molecular-weight asphaltene contains the lowest number of side chains, while the number of side chains of the whole asphaltene fraction lies in between. In this study, asphaltenes were extracted and/or fractionated from Athabasca oil sand bitumen. Subfractions of high and low molecular weight and the whole asphaltenes were characterized using a Langmuir trough and complementary techniques such as VPO, FTIR, AFM, and contact angle measurements. At an air-water interface, amphiphilic asphaltene molecules can form a monolayer. Various fractions (high, low, and whole) of the asphaltene molecules behave similarly at the air-water interface, characterized by close resemblance of their surface pressure-area, hysteresis, and relaxation isotherms. The high-molecular-weight asphaltene is the most expanded fraction, while the low-molecular-weight asphaltene fraction is the most condensed, with the whole asphaltene lying in between. At the air-water interface a monolayer of the low-molecular-weight asphaltene relaxes at a faster rate than one of the high-molecular-weight asphaltene. PMID- 12885530 TI - Estimation of magnetic particle clustering in magnetic fluids from static magnetization experiments. AB - The volume fraction dependence of the static magnetization of two magnetic fluids with different degrees of steric stabilization was measured at low field values (0-10 kA/m) and it was found to be nonlinear for both magnetic fluids. The nonlinearity is more pronounced in the case of the less stabilized magnetic fluid. The experimental data were processed by nonlinear regression using an analytical model for the formation of chain-like magnetic particle aggregates in magnetic fluids. The calculated dependence on the degree of steric stabilization, magnetic field, and sample concentration of the mean number of particles per chain was in the range (1-1.04). PMID- 12885531 TI - Studies on the sea surface microlayer. II. The layer of sudden change of physical and chemical properties. AB - Seawater samples of the sea surface microlayer were obtained from the ocean, coastal waters, and laboratory imitation experiments adopting glass plate, rotating drum, screen, and funnel samplers. The result was that surface microlayer samples of thickness 50 microm could be taken, not by the popular screen and funnel techniques, but by glass plate or rotating drum techniques. The layer of sudden change of physical and chemical properties in the surface microlayer was found at 50 microm below the sea-air interface. These physical and chemical properties included tens of physical and chemical parameters, such as concentrations of organic matter, nutrients, and dissolved trace metals, biochemical oxygen demand, chemical oxygen demand chlorophyll-a, surface tensions, and pH. Therefore, it was suggested that the layer of sudden change of physical and chemical properties in surface seawater should serve as a basis for defining the sea surface microlayer should, and be regarded as the practical operational thickness of the sea surface microlayer. The apparent sampling thickness of the sea surface microlayer from surface seawater should be 50+/-10 microm. PMID- 12885532 TI - Surfactant driven flows overlying a hydrophobic epithelium: film rupture in the presence of slip. AB - Both for tear films and along the airways within the lung, one has an extremely thin fluid layer overlying a biological substrate; in both cases surfactants either of natural origin, or artificially introduced, are important in driving fluid flows. There is evidence that slip can occur when hydrophilic liquids, similar to mucus or aqueous tear films, overlie hydrophobic epithelium. Utilizing results from recent experimental findings we examine the possible influence of slip upon tear film rupture, important in so-called dry eye, and upon surfactant induced flows within the lung, used in surfactant replacement therapy. PMID- 12885533 TI - Self-assembled multilayer films based on a Keggin-type polyoxometalate and polyaniline. AB - Multilayer ultrathin films were fabricated from partially doped polyaniline (PAN) and a Keggin-type polyoxometalate [alpha-SiW(12)O(40)](4-) (alpha-SiW(12)) in aqueous solution via the layer-by-layer self-assembly technique and characterized by UV-vis, FTIR, and X-ray photoelectron spectra (XPS), ellipsometry, scanning electron microscopy (SEM), and atomic force microscopy (AFM). UV-vis spectroscopy shows that the absorbance values at characteristic wavelengths of the multilayer films increase almost linearly with the number of PAN/alpha-SiW(12) bilayers, suggesting that the deposition process is regular and highly reproducible from layer to layer. FTIR and XPS spectra confirm the incorporation of alpha-SiW(12) and PAN into the films. Ellipsometric measurements show that the PAN/alpha SiW(12) bilayer thickness increases with the increasing PAN solution concentration. SEM and AFM images indicate that the film surface is relatively uniform and smooth. In addition, the electrical conductivities of the multilayer films doped with hydrochloric acid were also measured. PMID- 12885534 TI - Characterizing flocculation under heterogeneous turbulence. AB - This study investigated the impact of turbulent heterogeneity in a flocculation reactor on particle aggregation and breakup. In particular, the influence of average characteristic velocity gradient (G), particle concentration, and coagulation mechanism (sweep floc vs charge neutralization) on the floc growth, steady-state size, and variance was analyzed. Experiments were performed in a bench-scale reactor with a low-shear axial-flow impeller using a photometric dispersion analyzer (PDA). Results indicated that as G increased, floc growth increased while the mean size and variance in the floc size distribution decreased. In addition, floc growth, mean size, and variance increased with increasing primary particle concentration and when the coagulation mechanism was switched from charge neutralization to sweep floc. Lastly, floc growth, mean size, and variance were found to vary spatially in the reactor at low G values with larger floc size and growth rate in the bulk region and a larger variance in the impeller discharge region. PMID- 12885535 TI - On the behavior of electrokinetic streaming potential during protein filtration with fully and partially retentive nanopores. AB - An experimental investigation of the electrokinetic streaming potentials of both fully and partially retentive nanopores as compared with the filtration progress of dilute globular protein solution under different surface charge conditions was performed using hollow fibers. The streaming potential is generated by the electrokinetic flow effect within the electric double layer of the charged surface. Depending on the solution pH, both the protein and the pore wall can be either repulsive or attractive due to the long-range electrostatic interaction. The repulsive electrostatic interaction allows the protein particles to stay in a suspended state above the outer surface of hollow fibers instead of being deposited. The apparent streaming potential value at partially retentive pores is larger than that at fully retentive pores for the oppositely charged case; however, the opposite behavior is shown for the same-charged case. The axial position-dependent streaming potential was also observed in order to explore the development of a concentration polarization layer during the cross-flow filtration. The time evolution of the streaming potential during the filtration of protein particles is related to the filtrate flux, from which it can be found to provide useful real-time information on particle deposition onto the outer surfaces of hollow fibers. PMID- 12885536 TI - Exfoliation and immobilization of anionic iron porphyrin in layered double hydroxides. AB - Mg-Al layered double hydroxide (LDH) intercalated with glycinate anions was synthesized through co-precipitation and exfoliated in formamide and the single layer suspension was reacted with an aqueous iron porphyrin (FeTDFSPP) solution. The new composite was characterized by powder X-ray diffraction, UV-vis, FTIR, and electron paramagnetic spectroscopies and thermal analyses (simultaneous TG/DSC). The analyses demonstrated that glycinate anions continue to be intercalated and the anionic iron porphyrin is adsorbed at the surface of the layered double hydroxide crystals. PMID- 12885537 TI - Oxidation of cyclohexane catalyzed by metal-ion-exchanged zeolites. AB - The ion-exchange rates and capacities of the zeolite NaY for the Cu(II), Co(II), and Pb(II) metal ions were investigated. Ion-exchange equilibria were achieved in approximately 72 h for all the metal ions. The maximum ion exchange of metal ions into the zeolite was found to be 120 mg Pb(II), 110 mg Cu(II), and 100 mg Co(II) per gram of zeolite NaY. It is observed that the exchange capacity of a zeolite varies with the exchanged metal ion and the amount of metal ions exchanged into zeolite decreases in the sequence Pb(II) > Cu(II) > Co(II). Application of the metal-ion-exchanged zeolites in oxidation of cyclohexane in liquid phase with visible light was examined and it is observed that the order of reactivity of the zeolites for the conversion of cyclohexane to cyclohexanone and cyclohexanol is CuY > CoY > PbY. It is found that conversion increases by increase of the empty active sites of a zeolite and the formation of cyclohexanol is favored initially, but the cyclohexanol is subsequently converted to cyclohexanone. PMID- 12885538 TI - Interfacial deformation between an impacting water drop and a silicone-oil surface. AB - Detailed observations have been conducted on the interfacial deformation of a silicone oil surface and a water drop falling on it. Eleven kinds of silicone oils with wide variations of kinematic viscosity, nu(T)=1-10(5) mm(2)/s, have been tested. The oil surface is disturbed by a water drop with a diameter d(L)=3.1 mm, which falls freely on it from a height of 100-1000 mm. Special attention has been directed to the maximum depth of the cavity formed on the oil surface D(M) and to the maximum diameter of the water drop spreading on the oil surface d(M). We have categorized the configurations of the oil cavity into seven patterns, and those of the water drop at the oil-water interface into five patterns. The maximum cavity depth D(M)/d(L) can be well correlated by a dimensionless group Re(TL)We(TL), where Re(TL) is Reynolds number based on d(L) and nu(T) and We(TL) is Weber number with the water density and surface tension of oil. The maximum diameter of the impacting water drop d(M)/d(L) can be correlated by the Reynolds number with a viscosity of water (Re(L)) and the Ohnesorge number (Oh). Moreover, the condition under which the impacting water drop is smashed into pieces has been also examined based on Re(L) and Oh. PMID- 12885539 TI - Interfacial effects in the spreading kinetics of liquid droplets on solid substrates. AB - Several theories deal with the spreading kinetics of liquids on solid substrate, most of which relate the rate of spreading to the surface tension and the viscosity of the liquid. Measurements of the spreading of a number of liquids exhibiting a wide range of surface tension and viscosity on dry soda-lime glass have been carried out to validate the proposed models. The measurements used a small droplet of constant volume to minimize gravitational effects. The contact radius was acquired as a function of time by an image analysis system. It was noted that power law theories describe the spreading rate for silicone oil on glass. However, significant departures were noted in the case of other liquids. Mechanistic considerations of our data suggest that equal volume droplets of similar surface tension and of diverse viscosity spread to the same area but at different rates. On the other hand, the spreading rate of glycerine, which exhibits incomplete spreading on glass, and that of silicone oil, with comparable viscosity behave similarly. These observations seemingly support the view that surface tension acts to retain the spherical shape of the droplet, whereas the difference between the solid-liquid and solid-vapor interfacial energies acts to enlarge the contact area. In the meantime, viscous dissipation acts to retard the spreading rate, past a constant rate regime. PMID- 12885540 TI - First-order curvature corrections to the surface tension of multicomponent systems. AB - The dependence of surface tension on curvature is investigated for the case of an equilibrium phase coexistence in multicomponent systems. Employing Gibbs's method of description of heterogeneous systems, an equation is derived to determine the dependence of surface tension on curvature for widely arbitrary paths of variation of the independent thermodynamic parameters. It is supposed hereby merely that the temperature is kept constant and that the variations of the different molar fractions are such that the radius of the dividing surface varies monotonically in dependence on the change of the state parameters of the ambient phase along any of the chosen paths. In the analysis, an approach developed by Blokhuis and Bedeaux for one-component systems is utilized. It relies on the expansion of the surface free energy on curvature of the dividing surface. An equation is derived that connects the first-order correction term in the expansion with the interaction potential of the particles in the multicomponent solution and with the two-particle distribution functions in the planar interfacial layer between the two phases coexisting in equilibrium at planar interfaces. The connection of the first-order curvature correction to the surface tension and the first moment of the pressure tensor at a planar interface is analyzed as well. PMID- 12885541 TI - A mathematical model for an expanding foam. AB - A theoretical and numerical model is presented for the shape evolution of the thin liquid films separating the gas bubbles in a foam. The motion is due to capillary action, surface tension gradients, and the overall expansion of the foam. The expansion is the result of the increase in gas content with time. Process modeling is accomplished via the solution of three coupled partial differential equations. Two time scales are included in the model: a process time and a drying or curing time. It is demonstrated that the amount of surfactant is the dominant control mechanism for the final film thickness. If sufficient surfactant is present, the films will be shown to dilate uniformly in space. A number of known features of expanding foams are reproduced by the model. PMID- 12885542 TI - Phase behavior in the system water-butylammonium decanoate-decane. AB - The phase diagram at 298.2 and 281.2 K for the ternary system water-butylammonium decanoate-decane has been determined. An isotropic solution phase extending from the water corner to the decane corner was obtained without any macroscopic phase separation. A large two-phase region extending from the water-decane axis was also observed. In addition to these phases, an anisotropic and optically birefrigent liquid crystalline phase is formed below 290.3 K. Sound velocity, conductivity, and viscosity measurements were used successfully for studies of microstructural transitions in the isotropic solution phase. From these measurements, it was concluded that a transition from normal micelles to reverse micelles takes place while passing through a bicontinuous region. PMID- 12885543 TI - Predictive network modeling of single-phase non-Newtonian flow in porous media. AB - The majority of complex fluids used in oil-field applications are polymeric solutions exhibiting shear-thinning (pseudoplastic) behavior in solution. We study the flow of power-law fluids in porous media using network modeling. We use realistic representations of porous media that capture the geometry and topology of sands and sandstones as input to our flow model. The bulk rheology (variation of viscosity with shear rate) is used to define a relationship between pressure drop and average flow velocity in each pore. We then compute the variation of average velocity with apparent viscosity for the whole network. We successfully predict the results of four experiments from different sources in the literature. The predictions are superior to those using simplified capillary bundle models and involve no adjustable parameters, but are limited to polymer exhibiting simple shear-thinning behavior and require a detailed description of the pore space of interest. PMID- 12885544 TI - Measuring the swelling behavior of polymer microspheres with different cross linking densities and the medium-dependent color changes of the resulting latex crystal films. AB - We measured the size changes of cross-linked polymer microspheres of narrow size distribution with different cross-linking densities by the in situ swelling method. The swelling behavior of these cross-linked polymer microspheres revealed that their maximal swelling degree linearly decreased as a function of the cross linker content from 6.65 at 1.36 mol% to 2.43 at 3.25 mol%. By means of UV-vis spectra, we observed the spectral changes of the latex crystal films composed of these cross-linked polymer microspheres when we filled the interstices of the latex crystal films with different fluid media. Some succeeding peak shifts were observed, which may have resulted from the movement or the swelling of the polymer microspheres in the latex crystals. PMID- 12885545 TI - Characterization of charge neutralization reaction in aqueous solution using the membrane separation method. AB - Colloid titration and streaming current detector are two popular techniques used to determine the charge density of dissolved and colloidally dispersed polyelectrolytes. However, both techniques are based on some assumptions that may not be valid under some circumstances. In this paper, a membrane separation technique developed to determine the charge density and characterize a charge neutralization reaction is described. The technique characterizes the reaction between cationic and anionic charge carriers by measuring the counterions released during reaction or the unreacted small molecule charge carriers. Membranes with selected cutoff pore size were used to separate the species of interest from the bulk of the reaction mixture. Three different charge determination techniques were also compared. PMID- 12885546 TI - Reconformation of polyvinylamine adsorbed on glass fibers. AB - Surface area exclusion chromatography was used to investigate the reconformation of fully hydrolyzed polyvinylamine. The polymer is adsorbed on stacked glass fiber filters constituting the stationary phase while the polymer solution is injected at the inlet of the chromatography column. From numerical simulation and experimental chromatograms of nonreconforming polyelectrolytes, the amount of polymer adsorbed per filter represented as a function of the filter position along the column (the histogram) was determined to be continuously decreasing and not to depend on the rate of elution. For polyvinylamine, the histograms are peaked and the height of the peak was determined to depend greatly on the rate of polymer supply to the column that was controlled by monitoring the polymer concentration and/or the rate of elution (mass-transfer-controlled adsorption). Modifications in the adsorption on the successive filters were converted into changes in the interfacial area of adsorbed molecules taking into account model histograms as well as experimental adsorption histograms of non reconforming systems. Macromolecule concentration in the mobile phase and contact time between solute and adsorbed molecules were determined to be the two parameters controlling the extent of polymer desorption. The unusual shape of the histogram thus was attributed to reconformation of the adsorbed polymer, which was stimulated by interfacial exchange between segments belonging to trains of adsorbed macromolecules and chain segments of solute ones. PMID- 12885547 TI - Surface properties of poly(N-monoalkylmaleamic acid-alt-styrene) sodium salts: effect of the molecular weight and the side chain length. AB - The surface properties of poly(N-monoalkylmaleamic acid-alt-styrene) sodium salts are studied as a function of the molecular weight and the size of the linear alkyl lateral chain of the polyelectrolyte. The experimental results are well described by the Gibbs-Szyszkowski treatment. Both the surface tension behavior and the standard free energy of adsorption depend on the polyelectrolyte side chain and on the average molecular weight, M(w). An M(w)-dependent contribution to the free energy of adsorption ranging from -1.21 to -1.05 kJ for mole of methylene groups is found. The area covered by monomer units increases with M(w) and the sizes of side chains are similar to those reported in small-molecule systems. The nature of the functional group amide in the side chain has practically no effect on the surface properties as compared with the ester group in this kind of polyelectrolytes. PMID- 12885548 TI - Photoinduced interaction between fluorescein ester derivatives and CdS colloid. AB - The photoinduced interaction of fluorescein ester derivatives, fluorescein's butyl ester (FL4) and fluorescein's anthraquinone-methyl ester (FL-AQ), and colloidal CdS was examined by absorption, fluorescence spectroscopy, and photoinduced ESR spectroscopy. It is found that FL4 and FL-AQ molecules are adsorbed on the surfaces of CdS colloid by an electrostatic interaction forming the surface complex of the type CdS-FL4 or CdS-FL-AQ. The apparent association constant (K(app)) and the degree of association (alpha) of CdS-FL4 obtained from absorption spectra are 2.25 x 10(4) M(-1) and 0.78, respectively. The values of K(app) and alpha of CdS-FL4 as determined from fluorescence spectra are 1.54 x 10(4) M(-1) and 0.82, respectively, which matches well with that determined from the absorption spectra changes. And the values of K(app) and alpha of CdS-FL-AQ obtained from absorption spectra are 4.18 x 10(4) M(-1) and 0.83, respectively. These data indicate that there is a strong interaction between the dye and the CdS particle surface. But there was no evidence for interfacial electron transfer from FL4 or FL-AQ to colloidal CdS by photoinduced ESR experiments. The fluorescence quenching is due to the formation of a nonfluorescent complex. The related phenomena are discussed in this paper. PMID- 12885549 TI - X-ray photoelectron spectroscopic studies of hydrophilic surfaces modified via admicellar polymerization. AB - Admicellar polymerization with styrene monomer was used to coat the surface of two porous solids: titanium dioxide and alumina. X-ray photoelectron spectroscopy (XPS) measurements clearly indicate that after admicellar polymerization, organic material and surfactant are present on the surface of the solids. Water washes, performed immediately after admicellar polymerization, were successful in removing approximately 30% of the organic material, presumably mostly surfactant. The water wash was followed by Soxhlet extraction with toluene, with no measurable removal of organic material according to ignition loss measurements of the solid, gel permeation chromatography of the extracting solvent and also UV/VIS spectroscopy of the extracting solvent. However, there was a 5-15% drop in organic material after Soxhlet extraction according to XPS measurements. This difference is attributed to the difference in sampling; XPS samples only the exterior surface area while these bulk measurements sample both the interior and exterior surface area. This study details the ability of different washing steps to remove materials from the porous substrates used in this study. PMID- 12885550 TI - Growth and reactivity of silver clusters in amino polycarboxylic acid solutions. AB - Reduction and subsequent aggregation of silver ions in the presence of various chelating agents such as trans-1,2-diaminocyclohexanetetraacetic acid (DCTA) 1,2 propylenediaminetetraacetic acid (PDTA), diethylenetriaminepentaacetic acid (DTPA), and triethylenetetraaminehexaacetic acid (TTHA) have been studied by pulse radiolysis. Rate constants of formation and transient absorption spectra for the ligand-complexed Ag(0) and Ag(2)(+) have been determined. The redox potential of Ag(+)(L)/Ag(0)(L) becomes more negative than that of Ag(+)/Ag(0). Growth and reactivity of silver clusters were also studied in the presence of methyl viologen (MV(2+)). The kinetics of formation of the MV(+*) radical, produced by the same pulse as in the case of silver atoms, confirms the catalytic electron transfer toward super critical silver clusters in the presence of ligands. The results suggest that catalytic electron transfer activity of silver clusters depends on the ligand. PMID- 12885551 TI - Signaling systems, guided cell migration, and organogenesis: insights from genetic studies in Drosophila. AB - During development, cells change their position extensively. Although the basic cellular mechanisms involved in cell locomotion have been studied mostly in cultured cells, genetic and molecular approaches using model organisms are starting to shed light on the complex events influencing cell migration during development. Recent technical advances in following and analyzing migrating cells inside the living embryo offer the possibility of understanding how different signaling systems regulate the fundamental cellular processes underlying guided cell migration in vivo. In Drosophila melanogaster, studies of migrating cells have concentrated mainly on hemocytes, germ cells, border cells, and tracheal cells. Interestingly, most of these cells were recently shown to make different cellular extensions and to use receptor tyrosine kinases to sense the chemoattractive signal. This review describes our current understanding of how different signaling networks control guided migration in these four systems and discusses the impact of novel imaging techniques on the study of guided cell migration during development. PMID- 12885552 TI - Reciprocal interactions between olfactory receptor axons and olfactory nerve glia cultured from the developing moth Manduca sexta. AB - In olfactory systems, neuron-glia interactions have been implicated in the growth and guidance of olfactory receptor axons. In the moth Manduca sexta, developing olfactory receptor axons encounter several types of glia as they grow into the brain. Antennal nerve glia are born in the periphery and enwrap bundles of olfactory receptor axons in the antennal nerve. Although their peripheral origin and relationship with axon bundles suggest that they share features with mammalian olfactory ensheathing cells, the developmental roles of antennal nerve glia remain elusive. When cocultured with antennal nerve glial cells, olfactory receptor growth cones readily advance along glial processes without displaying prolonged changes in morphology. In turn, olfactory receptor axons induce antennal nerve glial cells to form multicellular arrays through proliferation and process extension. In contrast to antennal nerve glia, centrally derived glial cells from the axon sorting zone and antennal lobe never form arrays in vitro, and growth-cone glial-cell encounters with these cells halt axon elongation and cause permanent elaborations in growth cone morphology. We propose that antennal nerve glia play roles similar to olfactory ensheathing cells in supporting axon elongation, yet differ in their capacity to influence axon guidance, sorting, and targeting, roles that could be played by central olfactory glia in Manduca. PMID- 12885553 TI - A role for programmed cell death during early neurogenesis in xenopus. AB - In vertebrates, little is known on the role of programmed cell death (PCD) occurring within the population of dividing neural precursors and newly formed neuroblasts during early neural development. During primary neurogenesis, PCD takes place within the neuroectoderm of Xenopus embryos in a reproducible stereotypic pattern, suggesting a role for PCD during the early development of the CNS. We find that the spatio-temporal pattern of PCD is unaffected in embryos in which cell proliferation has been blocked and whose neuroecotoderm contains half the normal number of cells. This shows that PCD is not dependent on cell division. It further suggests that PCD does not solely function to regulate absolute cell numbers within the neuroectoderm. We demonstrate that PCD can be reproducibly inhibited in vivo during primary neurogenesis by the overexpression of human Bcl-2. Following PCD inhibition, normal neurogenesis is disrupted, as seen by the expansion of the expression domains of XSox-2, XZicr-2, XNgnr-1, XMyT 1, and N-Tubulin, XNgnr-1 being the most affected. PCD inhibition, however, did not affect the outcome of lateral inhibition. We propose, then, that PCD regulates primary neurogenesis at the level of neuronal determination. PMID- 12885554 TI - Role of Phox2b and Mash1 in the generation of the vestibular efferent nucleus. AB - The inner ear (vestibular and cochlear) efferent neurons are a group of atypical motor-like hindbrain neurons which innervate inner ear hair cells and their sensory afferents. They are born in the fourth rhombomere, in close association with facial branchial motor neurons, from which they subsequently part through a specific migration route. Here, we demonstrate that the inner ear efferents depend on Phox2b for their differentiation, behaving in that respect like hindbrain visceral and branchial motor neurons. We also show that the vestibular efferent nucleus is no longer present at its usual site in mice inactivated for the bHLH transcription factor Mash 1. The concomitant appearance of an ectopic branchial-like nucleus at the location where both inner ear efferents and facial branchial motor neurons are born suggests that Mash1 is required for the migration of a subpopulation of rhombomere 4-derived efferents. PMID- 12885555 TI - The ventralized ogon mutant phenotype is caused by a mutation in the zebrafish homologue of Sizzled, a secreted Frizzled-related protein. AB - The BMP signaling pathway plays a key role during dorsoventral pattern formation of vertebrate embryos. In zebrafish, all cloned mutants affecting this process are deficient in members of the BMP pathway. In a search for factors differentially expressed in swirl/bmp2b mutants compared with wild type, we isolated zebrafish Sizzled, a member of the secreted Frizzled-related protein family and putative Wnt inhibitor. The knockdown of sizzled using antisense morpholino phenocopied the ventralized mutant ogon (formerly also known as mercedes and short tail). By sequencing and rescue experiments, we demonstrate that ogon encodes sizzled. Overexpression of sizzled, resulting in strongly dorsalized phenotypes, and the expression domains of sizzled in wild type embryos, localized in the ventral side during gastrulation and restricted to the posterior end during segmentation stages, correlate with its role in dorsoventral patterning. The expanded expression domain of sizzled in ogon and chordino together with its downregulation in swirl suggests a BMP2b-dependent negative autoregulation of sizzled. Indicating a novel role for a secreted Frizzled related protein, we show that, in addition to the BMP pathway, a component of the Wnt signaling pathway is required for dorsoventral pattern formation in zebrafish. PMID- 12885556 TI - A novel inducible element, activated by contact with Rathke's pouch, is present in the regulatory region of the Rpx/Hesx1 homeobox gene. AB - Reciprocal inductive interactions are postulated to play a role in the determination and differentiation of the pituitary gland and the ventral hypothalamus. The homeobox gene Rpx/Hesxl is expressed during gastrulation in the anterior endoderm, prechordal plate, and the prospective cephalic neural plate, and at later stages of development in Rathke's pouch, the primordium of the pituitary. We have defined the regulatory elements necessary for proper spatial and temporal expression during development in transgenic mice using lacZ reporter genes. Proper spatial and temporal expression in the anterior endoderm prechordal plate and anterior neural plate can be recapitulated with as little as 568 bp of upstream sequence and intragenic sequence containing the first exon and intron. Late-stage expression in Rathke's pouch requires additional negative and positive regulatory elements. Interestingly, deletion analysis uncovered an element that directs transgene expression to a region of the hypothalamus that lies in direct contact with Rathke's pouch. In vitro tissue recombination experiments have established that this expression is induced by contact with the pouch. We propose that this element may be present in other genes that normally respond to signals emanating from the pouch during the development of the hypothalamic-pituitary axis. The Rpx-lacZ transgenic mice provide a novel model system for the molecular dissection of inductive cell signaling during pituitary development. PMID- 12885557 TI - Sox10 is required for the early development of the prospective neural crest in Xenopus embryos. AB - The Sox family of transcription factors has been implicated in the development of different tissues during embryogenesis. Several mutations in humans, mice, and zebrafish have shown that depletion of Sox10 activity produces defects in the development of neural crest derivatives, such as melanocytes, ganglia of the peripheral nervous system, and some specific cell types as glia. We have isolated the Xenopus homologue of the Sox10 gene. It is expressed in prospective neural crest and otic placode regions from the earliest stages of neural crest specification and in migrating cranial and trunk neural crest cells. Loss-of function experiments using morpholino antisense oligos against Sox10 produce a loss of neural crest precursors and an enlargement of the surrounding neural plate and epidermis. This effect of Sox10 depletion is produced during some of the earliest steps of neural crest specification, as is shown by the inhibition in the expression of Slug and FoxD3, which are early markers of neural crest specification. In addition, we show that Sox10 depletion leads to an increase in apoptosis and a decrease in cell proliferation in the neural folds, suggesting that Sox10 could work as a survival as well as a specification factor in neural crest precursors during premigratory stages. Although some of the deficiencies found in the Waardenburg syndrome and in the Hirschprung disease could be associated with a failure of the development of crest derivatives during the late phase of its development, or even during adulthood, our results suggest that inhibition of Sox10 activity produces an earlier failure of neural crest precursors. In experiments where melanocytes and ganglia were induced in vivo and in vitro, we were able to block their development by inhibiting Sox10 activity. These results are compatible with an additional late role of Sox10 on development of neural crest derivatives, as it has been previously proposed. We show that Sox10 expression is dependent on FGF and Wnt activity, both in the neural crest and in the otic placode territories. Finally, in order to establish the position of Sox10 in the hierarchical cascade of gene activation required for neural crest specification, we used inducible forms of the wild type and dominant negatives for the Snail and Slug genes. Our results show that Snail is able to control Sox10 expression. However, the overexpression of Slug was not able to upregulate Sox10 expression. Taken together, these results indicate that Sox10 may lie between Snail and Slug in the genetic cascade that controls neural crest development. PMID- 12885558 TI - The expression of the proximodistal axis patterning genes Distal-less and dachshund in the appendages of Glomeris marginata (Myriapoda: Diplopoda) suggests a special role of these genes in patterning the head appendages. AB - The genes Distal-less, dachshund, extradenticle, and homothorax have been shown in Drosophila to be among the earliest genes that define positional values along the proximal-distal (PD) axis of the developing legs. In order to study PD axis formation in the appendages of the pill millipede Glomeris marginata, we have isolated homologues of these four genes and have studied their expression patterns. In the trunk legs, there are several differences to Drosophila, but the patterns are nevertheless compatible with a conserved role in defining positional values along the PD axis. However, their role in the head appendages is apparently more complex. Distal-less in the mandible and maxilla is expressed in the forming sensory organs and, thus, does not seem to be involved in PD axis patterning. We could not identify in the mouthparts components that are homologous to the distal parts of the trunk legs and antennnae. Interestingly, there is also a transient premorphogenetic expression of Distal-less in the second antennal and second maxillary segment, although no appendages are eventually formed in these segments. The dachshund gene is apparently involved both in PD patterning as well as in sensory organ development in the antenna, maxilla, and mandible. Strong dachshund expression is specifically correlated with the tooth-like part of the mandible, a feature that is shared with other mandibulate arthropods. homothorax is expressed in the proximal and medial parts of the legs, while extradenticle RNA is only seen in the proximal region. This overlap of expression corresponds to the functional overlap between extradenticle and homothorax in Drosophila. PMID- 12885559 TI - Stage-specific regulation of caspase activity in drosophila oogenesis. AB - In Drosophila oogenesis, the programmed cell death of germline cells occurs predominantly at three distinct stages. These cell deaths are subject to distinct regulatory controls, as cell death during early and midoogenesis is stress induced, whereas the cell death of nurse cells in late oogenesis is developmentally regulated. In this report, we show that the effector caspase Drice is activated during cell death in both mid- and late oogenesis, but that the level and localization of activity differ depending on the stage. Active Drice formed localized aggregates during nurse cell death in late oogenesis; however, active Drice was found more ubiquitously and at a higher level during germline cell death in midoogenesis. Because Drice activity was limited in late oogenesis, we examined whether another effector caspase, Dcp-1, could drive the unique morphological events that occur normally in late oogenesis. We found that premature activation of the effector caspase, Dcp-1, resulted in a disappearance of filamentous actin, rather than the formation of actin bundles, suggesting that Dcp-1 activity must also be restrained in late oogenesis. Overexpression of the caspase inhibitor DIAP1 suppressed cell death induced by Dcp-1 but had no effect on cell death during late oogenesis. This limited caspase activation in dying nurse cells may prevent destruction of the nurse cell cytoskeleton and the connected oocyte. PMID- 12885560 TI - Dissecting Drosophila embryonic brain development using photoactivated gene expression. AB - The Drosophila brain is generated by a complex series of morphogenetic movements. To better understand brain development and to provide a guide for experimental manipulation of brain progenitors, we created a fate map using photoactivated gene expression to mark cells originating within specific mitotic domains and time-lapse microscopy to dynamically monitor their progeny. We show that mitotic domains 1, 5, and 9 give rise to discrete cell populations within specific regions of the brain. Two novel observations were that the antennal sensory system, composed of four disparate cell clusters, arose from mitotic domain 5 and that mitotic domain B produced glial cells, while neurons were produced from mitotic domains 1, 5, and 9. Time-lapse analysis of marked cells showed complex mitotic and migratory patterns for cells derived from these mitotic domains. Photoactivated gene expression was also used either to kill, to induce ectopic divisions, or to alter cell fate. This revealed that deficits were not repopulated, while ectopic cells were removed and extra glia were tolerated. PMID- 12885561 TI - Coordination of BMP-3b and cerberus is required for head formation of Xenopus embryos. AB - Bone morphogenetic proteins (BMPs) and their antagonists are involved in the axial patterning of vertebrate embryos. We report that both BMP-3b and BMP-3 dorsalize Xenopus embryos, but act as dissimilar antagonists within the BMP family. BMP-3b injected into Xenopus embryos triggered secondary head formation in an autonomous manner, whereas BMP-3 induced aberrant tail formation. At the molecular level, BMP-3b antagonized nodal-like proteins and ventralizing BMPs, whereas BMP-3 antagonized only the latter. These differences are due to divergence of their pro-domains. Less BMP-3b than BMP-3 precursor is proteolytically processed in embryos. BMP-3b protein associated with a monomeric form of Xnrl, a nodal-like protein, whereas BMP-3 did not. These molecular features are consistent with their expression profiles during Xenopus development. XBMP-3b is expressed in the prechordal plate, while xBMP-3 is expressed in the notochord. Using antisense morpholino oligonucleotides, we found that the depletion of both xBMP-3b and cerberus, a head inducer, caused headless Xenopus embryos, whereas the depletion of both xBMP-3 and cerberus affected the size of the somite. These results revealed that xBMP-3b and cerberus are essential for head formation regulated by the Spemann organizer, and that xBMP-3b and perhaps xBMP-3 are involved in the axial patterning of Xenopus embryos. PMID- 12885562 TI - A genetic analysis of axon guidance in the C elegans pharynx. AB - We wish to understand how the trajectories of the twenty pharyngeal neurons of C. elegans are established. In this study we focused on the two bilateral M2 pharyngeal motorneurons, which each have their cell body located in the posterior bulb and send one axon through the isthmus and into the metacorpus. We used a GFP reporter to visualize these neurons in cell-autonomous and cell-non-autonomous axon guidance mutant backgrounds, as well as other mutant classes. Our main findings are: 1). Mutants with impaired growth cone functions, such as unc-6, unc 51, unc-73 and sax-3, often exhibit abnormal terminations and inappropriate trajectories at the distal ends of the M2 axons, i.e. within the metacorpus; and 2). Growth cone function mutants never exhibit abnormalities in the proximal part of the M2 neuron trajectories, i.e. between the cell body and the metacorpus. Our results suggest that the proximal and distal trajectories are established using distinct mechanisms, including a growth cone-independent process to establish the proximal trajectory. We isolated five novel mutants in a screen for worms exhibiting abnormal morphology of the M2 neurons. These mutants define a new gene class designated mnm (M neuron morphology abnormal). PMID- 12885563 TI - Gestation stage-dependent intrauterine trophoblast cell invasion in the rat and mouse: novel endocrine phenotype and regulation. AB - Trophoblast cell invasion into the uterine wall is characteristic of hemochorial placentation. In this report, we examine trophoblast cell invasion in the rat and mouse, the endocrine phenotype of invasive trophoblast cells, and aspects of the regulation of trophoblast cell invasion. In the rat, trophoblast cells exhibit extensive interstitial and endovascular invasion. Trophoblast cells penetrate through the decidua and well into the metrial gland, where they form intimate associations with the vasculature. Trophoblast cell invasion in the mouse is primarily interstitial and is restricted to the mesometrial decidua. Both interstitial and endovascular rat trophoblast cells synthesize a unique set of prolactin (PRL)-like hormones/cytokines, PRL-like protein-A (PLP-A), PLP-L, and PLP-M. Invading mouse trophoblast cells also possess endocrine activities, including the expression of PLP-M and PLP-N. The trafficking of natural killer (NK) cells and trophoblast cells within the mesometrial uterus is reciprocal in both the rat and mouse. As NK cells disappear from the mesometrial compartment, a subpopulation of trophoblast cells exit the chorioallantoic placenta and enter the decidua. Furthermore, the onset of interstitial trophoblast cell invasion is accelerated in mice with a genetic deficiency of NK cells, Tg epsilon 26 mice, implicating a possible regulatory role of NK cells in trophoblast cell invasion. Additionally, the NK cell product, interferon-gamma (IFNgamma), inhibits trophoblast cell outgrowth, and trophoblast cell invasion is accelerated in mice with a genetic deficiency in the IFNgamma or the IFNgamma receptor. In summary, trophoblast cells invade the uterine wall during the last week of gestation in the rat and mouse and possess a unique endocrine phenotype, and factors present in the uterine mesometrial compartment modulate their invasive behavior. PMID- 12885564 TI - Arrested apoptosis of nurse cells during Hydra oogenesis and embryogenesis. AB - During Hydra oogenesis, an aggregate of germ cells differentiates into one oocyte and thousands of nurse cells. Nurse cells display a number of features typical of apoptotic cells and are phagocytosed by the growing oocyte. Yet, these cells remain unchanged in morphology and number until hatching of the polyp, which can occur up to 12 months later. Treatments with caspase inhibitors can block oocyte development during an early phase of oogenesis, but not after nurse cell phagocytosis has taken place, indicating that initiation of nurse cell apoptosis is essential for oocyte development. The genomic DNA of the phagocytosed nurse cells in the oocyte and embryo shows large-scale fragmentation into 8- to 15-kb pieces, but there is virtually none of the internucleosomal degradation typically seen in apoptotic cells. The arrested nurse cells exhibit high levels of peroxidase activity and are prevented from entering the lysosomal pathway. After hatching of the polyp, apoptosis is resumed and the nurse cells are degraded within 3 days. During this final stage, nurse cells become TUNEL-positive and enter secondary lysosomes in a strongly degraded state. Our results suggest that nurse cell apoptosis consists of caspase-dependent and caspase-independent phases. The independent phase can be arrested at an advanced stage for several months, only to resume after the primary polyp hatches. PMID- 12885565 TI - Charting the Drosophila neuropile: a strategy for the standardised characterisation of genetically amenable neurites. AB - Insect neurons are individually identifiable and have been used successfully to study principles of the formation and function of neuronal circuits. In the fruitfly Drosophila, studies on identifiable neurons can be combined with efficient genetic approaches. However, to capitalise on this potential for studies of circuit formation in the CNS of Drosophila embryos or larvae, we need to identify pre- and postsynaptic elements of such circuits and describe the neuropilar territories they occupy. Here, we present a strategy for neurite mapping, using a set of evenly distributed landmarks labelled by commercially available anti-Fasciclin2 antibodies which remain comparatively constant between specimens and over developmental time. By applying this procedure to neurites labelled by three Gal4 lines, we show that neuritic territories are established in the embryo and maintained throughout larval life, although the complexity of neuritic arborisations increases during this period. Using additional immunostainings or dye fills, we can assign Gal4-targeted neurites to individual neurons and characterise them further as a reference for future experiments on circuit formation. Using the Fasciclin2-based mapping procedure as a standard (e.g., in a common database) would facilitate studies on the functional architecture of the neuropile and the identification of candiate circuit elements. PMID- 12885566 TI - Ff1b is required for the development of steroidogenic component of the zebrafish interrenal organ. AB - The zebrafish ftz-f1 gene, ff1b, is activated in two cell clusters lateral to the midline in the trunk during late embryogenesis. These cell clusters coalesce to form a discrete organ at around 30 hpf, which then begins to acquire a steroidogenic identity as evidenced by the expression of the steroidogenic enzyme genes, cyp11a and 3beta-hsd. The migration of the cell clusters to the midline is impaired in zebrafish midline signaling mutants. Knockdown of Ff1b activity by antisense ff1b morpholino oligonucleotide (ff1bMO) leads to phenotypes that are consistent with impaired osmoregulation. Injection of ff1bMO was also shown to downregulate the expression of cyp11a and 3beta-hsd. Histological comparison of wild-type and ff1b morphants at various embryonic and juvenile stages revealed the absence of interrenal tissue development in ff1b morphants. The morphological defects of ff1b morphants could be mimicked by treatment with aminoglutethimide, an inhibitor of de novo steroid synthesis. Based on these data, we propose that ff1b is required for the development of the steroidogenic tissue of the interrenal organ. PMID- 12885567 TI - MEI-1/katanin is required for translocation of the meiosis I spindle to the oocyte cortex in C elegans. AB - In most animals, successful segregation of female meiotic chromosomes involves sequential associations of the meiosis I and meiosis II spindles with the cell cortex so that extra chromosomes can be deposited in polar bodies. The resulting reduction in chromosome number is essential to prevent the generation of polyploid embryos after fertilization. Using time-lapse imaging of living Caenorhabditis elegans oocytes containing fluorescently labeled chromosomes or microtubules, we have characterized the movements of meiotic spindles relative to the cell cortex. Spindle assembly initiated several microns from the cortex. After formation of a bipolar structure, the meiosis I spindle translocated to the cortex. When microtubules were partially depleted, translocation of the bivalent chromosomes to the cortex was blocked without affecting cell cycle timing. In oocytes depleted of the microtubule-severing enzyme, MEI-1, spindles moved to the cortex, but association with the cortex was unstable. Unlike translocation of wild-type spindles, movement of MEI-1-depleted spindles was dependent on FZY 1/CDC20, a regulator of the metaphase/anaphase transition. We observed a microtubule and FZY-1/CDC20-dependent circular cytoplasmic streaming in wild-type and mei-1 mutant embryos during meiosis. We propose that, in mei-1 mutant oocytes, this cytoplasmic streaming is sufficient to drive the spindle into the cortex. Cytoplasmic streaming is not the normal spindle translocation mechanism because translocation occurred in the absence of cytoplasmic streaming in embryos depleted of either the orbit/CLASP homolog, CLS-2, or FZY-1. These results indicate a direct role of microtubule severing in translocation of the meiotic spindle to the cortex. PMID- 12885568 TI - Drosophila filamin is required for follicle cell motility during oogenesis. AB - The Filamin family of actin binding proteins is required to maintain cell shape and promote cell locomotion. Using the Drosophila ovary, we provide a detailed description of Filamin-deficient cells during morphogenesis. Reduced expression of Filamin in follicle cells causes defects in the initial encapsulation of germline cysts and in the migration of border cells through the germline cyst. However, follicle cell morphogenesis is unaffected by point mutations that produce truncated Filamin proteins. In addition, mutant follicle cell movements can be partially rescued by a transgene encoding only the actin-binding domain and the first six filamin repeats. These data show that Filamin function in cell motility can be provided by a truncated Filamin protein that resembles Dictyostelium Actin Binding Protein-120. PMID- 12885569 TI - Essential embryonic roles of the CKI-1 cyclin-dependent kinase inhibitor in cell cycle exit and morphogenesis in C elegans. AB - Following a phase of rapid proliferation, cells in developing embryos must decide when to cease division and then whether to survive and differentiate or instead undergo programmed death. In screens for genes that regulate embryonic patterning of the endoderm in Caenorhabditis elegans, we identified overlapping chromosomal deletions that define a gene required for these decisions. These deletions result in embryonic hyperplasia in multiple somatic tissues, excessive numbers of cell corpses, and profound defects in morphogenesis and differentiation. However, cell cycle arrest of the germline is unaffected. Cell lineage analysis of these mutants revealed that cells that normally stop dividing earlier than their close relatives instead undergo an extra round of division. These deletions define a genomic region that includes cki-1 and cki-2, adjacent genes encoding members of the Cip/Kip family of cyclin-dependent kinase inhibitors. cki-1 alone can rescue the cell proliferation, programmed cell death, and differentiation and morphogenesis defects observed in these mutants. In contrast, cki-2 is not capable of significantly rescuing these phenotypes. RNA interference of cki-1 leads to embryonic lethality with phenotypes similar to, or more severe than, the deletion mutants. cki-1 and -2 gene reporters show distinct expression patterns; while both are expressed at around the time that embryonic cells exit the cell cycle, cki-2 also shows marked expression starting early in embryogenesis, when rapid cell division occurs. Our findings demonstrate that cki-1 activity plays an essential role in embryonic cell cycle arrest, differentiation and morphogenesis, and suggest that it may be required to suppress programmed cell death or engulfment of cell corpses. PMID- 12885570 TI - High cholesterol affects platelet APP processing in controls and in AD patients. AB - Alzheimer disease (AD) is characterised by a decrease of platelet Amyloid Precursor Protein forms ratio (APPr), which parallels symptoms' severity. Recent studies have suggested that cholesterol might play a role in the pathophysiology of AD by modulating Abeta production. Aim of this study was to evaluate the relationship between serum cholesterol levels and platelet APP processing in controls and AD. Sixty AD patients and 45 age-matched controls (CTRL) were investigated. Neuropsychological assessment, cholesterol dosage and APP forms' evaluation were performed on each subject. CTRL showed lower serum cholesterol levels compared to AD (P<0.01) and higher mean APPr scores (P<0.0001). Hypercholesterolaemic AD patients showed lower APPr scores compared to normocholesterolaemic AD patients matched for disease severity (0.31+/-0.16 versus 0.45+/-0.28; P<0.05), since the early stage of the disease. In AD, cholesterol levels influence APPr independently of disease severity. These findings confirm the association between cholesterol and AD, and suggest that in vivo cholesterol affects APP processing by interfering with its maturation. PMID- 12885571 TI - Lovastatin enhances Abeta production and senile plaque deposition in female Tg2576 mice. AB - A recent clinical study showed that statins, which are inhibitors of cholesterol biosynthesis pathway, reduced the prevalence of Alzheimer's disease (AD). Animal studies that have employed high cholesterol diet indicate significant relationship between cholesterol level and senile plaque deposition. Here, we investigated the effects of lovastatin on beta-amyloid production and senile plaque deposition in an animal model of AD (Tg2576 mice). As expected, lovastatin treatment reduced plasma cholesterol level in both male and female mice. However, lovastatin enhanced the amounts of beta-amyloid and other beta-secretase derived peptides in females, but not in males. Likewise, lovastatin increased the number of plaques in the hippocampus and cortex of females, but not in males. Lovastatin did not change the amounts of full-length or alpha-secretase processed amyloid precursor protein (APP), or presenilin 1 (PS1) in either sex. Thus, lovastatin lowers cholesterol level in both genders, but enhances beta-amyloid production and senile plaque deposition only in brains of female Tg2576 mice. Our results suggest that low plasma cholesterol levels might be a risk factor for AD in females. PMID- 12885572 TI - Alterations in beta-amyloid production and deposition in brain regions of two transgenic models. AB - Mutations in the amyloid precursor protein (APP) gene are associated with altered production and deposition of amyloid beta (Abeta) peptide in the Alzheimer's disease (AD) brain. The pathways that regulate APP processing, Abeta production and Abeta deposition in different tissues and brain regions remain unclear. To address this, we examined levels of various APP processing products as well as Abeta deposition in a genomic-based (R1.40) and a cDNA-based (Tg2576) transgenic mouse model of AD. In tissues, only brain generated detectable levels of the penultimate precursor to Abeta, APP C-terminal fragment-beta. In brain regions, holoAPP levels remained constant, but ratios of APP C-terminal fragments and levels of Abeta differed significantly. Surprisingly, cortex had the lowest steady-state levels of Abeta compared to other brain regions. Comparison of Abeta deposition in Tg2576 and R1.40 animals revealed that R1.40 exhibited more abundant deposition in cortex while Tg2576 exhibited extensive deposition in the hippocampus. Our results suggest that AD transgenic models are not equal; their unique characteristics must be considered when studying AD pathogenesis and therapies. PMID- 12885573 TI - Two novel presenilin-1 mutations (Y256S and Q222H) are associated with early onset Alzheimer's disease. AB - Mutations in the gene encoding presenilin 1 (PS-1) account for 50% of early-onset familial Alzheimer's disease (EOFAD) cases. In this study, we identified two missense mutations in the coding sequence of the presenilin (PS-1) gene in two EOFAD pedigrees. AD was confirmed in one pedigree by autopsy. Mutation analysis of PCR products amplified from genomic DNA templates showed two novel PS-1 mutations resulting in Gln222His and Tyr256Ser. The two novel mutations are located within predicted transmembrane domains five (TM-5) and six (TM-6), respectively, and are associated with very early ages of onset. The Tyr256Ser is associated with one of the youngest age of AD onset, 25 years, which is consistent with a drastic change in function of the altered PS-1 protein. A morphometric analysis of the cortical degenerative changes of the Tyr256Ser case, showed severe involvement of the primary motor cortex, which correlated well with the pyramidal changes, including tetraspasticity. Immunoblot analysis showed the Tyr256Ser case had the greatest expression of Abeta(1-40) and Abeta(1-42), which was confirmed by ELISA, compared to other PS-1 mutant FAD cases and age-matched controls and, thus, contributes to the severity of the disease pathology. PMID- 12885574 TI - 3-Nitrotyrosine immunoreactivity in olfactory receptor neurons of patients with Alzheimer's disease: implications for impaired odor sensitivity. AB - Olfactory sensory function is impaired in patients with the diagnosis of probable Alzheimer's disease (AD) compared to elderly controls, and the olfactory epithelium (OE) of AD patients exhibits several pathological changes characteristic of the AD brain. To confirm that the populations from whom our postmortem tissues are obtained exhibit similar decrements in sensory function, threshold testing was performed; probable AD patients had significantly higher olfactory thresholds than controls. To determine if oxidative stress contributes to decreased olfactory function in AD, we localized 3-nitrotyrosine (3-NT) immunoreactivity in OE obtained postmortem from patients with neuropathologically confirmed AD and age-matched controls with brains free of significant neurodegenerative pathology. In AD patients, immunoreactivity was localized in olfactory receptor neurons (ORNs), including dendritic knobs where ion channels that participate in sensory transduction are located, suggesting a direct mechanism for olfactory impairment. In controls, immunoreactivity occurred in blood vessel endothelium, suggesting age-related vascular dysfunction. Immunohistochemistry for CD68, a macrophage scavenger receptor, demonstrated activated macrophages, a source of free radicals contributing to 3-NT formation, in the OE of AD patients but not controls. These results demonstrate increased oxidative stress and modification of ORN proteins that may contribute directly to olfactory impairment in AD patients. PMID- 12885575 TI - Response-monitoring dysfunction in aging and Alzheimer's disease: an event related potential study. AB - Executive control is a broad-reaching function that includes response monitoring and is likely implemented in the frontal lobes. Age- and dementia-related changes in response-monitoring were assessed during a Picture-Name Verification Task, using response-synchronized event-related potential (ERP) markers of response monitoring: the centrally oriented error-related negativity (ERN); the smaller and more frontally-oriented correct-response negativity (CRN); and the positivity associated with errors (Pe), a marker of error awareness. These were recorded from 10 younger and 10 older healthy controls, as well as 12 Alzheimer's disease (AD) patients. Although the older and younger controls showed equivalent accuracy, error awareness (Pe), and relative ERN>CRN amplitude, aging was associated with slower behavioral responses and decreased ERN amplitude. Although dementia was associated with decreased accuracy, decreased ERN, and a loss of relative ERN>CRN amplitude, error awareness (Pe) remained somewhat intact in AD patients. In AD patients, CRN amplitude was affected by item certainty (assessed a week earlier), being larger to items that were idiosyncratically difficult for patients to name. PMID- 12885576 TI - Cell cycle aberrations by alpha-synuclein over-expression and cyclin B immunoreactivity in Lewy bodies. AB - alpha-Synuclein is a presynaptic protein that accumulates abnormally in Lewy bodies of Parkinson's disease (PD) and dementia with Lewy bodies (DLB). Its physiological function and role in neuronal death remain poorly understood. Recent immunohistochemical studies suggest that cell cycle-related phenomena may play a role in the pathogenesis of Alzheimer's disease and perhaps other neurodegenerative disorders. In this investigation, we examined the effects of alpha-synuclein expression levels on cell cycle indices in PC12 cells engineered to conditionally induce alpha-synuclein expression upon withdrawal of doxycycline. Over-expression of alpha-synuclein resulted in enhanced proliferation rate and enrichment of cells in the S phase of the cell cycle. This was associated with increased accumulation of the mitotic factor cyclin B and down-regulation of the tumor suppressor retinoblastoma 2. Additionally, ERK1/2, key molecules in proliferation signaling, were highly phosphorylated. Immunohistochemical studies on postmortem brains revealed intense cyclin B immunoreactivity in Lewy bodies in cases with DLB and to a lesser extent in PD. We propose that elevated expression of alpha-synuclein causes changes in cell cycle regulators through ERK activation leading to apoptosis of postmitotic neurons. These changes in cell cycle proteins are also associated with ectopic expression of cyclin B in Lewy bodies. PMID- 12885577 TI - Dopamine neurotoxicity: age-dependent behavioral and histological effects. AB - The oxidative stress (OS) theory has implicated the involvement of reactive oxygen species (ROS) in both aging and age-dependent neurodegenerative diseases. The dopaminergic system is particularly vulnerable to ROS, and dopamine (DA) itself can be an endogenous source of ROS. The present study evaluated the hypothesis that DA-induced toxicity is age-dependent, and tested the behavioral and histological correlates of DA neurotoxicity in aging. Young (6 months) and middle-aged (15 months) rats were chronically treated with DA in the substantia nigra (SN, 1 micromol/2 microl vehicle per side/day/5 days) and were subsequently examined for changes in motor function and histology. The neurotoxic effect of DA treatment was an age-dependent effect, as middle-aged animals that received DA infusions in the SN were more impaired than their age-matched controls, especially on tasks that involved greater sensory-motor coordination, whereas young animals that received DA behaved similarly to their age-matched controls. The behavioral effects noted were accompanied by a loss of the tyrosine hydroxylase phenotype in substantia nigra. However, selective neurodegeneration was not noted in the SN of the treated animals, nor was a selective iron deposition noted at the site of injection. These results suggest that a neurochemical deficit and not cell loss per se within the nigrostriatal system underlies the motor behavioral deficits observed in the middle-aged rats. PMID- 12885578 TI - Normal pressure hydrocephalus triggers intrathecal production of TNF-alpha. AB - Normal pressure hydrocephalus (NPH) is associated with periventricular white matter lesions and demyelination. The aim of the present study was to examine the cerebrospinal fluid (CSF) levels of tumor necrosis factor-alpha (TNF-alpha), a proinflammatory cytokine mediating myelin damage, in patients with NPH. TNF-alpha levels were analyzed by ELISA and measured before and after shunt operation in 35 patients with NPH. The levels of this cytokine were related to the symptomatology and to magnetic resonance imaging (MRI) verified white matter lesions. They were also related to intrathecal levels of sulfatide, a marker for white matter degradation and to levels of neurofilament, a marker for neuronal degeneration. The preoperative levels of TNF-alpha were increased in the CSF of NPH patients compared to controls, and correlated to the levels of sulfatide. The intrathecal TNF-alpha levels were higher in NPH patients with impairment of wakefulness than in those without this symptom. The preoperative TNF-alpha levels were significantly correlated to the improvement of psychometrical test scores, and of wakefulness and to the overall improvement of the patients following shunt operation. Importantly, shunt operation led to complete disappearance of intrathecal TNF-alpha. We conclude that NPH is correlated with intrathecal TNF alpha production being reversed following shunt operation in parallel with the clinical improvement. The positive correlation between preoperative TNF-alpha and sulfatide levels in the CSF suggest that intrathecal TNF-alpha may contribute to the damage of the white matter known to occur in patients with NPH. PMID- 12885579 TI - Age-related differences in MK-801- and amphetamine-induced locomotor and stereotypic activities of rats. AB - Changes in locomotor and stereotypic activities induced by an i.p. injection of either (+)-5-methyl-10,11-dihydro-5H-dibenzo(a,d)-cycloheptan-5,10-imine maleate (dizocilpine or MK-801; 0.3 mg/kg) or D-amphetamine sulfate (AMPH; 1.5 mg/kg) were studied in male Mill Hill hooded rats of different age. The following age groups of animals were considered: 28-30 postnatal day (PND)-old rats (peripubertal), 48-50 PND-old (pubertal), 3-month-old (adults), 12-month-old (middle-aged) and 24-month-old (aged). The motor response was measured by an automated animal activity measuring system. The obtained results showed that: (1) in contrast to AMPH, MK-801 induced more pronounced increases of both locomotor and stereotypic activities in peripubertal and pubertal than in adult and aged rats; (2) AMPH induced the same locomotor and stereotypic activity increase in pubertal, adult and middle-aged rats; (3) both AMPH and MK-801 led to a senescence-related decrease of motor activity. These data suggest that the balance of the glutamatergic and dopaminergic systems changes during aging. Such a change is important in understanding schizophrenia and the motor system decline observed in the later stages of life. PMID- 12885580 TI - Preliminary evidence that estrogen protects against age-related hippocampal atrophy. AB - Few studies have examined gender differences in hippocampal volumes, and the potential effect of estrogen on these measures has not been well studied. We used MRI to measure hippocampal volumes in elderly Mexican American men and women subjects in order to determine if there were gender differences and if estrogen replacement therapy (ERT) had an effect on hippocampal volume in postmenopausal women. MRI measures of hippocampal volumes (normalized to intracranial volume) were compared in 59 women and 38 men. Further comparisons were made between men subjects, women subjects taking ERT, and women subjects not taking ERT. There were no significant effects of gender on normalized hippocampal volumes. However, women subjects taking ERT had larger right hippocampal volumes than women subjects not taking ERT and larger anterior hippocampal volumes than men subjects and women subjects not taking ERT. These findings suggest a neuroprotective effect of estrogen. PMID- 12885581 TI - Differential effects of estrogen in the injured forebrain of young adult and reproductive senescent animals. AB - Estrogen attenuates neural damage resulting from a variety of experimental injury models in adult female rats. To determine whether estrogens neuroprotective actions are age-specific, the present study compared the effects of estrogen on young adult and reproductive senescent animals subject to excitotoxic injury to the forebrain. NMDA was injected bilaterally into the olfactory bulbs of estrogen and placebo-replaced young adult and reproductive senescent animals. Lysates of the olfactory bulb and its basal forebrain afferent, the horizontal limb of the diagonal band of Broca (hlDBB), harvested 24h later were analyzed for expression of IL-1beta, IL-10, and nerve growth factor (NGF). NMDA injections resulted in local activation of microglia and an increase in IL-1beta. Estrogen replacement decreased IL-1beta expression in young adult females, but paradoxically enhanced its expression in reproductive senescent females. Furthermore, bulb injury increased IL-1beta production in the hlDBB of reproductive senescent animals although estrogen replacement was able to suppress lesion-induced expression of this cytokine. In both, the olfactory bulb and hlDBB, constitutive expression of the anti-inflammatory cytokine IL-10 was significantly higher while that of NGF was almost 50% lower in senescent animals as compared to young adults, indicating that aging preferentially altered Th2-type secretions. The present findings are consistent with our earlier observations that estrogen does not exert trophic effects in the aging forebrain and supports the hypothesis that estrogen treatment to reproductive senescent females may exacerbate neural injury. PMID- 12885582 TI - An increase of sigma receptors in the aged monkey brain. AB - We evaluated in vivo the effect of aging on the sigma(1) receptors in the monkey brain by the quantitative analysis of the binding of [11C]SA4503 to sigma(1) receptors with positron emission tomography. Based on a three-compartment model, the influx rate constant K(1) of [11C]SA4503 from plasma to brain across the blood-brain barrier in all 10 regions investigated became smaller in the aged monkeys (20-28 years old, n=5) than in the young adult monkeys (4-8 years old, n=5), but the reduction was not significant due to the individual differences. On the other hand, the binding potential, which was calculated as the ratio of the association rate constant k(3) to the dissociation rate constant k(4) for the binding of [11C]SA4503 to sigma(1) receptors in the brain, significantly increased in nine of the brain regions of the aged monkeys to the 160-210% levels of the young monkeys. We concluded that the sigma(1) receptor binding sites increased in the aging process of the monkey brain. PMID- 12885583 TI - Age-related changes in cerebral lactate metabolism in sleep-disordered breathing. AB - Thirty-one patients, aged 22-71 years, with nocturnal apneic episodes and/or habitual snoring were studied with magnetic resonance spectroscopy (MRS) and diagnostic polysomnography separately to determine whether accumulation of lactate caused by cerebral hypoxia during sleep is associated with sleep disordered breathing (SDB), aging and co-morbidities related to SDB. Eight proton magnetic resonance spectra for sleep and two for periods of arousal were obtained from the right centrum semiovale. All patients were evaluated for the presence or absence of co-morbidities including hypertension, cardiac disease, diabetes mellitus, and hyperlipidemia. Significant lactate signals were found in seven patients with obstructive sleep apnea-hypopnea syndrome (OSAHS) during sleep periods, and none during periods of arousal. Aging was significantly related to the presence or absence of significant lactate signals during sleep periods as determined by logistic regression analysis (beta=0.2480; 95% confidence interval, 0.0905-0.5094; P=0.0001). Apnea index (AI), apnea-hypopnea index (AHI), and minimum value of peripheral oxyhemoglobin saturation each significantly interacted with age (P=0.0081, 0.0284, and 0.0302, respectively). Our findings suggest that SDB combined with aging is related to accumulation of lactate during sleep. PMID- 12885584 TI - Oxidative and nitrosative events in asthma. AB - Asthma affects over 15 million individuals in the United States, with over 1.5 million emergency room visits, 500,000 hospitalizations, and 5500 deaths each year, many of which are children. Airway inflammation is the proximate cause of the recurrent episodes of airflow limitation in asthma. Research applying molecular biology, chemistry, and cell biology to human asthma and model systems of asthma over the last decade has revealed that numerous biologically active proinflammatory mediators lead to increased production of reactive oxygen species (ROS) and the gaseous molecule nitric oxide (NO). Persistently increased ROS and NO in asthma lead to reactive nitrogen species (RNS) formation and subsequent oxidation and nitration of proteins, which may cause alterations in protein function that are biologically relevant to airway injury/inflammation. Eosinophil peroxidase and myeloperoxidase, leukocyte-derived enzymes, amplify oxidative events and are another enzymatic source of NO-derived oxidants and nitrotyrosine formation in asthma. Concomitant with increased generation of oxidative and nitrosative molecules in asthma, loss of protective antioxidant defense, specifically superoxide dismutase (SOD), contributes to the overall toxic environment of the asthmatic airway. This review discusses the rapidly accruing data linking oxidative and nitrosative events as critical participants in the acute and chronic inflammation of asthmatic airways. PMID- 12885585 TI - ROS in the local and systemic pathogenesis of COPD. AB - An imbalance between oxidants and antioxidants is proposed in the pathogenesis of COPD. Potential alterations responsible for an imbalance in oxidant production and intra- and extracellular antioxidant defense systems are discussed with respect to COPD-related changes in the pulmonary compartment. In line with the current view of COPD as a disease with multiple systemic consequences, there is increasing evidence that imbalances in the redox milieu extend beyond the diseased lung in COPD patients. Skeletal muscle dysfunction is often observed in COPD and may result from imbalances in the redox environment of skeletal muscle. Potential triggers of oxidative stress in the muscle compartment include inflammation and hypoxia, and local sources of reactive oxygen and nitrogen species are discussed, as well the mechanisms by which skeletal muscle trophical state, contractility and fatigability may be affected by oxidative stress, resulting in skeletal muscle dysfunction. PMID- 12885586 TI - Extracellular superoxide dismutase in biology and medicine. AB - Accumulated evidence has shown that reactive oxygen species (ROS) are important mediators of cell signaling events such as inflammatory reactions (superoxide) and the maintenance of vascular tone (nitric oxide). However, overproduction of ROS such as superoxide has been associated with the pathogenesis of a variety of diseases including cardiovascular diseases, neurological disorders, and pulmonary diseases. Antioxidant enzymes are, in part, responsible for maintaining low levels of these oxygen metabolites in tissues and may play key roles in controlling or preventing these conditions. One key antioxidant enzyme implicated in the regulation of ROS-mediated tissue damage is extracellular superoxide dismutase (EC-SOD). EC-SOD is found in the extracellular matrix of tissues and is ideally situated to prevent cell and tissue damage initiated by extracellularly produced ROS. In addition, EC-SOD is likely to play an important role in mediating nitric oxide-induced signaling events, since the reaction of superoxide and nitric oxide can interfere with nitric oxide signaling. This review will discuss the regulation of EC-SOD and its role in a variety of oxidant-mediated diseases. PMID- 12885587 TI - Thyroid hormone-induced oxidative stress triggers nuclear factor-kappaB activation and cytokine gene expression in rat liver. AB - Nuclear factor-kappaB (NF-kappaB) is a redox-sensitive factor responsible for the transcriptional activation of cytokine-encoding genes. In this study, we show that 3,3,5-triiodothyronine (T(3)) administration to rats activates hepatic NF kappaB, as assessed by electrophoretic mobility shift assay. This response coincides with the onset of calorigenesis and enhancement in hepatic respiration, and is suppressed by the antioxidants alpha-tocopherol and N-acetylcysteine or by the Kupffer cell inactivator gadolinium chloride. Livers from hyperthyroid rats with enhanced NF-kappaB DNA-binding activity show induced mRNA expression of the NF-kappaB-responsive genes for tumor necrosis factor-alpha (TNF-alpha) and interleukin- (IL-) 10, as evidenced by reverse transcription-polymerase chain reaction assay, which is correlated with increases in the serum levels of the cytokines. T(3) also increased the hepatic levels of mRNA for IL-1alpha and those of IL-1alpha in serum, with a time profile closely related to that of TNF-alpha. It is concluded that T(3)-induced oxidative stress enhances the DNA-binding activity of NF-kappaB and the NF-kappaB-dependent expression of TNF-alpha and IL 10 genes. PMID- 12885589 TI - Pretreatment growth environments alter the sensitivity of tumor cells to cytotoxic agents. AB - The effects of pretreatment growth conditions on the sensitivity of tumor cells to various cytotoxic agents were investigated using murine Ehrlich ascites tumor cells grown in two different environments. The tumor cells adapted to grow in the peritoneal cavity of mice were found to be more sensitive to ionizing radiation, oxygen toxicity, doxorubicin, and bleomycin than tumor cells adapted to grow in vitro. However, there was no difference in their sensitivity to 5-fluorouracil. One obvious difference between these two growth environments is oxygen tension; it is between 2.6 and 5.2% (20-40 mmHg) for the peritoneal cavity and 21% (159 mmHg) for the regular tissue culture. To investigate the role of oxygen tension, tumor cells from the peritoneal cavity were grown in tissue culture having either 21% O2 or 4% O2 in the gas phase. Within 4 d, tumor cells that were exposed to 21% O2, but not to 4% O2, in vitro gradually became as resistant to cytotoxic agents as the tumor cells continuously cultured in vitro under 21% O2. It appears that the adaptation of tumor cells to different environments having different partial pressure of oxygen alters their sensitivity not only to oxygen toxicity but also to other cytotoxic agents that damage or kill cells by generating free radicals. PMID- 12885588 TI - Attenuation of leukocyte-endothelium interaction by antioxidant enzymes. AB - This report assessed the effect of overexpressing Cu,Zn superoxide dismutase (SOD) and/or catalase on the interaction of mononuclear cells (MNCs) and endothelial cells (ECs). ECs were obtained from the aorta of wild-type mice and transgenic mice overexpressing Cu,ZnSOD and/or catalase. MNCs were obtained from wild-type mice. Treatment of wild-type ECs with CuSO4-oxidized low-density lipoprotein (oxLDL) significantly elevated the expression of vascular cell adhesion molecule-1 (VCAM-1) and intercellular adhesion molecule-1 (ICAM-1) and increased the adherence of MNCs. Overexpression of Cu,ZnSOD and/or catalase in ECs attenuated the adherence of MNCs and the expression of cell adhesion molecules induced by oxLDL. For example, ECs overexpressing Cu,ZnSOD and/or catalase showed significantly less expression of VCAM-1 and ICAM-1 and less number of adherent MNCs than wild-type ECs. Moreover, ECs overexpressing Cu,ZnSOD and catalase in combination showed significantly less expression of VCAM-1 and ICAM-1 and less number of adherent MNCs than those overexpressing either Cu,ZnSOD or catalase alone. These results suggest that combinational overexpression of Cu,ZnSOD and catalase can reduce the expression of cell adhesion molecules and inhibit the adherence of leukocyte to ECs more efficiently than overexpression of Cu,ZnSOD or catalase alone. PMID- 12885590 TI - Exercise-induced endotoxemia: the effect of ascorbic acid supplementation. AB - Strenuous, long-duration aerobic exercise results in endotoxemia due to increased plasma levels of lipopolysaccharide (LPS) leading to cytokine release, oxidative stress, and altered gastrointestinal function. However, the effect of short-term strenuous aerobic exercise either with or without antioxidant supplementation on exercise-induced endotoxemia is unknown. A significant increase in the concentration of bacterial LPS (endotoxin) was noted in the venous circulation of healthy volunteers following maximal acute aerobic exercise (0.14(-1) pre exercise vs. 0.24(-1) postexercise, p <0.01). Plasma nitrite concentration also increased with exercise (0.09 +/- 0.05 nM x ml(-1) vs. 0.14 +/- 0.01 nM x ml(-1), p <0.05) as did ascorbate free radical levels (0.02 +/- 0.001 vs. 0.03 +/- 0.002 arbitrary units, p <0.05). Oral ascorbic acid supplementation (1000 mg) significantly increased plasma ascorbic acid concentration (29.45 mM x l(-1) to 121.22 mM x l(-1), p <0.05), and was associated with a decrease in plasma LPS and nitrite concentration before and after exercise (LPS: 0.01(-1); nitrite: 0.02 +/- 0.02 nM x ml(-1) vs. 0.02 +/- 0.03 nM x ml(-1)). Ascorbic acid supplementation led to a significant increase in ascorbate free radical levels both before (0.04 +/- 0.01 arbitrary units) and after exercise (0.06 +/- 0.02 arbitrary units, p <0.05). In conclusion, strenuous short-term aerobic exercise results in significant increases in plasma LPS levels (endotoxemia) together with increases in markers of oxidative stress. Supplementation with ascorbic acid, however, abolished the increase in LPS and nitrite but led to a significant increase in the ascorbate radical in plasma. The amelioration of exercise-induced endotoxemia by antioxidant pretreatment implies that it is a free radical-mediated process while the use of the ascorbate radical as a marker of oxidative stress in supplemented systems is limited. PMID- 12885591 TI - Common mechanisms for declines in oxidative stress tolerance and proliferation with aging. AB - Aging is often characterized by reduced stress tolerance and decreased proliferative capacity, but little is known about the effects of aging on signaling pathways important in regulating these responses. Recent studies from our laboratory have implicated impairments in epidermal growth factor receptor (EGFR) signaling and extracellular signal-regulated kinase (ERK) activation to both effects in rat hepatocytes. Here we investigated the responsiveness of hepatocytes derived from young (4-5 months) and aged (24-29 months) mice to proliferative signals (low concentrations of H2O2 and epidermal growth factor [EGF] stimulation), and oxidant injury (high H2O2 concentrations). Old hepatocytes displayed lower levels of DNA synthesis in response to low H(2)O(2) concentrations (5-10 microM) and EGF stimulation, and reduced survival following treatment with high H2O2 concentrations (20-50 microM). Both effects were associated with reduced activation of ERK and diminished phosphorylation of EGFR tyrosine residue 1173. p38 was also activated by H2O2, but to a greater extent in old cells. Pharmacologic inhibition of ERK increased the sensitivity of young cells to H2O2-induced cell death, while inhibition of p38 decreased the sensitivity of old cells. Our findings suggest that impairments in common signaling events underlie age-related declines in proliferative capacity and oxidative stress tolerance in mouse hepatocytes, and that an imbalance in ERK and p38 activities contributes to the greater sensitivity of aged cells to H2O2. PMID- 12885592 TI - Coenzyme Q(10) supplementation inhibits aortic lipid oxidation but fails to attenuate intimal thickening in balloon-injured New Zealand white rabbits. AB - Oxidized lipoproteins are implicated in atherosclerosis, and some antioxidants attenuate the disease in animals. Coenzyme Q(10) (CoQ(10)) in its reduced form, ubiquinol-10, effectively inhibits lipoprotein oxidation in vitro and in vivo; CoQ(10) supplements also inhibit atherosclerosis in apolipoprotein E gene knockout (apoE-/-) mice. Here we tested the effect of dietary CoQ(10) supplements on intimal proliferation and lipoprotein lipid oxidation in balloon-injured, hypercholesterolemic rabbits. Compared to nonsupplemented chow, CoQ(10) supplementation (0.5% and 1.0%, w/w) significantly increased the plasma concentration of CoQ(10) and the resistance of plasma lipids to ex vivo oxidation. CoQ(10) supplements also increased the content of CoQ(10) in the aorta and liver, but not in the brain, skeletal muscle, kidney, and heart. Surprisingly, CoQ(10) supplementation at 1% increased the aortic concentrations of all lipids, particularly triacylglycerols, although it significantly inhibited the proportion of triacylglycerols present as hydroperoxides by > 80%. The observed increase in vessel wall lipid content was reflected in elevated plasma concentrations of cholesterol, cholesteryl esters and triacylglycerols, and hepatic levels of mRNA for 3-hydroxy-3-methylglutaryl-coenzyme A reductase. CoQ(10) supplements did not attenuate lesion formation, assessed by the intima-to media ratio of injured aortic vessels. Thus, like in apoE-/- mice, a high dose of supplemented CoQ(10) inhibits lipid oxidation in the artery wall of balloon injured, hypercholesterolemic rabbits. However, unlike its antiatherosclerosis activity in the mice, CoQ(10) does not inhibit intimal hyperplasia in rabbits, thereby dissociating this disease process from lipid oxidation in the vessel wall. PMID- 12885593 TI - Pro-oxidative activity of nitroxides in their reactions with glutathione. AB - Nitroxides are unreactive towards glutathione in vitro. Interaction of nitroxides with peroxynitrite does not lead to a significant loss of their electron paramagnetic resonance (EPR) signal. However, addition of peroxynitrite to a solution containing glutathione and nitroxides induces an irreversible disappearance of EPR signal of nitroxides and augmentation of glutathione oxidation which is a pro-oxidant effect of these compounds. Nitroxide loss leading to the formation of amine derivatives is initiated by products of glutathione oxidation by peroxynitrite. The pro-oxidant action of nitroxides at micromolar concentrations may be important in view of the proposed use of these compounds as antioxidants. PMID- 12885594 TI - Antioxidant S-allylcysteine prevents gentamicin-induced oxidative stress and renal damage. AB - Acute renal failure (ARF) is a major complication of gentamicin (GM) treatment, which is effective against gram-negative infections. Since experimental evidence suggests a role of reactive oxygen species (ROS) in GM-induced ARF, in this work we studied the effect of a garlic-derived compound, S-allylcysteine (SAC), which is a free radical scavenger, on GM-induced nephrotoxicity. In rats treated with GM (70 mg/kg/12 h/4 days/s.c.), ARF was evident by the: (i) decrease in creatinine clearance and increase in blood urea nitrogen, (ii) decrease in blood glutathione peroxidase (GPx) activity and increase in urinary excretion of N acetyl-beta-D-glucosaminidase and total protein, and (iii) necrosis of proximal tubular cells. These alterations were prevented by SAC treatment (250 mg/kg/i.p. 24 h before the first dose of GM and 125 mg/kg/12 h/4 days along GM-treatment). Furthermore, SAC prevented the GM-induced oxidative stress (protein carbonyl groups) and the decrease in manganese superoxide dismutase (Mn-SOD), GPx, and glutathione reductase (GR) activities in renal cortex. In conclusion, SAC ameliorates the GM-induced ARF by a mechanism related, at least in part, to its ability to decrease oxidative stress and to preserve antioxidant enzymes activity in renal cortex. PMID- 12885595 TI - A template for change and response to work hour restrictions. AB - Surgical program directors are seeking how to best meet the work hour restrictions recently mandated by the American College of Graduate Medical Education. Implementation of an 80-hour work week forces major change to graduate medical education, especially surgical education. Creative restructuring of surgical training is necessary to ensure compliance. Developing an innovative solution to meet these requirements must consider programmatic needs, requiring commitment to a change process. The Department of Surgery at Eastern Virginia Medical School experienced a 5-month strategic planning process that generated the Mendoza plan. This plan uses an every third night call model and a night float model to meet site-specific needs. The specifics of the Mendoza plan protect the cornerstone of surgical education, which is continuity of patient care and resident education. The Mendoza plan, and the process leading to its development, may provide insightful information for other surgical residency programs planning to meet work hour guidelines. PMID- 12885596 TI - Trauma care and the "lethality perspective". PMID- 12885597 TI - Predicting residual lymph node basin disease in melanoma patients with sentinel lymph node metastases. AB - BACKGROUND: The incidence of residual occult disease in nonsentinel lymph nodes (NSLN) after a positive sentinel lymph node (SLN) biopsy in patients with melanoma is relatively low. The purpose of this study is to identify factors that may be predictive of occult NSLN metastases after positive SLN biopsy. METHODS: Fifty-six consecutive melanoma patients with positive sentinel nodes who subsequently underwent complete lymph node dissection (CLND) were evaluated. RESULTS: Only the number of positive SLN predicted the status on NSLN by univariate (P = 0.008) and multivariate (P = 0.028) analyses. None of the other variables (characteristics of SLN metastases, number of draining nodal basins, age, sex, thickness, Clark level, ulceration, number of mitoses/mm(2), histological subtype, and location of the primary) significantly predicted CLND results. CONCLUSIONS: Identifying patients with residual lymph node basin disease remains difficult. Thus, lymph node dissection should be performed in all patients after positive sentinel node biopsy. PMID- 12885598 TI - Sentinel node biopsy prior to neoadjuvant chemotherapy. AB - BACKGROUND: Several studies have explored sentinel lymph node biopsy (SLNB) after neoadjuvant chemotherapy, but false negative rates and the loss of pretreatment nodal staging are limitations. Sentinel lymph node biopsy prior to induction chemotherapy may address both. METHODS: Sentinel lymph node biopsy was performed in clinically node negative patients prior to initiating chemotherapy. Standard level I/II axillary lymph node dissection (ALND) was performed at the time of surgery in those patients who had metastases in the sentinel lymph node (SLN). RESULTS: Twenty-five patients had 26 SLNB prior to the initiation of chemotherapy. The SLN was identified in all cases (100%). Twelve patients (48%) were found to be node negative and did not require axillary node dissection after chemotherapy. Of the patients who were SLN positive and underwent completion ALND, residual nodal disease was identified in 60%. There were no surgical complications or delay of chemotherapy. CONCLUSIONS: Sentinel lymph node biopsy prior to neoadjuvant chemotherapy can avoid the morbidity of ALND without compromising the accuracy of axillary staging. It allows for identification of node positive patients subsequently rendered disease free in the regional nodes, which can assist in planning additional chemotherapy or radiation. PMID- 12885599 TI - New alternatives in the management of gastroesophageal reflux disease. AB - BACKGROUND: To date the mainstay of surgical treatment for gastroesophageal reflux disease (GERD) has been achieved with either open or laparoscopic fundoplication. Several new treatment modalities are attempting to augment the gastroesophageal (GE) junction function by various endoscopic means. METHODS: The Medline database from 1980 to 2002 was searched for studies on endoscopic techniques for antireflux procedures. Product investigators were contacted for data presented in abstract form only. RESULTS: Recent improvements in equipment and technique with excellent long-term follow-up have made laparoscopic Nissen fundoplication the gold standard in antireflux surgery. New techniques include using radiofrequency energy, injection of silicon type polymer and using endoluminal sutures to narrow the gastroesophageal junction. Early results have encouraging aspects, but should be evaluated thoroughly and with caution before widespread use. CONCLUSIONS: Endoscopic treatment of gastroesophageal reflux has future promise. However, more experience and perhaps further improvement in techniques and technology must occur before wide application can be encouraged. PMID- 12885600 TI - Ductal carcinoma in situ with microinvasion. AB - BACKGROUND: Ductal carcinoma in situ (DCIS) accounts for nearly 20% of new breast cancer diagnoses and ductal carcinoma in situ with microinvasion (DCIS-MI) is found in 5% to 10% of DCIS. Controversy exists regarding the appropriate local treatment as well as whether or not examination of the axilla should be performed either by sentinel lymph node biopsy (SLNB), axillary lymph node dissection (ALND) or not at all. METHODS: A MEDLINE search was performed using the keywords ductal carcinoma in situ and microinvasion. Recent articles pertaining to the definition and characterization of DCIS-MI as well as treatment and prognosis were analyzed. CONCLUSIONS: The data at this time demonstrate no survival benefit for patients undergoing mastectomy versus lumpectomy and radiation. Numerous studies demonstrate axillary lymph node involvement to be as high as 20% with DCIS-MI; therefore, we believe that axillary sampling is essential. We recommend SLNB, which is accurate, provides information necessary for staging and treatment, and is associated with less morbidity than traditional ALND. PMID- 12885601 TI - The next stage in the health care economy: aligning the interests of patients, providers, and third-party payers through consumer-driven health care plans. AB - This article reviews employers' attempts over the past 25 years to address the cost and accessibility of health care services for their employees and the effect these efforts have had on U.S. health care delivery. The difficulties in aligning the interests of all parties in a third-party health beneficiary contract are examined. Many employers are considering consumer-driven health care plans as an alternative to managed care plans to both control health care costs and improve employee satisfaction. Such plans differ from fee-for-service and managed care models in terms of the economic alignment of the parties. Consumer-driven plans align the employer's economic interest with the employee/patient, and reduce health benefit costs by providing information, tools, and direct economic incentives to employees for self-management of health care dollars. Because these incentives are designed to reduce the consumption of services, providers are the party left out of economic alignment under the consumer-driven model. PMID- 12885602 TI - Medical student operative experience correlates with a match to a categorical surgical program. AB - BACKGROUND: Interest in general surgical residencies has decreased significantly. Because medical student clerkship experiences may affect specialty preferences, we attempted to determine if the degree of exposure to surgical procedures influenced career choices. METHODS: Operations observed by students who completed the third-year surgical clerkship between 1998 and 1999 were reviewed. These 146 medical students, who matched to residency training programs in March 2000, were then divided into three groups based upon residency fields. Surgical case exposures were then compared between the groups. RESULTS: The total number of operations observed was similar between the groups. However, students who matched into categorical general surgical programs participated in significantly more abdominal and general surgical procedures than those matching in surgical subspecialty or nonsurgical residencies (P < 0.01). CONCLUSIONS: There appears to be a correlation between surgical case exposure during the third-year clerkships and future residency fields. Thus, the degree of exposure to surgical procedures may influence medical student career choices. PMID- 12885603 TI - Diverticulum of the vermiform appendix is associated with chronic abdominal pain. AB - BACKGROUND: Diverticulum of the vermiform appendix is rarely encountered. Previous reports in the literature indicate that appendiceal diverticulum without inflammations are asymptomatic. METHODS: During the past 10 years, 217 patients have been evaluated in our office for chronic lower abdominal pain. Six patients in this group were unusual in that no diagnosis could be made after a complete history, physical examination, and multiple imaging and laboratory studies. Each patient had a normal computed tomography scan of the abdomen, upper endoscopy, colonoscopy, barium enema, and small bowel enteroclysis. Each of the 6 patients underwent an exploratory laparoscopy and appendectomy. RESULTS: All 6 patients in this series had acquired diverticulum of the appendix with inspissated intraluminal secretions without histological evidence of inflammation. The age range was 37 to 57 years. Most patients were male, and most had a single diverticulum projecting into the mesoappendix. All 6 patients had complete resolution of their chronic abdominal pain after the laparoscopic appendectomy. CONCLUSIONS: An unusual cause of mild chronic abdominal pain can be the presence of appendiceal diverticulum with inspissated intraluminal secretions. The evaluation of these patients is usually normal. Exploratory laparoscopy with appendectomy should be the final evaluation for patients with mild chronic abdominal pain. PMID- 12885604 TI - Meckel's diverticulum in an adult gastrointestinal bleed. PMID- 12885605 TI - Evaluation of mammographic and clinical follow-up after 755 stereotactic vacuum assisted breast biopsies. AB - PURPOSE: Stereotactic vacuum-assisted breast biopsy (VB) is a new method that promises high accuracy and reliability. In order to avoid surgery in cases with benign histology the examination must be quality assured and the accuracy should be well established. We present follow-up data of 755 VBs with benign results. METHODS: In all, 984 of 1268 consecutive VBs proved histopathologically benign (lobular carcinoma in situ and atypical ductal hyperplasia not included). Follow up data are available for 755 of 984 (77%) lesions and constitute the basis of this evaluation. Follow-up mammograms were performed of 728 lesions at 6 to 67 months (mean 24, median 17.8) after VB. RESULTS: Seven technically unsuccessful cases underwent immediate rebiopsy; 3 unsuccessful cases were diagnosed otherwise. No false negative occurred among the 752 followed-up, eventually successful VBs. On follow-up mammography 444 of 728 (61%) benign lesions proved radiologically completely removed, 284 (39%) partially. In 6 cases (0.8%) a surgical biopsy was performed again during the follow-up time confirming the benign result. No scar was seen in 96%, a slight scar in 3.8%, and a small stellate scar with possible diagnostic interference in 0.3%. CONCLUSIONS: A benign diagnosis of quality assured VB is very reliable and leads to no or minimal scarring. PMID- 12885606 TI - Influence of follow-up methodology and completeness on apparent clinical outcome of fundoplication. AB - BACKGROUND: Outcomes after surgery can be determined by face-to-face interview, interviewing patients by telephone, or by mailing out questionnaires. For reasons of convenience, many clinical research studies use mail survey methods. Mail-out responses, however, are rarely complete, and patients returning mail-out questionnaires might be different from patients who do not return questionnaires. METHODS: We identified a group of 75 patients who were all sent a mail-out questionnaire 5 or more years after a laparoscopic fundoplication. Some of these patients returned questionnaires and some did not. All patients were subsequently interviewed by telephone. The responses to the two different data collection methods were compared as well as the interview responses of patients who returned questionnaires with those of patients who did not to determine the influence of follow-up methodology on apparent clinical outcome. RESULTS: Of the 75 patients interviewed, 49 patients (65%) had previously returned a mail-out questionnaire and 26 patients had not. The mean time difference between mail-out response and telephone interview was 1 month. When mail-out responses were compared with interview responses of the patients who had returned questionnaires, equivalent answers were obtained for all symptoms except for dysphagia, for which a higher incidence and greater scores were reported by patients in their mail-out questionnaire responses. When mail-out nonresponders were compared with responders, the nonresponders had a lower level of overall satisfaction with the outcome of surgery, a higher incidence of dysphagia, and greater levels of heartburn. CONCLUSIONS: The measurement of surgical outcomes using a mail-out questionnaire may not be an accurate method of assessing patient symptoms and satisfaction after surgery, particularly if follow-up is incomplete. In this study, patients not responding to mail-out questionnaires were more likely to have adverse symptoms and a lower level of satisfaction than patients returning questionnaires. This dissatisfaction could make patients less likely to return mail-out questionnaires, and for this reason studies with incomplete follow-up that rely on mail-out questionnaire responses may report erroneously higher surgical success rates. PMID- 12885607 TI - Postherpetic self-limited abdominal wall herniation. PMID- 12885608 TI - Survival benefits of portal vein resection for pancreatic cancer. AB - BACKGROUND: The efficacy of portal vein resection for pancreatic cancer is controversial. METHODS: Eighty-one consecutive patients with pancreatic cancer undergoing surgical resection were retrospectively analyzed. The clinicopathological findings and relationship between portal vein resection and survival were investigated. RESULTS: Thirty-three patients with pancreatic cancer underwent pancreatic resection with portal vein resection. Histological examination revealed that 17 patients had definite invasion to the portal vein (group 1) and 16 patients had no invasion (group 2). Forty-eight patients with pancreatic cancer underwent pancreatic resection without portal vein resection (group 3). There were no significant differences in survival rates (P = 0.437) between patients with portal vein resection and patients without portal vein resection. However, patients in group 1 had a significantly (P = 0.021) worse prognosis as compared with those in group 2. Despite aggressive surgical resection, the surgical margin was positive in 35% of patients in group 1 as compared with 13% of patients in group 2 and 21% of patients in group 3. CONCLUSIONS: Patients undergoing portal vein resection for pancreatic cancer had a prognosis similar to patients without portal vein resection. Negative microscopic invasion to the portal vein was significantly associated with improved survival. PMID- 12885609 TI - Impact of socioeconomic deprivation and primary pathology on rate of reversal of Hartmann's procedure. AB - BACKGROUND: Hartmann's procedure is an operation for left-sided colonic pathology where primary anastomosis is considered unsafe. The aim of this study was to assess the impact of socioeconomic deprivation and primary pathology on the rate of reanastomosis. METHODS: All patients who underwent Hartmann's procedure between 1992 and 2000 in our unit were included. The deprivation category (DEPCAT) scores from 1 (affluent) to 7 (most deprived) were calculated. RESULTS: An emergency procedure was performed in 118 of 124 patients. Primary pathology was malignant in 40 and benign in 84 patients. The DEPCAT scores were high (6,7) in 76 and intermediate (3 to 5) in 42 patients. Of the 102 patients surviving the postoperative period, only 23 underwent reversal. The reversal rate was lower in patients with malignancy (P = 0.005) and higher DEPCAT scores. CONCLUSIONS: Primary diagnosis and socioeconomic deprivation adversely influence the rate of reversal of Hartmann's procedure. PMID- 12885610 TI - Endoscopic components separation for abdominal compartment syndrome. AB - BACKGROUND: Sustained intraabdominal pressures of 14 to 20 mm Hg have significant pathophysiological consequences, but there is currently no satisfactory low morbidity procedure appropriate for intervention early in the disease process of abdominal compartment syndrome (ACS). The anatomical principles of abdominal wall components separation were used to develop a percutaneous procedure that increased abdominal capacity and decreased abdominal pressure. METHODS: Using a porcine model, we determined abdominal capacity changes by helium insufflation. Corn oil was then used to create an episode of sustained intraabdominal hypertension and changes in intraabdominal pressure and intestinal mucosal oxygenation were determined. RESULTS: Endoscopic abdominal wall components separation (EACS) increased abdominal capacity by 1 L (from 0.89 +/- 0.39 L to 1.95 +/- 0.48 L; P <0.001). During intraabdominal hypertension, EACS decreased abdominal pressure by 31.6% (from 15.9 +/- 2.1 to 11.0 +/- 1.5 mm Hg; P <0.001). Intestinal PO(2) was increased by 61% (18.8 +/- 11.4 to 30.3 +/- 11.7; P = 0.012) CONCLUSIONS: A minimally invasive procedure (EACS) is feasible and has demonstrated effectiveness in a porcine model of ACS. PMID- 12885611 TI - How we do a bloodless partial splenectomy. AB - Partial splenectomy and Tru-cut biopsy are not routinely practiced because of the lack of vascular control to arrest bleeding. Using radiofrequency energy to coagulate the resection margin and biopsy tract, a 74-year-old woman with a tumor in the lower pole of the spleen underwent partial splenectomy and Tru-Cut biopsy of the spleen. Hemostasis was excellent. Blood loss was minimal and the patient was discharged with a functioning spleen. This new technique may allow safe and bloodless partial splenic resection and Tru-cut biopsy of the spleen, which might reduce the number of splenectomies performed and the consequent difficulties for the patient that can arise. PMID- 12885612 TI - Venous thrombosis. PMID- 12885614 TI - Do you know your students' basic clinical skills exposure? AB - BACKGROUND: Undergraduate medical education in the United States is changing. Many medical schools have developed a set of basic clinical skills (BCS) that all students are required to have mastered; however, very few have acquired objective information regarding specific student experiences. The purpose of this study was to determine the BCS encounters for junior medical students at a large midwestern university utilizing a handheld personal digital assistant (PDA). METHODS: A core curriculum of BCS was proposed and involved 52 procedures/skills. An electronic BCS database was developed utilizing HanDBase software and then placed on a PDA (Palm) and distributed to 25 third-year medical students randomly as they entered their clinical year. Students logged their skill encounters for 9 months and then electronically transferred the database by e-mail. RESULTS: Students participated in 1,115 procedural/skill encounters (range 17 to 90; median 41; average 44.6). Of the 52 core BCS, all students performed 10. Fewer than 50% of students had any exposure to very common skills. Thirty-four percent of skill encounters occurred at a county hospital, 19% at a clinic, 10% at a university hospital, 10% at a private hospital, 7% at a VA hospital, 4% at a children's hospital, and 16% at miscellaneous locations. CONCLUSIONS: The PDA devices were simple and convenient to use, while allowing for easy transfer and tabulation of database information by electronic mail. Significant gaps in BCS exposure were noted across the curriculum. Mentor sign-off on the PDA permitted early feedback opportunities. We can now begin to reward educators for skills mentoring and perform formal assessment of BCS within specific clerkships to enhance future educational objectives. PMID- 12885613 TI - Effect of sleep deprivation on the performance of simulated laparoscopic surgical skill. AB - BACKGROUND: Resident work hours may impact patient care. We hypothesized that "call-associated" acute sleep deprivation has no effect on technical dexterity as measured on a minimally invasive surgery trainer, virtual reality (MIST VR) surgical simulator. METHODS: Thirty-five surgical residents were prospectively evaluated pre-call (rested), on-call (rested), and post-call (acutely sleep deprived). Participants completed questionnaires regarding sleep hours and level of fatigue. Technical skill was assessed using the MIST VR. Speed, errors, and economy of motion were automatically recorded by the MIST VR computer simulator. Data were analyzed by paired Student t test and analysis of variance. RESULTS: Estimated hours of sleep and subjective indicators of fatigue were different between rested and sleep-deprived residents. The number of errors and time to complete all tasks increased at the post-call assessment. CONCLUSIONS: Resident work schedules lead to sleep deprivation and fatigue. Call-associated sleep deprivation and fatigue are associated with increased technical errors in the performance of simulated laparoscopic surgical skills. PMID- 12885615 TI - A study of career choice patterns among Canadian medical students. AB - BACKGROUND: The number of students pursuing general surgery (GS) has declined in Canada. The reasons for this, and program directors' (PDs) perception of it, are unclear. METHODS: A survey was distributed to medical students at Queen's University and the University of Manitoba, and to all Canadian GS program directors to explore causes for the trend and identify potential solutions. RESULTS: Students pursuing GS were more likely to feel that GS fit their lifestyle needs (P < 0.05) and to have met positive role models (P < 0.05). Hardship of the training and practice, length of training, need for prestige, income and enjoyment of procedures did not correlate with specialty choice. Half of GS PDs did not perceive a decline in the number of applicants. CONCLUSIONS: Attention to student-raised concerns regarding GS programs and increased awareness of the applicants' trends among PDs can be used to address the current decline in applications to GS. PMID- 12885616 TI - Impact of an endovascular program on the operative experience of abdominal aortic aneurysm in vascular fellowship and general surgery residency. AB - BACKGROUND: This study was performed to determine the impact of an endovascular program (EVP) on open and endovascular abdominal aortic aneurysm (AAA) operations in a residency training institution. METHODS: Over an 8-year period ending in September 2001, hospital records of all patients undergoing open or endovascular AAA repair were retrospectively reviewed. Data were analyzed to determine the changing patterns of case volume, type of operative repair, and complexity of open repair with regards to the training of both general surgical chief residents and vascular fellows. RESULTS: A total of 849 AAA operations were performed during the study period. The initiation of the EVP in 1997 resulted in a steady increase in the total annual AAA cases (P < 0.05), due in part to an increase in endovascular AAA operations despite a decrease in the annual open AAA volume. EVP had a positive impact on the overall operative experience of vascular fellows owing to the large increase in their endovascular AAA experience (annual mean pre EVP 3 +/- 0.8 versus post-EVP 47 +/- 9.6, P < 0.01). A significant reduction occurred in the vascular fellows' open AAA experience (annual mean pre-EVP 40 +/- 12.7 versus post-EVP 19 +/- 9.4, P < 0.05). EVP did not affect the endovascular AAA experience of general surgery chief residents (annual mean pre-EVP 1 +/- 0.8 versus post-EVP 3 +/- 1.5, not significant). A significant reduction occurred in chief residents' open AAA experience (annual mean pre-EVP 39 +/- 9.7 versus post EVP 18 +/- 7.4, P < 0.05). EVP did not affect the operative experience of complex open AAA operations in either vascular fellows or general surgery residents. CONCLUSIONS: An endovascular program has a positive impact on the aortic aneurysm practice in an academic institution, as evidenced by the significant increase in annual endovascular AAA cases despite a decrease in open AAA operations. Although vascular fellows continued to maintain sufficient experience in both open and endovascular AAA operations, general surgery chief residents suffered a significant decrease in their open AAA experience. Further evaluation of the residency system is warranted to better optimize the training paradigm of both vascular fellowship and general surgery residency. PMID- 12885617 TI - Geographic variation in posttreatment surveillance intensity for patients with cutaneous melanoma. AB - BACKGROUND: We investigated whether geographic determinants could account for variation in posttreatment melanoma surveillance intensity among plastic surgeons. METHODS: A custom-designed questionnaire was mailed to U.S. and non U.S. members of the American Society of Plastic and Reconstructive Surgeons (ASPRS). Subjects were asked how they use 14 specific follow-up modalities during years 1 to 5 and 10 following primary treatment for patients with cutaneous melanoma. Repeated-measures analysis of variance was used to compare practice patterns by TNM stage, year postsurgery, U.S. census region, metropolitan statistical area (MSA), and managed care organization (MCO) penetration rate. RESULTS: Of the 1,142 respondents, 395 were evaluable. Those who did not perform melanoma surgery or follow-up were excluded. Correlation analysis showed that mean follow-up intensity for the modalities surveyed was highly correlated across TNM stages and years postsurgery. Within MSAs, only chest radiograph utilization varied significantly. The pattern of testing varied significantly by geographic region for seven modalities (office visit, computed tomography scan of the brain and chest/abdomen, alpha-fetoprotein level, 5S-cysteinyl dopa level, abdominal ultrasonogram, bone scan); in each of these, utilization by non-U.S. surgeons exceeded utilization in any U.S. census region. The pattern of testing varied significantly by MCO penetration rate for chest radiograph (greater utilization in the lowest MCO penetration rate areas) and 5S-cysteinyl dopa level (greater utilization in the highest MCO penetration rate areas). CONCLUSIONS: The intensity of posttreatment surveillance recommended by ASPRS members caring for patients with cutaneous melanoma varies markedly. This analysis provides the first evidence that geographic factors significantly affect the surveillance strategies of clinicians following patients with cutaneous melanoma. Variation by census region was most prominent, although the size of detected differences was small. PMID- 12885618 TI - Sutureless mesh-plug femoral hernioplasty. AB - BACKGROUND: Prosthetic mesh groin hernioplasty is well established. We aimed to determine whether a sutureless mesh-plug repair of femoral hernias was a reliable proposition. METHODS: A prospective study of 42 patients undergoing femoral hernioplasty with a preformed three-dimensional mesh plug placed in the preperitoneal plane. RESULTS: There were no complications, and no recurrences were detected at a median follow-up of 2 years (range 2 to 44). CONCLUSIONS: Preperitoneal mesh plug femoral hernioplasty appears to be universally applicable and deserves further evaluation. PMID- 12885619 TI - Molecular dynamics decomposition of temperature-dependent elastic neutron scattering by a protein solution. AB - Molecular dynamics simulations are performed of bovine pancreatic trypsin inhibitor in a cryosolution over a range of temperatures from 80 to 300 K and the origins identified of elastic dynamic neutron scattering from the solution. The elastic scattering and mean-square displacement calculated from the molecular dynamics trajectories are in reasonable agreement with experiments on a larger protein in the same solvent. The solvent and protein contributions to the scattering from the simulation model are determined. At lower temperatures (< approximately 200 K) or on shorter timescales ( approximately 10 ps) the scattering contributions are proportional to the isotopic nuclear scattering cross-sections of each component. However, for T > 200 K marked deviations from these cross-sections are seen due to differences in the dynamics of the components of the solution. Rapid activation of solvent diffusion leads to the variation with temperature of the total elastic intensity being determined largely by that of the solvent. At higher temperatures (>240 K) and longer times ( approximately 100 ps) the protein makes the only significant contribution to the scattering, the solvent scattering having moved out of the accessible time space window. Decomposition of the protein mean-square displacement shows that the observed dynamical transition in the solution at 200-220 K involves activation of both internal motions and external whole-molecule rotational and translational diffusion. The proportion that the external dynamics contributes to the protein mean-square displacement increases to approximately 30 and 60% at 300 K on the 10- and 100-ps timescales, respectively. PMID- 12885620 TI - Specific ion effects: why the properties of lysozyme in salt solutions follow a Hofmeister series. AB - Protein solubility in aqueous solutions depends in a complicated and not well understood way on pH, salt type, and salt concentration. Why for instance does the use of two different monovalent salts, potassium thiocyanate and potassium chloride, produce such different results? One important and previously neglected source of ion specificity is the ionic dispersion potential that acts between each ion and the protein. This attractive potential is found to be much stronger for SCN(-) than it is for Cl(-). We present model calculations, performed within a modified ion-specific double-layer theory, that demonstrate the large effect of including these ionic dispersion potentials. The results are consistent with experiments performed on hen egg-white lysozymes and on neutral black lipid membranes. The calculated surface pH and net lysozyme charge depend strongly on the choice of anion. We demonstrate that the lysozyme net charge is larger, and the corresponding Debye length shorter, in a thiocyanate salt solution than in a chloride salt solution. Recent experiments have suggested that pK(a) values of histidines depend on salt concentration and on ionic species. We finally demonstrate that once ionic dispersion potentials are included in the theory these results can quantitatively be reinterpreted in terms of a highly specific surface pH (and a salt-independent pK(a)). PMID- 12885621 TI - The unbinding of ATP from F1-ATPase. AB - Using molecular dynamics, we study the unbinding of ATP in F(1)-ATPase from its tight binding state to its weak binding state. The calculations are made feasible through use of interpolated atomic structures from Wang and Oster [Nature 1998, 396: 279-282]. These structures are applied to atoms distant from the catalytic site. The forces from these distant atoms gradually drive a large primary region through a series of sixteen equilibrated steps that trace the hinge bending conformational change in the beta-subunit that drives rotation of gamma-subunit. As the rotation progresses, we find a sequential weakening and breaking of the hydrogen bonds between the ATP molecule and the alpha- and beta-subunits of the ATPase. This finding agrees with the "binding-zipper" model [Oster and Wang, BIOCHIM: Biophys. Acta 2000, 1458: 482-510.] In this model, the progressive formation of the hydrogen bonds is the energy source driving the rotation of the gamma-shaft during hydrolysis. Conversely, the corresponding sequential breaking of these bonds is driven by rotation of the shaft during ATP synthesis. Our results for the energetics during rotation suggest that the nucleotide's coordination with Mg(2+) during binding and release is necessary to account for the observed high efficiency of the motor. PMID- 12885622 TI - Theoretical modeling of prion disease incubation. AB - We apply a theoretical aggregation model to laboratory and epidemiological prion disease incubation time data. In our model, slow growth of misfolded protein aggregates from small initial seeds controls the latent or lag phase; aggregate fissioning and subsequent spreading leads to an exponential growth phase. Our model accounts for the striking reproducibility of incubation times for high dose inoculation of lab animals. In particular, low dose yields broad incubation time distributions, and increasing dose narrows distributions and yields sharply defined onset times. We also explore how incubation time statistics depend upon aggregate morphology. We apply our model to fit the experimental dose-incubation curves for distinct strains of scrapie, and explain logarithmic variation at high dose and deviations from logarithmic behavior at low dose. We use this to make testable predictions for infectivity time-course experiments. PMID- 12885623 TI - Effective conductivity of a suspension of permeabilized cells: a theoretical analysis. AB - During the electroporation cell membrane undergoes structural changes, which increase the membrane conductivity and consequently lead to a change in effective conductivity of a cell suspension. To correlate microscopic membrane changes to macroscopic changes in conductivity of a suspension, we analyzed the effective conductivity theoretically, using two different approaches: numerically, using the finite elements method; and analytically, by using the equivalence principle. We derived the equation, which connects membrane conductivity with effective conductivity of the cell suspension. The changes in effective conductivity were analyzed for different parameters: cell volume fraction, membrane and medium conductivity, critical transmembrane potential, and cell orientation. In our analysis we used a tensor form of the effective conductivity, thus taking into account the anisotropic nature of the cell electropermeabilization and rotation of the cells. To determine the effect of cell rotation, as questioned by some authors, the difference between conductivity of a cell suspension with normally distributed orientations and parallel orientation was also calculated, and determined to be <10%. The presented theory provides a theoretical basis for the analysis of measurements of the effective conductivity during electroporation. PMID- 12885625 TI - Metabolic switching in the sugar phosphotransferase system of Escherichia coli. AB - Bacteria grown in a mixture of multiple sugars will first metabolize a preferred sugar until it is nearly depleted, only then turning to other carbon sources in the medium. This sharp switching of metabolic preference is characteristic of systems that optimize fitness. Here we consider the mechanism by which switching can occur in the Escherichia coli phosphotransferase system (PTS), which regulates the uptake and metabolism of several sugars. Using a model combining the description of fast biochemical processes and slower genetic regulation, we derive metabolic phase diagrams for the uptake of two PTS sugars, indicating regions of distinct sugar preference as a function of external sugar concentrations. We then propose a classification of bacterial phenotypes based on the topology of the metabolic phase diagram, and enumerate the possible topologically distinct phenotypes that can be achieved through mutations of the PTS. This procedure reveals that there is only one nontrivial switching phenotype that is insensitive to large changes in biochemical parameters. This phenotype exhibits diauxic growth, a manifestation of the winner-take-all dynamics enforced by PTS architecture. Winner-take-all behavior is implemented by the induction of sugar-specific operons, combined with competition between sugars for limited phosphoryl flux. We propose that flux-limited competition could be a common mechanism for introducing repressive interactions in cellular networks, and we argue that switching behavior similar to that described here should occur generically in systems that implement such a mechanism. PMID- 12885624 TI - An integrated model of epidermal growth factor receptor trafficking and signal transduction. AB - Endocytic trafficking of many types of receptors can have profound effects on subsequent signaling events. Quantitative models of these processes, however, have usually considered trafficking and signaling independently. Here, we present an integrated model of both the trafficking and signaling pathway of the epidermal growth factor receptor (EGFR) using a probability weighted-dynamic Monte Carlo simulation. Our model consists of hundreds of distinct endocytic compartments and approximately 13,000 reactions/events that occur over a broad spatio-temporal range. By using a realistic multicompartment model, we can investigate the distribution of the receptors among cellular compartments as well as their potential signal transduction characteristics. Our new model also allows the incorporation of physiochemical aspects of ligand-receptor interactions, such as pH-dependent binding in different endosomal compartments. To determine the utility of this approach, we simulated the differential activation of the EGFR by two of its ligands, epidermal growth factor (EGF) and transforming growth factor alpha (TGF-alpha). Our simulations predict that when EGFR is activated with TGF alpha, receptor activation is biased toward the cell surface whereas EGF produces a signaling bias toward the endosomal compartment. Experiments confirm these predictions from our model and simulations. Our model accurately predicts the kinetics and extent of receptor downregulation induced by either EGF or TGF alpha. Our results suggest that receptor trafficking controls the compartmental bias of signal transduction, rather than simply modulating signal magnitude. Our model provides a new approach to evaluating the complex effect of receptor trafficking on signal transduction. Importantly, the stochastic and compartmental nature of the simulation allows these models to be directly tested by high throughput approaches, such as quantitative image analysis. PMID- 12885626 TI - Ribosome recycling, diffusion, and mRNA loop formation in translational regulation. AB - We explore and quantify the physical and biochemical mechanisms that may be relevant in the regulation of translation. After elongation and detachment from the 3' termination site of mRNA, parts of the ribosome machinery can diffuse back to the initiation site, especially if it is held nearby, enhancing overall translation rates. The elongation steps of the mRNA-bound ribosomes are modeled using exact and asymptotic results of the totally asymmetric exclusion process. Since the ribosome injection rates of the totally asymmetric exclusion process depend on the local concentrations at the initiation site, a source of ribosomes emanating from the termination end can feed back to the initiation site, leading to a self-consistent set of equations for the steady-state ribosome throughput. Additional mRNA binding factors can also promote loop formation, or cyclization, bringing the initiation and termination sites into close proximity. The probability distribution of the distance between the initiation and termination sites is described using simple noninteracting polymer models. We find that the initiation, or initial ribosome adsorption binding required for maximal throughput, can vary dramatically depending on certain values of the bulk ribosome concentration and diffusion constant. If cooperative interactions among the loop-promoting proteins and the initiation/termination sites are considered, the throughput can be further regulated in a nonmonotonic manner. Experiments that can potentially test the hypothesized physical mechanisms are discussed. PMID- 12885627 TI - Lateral heterogeneity of photosystems in thylakoid membranes studied by Brownian dynamics simulations. AB - The aggregation and segregation of photosystems in higher plant thylakoid membranes as stromal cation-induced phenomena are studied by the Brownian dynamics method. A theoretical model of photosystems lateral movement within the membrane plane is developed, assuming their pairwise effective potential interaction in aqueous and lipid media and their diffusion. Along with the screened electrostatic repulsive interaction the model accounts for the van der Waals-type, elastic, and lipid-induced attractive forces between photosystems of different sizes and charges. Simulations with a priori estimated parameters demonstrate that all three studied repulsion-attraction alternatives might favor the local segregation of photosystems under physiologically reasonable conditions. However, only the lipid-induced potential combined with the size corrected screened Coulomb interaction provides the segregated configurations with photosystems II localized in the central part of the grana-size simulation cell and photosystems I occupying its margins, as observed experimentally. Mapping of thermodynamic states reveals that the coexistence curves between isotropic and aggregated phases are the sigmoidlike functions regardless of the effective potential type. It correlates with measurements of the chlorophyll content of thylakoid fragments. Also the universality of the phase curves characterizes the aggregation and segregation of photosystems as order-disorder phase transitions with the Debye radius as a governing parameter. PMID- 12885628 TI - Insights into nucleic acid conformational dynamics from massively parallel stochastic simulations. AB - The helical hairpin is one of the most ubiquitous and elementary secondary structural motifs in nucleic acids, capable of serving functional roles and participating in long-range tertiary contacts. Yet the self-assembly of these structures has not been well-characterized at the atomic level. With this in mind, the dynamics of nucleic acid hairpin formation and disruption have been studied using a novel computational tool: large-scale, parallel, atomistic molecular dynamics simulation employing an inhomogeneous distributed computer consisting of more than 40,000 processors. Using multiple methodologies, over 500 micro s of atomistic simulation time has been collected for a large ensemble of hairpins (sequence 5'-GGGC[GCAA]GCCU-3'), allowing characterization of rare events not previously observable in simulation. From uncoupled ensemble dynamics simulations in unperturbed folding conditions, we report on 1), competing pathways between the folded and unfolded regions of the conformational space; 2), observed nonnative stacking and basepairing traps; and 3), a helix unwinding rewinding mode that is differentiated from the unfolding and folding dynamics. A heterogeneous transition state ensemble is characterized structurally through calculations of conformer-specific folding probabilities and a multiplexed replica exchange stochastic dynamics algorithm is used to derive an approximate folding landscape. A comparison between the observed folding mechanism and that of a peptide beta-hairpin analog suggests that although native topology defines the character of the folding landscape, the statistical weighting of potential folding pathways is determined by the chemical nature of the polymer. PMID- 12885629 TI - Effect of DNA superhelicity and bound proteins on mechanistic aspects of the Hin mediated and Fis-enhanced inversion. AB - Using a recently developed inhomogeneous, macroscopic model for long DNA bound to proteins, we examine topological and geometric aspects of DNA/protein structures and dynamics on various stages of the Hin inversion pathway. This biological reaction involves exchange of DNA in a synaptic complex that brings together several DNA sites bound to Hin dimers as well as Fis enhancers. Brownian dynamics simulations in the millisecond timescale allow us to follow and analyze the DNA/protein dynamics trajectories and to examine the effects of DNA superhelicity and protein binding on various reaction steps. Analysis of the generated kinetic pathways helps explain mechanistic aspects regarding the process by which two or three protein-bound DNA sites come to close spatial proximity and show that how topological selectivity (two trapped supercoils), enhancer binding, and properties of supercoiled DNA play critical roles in regulating the inversion reaction. Specifically, a critical amount of DNA superhelicity (e.g., |sigma| > 0.02) leads to an optimal interplay for the first reaction step-two-site juxtaposition-between large-scale random rearrangements of Hin-bound DNA and local slithering within branches of plectonemes. The three-site juxtaposition, the second step, is significantly accelerated by the presence of an enhancer protein that, due to severe local bending, also alters juxtaposition mechanisms, especially for superhelical density magnitude greater than around 0.04. PMID- 12885630 TI - Instabilities in the transient response of muscle. AB - We investigate the isometric transient response of muscle using a quantitative stochastic model of the actomyosin cycle based on the swinging lever-arm hypothesis. We first consider a single pair of filaments, and show that when values of parameters such as the lever-arm displacement and the cross-bridge elasticity are chosen to provide effective energy transduction, the T(2) curve (the tension recovered immediately after a step displacement) displays a region of negative slope. If filament compliance and the discrete nature of the binding sites are taken into account, the negative slope is diminished, but not eliminated. This implies that there is an instability in the dynamics of individual half sarcomeres. However, when the symmetric nature of whole sarcomeres is taken into account, filament rearrangement becomes important during the transient: as tension is recovered, some half sarcomeres lengthen whereas others shorten. This leads to a flat T(2) curve, as observed experimentally. In addition, we investigate the isotonic transient response and show that for a range of parameter values the model displays damped oscillations, as recently observed in experiments on single muscle fibers. We conclude that it is essential to consider the collective dynamics of many sarcomeres, rather than the dynamics of a single pair of filaments, when interpreting the transient response of muscle. PMID- 12885631 TI - The motility of mollicutes. AB - Recent experiments show that the conformation of filament proteins play a role in the motility and morphology of many different types of bacteria. Conformational changes in the protein subunits may produce forces to drive propulsion and cell division. Here we present a molecular mechanism by which these forces can drive cell motion. Coupling of a biochemical cycle, such as ATP hydrolysis, to the dynamics of elastic filaments enable elastic filaments to propagate deformations that generate propulsive forces. We demonstrate this possibility for two classes of wall-less bacteria called mollicutes: the swimming of helical-shaped Spiroplasma, and the gliding motility of Mycoplasma. PMID- 12885632 TI - Helix rotation model of the flagellar rotary motor. AB - A new model of the flagellar motor is proposed that is based on established dynamics of the KcsA potassium ion channel and on known genetic, biochemical, and biophysical facts, which accounts for the mechanics of torque generation, force transmission, and reversals of motor rotation. It predicts that proton (or in some species sodium ion) flow generates short, reversible helix rotations of the MotA-MotB channel complex (the stator) that are transmitted by Coulomb forces to the FliG segments at the rotor surface. Channels are arranged as symmetric pairs, S and T, that swing back and forth in synchrony. S and T alternate in attaching to the rotor, so that force transmission proceeds in steps. The sense of motor rotation can be readily reversed by conformationally switching the position of charged groups on the rotor so that they interact with the stator during the reverse rather than forward strokes. An elastic device accounts for the observed smoothness of rotation and a prolonged attachment of the torque generators to the rotor, i.e., a high duty ratio of each torque-generating unit. PMID- 12885633 TI - Structure-based prediction of potential binding and nonbinding peptides to HIV-1 protease. AB - HIV-1 protease is a major drug target against AIDS as it permits viral maturation by processing the gag and pol polyproteins of the virus. The cleavage sites in these polyproteins do not have obvious sequence homology or a binding motif and the specificity of the protease is not easily determined. We used various threading approaches, together with the crystal structures of substrate complexes which served as template structures, to study the substrate specificity of HIV-1 protease with the aim of obtaining a better differentiation between binding and nonbinding sequences. The predictions from threading improved when distance dependent interaction energy functions were used instead of contact matrices. To rank the peptides and properly account for the peptide's conformation in the total energy, the results from using short-range potentials on multiple template structures were averaged. Finally, a dynamic threading approach is introduced which is potentially useful for cases when there is only one template structure available. The conformational energy of the peptide-especially the term accounting for the side chains-was found to be important in differentiating between binding and nonbinding sequences. Hence, the substrate specificity, and thus the ability of the virus to mature, is affected by the compatibility of the substrate peptide to fit within the limited conformational space of the active site groove. PMID- 12885634 TI - A computer simulation study of the hydrated proton in a synthetic proton channel. AB - Classical molecular dynamics simulations using the multistate empirical valence bond model for aqueous proton transport were performed to characterize the hydration structure of an excess proton inside a leucine-serine synthetic ion channel, LS2. For such a nonuniform pore size ion channel, it is found that the Zundel ion (H(5)O(2)(+)) solvation structure is generally more stable in narrow channel regions than in wider channel regions, which is in agreement with a recent study on idealized hydrophobic proton channels. However, considerable diversity in the relative stability of the Zundel to Eigen cation (H(9)O(4)(+)) was observed. Three of the five wide channel regions, one located at the channel's center and the other two located near the channel mouths, are found to show extraordinary preference for the Eigen solvation structure. This implies that proton hopping is inhibited in these regions and therefore suggests that these regions may behave as barriers in the proton conducting pathway inside the channel. The proton solvation is also greatly influenced by the local molecular environment of the protein. In particular, the polar side chains of the Ser residues, which are intimately involved in the solvation structure, can greatly influence proton solvation. However, no preference of the influence by the various Ser side chains was found; they can either promote or prevent the formation of certain solvation structures. PMID- 12885635 TI - CymA of Klebsiella oxytoca outer membrane: binding of cyclodextrins and study of the current noise of the open channel. AB - CymA, the outer membrane component of the cyclodextrin (CD) uptake and metabolism system of Klebsiella oxytoca, was reconstituted into lipid bilayer membranes. The channel properties of this unusual porin were studied in detail. The binding of CDs to the channel resulted in its complete block for ion transport. This result allowed the detailed investigation of carbohydrate binding, and the stability constants for the binding of cyclic and linear carbohydrates to the binding site inside the channel were calculated from titration experiments of the membrane conductance with the carbohydrates. Highest stability constant was observed for alpha-cyclodextrin (alpha-CD; K = 32,000 1/M) followed by beta-cyclodextrin (beta CD; K = 1970 1/M) and gamma-cyclodextrin (gamma-CD; K = 310 1/M). Linear maltooligosaccharides bound also to CymA but with much smaller stability constants as compared to cyclic ones. The noise of the current through CymA in multi- and single-channel experiments was investigated using fast Fourier transformation. The current through the open channels had a rather high spectral density, which was a Lorentzian function of the frequency up to 2000 Hz. Upon addition of cyclic dextrins to the aqueous phase the spectral density decreased in a dose-dependent manner, which made it impossible to evaluate the binding kinetics. Experiments with single CymA-channels demonstrated the channel is highly asymmetric concerning channel flickers and current noise. PMID- 12885636 TI - The role of small intraprotein cavities in the catalytic cycle of bacteriorhodopsin. AB - The last phase of the proton transfer cycle of bacteriorhodopsin calls for a passage of a proton from D38 to D96. This reaction utilizes a narrow shaft approximately 10-A long that connects the two carboxylates that cross through a very hydrophobic domain. As the shaft is too narrow to be permanently hydrated, there are two alternatives for the proton propagation into the channel. The proton may propagate through the shaft without solvation at the expense of a high electrostatic barrier; alternatively, the shaft will expand to accommodate some water molecules, thus lowering the Born energy for the insertion of the charge into the protein (B. Schatzler, N. A. Dencher, J. Tittor, D. Oesterhelt, S. Yaniv Checover, E. Nachliel, and G. Gutman, 2003, BIOPHYS: J. 84:671-686). A comparative study of nine published crystal-structures of bacteriorhodopsin identified, next to the shaft, microcavities in the protein whose position and surrounding atoms are common to the reported structures. Some of the cavities either shrink or expand during the photocycle. It is argued that the plasticity of the cavities provides a working space needed for the transient solvation of the shaft, thus reducing the activation energy necessary for the solvation of the shaft. This suggestion is corroborated by the recent observations of Klink et al. (B. U. Klink, R. Winter, M. Engelhard, and I. Chizhov, 2002, BIOPHYS: J. 83:3490 3498) that the late phases of the photocycle (tau >/=1 ms) are strongly inhibited by external pressure. PMID- 12885637 TI - Partitioning of individual flexible polymers into a nanoscopic protein pore. AB - Polymer dynamics are of fundamental importance in materials science, biotechnology, and medicine. However, very little is known about the kinetics of partitioning of flexible polymer molecules into pores of nanometer dimensions. We employed electrical recording to probe the partitioning of single poly(ethylene glycol) (PEG) molecules, at concentrations near the dilute regime, into the transmembrane beta-barrel of individual protein pores formed from staphylococcal alpha-hemolysin (alphaHL). The interactions of the alpha-hemolysin pore with the PEGs (M(w) 940-6000 Da) fell into two classes: short-duration events (tau approximately 20 micro s), approximately 85% of the total, and long-duration events (tau approximately 100 micro s), approximately 15% of the total. The association rate constants (k(on)) for both classes of events were strongly dependent on polymer mass, and values of k(on) ranged over two orders of magnitude. By contrast, the dissociation rate constants (k(off)) exhibited a weak dependence on mass, suggesting that the polymer chains are largely compacted before they enter the pore, and do not decompact to a significant extent before they exit. The values of k(on) and k(off) were used to determine partition coefficients (Pi) for the PEGs between the bulk aqueous phase and the pore lumen. The low values of Pi are in keeping with a negligible interaction between the PEG chains and the interior surface of the pore, which is independent of ionic strength. For the long events, values of Pi decrease exponentially with polymer mass, according to the scaling law of Daoud and de Gennes. For PEG molecules larger than approximately 5 kDa, Pi reached a limiting value suggesting that these PEG chains cannot fit entirely into the beta-barrel. PMID- 12885638 TI - Tryptophan scanning of D1S6 and D4S6 C-termini in voltage-gated sodium channels. AB - Recent reports suggest that four S6 C-termini may jointly close the voltage-gated cation channel at the cytoplasmic side, probably as an inverted teepee structure. In this study we substituted individually a total of 18 residues at D1S6 and D4S6 C-terminal ends of the rNav1.4 Na(+) channel alpha-subunit with tryptophan (W) and examined their corresponding gating properties when expressed in Hek293t cells along with beta1 subunit. Several W-mutants displayed significant changes in activation, fast inactivation, and/or slow inactivation gating. In particular, five S6 W-mutants showed incomplete fast inactivation with noninactivating maintained currents present. Cysteine (C) substitutions of these five residues resulted in two mutants with slightly more maintained currents. Multiple substitutions at these five positions yielded two mutants (L437C/A438W, L435W/L437C/A438W) that exhibited phenotypes with minimal fast inactivation. Unexpectedly, such inactivation-deficient mutants expressed Na(+) currents as well as did the wild-type. Furthermore, all mutants with impaired fast inactivation exhibited an enhanced slow inactivation phenotype. Implications of these results will be discussed in terms of indirect allosteric modulations via amino acid substitutions and/or a direct involvement of S6 C-termini in Na(+) channel gating. PMID- 12885639 TI - Calmodulin modulates initiation but not termination of spontaneous Ca2+ sparks in frog skeletal muscle. AB - Calmodulin is a ubiquitous Ca(2+) sensing protein that binds to and modulates the sarcoplasmic reticulum Ca(2+) release channel, ryanodine receptor (RYR). Here we assessed the effects of calmodulin on the local Ca(2+) release properties of RYR in permeabilized frog skeletal muscle fibers. Fluorescently labeled recombinant calmodulin in the internal solution localized at the Z-line/triad region. Calmodulin (0.05-5.0 micro M) in the internal solution (free [Ca(2+)](i) approximately 50-100 nM) initiated a highly cooperative dose-dependent increase in Ca(2+) spark frequency, with a half-maximal activation (K) of 1.1 micro M, a Hill coefficient (n) of 4.2 and a fractional maximal increase in frequency (R) of 17-fold. A non-Ca(2+) binding mutant of calmodulin elicited a similar highly cooperative dose-dependent increase in spark frequency (K = 1.0 micro M; n = 3.7; R = 12-fold). Spatiotemporal properties of Ca(2+) sparks were essentially unaffected by either wild-type or mutant calmodulin. An N-terminal extension of calmodulin, (N+3)calmodulin, that binds to but does not activate RYR at nM [Ca(2+)] in sarcoplasmic reticulum vesicles, prevented the calmodulin-induced increase in spark frequency. These data suggest that exogenous Ca(2+)-free calmodulin cooperatively sensitizes the Ca(2+) release channel to open, but that Ca(2+) binding to the added calmodulin does not play a significant role in the termination of Ca(2+) sparks. PMID- 12885640 TI - Regulation of calcium channel activity by lipid domain formation in planar lipid bilayers. AB - The sarcoplasmic reticulum channel (ryanodine receptor) from cardiac myocytes was reconstituted into planar lipid bilayers consisting of 1-palmitoyl-2-oleoyl phosphatidylethanolamine (POPE) and 1-palmitoyl-2-oleoyl-phosphatidylcholine (POPC) in varying ratios. The channel activity parameters, i.e., open probability and average open time and its resolved short and long components, were determined as a function of POPE mole fraction (X(PE)) at 22.4 degrees C. Interestingly, all of these parameters exhibited a narrow and pronounced peak at X(PE) approximately 0.80. Differential scanning calorimetric measurements on POPE/POPC liposomes with increasing X(PE) indicated that the lipid bilayer enters a composition-driven transition from the liquid-crystalline state to the gel state at 22.4 degrees C when X(PE) approaches 0.80. Thus, the peaking of the reconstituted channel activity at X(PE) approximately 0.80 in the planar bilayer could result from the appearance of gel/liquid-crystalline domain boundaries at this POPE content. Lipid packing at domain boundaries is known to be looser as compared to the homogenous gel or liquid-crystalline state. We propose that the attractive potential of packing defects at lipid domain boundaries and entropic excluded volume effects could result in the direct interactions of the transmembrane region of the channel protein with the lipid-packing defects at the lipid/protein interface, which could thus provide a favorable environment for the open state of the protein. The present findings indicate that the activity of the sarcoplasmic reticulum calcium channel could be modulated by lipid domain formation upon slight changes in membrane lipid composition in vivo. PMID- 12885641 TI - Alpha-bungarotoxin binding to acetylcholine receptor membranes studied by low angle X-ray diffraction. AB - The nicotinic acetylcholine receptor (nAChR) carries two binding sites for snake venom neurotoxins. alpha-Bungarotoxin from the Southeast Asian banded krait, Bungarus multicinctus, is a long neurotoxin which competitively blocks the nAChR at the acetylcholine binding sites in a relatively irreversible manner. Low angle x-ray diffraction was used to generate electron density profile structures at 14 A resolution for Torpedo californica nAChR membranes in the absence and presence of alpha-bungarotoxin. Analysis of the lamellar diffraction data indicated a 452 A lattice spacing between stacked nAChR membrane pairs. In the presence of alpha bungarotoxin, the quality of the diffraction data and the lamellar lattice spacing were unchanged. In the plane of the membrane, the nAChRs packed together with a nearest neighbor distance of 80 A, and this distance increased to 85 A in the presence of toxin. Electron density profile structures were calculated in the absence and presence of alpha-bungarotoxin, revealing a location for the toxin binding sites. In native, fully-hydrated nAChR membranes, alpha-bungarotoxin binds to the nAChR outer vestibule and contacts the surface of the membrane bilayer. PMID- 12885642 TI - Multistep mechanism of chloride translocation in a strongly anion-selective porin channel. AB - The strongly anion-selective porin channel Omp32 from the bacterium Delftia acidovorans differs from other unspecific porins by its pronounced selectivity for anions and its particularly small channel cross-section. Multinanosecond molecular dynamics simulations of chloride ion movement in this pore protein suggest that translocated anions interact intimately with the charges of a "basic ladder", whose dynamics lead the anions in a stepwise manner through the constriction zone of the channel. The ladder-steps comprise the central clustered arginine groups and flanking basic residues at its exoplasmic and periplasmic sides. The computed free energy profile of ion movement in and around the constriction zone shows a corresponding succession of free energy minima and barriers. A number of polar atoms from other amino acids contribute to the coordination of Cl(-) at certain sites and to its temporary immobilization in the channel. A special binding site occurs at the transition of the constriction zone to the periplasmic funnel, binding the chloride ion over significant lengths of time. The results from our MD study offer a possible explanation for the nonlinear conductance properties and unusual salt-dependent characteristics of Omp32 observed earlier in experimental measurements. PMID- 12885643 TI - ATP-independent luminal oscillations and release of Ca2+ and H+ from mast cell secretory granules: implications for signal transduction. AB - InsP(3) is an important link in the intracellular information network. Previous observations show that activation of InsP(3)-receptor channels on the granular membrane can turn secretory granules into Ca(2+) oscillators that deliver periodic trains of Ca(2+) release to the cytosol (T. Nguyen, W. C. Chin, and P. Verdugo, 1998, Nature, 395:908-912; I. Quesada, W. C. Chin, J. Steed, P. Campos Bedolla, and P. Verdugo, 2001, BIOPHYS: J. 80:2133-2139). Here we show that InsP(3) can also turn mast cell granules into proton oscillators. InsP(3)-induced intralumenal [H(+)] oscillations are ATP-independent, result from H(+)/K(+) exchange in the heparin matrix, and produce perigranular pH oscillations with the same frequency. These perigranular pH oscillations are in-phase with intralumenal [H(+)] but out-of-phase with the corresponding perigranular [Ca(2+)] oscillations. The low pH of the secretory compartment has critical implications in a broad range of intracellular processes. However, the association of proton release with InsP(3)-induced Ca(2+) signals, their similar periodic nature, and the sensitivity of important exocytic proteins to the joint action of Ca(2+) and pH strongly suggests that granules might encode a combined Ca(2+)/H(+) intracellular signal. A H(+)/Ca(2+) signal could significantly increase the specificity of the information sent by the granule by transmitting two frequency encoded messages targeted exclusively to proteins like calmodulin, annexins, or syncollin that are crucial for exocytosis and require specific combinations of [Ca(2+)] "and" pH for their action. PMID- 12885644 TI - Interaction of viscotoxins A3 and B with membrane model systems: implications to their mechanism of action. AB - Viscotoxins are small proteins that are thought to interact with biomembranes, displaying different toxic activities against a varied number of cell types, being viscotoxin A(3) (VtA(3)) the most cytotoxic whereas viscotoxin B (VtB) is the less potent. By using infrared and fluorescence spectroscopies, we have studied the interaction of VtA(3) and VtB, both wild and reduced ones, with model membranes containing negatively charged phospholipids. Both VtA(3) and VtB present a high conformational stability, and a similar conformation both in solution and when bound to membranes. In solution, the infrared spectra of the reduced proteins show an increase in bandwidth compared to the nonreduced ones indicating a greater flexibility. VtA(3) and VtB bind with high affinity to membranes containing negatively charged phospholipids and are motional restricted, their binding being dependent on phospholipid composition. Whereas nonreduced proteins maintain their structure when bound to membranes, reduced ones aggregate. Furthermore, leakage experiments show that wild proteins were capable of disrupting membranes whereas reduced proteins were not. The effect of VtA(3) and VtB on membranes having different phospholipid composition is diverse, affecting the cooperativity and fluidity of the membranes. Viscotoxins interact with membranes in a complex way, most likely organizing themselves at the surface inducing the appearance of defects that lead to the destabilization and disruption of the membrane bilayer. PMID- 12885645 TI - Charge-dependent translocation of the Trojan peptide penetratin across lipid membranes. AB - We studied the interaction of the cell-penetrating peptide penetratin with mixed dioleoylphosphatidylcholine/dioleoylphoshatidylglycerol (DOPC/DOPG) unilamellar vesicles as a function of the molar fraction of anionic lipid, X(PG), by means of isothermal titration calorimetry. The work was aimed at getting a better understanding of factors that affect the peptide binding to lipid membranes and its permeation through the bilayer. The binding was well described by a surface partitioning equilibrium using an effective charge of the peptide of z(P) approximately 5.1 +/- 0.5. The peptide first binds to the outer surface of the vesicles, the effective binding capacity of which increases with X(PG). At X(PG) approximately 0.5 and a molar ratio of bound peptide-to-lipid of approximately 1/20 the membranes become permeable and penetratin binds also to the inner monolayer after internalization. The results were rationalized in terms of an "electroporation-like" mechanism, according to which the asymmetrical distribution of the peptide between the outer and inner surfaces of the charged bilayer causes a transmembrane electrical field, which alters the lateral and the curvature stress acting within the membrane. At a threshold value these effects induce internalization of penetratin presumably via inversely curved transient structures. PMID- 12885646 TI - The pressure-dependence of the size of extruded vesicles. AB - Variations in the size of vesicles formed by extrusion through small pores are discussed in terms of a simple model. Our model predicts that the radius should decrease as the square root of the applied pressure, consistent with data for vesicles extruded under various conditions. The model also predicts dependencies on the pore size used and on the lysis tension of the vesicles being extruded that are consistent with our data. The pore size was varied by using track-etched polycarbonate membranes with average pore diameters ranging from 50 to 200 nm. To vary the lysis tension, vesicles made from POPC (1-palmitoyl-2-oleoyl-sn-glycero 3-phosphatidylcholine), mixtures of POPC and cholesterol, and mixtures of POPC and C(16)-ceramide were studied. The lysis tension, as measured by an extrusion based technique, of POPC:cholesterol vesicles is higher than that of pure POPC vesicles whereas POPC:ceramide vesicles have lower lysis tensions than POPC vesicles. PMID- 12885647 TI - Oxygen permeation profile in lipid membranes: comparison with transmembrane polarity profile. AB - Permeation of oxygen into membranes is relevant not only to physiological function, but also to depth determinations in membranes by site-directed spin labeling. Spin-lattice (T(1)) relaxation enhancements by air or molecular oxygen were determined for phosphatidylcholines spin labeled at positions (n = 4-14, 16) of the sn-2 chain in fluid membranes of dimyristoyl phosphatidylcholine, by using nonlinear continuous-wave electron paramagnetic resonance (EPR). Both progressive saturation and out-of-phase continuous-wave EPR measurements yield similar oxygen permeation profiles. With pure oxygen, the T(2)-relaxation enhancements determined from homogeneous linewidths of the linear EPR spectra are equal to the T(1)-relaxation enhancements determined by nonlinear EPR. This confirms that both relaxation enhancements occur by Heisenberg exchange, which requires direct contact between oxygen and spin label. Oxygen concentrates in the hydrophobic interior of phospholipid bilayer membranes with a sigmoidal permeation profile that is the inverse of the polarity profile established earlier for these spin labeled lipids. The shape of the oxygen permeation profile in fluid lipid membranes is controlled partly by the penetration of water, via the transmembrane polarity profile. At the protein interface of the KcsA ion channel, the oxygen profile is more diffuse than that in fluid lipid bilayers. PMID- 12885648 TI - Hydration and lateral organization in phospholipid bilayers containing sphingomyelin: a 2H-NMR study. AB - Interfacial properties of lipid bilayers were studied by (2)H nuclear magnetic resonance spectroscopy, with emphasis on a comparison between phosphatidylcholine and sphingomyelin. Spectral resolution and sensitivity was improved by macroscopic membrane alignment. The motionally averaged quadrupolar interaction of interlamellar deuterium oxide was employed to probe the interfacial polarity of the membranes. The D(2)O quadrupolar splittings indicated that the sphingomyelin lipid-water interface is less polar above the phase transition temperature T(m) than below T(m). The opposite behavior was found in phosphatidylcholine bilayers. Macroscopically aligned sphingomyelin bilayers also furnished (2)H-signals from the amide residue and from the hydroxyl group of the sphingosine moiety. The rate of water-hydroxyl deuteron exchange could be measured, whereas the exchange of the amide deuteron was too slow for the inversion-transfer technique employed, suggesting that the amide residue is involved in intermolecular hydrogen bonding. Order parameter profiles in mixtures of sphingomyelin and chain-perdeuterated phosphatidylcholine revealed an ordering effect as a result of the highly saturated chains of the sphingolipids. The temperature dependence of the (2)H quadrupolar splittings was indicative of lateral phase separation in the mixed systems. The results are discussed with regard to interfacial structure and lateral organization in sphingomyelin containing biomembranes. PMID- 12885649 TI - Lipid membrane polarity profiles by high-field EPR. AB - Profiles of polarity across biological membranes are essential determinants of the cellular permeability barrier and of the stability of transmembrane proteins. High-field electron paramagnetic resonance of systematically spin-labeled lipid chains is used here to determine the polarity profiles of cholesterol-containing phospholipid membranes. The polarity dependence of the g(xx)-tensor element is opposite to the dependence on chain dynamics, and additionally has enhanced sensitivity to hydrogen bonding. Both features make high-field measurements superior to conventional determinations of local polarity from spin-label hyperfine couplings. The profile of g(xx) in dimyristoyl phosphatidylcholine membranes with 5 or 40 mol% cholesterol is established with eleven positional isomers of phosphatidylcholine, spin labeled at positions n = 4-14 in the sn-2 chain. A sigmoidal barrier, centered about chain position n(o) approximately 8, mirrors the corresponding sigmoidal trough obtained from the spin-label hyperfine coupling, A(zz). For the different positions, n, it is found that partial differential g(xx)/ partial differential A(zz) = -2.4 T(-1), a high value that is characteristic of hydrogen-bonded spin labels. This demonstrates that the transmembrane polarity profile registered by spin labels corresponds to water penetration into the membrane. Inhomogeneous broadening of the g(xx)-spectral feature demonstrates heterogeneities of the water distribution in the regions of higher intramembrane polarity defined by n < 8. In the transition region between high- and low-polarity regions (n approximately 8), the g(xx)-feature consists of two components characteristic of coexisting hydrated and nonhydrated states. PMID- 12885650 TI - Fluorescence energy transfer reveals microdomain formation at physiological temperatures in lipid mixtures modeling the outer leaflet of the plasma membrane. AB - An approach is described using fluorescence resonance energy transfer (FRET) to detect inhomogeneity in lipid organization, on distance scales of the order of tens of nanometers or greater, in lipid bilayers. This approach compares the efficiency of energy transfer between two matched fluorescent lipid donors, differing in their affinities for ordered versus disordered regions of the bilayer, and an acceptor lipid that distributes preferentially into disordered regions. Inhomogeneities in bilayer organization, on spatial scales of tens of nanometers or greater, are detected as a marked difference in the efficiencies of quenching of fluorescence of the two donor species by the acceptor. Using a novel pair of 7-nitrobenz-2-oxa-1,3-diazol-4-yl (NBD)-labeled tetraacyl lipids as donor species with a rhodaminyl-labeled acceptor, this strategy faithfully reports homo versus inhomogeneous mixing in each of several lipid bilayer systems whose organization on the FRET distance scale can be predicted from previous findings. Interestingly, however, the present FRET method reports clear evidence of inhomogeneity in the organization of mixtures combining sphingomyelin or saturated phospholipids with unsaturated phospholipids and physiological proportions of cholesterol, even at physiological temperatures where these systems have been reported to appear homogeneous by fluorescence microscopy. These results indicate that under physiological conditions, lipid mixtures mimicking the lipid composition of the outer leaflet of the plasma membrane can form domains on a spatial scale comparable to that inferred for the dimensions of lipid rafts in biological membranes. PMID- 12885652 TI - Ionic interactions play a role in the regulatory mechanism of scallop heavy meromyosin. AB - Heavy meromyosin from scallop (scHMM) striated muscle is regulated by calcium binding to the essential light chain. The regulation can be modeled with a calcium-dependent equilibrium between on and off scHMM conformations. The observed rate constant for mant-ADP dissociation from scHMM is calcium dependent, and we show here that it can be used to define the equilibrium constant (K(eq)) between on and off conformations. The data show that K(eq) is markedly ionic strength dependent, with high salt (>/=200 mM) abolishing the off state even in the absence of calcium and low salt (<50 mM) favoring the off state even in the presence of calcium. Debye-Huckel plots of the equilibrium constant (K(eq)) for the on and off forms gave parallel slopes (5.94 +/- 0.33 and 6.36 +/- 0.17 M( 0.5)) in the presence and absence of calcium. The presence of an equilibrium mixture of two conformations was confirmed by sedimentation data and the effects of ADP, calcium and ionic strength were in qualitative agreement. Thus scHMM exists in two conformations that can be distinguished in sedimentation profiles and by the rate of release of mant-ADP. Increasing salt concentrations biases the system toward the on state, suggesting a role for ionic interactions in stabilizing the off state. PMID- 12885651 TI - Regulatory proteins alter nucleotide binding to acto-myosin of sliding filaments in motility assays. AB - The sliding speed of unregulated thin filaments in motility assays is only about half that of the unloaded shortening velocity of muscle fibers. The addition of regulatory proteins, troponin and tropomyosin, is known to increase the sliding speed of thin filaments in the in vitro motility assay. To learn if this effect is related to the rate of MgADP dissociation from the acto-S1 cross-bridge head, the effects of regulatory proteins on nucleotide binding and release in motility assays were measured in the presence and absence of regulatory proteins. The apparent affinity of acto-heavy meromyosin (acto-HMM) for MgATP was reduced by the presence of regulatory proteins. Similarly, the regulatory proteins increase the concentration of MgADP required to inhibit sliding. These results suggest that regulatory proteins either accelerate the rate of MgADP release from acto HMM-MgADP or slow its binding to acto-HMM. The reduction of temperature also altered the relationship between thin filament sliding speed and the regulatory proteins. At lower temperatures, the regulatory proteins lost their ability to increase thin filament sliding speed above that of unregulated thin filaments. It is hypothesized that structural changes in the actin portion of the acto-myosin interface are induced by regulatory protein binding to actin. PMID- 12885653 TI - Myosin head configuration in relaxed insect flight muscle: x-ray modeled resting cross-bridges in a pre-powerstroke state are poised for actin binding. AB - Low-angle x-ray diffraction patterns from relaxed insect flight muscle recorded on the BioCAT beamline at the Argonne APS have been modeled to 6.5 nm resolution (R-factor 9.7%, 65 reflections) using the known myosin head atomic coordinates, a hinge between the motor (catalytic) domain and the light chain-binding (neck) region (lever arm), together with a simulated annealing procedure. The best head conformation angles around the hinge gave a head shape that was close to that typical of relaxed M*ADP*Pi heads, a head shape never before demonstrated in intact muscle. The best packing constrained the eight heads per crown within a compact crown shelf projecting at approximately 90 degrees to the filament axis. The two heads of each myosin molecule assume nonequivalent positions, one head projecting outward while the other curves round the thick filament surface to nose against the proximal neck of the projecting head of the neighboring molecule. The projecting heads immediately suggest a possible cross-bridge cycle. The relaxed projecting head, oriented almost as needed for actin attachment, will attach, then release Pi followed by ADP, as the lever arm with a purely axial change in tilt drives approximately 10 nm of actin filament sliding on the way to the nucleotide-free limit of its working stroke. The overall arrangement appears well designed to support precision cycling for the myogenic oscillatory mode of contraction with its enhanced stretch-activation response used in flight by insects equipped with asynchronous fibrillar flight muscles. PMID- 12885654 TI - Comparison between the predictions of diffusion-reaction models and localized Ca2+ transients in amphibian skeletal muscle fibers. AB - We developed a three-dimensional cylindrical diffusion-reaction model of a single amphibian myofibril in which Ca(2+) release occurred only at the Z-line. The model incorporated diffusion of Ca(2+), Mg(2+), and all relevant buffer species, as well as the kinetic binding reactions between the buffers and appropriate ions. Model data was blurred according to a Gaussian approximation of the point spread function of the microscope and directly compared with experimental data obtained using the confocal spot methodology. The flux parameters were adjusted until the simulated Z-line transient matched the experimental one. This model could not simultaneously predict key parameters of the experimental M- and Z-line transients, even when model parameters were adjusted to unreasonably extreme values. Even though the model was accurate in predicting the Z-line transient under conditions of high [EGTA], it predicted a significantly narrower Ca(2+) domain than observed experimentally. We modified the model to incorporate a broader band of release centered at the Z-line. This extended release model was superior both in simultaneously predicting critical features of the Z- and M-line transients as well as the domain profile under conditions of high [EGTA]. We conclude that a model of release occurring exclusively at the Z-line cannot explain our experimental data and suggest that Ca(2+) may be released from a broader region of the sarcoplasmic reticulum than just the T-tubule-sarcoplasmic reticulum junction. PMID- 12885655 TI - The conformation of myosin head domains in rigor muscle determined by X-ray interference. AB - In the absence of adenosine triphosphate, the head domains of myosin cross bridges in muscle bind to actin filaments in a rigor conformation that is expected to mimic that following the working stroke during active contraction. We used x-ray interference between the two head arrays in opposite halves of each myosin filament to determine the rigor head conformation in single fibers from frog skeletal muscle. During isometric contraction (force T(0)), the interference effect splits the M3 x-ray reflection from the axial repeat of the heads into two peaks with relative intensity (higher angle/lower angle peak) 0.76. In demembranated fibers in rigor at low force (<0.05 T(0)), the relative intensity was 4.0, showing that the center of mass of the heads had moved 4.5 nm closer to the midpoint of the myosin filament. When rigor fibers were stretched, increasing the force to 0.55 T(0), the heads' center of mass moved back by 1.1-1.6 nm. These motions can be explained by tilting of the light chain domain of the head so that the mean angle between the Cys(707)-Lys(843) vector and the filament axis increases by approximately 36 degrees between isometric contraction and low-force rigor, and decreases by 7-10 degrees when the rigor fiber is stretched to 0.55 T(0). PMID- 12885656 TI - Solvation of nucleosides in aqueous mixtures of organic solvents: relevance to DNA open basepairs. AB - Toward the goal of understanding how open basepairs in DNA interact with their heterogeneous environment, we have studied the steady-state intrinsic fluorescence properties of the purine and pyrimidine deoxynucleosides in organic solvents in the presence of small amounts of water. The organic solvents used in the present study were: n-butanol, acetonitrile, methanol, n-propanol, isopropanol, and isobutanol. For n-butanol and acetonitrile, which have a high degree of amphiphilicity and weak hydrogen bonding ability, respectively, the fluorescence spectral properties of the purines are found to depend on the sequence of steps in which the aqueous mixtures were formed. By contrast, no such dependence was observed in the mixtures with any of the other solvents used in the present study. Moreover, no such dependence was observed for the pyrimidines. These findings suggest that the final solvation network around the purines is dependent on the nature of the environment to which they were initially exposed. This would tend to present an impediment to the closing of AT or GC basepairs in DNA that become open as a result of structural fluctuations, DNA bending, or protein-DNA interactions. PMID- 12885657 TI - Direct measurement of the photoelectric response time of bacteriorhodopsin via electro-optic sampling. AB - The photovoltaic signal associated with the primary photochemical event in an oriented bacteriorhodopsin film is measured by directly probing the electric field in the bacteriorhodopsin film using an ultrafast electro-optic sampling technique. The inherent response time is limited only by the laser pulse width of 500 fs, and permits a measurement of the photovoltage with a bandwidth of better than 350 GHz. All previous published studies have been carried out with bandwidths of 50 GHz or lower. We observe a charge buildup with an exponential formation time of 1.68 +/- 0.05 ps and an initial decay time of 31.7 ps. Deconvolution with a 500-fs Gaussian excitation pulse reduces the exponential formation time to 1.61 +/- 0.04 ps. The photovoltaic signal continues to rise for 4.5 ps after excitation, and the voltage profile corresponds well with the population dynamics of the K state. The origin of the fast photovoltage is assigned to the partial isomerization of the chromophore and the coupled motion of the Arg-82 residue during the primary event. PMID- 12885658 TI - A general model for amyloid fibril assembly based on morphological studies using atomic force microscopy. AB - Based on atomic force microscopy analysis of the morphology of fibrillar species formed during fibrillation of alpha-synuclein, insulin, and the B1 domain of protein G, a previously described model for the assembly of amyloid fibrils of immunoglobulin light-chain variable domains is proposed as a general model for the assembly of protein fibrils. For all of the proteins studied, we observed two or three fibrillar species that vary in diameter. The smallest, protofilaments, have a uniform height, whereas the larger species, protofibrils and fibrils, have morphologies that are indicative of multiple protofilaments intertwining. In all cases, protofilaments intertwine to form protofibrils, and protofibrils intertwine to form fibrils. We propose that the hierarchical assembly model describes a general mechanism of assembly for all amyloid fibrils. PMID- 12885659 TI - TOUCHSTONE II: a new approach to ab initio protein structure prediction. AB - We have developed a new combined approach for ab initio protein structure prediction. The protein conformation is described as a lattice chain connecting C(alpha) atoms, with attached C(beta) atoms and side-chain centers of mass. The model force field includes various short-range and long-range knowledge-based potentials derived from a statistical analysis of the regularities of protein structures. The combination of these energy terms is optimized through the maximization of correlation for 30 x 60,000 decoys between the root mean square deviation (RMSD) to native and energies, as well as the energy gap between native and the decoy ensemble. To accelerate the conformational search, a newly developed parallel hyperbolic sampling algorithm with a composite movement set is used in the Monte Carlo simulation processes. We exploit this strategy to successfully fold 41/100 small proteins (36 approximately 120 residues) with predicted structures having a RMSD from native below 6.5 A in the top five cluster centroids. To fold larger-size proteins as well as to improve the folding yield of small proteins, we incorporate into the basic force field side-chain contact predictions from our threading program PROSPECTOR where homologous proteins were excluded from the data base. With these threading-based restraints, the program can fold 83/125 test proteins (36 approximately 174 residues) with structures having a RMSD to native below 6.5 A in the top five cluster centroids. This shows the significant improvement of folding by using predicted tertiary restraints, especially when the accuracy of side-chain contact prediction is >20%. For native fold selection, we introduce quantities dependent on the cluster density and the combination of energy and free energy, which show a higher discriminative power to select the native structure than the previously used cluster energy or cluster size, and which can be used in native structure identification in blind simulations. These procedures are readily automated and are being implemented on a genomic scale. PMID- 12885660 TI - 3D structure of Sulfolobus solfataricus carboxypeptidase developed by molecular modeling is confirmed by site-directed mutagenesis and small angle X-ray scattering. AB - Sulfolobus solfataricus carboxypeptidase (CPSso) is a thermostable zinc metalloenzyme with a M(r) of 43,000. Taking into account the experimentally determined zinc content of one ion per subunit, we developed two alternative 3D models, starting from the available structures of Thermoactinomyces vulgaris carboxypeptidase (Model A) and Pseudomonas carboxypeptidase G2 (Model B). The former enzyme is monomeric and has one metal ion in the active site, while the latter is dimeric and has two bound zinc ions. The two models were computed by exploiting the structural alignment of the one zinc- with the two zinc-containing active sites of the two templates, and with a threading procedure. Both computed structures resembled the respective template, with only one bound zinc with tetrahedric coordination in the active site. With these models, two different quaternary structures can be modeled: one using Model A with a hexameric symmetry, the other from Model B with a tetrameric symmetry. Mutagenesis experiments directed toward the residues putatively involved in metal chelation in either of the models disproved Model A and supported Model B, in which the metal-binding site comprises His(108), Asp(109), and His(168). We also identified Glu(142) as the acidic residue interacting with the water molecule occupying the fourth chelation site. Furthermore, the overall fold and the oligomeric structure of the molecule was validated by small angle x-ray scattering (SAXS). An ab initio original approach was used to reconstruct the shape of the CPSso in solution from the experimental curves. The results clearly support a tetrameric structure. The Monte Carlo method was then used to compare the crystallographic coordinates of the possible quaternary structures for CPSso with the SAXS profiles. The fitting procedure showed that only the model built using the Pseudomonas carboxypeptidase G2 structure as a template fitted the experimental data. PMID- 12885661 TI - Molecular dynamics simulation of dimeric and monomeric forms of human prion protein: insight into dynamics and properties. AB - A central theme in prion protein research is the detection of the process that underlies the conformational transition from the normal cellular prion form (PrP(C)) to its pathogenic isoform (PrP(Sc)). Although the three-dimensional structures of monomeric and dimeric human prion protein (HuPrP) have been revealed by NMR spectroscopy and x-ray crystallography, the process underlying the conformational change from PrP(C) to PrP(Sc) and the dynamics and functions of PrP(C) remain unknown. The dimeric form is thought to play an important role in the conformational transition. In this study, we performed molecular dynamics (MD) simulations on monomeric and dimeric HuPrP at 300 K and 500 K for 10 ns to investigate the differences in the properties of the monomer and the dimer from the perspective of dynamic and structural behaviors. Simulations were also undertaken with Asp178Asn and acidic pH, which is known as a disease-associated factor. Our results indicate that the dynamics of the dimer and monomer were similar (e.g., denaturation of helices and elongation of the beta-sheet). However, additional secondary structure elements formed in the dimer might result in showing the differences in dynamics and properties between the monomer and dimer (e.g., the greater retention of dimeric than monomeric tertiary structure). PMID- 12885662 TI - Concentration-dependent tetramerization of bovine visual arrestin. AB - The oligomeric states of bovine visual arrestin in solution were studied by small angle x-ray scattering. The Guinier plot of arrestin at the concentration ranging from 0.4 mg/ml to 11.1 mg/ml was approximated with a straight line, and the apparent molecular weight was evaluated by the concentration-normalized intensity at zero angle (I(0)/conc). Using ovalbumin as a molecular weight standard, it was found that arrestin varied from monomer to tetramer depending on the concentration. The I(0)/conc decreased at high-salt concentration, but was independent of temperature. The simulation analysis of the concentration dependent increase of I(0)/conc demonstrated that the tetramerization is highly cooperative, and arrestin at the physiological concentration is virtually in the equilibrium between monomer and tetramer. The concentration of arrestin monomer, which is considered to be an active form, remains at an almost constant level even if the total concentration of arrestin fluctuates within the physiological range. The scattering profile of arrestin tetramer in solution was in good agreement with that in the crystal, indicating that the quaternary structure in solution is essentially identical to that in crystal. Small-angle x-ray scattering was applied to a binding assay of phosphorylated rhodopsin and arrestin in the detergent system, and we directly observed their association as the increase of I(0)/conc. PMID- 12885663 TI - Conformation, orientation, and adsorption kinetics of dermaseptin B2 onto synthetic supports at aqueous/solid interface. AB - The antimicrobial activity of cationic amphipathic peptides is due mainly to the adsorption of peptides onto target membranes, which can be modulated by such physicochemical parameters as charge and hydrophobicity. We investigated the structure of dermaseptin B2 (Drs B2) at the aqueous/synthetic solid support interface and its adsorption kinetics using attenuated total reflection Fourier transform infrared spectroscopy and surface plasmon resonance. We determined the conformation and affinity of Drs B2 adsorbed onto negatively charged (silica or dextran) and hydrophobic supports. Synthetic supports of differing hydrophobicity were obtained by modifying silica or gold with omega-functionalized alkylsilanes (bromo, vinyl, phenyl, methyl) or alkylthiols. The peptide molecules adsorbed onto negatively charged supports mostly had a beta-type conformation. In contrast, a monolayer of Drs B2, mainly in the alpha-helical conformation, was adsorbed irreversibly onto the hydrophobic synthetic supports. The conformational changes during formation of the adsorbed monolayer were monitored by two dimensional Fourier transform infrared spectroscopy correlation; they showed the influence of peptide-peptide interactions on alpha-helix folding on the most hydrophobic support. The orientation of the alpha-helical Drs B2 with respect to the hydrophobic support was determined by polarized attenuated total reflection; it was around 15 +/- 5 degrees. This orientation was confirmed and illustrated by a molecular dynamics study. These combined data demonstrate that specific chemical environments influence the structure of Drs B2, which could explain the many functions of antimicrobial peptides. PMID- 12885664 TI - Calculation of mutational free energy changes in transition states for protein folding. AB - Recent advances in experimental and computational methods have made it possible to determine with considerable accuracy the structures whose formation is rate limiting for the folding of some small proteins-the transition state ensemble, or TSE. We present a method to analyze and validate all-atom models of such structures. The method is based on the comparison of experimental data with the computation of the change in free energy of the TSE resulting from specific mutations. Each mutation is modeled individually in all members of an ensemble of transition state structures using a method originally developed to predict mutational changes in the stability of native proteins. We first apply this method to six proteins for which we have determined the TSEs with a technique that uses experimental mutational data (Phi-values) as restraints in the structure determination and find a highly significant correlation between the calculated free energy changes and those derived from experimental kinetic data. We then use the procedure to analyze transition state structures determined by molecular dynamics simulations of unfolding, again finding a high correlation. Finally, we use the method to estimate changes in folding rates of several hydrophobic core mutants of Fyn SH3. Taken together, these results show that the procedure developed here is a tool of general validity for analyzing, assessing, and improving the quality of the structures of transition states for protein folding. PMID- 12885665 TI - Proline can have opposite effects on fast and slow protein folding phases. AB - Proline isomerization is well known to cause additional slow phases during protein refolding. We address a new question: does the presence of prolines significantly affect the very fast kinetics that lead to the formation of folding intermediates? We examined both the very slow (10-100 min) and very fast (4 micro s-2.5 ms) folding kinetics of the two-domain enzyme yeast phosphoglycerate kinase by temperature-jump relaxation. Phosphoglycerate kinase contains a conserved cis proline in position 204, in addition to several trans-prolines. Native cis prolines have the largest effect on folding kinetics because the unfolded state favors trans isomerization, so we compared the kinetics of a P204H mutant with the wild-type as a proof of principle. The presence of Pro-204 causes an additional slow phase upon refolding from the cold denatured state, as reported in the literature. Contrary to this, the fast folding events are sped up in the presence of the cis-proline, probably by restriction of the conformational space accessible to the molecule. The wild-type and Pro204His mutant would be excellent models for off-lattice simulations probing the effects of conformational restriction on short timescales. PMID- 12885666 TI - The shape parameter of liposomes and DNA-lipid complexes determined by viscometry utilizing small sample volumes. AB - A minicapillary viscometer utilizing <0.5 ml of sample at a volume fraction of <0.1% is described. The calculated a/b of DPPC/DPPG multilamellar liposome was 1.14 as prolate ellipsoids and a/b of dioleoylpropyltrimethyl ammonium methylsulfate-DNA complex at a charge ratio of 4:1 (+/-) was 3.7 as prolate ellipsoids or 4.9 as oblate ellipsoids. The deviation of shape from perfect sphere is thus expressed quantitatively in more than two significant figures. In these measurement, the necessary amount of DNA is <0.5 mg. PMID- 12885667 TI - Atomic force microscopy and light scattering of small unilamellar actin containing liposomes. AB - Three-dimensional networks of filamentous actin (F-actin) encapsulated inside phosphatidylcholine liposomes are currently being used in an effort to model the cytoskeleton and plasma membrane of eukaryotic cells. In this article, unilamellar lipid vesicles consisting of egg yolk-derived phosphatidylcholine encapsulating monomeric actin (G-actin) were made via extrusion in low ionic strength buffer (G-buffer). Vesicle shape and structure in these dispersions was studied using a combination of fluid-tapping atomic force microscopy, and multiangle static light scattering. After subjecting the liposome dispersion to high ionic strength polymerization buffer (F-buffer) containing K(+) ions, atomic force microscopy imaging and light scattering of these liposomes indicated the formation of specialized structures, including an overall liposome structure transformation from spherical to torus, disk-shaped geometries and tubular assemblies. Several atomic force microscopy control measurements were made to ascertain that the specialized structures formed were not due to free G-actin and F-actin self-assembling on the sample surface, plain liposomes exposed to G- and F-buffer, or liposomes encapsulating G-actin. Liposomes encapsulating G-actin assumed mostly thin disk shapes and some large irregularly shaped aggregates. In contrast, liposomes encapsulating polymerized actin assumed mostly torus or disk shapes along with some high aspect ratio tubular structures. PMID- 12885668 TI - Effects of a carane derivative local anesthetic on a phospholipid bilayer studied by molecular dynamics simulation. AB - Molecular dynamics (MD) simulations of two hydrated palmitoyloleoylphosphatidylcholine (POPC) bilayers each containing eight carane derivative (KP-23) local anesthetic (LA) molecules in neutral (POPC-LA) or protonated (POPC-LAH) forms were carried out to investigate the effect of KP-23 and its protonation on the bilayer. 3-ns trajectories were used for analyses. A pure POPC bilayer was employed as a reference system. In both POPC-LA and POPC LAH systems a few KP-23 molecules intercalated into the bilayer and moved near the bilayer/water interface. They were located on the hydrophobic core side of the interface in the POPC-LA bilayer, but on the water phase side in the POPC-LAH bilayer. The order of the POPC chains was higher in the POPC-LA bilayer than in the pure POPC bilayer and was lower in the POPC-LAH bilayer. Interactions between polar groups of KP-23 and POPC or water were responsible for a lower hydration of POPC headgroups in POPC bilayers containing KP-23 than in the pure POPC bilayer. KP-23 molecules were found to form aggregates both in POPC-LA and POPC-LAH bilayers. Due to higher amphiphilicity of LAH, the LAH aggregate was more micelle like and larger than the LA one. The results demonstrate the rapid timescales of the initial processes that take place at and near the bilayer interface as well as details of the atomic level interactions between local anesthetic and the lipid matrix of a cell membrane. PMID- 12885669 TI - Small angle X-ray scattering study of meso-tetrakis (4-sulfonatophenyl) porphyrin in aqueous solution: a self-aggregation model. AB - The aggregate morphology of meso-tetrakis(4-sulfonatophenyl) porphyrin (TPPS(4)) in aqueous solution is investigated by using small angle x-ray scattering (SAXS) technique. Measurements were performed at pH 4.0 and 9.0 to monitor the pH influence on the structural parameters of the aggregates. Radii of gyration were obtained from distance distribution functions p(r) analysis. The experimental data of TPPS(4) at pH 4.0 showed well-defined oscillations characteristic of large aggregates in contrast to the SAXS curve of 5 mM TPPS(4) at pH 9.0, where both a significant decrease in the intensity and the disappearance of the oscillation peaks suggest the dissociation of the aggregate. A 340-A long "hollow" cylinder with shell thickness of 20 A, compatible to the porphyrin molecule dimension, represents well the scattering curve of the aggregates at pH 4.0. According to the fitting parameters, 26 porphyrin molecules self-associate into a ringlike configuration in the plane of the cylinder cross-section. The total number of porphyrin molecules in the whole aggregate was also estimated as approximately 3000. The model compatible to SAXS data of a hollow cylinder with J aggregation in the cross-section and H-aggregation (columnar stacking) between the cylinder layers is consistent with optical absorption spectroscopic data both in the literature and obtained in this work. PMID- 12885670 TI - Protein flexibility and conformational state: a comparison of collective vibrational modes of wild-type and D96N bacteriorhodopsin. AB - Far infrared (FIR) spectral measurements of wild-type (WT) and D96N mutant bacteriorhodopsin thin films have been carried out using terahertz time domain spectroscopy as a function of hydration, temperature, and conformational state. The results are compared to calculated spectra generated via normal mode analyses using CHARMM. We find that the FIR absorbance is slowly increasing with frequency and without strong narrow features over the range of 2-60 cm(-1) and up to a resolution of 0.17 cm(-1). The broad absorption shifts in frequency with decreasing temperature as expected with a strongly anharmonic potential and in agreement with neutron inelastic scattering results. Decreasing hydration shifts the absorption to higher frequencies, possibly resulting from decreased coupling mediated by the interior water molecules. Ground-state FIR absorbances have nearly identical frequency dependence, with the mutant having less optical density than the WT. In the M state, the FIR absorbance of the WT increases whereas there is no change for D96N. These results represent the first measurement of FIR absorbance change as a function of conformational state. PMID- 12885671 TI - Ordered and disordered phases coexist in plasma membrane vesicles of RBL-2H3 mast cells. An ESR study. AB - Four chain spin labels and a spin-labeled cholestane were used to study the dynamic structure of plasma membrane vesicles (PMV) prepared from RBL-2H3 mast cells at temperatures ranging from 22 degrees C to 45 degrees C. Analysis shows that the spectra from most labels consist of two components. The abundant spectral components exhibit substantial ordering that is intermediate between that of a liquid-ordered (Lo) phase, and that of a liquid-crystalline (Lc) phase as represented by model membranes. Also, rotational diffusion rates of the spin labels are comparable to those in the Lo phase. In contrast, the ordering for the less abundant components is much lower. These results indicate that a Lo-like region or phase (the abundant component) and an Lc-like region or phase (the less abundant component) coexist in the PMV. In contrast, membranes reconstituted from extracted lipids exhibit the more ordered phase only. This suggests that membrane associated proteins are important for the coexistence of Lo-like and Lc-like regions in the plasma membrane. In addition, binding of the myristoylated protein, ARF6 to PMV, leads to a new spectral component for a headgroup lipid spin label that indicates the formation of plasma membrane defects by this low molecular weight GTPase. PMID- 12885672 TI - Recovery, visualization, and analysis of actin and tubulin polymer flow in live cells: a fluorescent speckle microscopy study. AB - Fluorescent speckle microscopy (FSM) is becoming the technique of choice for analyzing in vivo the dynamics of polymer assemblies, such as the cytoskeleton. The massive amount of data produced by this method calls for computational approaches to recover the quantities of interest; namely, the polymerization and depolymerization activities and the motions undergone by the cytoskeleton over time. Attempts toward this goal have been hampered by the limited signal-to-noise ratio of typical FSM data, by the constant appearance and disappearance of speckles due to polymer turnover, and by the presence of flow singularities characteristic of many cytoskeletal polymer assemblies. To deal with these problems, we present a particle-based method for tracking fluorescent speckles in time-lapse FSM image series, based on ideas from operational research and graph theory. Our software delivers the displacements of thousands of speckles between consecutive frames, taking into account that speckles may appear and disappear. In this article we exploit this information to recover the speckle flow field. First, the software is tested on synthetic data to validate our methods. We then apply it to mapping filamentous actin retrograde flow at the front edge of migrating newt lung epithelial cells. Our results confirm findings from previously published kymograph analyses and manual tracking of such FSM data and illustrate the power of automated tracking for generating complete and quantitative flow measurements. Third, we analyze microtubule poleward flux in mitotic metaphase spindles assembled in Xenopus egg extracts, bringing new insight into the dynamics of microtubule assemblies in this system. PMID- 12885673 TI - Low dielectric permittivity of water at the membrane interface: effect on the energy coupling mechanism in biological membranes. AB - Protonmotive force (the transmembrane difference in electrochemical potential of protons, ) drives ATP synthesis in bacteria, mitochondria, and chloroplasts. It has remained unsettled whether the entropic (chemical) component of relates to the difference in the proton activity between two bulk water phases (deltapH(B)) or between two membrane surfaces (deltapH(S)). To scrutinize whether deltapH(S) can deviate from deltapH(B), we modeled the behavior of protons at the membrane/water interface. We made use of the surprisingly low dielectric permittivity of interfacial water as determined by O. Teschke, G. Ceotto, and E. F. de Souza (O. Teschke, G. Ceotto, and E. F. de Sousa, 2001, PHYS: Rev. E. 64:011605). Electrostatic calculations revealed a potential barrier in the water phase some 0.5-1 nm away from the membrane surface. The barrier was higher for monovalent anions moving toward the surface (0.2-0.3 eV) than for monovalent cations (0.1-0.15 eV). By solving the Smoluchowski equation for protons spreading away from proton "pumps" at the surface, we found that the barrier could cause an elevation of the proton concentration at the interface. Taking typical values for the density of proton pumps and for their turnover rate, we calculated that a potential barrier of 0.12 eV yielded a steady-state pH(S) of approximately 6.0; the value of pH(S) was independent of pH in the bulk water phase under neutral and alkaline conditions. These results provide a rationale to solve the long lasting problem of the seemingly insufficient protonmotive force in mesophilic and alkaliphilic bacteria. PMID- 12885674 TI - Sucrose and glycerol effects on photosystem II. AB - Photosystem II catalyzes the oxidation of water and the reduction of plastoquinone. The active site cycles among five oxidation states, which are called the S(n) states. PSII purification procedures include the use of the cosolvents, sucrose and/or glycerol, to stabilize water splitting activity and for cryoprotection. In this study, the effects of sucrose and glycerol on PSII were investigated. Sucrose addition was observed to stimulate the steady-state rate of oxygen evolution in the range from 0 to 1.35 M. Glycerol addition was observed to stimulate oxygen evolution in the range from 0 to 30%. Both cosolvents were observed to be inhibitory at higher concentrations. Sucrose addition was shown to have no effect on the rate of Q(A)(-) oxidation or on the K(M) for exogenous acceptor. PSII was then treated to remove extrinsic proteins. In these samples, sucrose addition stimulated activity, but glycerol addition was inhibitory at concentrations higher than approximately 0.5 M. This inhibitory effect of glycerol at relatively low concentrations is attributed to glycerol binding to the active site, when extrinsic subunits are not present. Reaction induced FTIR spectra, associated with the S(1) to S(2) transition of the water oxidizing complex, exhibited significant differences throughout the 1,800-1,200 cm(-1) region, when glycerol- and sucrose-containing samples were compared. These measurements suggest a cosolvent-induced shift in the pK(A) of an aspartic or glutamic acid side chain, as well as structural changes at the active site. These structural alterations are attributed to a change in preferential hydration of the oxygen-evolving complex. PMID- 12885677 TI - What is the price of life and why doesn't it increase at the rate of inflation? PMID- 12885675 TI - Distinct potentiation of L-type currents and secretion by cAMP in rat chromaffin cells. AB - We have investigated the potentiating action of cAMP on L-currents of rat chromaffin cells and the corresponding increase of Ca(2+)-evoked secretory responses with the aim of separating the action of cAMP on Ca(2+) entry through L channels and the downstream effects of cAMP/protein kinase A (PKA) on exocytosis. In omega-toxin-treated rat chromaffin cells, exposure to the permeable cAMP analog 8-(4-chlorophenylthio)-adenosine 3',5'-monophosphate (pCPT-cAMP; 1 mM, 30 min) caused a moderate increase of Ca(2+) charge carried through L-channels (19% in 10 mM Ca(2+) at +10 mV) and a drastic potentiation of secretion ( approximately 100%), measured as membrane capacitance increments (deltaC). The apparent Ca(2+) dependency of exocytosis increased with pCPT-cAMP and was accompanied by 83% enhancement of the readily releasable pool of vesicles with no significant change of the probability of release, as evaluated with paired-pulse stimulation protocols. pCPT-cAMP effects could be mimicked by stimulation of beta(1)-adrenoreceptors and reversed by the PKA inhibitor H89, suggesting strict PKA dependence. For short pulses to +10 mV (100 ms), potentiation of exocytosis by pCPT-cAMP was proportional to the quantity of charge entering the cell and occurred independently of whether L, N, or P/Q channels were blocked, suggesting that cAMP acts as a constant amplification factor for secretion regardless of the channel type carrying Ca(2+). Analysis of statistical variations among depolarization-induced capacitance increments indicates that pCPT-cAMP acts downstream of Ca(2+) entry by almost doubling the mean size of unitary exocytic events, most likely as a consequence of an increased granule-to-granule rather than a granule-to-membrane fusion. PMID- 12885678 TI - "Inappropriate" treatment near the end of life: conflict between religious convictions and clinical judgment. AB - Not infrequently, Christian patients and families provide religious justifications for an insistence on aggressive medical care near the end of life. Four commonly invoked reasons are (1). hope for a miracle, (2). refusal to give up on the God of faith, (3). a conviction that every moment of life is a gift from God and is worth preserving at any cost, and (4). a belief that suffering can have redemptive value. For each of these 4 reasons, however, there are alternative Christian interpretations that point in the direction of limiting medical intervention under certain circumstances. When clinicians believe that an intervention is medically inappropriate or inhumane, they are not necessarily obligated to provide it simply because it is demanded on religious grounds. Instead, clinicians-preferably assisted by chaplains or clergy-should discuss alternative religious interpretations with the patient or family, and should attempt to reach a consensus on the appropriate limits to life-sustaining treatment. PMID- 12885679 TI - Neuroprotection in Parkinson disease. AB - Treatment of Parkinson disease has improved dramatically over the past quarter of a century and promising therapies are emerging. Although treatment with levodopa results in marked symptomatic improvement, mortality rates of the disease have remained relatively unchanged. Recent findings of abnormal protein folding, coupled with oxidative stress, provide scientific rationale for novel therapeutic strategies designed to slow disease progression. To be effective, these disease modifying and neuroprotective therapies must be instituted early in the course of the disease and early diagnosis therefore is critical. Consequently, primary care physicians will play an increasingly important role in early institution of such neuroprotective strategies. This review is designed to highlight some of the recent advances in our understanding of the mechanisms of neurodegeneration and to draw attention to the importance of early recognition and implementation of the new therapeutic interventions. PMID- 12885680 TI - A pilot randomized double-blind comparison of a low-molecular-weight heparin, a nonsteroidal anti-inflammatory agent, and placebo in the treatment of superficial vein thrombosis. AB - BACKGROUND: The efficacy and safety of antithrombotic treatment in patients with superficial vein thrombosis remain to be established in adequately designed trials. METHODS: In a double-blind trial, 427 patients older than 18 years, with documented acute symptomatic superficial vein thrombosis of the legs, were randomly assigned to receive subcutaneous enoxaparin sodium, 40 mg; subcutaneous enoxaparin, 1.5 mg/kg; oral tenoxicam; or placebo, once daily for 8 to 12 days. The primary efficacy outcome was deep venous thromboembolism between days 1 and 12, defined as deep vein thrombosis detected by ultrasonography between days 8 and 12 or earlier if clinically indicated, or documented symptomatic pulmonary embolism. For the secondary efficacy outcomes, superficial vein thrombosis recurrence or extension was also considered. RESULTS: The incidence of deep venous thromboembolism by day 12 was 3.6% (4 of 111 patients) in the placebo group, 0.9% (1 of 109 patients) in the 40-mg enoxaparin group (P =.37 vs placebo), 1.0% (1 of 102 patients) in the 1.5-mg/kg enoxaparin group (P =.37 vs placebo), and 2.1% (2 of 94 patients) in the tenoxicam group (P =.69 vs placebo). The incidence of deep and superficial venous thromboembolism by day 12 was significantly reduced in all active treatment groups, from 30.6% (34 of 111 patients) in the placebo group to 8.3% (9 of 109 patients), 6.9% (7 of 102 patients), and 14.9% (14 of 94 patients) in the 40-mg enoxaparin (P<.001), 1.5 mg/kg enoxaparin (P<.001), and tenoxicam (P<.01) groups, respectively. No death or major hemorrhage occurred during the study. CONCLUSION: Treatment with a low molecular-weight heparin or with an oral nonsteroidal anti-inflammatory agent should be evaluated further in the prevention of thromboembolic complications in patients with superficial vein thrombosis. PMID- 12885681 TI - Impact of oseltamivir treatment on influenza-related lower respiratory tract complications and hospitalizations. AB - BACKGROUND: Influenza causes lower respiratory tract complications (LRTCs), particularly bronchitis and pneumonia, in both otherwise healthy adults and those with underlying conditions. The aim of this study was to assess the effect of oseltamivir treatment on the incidence of LRTCs leading to antibiotic treatment and hospitalizations following influenza illness. METHODS: We analyzed prospectively collected data on LRTCs and antibiotic use from 3564 subjects (age range, 13-97 years) with influenzalike illness enrolled in 10 placebo-controlled, double-blind trials of oseltamivir treatment. RESULTS: In adults and adolescents with a proven influenza illness, oseltamivir treatment reduced overall antibiotic use for any reason by 26.7% (14.0% vs 19.1% with placebo; P<.001) and the incidence of influenza-related LRTCs resulting in antibiotic therapy by 55% (4.6% vs 10.3% with placebo; P<.001). In those subjects considered at increased risk of complications, 74 (18.5%) of 401 placebo recipients developed an LRTC leading to antibiotic use compared with 45 (12.2%) of 368 oseltamivir recipients (34.0% reduction; P =.02). Hospitalization for any cause occurred in 18 (1.7%) of 1063 placebo recipients compared with 9 (0.7%) of 1350 oseltamivir-treated patients (59% reduction; P =.02). In contrast, among subjects with an influenzalike illness but without a confirmed influenza infection, the incidence of LRTCs (6.7% vs 5.3%), overall antibiotic use (19.7% vs 19.3%), or hospitalizations (1.7% vs 1.9%) was similar between placebo and oseltamivir recipients, respectively. CONCLUSION: Oseltamivir treatment of influenza illness reduces LRTCs, antibiotic use, and hospitalization in both healthy and "at-risk" adults. PMID- 12885682 TI - Direct observation of requests for clinical services in office practice: what do patients want and do they get it? AB - BACKGROUND: Requests can influence the conduct and content of the medical visit. However, little is known about the nature, frequency, and impact of such requests. We performed this study to ascertain the prevalence, antecedents, and consequences of patients' requests for clinical services in ambulatory practice. METHODS: This observational study combined patient and physician surveys with audiotaping of 559 visits to 45 physicians in 2 health care systems between January and November 1999. All patients had a new problem or significant health concern. Main outcome measures included prevalence of 8 categories of requests for physician action; odds of patients' requesting tests, referrals, or new prescriptions; odds of physicians' ordering diagnostic tests, making specialty referrals, or writing new prescriptions; patient satisfaction; and physicians' perceptions of the visit. RESULTS: The 559 patients made 545 audiocoded requests for physician action; 23% requested at least 1 diagnostic test, specialty referral, or new prescription medication. Requests for diagnostic tests were more common among new patients (P<.001). Requests for any clinical service were more common among patients experiencing greater health-related distress (P<.05) and less common among patients of cardiologists (P<.001). After adjusting for predisposing, need, and contextual factors, referral requests were associated with higher odds of receiving specialty referrals (adjusted odds ratio [AOR], 4.1; 95% confidence interval [CI], 1.6-10.7) and prescription requests were associated with higher odds of receiving new prescription medications (AOR, 2.8; 95% CI, 1.2-6.3). Physicians reported that visits during which patients requested diagnostic tests were more demanding than visits in which no such requests were made (P =.02). CONCLUSIONS: Though more common in primary care than in cardiology, patients' requests for clinical services are both pervasive and influential. The results support placing greater emphasis on understanding and addressing the patient's role in determining health care utilization. PMID- 12885683 TI - Cardiorespiratory fitness and the risk for stroke in men. AB - BACKGROUND: Low cardiorespiratory fitness is considered to be a major public health problem. We examined the relationship of cardiorespiratory fitness, as indicated by maximum oxygen consumption VO(2)max with subsequent incidence of stroke. We also compared VO(2)max with conventional risk factors as a predictor for future strokes. METHODS: Population-based cohort study with an average follow up of 11 years from Kuopio and surrounding communities of eastern Finland. Of 2011 men with no stroke or pulmonary disease at baseline who participated in the study, 110 strokes occurred, of which 87 were ischemic. The VO(2)max was measured directly during exercise testing at baseline. RESULTS: The relative risk for any stroke in unfit men VO(2)max, <25.2 mL/kg per minute) was 3.2 (95% confidence interval [CI], 1.71-6.12; P<.001; P<.001 for the trend across the quartiles); and for ischemic stroke, 3.50 (95% CI, 1.66-7.41; P =.001; P<.001 for trend across the quartiles), compared with fit men VO(2)max, >35.3 mL/kg per minute), after adjusting for age and examination year. The associations remained statistically significant after further adjustment for smoking, alcohol consumption, socioeconomic status, energy expenditure of physical activity, prevalent coronary heart disease, diabetes, systolic blood pressure, and serum low-density lipoprotein cholesterol level for any strokes or ischemic strokes. Low cardiorespiratory fitness was comparable with systolic blood pressure, obesity, alcohol consumption, smoking, and serum low-density lipoprotein cholesterol level as a risk factor for stroke. CONCLUSIONS: Our findings show that low cardiorespiratory fitness was associated with an increased risk for any stroke and ischemic stroke. The VO(2)max was one of the strongest predictors of stroke, comparable with other modifiable risk factors. PMID- 12885684 TI - Venous thromboembolic disease: comparison of the diagnostic process in men and women. AB - BACKGROUND: There has been concern that women may be limited to fewer major diagnostic tests than men. Whether this applies to patients with pulmonary embolism (PE) or deep venous thrombosis (DVT) has not been determined. OBJECTIVE: To assess whether there is a sex disparity in the application of diagnostic tests for PE or DVT, in reaching a diagnosis, or in using medical facilities. DESIGN: A study of cross-sectional samples of hospitalizations from 21 separate years using data from the National Hospital Discharge Survey. SETTING: Noninstitutional hospitals in the 50 states and the District of Columbia from 1979 through 1999. PATIENTS: The National Hospital Discharge Survey abstracts demographic and medical information from the medical records of inpatients. For 1979 through 1999, the number of patients sampled ranged from 181000 to 307000.Measurements The number of sampled patients with DVT and with PE and the number of diagnostic tests performed were determined from the International Classification of Diseases, Ninth Revision, Clinical Modification codes at discharge. A multistage estimation procedure gave an estimation of values for the entire United States. RESULTS: Age-adjusted rates of the diagnosis of PE per 100000 population and of DVT per 100000 population were not lower in women. Rates of the use of ventilation-perfusion lung scans, venous ultrasonography of the lower extremities, and contrast venography were not lower in women. Durations of hospitalization for PE or DVT were comparable in men and women. CONCLUSION: Data from the National Hospital Discharge Survey do not support a sex bias in the diagnosis of PE or DVT, the use of diagnostic tests, or the duration of hospitalization for PE or DVT. PMID- 12885685 TI - Initial and maintenance naltrexone treatment for alcohol dependence using primary care vs specialty care: a nested sequence of 3 randomized trials. AB - BACKGROUND: Naltrexone may improve success in primary care treatment of alcohol dependence (AD). This study tests naltrexone and primary care management (PCM) vs naltrexone and cognitive behavior therapy (CBT) and tests naltrexone maintenance among patients who respond to an initial course of naltrexone combined with PCM vs CBT. METHODS: A nested sequence of 3 randomized trials was conducted. In study 1, 197 subjects with AD participated in a 10-week comparison of PCM and naltrexone (50 mg/d) vs CBT and naltrexone (50 mg/d). In study 2, 53 PCM responders from study 1 continued in a 24-week placebo-controlled study of maintenance naltrexone. In study 3, 60 CBT responders from study 1 continued in a 24-week placebo-controlled study of maintenance naltrexone and CBT. RESULTS: Study 1: No difference in the response to treatment; 84.1% (74/88) of the PCM patients and 86.5% (77/89) of the CBT patients avoided persistent heavy drinking. Percentage of days abstinent (PDA) declined over time for PCM vs CBT (P =.03). Study 2: Higher response maintenance for PCM and naltrexone (21/26, 80.8%) vs PCM and placebo (14/27, 51.9%; P =.03) and PDA declined more for the placebo group (P =.02). Study 3: The differences between naltrexone vs placebo on maintenance of response (25/30, 83.3% vs 21/30, 70.0%) or PDA did not reach statistical significance. CONCLUSIONS: Naltrexone yielded comparable results during the initial 10 weeks of treatment when combined with PCM or CBT. Maintenance of improvement was enhanced by continued naltrexone treatment in the PCM but not in the CBT arm. PMID- 12885686 TI - Predictors of warfarin use among Ohio medicaid patients with new-onset nonvalvular atrial fibrillation. AB - BACKGROUND: Despite demonstrated efficacy in stroke prevention, warfarin is underused in patients with atrial fibrillation (AF). Reasons for warfarin nonuse are unclear. METHODS: We conducted a retrospective cohort analysis using Ohio Medicaid administrative billing data to ascertain determinants of warfarin use for patients with new-onset nonvalvular AF. The database included data from all institutions, providers, and pharmacies providing services to Ohio Medicaid enrollees. Subjects included all 11699 continuously enrolled fee-for-service recipients of Ohio Medicaid with a new diagnosis of nonvalvular AF between January 1, 1998, and December 31, 2000. We determined incipient warfarin use and presence of risk factors for stroke and hemorrhage by searching claims records for corresponding International Classification of Diseases, Ninth Revision, Clinical Modification codes and National Drug Codes. Univariate and multivariable analyses were performed to examine the association of risk factors with warfarin use. RESULTS: Only 9.7% of all patients and 11.9% of those without apparent contraindications filled prescriptions for warfarin from 7 days preceding to 30 days after the development of AF. Hypertension and congestive heart failure independently predicted increased warfarin use. Older age (>or=85 years), younger age (<55 years), prior intracranial hemorrhage, prior gastrointestinal hemorrhage, predisposition to falls, alcohol or other drug abuse, renal impairment, and conditions perceived as barriers to compliance predicted decreased warfarin use. CONCLUSIONS: Few in this cohort of Ohio Medicaid patients with incident AF filled prescriptions for warfarin within 30 days of the diagnosis. Several factors, including alcohol or other drug abuse or dependence, psychiatric disease, homelessness or inadequate housing, and lack of a caregiver, were highly prevalent and seemed to bias against warfarin prescribing. PMID- 12885687 TI - Pulmonary embolism mortality in the United States, 1979-1998: an analysis using multiple-cause mortality data. AB - BACKGROUND: Pulmonary thromboembolism (PTE) is a common clinical problem that is associated with substantial morbidity and mortality. Estimates of PTE mortality and predictions of PTE trends have varied widely. These estimates play a role in the planning of national health strategies. The analysis of pulmonary embolism mortality trends and comorbidities may elucidate how well we treat and prevent the disease as well as identify additional risk factors. METHODS: We analyzed PTE (International Classification of Diseases, Ninth Revision code 415.1) as reported on death certificates in the Multiple-Cause Mortality Files compiled by the National Center for Health Statistics from 1979 to 1998. RESULTS: Of all the 42932973 decedents, 572773 (1.3%) had PTE listed on their death certificates and 194389 of these (33.9%) had PTE as the underlying cause of death. The age adjusted rate of deaths with PTE decreased from 191 per million in 1979 to 94 per million in 1998 overall, decreasing 56% for men and 46% for women. During the study period, the age-adjusted mortality rates for blacks were consistently 50% higher than those for whites, and those for whites were 50% higher than those for people of other races (Asian, American Indian, etc). Within racial strata, mortality rates were consistently 20% to 30% higher among men than among women. Conditions that were of higher likelihood in persons who died with PTE included thrombophlebitis, fractures, trauma, postoperative complications, certain cancers, and the inflammatory bowel diseases. CONCLUSIONS: Mortality with PTE in the United States has decreased during the 20-year period. The mortality rates between men and women and between racial groups vary substantially. These findings may be useful in better directing preventive therapy efforts. PMID- 12885688 TI - Azithromycin monotherapy for patients hospitalized with community-acquired pneumonia: a 31/2-year experience from a veterans affairs hospital. AB - BACKGROUND: Current American Thoracic Society (ATS) community-acquired pneumonia treatment guidelines recommend azithromycin monotherapy for a limited subset of hospitalized patients. We evaluated the effectiveness of azithromycin monotherapy in a more generalized population of patients hospitalized with mild-to-moderate community-acquired pneumonia. METHODS: We reviewed medical records from a Veterans Affairs facility for patients admitted with community-acquired pneumonia between December 1, 1997, and June 30, 2001, comparing those receiving azithromycin monotherapy, other ATS-recommended antibiotics, and non-ATS recommended antibiotics. We excluded patients with immunosuppression, metastatic cancer, or hospital-acquired pneumonia. Outcome measures included times to stability, meeting criteria for change to oral therapy, and eligibility for discharge; length of stay; intensive care unit transfer; and mortality. Outcomes were adjusted for pneumonia severity, skilled nursing facility status, and processes of care. RESULTS: A total of 442 patients were eligible for the study (221 in the azithromycin monotherapy group, 129 in the ATS group, and 92 in the non-ATS group). Times to clinical stability and to fulfilling early switch criteria were not statistically significantly different among the 3 groups. Mean time to fulfilling early discharge criteria was 2.48 days for patients receiving azithromycin monotherapy vs 2.84 days for those receiving ATS antibiotics (P =.008) and 2.58 days for those receiving non-ATS antibiotics (P =.64). Overall mean length of stay was shorter in the azithromycin monotherapy group (4.35 days) vs the ATS (5.73 days) (P =.002) and non-ATS (6.21 days) (P<.001) groups. Mortality, intensive care unit transfer, and readmission rates were similar across the groups. CONCLUSION: Azithromycin monotherapy is equally as efficacious as other ATS-recommended regimens for treating hospitalized patients with mild-to moderate community-acquired pneumonia. PMID- 12885689 TI - The impact of health information on the internet on the physician-patient relationship: patient perceptions. AB - BACKGROUND: Use of the Internet for health information continues to grow rapidly, but its impact on health care is unclear. Concerns include whether patients' access to large volumes of information will improve their health; whether the variable quality of the information will have a deleterious effect; the effect on health disparities; and whether the physician-patient relationship will be improved as patients become more equal partners, or be damaged if physicians have difficulty adjusting to a new role. METHODS: Telephone survey of nationally representative sample of the American public, with oversample of people in poor health. RESULTS: Of the 3209 respondents, 31% had looked for health information on the Internet in the past 12 months, 16% had found health information relevant to themselves and 8% had taken information from the Internet to their physician. Looking for information on the Internet showed a strong digital divide; however, once information had been looked for, socioeconomic factors did not predict other outcomes. Most (71%) people who took information to the physician wanted the physician's opinion, rather than a specific intervention. The effect of taking information to the physician on the physician-patient relationship was likely to be positive as long as the physician had adequate communication skills, and did not appear challenged by the patient bringing in information. CONCLUSIONS: For health information on the Internet to achieve its potential as a force for equity and patient well-being, actions are required to overcome the digital divide; assist the public in developing searching and appraisal skills; and ensure physicians have adequate communication skills. PMID- 12885690 TI - Sex differences in risk for coronary heart disease mortality associated with diabetes and established coronary heart disease. AB - BACKGROUND: The sex-specific independent effect of diabetes mellitus and established coronary heart disease (CHD) on subsequent CHD mortality is not known. METHODS: This is an analysis of pooled data (n = 5243) from the Framingham Heart Study and the Framingham Offspring Study with follow-up of 20 years. At baseline (1971-1975), 134 men and 95 women had diabetes, while 222 men and 129 women had CHD. Risk for CHD death was analyzed by proportional hazards models, adjusting for age, hypertension, serum cholesterol levels, smoking, and body mass index. The comparative effect of established CHD vs diabetes on the risk of CHD mortality was tested by testing the difference in log hazards. RESULTS: The adjusted hazard ratios (HRs) with 95% confidence intervals (CIs) for death from CHD were 2.1 (95% CI, 1.3-3.3) in men with diabetes only, and 4.2 (95% CI, 3.2 5.6) in men with CHD only compared with men without diabetes or CHD. The HR for CHD death was 3.8 (95% CI, 2.2-6.6) in women with diabetes, and 1.9 (95% CI, 1.1 3.4) in women with CHD. The difference between the CHD and the diabetes log hazards was +0.73 (95% CI, 0.72-0.75) in men and -0.65 (95% CI, -0.68 to -0.63) in women. CONCLUSIONS: In men, established CHD signifies a higher risk for CHD mortality than diabetes. This is reversed in women, with diabetes being associated with greater risk for CHD mortality. Current treatment recommendations for women with diabetes may need to be more aggressive to match CHD mortality risk. PMID- 12885691 TI - Should diabetic patients treated long-term with sulfonylureas be switched to nateglinide? PMID- 12885692 TI - Professionalism: a barrier to provision of quality health care? PMID- 12885693 TI - Prevalence of viral myocarditis. PMID- 12885694 TI - Questioning race-based hypertension management. PMID- 12885695 TI - Who actually has the "low health literacy"? PMID- 12885696 TI - National Health and Nutrition Examination Survey: where nutrition meets medicine for the benefit of health. PMID- 12885697 TI - A plea for prevention. PMID- 12885698 TI - Nutrition and genetics: an expanding frontier. AB - The age of molecular biology began in 1953 with the discovery of the structure of DNA. By 1961 the genetic code for the translation of the sequence of bases in DNA to amino acids in proteins was underway, and a model for the genetic regulation of protein synthesis was proposed. My interest in the genetic regulation of nutrient metabolism began in that year during my sabbatical leave in the laboratory of Sir Hans Krebs at Oxford University. In the present article, I describe 2 episodes in my career during which I used genetic concepts to explain a nutritional phenomenon; the first episode occurred before doing the experimental work, and the second occurred after the experimental work was completed. My first brainstorm, which occurred in 1961, was to investigate the hypothesis that all of the fat-soluble vitamins act by the regulation of a cluster of genes. Unfortunately, I selected vitamin K as my model and discovered that it is the only fat-soluble vitamin that does not work in full or in part by the regulation of a set of genes. In 1967 I undertook a second problem, which was to determine the mode of action of polyunsaturated fatty acids in lowering plasma lipid concentrations in humans. We discovered that linoleic acid reduced the storage and enhanced the oxidation of fatty acids. The genetic interpretation of this study has come only recently: polyunsaturated fats have been shown to down regulate enzymes that accomplish storage of fatty acids and to up-regulate genes that enhance fatty acid oxidation. PMID- 12885699 TI - Leisure-time activity is an important determinant of long-term weight maintenance after weight loss in the Sibutramine Trial on Obesity Reduction and Maintenance (STORM trial). AB - BACKGROUND: The success rate of long-term maintenance of weight loss in obese patients is usually low. To improve the success rate, determinants of long-term weight maintenance must be identified. OBJECTIVE: The objective of the study was to identify determinants of long-term success in weight maintenance in obese subjects who completed the Sibutramine Trial on Obesity Reduction and Maintenance (n = 261), a multicenter European study of weight loss and weight maintenance in obesity that combines sibutramine treatment with dietary restriction and advice on exercise and behavior. DESIGN: We studied weight maintenance over 18 mo in subjects who had completed a 6-mo weight-loss phase. Factors included in the analysis were initial body weight, the percentage of initial body weight lost, dietary intake, various components of physical activity (measured with the Baecke questionnaire), the type of treatment (sibutramine or placebo), age, and sex. RESULTS: Multiple regression analysis identified treatment group (sibutramine or placebo), the percentage of the initial body weight that was lost during the 6-mo weight-loss phase, and the leisure-time physical activity index as significant determinants of weight maintenance. Together, these 3 factors explained 20% of the variation in weight maintenance (P < 0.001). Dietary factors, age, and sex were not significant predictors of weight-maintenance success in this study. CONCLUSIONS: Weight-maintenance success after weight loss is positively influenced by sibutramine treatment during weight maintenance, by a greater initial weight loss, and by a higher leisure-time physical activity index, which reflects higher levels of activities such as walking and cycling and lower levels of television viewing. PMID- 12885700 TI - Learning to overeat: maternal use of restrictive feeding practices promotes girls' eating in the absence of hunger. AB - BACKGROUND: Experimental findings causally link restrictive child-feeding practices to overeating in children. However, longitudinal data are needed to determine the extent to which restrictive feeding practices promote overeating. OBJECTIVES: Our objectives were to determine whether restrictive feeding practices foster girls' eating in the absence of hunger (EAH) and whether girls' weight status moderates the effects of restrictive feeding practices. DESIGN: Longitudinal data were used to create a study design featuring 2 maternal restriction factors (low and high), 2 weight-status factors (nonoverweight and overweight), and 3 time factors (ages 5, 7, and 9 y). RESULTS: Mean EAH increased significantly (P < 0.0001) from 5 to 9 y of age. Higher levels of restriction at 5 y of age predicted higher EAH at 7 y of age (P < 0.001) and at 9 y of age (P < 0.01). Girls who were already overweight at 5 y of age and who received higher levels of restriction had the highest EAH scores at 9 y of age (P < 0.05) and the greatest increases in EAH from 5 to 9 y of age (P < 0.01). CONCLUSIONS: The developmental increase in EAH from 5 to 9 y of age may be especially problematic in obesigenic environments. These longitudinal data provide evidence that maternal restriction can promote overeating. Girls who are already overweight at 5 y of age may be genetically predisposed to be especially responsive to environmental cues. These findings are not expected to be generalized to boys or to other racial and ethnic groups. PMID- 12885701 TI - Cholesterol-lowering effect of beta-glucan from oat bran in mildly hypercholesterolemic subjects may decrease when beta-glucan is incorporated into bread and cookies. AB - BACKGROUND: Findings about the effects of beta-glucan on serum lipoproteins are conflicting. OBJECTIVE: The study investigated the effects of beta-glucan from oat bran in bread and cookies (study 1) and in orange juice (study 2) on serum lipoproteins in mildly hypercholesterolemic subjects. DESIGN: In study 1, 48 subjects (21 men, 27 women) received for 3 wk control bread and cookies rich in wheat fiber. For the next 4 wk, by random assignment, 23 subjects continued to consume the control products, and 25 received bread and cookies rich in beta glucan. Mean daily intake of beta-glucan was 5.9 g. Total dietary fiber intake did not differ significantly between the groups. In study 2, the same sources of control fiber and beta-glucan (5 g/d) as in study 1 were provided. For 2 wk, 25 of the original 48 subjects (10 men, 15 women) were randomly assigned to consume orange juice containing either wheat fiber (n = 13) or beta-glucan from oat bran (n = 12). After a washout period of 1 wk, dietary regimens were crossed over. RESULTS: In study 1, the change in LDL cholesterol did not differ significantly ( 0.12 mmol/L; P = 0.173) between the 2 groups. In study 2, the drink rich in beta glucan decreased LDL cholesterol by 0.26 +/- 0.07 mmol/L (6.7 +/- 1.8%; P = 0.001) and the ratio of total to HDL cholesterol by 0.26 +/- 0.11 (5.4 +/- 2.1%; P = 0.029) compared with the other drink. HDL-cholesterol and triacylglycerol concentrations did not change significantly. CONCLUSIONS: The food matrix or the food processing, or both, could have adverse effects on the hypocholesterolemic properties of oat beta-glucan. PMID- 12885702 TI - Percentage body fat ranges associated with metabolic syndrome risk: results based on the third National Health and Nutrition Examination Survey (1988-1994). AB - BACKGROUND: Increasing attention has focused on the association between body fatness and related metabolic risk factors. The quantitative link between percentage body fat (%BF) and the risk of metabolic syndrome is unknown. OBJECTIVES: The objectives were to determine the risk [odds ratios (ORs)] of metabolic syndrome based on %BF in black and white men and women in the United States and to provide corresponding ranges of %BF associated with a risk equivalent to body mass index (BMI; in kg/m(2)). DESIGN: The subjects were participants in the third National Health and Nutrition Examination Survey and were divided into those with and without the metabolic syndrome. OR equations were derived from logistic regression models for %BF and BMI, with the 25th percentile in the study population as the reference. Ranges were developed by associating %BF with the equivalent risk of metabolic syndrome based on established BMI cutoffs. RESULTS: Four sets (men, women, black, and white) of OR curves were generated for %BF and for BMI by using data from 8259 adults. The ORs for metabolic syndrome were lower in blacks than in whites at any given %BF or BMI. The developed cutoffs for %BF differed between men and women but showed only small race and age effects. A simplified set of sex-specific %BF ranges for the risk of metabolic syndrome were developed. CONCLUSIONS: The risk of metabolic syndrome can be established from measured %BF by using either the developed OR curves or %BF thresholds at traditional BMI cutoffs. This information should prove useful in both clinical and research settings. PMID- 12885703 TI - Interchangeability of pencil-beam and fan-beam dual-energy X-ray absorptiometry measurements in piglets and infants. AB - BACKGROUND: Compared with the older pencil-beam (PB) dual-energy X-ray absorptiometry (DXA), the newer fan-beam (FB) DXA has the advantage of faster scan acquisition and greater accuracy of body-composition measurement in small subjects. However, no data exist on the relation between the measurements obtained with these techniques. OBJECTIVE: The objective of the study was to investigate whether PB and FB DXA measurements in small subjects are interchangeable. DESIGN: PB and FB DXA scans were performed on 26 piglets and 54 infants to examine the relation between the measurements obtained by using the 2 techniques. RESULTS: The correlation between all PB and FB DXA measurements of variables (total weight, bone area, bone mineral content, bone mineral density, and lean and fat masses) approached 1.0, but there were significant differences in absolute values. The extent of the differences varied according to the variable, with the lowest value for total weight (mean difference: approximately 1% for both piglets and infants) and the highest value for bone mineral content (mean difference: 35.3% and 36.7% for piglets and infants, respectively). PB and FB DXA measurements were strongly predictive of each other after adjustment (r(2) = 0.927-1.000 for the piglet data and 0.939-0.999 for the infant data). CONCLUSION: In small subjects, DXA measurements from PB and FB techniques were strongly predictive of each other, although their absolute values differed. Thus, group comparison of PB and FB DXA data is possible after adjustment of the data from either technique. It is advisable to generate normative data for each technique and to use the same technique throughout longitudinal studies. PMID- 12885704 TI - Assessment of resting energy expenditure in mechanically ventilated patients. AB - BACKGROUND: Usual equations for predicting resting energy expenditure (REE) are not appropriate for critically ill patients, and indirect calorimetry criteria render its routine use difficult. OBJECTIVE: Variables that might influence the REE of mechanically ventilated patients were evaluated to establish a predictive relation between these variables and REE. DESIGN: The REE of 70 metabolically stable, mechanically ventilated patients was prospectively measured by indirect calorimetry and calculated with the use of standard predictive models (Harris and Benedict's equations corrected for hypermetabolism factors). Patient data that might influence REE were assessed, and multivariate analysis was conducted to determine the relations between measured REE and these data. Measured and calculated REE were compared by using the Bland-Altman method. RESULTS: Multivariate analysis retained 4 independent variables defining REE: body weight (r(2) = 0.14, P < 0.0001), height (r(2) = 0.11, P = 0.0002), minute ventilation (r(2) = 0.04, P = 0.01), and body temperature (r(2) = 0.07, P = 0.002): REE (kcal/d) = 8 x body weight + 14 x height + 32 x minute ventilation + 94 x body temperature - 4834. REE calculated with this equation was well correlated with measured REE (r(2) = 0.61, P < 0.0001). Bland-Altman plots showed a mean bias approaching zero, and the limits of agreement between measured and predicted REE were clinically acceptable. CONCLUSION: Our results suggest that REE estimated on the basis of body weight, height, minute ventilation, and body temperature is clinically more relevant than are the usual predictive equations for metabolically stable, mechanically ventilated patients. PMID- 12885705 TI - Essential amino acids are primarily responsible for the amino acid stimulation of muscle protein anabolism in healthy elderly adults. AB - BACKGROUND: Nutritional supplementation may be used to treat muscle loss with aging (sarcopenia). However, if physical activity does not increase, the elderly tend to compensate for the increased energy delivered by the supplements with reduced food intake, which results in a calorie substitution rather than supplementation. Thus, an effective supplement should stimulate muscle anabolism more efficiently than food or common protein supplements. We have shown that balanced amino acids stimulate muscle protein anabolism in the elderly, but it is unknown whether all amino acids are necessary to achieve this effect. OBJECTIVE: We assessed whether nonessential amino acids are required in a nutritional supplement to stimulate muscle protein anabolism in the elderly. DESIGN: We compared the response of muscle protein metabolism to either 18 g essential amino acids (EAA group: n = 6, age 69 +/- 2 y; +/- SD) or 40 g balanced amino acids (18 g essential amino acids + 22 g nonessential amino acids, BAA group; n = 8, age 71 +/- 2 y) given orally in small boluses every 10 min for 3 h to healthy elderly volunteers. Muscle protein metabolism was measured in the basal state and during amino acid administration via L-[ring-(2)H(5)]phenylalanine infusion, femoral arterial and venous catheterization, and muscle biopsies. RESULTS: Phenylalanine net balance (in nmol x min(-1). 100 mL leg volume(-1)) increased from the basal state (P < 0.01), with no differences between groups (BAA: from -16 +/- 5 to 16 +/- 4; EAA: from -18 +/- 5 to 14 +/- 13) because of an increase (P < 0.01) in muscle protein synthesis and no change in breakdown. CONCLUSION: Essential amino acids are primarily responsible for the amino acid-induced stimulation of muscle protein anabolism in the elderly. PMID- 12885706 TI - Short-term (intestinal) and long-term (postintestinal) conversion of beta carotene to retinol in adults as assessed by a stable-isotope reference method. AB - BACKGROUND: Quantitative information on the conversion of beta-carotene to vitamin A in humans is limited. OBJECTIVE: We determined the short- and long-term conversion of labeled beta-carotene to vitamin A by using a stable-isotope reference method. DESIGN: [(2)H(8)]beta-Carotene (11,011 nmol, or 6 mg) in oil was given with a liquid diet (25% of energy from fat) to 22 adult volunteers (10 men, 12 women). Three days after the [(2)H(8)]beta-carotene dose, the volunteers each took a dose of [(2)H(8)]retinyl acetate (8915 nmol, or 3 mg) in oil with the same liquid diet. Blood samples were collected over 56 d. RESULTS: The 53-d area under the serum [(2)H(4)]retinol response curve (from the [(2)H(8)]beta-carotene dose) was 569 +/- 385 nmol. d, and the 53-d area under the serum [(2)H(8)]retinol response curve (from the [(2)H(8)]retinyl acetate dose) was 1798 +/- 1139 nmol. d. With the use of [(2)H(8)]retinyl acetate as the vitamin A reference, the [(2)H(4)]retinol formed from [(2)H(8)]beta-carotene (11,011 nmol) was calculated to be equivalent to 3413.9 +/- 2298.4 nmol retinol. The conversion factor of beta carotene to retinol varied from 2.4 to 20.2, and the average conversion factor was 9.1 to 1 by wt or 4.8 to 1 by mol. This conversion factor was positively correlated with body mass index (r = 0.57, P = 0.006). The postabsorption conversion of beta-carotene was estimated as 7.8%, 13.6%, 16.4%, and 19.0% of the total converted retinol at 6, 14, 21, or 53 d after the [(2)H(8)]beta-carotene dose, respectively. CONCLUSION: The quantitative determination of the conversion of beta-carotene to vitamin A in humans can be accomplished by using a stable isotope reference method. This approach provides in vivo metabolic information after a physiologic dose of beta-carotene. PMID- 12885707 TI - Ascorbic acid from lime juice does not improve the iron status of iron-deficient women in rural Mexico. AB - BACKGROUND: Although ascorbic acid (AA) increases dietary iron bioavailability, there has been no food-based community trial of its efficacy in improving iron status. OBJECTIVE: The objective was to assess the efficacy of 25 mg AA as agua de limon (limeade), consumed with each of 2 daily meals, in improving the iron status of iron-deficient women. DESIGN: Two rural Mexican populations were randomly assigned to an AA or a placebo group, each with 18 iron-deficient women. The AA group was given 500 mL limeade containing 25 mg AA twice a day, 6 d/wk, for 8 mo. The placebo group was given a lime-flavored beverage free of AA or citric acid. Beverages were consumed within 30 min of 2 main daily meals. Data were collected on morbidity (3 times/wk), dietary intake (on 6 d), socioeconomic status, parasites (twice), medical history, and response to treatment. Blood samples at 0, 2, 4, 6, and 8 mo were analyzed for hemoglobin, plasma AA, plasma ferritin, transferrin receptors, and C-reactive protein. RESULTS: AA intake was significantly (P < 0.0001) higher in the AA group, but nonheme iron, heme iron, and phytic acid intakes did not differ significantly. Plasma AA was significantly (P < 0.01) higher in the AA group at 2, 4, 6, and 8 mo. There were no final differences between groups in hemoglobin, plasma ferritin, or transferrin receptor concentrations or in the ratio of transferrin receptors to plasma ferritin after control for initial concentrations. CONCLUSION: Increasing dietary AA by 25 mg at each of 2 meals/d did not improve iron status in iron-deficient women consuming diets high in phytate and nonheme iron. PMID- 12885708 TI - Iron absorption from fish sauce and soy sauce fortified with sodium iron EDTA. AB - BACKGROUND: Fish sauce and soy sauce have been suggested as food vehicles for iron fortification in Asia. NaFeEDTA is a potentially useful fortificant because it can be added to these condiments without causing precipitation during storage. OBJECTIVES: The objectives were to evaluate iron absorption from NaFeEDTA fortified fish sauce and soy sauce against a reference fortificant (FeSO(4)), to compare iron absorption from NaFeEDTA-fortified fish sauce and soy sauce, and to evaluate the influence of fish sauce and soy sauce per se on iron absorption. DESIGN: Five separate iron-absorption studies were made in adult women (10 women per study). Iron absorption was measured on the basis of erythrocyte incorporation of (57)Fe or (58)Fe 14 d after the intake of labeled meals of rice or rice and vegetables. Fish sauce or soy sauce (10 g) fortified with 5 mg Fe as NaFeEDTA or FeSO(4) was fed with selected meals. The results are presented as geometric means. RESULTS: Iron absorption from NaFeEDTA- and FeSO(4)-fortified fish sauce (3.3% and 3.1%, respectively) and soy sauce (6.1% and 5.6%, respectively) was not significantly different. No significant difference was observed when NaFeEDTA-fortified fish sauce and soy sauce were compared directly (6.7% and 7.9%, respectively). Soy sauce inhibited iron absorption from rice based meals (8.5% without and 6.0% with soy sauce; P < 0.02), whereas fish sauce did not affect iron absorption significantly. CONCLUSION: The relatively high iron absorption from NaFeEDTA-fortified fish sauce and soy sauce and the acceptable organoleptic properties of NaFeEDTA indicate the potential usefulness of this iron fortificant in fish sauce and soy sauce fortification programs. PMID- 12885709 TI - Absorption of zinc from wheat products fortified with iron and either zinc sulfate or zinc oxide. AB - BACKGROUND: Several chemical forms of zinc have been proposed for food fortification, but information is needed on their absorption from common cereals having varied phytate content. OBJECTIVE: The goal was to measure zinc absorption from wheat products fortified with iron sulfate and either zinc sulfate or zinc oxide. DESIGN: Adult volunteers received either low-phytate bread (n = 11) or higher-phytate porridge (n = 11) once weekly on 2 or 3 occasions. The foods were fortified with 1 of the 2 zinc salts (60 mg elemental Zn/kg wheat flour) during week 1 and with the other during week 2, in random order. (65)Zn in the same chemical form as the fortificant was incorporated in each food to assess zinc absorption with the use of whole-body counting. The porridge group received an additional test meal fortified with zinc oxide during week 3, but the (65)Zn tracer was given as an oral solution of (65)ZnCl(2). RESULTS: Zinc absorption from bread (13.8%; 95% CI: 11.8%, 16.2%) was significantly (P < 0.001) greater than from porridge (6.4%; 5.5%, 7.6%), presumably because of the greater phytate content of the porridge. With control for food type, there were no significant differences in zinc absorption from meals fortified with zinc sulfate or zinc oxide (P = 0.24). When the porridge was fortified with zinc oxide and labeled with (65)ZnCl(2), absorption of the tracer (8.9%; 7.1%, 11.0%) was significantly (P = 0.007) greater than when (65)ZnO was incorporated in the porridge (5.6%; 4.5%, 6.9%). CONCLUSIONS: Either zinc oxide or zinc sulfate can be used to fortify wheat products consumed by presumably healthy persons. Isotopic tracers used to assess the absorption of mineral fortificants should have the same chemical form as the fortificant. PMID- 12885710 TI - Regular consumption of NaFeEDTA-fortified fish sauce improves iron status and reduces the prevalence of anemia in anemic Vietnamese women. AB - BACKGROUND: Fish sauce is consumed daily by a large proportion of the Vietnamese population and could therefore be a potentially useful food vehicle for iron fortification programs. OBJECTIVE: We evaluated the efficacy of iron-fortified fish sauce in improving the iron status of anemic women. DESIGN: In a randomized, double-masked study of 152 anemic (hemoglobin concentration of 81-119 g/L) women, a meal based on noodles or rice was served 6 d/wk with 10 mL fish sauce containing either 10 mg Fe as NaFeEDTA (iron-fortified group) or no added iron (control group). Concentrations of hemoglobin, serum ferritin (SF), and serum transferrin receptor (TfR) were measured at baseline and after 3 and 6 mo. RESULTS: After 6 mo, hemoglobin and SF concentrations were higher and TfR concentrations were lower in the iron-fortified group than in the control group [hemoglobin: 116.3 +/- 8.7 ( +/- SD) compared with 107.6 +/- 11.0 g/L (P < 0.0001); SF: 30.9 (95% CI: 23.4, 40.6) compared with 14.6 (11.3, 19.0) micro g/L (P = 0.0002); TfR: 7.2 (6.4, 7.9) compared with 9.0 (8.1, 9.9) mg/L (P = 0.002)]. The prevalence of iron deficiency (SF < 12 micro g/L or TfR > 8.5 mg/L) and iron deficiency anemia (iron deficiency with hemoglobin < 120 g/L) was lower in the iron-fortified group than in the control group [32.8% compared with 62.5% (P = 0.0005) and 20.3% compared with 58.3% (P < 0.0001), respectively]. CONCLUSIONS: Regular consumption of iron-fortified fish sauce significantly reduced the prevalence of iron deficiency anemia in Vietnamese women during the 6-mo intervention. Fortifying fish sauce with iron by using a water-soluble, highly bioavailable compound (NaFeEDTA) is a promising strategy for combating iron deficiency anemia in Vietnam. PMID- 12885711 TI - Infant growth and health outcomes associated with 3 compared with 6 mo of exclusive breastfeeding. AB - BACKGROUND: Opinions and recommendations about the optimal duration of exclusive breastfeeding have been strongly divided, but few published studies have provided direct evidence on the relative risks and benefits of different breastfeeding durations in recipient infants. OBJECTIVE: We examined the effects on infant growth and health of 3 compared with 6 mo of exclusive breastfeeding. DESIGN: We conducted an observational cohort study nested within a large randomized trial in Belarus by comparing 2862 infants exclusively breastfed for 3 mo (with continued mixed breastfeeding through >/= 6 mo) with 621 infants who were exclusively breastfed for >/= 6 mo. Regression to the mean, within-cluster correlation, and cluster- and individual-level confounding variables were accounted for by using multilevel regression analyses. RESULTS: From 3 to 6 mo, weight gain was slightly greater in the 3-mo group [difference: 29 g/mo (95% CI: 13, 45 g/mo)], as was length gain [difference: 1.1 mm (0.5, 1.6 mm)], but the 6-mo group had a faster length gain from 9 to 12 mo [difference: 0.9 mm/mo (0.3, 1.5 mm/mo)] and a larger head circumference at 12 mo [difference: 0.19 cm (0.07, 0.31 cm)]. A significant reduction in the incidence density of gastrointestinal infection was observed during the period from 3 to 6 mo in the 6-mo group [adjusted incidence density ratio: 0.35 (0.13, 0.96)], but no significant differences in risk of respiratory infectious outcomes or atopic eczema were apparent. CONCLUSIONS: Exclusive breastfeeding for 6 mo is associated with a lower risk of gastrointestinal infection and no demonstrable adverse health effects in the first year of life. PMID- 12885712 TI - Nutritional evaluation of protein hydrolysate formulas in healthy term infants: plasma amino acids, hematology, and trace elements. AB - BACKGROUND: Protein hydrolysate formulas are used for infants with food allergy. Most studies of such formulas focus on their effect on allergy and rarely evaluate their capacity to provide normal nutritional status. OBJECTIVE: We compared plasma aminograms, serum urea nitrogen, and trace element status in breastfed infants, infants fed hydrolysate formulas, and infants fed milk formula. DESIGN: From 6 wk to 6 mo of age, infants were breastfed or fed regular milk formula (RF), 1 of 2 casein-hydrolysate formulas (CH-1 or CH-2), or whey hydrolysate formula (WH). Anthropometric measures were taken monthly, and blood samples were collected at 6 wk and 6 mo. Plasma amino acids, serum urea nitrogen, hematologic indexes, plasma zinc, and plasma copper were analyzed. RESULTS: There were no significant differences in hemoglobin, serum transferrin receptor, copper, or zinc among groups. Serum ferritin was significantly lower in infants fed the CH formulas than in the other groups. Infants fed CH-2 had significantly higher serum urea nitrogen than did all other groups. Plasma threonine, valine, phenylalanine, methionine, and tryptophan were significantly higher in the hydrolysate formula groups than in the breastfed group. Plasma tyrosine was significantly lower in infants fed the CH formulas than in the breastfed group, whereas arginine was significantly higher in the WH group than in all other groups. Plasma proline was lower, whereas threonine and tryptophan were higher, in the WH group than in the CH groups. CONCLUSIONS: The iron status of infants fed CH formula was lower than that of all other groups. The amounts of amino acids provided by hydrolysate formulas appear excessive compared with regular formula, which is reflected by high serum urea nitrogen (CH-2) and high plasma amino acid concentrations. A reduced and more balanced amino acid content of hydrolysate formulas may be beneficial. PMID- 12885713 TI - Comparison of the efficacy of a solid ready-to-use food and a liquid, milk-based diet for the rehabilitation of severely malnourished children: a randomized trial. AB - BACKGROUND: The World Health Organization recommends a liquid, milk-based diet (F100) during the rehabilitation phase of the treatment of severe malnutrition. A dry, solid, ready-to-use food (RTUF) that can be eaten without adding water has been proposed to eliminate the risk of bacterial contamination from added water. The efficacies of RTUF and F100 have not been compared. OBJECTIVE: The objective was to compare the efficacy of RTUF and F100 in promoting weight gain in malnourished children. DESIGN: In an open-labeled, randomized trial, 70 severely malnourished Senegalese children aged 6-36 mo were randomly allocated to receive 3 meals containing either F100 (n = 35) or RTUF (n = 35) in addition to the local diet. The data from 30 children in each group were analyzed. RESULTS: The mean (+/- SD) daily energy intake in the RTUF group was 808 +/- 280 (95% CI: 703.8, 912.9) kJ x kg body wt(-1) x d(-1), and that in the F100 group was 573 +/- 201 (95% CI: 497.9, 648.7) kJ. kg body wt(-1) x d(-1) (P < 0.001). The average weight gains in the RTUF and F100 groups were 15.6 (95% CI: 13.4, 17.8) and 10.1 (95% CI: 8.7, 11.4) g x kg body wt(-1) x d(-1), respectively (P < 0.001). The difference in weight gain was greater in the most wasted children (P < 0.05). The average duration of rehabilitation was 17.3 (95% CI: 15.6, 19.0) d in the F100 group and was 13.4 (95% CI: 12.1, 14.7) d in the RTUF group (P < 0.001). CONCLUSIONS: This study indicated that RTUF can be used efficiently for the rehabilitation of severely malnourished children. PMID- 12885714 TI - Body composition and overweight prevalence in 1704 schoolchildren from 7 American Indian communities. AB - BACKGROUND: Nationwide data on obesity prevalence in American Indian communities are limited. OBJECTIVE: We describe the body composition and anthropometric characteristics of schoolchildren from 7 American Indian communities enrolled in the Pathways study, a randomized field trial evaluating a program for the primary prevention of obesity. DESIGN: A total of 1704 children in 41 schools were enrolled in the study. Basic anthropometric measurements included weight, height, and triceps and subscapular skinfold thicknesses. Percentage body fat was estimated from bioelectrical impedance and anthropometric variables with the use of an equation developed and validated for this population. RESULTS: The children's mean (+/- SD) age was 7.6 +/- 0.6 y, and their mean weight and height were 32.1 +/- 8.9 kg and 129.8 +/- 6.3 cm, respectively. Mean body mass index (BMI; in kg/m(2)) was 18.8 +/- 3.9, and mean percentage body fat was 32.6 +/- 6.8%. With the use of current Centers for Disease Control and Prevention reference values, 30.5% of girls and 26.8% of boys were above the 95th percentiles for BMI-for-age, and 21% of girls and 19.6% of boys were between the 85th and 95th percentiles. Although there was a wide range in BMI across study sites and for both sexes, the percentage of children with a BMI above the 95th percentile was consistently higher than the national averages in all communities studied and in both girls and boys. CONCLUSIONS: Overweight can be documented in a substantial number of American Indian children by the time they reach elementary school. Despite differences in the prevalence of overweight observed among communities, rates are uniformly high relative to national all-race averages. PMID- 12885715 TI - Long-chain polyunsaturated fatty acids have a positive effect on the quality of general movements of healthy term infants. AB - BACKGROUND: Whether long-chain polyunsaturated fatty acids (LCPs) play a role in the development of the young nervous system in term infants is debated. OBJECTIVE: We investigated whether supplementation of formula with LCPs for 2 mo improves the quality of general movements (GMs) in healthy term infants at 3 mo of age. DESIGN: A prospective, double-blind, randomized controlled study was conducted with 2 groups of healthy term infants: a control-formula (CF) group (n = 131) and an LCP-supplemented-formula (LF) group (n = 119). A breastfed (BF) group (n = 147) served as a reference. Information on potential confounders was collected at enrollment. Videotapes were made of the infants' spontaneous motor behavior at 3 mo of age to assess the quality of their GMs. On the basis of quality, normal GMs were classified as normal-optimal or normal-suboptimal, and abnormal GMs were classified as mildly or definitely abnormal. Attrition at 3 mo of age was 15% and nonselective. Multivariate regression analyses with adjustment for confounders were carried out to evaluate the effect of the type of feeding. RESULTS: None of the infants had definitely abnormal GMs. Infants in the CF group had mildly abnormal GMs significantly more often than did infants in the LF and BF groups (31% compared with 19% and 20%, respectively). Infants in the BF group had normal-optimal GMs more frequently than did infants in the LF and CF groups (34% compared with 18% and 21%, respectively). Logistic regression analyses confirmed these findings. CONCLUSION: Supplementation of healthy term infants with LCPs during the first 2 mo of life reduces the occurrence of mildly abnormal GMs. PMID- 12885716 TI - Effects of tuberculosis and HIV infection on whole-body protein metabolism during feeding, measured by the [15N]glycine method. AB - BACKGROUND: Tuberculosis (TB) and HIV infection are wasting diseases that frequently occur together and have severe consequences on nutritional status. OBJECTIVE: The objective was to determine the effects of TB and HIV, separately and together, on protein metabolism. DESIGN: Protein metabolism was determined in the fed state in 11 healthy control subjects, in 10 patients with HIV infection without TB or other active infection (HIV group), in 10 patients with active TB without HIV infection (TB group), and in 8 patients with HIV infection and active TB (HIVTB group) with the use of oral [(15)N]glycine and measurement of enrichment in urinary urea and ammonia. RESULTS: Whole-body protein flux and degradation were lower in the HIV group than in the control group (mean flux: 3.53 +/- 0.40 compared with 4.75 +/- 0.97 g. kg lean body mass(-1). 12 h(-1); P = 0.002). Protein flux, synthesis, and degradation were not significantly different between the control group and the TB and HIVTB groups. Net protein balance was strongly anabolic in the control, HIV, and TB groups but was neutral in the HIVTB group (P < 0.001 for comparison between groups). CONCLUSIONS: HIV infection was associated with a significant down-regulation of whole-body protein flux. TB alone was not associated with abnormal protein metabolism, but net anabolism in the fed state was impaired in the HIVTB group. PMID- 12885717 TI - Influence of nutritional status on alcoholic myopathy. AB - BACKGROUND: Muscle weakness and structural changes in striated skeletal muscle are common in persons with chronic alcoholism. OBJECTIVE: The objective of the study was to assess the role of malnutrition in the development of chronic alcoholic myopathy. DESIGN: We prospectively evaluated 146 men who reported an intake >/=100 g ethanol/d for the previous 5 y and 73 well-nourished control subjects. Alcohol consumption, energy and protein nutritional status, and deltoid muscle strength were determined. Deltoid muscle tissue specimens were taken from alcoholics and from 14 control subjects for histochemical studies and morphometric measurements of the fibers. RESULTS: Deltoid muscle strength was less in alcoholics than in control subjects (P < 0.001). Muscle strength correlated with lifetime consumption of ethanol (r = -0.56, P < 0.001), and a decrease in muscle strength was significantly greater in the presence of energy malnutrition. Using logistic regression analysis, we observed that alcoholics with muscle strength < 18 kg had the independent risk factors of an arm muscle area < 50 cm(2) (odds ratio: 5.4; 95% CI: 2.3, 12.3), consumption of > 1600 kg ethanol throughout their lives (odds ratio: 4.5; 95% CI: 2.0, 10.1), and protein malnutrition (odds ratio: 4.2; 95% CI: 1.4, 12.7). Protein malnutrition was also associated with muscle inefficiency (P < 0.001). Histologic myopathy was present in 58% of alcoholics, was related to lifetime ethanol consumption (P = 0.001), and was more severe in the presence of protein malnutrition (P = 0.01). CONCLUSION: Malnutrition is an additional developmental factor in the functional and structural muscle damage induced by chronic ethanol consumption. PMID- 12885718 TI - Relation between homocysteine concentrations and the consumption of different types of alcoholic beverages: the French Supplementation with Antioxidant Vitamins and Minerals Study. AB - BACKGROUND: Previous studies on the effects of alcohol consumption on total plasma homocysteine (tHcy) concentrations showed contradictory results. The conflicting results may derive in part from confounding by the type of alcoholic beverage consumed. OBJECTIVE: The objective of the study was to evaluate in a predominantly wine-drinking French population whether the relation between alcohol consumption and homocysteine concentrations is dependent on the type of alcoholic beverage consumed. DESIGN: In 1996, a cross-sectional study measuring tHcy and red blood cell folate concentrations was conducted in 1196 middle-aged women and men from the French Supplementation with Antioxidant Vitamins and Minerals Study. Intakes of alcohol, energy, coffee, and B vitamins were assessed by 6 separate 24-h dietary records from the previous year. RESULTS: tHcy concentrations were positively associated with wine intake (P = 0.01) in the women and with beer intake in the men (P = 0.002). No association with the consumption of spirits was observed. The association between beer consumption and tHcy concentrations in the men was modified by the consumption of wine; the association was positive in wine drinkers, whereas an inverse trend was seen in those who drank no wine. CONCLUSION: Wine consumption may increase tHcy concentrations, whereas beer consumption seems to have no effect (or even an inverse effect) on tHcy. PMID- 12885719 TI - Alcohol consumption in relation to risk of cholecystectomy in women. AB - BACKGROUND: Alcohol consumption has been linked to a lower risk of gallstone disease. However, the magnitude of the association is uncertain, and little is known about the relation of alcohol consumption patterns and individual types of alcoholic beverages to gallstone disease risk. OBJECTIVE: We prospectively examined the association between alcohol intake and cholecystectomy, a surrogate for symptomatic gallstone disease, in a large cohort of women. DESIGN: Women from the Nurses' Health Study who had no history of gallstone disease in 1980 (n = 80,898) were followed for 20 y. Alcohol consumption, which was measured every 2-4 y by food-frequency questionnaires, was used to predict subsequent cholecystectomy through multivariate analysis. RESULTS: We ascertained 7831 cases of cholecystectomy. Relative to subjects who had no alcohol intake, subjects who had alcohol intakes of 0.1-4.9, 5.0-14.9, 15.0-29.9, 30.0-49.9, and >/=50.0 g/d had multivariate relative risks of cholecystectomy of 0.95, 0.86, 0.80, 0.67, and 0.62 (95% CI: 0.49, 0.79), respectively. Relative to subjects who never consumed alcohol, subjects who consumed alcohol 1-2, 3-4, 5-6, and 7 d/wk had multivariate relative risks of cholecystectomy of 0.94, 0.88, 0.87, and 0.73 (0.63, 0.84), respectively. All alcoholic beverage types were inversely associated with cholecystectomy risk, independent of consumption patterns (for quantity of alcohol consumed, P = 0.04, 0.001, and 0.003 for wine, beer, and liquor, respectively; for frequency of alcohol consumption, P = 0.01, 0.07, and <0.0001 for wine, beer, and liquor, respectively). CONCLUSIONS: The intake of all alcoholic beverage types is inversely associated with the risk of cholecystectomy. Recommendations regarding the benefit of consuming moderate quantities of alcohol should be weighed against the potential health hazards. PMID- 12885721 TI - Body mass index and body fat. PMID- 12885723 TI - Alternate healthy eating index. PMID- 12885724 TI - Bone mineral content, not bone mineral density, is the correct bone measure for growth studies. PMID- 12885727 TI - Plasminogen activator inhibitor-1 and the calculus of mortality after myocardial infarction. PMID- 12885728 TI - Bone marrow: a fountain of vascular youth? PMID- 12885729 TI - Proteomics in cardiovascular biology and medicine. PMID- 12885730 TI - Vascular radiation in a drug-eluting stent world: it's not over till it's over. PMID- 12885731 TI - Coronary brachytherapy in the drug-eluting stent era: don't bury it alive. PMID- 12885732 TI - Vascular brachytherapy boon or bust? PMID- 12885733 TI - Clinical use of inotropic therapy for heart failure: looking backward or forward? Part II: chronic inotropic therapy. PMID- 12885734 TI - Images in cardiovascular medicine. Magnetic resonance imaging of impending left ventricular rupture after acute myocardial infarction. PMID- 12885735 TI - Images in cardiovascular medicine. Diagnosis of myocarditis by cardiac tissue velocity imaging in an olympic athlete. PMID- 12885736 TI - Variable titin-based stiffness adjustment in heart disease. PMID- 12885737 TI - Ascertainment strategies and genotype:phenotype correlations in hypertrophic cardiomyopathy. PMID- 12885738 TI - Potential antiapoptotic activity of aldosterone antagonists in postinfarction remodeling. PMID- 12885739 TI - Effect of epicardial or biventricular pacing to prolong QT interval and increase transmural dispersion of repolarization. PMID- 12885740 TI - Low triiodothyronine and cardiovascular disease. PMID- 12885741 TI - Aging of progenitor cells contributes to atherosclerosis. PMID- 12885742 TI - Prognostic value of lead aVR in patients with a first non-ST-segment elevation acute myocardial infarction. AB - BACKGROUND: ST-segment elevation in lead aVR has been associated with severe coronary artery lesions in patients with acute coronary syndromes, but the prognostic significance of this finding is unknown. METHODS AND RESULTS: We analyzed the initial ECG in 775 consecutive patients admitted to our center with a first acute myocardial infarction without ST-segment elevation in leads other than aVR or V1. The rates of in-hospital death in patients without (n=525) and with 0.05 to 0.1 mV (n=116) or > or =0.1 mV (n=134) of ST-segment elevation in lead aVR were 1.3%, 8.6%, and 19.4%, respectively (P<0.001). After adjustment for the baseline clinical predictors and for ST-segment depression on admission, the odds ratios for death in the last 2 groups were, respectively, 4.2 (95% CI, 1.5 to 12.2) and 6.6 (95% CI, 2.5 to 17.6). The rates of recurrent ischemic events and heart failure during hospital stay also increased in a stepwise fashion among the groups, whereas creatine kinase-MB levels were similar. Among the 437 patients that were catheterized within 6 months, the prevalence of left main or 3 vessel coronary artery disease in the 3 groups was 22.0%, 42.6%, and 66.3%, respectively (P<0.001). CONCLUSIONS: Lead aVR contains important short-term prognostic information in patients with a first non-ST-segment elevation acute myocardial infarction. Because the poorer outcome predicted by ST-segment elevation in lead aVR seems to be related to a more severe coronary artery disease, an early invasive approach might be especially beneficial in patients presenting with this finding. PMID- 12885743 TI - Do hospitals and surgeons with higher coronary artery bypass graft surgery volumes still have lower risk-adjusted mortality rates? AB - BACKGROUND: Studies that are the basis of recommended volume thresholds for CABG surgery are outdated and not reflective of recent advances in the field. This study examines both hospital and surgeon volume-mortality relations for CABG surgery through the use of a population-based clinical data set. METHODS AND RESULTS: Data from New York's clinical CABG surgery registry from 1997 to 1999 (total number of procedures, 57 150) were used to examine the individual and combined impact of annual hospital volume and annual surgeon volume on in hospital mortality rates after adjusting for differences in severity of illness. Significantly lower risk-adjusted mortality rates occurred above all annual hospital volume thresholds between 200 and 800 and above all surgeon volume thresholds between 50 and 200. The number needed to treat (NNT) at higher-volume providers to avoid a death was minimized for a hospital threshold volume of 100 (NNT=50) and a surgeon threshold volume of 50 (NNT=118). The risk-adjusted mortality rate (RAMR) for patients undergoing surgery performed by surgeons with volumes of > or =125 in hospitals with volumes of > or =600 was 1.89%. The RAMR was significantly higher (2.67%) for patients undergoing surgery performed by surgeons with volumes of <125 in hospitals with volumes of <600. CONCLUSIONS: Higher-volume surgeons and hospitals continue to have lower risk-adjusted mortality rates, and patients undergoing surgery performed by higher-volume surgeons in higher-volume hospitals have the lowest mortality rates. PMID- 12885744 TI - Percutaneous balloon valvotomy in pulmonary atresia with intact ventricular septum: impact on patient care. AB - BACKGROUND: Pulmonary atresia with intact ventricular septum (PA-IVS) is a rare congenital lesion with high mortality. Therapy was exclusively surgical until recently, when the use of radiofrequency-assisted perforation of the atretic valve was introduced as a treatment option. This study analyzes the outcomes and morphological changes to right heart structures after percutaneous perforation and balloon dilation of the atretic valve. METHODS AND RESULTS: Between April 1992 and August 2000, 30 patients with PA-IVS underwent attempted percutaneous valve perforation and balloon dilation of the pulmonary valve. Longitudinal echocardiographic measurements of the tricuspid valve diameter, right ventricular length and area were recorded. Z scores were calculated according to published formulas. Perforation was achieved in 27 patients. In 14 patients a modified Blalock-Taussig shunt was performed between 2 and 24 days after valve dilation. There were 3 early and 2 late deaths. Among the survivors (follow-up time of 1 to 87 months), 16 patients had a biventricular circulation, 3 a 1(1/2)-ventricle circulation, and 1 a Fontan operation. Four patients are awaiting further palliation. There was no significant change of the tricuspid valve Z score or right ventricular length Z score with time. CONCLUSIONS: Percutaneous balloon valvotomy is an effective treatment strategy for patients with PA-IVS provided that there is a patent infundibulum and a lack of a right ventricle-dependent coronary circulation. Despite the observation that right heart growth does not increase with body growth in early follow-up, it appears adequate to maintain a biventricular circulation in many patients. PMID- 12885745 TI - Short-term statin therapy improves cardiac function and symptoms in patients with idiopathic dilated cardiomyopathy. AB - BACKGROUND: Chronic heart failure is associated with inflammation and neurohormonal imbalance. The 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) reductase inhibitors, or statins, exert anti-inflammatory and vascular protective effects. We hypothesized that short-term statin therapy may have beneficial effects in patients with nonischemic heart failure. METHODS AND RESULTS: Sixty-three patients with symptomatic, nonischemic, dilated cardiomyopathy were randomly divided into 2 groups. One group received simvastatin (n=24), and the other group received placebo (n=27). The initial dose of simvastatin was 5 mg/d, which was increased to 10 mg/d after 4 weeks. After 14 weeks, patients receiving simvastatin exhibited a modest reduction in serum cholesterol level compared with patients receiving placebo (130+/-13 versus 148+/-18, P<0.05). Patients treated with simvastatin had a lower New York Heart Association functional class compared with patients receiving placebo (2.04+/-0.06 versus 2.32+/-0.05, P<0.01). This corresponded to improved left ventricular ejection fraction in the simvastatin group (34+/-3 to 41+/-4%, P<0.05) but not in the placebo group. Furthermore, plasma concentrations of tumor necrosis factor-alpha, interleukin-6, and brain natriuretic peptide were significantly lower in the simvastatin group compared with the placebo group. CONCLUSIONS: Short-term statin therapy improves cardiac function, neurohormonal imbalance, and symptoms associated with idiopathic dilated cardiomyopathy. These findings suggest that statins may have therapeutic benefits in patients with heart failure irrespective of serum cholesterol levels or atherosclerotic heart disease. PMID- 12885746 TI - Characterization of endocardial electrophysiological substrate in patients with nonischemic cardiomyopathy and monomorphic ventricular tachycardia. AB - BACKGROUND: Although catheter mapping has been used to define the endocardial electrogram characteristics in patients with ventricular tachycardia (VT) and coronary disease, characterization of the electrophysiological substrate in patients with VT and nonischemic cardiomyopathy is limited. METHODS AND RESULTS: Left ventricular endocardial electroanatomical mapping was performed in 19 patients with nonischemic cardiomyopathy and monomorphic VT with an average of 178+/-83 sites per chamber mapped. Abnormal bipolar electrogram was defined as endocardial voltage signal amplitude of <1.8 mV. The extent and location of abnormal endocardium was estimated by measuring areas of abnormal electrogram recordings from 3D voltage maps. The origin of VT was approximated by identifying sites of entrainment with concealed fusion or early presystolic activity and/or by pace mapping. Abnormal electrograms were recorded over a 41+/-28 cm2 area that represented 20+/-12% of total endocardial surface. The majority of patients (14/19 patients) had only a modest area (<25%) of endocardial abnormality. All patients had abnormal low-voltage endocardial areas located near the ventricular base in the perivalvular region. There were 3+/-1 VT morphologies per patient. The majority (88%) of the 57 mapped VTs originated from the ventricular base, corresponding to regions with abnormal endocardial electrograms. CONCLUSIONS: Electroanatomical mapping in patients with monomorphic VT and nonischemic cardiomyopathy typically demonstrates a modest-sized basal area of endocardial electrogram abnormalities. The VT site of origin corresponds to these basal electrogram abnormalities. These findings have important implications regarding strategies for VT ablation in this setting. PMID- 12885747 TI - Regression of electrocardiographic left ventricular hypertrophy by losartan versus atenolol: The Losartan Intervention for Endpoint reduction in Hypertension (LIFE) Study. AB - BACKGROUND: Electrocardiographic left ventricular hypertrophy (LVH) predicts cardiovascular morbidity and mortality, and regression of ECG LVH may predict improved prognosis in hypertensive patients. However, uncertainty persists as to how best to regress ECG LVH. METHODS AND RESULTS: Regression of ECG LVH with losartan versus atenolol therapy was assessed in 9193 hypertensive patients with ECG LVH by Sokolow-Lyon voltage or Cornell voltage-duration product criteria enrolled in the Losartan Intervention For Endpoint Reduction in Hypertension (LIFE) Study. Patients had ECGs at study baseline and after 6 months, 1, 2, 3, 4, and 5 years of blinded losartan-based or atenolol-based therapy. After 6 months' follow-up, adjusting for baseline ECG LVH levels, baseline and in-treatment systolic and diastolic pressures, and for diuretic therapy, losartan-based therapy was associated with greater regression of both Cornell product (adjusted means, -200 versus -69 mm. ms, P<0.001) and Sokolow-Lyon voltage (-2.5 versus 0.7 mm, P<0.001) than was atenolol-based therapy. Greater regression of ECG LVH persisted at each subsequent annual evaluation in the losartan-treated group, with between 140 and 164 mm. ms greater mean reductions in Cornell product and from 1.7 to 2.2 mm greater mean reductions in Sokolow-Lyon voltage (all P<0.001). The effect of losartan was consistent across subgroups defined by gender, age, ethnicity, and diabetes. CONCLUSIONS: After adjusting for baseline and in treatment blood pressure and baseline severity of ECG LVH, losartan-based antihypertensive therapy resulted in greater regression of ECG LVH by Cornell voltage-duration product and Sokolow-Lyon voltage criteria than did atenolol based therapy. These findings support the value of angiotensin receptor blockade with losartan for reversing ECG LVH. PMID- 12885748 TI - Effect of 7-year infancy-onset dietary intervention on serum lipoproteins and lipoprotein subclasses in healthy children in the prospective, randomized Special Turku Coronary Risk Factor Intervention Project for Children (STRIP) study. AB - BACKGROUND: We previously showed that low-saturated-fat dietary intervention from infancy until 5 years of age safely and effectively reduced serum cholesterol concentration. We now report how such intervention influenced serum lipids, LDL particle size, and HDL subfractions in children when they reached the age of 7 years. METHODS AND RESULTS: Healthy 7-month-old infants (n=1062) were randomized to the intervention (n=540) and control (n=522) groups. Each year, two individualized counseling sessions were organized to the intervention families. Serum lipid values were measured annually. The intervention boys had 0.20 to 0.39 mmol/L lower serum cholesterol values than the control boys throughout the follow up (always P<0.05), but the values of the intervention and control girls did not differ. The LDL particle sizes and HDL subfractions were determined in a random subgroup of 96 intervention and 101 control children at the age of 7 years. The mean particle diameter of major LDL peak was 262.6 A in the intervention boys and 258.5 A in the control boys (P=0.05), and 259.2 A in the intervention girls and 261.3 A in the control girls (P=0.30). HDL2 and HDL3 cholesterol concentrations did not differ between the intervention and control children or between the two genders. CONCLUSIONS: The 7-year intervention favorably influenced not only the serum total and LDL cholesterol concentrations but also the LDL particle size in boys. LDL particle size remained unchanged in girls, as did HDL2 and HDL3 concentrations in both genders. PMID- 12885749 TI - Increasing trends in hospitalization for atrial fibrillation in the United States, 1985 through 1999: implications for primary prevention. AB - BACKGROUND: Atrial fibrillation, the most common sustained disturbance of heart rhythm, is associated with a 5-fold increase in the incidence of ischemic stroke. METHODS AND RESULTS: The National Hospital Discharge Survey was used to estimate the annual number and prevalence of hospitalizations with atrial fibrillation among men and women 35 years of age or older. From 1985 through 1999, hospitalizations increased from 154 086 to 376 487 for a first-listed diagnosis and from 787 750 to 2 283 673 for any diagnosis. Prevalence was higher among successive age groups. Age-standardized prevalence was consistently higher among men than women. In 1999, essential hypertension, ischemic heart disease, congestive heart failure, and diabetes were prominent coexisting conditions. The number of male patients discharged home decreased from 77% to 63%, whereas the number of discharges to long-term care increased from 9% to 15%; the corresponding values for women were 72% to 56% and 15% to 23%. A slight increase in discharges to short-term care was indicated, whereas no trends were noted for in-hospital mortality. CONCLUSIONS: Hospitalizations for atrial fibrillation have increased dramatically (2- to 3-fold) in recent years. The public health burden of atrial fibrillation is enormous and expected to continue to increase over the next decades. Primary prevention of atrial fibrillation must be recognized and pursued as a complementary management strategy for reducing cardiovascular morbidity and mortality. PMID- 12885750 TI - Norepinephrine precursor therapy in neurogenic orthostatic hypotension. AB - BACKGROUND: In patients with neurogenic orthostatic hypotension (NOH), the availability of the sympathetic neurotransmitter norepinephrine (NE) in the synaptic cleft is insufficient to maintain blood pressure while in the standing posture. METHODS AND RESULTS: We determined the effect of oral administration of the synthetic amino acid L-threo-3,4-dihydroxyphenylserine (L-DOPS), which is decarboxylated to NE by the enzyme L-aromatic amino acid decarboxylase (L-AADC) in neural and nonneural tissue, on blood pressure and orthostatic tolerance in 19 patients with severe NOH (8 with pure autonomic failure and 11 with multiple system atrophy). A single-blind dose-titration study determined the most appropriate dose for each patient. Patients were then enrolled in a double-blind, placebo-controlled, crossover trial. L-DOPS significantly raised mean blood pressure both supine (from 101+/-4 to 141+/-5 mm Hg) and standing (from 60+/-4 to 100+/-6 mm Hg) for several hours and improved orthostatic tolerance in all patients. After L-DOPS, blood pressure increases were closely associated with increases in plasma NE levels. Oral administration of carbidopa, which inhibits L AADC outside the blood-brain barrier, blunted both the increase in plasma NE and the pressor response to L-DOPS in all patients CONCLUSIONS: Acute administration of L-DOPS increases blood pressure and improves orthostatic tolerance in patients with NOH. The pressor effect results from conversion of L-DOPS to NE outside the central nervous system. PMID- 12885751 TI - Decline in out-of-hospital coronary heart disease deaths has contributed the main part to the overall decline in coronary heart disease mortality rates among persons 35 to 64 years of age in Finland: the FINAMI study. AB - BACKGROUND: Out-of-hospital deaths constitute the majority of all coronary heart disease (CHD) deaths and are therefore of considerable public health significance. METHODS AND RESULTS: We used population-based myocardial infarction register data to examine trends in out-of-hospital CHD deaths in Finland during 1983 to 1997. We included in out-of-hospital deaths also deaths in the emergency room and all deaths within 1 hour after the onset of symptoms. Altogether, 3494 such events were included in the analyses. The proportion of out-of-hospital deaths of all CHD deaths depended on age and gender. In the age group 35 to 64 years, it was 73% among men and 60% among women. These proportions did not change during the study. The annual average decline in the age-standardized out-of hospital CHD death rate was 6.1% (95% CI, -7.3, -5.0%) among men and 7.0% (-10.0, -4.0%) among women. These declines contributed among men 70% and among women 58% to the overall decline in CHD mortality rate. In all, 58% of the male and 52% of the female victims of out-of-hospital CHD death had a history of symptomatic CHD. Among men with a prior history of myocardial infarction, the annual average decline in out-of-hospital CHD deaths was 5.3% (-7.2, -3.2%), and among men without such history the decline was 2.9% (-4.4, -1.5%). Among women, the corresponding changes were -7.8% (-14.2, -1.5%) and -4.5% (-8.0, -1.0%). CONCLUSIONS: The decline in out-of-hospital CHD deaths has contributed the main part to the overall decline in CHD mortality rates among persons 35 to 64 years of age in Finland. PMID- 12885752 TI - Ionic remodeling of sinoatrial node cells by heart failure. AB - BACKGROUND: In animal models of heart failure (HF), heart rate decreases as the result of an increase in intrinsic cycle length of the sinoatrial node (SAN). In this study, we evaluate the HF-induced remodeling of membrane potentials and currents in SAN cells. METHODS AND RESULTS: SAN cells were isolated from control rabbits and rabbits with volume and pressure overload-induced HF and patch clamped to measure their electrophysiological properties. HF cells were not hypertrophied (capacitance, mean+/-SEM, 52+/-3 versus 50+/-4 pF in control). HF increased intrinsic cycle length by 15% and decreased diastolic depolarization rate by 30%, whereas other action potential parameters were unaltered. In HF, the hyperpolarization-activated "pacemaker" current (If) and slow component of the delayed rectifier current (IKs) were reduced by 40% and 20%, respectively, without changes in voltage dependence or kinetics. T-type and L-type calcium current, rapid and ultrarapid delayed rectifier current, transient outward currents, and sodium-calcium exchange current were unaltered. CONCLUSIONS: In single SAN cells of rabbits with HF, intrinsic cycle length is increased as the result of a decreased diastolic depolarization rate rather than a change in action potential duration. HF reduced both If and IKs density. Since IKs plays a limited role in pacemaker activity, the HF-induced decrease in heart rate is attributable to remodeling of If. PMID- 12885753 TI - A20 protects from CD40-CD40 ligand-mediated endothelial cell activation and apoptosis. AB - BACKGROUND: CD40/CD40 ligand (CD40L) signaling is a potent activator of endothelial cells (ECs) and promoter of atherosclerosis. In this study, we investigate whether A20 (a gene we have shown to be antiinflammatory and antiapoptotic in ECs) can protect from CD40/CD40L-mediated EC activation. METHODS AND RESULTS: Overexpression of CD40, in a transient transfection system, activates the transcription factor NF-kappaB and upregulates IkappaBalpha, E selectin, and tissue factor (TF) reporter activity. Coexpression of A20 inhibits NF-kappaB and upregulation of IkappaBalpha and E-Selectin but not TF, suggesting that CD40 induces TF in a non-NF-kappaB-dependent manner. In human coronary artery ECs (HCAECs), adenovirus-mediated overexpression of A20 blocks physiological, CD40-induced activation of NF-kappaB, upstream of IkappaBalpha degradation (Western blot) and subsequently upregulation of ICAM-1, VCAM-1, and E selectin (flow cytometry). Although A20 does not block TF transcription its expression in HCAECs inhibits TF induction (colorimetric assay and RT-PCR) by blunting CD40 upregulation. We demonstrate that CD40 signaling induces apoptosis in a proinflammatory microenvironment. A20 overexpression protects from CD40 mediated EC apoptosis (DNA content analysis and trypan blue exclusion). We also demonstrate that signaling through CD40L activates NF-kappaB and induces apoptosis in ECs, both of which are inhibited by A20 overexpression. CONCLUSIONS: A20 works at multiple levels to protect ECs from CD40/CD40L mediated activation and apoptosis. A20-based therapy could be beneficial for the treatment of vascular diseases such as atherosclerosis and transplant-associated vasculopathy. PMID- 12885754 TI - Prophylactic efficacy of topical temporin A and RNAIII-inhibiting peptide in a subcutaneous rat Pouch model of graft infection attributable to staphylococci with intermediate resistance to glycopeptides. AB - BACKGROUND: Bacteria that adhere to implanted medical devices play an important role in industry and in modern medicine. Staphylococci are among the most common pathogens that cause biomaterial infections. Vascular prosthetic graft infection is one of the most feared complications that the vascular surgeon treats, frequently resulting in prolonged hospitalization, organ failure, amputation, and death. A rat model was used to investigate the topical efficacies of temporin A and the quorum-sensing inhibitor RNAIII-inhibiting protein (RIP) as prophylactic agents of vascular prosthetic graft infections caused by Staphylococcus aureus and Staphylococcus epidermidis with intermediate resistance to glycopeptides. METHODS AND RESULTS: Graft infections were established in the back subcutaneous tissue of adult male Wistar rats by implantation of Dacron prostheses 1 cm2 followed by topical inoculation with 2x10(7) colony-forming units of bacterial strains. The study included, for each staphylococcal strain, a control group (no graft contamination), a contaminated group that did not receive antibiotic prophylaxis, and 6 contaminated groups that received grafts soaked with temporin A, RIP, rifampin, temporin A plus RIP, RIP plus rifampin, or temporin A plus RIP. The infection was evaluated by quantitative agar culture. When tested alone, temporin A and RIP showed comparable efficacies, and their efficacies were significantly higher than that of rifampin against both strains. All combinations showed efficacies significantly higher than that of each single compound. The combinations of temporin A and RIP exerted the strongest antistaphylococcal efficacies, eliminating infection by 100%. CONCLUSIONS: The results of the present study make these molecules potentially useful for antimicrobial chemoprophylaxis in vascular surgery. PMID- 12885755 TI - Disruption of leptin signaling contributes to cardiac hypertrophy independently of body weight in mice. AB - BACKGROUND: Whether left ventricular hypertrophy (LVH) in obesity results from increased hemodynamic load or altered neurohormonal signaling remains controversial. Dysregulation of leptin, a neurohormone essential to energy homeostasis, is implicated in the pathogenesis of obesity. Because leptin has cardiovascular bioactivity, we hypothesized that disruption of leptin signaling mediates the development of obesity-associated LVH. METHODS AND RESULTS: We measured left ventricular (LV) wall thickness and LV mass with echocardiography in mice lacking leptin (ob/ob, n=15) or functional receptor (db/db, n=10) and controls at 2, 4, and 6 months of age. None of the mice had LVH at 2 months. Progressive obesity developed in ob/ob and db/db mice. At 6 months, LVH occurred in ob/ob and db/db compared with controls. We observed corresponding myocyte hypertrophy by light microscopy. To separate the direct contribution of leptin deficiency from mechanical effects of obesity, we induced weight loss in 6- to 8 month-old ob/ob mice either by leptin infusion or caloric restriction. Mice in both groups lost similar weight compared with placebo-treated controls. Leptin infusion completely reversed the increase in wall thickness with partial resolution of myocyte hypertrophy, whereas calorie-restricted mice had no decrease in wall thickness and a lesser change in myocyte size. CONCLUSIONS: Together these data show that the effect of leptin on LV remodeling is not attributable to weight loss alone, indicating that leptin has antihypertrophic effects on the heart, either directly or through a leptin-regulated neurohumoral pathway. Disruption of leptin signaling may represent a novel mechanism in LVH and related cardiovascular disorders. PMID- 12885756 TI - Subtype-selective antagonists of lysophosphatidic Acid receptors inhibit platelet activation triggered by the lipid core of atherosclerotic plaques. AB - BACKGROUND: Lysophosphatidic acid (LPA) is a platelet-activating component of mildly oxidized LDL (mox-LDL) and lipids isolated from human atherosclerotic plaques. Specific antagonists of platelet LPA receptors could be useful inhibitors of thrombus formation in patients with cardiovascular disease. METHODS AND RESULTS: Short-chain analogs of phosphatidic acid (PA) were examined for their effect on two initial platelet responses, platelet shape change and Ca2+ mobilization. Dioctylglycerol pyrophosphate [DGPP(8:0)] and dioctylphosphatidic acid [PA(8:0)], recently described selective antagonists of the LPA1 and LPA3 receptors, inhibited platelet activation evoked by LPA but not by other platelet stimuli. DGPP(8:0) was more potent than PA(8:0). DGPP(8:0) also inhibited platelet shape change induced by mox-LDL and lipid extracts from human atherosclerotic plaques. Notably, we demonstrate for the first time that the lipid-rich core isolated from soft plaques was able to directly induce shape change. This effect was completely abrogated by prior incubation of platelets with DGPP(8:0). Moreover, coapplication of the lipid-rich core or LPA together with subthreshold concentrations of ADP or epinephrine synergistically induced platelet aggregation; this effect was inhibited by DGPP(8:0). Analysis by liquid chromatography-mass spectrometry revealed the presence of LPA alkyl- and acyl molecular species with high platelet-activating potency (16:0-alkyl-LPA, 20:4 acyl-LPA). CONCLUSIONS: LPA molecules present in the core region of atherosclerotic plaques trigger rapid platelet activation through the stimulation of LPA1 and LPA3 receptors. Antagonists of platelet LPA receptors might provide a new strategy to prevent thrombus formation in patients with cardiovascular diseases. PMID- 12885757 TI - Impairment of thermoregulatory control of skin sympathetic nerve traffic in the elderly. AB - BACKGROUND: Human aging is characterized by a marked increase in muscle sympathetic nerve traffic (MSNA). No information exists, however, on the effects of aging on skin sympathetic nerve traffic (SSNA) and on its reflex modulation by thermoregulatory mechanisms. METHODS AND RESULTS: In 13 young, 11 middle-aged, and 12 elderly healthy subjects, we measured arterial blood pressure (Finapres), skin temperature (thermocouples), and resting MSNA and SSNA (microneurography). Measurements also included the SSNA responses to (1) an acute increase and reduction (+/-8 degrees C) in room temperature, each lasting 45 minutes and (2) an acoustic stimulus capable to trigger an emotional arousal. Although resting MSNA was progressively and significantly (P<0.05) increased from young to middle aged and elderly groups, SSNA was significantly (P<0.05) reduced in the latter compared with the former 2 groups. Cold exposure induced a SSNA increase that was significantly (P<0.01) smaller in the elderly than in young and middle-aged subjects. Conversely, heat exposure induced a SSNA reduction that was significantly (P<0.05) smaller in elderly than in young and middle-aged subjects. Compared with SSNA in young individuals, the SSNA change from cold to warm temperature was reduced by 61% in the elderly group. This was not the case, however, for the SSNA responses to the arousal stimulus, which were superimposable in the 3 groups. CONCLUSIONS: These data provide the first demonstration of a dichotomy in the MSNA and SSNA responses to aging. They also show that aging markedly impairs thermoregulatory control of SSNA and that this impairment might participate at the age-related SSNA decrease. PMID- 12885758 TI - Identification of a putative pathway for the muscle homing of stem cells in a muscular dystrophy model. AB - Attempts to repair muscle damage in Duchenne muscular dystrophy (DMD) by transplanting skeletal myoblasts directly into muscles are faced with the problem of the limited migration of these cells in the muscles. The delivery of myogenic stem cells to the sites of muscle lesions via the systemic circulation is a potential alternative approach to treat this disease. Muscle-derived stem cells (MDSCs) were obtained by a MACS(R) multisort method. Clones of MDSCs, which were Sca-1+/CD34-/L-selectin+, were found to adhere firmly to the endothelium of mdx dystrophic muscles after i.v. or i.m. injections. The subpopulation of Sca 1+/CD34- MDSCs expressing L-selectin was called homing MDSCs (HMDSCs). Treatment of HMDSCs with antibodies against L-selectin prevented adhesion to the muscle endothelium. Importantly, we found that vascular endothelium from striate muscle of young mdx mice expresses mucosal addressin cell adhesion molecule-1 (MAdCAM 1), a ligand for L-selectin. Our results showed for the first time that the expression of the adhesion molecule L-selectin is important for muscle homing of MDSCs. This discovery will aid in the improvement of a potential therapy for muscular dystrophy based on the systemic delivery of MDSCs. PMID- 12885759 TI - Regulation of cell death in mitotic neural progenitor cells by asymmetric distribution of prostate apoptosis response 4 (PAR-4) and simultaneous elevation of endogenous ceramide. AB - Cell death and survival of neural progenitor (NP) cells are determined by signals that are largely unknown. We have analyzed pro-apoptotic signaling in individual NP cells that have been derived from mouse embryonic stem cells. NP formation was concomitant with elevated apoptosis and increased expression of ceramide and prostate apoptosis response 4 (PAR-4). Morpholino oligonucleotide-mediated antisense knockdown of PAR-4 or inhibition of ceramide biosynthesis reduced stem cell apoptosis, whereas PAR-4 overexpression and treatment with ceramide analogs elevated apoptosis. Apoptotic cells also stained for proliferating cell nuclear antigen (a nuclear mitosis marker protein), but not for nestin (a marker for NP cells). In mitotic cells, asymmetric distribution of PAR-4 and nestin resulted in one nestin(-)/PAR-4(+) daughter cell, in which ceramide elevation induced apoptosis. The other cell was nestin(+), but PAR-4(-), and was not apoptotic. Asymmetric distribution of PAR-4 and simultaneous elevation of endogenous ceramide provides a possible mechanism underlying asymmetric differentiation and apoptosis of neuronal stem cells in the developing brain. PMID- 12885760 TI - The ER v-SNAREs are required for GPI-anchored protein sorting from other secretory proteins upon exit from the ER. AB - Glycosylphosphatidylinositol (GPI)-anchored proteins exit the ER in distinct vesicles from other secretory proteins, and this sorting event requires the Rab GTPase Ypt1p, tethering factors Uso1p, and the conserved oligomeric Golgi complex. Here we show that proper sorting depended on the vSNAREs, Bos1p, Bet1p, and Sec22p. However, the t-SNARE Sed5p was not required for protein sorting upon ER exit. Moreover, the sorting defect observed in vitro with bos1-1 extracts was also observed in vivo and was visualized by EM. Finally, transport and maturation of the GPI-anchored protein Gas1p was specifically affected in a bos1-1 mutant at semirestrictive temperature. Therefore, we propose that v-SNAREs are part of the cargo protein sorting machinery upon exit from the ER and that a correct sorting process is necessary for proper maturation of GPI-anchored proteins. PMID- 12885761 TI - Importin beta contains a COOH-terminal nucleoporin binding region important for nuclear transport. AB - Proteins containing a classical NLS are transported into the nucleus by the import receptor importin beta, which binds to cargoes via the adaptor importin alpha. The import complex is translocated through the nuclear pore complex by interactions of importin beta with a series of nucleoporins. Previous studies have defined a nucleoporin binding region in the NH2-terminal half of importin beta. Here we report the identification of a second nucleoporin binding region in its COOH-terminal half. Although the affinity of the COOH-terminal region for nucleoporins is dramatically weaker than that of the NH2-terminal region, sets of mutations that perturb the nucleoporin binding of either region reduce the nuclear import activity of importin beta to a similar extent ( approximately 50%). An importin beta mutant with a combination of mutations in the NH2- and COOH-terminal regions is completely inactive for nuclear import. Thus, importin beta possesses two nucleoporin binding sites, both of which are important for its nuclear import function. PMID- 12885762 TI - Regulation of the expression and processing of caspase-12. AB - Phylogenetic analysis clusters caspase-12 with the inflammatory caspases 1 and 11. We analyzed the expression of caspase-12 in mouse embryos, adult organs, and different cell types and tested the effect of interferons (IFNs) and other proinflammatory stimuli. Constitutive expression of the caspase-12 protein was restricted to certain cell types, such as epithelial cells, primary fibroblasts, and L929 fibrosarcoma cells. In fibroblasts and B16/B16 melanoma cells, caspase 12 expression is stimulated by IFN-gamma but not by IFN-alpha or -beta. The effect is increased further when IFN-gamma is combined with TNF, lipopolysaccharide (LPS), or dsRNA. These stimuli also induce caspase-1 and -11 but inhibit the expression of caspase-3 and -9. In contrast to caspase-1 and -11, no caspase-12 protein was detected in macrophages in any of these treatments. Transient overexpression of full-length caspase-12 leads to proteolytic processing of the enzyme and apoptosis. Similar processing occurs in TNF-, LPS-, Fas ligand-, and thapsigargin (Tg)-induced apoptosis. However, B16/B16 melanoma cells die when treated with the ER stress-inducing agent Tg whether they express caspase-12 or not. PMID- 12885763 TI - Infectious HIV-1 assembles in late endosomes in primary macrophages. AB - Although human immunodeficiency virus type 1 (HIV-1) is generally thought to assemble at the plasma membrane of infected cells, virions have been observed in intracellular compartments in macrophages. Here, we investigated virus assembly in HIV-1-infected primary human monocyte-derived macrophages (MDM). Electron microscopy of cryosections showed virus particles, identified by their morphology and positive labeling with antibodies to the viral p17, p24, and envelope proteins, in intracellular vacuoles. Immunolabeling demonstrated that these compartments contained the late endosomal marker CD63, which was enriched on vesicles within these structures and incorporated into the envelope of budding virions. The virus-containing vacuoles were also labeled with antibodies against LAMP-1, CD81, and CD82, which were also incorporated into the viral envelope. To assess the cellular source of infectious viruses derived from MDM, virus containing media from infected cells were precipitated with specific antibodies. Only antibodies against antigens found in late endosomes precipitated infectious virus, whereas antibodies against proteins located primarily on the cell surface did not. Our data indicate that most of the infectious HIV produced by primary macrophages is assembled on late endocytic membranes and acquires antigens characteristic of this compartment. This notion has significant implications for understanding the biology of HIV and its cell-cell transmission. PMID- 12885764 TI - Lipid rafts make for slippery platforms. AB - What's in a raft? Although cell membranes are certainly not homogeneous mixtures of lipids and proteins, almost all aspects of lipid rafts-how to define them, their size, composition, lifetime, and biological relevance-remain controversial. The answers will shape our views of signaling and of membrane dynamics. PMID- 12885765 TI - Identification of a COOH-terminal segment involved in maturation and stability of human ether-a-go-go-related gene potassium channels. AB - Mutations in the potassium channel encoded by the human ether-a-go-go-related gene (HERG) have been linked to the congenital long QT syndrome (LQTS), a cardiac disease associated with an increased preponderance of ventricular arrhythmias and sudden death. The COOH terminus of HERG harbors a large number of LQTS mutations and its removal prevents functional expression for reasons that remain unknown. In this study, we show that the COOH terminus of HERG is required for normal trafficking of the ion channel. We have identified a region critical for trafficking between residues 860 and 899 that includes a novel missense mutation at amino acid 861 (HERGN861I). Truncations or deletion of residues 860-899, characterized in six different expression systems including a cardiac cell line, resulted in decreased expression levels and an absence of the mature glycosylated form of the HERG protein. Deletion of this region did not interfere with the formation of tetramers but caused retention of the assembled ion channels within the endoplasmic reticulum. Consequently, removal of residues 860-899 resulted in the absence of the ion channels from the cell surface and a more rapid turnover rate than the wild type channels, which was evident very early in biogenesis. This study reveals a novel role of the COOH terminus in the normal biogenesis of HERG channels and suggests defective trafficking as a common mechanism for abnormal channel function resulting from mutations of critical COOH-terminal residues, including the LQTS mutant HERGN861I. PMID- 12885766 TI - The coactivator p/CIP/SRC-3 facilitates retinoic acid receptor signaling via recruitment of GCN5. AB - p/CIP/SRC-3 is a member of a family of steroid receptor coactivators/nuclear receptor coactivators (SRC/NCoA) proteins that mediate the transcriptional effects of nuclear hormone receptors (NRs). Using deletion analysis we have mapped the location of two distinct activation domains in p/CIP (AD1 and AD2) capable of activating transcription in mammalian cells when fused to the Gal4-DNA binding domain. In addition to AD1 being coincident with the interaction domain for CBP, we demonstrate a novel in vivo interaction between the AD1 and GCN5. Overexpression of a Gal4-AD1 fusion protein in yeast leads to growth arrest that is relieved by mutation of genes encoding components of the SAGA complex including GCN5, ADA3, and SPT7. In addition, the AD1 of p/CIP and the ADA3 gene are shown to be essential for retinoic acid receptor alpha-dependent transcription in yeast. Transient transfection assays in mammalian cells indicate that GCN5 cooperates with p/CIP as a coactivator of RAR alpha-dependent transcription. Down-regulation of GCN5 using small interfering RNA in mammalian cells indicates that the AD1 domain and the RAR beta promoter activity are dependent, in part, on GCN5. Mutational analysis of AD1 has identified two helical motifs that are required for interactions with GCN5 and CBP. Taken together, these results support a model by which p/CIP functions as a ligand dependent adapter, through specific protein-protein interactions with AD1, to recruit members from at least two distinct families of acetyltransferase proteins to NRs. PMID- 12885767 TI - Engineering the phosphoinositide-binding profile of a class I pleckstrin homology domain. AB - Pleckstrin homology (PH) domains are protein modules that bind with varying degrees of affinity and specificity membrane phosphoinositides. Previously we have shown that although the PH domains of the Ras GTPase-activating proteins GAP1m and GAP1IP4BP are 63% identical at the amino acid level they possess distinct phosphoinositide-binding profiles. The GAP1m PH domain binds phosphatidylinositol 3,4,5-trisphosphate (PtdIns(3,4,5)P3), whereas the domain from GAP1IP4BP binds PtdIns(3,4,5)P3 and phosphatidylinositol 4,5-bisphosphate (PtdIns(4,5)P2) equally well. These phosphoinositide specificities are translated into distinct subcellular localizations. GAP1m is cytosolic and undergoes a rapid PtdIns(3,4,5)P3-dependent association with the plasma membrane following growth factor stimulation. In contrast, GAP1IP4BP is constitutively associated, in a PtdIns(4,5)P2-dependent manner, with the plasma membrane (Cozier, G. E., Lockyer, P. J., Reynolds, J. S., Kupzig, S., Bottomley, J. R., Millard, T., Banting, G., and Cullen, P. J. (2000) J. Biol. Chem. 275, 28261-28268). In the present study, we have used molecular modeling to identify residues in the GAP1IP4BP PH domain predicted to be required for high affinity binding to PtdIns(4,5)P2. This has allowed the isolation of a mutant, GAP1IP4BP-(K591T), which while retaining high affinity for PtdIns(3,4,5)P3 has a 6-fold reduction in its affinity for PtdIns(4,5)P2. Importantly, GAP1IP4BP-(K591T) is predominantly localized to the cytosol and undergoes a PtdIns(3,4,5)P3-dependent association with the plasma membrane following growth factor stimulation. We have therefore engineered the phosphoinositide-binding profile of the GAP1IP4BP PH domain, thereby emphasizing that subtle changes in PH domain structure can have a pronounced effect on phosphoinositide binding and the subcellular localization of GAP1IP4BP. PMID- 12885768 TI - Topogenesis of two transmembrane type K+ channels, Kir 2.1 and KcsA. AB - Potassium channels, which control the passage of K+ across cell membranes, have two transmembrane segments, M1 and M2, separated by a hydrophobic P region containing a highly conserved signature sequence. Here we analyzed the membrane topogenesis characteristics of the M1, M2, and P regions in two animal and bacterial two-transmembrane segment-type K+ channels, Kir 2.1 and KcsA, using an in vitro translation and translocation system. In contrast to the equivalent transmembrane segment, S5, in the voltage-dependent K+ channel, KAT1, the M1 segment in KcsA, was found to have a strong type II signal-anchor function, which favors the Ncyt/Cexo topology. The N-terminal cytoplasmic region was required for efficient, correctly orientated integration of M1 in Kir 2.1. Analysis of N terminal modification by in vitro metabolic labeling showed that the N terminus in Kir 2.1 was acetylated. The hydrophobic P region showed no topogenic function, allowing it to form a loop, but not a transmembrane structure in the membrane; this region was transiently exposed in the endoplasmic reticulum lumen during the membrane integration process. M2 was found to possess a stop-transfer function and a type I signal-anchor function, enabling it to span the membrane. The C terminal cytoplasmic region in KcsA was found to affect the efficiency with which the M2 achieved their final structure. Comparative topogenesis studies of Kir 2.1 and KcsA allowed quantification of the relative contributions of each segment and the cytoplasmic regions to the membrane topology of these two proteins. The membrane topogenesis of the pore-forming structure is discussed using results for Kir 2.1, KcsA, and KAT1. PMID- 12885769 TI - Presenilins mutated at Asp-257 or Asp-385 restore Pen-2 expression and Nicastrin glycosylation but remain catalytically inactive in the absence of wild type Presenilin. AB - The Presenilins are part of the gamma-secretase complex that is involved in the regulated intramembrane proteolysis of amyloid precursor protein and other type I integral membrane proteins. Nicastrin, Pen-2, and Aph1 are the other proteins of this complex. The Presenilins probably contribute the catalytic activity to the protease complex. However, several investigators reported normal Abeta-peptide generation in cells expressing Presenilins mutated at the putative catalytic site residue Asp-257, contradicting this hypothesis. Because endogenously expressed wild type Presenilin could contribute to residual gamma-secretase activity in these experiments, we have reinvestigated the problem by expressing mutated Presenilins in a Presenilin-negative cell line. We confirm that Presenilins with mutated Asp residues are catalytically inactive. Unexpectedly, these mutated Presenilins are still partially processed into amino- and carboxyl-terminal fragments by a "Presenilinase"-like activity. They are also able to rescue Pen-2 expression and Nicastrin glycosylation in Presenilin-negative cells and become incorporated into large approximately 440-kDa complexes as assessed by blue native gel electrophoresis. Our study demonstrates that the catalytic activity of Presenilin and its other functions in the generation, stabilization, and transport of the gamma-secretase complex can be separated and extends the concept that Presenilins are multifunctional proteins. PMID- 12885770 TI - The RING finger protein RNF4, a co-regulator of transcription, interacts with the TRPS1 transcription factor. AB - The TRPS1 gene encodes a repressor of GATA-mediated transcription. Mutations in this gene cause the tricho-rhino-phalangeal syndromes, but the affected pathways are unknown. In a yeast two-hybrid screen with the C-terminal part of the murine Trps1 protein as bait, we obtained three yeast clones encoding two overlapping fragments of the 194 amino acids RING finger protein 4 (Rnf4). The overlap narrows down the Trps1-binding region within Rnf4 to amino acids 6-65. This region in Rnf4 is also known to interact with several proteins including steroid receptors. By using truncated Trps1 constructs, the Rnf4-binding region in Trps1 could be assigned to amino acids 985-1184 of 1281. This 200 amino acid region of Trps1 does not contain any predicted protein-protein interacting motif. Complex formation between the human proteins TRPS1 and RNF4 was verified by co immunoprecipitation from transfected and native mammalian cells. Confocal laser scanning microscopy revealed that the endogenous proteins are located in distinct structures of the nucleus. Using a luciferase reporter assay, we could demonstrate that the repressional function of TRPS1 is inhibited by RNF4. This finding suggests that RNF4 is a negative regulator of TRPS1 activity. PMID- 12885771 TI - beta-Catenin is required for specification of proximal/distal cell fate during lung morphogenesis. AB - The lungs are divided, both structurally and functionally, into two distinct components, the proximal airways, which conduct air, and the peripheral airways, which mediate gas exchange. The mechanisms that control the specification of these two structures during lung development are currently unknown. Here we show that beta-catenin signaling is required for the formation of the distal, but not the proximal, airways. When the gene for beta-catenin was conditionally excised in epithelial cells of the developing mouse lung prior to embryonic day 14.5, the proximal lung tubules grew and differentiated appropriately. The mice, however, died at birth because of respiratory failure. Analysis of the lungs by in situ hybridization and immunohistochemistry, using molecular markers of the epithelial and mesenchymal components of both proximal and peripheral airways, showed that the lungs were composed primarily of proximal airways. These observations establish, for the first time, both the sites and timing of specification of the proximal and peripheral airways in the developing lung, and that beta-catenin is one of the essential components of this specification. PMID- 12885772 TI - CALEB/NGC interacts with the Golgi-associated protein PIST. AB - CALEB/NGC is a neural member of the epidermal growth factor protein family expressed in axon and synapse-rich areas of the nervous system and shown to be important for neurite formation. It can bind to the extracellular matrix proteins tenascin-R and tenascin-C. Here we show that CALEB/NGC interacts with the Golgi associated protein PIST. PIST was originally described as an interaction partner of the small GTPase TC10 and was then found to be Golgi-associated by binding to syntaxin-6 and to be important for the transport of frizzled proteins and the cystic fibrosis transmembrane conductance regulator to the plasma membrane. In addition, PIST was demonstrated to be involved in autophagy and linked to processes of neurodegeneration. CALEB/NGC interacts with PIST in the yeast two hybrid system. This interaction can be confirmed by co-immunoprecipitations and co-localization studies. The juxtamembrane cytoplasmic peptide segment of CALEB/NGC, highly conserved during evolution, mediates the binding to PIST. CALEB/NGC co-localizes with PIST in the Golgi apparatus of transfected COS7 cells and in Golgi-derived vesicles after brefeldin A or nocodazole treatment. Co localization studies in primary hippocampal cells and analysis of Purkinje cells of colchicine-treated rats, serving as an in vivo model system to block microtubule-dependent transport processes, support the view that PIST is an interaction partner of CALEB/NGC and implicate that this interaction may play a role in the intracellular transport of CALEB/NGC. PMID- 12885773 TI - Skeletal muscle signaling pathway through the dystrophin glycoprotein complex and Rac1. AB - The dystrophin glycoprotein complex has been proposed to be involved in signal transduction. Here we have shown that laminin binding causes syntrophin to recruit Rac1 from the rabbit skeletal muscle. Laminin-Sepharose and syntrophin Sepharose bind a protein complex containing Rac1 from the muscle membranes. The presence of heparin, which inhibits laminin interactions, prevents recruitment of Rac1. The dystrophin glycoprotein complex recruits Rac1 via syntrophin through a Grb2.Sos1 complex. A syntrophin antibody also prevents recruitment of Rac1, suggesting that the signaling complex requires syntrophin. PAK1 is in turn bound by Rac1. c-Jun NH2-terminal kinase-p46 is phosphorylated and activated only when laminin is present, and the p54 isoform is activated when laminin is depleted or binding is inhibited with heparin. In the presence of laminin, c-Jun is activated in both skeletal muscle microsomes and in C2C12 myoblasts, and proliferation increases in C2C12 myoblasts. We postulate that this pathway signals muscle homeostasis and hypertrophy. PMID- 12885774 TI - Identification of human intestinal alkaline sphingomyelinase as a novel ecto enzyme related to the nucleotide phosphodiesterase family. AB - Alkaline sphingomyelinase (alk-SMase) hydrolyzes dietary sphingomyelin and generates sphingolipid messengers in the gut. In the present study, we purified the enzyme, identified a part of the amino acid sequence, and found a cDNA in the GenBank coding for the protein. The cDNA contains 1841 bp, and the open reading frame encodes 458 amino acids. Transient expression of the cDNA linked to a Myc tag in COS-7 cells increased alk-SMase activity in the cell extract by 689-fold and in the medium by 27-fold. High activity was also identified in the anti-Myc immunoprecipitated proteins and the proteins cross-reacted with anti-human alk SMase. Northern blotting of human intestinal tissues found high levels of alk SMase mRNA in the intestine and liver. The amino acid sequence shared no similarity with acid and neutral SMases but was related to the ecto-nucleotide phosphodiesterase (NPP) family with 30-36% identity to human NPPs. Alk-SMase has a predicted signal peptide domain at the N terminus and a signal anchor domain at the C terminus. The ion-binding sites and the catalytic residue of NPPs were conserved, but the substrate specificity domain was modified. Alk-SMase had no detectable nucleotidase activity, but its activity against sphingomyelin could be inhibited by orthovanadate, imidazole, and ATP. In contrast to NPPs, alk-SMase activity was not stimulated by divalent metal ions but inhibited by Zn2+. Differing from NPP2, the alk-SMase cleaved phosphocholine but not choline from lysophosphatidylcholine. Phylogenetic tree indicated that the enzyme is a new branch derived from the NPP family. Two cDNA sequences of mouse and rat that shared 83% identity to human alk-SMase were identified in the GenBank. In conclusion, we identified the amino acid and cDNA sequences of human intestinal alk-SMase, and found that it is a novel ecto-enzyme related to the NPP family with specific features essential for its SMase activity. PMID- 12885775 TI - The association of alpha-synuclein with membranes affects bilayer structure, stability, and fibril formation. AB - The aggregation of alpha-synuclein is believed to be a critical factor in the etiology of Parkinson's disease. alpha-Synuclein is an abundant neuronal protein of unknown function, which is enriched in the presynaptic terminals of neurons. Although alpha-synuclein is found predominantly in the cytosolic fractions, membrane-bound alpha-synuclein has been suggested to play an important role in fibril formation. The effects of alpha-synuclein on lipid bilayers of different compositions were determined using fluorescent environment-specific probes located at various depths. alpha-Synuclein-membrane interactions were found to affect both protein and membrane properties. Our results indicate that in addition to electrostatic interactions, hydrophobic interactions are important in the association of the protein with the bilayer, and lead to disruption of the membrane. The latter was observed by atomic force microscopy and fluorescent dye leakage from vesicles. The kinetics of alpha-synuclein fibril formation were significantly affected by the protein association and subsequent membrane disruption, and reflected the conformation of alpha-synuclein. The ability of alpha-synuclein to disrupt membranes correlated with the binding affinity of alpha-synuclein for the particular membrane composition, and to the induced helical conformation of alpha-synuclein. Protofibrillar or fibrillar alpha synuclein caused a much more rapid destruction of the membrane than soluble monomeric alpha-synuclein, indicating that protofibrils (oligomers) or fibrils are likely to be significantly neurotoxic. PMID- 12885776 TI - The energetics of Pex5p-mediated peroxisomal protein import. AB - Most newly synthesized peroxisomal matrix proteins are targeted to the organelle by Pex5p, the peroxisomal cycling receptor. According to current models of peroxisomal biogenesis, Pex5p interacts with cargo proteins in the cytosol and transports them to the peroxisomal membrane. After delivering the passenger protein into the peroxisomal matrix, Pex5p returns to the cytosol to catalyze additional rounds of transportation. Obviously, such cyclic pathway must require energy, and indeed, data confirming this need are already available. However, the exact step(s) of this cycle where energy input is necessary remains unclear. Here, we present data suggesting that insertion of Pex5p into the peroxisomal membrane does not require ATP hydrolysis. This observation raises the possibility that at the peroxisomal membrane ATP is needed predominantly (if not exclusively) downstream of the protein translocation step to reset the Pex5p-mediated transport system. PMID- 12885777 TI - Regulation of MuSK expression by a novel signaling pathway. AB - MuSK is a receptor tyrosine kinase essential for neuromuscular junction formation. Expression of the MuSK gene is tightly regulated during development and at the neuromuscular junction. However, little is known about molecular mechanisms regulating its gene expression. Here we report a characterization of the promoter of the mouse MuSK gene. The transcription of MuSK starts at multiple sites with a major site 51 nt upstream of the translation start site. We have identified an E-box-like cis-element that is both required and sufficient for differentiation-dependent transcription. Interestingly, the promoter activity of the MuSK gene did not respond to neuregulin, a factor believed to mediate the synapse-specific transcription of acetylcholine receptor subunit genes. Rather, MuSK expression is increased in muscle cells stimulated with Wnt or at conditions when the Wnt signaling was activated. These results suggest a novel mechanism for the MuSK synapse-specific expression. PMID- 12885778 TI - Viral stress-inducible protein p56 inhibits translation by blocking the interaction of eIF3 with the ternary complex eIF2.GTP.Met-tRNAi. AB - Viral stress-inducible protein p56 is produced in response to viral stress inducing agents such as double-stranded RNA and interferon, as well as other poorly understood mechanisms of viral infection. It has been shown previously that p56 is able to bind the eukaryotic initiation factor 3e(eIF3e) (p48/Int-6) subunit of the eukaryotic translation initiation factor eIF3 and function as an inhibitor of translation in vitro and in vivo. The exact mechanism by which p56 is able to interfere with protein synthesis is not understood. Based on the known roles of eIF3 in the initiation pathway, we employed assays designed to individually look at specific functions of eIF3 and the effect of p56 on these eIF3-mediated functions. These assays examined the effect of p56 on ribosome dissociation, the eIF3.eIF4F interaction, and enhancement of the ternary complex eIF2.GTP.Met-tRNAi formation. Here we report that p56 is able to inhibit translation initiation specifically at the level of eIF3.ternary complex formation. The effect of p56-mediated inhibition was also examined in two different contexts, cap-mediated and encephalomyocarditis virus internal ribosomal entry site-mediated translation. Whereas cap-dependent initiation was severely inhibited by p56, internal ribosomal entry site-mediated translation appeared to be insensitive to p56. PMID- 12885779 TI - The water-water cycle is essential for chloroplast protection in the absence of stress. AB - Maintaining electron flow through the photosynthetic apparatus, even in the absence of a sufficient amount of NADP+ as an electron acceptor, is essential for chloroplast protection from photooxidative stress. At least two different pathways are thought to participate in this process, i.e. cyclic electron flow and the water-water cycle. Although the function of the water-water cycle was inferred from a number of biochemical and physiological studies, genetic evidence for the function of this cycle is very limited. Here we show that knockdown Arabidopsis plants with suppressed expression of the key water-water cycle enzyme, thylakoid-attached copper/zinc superoxide dismutase (KD-SOD), are suppressed in their growth and development. Chloroplast size, chlorophyll content, and photosynthetic activity were also reduced in KD-SOD plants. Microarray analysis of KD-SOD plants, grown under controlled conditions, revealed changes in transcript expression consistent with an acclimation response to light stress. Although a number of transcripts involved in the defense of plants from oxidative stress were induced in KD-SOD plants, and seedlings of KD-SOD plants were more tolerant to oxidative stress, these mechanisms were unable to compensate for the suppression of the water-water cycle in mature leaves. Thus, the localization of copper/zinc superoxide dismutase at the vicinity of photosystem I may be essential for its function. Our studies provide genetic evidence for the importance of the water-water cycle in protecting the photosynthetic apparatus of higher plants from photooxidative damage. PMID- 12885780 TI - Mechanism of group IVA cytosolic phospholipase A(2) activation by phosphorylation. AB - Group IVA cytosolic phospholipase A2 (cPLA2) has been shown to play a critical role in the agonist-induced release of arachidonic acid. To understand the mechanism by which phosphorylation of Ser505 and Ser727 activates cPLA2, we systematically analyzed the effects of S505A, S505E, S727A, S727E, S505A/S727A, S505A/S727E, and S505E/S727E mutations on its enzyme activity and membrane affinity. In vitro membrane binding measurements showed that S505A has lower affinity than the wild type or S505E for phosphatidylcholine membranes, which is exclusively due to faster desorption of the membrane-bound S505A. In contrast, neither S727A nor S727E mutation had a significant effect on the phosphatidylcholine vesicle binding affinity of cPLA2. The difference in in vitro membrane affinity between wild type (or S505E) and S505A increased with the decrease in Ca2+ concentration, reaching >60-fold at 2.5 microm Ca2+. When HEK293 cells transfected with cPLA2 and mutants were stimulated with ionomycin, the wild type and S505E translocated to the perinuclear region and caused the arachidonic acid release at 0.4 microm Ca2+, whereas S505A showed no membrane translocation and little activity to release arachidonic acid. Further mutational analysis of hydrophobic residues in the active site rim (Ile399, Leu400, and Leu552) indicate that a main role of the Ser505 phosphorylation is to promote membrane penetration of these residues, presumably by inducing a conformational change of the protein. These enhanced hydrophobic interactions allow the sustained membrane interaction of cPLA2 in response to transient calcium increases. On the basis of these results, we propose a mechanism for cPLA2 activation by calcium and phosphorylation. PMID- 12885781 TI - Molecular pathway for cancer metastasis to bone. AB - The molecular mechanism leading to the cancer metastasis to bone is poorly understood but yet determines prognosis and therapy. Here, we define a new molecular pathway that may account for the extraordinarily high osteotropism of prostate cancer. By using SPARC (secreted protein, acidic and rich in cysteine) deficient mice and recombinant SPARC, we demonstrated that SPARC selectively supports the migration of highly metastatic relative to less metastatic prostate cancer cell lines to bone. Increased migration to SPARC can be traced to the activation of integrins alphaVbeta3 and alphaVbeta5 on tumor cells. Such activation is induced by an autocrine vascular endothelial growth factor (VEGF)/VEGF receptor (VEGFR)-2 loop on the tumor cells, which also supports the growth and proliferation of prostate cancer cells. A consequence of SPARC recognition by alphaVbeta5 is enhanced VEGF production. Thus, prostate cancer cells expressing VEGF/VEGFR-2 will activate alphaVbeta3 and alphaVbeta5 on their surface and use these integrins to migrate toward SPARC in bone. Within the bone environment, SPARC engagement of these integrins will stimulate growth of the tumor and further production of VEGF to support neoangiogenesis, thereby favoring the development of the metastatic tumor. Supporting this model, activated integrins were found to colocalize with VEGFR-2 in tissue samples of metastatic prostate tumors from patients. PMID- 12885782 TI - Interaction of Bordetella pertussis adenylate cyclase with CD11b/CD18: Role of toxin acylation and identification of the main integrin interaction domain. AB - Adenylate cyclase toxin (CyaA) is one of the major virulence factors produced by Bordetella pertussis, the whooping cough agent. CyaA belongs to the repeat in toxin protein family and requires a post-translational fatty acylation to form cation-selective channels in target cell membranes and to penetrate into cytosol. We have demonstrated recently that CyaA uses the alphaMbeta2 integrin (CD11b/CD18) as a specific cellular receptor. Here we show that the acylation of CyaA is required for a productive and tight interaction of the toxin with cells expressing CD11b. In addition, we demonstrate that the catalytic domain is not required for binding of CyaA to CD11b and that the main integrin interacting domain of CyaA is located in its glycine/aspartate-rich repeat region. These data decipher, for the first time, the interaction of CyaA with CD11b-positive cells and open new prospects for understanding the interaction of Bordetella pertussis with innate and adaptive immune systems. PMID- 12885783 TI - Disassembly and degradation of photosystem I in an in vitro system are multievent, metal-dependent processes. AB - An in vitro system was created to study the process of membrane protein degradation by using photosystem I (PS1) as a model membrane protein. Purified chloroplast membranes were incubated at 30 degrees C in a defined buffer along with various extracts or reagents to reconstitute the disassembly and degradation of PS1, which was monitored by a variety of techniques that probe the integrity of the PS1 complex: photo-biochemical assays, semi-native gel electrophoresis, low temperature fluorescence spectroscopy, and immunoblots using antibodies against different PS1 subunits. During a typical time course, degradation of PS1 appeared to be a multievent process, with disassembly of the complex preceding proteolysis of the subunits. The first change seen was a rapid (<5 min) decrease in PS1 photochemical activity. This was followed by a diminution of far-red fluorescence emission from the core antenna of PS1 and a slower disassembly of the PS1 chlorophyll-protein core complex, as visualized by semi-native gel electrophoresis. Surprisingly, the latter was not accompanied by a similar rate of proteolysis of the PsaA core subunit. In contrast, addition of soluble proteases caused rapid loss of immuno-detectable PS1 polypeptides and cleavage of the major PS1 polypeptides in interhelical loops. The in vitro degradation process was time- and temperature-dependent but did not require ATP, GTP, or soluble chloroplast proteins. Chelation of divalent cations by EDTA inhibited the later steps of disassembly and proteolysis, and this effect could be reversed by addition of micromolar Zn2+, with Co2+ and Ca2+ providing somewhat lower activity. PMID- 12885784 TI - Asymmetric cross-inhibition between GABAA and glycine receptors in rat spinal dorsal horn neurons. AB - Presynaptic nerve terminals of inhibitory synapses in the dorsal horn of the spinal cord and brain stem can release both GABA and glycine, leading to coactivation of postsynaptic GABAA and glycine receptors. In the present study we have analyzed functional interactions between GABAA and glycine receptors in acutely dissociated neurons from rat sacral dorsal commissural nucleus. Although the application of GABA and glycine activates pharmacologically distinct receptors, the current induced by a simultaneous application of these two transmitters was less than the sum of currents induced by applying two transmitters separately. Sequential application of glycine and GABA revealed that the GABA-evoked current is more affected by glycine than glycine-evoked responses by GABA. Activation of glycine receptors decreased the amplitude and accelerated the rate of desensitization of GABA-induced currents. This asymmetric cross inhibition is reversible, dependent on the agonist concentration applied, but independent of both membrane potential and intracellular calcium concentration or changes in the chloride equilibrium potential. During sequential applications, the asymmetric cross-inhibition was prevented by selective GABAA or glycine receptor antagonists, suggesting that occupation of binding sites did not suffice to induce glycine and GABAA receptors functional interaction, and receptor channel activation is required. Furthermore, inhibition of phosphatase 2B, but not phosphatase 1 or 2A, prevented GABAA receptor inhibition by glycine receptor activation, whereas inhibition of phosphorylation pathways rendered cross-talk irreversible. Taken together, our results demonstrated that there is an asymmetric cross-inhibition between glycine and GABAA receptors and that a selective modulation of the state of phosphorylation of GABAA receptor and/or mediator proteins underlies the asymmetry in the cross-inhibition. PMID- 12885785 TI - The role of HIF-1 alpha in transcriptional regulation of the proximal tubular epithelial cell response to hypoxia. AB - Epithelial cells of the kidney represent a primary target for hypoxic injury in ischemic acute renal failure (ARF); however, the underlying transcriptional mechanism(s) remain undefined. In this study, human proximal tubular epithelial cells (HK-2) exposed to hypoxia in vitro demonstrated a non-lethal but dysfunctional phenotype, closely reflective of the epithelial pathobiology of ARF. HK-2 cells exposed to hypoxia demonstrated increased paracellular permeability, decreased proliferation, loss of tight junctional integrity, and significant actin disassembly in the absence of cell death. Microarray analysis of transcriptomic changes underlying this response identified a distinct cohort of 48 genes with a closely shared hypoxia-dependent expression profile. Within this hypoxia-sensitive cluster were genes identified previously as hypoxia inducible factor-1 (HIF-1)-dependent (e.g. vascular endothelial growth factor and adrenomedullin) as well as genes not previously known to be hypoxia-responsive (e.g. stanniocalcin 2). In hypoxia, HIF-1 bound to evolutionarily conserved hypoxia-response elements (HRE) in the promoters of these genes as well as to the HRE consensus motif. A further subset of these genes, not associated with transcriptional regulation by HIF-1, was also present, suggesting alternative HIF 1-independent pathways. Overexpression of HIF-1 alpha in normoxia induced the expression of a significant number of the hypoxia-dependent genes; however, it did not induce the pathophysiologic epithelial response. In summary, hypoxia elicited alterations in renal proximal tubular epithelial cells in vitro closely resemble the epithelial pathophysiology of ARF. Our data indicate that although this event may rely heavily on HIF-1-dependent gene transcription, it is likely that separate hypoxia-dependent transcriptional regulators also play a role. PMID- 12885786 TI - LIS1 missense mutations: variable phenotypes result from unpredictable alterations in biochemical and cellular properties. AB - Mutations in one allele of the human LIS1 gene cause a severe brain malformation, lissencephaly. Although most LIS1 mutations involve deletions, several point mutations with a single amino acid alteration were described. Patients carrying these mutations reveal variable phenotypic manifestations. We have analyzed the functional importance of these point mutations by examining protein stability, folding, intracellular localization, and protein-protein interactions. Our data suggest that the mutated proteins were affected at different levels, and no single assay could be used to predict the lissencephaly phenotype. Most interesting are those mutant proteins that retain partial folding and interactions. In the case of LIS1 mutated in F31S, the cellular phenotype may be modified by overexpression of specific interacting proteins. Overexpression of the PAF-AH alpha1 subunit dissolved aggregates induced by this mutant protein and increased its half-life. Overexpression of NudE or NudEL localized this mutant protein to spindle poles and kinetochores but had no effect on protein stability. Our results implicate that there are probably different biochemical and cellular mechanisms obstructed in each patient yielding the varied lissencephaly phenotypes. PMID- 12885787 TI - Inhibition of the norepinephrine transporter by the venom peptide chi-MrIA. Site of action, Na+ dependence, and structure-activity relationship. AB - chi-Conopeptide MrIA (chi-MrIA) is a 13-residue peptide contained in the venom of the predatory marine snail Conus marmoreus that has been found to inhibit the norepinephrine transporter (NET). We investigated whether chi-MrIA targeted the other members of the monoamine transporter family and found no effect of the peptide (100 microM) on the activity of the dopamine transporter and the serotonin transporter, indicating a high specificity of action. The binding of the NET inhibitors, [3H]nisoxetine and [3H]mazindol, to the expressed rat and human NET was inhibited by chi-MrIA with the conopeptide displaying a slight preference toward the rat isoform. For both radioligands, saturation binding studies showed that the inhibition by chi-MrIA was competitive in nature. It has previously been demonstrated that chi-MrIA does not compete with norepinephrine, unlike classically described NET inhibitors such as nisoxetine and mazindol that do. This pattern of behavior implies that the binding site for chi-MrIA on the NET overlaps the antidepressant binding site and is wholly distinct from the substrate binding site. The inhibitory effect of chi-MrIA was found to be dependent on Na+ with the conopeptide becoming a less effective blocker of [3H]norepinephrine by the NET under the conditions of reduced extracellular Na+. In this respect, chi-MrIA is similar to the antidepressant inhibitors of the NET. The structure-activity relationship of chi-MrIA was investigated by alanine scanning. Four residues in the first cysteine-bracketed loop of chi-MrIA and a His in loop 2 played a dominant role in the interaction between chi-MrIA and the NET. H alpha chemical shift comparisons indicated that side-chain interactions at these key positions were structurally perturbed by the replacement of Gly-6. From these data, we present a model of the structure of chi-MrIA that shows the relative orientation of the key binding residues. This model provides a new molecular caliper for probing the structure of the NET. PMID- 12885788 TI - Blunted fenfluramine-evoked prolactin secretion in hypertensive rats. AB - Plasma prolactin (PRL) levels after acute administration of fenfluramine (FEN) have been used as a probe of brain serotonin activity. FEN-evoked increases in PRL levels inversely correlate with arterial blood pressure (ABP) in humans (Muldoon et al. Hypertension. 1998;32:972-975), thereby suggesting that brain serotonin activity may be reduced in hypertension. The present study sought to determine whether the relation between FEN-evoked PRL levels and ABP was present in two rat models of hypertension. Experiments were performed in awake male rats that were instrumented with femoral arterial and venous catheters 2 days before experiments. FEN (3.0 mg/kg IV) significantly increased plasma PRL levels in both spontaneously hypertensive rats (SHR) and Wistar-Kyoto rats (WKY); however, FEN evoked PRL levels were significantly lower in SHR compared with WKY, though baseline levels were similar between strains. Similar results were obtained in rats with chronic hypertension produced by figure-8 renal wrap plus contralateral nephrectomy. In contrast, the increase in PRL levels evoked by the serotonin receptor agonist m-CPP or the dopamine receptor antagonist eticlopride did not differ between SHR and WKY, indicating that PRL secretion is not generally blunted in chronic hypertensive rats. Furthermore, FEN-evoked PRL levels were not attenuated in rats made acutely hypertensive by an infusion of the alpha adrenergic agonist phenylephrine. Thus, the present findings are consistent with the human data and suggest that chronic hypertension is associated with a presynaptic alteration in brain serotonin function. PMID- 12885789 TI - Na+/Ca2+ exchanger: target for oxidative stress in salt-sensitive hypertension. AB - The Na+/Ca2+ exchanger regulates intracellular calcium ([Ca2+]i), and attenuation of Na+/Ca2+ exchange by oxidative stress might lead to dysregulation of [Ca2+]i. We have shown that the Na+/Ca2+ exchanger differs functionally and at the amino acid level between salt-sensitive and salt-resistant rats. Therefore, the purpose of these studies was to determine how oxidative stress affects the activities of the 2 Na+/Ca2+ exchangers that we cloned from mesangial cells of salt-resistant (RNCX) and salt-sensitive (SNCX) Dahl/Rapp rats. The effects of oxidative stress on exchanger activity were examined in cells expressing RNCX or SNCX by assessing 45Ca2+ uptake (reverse mode) and [Ca2+]i elevation (forward mode) in the presence and absence of H2O2 and peroxynitrite. Our results showed that 45Ca2+ uptake in SNCX cells was attenuated at 500 and 750 micromol/L H2O2 (63+/-12% and 25+/-7%, respectively; n=16) and at 50 and 100 micromol/L peroxynitrite (47+/-9% and 22+/ 9%, respectively; n=16). In RNCX cells, 45Ca2+ uptake was attenuated at only 750 and 100 micromol/L H2O2 and peroxynitrite (61+/-9% and 63+/-6%, respectively; n=16). In addition, the elevation in [Ca2+]i was greater in SNCX cells than in RNCX cells in response to 750 micromol/L H2O2 (58+/-5.5 vs 17+/-4.1 nmol/L; n=13) and 100 micromol/L peroxynitrite (33+/-5 vs 11+/-6 nmol/L; n=19). The enhanced impairment of SNCX activity by oxidative stress might contribute to the dysregulation of [Ca2+]i that is found in this model of salt-sensitive hypertension. PMID- 12885790 TI - Daily life blood pressure changes are steeper in hypertensive than in normotensive subjects. AB - Target organ damage in hypertensive patients is related to their increased average blood pressure and greater 24-hour blood pressure variability. Whether the rate of blood pressure changes is also greater in hypertension, producing a greater stress on arterial walls, is not known, however. Our study aimed at addressing this issue by computer analysis of 24-hour ambulatory intra-arterial blood pressure recordings in 34 subjects (29 males), 13 normotensive subjects and 21 uncomplicated hypertensive subjects (mean age+/-SD, 40.4+/-11.8 years). The number, slope (mm Hg/s), and length (beats) of systolic blood pressure ramps of 3 or more consecutive beats characterized by a progressive increase (+) or reduction (-) in systolic blood pressure of at least 1 mm Hg per beat were computed for each hour and for the whole 24-hour period. Twenty-four-hour average systolic blood pressure was 112.9+/-2.1 and 159.4+/-5.7 mm Hg in normotensive and hypertensive subjects, respectively. Over the 24 hours, the number and length of systolic blood pressure ramps were similar in both groups, whereas the slope was markedly different (24-hour mean+/-SE slope, 4.80+/-0.30 in normotensives and 6.50+/-0.40 mm Hg/s in hypertensives, P<0.05). Ramp slope was not influenced by age or reflex pulse interval changes, but it was greater for higher ramp initial systolic blood pressure values. Thus, in daily life, hypertensive subjects are characterized by steeper blood pressure changes than normotensives, and this, regardless of the mechanisms, may have clinical implications, because it may be associated with greater traumatic effect on the vessel walls of hypertensive patients. PMID- 12885791 TI - Urotensin II acts centrally to increase epinephrine and ACTH release and cause potent inotropic and chronotropic actions. AB - Urotensin II is a small peptide whose receptor was recently identified in mammals as the orphan G protein-coupled receptor-14. The reported cardiovascular responses to systemic urotensin II administration are variable, and there is little detailed information on its central cardiovascular actions. We examined the cardiovascular and humoral actions of intracerebroventricular urotensin II (0.02 and 0.2 nmol/kg and vehicle) and intravenous urotensin II (2, 20, and 40 nmol/kg and vehicle) in conscious ewes previously surgically implanted with flow probes and intracerebroventricular guide tubes. Two hours after intracerebroventricular infusion of urotensin II (0.2 nmol/kg over 1 hour; n=5), heart rate (+56+/-13 beats per minute [bpm]), dF/dt (an index of cardiac contractility; +533+/-128 L x min(-1) x s(-1)), and cardiac output (+3.4+/-0.4 L/min) increased significantly compared with vehicle, as did renal, mesenteric, and iliac blood flows and conductances. Plasma epinephrine, adrenocorticotropic hormone, and glucose levels also increased dramatically (+753+/-166 pg/mL, +14.3+/-3.5 pmol/L, and +7.0+/-1.4 mmol/L, respectively). All of these variables remained elevated for up to 4 hours after infusion. In contrast, 1 hour after intravenous urotensin II (40 nmol/kg bolus; n=6), a sustained tachycardia (+25+/ 8 bpm) ensued, but cardiac output, cardiac contractility, total peripheral conductance, and plasma glucose levels did not change significantly. In summary, this is the first study to show that urotensin II acts centrally to stimulate sympathoadrenal and pituitary-adrenal pathways, resulting in increased adrenocorticotropic hormone and epinephrine release and potent chronotropic and inotropic actions. In contrast, tachycardia was the only major response to intravenous urotensin II. These findings suggest that urotensin II is a novel stimulator of central pathways that mediate responses to alerting stimuli or stress. PMID- 12885792 TI - NADPH oxidase-derived superoxide augments endothelin-1-induced venoconstriction in mineralocorticoid hypertension. AB - Deoxycorticosterone acetate (DOCA)-salt hypertension is characterized by low renin/angiotensin but increased arterial superoxide levels. We have recently reported that the arterial endothelin-1 (ET-1) level is increased, resulting in NADPH oxidase activation and superoxide generation. However, the effect of ET-1 on venous superoxide production and its relation to venoconstriction are unknown. The present study tested the hypotheses that ET-1 stimulates venous NADPH oxidase and superoxide via its ET(A) receptors, resulting in enhanced venoconstriction in DOCA-salt hypertensive rats. Treatment with ET-1 (0.01 to 1 nmol/L), but not the selective ET(B) receptor agonist sarafotoxin s6c, of vena cavas of normal rats concentration-dependently increased superoxide levels, an effect that was abolished by the selective ET(A) receptor antagonist ABT-627. Although the ET-1 level was not increased in the vena cava and plasma, both venous NADPH oxidase activity and superoxide levels were significantly higher in DOCA-salt compared with sham rats. Moreover, ET-1 treatment (10(-9) mol/L, 10 minutes) of isolated vena cavas further elevated superoxide levels in DOCA-salt rats only but not sham rats, an effect that was abrogated by the superoxide scavenger tempol. Similarly, ET-1-induced contractions of isolated vena cavas of DOCA-salt but not sham rats were significantly inhibited by tempol. The NADPH oxidase inhibitor apocynin significantly reduced superoxide levels in vena cavas of DOCA-salt rats and in ET 1-treated vena cavas of normal rats. Finally, in vivo ET(A) receptor blockade by ABT-627 significantly lowered venous superoxide levels and blood pressure in DOCA salt but not sham rats. These results suggest that superoxide contributes to ET-1 induced venoconstriction through an elevated venous NADPH oxidase activity in mineralocorticoid hypertension. PMID- 12885793 TI - Interaction between ACE and ADD1 gene polymorphisms in the progression of IgA nephropathy in Japanese patients. AB - An interaction effect between the angiotensin-converting enzyme insertion/deletion (ACE I/D) and alpha-adducin (ADD1) Gly460Trp polymorphisms (G460W) on blood pressure regulation has recently been suggested, although its significance in the prognosis of renal function in IgA nephropathy (IgAN) has not been fully investigated. Therefore, we evaluated the clinical manifestations and renal prognosis in 276 Japanese patients with histologically proven IgAN with respect to their ACE I/D and ADD1 G460W polymorphisms. The prognosis of renal function was analyzed by Kaplan-Meier survival curves and multivariate Cox proportional-hazards regression models. Baseline data, including blood pressures, proteinuria, renal function, and incidence of hypertension, were similar for the different genotypes of ACE and ADD1. The individual genotypes taken alone were not associated with the progression of renal dysfunction. However, renal survival of patients with the 460WW polymorphism of ADD1 was significantly worse within the group with the II genotype of ACE (Kaplan-Meier, log rank test; chi2=6.062, P=0.0138) but not for those with other ACE genotypes. In the Cox proportional hazards regression model with adjustment for clinical risk factors, including hypertension, proteinuria, and no administration of an angiotensin-converting enzyme inhibitors or angiotensin II receptor blockers, the 460WW variant of ADD1 was a highly significant and independent risk factor only for patients with the ACE II genotype, with a hazard ratio of 3.65 (P=0.0016), but not for those with other ACE genotypes (hazard ratio=0.65, P=0.2902). These findings suggest an interaction between ACE and ADD1 polymorphisms not only on blood pressure regulation but also on the progression of renal dysfunction in patients with IgAN. PMID- 12885794 TI - Attenuated responses to angiotensin II in follitropin receptor knockout mice, a model of menopause-associated hypertension. AB - Activation of the renin-angiotensin system has been implicated in the development of hypertension in menopausal women. We investigated whether blood pressure is elevated and whether angiotensin II (Ang II)-induced vascular reactivity is increased in follitropin receptor knockout (FORKO) female mice. These mice are estrogen-deficient and have characteristics similar to postmenopausal women. Serum estradiol levels were significantly reduced in FORKO versus wild-type mice (1.4+/-0.2 versus 15+/-3 pg/mL, P<0.01). Blood pressure, measured by telemetry, was significantly increased in FORKO (120+/-2/92+/-2 mm Hg) compared with wild type counterparts (110+/-1/85+/-2 mm Hg, P<0.05). Vascular dose responses to acetylcholine (endothelium-dependent dilation) and sodium nitroprusside (endothelium-independent dilation) were not different. Ang II-induced vasoconstriction was blunted in FORKO compared with wild-type mice (P<0.05). Media-to-lumen ratio was significantly increased in FORKO (6.2+/-0.5%) versus control mice (5.2+/-0.3%), indicating vascular remodeling. Aortic*O2- levels, NADH-inducible.O2- generation, and plasma levels of thiobarbituric acid reactive substances (TBARS), indexes of oxidative stress, were not significantly different between wild-type and FORKO mice. Vascular AT1 receptor content, assessed by immunoblotting, was reduced by 40% in FORKO compared with wild-type mice (P<0.01). This was associated with decreased circulating Ang II levels in FORKO versus control mice. These data indicate that FORKO mice have increased blood pressure, vascular remodeling, and attenuated vascular responses to Ang II. Our findings suggest that vascular Ang II signaling is downregulated in female FORKO mice and that Ang II may not play an important role in blood pressure elevation in this model of menopause-associated hypertension. PMID- 12885795 TI - PPARgamma inhibition of cyclooxygenase-2, PGE2 synthase, and inducible nitric oxide synthase in cardiac myocytes. AB - Peroxisome proliferator-activated receptors (PPARs) are ligand-activated transcription factors belonging to the nuclear receptor superfamily. They regulate lipid metabolism, glucose homeostasis, cell proliferation, and differentiation and modulate inflammatory responses. We examined whether PPARgamma is functional in cultured neonatal ventricular myocytes and studied its role in inflammation. Western blots revealed PPARgamma in myocytes. When myocytes were transfected with a PPAR response element reporter plasmid (PPRE-TK luciferase), the PPARgamma activator 15-deoxy-Delta12,14-prostaglandin J2 (15dPGJ2) increased promoter activity, whereas cotransfection of a dominant negative PPARgamma inhibited it. To determine the role of 15dPGJ2 in expression of proinflammatory genes, we tested its effect on interleukin-1beta induction of cyclooxygenase-2 (COX-2). 15dPGJ2 decreased interleukin-1beta stimulation of COX 2 by 40% and PGE2 production by 73%. We next questioned whether 15dPGJ2 was modulating the expression of inducible prostaglandin E2 synthase (PGES) and found that it completely blocked interleukin-1beta induction of PGES. Use of a second PPARgamma agonist, troglitazone, and the selective PPARgamma antagonist GW9662 demonstrated that the effects seen were PPARgamma-dependent. In addition, we found that 15dPGJ2 blocked interleukin-1beta stimulation of inducible nitric oxide synthase (iNOS). We concluded that 15dPGJ2 may play an anti-inflammatory role in a PPARgamma-dependent manner, decreasing COX-2, PGES, and PGE2 production, as well as iNOS expression. PMID- 12885796 TI - Role of Crk-associated substrate in the regulation of vascular smooth muscle contraction. AB - A pool of actin monomers is induced to polymerize into actin filaments during contractile stimulation of smooth muscle. The inhibition of actin dynamics by actin polymerization inhibitors depresses active force generation in smooth muscle. In this study, we hypothesized that Crk-associated substrate plays a role in the regulation of contraction and actin dynamics in vascular smooth muscle. Antisense or sense oligodeoxynucleotides for Crk-associated substrate were introduced into carotid smooth muscle tissues by chemical loading. The treatment of smooth muscle strips with antisense oligodeoxynucleotides inhibited the expression of Crk-associated substrates; it did not influence the expression of actin, myosin heavy chain, and paxillin. Sense oligodeoxynucleotides did not affect the expression of these proteins in smooth muscle tissues. Force generation in response to stimulation with norepinephrine or KCl was significantly lower in antisense-treated muscle strips than in sense-treated strips or in muscle strips not treated with oligodeoxynucleotides. The downregulation of Crk-associated substrate did not attenuate increases in phosphorylation of the 20-kDa regulatory light chain of myosin in response to stimulation with norepinephrine. The increase in F-actin/G-actin ratio during contractile stimulation was significantly inhibited in antisense-treated smooth muscle strips. Contractile activation of smooth muscle increased the association of profilin with actin monomers; the depletion of Crk-associated substrate inhibited the increases in the profilin-actin complex in response to contractile stimulation. These results suggest that Crk-associated substrate is a necessary molecule of signaling cascades that regulate active force generation in smooth muscle. This molecule may regulate actin dynamics in smooth muscle in response to contractile stimulation. PMID- 12885797 TI - Volume status and blood pressure during long-term hemodialysis: role of ventricular stiffness. AB - The importance of volume status on blood pressure in hemodialysis patients has long been recognized. We hypothesized that the enhanced volume dependency of blood pressure is partly determined by ventricular stiffness at end systole. A total of 115 long-term hemodialysis patients were invited to receive a comprehensive, noninvasive cardiovascular examination. End-systolic elastance was determined by using a novel, noninvasive echo-Doppler technique. The positive ratios of the interdialytic systolic blood pressure change vs weight gain during the subsequent 25 hemodialysis sessions were averaged to obtain the volume sensitivity index (mm Hg/kg). The averaged interdialytic weight gain per fat-free mass was correlated significantly with averaged percent change in systolic blood pressure (r=0.45, P<0.001). The estimated end-systolic elastance at baseline was significantly correlated with subsequently observed volume sensitivity index (volume sensitivity index=[1.17xend-systolic elastance]+6.4; r=0.34, P=0.001). End-systolic elastance was also significantly correlated with various vascular function parameters, including effective arterial elastance (r=0.48, P<0.001), pulse wave velocity (r=0.30, P=0.001), carotid augmentation index (r=0.31, P<0.001), and aortic compliance (r=-0.49, P<0.001). The results suggest that end systolic elastance, a direct measure of left ventricular mechanical properties at end systole, is coupled to arterial mechanical properties and predicts the extent of subsequent interdialytic systolic blood pressure rise relative to weight gain. Therefore, ventricular stiffness at end systole is a determinant of the enhanced volume sensitivity of blood pressure in hemodialysis patients. PMID- 12885798 TI - Hypertension and low-level lead exposure: a scientific issue or a matter of faith? PMID- 12885800 TI - Intraperitoneal radioactive phosphorus (32P) versus observation after negative second-look laparotomy for stage III ovarian carcinoma: a randomized trial of the Gynecologic Oncology Group. AB - PURPOSE: The objectives of this prospective randomized study of consolidation therapy were to evaluate recurrence-free survival (RFS), overall survival (OS), and the morbidity of intraperitoneal (IP) chromic phosphate suspension (32P) therapy in patients with stage III epithelial ovarian carcinoma who have no detectable evidence of disease at the second-look laparotomy (SLL) procedure after primary chemotherapy. PATIENTS AND METHODS: In a multi-institution clinical cooperative trial, 202 eligible patients with a negative SLL were randomly selected to receive either 15 mCi IP 32P (n = 104) or no further therapy (NFT; n = 98). RESULTS: With a median follow-up of 63 months in living patients, 68 patients in the IP 32P group (65%) and 63 patients in the NFT group (64%) have developed tumor recurrence. The relative risk of recurrence is 0.90 (IP 32P to NFT) (90% confidence interval [CI], 0.68 to 1.19). The 5-year RFS rate is 42% and 36% for the IP 32P and NFT groups, respectively; the difference is not statistically significant (log-rank test, P =.27). There was no statistically significant difference in OS (P =.19). The relative risk of death is 0.85 (IP 32P to NFT) (90% CI, 0.62 to 1.16). Sixteen patients (8%) experienced grade 3 or 4 adverse effects, with eight in each respective group. CONCLUSION: Intraperitoneal chromic phosphate did not decrease the risk of relapse or improve survival for patients with stage III epithelial ovarian cancer after a negative SLL. Despite complete pathologic remission at SLL after initial surgery and platinum-based chemotherapy, 61% of stage III ovarian cancer patients had tumor recurrence within 5 years of negative SLL. This indicates a need for more effective initial therapy and further studies of consolidation therapy. PMID- 12885801 TI - Phase II study of oxaliplatin in platinum-resistant and refractory ovarian cancer: a gynecologic group study. AB - PURPOSE: A phase II study was conducted to determine the efficacy of oxaliplatin therapy in patients with platinum-resistant or refractory epithelial ovarian carcinoma. MATERIALS AND METHODS: Eligible patients were to receive oxaliplatin 130 mg/m2 intravenously over 2 hours, every 21 days, until progression of disease or adverse effects prohibited further therapy. RESULTS: Of 25 patients entered onto the study, 23 were eligible and assessable. There were no patients with complete response. One patient (4.3%) achieved a partial response, with a response duration of 6.4 months. Nine patients (39.1%) experienced stable disease, with a median duration of 5.6+ months (range, 1.8 to 13.1 months). The most frequently reported drug-related toxicities were hematologic, gastrointestinal, and neurologic. CONCLUSION: Oxaliplatin as a single agent has minimal activity in patients with platinum-resistant or refractory ovarian cancer at the dosage and schedule tested. However, future studies of oxaliplatin combined with other active agents in women with platinum-naive or platinum sensitive epithelial ovarian carcinoma may be indicated. PMID- 12885802 TI - Prognostic significance of visible lesions on transrectal ultrasound in impalpable prostate cancers: implications for staging. AB - PURPOSE: The current tumor-node metastasis (TNM) staging system classifies impalpable prostate cancers identified by needle biopsy and invisible by imaging as T1c and those visible as T2. Palpable cancers are classified as at least T2. However, most urologists consider impalpable prostate cancers T1c tumors, irrespective of findings on transrectal ultrasound (TRUS). The aim of this article is to provide a differentiated view of the significance of TRUS findings for staging purposes in impalpable prostate cancers. PATIENTS AND METHODS: A consecutive series of 1670 patients with impalpable tumors and palpable T2 cancers after radical prostatectomy were evaluated. Tumor characteristics and 5 year biochemical cure rates of cancers invisible and visible on TRUS were compared, as well as the rates of impalpable but visible and palpable T2 cancers. RESULTS: Impalpable cancers invisible on TRUS presented significantly more favorable pathologic stages and lower cancer volumes than those visible on TRUS (P =.002, P =.010). In the latter, these clinical features were more favorable compared with T2 cancers (P <.001, P <.001). Progression-free probability of impalpable cancers invisible on TRUS was 86.8%; progression-free probability for impalpable cancers visible on TRUS was 85.4% (log-rank test P =.2060). The corresponding rate for T2 tumors was 73.9%, significantly lower when compared to those of visible and impalpable cancers (log-rank test P =.0001). CONCLUSION: Impalpable prostate cancers invisible on TRUS present more favorable cancer features than those that are visible on TRUS. However, these differences are not as pronounced as those between impalpable but visible cancers and palpable T2 tumors. Thus, based on our data, it seems inappropriate to classify impalpable prostate cancers visible on TRUS as T2 cancers. PMID- 12885803 TI - Repeated bone-targeted therapy for hormone-refractory prostate carcinoma: tandomized phase II trial with the new, high-energy radiopharmaceutical rhenium 188 hydroxyethylidenediphosphonate. AB - PURPOSE: We investigated the effect of repeated bone-targeted therapy with rhenium-188 hydroxyethylidenediphosphonate (HEDP) in patients with progressive, hormone-resistant prostate carcinoma and bone pain. The aim of this study was to determine the pain palliation and the antitumor effect of rhenium-188 HEDP treatments. PATIENTS AND METHODS: Sixty-four patients were randomly assigned to one of two groups for radionuclide therapy with rhenium-188 HEDP; patients of group A received a single injection, patients of group B received two injections (interval, 8 weeks). After therapy, patients were followed-up by assessment of pain palliation and clinical outcome until death. RESULTS: In both groups, toxicity was low, with moderate thrombopenia and leukopenia (maximum common toxicity criteria grade of 2). The effectiveness of rhenium-188 HEDP for pain palliation was better in the repeated treatment group (group B), with a response rate and time of response of 92% and 5.66 months, respectively (P =.006 and P =.001). In group B, 11 (39%) of 28 patients had a prostate-specific antigen decrease of more than 50% for at least 8 weeks, compared with two (7%) of 30 patients in the single-injection group (group A). The median times to progression of group A and group B were 2.3 months (range, 0 to 12.2 months) and 7.0 months (range, 0 to 24.1 months), respectively (P =.0013), and the median overall survival times were 7.0 months (range, 1.3 to 36.7 months) and 12.7 months (range, 4.1 to 32.2 months), respectively (P =.043). CONCLUSION: Compared with single-injection therapy, repeated bone-targeted therapy with rhenium-188 HEDP administered to patients with advanced progressive hormone-refractory prostate carcinoma enhanced pain palliation and improved progression-free and overall survival. Larger studies are justified to further evaluate the use of rhenium-188 HEDP. PMID- 12885804 TI - AIDS-related Kaposi's Sarcoma: evaluation of potential new prognostic factors and assessment of the AIDS Clinical Trial Group Staging System in the Haart Era--the Italian Cooperative Group on AIDS and Tumors and the Italian Cohort of Patients Naive From Antiretrovirals. AB - PURPOSE: To assess potential new prognostic factors and to validate the AIDS Clinical Trials Group (ACTG) for AIDS-related Kaposi's sarcoma (AIDS-KS) staging system in the highly active antiretroviral therapy (HAART) era. PATIENTS AND METHODS: We collected epidemiologic, clinical, staging, and survival data from 211 patients with AIDS-KS enrolled in two prospective Italian human immunodeficiency virus (HIV) cohort studies. We included in the analysis all patients with the diagnosis of KS made from January 1996, the time at which HAART became available in Italy. RESULTS: In the univariate analysis, survival was not influenced by sex, age, level of HIV viremia at KS diagnosis, HAART at KS diagnosis (HAART-naive v HAART-experienced), or type of HAART combination. Regarding ACTG classification, the 3-year survival rate was 85% for T0 patients and 69% for T1 patients (P =.007), 83% for S0 patients and 63% for S1 patients (P =.003), and 83% for I0 patients and 71% for I1 patients (P =.06). In the multivariate analysis, only the combination of poor tumor stage (T1) and poor systemic disease (S1) risk identified patients with unfavorable prognosis. The 3 year survival rate of patients with T1S1 was 53%, which was significantly lower compared with the 3-year survival rates of patients with T0S0, T1S0, and T0S1, which were 88%, 80%, and 81%, respectively (P =.0001). CONCLUSION: In the era of HAART, a refinement of the original ACTG staging system is needed. CD4 level does not seem to provide prognostic information. Two different risk categories are identified: a good risk (T0S0, T1S0, T0S1) and a poor risk (T1S1). PMID- 12885805 TI - Prospective randomized trial of interferon alfa-2b and interleukin-2 as adjuvant treatment for resected intermediate- and high-risk primary melanoma without clinically detectable node metastasis. AB - PURPOSE: Low-dose interferon alfa (IFNalpha) has been shown to have limited effects in the adjuvant treatment of patients with intermediate- and high-risk primary melanoma. We hypothesized that a combination regimen with low-dose interleukin-2 (IL-2) may improve survival prospects in these patients. PATIENTS AND METHODS: After wide excision of primary melanoma without clinically detectable lymph node metastasis (pT3 to 4, cN0, M0), 225 patients from 10 participating centers were randomly assigned to receive either subcutaneous low dose IFNalpha2b (3 million international units [MU]/m2/d, days 1 to 7, week 1; three times weekly, weeks 3 to 6, repeated all 6 weeks) plus IL-2 (9 MU/m2/d, days 1 to 4, week 2 of each cycle) for 48 weeks, or observation alone. The primary end point was prolongation of a relapse-free interval. RESULTS: Of the 225 enrolled patients, 223 were found to be eligible. Median follow-up time was 79 months. All evaluated prognostic factors were well balanced between the two arms of the study. Relapses were noticed in 36 of 113 patients treated with IFNalpha2b plus IL-2 and in 34 of 110 patients with observation alone. Five-year disease-free survival of those who had routine surgery supplemented by IFNalpha2b and IL-2 treatment was 70.1% (95% confidence interval [CI], 61.3% to 78.9%), compared with 69.9% in those receiving surgery and observation alone (95% CI, 60.7% to 79.1%) in the intention-to-treat analysis. Evaluation of the overall survival did not show any difference between treated and untreated melanoma patients (P =.93). CONCLUSION: Adjuvant treatment of intermediate- and high-risk melanoma patients with low-dose IFNalpha2b and IL-2 is safe and well tolerated by most patients, but it does not improve disease-free or overall survival. PMID- 12885806 TI - Trastuzumab and vinorelbine as first-line therapy for HER2-overexpressing metastatic breast cancer: multicenter phase II trial with clinical outcomes, analysis of serum tumor markers as predictive factors, and cardiac surveillance algorithm. AB - PURPOSE: Trastuzumab-based therapy improves survival for women with human epidermal growth factor receptor 2 (HER2)-positive advanced breast cancer. We conducted a multicenter phase II study to evaluate the efficacy and safety of trastuzumab combined with vinorelbine, and to assess cardiac surveillance algorithms and tumor markers as prognostic tools. PATIENTS AND METHODS: Patients with HER2-positive (immunohistochemistry [IHC] 3+-positive or fluorescence in situ hybridization [FISH]-positive) metastatic breast cancer received first-line chemotherapy with trastuzumab and vinorelbine to determine response rate. Eligibility criteria were measurable disease and baseline ejection fraction >or= 50%. Serial testing for HER2 extracellular domain (ECD) was performed. RESULTS: Fifty-four women from 17 participating centers were entered onto the study. The overall response rate was 68% (95% confidence interval, 54% to 80%). Response rates were not affected by method of HER2 status determination (FISH v IHC) or by prior adjuvant chemotherapy. Median time to treatment failure was 5.6 months; 38% of patients were progression free after 1 year. Concurrent therapy was quite feasible with maintained dose-intensity. Patients received both chemotherapy and trastuzumab on 90% of scheduled treatment dates. Two patients experienced cardiotoxicity in excess of grade 1; one patient experienced symptomatic heart failure. A surveillance algorithm of screening left ventricular ejection fraction (LVEF) at 16 weeks successfully identified women at risk for experiencing cardiotoxicity. Other acute and chronic side effects were tolerable. Lack of decline in HER2 ECD during cycle 1 predicted tumor progression. CONCLUSION: Trastuzumab and vinorelbine constitute effective and well-tolerated first-line treatment for HER2-positive metastatic breast cancer. Patients with normal LVEF can be observed with surveillance of LVEF at 16 weeks to identify those at risk for cardiotoxicity. PMID- 12885807 TI - Semimonthly versus monthly regimen of fluorouracil and leucovorin administered for 24 or 36 weeks as adjuvant therapy in stage II and III colon cancer: results of a randomized trial. AB - PURPOSE: This randomized, 2 x 2 factorial study compared a semimonthly (LVFU2) with a monthly (FULV) regimen of fluorouracil and leucovorin and 24 versus 36 weeks of each regimen as adjuvant treatment of patients with stage II (Dukes' B2) and III (Dukes' C) colon cancer. PATIENTS AND METHODS: LVFU2 was administered semi-monthly for 2 consecutive days as dl- or l-leucovorin (200 or 100 mg/m2, respectively) as a 2-hour infusion, followed by a 400 mg/m2 FU bolus and 600 mg/m2 of FU as a 22-hour continuous infusion. FULV was administered monthly for 5 consecutive days as a 15-minute infusion of dl- or l-leucovorin, followed by 400 mg/m2 of FU as a 15-minute infusion. RESULTS: A total of 905 patients were randomly assigned. The median follow-up was 41 months. Disease-free survival was similar between the LVFU2 and FULV groups (127 v 124 events; hazard ratio [HR] = 1.04; P =.74) and between 24 and 36 weeks of therapy (128 v 123 events; HR = 0.94; P =.63). Analysis of overall survival showed a slight excess in the number of deaths in LVFU2 compared with FULV (73 v 59), but this difference was not statistically significant (HR = 1.26; 95% confidence interval, 0.90 to 1.78; P =.18). The most commonly observed grade 3 to 4 toxicities were neutropenia, diarrhea, and mucositis. Toxicities were significantly lower in the LVFU2 group (all toxicities, P <.001). CONCLUSION: Our data confirm that LVFU2 is less toxic than FULV. At a median follow-up of 41 months, no statistically significant difference could be detected in disease-free or overall survival between the treatment groups or treatment durations. PMID- 12885808 TI - Safety and toxicity analysis of oxaliplatin combined with fluorouracil or as a single agent in patients with previously treated advanced colorectal cancer. AB - PURPOSE: Two consecutive compassionate use studies of oxaliplatin were conducted in the United States and Canada in more than 5000 patients with locally advanced or metastatic colorectal carcinoma who had experienced treatment failure after at least one prior chemotherapy regimen. PATIENTS AND METHODS: The main focus was safety. Patients were assigned to treatment with either single-agent oxaliplatin or oxaliplatin in combination with fluorouracil (FU) and with or without leucovorin (LV) in various regimens. Response data collection was not a trial objective, but time to treatment failure (TTF) was recorded in the first cohort (1370 patients). RESULTS: All treatment regimens were well tolerated, with an overall incidence of grade 3 or 4 hematologic toxicity of 23.2%, grade 3 or 4 treatment-related gastrointestinal toxicity of 26.4% (including diarrhea, vomiting, and mucositis), and grade 3 neurosensory toxicity 3.9%. Similar results were reported in the second cohort (3806 patients), in which the eligibility criteria were much less restrictive. In the first cohort (in which 83% received prior irinotecan), median TTF was 14 weeks, and was similar for the five regimens combining oxaliplatin and FU with or without LV, but significantly shorter for the single-agent oxaliplatin arm. The overall dose-intensity of oxaliplatin was maintained at 85.5% (range, 80.6% to 94.3%) of that prescribed by protocol (average 36.7 mg/m2/wk). CONCLUSION: These data in a heavily pretreated patient population confirm that oxaliplatin is safe when used as a single agent or with a variety of FU-based regimens as salvage therapy in patients with advanced colorectal cancer. PMID- 12885809 TI - Colon cancer survival is associated with increasing number of lymph nodes analyzed: a secondary survey of intergroup trial INT-0089. AB - PURPOSE: To determine the relationship, in patients with adenocarcinoma of the colon, between survival and the number of lymph nodes analyzed from surgical specimens. PATIENTS AND METHODS: Intergroup Trial INT-0089 is a mature trial of adjuvant chemotherapy for high-risk patients with stage II and stage III colon cancer. We performed a secondary analysis of this group with overall survival (OS) as the main end point. Cause-specific survival (CSS) and disease-free survival were secondary end points. Rates for these outcome measures were estimated using Kaplan-Meier methodology. Log-rank test was used to compare overall curves, and Cox proportional hazards regression was used to multivariately assess predictors of outcome. RESULTS: The median number of lymph nodes removed at colectomy was 11 (range, one to 87). Of the 3411 assessable patients, 648 had no evidence of lymph node metastasis. Multivariate analyses were performed on the node-positive and node-negative groups separately to ascertain the effect of lymph node removal. Survival decreased with increasing number of lymph node involvement (P =.0001 for all three survival end points). After controlling for the number of nodes involved, survival increased as more nodes were analyzed (P =.0001 for all three end points). Even when no nodes were involved, OS and CSS improved as more lymph nodes were analyzed (P =.0005 and P =.007, respectively). CONCLUSION: The number of lymph nodes analyzed for staging colon cancers is, itself, a prognostic variable on outcome. The impact of this variable is such that it may be an important variable to include in evaluating future trials. PMID- 12885810 TI - Phase II study of gemcitabine, cisplatin, and infusional fluorouracil in advanced pancreatic cancer. AB - PURPOSE: This phase II study was undertaken to determine the efficacy of adding infusional fluorouracil (FU) to the chemotherapy doublet of gemcitabine and cisplatin in patients with advanced pancreatic cancer. PATIENTS AND METHODS: The eligibility criteria included histologically or cytologically confirmed adenocarcinoma of the pancreas that was either unresectable or metastatic. No prior gemcitabine therapy was allowed. Patients received a combination of gemcitabine 1000 mg/m2 intravenously (IV) on days 1, 8, and 15; cisplatin 50 mg/m2 IV on days 1 and 15; and FU 175 mg/m2/d from days 1 to 15 by continuous IV infusion. Cycles were repeated every 28 days. Objective tumor response and toxicity were evaluated according to the World Health Organization criteria. RESULTS: A total of 47 patients (median age, 57 years; males, 59%) were enrolled. Sixteen patients had locally advanced (LA) disease, and 31 patients had metastatic disease. A total of 183 cycles of chemotherapy were administered. In patients with metastatic disease (n = 31), the probability of survival at 6 and 12 months was 66% and 34%, respectively. Objective partial response or stable disease was observed in 26% (90% confidence interval [CI], 0.14 to 0.41) and 61% (90% CI, 0.45 to 0.74) of patients, respectively. In patients with LA disease (n = 16), there were three partial responses (19%; 90 CI, 0.07 to 0.39). One patient in this group was successfully resected after FU-based radiotherapy. The most common grade 3 to 4 toxicities were neutropenia (60%), thrombocytopenia (42%), and anemia (26%). Thirteen patients were hospitalized for treatment-related complications. CONCLUSION: The combination of gemcitabine, cisplatin, and infusional FU has significant activity in patients with advanced pancreatic cancer. PMID- 12885811 TI - Phase I trial of escalating-dose irinotecan given weekly with cisplatin and concurrent radiotherapy in locally advanced esophageal cancer. AB - PURPOSE: To identify the maximum-tolerated dose and dose-limiting toxicity (DLT) of weekly irinotecan combined with cisplatin and radiation in esophageal cancer. PATIENTS AND METHODS: Nineteen patients with clinical stage II to III esophageal squamous cell or adenocarcinoma were treated on this phase I trial. Induction chemotherapy with weekly cisplatin 30 mg/m2 and irinotecan 65 mg/m2 was administered for four treatments during weeks 1 to 5. Radiotherapy was delivered weeks 8 to 13 in 1.8-Gy daily fractions to a dose of 50.4 Gy. Cisplatin 30 mg/m2 and escalating-dose irinotecan (40, 50, 65, and 80 mg/m2) were administered on days 1, 8, 22, and 29 of radiotherapy. DLT was defined as a 2-week delay in radiotherapy for grade 3 to 4 toxicity. RESULTS: Minimal toxicity was observed during chemoradiotherapy, with no grade 3 or 4 esophagitis, diarrhea, or stomatitis. DLT caused by myelosuppression was seen in two of six patients treated at the 80-mg/m2 dose level, thus irinotecan 65 mg/m2 was defined as the recommended phase II dose. Dysphagia improved or resolved after induction chemotherapy in 13 (81%) of 16 patients who reported dysphagia before therapy. Only one patient (5%) required a feeding tube. Six complete responses (32%) were observed, including four pathologic complete responses in 15 patients selected to undergo surgery (27%). CONCLUSION: Cisplatin, irinotecan, and concurrent radiotherapy can be administered on a convenient schedule with relatively minimal toxicity and an acceptable rate of complete response in esophageal cancer. Further phase II evaluation of this regimen is ongoing. A phase III comparison to fluorouracil or taxane-containing chemoradiotherapy should be considered. PMID- 12885812 TI - Multicenter, randomized trial for stage IIIB or IV non-small-cell lung cancer using weekly paclitaxel and carboplatin followed by maintenance weekly paclitaxel or observation. AB - PURPOSE: To explore the efficacy and safety of three regimens of weekly paclitaxel plus carboplatin as initial therapy and the feasibility of subsequent maintenance therapy versus observation in patients with advanced non-small-cell lung cancer (NSCLC). PATIENTS AND METHODS: Four hundred one patients were randomly assigned to one of the following arms: arm 1, paclitaxel 100 mg/m2 weekly for 3 of 4 weeks with carboplatin (area under the curve [AUC] = 6) on day 1; arm 2, paclitaxel 100 mg/m2 and carboplatin (AUC = 2) weekly for 3 of 4 weeks; or arm 3, paclitaxel 150 mg/m2 cycle 1 and 100 mg/m2 cycle 2 and carboplatin (AUC = 2) weekly for 6 of 8 weeks. Patients who responded (n = 130) at week 16 were randomly assigned to either weekly paclitaxel therapy (70 mg/m2, 3 of 4 weeks; n = 65) or observation (n = 65). RESULTS: For the 390 assessable patients, the objective response rates observed with initial therapy were 32% for arm 1, 24% for arm 2, and 18% for arm 3. The median time to progression and median survival times were 30 and 49 weeks for arm 1, 21 and 31 weeks for arm 2, and 27 and 40 weeks for arm 3, respectively. The 1-year survival rates were 47% for arm 1, 31% for arm 2, and 41% for arm 3. CONCLUSION: Arm 1, paclitaxel 100 mg/m2 weekly for 3 of 4 weeks with carboplatin (AUC = 6) administered on day 1, demonstrates the most favorable therapeutic index in patients with advanced NSCLC. PMID- 12885813 TI - Mitoxantrone and cytarabine induction, high-dose cytarabine, and etoposide intensification for pediatric patients with relapsed or refractory acute myeloid leukemia: Children's Cancer Group Study 2951. AB - PURPOSE: To evaluate the response rate, survival, and toxicity of mitoxantrone and cytarabine induction, high-dose cytarabine and etoposide intensification, and further consolidation/maintenance therapies, including bone marrow transplantation, in children with relapsed, refractory, or secondary acute myeloid leukemia (AML). To evaluate response to 2-chlorodeoxyadenosine (2-CDA) and etoposide (VP-16) in patients who did not respond to mitoxantrone and cytarabine. PATIENTS AND METHODS: Patients with relapsed/refractory AML (n = 101) and secondary AML (n = 13) were entered. RESULTS: Mitoxantrone and cytarabine induction achieved a remission rate of 76% for relapsed/refractory patients and 77% for patients with secondary AML, with a 3% induction mortality rate. Cytarabine and etoposide intensification exceeded the acceptable toxic death rate of 10%. The response rate of 2-CDA/VP-16 was 8%. Two-year overall survival was estimated at 24% and was better than historical control data. Patients with secondary AML had similar outcomes to relapsed or refractory patients. Initial remission longer than 1 year was the most important prognostic factor for patients with primary AML (2-year survival rate, 75%), whereas for patients with primary AML, with less than 12 months of initial remission, survival was 13% and was similar to that of refractory patients (6%). CONCLUSION: Mitoxantrone and cytarabine induction is effective with reasonable toxicity in patients with relapsed/refractory or secondary AML. The cytarabine and etoposide intensification regimen should be abandoned because of toxicity. Patients with relapsed AML with initial remissions longer than 1 year have a relatively good prognosis. PMID- 12885814 TI - Lymphocyte-predominant Hodgkin's lymphoma in children: therapeutic abstention after initial lymph node resection--a Study of the French Society of Pediatric Oncology. AB - PURPOSE: To clarify treatment strategy for lymphocyte-predominant Hodgkin's lymphoma (LPHL), the French Society of Pediatric Oncology initiated a prospective, nonrandomized study in 1988. Patients received either standard treatment for Hodgkin's lymphoma or were not treated beyond initial adenectomy. PATIENTS AND METHODS: From 1988 to 1998, 27 patients were available for study. Twenty-four patients were male, and median age was 10 years (range, 4 to 16 years). Twenty-two, two, and three patients had stage I, II, and III disease, respectively. Thirteen patients (stage I, n = 11; stage III, n = 2) received no further treatment after initial surgical adenectomy (SA). Fourteen patients received combined treatment (CT; n = 10), involved-field radiotherapy alone (n = 1), or chemotherapy alone (n = 3). The two groups were comparable for clinical status, treatment, and follow-up. RESULTS: Twenty-three of 27 patients achieved complete remission (CR). With a median follow-up time of 70 months (range, 32 to 214 months), overall survival to date is 100%, and overall event-free survival (EFS) is 69% +/- 10% (SA, 42% +/- 16%; CT, 90% +/- 8.6%; P <.04). If we considered only the patients in CR after initial surgery (n = 12), EFS was no longer significantly different between the two groups. Patients with residual mass after initial surgery (n = 15) had worse EFS if they did not receive complementary treatment (P <.05). CONCLUSION: Although based on a small number of patients, our study showed that (1). no further therapy is a valid therapeutic approach in LPHL patient in CR after initial lymph node resection, and (2). complementary treatment diminishes relapse frequency but has no impact on survival. PMID- 12885815 TI - Height and weight in children treated for acute lymphoblastic leukemia: relationship to CNS treatment. AB - PURPOSE: We evaluated the long-term effects of treatment on height and weight in children with acute lymphoblastic leukemia (ALL) treated with one of the following three different CNS therapies: intrathecal therapy alone, intrathecal therapy with conventional cranial radiation, or intrathecal therapy with twice daily radiation. PATIENTS AND METHODS: Between 1987 and 1995, 618 children treated on two consecutive Dana-Farber Cancer Institute Consortium protocols for ALL were measured for height and weight at diagnosis, and approximately every 6 months thereafter. Patient height, weight, and body mass index (BMI) were converted to z scores for age and sex using the 2000 Centers for Disease Control and Prevention growth charts for the United States. RESULTS: Children younger than 13 years at diagnosis had a statistically significant decrease in their height z scores and an increase in their BMI z scores, regardless of whether they had received cranial radiation. Young age at diagnosis and increased chemotherapy intensity were major risk factors. Unexpectedly, there was no significant difference in long-term height between children who received radiation and those who did not. CONCLUSION: Final height is compromised in survivors of ALL. The detrimental effects on height occur during therapy without the ability for long term catch-up growth. Although patients became overweight for height, this seemed to be a result of relative height loss with normal weight gain rather than accelerated weight gain. The type of CNS treatment received did not affect changes in height, weight, or BMI. PMID- 12885816 TI - Cognitive outcome of long-term survivors of multisystem langerhans cell histiocytosis: a single-institution, cross-sectional study. AB - PURPOSE: Damage to the CNS, including the cerebellum, and to the hypothalamopituitary axis, is documented in Langerhans cell histiocytosis (LCH). Neuropsychologic deficits have been recognized, but this is the first study in which cognitive function has been systematically assessed in a cohort of patients. PATIENTS AND METHODS: Twenty-eight long-term survivors of multisystem LCH (mean age, 15.1 years) were investigated for intelligence, memory and learning, language, and academic attainments. RESULTS: The mean intelligence quotient (IQ) of the entire group was not significantly different from the mean of the population (ie, mean +/- SD, 100 +/- 1), but there were wide ranges (Full Scale IQ [FSIQ]: mean, 93.6; range, 61.7 to 134; Performance IQ [PIQ]: mean, 92.2; range, 46 to 136; and Verbal IQ [VIQ]: mean, 93.7; range, 64.2 to 126). CNS involvement was a significant risk factor for lower scores, but sex, diabetes insipidus, and cranial radiotherapy were not. The CNS group had lower VIQ, PIQ, and FSIQ than patients with no CNS involvement (no CNS group: mean +/- SD FSIQ, 102.3 +/- 15.6; CNS group: mean +/- SD FSIQ, 73.6 +/- 7.7; P <.001). A similar pattern of results was obtained for all other cognitive measures. Even when effects of reduction in FSIQ were taken into account, specific deficits were found in patients in the CNS group. CONCLUSION: Long-term survivors of multisystem LCH, particularly patients with CNS involvement, may develop significant cognitive deficits. All patients should have formal, repeated neuropsychologic assessment as part of long-term follow-up, which will enable abnormalities to be detected early so that appropriate supportive measures can be offered. PMID- 12885817 TI - Study of the MIB-1 labeling index as a predictor of tumor progression in pilocytic astrocytomas in children and adolescents. AB - PURPOSE: The pilocytic astrocytoma (PA) is the most common childhood brain tumor. This report examines the MIB-1 labeling index (LI) as a predictor of progression free survival (PFS) among childhood PAs. PATIENTS AND METHODS: Consecutive PAs were examined to determine whether the MIB-1 LI was associated with tumor progression. Other variables evaluated included tumor location, use of adjuvant therapy, extent of resection, and age at diagnosis. RESULTS: One hundred forty one children were identified (mean +/- SD age, 7.6 +/- 4.7 years; range, 0.43 to 18.56 years); 118 children had adequate tissue for MIB-1 immunohistochemistry. The 5-year PFS was 61.25%. By log-rank analysis, an MIB-1 LI of more than 2.0 was associated with shortened PFS (P =.035). Patients with PAs who underwent complete surgical resection, had tumors located in the cerebellum, and were treated with surgery only also had more prolonged PFS (P =.001 for all). Tumors in the optic pathways were associated with a shorter PFS (P =.001). Restricting the evaluation of MIB-1 LI to only incompletely resected tumors revealed an insignificant trend of MIB-1 LI of more than 2.0 having a shortened PFS. Multivariate analysis demonstrated completely resected tumors and tumors located in the cerebellum as less likely to progress (P =.001 and.019, respectively). CONCLUSION: Children with PAs with an MIB-1 LI of more than 2.0 have a shortened PFS. PAs that are completely resected and are located in the cerebellum have a prolonged PFS. This initial study suggests that the MIB-1 LI identifies a more aggressive subset of PAs. Further work should focus on elucidating features of pilocytic astocytomas that will identify prospectively children at risk for progression. PMID- 12885818 TI - Single center experience of a new intensive induction therapy for ewing's family of tumors: feasibility, toxicity, and stem cell mobilization properties. AB - PURPOSE: To examine the feasibility, tolerability, and toxicity of an intensified induction regimen (vincristine, ifosfamide, doxorubicin, and etoposide [VIDE]) in patients with newly diagnosed Ewing's family of tumors (EFT); to assess ability to maintain dose-intensity, and predictability of peripheral-blood stem cell mobilization. PATIENTS AND METHODS: Thirty patients were treated with vincristine 1.4 mg/m2 (maximum 2 mg) on day 1, doxorubicin 20 mg/m2, ifosfamide 3 g/m2 plus mesna and etoposide 150 mg/m2 on days 1 to 3. Cycles were given every 21 days for up to six cycles. RESULTS: One-hundred and seventy cycles of VIDE were given. The median treatment interval was 21 days (21 to 42) and nadir count: hemoglobin 8.3 (6.3 to 11.9), neutrophils 0.045 (0.0 to 2.1), and platelets 45 (3 to 343). There were 96 episodes of infection requiring hospitalization (56%). Growth factor support reduced infectious complications by 34%. Etoposide dose was reduced, or omitted, in 24% of cycles. Four patients did not complete six cycles due to unacceptable toxicity and one patient progressed on treatment. Twenty patients underwent peripheral-blood stem cell harvesting, 15 after cycle 3, and five after cycle 4. Median CD34+ yield was 4.6 x 106/kg per patient (1.8 to 14.5). Overall response to treatment, measured in 24 patients, was 88%. Seven of 11 patients undergoing surgery achieved greater than 90% necrosis of tumor (64%). CONCLUSION: VIDE is an effective induction regimen with substantial but acceptable toxicity that allows predictable mobilization of stem cells. Maintenance of dose-intensity is feasible in the majority of patients. Growth factors play a role in maintaining dose-intensity and reduce infectious complications. PMID- 12885819 TI - Health-related quality of life in non-small-cell lung cancer: methodologic issues in randomized controlled trials. AB - PURPOSE: Non-small-cell lung cancer (NSCLC), a leading cause of cancer-related deaths in both developed and developing countries, heavily impacts patient health related quality of life (HRQOL). Although recent research has found many positive and significant steps in randomized controlled trials (RCTs) in which HRQOL has been used as an outcome, questions remain regarding methodologic quality and impact value of HRQOL outcome assessments in some RCTs. To date, no detailed systematic review exists of HRQOL methodology in NSCLC RCTs. METHODS: A systematic review using Cochrane methodology evaluated HRQOL components in RCTs. Identified studies were evaluated independently by three reviewers on a broad set of predetermined criteria. RESULTS: Twenty-nine published RCTs (NSCLC patient samples total, N = 8445) with an HRQOL component were identified. Although most trials exhibited good-quality research and useful HRQOL and clinical data, the weaknesses identified in some trials include such common limitations as no clear hypothesis, lack of a clear approach to missing data and data analysis, and limited presentation of results. CONCLUSION: Although it is clear that HRQOL is an important end point in NSCLC RCTs because the information helps to influence treatment recommendations, the identified weaknesses in conducting HRQOL measurement in NSCLC RCTs and the reporting of results need to be addressed. PMID- 12885820 TI - Extraoular muscle palsies in subcutaneous panniculitis-like T-cell lymphoma. PMID- 12885821 TI - Carcinoid tumor presenting with bone marrow metastases. PMID- 12885822 TI - Brain metastasis from ovarian cancer. PMID- 12885823 TI - Tumor implantation after pneumonectomy for lung cancer. PMID- 12885824 TI - Allowing patients to die: practical, ethical, and religious concerns. PMID- 12885826 TI - Alternatives to cystectomy in muscle-invasive bladder cancer. PMID- 12885828 TI - Morphologic dysplasia in acute myeloid leukemia: importance of granulocytic dysplasia. PMID- 12885830 TI - Opioid switching. PMID- 12885831 TI - Will identifying or targeting altered marker expression in response to cytotoxic therapy be of prognostic or therapeutic value? PMID- 12885832 TI - It's what's inside that counts. PMID- 12885833 TI - Down syndrome and acute myeloid leukemia: the paradox of increased risk for leukemia and heightened sensitivity to chemotherapy. PMID- 12885834 TI - Dynamic expression profile of p21WAF1/CIP1 and Ki-67 predicts survival in rectal carcinoma treated with preoperative radiochemotherapy. AB - PURPOSE: We investigated p53 and its downstream effectors p21WAF1/CIP1, BAX, and hMSH2 as well as the proliferation marker Ki-67 (mki-67/MIB-1) in patients undergoing preoperative radiochemotherapy for rectal carcinoma to identify prognostic and predictive factors. The focus of this study was on the dynamics of these genetic markers in a longitudinal study-that is, before and after radiochemotherapy. PATIENTS AND METHODS: Expression of p53, BAX, p21WAF1/CIP1, Ki 67, and hMSH2 was investigated by immunohistochemistry in pre- and posttherapeutic tumor samples in 66 patients. Tumor DNA was screened for p53 mutations by single-strand conformation polymorphism-polymerase chain reaction (SSCP-PCR). Paired tumor samples (pretherapy and posttherapy) were collected prospectively. RESULTS: Patients with a decrease in p21 expression following radiochemotherapy had better disease-free survival (P =.03). Similarly, patients with an increase in proliferative activity as measured by increased Ki-67 expression posttherapy had better disease-free survival (P <.005). In addition, we observed a significantly better prognosis for patients with high hMSH2 expression. In contrast, pretherapeutic levels of p53, BAX, or p21 expression and p53 mutation had no prognostic value, indicating that the combination of radiotherapy and chemotherapy might override defects in these genes. CONCLUSION: These findings are novel and support the clinical relevance of p21 in the suppression of both proliferation and apoptosis. Thus, the dynamic induction of p21WAF1/CIP1 was associated with a lower proliferative activity but an ultimately worse treatment outcome following neoadjuvant radiochemotherapy and tumor resection. Induction of p21, therefore, represents a novel resistance mechanism in rectal cancer undergoing preoperative radiochemotherapy. PMID- 12885835 TI - Long-term cause-specific mortality of patients treated for Hodgkin's disease. AB - PURPOSE: To assess long-term cause-specific mortality of young Hodgkin's disease (HD) patients. PATIENTS AND METHODS: The study population consisted of 1,261 patients treated for HD before age 41 between 1965 and 1987. Follow-up was complete until October 2000. For 95% of deaths, the cause was known. Long-term cause-specific mortality was compared with general population rates to assess relative risk (RR) and absolute excess risk (AER) of death. RESULTS: After a median follow-up of 17.8 years, 534 patients had died (55% of HD). The RR of death from all causes other than HD was 6.8 times that of the general population, and still amounted to 5.1 after more than 30 years. RRs of death resulting from solid tumors (STs) and cardiovascular disease (CVD) were increased overall (RR = 6.6 and 6.3, respectively), but especially in patients treated before age 21 (RR = 14.8 and 13.6, respectively). When these patients grew older, this elevated mortality decreased. The overall AER of death from causes other than HD increased throughout follow-up. Patients receiving salvage chemotherapy had a significantly increased RR of death from STs, compared to patients receiving initial therapy only. CONCLUSION: The main cause of death among HD patients was lymphoma, but after 20 years, HD mortality was negligible. The RRs and AERs of death from second primary cancers (SCs) and CVDs continued to increase after 10 years. Even more than 30 years after diagnosis, HD patients experienced elevated risk of death from all causes other than HD. Increased risk of death from SCs and CVDs was found especially in patients treated before age 21, but these risks seemed to abate with age. PMID- 12885836 TI - Increased age at diagnosis has a significantly negative effect on outcome in children with Down syndrome and acute myeloid leukemia: a report from the Children's Cancer Group Study 2891. AB - PURPOSE: To determine the outcome of children with Down syndrome (DS) and acute myeloid leukemia (AML) receiving standard timing chemotherapy without bone marrow transplantation (BMT), with determination of prognostic factors. PATIENTS AND METHODS: Children with DS and newly diagnosed AML or myelodysplasia were prospectively enrolled on Children's Cancer Group study 2891 (N = 161) and treated uniformly with four standard timing induction courses of dexamethasone, cytarabine arabinoside, 6-thioguanine, etoposide, daunorubicin (DCTER) followed by intensively timed high-dose cytarabine. RESULTS: Children with DS were significantly younger at diagnosis than those without (median age, 1.8 v 7.5 years, respectively; P <.001), with more megakaryocytic leukemia (70% v 6%; P <.001). Higher complete remission rates (91%) were achieved in children with DS than among those without DS (75%; P <.001). Equivalent grade 3 to 4 toxicity (29% v 30%; P =.84) was seen, though children with DS had greater pulmonary toxicity (P <.01) during induction and mucositis during intensification (P =.12). Children with DS had significantly better 8-year event-free survival (EFS; 77% v 21% standard and 40% intensive induction; P <.0001). Multivariate analysis in children with DS revealed that only age at diagnosis of 2 years or older was a risk factor for greater relapse risk (odds ratio, 4.9; P =.006) and worse survival. Children between ages 0 to 2 years (n = 94) had a 6-year EFS of 86%; those from 2 to 4 years (n = 58), 70%; and those older than 4 years (n = 9), 28%. Remission failures were the primary reason for worse 6-year EFSs (1% in those 0 to 2 years v 14% if >2 years; P =.002). CONCLUSION: Outcome for children with DS and AML is excellent with standard induction therapy, but declines with increasing age. PMID- 12885838 TI - It's never all black and white. PMID- 12885837 TI - Randomized phase II comparison of dose-intense gemcitabine: thirty-minute infusion and fixed dose rate infusion in patients with pancreatic adenocarcinoma. AB - PURPOSE: To conduct a randomized phase II trial of dose-intense gemcitabine using a standard 30-minute infusion or the fixed dose rate (FDR) infusion (10 mg/m2/min) in patients with pancreatic adenocarcinoma. PATIENTS AND METHODS: In this prospective trial, patients with locally advanced and metastatic pancreatic adenocarcinoma were treated with 2,200 mg/m2 gemcitabine over 30 minutes (standard arm) or 1,500 mg/m2 gemcitabine over 150 minutes (FDR arm) on days 1, 8, and 15 of every 4-week cycle. The primary end point of this trial was time to treatment failure. Secondary end points included time to progression, median survival, safety, and pharmacokinetic studies of gemcitabine. RESULTS: Ninety-two patients were enrolled onto this study; 91% of the patients had metastatic disease. Time to treatment failure was comparable in both treatment groups; however, the median survival for all patients was 5.0 months in the standard arm and 8.0 months in the FDR arm (P =.013). For patients with metastases, the median survival was 4.9 months in the standard arm and 7.3 months in FDR arm (P =.094). The 1- and 2-year survival rates for all patients were 9% (standard arm) versus 28.8% (FDR; P =.014) and 2.2% (standard arm) versus 18.3% (FDR; P =.007), respectively. Patients in the FDR infusion arm experienced consistently more hematologic toxicity. Pharmacokinetic analyses demonstrated a two-fold increase in intracellular gemcitabine triphosphate concentration in the FDR arm (P =.046). CONCLUSION: Pharmacokinetic and clinical data in this trial supports the continued evaluation of the FDR infusion strategy with gemcitabine. PMID- 12885839 TI - Bone with a vascular flap induced from fat tissue with the use of rhBMP-2 in rats. AB - Here we report that successful bone formation with a vascular flap inside a cylindrical mold was induced from fat tissue with the use of recombinant human bone morphogenetic protein-2 in rats. Fat tissue connected to blood vessels was prepared to fit into the mold and implanted intramuscularly into the hind leg in Wistar rats. RhBMP-2 (20 micro g) was applied in a collagen sheet previously placed on the inside surface of the mold. Bone formation was confirmed radiologically and morphologically at 2, 4, and 8 weeks after the surgery. In the control group without rhBMP-2 or the group with ligation of the blood vessels before the implantation, bone formation was not observed. Our success in bone formation having a definite size, shape, and blood supply may lead to a therapeutic approach to effective bone reconstitution. The present study is the first report on bone induction from fat tissue by rhBMP-2 in vivo. PMID- 12885840 TI - Effects of implant healing time on crestal bone loss of a controlled-load dental implant. AB - The universally accepted concept of delay-loaded dental implants has recently been challenged. This study hypothesizes that early loading (decreased implant healing time) leads to increased bone formation and decreased crestal bone loss. We used 17 minipigs to study implants under a controlled load, with non-loaded implants for comparison. Radiographic and histological assessments were made of the osseointegrated bone changes for 3 healing times (between implant insertion and loading), following 5 months of loading. The effect of loading on crestal bone loss depended on the healing time. Early loading preserved the most crestal bone. Delayed loading had significantly more crestal bone loss compared with the non-loaded controls (2.4 mm vs. 0.64 mm; P < 0.05). The histological assessment and biomechanical analyses of the healing bone suggested that loading and bioactivities of osteoblasts exert a synergistic effect on osseointegration that is likely to support the hypothesis that early loading produces more favorable osseointegration. PMID- 12885841 TI - Adhesive resin induces apoptosis and cell-cycle arrest of pulp cells. AB - The application of an adhesive resin near or directly over the pulp was shown to induce pulp inflammation and lack of dentin regeneration. We hypothesize that the absence of dentin bridging is due to adhesive-resin-induced apoptosis of cells responsible for pulp healing and dentin regeneration. Mouse odontoblast-like cells (MDPC-23), undifferentiated pulp cells (OD-21), or macrophages (RAW 264.7) were exposed to SingleBond polymerized for 0-40 seconds. Annexin V and propidium iodide assays demonstrated that SingleBond induced apoptosis of MDPC-23, OD-21, and macrophages. The proportion of apoptotic cells was dependent on the degree of adhesive resin polymerization. Adhesive-resin-induced death of pulp cells was associated with activation of the pro-apoptotic cysteine protease Caspase-3. Interestingly, most cells exposed to adhesive resin that did not undergo apoptosis showed cell-cycle arrest. We conclude that an adhesive resin induces apoptosis and cell-cycle arrest of cells involved in the regeneration of the dentin-pulp complex in vitro. PMID- 12885842 TI - Effects of HEMA/solvent combinations on bond strength to dentin. AB - Re-expansion of dried demineralized dentin is required to optimize resin adhesion. This study tested the hypothesis that bond strengths to dentin depend upon the ability of experimental HEMA(2-hydroxy-ethyl-methacrylate)/solvent primers to re-expand the matrix. Dentin surfaces were acid-etched with 37% phosphoric acid for 20 sec, air-dried for 30 sec, primed with either 35/65% (v/v) HEMA/water, HEMA/methanol, HEMA/ethanol, or HEMA/propanol for 60 sec, and bonded with 4-META-TBBO(4-methacryloyloxyethyl trimellitate anhydride-tri-n-butyl borane) adhesive. After storage in water for 1 day at 37 degrees C, the samples were prepared for microtensile bond strength testing. We used transmission electron microscopy to measure the width of interfibrillar spaces in the hybrid layers. The HEMA/ethanol primer and the HEMA/propanol primer produced the highest and the lowest bond strengths, respectively (p < 0.05). Bond strengths were directly correlated with the width of the interfibrillar spaces (p < 0.05). Bond strengths are related to the ability of the primer to maintain the re-expansion of collapsed demineralized dentin matrix. PMID- 12885843 TI - Artifacts from dental casting alloys in magnetic resonance imaging. AB - The potential advantage of magnetic resonance imaging (MRI) has been limited by artifacts due to the presence of metallic materials. For quantitative evaluation of the magnitude of artifacts from dental casting alloys and implant materials in MR imaging, 11 dental casting or implant materials were imaged by means of 1.5 T MRI apparatus with three different sequences. Mean and standard deviation of water signal intensity (SI) around the sample in the region of interest (1200 mm(2)) were determined, and the coefficient of variation was compared for evaluation of the homogeneity of the SI. A variety of artifacts with different magnitudes was observed. Only one of the samples, composed mainly of Pd, In, and Sb, showed no artifacts in all imaging sequences. We concluded that selection of specific dental casting alloys according to their elemental compositions could minimize the metal artifacts in MRI; however, titanium alloys currently pose a problem with respect to causing MRI artifacts. PMID- 12885844 TI - Expression of an inhibitor of apoptosis, survivin, in oral carcinogenesis. AB - A novel inhibitor of apoptosis, survivin, plays a role in oncogenesis. To determine the potential involvement of survivin in oral carcinogenesis, we investigated the distribution of survivin protein expression in oral squamous cell carcinomas (OSCCs) and oral pre-malignant lesions. The mRNA expression level and methylation status of the gene also were evaluated in OSCCs and OSCC-derived cell lines. In immunohistochemistry, 58% of tumors and 37% of pre-malignant lesions examined were positive for survivin, while no immunoreaction was observed in corresponding normal tissues. The reverse-transcription/polymerase chain reaction revealed similar changes in survivin gene expression levels. Furthermore, of the 9 normal oral tissues with no survivin gene expression, 4 showed methylation of the gene, while no methylation was detected in the corresponding tumorous tissues. The results suggest that survivin plays an important role during oral carcinogenesis, and that the gene expression may be regulated by an epigenetic mechanism. PMID- 12885845 TI - Single-nucleotide polymorphism in the CD14 promoter and periodontal disease expression in a Japanese population. AB - It has been reported that there is a relationship between a single-nucleotide polymorphism (SNP) in the promoter region of the CD14 gene at position -159 (C- >T) and infectious diseases. The aim of the present study was to test the hypthesis that expression of this SNP correlates with periodontal disease in a Japanese population. The CD14 genotype was determined in 163 subjects with periodontitis and in 104 age- and gender-matched control subjects without periodontitis. The genotype distribution and allele frequency within the periodontitis patients were not significantly different from those of control subjects. There was, however, a significant difference in the genotype distribution between young patients (< 35 yrs) and older patients (>/==" BORDER="0"> 35 yrs). These findings suggest that CD14 -159C/T polymorphism is not related to the development of periodontitis in a Japanese population, but that, within the periodontitis subjects, expression of the SNP may be related to early disease activity. PMID- 12885846 TI - Task-dependence of jaw elevator and depressor co-activation. AB - Elevator muscle activity per unit bite-force has been shown to be higher in chewing than in isometric biting. We tested the hypothesis that surplus elevator activity is evoked in response to a possible co-activation of jaw-opener muscles during the masticatory power stroke. In 32 subjects, digastric and bilateral masseter and temporalis activities were recorded during unilateral chewing of test foods, isometric biting on a force transducer, and during balancing of the jaw against maximum effort of depressor muscles. During elevator peak effort in chewing, the digastric activity was 113% higher than during peak effort in isometric biting. Comparison of balancing and chewing trials revealed that a 6% increase of elevator activity would suffice to compensate for this increased depressor action. Elevator activity in chewing, however, was up to 130% higher than in clenching. We conclude that depressor counteraction could have only a minor influence on the generation of surplus muscle activity in chewing. PMID- 12885848 TI - PTHrP regulates chondrocyte maturation in condylar cartilage. AB - PTHrP is a key factor regulating the pace of endochondral ossification during skeletal development. Mandibular advancement solicits a cascade of molecular responses in condylar cartilage. However, the pace of cellular maturation and its effects on condylar growth are still unknown. The purpose of this study was to evaluate the pattern of expression of PTHrP and correlate it to cellular dynamics of chondrocytes in condylar cartilage during natural growth and mandibular advancement. We fitted 35-day-old Sprague-Dawley rats with functional appliances. Experimental animals with matched controls were labeled with bromodeoxyuridine 3 days before their death, so that mesenchymal cell differentiation could be traced. Mandibular advancement increased the number of differentiated chondroblasts and subsequently increased the cartilage volume. Higher levels of PTHrP expression in experimental animals coincided with the slowing of chondrocyte hypertrophy. Thus, mandibular advancement promoted mesenchymal cell differentiation and triggered PTHrP expression, which retarded their further maturation to allow for more growth. PMID- 12885847 TI - Differential injury responses in oral mucosal and cutaneous wounds. AB - Oral mucosa heals faster than does skin, yet few studies have compared the repair at oral mucosal and cutaneous sites. To determine whether the privileged healing of oral injuries involves a differential inflammatory phase, we compared the inflammatory cell infiltrate and cytokine production in wounds of equivalent size in oral mucosa and skin. Significantly lower levels of macrophage, neutrophil, and T-cell infiltration were observed in oral vs. dermal wounds. RT-PCR analysis of inflammatory cytokine production demonstrated that oral wounds contained significantly less IL-6 and KC than did skin wounds. Similarly, the level of the pro-fibrotic cytokine TGF-b1 was lower in mucosal than in skin wounds. No significant differences between skin and mucosal wounds were observed for the expression of the anti-inflammatory cytokine IL-10 and the TGF-beta1 modulators, fibromodulin and LTBP-1. These findings demonstrate that diminished inflammation is a key feature of the privileged repair of oral mucosa. PMID- 12885849 TI - Accelerated alveolar bone loss in mice lacking interleukin-10. AB - Interleukin-10 regulates pro-inflammatory cytokines, including those implicated in alveolar bone resorption. We hypothesized that lack of interleukin-10 leads to increased alveolar bone resorption. Male interleukin-10(-/-) mice, on 129/SvEv and C57BL/6J background, were compared with age-, sex-, and strain-matched interleukin-10(+/+) controls for alveolar bone loss. Immunoblotting was used for analysis of serum reactivity against bacteria associated with colitis and periodontitis. Interleukin-10(-/-) mice had significantly greater alveolar bone loss than interleukin-10(+/+) mice (p = 0.006). The 30-40% greater alveolar bone loss in interleukin-10(-/-) mice was evident in both strains, with C57BL/6J interleukin-10(-/-) mice exhibiting the most bone loss. Immunoblotting revealed distinct interleukin-10(-/-) serum reactivity against Bacteroides vulgatus, B. fragilis, Prevotella intermedia, and, to a lesser extent, against B. forsythus. The results of the present study suggest that lack of interleukin-10 leads to accelerated alveolar bone loss. PMID- 12885850 TI - Novel OK-432-conjugated tumor vaccines induce tumor-specific immunity against murine tongue cancer. AB - Priming with tumor antigens is one of the most important strategies in cancer immunotherapy. To enhance tumor antigenicity, OK-432, a streptococcal preparation, was coupled to squamous cell carcinoma (KLN-205) by means of a 0.2% glutaraldehyde method. The purpose of this study was to investigate whether OK 432-conjugated tumor vaccines could induce tumor-specific immunity. Our originally developed mouse tongue cancer model was used throughout this work for the analysis of antitumor effects. Prepared OK-432-conjugated KLN-205 vaccines were immunized 3 times to DBA/2 mice. The results showed that the KLN-205 vaccines induced cytolytic activity and strongly suppressed both KLN-205 tumor incidence and growth, and survival of the mice was improved. Moreover, the histological results showed that a greater number of lymphocytes had infiltrated around tumor cells by 24 hours after tumor inoculation in the vaccine group. These results suggest that immunizations with KLN-205 vaccines increase the antitumor effects against tongue cancer. PMID- 12885851 TI - Gene profiling in human periodontal ligament fibroblasts by subtractive hybridization. AB - Genes expressed by human periodontal ligament fibroblasts (HPFs) are likely to be associated with specific functions of the ligament. The aim of this study is to profile genes expressed highly by HPFs. A library (6 x 10(3) pfu) was constructed, followed by subtraction of HPF cDNAs with human gingival fibroblast (HGF) cDNAs. Reverse-dot hybridization revealed that 33 clones expressed higher levels of specific mRNAs in HPFs than in HGFs. These were mRNAs for known genes, including several associated with maturation and differentiation of cells. None had been reported in PFs. One clone, PDL-29, identified as a COX assembly factor, showed much stronger mRNA expression in HPFs than in HGFs in culture. In rat periodontium, however, PDL-29 mRNA expression was similar in PFs and GFs. These results suggest that HPFs express many previously unreported genes associated with maturation and differentiation, but expression can differ in vitro and in vivo. PMID- 12885852 TI - Expression of MMP-8 and MMP-13 genes in the periodontal ligament during tooth movement in rats. AB - Periodontal ligament tissue is remodeled on both the tension and compression sides of moving teeth during orthodontic tooth movement. The present study was designed to clarify the hypothesis that the expression of MMP-8 and MMP-13 mRNA is promoted during the remodeling of periodontal ligament tissue in orthodontic tooth movement. We used the in situ hybridization method and semi-quantitative reverse-transcription/polymerase chain-reaction analysis to elucidate the gene expression of MMP-8 and MMP-13 mRNA. Expression of MMP-8 and MMP-13 mRNA transiently increased on both the compression and tension sides during active tooth movement in vivo. The gene expression of MMP-8 and MMP-13 was induced by tension, while compression indirectly promoted the gene expression of MMP-8 and MMP-13 through soluble factors in vitro. Thus, we concluded that the expression of MMP-8 and MMP-13 is differentially regulated by tension and compression, and plays an important role in the remodeling of the periodontal ligament. PMID- 12885854 TI - Age-related changes in brain regional activity during chewing: a functional magnetic resonance imaging study. AB - Age-related changes in mastication-induced brain neuronal activity have been suggested. However, in humans, little is known about the anatomical regions involved. Using fMRI during cycles of rhythmic gum-chewing and no chewing, we have examined the effect of aging on brain regional activity during chewing in young adult (19-26 yrs), middle-aged (42-55 yrs), and aged (65-73 yrs) healthy humans. In all subjects, chewing resulted in a bilateral increase in the BOLD signals in the sensorimotor cortex, cerebellum, thalamus, supplementary motor area, and insula, and a unilateral increase in the right prefrontal area. In the first three regions, the signal increases were attenuated in an age-dependent manner, whereas, in the right prefrontal area, the converse was seen. The remaining two regions showed no significant differences with ages. These results indicate that chewing causes regional increases in neuronal activity in the brain, some of which are age-dependent. PMID- 12885853 TI - TGF-beta1 induces accumulation of dendritic cells in the odontoblast layer. AB - TGF-beta1 released from dentin degraded by bacterial or iatrogenic agents is suspected to influence dental pulp response, including the modulation of cell migration. To determine the consequences of TGF-beta1 action on pulp immune cells, we analyzed, by immunohistochemistry, the effect of transdentinally diffusing TGF-beta1 on their localization in a human tooth slice culture model. TGF-beta1 induced an accumulation of HLA-DR-positive cells in both odontoblast and subodontoblast layers of the stimulated zone. Together with HLA-DR, these cells co-expressed Factor XIIIa and CD68, two features of immature antigen presenting dendritic cells (DC), as well as the TGF-beta1 specific receptor TbetaRII. In contrast, no effect could be detected on the localization of either mature DC-LAMP-positive DC or of T- and B-lymphocytes. Analysis of these data suggests that TGF-beta1 released from dentin degraded by bacterial or iatrogenic agents could be involved in the immune response of the dental pulp resulting from tooth injury. PMID- 12885856 TI - Both xanthophyll cycle-dependent thermal dissipation and the antioxidant system are up-regulated in grape (Vitis labrusca L cv Concord) leaves in response to N limitation. AB - One-year-old grapevines (Vitis labrusca L. cv. Concord) were supplied with 0, 5, 10, 15, or 20 mM nitrogen (N) in a modified Hoagland's solution twice weekly for 4 weeks. As leaf N decreased in response to N limitation, leaf chlorophyll (Chl) decreased linearly whereas leaf absorptance declined curvilinearly. Compared with high N leaves, low N leaves had lower quantum efficiency of PSII as a result of both an increase in non-photochemical quenching (NPQ) and an increase in closure of PSII reaction centres at midday under high photon flux density (PFD). Both the xanthophyll cycle pool size on a Chl basis and the conversion of violaxanthin (V) to antheraxanthin (A) and zeaxanthin (Z) at noon increased with decreasing leaf N. NPQ was closely related to A+Z expressed either on a Chl basis or as a percentage of the xanthophyll cycle pool. As leaf N increased, superoxide dismutase (SOD) activity on a Chl basis decreased linearly; activities of catalase (CAT) and glutathione reductase (GR) on a Chl basis increased linearly; activities of ascorbate peroxidase (APX), monodehydroascorbate reductase (MDAR) and dehydroascorbate reductase (DHAR) expressed on the basis of Chl decreased rapidly first, then gradually reached a low level. In response to N limitation, the contents of ascorbate (AsA), dehydroascorbate (DAsA), reduced glutathione (GSH), and oxidized glutathione (GSSG) increased when expressed on a Chl basis, whereas the ratios of both AsA to DAsA and GSH to GSSG decreased. It is concluded that, in addition to decreasing light absorption by lowering Chl concentration, both xanthophyll cycle-dependent thermal energy dissipation and the antioxidant system are up-regulated to protect low N leaves from photo-oxidative damage under high light. PMID- 12885857 TI - Cavitation, stomatal conductance, and leaf dieback in seedlings of two co occurring Mediterranean shrubs during an intense drought. AB - Seedling shrubs in the Mediterranean semi-arid climate are subjected to intense droughts during summer. Thus, seedlings often surpass their limits of tolerance to water stress, resulting in the loss of hydraulic conductivity due to xylem cavitation. The response in terms of stomatal conductance, vulnerability to cavitation, leaf dieback, and survival were analysed in two co-occurring seedlings of mastic tree (Pistacia lentiscus L.) and kermes oak (Quercus coccifera L.) during an intense drought period. Both species reacted to drought with steep decreases in stomatal conductance before the critical water potential brought about the onset of cavitation events. Q. coccifera showed wider safety margins for avoiding runaway embolism than P. lentiscus and these differences could be related to the particular drought strategy displayed by each species: water saver or water spender. The limits for survival, resprout capacity and leaf dieback were also analysed in terms of loss of conductivity. By contrast with previous studies, the species showing higher seedling survival in the presence of drought also showed higher susceptibility to cavitation and operated with a lower safety margin for cavitation. Both species showed a leaf specific conductivity (LSC) threshold below which leaf biomass had to be regulated to avoid runaway embolism. However, each species displayed a different type of response: P. lentiscus conserved total leaf area up to 100% loss of LSC, whereas Q. coccifera continuously adjusted leaf biomass throughout the drought period in order to maintain the LSC very close to the maximum values recorded without loss of conductivity. Both species maintained the capacity for survival until the loss of conductivity was very nearly 100%. PMID- 12885858 TI - The nitrogen and nitrate economy of butterhead lettuce (Lactuca sativa var capitata L). AB - Quantifying and simulating the relationships between crop growth, total-nitrogen (total-N) and nitrate-N (NO3--N) concentration can improve crop nutritional husbandry. In this study, the relationship between shoot relative growth rate (RGR) and shoot total-N, organic-N and NO3--N concentration of hydroponically grown lettuce (Lactuca sativa var. capitata L. cv. Kennedy) was described and simulated. Plants were grown hydroponically for up to 74 d. Nitrogen was supplied throughout (control; T1), or removed at 35 d (T2) and 54 d (T3), respectively, after sowing. The organic-N and NO3--N concentration declined in the shoots of control plants with growth, until commercial maturity approached when organic-N and NO3--N concentration increased. There were sub-linear relationships between both total-N and organic-N concentration, and shoot RGR, in the N-limited treatments, i.e. shoot RGR approached an asymptote at high shoot N concentration. The proportional effects of total-N and organic-N concentration on shoot RGR were independent of plant age. A dynamic simulation model ('Nicolet'), derived previously under different conditions, was used to simulate the growth, dry matter content, organic-N, and NO3--N concentration of lettuce grown under the extreme N-stress conditions experienced by the plants. In view of the largely successful fitting of the model to experimental data, the model was used to interpret the results. Suggestions for model improvement are made. PMID- 12885859 TI - Relationships among shoot sinks for resources exported from nodal roots regulate branch development of distal non-rooted portions of Trifolium repens L. AB - Two manipulative experiments tested hypotheses pertaining to the correlative control exerted by nodal roots on branch development of the distal non-rooted portion of Trifolium repens growing clonally under near-optimal conditions. The two experiments, differing in their pattern of excision to manipulate the number of branches formed at the first 9-10 phytomers distal to the youngest nodal root, each found that after 20 phytomers of growth the total number of lateral branches formed on the primary stolon remained between five and seven regardless of where the branches formed along the stolon. Additional treatments established that nodal roots influenced branch development via relationships among shoot sinks for the root-supplied resources rather than through variation in the supply of such resources induced by fluctuations in photosynthate supply to roots from branches. Regression analysis of data pooled from treatments of both experiments confirmed that shoot-sink relationships for root- supplied resources controlled the branching processes on the non-rooted portion of plants. A disbudding treatment, which removed all the apical and axillary buds present on basal branches, but left other branch tissues intact, increased branch development of the apical region in the same way as did complete excision of the basal lateral branches. The apical buds and the elongation processes occurring immediately proximal to the buds were thus identified as strong sinks for the root-supplied resources. Such results suggest that branch development on the non-rooted shoot portion distal to the youngest nodal root is regulated by competition among sinks for root-derived resources, of limited availability, necessary for the processes of elongation of axillary buds and the primary stolon apical bud. PMID- 12885860 TI - Root cap removal increases root penetration resistance in maize (Zea mays L). AB - The root cap assists the passage of the root through soil by means of its slimy mucilage secretion and by the sloughing of its outer cells. The root penetration resistance of decapped primary roots of maize (Zea mays L. cv. Mephisto) was compared with that of intact roots in loose (dry bulk density 1.0 g cm-3; penetration resistance 0.06 MPa) and compact soil (1.4 g cm-3; penetration resistance 1.0 MPa), to evaluate the contribution of the cap to decreasing the impedance to root growth. Root elongation rate and diameter were the same for decapped and intact roots when the plants were grown in loose soil. In compacted soil, however, the elongation rate of decapped roots was only about half that of intact roots, whilst the diameter was 30% larger. Root penetration resistances of intact and decapped seminal axis were 0.31 and 0.52 MPa, respectively, when the roots were grown in compacted soil. These results indicated that the presence of a root cap alleviates much of the mechanical impedance to root penetration, and enables roots to grow faster in compacted soils. PMID- 12885861 TI - Alkaloids in plants and root cultures of Atropa belladonna overexpressing putrescine N-methyltransferase. AB - Putrescine N-methyltransferase (PMT) is the first alkaloid-specific enzyme for nicotine and tropane alkaloid formation. The pmt gene from Nicotiana tabacum was fused to the CaMV 35S promoter and integrated into the Atropa belladonna genome. Transgenic plants and derived root cultures were analysed for gene expression and for levels of alkaloids and their precursors. Scopolamine, hyoscyamine, tropine, pseudotropine, tropinone, and calystegines were found unaltered or somewhat decreased in pmt-overexpressing lines compared to controls. When root cultures were treated with 5% sucrose, calystegine levels were elevated in control roots, but were not affected in pmt-overexpressing roots. 1 microM auxin reduced calystegine levels in control roots, while in pmt-overexpressing roots all alkaloids remained unaltered. Expression level of pmt alone is apparently not limiting for tropane alkaloid formation in A. belladonna. PMID- 12885862 TI - Multiple forms of endo-1,4-beta-glucanases in the endosperm of Euphorbia heterophylla L. AB - Germinating seeds of Euphorbia heterophylla L. contain endo-1,4-beta-glucanases which degrade carboxymethylcellulose (CMC). The activity decreased approximately 66% in extracts of endosperm containing isopropanol or ethanol. The endoglucanases were isolated from endosperm extracts using ammonium sulphate fractionation followed by Sephacryl S-100-HR chromatography resulting in two main peaks: I and II. Peak I endoglucanase was further purified about 15-fold on DEAE Sephadex A50 and then by affinity chromatography (CF11-cellulose). Peak II endoglucanases were further purified 10-fold on CM-cellulose chromatography. The results indicated the occurrence of a 66 kDa endoglucanase (fractionated by SDS PAGE and visualized by activity staining using Congo Red). Several acidic (pI 3.0 to 5.7) and basic (pI 8.5 to 10.0) forms from both peaks which differed in their capacities for degrading CMC or xyloglucans from Copaifera langsdorffii or Hymenaea courbaril were detected. PMID- 12885863 TI - Identification and localization of a beta-COP-like protein involved in the morphodynamics of the plant Golgi apparatus. AB - This paper examines the molecular machinery involved in membrane exchange within the plant endomembrane system. A study has been undertaken on beta-COP-like proteins in plant cells using M3A5, an antibody raised against the conserved sequence of mammalian beta-COP proteins. In mammalian cells, beta-COP proteins are part of a complex named the coatomer, which probably recruits some specific areas of the endomembrane system. Immunofluorescence analyses by confocal laser scanning microscopy showed that beta-COP-like proteins marked predominantly the plant Golgi apparatus. Other proteins known to be part of a potential machinery for COPI vesicle formation (gamma-COP, beta'-COP and Arf1 proteins) were immunolocalized on the same membraneous structures as beta-COP. Moreover, beta COP and other COPI antibodies stained the cell plate in dividing cells. It is further shown that, in maize root cells, and in contrast to observations upon mammalian cells, the drug Brefeldin A (BFA) does not induce the release of beta COP and Arf1 proteins from the Golgi membrane into the cytosol. These data clearly demonstrate that the antibody M3A5 is a valuable marker for studies on trafficking events in plant cells. They also report for the first time the location of COP components in plant tissue at the light level, especially on a model well known for secretion, i.e. the maize root cells. They also suggest that the membrane recruitment machinery may function in a plant-specific way. PMID- 12885864 TI - Expression analysis of a GA 20-oxidase in embryos from two sorghum lines with contrasting dormancy: possible participation of this gene in the hormonal control of germination. AB - The role of GAs in promoting seed germination is well known and experiments with seeds from different species have suggested the requirement of de novo synthesis of GAs upon imbibition for germination. There are also strong indications that the enhancement of GA synthesis is part of the mechanism through which environmental signals (i.e. light) induce germination. Since along the GA biosynthetic pathway, oxidation at C-20 carried out by GA 20-oxidases is thought to be a site of regulation, a cDNA clone encoding a GA 20-oxidase was isolated from embryos of sorghum (SbGA 20ox). Expression analysis of this gene in embryos within imbibed caryopses with low dormancy showed detectable amounts of the specific mRNA early upon incubation, increasing thereafter. In contrast, it remained barely detectable in embryos from dormant caryopses. Changes in endogenous GA4 levels were in agreement with those of SbGA 20ox mRNA, suggesting that GA production might be regulated differentially at the level of transcription of this gene. The expression of SbGA 20ox was enhanced in incubated embryos isolated from either type of caryopses, illustrating a physiological control exerted by the surrounding seed tissues on gene expression. The results also show that ABA leads to a suppression of transcription of this gene. PMID- 12885865 TI - Cloning of a sucrose-phosphate synthase gene highly expressed in flowers from the tropical epiphytic orchid Oncidium Goldiana. AB - Sucrose-phosphate synthase (SPS) is one of the key regulatory enzymes in carbon assimilation and partitioning in plants. It plays a crucial role in the production of sucrose in photosynthetic cells. The cloning and expression analysis of a full-length cDNA encoding SPS from tropical epiphytic orchid hybrid Oncidium Goldiana are reported here. The cDNA designated as sps1 is 3820 bp in length with an open reading frame of 3183 bp encoding 1061 amino acids. The deduced amino acid sequence of O. Goldiana sps1 shows 56% and 69% homology with those of maize SPS and spinach SPS, respectively. The high level expression of O. Goldiana sps1 in the flower suggests that it might play an important role in flowering. Growth under higher irradiance and elevated CO2 leads to an accumulation of the sps1 transcript in the photosynthetic leaves. It appears that SPS gene expression in photosynthetic leaves is associated with the leaf photosynthetic rate. PMID- 12885866 TI - Vessel contents of leaves after excision: a test of the Scholander assumption. AB - When petioles of transpiring leaves are cut in the air, according to the 'Scholander assumption', the vessels cut open should fill with air as the water is drained away by tissue rehydration and/or continued transpiration. The distribution of air-filled vessels versus distance from the cut surface should match the distribution of lengths of 'open vessels', i.e. vessels cut open when the leaf is excised. A paint perfusion method was used to estimate the length distribution of open vessels and this was compared with the observed distribution of embolisms by the cryo-SEM method. In the cryo-SEM method, petioles are frozen in liquid nitrogen soon after the petiole is cut. The petioles are then cut at different distances from the original cut surface while frozen and examined in a cryo-SEM facility, where it is easy to distinguish vessels filled with air from those filled with ice. The Scholander assumption was also confirmed by a hydraulic method, which avoided possible freezing artefacts. In petioles of sunflower (Helianthus annuus L) the distribution of embolized vessels agrees with expectations. This is in contrast to a previous study on sunflower where cryo-SEM results did not agree with expectations. The reasons for this disagreement are suggested, but further study is required for a full elucidation. PMID- 12885867 TI - Putative PIP1 genes isolated from apple: expression analyses during fruit development and under osmotic stress. AB - To gain insight into the function of plasma membrane intrinsic protein (PIP) genes in apple, two genes, MdPIP1a and MdPIP1b, were isolated. MdPIP1 expression was in accordance with the volume increase during fruit development, which is a loading process of water and solutes. In addition, the expression of MdPIP1 was up-regulated in the stems by osmotic stress. These results indicate that MdPIP1 may play important roles not only in fruit expansion, but also in maintaining water homeostasis under stress conditions. PMID- 12885868 TI - P-selectin and P-selectin glycoprotein ligand 1 are major determinants for Th1 cell recruitment to nonlymphoid effector sites in the intestinal lamina propria. AB - The recruitment of activated T cell subsets to sites of effector immune responses is mediated by homing receptors induced upon activation in secondary lymphoid tissue. Using an adoptive transfer model, the intestinal recruitment of CD4+ T cells activated with intraperitoneal antigen in complete Freund's adjuvant was examined. The data demonstrate that activated CD4+ T cells recruited to intestinal Peyer's patches (PP) and lamina propria (LP) up-regulate functional P selectin glycoprotein ligand 1 (PSGL-1). Blockade of IL-12 inhibited functional PSGL-1 expression and reduced PP and LP CD4+ T cell recruitment by >40%. P selectin blockade reduced LP recruitment of activated cells by 56% without affecting PP recruitment. Studies of mice examined 3 d after adoptive transfer of differentiated T cell subsets revealed that Th1 but not Th2 cells were recruited to small intestine PP and LP. Mucosal addressin cell adhesion molecule blockade reduced Th1 recruitment to PP by 90% and to LP by >72%, whereas P-selectin blockade reduced Th1 recruitment to PP by 18% and Th1 recruitment to LP by 84%. These data suggest that IL-12-induced functional PSGL-1 expression is a major determinant for the recruitment of Th1 effector cells to noninflamed as well as inflamed intestine. PMID- 12885869 TI - A monoclonal antibody to the alpha2 domain of murine major histocompatibility complex class I that specifically kills activated lymphocytes and blocks liver damage in the concanavalin A hepatitis model. AB - We earlier found that a rat monoclonal antibody (mAb) RE2 can induce rapid death of murine activated, but not resting, lymphocytes and lymphocyte cell lines, in a complement-independent manner, a cell death differing from typical apoptosis or necrosis. We here found that this cell death is independent of pathways involving Fas, caspase, and phosphoinositide-3 kinase. With the advantage of producing human B cell line transfectants with stable expression of human/mouse xeno chimeric MHC class I genes, we found that RE2 epitope resides on the murine class I alpha2 domain. However, the alpha3 domain plays a key role in transducing the death signal, which mediates extensive aggregation of the MHC class I-integrin actin filament system, giving rise to membrane blebs and pores. In mouse models with T/NKT cell activation-associated fulminant hepatitis, administration of mAb RE2 almost completely inhibited the development of liver cell injuries. Taken collectively, this form of cell death may be involved in homeostatic immune regulation, and induction of this form of cell death using the mAbs may be potentially therapeutic for subjects with immunological diseases mediated by activated lymphocytes. PMID- 12885870 TI - Vav1 phosphorylation is induced by beta2 integrin engagement on natural killer cells upstream of actin cytoskeleton and lipid raft reorganization. AB - The guanine nucleotide exchange factor Vav1 regulates actin polymerization and contributes to cytotoxicity by natural killer (NK) cells. An open question is how Vav1 becomes activated and what receptor can signal upstream of actin cytoskeleton rearrangement upon NK cell contact with target cells. Using transfected insect cells that express ligands of human NK cell receptors, we show that engagement of the beta2 integrin LFA-1 on NK cells by intercellular adhesion molecule (ICAM)-1 led to a tyrosine phosphorylation of Vav1 that was not sensitive to cholesterol depletion and to inhibition of actin polymerization. Vav1 phosphorylation was blocked by an inhibitor of Src-family kinases, and correlated with activation of its downstream effector PAK. Binding of activation receptor 2B4 to its ligand CD48 was not sufficient for Vav1 phosphorylation. However, coengagement of 2B4 with LFA-1 resulted in an enhancement of Vav1 phosphorylation that was sensitive to cholesterol depletion and to inhibition of actin polymerization. Vav1 was recruited to a detergent-resistant membrane (DRM) fraction only when 2B4 and LFA-1 were coengaged, but not after LFA-1 engagement. Therefore, binding of LFA-1 to ICAM-1 on target cells may initiate an early signaling cascade in NK cells through activation of Vav1, leading to cytoskeleton reorganization and amplification of signals from other activation receptors. PMID- 12885871 TI - Differential antigen presentation regulates the changing patterns of CD8+ T cell immunodominance in primary and secondary influenza virus infections. AB - The specificity of CD8+ T cell responses can vary dramatically between primary and secondary infections. For example, NP366-374/Db- and PA224-233/Db-specific CD8+ T cells respond in approximately equal numbers to a primary influenza virus infection in C57BL/6 mice, whereas NP366-374/Db-specific CD8+ T cells dominate the secondary response. To investigate the mechanisms underlying this changing pattern of immunodominance, we analyzed the role of antigen presentation in regulating the specificity of the T cell response. The data show that both dendritic and nondendritic cells are able to present the NP366-374/Db epitope, whereas only dendritic cells effectively present the PA224-233/Db epitope after influenza virus infection, both in vitro and in vivo. This difference in epitope expression favored the activation and expansion of NP366-374/Db-specific CD8+ memory T cells during secondary infection. The data also show that the immune response to influenza virus infection may involve T cells specific for epitopes, such as PA224-233/Db, that are poorly expressed at the site of infection. In this regard, vaccination with the PA224-233 peptide actually had a detrimental effect on the clearance of a subsequent influenza virus infection. Thus, differential antigen presentation impacts both the specificity of the T cell response and the efficacy of peptide-based vaccination strategies. PMID- 12885872 TI - Regulation of cyclooxygenase-2 expression by the translational silencer TIA-1. AB - The cyclooxygenase-2 (COX-2) enzyme catalyzes the rate-limiting step of prostaglandin formation in inflammatory states, and COX-2 overexpression plays a key role in carcinogenesis. To understand the mechanisms regulating COX-2 expression, we examined its posttranscriptional regulation mediated through the AU-rich element (ARE) within the COX-2 mRNA 3'-untranslated region (3'UTR). RNA binding studies, performed to identify ARE-binding regulatory factors, demonstrated binding of the translational repressor protein TIA-1 to COX-2 mRNA. The significance of TIA-1-mediated regulation of COX-2 expression was observed in TIA-1 null fibroblasts that produced significantly more COX-2 protein than wild type fibroblasts. However, TIA-1 deficiency did not alter COX-2 transcription or mRNA turnover. Colon cancer cells demonstrated to overexpress COX-2 through increased polysome association with COX-2 mRNA also showed defective TIA-1 binding both in vitro and in vivo. These findings implicate that TIA-1 functions as a translational silencer of COX-2 expression and support the hypothesis that dysregulated RNA-binding of TIA-1 promotes COX-2 expression in neoplasia. PMID- 12885873 TI - Multiple T cell subsets control Francisella tularensis LVS intracellular growth without stimulation through macrophage interferon gamma receptors. AB - A variety of data suggest that in vivo production of interferon (IFN)-gamma is necessary, but not sufficient, for expression of secondary protective immunity against intracellular pathogens. To discover specific IFN-gamma-independent T cell mediated mechanisms, we took advantage of an in vitro culture system that models in vivo immune responses to the intracellular bacterium Francisella tularensis live vaccine strain (LVS). LVS-immune lymphocytes specifically controlled 99% of the growth of LVS in wild-type murine bone marrow-derived macrophages. Surprisingly, LVS-immune lymphocytes also inhibited LVS intracellular growth by as much as 95% in macrophages derived from IFN-gamma receptor knockout (IFNgammaR KO) mice. CD8+ T cells, and to a lesser degree CD4+ T cells, controlled LVS intracellular growth in both wild-type and IFNgammaR KO macrophages. Further, a unique population of Thy1+alphabeta+CD4-CD8- cells that was previously suggested to operate during secondary immunity to LVS in vivo strongly controlled LVS intracellular growth in vitro. A large proportion of the inhibition of LVS intracellular growth in IFNgammaR KO macrophages by all three T cell subsets could be attributed to tumor necrosis factor (TNF) alpha. Thus, T cell mechanisms exist that control LVS intracellular growth without acting through the IFN-gamma receptor; such control is due in large part to TNF-alpha, and is partially mediated by a unique double negative T cell subpopulation. PMID- 12885874 TI - ClC channels: reading eukaryotic function through prokaryotic spectacles. PMID- 12885875 TI - Side-chain charge effects and conductance determinants in the pore of ClC-0 chloride channels. AB - The charge on the side chain of the internal pore residue lysine 519 (K519) of the Torpedo ClC-0 chloride (Cl-) channel affects channel conductance. Experiments that replace wild-type (WT) lysine with neutral or negatively charged residues or that modify the K519C mutant with various methane thiosulfonate (MTS) reagents show that the conductance of the channel decreases when the charge at position 519 is made more negative. This charge effect on the channel conductance diminishes in the presence of a high intracellular Cl- concentration ([Cl-]i). However, the application of high concentrations of nonpermeant ions, such as glutamate or sulfate (SO42-), does not change the conductance, suggesting that the electrostatic effects created by the charge at position 519 are unlikely due to a surface charge mechanism. Another pore residue, glutamate 127 (E127), plays an even more critical role in controlling channel conductance. This negatively charged residue, based on the structures of the homologous bacterial ClC channels, lies 4-5 A from K519. Altering the charge of this residue can influence the apparent Cl- affinity as well as the saturated pore conductance in the conductance-Cl- activity curve. Amino acid residues at the selectivity filter also control the pore conductance but mutating these residues mainly affects the maximal pore conductance. These results suggest at least two different conductance determinants in the pore of ClC-0, consistent with the most recent crystal structure of the bacterial ClC channel solved to 2.5 A, in which multiple Cl--binding sites were identified in the pore. Thus, we suggest that the occupancy of the internal Cl--binding site is directly controlled by the charged residues located at the inner pore mouth. On the other hand, the Cl--binding site at the selectivity filter controls the exit rate of Cl- and therefore determines the maximal channel conductance. PMID- 12885876 TI - Probing the pore of ClC-0 by substituted cysteine accessibility method using methane thiosulfonate reagents. AB - ClC channels are a family of protein molecules containing two ion-permeation pores. Although these transmembrane proteins are important for a variety of physiological functions, their molecular operations are only superficially understood. High-resolution X-ray crystallography techniques have recently revealed the structures of two bacterial ClC channels, but whether vertebrate ClC channel pores are similar to those of bacterial homologues is not clear. To study the pore architecture of the Torpedo ClC-0 channel, we employed the substituted cysteine-accessibility method (SCAM) and used charged methane thiosulfonate (MTS) compounds to modify the introduced cysteine. Several conclusions were derived from this approach. First, the MTS modification pattern from Y512C to E526C in ClC-0, which corresponds to residues forming helix R in bacterial ClC channels, is indeed consistent with the suggested helical structure. Second, the ClC-0 pore is more accessible to the negatively charged than to the positively charged MTS compound, a pore property that is regulated by the intrinsic electrostatic potential in the pore. Finally, attempts to modify the introduced cysteine at positions intracellular to the selectivity filter did not result in larger MTS modification rates for the open-state channel, suggesting that the fast gate of ClC-0 cannot be located at a position intracellular to the Cl- selectivity filter. Thus, the proposal that the glutamate side chain is the fast gate of the channel is applicable to ClC-0, revealing a structural and functional conservation of ClC channels between bacterial and vertebrate species. PMID- 12885877 TI - Stabilization of the activity of ATP-sensitive potassium channels by ion pairs formed between adjacent Kir6.2 subunits. AB - ATP-sensitive potassium (KATP) channels are formed by the coassembly of four Kir6.2 subunits and four sulfonylurea receptor subunits (SUR). The cytoplasmic domains of Kir6.2 mediate channel gating by ATP, which closes the channel, and membrane phosphoinositides, which stabilize the open channel. Little is known, however, about the tertiary or quaternary structures of the domains that are responsible for these interactions. Here, we report that an ion pair between glutamate 229 and arginine 314 in the intracellular COOH terminus of Kir6.2 is critical for maintaining channel activity. Mutation of either residue to alanine induces inactivation, whereas charge reversal at positions 229 and 314 (E229R/R314E) abolishes inactivation and restores the wild-type channel phenotype. The close proximity of these two residues is demonstrated by disulfide bond formation between cysteine residues introduced at the two positions (E229C/R314C); disulfide bond formation abolishes inactivation and stabilizes the current. Using Kir6.2 tandem dimer constructs, we provide evidence that the ion pair likely forms by residues from two adjacent Kir6.2 subunits. We propose that the E229/R314 intersubunit ion pairs may contribute to a structural framework that facilitates the ability of other positively charged residues to interact with membrane phosphoinositides. Glutamate and arginine residues are found at homologous positions in many inward rectifier subunits, including the G-protein activated inwardly rectifying potassium channel (GIRK), whose cytoplasmic domain structure has recently been solved. In the GIRK structure, the E229- and R314 corresponding residues are oriented in opposite directions in a single subunit such that in the tetramer model, the E229 equivalent residue from one subunit is in close proximity of the R314 equivalent residue from the adjacent subunit. The structure lends support to our findings in Kir6.2, and raises the possibility that a homologous ion pair may be involved in the gating of GIRKs. PMID- 12885878 TI - External TEA block of shaker K+ channels is coupled to the movement of K+ ions within the selectivity filter. AB - Recent molecular dynamic simulations and electrostatic calculations suggested that the external TEA binding site in K+ channels is outside the membrane electric field. However, it has been known for some time that external TEA block of Shaker K+ channels is voltage dependent. To reconcile these two results, we reexamined the voltage dependence of block of Shaker K+ channels by external TEA. We found that the voltage dependence of TEA block all but disappeared in solutions in which K+ ions were replaced by Rb+. These and other results with various concentrations of internal K+ and Rb+ ions suggest that the external TEA binding site is not within the membrane electric field and that the voltage dependence of TEA block in K+ solutions arises through a coupling with the movement of K+ ions through part of the membrane electric field. Our results suggest that external TEA block is coupled to two opposing voltage-dependent movements of K+ ions in the pore: (a) an inward shift of the average position of ions in the selectivity filter equivalent to a single ion moving approximately 37% into the pore from the external surface; and (b) a movement of internal K+ ions into a vestibule binding site located approximately 13% into the membrane electric field measured from the internal surface. The minimal voltage dependence of external TEA block in Rb+ solutions results from a minimal occupancy of the vestibule site by Rb+ ions and because the energy profile of the selectivity filter favors a more inward distribution of Rb+ occupancy. PMID- 12885879 TI - Potent, persistent induction and modulation of cellular immune responses in rhesus macaques primed with Ad5hr-simian immunodeficiency virus (SIV) env/rev, gag, and/or nef vaccines and boosted with SIV gp120. AB - Immunity elicited by multicomponent vaccines delivered by replication-competent Ad5hr-simian immunodeficiency virus (SIV) recombinants was systematically investigated. Rhesus macaques were immunized mucosally at weeks 0 and 12 with Ad5hr-SIV(smH4) env/rev, with or without Ad5hr-SIV(mac239) gag or Ad5hr SIV(mac239) nef, or with all three recombinants. The total Ad5hr dosage was comparably adjusted among all animals with empty Ad5hr-DeltaE3 vector. The macaques were boosted with SIV gp120 in monophosphoryl A-stable emulsion adjuvant at 24 and 36 weeks. Controls received Ad5hr-DeltaE3 vector or adjuvant only. By ELISPOT analysis, all four SIV gene products elicited potent cellular immune responses that persisted 42 weeks post-initial immunization. Unexpectedly, modulation of this cellular immune response was observed among macaques receiving one, two, or three Ad5hr-SIV recombinants. Env responses were significantly enhanced throughout the immunization period in macaques immunized with Ad5hr-SIV env/rev plus Ad5hr-SIV gag and tended to be higher in macaques that also received Ad5hr-SIV nef. Macaques primed with all three recombinants displayed significant down-modulation in numbers of gamma interferon (IFN-gamma)-secreting cells specific for SIV Nef, and the Env- and Gag-specific responses were also diminished. Modulation of antibody responses was not observed. Down-modulation was seen only during the period of Ad5hr-recombinant priming, not during subunit boosting, although SIV-specific IFN-gamma-secreting cells persisted. The effect was not attributable to Ad5hr replication differences among immunization groups. Vaccine delivery via replication-competent live vectors, which can persistently infect new cells and continuously present low-level antigen, may be advantageous in overcoming competition among complex immunogens for immune recognition. Effects of current multicomponent vaccines on individual immune responses should be evaluated with regard to future vaccine design. PMID- 12885880 TI - Diminished RNA primer usage associated with the L74V and M184V mutations in the reverse transcriptase of human immunodeficiency virus type 1 provides a possible mechanism for diminished viral replication capacity. AB - The emergence of drug resistance-conferring mutations can severely compromise the success of chemotherapy directed against human immunodeficiency virus type 1 (HIV 1). The M184V and/or L74V mutation in the reverse transcriptase (RT) gene are frequently found in viral isolates from patients treated with the nucleoside RT inhibitors lamivudine (3TC), abacavir (ABC), and didanosine (ddI). However, the effectiveness of combination therapy with regimens containing these compounds is often not abolished in the presence of these mutations; it has been conjectured that diminished fitness of HIV-1 variants containing L74V and M184V may contribute to sustained antiviral effects in such cases. We have determined that viruses containing both L74V and M184V are more impaired in replication capacity than viruses containing either mutation alone. To understand the biochemical mechanisms responsible for this diminished fitness, we generated a series of recombinant mutated enzymes containing either or both of the L74V and M184V substitutions. These enzymes were tested for their abilities to bypass important rate-limiting steps during the complex process of reverse transcription. We studied both the initiation of minus-strand DNA synthesis with the cognate replication primer human tRNA(3)(Lys) and the initiation of plus-strand DNA synthesis, using a short RNA primer derived from the viral polypurine tract. We observed that the efficiencies of both reactions were diminished with enzymes containing either L74V or M184V and that these effects were significantly amplified with the double mutant. We also show that release from intrinsic pausing sites during reverse transcription appears to be a major obstacle that cannot be efficiently bypassed. Our data suggest that the efficiency of RNA primed DNA synthesis represents an important consideration that can affect viral replication kinetics. PMID- 12885881 TI - Identification of foot-and-mouth disease virus-specific linear B-cell epitopes to differentiate between infected and vaccinated cattle. AB - Foot-and-mouth disease (FMD) is a highly contagious viral disease of cloven hoofed animals. For several years, vaccination of animals, which had proven to be successful for the eradication of the disease, has been forbidden in the United States and the European Community because of the difficulty of differentiating between vaccinated and infected animals. In this study, detailed investigations of the bovine humoral immune response against FMD virus (FMDV) were performed with the aim of identifying viral epitopes recognized specifically by sera derived from FMDV-infected animals. The use of overlapping 15-mer synthetic peptides, covering the whole open reading frame of FMDV strain O(1)K in a peptide enzyme-linked immunosorbent assay, allowed the identification of 12 FMDV strain O(1)K-specific linear B-cell epitopes. Six of these linear B-cell epitopes, located in the nonstructural proteins, were used in further assays to compare the reactivities of sera from vaccinated and infected cattle. Antibodies recognizing these peptides could be detected only in sera derived from infected cattle. In further experiments, the reactivity of the six peptides with sera from animals infected with different strains of FMDV was tested, and strain-independent infection-specific epitopes were identified. Thus, these results clearly demonstrate the ability of a simple peptide-based assay to discriminate between infected and conventionally FMD-vaccinated animals. PMID- 12885882 TI - Crimean-Congo hemorrhagic fever virus glycoprotein proteolytic processing by subtilase SKI-1. AB - Crimean-Congo hemorrhagic fever (CCHF) virus is a tick-borne member of the genus Nairovirus, family Bunyaviridae. The mature virus glycoproteins, Gn and Gc (previously referred to as G2 and G1), are generated by proteolytic cleavage from precursor proteins. The amino termini of Gn and Gc are immediately preceded by tetrapeptides RRLL and RKPL, respectively, leading to the hypothesis that SKI-1 or related proteases may be involved (A. J. Sanchez, M. J. Vincent, and S. T. Nichol, J. Virol. 76:7263-7275, 2002). In vitro peptide cleavage data show that an RRLL peptide representing the Gn processing site is efficiently cleaved by SKI 1 protease, whereas an RKPL peptide representing the Gc processing site is cleaved at negligible levels. The efficient cleavage of RRLL peptide is consistent with the known recognition sequences of SKI-1, including the sequence determinants involved in the cleavage of the Lassa virus (family Arenaviridae) glycoprotein precursor. These in vitro findings were confirmed by expression of wild-type or mutant CCHF virus glycoproteins in CHO cells engineered to express functional or nonfunctional SKI-1. Gn processing was found to be dependent on functional SKI-1, whereas Gc processing was not. Gn processing occurred in the endoplasmic reticulum-cis Golgi compartments and was dependent on an R at the -4 position within the RRLL recognition motif, consistent with the known cleavage properties of SKI-1. Comparison of SKI-1 cleavage efficiency between peptides representing Lassa virus GP2 and CCHF virus Gn cleavage sites suggests that amino acids flanking the RRLL may modulate the efficiency. The apparent lack of SKI-1 cleavage at the CCHF virus Gc RKPL site indicates that related proteases, other than SKI-1, are likely to be involved in the processing at this site and identical or similar sites utilized in several New World arenaviruses. PMID- 12885883 TI - Reverse genetic system for the analysis of parvovirus telomeres reveals interactions between transcription factor binding sites in the hairpin stem. AB - The left-hand or 3'-terminal hairpin of minute virus of mice (MVM) contains sequence elements essential for both viral DNA replication at the left-hand origin (oriL) and for the modulation of the P4 promoter, from which the viral nonstructural gene cassette is transcribed. This hairpin sequence has proven difficult to manipulate in the context of the viral genome. Here we describe a system for generating mutant viruses using synthetic hairpin oligonucleotides and a truncated form of the infectious clone. This allows manipulation of the sequence of the left-hand hairpin and examination of the effects in the context of the viral life cycle. We have confirmed the requirement for a functional parvovirus initiation factor (PIF) binding site and determined that an optimized PIF binding site, with 6 bases between the half-sites, was actually detrimental to viral growth. The distal PIF half-site overlaps a cyclic AMP-responsive element (CRE), which was shown to play an important role in initiating infection, particularly in 324K simian virus 40-transformed human fibroblasts. Interestingly, reducing the spacing of the PIF half-sites, and thus the affinity of the binding site for PIF, increased viral fitness relative to wild type in 324K cells, but not in murine A9 cells. These results indicate that the relative importance of factor binding to the CRE and PIF sites during the establishment of an infection differs markedly between these two host cells and suggest that the suboptimal spacing of PIF half-sites found in wild-type virus represents a necessary reduction in the affinity of the PIF interaction in favor of CRE function. PMID- 12885884 TI - Nonstructural proteins NS1 and NS2 of bovine respiratory syncytial virus block activation of interferon regulatory factor 3. AB - We have previously shown that the nonstructural (NS) proteins NS1 and NS2 of bovine respiratory syncytial virus (BRSV) mediate resistance to the alpha/beta interferon (IFN)-mediated antiviral response. Here, we show that they, in addition, are able to prevent the induction of beta IFN (IFN-beta) after virus infection or double-stranded RNA stimulation. In BRSV-infected MDBK cells upregulation of IFN-stimulated genes (ISGs) such as MxA did not occur, although IFN signaling via JAK/STAT was found intact. In contrast, infection with recombinant BRSVs lacking either or both NS genes resulted in efficient upregulation of ISGs. Biological IFN activity and IFN-beta were detected only in supernatants of cells infected with the NS deletion mutants but not with wild type (wt) BRSV. Subsequent analyses of IFN-beta promoter activity showed that infection of cells with the double deletion mutant BRSV DeltaNS1/2, but not with BRSV wt, resulted in a significant increase in IFN-beta gene promoter activity. Induction of the IFN-beta promoter depends on the activation of three distinct transcription factors, NF-kappaB, ATF-2/c-Jun, and IFN regulatory factor 3 (IRF 3). Whereas NF-kappaB and ATF-2/c-Jun activities were readily detectable and comparable in both wt BRSV- and BRSV DeltaNS1/2-infected cells, phosphorylation and transcriptional activity of IRF-3, however, were observed only after BRSV DeltaNS1/2 infection. NS protein-mediated inhibition of IRF-3 activation and IFN induction should have considerable impact on the pathogenesis and immunogenicity of BRSV. PMID- 12885885 TI - Identification of a region of the tobacco mosaic virus 126- and 183-kilodalton replication proteins which binds specifically to the viral 3'-terminal tRNA-like structure. AB - UV irradiation of a mixture of an isolated tobacco mosaic virus (TMV; tomato strain L [TMV-L]) RNA-dependent RNA polymerase complex and the TMV-L RNA 3' terminal region (3'-TR) resulted in cross-linking of the TMV-L 126-kDa replication protein to the TMV-L 3'-TR. Using both Escherichia coli-expressed proteins corresponding to parts of the 126-kDa protein and mutants of the 3'-TR, the interacting sites were located to a 110-amino-acid region just downstream of the core methyltransferase domain in the protein and a region comprising the central core C and domain D2 in the 3'-TR. Mutation to alanine of a tyrosine residue at position 409 or a tyrosine residue at position 416 in the protein binding region abolished cross-linking to the 3'-TR, and corresponding mutations introduced into TMV-L RNA abolished its ability to replicate in tomato protoplasts, with no detectable production of either plus- or minus-strand RNA. The results are compatible with a model for initiation of TMV-L minus-strand RNA synthesis in which an internal region of the TMV-L 126-kDa protein first binds to the central core C and domain D2 region of the TMV-L 3'-TR and is then followed by binding of the 183-kDa protein to this complex and positioning of the catalytically active site of the polymerase domain close to the 3'-terminal CCCA initiation site. PMID- 12885886 TI - Newcastle disease virus V protein is associated with viral pathogenesis and functions as an alpha interferon antagonist. AB - Newcastle disease virus (NDV) edits its P gene by inserting one or two G residues at the conserved editing site (UUUUUCCC, genome sense) and transcribes the P mRNA (unedited), the V mRNA (with a +1 frameshift), and the W mRNA (with a +2 frameshift). All three proteins are amino coterminal but vary at their carboxyl terminus in length and amino acid composition. Little is known about the role of the V and W proteins in NDV replication and pathogenesis. We have constructed and recovered two recombinant viruses in which the expression of the V or both the V and W proteins has been abolished. Compared to the parental virus, the mutant viruses showed impaired growth in cell cultures, except in Vero cells. However, transient expression of the carboxyl-terminal portion of the V protein enhanced the growth of the mutant viruses. In embryonated chicken eggs, the parental virus grew to high titers in embryos of different gestational ages, whereas the mutant viruses showed an age-dependent phenomenon, growing to lower titer in more developed embryos. An interferon (IFN) sensitivity assay showed that the parental virus was more resistant to the antiviral effect of IFN than the mutant viruses. Moreover, infection with the parental virus resulted in STAT1 protein degradation, but not with the mutant viruses. These findings indicate that the V protein of NDV possesses the ability to inhibit alpha IFN and that the IFN inhibitory function lies in the carboxyl-terminal domain. Pathogenicity studies showed that the V protein of NDV significantly contributes to the virus virulence. PMID- 12885887 TI - PML residue lysine 160 is required for the degradation of PML induced by herpes simplex virus type 1 regulatory protein ICP0. AB - During the early stages of herpes simplex virus type 1 (HSV-1) infection, viral immediate-early regulatory protein ICP0 localizes to and disrupts cellular nuclear structures known as PML nuclear bodies or ND10. These activities correlate with the functions of ICP0 in stimulating lytic infection and reactivating quiescent HSV-1. The disruption of ND10 occurs because ICP0 induces the loss of the SUMO-1-modified forms of PML and the subsequent proteasome mediated degradation of the PML protein. The functions of ICP0 are largely dependent on the integrity of its zinc-binding RING finger domain. Many RING finger proteins have been found to act as ubiquitin E3 ligase enzymes, stimulating the production of conjugated polyubiquitin chains in the presence of ubiquitin, the ubiquitin-activating enzyme E1, and the appropriate E2 ubiquitin conjugating enzyme. Substrate proteins that become polyubiquitinated are then subject to degradation by proteasomes. We have previously shown that purified full-length ICP0 acts as an efficient E3 ligase in vitro, producing high molecular-weight polyubiquitin chains in a RING finger-dependent but substrate independent manner. In this paper we report on investigations into the factors governing the degradation of PML induced by ICP0 in a variety of in vivo and in vitro assays. We found that ICP0 expression increases the levels of ubiquitinated PML in transfected cells. However, ICP0 does not interact with or directly ubiquitinate either unmodified PML or SUMO-1-modified PML in vitro, suggesting either that additional factors are required for the ICP0-mediated ubiquitination of PML in vivo or that PML degradation is an indirect consequence of some other activity of ICP0 at ND10. Using a transfection-based approach and a family of deletion and point mutations of PML, we found that efficient ICP0-induced PML degradation requires sequences within the C-terminal part of PML and lysine residue 160, one of the principal targets for SUMO-1 modification of the protein. PMID- 12885888 TI - Selection of retroviral reverse transcription primer is coordinated with tRNA biogenesis. AB - Initiation of retrovirus reverse transcription requires the selection of a tRNA primer from the intracellular milieu. To investigate the features of primer selection, a human immunodeficiency virus type 1 (HIV-1) and a murine leukemia virus (MuLV) were created that require yeast tRNA(Phe) to be supplied in trans for infectivity. Wild-type yeast tRNA(Phe) expressed in mammalian cells was transported to the cytoplasm and aminoacylated. In contrast, tRNA(Phe) without the D loop (tRNA(Phe)D(-)) was retained within the nucleus and did not complement infectivity of either HIV-1 or MuLV; however, infectivity was restored when tRNA(Phe)D(-) was directly transfected into the cytoplasm of cells. A tRNA(Phe) mutant (tRNA(Phe)UUA) that did not have the capacity to be aminoacylated was transported to the cytoplasm and did complement infectivity of both HIV-1 and MuLV, albeit at a level less than that with wild-type tRNA(Phe). Collectively, our results demonstrate that the tRNA primer captured by HIV-1 and MuLV occurs after nuclear export of tRNA and supports a model in which primer selection for retroviruses is coordinated with tRNA biogenesis at the intracellular site of protein synthesis. PMID- 12885889 TI - Oral immunogenicity of human papillomavirus-like particles expressed in potato. AB - Human papillomavirus-like particles (HPV VLPs) have shown considerable promise as a parenteral vaccine for the prevention of cervical cancer and its precursor lesions. Parenteral vaccines are expensive to produce and deliver, however, and therefore are not optimal for use in resource-poor settings, where most cervical HPV disease occurs. Transgenic plants expressing recombinant vaccine immunogens offer an attractive and potentially inexpensive alternative to vaccination by injection. For example, edible plants can be grown locally and can be distributed easily without special training or equipment. To assess the feasibility of an HPV VLP-based edible vaccine, in this study we synthesized a plant codon-optimized version of the HPV type 11 (HPV11) L1 major capsid protein coding sequence and introduced it into tobacco and potato. We show that full-length L1 protein is expressed and localized in plant cell nuclei and that expression of L1 in plants is enhanced by removal of the carboxy-terminal nuclear localization signal sequence. We also show that plant-expressed L1 self-assembles into VLPs with immunological properties comparable to those of native HPV virions. Importantly, ingestion of transgenic L1 potato was associated with activation of an anti-VLP immune response in mice that was qualitatively similar to that induced by VLP parenteral administration, and this response was enhanced significantly by subsequent oral boosting with purified insect cell-derived VLPs. Thus, papillomavirus L1 protein can be expressed in transgenic plants to form immunologically functional VLPs, and ingestion of such material can activate potentially protective humoral immune responses. PMID- 12885890 TI - Replication-competent bacterial artificial chromosomes of Marek's disease virus: novel tools for generation of molecularly defined herpesvirus vaccines. AB - Marek's disease (MD), a highly infectious disease caused by an oncogenic herpesvirus, is one of the few herpesvirus diseases against which live attenuated vaccines are used as the main strategy for control. We have constructed bacterial artificial chromosomes (BACs) of the CVI988 (Rispens) strain of the virus, the most widely used and effective vaccine against MD. Viruses derived from the BAC clones were stable after in vitro and in vivo passages and showed characteristics and growth kinetics similar to those of the parental virus. Molecular analysis of the individual BAC clones showed differences in the structure of the meq gene, indicating that the commercial vaccine contains virus populations with distinct genomic structures. We also demonstrate that, contrary to the published data, the sequence of the L-meq of the BAC clone did not show any frameshift. Virus stocks derived from one of the BAC clones (clone 10) induced 100 percent protection against infection by the virulent strain RB1B, indicating that BAC-derived viruses could be used with efficacies similar to those of the parental CVI988 vaccines. As a DNA vaccine, this BAC clone was also able to induce protection in 6 of 20 birds. Isolation of CVI988 virus from all of these six birds suggested that immunity against challenge was probably dependent on the reconstitution of the virus in vivo and that such viruses are also as immunogenic as the in vitro grown BAC-derived or parental vaccine viruses. Although the reasons for the induction of protection only in a proportion of birds (33.3%) that received the DNA vaccine are not clear, this is most likely to be related to the suboptimal method of DNA delivery. The construction of the CVI988 BAC is a major step towards understanding the superior immunogenic features of CVI988 and provides the opportunity to exploit the power of BAC technology for generation of novel molecularly defined vaccines. PMID- 12885891 TI - Induction of protective immune responses against R5 human immunodeficiency virus type 1 (HIV-1) infection in hu-PBL-SCID mice by intrasplenic immunization with HIV-1-pulsed dendritic cells: possible involvement of a novel factor of human CD4(+) T-cell origin. AB - The potential of a dendritic cell (DC)-based vaccine against human immunodeficiency virus type 1 (HIV-1) infection in humans was explored with SCID mice reconstituted with human peripheral blood mononuclear cells (PBMC). HIV-1 negative normal human PBMC were transplanted directly into the spleens of SCID mice (hu-PBL-SCID-spl mice) together with autologous mature DCs pulsed with either inactivated HIV-1 (strain R5 or X4) or ovalbumin (OVA), followed by a booster injection 5 days later with autologous DCs pulsed with the same respective antigens. Five days later, these mice were challenged intraperitoneally with R5 HIV-1(JR-CSF). Analysis of infection at 7 days postinfection showed that the DC-HIV-1-immunized hu-PBL-SCID-spl mice, irrespective of the HIV-1 isolate used for immunization, were protected against HIV-1 infection. In contrast, none of the DC-OVA-immunized mice were protected. Sera from the DC-HIV-1- but not the DC-OVA-immunized mice inhibited the in vitro infection of activated PBMC and macrophages with R5, but not X4, HIV-1. Upon restimulation with HIV-1 in vitro, the human CD4(+) T cells derived from the DC HIV-1-immunized mice produced a similar R5 HIV-1 suppressor factor. Neutralizing antibodies against human RANTES, MIP-1alpha, MIP-1beta, alpha interferon (IFN alpha), IFN-beta, IFN-gamma, interleukin-4 (IL-4), IL-10, IL-13, IL-16, MCP-1, MCP-3, tumor necrosis factor alpha (TNF-alpha), or TNF-beta failed to reverse the HIV-1-suppressive activity. These results show that inactivated HIV-1-pulsed autologous DCs can stimulate splenic resident human CD4(+) T cells in hu-PBL-SCID spl mice to produce a yet-to-be-defined, novel soluble factor(s) with protective properties against R5 HIV-1 infection. PMID- 12885892 TI - Plasmid chemokines and colony-stimulating factors enhance the immunogenicity of DNA priming-viral vector boosting human immunodeficiency virus type 1 vaccines. AB - Heterologous "prime-boost" regimens that involve priming with plasmid DNA vaccines and boosting with recombinant viral vectors have been shown to elicit potent virus-specific cytotoxic T-lymphocyte responses. Increasing evidence, however, suggests that the utility of recombinant viral vectors in human populations will be significantly limited by preexisting antivector immunity. Here we demonstrate that the coadministration of plasmid chemokines and colony stimulating factors with plasmid DNA vaccines markedly increases the immunogenicity of DNA prime-recombinant adenovirus serotype 5 (rAd5) boost and DNA prime-recombinant vaccinia virus (rVac) boost vaccine regimens in BALB/c mice. In mice with preexisting anti-Ad5 immunity, priming with the DNA vaccine alone followed by rAd5 boosting elicited only marginal immune responses. In contrast, cytokine-augmented DNA vaccine priming followed by rAd5 vector boosting was able to generate potent immune responses in mice with preexisting anti-Ad5 immunity. These data demonstrate that plasmid cytokines can markedly improve the immunogenicity of DNA prime-viral vector boost vaccine strategies and can partially compensate for antivector immunity. PMID- 12885893 TI - Identification of the E9/E2C cDNA and functional characterization of the gene product reveal a new repressor of transcription and replication in cottontail rabbit papillomavirus. AB - Cottontail rabbit papillomavirus (CRPV) genomes mutated in the trans-activation domain of the E2 protein, which stimulates both viral DNA replication and transcription, are severely impaired in their ability to induce tumors in New Zealand White rabbits. A number of papillomaviruses encode, in addition to full length E2, a shortened E2 protein or an E2 protein fused to a short stretch of amino acids derived from the small E8 open reading frame that counteract the activities of E2. We identified and cloned the novel cDNA E9/E2C of CRPV from papillomas of New Zealand White and cottontail rabbits and characterized the functions of the encoded gene product. E9/E2C was shown to be a bona fide repressor of minimal viral promoters, with the E9 domain being essential for this activity, and to repress E1/E2-dependent replication of a CRPV origin construct. In addition, E9/E2C counteracted the transactivation effect of the full-length E2 on minimal promoters containing several E2 binding sites. To investigate the role of E9/E2C in tumorigenesis, we constructed two CRPV genomes mutated in E9/E2C. One, designated CRPV-E9atgmut-pLAII, contained a mutation in the unique start codon in the E9 open reading frame, and the second E9/E2C mutant was constructed by the introduction of a stop codon close to the splice donor site at nucleotide 3714 that additionally prevented the correct splicing of the transcript. When we infected New Zealand White rabbits with these constructs, we surprisingly noted no differences in tumor induction efficiency, viral genome copy number, and viral transcription in comparison to wild-type CRPV. PMID- 12885894 TI - Stable high-producer cell clone expressing virus-like particles of the Japanese encephalitis virus e protein for a second-generation subunit vaccine. AB - We produced and characterized a cell clone (J12#26 cells) that stably expresses Japanese encephalitis virus (JEV) cDNA, J12, which encodes the viral signal peptide, premembrane (prM), and envelope (E) proteins (amino acid positions 105 to 794). Rabbit kidney-derived RK13 cells were transfected with a J12 expression plasmid, selected by resistance to marker antibiotics, and cloned by two cycles of a limiting-dilution method in the presence of antibiotics, a procedure that prevents the successful generation of E-producing cell clones. J12#26 cells secreted virus-like particles containing the authentic E antigen (E-VLP) into the culture medium in a huge enzyme-linked immunosorbent assay-equivalent amount (2.5 micro g per 10(4) cells) to the internationally licensed JE vaccine JE-VAX. E-VLP production was stable after multiple cell passages and persisted over 1 year with 100% expressing cells without detectable cell fusion, apoptosis, or cell death, but was suspended when the cells grew to 100% confluency and contact inhibition occurred. Mice immunized with the purified J12#26 E-antigen without adjuvant developed high titers of neutralizing antibodies for at least 7 months and 100% protection against intraperitoneal challenge with 5 x 10(6) PFU of JEV when examined according to the JE vaccine standardization protocol. These results suggest that the recombinant E-VLP antigen produced by the J12#26 cell clone is an effective, safe, and low-cost second-generation subunit JE vaccine. PMID- 12885895 TI - Probing the importance of tRNA anticodon: human immunodeficiency virus type 1 (HIV-1) RNA genome complementarity with an HIV-1 that selects tRNA(Glu) for replication. AB - The initiation of human immunodeficiency virus type 1 (HIV-1) reverse transcription occurs at the primer binding site (PBS) that is complementary to the 3'-terminal nucleotides of tRNA(3)(Lys). Why all known strains of HIV-1 select tRNA(3)(Lys) for replication is unknown. Previous studies on the effect of altering the PBS of HIV-1 on replication identified an HIV-1 with a PBS complementary to tRNA(Glu). Since the virus was not initially designed to use tRNA(Glu), the virus had selected tRNA(Glu) from the intracellular pool of tRNA for use in replication. Further characterization of HIV-1 that uses tRNA(Glu) may provide new insights into the preference for tRNA(3)(Lys). HIV-1 constructed with the PBS complementary to tRNA(Glu) was more stable than HIV-1 with the PBS complementary to tRNA(Met) or tRNA(His); however, all of these viruses eventually reverted back to using tRNA(3)(Lys) following growth in SupT1 cells or peripheral blood mononuclear cells (PBMCs). New HIV-1 mutants with nucleotides in U5 complementary to the anticodon of tRNA(Glu) remained stable when grown in SupT1 cells or PBMCs, although the mutants grew more slowly than the wild-type virus. Sequence analysis of the U5 region and the PBS revealed additional mutations predicted to further promote tRNA-viral genome interaction. The results support the importance of the tRNA anticodon-genome interaction in the selection of the tRNA primer and highlight the fact that unique features of tRNA(3)(Lys) are exploited by HIV-1 for selection as the reverse transcription primer. PMID- 12885896 TI - A hepadnavirus regulatory element enhances expression of a type 2 bovine viral diarrhea virus E2 protein from a bovine herpesvirus 1 vector. AB - Recently, the possibility of using virus vectors to immunize cattle against selected bovine viral diarrhea virus (BVDV) genes has gained widespread interest. However, when we attempted to express the E2 protein from type 2 (890 strain) BVDV in a bovine herpesvirus 1 (BHV1) vector, we observed that expression was poor. This often happens when genes from a cytoplasmic virus are expressed in the cell nucleus. To counter this effect, we attempted to enhance expression by a strategy employed by viruses. RNAs of retroviruses and hepadnaviruses contain cis acting elements that facilitate expression of RNAs that otherwise are degraded or retained within the nucleus. In Mason-Pfizer monkey virus, the required RNA sequence element is known as a constitutive transport element (CTE). A related element from woodchuck hepatitis virus is known as the woodchuck posttranscriptional regulatory element (WPRE). We tested the ability of the CTE, the WPRE, and introns to enhance expression of E2. All three elements stimulated expression of E2 from plasmids. The combination of the WPRE and an intron yielded the highest level of E2 expression in plasmids. However, when E2 was expressed from a BHV1 vector, the presence of an intron was inhibitory. In contrast, the WPRE was very efficient at stimulating E2 expression from a BHV1 vector. This result represents the first expression of a type 2 BVDV E2 protein from a mammalian virus vector and raises the possibility that the WPRE may provide a general method of enhancing foreign gene expression from BHV1 and other herpesvirus vectors. PMID- 12885897 TI - Simian immunodeficiency virus infection in neonatal macaques. AB - Children with human immunodeficiency virus infection often have higher viral loads and progress to AIDS more rapidly than adults. Since the intestinal tract is a major site of early viral replication and CD4(+) T-cell depletion in adults, we examined the effects of simian immunodeficiency virus (SIV) on both peripheral and intestinal lymphocytes from 13 neonatal macaques infected with SIVmac239. Normal neonates had more CD4(+) T cells and fewer CD8(+) T cells in all tissues than adults. Surprisingly, neonates had substantial percentages of CD4(+) T cells with an activated, memory phenotype (effector CD4(+) T cells) in the lamina propria of the intestine compared to peripheral lymphoid tissues, even when examined on the day of birth. Moreover, profound and selective depletion of jejunum lamina propria CD4(+) T cells occurred in neonatal macaques within 21 days of infection, which was preceded by large numbers of SIV-infected cells in this compartment. Furthermore, neonates with less CD4(+) T-cell depletion in tissues tended to have higher viral loads. The persistence of intestinal lamina propria CD4(+) T cells in some neonates with high viral loads suggests that increased turnover and/or resistance to CD4(+) T-cell loss may contribute to the higher viral loads and increased severity of disease in neonatal hosts. PMID- 12885898 TI - Adaptation of Puumala hantavirus to cell culture is associated with point mutations in the coding region of the L segment and in the noncoding regions of the S segment. AB - We previously developed a model for studies on hantavirus host adaptation and initiated genetic analysis of Puumala virus variants passaged in colonized bank voles and in cultured Vero E6 cells. With the data presented in this paper, the sequence comparison of the wild-type and Vero E6-adapted variants of Puumala virus, strain Kazan, has been completed. The only amino acid substitution that distinguished the two virus variants was found in the L protein, Ser versus Phe at position 2053. Another mutation found in the L segment, the silent transition C1053U, could result from the selection of a variant with altered L RNA folding. Nucleotide substitutions observed in individual L cDNA clones, most of them A-->G and U-->C transitions, suggested that the population of L RNA molecules is represented by quasispecies. The mutation frequency in the L segment quasispecies appeared to be similar to the corresponding values for the S and M quasispecies. Analysis of the cDNA clones with the complete S segment sequences from passage 20 confirmed our earlier conclusion that the cell-adapted genotype of the virus is represented mostly by variants with mutated S segment noncoding regions. However, the spectrum of the S segment quasispecies appeared to be changing, suggesting that, after the initial adaptation (passages 1 to 11), the viral population is still being driven by selection for variants with higher fitness. PMID- 12885899 TI - The coronavirus spike protein is a class I virus fusion protein: structural and functional characterization of the fusion core complex. AB - Coronavirus entry is mediated by the viral spike (S) glycoprotein. The 180-kDa oligomeric S protein of the murine coronavirus mouse hepatitis virus strain A59 is posttranslationally cleaved into an S1 receptor binding unit and an S2 membrane fusion unit. The latter is thought to contain an internal fusion peptide and has two 4,3 hydrophobic (heptad) repeat regions designated HR1 and HR2. HR2 is located close to the membrane anchor, and HR1 is some 170 amino acids (aa) upstream of it. Heptad repeat (HR) regions are found in fusion proteins of many different viruses and form an important characteristic of class I viral fusion proteins. We investigated the role of these regions in coronavirus membrane fusion. Peptides HR1 (96 aa) and HR2 (39 aa), corresponding to the HR1 and HR2 regions, were produced in Escherichia coli. When mixed together, the two peptides were found to assemble into an extremely stable oligomeric complex. Both on their own and within the complex, the peptides were highly alpha helical. Electron microscopic analysis of the complex revealed a rod-like structure approximately 14.5 nm in length. Limited proteolysis in combination with mass spectrometry indicated that HR1 and HR2 occur in the complex in an antiparallel fashion. In the native protein, such a conformation would bring the proposed fusion peptide, located in the N-terminal domain of HR1, and the transmembrane anchor into close proximity. Using biological assays, the HR2 peptide was shown to be a potent inhibitor of virus entry into the cell, as well as of cell-cell fusion. Both biochemical and functional data show that the coronavirus spike protein is a class I viral fusion protein. PMID- 12885900 TI - Strain variation in an emerging iridovirus of warm-water fishes. AB - Although iridoviruses vary widely within and among genera with respect to their host range and virulence, variation within iridovirus species has been less extensively characterized. This study explores the nature and extent of intraspecific variation within an emerging iridovirus of North American warm water fishes, largemouth bass virus (LMBV). Three LMBV isolates recovered from three distinct sources differed genetically and phenotypically. Genetically, the isolates differed in the banding patterns generated from amplified fragment length polymorphism analysis but not in their DNA sequences at two loci of different degrees of evolutionary stability. In vitro, the isolates replicated at identical rates in cell culture, as determined by real-time quantitative PCR of viral particles released into suspension. In vivo, the isolates varied over fivefold in virulence, as measured by the rate at which they induced mortality in juvenile largemouth bass. This variation was reflected in the viral loads of exposed fish, measured using real-time quantitative PCR; the most virulent viral strain also replicated to the highest level in fish. Together, these results justify the designation of these isolates as different strains of LMBV. Strain variation in iridoviruses could help explain why animal populations naturally infected with iridovirus pathogens vary so extensively in their clinical responses to infection. The results of this study are especially relevant to emerging iridoviruses of aquaculture systems and wildlife. PMID- 12885901 TI - Feline immunodeficiency virus ORF-Ais required for virus particle formation and virus infectivity. AB - The orf-A (orf-2) gene of feline immunodeficiency virus (FIV) is a small open reading frame predicted to encode a 77-amino-acid protein that contains putative domains similar to those of the ungulate lentiviral Tat protein. Orf-A is reported to be critical for efficient viral replication in vitro and in vivo. A series of FIV-pPPR-derived proviruses with in-frame deletions and point mutations within orf-A were constructed and tested for replication in feline lymphoid cells. Orf-A mutant proviruses were also tested for viral gene and protein expression, viral particle formation, and virion infectivity. Deletions within orf-A severely restricted FIV replication in feline peripheral blood mononuclear cells (PBMC) and interleukin-2-dependent T-cell lines. In addition, substitutions of alanines for leucines in the putative leucine-rich domain, for cysteines in the putative cysteine-rich domain, and for a tryptophan at position 43 in Orf-A restricted the replication of FIV mutants. Deletions and point mutations in orf-A imposed a small effect or no effect on FIV long-terminal-repeat-driven viral gene expression and had no effect on viral protein expression. However, release of cell-free, virion-associated viral RNA in supernatants from cells transfected with orf-A mutant proviruses was severely restricted but was rescued by cotransfection with a wild-type Orf-A expression vector. In addition, virions derived from orf-A mutant proviruses expressed reduced infectivity for feline PBMC. Our findings suggest that Orf-A functions involve multiple steps of the FIV life cycle including both virion formation and infectivity. Furthermore, these observations suggest that Orf-A represents an FIV-encoded analog more similar to the accessory gene vpr, vpu, or nef than to the regulatory gene tat encoded by the primate lentiviruses. PMID- 12885902 TI - De novo synthesis of negative-strand RNA by Dengue virus RNA-dependent RNA polymerase in vitro: nucleotide, primer, and template parameters. AB - By using a purified dengue virus RNA-dependent RNA polymerase and a subgenomic 770-nucleotide RNA template, it was shown previously that the ratio of the de novo synthesis product to hairpin product formed was inversely proportional to increments of assay temperatures (20 to 40 degrees C). In this study, the components of the de novo preinitiation complex are defined as ATP, a high concentration of GTP (500 micro M), the polymerase, and the template RNA. Even when the 3'-terminal sequence of template RNA was mutated from -GGUUCU-3' to GGUUUU-3', a high GTP concentration was required for de novo initiation, suggesting that high GTP concentration plays a conformational role. Furthermore, utilization of synthetic primers by the polymerase indicated that AGAA is the optimal primer whereas AG, AGA, and AGAACC were inefficient primers. Moreover, mutational analysis of the highly conserved 3'-terminal dinucleotide CU of the template RNA indicated that change of the 3'-terminal nucleotide from U to C reduced the efficiency about fivefold. The order of preference for the 3' terminal nucleotide, from highest to lowest, is U, A - G, and C. However, change of the penultimate nucleotide from C to U did not affect the template activity. A model consistent with these results is that the active site of the polymerase switches from a "closed" form, catalyzing de novo initiation through synthesis of short primers, to an "open" form for elongation of a double-stranded template primer. PMID- 12885903 TI - Development of recombinant vesicular stomatitis viruses that exploit defects in host defense to augment specific oncolytic activity. AB - Vesicular stomatitis virus (VSV) is a negative-stranded RNA virus normally sensitive to the antiviral actions of alpha/beta interferon (IFN-alpha/beta). Recently, we reported that VSV replicates to high levels in many transformed cells due, in part, to susceptible cells harboring defects in the IFN system. These observations were exploited to demonstrate that VSV can be used as a viral oncolytic agent to eradicate malignant cells in vivo while leaving normal tissue relatively unaffected. To attempt to improve the specificity and efficacy of this system as a potential tool in gene therapy and against malignant disease, we have genetically engineered VSV that expresses the murine IFN-beta gene. The resultant virus (VSV-IFNbeta) was successfully propagated in cells not receptive to murine IFN-alpha/beta and expressed high levels of functional heterologous IFN-beta. In normal murine embryonic fibroblasts (MEFs), the growth of VSV-IFNbeta was greatly reduced and diminished cytopathic effect was observed due to the production of recombinant IFN-beta, which by functioning in a manner involving autocrine and paracrine mechanisms induced an antiviral effect, preventing virus growth. However, VSV-IFNbeta grew to high levels and induced the rapid apoptosis of transformed cells due to defective IFN pathways being prevalent and thus unable to initiate proficient IFN-mediated host defense. Importantly, VSV expressing the human IFN-beta gene (VSV-hIFNbeta) behaved comparably and, while nonlytic to normal human cells, readily killed their malignant counterparts. Similar to our in vitro observations, following intravenous and intranasal inoculation in mice, recombinant VSV (rVSV)-IFNbeta was also significantly attenuated compared to wild type VSV or rVSV expressing green fluorescent protein. However, VSV-IFNbeta retained propitious oncolytic activity against metastatic lung disease in immunocompetent animals and was able to generate robust antitumor T-cell responses. Our data indicate that rVSV designed to exploit defects in mechanisms of host defense can provide the basis for new generations of effective, specific, and safer viral vectors for the treatment of malignant and other disease. PMID- 12885904 TI - Investigation of structural and functional motifs within the vaccinia virus A14 phosphoprotein, an essential component of the virion membrane. AB - We have previously reported the construction and characterization of an inducible recombinant virus in which expression of the vaccinia virus membrane protein A14 is experimentally regulated using the tetracycline operator-repressor system. Repression of A14, which results in a 1,000-fold reduction in viral yield, leads to an early block in viral morphogenesis characterized by the accumulation of large virosomes, empty "crescents" that fail to contact these virosomes, and, most strikingly, large numbers of aberrant 25-nm vesicles. Here we report the establishment of a transient-complementation system for the structure-function analysis of A14. We have constructed numerous mutant alleles of A14 designed to identify and test the importance of key structural and sequence motifs within A14, including sites of posttranslational modification, such as glycosylation, phosphorylation, and dimerization. From these studies we have determined that robust complementation ability requires an intact N terminus and two regions flanking the first membrane-spanning domain of A14. We show that A14 is modified by N-linked glycosylation both in vitro and in vivo. However, only a minority of A14 molecules are glycosylated in vivo and these are not encapsidated. In this report we also identify the sole phosphorylated serine residue of A14 as lying within the NHS(85) motif that undergoes glycosylation. Additionally, we show that the Cys(71) residue is required for intermolecular disulfide bond formation and describe the properties of a virus expressing an allele of A14 that cannot form disulfide-linked dimers. PMID- 12885905 TI - Deletions in the putative cell receptor-binding domain of Sindbis virus strain MRE16 E2 glycoprotein reduce midgut infectivity in Aedes aegypti. AB - The Sindbis virus (Alphavirus; Togaviridae) strain MRE16 efficiently infects Aedes aegypti mosquitoes that ingest a blood meal containing 8 to 9 log(10) PFU of virus/ml. However, a small-plaque variant of this virus, MRE16sp, poorly infects mosquitoes after oral infection with an equivalent titer. To determine the genetic differences between MRE16 and MRE16sp viruses, we have sequenced the MRE16sp structural genes and found a 90-nucleotide deletion in the E2 glycoprotein that spans the 3' end of the coding region for the putative cell receptor binding domain (CRBD). We examined the role of this deletion in oral infection of mosquitoes by constructing infectious clones pMRE16icDeltaE200-Y229 and pMRE16ic, representing MRE16 virus genomes with and without the deletion, respectively. A third infectious clone, pMRE16icDeltaE200-C220, was also constructed that contained a smaller deletion extending only to the 3' terminus of the CRBD coding region. Virus derived from pMRE16ic replicated with the same efficiency as parental virus in vertebrate (BHK-21) and mosquito (C6/36) cells and orally infected A. aegypti. Viruses derived from pMRE16icDeltaE200-Y229 and pMRE16icDeltaE200-C220 replicated 10- to 100-fold less efficiently in C6/36 and BHK-21 cells than did MRE16ic virus. Each deletion mutant poorly infected A. aegypti and dramatically reduced midgut infectivity and dissemination. However, all viruses generated nearly equal titers (approximately 6.0 log(10) PFU/ml) in mosquitoes 4 days after infection by intrathoracic inoculation. These results suggest that the deleted portion of the E2 CRBD represents an important determinant of MRE16 virus midgut infectivity in A. aegypti. PMID- 12885906 TI - Endocytosis of hepatitis B immune globulin into hepatocytes inhibits the secretion of hepatitis B virus surface antigen and virions. AB - Hepatitis B immunoglobulin is used for prophylaxis against hepatitis B virus (HBV) and is thought to act by neutralization of virions and hepatitis B virus surface antigen (HBsAg)-containing particles in circulation. Using a panel of hepatocyte-derived cell lines, the present study investigated in vitro whether HBs-specific immunoglobulin G (IgG) is internalized in hepatocytes and whether it interacts with HBsAg in the cells. By immunoelectron microscopy and immunoblotting, human IgG and FcRn receptor for IgG were demonstrated on cellular membranes and in cytoplasmic extracts, irrespective of the HBsAg status of the cells. Furthermore, HBsAg and anti-HBs were shown to be colocalized in the same cellular compartment by two-color confocal microscopy. Endocytosis of HBs specific IgG caused intracellular accumulation of HBsAg in a dose-dependent manner and inhibited the secretion of HBsAg and HBV virions from the cells. These effects were not observed with F(ab)(2) fragments or nonimmune IgG as controls. The specificity of intracellular HBsAg- anti-HBs interaction was further investigated in cells transfected with HBV genomes expressing wild-type HBsAg or immune escape HBsAg (with a G145R mutation). Monoclonal anti-HBs markedly reduced the secretion of wild-type HBsAg, while the secretion of mutant HBsAg was not affected. These results suggest that HBs-specific IgG binds to hepatocytes and interacts with HBsAg within the cells. This may be relevant for the selection of surface antibody escape mutations. PMID- 12885907 TI - Cell cycle arrest by Kaposi's sarcoma-associated herpesvirus replication associated protein is mediated at both the transcriptional and posttranslational levels by binding to CCAAT/enhancer-binding protein alpha and p21(CIP-1). AB - Lytic-cycle replication of Kaposi's sarcoma-associated herpesvirus (KSHV) in PEL cells causes G(1) cell cycle arrest mediated by the virus-encoded replication associated protein (RAP) (or K8 protein), which induces high-level expression of the cellular C/EBPalpha and p21 proteins. Here we have examined the mechanism of this induction at both the transcriptional and posttranslational levels. RAP proved to bind very efficiently to both C/EBPalpha and p21 and stabilized them by up to 10-fold from proteasome-mediated degradation in vitro. Cross-linking revealed that RAP itself forms stable dimers and tetramers in solution and forms higher-order complexes but not heterodimers with C/EBPalpha. Cotransfection of RAP with C/EBPalpha cooperatively stimulated both the C/EBPalpha and p21 promoters in luciferase reporter gene assays. Only the basic/leucine zipper region of RAP was needed for interaction with and stabilization of C/EBPalpha, but both the N-terminal and C-terminal domains were required for transcriptional augmentation. In vitro-translated RAP interfered with DNA binding by C/EBPalpha in electrophonetic mobility shift assay (EMSA) experiments but did not itself bind to the target C/EBPalpha sites or form supershifted bands. However, in endogenous chromatin immunoprecipitation (ChIP) assays with tetradecanoyl phorbol acetate-induced PEL cells, RAP proved to specifically associate with the C/EBPalpha promoter in vivo, but only in a C/EBPalpha-dependent manner, implying an in vivo piggyback interaction with DNA-bound C/EBPalpha. Expression of exogenous RAP (Ad-RAP) caused G(1)/S cell cycle arrest in human dermal microvascular endothelial cells and also induced both the C/EBPalpha and p21 proteins, which formed punctate nuclear patterns that colocalized with RAP in PML nuclear bodies. In the presence of RAP, C/EBPalpha was also efficiently recruited into viral DNA replication compartments in both infected and cotransfected cells. In support of a direct role for this interaction in viral DNA replication, three C/EBPalpha binding sites were identified by in vitro EMSA experiments within a 220-bp core segment of the duplicated KSHV Ori-Lyt region, and although RAP did not bind to Ori-Lyt DNA directly in vitro, both endogenous RAP and C/EBPalpha were found to be associated with the Ori-Lyt region by ChIP assays in lytically induced PEL cells. Finally, we found that the KSHV lytic cycle could not be triggered by either synchronizing KSHV latently infected PEL cells in G(1) phase or inducing p21 in a C/EBPalpha-independent process. PMID- 12885908 TI - Residues in the apical domain of the feline and canine transferrin receptors control host-specific binding and cell infection of canine and feline parvoviruses. AB - Canine parvovirus (CPV) and feline panleukopenia virus (FPV) capsids bind to the transferrin receptors (TfRs) of their hosts and use these receptors to infect cells. The binding is partially host specific, as FPV binds only to the feline TfR, while CPV binds to both the canine and feline TfRs. The host-specific binding is controlled by a combination of residues within a raised region of the capsid. To define the TfR structures that interact with the virus, we altered the apical domain of the feline or canine TfR or prepared chimeras of these receptors and tested the altered receptors for binding to FPV or CPV capsids. Most changes in the apical domain of the feline TfR did not affect binding, but replacing Leu221 with Ser or Asp prevented receptor binding to either FPV or CPV capsids, while replacing Leu221 with Lys resulted in a receptor that bound only to CPV but not to FPV. Analysis of recombinants of the feline and canine TfRs showed that sequences controlling CPV-specific binding were within the apical domain and that more than one difference between these receptors determined the CPV-specific binding of the canine TfR. Single changes within the canine TfR which removed a single amino acid insertion or which eliminated a glycosylation site gave that receptor the expanded ability to bind to FPV and CPV. In some cases, binding of capsids to mutant receptors did not result in infection, suggesting a structural role for the receptor in cell infection by the viruses. PMID- 12885909 TI - Incorporation of tick-borne encephalitis virus replicons into virus-like particles by a packaging cell line. AB - RNA replicons derived from flavivirus genomes show considerable potential as gene transfer and immunization vectors. A convenient and efficient encapsidation system is an important prerequisite for the practical application of such vectors. In this work, tick-borne encephalitis (TBE) virus replicons and an appropriate packaging cell line were constructed and characterized. A stable CHO cell line constitutively expressing the two surface proteins prM/M and E (named CHO-ME cells) was generated and shown to efficiently export mature recombinant subviral particles (RSPs). When replicon NdDeltaME lacking the prM/M and E genes was introduced into CHO-ME cells, virus-like particles (VLPs) capable of initiating a single round of infection were released, yielding titers of up to 5 x 10(7)/ml in the supernatant of these cells. Another replicon (NdDeltaCME) lacking the region encoding most of the capsid protein C in addition to proteins prM/M and E was not packaged by CHO-ME cells. As observed with other flavivirus replicons, both TBE virus replicons appeared to exert no cytopathic effect on their host cells. Sedimentation analysis revealed that the NdDeltaME-containing VLPs were physically distinct from RSPs and similar to infectious virions. VLPs could be repeatedly passaged in CHO-ME cells but maintained the property of being able to initiate only a single round of infection in other cells during these passages. CHO-ME cells can thus be used both as a source for mature TBE virus RSPs and as a safe and convenient replicon packaging cell line, providing the TBE virus surface proteins prM/M and E in trans. PMID- 12885910 TI - Reovirus-induced alteration in expression of apoptosis and DNA repair genes with potential roles in viral pathogenesis. AB - Reoviruses are a leading model for understanding cellular mechanisms of virus induced apoptosis. Reoviruses induce apoptosis in multiple cell lines in vitro, and apoptosis plays a key role in virus-induced tissue injury of the heart and brain in vivo. The activation of transcription factors NF-kappaB and c-Jun are key events in reovirus-induced apoptosis, indicating that new gene expression is critical to this process. We used high-density oligonucleotide microarrays to analyze cellular transcriptional alterations in HEK293 cells after infection with reovirus strain T3A (i.e., apoptosis inducing) compared to infection with reovirus strain T1L (i.e., minimally apoptosis inducing) and uninfected cells. These strains also differ dramatically in their potential to induce apoptotic injury in hearts of infected mice in vivo-T3A is myocarditic, whereas T1L is not. Using high-throughput microarray analysis of over 12,000 genes, we identified differential expression of a defined subset of genes involved in apoptosis and DNA repair after reovirus infection. This provides the first comparative analysis of altered gene expression after infection with viruses of differing apoptotic phenotypes and provides insight into pathogenic mechanisms of virus-induced disease. PMID- 12885911 TI - Local delivery of CpG oligodeoxynucleotides induces rapid changes in the genital mucosa and inhibits replication, but not entry, of herpes simplex virus type 2. AB - Mucosal surfaces are the entry sites for the vast majority of infectious pathogens and provide the first line of defense against infection. In addition to the epithelial barrier, the innate immune system plays a key role in recognizing and rapidly responding to invading pathogens via innate receptors, such as Toll like receptors (TLR). Bacterial CpG DNA, a potent activator of innate immunity, is recognized by TLR9. Here, we confirm that local mucosal, but not systemic, delivery of CpG oligodeoxynucleotides (ODN) to the genital tract protects mice from a subsequent lethal vaginal herpes simplex virus type 2 (HSV-2) challenge. Since these effects were so local in action, we examined the genital mucosa. Local delivery of CpG ODN induced rapid proliferation and thickening of the genital epithelium and caused significant recruitment of inflammatory cells to the submucosa. Local CpG ODN treatment also resulted in inhibition of HSV-2 replication but had no effect on HSV-2 entry into the genital mucosa. CpG ODN induced protection against HSV-2 was not associated with early increases in gamma interferon (IFN-gamma) secretion in the genital tract, and CpG ODN-treated IFN gamma(-/-) mice were protected from subsequent challenge with a lethal dose of HSV-2. Treatment of human HEK-293 cells transfected with murine TLR9 showed that the antiviral activity of CpG ODN was mediated through TLR9. These studies suggest that local induction of mucosal innate immunity can provide protection against sexually transmitted infections, such as HSV-2 or possibly human immunodeficiency virus, at the mucosal surfaces. PMID- 12885912 TI - Conditional suppression of cellular genes: lentivirus vector-mediated drug inducible RNA interference. AB - RNA interference has emerged as a powerful technique to downregulate the expression of specific genes in cells and in animals, thus opening new perspectives in fields ranging from developmental genetics to molecular therapeutics. Here, we describe a method that significantly expands the potential of RNA interference by permitting the conditional suppression of genes in mammalian cells. Within a lentivirus vector background, we subjected the polymerase III promoter-dependent production of small interfering RNAs to doxycycline-controllable transcriptional repression. The resulting system can achieve the highly efficient and completely drug-inducible knockdown of cellular genes. As lentivirus vectors can stably transduce a wide variety of targets both in vitro and in vivo and can be used to generate transgenic animals, the present system should have broad applications. PMID- 12885914 TI - Complete genomic sequencing shows that polioviruses and members of human enterovirus species C are closely related in the noncapsid coding region. AB - The 65 human enterovirus serotypes are currently classified into five species: Poliovirus (3 serotypes), Human enterovirus A (HEV-A) (12 serotypes), HEV-B (37 serotypes), HEV-C (11 serotypes), and HEV-D (2 serotypes). Coxsackie A virus (CAV) serotypes 1, 11, 13, 15, 17, 18, 19, 20, 21, 22, and 24 constitute HEV-C. We have determined the complete genome sequences for the remaining nine HEV-C serotypes and compared them with the complete sequences of CAV21, CAV24, and the polioviruses. The viruses were most diverse in the capsid region (4 to 36% amino acid difference). A high degree of capsid sequence conservation (96% amino acid identity) suggests that CAV15 and CAV18 should be classified as strains of CAV11 and CAV13, respectively. In the 3CD region, CAV1, CAV19, and CAV22 differed from one another by only 1.2 to 1.4% and CAV11, CAV13, CAV17, CAV20, CAV21, CAV24, and the polioviruses differed from one another by only 1.2 to 3.6%. The two groups, however, differed from one another by 14.6 to 16.2%. The polioviruses as a group were monophyletic only in the capsid region. Only one group of serotypes (CAV1, CAV19, and CAV22) was consistently monophyletic in multiple genome regions. Incongruities among phylogenetic trees based on different genome regions strongly suggest that recombination has occurred between the polioviruses, CAV11, CAV13, CAV17, and CAV20. The close relationship among the polioviruses and CAV11, CAV13, CAV17, CAV20, CAV21, and CAV24 and the uniqueness of CAV1, CAV19, and CAV22 suggest that revisions should be made to the classification of these viruses. PMID- 12885913 TI - Function of herpes simplex virus type 1 gD mutants with different receptor binding affinities in virus entry and fusion. AB - We have studied the receptor-specific function of four linker-insertion mutants of herpes simplex virus type 1 glycoprotein D (gD) representing each of the functional regions of gD. We used biosensor analysis to measure binding of the gD mutants to the receptors HVEM (HveA) and nectin-1 (HveC). One of the mutants, gD(inverted Delta 34t), failed to bind HVEMt but showed essentially wild-type (WT) affinity for nectin-1t. The receptor-binding kinetics and affinities of the other three gD mutants varied over a 1,000-fold range, but each mutant had the same affinity for both receptors. All of the mutants were functionally impaired in virus entry and cell fusion, and the levels of activity were strikingly similar in these two assays. gD(inverted Delta 34)-containing virus was defective on HVEM-expressing cells but did enter nectin-1-expressing cells to about 60% of WT levels. This showed that the defect of this form of gD on HVEM-expressing cells was primarily one of binding and that this was separable from its later function in virus entry. gD(inverted Delta 243t) showed WT binding affinity for both receptors, but virus containing this form of gD had a markedly reduced rate of entry, suggesting that gD(inverted Delta 243) is impaired in a postbinding step in the entry process. There was no correlation between gD mutant activity in fusion or virus entry and receptor-binding affinity. We conclude that gD functions in virus entry and cell fusion regardless of its receptor-binding kinetics and that as long as binding to a functional receptor occurs, entry will progress. PMID- 12885916 TI - Identification of an insulator in AAVS1, a preferred region for integration of adeno-associated virus DNA. AB - In latent adeno-associated virus (AAV) infection, the viral genome is integrated preferentially into the human chromosome 19 q arm at a specific region designated AAVS1, which has an open chromatin conformation as indicated by the presence of a DNase I-hypersensitive site (DHS-S1). We examined whether an insulator, which defines the domain of gene expression by directionally blocking the action of enhancers and by preventing the spread of heterochomatin, is present near the DHS S1 in the middle of a 2.6-kbp AAVS1-related DNA fragment used in this study. The fragment, cloned into an Epstein-Barr virus (EBV)-based eukaryotic episomal plasmid, was introduced into HEK293 cells. The DHS-S1 on the plasmid replicating in the nuclei was hypersensitive to DNase I digestion, and thus, the EBV plasmid system was used in an enhancer-blocking assay with the 2.6-kbp DNA and two shortened DNAs, of 1.6 kbp and 336 bp, containing DHS-S1. The three DNA fragments, when inserted in the proper direction between the cytomegalovirus immediate-early enhancer and minimal promoter, repressed the expression of a reporter gene. Thus, the enhancer-blocking activity was located within the 336-bp DNA containing the entire region (300 bp) of DHS-S1. To investigate the prevention of repression caused by heterochromatin, a transgene-expressing cassette flanked by the two 336-bp DNAs placed in the enhancer-blocking direction was introduced into HEK293 and HeLa cells. All the cell clones examined with the cassette integrated into cell DNA continued to express the transgene, which indicates that the pair of 336-bp DNA apparently prevented the spread of heterochromatin. The results show that an insulator lies between nucleotides 17 and 354 near the DHS-S1 in AAVS1. In a gel shift test, the 336-bp DNA did not bind an in vitro-prepared CCCTC-binding factor that binds to the chicken beta globin insulator, suggesting that the AAVS1 insulator requires an as yet unidentified binding protein. The newly identified AAVS1 insulator is likely to contribute to the maintenance of an open chromatin conformation that affects the life cycle of AAV. PMID- 12885915 TI - Cellular localization of nectin-1 and glycoprotein D during herpes simplex virus infection. AB - During viral entry, herpes simplex virus (HSV) glycoprotein D (gD) interacts with a specific cellular receptor such as nectin-1 (PRR1/HveC/CD111) or the herpesvirus entry mediator A (HVEM/HveA). Nectin-1 is involved in cell-to-cell adhesion. It is located at adherens junctions, where it bridges cells through homophilic or heterophilic interactions with other nectins. Binding of HSV gD prevents nectin-1-mediated cell aggregation. Since HSV gD affects the natural function of nectin-1, we further investigated the effects of gD expression on nectin-1 during HSV infection or in transfected cells. We also studied the importance of the interaction between nectin-1 and the cytoplasmic protein afadin for HSV entry and spread as well as the effects of infection on this interaction. In these investigations, we used a panel of cells expressing nectin-1 or nectin-1 green fluorescent protein fusions as the only mediators of HSV entry. During HSV infection, nectin-1 localization at adherens junction was dramatically altered in a manner dependent on gD expression. Nectin-1 and gD colocalized at cell contact areas between infected and noninfected cells and at the edges of plaques. This specific accumulation of gD at junctions was driven by expression of nectin-1 in trans on the surface of adjacent cells. Reciprocally, nectin-1 was maintained at junctions by the trans expression of gD in the absence of a cellular natural ligand. Our observations indicate that newly synthesized gD substitutes for nectin-1 of infected cells at junctions with noninfected cells. We propose that gD attracts and maintains the receptor at junctions where it can be used for virus spread. PMID- 12885917 TI - Intracellular trafficking of a palmitoylated membrane-associated protein component of enveloped vaccinia virus. AB - The F13L protein of vaccinia virus, an essential and abundant palmitoylated peripheral membrane component of intra- and extracellular enveloped virions, associates with Golgi, endosomal, and plasma membranes in the presence or absence of other viral proteins. In the present study, the trafficking of a fully functional F13L-green fluorescent protein (GFP) chimera in transfected and productively infected cells was analyzed using specific markers and inhibitors. We found that Sar1(H79G), a trans-dominant-negative protein inhibitor of cargo transport from the endoplasmic reticulum, had no apparent effect on the intracellular distribution of F13L-GFP, suggesting that the initial membrane localization occurs at a downstream compartment of the secretory pathway. Recycling of F13L-GFP from the plasma membrane was demonstrated by partial colocalization with FM4-64, a fluorescent membrane marker of endocytosis. Punctate F13L-GFP fluorescence overlapped with clathrin and Texas red-conjugated transferrin, suggesting that endocytosis occurred via clathrin-coated pits. The inhibitory effects of chlorpromazine and trans-dominant-negative forms of dynamin and Eps15 protein on the recycling of F13L-GFP provided further evidence for clathrin-mediated endocytosis. In addition, the F13L protein was specifically coimmunoprecipitated with alpha-adaptin, a component of the AP-2 complex that interacts with Eps15. Nocodazole and wortmannin perturbed the intracellular trafficking of F13L-GFP, consistent with its entry into late and early endosomes through the secretory and endocytic pathways, respectively. The recycling pathway described here provides a mechanism for the reutilization of the F13L protein following its deposition in the plasma membrane during the exocytosis of enveloped virions. PMID- 12885918 TI - Identification of a C-terminal regulatory motif in hepatitis C virus RNA dependent RNA polymerase: structural and biochemical analysis. AB - The NS5B RNA-dependent RNA polymerase encoded by the hepatitis C virus (HCV) is a key component of the viral replicase. Reported here is the three-dimensional structure of HCV NS5B polymerase, with the highlight on its C-terminal folding, determined by X-ray crystallography at 2.1-A resolution. Structural analysis revealed that a stretch of C-terminal residues of HCV NS5B inserted into the putative RNA binding cleft, where they formed a hydrophobic pocket and interacted with several important structural elements. This region was found to be conserved and unique to the RNA polymerases encoded by HCV and related viruses. Through biochemical analyses, we confirmed that this region interfered with the binding of HCV NS5B to RNA. Deletion of this fragment from HCV NS5B enhanced the RNA synthesis rate up to approximately 50-fold. These results provide not only direct experimental insights into the role of the C-terminal tail of HCV NS5B polymerase but also a working model for the RNA synthesis mechanism employed by HCV and related viruses. PMID- 12885919 TI - Major histocompatibility complex class I alleles associated with slow simian immunodeficiency virus disease progression bind epitopes recognized by dominant acute-phase cytotoxic-T-lymphocyte responses. AB - Certain major histocompatibility complex class I (MHC-I) alleles are associated with delayed disease progression in individuals infected with human immunodeficiency virus (HIV) and in macaques infected with simian immunodeficiency virus (SIV). However, little is known about the influence of these MHC alleles on acute-phase cellular immune responses. Here we follow 51 animals infected with SIV(mac)239 and demonstrate a dramatic association between Mamu-A*01 and -B*17 expression and slowed disease progression. We show that the dominant acute-phase cytotoxic T lymphocyte (CTL) responses in animals expressing these alleles are largely directed against two epitopes restricted by Mamu-A*01 and one epitope restricted by Mamu-B*17. One Mamu-A*01-restricted response (Tat(28-35)SL8) and the Mamu-B*17-restricted response (Nef(165-173)IW9) typically select for viral escape variants in early SIV(mac)239 infection. Interestingly, animals expressing Mamu-A*1 and -B*17 have less variation in the Tat(28-35)SL8 epitope during chronic infection than animals that express only Mamu-A*01. Our results show that MHC-I alleles that are associated with slow progression to AIDS bind epitopes recognized by dominant CTL responses during acute infection and underscore the importance of understanding CTL responses during primary HIV infection. PMID- 12885920 TI - Distinct roles of cellular Lck and p80 proteins in herpesvirus saimiri Tip function on lipid rafts. AB - Lipid rafts are proposed to function as platforms for both receptor signaling and trafficking. Following interaction with antigenic peptides, the T-cell receptor (TCR) rapidly translocates to lipid rafts, where it transmits signals and subsequently undergoes endocytosis. The Tip protein of herpesvirus saimiri (HVS), which is a T-lymphotropic tumor virus, interacts with cellular Lck tyrosine kinase and p80, a WD domain-containing endosomal protein. Interaction of Tip with p80 induces enlarged vesicles and recruits Lck and TCR complex into these vesicles for trafficking. We report here that Tip is constitutively present in lipid rafts and that Tip interaction with p80 but not with Lck is necessary for its efficient localization in lipid rafts. The Tip-Lck interaction was required for recruitment of the TCR complex to lipid rafts, and the Tip-p80 interaction was critical for the aggregation and internalization of lipid rafts. These results suggest the potential mechanism for Tip-mediated TCR downregulation: Tip interacts with Lck to recruit TCR complex to lipid rafts, and it subsequently interacts with p80 to initiate the aggregation and internalization of the lipid raft domain and thereby downregulate the TCR complex. Thus, the signaling and targeting functions of HVS Tip rely on two functionally and genetically separable mechanisms that independently target cellular Lck tyrosine kinase and p80 endosomal protein. PMID- 12885921 TI - Redistribution of cyclophilin A to viral factories during vaccinia virus infection and its incorporation into mature particles. AB - Cyclophilins are peptidyl-prolyl cis-trans isomerases involved in catalyzing conformational changes and accelerating the rate of protein folding and refolding in several cellular systems. In the present study, we analyzed the expression pattern and intracellular distribution of the cellular isomerase cyclophilin A (CypA) during vaccinia virus (VV) infection. An impressive increase in CypA stability was observed, leading to a practically unchanged accumulation of CypA during infection, although its synthesis was completely inhibited at late times. By confocal microscopy, we observed that CypA went through an intense reorganization in the cell cytoplasm and colocalized with the virosomes late in infection. CypA relocation to viral factories required the synthesis of viral postreplicative proteins, and treatment of infected cells with cyclosporine (CsA) prevented CypA relocation, clearly excluding the virosomes from CypA staining. Immunoelectron microscopy of VV-infected cells showed that CypA was incorporated into VV particles during morphogenesis. Biochemical and electron microscopic assays with purified virions confirmed that CypA was encapsidated within the virus particle and localized specifically in the core. This work suggests that CypA may develop an important role in VV replication. PMID- 12885922 TI - Role of the human immunodeficiency virus type 1 envelope gene in viral fitness. AB - A human host offers a variety of microenvironments to the infecting human immunodeficiency virus type 1 (HIV-1), resulting in various selective pressures, most of them directed against the envelope (env) gene. Therefore, it seems evident that the replicative capacity of the virus is largely related to viral entry. In this study we have used growth competition experiments and TaqMan real time PCR detection to measure the fitness of subtype B HIV-1 primary isolates and autologous env-recombinant viruses in order to analyze the contribution of wild type env sequences to overall HIV-1 fitness. A significant correlation was observed between fitness values obtained for wild-type HIV-1 isolates and those for the corresponding env-recombinant viruses (r = 0.93; P = 0.002). Our results suggest that the env gene, which is linked to a myriad of viral characteristics (e.g., entry into the host cell, transmission, coreceptor usage, and tropism), plays a major role in fitness of wild-type HIV-1. In addition, this new recombinant assay may be useful for measuring the contribution of HIV-1 env to fitness in viruses resistant to novel antiretroviral entry inhibitors. PMID- 12885923 TI - Pseudorabies virus US3 protein kinase mediates actin stress fiber breakdown. AB - Disruption of specific components of the host cytoskeleton has been reported for several viruses and is thought to be beneficial for viral replication and spread. Our previous work demonstrated that infection of swine kidney (SK-6) cells with pseudorabies virus (PRV), a swine alphaherpesvirus, induced actin stress fiber breakdown. In the present study, using several PRV deletion mutants, we found that the US3 serine/threonine (S/T) protein kinase is involved in breakdown of actin stress fibers in different PRV-infected cell lines. Further, by transfection assays, we showed that PRV US3 itself, in the absence of other viral proteins, is able to trigger actin stress fiber breakdown when it is localized in sufficient amounts in the nucleus. PMID- 12885924 TI - TT virus loads and lymphocyte subpopulations in children with acute respiratory diseases. AB - TT virus (TTV) produces chronic plasma viremia in around 90% of healthy individuals of all ages and has, therefore, been proposed as a commensal human virus. We recently demonstrated that in children hospitalized for acute respiratory diseases high TTV loads were associated with severe forms of disease. Here, we report that in such children TTV loads showed an inverse correlation with the percentage of circulating total T and helper T cells and a direct correlation with the percentage of B cells. Thus, florid TTV replication might contribute to lymphocyte imbalances and, possibly, immunosuppressive effects, thus resembling related animal viruses. PMID- 12885925 TI - Recombinant avian infectious bronchitis virus expressing a heterologous spike gene demonstrates that the spike protein is a determinant of cell tropism. AB - A recombinant infectious bronchitis virus (IBV), BeauR-M41(S), was generated using our reverse genetics system (R. Casais, V. Thiel, S. G. Siddell, D. Cavanagh, and P. Britton, J. Virol. 75:12359-12369, 2001), in which the ectodomain region of the spike gene from IBV M41-CK replaced the corresponding region of the IBV Beaudette genome. BeauR-M41(S) acquired the same cell tropism phenotype as IBV M41-CK in four different cell types, demonstrating that the IBV spike glycoprotein is a determinant of cell tropism. PMID- 12885926 TI - Expression of immunogenic S1 glycoprotein of infectious bronchitis virus in transgenic potatoes. AB - The expression of infectious bronchitis virus (IBV) S1 glycoprotein in potatoes and its immunogenicity in mice and chickens were investigated. Potato plants were genetically transformed with a cDNA construct encoding the IBV S1 glycoprotein with the Agrobacterium system. Genomic DNA and mRNA analyses of the transformed plantlets confirmed the integration of the foreign cDNA into the potato genome, as well as its transcription. Mice and chickens vaccinated with the expressed IBV S1 glycoprotein produced antibodies that neutralized IBV infectivity. After three immunizations, vaccinated chickens were completely protected from virulent IBV infection. These results demonstrate that transgenic potatoes expressing IBV S1 glycoprotein can be used as a source of recombinant antigen for vaccine production. PMID- 12885927 TI - Vaccinia virus recombinant expressing an 87-kilodalton polyprotein that is sufficient to form astrovirus-like particles. AB - Human astrovirus is an important cause of acute gastroenteritis. We have generated, for the first time, a vaccinia virus recombinant expressing the astrovirus 87-kDa structural polyprotein. The results demonstrate that this expression results in the formation of virus-like particles in the absence of other astrovirus proteins and genomic RNA. The purified trypsin-activated virus like particles strongly resemble the complete astrovirus particles. PMID- 12885928 TI - Novel gamma-1 herpesviruses identified in free-ranging new world monkeys (golden handed tamarin [Saguinus midas], squirrel monkey [Saimiri sciureus], and white faced saki [Pithecia pithecia]) in French Guiana. AB - The recent finding of a novel Epstein-Barr virus-related lymphocryptovirus (CalHV 3) in a captive colony of common marmoset (Callithrix jacchus) in the United States modifies the view that the host range of lymphocryptovirus is restricted to humans and Old World primates. We investigated the presence of Epstein-Barr virus-related viruses in 79 samples of New World monkeys caught in the wild, including six species of the Cebidae family and one of the Callitrichidae, living in the rain forest of French Guiana. Using a degenerate consensus PCR method for the herpesvirus DNA polymerase gene, we identified three novel lymphocryptoviruses from golden-handed tamarin (Saguinus midas) of the Callitrichidae family and squirrel monkey (Saimiri sciureus) and white-faced saki (Pithecia pithecia) of the Cebidae family. With the CalHV-3 strain, these three novel viruses constitute a well-supported phylogenetic clade in the Lymphocryptovirus genus, which is clearly distinct from the lineage of Old World lymphocryptovirus, hosted by catarrhine monkeys and humans. In tamarins, the prevalence of the novel lymphocryptovirus was more than 50%, indicating that it circulates well in the wild population, perhaps due to specific ecoethological patterns such as confrontations and intergroup migration. The detection and partial molecular characterization of the polymerase gene of three novel Gamma-1 Herpesvirinae from New World monkeys caught in the wild clearly indicate that free-ranging populations of platyrrhine are natural hosts of lymphocryptoviruses. Further characterization of these novel viruses will provide new insight not only into the origin and evolution of Gammaherpesvirinae but also into their pathogenicity. PMID- 12885929 TI - Immunity in the eye: lessons in organ-specific responses. AB - Fifty-five years have past since Sir Peter Medawar first demonstrated that the fate of allografts differs in the skin, brain, subcutaneous tissue, and anterior chamber of the eye. Indeed, these and other experiments performed at the University of Birmingham and University College London not only helped define key paradigms in transplantation biology but introduced the concept of immune privilege in the eye and in other tissues. In the ensuing years, the work from dozens of laboratories has confirmed that immune responses in eye, although sharing many features with immunity in other tissues, are atypical in many respects. This has been hypothesized to stem from the requirement in the eye to hold inflammation in check to preserve visual function. Indeed, inflammatory reactions in different regions of the eye can result in visual impairment and blindness, manifesting in diseases such as vernal keratoconjunctivitis, uveitis, corneal endotheliitis, Mooren's ulcer, and pemphigoid. The investigation of the cell and molecular basis of the unique nature of immunity in the eye has revealed a remarkably complex network of interactions that controls inflammation while affording immunity. The most recent data generated in this field and the concepts arising from their interpretation were discussed at a symposium held at the University College London in late 2002. Here, we highlight some of the findings discussed at the meeting and introduce in-depth reviews published in this issue of the journal. PMID- 12885930 TI - Retinal transplantation: progress and problems in clinical application. AB - There is currently no real treatment for blinding disorders that stem from the degeneration of cells in the retina and affect at least 50 million individuals worldwide. The excitement that accompanied the first studies showing the potential of retinal cell transplantation to alleviate the progress of blindness in such diseases as retinitis pigmentosa and age-related macular degeneration has lost some of its momentum, as attempts to apply research to the clinic have failed so far to provide effective treatments. What these studies have shown, however, is not that the approach is flawed but rather that the steps that need to be taken to achieve a viable, clinical treatment are many. This review summarizes the course of retinal transplant studies and points to obstacles that still need to be overcome to improve graft survival and efficacy and to develop a protocol that is effective in a clinical setting. Emphasis is given particularly to the consequences of introducing transplants to sites that have been considered immunologically privileged and to the role of the major histocompatibility complex classes I and II molecules in graft survival and rejection. PMID- 12885931 TI - Control of myeloid activity during retinal inflammation. AB - Combating myeloid cell-mediated destruction of the retina during inflammation or neurodegeneration is dependent on the integrity of homeostatic mechanisms within the tissue that may suppress T cell activation and their subsequent cytokine responses, modulate infiltrating macrophage activation, and facilitate healthy tissue repair. Success is dependent on response of the resident myeloid-cell populations [microglia (MG)] to activation signals, commonly cytokines, and the control of infiltrating macrophage activation during inflammation, both of which appear highly programmed in normal and inflamed retina. The evidence that tissue CD200 constitutively provides down-regulatory signals to myeloid-derived cells via cognate CD200-CD200 receptor (R) interaction supports inherent tissue control of myeloid cell activation. In the retina, there is extensive neuronal and endothelial expression of CD200. Retinal MG in CD200 knockout mice display normal morphology but unlike the wild-type mice, are present in increased numbers and express nitric oxide synthase 2, a macrophage activation marker, inferring that loss of CD200 or absent CD200R ligation results in "classical" activation of myeloid cells. Thus, when mice lack CD200, they show increased susceptibility to and accelerated onset of tissue-specific autoimmunity. PMID- 12885932 TI - The immune privilege of corneal grafts. AB - Keratoplasty is the oldest and one of the most successful forms of solid tissue transplantation. In the United States, over 33,000 corneal transplants are performed each year. Unlike other forms of tissue transplantation, keratoplasties are routinely performed without the aid of tissue typing or systemic immunosuppressive drugs. In spite of this, 90% of the first-time corneal transplants will succeed-a condition that demonstrates the immune privilege of keratoplasties. The avascular nature of the corneal allograft bed led many to suspect that corneal grafts were sequestered from the immune apparatus. Although pleasing in its simplicity, this explanation has given way to a more comprehensive hypothesis that embodies multiple, interdependent mechanisms, which promote the long-term survival of corneal allografts. These mechanisms conspire to interrupt the transmission of immunogenic stimuli to peripheral lymphoid tissues; induce the generation of a deviated immune response; and neutralize immune effector elements at the host-graft interface. This paradigm is analogous to a three-legged stool. Disassembly of any one of the three components results in the collapse of immune privilege. Strategies to re-establish corneal immune privilege may have clinical application for high-risk hosts who have rejected previous corneal allografts. PMID- 12885933 TI - Corneal immunity is mediated by heterogeneous population of antigen-presenting cells. AB - Corneal antigen-presenting cells (APC), including dendritic cells (DC), were thought to reside exclusively in the peripheral cornea. Here, we present recent data from our group demonstrating that the central cornea is indeed endowed with a heterogeneous population of epithelial and stromal DC, which function as APC. Although the corneal periphery contains mature and immature resident bone marrow derived CD11c(+) DC, the central cornea is endowed exclusively with immature and precursor DC, both in the epithelium and the stroma, wherein Langerhans cells and monocytic DC reside, respectively. During inflammation, a majority of resident DC undergo maturation by overexpressing major histocompatibility complex class II and B7 (CD80/CD86) costimulatory molecules. In addition to the DC, macrophages are present in the posterior corneal stroma. In transplantation, donor-derived DC are able to migrate to host cervical lymph nodes and activate host T cells via the direct pathway when allografts are placed in inflamed host beds. These data revise the tenet that the cornea is immune-privileged as a result of lack of resident lymphoreticular cells and suggest that the cornea is capable of diverse cellular mechanisms for antigen presentation. PMID- 12885934 TI - Ocular immune privilege: the eye takes a dim but practical view of immunity and inflammation. AB - The delicate visual axis that makes precise vision possible is highly vulnerable to the destructive potential of immunogenic inflammation. Immune privilege of the eye is the experimental expression of the way in which evolution has coped with the countermanding threats to vision of ocular infections and ocular immunity and inflammation. Ocular immune privilege has five primary features that account for its existence: blood:ocular barriers, absent lymphatic drainage pathways, soluble immunomodulatory factors in aqueous humor, immunomodulatory ligands on the surface of ocular parenchymal cells, and indigenous, tolerance-promoting antigen presenting cells (APCs). Three manifestations of ocular immune privilege that have received the most extensive study are the intraocular microenvironment, which is selectively anti-inflammatory and immunosuppressive; the prolonged acceptance of solid tissue and tumor allografts in the anterior chamber; and the induction of systemic tolerance to eye-derived antigens. Anterior chamber associated immune deviation is known to arise when indigenous, ocular APCs capture eye-derived antigens and deliver them to the spleen where multicellular clusters of these cells, natural killer T cells, marginal zone B cells, and gammadelta T cells create an antigen-presentation environment that leads to CD4(+) and CD8(+) alpha/beta T cells, which as regulators, suppress induction and expression of T helper cell type 1 (Th1) and Th2 immune expression systems. The ways the eye influences local and systemic immune responses to ocular antigens and pathogens carry risks to and benefits for mammalian organisms. As loss of sight is a powerful, negative-selecting force, the benefits of ocular immune privilege outweigh the risks. PMID- 12885935 TI - Inhibition of myeloid cell differentiation in cancer: the role of reactive oxygen species. AB - It is well established that tumor growth is associated with accumulation of immature myeloid cells (ImC). They play an important role in tumor-associated immune suppression. ImC accumulate not only in tumor-bearing hosts but also in immunized, tumor-free hosts or hosts infected with bacterial pathogens. The kinetics of ImC in these mice is different. If in tumor-bearing mice, the number of ImC continues to increase with tumor progression in tumor-free mice after an initial spike, it decreases to the control level. Here, we have investigated the mechanisms of ImC accumulation in tumor-bearing hosts by comparing differentiation of ImC obtained from tumor-free and tumor-bearing mice. In the presence of appropriate growth factors, ImC isolated from tumor-free mice quickly differentiated in vitro into mature dendritic cells (DC), macrophages, and granulocytes. In contrast, differentiation of ImC from tumor-bearing mice was significantly delayed. Similar results were obtained in vivo after adoptive transfer of ImC into naive, congeneic mice. ImC transferred into tumor-bearing recipients failed to differentiate into DC or macrophages. ImC from tumor-bearing mice had significantly higher levels of reactive oxygen species (ROS) than ImC obtained from tumor-free mice. Hydrogen peroxide (H(2)O(2)) but not superoxide radical anions was found to be the major part of this increased ROS production. In vitro experiments demonstrated that scavenging of H(2)O(2) with catalase induced differentiation of ImC from tumor-bearing mice into macrophages. Thus, this is a first demonstration that tumors may prevent differentiation of antigen presenting cells by increasing the level of endogenous H(2)O(2) in immature myeloid cells. PMID- 12885936 TI - Hypoxia reduces CD80 expression on monocytes but enhances their LPS-stimulated TNF-alpha secretion. AB - Monocytes/macrophages in ischemic tissues are involved in inflammation and suppression of adaptive immunity via secretion of proinflammatory cytokines and reduced ability to trigger T cells, respectively. We subjected human mononuclear cells and mouse macrophages to hypoxia and reoxygenation, the main constituents of ischemia and reperfusion, and added lipopolysaccharide (LPS) to simulate bacterial translocation, which frequently accompanies ischemia. We monitored the secretion of tumor necrosis factor alpha (TNF-alpha) and the surface expression of human leukocyte antigen-DR and the costimulatory molecules CD80 and CD86 on monocytes/macrophages. Hypoxia selectively reduced the surface expression of CD80 (P<0.01), and synergistically with LPS, it enhanced TNF-alpha secretion (P<0.003). Reoxygenation reversed both phenomena. In the mouse macrophage cell line RAW 264.7, hypoxia reduced the surface expression of CD80 and increased its concentrations in the supernatants (P<0.01). Down-regulation of the mRNA coding for the membrane-anchored CD80 was observed, suggesting that hypoxia triggers alternative splicing to generate soluble CD80. Cumulatively, these results suggest that hypoxia simultaneously affects monocytes/macrophages to enhance inflammation and reduce their ability to initiate adaptive-immunity responses associated with ischemic injury. PMID- 12885937 TI - Mannose receptor contribution to Candida albicans phagocytosis by murine E-clone J774 macrophages. AB - Mannoproteins, as the main constituents of the outer layer of yeast cell walls, are able to interact with phagocytic cells in an opsonin-independent manner through the mannose receptor (MR) and to induce yeast ingestion by the professional phagocytes. Moreover, the MR also mediates endocytosis of soluble ligands through clathrin-coated pits. Here, we studied some aspects of the interaction between the MR and Candida albicans using murine E-clone macrophages and the consequences on MR trafficking. Using a pull-down assay involving mixture E-clone macrophage detergent lysate with mannosylated Sepharose beads and glutaraldehyde-fixed, heat-killed (HK) C. albicans, we found that binding of solubilized MR to mannosylated particles occurred with characteristics similar to the receptor's cell-surface mannose-binding activity. We then demonstrated that MR expressed on E-clone macrophages contributed to phagocytosis of unopsonized, HK C. albicans and that yeast phagocytosis induced a decrease in MR endocytic activity without concomitant degradation of the receptor in the time lapse studied. PMID- 12885938 TI - Analysis of the maturation process of dendritic cells deficient for TNF and lymphotoxin-alpha reveals an essential role for TNF. AB - Dendritic cells (DCs) generated from bone marrow (BM) precursor cells of C57BL/6 (B6.WT) mice and cultured in the presence of granulocyte macrophage-colony stimulating factor differentiate to mature BM-DCs spontaneously. These mature DCs are characterized by high levels of major histocompatibility complex (MHC) class II, CD40, and CD86 on their surface. To analyze the involvement of tumor necrosis factor (TNF) and the related cytokine lymphotoxin (LT)alpha in DC maturation, we studied the development of DCs from the BM of B6.TNF(-/-), B6.LTalpha(-/-), and B6.TNF/LTalpha(-/-) mice and compared it to B6.WT mice. Although the development of BM precursor cells to the level of immature DCs (CD11c(+), MHC class II(low), CD40(low), and CD86(low)) was equivalent in all genotypes, B6.TNF(-/-) and B6.TNF/LTalpha(-/-) cells showed an impaired capacity to differentiate to mature DCs. In contrast, mature BM-DCs generated from LTalpha-negative, immature DCs developed like B6.WT cells. Further studies revealed that once matured, the phenotype of all tested genotypes was comparable. They expressed high levels of CD40 and CD86, were exclusively positive for the chemokine receptor (CCR)7 but negative for CCR5 and CCR2, and were able to enter the paracortex of draining lymph nodes. The limited maturation of TNF-deficient BM-DCs could be restored by mixing TNF-negative with TNF-positive Ly5.1 BM cells, and maturation of B6.WT DCs could be blocked with an anti-TNF monoclonal antibody. The substitution of B6.TNF(-/-) BM cells with recombinant TNF revealed promotion or suppression of BM DC maturation depending on the point of time of TNF addition. PMID- 12885939 TI - Tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) and TNF alpha promote the NF-kappaB-dependent maturation of normal and leukemic myeloid cells. AB - Tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) and TNF alpha induced monocytic maturation of primary normal CD34-derived myeloid precursors and of the M2/M3-type acute myeloid leukemia HL-60 cell line, associated to increased nuclear factor (NF)-kappaB activity and nuclear translocation of p75, p65, and p50 NF-kappaB family members. Consistently, both cytokines also induced the degradation of the NF-kappaB inhibitors, IkappaBalpha and IkappaB epsilon, and up-regulated the surface expression of TRAIL-R3, a known NF-kappaB target. However, NF-kappaB activation and IkappaB degradation occurred with different time-courses, since TNF-alpha was more potent, rapid, and transient than TRAIL. Of the two TRAIL receptors constitutively expressed by HL 60 (TRAIL-R1 and TRAIL-R2), only the former was involved in IkappaB degradation, as demonstrated by using agonistic anti-TRAIL receptor antibodies. Moreover, NF kappaB nuclear translocation induced by TRAIL but not by TNF-alpha was abrogated by z-IETD-fmk, a caspase-8-specific inhibitor. The key role of NF-kappaB in mediating the biological effects of TNF-alpha and TRAIL was demonstrated by the ability of unrelated pharmacological inhibitors of the NF-kappaB pathway (parthenolide and MG-132) to abrogate TNF-alpha- and TRAIL-induced monocytic maturation. These findings demonstrate that NF-kappaB is essential for monocytic maturation and is activated via distinct pathways, involving or not involving caspases, by the related cytokines TRAIL and TNF-alpha. PMID- 12885940 TI - Cytokine requirements for the growth and development of mouse NK cells in vitro. AB - Natural killer (NK) cells arise from immature progenitors present in fetal tissues and adult bone marrow, but the factors responsible for driving the proliferation and differentiation of these progenitors are poorly understood. Mouse NK cells had previously been thought not to express interleukin (IL) 2Ralpha chains, but we show here that immature and mature mouse NK cells express IL-2Ralpha chain mRNA and that low levels of IL-2Ralpha chains can be detected on the surface of immature and mature NK cells provided they are cultured in the absence of IL-2. Despite their potential expression of high-affinity IL-2 receptors, immature NK cells only proliferate if IL-2 is present at extremely high concentrations. Surprisingly, IL-15 can also only support the growth of immature NK cells at high, presumably nonphysiological concentrations. Although NK cells express mRNA for the high-affinity IL-15Ralpha chain, they also express a variety of alternately spliced transcripts whose protein products could potentially disrupt signaling through IL-15 receptors. The requirement for high concentrations of IL-2 and IL-15 suggests that if these cytokines play any role in the proliferative expansion of NK cells in vivo, they act indirectly via other cells or in cooperation with other factors. In support of the latter possibility, we report that the recently described cytokine IL-21 can markedly enhance the proliferation of immature (and mature) NK cells in the presence of doses of IL-2 and IL-15 that by themselves have little growth-promoting activity. PMID- 12885941 TI - Expression of L-CCR in HEK 293 cells reveals functional responses to CCL2, CCL5, CCL7, and CCL8. AB - It has become clear in the past years that chemokines and chemokine receptors are pivotal regulators of cellular communication and trafficking. In addition to the approximately 20 chemokine receptors that have been cloned and described, various orphan receptors with a chemokine receptor-like structure are known. We have investigated the orphan mouse chemokine receptor (L-CCR) in HEK 293 cells, a receptor that was originally described in a mouse macrophage cell line. Cells expressing this receptor show pertussis toxin-sensitive chemotaxis and small intracellular calcium transients in response to the chemokines CCL2, CCL7, CCL8, and CCL5. Biotinylated CCL2 binds to L-CCR-expressing cells, and transfection experiments with an L-CCR-green fluorescent protein fusion protein showed L-CCR expression in the membranes of recombinant HEK 293 cells. Although radioligand binding was not detected, it is suggested that L-CCR is a functional chemokine receptor. PMID- 12885942 TI - Chemokine receptor expression and chemotactic responsiveness of human monocytes after influenza A virus infection. AB - Chemokines and their receptors play an important role in site-directed migration and activation of leukocytes. To understand how viral infections may impair this function, we analyzed chemokine receptor expression and responsiveness of human monocytes after infection with influenza A virus. Whereas treatment with infectious virus induced a rapid down-regulation of the CCL2/monocyte chemoattractant protein-1 (MCP-1)-specific receptor CCR2, inactivated virus did not significantly alter CCR2 surface expression. In parallel, the response to CCL2/MCP-1 was lost after infection with active virus: Neither a CCL2/MCP-1 induced shift of intracellular calcium concentrations nor the chemotactic response to CCL2/MCP-1 was detectable. In striking contrast, the presence of CCR1 and CCR5 on the cell surface remained unchanged or was even slightly up-regulated after viral infection. However, the remaining expression of CCR1 and CCR5 correlated reciprocally with an ongoing unresponsiveness to the CCR1 and CCR5 agonists CCL3/macrophage-inflammatory protein-1alpha (MIP-1alpha), CCL4/MIP 1beta, and CCL5/regulated on activation, normal T expressed and secreted (RANTES), all chemokines binding to these two receptors. The CCL3/MIP-1alpha induced shifts of intracellular calcium concentrations declined gradually to almost undetectable levels, and most conspiciuously, the chemotactic response to CCL3/MIP-1alpha, CCL4/MIP-1beta, and CCL5/RANTES was lost after infection with active influenza virus. Inactivated virus particles did not significantly alter the responsiveness induced by CCR1 and CCR5 agonists. Despite the inability of chemokine receptors to elicit migration, phosphorylation of protein kinase B was not altered in virus-infected monocytes. Thus, influenza A virus infection rapidly abolishes the functional responsiveness of monocytes and prevents an adequate response of the infected cells to chemokine stimulation. PMID- 12885943 TI - A role for Syk-kinase in the control of the binding cycle of the beta2 integrins (CD11/CD18) in human polymorphonuclear neutrophils. AB - A fine control of beta(2) integrin (CD11/CD18)-mediated firm adhesion of human neutrophils to the endothelial cell monolayer is required to allow ordered emigration. To elucidate the molecular mechanisms that control this process, intracellular protein tyrosine signaling subsequent to beta(2) integrin-mediated ligand binding was studied by immunoprecipitation and Western blotting techniques. The 72-kDa Syk-kinase, which was tyrosine-phosphorylated upon adhesion, was found to coprecipitate with CD18, the beta-subunit of the beta(2) integrins. Moreover, inhibition of Syk-kinase by piceatannol enhanced adhesion and spreading but diminished N-formyl-Met-Leu-Phe-induced chemotactic migration. The enhancement of adhesiveness was associated with integrin clustering, which results in increased integrin avidity. In contrast, piceatannol had no effect on the surface expression or on the affinity of beta(2) integrins. Altogether, this suggests that Syk-kinase controls alternation of beta(2) integrin-mediated ligand binding with integrin detachment. PMID- 12885944 TI - Dramatic increase of telomerase activity during dendritic cell differentiation and maturation. AB - Telomerase, the reverse transcriptase that maintains telomere DNA, is usually undetectable in most adult tissues but is positive in embryonic tissues and in cancers. In addition, freshly islolated or in vitro-activated lymphocytes were shown to express high levels of telomerase activity, although its expression in myeloid cells including dendritic cells (DCs) is largely unknown. Here, we investigated telomerase activity during the differentiation and maturation process of DCs. In vitro culture of bone marrow (BM) cells with granulocyte macrophage-colony stimulating factor and interleukin-4 induced a dramatic increase of telomerase activity accompanied with their differentiation into DCs. Furthermore, stimulation with microbial components such as lipopolysaccharide (LPS), which triggers maturation of DCs, augmented the activity. In vivo responses of telomerase activity were also observed in splenic DCs by injection of LPS intraperitoneally. It is interesting that in old mice, telomerase activity of splenic DCs was significantly higher than young mice but rather decreased after LPS stimulation. By measuring expression of cell-surface activation markers, splenic DCs of old mice responded poorly to LPS stimulation. Such poor responses to LPS were also observed in BM-derived DCs. These different features of DCs between young and old mice may contribute to a pathogenesis to microbial infections. PMID- 12885945 TI - TLR2 and TLR4 serve distinct roles in the host immune response against Mycobacterium bovis BCG. AB - Toll-like receptor (TLR) proteins mediate cellular activation by microbes and microbial products. To delineate the role of TLR proteins in the development of host immune responses against mycobacteria, wild-type and TLR-deficient mice were infected with nonpathogenic Mycobacterium bovis bacillus Calmette-Guerin (BCG). Two weeks after intraperitoneal challenge with BCG, few bacilli were present in the lungs of wild-type and TLR4(-/-) mice, whereas bacterial loads were tenfold higher in the lungs of infected TLR2(-/-) mice. BCG challenge in vitro strongly induced proinflammatory cytokine secretion by macrophages from wild-type and TLR4(-/-) mice but not by TLR2(-/-) macrophages. In contrast, intracellular uptake, intracellular bacterial growth, and suppression of intracellular bacterial growth in vitro by interferon-gamma (IFN-gamma) were similar in macrophages from all three mouse strains, suggesting that BCG growth in the lungs of TLR2(-/-) mice was a consequence of defective adaptive immunity. Antigenic stimulation of splenocytes from infected wild-type and TLR4(-/-) mice induced T cell proliferation in vitro, whereas T cells from TLR2(-/-) mice failed to proliferate. Unexpectedly, activated CD4(+) T cells from both TLR-deficient mouse strains secreted little IFN-gamma in vitro compared with control T cells. A role for TLR4 in the control of bacterial growth and IFN-gamma production in vivo was observed only when mice were infected with higher numbers of BCG. Thus, TLR2 and TLR4 appear to regulate distinct aspects of the host immune response against BCG. PMID- 12885946 TI - Pathways for the regulation of interferon-gamma-inducible genes by iron in human monocytic cells. AB - To elucidate iron-regulated interferon-gamma (IFN-gamma) effector functions, we investigated three IFN-gamma-inducible genes [intercellular adhesion molecule-1 (ICAM-1), human leukocyte antigen (HLA)-DR, guanosine 5'-triphosphate cyclohydrolase I (GTP-CH)] in primary human monocytes and the cell line THP-1. IFN-gamma increased the surface expression of ICAM-1 and HLA-DR and stimulated GTP-CH activity. Addition of iron before cytokine stimulation resulted in a dose dependent reduction of these pathways, and iron restriction by desferrioxamine (DFO) enhanced ICAM-1, HLA-DR, and GTP-CH expression. Iron neither affected IFN gamma binding to its receptor nor IFN-gamma receptor surface expression. IFN gamma-inducible mRNA expression of ICAM-1, HLA-DR, and GTP-CH was reduced by iron and increased by DFO by a transcriptional mechanism. Moreover, ICAM-1 and to a lesser extent, GTP-CH and HLA-DR mRNA expression were regulated post transcriptionally, as iron pretreatment resulted in shortening the mRNA half-life compared with cells treated with IFN-gamma alone. Thus, iron perturbations regulate IFN-gamma effector pathways by transcriptional and post-transcriptional mechanisms, indicating that iron rather interferes with IFN-gamma signal transduction processes. PMID- 12885947 TI - High expression of p21 Waf1 in sarcoid granulomas: a putative role for long lasting inflammation. AB - In sarcoid granulomas, apoptotic events are reduced, which explains their characteristic long-lasting inflammation. We have described that interferon-gamma (IFN-gamma) inhibits apoptosis in macrophages through the expression of p21(Waf1). Here, we explore the molecular mechanisms involved in the inhibition of apoptosis in sarcoid granulomas. We analyzed skin biopsies from 19 sarcoidosis patients and 16 controls. Total RNA was subjected to semiquantitative reverse transcriptase-polymerase chain reaction analysis. There was no difference found in the expression of proapoptotic (Bax and Bcl-X(s)) or antiapoptotic (Bcl-2 and Bcl-X(L)) genes nor in the expression of the tumor suppressor gene p53. Furthermore, the expression of IFN-gamma and the cdk inhibitors p21(Waf1) and p27(Kip1) were analyzed. IFN-gamma was detected in 37% of the sarcoidosis patients, and controls were negative (P<0.02). In addition, a higher proportion of patients expressing p21(Waf1) (58%) versus controls (12%) was found (P<0.005). There was a significant correlation between the expression of IFN-gamma and p21(Waf1) (r=0.69) and between p21(Waf1) and fibronectin (r=0.65). Finally, using immunohistochemistry, high p21(Waf1) reactivity was observed inside the granuloma. We conclude that the high levels of p21(Waf1) in sarcoidosis may explain the absence of apoptosis in the granuloma and the persistence of inflammation. PMID- 12885948 TI - DLX genes as targets of ALL-1: DLX 2,3,4 down-regulation in t(4;11) acute lymphoblastic leukemias. AB - Dlx genes constitute a gene family thought to be essential in morphogenesis and development. We show here that in vertebrate cells, Dlx genes appear to be part of a regulatory cascade initiated by acute lymphoblastic leukemia (ALL)-1, a master regulator gene whose disruption is implicated in several human acute leukemias. The expression of Dlx2, Dlx3, Dlx5, Dlx6, and Dlx7 was absent in All-1 -/- mouse embryonic stem cells and reduced in All-1 +/- cells. In leukemic patients affected by the t(4;11)(q21;q23) chromosomal abnormality, the expression of DLX2, DLX3, and DLX4 was virtually abrogated. Our data indicate that Dlx genes are downstream targets of ALL-1 and could be considered as important tools for the study of the early leukemic cell phenotype. PMID- 12885951 TI - Comparative proteomic analysis of proliferating and functionally differentiated mammary epithelial cells. AB - Proliferation and differentiation of mammary epithelial cells are governed by hormonal stimuli, cell-cell, and cell-matrix interactions. Terminal differentiation of mammary epithelial cells depends upon the action of the lactogenic hormones, insulin, glucocorticoids, and prolactin that enable them to synthesize and secrete milk proteins. These differentiated cells are polarized and carry out vectorial transport of milk constituents across the apical plasma membrane. To gain additional insights into the mechanisms governing differentiation of mammary epithelial cells, we identified proteins whose expression distinguishes proliferating from differentiated mammary epithelial cells. For this purpose we made use of the HC11 mammary epithelial line, which is capable of differentiation in response to lactogenic hormones. Using two dimensional gel electrophoresis and mass spectrometry, we found about 60 proteins whose expression levels changed in between these two differentiation states. Bioinformatic analysis revealed differential expression of cytoskeletal components, molecular chaperones and regulators of protein folding and stability, calcium-binding proteins, and components of RNA-processing pathways. The actin cytoskeleton is asymmetrically distributed in differentiated epithelial cells, and the identification of proteins involved in mRNA binding and localization suggests that asymmetry might in part be achieved by controlling cellular localization of mRNAs. The proteins identified provide insights into the differentiation of mammary epithelial cells and the regulation of this process. PMID- 12885952 TI - Identification of novel phosphorylation sites on Xenopus laevis Aurora A and analysis of phosphopeptide enrichment by immobilized metal-affinity chromatography. AB - Mass spectrometric analysis of proteolytically derived phosphopeptides has developed into a widespread technique for the identification of phosphorylated amino acids. Using liquid chromatography-electrospray ionization tandem mass spectrometry, 14 phosphorylation sites were identified on Xenopus laevis His6 Aurora A, a highly conserved regulator of centrosome maturation and cell division. These included seven novel phosphorylation sites, Ser-12, Thr-21, Thr 103, Ser-116, Thr-122, Tyr-155, and Thr-294, as well as the previously identified regulatory sites, Ser-53, Thr-295, and Ser-349. The identification of these novel phosphorylation sites will be important for future studies aimed at elucidating the mechanisms of Aurora A regulation by phosphorylation. Furthermore, we demonstrate that a "kinase-inactive" mutant of Aurora A, K169R, still retains 10% of activity of the wild-type enzyme in vitro along with occupancy of Thr-295 and Ser-12. However, mutation of Asp-281 to Ala completely abolishes activity of the enzyme and should therefore be used preferentially as a genuine kinase-dead construct. Because of the abundance of phosphorylated residues on His6-Aurora A, we found this protein to be an ideal tool for the characterization of immobilized metal-affinity chromatography (IMAC) as a method for phosphopeptide enrichment from complex mixtures. We present a detailed analysis of the binding and elution properties of both the phosphopeptides and unphosphorylated peptides of His6 Aurora A to Fe3+-IMAC before and after methyl esterification. Moreover, we demonstrate a significant difference in enrichment of phosphopeptides when different resins are used for Fe3+-IMAC and characterize the strengths and limitations of this methodology for the study of phosphoproteomics. PMID- 12885953 TI - Evolution of the TCP gene family in Asteridae: cladistic and network approaches to understanding regulatory gene family diversification and its impact on morphological evolution. AB - In the plant subclass Asteridae, bilaterally symmetrical flowers have evolved from a radially symmetrical ancestral phenotype on at least three independent occasions: in the Boraginaceae, Solanaceae, and Lamiales. Development of bilateral flower symmetry has been shown to be determined by the early-acting cycloidea (cyc) and dichotoma (dich) genes in Antirrhinum, a member of the Lamiales. cyc and dich belong to the TCP gene family of putative transcription factors. TCP gene sequences were isolated from 11 Asteridae taxa using an array of degenerate PCR primers. Closely related species exhibiting either ancestral actinomorphic or derived zygomorphic flowers were sampled for each independent origin of bilateral flower symmetry. Cladistic and network-based analyses were performed to establish viable hypotheses regarding the evolution of bilateral symmetry in Asteridae. For the TCP gene family, the use of cladistic phylogenetic analysis to identify orthologous genes is complicated by a paucity of alignable data, frequent gene duplication and extinction, and the possibility of reticulate evolution via intergenic recombination. These complicating factors can be generalized to many regulatory gene families. As an alternative to cladistic analysis, we propose the use of network analysis for the reconstruction of regulatory gene family phylogenetic and functional relationships. Results of analyses support the hypothesis that the origin of bilaterally symmetrical flowers in the Boraginaceae and Solanaceae did not require orthologs or functional analogs of cyc or dich. This suggests that the genetic mechanism that determines bilateral flower symmetry in these taxa is not homologous to that of the Lamiales. Results of analyses are consistent with the hypothesis that the evolution of bilateral floral symmetry in the Lamiales required the origin of a novel gene function subsequent to gene duplication. PMID- 12885954 TI - Variability and phylogenetic incongruence of an SSU nrDNA group I intron in Artomyces, Auriscalpium, and Lentinellus (Auriscalpiaceae: Homobasidiomycetes). AB - Previous research has shown that a group I intron occurs in the SSU ribosomal DNA gene of isolates of Artomyces (Clavicorona, in part) and Lentinellus, but apparently it is absent in an Auriscalpium isolate. However, further investigation revealed that the intron is apparently absent in some species of Artomyces and Lentinellus and is present in at least one species of Auriscalpium. To examine this further, the presence or absence of the group I intron is reported for 13 species of Lentinellus, two species of Auriscalpium, and 16 species of Artomyces. The presence of the intron among the species was variable and is documented for seven species of Lentinellus, one species of Auriscalpium, and 12 species of Artomyces. Furthermore, the presence of the intron was variable among the isolates of several species, and variability of its presence was observed within single isolates, indicating inter-ribosomal repeat heterogeneity. Independent phylogenetic estimations were generated for the intron and nuclear ribosomal internal transcribed spacer regions (ITS). Tests of congruence for the two trees indicated that the data were heterogeneous. Some of the discontinuity between the two phylogenies is due to placement of the Ar. austropiperatus intron within the Lentinellus intron clade. Variability in the length of the intron was observed in populations of the pan-Northern Temperate species Ar. pyxidatus. This was due to the presence of an additional unknown insertional element found only within North American collections of Ar. pyxidatus and absent from European and Asian collections. PMID- 12885955 TI - Patterns of variation within self-incompatibility loci. AB - Diverse self-incompatibility (SI) mechanisms permit flowering plants to inhibit fertilization by pollen that express specificities in common with the pistil. Characteristic of at least two model systems is greatly reduced recombination across large genomic tracts surrounding the S-locus, which regulates SI. In three angiosperm families, including the Solanaceae, the gene that controls the expression of gametophytic SI in the pistil encodes a ribonuclease (S-RNase). The gene that controls pollen SI expression is currently unknown, although several candidates have recently been proposed. Although each candidate shows a high level of polymorphism and complete allelic disequilibrium with the S-RNase gene, such properties may merely reflect tight linkage to the S-locus, irrespective of any functional role in SI. We analyzed the magnitude and nature of nucleotide variation, with the objective of distinguishing likely candidates for regulators of SI from other genes embedded in the S-locus region. We studied the S-RNase gene of the Solanaceae and 48A, a candidate for the pollen gene in this system, and we also conducted a parallel analysis of the regulators of sporophytic SI in Brassica, a system in which both the pistil and pollen genes are known. Although the pattern of variation shown by the pollen gene of the Brassica system is consistent with its role as a determinant of pollen specificity, that of 48A departs from expectation. Our analysis further suggests that recombination between 48A and S-RNase may have occurred during the interval spanned by the gene genealogy, another indication that 48A may not regulate SI expression in pollen. PMID- 12885956 TI - Functional antifreeze glycoprotein genes in temperate-water New Zealand nototheniid fish infer an Antarctic evolutionary origin. AB - The fish fauna of the Antarctic Ocean is dominated by five endemic families of the Perciform suborder Notothenioidei, thought to have arisen in situ within the Antarctic through adaptive radiation of an ancestral stock that evolved antifreeze glycoproteins (AFGPs) enabling survival as the ocean chilled to subzero temperatures. The endemism results from geographic confinement imposed by a massive oceanographic barrier, the Antarctic Circumpolar Current, which also thermally isolated Antarctica over geologic time, leading to its current frigid condition. Despite this voluminous barrier to fish dispersal, a number of species from the Antarctic family Nototheniidae now inhabit the nonfreezing cool temperate coasts of the southern continents. The origin of these temperate-water nototheniids is not completely understood. Since the AFGP gene apparently evolved only once, before the Antarctic notothenioid radiation, the presence of AFGP genes in extant temperate-water nototheniids can be used to infer an Antarctic evolutionary origin. Genomic Southern analysis, PCR amplification of AFGP genes, and sequencing showed that Notothenia angustata and Notothenia microlepidota endemic to southern New Zealand have two to three AFGP genes, structurally the same as those of the Antarctic nototheniids. At least one of these genes is still functional, as AFGP cDNAs were obtained and low levels of mature AFGPs were detected in the blood. A phylogenetic tree based on complete ND2 coding sequences showed monophyly of these two New Zealand nototheniids and their inclusion in the monophyletic Nototheniidae consisted of mostly AFGP-bearing taxa. These analyses support an Antarctic ancestry for the New Zealand nototheniids. A divergence time of approximately 11 Myr was estimated for the two New Zealand nototheniids, approximating the upper Miocene northern advance of the Antarctic Convergence over New Zealand, which might have served as the vicariant event that lead to the northward dispersal of their most recent common ancestor. Similar secondary northward dispersal likely applies to the South American nototheniid Paranotothenia magellanica, which has four AFGP genes in its DNA, but not to the sympatric nototheniid Patagonotothen tessellata, which does not appear to have any AFGP sequences in its genome at all. PMID- 12885957 TI - Early vertebrate evolution of the TATA-binding protein, TBP. AB - TBP functions in transcription initiation in all eukaryotes and in Archaebacteria. Although the 181-amino acid (aa) carboxyl (C-) terminal core of the protein is highly conserved, TBP proteins from different phyla exhibit diverse sequences in their amino (N-) terminal region. In mice, the TBP N terminus plays a role in protecting the placenta from maternal rejection; however the presence of similar TBP N-termini in nontherian tetrapods suggests that this domain also has more primitive functions. To gain insights into the pretherian functions of the N-terminus, we investigated its phylogenetic distribution. TBP cDNAs were isolated from representative nontetrapod jawed vertebrates (zebrafish and shark), from more primitive jawless vertebrates (lamprey and hagfish), and from a prevertebrate cephalochordate (amphioxus). Results showed that the tetrapod N-terminus likely arose coincident with the earliest vertebrates. The primary structures of vertebrate N-termini indicates that, historically, this domain has undergone events involving intragenic duplication and modification of short oligopeptide-encoding DNA sequences, which might have provided a mechanism of de novo evolution of this polypeptide. PMID- 12885958 TI - Distinct changes of genomic biases in nucleotide substitution at the time of Mammalian radiation. AB - Differences in the regional substitution patterns in the human genome created patterns of large-scale variation of base composition known as genomic isochores. To gain insight into the origin of the genomic isochores, we develop a maximum likelihood approach to determine the history of substitution patterns in the human genome. This approach utilizes the vast amount of repetitive sequence deposited in the human genome over the past approximately 250 Myr. Using this approach, we estimate the frequencies of seven types of substitutions: the four transversions, two transitions, and the methyl-assisted transition of cytosine in CpG. Comparing substitutional patterns in repetitive elements of various ages, we reconstruct the history of the base-substitutional process in the different isochores for the past 250 Myr. At around 90 MYA (around the time of the mammalian radiation), we find an abrupt fourfold to eightfold increase of the cytosine transition rate in CpG pairs compared with that of the reptilian ancestor. Further analysis of nucleotide substitutions in regions with different GC content reveals concurrent changes in the substitutional patterns. Although the substitutional pattern was dependent on the regional GC content in such ways that it preserved the regional GC content before the mammalian radiation, it lost this dependence afterward. The substitutional pattern changed from an isochore preserving to an isochore-degrading one. We conclude that isochores have been established before the radiation of the eutherian mammals and have been subject to the process of homogenization since then. PMID- 12885959 TI - Comparative analysis of amino acid usage and protein length distribution between alternatively and non-alternatively spliced genes across six eukaryotic genomes. AB - Alternative splicing has been discovered in nearly all metazoan organisms as a mechanism to increase the diversity of gene products. However, the origin and evolution of alternatively spliced genes are still poorly understood. To understand the mechanisms for the evolution of alternatively spliced genes, it may be important to study the differences between alternatively and non alternatively spliced genes. The aim of this research was to compare amino acid usage and protein length distribution between alternatively and non-alternatively spliced genes across six nearly complete eukaryotic genomes, including those of human (Homo sapiens), mouse (Mus musculus), rat (Rattus norvegicus), fruit fly (Drosophila melanogaster), Caenorhabditis elegans, and bovine (Bos taurus). Our results have suggested the following: (1) across the six species, alternatively and non-alternatively spliced genes have very similar tendency for amino acids usage for not only the overall scale but also those highly expressed genes, with all of the highly expressed genes having preferred amino acids including A, E, G, K, L, P, S, V, R, T, and D. (2) For not only the overall genes but also those highly expressed ones, the average length of the protein products of alternatively spliced genes is significantly greater than that of non alternatively spliced ones. In contrast, distributions of protein lengths for the two groups of genes are very similar among all six species. Based on these results, we propose that alternatively spliced genes may have originated from non alternatively spliced ones through events such as DNA mutations or gene fusion. PMID- 12885960 TI - pANT: a method for the pairwise assessment of nonfunctionalization times of processed pseudogenes. AB - We present a method for pairwise Assessment of Nonfunctionalization Times (pANT) in processed pseudogenes. Contrary to existing methods for estimating nonfunctionalization times, pANT utilizes previously calculated probabilities of nucleotide substitution as explicit rate measurements, rather than assume that the substitution rates are the same for all nucleotides. Thus, the method allows a more accurate computation of the time that has elapsed since the nonfunctionalization of a pseudogene. Whereas existing methods require the sequence of an orthologous functional gene, which is not always at hand, pANT only uses the pairwise alignment of the gene/pseudogene pair, thus expanding the range of problems that can be tackled. To estimate evolutionary times in nonfunctional sequences, pANT measures the differences in the pairwise alignment of a gene and its paralogous processed pseudogene, using only the first and second codon positions. It assumes that, because of functional constraints, these positions in the sequence of the functional homolog have not changed since the time of nonfunctionalization of the pseudogene. Hence, the sequence of the gene may be used as the ancestor of the pseudogene. We show that the method's reliance on a detailed substitution matrix, which is derived separately for each species, makes it more accurate than existing methods. We applied pANT to the case of the unitary alpha-1,3-galactosyltransferase human pseudogene and found that our estimate of the nonfunctionalization time was in agreement with that obtained by taxonomic and paleontological considerations pertaining to the divergence between platyrrhines (New World monkeys) and cattarhines (Old World monkeys). PMID- 12885961 TI - Evidence for the contribution of LTR retrotransposons to C. elegans gene evolution. AB - LTR retrotransposons may be important contributors to host gene evolution because they contain regulatory and coding signals. In an effort to assess the possible contribution of LTR retrotransposons to C. elegans gene evolution, we searched upstream and downstream of LTR retrotransposon sequences for the presence of predicted genes. Sixty-three percent of LTR retrotransposon sequences (79/124) are located within 1 kb of a gene or within gene boundaries. Most gene retrotransposon associations were located along the chromosome arms. Our results are consistent with the hypothesis that LTR retrotransposons have contributed to the structural and/or regulatory evolution of genes in C. elegans. PMID- 12885962 TI - Nucleotide polymorphism in the RpII215 gene region of the insular species Drosophila guanche: reduced efficacy of weak selection on synonymous variation. AB - An approximately 6.9-kb region encompassing the RpII215 gene was sequenced for 24 individuals of the island endemic species Drosophila guanche. The comparative analysis of synonymous polymorphism and divergence in D. guanche and D. subobscura, two species with pronounced differences in population size, allows contrasting the nearly neutral character of synonymous mutations. In D. guanche, unlike in D. subobscura, (1) the ratio of preferred to unpreferred synonymous changes was similar for polymorphic and fixed changes, (2) the numbers of preferred and unpreferred changes, both polymorphic and fixed, could be explained by the mutational process, and (3) the estimated scaled selection coefficient for unpreferred mutations did not differ significantly from zero. Additionally, the comparative analysis revealed that both the ratio of preferred to unpreferred synonymous changes and the frequency spectrum of unpreferred polymorphic mutations differed significantly between species. All these results indicate that a large fraction of synonymous mutations in the RpII215 gene behave as effectively neutral in D. guanche, whereas they are weakly selected in D. subobscura. The reduced efficacy of selection in the insular species constitutes strong evidence of the nearly neutral character of synonymous mutations and, therefore, of the role of weak selection in maintaining codon bias. PMID- 12885963 TI - Evolutionary rate variation in anthocyanin pathway genes. AB - Over a broad taxonomic range that spans monocots and dicots, upstream enzymes of the anthocyanin pigment pathway have evolved less rapidly than downstream enzymes. In this article we show that this pattern is also evident within the genus Ipomoea. Specifically, the most upstream enzyme, chalcone synthase (CHS-D), evolves more slowly than the two most downstream enzymes, ancyocyanidin synthase (ANS) and UDP glucose flavonoid 3-oxy-glucosyltransferase (UFGT). This pattern appears not to be due to variation in mutation rates, because the CHS-D gene exhibits higher synonymous substitution rates than the genes for the other two enzymes. Codon-based tests for positive selection suggest that it has been negligible or absent in all three genes. In addition, the mean number of indel creating events is four times as high in the downstream genes as in CHS-D. Unlike the downstream genes, CHS-D also exhibits evidence of codon bias. Together, the evidence suggests that the difference in nonsynonymous substitution rates between upstream and downstream genes is due to relaxed constraint on the downstream genes rather than a greater frequency of positively selected substitutions. PMID- 12885964 TI - Nucleus-encoded, plastid-targeted glyceraldehyde-3-phosphate dehydrogenase (GAPDH) indicates a single origin for chromalveolate plastids. AB - Plastids (the photosynthetic organelles of plants and algae) originated through endosymbiosis between a cyanobacterium and a eukaryote and subsequently spread to other eukaryotes by secondary endosymbioses between two eukaryotes. Mounting evidence favors a single origin for plastids of apicomplexans, cryptophytes, dinoflagellates, haptophytes, and heterokonts (together with their nonphotosynthetic relatives, termed chromalveolates), but so far, no single molecular marker has been described that supports this common origin. One piece of evidence comes from plastid-targeted glyceraldehyde-3-phosphate dehydrogenase (GAPDH), which originated by a gene duplication of the cytosolic form. However, no plastid GAPDH has been characterized from haptophytes, leaving an important piece of the puzzle missing. We have sequenced genes encoding cytosolic, mitochondrion-targeted, and plastid-targeted GAPDH proteins from a number of haptophytes and heterokonts and found haptophyte homologs that branch within a strongly supported clade of chromalveolate plastid-targeted genes, being more closely related to an apicomplexan homolog than was expected. The evolution of plastid-targeted GAPDH supports red algal ancestry of apicomplexan plastids and raises a number of questions about the importance of plastid loss and the possibility of cryptic plastids in nonphotosynthetic lineages such as ciliates. PMID- 12885965 TI - High mutation rate of TPE repeats: a microsatellite in the putative transposase of the hobo element in Drosophila melanogaster. AB - The hobo transposable element contains a polymorphic microsatellite sequence located in its coding region, the TPE repeats. Previous surveys of natural populations of Drosophila melanogaster have detected at least seven different hobo transposons. These natural populations are geographically structured with regard to TPE polymorphism, and a scenario has been proposed for the invasion process. Natural populations have recently been completely invaded by hobo elements with three TPE repeats. New elements then appeared by mutation, triggering a new stage of invasion by other elements. Since TPE polymorphism appeared over a short period of time, we focused on estimating the mutation rate of these TPE repeats. We used transgenic lines harboring three TPE and/or five TPE hobo elements that had been evolving for at least 16 generations to search for a new TPE repeat polymorphism. We detected three mutants, with four, seven, and eight TPE repeats, respectively. The estimated mutation rate of the TPE repeats is therefore higher than that of neutral microsatellites in D. melanogaster (4.2 x 10-4 versus 6.5 x 10-6). The role of the transposition mechanism and the particular structure of the TPE repeats of the hobo element in this increase in the mutation rate are discussed. PMID- 12885966 TI - Unique mammalian tRNA-derived repetitive elements in dermopterans: the t-SINE family and its retrotransposition through multiple sources. AB - Short interspersed nuclear elements (SINEs) are dispersed repetitive DNA sequences that are major components of all mammalian genomes. They have been described in almost all lineages of Euarchontoglires (rodents, rabbits, primates, flying lemurs, and tree shrews), except in flying lemurs. Most SINE family members are composed of three distinct regions: a 5' tRNA-related region, a tRNA unrelated region, and a short tandem repeat at the 3' end that is AT-rich. The newly discovered SINE family in Cynocephalus deviates from this common structure. All 30 SINE loci analyzed in this family lack a tRNA-unrelated region and are composed exclusively of tRNA-related elements. Therefore, this novel SINE structure, described for the first time in mammalian genomes, was designated as t SINE. The t-SINE family exhibits a high copy number and is specific to flying lemurs. Three major t-SINE subfamilies could be distinguished on the basis of characteristic nucleotides, deletions, insertions, and duplications. These sequence-specific characteristics within subfamilies and sub-subfamilies reveal that they are derived copies of distinct progenitors. We present evolutionary relationships between subfamilies and compare relationships between the subfamilies and the isoleucine tRNA gene. t-SINE amplification occurred through multiple sources and is supposedly mobilized via the L1-encoded reverse transcriptase-dependent retrotranspositional mechanism in trans. PMID- 12885967 TI - Unmelanized plumage patterns in Old World leaf warblers do not correspond to sequence variation at the melanocortin-1 receptor locus (MC1R). AB - Evolutionary changes in patterns and coloration of plumage are likely to represent a major mechanism for speciation among birds, yet the molecular basis for such changes remains poorly understood. Recently much attention has focused on the melanocortin-1 receptor (MC1R) as a candidate locus for determining the level and extent of epidermal melanin deposition. We tested the hypothesis that MC1R sequence variation is associated with interspecific variation in unmelanized plumage pattern elements in Old World leaf warblers (genus Phylloscopus). This genus is characterized by a variety of plumage patterns that nonetheless vary along similar lines. Species vary in the presence or absence of pale (unmelanized) pattern elements against a dark background, and these patterns are used in species recognition and courtship. We sequenced most of the MC1R coding region for eight Phylloscopus species, representing the full range of plumage patterns found in this genus. Although MC1R sequence varied among species, this variation was not related to melanin-based plumage variation. Rather, evolution of this locus in these birds appears to be conservative. Ratios of nonsynonymous to synonymous substitutions (dN/dS) were consistently low, suggesting that strong purifying selection has operated at this locus, and likelihood ratio testing revealed no evidence of variable selective pressures among lineages or across codons. Adaptive evolution at MC1R may be constrained by the adaptive importance of plumage pattern elements in this genus. PMID- 12885968 TI - Protein evolution with dependence among codons due to tertiary structure. AB - Markovian models of protein evolution that relax the assumption of independent change among codons are considered. With this comparatively realistic framework, an evolutionary rate at a site can depend both on the state of the site and on the states of surrounding sites. By allowing a relatively general dependence structure among sites, models of evolution can reflect attributes of tertiary structure. To quantify the impact of protein structure on protein evolution, we analyze protein-coding DNA sequence pairs with an evolutionary model that incorporates effects of solvent accessibility and pairwise interactions among amino acid residues. By explicitly considering the relationship between nonsynonymous substitution rates and protein structure, this approach can lead to refined detection and characterization of positive selection. Analyses of simulated sequence pairs indicate that parameters in this evolutionary model can be well estimated. Analyses of lysozyme c and annexin V sequence pairs yield the biologically reasonable result that amino acid replacement rates are higher when the replacements lead to energetically favorable proteins than when they destabilize the proteins. Although the focus here is evolutionary dependence among codons that is associated with protein structure, the statistical approach is quite general and could be applied to diverse cases of evolutionary dependence where surrogates for sequence fitness can be measured or modeled. PMID- 12885969 TI - The cone visual pigments of an Australian marsupial, the tammar wallaby (Macropus eugenii): sequence, spectral tuning, and evolution. AB - Studies on marsupial color vision have been limited to very few species. There is evidence from behavioral, electroretinographic (ERG), and microspectrophotometric (MSP) measurements for the existence of both dichromatic and trichromatic color vision. No studies have yet investigated the molecular mechanisms of spectral tuning in the visual pigments of marsupials. Our study is the first to determine the mRNA sequence, infer the amino acid sequence, and determine, by in vitro expression, the spectra of the cone opsins of a marsupial, the tammar wallaby (Macropus eugenii). This yielded some information on mechanisms and evolution of spectral tuning of these pigments. The tammar wallaby retina contains only short wavelength sensitive (SWS) and middle-wavelength sensitive (MWS) pigment mRNAs. This predicts dichromatic color vision, which is consistent with conclusions from previous behavioral studies ( Hemmi 1999). We found that the wallaby has a SWS1 class pigment of 346 amino acids. Sequence comparison with eutherian SWS pigments predicts that this SWS1 pigment absorbs maximally (lambdamax) at 424 nm and, therefore, is a blue rather than a UV pigment. This (lambdamax) is close to that of the in vitro-expressed wallaby SWS pigment (lambdamax of 420 +/- 2 nm) and to that determined behaviorally (420 nm). The difference from the mouse UV pigment (lambdamax of 359 nm) is largely accounted for by the F86Y substitution, in agreement with in vitro results comparing a variety of other SWS pigments. This suggests that spectral tuning employing F86Y substitution most likely arose independently in the marsupials and ungulates as a result of convergent evolution. An apparently different mechanism of spectral tuning of the SWS1 pigments, involving five amino acid positions, evolved in primates. The wallaby MWS pigment has 363 amino acids. Species comparisons at positions critical to spectral tuning predict a lambdamax near 530 nm, which is close to that of the in vitro-expressed pigment (529 +/- 1 nm), but quite different from the value of 539 nm determined by microspectrophotometry. Introns interrupt the coding sequences of the wallaby, mouse, and human MWS pigment sequences at the same corresponding nucleotide positions. However, the length of introns varies widely among these species. PMID- 12885970 TI - Evolution of clonality and polyploidy in a weevil system. AB - The increased interest in asexual organisms calls for in-depth studies of asexual complexes that actively give rise to new clones. We present an extensive molecular study of the Otiorhynchus scaber (Coleoptera, Curculionidae) weevil system. Three forms have traditionally been recognized: diploid sexuals, triploid, and tetraploid parthenogens. All forms coexist in a small central area, but only the polyploid parthenogens have colonized marginal areas. Analyzing the phylogenetic relationship, based on three partial mitochondrial genes, of 95 individuals from 19 populations, we find that parthenogenesis and polyploidy have originated at least three times from different diploid lineages. We observe two major mitochondrial lineages, with over 2.5% sequence divergence between the most basal groups within them, and find that current distribution and phylogenetic relationships are weakly correlated. Quite unexpectedly, we also discover diploid clones that coexist with, and are morphologically indistinguishable from, the diploid sexual females. Our results support that these diploid clones are derived directly from the diploid sexuals. We also find that it is mainly an increase in ploidy level and not the benefits of asexual reproduction that confers to polyploid parthenogens the advantage over their diploid sexual relatives. PMID- 12885971 TI - Genic mutation rates in mammals: local similarity, chromosomal heterogeneity, and X-versus-autosome disparity. AB - The reduction of mutation rates on the mammalian X chromosome relative to autosomes is most often explained in the literature as evidence of male-driven evolution. This hypothesis attributes lowered mutation rates on the X chromosome to the fact that this chromosome spends less time in the germline of males than in the germline of females. In contrast to this majority view, two articles argued that the patterns of mutation rates across chromosomes are inconsistent with male-driven evolution. One article reported a 40% reduction in synonymous substitution rates (Ks) for X-linked genes relative to autosomes in the mouse-rat lineage. The authors argued that this reduction is too dramatic to be explained by male-driven evolution and concluded that selection has systematically reduced mutation rate on the X chromosome to a level optimal for this male-hemizygous chromosome. More recently, a second article found that chromosomal mutation rates in both the human-mouse and mouse-rat lineages were so heterogeneous that the X chromosome was not an outlier. Here again, the authors argued that this is at odds with male-driven evolution and suggested that selection has modulated chromosomal mutation rates to locally optimal levels, thus extending the argument of the first mentioned article to include autosomes. Here, we reexamine these conclusions using mouse-rat and human-mouse coding-region data. We find a more modest reduction of Ks on the X chromosome, but our results contradict the finding that the X chromosome is not distinct from autosomes. Multiple statistical tests show that Ks rates on the X chromosome differ systematically from the autosomes in both lineages. We conclude that the moderate reduction of mutation rate on the X chromosome of both lineages is consistent with male-driven evolution; however, the large variance in mutation rates across chromosomes suggests that mutation rates are affected by additional factors besides male driven evolution. Investigation of mutation rates by synteny reveals that synteny blocks, rather than entire chromosomes, might represent the unit of mutation rate variation. PMID- 12885972 TI - Responses to steroids and bronchodilators in COPD in the ISOLDE trial: the fat lady sings on. PMID- 12885973 TI - Breathing exercises and asthma. PMID- 12885974 TI - Where is SARS now? PMID- 12885975 TI - Closing the NETT on lung volume reduction surgery. PMID- 12885977 TI - Prednisolone response in patients with chronic obstructive pulmonary disease: results from the ISOLDE study. AB - BACKGROUND: A trial of corticosteroids has been recommended for all patients with chronic obstructive pulmonary disease (COPD), with the subsequent "response" determining the treatment selected. This approach assumes that patients can be reliably divided into responder and non-responder groups. We have assessed whether such a separation is statistically valid, which factors influence the change in forced expiratory volume in 1 second (FEV(1)) after prednisolone, and whether the prednisolone response predicts 3 year changes in FEV(1), health status, or number of exacerbations during placebo or fluticasone propionate treatment. METHODS: Oral prednisolone 0.6 mg/kg was given for 14 days to 524 patients with COPD before randomised treatment for 3 years with fluticasone propionate or placebo. Factors relating to change in FEV(1) after prednisolone were investigated using multiple regression. The response to prednisolone was entered into separate mixed effects models of decline in FEV(1) and health status during the 3 years of the study. RESULTS: The post-bronchodilator FEV(1) increased by a mean 60 ml (CI 46 to 74) after prednisolone with a wide unimodal distribution. Current smoking was the factor most strongly associated with the change in FEV(1) after prednisolone, with an increase of 35 ml in current smokers and 74 ml in confirmed ex-smokers (p<0.001). There was no relationship between the change in FEV(1) after prednisolone and the response to inhaled bronchodilators, baseline FEV(1), atopic status, age, or sex. The response to prednisolone, however expressed, was unrelated to the subsequent change in FEV(1) over the following 3 years on either placebo or fluticasone propionate. Regression to the mean effects explained much of the apparent prednisolone response. The significant effect of treatment on decline in health status was not predicted by the prednisolone response. CONCLUSION: Patients with COPD cannot be separated into discrete groups of corticosteroid responders and non-responders. Current smoking reduces the FEV(1) response to prednisolone. Prednisolone testing is an unreliable predictor of the benefit from inhaled fluticasone propionate in individual patients. PMID- 12885978 TI - Bronchodilator reversibility testing in chronic obstructive pulmonary disease. AB - BACKGROUND: A limited or absent bronchodilator response is used to classify chronic obstructive pulmonary disease (COPD) and can determine the treatment offered. The reliability of the recommended response criteria and their relationship to disease progression has not been established. METHODS: 660 patients meeting European Respiratory Society (ERS) diagnostic criteria for irreversible COPD were studied. Spirometric parameters were measured on three occasions before and after salbutamol and ipratropium bromide sequentially or in combination over 2 months. Responses were classified using the American Thoracic Society/GOLD (ATS) and ERS criteria. Patients were followed for 3 years with post bronchodilator FEV(1) and exacerbation history recorded 3 monthly and health status 6 monthly. RESULTS: FEV(1) increased significantly with each bronchodilator, a response that was normally distributed. Mean post bronchodilator FEV(1) was reproducible between visits (intraclass correlation 0.93). The absolute change in FEV(1) was independent of the pre-bronchodilator value but the percentage change correlated with pre-bronchodilator FEV(1) (r= 0.44; p<0.0001). Using ATS criteria, 52.1% of patients changed responder status between visits compared with 38.2% using ERS criteria. Smoking status, atopy, and withdrawing inhaled corticosteroids were unrelated to bronchodilator response, as was the rate of decline in FEV(1), decline in health status, and exacerbation rate. CONCLUSION: In moderate to severe COPD bronchodilator responsiveness is a continuous variable. Classifying patients as "responders" and "non-responders" can be misleading and does not predict disease progression. PMID- 12885979 TI - Non-volitional assessment of skeletal muscle strength in patients with chronic obstructive pulmonary disease. AB - BACKGROUND: Although quadriceps weakness is well recognised in chronic obstructive pulmonary disease (COPD), the aetiology remains unknown. In disabled patients the quadriceps is a particularly underused muscle and may not reflect skeletal muscle function as a whole. Loss of muscle function is likely to be equally distributed if the underlying pathology is a systemic abnormality. Conversely, if deconditioning and disuse are the principal aetiological factors, weakness would be most marked in the lower limb muscles. METHODS: The non volitional technique of supramaximal magnetic stimulation was used to assess twitch tensions of the adductor pollicis, quadriceps, and diaphragm muscles (TwAP, TwQ, and TwPdi) in 22 stable non-weight losing COPD patients and 18 elderly controls. RESULTS: Mean (SD) TwQ tension was reduced in the COPD patients (7.1 (2.2) kg v 10.0 (2.7) kg; 95% confidence intervals (CI) -4.4 to -1.4; p<0.001). Neither TwAP nor TwPdi (when corrected for lung volume) differed significantly between patients and controls (mean (SD) TwAP 6.52 (1.90) N for COPD patients and 6.80 (1.99) N for controls (95% CI -1.5 to 0.97, p=0.65; TwPdi 23.0 (5.6) cm H(2)O for COPD patients and 23.5 (5.2) cm H(2)O for controls (95% CI -4.5 to 3.5, p=0.81). CONCLUSIONS: The strength of the adductor pollicis muscle (and the diaphragm) is normal in patients with stable COPD whereas quadriceps strength is substantially reduced. Disuse may be the principal factor in the development of skeletal muscle weakness in COPD, but a systemic process preferentially affecting the proximal muscles cannot be excluded. PMID- 12885981 TI - Oxygen supplementation before or after submaximal exercise in patients with chronic obstructive pulmonary disease. AB - BACKGROUND: Evidence for improved exercise tolerance or relief of breathlessness by short term use of oxygen before or after exercise in patients with chronic obstructive pulmonary disease (COPD) is scant, and guidelines for this treatment are lacking despite widespread provision in the UK. METHODS: The effect of oxygenation either before or after exercise on perception of breathlessness and walk distance was studied in a group of patients with moderate to severe COPD (mean forced expiratory volume in 1 second (FEV(1)) 34% of predicted, mean 6 minute walk distance on air 283 m), all of whom desaturated by at least 4% on submaximal exercise. Oxygen (28%) or air was delivered double blind and in random order, either for 5 minutes before a standard 6 minute walk test (n=34) or for 5 minutes following the end of the test (n=18). Exercise tolerance was measured as the distance achieved and breathlessness was assessed using visual analogue scales (VAS) which were scored before and after exercise and during recovery. RESULTS: No increase in mean walk distance after oxygen (288 v 283 m) and no improvement in mean breathlessness scores (58 v 54 mm) or recovery times occurred with oxygen taken either before (177 v 184 seconds) or after exercise (182 v 151 seconds). CONCLUSIONS: This group of patients with COPD derived no physiological or symptomatic benefit from oxygen breathed for short periods before or after submaximal exercise. Domiciliary oxygen should only be prescribed for such patients if they have shown objective evidence of benefit on exercise testing. PMID- 12885982 TI - Effect of two breathing exercises (Buteyko and pranayama) in asthma: a randomised controlled trial. AB - BACKGROUND: Patients with asthma are interested in the use of breathing exercises but their role is uncertain. The effects of the Buteyko breathing technique, a device which mimics pranayama (a yoga breathing technique), and a dummy pranayama device on bronchial responsiveness and symptoms were compared over 6 months in a parallel group study. METHODS: Ninety patients with asthma taking an inhaled corticosteroid were randomised after a 2 week run in period to Eucapnic Buteyko breathing, use of a Pink City Lung Exerciser (PCLE) to mimic pranayama, or a PCLE placebo device. Subjects practised the techniques at home twice daily for 6 months followed by an optional steroid reduction phase. Primary outcome measures were symptom scores and change in the dose of methacholine provoking a 20% fall in FEV(1) (PD(20)) during the first 6 months. RESULTS: Sixty nine patients (78%) completed the study. There was no significant difference in PD(20) between the three groups at 3 or 6 months. Symptoms remained relatively stable in the PCLE and placebo groups but were reduced in the Buteyko group. Median change in symptom scores at 6 months was 0 (interquartile range -1 to 1) in the placebo group, -1 (-2 to 0.75) in the PCLE group, and -3 (-4 to 0) in the Buteyko group (p=0.003 for difference between groups). Bronchodilator use was reduced in the Buteyko group by two puffs/day at 6 months; there was no change in the other two groups (p=0.005). No difference was seen between the groups in FEV(1), exacerbations, or ability to reduce inhaled corticosteroids. CONCLUSION: The Buteyko breathing technique can improve symptoms and reduce bronchodilator use but does not appear to change bronchial responsiveness or lung function in patients with asthma. No benefit was shown for the Pink City Lung Exerciser. PMID- 12885984 TI - Resolution of bronchial inflammation is related to bacterial eradication following treatment of exacerbations of chronic bronchitis. AB - BACKGROUND: Recent studies of the role of bacteria in chronic bronchitis have shown that bacterial colonisation is associated with enhanced inflammation and that purulent acute exacerbations of chronic bronchitis (AECB) are associated with bacteria and characterised by increased inflammation. Changes in bronchial inflammation in response to the success or failure of bacterial eradication following AECB were therefore studied. METHODS: Bacterial quantitative culture and sputum markers of inflammation (myeloperoxidase (MPO), neutrophil elastase, leukotriene B4 (LTB4), sol:serum albumin ratio, and secretory leukoprotease inhibitor) were measured in patients presenting with culture positive purulent AECB and repeated 10 days and 2 months later. 41 patients provided sputum sufficient for both bacteriology and assessment of inflammation at baseline and day 10, and 46 provided sufficient sample for bacteriology, 40 of which could also be analysed for inflammation at 2 months (when clinically stable). RESULTS: At day 10, 17 of the 41 patient samples had a positive bacterial culture. In the stable state, 18 of the 46 samples had a positive culture, but with a significantly lower bacterial load than at presentation. Although there was no difference between the groups at presentation, the concentration of MPO was lower (p<0.05) in those in whom bacteria were eradicated by day 10 than in those with persisting bacteria. The LTB4 concentration was similarly lower (p<0.001) in those in whom bacteria were eradicated than in those with persistent bacteria. In the stable clinical state the concentrations of both MPO and LTB4 were lower in those in whom bacteria were eradicated than in patients with persisting bacteria. CONCLUSION: Resolution of bronchial inflammation following AECB is related to bacterial eradication. Those in whom bacteria continue to be cultured in their sputum have partial resolution of inflammation which may reflect continued stimulation by the reduced bacterial load. PMID- 12885985 TI - Short term outcome and risk factors for adverse clinical outcomes in adults with severe acute respiratory syndrome (SARS). AB - BACKGROUND: Severe acute respiratory syndrome (SARS) was diagnosed in Hong Kong in over 1700 patients between March and early June 2003. METHODS: 115 patients diagnosed with SARS were admitted to Queen Elizabeth Hospital, a large regional hospital in Hong Kong, from March 2003, of whom 100 were either discharged or were dead at 31 May. The patients were prospectively studied after admission to assess their short term outcomes and the risk factors associated with adverse outcomes, defined as death or the need for mechanical ventilation RESULTS: At the time of writing 18 patients had died, with a crude mortality rate of 15.7% and a 21 day mortality of 10% (standard error 3%). Thirty nine patients (34%) were admitted to the intensive care unit, 30 of whom (26%) required mechanical ventilation. Multivariate analysis showed that age above 60 (hazards ratio (HR) 3.5, 95% CI 1.2 to 10.2; p=0.02), presence of diabetes mellitus or heart disease (HR 9.1, 95% CI 2.8 to 29.1; p<0.001), and the presence of other comorbid conditions (HR 5.2, 95% CI 1.4 to 19.7; p=0.01) were independently associated with mortality. However, only the presence of diabetes mellitus and/or cardiac disease (HR 7.3, 95% CI 3.1 to 17.4; p<0.001) was associated with adverse outcomes as a whole. CONCLUSION: SARS is a new disease entity that carries significant morbidity and mortality. Specific clinical and laboratory parameters predicting unfavourable outcomes have been identified. PMID- 12885986 TI - Lung cancer patterns of care in south western Sydney, Australia. AB - BACKGROUND: Lung cancer is the leading cause of cancer deaths in New South Wales (NSW). There is a significantly higher incidence of lung cancer in the South Western Sydney Area Health Service (SWSAHS) than the NSW average. The aim of this study was to document patterns of lung cancer care for SWSAHS residents. METHODS: SWSAHS residents diagnosed with lung cancer in 1993 and 1996 were identified from the NSW Central Cancer Registry and their medical records reviewed. RESULTS: The study population comprised 527 patients of median age 68 years. 12% did not see a lung cancer specialist, 9% did not have a pathological diagnosis, and 28% did not receive any active treatment throughout the course of their illness. The median survival was 6.7 months and the 5 year overall survival was 8% (95% CI 6 to 10). The rates of pathological diagnosis, specialist referral, and treatment decreased with older age and poorer performance status. CONCLUSIONS: The management of lung cancer patients in SWSAHS is suboptimal. A significant proportion of patients are not receiving treatment. To improve patient care and outcomes, all lung cancer patients should be referred to a specialist for management, ideally in a multidisciplinary setting. Both consumers and general practitioners need to be educated about options available for the management of lung cancers and ageist and nihilistic attitudes need to be overcome. PMID- 12885988 TI - Impaired lung function and serum leptin in men and women with normal body weight: a population based study. AB - BACKGROUND: Impaired lung function is a risk factor for cardiovascular morbidity. Whether circulating factors are responsible for this association is unknown. A study was undertaken to determine whether leptin, a hormone that can promote atherothrombosis, is raised in individuals with impaired lung function. METHODS: Data from non-obese participants in the Third National Health, Nutrition, and Examination Survey (n=2808) were analysed to determine the relationship between circulating leptin levels and forced expiratory volume in 1 second (FEV(1)) values divided into quintiles (quintile 1, FEV(1) predicted < or =85.2%; quintile 2, 85.3-94.3%; quintile 3, 94.4-101.4%; quintile 4, 101.5-110.0%; and quintile 5, > or =110.1%). RESULTS: Serum leptin levels changed along the FEV(1) gradient. The highest leptin levels were found in quintile 1 (geometric mean (GM) 5.42; interquartile range (IQR) 3.00-9.60 fg/l) and the lowest in quintile 5 (GM 4.94; IQR 2.80-9.10 fg/l). Adjustments for age, body mass index, and other confounders strengthened this relationship. Compared with quintile 5, the odds of having an increased serum leptin level in quintiles 1, 2, 3, and 4 were 2.26 (95% confidence interval (CI) 1.54 to 3.31), 2.20 (95% CI 1.52 to 3.17), 1.46 (95% CI 1.01 to 2.09), and 1.28 (95% CI 0.90 to 1.83), respectively. CONCLUSION: Individuals with impaired lung function have raised serum leptin levels. Leptin may play a role in the pathogenesis of cardiovascular morbidity and mortality related to impaired lung function. PMID- 12885989 TI - Abnormal oesophageal motility in patients with chronic cough. AB - BACKGROUND: Although gastro-oesophageal reflux is a recognised cause of chronic cough, the role of oesophageal dysmotility is unknown. The aim of this study was to determine the prevalence of abnormal oesophageal motility in a selected group of patients with chronic cough. METHODS: Oesophageal manometry and 24 hour pH monitoring were performed in 43 patients with chronic cough, 34 of whom had symptoms suggestive of gastro-oesophageal reflux. Comparative manometric measurements were made in 21 healthy subjects. RESULTS: Nine patients with chronic cough had normal manometry and 24 hour pH. Of the remaining 34 patients, 11 (32%) had abnormal manometry alone, five (15%) had abnormal 24 hour pH monitoring alone, and in 18 (53%) both tests were abnormal. Only one patient in the control group had manometric abnormalities. CONCLUSIONS: These results point to a previously unrecognised high prevalence of abnormal oesophageal manometry in patients presenting with chronic cough. Oesophageal dysmotility may therefore be important in the pathogenesis of cough in these patients. PMID- 12885990 TI - Polymorphisms in the beta2 adrenergic receptor and bronchodilator response, bronchial hyperresponsiveness, and rate of decline in lung function in smokers. AB - BACKGROUND: Non-specific bronchial hyperresponsiveness (NSBH) is a known predictor of accelerated rate of decline in lung function in smokers. Polymorphisms of the beta(2) adrenergic receptor (ADRB2) have previously been associated with NSBH and bronchodilator response (BDR) in asthmatics. Based on these associations, we hypothesised that ADRB2 polymorphisms would be associated with NSBH and BDR as well as an accelerated rate of decline in lung function among smokers. METHODS: The prevalence of two ADRB2 polymorphisms, Arg16-->Gly and Gln27-->Glu, was examined in 587 smokers chosen from the NHLBI Lung Health Study for having the fastest (n=282) and slowest (n=305) 5 year rate of decline in forced expiratory volume in 1 second (FEV(1); mean DeltaFEV(1) -4.14 and +1.08% predicted/year, respectively). RESULTS: Contrary to our hypothesis, no ADRB2 allele or haplotype was associated with NSBH, BDR, or rate of decline in lung function. However, there was a significant negative association between heterozygosity at position 27 and a fast decline in lung function (adjusted odds ratio 0.56, 95% CI 0.40 to 0.78, p=0.0007). CONCLUSIONS: Heterozygosity at position 27 may be protective against an accelerated rate of decline in lung function. The polymorphism at position 16 does not contribute to the rate of decline in lung function, measures of NSBH, or BDR in smokers. PMID- 12885991 TI - Short term effects of airborne pollen concentrations on asthma epidemic. AB - BACKGROUND: Few studies have used time series to investigate the relationship between asthma attacks and aeroallergen levels on a daily basis. METHODS: This study, based on time series analysis adjusting for meteorological factors and air pollution variables, assessed the short term effects of different types of allergenic pollen on asthma hospital emergencies in the metropolitan area of Madrid (Spain) for the period 1995-8. RESULTS: Statistically significant associations were found for Poaceae pollen (lag of 3 days) and Plantago pollen (lag of 2 days), representing an increase in the range between the 99th and 95th percentiles of 17.1% (95% confidence interval (CI) 3.2 to 32.8) and 15.9% (95% CI 6.5 to 26.2) for Poaceae and Plantago, respectively. A positive association was also observed for Urticaceae (lag of 1 day) with an 8.4% increase (95% CI 2.8 to 14.4). CONCLUSIONS: There is an association between pollen levels and asthma related emergencies, independent of the effect of air pollutants. The marked relationship observed for Poaceae and Plantago pollens suggests their implication in the epidemic distribution of asthma during the period coinciding with their abrupt release into the environment. PMID- 12885994 TI - The pulmonary physician in critical care * Illustrative case 5: HIV associated pneumonia. PMID- 12885992 TI - Lung cancer 7: management of lung cancer in elderly patients. PMID- 12885995 TI - The pulmonary physician in critical care * Illustrative case 6: Acute chest syndrome of sickle cell anaemia. PMID- 12885996 TI - Chronic obstructive pulmonary disease * 11: fitness to fly with COPD. PMID- 12885998 TI - Intratracheal cocaine induced myocardial infarction: an unusual complication of fibreoptic bronchoscopy. AB - A 73 year old man developed chest pains 5 minutes after fibreoptic bronchoscopy. The procedure had been performed without sedation following an intratracheal injection of 5 ml 2.5% cocaine solution and xylocaine spray to the pharynx for topical anaesthesia. A 12-lead electrocardiogram showed an evolving anterior myocardial infarction. Cardiac catheterisation revealed coronary artery spasm in the proximal left anterior descending artery at the site of non-significant plaque disease. The risk factors, mechanisms, and treatment of cocaine induced myocardial infarction following intratracheal injections are discussed. PMID- 12885999 TI - Asthma guidelines. PMID- 12886000 TI - Self-management of asthma in general practice. PMID- 12886001 TI - Transoesophageal echocardiography and lung cancer staging. PMID- 12886002 TI - Atopic cough: little evidence to support a new clinical entity. PMID- 12886003 TI - Bronchoscopy in patients with suspected PCP: supine or sitting? PMID- 12886004 TI - Economics and the evolution of life histories. PMID- 12886005 TI - Short interfering RNA-mediated interference of gene expression and viral DNA accumulation in cultured plant cells. AB - Gene silencing mediated by double-stranded RNA is a sequence-specific RNA degradation mechanism highly conserved in eukaryotes that serves as an antiviral defense pathway in both plants and Drosophila. Short interfering RNAs (siRNAs), the 21- to 23-nt double-stranded intermediates of this natural defense mechanism, are becoming powerful tools for reducing gene expression and countering viral infection in a variety of mammalian cells. Here we report the use of siRNAs to target reporter gene expression and viral DNA accumulation in cultured plant cells. Transient expression of reporter genes encoding either GFP or red fluorescent protein from Discosoma was specifically reduced by 58% and 47%, respectively, at 24 h after codelivery of cognate siRNAs in BY2 protoplasts. In contrast to mammalian systems, the siRNA-induced silencing of GFP expression was transitive as indicated by the presence of siRNAs representing parts of the target RNA outside the region homologous to the triggering siRNA. Codelivery of an siRNA designed to target the mRNA encoding the replication-associated protein (AC1) of the geminivirus African cassava mosaic virus (ACMV) from Cameroon blocked AC1 mRNA accumulation by approximately 91% and inhibited accumulation of the ACMV genomic DNA by approximately 66% at 36 and 48 h after transfection. As with siRNA-induced reporter gene silencing, the siRNA targeting ACMV AC1 was specific and did not affect the replication of East African cassava mosaic Cameroon virus. This report demonstrates the occurrence of siRNA-mediated suppression of gene expression in cultured plant cells and that siRNA can interfere with and suppress accumulation of a nuclear-replicated DNA virus. PMID- 12886006 TI - Nonsteroidal antiinflammatory drugs inhibiting prostanoid efflux: as easy as ABC? PMID- 12886007 TI - Perspectives on the counterion switch-induced photoactivation of the G protein coupled receptor rhodopsin. PMID- 12886008 TI - Subcellular compartmentalization of human Nfu, an iron-sulfur cluster scaffold protein, and its ability to assemble a [4Fe-4S] cluster. AB - Iron-sulfur (Fe-S) clusters serve as cofactors in many proteins that have important redox, catalytic, and regulatory functions. In bacteria, biogenesis of Fe-S clusters is mediated by multiple gene products encoded by the isc and nif operons. In particular, genetic and biochemical studies suggest that IscU, Nfu, and IscA function as scaffold proteins for assembly and delivery of rudimentary Fe-S clusters to target proteins. Here we report the characterization of human Nfu. A combination of biochemical and spectroscopic techniques, including UV visible absorption and 57Fe Mossbauer spectroscopies, have been used to investigate the ability of purified human Nfu to assemble Fe-S clusters. The results suggest that Nfu can assemble approximately one labile [4Fe-4S] cluster per two Nfu monomers, and support the proposal that Nfu is an alternative scaffold protein for assembly of clusters that are subsequently used for maturation of targeted Fe-S proteins. Analyses of genomic DNA, transcripts, and translation products indicate that alternative splicing of a common pre-mRNA results in synthesis of two Nfu isoforms with distinct subcellular localizations. Isoform I is localized in the mitochondria, whereas isoform II is present in the cytosol and the nucleus. These results, together with previous reports of subcellular distributions of isoforms of human IscS and IscU in mitochondria, cytosol, and nucleus suggest that the Fe-S cluster assembly machineries are compartmentalized in higher eukaryotes. PMID- 12886009 TI - Crystallizing ideas about Parkinson's disease. PMID- 12886010 TI - Equality for the sexes in human evolution? Early hominid sexual dimorphism and implications for mating systems and social behavior. PMID- 12886011 TI - A decline in the levels of progesterone receptor coactivators in the pregnant uterus at term may antagonize progesterone receptor function and contribute to the initiation of parturition. AB - The molecular events that lead to the onset of labor in humans and in other mammalian species remain unclear. We propose that a decline in coactivators containing histone acetylase activity in myometrium may contribute to the onset of labor by impairing the function of the progesterone-progesterone receptor (PR) complex. As assessed by semiquantitative and real-time RT-PCR, immunohistochemistry, and immunoblotting, expression of the PR coactivators cAMP response element-binding protein (CREB)-binding protein and steroid receptor coactivators 2 and 3 was decreased in fundal uterine tissue of women in labor. Using the mouse as an animal model, we also found decreased coactivator levels in uterine tissues at term. In both human and mouse, the levels of acetylated histone H3 were also decreased in uterine tissues at term. Administration of trichostatin A, a specific and potent histone deacetylase inhibitor, to pregnant mice late in gestation increased histone acetylation and delayed the initiation of parturition by 24-48 h, suggesting the functional importance of the decline in histone acetylation in the initiation of labor. These findings suggest that the decline in PR coactivator expression and in histone acetylation in the uterus near term may impair PR function by causing a functional progesterone withdrawal. The resulting decrease in expression of PR-responsive genes should increase sensitivity of the uterus to contractile stimuli. PMID- 12886012 TI - Nitric oxide negatively regulates mammalian adult neurogenesis. AB - Neural progenitor cells are widespread throughout the adult central nervous system but only give rise to neurons in specific loci. Negative regulators of neurogenesis have therefore been postulated, but none have yet been identified as subserving a significant role in the adult brain. Here we report that nitric oxide (NO) acts as an important negative regulator of cell proliferation in the adult mammalian brain. We used two independent approaches to examine the function of NO in adult neurogenesis. In a pharmacological approach, we suppressed NO production in the rat brain by intraventricular infusion of an NO synthase inhibitor. In a genetic approach, we generated a null mutant neuronal NO synthase knockout mouse line by targeting the exon encoding active center of the enzyme. In both models, the number of new cells generated in neurogenic areas of the adult brain, the olfactory subependyma and the dentate gyrus, was strongly augmented, which indicates that division of neural stem cells in the adult brain is controlled by NO and suggests a strategy for enhancing neurogenesis in the adult central nervous system. PMID- 12886013 TI - Differential regulation of microtubule dynamics by three- and four-repeat tau: implications for the onset of neurodegenerative disease. AB - The microtubule (MT)-associated protein tau is important in neuronal development and in Alzheimer's and other neurodegenerative diseases. Genetic analyses have established a cause-and-effect relationship between tau dysfunction/misregulation and neuronal cell death and dementia in frontotemporal dementia and parkinsonism associated with chromosome 17; several mutations causing this dementia lead to increased ratios of four-repeat (4R) to three-repeat (3R) wild-type tau, and an attractive hypothesis is that the abnormally high ratio of 4R to 3R tau might lead to neuronal cell death by altering normal tau functions in adult neurons. Thus, we tested whether 3R and 4R tau might differentially modulate the dynamic instability of MTs in vitro using video microscopy. Although both isoforms promoted MT polymerization and decreased the tubulin critical subunit concentration to approximately similar extents, 4R tau stabilized MTs significantly more strongly that 3R tau. For example, 4R tau suppressed the shortening rate, whereas 3R tau had little or no detectable effect. Similarly, 3R tau had no effect on the length shortened during a shortening event, whereas 4R tau strongly reduced this parameter. Further, when MTs were diluted into buffer containing 4R tau, the MTs were stabilized and shortened slowly. In contrast, when diluted into 3R tau, the MTs were unstable and shortened rapidly. Thus, 4R tau stabilizes MTs differently and significantly more strongly than 3R tau. We suggest a "dosage effect" or haploinsufficiency model in which both tau alleles must be active and properly regulated to produce appropriate amounts of each tau isoform to maintain MT dynamics within a tolerable window of activity. PMID- 12886014 TI - Polyserase-I, a human polyprotease with the ability to generate independent serine protease domains from a single translation product. AB - We have identified and cloned a human liver cDNA encoding an unusual mosaic polyprotein, called polyserase-I (polyserine protease-I). This protein exhibits a complex domain organization including a type II transmembrane motif, a low density lipoprotein receptor A module, and three tandem serine protease domains. This unusual modular architecture is also present in the sequences predicted for mouse and rat polyserase-I. Human polyserase-I gene maps to 19p13, and its last exon overlaps with that corresponding to the 3' UTR of the gene encoding translocase of mitochondrial inner membrane 13. Northern blot analysis showed the presence of a major polyserase-I transcript of 5.4 kb in human fetal and adult tissues and in tumor cell lines. Analysis of processing mechanisms of polyserase I revealed that it is synthesized as a membrane-associated polyprotein that is further processed to generate three independent serine protease units. Two of these domains are proteolytically active against synthetic peptides commonly used for assaying serine proteases. These proteolytic activities of the polyserase-I units are blocked by serine protease inhibitors. We show an example of generation of separate serine protease domains from a single translation product in human tissues and illustrate an additional mechanism for expanding the complexity of the human degradome, the entire protease complement of human cells and tissues. PMID- 12886015 TI - A high-molecular-weight complex of membrane proteins BAP29/BAP31 is involved in the retention of membrane-bound IgD in the endoplasmic reticulum. AB - B cell antigen receptors (BCRs) are multimeric transmembrane protein complexes comprising membrane-bound immunoglobulins (mIgs) and Ig-alpha/Ig-beta heterodimers. In most cases, transport of mIgs from the endoplasmic reticulum (ER) to the cell surface requires assembly with the Ig-alpha/Ig-beta subunits. In addition to Ig-alpha/Ig-beta, mIg molecules also bind two ER-resident membrane proteins, BAP29 and BAP31, and the chaperone heavy chain binding protein (BiP). In this article, we show that neither Ig-alpha/Ig-beta nor BAP29/BAP31 nor BiP bind simultaneously to the same mIgD molecule. Blue native PAGE revealed that only a minor fraction of intracellular mIgD is associated with high-molecular weight BAP29/BAP31 complexes. BAP-binding to mIgs was found to correlate with ER retention of chimeric mIgD molecules. On high-level expression in Drosophila melanogaster S2 cells, mIgD molecules were detected on the cell surface in the absence of Ig-alpha/Ig-beta. This aberrant transport was prevented by coexpression of BAP29 and BAP31. Thus, BAP complexes contribute to ER retention of mIg complexes that are not bound to Ig-alpha/Ig-beta. Furthermore, the mechanism of ER retention of both BAP31 and mIgD is not through retrieval from a post-ER compartment, but true ER retention. In conclusion, BAP29 and BAP31 might be the long sought after retention proteins and/or chaperones that act on transmembrane regions of various proteins. PMID- 12886016 TI - Identification of Plasmodium falciparum antigens by antigenic analysis of genomic and proteomic data. AB - The recent explosion in genomic sequencing has made available a wealth of data that can now be analyzed to identify protein antigens, potential targets for vaccine development. Here we present, in the context of Plasmodium falciparum, a strategy that rapidly identifies target antigens from large and complex genomes. Sixteen antigenic proteins recognized by volunteers immunized with radiation attenuated P. falciparum sporozoites, but not by mock immunized controls, were identified. Several of these were more antigenic than previously identified and well characterized P. falciparum-derived protein antigens. The data suggest that immune responses to Plasmodium are dispersed on a relatively large number of parasite antigens. These studies have implications for our understanding of immunodominance and breadth of responses to complex pathogens. PMID- 12886017 TI - Zinc and mechanical prowess in the jaws of Nereis, a marine worm. AB - Higher animals typically rely on calcification to harden certain tissues such as bones and teeth. Some notable exceptions can be found in invertebrates: The fangs, teeth, and mandibles of diverse arthropod species have been reported to contain high levels of zinc. Considerable quantities of zinc also occur in the jaws of the marine polychaete worm Nereis sp. High copper levels in the polychaete worm Glycera dibranchiata recently were attributed to a copper-based biomineral reinforcing the jaws. In the present article, we attempt to unravel the role of zinc in Nereis limbata jaws, using a combination of position-resolved state-of-the-art techniques. It is shown that the local hardness and stiffness of the jaws correlate with the local zinc concentration, pointing toward a structural role for zinc. Zinc always is detected in tight correlation with chlorine, suggesting the presence of a zinc-chlorine compound. No crystalline inorganic phase was found, however, and results from x-ray absorption spectroscopy further exclude the presence of simple inorganic zinc-chlorine compounds in amorphous form. The correlation of local histidine levels in the protein matrix and zinc concentration leads us to hypothesize a direct coordination of zinc and chlorine to the protein. A comparison of the role of the transition metals zinc and copper in the jaws of two polychaete worm species Nereis and Glycera, respectively, is presented. PMID- 12886018 TI - Crystal structure of NF-kappaB (p50)2 complexed to a high-affinity RNA aptamer. AB - We have recently identified an RNA aptamer for the transcription factor NF-kappaB p50 homodimer [(p50)2], which exhibits little sequence resemblance to the consensus DNA target for (p50)2, but binds (p50)2 with an affinity similar to that of the optimal DNA target. We describe here the 2.45-A resolution x-ray crystal structure of the p50 RHR/RNA aptamer complex. The structure reveals that two RNA molecules bind independent of each other to the p50 N-terminal Ig-like domains. The RNA secondary structure is comprised of a stem and a stem-loop separated by an internal loop folded into a kinked helix because of the cross strand stacking of three internal loop guanines. These guanines, placed at the edge of the 3' helix, together with the major groove of the irregular 3' helix, form the binding surface for p50. Each p50 monomer uses the same surface to recognize the distorted RNA major groove as observed in the kappaB DNA/p50 RHR complex, resulting in strikingly similar interfaces. The structure reveals how the aptamer specifically selects p50 and discriminates against p65. We also discuss the physiological implications of RNA binding by (p50)2. PMID- 12886019 TI - The genome sequence of Blochmannia floridanus: comparative analysis of reduced genomes. AB - Bacterial symbioses are widespread among insects, probably being one of the key factors of their evolutionary success. We present the complete genome sequence of Blochmannia floridanus, the primary endosymbiont of carpenter ants. Although these ants feed on a complex diet, this symbiosis very likely has a nutritional basis: Blochmannia is able to supply nitrogen and sulfur compounds to the host while it takes advantage of the host metabolic machinery. Remarkably, these bacteria lack all known genes involved in replication initiation (dnaA, priA, and recA). The phylogenetic analysis of a set of conserved protein-coding genes shows that Bl. floridanus is phylogenetically related to Buchnera aphidicola and Wigglesworthia glossinidia, the other endosymbiotic bacteria whose complete genomes have been sequenced so far. Comparative analysis of the five known genomes from insect endosymbiotic bacteria reveals they share only 313 genes, a number that may be close to the minimum gene set necessary to sustain endosymbiotic life. PMID- 12886020 TI - Conjugation to gold nanoparticles enhances polyethylenimine's transfer of plasmid DNA into mammalian cells. AB - Branched polyethylenimine (PEI) chains with an average molecular mass of 2 kDa (PEI2) have been covalently attached to gold nanoparticles (GNPs), and the potency of the resulting PEI2-GNPs conjugates as vectors for the delivery of plasmid DNA into monkey kidney (COS-7) cells in the presence of serum in vitro has been systematically investigated. The transfection efficiencies vary as a function of the PEI/gold molar ratio in the conjugates, with the best one (PEI2 GNPII) being 12 times more potent than the unmodified polycation. This potency can be further doubled by adding amphiphilic N-dodecyl-PEI2 during complex formation with DNA. The resulting ternary complexes are at least 1 order of magnitude more efficient than the 25-kDa PEI, one of the premier polycationic gene-delivery vectors. Importantly, although unmodified PEI2 transfects just 4% of the cells, PEI2-GNPII transfects 25%, and the PEI2-GNPII/dodecyl-PEI2 ternary complex transfects 50% of the cells. The intracellular trafficking of the DNA complexes of these vectors, monitored by transmission electron microscopy, has detected the complexes in the nucleus <1 h after transfection. PMID- 12886021 TI - Clusterin as a biomarker in murine and human intestinal neoplasia. AB - Early detection of colorectal cancer is critical for the management of this disease. Biomarkers for early detection of several cancers have been developed and applied clinically in recent years. We have sought to discover candidate biomarkers without the restricted choice of markers placed on microarrays, and without the biological complications of genetic and environmental heterogeneity. We have compared by cDNA subtraction two genetically matched sets of mice, one developing multiple intestinal neoplasia (C57BL/6J-ApcMin) and the other tumor free (C57BL/6J). One prominent candidate biomarker, clusterin, was then subjected to a series of validation steps. In situ hybridization and immunohistochemistry were used to analyze clusterin expression at a cellular level on a series of murine intestinal and human colonic neoplasms. Elevated clusterin expression was characterized within certain regions of murine and human tumors regardless of tumor stage, location, or mode of initiation. The cells showing high clusterin levels generally lacked differentiation markers and adenomatous polyposis coli antigen. Tumor cells undergoing apoptosis expressed low levels of clusterin. Its specific expression patterns and correlation with cellular events during tumorigenesis make it a useful diagnostic tool in the mouse and a potential contributor to the set of biomarkers for early detection of human colon cancer. PMID- 12886022 TI - Skeletal muscle engraftment potential of adult mouse skin side population cells. AB - Adult bone marrow and skeletal muscle have been shown to contain a subpopulation of cells, called side population (SP) cells, that can be isolated with the fluorescence-activated cell sorter. We used a similar method to identify SP cells in the skin of adult mice. These cells express surface markers similar to SP cells isolated from skeletal muscle, but differ from bone marrow SP cells and do not express hematopoietic markers. When transplanted into nonirradiated mdx mice, nuclei from donor skin SP cells are found within myofibers that express dystrophin. Thus, adult skin SP cells can engraft in dystrophic skeletal muscle even in the absence of total body irradiation. PMID- 12886023 TI - Downregulation of superoxide dismutase activity and gene expression in cultured rat brain astrocytes after incubation with vitamin C. AB - Reactive oxygen species have been linked with neuropathological changes in the central nervous system. Epidemiological studies supported the beneficial effect of supplementation of antioxidants. Superoxide dismutase (SOD) is an endogenous enzyme which can scavenge reactive oxygen species. This study investigated the effect of supplementation with ascorbic acid (vitamin C) on the changes of SOD in cultured neurological cells. Rat brain astrocytes (RBA-1 cells) were incubated with vitamin C and divided into four groups: a control group (without vitamin C) and three treatment groups with vitamin C at 40, 80, and 160 micromol/l. After short-term (2 days) and long-term (7 days) incubation, SOD activity, SOD mRNA level by Northern blotting, and SOD protein amounts by Western blotting were measured. After 2 days of incubation, vitamin C resulted in a decrease in the activity of SOD in a concentration-dependent manner (Mn-SOD from 14.8 +/- 1.2 to 13.2 +/- 0.5 U/mg protein and Cu/Zn-SOD from 64.8 +/- 1.2 to 51.7 +/- 0.9 U/mg protein; p < 0.05), and vitamin C also attenuated the Cu/Zn-SOD mRNA level from 100 to 86.3 +/- 6.7%; p < 0.01), whereas the protein amounts of these two SODs remained unchanged. After 7 days of incubation with vitamin C, the SOD activity of RBA-1 cells decreased significantly (Mn-SOD from 14.9 +/- 0.3 to 11.8 +/- 0.3 U/mg protein and Cu/Zn SOD from 61.8 +/- 1.8 to 54.6 +/- 0.9 U/mg protein; p < 0.01), and the mRNA level was also attenuated (Mn-SOD from 100 to 86.8 +/- 8.7% and Cu/Zn-SOD from 100 to 84.7 +/- 4.8%; p < 0.01). These results suggest that 2 and 7 days of incubation with relatively high concentrations of vitamin C may downregulate activity and gene expression of SOD in cultured RBA-1 cells. PMID- 12886024 TI - Effect of endothelin on bladder contraction: potential role in bladder hyperactivity. AB - In the present study, we demonstrate that the intravenous infusion of endothelin 1 (3 and 10 ng/kg/min) causes a decrease in the mean micturition volume of rats in addition to an increase in mean arterial pressure. These effects are blocked by both the ET(A)/ET(B)-non-selective and the ET(A)-selective endothelin antagonists SB 217242 and SB 247083 respectively (both 30 mg/kg). However, it was also observed that the ET(B)-selective agonist sarafotoxin 6c (3 and 10 ng/kg/min) had similar effects on both mean arterial pressure and micturition volume. Initial experiments indicated that spontaneously hypertensive rats have a much lower mean micturition volume than normal rats. Binding studies comparing the total number and ratio of ET(A)/ET(B) receptors in spontaneously hypertensive, Wister-Kyoto and Sprague-Dawley rats revealed no significant differences in receptor expression. However, the magnitude of the response to endothelin-1 was greater in spontaneously hypertensive versus normal rats. PMID- 12886025 TI - Roles of reactive oxygen species, NF-kappaB, and peroxiredoxins in glycochenodeoxycholic acid-induced rat hepatocytes death. AB - The aim of this study was to determine the roles of reactive oxygen species (ROS), NF-kappaB and antioxidants in glycochenodeoxycholic acid (GCDC, 0-400 micromol/l, 0.5- 3 h)-induced hepatocytes death. The differential uptake of ethidium bromide and acridine orange revealed that apoptotic death occurred dose dependently in GCDC-treated hepatocytes whereas necrotic death was prominent especially at higher GCDC concentrations (> or =200 micromol/l). ROS generation measured fluorometrically either by a confocal laser microscope or by a microplate fluorescence reader was increased dose-dependently. The dose-dependent NF-kappaB activation with the significant IkappaB-alpha decrease preceded both hepatocyte cell death and the alteration of antioxidant enzymes. The Cu/Zn-SOD level among several antioxidants, we checked, remained unchanged. In contrast, the catalase level and its enzymatic activity were markedly decreased only at 400 micromol/l. The Prx I and Prx II, newly defined antioxidant enzymes reducing H(2)O(2) levels were decreased at the 200 and 400 micromol/l. These observations point to ROS generation in the GCDC-treated hepatocyte as the proximate event that triggers NF-kappaB activation, IkappaB-alpha proteolysis, Prx depletion, and finally cell death. And oxidative stress may be more related to necrotic cell death in GCDC-treated hepatocytes. PMID- 12886026 TI - Estradiol relaxes rat aorta via endothelium-dependent and -independent mechanisms. AB - The effects of estrogen on arterial function are heterogeneous with respect to vessel and/or species. We have investigated 17beta-estradiol-induced relaxation in isolated rat aorta with regard to the role of the vascular endothelium and ionic mechanisms. Estrogen induced a concentration-dependent relaxation of 46.5 +/- 7.9% and 70.1 +/- 12.2% (10(-8) and 10(-7)M), which was reduced by endothelial denudation. Furthermore, L-nitroarginine methyl ester completely abrogated this effect; however, estradiol did not relax KCl-contracted rings. Tetraethyl ammonium (1 mmol/l) completely blocked estradiol-induced relaxation. Estradiol increased [cGMP] in isolated aortic rings via NO, but did not significantly affect NOS activity in endothelial cells. Thus, estrogen can relax rat aorta in vitro via both endothelium-dependent and -independent mechanisms involving the NO/cGMP and potassium channel signaling system. PMID- 12886027 TI - Effects of endogenous histamine on seizure development of pentylenetetrazole induced kindling in rats. AB - This study was performed to investigate whether or not endogenous histamine can protect seizure development of pentylenetetrazole (PTZ)-induced kindling in rats. An intracerebroventricular (i.c.v.) injection with clobenpropit (5 and 10 microg), a representative H(3)-antagonist, significantly prolonged the onset of kindling and inhibited the seizure stages in a dose-dependent manner. Its action was significantly reversed by both immepip (2 microg, i.c.v.), an H(3)-agonist, and alpha-fluoromethylhistidine (alpha-FMH, 10 microg, i.c.v.), a selective histidine decarboxylase inhibitor. alpha-FMH (20 microg, i.c.v.) and pyrilamine (1 and 5 mg/kg i.p.), a classical H(1)-antagonist, markedly augmented the severity of seizure development of PTZ-induced kindling. Therefore, these results indicate that brain endogenous histamine plays a certain protective role on seizure development of PTZ-induced kindling in rats, and that its protective roles are mediated by H(1)-receptors. PMID- 12886028 TI - P2Y(1) receptor activation enhances the rate of rat pinealocyte-induced extracellular acidification via a calcium-dependent mechanism. AB - Pineal gland G-protein coupled P2Y(1) receptors potentiate noradrenaline-induced N'-acetylserotonin production, a long term response which occurs after 5 h incubation. In the current study we show that a short-term effect of stimulation of P2Y(1) receptors is the increase in extracellular acidification rate (ECAR), which is mediated by an increase in intracellular calcium concentration ([Ca(2+)](i)). The pD(2) values for ATP (3.06 +/- 0.12)-induced ECAR increase was significantly smaller (p < 0.01) than that for ADP (3.64 +/- 0.18), 2MeSATP (3.56 +/- 0.02) and 2MeSADP (3.65 +/- 0.13). The selective P2Y(1) receptor antagonists A3'P-5'P and A3'P-5'PS inhibited the increase in ECAR-induced by ADP. Clamping [Ca(2+)](i) with BAPTA (30 and 50 micromol/l) led to inhibition of ADP-induced increase in ECAR. Agonist and antagonist data indicate P2Y(1) activation leads to a [Ca(2+)](i)-dependent acidification of the extracellular medium. PMID- 12886029 TI - The cAMP pathway sensitizes VR1 expressed in oocytes from Xenopus laevis and in CHO cells. AB - The vanilloid receptor 1 (VR1) is a heat-activated cation channel which also responds to capsaicin and other chemical stimuli. Protein kinase C has a stimulatory effect on VR1 activity, either alone or after activation with capsaicin. The influence of the cAMP-signaling pathway on the effects of capsaicin is controversial. To clarify this, the actions of capsaicin and the modulatory effects of forskolin, pCPT-cAMP, and isobutylmethylxanthine were studied in Xenopus laevis oocytes expressing rat VR1 and in CHO cells expressing human VR1. Capsaicin activated the VR1 channel and increased the intracellular calcium concentration. The effects of capsaicin were enhanced by forskolin, pCPT cAMP, and isobutylmethylxanthine. A modulatory function of the cAMP system on VR1 activation could, therefore, modulate heat sensation and pain. PMID- 12886030 TI - Effects of camonagrel, a selective inhibitor of platelet thromboxane synthase, on the platelet-subendothelium interaction. AB - The aim of this study was to compare the effects of a new thromboxane synthase inhibitor, camonagrel, on platelet aggregation and platelet-subendothelium interaction under flow conditions, in comparison with a standard thromboxane synthase inhibitor (dazoxiben) and a cyclooxygenase inhibitor (acetylsalicylic acid). With respect to platelet aggregation in whole blood, the 50% inhibitory concentrations (IC(50)) of camonagrel were between 318 and 797 micromol/l after induction with collagen and adenosine 5'-diphosphate, respectively. For inhibition of thromboxane B(2) synthesis, the IC(50) values were 868 +/- 68 micromol/l; prostaglandin E(2) was inhibited only by acetylsalicylic acid (IC(50) for camonagrel >2,000 micromol/l), and the leukocyte 6-keto-PGF(1alpha) level was increased by camonagrel. The greatest reduction in percentage subendothelial surface occupied by platelets (mainly in the thrombi) after blood perfusion was seen after incubation with camonagrel in the range of concentrations that inhibited collagen-induced platelet aggregation. In conclusion, camonagrel reduced platelet-subendothelium interaction under flow conditions, showing this effect in a range of concentrations lower than in inhibition of platelet aggregation. PMID- 12886031 TI - Effects of RS-601, a novel leukotriene D(4)/thromboxane A(2) dual receptor antagonist, on asthmatic responses in guinea pigs. AB - The effects of 4-[4-[5,5,6,6,6-pentafluoro-1-(4-fluorobenzene sulfonamido)hexyl]phenyl]butyric acid (RS-601), a novel leukotriene D(4) (LTD(4))/thromboxane A(2) (TxA(2)) dual receptor antagonist, on bronchial asthmatic responses in guinea pigs were examined. The effects were compared with those of pranlukast (LTD(4) receptor antagonist) and S-1452 (TxA(2) receptor antagonist). RS-601 inhibited the increase in airway resistance caused by LTD(4) and TxA(2) mimetic compound, U-46619, but not by histamine. RS-601 and pranlukast but not S-1452 inhibited an antigen-induced late asthmatic response. In addition, RS-601 inhibited an antigen-induced airway hyperresponsiveness (AHR), whereas pranlukast and S-1452 had no effect on the AHR. The antigen-induced increase in inflammatory cells in airway was not affected by all examined agents. Furthermore, bacterial lipopolysaccharide-induced AHR in guinea pigs was clearly suppressed by RS-601 but not by pranlukast and S-1452. The increase in airway inflammatory cells caused by lipopolysaccharide was not affected by all three drugs. These findings indicate that RS-601 has a potent antiasthmatic efficacy, especially on AHR, but does not affect accumulation of eosinophils in the airways. PMID- 12886032 TI - Lower corneal temperature in neuroleptic-treated vs. drug-free schizophrenia patients. AB - Antipsychotic drugs (APDs) can decrease core body temperature in schizophrenia patients. Core temperature may correlate with corneal temperature and thus, we hypothesized that neuroleptic-treated schizophrenia patients would display lower corneal temperature compared with drug-free patients. Corneal temperature of 12 typical APD-treated and 9 drug-free male schizophrenia patients was assessed using a FLIR thermal imaging camera. The APD-treated patients exhibited substantially and significantly lower corneal temperature compared with the drug free patients (31.57 +/- 0.98 degrees C vs. 34.55 +/- 1.65 degrees C; p < 0.0001). Our results suggest that APDs may decrease corneal/core temperature in schizophrenia patients. The relevance of this finding to the pathophysiology of schizophrenia or to the antipsychotic effect of neuroleptics merit further investigation. PMID- 12886033 TI - Association study of serotonin transporter gene VNTR polymorphism and mood disorders, onset age and suicide attempts in a Chinese sample. AB - The human serotonin transporter (5-HTT) gene is an important candidate for the pathogenesis of mood disorders. Associations have been reported between a variable-number tandem-repeat polymorphism in intron 2 of the serotonin transporter gene (5-HTTVNTR) and mood disorders in a number of studies of Western and Chinese populations. However, no such relationships have been determined in other analogous research. To replicate these positive findings in a Chinese population and to determine the association between onset age of bipolar disorder and 5-HTTVNTR, we investigated the prevalence of this polymorphism in an independent Chinese population (83 bipolar disorder patients, 77 major depressive disorder patients and 169 controls), demonstrating no significant association between the 5-HTTVNTR polymorphism and mood disorders or age at onset. Further, no association was demonstrated between this polymorphism and suicidal history in mood disorder patients. These negative findings suggest that 5-HTTVNTR does not play a major role in the pathogenesis of mood disorder in Chinese populations. PMID- 12886034 TI - Monoamine oxidase A gene polymorphism, 5-HT 2A receptor gene polymorphism and incidence of nausea induced by fluvoxamine. AB - Selective serotonin reuptake inhibitors cause a side effect of nausea with high frequency, but there have been no accurate methods to predict its incidence. The authors first investigated whether a functional polymorphism in the monoamine oxidase A (MAOA-VNTR) and a -1438G/A polymorphism in the promoter region of the 5 HT(2A) gene were associated with the incidence of nausea induced by fluvoxamine. Fluvoxamine was administered for 6 weeks with a specific dosage plan (50-200 mg/day) in 66 Japanese major depressive patients. The frequency of MAOA-VNTR allele 1 was significantly higher in the patients without nausea than in ones with nausea in the statistical analysis including the patients whose plasma levels were below the average and who were considered to be pharmacodynamically more sensitive to nausea. This study showed that the genetic polymorphism of MAOA VNTR might affect the incidence of nausea induced by SSRIs. If this finding is replicated in other studies with more subjects, MAOA-VNTR polymorphism would be of great clinical use to predict the incidence of nausea induced by SSRIs. PMID- 12886035 TI - Decreased plasma prolactin release in euthymic lithium-treated women with bipolar disorder. AB - In order to evaluate the effect of treatment with citalopram (CIT) and lithium (Li) on hormone levels in women with bipolar disorder, morning plasma prolactin (PRL) and cortisol (CORT) were measured in 14 nonmedicated depressed patients, 13 depressed patients responding to CIT treatment, 17 euthymic patients on long-term Li treatment, and 11 healthy controls. Plasma PRL values in the Li group were significantly lower than those of the three other groups, suggesting a net inhibitory impact of augmentative effects of Li on dopaminergic activity and serotonergic neurotransmission in the central nervous system. Plasma CORT values in nonmedicated depressed patients were significantly higher than those of healthy controls, indicating hyperactivity of the hypothalamic-pituitary-adrenal system in depression, which appears to be a state-dependent phenomenon, and is normalized upon successful treatment with Li and CIT. PMID- 12886036 TI - Objective assessment of the degree of dementia by means of EEG. AB - The aim of the study was to elaborate a method to estimate the degree of cognitive impairment in Alzheimer's disease from the EEG quantitative indicators. We examined 38 unmedicated patients with a diagnosis of Alzheimer's disease, with various stages (mild, moderate, and severe) of dementia. The EEG recordings were evaluated both visually and by means of computer analysis. The EEG spectra and coherences in 6 frequency bands were calculated in 16 EEG derivations. Among various EEG indicators, a decrease in the alpha coherence and an increase in the delta coherence was found to be most significantly correlated to the degree of dementia. Combining 6 variables from the spectrum and coherence analysis by means of the multiple regression model, a high correlation (r = 0.87) between a set of EEG variables and the Mini-Mental State Examination score could be obtained. The results suggest that the EEG can supplement the clinical examination by providing an independent assessment of the degree of dementia. The results also suggest that the EEG coherences are of particular interest in dementia, being an indicator of the signal transfer between various parts of the brain cortex. PMID- 12886037 TI - Effect of riluzole on MK-801 and amphetamine-induced hyperlocomotion. AB - N-methyl-D aspartate (NMDA) antagonists, such as MK-801, and the dopamine indirect agonist amphetamine are pharmacological models used for the evaluation of putative new treatments for schizophrenia. Since the psychotomimetic effects of NMDA antagonists have recently been linked to their ability to increase glutamate release and since the glutamate release inhibitor riluzole prevented NMDA antagonist neurotoxicity, we evaluated the effect of riluzole on hyperlocomotion induced by MK-801 (0.25 mg/kg) and amphetamine (2.5 mg/kg). Mice pretreated with riluzole (3 mg/kg) did not influence baseline or MK-801-induced behavior, but 10 mg/kg produced moderate hypolocomotion alone and somewhat prolonged MK-801-induced hyperlocomotion. Pretreatment with riluzole 10 mg/kg, but not 3 mg/kg, had a moderately depressant effect both on spontaneous and amphetamine-induced locomotion. Taken together, these results suggest that riluzole would not be particularly effective as a treatment for schizophrenia and the neurotoxic and behavioral effect of NMDA antagonists do not clearly correlate. PMID- 12886038 TI - Higher plasma 5-hydroxyindoleacetic acid levels are associated with SSRI-induced nausea. AB - We investigated the association between selective serotonin reuptake inhibitors (SSRIs; paroxetine or fluvoxamine) and nausea with regard to plasma 5 hydroxyindoleacetic acid (p5-HIAA) levels. Forty-eight patients meeting the DSM IV criteria for major depressive disorder and treated with paroxetine or fluvoxamine participated in this study. p5-HIAA levels after SSRI administration were significantly higher in the nausea group than those in the nonnausea group (nausea group: 8.0 +/- 4.6 ng/ml; nonnausea group: 3.6 +/- 2.2 ng/ml; p < 0.01). On the other hand, no significant difference was found between the nausea and nonnausea group in terms of p5-HIAA levels before each drug administration. These results suggest that SSRI-induced nausea is associated with serotonergic hyperactivity in the gastrointestinal tract. PMID- 12886039 TI - The combined effects of apolipoprotein E polymorphism and low-density lipoprotein cholesterol on cognitive performance in young adults. AB - This study examined the relations of apolipoprotein E (ApoE) genotype and the low density lipoprotein (LDL) cholesterol level to cognitive performance of 57 randomly selected healthy young adults. From the ongoing population-based study of Cardiovascular Risk in Young Finns, 25 men and 32 women were ApoE genotyped and participated in mental arithmetic and reaction time tasks. In contrast to findings obtained from studies with elderly subjects, ApoE4 polymorphism was associated with better cognitive performance. In addition, LDL cholesterol moderated this association. In the ApoE4 genotype group, low LDL cholesterol was associated with good performance in the mental arithmetic test, whereas for those without ApoE4 genotype, low LDL cholesterol was associated with poor performance. Performance in the reaction time task did not differ between the ApoE groups. In conclusion, assessment of the influence of ApoE on cognitive performance may require taking additional physiological factors, such as the level of cholesterol, into account. PMID- 12886040 TI - Platelet-associated antibodies mediate protective autoimmune response in schizophrenia. AB - The peripheral blood platelets of schizophrenic patients were isolated, and the level of the platelet-associated antibodies (SPAA) was correlated with the rating scores of discrete schizophrenic symptom clusters evaluated with the Brief Psychiatric Rating Scale. Irrespective of medication and gender, symptom dependent correlations were established between the SPAA levels and the relevant psychometric scores. The results indicate a heterogeneous origin of schizophrenia and imply the involvement of an autoimmune arm as a predominantly protective immune response. PMID- 12886041 TI - The atypical antipsychotics risperidone, clozapine and olanzapine differ regarding their sedative potency in rats. AB - The atypical antipsychotics risperidone, clozapine and olanzapine were studied for their ability to antagonise apomorphine-induced stereotypy and to affect electroencephalographic (EEG) activity in rats. The compounds antagonised apomorphine-induced stereotypy with ED(50)s of 0.15 mg/kg s.c. for risperidone, 0.42 mg/kg s.c. for olanzapine and 1.3 mg/kg s.c. for clozapine. At a dose close to that required for apomorphine antagonism (0.16 mg/kg s.c.), risperidone induced only minor changes in EEG power spectral activity. At a higher dosage (0.63 mg/kg s.c.) it increased the power density in the high frequency range (9.8 18.6 Hz), characteristic of a sedative-like effect. Olanzapine and clozapine caused a dose-dependent increase in power density in all frequency bands. These effects were already present at doses (0.04 and 0.16 mg/kg s.c., respectively) 10 times below those required for apomorphine antagonism. The effects increased dose dependently and were pronounced at the doses required for apomorphine antagonism. These results in rats are in agreement with the clinical observation that risperidone has less propensity for inducing sedation at therapeutic dose levels than olanzapine and clozapine. PMID- 12886042 TI - Diabetic nephropathy: an inherited disease or just a diabetic complication? AB - Type 2 diabetic nephropathy is the most common cause of end-stage renal disease in western Europe and the United States. Although patients with overt nephropathy generally experience greater cumulative glycemic exposure, the difference in glycemic control between patients developing nephropathy and to those who did not could not be demonstrated. This observation is consistent with the finding that factors other than glycemic control are involved in the development of nephropathy. Genetic factors which specifically increase the susceptibility to nephropathy in patients with diabetes have been proposed. A range of linkage, association, and gene expression studies have been performed for revealing the genetic background of diabetic nephropathy but were not yet successful in identifying mutations which could explain the development of diabetic nephropathy in the majority of diabetic patients. Because of relatively small case numbers of all studies being performed so far, conclusions from those studies are limited. With the development of better technologies for an affordable genomewide association study using thousands of markers, it might become possible to unravel the genetic susceptibility factors for diabetic nephropathy. Comparing the expression levels of thousands of genes in patients and controls may identify key players in the pathogenesis of diabetic nephropathy and targets for pharmacologic intervention in the future. PMID- 12886043 TI - Epidermal growth factor regulates Ca2+ uptake in primary cultured renal proximal tubule cells: involvement of cAMP, PKC and cPLA2. AB - Epidermal growth factor (EGF) is known to play an important role in modulating renal transport functions. Thus, we investigated the effect of EGF on Ca(2+) uptake and its related signals in the primary cultured rabbit renal proximal tubule cells. EGF (50 ng/ml, 1 h) stimulated Ca(2+) uptake. Its effect was blocked by AG 1478 (an EGF receptor antagonist), genistein or herbimycin A (tyrosine kinase inhibitors). EGF increased intracellular cAMP level and SQ 22536 (an adenylate cyclase inhibitor), Rp-cAMP (a cAMP analogue), or PKI (a protein kinase A inhibitor) blocked the EGF-induced stimulation of Ca(2+) uptake. EGF induced stimulation of Ca(2+) uptake was also blocked by neomycin or U-73122 (phospholipase C inhibitors), staurosporine, H-7, or bisindolylmaleimide I (protein kinase C inhibitors), nifedipine or methoxyverapamil (L-type Ca(2+) channel blockers). It increased IPs formation by 167 +/- 5% compare to control within 90 s. On the other hand, EGF increased [(3)H]-arachidonic acid release, which was significantly blocked by PKC inhibitors. In addition, PGE(2), one of cyclooxygenase metabolites, and 5,6-EET, one of cytochrome P-450 metabolites, increased Ca(2+) uptake. These results suggest that cAMP, PLC/PKC, and PLA(2) are involved in EGF-induced stimulation of Ca(2+) uptake. PMID- 12886044 TI - Role of Nitric oxide in the renal and systemic vasodilatory responses to platelet activating factor in the rat, in vivo. AB - BACKGROUND/AIM: Chemical mediator(s) involved in the renal vasodilatory and systemic hypotensive effects of platelet-activating factor (PAF) remain unresolved. Because nitric oxide (NO) and PAF have many similar cardiovascular actions, we examined whether endogenous NO contributes to the renal and systemic actions of PAF. METHODS: PAF was administered into the renal arterial or systemic venous circulation of anesthetized rats. The change in renal blood flow (RBF) and/or mean arterial blood pressure (MAP) was recorded in the absence and presence of NO synthase inhibition. RESULTS: Transient exposure of the renal vacsular bed to intrarenal PAF boluses (1-10 ng kg(-1)) resulted in an immediate increase in RBF that was partially inhibited by the intrarenal administration of a NO synthase inhibitor, whereas the ensuing rapid fall in MAP was unaffected by NO synthase blockade. A sustained exposure to intrarenal PAF infusion (2.5 ng min(-1) kg(-1)) in intrarenal NO synthase inhibitor-treated rats (hypertensive with vasoconstricted kidneys) had no effect on RBF and MAP, which was in contrast to the increase in RBF and modest fall in MAP in control intrarenal vasopressin treated rats (hypertensive with vasoconstricted kidneys). To dissociate the influence of systemic hypotension on the RBF response, rat kidneys were denervated and MAP elevated by a pressor agent whilst maintaining renal arterial blood pressure constant. Subsequent intrarenal PAF infusion at 2.5 and 10 ng min( 1) kg(-1) resulted in a substantial rise in RBF and modest fall in MAP, responses that were abolished by intrarenal NO synthase inhibition at the lower intrarenal PAF infusion or abolished/attenuated at the higher intrarenal PAF infusion. Additional experiments administered drugs intravenously to exclude the possibility that the modified MAP response by NO synthase inhibition was related to the drugs being administered into the kidney. Intravenous PAF boluses (0.1-1 microg kg(-1)) resulted in a transient fall in MAP that was independent of the NO pathway, whereas the systemic hypotension induced by intravenous PAF infusion (10 ng min(-1) kg(-1)) was greatly attenuated by NO synthase inhibition. CONCLUSION: NO has a major role in mediating the renal and peripheral vasodilatory responses induced by a sustained exposure to PAF in the rat, whereas NO's overall contribution was less or absent during transient PAF exposure. PMID- 12886045 TI - Four additional CLCN5 exons encode a widely expressed novel long CLC-5 isoform but fail to explain Dent's phenotype in patients without mutations in the short variant. AB - BACKGROUND: Dent's disease is caused by mutations in the CLCN5 gene coding for the chloride channel CLC-5. However, sequencing of CLCN5 exonic regions in some patients presenting with low-molecular-weight proteinuria and hypercalciuria - the hallmarks of Dent's disease - failed to identify causative mutations. AIM: Given the observation that some species harbour a CLCN5 mRNA encoding an extended CLC-5 aminoterminus compared with the so far known human form, we worked on the presumption that an orthologous (longer) CLCN5 transcript is also present in humans and that our patients may have mutations herein. METHODS: Extensive databank mining, reverse transcription polymerase chain reaction (RT-PCR) and automated sequencing were used in the search for novel CLCN5 transcripts. The human CLCN5 gene was investigated in 7 patients out of five families by direct automated sequencing of PCR-amplified DNA products. RESULTS: Two new human CLCN5 transcripts expressed in kidney and various other tissues could be identified. These arise from a novel site of transcription initiation, alternative splicing and the use of four additional CLCN5 exons. If being translated, both these mRNAs would lead to an enlarged CLC-5 protein consisting of 816 amino acids by adding 70 aminoterminal residues to the so far known 746-amino-acid-long isoform. Sequence analysis of the henceforward 17 CLCN5 exons revealed no mutation in the patients with a phenotype resembling Dent's disease. CONCLUSIONS: Despite the identification of further targets to explain Dent's disease, the molecular defect in our patients remains to be elucidated. Hence, their phenotype may be explained by mutations that affect so far unknown regulating elements of the CLCN5 gene or another gene(s), probably encoding CLC-5 accessory protein(s). PMID- 12886046 TI - Effects of LDL apheresis and vitamin E-modified membrane on carotid atherosclerosis in hemodialyzed patients with arteriosclerosis obliterans. AB - BACKGROUND: Hemodialysis patients manifest accelerated atherosclerosis. Hemodialysis is associated with oxidative stress, which can be partially prevented with the use of a vitamin E-coated dialyzer. Adsorption of low-density lipoprotein (LDL) has been applied in the treatment of arteriosclerosis obliterans (ASO). The aim of the present study was to determine whether the vitamin E-coated dialyzer and/or LDL apheresis affects carotid atherosclerosis in hemodialysis patients with ASO. METHODS: Thirty hemodialysis patients with ASO were divided into four treatment groups: treatment with conventional cellulose or synthetic membranes (group A, n = 12), treatment with vitamin E-coated membrane (group B, n = 7), treatment with conventional membrane and LDL apheresis (group C, n = 6), and treatment with vitamin E-coated membrane and LDL apheresis (group D, n = 5). Carotid artery intima-media thickness (IMT) and arterial stiffness assessed by pulse wave velocity (PWV), plasma C-reactive protein (CRP) and interleukin (IL)-6 were measured before and 10 weeks after treatment and compared between groups. All values were referred to measurements after LDL apheresis. RESULTS: IMT and PWV, plasma CRP and IL-6 showed little change in group A throughout the experimental period. These decreased slightly from the baseline value in group B, but the change was not significant. In group C, IMT decreased from 1.12 +/- 0.24 to 1.02 +/- 0.18 mm (p < 0.05), and PWV decreased from 2,266 +/- 380 to 1,968 +/- 342 cm/s (p < 0.05). Plasma CRP and IL-6 concentrations also decreased significantly compared with baseline (p < 0.05). In group D, IMT decreased from 1.18 +/- 0.26 to 0.92 +/- 0.18 mm (p < 0.01), and PWV decreased from 2,284 +/- 390 to 1,786 +/- 284 cm/s (p < 0.01). Plasma CRP and IL-6 levels also decreased significantly compared with baseline (p < 0.01). CONCLUSION: These data suggest that LDL apheresis and the vitamin E-coated membrane dialysis in combination may prevent further progression of atherosclerosis in hemodialysis patients with ASO. PMID- 12886047 TI - Tolerability and efficacy of multidose epoetin beta (Reco-Pen) for subcutaneous administration in patients with anemia due to renal failure. AB - AIMS: To assess the tolerability, safety and efficacy of the epoetin beta multidose cartridge formulation, self-administered subcutaneously via a pen device (Reco-Pen), in adult patients with renal anemia. METHODS: Patients receiving maintenance epoetin therapy were switched to the subcutaneous (SC) multidose formulation of epoetin beta (NeoRecormon). The frequency of adverse events, local tolerability, and changes in blood pressure and laboratory variables were recorded. Hematologic parameters, transfusion requirements and epoetin beta dosage were also assessed. RESULTS: A total of 406 patients were entered in the intention-to-treat analysis. Mean treatment duration was 82.3 days. Fifty patients (12.3%) withdrew from the study; 14 (3.4%) discontinued because of adverse events. Treatment was well tolerated, with adverse events considered probably related to treatment in only 5 cases, and 1 case of local intolerability. There were no clinically significant changes in blood pressure or laboratory variables, and no changes in hematologic parameters or transfusion requirements. Unexpectedly, the epoetin beta dose was reduced by almost one-third in patients previously maintained on SC epoetin. CONCLUSION: SC administration of this multidose epoetin beta formulation with the Reco-Pen device was well tolerated and effective. It is possible that the improved capacity to individualize dose may have contributed to the considerable reduction in SC epoetin beta dosage requirement. PMID- 12886049 TI - Efficacy of 8-bromoguanosine against murine cutaneous leishmaniasis induced with Leishmania amazonensis. AB - BACKGROUND AND METHODS: In this study, we investigated the effect of 8 bromoguanosine (8Brguo), an immunostimulatory compound, in a murine model of experimental cutaneous leishmaniasis. RESULTS: The results indicated that treatment by intraperitoneal administration of 8Brguo (150 mg/kg once daily for 1 week and thereafter once weekly for 1 month) diluted in Tween-80 produces an effect similar to that of treatment with the classical antimonial drug, Glucantime, in Leishmania amazonensis-infected CBA/J mice. While complete cure did not occur, animals treated with the nucleoside had small lesions in the infected footpad and delayed surface ulceration. Studies involving examination of serum glutamic pyruvic transaminase activity, concentration of urea, blood pathology and histology of the spleen, kidney and liver showed no apparent toxicity of the nucleoside in treated mice. CONCLUSION: The protective in vivo effect during murine leishmaniasis as well as the lack of apparent toxicity of 8Brguo as attested by blood/serum pathology and histology from treated mice encourage further studies of C8-substituted guanine ribonucleosides, such as 8Brguo, as new leishmanial drugs and as modifiers of the immunological response to combat infections with intracellular pathogens. PMID- 12886050 TI - Effect of linezolid on the phagocytic functions of human polymorphonuclear leukocytes. AB - The effect of linezolid on the phagocytic and bactericidal functions of human polymorphonuclear neutrophils (PMN) against gram-positive cocci was evaluated. Preincubation (30 min; 37 degrees C) of PMN with different concentrations of linezolid (2, 10 and 20 mg/l) did not significantly affect the phagocytosis of either Staphylococcus aureus (methicillin susceptible and resistant) or Enterococcus faecalis (vancomycin susceptible and resistant). Overnight exposure of vancomycin-resistant E. faecalis to 1/4 minimum inhibitory concentrations (MIC) of linezolid slightly increased phagocytosis by PMN. Preincubation of the other three strains with 1/4 MIC of linezolid did not affect phagocytosis by these cells. Preincubation of PMN (30 min; 37 degrees C) using different extracellular concentrations of linezolid (2, 10 and 20 mg/l) did not affect their production of either superoxide or hydrogen peroxide radicals. In conclusion, linezolid did not affect the phagocytic and bactericidal functions of human PMN. PMID- 12886051 TI - The penetration of moxifloxacin into the pancreas of male rats in experimental acute necrotizing pancreatitis. AB - Infectious complications of acute necrotizing pancreatitis (ANP) determine the extent of multiorgan failure and account for 80% of deaths. Prophylactic use of antibiotics can reduce the incidence of these complications. However, the actual indication as well as choice of drug remains a controversial matter. We examined the penetration of moxifloxacin, a new broad-spectrum fluoroquinolone, in healthy and inflamed pancreatic tissue in rats after inducing ANP. The concentration of moxifloxacin in pancreatic tissue and serum was determined 10, 30, 60 and 240 min after the administration of moxifloxacin (5 mg/kg, i.v.). Mean serum concentrations 10 min after administration in rats with ANP were 1,886 ng/ml versus 1,805 ng/ml in healthy controls, and these values decreased to 350 versus 222 ng/ml, respectively, after 240 min. Corresponding concentrations in pancreatic tissue were in the mean 2-3 times higher. PMID- 12886052 TI - Target affinities of faropenem to and its impact on the morphology of gram positive and gram-negative bacteria. AB - Faropenem is a new oral beta-lactam antibiotic unique from carbapenems and other available beta-lactams. Determinants of the in vitro activity of beta-lactam antibiotics include affinity to penicillin-binding proteins (PBPs) and beta lactamase stability. In this study, the binding affinity of faropenem to various PBPs and its impact on the morphology of Staphylococcus aureus and Escherichia coli were evaluated. In general, faropenem demonstrated high binding affinity to high-molecular-weight PBPs but low affinity to low-molecular-weight PBPs. In S. aureus and Streptococcus pneumoniae, faropenem exhibited high binding affinity to PBP1, followed by PBP3 and PBP2. In E. coli, faropenem showed the highest affinity for PBP2, followed by PBP1A, PBP1B, PBP3 and PBP4. In Proteus vulgaris, binding was highest to PBP4, followed by PBP1A, PBP2 and PBP3. In Serratia marcescens, faropenem bound preferentially to PBP2 and PBP4. Exposure of S. aureus to faropenem at minimum inhibitory concentrations (MICs) of 1/8 or 1/4 resulted in irregular septum formation. At 1x MIC or higher, a larger number of lysed cells were observed. Exposure of E. coli to 1/8x MIC or 1/4x MIC also induced changes in cellular shape; the normal rod-shaped form changed to a spherical form in a time-dependent manner. After exposure of E. coli to 1x MIC for 2 h, bulging-shaped E. coli cells were observed and after 4 h of exposure cell lysis was demonstrated. In the presence of 4x MIC, spheroplast-like forms and cell lysis were observed. The morphological changes triggered by faropenem are in agreement with the PBP binding affinities reported. Thus, the high binding affinities of faropenem to PBPs from gram-negative and gram-positive bacteria are mirrored by its pronounced and concentration-dependent bactericidal effect. PMID- 12886053 TI - Comparative in vitro activity of three fluoroquinolones against clinical isolates by E test. AB - The in vitro susceptibility of levofloxacin, ciprofloxacin and moxifloxacin against several gram-positive and gram-negative clinical isolates was tested by E test. We found that the MIC(50) and MIC(90) values against all members of the Enterobacteriaceae family except Serratia were <0.5 mg/l for ciprofloxacin and levofloxacin (MIC range 0.006-32 mg/l) based on the in vitro susceptibility data. The susceptibility rates for ciprofloxacin and levofloxacin were more than 85% for Escherichia coli, citrobacter, enterobacter cloacae, enterobacter aerogenes and Klebsiella pneumoniae, although Serratia and Acinetobacter exhibited more or less similar susceptibility rates (about 80%). Pseudomonas aeruginosa demonstrated significant resistance to fluoroquinolones (MIC(90) >32 mg/l) and decreased bactericidal rates (<65%) to levofloxacin and ciprofloxacin. Respiratory pathogens such as Streptococcus pneumoniae and Haemophilus influenzae were highly susceptible (100%) to levofloxacin and moxifloxacin. The ineffectiveness of fluoroquinolones for treating coagulase-positive Staphylococcus aureus was demonstrated by poor in vitro susceptibility rates with levofloxacin (52%) and moxifloxacin (57%). Coagulase-negative staphylococci demonstrated significantly decreased bactericidal rates to levofloxacin (21%), while the in vitro susceptibility to moxifloxacin was higher (66%) than that to levofloxacin. We propose that the beneficial effect of inclusion of any of these three fluoroquinolones in treating Enterococcus infections is marginal, as demonstrated by significantly reduced susceptibility rates (<32%). These data demonstrate the utility of fluoroquinoles to treat several gram-negative bacterial infections (with the exception of Acinetobacter and P. aeruginosa), as well as S. pneumoniae and H. influenzae. PMID- 12886054 TI - Antimicrobials as potential adjunctive agents in the treatment of biofilm infection with Staphylococcus epidermidis. AB - BACKGROUND: This study was performed to evaluate the interaction of erythromycin or rifampin with vancomycin against Staphylococcus epidermidis biofilms. METHODS: Biofilm-forming S. epidermidis strains (ATCC 35983, 35984) and polyurethane (PU) sheets were incubated for the formation of bacterial biofilms. Biofilms on PU sheets were treated with various antibiotic regimens. The number of viable bacteria on the sheets was counted. RESULTS: While erythromycin or vancomycin alone did not significantly reduce the biofilm bacterial concentration, the combination of vancomycin and erythromycin resulted in a clear reduction in bacterial concentration compared with the single agents. Rifampin was the most active single agent against biofilm-forming S. epidermidis, while the combination of rifampin and vancomycin showed further reduction in bacterial concentration. CONCLUSIONS: These data suggest that the combination of erythromycin or rifampin and vancomycin is more effective than vancomycin alone in the treatment of biofilm infections caused by biofilm-forming S. epidermidis. PMID- 12886055 TI - Oxaliplatin with weekly bolus 5-fluorouracil and leucovorin in pretreated advanced colorectal cancer patients: a phase II study. AB - BACKGROUND: The purpose of this study was to determine the efficacy and toxicity of oxaliplatin in combination with weekly bolus 5-fluorouracil (5-FU) and leucovorin (LV) in patients with 5-FU-pretreated advanced colorectal cancer. METHODS: A total of 39 patients with documented 5-FU-resistant advanced colorectal cancer were enrolled. All 39 patients had previously received weekly high-dose 5-FU/LV (2,600 mg/m(2) 5-FU plus 100 mg/m(2) LV as 24-hour infusion) as first-line chemotherapy for metastatic disease. Oxaliplatin (85 mg/m(2)) was delivered as an intravenous infusion over 2 h on days 1 and 15, while 5-FU (500 mg/m(2)) and LV (20 mg/m(2)) were administered as an intravenous bolus on days 1, 8 and 15. One treatment course consisted of 3 consecutive weeks of therapy followed by a 1-week rest. RESULTS: In an intent-to-treat analysis, the objective response rate for the 39 patients was 20.5% (95% confidence interval, 7.2-33.8%). The disease was stable in 19 patients (48.7%), and progressive in 11 (28.2%). The median survival for all 39 patients was 8.9 months. The median time to progression was 5.0 months. Grade 3/4 neutropenia occurred in only 1 patient (2.6%), and grade 2 and 3/4 peripheral neuropathy occurred in 10 (25.6%) and 5 (12.8%) patients, respectively. CONCLUSION: Oxaliplatin in combination with weekly bolus 5-FU/LV is also active in patients with advanced colorectal cancer where first-line treatment has failed. PMID- 12886056 TI - Combination chemotherapy with irinotecan and ifosfamide as second-line treatment of refractory or sensitive relapsed small cell lung cancer: a phase II study. AB - This phase II study was conducted to investigate the efficacy and safety of irinotecan (CPT-11) and ifosfamide as second-line chemotherapy for relapsed small cell lung cancer (SCLC). Eligibility criteria included histologically or cytologically confirmed SCLC, prior chemotherapy including platinum + etoposide, and measurable or evaluable disease. CPT-11 (80 mg/m(2)) was administered intravenously on days 1, 8 and 15, while ifosfamide (1.5 g/m(2)) was given on days 1 through 3 every 4 weeks. Thirty-four patients (29 men) with a median age of 69 years (range 42-77) and a median Eastern Cooperative Oncology Group (ECOG) performance status of 1 (range 0-2) were enrolled. The response rate was 52.9% (95% confidence interval: 29.8-64.9%) with 2 complete responses and 16 partial responses. Our analyses of prognostic factors showed risk factors assessed before receiving second-line chemotherapy, which were the number of metastatic sites, performance status and the type of relapse. WHO grade 3-4 neutropenia was recorded in 52.9% of the patients, grade 3 diarrhea in 5.9%. The combination of CPT-11 and ifosfamide demonstrated clinical efficacy in relapsed SCLC with a favorable toxicity profile, particularly for performance status 0-1 and sensitive cases with only one metastatic site. PMID- 12886057 TI - Ampicillin, gentamicin and teicoplanin as antimicrobial therapy for recurrent Streptococcus agalactiae and Enterococcus faecalis endocarditis in an intravenous drug abuser with HIV infection. AB - Infective endocarditis associated with human immunodeficiency virus (HIV) infection occurs almost exclusively in intravenous (i.v.) drug users and usually involves the tricuspid valve, with an increased mortality rate among patients with a severe degree of immunosuppression. The first reported case of recurrent tricuspid endocarditis sustained by Streptococcus agalactiae and Enterococcus faecalis in an i.v. drug addict during HIV infection is presented. Antimicrobial therapy with i.v. ampicillin, gentamicin and teicoplanin led to complete clinical and echocardiographical recovery. PMID- 12886059 TI - Levoflaxin and Streptococcus pneumoniae. PMID- 12886058 TI - Lung, pleural and colon actinomycosis in an immunocompromised patient: a rare form of presentation. AB - Actinomycosis is caused by gram-positive filamentous organisms of the genus Actinomyces, which may spread through trauma. Most commonly, it is a cervicofacial disease due to dental infection or a thoracic disease secondary to aspiration of foreign bodies. Primary abdominal infection usually follows some form of mucosal disruption. Any organ of the human body may be involved so that a wide range of symptoms may be present. We report a rare form of actinomycosis involving the lung, pleura and colon concomitantly in an immunocompromised patient. A fine needle aspiration from a lung lesion detected the characteristic sulfur granules, and a pleural effusion culture confirmed the diagnosis. Clinical manifestations and treatment are discussed. Actinomycetes are rarely opportunistic agents in immunocompromised patients; thus the disease deserves special attention in those patients. PMID- 12886060 TI - [Effect of aflatoxin B1 on ultrastructural changes of biliary epithelial cells in mice experimentally infected with Clonorchis sinensis] AB - The present study was carried out to examine the effect of a carcinogen, aflatoxin B1 on the ultrastructural changes of biliary epithelial cells in mice infected with Clonorchis sinensis. A total of 93 male albino mice(BALB/c strain) was divided into 3 groups; group I, treated with 1.0 ppm aflatoxin B1 for 12 weeks; group II, given 50 C. sinensis metacercariae, and group III, given 50 metacercariae and treated with 1.0 ppm aflatoxin B1 for 12 weeks. Three mice served for untreated-uninfected controls. From 4 weeks after the treatment and/or infection, three mice from each group were sacrificed at 4 week intervals up to the 40th week, and their hepatobiliary tissues were prepared for transmission electron microscopy. The most prominent ultrastructural changes in group I were remarkable enlargement of nuclear size, separation of nucleolus, dispersed chromatin granules in nuclei and increased dense granules along the inner membrane of nuclei. In the cytoplasm there was slight proliferation of mitochondria and endoplasmic reticulum (ER) at earlier stage. At the 12th week separation of fibrillar and granular components of the nucleolus was a characteristic finding. As the time elapsed, epithelial cells showed flattened cuboidal form and a tendency of atrophy. Most of the nuclei were elongated and polygonal in shape. In group II the appearance of elaborate interwoven folds of lateral cytoplasm forming a labyrinth of interconnected intercellular space and variety in nuclear shape were the prominent findings at earlier stage. The cytoplasm showed slight proliferation and dilatation of mitochondria and ER, and a small number of mucin droplets. In the basement membrane scanty fibrous cells were seen. With time, variety in nuclear shape, marked proliferation and dilatation of rough ER and some collagen fibrils were demonstrated. Other features of intracellular organelles and mucin droplets persisted. In group III cuboidal epithelial cells showed their remarkably enlarged and irregular nuclei, increased chromatin granules in the nuclei, separated nucleoli, proliferated and dilated rough ER. With time, sequestered mitochondria showed bleb-like evaginations which lacked cristae and dense matrix, and were limited by a single membrane. Since the 20th week, microvilli were relatively scanty and poorly developed. Organelles and inclusions in the cytoplasm of metaplastic cells were poor. Nuclei were variable in shape. The most prominent changes at later stage were separation of nuclei from the cytoplasm, and appearance of numerous and irregularly angled electron dense granules in the nuclei. PMID- 12886061 TI - Histopathological changes of the liver after praziquantel treatment in Clonorchis sinensis infected rabbits. AB - The rabbits were infected with Clnorchis sinensis and were treated with praziquantel at the dose of 50 mg/kg x 2 x 2 days afer 1, 2, 4, 8 weeks and 7 months from the infection. Their livers were observed histopathologically 1, 4 and 12 weeks after treatment. The findings are summarized as below: The changes of the liver in control rabbits were relatively mild until 2 weeks after infection. However, widening and thickening of bile ducts, proliferation of biliary epithelium and periductal fibrosis were moderate after 4 weeks from infection and those changes were severe after 8 weeks and 7months. Goblet cell metaplasia was found after 8 weeks from infection. The mild changes of 2-week infection group were completely recovered by 4 weeks after the treatment. In the groups of 4 or more weeks after infection, the changes of bile ducts became milder in the degree after the treatment, but were still found 12 weeks after the treatment. As the infection duration was passed, more severe changes were observed after the treatment. In this context, it is concluded that the liver changes of acute clonorchiasis in the early two weeks are reversible by treatment while chronic biliary epithelial changes are irreversible. Therefore, early treatment should be recommended as possible to minimize the remaining histopathological changes of liver in clonorchiasis. PMID- 12886062 TI - [Larvicidal action of liquid nitrogen against metacercariae of Clonorchis sinensis] AB - The author investigated the larvicidal action of liquid nitrogen against the metacercariae of Clonorchis sinensis, with an observation on the freshness and change of taste in the flesh of fishes. The results obtained were as follows: The metacercariae in the flesh of Pseudorasbora parva, which were treated with liquid nitrogen (-192C), were not killed within 10 seconds, but completely killed over 30 seconds. In comparison, the metacercariae in the flesh of P. parva kept in a refrigerator (-12C) were killed only in 84 percent in 10-hour exposure group. The freezing speed of fishes by liquid nitrogen was 4 min. (') and 15 seconds(") for Cyprinus carpio, 1'22" for Carassius carassius and only 30", for Pseudorasbora parva. As for the freshness and taste of raw fresh water fishes, they were not deteriorated after the treatment with liquid nitrogen. In animal infection experiment of C. sinensis metacercariae after freezing, they were found not infective after they were treated with liquid nitrogen. From the results, it is inferred that the treatment of fresh water fishes for longer than 30 seconds with liquid nitrogen is helpful to reduce the possibility of C. sinensis infection without alteration of the freshness and taste of fishes. PMID- 12886063 TI - [Ultrastructural studies on the surface of Paragonimus westermani metacercaria] AB - The present study was undertaken to demonstrate the surface structure of Paragonimus westermani metacercaria in Korea with special reference to the distribution of sensory papillae. Metacercariae were isolated from crayfish, one of the second intermediate host of P. westermani in Bogil island, Chollanam-do (Province), Korea, where has been known as an endemic area of human paragonimiasis. Isolated metacercariae were excysted and examined with light, scanning and transmission electron microscopes for morphological features. On the surface of metacercariae, three types of sensory papillae were identified. Large domed papillae (3-5 micrometer), which were covered with wrinkled plasma membrane of the worm, were distributed on the oral and ventral suckers only. On the oral sucker, these large domed papillae were 12-13 in number. On the other hand large domed papillae on the ventral sucker were constantly 6 in number and hexagonal in distribution. Small domed papillae (2-3 micrometer), of which surface was more smooth than those of large ones, were distributed symmetrically on the ventral (30-32 pairs) and dorsal surfaces (40-42 pairs). Ciliated papillae (0.8-1.5 micrometer) were observed about 5-6 in number around the oral sucker and 3-5 pairs each on the ventral and dorsal surface of the body. Single pointed spines covered the entire surface of the body except around the excretory pore. Spines on the anterior part of the body were 0.9-2.0 micrometer in length and 45-55/100 square micrometer in number, and were gradually reduced in length (0.4-1.4 micrometer) and in number (12-27/100 square micrometer) toward the posterior part. The body wall of P. westermani metacercariae was consisted with anucleated syncytium layer, fibrous interstitial layer and musclar layer. In the anucleated syncytium, biconcave (0.15-0.55 micrometer) and spherical (0.08-0.16 micrometer) secretory granules, which were transferred from epidermal cells via protoplasmic tubules, mitochondria and ribosomes, were observed. Spines originated around the basement membrane protruded externally. Epidermal cells were consisted with a nucleus and a cytoplasm, and connected to syncytium with protoplasmic tubules. In the cytoplasm many secretory granules, mitochondria, Golgi complex, endoplasmic reticula, ribosomes and lipid droplets were observed. PMID- 12886064 TI - Detection of circulating antigens in rats experimentally infected with Paragonimus westermani by ELISA. AB - Circulating antigens in rats experimentally infected with Paragonimus westermani were examined by ELISA. From a total of 22 albino rats, each fed with 25 metacercariae, blood samples were collected until 12 weeks after infection. The specific antibodies against P. westermani in the serum of an infected cat were purified by ammonium sulfate precipitation, DEAE anion-exchange chromatography and affinity chromatography serially. So-called double antibody sandwich ELISA method was used for the detection of circulationg antigens. The results were as follows: Mean value of O.D. in control sera was 0.04 (S.D.=0.04). After infection, mean O.D.(S.D) values were changed serially: 0.03(0.01) at 0.5 week(3 days), 0.55(0.50) at 1 week, 0.69(0.45) at 1.5week, 0.20(0.19) at 2 weeks and 0.13(0.10) at 2.5 weeks of infection. They returned, thereafter, to the level before infection. When 0.16(mean+3 S.D.) were considered as cut-off value, those higher than 0.16 were observed only in the sera collected between 1 and 2.5 weeks after infection. Average 8.4 immature worms (2.2 from the lungs and pleural cavities; 6.2 from muscles) were recovered in a rat at 12 weeks after infection. The fact that circulating antigens were not detected after 3 weeks of infection was considered to be caused by the formation of antigen-antibody complexs. PMID- 12886065 TI - Specific IgG antibody responses in experimental cat metagonimiasis. AB - In order to observe the feasibility of serologic diagnosis of metagonimiasis, saline extracts of metacercariae and 4-week old adults were prepared. Sera from 25 experimentally infected cats were collected from 3 days to 12 weeks after infection. Their levels of specific IgG antibody were measured by ELISA together with 3 sera from non-infected cats. Specific IgG antibody levels began to rise in 7 days after infection, reached their peak in 2-4 weeks and made a plateau thereafter. Cats infected with hundreds of adult worms showed minimal rise of the antibody level. Adult antigen was superior to metacercarial antigen in detecting the specific IgG antibody. PMID- 12886066 TI - [The karyotype of Paramphistomum cervi (Zeder, 1790) from Korean cattle] AB - As a series of systematic classification of paramphistomes, the worms in the rumen and reticulum were collected on 214 Korean cattle slaughtered at Jeonju abattoir from January 1986 to April 1987 and were classified by means of morphology. Afterwards, the karyotype of Paramphistomum cervi (Zeder, 1790) was detected by means of modified air-drying method from germ cells of the worms. The results were summarized as follows: In the chromosome number of 254 P. cervi, the haploid cell was n=9 and the diploid 2n=18. The meiotic divisions were observed frequently; 1,924 haploid and 32 diploid cells were reliable. Nine pairs of mitotic chromosomes were homologous in the metaphase stage, and the chromosomes were composed of five medium-sized metacentrics (m), subtelocentrics (st) or submetacentrics (sm) and four small-sized subtelocentrics(st) or submetacentrics(sm). Meiotic metaphase was composed of five medium and four small chromosomes in size. As a series of C-banding method, C-band was showed in centromeric region from all of the haploid germ cells. Whereas chromosome No. 3 and 5 included heterochromatin on the tip region, chromosome No. 4 on the distal region and No. 6 proximal region. And chromosomes No. 2 and 8 showed a remarkable C-band distinguished from other chromosomes. PMID- 12886067 TI - Analysis of antigen specificity using monoclonal and polyclonal antibodies to Cysticercus cellulosae by enzyme-linked immunoelectrotransfer blot technique. AB - To analyse the antigen specificity of patients sera from 24 confirmed neurocysticercosis and a monoclonal antibody, SDS-PAGE using 10-15% linear gradient gel and EITB were done. Cystic fluid, saline extracts of scolex and of whole worm of C. cellulosae, saline extracts of sparganum, hydatid cyst fluid, saline extracts of Fasciola, Clonorchis and Paragonimus were used as antigen. Of protein bands in cystic fluid of C. cellulosae, patient sera reacted frequently to bands of 152, 94, 64, 48, 24, 15, 10 and 7 kDa proteins. To saline extracts of scolex and whole worm of C. cellulosae, patients sera reacted frequently to 94, 64, 52, 39, 34, 15 and 10 kDa bands. Two bands in sparganum extract (130 and 64 kDa) and two bands in hydatid cyst fluid (52 and 27 kDa) were cross-reacting bands with sera from cysticercosis patient. Saline extracts of Fasciola, Clonorchis and Paragonimus did not exhibit cross-reacting bands. Monoclonal antibody to cystic fluid of C. cellulosae was found to react with low molecular weight proteins of 15, 10 and 7 kDa. PMID- 12886068 TI - Histopathological and serological observations on experimental anisakiasis of rabbits. AB - This study was performed to observe histopathological changes and serological reactions in chronic anisakiasis of rabbits. Each rabbit was infected per os with 30 larvae of Anisakis type I. Their sera were collected chronologically and the rabbits were killed for histopathological examination, 3, 13, 20, 30, 60, 90 and 150 days after the infection. The results were summarized as below. Most of the larvae were recovered from the stomach, but a few from the omentum, intestine, mesentery and abdominal wall. The recovery rates and distribution of worms by organ were not differed by duration of infection. Histologically the lesion was abscess type on 13 days, i.e., the dead worms were surrounded by fibrinous exudate, histiocytes and thick zone of numerous inflammatory cells. After 30 days, histiocytes were found to invade the worms and the lesion was changing into abscess-granulomatous type. Also a calcified worm was found on the 30th day. After then the worms were observed to be dissolved slowly until 90 days. On 150 day, only one calcified worm was observed. The levels of serum IgG antibody by ELISA reached their maximum 30 days after the infection. After then, it decreased slowly until 150 days after the infection. Above serological and histopathological findings indicated that antigenic stimulation from degenerating Anisakis larvae was the greatest during the first 30 days after infection. This period was corresponding with the beginning of worm resolution or calcification. Serologic test by ELISA would be a valuable tool for confirming chronic anisakiasis. PMID- 12886069 TI - [Egg detection rates of Enterobius vermicularis in children] AB - The cello-tape anal swab specimens from 936 primary school children in Seoul, 308 primary school children in a rural area and 92 children in Nanjido, were examined for Enterobius vermicularis eggs. Out of 936 specimens from a primary school in Seoul, E. vermicularis eggs were detected in 114 specimens, the egg detection rate being 12.2 percent. In 308 specimens from a primary school in a rural area, 77 (25.0 percent) were positive for E. vermicularis eggs. Among 92 specimens from Nanjido children in Seoul, egg positive rate of E. vermicularis was 28.3 percent. The egg detection rate of primary school children in Seoul was significantly lower than that of primary school children in a rural area and of Nanjido children. The egg detection rate in boys was lower than that of girls. As the age of children increased, the egg detection rate decreased. PMID- 12886070 TI - [A case of calabar swelling suspected as loiasis] AB - This report deals with an imported case of calabar swelling suspective of loiasis, who had been in Nigeria, Africa for 2 years. This 33-year-old Korean technician was admitted to Severance Hospital, Yonsei University Medical Center, because of erythematous swellings on left hand and foot. His peripheral blood showed persistent eosinophilia (over 30 percent of WBC), increased IgE(1,000 unit/ml) and significantly high antifilarial antibody titer with enzyme immunoassay, although no microfilaria was detected on the peripheral blood films. Under the impression of Loa loa infection diethylcarbamazine was administered for a month. Four months later mobile swellings and eosinophilia disappeared, and anti-filarial antibody titers were normalized. It is assumed that the patient had suffered from Loa loa infection, which is the first report on loiasis in Korea. PMID- 12886071 TI - Serological cross-reactivity between Sarcocystis and Toxoplasma in pigs. AB - The development of antibody titers and cross reation between Sarcocystis and Toxoplasma were investigated by means of IFA test and ELISA in pigs experimentally infected with 1.5 x 10(6) S. suicanis sporocysts and 10,000 T. gondii oocysts, respectively. The intact and soluble Sarcocystis antigens were prepared from the bradyzoites harvested by peptic digestion of infected pork. The intact and soluble Toxoplasma, antigens were prepared from the tachyzoites in mouse peritoneal cavity. IgG antibodies in pigs infeded with Sarcocystis and Toxoplasma, respectively were detected first at 2 weeks post infection on both IFA test and ELISA. The antibody titer to Toxoplasma reached its maximum at 6 weeks post infection and decreased thereafter. The antibody titer to Sarcocystis reached its maximum terminally. The cross-reaction titer in pigs infected with Toxoplasma against Sarcocystis antigen was up to 1:16 in IFA test and up to 1:32 in ELISA. The titer in control group was below 1:4 in both reactions. PMID- 12886072 TI - [Immunodepression during experimental Naegleria meningoencephalitis in mice] AB - In order to test the function of lymphocytes in Naegleria fowleri-infected mice, the in vitro blastogenic response of splenocyte cultures to non-specific mitogens was studied. Concanavalin A and lipopolysaccharide stimulation were used as tests of T cell and B cell function. For the first 14 days following N. fowleri infection, lymphoblastic transformation induced by T-cell mitogen was markedly reduced in comparison to the uninfected control mice. The blastogenic response to B-cell mitogen remained depressed in the infected mice up to 14 days after infection. The fluorescent antibody titers of sera of N. fowleri infected mice were between 1:4 and 1:32. The results suggest that there is a suppression of cell mediated immunity during the acute course of experimental Naegleria meningoencephalitis in mice. PMID- 12886073 TI - [Chemotherapeutic efficacy of trimethoprim-sulfamethoxazole (Bactrim) in experimental murine toxoplasmosis] AB - The chemotherapeutic efficacy of trimethoprim-sulfamethoxazole (Bactrim) in mice experimentally infected with Toxoplasma gondii was evaluated. The average survival days and survival rate of mice infected intraperitoneally with 1 x 10(5) trophozoites and treated with Bactrim were compared with those of untreated group. The hematologic findings of blood samples of experimental mice were observed for comparison of side effects between Bactrim and pyrimethamine (Daraprim), the latter of which has been one of the favorable drugs for the treatment of toxoplasmosis. The results are summarized as follows: Bactrim showed a strong evidence of potent anti-Toxoplasma activity. The survival rate of mice administered with 24 mg of Bactrim per mouse per day for 7 days, was 83.3 percent, and the rate was increased to 100 percent in mice administered with two fold concentrated dose of the drug. The average numbers of white blood cells (W.B.C.) in the mouse groups treated with Bactrim or Daraprim were more increased than those only infected with T. gondi. The mice treated with Daraprim, however, showed remarkably decreased numbers of W.B.C. as compared with those treated with Bactrim. The average numbers of red blood cells (R.B.C.) and platelets both in the drug-treated and untreated T. gondii-infected mice were decreased as compared with normal mice. The numbers of R.B.C. in Daraprim-treated mice, however, were more decreased than in Bactrim-treated mice. The average levels of hemoglobin both in the drug-treated and untreated T. gondii-infected mice were decreased, compared with normal mice. But there was no difference in the levels of hemoglobin between Bactrim- and Daraprim-treated groups. In conclusion, trimethoprim-sulfamethoxazole (Bactrim) was proven to be effective and safe for the treatment of murine toxoplasmosis. The efficacy was comparable with pyrimethamine (Daraprim), but bone marrow depression was less severe with Bactrim treatment. PMID- 12886074 TI - [Checklist of mosquitoes (Culicidae) in Korea] AB - A systematic mosquito taxonomic study in Korea was established by Lee in 1971. Since then, many species have been recorded as new to Korea. However, much of this taxonomic information has been distributed in a number of scientific publications. This work attempts to combine this information into a single document, as well as to synthesize revised taxonomic synonyms of species occurring in Korea. Four species, Culex vorax, Culex kangi, Aedes kobayashii and Culiseta kanayamensis listed in our previous paper, are synonyms of Culex halifaxii, Culex jacksoni, Aedes alektorovi and Culiseta bergrothi, respectively. Culex annulus is a misidentified species of Culex pseudovishnui. The subspecies Culex pipiens quinquefasciatus and subgenus Mansonia (Coquillettidia) were treated as full species and genus respectively in a catalog of the mosquitoes of the world. However, not all current authors agree to those specific and generic treatment. Based on available publications the total number of mosquito species recorded in Korea is now 52 representing 9 genera. Forty-five of these 52 are confirmed species with preserved specimen representation. As collection of specimen continues, there is a possibility that representative of 7 unconfirmed species will be found. I hope this paper will provide useful information for entomology workers who are interested in mosquito taxonomy in Korea. PMID- 12886075 TI - Unrecorded Aedes (Edwardsaedes) bekkui (Diptera: Culicidae) from Korea: Notes on taxonomic diagnosis of adult and larva. AB - Thirty-six females and 1 male of Aedes bekkui were captured at human bait in Kyungki Province, Korea Republic, in June 1987. This is the 1st time the species has been recorded from that country. Notes on the taxonomic diagnosis of adults and larvae are given. PMID- 12886076 TI - [Blastogenic responses of splenic lymphocytes to Naegleria fowleri lysates and T cell mitogen in mice with primary amoebic meningoencephalitis] AB - This study was to observe the changes of blastogenic responses of splenic lymphocytes to T-cell mitogens, N. fowleri lysate and concanavalin A, and serum antibody titer during the course of experimental PAM in mice. Naegleria fowleri, strain 0359, was cultured in the CGVS medium axenically and inoculated intranasally with 7 x 10(4) trophozoites for the development of experimental PAM in mice. The amoebae were subjected to ultrasonication and centrifuged at 20,000 g for 60 minutes, and filtered through 0.2 micro-m filter membrane. The supernatant, N. fowleri lysate, was used as T-cell mitogen, and antigen for ELISA. The serum antibody was examined by ELISA using peroxidase conjugate. Two hundred micro-l of 10(6) splenocytes in RPMI 1640 containing 10% fetal calf serum were added to each well of a microtiter plate. To each well was added T-cell mitogens, 100 micro-g/ml of N. fowleri lysate or 4 micro-g/ml of con. A, and the plates were incubated for 42 hours at 37 C in 5% CO(2) incubator. Cultures were pulsed with 1 micro-Ci of methyl-(3H)-thymidine 6 hour before harvesting. The mean blastogenic response of the splenocytes to N. fowleri lysate was reduced, whereas that to con. A was also reduced up to on day 11 after infection. Both of these results were statistically significant compared with those of uninfected control group. The serum antibody titers were increased gradually up to day 15. The results indicated that there was an impairment of the blastogenic response of splenocytes to N. fowleri lysate during the acute course of experimental PAM in mice. PMID- 12886077 TI - [Detection of IgG antibodies with immunofluorescent antibody technique in human trichomoniasis] AB - The indirect fluorescent antibody(IFA) test was used to detect serum IgG and IgM antibodies to Trichomonas vaginalis in 31 vaginal trichomoniasis, 7 candidiasis and in 20 non-infected healthy wonem with antigen prepared from axenic culture of Trichomonas vaginalis isolated from vulvovaginitis patient. The results were as follows: In 31 vaginal trichomoniasis the positive reactions of IgG antibody were 27 in the 1/8 dilution or higher and 4 in the 1/4 dilution whereas in healthy women the reaction showed signigicantly low as in the 1/4 dilution of below. The sensitivity and specificity of IFA test for IgG antibody to trichomonad antigen in this study were 87.1% and 100%, respectively. No significant difference of IgM antibody levels between vaginal trichomoniasis and healthy women was observed. No relation between the levels of IgG and IgM antibodies to trichomonad antigen by IFA test was observed. No relation between the time lapse and the level of serum IgG antibodies in IFA test of vaginal trichomoniasis was regarded. In conclusion the present study suggests that IFA test in trichomoniasis could be a useful tool for detection of anti-trichomonad IgG antibodies and applicable as an immundiagnostic method. PMID- 12886078 TI - Toxoplasma antibodies by indirect latex agglutination tests in zoo animals. AB - Total of 216 animals conserved in Seoul Grand Park were examined on the antibody titers of Toxoplasma by the indirect latex agglutination test, 20 out of 131 cases (15.3%) in mammals, 2 out of 75 cases (2.7%) in birds, and none in reptiles, according to species, 15 out of 68 species (22.1%) in mammals, 2 out of 36 species (5.6%) in birds showed positive antibody titers when the titers of 1:32 or higher were regarded as positive. In mammals, it appeared as positive in 2 out of 6 cases (1 out of 3 species ) in marsupials, 1 out of 15 cases (1 out of 11 species) in primates, 1 out of 1 case in bats, 6 out of 13 cases (5 out of 10 species) in carnivores, 1 in 12 cases (1 species out of 3) in odd-toed ungulates, 9 out of 80 cases (6 species out of 38) in even-toed ungulates, and none in rodents and in whales. In birds, 1 out of 21 cases (1 out of 7 species) in gallinaceous birds and 1 out of 6 (5 species ) in parrots appeared to have the positive antibody titers of Toxoplasma. And, none of reptiles showed positive. Frequencies of positive antibody titers were high in 1: 64, 9 cases in mammals followed by 1: 32, 6 cases, 1: 128, 3 cases, and 1: 256, 2 cases, respectively. Two positive cases in birds appeared to be 1: 64. PMID- 12886079 TI - In vitro effect of praziquantel on Paragonimus westermani by light and scanning electron microscopic observation. AB - The effect of praziquantel on P. westermani exposed in vitro was observed by stereomicroscope, light microscope and scanning electron microscope. Following results were found. The worms incubated in 0.01 micro-g/ml praziquantel were moving after 26-hour incubation. However, all of them were immobilized immediately after incubation in solutions over 0.1 micro-g/ml concentration. All of the exposed worms showed severe vacuolization not only in tegument but in subtegument, intestine, ovary, testis, Mehlis' gland and excretory bladder. Vacuoles in tegument burst out to form craters. As incubation time went on, tegumental structure was disintegrated severely. The worms exposed to praziquantel were observed to be immobilized and be vacuolized of all tissues. Disintegration of reproductive organs suggests that praziquantel have suppressive effect on egg production when the flukes are not killed. The drug effects were found more related with incubation time than with drug concentration. PMID- 12886080 TI - [Karyotype of Fasciola sp. obtained from Korean cattle] AB - As a series of systematic classification for Korean common liver fluke, Fasciola sp., karyotype was investigated by means of the modified air-drying technique and of the regular Giemsa staining, Also, C-staining method was applied for detailed karyological analysis from the germ cells of the fluke. The following is a brief summary of the leading facts gained through the experiment. Korean Fasciola sp. was classified into three types based on their chromosomal complements; individuals with 20 or 30 chromosomes and with a 20/30 mosaic constitution. Worms having 30 chromosomes represent a triploid form with 3 sets of 10 basic chromosomes, while those with 20 chromosomes were diploid and mosaic individuals were 2n/3n mixoploid. The frequency of the individual type calculated is as follows; 67.45% of 212 flukes examined was of diploid, 10.85%, triploid, and the rest, 21.7%, mixoploid, respectively. In many cases, two or three types were found in the peculiar bovine host while single type inhabitant was about 20% out of 52 cases. The twenty chromosomes consisted of 1 parir of large metacentrics, 4 pairs of medium-sized subtelocentrics, and 5 pairs of small submetacentrics, while constitution of the thirty chromosomes was nearly interpreted as a triploid form with 3 sets of 10 basic chromosomes. The high centromeric indexes of both types are the first pairs among all the examined, and 37.93% was of diploid and 47.93%, triploid, respectively. In mixoploid individuals, constitution of the chromosomes of diploid or triploid cells was the same as that of diploid or triploid individuals. All the chromosomes of the germ cells in both types showed C-band around the centromeic region and especially the chromosomes no's 3, 7, and 8 showed a remarkable C-band distinguished from other chromosomes. The variance for the sizes of the worms and the eggs were not parallel with three different genotypes in Korean common liver fluke. PMID- 12886081 TI - [Passive transfer of immunity against Clonorchis sinensis by peritoneal exudate cells in mice] AB - This study was undertaken to evaluate the role of peritoneal exudate cells in the transfer of immunity against the liver fluke, Clonorchis sinensis in the inbred BALB/c mice. Ten donor mice were divided into 2 groups. One group consisted of 5 mice was infected orally with 20 metacercariae of C. sinensis, and the other group was injected intraperitoneally with 20 excysted larvae. Thirty days after immunization, the peritoneal exudate cells were obtained from the donor mice. Twenty recipient mice were divided into 4 equal groups for the purpose of primary immunization. The mice of Group I were injected intraperitoneally with 2 x 10(6) peritoneal exudate cells of the donor mice infected orally, those of Group III were injected intraperitoneally with 2 x 10(6) peritoneal exudate cells of the donor mice injected intraperitoneally. Those of Group II were injected orally with 20 metacercariae of C. sinensis. The group IV mice served as controls. Four days after the primary immunization all recipient mice were challenged orally with 20 metacercariae of C. sinensis, and then killed 30 days after the challenging infection. When the peritoneal exudate cells were injected into the recipient mice, pronounced reduction in eggs per gram of the feces was found in the mice of Group I and Group II, but no reduction in those of Group III. In the worm burdens of C. sinensis, the number of flukes found in the mice of Group II was only significantly less than those in the control group(IV). In addition the number of plaque forming cells per spleen in the mice of Group II was found larger than those in Group I. It is likely that donor peritoneal exudate cells transferred to the recipients might result in the production of relative immunity. PMID- 12886082 TI - Scanning electron microscopical findings of Echinochasmus japonicus tegument. AB - Tegumental ultrastructures of Echinochasmus japonicus were observed by scanning electron microscopy. The worms were recovered from albino rats which were experimentally infected with the metacercariae obtained from Pseudorasbora parva. Followings are summarized findings. The worms were minute and plumpy gourd-shaped with attenuated anterior and round posterior end. The tegument of whole body was wrinkled transversely and covered with cobblestone-like cytoplasmic processes. Head crown was armed with 24 collar spines which were embedded in cytoplasmic pockets. The spines were arranged in a row with an interruption at dorsomedian line, however, the 2nd and the 4th spines were outstretched more than others. Oral and ventral suckers were muscular with numerous type II sensory papillae, and genital pore opened between the two suckers. Tegumental spines were spade shaped with broad base and pointed tip. They were compact in ventro-lateral tegument or dorsal surface of anterior body. The were not found between the two suckers and dorsal surface of posterior body. Two types of sensory papillae, uni ciliated (Type I) and roundly swollen sensory papillae (Type II), were observed. The type I papillae were chiefly distributed on ventral surface of tegument and type II were on the lips of suckers. Arrangement of collar spines, shape and distribution of tegumental spines or sensory papillae are regarded as characteristic features of E. japonicus. PMID- 12886083 TI - [Epidemiological survey on Metagonimus yokogawai infection in the eastern coast area of Kangwon-province, Korea] AB - An epidemiological survey on Metagonimus yokogawai infection was performed in the eastern coast area of Kangwon province. Formalin-ether concentration technique and Stoll's dilution egg counting method were employed to figure out the prevalence and intensity of M. yokogawai infection and eating habits of inhabitants with raw freshwater fish. The results are summarized as follows: The prevalence rate of M. yokogawai infection was 6.6% among a total of 2,357 examinees; 8.6% (115 out of 1,345) in male and 4.0% (40 out of 1,012) in female. The prevalence rates were different by the streams where the examinees were residing: 17.8% along the area of Ohsip-cheon (stream), 11.8% along Maeup-cheon, 1.7% along Hwasang-cheon, 1.3% along Yeongok-cheon, 0.9% along Namdae-cheon, 0.9% along Sa-cheon and 0% in the other streams (Yongchon-cheon, Kangnung Namdae-cheon and Jusoo-cheon) in decreasing order. By social strata, the positive rate was 8.0% (122 out of 1,521) in general inhabitants and 3.9% (33 out of 836) in school children. By the intensity of infection by means of EPG counts in feces; the light infection (less than 400/EPG) was 74.1%, moderate infection (401-1,000/EPG) 17.3% and heavy infection (more than 1,001/EPG) 8.6%. Twelves out of 50 sweetfish (Plecoglossus altivelis) (24%) caught from the streams in eastern coast of Kangwon-Do were infected with metacercariae of M. yokogawai, and the number of larvae detected were 70.7 per fish in average. The metacercarial infection rate in fish were also different by the streams; 100% in Ohsip-cheon of Samcheok-Gun, 14.3% in Sa-cheon, 16.7% in Yeongok-cheon of Myeongju-Gun and 0% in Namdae-cheon of Yangyang-Gun, respectively. Six hundred ninety five (695) out of 1,396 inhabitants and school children (49.8%) were experienced in eating raw freshwater fishes (Plecoglossus altivelis, Tribolodon hakonensis, Coreoperca sp., Moroco sp., Carassius sp. & Cyprinus sp. and Zacco platypus). The data were assayed with the questionnaire collected from the examinees. In summarizing the above results, it is known that the Ohsip-cheon area of Samcheok-Gun is highly infected region, and the other areas of Kangnung Namdae-cheon, Yeongok-cheon and Yangyang Namdae cheon are very lowly infected or non-infected regions for M. yokogawai infection. PMID- 12886084 TI - [Studies on the Metagonimus fluke in the Daecheong reservoir and the upper stream of Geum river, Korea] AB - The prevalences of the fluke belonging to genus Metagonimus have been reported along the upper stream of inhabitants by several workers since 1980, however the taxonomical problems of the fluke was not yet settled. The larval flukes; cercaria and metacercaria as well as their intermediate hosts, and adult were studied in order to identify the Metagonimus in the areas. The results obtained are summarized as follows: The snails, Semisulcospira globus were collected from the three different localities along the upper stream of the River. The cercariae were found from 125(7.2%) out of 1,730 snails by natural emerging method, and were identified into 5 species including Metagonimus sp. (3.7%), Pseudexorchis major(1.4%), Cercaria nipponensis (0.9), Cercaria incerta (0.6%) and Cercaria yoshidae(0.6%). Cercariae of Metagonimus species had four to five oral spines on its anterior of the first line. The cercariae of Metagonimus were experimentally exposed to goldfish. Infection rate was 22.9% out of 105 goldfish, and the encysted metacercariae were found in fins(86.7%) and on scales (13.7%) of the fishes, but not in their muscle, head or visceral organs. Seven species of fish were caught in the Daecheong reservoir and the upper stream. Infestations with metacercaria of Metagonimus were found 100% in Opsariichtys bidens and the parasitized numbers of the metacercariae were observed from 250 to 2,400 per fish. In the upper stream, Zacco temmincki, Z. platypus and Pseudogobio esocinus were infected 100% with the metacercaria, on the other hand, the fishes caught in the reservoir showed the lower infestation rates, and a few metacercariae found in the fishes Carassius carassius and Cyprinus carpio in the reservoir and the stream. The majority of metacercariae was detected only on the scales of fishes. In order to know the infectivity and the distribution patterns in the intestine of hosts, rats and dogs were infected with the metacercariae obtained from O. bidens and Z. platypus. In addition the metacercariae obtained from Z. temmincki, P. esocinus and goldfish were given to the rats. The recovery rates of the worms in the small intestine of dogs were higher (63.3-65.8%) than those of the rats (3.5-31.6%). The flukes were found mostly in the middle and the lower part of small tntestine of the rats and the dogs, but no worm was collected in the upper part of the intestine of rats. The size of adult flukes varied by the hosts. In the adult flukes, oral sucker was smaller than ventral sucker, and the right and left testes were located diagonally, the uterine tubules circled around the upper left testis. The average egg size was 29.1 x 17.7 micro-meter. According to the above results, the flukes belonging to genus Metagonimus distributed along the Geum River was concluded to be identical with Miyata type of M. yokogawai as that Saito had proposed. PMID- 12886085 TI - A case of human thelaziasis in Korea. AB - The 15th case of human thelaziasis is reported from Korea. The patient was a 15 year-old girl from Seoul. Both of the worms which were removed from her eye were female and were identified by the number of transverse cuticular striations (281, 172 and 162/mm in the oesophageal, middle and posterior portions, respectively), as Thelazia callipaeda. The patient owned a pet dog ADDITIONAL ABSTRACT: The 15 year-old patient had herself removed a worm from one eye. Two worms were identified morphologically as Thelazia callipaeda. The source of infection was uncertain, but the patient was known to have close contact with a dog. The authors refer to 14 previously reported cases of thelaziasis in Korea. PMID- 12886086 TI - [Head louse infestation among school children in Sanbuk-myon, Mungyong-gun, Kyongsangbuk-do (1985)] AB - In order to know the prevalence of head louse among the school children in rural area, we visited 3 primary schools and a kindergarten in Sanbuk-myon, Mungyong gun, Kyongsangbuk-do in July 1985, and examined the children to identify the adult worms of head louse and/or its nits on the hair. The overall infestation rate was 91.9% among 420 children examined, and there was a difference between schools (P< 0.05). the school girls' rate, 96.1% was higher than that of bods' 88.9%(P< 0.05). By grade, the rate of infant school was 88.5% and 1st to 6th grade were 100, 100, 95.8, 90.4 89.7, 80.0% respectively. Above result indicates that the head louse is an important health problem in this community. Therefore, the chemotherapy and the effort to improve the hygiene are necessary in this community by physicians, teachers and health administrators. PMID- 12886087 TI - Growth and development of Fibricola seoulensis metacercariae in tadpoles. AB - In order to observe the growth and development of Fibricola seoulensis metacercariae, the tadpoles of Rana nigromagulata were experimentally infected with the cercariae. The metacercariae of various developmental stages were recovered from the tadpoles after 2 to 65 days of infection. They were prepared for morphological observation, and were given orally to mice to observe their infectivity. The following results were obtained. All of the tadpoles exposed to the cercariae were observed to harbour the larvae in their abdominal cavity. The young metacercariae of 2 days after infection were 121.1 micrometer long and 63.3 micrometer wide. They grow linearly for the first 14 days to be 262.0 micrometer long and 166.4 micrometer wide. Thereafter, no more growth recognized until 65 days. The larvae of 2 days old were similar with cercarial body and had 2 suckers, a pharynx, two ceca and a primordium of germ cells but no tribocytic organ. On the 8th day, they had tribocytic organ, and their morphology resembled that of mature metacercariae. The metacercariae younger than 10 days could not infect the mice. Only the metacercariae older than 14 days had infectivity. The recovery rate increased by the age of metacercariae from 19.0% in 14 days old to 70.0% in 40 days old. Above findings indicate that the tadpole is indispensible for metacercarial development and it needs at least 2 weeks for maturation. The tadpole is a pivotal host in the life cycle of F. seoulensis for connection between the snail and the frog. PMID- 12886088 TI - The effect of temperature and salinity on maturation and hatching of Fibricola seoulensis eggs. AB - This study was carried out to reveal the effect of temperature, salinity and aeration on maturation and hatching of Fibricola seoulensis eggs. The eggs were incubated and were observed daily for the appearance of eyespots and hatching. The results were summerized as follows: From 4-5 days after incubation in distilled water at 28C or at 11-26C, the eyespots began to appear and the rate of eggs with eyespots were over 90% in 28C on the 7th or 8th day. However, eyespots did not appear in 5-15C or 4C by the 18th day. The mature eggs began to hatch at the 8th day, and hatching rate 2 weeks after incubation was over 90% at 28C, but it was below 5% at 11-26C, and 0% at 5-15C and 4C. Aeration did not influence the appearance of eyespots nor hatching. In saline under 0.6%, the rates of eyespots were over 90% on the 7th day. The rate was 55.0% in 0.9% at 20 days, and 0% in 1.2%. The hatching rates in salines below 0.3% concentration were over 90% by 14 days of incubation. However, the rate decreased to 44% in 0.6% saline and to 0% over 0.9% salinity. The eggs incubated in the dark hatched in 12.5% on the 10th day, but hatching rate of maturation eggs increased to 85.7% within 2 hours after exposure to light. Above results demonstrated that the best temperature for maturation and hatching of F. seoulensis eggs was 28C, and the miracidia began to hatch at 8-9 days after incubation. In the field hatching and invasion into snails of the miracidia may occur from May to September in Korea. In salines under 0.3 percent concentration maturation and hatching were not influenced, but as salinity increased hatching was inhibited more than maturation was. PMID- 12886089 TI - [Experimental and epidemiological studies on the life cycle of Echinostoma hortense Asada, 1926 (Trematoda: Echinostomatidae)] AB - Recently there have been some reports on human infections of Echinostoma hortense in Korea. It was found that a few species of freshwater fishes were playing the role of the second intermediate host of E. hortense. However, molluscan intermediate host has not been identified yet in Korea. The present study aimed to establish the life cycle of E. hortense in laboratory. Experimental studies such as egg production from the rat, development of the eggs in vitro, exposure of miracidia to freshwater snails, shedding pattern of cercariae from infected snails, morphology of cercariae, cercarial infection to the second intermediate host and infection of metacercariae to the difinitive hosts were done. In addition, epidemiological surveys on the infection status in inhabitants and house rats, and on the natural infection of larval echinostomes in the snails and fishes were carried out along the South Hangang-river. The results obtained were as follows: The eggs deposited from adults in physiological saline were cultivated at room temperature (20-24C). The miracidia were firstly observed on 8 days after cultivation, and 85.5 per cent of the eggs contained the mature miracidia on 11 days after cultivation. More than 90 per cent formed the miracidia when cultivated at temperature 22-27C. Hatching of the miracidia began on 12 days after cultivation and continued for a week. The size of the miracidia was 103.0 x 51.4 micrometer in average. The motility of miracidia were active up to 8 hours after shedding, but they were all dead within 10 hours after shedding. A freshwater snail, Radix auricularia coreana was cultivated in aquaria. A hatched F1 snails from the egg masses were exposed to 20 miracidia respectively. Escape of cercariae started on 15 days after infection. Radix auricularia coreana was experimentally identified as the first intermediate host of E. hortense in Korea. Cercarial shedding started on 15-20 days after infection by snail, continued for about 10 days (8.8 days in average). Infected snails were dead within 32 days after the miracidial infection. About 1,335 cercariae (328-1,994) per snail were shed in its life, and 119 cercariae in average per snail per day were shed. The cercariae were motile for more than 24 hours, and then squirming at the bottom until death. The body and tail sizes of cercariae were 356 x 186 micrometer and 510 x 68 micrometer in average, respectively. The rediae parasitized in the snail hosts were found mainly around the pericardial regions, and their size was 1,575 x 258 micrometer in average. The numbers of developing cercariae in a mature redia were 14 in average (7-20 in range). The numbers of rediae in a snail were 102 in average on 15 days after miracidial infection and 221 in average on 28 days. Three uninfected Misgurnus anguillicaudatus, less than 6.5 cm long were used in for the cercaria1 infection. They were all exposed with 755 cercariae, and examined at 5-day intervals starting from 10 days after infection. All the fihes were infected with metacercariae of E. hortense and a total of 275 was found infected (36.4 per cent). The metacercariae were fed to rats and the adult worms were obtained on 15 days after infection. The infected rats began to deposit the eggs on 11 days after infection. The number of eggs deposited per day per worm (EPD/worm) was 400-500 on 3 weeks after infection and was increased to 1,000-1,500 on 4 to 17 weeks, then decreased to 800 on 2l weeks after infection. A total of 745 stool specimens collected from 576 male and 169 female residents of 8 different villages along South Hangang basin was examined. Out of 745 specimens, the eggs of Echinostoma sp. were found in 2 cases (0.3 per cent). Of 34 house rats one showed egg-positive (2.9 per cent). Total 971 Radix auricularia coreana collected from 7 sampling stations were examined for shedding of cercariae. Three snails (0.3 per cent) shed the cercariae of E. hortense. A total of 119 out of 542 freshwater fishes (22.0 per cent) had the metacercariae of E. hortense. The fishes parasitized with the metacercariae were 4 out of 14 examined species. The infection rate of 4 species were 34.1 per cent (106 out of 311) in Misgurnus anguillicaudatus, 30.4 per cent 7 out of 23) in Misgurnus mizolepis, 4.3 per cent (2 out of 46) in Moroco oxycephalus and 22.2 per cent (4 out of 18) in Odontobutis obscura interrupta. In summarizing the above results, the first intermediate host of E. hortense was found as Radix auricularia coreana in Korea. Also it took about 46 days for the shortest completion of a life cycle of E. hortense in summer; that is, 10 days for miracidial development in eggs, 15 days for cercarial development in the snail, about 10 days for metacercarial development in the second intermediate hosts, 11 days for the maturation as the adults in the definitive hosts. The natural infection rates of E. hortense in the intermediate hosts were relatively high but those in the definitive hosts were low in the middle areas of South Hangang basin. PMID- 12886090 TI - [Studies on beta-glucuronidase activities in liver, stomach and small intestinal tissues of rabbits infected with Clonorchis sinensis] AB - The author has studied the beta-glucuronidase activity in several tissues such as liver, stomach and small intestine of the male and female rabbits infected with different doses of metacercariae of Clonorchis sinensis. The metacercariae of Clonorchis sinensis were isolated from Pseudorasbora parva caught in Kim Hae by digestion technic. The experimental animals were sacrificed in the period of 1, 7, 14, 21, 28 and 35th days following the infection. The results obtained were summarized as follows: In the groups infected with 100 metacercariae, beta glucuronidase activity was slightly increased during the entire periods than control rabbits. It was the highest in the first day with 1.535 and 1.421 mu/g, 14th days with 2.521 and 2.200 mu/g, and then lowered by the time, gradually. In the groups infected with 500 metacercariae, beta-glucuronidase activity was highly increased on the first day with 1.535 and 1.866 mu/g than that 100 metacercariae groups according to each organs. It was the highest on the 7th day and 14th day. In the groups infected with 1,000 metacercariae, beta-glucuronidase activity was remarkably increased in the first and 14th days according to each organs, and then lowered gradually day by day. beta-glucuronidase activity of all organs was more increased than that of normal organs and the highest activity in the liver with 2.521 mu/g, intestine(1.612) and stomach (1.581) respectively. beta-glucuronidase activity of rabbits was higher in the female than in the male. On the basis of these results, it was suggested that beta-glucuronidase activity was affected by the duration of infection and by the number of Clonorchis sinensis, according to the organs and sex of the rabbits. PMID- 12886091 TI - Purification of cystic fluid antigen of Taenia solium metacestodes by affinity chromatography using monoclonal antibody and its antigenic characterization. AB - This study was undertaken to purify cystic fluid (CF) antigen of Taenia solium metacestodes by affinity chromatography using specific monoclonal antibody (McAb) and to characterize the antigenicity of the purified antigen. The hybridoma cell lines, prepared by fusion between mouse plasmacytoma and spleen cells from BALB/c mice immunized with CF, secrected antibodies reacting to various helminthic antigens. Majority of cell lines reacted to CF only but some also reacted to parenchymal antigen of T. solium metacestodes, adult T. saginata, sparganum, hydatid cystic fluid, Paragonimus westermani and Clonorchis sinensis, either in combination with CF, other antigens or independently. Cloned cells derived from monoclonal lines also produced antibodies reacting either to CF only or to other helminthes in combination or independently. These results indicated that CF of T. solium metacestodes contained proteins which possessed antigenic determinants not only specific to CF but also cross reactive with the afore-mentioned helminthes. CF of T. solium metacestodes was purified by affinity chromatography using the McAb which reacted to CF and parenchymal antigens. The affinity-purified antigen(A-Ag) and unbound pool(U-Ag) were separated. A-Ag showed 2 protein bands by disc-PAGE whereas CF exhibited 6 bands and U-Ag consisted of all bands CF had. The diagnostic significance of A-Ag was evaluated by ELISA in human neurocysticercosis and other helminthic and neurologic diseases. By A-Ag, the levels of the specific IgG antibody, as shown by absorbance in sera and CSF, were lower than those of CF and U-Ag. Accordingly, the sensitiviy was about 70 % of CF and U-Ag. However, the nonspecific positive reactions to CF and U-Ag, observed in sparganosis, T. saginata infection and paragonimiasis did not occur when A-Ag was used. These results indicated that the affinity-purified A-Ag had the higher specificity but the lower sensitivity as a diagnostic antigen in cysticercosis, probably because it only detected a single or limited numbers of monospecific antibodies among the diverse polyclonal antibodies produced in the patients with neurocysticercosis. PMID- 12886092 TI - Serologic follow-up study in neurocysticercosis patients by ELISA after praziquantel treatment. AB - /A total of 69 patients of confirmed neurocysticercosis was followed serologically by ELISA up to 22 months after praziquantel treatment. The intervals and numbers of follow-up were variable by patients. Serially collected samples of serum and CSF were examined simultaneously for their specific IgG antibody levels by ELISA, using cystic fluid, saline extracts of bladder wall and scolex as antigen. Within 4 months after praziquantel treatment, the antibody levels were elevated temporarily in both serum and CSF in most patients. In some cases antibody levels exhibited steady declining tendency after the treatment. Concomitant administration of dexamethasone appeared to suppress the elevation of antibody levels. The rate of mean absorbance of antibody changed more in serum than in CSF. The rate of elevation was greater in antibodies to parenchymal antigens than that to cystic fluid, but absolute difference of antibody levels was greater in anitbody to cystic fluid. Previously negative samples for IgG antibody may become positive after praziquantel treatment, which could be used as a complementary tool(provocation test) in serodiagnosis. One month was considered to be sufficient interval for the follow-up test for that purpose. In the follow up of up to 22 months, only few cases of chronic neurocysticercosis showed declining tendency of IgG antibody levels below negative range. During acute encephalitic attacks in chronic patients, IgG antibody to parenchymal antigen were elevated in CSF temporarily. These results indicated that serologic follow up of every year was recommendable to differentiate the cured patients from chronic patients with slowly calcifying lesions. PMID- 12886093 TI - [Fractionation of antigen for ELISA of bovine fascioliasis] AB - In order to obtain the most specific and sensitive antigen from crude antigens of Fasciola hepatica for the immunodiagnosis of bovine fascioliasis by the enzyme linked immunosorbent assay(ELISA), phosphate buffered saline extract of F. hepatica was prepared. The crude extract was fractionated into 7 antigens using Sephadex G-100 column chromatography. Seven fractionated antigens were applied to ELISA, precipitation test and intradermal test, respectively. Results obtained are as follows: The specificity (95 per cent confidence interval in negative sera of bovine fascioliasis; Mean+2 x SD of absorbance ) of the first (MW>150,000) and the second antigens (MW 120,000) were 93.7 per cent, but those of others including crude antigen showed 100 per cen.t. The sensitivity (positive sera of bovine fascioliasis having higher values with compared to the criterion) of the first, the sixth (MW 16,000) and the seventh antigen (MW<5,000) were 91.6 per cent, 87.5 per cent and 0 per cent, respectively, but those of others showed all 100 per cent. The absorbance by ELISA using the fifth antigen (MW 26,000) was 8.43-folds higher in the positive sera than that in the negative sera. This could be used as one of the most specific antigens for the immunodiagnosis of bovine fascioliasis. In Ouchterlony test, precipitin lines were not found in the sera naturally infected with F. hepatica, but some were found in the sera of rabbits immunized with the crude antigens. The numbers of precipitin lines in the sera of rabbits were different in the different fractionated antigens. They were 6 in the crude, 2 in the second and the third antigens, 1 in the forth, the fifth and the sixth antigens and absent in the seventh antigen, respectively. The wheal size for the bovine infected with F. hepatica was 2.46+/-0.15 cm in the intradermal test antigen (saline extract of F. hepatica) supplied by the Veterinary Research Institute, Rural Development Administration, Korea. The wheal size of the first, the second and the third antigens were larger than that of intradermal test antigen, whereas those of the forth, the fifth, the sixth and the seventh antigens showed smaller than that of the intradermal test antigen. The results suggest that the fifth antigen may be specific antigen for the immunodiagnosis of bovine fascioliasis. PMID- 12886094 TI - [Purification of antigenic proteins of Paragonimus westermani and their applicability to experimental cat paragonimiasis] AB - This study was designed to evaluate the partially purified antigens which were fractionated from crude extract of Paragonimus westermani and to monitor the enzyme-linked immunosorbent assay (ELISA) in experimental cat paragonimiasis during the course of infection as well as before and after chemotherapy. Crude extract of 6-month-old adult P. westermani was fractionated to 5 antigens by successive applications of ammonium sulfate precipitation, ion exchange chromatography and gel filtration. And the cats, 10 in each group, were infected with 60, 30, 15, and 5 metacercariae, then the half of each group was treated with praziquantel 2 times in one day of 100 mg per kilogram of weight on 150 days after the infection. Sera were collected every 10 days. ELISA was performed with the concentration of 2 microgram/ml antigen, 100 times diluted sera and 1,000 times diluted alkaline phosphatase conjugated anti-cat IgG. The results were as follows: Absorbance by ELISA with proteins precipitated by differential concentration of ammonium sulfate was the highest at 51-65 per cent precipitate (PA2), followed by 0-50 per cent precipitate (PA1), 66-80 per cent precipitate (PA3), and 81-90 precipitate (PA4). Unprecipitated protein over 90 per cent ammonium sulfate (PA5) showed the lowest antigenicity. Fractionation of PA1, PA2, and PA3 through the DEAE-cellulose column did not differentiate the antigenic proteins. By passing through the Sephadex G-200 column, PAl and PA2 were fractionated to high molecular weight proteins and those of low molecular weight which showed high absorbance by ELISA (PA1-I, II and PA2- I, II). But PA3 was shown to have a fraction of high molecular weight proteins (PA3-I) which showed high antigenicity. SDS-polyacrylamide gel electrophoresis of PA1-I, PA1-II, PA2 I, PA2-II, PA3-I, and crude extract was performed. Fraction PA1-I was composed of proteins which had the molecular weight of 270 kilodaltons (KD) to 196 KD; of them 220 KD protein was major band. Fraction PA2- I was composed of 255-225 KD, and PA3-I, 255-240 KD, respectively. Fraction PA1-II and fraction PA2-II consisted of 30 KD proteins. Absorbance by ELISA began to increase within 10-20 days after the infection and reached the highest on 140-180 days, then made plateau thereafter. Absorbance by ELISA decreased after praziquantel treatment. In 60 metacercariae infection group, the absorbance had been decreasing, but remained within the positive range during observation period, while those of 30, 15, and 5 metacercariae infection groups turned to negative range. Fraction PA1 II showed the highest antigenicity in ELISA, then fraction PA2-I, fraction PA1-I, fraction PA2-II, fraction PA3-I and crude extract followed. In early phase of infection, the absorbance of fraction PA1-II showed more rapid increase than those of the other fractions and it came to positive range at 20-30 days after infection. PMID- 12886095 TI - [ELISA of paragonimiasis in cat by crude and purified antigens of Paragonimus westermani] AB - Enzyme-linked immunosorbent assay (ELISA) using crude and affinity-purified antigens of adult worms of Paragonimus westermani was performed for infected cat sera with different worm burden, from preinfection to 18th week after infection. Crude antigen was used with supernatant of homogenated worms by freezing-thawing method, and the supernate was centrifuged for l hour at 10,000 rpm at 4C. Affinity-purified antigen (antibody-bound antigen) was prepared from fractions (bound and unbound) of crude antigen by affinity chromatography on CNBr-activated sepharose 4B, and IgG as a ligand was prepared from paragonimiasis cat serum (6 months infected) obtained by ammonium sulfate (40-45 per cent saturated) precipitation method. By SDS-PAGE, crude antigen showed 22 polypeptide fractions while purified antigen showed 4 fractions: 36, 400, 34,700,27,600 and 11,500 in molecular weights. All cats were divided into five groups(G1-G5) by different worm burdens. The mean of recovered worms (+/-SD) and the number of cats in each group are as follows:G1, 2 worms(0) and 4 cats; G2, 4.75(+/-0.66) and eight; G3, 10.75(+/-1.92) and four; G4, 25.20(+/-3.43) and five; G5, 48(+/-12.63) and five cats. The results were summarized as follows: The antibody levels(OD value) increased by worm burden in G1 to G4 generally. However, individual antibody levels were not exactly related with worm burden in all groups, especially there was a wide difference in G4 and G5. These results suggested that the worm burden in G4 (about 20 - 30 worms) is enough to produce antibody maximum in cats of 2~3 kg weight. The antibody levels increased significantly (p<0.05) compared to control sera at the 3rd week in G1 and G2, at the 2nd week in G3, and at the 1st week in G4 and G5. Especially in the 4th week, OD value increased more in G1(p<0.001) and in G2 to G5(p<0.01). In the pattern of antibody levels by ELISA in each group, OD in G1 increased to the 18th week continuously, in G2 OD was maintained same after the 16th week, but in G3 it decresed after the 16th week, and it was maintained same in G4 and G5 after the 14th week. The antibody levels by ELISA with the affinity-purified antigen were higher than those with crude antigen in all groups generally. Especially, the difference of OD values between two antigens was larger from the 4th to the 10th week. In G1 and G2 OD with purified antigen was higher than that with crude one to the 18th week. It was also higher in G3 than that with crude antigen to the 16th week and OD of G4 and G5 were higher before the 14th week than that with crude antigen, however became lower at the 16th week. Consequently, the antibody level in ELISA with affinity purified antigen was more sensitive at the early weeks after infection and in light infection groups than that with crude antigen. PMID- 12886096 TI - Infestation status of Paragonimus westermani metacercariae in the second intermediate host in Ulchin county, Kyungpook Province. AB - A study on the population density of crayfish intermediate hosts and infestation status of crayfish with encysted larvae of Paragonimus westermani in Ulchin county, Kyungpook province, Korea was conducted from May to October in 1986. The population density of the crayfish ranged from 1 to 13, with an average of 4 per manhour. Among the six habitats, Ducheon had a somewhat higher density than that of the others. Of eight hundred and seventeen crayfish examined, 127 or 15.5 per cent harboured the metacercarial larvae of Paragonimus westermani. The majority of the larvae were found in three parts of the body: most frequently in the cephalothorax, next in the gills, and then in the liver. The average number of metacercarial larvae per infected crayfish ranged from 1.0 to 1.9, with an average of 1.7. Summarizing the results, this study indicates that the population density of crayfish intermediate host and infestation rates for the crayfish with encysted larvae of Paragonimus westermani in Ulchin county of Kyungpook province is relatively high. PMID- 12886097 TI - [Efrect of diethylcarbamazine against Brugia malayi infection on Cheju Island, evaluated in 1965] AB - A study on the effect of diethylcarbamazine (DEC) (Supatonin) against Brugia malayi infection was conducted on Cheju Island in September 1965. A total of 182 persons living in a village of Aiwol Myun, Bukcheju-Gun was examined for microfilaraemia. Microscopic examination of smears of 20 microliter of blood revealed a microfilaria positivity rate of 28.5 per cent. At the end of September 1965, 34 confirmed microfilaria positive cases were treated with DEC at a daily dosage of 5 mg/kg body weight. A full course of 12 days of drug administration divided of two rounds for 6 days each was used. The first round of treatment was given under a strict supervision of the author in order to observe carefully side effects of the drug. The second round of treatment was given in January 1966. The microfilaria density in 20 microliter of blood of those who received the drug was checked four times; before the treatment, during the first round of the treatment, 2 weeks and 4 months after the completion of the first round. The pre treatment mean microfilaria density of 104.6 diminished to nearly zero (only two cases with one microfilaria respectively) 2 weeks after the first round and again slightly rose up to 0.5 four months after the first round. These results indicate that DEC (Supatonin) is highly effective to eliminate the microfilaria of B. malayi. However, severe side-effects, e.g. fever (average 38.6C, maximum 39.7C), headache, backache and seldom abdominal discomfort ect. were observed. There were two cases of withdrawal from the scheme due to refusal. PMID- 12886098 TI - [The egg detection rates of Enterobius vermicularis among school children in the various regions] AB - A total of 860 cellotape anal swab specimens from 326 school children of urban area, 243 of rura1 farm village and 291 of rural fishing village was submitted for Enterobius vermicularis egg detection from September through November 1985. Each specimen was collected in the morning. Among the total 860 children's submitting specimens, 316 (36.7 per cent) were positive for E. vermicularis eggs. Of the 326 children in the urban area, E. vermicularis eggs were detected in the 45 (13.8 per cent). This detection rate was lower significantly as compared with 143 (55.6 per cent) of 243 children in the farm village or 136 (46.7 per cent) of 291 children in the fishing village. The egg detection rate in the male children was lower remarkably than that in the female children. PMID- 12886099 TI - [Intestinal parasite survey in Seoul by stool examination at Hanyang University Hospital] AB - The present study was undertaken to evaluate the present status of intestinal parasitic infection in Seoul area, Korea. During the period from June 1985 to July 1986, a total of 5,251 stool samples were collected in Department of Clinical Pathology, Hanyang University Hospital and examined by formalin-ether sedimentation technique once for helminth ova and protozoan cysts. The results were summerized as follows: The overall egg positive rate of intestinal helminthes was 2.53 per cent; and 1.43 per cent for Clonorchis sinensis, 0.7 per cent for Trichuris trichiura, 0.13 per cent for Metagonimus yokogawai, 0.06 per cent for hookworm, and 0.02 per cent for Ascaris lumbricoides, Diphyllobothrium latum and Hymenolepis nana, respectively. The overall cyst positive rate of intestinal protozoa was 1.07 per cent. Cyst positive rate was 0.06 per cent for Entamoeba histolytica and 0.13 per cent for Giardia lamblia, respectively. Higher prevalence of intestinal parasitic infection was observed in male and in 21-50 year-old groups. The highest prevalence of Clonorchis sinensis infection was observed in 40th age group(3.4 per cent), and male (2.3 per cent) was more infected than female (0.7 per cent) in general. PMID- 12886100 TI - [A survey on intestinal parasites of soldiers in Korea] AB - Total of 2,643 Korean soldiers were examined of their stool for parasitic infections by both cellophane thick smear and formalin-ether concentration techniques from August 1983 to December 1985. Out of them, 73.6% were free from any parasite, 22.6% were ova positive and 4.0% cyst positive. The ova positive rates by species were Ascaris lumbricoides 2.0%, Trichuris trichiura 13.0%, hookworm and Trichostrongylus orientalis 0.08% respectively, Clonorchis sinensis 7.6%, Metagonimus yokogawai 1.1%, Paragonimus westermani 0.08%, Echinostoma hortense 0.04%, Fibricola seoulensis 0.9%, Taenia sp. 0.3%, Hymenolepis nana 0.2% and H. diminuta 0.04%. Most of them (87.3%) were positive by a species, 11.9% by two and 0.8% by 3 species. The cyst positive rates were Entamoeba histolytica 0.6%, E. coli 1.4%, Endolimax nana 1.7%, Iodoameba butschlii 0.04% and Giardia lambla 0.9%. Among the cyst positives, 89.5% were positive by a species, 7.6% by two and 0.5% by 3 species. The intestinal parasite infections among the Korean soldiers decreased distinctly compared with previous data. PMID- 12886101 TI - Trachelobdella sinensis Blanchard, 1896 found from Cyprinus carpio nudus in Korea. AB - Trachelobdella sinensis was found in Cyprinus carpio nudus. The morphological chararacteristics were described. It is the first record of Trachelobdella sinensis in Korea(summarized from text). PMID- 12886102 TI - Larval anisakids collected from the yellow corvina in Korea. AB - Larval anisakids found in the yellow corvina (Pseudosciaena manchurica), a marine fish caught in the Yellow Sea, were classified by their morphological types. Total 1,068 anisakid larvae were collected from 30 fish examined, with the average number per fish of 35.6. They were classified into Anisakis type I larvae of Berland (859 in number, 80.4%), Contracaecum type A of Koyama et al. (13, 1.2%), Contracaecum type C'(new type) (55, 5.1%), Contracaecum type D of Koyama et al.(18, 1.7%), Contracaecum type D'(new type) (77, 7.2%), Contracaecum type V of Yamaguti(3, 0.28%), Raphidascaris sp. of Koyama et al. (1, 0.09%) and unidentified (42, 3.9%). Contracaecum type C' and D' were considered new in the literature. PMID- 12886103 TI - [Effect of flubendazole against Ascaris lumbricoides, Trichocephalus trichiurus and Enterobius vermicularis in infected children] AB - Effectiveness of flubendazole, a newly introduced as an anthelmintic in Korea, was evaluated in 28 ascariasis, 28 whipworm infections and 17 pinworm infections in primary school children. All patients were orally given by flubendazole of 500 mg once. Stools for roundworm and whipworm and anal swabs for pinworm infections were examined both before and 25 days after treatment. After flubendazole treatment, all ascariasis and pinworm infected cases were cured. On the other hand, in whipworm infections, the cure rate was 82.1 percent (23 cured of 28 infected), and the egg reduction rate was 67.1 percent, respectively. Headache (one case), dizziness (2 cases) and abdominal pain (3 cases) were noticed as untoward effects, but they were mild and disappeared within 24 hours. These results demonstrate the excellent effectiveness of flubendazole in treating these helminth infections. PMID- 12886104 TI - Epidemiological studies on malayan filariasis in an inland area in Kyungpook, Korea 3. Ecological survey of vector mosquitoes of Brugia malayi. AB - The species composition, population density, and seasonal prevalence of vector mosquitoes in an inland area of Kyungpook province were studied, based on light trap and human bait trap collections, and the recent patterns of infestation for infective larvae of Brugia malayi in these vector hosts were investigated from May to November in 1985. Nine species in four genera of mosquitoes were collected by light trap, human bait trap, and/or by nets. Anopheles sinensis Wiedemann was the most abundant species collected by light traps during this year. Culex tritaeniorhychus Giles was the second abundant species, and Aedes vexans nipponii(Theobald) ranked third in total abundance. The earlist time when A. sinensis were found was the middle of May. At that time the temperature ranged from 14.3 degrees C to 22.8 degrees C and the humidity 53-90 per cent. The month of highest average nightly catch was July, when the temperature was betweeen 21.5 degrees C and 30.6 degrees C and the humidity 72-91 per cent. The peak time of biting activity of mosquitoes was different in each month, i.e. between 22:00 23:00 in July, and 20:00-21:00 hours in September, when the temperature was between 24.3 degrees C and 26.5 degrees C and the humidity 73-88 per cent in the field. While infective larvae of B. malayi were reported to have been found in one species of mosquito in 1975, no larvae were found in any species collected in this survey. PMID- 12886105 TI - Evaluation of enzyme-linked immunosorbent assay in serological diagnosis of human neurocysticercosis using paired samples of serum and cerebrospinal fluid. AB - The applicability of micro-ELISA was evaluatd in human neuro-cysticercosis using paired samples of serum and CSF. A total of 355 cases who were mostly neurologic patients was subjected. Cystic fluid of C. cellulosae was used as antigen in protein concentration of 2.5 micro-g/ml. Serum was diluted to 1:100 and CSF was undiluted in the assay for the specific IgG antibody level. The differential criterion of the positive reaction was the abs. of 0.18 in both samples. The results were summarized as follows: The overall sensitivity of the micro-ELISA in 71 confirmed neurocysticercosis was 90.1%; the sensitivity by serum was 77.5% and that by CSF was 83.1%. CSF was a more sensitive and valuable material. Most of the false negative cases of neuro-cysticercosis showed far lower level of abs. rather than marginal. The overall specificity of the micro-ELISA in 52 confirmed other neurologic diseases was 88.5%; the specificities by serum and by CSF were 94.2% respectively. Cases of other neurologic diseases did not show false positive reactions in both samples. When serum was assayed, taeniasis(2/18), sparganosis(2/20), paragonimiasis(1/56), clonorchiasis(1/15) and fascioliasis(1/1) cases showed cross reactions. When CSF was assayed, 2 of 10 neuro-sparganosis showed cross reactions while none of 9 neuro-paragonimiasis showed it. Out of 71 confirmed neuro-cysticercosis cases, 6 and 11 showed cross reactions by serum and CSF to crude extract antigen of sparganum; but no case did show it to crude extract antigen of Paragonimus westermani. Ventricular CSF showed low or negative levels of IgG antibody than lumbar CSF unless the lesion was at the lateral ventricle itself. Out of 4 racemose cysticercosis cases, 3 showed positive reaction in serum while all of 3 examined CSF were positive. The above results indicated that the serological test for detecting the specific IgG antibody by micro-ELISA using paired samples of serum and CSF was very helpful for clinical differentiation of neuro-cysticercosis from neurologic diseases of other causes. PMID- 12886106 TI - [The karyotype of Paramphistomum explanatum (Creplin, 1849) obtained from Korean cattle] AB - As a series of systematic classification of paramphistomes, in the first step, paramphistomes in the rumen and reticulum were collected on 170 Korean cattles (2 3 years age, male) slaughtered at Jeonju abattoir from July 1984 to September 1985 and were classified by means of morphology of the worms. Afterwards, the karyotype of Paramphistomum explanatum (Creplin, 1849) which is the common in Korean cattle was detected by means of modified air-drying method from testis cells of the worm. The following is a brief summary of the leading facts gained through the experiment. Most of the cattle slaughtered at the abattoir were infected with paramphistomes. The 5 species of the worms were detected on 170 Korean cattle and the worm burden per head was from 2 to 784 (on the average 170) worms, 120(70.59 percent) heads out of them involving 2-100 worms. In 28,900 individuals of paramphistomes obtained on 170 Korean cattle, appearance rates of various worms were as follows : 49.74 percent in P. explanatum, 48.08 percent in P. cervi, 0.98 percent in Orthocoelium orthocoelium, 0.89 percent in Fischoederius cobboldi and 0.14 percent in Cotylophoron cotylophorum. The chromosome number of 620 P. explanatum in the haploid and diploid cells was n=9 and 2n=18, and abundant cells in meiotic division were observed; 1,420 haploid and 38 diploid cells were reliable. Nine pairs of mitotic chromosomes were homologous and the chromosomes were composed of five medium-sized metacentrics (m), subtelocentrics (st) or submetacentrics (sm)and four small-sized subtelocentrics (st) or submetacentrics (sm), while meiotic metaphase chromosomes were composed of five medium and four small-sized. The haploid of the testis cells showed C-band in the centromeric region from 8 of them, whereas the remaining chromosome No. 5 included heterochromatin on the tip region, and chromosomes No. 3 and No. 7 showed a remarkable C-band distinguished from other chromosomes. PMID- 12886107 TI - [Antigenicity of the soluble egg antigen of Paragonimus westermani] AB - To evaluate the immature eggs of Paragonimus westermani as a source of diagnostic antigen, about a million eggs which were excreted by 104 adult worms were collected; their saline extract (soluble egg antigen; PwSEA) was prepared. The specific IgG and IgM antibody levels were observed in experimental dog paragonimiasis by micro-ELISA, using PwSEA as well as whole worm extract of 12 week-old P. westermani (PwWWE). The protein composition of the PwSEA was observed by disc-PAGE. The results could be summarized as follows: Specific IgG antibody to PwSEA began to increase on 8 weeks after the experimental infection; it maintained its high level until the observation period of 13 weeks. The levels of IgM antibody to PwSEA, however, did not show any significant change. Specific IgG antibody to PwWWE began to increase earlier from 2 weeks after the infection and continued to increase until the observation period of 13 weeks. Its level was much higher than that to PwSEA. Specific IgM antibody to PwWWE increased temporarily during 2-8 weeks after the infection. By disc-PAGE, PwSEA showed 2 protein bands of very low motility. The bands of PwSEA corresponded to the first and second bands in the electrophoretic pattern of PwWWE of the 12 week-old worms. The above results indicated that the PwSEA induced antibody production in dog paragonimiasis but its antigenicity was weaker than PwWWE to be used as a diagnostic antigen. PMID- 12886108 TI - Growth and development of Pygidiopsis summa in rats and mice with a supplementary note on its morphological characters. AB - The growth and development of Pygidiopsis summa were studied in experimental rats and mice, and a special reference was given to its morphological characters differed from the type species, P. genata. The metacercariae were obtained from young mullets (Mugil sp.), and total 21 rats and mice infected each with 1,000 metacercariae. Worms of various ages of infection, from 3 to 28 days, were subjected to a microscopic observation. The worms grew rapidly and remarkably in size up to 10 days of infection, to become 0.53-0.82 mm long and 0.31-0.39 mm wide, but nearly stopped the growth thereafter. Their genital organs developed more rapidly and fully matured within 3-5 days. At 3 days several eggs were found in uterus. The presence of two groups of small spines, 5-6 on the right and 7-9 on the left side of the genital apparatus, was a new finding in this study and considered a distinct character of P. summa. The morphology of ventral sucker and intestinal ceca was also different from P. genata. This study confirms the validity of the species, P. summa. PMID- 12886109 TI - [Tegumental ultrastructures of Echinostoma hortense observed by scanning electron microscopy] AB - The tegumental ultrastructures of Echinostoma hortense adults were observed by scanning electron microscopy. The worms of 4 weeks of age were harvested from albino rats experimentally infected with the metacercariae obtained from the loach. The results were as follows: The worms were leaf-like and their anterior end portion, including oral sucker and head crown, ventrally curved to face posteriorly. The tegument of whole body was wrinkled transversely and covered with cobblestone-like cytoplasmic processes. The oral sucker had roundly swollen (type II) sensory papillae on the ventral half of its lip and uni-ciliated knob like (type I) sensory papillae, arranged in 2-3 rows, on the dorsal outer surface. Aspinous ventral sucker had many of type I papillae arranged in a circular band on its outer surface. The tegument around the genital opening was of similar feature to the ventral sucker, but sensory papillae were hardly found around the former. Scale-like spines with broad base and round tip were distributed densely on the tegument anterior to the ventral sucker but they became sparse in posterior half of the ventral surface, finally to disappear at posterior extremity. A few number of type I papillae were observed on the ventral surface. The results suggest that the tegument of E. hortense is similar to that of other echinostomes especially E. revolutum. But the number and arrangement of collar spines, and/or the type and distribution of sensory papillae seem characteristic features of E. hortense differed from other echinostomes. PMID- 12886110 TI - [Two cases of human infection by Echinostoma cinetorchis] AB - Eggs of Echinostoma sp. were found from two patients who were hospitalized between Oct. and Dec. 1985. Single dose of 10 mg/kg of praziquantel was prescribed to each patient and after purgation with magnesium salt seven and two adult worms of Echinostoma cinetorchis in respect were recovered from patients. The first case was a 54-year-old male residing at Chungju city in Chungbuk province. He was hospitalized because of severe weight loss and frequent urination. The second case was a 71-year-old male residing at Chungsun-kun in Kangwon province, and hospitalized due to suspected lung cancer. The average size of the eggs found in the stools was 98 x 63 micrometer (the first case) and 100 x 67 micrometer (the second case) respectively, and total nine worms collected were identified as E. cinetorchis. PMID- 12886111 TI - [Two cases of natural human infection by Echinostoma hortense] AB - Two cases of human Echinostoma hortense infection were found in Seoul Paik Hospital. Their stools revealed echinostomatid eggs. After treatment with praziquantel (single does of 10-12 mg/kg) and purgation with magnesium salt, total 21 flukes were collected in one case. The flukes were 5.9-7.5 mm long, had 27-28 collar spines around their head, laterally deviated ovary and two tandem testes. They were identified as E. hortense Asada, 1926. The cases are 38-year and 20-year old men residing in Seoul, whose hometown is Chungsong-gun and Seungju-gun, Kyongsangbuk-do respectively. They had eaten raw flesh of various kinds of fresh water fishes (both cases) and/or salamander (latter case), which are considered possible sources of this fluke infection. They experienced abdominal discomfort and/or diarrhea. Hematology revealed 22 percent eosinophilia in the latter case, but it became 5 percent after the treatment. Echinostomatid eggs were not found after the treatment in both cases. These are the 4th and 5th human cases of E. hortense infection in Korea. PMID- 12886112 TI - Human infections by Heterophyes heterophyes and H. dispar imported from Saudi Arabia. AB - Two human cases of Heterophyes heterophyes and H. dispar infections were proven by the recovery of their adult worms. The cases were 38-year and 40-year old Korean workers who had been in Saudi Arabia for 4-6 years and returned home in 1983 or 1984 with gastrointestinal troubles. In Saudi Arabia they had eaten raw brackish water fishes such as the mullet. After the treatment with 10 mg/kg praziquantel and purgation with magnesium salt, a total of 19 specimens of H.heterophyes and 140 of H. dispar were collected. It is of interest that the worms persisted in a patient although he had been back in Korea for 14 months. This is the first report on imported heterophyiasis in Korea. Human infection by H. dispar is the first record in the literature. PMID- 12886113 TI - A human case of gallbladder fascioliasis in Korea. AB - The authors would record the fifth case of human fascioliasis in Korea, occurring in a 48 year old Korean man who lived in Munkyong, Kyongsangbuk-do. The diagnosis was based on the morphology and measurements of the eggs which were collected in necrotic debris from the tunnel-like ulcerations of the gallbladder wall which was surgically removed for a gall stone. Although we failed to find out the fluke in the specimen, there found an evidence of once presence of Fasciola in the inflamed wall of the gallbladder in this patient. PMID- 12886114 TI - Ten human cases of Fibricola seoulensis infection and mixed one with Stellantchasmus and Metagonimus. AB - Ten soldiers of the Korean Army were found to have F. seoulensis ova in their faeces. These men were treated with praziquantel and the worms passed after purging were successfully collected in one case: 59 F. seoulensis, 68 Metagonimus and 2 Stellantchasmus falcatus were collected. 2 men were also infected with Clonorchis sinensis. All the cases of fibricoliasis had eaten raw or undercooked snake meat or frogs during survival training. All were egg-negative 2-6 months after treatment. PMID- 12886115 TI - Family caregivers' sleep loss and depression over time. AB - Depression is a normal response when a family member receives a diagnosis of cancer. However, this response may be exacerbated by other factors such as chronic sleep loss that are amenable to intervention. This pilot study described caregiver sleep and depression patterns over time and explored the feasibility of data collection methods and instruments. The stress and coping framework of Lazarus and Folkman guided this study. A descriptive correlational design was used for this 10-week pilot study. Ten adult family caregivers of patients with cancer were recruited from outpatient oncology clinics. Sleep quality and depression were measured weekly. Actigraphs were worn for 72 hours during weeks 1, 5, and 10. Individual sleep quality and depression scores were generated. Actigraph latency, duration, and efficiency scores were generated. Actigraph and sleep quality scores were compared. Individual caregiver sleep and depression plots show large variance over time. Discrepancies were noted between Actigraph and sleep quality latency, duration, and efficiency scores. Sleep and depressive symptoms fluctuate widely over time. Therefore, accurate assessment and treatment of caregiver problems require repeated assessments. Self-reports of sleep and depression appear to underestimate problems and must be evaluated carefully within this context. PMID- 12886116 TI - Spiritual needs of patients with cancer and family caregivers. AB - The purpose of this descriptive, cross-sectional, qualitative study was to describe the spiritual needs experienced in living with cancer from the perspective of patients with cancer and family caregivers. The sample included 28 African American and Euro-American patients with cancer and family caregivers receiving care from inpatient and outpatient units at two metropolitan hospitals in the southwestern United States. In-depth, tape-recorded, semistructured interviews were analyzed using the process of data reduction, data display, and verification. Seven categories of identified spiritual needs included needs associated with relating to an Ultimate Other; the need for positivity, hope, and gratitude; the need to give and receive love; the need to review beliefs, the need to have meaning; and needs related to religiosity and preparation for death. Informants responded with varying levels of awareness of personal spiritual needs. Caregivers were observed to have spiritual needs similar to those of patients. The findings of this study will inform nurses as they assess and document spiritual needs. PMID- 12886117 TI - Family experience caring for terminally ill patients with cancer in Hong Kong. AB - In this study, 24 family caregivers of terminally ill patients participated in in depth interviews regarding their experiences of giving care. The data were analyzed using grounded theory qualitative method. Commitment emerged as the precondition of the caregiving process. The caregivers did not perceive the work of caring as a burden. Rather, they felt that despite any personal hardships, what they were doing was important to their loved ones and therefore meaningful to them as caregivers. The components of commitment can be described as relational commitment, the act of showing love, and determination. The process of caregiving includes four phases: 1) holding onto hope for a miracle, 2) taking care, 3) preparing for death, and 4) adjusting to another phase of life. A patient-caregiver relationship, Confucian concepts of yi (appropriateness or rightness), and filial duty are reflected in the process of caregiving. Consequences of the process include finding meaning in life and peace of heart and mind. The emotional aspect of the caregiving experience can be described as an intense emotional experience filled with feelings of hope and hopelessness, guilt, fear, and regret. As a result of the caregiving experience, most participants found they have had a change of worldviews and treasure their lives. Findings show a significant need for interventions at each phase of the caregiving process designed to provide effective and culturally sensitive support and affirmation to family members as they care for their loved ones with terminal illnesses. PMID- 12886118 TI - The difficulties faced by informal caregivers of patients with terminal cancer in Hong Kong and the available social support. AB - This study aimed to explore the difficulties experienced by primary informal caregivers of Chinese patients with terminal cancer in Hong Kong and the social support available for such caregivers. For this exploratory study, 21 primary informal caregivers of patients with terminal cancer were recruited from a hospice home service to complete a questionnaire. The results show that all but one of the caregivers (95.2%) perceived difficulties in rendering care. They reported experiencing four major kinds of difficulties: relationship with the care receiver (n = 11, 52.4%), emotional reactions to caring (n = 9, 42.9%), physical demands (n = 10, 47.6%), and restricted social life (n = 11, 52.4%). Of the 21 caregivers, 20 (95.2%) felt that the support received from hospice home care nurses was useful, especially in skill training (n = 16, 76.2%), informational support (n = 20, 95.2%), and emotional support (n = 20, 95.2%). Informal caregivers also indicated that home care nurses provided more useful overall support than family members and friends (chi2 = 11.35; P =.003). With a better understanding of the difficulties experienced by caregivers and the support they receive, hospice home care nurses will be in a better position to identify effective strategies for helping informal caregivers cope with their difficult circumstances. PMID- 12886119 TI - An environmental intervention to restore attention in women with newly diagnosed breast cancer. AB - Earlier research indicated that attentional fatigue with reduced capacity to direct attention in women treated for breast cancer may be ameliorated by a theoretically based intervention involving regular exposure to the natural environment. This study tested the efficacy of a natural environment intervention aimed at restoring attention in 157 women with newly diagnosed breast cancer. Capacity to direct attention was assessed with a brief battery of objective measures at two time points: approximately 17 days before surgery (time 1) and 19 days after surgery (time 2). A randomly assigned intervention protocol was initiated after the first assessment and before any treatment. The intervention comprised a home-based program involving 120 minutes of exposure to the natural environment per week. The intervention group (n = 83) showed greater recovery of capacity to direct attention from the pretreatment (time 1) to the preadjuvant therapy period (time 2), as compared with the nonintervention group (n = 74). A significant effect of the natural environment intervention was observed even after control was used for the effects of age, education, attention scores at time 1, other health problems, symptom distress, and extent of surgery. The findings suggest therapeutic benefits for capacity to direct attention from early intervention aimed at restoring attention in women with newly diagnosed breast cancer. PMID- 12886120 TI - Attainment and importance of life values among patients with primary breast cancer. AB - The main aims of this study were to investigate the extent to which women with recently diagnosed primary breast cancer (N = 29) and matched control subjects without cancer (N = 29) differ in perceived attainment and importance of life values and to study prospectively life value ratings during 1 year in a large group of recent attendees at mammography screening (N = 706). Life values were assessed by a study-specific version of a life value questionnaire, including ratings of the perceived attainment and importance of seven life value dimensions. Women with a recent diagnosis of primary breast cancer were found to attribute significantly more importance to positive relations than healthy controls. No other differences between these groups were found regarding the attainment or importance of life values. Perceptions of life values were found to vary as a function of age, marital or cohabitation status, and parenthood, and to be stable over a 9-month period in screening attendees. The implications of the current findings for the understanding of women's psychological adaptation to breast cancer are discussed. PMID- 12886121 TI - Quality of life among primary caregivers of Taiwanese children with brain tumor. AB - This study evaluated the quality of life among primary caregivers of Taiwanese children with brain tumors and examined the factors associated with their quality of life. Data were obtained from the Impact of Childhood Cancer on the Chinese Family survey conducted by the Taiwan Childhood Cancer Foundation in 2001. This analysis included 30 families. Quality of life among primary caregivers was measured by the World Health Organization Questionnaire on Quality of Life: BREF Taiwan version. Quality-of-life scores for various subgroups were compared using the Mann-Whitney U test or the Kruskal-Wallis test. The scores in the physical health and psychological domains for caregivers of children with brain tumors were lower than the norm scores for healthy adults. Caregivers of boys had higher scores in social relationships. Caregivers of children within 5 years of the brain tumor diagnosis had higher scores in the physical health domain. The child's treatment stage was associated with caregiver scores in the physical health and psychological domains. The caregiver's psychological domain was negatively affected by a low education level. The health status of caregivers was positively associated with their quality of life in physical health, psychological status, social relationships, and environment domains. The lower quality of life among caregivers indicates the need for promoting an awareness in healthcare providers of the impact that having a child with brain tumor has on the quality of life. Family-centered interventions are implicated. Healthcare providers should elicit information regarding the caregiver's perceived physical health and intervene early. This may help to improve the quality of life among caregivers of children with brain tumors. PMID- 12886122 TI - Quality of life immediately after peripheral blood stem cell transplantation. AB - This prospective, longitudinal study investigated the quality of life among patients immediately after undergoing peripheral blood stem cell transplantation. The patients were assessed before transplantation (n = 16), immediately before hospital discharge (n = 10), 2 weeks after hospital discharge (n = 10), and 6 weeks after hospital discharge (n = 8). Patients completed the European Organization for Research and Training Quality of Life Questionnaire-Core 30 and the Ferrans and Powers Quality of Life Index at each assessment point. A brief telephone interview was conducted 6 weeks after hospitalization (n = 7). The study results indicate that patients undergoing peripheral blood stem cell transplantation experience significant appetite loss (P <.01), diarrhea (P <.05), nausea and vomiting (P <.05), and sleep disturbances (P <.01) during the acute posttransplantation period. Other clinically important changes also were identified, so that a general pattern of recovery emerged. These patients reported diminished functional ability, increased symptoms, and poorer quality of life immediately before hospital discharge, with a return to pretransplantation levels by 6 weeks after hospitalization. In contrast, life satisfaction continued to decline 2 weeks after hospitalization, but did improve by 6 weeks. The qualitative data provide further support for this trend. PMID- 12886123 TI - Prescriptions for health providers: from cancer patients. AB - Women enter an increasingly complex medical system when they receive the diagnosis of breast cancer. Subsequently, they are confronted with numerous complicated, confusing, and less than certain decision-making episodes. Research was undertaken to follow and observe in vivo medical encounters between women and their oncologic physicians throughout the breast cancer diagnostic and treatment process. Empirical data was collected using qualitative techniques to deepen understanding of the decision-making process in the oncology setting. Physicians and patients alike make assumptions about each other that can hinder the development of a trusting and caring attitude that enables a more efficient delivery of healthcare. Patients offer recommendations for care to their physicians. In addition, ethical concerns regarding how medical options are presented to patients emerged in this study. PMID- 12886124 TI - Tropisetron and dexamethasone administered twice daily for the prevention of acute emesis in patients treated with continuous infusion of Cisplatin Doxorubicin and high-dose Ifosfamide over 48, 24, and 120 hours. AB - The antiemetic effectiveness of 5-HT3 receptor antagonists in combination with dexamethasone in patients receiving short-term infusion chemotherapy has been well demonstrated. Less information is available about the efficacy of the same antiemetic combination in patients treated with regimens of chemotherapy in which the drugs are delivered in continuous infusion of several hours. The purpose of this study was to report the effectiveness of a double administration of antiemetic drugs in patients treated with strong emesis-inducing drugs for several days. In this study, 19 male and 13 female patients with osteosarcoma, ages 9 to 45 years, treated with chemotherapy, received intravenous tropisetron 5 mg plus dexamethasone 8 mg every 12 hours during the first two cycles of the preoperative treatment: cisplatin 120 mg/m2 over 48 hours followed by Adriamycin 75 mg/m2 delivered in 24 hours and continuous infusion of ifosfamide 15 g/m2 over 120 hours. The assessment of the antiemetic efficacy was performed three times every day: from 8:00 am to 4:00 pm, from 4:00 pm to 12:00 am, and from 12:00 am to 8:00 am. The patients were followed from the beginning of the treatment until 2 hours after its end, when they were discharged from hospital. Complete protection from emesis was obtained in 80% of the 256 days of treatment: 81% during the first cycle (cisplatin 120 mg/m2 in 48 hours followed by Adriamycin 75 mg/m2 delivered in 24 hours) and 79% during the second cycle (continuous infusion of ifosfamide 15 g/m2 in 120 hours). In both cycles, complete protection declined from the first to the last day of treatment (from 100% to 62% during the first cycle and from 100% to 63% during the second cycle). These results indicate that when chemotherapy is administered in a protracted infusion, higher doses of antiemetic agents are necessary to achieve acceptable antiemetic activity. PMID- 12886125 TI - Infectious pulmonary nodules in immunocompromised patients: usefulness of computed tomography in predicting their etiology. AB - PURPOSE: To review the high-resolution computed tomography (CT) findings in immunocompromised patients who had nodular opacities and a proven diagnosis to determine whether the various infectious pulmonary nodules have distinguishing features on CT. MATERIALS AND METHODS: The high-resolution CT scans obtained in 78 immunocompromised patients with solitary or multiple nodular opacities of proven infectious etiology were reviewed retrospectively by 2 independent thoracic radiologists. Patients whose predominant abnormality consisted of branching linear or nodular opacities (tree-in-bud pattern) characteristic of infectious bronchiolitis and endobronchial spread of tuberculosis were excluded. The CT scans were assessed for the presence, appearance, size, and distribution of parenchymal nodules. Relations between findings at CT and the different infectious etiologies of nodules were assessed with regression analysis. Agreement between the 2 observers was assessed using the kappa statistic. RESULTS: The infectious causes included mycobacteria (n = 24), fungi (n = 22), bacteria (n = 20), and viruses (n = 12). Multivariate analysis demonstrated that a diameter <10 mm was the only independent predictor of etiology (P < 0.0001) and that patients whose nodules all measured less than 10 mm in diameter were most likely to have a viral infection. Nodules limited in size to less than 10 mm in diameter were seen in 83% of viral infections compared with 5% of bacterial infections (odds ratio [OR] = 95.0; 95% confidence interval (CI): 6.08-4,321.5, P < 0.0001), 0% of mycobacterial infections (OR = 91.7; 95% CI: 7.21-4,090.22, P < 0.0001), and 14% of fungal infections (OR = 31.67; 95% CI: 3.56-375.09, P = 0.0003). CONCLUSION: Although some overlap exists, nodule size is helpful in the differential diagnosis of infectious causes of nodules in immunocompromised patients. Patients whose nodules are all less than 10 mm in diameter are most likely to have a viral infection. PMID- 12886126 TI - Solitary hot spots in the ribs on bone scan: value of thin-section reformatted computed tomography to exclude radiography-negative fractures. AB - PURPOSE: The purpose of this study was to classify solitary, scintigraphy positive and radiography-negative rib lesions and to clarify the features of rib fractures by using thin-section reformatted helical computed tomography (CT). METHODS: Forty-seven patients whose whole-body bone scans showed a solitary hot spot in a rib as their first abnormal scintigraphic finding and whose plain radiograph showed negative or indeterminate results were examined by thin-section reformatted CT. Final diagnosis was based on follow-up over a 20-month period. RESULTS: The final diagnosis included 17 cases of fractures where CT findings were fracture line, focal sclerosis, and callus formation. Fourteen ribs demonstrated intramedullary, focal osteosclerosis, and 8 ribs did not demonstrate any abnormalities. Four metastatic lesions appeared as intramedullary mixture of osteolysis and osteosclerosis, or bone destruction. Four intramedullary lesions with cystic appearance remained unchanged. CONCLUSIONS: Thin-section reformatted helical CT delineated minute, radiographically occult fractures of the rib. Excluding fractures, helical CT can avoid further unnecessary examinations. PMID- 12886127 TI - Lung cysts in subacute hypersensitivity pneumonitis. AB - PURPOSE: To determine the frequency of cystic lesions on high-resolution computed tomography (CT) in patients with subacute hypersensitivity pneumonitis. METHODS: High-resolution CT scans in 182 patients with proven subacute hypersensitivity pneumonitis were retrospectively evaluated for the presence of lung cysts. Patients with CT evidence of emphysema or interstitial fibrosis and patients with connective tissue disease were excluded from the study. Two thoracic radiologists reviewed the thoracic CT scans for the presence and location of lung cysts. RESULTS: Thin-walled lung cysts were identified in 24 (13%) of 182 patients and ranged in size from 3 to 25 mm in maximal diameter. The patients included 16 men and 8 women (age range, 30-79 years; mean age, 48 years). The cysts ranged from 1 to 15 in number (mean, 4 cysts) and had a random distribution. Sixteen of the 24 patients also had areas of decreased attenuation and vascularity consistent with air trapping. CONCLUSIONS: Thin-walled cysts can be seen in a small percentage of patients with subacute hypersensitivity pneumonitis. The cysts resemble those seen in lymphocytic interstitial pneumonia, and their pathogenesis is uncertain. PMID- 12886128 TI - Improved radiologic staging of lung cancer with 2-[18F]-fluoro-2-deoxy-D-glucose positron emission tomography and computed tomography registration. AB - PURPOSE: To determine if volumetric nonlinear registration or registration of thoracic computed tomography (CT) and 2-[18F]-fluoro-2-deoxy-D-glucose-positron emission tomography (FDG-PET) datasets changes the detection of mediastinal and hilar nodal disease in patients undergoing staging for lung cancer and if it has any impact on radiologic lung cancer staging. METHOD: Computer-based image registration was performed on 45 clinical thoracic helical CT and FDG-PET scans of patients with lung cancer who were staged by mediastinoscopy and/or thoracotomy. Thoracic CT, FDG-PET, and registration datasets were each interpreted by 2 readers for the presence of metastatic nodal disease and were staged independently of each other. Results were compared with surgical pathologic findings. RESULTS: One hundred and thirty lymph node stations in the mediastinum and hila were evaluated each on CT, PET, and registration datasets. Sensitivity, specificity, positive predictive value, and negative predictive value, respectively, for detecting metastatic nodal disease for CT were 74%, 78%, 55%, 88%; for PET with CT side by side, 59% to 76%, 77% to 89%, 48% to 68%, and 84% to 91%; and for CT-PET registration, 71% to 76%, 89% to 96%, 70% to 86%, and 90% to 91%. Registration images were significantly more sensitive in detecting nodal disease over PET for 1 reader (P = 0.0156) and were more specific than PET (P = 0.0107 and 0.0017) in identifying the absence of mediastinal disease for both readers. Registration was significantly more accurate for staging when compared with PET for both readers (P = 0.002 and 0.035). CONCLUSION: Registration of CT and FDG-PET datasets significantly improved the specificity of detecting metastatic disease. In addition, registration improved the radiologic staging of lung cancer patients when compared with CT or FDG-PET alone. PMID- 12886129 TI - Location, size, and distribution of mediastinal lymph node enlargement in chronic congestive heart failure. AB - PURPOSE: The purpose of this study is to identify the prevalence, location, and size of enlarged mediastinal lymph nodes in patients with chronic congestive heart failure and to correlate the presence of lymph node enlargement with cardiac ejection fraction. METHODS: Sixty-six consecutive, retrospectively identified patients underwent computer tomography (CT) imaging of the thorax as part of a routine work-up prior to cardiac transplantation from 1993 to 1996. CT images of 44 of these patients were independently examined by 3 radiologists for evidence of pulmonary edema, pleural effusions, and the presence, size, and location of lymph nodes >1 cm in short axis. Multigated acquisition (MUGA) scans were available for cardiac ejection fraction assessment in 38 of the 44 patients. RESULTS: Twenty-nine (66%) patients had at least 1 mediastinal lymph node >1 cm. The mean ejection fraction was significantly less for patients with lymph node enlargement when compared with patients without lymph node enlargement (20% versus 35%; P < 0.01). Adenopathy was observed in 81% of patients with a calculated ejection fraction of <35%. No patient with an ejection fraction of >35% had lymph node enlargement. There was no correlation between pulmonary edema and the frequency of lymph node appearance. Sixty-three percent of the enlarged nodes were pretracheal, with a mean short axis diameter for all the enlarged nodes of 1.3 cm. CONCLUSIONS: Enlarged mediastinal lymph nodes were observed in 81% of patients with a calculated ejection fraction of <35%, most commonly in the pretracheal group. The presence of the lymph nodes did not correlate with CT evidence of pulmonary edema. PMID- 12886130 TI - Additional coronal images using low-milliamperage multidetector-row computed tomography: effectiveness in the diagnosis of bronchiectasis. AB - PURPOSE: The aim of our study was to evaluate the effectiveness of additional coronal images using low-milliamperage multidetector-row computed tomography (CT) in the diagnosis of bronchiectasis. METHODS: Helical volumetric CT scans (120 kVp, 70 mA, 2.5-mm collimation, table speed of 15 mm/s, table rotation time of 1 second) using low-milliamperage multidetector-row CT were obtained through the thorax in 110 patients who were suspected of bronchiectasis. Both axial (2.5-mm section thickness) and coronal (1.3-2.0-mm section thickness) reformatted images were made and sent to picture archiving and communication system (PACS) monitors. Two independent observers assessed CT scans twice; with axial images only and with both axial and coronal images. The detection rates of bronchiectasis were compared between readings with axial images only and with both axial and coronal images by using a nonparametric method of clustered data. Confidence grades were given to the distribution and type of bronchiectasis. RESULTS: With axial images only, the detection rates of bronchiectasis on a per-patient basis were 97% (213/220 patients, kappa = 0.888) whereas with both axial and coronal images, the detection rates were 100% (220/220 patients, kappa = 1.000) (P = 0.0001). Confidence to the distribution of bronchiectasis was greater with both axial and coronal images than with axial images only (P = 0.008). CONCLUSIONS: Additional coronal images using low-milliamperage multidetector-row CT are effective in the diagnosis of bronchiectasis by providing enhanced detection rates and confidence to the distribution of lesions. PMID- 12886131 TI - Ground-glass computed tomography pattern in chronic beryllium disease: pathologic substratum and evolution. AB - Five cases of chronic beryllium disease with predominant or isolated diffuse ground-glass lung opacities on computed tomography are reported with correlation to lung pathology. The ground-glass attenuation was either homogeneous and isolated (n = 1) or patchy and clearly predominant (n = 4) over linear and nodular opacities. In 4 cases, histologic samples showed a prominent diffuse involvement of the alveolar-capillary walls by florid granulomas. Evolution under corticosteroid was remarkable by the appearance of small-sized cysts in 3 cases. PMID- 12886132 TI - A case of inverted left atrial appendage mimicking a tumor. AB - We report a rare case of an inverted left atrial appendage without prior cardiac surgery. A left atrial mass was incidentally found during routine echocardiography in a 19-year-old man with mitral valve prolapse. Echocardiography revealed a hyperechoic mass in the left atrium, and a neoplastic lesion could not be excluded. On magnetic resonance imaging (MRI), this mass consisted of fat tissue that showed continuation to epicardial fat, indicating an inverted left atrial appendage mimicking a tumor in the left atrium. When a mass in the left atrium is observed on echocardiography, there are several differential diagnoses, including thrombus, vegetation, and intra-atrial neoplasms such as myxomas. Recently, several studies reported cases with inverted left atrial appendages mimicking tumors in patients after cardiac operations. We present a case of inverted left atrial appendage without any prior cardiac surgery. Cardiac MRI was highly useful to obtain the final diagnosis of inverted left atrial appendage. PMID- 12886133 TI - Clinical significance of cavernous carotid calcifications encountered on head computed tomography scans performed on patients seen in the emergency department. AB - PURPOSE: To determine whether the presence of calcification in the cavernous segment of the internal carotid artery seen on head computed tomography (CT) performed in the emergency department shows any relationship to the existence of systemic disease. Significant and consistent relationships could be used as predictors in suggesting the presence of systemic disease in those patients receiving head CTs for unrelated symptoms. METHODS: A retrospective investigation was performed on 295 consecutive patients presenting to the Massachusetts General Hospital emergency department for head CT scan during the course of 1 month. Head CT images were analyzed for carotid siphon calcifications. Data regarding multiple predictors of disease including gender, age, hypertension, diabetes, smoking, hypercholesterolemia, cardiac disease, and alcohol and intravenous drug abuse were abstracted from the medical record for each patient. RESULTS: Strong univariate correlation was noted in the comparison between the presence of carotid siphon calcification and gender, age, diabetes, hypertension, smoking, and cardiac disease while alcohol and intravenous drug abuse correlated poorly. Multivariate models including categorized age and gender demonstrated a consistently strong interaction with diabetes, cardiac disease, and smoking while hypertension and hypercholesterolemia showed a loss of significance. Interaction term analysis indicated a strong (ie, significant) interaction between gender and both hypertension and hypercholesterolemia. CONCLUSIONS: Carotid siphon calcifications are strong predictors of existing medical disease in simple cases. In patients with a combination of diseases, a strong interaction between covariates is noted, suggesting a more complex interaction contributing to the formation of carotid siphon disease. PMID- 12886134 TI - Orbital lymphoma and subacute or chronic inflammatory pseudotumor: differentiation with two-phase helical computed tomography. AB - OBJECTIVE: The differentiation between orbital lymphoma and subacute or chronic inflammatory pseudotumor (SCIPT) may be difficult clinically and radiologically. The aim of this study was to evaluate the enhancement characteristics of orbital lymphoma and SCIPT with two-phase helical computed tomography (CT) and delayed coronal CT and to determine whether attenuation measurements on CT can be used to differentiate between the 2 diseases. METHODS: Nineteen histopathologically proven orbital lymphomas and 9 SCIPTs were examined with two-phase helical CT. After injecting 90 mL contrast material at a rate of 3 mL/s, early- and late phase axial CT scans were obtained with scanning delays of 30 and 90 seconds, respectively. Delayed coronal scans were obtained with delays of 4-9 minutes. Attenuation of the lesions at each phase was measured quantitatively. Relative percentages of contrast enhancement (CE) were calculated to determine the cutoff value for differentiating SCIPT from lymphoma. RESULTS: The CT attenuation change over time was significantly different between orbital lymphoma and SCIPT (P < 0.05). Increased CT attenuation between early- and late-phase axial scans was seen in 42% (n = 8) of lymphoma cases, and decreased CT attenuation was seen in 58% (n = 11). In 17 lymphomas (90%), the CT attenuation decreased between late phase axial and delayed coronal scans. Conversely, in 7 SCIPTs (78%), the CT attenuation increased gradually over time from early-phase axial to delayed coronal scans. The relative percentage of CE at the delayed coronal scan had a cutoff value of -6.97%, a sensitivity of 84%, and a specificity of 100%. CONCLUSION: Different characteristics of attenuation change on two-phase helical CT and delayed coronal CT can be helpful in differentiating between orbital lymphoma and SCIPT. PMID- 12886135 TI - Clinical and imaging findings of lymphoma in patients with Sjogren syndrome. AB - OBJECTIVE: To describe and correlate the clinical and imaging findings of lymphomas in patients with Sjogren syndrome. METHODS: The authors reviewed the medical and imaging records of 27 cases of lymphoma from among a total of 463 patients with Sjogren syndrome. The estimated prevalence of lymphoma in patients with Sjogren syndrome was 5.8%. There were 22 women and 5 men. Histopathologically, 26 of the 27 neoplasms were non-Hodgkin lymphoma, including 6 mucosa-associated lymphoid tissue lymphomas, and the other neoplasm was Hodgkin lymphoma. The clinical and imaging findings of lymphomas were analyzed. RESULTS: No obvious correlations were present between the duration or severity of Sjogren syndrome and the lymphoma development. At the initial diagnosis, extranodal involvement was observed in 14 (52%) of the 27 patients, including the salivary gland (n = 9), lacrimal gland (n = 2), lung (n = 2), and thyroid gland (n = 1), mostly in the neck organs. On the other hand, nodal involvement was observed in 21 (78%) of the 27 patients. Of these 21 patients, 19 had at least cervical lymph node involvement. CONCLUSION: Patients with Sjogren syndrome are at increased risk of lymphoma development. Because most lymphomas initially involve the neck organs, including the lymph nodes, meticulous imaging studies mainly focused on the cervical regions are recommended in the follow-up of patients with Sjogren syndrome. PMID- 12886136 TI - Usefulness of semiquantitative FDG-PET in the prediction of brain tumor treatment response to gamma knife radiosurgery. AB - PURPOSE: The aim of this study is to evaluate whether posttreatment FDG-PET can predict the treatment response to gamma knife radiosurgery (GKRS) for patients with gliomas. METHOD: Eighteen patients with histologically confirmed gliomas treated by GKRS and followed for more than 24 months were enrolled in this study. All patients underwent FDG-PET 6-8 months after GKRS. PET images and MR images were integrated and tumor-to-cortex (T/C) and tumor-to-white matter (T/WM) FDG uptake ratios were measured. By estimating the change of the maximum tumor diameter measured on the contrast-enhanced MRI (the last follow-up MRI vs. the pre-GKRS MRI), the tumor response was classified as complete remission, partial remission (PR), stable disease (SD), and progression of disease (PD). Group 1 includes four tumors with PR and 6 tumors with SD; group 2 includes 8 tumors with PD after GKRS. RESULTS: Both mean T/C ratio and mean T/WM ratio of group 1 patients are statistically less than those of group 2 patients. CONCLUSION: Our study suggests that, for patients with gliomas treated by GKRS, the posttreatment FDG-PET may be useful to predict the treatment response. PMID- 12886139 TI - Nonketotic hyperglycinemia: diffusion magnetic resonance imaging findings. AB - We report about a boy with nonketotic hyperglycinemia who was studied at 15 days of life with a follow-up examination at age 6 months. Magnetic resonance (MR) imaging revealed progressive atrophy, callosal thinning, and delayed myelination. Glycine peaks were shown by proton MR spectroscopy at 3.56 ppm with a long echo time (TE, 135 milliseconds; TR, 1500 milliseconds). Echo-planar diffusion MR imaging (TR, 5700 milliseconds; TE, 139 milliseconds) at 15 days of life revealed high-signal lesions in the pyramidal tracts, middle cerebellar pedicles, and dentate nuclei on b = 1000 s/mm2 images associated with low apparent diffusion coefficient (ADC) values. By age 6 months, the lesions became more prominent on b = 1000 s/mm2 images with lower ADC values. Diffusion MR imaging findings likely reflected the histopathologic changes of the disease which consisted of spongiosis of the myelinated brain tissue due to myelin vacuolation. PMID- 12886138 TI - Epidermal nevus syndrome: megalencephaly with bihemispheric and cerebellar involvement: imaging and neuropathologic correlation. AB - Epidermal nevus syndrome (ENS) is a sporadic neurocutaneous disorder characterized by the combination of epidermal nevi with significant central nervous system, eye, and skeletal abnormalities. Hemimegalencephaly and related brain abnormalities are the dominant neuropathologic abnormalities in ENS. Magnetic resonance imaging features of the case reported here revealed enlargement of both cerebral hemispheres and malformed basal ganglia, with incomplete separation of the caudate nucleus and putamen. In addition, the right cerebellar hemisphere was enlarged, with disorganized folia. None of these findings have been reported previously in this disorder. PMID- 12886137 TI - Uncommon magnetic resonance imaging observation of lumbar subdural hematoma with cranial origin. AB - A patient with a history of pain, paresthesias, and weakness in both legs is reported. Magnetic resonance imaging (MRI) of the lumbar spine demonstrated subacute subdural hematoma. Brain MRI obtained 1 day later because of progressive headache showed hemorrhagic cortical metastasis and extensive subdural hematoma. It is hypothesized that the lumbar hematoma originated from the intracranial bleeding, which was substantiated by the observation of a thin hemorrhagic collection connecting cranial and lumbar hematomas on MRI. Cranial origin should be included in the differential diagnosis of spontaneous spinal subdural hematomas. PMID- 12886140 TI - Phenylketonuria: diffusion magnetic resonance imaging and proton magnetic resonance spectroscopy. AB - Two patients with phenylketonuria are reported with white matter lesions. Diffusion magnetic resonance (MR) imaging revealed restricted diffusion patterns (high signal) on b = 1000 s/mm2 images associated with low apparent diffusion coefficient values ranging between 0.44 x 10-3 mm2/s and 0.56 x 10-3 mm2/s. On proton MR spectroscopy obtained in 1 of the patients, a prominent peak resonating at approximately 3.80 ppm was consistently present attributable to the peak rising from the alpha-proton of the phenylalanine molecule. PMID- 12886141 TI - Congenital cleft in the pontomedullary junction. AB - A 4-month-old boy is reported with complete absence of horizontal gaze. Magnetic resonance imaging revealed a congenital cleft at the pontomedullary junction directed from the right anterior paramedian region toward the posterior midline, sparing the posterior region of the pontine tegmentum and the medulla oblongata. Such clefts are extremely rare, and there is only 1 previous report in the literature. PMID- 12886142 TI - Magnetic resonance imaging and spectroscopy in a patient with treated methylmalonic acidemia. AB - Magnetic resonance imaging (MRI) and proton magnetic resonance spectroscopy (proton MRS) findings before and after carnitine therapy in a patient with methylmalonic acidemia (MMA) are evaluated. The hyperintensity on T2-weighted image and diffusion-weighted image and the decreased apparent diffusion coefficient of bilateral basal ganglia were normalized in parallel with normalization of the elevated lactate levels and the decreased N-acetyl-aspartate levels, corresponding to improvement of clinical symptoms. MRI and proton MRS may serve as a suitable, noninvasive modality for monitoring treated MMA. PMID- 12886143 TI - Reversible MR imaging and MR spectroscopy abnormalities in association with metronidazole therapy. PMID- 12886144 TI - Bladder imaging using multidetector row computed tomography, volume rendering, and magnetic resonance imaging. AB - Both multidetector row computed tomography (MDCT) and magnetic resonance imaging (MRI) are used to evaluate the bladder noninvasively. MDCT offers fast imaging with near-isotropic data sets optimized for three-dimensional imaging, including the latest software for volume rendering. MRI provides distinctive soft tissue contrast resolution and can perform dynamic imaging without radiation exposure. This article discusses the techniques and protocols of each modality with case illustrations of their application in a range of bladder pathologies to show their respective distinct advantages and limitations. PMID- 12886145 TI - Magnetic resonance imaging findings in gynecologic emergencies. AB - Gynecologic emergencies include a variety of gynecologic diseases that manifest with acute lower abdominal pain, fever, and vaginal bleeding. Recent technical advances enable the application of magnetic resonance (MR) imaging to these conditions, which may add clinically useful information in a short enough time for emergent diagnosis. In this study, the acute gynecologic conditions in which MR imaging studies were performed are reviewed, and the characteristic MR findings in gynecologic emergencies, including ovarian hemorrhage, ectopic pregnancy, tumor rupture, torsion, hemorrhage, infarction, and pelvic inflammatory diseases, are demonstrated. Familiarity of these findings aids accurate diagnosis and proper treatment choices in gynecologic emergencies. PMID- 12886146 TI - Magnetic resonance imaging of the liver: true fast imaging with steady state free precession sequence facilitates rapid and reliable distinction between hepatic hemangiomas and liver malignancies. AB - OBJECTIVE: To assess the capability of the true fast imaging with steady state free precession (true FISP) sequence in the distinction between hemangiomas and malignant liver lesions. METHODS: Sixty-eight patients with 45 hemangiomas and 51 liver malignancies were included in this study. A 1.5-T magnetic resonance system and a phased-array body coil were used. In addition to true FISP, breath-hold and fat-suppressed, T2-weighted, half-Fourier single-shot turbo spin echo (HASTE) and both unenhanced and gadolinium (Gd)-enhanced T1-weighted sequences were acquired. Two radiologists evaluated the magnetic resonance images independently in a blinded fashion. Interobserver variations with true FISP and HASTE were determined. Lesion contrast-to-noise ratios were calculated from true FISP images. RESULTS: With true FISP, readers 1 and 2 made a correct distinction between hemangiomas and liver malignancies in 43 of 45 (96%) cases and 40 of 45 (89%) cases, respectively. The kappa value was 0.65. With HASTE, the success rates were 40 of 45 cases (89%) and 36 of 45 cases (80%), respectively, and the kappa value was 0.33. With a Gd-enhanced T1-weighted sequence, the correct classifications were 35 of 45 cases (78%) and 37 of 45 cases (82%), respectively. All hemangiomas appeared as bright and well-demarcated lesions on true FISP images. Malignant liver foci were heterogeneous with unsharp margins and nearly isointense relative to liver. The specificities of true FISP in lesion differentiation were 100% and 98% for readers 1 and 2, respectively. The mean contrast to noise ratio value of hemangiomas was 21.2 (standard deviation [SD] = 9.2), and that of malignant lesions was 4.9 (SD = 3.9). This difference was highly significant (P < 0.0001). CONCLUSION: Noninvasive, rapid, and reliable differentiation between hemangiomas and malignant liver lesions is possible by using the true FISP sequence. PMID- 12886147 TI - Disconnected pancreatic duct syndrome: imaging findings and therapeutic implications in 26 surgically corrected patients. AB - PURPOSE: The lack of ductal continuity between a viable pancreatic tissue and the gastrointestinal tract results in the disconnected pancreatic duct syndrome (DPDS). The purpose of our study is to describe accurately the imaging features of CT scanning and endoscopic retrograde pancreatography (ERCP) that define the DPDS. METHODS: We conducted a retrospective analysis of the computed tomography (CT) and ERCP examinations in 26 consecutive patients with surgically proven disconnected pancreatic ducts treated over a 5-year period at our institution. Two abdominal radiologists concurrently defined the imaging features (presence and size of fluid collection along the course of the pancreatic duct, upstream enhancing pancreatic parenchyma, and ERCP abnormalities) via consensus for both exams. Patient demographics, etiology of pancreatitis, surgical treatment, initial CT interpretation, and the delay between symptom onset to correct diagnosis were recorded. RESULTS: A discrete, intrapancreatic fluid collection (average size = 27 cm2 (range, 4-74 cm2) along the course of the main pancreatic duct with upstream viable pancreatic parenchyma was identified by CT in 26 cases. ERCP showed ductal obstruction at the level of the intrapancreatic fluid collection in all patients with extravasation of contrast in 14 (54%). All patients were treated by operation: 15 (58%) by internal drainage into a Roux-en Y limb of jejunum and 11 (42%) by distal pancreatic resection. No prior CT interpretation correctly identified DPDS. The average delay between symptom onset and definitive diagnosis was 9.3 months (range, 3-36 months). CONCLUSIONS: A discrete intrapancreatic fluid collection along the expected course of the main pancreatic duct with viable upstream pancreatic parenchyma suggests the diagnosis of DPDS. ERCP findings of ductal obstruction at the level of this fluid collection with or without contrast extravasation confirm this diagnosis. Treatment is surgical and requires either internal drainage or distal pancreatic resection for complete resolution. PMID- 12886148 TI - Patient gender-related performance of nonfocused helical computed tomography in the diagnosis of acute appendicitis. AB - OBJECTIVE: To determine the impact of patient gender on the performance of helical computed tomography (CT) in the diagnosis of acute appendicitis. MATERIALS AND METHODS: From January 1, 1996 to December 31, 2000, 650 consecutive nonfocused helical abdominal CT scans were performed in adult patients presenting with acute lower abdominal pain. In general, most patients received both intravenous and oral contrast with 5-mm scan collimation through the lower abdomen and pelvis; details regarding technique and overall accuracy have been published previously. We subanalyzed results with respect to patient gender related differences, especially in false-positive and false-negative cases. A chi2 analysis was performed to determine if significant gender-related differences were present in major or minor CT diagnostic criteria for acute appendicitis, extra-appendiceal findings, sensitivity, specificity, and accuracy. RESULTS: Of the 650 patients, 552 had adequate clinical follow-up. The sensitivity for diagnosing acute appendicitis was 100% (65/65) in men and 93.6% (74/79) in women (P < 0.05); specificity was 96.2% (130/135) in men and 98.9% (272/275) in women (P > 0.05); and accuracy was 97.5% (193/198) in men and 97.6% in women (346/354) (P > 0.05). A thickened appendix and periappendiceal stranding were seen in 92.1% of men and 84.5% of women (P = 0.15) All 5 false-negative results were in thin women. In patients without acute appendicitis, CT was able to provide a relevant alternative diagnosis in 183 of 272 women (67.3%) and 81 of 130 men (62.3%) (P > 0.05). CONCLUSION: Nonfocused helical CT was highly accurate for diagnosing acute appendicitis in both men and women, although there was a slight but significant decrease in sensitivity in thin women. PMID- 12886149 TI - Computed tomography demonstration of immediate and delayed complications of computed tomography-guided transthoracic percutaneous ethanol injection of hepatocellular carcinoma at the liver dome. AB - OBJECTIVE: We conducted this study to correlate computed tomography (CT) images with the complications induced by CT-guided percutaneous ethanol injection (PEI) for the treatment of hepatocellular carcinoma (HCC). METHODS: Twenty-two patients with ultrasonographically unreachable HCC were treated with CT-guided transthoracic PEI. The procedures and the CT findings were recorded to correlate with the various complications of PEI. RESULTS: After the ethanol injection, 3 patterns of CT images were identified according to the distribution of ethanol (ie, intratumoral, extrahepatic, and intrahepatic patterns). The intrahepatic pattern was further classified as the branch, tubular, and infiltrative types. During the follow-up period, 3 patients developed biliary stricture, portal vein thrombosis, or tumor seeding. The tubular type was associated with the development of biliary stricture, whereas the branch type was related to the occurrence of portal vein thrombosis. These complications were considered to be caused by a greater amount of ethanol leakage into bile ducts or portal veins than in cases without complications (median = 17 mL vs. 0 mL, P = 0.002). CONCLUSIONS: Ethanol leakage is not uncommon when performing PEI; however, it seems to be safe when the leakage is less than 15 mL. Injection of a small amount of ethanol or monitoring by CT may be helpful in preventing the development of these complications. PMID- 12886150 TI - Accurate determination of metabolic rates from dynamic positron emission tomography data with very-low temporal resolution. AB - PURPOSE: The graphical approach is widely used for the pixelwise determination of local metabolic rate of glucose from dynamic positron emission tomography (PET) data. In its conventional implementation, measured integrals over time frames are used to approximate instantaneous tracer concentrations at midframe times ("midframe approach"). This is justified in case of high temporal resolution of the PET measurement; that is, if scan protocols with a large number of short frames are used. This requires fast data handling and large amounts of memory. Cardiac gating and three-dimensional (3D) acquisition of dynamic studies is hardly possible with this approach. Therefore, a new variant of the graphical method is proposed which can be used with a very low number of rather long frames. METHODS: An operational equation of the graphical method was derived which uses measured time integrals only and, thus, avoids the systematic errors of the midframe approximation. This "integral approach" was evaluated in computer simulations based on experimental data. RESULTS: The integral approach enables the use of protocols with 3 frames only without compromising accuracy of the derived metabolic rates whereas the midframe approach leads to bias of about 10% to 20% for these protocols. Furthermore, test-retest stability can significantly be improved when using the integral approach. CONCLUSION: The integral approach to the graphical evaluation of dynamic PET data yields accurate and precise results using scan protocols with down to only 3 frames. This can be relevant to gating and/or 3D acquisition of dynamic studies. The integral approach is applied most naturally whenever the input function is derived from the dynamic PET data. PMID- 12886151 TI - Adrenal carcinoma with a signal loss on chemical shift magnetic resonance imaging. AB - A case of primary adrenal carcinoma that showed a decreased signal in part of the tumor on chemical shift magnetic resonance imaging (CSI) is reported. The patient had clinical symptoms of Cushing syndrome. The tumor was a discrete right adrenal mass about 3.4 cm in largest diameter. It was surgically removed and diagnosed as adrenal carcinoma histopathologically. Two portions where clear cells were prominent were recognized. These areas probably contributed to the signal loss on CSI. PMID- 12886152 TI - Gastrointestinal stromal tumor of rectum: a report of 2 cases. AB - The specific c-kit protein (CD117)-positive gastrointestinal stromal tumor (GIST) of the rectum has been sporadically reported in pathology literature, but its radiologic features have never been thoroughly described. We present 2 cases of histologically and immunohistochemically confirmed rectal GISTs. Their clinical and multimodality imaging features are illustrated, and the relevant literature is reviewed and discussed. PMID- 12886153 TI - Pancreatic herniation through the gastroesophageal hiatus: magnetic resonance imaging, magnetic resonance cholangiopancreatography, and computed tomography evaluation. AB - A rare case of pancreatic herniation through the gastroesophageal hiatus is presented, including magnetic resonance evaluation with magnetic resonance cholangiopancreatography and magnetic resonance angiography images unique to the published literature. PMID- 12886154 TI - Multidetector row computed tomography and volume rendering of an adult duodenal intraluminal "wind sock" diverticulum. AB - Adult duodenal intraluminal "wind sock" diverticulum (IWD) is a rare entity commonly complicated by pancreatitis and usually diagnosed by barium study or at surgery. We present surgical correlation of a case studied with multidetector row computed tomography (MDCT) and three-dimensional volume rendering (3DVR), which offered some unique insight into this condition and its pathophysiology. PMID- 12886155 TI - Localized pseudomyxoma peritonei in the female pelvis simulating ovarian carcinomatous peritonitis. AB - Two cases of localized pseudomyxoma peritonei in the female pelvic cavity associated with a ruptured appendiceal mucocele and ovarian involvement that mimicked ovarian carcinomatous peritonitis were evaluated. Subtle omental irregularity adjacent to the cecum may suggest the hidden appendiceal origin reflecting localized carcinomatous peritonitis caused by the occult rupture of the mucocele. Mucinous fluid-like materials were localized in the pelvic cavity with scalloping of the uterus, which may be the diagnostic finding of pseudomyxoma peritonei. PMID- 12886156 TI - Diffuse hepatic angiosarcoma with a portal venous supply mimicking hemangiomatosis. AB - We report a case of diffuse hepatic angiosarcoma. Dynamic computed tomography (CT) showed gradual contrast enhancement and homogeneity in the late phase. The tumor was homogeneously hypointense on T1-weighted imaging and hyperintense on T2 weighted imaging, mimicking hemangiomatosis. An angiogram demonstrated intrahepatic arterial encasement, which suggested malignancy. The portal venous supply to the tumor was demonstrated by CT during arterioportography. This probably reflected the continuous infiltration of atypical cells into the dilating sinusoids and may also suggest malignancy. PMID- 12886157 TI - Value of magnetic resonance imaging in the depiction of intravenous leiomyomatosis extending to the heart. AB - Intravenous leiomyomatosis is a seldom neoplasia characterized by invasion of venous channels by a benign smooth muscle tumor originating either from a uterine myoma or from vessel wall. Extension to the heart may cause mechanical obstruction and is frequently misdiagnosed as a right-atrial myxoma. We present a case of recurrent intravenous leiomyomatosis with previous hysterectomy because of uterine leiomyoma which have different magnetic resonance characteristics than that of the former reports. PMID- 12886158 TI - Parachordoma with multiple metastases. AB - We present a very rare case of parachordoma with local aggressiveness and widespread metastases. A 68-year-old male presented with a mass in his left calf. The lesion was depicted as a poorly marginated mass with inhomogeneous signal intensity on magnetic resonance imaging. The tumor invaded surrounding muscles, neurovascular bundles, and bones. Widespread metastasis to lung, bone, and skin developed after amputation surgery. The histologic features of primary and metastatic lesions were the same and consistent with parachordoma. PMID- 12886159 TI - Neural fibrolipoma of the foot. AB - Neural fibrolipoma is a benign tumor comprised of hypertrophied fibrofatty tissue with intermixed nerve tissue. We present the case of a neural fibrolipoma of the foot that underwent above-ankle amputation and review the characteristic features of this unique form of localized gigantism. PMID- 12886160 TI - Interstitial magnetic resonance lymphography: comparative animal study of gadofluorine 8 and gadolinium diethylenetriamine-pentaacetic acid. AB - PURPOSE: We evaluated the usefulness of gadofluorine 8 (G8) and gadolinium diethylenetriamine-pentaacetic acid (Gd-DTPA) for interstitial magnetic resonance lymphography (MRL). METHOD: Phantom imaging studies were conducted with G8 and Gd DTPA corresponding to 0.1-0.005 micromol Gd/mL, and signal intensity was measured. Popliteal lymph node (PLN) accumulation was investigated in rabbits. Imaging was performed before and after subcutaneous administration of G8 (50 micromol Gd/mL) and Gd-DTPA (50 micromol Gd/mL). Contrast enhancement ratio measurements of PLN were determined both prior to and following the administration. RESULTS: Signal intensity of phantom for G8 are higher than those for Gd-DTPA from 0.1 to 0.002 micromol Gd/mL (P < 0.0001). Imaging after 5 minutes can clearly detect PLN accumulation on two contrast agents. Nevertheless, PLN accumulation of Gd-DTPA disappeared after about 30 minutes. A statistically significant difference between G8 and Gd-DTPA can be seen from 5 to 90 minutes in CER (P < 0.0001). PLN became clearly visible at 5 minutes after the injection of G8 and Gd-DTPA. Nevertheless, PLN accumulation of Gd-DTPA disappeared at 30 minutes. Otherwise, PLN accumulation of G8 continued up to 90 minutes. CONCLUSION: These studies indicated that G8 was more suitable than Gd-DTPA as a contrast agent for interstitial MRL up to 90 minutes. PMID- 12886161 TI - Quantitative multilevel mapping of hepatic blood flow by xenon computed tomography using aorta. AB - A noninvasive and quantitative technique has been developed to measure human hepatic blood flow by xenon computed tomography (Xe-CT). Accurate data on time dependent xenon concentrations in the arterial blood are indispensable for Xe-CT to ensure quantitativeness of measured blood flow. A method has been established by our group to use both aorta and end-tidal data to obtain arterial xenon information. Multilevel (3 levels) maps of arterial blood flow (Fa), portal blood flow (Fp), and partition coefficient (lambda) were created for patients with chronic hepatitis. A method to objectively evaluate Fa, Fp, and lambda values for the whole liver has also been developed by our group. PMID- 12886162 TI - Incremental value of advanced image processing of multislice computed tomography data in the evaluation of hypervascular liver lesions. AB - Multiplanar volume rendering and maximum intensity projection of multislice computed tomography data allow for robust evaluation of the hepatic parenchyma, with early detection of neovascularity and tumor stain, and enable better assessment of the tumor burden compared with routine axial images. Lesion characterization can also potentially improve. This is in addition to accurate delineation of the hepatic vascular anatomy, which allows for segmental localization and assists in surgical and nonsurgical planning. PMID- 12886163 TI - The source and direction of thoracic lymphatics, part II: The lower thorax. AB - The purpose of this set of 2 articles is to illustrate the source and direction of lymphatic drainage in the thorax on axial computed tomographic (CT) schematics. The first article describes the upper thorax. This article describes the lower thorax. The nodal groups are given conventional anatomic names and the corresponding terminology of the American Thoracic Society. Arrows indicate the direction of the flow. The region or organ drained is color-coded, and nodes that receive lymph from each area are assigned appropriately colored and numbered boxes. Major drainage patterns are also described. This information can be used to assess a suspicious lymph node by tracing it to the region of drainage and looking for pathology. Alternately, one may extrapolate the potential drainage routes of a tumor and scrutinize specific sites for nodal metastases. PMID- 12886164 TI - A guest editorial: the mentor-mentee relationship. PMID- 12886165 TI - Performance of visual inspection with acetic acid for cervical cancer screening: a qualitative summary of evidence to date. AB - Developing countries often lack the necessary resources to use the Papanicolaou (Pap) smear as a screening tool for cervical abnormalities. Because the burden of cervical cancer is highest in such low-resource settings, alternative techniques have been sought. Recently, interest in visual inspection with acetic acid (VIA) has increased. Numerous studies have been conducted on its accuracy and its ability to detect cervical lesions when compared with other techniques, both conventional and nonconventional. This review summarizes key findings from the literature to provide researchers and policymakers with an up-to-date summary on VIA. PubMed was used to identify relevant journal articles published between 1982 and 2002. Key words were cervical cancer screening, visual inspection, VIA (visual inspection with acetic acid), DVI (direct visual inspection), AAT (acetic acid test), and cervicoscopy. Studies were eligible for review only if they involved analysis of primary VIA data (ie, not review articles); studies involving magnification devices were excluded. Fifteen studies were reviewed in total; key results were extracted and a summary analysis was performed for sensitivity and specificity parameters. When reported, sensitivity ranged between 66% and 96% and specificity between 64% and 98%. Authors comparing VIA with cytology noted that the overall usefulness of VIA compares favorably with that of the Pap test. The reported findings reviewed here suggest that VIA has the potential to be a cervical cancer screening tool, especially in low resource settings. TARGET AUDIENCE: Obstetricians & Gynecologists, Family Physicians. LEARNING OBJECTIVES: After completion of this article, the reader will be able to describe how visual inspection of the cervix for cervical cancer screening (VIA) is performed, to summarize the current literature on VIA, and to list potential advantages of VIA. PMID- 12886166 TI - Pregnancy following endometrial ablation: case history and literature review. AB - A total of 43 pregnancies that occurred subsequent to endometrial ablation were reported to July 2002. Only 17 of these pregnancies had progressed beyond 20 weeks. We report a successful planned pregnancy following endometrial ablation and sterilization reversal, culminating in vaginal birth after a previous Caesarean section. The English literature has been reviewed to ascertain family planning practices, uterine cavity assessment and pregnancy outcomes after endometrial ablation/resection. Pregnancy management recommendations are provided. TARGET AUDIENCE: Obstetricians & Gynecologists, Family Physicians. LEARNING OBJECTIVES: After completion of this article, the reader will be able to list the various pregnancy complications associated with a history of endometrial ablation, and to describe the changes in the endometrial cavity following endometrial ablation. PMID- 12886167 TI - Steps across the gap: tools, trials and data. PMID- 12886168 TI - A new framework for describing and quantifying the gap between proof and practice. AB - BACKGROUND: Substantial gaps often exist between every day practice and best practice as defined by research evidence. We present a framework for defining, analyzing, and quantifying such proof-to-practice gaps. METHOD: An intervention's use can be plotted over time as ideal and actual uptake curves among candidates and noncandidates. Gaps of underuse are deviations from ideal uptake among candidates and can be quantified as underuse NNPs (Number Not Prevented): the number of disease events each year that would have been prevented, but were not, because of underuse among candidates of the intervention. Gaps of overuse are deviations from ideal uptake among non candidates and can be similarly quantified as overuse NNPs. RESULTS: Applying our method to the underuse of beta-blockers at hospital discharge postmyocardial infarction (MI) in the United States demonstrates an annual NNP of 2995 first-year post-MI deaths not prevented (sensitivity analysis range 455-20,409). Our NNP analysis framework highlights challenges to the determination of efficacy and efficiency, the definition of what constitutes proof, rapid recognition of proof when it does occur, the definition of eligible candidates, and the definition of the proportion of candidates treated. CONCLUSION: League tables of NNPs can help policy makers compare the clinical consequences of underuse and overuse of diverse interventions, while the NNP framework provides a systematic approach for describing and analyzing the components of proof-to-practice gaps. Such gap analyses can help organizations direct their resources to reducing gaps of greatest clinical consequence. PMID- 12886169 TI - Differences in length of stay in Veterans Health Administration and other United States hospitals: is the gap closing? AB - OBJECTIVES: Compare risk-adjusted length of stay (LOS) in VA and other United States (non-VA) hospitals and determine if relative differences in LOS have changed in recent years. RESEARCH DESIGN: Retrospective cohort study. PATIENTS: Patients with ten common medical diagnoses admitted to all VA hospitals and to non-VA hospitals included in the National Hospital Discharge Survey (NHDS) during 1996 through 1999. DATA: Comparable data elements were obtained from VA administrative databases and the NHDS. LOS was adjusted for age, gender, marital status, and comorbidity. Comorbidity was assessed using a validated methodology that considers 30 conditions. RESULTS: Unadjusted mean LOS was longer in VA than non-VA patient for all 4 years, in aggregate (7.1 vs. 4.9 days, respectively; P < 0.001), and for each year individually. However, the difference in mean LOS in VA and non-VA patients declined from 2.9 days in 1996 to 1.6 days in 1999. LOS in VA patients remained longer (P < 0.001) in linear regression analyses, adjusting for demographics and comorbidity. However, the difference in LOS declined from 28.5% (95% CI, 28.1%-29.0%) in 1996 to 17.0% (95% CI, 16.6%-17.4%) in 1999. These results were similar in analyses of individual geographic regions. CONCLUSIONS: Risk-adjusted LOS was longer in VA hospitals than in other United States hospitals. However, differences in LOS narrowed between 1996 and 1999. These findings suggest that changes in the organization and delivery of VA health care in the mid-1990s may be closing the gap between the VA and other healthcare systems in hospital utilization. PMID- 12886170 TI - Physician and practice characteristics associated with the early utilization of new prescription drugs. AB - BACKGROUND: Prescription of new drugs contributes to substantial increases in annual drug expenditures. A small proportion of physicians appear to be early users of new prescription drugs and little is known about their characteristics. OBJECTIVE: To estimate the initial utilization rate of new prescription drugs among physicians, and the physician and practice characteristics associated with early use. DESIGN: Cumulative prospective assessment over a 5 year period (1989 1994) of new drug utilization rates in a randomly selected cohort of Quebec physicians. PARTICIPANTS: 1661 physicians and 669,867 elderly patients. OUTCOME: Prescribing rate of 20 new drugs, in 6 therapeutic categories, to elderly patients in the first 6 months after inclusion in the Quebec formulary. RESULTS: The 20 new drugs were prescribed by 1.3-22.3% of physicians, and there was an 8 to 17-fold difference in new drug utilization rates among prescribers. Characteristics associated with higher rates of utilization differed for general practitioners and specialists. Male general practitioners, and physicians graduating from the most recently established medical school in the province, had higher rates of new drug utilization, whereas recent graduation was only associated with higher utilization rates among specialists. Practice volume was associated with higher rates of utilization among GPs. For both GPs and specialists, having a high proportion of elderly in one's practice and a rural or remote practice location was associated with lower utilization rates. CONCLUSIONS: Physician sex, specialty, medical school, years since graduation, practice location, volume, and relative proportion of elderly in the physician's practice influence the utilization of new drugs. PMID- 12886171 TI - The minimum data set urinary incontinence quality indicators: do they reflect differences in care processes related to incontinence? AB - PURPOSE: To determine if nursing homes that score in the lower 25th percentile (low prevalence) versus the upper 75th percentile (high prevalence) on each of two Minimum Data Set (MDS) incontinence quality indicators provide different incontinence care processes DESIGN: Cross-sectional. SUBJECTS: 347 long-term residents in 14 skilled nursing facilities for the MDS "prevalence of incontinence" indicator and 432 residents in 16 skilled nursing facilities for the MDS "prevalence of incontinence without a toileting plan" indicator. MEASURES: Nine care processes related to incontinence were defined and operationalized into clinical indicators. Research staff assessed implementation of each care process on 3 consecutive 12-hour days (7 am to 7 pm ). The assessment included resident interviews, physical performance evaluations, and chart abstraction using standardized protocols. RESULTS: Homes with lower prevalence rates on both MDS incontinence quality indicators (good outcomes) had a significantly higher proportion of participants with chart documentation of two relevant care processes: 1 an evaluation of the resident's incontinence history and 2 toileting assistance rendered by staff. However, interviews with incontinent residents capable of accurately reporting care activity occurrence showed no difference in toileting assistance frequency between homes in the upper and lower quartiles for either MDS incontinence indicator. Participants reported an average of 1.8 toileting assists per day across all homes with a narrow average frequency range between homes (1.6-2.0). These frequencies of toileting assistance are not sufficient to improve urinary incontinence. There was also no difference in the frequency of toileting assistance received by incontinent participants rated on the MDS as receiving scheduled toileting (n = 75, mean = 1.9 +/- 1.24) compared to incontinent residents rated on the MDS as not receiving scheduled toileting (n = 131, mean = 1.8 +/- 1.22). None of the homes provided chart documentation that supported staff decisions to place or not place a resident on a scheduled toileting program. CONCLUSIONS: The quality of incontinence assessment and treatment as documented by scheduled toileting interventions was poor across all homes, and the MDS incontinence quality indicators were not associated with clinically important differences in related care processes. Chart documentation that a resident was on a scheduled toileting program or received toileting assistance was not related to resident reports of the frequency of received assistance. PMID- 12886172 TI - Measuring patients' perceptions of the outcomes of treatment for early prostate cancer. AB - BACKGROUND: Compared with careful attention to the physical (eg, urinary, bowel, sexual) dysfunction that may follow treatment, little attention has been given to the behavioral, emotional, and interpersonal changes that the diagnosis of early prostate cancer and subsequent physical dysfunction may bring. OBJECTIVE: To construct patient-centered measures of the outcomes of treatment for early prostate cancer. RESEARCH DESIGN: Qualitative study followed by survey of early prostate cancer patients and group of comparable patients with no history of prostate cancer. Analysis of focus groups identified relevant domains of quality of life, which were represented by Likert scale items included in survey questionnaires. Psychometric analyses of survey data defined scales evaluated with respect to internal consistency and validity. RESULTS: Qualitative analysis identified three domains: urinary control, sexuality, and uncertainty about the cancer and its treatment. Psychometric analysis defined 11 scales. Seven were generically relevant to most older men: urinary control (eg, embarrassment with leakage), sexual intimacy (eg, anxiety about completing intercourse), sexual confidence (eg, comfort with sexuality), marital affection (eg, emotional distance from spouse/partner), masculine self esteem (eg, feeling oneself a whole man), health worry (eg, apprehensiveness about health changes), and PSA concern (eg, closely attending to one's PSA). Four scales were specific to the treatment experience: perceived cancer control, quality of treatment decision making, regret of treatment choice, and cancer-related outlook. CONCLUSION: The scales provide definition and metrics for patient-centered research in this area. They complement measures of physical dysfunction and bring into resolution outcomes of treatment that have gone unnoticed in previous studies. PMID- 12886173 TI - Tooth retention as an indicator of quality dental care: development of a risk adjustment model. AB - BACKGROUND: Retaining teeth improves oral health and quality of life. Thus, receipt of a root canal (vs. a tooth extraction) is a useful indicator of the quality of dental care. However, use of this quality measure without adjusting for the severity of oral disease could lead to spurious conclusions. OBJECTIVES: This paper describes the development of a dental severity adjustment methodology. RESEARCH DESIGN: Retrospective study. SUBJECTS: 54,423 users of Department of Veterans Affairs (VA) dental care who had either root canal therapy or a tooth extraction at a VA facility in Fiscal year 1998. MEASURES: International Classification of Disease Clinical Modification codes for dental diagnoses and comorbid medical conditions. We modeled the effects of dental disease severity in logistic regression models of the probability of receiving a root canal, using both conceptual and Modified Delphi-Panel derived models, adjusting for age, and medical comorbidities. RESULTS: Conceptual and Modified Delphi models performed similarly. The dental disease severity adjustments increased the fit in models of the probability of receiving a root canal (C-statistic = 0.822 for the conceptual model and 0.804 for the Modified Delphi Panel model) compared with the model including comorbid medical conditions alone (C-statistic = 0.561). CONCLUSIONS: Risk adjustment for dental disease severity improves the fit of models of the probability of receiving a root canal. Studies of the quality of dental care should consider employing risk-adjusted models. PMID- 12886174 TI - Gender Differences in service connection for PTSD. AB - BACKGROUND: Mentally ill female veterans obtain a smaller proportion of their care from Department of Veterans Affairs (VA) facilities than mentally ill male veterans do, possibly because women are less likely than men to be service connected for psychiatric disabilities. "Service connected" veterans have documented, compensative conditions related to or aggravated by military service, and they receive priority for enrollment into the VA healthcare system. OBJECTIVES: To see if there are gender discrepancies in rates of service connection for posttraumatic stress disorder (PTSD) and, if so, to see if these discrepancies could be attributed to appropriate subject characteristics (eg, differences in symptom severity or impairment). RESEARCH DESIGN: Mailed survey linked to administrative data. Claims audits were conducted on 11% of the sample. SUBJECTS: Randomly selected veterans seeking VA disability benefits for PTSD. Women were oversampled to achieve a gender ratio of 1:1. RESULTS: A total of 3337 veterans returned usable surveys (effective response rate, 68%). Men's unadjusted rate of service connection for PTSD was 71%; women's, 52% (P < 0.0001). Adjustment for veterans' PTSD symptom severity or functional impairment did not appreciably reduce this discrepancy, but adjustment for dissimilar rates of combat exposure did. Estimated rates of service connection were 53% for men and 56% for women after adjusting for combat exposure. This combat preference could not be explained by more severe PTSD symptoms or greater functional impairment. CONCLUSIONS: Instead of a gender bias in awards for PTSD service connection, we found evidence of a combat advantage that disproportionately favored men. The appropriateness of this apparent advantage is unclear and needs further investigation. PMID- 12886175 TI - Comparing the cost-effectiveness of the triple test score to traditional methods for evaluating palpable breast masses. AB - BACKGROUND: Physical examination, mammography, ultrasonography, and fine needle aspiration are traditionally used to guide further management of palpable breast masses, often leading to open biopsy of benign masses. The triple test score (TTS) integrates physical examination, mammography, and fine needle aspiration in the initial evaluation, limiting open biopsy. OBJECTIVE: To compare cost effectiveness of TTS and traditional methods. METHODS: The primary measure of clinical effectiveness, frequency of missed malignancy, was determined for each strategy using probabilities and outcomes from a systematic literature review. Costs were calculated using the Medicare resource-based relative-value scale. A decision-analytic model compared costs of initial work-up, costs per mass evaluated, and costs per malignancy diagnosed. Sensitivity analyses assessed the influence of variations in model assumptions. RESULTS: In the base case, neither strategy led to undiagnosed breast cancer. However, open biopsy was required in 13% of benign masses using TTS versus 88% using the traditional strategy. The cost of the initial work-up using traditional management was less than TTS (377 US dollars vs. 627 US dollars), but cost per mass evaluated and cost per malignancy diagnosed (1793 US dollars vs. 925 US dollars and 5670 US dollars vs. 2925 US dollars) favored TTS, due to substantially reduced open biopsy.In sensitivity analyses, TTS cost varied most with changes in cost of initial evaluation, whereas the traditional strategy cost varied most with changes in open biopsy cost. CONCLUSIONS: The TTS provides equivalent diagnostic effectiveness but substantially lower cost than traditional management. Cost savings are based on decreased open biopsy, a major contributor to the cost of traditional evaluation in this model. PMID- 12886176 TI - How does beneficiary knowledge of the Medicare program vary by type of insurance? AB - BACKGROUND: Prior research found that Medicare beneficiaries' knowledge of the Medicare program varied by the type of supplemental insurance they had. However, none of these studies used both multivariate methods and nationally representative data to examine the issue. OBJECTIVES To measure beneficiary knowledge of the Medicare program and to evaluate how knowledge varies by type of supplemental insurance. RESEARCH DESIGN: A mail survey with telephone follow-up to a nationally representative random sample of Medicare beneficiaries, which had a 76% response rate. The purpose of the study was to evaluate the effects of providing the Medicare & You handbook on beneficiary knowledge, information needs, and health plan decision making. SUBJECTS: A total of 3738 Medicare beneficiaries who completed the survey. MEASURES: A psychometrically validated 22 item index that reflects Medicare-related knowledge in seven different content areas. RESULTS Overall, beneficiaries with a Medicare HMO or non-employer sponsored supplemental insurance were more knowledgeable about Medicare than those who had Medicare only. In general, beneficiaries tended to be more knowledgeable about issues related to the type of insurance they had (fee-for service or managed care) than other types of insurance. CONCLUSIONS: Higher levels of knowledge about one's own type of insurance may suggest that beneficiaries learn by experience or they learn more about that type of insurance before enrollment. Further research is needed to better understand how and when beneficiaries learn about insurance and what educational strategies are more effective at increasing knowledge. PMID- 12886177 TI - Postoperative mortality and pulmonary complication rankings: how well do they correlate at the hospital level? AB - BACKGROUND: Postoperative mortality rankings are used alone for quality assessment. OBJECTIVES: To determine the correlation between hospital rankings of postoperative respiratory failure, pneumonia, and mortality rates and to assess the influence of hospital volume, type of surgery, and time on these correlations. To compare hospital outlier detection with and without pulmonary complication rates. RESEARCH DESIGN: Prospective observational study. SUBJECTS: 103,176 noncardiac surgery patients from 123 VA hospitals enrolled between 1/1/94 and 8/31/95. Preoperative pneumonia, ventilator dependent, comatose, or do-not resuscitate patients were excluded. MEASURES: Respiratory failure was defined as greater than 48 hours of ventilator assistance or postoperative reintubation. Pneumonia was defined as positive sputum cultures with antibiotic treatment or chest x-ray infiltrate diagnosed as pneumonia or pneumonitis. Mortality was defined as death within 30 days of surgery. Hospital rankings were assigned using risk-adjusted observed-to-expected ratios. RESULTS: There was significant, but weak correlation between mortality and pulmonary complication rankings (r = 0.21, P = 0.02 for pneumonia; r = 0.22, P = 0.01 for respiratory failure). Correlations with mortality rankings were highest for thoracic (r = 0.42, P < 0.001 for pneumonia; r = 0.38, P < 0.001 for respiratory failure) and vascular surgery (r = 0.26, P = 0.02 for pneumonia; r = 0.35, P < 0.001 for respiratory failure). Supplementing mortality with pulmonary complication outlier designations enhanced outlier detection for 47% of hospitals overall, and for 29% in the lowest caseload quartile. CONCLUSIONS: Pulmonary complication rankings correlate weakly with mortality overall, but have higher correlations in thoracic, vascular, and upper abdominal surgery. Examining pneumonia and respiratory failure outlier status with mortality outlier status enhances hospital outlier detection even in low-volume hospitals. PMID- 12886178 TI - Family physician continuity of care and emergency department use in end-of-life cancer care. AB - BACKGROUND: Despite cancer patients preferring to spend their last days out-of hospital, many make difficult visits to the emergency department (ED). Family physician continuity of care has been shown in some clinical situations to reduce ED utilization. OBJECTIVE: To determine if greater family physician continuity of care for cancer patients during the end-of-life is associated with less ED utilization. METHOD: This retrospective, population-based study involved secondary analysis of linked administrative data files for 1992 to 1997. Sources included the Nova Scotia Cancer Registry, Vital Statistics, the Queen Elizabeth II Health Sciences Center Oncology Patient Information System and Palliative Care Program (PCP), Hospital Admissions/Separation data, and Physician Services information. Subjects included adults with a recorded date of cancer diagnosis who died of cancer and who had made at least three visits to a family physician during their last 6 months of life. The relationship between total ED visits and family physician continuity of care, developed using the Modified Modified Continuity Index (MMCI), was examined using negative binomial regression with adjustments for survival, year of death, sex, age, cancer type, region, PCP admission, specialty visits, hospital days, death location, income quintile, and total ambulatory visits. RESULTS: In total, 8702 subjects made 11,551 ED visits (median = 1.0); median MMCI was 0.83. Adjusted results indicate those experiencing low continuity (MMCI < 0.5) made 3.9 times more ED visits (rate ratio [RR] = 3.93; 95% CI [CI] = 3.57-4.34) than those experiencing high continuity (MMCI > or = 0.8) and patients experiencing moderate continuity (MMCI = 0.5-0.8) made twice as many ED visits (RR = 2.28; CI = 2.15-2.42). CONCLUSION: Given this significant association between family physician continuity of care and ED visits during the end-of-life, and given international trends to reform primary care, active planning of strategies to facilitate such continuity should be encouraged. PMID- 12886203 TI - Augmentation of an anterior edentulous ridge for fixed prosthodontics with combined use of orthodontics and surgery: a clinical report. AB - Untreated periodontitis may lead to tooth and tissue loss that can result in diminished masticatory function and esthetic deformities. Combined periodontal/prosthodontic treatment for patients with advanced periodontal disease has been well documented, and treatment results can often be improved with the use of adjunctive orthodontic treatment such as forced eruption. This clinical report describes a method of preprosthetic preparation of an edentulous ridge for a fixed partial denture that incorporates the combined use of orthodontics and surgical ridge augmentation. PMID- 12886204 TI - Longitudinal treatment of a severely atrophic mandible: a clinical report. AB - This clinical report presents the treatment of a patient with a severely atrophic mandible. The report details the clinical successes and failures over nearly a 20 year period. Clinical and surgical procedures are presented in a longitudinal fashion, as well as the rationale at the time to support each procedure. PMID- 12886205 TI - Clinical complications with implants and implant prostheses. AB - The purpose of this article is to identify the types of complications that have been reported in conjunction with endosseous root form implants and associated implant prostheses. A Medline and an extensive hand search were performed on English-language publications beginning in 1981. The searches focused on publications that contained clinical data regarding success/failure/complications. The complications were divided into the following 6 categories: surgical, implant loss, bone loss, peri-implant soft tissue, mechanical, and esthetic/phonetic. The raw data were combined from multiple studies and means calculated to identify trends noted in the incidences of complications. The most common implant complications (those with a greater than a 15% incidence) were loosening of the overdenture retentive mechanism (33%), implant loss in irradiated maxillae (25%), hemorrhage-related complications (24%), resin veneer fracture with fixed partial dentures (22%), implant loss with maxillary overdentures (21%), overdentures needing to be relined (19%), implant loss in type IV bone (16%), and overdenture clip/attachment fracture (16%). It was not possible to calculate an overall complications incidence for implant prostheses because there were not multiple clinical studies that simultaneously evaluated all or most of the categories of complications. Although the implant data had to be obtained from different studies, they do indicate a trend toward a greater incidence of complications with implant prostheses than single crowns, fixed partial dentures, all-ceramic crowns, resin-bonded prostheses, and posts and cores. PMID- 12886206 TI - Effects of acid hydrolysis and mechanical polishing on surface residual stresses of low-fusing dental ceramics. AB - STATEMENT OF PROBLEM: Cracks may arise in a ceramic restorative material over time, resulting in sudden fractures at stresses well below the yield stress. PURPOSE: This study evaluated by means of indentation technique the effects of acid hydrolysis and mechanical polishing on the surface residual stresses of low fusing ceramic materials. MATERIAL AND METHODS: A total of 64 ceramic bars were formed to produce 4 groups of 16 bars each for 4 ceramic materials (Duceram-LFC Dentin, Duceram-LFC Enamel, Finesse Dentin, and Finesse Enamel). Four surface treatment groups (n=4) were then formed for each of the 4 materials. The 4 surface treatments were control (autoglaze), hydrolysis, glaze/polish, and polish/glaze. A Vickers indenter contacted the Duceram-LFC specimens with a 5-N load and the Finesse specimens with a 3-N load for 10 seconds. Scanning electron microscope (SEM) was used to study surface texture before and after hydrolysis and polishing. Differences in mean crack lengths were analyzed with 1-way analysis of variance and least significant difference test (alpha=.05.) RESULTS: SEM showed obvious surface flaws as a result of hydrolysis on Duceram-LFC Enamel and Dentin specimens. However, statistical analysis of the resulting crack lengths revealed no significant differences between values for the control groups (58.16 +/- 3.88) (53.53 +/- 2.67) and hydrolysis groups (57.11 +/- 4.09) (54.54 +/- 3.15) for Enamel (P=.081) and Dentin (P=.093) respectively. When comparing polished groups and nonpolished groups, the mean crack lengths were significantly shorter for polished specimens of Duceram-LFC Enamel (53.76 +/- 3.17), Finesse Enamel (40.56 +/- 3.31), and Finesse Dentin (39.76 +/- 3.81) porcelains compared with their control groups (58.16 +/- 3.88) (43.54 +/- 4.12) (41.19 +/- 3.47), respectively (P<.0001). The mean crack lengths were significantly longer for polished specimens of Duceram-LFC Dentin (59.16 +/- 3.52) porcelain compared with the control group (53.53 +/- 2.67) (P<.0001). CONCLUSION: Within the limitations of this study, hydrolysis did not improve surface residual stresses of Duceram LFC and Finesse ceramic materials. Mechanical polishing improved surface residual stresses of all materials tested, except Duceram-LFC Dentin porcelain. PMID- 12886207 TI - The effect of tray selection, viscosity of impression material, and sequence of pour on the accuracy of dies made from dual-arch impressions. AB - STATEMENT OF PROBLEM: Dual-arch trays are often used to generate impressions of prepared teeth and of the opposing arch simultaneously. There is concern that accuracy of the casts generated with this technique can be affected by the type of tray, viscosity of the impression material, and sequence of pouring the cast. PURPOSE: This study compared the accuracy of working dies made from impressions with metal and plastic dual-arch trays, for 2 different viscosities of impression tray material and by altering which side of the impression was poured first. MATERIALS AND METHODS: Impressions were made of a typodont mandibular arch containing a circular stainless steel crown preparation (standard). There were 3 variables: type of dual-arch tray, impression material viscosity, and order of pour of the impressioned arches. A balanced design with independent samples was used (n=10). Two types of dual-arch trays, plastic (Triple Tray) and metal (COE Impression Tray), and 2 viscosities of addition silicone for the tray were used (Aquasil Rigid and Aquasil Monophase). Type IV gypsum (Fuji-Rock) with a ratio of 20 mL of distilled water to 100 g of powder was hand-mixed for 10 seconds then mixed under vacuum for 40 seconds and poured into the trays while being vibrated. One side of the dual-arch impression was poured with 35 g of stone and allowed to set for 1 hour before the other side was poured with 35 g of stone. The order of pour was randomized, and all casts were allowed to set for 24 hours at room temperature before removal. The dies were measured in 3 dimensions (buccolingual, mesiodistal, and occlusogingival) with a measuring microscope. The gypsum working dies were placed into a custom jig fabricated to permit measurement at a fixed, reproducible position under the microscope. Each dimension of the working dies was measured 3 times, and the mean was used for the sample value. The same 3 aspects of the stainless steel standard were measured multiple times, before and then at the conclusion of measuring all working dies, to arrive at the 3 standard values to which all working die means were compared. The means for the standard used in the statistical analysis were those taken at the conclusion of the study. The intraexaminer variation for measuring the standard was 0.001 mm. A 3-factor analysis of variance was used for the statistical analysis with hypothesis testing at alpha=.05. RESULTS: Statistically significant differences were found with viscosity selection for the buccolingual and occlusogingival dimensions of the working die. The rigid material produced working dies slightly taller (1 microm) than the standard, and those from the monophase material were 4 microm shorter. Regarding tray selection, metal trays were slightly more accurate in the mesiodistal dimension, and when monophase was used in a plastic tray, gypsum dies were nearly 30 microm smaller in the mesiodistal dimension (P<.05). Differences were not detected for sequence of pouring impressions. CONCLUSION: Within the limitations of this study, the monophase material, when compared with the rigid impression material, was most accurate for the occlusogingival and mesiodistal dimensions, although not as accurate in the buccolingual. This buccolingual difference (0.002 mm-0.006 mm) would be clinically inconsequential with the application of die spacer. The rigid impression material was also unaffected by tray selection for the mesiodistal, whereas monophase was affected. When a monophase impression material was used, plastic dual-arch trays yielded gypsum dies which were significantly smaller (0.029 mm) than the ones generated from the metal trays (0.006 mm). Thus rigid impression materials can be recommended for use in dual-arch trays; however, the magnitude of the differences would generally not be clinically significant because they could be compensated for with several coats of die spacer. PMID- 12886208 TI - Temperature effects on the rheological properties of current polyether and polysiloxane impression materials during setting. AB - STATEMENT OF PROBLEM: Rheological tests of elastomeric impression materials during setting have been most often conducted at room temperature rather than at intraoral temperature. Because temperature may affect properties and the setting kinetics, clinically relevant inferences may not be accurate with studies conducted at room temperature. PURPOSE: The purpose of this study was to determine the viscoelastic properties of new low- and medium-viscosity elastomeric impression materials during setting at 33 degrees C and to evaluate the medium-viscosity materials at 3 additional temperatures. MATERIAL AND METHODS: The impression materials investigated at 33 degrees C were 2 polyvinylsiloxanes (PVS) (Aquasil Deca and Aquasil LV) and 5 polyethers (PE) (Impregum Penta, Impregum Penta Soft H, Impregum Penta Soft L, Impregum Garant Soft L, and Permadyne Garant L). Three impression materials (Aquasil Deca, Impregum Penta, and Impregum Penta Soft H) were also investigated at 25 degrees, 29 degrees, and 37 degrees C. Time-dependent oscillatory rheometry was carried out on these materials (n=3) with a rheometer with a 25-mm diameter parallel plate cell. The storage modulus (G') and the loss tangent (tandelta) were determined as functions of time over a period from 0 seconds to 900 seconds, commencing 40 seconds after mixing. Induction time (t(ind)) or initial setting time and tandelta, the relative liquidlike behavior, were also computed. A single factor analysis of variance (ANOVA) was used for the properties determined at 33 degrees C and a 2-factor ANOVA was used for the temperature studies, with hypothesis testing at alpha=.05. RESULTS: The G'(t) curves for all materials displayed the expected sigmoidal shape with time, with the solid-like behavior rising slowly, then more rapidly, and again slowly to final set. The initial setting time (t(ind)) was found to be approximately 2.8 minutes for the PVS materials and for Impregum Penta and Impregum Penta Soft H, but was significantly longer for the remaining 3 PE low viscosity materials. The solid-like behavior (G') at final set or shear modulus differed among all materials, ranging from 1.0 MPa for Aquasil Decca, 1.69 for Impregum Penta Soft H, and 1.8 MPa for Impregum Penta. G' for low-viscosity materials ranged from 0.66 MPa for Aquasil LV and 0.79 MPa for Permadyne Garant L to 1.2 MPa for Impregum Penta Soft L. The loss tangent at 40 seconds tandelta (t(0)) varied among medium and low viscosity materials, ranging from liquid-like behavior of 4.3 for Permadyne Garant and less than unity or significant solid-like behavior for Impregum Penta. All materials showed tandelta values less than unity at their setting times. The temperature studies revealed significant changes in the kinetics of setting, with the setting time decreasing more than 3-fold between 25 degrees and 37 degrees C for the Aquasil Deca and the Impregum Penta Soft H and more than 2-fold for the Impregum Penta. At room temperature of 25 degrees C, the storage modulus for Impregum Penta Soft H and Aquasil Deca was 1.1% and 37.5% lower than Impregum Penta, respectively. CONCLUSION: The development of the viscoelastic rheological properties with time for 3 medium- and 4 low-viscosity impression materials at 33 degrees C showed significant differences in the setting time and the magnitude of the storage modulus. Compared with Impregum Penta, the new Impregum Penta Soft H was 6% less stiff when set, compared with 44% lower stiffness for the medium viscosity addition silicone. Temperature studies between 25 degrees C and 37 degrees C revealed strong temperature sensitivity of the kinetics of setting and evolution with time of their rheological properties. PMID- 12886209 TI - A simple method to estimate restoration volume as a possible predictor for tooth fracture. AB - STATEMENT OF PROBLEM: Many dentists cite the fracture risk posed by a large existing restoration as a primary reason for their decision to place a full coverage restoration. However, there is poor agreement among dentists as to when restoration placement is necessary because of the inability to make objective measurements of restoration size. PURPOSE: The purpose of this study was to compare a new method to estimate restoration volumes in posterior teeth with analytically determined volumes. MATERIAL AND METHODS: True restoration volume proportion (RVP) was determined for 96 melamine typodont teeth: 24 each of maxillary second premolar, mandibular second premolar, maxillary first molar, and mandibular first molar. Each group of 24 was subdivided into 3 groups to receive an O, MO, or MOD amalgam preparation design. Each preparation design was further subdivided into 4 groups of increasingly larger size. The density of amalgam used was calculated according to ANSI/ADA Specification 1. The teeth were weighed before and after restoration with amalgam. Restoration weight was calculated, and the density of amalgam was used to calculate restoration volume. A liquid pycnometer was used to calculate coronal volume after sectioning the anatomic crown from the root horizontally at the cementoenamel junction. True RVP was calculated by dividing restoration volume by coronal volume. An occlusal photograph and a bitewing radiograph were made of each restored tooth to provide 2 perpendicular views. Each image was digitized, and software was used to measure the percentage of the anatomic crown restored with amalgam. Estimated RVP was calculated by multiplying the percentage of the anatomic crown restored from the 2 views together. Pearson correlation coefficients were used to compare estimated RVP with true RVP. RESULTS: The Pearson correlation coefficient of true RVP with estimated RVP was 0.97 overall (P70%) in myeloid cell lines was observed for GGTI-286 (9/19), GGTI-298 (14/19), GGTI-2147 (16/19) and FTI L-744,832 (17/17). GGTI treatment of NB-4 cells resulted in an accumulation of cells in G(0)/G(1), whereas FTI L 744,832 primarily caused an increase in G(2)/M. FTI and GGTIs both induced apoptosis. In all cases, FTI/GGTI cotreatment led to synergistic cytotoxic effects in both myeloid cell lines (5/5) and primary AML cells (6/6). This synergy coincided with increased apoptosis. FTI/GGTI cotreatment caused an accumulation of unprocessed N-RAS and inactive N-RAS-RAF complexes. Our results suggest that alternative geranylgeranylation of N-RAS may represent an important mechanism of resistance to FTI monotherapy in myeloid leukemia cells. PMID- 12886238 TI - Risk-adapted postremission therapy in acute myeloid leukemia: results of the German multicenter AML HD93 treatment trial. AB - The objective of the AML HD93 treatment trial was to evaluate the outcome in young adults with acute myeloid leukemia (AML) after postremission therapy was stratified according to cytogenetically defined risk. The rationales for the study design were based (i) on previous favorable results with high-dose cytarabine in AML with t(8;21), inv/t(16q22) and in AML with normal karyotype, and ii) on encouraging results obtained in several phase II trials using autologous stem cell transplantation (SCT). Between July 1993 and January 1998, 223 eligible patients, 16-60 years of age with newly diagnosed AML other than French-American-British type M3/M3v, were entered into the trial. Risk groups were defined as follows: low risk: t(8;21) or inv/t(16q22); intermediate risk: normal karyotype; high risk: all other chromosomal abnormalities. Following intensive double induction therapy with idarubicin, cytarabine and etoposide, all patients in complete remission (CR) received a first consolidation therapy with high-dose cytarabine and mitoxantrone (HAM). A second consolidation therapy was stratified according to the risk group: low risk: HAM; intermediate risk: related allogeneic SCT or sequential HAM; high risk: related allogeneic or autologous SCT. Double induction therapy resulted in a high CR rate of 74.5%, and 90% of the responding patients were eligible for consolidation therapy. Survival for all 223 trial entrants was 40%, and for the 166 patients who entered CR, disease-free (DFS) and overall survival were 40 and 51% after 5 years, respectively. Within the low-, intermediate- and high-risk groups, DFS and survival after 5 years were 62.5 and 87, 40 and 49 and 17 and 26% respectively, without advantage for allogeneic transplantation in the intermediate- and high-risk groups. Postremission therapy-related mortality was 0, 7 and 14%, respectively. This study demonstrates the feasibility of cytogenetically defined risk-adapted consolidation therapy. The overall trial results are at least equivalent to those of published trials supporting the risk-adapted treatment strategy. PMID- 12886239 TI - Randomized comparison of interferon alpha and hydroxyurea with hydroxyurea monotherapy in chronic myeloid leukemia (CML-study II): prolongation of survival by the combination of interferon alpha and hydroxyurea. AB - The optimum treatment conditions of interferon (IFN) alpha therapy in chronic myeloid leukemia (CML) are still controversial. To evaluate the role of hydroxyurea (HU) for the outcome of IFN therapy, we conducted a randomized trial to compare the combination of IFN and HU vs HU monotherapy (CML-study II). From February 1991 to December 1994, 376 patients with newly diagnosed CML in chronic phase were randomized. In all, 340 patients were Ph/BCR-ABL positive and evaluable. Randomization was unbalanced 1:2 in favor of the combination therapy, since study conditions were identical to the previous CML-study I and it had been planned in advance to add the HU patients of study I (n=194) to the HU control group. Therefore, a total of 534 patients were evaluable (226 patients with IFN/HU and 308 patients with HU). Analyses were according to intention-to-treat. Median observation time of nontransplanted living patients was 7.6 years (7.9 years for IFN/HU and 7.3 years for HU). The risk profile (new CML score) was available for 532 patients: 200 patients (38%) were low, 239 patients (45%) intermediate, and 93 patients (17%) high risk. Complete hematologic response rates were higher in IFN/HU-treated patients (59 vs 32%). Of 169 evaluable IFN/HU treated patients (75%), 104 patients (62%) achieved a cytogenetic response that was complete in 12% (n=21), major in 14% (n=24), and at least minimal in 35% (n=59). Of the 534 patients, 105 (20%) underwent allogeneic stem cell transplantation in first chronic phase. In the low-risk group, 65 of 200 patients were transplanted (33%), 30 (13%) in the intermediate-risk group, and nine (10%) in the high-risk group. Duration of chronic phase was 55 months for IFN/HU and 41 months for HU (P<0.0001). Median survival was 64 months for IFN/HU and 53 months for HU-treated patients (P=0.0063). We conclude that IFN in combination with HU achieves a significant long-term survival advantage over HU monotherapy. In view of the data of CML-study I, these results suggest that IFN/HU is also superior to IFN alone. HU should be combined with IFN in IFN-based therapies and for comparisons with new therapies. PMID- 12886240 TI - The percentage of myeloperoxidase-positive blast cells is a strong independent prognostic factor in acute myeloid leukemia, even in the patients with normal karyotype. AB - To examine whether the percentage of myeloperoxidase (MPO)-positive blast cells is useful as a prognostic factor for acute myeloid leukemia (AML), cytochemical analysis of MPO was performed in 491 patients who were registered to the Japan Adult Leukemia Study Group-AML92 study. Patients were divided into two using the percentage of MPO-positive blast (high [>or=50%] and low (<50%)). Complete remission rates were 85.4% in the former and 64.1% in the latter (P=0.001). The overall survival (OS) and the disease-free survival (DFS) were significantly better in the high MPO group (48.3 vs 18.7% for OS, and 36.3 vs 20.1% for DFS, P<0.001, respectively). Multivariate analysis showed that both karyotype and the percentage of MPO-positive blast cells were equally important prognostic factors. The high MPO group still showed a better survival even when restricted to the intermediate chromosomal risk group or the patients with normal karyotype (P<0.001). The OS of patients with normal karyotype in the high MPO group was almost equal with that of the favorable chromosomal risk group. The percentage of MPO-positive blast cells is a simple and highly significant prognostic factor for AML patients, and especially useful to stratify patients with normal karyotype. PMID- 12886241 TI - Low expression of the myeloid differentiation antigen CD65s, a feature of poorly differentiated AML in older adults: study of 711 patients enrolled in ECOG trials. AB - CD65s appears when the progenitor antigen CD34 disappears, suggesting that this sialylated carbohydrate antigen marks a turning point in normal myeloid differentiation. We characterized acute myeloid leukemia (AML) with low CD65s expression (CD65s(low) AML) in 711 patients entered on seven Eastern Cooperative Oncology Group AML treatment trials (1986-1999). Of those, 198 (28%) qualified as having CD65s(low) AML. Morphologically, CD65s(low) AML was more common in FAB subgroups with minimal differentiation, M0/M1 (P=<0.0001). Early precursor antigens CD34, CD117 and terminal transferase were more frequent in CD65s(low) than CD65s(high) AML (P=<0.0001). Myeloperoxidase was present in fewer CD65s(low) myeloblasts, and the more mature myeloid antigens, CD15 and CD11b, were rarely detected (P=<0.0001). Yet, the two diagnoses did not differ in the distribution of cytogenetic prognostic groups or the occurrence of the multidrug-resistance mediator, P-glycoprotein. CD65s(low) AML patients were significantly older than CD65s(high) cases (P<0.0001). Furthermore, the incidence of CD65s(low) cases increased with age, from 20% in patients under the age of 50 years to 67% in patients older than 80 years (P<0.0001). Overall, complete remission (CR) rate and overall survival were comparable in CD65s(low) and CD65s(high) AML. However, among patients >55 years of age, CD65s(low) AML had a decreased CR rate of 33 vs 44% in CD65s(high) AML (P=0.055). Thus, CD65s(low) AML represents immunophenotypically undifferentiated disease and occurs predominantly in older adults. Although not statistically significant, the observed association between low CD65s expression and decreased CR rate only in patients over the age of 55 is intriguing. PMID- 12886242 TI - Expression of heat-shock protein 90 in glucocorticoid-sensitive and -resistant childhood acute lymphoblastic leukaemia. AB - Early reduction of leukaemic cells by chemotherapy is a strong predictor for treatment outcome in childhood acute lymphoblastic leukaemia (ALL). In ALL (Berlin-Frankfurt-Munster) trials, early treatment response is assessed by the in vivo response to glucocorticoids (prednisone response, PR), the molecular background of which is unknown. The intracellular effects of glucocorticoids (GCs) are mediated by the glucocorticoid receptor (GR). In the absence of GC, the inactive GR resides within a multiprotein complex, consisting predominantly of the chaperone protein hsp90 (heat-shock protein 90). Until now, studies targeting GC resistance mainly focused on GR disorders and alterations of genes known to be associated with drug resistance. In addition, the GR multiprotein complex was associated with GC resistance in in vitro studies. We performed a case-control study for PR to investigate the association of in vivo GC resistance and hsp90 expression in childhood ALL. Hsp90 expression was assessed using a real-time PCR approach (Taqman technology) and Western blot technology. In this setting, we found no association of in vivo GC resistance and hsp90 expression. Therefore, we conclude that the expression of hsp90, the major component of the GR activating complex, is of minor importance for the in vivo GC resistance in childhood ALL. PMID- 12886243 TI - Computational analysis of flow-cytometry antigen expression profiles in childhood acute lymphoblastic leukemia: an MLL/AF4 identification. AB - Precursor B-acute lymphoblastic leukemia (pB-ALL) is a heterogeneous disease and multiparameter flow cytometry, molecular genetics, and cytogenetic studies have all contributed to classification of subgroups with prognostic significance. Recently, gene expression microarray technology has been used to investigate lymphoblastic leukemias, demonstrating that known and novel pB-ALL subclasses can be separated on the basis of gene expression profiles. The strength of microarray technique lays in part in the multivariate nature of the expression data. We propose a parallel multiparametric approach based on immunophenotypic flow cytometry expression data for the analysis of leukemia patients. Specifically, we tested the potential of this approach on a data set of 145 samples of pediatric pB-ALL that included 46 samples positive for mixed lineage leukemia (MLL) translocations (MLL+) and 99 control pB-ALLs, negative for this translocation (MLL-). The expression levels of 16 marker proteins have been monitored by four color flow cytometry using a standardized diagnostic panel of antibodies. The protein expression database has been then analyzed using those univariate and multivariate computational techniques normally applied to mine and model large microarray data sets. Marker protein expression profiling not only allowed separating pB-ALL cases with an MLL rearrangement from other ALLs, but also demonstrates that MLL+ leukemias constitute a heterogeneous group in which MLL/AF4 leukemias represent a homogenous subclass described by a specific expression fingerprint. PMID- 12886244 TI - Minimal residual disease detection in childhood precursor-B-cell acute lymphoblastic leukemia: relation to other risk factors. A Children's Oncology Group study. AB - Minimal residual disease (MRD) can be detected in the marrows of children undergoing chemotherapy either by flow cytometry or polymerase chain reaction. In this study, we used four-color flow cytometry to detect MRD in 1016 children undergoing therapy on Children's Oncology Group therapeutic protocols for precursor-B-cell ALL. Compliance was excellent, with follow-up samples received at the end of induction on nearly 95% of cases; sensitivity of detection at this time point was at least 1/10,000 in more than 90% of cases. Overall, 28.6% of patients had detectable MRD at the end of induction. Patients with M3 marrows at day 8 were much more likely to be MRD positive (MRD+) than those with M2 or M1 marrows. Different genetically defined groups of patients varied in their prevalence of MRD. Specifically, almost all patients with BCR-ABL had high levels of end-of-induction MRD. Only 8.4% of patients with TEL-AML1 were MRD+>0.01% compared with 20.3% of patients with trisomies of chromosomes 4 and 10. Our results show that MRD correlates with conventional measures of slow early response. However, the high frequency of MRD positivity in favorable trisomy patients suggests that the clinical significance of MRD positivity at the end of induction may not be the same in all patient groups. PMID- 12886245 TI - Clonality profile in relapsed precursor-B-ALL children by GeneScan and sequencing analyses. Consequences on minimal residual disease monitoring. AB - Detection of minimal residual disease (MRD), using immunoglobulin (Ig) and T-cell receptor (TCR) gene rearrangements as clone-specific targets, represents the most recent development in diagnosis and treatment of acute lymphoblastic leukaemia (ALL). Nevertheless, risk of false-negative results, due to secondary or ongoing rearrangements of Ig/TCR genes during the disease course, might hamper MRD detection. Therefore, to gain extensive information on clonal stability, we performed PCR-GeneScan analysis of Ig/TCR gene rearrangements at diagnosis and subsequent relapse in bone marrow samples from 53 childhood precursor-B-ALL patients. In addition, sequencing analysis of junctional regions at diagnosis and relapse provided a detailed insight in the stability and changes of Ig/TCR gene rearrangements during the disease course. At least one stable clonal Ig/TCR target was found in 94% of patients. In three patients complete differences in Ig/TCR rearrangements between diagnosis and relapse were observed, suggesting relapse with a new clone. At relapse, 71% of diagnostic clonal PCR targets was conserved. Since the comparison of Ig/TCR gene rearrangements at diagnosis and relapse in our precursor-B-ALL patients did not show significant difference in the stability of different clonal PCR targets (IGH, 70%; IGK, 71%; TCRD, 67%; TCRG, 75%), we conclude that there is no 'preferential' clone-specific target for MRD monitoring. PMID- 12886246 TI - Evaluation of immunologic crossreaction of antiasparaginase antibodies in acute lymphoblastic leukemia (ALL) and lymphoma patients. AB - To evaluate how well antibodies to one asparaginase preparation predict or correlate with antibodies to another preparation in acute lymphoblastic leukemia (ALL) and lymphoma patients who did and did not have hypersensitivity reactions during chemotherapy. In all, 24 children with newly diagnosed ALL or lymphoma, who received Escherichia coli asparaginase 10 000 IU/m(2) IM thrice weekly for nine doses as part of multiagent induction and reinduction chemotherapy, and seven monthly doses during the first 7 months of continuation treatment, were studied. Plasma samples were collected at postinduction and at postreinduction. Six of 24 patients had no overt clinical reactions (nonreacting) and received only the E. coli preparation. Of these, 18 patients who had allergic reactions were switched to Erwinia asparaginase. A total of 18 patients had an anaphylactoid reaction to Erwinia asparaginase and were switched to receive polyethylene glycol (PEG) asparaginase. Antibody levels were measured by enzyme linked immunoadsorbent assay against all the three asparaginase preparations. At postinduction, antibodies against E. coli were higher in reacting patients (0.063+/-0.066) than in nonreacting patients (0.019+/-0.013) (P=0.03). At postreinduction, anti-Erwinia antibodies were significantly higher in reacting patients (0.431+/-0.727) than in nonreacting patients (0.018+/-0.009) (P=0.007). Anti-E. coli antibodies correlated with anti-PEG antibodies at postinduction (r=0.714, P<0.001) and at postreinduction (r=0.914, P<0.001), but did not correlate with anti-Erwinia antibodies at postinduction (r=0.119, P=0.580) and at postreinduction (r=0.078, P=0.716). The results indicate a crossreactivity between patient antibodies raised against natural E. coli and PEG asparaginase but not Erwinia asparaginase. PMID- 12886247 TI - Expression of the human homologue of rat NG2 in adult acute lymphoblastic leukemia: close association with MLL rearrangement and a CD10(-)/CD24( )/CD65s(+)/CD15(+) B-cell phenotype. AB - The expression of the chondroitin sulfate proteoglycan neuron-glial antigen 2 (NG2) has been demonstrated in association with rearrangement of the mixed lineage leukemia (MLL) gene in acute leukemia, but the frequency of NG2 expression in adult acute lymphoblastic leukemia (ALL) is yet unknown. We evaluated NG2 expression in 313 adult ALL patients by flow cytometry and simultaneously determined MLL rearrangement in 120 adult patients out of them with B-precursor ALL by reverse transcription-polymerase chain reaction and fluorescence in situ hybridization. A total of 57% of pro-B ALL, 2% of common ALL and 20% of pre-B ALL were NG2 positive, but NG2 was absent in T-ALL and mature B ALL. In B-precursor ALL, NG2 expression was significantly associated with a CD10( )/CD34(-)/CD24(-)/CD65s(+)/CD15(+)/CD13(-)/CD33(-) phenotype and showed a sensitivity, specificity and positive predictive value of 0.89, 0.89, and 0.93 for MLL rearrangement, respectively. NG2 was positive in three patients without detectable MLL rearrangement and negative in eight patients with MLL-AF4 transcripts. However, NG2 predicted with a 100% accuracy MLL rearrangement among patients disclosing a CD65s(+) and/or CD15(+) immunophenotype. In summary, NG2 adds to a more precise identification of high-risk adult ALL and should therefore be included into diagnostic marker panels. As NG2 is negative in non-malignant hematopoietic cells, this novel antigen might also serve in future studies as a powerful marker in monitoring minimal residual disease. PMID- 12886248 TI - Risk assessment in adult acute lymphoblastic leukaemia before early haemopoietic stem cell transplantation with a geno-identical donor: an easy clinical prognostic score to identify patients who benefit most from allogeneic haemopoietic stem cell transplantation. AB - In 1402 patients allografted in Europe during the period 1990-2000 with an HLA identical sibling in first remission (CR1), the median interval from CR1 to allotransplant (96 days) was a major prognostic factor, patients transplanted earlier having a worse outcome. We studied in depth the 414 fully evaluable patients transplanted less than 96 days after achieving CR1; in these patients, only three factors predicted for the outcome by multivariate analysis: patient age, CR1 achievement with one or more induction courses and the recipient/donor sex combination. These three factors overcame the information from cytogenetics and source of stem cells. Three prognostic groups could be identified in relation to the outcome, using a prognostic score affecting 1 to each poor risk factor (total from 0 to 3): Group 1 (good prognosis) includes patients <35 years old, achieving CR1 with one induction course and to be transplanted with any other sex combination than female to male (score 0); group 2 (intermediate) with one adverse factor (score 1); and group 3 (bad prognosis) with two or three adverse criteria (scores 2 and 3). In these three groups, the 3-year leukaemia-free survival was 56+/-5%, 48+/-4% and 29+/-4% and the overall survival was 65+/-5, 53+/-4 and 29+/-5%, respectively. Therefore, adult patients with ALL and a score of 0 or 1 are good candidates for an early transplant if they have an identical sibling donor. Patient age, response to induction and the sex of the HLA identical family donor (if existing) are the strongest easy predictors of the outcome for an early transplant in an adult patient with ALL. No additional information is mandatory. PMID- 12886249 TI - Myelodysplastic syndrome after acute promyelocytic leukemia: the European APL group experience. AB - With improved treatment of acute promyelocytic leukemia (APL) by all trans retinoic acid (ATRA) combined to anthracycline-aracytin chemotherapy (CT), a larger number of those patients may be at risk of late complications. Recently, the Rome group reported five cases of myelodysplastic syndrome (MDS) or acute myeloid leukemia (AML, non-APL) occurring during the course of 77 APL patients (6.5%) in complete remission (CR). From 1991 to 1998, we treated 677 newly diagnosed cases of APL, and 617 of them achieved CR with ATRA combined to CT (n=579) or CT alone (n=38); 246 of them received subsequent maintenance CT with 6 mercaptopurine and methotrexate. With a median follow-up of 51 months, 6 patients (0.97%) developed MDS, 13-74 months after the diagnosis of APL. In all six cases, t(15;17) and PML-RARalpha rearrangement were absent at the time of MDS diagnosis, and karyotype mainly showed complex cytogenetic abnormalities involving chromosomes 5 and/or 7, typical of MDS observed after treatment with alkylating agents, although none of the six patients had received such agents for the treatment of APL. Our findings suggest that MDS can indeed be a long-term complication in APL, although probably at lower incidence than that previously reported. PMID- 12886250 TI - Proteomic analysis of the cell-surface membrane in chronic lymphocytic leukemia: identification of two novel proteins, BCNP1 and MIG2B. AB - B-cell-specific plasma-membrane proteins are potential targets for either small molecule or antibody-based therapies. We have sought to annotate proteins expressed at the cell surface membrane in patients with chronic lymphocytic leukemia (CLL) using plasma-membrane-based proteomic analysis to identify previously uncharacterized and potentially B-cell-specific proteins. Proteins from plasma-membrane fractions were separated on one-dimensional gels and trypsinized fractions subjected to high-throughput MALDI-TOF mass spectrometry. Using this method, many known B-cell surface antigens were detected, but also known proteins not previously described in this disease or in this cellular compartment, including cell surface receptors, membrane-associated enzymes and secreted proteins, and completely unknown proteins. To validate the method, we show that BLK, a B-cell-specific kinase, is located in the CLL-plasma-membrane fraction. We also describe two novel proteins (MIG2B and B-cell novel protein #1, BCNP1), which are expressed preferentially in B cells. MIG2B is in a highly conserved and defined gene family containing two plasma-membrane-binding ezrin/radixin/moesin domains and a pleckstrin homology domain; the Caenorhabditis elegans homolog (UNC-112) is a membrane-associated protein that colocalizes with integrin at cell-matrix adhesion complexes. BCNP1 is a completely unknown protein with three predicted transmembrane domains, with three alternatively spliced final exons. Proteomic analysis may thus define new potential therapeutic targets. PMID- 12886251 TI - Coculture and transplant of purified CD34(+)Lin(-) and CD34(-)Lin(-) cells reveals functional interaction between repopulating hematopoietic stem cells. AB - The human hematopoietic stem cell compartment is comprised of repopulating CD34(+) and CD34(-) cells. The interaction between these subsets with respect to their reconstitution capacity in vivo remains to be characterized. Here, lineage depleted (Lin(-)) human CD34(+) and CD34(-) hematopoietic cells were isolated from human male and female umbilical cord blood (CB) and transplanted into immune deficient NOD/SCID EMV(null) mice, thereby allowing the use of human and Y chromosome-specific DNA sequences to discriminate human reconstitution contributed by CD34(+) vs CD34(-) repopulating stem cells. Although cultured human CB CD34(-)Lin(-) cells transplanted alone possessed only minimal repopulating capacity, with 15% of mice achieving low levels of engraftment, transplantation of cocultured male CD34(-)Lin(-) cells with female CD34(+)Lin(-) cells demonstrated human repopulation with a contribution from CD34(-)Lin(-) derived progeny in 80% of the recipients. After coculture and transplantation, male CD34(-)Lin(-) cells gave rise to primitive CD34(+)CD38(-) cells isolated in vivo, which demonstrated clonogenic progenitor function into multiple lineages. Taken together, our study indicates that the presence of CD34(+)Lin(-) cells in coculture enhanced the low repopulating function of human CD34(-)Lin(-) cells in vivo. We propose that CD34(+)Lin and CD34(-)Lin cells represent phenotypically distinct, but related cell types that exhibit unique and previously unappreciated functional interaction. PMID- 12886252 TI - The presence and longevity of peripherally expanded donor-derived TCRalphabeta+ mature T lymphocyte clones after allogeneic bone marrow transplantation for adult myeloid leukemias. AB - There are two major pathways for T-cell regeneration after allogeneic bone marrow transplantation; thymus-dependent T-cell differentiation of T-cell progenitors, and peripheral expansion of mature T cells in the graft. In order to learn to what extent the peripheral expansion of donor-derived mature T lymphocytes contributes to reconstitution of the TCRalphabeta+ T-cell repertoire after allogeneic bone marrow transplantation for adult myeloid leukemias, we pursued the fate of donor-derived T-cell clones using the amino-acid sequences of the complementarity-determining region 3 (CDR3) of the TCR-beta chain as a clonal marker. Clonal expansion of TCRalphabeta+ T lymphocytes with specific TCRBV subfamilies was identified in donor blood. Identical T-cell clones were not found in blood from recipients before transplantation. The donor-derived T-cell clones were identified in the circulating blood from recipients a few months after allogeneic bone marrow transplantation, and they remained in the blood for 18 months after transplant in two recipients, and for 56 months in one. These results suggest that the peripheral expansion of mature T lymphocytes in the graft makes a significant contribution to post-transplant T-cell regeneration during the early period of transplantation in humans, and that mature T cells can survive in recipients for several years. Further investigation will be required to explore which antigens drive the expansion of T-cell clones in donors and recipients, and the mechanisms of maintaining homeostatic balance between the thymus-dependent pathway and the peripheral expansion of mature T cells in post transplant T-cell regeneration. PMID- 12886253 TI - Defibrotide reduces procoagulant activity and increases fibrinolytic properties of endothelial cells. AB - Defibrotide (DF), a polydeoxyribonucleotide with antithrombotic properties, has recently proven effective in patients with severe hepatic veno-occlusive disease (VOD), a life-threatening complication of high-dose chemo/radiotherapy regimens for stem cell transplantation. To understand the mechanism of its beneficial effect, we studied the impact of DF on the expression of tissue factor (TF) and fibrinolytic proteins (PAI-1 and t-PA) on endothelial cells. The in vitro response to DF of two types of human endothelial cells (ECs) of different origins, that is from macrovascular (HUVEC) and microvascular (HMEC-1 cell line) beds, was evaluated in the presence or absence of a proinflammatory stimulus (ie bacterial endotoxin, LPS). The results show that DF was able to significantly reduce the LPS-induced TF expression by HMEC-1, and less prominently by HUVEC. In addition, DF importantly influenced the fibrinolytic properties of both HMEC-1 and HUVEC. Specifically, it dose-dependently counteracted the LPS-induced increase in PAI-1 levels and decrease in t-PA activity expression. It also significantly incremented t-PA antigen in resting EC. Decreasing the procoagulant activity and increasing the fibrinolytic potential of EC favors an anticoagulant phenotype of the endothelium, which may protect from fibrin deposition and vascular occlusion. PMID- 12886254 TI - Tumor outgrowth in peripheral blood mononuclear cell-injected SCID mice is not associated with early Epstein-Barr virus reactivation. AB - Epstein-Barr virus (EBV)-positive B-cell lymphoproliferative disease develops in severe combined immunodeficient (SCID) mice inoculated with peripheral blood mononuclear cells (PBMC) from EBV(+) individuals (SCID/hu mice). In this study, we investigated the contribution of EBV reactivation and de novo infection of B lymphocytes to tumor outgrowth in SCID/hu mice. Evaluation of BZLF-1, an early EBV activation transcript, in cells recovered from the mouse peritoneal cavity within 16 days following PBMC transfer did not reveal EBV reactivation, while BZLF-1 expression was only detected in tumor masses or in vitro established lymphoblastoid cell lines. To confirm these data by a different strategy, we coinjected PBMC from seropositive donors with purified B cells from seronegative donors of different sex. Fluorescence in situ hydridization analysis of the resulting tumor masses disclosed that the overwhelming majority of lymphoma cells originated from the seropositive donor, implying that no substantial in vivo production and transmission of virus had occurred. Further, treatment of SCID/hu mice with ganciclovir did not prevent lymphoma development. Our results suggest that in the SCID/hu mouse, early EBV replication and secondary infection of bystander B cells does not occur, and that the direct outgrowth of the transformed B lymphocytes present within the PBMC inoculum is the predominant mechanism, which leads to lymphoma generation in this experimental model. PMID- 12886255 TI - Inactivation of the E3/LAPTm5 gene by chromosomal rearrangement and DNA methylation in human multiple myeloma. AB - Chromosomal band 1p34-36 is a commonly rearranged locus in many types of cancers. We cloned the breakpoint region of a chromosomal translocation, t(1;14)(p34;q32), found in the human multiple myeloma (MM) cell line, ODA. This rearrangement occurred between the nearby switch region of the immunoglobulin heavy chain (IgH) gene (Sgamma3) at 14q32 and the first intron of the human retinoic acid-inducible E3 protein (E3)/lysosome-associated protein, transmembrane-5 (LAPTm5) gene at the 1p34 locus. Consequently, the E3 gene, which is a hematopoietic cell-specific transcript induced by retinoic acid and located at the rearranged allele, was interrupted within its coding region and was not expressed in the ODA cell line in spite of the other allele still being intact. The expression derived from the remaining intact allele in ODA cells was silenced by DNA methylation at sequences within the first intron around a GC-rich EagI site. Interestingly, the silenced expression of E3 mRNA due to DNA methylation of intron 1 sequences was frequently encountered in MM cells [6/10 (60%) of MM cell lines tested], while E3 is expressed in normal plasma cells and in most other hematopoietic cell lines including those of B-cell lineage. Thus, as the E3 protein has been suggested to be involved in cellular differentiation and apoptotic pathways in certain cell types, our results suggest that loss of E3 gene expression might be a crucial event during the progression of human MM. PMID- 12886256 TI - Treatment of relapsed B-cell non-Hodgkin's lymphoma with a combination of chimeric anti-CD20 monoclonal antibodies (rituximab) and G-CSF: final report on safety and efficacy. AB - Antibody-dependent cellular cytotoxicity (ADCC) is one of the possible mechanisms of action of the chimeric CD20 monoclonal antibody IDEC-C2B8 (rituximab). As granulocyte-colony stimulating factor (G-CSF) greatly enhances the cytotoxicity of neutrophils in ADCC, the efficacy of rituximab might be enhanced by the addition of G-CSF. In a phase I/II clinical trial, we investigated the safety and efficacy of the combination of rituximab and G-CSF (5 microg/kg/day, administered for 3 days, starting 2 days before each infusion) in 26 relapsed low-grade lymphoma patients. Adverse events occurred in 25/26 patients and mainly consisted of (grade I/II) fever (29%) and allergic reactions (19%). In phases I and II (375 mg/m(2) rituximab+G-CSF), 19 patients were evaluable for efficacy. The response rate was 42% (8/19; 95% CI 20-67%), with 16% (3/19) complete remissions and 26% (5/19) partial remissions. The median duration of response was 18 months, the median time to progression was 24 months. We conclude that the combination of rituximab and G-CSF is well tolerated. Although the overall response rate seems comparable to that reported for rituximab monotherapy, remission duration in this pilot phase II study is remarkably long. Randomized comparison with rituximab monotherapy should substantiate this promising finding. PMID- 12886257 TI - Fusion of AML1/Runx1 to copine VIII, a novel member of the copine family, in an aggressive acute myelogenous leukemia with t(12;21) translocation. PMID- 12886258 TI - Cryptic rearrangement involving MLL and AF10 occurring in utero. PMID- 12886259 TI - Sequential combination of thalidomide and erythropoietin determines transfusion independence and disease control in idiopathic myelofibrosis previously insensitive to both drugs used as single agents. PMID- 12886260 TI - Monitoring all-trans-retinoic acid-induced differentiation of human acute promyelocytic leukemia NB4 cells by Fourier-transform infrared spectroscopy. PMID- 12886261 TI - Minimal residual disease in acute myeloid leukemia is predicted by P-glycoprotein activity but not by multidrug resistance protein activity at diagnosis. PMID- 12886262 TI - Absence of somatic mutations within the Runt domain of AML2/RUNX3 in acute myeloid leukaemia. PMID- 12886263 TI - SOCS-1 gene methylation is frequent but does not appear to have prognostic value in patients with multiple myeloma. PMID- 12886264 TI - Amplification of band q22 of chromosome 21, including AML1, in older children with acute lymphoblastic leukemia: an emerging molecular cytogenetic subgroup. PMID- 12886265 TI - Classification of mature T-cell leukemias. PMID- 12886267 TI - Identification of CD14 as a predictor for leukemic dendritic cell differentiation in acute myeloid leukemia. PMID- 12886270 TI - Plant polyploidy: gene expression and genetic redundancy. PMID- 12886275 TI - Analogous pleiotropic effects of insecticide resistance genotypes in peach-potato aphids and houseflies. AB - We show that single-point mutations conferring target-site resistance (kdr) to pyrethroids and DDT in aphids and houseflies, and gene amplification conferring metabolic resistance (carboxylesterase) to organophosphates and carbamates in aphids, can have deleterious pleiotropic effects on fitness. Behavioural studies on peach-potato aphids showed that a reduced response to alarm pheromone was associated with both gene amplification and the kdr target-site mutation. In this species, gene amplification was also associated with a decreased propensity to move from senescing leaves to fresh leaves at low temperature. Housefly genotypes possessing the identical kdr mutation were also shown to exhibit behavioural differences in comparison with susceptible insects. In this species, resistant individuals showed no positional preference along a temperature gradient while susceptible genotypes exhibited a strong preference for warmer temperatures. PMID- 12886276 TI - Effective size in simple metapopulation models. AB - A coalescent argument is used to derive the effective size in simple models with recurrent local extinctions. Several alternative methods of derivation of this result are given and compared to earlier analyses of this problem. The different methods described in this paper all give the same result, which differs from earlier ones. For two published sets of estimates of demographic parameters, metapopulation structure appears to result in a moderate reduction of effective size relative to total adult population size. PMID- 12886277 TI - Family and population effects on disease resistance in a reptile. AB - Despite its importance in evolutionary biology, studies of the pattern of disease resistance in natural populations are rare. In this paper, we report patterns of infection of a viral eye disease in juvenile Swedish common lizards (Lacerta vivipara). Females were sampled at random from natural populations immediately prior to parturition with equal exposure of pathogens for all lizards once in captivity. No causative agents could be found that linked risk of disease to maternal/interfollicular transfer of pathogens. The results show that a major factor influencing offspring susceptibility is family identity, suggesting heritable variation in pathogen resistance. Our interpopulation comparison provides additional support for a link between genetics and disease resistance. Lizards in northern Sweden were not only more susceptible to the disease but were also more health compromised once infected, with relatively more reduced growth rate and increased mortality than lizards from the south. This scenario suggests that southern lizards have been under selection for resistance to this pathogen, whereas northern lizards have not, or at least not to the same degree. Thus, this study confirms the importance of genetic (family) effects on pathogen resistance with variation in this trait among natural populations. PMID- 12886278 TI - Genetic and maternal effect influences on viability of common frog tadpoles under different environmental conditions. AB - The influence of environmental stress on the expression of genetic and maternal effects on the viability traits has seldom been assessed in wild vertebrates. We have estimated genetic and maternal effects on the viability (viz probability of survival, probability of being deformed, and body size and shape) of common frog, Rana temporaria, tadpoles under stressful (low pH) and nonstressful (neutral pH) environmental conditions. A Bayesian analysis using generalized linear mixed models was applied to data from a factorial laboratory experiment. The expression of additive genetic variance was independent of pH treatments, and all traits were significantly heritable (survival: h2 approximately 0.08; deformities: h2 approximately 0.26; body size: h2 approximately 0.12; body shape: h2 approximately 0.14). Likewise, nonadditive genetic contributions to variation in all traits were significant, independent of pH treatments and typically of magnitude similar to the additive genetic effects. Maternal effects were large for all traits, especially for viability itself, and their expression was partly dependent on the environment. In the case of body size, the maternal effects were mediated largely through egg size. In general, the results give little evidence for the conjecture that environmental stress created by low pH would impact strongly on the genetic architecture of fitness-related traits in frogs, and hamper adaptation to stress caused by acidification. The low heritabilities and high dominance contributions conform to the pattern typical for traits subject to relatively strong directional selection. PMID- 12886279 TI - Spatial patterns of mitochondrial and nuclear gene pools in chamois (Rupicapra r. rupicapra) from the Eastern Alps. AB - We have assessed the variability of maternally (mtDNA) and biparentally (allozymes) inherited genes of 443 chamois (Rupicapra r. rupicapra) from 19 regional samples in the Eastern Alps, to estimate the degree and patterns of spatial gene pool differentiation, and their possible causes. Based on a total mtDNA-RFLP approach with 16 hexanucleotide-recognizing restriction endonucleases, we found marked substructuring of the maternal gene pool into four phylogeographic groups. A hierarchical AMOVA revealed that 67.09% of the variance was partitioned among these four mtDNA-phylogroups, whereas only 8.04% were because of partitioning among regional samples within the populations, and 24.86% due to partitioning among individuals within regional samples. We interpreted this spatial pattern of mtDNA variability as a result of immigration of chamois from different Pleistocene refugia surrounding the Alps after the withdrawal of glaciers, rather than from topographic barriers to gene flow, such as Alpine valleys, extended glaciers or woodlands. However, this striking geographical structuring of the maternal genome was not paralleled by allelic variation at 33 allozyme loci, which were used as nuclear DNA markers. Wright's hierarchical F statistics revealed that only < or =0.45% of the explained allozymic diversity was because of partitioning among the four mtDNA-phylogroups. We conclude that this discordance of spatial patterns of nuclear and mtDNA gene pools results from a phylogeographic background and sex-specific dispersal, with higher levels of philopatry in females. PMID- 12886280 TI - A linkage map for the hybridising toads Bombina bombina and B. variegata (Anura: Discoglossidae). AB - Stable hybrid zones in which ecologically divergent taxa give rise to a range of recombinants are natural laboratories in which the genetic basis of adaptation and reproductive isolation can be unraveled. One such hybrid zone is formed by the fire-bellied toads Bombina bombina and B. variegata (Anura: Discoglossidae). Adaptations to permanent and ephemeral breeding habitats, respectively, have shaped numerous phenotypic differences between the taxa. All of these are, in principle, candidates for a genetic dissection via QTL mapping. We present here a linkage map of 28 codominant and 10 dominant markers in the Bombina genome. In an F2 cross, markers that were mainly microsatellites, SSCPs or allozymes were mapped to 20 linkage groups. Among the 40 isolated CA microsatellites, we noted a preponderance of compound and frequently interleaved CA-TA repeats as well as a striking polarity at the 5' end of the repeats. PMID- 12886281 TI - Spatial and temporal genetic variability in French populations of the peach potato aphid, Myzus persicae. AB - The peach-potato aphid, Myzus persicae (Sulzer), has a complex reproductive mode in which some lineages reproduce by continuous parthenogenesis, whereas others reproduce sexually once a year. The climate is thought to act directly on the reproductive mode, because sexual eggs are the only form that can resist frost in cold regions. Sexual reproduction necessitates an obligatory host alternation that may result in long-distance dispersal. Here, we examined the genetic variability at seven microsatellite loci of populations of M. persicae in France, where both reproductive modes occur. We provide clear genetic evidence that the breeding system affects genotypic variability, as cyclically parthenogenetic aphids are far more variable than their obligately parthenogenetic counterparts. A temporal decrease in genetic variability and a temporal genetic differentiation effect suggest the existence of selective factors that play an important role in shaping the genetic structure of M. persicae populations. Lastly, differences in the population structure between reproductive modes suggest that the migration associated with the change of host during sexual reproduction lowers the level of population differentiation. PMID- 12886282 TI - Intraspecific variation, sex-biased dispersal and phylogeography of the eastern grey kangaroo (Macropus giganteus). AB - Genetic information has played an important role in the development of management units by focusing attention on the evolutionary properties and genetics of populations. Wildlife authorities cannot hope to manage species effectively without knowledge of geographical boundaries and demic structure. The present investigation provides an analysis of mitochondrial DNA and microsatellite data, which is used to infer both historical and contemporary patterns of population structuring and dispersal in the eastern grey kangaroo (Macropus giganteus) in Australia. The average level of genetic variation across sample locations was one of the highest observed for marsupials (h=0.95, HE=0.82). Contrary to ecological studies, both genic and genotypic analyses reveal weak genetic structure of populations, where high levels of dispersal may be inferred up to 230 km. The movement of individuals was predominantly male-biased (average Nem=22.61, average Nfm=2.73). However, neither sex showed significant isolation by distance. On a continental scale, there was strong genetic differentiation and phylogeographic distinction between southern (TAS, VIC and NSW) and northern (QLD) populations, indicating a current and/or historical restriction of gene flow. In addition, it is evident that northern populations are historically more recent, and were derived from a small number of southern founders. Phylogenetic comparisons between M. g. giganteus and M. g. tasmaniensis indicated that the current taxonomic status of these subspecies should be revised as there was a lack of genetic differentiation between the populations sampled. PMID- 12886283 TI - Ribosomal DNA variation, recombination and inheritance in the basidiomycete Trichaptum abietinum: implications for reticulate evolution. AB - Two divergent nuclear ribosomal DNA (nrDNA) types, designated alpha and beta, were found distributed in 11 North European populations of the basidiomycete Trichaptum abietinum. These types differed by a 220 bp indel in the internal transcribed spacer 1 (ITS1) sequence and a number of linked substitutions and small indel motives in the internal transcribed and intergenic spacers (ITS1, ITS2, IGS1 and IGS2). The alpha and beta haplotypes co-occurred in heterozygous somatic individuals (dikaryons) and segregated in a Mendelian fashion in monokaryotic single spore progenies. This result suggests that the haplotypes are encoded in different nuclei of field-collected dikaryons and inherited as a single locus. No meiotic recombinants were observed among the sequenced monokaryons. Population genetic analyses by PCR-RFLP revealed that a low frequency of evolutionary intermediate nrDNA types also existed in natural populations, presumably as a result of meiotic recombination of alpha and beta nrDNA. The existence of divergent nrDNA types in T. abietinum could be a result of a former independent evolution followed by a hybridization event. Phylogenetic analyses of ITS sequences suggest that the sister taxon T. fusco-violaceum has been involved in the evolutionary history of T. abietinum. Sequence polymorphisms observed in the translation elongation factor 1alpha (efa) and glyceraldehyde-3 phosphate dehydrogenase (gpd) genes, did not reveal two well-defined types of these genes. The results are discussed in the light of other evolutionary mechanisms as well. PMID- 12886284 TI - On methods of spatial analysis for genotyped individuals. AB - Spatial autocorrelation methods have commonly been applied to individual-based spatial genetic studies, although their properties and the relations among the statistics have not been carefully examined. This paper first introduces a reformulation of widely used spatial statistics using point processes. When Moran's I statistics are applied to allele frequencies within an individual, the frequencies are no longer continuous variables but have only three discrete values and specific interpretations of Moran's I statistics and the number of alleles in common (NAC) can be expressed as the weighted sum of join-count statistics. The distributions of minor genotypes are amplified in Moran's I depending on the allele frequency in the population, while NAC uses a constant weighting system. Under the point process framework, spatial analysis can be conducted on the common theoretical base, from individual locations to genetic distributions of different levels, (for example, genotype and allele). The methodology is demonstrated by application to field data for molecular ecological studies of Fagus crenata population dynamics. PMID- 12886285 TI - Nuclear and chloroplast genetic structure indicate fine-scale spatial dynamics in a neotropical tree population. AB - Dicorynia guianensis is a canopy tree, endemic to the tropical rain forest of French Guiana. We compared generational and spatial genetic structure for maternally and biparentally inherited markers in two cohorts (adult and seedling) in order to infer processes shaping the distribution of genetic diversity. The study was conducted on a 40 ha study plot located at Paracou near Kourou, where 172 adults trees and 375 saplings were sampled. Aggregation of trees was therefore suggested at different distances, ranging from 100 to 400 m. There was a strong link between demographic and genetic spatial structures at small distances (less than 100 m) that is likely to be the consequence of restricted seed dispersal. Genetic differentiation was more pronounced between spatial aggregates than between cohorts. Despite the spatial differentiation, the species was able to maintain high levels of diversity for maternal genomes, suggesting rapid turnover of aggregates. Spatial autocorrelation was larger for chloroplast than nuclear markers indicating a strong asymmetry between pollen and seed flow. Fixation indices indicated a lower heterozygote deficiency for the adults, maybe because of gradual elimination of selfed trees. Genetic relatedness at lower distances was higher in adult trees than in saplings, as a result of generation overlapping in the adult cohort. Overall, our results confirm earlier biological knowledge about the dispersion mechanisms of the species, and lead to an enhanced role of spatial processes in the dynamics of genetic diversity of D. guianensis. PMID- 12886286 TI - Coordinates and peer review. PMID- 12886287 TI - The long and short of adaptor appendages. PMID- 12886288 TI - Now, swing your partner! 3D-domain switching of WASP activates Arp2/3 complex. PMID- 12886289 TI - The IgA receptor complex: a two-for-one deal. PMID- 12886290 TI - P450 on drugs. PMID- 12886295 TI - [The future of registers and epidemiological surveillance systems]. PMID- 12886291 TI - Pulling the calcium trigger. PMID- 12886296 TI - [Liver transplantation in year 2000]. PMID- 12886297 TI - [Aetiology and bacterial resistance in urinary infections of outpatients]. AB - BACKGROUND: Knowledge of the etiological agents of infections of the urinary tract and of the map of the antimicrobial sensitivity of each geographic zone is a practice recommended for improving the use of antibiotics and for facilitating the choice of empirical treatment. The aim of this study is to provide up-to-date information on the pathogen flora of outpatients urinary infections in our health area and their sensitivity to antimicrobials, based on the urinary cultivations carried out in the Microbiology laboratory of the Solchaga Outpatient Surgery during the last four years. MATERIAL AND METHODS: Between 1996 and 1999, we carried out 92,816 urinary cultivations of outpatients. The urine specimens were sown on plaques with chocolate agar and MacKonkey agar with calibrated loop. The antibiogram was carried out using the Kirby-Bauer method. RESULTS: The percentage of positive urinary cultivations was 17%. Gram-negative bacilli represented 88.9% of the isolations, Gram-positive cocci 11% and yeasts 0.1%. Escherichia coli was the most frequently isolated germ (71%). The sensitivity studies show that amoxycillin-clavulanic acid, first generation cephalosporins and fosfomycin maintain a sensitivity above 80%. CONCLUSIONS: Up-to-date knowledge of the most common pathogen flora and their sensitivity to antimicrobials makes it possible for a more suitable selection to be made of the antibiotic treatment when the sensitivity pattern of the responsible germ is unavailable. In the light of the results we can conclude that amoxycillin-clavulanic acid, first generation cephalosporins and fosfomycin show a good activity in vitro, and should be considered as empirical treatments for infections of the urinary tract. PMID- 12886298 TI - [Measurement of stool composition in children with celiac disease using near infrared reflectance spectroscopy]. AB - Celiac disease is a disorder caused by gluten-sensitivity which, when manifested in its classical digestive form, frequently presents a malabsorption syndrome. The aim of this study is to evaluate the faecal composition in celiac children with malabsorption syndrome at the moment of diagnosis by using near-infrared reflectance spectroscopy and to compare it with that of healthy children. Thirty children with biopsy-proven celiac disease and 86 age-matched control children were recruited in our study. Children collected 24 hour faecal specimens and the analyses of faeces, water, fat, nitrogen and sugar were performed using near infrared reflectance spectroscopy. Results show that celiac children daily eliminate a significantly greater quantity of water, fat, nitrogen and sugar than those in the control group. This might be due to the higher weight of faeces eliminated in the celiac group and, of course, to their celiac condition. Of all the nutrients, faecal fat is the substance which undergoes the greatest change, indicating that this nutrient continues to be the best parameter for dealing with patients with malabsorption syndrome. Near-infrared reflectance spectroscopy appears to be a useful tool for assessing stool composition in celiac disease. PMID- 12886299 TI - [Multiple sclerosis and fatigue]. AB - Fatigue is a frequent symptom in multiple sclerosis, known since the first descriptions of the disease, and it has been the object of special interest in recent years due to the great influence it has on the quality of life of those suffering from the disease. The fatigue of multiple sclerosis has clinical characteristics that differentiate it from the fatigue of other systemic diseases. At the same time, it is highly variable between patients and a clear distinction can be made between three types: astenia, fatigability and the worsening of symptoms with exercise. The physiopathological bases are not fully established, but there is data indicating that astenia is related to immunoactivation and evidence of the relation of fatigability to alterations of the pyramidal tract. Another point of interest in the study of fatigue is how to objectivise and quantify it, as well as the neurophysiological techniques for its evaluation and the measurement scales most frequently employed. The paper also sets out the principal therapeutic tests and considers the relative efficacy of aminopyridine and amantadine in alleviating it. PMID- 12886300 TI - [Genetic alterations in hematological neoplasias of lymphoid origin: implications for clinical practice]. AB - The improvement of the conventional cytogenetic techniques, the development of molecular cytogenetics and the application of techniques of molecular biology to genetic analysis have led to an authentic revolution in the knowledge of the processes implied in the development and progression of lymphoid neoplasias. In this way, a great part of the alterations present in malign cells have been characterised, and the genes involved in the transformative process have been established. This has important consequences for the clinical handling of this type of disease and makes possible a more exact diagnosis through a systematisation of the different entities based on their biological characteristics. On the other hand, the introduction of new techniques of analysis, such as real time PCR, will make it possible to monitor the disease quantitatively, making it possible to evaluate response to the different treatments and to establish predictive values for relapses. In the future, all of this knowledge will make it possible to establish genotype-specific therapies and to develop new medicines aimed at the alteration responsible for the malignant process and with less undesired collateral effects. PMID- 12886301 TI - [Epidemiology of HIV and AIDS infection in Navarra]. AB - The incidence of AIDS has been declining in the European Union since 1996 as a result, at least in part, of the use of more efficient treatments for HIV infection. The same tendency can be observed in Navarra, where the average has dropped from 70 annual cases in the first five years of the 1990s to 30 cases in the year 1999. In Navarra, the most frequent category of transmission was the consumption of intravenous drugs, a practice referred to by 72% of those diagnosed with AIDS between 1985 and 1999. 21% were attributed to sexual transmission (7% to homosexual practices and 14% to heterosexual), 2.5% to other mechanisms (mother-child, hemoderivates, etc.) and in the remaining 4.6% information was unavailable on the probable mechanism of transmission. With respect to the incidence of HIV infection, between 1985 and December 1999, 2,379 cases were diagnosed in Navarra. The annual number of new diagnoses of HIV infection showed two maximum peaks in the years 1987 and 1991, with over 200 annual cases. A progressive decline can be observed from 1993 onwards, falling to 40 cases in 1999. The figure of over 2,000 cases of HIV infection contrasts with the 672 cases of AIDS registered in this autonomous community up until the year 2000, and resituates the epidemic in more certain terms. Although the cases of AIDS and HIV have fallen in Navarra in recent years, the data from the microbiology laboratories and the hospital services that treat persons with HIV infection indicate that a considerable number of new cases of infection will continue to occur, which justifies the need for maintaining the prevention programs. PMID- 12886302 TI - [Incidence of cancer in Navarra. 1995-96]. AB - The present paper studies the results of the incidence of cancer in Navarra during 1995-96. The collection of data for the years 1995-1996 was carried out in an active form in the information sources of the register. The procedures for collection, control of duplicates and processing of previous years was maintained. Both the quality indicators and the calculations of the different rates were obtained following the recommendations proposed by the International Agency for Research on Cancer (IARC). In 1995-96, 5,667 incident cases of malign tumours were registered in Navarra; 58% occurred in men. Excluding non-melanoma skin cancer, the gross rate of incidence was 513.7 in men and 344.9 in women, the rate adjusted to the world population was 301 per 100,000 for men and 192 per 100,000 for women. Fifty six per cent of all the cases of cancer diagnosed during 1995-1996 in men occurred in the locations of lung, prostate, colon, stomach and bladder. In women the locations of breast, colon, body of the uterus, stomach and rectum represented 51% of total cases. PMID- 12886303 TI - [Aspects of pre-operational information of the patient in emergency surgery]. AB - BASIS: Under the conditions in which hospital activity is carried out, information can present important shortcomings. The aim of our study is to analyse the perception held by the patients of the information provided before an emergency surgical operation. MATERIAL AND METHODS: The study was carried out by means of a survey to 115 patients (66 men and 49 women) before they received emergency surgery, in the Hospital de Navarra, in the months from March to December, 1999. The sample was selected when the researchers responsible for this work were on duty. The majority of the operations corresponded to general surgery (39.1%) and traumatology (42.6%). The average age was 45.7 years (SD = 21.57, range 15-85 years). RESULTS: The results express the relations and differences between the degree of information requested and the cultural level, age and sex of the patient. Eighty seven per cent of the patients had been informed about the operation they were about to undergo. Seventy eight per cent had not signed their consent for such an operation and the doctors signature appeared in only 6%. Some 8.7% did not feel they had received information from anybody. Seventy three per cent understood the information provided. The information was considered insufficient by 44.6% of the patients. Forty five per cent were able to express their doubts to the informant, and 96.4% of them were satisfied with the clarifications. CONCLUSIONS: The patients considered themselves to be informed about the surgical process, but very few of them about the most relevant sensations they were to experience in post-operation. Information should be supplied that includes both types of content. PMID- 12886304 TI - [Herpes zoster and post-infection sequels to maternal chicken pox. One case]. PMID- 12886306 TI - [Ethics and evidence based medicine]. PMID- 12886305 TI - [Abdominal pain and vasculitis. Case of Henoch-Schonlein purpura]. PMID- 12886307 TI - [Nitric oxide: synthesis, neuroprotection and neurotoxicity]. PMID- 12886308 TI - [The application of the Minimum Data Set (MDS) in the surveillance of nosocomial infections]. AB - BASIS: Analysis of the application of the Minimum Data Set in the vigilance of nosocomial infection. Study of the risk of nosocomial infection according to the Deyo-Charlson Comorbidity Index. METHODS: The database of the Minimum Data Set of the hospital was related with the database containing the infections collected by the Infectious Diseases Surveillance Unit of the Preventive Medicine Service for the year 1997. Surgical procedures were classified by the North American system of vigilance of nosocomial infections. The diagnoses appearing in the Deyo Charlson Index were localised. The rates of nosocomial infection were calculated in relation to the presence of comorbidities and procedures. RESULTS: Not all discharges were codified (6.2%). Mortality and the rate of nosocomial infection were lower in codified discharges than in non-codified (4 and 1.5 respectively, p<0.01). Amongst the codified discharges, the rates of nosocomial infection and the surgical site are higher amongst surgery patients than in non-surgery cases according to the North American vigilance system of nosocomial infections. Nosocomial infection is associated with a higher hospital mortality. As scoring increases on the Deyo-Charlson Index, there is a rise in the rate of nosocomial infection (1.8% to 9.9%), average stay (average 14-22 days) and hospital mortality (0.2% to 17.8%). CONCLUSIONS: The validity of these results depends on the validity of the data gathered in the Minimum Data Set, which is in its turn determined, amongst other factors, by the quality of the Discharge Report with respect to its inclusion of diagnoses and principal and secondary procedures, and by exhaustiveness in the codification of hospital discharges. PMID- 12886309 TI - [Value of the expression of p21RAS, P53, Bcl-2 oncoproteins and Ki-67(MIB-1) antigen of cellular proliferation in the diagnosis and prognosis of thyroi]. AB - INTRODUCTION: The thyroid gland is a good organ for the study of genetic alterations in tumoural development and progress. The study of oncogenes and antigens of cellular proliferation is of interest. MATERIAL AND METHODS: 36 cases of thyroid adenomas, 58 papillary carcinomas, 32 follicular carcinomas, 2 anaplastic carcinomas and 12 medullary carcinomas were selected. Clinical and histological prognostic factors were studied and an immunohistochemical study with Ki-67 (MIB-1), p53, Bcl-2 and p21 RAS antibodies was performed. RESULTS: The proliferative rate Ki-67 (MIB-1) showed statistical differences in adenomas and follicular carcinomas. The p53 protein did not appear in adenomas, but was shown in those carcinomas with bigger histological dedifferentiation. A relationship was also established with greater age, larger size and progression of the tumour. The Bcl-2 protein was high in the normal thyroid, in the adenomas and in the carcinomas, decreasing in correlation with histological dedifferentiation, and totally disappearing in the anaplastic carcinomas. In medullary carcinomas its disappearance was correlated with mortality. The intense expression of the p21 RAS protein in papillary carcinomas is statistically correlated with advanced clinical studies. CONCLUSIONS: The differences in the proliferative rate between adenomas and follicular carcinomas can be of help in differential diagnosis. The p53 manifestation and the loss of the Bcl-2 expression are correlated with the histological redifferentiation process. An intense manifestation of p21 RAS in papillary carcinoma is correlated with tumoural progression. PMID- 12886310 TI - [Outbreak of multiresistant tuberculosis]. AB - BASIS: Six cases of multiresistant tuberculosis diagnosed in the Hospital of Navarra in 1996 were studied. MATERIAL AND METHODS: Sensitivity tests were carried out using the BACTEC 460TB system against four main anti-tuberculosis medicines: Isoniazid, Streptomycin, Rifampizin and Ethambutol. "Fingerprinting" techniques by means of RFLP with IS 6110 were used in the epidemiological study. RESULTS: Four of the patients were also infected with HIV. All of them had a fatal short-term prognosis. These four showed resistance to the drugs under study; in three resistance was primary, and secondary in the fourth. The molecular study showed that three HIV+ patients had identical patterns of restriction fragment length polymorphism (RFLP). Two were brothers and nosocomial infection was demonstrated in the third. On the other hand, the RPLF pattern of these patients showed great similarity to that of one of the HIV- patients, which suggests a clone origin of the strains, no other epidemiological relationship being found. The RPLF pattern of the other HIV+ patient was identical to that of another of his strains, isolated 14 months before, with the first strain sensitive. The other two patients were HIV- women, and they evolved favourably. In one, who was diabetic, a multiresistant tuberculosis had been diagnosed 7 years before, remaining asymptomatic up until the present. The other involved a secondary resistance due to deficient treatment. CONCLUSIONS: The appearance of multiresistant tuberculosis is a very serious problem amongst HIV+ patients, especially at a time when their life expectancy has improved considerably due to the new therapies. In HIV- patients, multiresistant tuberculosis has a better prognosis. Typification by means of restriction fragment length polymorphism is very useful in clarifying the origin of the cases. PMID- 12886311 TI - [Molecular genetics of astrocytomas]. AB - Astrocytomas are the most frequent group of intracranial tumors. Among them, glioblastoma is the most aggressive one. In this review we will describe the most common genetic abnormalities found out in astrocytomas. We will refer to the epidermal growth factor receptor (EGFR), p53, p16, PTEN and DMBT1 genes. We will also present certain genetic aspects that influence the progression to glioblastoma. PMID- 12886312 TI - [Eating disorders]. AB - At the present time, research into Eating Disorders is being carried out in very different areas: clinical and diagnostic, epidemiological, comorbidity, aetiopathogeny, treatment and forms of care. This paper reviews this type of pathology, and aims to update the most important data in the different fields. The epidemiological studies of recent decades would indicate a considerable increase in the rates of incidence and prevalence of both Nervous Anorexia and Nervous Bulimia. On the other hand, there is increasing confirmation of an early appearance of these disorders. Similarly, the discovery of certain physiopathological appetite markers and/or regulators opens a new path for understanding this phenomenon. Research into these problems has reached a certain consensus in aspects such as: a multidimensional consideration of the ethiopathogeny, increasingly purified diagnostic criteria, care and therapeutic procedures and resources to be used in treatment. PMID- 12886313 TI - [Surveillance report on Diseases of Compulsory Notification in Navarra. 1999]. AB - The Epidemiological Surveillance System of Navarra includes the notification of 33 transmissible infectious diseases, to which epidemic outbreaks of any aetiology and cause are added. Reporting to the system is carried out on a weekly basis by every doctor who suspects or diagnoses any of these processes. In our autonomous community, Diseases of Compulsory Notification (DCN) are reported to the Section for the Control of Infectious Diseases and Outbreaks of the Public Health Institute on a weekly basis by the doctors of Primary and Specialised Care. Subsequently, the information is sent to the National Epidemiology Centre where data from the autonomous communities is centralised. In 1999, under the heading of diseases of respiratory transmission, 59,159 cases of Flu were reported; 65% of total annual cases were reported in the first 9 weeks of the year, with a maximum in week 4 when 6,826 cases were reported. 18 cases of Meningococcal Disease were reported to the system. Sixteen cases were confirmed microbiologically and appeared in a sporadic way. With respect to the causative serogroup, serogroup C was isolated on 8 occasions. On 7 occasions Neisseria meningitidis serogroup B was isolated. By age groups, 4 cases were declared in infants of 2 years of age (Rate: 46.1 per 100,000), nine cases in children between 2 and 9 years of age (1.53 per 100,000) and the remaining 2 cases in persons aged 20 years or over (0.49 per 100,000). 15 cases of Legionellosis were declared in 1999, all under the clinical form of pneumonia. These were isolated cases in persons of middle or advanced age. No indication was found of prior tourist trips to areas of high prevalence. In the majority of cases the origin was considered to be in the community, while one outbreak was identified as nosocomial. Similarly, there was a notable increase in the declaration of cases of Paludism, with 16 cases (EI: 5.33) affecting travellers and immigrants. PMID- 12886314 TI - [Antenatal education and recognition of the onset of labour]. AB - BASIS: In recent years, there has been a progressive increase in the number of emergencies with a preterm labour diagnosis at the Virgen del Camino Hospital. In 1996 the anaesthesia service was introduced in the labour ward. Taking into account that no other factors have changed in this service, it is believed that its introduction might have contributed to the increase of preterm labour emergencies, as well as to a decrease in the number of pregnant women who attend antenatal classes. The aims of the present study are: to check whether there are any differences with respect to attendance of the antenatal classes and the number of pregnant women who come to the hospital in preterm labour or not, and to determine the reasons why pregnant women do not attend these courses. METHODS: A case control retrospective study. The cases were women at the term of their pregnancy who were attended at the Virgen del Camino Hospital in 1998. The sample was obtained by simple random sampling, using the registries of emergencies and the labour ward. The size of the sample was 150 cases and 150 controls, who were interviewed by telephone in order to complete the data collection questionnaire. Chi square was applied for the comparison of proportions and Student's t test for the comparison of means. CONCLUSIONS: It was found that pregnant women with preterm labour were less likely to attend to antenatal classes (p=0.02; OR=0.56;CI 0.33-0.96). Attendance of antenatal classes had a positive effect for the recognition of a suitable time for coming to the hospital because of the onset of delivery, with a reduction in preterm labour emergencies, and a reduction in the corresponding workload. PMID- 12886315 TI - [Immigrant carers and elderly persons]. PMID- 12886316 TI - [Genetic alterations induced by anti-tumour treatments in paediatric patients with cancer: chemical carcinogenesis]. PMID- 12886317 TI - [Study of the accumulation of lead in the primary teeth]. AB - The purpose of the study was to determine the accumulation of lead in deciduous teeth in children living in the area of Pamplona (Spain). In this manner, we tried to make a relationship between the quantity of lead accumulated in the tooth against certain factors of exposure that were documented on a questionnaire carried out at the time that the tooth was presented. We analysed 457 deciduous teeth using a technique of microwave digestion follow by Atomic Absorption Spectrometry (AAS), to determine the concentration of lead in the sample. The mean lead concentration was 2.60+/-1.36 microg/g (range 0.25-10.71 microg/g). The lead concentration in our study is inferior to those observed in other European studies. PMID- 12886318 TI - [Analysis of the involvement of the tumour suppressor genes TP53, p16INK4, p21WAF1, RB1 and the drugs metabolizing enzymes in the development of bone tumours in children]. AB - BACKGROUND: Several tumor suppressor genes such as p16INK4, TP53, RB1 y p21WAF1 are involved in cell cycle regulation in response to DNA damage and belong to the complex pathway that regulates cell proliferation and/or differentiation. We have investigated the presence of mutations in those genes and polymorphisms of Drug Metabolizing Enzymes that could be involved in the development of pediatric bone tumors or in their outcome. MATERIALS AND METHODS: By means of PCR-based techniques, we have analyzed the presence of variations in the coding sequence of p16INK4, TP53, RB1 y p21WAF1 and of the Drug Metabolizing Enzymes in a group of 82 osteosarcomas and 47 Ewing's sarcomas as well as in a control group of 115 healthy children. RESULTS: We detected mutations of the TP53 gene in about 25% of the samples analyzed, most frequently in association with tumors of poor prognosis or reduced survival. The p16INK4 gene was homozygously deleted in 18% of the osteosarcomas, also associated with poor prognosis and unfavourable histologic subtypes; RB1 was altered in 21% of the osteosarcomas. We did not detect relevant associations between polymorphisms of the Drug Metabolizing Enzymes or mutation of the p21WAF1 and development of pediatric bone tumors. CONCLUSIONS: Alteration of TP53, p16INK4 and p21WAF1 seems to be involved in the development of pediatric bone tumors and to be an unfavourable prognostic factor in this type of tumors. PMID- 12886319 TI - [Pulse rate of the venous wave in the eco-Doppler study of the lower extremities as a sign of elevated pressure in the right atrium]. AB - AIM: The aim of this study is to determine the nature of venous flow in the lower extremities, as well as to correlate the velocity of the flow with average vein pressure in the right atrium. MATERIAL AND METHODS: Over a period of one year 236 pulsated Doppler echographs were made of patients, bearers of a venous catheter located in the right atrium. The patients were in a supine lying position and breathing gently. The lines of the Doppler wave were used to analyse the frequency of the wave, the components of its velocity, the relationships of velocity and the existence of pulsatile flow. These parameters were compared with the pressure in the right atrium. RESULTS: The study showed a cardiac and respiratory periodicity of the venous wave. A statistically significant correlation (p<0.0001) was found between average venous pressure in the right atrium and the following variables: the systolic atrium wave (a), the diastolic atrium wave (d), the relation of pulsatility (RP=Velocity min/Velocity max) and the index of pulsatility (IP=[Velocity max-Velocity min]/Velocity average). CONCLUSION: There is an inverse and significant relationship between pulsatile flow and atrium pressure. Nonetheless, although a relation exists between the different components of the venous wave, an elevation in central vein pressure cannot be predicted. PMID- 12886320 TI - [Visual synthesis of speech]. AB - The eyes can come to be the sole tool of communication for highly disabled patients. With the appropriate technology it is possible to successfully interpret eye movements, increasing the possibilities of patient communication with the use of speech synthesisers. A system of these characteristics will have to include a speech synthesiser, an interface for the user to construct the text and a method of gaze interpretation. In this way a situation will be achieved in which the user will manage the system solely with his eyes. This review sets out the state of the art of the three modules that make up a system of this type, and finally it introduces the speech synthesis system (Sintesis Visual del Habla [SiVHa]), which is being developed in the Public University of Navarra. PMID- 12886321 TI - [Update on melanoma: incidence, development and biological aspects]. AB - Melanoma is a malign tumour, highly metastatic, that has undergone a spectacular increase in all of its clinical forms in the last 40 years, with a threefold increase in its incidence. Besides genetic predisposition, intense exposure to sunlight by light skinned patients, together with antecedents of sunburn during childhood, constitute the principal risk factor for its appearance. Many clinical and histological observations suggest a clear interaction between the immune system of the host and skin cancer. In the disease's progression, in the passage from a phase of horizontal growth to an invasive phase of vertical growth--the determinant factor in prognosis--factors intervene that are only partially understood. Angiogenesis, or the development of new vessels, is necessary to maintain the growth, invasion and metastatic capacity of a tumour. VEGF and bFGF are outstanding amongst the pro-angiogenic cytokines secreted by the skin cancer cells; these favour endothelial proliferation and the invasive capacity of the tumour. Similarly, it has been found that the presence of Th1 cytokines in the "tumourous environment" prevents the disease's progression and favours regression, while a Th2 pattern favours invasion and increases the potential for metastasis. The decline in the immune response of the host might be mediated by the synthesis of immuno-suppressor cytokines by the tumourous cells (IL-10, TGF beta, VEGF) and by the expression on its surface of Fas-L molecules, which make skin cancer a privileged tissue from an immunological point of view. PMID- 12886322 TI - [Dr. D. Nicasio Landa, official epidemic doctor during the cholera epidemic of 1854-1855]. AB - In this unpublished work from 1861, which we have transcribed, Dr. Nicasio Landa (1830-1891) is revealed to have been at the forefront in carrying out epidemiological studies in Spain. He traced the medical topography, or geographical pathology, of cholera by provinces, by means of calculating the incidence rate ("millesimal proportion of those infected") and lethality rates ("millesimal proportion of the dead") using the official data on the sick and deceased in the cholera epidemic of 1854-1855, published by the General Directorate of Health and Charity, as well as the data of the population census of 1857. He worked with an infectionist conception and, besides tracing the epidemiological map, he proposed an association of cholera with the geographical constitution of the terrain in order to explain the anomalous distribution of the disease. That same "anomalous" distribution was repeated in the cholera epidemic in Spain in 1885, and in that of 1971, which is in accordance with the environmental characteristics of certain terrains that make possible the maintenance and persistence of the vibrio cholerae, according to recent research explaining the varied endemo-epidemic distribution of cholera. PMID- 12886323 TI - [Readability of informed consent]. AB - A comparative study has been made of the readability of informed consent form of the Navarra Health Service, using the Flesh and SMOG formulas. Signed editorial articles from the local newspapers Diario de Navarra and Diario de Noticias were used as a term of comparison. The results show that the readability of informed consent form is -4 (with a second paragraph of -9), while the editorial articles showed a comprehensibility of 22. The text of the second paragraph of the informed consent form is barely comprehensible, and alternative texts that are more easily comprehensible must be studied. PMID- 12886325 TI - [Adolescent mothers]. AB - Pregnancy during adolescence can be considered a health problem because it involves a greater risk, according to the WHO, for the health of mother and child. In this retrospective study an analysis was made of the socio-economic profile and the evolution of gestation and delivery of adolescent mothers attended by the Virgen del Camino hospital. A differentiation was made between two groups, with ethnic origin (Caucasian/Gypsy) taken as a variable, since it was considered that this determines the outcome of pregnancy. It was concluded that a high percentage had a partner and were under obstetric supervision and, probably for this reason, had had spontaneous deliveries, with a low index of prematurity. An association was also drawn between precocious maternity and urban environment or academic-occupational inactivity. PMID- 12886324 TI - [Economic evaluation of routine vaccination of 15 month old children against chicken-pox-zoster]. AB - BACKGROUND: An economic, cost-effectiveness evaluation was carried out that compared a hypothetical program of routine mass vaccination against the chicken pox-zoster virus in children aged 15 months in the Foral Community of Navarra against the present strategy of vaccination that is restricted to the high risk population. MATERIAL AND METHODS: Decision trees based on Markov models were used to calculate the costs of the health care of cases of infection and the costs of the effects of the vaccination program. The efficacy of the vaccination is 90 95%, and the scenario produces an immunogenicity of at least ten years, with a coverage of 90%. Account was taken of both the direct costs of health care and the indirect costs, with 1995 Pesetas taken as a constant, due to the loss in productivity of a family member, and a social view point was adopted for evaluating the study RESULTS: The index of cost-effectiveness reflects the additional cost or saving for each case of avoided infection brought about by vaccinating the children in comparison with vaccinating only those persons belonging to the high risk population sectors. The cost per avoided case is situated between 3,500 Ptas and 4,000 Ptas. For each Peseta invested in the vaccination program there would be a reimbursement of 0.45 Pesetas. CONCLUSIONS: The routine vaccination program produces an incremental cost. Only in the case of a reduction in the price of the vaccine by more than 50% would the cost effectiveness index offer a net social profit. PMID- 12886326 TI - [Erythropheresis of the single donor: study of 50 procedures and evaluation of their efficacy in reducing leucocytes]. PMID- 12886327 TI - [Commentary on the article: The use of antimicrobial drugs in the Internal Medicine Service of a general hospital: prevalence study]. PMID- 12886329 TI - [Peritoneal pseudomyxoma]. PMID- 12886330 TI - [The occupational health plan of Navarra]. PMID- 12886331 TI - [Adrenomedullin: a new peptide with many clinical implications]. PMID- 12886332 TI - [Antimicrobial drug use in the department of internal medicine of a general hospital: prevalence study]. AB - OBJECTIVE: To ascertain the general pattern of hospital antimicrobial use, costs, and adequacy of treatment in the department of internal medicine of a general hospital. MATERIAL AND METHODS: A prevalence study was carried out and all department beds were visited. Every patient who was being treated with antimicrobials during or 24 hours prior to the visit was selected for the study. Treatment was considered adequate when indication, selected drug, dosage, and treatment duration were all adequate. When considering costs, only the price of the antimicrobials was evaluated. RESULTS: We identify 173 hospitalized patients, 79 (45.6%) of them were treated with 96 antimicrobial agents. The prevalence of community-acquired and hospital-acquired infections was 39.3% and 8.1%, respectively. Empirical use amounted to 77%. Overall, b-lactams antibiotics were the most frequently used (63.5%). In 22 (27.8%) patients treatment was judged inadequate, the most frequent error being an excessive duration (17.8%). The daily spending on antimicrobials was 187,750 ptas., representing 1,085 ptas. per hospitalized patient and day. CONCLUSIONS: A high percentage of hospitalized patients receive treatment with antimicrobial drugs. Because of the low rate of antimicrobials with a specific indication, the long antibiotic course duration, the incorrect drug choice, and even more the antimicrobial prescription in non infected patients, strategies to improve the quality of antimicrobial use are clearly required PMID- 12886333 TI - [Functional activity of auditory cortex studied with SPECT: methodology validation and application in bilateral profound deafness]. AB - To evaluate the differences in the functional activity of the auditory cortex between normal hearing and profound deafness, a perfusion single photon emission tomography (SPECT) study was designed. SPECT stereotaxic localisation of the auditory cortex was previously validated in 2 brains by means of an anatomical study of the macroscopic localisation and cytoarchitecture of the auditory cortex. Additionally, 15 controls with normal hearing and 30 patients with profound bilateral deafness were scanned using external anatomical point sources (glabela, ineon) for stereotaxic location of the auditory cortex. The normal controls were scanned in auditive deprivation and, in 10 cases, during a monoaural tonal stimulation. Cerebral blood flow relative to cerebellum (relCBF) was assessed in the auditory cortex. The anatomical study showed that mean differences between the true auditory cortex size and the measured SPECT value were less than 2.5 mm. Nevertheless, only the caudal aspect of this area corresponded to the primary auditory cortex in the cytoarchitectonic study. During tonal stimulation, control subjects presented a significant increase of relCBF in the auditory cortex bilaterally, with significant differences in the asymmetry index (contralateral to the side of stimulation). The relCBF in the auditory cortex of controls in deprivation conditions was significantly higher than in deaf patients. There were no significant differences between groups of deaf patients, however the highest values were seen after cochlear implant. SPECT is a suitable method for studying changes in auditory cortex activity relative to different functional conditions, with a possible role in cochlear implant candidates in predicting the future benefit of the implantation. PMID- 12886334 TI - [The main problems of occupational health in Navarra, 1994-1996]. AB - In 1996 the Navarra Occupational Health Institute, an autonomous technical organization that integrates the functions and means related to occupational health matters, drew up the Occupational Health Diagnosis of Navarra, which is a region characterised by its important industrial sector and its high proportion of small and medium size companies. The results emphasise the high accident rate in the construction sector, the constant increase of professional diseases through "fatigue of tendon pods", as well as the importance of osteo-muscular processes as a cause of temporary and permanent disability. Forty sentinel occupational tumors, mainly mesothelioma, and 27 Obligatory Statement Diseases of occupational origin, mainly brucellosis, were identified in a period of seven years and three years, respectively. Verification was made of a lack of systematised information about population exposure to occupational risks, a scarce development of medical surveillance of exposed workers, and a lack of information about the incidence of occupational injuries in self-employed workers. PMID- 12886335 TI - [Treatment of fibromyalgic syndrome with feedback]. AB - Fibromyalgic syndrome is one of the most frequent chronic pain disorders in everyday clinical practice. At present, its treatment is restricted by lack of knowledge about its aetiology. This paper describes training in Biofeedback Electromyography as a possible treatment based on the alterations of muscular activities shown by these patients. A review is made of the empirical evidence of its use so far in relation to this syndrome. PMID- 12886336 TI - [Leptin and body weight regulation]. AB - The discovery of leptin, besides causing a revolution in physiological knowledge about the regulation of body weight, awakes an interest in fat tissue as an endocrine organ and its active contribution to the establishment of energy deposits. Leptin, principally secreted by the adipocytes, interacts with hypothalamic receptors, and is capable of activating diverse neuroendocrinal routes responsible for controlling the balance between energy intake and expenditure. Like other cytokines, leptin is also able to play a role in the complex regulation of diverse metabolic aspects. Since the discovery that the placenta, gastric mucous or the star-shaped cells of the liver (Kupffer cells), are places that produce leptin, a great range of functions have been attributed to this hormone; since it appears to favour foetal development and can act as a factor of satiety in the stomach or participate in the afferent signals mediated by the vagus. Besides, leptin participates es in the immune and inflammatory responses, in reproduction, in processes related to angiogenesis, control of arterial pressure, lipolysis, etc. This paper reviews the role of leptin in the regulation of body weight, as well as different aspects related to obesity. PMID- 12886337 TI - [Considerations on socio-health care]. PMID- 12886338 TI - [Incidence of cancer in Navarra]. AB - The data on the incidence of cancer in Navarra for the two year period 1993-94 is set out by age groups and sex, with the aim of making available the most recent information contained in the Navarra Cancer Register. The collection of data for the years 1993-94 was carried out in an active manner in the information sources of the register. The procedures of previous years regarding collection, control of duplicates and processing were maintained. Both the quality indicators and the calculations of the different rates were obtained following the recommendations proposed by the International Agency for Research on Cancer (IARC). In the 1993 1994 period, 5.186 incident cases of malign tumours were registered in Navarra. Some 59% occurred in men. Excluding nonmelanoma skin cancer, the crude rate of incidence was 475 in men and 319 in women; the rate adjusted to the world population was 297 per 100,000 for men and 186 per 100,000 for women. If nonmelanoma skin cancer is excluded, 55% of all of the cancer cases diagnosed during 1993-1994 in men were located in the lung, prostate, bladder, colon and stomach. In women they were located in the breast, body of the uterus, colon, lymphoid tissue and stomach, representing 56% of the total cases. PMID- 12886339 TI - [AIDS/HIV epidemiological surveillance in Navarra]. AB - The incidence of AIDS in the European Union has been declining since 1996 as a result, at least in part, of the use of more efficient treatments for HIV infection. The same tendency can be observed in Navarra. In 1998 the incidence of AIDS in Navarra, 75 cases per million inhabitants, was situated at an intermediate level in the ranking of Spanish autonomous communities. In the same year, the highest rates of incidence in Europe were recorded in Spain (93.3 cases per million inhabitants) and Portugal (81.8), and the lowest in Ireland and Finland, with less than 5 cases per million inhabitants. With respect to the incidence of HIV infection, 2,240 cases were diagnosed in Navarra between 1985 and December 1998. The available data indicate that at least 1,864 of these cases were resident in Navarra. The annual number of new diagnoses of HIV infection show two high points in the years 1987 and 1991, with over 200 annual cases. From 1993 onwards, a progressive decline can be observed in the annual number, with 171, 145, 19, 86 and 63 cases in the final year, 1998. The figure of over 2,000 cases of HIV infection contrast with the 694 cases of AIDS registered in this autonomous community in the same period and help to resituate the epidemic in more accurate terms. The development of AIDS in 40% of the persons with HIV infection and the death of 26% have been confirmed, although lethality has been declining in recent years. The use of intravenous drugs in 73.2% of the AIDS cases is the most frequent category of transmission. 13.4% are due to transmission by heterosexual contact and 6.1% to homosexual practices between males. Although the cases of AIDS have been declining in Navarra since 1996, the data from the microbiology laboratories and from the hospital services that treat persons with HIV infection indicate that a considerable number of new infections will continue to occur, which justifies the need for maintaining the prevention programs in order to avoid new cases of a disease whose prognosis is of such seriousness PMID- 12886340 TI - [Evolution of the prevalence of bedsores in the Hospital of Navarra]. AB - INTRODUCTION: The rates of incidence and prevalence of patients with sores in hospital are indicators of nursing care quality. The aim of this study is to evaluate such prevalence in the last two years (1997-1998), comparing the patients with sores, the evaluation of the risk of developing sores, and the factors associated with their development. MATERIAL AND METHODS: A transversal study was made, in which the subjects under study were all of the patients hospitalised in the Hospital of Navarra on the day set for the study. The study gathers the variables of sex, age, unit of hospitalisation, date of admission and date of the study, presence and number of sores, their development in or outside the hospital, evaluation of risk on admission and evaluation of risk on the day of the study (according to the modified Braden scale). RESULTS: The prevalence of patients with sores was 8.9 per 100 patients hospitalised in 1997 and 5.1 in 1998. The difference is statistically significant with a value of p=0.025. Age, the classification of patients within those of risk, and the time of hospital stay are factors related to the development of sores. CONCLUSIONS: The factors associated with the development of sores detected in the study lead us to be more attentive in our vigilance of the patients that show them. The follow up of the prevalence and incidence of bedsores in hospitals is an instrument that makes it possible to continue improving understanding, prevention and treatment of such lesions in patients. PMID- 12886342 TI - [The challenge and the skate of the public health services]. PMID- 12886341 TI - [Heart attack in the context of a hyperthyroidism. On one case. ]. PMID- 12886343 TI - [Health in a century of Spanish history (1898-1998). One hundred years later]. PMID- 12886344 TI - [Positron emission tomography with F-18-FDG: a new tool in the evaluation of patients with head and neck tumors]. AB - INTRODUCTION: Positron emission tomography using fluoro-deoxyglucose (PET-FDG) imaging has been shown to be effective in detecting and staging malignancies based on tumor glucose metabolism. The aim of the study was to evaluate the use of PET-FDG for the detection of metastatic lesions as well as early recurrence in patients with head and neck tumors. MATERIAL AND METHODS: Eleven patients were examined with PET-FDG to study the reliability of PET in assessing regional nodal status and in identifying distant metastasis (group I) and 37 patients who had previously received curative treatment and who presented differential diagnostic problems were imaged to differentiate between scar and residual or recurrent cancer (group II). PET-FDG studies were compared to results of computed tomography (CT) in 35 patients, magnetic resonance imaging (MRI) in 4 patients and both techniques in 6 patients. RESULTS: All PET-FDG studies were positive in group I, while CT failed to detect metastatic lesions in three patients. In group II PET-FDG accurately detected recurrent disease in 22/25 patients, while CT/MRI were negative in 4 cases and equivocal in 6 cases. However, there was a false positive PET study with equivocal CT in a patient with local infection. CONCLUSION: PET-FDG was highly effective in detecting metastatic cervical lymph nodes in head and neck tumours. It was most helpful in differentiating residual or recurrent tumour from scar sequelae and it enhanced the diagnostic accuracy when CT and MRI were equivocal due to anatomical distortions. PMID- 12886345 TI - [Self-image with regard to weight in a group of adolescent girls]. AB - The explanatory models generally accepted at the present time concerning eating disorders, particularly anorexia nervosa, are the multidimensional ones. Different factors have been put forward such as predisposing factors, facilitators, triggering and sustaining factors of Anorexia nervosa. Facilitators and triggering factors include those with a sociocultural origin such as social pressure to have a certain look, closely related to a low body weight. This study aims to discover the self-image of a group of female teenagers with respect to their weight, as well as to ascertain which attitudes derive from self-image. To this end a survey was conducted in two high schools in Tudela in June 1998. A hundred and thirty teenagers from 15 to 17 years old attending compulsory secondary education were surveyed. The results indicate that girls from this group have a negative self-image with respect to their body weight; most of them think they should be thinner and that is the reason why they change their behaviour in order to lose weight. PMID- 12886346 TI - [Evaluation of the Program for the Prevention and Control of Tuberculosis during the period January 1993 - June 1996]. AB - In 1992 a Technical Commission was formed in Navarra with the participation of different specialists who drew up the Program of Vigilance and Control of Tuberculosis. We present the results of the evaluation of this program for the period from January 1993 to June 1996. In the 3.5 years of the study, 419 cases of tuberculosis were found, 317 belonging to respiratory forms and 102 to extra respiratory forms. The annual rate of incidence of total tuberculosis, 22.8 cases per 100,000 inhabitants observed in Navarra, is the lowest of those registered at the level of the Autonomous Communities according to the TIR study of the SEPAR (acronym for Spanish Society of Pneumology and Thoracic Surgery) in 1996, and the second lowest, after Castilla-La Mancha, according to the multicentric study of the National Centre of Epidemiology. In Navarra the highest rates of incidence are produced in those over 65 years of age, following the pattern observed in the more developed countries. 13.9% of the patients with respiratory tuberculosis showed co-infection by HIV, and besides more than 90% of these were users of intravenous drugs. The percentage of immigrants, prisoners and the destitute observed in the Navarra series is lower than that found in areas such as Madrid, Barcelona or Zaragoza. 91% of the cases of respiratory tuberculosis showed bacteriological confirmation and 98% of the cases were diagnosed and treated in specialised care. Outstanding were the high follow-up of the cases until discharge and the high percentage of cures achieved, some 85.4%. PMID- 12886347 TI - [Pedunculopontine tegmental nucleus. Anatomy, functional considerations and physiopathological implications]. AB - Pedunculopontine tegmental nucleus is formed by an ensemble of cholinergic and non-cholinergic neurons located in the caudal pontomesencephalic tegmentum, surrounding the superior cerebellar peduncle. It is an integral part of the reticulate formation of the brain stem, with extensive anatomical connections and highly varied functions. By means of ascendant projections that it sends to the thalamus, it intervenes in the waking-sleep cycle. Besides, it constitutes the most caudal nucleus of the neuroaxis, receiving connections from the basal ganglia, for which reason it has attracted the interest of those researchers concerned with the study of these structures. Thanks to its reciprocal connections with the basal ganglia, as well as to its descending projections to different structures of the pons, medulla and spinal cord; it has been related to the control of locomotion. Recently, it has also been considered as a possible centre for the integration of the motor information provided by the dorsal striatum with the motivational or limbic information proceeding from the ventral striatum, to permit its direct access to bulbar or spinal motor centres. In this work we will review its anatomical and functional characteristics, as well as its implication in some diseases of the nervous system such as narcolepsy, progressive supranuclear paralysis, schizophrenia and Parkinson's disease. PMID- 12886348 TI - [Non-steroid anti-inflammatory drugs and adverse gastrointestinal effects. An unresolved problem]. AB - Non-steroid anti-inflammatory drugs are a class of medicine widely used throughout the world. This is a pharmacological group in continuous growth, to which some new molecules have been added in recent years. The great drawback of non-steroid anti-inflammatory drugs are their adverse effects, outstanding of which due to their frequency and importance being those that occur in the gastrointestinal tract. By means of a search in Medline and other databases, this work reviews the latest data published on the incidence of dyspepsia, gastroduodenal lesions, gastrointestinal complications and mortality associated with consumption of non-steroid anti-inflammatory drugs. Similarly, a brief description is made of the mechanism of lesions to the stomach of the non-steroid anti-inflammatory drugs and the different risk factors that condition the appearance of adverse effects at the gastrointestinal level. Finally, an analysis is made of the preventive strategy and the different medicines that can be used to this end and a contrast is made of the evidence extracted from the different published studies and the reality of the use of the different "gastroprotectors". This review concludes with a series of questions that still remain unresolved concerning treatment with non-steroid anti-inflammatory drugs and their lesions to the stomach. PMID- 12886349 TI - [Light and shadows in the medico-social fight against tuberculosis. A retrospective look at tuberculosis in Pamplona (XX Century)]. AB - Tuberculosis is one of the infectious diseases that can be qualified as constant throughout the history of illnesses. Nonetheless, starting in the early decades of the XX century, mortality provoked by this complaint began a progressive decline throughout Europe. This retrocession also took place in Spain, although with evident regional and local differences. Because of its nature, throughout this century a series of social and medical mechanisms and strategies have been developed around it. Together with the undoubted importance of quantitative analysis, it is essential to pay attention to the social, economic and medical care factors that have affected its evolution, and the degree of effectiveness of the anti-tuberculosis campaigns. In this way we will be in a situation to present the results - those lights and shadows - of the socio-care policies created for the country as a whole, but applied to a specific space and society such as Pamplona. Similarly, it is not possible to disregard the renewed interest that is being shown in different forums because of the resistances and variations that have been detected in the bacilli, and especially because it has been related to the end of the century "plague", AIDS. PMID- 12886350 TI - [Prevalence of smoking, awareness and evaluation of cessation methods amongst medical students]. AB - This work describes and analyses the prevalence of smoking amongst medical students, their awareness of the methods of smoking cessation and the effectiveness they attribute to these methods and to medical advice. A questionnaire was distributed to students of medicine at the University of Navarra with open questions on the methods of smoking cessation of which they were aware, the effectiveness attributed to each of these and the effectiveness attributed to medical advice. Information was also gathered on: sex, year of study, smoking habit, average number of cigarettes smoked per day and the length of exposure to tobacco. Of 480 valid interviews, the prevalence of active smokers was 27.5% (CI 95%: 23.5-31.9); 4.2% ex-smokers and 68.3% who had never smoked. 42.7% named nicotine patches as a method of cessation and 25.2% nicotine chewing gum. With respect to the effectiveness attributed to each method, a median and interquartile range (IQR) of 20% (0-50) for the nicotine patches, 10% (0-40) for nicotine chewing gum and 20% (1-50) for medical advice. The more senior students and those that had been smoking for longer mentioned nicotine patches more frequently; more women than men knew about nicotine chewing gum. The senior students attributed greater effectiveness to nicotine patches. A future reduction is predicted in the number of active smokers in the medical sector. Amongst medical students the effectiveness of medical advice is overestimated. It is necessary to increase awareness of the methods for smoking cessation amongst medical students. PMID- 12886351 TI - [Situation of Diseases of Compulsory Declaration (DCDs) in Navarra. 1998]. AB - The System of Epidemiological Vigilance of Navarra comprises the notification of 33 transmissible infectious diseases, to which are added epidemic outbreaks of any etiology or cause. Notification to the system is carried out on a weekly basis by any doctor who suspects or diagnoses any of the processes. In our Autonomous Community, the Diseases of Compulsory Declaration (DCDs) are reported to the Section of Vigilance and Epidemiological Control of the Institute of Public Health, on a weekly basis by the doctors of Primary and Specialised Care. Subsequently, the information is sent to the National Centre of Epidemiology, where data from the Autonomous Communities is centralised. In 1998, with respect to the heading of diseases of respiratory transmission, the only disease that showed an epidemic index higher than 1 was Flu, with 44,666 reported cases, (Estimated Incidence: 1.15), and which has a clear seasonal component nin the first 8 weeks of the year 67% of the cases corresponding to the yearly total were reportedn with a maximum in week 4 with 6,361 reported cases. Fifteen cases of meningococcal disease were diagnosed, (Rate 2.88 cases per 100,000 inhabitants), lower to that declared in 1997 and the second lowest in the last 25 years after the rate of 2.31 in 1994. All of the cases were confirmed microbiologically and appeared in a sporadic way. With respect to the causative serogroup, on 9 occasions serogroup C was isolated, with the latter responsible for the death of a girl and an elderly woman. On 6 occasions Neisseria meningitidis serogroup B was isolated. By age group, 4 cases were declared in children under 2 years of age (46.1 per 100,000), nine cases in children between 2 and 19 years of age (8.65 per 100,000) and the two remaining cases in persons of 20 years or above (0.49 per 100,000). The predominant clinical form was sepsis in 9 cases and meningitis in the 6 remaining cases. Six cases of Legionnaire's disease were declared in 1998, all under the clinical form of pneumonia; these were isolated cases of middle-aged persons resident, in five cases, in different localities in the south of Navarra without any antecedents of travelling. PMID- 12886352 TI - [The Navarra Health Service-Osasunbidea declares the "San Francisco Javier" Psychiatric Hospital to be a Psychogeriatric Centre]. PMID- 12886353 TI - [Pulmonary bronchogenic tuberculosis with respiratory distress of the adult and haemophagocytic syndrome with fatal evolution]. PMID- 12886354 TI - [Clinical and demographic aspects of Parkinson's disease]. PMID- 12886355 TI - [Public Health and the health system]. PMID- 12886356 TI - [Family medicine as an area of knowledge in the Spanish university]. PMID- 12886357 TI - [Inverse association between smoking and idiopathic Parkinson's disease]. AB - An evaluation is made of the effect of smoking during the period of life between 20 and 50 years of age and idiopathic Parkinson's disease in a case-control study carried out in Navarra. An inverse association is observed: for the number of smokers (OR = 0.66, CI 90% 0.41-1.05); for the number of years smoking (OR =0.22, CI 90% 0.05-0.97); and for the quantity of cigarettes smoked (z = 2.2298, p = 0.02). Likewise, a dose-response relationship exists between the number of cigarettes consumed and the risk of Parkinson's disease (p = 0.01). The same effect is maintained on analysing the data by matching 1:1 (OR = 0.59, CI 90% 0.31-1.10). All of this contributes to giving consistency to the protective effect of smoking against idiopathic Parkinson's disease. The anti-oxidant status, measured by means of different parameters in peripheral blood, is lower in the cases than in the tests for reduced glutathione (GSH) (p = 0.001) and is an independent variable with respect to smoking. PMID- 12886358 TI - [Prevalence of cardiovascular risk factors in the working population of Navarra]. AB - A high prevalence of cardiovascular risk factors (RFs) has been reported in Spain. The aim of the study was to determine the prevalence of the principal RFs of coronary heart disease (hypercholesterolaemia, low HDL, smoking, arterial hypertension, hyperglycaemia, obesity and sedentary behaviour) in the working population of Navarra. The present study represents the first baseline evaluation of a randomised intervention of primary prevention. The sample under study consisted of 790 workers (742 men and 48 women). Information was gathered on each worker by means of a standardised questionnaire, by the company medical service, and a physical check up was carried out. Total cholesterol and HDL were measured in venous blood by enzymatic autoanalyser. A description was made of the data (percentages of the risk factors and estimation of means for continuous variables). 67.2% of the sample had total cholesterol values higher than 200 mg/dl and 26.3% had HDL levels below 35 mg/dl. 37.1% were regular smokers. The prevalence of high blood pressure was 7.6%. 72.1% reached a body mass index above 25 Kg/m2 and 20.6% classified themselves as sedentary as they did not participate in any physical activity during their leisure time. The prevalence of hyperglycaemia was 1.4%. Only 4.2% of the workers were free of the risk factors under study. A high prevalence of hypercholesterolaemia was detected, and overweight, which supports the pertinence of establishing intervention programs of primary prevention of cardiovascular diseases at the work-site. PMID- 12886359 TI - [Epidemiological profile of patients infected by Human Immunodeficiency Virus (HIV)]. AB - INTRODUCTION: The aim of the present work is to identify the principal characteristics of a sample of individuals at the moment of diagnosis of HIV infection. MATERIAL AND METHODS: Descriptive, retrospective study, based on the hospital clinical records of 70 HIV+ patients, without AIDS, selected by means of simple aleatory sampling. RESULTS: Transmission categories: Users of Injectable Drugs (UID) 81.4%, heterosexuals 10%, homo/bisexuals 4.3% and transfusions/plasma donors 2.9%, sex: ratio man/woman = 3.8/1, average age on diagnosis: 27.3 +/- 7.0 years (UID 26.3 +/- 5.1 years, heterosexuals 29.6 +/- 2.1 years, homo/bisexuals 27.3 +/- 3.9 and transfusions/plasma donors 51.4 +/- 23.1 years (p = 0.02). Heterosexual transmission amongst women was 18.8% against 7.4% in men (2.5:1). Marital status: single in 66.7% of the cases. Working activity: 48.9% without work, outstanding amongst those with work was the relatively high proportion of HIV+ working in the building trade (29.2%). Smoking habit: 91.8% were smokers, with greater incidence amongst the UID (100%). Clinical situation: during the five years of follow up, 49% of the patients developed AIDS, with cachectic syndrome because of HIV being the first disease indicative of AIDS at the time of its appearance in the majority of the cases (36.7%) in this sample. The average interval between the date of diagnosis of HIV infection and diagnosis of AIDS was 44 months (rank: 0.9-131). CONCLUSIONS: HIV infection in our sample appears mainly in individuals who are young, male, UID, unemployed and smokers. Prevention strategies should be directed at women, at individuals with a higher risk of heterosexual transmission and at youths who are unemployed or low skilled. PMID- 12886360 TI - [The impact of technology in Pathological Anatomy and the contribution of this speciality to technological development]. AB - It is obvious that technology has contributed throughout history to the development of the different sciences. In this article, we define the concept of Pathology as a medical speciality, and we explain its influence in a hospital, considering very different fields such as education, research, quality control, hospital information, and patient care. This speciality has undergone a considerable evolution, to which technological innovation has undoubtedly contributed. As a basic discipline, it is of great importance in pre and post graduate training, in the medical education at the hospital or outside it, and in the fields previously mentioned. Its relation with other disciplines such us Chemistry, (fixation and dyeing), Physics (mechanical devices), Mathematics (algorithms, morphometry, statistics...) and Telecommunications (telepathology, image analysis...) is examined and their contribution to Pathology is evaluated. We are also aware of contributions made by Pathology to technological innovation in the evaluation of different diagnostic methods or in the recent therapeutic technologies based on Radiotherapy, Hyperthermia, laser, prothesis, etc.; where histological examination provides accurate information about the therapeutic capacity or side-effects, or the rejection reactions caused, aiding the research to obtain adequate results. PMID- 12886361 TI - [Continuous quality improvement in the control of the type2 diabetes in Navarra. GEDAPSNA study 1996]. AB - Diabetes mellitus type 2 constitutes one of the principal public health problems today, representing an important burden on care and involving extremely high economic costs, both direct and indirect. To provide an adequate response from the different public health systems, the St. Vincent Declaration was drawn up containing certain recommendations on the protocol of care and the carrying out of controls of continuous quality improvement, since it is a chronic pathology that requires a proactive health policy, which prevents, and provides early detection of its complications. In the present work we set out the results of the first study carried out in Navarra on the indicators of quality in the care provided to diabetics in Primary Health care, carried out by the Study Group on Diabetes Mellitus in Primary Care (GEDAPSNA). The study was made of an aleatory sample of 496 patients, by means of gathering data about their clinical records for the year 1996. 12 process indicators and 12 indicators of intermediate and final results were used. The results detect some important deficiencies that require early intervention: lack of exploration and management of the lesions that make up so-called diabetic foot; low achievement of results in the control of arterial hypertension in this group of patients; and absence of exploration of the lesions of the fundus of the eye in more than 50% of type 2 diabetics. PMID- 12886362 TI - [Legal disability in patients with mental disorders]. AB - This article offers an approach to the present juridical situation of the processes of occupational incapacity - in the contributive and non-contributive types - for patients with psychological disorders, taking into account the wide range of nuances that this implies. Finally, some verdicts of the Spanish Superior Court of Justice are added, in order to illustrate the present situation about the jurisprudence referring to permanent contributive incapacity. PMID- 12886363 TI - [The handling of inhalers by bronchitis patients]. AB - A high percentage of patients are admitted to the Internal Medicine Unit with respiratory pathology, principally chronic obstructive pulmonary disease. The basis of treatment of the patient with chronic obstruction of the airways are the bronchodilators and corticoids administered by inhalation. There is a close relation between the correct use of the inhaler and its therapeutic efficiency. As chronic obstructive pulmonary disease is chronic, the patient is obliged to utilise inhaler treatment both in the hospital and at home. With the aim of determining what they know about inhalers, and how they use them, as well as to obtain information about the follow up practised with these patients, a study was designed that was carried out for one month in the Internal Medicine Unit of the "Virgen del Camino" Hospital of Pamplona. Data was gathered using a questionnaire with nine questions that were put to a sample of 23 patients admitted to this unit and who used inhalers during their hospital stay. It was verified that 57% of the patients did not use the inhalers correctly, 61% did not know when the container had run out and 70% did not even know the name of the medicine they were using. The study showed that the patients did not correctly administer the inhalatory medicines, which annulled or diminished the effectiveness of the treatment, given that there is a close relation between the correct use of the inhaler and its therapeutic efficiency. PMID- 12886364 TI - [Continued education. The start of a new era]. PMID- 12886365 TI - [Anaesthetic in experimental surgery. A four year experience]. PMID- 12886366 TI - [Secondarily generalised, partial crises as an expression of multiple sclerosis]. PMID- 12886367 TI - [Placenta percreta as cause of massive haemorrhaging during uterine curettage]. PMID- 12886369 TI - [Impact for the control of cholesterolemia: diet versus drugs]. PMID- 12886385 TI - [Neuro-Behcet]. PMID- 12886386 TI - [Pulmonary toxoplasmosis: occurrence in an immunocompromised adult]. PMID- 12886387 TI - [From oral candidiasis to acidosis status]. PMID- 12886388 TI - [Body mass index and triceps skinfold correlation in children from Paulinia city, Sao Paulo, SP]. AB - Body mass index (BMI) has been considered a criterion to define and analyse obesity in adults and children. BACKGROUND: the purpose of this study was to evaluate the correlation between BMI and triceps skinfold (TSF). METHODS: there were studied 4,236 children (3.1-10.9y); 48.3%M:51.6%F, from four studies made in Paul nia, SP-Brazil. Height, weight and TSF (Holtain caliper) were measured. For each children BMI was calculated and transformed in SDS, according to North American data (Frisancho, 1993). Multiple linear regression analysis (stepwise) was used for the whole population and in three groups according to BMI: A) SDS < or = -1.0; B) -1.0 < SDS< 1.0) SDS(3) 1.0. Data were processed with SPSS software. RESULTS: in group A, the TSF (7.8 +/- 2.3) variability was lower when compared with the groups B (10.1 +/- 4.0) and C (17.8 +/- 6.2). In multiple linear regression with the whole population, R = 0.478 for TSF. In groups B and C, R = 0.364 and 0.368 respectively for TSF, and in group A it was only 0.032. CONCLUSIONS: these observations demonstrated a height correlation between BMI and TSF in children with obesity risk (group C). Therefore, we conclude that, in Brazil BMI can be used for children's research of obesity in population studies, instead of TSF. PMID- 12886389 TI - [Elevated cyclosporine A trough levels in HCV positive kidney transplant recipients]. AB - OBJECTIVE: Compare the CsA trough levels of HCV+ kidney transplant recipients to a control group METHODS: All anti-HCV positive patients that received a renal allograft between January 1992 and April 1996 were initially included as cases. Patients with diabetes mellitus, HBsAg+, who were taking medication that could modify CsA pharmacokinetics and those with elevated aminotransferases were excluded. For each anti-HCV positive index case the following transplanted anti HCV negative patient was included as a control. Third generation ELISA was used for determination of the anti-HCV status and CsA dosages were performed by polarized fluorometry with polyclonal antibodies. RESULTS: No differences in the demographic variables were found. The average CsA through levels in the first month were higher (551 +/- 280 ng/ml) in the 23 cases as compared to the 31 controls (418 +/- 228 ng/ml; p< 0.05). The differences became apparent at the end of the first week (528 +/- 275 versus 344 +/- 283 ng/ml; p<0.01) and persisted at discharge (582 +/-284 versus 457 +/- 229; p=0,08). CONCLUSION: We concluded that anti-HCV positive patients have higher blood levels of CsA for a particular dosage, than anti-HCV negative controls. Prospective studies with a more appropriate pharmacokinetic approach are needed to confirm the present findings. PMID- 12886390 TI - [Cancer in thyroid nodules: evaluation of ultrasound and fine-needle aspiration biopsy]. AB - BACKGROUND: Thyroid nodules are a very common clinical problem and fine-needle aspiration biopsy (FNAB) is its main diagnostic test. Recently, it has been shown that some ultrasonographic features are associated to an increased risk of cancer, but a comparison between FNAB and ultrasonography (US) or its combination is not available. The aim of this study was to evaluate FNAB and US performance, isolated or combined, for cancer diagnosis in thyroid nodules. METHODS: Eighty patients, with palpable solitary thyroid nodules or multinodular goiters, who were to undergo surgery, were selected among 289 patients seen in our clinic between March, 1995 and July, 1997. US and FNAB were performed by one of us. US was considered positive for cancer when microcalcifications, absent halo or hypoechogenicity were seen. FNAB was considered positive for cancer when it was either suspicious or compatible with malignancy. Both methods (FNAB and US) were evaluated in parallel combination (positive when FNAB or US was positive) and in serial combination (positive when both, FNAB and US, were positive). RESULTS: Sensitivity and specificity of FNAB and US for cancer diagnosis in thyroid nodules were, respectively, 87% and 62%, and 81% and 70%. Serial combination of FNA and US had sensitivity of 69% and specificity of 91%, and parallel combination resulted in sensitivity of 94% and specificity of 52%. CONCLUSION: The US had good accuracy in the evaluation of thyroid nodules, and can help us take some decisions in the clinical practice, supplementary to FNA. Nevertheless, the quality of the ultrasound examination depends on the experience of the examiner and on the characteristics of the equipment, so new studies are necessary to evaluate these factors to the current usage of ultrasonographic characteristics in the evaluation diagnostic of thyroid nodules. PMID- 12886391 TI - [Karyotype in acute myeloid leukemia: importance and type of aberrations in 30 patients at diagnosis]. AB - INTRODUCTION: Cytogenetics in AML at diagnosis is a well defined prognostic tool. OBJECTIVE: The authors analized karyotype (KT) and clinical data of newly diagnosed AML patients. METHODS: Thirty patients were studied, 16 male and 14 female. Age ranged from 19 to 84 years. Diagnostic criteria were based on WHO classification, immunophenotyping and G banding cytogenetics. They were treated according to standard protocol (daunorrubicin and cytarabine - 3+7) and those who had Acute Promyelocytic Leukemia additionally received ATRA. RESULTS: KT success rate was 84%. According to KT patients were divided into 4 groups: favourable prognosis (FP) (6) (t(8;21), t(15;17)); intermediary prognosis (IP) (7)(four normal karyotypes, + 8, t(1;2) and del 18(q)); unfavourable prognosis (UP); and 3 secondary AML; two evolving from prior Mylelodysplastic Syndrome and one presenting as an initial blast crisis of chronic myeloid leukemia. The median age of FP was 23 years while UP was 60 years (p<0.003). In the FP, 5/6 (83%) achieved complete remission (CR) while only 1/7 (20%)in the IP and 1/8 (12,5%) in the UP. There was a tendency of higher leukocyte count in the unfavourable group. CONCLUSIONS: The rate of karyotype aberrations in AML was 80% and in accordance to literature data (65-95%). There was a clear difference in CR rates between favourable and unfavourable prognosis group. PMID- 12886392 TI - [Evaluation of oxygenation, ventilation and respiratory mechanics before and after endotracheal suction in mechanically ventilated children]. AB - OBJECTIVES: The aim of this study was to know the effects of endotracheal suction in respiratory mechanics and oxigenation of patients with mechanical ventilation. METHODS: 13 children were studied in the pediatric intensive care unit of Hospital Sao Paulo, age between 47 days and 5 years old, male and female, surgical and clinic pathology, intubated by cuffed endotracheal tube, sedated and paralyzed few minutes before measurements, under previous established suction routine without preventive maneuvers, followed by a continuous monitoring of oxygenation, ventilation and respiratory mechanics under identical ventilatory sets. The parameters analyzed was Heart rate; SpO2; ph arterial; PaO2; PaCO2; SaO2; inspiratory and expiratory tidal volume; minute volume; dynamic compliance, respiratory resistance; mean airway pressure; PEEP and PEEPi., and the measurements were made immediately before suction, immediately after, ten and twenty minutes after suction RESULTS: The results showed that the technique increase the CO2 arterial pressures (PaCO2) even after 20 minutes; decrease the oxygen saturation (SpO2) immediately after the procedure with regular recuperation after 10 minutes and decrease the lung compliance (Cdin.) immediately after with lower recuperation after 10 minutes. CONCLUSION: We concluded that intratracheal suction in front of compromise of oxygenation, ventilation or respiratory mechanic, applied as minimal as possible under preventive maneuvers. We need more studies to establish the real need of intratracheal suction and a practice guideline of intervention to avoid deleterious effects of that in pediatric patients. PMID- 12886393 TI - [Overweight and obesity prevalence in Northeast and Southeast Regions of Brazil]. AB - OBJECTIVE: to evaluate the prevalence of overweight and obesity in children, adolescents, adults and elderlies from Northeast and Southeast regions of Brazil. METHODS: anthropometric evaluation of 17.184 people, studied in Life Pattern Research conducted by Brazilian Institute of Geographic and Statistics in 1996/97, by body mass index. Overweight and obesity definitions according to body mass index values proposed by Cole et al. which are related to adult definition. RESULTS: It was studied a little more females in this study (53.1%). Average age was 29 years and five months ( +/- 20 years), median was 25 years and six months. Overweight prevalence was 10.8% in children, 9.9% in adolescents and 28.3% among adults and obesity was 7.3%, 1.8% and 9.7% respectively. It was noticed a gradual increase in overweight or obesity prevalence since infancy through adult age with decrease among the elderly. CONCLUSION: Prevalence of overweight and obesity joined is higher among females. More than half of the women among 50 and 69 years old, from Northeast and Southeast regions of Brazil have overweight or obesity. There are few studies of overweight and obesity among children and adolescents and a lot of definition for obesity in children which makes more difficult the comparison of studies of prevalence among children. PMID- 12886394 TI - [Follicle stimulating hormone as predictor of mobile spermatozoa in testicular biopsy of azoospermic patients]. AB - OBJECTIVE: To define predictive factors of mobile spermatozoa recovery in azoospermic patients. METHODS: Testicular volume, serum follicle stimulating hormone (FSH), luteinizing hormone (LH), prolactin (PRL) and testosterone levels were assessed in 60 azoospermic patients. Patients underwent bilateral testicular biopsy with local anesthesia. Samples were classified according to absence of spermatozoa, presence of motile and nonmotile spermatozoa, and histological findings. Age, hormone levels, testicular volume and histology with motile spermatozoa recovery were compared. P < 0.05 was considered significant. RESULTS: Non-obstructive azoospermia was diagnosed in 45 patients. Significant differences were detected between the group with motile spermatozoa recovery and the group with absence of spermatozoa in terms of FSH levels (P=0.037 ANOVA one-way). A ROC curve was used to define FSH values below 16.05 IU/L (sensitivity: 76.2%, specificity: 67.7%) as predictive factors for motile spermatozoa recovery. Other statistical differences were not detected. CONCLUSIONS: FSH levels below 16.05 IU/L showed good accuracy to predict the presence of motile spermatozoa in the testicular biopsy of azoospermic patients. Physical examination, testosterone levels, LH and prolactin were not useful as predictive factors in the present study. PMID- 12886395 TI - [Quantitative parameters to evaluate the publication of scientific papers]. AB - BACKGROUND: The assessment of the publication of scientific works is a hard challenge when it is necessary to classify a curriculum vitae or even to verify the performance of an institution, such as a medical school, a postgraduate program or a research department. Most of time, the evaluation is based on personal feelings and experience, without established standard pattern. OBJECTIVE: To present a guideline to evaluate the publications of scientific papers, without foreign influence. METHODS: The guideline to assess the value of each publication is based on the multiplication of three factors. The first factor is the value of the author corresponding to his place among the authors in the authorship of the work. The second factor is the classification of the type of publication. Finally, the product of these two factors is multiplied by ten times the "impact factor" of the journal. When the "impact factor" is zero, 0.1 should substitute it. RESULTS: This method has been used by us since 1994, with good results and without any problem related to the judgment. CONCLUSION: This criterion prevents subjective interference. Conflicts of interest are avoided when clearly presented from the beginning of the judgment process. PMID- 12886396 TI - [Cause of late death in liver transplant recipients]. AB - BACKGROUND: The objective is to present the causes of late death in patients subjected to liver transplantation. METHODS: A total of 209 patients were subjected to 223 liver transplantations (14 retransplantations). The computerized study protocol sheets were evaluated to determine the causes of late death (> 6 months after transplantation). RESULTS: Of the 209 patients, 30 had late death. Ductopenic rejection (chronic rejection) was the most common cause and it was observed in 10 patients. Time after transplantation at the moment of death of this group of patients varied from 11 to 57 months, with an average of 29 months. Seven patients died at the hospital admission of hepatic retransplantation. Other causes of late death were sepsis, lymphoproliferative disease, chronic renal insufficiency, and hepatic insufficiency. CONCLUSION: The most common cause of late death after liver transplantation is ductopenic rejection, followed by complications of retransplantation and sepsis. Death owing to ductopenic rejection may occur even many years after transplantation. PMID- 12886397 TI - [Dietetic and anthropometric assessment in juvenile rheumatoid arthritis patients]. AB - OBJECTIVES: to evaluate the dietary intake and the anthropometry of children and adolescents with juvenile rheumatoid arthritis - JRA, considering the pauciarticular and the polyarticular types. METHODS: Evaluation of the dietary intake of calories and macronutrients by the Food Register method compared with the Recommended Dietary Allowances - RDA, and the anthropometric evaluation by the z score of the height-for-age relationship and body mass index. RESULTS: The pauciarticular or polyarticular type did not determine any difference in calories, carbohydrates, proteins, and lipids intake, being the total caloric value, in both groups, bellow recommendations. Low-stature was observed in patients of the 2 groups, while leanness and obesity status were found in patients of the polyarticular group. CONCLUSIONS: JRA patients, particularly of the polyarticular group, can present compromised nutritional status and growth, probably owing to an inadequate dietary intake and increased need of calories and specific nutrients, which occur during the periods of the disease's activity. PMID- 12886398 TI - [An analyze the opportunities of early detection of breast cancer]. AB - PURPOSE: The study tried to identify and to analyze the opportunities of early detection of breast cancer available for the women of Botucatu County. METHODS: A sample of 261 women, 30 year-old or more was interviewed by telephone. RESULTS: The age average was of 51 years; 49.45% of them studied at least 9 year; 80% received general orientations about breast cancer prevention and 86,9% specific orientations on self breast exam; 78.9% of the interviewers made the self breast exam, but only 27% made it correctly. The physician breast examination was made in 76.2% of the women and mammography in 35.6%; if physicians requested mammography exam and if it was available for all of them, 83.1% of the women would like to do it. CONCLUSION: The breast cancer prevalence and the mortality are higher in old women. This study showed that these women presented a smaller rate of opportunities for early detection, because they were less informed about the right periodicity of the self breast exam and they had less mammography and physician breast examination frequency than the younger. The school degree was also related, because the women with more years of study had better opportunities than the ones with less years of study. PMID- 12886399 TI - [Influence of socioeconomic and nutritional factors on the evolution to complications in children hospitalized with pneumonia]. AB - BACKGROUND: To study the relationship between socioeconomic factors, nutritional status, evolution and complications in children with pneumonia. METHODS: A prospective, descriptive study on 85 children hospitalized with pneumonia from April 1999 to November 2000. RESULTS: Complications occurred in 31.8% children; of these, 26.6% - respiratory insufficiency, 44% -pleural effusion, 22% atelectasis. A significant relationship was observed between complications and the female sex (p=0.03; Odds Ratio=2.99 Confidence Interval 95%=1.16 -7.72) but this was not the case with age (p=0.98), maternal age (p=0.24), maternal educational level (p=0.62) and per capita income (p=0.07). No relationship was found between nutritional status and complication evolution, as well as with the other parameters (z score for the Waterlow classification, weight/age, height/age, weight/height and body mass index -p>0.05). CONCLUSIONS: Risk for complications was 2.99 times higher in the female group; no significant difference was found between the other parameters and complication evolution in children hospitalized with pneumonia. PMID- 12886400 TI - [Antroduodenal motility and pH changes after partial gastrectomy including the pacemaker region of the dog's stomach]. AB - BACKGROUND: Functional alterations of antroduodenal motility are not well defined at present. The gastric pacemaker is located at the greater curvature, next to the fundus. PURPOSE: The aim was to evaluate the influence of the gastric pacemaker on the gastroduodenal motility in dogs. METHODS: The study was designed in three steps and performed with 20 dogs: first, construction of a cervical esophagostomy to measure the antroduodenal motility and gastric pH-metry as validation step. The manometry was performed by means of a six point electronic manometry probe and both manometry and pH-metry were recorded during 4 hours. Second, the stomach's pacemaker region and gastric reservoir were resected by videolaparoscopy. Third, the antroduodenal motility and gastric pH-metry were measured again. RESULTS: In the validation step, there was established a typical dogs' antroduodenal motility pattern, with a three-shaped interdigestive motility complex (IMC) (phases 1 to 3); the gastric pH-metry showed a baseline at pH 1,15. After partial gastric resection, overall there was an increase in contractions' frequency, an increase in phase 2 and a decrease in phase 3 participation on the IMC's. Regarding the gastric pH measurements, there was an increase in pH intervals 0-1 and 1-2 and a decrease in pH intervals 6-7; overall, there was a decrease in the stomach's pH and a decrease in the prandial time. CONCLUSION: The resection of the stomach's pacemaker region and gastric reservoir correlated with alterations in antroduodenal motility, maintaining however the typical IMC pattern. PMID- 12886401 TI - [Indications of coronary cineangiography in coronary heart disease]. AB - Recent advances in Medical Science and the thechnological improvements in the field of myocardial revascularisation, in surgical procedures and in percutaneous interventions, made attractive the initial option for invasive strategies in the management of coronary heart disease. For this reason, coronary arteriography is nowadays more often indicated. Some concepts in coronary heart disease have been reviewed, specially those related to acute coronary syndromes. Non-ST-segment elevation myocardial infarction (previously called non-Q wave myocardial infarction) and unstable angina are now considered "unstable acute coronary syndromes" and both have the same guidelines for management. The main indications for coronary arteriography as the first diagnostic tool are: 1) incapacitating angina, even in stable patients; 2) high-risk patients with unstable coronary syndromes (refractory angina, troponin elevation, new ST- segment deviations, cardiac failure and serious arrythmias); 3) patients with acute ST-elevation myocardial infarction that will be submitted to primary angioplasty or with hemodynamic instability or persistent ischemia. Low-risk patients (angina that promptly subsides after medication, no electrocardiographic or laboratorial changes or atypical chest pain) may be submitted to non-invasive testing for further risk stratification; if no ischemia is detected, coronary arteriography is not indicated and optimized medical treatment is perfectly admitted for a great number of patients. The indications of coronary arteriography for the diagnosis and prognosis of coronary heart disease are not well delimited in clinical practice, and this method is frequently used as the first tool in the investigation of chest pain, even when the characteristics of pain are not exactly those of angina. In this review, the authors discuss the main indications of coronary arteriography in the multiple clinical aspects of coronary heart disease and in the differential diagnosis of chest pain. PMID- 12886402 TI - [Ciclosporine A-induced gingival hyperplasia]. AB - The increasing use of cyclosporine A (CSA) in organ transplants and in the treatment of autoimmune diseases has increased the incidence of cyclosporine A related adverse effects, including gingival hyperplasia (GH). GH causes esthetic, speech, mastication and tooth growth problems in the affected patients. The prevalence of cyclosporine-induced GH varies in different studies and may be as high as 85%, depending on the diagnostic criteria. This review approaches etiological, histological and clinical issues, as well as the prevention and treatment of this important injury. PMID- 12886403 TI - [Prophylactic routine anastomotic drainage in elective colorectal surgery: systematic review and metanalysis]. AB - OBJECTIVE: To analyse the results of the comparison of the use or not of routine prophylactic drainage of anastomoses after elective colorectal surgery. The following hypothesis will be tested: the use of routine prophylactic drainage of anastomoses after elective colorectal surgery is unjustified. PMID- 12886405 TI - Impact factor, scientific production and quality of Brazilian medical journals. PMID- 12886404 TI - [Munchhausen syndrome: diagnosis and management]. AB - The authors review the literature on Munchhausen syndrome, and ilustrate the clinical features of the disorder with the description of a characteristic case. Diagnosis and differential diagnosis are discussed with regard to other somatoform disorders such as conversion disorder and somatization disorder as well as to malingering and schizophrenia. The awareness of general practitioners and surgeons regarding this syndrome may avoid the exposure of these patients to serious complications of unnecessary medical and surgical procedures. The management of Muchhausen syndrome is aggravated by the low compliance in these patients. Early diagnosis could to a considerable extent prevent the iatrogenic risks. The authors recommend that patients presenting the psychopathological features of a Munchhausen syndrome should be conservatively observed and an attempt to clarify both the medical and the psychiatric diagnosis should be made before any invasive procedure is undertaken. PMID- 12886406 TI - Trypanosoma cruzi-elicited CD8+ T cell-mediated myocarditis: chemokine receptors and adhesion molecules as potential therapeutic targets to control chronic inflammation? AB - In Chagas disease, during the acute phase, the establishment of inflammatory processes is crucial for Trypanosoma cruzi control in target tissues and for the establishment of host/parasite equilibrium. However, in about 30% of the patients, inflammation becomes progressive, resulting in chronic disease, mainly characterized by myocarditis. Although several hypothesis have been raised to explain the pathogenesis of chagasic myocardiopathy, including the persistence of the parasite and/or participation of autoimmune processes, the molecular mechanisms underlying the establishment of the inflammatory process leading to parasitism control but also contributing to the maintenance of T. cruzi-elicited chronic myocarditis remain unsolved. Trying to shed light on these questions, we have for several years been working with murine models for Chagas disease that reproduce the acute self-resolving meningoencephalitis, the encephalitis resulting of reactivation described in immunodeficient individuals, and several aspects of the acute and chronic myocarditis. In the present review, our results are summarized and discussed under the light of the current literature. Furthermore, rational therapeutic intervention strategies based on integrin mediated adhesion and chemokine receptor-driven recruitment of leukocytes are proposed to control T. cruzi-elicited unbalanced inflammation. PMID- 12886407 TI - Epidemiology of Chagas disease in Guatemala: infection rate of Triatoma dimidiata, Triatoma nitida and Rhodnius prolixus (Hemiptera, Reduviidae) with Trypanosoma cruzi and Trypanosoma rangeli (Kinetoplastida, Trypanosomatidae). AB - A five-year domiciliary collection in the 22 departments of Guatemala showed that out of 4,128 triatomines collected, 1,675 were Triatoma dimidiata (Latreille, 1811), 2,344 were Rhodnius prolixus Stal 1859, and only 109 were T. nitida Usinger 1939. The Chagas disease parasite, Trypanosoma cruzi, was found in all three species. Their natural infection rates were similar in the first two species (20.6%; 19.1%) and slightly lower in T. nitida(13.8%). However there was no significant difference in the infection rates in the three species (p = 0.131). T. dimidiata males have higher infection rates than females (p = 0.030), whereas for R. prolixus there is no difference in infection rates between males and females (p = 0.114). The sex ratios for all three species were significantly skewed. More males than females were found inside houses for T. dimidiata (p < 0.0001) and T. nitida (p = 0.011); a different pattern was seen for R. prolixus (p = 0.037) where more females were found. Sex ratio is proposed as an index to show the mobility of T. dimidiata in different populations. T. dimidiata is widely distributed in the country, and is also the main vector in at least ten departments, but R. prolixus with higher vectorial capacity is an important vector in at least two departments. PMID- 12886408 TI - Coexistence of antibodies to tick-borne agents of babesiosis and Lyme borreliosis in patients from Cotia county, State of Sao Paulo, Brazil. AB - This paper reports a case of coinfection caused by pathogens of Lyme disease and babesiosis in brothers. This was the first case of borreliosis in Brazil, acquired in Cotia County, State of S o Paulo, Brazil. Both children had tick bite history, presented erythema migrans, fever, arthralgia, mialgia, and developed positive serology (ELISA and Western-blotting) directed to Borrelia burgdorferi G 39/40 and Babesia bovis antigens, mainly of IgM class antibodies, suggestive of acute disease. Also, high frequencies of antibodies to B. bovis was observed in a group of 59 Brazilian patients with Lyme borreliosis (25.4%), when compared with that obtained in a normal control group (10.2%) (chi-square = 5.6; p < 0.05). Interestingly, both children presented the highest titers for IgM antibodies directed to both infective diseases, among all patients with Lyme borreliosis. PMID- 12886409 TI - Isolation and identification of mycobacteria from livestock specimens and milk obtained in Brazil. AB - The prevalence of Mycobacterium bovis and other mycobacterial species in livestock specimens and milk was evaluated. An emphasis was placed upon the distribution of these organisms in milk that is readily available to the public that was either untreated, pasteurized, or treated using ultra high temperature. Twenty-two pathologic specimens from livestock (bovine, swine and bubaline) in five Brazilian states and 128 bovine milk samples from retail markets in the State of S o Paulo were examined for mycobacteria. Identification was made by classical biochemical tests, thin layer chromatography of mycolic acids and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis. Mycobacteria were isolated from 15 (68.2%) caseous lesions and from 23 (18%) milk samples. Eleven isolates were identified as M. bovis, and the remaining 27 nontuberculous mycobacterial isolates were represented by five species and six unidentified rapidly growing mycobacterial strains. The data demonstrate that animal products in Brazil are frequent reservoirs of mycobacteria and may pose a risk to the public. PMID- 12886410 TI - Determinants of human immunodeficiency virus (HIV) prevalence in homosexual and bisexual men screened for admission to a cohort study of HIV negatives in Belo Horizonte, Brazil: Project Horizonte. AB - Project Horizonte, an open cohort of homosexual and bisexual human immunodeficiency virus (HIV-1) negative men, is a component of the AIDS Vaccine Program, in Belo Horizonte, Minas Gerais, Brazil. The objective of this study was to compare volunteers testing HIV positive at cohort entry with a sample of those who tested HIV negative in order to identify risk factors for prevalent HIV infection, in a population being screened for enrollment at Project Horizonte. A nested case-control study was conducted. HIV positive volunteers at entry (cases) were matched by age and admission date to three HIV negative controls each. Selected variables used for the current analysis included demographic factors, sexual behavior and other risk factors for HIV infection. During the study period (1994-2001), among the 621 volunteers screened, 61 tested positive for HIV. Cases were matched to 183 HIV negative control subjects. After adjustments, the main risk factors associated with HIV infection were unprotected sex with an occasional partners, OR = 3.7 (CI 95% 1.3-10.6), receptive anal intercourse with an occasional partner, OR = 2.8 (95% CI 0.9-8.9) and belonging to the negro racial group, OR = 3.4 (CI 95% 1.1-11.9). These variables were associated with an increase in the risk of HIV infection among men who have sex with men at the screening for admission to an open HIV negative cohort. PMID- 12886411 TI - Comparative immunological recognition of proteins from New Guinea "C" dengue virus type 2 prototype and from a dengue virus type 2 strain isolated in the State of Rio de Janeiro, Brazil. AB - The protein profiles of the New Guinea "C" dengue virus type 2 (DENV-2)prototype and those of a Brazilian DENV-2 isolated in the State of Rio de Janeiro in 1995 were compared. SDS-PAGE analysis showed that the virus from Rio de Janeiro expresses NS5 (93.0 kDa), NS3 (66.8 kDa) E (62.4 kDa) and NS1 (41.2 kDa) proteins differently from the New Guinea "C" virus. The immunoblot revealed specificity and antigenicity for the NS3 protein from DENV-2 Rio de Janeiro mainly in primary infections, convalescent cases, and in secondary infections in both cases and only antigenicity for E and NS1 proteins for both viruses in primary and secondary infections. PMID- 12886412 TI - Comparative study of the stridulatory sulcus, buccula and rostrum of the nymphs of Triatoma guazu Lent & Wygodzinsky, 1979 and Triatoma jurbergi Carcavallo, Galvao & Lent, 1998 by scanning electron microscopy (Hemiptera, Reduviidae). AB - The ultrastructural morphology of the ventral region of the head (rostrum and buccula) and proesternum (stridulatory sulcus) of nymphs from the 1st to 5th instars of Triatoma guazu Lent & Wygodzinsky, 1979 and Triatoma jurbergi Carcavallo, Galvao & Lent, 1998 was described. Morphological differences between the two species and of the five nymphal stages development of each species were observed. These structures showed systematic differential characteristics of the studied species and may be used to increase their taxonomic range. PMID- 12886413 TI - Two new species of Nomimoscolex (Cestoda: Proteocephalidea, Monticelliidae) from Gymnotus carapo (Pisces: Gymnotiformes) in Argentina. AB - Nomimoscolex guillermoi n. sp. and N. dechambrieri n. sp. are described from the gymnotiform fish Gymnotus carapo from Argentina. The new species are placed into Nomimoscolex based on the cortical position of the vitelline follicles, and medullary position of the testes, ovary, and uterus. Both species were compared to the 13 species considered valid in the genus. The combination of features distinguishing N. guillermoi from N. dechambrieri is (1) the position of the vagina to cirrus pouch (anterior or posterior vs always anterior respectively), (2) the total number of testes (41-85 vs 108-130 respectively), (3) the distribution of the vitelline follicles (arranged in dorso-lateral and ventro lateral bands vs lateral bands respectively), (4) the length of the uteroduct (ending 58% vs 35% from posterior margin of mature proglottis respectively), and (5) the presence of gland cells in the scolex (unicellular glands in the apical region and the external margin of suckers vs the presence of unicellular glands in the apex and other grouped in a cluster medially to the suckers respectively). PMID- 12886414 TI - Description of Simulium damascenoi (Diptera: Simuliidae) male and the black-fly species from the State of Amapa, Brazil. AB - Five species are included in the Simulium siolii group, which is placed in the subgenus Psaroniocompsa (Diptera: Simuliidae). Of these five species, only two (Simulium siolii Py-Daniel and Simulium tergospinosum Hamada) have been described in all their life stages, except eggs. Knowledge of the taxonomic characters of all life stages of a species is important in order to clarify interspecific and higher-level taxonomic relationships. The objectives of the present study are to describe the male of Simulium damascenoi Py-Daniel, to provide a list of black fly species their bionomics and distributions in the state of Amap , Brazil, and to provide an identification key for larvae and pupae for these species. PMID- 12886415 TI - Reproductive system abnormalities in Schistosoma mansoni adult worms isolated from Nectomys squamipes (Muridae: Sigmodontinae): brightfield and confocal laser scanning microscopy analysis. AB - Schistosoma mansoni adult worms with genital anomalies isolated from Nectomys squamipes (Muridae: Sigmodontinae) were studied by confocal laser scanning microscopy under the reflected mode. One male without testicular lobes (testicular agenesia/anorchism) and two females, one with an atrophied ovary and another with 17 uterine eggs, were identified. The absence of testicular lobes occurred in a worm presenting otherwise normal male adult characteristics: tegument, tubercles and a gynaecophoric canal with spines. In both female specimens the digestive tube showed a vacuolated appearance, and the specimen with supernumerary uterine eggs exhibited a developing miracidium and an egg with a formed shell. The area of the ventral sucker was similar in both specimens however the tegument thickness, ovary and vitelline glands of the specimen with the atrophied ovary were smaller than those of the one with supernumerary eggs. These reported anomalies in the reproductive system call attention to the need to improve our understanding of genetic regulation and the possible role of environmental influences upon trematode development. PMID- 12886416 TI - Triatoma rubrovaria (Blanchard, 1843) (Hemiptera-Reduviidae-Triatominae) III: patterns of feeding, defecation and resistance to starvation. AB - Data from the Chagas Disease Control Program indicate a growing domiciliary and peridomiciliary invasion of Triatoma rubrovaria in the State of Rio Grande do Sul, where it has become the most frequent triatomine species captured there since the control of T. infestans. Bionomic characteristics that could influence the vectorial capacity of T. rubrovaria as vector of Trypanosoma cruzi were evaluated: patterns of (i) feeding, (ii) defecation, and (iii) resistance to starvation, using insects fed on mice. Fifty three percent of the females showed a defecation pattern conducive to chagasic transmission, defecating either on or near the bite site. The averages of the resistance to starvation varied from 48.1 to 179 days, for the first and fifth nymphal stages, respectively. Our study shows that with respect to the patterns of feeding, defecation and resistance to fasting, T. rubrovaria presented similar rates to the ones observed for other effective vector species, such as T. infestans. Thus, based on our studies we conclude that T. rubrovaria has biological characteristics that can positively influence its capacity to become infected and transmit T. cruzi, and also to keep residual populations after chemical control interventions. PMID- 12886417 TI - Detection of Trypanosoma cruzi DNA within murine cardiac tissue sections by in situ polymerase chain reaction. AB - The use of in situ techniques to detect DNA and RNA sequences has proven to be an invaluable technique with paraffin-embedded tissue. Advances in non-radioactive detection systems have further made these procedures shorter and safer. We report the detection of Trypanosoma cruzi, the causative agent of Chagas disease, via indirect and direct in situ polymerace chain reaction within paraffin-embedded murine cardiac tissue sections. The presence of three T. cruzi specific DNA sequences were evaluated: a 122 base pair (bp) sequence localized within the minicircle network, a 188 bp satellite nuclear repetitive sequence and a 177 bp sequence that codes for a flagellar protein. In situ hybridization alone was sensitive enough to detect all three T. cruzi specific DNA sequences. PMID- 12886418 TI - The 245 kb amplified chromosome of Leishmania (V.) braziliensis contains a biopterin transporter gene. AB - Leishmania (V.) braziliensis M2903 presents a small linear and stable 245 kb chromosome originating from a genomic amplification. Similar amplifications present in other species of Leishmania contain a gene coding for a biopterin transporter. Since Leishmania is auxotrophic for this metabolite, this amplification could result from the need to better capture biotpterin from growth media under specific circumstances. In this paper we show that this gene is also present in L. (V.) braziliensis small chromosome, which shares sequences with other genomic amplifications already described. PMID- 12886419 TI - Brazilian Flavivirus phylogeny based on NS5. AB - In this work, a comprehensive phylogenetic study based on 600 base pair nucleotide and on putative 200 amino acid sequences of NS5 was carried out in order to establish genetic relationships among 15 strains of 10 Brazilian flaviviruses: Bussuquara, Cacipacore, dengue type 1, 2 and 4, Iguape, Ilheus, Rocio, Saint Louis encephalitis (SLE), and yellow fever. Phylogenetic trees were created by neighbor-joining and maximum parsimony methods. These trees showed Brazilian flaviviruses grouped into three main branches: yellow fever branch, dengue branch subdivided in types 1, 2 and 4 branches, and Japanese encephalitis virus (JEV) complex branch including SLE virus strains, Cacipacore, Iguape, Rocio, Ilheus and Bussuquara. Viruses transmitted by Aedes mosquitoes, such as dengue and urban yellow fever, that are also the only Flavivirus causing hemorrhagic fevers in Brazil, were grouped in the same cluster. Encephalitis associated viruses, transmitted by Culex mosquitoes such as JEV complex branch including SLE virus strains, Cacipacore, Iguape, Rocio, Ilheus and Bussuquara were also grouped in the same clade. PMID- 12886420 TI - Malpighian tubule polytene chromosomes of Culex quinquefasciatus (Diptera, Culicinae). AB - Dipteran polytene chromosomes provide an excellent model for understanding in species complexes, as well as for structural and functional cytogenetics. The status of species in the Culex pipiens complex is controversial and the use of polytene chromosomes for cytogenetic analysis in the subfamily Culicinae has been difficult because of methodological problems. In this study, Malpighian tubule polytene chromosomes were obtained from young (0 to 12 h, 20 C) and old (20 to 42 h, 28 C) laboratory-bred C. pipiens quinquefasciatus pupae. The chromosome maps for this species were constructed and compared with published data for C. pipiens pipiens and C. p. quinquefasciatus. Although the banding patterns were conserved between subspecies, analysis of the structural variations in the bands and interbands revealed differences apparently related to the physiological stage and ecogeographical strain. The organization of the centromeric regions in larval and pupal chromosomes showed greater similarity to each other than did those of pupal and adult chromosomes. The use of pupal polytene chromosomes for in situ hybridization with vector competence probes is discussed. PMID- 12886421 TI - A technique for preparing polytene chromosomes from Aedes aegypti (Diptera, Culicinae). AB - Polytene chromosome preparations were obtained from larval, pupal and adult female Malpighian tubules of Aedes aegypti. The Malpighian tubules of the pupae (0-4 h old) from larvae reared at 20 C provided the best cytogenetic analysis. The interaction of nucleic acids and proteins that influence the spreading of the chromosomes could be reduced with the preparation technique of the sheets submitted to a stronger treatment starting with the hypotony of tissue and successive bathings with acetic acid. A simple technique should facilitate molecular cytogenetics used in the location of resistance and vector competence genes. PMID- 12886422 TI - The use of polysiloxane/polyvinyl alcohol beads as solid phase in IgG anti Toxocara canis detection using a recombinant antigen. AB - Immunodetection of human IgG anti-Toxocara canis was developed based on ELISA and on the use of polysiloxane/polyvinyl alcohol (POS/PVA) beads. A recombinant antigen was covalently immobilized, via glutaraldehyde, onto this hybrid inorganic-organic composite, which was prepared by the sol-gel technique. Using only 31.2 ng antigen per bead, a peroxidase conjugate dilution of 1:10,000 and a serum dilution of 1:200 were adequate for the establishment of the procedure. This procedure is comparable to that which utilizes the adsorption of the antigen to conventional PVC plates. However, the difference between positive and negative sera mean absorbances was larger for this new glass based assay. In addition to the performance of the POS/PVA bead as a matrix for immunodetection, its easy synthesis and low cost are additional advantages for commercial application. PMID- 12886423 TI - Antigenic characterization of Brazilian isolates of Anaplasma marginale. AB - Antigenic characterization of Anaplasma marginale isolates, by identifying conserved and variable epitopes of major surface proteins (MSP), is an important tool for vaccine development against this rickettsia. The B cell epitopes of A. marginale isolates from three microregions of the State of Pernambuco and one from the State of Mato Grosso do Sul, Brazil, were characterized by indirect fluorescent antibody technique (IFAT) and Western blot (WB) with 15 monoclonal antibodies (MAbs). The epitope recognized by MAb ANA22B1 (MSP-1a) was conserved by IFAT and WB (73-81 kDa). MSP-2 epitopes recognized by MAbs ANAO58A2 and ANAO70A2 were conserved by IFAT, while ANAO50A2 and ANA66A2 epitopes were polymorphic; in the WB, the MAbs ANAO50A2 and ANAO70A2 identified bands of 45 kDa only in the Pernambuco-Mata isolate. None of the isolates reacted with MAb ANAR75C2 (MSP-3). The MSP-4 epitope recognized by MAb ANAR76A1 was conserved by IFAT, as well as the MSP-5 epitope recognized by MAb ANAF16C1 by IFAT and WB (16 kDa). The MAbs ANAR17A6, ANAR83B3, ANAR94C1, ANAO24D5 and ANAR19A6 identified conserved epitopes by IFAT. MSP-1, MSP-2 and MSP-4, which previously showed partial protection in experimental trials, are also potential immunogens to be employed in Brazil, due to the B cell epitope conservation. PMID- 12886424 TI - Species distribution and antifungal susceptibility profile of Candida spp. bloodstream isolates from Latin American hospitals. AB - From March 1999 to March 2000, we conducted a prospective multicenter study of candidemia involving five tertiary care hospitals from four countries in Latin America. Yeast isolates were identified by classical methods and the antifungal susceptibility profile was determined according to the National Committee for Clinical Laboratory Standards microbroth assay method. During a 12 month-period we were able to collect a total of 103 bloodstream isolates of Candida spp. C. albicans was the most frequently isolated species accounting for 42% of all isolates. Non-albicans Candida species strains accounted for 58% of all episodes of candidemia and were mostly represented by C. tropicalis (24.2%) and C. parapsilosis (21.3%). It is noteworthy that we were able to identify two cases of C. lusitaniae from different institutions. In our casuistic, non-albicans Candida species isolates related to candidemic episodes were susceptible to fluconazole. Continuously surveillance programs are needed in order to identify possible changes in the species distribution and antifungal susceptibility patterns of yeasts that may occurs after increasing the use of azoles in Latin American hospitals. PMID- 12886425 TI - Increased plasma levels of tumor necrosis factor-alpha in asymptomatic/"indeterminate" and Chagas disease cardiomyopathy patients. AB - We compared plasma tumor necrosis factor-alpha (TNF-alpha) levels among asymptomatic/"indeterminate" Chagas disease patients (ASY) and patients across the clinical spectrum of chronic Chagas disease cardiomyopathy (CCC). Idiopathic dilated cardiomyopathy (DCM) patients and normal controls (NC) were included as controls. ASY Chagas disease patients had significantly higher plasma TNF-alpha levels than NC. TNF-alpha levels among severe CCC patients with significant left ventricular (LV) dysfunction were similar to those of DCM patients, showing average 2-fold higher levels than CCC patients without LV dysfunction and ASY patients, and 8-fold higher levels than NC. In Chagas disease, chronic TNF-a production prior to heart failure may play a role in CCC progression. PMID- 12886426 TI - Diterpenoids from Azorella compacta (Umbelliferae) active on Trypanosoma cruzi. AB - The anti-Trypanosoma cruzi activity of natural products isolated from Azorella compacta was evaluated, with particular emphasis on their effect against intracellular amastigotes. Five diterpenoids from A. compacta derived from mulinane and azorellane were isolated and identified. Only two products, named azorellanol (Y-2) and mulin-11,3-dien-20-oic acid (Y-5), showed trypanocidal activity against all stages of T. cruzi including intracellular amastigotes. At 10 M, these compounds displayed a strong lytic activity. It ranged from 88.4 0.6 to 99.0 1 % for all strains and stages evaluate, with an IC50 /18 h values of 20-84 M and 41-87 M, respectively. The development of intracellular amastigotes was also inhibited by nearly 60% at 25 M. The trypanocidal molecules Y-2 and Y-5 did show different degrees of cytotoxicity depending on the cell line tested, with an IC50 /24 h ranging from 33.2 to 161.2 M. We evaluated the effect of diterpenoids against intracellular T. cruzi forms by immunofluorescent identification of a specific membrane molecular marker (Ssp-4 antigen) of the T. cruzi amastigote forms. The accuracy and reproducibility of the measurements were found to be outstanding when examined by confocal microscopy. PMID- 12886427 TI - The effect of nitric oxide combined with fluoroquinolones against Salmonella enterica serovar Typhimurium in vitro. AB - Two regulons, soxRS and marRAB, are associated with resistance to quinolones or multiple antibiotic in Salmonella enterica serovar Typhimurium. These regulons are activated by nitric oxide and redox-cycling drugs, such as paraquat and cause on activation of the acrAB-encoded efflux pump. In this study, we investigated the effect of nitric oxide (NO) alone and in combination with ofloxacin, ciprofloxacin, and pefloxacin against S. typhimurium clinical isolates and mutant strains in vitro. We did not observe synergistic effect against clinical isolates and SH5014 (parent strain of acr mutant), while we found synergistic effect against PP120 (soxRS mutant) and SH7616 (an acr mutant) S. typhimurium for all quinolones. Our results suggest that the efficiencies of some antibiotics, including ofloxacin, ciprofloxacin, and pefloxacin are decreased via activation of soxRS and marRAB regulons by NO in S. enterica serovar Typhimurium. Further studies are warranted to establish the interaction of NO with the genes of Salmonella and, with multiple antibiotic resistance. PMID- 12886428 TI - Molluscicidal activity of Physalis angulata L. extracts and fractions on Biomphalaria tenagophila (d'Orbigny, 1835) under laboratory conditions. AB - The main objective of this research is to evaluate the molluscicide activity of Physalis angulata L. Biomphalaria tenagophila specimens under laboratory conditions. Extracts and fractions were supplied by the Laborat rio de Qu mica de Produtos Naturais, Farmanguinhos-Fiocruz. Experiments were performed according to the methodology described by the World Health Organization for molluscicide tests using the concentrations from 0.1 to 500 mg/l of the extracts, fractions and of a pool of physalins modified steroids present in this species. The results show that ethyl acetate and acetone extracts from the whole plant, the ethanolic extracts of the roots and the physalins pool from stems and leaves were active. Only the whole plant extracts were available in sufficient quantity for the determination of LD50 and LD90 values. PMID- 12886429 TI - Apparent competition through facilitation between Melanoides tuberculata and Biomphalaria glabrata and the control of schistosomiasis. AB - Interactions between two species that result in reduced growth rates for both and extinction of one of the species are generally considered cases of asymmetric interspecific competition. Exploitative or interference competition is the usual mechanism invoked. Here we describe another mechanism producing the same result, named apparent competition through facilitation (ACF), observed between Melanoides tuberculata and Biomphalaria glabrata populations. The superior competitor actually gives some benefit to the other species, whose population becomes unstable with progressively increasing oscillations, leading to extinction. A model of ACF using difference equations suggests initial dynamics distinct from traditional interspecific competition. The dynamics of two freshwater snails in the field and in laboratory experiments suggest ACF, and these relations should be considered in studies of schistosomiasis control. ACF could occur in natural populations, but might have gone undetected because the final result is similar to traditional interspecific competition. PMID- 12886430 TI - [The nutritional transition and the epidemiology of obesity in Latin America]. PMID- 12886431 TI - [Treatment of obesity: the need to target attention on high-risk patients characterized by abdominal obesity]. AB - Abdominal obesity is associated with metabolic abnormalities, increasing the risk of type 2 diabetes and coronary artery disease (CAD). The Quebec Cardiovascular Survey demonstrated that the atherogenic metabolic triad (AMT) present in abdominally obese (AO) males increases the risk of CAD 20-fold over the course of 5 years. An early detection algorithm was developed to identify individuals presenting these atherogenic abnormalities. It was found that the association of large waist circumference (WC) and moderate hypertriglyceridemia (the "hypertriglyceridemic waist", or HW) could adequately identify a significant portion of individuals with the AMT. It is important to note that even in the absence of classic risk factors, abdominally obese patients can present increased risk of CAD if they have HW. Finally, it has been suggested that the risk of developing an acute coronary syndrome in AO patients is not always related to the degree of coronary stenosis, and the patient s atherothrombotic/inflammatory profile should be taken into account in evaluating risk. Stabilization of the atherosclerotic plaque would become a legitimate therapeutic objective, and more feasible for prevention of CAD, in AO patients. PMID- 12886432 TI - Stunting and future risk of obesity: principal physiological mechanisms. AB - There is a fair amount of epidemiological evidence showing that nutritional stunting causes increased risks of obesity. Obesity is increasing dramatically not only in developed countries but also in developing countries, such as Brazil, especially among the poorer. The mere coexistence of undernutrition and obesity among poor people has a great impact, as the burden in the social, economic, and health care systems is remarkable. In addition, an increasing number of studies have shown that nutritional stunting causes a series of important long-lasting changes such as lower energy expenditure, higher susceptibility to the effects of high-fat diets, lower fat oxidation, and impaired regulation of food intake. These findings suggest that a broader and more detailed understanding of the long lasting effects of early undernutrition, direct cause of nutritional stunting, is needed. Within this context, we present data of some physiological mechanisms that substantiate the association between previous undernutrition and future obesity. PMID- 12886433 TI - [Trends in diabetes mellitus in Brazil: the role of the nutritional transition]. AB - Prevalence of type 2 diabetes mellitus has increased rapidly and is expected to increase even further. In Latin America countries there is an upward trend among the younger age groups, with a major negative impact on the quality of life and burden of the disease for the health care system. Increasing rates of overweight and obesity associated with lifestyle changes and aging of the population are the main contributing factors to the increasing prevalence of type 2 diabetes. Dietary shifts in the Brazilian population (low frequency of fiber-rich foods and heavy consumption of saturated fatty acids and sugars) and sedentary lifestyle are the main risk factors for obesity, type 2 diabetes, and other chronic diseases. Diabetes prevention programs are being promoted in several countries with results showing improvement in quality of life. PMID- 12886434 TI - [A review of breastfeeding in Brazil and how the country has reached ten months' breastfeeding duration]. AB - In 1975, one out of two Brazilian women only breastfed until the second or third month; in a survey from 1999, one out of two breastfed for 10 months. This increase over the course of 25 years can be viewed as a success, but it also shows that many activities could be better organized, coordinated, and corrected when errors occur. Various relevant decisions have been made by international health agencies during this period, in addition to studies on breastfeeding that have reoriented practice. We propose to review the history of the Brazilian national program to promote breastfeeding, focusing on an analysis of the influence of international policies and analyzing them in four periods: 1975-1981 (when little was done), 1981-1986 (media campaigns), 1986-1996 (breastfeeding friendly policies), and 1996-2002 (planning and human resources training activities backed by policies to protect breastfeeding). The challenge for the future is to continue to promote exclusive breastfeeding until the sixth month, taking specific population groups into account. PMID- 12886435 TI - [Factors associated with dietary patterns in the urban Brazilian population]. AB - This study evaluated factors associated with dietary patterns in the Brazilian population based on the Living Standards Survey conducted in the Northeast and Southeast regions of the country. Multi-stage probability sampling was employed to select the households, and for the present analysis 5,121 adults aged 20 to 50 years were included. Pregnant women and individuals reporting chronic health conditions were excluded. Through principal component analysis, two major family dietary patterns were identified: a mixed pattern, in which all groups and foods have approximately the same factor loading, and a second pattern based main1y on rice and beans, which was called a traditional diet. Weight and height were measured in the households, and food intake was based on a 21-item semi quantitative family questionnaire. The Northeast (as compared to the Southeast) was negatively associated with the mixed pattern. Body mass index was positively associated with the mixed pattern, whereas leisure physical activity and Black skin color were negatively associated with the mixed pattern. Schooling and income levels explained most of the dietary variance, but after adjusting for education and income, region of residence remained significantly associated and was the third most important explanatory variable. PMID- 12886436 TI - [Socio-demographic and lifestyle factors associated with abdominal fat distribution in adults: a population-based survey in Southern Brazil]. AB - This article describes the anthropometric distribution of abdominal fat in the urban adult population in Pelotas, Rio Grande do Sul, Brazil, and the influence of independent variables on this distribution. In a cross-sectional population based study, 3,464 adults from 20 to 69 years of age were selected in a multistage systematic sampling. They were interviewed and had their largest abdominal circumference measured. Means were compared using one-way ANOVA, while multiple linear regression models were employed to adjust for confounding. According to the study, older white married men with higher family incomes were more likely to have the highest mean abdominal circumferences (p < 0.001). Women at greatest risk for abdominal fat were older married mothers with a history of 4 or more pregnancies and less schooling (p < 0.001). Former smokers had the highest mean abdominal circumferences in both males and females, while other lifestyle factors such as alcohol consumption and physical activity showed no association with the outcome after adjusting for confounding. PMID- 12886437 TI - [The changing relationship between education and risk of obesity in Brazil (1975 1997)]. AB - Based on three comparable surveys conducted in the last quarter of the 20th century in the most heavily populated areas of Brazil, a substantial change was observed in the secular trend towards obesity based on levels of schooling in the population. While during the first period (1975-1989) there was an increasing risk of obesity for all levels of schooling, with the greatest upward trend for men and women with more schooling, during the second period (1989-1997) the increase in obesity was the greatest for individuals with no schooling, along with a stabilization or even decrease in this condition among groups of females with medium or higher education. As a result of the upward trend, the positive relationship decreased between schooling and risk of obesity in men, and the inverse relationship already observed in the female population was further increased. The main potential implications of these findings are for public health. PMID- 12886438 TI - Obesity trends and determinant factors in Latin America. AB - Obesity rates have increased markedly in Latin America, especially during the last 10-15 years, becoming a public health problem in most countries. Prevalence of obesity among preschool children remains low, while among schoolchildren it has increased considerably. Prevalence is high in the adult population, especially among women with less schooling. In developed populations, obesity occurs more frequently among the poor; the opposite occurs in less developed societies, where in households undergoing nutritional transition, underweight can coexist with obesity. The most important determinant factors involved in the increasing obesity prevalence are fetal and infant nutritional conditions (stunting), education and socioeconomic conditions, dietary changes (especially increased total energy intake), and physical inactivity. Because chronic diseases are the main causes of death in the Region and obesity is one of the main risk factors for these diseases, policies to improve economic and educational levels with the implementation of health promotion and prevention should be a priority in every country. PMID- 12886439 TI - Trends in dietary patterns of Latin American populations. AB - It is important to characterize the level and magnitude of changes in food consumption patterns in Latin American populations as they undergo demographic and developmental transitions because of the effects of such changes on the development and progression of chronic diseases. This paper examines trends in food intake across regions in Latin America. Although trends in apparent food consumption differ in magnitude and timing, the overall patterns of change are remarkably consistent. Intakes of total fat, animal products, and sugar are increasing, even while there have been rapid declines in the intake of cereals, fruit, and some vegetables. The costs of the increased prevalence of chronic disease associated with these dietary changes are already affecting health systems still coping with malnutrition and infectious disease. Because this pattern of change is predictable, it is important to learn from the experiences gained in countries that are more advanced in the transition. Efforts to educate the population on the importance of a healthy diet and to issue policies to improve the availability of a healthy food supply can help to reduce the rapid escalation of obesity and chronic diseases. PMID- 12886440 TI - [The obesity epidemic and related factors: the case of Spain]. AB - The obesity epidemic is a global phenomenon that does not respect geographic or socio-demographic boundaries. Thus, research on factors related to the obesity epidemic has focused on social and economic characteristics of modern societies. This article discusses obesity in Spain and trends in several associated factors. Together with the increase in the prevalence of obesity, important changes in the population's dietary pattern have been observed, although total energy and fat intake appear to be stable. According to several indirect indicators, sedentary behavior predominates, although the percentage of the population reporting some leisure exercise is increasing. An increase in the amount of leisure time in modern societies is suggested as an explanation for this paradox. Factors related to energy expenditure could be as important as dietary factors in the genesis of the obesity epidemic. PMID- 12886441 TI - [Menarche, early pregnancy, and obesity in selected Brazilian women from a health care center in Belo Horizonte, Minas Gerais, Brazil]. AB - The aim of this study was to investigate obstetric variables potentially associated with obesity among 486 Brazilian childbearing-age women aged 15-59 residing in the municipality of Belo Horizonte, Minas Gerais State. Body fat (BF) was measured through impedance analysis, and obesity was defined as BF > 30%. The association between obstetrics factors and obesity was evaluated through multivariate logistic regression. The following variables remained in the logistic regression after adjustments for total income, smoking, alcohol consumption, and physical activity: age at menarche < 12 years (OR = 3.02; 95% CI: 1.62-5.61), age > 30 and < 39 (OR = 1.72; 95% CI: 1.01-2.92), age > 40 years (OR = 3.32; 95% CI: 1.76-6.27), age at first childbirth (OR = 1.99; 95% CI: 1.07 3.68), and the following interaction: age group > 30 and < 39 and age at menarche (OR = 0.27; 95% CI: 0.09-0.83). It appears that obstetric factors affect obesity through a complex network of interrelations that involve the covariates studied above. It is important to support efficient programs to prevent obesity, as well as family planning programs emphasizing a reduction in the prevalence of teenage pregnancies. PMID- 12886442 TI - Breastfeeding and the nutritional transition in the Latin American and Caribbean Region: a success story? AB - The objectives of this paper are to examine recent breastfeeding duration trends in Latin America and the Caribbean to document: (a) rural-urban differentials, (b) differences in educational levels, and (c) changes in breastfeeding duration across time. Secondary data analyses were conducted with 23 Demographic and Health Surveys collected between the mid-1980s and mid-1990s. Results indicate that median breastfeeding duration is still greater in rural (as compared to urban) areas and among less (versus more) educated women, although these differentials are decreasing with time. In five of the six countries examined for secular trends, breastfeeding duration continues to increase in both rural and urban areas. Breastfeeding duration in urban and rural areas was strongly correlated within countries. Breastfeeding duration improved more among women with the highest and declined among those with the lowest levels of education. Results indicate that breastfeeding duration has increased in Latin America and the Caribbean at a time when the opposite was predicted, given the region's increased urbanization. Breastfeeding protection policies and promotion programs may explain part of the increase in breastfeeding duration. PMID- 12886443 TI - [Prevalence of overweight and obesity and associated factors among adolescents in the Northeast and Southeast regions of Brazil, 1996 to 1997]. AB - The purpose of this study was to assess the prevalence and factors associated with overweight and obesity (OW/O) among Brazilian adolescents, based on the Living Standards Survey conducted in 1996 and 1997 (IBGE, 1998a). The sample included 1,027 and 854 adolescents aged 15 to 19 years from the Northeast and Southeast regions, respectively. Body Mass Index (w/h2) was calculated. Statistical analysis considered expansion factors and sample design. Prevalence of OW/O was 8.45% in the Northeast and 11.53% in the Southeast. In the Northeast there was a higher risk of OW/O for girls (PR = 3.00), with the same situation in both urban (PR = 3.21) and rural areas (PR = 2.27). In the Southeast, the risk of OW/O was lower among girls (PR = 0.58). The same situation occurred in girls from urban areas (PR = 0.51), but in rural areas there was an increased risk among girls (PR = 1.86). Per capita monthly income was associated with the risk of OW/O among boys in both regions. Programs to prevent OW/O should incorporate nutritional reeducation and encourage physical exercise, besides considering socioeconomic, cultural, and regional differences and the location of the target population. PMID- 12886444 TI - [Trends in the nutritional profile of children born in 1993 in Pelotas, Rio Grande do Sul, Brazil: longitudinal analyses]. AB - Trends in nutritional status were studied in children born in 1993 in the city of Pelotas, Rio Grande do Sul State, Brazil. During that year, all infants born in the city were identified, and a sub-sample of 1,273 children was visited twice at home for anthropometric examinations at an average age of 12 and 54 months, respectively. Approximately 10% of all children were lost to follow up from birth to 4 years. The prevalence of nutritional deficit decreased, while that of overweight increased between ages 1 and 4 years. The prevalence of nutritional deficit showed an inverse association with birth weight, while overweight showed a direct relationship. About 50% of the children with height/age deficit at 12 months remained in this same condition at 4 years old, while a third of overweight children remained in this condition during the same period. On the other hand, some 10% of non-overweight children became overweight between 1 and 4 years of age. The nutritional transition observed in Pelotas requires a paradigm shift in health services, which are still concentrated on detecting and treating malnutrition rather than obesity. PMID- 12886445 TI - Postpartum weight retention among women in Rio de Janeiro: a follow-up study. AB - This study presents follow-up results on trends in postpartum weight retention among women in the city of Rio de Janeiro, Brazil, at 0.5, 2, 6, and 9 months postpartum. The outcome variable, weight retention, was calculated by subtracting the reported pre-pregnancy weight from the observed weight at each interview. Statistical analyses used means and a 95% confidence interval for weight retention. Mean weight retention was 4.7, 4.1, 3.4, and 3.1kg at 0.5, 2, 6 and 9 months postpartum, respectively. At completion of the study, the largest weight retention was observed in women over 30 years of age (4.2kg) and with 30% or more of body fat (5.9kg). The rate of reduction in weight retention was 0.4kg/month, 0.2kg/month, and 0.1kg/ month for the periods 0.5-2, 2-6, and 6-9 months, respectively. The largest reductions were observed among young, single women and those with < 30% of body fat at baseline. The results suggest that variables such as age, parity, schooling, and body fat may be important predictors of postpartum weight retention. PMID- 12886446 TI - [Obesity in Latin American children: situation, diagnostic criteria and challenges]. AB - Obesity has emerged as an alarming public health problem, having increased rapidly in both adults and children. Considering 12 countries with reported levels of overweight and obesity, one third have a 20% prevalence of one or both conditions. In 17 countries reporting data on obesity, the mean rate was 4.6%; four countries reported rates over 6%, while three reported less than 2%. The majority of these countries showed increases of 70% in overweight and 60% in obesity during the last decade. Only two countries reported decreasing prevalence rates. Countries may be classified in four groups: (1) low or even downward prevalence of overweight and obesity; (2) underweight reported simultaneously with an increase in obesity; (3) a large territory and population with differences in the prevalence of overweight and obesity between regions and socioeconomic groups; and (4) little underweight and high prevalence of obesity. In conclusion, there is no doubt about the increasing trend in overweight and obesity among Latin American children. However, since there is no consensus as to diagnostic criteria for overweight and obesity, the real prevalence is not known. PMID- 12886447 TI - [Growth and nutritional status in a probabilistic sample of schoolchildren from Rio de Janeiro, 1999]. AB - Growth and nutritional status were assessed in a probabilistic sample of public schoolchildren (1,705 girls and 1,682 boys) in the city of Rio de Janeiro in 1999. Nutritional status of the under-10-year group was assessed by z < -2 (undernutrition) for body mass for height and height for age and by z > +2 for body mass for height (overweight). For children over 10, the 5th and 85th percentiles of body mass index for age were used to assess underweight and overweight, respectively. Prevalence rates of overweight and obesity according to the International Obesity Task Force criteria were also calculated. Prevalence rates for undernutrition were below expected for the reference population. Prevalence of overweight was approximately 18% and 14% for girls and boys, respectively, and obesity was 5% for both sexes. The results of the present study are similar to findings from population-based surveys in Brazil: low prevalence of undernutrition and high prevalence of overnutrition. PMID- 12886448 TI - [Nutritional transition in Brazil: geographic and temporal trends]. AB - Based on three cross-sectional studies conducted in the 1970s, 80s, and 90s, an analysis of the nutritional transition in Brazil was performed. This analysis found a rapid decrease in the prevalence of childhood malnutrition and an even more rapid increase in adult overweight/obesity. Correction of the height deficit was 72% in urban children and 54.4% in rural children. Obesity rates doubled or tripled in adult men and women at the extremes of the time series analyzed. Excluding the rural Northeast, malnutrition prevalence in adult women dropped to acceptable rates (about 5%) starting in 1989. Inversely, anemia continued to display a high prevalence, and there is indication of an epidemic trend. The evolution of nutritional status in the Brazilian population according to macro regions and social distribution is reported. The likely factors for the observed changes are analyzed. PMID- 12886449 TI - Dimensions and dynamics in integrin function. AB - Integrins play crucial roles in cell adhesion, migration, and signaling by providing transmembrane links between the extracellular matrix and the cytoskeleton. Integrins cluster in macromolecular complexes to generate cell matrix adhesions such as focal adhesions. In this mini-review, we compare certain integrin-based biological responses and signaling during cell interactions with standard 2D cell culture versus 3D matrices. Besides responding to the composition of the matrix, cells sense and react to physical properties that include three-dimensionality and rigidity. In routine cell culture, fibroblasts and mesenchymal cells appear to use focal adhesions as anchors. They then use intracellular actomyosin contractility and dynamic, directional integrin movements to stretch cell-surface fibronectin and to generate characteristic long fibrils of fibronectin in "fibrillar adhesions". Some cells in culture proceed to produce dense, three-dimensional matrices similar to in vivo matrix, as opposed to the flat, rigid, two-dimensional surfaces habitually used for cell culture. Cells within such more natural 3D matrices form a distinctive class of adhesion termed "3D-matrix adhesions". These 3D adhesions show distinctive morphology and molecular composition. Their formation is heavily dependent on interactions between integrin alpha 5 beta 1 and fibronectin. Cells adhere much more rapidly to 3D matrices. They also show more rapid morphological changes, migration, and proliferation compared to most 2D matrices or 3D collagen gels. Particularly notable are low levels of tyrosine phosphorylation of focal adhesion kinase and moderate increases in activated mitogen-activated protein kinase. These findings underscore the importance of the dimensionality and dynamics of matrix substrates in cellular responses to the extracellular matrix. PMID- 12886450 TI - Importance of hyaluronan biosynthesis and degradation in cell differentiation and tumor formation. AB - Hyaluronan is an important connective tissue glycosaminoglycan. Elevated hyaluronan biosynthesis is a common feature during tissue remodeling under both physiological and pathological conditions. Through its interactions with hyaladherins, hyaluronan affects several cellular functions such as cell migration and differentiation. The activities of hyaluronan-synthesizing and degrading enzymes have been shown to be regulated in response to growth factors. During tumor progression hyaluronan stimulates tumor cell growth and invasiveness. Thus, elucidation of the molecular mechanisms which regulate the activities of hyaluronan-synthesizing and -degrading enzymes during tumor progression is highly desired. PMID- 12886451 TI - What can digital transcript profiling reveal about human cancers? AB - Important biological and clinical features of malignancy are reflected in its transcript pattern. Recent advances in gene expression technology and informatics have provided a powerful new means to obtain and interpret these expression patterns. A comprehensive approach to expression profiling is serial analysis of gene expression (SAGE), which provides digital information on transcript levels. SAGE works by counting transcripts and storing these digital values electronically, providing absolute gene expression levels that make historical comparisons possible. SAGE produces a comprehensive profile of gene expression and can be used to search for candidate tumor markers or antigens in a limited number of samples. The Cancer Genome Anatomy Project has created a SAGE database of human gene expression levels for many different tumors and normal reference tissues and provides online tools for viewing, comparing, and downloading expression profiles. Digital expression profiling using SAGE and informatics have been useful for identifying genes that have a role in tumor invasion and other aspects of tumor progression. PMID- 12886452 TI - Endothelial cells, tissue factor and infectious diseases. AB - Tissue factor is a transmembrane procoagulant glycoprotein and a member of the cytokine receptor superfamily. It activates the extrinsic coagulation pathway, and induces the formation of a fibrin clot. Tissue factor is important for both normal homeostasis and the development of many thrombotic diseases. A wide variety of cells are able to synthesize and express tissue factor, including monocytes, granulocytes, platelets and endothelial cells. Tissue factor expression can be induced by cell surface components of pathogenic microorganisms, proinflammatory cytokines and membrane microparticles released from activated host cells. Tissue factor plays an important role in initiating thrombosis associated with inflammation during infection, sepsis, and organ transplant rejection. Recent findings suggest that tissue factor can also function as a receptor and thus may be important in cell signaling. The present minireview will focus on the role of tissue factor in the pathogenesis of septic shock, infectious endocarditis and invasive aspergillosis, as determined by both in vivo and in vitro models. PMID- 12886453 TI - Sulfated proteoglycans as modulators of neuronal migration and axonal decussation in the developing midbrain. AB - Proteoglycans are abundant in the developing brain and there is much circumstantial evidence for their roles in directional neuronal movements such as cell body migration and axonal growth. We have developed an in vitro model of astrocyte cultures of the lateral and medial sectors of the embryonic mouse midbrain, that differ in their ability to support neuritic growth of young midbrain neurons, and we have searched for the role of interactive proteins and proteoglycans in this model. Neurite production in co-cultures reveals that, irrespective of the previous location of neurons in the midbrain, medial astrocytes exert an inhibitory or nonpermissive effect on neuritic growth that is correlated to a higher content of both heparan and chondroitin sulfates (HS and CS). Treatment of astrocytes with chondroitinase ABC revealed a growth-promoting effect of CS on lateral glia but treatment with exogenous CS-4 indicated a U shaped dose-response curve for CS. In contrast, the growth-inhibitory action of medial astrocytes was reversed by exogenous CS-4. Treatment of astrocytes with heparitinase indicated that the growth-inhibitory action of medial astrocytes may depend heavily on HS by an as yet unknown mechanism. The results are discussed in terms of available knowledge on the binding of HS proteoglycans to interactive proteins, with emphasis on the importance of unraveling the physiological functions of glial glycoconjugates for a better understanding of neuron-glial interactions. PMID- 12886454 TI - Functional role of a specific ganglioside in neuronal migration and neurite outgrowth. AB - Cell migration occurs extensively during mammalian brain development and persists in a few regions in the adult brain. Defective migratory behavior of neurons is thought to be the underlying cause of several congenital disorders. Knowledge of the dynamics and molecular mechanisms of neuronal movement could expand our understanding of the normal development of the nervous system as well as help decipher the pathogenesis of neurological developmental disorders. In our studies we have identified and characterized a specific ganglioside (9-O-acetyl GD3) localized to the membrane of neurons and glial cells that is expressed in regions of cell migration and neurite outgrowth in the developing and adult rat nervous system. In the present article we review our findings that demonstrate the functional role of this molecule in neuronal motility. PMID- 12886455 TI - Thymocyte migration: an affair of multiple cellular interactions? AB - Cell migration is a crucial event in the general process of thymocyte differentiation. The cellular interactions involved in the control of this migration are beginning to be defined. At least chemokines and extracellular matrix proteins appear to be part of the game. Cells of the thymic microenvironment produce these two groups of molecules, whereas developing thymocytes express the corresponding receptors. Moreover, although chemokines and extracellular matrix can drive thymocyte migration per se, a combined role for these molecules appears to contribute to the resulting migration patterns of thymocytes in their various stages of differentiation. The dynamics of chemokine and extracellular matrix production and degradation is not yet well understood. However, matrix metalloproteinases are likely to play a role in the breakdown of intrathymic extracellular matrix contents. Thus, the physiological migration of thymocytes should be envisioned as a resulting vector of multiple, simultaneous and/or sequential stimuli involving chemokines, adhesive and de-adhesive extracellular matrix proteins, as well as matrix metalloproteinases. Accordingly, it is conceivable that any pathological change in any of these loops may result in the alteration of normal thymocyte migration. This seems to be the case in murine infection by the protozoan parasite Trypanosoma cruzi, the causative agent of Chagas' disease. A better knowledge of the physiological mechanisms governing thymocyte migration will provide new clues for designing therapeutic strategies targeting developing T cells. PMID- 12886456 TI - Building bone tissue: matrices and scaffolds in physiology and biotechnology. AB - Deposition of bone in physiology involves timed secretion, deposition and removal of a complex array of extracellular matrix proteins which appear in a defined temporal and spatial sequence. Mineralization itself plays a role in dictating and spatially orienting the deposition of matrix. Many aspects of the physiological process are recapitulated in systems of autologous or xenogeneic transplantation of osteogenic precursor cells developed for tissue engineering or modeling. For example, deposition of bone sialoprotein, a member of the small integrin-binding ligand, N-linked glycoprotein family, represents the first step of bone formation in ectopic transplantation systems in vivo. The use of mineralized scaffolds for guiding bone tissue engineering has revealed unexpected manners in which the scaffold and cells interact with each other, so that a complex interplay of integration and disintegration of the scaffold ultimately results in efficient and desirable, although unpredictable, effects. Likewise, the manner in which biomaterial scaffolds are "resorbed" by osteoclasts in vitro and in vivo highlights more complex scenarios than predicted from knowledge of physiological bone resorption per se. Investigation of novel biomaterials for bone engineering represents an essential area for the design of tissue engineering strategies. PMID- 12886457 TI - Collagens and proteoglycans of the corneal extracellular matrix. AB - The cornea is a curved and transparent structure that provides the initial focusing of a light image into the eye. It consists of a central stroma that constitutes 90% of the corneal depth, covered anteriorly with epithelium and posteriorly with endothelium. Its transparency is the result of the regular spacing of collagen fibers with remarkably uniform diameter and interfibrillar space. Corneal collagen is composed of heterotypic fibrils consisting of type I and type V collagen molecules. The cornea also contains unusually high amounts of type VI collagen, which form microfibrillar structures, FACIT collagens (XII and XIV), and other nonfibrillar collagens (XIII and XVIII). FACIT collagens and other molecules, such as leucine-rich repeat proteoglycans, play important roles in modifying the structure and function of collagen fibrils.Proteoglycans are macromolecules composed of a protein core with covalently linked glycosaminoglycan side chains. Four leucine-rich repeat proteoglycans are present in the extracellular matrix of corneal stroma: decorin, lumican, mimecan and keratocan. The first is a dermatan sulfate proteoglycan, and the other three are keratan sulfate proteoglycans. Experimental evidence indicates that the keratan sulfate proteoglycans are involved in the regulation of collagen fibril diameter, and dermatan sulfate proteoglycan participates in the control of interfibrillar spacing and in the lamellar adhesion properties of corneal collagens. Heparan sulfate proteoglycans are minor components of the cornea, and are synthesized mainly by epithelial cells. The effect of injuries on proteoglycan synthesis is discussed. PMID- 12886458 TI - Antioxidant dietary deficiency induces caspase activation in chick skeletal muscle cells. AB - Apoptosis and necrosis are two distinct forms of cell death that can occur in response to different agents and stress conditions. In order to verify if the oxidative stress induced by dietary selenium and vitamin E deficiencies can lead muscle cells to apoptosis, one-day-old chicks were reared using diets differing in their vitamin E (0 or 10 IU/kg) and selenium (0 or 0.15 ppm) supplementation. Chick skeletal muscle tissue was obtained from 28-day-old animals and used to verify apoptosis occurrence based on caspase activity detection and DNA fragmentation. Antioxidant deficiency significantly increased caspase-like activity assessed by the hydrolysis of fluorogenic peptide substrates (Abz peptidyl-EDDnp) at lambda exc = 320 nm and lambda em = 420 nm. Proteolytic activation was not accompanied by typical internucleosomal DNA fragmentation detected by field inversion gel electrophoresis. Although the general caspase inhibitor N-benzyloxycarbonyl-Val-Ala-Asp(O-Me) fluoromethyl ketone (Z-VAD-fmk) (0 to 80 muM) did not block caspase-like activity when preincubated for 30 min with muscle homogenates, the hydrolyzed substrates presented the same cleavage profile in HPLC (at the aspartic acid residue) when incubated with the purified recombinant enzyme caspase-3. These data indicate that oxidative stress causes caspase-like activation in muscle cells and suggest that cell death associated with exudative diathesis (dietary deficiency of selenium and vitamin E) can follow the apoptotic pathway. PMID- 12886459 TI - Glycosaminoglycans affect the interaction of human plasma kallikrein with plasminogen, factor XII and inhibitors. AB - Human plasma kallikrein, a serine proteinase, plays a key role in intrinsic blood clotting, in the kallikrein-kinin system, and in fibrinolysis. The proteolytic enzymes involved in these processes are usually controlled by specific inhibitors and may be influenced by several factors including glycosaminoglycans, as recently demonstrated by our group. The aim of the present study was to investigate the effect of glycosaminoglycans (30 to 250 micro/ml) on kallikrein activity on plasminogen and factor XII and on the inhibition of kallikrein by the plasma proteins C1-inhibitor and antithrombin. Almost all available glycosaminoglycans (heparin, heparan sulfate, bovine and tuna dermatan sulfate, chondroitin 4- and 6-sulfates) reduced (1.2 to 3.0 times) the catalytic efficiency of kallikrein (in a nanomolar range) on the hydrolysis of plasminogen (0.3 to 1.8 microM) and increased (1.9 to 7.7 times) the enzyme efficiency in factor XII (0.1 to 10 microM) activation. On the other hand, heparin, heparan sulfate, and bovine and tuna dermatan sulfate improved (1.2 to 3.4 times) kallikrein inhibition by antithrombin (1.4 microM), while chondroitin 4- and 6 sulfates reduced it (1.3 times). Heparin and heparan sulfate increased (1.4 times) the enzyme inhibition by the C1-inhibitor (150 nM). PMID- 12886460 TI - Distribution of biglycan and decorin in rat dental tissue. AB - Biglycan and decorin are small leucine-rich proteoglycans that play several biological and structural roles in different tissues and organs. Several reports have indicated that biglycan participates in odontoblast and ameloblast differentiation and in the calcification process. In the present study we show that the expression of biglycan changes from within the ameloblasts and odontoblasts to the extracellular space according to the stage of animal development. In predentin and in the pulp space, however, biglycan was continually expressed throughout the period of investigation. In contrast, decorin was absent in odontoblasts and in ameloblasts and was exclusively expressed in predentin throughout the period of observation. In young rats, however, decorin was expressed in the extracellular spaces of the pulp, where it was concentrated mainly in the peripheral pulp. PMID- 12886461 TI - Distribution of versican and hyaluronan in the mouse uterus during decidualization. AB - Preparation for embryo implantation requires extensive adaptation of the uterine microenvironment. This process consists of cell proliferation and cell differentiation resulting in the transformation of endometrial fibroblasts into a new type of cell called decidual cell. In the present study, we followed the space-time distribution of versican and hyaluronan (HA) in different tissues of the uterus before and after embryo implantation. Fragments of mouse uteri obtained on the fourth, fifth, sixth and seventh days of pregnancy were fixed in Methacarn, embedded in Paraplast and cut into 5-microm thick sections. HA was detected using a biotinylated fragment of the proteoglycan aggrecan, which binds to this glycosaminoglycan with high affinity and specificity. Versican was detected by a polyclonal antibody. Both reactions were developed by peroxidase methods. Before embryo implantation, both HA and versican were present in the endometrial stroma. However, after embryo implantation, HA disappeared from the decidual region immediately surrounding the implantation chamber, whereas versican accumulated in the same region. The differences observed in the expression of HA and versican suggest that both molecules may participate in the process of endometrial decidualization and/or embryo implantation. PMID- 12886462 TI - Immunohistochemical demonstration of TGF-beta and decorin in paracoccidioidal granulomas. AB - Different patterns of granulomas have been observed in 6- to 8-week-old mice after ip inoculation with 5 x 10(6) yeast cells of Paracoccidioides brasiliensis. Transforming growth factor-beta (TGF-beta) is a cytokine that has been shown to participate in fibrosis and granuloma formation; its activities seem to be modulated by the small proteoglycan decorin. In the present study, TGF-beta and decorin expression in epiploon granulomas was assessed by immunohistochemistry in susceptible (B10.A) and resistant (A/J) mice after 15, 30, 120 and 150 days of P. brasiliensis ip infection. The epiploon was collected, fixed in Methacarn solution and embedded in paraffin, and 5-microm thick sections were used for immunohistochemical analysis employing the streptavidin-biotin-peroxidase technique. The former mouse strain developed fatal disease with many disseminated lesions increasing in size and number during the infection and the latter developed mild disease with the presence of encapsulated granulomas. In the epiploon, TGF-beta was present on macrophages, giant cells, lymphocytes and fibroblasts, and absent on neutrophils. It was also detected in areas of fibrosis and necrosis, as well as disperse in amorphous extracellular matrix, mostly in resistant mice. Decorin was present circumscribing macrophages and giant cells containing fungi, but absent on these cells. In both mouse strains, decorin was found at the periphery of the lesions, and markedly in milky spot granulomas. In resistant mice, positivity was found around fibrotic and necrotic areas of encapsulated and residual lesions containing lysed fungi. Decorin was found associated with thick fibers around encapsulated lesions. In susceptible mice, the size and number of lesions increased with the progression of the disease and were correlated with the weaker expression of decorin. We suggest an association of decorin with the fibrogenic process observed in paracoccidioidal granulomas. PMID- 12886463 TI - Decorin is one of the proteoglycans expressed in Walker 256 rat mammary carcinoma. AB - Proteoglycan and glycosaminoglycan content was analyzed in a model of rat mammary carcinoma to study the roles of these compounds in tumorigenesis. Hyaluronic acid and proteoglycans bearing chondroitin and/or dermatan sulfate chains were detected in solid tumors obtained after subcutaneous inoculation of Walker 256 rat carcinoma cells. About 10% of sulfated glycosaminoglycan chains corresponded to heparan sulfate. The small leucine-rich proteoglycan, decorin, was identified as one of the proteoglycans, in addition to others of higher molecular weight, by cross-reaction with an antiserum raised against pig laryngeal decorin and by N terminal amino acid sequencing. Decorin was separated from other proteoglycans by hydrophobic chromatography and its complete structure was determined. It has a molecular weight of about 85 kDa and a dermatan chain of 45 kDa with 4-sulfated disaccharides. After degradation of the glycosaminoglycan chain, three core proteins of different molecular weight (36, 46 and 56 kDa) were identified. The presence of hyaluronic acid and decorin has been reported in a variety of tumors and tumor cells. In the Walker 256 mammary carcinoma model, hyaluronic acid may play an important role in tumor progression, since it provides a more hydrated extracellular matrix. On the other hand, decorin, which is expressed by stromal cells, represents a host defense response to tumor growth. PMID- 12886464 TI - Simultaneous changes in the function and expression of beta 1 integrins during the growth arrest of poorly differentiated colorectal cells (LISP-1). AB - Cells usually lose adhesion and increase proliferation and migration during malignant transformation. Here, we studied how proliferation can affect the other two characteristics, which ultimately lead to invasion and metastasis. We determined the expression of beta 1 integrins, as well as adhesion and migration towards laminin-1, fibronectin, collagens type I and type IV presented by LISP-1 colorectal cancer cells exposed to 2.5% dimethyl sulfoxide (DMSO), an agent capable of decreasing proliferation in this poorly differentiated colorectal cell line. Untreated cells (control), as shown by flow cytometry and monoclonal antibodies, expressed alpha 2 (63.8 11.3% positive cells), alpha 3 (93.3 7.0%), alpha 5 (50.4 12.0%) and alpha 6 (34.1 4.9%) integrins but not alpha1, alpha 4, alpha v or 4. Cells adhered well to laminin-1 (73.4 6.0%) and fibronectin (40.0 2.0%) substrates but very little to collagens. By using blocking monoclonal antibodies, we showed that alpha 2, alpha 3 and alpha 6 mediated laminin-1 adhesion, but neither alpha 3 nor alpha 5 contributed to fibronectin adherence. DMSO arrested cells at G0/G1 (control: 55.0 2.4% vs DMSO: 70.7 2.5%) while simultaneously reducing alpha 5 (24.2 19%) and alpha 6 (14.3 10.8%) expression as well as c-myc mRNA (7-fold), the latter shown by Northern blotting. Although the adhesion rate did not change after exposure to DMSO, alpha 3 and alpha 5 played a major role in laminin-1 and fibronectin adhesion, respectively. Migration towards laminin-1, which was clearly increased upon exposure to DMSO (control: 6 2 cells vs DMSO: 64 6 cells), was blocked by an antibody against alpha 6. We conclude that the effects of DMSO on LISP-1 proliferation were accompanied by concurrent changes in the expression and function of integrins, consequently modulating adhesion/migration, and revealing a complex interplay between function/expression and the proliferative state of cells. PMID- 12886465 TI - Differential expression of integrin subunits on adherent and nonadherent mast cells. AB - Mast cell progenitors arise in bone marrow and then migrate to peripheral tissues where they mature. It is presumed that integrin receptors are involved in their migration and homing. In the present study, the expression of various integrin subunits was investigated in three systems of adherent and nonadherent mast cells. Mesentery mast cells, freshly isolated bone marrow-derived mast cells (BMMC) and RBL-2H3 cells grown attached to tissue culture flasks are all adherent mast cells and peritoneal mast cells, and cultured BMMC and RBL-2H3 cells grown in suspension represent nonadherent mast cell populations. Pure populations of mast cells were immunomagnetically isolated from bone marrow, mesentery and peritoneal lavage using the mast cell-specific monoclonal antibody AA4. By immunomicroscopy, we could demonstrate that all of these mast cells expressed alpha 4, alpha 5, alpha 6, beta 1 and beta 7 integrin subunits. The expression of the alpha 4 integrin subunit was 25% higher in freshly isolated mesentery mast cells and BMMC. Consistent with the results obtained by immunomicroscopy, mesentery mast cells expressed 65% more mRNA for the alpha 4 integrin subunit than peritoneal mast cells. In vitro studies were also conducted using the rat mast cell line RBL-2H3. RBL-2H3 cells grown attached to the tissue culture flasks or as suspension cultures expressed the same integrin subunits identified in bone marrow, mesenteric and peritoneal mast cells ex vivo. Similarly, the expression of alpha 4 integrin was higher in adherent cells. Therefore, alpha 4 integrins may play a critical role in the anchorage of mast cells to the extracellular matrix in bone marrow and in peripheral tissues. PMID- 12886466 TI - Changes in cell shape, cytoskeletal proteins and adhesion sites of cultured cells after extracellular Ca2+ chelation. AB - Although much is known about the molecules involved in extracellular Ca2+ regulation, the relationship of the ion with overall cell morphology is not understood. The objective of the present study was to determine the effect of the Ca2+ chelator EGTA on the major cytoskeleton components, at integrin-containing adhesion sites, and their consequences on cell shape. Control mouse cell line C2C12 has a well-spread morphology with long stress fibers running in many different directions, as detected by fluorescence microscopy using rhodamine phalloidin. In contrast, cells treated with EGTA (1.75 mM in culture medium) for 24 h became bipolar and showed less stress fibers running in one major direction. The adhesion plaque protein alpha 5-integrin was detected by immunofluorescence microscopy at fibrillar adhesion sites in both control and treated cells, whereas a dense labeling was seen only inside treated cells. Microtubules shifted from a radial arrangement in control cells to a longitudinal distribution in EGTA treated cells, as analyzed by immunofluorescence microscopy. Desmin intermediate filaments were detected by immunofluorescence microscopy in a fragmented network dispersed within the entire cytoplasm in EGTA-treated cells, whereas a dense network was seen in the whole cytoplasm of control cells. The present results suggest that the role of extracellular Ca2+ in the regulation of C2C12 cell shape can be mediated by actin-containing stress fibers and microtubules and by intermediate filament reorganization, which may involve integrin adhesion sites. PMID- 12886467 TI - Cytoskeletal and cellular adhesion proteins in zebrafish (Danio rerio) myogenesis. AB - The current myogenesis and myofibrillogenesis model has been based mostly on in vitro cell culture studies, and, to a lesser extent, on in situ studies in avian and mammalian embryos. While the more isolated artificial conditions of cells in culture permitted careful structural analysis, the actual in situ cellular structures have not been described in detail because the embryos are more difficult to section and manipulate. To overcome these difficulties, we used the optically clear and easy to handle embryos of the zebrafish Danio rerio. We monitored the expression of cytoskeletal and cell-adhesion proteins (actin, myosin, desmin, alpha-actinin, troponin, titin, vimentin and vinculin) using immunofluorescence microscopy and video-enhanced, background-subtracted, differential interference contrast of 24- to 48-h zebrafish embryos. In the mature myotome, the mononucleated myoblasts displayed periodic striations for all sarcomeric proteins tested. The changes in desmin distribution from aggregates to perinuclear and striated forms, although following the same sequence, occurred much faster than in other models. All desmin-positive cells were also positive for myofibrillar proteins and striated, in contrast to that which occurs in cell cultures. Vimentin appeared to be striated in mature cells, while it is developmentally down-regulated in vitro. The whole connective tissue septum between the somites was positive for adhesion proteins such as vinculin, instead of the isolated adhesion plaques observed in cell cultures. The differences in the myogenesis of zebrafish in situ and in cell culture in vitro suggest that some of the previously observed structures and protein distributions in cultures could be methodological artifacts. PMID- 12886468 TI - Trypanosoma cruzi infection: a continuous invader-host cell cross talk with participation of extracellular matrix and adhesion and chemoattractant molecules. AB - Several lines of evidence have shown that Trypanosoma cruzi interacts with host extracellular matrix (ECM) components producing breakdown products that play an important role in parasite mobilization and infectivity. Parasite-released antigens also modulate ECM expression that could participate in cell-cell and/or cell-parasite interactions. Increased expression of ECM components has been described in the cardiac tissue of chronic chagasic patients and diverse target tissues including heart, thymus, central nervous system and skeletal muscle of experimentally T. cruzi-infected mice. ECM components may adsorb parasite antigens and cytokines that could contribute to the establishment and perpetuation of inflammation. Furthermore, T. cruzi-infected mammalian cells produce cytokines and chemokines that not only participate in the control of parasitism but also contribute to the establishment of chronic inflammatory lesions in several target tissues and most frequently lead to severe myocarditis. T. cruzi-driven cytokines and chemokines may also modulate VCAM-1 and ICAM-1 adhesion molecules on endothelial cells of target tissues and play a key role in cell recruitment, especially of activated VLA-4+LFA-1+CD8+ T lymphocytes, resulting in a predominance of this cell population in the inflamed heart, central nervous system and skeletal muscle. The VLA-4+-invading cells are surrounded by a fine network of fibronectin that could contribute to cell anchorage, activation and effector functions. Since persistent "danger signals" triggered by the parasite and its antigens are required for the establishment of inflammation and ECM alterations, therapeutic interventions that control parasitism and selectively modulate cell migration improve ECM abnormalities, paving the way for the development of new therapeutic strategies improving the prognosis of T. cruzi-infected individuals. PMID- 12886469 TI - [In support of an antibarbarus, a scholarly apparatus of bad radiologic idioms]. PMID- 12886470 TI - [Embolization of symptomatic myomas (UAE): technique, indication and results]. AB - Presentation of indication, technique and results of transarterial uterine artery embolization (UAE) for the treatment of symptomatic myomas. Technical requirements are presented like DSA, catheters, superselective catheterisation and the different embolization materials as polyvinylalcohol (PVA) or microspheres, as well as the follow-up after UAE. The technical success rate of UAE is documented to range between 98 to 100 % and myomatous symptomatology disappears in 85 to 94 % of the cases. A reduction in the size of the myomata after UAE is observed between 48 to 70 %. A resolution of the hemorrhage disappears in 80 to 96 % of the cases immediately. Particulate embolization of the uterine artery is a new minimally invasive therapy in the management of symptomatic leiomyomas with a high efficiency and low rate of major complications. Further studies may prove the longterm results after UAE, the influence on fertility and possible late complications. PMID- 12886471 TI - [Preoperative functional magnetic resonance tomography (FMRI) in patients with rolandic brain tumors: indication, investigation strategy, possibilities and limitations of clinical application]. AB - Preoperative functional magnetic resonance imaging (fMRI) localizes the primary motor and somatosensory cortex in relation to rolandic brain tumors and determines plastic cortical reorganization. Functional landmarks help to assess the indication for surgery and to plan for safer surgical procedures that protect the functional cortex during resection even when morphologic landmarks are no longer identifiable on anatomic images. Despite its successful application, preoperative fMRI has not yet reached the status of an established clinical diagnostic procedure since special stimulation systems, standardized fMRI protocols and medically approved software are still lacking. Following a brief review of the image display of the functional and morphologic anatomy, the different indications for preoperative fMRI in patients with rolandic brain tumors are presented. A robust preoperative protocol enables clinical MR units with magnetic field strengths of 1.0 Tesla or higher to perform reliable fMRI during contralateral hand movements. Optimized investigation strategies and stimulation modalities are proposed for patients with rolandic tumors distant from the cortical hand representation, for patients with preexisting sensorimotor deficits and for patients with poor compliance. Representative cases illustrate the clinical application. Possibilities and limitations of preoperative fMRI are presented and discussed. PMID- 12886472 TI - Estimation of radiation exposure in 16-detector row computed tomography of the heart with retrospective ECG-gating. AB - PURPOSE: We sought to evaluate the radiation exposure of cardiac CT scans with 16 row multidetector computed tomography (MDCT). Additionally the possibility of dose reduction by using a ECG-controlled tube current modulation technique was evaluated. METHODS AND MATERIAL: An Alderson Rando phantom equipped with thermoluminescent dosimeters was used for dose measurements. Effective dose was calculated according to ICRP 60. Exposure was performed on a 16-row MDCT scanner with standard protocols for CT coronary calcium scoring (120 kV, 133 mAs, 12 x 1.5 mm) and CT coronary angiography (120 kV, 400 mAs, 12 x 0.75 mm). Exposure was repeated at a simulated heart rate of 60 bpm with ECG-pulsed tube current modulation. RESULTS: Effective dose was 2.9 mSv (male) and 3.6 mSv (female) for the calcium scoring protocol. CT coronary angiography resulted in an effective dose of 8.1 mSv (male) and 10.9 mSv (female). Using ECG-pulsed tube current modulation radiation exposure can significant reduced: by 46 % (1.6 mSv) in calcium scoring and by 47 % (4.3 mSv) in CT coronary angiography. CONCLUSION: MDCT of the heart shows a significant radiation exposure, which can significantly be reduced by ECG-pulsed tube current modulation. Radiation exposure of cardiac MDCT is comparable to CT-examinations of chest or abdomen, but seem to be slightly higher compared to conventional coronary angiography. PMID- 12886473 TI - [Focal cortical dysplasias: neuroradiological findings and differential diagnosis]. AB - PURPOSE: To describe MRI findings of four types of focal cortical dysplasia (FCD) and compare them with diagnostic criteria reported in the literature. MATERIAL AND METHODS: This study includes eight patients with seizures in whom cranial MRI diagnosed an FCD, with histologic confirmation in two patients. RESULTS: In all patients, the dysplastic cortex was thickened. Its signal was hyperintense on T 2 -weighted and FLAIR images, but variable on T 1 -weighted images. Blurring of the corticomedullary junction was present in 5 patients. In one patient, MRI demonstrated vascular proliferation within the FCD. Type l or II FCD was diagnosed in six patients, and transmantle FCD and type IV FCD with capillary proliferation in one patient each. DISCUSSION: Thickening and hyperintensity of the cortex on T 2 -weighted and FLAIR images are more reliable signs of FCD than blurring of the gray matter-white matter junction and signal changes on T 1 weighted images. Typical vascular proliferation may be detectable with MRI and suggests the diagnosis of FCD with glial proliferation. Edema, calcification, or pathologic contrast enhancement has not been observed in FCD to date. PMID- 12886474 TI - [Intracranial aneurysms: pathogenesis, rupture risk, treatment options]. AB - Non-invasive or minimal invasive imaging techniques, such as CTA and MRA, are widely used and lead to an increasing number of incidentally diagnosed intracranial aneurysms. This review answers questions regarding pathogenesis, risk of rupture and treatment options. In conclusion, treatment guidelines based on the individual's risk profile are presented. PMID- 12886475 TI - [Spinal dural arteriovenous fistula: clinical and radiological findings in 54 patients]. AB - PURPOSE: Presentation of the clinical and radiological findings in spinal dural arteriovenous fistulas (SDAVF) based on the experience in 54 patients. MATERIAL AND METHODS: Evaluation of patients' records and myelography (n = 23), MRI (n = 54) as well as conventional angiography (n = 54) with respect to history, symptoms, clinical and radiological results. RESULTS: Clinically, a long history (mean 20 months) with progressive ascending paresis (100 %), sensory deficits (93 %) and loss of control over bladder and bowel function (89 %) in male patients (78 %) of advanced age (mean 60 years) indicates the disease. Typical signs in MRI are central cord hyperintensity (100 %) with slight medullary distension (74 %), contrast enhancement (79 %) and distended perimedullary veins (89 %), the latter being disclosed by myelography in 78 % of cases. For diagnosis spinal angiography is necessary which most often shows a thoracic location of the fistula (69 %), more than one feeding artery (67 %) and caudal as well as rostral venous drainage (63 %). CONCLUSION: MRI is superior to myelography to detect diagnostic signs of SDAVF in patients with typical clinical presentation. For definitive diagnosis spinal angiography is still indispensable. PMID- 12886476 TI - [Diagnostic impact of multiplanar reformations in multi-slice CT of laryngeal and hypopharyngeal carcinomas]. AB - PURPOSE: Assessment of the diagnostic value of multiplanar reformations (MPR) in multi-slice computed tomography (MSCT) by comparing relevant anatomic structures of the larynx and hypopharynx. MATERIALS AND METHODS: MSCT of the neck was performed in 69 consecutive patients (including 30 laryngeal and 12 hypopharyngeal carcinomas). From a 4 x 1.0 mm collimation data set, 3.0-mm and 1.25-mm axial slices as well as 3-mm coronal and sagittal slices were reconstructed. Using the histological examination as gold standard, sensitivity and specificity regarding tumor infiltration for all relevant anatomical structures of the larynx and the hypopharynx were determined for each reconstruction and compared with the McNemar test. Moreover, 42 patients with laryngeal and/or hypopharyngeal carcinoma were subjectively evaluated to determine whether the respective reconstructions enables a better topographical visualization of the tumor in relation to surrounding structures and, furthermore, whether this has an influence on the therapeutical strategy (operation versus radiation therapy, type of operation, surgical approach). RESULTS: Sensitivities and specificities were not significantly different between the reconstructions. However, coronal and sagittal MPR provided a better topographical visualization of the tumor in 14 of 42 (33 %) of the patients, and influenced the therapeutical strategy in 8 of 42 (19 %) of the patients. A lowered signal-to-noise ratio impeded the evaluation of the relatively thin 1.25 mm axial slices in more than 23 % of the cases. CONCLUSION: Besides the 3-mm axial slices, coronal and sagittal MPR can improve the topographical visualization of laryngopharyngeal tumors and are recommended for preoperative MSCT of laryngeal and/or hypopharyngeal carcinomas. Additionally reconstructed 1.25-mm axial slices can be discarded since they do not hold a higher value compared to 3-mm axial reconstructions. PMID- 12886477 TI - [Evaluation of passive cardiomyoplasty using left- and right-ventricular volume measurements by EBCT and MRI in patients with chronic congestive heart failure]. AB - PURPOSE: To evaluate the efficacy of passive cardiomyoplasty with the determination of biventricular volumes, global systolic function as well as left ventricular muscle mass. MATERIALS AND METHODS: In 19 patients with congestive heart failure of idiopathic or ischemic origin, a polyester mesh-graft was implanted around both ventricles for stabilization and functional support. Before and three months after surgery, 15 patients underwent EBCT and 4 patients with impaired renal function underwent MRI, for the evaluation of the volume and ejection fraction of both ventricles. RESULTS: EBCT demonstrated a decrease from 385 to 310 ml in LV-EDV, from 312 to 242 ml in LV-ESV, from 209 to 160 ml in RV EDV and from 149 to 87 ml in RV-ESV, and an increase from 20 to 26% in LV-EF and from 37 to 50% in RV-EF as well as a reduction of LV-MM from 300 to 274 g (p < 0.05 each). Similar results were obtained by MRI. CONCLUSION: Following passive cardiomyoplasty, EBCT and MRI revealed an improvement of the global systolic function as well as a reduction of biventricular volumes and left-ventricular muscle mass. PMID- 12886478 TI - [Quantifying the inflammatory activity in Crohn's disease using CE dynamic MRI]. AB - PURPOSE: Evaluation of dynamic contrast enhanced MRI in patients with Crohn's disease to assess local inflammatory activity. MATERIAL AND METHODS: Prospective study of 13 patients with histologically proven Crohn's disease. Axial and coronal slices were acquired by a 1.5 T MR (Magnetom Vision, Siemens, Germany): T1 flash 2 D (TR 72.5 ms, TE 4.1 ms), T2 (TR 2730 ms, TE 138 ms), turbo-flash sequences T1 (TR 94.2 ms, TE 4.1 ms) post contrast media fat saturated (Magnevist, 0.2 ml/kg, flow 4 ml/s). In area of maximal thickening of terminal ileal wall, axial dynamic T1 sequences (TR 11 ms, TE 4.2 ms) were acquired every 1.5 s post contrast media application for a total duration of 1 min. Contrast uptake was subjectively measured by semiquantitative score and computed assisted ROI evaluation. MR parameters were correlated with CDAI (Crohn's disease activity index) and SAI (severe activity index). RESULTS: Contrast uptake in the intestinal wall occurred after 18.5 s (range: 3.0 - 28.0), contrast upslope until plateau phase lasted for 16.1 s (range: 8.0 - 50.0). Maximum contrast enhancement into the bowel wall was 266 % (105 - 450 %) of baseline. After maximum contrast uptake, we observed a plateau phase in all cases for the total duration of measurement. A significant correlation existed for maximum contrast uptake to CDAI (r = 0.591; p = 0.033), for beginning of contrast upslope to the time until plateau phase (r = 0.822; p = 0.001), and for the time until plateau phase to CDAI (r = 0.562; p = 0.046). CDAI was on average 108, median 106; SAI was on average 114, median 115. SAI correlated significantly to CDAI (r = 0.874). Maximum contrast uptake, beginning of contrast upslope, and time until plateau phase were independent to creeping fat, local lymphadenitis, laboratory parameters, temperature, body mass index, heart frequency and systolic blood pressure. CONCLUSION: Dynamic MRI enables to quantify local inflammatory activity of bowel wall in patients with Crohn's disease. Larger studies are necessary to establish this method in clinical routine. PMID- 12886479 TI - [Functional MRI of the pelvic floor: postpartum changes of primiparous women after spontaneous vaginal delivery]. AB - PURPOSE: Detection of morphological and functional changes of the pelvic floor with functional MRI in primiparous women after spontaneous vaginal delivery. METHODS AND MATERIALS: The study comprises 26 primiparous women after vaginal delivery and a control group of 41 healthy asymptomatic nulliparous volunteers. MRI was performed on a 1.5 T system in supine position with vagina and rectum opacified with Sonogel. The static images consisted of sagittal and axial T 2 weighted SE sequences and functional images of true FISP sequences in midsagittal and axial planes acquired with the patient at rest, straining and during defecation. Evaluation of morphometric parameters included pelvimetry, thickness of the puborectal muscle and width of the urogenital hiatus as well as position and movement of the pelvic organs relative to the pubococcygeal reference line. RESULTS: The configuration of the bony pelvis did not differ for both groups. The puborectal muscle was significantly thinner in the study group (0.8 cm vs 0.6 cm). The functional images showed no significant differences between both groups at rest but a significantly increased incidence in the descent of the bladder neck, vaginal fornix and anorectal junction in the study group during straining. In addition, the primiparous women had more prominent rectoceles (0.6 cm vs 1.5 cm). CONCLUSION: Static imaging alone fails to demonstrate relevant pelvic floor changes and a functional method is necessary to evaluate the interactions of the pelvic organs regarding organ descent. Functional MRI of the pelvic floor is an excellent method to reveal the significant changes of the pelvic floor after vaginal birth without exposing the uterus to radiation. PMID- 12886480 TI - [Virtual cystoscopy with reduced tube current: an alternative to cystoscopy?]. AB - PURPOSE: Virtual cystoscopy at different mAs settings in patients with tumors of the bladder in comparison with cystoscopy and axial CT. MATERIALS AND METHODS: The study included 28 patients with bladder tumors and 28 patients without tumors as control group. The bladder was distended with air and scanned at two different mAs settings (protocol 1 and 2). The radiation exposure was reduced by 82 % for protocol 2 in comparison with protocol 1. Three readers compared virtual cystoscopy with cystoscopy and axial CT as to tumor location and size. Sensitivity, specificity and accuracy were calculated. RESULTS: Of the 47 tumors seen at cystoscopy, all readers detected 45 tumors by virtual cystoscopy using protocol 1 and 44 tumors using protocol 2, with p > 0.05 in comparison with cystoscopy. Virtual cystoscopy showed higher sensitivity than axial CT (97.2 % vs. 86.5 % for protocol 1) and (96.5 % vs. 86.5 % for protocol 2) with the same specificity (100 %). The accuracy was 0.982 for protocol 1 and 0.978 for protocol 2. In comparison with virtual cystoscopy, axial CT shoved more often over- and underestimation of the tumor size. CONCLUSION: Virtual cystoscopy at reduced mAs setting is a modality suitable for the detection of polypoid tumors, but does not provide data of the mucosa and thus cannot replace cystoscopy yet. PMID- 12886481 TI - [Transjugular liver core biopsy: indications, results, and complications]. AB - PURPOSE: To evaluate benefit, feasibility, and frequency of complications with transjugular liver biopsy using a semi-automatic Tru-cut system. MATERIALS AND METHODS: Eighty-five consecutive patients (57 males, 28 females) with various liver disorders (cirrhosis [30], hepatitis [12], acute hepatopathy [34], orthotopic liver transplantation [8], hepatocellular carcinoma [1]), coagulopathies (n=71) and/or ascites (n = 46) were referred to our department for a transjugular liver biopsy. Mean age was 48 +/- 16 years (range 17 to 75 years). Success and complications were retrospectively evaluated from the radiology reports, pathology reports, and patient files. Success was defined as procuring a tissue specimen that enabled a definite histological diagnosis. The complications included thrombosis at the puncture site, hematoma, cardiac arrhythmia, capsular perforation, hemorrhage, and cardiac damage. Mortality included all deaths within 30 days after the procedure. Procedure-related mortality included all deaths related to the procedure. RESULTS: The procedure was technically successful in 80 patients (94 %) and unsuccessful in 5 patients (6 %) due to a failed hepatic vein cannulation (1 patient with Budd Chiari syndrome and total liver vein occlusion, 4 patients with unsuitable anatomy). One biopsy pass was made in 22 patients, and two passes were made in 45 and three or more passes in 14 patients, all in a single session. The sample quality was judged by the pathologist as good in 71 of 80 patients (89 %) and poor in 8 patients (10 %). A diagnosis was not possible in 1 patient. Eight procedure-related complications occurred, which were classified according to the criteria of the Society of Interventional Radiology (SIR) as minor in 5 (3 type A, 2 type B) and major in 3 (1 pneumothorax, type C, 1 nonfatal bleeding, type D, and 1 fatal bleeding, type F). Procedure-related mortality was 1 %, overall mortality 15 % (mostly due to progressive liver failure). CONCLUSION: In patients with coagulopathies, transjugular liver biopsy is a viable alternative for hepatic tissue evaluation with a good sample quality. In a small number of patients severe complications may occur. PMID- 12886482 TI - [Penile angiography and superselective embolization therapy in arterial priapism]. AB - PURPOSE: To report feasibility, benefit and complications of penile angiography and superselective penile embolization in arterial priapism. MATERIALS AND METHODS: Four consecutive patients (aged 28, 29, 40 and 49 years), who underwent penile angiography for arterial priapism (high-flow priapism) within a four-year period, were identified by a keyword search of our radiology information system. One patient had sustained a direct penile trauma (severe blow to the erected penis) and three patients suffered from recurrent spontaneous priapism. All patients had previously undergone corporeal aspiration and noradrenaline injection to achieve detumescence. Two patients had one or several unsuccessful spongiocavernous shunt procedures. RESULTS: In three of the four patients, superselective pudendal and penile angiography demonstrated pathologic arteriocavernous shunting. In two of the three patients, superselective embolization using a coaxial micro-catheter was attempted. In one of the two patients, the cavernous artery became spastic before embolization material was actually injected, inducing immediate and lasting detumescence. In the other patient, unilateral Gelfoam embolization led to immediate detumescence but the priapism recurred 12 hours after the procedure. A subsequent contralateral Gelfoam embolization was successful. Erectile function was preserved in all cases. No procedure-related complications occurred. CONCLUSION: Our experience supports the prevailing opinion found in the current literature that superselective coaxial embolization constitutes the treatment of choice in patients with high-flow priapism. Prognosis is good with high probability of preserving the erectile function. PMID- 12886483 TI - [Treatment of malignant biliary occlusion by means of transhepatic percutaneous biliary drainage with insertion of metal stents--results of an 8-year follow-up and analysis of the prognostic parameters]. AB - PURPOSE: To analyze outcome and predictive factors for patient survival and patency rates of unresectable malignant biliary obstruction treated with percutaneous transhepatic insertion of metal stents. MATERIALS AND METHODS: This is a retroslective analysis of 130 patients treated in one interventional radiological center with data collected from patient records and by telephone interviews. The procedure-related data had been prospectively documented in a computer data base. The Kaplan-Meier analysis was performed for univariate and multivariate comparison of survival and patency rates with the log-rank test used for different tumor types. Predictive factors for survival and 30-day mortality were analyzed by a stepwise logistic regression. RESULTS: Underlying causes of malignant biliary obstructions were cholangiocarcinoma in 50, pancreatic carcinoma in 29, liver metastases in 27, gallbladder carcinoma in 20, and other tumors in 4 patients. The technical success rate was 99 %, the complication rate 27 % and the 30-day mortality 11 %. Primary patency rates (406 days with a median of 207 days) did not differ significantly for different tumor types. The survival rates were significantly (p = 0.03 by log-rank test) better for patients with cholangiocarcinoma than for patients with pancreatic carcinoma and liver metastases. Multiple regression analysis revealed no predictive factor for patient survival and 30-day mortality. CONCLUSION: Percutaneous transhepatic insertion of metal biliary endoprostheses offers a good initial and long-term relief of jaundice caused by malignant biliary obstruction. Although survival rates for patients with cholangiocarcinoma are better than for other causes of malignant biliary obstruction, a clear predictive factor is lacking for patients undergoing palliative biliary stent insertion. PMID- 12886484 TI - [Is the attenuation of thromboembolic vascular occlusions a criterion for the prognosis of thrombolysis?]. AB - PURPOSE: To test whether the prognosis of systemic thrombolysis can be estimated with the attenuation values of vascular occlusions as measured with multi-slice computed tomography (MSCT). METHODS: Prior to thrombolysis with rt-PA, the attenuation values of vascular occlusions were determined with a helical MSCT using a collimation of 0.5 mm. We examined 5 patients that were 63 - 79 years old. With the help of reference values that were obtained from a phantom study we categorised the vascular occlusions according to their attenuation values as being mixed thrombi with low or high proportions of erythrocytes, or red thrombi. TOF angiographies were done before and after the thrombolytic therapy. RESULTS: The systemic therapy with rt-PA was effective in vascular occlusions that had attenuation values of red thrombi or of mixed thrombi with a high proportion of erythrocytes. The therapy showed no effect when the attenuation values matched with mixed thrombi with a low proportion of erythrocytes. Due to artefacts that were caused by beam hardening, attenuation measurements could not be performed in the posterior fossa. CONCLUSION: The presented cases are in accordance with the results of experimental studies which indicate that the efficacy of thrombolysis increases with the attenuation values of thromboembolic vascular occlusions. The predictive value of attenuation measurements by MSCT for the prognosis of systemic thrombolysis cannot be given yet. PMID- 12886485 TI - [Pleural metastases of malignant thymoma: differential pleural mesothelioma diagnosis]. PMID- 12886486 TI - [Ectopic thyroid tissue in the right mandibular angle: differential diagnosis of uncertain soft tissue swelling of the neck]. PMID- 12886487 TI - [Meningoencephalitis caused by Listeria monocytogenes]. PMID- 12886488 TI - [Brown bowel syndrome--an unusual etiology of pseudo-obstruction of the small intestine]. PMID- 12886489 TI - [e-Learning for psychosocial medicine]. PMID- 12886490 TI - [Effectiveness of out-patient systemic therapy--a multicenter study in the German speaking part of Switzerland]. AB - The present paper describes an effectiveness-study to analyze the effectiveness of systemic treatments. In the documentation phase of two years 28 therapists documented 89 systemic therapies that have been conducted in a multi person setting. The patients represent a general population of health care takers. With an average duration of 10 appointments over 40 weeks these treatments can be judged as brief psychotherapy. Good results have been achieved within this short period of treatment. The reduction of symptoms reached middle to high effect sizes in all dimensions of the SCL-90-R. Regarding the global severity index (GSI of SCL-90-R) 22.5 % of the patients can be considered as cured and 55 % as cured or improved. PMID- 12886491 TI - [Attitudes towards professional euthanasia in the range between grement in the society and personal preferences--results of a representative examination of the German general population]. AB - Several surveys of the German population concerning the attitude towards euthanasia in patients with terminal illness yielded contradictory results, ranging from high acquisition to high refusal rates. After a critical discussion of the methodological concepts of these investigations, we present the results of a representative study of 1957 German persons (age range: 14 - 96 years) which was performed by the institute USUMA in February 2001. Four different types of euthanasia were included in the study: active, passive, and indirect euthanasia as well as physician's assisted suicide. The affirmative response categories were "declared will and unbearable pain", "declared will", "referred will", and "in the responsibility of the physician". Additionally, we asked to state the personal preference in the case of an own incurable illness. The resulting frequency distributions stress the autonomy of the patients (declared will) and the legal forms of professional euthanasia (passive and indirect euthanasia), independent from the degree of pain. The rank order of the hypothetic personal preferences was: passive euthanasia (26.1 %), active euthanasia (21.1 %), indirect euthanasia (13.1 %) and assisted suicide (6.2 %). For each category the hypothetic personal will to utilize euthanasia personally is markedly lower than the consent to legalize euthanasia in the society. This points to a diminished readiness of the population to seek for euthanasia. Persons aged 60 years and above deny all types of euthanasia significantly more often than younger persons. Persons with subjectively bad health status prefer the category "in the responsibility of the physician" more often than healthy subjects. The representative study proves that there is no polarized public opinion concerning euthanasia, rather there is a picture of high complexity. PMID- 12886492 TI - [Psychosomatic stress factors in compensated and decompensated tinnitus]. AB - AIMS: In modern medical practice, chronic decompensated tinnitus is defined as a complex psychosomatic process in which mental and social factors are considered to have a determining effect on the patient's subjective response to the impairment of otological or other somatic functions attributed to tinnitus. What is still largely unknown is the interaction of the individual factors and their impact on the patient's ability to cope with tinnitus. The impact of psycho social and somatic factors on the subjective experience of patients with compensated and decompensated tinnitus is evaluated. PATIENTS AND METHODS: 53 patients with chronic tinnitus were divided into two groups, compensated and decompensated, on the basis of their subjective experience of the disorder, established according to the tinnitus questionnaire published by Goebel and Hiller. Self-assessment instruments and a survey of symptoms of somatic stress disorders were used to compare the two groups in terms of differences in the patients' mental and psycho-social behaviour, in their strategies for coping with tinnitus and in the incidence of co-morbidity. RESULTS: The patients with decompensated tinnitus suffered from more pronounced mental and social disabilities, were more prone to depression and used less effective techniques to cope with their illness. The principal difference between the two groups, however, appeared to lie in a significantly higher degree of somatic multi morbidity, where a particularly strong correlation was found between tinnitus and the incidence of cardiovascular diseases and hypacusis. 81 percent of the total sample of patients suffered from impaired hearing. Patients with decompensated tinnitus experienced greater communication difficulties as a result of their auditory impairment. CONCLUSIONS: In the diagnosis and therapy of tinnitus, in addition to psychic and psycho-social aspects greater attention ought to be paid to somatic factors, influencing the patient's ability to cope with the disorder. PMID- 12886493 TI - [The psychometric quality of the Nottingham Health Profile (NHP) in the general population]. AB - The Nottingham Health Profile (NHP) is a questionnaire that assesses health related quality of life. The aim of this paper is to test the psychometric quality of the instrument based on a subject sample which is representative for the German population. The sample consisted of 1996 persons with ages between 18 and 92 years. They were asked to fill in several questionnaires including the NHP. The results show that the NHP does not differentiate between subjects in the range of low and medium levels of complaints. Only one half of the subject sample agreed to at least one of the 38 items. All six subscales showed significant age and sex effects which were proved with nonlinear regression analyses. The validity was assessed through correlations with other health related questionnaires. It can be concluded that the use of the NHP cannot be recommended when studies in the general population are conducted. PMID- 12886494 TI - [Universal newborn hearing screening in Marien Hospital Hamburg from September 1999 till April 2002]. AB - BACKGROUND: With a prevalence of 5 in 1000 newborns with a hearing disorder, the congenital hearing disorders apparently constitute a serious health problem. The aim of this study was the introduction of a universal newborn hearing screening at our clinic. We also investigated if a universal screening only on parents' demand is possible. METHODS: From September 1999 to April 2002 a total of 3049 newborns, delivered at the Marien Hospital in Hamburg 97 %, were screened with the Echo-Screen TE (Fischer-Zoth). RESULTS: Only 2 of the 3049 screened infants showed signs of severe sensori-neural hearing loss. Both children belonged to high risk groups. During the period of the parental request for an investigation only 10.4 % of the newborn were screened compared to 97 % newborns during the universal newborn hearing screening. CONCLUSION: By measurement of TEOAE alone, we found a significantly lower incidence of hearing disorders than reported in literature; it may therefore not be sufficient for screening. Universal newborn hearing screening only performed on parents' demand seems to be impossible to conduct. Also before establishing a universal newborn hearing screening, cost allocation has to be solved. PMID- 12886495 TI - [Are titanium implants superior to gold implants in the tympanic cavity?]. AB - BACKGROUND: Gold implants in reconstructive surgery of the middle ear have proved to work sufficiently: That applies to tympanic cavity conditions without signs of chronic inflammation as also to the bacterially infected middle ear. For some years titanium implants are also in the use and established as well. The biocompatibility of titanium, the clinical and the audiological results are satisfactory. When using gold-prostheses in stapes surgery however, the still unsolved problem of occasional deafness has to be considered. PATIENTS: The clinical and functional results of middle ear surgery were examined retrospectively. There was no preselection of patients. 53 patients underwent middle ear surgery with titanium prostheses, 42 patients with gold prostheses. RESULTS: Due to their material properties and due to their delicate shape titanium prostheses (PORP, TORP) can be inserted into the tympanic cavity more simply and with better fit. The audiological results of gold and titanium prostheses are comparable. The extruding rate of the gold prostheses is substantial, whereas the extruding rate of titanium is negligible. CONCLUSION: The functional results of both materials are comparable. Due to the high extruding rates of gold prostheses titanium is clearly superior. PMID- 12886496 TI - [Long term results after cochlear implantation in elderly patients]. AB - BACKGROUND: Cochlear implantation has become the method of choice for the treatment of deaf patients. A better technology of stimulation opened a wider field of patients suitable for cochlear implantation. Aim of the present study was to evaluate to which extent senior patients benefit by cochlear implantation. PATIENTS AND METHODS: 36 patients between 65 and 85 years were tested by the Freiburger speech test for monosyllabic words and by the HSM sentence test with and without noise. The results were compared to those of younger implanted patients (16 - 64 years, n = 101). RESULTS: The speech perception for monosyllabic words did not differ significantly between both groups. A maximum plateau in speech perception for monosyllabic words was reached after 2 years at 55 %. The HSM sentence test revealed significant differences in the increase of speech perception between senior and younger patients in the first post surgical year. After one year elderly patients reached the same perception score as younger patients with a maximum of approximately 80 % after 3 years. At a signal to noise ratio of 15 % younger patients achieved a maximal perception of 55 % after 1 year whereas senior patients needed 4 years to reach this level. CONCLUSIONS: Senior patients benefited by cochlear implantation to the same extent as younger patients did. Senior patients gained a similar speech perception level as younger implanted patients although the elderly needed more time to reach the same scores. Nevertheless, cochlear implantation should not be denied due to age alone. PMID- 12886497 TI - [Taste disorders and recovery of the taste function after middle ear surgery]. AB - BACKGROUND: The incidence and the scale of recovery of the taste function after middle ear surgery is usually determined by the use of anamnestic data and electrogustometry. METHODS: Taste tests (regional chemical taste test, electrogustometry and subjective evaluation) were performed preoperatively, 2 to 3 weeks and 6 to 9 months after middle ear surgery. These results were evaluated by taking the scale of chorda tympani nerve (CTN) manipulation into consideration. Three groups were chosen: group 1: no or small CTN manipulation, n = 54; group 2: strong CTN manipulation, n = 14; group 3: severed CTN, n = 50. PATIENTS: In a prospective study 118 patients were examined after middle ear surgery in the ENT-Department of the University of Rostock between 2001 and 2002. 61 females and 57 males, age from 7 - 81 years, were included. RESULTS: Complaints were observed in all three groups, mainly in groups 2 and 3. These included taste disorders, taste phenomenon and numbness of the tongue. Using regional chemical taste tests, the taste loss of group 3 could be proven for those patients with complaints in 64 % and for those patients without complaints in 27 %. The electrogustometry is more sensitive than chemical taste test. All patients had pathological thresholds after chorda cutting (78 % no thresholds; 22 % elevated thresholds). 68 out of 118 patients (58 %) had a second follow-up after 6 to 9 months after middle ear surgery. The complaints are declining in all groups. In groups 2 and 3 a recovery of taste function could be observed. Even if the number of patients with severed chorda, who suffer from taste disorders, diminishes, a recovery of the taste function could only be observed in 30 %. CONCLUSIONS: The scale of chorda manipulation is important for the recovery of the taste function after middle ear surgery. The CTN should be preserved intraoperatively, especially when an operation of the contralateral ear is planned as well. PMID- 12886498 TI - [Contact endoscopic quantification of fungiform papillae--correlation to taste ability?]. AB - BACKGROUND: Besides anamnestic data and taste test, the clarification of epithelial taste disorders makes macroscopic evaluation of the surface of the tongue necessary. METHODS: In this article the contact endoscopy is evaluated for examination of the tongue epithelium and for quantification of the fungiforme papillae. These results are related to the results of the spatial taste test (1. chemical taste test with sodium chloride in 2 different suprathreshold concentrations and sucrose in also 2 different suprathreshold concentrations; 2. electrogustometry with a bipolar electrode). PATIENTS: In a prospective study we performed contact endoscopy in 16 healthy volunteers from ages 7 to 68 years. The subjects were divided into 2 age groups (< or = 45 years and > 45 years) and 2 papillae density groups. In 34 patients with either taste disorders of unknown cause or in patients with macroscopic changes at the tongue surface, the spatial and whole mouth taste tests and contact endoscopy were used to clarify the cause of the complaints. RESULTS: Density and shape of the fungiform papillae as well as the shape of the subepithelial vessels of the papillae vary among the investigated human subjects in the different age groups. No correlation between intensity ratings, papillae density und age was found after stimulation with 40 % sucrose at the tongue tip in the younger age group. Stimulation of the tongue edge with 15 % sodium chloride, however, showed a relationship between the two different density groups and the different age groups. CONCLUSIONS: The contact endoscopy allows the evaluation of the epithelium in the oral cavity. It is a nondestructive method and is additionally easy to handle, but the evaluation of the observed epithelia needs interdisciplinary co-operation with pathologists. No relevant informations are supplied in diagnostics of taste disorders. PMID- 12886499 TI - [Paranasal sinus surgery in chronic sinus disease and benign tumors indications, concepts and complications at a teaching institution]. AB - BACKGROUND AND METHODS: The medical records of 635 patients that underwent paranasal sinus surgery for chronic sinus disease or benign tumors during a 6.5 year span were assessed retrospectively to evaluate the status of sinus surgery at a teaching institution. The parameters analyzed were indication for surgery, surgical approach, extent of the procedures and complications. RESULTS: The ratio between male and female patients was 2 : 1, with an average age of 44 (+/- 16) years. In 137 cases (21.5 %), revision surgery in patients previously treated at the University of Wuerzburg was necessary. In 80 cases (12.6 %), prior surgery had been performed elsewhere. The majority of the procedures (91.3 %) were carried out using endonasal techniques. External, transethmoid (6.5 %) or transmaxillar (2.2 %) approaches were chosen in 8.7 % of all cases, mainly when the preoperative diagnoses were mucoceles (65.2 %), benign tumors (40.9 %) or orbital complications (33.3 %). Surgery was assisted exclusively by the microscope in 78.7 %, solely by the endoscope in 15.9 % and with a combination of both optical tools in 5.4 % of all cases. The total rate of complications was 8.2 %. Minor complications were seen in 8.1 % of the endonasal and 4.9 % of the external transethmoid procedures. While no major complications occurred during endonasal or extranasal transmaxillar surgery, liquorrhoe was documented in three extranasal transethmoid procedures (7.3 %). Neither permanent impairment of vision, nor post-operative meningitis nor surgery-related mortality was observed in any case. CONCLUSION: The study gives an overview over the paranasal sinus surgeries performed at a teaching institution, independent of the experience of the surgeon. It confirms the results of previous investigations and indicates that endonasal sinus surgery is a reproducible and reliable procedure that can be safely applied at a teaching institution. The results also show that the indication for extranasal approaches is further reduced to the less invasive endonasal techniques. PMID- 12886500 TI - [Preliminary results for superficial parotidectomy using the ultrasonically activated scalpel (Ultracision Harmonic Scalpel)]. AB - BACKGROUND: The high density of blood vessels in the parotid gland, the direct vicinity to the facial nerve and the narrow surgical situs require efficient intra- and postoperative hemostasis. The ultrasonic scalpel (US) accomplishes both tissue dissection and vessel coagulation simultaneously by an ultrasonically activated shear movement of 55 500 Hz. The combination of hemostasis and tissue dissection particularly qualifies US for use in surgery of the parotid gland. Until now, there have been no published studies about application of US in surgery of the parotid gland. METHOD: 20 patients with benign parotid tumors treated with a conventional technique of superficial parotidectomy and 20 patients with a superficial parotidectomy using the ultrasonically activated scalpel were examined in regard to intra- and postoperative hemostasis, wound healing and postoperative pain. Also, the quality of the histopathological specimen obtained by US was evaluated. RESULTS: In 85 % (17/20) of superficial parotidectomy efficient intraoperative hemostasis did not require electrocoagulation. In 15 % (3/20) of parotid tumors additional bipolar electrocoagulation were required because of venous bleeding from vessels exceeding 2.0 mm in diameter. Postoperative bleeding did not occur at all. Wound healing was uneventful in all cases. Surgery-related postoperative pain was not intensified. Time of operation was shortened. Histopathological evaluation, especially in the margin area, was not impaired. CONCLUSION: The US offers tissue dissection with effective intra- and postoperative hemostasis. The combination of simultaneous tissue dissection and hemostasis enables a good overall view and control of the surgical site. In addition, based on the mechanical function the US has only a minimal thermal effect on neighboring tissues and enables controlled tumor resection without damaging the facial nerve. PMID- 12886501 TI - [Tinnitus after head and neck trauma caused by traffic accident]. PMID- 12886502 TI - [Visual-vestibular interactions during linear vertical acceleration]. PMID- 12886503 TI - [Infection immunology in silicone hydrogel contact lenses for continuous wear--a review]. AB - BACKGROUND: The patient's wish for permanent refractive solution and optimal comfort must be weighed against the risk of infections. In order to meet these wishes and the demand for minimising associated risks, hyperoxygen-transmissible silicone hydrogel contact lenses have been developed. METHODS: Colonisation and sensitivity to antibiotics of bacteria in vitro, expression of bacterial ligands of the corneal epithelium (rabbit model), and the morphology of complications in patients have been reported to characterise HOTL. RESULTS: Bacterial adherence to hydrophobic silicone elastomers is low. The reduction of sensitivity towards antibiotics is higher for PMMA-adherent bacteria with respect to the conventional antibiotics compared to the silicone-elastomer adherent bacteria. Colonising bacteria are not significantly different in asymptomatic CL wearers concerning CFU and species: asymptomatic: CNS (54 vs. 47 %), propioni- (43 vs. 48 %), coryne (6 vs. 10 %) and gram-negative bacteria (2 vs. 3 %), S. aureus (2 vs. 1 %), and in patients with keratitis are not significantly different concerning species: CLARE (H. influenzae, S. pneumoniae), CLPU (S. aureus, S. pneumoniae), IK (P. aeruginosa), however, concerning the CFU (gram-negative) 23.7 % (keratitis) vs. 3.8 %, (asymptomatic), S. pneumoniae 7.6 % vs. 0.6 %. The epithelial expression of bacterial ligands like WGA is lower for HOTL (3230 p/750 micro) than for low oxygen-transmissible (4611 p/750 micro) consistent with a decreased adherence of pseudomonas of 5.31 - 5.98 vs. 5.9 - 7.81 bacteria/cell. Asymptomatic morphological anomalies and keratitis are less frequent in HOTL. Apoptosis induced desquamation and the central epithelial thickness decreased. In a one year study of 504 patients wearing continuous wear HOTL for 30 days, corneal staining was seen in 10.6 vs. 10.5 % 30-day 7 day extended wear, infiltrates in 2.3 vs. 4.6 %, epithelial oedema in 3.6 vs. 2.0 % und tarsal conjunctival anomalies in 3.0 vs. 1.6 %. Individual reports on infectious keratitis in HOTL wearers have been published. CONCLUSIONS: Fewer morphological anomalies and a decreased incidence of infections (on the basis of duration of wear) can be associated with the higher oxygen transmission of HOTL. From the clinical point of view, colonisation of HOTL with the same bacterial species as in conventional biomaterials, and the extended wear demand clear patient counseling and vigilant follow-up by an ophthalmologist. PMID- 12886504 TI - ["High-tech" in diagnostic procedures of the lacrimal system]. PMID- 12886505 TI - [Evidence of lacrimal plugs via high resolution ultrasound]. AB - BACKGROUND: The practical value of high-frequency ultrasound (transducer frequency of 20 MHz) for studying lacrimal plugs positioned into canaliculi was proved. MATERIALS AND METHODS: Twelve patients with twenty intracanalicular plugs and two punctum plugs were examined via high-frequency B-scan ultrasonography using 20 MHz transducer (model I3 Sacramento, USA). Detection and localisation of the intracanalicular plugs was made by a 20 MHz sector scanner. The ultrasound examinations were performed 1 - 24 month after the placement of lacrimal plugs. After patient's head positioning, the high-frequency ultrasound investigation was done via immersion fluid (2 % methylcellulose). RESULTS: All patients with dry eye treated by lacrimal plug implant showed echographic structure in the lacrimal canaliculus. In transversal echograms it was possible to image both canaliculi together when the lids were half-closed. Contrary to the normal state, it was not necessary to inject viscous fluid into the canaliculus. High-resolution ultrasound was able to differentiate the normal canaliculus from the findings after plug placement. The echograms can vary from one plug type to another. Highly reflective structures were found after the placement of silicone intracanalicular plugs, e. g. HERRICK-Plug. In contrast, the ultrasonic image taken through acrylic polymer intracanalicular plugs showed homogeneous small reflective inner structure, e. g. SMART-Plug. However, smooth and flat acoustic interface between acrylic polymer plug and the lacrimal canaliculus produced strong echoes. CONCLUSIONS: 20 MHz ultrasound seems to be well suited for the detection and localisation of intracanalicular plugs. By use of 20 MHz ultrasound scans it is possible to get high-quality images of the intracanalicular plug and around lacrimal canaliculus. Compared with UBM, the depth of penetration is much higher with negligible resolution. On the whole, we believe that 20 MHz ultrasound can become a useful tool for evaluating the placement of intracanalicular plugs after insertion. PMID- 12886506 TI - [Block excision of epithelial ingrowth after cataract surgery--report on 15 patients]. AB - BACKGROUND: Cystic or diffuse epithelial ingrowth into the anterior chamber after cataract surgery is a rare complication. It can lead to painful secondary glaucomas or in case of fistulation to persisting ocular hypotony or atrophy of the globe due to wrong or inadequate therapy. PATIENTS AND METHODS: The cause of epithelial ingrowth was a previous cataract surgery in 15 of 59 patients (25 %, Erlangen Block-Excision Registry for epithelial ingrowth). Eleven patients were females. Mean age was 65 +/- 13 years. RESULTS: Ten patients underwent block excision of epithelium, adjacent iris, ciliary body, sclera or cornea due to epithelial ingrowth following intracapsular cataract extraction between 1980 and 1987 and five patients since 1987 because of epithelial ingrowth following extracapsular cataract extraction (4 with and 1 without intraocular lens implantation); all primary cataract surgeries had been performed in external hospitals. We found a diffuse invasion in 3 eyes (all following icCE) and a cystic epithelial invasion in 12 eyes histologically. Mean time interval between cataract surgery and block excision was 7 +/- 8 years (range: 1 to 33 years). The excisional defect in the globes wall was covered using a tectonic corneoscleral graft (diameter: 8.0 +/- 1.8 mm). Postoperative astigmatism was 3.6 +/- 3.4 dpt. Six eyes had postoperatively a visual acuity of 20/200 or better. No recurrence of epithelial downgrowth was observed, nor was an enucleation needed. CONCLUSION: Surgical procedures with opening of the cyst (e.g. laser) are contraindicated and may lead to a transformation from cystic into diffuse epithelial invasion with potential blindness. The therapy of first choice in eyes with cystic epithelial ingrowth and an extension less than five clock hours is the curative block excision technique combined with tectonic corneoscleral graft. Surgical results are excellent in order to regard a curative excision and the survival of the eye with acceptable achieved function in consideration of the difficult primary situation with intra- and postoperative complications due to previous cataract surgery. PMID- 12886507 TI - [Indication and effect of immunosuppression in patients with scleritis]. AB - PURPOSE: The value of immunosuppressive drugs for the therapy of scleritis patients is unclear. The authors investigated the indications and effects of immunosuppression in a group of patients with scleral inflammation. METHOD: Retrospective study of patients treated for scleritis (n = 87) or episcleritis (n = 18). The demographic factors, clinical symptoms, visual outcome, course of inflammation, ocular complications resulting from inflammation, topical and systemic antiinflammatory medication, and associated systemic diseases were analysed. RESULTS: Only one patient with episcleritis, but 37 with scleritis presented with ocular complications (P = 0.003). The vision was impaired in 15 patients with scleritis, but not in episcleritis patients (P = 0.022). In the group of patients with episcleritis, only those with frequent relapses required more than topical antiinflammatory drugs, especially systemic non-steroidals. In contrast, systemic therapy was indicated in all of the scleritis patients. Ocular complications were found more often in patients with necrotising (n = 7/10) or posterior scleritis (n = 10/11) than in those with diffuse (9/39) or nodular (11/27) scleritis. Compared with the other patients, associated systemic autoimmune diseases were more common in patients with necrotising scleritis (P = 0.03). The need for immunosuppression was associated with vision-threatening complications (glaucoma, uveitis, peripheral ulcerative keratitis) (P < 0.01), systemic autoimmune disease, and necrotising and posterior form of scleritis (P < 0.01). Quiescence of scleritis was obtained in 59 of the scleritis patients, and improvement of inflammation was achieved in further 21. In 26 patients, scleritis did not improve with systemic steroid or non-steroidal treatment, but with immunosuppression. CONCLUSIONS: Scleritis is often associated with life threatening systemic diseases and vision-threatening ocular complications. In patients with severe scleritis, especially with the posterior and necrotising form, improvement can often be achieved with immunosuppression. PMID- 12886508 TI - [Post-traumatic endophthalmitis after penetrating injury in Vietnam: risk factors, microbiological aspect and visual outcome]. AB - BACKGROUND: Post-traumatic endophthalmitis associated with penetrating injury represents a distinct kind of intraocular infection and is still a severe problem for diagnosis and treatment in developing countries. We report the incidence, microbiological presentation and visual outcome of post-traumatic endophthalmitis in South Vietnam. PATIENTS AND METHOD: In a retrospective study medical reports of 515 patients with penetrating injury treated at Eye Hospital in Ho Chi Minh city, Vietnam, from 1/1999 to 5/2000 were reviewed and analysed. The treatment was standardised: 1. after vitreous-biopsy intravitreous injection of 0.1 mg/0.1 ml Gentamycine and 1 mg/0.1 ml Vancomycine; 2. subconjunctival injection of 25 mg/0.5 ml Vancomycine and 20 mg/0.5 ml Gentamycine; 3. topical and systemic antibiotic treatment due to microbiological presentation; 4. systemic steroids treatment. Clinical parameters were evaluated for association with an increased risk of endophthalmitis. RESULTS: Out of 515 patients with penetrating injury 61 patients (11.8 %) with endophthalmitis were treated and evaluated. Mean time interval from trauma to diagnosis of endophthalmitis was 16.8 +/- 5.6 days. Follow-up period was 15.6 +/- 23.4 days. Visual acuity at the diagnosis of endophthalmitis in 96 % of all patients was only finger counting or light perception. Enucleation was needed in 14 % of patients, 29 % of patients developed phthisis bulbi. A visual acuity better than finger counting was reached in only 10 % of all patients with endophthalmitis. Cultures of the vitreous aspirate were positive in only 50 % of cases available and showed Gram-positive (51 %), Gram-negative bacteria (33 %) and fungi (16 %). Risk factors found to be significant were: 1. a purely corneal wound; 2. wound length less than 5 mm; 3. surgical primary repair more than 24 hour after trauma; 4. inadequate antibiotic treatment and 5. a rural injury setting. CONCLUSION: Post-traumatic endophthalmitis is still a major problem in Vietnam. Delay in primary wound closure or inadequate antibiotic treatment worsen the prognosis profoundly. Comprehensive prophylactic antibiotic treatment at the time of injury repair is in dispensable. In risk eyes particular attention should be paid to prophylaxis and signs of infection. Further prospective studies are necessary to develop optimal and adequate diagnostic and treatment options in Vietnam. PMID- 12886509 TI - [Resource utilisation and cost of amblyopia treatment]. AB - BACKGROUND: The cost-effectiveness of screening for amblyopia is a controversial issue of international debate. The purpose of this study was to estimate the cost of amblyopia treatment to be used as a component for modelling the cost effectiveness of prevention programmes. Cost was calculated from the perspective of the German social health insurance in the year 2002. MATERIALS AND METHODS: A standardised detailed survey was conducted in writing among 13 experienced experts in amblyopia treatment from various offices and strabismological units in Germany. Average volumes of treatment items were estimated for a maximum treatment period of nine years. Cost was calculated using administrative prices (based on the social health insurance's uniform fee schedule for physician services and reference prices for therapeutic aids) and market prices. RESULTS: The questionnaires were fully completed by 12 of the 13 experts. The mean total cost of treatment was estimated at 2.472 Euro (95 %-CI: 1.171 - 3.774) for strabismic amblyopia and 2.051 Euro (95 %-CI: 426 - 3.675) for amblyopia of refractive origin. About 70 % of the total cost was caused by the therapeutic aids (e. g. glasses, patches). The price of the patches had a marked impact on the total treatment cost. CONCLUSIONS: The results may be used for modelling the cost-effectiveness of screening programmes for the prevention of amblyopia. PMID- 12886510 TI - [Choroidal neovascularisation in type II membranoproliferative glomerulonephritis, photodynamic therapy as a treatment option--a case report]. AB - BACKGROUND: Photodynamic therapy (PDT) is an established treatment in predominantly classic choroidal neovascularisation (CNV) in exudative age-related macular degeneration (AMD). Membranoproliferative glomerulonephritis (MPGN) type II leads to AMD-similar changes ("pseudo-AMD") of the fundus with CNV as a late complication. We report on a patient suffering from MPGN II and a classic CNV who was treated with PDT. PATIENT AND METHOD: A 31-year old male patient suffering from MPGN II from the age of 8 presented with visual deterioration and metamorphopsies on his left eye (VA OD 1.0, OS 0.25). Funduscopy revealed a "Pseudo"-AMD on both eyes and the left eye showed a 100 % classic CNV in fluorescein angiography (FLA). The left eye has treated with PDT. Visual acuity after PDT remained stable with 0.25 OS and metamorphopsies were reduced. 3 months later FLA showed a small residual leakage, therefore a second PDT was performed. 4 months after the 2 nd PDT the patient suffered again from metamorphopsies with a stable VA of 0.25 OS following a 3 rd PDT. RESULTS: Visual acuity of the left eye remained stable at 0.25 and metamorphopsies reduced due to the treatment of 3 PDT's. Even in "pseudo-AMD" in MPGN II the PDT is able to reduce leakage and stops progression consistent to the results in exudative AMD. CONCLUSION: In MPGN II associated "pseudo"-AMD with classic CNV the PDT was also successful. The origin of the CNV does not appear to play a role for the success of the PDT. PMID- 12886511 TI - [Complications following Molteno implant surgery: case report]. AB - We present a 43-year old patient with refractory secondary glaucoma, who showed epithelial downgrowth into the anterior chamber following Molteno implant surgery. It will be discussed, whether the Molteno implant or the following keratoplasty, which was necessary because of corneal decompensation, caused the epithelial downgrowth. PMID- 12886512 TI - [Ocular manifestation in LCAT deficiency--a clinicopathological correlation]. AB - BACKGROUND: Bilateral stromal corneal opacifications are important to detect potentially associated systemic diseases. Differential diagnosis includes, besides inflammatory diseases mucopolysaccharidoses, HDL deficiencies (LCAT deficiency, Tangier disease and fish eye disease), Schnyder's crystalline corneal dystrophy, lipid keratopathy, cystinosis, gout and mucolipidoses. CASE REPORT: The cornea of a 60-year old male patient, 180 cm height, showed milky hazy diffuse stromal inclusions with a prominent lipoid arc (VA O. D. 20/30 and O. S. 20/40, contre-jour VA less than 20/150). Ten years earlier, lattice corneal dystrophy was diagnosed. The cornea was thickened (0.61 mm). Except for the cornea, the anterior and posterior segments of the eye were unremarkable. Serum HDL concentration was decreased pathologically (7 mg/dl), although the LCAT activity was combined with apo-AI deficiency. Due to membrane instability, the erythrocytes showed target cell configuration. Penetrating excimer-laser keratoplasty was performed O. S. The epithelial wound closed only on the 32nd postoperative day. Histologically, the corneal stroma showed multiple vacuoles and amyloid deposits. Genetic analysis revealed two mutations in chromosome 16. CONCLUSION: To our knowledge, this is the second description in the literature of a patient with LCAT deficiency and secondary amyloidosis of the cornea. Additionally, LCAT deficiency is associated with anaemia and proteinuria. After a penetrating keratoplasty, prolonged wound healing is possible. Because of the bilateral corneal clouding, the ophthalmologist may help to identify patients with LCAT deficiency. Thus, it is possible to start antiarteriosclerotic therapy. PMID- 12886513 TI - A pheromonotropic peptide of Helicoverpa zea, with melanizing activity, interaction with PBAN, and distribution of immunoreactivity. AB - The sequence of an 18-amino acid residue peptide was deduced from the gene encoding PBAN and other peptides with common C-termini in Helicoverpa zea. The peptide caused melanization in larvae and pheromone production in females of H. zea, and was designated pheromonotropic melanizing peptide (Hez-PMP). The peptide has a 83% sequence homology with a pheromonotropic peptide isolated from Pseudaletia separata. PMP caused melanization and mortality when injected into larvae just before molting. Whereas intense melanization was caused with a dose of 1,000 pmol, peak mortality occurred at 100 pmol, with 50% of larvae dying within 48 h after injection. Pheromonotropic activity of PMP was dose dependent. Co-injection of Hez-PMP and Hez-PBAN into a female resulted in suppression of the pheromonotropic effect of PBAN. Whole-mount immunocytochemical studies revealed PMP-like immunoreactivity in frontal ganglion, subesophageal, thoracic, and abdominal ganglia as well as the esophageal nerve. PMID- 12886514 TI - Kinases, intracellular calcium, and apolipophorin-III influence the adhesion of larval hemocytes of the lepidopterous insect, Galleria mellonella. AB - Based on the results from the use of selective inhibitors and activators, active protein kinase A, protein tyrosine kinase, and protein kinase C (PKC) isoforms decreased the adhesion of larval Galleria mellonella hemocytes to glass slides. The protein kinase A inhibitor at all concentrations increased granular cell adhesion only whereas protein tyrosine kinase elevated both granular and plasmatocyte attachment at the lowest concentration. Active, Ca(2+)- and lipid dependent PKC isoforms limited plasmatocyte and granular cell adhesion whereas PKC that was inhibited by selected compounds (with differed modes of PKC inhibition) enhanced hemocyte attachment. The granular cells were more sensitive to the PKC inhibitors than were plasmatocytes. Phospholipase C and its diacylglyceride product were necessary to reduce hemocyte adhesion and maintain PKC activity. Extracellular Ca(2+), possibly transported through L-channels, was required for plasmatocyte attachment. In contrast, lowering the levels of cytosolic Ca(2+) was associated with decreased PKC activity and was required for hemocyte adhesion. PMID- 12886515 TI - Antisense inhibition of neuronal nicotinic receptors in the tobacco-feeding insect, Manduca sexta. AB - Acetylcholine is the predominant excitatory transmitter in the insect central nervous system with many of its effects mediated by nicotinic acetylcholine receptors. These receptors are present at very high density and are structurally heterogeneous, although little is known about functional distinctions between them. An interesting system for examining these receptors is the larval stage of Manduca sexta, a nicotine-resistant tobacco-feeding insect. The nicotinic responses of cultured neurons were found to be blocked by mecamylamine and curare but highly resistant to alpha-bungarotoxin. The responses were also unaffected by the reducing agent dithiothreitol and the alkylating agent bromoacetylcholine suggesting that the alpha-subunit dicysteine agonist binding site is protected. To begin determining the functional roles of different subunits in these receptors, cultured neurons were treated with oligonucleotides based on the gene sequence of the alpha subunit, MARA1. Antisense DNA caused a significant downward shift in the amplitude distribution of nicotinic responses compared to sense or reverse antisense treatments. These treatments did not affect currents mediated by the application of GABA. The reduction in the nicotinic depolarization and inward currents did not affect the rate of current onset or recovery, suggesting that antisense MARA1 causes a partial block of all nicotinic responses in these neurons. These results demonstrate that receptor gene expression in insect neurons can be manipulated in a sequence-specific manner by antisense treatment and they provide evidence that MARA1 is important for normal nicotinic responses in Manduca. PMID- 12886516 TI - Stem cells from midguts of Lepidopteran larvae: clues to the regulation of stem cell fate. AB - Previously, we showed that isolated stem cells from midguts of Heliothis virescens can be induced to multiply in response to a multiplication protein (MP) isolated from pupal fat body, or to differentiate to larval types of mature midgut cells in response to either of 4 differentiation factors (MDFs) isolated from larval midgut cell-conditioned medium or pupal hemolymph. In this work, we show that the responses to MDF-2 and MP in H. virescens stem cells decayed at different time intervals, implying that the receptors or response cascades for stem cell differentiation and multiplication may be different. However, the processes appeared to be linked, since conditioned medium and MDF-2 prevented the action of MP on stem cells; MP by itself appeared to repress stem cell differentiation. Epidermal growth factor, retinoic acid, and platelet-derived growth factor induced isolated midgut stem cells of H. virescens and Lymantria dispar to multiply and to differentiate to mature midgut cells characteristic of prepupal, pupal, and adult lepidopteran midgut epithelium, and to squamous-like cells and scales not characteristic of midgut tissue instead of the larval types of mature midgut epithelium induced by the MDFs. Midgut stem cells appear to be multipotent and their various differentiated fates can be influenced by several growth factors. PMID- 12886517 TI - In vivo preservation of steroid specificity in CWR22 xenografts having a mutated androgen receptor. AB - BACKGROUND: In vitro studies of CWR22 tumor cells lack steroid specificity. We sought to determine if CWR22 xenografts also lack steroid specificity. METHODS: We injected castrated nude mice with CWR22 tumor cells (6 x 10(6) cells) and implanted Alzet osmotic pumps that delivered approximately 1 mg steroid/kg body weight/day. RESULTS: Serum PSA levels were detectable in intact mice and castrated mice treated with testosterone (T), but not in those treated with estradiol (E(2)), progesterone (P), or flutamide (F). T maintained mean tumor weight similar to that in intact mice (P = NS). We observed no tumors in castrated mice or mice treated with E(2), P, or F, and tumor histology was consistent with weights. CONCLUSIONS: The mutation of the androgen receptor (H874Y) that occurs in the CWR22 xenograft model of human prostate cancer does not significantly affect in vivo steroid specificity for E(2), P, or F. PMID- 12886518 TI - Possibility of re-screening intervals of more than one year in men with PSA levels of 4.0 ng/ml or less. AB - BACKGROUND: The optimal re-screening interval is one of the most important issues to evaluate the effectiveness of screening for prostate cancer. METHODS: Between 1992 and 2000, 7,026 men aged 50-78 with baseline PSA levels of 4.0 ng/ml or lower underwent screening for prostate cancer twice or more. The risk of developing prostate cancer relative to elapsed years and baseline PSA levels were investigated. RESULTS: Prostate cancer was detected in a total of 127 cases (1.8%). The detection rate of prostate cancer was high between 1.6% and 5.5% at 1 year after baseline PSA measurements in men with baseline PSA levels of 2.1-4.0 ng/ml. In men with baseline PSA levels of 1.1- 2.0 ng/ml, the detection rate increased from 0.06% to 1.02% with passed years. The proportion of stage >/=T3 was high at 63% in prostate cancer cases detected between 3 and 4 years after baseline PSA levels being 1.1-2.0 ng/ml. In men with baseline PSA levels of 1.0 or lower, the cumulative detection rate of prostate cancer was low at 0.01% within 3 years, however, the detection rate increased to 0.34% after 5 or more years from baseline PSA measurements. CONCLUSIONS: The re-screening interval was recommended to be 1, 1-2, and 3-5 years for men with baseline PSA levels of 2.1 4.0 ng/ml, 1.1-2.0 ng/ml, and 1.0 ng/ml or lower, respectively. PMID- 12886519 TI - Perillyl alcohol mediated radiosensitization via augmentation of the Fas pathway in prostate cancer cells. AB - BACKGROUND: The management of hormone-insensitive locally advanced prostate cancer is difficult and complex and there is an urgent need for the development of effective chemotherapeutic agents intended for combination with currently available treatment modalities. METHODS: The present paper demonstrates the effectiveness of the monoterpene perillyl alcohol (POH) as potent radiosensitizer on DU145 and PC3 cell lines by performing clonogenic survival assays, cycle analysis, and assays to detect viability, apoptosis, and Fas receptor/ligand by flow cytometry. RESULTS: POH pretreatment resulted in a dose dependent sensitization to kill cell by radiation. Furthermore, POH treatment induced a transient G(2)/M arrest, enhanced the expression of the membrane bound form of the Fas ligand and sensitized the cells to Fas mediated apoptosis. CONCLUSIONS: The unique manner of radiosensitization in addition to its low toxicity profile makes POH a promising new agent for preclinical evaluation as a potential radiosensitizer in the treatment of prostate cancer. PMID- 12886520 TI - Apoptosis levels increase after castration in the CWR22 human prostate cancer xenograft. AB - BACKGROUND: The role of apoptosis in the regression of human prostate cancer after androgen deprivation therapy remains controversial. Detection of caspase-3, an ubiquitous effector of apoptosis, is a highly specific technique for in vivo evaluation of apoptosis. METHODS: Apoptotic rates were evaluated in the androgen dependent CWR22 human prostate cancer xenograft in tumors that represented time points throughout the progression from androgen-stimulated to recurrent prostate cancer. Caspase-3 levels in formalin-fixed, paraffin-embedded specimens were quantified using immunohistochemical detection and video image analysis. Western blot analysis was used to confirm the results of immunodetection. RESULTS: Expression of caspase-3 reached a maximum on day 2 after castration, decreased on day 6, and remained low until tumor recurrence. The percentage of tumor area expressing caspase-3 increased from 2.51 +/- 0.44% in tumors from intact mice to 20.84 +/- 1.75% on day 2 after castration. Among immunopositive cells, the intensity of caspase-3 expression measured using the mean optical density (MOD) increased 45% (0.3762 +/- 0.003 to 0.5461 +/- 0.001) on day 2 after castration compared to levels detected in tumors from intact mice. CONCLUSIONS: Apoptosis contributes to tumor regression after castration in the CWR22 human prostate cancer xenograft model. PMID- 12886521 TI - Foundation for the integration of biologically based therapy in the management of prostate cancer. AB - The cure of advanced prostate cancer (PCa) will be achieved through the clinical application of biological observations. To achieve this, a new classification of human PCa is needed. Unlike in previous classifications of PCa which were based on anatomic determinates of outcome, we propose to integrate emerging PCa biology into this taxonomy. Three biologically based categories were identified (amphotropic, clonal expansion, and heterotopic), and each has unique features that influence therapy selection. It is unlikely that any single agent or modality will successfully control PCa. It is hoped that the classification and trial designs we propose will facilitate the application of integrated treatment and prevention strategies. PMID- 12886522 TI - Extensive analysis of the 13q14 region in human prostate tumors: DNA analysis and quantitative expression of genes lying in the interval of deletion. AB - BACKGROUND: Loss of heterozygosity (LOH) on chromosome arm 13q14 is one of the most consistent genetic alterations in sporadic prostate cancer. This alteration may be involved in prostate oncogenesis through inactivation of one or more tumor suppressor genes (TSGs). Candidate gene expression is an approach to focus the search for TSGs in this region. METHODS: We tested 41 human sporadic prostate tumors for 13q14 LOH by using seven polymorphic markers overlapping the critical region and used a real-time quantitative RT-PCR assay to study the same tumors for expression of the 31 genes located in this genomic region (localized by the Human Genome Project Working Draft). RESULTS: Allelic loss on at least one locus was found in 18 (41%) of the 41 tumor DNAs. Only four genes (ITM2B, CHC1L, KIAA0970, and LOC51131), located in the region most frequently deleted in prostate carcinoma, showed a significant difference in expression between normal and neoplastic prostate tissues. CONCLUSIONS: Given their location in the LOH hotspot, as indicated by our genomic analysis, ITM2B, CHC1L, KIAA0970, and LOC51131 are candidate tumor suppressor genes in this region. ITM2B that showed a significant association (P < 0.005) between expression and LOH at the corresponding locus could, furthermore, be the main target of the observed LOH at 13q in prostate tumors. PMID- 12886523 TI - Androgen-induced norepinephrine release mediating guinea pig seminal vesicle smooth muscle proliferation: potential role of pre-synaptic alpha2-adrenoceptors. AB - BACKGROUND: Recent studies from our laboratory have demonstrated that androgen induced basal norepinephrine (NE) release is responsible for the onset of proliferation in seminal vesicle smooth muscle (SVM) cells during early puberty. With subsequent sexual maturation, SVM was irreversibly differentiated to an androgen-resistant-amitotic state in which basal NE release remained elevated and resistant to androgen withdrawal or repletion. Based on these findings, we hypothesized that this irreversible elevation of basal NE release during pubertal development is caused, at least in part, by the down-regulation of pre-synaptic NE feedback inhibition, secondary to irreversible reduction in the expression of neuronal (pre-synaptic) alpha(2)-adrenoceptors. Functional alpha(2)-adrenoceptors are selectively localized to pre-synaptic sites in SVM. METHODS: To test this hypothesis, we employed ligand binding techniques with [(3)H]RX821002, an antagonist which labeled all alpha(2)-adrenoceptor sub-types. Initial experiments focused on analysis of competitor specificity to identify the predominant alpha(2)-adrenoceptor sub-type in SVM. Subsequently, we quantified the changes in the receptor concentration (B(max)) for [(3)H]RX821002 at the point of maximal dihydrotestosterone (DHT)-induced change in basal NE release. RESULTS: Based on competitor specificity for [(3)H]RX821002, the alpha(2D)-adrenoceptor sub-type predominated in SVM. We treated pre-pubertal castrate animals with DHT for 7 days, which was previously demonstrated to maximally induce basal NE release. This treatment reduced the pre-synaptic alpha(2)-adrenoceptor B(max) 4-fold. In animals which had been castrated as adults, the B(max) for [(3)H]RX821002 remained irreversibly suppressed. CONCLUSIONS: The DHT-dependent reduction in the alpha(2)-adrenoceptor concentration is consistent with the developmental pattern of increased basal NE release. These findings support the hypothesis that the down-regulation of pre-synaptic NE feedback is mechanistically involved in the irreversible elevation of basal NE release. NE mediates proliferation in SVM in early pubertal development. Thus, the androgen-dependent pubertal growth of smooth muscle cells may be indirectly controlled at the level of neurotransmission. PMID- 12886524 TI - Increased growth factor production in a human prostatic stromal cell culture model caused by hypoxia. AB - BACKGROUND: Local hypoxia may be one of the triggers of embryonic reawakening of the stroma and subsequent hyperplastic growth in the prostate. Using a cell culture model of human prostatic stromal cells, we investigated the effects of hypoxia on activation of hypoxia-inducible factor 1 (HIF 1) and on the production of growth factors. METHODS: Primary prostatic stromal cells were grown in normal and hypoxic (1% O(2)) atmosphere. Activation of HIF 1 was evaluated after different time intervals by Western blot. Induced secretion of growth factors VEGF, FGF-7, TGF-beta, IL 8, and FGF-2 were analyzed by ELISA. To confirm the in vitro findings we also performed immunohistochemistry of HIF 1alpha as well as pro-collagen I, collagens I, III, and IV in the benign tissue of radical prostatectomy specimens. RESULTS: HIF 1 is activated in a time-dependent manner, already starting 1 hr after exposure of stromal cells to hypoxic conditions. Secretion of VEGF, FGF-7, TGF-beta, FGF-2, and IL 8 is increased under hypoxic in vitro conditions in comparison to normoxia. Levels of TGF-beta, VEGF, and IL 8 were rapidly and statistically significantly increased in the supernatant of hypoxic cells. Consistent with the in vitro findings, immunohistochemistry of HIF 1alpha in (benign prostatic hyperplasia) BPH tissue revealed strong HIF 1alpha nuclear staining in hyperplastic areas. No difference was observed in the collagen pattern between hyperplastic and normal prostate tissue. CONCLUSIONS: Prostatic stromal cells respond to hypoxia by upregulation of secretion of several growth factors suggesting that hypoxia can trigger prostatic growth. Therefore, hypoxia might be a key factor contributing to the pathogenesis of BPH. PMID- 12886525 TI - C16 ceramide accumulates following androgen ablation in LNCaP prostate cancer cells. AB - BACKGROUND: Adenocarcinoma of the prostate is the most frequently diagnosed non cutaneous cancer and the second leading cause of cancer-related deaths among men in the United States. The most successful therapies to date for this tumor have involved some form of androgen ablation. However, these therapies become ineffective as the tumor evolves to an androgen-insensitive state. Ceramide is a lipid second messenger that has been shown to mediate growth arrest or cell death when added exogenously to prostate cancer cells. As a first step toward understanding the events that lead to the transition of prostate cancer cells to an androgen-independent state, we considered investigating the effect of androgen ablation on endogenous ceramide levels in androgen-sensitive and androgen insensitive prostate cancer cells. METHODS: To investigate the mechanisms of growth arrest/apoptosis in androgen-sensitive (LNCaP) and insensitive (DU-145, PC 3) cells, we used various methods including nonyl acridine orange (NAO) staining, propidium iodide (PI) staining/cell-cycle analysis, lipid analysis, and Western blotting assays. RESULTS: In this study, we demonstrate that androgen ablation drives G(0)/G(1)-phase cell-cycle arrest followed by progressive apoptosis in vitro, in LNCaP cells. Lipid analysis indicated an increase in C16 ceramide, which was generated via the de novo pathway as revealed by blockade of ceramide synthase by fumonisin B1. The addition of 5alpha-dihydrotestosterone (DHT) or fumonisin B1 rescued LNCaP cells from apoptosis induced by androgen ablation, and decreased levels of intracellular C16 ceramide. Neither apoptosis nor an increase in C16 ceramide was observed in androgen-independent cell lines following androgen ablation. PMID- 12886526 TI - Induction of cellular and humoral immune responses to tumor cells and peptides in HLA-A24 positive hormone-refractory prostate cancer patients by peptide vaccination. AB - BACKGROUND: To assess the safety and immune response of a peptide-based immunotherapy for patients with hormone-refractory prostate cancer, a phase I clinical trial was conducted. METHODS: This study first investigated whether cytotoxic T-lymphocyte (CTL) precursors reacting to peptide with vaccine candidates (14 peptides for HLA-A24 positive patients) were detectable in the pre vaccination peripheral blood mononuclear cells (PBMCs) of ten patients with hormone-refractory prostate cancer. Patients were then vaccinated subcutaneously with only those peptides to which pre-vaccination PBMCs reacted (CTL precursor oriented peptide vaccine) for up to four kinds of peptides. RESULTS: Overall vaccinations were generally well tolerated, but most patients (nine of ten) developed grade 1 local redness and swelling at the injection site. Increased CTL response to both peptides and cancer cells were observed in four of ten patients. Anti-peptide IgG antibodies were also detected in post-vaccination sera of seven of ten patients. One patient achieved a partial response with an 89% decrease in PSA. Stable disease was demonstrated in five of ten patients (50%) for the median duration of 2 months (range, 2-5 months). There were no objective responses of measurable lesions. CONCLUSIONS: Increase in cellular and humoral immune responses, and decrease in PSA level in some patients support further development of peptide-based immunotherapy for hormone refractory prostate cancer. PMID- 12886527 TI - [The ways of improvement of surgeons' training at the Department of Managerial Medical Staff of the Military Medicine Academy]. AB - The urgent need to discuss with military surgeons the problem of improvement of educational process in the faculty of managerial medical staff of the Military and-Medical Academy under the new social-and-economic conditions against the background of the RF Armed Forces reformation has prompted the authors to write this article. The first report deals with the problems of general organization of educational process taking into account the needs for surgeons with higher military education and creation of certified specialized surgical care in the Troops. The authors suggest that all people with interests at stake should express their opinion concerning this problem. PMID- 12886528 TI - [Initial stage of development of military medical service in Russia (the tricentennial of the Armed Forces Supply System)]. PMID- 12886529 TI - [Medical expert evaluation of outcomes of extremity wounds]. PMID- 12886530 TI - [Organization of primary specialization (internship) of military pharmacists at the Tomsk Military Medical Institute]. PMID- 12886531 TI - [The problem of prophylaxis of dioxin intoxication (review of the literature)]. PMID- 12886532 TI - [Surgical treatment of perforated gastroduodenal ulcers in young persons]. AB - The knowledge of surgical tactics in treatment of ulcerous disease and its complications in young persons is of great practical importance for military health service. The problem of choice of surgical method in perforated gastroduodenal ulcers remains contradictory. Many authors draw attention to merits and demerits of both palliative and radical interventions. The long-term results of identical operations obtained by different authors often vary and not infrequently are quite opposite. The review of our and foreign literature concerning the surgical treatment of perforated gastroduodenal ulcers in the young patients shows that the unequivocal decision of this problem today is not available. Discussions about expediency to perform different interventions in the young patients are generally concentrated around the notions of "acute" and "chronic" ulcer. However both these categories are poorly defined because the criteria of their evaluation vary in different authors. Obviously the choice of surgical method in the young patients with perforated gastroduodenal ulcers should be based both on multi-factor analysis of anamnestic data, the patient's general state, intraoperative picture and on retrospective and prospective analysis of conservative treatment, its possibilities and efficiency, on the patient's opinion. The availability of technical possibilities to perform the operation and presence of the adequately trained surgeons are necessary. PMID- 12886533 TI - [Therapeutic effects of thermal sources of military sanatorium "Paratunka"]. PMID- 12886534 TI - [Clinical peculiarities of asthenic and dysthymic neurosis-like disorders in patients with alcoholism]. PMID- 12886535 TI - [Lipid peroxidation processes in epidermis of patients with psoriasis]. AB - The evaluation of local peroxide status was studied in 42 patients with psoriasis depending on the disease stage in different age periods (from 18 to 67 years) with disease duration from 2 months to 20 years. It was established that the development of psoriatic process is accompanied with the marked changes in local peroxide status in the epidermis. It is manifested by the decrease in intensity of lipid free-and-radical oxidation. Decrease in the products of lipoperoxidation occurs in epidermis--both in the affected and intact areas in all investigated patients independently of the stage of psoriatic process. It is more expressed during the aggravation period. The minimal levels of the intermediators being studied are recorded in the psoriatic plaques. PMID- 12886536 TI - [Experience in the use of flixotide in the treatment of bronchial asthma]. PMID- 12886537 TI - [Baneocin -- a drug of choice in prophylaxis and treatment of bacterial infections of the skin]. PMID- 12886538 TI - [Surgical treatment of ulnar and pre-patellar bursitis under ambulatory conditions]. PMID- 12886539 TI - [Regular medical check-up on patients with Lyme disease]. PMID- 12886540 TI - [Low doses of radiation and functional state of the central nervous system]. PMID- 12886541 TI - [Psychological dysadaptation and its psychoprophylaxis in students of higher aviation schools (psychophysiologic aspect)]. PMID- 12886542 TI - [Morbidity in seamen during long voyages according to the results of long-term studies (Report II)]. PMID- 12886543 TI - [How to measure public health in Denmark?]. PMID- 12886544 TI - [More recent measures for a nation's state of health--a review]. AB - The objective of this article was to give a conceptual survey of old and new measures for a nation's state of health with special focus on new measures seeking to combine mortality with morbidity, functional, and quality of life dimensions. Internationally, the development has given rise to two different movements. One (represented by Denmark and the rest of the EU) aims to develop a large number of standardized indicators, whereas the other (represented by the WHO and the World Bank) builds on the idea that it must be possible to combine the many indicators into a single summary measure for a nation's state of health. In the summary measures, distinction is made between health expectancy and health gap measures. Attention is given to reviewing DALY (disability-adjusted life years), disability weighting, and the social value choices applied in WHO's World Health Report 2000. PMID- 12886545 TI - [Disseminated sclerosis and sexuality]. AB - Since the onset of multiple sclerosis (MS) mainly occurs in younger persons between the age of 20 and 40, sexual dysfunctions have great impact on their quality of life. About 50% of all female and about 75% of all male patients complain of sexual dysfunctions. The primary symptoms among males are erective and ejaculative dysfunctions and reduced libido, while female patients mainly complain of reduced libido, problems achieving orgasm, decreased vaginal lubrication and changes in vaginal sensitivity. Secondary organic symptoms include fatigue, spasticity, muscular weakness, bladder problems, pain, cognitive and behavioural changes. Tertiary dysfunctions refer to general psychosocial problems in relation to chronic, progressive disease. One third of all couples in which either the man or the woman suffers from MS complain of problems in sexual and marital life, where especially the healthy female partner in general has sexual problems. Diagnosing and treating sexual dysfunctions in MS should ideally be carried out by a specialized "MS-team" with the core professionals being the neurologist, urologist, (neuro) psychologist and the nurse. Information about symptoms and their possible causes is an important part of the treatment, and not least learning more efficient coping strategies. Both for the patient and for the couple honest and open informative communication including information about sexual aids and perhaps also medical treatment will often result in minimizing the sexual problems and increasing quality of life. PMID- 12886546 TI - [Cost-of-illness of asthma in Denmark in the year 2000]. AB - INTRODUCTION: The total number of asthmatic patients in Denmark in the year 2000 was about 300,000 out of a total population of 5.3 million. The aim of this study was to evaluate cost-of-illness for all asthma patients in Denmark for the year 2000. MATERIAL AND METHODS: Direct and indirect costs were obtained from two Danish studies, Sorensen et al [1] and Sondergaard et al [2]. These studies allowed calculation of direct and indirect costs per year per patient. The costs were estimated in 2000-prices. Estimated cost of drugs was extrapolated to the population of asthmatics for which the actual cost was available from nation-wide statistics. The estimated costs of drugs were compared with the actual costs. As costs of asthma increase with the severity of the disease, a correction factor was obtained from the ratio between the actual costs of drugs and the estimated costs. This factor was used to normalise the subpopulations to reflect the entire group of asthma patients. RESULTS: The total costs of asthma were estimated at 1.9 billion DKK (745 DKK = 100 EURO, year 2000), the direct costs to medicine and treatment were estimated at 1.1 billion DKK and the indirect costs to sick-leave and early retirement pension were estimated at 0.8 billion DKK. Due to the differences in the estimates given by the sources, especially on the indirect costs, we obtained a range of the total cost from 1.4 billion DKK to 2.9 billion DKK in a sensitivity analysis. DISCUSSION: The estimated cost is consistent with data from other national and international sources, suggesting that our estimate be close to the real cost. Costs due to asthma may be expected to increase due to the increase in prevalence of asthma. PMID- 12886547 TI - [Protein-losing enteropathy--a new alternative of diagnosing and monitoring]. AB - Protein-losing gastroenteropathy (PE) is characterised by excessive loss of protein into the gastrointestinal tract. This condition associates not only with gastrointestinal disorders, but also with heart diseases, collagenoses, and several other generalized disorders. The symptoms are multiform and not necessarily abdominal. The treatment may be causal or symptomatic, and quite frequently it is possible to reduce the protein loss. The pathogenesis is considered multifactorial, but our knowledge of PE, including prevalence and clinical significance, is sparse, partly due to lack of appropriate examination procedures. Previous techniques were cumbersome and hampered by shortcomings. With 111In-transferrin, quantification and scintigraphic localization of the protein loss are obtained in one process and faeces collection is no longer needed. The method is promising, but further evaluation is necessary before it is brought into common clinical use. PMID- 12886548 TI - [Immunosuppressive treatment of the myelodysplastic syndrome]. AB - INTRODUCTION: The myelodysplastic syndrome (MDS) is featured by cytopenia in one or more cell lineages. Until recently the only possible treatment was supportive care with transfusions and antibiotics, but new options have now become available. In this article the results of immunosuppressive treatment are described and discussed. MATERIAL AND METHODS: We present seven patients who were treated with antithymocyte globulin (ATG) and cyclosporine. Previous series of patients with MDS treated with ATG are summarized. RESULTS: Two of the seven patients treated with ATG and cyclosporine achieved a complete response, one had a partial response and one had a minor response. DISCUSSION: In MDS an acquired insult to the hemopoietic stem cell leads to impaired differentiation and myelodysplasia. A model of MDS pathophysiology suggests that transformation of normal stem cells induces an autoimmune T cell response with the bone marrow as the target organ. This autoimmune attack results in chronic overproduction of pro apoptotic cytokines, especially TNF-alpha. This may contribute to a dysplastic morphology and increased apoptosis in the marrow. It is suggested that the mechanism of action of ATG in MDS may involve elimination of CD8+ lymphocyte mediated suppression of granulocyte/monocyte progenitor cells. PMID- 12886549 TI - [The Steno-2 study. Intensive multifactorial intervention reduces the occurrence of cardiovascular disease in patients with type 2 diabetes]. PMID- 12886550 TI - ["Kiss the child"--removal of nasal foreign bodies in children using the mouth-to mouth method]. AB - Two cases are presented, in which nasal foreign bodies in children were removed by a positive pressure technique using the mouth-to-mouth method. The technique is efficient, simple to perform and atraumatic to the child. The technique is suggested as first choice treatment in small children with nasal foreign bodies. PMID- 12886551 TI - [Idiopathic giant cell myocarditis with fatal outcome in a 37-year-old man]. AB - A case of idiopathic giant cell myocarditis (IGM) causing symptoms of left ventricular failure in a previously healthy 37-year-old man is reported. Sixteen months after the occurrence of the symptoms the patient was admitted to hospital with intractable cardiac arrest. Until this point he had been treated with different medications for left ventricular failure. The disease was finally diagnosed postmortem. PMID- 12886552 TI - [Picture of the month: Cordylobia antropophaga]. PMID- 12886553 TI - [Who is to be treated with statin?]. PMID- 12886554 TI - [Doubt about the harmlessness of statins]. PMID- 12886555 TI - [Dishonesty in clinical trials is expensive for the society 1]. PMID- 12886556 TI - [Dishonesty in clinical trials is expensive for the society 2]. PMID- 12886557 TI - [Dishonesty in clinical trials is expensive for the society 3]. PMID- 12886558 TI - [Righteous war for peace and democracy]. PMID- 12886559 TI - [Were the wars in Iraq for democracy and peace?]. PMID- 12886560 TI - [The frames for treatment. Mentally ill Turks here and there]. PMID- 12886561 TI - [Geriatric rehabilitation potential--who has it?]. PMID- 12886562 TI - [Who should be admitted to a geriatric unit?]. AB - Potential geriatric patients are frail old persons characterized by the presence of one or several geriatric conditions: polypharmacy, socio-economic problems, vision/hearing impairment, confusion, impaired mobility, incontinence, weight loss, depression, falls, dementia, and prolonged bedrest. Admission of these frail old patients to a geriatric unit may reduce mortality, improve function, and decrease the risk of nursing home placement. These frail old patients may not all be able to profit from geriatric rehabilitation. The published randomized trials showing substantial effect of geriatric rehabilitation exclude patients with severe dementia, terminal disease or those with a single medical disease for which there is no further treatment. Careful selection of patients is necessary in order to achieve superior, clear-cut effect when compared to conventional treatment in general medical wards. PMID- 12886563 TI - [Alendronate once a week]. PMID- 12886564 TI - [Rectal cancer treated at a county hospital. A prospective study of an 11-year period]. AB - INTRODUCTION: The results of rectal cancer surgery in a county hospital during an 11-year period from 1991 to 2002 are presented. Complications, local recurrence rate and survival were registered. MATERIAL AND METHODS: One hundred patients operated by one surgeon were included in the study. Twelve were Dukes' type A, 36 B, 48 C, and four were D. Low anterior resection was performed in 68 patients (six with J-pouch), abdominoperineal resection in 29 and a Hartmann's procedure in three patients. The operation was radical in 80 patients. Until December 31, 1994 a conventional operation was performed in 34 patients. Thereafter total mesorectal excision was introduced with excision of the mesorectum for tumours below 10 cm from the anal verge and resection of the mesorectum 5 cm below tumour for higher tumours (46 patients). RESULTS: Thirty days mortality was 4%. The clinical leakage rate after resection was 10% and 6% ended up with a permanent stoma. There were no deaths among patients with leakage. As a consequence of the change in surgical technique from January 1, 1995 the 5-year local recurrence rate declined from 0.32 to 0.11 (p = 0.13) and the overall survival rate exhibited a slightly rising tendency from 0.62 to 0.70 (p = 0.47). DISCUSSION: Operations for rectal cancer can be performed in a county hospital with a satisfactory outcome as to complication rates and operative mortality provided the operations are concentrated on few hands. The local recurrence rate and the overall survival showed non-significant tendencies to improvement with the introduction of a more radical operation which is in accordance with results from international trials. PMID- 12886565 TI - [Is change of general practitioners more frequent among patients diagnosed with cancer or other serious diseases?]. AB - INTRODUCTION: Change of GP may imply that the patient is dissatisfied with the health service provided. The aim of this study was to investigate whether newly diagnosed cancer patients change their GP more often than other patient groups and the background population. MATERIAL AND METHODS: The investigation was conducted using data on inhabitants from the counties of Aarhus, Vejle and South Jutland. We included 900 Group 1 insured patients, aged 18-75 years, distributed into three groups: 1) frequent cancers, 2) rare cancers and 3) benign, but chronic or severe new diseases. The diagnoses included occurred for the first time at the time of inclusion. Three hundred controls were randomly selected from the regional health care registry in Aarhus. The study period was from March 1 to August 30 1994. A change of GP was defined as a change not caused by the patient's change of residence or the GP's change of practice registration number due to expansion, takeover or split up of practice. RESULTS: In the three disease groups, 28 cases of change of GP were registered with a frequency of 5% (frequent cancers), 2.7% (rare cancers) and 1.7% (benign, but chronic or severe new diseases). The frequency in the background population was 2.7%. None of the differences were statistically significant. DISCUSSION: During the first year following a cancer or chronic disease diagnosis, change of GP is rare, particularly among patients with rare cancers. We reject the hypothesis that change of GP is frequent among newly diagnosed cancer patients. PMID- 12886566 TI - [Respiratory syncytial virus infections in children in general practice]. AB - INTRODUCTION: The aim of the study was to describe the course of respiratory syncytial virus (RSV) infections in children under two years of age seen in general practice. MATERIAL AND METHODS: Children under two years of age presenting acute respiratory infection during the registration period on 59 GPs' lists participated in the study. The GPs recorded data on a registration chart and a questionnaire was sent to the parents of the children in question one month after the date of inclusion. The children were tested in general practice for the presence of RSV. The GPs' objective findings and choice of treatment as well as the parents' account of the course of disease were compared in children with and without the presence of RSV. A total of 221 children participated in the study. RESULTS: Fifty-seven children were found RSV positive (25.8%). Among the RSV positive children there were significantly more with wheezing audibly detected with examination by stethoscope than among the RSV negative. The remaining parameters (the GP's objective examination, treatment and course of the disease) were distributed independently of the result of the RSV analysis. DISCUSSION: The results showed that RSV infections in children under two years in general practice are frequent and that the clinical picture most often is uncomplicated. PMID- 12886567 TI - [Small bowel necrosis associated with postoperative percutaneous jejunal tube feeding]. AB - Small bowel necrosis is a rare but highly lethal complication associated with postoperative jejunal tube feeding. The causative mechanism remains unclear but is most likely the result of several factors. The ischemic necrosis was preceded by progressive abdominal pain, distension, hypotension and hypovolemia. Timely recognition and surgical intervention may save the patient's life. Percutaneous enteral tube feeding may be used with utmost caution. PMID- 12886568 TI - [Compartment syndrome of the lower part of the leg, exceptional trauma mechanism]. AB - Acute compartment syndrome (CS) is a limb threatening condition which warrants emergency treatment. We describe a case of a 37-year-old man with acute CS developed without major trauma. Early diagnosis and prompt treatment by decompressive fasciotomy is of vital importance in order to preserve limb function and to avoid complications. As a consequence of this case the department purchased a pressure transducer. Measurement of high pressure could have saved the patient for an undue risk of serious complications. PMID- 12886569 TI - [Picture of the month: incrusted urinary catheter]. PMID- 12886570 TI - [Increased number of the elderly, but fewer with functional disabilities]. PMID- 12886571 TI - [Will the needs of the new surgical technique for patients with Parkinson disease be covered?]. PMID- 12886572 TI - [Migraine--acupuncture a good alternative]. PMID- 12886573 TI - [The General Practitioners' Employers' Association is below the standards]. PMID- 12886574 TI - [The allowance administrators in the municipalities have made their own bed, so they must lie on it!]. PMID- 12886575 TI - [Research as bridge-building between specialties]. PMID- 12886576 TI - Forensic dentistry: insurance for a lasting impression. PMID- 12886577 TI - 58,228 children with disabilities in Massachusetts. AB - The 2000 Census data on the numbers of children with disabilities in Massachusetts are presented in each community in the state. There are numerous difficulties associated with the delivery of dental care to these youngsters with special needs. The 2002 Kids Count Census Data Online, developed by The Annie E. Casey Foundation (using Census Bureau 2000 data), provides an extended detailed numeric picture of noninstitutionalized United States children between the ages of 5 and 15 at the state, county, city, New England town, and metropolitan area levels. In addition to standard demographic characteristics, the report provides a detailed review of the prevalence of children with one or more disabilities (including long-term sensory, physical, mental, and self-care limitations. PMID- 12886578 TI - Gastroesophageal reflux in children with cerebral palsy and its relationship to erosion of primary and permanent teeth. AB - Patients with cerebral palsy are known to have a high incidence of feeding difficulties, including problems with swallowing, vomiting, recurrent chest infections, and irritability. Gastroesophageal reflux (GER) is an involuntary passage of the gastric contents into the esophagus. It has been found in a higher prevalence (up to 75 percent) in cerebral palsy patients. Long-term gastric acid attacks teeth and can cause dental erosion. Bargen and Austin first discovered the relationship between GER and dental erosion in 1937 when they concluded that the loss of dental hard tissue could be an indicator and the predominant oral manifestation of GER. The purpose of this study is to investigate the correlation between GER and dental erosion in primary and permanent teeth in cerebral palsy patients. Twenty-one patients participated in this study. Results showed that out of the 21 patients, 15 had erosion and 11 of them had GER history. The chi-square analysis with the p value of less than or equal to 0.025 showed the distribution to be significant. The severity of the erosion was correlated to the duration of the disease, frequency of vomiting, pH of the acid, type of acid, and quantity and quality of saliva. The study revealed that children with cerebral palsy have an increased prevalence of tooth erosion, which may be attributed to the existing gastroesophageal reflux. PMID- 12886579 TI - Salute to Harvard School of Dental Medicine. PMID- 12886580 TI - Ultracet: a new combination analgesic. PMID- 12886581 TI - Idiopathic thrombocytopenic purpura presenting as gingival bleeding. PMID- 12886582 TI - A clinico-pathologic correlation. Epithelial and fibrous hyperplasia suggestive but not pathognomonic of condyloma acuminatum HPV infection. PMID- 12886583 TI - [Increase of lysosomal cationic proteins of polymorphonuclear leukocytes as a response to opisthorchis invasion]. AB - Cytochemistry was used to study the level of lysosomal cationic proteins of polymorphonuclear peripheral leukocytes in 25 and 20 patients with acute and chronic opisthorchiasis, respectively, before treatment. Comparison groups included patients with chronic viral hepatitis C, bronchial asthma, as well as apparently healthy individuals free of Opisthorchis invasion. That the ratios of an increase in the intracellular content of lysosomal cationic proteins to that in the control individuals were found to be 13.86 and 4.50 for acute and chronic opisthorchiasis, respectively. At the same time, the contribution of an allergic component (bronchial asthma) to the changes in the index studied is greater than that of somatic lesions of the hepatic parenchyma (viral hepatitis C). The authors suggest that it is expedient to use lysosomal cationic proteins in combination with other indices for making a diagnosis and determining the stage of the disease in Opisthorchis invasion. PMID- 12886584 TI - [Impact of anthelminthic therapy for opisthorchiasis on the biochemical parameters of hepatic function]. AB - A response of a complex of biochemical parameters of hepatic function to therapeutic exposures was studied at different stages of development of Opisthorchis invasion. The activity of AST, ALT, GGT, GGT, AP, AM, and CE and the concentration of bilirubin, cholesterol, and glucose in the sera from 232 patients with acute or chronic opisthorchiasis were studied. Their study was conducted in several steps: before treatment, after a course of pathogenetic therapy, 1-3 days after antihelminthic therapy with bilthricide and azinox, in early (following 1-3 months) and late (following more than 6 months) residual periods. Pathogenetic therapy was found to alleviate an acute inflammatory process in the liver, but without eliminating cholestasis completely. Antihelminthic therapy improved the results of hepatic tests in different periods after treatment for acute or chronic opisthorchiasis in relation to the rate of compensation of structural lesions. By and large, after pathogenetic and antihelminthic therapy for opisthorchiasis the parameters characterizing the cytolytic syndrome became more rapidly than those that reflect the biliary system. However, there was no complete recovery of impaired hepatic and pancreatic functions throughout the follow-up. PMID- 12886585 TI - [Influence of the protozoa Blastocystis hominis on hepatic function]. PMID- 12886586 TI - [Prevention and treatment of postoperative complications of hepatic echinococcosis]. AB - The paper presents the outcomes of surgical treatment in 117 patients with complicated liver echinococciasis in the past 9 years. Hepatic function and immunity, their pre- and postoperative changes in the combined use of essentiale and T-activin were studied. It has been found that the functional status of the liver, its detoxifying function in particular, largely impairs and immunosuppression develops in patients with complicated liver echinococciasis. The postoperative combined use of essentiale and T-activin led to the normalization of hepatic function and immunity. The combined treatment reduced the incidence of postoperative complications from 34.83 to 17.2%, such as acute hepatic failure, suppuration of a wound and a residual cavity, isolated abdominal abscesses, pleurisy, and pneumonia. PMID- 12886587 TI - [Efficiency of the immunoreflex diagnosis in the assessment of immunological parameters in intestinal parasitic disease]. PMID- 12886588 TI - [Acanthamoeba isolated from child thymus: Acanthamoeba growth during cocultivation with human and murine eukaryotic cells and induction of cytopathogenic effect on these cells]. AB - A cell line of the ameba assigned to the genus Acanthamoeba by morphological and cariotypic signs and by the specific features of its life cycle was isolated from the thymus of a child operated on for heart disease. The line received the name CDHT (Cells Derived from Human Thymus). Actively proliferating mammalian cells maintain the multiplication of Acanthamoeba cells in vitro. Using thymocytes as an example, it was shown that death of these cells occurred through the mechanism of apoptosis. The human thymocytes preincubated with the supernatant obtained through the cocultivation of CDHT and thymocytes may also undergo apoptosis. PMID- 12886589 TI - [Echinococcus granulosus genotyping by PCR-RFLP]. AB - A method to assess the length polymorphism of restriction fragments of polymerase chain reaction (PCR-LPRF) has been developed and used for the genomic typing of E. granulosus. A scheme optimal under our conditions has been chosen for analysis. To test the chosen scheme, 10 E. granulosus samples from our collection made in Russia and Kazakhstan were examined. The material was obtained from 5 sheep, 1 dog, 2 cows, and 1 man. The E. granulosus genotypes detected in Russia and Kazakhstan do not differ from those described earlier. PMID- 12886590 TI - [Developing of new anthelmintics. Evaluation of fluzamide on the new experimental models of intestinal cestode infections]. AB - The efficacy of the original drug fluzamide (a N,S-containing heterocycle derivative) was evaluated by primary screening on a modified model of monoinvasion with the luminal form of Echinococcus multilocularis and on an original model of intestinal mixed invasion of E. multilocularis and Hymenolepsis nana at the immature and mature stages of the parasites in golden hamsters when an experimental host was immunosuppressed with hydrocortisone. The efficacy of fluzamide in an oral dose of 0.3 g/kg against young E. multilocularis and H. nana (the duration of their invasion was 7 days) was 99.8-100%; that against adult cestodes of both species (their maximum age was 28 days) was 100%. The original method of simulation of intestinal cestodiases in laboratory rodents, which is based on the artificial transplantation of cestodes at the prepatent stage of development from the intestine of infected donors to the intestine of healthy recipients through the latter' stomach during drug immunosuppression, showed its high reliability. There is evidence that transplanted immature strobilar cestodes can survive in the stomach, get acclimated, and continue to develop in the recipients' small bowel up to the stage of formation of infective eggs. PMID- 12886591 TI - [Morphofunctional changes in the digestive system of the nematode Passalurus ambiguus (Rudolphi, 1819) after use of the anthelmintics albendazole, fenbendazole, and ivermectin]. AB - The paper presents the results of histological and electron microscopic studies of the tissues of the digestive system of the nematode Passalurus ambiguus (Rudolphi, 1819) after use of the anthelminthics albendazole, fenbendazole, and ivermectine. They demonstrate that albendazole and fenbendazole cause irreversible structural changes. Less pronounced destruction of different parts of the digestive system occurs after the use of ivermectine. All the drugs affect the mid-gut of Passalurus. PMID- 12886592 TI - [Drug resistant tropical malaria in the Republic of Guinea (West Africa)]. PMID- 12886593 TI - [Improvement of a complex of sanitary and health-promoting measures in enterobiasis for children of pediatric institutions and schools]. AB - The use of an improved complex of sanitary and health-promoting measures for children from 2 schools revealed that their infestation with pinworms among first eleventh-form pupils from a boarding school was 42.6%, the above measures covering simultaneously them all with their subsequent repetition every 6 months allowed all the children to be free from pinworms. At school 8, the random use of these measures could reduce pinworm infestation to 4% of the children covered with health-promoting measures. PMID- 12886594 TI - [Methodology for evaluation of disinfectants on the basis of system analysis]. AB - The paper proposes a methodology based on the operation study theory, which can compare disinfectants by a combination of all their properties, which is integrated in the cost of tackling a disinfectological task rather than by the presence or magnitude of these or those useful properties. PMID- 12886595 TI - [Natural focal infections transmitted by Ixodes ticks in the Iaroslavl region: epidemiological aspects]. PMID- 12886596 TI - [Ecology and epidemiology of tick-borne borreliosis in the Ul'ianovsk region]. AB - The parameters that characterize the affection and spread of ticks in the Ulyanovsk Region are presented. A risk for Ixodes tick-borne borreliosis among the urban population is shown. PMID- 12886597 TI - [Incidence of malaria among seamen in Ukraine in 1997-2000]. PMID- 12886598 TI - [Environmental monitoring in the foci of parasitic diseases in the town of Belgorod-Dnestrovsk, Odessa region]. AB - The paper shows a role of environmental objects in the formation of parasitic diseases in Belgorod-Dnestrovsk, developmental changes in intestinal parasitic cenoses in children from closed institutions. To attract parasitologists to the diagnosis of acute intestinal infections of unspecified etiology increases the percent (49.5 to 82.0) of positive results. The findings show what environmental objects should be monitored in the sanitary and parasitological contexts: soil, vegetables, greens in helminthiasis; utensils in preschool children' establishments in contact helminthiasis; drinking water, vegetables, greens in giardiasis. PMID- 12886599 TI - [Effect of cultivation temperature on the extrachromosomal hereditary factors of the causative agent of melioidosis]. AB - The cultivation temperature of Burkholderia pseudomallei has been shown to determine both the direction of morphological dissociation and the prophage induction rate. Inheriting plasmid replicons was found to depend on the temperature conditions during the growth of these bacteria. No influence of B. pseudomallei plasmids pPM1 and pCM2 on the lysogenic state of the bacterial cell and the formation of different B. pseudomallei variants was noted. PMID- 12886600 TI - [Isolation and identification of growth stimulators of Escherichia coli M-17]. AB - The stimulating activity of E. coli M-17 culture fluid (CF) is determined by the action of low-molecular exometabolites, readily soluble in water. The high molecular fraction was removed from CF by ultrafiltration, the growth stimulators were adsorbed on anion exchange resin and eluated at pH 3.1. In further purification HPLC and chromatography on TSK gel HW-40 were used. The identification of compounds was carried out by the methods of thin-layer chromatography, amino acid analysis and 1H-NMR spectroscopy. Glutamic acid (glutamine) and succinic acid proved to be the most active growth autostimulators. The data of biological testing made it possible to believe that CF also contained less active stimulators and/or synergic substances which had no their own activity, but stimulated growth, acting jointly with other compounds. In view of the definite specificity of action observed in initial CF, some differences in the spectrum of growth stimulators of other E. coli strains may be supposed. PMID- 12886601 TI - [Dynamics of the isolation and biological properties of Vibrio cholerae cultures, serogroups 01 and 0139, isolated from water reservoirs and sewage water on the territory of the city of Rostov-on-Don]. AB - The dynamics of the isolation of V. cholerae cultures from various water objects on the territory of Rostov-on-Don during the period of 1994-2001 was analyzed and biological properties of 14 such cultures were studied. In the absence of epidemic complications during the above-mentioned period, a growth in the amount of V. cholerae isolates, serogroups 01 and 0139, including toxigenic V. cholerae 01, was registered. The microbiological and epidemiological aspects of the monitoring of surface reservoirs and sewage were considered and the expediency of the profound and systematic study of its results for epidemiological surveillance on cholera was emphasized. PMID- 12886602 TI - [Morbidity in vaccine-preventable infections in the Russian Federation and measures aimed at their decrease and eradication]. AB - Data on the morbidity in vaccine-preventable infections over the last 10 years in Russia, the analysis of immunization coverage and the level of post-vacination immunity are presented. The conclusions on the epidemic situation with each analyzed infection and decisions on the prospects for their decrease or eradication by the year 2010 are made. PMID- 12886603 TI - [Ixodes tick-borne borreliosis and tick-borne encephalitis in the Udmurt republic: retrospective analysis of their spread]. AB - The primary clinical-anamnestic and laboratory data on 1,062 cases, registered in Udmurtia in 1965-1968 and caused by the bites of taiga ticks (ixodes persulcatus), were retrospectively analyzed. The study revealed that not less than 27% of these cases could be regarded as cases of Ixodes tick-borne borreliosis (ITBB); formerly they were classified with tick-born encephalitis (TBE) or diseases of unclear etiology. The spread of ITBB on the territory of the Udmurt Republic was shown to be similar to that of TBE. PMID- 12886604 TI - [Epidemiology of hemorrhagic fever with renal syndrome in the north Volga-Viatka region]. AB - The results of the ecological and epidemiological study of hemorrhagic fever with the renal syndrome (HFRS) in the Kirov region for the period of 1987-2000 are presented. HFRS morbidity rate varied from 1.3 to 13.7 per 100,000 population, which corresponded to the figures registered in the Volga Vyatka region. Cases of HRFS were mainly (90%) registered in three southern districts of the region. The moderate degree of correlation between morbidity rate and the number of wild mammals (r = 0.55) was established. Periodic morbidity rise occurred every 3 years. In 1998-2000 the contamination of wild mammals with HFRS virus increased and reached 7.6-9.5%. Infection factors connected with everyday human activities somewhat prevailed, industrial and agricultural types of infection constituting 43.8%. Males aged 20-49 years prevailed among the patients. Morbidity rate among urban dwellers exceeded that among town residents. PMID- 12886605 TI - [Efficacy of pneumococcal vaccine in military units]. AB - Pneumococcal vaccine Pneumo-23, used for specific prophylaxis of pneumonia and other pneumococcal infections, was tested in military training units of the North Western, Central and Far Eastern Military Districts. The vaccine used for immunization of servicemen, was shown to have high immunogenicity with no adverse reactions. In the training group of the North Western Military District the epidemiological effectiveness of the vaccine was particularly high a month after immunization and amounted to 83.7%. During the period between month 2 and month 5 after immunization pneumonia morbidity among the immunized servicemen was 6.12 times lower than among the non-immunized ones. In the training units of the Central and Far Eastern Military Districts, where the period of the formation of postvaccinal immunity coincided with the peak of the outbreak of pneumonia, the protective properties of the used batches of the vaccine could be observed as early as during the first month after immunization, which made it possible to recommend this vaccine for urgent prophylaxis in the foci of pneumococcal infection. During the period of 5 months the effectiveness of the vaccine with respect to pneumonia was 62.1-66.2% for all three districts. The effectiveness of the combined immunization of conscripts with vaccines Pneumo-23 and Vaxigrip with respect to pneumonia was higher (78.54%) and the index of effectiveness (4.66) was 1.58 fold greater than in monoimmunization (2.95). The epidemiological effectiveness of the pneumococcal vaccine was high also with respect to other pneumococcal infections: acute bronchitis, acute respiratory diseases of pneumococcal etiology, cases of acute sinusitis and acute otitis. The use of the vaccine for the immunization of servicemen yielded the economic effect equal to 92 US dollars per person. PMID- 12886606 TI - [Experience with vaccine prophylaxis of hepatitis B among different groups of children]. AB - The immunological activity of different vaccines against hepatitis B was evaluated in the Hepatological Center organized on the basis of the Infectious Hospital in Tula. Newborn infants were immunized with the use of the following vaccines: Engerix B (Belgium), Combiotech (Russia), Euvax B (Aventis Pasteur, South Korea). Altogether, after the full course of immunization anti-HBs were detected in 76 children out of 81 (in 93.8%). Vaccine Engerix B, when introduced according to the schedule 0-1-2-12 months, exhibited high immunogenic properties in a group of infants born of women with persistent HBs-antigenemia. Anti-HBs at a concentration exceeding 1000 I.U./I could be detected in 84.6%. In another group of children immunized according to the schedule 0-1-6 months first with vaccine Combiotech at the age of 0 and 1 month, then (at the age of 6 months) with vaccine Euvax, the presence of postvaccinal anti HBs at protective concentration was registered in all children. After immunization against hepatitis B with the use of all above-mentioned vaccines introduced according to both schedules high immunological activity and safety of immunization were noted. PMID- 12886607 TI - [Fc receptors of endothelial cells of cardiac valves: comparison of IgG Fc binding activity of these receptors and of Fc receptors of group A streptococci]. AB - Normal human and rabbit sera, as well as IgG isolated from them, have proved to be capable of reacting with the cells of the valve endothelium of the human and bovine heart. As shown in this study, these reactions are linked with the presence of Fc receptors on the epithelial cells. This is confirmed by the positive reactions of the endothelial cells with the Fc fragments of IgG, as well as with pure antibodies to egg albumin and to group A streptococcal polysaccharide and their complexes. As revealed in this study, Fc receptors on endothelial cells and staphylococcal Fc receptors bind with the definite fraction of normal human serum IgG with, probably, more pronounced cytophil properties. This fraction is not linked with IgG subclasses. The suggestion may be made that the presence of IgG Fc binding activity in group A streptococci, coinciding with the binding activity of Fc receptors in some cells of the human body, is probably of importance for pathogenic streptococci, facilitating their successful invasion. PMID- 12886608 TI - [Rapid laboratory detection of antigens of infective agents of infections and technical means for their realization]. AB - A system of new accelerated and rapid methods for the detection of the antigens of the infective agents of plague, cholera, tularemia and brucellosis were developed on the basis of solid phase immunosuspension tests: the passive hemagglutination (PHA) test and the latex agglutination (LA) test. The immunological and physico-chemical properties of suspensions in the PHA and LA tests made it possible to use extraneous sources of energy (centrifugal acceleration and the electric field) to accelerate these tests. The results of the PHA and LA tests were registered with the use of a densitometer, model Ultrascan 2202, and a tester, model C 34014.2. To apply centrifugal acceleration and the electric field, a laboratory centrifuge and an electrophoretic microchamber were designed. Densitometry was carried out on modified plates and conductometry, with the use of modified electrodes. The time of obtaining the results of the PHA and LA tests was 15-30 minutes with the use of centrifugation and 2-5 minutes in the electric field, which made it possible to regard these tests as rapid. PMID- 12886609 TI - [Detection of hepatitis B virus surface antigen with the use of an optical biosensor]. AB - The detection of hepatitis B virus surface antigen (HBsAg) with the use of a model IAsys+ two-channel optical biosensor is based on the registration of interaction between anti-HBs monoclonal antibodies forming the surface layer of the biochip of the biosensor cuvette and blood serum HBsAg. For the first time a two-channel optical biosensor has been used for the detection of HBsAg in blood serum samples. The comparative analysis of the detection of HBsAg by two methods, viz. with the use of an optical biosensor and the enzyme immunoassay, has demonstrated lower sensitivity, but higher specificity of the detection of this antigen by means of a model IAsys+ biosensor with the biochip, prepared in the process of the work. The main advantages of the biosensor detection lie in the registration of interaction in real time without introducing special markers into the molecules under study. PMID- 12886610 TI - [Clinical and microbiological features of acute enteric mixed infection caused by Hafnia alvei and rotavirus]. AB - The data obtained in the clinical and laboratory study of 72 hospitalized patients with acute enteric infection are presented. The observed outbreak was caused by H. alvei producing heat-stable enterotoxin. The role of this etiological agent is also confirmed by simultaneous occurrence of the disease after using the same foodstuff, a short incubation period, the severity of the course of the disease with pronounced symptoms of neurotoxicosis, a high detection rate of H. alvei in material taken from patients at the acute period of the disease, rapid disappearance of this agent in the period of convalescence and a pronounced rise in the titer of specific antibodies to H. alvei in the dynamics of the disease. At the same time in the feces of 8 patients rotavirus antigen was detected, which, in combination with residual catarrhal phenomena, hyperemia and granularity of the pharynx, yellow stool, was indicative of the simultaneous circulation of rotavirus among these patients. PMID- 12886611 TI - [Microbiocenosis of the mucous membranes of the upper respiratory tract and its correction with acipol in the treatment of patients with acute respiratory virus infections]. AB - Clinical and laboratory evaluation of the preparation Acipol in 45 children greatly susceptible to acute respiratory virus infections (ARVI) was carried out. Simultaneously with the clinical analysis, the study of the microflora of the mucous membranes of the upper respiratory tracts and intestine, as well as the levels of secretory IgA in saliva and coprofiltrates before and after treatment, was made. The inclusion of Acipol into the therapy of ARVI patients facilitated their clinical convalescence, uncomplicated course of the disease, correction of disturbances in the biocenosis of the upper respiratory tracts and increased infectious resistance. PMID- 12886612 TI - [Some specific features of vaginal microflora depending on ecological factors]. AB - In 75 female residents of Vladikavkaz, aged 18-45 years, who had applied to the gynecological department with complaints of vaginal discharge, the qualitative and quantitative composition of vaginal microflora was studied. All these women were divided into 2 groups: group 1 including 38 women living in ecologically unfavorable districts of the city and group 2 including 37 women living under ecologically favorable conditions. The relationship between the ecological situation of the districts of residence and vaginal microflora in women of the reproductive age was established. Unfavorable exogenous factors were found to lead to the development of vaginal dysbacteriosis: a sharp decrease in the amount of lactoflora or its complete absence accompanied by increased amount of staphylococci, enterococci, Escherichia coli, Proteus, Klebsiella and yeast-like fungi of the genus Candida. PMID- 12886613 TI - [Dynamics of antibiotic resistance of Staphylococcus aureus isolated from chronic osteomyelitis patients]. AB - The level of antibiotic resistance of 365 S.aureus strains, isolated from chronic osteomyelitis patients in 1990 and 2000, was determined. As revealed in this study, over 10 years the isolation rate of strains adapted to the antimicrobial action of all tested preparations increased in some degree. The increase of the resistance level to aminoglycosides was most significant. The proportion of polyresistant strains was 9.1% in 1990 and 30.5% in 2000. The isolation rate of methicillin-resistant S. aureus (MRSA) did not increase during these 10 years, but the range of preparations suitable for the treatment of MRSA-infected patients was shown to become essentially narrower. These data confirm the view on the possibility of further decrease in the effectiveness of the conservative treatment of chronic osteomyelitis due to the rapid growth of the antibiotic resistance of S. aureus. PMID- 12886614 TI - [Regional features of sensitization of children to some noninfectious allergens]. AB - The results of epidemiological investigations within the frames of the ISAAC program are presented. Out of 2,870 children, 172 children (6.0%) with sensitization to some noninfectious (insect, pollen, medicinal) allergens were detected. The number of allergic children was found to be 10.4 +/- 1.8 to 28.2 +/ 3.0 per 1000 of urban child population. Specific features of the sensitization of urban children to regional insect, pollen and medicinal allergens were established. The clinical course of allergic diseases in children was characterized by polymorphism and a great variety of clinical symptoms with frequent lesions of the gastrointestinal tract and functional disturbances of the central nervous system, which adversely affected the school studies of children and the quality of their life. In the development of allergy in children such factors of risk as hereditary predisposition to allergy, allergic diathesis, focal infections and artificial feeding during the first year of life were also of great importance. PMID- 12886615 TI - [Evolution of epidemiology]. AB - The emergence, formation and stages of the development of today epidemiology of infectious diseases are analyzed. Differences in views concerning the essence of epidemiology as science between Russian and foreign schools are indicated. The brief description of the main stages in rendering basic concepts of Russian epidemiology more extensive and profound, including the gradual alteration of the view on epidemiology as the science dealing only with the epidemic process, is presented. In this connection new data on the epidemiological specificity of zoonotic, and especially sapronotic infections, leading to an essenmtal correction of the main postulates of epidemiology, are analyzed. Broader notions of the reservoirs of the causative agents of infections are given, the discrete character of the epidemic (epizootic) process, the role of the latent forms of bacteria in their preservation both in the environment and in the human body in the course of the infectious process are postulated. The complexity of the term "emerging infections" is discussed and 3 variants of its interpretation are proposed. The increasing role of anthropogenic factors, including technogenic effects, is emphasized; in future these factors may be capable of accelerating the man-made evolution of epidemiology. PMID- 12886616 TI - [Methods of molecular epidemiology of infections caused by group B streptococci]. AB - In this review the comparative analysis of advantages and disadvantages of the known methods for the immunological diagnostics of streptococcal infections (serotyping, phage typing), as well as the methods of molecular epidemiology (PCR typing with the use of "disseminated" priming, ribotyping, electrophoresis in the pulsing electric field, etc.), is presented. Essentially new approaches, capable of being used in future for the control of epidemically topical clones of group B streptococci, are discussed. PMID- 12886617 TI - [Minor fatty acids of biological fluids of urogenital organs and their significance in the diagnosis of inflammatory processes]. AB - Minor lipid components (less than 1%), not occurring in the cells of mammals, have been detected in the profile of fatty acids (FA) in urine and vaginal fluid in females, as well as in ejaculate in males, by the method of gas chromatography and mass spectrometry. Their origin is supposedly attributed to microorganisms colonizing urogenital organs under normal conditions and in infectious pathology. Minor FA in the biological fluids of the urogenital tract (UGT) in man are compared with the FA composition of pure microbial cultures. The statistical analysis of FA concentrations in 500 specimens of UGT biological fluids indicates that lipid markers of microbial origin (in UGT inflammatory diseases) have a range of quantitative changes exceeding the limits of values characteristic of healthy persons and in a number of cases correlate with the diagnosis of the disease or the results of the bacteriological study. The reliability of the interpretation of the results of the marker analysis is confirmed by the difference in the composition of UGT microorganisms under normal conditions and in pathology, correlation with the clinical picture of pathological deviations, a decrease (to zero or normal values) in the concentration of the markers of pathogens as the result of treatment with antibiotics, as well as by the adequate composition of UGT microorganisms, determined by the marker method in comparison with the cultural biochemical method. PMID- 12886618 TI - [Biological effects of interferon, produced by recombinant bacteria of the probiotic preparation subalin]. AB - The present review deals with the analysis of biological and functional activities of recombinant bacteria Bacillus subtilis IF-alpha 2335 are producing a human interferon. The interferon-producing bacteria are constructed on a basis commercial probiotic strain B.subtilis 2335, carrying a recombinant plasmid pMBM 105 with the gene of human alpha-2 interferon. The implementation of the recombinant strain in the preparation probiotic, received a designation "Subalin", necessitates to verify a number of immunologic activities and to perform successive protective effects. Interferon, synthesized by recombinant bacteria shows the activity on macroorganism at oral and rectal application of preparation. Subalin was shown antivirus and antitumor activity and preservation by recombinat bacteria of antagonistic properties. The mechanisms of the positive effect of subalin were considered: this effect was shown to be due to the action of interferon excreted by recombinant bacteria into the mucous of different biotopes of host. PMID- 12886619 TI - [Discussion on the problems of general epidemiology]. AB - The key problems of general epidemiology: the subject, method, causality in the epidemic process, the main categories and laws, the systematization of infectious pathology and epidemiological terminology are under discussion. It is at this stage when students should form valid notions, well grounded on scientific facts and practical experience, concerning the epidemiology of noninfectious diseases with due regard to the existing differences in the interpretation of the causes of the epidemic process in noninfectious diseases and to the characteristic distribution of noninfectious diseases among the population under the influence of environmental factors without any participation of parasitic systems. The review of the notion apparatus of the theoretical epidemiology and putting it in order in accordance with general biological concepts are proposed in connection with the necessity of differentiation between the epidemic process proper (in cases of anthroponosis) and the epidemic manifestations of epizoonotic and epiphytotic processes (in cases of zoonoses and sapronoses). PMID- 12886620 TI - [Viral hepatitis A: epidemiologic role of parenteral route of transmission]. PMID- 12886621 TI - [Diagnosis, treatment and prevention of Ixodes tick-borne borreliosis in the Ul'ianovsk region]. AB - The level of morbidity in Ixodes tick-borne borreliosis, the analysis of the quality of diagnostics and the results of treatment are presented. The necessity of taking prophylactic measures is substantiated. PMID- 12886622 TI - [Alteration of phagocyte functional activity depending on the presence of pR50 plasmid in bacteria]. AB - The relationship between the presence, or the absence, of conjugative plasmid pR50 detected in Klebsiella oxytoca 89, in the isogenic pairs of attenuated strains Shigella flexneri 2a 516 Near and Salmonella typhimurium 129 Rifr and the regeneration of the active forms of oxygen by mouse peritoneal phagocytizing cells was studied. As indicated by the data obtained in the course of the experiment, plasmids pR50 could influence the chemiluminescent response of phagocytes. The inhibition of the synthesis of oxygen metabolites was found to occur at the primary stage of the infectious process, that later this synthesis increased, that facilitated the survival of the animals infected with the cultures carrying plasmid pR50. PMID- 12886623 TI - [Evaluation of the polymerase chain reaction effectiveness in the investigation of out-breaks of pseudotuberculosis]. AB - Data on the investigation of pseudotuberculosis epidemic outbreaks with the use of the polymerase chain reaction (PCR), are presented. Specific fragments of Yersinia pseudotuberculosis DNA were detected in 81.25% of patients, in 46.83% of cases confirmed by the isolation of Y. pseudotuberculosis cultures. The study of washings from vegetables and equipment in vegetable stores and kitchens yielded positive results in PCR in 8.52% and the survey of rodents--in 3.85% of cases. In the course of the bacteriological study of these specimens 6 Y. pseudotuberculosis cultures were isolated: 3--from vegetable products, 1--from a Norway rat and 2--from house mice. The coincidence of the data obtained by bacteriological study and PCR showed that the latter method gave objective results, while being capable of ensuring rather rapid analysis. PCR should be regarded as a signal test for the bacteriological search of the definite infective agent in the material under study. PMID- 12886624 TI - [Cultivation of microorganisms in micro-volumes of culture media]. AB - Studies on the cultivation of aerobic microorganisms in microdrops of growth medium, immobilized in a layer of highly dispersed disconnector (Aerosil), were carried out. The dispersion of the growth medium and its immobilization in a layer of Aerosil was found to produce no influence on the morphological, cultural and biochemical properties of Serratia marcescens cells, determined both immediately after treatment with the field and some time after storage of the treated suspensions. On the contrary, the duration of the action of ferromagnetic bodies produced essential influence on the viability of the microorganisms in dispersed growth medium and on the accumulation of cells in the course of subsequent cultivation. The development of cells grown in the microdrops of the nutriuent medium was similar to that of cells grown in submerged cultivation, following the S-shaped curve. With the increase of the seed dose a drop in the initial growth rate occurred and the accumulation of microbial cells increased by the end of the process. The greatest increase of biomass was observed with minimal seed doses. With a rise in the concentration of amino nitrogen in the growth medium the yield of cells increased only till this concentration reached 320 mg%. Under these conditions the specific growth rate also increased. The accumulation of cells was greater and occurred earlier when microorganisms were cultivated in immobilized state in powder. PMID- 12886625 TI - [Effect of various amino acids and ammonium salts in a synthetic culture media on the cholera enterotoxin production]. AB - The influence of amino acids and ammonium salts on the production of cholera enterotoxin (CT) by 3 Vibrio cholerae strains of different biovars and serogroups was evaluated. As revealed in this study, toxin formation in each of the strains was quantitatively and qualitatively determined by their individual sets of amino acids. The amino acid compositions ensuring the maximum production of CT by the V. cholerae strains under study were formed. The use of ammonium salts as the only source of nitrogen in the composition of a synthetic nutrient medium for the accumulation of CT was shown to be inexpedient. PMID- 12886626 TI - [HIV infection in Russia and Commonwealth of Independent States: current situation and prospective]. AB - The dynamics of the spread of HIV infection in Russia and the CIS countries is presented. The relationship between the spread of HIV infection and an increase in the number of injection drug users is substantiated. The factors inhibiting the spread of the epidemic of HIV/AIDS in Russia and the CIS countries are analyzed with due regard to the world experience. Attention is paid to the development of the epidemic situation in HIV infection due to, similarly to Russia and to more than 100 countries of the world, drug addiction. The effectiveness of the activity of the Russian health service organs in controlling this epidemic is emphasized. The prospects of prophylactic measures for controlling HIV infection in Russia and the CIS countries are determined. PMID- 12886627 TI - [Postvaccinal immunity and immunological aspects of Haemophilus influenzae carrier state in children of different age groups after the administration of "Act-HIB" vaccine]. AB - The article deals with H. influenzae (different serotypes) carrier state and immune response before and after the administration of the vaccine "Act-HIB" to children of different age groups. Children aged up to 1 year and over 1 year have been found to differ in the dynamics of carrier state and in the concentration of antibodies of different classes to the antigens of this infective agent, which makes it necessary to carry out their early immunization with a view to ensure their protection from H. influenzae infection. PMID- 12886628 TI - [Morphological alterations in animal organs after the injection of acellular pertussis vaccine with immunomodulator]. AB - Toxic properties of acellular pertussis vaccine (APV) and morphological changes in white mice in response to intramuscular injection of APV (without or with immunomodulator glucosaminylmuramyl dipeptide-GMDP) were under study. APV used in these experiments was developed at the Mechnikov Research Institute for Vaccines and Sera (the Russian Acad. Med. Sci.) on the basis of Bordetella pertussis cultures in synthetic fluid culture media. In experiments on acute and chronic toxicity of APV (without GMDP) increased tissue immunity reactions in spleen, thymus, liver, lungs and intestinal wall was detected. There was no difference in immunomorphological reactions in mice receiving APV with different doses of GMDP, but some difference was observed in time dynamics of tissue immunity reactions. A small dose of GMDP should be preferred (0.0001 microgram) which results in gradual growth of tissue immunity reactions less pronounced toxic reactions caused be the APV injection. PMID- 12886629 TI - [Dynamics of some characteristics of humoral immunity, vaginal interferon status in pregnant women with the relapse of chronic pyelonephritis]. AB - 65 pregnant women with the exacerbation of chronic pyelonephritis in the III trimester of gestation and 34 healthy pregnant women were examined. The quantitative content of immunoglobulins, the activity of interferon in cervico = vaginal washings and the composition of the vaginal microflora were determined. All patients with the relapse of chronic pyelonephritis exhibited disturbances in the normal microbiocenosis of the genitals and the dysfunction of the local immunity of the genital system, accompanied with a decrease in serum and secretory IgA, an increase in the amount of IgG and IgM, increased interferon activity. Pregnant women with the relapse of chronic pyelonephritis received, in addition to traditional therapy, local treatment with Kipferon suppositories, an immunomodulating preparation. The study revealed that the use of this preparation normalized the characteristics of local immunity, the composition of the microflora' of the genitals and led to the disappearance of the clinical symptoms of the disease. PMID- 12886630 TI - [The possibility of the detection of one more tick-borne infection--babesiosis- on the territory of Russia]. AB - A total of 739 taiga ticks of ixodes persulcatus species, obtained in the recreational zone of St. Petersburg, were studied for the presence of Babesia sp. with polymerase chain reaction. All these ticks underwent the preliminary examination for the presence of Borrelia (3 species), Ehrlichia (2 species) and tick-borne encephalitis (TBE) virus. In 7 cases Babesia were detected among 413 ticks containing other pathogens. Among 326 ticks no Babesia were detected, as well as no other pathogens. All ticks having Babesia were also found to contain Borrelia species: B. afzelii, B. garinii, or both (1 case). In one female tick, in addition to Babesia, also B. garinii and TBE virus were determined. The data thus obtained should draw special attention not only of parasitologists, epidemiologists and microbiologists studying ticks obtained from natural sources, but also of clinicists who should consider the possibility of mixed infection, when one infection may mask the presence of some other infection, in particular babesiosis. Due to rare occurrence of Babesia in ticks and the presence of mixed infection difficulties may arise in the detection of Babesia. PMID- 12886631 TI - [Detection of Treponema pallidum DNA and RNA in clinical material from patients with syphilis at different stages]. AB - The results of the development and approval of methods for the detection of T. pallidum DNA and 16S rRNA in clinical material (blood plasma, serous exudates) are presented. T. pallidum DNA was detected with the use of primers to the gene coding protein with a moleculat weight of 47 kD and T. pallidum RNA, with the primers to gene 16S rRNA. The isolation, reverse transcription and amplification of DNA and RNA was carried out in the presence of inner DNA and RNA control respectively. The analytical sensitivity of the developed method was 400 DNA copies per ml. The characteristics of analytical and diagnostic specificity were 100%. The specimens of blood plasma, taken from 292 patients with syphilis at different stages before specific antibacterial therapy, were tested by the PCR. The detection rate of T. pallidum DNA and RNA in blood plasma was, respectively, 91% and 100% in primary seropositive syphilis, 68% and 79% in secondary early syphilis, 19% and 26% in latent unverified syphilis. In secondary relapsing syphilis T. pallidum DNA and RNA were detected in 92% and in latent early syphilis, in 14% of patients. T. pallidum nucleic acids were detected in 1 patient at the seronegative period of primary syphilis. No positive result was obtained in the PCR analysis in any of the patients with diagnosed seroresistance, latent late syphilis and tertiary syphilis. In the study of material taken from chancres of 11 syphilis patients the data obtained by dark field microscopy and the PCR analysis completely coincided. PMID- 12886632 TI - [Comparative information value of humoral immunity characteristics in some bacterial and viral infections]. AB - Serological examination of 144 patients with different bacterial and viral infections was carried out. Antibodies to Brucella were detected in blood serum in 42 patients (85.7%) with the average titer of 1:996 and in saliva in 41 patients (83.7%) with the average titer of 1:567 by passive hemagglutination test with brucella erythrocyte diagnosticum. Out of 26 dysentery patients, antibodies in blood serum were detected in the diagnostic titer in 17 patients (65.4%) with the average titer of 1:282 and in saliva in 21 patients (80.8%) in the titer of 1:100 and higher. Anti-HAV and anti-HBc IgM were detected in specimens of saliva from patients with serologically confirmed viral hepatitis A and B in 100% of cases. The presence of HBsAg in saliva from hepatitis B patients was established in 95.4% of cases. In blood serum and in specimens of saliva anti-HCV IgM were detected in 100% and 85.7% of cases respectively. Out of 25 women with aggravated obstetric history, IgG antibodies to CMV were detected in blood serum in 23 women (88.5%) and in saliva in 22 women (84.6%). The results of these investigations revealed that the detection rate of antibodies in blood serum and saliva in cases of infections, both bacterial (brucellosis, shigellosis) and viral (hepatitis A, B, C and CMV infection), was not essentially different. The simplicity of obtaining material for analysis make it possible to recommend the use of saliva for diagnosing bacterial and viral infections, especially in mass epidemiological surveys. PMID- 12886633 TI - [Generation of antibodies to Yersinia pseudotuberculosis antigens using the colloid gold particles as an adjuvant]. AB - Serum titer achieved while producing antibodies to Y. pseudotuberculosis surface antigens with the use of colloid gold as adjuvant was as high as that achieved with the use of Freund's complete adjuvant (1:10,240). Still the amount of the antigen introduced when colloid gold particles were used as adjuvant was lower by 2 orders. The study revealed that colloid gold used as antigen carrier activated the phagocytic activity of lymphoid cells. PMID- 12886634 TI - [Evaluation on monkeys of reactogenicity and effectiveness of the complex immunoglobulin preparation formulation]. AB - Experiments on 75 monkeys, 20--healthy and 75--with diarrhea, showed that ready medicinal forms of the complex immunoglobulin preparation in tablets, capsules or in a dried form (vials) were safe, well tolerated, clinically and bacteriologically effective. When used simultaneously with antiparasitic treatment, this preparation, irrespective of the kind of its ready medicinal form, showed high curative effect (80-90%). The diarrhea ceasing was accompanied by the gradual normalization of intestinal microbiocenosis (an increased concentrations of Escherichia coli and Bifidobacterium, a decreased amount of opportunistic bacteria). The complex method of the treatment of diarrhea in monkeys (antibiotics, trichopol, phage and probiotics) made it possible to achieve curative effect only in 60% of cases, which required the additional course of treatment, namely the use of rehydration therapy. PMID- 12886635 TI - [Microflora of the large intestine in patients with connective tissue dysplasia]. AB - The specific features of intestinal aerobic and anaerobic intestinal microflora in children with nondifferentiated connective tissue dysplasia were under study. A high rate of dysbiotic disturbances was noted in patients with connective tissue dysplasia in comparison with healthy persons. Profound quantitative and qualitative changes in the biocenosis of the intestine were detected in patients with the pathology of the gastrointestinal tract as well as that of the locomotor system. Changes in the species composition and the persistence properties of the intestinal microflora may serve as pathogenetic factors in the development of connective tissue dysplasia. PMID- 12886636 TI - [Microflora of the nasal mucosa in allergic perennial and infectious rhinitis]. AB - The microbiological study of 69 patients with allergic annual rhinitis (AAR) and infectious rhinitis (IR) was carried out. In AAR the isolated representatives of 15 genera and 40 species were distributed in 2 to 7 component; in IR the isolated representatives of 16 genera and 25 species were grouped in 2 to 4-component associations. In AAR Staphylococcus aureus was found to belong to the main species and in IR, S. aureus and S. epidermidis, while the number of species regarded as occasional in AAR was 7 (S. auricularis, S. cohnii, S. hominis, S. haemolyticus, S. warneri, S. apitis, S. schleiferi). Differences in the distribution of Neisseria, nonfermenting Gram negative bacteria, Streptococcus in associations in cases of AAR and IR were established. In AAR Corynebacterium pseudodiphthericum and in IR C. pseudotuberculosis were the dominant species. PMID- 12886637 TI - [Bacterial infectious process in patients with chronic obliterating bronchiolitis]. AB - Chronic obliterating bronchiolitis in children is characterized by the presence of chronic multifactor inflammatory infectious process with Streptococcus pneumoniae and Haemophilus influenzae being the leading causative agents. The inflammatory process is induced and maintained by S. pneumoniae and H. influenzae or by the association of these two pathogens, but the severity of the clinical course of obliterating bronchiolitis is determined by H. influenzae, which is confirmed by its high detection rate at the phase of exacerbation in patients with pronounced bronchial obstruction. PMID- 12886638 TI - [Dependence of the expression of the biological features of Vibrio cholerae on the conditions of their cultivation]. AB - The biological activity of toxigenic and non-toxigenic V. cholerae supernatants was found to depend on the cultivation medium. The use of iron-free tryptone medium made it possible to obtain supernatants of toxigenic V. cholerae with haemolytic activity and destructive action on passaged cell cultures. In the experimental infection of suckling rabbits the influence of the cultivation conditions of V. cholerae on the character and expression of their pathogenic properties was determined. The dissemination of V. cholerae into the internal organs of rabbits after their infection with both toxigenic and non-toxigenic strains correlated neither with the cultivation conditions of these strains, nor with the character of changes in the intestine of the infected animals. PMID- 12886639 TI - [Detection of lymphocytes binding erythrocytes conjugated with Salmonella antigens]. AB - Immune reagents for the detection of specific antigen-binding lymphocytes (ABL) with respect to different Salmonells antigens were developed. Rabbits were immunized with killed S. typhi and other salmonellae containing cross-reacting antigens, and the dynamics of the formation of ASL of each specificity was studied. Differences in the time of the appearance of ASL with receptors to thymus-independent (09, 12 or Vi) and thymus-dependent (Hd) antigens were studied. The relative content of ASL, determined with the use of immune reagents prepared from S. typhi antigens, was higher, on the whole, in rabbits immunized with S. typhi than in rabbits immunized with salmonellae containing one of cross reacting antigens (S. enteritidis--09, 12; S. paratyphi C--Vi; S. virginia--Hd). PMID- 12886640 TI - [Antigenic specificity of Vibrio cholerae O139 nitrosoguanidine mutants]. AB - The action of nitrosoguanidine (NG) on the culture of V. cholerae O139 P-16064 resulted in the appearance of mutant 16064 NG6, not agglutinating with commercial diagnostic serum O139. Its incapacity of agglutination was due to the sorption of the specific serum with strains V. cholerae O22 and R-variant RCA-385, which caused the loss of antibodies to common determinants. Experiments with the sorption of immune sera made it possible to suggest that one of the determinants of LPS O139, phosphate-galactose, was absent in NG mutant. PMID- 12886641 TI - [Epidemiological and economic effectiveness of immunization of adult and children against influenza and acute respiratory viral infections with the vaccine Grippol]. AB - Materials on the characterization of the vaccine Grippol, indications for its use and the results of mass use are presented. Analysis of the morbidity level in influenza and acute respiratory viral infections (ARVI) among the vaccinated persons are indicative of considerably decreased level. The coefficient of the epidemiological effectiveness of Grippol in the immunization of children has proved to be 75-95%. The conclusion has been made that the vaccine Grippol is sufficiently effective and safe for use by Russian public health service. PMID- 12886642 TI - [Development of HIV infection epidemic among intravenous drug users in the Sverdlovsky region]. AB - The Sverdlovsky region takes the fourth place among the 89 subjects of the Russian Federation by the number of registered cases of HIV infection. Drug addiction has reached an excessive scale in this region: according to the data of express evaluation carried out by the Regional Narcological Hospital and the Regional AIDS Center, not less than 7-8% of persons aged 15-30 years constantly use injection drugs. The large-scale epidemic of injection drug addiction has led to the rapid spread of HIV among addicts. The first penetration of HIV into this population took place in 1996 and, starting from the year 2000, rapid, development of the epidemic began, taking the character of an avalanche. The peak of new cases of HIV infection fell on 2001 (9,230 cases were registered). The concentrated stage of the epidemic development is observed in the region at present, the prevalence of HIV infection among drug addicts being 13.8%. Children born from HIV-infected drug addicted mothers now represent a new risk group due to great difficulties in the prophylaxis of the vertical virus transmission. PMID- 12886643 TI - [Epidemiological surveillance on natural-borne infections in the Ulyanovsk region]. AB - The analysis of natural foci of haemorrhagic fever with renal syndrome (HFRS) and tick-borne borreliosis on the territory of the Ulyanovsk region is presented. Reduced rat control interventions result in a wide spread of these and other infections with natural focality. Monitoring of natural foci of HFRS, tick-borne borreliosis and arbovirus infections, their typing and determination of main reservoirs are the most promising among epidemiological surveillance methods. PMID- 12886644 TI - [Characterization of highly purified metalloprotease produced by Bacillus subtilis]. AB - Proteolytic enzyme produced by Bacillus subtilis is characterized as typical metalloprotease with a molecular weight of 27.9 kD; the enzyme shows its highest activity at pH 7.0-9.0, possesses substrate specificity with respect to different proteins, its temperature optimum is 52 degrees C and its specific activity exceeds that of all known commercial analogues. At 37 degrees C the enzyme is half inactivated in 72 hours. PMID- 12886645 TI - [Inhibitors of the polymerase chain reaction]. AB - The information on the applied aspects of the polymerase chain reaction (PCR) is updated. In particular, the main inhibitor of PCR, considerably decreasing the sensitivity of the method both at the lysis stage of the tested material and due to the degradation of the DNA matrix and primers and/or to the direct inhibition of the activity of DNA polymerase, are described. The compounds, most frequently distorting the course of the reaction while testing clinical blood samples, bioptic samples, sputum, etc., are characterized. Testing concrete clinical material with the use of PCR was shown to require differentiated approach both at the stage of choosing the adequate method for the preparation of samples and at all other stages, including, e.g., the corresponding DNA polymerases or at the stage of heating for decreasing endonuclease activity or for IgG denaturation. Information on the causes of false negative results of PCR and the variants of their elimination, useful under practical laboratory conditions, is given. PMID- 12886646 TI - [Molecular epidemiology of infections caused by methicillin-resistant staphylococci]. AB - The review deals with the periodicity of the spread of methicillin-resistant S. aureus (MRSA) strains during the last 40 years, the mechanism of their resistance to methicillin and other beta-lactamic antibiotics, the genetic control of methicillin resistance, the genome organization of mec DNA and its possible cause, as well as the organization of epidemiological surveillance on MSRA in hospitals. The problem of changes in the epidemiology of staphylococcal infections due to the appearance of MRSA in the absence of contacts with carriers, treatment with antibiotics or stay in a hospital is discussed. The concern of public health authorities in connection with the emergence of MRSA strains, moderately resistant or resistant to vancomycin, is also discussed. The most promising programs of the MRSA study, as well as the optimum programs introduced in economically developed counties for the control of hospital infections caused by MRSA, are considered. PMID- 12886647 TI - [Bacteriocins: criteria, classification, characteristics, methods of detection]. AB - The review on bacteriocins of Gram negative and Gram positive bacteria. Criteria making it possible to regard antagonistic substances as bateriocins or bacteriocin-like substances and on their classification are presented. Examples of bacteriocins naming depending on the taxonomic position of the producer culture are given. Information on the physico-chemical and biological properties of bacteriocins and their purification is presented as well as on detection tools of bacteriocins in microorganisms and evaluation of the producer activity of the bacteriological culture. PMID- 12886648 TI - [Spore probiotics]. AB - Data on probiotic preparations prepared from spore-forming bacteria is reviewed. Information on indications for use, effectiveness and the mechanism of action is given. Preparations being produced in Russia (Biosporin, Sporobacterin and Bactosporin) are evaluated in detail as well as the prospects of the application of probiotics from spore-forming bacteria. PMID- 12886649 TI - [Prospects and problems of current disinfectology]. AB - Topical problems of modern disinfectology are formulated in connection with differences in the resistance of different microbial groups, species, strains to disinfectants, as well as with the "critical ability" of instruments and, accordingly, with different requirements to the necessary levels of decontamination. Factors determining the effectivity of disinfection are described. Requirements to "ideal" disinfectants and disinfectological technologies are considered. The problems of harmonization of approaches to the evaluation of Russian and foreign disinfectological methods and bringing them closer together are discussed. PMID- 12886650 TI - Beyond the walls. AB - For more than 100 years, Michigan's nonprofit community hospitals have provided high-quality health care to all who come to their doors, regardless of the individual's ability to pay. This mission of compassionate caring is a long standing tradition and one that defines what hospitals are all about. Michigan's nonprofit community hospitals serve as the health care safety net for the uninsured and underinsured. Their doors are open 24 hours a day, seven days a week, 365 days a year. Michigan citizens have come to expect nothing less and oftentimes more. But providing high-quality and accessible acute care is only part of what makes Michigan's nonprofit hospitals an essential community asset. PMID- 12886651 TI - Health-related cost-to-benefit measurement. A difficult sell in a "show me the money" era. AB - "Cost to benefit" is both a generic term and a specific technique used to describe and guide budgetary and investment scenarios. The methodology is based on computer mathematical models to prospectively define the short- and long-term financial gain or burden that may be produced by investing today's dollars in tomorrow's programs, technology or facilities. PMID- 12886652 TI - Clearing the final HIPAA hurdle: preparing for compliance with the HIPAA Security Rule. AB - On Feb. 20, while most health care providers were in the midst of last-minute preparations to comply with the Health Insurance Portability and Accountability Act (HIPAA) Privacy Rule, the Department of Health and Human Services (DHHS) published the final HIPAA Security Rule. PMID- 12886653 TI - Building a healthier northern Michigan. AB - Over the last 10 years, hospitals in the north central region of Michigan have collaborated with human service agencies across a 21-country area of northern Lower Michigan on the Community Health Assessment Project: Building A Healthier Northern Michigan. A survey was completed in 1995 that provided insight into the health status of adults in the region. The survey helped communities identify health priorities and implement plans to address them. PMID- 12886654 TI - The isolated leader: extraverted and introverted styles. AB - When an executive moves up the hierarchal ladder in the organization, the promotion is often accompanied by an increasing sense of isolation and loneliness. Goleman, Boyatzis and McKee coined the phrase "CEO disease" to describe the isolation of top executives in their book Primal Leadership. It refers to an information vacuum around leaders, created when people withhold important and sometimes unpleasant information. PMID- 12886655 TI - The cost of being couch potato. AB - The scientific evidence is clear--regular physical activity has powerful positive effects on both physical and psychological health. Conversely, physical inactivity has a high human cost in terms of health. It shortens years of life, decreases quality of life, and limits functional independence. These health effects arise from physical inactivity's contribution to cardiovascular disease, obesity, diabetes, osteoporosis, mental health disorders, and some cancers (i.e., colon and breast). PMID- 12886656 TI - Demystifying "QI". AB - In the continuing pursuit of performance excellence, many quality improvement models such as CQI, TQM, ISO, Six Sigma, and the Malcolm Baldrige criteria for performance excellence have been introduced and implemented with varying degrees of success. PMID- 12886657 TI - Nurse retention: outrunning the bear. AB - Did you hear the one about the hospital CEO who was hiking in the forest with the director of nursing? As they were heading down a trail, suddenly a bear jumped out of a bush and started chasing them. Both hikers started running for their lives when all of a sudden the CEO stopped, sat down on a rock and started to put on running shoes. The director of nursing said, "What are you doing? You can't outrun a bear!" The CEO replied, "I don't have to outrun the bear. I only have to outrun you." PMID- 12886658 TI - From "them" to "us". Going where others have gone before. AB - To Mary Gallagher, Any Town Hospital's new CEO, the growing need for physicians and administrators to work together to solve the challenges before them seemed obvious. Physicians agreed, but little progress had occurred. Recessionary government budgets and employer pushback through reduced benefits and increased co-payments seemed ominous. Medical malpractice premiums were spiraling upwards, pharmaceutical costs were escalating, and the percentage of health care insurance premiums going to medical claims was decreasing. The basis for competition was shifting from managed care contracting to quality, demanding further financial investments (e.g., computer technology). PMID- 12886659 TI - Back to basics: giving attention to surgical infection prevention. AB - When it comes to postoperative infection, there should be no argument about the business case for quality. Postoperative infection is a major cause of patient injury, mortality and health care cost. An estimated 2.6 percent of the nearly 30 million operations each year are complicated by surgical site infections, and patients with infections have twice the incidence of mortality. PMID- 12886660 TI - What to do when trying harder is not the answer. AB - I talk with many health care senior managers and executives. In their most candid moments, they admit to finding it hard to sleep at night. What's keeping them awake? The overwhelming demands of managing 21st century health care. PMID- 12886661 TI - Repairing damaged care. AB - These are times of great change and upheaval in health care. More and more people are uninsured, consumers and employers pay substantially higher health insurance premiums for care, there is a shortage of nurses, and doctors go on strike. And these facts don't take into account that hospitals are going bankrupt while searching for their roles in the 21st century, the population is aging, the growth of alternative and complementary health care, the biotechnology and genetics revolution, the depersonalization of health care, and the huge cutbacks in federal and state government funding of health care and other human services. PMID- 12886662 TI - Healthcare Lean. AB - Lean Thinking is an integrated approach to designing, doing and improving the work of people that have come together to produce and deliver goods, services and information. Healthcare Lean is based on the Toyota production system and applies concepts and techniques of Lean Thinking to hospitals and physician practices. PMID- 12886663 TI - Managing board relationships in tough times. AB - Scandals have dominated newspaper headlines, bringing corporate directors under increasing scrutiny. These issues continue to elicit public outrage about wrongdoing and ultimately resulted in standards for for-profit corporate board members through the enactment of the Sarbanes Oxley Act in 2002. Although all board members must address corporate responsibility issues, directors of health care organizations also have responsibilities unique to the health care industry that must be met in relation to corporate compliance requirements. PMID- 12886664 TI - Breaking through to improved community health. AB - Michigan's nonprofit community hospitals serve the state's citizens in extraordinary ways--regardless of whether anyone pays for that service. In 2001, these community-based organizations provided $885 million in free health care to area residents who were uninsured or otherwise could not pay for it. In addition, the state's nonprofit hospitals contributed more than $276 million in services such as health education, outreach, screening, counseling and free clinics. The total worth of these traditional and nontraditional benefits to Michigan communities in 2001 was a staggering $1.16 billion. PMID- 12886665 TI - Good health: the generic alternative. AB - By the year 2011, medical expenditures in the United States are projected to double from today's outlay of $1.5 trillion. This is in an environment where purchasers of health care coverage already think the rate of growth in health care expenditures is unsustainable. PMID- 12886666 TI - Rethinking nutritional support for persons with cancer cachexia. AB - Cancer cachexia is a poorly understood syndrome of anorexia, weight loss, and muscle wasting that negatively impacts quality of life and survival in cancer patients. Research has clearly implicated pro-inflammatory cytokines in the biology of cancer cachexia. More recent research implicates products of arachidonic acid and suggests that cachexia may be a chronic inflammatory condition rather than a nutritional aberration. To date, nutritional support to slow weight loss has focused primarily on increasing calorie intake. Alternatively, many foods contain factors that can modulate the synthesis or activity of pro-inflammatory mediators, especially the synthesis of prostaglandin E2 from arachidonic acid. These factors and foods are sometimes called nutraceuticals, and research is needed to evaluate their efficacy in combating cancer cachexia. PMID- 12886667 TI - The relationship between chronic pain, immune function, depression, and health behaviors. AB - The objectives of this study were 1) to determine the relationships between pain perceptions, immune function, depression and health behaviors and 2) to examine the effects of chronic pain on immune function using depression and health behaviors as covariates. A nonexperimental cross-sectional design was used to compare 24 chronic back pain patients with 33 asymptomatic no-pain healthy subjects. Immune function measures using natural killer (NK) cell activity, percent and numbers of NK cells, and T-cell lymphocyte proliferation with mitogens concanavalin A (ConA) and pytohemagglutinin (PHA) as well as self-report measures for depression and health behaviors were collected on all subjects. Pain perceptions showed positive significant correlations with depression (P = 0.01) and total percent of NK cells (P = 0.04). Depression and health behaviors were negatively correlated (P = 0.01). Positive associations were observed for depression and 2 PHA mitogen levels (P < 0.05). Comparison of group means, controlling for depression and health behaviors, showed that chronic pain patients' immune function, as measured by the combined NK effector to target (E:T) cell ratio levels, was significantly higher than the no-pain comparison group. Pain perceptions may have a deleterious effect on enumerative NK cell measures and depression levels. However, people with chronic pain may not experience attenuation in functional immune parameters. PMID- 12886668 TI - Novel predictors of acute coronary syndrome outcomes. AB - Morbidity and mortality rates among patients with acute coronary syndrome (ACS) remain high, and it is difficult to determine which patients will progress satisfactorily and which patients will have poor outcomes. Research has indicated that the inflammatory process is involved in coronary disease. There is great interest within the research community in determining if inflammatory markers could be used to determine the severity of the disease process and therefore serve as a prognostic tool for clinicians. This article describes the inflammatory process in ACS and provides a review of the current diagnostic studies of endothelial inflammatory markers (EIMs) in heart disease. Although research results of EIMs have not all been significant in determining outcomes, there is some evidence that they may be more specific than other generalized inflammatory markers, such as C-reactive protein. Future research of EIMs in patients with ACS might provide evidence of easy-to-measure and economically feasible markers that are sound prognosticators. PMID- 12886669 TI - Compliance with antihypertensive therapy after renal artery stenting. AB - Hypertension is an important clinical endpoint after renal artery revascularization for renal artery stenosis (RAS). Medication compliance is a critical determinant of blood pressure control. Although factors influencing compliance are known in essential hypertension, they have not been evaluated in studies investigating renal artery revascularization. The aim of this study was to assess the determinants of compliance to antihypertensive therapy in patients with RAS following renal artery stent placement (RASP). A cross-sectional study evaluating blood pressure, antihypertensive medications, quality of life, compliance, and determinants of compliance to antihypertensive therapy was undertaken in 112 patients undergoing RASP. Additionally, cardiovascular risk factors, antihypertensive medications, and cardiovascular history were reported. Self-reported compliance was 79% +/- 24% (scale of 0% [none] to 100% [complete] compliance) in patients after RASP. Determinants of compliance by multivariate analysis included physical symptoms, which correlated negatively to compliance and included loss of appetite (r2 = 0.26, P < 0.0001), dizziness (r2 = 0.06, P < 0.01), and cough (r2 = 0.03, P < 0.05). Systolic blood pressure correlated positively with compliance (r2 = 0.03, P < 0.05). The number or class of antihypertensive medications did not influence compliance. Patients' physical symptoms and level of systolic blood pressure, rather than the number or class of medications, influence compliance in patients with continued hypertension after RASP. Attention to physical symptoms may help to improve blood pressure control in this population. PMID- 12886670 TI - The effects of endurance exercise and vitamin E on oxidative stress in the elderly. AB - To examine the effects of exercise and vitamin E supplementation on oxidative stress in older adults, 59 participants, age 76.3 +/- 4.2 years, were randomly assigned to 1 of 4 groups: an exercise group taking placebos (EGP) or vitamin E (EGE) or a sedentary group taking placebos (SGP) or vitamin E (SGE). Measures included weight, VO2max, blood pressure (BP), and serum concentrations of vitamin E and lipid hydroperoxide (LOOH). At the end of the 16-week trial, the EGP and EGE had significant increases in VO2max and significant decreases in resting BP, weight, and LOOH concentrations (P < 0.05). The SGE had significant decreases in LOOH and BP (P < 0.05). There were no significant changes in the SGP (P > 0.05). The results suggest that endurance exercise in combination with vitamin E reduces oxidative stress, improves aerobic fitness, and reduces BP and weight in older adults. Even sedentary participants who take vitamin E may reduce oxidative stress and lower BP. PMID- 12886671 TI - Impact of sex and gender on irritable bowel syndrome. AB - Irritable bowel syndrome (IBS) is a common functional bowel disorder characterized by abdominal pain and change in defecation pattern. This review addresses the topic of possible sex (genetic, biological) and gender (experiential, perceptual) differences in individuals with and without IBS. Several observations make the topic important. First, there is a predominance of women as compared to men who seek health care services for IBS in the United States and other industrialized societies. Second, menstrual cycle-linked differences are observed in IBS symptom reports. Third, women with IBS tend to report greater problems with constipation and nongastrointestinal complaints associated with IBS. Fourth, serotonin (5-HT3) receptor antagonist and 5-HT4 partial agonist drugs appear to more effectively diminish reports of bowel pattern disruption in women with IBS as compared to men. This review examines sex and gender modulation of gastrointestinal motility and transit, visceral pain sensitivity, autonomic nervous system function, serotonin biochemistry, and differences in health care-seeking behavior for IBS. PMID- 12886672 TI - [Today neuro-ophthalmology]. PMID- 12886673 TI - [Pathogenesis of idiopathic macular hole --trend, opinions, discussions]. AB - Pathogenesis of idiopathic macular hole is very discussed subject now, the macular changes result after the vitreal surface changes of the macular region. The use of SLO and OCT has renewed interest in the early stages of macular hole and has shown that a foveal cystoid space or pseudocyst is the primary lesion in macular hole formation, together with incomplete perifoveal posterior hyaloid detachement. PMID- 12886674 TI - [Post-operatory biometry and refraction results estimated and refraction surprises--clinical study]. AB - Preoperative biometry has a major influence on the success or failure of IOL implantation procedures. The paper wishes to demonstrate the significance of this manoeuvre for the refraction and visual outcome and for the postoperative satisfaction rate of the patients. The clinical study has rulled out for one year period and a number of 239 patients suffered a IOL implantation procedure (cataract or lens extraction for refractive purposes). The preoperative biometry was always necessary. We used one of the existing IOL power calculation formula (SRK II, SRK T, Holladay). The surgeon decided about the IOL power, always after discussing with the patient their postoperative refractive expectations. The postoperative refraction was, in the majority of cases, the estimated refraction and the errors were similar with those from the literature. Refractive surprises of several diopters were registered in high myopic and hyperopic eyes or in cases of coexisting ocular diseases which have modified the anatomical parameters of the eye. In conclusion, small errors in calculation can have large effects on visual outcome. This problem will be solved in the future, because the field is evolving rapidly driven by a new understanding of the anatomy of the eye and new hardware for measuring that anatomy. PMID- 12886675 TI - [Fluorescein angiography in diabetic retinopathy]. AB - Fluorescein angiography plays an important role in diagnosis, classification and management of diabetic retinopathy. He paper presents angiographic aspects of some patients from "LASER OPTICS" office. An accurate and early diagnosis, and also the evaluation of laser treatment need fluorescein angiography as a useful method in the follow-up of diabetic patient. PMID- 12886676 TI - [Ocular onset in general serious diseases]. AB - The paper intends to show the possibility to establish the diagnosis of some severe general diseases beginnning with ocular onset. Thus, we present tow clinical cases in which the association of careful complete clinical examination and paraclinical investigations, during the hospitalization for ophthalmological therapy, leaded to unexpected findings of vital risk diseases, without any other sign at that time. PMID- 12886677 TI - [Choroid metastasis consequence of breast neoplasm]. AB - The paper presents a clinical case of a 46 years old woman with metastatic carcinoma the leading point being breast and the therapeutical modalities taken in this situation. PMID- 12886678 TI - [Pigmentary tumors of the conjunctiva in children: analysis of surgical decision]. AB - Pigmentary tumors of the conjunctiva are rare in children, but they deserve special attention as, under some circumstances, they can question the quo ad vitam prognosis. The management of these tumors is controversial because the clinical diagnosis is uncertain, not reliable for a long term evolution prognosis. In this study, after a short review of the literature, the authors present their own experience in the field (47 cases during the last 10 years, out of which 18 underwent surgery) and they try to develop an algorithm of surgical decision based on age-related factors, clinical appearance, aesthetic factors and on the possibility of establishing a good relationship with the patient and the family. PMID- 12886679 TI - [Congenital retinal macro-vessel]. AB - This is to show the case of a 40 year old patient, where a congenital retinal macro-vessel was found, the peculiarities of which are increased calibre, intersection of the macular area, arteriovenous shunts and lack of functional disturbances. The disease is rare and its lesional and functional stability is remarkable. PMID- 12886680 TI - [Postoperative complications in cataract surgery]. AB - The paper is a statistical retrospective trial that evaluates the prevalence of postoperative complications of the cataract surgery in the IInd Department of Ophthalmology Timisoara. The parameters of interest were the type and the frequency of the postoperative complication encountered after the extracapsular cataract extraction (ECCE) followed by a PMMA intraocular lens (IOL) implantation. The most frequent early postoperative complications were endoepithelial comeal edema (20.8%), inflammatory complications (uveitis) (8.72%), hemorrhagic complications (4.02%), and postoperative rise of IOP (8.05%). Out of late complication, we encountered cystoid macular edema (1.34%), bullous keratopathy (1.34%), IOL malposition (0.67%) and secondary glaucoma (1%). PMID- 12886681 TI - [Treatment difficulties in ocular toxoplasmosis]. AB - PURPOSE: To determine the increased incidence of ocular infection with Toxoplasma gondii, the variety of ocular manifestations and therapeutical problems of this potential blind disease. We analyzed 22 patients hospitalized in our clinic in last 2 years at whom clinical diagnosis was confirmed by positive serological tests. Is discussed the correlation between serum level antibody(IgG and IgM) and ocular manifestations and the opportunity of beginning specific treatment depend on these two parameters. CONCLUSIONS: The incidence of ocular toxoplasmosis is increasingly, the ophthalmologist must be open on atypical manifestations of this disease. The treatment remains controversial, we believe that it must be specific and correlated with ocular lesions evolutions and serum antibody level in time. PMID- 12886682 TI - [Anterior uveitis in seronegative spondylarthropathies]. AB - PURPOSE: To study the implication of seronegatives spondylarthropaties in anterior uveitis ethiology. We analyzed 51 patients with rheumatologic disease related uveitis hospitalized in our clinic in last 4 years. We discussed sex and age group repartition of patients, type of rheumatologic disease, recurrencies, complications and patients evolutions under treatment. CONCLUSIONS: Seronegatives spondylarthtropaties related uveitis represent 26% from 208 patients with uveitis household in our clinic in period mentioned above. They have a slow evolution with multiples recurrences and complications, requiring long period of treatment and follow-up. Visual prognosis is reserved because of multiples recurrences which accentuate morfofunctional deficit every time. PMID- 12886683 TI - [Association between iris examination and general pathology]. AB - PURPOSE: The research proposal is to evaluate the association between certain irian signs and general pathology of studied patients. METHOD: There were studied 57 hospitalized patients; there was taken over all their iris images, which were analyzed through iridological protocols; in the same time the pathology of these patients was noted from their records in the hospital, concordant with the clinical diagnosis; all these information were included in a database for a computerised processing. RESULTS: The correlations resulted from, shows a high connection between the irian constitution establish through iridological criteria and the existent pathology. CONCLUSIONS: Iris examination can be very useful for diagnosis of a certain general pathology, in a holistic approach of the patient. PMID- 12886684 TI - [Hyper and hypo correction in strabismus surgery]. PMID- 12886685 TI - In-vivo investigations on dye-enhanced photothermal tumor therapy with a naphthalocyanine derivative. AB - Chromophore-enhanced photothermal therapy involves the application of an exogenous chromophore in combination with irradiation, using an appropriate wavelength, exposure duration and sufficient irradiances. The chromophore palladium(II) octabutoxynaphthalocyanine (PdNc(OBu)8) accumulates at satisfactory concentrations and with good selectivity between both tumor and muscle and tumor and skin in tumor-bearing mice. In an attempt to thermally damage tumor tissue with concurrent sparing of adjacent normal tissue, the potential of PdNc(OBu)8 for photothermal therapy was investigated. Using a Balb/c mouse model with subcutaneously implanted EMT6 adenocarcinoma, 90-100 hours after intraperitoneal application of PdNc(OBu)8, the tumor and surrounding tissue were irradiated with a 826nm continuous-wave diode laser. The thermal effects on tumor and normal tissue were evaluated histologically. Our results indicate that after PdNc(OBu)8 administration and tumor irradiation using 5W/cm-for 100 seconds, pronounced selective heating of the tumor was achieved in mice, while in control animals merely an unspecific and marginal overall increase in temperature over the entire irradiation area was observed. Histological evaluation of treated areas indicated that the PdNc(OBu)8-targeted tumor tissue showed severe thermal damage while peripheral tissue like skin and muscle remained largely unaffected. This study shows the potential of creating localized thermal effects by using PdNc(OBu)8 and continuous-wave light for chromophore-enhanced photothermal therapy. PMID- 12886686 TI - New glycosylated porphyrins for PDT applications. AB - A good sensitizer for photodynamic therapy (PDT) should be a very selective reagent, which should fastly eliminate from healthy tissues. Furthermore it should have a strong absorption in the red light and good energy transfer properties to molecular oxygen to produce singlet oxygen. All these specifications imply that many second generation photosensitizers (porphyrins, chlorins, bacteriochlorins...) have been modified in order to improve their properties, but however, few have received the approval by regulatory authorities. One way to increase the selectivity of a photosensitizer is coupling it to a vector. Carbohydrate-bound porphyrins were found to be of great interest because of the specific affinity of particular carbohydrates for some tumour cells, the increasing of plasmatic life time and solubility. In this study, we report the synthesis and the photophysical properties (absorption, fluorescence, singlet oxygen formation) of new glycosylated porphyrins. Then, in vitro photocytotoxic properties were evaluated on the human colon adenocarcinoma cell line HT29. PMID- 12886687 TI - Photochemical internalization (PCI)--a novel technology for release of macromolecules from endocytic vesicles. AB - The utilisation of macromolecules in therapy of cancer and other diseases is becoming increasingly relevant. Recent advances in molecular biology and biotechnology have made it possible to improve targeting and design of cytotoxic agents or DNA complexes for clinical applications. To achieve the expected biological effect of these macromolecules in many cases internalization to the cell cytosol is crucial. A number of different methods for internalization of membrane impermeable molecules has been established, including electroporation, liposome fusion, antibodies/targeting ligands as protein carriers and the utilisation of various types of vectors such as cationic polymers and viruses, for gene therapy. Although new delivery systems have improved the cellular uptake of macromolecules, tissue penetration, cellular uptake and efficient transfer of the molecules into the cytosol of the target cell are still fundamental obstacles. At an intracellular level, the most fundamental obstruction for cytosolic release of the therapeutic molecule is the membrane-barrier of the endocytic vesicles. Photochemical internalization (PCI) is a novel technology for release of endocytosed macromolecules into the cytosol. The technology is based on the use of photosensitizers located in endocytic vesicles that upon activation by light induce a release of macromolecules from their compartmentalization in endocytic vesicles. PCI has been shown to potentiate the biological activity of a large variety of macromolecules and other molecules that do not readily penetrate the plasma membrane, including proteins, peptides, and DNA delivered as a complex with cationic polymers or incorporated in adenovirus. The basis as well as the utilization of this technology will be briefly reviewed in this paper. PMID- 12886688 TI - [Histopathologic aspects of degenerative and inflammatory disease of cornea]. AB - OBJECTIVES: Maintaining the transparency of the cornea is the most important factor in keeping its optical function. The purpose of this study is to evidentiate the structural changes suffered by the cornea during different ocular diseases, thus emphasizing the importance of applying, in due time, the correct therapy. MATERIAL AND METHODS: The studied material comes from two eyeballs, obtained as a result of enucleations performed on patients with painful primary glaucoma-absolute stage, secondary glaucoma-absolute stage. For comparison, a witness group represented by eyeballs without lesions was used. The material underwent histological techniques on paraffin and performed routine coloration. RESULTS: The examination of the histological sections coming from the witness group revealed the known classical structure of the cornea. In glaucoma, the histological aspects are consequence of the ocular hypertension on the structures of the eyeball, depend on the stage of the disease, but are not influenced by the clinical form (open angle glaucoma and closed angle glaucoma). CONCLUSIONS: The obtained results indicate dystrophic, circulatory and inflammatory lesions, that can lead to the loss of the transparency of the cornea, and thus to the loss of function of the visual organ. The aspects are important for establishing a precocious diagnosis and choosing the correct therapy. PMID- 12886689 TI - [Histopathological assessment of corneal preservation]. AB - PURPOSE: To show corneal changes after corneal preservation. MATERIAL AND METHOD: There was studied 10 pig corneas: a fresh cornea; two corneas stored in wet chamber at 4 degrees C, 24 and 48 hours; seven corneas stored in Inosol at 31 degrees C, 24 hours, 7, 10, 14, 18, 20 and 22 days. RESULTS: We obtained images of every cornea; we followed epithelial, stromal and endothelial changes. CONCLUSIONS: 1. During corneal preservation there are progressive histological changes, early in epithelium and stroma, later at endothelial level. 2. The safe preservation period in wet chamber at 4 degrees C is 24 hours; the changes found after 48 hours are irreversibile. 3. The safe preservation period in Inosol at 31 degrees C is 14 days. PMID- 12886690 TI - Synthetic metallo-porphyrins and selection criteria for PDT. AB - The aim of this paper is to analyse the some aspects about the main selection criteria of some metallo-porphyrins for PDT; purity, solubility, singlet oxygen generation and photostability. Also, some clinical applications of some porphyrins are evaluated. The PDT effect of some metallo-porphyrins on Murine Lymphoma EL-4 cells (as experimental model for photodynamic therapy studies), on Herpes Simplex Virus, on human brain cells, in ophthalmology, are discussed in this review. PMID- 12886691 TI - [The eye in Ion Tuculescu's paintings]. AB - The painter Ion Tuculescu was placed through his painting, in the current of the abstract expressionism. He endowed his pictural skill with a personal vision of life and people, through introduction the eye like a stylized aspect and its loading with spiritual meanings. The eye is practically in every picture of the painter, constituting a philosophy of his interior experiences and attitude given the existence. The subject painting has not figurative plans but mental impulses through the artist constitutes a new visual language. PMID- 12886692 TI - [Jaffe syndrome (primitive syndrome with retraction of the internal macular limiting membrane)]. PMID- 12886693 TI - Testing for micro-albuminuria in diabetes mellitus. PMID- 12886694 TI - Iodine deficiency disorders in South Africa. AB - Iodine is an essential micronutrient, but iodine deficiency remains common worldwide, including many African countries. In South Africa endemic goitre was first noted in the 1920s. Several subsequent surveys have indicated its widespread persistence, despite the limited availability of iodised salt. The significance of nutritional iodine deficiency lies in the associated iodine deficiency disorders, which follow from maternal and fetal hypothyroidism. The extreme consequence is endemic cretinism, but much more prevalent are lesser degrees of intellectual and neurological deficits, with a potential reduction of intelligence scores in affected communities of more than 10%. Such severe consequences of iodine deficiency probably affect a significant proportion of South Africans. The control of iodine deficiency should be easy and inexpensive. Since the December 1995 legislation, all salt manufactured in South Africa must contain potassium iodate. Such supplementation is likely to cause a small increase in auto-immune thyroid disorders. This negative effect is slight in comparison with the potential benefit to millions. PMID- 12886695 TI - Dietary iodine deficiency in South Africa. Surveys before the introduction of universal salt iodisation. AB - OBJECTIVE: To survey iodine nutritional status in several geographically separated communities in South Africa. DESIGN: In an initial study total goitre prevalence (TGP) was correlated with urinary iodine concentration (UIC) in some 300 primary school children in a single district. Thereafter only UIC was surveyed in children from 5 additional communities. RESULTS: In the initial survey in Mpumalanga TGP was 74.2% (23.4% visible) and the median UIC was 15.6 micrograms/l, both data indicating severe iodine deficiency in this district. Median UIC values indicated mild to severe iodine deficiency in districts in the Northern Province, moderate deficiency in the Eastern Cape, and mild deficiency in Soweto, Gauteng. Only non-black African children in Johannesburg were iodine replete (UIC > 100.0 micrograms/l.) CONCLUSION: Before the introduction of compulsory iodisation of salt in December 1995, dietary iodine deficiency was widespread in South Africa. PMID- 12886696 TI - Assessment of growth--the role of knemometry. PMID- 12886697 TI - Salt sensitivity is not associated with hyperinsulinaemia in a sample of rural black Zimbabweans. AB - OBJECTIVES: To investigate the relationship between salt sensitivity and hyperinsulinaemia in rural black African subjects. DESIGN: An intervention study where 27 subjects were divided into two groups; group 1 was initially salt loaded (300 mmol Na+/day), while group 2 was salt restricted (25 mmol Na+/day), each for 4 days, after which a cross-over study was done. SETTING: Chidamoyo, a rural area 383 km north of Harare, Zimbabwe. SUBJECTS: Twenty-seven rural volunteers (16 women, 11 men). OUTCOME MEASURES: Systolic and diastolic blood pressures, salt sensitivity, insulin and glucose levels, body mass index and mean arterial pressure. RESULTS: Mean arterial pressure, which was 91 +/- 2 mmHg on a low-salt diet, increased significantly (P < 0.01) to 105 +/- 3 mmHg on high-salt diet in the salt-sensitive subjects. In the same salt-sensitive subjects, the fasting insulin level was 8.4 +/- 0.8 microU/ml on a low-salt and 6.1 +/- 1.0 microU/ml on a high-salt diet. The difference was not statistically significant. CONCLUSIONS: Although salt pressor sensitivity was demonstrated in the subjects, there was no accompanying increase but rather a decrease in fasting insulin levels, suggesting that in the short term, salt sensitivity and hyperinsulinaemia are not linked in raising blood pressure in this sample of rural Zimbabwean subjects. PMID- 12886698 TI - Micro-albuminuria in diabetes mellitus--significance and screening. PMID- 12886699 TI - Antiviral treatment of human immunodeficiency virus infection: update for 2003. AB - Highly-active Antiretroviral Treatment (HAART) nowadays consists of 21 approved drugs in the USA. The medications belong to four classes: (1) Nucleoside/nucleotide analogues, (2) Non-nucleoside reverse transcriptase inhibitors, (3) Protease inhibitors, and (4) a fusion inhibitor. At least three effective drugs are required to reduce the viremia, aiming at nondetectable, and thus, raise the immune reserve, i.e. the CD4 lymphocyte count. HAART effectively extends the life of patients infected with HIV, in essence converting a fatal disease into a chronic infection, as eradication of HIV is not yet possible. The laurels of HAART are shadowed by its toxicity. Non-compliance with treatment is also an important contributor to its failure. HAART is continuing in development, change, benefiting the patients affected with HIV/AIDS, HCV and HBV. PMID- 12886700 TI - Sentinel lymph node examination by frozen section in breast cancer patients. AB - Our study indicates that sentinel node analysis by frozen section at the time of primary surgical treatment for invasive breast cancer is a reasonable and prudent effort. No cost analysis study is projected in this paper, but the risk/benefit to the patient as well as cost savings would seem to be apparent and significant. PMID- 12886701 TI - 2003 update on the Hypertension Initiative of South Carolina. Bringing South Carolina from "worst to first" in cardiovascular health. AB - Hypertension and its related comorbidities continue to drain South Carolina of approximately $9 billion dollars a year in direct medical costs and indirectly through lost productivity. Improving control rates of blood pressure and associated cardiovascular risk factors in the primary care physician's office is a fundamental and crucial step to decrease the very high incidence of stroke and cardiorenal diseases. By monitoring prescribing patterns through the Initiatives' data feedback program, providing evidence-based management approaches through educational seminars, and by applying improved treatment protocols, physicians can have a profound impact on outcomes. The growing collaborative partnership spawned by the Hypertension Initiative now includes primary care providers, Hypertension Specialists in the local community, and the ASH Carolinas-Georgia Chapter. The growth and impact of the partnership is facilitated by a dynamic data auditing and feedback program that provides the basis for constructive change focused on optimizing cardiovascular risk factor control in patients across the State. By continuing to make progress in addressing the control of blood pressure and associated cardiovascular risk factors through a growing collaborative partnership with primary care providers statewide, South Carolina can move from worst to first in cardiovascular health. In the future, we hope to add a dynamic health promotion program to active disease management efforts. In the process, South Carolina can move from a leader in cardiovascular disease to a model of cardiovascular health. PMID- 12886702 TI - Incidence of astrocytoma in South Carolina: follow-up study. AB - As a result of a statewide cancer registry in South Carolina, we are able to conclude that there is a higher probability for occurrence of astrocytoma in the Upstate when compared to the remainder of the state. Hopefully these results will foster additional research into potential etiologies for this phenomenon. PMID- 12886703 TI - The general internist and the role of "the hospitalist". PMID- 12886704 TI - Drug wars: saying no. PMID- 12886705 TI - Class actions for needle sticks are still alive and well. PMID- 12886706 TI - Nurse does not get promotion: sex discrimation suit fails. Case on point: Longstreet v. Holy Spirit Hospital, 2003 WL 21205881 Fd.3d-PA. PMID- 12886707 TI - NY: nurse develops heel spur-files comp. claim: is heel spur an occupational disease? PMID- 12886708 TI - AR: RN has non-work-related back surgery: temporary total disability comp. benefits denied. PMID- 12886709 TI - Nurse-expert fails to 'go to bat' in testifying for plaintiffs. Case on point: Ross v. Redding Medical Center, 2003 WL 21246105-CA. PMID- 12886710 TI - Clarity of vision. PMID- 12886711 TI - Nasal reconstruction. AB - Nasal defects commonly seen by plastic surgeons result from trauma, burn injury, or tumor resection. While nasal reconstruction is one of the oldest plastic surgery endeavors, techniques continue to evolve and be modified. Grafts and local flaps are used in smaller defects. Larger and complex defects are best reconstructed following the aesthetic unit principle. These defects also require replacement of all lost tissues to provide nasal lining, skeletal support, and skin coverage. Careful analysis of the defect and reliance on these general guidelines will allow for less obvious nasal reconstruction and a more natural appearance and function. PMID- 12886712 TI - Leprosy in a Mexican immigrant. AB - A new diagnosis of borderline lepromatous leprosy was established in a man who had immigrated to Kentucky from Mexico. He was placed on a World Health Organization treatment regimen consisting of dapsone, clofazimine, and rifampin. The biology of leprosy, its diagnosis, treatment, and worldwide impact are reviewed. Because of the potential for highly mobile populations to export endemic diseases, Kentucky physicians must expand their lists of differential diagnoses. PMID- 12886713 TI - Ethics and the gift. PMID- 12886714 TI - Medicare: industry benchmark or tool of collusion? PMID- 12886715 TI - CMS proposes new Medicare enrollment requirements. PMID- 12886716 TI - Following a road map to success. PMID- 12886717 TI - Forming an attachment. Network-attached storage for electronic document management. PMID- 12886718 TI - Renewing our social contract. Collective accountability the key to improving our professional image. PMID- 12886719 TI - Business office basics. Case study: fundamentals yield improved collections. PMID- 12886720 TI - Use the code. A code of conduct may be your best compliance tool. PMID- 12886721 TI - Partnership for care. Group practices see disease management as the future for chronic illness. PMID- 12886722 TI - Stay safe. Medical mistakes make headlines. AB - Medical mistakes in ambulatory settings are not isolated events, and most errors are avoidable. Your patients can be safer--and your practice can stay off the evening news--if you implement systems and processes that will minimize errors. PMID- 12886723 TI - On alert. Reducing risk in the medical office. AB - A comprehensive review of medical office systems can decrease risk in the practice. Education and training of employees are key to this goal. When staff follow policies and protocols consistently, they help protect patients and patient rights. Checklists for front- and back-office employees can help a medical group maintain protections and deliver quality care. PMID- 12886724 TI - Pick and choose. Selecting billing and scheduling software. AB - HIPAA's regulations are spurring many medical groups to change their billing and/or scheduling software. Here is a step-by-step process to evaluate your practice's needs, find a vendor and figure the time the process will take. Checklists can assist you at each stage of the process. PMID- 12886725 TI - Physician assistants have been a part of Hawaii's health services delivery system. PMID- 12886726 TI - Duty to third parties: a new worry for doctors. AB - The Hawaii Supreme Court ruled on June 10, 2002, that physicians might be liable to non-patient third parties if they fail to warn their patients regarding a medication's adverse effect on driving. Conceivably, this liability may also extend to physicians who fail to inform their patients and/or the Department of Motor Vehicles of medical conditions that affect operating a vehicle safely. Physicians must be cognizant of every medication's impact on driving ability, inform their patients of these adverse effects, and should consider asking their patients to stop driving if the risks are substantial. PMID- 12886727 TI - Case report: an ingestion of Hawaiian Baby Woodrose seeds associated with acute psychosis. AB - An 18 year old male ingested Hawaiian Baby Woodrose seeds (containing the hallucinogen lysergic acid) and required hospitalization for psychotic behavior. This is the second documented case of an acute psychosis ascribed to ingestion of this plant, and it is suggested that Hawaiian Baby Woodrose ingestion should be present in the differential diagnosis when dealing with the acutely psychotic young adult patient. PMID- 12886728 TI - End-of-life care curriculum for internal medicine residents at the John A. Burns School of Medicine, University of Hawaii. PMID- 12886729 TI - Statewide comprehensive cancer control in Hawaii. PMID- 12886730 TI - Smokeless tobacco as a smoking cessation strategy. PMID- 12886731 TI - Narcotics for diabetic neuropathy. PMID- 12886732 TI - ACNP testifies at federal hearings about barriers to practice. PMID- 12886733 TI - Business savvy and the NP role. PMID- 12886734 TI - Certification for practicing NPs. PMID- 12886735 TI - New headache cause for concern. PMID- 12886736 TI - When competition is queen. PMID- 12886737 TI - Insulin therapy today. Focusing on the basal-bolus balance. PMID- 12886738 TI - Assessment and management of pressure ulcers in the elderly. Principles of care. PMID- 12886739 TI - Animal-inflicted hand wounds. Treat early and aggressively. PMID- 12886740 TI - Recalcitrant venous stasis ulcers. A guide for nurse practitioners. PMID- 12886741 TI - Chronic hepatitis C. Treatment and side effect management. PMID- 12886742 TI - Skin manifestations in the newborn. Differentiation and diagnosis. PMID- 12886743 TI - Patient information. Probiotics: getting a little culture. PMID- 12886744 TI - Clarifying the clinical doctorate as the degree gains attention, educators seek simplification. PMID- 12886745 TI - From sandwiches to center stage. Peanuts pack a powerful nutritional punch. PMID- 12886746 TI - [State of the art in surgery of gastric cancer]. PMID- 12886747 TI - [From molecular biology to new treatment approaches to colorectal cancer: basic research, experimental trials and surgical implications]. AB - The Authors review the natural history of colorectal cancer from the point of view of molecular biology and genetics from aberrant crypts foci and familiar adenomatous polyposis to hereditary non polyposis colon cancer and sporadic colorectal cancer. They carry out international literature about basis knowledges, experimental trials and personal studies. Up to day traditional colorectal cancer surgical treatments and adjuvant or neoadjuvant pharmacological therapy cannot be modified, nevertheless "new drugs generation" known as signal transduction inhibitor could, in the future, prove to be an effective cancer treatment. The Authors highlight recent experimental clinical trials probably able to prevent sporadic colorectal cancer spreading and precursor evolution. PMID- 12886748 TI - Bile leak from the accessory biliary duct following laparascopic cholecystectomy. AB - Anatomists and surgeons have described the presence of accessory biliary ducts between the liver and gallbladder. Bile leakage from accessory duct following laparoscopic cholecystectomy (LC) is an unusual post-operative complication. Aim of the study was to assess its incidence, the intraoperative methods helpful for notice the anatomical anomaly and the impact of endoscopic procedure as a suitable treatment. From January 1997 to September 2002, 185 patients underwent LC for symptomatic cholelithiasis in our surgical department. Post-operative bile leakage from accessory biliary duct occurred in two patients (1%): one case from the liver bed of gallbladder (duct of Luschka) and one case from an aberrant cholecystohepatic duct entering Hartmann's pouch. One patient underwent open celiotomy because of unavailability of endoscopic retrograde cholangiopancreatography. The other patient was successfully treated by endoscopic sphincterotomy and nasobiliary tube placement. By careful dissection, accessory ducts were noticed and clipped in three other patients with overall incidence of 2.7%. Meticulous laparoscopic technique aimed to careful recognize all structures during LC is the main policy to contain biliary injury within its nadir incidence. Depending of availability, endoscopic sphincterotomy and nasobiliary drainage allow diagnosis and treatment of bile leakage, preserving the effectiveness of laparoscopic procedure. PMID- 12886749 TI - [Inflammatory tumors of the cecum in emergencies: surgical considerations and case report]. AB - A cecal mass of uncertain etiology encountered during surgery for presumed appendicitis is a dilemma for the surgeon. The differential diagnosis of an unsuspected ileocecal mass must include neoplasm, diverticular disease, inflammatory bowel disease and severe appendicitis involving the ileocecal region. Right hemicolectomy is the gold standard for the treatment of the unsuspected ileocecal mass. The Authors report a case of inflammatory tumour of caecum secondary to appendicitis in a young adult. PMID- 12886750 TI - [Intraoperative cholangiography during laparoscopic cholecystectomy: selective or routine?]. AB - Since its presentation by Mirizzi in 1931, the role of intraoperative cholangiography (ICHO) has been controversial and has become an argument even more disputed with the introduction of laparoscopic cholecystectomy (VLC) in 1988. The Authors reviewed their experience to determine the most appropriate use of ICHO during VLC on the basis of a retrospective analysis of cases of selective ICHO. From December 1991 to January 2001, 597 patients, 552 elective procedure and 45 emergency procedure, were reviewed. Of 552 patients 62 presented with at least one diagnostic criterion for symptomatic gallstone disease and were treated by means of ERCP completed with endoscopic sphincterectomy (ES) when a stone of the common bile duct was found, while the remaining 490 patients underwent VLC; a total of 10 ICHO were performed, two of which in the ERCP group and 8 in the VLC group. The 45 patients treated in emergency underwent VLC; in 43 cases ICHO was performed. Of all patients, there were 2 cases of common bile duct injuries (0.33%) and in both cases ICHO was not performed. A cholangiogram added 27 min to the average duration of surgery. On the basis of both the literature and Authors' experience, it can be stated that the routine use of ICHO is not useful to reduce bile duct injuries, while it significantly increases the cost of the surgical procedure due to the increase of average operative time. The use of ICHO seems to be effective to demonstrate clinically unsuspected choledocholitiasis, although, at present, the real clinical advantage deriving from the detection of these stones is not clear. The Authors conclude that further prospective, randomized studies are necessary to assess the precise role of ICHO with regard to VLC. PMID- 12886751 TI - An atypical presentation of Crohn disease in the elderly. A case report and Literature review. AB - The Authors present a case of an insidious onset of Crohn Disease (CD) in an elderly patient. Diagnosis complicated by extraintestinal manifestation properly of old age could be delayed and often made after surgery on the histological specimen as in our case. CD is uncommon as primary manifestation in old age, often unsuspected, incorrectly diagnosed and in many case the clinical features may lead to late diagnosis. Differential diagnosis of CD in elder people with fever, diarrhoea and abdominal pain is difficult and other symptoms affecting intestinal tract can closely mimic CD symptoms, although the pattern of clinical presentation in older patient resemble those in younger. PMID- 12886752 TI - [Importance of nursing in the prevention of complications of totally implantable venous access devices]. AB - The Authors report a case of a patient submitted to chemotherapy that has a complication of its totally implantable venous access device (TIVAD) due to incorrect nursing. The symptoms related to the extravasation of drugs were due to the lateral lesions of the catheter with Huber needle. After X-ray examination the therapy was ablation of the non-functioning device and implant of a new TIVAD. The Authors conclude about careful nursing of the TIVAD in order to avoid further psychological problems in cancer patients. PMID- 12886753 TI - [Idiopathic gynecomastia: our experience]. AB - The Authors reporting their experience, discuss some concepts about physiologic evolution of male and female breast. They distinguish between real and false gynaecomastia; stressing the causes of abnormal development of male breast and morphopathological characteristics of gynaecomastia. Careful diagnostic protocol is necessary for a therapeutic approach based on traditional surgery or liposuction: it depends on prevalence of glandular or adipose breast's tissue. PMID- 12886754 TI - [Sacrococcygeal pilonidal sinus disease. Treatment by "open" and "closed" technique: personal experience]. AB - The Authors report their own experience concerning the treatment of the sacrococcigeal pilonidal sinus disease. Results obtained with the "open" and "closed" techniques were compared. Although results were similar in terms of complications and relapses, the closed technique is to be preferred, because it allows a more rapid return to working activities. In the light of the new trends of sanitary economy a modulated treatment of the disease is suggested. PMID- 12886755 TI - [Color Doppler ultrasound in acute mesenteric ischemia: 2 case reports]. AB - Acute mesenteric insufficiency is a surgical emergency with a difficult methodological approach. Its high mortality is mainly due to delay in the correct diagnosis. In turn this is due to the lack of specificity, especially in the early-middle phase of the clinical presentation, and of the laboratory data and abdominal radiographic findings. Echo-doppler ultrasound promises to be more accurate than conventional ultrasound in the diagnosis of visceral ischemia and may help to identify those patients who may require angiography. Two women presenting atrial fibrillation, abdominal pain, leukocytosis, hyperamylasemia were accepted in October 2000 ad March 2001. US colordoppler was performed 12 hours after admittance demonstrating a "stop" of enhancement of the superior mesenteric artery in one case and absence of enhancement in the other. PMID- 12886756 TI - [Post-mastectomy breast reconstruction by tissue expanders: technical note]. AB - The aim of post-mastectomy breast reconstruction by tissue expanders is the mammary symmetry. The Authors report a personal experience in immediate (55 pts) and delayed (12 pts) breast reconstruction with implants. The technique is simple and safe with good aesthetical and functional results. PMID- 12886757 TI - Renal disease. Techniques and protocols. Preface. PMID- 12886758 TI - Standards of animal care in biological experiments. PMID- 12886759 TI - Models of polycystic kidney disease. PMID- 12886760 TI - Rat models of the metabolic syndrome. PMID- 12886761 TI - Models of glomerulonephritis. PMID- 12886762 TI - Functional studies of the kidney with magnetic resonance imaging. PMID- 12886763 TI - Use of radionuclides to study renal function. PMID- 12886764 TI - Intravital videomicroscopy. PMID- 12886765 TI - Confocal and two-photon microscopy. PMID- 12886766 TI - Atomic force microscopy in renal physiology. PMID- 12886767 TI - Freeze-fracture analysis of renal-epithelial tight junctions. PMID- 12886768 TI - Organ culture of intact metanephric kidneys. PMID- 12886769 TI - The ureteric bud. Tissue-culture approaches to branching morphogenesis and inductive signaling. PMID- 12886770 TI - Studies of cell lineage in the developing kidney. PMID- 12886771 TI - Transient transfection assays for analysis of signal transduction in renal cells. PMID- 12886772 TI - The study of gene polymorphisms. How complex is complex genetic disease? PMID- 12886773 TI - Laser-capture microdissection. PMID- 12886774 TI - Serial analysis of gene expression. PMID- 12886775 TI - RNA labeling and hybridization of DNA microarrays. PMID- 12886776 TI - Gene-expression analysis of microdissected renal biopsies. PMID- 12886777 TI - The use of SELDI ProteinChip array technology in renal disease research. PMID- 12886778 TI - Clearance studies in genetically altered mice. PMID- 12886779 TI - Long-term blood-pressure monitoring in unrestrained animals. PMID- 12886780 TI - In vitro studies on renin release. PMID- 12886781 TI - Mitochondrial function. PMID- 12886782 TI - Detection of cysteine S-nitrosylation and tyrosine 3-nitration in kidney proteins. PMID- 12886783 TI - Products of arachidonic acid metabolism. PMID- 12886784 TI - Methods for measurements of heme oxygenase (HO) isoforms-mediated synthesis of carbon monoxide and HO-1 and HO-2 proteins. PMID- 12886785 TI - The juxtamedullary nephron preparation. PMID- 12886786 TI - Tubuloglomerular feedback. PMID- 12886787 TI - Microdissected perfused vessels. PMID- 12886788 TI - Renal microperfusion techniques. PMID- 12886789 TI - How to design a clinical trial. PMID- 12886790 TI - Mapping the disease. PMID- 12886791 TI - Debate that "this house believes the essential drug concept hinders the effective deployment of drugs in developing countries". Introduction. PMID- 12886792 TI - The essential drugs concept is needed now more than ever. AB - The debate on access to essential medicines, which was live in the late 1970s and 1980s when strong pressure from public health advocates led the pharmaceutical industry to accept the concept, is now back on the international health policy agenda. Currently, the Essential Drugs List (EDL) forms an integral part of national drug policies in 146 countries, guiding the selection of drugs on the basis of public health relevance, efficacy, safety and cost. The essential medicines debate is back on the agenda as part of the access to medicines debate. The Trade Related Aspects on Intellectual Property Rights agreement has caused concern among public health advocates in terms of its impact on access to essential medicines. This article reviews the concept of essential medicines in the light of the changing political context and recommends that the EDL is supported as a crucial public policy tool in all countries. PMID- 12886793 TI - Patents and access to essential drugs. AB - This paper provides a brief overview of historical developments in patent law including its recent incorporation into world trade law. The impact of patents on access to essential drugs will be discussed. The relationship between intellectual property rights and the right to health will be considered. PMID- 12886794 TI - Translating the essential drugs concept into the context of the year 2000. AB - The WHO Essential Drugs List (EDL) was first published in 1977. It must continue to be adapted to a changing world with new drugs, new indications and new diseases. Medecins sans Frontieres is concerned that the eleventh list is outdated. The EDL should be stricter about its inclusion of first-line drugs and price should not be a criterion for excluding drugs from the list. Second-line drugs are complementary and their inclusion would in no way undermine the essential drugs concept. The EDL should be updated regularly and in a transparent manner, by an independent panel of experts chosen for their competence. Decisions regarding the inclusion of drugs in the list should be evidence-based, and the evidence should be available to the panel of experts long before their final sessions on updates to the EDL. The WHO not only has the challenge of defining a EDL that addresses today's epidemiological reality, but also a responsibility to assist countries in gaining access to these treatments. PMID- 12886795 TI - Proposing the motion. PMID- 12886796 TI - Opposing the motion. PMID- 12886797 TI - Reducing infant mortality in poor countries by 2015--the need for critical appraisal of intervention-effectiveness. PMID- 12886798 TI - Implementing a community-based participatory intervention to improve essential newborn care in rural Nepal. AB - The persistence of high perinatal and neonatal mortality rates in many developing countries make efforts to improve perinatal care in the home and at local health facilities important public health concerns. We describe a study which aims to evaluate a community-level participatory intervention in rural Nepal. The effectiveness of community-based action research interventions with mothers and other key members of the community in improving perinatal health outcomes is being examined using a cluster randomized, controlled trial covering a population of 28,000 married women of reproductive age. The unit of randomization was the village development committee (VDC): 12 VDCs receive the intervention while 12 serve as controls. The key elements of the intervention are the activities of female facilitators, each of whom works in one VDC facilitating the activities of women's groups in addressing problems in pregnancy, childbirth and the newborn period. Each group moves through a participatory planning cycle of assessment, sharing experiences, planning, action and reassessment, with the aim of improving essential maternal and newborn care. Outcomes assessed are neonatal and perinatal mortality rates, changes in patterns of home care, health care seeking and referral. The study also aims to generate programmatic information on the process of implementation in communities. PMID- 12886799 TI - Antenatal and perinatal strategies to prevent mother-to-child transmission of HIV infection. AB - Mother-to-child transmission (MTCT) is the dominant mode of acquisition of HIV infection for children. Each day an estimated 1600 children born to HIV-infected mothers become infected, 1500 of whom are in sub-Saharan Africa. Mother-to-child transmission can occur before, during and after delivery; overall rates of MTCT range from 15 to 35%. Risk factors for MTCT include maternal viral load levels, vaginal delivery, prematurity and breastfeeding. Approaches to reduce the risk of MTCT include reduction of maternal viral load through antiretroviral prophylaxis, avoidance of exposure through birth canal cleansing or elective caesarean section delivery and refraining from breastfeeding, and boosting the host immune system through nutritional supplementation or immunization. Substantial reductions in the risk of MTCT can be achieved with antiretroviral prophylaxis during pregnancy, delivery and in the neonatal period, both in settings with and without breastfeeding. Elective caesarean section independently decreases the risk, but is not a safe and feasible option in most high prevalence areas. Birth canal cleansing with chlorhexidine does not reduce the rate overall, but may be beneficial in a subgroup of women with prolonged duration of ruptured membranes. Micronutrient supplementation, including vitamin A, does not reduce MTCT of HIV, but may improve pregnancy outcome generally. PMID- 12886800 TI - Infant-feeding strategies to prevent post-natal HIV transmission. AB - Promotion of breastfeeding is a key component of infant health policies globally. However, maternal HIV infection is a possible contraindication for breastfeeding since it can result in mother-to-child transmission. Research into infant-feeding strategies which will promote the health of all infants but also protect against HIV transmission is handicapped by technical difficulties in determining the exact timing of when an infant becomes infected. Nevertheless, progress is being made both in defining and promoting 'safer breastfeeding' for HIV-infected women who choose to breastfeed and in supporting the use of breastmilk substitutes by women who choose not to breastfeed. A common feature of all strategies is the need to optimize maternal health and support of women by midwives and other health care providers. PMID- 12886801 TI - Importance and prevention of malaria in pregnancy. AB - Malaria in pregnancy is one of the most important preventable causes of low birthweight deliveries worldwide. It is also a major cause of severe maternal anaemia contributing to maternal mortality. It is estimated that 40% of the world's pregnant women are exposed to malaria infection during pregnancy. The clinical features of Plasmodium falciparum malaria in pregnancy depend to a large extent on the immune status of the woman, which in turn is determined by her previous exposure to malaria. In pregnant women with little or no pre-existing immunity, such as women from non-endemic areas or travellers to malarious areas, infection is associated with high risks of severe disease with maternal and perinatal mortality. Women are at particular risk of cerebral malaria, hypoglycaemia, pulmonary oedema and severe haemolytic anaemia. Fetal and perinatal loss has been documented to be as high as 60-70% in non-immune women with malaria. Adults who are long-term residents of areas of moderate or high malaria transmission, including large parts of sub-Saharan Africa, usually have a high level of immunity to malaria. Infection is frequently asymptomatic and severe disease is uncommon. During pregnancy this immunity to malaria is altered. Infection is still frequently asymptomatic, so may go unsuspected and undetected, but is associated with placental parasitization. Malaria in pregnancy is a common cause of severe maternal anaemia and low birthweight babies, these complications being more common in primigravidae than multigravidae. Preventative strategies include regular chemoprophylaxis, intermittent preventative treatment with antimalarials and insecticide-treated bednets. PMID- 12886802 TI - Anaemia prevention for reduction of mortality in mothers and children. AB - The relationship of anaemia as a risk factor for child and maternal mortality is described. Maternal case fatality rates, mainly from hospital studies vary from < 1% to > 50%. These large differences in risk were related primarily to differences in available obstetric care for women living in areas with inadequate antenatal and delivery care facilities. The relative risk of mortality associated with moderate anaemia (haemoglobin [Hb] 40-80 g/L) was 1.35 (95% confidence interval [95% CI] 0.92-2.00) and for severe anaemia (Hb < 47 g/L) was 3.51 (95% CI 2.05-6.00). Nutritional-related anaemia mortality is likely to be greater than malarial anaemia-related mortality. With good antenatal and obstetric care most anaemia-related deaths are preventable, and policies to reduce anaemia prevalence should not be divorced from efforts to provide adequate antenatal and delivery facilities for women in developing countries. In children, although mortality was increased with anaemia (< 50 g/L), the evidence for increased risk with less severe anaemia was inconclusive. A survival analysis of Malawian infants indicated that if Hb decreased by 10 g/L after 6 months of age, the risk of dying before 12 months of age increased 1.72 times. Evidence from a number of studies suggests that mortality due to severe malarial anaemia in children is greater than that due to iron-deficiency anaemia. Primary prevention of nutritional and malarial anaemia in young children could lead to reductions in child mortality. PMID- 12886803 TI - Preventing low birthweight and reduction of child mortality. AB - Surprisingly little attention is paid to birthweight improvement as a means of reducing child mortality. Half of the 10 million pre-school-age children that die each year have malnutrition as an underlying or associated cause. Furthermore, the majority of these deaths are associated with the mild and moderate forms of malnutrition, reflecting how this effect is manifest across the whole population distribution. Similar relationships are seen between birthweight and neonatal mortality, with the least risk of neonatal death occurring in children born weighing more than 3.5 kg. Child malnutrition is increasingly recognized to be largely determined during the period of fetal and infant growth, when maternal nutrition has its strongest influence. Although the effects of maternal food supplements on mean birthweight are relatively small, because of their influence across the population distribution they have great biological significance. An increase of 100 g in mean birthweight is associated with a 30-50% reduction in neonatal mortality. The programmes that need to be put in place to improve maternal nutritional status are discussed, and the programme initiatives being promoted by UNICEF to prevent low birthweight, including multiple micronutrient supplementation trials are further described. PMID- 12886804 TI - An individual-based model of Plasmodium falciparum malaria transmission on the coast of Kenya. AB - Individual-based models provide powerful tools to model complex interactions characterized by individual variability. This paper presents an object-oriented design for individual-based modelling of Plasmodium falciparum malaria transmission. Two kinds of objects, human and mosquito, that exhibit variability among individuals for parameters such as recovery and survival rates are defined. The model tracks the dynamics of human hosts and adult female mosquitoes individually. Immunity, modelled as a function of exposure history, is represented by reduced susceptibility and increased recovery rate. The model was calibrated using epidemiological data collected at 30 sites along the coast of Kenya. The sites were grouped into low, intermediate and high transmission based on mean daily human-biting rates. Simulation results show that malaria transmission was stable even in low transmission areas where the human-biting rate is approximately 0.5 bite per day. The model was used to examine the effect of infection control programmes that aim at interrupting transmission by reducing human-vector contact rates and implementing active case detection and drug treatment of infections. With this intervention, local elimination of malaria is likely with a probability of extinction of approximately 0.8 in low transmission areas. However, a small amount of immigration (> 0.3%) by infected people into the community could prevent local extinction of the parasite. In intermediate and high transmission areas, reduction in prevalence is short-lived and the probability of local elimination is low, even at high coverage levels of the intervention. PMID- 12886805 TI - Perceptions of malaria transmission, presentation and management in northern Cameroon. AB - During early 2001, 1197 adult health service users in a poor rural district of northern Cameroon were interviewed in order to discover prevalent beliefs about malaria. The survey included questions about the name of the disease, its cause and transmission, signs and symptoms, cure, and local importance as a cause of sickness and death. Interviewees showed good understanding of the signs and symptoms of malaria, its importance, and its association with the rains. Most felt that either a medical or a herbal treatment could cure malaria. Only 1% identified mosquitoes as a source of transmission. Health education to explore beliefs and to correct misconceptions should precede and accompany the introduction of technological interventions in malaria control programmes in traditional societies. PMID- 12886806 TI - Estimation of malaria transmission from humans to mosquitoes in two neighbouring villages in south Cameroon: evaluation and comparison of several indices. AB - Malaria transmission from humans to mosquitoes was assessed in two neighbouring villages in a rural area near Yaounde, Cameroon during high and low transmission seasons during 1998-2000, using several indices previously evaluated in different areas endemic for malaria but never directly compared. These indices were estimated from human parasitological data and mosquito infection rates and, for each individual, thick blood films were prepared at the same time as experimental infection of laboratory-bred mosquitoes. Among the 685 volunteers examined, the prevalence of Plasmodium falciparum gametocyte carriers was 16%, and 8% of individuals were able to infect mosquitoes. The percentage of mosquitoes that became infected by feeding on the infectious individuals was 21%. Children aged < 10 years contributed to about 75% of the infectious reservoir, although they constituted only 35% of the total population. Differences were found between the transmission seasons and the villages, and varied according to the index examined. Although there were more infectious individuals in one of the two villages, they were less infectious than those in the other village during the high transmission season. Comparative analysis of the transmission indices suggests the existence of functioning transmission-blocking immunity in one of the villages, which until now has been only hypothetically considered to play a role in malaria transmission in a natural setting. The epidemiological value of all the indices used and their accuracy in estimating the human infectious reservoir and its natural or induced variations are discussed. PMID- 12886807 TI - Trachoma, flies and environmental factors in Burkina Faso. AB - The presence of flies is one of the earliest risk factors for trachoma and it has been suggested that flies could act as vectors for transmission of chlamydiae. A national trachoma survey was conducted in 1997 in Burkina Faso to (i) study the relationship between trachoma occurrence, flies, dirty faces and some environmental factors, and (ii) investigate the role of flies in the presence of trachoma. The country was stratified into eight groups of provinces and a random sample of 30 clusters was selected in each group. All children aged < 10 years were examined for the diagnosis of active trachoma (trachomatous inflammation which was follicular and/or intense) and the dirtiness of the face and the presence of flies on the face were recorded. The children's carers were questioned about the number of baths given and daily face-washing. Household heads were asked about ownership of cattle and small ruminants. The presence of latrines, a stable, and garbage collection inside the yard was noted. Among 16,514 children examined, 27.0% had active trachoma and 3.3% intense inflammatory trachoma. Flies were present on 11.2% of children's faces and 82.4% and 19.7% of these children had active and intense inflammatory trachoma, respectively. Among the 30.2% of children with dirty faces, 70.2% had active and 10.2% intense inflammatory trachoma. In multivariate analysis, at least one daily bath showed a protective effect on both active and intense inflammatory trachoma. Face-washing twice daily was found to be significantly protective for active trachoma in some regions. A strong association was demonstrated between the presence of flies and dirty faces (odds ratio = 334, 95% confidence interval 202-546). The presence of flies on children's faces, dirty faces and trachoma appeared to be strongly associated. Although the presence of flies may be a marker of socio-economic status and is probably linked with other trachoma risk factors, our data indicated that interventions targeting fly control should be an important issue in controlling trachoma. PMID- 12886808 TI - Complications and mortality patterns due to Plasmodium falciparum malaria in hospitalized adults and children, Rourkela, Orissa, India. AB - Of 1857 Plasmodium falciparum malaria patients hospitalized from 1995 to 1998, 608 had severe malaria and 83 died. Acute renal failure, jaundice and respiratory distress were common in adults whereas children frequently had severe anaemia. Cerebral malaria occurred equally in adults and children but recovery from coma was quicker in children. Multiple complications caused high mortality in adults. PMID- 12886809 TI - Differences in automated depolarization patterns of Plasmodium falciparum and P. vivax malaria infections defined by the Cell-Dyn CD4000 haematology analyser. AB - Of 1014 samples submitted for full blood count analysis and malaria screening, 854 were designated malaria-negative by blood film microscopy, 79 were unequivocally identified as Plasmodium vivax and 81 as P. falciparum. All samples were additionally analysed with the Abbott Cell-Dyn CD4000 haematology instrument, and leucocyte differential plots of 90 degrees polarized vs. 90 degrees depolarized (NEU-EOS plot) and 90 degrees depolarized vs. 0 degree light (EOS I plot) scatter were specifically examined for abnormal depolarization patterns. Depolarization pattern types were correlated with microscopy (species) results, and these correlations were consolidated by polymerase chain reaction analysis. All 854 microscopically-designated malaria-negative samples showed a type 1 (normal) CD4000 depolarization pattern. Abnormal pattern types 2, 3a and 3b were entirely restricted to one of the two malaria categories. Plasmodium falciparum malaria showed two CD4000 pattern types only; a 'normal' type 1 pattern was seen in 36/75 (48%) cases and the remaining 39 cases were all abnormal pattern type 3a. In contrast, most (79/85) P. vivax malaria cases showed a distinctive clustered EOS I population (types 2 and 3b patterns) that was not seen with P. falciparum. Automated depolarization analysis provides an effective means of detecting malaria-associated haemozoin, and the patterns of intracellular haemozoin further appear to provide species differentiation between P. falciparum and P. vivax. PMID- 12886810 TI - Direct identification of Leishmania species in biopsies from patients with American tegumentary leishmaniasis. AB - Accurate identification of Leishmania species is important for monitoring clinical outcome, adequately targeting treatment, and evaluation of epidemiological risk in tegumentary leishmaniasis. This is especially the case in regions where several species coexist and for travel medicine where the geographical source of infection is not always obvious. Species identification presently depends on parasite isolation, which is not very sensitive and not necessarily representative of parasites actually present in human tissues. We evaluated a polymerase chain reaction (PCR) assay combining amplification of the gp63 genes and restriction fragment length polymorphism (RFLP) analysis (gp63 PCR RFLP) for direct Leishmania species-identification in tissues collected from Peruvian patients in 1999. By comparison with a kinetoplast DNA-based PCR, our PCR assay showed a detection sensitivity of 85%. Three species were encountered among patient samples, Leishmania (Viannia) braziliensis, L. (V.) peruviana and L. (V.) guyanensis, and their frequency and geographical distribution corresponded to earlier epidemiological studies of leishmaniasis in Peru. However, unexpected results raised questions about (i) the contribution of human migration to the emergence of new foci of given species, (ii) the pathogenicity of some species, and (iii) the frequency of mixed or hybrid infections. PMID- 12886811 TI - Influence of infection with non-filarial helminths on the specificity of serological assays for antifilarial immunoglobulin G4. AB - Serological assays based on the detection of immunoglobulin (Ig) G4 antibodies to crude filarial extracts are widely used for epidemiological and diagnostic purposes. We tested 195 samples collected in 1998 from an area of Brazil where filariasis is not endemic and 13 (6.7%) had levels of antifilarial IgG4 antibodies that were defined as positive. Both Strongyloides infection and the presence of Strongyloides antibody responses were associated with higher antifilarial antibody responses. None of the specimens had a positive response to the Brugia malayi recombinant antigen (Bm14). These data suggest that serodiagnostic assays based on the use of crude filarial antigens should be interpreted with caution because of the potential for cross-reactivity with Strongyloides. PMID- 12886812 TI - Prevalence and indicators of HIV and AIDS among adults admitted to medical and surgical wards in Blantyre, Malawi. AB - Despite high seroprevalence there are few recent studies of the effect of human immunodeficiency virus (HIV) on hospitals in sub-Saharan Africa. We examined 1226 consecutive patients admitted to medical and surgical wards in Blantyre, Malawi during two 2-week periods in October 1999 and January 2000: 70% of medical patients were HIV-positive and 45% had acquired immune deficiency syndrome (AIDS); 36% of surgical patients were HIV-positive and 8% had AIDS. Seroprevalence rose to a peak among 30-40 year olds; 91% of medical, 56% of surgical and 80% of all patients in this age group were HIV-positive. Seropositive women were younger than seropositive men (median age 29 vs. 35 years, P < 0.0001). Symptoms strongly indicative of HIV were history of shingles, chronic diarrhoea or fever or cough, history of tuberculosis (TB), weight loss and persistent itchy rash (adjusted odds ratios [AORs] all > 5). Clinical signs strongly indicative of HIV were oral hairy leukoplakia, shingles scar, Kaposi's sarcoma, oral thrush and hair loss (AORs all > 10). Of surgical patients with 'deep infections' (breast abscess, pyomyositis, osteomyelitis, septic arthritis and multiple abscesses), 52% were HIV-positive (OR compared with other surgical patients = 2.4). Severe bacterial infections, TB and AIDS caused 68% of deaths. HIV dominates adult medicine, is a major part of adult surgery, is the main cause of death in hospital and affects the economically active age group of the population. PMID- 12886813 TI - Virulence of variola viruses for suckling mice. AB - We report work done in 1971 to determine the quantitative virulence for suckling mice of 26 variola virus isolates from different countries and from cases of differing severity. Strains of recognized variola major and variola minor viruses differed up to 100-fold (expressed as the harmonic mean dose of inoculum which killed mice 2-4 d old, inoculated intracranially, in 5 d). Isolates from Indonesia and from East and West Africa gave intermediate values. Unlike tests on chick embryos, this test distinguished between African and Indonesian isolates. PMID- 12886814 TI - Case report: Schistosoma mansoni infection: failure of standard treatment with praziquantel in a returned traveller. AB - Heavy reliance upon praziquantel for the treatment of schistosomiasis raises concerns over potential development of drug resistance. We describe a British traveller who acquired Schistosoma mansoni infection in East Africa and in whom repeated standard 40 mg/kg doses of praziquantel failed to clear the infection despite no opportunity for reinfection. PMID- 12886815 TI - Associations between helminth infection and CD4+ T cell count, viral load and cytokine responses in HIV-1-infected Ugandan adults. AB - It has been proposed that helminth infection may exacerbate HIV progression by promoting activation of 'type 2' immune responses. To examine this hypothesis, we investigated helminth infection in a cohort of HIV-1-seropositive adults in Entebbe, Uganda, during November 1999 to January 2000. Individuals with helminths were treated. At enroLlment, after 5 weeks and after 4 months, CD4+ and CD8+ T cell counts and viral load were measured. Cytokine responses (interferon [IFN] gamma, interleukin [IL]-2, IL-4 and IL-5) to Schistosoma mansoni adult worm antigen (SWA), Mycobacterium tuberculosis culture filtrate proteins (CFPs) and phytohaemagglutinin (PHA) were measured in a whole blood assay. At baseline, CD4+ T cell counts and CD4+: CD8+ ratios were higher in individuals with helminths than in those without (median CD4+ T cell counts 467/microL and 268/microL, respectively, P = 0.005). Viral load was lower among those with helminths but this was not statistically significant. During follow-up, CD4+ T cell counts and cytokine responses to PHA fell among individuals without helminths. Among those treated for helminths, CD4+ counts remained stable. Viral loads showed a transient increase at 5 weeks, which was more marked among those treated for helminths, but the levels at 4 months were similar to baseline in both groups. Among those with schistosomiasis, IFN-gamma and IL-2 responses to CFP, and IL-2 and IL-4 responses to PHA declined but there was a sustained increase in cytokine responses to SWA following treatment. These data do not support the hypothesis that helminth infection exacerbates HIV infection. The possibility that chronic helminth infection may suppress HIV replication and that effects on HIV replication may vary during helminth infection and treatment should be considered. PMID- 12886816 TI - Effects of multimicronutrient supplementation on helminth reinfection: a randomized, controlled trial in Kenyan schoolchildren. AB - A randomized, placebo-controlled, double-blind, two-by-two factorial trial was carried out among 977 schoolchildren from 19 primary schools in Nyanza Province, Kenya from February 1995 to February 1996. The interventions were multimicronutrient supplementation (vitamin A, 1000 micrograms; vitamin B1, 1.4 mg; vitamin B2, 1.6 mg; vitamin B6, 1.7 mg; vitamin B12, 2.0 micrograms; folate, 150 micrograms; niacin, 16 mg; vitamin C, 50 mg; vitamin D, 5 micrograms; vitamin E, 8 mg; iron, 18 mg; zinc, 20 mg; copper, 2.0 mg; iodine, 150 micrograms; selenium, 40 micrograms) and multihelminth chemotherapy (albendazole 600 mg in a single dose and/or praziquantel 40 mg/kg in a single dose). This paper reports the effects of the supplementation given on all school days on reinfection with hookworm, Ascaris lumbricoides, Trichuris trichiura and Schistosoma mansoni after 11 months. Baseline prevalence and geometric mean intensity for hookworm, A. lumbricoides, T. trichiura and S. mansoni in all children investigated were 54.7%, 13.8%, 45.6% and 70.0%, respectively and 8.6, 2.7, 5.9 and 19.4 eggs per gram (epg), respectively. Children received a mean of 2.3 multimicronutrient/placebo tablets per school week, giving a compliance rate of 46%. Children given multimicronutrients had a slightly, but significantly, lower intensity of S. mansoni reinfection compared with children given placebo (5.5 epg vs. 7.7 epg, P = 0.047). Multiple linear regression analyses controlling for baseline infection status confirmed this, as children who received micronutrients were reinfected with S. mansoni at only 69% of the intensity of those who received placebo. Multiple logistic regression analyses revealed that micronutrient supplementation was associated with a lower S. mansoni reinfection rate (odds ratio = 0.7) although this was only of borderline significance (P = 0.090). There were no significant differences in reinfection rates or intensities of hookworm, A. lumbricoides and T. trichiura. The effect on S. mansoni infection intensity is particularly interesting given the low compliance, suggesting that full micronutrient supplementation might have a role to play in S. mansoni control programmes. PMID- 12886817 TI - Increased nitric oxide production in acute diarrhoea is associated with abnormal gut permeability, hypokalaemia and malnutrition in tropical Australian aboriginal children. AB - Australian Aboriginal children hospitalized with diarrhoeal disease have severe manifestations with acidosis, hypokalaemia, osmotic diarrhoea and abnormal small bowel permeability. Nitric oxide (NO) production is increased in diarrhoeal disease, but its relationship to mucosal function and diarrhoeal complications is not known. We examined the relationship between NO production and complications of acute diarrhoea in Aboriginal and non-Aboriginal children between February 1998 and February 2000. We enrolled 318 children admitted to Royal Darwin Hospital into one of three groups: acute diarrhoea, non-diarrhoeal controls with no inflammatory illness, and non-diarrhoeal controls with inflammatory illness. Nitric oxide production was measured by urine nitrate-creatinine (NOx/Cr) excretion on a low nitrate diet. Small bowel intestinal permeability was measured by the lactulose-rhamnose (L/R) ratio on a timed blood specimen. The NOx/Cr ratios were markedly elevated in Aboriginal diarrhoeal cases (geometric mean [GM] = 1.23, 95% confidence interval [95% CI] 1.07-1.44), lowest in non-Aboriginal non inflammatory controls (GM = 0.13, 95% CI 0.10-0.16) and intermediate in all other groups (GM = 0.35, 95% CI 0.28-0.43). Convalescent levels (day 5) in the Aboriginal diarrhoeal group (GM = 1.02, 95% CI 0.82-1.28) were slower to fall than L/R ratios. Multivariate analysis in the diarrhoeal group indicated that high NO production was associated with abnormal permeability, hypokalaemia and malnutrition, but not with the severity of diarrhoea, acidosis or osmotic diarrhoea. We concluded that increased NO production may contribute to impaired mucosal barrier function and hypokalaemia in acute gastroenteritis, which may be the cost of the known gut-protective and antimicrobial effects mediated by NO in acute intestinal inflammation. PMID- 12886818 TI - Haptoglobin genotypes are not associated with resistance to severe malaria in The Gambia. PMID- 12886819 TI - Re-treatment of bednets in Tanzania. PMID- 12886821 TI - [Chronic hepatopathies: from cryptogenetics to multiple factors]. PMID- 12886820 TI - Mechanisms involved in myocardial necrosis and pulmonary oedema after Tityus serrulatus scorpion envenomation. PMID- 12886822 TI - [Expression of Herpesviridae diseases: epidemiological, immunopathological, and clinical reflections]. PMID- 12886823 TI - Urban air pollution at the crossroads of the allergic pandemic. AB - In these past decades an important increase in the prevalence of allergic respiratory diseases has been documented in most countries of the world with large differences being reported within different areas, particularly in industrialized countries. Persistent environmental exposure to particulate air pollution from motor vehicles has been suggested to be an important factor contributing to the observed increased prevalence of allergic diseases. Data from various investigators in different parts of the world have shown an important association between environmental levels of motor vehicle exhaust emissions and increased symptoms of asthma and rhinitis. In addition, recent human and animal laboratory-based studies have shown that particulate toxic pollutants, and especially diesel exhaust particles, can enhance allergic inflammation and induce the development of allergic immune responses. This article reviews the current state of knowledge on the role of diesel exhaust particles in the susceptibility to allergy. It scrutinizes the epidemiological evidence that supports the causative link between particulate air pollution from motor vehicles and the increasing prevalence in allergic conditions and the immunologic mechanisms by which diesel exhaust particles enhance the susceptibility to allergy. PMID- 12886824 TI - [Curing smoking: a priority health objective]. AB - Cigarette smoking is the number one avoidable killer in the industrial world. Yet doctors frequently do not treat or prevent the disease as they should. Education and training are lacking. In the present article an overview of these and other smoking-related problems and, at the same time, a description of the practical management of an outpatient clinic for smoking cessation, the epidemiology of smoking, the related risks and mortality, as well as the advantages of quitting are presented. The toxicology and pharmacology are discussed, in relation to addiction and craving. The article also includes the diagnosis of nicotine dependence and nicotine replacement as well as bupropione and acupuncture treatments. The gain in body weight, depression and relapse are also dealt with. The various and personally tailored therapeutic strategies used in a service operating in a setting of Internal Medicine are presented and discussed. PMID- 12886825 TI - Clinical features of cytomegalic disease in immunocompetent subjects: a 1990-2000 survey. AB - All cases of Cytomegalvirus-related disease in previously healthy immunocompetent subjects admitted to the University Hospital of Catania between 1990 and 2000 were reviewed. Eighty-one immunocompetent subjects were discharged with a diagnosis of acute cytomegalic disease. Nevertheless, only in 26 cases was the diagnosis confirmed by the determination of the specific immunoglobulin M, viremia or antigenemia. Fifteen subjects presented with a mononucleosis-like syndrome. Eleven subjects had a more severe form of the disease with organ involvement: 3 hepatitis, 3 interstitial pneumonia, 2 aseptic meningitis, 2 ulcerative colitis and 1 peripheral vasculitis. All patients recovered. Two out of 11 severe cases were treated with ganciclovir and 7 with steroids; the remainder received only supportive treatment. Large-scale surveys are required to assess the real impact of Cytomegalovirus disease in immunocompetent subjects and to elaborate guidelines for the management of severe cases. PMID- 12886826 TI - [The use of parenteral clodronate in elderly women with postmenopausal osteoporosis: compliance, effects on bone mineral density and on bone turnover]. AB - The aim of the study was to evaluate parenteral clodronate (CLD) compliance in patients with postmenopausal osteoporosis and intolerance to aminobisphosphonates. Moreover, we have also assessed the effects of CLD on bone mineral density (BMD) and bone turnover. Eighty-four consecutive postmenopausal women with osteoporosis (range 62-74 years) were enrolled and randomly allocated to three groups: group A included 26 women who received CLD i.v., 300 mg/2 weeks and oral supplemental calcium carbonate (500 mg x 2/day) and vitamin D3 (400 IU x 2/day); group B included 28 women who received CLD i.m., 100 mg/week, and the same dose of calcium and vitamin D3 administered to group A; group C, the control group, included 30 women receiving only calcium and vitamin D3 at the same doses as the other two groups. The lumbar spine (L1-L4) and femoral neck (FN) BMD were measured by dual energy X-ray absorbiometry at time 0 (T0) and after 6 (T6), 12 (T12), 18 (T18) and 24 (T24) months. At the same time, the serum bone specific alkaline phosphatase and amino-terminal telopeptide of type I collagen normalized by creatinine (NTx/cr) were determined at T0, T6, T12, T18, and T24. Eighty (95.2%) women completed the study, 24 in group A, 27 in group B and 29 in group C. In groups A and B, after 6 months of treatment we found a significantly greater (p < 0.05) increase in the L1-L4 BMD with respect to group C. After 12 months of therapy, in group A the L1-L4 BMD (1.8 +/- 0.5%) was significantly higher (p < 0.05) than that in group B (0.9 +/- 0.3%). At the end of the study, in groups A (1.2 +/- 0.5%) and B (1.1 +/- 0.4%) the percentage increase in the FN BMD was significantly greater (p < 0.05) than in group C (0.6 +/- 0.5%). After 24 months of therapy, there was no difference in the FN BMD between groups A and B. Since the sixth month, both the bone specific alkaline phosphatase and NTx/cr were found to be more markedly and significantly decreased (p < 0.05) in groups A and B with respect to group C. After 18 months, in group A (NTx/cr -16.7 +/- 0.8%) we observed a significantly reduced (p < 0.05) bone resorption with respect to group B (NTx/cr -11.0 +/- 0.5%). In group B, only 3 patients (11.2%) referred pain at the site of drug administration. Our data demonstrate that compliance to parenteral CLD is satisfactory and that this drug reduces bone turnover, increases the L1-L4 BMD and decreases the FN BMD loss. Parenteral CLD administration can represent an effective alternative treatment for postmenopausal women with osteoporosis, especially those who do not tolerate oral aminobisphosphonates. PMID- 12886827 TI - [Multifactorial hepatopathy in a patient with biliopancreatic diversion]. AB - The Scopinaro surgical technique for obesity consists of a partial distal gastrectomy with biliopancreatic diversion and cholecystectomy. It is an effective procedure in the control of the body weight but disrupts the physiological gut-liver axis. We report the case of a patient who developed liver cirrhosis with a multifactorial pathogenesis following antiobesity surgery according to Scopinaro and discuss the pathogenesis of the liver damage on the basis of our present understanding of alcoholic and nonalcoholic steatohepatitis. A 41-year-old male patient presented with ascites due to cryptogenic liver cirrhosis. Owing to morbid obesity, he had undergone antiobesity surgery according to Scopinaro when he was 25 years old. The procedure was effective and the patient lost about 40 kg of weight but manifested chronic diarrhea in the postoperative course. During the following 15 years, the patient continued to assume 100 g alcohol/day and did not turn up for clinical evaluation. Signs and symptoms of liver failure appeared in May 2000. After surgical correction of biliopancreatic diversion and abstinence from alcohol the hepatocellular function partially recovered and the patient regained his subjective complete well-being. The multifactorial pathogenesis of the liver disease observed in our patient included obesity, antrectomy, heavy alcohol consumption, bacterial overgrowth, and protein malnutrition. PMID- 12886828 TI - [Spontaneous splenic rupture as complication of infective mononucleosis: a clinical case]. AB - Spontaneous splenic rupture is a very rare complication of infectious mononucleosis, already described by other authors. In this case report we show the findings occurring in a young man who presented with sudden left thoracic pain and dyspnea. The initial work-up was oriented towards a suspected diagnosis of left spontaneous pneumothorax. Initially, little attention was paid to the clinical history that was characterized by flu-like symptoms during the previous 2 weeks; this, along with a rushed physical examination, could have had dramatic consequences, due to the unavoidable diagnostic delay. A thorough clinical evaluation by Internists, coupled with the essential ultrasonographic diagnostic support, allowed an earlier diagnosis followed by definitive cure, i.e. splenectomy. PMID- 12886829 TI - [Autoimmune hemolytic anemia presenting during treatment of chronic hepatitis C with interferon alpha]. AB - Autoimmune hemolytic anemia has been rarely described in patients with chronic hepatitis C during interferon-alpha therapy. We report a case of Coombs-positive autoimmune hemolytic anemia, occurring 9 months after the onset of monotherapy with interferon-alpha-2b (3 million units thrice in week) for chronic hepatitis C. Interferon withdrawal and prednisone treatment induced a complete remission of hematological findings after 3 months. During a follow-up period of 30 months after cessation of interferon, hemoglobin levels stabilized about 16 g/dL, and the indirect Coombs test turned negative, while the direct Coombs test remained weakly positive; besides, serum alanine aminotransferase values were normal and hepatitis C virus-RNA remained undetectable. In our patient, clinical and laboratory findings suggested a correlation between autoimmune side effects of interferon-alpha and autoimmune hemolytic anemia. PMID- 12886830 TI - [Postgraduate teaching of internal medicine in the United States]. AB - The structure and organization of post-graduate training in Internal Medicine in the United States is described as compared to the Italian situation. The main differences are highlighted and discussed, with particular emphasis on the central role that education in Internal Medicine has in the United States for medical subspecialties as well. PMID- 12886831 TI - Scoring systems for drug-induced hepatotoxicity. PMID- 12886834 TI - Meeting the needs of mothers with disabilities. PMID- 12886835 TI - Beating disability, embracing motherhood. PMID- 12886836 TI - Second-class service: a visually impaired mother's story. PMID- 12886838 TI - A guide to maternity and parental rights. PMID- 12886839 TI - Pushing the boundaries: independence in the NHS. PMID- 12886840 TI - Pension options for health professionals. PMID- 12886841 TI - What's it like to work in Siberia? PMID- 12886842 TI - Type 2 diabetes and its impact on pregnancy. PMID- 12886843 TI - The epidural: a barrier to 'natural' birth? PMID- 12886844 TI - Alternidurals. PMID- 12886845 TI - The NHS and leadership. PMID- 12886846 TI - Case reports: here or to go? PMID- 12886847 TI - Vitreous haemorrhage in the diabetic eye. AB - Vitreous haemorrhage is a serious sign indicating significant intraocular pathology in the diabetic eye and warranting prompt referral to an ophthalmologist for further assessment and management. PMID- 12886848 TI - Epidemiology of diabetic retinopathy. AB - Diabetic retinopathy is associated with a number of systemic risk factors, namely hyperglycaemia, elevated blood pressure and dyslipidaemia. Patients with diabetes should be vigorously treated for these modifiable risk factors to prevent the development and progression of diabetic retinopathy. PMID- 12886849 TI - Diabetic retinopathy: medical management and assessment. AB - Diabetic retinopathy is a common and potentially devastating complication. This review discusses the practical assessment and management of retinopathy, focusing on the treatment of concomitant systemic disorders that influence the development and progression of this complication. PMID- 12886850 TI - Principles of management in oral cancer. AB - Squamous cell carcinoma is the most common oral malignancy, with a relatively poor prognosis. Treatment of oral cancer has a major impact on afflicted patients because it affects speech, swallowing and mastication. Surgery is the main treatment of oral cancer, as a single modality or combined with radiotherapy. Vigilance is vital for early diagnosis and better overall prognosis. PMID- 12886851 TI - Laparoscopic surgery in urology: nephrectomy and prostatectomy. AB - Urological laparoscopic techniques have received great attention in the past decade. With the development of improved laparoscopic instrumentation, approaches to kidney and prostatic diseases have been successfully performed. Laparoscopic nephrectomy and prostatectomy are discussed. Awareness of these evolving technologies is critical for all surgeons with an interest in laparoscopy. PMID- 12886852 TI - The working day in medicine: lessons from the air. AB - This article discusses whether it would be wrong to explain a mistake involving medical responsibility on the basis of an opinion that the professional was not in the optimum physical or mental state at the moment the mistake took place. PMID- 12886853 TI - HIV/AIDS and the eye. PMID- 12886854 TI - Cardiopulmonary resuscitation: attitudes and perceptions of junior doctors. AB - Many junior doctors feel cardiac arrests are unsatisfactorily managed and experience high levels of stress during the procedure, often feeling they are inadequately trained for the task. Juniors also regard 'do not resuscitate' orders as decisions for teams, not individuals. PMID- 12886855 TI - The controversy of breast cancer screening. PMID- 12886856 TI - Acute interstitial nephritis and COX-2 inhibition. PMID- 12886857 TI - Strangulated hernias secondary to generalized peritonitis: an old lesson relearnt. PMID- 12886858 TI - Acute pancreatitis presenting as a case of splenic rupture. PMID- 12886859 TI - Chronic appendicitis diagnosed incidentally by colonoscopy. PMID- 12886860 TI - Small bowel perforation caused by an ingested foreign body masquerading as acute appendicitis. PMID- 12886861 TI - Laryngeal mask or tube for tonsillectomy? PMID- 12886862 TI - Code of ethics for nurses with interpretive statements. PMID- 12886863 TI - Alabama State Nurse Association: clearing the myths and misconceptions . PMID- 12886864 TI - Prophylactic surgery in hereditary breast/ovarian cancer syndrome. AB - The hereditary breast/ovarian cancer syndrome is responsible for approximately 5% of all breast cancers and 10% of all ovarian cancers. Although this accounts for a small portion of these diseases, much attention has been focused on this syndrome because of the abundance of research in this area. The majority of the hereditary breast/ovarian syndrome can be attributed to germ-line mutations in the BRCA1 and BRCA2 genes. Reliable screening techniques for these mutations have been developed and are readily available in clinical practice. For patients who are thought to have the hereditary breast/ovarian cancer syndrome based on family history or genetic testing, options exist for either intensive screening or prophylactic surgery. This review will discuss the mechanisms by which mutations in the BRCA genes lead to the development of cancer, the limitations of currently available screening techniques, and the efficacy of prophylactic surgery. In general, prophylactic oophorectomy can be performed laparoscopically as an outpatient procedure, carrying as its main drawback the associated consequence of surgical menopause. Prophylactic mastectomy is quite effective in reducing the risk of breast cancer but is a more extensive surgical procedure and results in disfigurement. For any given patient, the best estimates of individual risk of breast or ovarian cancer should be weighed against the benefits of prophylactic surgery and the patient's personal wishes. PMID- 12886865 TI - The benefits of achieving stable disease in advanced lung cancer. AB - The cytostatic, molecular-targeted therapies becoming available for lung cancer and other human solid tumors are more likely to result in stable disease than to produce tumor regression. In the setting of advanced lung cancer, stable disease provides significant benefit to the patient. However, in the context of clinical trials, stable disease is vaguely defined, difficult to measure, and may represent a heterogeneous patient population. The inclusion of alternative trial end points such as symptom improvement and biologic activity may help to identify patients who have achieved clinically relevant stable disease. The epidermal growth factor receptor-tyrosine kinase inhibitor gefitinib (Iressa) has been shown to produce partial responses and stable disease in patients with advanced lung cancer who have previously received treatment with standard chemotherapies. In the monotherapy trials of gefitinib, stable disease was correlated with improvements in disease-related symptoms and quality of life--the most meaningful end points for the patient with advanced lung cancer. Thus, with the introduction of new molecular-targeted agents, stable disease with clinical benefit should become an important goal of anticancer therapy. PMID- 12886866 TI - Multidisciplinary management of paragangliomas of the head and neck, Part 1. AB - Paragangliomas most commonly occur in the carotid body, jugulotympanic area, and vagus nerve but have also been reported in other areas of the head and neck. These tumors are highly vascular and characteristically have early blood vessel and neural involvement, making their treatment particularly challenging. Surgery has traditionally been the preferred method of treatment, especially in light of recent advances in technique. However, compared to radiation therapy, it can result in a higher incidence of cranial nerve dysfunction. Radiation therapy has the advantage of avoiding the increased morbidity of surgery while offering an equal possibility of cure. Part 1 of this two-part article focuses on techniques for diagnosing paraganglioma and the indications for and use of surgery as primary treatment. The complications commonly associated with surgery are reviewed, and strategies for rehabilitation of affected patients are presented. PMID- 12886867 TI - Sentinel lymph node biopsy in a young child with thick cutaneous melanoma. AB - Head and neck melanoma is a rare and aggressive childhood malignancy. Surgery remains the primary treatment, with lymphatic involvement determined by neck dissection. In the adult population, sentinel lymph node biopsy has emerged as a less morbid yet accurate method of staging regional lymph nodes. This innovative technique can also be used in the pediatric population. PMID- 12886868 TI - Population-based maximum tolerated dose of irinotecan and carboplatin. AB - A novel schema of intrapatient dose escalation was applied to determine a population-based maximum tolerated dose (pMTD) for irinotecan (CPT-11, Camptosar) and carboplatin (Paraplatin) in a phase I trial. A total of 74 patients with advanced solid tumors were enrolled with the following characteristics: men/women, 46/28; median age, 61 years; 51 patients with and 23 patients without prior chemotherapy; performance status of 0-1 (93%) and 2 (7%). Patients were started at dose level 1 with irinotecan at 200 mg/m2, and carboplatin at an area under the concentration-time curve (AUC) of 5 mg/mL x min, administered every 21 days. Depending on degree of toxicity observed, the dose for each patient in each subsequent cycle was determined according to a predetermined schema of dose levels. Individual maximum tolerated dose (iMTD) was determined for each patient. The pMTD was defined as the highest dose level for which the incidence of dose limiting toxicity occurred in less than 33% of the patient population. The most common dose-limiting toxicity included neutropenia (58%), thrombocytopenia (15%), diarrhea (8%), and nausea/emesis (7%). The iMTD ranged from dose level-3 (irinotecan at 100 mg/m2 and carboplatin at an AUC of 4) to dose level 5 (irinotecan at 350 mg/m2 and carboplatin at AUC 6). The pMTD was determined to be dose level-1 and 1 for previously chemotherapy-treated and--untreated patients, respectively. Fifty-nine patients were assessable for response. Of note, a response rate of 40% was observed in 15 patients with relapsed small-cell lung cancer previously treated with platinum-based therapy. We recommend dose level 1 of irinotecan (200 mg/m2) and carboplatin (AUC 5) for chemotherapynaive patients, and dose level-1 of irinotecan (150 mg/m2) and carboplatin (AUC 5) for chemotherapy-treated patients in phase II trials. PMID- 12886869 TI - Dose-dense therapy with a novel irinotecan regimen for small-cell lung cancer. AB - Irinotecan (CPT-11, Camptosar) has significant activity in small-cell lung cancer. In addition, preclinical models have demonstrated synergy between irinotecan and two standard front-line drugs for small-cell lung cancer: cisplatin and etoposide; phase III data also show significant survival benefit for irinotecan/cisplatin compared to standard treatment. These data suggest a potential clinical advantage to combining etoposide, cisplatin, and irinotecan as first-line therapy for small-cell lung cancer. The purpose of this phase I study was to establish if alternating weekly therapy with irinotecan/cisplatin and etoposide/cisplatin was tolerated and to determine the maximum tolerated dose of these agents in patients with small-cell lung cancer. Patients generally tolerated the weekly alternating chemotherapy with irinotecan/cisplatin and etoposide/cisplatin well, and this combination possessed significant antitumor activity in small-cell lung cancer. PMID- 12886870 TI - Irinotecan, cisplatin/carboplatin, and COX-2 inhibition in small-cell lung cancer. AB - Recent findings indicate significant prolongation of survival and time to disease progression with irinotecan (CPT-11, Camptosar)/cisplatin vs etoposide/cisplatin in extensive-stage small-cell lung cancer, and a larger-scale phase III trial has been planned to provide more definitive data on the benefits of the irinotecan/cisplatin combination in this setting. Early-phase studies indicate that the activity of carboplatin (Paraplatin) in small-cell lung cancer is comparable to that of cisplatin, and that combining irinotecan on a day 1 and 8 schedule with split-dose carboplatin is feasible. Inhibition of the cyclooxygenase-2 (COX-2) enzyme, which is active in tumorigenesis, may augment efficacy and reduce toxicity of platinum/irinotecan combinations. A phase II trial has been designed to compare irinotecan/carboplatin and irinotecan/cisplatin combinations with or without the COX-2 inhibitor celecoxib (Celebrex) in patients with extensive-stage small-cell lung cancer. Results of these trials will help define the roles of platinum/irinotecan combinations and COX-2 inhibition in treatment for small-cell lung cancer. PMID- 12886871 TI - Irinotecan and carboplatin in metastatic or recurrent non-small-cell lung cancer. AB - The outcome of patients with metastatic lung cancer is poor, with a 1-year survival rate of approximately 35%. We are evaluating the combination of irinotecan (CPT-11, Camptosar) and carboplatin (Paraplatin) in patients with stage IIIB and IV non-small-cell lung cancer. Eligible patients include those with histologic or cytologic diagnosis of non-small-cell lung cancer; no previous chemotherapy for metastatic lung cancer; and Eastern Cooperative Oncology Group performance status 0 or 1. The first five patients received irinotecan at 250 mg/m2 over 90 minutes followed by carboplatin at an area under the concentration time curve (AUC) of 5 over 1 hour. Subsequently, the dose of irinotecan was reduced to 200 mg/m2 in view of febrile neutropenia in one of five patients. Chemotherapy cycles are repeated every 21 days. Patients are reevaluated every two cycles. Of a planned 37 patients, 14 have been enrolled and 9 are evaluable for toxicity and response at the time of this report. Thirty-two cycles have been administered, with seven 1-week delays and two dose reductions. To date there have been two partial responses and five patients with stable disease; two patients developed progressive disease on therapy. One patient had neutropenic fever; other toxicities included mild pancreatitis (n = 1) and diverticulitis (n = 1). The regimen of irinotecan and carboplatin administered once every 3 weeks demonstrates early evidence of activity, and is tolerable and convenient. The main toxicity is hematologic. This study is ongoing and actively accruing patients. PMID- 12886872 TI - The global role of irinotecan in the treatment of lung cancer: 2003 update. AB - Lung cancer is the most lethal malignancy in the United States. In light of its natural history, new agents with improved systemic activity are needed. Irinotecan (CPT-11, Camptosar) is a promising agent in the treatment of advanced non-small-cell and small-cell lung cancer. In subgroup analysis of a Japanese phase III trial, irinotecan or irinotecan/cisplatin demonstrated a significant survival advantage compared with standard vindesine (Eldisine)/cisplatin in advanced non-small-cell lung cancer. Similar North American phase III trials focusing on irinotecan's role in non-small-cell lung cancer are planned. Ongoing trials have also been inaugurated to corroborate the significant survival advantage reported by a Japanese phase III trial of irinotecan/cisplatin vs standard etoposide/cisplatin in extensive small-cell lung cancer. Current and planned trials in patients with non-small-cell lung cancer will investigate treatment using irinotecan in combination with gemcitabine (Gemzar), the taxanes, and other new agents, as well as with thoracic radiotherapy. Moreover, trials in small-cell lung cancer are investigating the utility of irinotecan in combination with a platinum agent when incorporated in chemoradiotherapy regimens. It is hoped that data from these and other studies will help investigators to more clearly delineate a role for irinotecan in the management of lung cancer. PMID- 12886873 TI - Modelling autocatalytic networks with artificial microbiology. AB - Cells can usefully be equated to autocatalytic networks that increase in mass and then divide. To begin to model relationships between autocatalytic networks and cell division, we have written a program of artificial chemistry that simulates a cell fed by monomers. These monomers are symbols that can be assembled into linear (non-branched) polymers to give different lengths. A reaction is catalysed by a particular polymer or 'enzyme' that may itself be a reactant of that reaction (autocatalysis). These reactions are only studied within the confines of the 'cell' or 'reaction chamber'. There is a flux of material through the cell and eventually the mass of polymers reaches a threshold at which we analyse the cell. Our results indicate a similarity between the connectivity of the reaction network and that of real metabolic networks. Developing the model will entail attributing increased probabilities of reactions to polymers that are colocalised to evaluate the consequences of the dynamics of large assemblies of diverse molecules (hyperstructures) and of cell division. PMID- 12886874 TI - Dynamin recruitment by clathrin coats: a physical step? AB - Recent structural findings have shown that dynamin, a cytosol protein playing a key-role in clathrin-mediated endocytosis, inserts partly within the lipid bilayer and tends to self-assemble around lipid tubules. Taking into account these observations, we make the hypothesis that individual membrane-inserted dynamins imprint a local cylindrical curvature to the membrane. This imprint may give rise to long-range mechanical forces mediated by the elasticity of the membrane. Calculating the resulting many-body interaction between a collection of inserted dynamins and a membrane bud, we find a regime in which the dynamins are elastically recruited by the bud to form a collar around its neck, which is reminiscent of the actual process preempting vesicle scission. This physical mechanism might therefore be implied in the recruitment of dynamins by clathrin coats. PMID- 12886875 TI - Adaptive branching in evolution and epigenesis. AB - We describe one of the simplest models that exhibit an adaptive branching behaviour. It is analysed both experimentally and formally, and its successive bifurcations provide a good model of what R. Thom called 'generalized catastrophes'. Two theorems on the stochastic adaptivity of the algorithm to very general shapes of target are given. The model further displays the phenomenon of abortive branching: each macroscopic branching appears after a burst of microscopic branchings that stop growing after a very short time. The mathematical analysis of the model explains why and how this behaviour occurs. Possible applications of these models to Evolution (natural and artificial) and Epigenesis are briefly mentioned, and a higher dimensional version is applied to growing a tree in a space of shapes in the context of a database of medical images. PMID- 12886876 TI - Bio-array images processing and genetic networks modelling. AB - The new tools available for gene expression studies are essentially the bio-array methods using a large variety of physical detectors (isotopes, fluorescent markers, ultrasounds...). Here we present first rapidly an image-processing method independent of the detector type, dealing with the noise and with the peaks overlapping, the peaks revealing the detector activity (isotopic in the presented example), correlated with the gene expression. After this primary step of bio-array image processing, we can extract information about causal influence (activation or inhibition) a gene can exert on other genes, leading to clusters of genes co-expression in which we extract an interaction matrix M and an associated interaction graph G explaining the genetic regulatory dynamics correlated to the studied tissue function. We give two examples of such interaction matrices and graphs (the flowering genetic regulatory network of Arabidopsis thaliana and the lytic/lysogenic operon of the phage Mu) and after some theoretical rigorous results recently obtained concerning the asymptotic states generated by the genetic networks having a given interaction matrix and reciprocally concerning the minimal (in the sense of having a minimal number of non-zero coefficients) matrices having given stationary stable states. PMID- 12886877 TI - Dynamic simulation of pollutant effects on the threonine pathway in Escherichia coli. AB - The enzymatic activities of threonine pathway in Escherichia coli are sensitive to pollutants such as cadmium, copper and mercury, which, even at low concentration, can substantially decrease or even block the pathway at several steps. Our aim was to investigate the complex effects on a metabolic pathway of such general enzyme inhibitors with several sites of action, using a previously developed computer simulation of the pathway. For this purpose, the inhibition parameters were experimentally determined and incorporated in the model. The calculation of the flux control coefficient distribution between the five steps of the threonine pathway showed that control remains shared between the three first steps under most inhibition conditions. Response coefficient analysis shows that the inhibition of aspartate semialdehyde dehydrogenase is quantitatively dominant in most circumstances. PMID- 12886878 TI - Metabolic analysis in drug design. AB - Biotechnology is often presented as if progress in the past two decades represented a major success, but the reality is quite different. For example, ten major classes of antibiotics were discovered between 1935 and 1963, but after 1963 there has been just one, the oxazolidones. To illustrate the possibilities of doing better by taking account of the real behaviour of metabolic systems, we can examine how one might modify the activity of an enzyme in the cell (for example by genetic manipulation, or by the action of an inhibitor, etc.) to satisfy a technological aim. For example, if the objective is to eliminate a pest, one might suppose that the effect of an inhibitor could be to depress an essential flux to a level insufficient for life, or to raise the concentration of an intermediate to a toxic level. The former may seem the more obvious, but the latter is easier to achieve in practice, and there are some excellent examples of industrial products that work in that way, such as the herbicide 'Roundup' and antimalarials of the quinine class. A study of glycolysis in the parasite Trypanosoma brucei (which causes African sleeping sickness) indicates that for this approach to work the selected target enzyme must have a substrate with a concentration that is not limited by stoichiometric constraints. That is not necessarily easy to find in a complicated system, and typically needs the metabolic network to be analysed in the computer. PMID- 12886879 TI - Integrative biology: linking levels of organization. AB - Biological systems are composed of different levels of organization. Usually, one considers the atomic, molecular, cellular, individual, population, community and ecosystem levels. These levels of organization also correspond to different levels of observation of the system, from microscopic to macroscopic, i.e., to different time and space scales. The more microscopic the level is, the faster the time scale and the smaller the space scale are. The dynamics of the complete system is the result of the coupled dynamical processes that take place in each of its levels of organization at different time scales. Variables aggregation methods take advantage of these different time scales to reduce the dimension of mathematical models such as a system of ordinary differential equations. We are going to study the dynamics of a system which is hierarchically organized in the sense that it is composed of groups of elements that can be themselves divided into further smaller sub-groups and so on. The hierarchical structure of the system results from the fact that the intra-group interactions are assumed to be larger than inter-group ones. We present aggregation methods that allow one to build a reduced model that governs a few global variables at the slow time scale. PMID- 12886880 TI - Quality treat. PMID- 12886881 TI - All change. PMID- 12886882 TI - The hard cell. PMID- 12886883 TI - Shared services. Pay on display. AB - A year late, the electronic staff records project is now in the hands of the Department of Health. Pilots are underway, but it is unclear when every NHS payroll will be processed by the system. Progress has been slowed by the wide variety in organisations' structures and systems. PMID- 12886885 TI - Diabetes. Up--and downs. AB - A study examined different models of clinic practice for young people with diabetes, focusing particularly on the transfer to adult services. Young people favoured 'age-banded' clinics staffed by diabetes team members from both adult and paediatric services. Young people interviewed before transferring to adult care reported that they lacked information about their new clinic. PMID- 12886886 TI - Law. Ready, willing and liable. PMID- 12886887 TI - HSJ people. One for all. PMID- 12886888 TI - Imaging tutorial: differential diagnosis of bright lesions on diffusion-weighted MR images. AB - High sensitivity (94%) and specificity (100%) have been reported in the diagnosis of acute cerebral infarction with diffusion-weighted magnetic resonance (MR) imaging. However, high signal intensity on diffusion-weighted MR images and low apparent diffusion coefficient values (similar to the findings in acute cerebral infarction) were reported in such diverse conditions as hemorrhage, abscess, lymphoma, and even Creutzfeldt-Jakob disease. The differential diagnosis of these conditions (eg, acute ischemic infarction and acute cerebral hemorrhage) is critical for the determination of appropriate treatment. The authors present a systematic review of bright lesions on diffusion-weighted MR images and their differential diagnosis, with emphasis on the practical and clinical approaches of differential diagnosis. PMID- 12886889 TI - Real-time four-dimensional imaging of the heart with multi-detector row CT. AB - An interactive four-dimensional (4D) visualizing system for the heart was developed by the authors. The system realizes high-resolution three-dimensional (3D) imaging with temporal resolution in a beating heart by using eight or more data sets reconstructed from multi-detector row computed tomography (MDCT) with a retrospective electrocardiograph-gated reconstruction algorithm. The motion of heart walls, papillary muscles, septa, and valves can now be observed in 4D multiplanar reformations (MPRs), as with sonography, while coronary arteries, coronary sinuses, and cardiac veins can be analyzed during the optimal phase in 4D volume-rendering images, as with angiography. All parameters such as window width, window level, field of view, panning, tilt, thresholds, opacity, color, and segmentation function are completely interactive in 4D imaging. Two longitudinal views and one latitudinal view of a heart can be simultaneously visualized in the three relative 4D MPR views. These newly developed capabilities in viewing both 3D volume and temporal resolution data, functional data, and even multiphase data with registration add considerable diagnostic potential. The advent of this real-time 4D visualizing system has enhanced the capabilities of MDCT. PMID- 12886890 TI - Evaluation of cardiac valvular disease with MR imaging: qualitative and quantitative techniques. AB - Magnetic resonance (MR) imaging is almost never performed as the initial imaging test in cardiac valvular disease; that role is dominated by echocardiography. Nevertheless, MR imaging has much to offer in selected patients. Quantitative information regarding the severity of regurgitant or stenotic lesions can be obtained by using a combination of cine gradient-echo or steady-state free precession and cine phase-contrast sequences. In addition to providing measurements of peak velocity and flow, MR imaging is the standard of reference for evaluation of ventricular function, which can be a critical factor in determining when surgical intervention is indicated. Improvements in cardiac MR imaging technology have been particularly striking in the past few years, and these developments can easily be applied to the examination of cardiac valves. The authors briefly describe the pathophysiology of valvular disease, discuss standard MR techniques for qualitative and quantitative evaluation of valvular lesions, and illustrate these concepts with several case studies. PMID- 12886891 TI - The lesson of history is how little we appreciate and/or remember. PMID- 12886892 TI - SARS (severe acute respiratory syndrome). PMID- 12886893 TI - HIV vaccine trial results are an important step forward in developing an effective vaccine, say WHO and UNAIDS. PMID- 12886894 TI - Cancer patients suffering pain unduly. PMID- 12886895 TI - Impact of the introduction of a combined Haemophilus B conjugate vaccine and hepatitis B recombinant vaccine on vaccine coverage rats in a large West Coast health maintenance organization. AB - We evaluated the effect of introduction of a combined hepatitis B-Haemophilus influenzae b vaccine on vaccine coverage rates in a large health maintenance organization and found a modest improvement in up-to-date immunization status. PMID- 12886896 TI - Immunologic memory with no detectable bactericidal antibody response to a first dose of meningococcal serogroup C conjugate vaccine at four years. AB - Fourteen children with no detectable bactericidal antibody response to a first dose of meningococcal C conjugate vaccine at 4 years of age were given a booster dose of the same vaccine 2 years later. A rapid 1000-fold rise in postimmunization bactericidal antibody titers, a measured either 7 or 14 days later, suggested previous immunologic priming. PMID- 12886897 TI - Increased incidence of pneumococcal bloodstream infections. AB - Pneumococcal bloodstream infections in pediatric patients rose from 3.6 to 14.8 episodes per 100,000 from 1986 to the year 2000 (P < 0.001) in a population of approximately 650,000 inhabitants (16% <16 years of age). Incidence in the group <24 months of age increased from 7.4 to 78.6 episodes per 100,000 per year (P < 0.001). Overall 75% of the serogroups are included in the new heptavalent vaccine. PMID- 12886898 TI - Hemophagocytic syndrome after Kawasaki disease. AB - Hemophagocytic syndrome (HPS) is a rare and life-threatening disease in which a generalized histiocytic proliferation results in hemophagocytosis and up regulation of inflammatory cytokines. This syndrome has been associated with infections, malignancy, drugs and immunologic triggers such as Kawasaki disease (KD). We describe the clinical and laboratory features of two children with HPS after KD and review the three previously reported pediatric cases of recrudescence of HD leading to HPS. PMID- 12886899 TI - Vibrio vulnificus necrotizing fasciitis of the calf presenting with compartment syndrome. AB - We describe a 17-year-old boy with congenital spherocytosis and iron overload who presented with compartment syndrome of the calf as the initial manifestation of Vibrio vulnificus infection after minor trauma in a contaminated fish pond. The disease was complicated by necrotizing fasciitis requiring above the knee amputation. Childhood diseases associated with iron overload pose an increased risk for complicated V. vulnificus infections. PMID- 12886900 TI - Fatal lactic acidosis and mimicking Guillain-Barre syndrome in an adolescent with human immunodeficiency virus infection. AB - We report a case of antiretroviral therapy-related fatal lactic acidosis occurring in a vertically infected HIV-positive 17-year-old patient. While receiving antiretroviral therapy with stavudine, didanosine, tenofovir and amprenavir, the patient developed severe acidosis and rapid neuromuscular and respiratory failure mimicking Guillain-Barre syndrome. PMID- 12886901 TI - Refinement and reduction in production of genetically modified mice. PMID- 12886902 TI - Research funding organisations and consumer involvement. PMID- 12886903 TI - National responses to HIV/AIDS: the importance of understanding context. PMID- 12886904 TI - [Statistical analysis on papers and authors for Chinese Journal of Hepatology]. PMID- 12886905 TI - Abstracts of the 25th Annual Carrell-Krusen Symposium. Dallas, Texas, USA. February 27-March 1, 2003. PMID- 12886906 TI - [Control of infectious diseases: achievements and tasks]. PMID- 12886907 TI - [Control of diabetes melittus in China: a pressing situation]. PMID- 12886908 TI - The bottom line. PMID- 12886909 TI - A systematic chart review of the nature of psychiatric adverse events in children and adolescents treated with selective serotonin reuptake inhibitors. AB - OBJECTIVE: Despite a rapidly growing literature on the efficacy of the selective serotonin reuptake inhibitors (SSRI) in the treatment of juvenile psychiatric disorders, relatively little is described about emotional, behavioral, and cognitive adverse effects associated with their use. To this end we completed a retrospective analysis of medical charts to determine the incidence, nature, and clinical correlates of treatment emergent adverse effects in the behavioral, cognitive, and emotional domains. METHODS: We systematically evaluated the medical charts of children treated with SSRI for depressive or obsessive compulsive disorders for a mean (+/- SD) of 26.9 + 20.8 months to determine the incidence, nature, and clinical correlates of treatment emergent psychiatric adverse events (PAE). Charts were reviewed for diagnoses, type and dose of SSRI and adjunct medication, specific type of PAE, and time to onset and offset of PAE. RESULTS: In total, 82 charts of children and adolescents (mean age 12.2 +/- 3.2 years) were examined. PAE occurred in 22% of children and were most commonly related to disturbances in mood. PAE were not associated with psychiatric diagnosis(es), age, sex, concurrent medications, doses or specific serotonin reuptake inhibitors. The onset of PAE was observed typically 3 months after SSRI exposure (median = 91 days). Although PAE diminished with SSRI discontinuation, those that emerged early in treatment diminished significantly more rapidly than those that emerged later (median offset was 10 and 49 days, respectively). Re exposure to an SSRI resulted in another PAE in 44% (n = 18) of the group. CONCLUSION: Based on the retrospective review of medical charts, youth receiving SSRI appear to be at risk for treatment emergent PAE and recurrence with re exposure to an SSRI. Prospective longer term studies evaluating the course and prognosis of youths manifesting PAE to SSRI are necessary. PMID- 12886910 TI - [Proceedings and abstracts of the 9th National Congress of the Italian Society of Adolescent Medicine. Catanzaro, Italy, October 24-26, 2002]. PMID- 12886911 TI - Proceedings of the European Symposium on Congenital Toxoplasmosis. 10-11 October 2002, Paris, France. PMID- 12886912 TI - Factors affecting the spread of bupivacaine in the adult thoracic paravertebral space. AB - Factors affecting the spread of bupivacaine in the paravertebral space were investigated in patients undergoing paravertebral nerve blocks for the treatment of chronically painful conditions. Injections of bupivacaine 0.5%, 10-15 ml mixed with depomedrone up to 80 mg were repeated at 2-wk intervals up to a maximum of four times. A blinded observer mapped out the subsequent distribution of sensory loss to cold on both sides of the torso at 5-min intervals after each injection. Age, sex, height and weight did not correlate with the block; nor did injectate volume, mass of bupivacaine, previous posterolateral thoractomy and spread of radiocontrast. Injections repeated at 2-wk intervals in the same patient resulted in different degrees of spread that were unrelated to one another. Time to peak onset of blockade was 40 min in 95% of patients. A single bolus of bupivacaine produces a safe but unpredictable block. Yet to be defined physical properties and anatomical factors are probably key determinants of the spread of bupivacaine in the paravertebral space. This single bolus technique may be better supplanted by a reversion to the older multiple level injection technique. PMID- 12886913 TI - The information requested by patients prior to giving consent to anaesthesia. AB - Patients in a teaching hospital in Scotland were given the opportunity to ask for further information relating to their peri-operative anaesthetic management. One investigator visited all of the patients pre-operatively and asked a standard list of anaesthetic-related questions. Our objectives were to determine what additional information patients would request before giving their consent to anaesthesia. The majority of patients (n=469, 67%) had no further questions at a point when so-called consent could have been obtained. Of the questions asked, 209 (66%) were related to anaesthesia and 93 (30%) to the proposed surgery. Only two patients in the group studied requested a full explanation of their peri operative anaesthetic management. The question most commonly asked related to the duration of surgery, with less priority being given to questions relating to complications of anaesthesia. PMID- 12886914 TI - [Proceedings and abstracts of the IV International Symposium on Renal Osteodystrophy. Oviedo, 6-7 June 2002]. PMID- 12886915 TI - A comparison of intra-operative or postoperative exposure to music--a controlled trial of the effects on postoperative pain. AB - The effect of intra-operative compared to postoperative music on postoperative pain was evaluated in a controlled trial. In all, 151 patients undergoing day case surgery for inguinal hernia repair or varicose vein surgery under general anaesthesia were randomly allocated to three groups: group 1 listened to music intra-operatively, group 2 listened to music postoperatively and group 3, the control group, listened to 'white noise'. The anaesthetic and postoperative analgesic techniques were standardised. Pain was assessed using a numeric rating scale (0-10) and patients requirements for postoperative morphine, paracetamol and ibuprofen was recorded. The effect of music on nausea, fatigue and anxiety was also investigated. The results showed that patients exposed to music intra operatively or postoperatively reported significantly lower pain intensity at 1 and 2 h postoperatively and patients in the postoperative music group required less morphine at 1 h compared to the control group. No differences were noted in the other variables. This study demonstrates that there is a short-term pain reducing effect of music therapy however, the beneficial effects do not differ if the patient is exposed to music intra-operatively or postoperatively. PMID- 12886916 TI - [Proceedings of the centenary celebration of the foundation of the "Clinica del Lavoro Luigi Devoto" of Milan. 4 March 2002]. PMID- 12886917 TI - A comparison of cyclizine, ondansetron and placebo as prophylaxis against postoperative nausea and vomiting in children. AB - Nausea and vomiting is a relevant and common problem with unfavourable sequelae in children undergoing some plastic surgery procedures. There is a lack of anti emetic trials performed in children, with only a few investigating the roles of the older anti-emetic agents such as cyclizine compared with newer ones such as ondansetron. This randomised, controlled, double-blind study examined the effectiveness of a single dose of ondansetron (0.1 mg x kg-1), cyclizine (20 mg) and placebo (normal saline) in the prevention of postoperative nausea and vomiting in 150 children (mean age 3.6 years) undergoing plastic genitourinary procedures. Rates of previous postoperative nausea and vomiting and motion sickness were comparable across the groups. Postoperative vomiting was significantly reduced with ondansetron prophylaxis (p = 0.006) but there was no detectable anti-emetic effect with cyclizine. Furthermore, cyclizine caused pain on injection (p < 0.001). PMID- 12886918 TI - Variability in determination of point of needle insertion in peripheral nerve blocks: a comparison of experienced and inexperienced anaesthetists. AB - Accurate identification of surface landmarks is essential for the successful performance of peripheral nerve blocks. The variability between experienced and inexperienced practitioners in identifying anatomical landmarks has not been studied previously. Anaesthetists were asked to identify the point of needle insertion for posterior lumbar plexus and sciatic nerve blocks on a volunteer using a standard textbook description. The chosen point for needle insertion was described in terms of X and Y co-ordinates, measured in millimetres, from a zero reference point marked on a volunteer's back. Fifteen experienced and 22 inexperienced anaesthetists took part in the study. The lumbar plexus block mean [range] values for the X, Y co-ordinates were 80 [62-108], 66 [46-86] and 92 [49 150], 62 [0-131] in the experienced and inexperienced groups, respectively. The sciatic nerve block X, Y co-ordinates were 77 [62-99], 70 [49-89] and 68 [29 116], 62 [26-93] in the experienced and inexperienced groups, respectively. The variance for the point of needle insertion was significantly greater in the inexperienced group (p <0.01) for both the lumbar plexus and sciatic nerve blocks. We conclude that with increasing experience, there is decreased variability in determining the point of needle insertion using anatomical landmarks. PMID- 12886919 TI - Day of surgery cancellations after nurse-led pre-assessment in an elective surgical centre: the first 2 years. AB - We describe a nurse-led pre-assessment system at an elective surgical centre. A targeted referral system was used by trained nurses to direct referrals to a supervising consultant anaesthetist or to the surgical team. Of 2726 patients pre assessed in the first 2 years, 105 patients (3.9%) were cancelled or postponed for medical optimisation after pre-assessment. There were 137 cancellations (5.0%) on the day of surgery, despite pre-assessment, but only 36 were for anaesthetic or medical reasons. Only eight of these 36 were considered a 'failure' of the pre-assessment system. These results are much better than the cancellation rate of about 11% in the Trust as a whole. There were 18 transfers of patients postoperatively from the elective centre to another hospital. A review suggested that four of these transfers could have been reasonably predictable from the patients' medical history. We conclude that a pre-assessment clinic has an important role to play in minimizing cancellations on the day of surgery and also in reducing the number of patients transferred to other hospitals. This last conclusion has an important implication for the planning of systems in hospitals that perform only elective surgery. PMID- 12886920 TI - Personal assistance services on the job. PMID- 12886921 TI - Epilepsy programs across the states. PMID- 12886922 TI - [Challenges to the development of China's occupational medicine]. PMID- 12886923 TI - [Epidemiological study of renal syndrome-hemorrhagic fever: progress, problems and prospects]. PMID- 12886924 TI - [Basic study of child lead poisoning in China: problems]. PMID- 12886925 TI - Finance issue brief: continuity of care: year end report-2002. AB - When a health care provider leaves or is terminated by a managed care plan, how long can patients continue to receive covered treatment from that provider for a life-threatening condition, pregnancy, disability or other circumstance that requires continuous medical care? Lawmakers in 36 states have answered that question. PMID- 12886926 TI - Finance issue brief: direct access: year end report-2002. AB - Consumer demand for a less restrictive referral system has urged legislators, as well as the managed care industry and those in the medical field, to seek the best solution to the problem. The point of service (POS) plan was an early attempt by managed care plans to provide more freedom for enrollees. But POS plans are not always available or cost effective. Several of the larger managed care organizations, in an effort to improve enrollee and provider satisfaction, have eliminated or streamlined their referral processes. The American Medical Association has recommended that primary care and other medical specialty organizations collaborate in developing guidelines for specialty referrals, urging health plans to allow direct access in network without financial penalty as long as the access conforms with the established guidelines. In the meantime, legislators have responded to constituents by enacting measures that mandate direct access for specific types of care or standing referrals for ongoing treatment. PMID- 12886927 TI - [How to treat for Dubreuilh's melanoma?]. PMID- 12886928 TI - Finance issue brief: freedom of choice: year end report-2002. AB - Should managed care plans have the ability to limit an individual's "freedom of choice" when it comes to selecting a provider? As part of the anti-managed care movement of the early 1990s, 22 states enacted freedom of choice laws; the majority apply only to pharmacies. PMID- 12886929 TI - [Hygienic monitoring system in China]. PMID- 12886930 TI - [Protection of workers' health and the development of labour hygiene]. PMID- 12886931 TI - Finance issue brief: health care claims payment: prompt payment: year end report 2002. AB - Since the mid 1990's state legislators and regulators have worked to resolve the complex issue of timely payment of health care claims. They have been challenged with bridging the communication gap between provider and payor and forced to address such base problems as what determines a correctly billed service. As time has progressed it is ever apparent that the completion of payment for services is dependent on many variables, not just simply timely processing of a claim. PMID- 12886932 TI - Finance issue brief: HMO medical directors: year end report-2002. AB - Medical directors of managed care organizations have the ultimate responsibility of deciding the treatments for which a health plan will pay. Cognizant of consumer concerns over the power inherent in the position, states are seeking to inject more accountability into the decision-making process. Lawmakers have begun to grapple with whether medical directors should be required to hold a medical license from the state in which they work and, by extension, whether they should be under the jurisdiction of the state medical board. PMID- 12886933 TI - Finance issue brief: insurer liability: year end report-2002. AB - When a health plan denies payment for a procedure on grounds that it is not medically necessary or when it refuses a physician-ordered referral to a specialist, has it crossed the line from making an insurance judgment to practicing medicine? If the patient suffers harm as a result of the decision, is the plan liable for medical malpractice? Those were questions 35 states considered in 1999, and at least 32 states are grappling with this year as they seek to respond to physician and patient pressure to curb the power of the managed care industry. Traditionally, health insurers have been protected by state laws banning "the corporate practice of medicine," which means the patient's only recourse is to sue under a "vicarious liability" theory. Now, however, lawmakers are debating legislation to extend the scope of malpractice liability beyond individual practitioners to insurance carriers and plans themselves. PMID- 12886934 TI - Finance issue brief: long-term care insurance: year end report-2002. AB - A 1996 federal law made it more attractive for states to consider long-term care insurance, and states have responded by implementing policies to make the purchase of these long-term care coverage more affordable and consumer-friendly. At present, policy makers continue to debate the future role of private long-term care insurance in subsidizing the increasing demand for long-term care services. PMID- 12886935 TI - Finance issue brief: managed care report cards: year end report-2002. PMID- 12886936 TI - [Molecular epidemiology in the study of diseases]. PMID- 12886937 TI - Prophages and bacterial genomics: what have we learned so far? AB - Bacterial genome nucleotide sequences are being completed at a rapid and increasing rate. Integrated virus genomes (prophages) are common in such genomes. Fifty-one of the 82 such genomes published to date carry prophages, and these contain 230 recognizable putative prophages. Prophages can constitute as much as 10-20% of a bacterium's genome and are major contributors to differences between individuals within species. Many of these prophages appear to be defective and are in a state of mutational decay. Prophages, including defective ones, can contribute important biological properties to their bacterial hosts. Therefore, if we are to comprehend bacterial genomes fully, it is essential that we are able to recognize accurately and understand their prophages from nucleotide sequence analysis. Analysis of the evolution of prophages can shed light on the evolution of both bacteriophages and their hosts. Comparison of the Rac prophages in the sequenced genomes of three Escherichia coli strains and the Pnm prophages in two Neisseria meningitidis strains suggests that some prophages can lie in residence for very long times, perhaps millions of years, and that recombination events have occurred between related prophages that reside at different locations in a bacterium's genome. In addition, many genes in defective prophages remain functional, so a significant portion of the temperate bacteriophage gene pool resides in prophages. PMID- 12886938 TI - Molecular histochemical analysis of estrogen receptor alpha and beta expressions in the mouse ovary: in situ hybridization and Southwestern histochemistry. AB - It is well known that estrogen plays important roles in the female reproductive organs, including the ovary, through estrogen receptors (ERalpha and ERbeta). It is suggested that selective effects of estrogen on a discrete set of genes may reside in promoters that are differentially responsive to ERalpha and ERbeta, while the relative biological significance of ERalpha and ERbeta is unclear. Therefore, for a better understanding of the physiological and pathological states of ovarian cells, it seems to be essential to analyze the cellular expression of ERalpha and ERbeta, at both the protein and mRNA levels. For this purpose, we have optimized the protocols for immunohistochemistry and nonradioactive in situ hybridization, respectively, using the mouse ovary. These techniques have been established as useful histochemical methods for the localization of specific protein and/or mRNA expression at the cellular level. Moreover, we also attempted to evaluate ERs as binding proteins to estrogen responsive elements, using Southwestern histochemistry, which permits us to analyze the localization in situ of transcription regulatory proteins. By employing these methods, more precise understanding of specific gene expression and evaluation of its biological significance in the ovary would be possible. PMID- 12886939 TI - Cell biology of human ovarian surface epithelial cells and ovarian carcinogenesis. AB - Most epithelial ovarian carcinomas have been suggested to arise from the ovarian surface epithelium, which covers an ovary as a layer of flat to cuboidal cells. The epithelium is physiologically involved in follicular rupture and the subsequent repair of the follicle wall during reproductive age. Invagination and inclusion cysts are formed in the cortical stroma after cyclic ovulation. Consequently, ovulation may cause a loss of integrity of the surface epithelium followed by stepwise sequence of genetic alteration. Inclusion cysts are actually more common in ovaries contralateral to those containing malignant epithelial tumors than in control ovaries. Human ovarian surface epithelial cells exhibit a gland formation in coculture with endometrial stromal cells in an estrogen-rich environment. The phenotypic plasticity of these cells shares a mesenchymal property when they are cultured on two layers of extracellular matrix and collagen gel. As an in vitro study of ovarian carcinogensis, several neoplastic cell lines were recently established from the surface epithelial cells of the human ovary. SV 40 large T-antigen transfection into the epithelial cells induced some immortalized cell lines, one of which showed anchorage-independent growth and tumor formation in athymic mice. The tumors were histologically undifferentiated carcinoma. These cell lines may lead to insights into the preneoplastic and early stages of epithelial ovarian carcinomas. To understand the pathogenesis of epithelial ovarian cancer, specifically designed studies of ovarian surface epithelium and the related structural changes encountered after ovulation and these existing in ovarian carcinomas are required. PMID- 12886940 TI - Effect of epicardial or biventricular pacing to prolong QT interval and increase transmural dispersion of repolarization. PMID- 12886942 TI - Current awareness on yeast. PMID- 12886941 TI - Low triiodothyronine and cardiovascular disease. PMID- 12886944 TI - Behavioral phenotyping enhanced--beyond (environmental) standardization. AB - It is basic biology that the phenotype of an animal is the product of a complex and dynamic interplay between nature (genotype) and nurture (environment). It is far less clear, however, how this might translate into experimental design and the interpretation of animal experiments. Animal experiments are a compromise between modelling real world phenomena with maximal validity (complexity) and designing practicable research projects (abstraction). Textbooks on laboratory animal science generally favour abstraction over complexity. Depending on the area of research, however, abstraction can seriously compromise information gain, with respect to the real world phenomena an experiment is designed to model. Behavioral phenotyping of mouse mutants often deals with particularly complex manifestations of life, such as learning, memory or anxiety, that are strongly modulated by environmental factors. A growing body of evidence indicates that current approaches to behavioral phenotyping might often produce results that are idiosyncratic to the study in which they were obtained, because the interactive nature of genotype-environment relationships underlying behavioral phenotypes was not taken into account. This paper argues that systematic variation of genetic and environmental backgrounds, instead of excessive standardization, is needed to control the robustness of the results and to detect biologically relevant interactions between the mutation and the genetic and environmental background of the animals. PMID- 12886945 TI - The fallacy of behavioral phenotyping without standardisation. AB - Behavioral phenotyping of mutant mice is a new and challenging task for the behavioral neuroscientist. Therefore, standardisation of the experimental conditions is required to permit comparisons between the results of experiments within and between laboratories. Once mutation-induced behavioral changes have been identified, phenotyping of mouse mutants should be performed along a systematic trajectory, which allows for an in-depth characterisation of the mutant under investigation. PMID- 12886946 TI - Harnessing the mouse to unravel the genetics of human disease. AB - Complex traits, i.e. those with multiple genetic and environmental determinants, represent the greatest challenge for genetic analysis, largely due to the difficulty of isolating the effects of any one gene amid the noise of other genetic and environmental influences. Methods exist for detecting and mapping the Quantitative Trait Loci (QTLs) that influence complex traits. However, once mapped, gene identification commonly involves reduction of focus to single candidate genes or isolated chromosomal regions. To reach the next level in unraveling the genetics of human disease will require moving beyond the focus on one gene at a time, to explorations of pleiotropism, epistasis and environment dependency of genetic effects. Genetic interactions and unique environmental features must be as carefully scrutinized as are single gene effects. No one genetic approach is likely to possess all the necessary features for comprehensive analysis of a complex disease. Rather, the entire arsenal of behavioral genomic and other approaches will be needed, such as random mutagenesis, QTL analyses, transgenic and knockout models, viral mediated gene transfer, pharmacological analyses, gene expression assays, antisense approaches and importantly, revitalization of classical genetic methods. In our view, classical breeding designs are currently underutilized, and will shorten the distance to the target of understanding the complex genetic and environmental interactions associated with disease. We assert that unique combinations of classical approaches with current behavioral and molecular genomic approaches will more rapidly advance the field. PMID- 12886947 TI - The ultimate chip shot: can microarray technology deliver for neuroscience? AB - The use of cDNA and oligonucleotide microarrays, or 'chips', is emerging as a powerful, new technology in the field of neuroscience for examining gene expression in a high-throughput fashion. The application of microarray technology to the study of brain and behavior has lagged behind other areas of biology such as cancer and yeast genetics due to the challenges presented by the heterogeneous and complex organization of the nervous system. This review provides a brief overview of available microarray technology as well as a description of experimental considerations in planning and implementing a neuroscience-based array study. Successful implementation of microarray technology within the field of neuroscience will provide a molecular approach to studying systems neurobiology, leading to insights into areas ranging from fundamental questions of developmental neurobiology to neurological and psychiatric disorders. PMID- 12886948 TI - Expression profiling identifies strain-specific changes associated with ethanol withdrawal in mice. AB - Mice that exhibit characteristics of physical dependence following ethanol exposure serve as useful models of alcoholism in humans. The DBA/2J and C57BL/6J inbred strains differ in their behavioral response to ethanol withdrawal. Alterations in gene expression are believed to underlie neuroadaptation to ethanol dependence and tolerance. Therefore, the differences in ethanol withdrawal severity observed between the DBA/2J and C57BL/6J strains may be related to differential regulation of gene expression. We have used cDNA microarrays to determine the gene expression profile in the hippocampus of DBA/2J and C57BL/6J mice during withdrawal after chronic and acute ethanol exposure. Of the 7634 genes surveyed, approximately 2% were consistently differentially expressed by at least 1.4-fold in DBA/2J mice during chronic ethanol withdrawal. Less than 1% of the genes showed altered expression in C57BL/6J mice under the same conditions, or in DBA/2J mice during acute ethanol withdrawal. Strain- and treatment-specific patterns of altered expression were observed for multiple genes associated with the Janus kinase/signal transducers and activators of transcription and the mitogen activated protein kinase pathways. Genes associated with both pathways are regulated in DBA/2J mice during chronic ethanol withdrawal, and to a lesser extent during acute ethanol withdrawal. Only those genes associated with the mitogen-activated protein kinase (MAPK) pathway exhibited changes in expression in C57BL/6J mice during ethanol withdrawal. Furthermore, genes associated with retinoic acid-mediated signaling show differential expression exclusively in C57BL/6J mice. These findings represent significant differences in cellular adaptation to ethanol between the DBA/2J and C57BL/6J strains. PMID- 12886949 TI - Preweaning enrichment has no lasting effects on adult hippocampal neurogenesis in four-month-old mice. AB - Since both living in an enriched environment and physical activity stimulate hippocampal neurogenesis in adult mice, we endeavored to examine whether pre weaning enrichment, a sensory enrichment paradigm with very limited physical activity, had similar effects on neurogenesis later in life. Mice were removed from the dams for periods of increasing length from post-natal day 7 to 21, and exposed to a variety of sensory stimuli. At the age of 4 months, significant differences could be found between previously enriched and nonenriched animals when spontaneous activity was monitored. Enriched mice moved longer distances, and spent more time in a defined center zone of the open field. Adult neurogenesis was examined by labeling proliferating cells in the dentate gyrus with bromodeoxyuridine (BrdU). Cell proliferation, survival of the newborn cells, and net neurogenesis were similar in both groups. Volumetric measurements and stereological assessment of total granule cell counts revealed no difference in size of the dentate gyrus between both groups. Thus, in contrast to postweaning enrichment, preweaning enrichment had no lasting measurable effect on adult neurogenesis. One of the parameters responsible for this effect might be the lack of physical activity in preweaning enrichment. As physical activity is an integral part of postweaning enrichment, it might be a necessary factor to elicit a neurogenic response to environmental stimuli. The result could also imply that baseline adult hippocampal neurogenesis is independent of the changes induced by preweaning enrichment and might not contribute to the sustained types of plasticity seen in enriched animals. PMID- 12886950 TI - Behavioral profiles of inbred strains on novel olfactory, spatial and emotional tests for reference memory in mice. AB - Studying the behavior of genetic background strains provides important information for the design and interpretation of cognitive phenotypes in mutant mice. Our experiments examined the performance of three commonly used strains (C57BL/6J, 129S6, DBA/2J) on three behavioral tests for learning and memory that measure very different forms of memory, and for which there is a lack of data on strain differences. In the social transmission of food preference test (STFP) all three strains demonstrated intact memory for an odor-cued food that had been sampled on the breath of a cagemate 24 hours previously. While C57BL/6J and 129S6 mice showed good trace fear conditioning, DBA/2J mice showed a profound deficit on trace fear conditioning. In the Barnes maze test for spatial memory, the 129S6 strain showed poor probe trial performance, relative to C57BL/6J mice. Comparison of strains for open field exploratory activity and anxiety-like behavior suggests that poor Barnes maze performance reflects low exploratory behavior, rather than a true spatial memory deficit, in 129S6 mice. This interpretation is supported by good Morris water maze performance in 129S6 mice. These data support the use of a C57BL/6J background for studying memory deficits in mutant mice using any of these tasks, and the use of a 129S6 background in all but the Barnes maze. A DBA/2J background may be particularly useful for investigating the genetic basis of emotional memory using fear conditioning. PMID- 12886951 TI - Methods for comparing sources of strand compositional asymmetry in microbial chromosomes. AB - Significant compositional biases in bacterial chromosomes have been explained by replication- and transcription-coupled repair mechanisms, the latter causing GC skew to indicate the direction of replication when gene polarity is correspondingly entrained. Correlations between indicators of replication direction, skew, and transcription polarity are computed for the complete nucleotide sequences of 20 microbial chromosomes and interpreted through statistical tests. A second quantitative method, previously applied to the first complete draft of the Escherichia coli K12 genome, characterizes the sequences by average skew and net skew due to replication. These methods generally agree in finding the coexistence of replication- and translation-coupled effects and in identifying atypical sequences in which one influence is clearly dominant. The replication-dominated class is exemplified by two chlamydial sequences and the transcription-dominated class by three archaea. The preference for leading-strand transcription in two mycoplasmas is stronger than the skew implies. These concordant methods provide an objective framework for comparing sources of strand compositional asymmetry and interpreting skew diagrams. PMID- 12886952 TI - Genome-wide expression analysis of the responses to nitrogen deprivation in the heterocyst-forming cyanobacterium Anabaena sp. strain PCC 7120. AB - A heterocyst is a terminally differentiated cell of cyanobacteria which is specialized in dinitrogen fixation. Heterocyst differentiation in Anabaena sp. strain PCC 7120 is triggered by deprivation of combined nitrogen in the medium. Although various genes that are upregulated during heterocyst differentiation have been reported, most studies to date were limited to individual or a small number of genes. We prepared microarrays in collaboration with other members of the Anabaena Genome Project. Here we report on the genome-wide expression analysis of the responses to nitrogen deprivation in Anabaena. Many unidentified genes, as well as previously known genes, were found to be upregulated by nitrogen deprivation at various time points. Three main profiles of gene expression were found: genes expressed transiently at an early stage (1-3 hr) of nitrogen deprivation, genes expressed transiently at a later stage (8 hr), and genes expressed when heterocysts are formed (24 hr). We also noted that many of the upregulated genes were physically clustered to form 'expressed islands' on the chromosome. Namely, large, continuous genomic regions containing many genes were upregulated in a coordinated manner. This suggests a mechanism of global regulation of gene expression that involves chromosomal structure, which is reminiscent of eukaryotic chromatin remodelling. The possible implications of this global regulation are discussed. PMID- 12886953 TI - Structural analysis of a Lotus japonicus genome. IV. Sequence features and mapping of seventy-three TAC clones which cover the 7.5 mb regions of the genome. AB - Using the sequence information of expressed sequences tags (ESTs), cDNAs and genes from Lotus japonicus and other legumes, 73 TAC (transformation-competent artificial chromosomes) clones were selected from a genomic library of L. japonicus accession MG-20, and their nucleotide sequences were determined. The length of the DNA sequenced in this study was 7,455,959 bp, and the total length of the DNA regions sequenced so far is 26,167,443 bp together with the nucleotide sequences of 183 TAC clones previously reported. By similarity searches against the sequences in protein and EST databases and prediction by computer programs, a total of 699 potential protein-encoding genes with known or predicted functions, 163 gene segments and 267 pseudogenes were assigned to the newly sequenced regions. Based oil the nucleotide sequences of the clones, simple sequence repeat length polymorphism (SSLP) or derived cleaved amplified polymorphic sequence (dCAPS) markers were generated, and each clone was located onto the linkage map of two accessions of L. japonicus, Gifu B-129 and Miyakojima MG-20. The sequence data, gene information and mapping information are available through the World Wide Web at http://www.kazusa.or.jp/lotus/. PMID- 12886954 TI - Identification of a human cDNA sequence which encodes a novel membrane-associated protein containing a zinc metalloprotease motif. AB - We report the cloning and characterization of a human cDNA predicted to encode a novel hydrophobic protein containing four transmembrane domains and a zinc metalloprotease motif, HEXXH, between the third and fourth transmembrane domains, and have named the molecule metalloprotease-related protein-1 (MPRP-1). The MPRP 1 gene was localized to chromosome 1-p32.3 by radiation hybrid mapping, and Northern blot analysis revealed expression in many organs, with strong expression in the heart, skeletal muscle, kidney and liver. Immunohistochemical analyisis showed that MPRP-1 was localized in the endoplasmic reticulum (ER), and not in the Golgi compartment. Fragments of DNA encoding a segment homologous to the HEXXH motif of MPRP-1 are widely found in bacteria, yeast, plants, and animals. These results suggest that the MPRP-1 may have highly conserved functions, such as in intracellular proteolytic processing in the ER. PMID- 12886955 TI - High-throughput production of recombinant antigens for mouse KIAA proteins in Escherichia coli: computational allocation of possible antigenic regions, and construction of expression plasmids of glutathione-S-transferase-fused antigens by an in vitro recombination-assisted method. AB - Since the end of 2001, we have conducted a project to isolate and determine entire sequences of mouse cDNA clones which encode the polypeptides corresponding to human KIAA proteins. Towards the ultimate goal of this project to clarify the biological functions of KIAA genes, we have set production of antibodies against mouse KIAA gene products based on their sequence information as the next important stage. As the first step, we developed a high-throughput system utilizing shotgun clones generated during entire sequencing of mouse KIAA cDNAs. The system consists of the following three parts: (1) Shotgun clones encoding regions suitable for production of antigens were selected using a newly developed browser system; (2) the protein-coding sequences of the selected shotgun clones were transferred into an expression vector by in vitro recombination-assisted method in a 96-well format, and expressed as glutathione S-transferase fusion proteins in Escherichia coli; and (3) the solubility of the recombinant antigens were preliminarily assessed in a small-scale culture and then large-scale production and purification was performed using glutathione-affinity beads or retrieval from polyacrylamide gels depending on their solubility. Using these systems, we successfully produced and purified 400 antigens for production of mKIAA antibodies to date. PMID- 12886956 TI - The lessons of SARS. PMID- 12886957 TI - Vibrio vulnificus infection: clinical manifestations, pathogenesis, and antimicrobial therapy. AB - There has been a dramatic increase in the number of reported cases of infection due to Vibrio vulnificus in Taiwan. Although the organism has been etiologically implicated in a variety of clinical syndromes, most cases of V. vulnificus infection are categorized as primary bacteremia, skin and soft tissue infection. The mortality was up to 50% in septic patients, most of them dying within 48 h of admission. In most of the cases involving V. vulnificus infection have underlying disease, particularly liver cirrhosis. The pathogenesis may attribute to several virulent factors, such as lipopolysaccharide, capsular lipopolysaccharide, cytolysin, metalloprotease and siderophore. Tetracycline was suggested as the drug of choice based on an animal study. Our previous in vitro data showed that cefotaxime and minocycline acted synergistically in inhibiting V. vulnificus. Furthermore, another in vivo animal study indicated that therapy using combined with cefotaxime and minocycline was distinctly more advantageous than therapy with the single antibiotic regimen for the treatment of severe experimental murine V. vulnificus infection. Recently, we also demonstrated that the newer fluoroquinolones, as single agents were as effective as the combination therapy both in vitro and in vivo. PMID- 12886958 TI - Concurrent infections by two dengue virus serotypes among dengue patients in Taiwan. AB - The co-circulation of multiple dengue virus serotypes in the same region has been reported in several countries in Southeast Asia as well as in Central and South America for decades. Although outbreaks involving more than one serotype of dengue virus have been reported in Taiwan since 1987, concurrent infection in the same individual by multiple serotypes of dengue virus have never been identified. Using a modified multiplex reverse transcription-polymerase chain reaction assay, we detected and determined the serotypes of 21 dengue patients during an outbreak in southern Taiwan in 2000. While either dengue type 2 or type 3 virus was found in most cases, 2 of the 21 cases were concurrent infections by both dengue type 2 and dengue type 3 viruses. This was further confirmed by sequence analysis of the amplified products. This study reports for the first time that concurrent infections occur in Taiwan, and suggests that future virologic surveillance of dengue outbreak in Taiwan should take this into consideration. PMID- 12886959 TI - Vaccination trials against Taenia solium eggs in pigs injected with frozen oncospheres of T. solium or Taenia saginata asiatica. AB - In this study, 12 Small-Ear-Miniature pigs aged 142 to 185 days were used to determine whether pigs injected with nonviable oncospheres of Taenia solium or Taenia saginata asiatica can become resistant to the challenge of viable eggs of T. solium. The 12 pigs were equally divided into 4 groups: 3 experimental groups in which each pig was injected subcutaneously with a mixture of 0.2 mL complete Freund's adjuvant and 10(4)/0.2 mL nonviable Taiwan/Asian Taenia, Indonesia Taenia, or T. solium oncospheres, and 1 control group in which each pig was injected subcutaneously with 0.2 mL phosphate buffer solution and 0.2 mL complete Freund's adjuvant. Each pig was orally inoculated with 10000 viable T. solium eggs 1 month later. The infection rates were 100% (2/2), 100% (3/3), 33% (1/3), and 100% (3/3) and cysticerci recovery rates were 1.3% (254/20000), 1.2% (371/30000), 0.01% (4/30000), and 8.6% (2,577/30000), respectively. Except for the location of 72 cysticerci located in the viscera, 3134 cysticerci were recovered from the muscles. In the experimental groups, 4 cysticerci recovered were viable and the remaining 625 were either calcified or degenerated. However, 2567 cysticerci recovered from the control group remained viable and only 10 were calcified or degenerated. The results indicate that in addition to the vaccine of T. solium, those of Taiwan Taenia and Indonesia Taenia can also induce high crossing immunologic reactions against T. solium infection. PMID- 12886960 TI - Relationship between human lymphocyte antigen-B27 and clinical features of psoriatic arthritis. AB - Psoriatic arthritis is a chronic destructive joint disease. About 40% of psoriatic arthritis patients are positive for human lymphocyte antigens (HLA) B27. This study investigated the relationship between HLA-B27 and clinical manifestations and prognosis in psoriatic arthritis patients. Demographic, clinical, and laboratory data were analyzed from 41 psoriatic arthritis patients with regular follow-ups. The mean percentage of HLA-B27 in psoriatic arthritis was about 39%. Positive HLA-B27 was associated with an increased risk of development of sacroillitis (relative risk 8.75; p<0.01) but not peripheral arthritis (p=0.925). Psoriatic arthritis patients with psoriatic nail disease (41.5% vs 2.4%, p<0.01) and distal interphalangeal joints involvement (26.8% vs 3.4%, p<0.05) had significantly increased risk of developing deformed joints. Psoriatic arthritis patients with positive HLA-B27 tend to develop deformed joints (p=0.068) as well as having elevated levels of C-reactive protein (p=0.072), although these results did not attain significance. HLA-B27 antigen may serve as a useful predictive marker for the development of sacroiliitis in Taiwan. PMID- 12886961 TI - Antibiotic resistance of pathogenic bacteria from odontogenic infections in Taiwan. AB - The purpose of this study was to evaluate the susceptibility of facultative and anaerobic odontogenic infectious flora to various antibiotics. We assessed 178 bacterial strains isolated from 74 patients with odontogenic infections. The E test was used to determine susceptibility. The microbial flora was predominantly facultative gram-positive organisms and anaerobic gram-negative bacilli. The results of antimicrobial susceptibility test showed that ampicillin resistance was found with a very high level of minimum inhibitory concentration (MIC) in approximately one third of Fusobacterium nucleatum, Prevotella intermedia, Peptostreptococcus micros, and Eikenella corrodens isolates (MIC > or = 8 microg/mL). Greater activity was generally noted with amoxicillin than with ampicillin, but even beta-lactamase inhibitor incorporated amoxicillin showed resistance in more than 10% of all groups except viridans group streptococci and Porphyromonas gingivalis. Tetracycline and erythromycin were considerably less active against the majority of the tested bacterial strains, while minocycline and doxycycline exerted strong antimicrobial activity and could inhibit strains grown at a very low concentration. Among all the tested antibiotics, travofloxacin appears to be a promising drug expressing the highest activities (MIC90 < or = 1 microg/mL), and was regarded as a potent bactericidal drug in odontogenic infections. PMID- 12886962 TI - Clinical characteristics of children with influenza A virus infection requiring hospitalization. AB - From September 1997 to March 2002, a total of 84 children were admitted to Chang Gung Children's Hospital due to influenza A virus infection. Influenza A virus infection was documented in 61 cases by viral isolation from throat and in 23 cases by serologic studies. The mean age of patients was 43.8 months, ranging from 20 days to 16 years. Forty-one (49%) patients were male. Lower respiratory tract infection (53 of 84 cases) was the most common clinical manifestation, occurring predominantly in children younger than 5 years (49 of 53 cases). The types of lower respiratory tract infection included bronchiolitis/bronchopneumonia in 33 cases, pneumonia in 17, and croup in 3. Central nervous system dysfunction was noted in 26 patients, predominantly in older children (18 of 26 cases). This included encephalopathy in 11 cases, encephalitis in 10, aseptic meningitis in 2, psychosis in 1, febrile convulsions in 1, and acute disseminated encephalomyelitis in 1. Gastrointestinal symptoms were mild in most patients. Diarrhea occurred in 18.4% of the children younger than 5 years, compared with only 8.4% of the older children. By contrast, abdominal pain was more common in older children (16.7%) than in younger children (6.7%). Ten children had leukocytosis (white blood cell > or = 15000 /microL) and 9 of them were younger than 5 years. Eleven children had a C-reactive protein level greater than 100 mg/L and 10 of them were younger than 5 years. The mean duration of fever and hospitalization were 4.6 +/- 2.8 days and 7.4 +/- 5.7 days, respectively. The clinical outcomes were excellent in all but 1 patient who died from intractable pulmonary hemorrhage. The frequency and duration of hospitalization due to influenza A virus is much greater than generally thought in Taiwan, suggesting an urgent need for educational programs to increase awareness of the characteristics and risks for this illness. PMID- 12886963 TI - Independent prognostic factors for fatality in patients with invasive vibrio cholerae non-O1 infections. AB - To identify independent prognostic factors for fatality, 73 patients with a total of 75 episodes of invasive Vibrio. cholerae non-O1 infections treated from July 1998 through October 2001 at 2 medical centers were retrospectively studied. The demographic, laboratory, and clinical information of these patients were collected and analyzed. The overall mortality rate was 36%. Multivariate analysis revealed that severe liver cirrhosis (p=0.003; odds ratio [OR], 14.12, with 95% confidence interval [CI] 2.49-79.91), malignancy (p=0.034; OR, 3.9, with 95% CI 1.11-13. 7), and steroid use (p=0.011; OR, 12.37, with 95% CI 1.79-85.49) were independent risk factors for fatality. These findings suggest that patients at high risk of fatality should be hospitalized and aggressively treated when V. cholerae non-O1 infections develop, and that public education on how to avoid exposure to V. cholerae non-O1 is important for the high-risk population. PMID- 12886964 TI - Impact of antibiotic use on fungus colonization in patients hospitalized due to fever. AB - Invasive candidiasis is an opportunistic infection that occurs in medical and surgical patients and carries a high mortality rate. Because its process always follows from colonization to amplification and to invasion, understand the colonization status is important to understand the likelihood of invasion. Screening for fungal colonization was performed with specimens from urine, throat, anus, and skin in 314 patients admitted for fever both before and after antibiotics treatment was administered. Throat (45%) and anus (43%) had the highest colonization rates. Only 7% of patient had fungal colonization on the skin. Candida albicans was the most frequently colonized species in throat (79%) and anus (70%). Colonized fungal species on skin were diverse, including Candida parapsilosis (33%), C. albicans (29%), and molds (24%). Sex (M:F ratio, 49.7:50.3) was not a factor in fungal colonization, but aging was associated with increased colonization rate. Forty-five patients received antibiotics treatment for more than 2 weeks and second surveillance cultures were taken at the end of the second week. Antibiotics treatment was associated with increased frequency of colonization (p=0.02), but the fungal species distribution pattern remained unchanged after antibiotic treatment. PMID- 12886965 TI - Brain abscess: clinical analysis of 53 cases. AB - Fifty-three cases of brain abscess were treated at Mackay Memorial Hospital from January 1991 through December 2001. The ages ranged from 2 weeks to 84 years, with a peak at 40 to 50 years (11/53, 21%). The male to female ratio was 1.8:1 (34 males, 19 females). The most common presenting symptoms were fever (30/53, 57%), headache (29/53, 55%), and changed mental status (24/53, 45%). The duration of symptoms before hospitalization ranged from several hours to 2 months. A shorter duration of symptoms was associated with poor outcome. The common predisposing factors were otic infection (10/53, 19%), penetrating head trauma and neurosurgery (10/53, 19%), and bacterial endocarditis (5/53, 9%). The leading underlying diseases were diabetes mellitus (12/53, 23%) and/or liver cirrhosis (6/53, 11%), and both were independently associated with increased risk of mortality. Computed tomographic scanning and magnetic resonance imaging facilitated early diagnosis and proper management. Surgical intervention was used together with antibiotics in 33 (62%) of 53 patients in whom the average abscesses diameter was 3.75 cm (range, 2-6 cm). The remaining 20 (38%) patients whose average abscesses diameter was 2.3 cm (range, 1-3.5 cm) were treated with antibiotics only. Culture of material drained from abscesses isolated 27 microorganisms from 19 (58%) of the 33 patients, 81% (22/27) of which were aerobic and 19% (5/27) anaerobic bacteria. The most common pathogen was alpha hemolytic Streptococcus spp. (6/27, 22%). Most of the patients with Klebsiella pneumoniae isolated from brain abscess, cerebrospinal fluid, and blood cultures were diabetic. A high mortality rate (9/20, 45%) was found in patients with medical treatment. A high index of suspicion is needed for the early diagnosis of brain abscess, particularly in patients with predisposing factors. In this series, early diagnosis using computed tomography and/or magnetic resonance scanning, optimal timing of surgery, and appropriate use of antibiotics were associated with improved outcome. PMID- 12886966 TI - Methotrexate pneumonitis in a patient with rheumatoid arthritis. AB - Methotrexate pneumonitis is an unpredictable and life-threatening side effect of methotrexate therapy. Early diagnosis, cessation of methotrexate, and treatment with corticosteroids and/or cyclophosphamide are important in the management of patients with methotrexate pneumonitis. Methotrexate pneumonitis has not been reported in patients of Chinese ethnicity. We report a case of methotrexate pneumonitis in a Taiwan patient with rheumatoid arthritis who presented with acute nonproductive cough, dyspnea, fever, severe hypoxemia, and rapid progression to respiratory failure. Chest roentgenogram demonstrated bilateral diffuse interstitial and alveolar infiltration. Thoracoscopic biopsy with wedge resection of the upper lobe of the right lung was performed and the histologic findings of the biopsy specimen were consistent with bronchiolitis obliterans with organizing pneumonia. Rapid improvement of methotrexate pneumonitis was achieved after pulse therapies of methylprednisolone and cyclophosphamide and daily use of prednisolone. PMID- 12886967 TI - Chromobacterium violaceum bacteremia: a case report. AB - Chromobacterium violaceum is confined in tropical and subtropical regions, which can cause life-threatening disease. It is the only Chromobacterium species that is pathogenic to humans. Because of its rarity, clinicians often do not appreciate its importance when it is isolated. We report a fulminate fatal case of C. violaceum bacteremia in a 20-year-old male Taiwanese. The clinical manifestations were fever and abdominal pain, followed by shock and pulmonary septic embolism. Emergent laparotomy identified acute appendicitis with rupture. Flomoxef sodium was administered immediately. However, his condition deteriorated rapidly and he died within 48 h after the onset of illness. Two sets of blood culture yielded C. violaceum. Physicians should be aware of the occurrence of this infection in summer season. PMID- 12886968 TI - Aspergillus flavus epidural abscess and osteomyelitis in a diabetic patient. AB - A 63-year-old man had a history of diabetes mellitus for more than 10 years and took oral hypoglycemic agents regularly. He visited Taipei Veterans General Hospital with the complaint of progressive weakness in all 4 limbs and neck pain for 6 months. Computed tomography of the cervical spine revealed increased soft tissue density in the epidural space from C2 to C5 with cord compression. Surgical decompression was done and Aspergillus flavus was isolated from the inflammatory tissue. He was initially treated with oral itraconazole 200 mg 3 times per day for 4 days and then twice daily. Later, the treatment regimen was shifted to intravenous amphotericin B 25 mg/d. He died of intraventricular hemorrhage and complicated fungal meningoencephalitis 2 weeks postlaminectomy. This case reminds us that a prolonged history of back pain accompanied with peripheral neuropathy in diabetic patients should raise the suspicion of Aspergillus epidural abscess. Prompt aggressive diagnostic testing and management is needed to improve the likelihood of a good outcome of these patients. PMID- 12886969 TI - Culture-to-culture physical interactions causes the alteration in red and infrared light stimulation of Escherichia coli growth rate. AB - Escherichia coli MC1061 cells were irradiated at 660 and 900 nm, incubated in M9 and LB media with the use of a specially constructed device, and assayed for growth rate. There was a reduction of growth rate stimulation when the irradiated culture was cultivated jointly with the non-irradiated one. In the same time, the irradiated culture extended the invigorate effect on the non-irradiated one. It is proposed that the effects observed were mediated by culture-to-culture physical interactions. PMID- 12886970 TI - Intravenous immunoglobulin as adjunctive therapy for juvenile spasms. AB - Intravenous immunoglobulin has been reported to be an effective treatment for infantile spasms. Juvenile spasms are electrically and clinically similar to infantile spasms but occur in a later age group. We retrospectively reviewed the charts of five children (aged 4.5-11.5 years) at our institution. Their primary seizure type was juvenile spasms and they were treated with a single inpatient course of intravenous immunoglobulin (400 mg/kg/day intravenously for 5 consecutive days) on an adjunctive basis. Seizure frequency was determined from parental reports. By 3 months after treatment, improvement (a 50-92% reduction in seizure frequency) was noted in four patients; sustained benefit was seen in three patients for up to 12 months. One patient showed no response at 3 and 6 months and had an increase in seizure frequency at 1 year. We conclude that single-course intravenous immunoglobulin can be effective as adjunctive therapy for juvenile spasms and that benefit can persist for many months. However, larger controlled, prospective clinical trials are needed to validate this unconventional treatment modality for specific seizure types such as juvenile spasms. PMID- 12886971 TI - Evoked potentials in spinal muscular atrophy. AB - Visual evoked potentials, brainstem evoked responses, and somatosensory evoked potentials were evaluated in 22 children with spinal muscular atrophy, types I and II. Eleven of the children had the severe form of spinal muscular atrophy (type I) and 11 children had the intermediate form (type II). The results of visual evoked potentials, brainstem evoked responses, and somatosensory evoked potentials were compared with those obtained in a control group. Statistical analysis showed abnormalities in the different sensory modalities. A significant increase in the visual evoked potential latencies was observed and was found more often in patients with spinal muscular atrophy type I. Alterations of the somatosensory thalamocortical responses were also observed, as well as a delay in the central conduction time. Although spinal muscular atrophy is usually considered to be a purely motor disorder involving neurons of the spinal anterior horn and nuclei of the lower cranial nerves, lesions of the posterior roots, spinal ganglia, ascending tracts, lateral geniculated corpus, and thalamus have been reported. Our results suggest that sensory neuron degeneration occurs more commonly in spinal muscular atrophy than previously thought and that this process probably develops more slowly than motoneuron degeneration. Such degeneration may be associated with brain atrophy. PMID- 12886972 TI - Dissociation of motor maturation. AB - We prospectively acquired clinical data regarding the presentation, evaluation, and developmental progress of all patients identified with dissociated motor maturation to define their clinical outcomes. Children (N = 8) referred for evaluation of suspected cerebral palsy because of delayed sitting or walking and identified to have dissociated motor maturation were followed with serial clinical examination. All displayed the characteristic "sitting on air" posture while held in vertical suspension and had otherwise normal developmental assessments. This posture is composed of the hips held in flexion and abduction with the knees extended and feet plantar or dorsiflexed. Three children were initially evaluated at 10 months of age owing to absence of sitting and five other children were evaluated at a mean of 14 months (range 12-19 months) owing to inability to stand. Follow-up evaluations were conducted over a mean of 10.5 months (range 5-34 months). Five children were born prematurely at 34 to 36 weeks gestation. Denver Developmental Screening Test and general and neurologic examinations were normal except to note hypotonia in six children and the "sitting on air" posture in all of the children. Four children have older siblings or parents who "walked late" (after 15 months). On average, the children attained sitting by 8 months (range 7-10 months). One child did not crawl prior to independent walking, two children scooted rather than crawled, and five children crawled at an average of 13.5 months (range 10-16 months). All children cruised by a mean of 18 months (range 16-21.5 months) and attained independent walking by 20.1 months (range 18-25 months). Neuroimaging and serum creatine kinase enzyme testing were normal in two children who were tested. These eight children conform to the syndrome of dissociated motor maturation. The "sitting on air" posture serves as a diagnostic sign and anticipated excellent prognosis, but follow-up is required to ensure a normal outcome. PMID- 12886973 TI - Utility of electroencephalography in the evaluation of common neurologic conditions in children. AB - The objective of this study was to assess the utility of electroencephalography (EEG) in the evaluation of common neurologic conditions in children. The EEG recordings of 534 consecutive children (aged < 20 years) were prospectively read by a certified pediatric neurologist. Common diagnostic indications included the following: clinical seizures (33.8%), definite epilepsy (31.2%), attention deficit hyperactivity disorder (ADHD) (9.1%), headache (8%), syncope (3.5%), learning disabilities (2%), tic disorders (1.4%), and sleep disorders (1.1%). Overall, 63.8% of EEG records were normal, slowing background activity was noted in 6.1%, ADHD was noted in 35.3% (definite epilepsy), and epileptiform activity was noted in 37.1% of the cases of definite epilepsy and 13.2% of the clinically suspected cases. Epileptiform activity was rarely found in the patients without epilepsy. All EEG records of children with syncope (n = 19) and headache (n = 43) were normal. These findings indicate that although EEG plays an important diagnostic role in epilepsy, interictal EEG is being overused during evaluation of various neurologic disorders in children, and a normal EEG is highly predictable in children with nonepileptic conditions. PMID- 12886974 TI - Atonic seizures in survivors of childhood cancer. AB - Atonic seizures are part of some childhood epilepsy syndromes and can also result from acquired central nervous system insult. Of 93 survivors of childhood cancer with uncontrolled seizures, 10 (11%) with atonic seizures form the basis for this report. Seven survived acute leukemia, and three survived brain tumors. The median age at cancer diagnosis was 9 months (range 2-52). Nine received systemic and six intrathecal chemotherapy; cranial irradiation was given to seven. All have learning difficulties, abnormal brain magnetic resonance images, and an abnormal electroencephalogram; six are developmentally delayed. All have difficult-to-control seizures, but six improved on myoclonus-specific drugs accelerated development in two younger patients. Atonic seizures in cancer survivors are associated with difficult-to-control seizures and pervasive cognitive impairments. Young age at cancer diagnosis and cranial irradiation may predispose the patient to development of this syndrome. Recognition and appropriate treatment of atonic seizures in cancer survivors could improve seizure control, neurologic development, and quality of life. PMID- 12886975 TI - Acute transverse myelitis in children: clinical course and prognostic factors. AB - The objective of this study was to describe the clinical course of acute transverse myelitis in children, to identify prognostic factors, and to compare our findings with published data Twenty-four children, aged 2 to 14 years and admitted with a diagnosis of acute transverse myelitis, were studied. Clinical features and results of investigations were collected at admission and during the course of the disease. Motor, sphincter, and global outcomes were compared with those in the main adult and pediatric series. During the initial phase, the most common presenting symptoms were pain (88%) and fever (58%). Motor loss preceded sphincter dysfunction in two thirds of patients and became bilateral in half of the patients. When maximal deficit was achieved (plateau), the patients presented a combination of sensory, motor, and sphincter dysfunctions without radicular involvement The motor loss consistently involved the lower limbs but was inconsistent and moderate in the upper limbs. The mean duration of the plateau was 1 week. The recovery phase was characterized by a progressive improvement of all deficits. Sphincter dysfunction improved more slowly than did the other deficits. A full recovery was achieved by 31% of the patients; minimal sequelae were present in 25% and mild to severe sequelae in 44%. An unfavorable outcome was associated with complete paraplegia (P = .03) and/or a time to maximal deficit shorter than 24 hours (P = .005). A favorable outcome was associated with a plateau shorter than 8 days (P = .03), the presence of supraspinal symptoms (P = .01), and a time to independent walking shorter than 1 month (P = .01). The course of acute transverse myelitis in children proceeds through three stages, an initial phase, a plateau, and a recovery phase, each characterized by specific clinical features. The global outcome was favorable in 56% of patients. Several prognostic factors were identified. PMID- 12886976 TI - Relative influence of epileptic seizures and of epilepsy syndrome on cognitive function. AB - Cognitive impairment is frequently observed in children with epilepsy. We aimed at addressing to what extent cognitive function is affected by paroxysmal epileptiform activity with or without clinical seizures or by clinical features characteristic of the epilepsy syndrome. To this purpose, combined electroencephalographic (EEG) recording and cognitive testing (IQ and reaction times) were performed in 28 children. Frequent epileptiform EEG discharges significantly reduced reaction time, as did the occurrence of seizures during cognitive testing. Syndrome-related factors tended to affect cognitive functions as well: children with generalized epilepsy and high average seizure frequency obtained lower scores. Linear regression analysis showed that stable aspects of cognitive function, as reflected in intelligence level, are most closely related to the severity of the epilepsy syndrome (average seizure frequency), whereas transient aspects of cognitive function, such as reaction times, are related to the occurrence of epileptiform EEG discharges. This suggests that seizures have a direct effect on transient cognitive aspects, which can accumulate and result in effects on intelligence level. PMID- 12886977 TI - Lies, damned lies, and medical experts: the abrogation of responsibility by specialty organizations and a call for action. AB - The festering problem of medical expert witnesses proffering false testimony in court remains. A review of the many proposed reforms to resolve this chronic and outrageous practice has indicated little progress despite a resolution by the American Medical Association enunciating that medical expert witness testimony is effectively the practice of medicine. A questionnaire was sent to 36 specialty organizations to determine whether they had established guidelines, position statements, policies, or bylaws that govern the disciplinary management of their members who testify falsely. Responses were obtained from all 36 specialty organizations and showed that over 80% had no definitive disciplinary policies to deal with this egregious practice. This finding is all the more surprising given repeated judicial and other exhortations to professional organizations to "police themselves." To address the abrogation of responsibility by specialty organizations, new stringent disciplinary steps are proposed to exorcise the despicable practice of false expert witness testimony from the practice of medicine. PMID- 12886978 TI - Intra-arterial tissue plasminogen activator for thrombosis complicating cerebral angiography in a 17-year-old girl. AB - Few reports describe the use of intraarterial recombinant tissue plasminogen activator to treat intracranial thrombosis in children. A 17-year-old girl with a history of prior venous thrombosis developed a left middle cerebral artery thrombus during diagnostic cerebral angiogram. Therapy with intra-arterial tissue plasminogen activator was initiated. An immediate follow-up angiogram demonstrated recanalization, and diffusion-weighted magnetic resonance imaging 9 hours later showed no evidence of infarction. Following the angiogram, femoral artery thrombosis developed. Treatment with supratherapeutic levels of heparin, localized delivery of intra-arterial tissue plasminogen activator, embolectomy, danaparoid, and dipyridamole failed to re-establish perfusion to the lower leg, and below the knee amputation was required. Neurologic examination remained normal 1 year later. Cerebral damage was avoided with the use of emergency intra arterial tissue plasminogen activator for cerebral artery thrombosis in this child. PMID- 12886979 TI - Benign "setting sun" phenomenon in full-term infants. AB - I report two normally developed infants showing benign" setting sun" phenomenon. A 2(2-12)-year-old boy and a 7-year-old boy, who were born without any complications at full term, developed brief episodes of downward gazing during sucking and crying after birth However, there were no other clinical or laboratory findings, and they developed normally. The phenomenon was not visible until 6 months and 7 months, respectively. The "setting sun" phenomenon usually indicates underlying severe brain damage and can also be seen, although rarely, in healthy full-term infants until 1 to 5 months. However, the benign "setting sun" phenomenon might exist until 6 or 7 months of age in normal infants. PMID- 12886980 TI - Infantile Sandhoff's disease: multivoxel magnetic resonance spectrosecopy findings. AB - Sandhoff's disease is a rare, genetic lysosomal storage disease leading to delayed myelination or demyelination. Although neuroimaging findings in this disease have been reported previously, magnetic resonance spectroscopy findings have not been reported. In this report, we present magnetic resonance imaging and magnetic resonance spectrscopy features of two cases with Sandhoff's disease. Magnetic resonance spectroscopy revealed findings indicating widespread demyelination in both cases and neuroaxonal loss and anaerobic metabolism in the second case. Magnetic resonance spectroscopy could provide useful information in the explanation of the clinical picture of cases with Sandhoff's disease. PMID- 12886981 TI - Unusual early-onset Huntingtons disease. AB - Huntington's disease is an autosomal dominant progressive neurodegenerative disorder characterized by involuntary movements, cognitive decline, and behavioral disorders leading to functional disability. In contrast to patients with adult onset, in which chorea is the major motor abnormality, children often present with spasticity, rigidity, and significant intellectual decline associated with a more rapidly progressive course. An unusual early-onset Huntington's disease case of an 11-year-old boy with severe hypokinetic/rigid syndrome appearing at the age of 2.5 years is presented. Clinical diagnosis was confirmed by polymerase chain reaction study of the expanded IT-15 allele with a compatible size of 102 cytosine-adenosine-guanosine repeats L-Dopa mildly ameliorated rigidity, bradykinesia, and dystonia. We conclude that Huntington's disease should be included in the differential diagnoses of regressive syndromes of early childhood. PMID- 12886982 TI - Photosensitivity during the hypersomnic phase in a patient with Kleine-Levin syndrome. AB - The case of an adolescent with Kleine-Levin syndrome is presented who exhibited a consistent and predictable photoparoxysmal self-limited response to intermittent photic stimulation during all relapses of his hypersomnic phase. A possible cause of this association that has not been previously reported is speculated based on observations concerning the two disorders. PMID- 12886983 TI - Multifocal myoclonus following group A streptococcal infection. AB - Movement disorders as postinfectious manifestation of group A streptococcal infections have been reported and are thought to occur on an autoimmune basis. We describe an unusual case of multifocal myoclonus following strep throat infection. Clinical description and chart review were the method used. A 10-year old boy developed focal myoclonus involving his right arm and shoulder 1 week after streptococcal throat infection treated with penicillin. His magnetic resonance image was normal, and he initially responded to clonazepam but did not sustain a response. The myoclonus spread to involve all limbs and the trunk becoming multifocal over the next few weeks. He did not have choreoathetosis of Syndenham's chorea. He was given one course of intravenous immunoglobulin and became asymptomatic after treatment. He remained symptom free for 8 months following intravenous immunoglobulin treatment. Various symptoms have been reported following group A streptococcal infections in children. These neurobehavioral abnormalities may be mediated through antineuronal antibodies. Our case demonstrates multifocal myoclonus as a poststreptococcal autoimmune phenomenon. To our knowledge, only two other cases of poststreptococcal myoclonus have been reported in the literature. Recognition of this unusual condition as a manifestation of autoimmune poststreptococcal disease in children is essential to avoid overinvestigation and to ensure early treatment. PMID- 12886984 TI - Stroke owing to noncompaction of myocardium. AB - Noncompaction of myocardium is a rare and recently defined entity that may cause cardioembolism during childhood. We report an 18-month-old girl with noncompaction of the left ventricular myocardium presenting with fatal cardioembolic stroke. The patient had a high factor VIII level, which is known to cause an increased tendency to thromboembolic events. To our knowledge, this is the youngest case with stroke associated with noncompaction of the myocardium. Patients with noncompaction should be considered for prophylactic antithrombotic treatment to prevent mortality and morbidity owing to systemic thromboembolic events, especially if they carry additional risk factors that make them prone to hypercoagulation. PMID- 12886985 TI - Idiopathic intracranial hypertension in the pediatric population. PMID- 12886986 TI - Prostaglandin E2-mediated regulation of immunoglobulin G2 via interferon gamma. AB - BACKGROUND: Patients with localized aggressive periodontitis (LAgP) produce elevated levels of IgG2 both in vivo and in vitro. Responses to the periodontitis associated pathogens Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis are dominated by IgG2, and these IgG2 responses are associated with reduced extent and severity of disease. Little is known about regulation of the IgG2 subclass, although prostaglandin E2 (PGE2) (a mediator known to polarize responses toward Th-2) and interferon (IFN)-gamma (a Th-1 mediator) both promote IgG2 production. This unusual relationship prompted the hypothesis that, in certain circumstances, PGE2 enhances rather than inhibits IFN-gamma production. METHODS: To test this hypothesis, indomethacin (IND)-treated peripheral blood mononuclear cell (PBL) cultures from healthy volunteers were stimulated with pokeweed mitogen (PWM), and the cultures were manipulated by adding PGE2, rIFN gamma, rIL-Ialpha, rIL-1beta, rIL-6, or rIL-12. Production of IgG1, IgG2, IFN gamma, and PGE2 was monitored by enzyme-linked immunosorbent assay. RESULTS: Indomethacin treatment inhibited IgG1 and IgG2 production, and PGE2 restored both immunoglobulins in PWM-stimulated cultures. Remarkably, addition of IFN-gamma also restored IND-suppressed IgG2 but not IgG1. In contrast, addition of rIL (interleukin)-1alpha, rIL-1beta, rIL-6, or rIL-12 did not restore IgG2 responses. Furthermore, IND suppressed IFN-gamma production and PGE2 increased IFN-gamma levels. Kinetic studies indicate that PGE2 production occurred on the first day of culture, followed by IFN-gamma two days later. CONCLUSIONS: These findings support the concept that in certain circumstances, PGE2 production can lead to increased IFN-gamma and that this IFN-gamma selectively promotes IgG2 responses. These data suggest that the elevated PGE2 observed in LAgP patients may contribute to increased IFN-gamma production and help explain elevated IgG2 responses. PMID- 12886987 TI - Novel isolation of alkaline phosphatase-positive subpopulation from periodontal ligament fibroblasts. AB - BACKGROUND: Periodontal ligament fibroblasts (PDLFs) are the cells essential for periodontal regeneration. PDLFs comprise a heterogeneous cell population and consist of several cell subsets that differ in their function. It is known that PDLFs produce osteoblast-related extracellular matrix proteins and show higher alkaline phosphatase (ALP) activity compared with gingival fibroblasts (GFs), implying that PDLFs have osteogenic characterisitics. The aim of the present study was to isolate the osteogenic population of PDLFs according to their expression of ALP. METHODS: PDLFs and gingival fibroblasts were separated into two populations, ALP-positive and ALP-negative, with an immunomagnetic method using a monoclonal antibody against human bone type ALP and magnetic beads conjugated with a secondary antibody. Expression of basic fibroblast growth factor (bFGF) receptor and transforming growth factor (TGF)-beta receptor was investigated in these two populations. Osteoblast-related molecules, osteocalcin, and bone sialoprotein; ALP activity; and effect of bFGF on proliferation were also compared. RESULTS: Effective separation was confirmed in both PDLFs and GFs by flow cytometry. The expression of FGF receptor (FGFR) and TGF-beta receptor was significantly higher in ALP-positive PDLFs than in ALP-negative PDLFs. ALP positive PDLFs also expressed higher mRNA levels of osteocalcin and bone sialoprotein compared with ALP-negative PDLFs. The mitogenic effect of bFGF on ALP-positive PDLFs was greater than that of ALP-negative PDLFs. CONCLUSIONS: These results indicate that osteoblastic and/or cementoblastic PDLF subsets could be isolated from the PDLF populations using an immunomagnetic method. Magnetic isolation of PDLFs may be a useful tool to obtain the cells which will potentially induce mineralization on the root surface. PMID- 12886988 TI - Effect of recombinant human bone morphogenetic protein-4 with carriers in rat calvarial defects. AB - BACKGROUND: Bone morphogenetic proteins (BMPs) are being evaluated as candidates for periodontal and bone regenerative therapy. However, the research on recombinant human bone morphogenetic protein-4 (rhBMP-4) has been insufficient to evaluate its capacity to enhance bone formation and its carrier system. The purpose of this study was to evaluate the bone regenerative effect of rhBMP-4 delivered with an absorbable collagen sponge (ACS) or beta-tricalcium phosphate (beta-TCP). We also compared the potential of beta-TCP to that of ACS as a carrier system for rhBMP-4. METHODS: Eight-mm calvarial critical-sized defects were created in 100 male Sprague-Dawley rats. The animals were divided into 5 groups of 20 animals each. The defects were treated with rhBMP-4/ACS (rhBMP-4 at 0.05 mg/ml), rhBMP-4/beta-TCP (rhBMP-4 at 0.05 mg/ml), ACS alone, beta-TCP alone, or left untreated for surgical control. The rats were sacrificed at 2 or 8 weeks postsurgery, and the results were evaluated radiodensitometrically, histologically, and histomorphometrically. RESULTS: The results of radiodensitometric analysis were as follows: the rhBMP-4/ACS and the rhBMP-4/beta TCP groups were more radiopaque than other groups at both 2 and 8 weeks (P < 0.01). The histologic observations were as follows: in the rhBMP-4/ACS and the rhBMP-4/beta-TCP groups, new bone was evident at the defect sites at 2 weeks and 8 weeks. The results of histomorphometric analysis were as follows: the rhBMP 4/ACS and the rhBMP-4/beta-TCP groups had more bone (%) than other groups at both 2 and 8 weeks (P < 0.01). CONCLUSIONS: Surgical implantation of rhBMP-4/ACS may be used to support bone regeneration in the rat calvarial critical-sized defect, and rhBMP-4/beta-TCP may be able to regenerate bone in the rat calvarial critical sized defect without complication. In addition, both ACS and beta-TCP may be considered as available carriers for rhBMP-4. PMID- 12886989 TI - Detection of Tannerella forsythensis (Bacteroides forsythus) and porphyromonas gingivalis by polymerase chain reaction in subjects with different periodontal status. AB - BACKGROUND: The aim of this study was to determine the prevalence of Tannerella forsythensis (formerly Bacteroides forsythus) and Porphyromonas gingivalis in subgingival plaque samples by using polymerase chain reaction (PCR), and to assess the relationship of these bacteria with different categories of periodontal disease and health. METHODS: Subjects were distributed into 3 groups according to their periodontal diagnosis: group 1, periodontally healthy (N = 10); group 2, periodontitis with probing depth < or = 5 mm (N = 10); group 3, periodontitis with probing depth > 5 mm (N = 10). The subjects in groups 2 and 3 had healthy and diseased periodontal sites. Subgingival plaque samples were obtained using paper points inserted into periodontal pockets (diseased sites) and into healthy gingival sulci (healthy sites) of the same subject. RESULTS: The distribution of bacteria differed in healthy and diseased sites. T. forsythensis (B. forsythus) was not detected in any sample from healthy sites in any group but was detected in 70% and 100% of diseased sites in groups 2 and 3, respectively. P. gingivalis was detected in only one sample from a healthy site (group 2), and in the diseased sites, its prevalence was 40% (group 2) and 90% (group 3). In addition, T. forsythensis (B. forsythus) and P. gingivalis were both detected in 30% and 90% of the diseased sites in groups 2 and 3, respectively. CONCLUSION: These results indicate a possible association between periodontal disease and the presence of T. forsythensis (B. forsythus) and/or P. gingivalis. PMID- 12886990 TI - Subgingival microflora in Turkish patients with periodontitis. AB - BACKGROUND: No information exists on periodontitis-associated subgingival microbiota from Turkey. We determined the occurrence, interspecies relationships, and clonal characteristics for a group of periodontal bacteria in a Turkish study population. METHODS: Subgingival microbial samples were obtained from patients with localized (LAgP, N = 18) or generalized (GAgP, N = 17) types of aggressive periodontitis, generalized chronic periodontitis (GCP, N = 14), and non periodontitis subjects (N = 20). Culture methods were used to recover 6 periodontal bacterial species and yeasts, and a polymerase chain reaction technique was used to detect Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis. Intraspecies characterization of A. actinomycetemcomitans was carried out by serotyping and genotyping. RESULTS: All species, except for Micromonas micros (formerly Peptostreptococcus micros) occurred more frequently (P < 0.05) in periodontitis than non-periodontitis subjects. Detection frequencies for Tannerella forsythensis (formerly Bacteroides forsythus) and Campylobacter rectus differed among the periodontitis subgroups; the lowest frequency occurred in LAgP. The mean proportions of A. actinomycetemcomitans, P. gingivalis, and C. rectus were higher (P < 0.008) in GAgP than in non-periodontitis subjects. Significant positive associations were seen between 7 of the 22 possible combinations (P < 0.05). A. actinomycetemcomitans serotype c (34%) and non-serotypeable isolates (34%) were the most common antigenic types among the 305 strains analyzed. Eleven arbitrarily primed (AP)-PCR genotypes were distinguished among 273 isolates from 29 subjects. Yeasts were found in 23% of the 69 subjects. CONCLUSIONS: The results on the Turkish study population were generally in line with earlier reports on the occurrence and interspecies relationships of certain bacteria in periodontitis. However, A. actinomycetemcomitans was not overrepresented in LAgP, and the serotype distribution resembled that reported from the East. The high frequency of non-serotypeable isolates suggests local characteristics of the species. PMID- 12886991 TI - Crown lengthening in mandibular molars: a 5-year retrospective radiographic analysis. AB - BACKGROUND: Crown lengthening procedures are a significant part of the periodontist's armamentarium. In order to recreate the clinical space lost to caries or trauma necessary for prosthetic restoration, osseous surgery is often required. If the procedure is not carefully planned, it may result in furcation involvement of multirooted teeth. METHODS: Twenty-six subjects with 26 mandibular molar teeth requiring crown lengthening procedures prior to prosthetic crown placement were evaluated. Nineteen subjects with 24 prosthetic crowns on lower molars which had not undergone crown lengthening were included as control teeth. Bite-wing radiographs prior to surgery (for the test group) or placement of the crown (control group) and 5 years after completion of the prosthesis were compared and analyzed. RESULTS: Of the 26 test teeth, 10 teeth (38.5%) were found to have radiographic evidence of furcation involvement, whereas none of the control teeth developed furcation invasions. CONCLUSION: A critical distance from the furcation (CDF) of 4 mm was established as a landmark under which, if surgery was performed on mandibular molars, chances of furcation involvement in the future were very high. PMID- 12886992 TI - Quantitative evaluation of the vasculature and fibronectin localization in gingival connective tissue of smokers and non-smokers. AB - BACKGROUND: It has been shown that tobacco is a significant risk factor for periodontal disease. The reason for decreased gingival bleeding in smokers is not known. The aim of the present study was to demonstrate the localization of fibronectin in the extracellular matrix of the inflamed gingiva of smokers and non-smokers and to evaluate the chronic effect of smoking on the gingival microvessel number (NVES) and vascular surface density (VSD). METHODS: Seventy four (74) adult patients with periodontitis were included in this investigation. Of these patients, 38 were smokers and 36 were non-smokers. Probing depths (PD), papillary bleeding index (PBI) of both groups, and the smoking habits of the smokers were recorded. Biopsy specimens obtained during periodontal surgery were prepared to evaluate fibronectin distribution and to quantitate the vasculature. RESULTS: The mean VSD values of smokers and non-smokers were 6.721 +/- 1.845 and 5.721 +/- 3.118 (mean +/- SD), and the mean NVES of smokers and non-smokers was 31.582 +/- 11.810 and 30.145 +/- 15.442, respectively. The difference between the mean PD and PBI values of the 2 groups was not statistically significant. The location of the biopsy specimen, whether in the anterior or posterior area of the mouth, did not lead to any statistically significant differences between the groups. In addition, the number of years smoked and the tobacco brand used did not result in statistically significant differences. The most intense staining of fibronectin was observed beneath the epithelium around the blood vessels of the uppermost region of the connective tissue, showing a fibrillar and diffuse distribution; however, there was no significant difference between smokers and non-smokers. CONCLUSION: The results of the present study indicate that smoking does not affect the vascular surface density, number of vessels per mm2 stroma, or fibronectin distribution in subepithelial gingival connective tissue. PMID- 12886993 TI - Mandibular bone remodeling under a choline-deficient diet: a histomorphometric study in rats. AB - BACKGROUND: A deficiency of lipotropic factors in the rat induces renal, hepatic, and/or hematic damage. The aim of the present study was to evaluate the effect of a choline-deficient diet and refeeding on mandibular bone remodeling. METHODS: Fifty Wistar rats were divided into 5 groups: group 1 (G1): control diet for 15 days; group 2 (G2): choline-deficient diet for 15 days; group 3 (G3): control diet for 30 days; group 4 (G4): choline-deficient diet for 30 days; and group 5 (G5): choline-deficient diet for 15 days and control diet for 15 days. All animals were sacrificed by ether overdose. The mandibles were resected, radiographed, decalcified, processed, and embedded in paraffin. Bucco-lingually oriented sections were obtained at the level of the interradicular bone of the medial roots of the left first molar, and stained with hematoxylin and eosin (H & E). Bone tissue density and bone remodeling were determined histomorphometrically. Body weight, food intake, hematocrit, and hemoglobinemia were also recorded. RESULTS: Microscopic observation revealed that osteogenesis was lower in rats fed a choline-deficient diet, at both 15 and 30 days, and that this decrease did not revert with a control diet. Histomorphometric evaluation showed 37% and 27% reduction in bone tissue density at 15 and 30 days, respectively, and a 30% decrease in bone formation at 30 days, compared to controls. CONCLUSION: In this experimental model, a choline-deficient diet led to altered bone remodeling as observed by a marked reduction in osteogenesis. PMID- 12886994 TI - Effect of ER:YAG and diode laser irradiation on the root surface: morphological and thermal analysis. AB - BACKGROUND: The aim of the present study was to compare the effects of Er:YAG and diode laser treatments of the root surface on intrapulpal temperature after scaling and root planing with hand instruments. METHODS: Fifteen extracted single rooted teeth were scaled and root planed with hand instruments. The teeth were divided into 3 groups of 5 each and irradiated on their buccal and lingual surfaces: group A: Er:YAG laser, 2.94 microm/100 mJ/10 Hz/30 seconds; group B: diode laser, 810 nm/1.0 W/0.05 ms/30 seconds; group C: diode laser, 810 nm/1.4 W/0.05 ms/30 seconds. The temperature was monitored by means of a type T thermocouple (copper-constantan) positioned in the pulp chamber to assess pulpal temperature during and before irradiation. Afterwards, the specimens were longitudinally sectioned, and the buccal and lingual surfaces of each root were analyzed by scanning electron microscopy. RESULTS: In the Er:YAG laser group, the thermal analysis revealed an average temperature of -2.2 +/- 1.5 degrees C, while in the diode laser groups, temperatures were 1.6 +/- 0.8 degrees C at 1.0 W and 3.3 +/- 1.0 degrees C at 1.4 W. Electronic micrographs revealed that there were no significant morphological changes, such as charring, melting, or fusion, in any group, although the specimens were found to be more irregular in the Er:YAG laser group. CONCLUSIONS: The application of Er:YAG and diode lasers at the utilized parameters did not induce high pulpal temperatures. Root surface irregularities were more pronounced after irradiation with an Er:YAG laser than with a diode laser. PMID- 12886995 TI - Adhesion of Porphyromonas gingivalis serotypes to pocket epithelium. AB - BACKGROUND: Porphyromonas gingivalis, a key pathogen in periodontitis, is able to adhere to and invade the pocket epithelium. Different capsular antigens of P. gingivalis have been identified (K-serotyping). These P. gingivalis capsular types show differences in adhesion capacity to human cell lines or to cells cultured on a feeder layer or stromal equivalent. METHODS: The adhesion capacity of different P. gingivalis serotypes (6 capsular types and non-encapsulated strains) was compared on in vitro cultured epithelial monolayers from periodontal pockets of patients with periodontitis. The degree of adherence of P. gingivalis was evaluated by both culture and fluorescence microscopy. RESULTS: Non encapsulated strains adhered significantly more than their capsulated variants. Capsule type 4 (K-4) adhered slightly better than the remaining K-types. CONCLUSION: This study indicates that the presence and type of capsule have a significant influence on the initial adhesion of P. gingivalis to human periodontal pocket epithelial cells. PMID- 12886996 TI - Platelet-rich plasma contains high levels of platelet-derived growth factor and transforming growth factor-beta and modulates the proliferation of periodontally related cells in vitro. AB - BACKGROUND: Platelet-rich plasma (PRP) is a fraction of plasma, in which platelet derived growth factor (PDGF) and transforming growth factor-beta (TGF-beta) are thought to be concentrated. It is plausible that topically-applied PRP up regulates cellular activity and subsequently promotes periodontal regeneration in vivo. However, the concentrations of these growth factors in PRP have not been specifically determined and the biological effects of PRP at the cellular and molecular levels have not been determined. METHODS: PRP obtained from 20 healthy subjects was prepared from plasma by centrifugation. These PRP preparations were immediately subjected to an evaluation for PDGF-AB and TGF-beta1 using enzyme linked immunosorbent assay (ELISA) kits. The biological effects of the PRP preparations were evaluated on osteoblastic, epithelial, fibroblastic, and periodontal ligament cells. Cellular mitogenic activity was evaluated by counting cell numbers or evaluating 5-bromodeoxyuridine (BrdU) incorporation. Expression of alkaline phosphatase (ALP) was immunocytochemically evaluated. RESULTS: In the PRP preparations, platelets were concentrated up to 70.9 x 10(4) cells/microl (283.4% of the unconcentrated plasma). The levels of PDGF-AB and TGF-beta1 were also concentrated up to 182.0 ng/ml (440.6%) and 140.9 ng/ml (346.6%), respectively. Scatter plots revealed significant correlations between platelet counts and levels of these growth factors. PRP stimulated osteoblastic DNA synthesis and cell division (138% of control), with simultaneous down-regulation of ALP, but suppressed epithelial cell division (80% of control). PRP also stimulated DNA synthesis in gingival fibroblasts and periodontal ligament cells. CONCLUSIONS: These data demonstrated that both PDGF-AB and TGF-beta1 were highly concentrated in the PRP preparations. It is suggested PRP modulates cell proliferation in a cell type-specific manner similar to what has been observed with TGF-beta1. Since synchronized behavior of related cell types is thought to be required for successful periodontal regeneration, it is further suggested these cell type-specific actions may be beneficial for periodontal regenerative therapy. PMID- 12886997 TI - Platelet-rich plasma-derived fibrin clot formation stimulates collagen synthesis in periodontal ligament and osteoblastic cells in vitro. AB - BACKGROUND: Platelet-rich plasma (PRP) contains several growth factors, including platelet-derived growth factor (PDGF) and transforming growth factor-beta (TGF beta), at high levels. We have demonstrated the PRP functions like TGF-beta to modulate cell proliferation in a cell-type specific manner. In addition, PRP forms gel-like materials in several, but not all, cell cultures tested. This study was designed to investigate PRP's action on extracellular matrix production in periodontal ligament (PDL) and osteoblastic MG63 cell cultures. METHODS: PRP was prepared from the plasma obtained from autologous blood of healthy volunteers and stored at -20 degrees C until used. Cells treated with PRP (0.5% to 2%) were immunocytochemically stained for type I collagen and fibrin and the viscosity of the culture media was visually evaluated. Fibrinogen in PRP was detected by immunodot-blotting while endogenous thrombin expression in cells was detected by a modified enzyme-linked immunosorbent assay. RESULTS: Gel-like material rapidly (< 30 minutes) formed in cultures of either PDL or osteoblastic MG63 cell cultures after addition of PRP (> or = 0.5%). PRP changed cell shape and up regulated type I collagen at 24 hours. Fibrinogen was detected in the PRP preparations and insoluble fibrin networks were found in the newly formed gel like material. PRP's action on collagen synthesis was mimicked by purified fibrinogen and blocked by thrombin inhibitor. Thrombin was expressed both in PDL and MG63 cells. CONCLUSIONS: These findings demonstrated that the gel-like material formed in cell cultures of either PDL or MG63 cells is fibrin clot that is capable of up-regulating collagen synthesis in the extracellular matrix. Our data suggest the possibility that fibrinogen, converted to fibrin, in combination with growth factors present in PRP might effectively promote wound healing at sites of injury in periodontal tissue. PMID- 12886998 TI - Enhanced bone augmentation by controlled release of recombinant human bone morphogenetic protein-2 from bioabsorbable membranes. AB - BACKGROUND: The present study was undertaken to determine the effect of recombinant human bone morphogenetic protein-2 (rhBMP-2)-loaded biodegradable membranes on bone augmentation in a rabbit calvarial model. METHODS: Five microg of rhBMP-2 was loaded into a stiff hemispherical dome membrane made of poly(L lactide) and tricalcium phosphate (PLLA/TCP). The release kinetics of rhBMP-2 from the membrane were determined in vitro using a human BMP-2 immunoassay. Twelve rhBMP-2-loaded dome membranes (test group) and 12 control dome membranes (control group) were placed on the partial-thickness calvarial defects of 24 rabbits. The animals were sacrificed at 4 and 8 weeks, and undecalcified ground sections were prepared. Newly formed bone area and height were measured histomorphometrically and calculated by percentage ratio to the total submembranous space area and height below the dome. RESULTS: In vitro release results demonstrated that rhBMP-2 was released consistently over a 4-week period following a high initial burst release on the first day. At both 4 and 8 weeks, histomorphometric analysis revealed that the test group showed significantly higher newly formed bone heights and areas than the control group (P < 0.01). In the control group, new bone height was 36.3% of the dome height and the new bone area reached 8.2% of the submembranous space area at 8 weeks, while the test group reached 87.3% and 35.4%, respectively. CONCLUSION: These results suggest that the use of rhBMP-2-loaded PLLA/TCP membranes can result in additional bone augmentation, which is due to the osteoinductive properties of rhBMP-2 released from the membrane during healing. PMID- 12886999 TI - Hereditary gingival fibromatosis and expression of Ki-67 antigen: a case report. AB - BACKGROUND: Hereditary gingival fibromatosis (HGF) is a fibrotic enlargement of the gingiva. The mechanism that leads to the accumulation of abnormal amounts of gingival tissue in HGF is still unknown. The aim of this report was to present the clinical and histopathologic characteristics of a patient with gingival fibromatosis and to evaluate the proliferation of HGF fibroblasts. METHODS: We examined the proliferation rate of fibroblasts in this case by using Ki-67 immunohistochemical staining and compared the rate to fibroblasts of non fibromatosis gingival tissues from 5 healthy patients serving as controls. RESULTS: There were no Ki-67-positive cells in the lesional tissue, and the control gingiva revealed no immunostaining. The number of Ki-67 antigen-positive epithelial cell nuclei was observed to be low in the basal cell layers of hyperplastic gingival epithelia, similar to the control group. CONCLUSIONS: In the present case, there was no increase in the proliferation rate of lesional fibroblasts observed by Ki-67 immunohistochemical staining as a proliferation marker; only the epithelium was stained. It seems likely that the underlying mechanism of HGF may be an increase in the biosynthesis of collagen and glycosaminoglycans rather than cell proliferation. PMID- 12887000 TI - Two cases of linear IgA disease with clinical manifestations limited to the gingiva. AB - Linear IgA disease (LAD) is a chronic, subepithelial blistering disease that is associated with the presence of linear deposits of IgA along the basement membrane zone. Etiopathogenic aspects of LAD are only partially known. LAD is clinically characterized by vesiculobullous skin and mucous lesions. Although more than half of LAD patients present oral mucosal lesions, there are only a few cases reported of oral lesions as the only manifestation of LAD. This study presents 2 of these uncommon cases. PMID- 12887001 TI - Ameloblastoma of the mandible: a case report. AB - This report describes the case of a 60-year-old male who presented with a radiolucent lesion between his mandibular right premolars. The original radiographic diagnosis was a lateral periodontal cyst, but surgery produced biopsy material that proved to be an ameloblastoma. A discussion on developing a differential diagnosis is presented. PMID- 12887002 TI - Use of orthodontic treatment as an aid to third molar extraction: a method for prevention of mandibular nerve injury and improved periodontal status. AB - BACKGROUND: Impaction of mandibular third molars predisposes to pathological conditions including periodontal disease. Extraction of these teeth also may lead to damage to the nerve and to periodontal involvement of the second molars. This report describes a series of cases in which the third molars were orthodontically induced to erupt to prevent the sequelae associated with extraction. METHODS: Impacted mandibular third molars in 18 patients were surgically exposed following placement of an orthodontic appliance. Depending on the individual case, 1 of 3 approaches was used: attachment of a bracket, placement of a post in the root canal, or placement of an orthodontic wire through a bucco-lingual canal. After suturing the mucoperiosteal flap, the orthodontic appliance was activated. After the tooth erupted, it was removed and periodontal parameters were measured on the second molar. RESULTS: No damage to the inferior alveolar nerve was found. Probing depths on the second molar were reduced from 7.9 +/- 1.6 mm on the buccal and 7.4 +/- 1.0 mm on the lingual to 1.8 +/- 0.7 mm and 1.9 +/- 0. 7 mm, respectively. There was an average gain of 5.0 mm in attachment. Keratinized tissue increased from 2.9 +/- 0.7 to 3.8 +/- 0.6 mm. CONCLUSIONS: The interdisiplinary use of periodontics and orthodontics results in non-surgical removal of impacted mandibular third molars without damage to the inferior alveolar nerve and iatrogenic periodontal sequelae to the second molars. PMID- 12887003 TI - The subepithelial connective tissue graft with a vestibular releasing incision. AB - Initially, the rationale for mucogingival surgery was for function and health. In the past few years, esthetic results from augmentation procedures have become a goal. The subepithelial connective graft is a technique to achieve root coverage. Anatomically it is a procedure that cannot be utilized in every case. A difficult situation is the shallow vestibule with no keratinized gingiva and a strong frenulum attachment. The subepithelial connective tissue graft with a vestibular release as described here can achieve esthetic root coverage despite a shallow vestibule. PMID- 12887004 TI - Bioabsorbable membrane and bioactive glass in the treatment of intrabony defects in patients with generalized aggressive periodontitis: results of a 12-month clinical and radiological study. AB - BACKGROUND: Guided tissue regeneration can be achieved using membranes. In recent years, some evidence has been provided that bioactive glass is also capable of supporting the regenerative healing of periodontal lesions. The aim of this clinical and radiological prospective study was to compare the effectiveness of a bioabsorbable membrane and a bioactive glass in the treatment of intrabony defects in patients with generalized aggressive periodontitis. METHODS: Twelve patients (9 females, 3 males) with generalized aggressive periodontitis were enrolled in the study. The investigations were confined to 1- to 3-walled intrabony defects with a depth > or = 4 mm and with preoperative probing depths > or = 7 mm. Teeth with furcation involvement were excluded. Fifteen of the total 30 defects were treated with the membrane (RXT group) and 15 with the bioactive glass (PG group). Allocation to the groups was randomized. The clinical parameters plaque index (PI), gingival index (GI), probing depth (PD), bleeding on probing (BOP), gingival recession (GR), clinical attachment level (CAL), and mobility were recorded prior to surgery as well as 6 and 12 months after surgery. Intraoral radiographs were taken in standardized paralleling technique at baseline and 12 months after the surgery. Following digitization, changes in the distances from the alveolar crest (xCA) to the defect base (xBD) and from the CEJ (xCEJ) to the xCA were determined. Statistical analysis was based on Kolmogorov Smirnov test, Wilcoxon signed-ranks test, analysis of covariance, and Spearman's bivariate correlation analysis. RESULTS: After 12 months, a reduction in PD of 4.0 +/- 2.1 mm (P < 0.001) and a gain in CAL of 3.4 +/- 2.3 mm (P < 0.001) was registered in the RXT group. There was a slight increase in GR by 0.6 +/- 1.5 mm (P = 0.074). In the PG group, a reduction in PD of 3.8 +/- 1.9 mm (P < 0.001) and a gain in CAL of 2.8 +/- 1.9 mm (P < 0.001) was recorded, whereas GR increased by 1.0 +/- 1.4 mm (P = 0.007). The 6-month results did not differ significantly from the stated values. Radiographically, the defects (xCA to xBD) were found to be filled by 57.2 +/- 33.5% (P = 0.001) in the RXT group and by 50.5 +/- 22.8% (P = 0.001) in the PG group. Crestal resorption (xCEJ to xCA) was 6.1 +/- 34.5% (P = 0.910) in the RXT group and 15.1 +/- 39.7% (P = 0.433) in the PG group. Only the change in gingival recession after 12 months was significantly greater (P = 0.031) in the PG group, with -1.0 +/- 1.4 mm, compared to the RXT group, with 0.6 +/- 1.5 mm. Changes in the other clinical and radiological parameters showed no significant differences. The attachment gain correlated negatively with the preoperative PI (r = -0.574; P = 0.004) and with BOP after 6 months (r = -0.315; P = 0.021). CONCLUSIONS: Highly significant improvements in the parameters PD, CAL, and xCA-xBD were recorded after 6 and 12 months, respectively, with both regenerative materials. A good standard of oral hygiene and inflammation-free periodontal tissue in the postoperative phase improved the treatment outcome. PMID- 12887005 TI - Radiologic and histomorphometric evaluation of maxillary sinus grafting with alloplastic graft materials. AB - BACKGROUND: Sinus lifting procedures are widely used to obtain adequate bony support for implant placement at the atrophic maxillae. The aim of this study was to compare various sinus lifting and grafting techniques and materials. METHODS: Nine maxillae were treated with delayed and 46 maxillae with immediate implant placement techniques. A total of 104 implants were inserted. Panoramic radiographs were obtained prior to, after, and 6 to 8 months after surgery. Computed tomographies were also taken before and after surgery. The height of new bone was compared. Biopsy specimens were obtained during delayed implant placement and analyzed histomorphologically. RESULTS: There were no statistically significant differences between the panoramic radiographs for delayed and immediately placed implants, or between the graft materials. We observed correlations between the panoramic radiographs and computerized tomographies. CONCLUSION: Both delayed and immediate placement of implants can be used safely for sinus lifting. There were no statistically significant differences between the various graft materials. PMID- 12887006 TI - Meta-analysis of local tetracycline in treating chronic periodontitis. AB - BACKGROUND: Meta-analysis was used to assess the clinical efficacy of local delivery of tetracycline alone or as an adjunct to conventional mechanical therapy in patients with chronic periodontitis. METHODS: Studies were identified in MEDLINE and others sources. Meta-analyses were performed on the basis of probing depth (PD) at baseline, type of antimicrobial used, and experimental and control regimens (i.e., tetracycline plus scaling and root planing [SRP] versus SRP, tetracycline versus SRP, and tetracycline versus placebo, or no treatment). The effect of local tetracycline was evaluated for follow-up times of 4, 8, 12, 16, 24, and 36 weeks. Sensitivity analysis was performed according to antimicrobial delivery mode (irrigation, fibers, strips). A random effects model was used. RESULTS: The literature search identified 29 studies that met our inclusion criteria and were entered Into the meta-analysis. A significant mean reduction in PD for the combined tetracycline and SRP was observed regardless of initial probing depth and independently to the duration of follow-up. Tetracycline alone did not perform better than SRP, whereas they performed significantly better than placebo. Differences in improvement of attachment level (AL) were substantially similar to those encountered for PD. CONCLUSIONS: Our results documented that local. delivery of tetracycline improves the clinical outcomes of traditional treatment and should be considered particularly as an adjunct to SRP. Considerations regarding the adverse effects of widespread use of tetracycline should be taken into account when choosing a therapeutic strategy of chronic periodontitis. PMID- 12887007 TI - Tooth extraction durinig the course of periodontal therapy. PMID- 12887008 TI - The use of conscious sedation by periodontists. PMID- 12887009 TI - ATP synthases: insights into their motor functions from sequence and structural analyses. AB - ATP synthases are motor complexes comprised of F0 and F1 parts that couple the proton gradient across the membrane to the synthesis of ATP by rotary catalysis. Although a great deal of information has been accumulated regarding the structure and function of ATP synthases, their motor functions are not fully understood. For this reason, we performed the alignments and analyses of the protein sequences comprising the core of the ATP synthase motor complex, and examined carefully the locations of the conserved residues in the subunit structures of ATP synthases. A summary of the findings from this bioinformatic study is as follows. First, we found that four conserved regions in the sequence of gamma subunit are clustered into three patches in its structure. The interactions of these conserved patches with the alpha and beta subunits are likely to be critical for energy coupling and catalytic activity of the ATP synthase. Second, we located a four-residue cluster at the N-terminal domain of mitochondrial OSCP or bacterial (or chloroplast) delta subunit which may be critical for the binding of these subunits to F1. Third, from the localizations of conserved residues in the subunits comprising the rotors of ATP synthases, we suggest that the conserved interaction site at the interface of subunit c and delta (mitochondria) or epsilon (bacteria and chloroplasts) may be important for connecting the rotor of F1 to the rotor of F0. Finally, we found the sequence of mitochondrial subunit b to be highly conserved, significantly longer than bacterial subunit b, and to contain a shorter dimerization domain than that of the bacterial protein. It is suggested that the different properties of mitochondrial subunit b may be necessary for interaction with other proteins, e.g., the supernumerary subunits. PMID- 12887010 TI - New genes encoding subunits of a cytochrome bc1-analogous complex in the respiratory chain of the hyperthermoacidophilic crenarchaeon Sulfolobus acidocaldarius. AB - The soxL gene from Sulfolobus acidocaldarius (DSM 639) encodes a Rieske iron sulfur protein. In this study we report the identification of two open reading frames in its downstream region. The first one, named soxN, codes for a membrane protein bearing a resemblance to the b-type cytochromes of the cytochrome bc1 and b6f complexes. The protein is predicted to contain at least 10 transmembrane helices and features the two conserved histidine pairs coordinating the heme groups of these cytochromes. The second open reading frame, named odsN, encodes a soluble protein of unknown function. The genomic region displays a complex transcription pattern. Northern blot and RT-PCR analyses revealed the presence of mono- and bi-cistronic transcripts as well as a tri-cistronic transcript of soxL and cbsAB, encoding the mono-heme cytochrome b558/566. Phylogenetic analyses of the genes of the soxLN pair and of other archaeal gene pairs encoding Rieske iron sulfur proteins and b-type cytochromes revealed an identical branching patterns for both protein families, suggesting an evolutionary link of these genes provided by the functional interaction of the proteins. On the basis of the findings of this study and the previously studied properties of the soxL and cbsA proteins, we propose the occurrence of a novel cytochrome bc1-analogous complex in the membranes of Sulfolobus, consisting of the cytochrome b homolog soxN, the Rieske protein soxL, the high potential cytochrome cbsA, as well as the non-redox active subunits cbsB and odsN. PMID- 12887011 TI - Citrate uniport by the mitochondrial tricarboxylate carrier: a basis for a new hypothesis for the transport mechanism. AB - The tricarboxylate transport system located in the inner mitochondrial membrane was studied as an isolated protein reconstituted in proteoliposomes. The effects on the transport of citrate by various reagents, specific for different aminoacid residues, were analyzed. In the group of SH reagents, it was found that N ethylmaleimide is an irreversible inhibitor of the citrate-citrate exchange, while HgCl2 and the mercurial mersalyl cause a rapid unidirectional efflux of citrate from liposomes. It was demonstrated that NEM and mercurials act on different SH groups. Dithioerythritol is not able to reverse the effect of mersalyl unless another reagent, pyridoxalphosphate, is present. Pyridoxalphosphate itself, a reagent specific for NH2 residues, is an effective inhibitor of citrate exchange transport, as measured in both influx and efflux, but it has no effect on the mercurial-induced efflux. The same behavior was observed with diethylpyrocarbonate, a reagent specific for histidine and tyrosine residues. Interestingly, a slow basic efflux of internal citrate, in the absence of countersubstrate, was observed in proteoliposomes. Because it is inhibited by the same reagents acting on the exchange process, it is deduced that it is catalyzed by the tricarboxylate carrier. The ability of the carrier to perform a uniport of the substrate suggests the presence of a single substrate binding site on the carrier protein. A preliminary kinetic approach indicates that such a transport model is compatible with this theory. PMID- 12887012 TI - Adenine nucleotide translocase mediates the K(ATP)-channel-openers-induced proton and potassium flux to the mitochondrial matrix. AB - KATP channel openers have been shown to protect ischemic-reperfused myocardium by mimicking ischemic preconditioning, although their mechanisms of action have not been fully clarified. In this study we investigated the influence of the adenine nucleotide translocase (ANT) inhibitors--carboxyatractyloside (CAT) and bongkrekic acid (BA)--on the diazoxide- and pinacidil-induced uncoupling of isolated rat heart mitochondria respiring on pyruvate and malate (6 + 6 mM). We found that both CAT (1.3 microM) and BA (20 microM) markedly reduced the uncoupling of mitochondrial oxidative phosphorylation induced by the K(ATP) channel openers. Thus, the uncoupling effect of diazoxide and pinacidil is evident only when ANT is not fixed by inhibitors in neither the C- nor the M conformation. Moreover, the uncoupling effect of diazoxide and pinacidil was diminished in the presence of ADP or ATP, indicating a competition of K(ATP) channel openers with adenine nucleotides. CAT also abolished K+-dependent mitochondrial respiratory changes. Thus ANT could also be involved in the regulation of K(ATP)-channel-openers-induced K+ flux through the inner mitochondrial membrane. PMID- 12887013 TI - The composition of the incubation medium influences the sensitivity of mitochondrial permeability transition to cyclosporin A. AB - The aim of this work was to study permeability transition, and the influence of the composition of the incubation medium, on the inhibitory action of cyclosporin A. It was found that cyclosporin inhibited the opening of a nonspecific pore, as induced by the uncoupler carbonyl cyanide m-chlorophenylhydrazone, provided K+ was present in the incubation medium, but failed to do so if mitochondria are incubated in sucrose or Na+-based medium. It was also found that the sensitivity of mitochondria to the uncoupler depended on the incubation mixture, being more sensitive when sucrose was the osmotic support. Matrix Ca2+ release, large amplitude swelling, and drop in transmembrane electric gradient revealed permeability transition. The titration of membrane thiol groups shows them to be increased in mitochondria incubated in sucrose medium, in comparison with the values found in mitochondria incubated in KCl or NaCl medium. Our proposal is that the incubation in sucrose medium propitiated a conformational change of membrane proteins in such a way that cyclosporin was unable to bind to its target site. PMID- 12887014 TI - Energy conservation and dissipation in mitochondria isolated from developing tomato fruit of ethylene-defective mutants failing normal ripening: the effect of ethephon, a chemical precursor of ethylene. AB - Alternative oxidase (AOX) and uncoupling protein (UCP) are present simultaneously in tomato fruit mitochondria. In a previous work, it has been shown that protein expression and activity of these two energy-dissipating systems exhibit large variations during tomato fruit development and ripening on the vine. It has been suggested that AOX and UCP could be responsible for the respiration increase at the end of ripening and that the cytochrome pathway could be implicated in the climacteric respiratory burst before the onset of ripening. In this study, the use of tomato mutants that fail normal ripening because of deficiencies in ethylene perception or production as well as the treatment of one selected mutant with a chemical precursor of ethylene have revealed that the bioenergetics of tomato fruit development and ripening is under the control of this plant hormone. Indeed, the evolution pattern of bioenergetic features changes with the type of mutation and with the introduction of ethylene into an ethylene-synthesis deficient tomato fruit mutant during its induced ripening. PMID- 12887015 TI - Hydrogen peroxide- and cell-density-regulated expression of NADH-cytochrome b5 reductase in HeLa cells. AB - Environmental conditions regulate the expression of different antioxidant enzymes in cell culture. We have studied the effect of cell density and hydrogen peroxide on the expression of NADH-cytochrome b5 reductase in HeLa cells. Polypeptide levels of the NADH-cytochrome b5 reductase increased about three fold in confluent HeLa cells compared to sparse cells. Addition of H2O2 to HeLa cells altered expression levels of the NADH-cytochrome b5 reducatase in a concentration dependent way, being sparse cells more sensitive to H2O2 addition than confluent cells. The presence of pyruvate, a H2O2 scavenger, produced a significant increment (200%) in the levels of NADH-cytochrome b5 reductase in sparse cells, but less increase (25%) in confluent cells, suggesting that generation of endogenous H2O2 could repress NADH-cytochrome b5 reductase expression, particularly in sparse cultures. Accordingly, confluent HeLa cells showed significantly lower levels of reactive oxygen species than cells in sparse cultures. Addition of tert-butylhydroquinone, a compound which generates reactive oxygen species through redox cycling, also reduced expression of the NADH cytochrome b5 reductase. Increments in several antioxidant enzymes taking place during confluency could participate in the increase of NADH-cytochrome b5 reductase expression by reducing reactive oxygen species levels in cells. Overall, our results support that acute oxidative stress caused by H2O2 inhibits the expression levels of NADH-cytochrome b5 reductase, most likely due to inhibition of SP1 transcriptional activity. On the other hand, adaptation to H2O2 involved increased expression of the cytochrome b5 reductase, supporting the existence of additional regulatory mechanisms. PMID- 12887017 TI - Intriguing data on preservation of Japanese encephalitis virus RNA in dead mosquito tissues. PMID- 12887016 TI - Mitochondrial dysfunctions during aging: vitamin E deficiency or caloric restriction--two different ways of modulating stress. AB - Caloric restriction (CR), which has been demonstrated to offset the age associated accrual of oxidative injury, involves a reduction in calory intake while maintaining adequate nutrition, preserves the activities of antioxidant enzymes in postmitotic tissues, maintains organ function, opposes the development of spontaneous diseases, and prolongs maximum life span in laboratory rodents. It has been proposed that reductions in Reactive Oxygen Species (ROS) production and cellular oxidative injury are central to the positive effects of CR. In the present investigation we studied the effect of CR and of a vitamin E deprived diet on mitochondrial structure and features in the liver of rats during aging, in order to ascertain the extent of modifications induced by these experimental conditions. CR rats displayed structural and functional mitochondrial properties (fatty acid pattern, respiratory chain activities, antioxidant levels, and hydroperoxide contents) similar to those of younger rats whilst vitamin E deficient rats appeared older than their own age. The mitochondria of the former, together with those of young rats, possessed the lowest Coenzyme Q9, hydroperoxide, and cytochrome contents as well as a suitable fatty acid membrane composition. Our study confirms that CR is a valuable tool in limiting aging related free-radical damage also at mitochondrial liver level. PMID- 12887018 TI - Enzootic transmission of the agent of human granulocytic ehrlichiosis among cottontail rabbits. AB - To determine whether the agent of human granulocytic ehrlichiosis (HGE) (Anaplasma phagocytophilum) may be maintained in a parallel enzootic cycle between cottontail rabbits and their ticks, we sampled these hosts from a zoonotic site during five transmission seasons. Evidence of infection was sought by microscopy and polymerase chain reaction from rabbit blood or splenic tissue, and from ticks collected from rabbits or from vegetation. Approximately 27% of all rabbits sampled contained evidence of active infection, and 66% were seropositive. The vectorial capacity of Ixodes dentatus was demonstrated by xenodiagnosis studies; in addition, 2% of host-seeking nymphs were infected. Haemaphysalis leporispalustris was not a competent vector. Because of their propensity to densely inhabit peridomestic sites, and because I. dentatus may be transported by birds, a parallel cycle of transmission in cottontail rabbits would facilitate introduction and perpetuation of the agent of HGE. PMID- 12887019 TI - Sensitivity and specificity of ultrasound detection and risk factors for filarial associated hydroceles. AB - To better understand risk factors for hydrocele as a consequence of Wuchereria bancrofti infection, 342 men more than 15 years of age in an endemic area in Papua New Guinea were evaluated. Thirty-four subjects (9.9%) had hydrocele by physical examination. Ultrasound examination detected hydroceles in 57 men (16.7%). Compared with ultrasonography, the sensitivity of physical examination was 44.3%, the specificity was 98.2%, and the positive predictive value was 73.5%. Hydrocele was independently associated with age (odds ratio [OR] = 3.3, P < 0.01) and intensity of infection as determined by filarial antigenemia (OR = 2.3, P = 0.07). Dilation of spermatic cord lymphatics detectable by ultrasound did not correlate with hydrocele, but was associated with the presence of infection. These observations suggest that filarial pathology of the male genitalia is under-reported when evaluated by physical examination alone and that duration and intensity of infection are risk factors for hydrocele. PMID- 12887020 TI - Rapid assessment for lymphatic filariasis in central Nigeria: a comparison of the immunochromatographic card test and hydrocele rates in an area of high endemicity. AB - The rapid immunochromatographic card test (ICT) for Wuchereria bancrofti circulating filarial antigen is being used to map areas endemic for lymphatic filariasis. However, the ICT is expensive; thus, surveys based on this test must be relatively limited. Our study was conducted to determine if village-based hydrocele surveys could be used to supplement the ICT surveys in the mapping activities. We compared in 144 Nigerian villages the two assessment methods, ICT and examination for clinical hydrocele, in random samples of 30 adults selected using a procedure that obtained 15 younger males (reported age = 16-39 years old) and 15 older males (> or = 40 years), based on the assumption that hydrocele rates may be more prevalent in older age groups. The men were asked if they had scrotal swelling, then examined and tested by the ICT. We found a weakly positive correlation between village prevalence determined by the ICT and hydrocele (r = 0.041, P < 0.001). Only villages with hydrocele rates of 20% or greater were also consistently classified as having endemic filariasis by the ICT. There was no correlation between an individual's ICT positivity and clinical presence of hydrocele, and questioning about scrotal swelling was not predictive for presence of hydrocele. More research is needed to determine if community level hydrocele prevalence surveys can offer an economical and broadly applicable supplement to the ICT for determining the endemicity of filariasis. PMID- 12887021 TI - Placental and oral delivery of Schistosoma mansoni antigen from infected mothers to their newborns and children. AB - A 63-kD Schistosoma mansoni antigen was detected in 149 (86%) of 174 umbilical cord blood sera from infected women at delivery. Specific IgG antibodies to this antigen were also detected in 80% of cord blood sera. The 63-kD antigen showed the same molecular mass by Western blotting when isolated from cord blood, maternal blood, breast milk, and urine from women infected with S. mansoni. This antigen was detected in the urine of 25 infants born to infected mothers and followed for 18-24 months after delivery. It was also detected in some infants up to 21 days after parturition and then disappeared at 28 days, demonstrating the influence of breast-feeding on persistence of antigen in infants born to infected women. Thus, exposure to Schistosoma antigens and maternal antibodies to this organism may influence the developing immune responses to natural infection or vaccination of children born in endemic areas. PMID- 12887022 TI - Comparison of a polymerase chain reaction and the Kato-Katz technique for diagnosing infection with Schistosoma mansoni. AB - Fecal samples from 194 individuals living in an area of Brazil endemic for Schistosoma mansoni were analyzed by a polymerase chain reaction (PCR) and the Kato-Katz parasitologic examination. Statistical analysis of the results showed a kappa index of 0.8 between the two methods. The prevalence of infection was 30.9% in three fecal samples examined by the Kato-Katz method, but 38.1% in one fecal sample examined by the PCR technique. Repeated survey of discordant results showed that five (41.6%) of 12 parasitologically negative cases for which PCR gave positive results were misdiagnosed by Kato-Katz examinations. The PCR technique showed a sensitivity of 96.7% and a specificity of 88% when the parasitologic examination was used as the reference test. The efficacy of cure with praziquantel was 87.8% in three parasitologic stool examinations and 75.6% in one PCR survey. These results demonstrate that the PCR assay might be a valuable alternative for diagnosing Schistosoma infections. PMID- 12887023 TI - Dengue fever with acute acalculous cholecystitis. AB - Dengue fever (DF) with acute acalculous cholecystitis is rarely reported. To investigate the incidence, treatment, and prognosis of acute acalculous cholecystitis in DF patients, we retrospectively studied 10 patients with DF and acute acalculous cholecystitis. From October 2001 to July 2002, 131 patients were diagnosed with DF. Ten of 131 DF patients (7.63%) had complications of acute acalculous cholecystitis. Two patients underwent cholecystectomy and one underwent percutaneous transhepatic gallbladder drainage due to poor resolution of acute acalculous cholecystitis. We found acute acalculous cholecystitis in a small proportion of patients with DF. In our experience, closely monitoring vital signs to avoid shock and correct thrombocytopenia to avoid bleeding could be adequate for most patients. In some cases, surgical treatment may be needed for DF fever patients with complications of diffuse peritonitis. PMID- 12887024 TI - Central nervous system involvement in experimental infection with Leishmania (Leishmania) amazonensis. AB - We describe the pathologic alterations of the central nervous system (CNS) observed in experimental tegumentary leishmaniasis in BALB/c and Swiss mice. The mice were subcutaneously infected with 10(4) amastigotes of Leishmania (Leishmania) amazonensis. Animals were killed and brains were removed for histologic and immunocytochemical studies. Histologic examination showed that 66.6% of infected mice had a discrete hyperemia and inflammatory infiltrate in the meninges, composed of mononuclear cells and neutrophils with no detectable parasites. However, parasitized macrophages were detected in the cerebral parenchyma, as well as mast cells, lymphocytes, and polymorphonuclear cells. Necrosis in the cerebral parenchyma was also observed. Confocal fluorescence microscopy showed that CD8+ T lymphocytes are the major component of the inflammatory infiltrate in the CNS. In addition to these cells, CD4+, CD11b, and dendritic cells are present, in small numbers, in the inflammatory processes of the CNS. Thus, L. amazonensis is able to cross the blood-brain barrier and cause significant pathologic changes in the CNS. PMID- 12887025 TI - Antimicrobial resistance of bacterial pathogens associated with diarrheal patients in Indonesia. AB - The antimicrobial susceptibility patterns for 2,812 bacterial pathogens isolated from diarrheal patients admitted to hospitals in several provinces in the cities of Jakarta, Padang, Medan, Denpasar, Pontianak, Makassar, and Batam, Indonesia were analyzed from 1995 to 2001 to determine their changing trends in response to eight antibiotics: ampicillin, trimethoprim-sulfamethoxazole, chloramphenicol, tetracycline, cephalothin, ceftriaxone, norfloxacin, and ciprofloxacin. Vibrio cholerae O1 (37.1%) was the pathogen most frequently detected, followed by Shigella spp. (27.3%), Salmonella spp. (17.7%), V. parahaemolyticus (7.3%), Salmonella typhi (3.9%), Campylobacter jejuni (3.6%), V. cholerae non-O1 (2.4%), and Salmonella paratyphi A (0.7%). Of the 767 Shigella spp. isolated, 82.8% were S. flexneri, 15.0% were S. sonnei, and 2.2% were S. dysenteriae (2.2%). The re emergence of Shigella dysenteriae was noted in 1998, after an absence of 15 years. Shigella spp. were resistant to ampicillin, trimethoprim-sulfamethoxazole, chloramphenicol, and tetracycline. Salmonella typhi and Salmonella paratyphi A were susceptible to all antibiotics tested, while Salmonella spp. showed various resistance patterns according to species grouping. A small number of V. cholerae O1 were resistant to ampicillin, trimethoprim-sulfamethoxazole, chloramphenicol, and tetracycline; however, they were still sensitive to ceftriaxon, norfloxacin, and ciprofloxacin. Similar results were shown for V. cholerae non-O1. Campylobacter jejuni showed an increased frequency of resistance to ceftriaxone, norfloxacin, and ciprofloxacin, but was susceptible to erythromycin. This study shows that except for C. jejuni and V. parahaemolyticus, which appeared to be resistant to ciprofloxacin, the majority of the enteric pathogens tested were still susceptible to fluoroquinolones. PMID- 12887026 TI - Seroprevalence of Trypanosoma cruzi infection among school-age children in the endemic area of Guatemala. AB - In support of the National Program for Chagas Disease Control, we conducted a cross-sectional study to estimate the seroprevalence rate of Trypanosoma cruzi infection across the five Departments (Chiquimula, Jalapa, Zacapa, Jutiapa, and Santa Rosa) that are believed to comprise the entire principal endemic area in Guatemala. Also, so that the results could be used to identify areas of active transmission, we conducted the survey in school-aged children. We used an experimental enzyme-linked immunosorbent assay with blood samples obtained by finger prick to estimate the seroprevalence of T. cruzi. This assay has been previously tested and showed good sensitivity and specificity. Overall, the seropositivity rate for T. cruzi infection was 5.28% (235 of 4,450). Of 173 communities evaluated, 35 (20.23%) had a seropositive rate ranging from 10% to 45%. A number of parameters, including but not limited to living conditions, were examined for possible association with seropositivity. While there are several associations, the strongest association with seropositivity is living in a house with a thatch roof. The survey results will permit the Ministry of Health to stratify T. cruzi-endemic communities, enabling local health authorities to efficiently focus on vector control operations. PMID- 12887027 TI - Chronic Chagas' disease in rhesus monkeys (Macaca mulatta): evaluation of parasitemia, serology, electrocardiography, echocardiography, and radiology. AB - Severe chronic damage to the heart and gastrointestinal tract in patients with Chagas' disease are often observed 10-20 years after the acute phase. The course of long-lasting infection with the Colombian strain of Trypanosoma cruzi was studied in seven rhesus monkeys infected for 15-19 years. Subpatent parasitemia was detected in all studied animals, using hemoculture (two of seven), artificial xenodiagnosis (three of seven), and a polymerase chain reaction PCR (six of six). High titers of specific IgG antibody to T. cruzi persisted throughout the chronic phase of infection. Abnormal electrocardiographic (three of six) and echocardiographic (one of six) patterns detected in the T. cruzi-infected monkeys were possibly related to parasite-triggered myocardial damage. The results suggest that rhesus monkeys experimentally infected with T. cruzi, besides reproducing the acute phase of Chagas' disease, also develop chronic chagasic cardiomyopathy. PMID- 12887028 TI - Short report: variation in mitochondrial 12S and 16S ribosomal DNA sequences in natural populations of Triatoma infestans (Hemiptera: Reduviidae). AB - Mitochondrial DNA sequences of the 12S and 16S ribosomal RNA genes were analyzed in five natural populations of the Chagas' disease vector Triatoma infestans from Argentina. DNA sequence comparisons of 878 basepairs (12S plus 16S) revealed 13 haplotypes. A total of 10 private haplotypes were found in four of the populations analyzed, suggesting low current levels of genetic exchange. The levels of genetic differentiation between the population of Chancani (Cordoba) and other two of the populations analyzed indicated significant deviation from a pattern of unrestricted gene flow. The haplotypic diversity and the private haplotypes found in the geographically closest localities of Chancani and El Jardin (La Rioja) suggest that the reduction in the population size by insecticide treatment did not avoid the recovery of the populations apparently from survivors of the same area. PMID- 12887029 TI - Alteration of clinical outcome and histopathology of yellow fever virus infection in a hamster model by previous infection with heterologous flaviviruses. AB - Using a recently described hamster model of severe yellow fever (YF), we examined the hypothesis that prior infection with heterologous flaviviruses protects against severe or fatal YF. Hamsters were singly or sequentially infected with Japanese encephalitis, St. Louis encephalitis, West Nile, and/or dengue-1 viruses, and then challenged with a virulent strain of yellow fever virus (YFV). In contrast to control (naive) hamsters, many of which appeared clinically ill or died after YFV infection, the flavivirus-immune animals remained asymptomatic. The flavivirus-immune hamsters also had a reduced viremia and lower serum levels of alanine aminotransferase and total bilirubin, compared with naive hamsters, following YFV infection. Histologically, livers of animals in the flavivirus immune and control groups showed comparable levels of multifocal necrapoptosis. However, steatosis was not observed in the flavivirus-immune animals, whereas naive hamsters developed extensive microvesicular steatosis in the liver following YFV infection. These findings suggest that hepatocytic steatosis is an adverse microscopic feature associated with severe disease in YFV infection. Our experimental results support earlier anecdotal reports that prior exposure of humans to heterologous flaviviruses reduces subsequent risk of fatal YFV infection. PMID- 12887030 TI - Acute undifferentiated fever caused by infection with Japanese encephalitis virus. AB - To determine the proportion of acute undifferentiated fevers without neurologic deficits related to infection with Japanese encephalitis (JE) virus, flavivirus serology (dengue and JE) was performed in a cohort of 156 adults presenting to a hospital in Chiangrai, Thailand. Recent flavivirus infection was diagnosed for any individual with an IgM result > 40 units. A ratio of dengue virus IgM to JE virus IgM < 0.91 defined a JE virus infection. Diagnostic criteria for Japanese encephalitis were met in 22 individuals (14%), and were unequivocal in 8 patients. The admission findings in these eight subjects were similar to those described for other flavivirus infections. Thrombocytopenia was the most striking laboratory abnormality (median platelet count = 119,000/mm3, range = 44,000 236,000/mm3). Headache (75%), nausea (50%), myalgia (38%), rash (38%), and diarrhea (25%) were the most frequently encountered signs and symptoms. Infection with Japanese encephalitis virus is an underappreciated cause of acute undifferentiated fever in Asia. PMID- 12887031 TI - Case report: gnathostomiasis in two travelers to Zambia. AB - Gnathostomiasis is a systemic infection caused by migrating nematode larvae of the genus Gnathostoma. It is a zoonosis involving a wide variety of animals as intermediate and definitive hosts, and consumption of raw fish is the main risk factor. The condition is most commonly seen in southeastern Asia, but has been described in a number of other countries, all outside Africa. We report the infection in two travelers returning from southcentral Africa, who presented with non-specific symptoms and marked eosinophilia, and in whom schistosomiasis was initially suspected. The typical migratory skin lesions of gnathostomiasis appeared later. The infections responded well to albendazole. The patients acquired the infection in western Zambia; this region of Africa appears to be a newly identified risk area for gnathostomiasis in tourists who indulge in eating raw freshwater fish. PMID- 12887032 TI - Cryptosporidium parvum in children with diarrhea in Mulago Hospital, Kampala, Uganda. AB - A cross-sectional case-control study (ratio = 3:1) was conducted over a 15-month period to determine the prevalence and consequences of cryptosporidiosis in hospitalized diarrheic children (0-5 years old) at Mulago Hospital in Kampala, Uganda. Cryptosporidium parvum was detected and genotyped among 2,446 children of whom 1,779 (72.7%) had diarrhea, and 667 (27.3%) were age- and sex-matched controls. Of the 1,779 children with diarrhea, 532 (29.9%) had persistent (> 14 days) diarrhea and 1,247 (70.1%) had acute diarrhea. Overall, 444 (25.0%) of the 1,779 children with diarrhea had C. parvum, compared with only 57 (8.5%) of the 667 children without diarrhea (chi2 = 80.2, P < or = 0.0001). Within this group of infected children, 72.8% were infected with genotype 1, 18.4% with genotype 2, and 4.1% with a mixture of both genotypes, and 4.1% isolates were either unclassified or C. meleagridis. The prevalence was highest during the rainy months of April to June. Of the 532 children with persistent diarrhea, 166 (31.2%) had C. parvum compared with 278 (22.3%) of the 1,247 children with acute diarrhea (chi2 = 15.8, P < or = 0.0001). There was a significant association between C. parvum and malnutrition including stunting, being underweight, and wasting. Unfavorable outcome (death or failure to resolve within 14 days) occurred in 139 (72.8%) of the 191 children with C. parvum, and in only 65.1% of the 545 without (odds ratio = 1.117, 95% confidence interval = 1.005-1.243, P = 0.05), Of the 191 children with C. parvum, 24 (12.6%) died, compared with 34 (6.2%) of the 545 without C. parvum (P = 0.005). Mortality rates were higher among children with severe dehydration and persistent diarrhea, and in stunted or underweight children infected with C. parvum. Among Ugandan children, cryptosporidiosis, which remains untreatable, is frequently associated with diarrhea and other serious and unfavorable consequences. PMID- 12887033 TI - Sexual transmission of hepatitis B virus, hepatitis C virus, and human immunodeficiency virus type 1 infections among male transvestite comercial sex workers in Montevideo, Uruguay. AB - Prostitution may constitute a risk behavior for infection with hepatitis B virus (HBV), hepatitis C virus (HCV), and human immunodeficiency virus (HIV). We conducted a seroepidemiologic study among 200 male transvestite commercial sex workers (CSWs) in Montevideo, Uruguay in 1999. Evidence of exposure to HBV, HCV, and HIV was found in 101 (50.5%), 13 (6.5%), and 43 (21.5%) individuals, respectively. Positivity for HIV was correlated with both HBV (odds ratio [OR] = 2.15, 95% confidence interval [CI] = 1.01-4.67) and HCV (OR = 3.47, 95% CI = 0.90 12.79) infection. Predictors of infection were older age (> or = 26 years; P < 0.01) for all 3 viruses and time in CSW (> or = 5 years; P < 0.05) for HBV and HIV. Prior history of use of drugs (OR = 3.54, 95% CI = 1.09-11.52) and sexual contact with foreigners (OR = 9.2, 95% CI = 1.16-73.12) were found to be associated only with HCV infection. Sexual transmission of these viruses constitutes a significant problem among male transvestite CSWs. PMID- 12887034 TI - Low prevalence and increased household clustering of Mycobacterium tuberculosis infection in high altitude villages in Peru. AB - Anecdotal historical evidence suggests that tuberculosis was uncommon at high altitude, but whether transmission is affected by high altitude is not known. To test whether high altitude lowers infection by Mycobacterium tuberculosis, the prevalence of tuberculin skin test (TST) positivity was compared between two high altitude villages (3,340 meters [10,960 feet] and 3,500 meters [11,480 feet]) and three sea-level sites in Peru. High altitude villages had lower TST-positive prevalence rates (5.7% and 6.8%) than sea level areas (25-33%), and the difference remained significant (odds ratio = 4.5-6.0) after adjusting for age, education, bacille Calmette-Guerin vaccination, and contact with tuberculosis patients. The TST-positive individuals clustered within highland families more than within sea level families. These data suggest that prevention and control efforts targeted to families may be more effective at high altitude. The mechanism by which TST-positivity prevalence is decreased at high altitude is unknown, but may reflect relative hypoxia, low humidity, or an increased ultraviolet effect. PMID- 12887035 TI - Antibody-mediated in vitro growth inhibition of field isolates of Plasmodium falciparum from asymptomatic children in Burkina Faso. AB - Antibody-mediated inhibition of Plasmodium falciparum parasites in vitro reflects the potential parasite-neutralizing activity of the antibodies in vivo. In this study, immunoglobulins and P. falciparum isolates were collected from children with asymptomatic malaria in Burkina Faso. We demonstrate a significantly lower in vitro growth inhibitory activity against the P. falciparum field isolates by autologous host immunoglobulin compared with that of immunoglobulin from other individuals. To gain further insight to possible mechanisms for the diverse sensitivity observed, analyses of consecutive isolates taken 14 days apart were performed with regard to polymerase chain reaction-based genotyping and sensitivity to growth inhibition in vitro. All the asymptomatic infections were composed of multiple, genotypically distinct parasite clones, and at least one new parasite clone appeared in most of the day 14 isolates compared with the corresponding day 0 isolates. Apparently persisting parasite clones, present in both the day 0 and day 14 isolates from the same person, were also frequently observed. The day 14 isolates were more effectively inhibited by autologous day 14 immunoglobulin than by the corresponding day 0 immunoglobulin in 57% of the cases. However, the frequent presence of persisting parasite clones in asymptomatic children indicates that the parasite may develop a relative resistance to neutralizing immune responses. PMID- 12887036 TI - Spatial and temporal heterogeneity of Anopheles mosquitoes and Plasmodium falciparum transmission along the Kenyan coast. AB - The seasonal dynamics and spatial distributions of Anopheles mosquitoes and Plasmodium falciparum parasites were studied for one year at 30 villages in Malindi, Kilifi, and Kwale Districts along the coast of Kenya. Anopheline mosquitoes were sampled inside houses at each site once every two months and malaria parasite prevalence in local school children was determined at the end of the entomologic survey. A total of 5,476 Anopheles gambiae s.l. and 3,461 An. funestus were collected. Species in the An. gambiae complex, identified by a polymerase chain reaction, included 81.9% An. gambiae s.s., 12.8% An. arabiensis, and 5.3% An. merus. Anopheles gambiae s.s. contributed most to the transmission of P. falciparum along the coast as a whole, while An. funestus accounted for more than 50% of all transmission in Kwale District. Large spatial heterogeneity of transmission intensity (< 1 up to 120 infective bites per person per year) resulted in correspondingly large and significantly related variations in parasite prevalence (range = 38-83%). Thirty-two percent of the sites (7 of 22 sites) with malaria prevalences ranging from 38% to 70% had annual entomologic inoculation rates (EIR) less than five infective bites per person per year. Anopheles gambiae s.l. and An. funestus densities in Kwale were not significantly influenced by rainfall. However, both were positively correlated with rainfall one and three months previously in Malindi and Kilifi Districts, respectively. These unexpected variations in the relationship between mosquito populations and rainfall suggest environmental heterogeneity in the predominant aquatic habitats in each district. One important conclusion is that the highly non-linear relationship between EIRs and prevalence indicates that the consistent pattern of high prevalence might be governed by substantial variation in transmission intensity measured by entomologic surveys. The field-based estimate of entomologic parameters on a district level does not provide a sensitive indicator of transmission intensity in this study. PMID- 12887037 TI - Efficacy of amodiaquine alone and combined with sulfadoxine-pyrimethamine and of sulfadoxine pyrimethamine combined with artesunate. AB - The safety and the efficacy of amodiaquine (AQ) alone, AQ plus sulfadoxine pyrimethamine (SP) (AQ plus SP), and artesunate (ART) plus SP (ART plus SP), three possible alternatives to chloroquine (CQ), were investigated in 379 Rwandan children 6-59 months old with uncomplicated Plasmodium falciparum malaria who visited one urban/peri-urban health center and two rural health centers. The three treatment regimens were well tolerated and no serious adverse effects were observed. Children treated with AQ plus SP had less clinical failures than those treated with ART plus SP (odds ratio [OR] = 0.25, 95% confidence interval [CI] = 0.06-0.81, P = 0.01) or AQ alone (OR = 0.33, 95% CI = 0.07-1.10, P = 0.08). Even after new infections were excluded, AQ plus SP was still significantly more efficacious than ART plus SP (P = 0.05). At day 14, the mean packed cell volume was significantly higher in the AQ plus SP group compared with the ART plus SP group (P = 0.02) and with the AQ alone group (P = 0.01). In Rwanda, AQ plus SP has been chosen to replace CQ as a first-line treatment. However, this is considered an interim measure and new combinations, possibly co-formulated, should be identified and tested. PMID- 12887038 TI - Enhancement of development of larval Anopheles arabiensis by proximity to flowering maize (Zea mays) in turbid water and when crowded. AB - To determine whether proximity to flowering maize enhances the development of larval anopheline mosquitoes breeding in turbid water and when crowded, we evaluated the development of larval Anopheles arabiensis under various conditions of turbidity, larval density, and proximity to pollen-shedding maize in simulated breeding puddles in a malaria-endemic site. In naturally formed puddles, water turbidity, as well as larval density, increased as the rainy season progressed. In sites remote from flowering maize, more pupae developed and the resulting adults were larger in relatively clear water than in turbid water, and larval crowding inhibited development. In close proximity to flowering maize, however, larval development was little affected by water turbidity and larval crowding. Larvae of this member of the African An. gambiae complex of mosquitoes develop readily in turbid water and when crowded, provided that their breeding sites are located where maize pollen is abundant. PMID- 12887040 TI - Alzheimer's disease as a disconnection syndrome? AB - This paper reviews the growing amount of evidence supporting the hypothesis that Alzheimer's disease includes a disconnection syndrome. This evidence came mainly from neuropathological, electrophysiological, and neuroimaging studies. Moreover, a few recent neuropsychological studies have also explored the effects of a disconnection between cerebral areas on cognitive functioning. Finally, and more generally, the contribution of this interpretation to the understanding of Alzheimer's disease cognitive deficits is considered. PMID- 12887041 TI - In search of a conceptualization of multiple sclerosis: a historical perspective. AB - A thorough understanding of Multiple Sclerosis (MS) is necessary to offer individuals informed options for treatment and planning. To assist in this quest, the following historical analysis examined how MS has been conceived from the 14th century through the early 20th century. Primary sources were consulted whenever possible, and many of the original archival materials were accessed by the first author (MB) during an on-site visit to the Rare Book Room of the New York Academy of Medicine. There is a striking similarity between how MS symptoms have presented throughout history compared with the 21st century. Sensorimotor and cognitive sequelae have been observed in patients since the 1800s. Cognitive symptoms were acknowledged in the 1800s, but disregarded in the early 1900s and were not given recognition again until the latter part of the 20th century. If conceptualizations of MS are inaccurate, patients will not be served well. In contrast to the shared symptomatology across time, early conceptualizations of etiology and treatment choices differed dramatically from today, a genuine reflection of the times in which they were created. PMID- 12887042 TI - Psychiatry in France. PMID- 12887039 TI - Verbal declarative memory dysfunction in schizophrenia: from clinical assessment to genetics and brain mechanisms. AB - The recent literature on the neuropsychology of schizophrenia has emphasized memory deficits as a key area of impairment. Abnormalities in the medial temporal lobe, a brain region crucial for long-term memory formation, have also consistently been reported. We conducted a comprehensive review of verbal declarative memory (VDM) in schizophrenia with the aim of systematically addressing the nature of this impairment. We conclude that verbal declarative memory is significantly impaired in schizophrenia and is largely accounted for by deficits in the encoding stage. Subtle impairments in increased rates of forgetting are present, but are mild compared with those in amnestic disorders. Impairment in other cognitive domains studied thus far (e.g., attention), medication effects, or fluctuations in symptoms do not completely account for the deficit. VDM is among the most impaired neurocognitive domains in schizophrenia (along with attention and executive functions). Milder encoding deficits are present in high-risk subjects and non-psychotic relatives of individuals with schizophrenia suggesting that components of the deficit are associated with a genetic vulnerability to the illness, and are independent of the frank psychotic illness. Furthermore, VDM is observed in individuals experiencing their first psychotic episode and it remains fairly consistent over time. Preliminary imaging studies and other work suggest abnormalities in prefrontal-hippocampal processing networks. Future work should emphasize delineating specific information processing components contributing to the deficit. This would allow imaging studies to determine which brain regions contribute to specific information processing deficits in schizophrenia. PMID- 12887043 TI - Psychiatric morbidity among housemaids in Kuwait. III: Vulnerability factors. AB - BACKGROUND: Housemaids are a relatively homogenous immigrant subgroup in terms of their gender; ethnic origin; and socio-cultural, educational and occupational background. Psychiatric morbidity among housemaids is two to five times higher than the native female population. AIMS: To determine the possible pre immigration risk factors for prospective psychiatric breakdown among the housemaids. METHODS: The sample consisted of all the housemaids (N = 197) hospitalised during the two-year study period. The controls comprised all the newly arrived housemaids (N = 502). The measures obtained included demographic characteristics and previous history of physical illness, psychiatric illness, hospitalisation and family history of psychiatric disorder. RESULTS: More than a quarter of the hospitalised group broke down within one month of their arrival. The hospitalised group had a significant excess of Sri Lankan housemaids; non Muslims; those with less than four years of education and those with a previous history of physical illness, psychiatric illness or hospitalisation. CONCLUSIONS: A number of potential risk factors results in premature repatriation of housemaids on mental health grounds. Preventive measures involving recruitment procedures and pre-departure orientation courses are needed to minimise the expatriate failure among the housemaids. PMID- 12887044 TI - Traditional Chinese religious beliefs and superstitions in delusions and hallucinations of Chinese schizophrenic patients. AB - Religious beliefs and superstitions have an important impact on the psychopathology of psychiatric patients. Traditional Chinese religious beliefs and superstitions, such as fortune telling, Buddhist gods, Taoist gods, historical heroic gods and ancestor worship, have important influence on subjective psychotic experiences, in particular delusions and hallucinations. By means of empirical phenomenological case narration, the writer shows that all these traditional Chinese religious beliefs and superstitions tend to affect the contents, manifestation and meaningfulness of delusion and hallucination. They also serve as a means to replace clients' self-identity. They appear in the form of a supernatural force to resolve all difficulties, cause of troubles and misfortune, stress and coping mechanisms. PMID- 12887045 TI - Violence, self-harm, victimisation and homelessness in patients admitted to an acute inpatient unit in South Australia. AB - OBJECTIVES: To measure the incidence of traumatic and potentially socially disruptive events prior to admission to a psychiatric hospital. METHOD: One hundred and nineteen patients were interviewed and further information was obtained from case notes. Data were collected concerning rates of violence, self harm, threats of violence or self-harm, physical assault and homelessness. RESULTS: The patients' mean age was 35 years, 70% were male and 77% were currently single. More than half were legally detained. The most common diagnoses were substance abuse disorders, schizophrenia and related psychoses and mood disorders. Eighteen per cent of patients were physically violent at the time of admission. They were more likely to have a diagnosis of substance abuse. Significantly more men (30%) than women (8%) made threats of violence. Eighteen per cent of patients had physically harmed themselves. Significantly more women (70%) than men (40%) had threatened to harm themselves. Eight per cent of patients had been physically assaulted prior to admission. Twenty-seven per cent of patients were homeless. CONCLUSION: These patients were severely disabled with high rates of comorbidity. Socially disruptive behaviours were common, and almost a quarter of the patients were homeless. This study highlights the need for development of targeted services in the community. PMID- 12887046 TI - Common mental disorders among primary care attenders in Vellore, South India: nature, prevalence and risk factors. AB - OBJECTIVE: The study examined the nature and prevalence of and the factors associated with common mental disorders in primary care in Vellore, South India. METHOD: Consecutive attenders (N = 327) to a primary health care facility were recruited. RESULTS: One hundred and eleven (33.9%) subjects satisfied criteria for common mental disorders. Depression was the common presentation (83.8%). Indicators of low socio-economic status (being in debt, inability to buy food, having less than three square meals per day) and illiteracy were significantly associated with caseness. CONCLUSIONS: Subjects with common mental disorders are commonly seen in primary health care settings. Illiteracy and poverty are associated with caseness. Primary health care research and policy needs to focus on common mental disorders. PMID- 12887047 TI - Emotional and somatic distress in eastern North Carolina: help-seeking behaviors. AB - Socio-demographic and cultural factors have been reported to shape help-seeking behavior. However, not much effort has been made to determine the effects of these factors on help-seeking among rural populations. A telephone survey using random-digit dialing was used to explore socio-demographic characteristics and ethnic differences in the types of professionals sought for unexplained somatic and emotional problems (N = 1161) in rural eastern North Carolina. Ethnic differences in comfort with participating in support groups were also examined. The effect of a large natural disaster, Hurricane Floyd and subsequent flooding, on help-seeking choices and comfort with support groups was also assessed. Results showed that the rural population makes a sharp distinction between somatic symptoms and stress-related symptoms. This distinction seemed more pronounced for European-Americans than for African-Americans. In general African Americans selected help-seeking from clergy more often than European-Americans, although for unexplained somatic symptoms this difference was fostered by Hurricane Floyd with its flooding. African-Americans showed markedly increased comfort with support groups after the hurricane, while European-Americans showed no changes in comfort with support groups as a function of the hurricane. The effects of Hurricane Floyd on African-Americans are interpreted as reflecting an increased salience of community support for African-Americans, significantly through the Baptist Church. Training of clergy should include recognition of stress-related somatic and emotional symptoms and the potential for an important referral role, especially following disasters. PMID- 12887048 TI - From whence comes mental illness stigma? AB - BACKGROUND: This paper seeks to answer two fundamental questions: What is the basis of the current form of mental illness stigma? and Why do western cultures stereotype people with mental illness as dangerous, incompetent and blameful, rather than something else? MATERIAL AND DISCUSSION: We argue that a motivational model called system-justification offers several benefits for answering these questions. System-justification portrays stigma as a way of making sense of economic and political differences between the majority and stigmatized subgroups. We contrast system-justification with two cognitive models of stigma that seem to have strong support from naive psychology: mental illness stigma results as the normal perception of a group of people who are dangerous and/or blameworthy and there is a kernel of truth to the stigmatizing attitudes about people with mental illness. Although research supporting the latter two models is mixed, there are significant limitations to the models, as well as concerns that normal perception and kernel of truth might actually promote stigma. CONCLUSIONS: As an alternative, system-justification combines three paradigms that suggest its worthiness for future research: 1) a review of historical and economic forces that influence social phenomena; 2) the need of humankind to understand these forces and organize them into a unitary framework; and 3) the cognitive mechanisms that are essential for this comprehension. Implications of this model for stigma change are discussed. PMID- 12887049 TI - Vito Turk--30 years of research on cysteine proteases and their inhibitors. PMID- 12887050 TI - Family C1 cysteine proteases: biological diversity or redundancy? AB - Recent progress in the identification and partial characterization of novel genes encoding cysteine proteases of the papain family has considerably increased our knowledge of this family of enzymes. Kinetic data available to date for this large family indicate relatively broad, overlapping specificities for most enzymes, thus inspiring a growing conviction that they may exhibit functional redundancy. This is also supported in part by phenotypes of cathepsin knockout mice and suggests that several proteases can substitute for each other to degrade or process a given substrate. On the other hand, specific functions of one particular protease have also been documented. In addition, differences in cellular distribution and intracellular localization may contribute to defining specific functional roles for some of these proteases. PMID- 12887051 TI - Molecular regulation of human cathepsin B: implication in pathologies. AB - Cathepsin B is a papain-family cysteine protease that is normally located in lysosomes, where it is involved in the turnover of proteins and plays various roles in maintaining the normal metabolism of cells. This protease has been implicated in pathological conditions, e.g., tumor progression and arthritis. In disease conditions, increases in the expression of cathepsin B occur at both the gene and protein levels. At the gene level, the altered expression results from gene amplification, elevated transcription, use of alternative promoters and alternative splicing. These molecular changes lead to increased cathepsin B protein levels and in turn redistribution, secretion and increased activity. Here we focus on the molecular regulation of cathepsin B and attendant implications for tumor progression and arthritis. The potential of cathepsin B as a therapeutic target is also discussed. PMID- 12887052 TI - Caspases and neuronal development. AB - Recent developments have shown that inappropriate activation of apoptotic pathways contributes to many neurodegenerative diseases. The basic mechanisms that underlie apoptosis in neurodegenerative diseases are uncertain, although they likely represent the subversion of normal developmental programs. Several types of neuronal cell death have been reported, including autophagic and caspase independent cell death. In this review we consider evidence for the participation of apoptotic caspases in neuronal development, and examine the hypothesis that differentiating neurons undergo stage-specific alterations in apoptosis sensitivity that may be due to caspase regulation. In addition, we present data supporting this hypothesis. PMID- 12887053 TI - Structural basis of the matrix metalloproteinases and their physiological inhibitors, the tissue inhibitors of metalloproteinases. AB - The matrix metalloproteinases (MMPs) constitute a family of multidomain zinc endopeptidases with a metzincin-like catalytic domain, which are involved in extracellular matrix degradation but also in a number of other important biological processes. Under healthy conditions, their proteolytic activity is precisely regulated by their main endogenous protein inhibitors, the tissue inhibitors of metalloproteinases. Disruption of this balance results in pathophysiological processes such as arthritis, tumor growth and metastasis, rendering the MMPs attractive targets for inhibition therapy. Knowledge of their tertiary structures is crucial for a full understanding of their functional properties and for rational drug design. Since the first appearance of atomic MMP structures in 1994, a large amount of structural information has become available on the catalytic domains of MMPs and their substrate specificity, interaction with synthetic inhibitors and the TIMPs, the domain organization, and on complex formation with other proteins. This review will outline our current structural knowledge of the MMPs and the TIMPs. PMID- 12887054 TI - Managing peptidases in the genomic era. AB - The enzymes that hydrolyse peptide bonds, called peptidases or proteases, are very important to mankind and are also very numerous. The many scientists working on these enzymes are rapidly acquiring new data, and they need good methods to store it and retrieve it. The storage and retrieval require effective systems of classification and nomenclature, and it is the design and implementation of these that we mean by 'managing' peptidases. Ten years ago Rawlings and Barrett proposed the first comprehensive system for the classification of peptidases, which included a set of simple names for the families. In the present article we describe how the system has developed since then. The peptidase classification has now been adopted for use by many other databases, and provides the structure around which the MEROPS protease database (http://merops.sanger.ac.uk) is built. PMID- 12887055 TI - Insights into the roles of cathepsins in antigen processing and presentation revealed by specific inhibitors. AB - Eleven human cathepsins have been identified, however, the in vivo roles of individual cathepsins are still largely unknown. In this brief review we will summarize the functions of individual cathepsins in antigen processing and presentation, which are the initial steps of the immune response. Two general inhibitors of papain-like cysteine proteases, E-64 and pyridoxal phosphate, can completely suppress antigen presentation in vivo. To evaluate the contribution of individual cathepsins, specific inhibitors have been developed based on cathepsin tertiary structures: CA-074 for cathepsin B, CLIK-148 and -195 for cathepsin L, CLIK-60 for cathepsin S. Administration of CA-074, a cathepsin B inhibitor, suppresses the response to exogenous antigens, such as hepatitis B virus antigen, ovalbumin and Leishmania major antigen, and induces switching of the helper T cell responses from Th-2 to Th-1 of CD4+ T cells, thereby downregulating the production of IgE and IgG1. Administration of the cathepsin S inhibitor CLIK-60 impairs presentation of an autoantigen, alpha-fodrin, in Sjogren's syndrome and suppresses the Th-1 response and autoantibody production. PMID- 12887056 TI - Cleavage site specificity of cathepsin K toward cartilage proteoglycans and protease complex formation. AB - Cathepsin K is a potent extracellular matrix-degrading protease that requires interactions with soluble glycosaminolycans for its collagenolytic activity in bone and cartilage. The major sources of glycosaminoglycans in cartilage are aggrecan aggregates. Therefore, we investigated whether cathepsin K activity is capable to hydrolyze aggrecan into fragments allowing the formation of glycosaminoglycan-cathepsin K complexes and determined the cleavage site specificity of cathepsin K toward the cartilage-resident link protein and aggrecan. The cleavage site specificity was compared with those of cathepsins S and L. All three cathepsins released glycosaminoglycans from native bovine cartilage at lysosomal pH and to a lesser degree at neutral extracellular pH. Cathepsin-predigested aggrecan complexes and cartilage provided suitable glycosaminoglycan fragments that allowed the formation of collagenolytically active cathepsin K complexes. A detailed analysis of the degradation of aggrecan aggregates revealed two cathepsin K cleavage sites in the link protein and several sites in aggrecan, including one site within the interglobular domain E1. In summary, these results demonstrate that cathepsin K is capable to degrade aggrecan complexes at specific cleavage sites and that cathepsin K activity alone is sufficient to self-provide the glycosaminoglycan fragments required for the formation of its collagenolytically active complex. PMID- 12887057 TI - Toward computer-based cleavage site prediction of cysteine endopeptidases. AB - Identification of relevant substrates is essential for elucidation of in vivo functions of peptidases. The recent availability of the complete genome sequences of many eukaryotic organisms holds the promise of identifying specific peptidase substrates by systematic proteome analyses in combination with computer-based screening of genome databases. Currently available proteomics and bioinformatics tools are not sufficient for reliable endopeptidase substrate predictions. To address these shortcomings the bioinformatics tool 'PEPS' (Prediction of Endopeptidase Substrates) has been developed and is presented here. PEPS uses individual rule-based endopeptidase cleavage site scoring matrices (CSSM). The efficiency of PEPS in predicting putative caspase 3, cathepsin B and cathepsin L cleavage sites is demonstrated in comparison to established algorithms. Mortalin, a member of the heat shock protein family HSP70, was identified by PEPS as a putative cathepsin L substrate. Comparative proteome analyses of cathepsin L deficient and wild-type mouse fibroblasts showed that mortalin is enriched in the absence of cathepsin L. These results indicate that CSSM/PEPS can correctly predict relevant peptidase substrates. PMID- 12887058 TI - Isolation and characterization of a novel and potent inhibitor of Arg-gingipain from Streptomyces sp. strain FA-70. AB - Arg-gingipain (Rgp) is a major cysteine proteinase produced by the oral bacterium Porphyromonas gingivalis, which is a major pathogen of advanced periodontal diseases. This enzyme is important for the bacterium both to exhibit its virulence and to survive in periodontal pockets. The development of Rgp inhibitors thus provides new therapeutic approaches to periodontal diseases. In this study, we first isolated and purified a novel and potent inhibitor of Rgp from the culture supernatant of Streptomyces species strain FA-70, now designated as FA-70C1. This compound was found to be an antipain analog composed of phenylalanyl-ureido-citrullinyl-valinyl-cycloarginal (C27H43N9O7). The Ki value was calculated to be 4.5x10(-9) M when benzyloxycarbonyl-phenylalanyl-arginine-4 methly-coumaryl-7-amide was used as a substrate. This compound also inhibited cathepsins B, L, and H, though their Ki values were much higher than that of Rgp. FA-70C1 had little or no inhibitory activity on Lys-gingipain, another cysteine proteinase of P. gingivalis. The Rgp-induced degradation of various human proteins was completely blocked by this inhibitor. Disruption of both the bactericidal activity of polymorphonuclear leukocytes and the viability of human fibroblasts and umbilical vein endothelial cells induced by the culture supernatant of P. gingivalis was suppressed by the inhibitor in a dose-dependent manner. The enhancement of vascular permeability induced by in vivo administration of the culture supernatant of P. gingivalis was strongly inhibited by the inhibitor. Furthermore, the growth of P. gingivalis was suppressed by FA 70C1 in a dose-dependent manner. These results strongly suggest that FA-70C1 is a useful tool to prevent the virulence of P. gingivalis. PMID- 12887059 TI - Procongopain from Trypanosoma congolense is processed at basic pH: an unusual feature among cathepsin L-like cysteine proteases. AB - Congopain, the major cysteine protease from Trypanosoma congolense, is synthesized as an inactive zymogen, and further converted into its active form after removal of the proregion, most probably via an autocatalytic mechanism. Processing of recombinant procongopain occurs via an apparent one-step or a multistep mechanism depending on the ionic strength. The auto-activation is pH dependent, with an optimum at pH 4.0, and no activation observed at pH 6.0. After addition of dextran sulfate (10 microg/ml), an approx. 20-fold increase of processing (expressed as enzymatic activity) is observed. Furthermore, in the presence of dextran sulfate, procongopain can be processed at pH 8.0, an unusual feature among papain-like enzymes. Detection of procongopain and trypanosomal enzymatic activity in the plasma of T. congolense-infected cattle, together with the capacity of procongopain to be activated at weakly basic pH, suggest that procongopain may be extracellularly processed in the presence of blood vessel glycosaminoglycans, supporting the hypothesis that congopain acts as a pathogenic factor in host-parasite relationships. PMID- 12887060 TI - Attenuated kinin release from human neutrophil elastase-pretreated kininogens by tissue and plasma kallikreins. AB - Components of kinin-forming systems operating at inflammatory sites are likely to interact with elastase that is released by recruited neutrophils and may, at least temporarily, constitute the major proteolytic activity present at these sites. The aim of this work was to determine the effect of kininogen degradation by human neutrophil elastase (HNE) on kinin generation by tissue and plasma kallikreins. We show that the digestion of both low molecular mass (LK) and high molecular mass (HK) forms of human kininogen by HNE renders them essentially unsusceptible to processing by human urinary kallikrein (tissue-type) and also significantly quenches the kinin release from HK by plasma kallikrein. Studies with synthetic model heptadecapeptide substrates, ISLMKRPPGFSPFRSSR and SLMKRPPGFSPFRSSRI, confirmed the inability of tissue kallikrein to process peptides at either termini of the internal kinin sequence, while plasma kallikrein was shown to release the kinin C-terminus relatively easily. The HNE generated fragments of kininogens were separated by HPLC and the fractions containing internal kinin sequences were identified by a kinin-specific immunoenzymatic test after trypsin digestion. These fractions were analyzed by electrospray-ionization mass spectrometry. In this way, multiple peptides containing the kinin sequence flanked by only a few amino acid residues at each terminus were identified in elastase digests of both LK and HK. These results suggest that elastase may be involved in quenching the kinin-release cascade at the late stages of the inflammatory reaction. PMID- 12887061 TI - Effect of plant Kunitz inhibitors from Bauhinia bauhinioides and Bauhinia rufa on pulmonary edema caused by activated neutrophils. AB - Mediators released from polymorphonuclear neutrophils, in particular elastase, are known to induce acute edematous lung injury. In this study we show that the pulmonary edema in isolated perfused rabbit lungs caused by activated neutrophils via release of elastase is significantly decreased by the Kunitz-type Inhibitor BbCI (10(-5) M) from Bauhinia bauhinoides to the same degree as by eglin C (10( 5) M) from Hirudo medicinalis, which was used as a reference. The highly homologous proteinase inhibitor BrPI (10(-5) M) from Bauhinia rufa, however, did not reduce edema formation. The major difference between these inhibitors is the much higher Ki value of BrPI (Ki = 38 nM) for elastase compared to BbCI (Ki = 5.3 nM) and eglin C (Ki = 0.2 nM), respectively. Elastase liberation from activated PMNs was not influenced by the inhibitors. Our results indicate that BbCI can be a useful tool to study the role of neutrophil elastase in pathophysiological processes. PMID- 12887062 TI - Revisiting ubiquity and tissue specificity of human calpains. AB - The abundance of mRNAs transcribed from human genes of the calpain superfamily was studied in 72 human tissues and cell types by the Human Multiple Tissue Expression (MTE) Array technique. The analysis included the large subunits of mu- and m-calpains, the small subunits, calpastatin and calpain 3 (p94). Besides specific data on transcriptional activity, two major conclusions emerged: (i) 'ubiquitous' calpains are not expressed in all cell types, and (ii) a 'tissue specific' calpain may be expressed in many cell types apart from the one in which it is particularly abundant. Therefore, the categoric classification of 'ubiquitous' vs. 'tissue-specific' calpains is a simplification. PMID- 12887063 TI - The calpastatin-derived calpain inhibitor CP1B reduces mRNA expression of matrix metalloproteinase-2 and -9 and invasion by leukemic THP-1 cells. AB - The ubiquitous proteases mu- and m-calpain are Ca(2+)-dependent cysteine endopeptidases. Besides involvement in a variety of physio(patho)logical processes, recent studies suggest a pivotal role of calpains in differentiation of hematopoietic cells and tumor cell invasion. However, the precise actions of calpains and their endogenous inhibitor, calpastatin, in these processes are only partially understood. Here we have studied the role of the calpain/calpastatin system in the invasion of leukemic cells under basal and differentiation stimulating conditions. To further differentiate the human leukaemic cell line THP-1 (monocytic), the cells were treated for 24 hours with the differentiation stimulating reagents phorbol 12-myristate 13-acetate (PMA) and dimethyl sulfoxide (DMSO). Macrophage- and granulocyte-like differentiation was confirmed by induction of vimentin expression as well as by microscopic and fluorescence assisted cytometric analysis. Extracellular matrix (ECM) invasion of both the basal and differentiation-stimulated cells in a Matrigel assay was inhibited by pre-incubation of the cells with the specific calpain inhibitor CP1B for 24 hours. Inhibition of invasiveness correlated with decreased mRNA expression and secretion of the matrix metalloproteinases MMP-2 and MMP-9. In contrast, addition of CP1B only during the invasion process did neither influence transmigration nor MMP release. This is the first report showing that the calpain/calpastatin system mediates MMP-mRNA expression of the leukemic THP-1 cells and as a consequence their invasiveness. PMID- 12887064 TI - Heat stress-dependent DNA binding of Arabidopsis heat shock transcription factor HSF1 to heat shock gene promoters in Arabidopsis suspension culture cells in vivo. AB - Using UV laser cross-linking and immunoprecipitation we measured the in vivo binding of Arabidopsis heat shock transcription factor HSF1 to the promoters of target genes, Hsp18.2 and Hsp70. The amplification of promoter sequences, co precipitated with HSF1-specific antibodies, indicated that HSF1 is not bound in the absence of heat stress. Binding to promoter sequences of target genes is rapidly induced by heat stress, continues throughout the heat treatment, and declines during subsequent recovery at room temperature. The molecular mechanisms underlying the differences between Hsp18.2 and Hsp70 in the kinetics of HSF1/promoter binding and corresponding mRNA expression profiles are discussed. PMID- 12887065 TI - T47-D cells and type V collagen: a model for the study of apoptotic gene expression by breast cancer cells. AB - We have previously reported that type V collagen is a poorly adhesive, anti proliferative and motility-inhibitory substrate for the 8701-BC breast cancer cell line, which also triggers DNA fragmentation and impairs survival of the same cell line. In the present work we have extended to other breast cancer cell lines (T47-D, MDA-MB231, Hs578T) our investigation of type V collagen influence on the DNA status and cell survival, also examining whether adhesion and growth of cells on this collagen substrate could exert some effect on the expression level of selected apoptosis-related genes. We report here that, among the cell lines tested, only T47-D is responsive to the death-promoting influence of type V collagen. In addition, the latter induces changes in gene expression by up regulating p53, Waf-1, Cas, Dap kinase and caspases 1, -5 and -14 and down regulating Bcl-2. Our data validate the T47-D line as a suitable in vitro model for further and more detailed studies on the molecular mechanisms of the death response induced by type V collagen on mammary tumor cells. PMID- 12887066 TI - Cardiac catheterisation in infants weighing less than 2500 grams. AB - OBJECTIVES: To investigate the indications for, and outcome of, cardiac catheterisation in infants weighing less than 2500 g at a single institution over an 8-year period. PATIENTS AND METHODS: We assessed all infants who were less than 2500 g at the time of cardiac catheterisation at Texas Children's Hospital from January 1993 to January 2001. Comparisons of morbidity and mortality were drawn with an equivalent number of infants of similar age weighing greater than 2500 g seen over the same period of time. RESULTS: We performed interventional procedures in 22, and diagnostic catheterisations in 12 infants weighing less than 2500 g. Interventions included pulmonary valvoplasty in six patients, balloon angioplasty of critical coarctation in one, aortic valvoplasty in two, septostomy in ten, and coil occlusion of an arteriovenous malformation, redirection of a subclavian venous line, and coil occlusion of a patent arterial duct in one patient each. The median age at catheterisation was 5 days for children less than 2500 g, and 10 days for those above 2500 g. The median weights were 2.3 kg and 3.3 kg, and the median gestational ages were 35 weeks and 38 weeks, for the two respective groups. Of those weighing less than 2500 g, two died (6%), with no deaths occurring in those weighing more than 2500 g. In 3 patients weighing less than 2500 g (9%), there was vascular compromise, one child with bilateral femoral venous obstruction requiring fasciotomy compared, to one in the group weighing greater than 2500 g (2%). CONCLUSION: There is a significantly increased risk of mortality and vascular compromise in infants weighing less than 2500 g. Interventional catheterisation in these infants may be lifesaving, but given the aforementioned risks, diagnostic catheterisation should be deferred if possible in favor of noninvasive modalities. PMID- 12887067 TI - The ratio of flow in the superior and inferior caval veins after construction of a bidirectional cavopulmonary anastomosis in children. AB - In patients who have undergone a superior cavopulmonary anastomosis, the superior caval venous flow provides the only, or the most important, pulmonary blood supply, while the inferior caval venous blood is not oxygenated, being mixed with the pulmonary venous blood before entering the systemic circulation. In healthy children, the contribution of superior caval venous flow to total cardiac output has been shown to decrease during growth. Patients who have undergone a superior cavopulmonary anastomosis, however, often have a higher oxygen saturation than predicted by the age-matched ratio of superior to inferior caval venous flows. This study was designed, therefore, to assess the ratio of flows in the superior and inferior caval veins subsequent to a superior cavopulmonary anastomosis. We carried out 18 magnetic resonance imaging studies with velocity-mapping and heart catheterisations so as to assess the contribution of superior caval venous flow to total cardiac output. Patients were divided into 3 groups according to their age. There were five aged from 8 to 24 months, eight aged from 24 to 48 months, and five older than 48 months. No significant difference could be found in the ratios of superior-to-inferior caval venous flow, nor of superior caval venous-to systemic flow, between the 3 groups. The ratio of venous flows was 0.89 +/- 0.34 in those aged from 8 to 24 months, 1.09 +/- 0.42 in those from 24 to 48 months, and 1.25 +/- 0.27 in the older patients (F analysis of variance 1.06, p 0.37). The ratio of superior caval venous-to-systemic flow was 0.46 +/- 0.08 in the youngest patients, 0.50 +/- 0.09 in those aged from 24 to 48 months, and 0.55 +/- 0.05 in the older patients (F analysis of variance 0.76, p 0.49). These findings suggest that the hemodynamics of a cavopulmonary anastomosis may affect the normal decrease of superior caval venous flow with age. This could be related to a redistribution of flow, with a proportionally higher flow to the head and upper body after construction of a superior cavopulmonary anastomosis. Since increasing cyanosis and progressive exercise intolerance are the main indications for creation of a total cavopulmonary connection, these findings should be taken into account when determining the timing for completion of the Fontan circulation. PMID- 12887068 TI - Large pericardial effusions of inflammatory origin in childhood. AB - OBJECTIVES: Our aim was to review the clinical records from children with large pericardial effusions of inflammatory origin presenting to a tertiary referral centre over the last 21 years, with emphasis on their clinical presentation, management and outcome. BACKGROUND: The common identifiable causes of pericardial effusion in children include prior cardiac surgery, bacterial pericarditis, malignancy, and connective tissue disorders. In a significant number of children, however, despite extensive investigation, it is not possible to identify a clear aetiology. A viral cause is often considered, though rarely confirmed. The clinical course of such large idiopathic pericardial effusions in children has not been extensively reported. METHODS AND RESULTS: We reviewed retrospectively the records of all patients seen between 1981 and 2001 with large pericardial effusions of inflammatory origin requiring drainage, excluding the effusions related to cardiac surgery or malignancy. We found 31 patients fulfilling our criterions for study. They could be divided into three groups, with 15 patients having no specific identifiable aetiology despite extensive investigation, 12 patients having evidence of bacterial pericarditis, and four with a probable immunologic disorder. Fever was present in only eight patients (53%) in the idiopathic group. All patients in the other groups had fever. Except for fever and the resultant tachycardia, it was not possible to distinguish on clinical grounds, nor on the presence or otherwise of cardiac tamponade, between those with idiopathic aetiology and those with bacterial infection. Of the patients with presumed bacterial pericarditis, five (42%) had both positive blood and pericardial fluid cultures, three (25%) had positive blood cultures, while a further three patients (25%) had only positive pericardial fluid cultures. All patients required drainage of the pericardial effusion, either under echocardiographic guidance or surgically. None of the patients died. The hospital stay was significantly shorter for those with idiopathic as opposed to bacterial pericarditis. Of those with an idiopathic aetiology, six required readmission due to recurrence of the pericardial effusion, with four patients requiring further surgical drainage. No patients required readmission with a bacterial or immunologic aetiology. No patient developed constrictive pericarditis after a median follow-up of 22 months. CONCLUSION: Patients with large idiopathic pericardial effusion had relatively few constitutional symptoms as compared with their gross echocardiographic findings. Those with bacterial pericarditis had more urgent need for treatment. Patients with pericardial effusion of inflammatory origin, when treated appropriately, had an excellent outcome with no mortality or development of constrictive pericarditis. PMID- 12887069 TI - The influence of measured versus assumed uptake of oxygen in assessing pulmonary vascular resistance in patients with a bidirectional Glenn anastomosis. AB - AIMS: To determine the accuracy of data relating to pulmonary vascular resistance data in patients with a bidirectional Glenn anastomosis as calculated using predicted versus measured uptake of oxygen. METHODS: We studied retrospectively the data from 33 patients with a bidirectional Glenn anastomosis who underwent cardiac catheterisation prior to surgery to complete the Fontan circulation. Their weight ranged from 5.4 to 51.7 kg, and they were aged up to 12 years. Uptake of oxygen was measured using the Deltatrac II metabolic monitor. From the calculated indexed pulmonary vascular resistance, cases were stratified according to the risk of failure of the subsequent Fontan circulation. The six patients with a resistance of greater than 4 Um2 were deemed at high risk, the six with a resistance from 3 to 4 Um2 at moderate risk, and the 21 patients with a resistance less than 3 Um2 at low risk. Uptake of oxygen was also estimated from the predictive formulas of Lindahl, Lundell et al. and LaFarge and Miettinen. The indexed resistance was similarly calculated using these formulas and a comparable stratification of risk made from this data. RESULTS: The predicted values for uptake of oxygen were consistently higher than those measured, leading to an underestimation of indexed resistance, with mean difference between -0.62 and 1.57 Um2. This difference resulted in misclassification of between five and nine of the 12 patients considered at moderate or high risk as being at low-risk. No other haemodynamic data could reliably separate the subjects deemed at low-risk from those considered to be at high-risk. A transpulmonary gradient of greater than 7 mm of mercury was found to be 100 percent specific for elevated indexed resistance, but only 60 percent sensitive. CONCLUSIONS: In patients with bidirectional Glenn anastomoses, all formulas based on predictive uptake of oxygen lead to underestimation of the true indexed pulmonary vascular resistance, to an extent that could significantly influence clinical decision-making. The transpulmonary gradient is not a reliable surrogate for indexed pulmonary vascular resistance. PMID- 12887070 TI - Normal patterns of flow in the superior caval, hepatic and pulmonary veins as measured using Doppler echocardiography during childhood. AB - To date, no reference values have been provided for right and left atrial filling in normal children. The aim of our study, therefore, was to characterize measurements of superior caval, hepatic, and pulmonary venous flow using Doppler echocardiography in a large group of normal children to reflect the effects of age, body mass index, sex, heart rate and respiration. Doppler echocardiographic examinations of the superior caval, hepatic and pulmonary veins were performed during inspiration and expiration in 72 healthy children with a mean age of 6.73 +/- 5.10 years. The subjects were segregated into four age groups, namely infants < 2 years, preschool children between the ages of 2 and 7 years, children of school age between 7 and 11 years, and adolescents older than 11 years. Age has significant effect on the systolic and reverse atrial flows within the superior caval vein (p < 0.05). No change in the Doppler velocities was observed related to body mass index or sex. All peak systolic velocities decreased significantly during expiration (p < 0.05). This decrease was most prominent in the hepatic vein (26%), but less remarkable in the superior caval vein (5.7%) and the pulmonary veins (3.9%). During expiration, the peak diastolic flow in the superior caval and the hepatic veins decreased, while the reverse atrial flow in the hepatic vein increased (p < 0.05). Pulmonary venous velocities were similar in all age groups (p > 0.05). Except for the systolic pulmonary venous velocities, these parameters were not influenced by respiration (p > 0.05). The diastolic time, the interval between reverse atrial flow and ventricular systole reflected by the R wave on the electrocardiogram, and the interval between ventricular systole and diastolic flow, were negatively correlated with heart rate (p < 0.05; r = -0.35, -0.85, and -0.8 respectively), and positively correlated with age (p < 0.05; r = 0.3, 0.8, and 0.7 respectively). They were not influenced by respiration. Our study provides data of the patterns and the normal ranges of velocities of superior caval, hepatic, and pulmonary venous flow in a series of normal children. The results can now be used for comparison with the patterns found in the setting of disease. PMID- 12887071 TI - A model for left juxtaposition of the atrial appendages in the chick. AB - The morphogenesis of most types of human congenital cardiac malformations is still obscure. The reasons for this are, first, the paucity of data from human embryos and fetuses and, second, the paucity of appropriate animal models. During the past few years, we have tested several chemicals for their teratogenic potential, hoping to find, particularly in the chick, substances that could be used for the development of models for specific cardiac malformations. We have now discovered that suramin, an antitrypanosomal drug, can induce a special type of congenital cardiac defect in which the two atrial appendages are positioned to the left of the great arteries. This situation resembles the situation found in humans and classified as left juxtaposition of the atrial appendages. In the present study, we have analysed the pathomorphological features of a series of our chicken hearts to assess precisely how accurately these cardiac malformations in the chick correspond to the situation seen in the human. We found that the cases observed in the chick did, indeed, have many features in common with the human cases described in the literature. This suggests that we have developed an animal model for human left juxtaposition. Our model could be used for two kinds of embryological studies: first, documentation of the morphogenesis of left juxtaposition; and second, studies on the mechanisms driving the normal positional changes between the atriums and outflow tract of the embryonic heart during the late phase of cardiac looping. The fact that left juxtaposition of the atrial appendages can be induced by suramin might help to elucidate the molecular mechanisms underlying this type of congenital cardiac malformation. Furthermore, the fact that suramin is used for the chemotherapy of frequent tropical diseases, such as African trypanosomiasis and onchocerciasis, poses the question as to whether this drug might play a role in the aetiology of left juxtaposition in some human populations. PMID- 12887072 TI - Kinetics of procalcitonin, interleukin 6 and C-reactive protein after cardiopulmonary-bypass in children. AB - Cardiopulmonary bypass induces a generalized inflammatory response, with fever and leukocytes, which is difficult to differentiate from an infection. Recently, procalcitonin has been proposed as an early and specific marker of bacterial infection. The influence of cardiopulmonary bypass on production of procalcitonin, therefore, must be assessed before considering this molecule as a valuable marker of infection after cardiac surgery in children. With this in mind, we measured levels of procalcitonin, interleukin 6, and C-reactive protein before and 6h, 1, 3 and 5 days after cardiopulmonary bypass, in 25 children undergoing cardiac surgery. Cardiopulmonary-bypass induced a transient increase in procalcitonin, with a peak at 24 h, with a median of 1.13 microg/l, a 25th and 75th interquartile of 0.68-2.25, and a p value of less than 0.001. The value had returned to normal in the majority of the children by the third day after surgery. Peak values correlated with the duration of cardiopulmonary-bypass, with a r-value of 0.58 and a p value of 0.003; cross-clamp time, with a r-value of 0.62 and a p value of 0.001; days of mechanical ventilation, with a r-value of 0.62 and a p value of 0.001; and days of stay in intensive care, with a r-value of 0.68, and a p value of 0.0003. The value returned to normal after 3 days in 83% of the patients. Levels of interleukin 6 and C-reactive protein also increased significantly after surgery, and remained elevated for up to 5 days. Thus, in contrast to other markers, levels of procalcitonin in the serum are only slightly and transiently influenced by cardiopulmonary bypass, and may prove to be useful in the early recognition of an infection subsequent to cardiopulmonary bypass. PMID- 12887073 TI - Echocardiographic presentations of endocarditis, and risk factors for rupture of a sinus of Valsalva in childhood. AB - BACKGROUND: In recent years, the diagnosis of infective endocarditis has been enhanced by the use of echocardiography. We sought, therefore, to review its effect on the management of endocarditis in children. METHODS: We reviewed all the patients presenting to our institution for evaluation for infective endocarditis from May 1994 to January 2002. The patients were stratified according to whether or not they had congenitally malformed hearts. RESULTS: Of the 90 referred patients identified, 46 (51%) had positive ultrasonic findings. Of these, we excluded 26 patients because of the presence of indwelling lines. The remaining 20 patients with features of endocarditis had a median age of 6.5 years, and a range from 0.14 to 8.5 years. There were 4 patients with normal hearts, and 16 with congenital cardiac malformations. We identified rupture of a sinus of Valsalva in four patients, with rupture into the left ventricle in two, and into the right ventricle and right atrium in one each. The mitral valve was involved in six patients, the aortic valve in another six, including all four with rupture of the sinus of Valsalva, both mitral and aortic valves in three, the pulmonary trunk in three patients, and the tricuspid valve and a Blalock Taussig shunt in one patient each. Organisms isolated included Streptococcus mitis in 4 patients, Streptococcus pneumoniae in 2 patients, Streptococcus sanguis in 1, Staphylococcus aureus in 3, Staphylococcus epidermidis in 1, and Enteroccocus in 2. Cultures proved negative in 7 patients. Surgical intervention was needed in 12 patients, and one died (5%). Only the left-sided chambers were involved in those with normal hearts. Both patients infected with Streptococcus pneumoniae had rupture of a sinus of Valsalva. CONCLUSION: Involvement of the left-sided chambers is more likely in structurally normal hearts, and in cases with rupture of a sinus of Valsalva, in which case infection with Streptococcus pneumonia should be suspected. PMID- 12887074 TI - Rapid disappearance of a huge cardiac rhabdomyoma in an infant. PMID- 12887075 TI - Mannheimer Lecture. The quintessence of the making of the heart. AB - In my Mannheimer lecture, designed to meet the needs of a mainly clinical audience, I present aspects of cardiac development that link basic science to clinically relevant problems. During development of the cardiac tube, and its subsequent changes as a dextrally looped structure, which is still connected to the dorsal body wall by a venous and an arterial pole, there are basic requirements. These consist of the development of myocardium, endocardium and the interposed cardiac jelly from the cardiogenic plates. In this primitive heart tube, septation and valvar formation then take place to convert it into a four chambered heart. I demonstrate that the refining of the above events cannot take place without the addition of extracardiac populations of cells. These are presented as the "quintessence of heart development", and consist of cells derived from the neural crest, along with epicardially derived cells. Without these contributions, the embryos uniformly die of cardiac insufficiency. Important contributions are made by the cells derived from the neural crest to septation and the formation of the arterial valves, and possibly in differentiation of the central conduction system. The epicardially derived cells are essential for formation of the interstitial fibroblasts and the myocardium, as well as the coronary vascular system. I conclude by discussing specific malformations of the heart that might be linked to these extracardiac contributions. PMID- 12887076 TI - The vestibular defect: an interatrial communication due to a deficiency in the atrial septal component derived from the vestibular spine. PMID- 12887077 TI - Replacement of the aortic valve after the arterial switch operation. AB - An 11-month-old infant had undergone a primary arterial switch operation, including the Lecompte maneuver, for correction of discordant ventriculo-arterial connections and closure of an accompanying ventricular septal defect. At discharge, there were no signs of aortic valvar incompetence. Regurgitation across the aortic valve was detected first at the age of 2 years, and then increased progressively, as documented by serial echocardiographic studies. There had been no history of bacterial endocarditis. At the age of 10 years, echocardiography revealed severe aortic valvar incompetence. At operation, the aortic valve had three leaflets, all of which were short, with very restricted movement. Absence of sufficient leaflet tissue precluded a durable valvar reconstruction, so the aortic valve was replaced with a 21 mm mechanical prosthesis. Histological examination of the removed leaflets revealed nodular swelling due to mucous changes of the matrix, as well as fibrous alteration and formation of scar tissue, including areas of fibroblastic and capillary proliferation. There were no signs of calcification or acute inflammatory changes. Improvement of left ventricular function was observed both early postoperatively and later on. Our observation shows that aortic valvar incompetence after an arterial switch operation can be caused by degenerative changes of the neo-aortic leaflets, which prevent plastic reconstruction of the valve, necessitating valvar replacement. PMID- 12887078 TI - Prenatal diagnosis of tetralogy of Fallot associated with a fistula from the left coronary artery to the left atrium. AB - In a fetus at 20 weeks gestation, we found a large fistula from the left coronary artery to the left atrium in association with tetralogy of Fallot. Postnatally, the therapeutic issues were complex because of prematurity, low birth weight, decreased flow of blood to the lungs, and volume overload of the left ventricle because of the huge fistula. At three months, the baby underwent repair of tetralogy of Fallot, with surgical ligation of the fistula, but the baby died postoperatively. PMID- 12887079 TI - Histopathological follow-up after bailout stenting for early postoperative stenosis of a central aorto-pulmonary shunt. AB - We treated a neonate with pulmonary atresia and a ventricular septal defect complicated by straddling of the atrioventricular valves by constructing a central aorto-pulmonary shunt. The postoperative course was complicated by obstruction of the shunt, which was treated by implantation of a coronary stent. Six months after the stenting, a Glenn anastomosis was created and the stented shunt removed. Analysis showed that the shunt was completely covered by a vascularized neointima. The stent had not produced injury to the shunt, with struts of the stent covered nicely by neoendothelium, with sparse inflammation surrounding the artificial implants. PMID- 12887080 TI - Transcatheter closure of a multiperforated atrial septal defect extending from the oval fossa to the mouth of the inferior caval vein. AB - We report a novel technique using an Amplatzer atrial septal occluder to close a defect located in the inferior-posterior portion of the interatrial septum that extended into the mouth of the inferior caval vein. Because of the close relation of the defect to the inferior caval vein, the right atrial disc was opened into the inferior caval vein and pushed toward the right atrium by use of the delivery cable. There was no residual shunting immediately and 3 months after the intervention. We conclude that even defects located infero-posteriorly within the oval fossa may be successfully closed by transcatheter techniques using the Amplatzer device. PMID- 12887081 TI - Unusual interventional management in an adult with tetralogy of Fallot. AB - A 53-year-old man with tetralogy of Fallot had been palliated with two classic Blalock-Taussig shunts. Cardiac catheterization in our center revealed acquired atresia of the pulmonary valve. We perforated and dilated the valve as a palliative procedure with a new system using radiofrequency energy. PMID- 12887082 TI - Methotrexate in recurrent postpericardiotomy syndrome. AB - The postpericardiotomy syndrome occurs in up to one-third of children undergoing cardiac surgery. Its treatment includes anti-inflammatory agents, diuresis, and drainage of effusions. Administration of steroids can have a dramatic effect, but is limited by adverse effects. Usually the syndrome lasts weeks only, and persistence beyond six months is exceptional. We describe a rare case of chronic postpericardiotomy syndrome, with recurrent pericardial effusions and steroid dependency, that was treated successfully with a low weekly dose of methotrexate. PMID- 12887083 TI - Ischemia due to peripartum cardiomyopathy threatening loss of a leg. AB - Ischemia of the leg in a peripartum female is an uncommon condition. Paradoxical arterial embolisation, and arterial dissection, are rarely encountered but recognized causes of this clinical condition. Peripartum cardiomyopathy is a rare life-threatening cardiac condition that can foster intracardiac thrombosis and produce peripheral vascular complications through embolization. We present here the case of a young, healthy, postpartum female who developed acute ischemia of the left leg, and asymptomatic arterial insufficiency of the right lower leg, as the presenting symptoms of peripartum cardiomyopathy, highlighting the fact that ischemia threatening the loss of a limb can be the initial manifestation of peripartum cardiomyopathy. PMID- 12887084 TI - Signaling pathways in sperm capacitation and acrosome reaction. AB - The binding to the egg's zona pellucida stimulates the spermatozoon to undergo acrosome reaction, a process which enables the sperm to penetrate the egg. Prior to this binding, the spermatozoa underago in the female reproductive tract a series of biochemical transformations, collectively called capacitation. The first event in capacitation is cholesterol efflux leading to the elevation of intracellular calcium and bicarbonate to activate adenylyl cyclase (AC) to produce cyclic-AMP, which activates protein kinase A (PKA) to indirectly phosphorylate certain proteins on tyrosine. During capacitation, there is also an increase in protein tyrosine phosphorylation dependent actin polymerization and in the membrane-bound phospholipase C (PLC). Sperm binding to zona-pellucida causes further activation of cAMP/PKA and protein kinase C (PKC), respectively. PKC opens a calcium channel in the plasma membrane. PKA together with inositol trisphosphate activate calcium channels in the outer acrosomal membrane, which leads to an increase in cytosolic calcium. The depletion of calcium in the acrosome will activate a store-operated calcium entry mechanism in the plasma membrane, leading to a higher increase in cytosolic calcium, resulting in F-actin dispersion which enable the outer acrosomal and the plasma membrane to come into contact and fuse completing the acrosomal reaction. PMID- 12887085 TI - Role of signaling pathways in regulating the capacitation of mammalian spermatozoa. AB - Mammalian testicular spermatozoa are immotile and incompetent for fertilization. They acquire motility during epididymal maturation and fertilizing ability during a second phase of maturation in the female reproductive tract, termed as capacitation. Capacitation was discovered independently by Austin and Cang in early 1950s and was defined as the obligate period of residency of spermatozoa in the female reproductive tract, which confers on the spermatozoa the ability to fertilize an oocyte. Over the years, the definition of capacitation has changed and it has been recognized as a complex phenomenon, which is correlated with changes associated with the spermatozoa in the female tract. These alterations in metabolism, intracellular ion concentration, membrane fluidity, intracellular pH, cAMP concentration and concentration of reactive oxygen species, ultimately make the spermatozoa fertilization-competent. The molecular basis of capacitation is poorly understood despite the fact that it is an important event preceding fertilization. This review presents our current understanding of the signaling events involved in the process of capacitation. PMID- 12887086 TI - Developmental changes in signalling transduction factors in maturing sperm during epididymal transit. AB - Testicular spermatozoa are incapable of fertilising eggs in vivo and the epididymal epithelium is responsible for creating a luminal environment conducive to the acquisition of fertilising capacity by the spermatozoa within it. It does this by modifying sperm membranes so that they can interact with stimuli from the female tract, and by supplying or removing low molecular weight compounds that modify sperm cytoplasmic composition. Both these extracellular and intracellular events associated with signal transduction and second messenger systems in maturing spermatozoa are reviewed here. PMID- 12887087 TI - Hyperactivation of mammalian sperm. AB - Mammalian sperm commonly show hyperactivated motility just before fertilization. The movement of hyperactivated sperm appears different in fluids of different viscosity and elasticity and in different species, but basically it involves an increase in flagellar bend amplitude and, usually, beat asymmetry. Hyperactivation may be critical to the success of fertilization, because it enhances the ability of sperm to detach from the wall of the oviduct, to move around in the labyrinthine lumen of the oviduct, to penetrate mucous substances and, finally, to penetrate the zona pellucida of the oocyte. Presumably, a signal or signals exist in the oviduct to initiate hyperactivation at the appropriate time; however, none have yet been identified with certainty. While the signal transduction cascade regulating hyperactivation remains to be completely described, it is clear that calcium ions interact with the axoneme of the flagellum to switch on hyperactivation. Although hyperactivation often occurs during the process of capacitation, divergent pathways regulate the two events. PMID- 12887088 TI - How do sperm swim? Molecular mechanisms underlying sperm motility. AB - Motility is a characteristic function of the male gamete, which allows spermatozoa to actively reach and penetrate the female gamete in organisms with internal and external fertilization. Sperm motility is acquired under the control of many extrinsic and intrinsic factors and is based on the specialized structure of the sperm flagellum. After a brief overview of how the sperm flagellum is organized and works to support cell motility, the present review focuses on the molecular mechanisms and factors involved in the development and maintenance of sperm motility. Data obtained both in organisms with external fertilization, such as fishes and sea urchin, and with internal fertilization, such as Mammals, are critically analyzed. In particular, a great attention has been put on the ionic mechanisms and on the involvement of protein kinases and phosphatases in regulation of sperm motility. A brief overview of the pharmacological and physiological molecules which have been studied for their possible application as therapeutic molecules for in vitro treatment of defects of sperm motility in asthenozoospermic human subjects, is presented. Moreover, we show some preliminary data obtained in our laboratory on the involvement of the phosphatydilinositol 3-kinase and the A kinase anchoring protein (AKAP3) in regulation of motility in human spermatozoa. The last section is dedicated to hyperactivation, a peculiar pattern of motility which is developed in association with capacitation occurring during sperm transit through the female genital tract and which can also be obtained in vitro by incubation in defined media. PMID- 12887089 TI - Pentoxifylline induced signalling events during capacitation of hamster spermatozoa: significance of protein tyrosine phosphorylation. AB - To fertilize the oocyte, mammalian spermatozoa must undergo capacitation and acrosome reaction. These events are believed to be associated with various biochemical changes primarily mediated by cAMP, Ca2+ and protein kinases. But the precise signaling mechanisms governing sperm function are not clear. To study this, we used pentoxifylline (PF), a sperm motility stimulant and a cAMP phosphodiesterase inhibitor, during capacitation and acrosome reaction of hamster spermatozoa. PF induced an early onset of sperm capacitation and its action involved modulation of sperm cell signaling molecules viz, cAMP, [Ca2+]i and protein kinases. The PF-induced capacitation was associated with an early and increased total protein phosphorylation coupled with changes in the levels of reactive oxygen species. Protein kinase (PK)-A inhibitor (H-89) completely inhibited phosphorylation of a 29 kDa protein while PK-C inhibitor (staurosporine) did not inhibit phosphorylation. Interestingly, PF induced protein tyrosine phosphorylation of a set of proteins (Mr 45-80 K) and a greater proportion of PF-treated spermatozoa exhibited protein tyrosine phosphorylation, compared to untreated controls (82 + 9% vs 34 +/- 10%; p < 0.001); tyrosine phosphorylated proteins were localized specifically to the mid-piece of the sperm. The profile of protein tyrosine phosphorylation was inhibitable by higher concentrations (> 0.5 mM) of tyrosine kinase inhibitor, tyrphostin A47. However, at lower (0.1-0.25 mM) concentrations, the compound interestingly induced early sperm capacitation and protein tyrosine phosphorylation, like PF. These results show that protein tyrosine phosphorylation in the mid-piece segment (mitochondrial sheath) appears to be an early and essential event during PF induced capacitation and a regulated level of tyrosine phosphorylation of sperm proteins is critical for capacitation of hamster spermatozoa. PMID- 12887090 TI - CAMP activates Rap1 in differentiating mouse male germ cells: a new signaling pathway mediated by the cAMP-activated exchange factor Epac? AB - Rap1, a Ras-like G-protein, is implicated in the signaling of various cellular processes as morphogenesis, differentiation, cell adhesion and spreading, and maintenance of T cell anergy and B cell activation. The effectors that mediate Rap1 signaling have not yet been definitely identified, with the exception of B Raf which, however, is restricted to neuronal tissues and a small subset of other cell types, including in particular male germ cells. We previously showed that in mouse spermatids Rap1 could interact with B-Raf giving rise to a signaling complex. Here we investigated about the possible molecules which "switch on" Rap1 finding that cAMP could in vivo activate endogenous Rap1. Spermatid-enriched cell cultures stimulated with 8-(4-chlorophenylthio)-cyclic AMP yielded higher levels of GTP-bound Rap1 than unstimulated cells. Since cAMP-induced Rap1 activation is actually retained to occur through Epac, we checked whether this recently discovered Rap1 exchange factor is expressed in male germ cells. Our findings indicate that Epac is present in spermatogenic cells and exhibits a preferential subplasmalemmal localization, although it shows also an intracellular location, more or less pronounced depending on the type of spermatogenic cell examined. Taken together, our data show that cAMP activates Rap1 in differentiating male germ cells which express the cAMP sensor Epac, thus suggesting that this activation might occur directly through Epac. PMID- 12887091 TI - Glycosylation of zona pellucida glycoprotein-3 is required for inducing acrosomal exocytosis in the bonnet monkey. AB - To investigate the role of polypeptide backbone vis-a-vis glycosylation of the putative primary sperm receptor, the bonnet monkey (Macaca radiata) zona pellucida glycoprotein-3 (bmZP3), excluding the N-terminal signal sequence and the C-terminal transmembrane-like domain, was expressed as polyhistidine fusion protein either in Escherichia coli orusing baculovirus expression system. The recombinant bmZP3 (r-bmZP3) was purified using nickel-nitrilotriacetic acid resin and subsequently refolded in the presence of oxidized and reduced glutathione. SDS-PAGE and Western blot analysis revealed approximately 43 and approximately 52 kDa bands corresponding to E. coli and baculovirus expressed r-bmZP3, respectively. The r-bmZP3 purified from both E. coli and baculovirus binds to the principal segment of the acrosomal cap of the capacitated bonnet monkey spermatozoa as evaluated by indirect immunofluoresence assay. In a competitive inhibition assay, the binding of biotinylated baculovirus expressed r-bmZP3 to capacitated spermatozoa was inhibited not only by cold baculovirus expressed r bmZP3 but also by E. coli expressed r-bmZP3 and vice-versa. Lectin binding studies revealed that the baculovirus r-bmZP3 has N-linked glycosylation with galactose and mannose residues. Capacitated spermatozoa, in the presence of baculovirus expressed r-bZP3, undergoes a significant increase (p < 0.01) in the acrosomal exocytosis as compared to control whereas E. coli expressed r-bmZP3 failed to have a significant effect. These results suggest that though the polypeptide backbone of ZP3 is sufficient for its binding to capacitated spermatozoa, yet glycosylation is required for induction of acrosomal exocytosis. PMID- 12887092 TI - Polo-like kinase-1 in porcine oocyte meiotic maturation, fertilization and early embryonic mitosis. AB - Polo-like kinases (Plks) are a family of serine/threonine protein kinases that regulate multiple stages of mitosis. Expression and distribution of polo-like kinase 1 (Plk1) were characterized during porcine oocyte maturation, fertilization and early embryo development in vitro, as well as after microtubule polymerization modulation. The quantity of Plk1 protein remained stable during meiotic maturation. Plk1 accumulated in the germinal vesicles (GV) in GV stage oocytes. After germinal vesicle breakdown (GVBD), Plk1 was localized to the spindle poles at metaphase I (MI) stage, and then translocated to the middle region of the spindle at anaphase-telophase I. Plk1 was also localized in MII spindle poles and on the spindle fibers and on the middle region of anaphase telophase II spindles. Plk1 was not found in the spindle region when colchicine was used to inhibit microtubule organization, while it accumulated as several dots in the cytoplasm after taxol treatment. After fertilization, Plk1 concentrated around the female and male pronuclei. During early embryo development, Plk1 was found to be in association with the mitotic spindle at metaphase, but distributed diffusely in the cytoplasm at interphase. Our results suggest that Plk1 is a pivotal regulator of microtubule organization and cytokinesis during porcine oocyte meiotic maturation, fertilization, and early embryo cleavage in pig oocytes. PMID- 12887093 TI - Studies on the participation of epididymal sperm protein DE/CRISP-1 in egg activation. AB - Protein DE (32 kDa) associates with sperm during epididymal maturation and participates in sperm-egg fusion through its binding to complementary sites on the egg surface. In the present work we investigated the participation of DE in two mechanisms probably involved in egg activation: the ability of DE to trigger activation by its interaction with the binding sites on the egg surface (receptor model) and its ability to regulate intracellular calcium channels (sperm factor model). The incubation of eggs with DE did not promote activation parameters such as calcium oscillations or meiosis resumption. Secondly, microinjection of DE into eggs was ineffective in either eliciting calcium release or modifying oscillations induced by an activating sperm extract. Together, these results argue against the participation of DE in egg activation, restricting the activity of this protein and its egg binding sites to the sperm-egg fusion process. PMID- 12887094 TI - Isolation and identification of a novel motility-inhibiting factor from goat cauda sperm plasma membrane. AB - A motility inhibiting factor (MIF) in sperm plasma membrane of mammalian spermatozoa (goat) has been demonstrated. This factor has been purified to apparent homogeneity by Sepharose-6B affinity chromatography and DEAE-cellulose ion-exchange chromatography. The molecular weight of the isolated factor has been estimated as 98 kDa by molecular sieving and analytical HPLC. SDS-polyacrylamide gel electrophoresis of MIF gave a single band of 100 kDa, indicating that the factor is a monomer. MIF is a thermo-stable factor and it inhibited the spermatozoa motility in a dose dependent manner. It is a glycoprotein as it binds with high affinity to Sepharose-6B and the affinity matrix-bound factor can be eluted with D-galactose. Data show that the motility inhibiting activity is lost completely when treated with beta-galactosidase indicating that its sugar side chain is essential for its activity. Addition of MIF antibody caused significant enhancement of forward motility of the caput and cauda-spermatoza. This antibody may thus be useful for solving some of the problems of human infertility due to low sperm motility. The motility inhibiting protein may also be useful as a vaginal contraceptive. PMID- 12887095 TI - Inhibition of in vitro capacitation of hamster spermatozoa by nitric oxide synthase inhibitors. AB - In an attempt to understand the role of nitric oxide(NO) in sperm capacitation, in the present study, hamster spermatozoa were used to evaluate the effects of NO on motility, viability, hyperactivation, capacitation and protein tyrosine and serine phosphorylation using specific inhibitors of nitric oxide synthase (NOS); namely L-NAME (N-nito-L-aginine methyl ester) and 7-Ni (7-nitroindazole). The results indicated that L-NAME inhibits sperm motility, hyperactivation and acrosome reaction where as 7-Ni inhibits only hyperactivation and acrosome reaction thus implying that NOS inhibitors exhibit subtle differences with respect to their effects on sperm functions. This study also provides evidence that NOS inhibitors inhibit sperm capacitation by their ability to modulate protein tyrosine phosphorylation. However, the inhibitors had no effect on the protein serine phosphorylation of hamster spermatozoa during capacitation. Thus, these results indicate that NO is required PMID- 12887096 TI - Vasoactive intestinal peptide and pituitary adenylate cyclase-activating polypeptide: players in innate and adaptive immunity. AB - Recent reports identified and described neural pathways, both hard-wiring and soluble mediators, that control and adjust the peripheral immune response. Immune organs are innervated by fibers rich in neurotransmitters and neuropeptides released in inflammatory conditions. Here we focus on the immunomodulatory role of two peptides, the vasoactive intestinal peptide (VIP) and the pituitary adenylate cyclase-activating polypeptide (PACAP). VIP/PACAP are present and released from both innervation and immune cells, particularly Th2 cells, and immune cells express receptors for VIP/PACAP. VIP/PACAP have a general anti inflammatory effect, both in innate and adaptive immunity. In innate immunity, VIP/PACAP inhibit the production of pro-inflammatory cytokines and chemokines from macrophages, microglia and dendritic cells. In addition, VIP/PACAP reduce the expression of costimulatory molecules (particularly CD80 and CD86) on the antigen-presenting cells, and therefore reduce stimulation of antigen-specific CD4+ T cells. In terms of adaptive immunity, VIP/PACAP promote Th2-type responses, and reduce the pro-inflammatory Th1-type responses. Several of the molecular mechanisms involved in the inhibition of cytokine and chemokine expression, and in the preferential development and/or survival of Th2 effects, are discussed. PMID- 12887097 TI - Modulation of regulatory T cell immunity by the neuropeptide alpha-melanocyte stimulating hormone. AB - Although many immunosuppressive factors have been identified in the eye, one of these factors, alpha-melanocyte stimulating hormone (alpha-MSH), both suppresses the activation of inflammatory activity by primed T cells and induces the activation of regulatory T cells (Treg cells). This neuropeptide alone at its ocular physiological concentration can account for most of the immunosuppressive activity of aqueous humor (the fluid filing the anterior chamber of the eye). Aqueous humor made devoid of alpha-MSH no longer suppresses IFN-gamma production by Th1 cells. It is alpha-MSH that mediates aqueous humor induction of regulatory T cells. What we have found is that alpha-MSH mediates the induction of C4+ CD25+ Treg cells, and that if the alpha-MSH Treg cells are specific to an autoantigen they can be used to suppress autoimmune disease. It is the objective of this review to demonstrate how we came to discover that alpha-MSH could have such an important role in the extreme regional immunity of the immune privileged eye and how this discovery could be applied to create or reestablish tolerance to prevent autoimmune disease. PMID- 12887098 TI - Aerosolized substance P protects against cigarette-induced lung damage and tumor development. AB - Both first-hand and second-hand [i.e. side-stream cigarette smoke (SSCS)] exposure to cigarette smoke is known to damage the lungs, alter the immune system, and predispose individuals to the development of emphysema and lung cancer. Previous work from these laboratories has demonstrated that administration of aerosolized substance P (SP) was capable of protecting the pulmonary and immune systems from damage due to environmental toxicants (e.g. hydrocarbon exposures). In the present study, the effects of SP on SSCS exposure were examined. Treatment of mice exposed to SSCS with aerosolized SP prevented pathological cellular and functional changes in the lung as reflected by prevention of damage to airway basement membranes/endothelial cells and preservation of normal airway dynamic compliance. Further, SP treatment reduced and/or prevented the occurrence of micronuclei formation in cells isolated from mice exposed in vivo to SSCS (an indicator of DNA/genetic damage). Finally, in an experimental in vivo lung cancer model, SP therapy significantly reduced the numbers of lung tumors, increased animal survival, and activated pulmonary immune defense mechanisms. Thus, aerosolized SP therapy appears to be capable of inhibiting preventing and/or reversing the cellular and genetic precursors of emphysema and malignancy that often result from exposure to cigarette smoke. PMID- 12887099 TI - Regulation of type 1 diabetes by a self-MHC class II peptide: role of transforming growth factor beta (TGF-beta). AB - The present study was undertaken to analyze the regulatory T cells generated in response to class I derived self-I-A beta(g7) (54-76) peptide. It was observed T cells from young unprimed type 1 diabetes (T1D) prone NOD mice did not respond to self-I-A beta(g7) (54-76) peptide although T cells from primed young NOD mice showed a strong response. T cells from young unprimed BALB/c mice responded to self-I-A beta(d) (62-78) peptide. However, a breakdown of tolerance to these peptides was observed with age in both the strains. Culture supernatant from I-A beta(g7) (54-76) peptide-primed cells secreted large amounts of TGF-beta and inhibited T cell responses in allogeneic-MLR. Further, I-A beta(g7) (54-76) peptide specific T cell lines from young (I-A.Y) and diabetic (I-A.D) NOD mice were established. I-A.Y secreted IL-4, TGF-beta and IL-10 while I-A.D T cell line secreted IL-10 and IFN-gamma. We found that I-A.D T cell line induced diabetes when transferred in NOD/SCID mice but I-A.Y T cell line did not induce disease. These results show that immunization of NOD mice with I-A beta(g7) (54-76) peptide at a younger age induces a regulatory T cell response suggesting that correcting the defects in immunoregulatory mechanisms using self-MHC peptides may be one of the approaches to prevent autoimmune diseases like T1D. PMID- 12887100 TI - Immune control of coxsackievirus-induced cardiopathology. AB - Retrovirally induced acquired immunodeficiency in humans and mice induces immune dysregulation as well as increased oxidative stress as the disease progresses. Both immunodeficiency and oxidative stress make the host susceptible to the development of heat disease either by physiological changes or by the direct influence of cardiovirulance. Antioxidant supplementation has been shown to influence the onset as well as the degree of cardiopathology due to primary infections or co-infections. An alternative treatment is the use of peptide immunomodulatory therapy to enhance cytokine production, immune cell function and resistance to opportunistic infections. This review compiles our in vivo and in vitro studies on the effects of antioxidant supplementation and peptide therapy on the immune control of coxsackievirus induced cardiopathology in AIDS. PMID- 12887101 TI - Immunoregulation by Vbeta specific antibodies in myasthenia gravis: mining physiological T cell homeostasis for TCR specific therapy. AB - Besides regulatory T cells, also comprising T cell receptor (CR)-specific T cells, it is increasingly evident that natural autoantibodies, among which anti TCR antibodies represent additional immunomodulators in the immune system. We took advantage of myasthenia gravis (MG), a well-characterized antibody-mediated autoimmune disease, to demonstrate that without prior vaccination against TCR determinants, patients with MG present increased circulating anti-TCR antibodies directed to the dominant TCR used by pathogenic T cells. These findings, pointing to a regulatory protective role of anti-TCR antibodies, are discussed in the context of the mechanisms of action and the physiological role of anti-TCR antibodies in T cell homeostasis, and of the puzzling world of regulatory T cells. Natural anti-TCR antibodies are found in the serum of all individuals, with prevalence in physiological and pathological situations such as ageing, pregnancy, allograft transplantation, retroviral infection, and autoimmune diseases, including MG. The common link is the mounting of immune responses against alloantigens, pathogens or autoantigens, conferring on anti-TCR antibodies a broader role in controlling responses to any antigen (self or non self) and more generally in T cell homeostasis. This homeostasis mechanism may well be exploited in therapeutic strategies based on TCR peptide vaccination in autoimmune diseases. PMID- 12887102 TI - Natural autoantibodies to TCR public idiotopes: potential roles in immunomodulation. AB - Autoantibodies directed against variable domain epitopes of the alpha/beta T cell receptor (TCR) occur in sera of man, mouse and other vertebrates. Here, we focus upon autoantibodies expressed in human rheumatoid arthritis (RA) and systemic erythematosus (SLE) with parallel studies involving collagen induced arthritis (CIA) in mice transgenic for human HLA-DR conferring resistance or susceptibility to autoimmune disease. We report specificity characterization of polyclonal and monoclonal IgM and IgG autoantibodies from SLE and for IgM monoclonal autoantibodies of RA patients. The data suggests that autoantibodies directed against "public" idiotopes present in the first complementarity determining region (CDR1) and the third framework (FR3) of the Vbeta gene products are generated in response to over-production of autodestructive T cells bearing particular Vbeta gene products and function to modulate (downregulate) the expression of these T cells. Since antibodies of these specificities are present in polyclonal IgG immunoglobulin (IVIG) preparations used for therapeutic purposes, the immunomodulatory effects of antibodies directed against TCR variable domains may account, at least in part, for the efficacy of IVIG preparations in therapy of autoimmune diseases and in the prevention of graft versus host reactions. PMID- 12887103 TI - Antibody like peptidomimetics as large scale immunodetection probes. AB - Antibodies are often used to study the molecular basis of physiologic processes. Despite the widespread applications of monoclonal antibodies (mAb) from basic science to successful therapeutics in clinical settings their use is limited. Production of mAb is often cumbersome and creating diverse and therapeutic amounts of useful mAb is difficult. We have developed a methodology to reduce an antibody into much smaller peptidomimetics and have engineered the mimetics for increased serum half life and affinity. The novel species are termed "antibody like binding peptidomimetics" (ABiP). We developed the Anti-Her2/neu peptidomimetic (AHNP) which is a mimic of Herceptin, a mAb used for advanced breast cancer therapy. The AHNP has been used as a defining tool to develop immunodetection probes that exemplify a general process application. AHNP has been expressed as an oligomeric fusion protein with streptavidin. These Herceptin like ABiPs were used to detect the Her2/neu antigen at extremely low concentrations using the immunodetection amplification technique (IDAT) which our laboratory has also developed. A fully developed highly diverse library of ABiPs represents an alternative for panels of monoclonal antibodies and may also be useful for target validation, antigen detection, therapeutics and as a platform for drug development. PMID- 12887104 TI - Immunoregulation of cellular lifespan: physiologic autoantibodies and their peptide antigens. AB - Physiologic autoantibodies are part of our normal immune repertoire where they function to maintain homeostasis by performing physiologic functions. The role of physiologic autoantibodies in removing senescent and damaged cells is probably the best example of a physiologic autoantibody, complete with well established function. IgG autoantibodies bind to altered band 3 anion exchanger protein on senescent cells and trigger their removal by macrophages. Band 3 isoforms are found in all cells, tissues, and membranes, and in all species examined. In this paper, we discuss the innate immune response to band 3 membrane proteins and their regulation of cellular lifespan. The role of physiologic autoantibodies and their peptide antigens in health and disease, apoptosis, and their therapeutic potential is discussed focusing on the examples of senescence and malaria. PMID- 12887105 TI - Peptide mimotopes as prototypic templates of broad-spectrum surrogates of carbohydrate antigens. AB - Peptide mimetics of carbohydrate antigens can function as templates to exploit immune mechanisms to augment vaccine design strategies as they are T cell dependent antigens. In this study we evaluate a peptide mimetic (peptide 105) of the Pneumococcal capsular polysaccharide type 14 (Pn14) as a model antigen to explore differences in antigenicity and immunogenicity of peptide mimotopes. The multiple antigenic peptide (MAP) form, by ELISA, competes with native Pn14 in a concentration-dependent manner for binding to an anti-Pn14 monoclonal antibody. It was observed that peptide priming with a conjugated form (105-BSA) and boosting with Pn14 produced higher levels of Pn14-reactive IgG1, IgG2a, IgG2b and IgG3 than priming and boosting with Pn14. This serum also displayed reactivity with multiple serotypes, as assessed by ELISA. However, when compared with serum from humans immunized with the 23-valent pneumococcal vaccines, mimetic-induced mouse serum did not display a significant ability to mediate opsonophagocytic killing of pneumococci. These results suggest the feasibility of designing mimotopes to render effective humoral responses not only to a single serotype of Streptococcus pneumoniae, but to multiple serotypes at once. Such peptides would simplify currently available vaccine approaches, yet highlights the requirement of more extensive polymerization to fully emulate native antigen. PMID- 12887106 TI - Using peptide mimetopes to elucidate anti-polysaccharide and anti-nucleic acid humoral responses. AB - Humoral responses against polysaccharide or nucleic acid antigens are often difficult to characterize and to induce. For example, the eliciting antigen for the development of anti-double-stranded(ds)DNA antibodies is unclear. dsDNA is a poor immunogen, yet antibodies to it bear the hallmark of a T cell dependent response. The microbial origin of polysaccharide antigens is, in general, readily known, but these antigens often do not elicit B cell memory responses, which are crucial for vaccine development. This review focuses on the use of peptide mimetopes to better understand humoral responses against non-protein antigens. First we describe a mimetope for dsDNA that was derived by probing a peptide phage library with an anti-dsDNA antibody. We discuss the usefulness of this mimetope in a search for candidate protein antigens and for examining the phenotype of antigen-specific B cells. Next, we discuss two mimetopes for phosphorylcholine (PC), a component of S. pneumoniae C polysaccharide. One was derived through mapping an anti-idiotype epitope and the other by probing a phagodisplay peptide library with an anti-PC antibody. Both of these peptide mimetopes for PC provide useful information regarding the requirements of a protective antibody response against pneumococcal infection, and define a critical role for adjuvant and carrier as well as mimetope. PMID- 12887107 TI - Evolution of the immunomodulatory role of the heat shock protein gp96. AB - In mammals, certain heat shock proteins (hsps) participate in specialized responses to stressors associated with pathogens or tumors, and as such, act as agents of immune surveillance interacting with both innate and adaptive immunity. We are investigating the conservation of this role throughout the class of vertebrates. We have shown that in Xenopus as in mammals, gp96, the major resident of the endoplasmic reticulum, generates MHC-restricted thymus-dependent immunity in vivo and CR in vitro against minor histocompatibility (H) antigens. By as yet unclear mechanisms that may involve classical MHC-unrestricted cytotoxic CD8+ T cells, gp96 also elicits peptide-specific responses against MHC class I-negative tumors in adult frogs that may involve cytotoxic NK, MHC unrestricted CD8+ T and NK/T cells. In naturally MHC class I-deficient but immunocompetent Xenopus larvae, gp96 also generates an innate type of anti-tumor response that is independent of chaperoned peptides. Finally, in a subset of Xenopus sIgM+ B cells, a substantial fraction of gp96 is directed to the cell surface by an active process that is upregulated by bacterial stimulation. This may allow gp96 to access the extracellular compartment without necrosis. Given the dual abilities of gp96 to chaperone antigenic peptides and to modulate innate immune responses, we propose that stimulated B cells that are up-regulating surface gp96 can directly interact with antigen presenting cells (APC) and/or T helper (Th) cells to trigger or amplify immune responses. PMID- 12887108 TI - The immunological functions of the vitamin D endocrine system. AB - The discoveries that activated macrophages produce 1alpha25-dihydroxyvitamin D3 (1alpha,25-(OH)2D3), and that immune system cells express the vitamin D receptor (VDR), suggested that the vitamin D endocrine system influences immune system function. In this review, we compare and contrast how 1alpha,25-(OH)2D3 synthesis and degradation is regulated in kidney cells and activated macrophages, summarize data on hormone receptor function and expression in lymphocytes and myeloid lineage cells, and discuss how locally-produced 1alpha,25-(OH)2D3 may activate a negative feed-back loop at sites of inflammation. Studies of immunity in humans and animals lacking VDR function, or lacking vitamin D, are viewed to gain insight into the immunological functions of the vitamin D endocrine system. The strong associations between poor vitamin D nutrition, particular VDR alleles, and susceptibility to chronic mycobacterial infections, together with evidence that 1alpha,25-(OH)2D3 served as a vaccine adjuvant enhancing antibody-mediated immunity, suggest a model wherein high levels of 1alpha,25-(OH)2D3-liganded VDR transcriptional activity may promote the CD4+ T helper 2 (Th2) cell-mediated and mucosal antibody responses to cutaneous antigens in vivo. We also review a diverse and rapidly growing body of epidemiological, climatological, genetic, nutritional and biological evidence indicating that the vitamin D endocrine system functions in the establishment and/or maintenance of immunological self tolerance. Studies done in animal models of multiple sclerosis (MS), insulin dependent diabetes mellitus (IDDM), inflammatory bowel disease (IBD), and transplantation support a model wherein the 1alpha,25-(OH)2D3 may augment the function of suppressor T cells that maintain self tolerance to organ-specific self antigens. The recent progress in infectious disease, autoimmunity and transplantation has stimulated a gratifying renaissance of interest in the vitamin D endocrine system and its role in immunological health. PMID- 12887109 TI - Evidence for structurally conserved recognition of the major carbohydrate xenoantigen by natural antibodies. AB - Natural or preformed antibodies that react with oligosaccharides bearing terminal galactose-alpha(1,3)-galactose [Gal alpha(1,3)Gal] stuctures are present in the sera of all humans. Antibodies against Gal alpha(1,3)Gal epitopes initiate hyperacute rejection of xenografts of porcine organs in human recipients. Despite the enormous clinical potential for xenotransplantation, very little is known about the 3D structural basis for natural antibody recognition of the major xenoantigen (i.e. Gal alpha(1,3)Gal). In this review, we discuss general binding patterns that have been repeatedly identified in antibody complexes with small molecules (haptens), carbohydrate and peptide ligands because similar mechanisms will almost certainly mediate recognition of the major xenoantigen by natural antibodies. PMID- 12887110 TI - Eating disorders: lifestyle choice or clinical problem? PMID- 12887111 TI - ACE inhibitor use and elevated creatinine levels. PMID- 12887112 TI - Potential causes of secondary hypertension. PMID- 12887113 TI - Prescribing syringes to injection drug users: what the family physician should know. PMID- 12887114 TI - Diagnosing acute monoarthritis in adults: a practical approach for the family physician. AB - Acute monoarthritis can be the initial manifestation of many joint disorders. The first step in diagnosis is to verify that the source of pain is the joint, not the surrounding soft tissues. The most common causes of monoarthritis are crystals (i.e., gout and pseudogout), trauma, and infection. A careful history and physical examination are important because diagnostic studies frequently are only supportive. Examination of joint fluid often is essential in making a definitive diagnosis. Leukocyte counts vary widely in septic and sterile synovial fluids and should be interpreted cautiously. If the history and diagnostic studies suggest an infection, aggressive treatment can prevent rapid joint destruction. When an infection is suspected, culture and Gram staining should be performed and antibiotics should be started. Light microscopy may be useful to identify gout crystals, but polarized microscopy is preferred. Blood tests alone never confirm a diagnosis, and radiographic studies are diagnostic only in selected conditions. Referral is indicated when patients have septic arthritis or when the initial evaluation does not determine the etiology. PMID- 12887115 TI - Common hair loss disorders. AB - Hair loss (alopecia) affects men and women of all ages and often significantly affects social and psychologic well-being. Although alopecia has several causes, a careful history, dose attention to the appearance of the hair loss, and a few simple studies can quickly narrow the potential diagnoses. Androgenetic alopecia, one of the most common forms of hair loss, usually has a specific pattern of temporal-frontal loss in men and central thinning in women. The U.S. Food and Drug Administration has approved topical minoxidil to treat men and women, with the addition of finasteride for men. Telogen effluvium is characterized by the loss of "handfuls" of hair, often following emotional or physical stressors. Alopecia areata, trichotillomania, traction alopecia, and tinea capitis have unique features on examination that aid in diagnosis. Treatment for these disorders and telogen effluvium focuses on resolution of the underlying cause. PMID- 12887116 TI - Information from your family doctor. What should I know about hair loss? PMID- 12887117 TI - Black cohosh. AB - The herb black cohosh, or Actaea racemosa (formerly named Cimicifuga racemosa), is native to North America. The roots and rhizomes of this herb are widely used in the treatment of menopausal symptoms and menstrual dysfunction. Studies have demonstrated that this botanic medicine, when standardized properly to the terpene glycoside fraction, appears to be effective in alleviating menopausal symptoms. Adverse effects are extremely uncommon, and there are no known significant adverse drug interactions. PMID- 12887118 TI - Nausea and vomiting of pregnancy. AB - Nausea and vomiting of pregnancy, commonly known as "morning sickness," affects approximately 80 percent of pregnant women. Although several theories have been proposed, the exact cause remains unclear. Recent research has implicated Helicobacter pylori as one possible cause. Nausea and vomiting of pregnancy is generally a mild, self-limited condition that may be controlled with conservative measures. A small percentage of pregnant women have a more profound course, with the most severe form being hyperemesis gravidarum. Unlike morning sickness, hyperemesis gravidarum may have negative implications for maternal and fetal health. Physicians should carefully evaluate patients with nonresolving or worsening symptoms to rule out the most common pregnancy-related and nonpregnancy related causes of severe vomiting. Once pathologic causes have been ruled out, treatment is individualized. Initial treatment should be conservative and should involve dietary changes, emotional support, and perhaps alternative therapy such as ginger or acupressure. Women with more complicated nausea and vomiting of pregnancy also may need pharmacologic therapy. Several medications, including pyridoxine and doxylamine, have been shown to be safe and effective treatments. Pregnant women who have severe vomiting may require hospitalization, orally or intravenously administered corticosteroid therapy, and total parenteral nutrition. PMID- 12887119 TI - Management of Bartholin's duct cyst and gland abscess. AB - Bartholin's duct cysts and gland abscesses are common problems in women of reproductive age. Bartholin's glands are located bilaterally at the posterior introitus and drain through ducts that empty into the vestibule at approximately the 4 o'clock and 8 o'clock positions. These normally pea-sized glands are palpable only if the duct becomes cystic or a gland abscess develops. The differential diagnosis includes cystic and solid lesions of the vulva, such as epidermal inclusion cyst, Skene's duct cyst, hidradenoma papilliferum, and lipoma. The goal of management is to preserve the gland and its function if possible. Office-based procedures include insertion of a Word catheter for a duct cyst or gland abscess, and marsupialization of a cyst; marsupialization should not be used to treat a gland abscess. Broad-spectrum antibiotic therapy is warranted only when cellulitis is present. Excisional biopsy is reserved for use in ruling out adenocarcinoma in menopausal or perimenopausal women with an irregular, nodular Bartholin's gland mass. PMID- 12887120 TI - Nausea and vomiting in early pregnancy. PMID- 12887121 TI - Behavioral counseling in primary care to promote a healthy diet. PMID- 12887122 TI - Photo quiz. Obesity and daytime sleepiness. Prader-Willi syndrome. PMID- 12887123 TI - NAEPP updates guidelines for the diagnosis and management of asthma. PMID- 12887124 TI - Disability revisited. PMID- 12887125 TI - Renin, angiotensin, sodium and organ damage. AB - Angiotensin II and sodium balance affect the status of each other and both- either separately or together--can lead to an increase in blood pressure. They also can cause vascular and cardiac damage due to direct effects and effects mediated by the blood pressure increase. This paper reviews the important interactions among these three variables. Acute blood pressure elevation during sleeping but not during waking hours causes cardiac hypertrophy in rats. Similarly, lowering of blood pressure with an angiotension converting enzyme (ACE) inhibitor during sleep but not when awake causes regression of cardiac hypertrophy in rats with 2kidney (K)-1clip (C) Goldblatt hypertension. If angiotensin is given to rats on a low (0.4%) NaCl diet, blood pressure rises but there is less cardiac hypertrophy. Cardiac hypertrophy is greatest after angiotensin administration in rats on a high (4%) NaCl diet. In both the 2K-1C and 1K-1C Goldblatt models, a high salt intake reduces the blood pressure lowering effect of captopril and losartan and prevents regression of cardiac hypertrophy. Combined administration of an ACE inhibitor and an angiotensin type 1 (AT1) receptor blocker to rats on a low (0.2%) NaCl diet produces a syndrome that leads to death with cardiac involution. All features of the syndrome are reversed or prevented by 4% NaCl intake. It is hypothesised that the interaction between angiotensin II and sodium intake can be explained by differences in the way protons produced by contracting cells are neutralized. The sodium hydrogen exchanger and the sodium 2 bicarbonate cotransporter are stimulated by the AT1 and angiotensin type 2 (AT2) receptor, respectively. If the ratio of receptors is altered in favour of the AT2 receptor, then less cardiac hypertrophy will result from the same workload. Review of the clinical literature reveals that many of these results in rats have correlations in clinical medicine. Thus high night time blood pressure is associated with a greater morbidity and high salt intake causes cardiac hypertrophy and vascular stiffness independent of blood pressure levels. When deciding on treatment in human hypertension these results have important clinical implications. PMID- 12887126 TI - Disasters and the heart: a review of the effects of earthquake-induced stress on cardiovascular disease. AB - There is growing evidence that stress contributes to cardiovascular disease. Chronic stress contributes to the atherosclerotic process through increased allostatic load, which is mediated by the neuroendocrine and immune systems (sympathetic nervous system and hypothalamus-pituitary adrenal axis) and related chronic risk factors (insulin resistance syndrome, hypertension, diabetes, and hyperlipidemia). In addition, acute stress can trigger cardiovascular events predominantly through sympathetic nervous activation and potentiation of acute risk factors (blood pressure increase, endothelial cell dysfunction, increased blood viscosity, and platelet and hemostatic activation). Earthquakes provide a good example of naturally occurring acute and chronic stress, and in this review we focus mainly on the effects of the Hanshin-Awaji earthquake on the cardiovascular system. The Hanshin-Awaji earthquake resulted in a 3-fold increase of myocardial infarctions in people living close to the epicenter, particularly in women, with most of the increase occurring in nighttime-onset events. There was also a near doubling in the frequency of strokes. These effects may be mediated by changes in hemostatic factors, as demonstrated by an increase of D dimer, von Willebrand factor, and tissue-type plasminogen activator (tPA) antigen. Blood pressure also increased after the earthquake, and was prolonged for several weeks in patients with microalbuminuria. PMID- 12887127 TI - Effects of hormone replacement therapy on office and ambulatory blood pressure in Japanese hypertensive postmenopausal women. AB - No study has demonstrated that hormone replacement therapy (HRT) affects blood pressure (BP) measured by 24-h ambulatory blood pressure monitoring (ABPM) in Japanese postmenopausal women (PMW) with normotension or mild-to-moderate essential hypertension. In the present study, we examined the effects of HRT on office BP and 24-h ambulatory blood pressure (ABP) in Japanese hypertensive or normotensive PMW. Thirty-one hypertensive (HT-HRT group) and 17 normotensive PMW (NT-HRT group) received HRT (0.625 mg of conjugated equine estrogen combined with 2.5 mg of medroxyprogesterone acetate) orally for 12 months, and 30 hypertensive (HT-Control group) and 19 normotensive PMW (NT-Control group) did not receive HRT. In all of the hypertensive PMW, BP was controlled by a variety of antihypertensive drugs before starting HRT. The hypertensive PMW were divided into two groups according to the results of ABP before HRT: nondippers (those without a diurnal change in BP) and dippers (those with a diurnal change in BP). In all patients, office BP measurements and 24-h ABPM were performed before and 12 months after the start of HRT. HRT did not change either the office or the 24 h ambulatory systolic, diastolic, or mean BP in any of the groups. Therefore, HRT did not significantly alter the proportion of nondippers. We conclude that with respect to BP, HRT might not be harmful in hypertensive PMW whose BP has been well-controlled prior to the initiation of HRT, as well as in normotensive PMW. PMID- 12887128 TI - Ankle-arm blood pressure index and cardiovascular risk factors in elderly Japanese men. AB - Low ankle-arm systolic blood pressure index (AAI) correlates with various cardiovascular risk factors and with risk of subsequent coronary heart disease and stroke in Western countries. However, few epidemiological data are available among Japanese, in whom the reported prevalence of peripheral artery disease is low. We examined the relationship between AAI and cardiovascular risk factors among 1,219 men aged 60 to 79 years in two Japanese communities in 1999 and 2000. The prevalence of AAI<0.90 was 5% in both communities. Hypertension, major ECG abnormality, current smoking, and history of stroke were associated with two- to four-fold higher prevalence of AAI<0.90. One-standard deviation increments of body mass index and high density lipoprotein-cholesterol levels were associated with 60% and 40% lower prevalence of AAI<0.90, respectively. Although the prevalence of low AAI in Japanese elderly men is lower than that reported in the Unites States and European studies, similar correlations of low AAI with cardiovascular risk factors were observed among different ethnic groups. Low AAI is suggested to be a predictor for stroke among Japanese men, which should be confirmed by a prospective study. PMID- 12887129 TI - Effect of cilnidipine on insulin sensitivity in patients with essential hypertension. AB - To clarify the effect of cilnidipine, a long-acting dihydropyridine Ca-antagonist that blocks both L- and N-type Ca(2+)-channels, on insulin sensitivity, cilnidipine at 5 to 10 mg/day was administered to ten patients with essential hypertension for 12 weeks. Mean age and body mass index (BMI) were 57.7 +/- 5.0 (SEM) years old and 27.1 +/- 1.5, respectively. Blood pressure, serum levels of catecholamines, glucose and lipid were determined before and after the treatment. Insulin sensitivity was also measured by a euglycemic hyperinsulinemic clamp method using an artificial pancreas (STG-22; Nikiso, Tokyo, Japan) before and after the treatment. Cilnidipine administration significantly lowered blood pressure from 154/96 to 137/84 mmHg (p<0.05). The glucose infusion rate was significantly increased by 20.8%, from 3.27 +/- 0.36 to 3.95 +/- 0.55 mg/kg/min (p<0.05). HbA1C and serum lipid levels such as total cholesterol and triglyceride were not altered. In addition, cilnidipine treatment did not significantly increase serum norepinephrine levels (278 +/- 25.2 vs. 332 +/- 33.6 pg/ml). Our results suggest that cilnidipine improves insulin sensitivity, possibly due to its exerting a vasodilatory action without stimulating sympathetic nervous activity. PMID- 12887130 TI - The effect of tumor necrosis factor-alpha on tissue specificity and selectivity to insulin signaling. AB - Recent studies have indicated that tumor necrosis factor (TNF)-alpha plays a significant role in insulin resistance. It has been proposed that selective impairment of insulin signaling in glucose metabolism is related to the development of atherosclerosis, although the mechanisms are not clear. The aim of this study was to elucidate the effect of TNF-alpha on tissue specificity and selectivity to insulin signaling. L6 myotubes and rat aortic vascular smooth muscle cells (VSMC) were cultured. Cells were stimulated with insulin pretreated with or without TNF-alpha. The protein extracts were used for electrophoresis and immunoblotting studies to examine phosphorylation of insulin receptor (IR)-beta, insulin receptor substrate (IRS)-1 and extracellular signal-regulated kinase (ERK). IR-beta phosphorylation was not affected by TNF-alpha in L6 or in VSMC. TNF-alpha significantly (p<0.05) inhibited IRS-1 phosphorylation by insulin but had no effect on ERK in L6. TNF-alpha had no effect on either IRS-1 phosphorylation or ERK in VSMC. Insulin induced ERK phosphorylation in a dose dependent manner in VSMC. These results suggests that TNF-alpha plays a significant role in the tissue specificity and signal selectivity of insulin resistance. The pathway related to glucose metabolism is selectively impaired by TNF-alpha in skeletal muscle, and this impairment may induce compensatory hyperinsulinemia, which in turn would stimulate the pathway related to the cell proliferation in vascular tissues and possibly enhance the progression of atherosclerosis. PMID- 12887131 TI - Membrane ion transport in erythrocytes of salt hypertensive Dahl rats and their F2 hybrids: the importance of cholesterol. AB - The possible association of salt hypertension and altered lipid metabolism with abnormalities of particular systems transporting sodium and potassium has been studied in erythrocytes of Dahl rats and their F2 hybrids fed a high-salt diet since weaning. Our attention was paid to the Na(+)-K+ pump, Na(+)-K+ cotransport and especially to passive membrane permeability for Na+ and Rb+ (Na+ and Rb+ leak), because the Na+ leak was found to be dependent on the genotype, age and salt intake of Dahl rats, whereas the Rb+ leak was suggested to be a potential marker of salt sensitivity in Dahl and Sabra rats. Young male Dahl salt-sensitive (SS/Jr) and salt-resistant (SR/Jr) rats kept on a low-salt (0.3% NaCl) or high salt diet (8% NaCl) were used for the progenitor study. The subsequent genetic study was based on 135 young male SS/Jr x SR/Jr F2 hybrids fed a high-salt diet since weaning. Ouabain (5 mmol/l) and bumetanide (10 micromol/l) were used to distinguish the contribution of the Na(+)-K+ pump, Na(+)-K+ cotransport and passive membrane permeability to measured net Na+ fluxes and unidirectional Rb+ (K+) movements. Compared to normotensive SR/Jr animals, salt-loaded SS/Jr rats had higher blood pressure (BP), elevated erythrocyte Na+ content, and increased Na+ and Rb+ leaks together with enhanced Na+ and Rb+ transport mediated by the Na(+)-K+ pump and Na(+)-K+ cotransport system. Salt hypertensive Dahl rats were also characterized by elevated plasma levels of total cholesterol and triglycerides, which were positively associated with BP of F2 hybrids (r=0.27 and 0.24, p< 0.01). In F2 hybrids, mean arterial pressure correlated significantly with erythrocyte Na+ content (r=0.24, p<0.01) and ouabain-sensitive Na+ extrusion, but not with the passive membrane permeability for Na+ or Rb+ (r=-0.02 and 0.06, not significant). Both of the above-mentioned significant associations could partially be ascribed to the dependence of erythrocyte Na+ content and ouabain-sensitive Na+ extrusion on plasma cholesterol (r=0.18 and 0.21, p<0.05). Our results support the idea that abnormal lipid metabolism and/or altered Na+,K(+)-ATPase function play an important role in the pathogenesis of salt hypertension in salt-sensitive Dahl rats. PMID- 12887132 TI - Type 1A angiotensin II receptor is regulated differently in proximal and distal nephron segments. AB - Angiotensin II plays important roles in renal vasoconstriction, sodium reabsorption in proximal tubules, and cell proliferation. Angiotensin II receptors are present not only in proximal but also in distal tubules. We investigated the effects of dehydration on the mRNA expression of type 1A angiotensin II receptor (AT1A) in proximal and distal nephron segments and on the expression of type 1 angiotensin II receptor (AT1) protein. Competitive polymerase chain reaction was employed to quantitatively examine mRNA expression, and AT1-specific polyclonal antibody was used for Western blot analysis. AT1A mRNA expression was most abundant in glomeruli. Collecting ducts showed higher expressions than did proximal tubules or thick ascending limbs. Dehydration caused an increase of AT1A mRNA expression in glomeruli, proximal straight tubules (PST), and medullary and cortical thick ascending limbs (MAL and CAL, respectively). In contrast, dehydration decreased AT1A mRNA expression in cortical, outer medullary, and inner medullary collecting ducts (CCD, OMCD, and IMCD, respectively). Incubation of isolated glomeruli, PST, and IMCD in hypertonic solution made by NaCl and mannitol in vitro increased AT1A mRNA expression. Incubation of IMCD with AVP (10(-7) mol/l) also increased AT1A mRNA expression. AT1 was detected at 45 kDa by Western blotting. Dehydration caused a decrease and increase of AT1 expression in the cortex and the medulla, respectively. In summary, these data showed that the mechanisms of the regulation of AT1A differ between proximal and distal tubules. The finding that AT1 was up regulated in the medulla during dehydration may suggest that this receptor plays an important role in dehydration in the distal tubules. PMID- 12887133 TI - Angiotensin II blockade restores albumin reabsorption in the proximal tubules of diabetic rats. AB - The kidney plays an important role in protein metabolism. The albumin reabsorption in the proximal tubule is disturbed in the early stage of diabetic nephropathy. We evaluated the effects of angiotensin converting enzyme inhibitor (ACEI) and angiotensin III type 1 receptor blocker (ARB) on albumin reabsorption and expression of megalin, an endocytosis receptor for albumin, in proximal tubules of streptozotocin (STZ)-induced diabetic-rats. Diabetic rats at the second week after STZ injection were treated with quinapril (3 mg/kg/day) or candesartan (0.05 mg/kg/day) for 2 weeks. The tubular reabsorption of fluorescein isothiocyanate (FITC)-labeled albumin was evaluated by immunogold electron microscopy, and megalin expression was investigated by immunohistochemistry and Western blotting. Reabsorption of FITC-labeled albumin and megalin expression were prominently inhibited in the proximal convoluted tubules of diabetic rats compared to the controls. Both quinapril and candesartan restored albumin reabsorption in the proximal tubule due to normalization of megalin expression. Urinary albumin excretion was significantly reduced by both ACEI and ARB treatment. Angiotensin II infusion decreased megalin expression and albumin reabsorption in the proximal tubule. In conclusion, angiotensin II blockade restored albumin reabsorption via amelioration of megalin expression in the proximal tubules of early stage diabetic rats. PMID- 12887134 TI - Differential induction of protein kinase C isoforms at the cardiac hypertrophy stage and congestive heart failure stage in Dahl salt-sensitive rats. AB - Several protein kinase C (PKC) isoforms may play important roles in cellular signaling pathways. Recent reports have suggested that PKC plays critical isoform specific roles in the development of cardiac hypertrophy and heart failure. The purpose of the present study was to examine the expression profiles of PKC isoforms in models of cardiac hypertrophy and heart failure. We examined the cardiac expression of individual PKC isoforms at the cardiac hypertrophy stage and the heart failure stage in Dahl salt-sensitive rats by Western blot analysis. The levels of all PKC isoforms increased at the cardiac hypertrophy stage and the heart failure stage, but the pattern of increase differed among PKC isoforms at the heart failure stage. The expressions of PKCalpha, beta, and delta increased at the cardiac hypertrophy stage and remained elevated at the heart failure stage. On the other hand, the expression of PKCepsilon and atypical PKCs (aPKCs) increased at the cardiac hypertrophy stage, but this increase tended to decline at the congestive heart failure stage. These results suggest that there are two groups of PKC isoforms. Several reports have shown that PKCalpha, beta, and delta are involved in the development of cardiac hypertrophy and heart failure, and that PKCepsilon plays a role in the physiological hypertrophic responses and cardioprotective actions. These facts suggest that all PKC isoforms (PKCalpha, beta, delta, epsilon, and aPKCs) expressed in the heart may have similar functions at the cardiac hypertrophy stage, but that two groups of PKC isoforms (PKCalpha, beta, delta, and PKCepsilon, aPKCs) have different functions at the congestive heart failure stage. PMID- 12887135 TI - Lipopolysaccharide reverses adrenocorticotrophic hormone-induced hypertension in the rat. AB - Lipopolysaccharide (LPS) was used to stimulate nitric oxide (NO) release and investigate the effect of endogenous NO on adrenocorticotrophic hormone (ACTH) induced hypertension in rats. After preliminary studies to determine the appropriate dose of LPS, 40 male Sprague-Dawley rats were treated with ACTH (200 microg/kg/day, s.c.) or saline (sham) for 8 days and then given a single dose of LPS (10 mg/kg, i.p.) or saline. ACTH treatment was continued for a further 5 days. Systolic blood pressure (SBP) was measured daily using the tail cuff method. Results were expressed as the mean +/- SEM. ACTH treatment significantly increased SBP (from 105 +/- 3 to 129 +/- 4 mmHg; p<0.05), whereas saline had no effect on SBP. The ACTH-induced increase in SBP was reversed by LPS injection (from 125 +/- 6 to 102 +/- 7 mmHg; p<0.05). SBP was also decreased in sham + LPS treated rats compared with that of sham + saline-treated rats (p<0.05), but the SBP change in response to LPS was greater in ACTH-treated than in sham-treated rats (-23 vs. -8 mmHg; p<0.05). These data are compatible with the notion that reduced NO availability plays a role in ACTH-induced hypertension. PMID- 12887136 TI - Two cases of pheochromocytoma associated with tetralogy of Fallot. AB - We report two cases of pheochromocytoma combined with tetralogy of Fallot who showed different clinical courses. Case 1 was a 45-year-old woman with a history of radical operation for tetralogy of Fallot at 20 years of age. She presented with sudden hypertensive attack, and was diagnosed with pheochromocytoma of the left adrenal gland. She was treated surgically, and her high plasma noradrenaline level normalized. Case 2 was a 41-year-old woman who had been suffering from severe cyanosis due to tetralogy of Fallot throughout her life. A palliative operation had been performed at 7 years of age, but a radical operation had not been performed. She has had resistant hypertension since 38 years of age. She was diagnosed as having pheochromocytoma of the left adrenal gland at 41 years of age, but surgery was not performed. She was pharmacologically treated with doxazosin, followed by bisoprolol. Her symptoms somewhat improved, although she continued to have high plasma levels of noradrenaline and adrenomedullin. The combination of pheochromocytoma with tetralogy of Fallot or cyanotic congenital heart disease is rare; however, pheochromocytoma and congenital heart disease might be related through chronic hypoxia and/or gene abnormalities. The presence of pheochromocytoma worsens the hemodynamic state in patients with congenital heart disease regardless of whether radical surgery for congenital heart disease had been performed. Differential diagnosis of pheochromocytoma could be paramount in congenital heart disease patients who show unexpected or unusual symptoms. PMID- 12887137 TI - Embryonic exposure to exogenous alpha- and gamma-tocopherol partially attenuates ethanol-induced changes in brain morphology and brain membrane fatty acid composition. AB - Previous studies demonstrated that embryonic exposure to ethanol (EtOH) promoted a reduction in brain mass, a reduction in brain neuron densities, and a reduction in membrane long-chain polyunsaturated fatty acids (PUFAs) in embryonic chick brains. These EtOH-induced reductions in brain membrane PUFAs may be the result of lipid peroxidation because embryonic exposure to exogenous alpha- or gamma tocopherol partially attenuated EtOH-induced reductions in membrane PUFAs. In this paper, we report that embryonic exposure to exogenous alpha- or gamma tocopherol attenuated EtOH-induced decreases in endogenous levels of alpha tocopherol in both embryonic chick brains and liver. Embryonic exposure to exogenous alpha- or gamma-tocopherol also partially attenuated EtOH-induced reductions in brain neuron densities within the cerebral hemispheres of embryonic chick brains. Finally, embryonic exposure to exogenous alpha- or gamma-tocopherol also partially attenuated EtOH-induced reductions in long-chain PUFAs in 2-day old neonatal chick brains. PMID- 12887138 TI - Postnatal protein malnutrition affects inhibitory avoidance and risk assessment behaviors in two models of anxiety in rats. AB - Protein malnutrition induces structural, neurochemical and functional alterations in the central nervous system, leading to alterations in behavioral function. In order to study the effects of early protein malnutrition on inhibitory avoidance and escape behaviors we used the elevated T-maze (ETM), while the risk assessment behaviors were evaluated by the canopy stretched attend posture (SAP) test. Rat pups were fed by lactating females receiving 16% (control) or 6% (malnourished) protein diets during the lactation period. After weaning the animals received the same diets until 49 days of age, when all animals started receiving a lab chow diet. Behavioral tests were started at 70 days of age. ETM results showed lower inhibitory avoidance in malnourished animals, without differences in escape behavior. SAP test results showed higher exploration and lower risk assessment behaviors in malnourished animals compared to control. These results suggest that malnourished animals are less anxious and/or more impulsive as measured by these two animal models and that malnutrition seems to affect differently behavioral strategies underlying fear and anxiety responses. PMID- 12887139 TI - A comparison of tyrosine against placebo, phentermine, caffeine, and D amphetamine during sleep deprivation. AB - Sleep deprivation can impair alertness and cognitive and motor performance. We hypothesized that the amino acid tyrosine might reduce deleterious effects of sleep deprivation. Seventy-six healthy males, age 18-35 years, participated in a four-day protocol that included a habituation night, a baseline night, a 40.5 h period without sleep, and a recovery night. Tyrosine 150 mg/kg, caffeine 300 mg/70 kg, phentermine 37.5 mg, D-amphetamine 20 mg and placebo were administered in a double-blind, randomized fashion to compare their effects on the time it took to fall asleep, on endocrine responses during sleep deprivation, and on sleep quantity, quality and architecture as measured by polysomnography during recovery sleep. When given after 36 h without sleep, tyrosine had no significant effect on any parameter of sleep. D-amphetamine produced marked decrease in sleep drive but caused deleterious effects on many aspects of recovery sleep. Still, D amphetamine was associated with increased alertness on the first recovery day. Phentermine and caffeine both decreased sleep drive during sleep deprivation, but phentermine impaired rapid-eye-movement (REM) recovery sleep. Tyrosine (when compared to placebo) had no effect on any sleep related measure, but it did stimulate prolactin release. PMID- 12887140 TI - Effects of tyrosine, phentermine, caffeine D-amphetamine, and placebo on cognitive and motor performance deficits during sleep deprivation. AB - Cognitive and motor performance are critical in many circumstances and are impaired by sleep deprivation. We administered placebo, tyrosine 150 mg/kg, caffeine 300 mg/70 kg, phentermine 37.5 mg and D-amphetamine 20 mg at 15.30 h following overnight sleep deprivation and compare their effects on cognitive and motor performance in healthy young men. Tests of visual scanning, running memory, logical reasoning, mathematical processing, the Stroop task, four-choice serial reaction time, time wall take, pursuit tracking, visual vigilance, Trails (B) task and long-term memory were evaluated at standardized intervals before, during and after sleep deprivation and drugs. Performance decrements with sleep deprivation occurred in visual scanning, running memory, logical reasoning, mathematical processing, the Stroop test, the time wall test, tracking and visual vigilance. Interestingly, with sleep deprivation some tests improved and others did not deteriorate. Improvements with medication following sleep deprivation were seen in running memory, logical reasoning, mathematical processing, tracking and visual vigilance. Although less effective than D-amphetamine, tyrosine improved performance on several tests. We conclude that all drugs tested improved at least some aspects of cognitive and motor performance after sleep deprivation. As a naturally occurring amino acid, and thus amenable to nutritional strategies, tyrosine may deserve further testing. PMID- 12887141 TI - The rapid anorectic response to a threonine imbalanced diet is decreased by injection of threonine into the anterior piriform cortex of rats. AB - Rats quickly recognize and reject diets deficient in an essential amino acid. The purpose of this study was to determine whether the anterior piriform cortex (APC), the site traditionally recognized as the amino acid chemosensor, plays a role in this early behavior. Rats had cannulae implanted bilaterally into the APC, and were injected with either saline vehicle or 2 nmoles of threonine (n = 6 per group). All rats were then fed a diet imbalanced with respect to threonine. The threonine-injected group had first meals of longer duration and consumed more food. These data conformed to expectations derived from earlier studies of responses to the first meal of an amino acid imbalanced diet. We conclude that the concentration of the dietary limiting amino acid in the APC regulates acceptance and rejection of amino acid deficient diets. PMID- 12887142 TI - Effects of taurine deficiency and chronic methanol administration on rat retina, optic nerve and brain amino acids and monoamines. AB - A chronic methanol (MeOH) intoxication scheme (2 g/kg/day ip for 2 weeks) was carried out in Sprague-Dawley rats, previously depleted of folates with methotrexate (MTX). beta-Alanine (beta-Ala), 5%, was also administered to some animals in the drinking water. Amino acids were determined in plasma, retina, optic nerve, hippocampus and posterior cortex by HPLC with fluorescence detection and monoamines in retina, hippocampus and posterior cortex by electrochemical detection. Beta-Ala administration reduced taurine (Tau) levels in plasma, hippocampus and posterior cortex, but not in retina and optic nerve. Aspartate (Asp) concentration in the optic nerve was increased in MTX-MeOH treated animals, and the administration of beta-Ala did not modify this elevation. The association of beta-Ala with MTX-MeOH produced an increase of threonine, and a decrease of 5 hydroxytryptamine (5-HT) in the retina without modifying 5-hydroxyindoleacetic acid, whereas in the hippocampus an elevation of asparagine was observed. We conclude that, in the retina, beta-Ala in combination with MTX-MeOH increased serotonin and decreased dopamine (DA) turnover rate, and resulted in changes in the amino acid balance, that could affect glycinergic activity. On the other hand, in the hippocampus, Asp metabolism could be affected by Tau depletion with beta-Ala. PMID- 12887143 TI - Prenatal protein malnutrition results in increased frequency of miniature inhibitory postsynaptic currents in rat CA3 interneurons. AB - Electrophysiological studies have revealed an increase in the level of tonic inhibition in the hippocampus following prenatal protein malnutrition in rats. In the present study, whole cell patch clamp recordings of bipolar interneurons in the stratum radiatum of the CA3 subfield were used to determine whether this increase in inhibition can be accounted for by a change in the electrophysiological properties of GABAergic interneurons. Hippocampal slices were prepared from juvenile rats whose dams were fed either a normal (25% casein) or low (6% casein) protein diet throughout pregnancy. Intrinsic membrane and action potential properties were unaltered by the prenatal nutritional insult. In most respects the characteristics of GABAA receptor-mediated inhibitory postsynaptic currents (IPSCs) and the modulation of such currents by the benzodiazepine agonist zolpidem were also similar in cells from the two nutritional groups. While the frequency of spontaneous inhibitory currents was unaltered, miniature (Tetrodotoxin resistant) inhibitory currents occurred at a significantly increased frequency in interneurons from prenatally protein malnourished rats. Thus, while the basic membrane properties of interneurons are preserved, there is a significant increase in the probability of GABA release from interneurons following prenatal protein malnutrition. PMID- 12887144 TI - Proliferative effect of growth factors TGF-beta1, PDGF-BB and rhBMP-2 on human gingival fibroblasts and periodontal ligament cells. AB - The regeneration of periodontal tissues lost due to periodontal disease requires cell migration, differentiation and proliferation. Several procedures have been proposed to promote wound healing events such as the application of growth factors including PDGF-BB, TGF-beta1 and rhBMP-2. The purpose of this study was to evaluate the mitogenic responses of human periodontal ligament cells and gingival fibroblasts to PDGF-BB, TGF-beta1 and rhBMP-2. Human periodontal ligament cells were isolated from the mid root of three maxillary third molars extracted from three adult patients with moderate periodontitis and gingival fibroblasts were obtained from two patients also affected by moderate periodontitis, who underwent periodontal surgery. Cells were grown in 24-well dishes. On day 2 of quiescence, new medium was added with PDGF-BB or TGF-beta1 or rhBMP-2 at the concentration of 10 ng/ml. To determine the effects of the test agents on cell proliferation, DNA synthesis was estimated by measuring [3H] thymidine incorporation. After 48h of incubation the cells were processed and subject to scintillation counting. Counts per minute (cpm/ well) were determined for each sample. The results of this study demonstrated that PDGF-BB acts like a strong mitogenic agent for human periodontal ligament cells and gingival fibroblasts, TGF-beta1 mostly supports the proliferation of these cells and rhBMP 2 had an opposite effect on cell mitosis. PMID- 12887145 TI - Reliability of bleeding and non-bleeding on probing to gingival histological features. AB - Given the extensive use in periodontal diagnosis of the clinical sign of bleeding on probing, the reliability of the technique in relation to gingival histological features and the need for a better understanding of its significance was investigated. Bleeding (B) and non-bleeding (NB) buccal gingival tissues, were measured through manual probing during gingivectomy procedures. The histological evidence of inflammation was used as the gold standard. The established histological descriptive patterns showed statistically significant differences between B and NB sites (chi2 = 20.842, p < 0.05). The reliability scores were found to be of high sensitivity (90.9%) and negative predictive value (89.5%) as well as of fair specificity (77.3%) and positive predictive value (80%). The accuracy was found to be 84.1%. The histological quantitative analysis showed no statistically significant difference for any HGI (Histological Gingival Index). B sites presented HGI = 1 in 72.8%, close to the NB sites, which showed 63.4% for the same histological score. Furthermore, Pearson's analysis showed a low correlation between the studied clinical signs and the histological quantitative analysis (r = 0.265, p > 0.05). We conclude that the clinical parameter of bleeding on probing is a reliable tool for diagnosing gingival inflammation, whereas the clinical sign of non-bleeding on probing is not a safe criteria for evaluating gingival health. PMID- 12887146 TI - Periodontal disease among the middle-aged Vietnamese population. AB - Information about periodontal disease patterns among the populations of developing countries is limited. These populations may illustrate the natural history of the disease owing to low or virtually no access to dental care, contributing to a more complete understanding of the disease process. The present study aimed to describe the patterns of periodontal loss of attachment (LOA) among representative middle-aged Vietnamese adults. A multi-stage, stratified random sampling technique was employed in two randomly selected provinces. The US NIDR protocol with two sites per tooth for the whole mouth, excluding third molars, was used to assess LOA. A total of 575 subjects aged 35-44 years were interviewed and periodontally examined (response 84.6%). Data were re-weighted to reflect the population of the two provinces. Oral hygiene status was poor with high accumulation of plaque and calculus. The prevalence, extent of sites with pockets (PD), recession (GR) and LOA exceeding various thresholds and the severity of LOA were calculated. Almost all subjects presented with at least one site with LOA > or = 2 mm but the higher levels of the disease were skewed. The extent of sites with GR, PD and LOA > or = 4mm was 1.35, 5.04 and 11.81 per cent respectively. More buccal sites had GR, while more mesial sites had PD and LOA. Severity score of LOA was 2.5 (SD: 0.7). LOA of 4+mm were observed more at maxillary molar buccal sites, followed by mandibular incisor mesial sites. LOA was prevalent among middle-aged Vietnamese adults. The loss of periodontal support was positively skewed and varied between teeth and sites. PMID- 12887147 TI - Philosophical approach in old adults dental treatment. PMID- 12887148 TI - Oral diseases and conditions in community-living older adults with and without dementia. AB - This study compared the prevalence and experiences of oral diseases and conditions, as well as tooth loss and retained root status, in community-dwelling older adults with and without dementia. Dental examinations were conducted at baseline and at one-year for randomly selected samples of 116 dentate community living older adults with dementia and a comparable group of 116 adults without dementia. Participants with dementia had significantly higher experiences of oral diseases and conditions at baseline and one-year compared with participants without dementia: decreased use of dentures; increased prevalence of denture related oral mucosal lesions; increased plaque accumulation; increased prevalence and experiences of coronal and root caries; and increased numbers of decayed retained tooth roots. These higher experiences of oral diseases and conditions were related to dementia severity, not to specific dementia diagnoses. Participants with dementia already had a compromised oral health status when admitted into institutional long-term care between baseline and one-year; of concern were the high plaque levels on the natural teeth of the group of institutionalized participants with dementia. PMID- 12887149 TI - Explorer probing of root caries lesions: an in vitro study. AB - Probing with a dental explorer is the preferred method of root caries diagnoses. However, studies suggest that use of a dental explorer in coronal caries detection may damage tooth structure. This pilot study investigated the effect of probing on remineralization of root surface lesions in vitro. Root-surface lesions were created on 10 extracted teeth by exposing them to a demineralizing solution for 21 days. One side of each lesion was then randomly selected and probed with an explorer, while the other side was not probed. The teeth then were placed in a remineralizing solution containing 10 parts per million (ppm) fluoride for 21 days. Of the 9 teeth with usable sections, all showed frank evidence of defects on the probed halves. Sites of remineralization were seen in the non-probed halves and adjacent to the probed defects, but not within or at the base of these defects. These results suggest that probing of root surfaces may create defects that do not fully remineralize. PMID- 12887150 TI - Smith-Lemli-Opitz syndrome: a review, case report and dental implications. AB - Smith-Lemli-Opitz syndrome (SLOS) is an autosomal recessive developmental disorder characterized by multiple congenital malformations, dysmorphic craniofacial features, and mental impairment. SLOS is caused by a deficiency of the enzyme 7-dehydrocholesterol delta7 reductase which converts 7 dehyrocholesterol to cholesterol. This error results in elevated serum levels of 7-dehydrocholesterol and decreased levels of serum cholesterol. This article describes the clinical features and medical treatment of SLOS. A case report is included, with recommended guidelines for providing safe and comprehensive dental care for individuals with SLOS. PMID- 12887151 TI - Oral manifestations of renal osteodystrophy: case report and review of the literature. AB - Renal osteodystrophy, characterized by uneven bone growth and demineralization, is described. Oral manifestations of the disorder are described, and the value of dental radiographs in early detection of renal osteodystrophy is noted. A case report of a patient with severe oral complications, which resulted from long standing end-stage renal disease and secondary hyperparathyroidism, is presented. Giant cell lesions of hyperparathyroidism, referred to as brown tumors (which may be associated with pain and swelling), are the key clinical oral manifestations and are the most dramatic dental radiographic finding in patients with renal osteodystrophy. Bone changes may include loss of lamina dura, giant cell lesions of hyperparathyroidism, and bone demineralization. The dentist's role in detection, assessment, and treatment is stressed. PMID- 12887152 TI - Effect of soaking prior to cooking on the levels of phytate and tannin of the common bean (Phaseolus vulgaris, L.) and the protein value. AB - The effect of soaking in domestic processing, on the nutritive value of the common bean (Phaseolus vulgaris, L.) cv IAC-Carioca, was studied. Five treatments were carried out with experimental diets, and offered to male, recently weaned Wistar rats. The protein sources were, respectively, control diet (casein) (CC), casein plus the soluble solids found in the soaking water (CSS), freeze dried bean cooked without soaking (BNS), freeze dried bean cooked with the non-absorbed soaking water (BSW), freeze dried bean cooked without the non-absorbed soaking water (BSNW). and an aproteic diet (AP) for corrective purposes. The anti nutritional factors (phytates and tannins), were determined in the differently processed beans and in the soaking water. The following values for the reduction of phytates were obtained: BNS (20.9%), BSNW (60.8%) and BSW (53.0%), and the tannins were reduced by: BNS (86.6%), BSNW (88.7%) and BSW (89.0%). No significant differences were observed between the various treatments using the common bean as protein source, with respect to the net protein ratio (NPR). With respect to the digestibility corrected by non-protein diet, values varying between 94.1% and 94.6% for casein, and between 57.5% and 61.4% for the common bean, were observed, the treatment BNS being more digestible. It was concluded that soaking did not interfere with the NPR of the experimental diets containing the common bean as protein source, nor did it reduce the tannin content. However soaking was capable of reducing the phytate levels in the common bean. On the other hand, soaking was unable to increase the protein digestibility of the common bean, since the treatment BNS showed the highest value for digestibility. PMID- 12887153 TI - Protective effects of mustard leaf (Brassica juncea) against diabetic oxidative stress. AB - Of four fractions (CH2Cl2, EtOAc, BuOH and H2O) from mustard leaf (Brassica juncea), the EtOAc fraction showed the strongest inhibitory effects, which were concentration-dependent, on the formation of advanced glycation end products and free radical-mediated protein damage in an in vitro system, indicating that this fraction has a potential protective role against diabetes and/or its complications. Based on these results, we carried out an in vivo study to determine whether the EtOAc fraction protected against diabetic oxidative stress induced by streptozotocin. Oral administration of the EtOAc fraction at doses of 50 and 200 mg/kg body weight/d for 10 d reduced the serum levels of glucose and glycosylated protein, implying that the impaired glucose metabolism due to diabetes had been ameliorated. In addition, the EtOAc fraction significantly reduced the thiobarbituric acid-reactive substance levels of serum and hepatic and renal mitochondria. Furthermore, the elevated levels of superoxide and nitrite/nitrate were reduced in a dose-dependent manner by oral administration of the EtOAc fraction. These findings suggest that the EtOAc fraction from mustard leaf might be beneficial in attenuating the damage caused by oxidative stress involved in diabetes and its complications. PMID- 12887154 TI - Effect of vitamin E supplementation on antioxidant defense systems and humoral immune responses in young, middle-aged and elderly Korean women. AB - Free radical-mediated oxidative stress has been implicated in the pathogenesis of numerous chronic diseases. Vitamin E is known to play an important role in the free-radical quenching process. However, clinical trials with vitamin E have yielded contrasting results in the prevention of several diseases related to oxidative stress. This study was undertaken to investigate the antioxidative and humoral immunologic effects of vitamin E supplementation in three different age groups: young (mean age 32.7 +/- 5.7 y), middle-aged (mean age 47.0 +/- 5.0 y) and elderly (67.6 +/- 4.7 y) women. Volunteer subjects were given a supplement of 400 IU dl-alpha-tocopherol acetate for 6 wk. Thiobarbituric acid reacting substances (TBARS) in the plasma significantly decreased with vitamin E supplementation. In addition, the radical scavenger activities (RSA) of red blood cells significantly increased with vitamin E supplementation in all age groups. However, humoral immune response modulation was not observed following vitamin E supplementation. Even though there is no clear indication that vitamin E supplementation is necessary to improve the humoral immune functions, vitamin E supplementation may be beneficial to all adult age groups as a preventive measure for complications related to oxidative damage. PMID- 12887155 TI - Effects of feeding oyster, Crassostrea gigas, on serum and liver lipid levels in rats. AB - The effects of feeding dietary and defatted oyster meat on lipid metabolism were investigated in rats by comparing measurements with those of casein and soybean protein. In the first experiment, male rats were fed 0.1% and 1% cholesterol supplemented diets containing casein, oyster or soybean protein under the same dietary level of protein (20%). The concentrations of serum and liver cholesterol in the oyster group were significantly lower than those in the casein group for both the 0.1% and 1% cholesterol-supplemented diets. The cholesterol-lowering effect of oyster meat was more predominant than that of soybean protein. Feeding oyster meat significantly decreased the serum triglyceride concentration as compared to feeding casein for the 0.1% cholesterol-supplemented diets, and it reduced hepatic triglyceride concentration in both groups fed the 0.1% and 1% cholesterol-supplemented diets. The excretion of fecal total steroids was higher in the rats fed oyster meat than those fed casein or soybean protein for both the 0.1% and 1% cholesterol-supplemented diets. In the second experiment, the effects of defatted oyster on lipid metabolism were compared with casein and soybean protein in diets supplemented with cholesterol. The serum cholesterol concentration in the defatted oyster group was comparable to that in the other two groups, but the ratio of high-density lipoprotein-cholesterol to total cholesterol was higher in the defatted oyster group. The feeding of defatted oyster induced a lower liver cholesterol concentration as compared to casein and soybean protein. Serum and liver triglyceride levels were lower in the defatted oyster group than in the casein group. Defatted oyster accelerated the fecal excretion of both neutral and acidic steroids as compared to casein. Our results suggest that the feeding of oysters exerts a more potent hypolipidemic activity than soybean protein, and the effect may be ascribed to both lipid and non-lipid fractions in oyster. PMID- 12887156 TI - Capsaicin increases endurance capacity and spares tissue glycogen through lipolytic function in swimming rats. AB - The influences of various doses of capsaicin on endurance capacity remain to be clarified. The purpose of this study was to determine whether or not capsaicin delays stored tissue glycogen depletion. Rats were orally given either a vehicle or a dose of capsaicin, 6, 10, or 15 mg/kg of body weight, 2 h before exercise. The rats in each group were divided into three subgroups for resting and swimming exercise (30 min, exhaustion). Swimming exercises were performed with a weight corresponding to 3% body weight attached to the tail, and the endurance capacity was evaluated by the swimming time until exhaustion. The 15 mg/kg dose of capsaicin significantly (p < 0.05) enhanced the endurance performance time and plasma concentration of epinephrine, norepinephrine, free fatty acid and glucose rose to significantly higher levels within 30 min; swimming exercise compared to rest (p < 0.05). At the 15 mg/kg capsaicin dosage. the plasma insulin level decreased to significantly lower levels in group subjected to 30-min swimming as compared to the resting group (p < 0.05), while plasma glucagon rose to a significantly higher level (p < 0.05). Liver and gastrocnemius muscle glycogen in the group subjected to 30-min swimming was maintained at significantly higher concentrations in the rats fed 15 mg/kg of capsaicin as compared to the vehicle counterparts (p < 0.05). These results suggest that the improvement in swimming endurance with the high capsaicin dosage is caused by an increase in fatty acid utilization as the energy source, resulting in the sparing of glycogen. PMID- 12887157 TI - Further evidence regarding the effect of dietary protein on oral tolerance against beta-lactoglobulin through Th1-mediated immune response in mice. AB - Oral tolerance is a potential strategy for preventing or minimizing aberrant immune responses. Although, oral tolerance has been extensively studied, to date the effects of dietary protein on the induction of oral tolerance are poorly understood. We have previously shown that restricted dietary protein induces oral tolerance to ovalbumin. This study was designed to investigate whether or not such tolerance occurs with beta-lactoglobulin (BLG) instead of ovalbumin (OVA) and if the tolerance resulting from this feeding regimen involves Th1-mediated immune response. Female BALB/c mice fed either 20% or 5% dietary protein were given 5 mg BLG or water orally for four consecutive days and then immunized intraperitoneally (ip) twice with BLG at 3-wk intervals. Oral tolerance induction was compared in BLG-fed and water-fed mice by measuring total IgE, BLG-specific antibodies, footpad reactions, splenocyte proliferation, and cytokine production. When mice were given BLG orally before ip immunization, the Th1-mediated immune responses (production of IL-2, IFN-gamma, and IgG2a) were significantly reduced, whereas the Th2-mediated immune responses (production of IL-4 and IgG1) were unchanged. The Th1-mediated immune responses were markedly down-regulated in mice fed 5% protein as compared to those in mice fed 20% protein. Moreover, the production of total IgE, BLG-specific IgE, splenocyte proliferation, and footpad reactions were more reduced in mice fed 5% protein than those in mice fed 20% protein. The present study provides evidence that dietary protein plays an important role in the induction of oral tolerance against BLG as the result of, clear down-regulation of Th1 helper activity accompanied by a reduction in IgE. PMID- 12887158 TI - The role of exercise physical activity in varying the total energy expenditure in healthy Japanese men 30 to 69 years of age. AB - This study was designed to examine 1) the role of exercise physical activity (EPA), and then 2) physical fitness and body composition upon variation of the total energy expenditure (TEE) in healthy Japanese men aged 30 to 69 y (n = 40). EPA and TEE were assessed over 14 d using an accelerometer and a doubly labeled water (DLW) method, respectively. Basal metabolic rate (BMR) was measured after 10 h fasting on the morning of the day of DLW dosing. Physical activity-induced energy expenditure (PAEE) was calculated by subtracting BMR and diet-induced thermogenesis (DIT = 101 TEE) from TEE. EPA was subdivided into three intensities: low, moderate and high and the accumulated duration (time expressed in minutes) of each of these was calculated. Body composition and physical fitness (VO2max) were determined using an underwater weighing method and a treadmill exercise test, respectively. BMR (mean +/- SD: 1,459 +/- 181 kcal/d) declined significantly with age (r = -0.37. p < 0.05), but PAEE (946 +/- 320 kcal/d) and TEE (2,672 +/- 369 kcal/d) did not. A multiple stepwise regression analysis was used to develop an empirical model that relates energy expenditure measured by the DLW (TEE) to age, height, body mass index, FM, FFM, percentage body fat, VO2max, and accumulated duration spent for low-, moderate-, and high intensity EPA. The results revealed that FFM and high-intensity EPA were identified as important determinants of TEE and accounted for 51%. We may therefore conclude that 1) high-intensity EPA appears to be relevant in determining TEE, especially among active individuals, and 2) body composition was more important than physical fitness in determining TEE in this population. PMID- 12887159 TI - Analysis of the comprehensive effects of polyunsaturated fatty acid on mRNA expression using a gene chip. AB - To investigate the comprehensive effects of polyunsaturated fatty acids (PUFA) on gene expression, we analyzed changes of mRNA expression in PUFA-treated HepG2 cells using a DNA micro array. We incubated HepG2 cells for 24 h with or without 0.25 mM oleic acid (OA), arachidonic acid (AA), eicosapentaenoic acid (EPA) or docosahexaenoic acid (DHA), and then compared the expression profiles of thousands of genes using a GeneChip. PUFA influenced the expression of various genes related to cell proliferation, growth and adhesion, as well as for many transcription factors including sterol regulatory element binding proteins (SREBP). Treatments with AA, EPA, and DHA repressed the expression of genes related to cholesterol and lipid metabolism. Moreover, data from gene chip analysis proved that PUPA reduced the expression ofprostasin, which is a serine protease. By measuring the mRNA levels of SREBPs, mevalonate pyrophosphatase and prostasin using quantitative RT-PCR, we confirmed the effect of PUFA revealed by gene chip analysis. These data might provide useful clues with which to explore novel functions of PUPA. PMID- 12887160 TI - Dietary patterns and further survival in Japanese centenarians. AB - We have previously reported that centenarians (persons > or = 100 y old) in Tokyo prefer dairy products. Dietary preferences may be associated with longevity. The aim of the present study was to investigate the relationship between dietary patterns and further survival in centenarians. During 1992-1999, we examined the dietary practices of 104 centenarians (29 men and 75 women; mean age, 100.3 +/- 0.9 y) who lived in the Tokyo metropolitan area. Dietary patterns were classified by kappa-means cluster analysis. As clinical co-variables, we considered activities of daily living, cognitive function, nutritional status, presence of important disease, gender, and age at the time of the initial survey. Survival data were recorded yearly until 2001, and then tested with Kaplan-Meier analysis and the log rank statistic. Four dietary patterns were identified: a pattern preferring vegetables (n = 33), a pattern preferring dairy products (n = 26), a pattern preferring beverages (n = 10), and a pattern preferring cereals (n = 35). No clinical variables differed between the four dietary patterns. In 2001, 28 centenarians were still alive. The survival rate for those preferring dairy products was the highest of the four dietary patterns; in particular, being significantly higher than the pattern preferring beverages (p = 0.048). A dietary pattern preferring dairy products was associated with increased survival in Tokyo area centenarians. PMID- 12887161 TI - Dietary diacylglycerol-rich oil stimulation of glucose intolerance in genetically obese rats. AB - The effects of dietary 1,3-diacylglycerol-rich oil (DG oil) on glucose and lipid metabolism were investigated in comparison with triacylglycerol (TG) oil in female genetically obese Wistar fatty rats. The obese rats and their lean littermates (8 wk old) were fed a synthetic diet containing 10%, (w/w) DG or TG oil for 5 wk. The body weights, abdominal fat weights, and the plasma and liver TG concentrations were not significantly different due to dietary fat type in the obese and lean rats. The plasma glucose concentrations were significantly elevated by dietary DG oil as compared to TG oil in the portal vein and inferior vena cava of obese and lean rats. The plasma free fatty acid concentrations were markedly elevated by dietary DG oil as compared to TG oil in the portal vein and inferior vena cava of both genotype rats, particularly in the obese rats. In the glucose tolerance test, the obese rats fed DG oil showed glucose intolerance, possibly due to the markedly elevated plasma free fatty acids. Thus, the effects of dietary DG oil on lipid-lowering effects and anti-obesity were not observed in either genotype in the present study. Moreover, it is remarkable that glucose intolerance was induced by dietary DG oil in the genetically obese rats. dietary PMID- 12887162 TI - Studies on the effect of monosodium glutamate [MSG] administration on some antioxidant enzymes in the arterial tissue of adult male mice. AB - The subcutaneous administration of monosodium glutamate to normal adult male mice at dose levels of 4 and 8 mg/g body weight caused a significant increase in lipid peroxidation level in the arterial tissue. The levels of total sulfhydryl and protein-bound sulfhydryl groups were found to be significantly increased, whereas non-protein-bound sulfhydryl, representing the glutathione level, was significantly decreased. It was also observed that the administration of monosodium glutamate at a dose level of 4 mg/g body weight and above induced oxidative stress by significantly lowering the activities of antioxidant enzymes like superoxide dismutases, catalase, and glutathione metabolizing enzymes like glutathione reductase and glutathione peroxidase in the arterial tissue. PMID- 12887163 TI - Prevention of type 2 diabetes. AB - The prevalence of diabetes is increasing worldwide. The World Health Organisation has estimated that there will be around 300 million diabetics by 2025. The largest increase will occur in Asia. The prevalence of type 2 diabetes is increasing due to a combination of factors: increasing lifespan, sedentary lifestyle, excessive intake of high energy foods, increasing prevalence of overweight/obese people. The Finnish Diabetes Prevention Study Group has clearly shown that changes in the lifestyle of both overweight men and women with impaired glucose tolerance can reduce the incidence of type 2 diabetes by 58%. This finding was confirmed by the Diabetes Prevention Programme which found that lifestyle intervention in individuals with impaired fasting glucose or impaired glucose tolerance reduced the risk of developing type 2 diabetes by 58%, whereas treatment with metformin reduced the risk of type 2 diabetes by only 31%. Both acarbose and troglitazone have also been shown to reduce the progression to diabetes in individuals who are at high risk of developing type 2 diabetes. Since the cure for diabetes remains some way off our concerted efforts should be directed at prevention of diabetes in order to curb the increasing prevalence of diabetes worldwide. Lifestyle changes are more beneficial than long term drug therapy in the prevention of diabetes and should be actively promoted. PMID- 12887164 TI - Tocotrienols-rich diet decreases advanced glycosylation end-products in non diabetic rats and improves glycemic control in streptozotocin-induced diabetic rats. AB - This study determined the effects of palm vitamin E (TRF) diet on the levels of blood glucose, glycated hemoglobin (gHb), serum advanced glycosylation end products (AGE) and malondialdehyde (MDA) of diabetic Sprague-Dawley rats. The rats received either control (normal rat chow), TRF diet (normal chow fortified with TRF at 1 g/kg) or Vitamin C diet (vitamin E-deficient but contained vitamin C at 45 g/kg). The animals were maintained on the respective diet for 4 weeks, made diabetic with streptozotocin (STZ), then followed-up for a further 8 weeks. At week-4, mean serum AGE levels of rats given TRF diet (0.7 +/- 0.3 units/ml) were significantly lower than those of control or Vitamin C diet rats (p pounds 0.03). The levels increased after STZ and became comparable to the other groups. At week 12, blood glucose (20.9 +/- 6.9 mM) and gHb (10.0 +/- 1.6%) of rats on TRF diet remained significantly low compared to that of control or Vitamin C diet rats (p pounds 0.03). MDA however, was not affected and remained comparable between groups throughout the study. This study showed that TRF may be a useful antioxidant; effectively prevented increase in AGE in normal rats, and caused decrease in blood glucose and gHb in diabetic rats. Further studies are needed to elucidate the mechanisms of action of TRF. PMID- 12887165 TI - Risk assessment and microbiological profile of infections in paediatric cancer patients with febrile neutropenia. AB - Febrile neutropenia is a common and potentially fatal problem encountered in cancer patients undergoing chemotherapy. We carried out an observational study to evaluate the possible risk factors of developing fever amongst neutropenic children with an underlying malignancy. We also looked at the microbiological profile of causative pathogens in patients with febrile neutropenia. During a study period of 1 year, a total of 90 neutropenic episodes were recorded amongst 57 patients who were on treatment and follow-up during the study period. Multivariate analysis showed that factors such as chemotherapy status, underlying disease, existing central venous catheters, presenting white blood cell counts at chemotherapy, use of steroid therapy or hospitalisation at the onset of neutropenia, were not significant risk factors for developing fever during neutropenic episodes. Although the presence of a central venous catheter was associated with a higher risk of developing fever, it did not reach statistical significance (p=0.11). Of the 90 neutropenic episodes, 59 (65.6%) developed fever and 25 of these had positive blood cultures. The causative organisms include gram negative bacteria (64%), gram positive bacteria (16%) and fungus (20%). Of the gram-negative organisms, Klebsiella spp. predominated (28%) with the extended spectrum beta-lactamase producing strain forming the majority (16%). Amongst those with fungaemia, Candida spp. and Candida tropicalis formed the majority (8% each) of the isolates. PMID- 12887166 TI - Infrequent occurrence of vancomycin-resistant enterococci in poultry from Malaysian wet markets. AB - Fifty samples of chicken, duck and geese faeces were obtained from 13 wet markets in Kuala Lumpur to study the prevalence of vancomycin-resistant enterococci (VRE) among local market poultry. Biotyping of colonies grown on azide agar incubated at 45 degrees C yielded E. pseudoavium, E. faecalis, E. faecium and E. gallinarum from chicken faeces and E. malodoratus, E. faecalis, E. faecium, E. gallinarum, E. hirae/dispar, and E. durans from goose and duck faeces. On agar containing 6 mg/ l of vancomycin, one strain of E. flavescens was identified, giving a VRE detection rate of 2.0%. This isolate had a vancomycin M.I.C. of 8 mg/l as determined by the Etest, and the van C-3 gene that was identified by PCR followed by sequence analysis. The prevalence of VRE among poultry sold in local markets appears to be low, and may reflect the infrequent use of antimicrobials in our poultry farms. Nevertheless, the possibility of human acquisition of microbes via the food chain cautions against the use of antimicrobials in animal husbandry that may encourage the emergence and spread of multi-drug resistant organisms like the VRE among animal microbial flora. PMID- 12887167 TI - The prevalence of GP Mur and anti-"Mia" in a tertiary hospital in Peninsula Malaysia. AB - The Mi III phenotype of the Miltenberger subsystem (or GP Mur) is relatively common in Southeast Asia especially along the south-east coast lines of China and Taiwan. The term anti-"Mia" describes antibodies that react with the Mi III phenotype. Since the Peninsula Malaysian population is a multiethnic one with a significant proportion of Chinese, a study was conducted into the prevalence of anti-"Mia" in patients from its 3 major ethnic groups--Chinese, Malays and Indians, as well as the GP Mur phenotype in blood donors (healthy individuals). Blood samples from 33,716 patients (general and antenatal) were screened for anti "Mia" from January 1999 to December 2000. The investigation for the GP Mur phenotype representing the corresponding sensitizing antigen complex was carried out in 655 blood donors. Serum anti-"Mia" antibody was found to be the third most commonly occurring antibody detected in our patients and was found in all the ethnic groups. The antibody was detected in 0.2% of 33,716 antenatal and general patients with a prevalence in Chinese of 0.3%, Malay 0.2% and Indian 0.2%. The detection of these antibodies in the ethnic groups other than the Chinese is a noteworthy finding as such information is not well documented. The GP Mur red cell phenotype was detected in 15/306 (4.9%) of Chinese blood donors, a lower prevalence than in Chinese populations in other countries in the region. More significant was its detection in the Malays (2.8%) and the Indians (3.0%). Because of the many reports of clinical problems associated with the "Mia" antibody including the causation of fetal hydrops and haemolytic transfusion reactions, it is warranted that the GP Mur red cells be included in screening panels for group and screen procedures in countries with a significant Asian population. PMID- 12887168 TI - Anaemia and iron status among blood donors in a blood transfusion unit in Malaysia. AB - Iron deficiency is a major complication of regular blood donation as a result of regular iron loss from each donated blood unit. Ninety-two regular blood donors and 95 first time blood donors attending a hospital-based blood transfusion centre were assessed as to their haematological and iron status by blood counts and serum ferritin levels as an indicator of iron stores. All donors had passed the haemoglobin-screening test using a copper sulphate method prior to blood donation. Ferritin levels were found to be significantly lower among regular blood donors (47.8 mmol/L) as compared to first time blood donors (94.2 mmol/L). Iron deficiency as observed by low ferritin levels was seen in 7.4% of all first time donors as compared to 17.4% in regular donors. Male first time donors showed a low prevalence of iron deficiency but the prevalence significantly increased with regular blood donation. Female first time and regular blood donors however did not show any significant differences in prevalence of iron deficiency, with both groups exhibiting prevalence rates similar to male regular donors. The association between haemoglobin levels and iron deficiency was poor and the copper sulphate-screening test was found insensitive to anaemia with many donors passing the test and donating blood despite being anaemic. It is concluded that a high prevalence of iron deficiency is present among regular male blood donors and all female donors. Besides, the use of the copper sulphate screening test as a sole criterion for anaemia screening should be reviewed. Ferritin measurements should be included in the routine assessment of blood donors especially among regular blood donors. PMID- 12887169 TI - Isolation of Neissseria meningitidis from an unusual site. AB - A 59-year-old post-menopausal lady who had returned from a pilgrimage to Mecca about a month earlier presented with a three days' history of profuse vaginal discharge. Neissseria meningitidis was isolated from high vaginal swab specimens taken from her on 2 occasions, five days apart. Her symptoms disappeared without treatment after two weeks. We conclude that although the organism may have been a colonizer, it is possible that it was responsible for the self-limiting genital infection in this patient. PMID- 12887170 TI - Papillary cystic type of acinic cell carcinoma of parotid: fine needle aspiration cytological features of a high grade variant with oncocytic metaplasia. AB - A 60-year-old female developed a right parotid swelling six months after surgery for intra-oral squamous cell carcinoma. Fine needle aspiration (FNA) cytological smears showed dissociated large and small pleomorphic tumour cells with abundant mitoses and oncocytic features. A cytological diagnosis of parotid acinic cell carcinoma (ACC) was made. Histological study of the subtotal parotidectomy specimen showed a papillary cystic variety of acinic cell carcinoma (ACC-PCV). FNA cytological features in this case of ACC-PCV differs from the two previously reported cases in that it showed prominent oncocytic and high grade features and absence of papillary pattern in the cytological smears. ACC-PCV is an uncommon tumour and knowledge of its varied FNA cytological features is important for the diagnosis of this neoplasm. PMID- 12887171 TI - Calibre persistent submucosal artery of the jejunum: a rare cause of massive gastrointestinal bleeding. AB - Historically a calibre persistent submucosal artery was most often described in the stomach. However in later years it was also discovered in the duodenum and jejunum. It is an uncommon and important cause of massive gastrointestinal bleeding in which failure of detection and early intervention would lead to death. In this paper we report a 27-year-old man with no significant medical history who presented at the emergency unit for recurrent melaena, haematochezia and hypotension. Initial investigations failed to localize the source of bleeding. Emergency exploratory laporatomy revealed a small jejunal mucosal nodule that was actively spurting blood. Histopathological evaluation identified a calibre persistent submucosal artery. PMID- 12887172 TI - Current concepts in the treatment of acne: report from a clinical roundtable. AB - Acne vulgaris is the most common skin disorder in the United States, affecting approximately 17 million people. This figure includes 80% of all persons between the ages of 11 and 30 years, regardless of race, ethnicity, or gender (Kraning and Odland, 1979). Acne has a significant economic and social impact on doctor visits, medications, and absenteeism, as well as in the negative effect it can have on self-image and outlook, especially during the emotionally critical period of adolescence. Persons with acne may think of themselves as unworthy and socially unacceptable. Severe acne may lead to scarring and disfigurement, aggravating the already present psychosocial aspects of this condition. Because of the multifactorial nature of acne, current treatment options aim to modify one or more of its pathogenic factors. Clinical experience has shown that combination therapies, which affect multiple aspects of pathophysiology, are most likely to achieve this goal and help in its long-term management and resolution. PMID- 12887173 TI - New therapy update--A unique combination formulation in the treatment of inflammatory acne. AB - Duac Topical Gel (clindamycin 1%-benzoyl peroxide 5%) is a uniform, aqueous gel manufactured under the US Food and Drug Administration (FDA) Current Good Manufacturing Practice (cGMP) regulations and controls, which ensure that consistent product is delivered to the patient every time. Since the product is premixed, the patient does not have to wait for mixing or compounding by the pharmacist. The active ingredients have been shown to be stable for 60 days at room temperature, so that neither the patient nor the physician needs to store the product in the refrigerator. When kept at temperatures of 2 degrees C to 8 degrees C, the product is stable for 2 years. In a clinical study of 358 subjects with moderate to moderately severe acne vulgaris, once-daily treatment with clindamycin 1%-benzoyl peroxide 5% for 11 weeks reduced inflammatory lesions by 53% and noninflammatory lesions by 25%. Good or excellent global response was experienced in 50% of subjects. Overall tolerance ratings were good to excellent in 99% of subjects, and, except for mild to moderate expected local reactions, there were no adverse events related to treatment. PMID- 12887174 TI - A PCR-RFLP test to detect allelic variants of the bovine kappa-casein gene. AB - Point mutations in exon IV of bovine kappa-casein gene (kappaCn, CASK, CSN3) determine nine allelic variants (A, B, C, E, F, G, H, I, and A1) for the gene. These variants are associated with major differences in composition and manufacturing properties of milk (i.e., cheese yield). A PCR-RFLP test was developed in order to distinguish the different alleles. Polymorphisms are detected by digestion with the endonucleases HindIII, HaeIII, and MaeII followed by electrophoresis in agarose gels stained with ethidium bromide. Twenty eight DNA samples from different breeds of Argentina were analyzed for the A, B, and E variants. This simple PCR-RFLP test makes feasible the inclusion of kappa-casein genotypes in breeding plans. PMID- 12887175 TI - Molecular characterization of lysozyme type II gene in rainbow trout (Oncorhynchus mykiss): evidence of gene duplication. AB - Rainbow trout (Oncorhynchus mykiss) have two types of lysozyme. Type II lysozyme differs from type I by only one amino acid, but only type II lysozyme has significant bactericidal activity. Due to this novel antibacterial property, lysozyme type II appears to be a candidate gene for enhancing disease resistance in fish as well as livestock species. Using polymerase chain reaction the lysozyme type II gene was amplified from genomic DNA isolated from rainbow trout. Two amplified fragments of 2041 and 2589 bp were observed. Sequencing revealed both amplicons were lysozyme genes having nearly identical nucleotide sequences, except the longer fragment has 548 base pairs inserted in intron 2 at nucleotide position 513 and a few point mutations within intron 2. Both versions of trout lysozyme type II gene were comprised of four exons and three introns. We also demonstrated that trout lysozyme is most likely encoded by these two different genes. PMID- 12887176 TI - Porcine 11beta-hydroxysteroid dehydrogenase type 2 isoform: complete coding sequence and polymorphisms. AB - 11Beta-hydroxysteroid dehydrogenase type 2 (11beta-HSD2) is involved in the regulation of the peripheral glucocorticoid concentrations. Due to the central role of glucocorticoids in protein turnover, 11beta-HSD2 is a candidate gene for optimising production traits in livestock. In addition, mutant 11beta-HSD2 animals may be used as models for human disorders. Here, we present the complete porcine 11beta-HSD2 coding sequence, the RT-PCR strategy for the examination of the coding sequence and the polymorphisms found in the pig. PMID- 12887177 TI - Cloning and expression analysis of chicken phospholipid-hydroperoxide glutathione peroxidase. AB - Phospholipid-hydroperoxide glutathione peroxidase (GPX4 or PHGPX) is a unique selenium dependent glutathione peroxidase that reduces phospholipid, cholesterol, and cholesteryl ester hydroperoxides. Phospholipid-hydroperoxide glutathione peroxidase has been shown to exist as both a 197 amino acid mitochondrial targeting protein and as a 170 amino acid non-mitochondrial protein. The cDNA encoding the non-mitochondrial chicken GPX4 (cGPX4) has been isolated from an immortalized DF-1 chicken embryonic fibroblast (CEF) cell line cDNA library. The nucleotide sequence of cGPX4 was 802 bp in length with an open reading frame (ORF) that encoded 170 amino acids but lacked the N-terminal domain that encoded the mitochondrial leader sequence (MLS). Chicken non-mitochondrial GPX4 was highly expressed in brain and stromal tissues. Surprisingly, it was found that ovarian stromal tissue cGPX4 expression is regulated quite differently according to the reproductive status of the bird, suggesting that GPX4 may play an important role in reproduction in response to steroid hormones, in addition to its general antioxidant functions. PMID- 12887178 TI - Patterns of cellular gene expression in cells infected with cytopathic or non cytopathic bovine viral diarrhea virus. AB - Bovine viral diarrhea virus (BVDV) infection in cattle is responsible for mucosal disease; an invariably fatal syndrome characterized by the recovery of two BVDV strains: cytopathic (cp) or noncytopathic (ncp). To understand the cellular responses to cp BVDV infection, we carried out differential display-polymerase chain reaction (DD-PCR) analysis of gene expression in infected cells. Altered expression of 14 genes involved in several functions was observed in cells infected with cp BVDV: (1) immune regulation, such as CD46, FKBP-12, and osteopontin (OPN); (2) apoptosis-related cysteine proteases like calpain; (3) signaling plasma membrane proteins such as integrin beta1, and prion protein; and (4) unknown function genes. Northern blot analysis of the expression of these genes in ncp BVDV infected cells revealed that while the expression of some genes was affected as in cp BVDV infected cells, others show a clearly contrary change. We postulate that a cause-effect relationship may exist between the differential gene expression alterations that characterize cp and ncp BVDV infections and the unique diseases associated with each BVDV biotype. PMID- 12887179 TI - Construction of YAC/BAC contig map for the BTA 6q21 region containing a locus for bovine chondrodysplastic dwarfism. AB - To characterize the bovine chromosome 6q21 for bovine chondrodysplastic dwarfism (BCD), we developed 48 new microsatellite markers from yeast artificial chromosome (YAC) and bacterial artificial chromosome (BAC) clones using a modified magnetic bead capture method. These new markers were used to construct a high-resolution physical map of the region with a total of 85 loci. The physical map will be a powerful tool for successful positional cloning experiments. PMID- 12887180 TI - Candidate gene approach: potentional association of caspase-1, inhibitor of apoptosis protein-1, and prosaposin gene polymorphisms with response to Salmonella enteritidis challenge or vaccination in young chicks. AB - Salmonella enteritidis (SE) contamination of poultry products is a major cause of foodborne disease worldwide. Caspase-1 and inhibitor of apoptosis protein-1 (IAP 1) were selected as candidate genes for chicken response to SE because their proteins play critical roles in the apoptotic pathway when intracellular bacteria interact with host cells. Prosaposin (PSAP) was selected as a positional candidate gene based on a previous quantitative trait loci (QTL) linkage study using the same population. The F1 offspring of outbred sires crossed with three diverse, highly inbred dam lines (two major histocompatibility complex-congenic Leghorn lines named G-B1 and G-B2, and one Fayoumi line) were used to define the phenotypes. The F1 birds were involved in either pathogenic SE challenge, in which spleen and cecum content bacterial load were quantified, or SE vaccination, in which plasma antibody level to SE vaccine was evaluated. A polymerase chain reaction-restriction fragment length polymorphisms (PCR-RFLP) assay was developed to identify single-nucleotide polymorphism (SNP) in the three genes. The F1 offspring of heterozygous sires for each gene were genotyped. The sire caspase-1 gene was significantly associated with cecum content bacterial load (P = 0.04) in the three combined dam line crosses, and with spleen bacterial load in the G-B1 cross (P=0.02). The sire caspase-1 gene was also significantly associated with antibody level to SE vaccine (P=0.03) in F1 males in the three combined dam line crosses. The sire IAP-1 gene was significantly associated with spleen bacterial load (P=0.04) in the three combined dam-line crosses, and interacted with dam line genetics (P = 0.01) for cecum content bacterial load. The sire PSAP gene significantly interacted with sex for spleen bacterial load (P = 0.004). This study is the first to demonstrate the association of SNPs for caspase-1, IAP-1, and PSAP genes with SE vaccine and with pathogen challenge response in chickens. PMID- 12887181 TI - Allerton III. Beyond livestock genomics. AB - Throughout the Allerton III Conference, several consistent research needs were identified across scientific disciplines. First, additional basic research is needed to identify genomic mechanisms and novel genes/proteins in a variety of tissues under different conditions. Second, expansion of the infrastructure of the scientific community is needed. This can best be accomplished by additional competitive grants programs for training grants, program project grants, and multidisciplinary research projects. Third, the need for improved tools for animal bioinformatics was emphasized. Fourth, competitive grants programs for extension/outreach efforts and application of genomic technologies to production systems are needed. Finally, efforts to publicize and document the benefits of animal genomics for improved human health and animal production systems to members of Congress and the general public should be enhanced. PMID- 12887182 TI - Chromosomal mapping of HSPCB and MYL1 expressed abundantly in the bovine fetus. AB - Chromosomal mapping of expressed sequence tags for HSPCB and MYL1 expressed abundantly in the bovine fetus was performed by analyzing bovine/murine somatic cell hybrid DNAs with polymerase chain reaction (PCR) using primers specific for those 3'-untranslated regions. HSPCB and MYL1 were assigned to bovine chromosomes 23 and 2, respectively. PMID- 12887183 TI - Mapping of porcine genetic markers generated by representational difference analysis. AB - Representational difference analysis (RDA) was performed using pig genomic DNA from a Landrace non-selected control population and a Landrace population selected for increased loin muscle area (LMA) for five generations. Pigs used for the analysis differed phenotypically for various carcass traits and were divergent in genotype at the skeletal muscle ryanodine receptor 1 locus. Two RDA experiments were performed using BamHI and BglII. Fourteen BamHI and 37 BglII difference products were cloned and sequenced. Oligonucleotide primers were designed to amplify RDA difference products and sequence-tagged sites (STS) were developed for 16 RDA fragments (two BamHI and 14 BglII). These 16 STS were mapped using the INRA-Minnesota porcine Radiation Hybrid panel. Polymorphisms identified in nine of the STS were used to place these markers on the PiGMaP genetic linkage map. Sequence-tagged sites were localized to 11 different chromosomes including three markers on chromosome 11 and four markers on chromosome 14. Development of RDA markers increases the resolution of the pig genome maps and markers located within putative quantitative trait locus (QTL) regions can be used to refine QTL positions. PMID- 12887184 TI - Reducing airborne pathogens and dust in commercial hatching cabinets with an electrostatic space charge system. AB - Commercial hatcheries typically infuse hydrogen peroxide or formaldehyde gas into hatching cabinets to reduce airborne pathogens that may lead to disease transmission during the hatch. A nonchemical option, an electrostatic space charge system (ESCS), was customized for full-sized commercial hatching cabinets and was tested extensively in broiler hatcheries. The ESCS cleans air by transferring a strong negative electrostatic charge to dust and microorganisms that are aerosolized during the hatch and collecting the charged particles on grounded plates or surfaces. In studies with three poultry companies, the ESCS resulted in significant (P < 0.0001) reductions of airborne dust of 77%-79%, in Enterobacteriaceae and fungus levels not significantly different (P > or = 0.05) from those with formaldehyde, and in 93%-96% lower Enterobacteriaceae than with no treatment or with hydrogen peroxide treatment (P < 0.01). The ESCS significantly (P < 0.05) reduced airborne Salmonella by 33%-83% compared with no treatment or hydrogen peroxide treatment. Results of this study suggest that the ESCS is a viable alternative to chemical treatment for reducing airborne pathogens in full-sized commercial hatchers, and it also provides dust control and containment, which should be helpful in reducing cross contamination and loading of ventilation ducts within different areas of the hatchery. PMID- 12887185 TI - Experimental infection of white-leghorn cockerels with Macrorhabdos ornithogaster (Megabacterium). AB - Macrorhabdos ornithogaster is a newly described anamorphic ascomycetous yeast that has been reported to cause a chronic, debilitating disease in many species of birds, including poultry. Study of this organism is complicated by the limited ability to grow M. ornithogaster in vitro. In this study, we showed that the chicken can be used to amplify this organism and as a model to study its pathogenicity. An infection rate of 100% was achieved in day-old chicks orally inoculated with 10(5) M. ornithogaster derived from the budgerigar (Melopsittacus undulatus). The organism was also determined to increase in number by greater than 10-fold 14 days after oral inoculation in these chicks. Chickens infected with M. ornithogaster demonstrated no sign of illness but had decreased feed conversion efficiency and consistent and characteristic histopathologic lesions in the proventriculus and isthmus of the stomach, suggesting that M. ornithogaster may represent a potential threat to the poultry industry. PMID- 12887186 TI - Efficacy of live virus vaccines against infectious laryngotracheitis assessed by polymerase chain reaction-restriction fragment length polymorphism. AB - The efficacy of four different commercial live vaccines (vaccines A, B, C, and D) against the infectious laryngotracheitis virus (ILTV) was assessed in specific pathogen-free (SPF) chickens. SPF chickens were vaccinated intraocularly at 6 wk old with ILTV live vaccines and were challenged intratracheally with the N91B01 strain of virulent Korean ILTV 2 wk after vaccination. The immunity against ILTV live vaccines was assessed by the incidence of latent infection by the challenge virus in the chickens' tracheas and trigeminal ganglia, the reisolation rate of the challenge virus, and the clinical signs in the chickens challenged with the N91B01 strain of ILTV. The latent infection in chickens was assessed by nested polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Our data showed that the clinical signs and challenge virus isolation were negative in all chickens receiving four difference commercial ILTV live vaccines. The viral DNA of the vaccine strain, but not that of the challenge virus, was detected in chickens vaccinated with vaccine A by nested PCR-RFLP. The viral DNAs of both the vaccine and challenge strains were detected from chickens vaccinated with vaccines B, C, and D. This study showed that only vaccine A can protect chickens from latent infection with the field virulent ILTV. We speculate that the efficacy of infectious laryngotracheitis live vaccines to protect chickens from latent infection with virulent ILTVs can be assessed by nested PCR-RFLP analysis. PMID- 12887187 TI - Protection of chickens from infectious bronchitis by in ovo and intramuscular vaccination with a DNA vaccine expressing the S1 glycoprotein. AB - We have constructed a DNA vaccine (pDKArkS1-DPI) expressing the S1 glycoprotein (Arkansas DPI) of infectious bronchitis virus (IBV) to examine protective immunity after in ovo and intramuscular DNA immunization. Birds receiving in ovo DNA followed by live virus vaccination at 2 wk of age were 100% protected from clinical disease. Birds receiving only live virus vaccine or only in ovo DNA vaccination were < or = 80% protected. IBV was detected up to 10 days postchallenge in unvaccinated control groups, whereas birds receiving in ovo DNA and live virus vaccination cleared IBV from tracheal samples before day 5 postchallenge. Transcription of the S1 gene was confirmed in lung tissue after in ovo vaccination by an antisense riboprobe, and the S1 protein was detected by immunohistology in the heart and bursa. In a separate experiment, birds were injected intramuscularly with either 50, 100, or 150 microg of the DNA vaccine at 1 day of age and then again with either 100, 200, or 300 microg of the DNA vaccine, respectively, at 14 days of age. At 10 days postchallenge, no clinical signs were observed and no challenge virus was reisolated from the birds vaccinated with 150 microg and 300 microg of DNA. Between DNA-vaccinated birds and nonvaccinated control birds, no statistical differences were observed for IBV specific serum antibodies as detected by enzyme-linked immunosorbent assay or the virus neutralization test. These data indicate that DNA vaccination with the S1 gene either in ovo or intramuscularly can provide birds with some protection against clinical disease after homologous IBV challenge. PMID- 12887188 TI - A consideration of previously uncharacterized fowl poxvirus unidirectional and bidirectional late promoters for inclusion in homologous recombinant vaccines. AB - Because of the limited analysis of fowl poxvirus (FPV) promoters, expression of foreign proteins by recombinant FPV has usually been directed by heterologous vaccinia virus or synthetic poxvirus promoters. Thus, the impact of completely homologous recombinant virus vaccines has yet to be realized by the poultry industry. In an effort to increase the availability of such transcriptional regulatory elements, the modulation of gene expression by six previously uncharacterized FPV late promoters was examined. To simplify this comparison, each promoter region was separately coupled to the same reporter gene (lacZ) in individual plasmid constructs, and their activities in transfected, virus infected cells were monitored. In each of the four selected unidirectional transcriptional regulatory elements as well as a 30-base pair representative of the bidirectional promoter region, the predicted temporal specificity of expressing at late stages of virus replicative cycle was verified. Stable lacZ gene transcripts arising from each plasmid varied less than threefold in quantity, whereas the amounts of beta-galactosidase product ranged within a 130 fold interval. Only the promoter that naturally regulates expression of the A type inclusion body protein gene directed production of beta-galactosidase at a level comparable with that associated with the strong vaccinia virus P11 promoter. Because one of the remaining unidirectional transcriptional regulatory elements, P174, was only 2.4-fold less efficient, both of these promoters, P174 and P190, should be satisfactory for directing the expression of poultry pathogen genes inserted into the genomes of FPV recombinant vaccines. PMID- 12887189 TI - Persistence of chicken herpesvirus and retroviral chimeric molecules upon in vivo passage. AB - Mareks disease virus (MDV), a herpesvirus, and avian leucosis virus subgroup J (ALV-J), a retrovirus, were used for experimental coinfection of chickens. Chimeric molecules having sequences of both viruses were detected by the hotspot combined polymerase chain reaction (HS-cPCR) system. The detection of chimeric molecules provided evidence for avian retroviral inserts in the herpesvirus genome. The persistence of chimeric molecules on in vivo passage served to indicate the infectivity of the recombinant virus. The evaluation of formation and persistence of the chimeric molecules was performed in two trials involving three in vivo passages. The chimeric molecules were identified according to the primer sets, their product length, and pattern. The persistence of chimeric molecules on in vivo passages served as an indication of their ability to replicate in and infect chickens. In the first experimental passage, MDV and ALV J prototype strains, MD11 and HC-1, were intraperitoneally (i.p.) injected into 1 day-old chicks. The second trial included two passages. Passage II chicks were injected i.p. and passage III chickens were in contact with the chickens of passage II. For passage II, enriched white blood cells from blood samples of chickens from the first trial that had chimeric molecules were injected i.p. into 1-day-old chicks. For passage III, uninfected chicks were included together with the infected chicks. Synthesis evidence for the various species of chimeric molecules was assessed in the tissues of birds of the second trial. DNA was extracted from blood and feathers and analyzed by the hotspot-combined PCR and by pulsed field gel electrophoresis. To overcome the limits of detection, three amplification assays followed by hybridization of the products to specific viral probes were conducted. A variety of chimeric molecules were detected in low concentrations. Five species of chimeric molecules were characterized in blood, tumors, and feathers. Chimeric molecules were detected in 18 of 36 dually infected birds from the first trial and in 14 of 21 dually infected birds from the second trial. The findings show that, in four out of seven groups of the second trial, the chimeric molecule species persisted on passage. PMID- 12887190 TI - An enzyme-linked immunosorbent assay with the recombinant merozoite protein as antigen for detection of antibodies to Eimeria necatrix. AB - A cDNA library was constructed with Eimeria necatrix merozoite mRNA and immunologically screened by chicken sera against this parasite. One of the positive clones containing an insert of 879 nucleotides, pNP19, showed similarity to part of a published gene expressed in E. tenella merozoite by the homology search system. The inserted DNA was subcloned into baculovirus, and a 35-kD protein was expressed, purified, and used for the antigen in enzyme-linked immunosorbent assay (ELISA). Antibodies from the chickens vaccinated with the E. necatrix attenuated strain, Nn-P125, were detected from 14 days after vaccination by ELISA. The mean absorbance increased rapidly to a peak around 21 days after vaccination; thereafter, it began to decline. Even though some of the vaccinated chickens showed very low levels of antibody response to the recombinant protein 56 days after vaccination, they were protected against challenge with virulent strain of E. necatrix. The mean absorbances in sera from both vaccinated and nonvaccinated chickens highly increased 14 days after challenge. On the other hand, the antibody was not detected in ELISA when chickens were exposed to other Eimeria species such as E. tenella, E. acervulina, and E. maxima. These results demonstrate that this recombinant protein is suitable for detecting the specific antibody in chickens infected with both attenuated and virulent strains of E. necatrix. PMID- 12887191 TI - Pathogenesis of chicken-passaged Newcastle disease viruses isolated from chickens and wild and exotic birds. AB - The pathogenesis of six Newcastle disease virus (NDV) isolates recovered from chickens (Ckn-LBM and Ckn-Australia) and wild (Anhinga) and exotic (YN parrot, pheasant, and dove) birds was examined after the isolates had been passaged four times in domestic chickens. Groups of 10 4-wk-old specific-pathogen-free white leghorn chickens were inoculated intraconjunctivally with each one of the isolates. The infected birds were observed for clinical disease and were euthanatized and sampled at selected times from 12 hr to 14 days postinoculation or at death. Tissues were examined by histopathology, by immunohistochemistry (IHC) to detect viral nucleoprotein (IHC/NP), and by in situ hybridization to detect viral mRNA and were double labeled for apoptosis (terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling ([TUNEL] or IHC/caspase-3) and viral nucleoprorein (IHC/NP). Birds infected with the three low virulence viruses (Ckn-LBM, YN parrot, and Ckn-Australia) did not develop clinical disease. Microscopic lesions were observed only at the inoculation site and in organs of the respiratory system. The detection of viral nucleoprotein (N) was restricted to the inoculation site. The pheasant and dove isolates were highly virulent for chickens with marked tropism for lymphoid tissues, confirmed by the presence of large numbers of cells positive for viral N protein and viral mRNA. Viral N protein was detected early in the cytoplasm of cells in the center of the splenic ellipsoids. The apoptosis assays (TUNEL and IHC/caspase-3) showed increased apoptosis in the splenic ellipsoids as well. Apparently, apoptosis is an important mechanism in lymphoid depletion during NDV infection. PMID- 12887192 TI - Isolation and characterization of monoclonal antibodies against structural proteins of infectious laryngotracheitis virus. AB - Thirteen infectious laryngotracheitis virus (ILTV)-specific monoclonal antibodies (MAbs) were isolated after immunization of mice with purified infectious laryngotracheitis virions. On the basis of their reactions in western blot analyses of ILTV-infected cells, the MAbs were assigned to five different virus proteins or protein groups. Two of the viral target proteins could be identified after transient expression of cloned ILTV genes in eucaryotic cells. The MAbs of group II detected a 60-kD protein that was shown to be the ILTV homologue of herpes simplex virus type 1 (HSV-1) glycoprotein (g)C. The MAbs of group I reacted with the positional homologue of HSV-1 gJ, which is encoded by the open reading frame (ORF) 5 gene within the unique short genome region of ILTV. The ORF 5 gene product of ILTV was previously described as a 60-kD glycoprotein (gp60), whereas multiple protein bands with apparent molecular masses of 85, 115, 160, and 200 kD were identified in the present study. Immunoelectron microscopy revealed that both gC and gJ of ILTV are localized in the envelope of virus particles, whereas the 15-kD protein detected by the MAbs of group III presumably represents a tegument component. Immunofluorescence analyses of infected cells demonstrated that the epitopes of the gC- and gJ-specific MAbs are conserved in all tested ILTV isolates originating from different parts of the world and that these MAbs are also suitable for in situ antigen detection in tissues of ILTV infected chickens. The remaining ILTV-specific MAbs recognized viral proteins of 22 kD (group IV) and 38 kD (group V) that were not further characterized up to now. PMID- 12887193 TI - Molecular characterization of reticuloendotheliosis virus insertions in the genome of field and vaccine strains of fowl poxvirus. AB - Evidence of the widespread occurrence of reticuloendotheliosis virus (REV) sequence insertions in fowl poxvirus (FPV) genome of field isolates and vaccine strains has increased in recent years. However, only those strains carrying a near intact REV provirus are more likely to cause problems in the field. Detection of the intact provirus or REV protein expression from FPV stocks has proven to be technically difficult. The objective of the present study was to evaluate current and newly developed REV and FPV polymerase chain reaction (PCR) assays to detect the presence of REV provirus in FPV samples. The second objective was to characterize REV insertions among recent "variant" FPV field isolates and vaccine strains. With REV, FPV, and heterologous REV-FPV primers, five FPV field isolates and four commercial vaccines were analyzed by PCR and nucleotide sequence analysis. Intact and truncated REV 5' long terminal repeat (LTR) sequences were detected in all FPV field isolates and vaccine strains, indicating heterogeneous REV genome populations. However only truncated 3' LTR and envelope sequences were detected among field isolates and in one vaccine strain. Amplifications of the REV envelope and 3' LTR provided strong evidence to indicate that these isolates carry a near intact REV genome. Three of the four FPV vaccine strains analyzed carried a solo complete or truncated 5' LTR sequence, indicating that intact REV provirus was not present. Comparison of PCR assays indicated that assays amplifying REV envelope and REV 3' LTR sequences provided a more accurate assessment of REV provirus than PCR assays that amplify the REV 5' LTR region. Therefore, to differentiate FPV strains that carry intact REV provirus from those that carry solo 5' LTR sequences, positive PCR results with primers that amplify the 5' LTR should be confirmed with more specific PCR assays, such as the envelope, or the REV 3' LTR PCR. PMID- 12887194 TI - Evaluation of the non-temperature-sensitive field clonal isolates of the Mycoplasma synoviae vaccine strain MS-H. AB - The live attenuated temperature-sensitive (ts+) Mycoplasma synovia (MS) strain, MS-H, is used as a vaccine in a number of countries to control virulent MS infection in commercial chicken flocks. Nine out of 50 isolates made from flocks vaccinated with MS-H were found to have lost the ts+ phenotype of the original vaccine strain. In order to examine the influence of the ts- phenotype on virulence of the isolates, four of the ts- isolates, the MS-H vaccine, and the vaccine parent strain 86079/7NS were administered by aerosol in conjunction with infectious bronchitis virus to 3-wk-old specific-pathogen-free chickens. The four ts- clones induced only minimal air sac lesions that were not different in severity from those caused by MS-H vaccine; however, the vaccine parent strain 86079/7NS caused air sac lesions that were significantly greater than those of MS H and all ts- clones. The vaccine parent strain 86079/7NS and two of the ts- clones were recovered from the air sacs of the respectively infected chickens whereas the MS-H vaccine and two other ts- clones were not. Three of the ts- isolates caused increased tracheal mucosal thicknesses that were significantly greater than those from birds inoculated with MS-H, and one caused increased tracheal mucosal thicknesses that were significantly less than those from birds inoculated with 86079/7NS. In conclusion, unlike the MS-H vaccine, the MS-H ts- clones were associated with minor changes in tracheal mucosa; however, unlike the vaccine parent strain, they did not induce lesions in the air sacs. These results suggest that factors other than ts+ phenotype are involved in the attenuation of the MS-H vaccine. PMID- 12887195 TI - A longitudinal study of a novel dot-enzyme-linked immunosorbent assay for detection of avian influenza virus. AB - A monoclonal antibody (MAb)-based dot-enzyme-linked immunosorbent assay (ELISA) has been developed that detected the epitopes specifically associated with avian influenza virus (AIV). The dot-ELISA detected the antigens of AIV directly from clinical and field specimens. Data obtained from experimentally AIV-infected specific-pathogen-free chickens and also the 2001/02 AIV outbreak of serotype H7N2 positive flocks in Pennsylvania indicated that the mean sensitivity (Se) of the dot-ELISA ranged between 45% and 68% and the mean specificity (Sp), between 85% and 90%. The values were derived from various clinical and field specimens when compared with virus isolation with embryonating chicken eggs. On routine AIV surveillance samples, the dot-ELISA achieved a 92%-100% Sp on the basis of resting over 1500 AIV surveillance samples that were confirmed negative by virus isolation. The dot-ELISA detected AIV antigens with a 5-microl allantoic fluid sample that contained a concentration of 0.4 hemagglutinating units. Furthermore, the dot-ELISA retained its specificity for AIV because no cross-reactions were obtained with various other avian viruses. The findings in this study indicated that the dot-ELISA was highly sensitive and specific and comparable with the commercial Directigen test in the detection of AIV obtained from clinical and field specimens. PMID- 12887196 TI - Prediction of chicken embryo lethality with the avian Escherichia coli traits complement resistance, colicin V production, and presence of the increased serum survival gene cluster (iss). AB - Differentiating between virulent and avirulent avian Escherichia coli isolates continues to be a problem for poultry diagnostic laboratories and the study of colibacillosis in poultry. The ability of a laboratory to conduct one simple test that correlates with virulence would simplify studies in these areas; however, previous studies have not enabled researchers to establish such a test. In this study, the occurrence of certain phenotypic and genotypic traits purported to contribute to avian E. coli virulence in 20 avian E. coli isolates was correlated with the results of embryo challenge studies. This analysis was undertaken in an effort to determine which trait(s) best identified each avian E. coli isolate as virulent or avirulent. Traits selected were complement resistance, production of colicin V (ColV), motility, type F1 pili expression, presence of the temperature sensitive hemagglutinin gene (tsh), and presence of the increased serum survival genetic locus (iss). ColV production, complement resistance, and presence of the iss genetic element were the three traits most highly correlated with high embryo lethality. A logistic regression model was used to predict the embryo lethality results on the basis of the most frequent isolate characteristics. Results indicate that ColV, complement resistance, and if are significant predictor variables for the percentage of embryo lethality resulting from challenge with a specific avian E. coli isolate. However, no single trait has the ability to predict virulent isolates 100% of the time. Such results suggest the possibility that the embryo lethality assay may prove to be the one test needed to determine if an avian E. coli isolate is virulent. PMID- 12887197 TI - Rapid detection of Salmonella from poultry by real-time polymerase chain reaction with fluorescent hybridization probes. AB - Detection of Salmonella by bacteriologic methods is known to be time consuming. Therefore, we have developed a real-time probe-specific polymerase chain reaction (PCR) to rapidly detect Salmonella invA gene-based PCR products from chicken feces and carcasses by a fluorescence resonance energy transfer assay. The sensitivity and the specificity of this system were determined as 3 colony forming units ml(-1) and 100%, respectively. Overnight tetrathionate broth enrichment cultures of chicken feces and carcass samples were used in template preparation for PCR. Also, a standard bacteriology was performed (National Poultry Improvement Plan-U.S. Department of Agriculture, Bacteriological Analytical Manual-Food and Drug Administration Center for Food Safety and Applied Nutrition) for confirmation. Seventy-two cloacal swab, 147 intestine, and 50 carcass (neck) samples were examined. Thirteen (8.8%) and 25 (17%) of the intestinal samples were found to harbor Salmonella by bacteriology and PCR, respectively. Forty-five of 50 (90%) carcass samples were Salmonella positive by both methods. Salmonella was not detected from cloacal swab samples. Results indicate that this assay has the potential for use in routine monitoring and detection of Salmonella in infected flocks and carcasses. PMID- 12887198 TI - A restriction fragment length polymorphism-based polymerase chain reaction as an alternative to serotyping for identifying Salmonella serotypes. AB - The phase 1 (fliC) and phase 2 (fljB) Salmonella flagella genes were analyzed by restriction fragment length polymorphism (RFLP)-polymerase chain reaction (PCR) to aid in the identification of different Salmonella serotypes. Twenty-four phase 1 flagellin and eight phase 2 flagellin genes could be differentiated among each other with restriction endonucleases Sau3A and HhaI in RFLP-PCR analysis. These flagellin genes comprise the major antigenic formulas for 52 serotypes of Salmonella sp., which include the common serotypes found in poultry and other important food animal species. With the knowledge of the O antigen composition determined from conventional O serotyping, 90% of the Salmonella serotypes could be identified by this double restriction enzyme RFLP analysis of fliC and fljB genes. This RFLP-PCR flagellar typing scheme was successfully applied to the identification of serotype for 112 Salmonella isolates obtained from poultry environment. There was a significant correlation between RFLP-PCR and conventional serotyping (chi-square, P < 0.001). Overall, PCR-RFLP proved to be a fast, accurate, and economical alternative approach to serotyping Salmonella sp. PMID- 12887199 TI - Enhancement of enteropathogenic Escherichia coli pathogenicity in young turkeys by concurrent turkey coronavirus infection. AB - In a previous study, turkey coronavirus (TCV) and enteropathogenic Escherichia coli (EPEC) were shown to synergistically interact in young turkeys coinfected with these agents. In that study, inapparent or mild disease was observed in turkeys inoculated with only TCV or EPEC, whereas severe growth depression and high mortality were observed in dually inoculated turkeys. The purpose of the present study was to further evaluate the pathogenesis of combined TCV/EPEC infection in young turkeys and determine the role of these agents in the observed synergistic interaction. Experiments were conducted to determine 1) effect of EPEC dose, with and without concurrent TCV infection, and 2) effect of TCV exposure, before and after EPEC exposure, on development of clinical disease. Additionally, the effect of combined infection on TCV and EPEC shedding was determined. No clinical sign of disease and no attaching and effacing (AE) lesions characteristic of EPEC were observed in turkeys inoculated with only EPEC isolate R98/5, even when turkeys were inoculated with 10(10) colony forming units (CFU) EPEC (high dose exposure). Only mild growth depression was observed in turkeys inoculated with only TCV; however, turkeys inoculated with both TCV and 10(4) CFU EPEC (low dose exposure) developed severe disease characterized by high mortality, marked growth depression, and AE lesions. Inoculation of turkeys with TCV 7 days prior to EPEC inoculation produced more severe disease (numerically greater mortality, significantly lower survival probability [P < 0.05], increased frequency of AE lesions) than that observed in turkeys inoculated with EPEC prior to TCV or simultaneously inoculated with these agents. Coinfection of turkeys with TCV and EPEC resulted in significantly increased (P < 0.05) shedding of EPEC, but not TCV, in intestinal contents of turkeys. These findings indicate that TCV infection predisposes young turkeys to secondary EPEC infection and potentiates the expression of EPEC pathogenicity in young turkeys. PMID- 12887200 TI - Genotype analyses of Campylobacter isolated from the gastrointestinal tracts and the reproductive tracts of broiler breeder roosters. AB - Campylobacter is considered to be the leading bacterial etiologic agent of acute gastroenteritis in humans. Evidence implicates poultry as a major source of the organism for human illness; however, the pathways involved in Campylobacter contamination of poultry flocks, horizontal transmission and/or vertical transmission, remain unclear. Recent evidence implicates breeders as a potential source for Campylobacter contamination of the subsequent broiler offspring. In this investigation, Campylobacter isolated from feces, cloacal swabs, ceca, semen, and vas deferens of 12 breeder broiler roosters were genotyped by both flagellin A short variable region (flaA SVR) DNA sequence analysis and repetitive element (rep)-polymerase chain reaction (PCR). In 9 of 12 roosters, Campylobacter was isolated from multiple sites sampled. Comparison of multiple isolates obtained from individual roosters revealed variable results. In five of the nine roosters, all Campylobacter isolated demonstrated closely related flaA SVR DNA sequences as well as rep-PCR patterns; isolates from these roosters were collected from both the gastrointestinal and the reproductive tracts or from the gastrointestinal tract alone. The remaining four roosters had Campylobacter that were distinct by both typing methods. Isolates from two of these four roosters originated from both the gastrointestinal and the reproductive tracts. Isolates from the remaining two roosters originated from only the reproductive tract. Comparisons of all Campylobacter isolates recovered from a distinct sample type within either the reproductive tract or the gastrointestinal tract (feces, semen, cloaca, vas deferens, or ceca) were quite diverse. No relationship between the genotypes and the sample type could be ascertained. Further investigation is needed to determine the route of contamination and if the presence of Campylobacter within the rooster leads to contamination of the broiler offspring via the fertilized egg. PMID- 12887201 TI - Effect of ochratoxin A on Escherichia coli-challenged broiler chicks. AB - A study was conducted to evaluate the effects of ochratoxin A (OA) on Escherichia coli-challenged broiler chickens. Day-old broiler chicks were separated into two groups of 92 chicks each, with one group fed a control mash diet, and the other fed a mash diet containing 2 ppm OA. On day 14, each group was further separated into two groups, with one group inoculated with E. coli O78 (1 x 10(7) colony forming units/0.5 ml), whereas the other group was not inoculated with E. coli. After E. coli inoculation on day 14, four birds from each group were euthanatized at 1, 2, 3, 5, 7, 10, 14, and 21 days postinoculation. Escherichia coli infection caused dullness, depression, huddling, and diarrhea. Mortality was 14.3% in chicks infected with E. coli but fed no OA. Mortality increased to 35.7% in chicks fed OA and infected with E. coli. Decreased body weight and reduced feed intake were observed in chicks fed OA, and the effects were more pronounced in chicks fed OA and infected with E. coli. Increased serum levels of aspartate aminotransferase, alanine aminotransferase, uric acid, and creatinine and decreased levels of total proteins, albumin, globulins, calcium, and phosphorus were observed in OA-fed birds. Escherichia coli infection did not cause significant alteration in any of the serum biochemical parameters. The presence of OA in poultry rations increased mortality and the severity of an E. coli infection. PMID- 12887202 TI - Pathogenicity of two recombinant avian leukosis viruses. AB - We have recently described the isolation and molecular characteristics of two recombinant avian leukosis subgroup J viruses (ALV J) with an avian leukosis virus subgroup A envelope (r5701A and r6803A). In the present study, we examined the role of the subgroup A envelope in the pathogenesis of these recombinant viruses. Chickens of line 151(5) x 7(1) were inoculated at 1 day of age with r5701A, r6803A, Rous-associated virus type 1 (RAV-1), or strain ADOL-Hcl of ALV J. At 2, 4, 10, 18, and 32 wk postinoculation (PI), chickens were tested for avian leukosis virus (ALV)-induced viremia, shedding, and neutralizing antibodies. All except one chicken inoculated with the recombinant viruses (98%) developed neutralizing antibodies by 10 wk PI compared with only 16% and 46% of the ADOL-Hcl and RAV-1-inoculated birds, respectively. ALV-induced tumors and mortality in the two groups inoculated with recombinant viruses were different. The incidence of tumors in groups inoculated with r5701A or RAV-1 was 100% compared with only 9% in the groups inoculated with r6803A or ADOL-Hcl. The data suggest that differences in pathogenicity between the two recombinant viruses might be due to differences in the sequence of the 3' untranslated region (presence or absence of the E element), and, therefore, not only the envelope but also other elements of the viral genome play an important role in the pathogenesis of ALV. PMID- 12887203 TI - A model of avian mycobacteriosis: clinical and histopathologic findings in Japanese quail (Coturnix coturnix japonica) intravenously inoculated with Mycobacterium avium. AB - Mycobacterial infections are an important cause of morbidity and mortality in birds and a considerable diagnostic challenge until the disease is advanced. In order to develop more clinically useful antemortem tests, a biological model was created that replicated naturally occurring disease. Japanese quail (Coturnix coturnix japonica; n = 8) were inoculated intravenously with Mycobacterium avium. Two additional birds served as uninoculated controls. Mean survival time of the inoculated birds was 68 +/- 13 days postinoculation (PI). Seven of the eight inoculated birds died naturally. Clinical and postmortem abnormalities in inoculated birds were characteristic of naturally occurring mycobacteriosis. Abnormal clinical findings included decreased activity, feather erection, and sudden death. Mean body weight and packed cell volume declined and mean total white blood cells (primarily heterophils, bands, and monocytes) increased from 28 days PI onward. Similar to birds that are naturally infected with mycobacteriosis, the inoculated birds were thin and had severe hepatosplenomegaly on postmortem examination. All eight birds had lesions in the liver, spleen, intestine, lung, gonads, and serosa. Less commonly affected tissues included bone marrow, thymus, gizzard, heart, pancreas, and brain. Lesions were invariably severe in the liver and spleen. These gross postmortem findings were consistent with natural infections of avian mycobacteriosis. Mycobacterium avium was isolated from the liver, spleen, and intestine of all inoculated birds. Both control birds remained disease free and culture negative. This inoculation protocol is a reliable and practical means of inducing avian mycobacteriosis for further study. PMID- 12887204 TI - Diagnosis of avian mycobacteriosis: comparison of culture, acid-fast stains, and polymerase chain reaction for the identification of Mycobacterium avium in experimentally inoculated Japanese quail (Coturnix coturnix japonica). AB - In this study we compared culture, acid-fast stains, and polymerase chain reaction (PCR) for the detection of acid-fast organisms in fecal and tissue samples from Japanese quail (Coturnix coturnix japonica) that were experimentally inoculated intravenously with Mycobacterium avium. For culture, three different culture media (modified Herrold egg yolk with mycobactin; Lowenstein-Jensen [L J]; and L-J with cyclohexamide, naladixic acid, and lincomycin) were tested to determine which medium had the greatest success in isolating mycobacteria. Acid fast staining methods included Zichl-Neelsen (Z-N) and Truant. The PCR assay detected mycobacterial DNA with primers specific for the 65-kD heat shock protein gene. Culture was considered the "gold standard." Compared with other culture media, L-J yielded more positive cultures and greater numbers of colonies on positive tubes, and incubation times were shorter. Mycobacterium avium was isolated from all of the harvested tissue samples (liver, spleen, and intestine) of inoculated birds. Mycobacteria were isolated from 53% (69/130) of fecal samples from inoculated birds. As the disease advanced, fecal culture was positive on more culture days, indicating that the culture-positive rate was higher later in the course of the disease. Compared with culture, all of the laboratory methods had 100% specificity for the tissue samples. Sensitivities for the tissue samples were 82.6% (Z-N), 95.7% (Truant), and 100% (PCR). For the fecal samples, the specificity was >95% for all methods. Sensitivities compared with fecal culture were 7.2% (Z-N), 30.4% (Truant), and 20.3% (PCR). Tissue and fecal samples from the two control birds were negative for acid-fast organisms by any method. These results were comparable with clinical cases of avian mycobacteriosis where culture and PCR of tissue samples seem to be the most sensitive and specific laboratory tests and evaluation of fecal samples still remains challenging. On the basis of the results of this study, identification of mycobacteria in fecal samples from Japanese quail can be optimized by repeated cultures and Truant acid-fast staining of fecal smears. PMID- 12887205 TI - Detection of antibody-forming cells directed against Newcastle disease virus and their immunoglobulin class by double immunoenzyme histochemistry. AB - Antibody-forming cells (AFCs) against Newcastle disease virus (NDV) and their immunoglobulin (Ig) class were demonstrated by a double immuno-enzyme histochemical technique. The AFCs were stained and quantified in spleen sections of chickens euthanatized at day 7 postexposure to the Roakin strain of NDV. The sections were incubated with NDV to determine the specificity of the AFCs. Bound virus was subsequently visualized with a primary monoclonal antibody (MAb), a secondary horseradish peroxidase-conjugated MAb, and 3-amino-9-ethylcarbazole as substrate. IgM and IgA were stained with MAbs and an alkaline phosphatase (AP) conjugated secondary antibody. IgG class antibodies were demonstrated with an AP conjugated rabbit serum. The final substrate for the three Igs was naphthol AS-MX phosphate and fast blue BB. About 64-159/mm2 AFCs against NDV were detected. Of these virus-binding cells, about 55% produced IgM, 37% produced IgG, and the remaining 8% produced IgA. PMID- 12887206 TI - Mycoplasma gallinarum infection in commercial layers and onset of fatty liver hemorrhagic syndrome. AB - Fatty liver hemorrhagic syndrome (FLHS) was observed in each of three trials in which commercial layers were utilized to determine the effect of Mycoplasma gallinarum (MGn) on egg and eggshell quality parameters and egg production. In each of three trials, FLHS occurred 31-54 days later in MGn-inoculated hens as compared with the Mycoplasma-clean (control) hens. In trials 1 and 2, no therapeutic intervention was initiated to ameliorate FLHS. In trial 3, therapeutic intervention was instituted and consisted of the addition of 1 pound of choline chloride/ton of feed. Total mortality recorded throughout the duration of each trial and attributable to FLHS was not significantly different between the control and the MGn-inoculated treatment. However, FLHS-associated mortality in each of the three trials was numerically greater for the control treatment. PMID- 12887207 TI - Coccidiosis control with toltrazuril in conjunction with anticoccidial medicated or nonmedicated feed. AB - A 42-day broiler floor pen study was conducted comparing the anticoccidial efficacy of toltrazuril (Baycox) as a stand alone treatment and as an additional treatment to in-feed anticoccidial programs. Toltrazuril was administered on days 18 and 19 in the drinking water at 7 mg/kg of body weight. The treatments were 125 ppm nicarbazin (days 0-14) to 66 ppm salinomycin (SAL) (days 15-35) with and without toltrazuril, SAL (days 0-35) with and without toltrazuril, nonmedicated (NM) to SAL with toltrazuril, and NM with and without toltrazuril. The controls were NM noninfected and infected. The treatments were replicated in five blocks of eight pens each in a randomized complete block design. All withdrawal feed was nonmedicated. On day 14, birds, except noninfected, were exposed to coccidial oocysts (Eimeria acervulina, Eimeria maxima, and Eimeria tenella) seeded litter. On days 21, 28, 35, and 42, birds and feed were weighed, four birds per pen were coccidial lesion scored, and litter oocyst counts were performed. The coccidial infection in the NM infected treatment caused a significant (P < 0.05) coccidiosis infection. Coccidiosis was moderately controlled in the anticoccidial treatment birds without toltrazuril. Performance in the NM with toltrazuril was equal to or better (P < 0.05) than the anticoccidial programs without toltrazuril. Toltrazuril was equal to the noninfected birds in performance. Toltrazuril most completely eliminated all coccidial lesions and dramatically reduced oocyst shedding. The performance data, lesion scores, and oocyst counts showed that a 2-day treatment with toltrazuril successfully controlled the coccidiosis with no relapse of infection. Toltrazuril can thus be used for supplemental control with in-feed anticoccidials or as a primary anticoccidial with nonmedicated feed. PMID- 12887208 TI - Experimental inoculation of pigeons (Columba livia) with Mycobacterium bovis. AB - The purpose of this pilot study was to determine if pigeons (Columba livia) are susceptible to infection with Mycobacterium bovis by either oral or intratracheal inoculation and to assess their possible role in the lateral transmission of bovine tuberculosis. Six pigeons were orally inoculated with 1.3 x 10(5) colony forming units of M. bovis, six pigeons were intratracheally inoculated with the same dose, and six pigeons served as noninoculated controls. The study continued for 90 days postinoculation (PI), with groups of birds necropsied at 30-day intervals, and fecal samples and tissues were collected for mycobacterial culture. Two pigeons, one intratracheally inoculated and one orally inoculated, shed M. bovis in their feces at 1 day PI, and one intratracheally inoculated bird shed M. bovis in its feces 60 days PI. Whereas no illness or weight loss was present during the course of the study, 2 of 12 inoculated birds exhibited microscopic lesions of mycobacteriosis, and the organism was isolated from tissues of three inoculated birds. Pigeons are susceptible to infection with M. bovis after high dose inoculation and can shed the organism in their feces for up to 60 days PI; intratracheally inoculated birds appear more likely to become active fecal shedders of M. bovis. Although these were high dose inoculations under experimental conditions, pigeons may potentially play a role in the lateral transmission of bovine tuberculosis between infected and uninfected mammalian hosts. PMID- 12887209 TI - In vitro susceptibility of Ornithobacterium rhinotracheale to several antimicrobial drugs. AB - As part of the basic characterization of Ornithobacterium rhinotracheale, the minimal inhibitory concentrations of 10 antimicrobial drugs were determined for reference strains and Mexican isolates by a broth microdilution method. For optimal growth of the organisms, a supplemented brain-heart infusion broth was used. The susceptibility of O. rhinotracheale to amoxicillin, enrofloxacin, and oxytetracycline was variable. However, consistent higher minimal inhibitory concentrations values were obtained for gentamicin, fosfomycin, trimethoprim, sulfamethazine, sulfamerazine, sulfaquinoxaline, and sulfachloropyridazine. Obtained results among Mexican isolates indicate a marked antimicrobial drug resistance trend. PMID- 12887210 TI - Presence of avian pneumovirus subtypes A and B in Japan. AB - Four avian pneumovirus (APV) isolates from chickens clinically diagnosed with swollen head syndrome were genetically characterized as to the subtypes of the virus in Japan. The results of reverse transcriptase-polymerase chain reactions based on subtype-specific primers and direct sequence analysis of G genes indicated subtypes A and B but not C or D of APV were present in Japan. Several routes or sources are conceivable for APV to invade into Japan. PMID- 12887211 TI - Mycoplasma gallisepticum-induced alterations in chicken red blood cells. AB - Incubation of Mycoplasma gallisepticum with washed chicken red blood cells for 1 hr or 5 hr resulted in altered red blood cell surface morphology and perforations of the cells. PMID- 12887212 TI - Direct lateral transmission of Histomonas meleagridis in turkeys. AB - The lateral transmission of Histomonas meleagridis in turkeys was studied in floor pens without the presence of Heterakis gallinarum. Battery-reared poults (120) were transferred at 2 wk of age to concrete-floored floor pens with fresh pine shavings litter (40/group). One group received no exposure. In other groups, either 10% or 25% of the birds were inoculated per cloaca with cultured H. meleagridis (200,000/bird) and placed in the pens as seeder birds. Inoculated birds died at 10-18 days postinfection (PI) showing typical liver and cecal lesions of histomoniasis. Birds in the high-exposure group died of histomoniasis beginning 16 days PI and continuing to 100% mortality by day 23 PI. Birds in the low-exposure (LE) group died beginning on day 19 PI and continuing through day 31 PI. All but one LE bird alive on day 31 PI had severe liver and cecal lesions of histomoniasis at necropsy. There was no evidence of histomoniasis in unexposed birds. No cecal worms (H. gallinarum) were found at necropsy of dead birds or in unexposed birds at the end of the experiment. Even though H. gallinarum is the only known reservoir for H. meleagridis, these results suggest that lateral transmission of histomoniasis through a flock can occur readily through normal contact between uninfected birds and infected birds and their droppings in the total absence of cecal worms. PMID- 12887213 TI - Outbreak of salmonellosis in a zoologic collection of lorikeets and lories (Trichoglossus, Lorius, and Eos spp.). AB - During August 2001, a syndrome characterized by acute lethargy and dyspnea was observed in a population of 45 lorikeets and lories in an open-air zoologic exhibit. The first death occurred on August 10, and within the next 12 days, nine more birds died (22% mortality rate). Hepatomegaly, reddening and congestion of the lungs, and injection of the serosal surface of the intestines were the common gross findings. Histologic changes, including fibrinonecrotic hepatitis and splenitis, bacterial emboli (liver, spleen, lung, kidney, proventriculus), pulmonary congestion and hemorrhage, and enteritis, were indicative of an acute, overwhelming bacterial septicemia. Salmonella typhimurium, with the same antibiogram, was isolated from four birds. Several birds had attacked and killed a snake on July 24, and Salmonella serogroup B (untypeable) was isolated from intestine and kidney samples of a garter snake caught in the open-air exhibit on August 28. Salmonella was also isolated from environmental samples of the exhibit but not from food preparation areas. After antimicrobial therapy, Salmonella spp. was not isolated from the surviving birds. The source of Salmonella in this outbreak remains unknown, but infection either directly or indirectly from snakes in the exhibit is possible. Contact between captive psittacine populations and reptiles should be avoided to prevent the risk of salmonellosis. PMID- 12887214 TI - Isolation and characterization of a 6/85-like Mycoplasma gallisepticum from commercial laying hens. AB - Eighty-three-week-old table egg layers with swollen sinuses were presented with a history of increased mortality. Serology revealed positive titers to Mycoplasma gallisepticum (MG). The birds were part of a flock in which some birds had been vaccinated with 6/85 live MG vaccine at 18 wk of age. Tracheal cultures were obtained from both vaccinated and unvaccinated birds within the flock. The cultures were indistinguishable from 6/85 vaccine by both random amplified polymorphic DNA analysis and DNA sequence analysis. Challenge studies were performed to compare the field isolates with 6/85 vaccine and the R strain of MG. The field isolates produced a greater antibody response by serum plate agglutination than did the 6/85 vaccine. The isolates effectively colonized the trachea without increasing the tracheal mucosal thickness; however, they did not extensively colonize the air sacs or cause airsacculitis in the experimental birds. PMID- 12887215 TI - Chaunocephalus ferox in free-living white storks in central Spain. AB - Ten out of 42 (23.8%) white storks (Ciconia ciconia) admitted to two rehabilitation centers in central Spain had lesions caused by the trematode Chaunocephalus ferox in the small intestinal wall. Fourteen of the examined birds were adults, five were subadults, and 23 were chicks of various ages. Parasitation was 32% (n = 8) in chicks and 13% (n = 2) in adult birds, whereas no juvenile bird was affected. Among dead birds, stork chicks affected by C. ferox lesions had a lower body weight (2196.1 g, SD = 814.2) than storks without lesions (2965.8 g, SD = 742.9, P < 0.05). Two chicks were additionally infected with Salmonella subspecies I serotype enteritidis 1,9,12: g, m:1, 7. Prevalence of the parasite in the examined birds was lower than in a population of Asian open-billed storks (Anastomus oscitans), in which it was pathogenic due to the destruction of the tunica muscularis and formation of large granulomatous lesions in the wall of the postduodenal portion of the small intestine. Pathogenic alterations caused by C. ferox are presumed to be related to numbers of adults present. Because storks admitted to rehabilitation centers suffer stress due to various reasons that may lower their immune response and exacerbate existing infections, the analysis of fecal sediments of white storks admitted for rehabilitation is recommended. PMID- 12887216 TI - Natality and the changing pattern of seasonality of births in the province of Teramo (Abruzzo, Italy: 1500-1871). AB - Reconstruction of human ecosystems and their stability over time provides knowledge of the processes of adaptability developed by isolated communities. Seasonality of vital events is a good indicator of the effects of different 'traditional' lifestyles, which in turn depend on the ecological context in which a population developed specific subsistence models. Seasonality of births reflects the cultural attitude towards the best time to conceive, in relation to work activities and loads; the latter may also affect physiological functions related to fertility. The present research concerns gross birth rates and seasonality of births and conceptions during four centuries in south-central Italy. Birth rates were between 33.0 and 36.5 per 1000. The pattern of seasonality of births and, by extension, of conceptions defines a southern-type agricultural area for the earlier periods. However, it also shows a progressive shift towards an increasing concentration of conceptions in spring-summer--namely from April to August--with a large increase in summer in the 19th century with respect to the previous periods. The new 19th century pattern is reported by Crisafulli, Dalla Zuanna & Solero (2000) as being representative of the central Adriatic region, a geographical classification to which Abruzzo can also be attributed. PMID- 12887217 TI - An analysis of childhood malnutrition in Kerala and Goa. AB - Improved child health and survival are considered universal humanitarian goals. In this respect, understanding the nutritional status of children has far reaching implications for the better development of future generations. The present study assessed, first, the nutritional status of children below 5 years using the three anthropometric measures weight-for-age, height-for-age and weight for-height in two states of India, Kerala and Goa. Secondly, it examined the confounding factors that influence the nutritional status of children in these states. The NFHS-I data for Kerala and Goa were used. The results showed that the relative prevalence of underweight and wasting was high in Kerala, but the prevalence of stunting was medium. In Goa, on the other hand, the relative prevalence of wasting and underweight was very high, and that of stunting was high. Both socioeconomic and family planning variables were significantly associated with malnutrition in these states, but at varied levels. The study recommends more area-specific policies for the development of nutritional intervention programmes. PMID- 12887218 TI - Analysing cross-sectional data with time-dependent covariates: the case of age at first birth in South Africa. AB - Analysing time-dependent independent variables requires the use of process oriented statistical models. Yet social scientists--especially those in poor countries--have often had to use data collected at a single point in time, making their task difficult. Making several assumptions about the covariates, the present study uses survival analysis and other statistical techniques to analyse the 1996 South African population census data and examine the effects of selected independent variables on the timing of parenthood in the country. It was found that the onset of parenthood occurs late in South Africa compared with the pattern in most other African societies. While education plays a role in the postponement of parenthood within racial groups, it fails to explain the differences between African and Coloured women on the one hand, and White and Asian women on the other hand, a finding that suggests the existence of two regimes of family formation in South African society. PMID- 12887219 TI - Patrilineal family values, family planning and variation in stature among Taiwanese six-year-olds. AB - It has been argued that patrilineal joint family systems tend to bias family planning decisions in favour of sons. A simple model suggests that in such societies, any given son will be more highly valued by his parents (1) the fewer his brothers and (2) the earlier his birth is in the brother series. A daughter's value will be greater (1) the fewer brothers she has and (2) the earlier her birth is relative to other sisters. This study first addresses the extent of son preference as inferred from family composition data for 772 Taiwanese first graders born in the mid-1970s in two socioeconomically distinct communities in Taipei, Taiwan. It then uses linear regression to consider whether the model criteria help account for statural variation among children in each study area when controlling for differences in measurement age, parental education and housing. With respect to family composition and gender preference, available evidence was consistent with previous surveys. While better-educated parents in the more affluent study area had significantly fewer children (p < 0.0005) and were more willing to stop without a son, girls there, as in the less affluent area, were still significantly more likely than boys to belong to large sibships (p < or = 0.005). Evidence from mean height of males and females partially accords with hypothetical predictions. In the less affluent area, the interaction effect of male birth order and the presence of younger siblings was significantly associated with mean stature (p = 0.002). Males without brothers were 2.0 cm taller than males with either an older or a younger brother (116.3 +/- 0.5 cm vs 114.3 +/- 0.4 cm). Males who had both younger and older brothers, but often no sisters, were about as tall, however, as those without brothers. A similar, but less pronounced, pattern was found among males in the more affluent area, but only among those who had sisters. These boys were also consistently shorter than boys without sisters (115.6 +/- 0.6 cm vs 117.7 +/- 0.6 cm; p = 0.001). Patterns of mean female stature did not clearly support the hypothesis. Girls in the more affluent area were relatively tall and did not show significant variation. Results among less affluent girls showed significant contrasts, but not necessarily in the predicted direction. PMID- 12887220 TI - Perception of risk of HIV/AIDS and sexual behaviour in Kenya. AB - The association between perception of risk of HIV infection and sexual behaviour remains poorly understood, although perception of risk is considered to be the first stage towards behavioural change from risk-taking to safer behaviour. Using data from the 1998 Kenya Demographic and Health Survey, logistic regression models were fitted to examine the direction and the strength of the association between perceived risk of HIV/AIDS and risky sexual behaviour in the last 12 months before the survey. The findings indicate a strong positive association between perceived risk of HIV/AIDS and risky sexual behaviour for both women and men. Controlling for sociodemographic, sexual exposure and knowledge factors such as age, marital status, education, work status, residence, ethnicity, source of AIDS information, specific knowledge of AIDS, and condom use to avoid AIDS did not change the direction of the association, but altered its strength slightly. Young and unmarried women and men were more likely than older and married ones to report risky sexual behaviour. Ethnicity was significantly associated with risky sexual behaviour, suggesting a need to identify the contextual and social factors that influence behaviour among Kenyan people. PMID- 12887221 TI - A re-examination of levels and differential in fertility in South Africa from recent evidence. AB - The final estimate of South Africa's population as of October 1996 from the first post-apartheid census by Statistics South Africa was lower (40.6 million) than expected (42 million). The expectation of a total population of 42 million was largely based on results of apartheid projections of South Africa's population. The results of the last apartheid census in South Africa in 1991 had been adjusted such that it was consistent with results modelling the population size of South Africa. The discrepancy between the final estimate of the 1996 census and that expected from the modelling described above, and the departure by Statistics South Africa from previous practice of adjusting the census results to be consistent with demographic models, has generated controversies regarding the accuracy of the final results from the 1996 census. This study re-examines levels and differential in fertility in South Africa from recent evidence in order to assess whether or not the fertility inputs in projections of South Africa's population during the apartheid era overestimated fertility. PMID- 12887222 TI - Social linkages to biological markers of health among the elderly. AB - The social environment and exposure to life challenge affect a person's physical and emotional well-being. The present research uses a population-based study of the elderly in Taiwan to elaborate the cumulative physiological costs--as reflected in biological markers of risk factors known to have adverse consequences for health--of challenge and unfavourable position in social hierarchies and networks. Overall, biological markers of risk among the elderly are similar in Taiwan and the United States. However, male and female Taiwanese elderly are at lower risk for illness associated with indicators of DHEA-S, while women are at higher risk for illness associated with elevated blood pressure, and men at lower risk for illness associated with total/HDL cholesterol, and glycosylated haemoglobin. There are strong and statistically significant effects of position in social hierarchy (education) and challenge (recent widowhood and a perception of high demands) on an index of cumulative risk (allostatic load). Membership in social networks and participation in social activities have expected, but not statistically discernible, effects. PMID- 12887223 TI - The influence of birth order on birth weight: does the sex of preceding siblings matter? AB - The purpose of this study was to investigate whether the well-established relationship between parity and birth weight is affected by the sex composition of siblings, especially for male newborns. Subjects were 856 male and 862 female newborns who weighed at least 2500 g at birth, who were born after 37 completed weeks of gestation, who obtained an Apgar score of 7 or higher, who had the same biological parents as all other children in the sibship, and who lived in the same household. Information on birth weight was collected from hospital records. Results showed that male newborns with older brothers weighed less than male newborns with older sisters. In contrast, the weight of female newborns with older brothers did not differ from the weight of female newborns with older sisters. One explanation of these results is that maternal immunoreactivity to some male-specific feature of the fetus affects prenatal development and consequently reduces birth weight in males. The relation between older brothers and birth weight may have theoretical significance for behavioural variables. PMID- 12887224 TI - Religiosity and its relation to blood pressure among selected Kuwaitis. AB - This study examines the relationship between blood pressure and the religious practices of Kuwaitis as members of a Muslim society. Religious variables were measured via a sociocultural questionnaire. Blood pressure measurements were taken with a sphygmomanometer. Non-opportunistic samples were taken from 223 Kuwaitis. The difference in religious commitment between Muslim Sunnis and Muslim Shiites was examined using a t-test. Matrix correlation was used to examine the relationship between religious commitment and some other variables. Multiple regression was conducted to determine the effect of religiosity on blood pressure, as well as statistically controlling for other variables such as body mass index, socioeconomic status, smoking, gender and age. It was found that both systolic and diastolic blood pressure were affected by religious commitment and religious activities PMID- 12887225 TI - Physical assessment of the bariatric person. PMID- 12887226 TI - Evidence-based nursing practice: to infinity and beyond. AB - BACKGROUND: Medical errors are in the news almost daily, frequently after patients have experienced ineffective and unsafe protocols. As a result, patients and family members lose trust in the healthcare system. Nursing practice is changing almost daily with advances in nursing research and improvements in technology and practice. Reading, understanding, and managing these large amounts of information have become difficult for most nurses and organizations. METHODS: This article provides an historical background for evidence-based practice and methods for assimilating research into practice. RESULTS: Systematic reviews and other decision-making models are discussed using specific questions for establishing policy guidelines. CONCLUSION: Evidence-based practice encompasses implementing the best-known practices into the clinical setting using a scientific approach. As a result, safe, high-quality, and cost-effective care will be more likely to occur consistently. PMID- 12887227 TI - Meeting the needs of the health system: a refresher course for registered nurses. AB - To address the shortage of nurses experienced by a healthcare system, faculty from a baccalaureate nursing program collaborated with the affiliated healthcare system members to offer a refresher course to facilitate the return of inactive nurses to the workforce. The program included didactic content, simulated laboratory experiences, and precepted clinical experiences on the employing units. This article describes the scope of the program and focuses on recommendations for future course implementation, including implementing a systematic evaluation of each participant's knowledge base at the beginning of the course and an extended course time frame to allow for greater integration of content. PMID- 12887228 TI - Developing and evaluating patient education materials. AB - Nurses should be involved in all aspects of patient education, including the development of print and web resources, but most nurses have not been educated in how to develop these resources. This article discusses the rationale for nurse involvement, describes guidelines for evaluating existing material for credibility and readability, and provides recommendations for rewriting material at an easier-to-read level. PMID- 12887229 TI - Developing a postbasic gerontology program for international learners: considerations for the process. AB - BACKGROUND: A private university in Chile was seeking opportunities to acquire expertise in developing distance education programs using new media technology, especially in health-related fields. An initiative was undertaken to develop a distance postbasic gerontology program for technical nurses in Chile by partnering with a Canadian school of nursing. METHOD: The instructional design process was used as a systematic approach throughout the development of the program. Attention to cultural considerations was emphasized throughout each phase of the process. RESULTS: The developed program consists of 13 self-paced print modules with web-based components and is primarily asynchronous with scheduled milestones. A 6-week preceptored clinical experience follows completion of the theory portion to support learner mastery of program outcomes. CONCLUSION: Developing international nursing education programs is an enriching experience that presents unique challenges and issues. These challenges can be overcome by using a creative, systematic approach to program development that reflects cultural considerations. Developing flexible working relationships that respect the cultural differences of international team members facilitates international initiatives. PMID- 12887230 TI - Comparison of oncology nurse and physician use of the Internet for continuing education. AB - BACKGROUND: In the past 10 years, there has been a dramatic increase in published literature on the topic of Internet use by healthcare professionals. Learners and organizations providing grants to the continuing education (CE) division of a medical education and publishing company have expressed increasing interest in the area of Internet CE. To quantify and analyze this trend, the company began using surveys to evaluate this growing area of interest. METHOD: Since 1998, annual surveys at the annual meetings of the Oncology Nursing Society and the American Society of Clinical Oncology were used to determine trends in educational use of the Internet for CE. FINDINGS: Internet use and frequency of use, including use of the Internet for CE among both nurses and physicians, have been increasing. When asked what would make these professionals more likely to use the Internet for CE, both indicated making the activities low cost or free and easier access. CONCLUSIONS: Nurses and physicians are looking to the Internet for quick access to reliable information, thus challenging CE educators to develop ways to implement Internet CE activities quickly. Ongoing needs assessment of target audiences and evaluation of the effectiveness of Internet CE is essential to optimizing the role of this educational format for healthcare professionals. PMID- 12887231 TI - Nosocomial respiratory syncytial virus infection in a pediatric ward. PMID- 12887232 TI - Three, if by air. PMID- 12887233 TI - Influence of building construction work on Aspergillus infection in a hospital setting. AB - BACKGROUND AND OBJECTIVE: Aspergillus fumigatus is a major pathogen causing nosocomial infections. Hospital outbreaks of invasive aspergillosis have been associated with the renovation and construction of buildings. Building construction work for fire safety upgrading was undertaken during a 16-week period in 2001 at Box Hill Hospital. This study was designed to examine the effect of construction on invasive aspergillosis when using standard and additional protective measures. METHODS: Baseline air sampling was conducted in 18 areas. The validity of the air sampling was assessed by comparing the ability of two air samplers to detect Aspergillus conidia. Surveillance of nosocomial Aspergillus infection was conducted by reviewing the records of patients with a sputum culture positive for Aspergillus and those prescribed amphotericin or itraconazole for the period of construction activity and the same period the previous year. RESULTS: Aspergillus was isolated infrequently and there was no statistically significant difference in the levels of viable pathogenic fungi between areas w here construction work was undertaken and areas where it wasnot undertaken. A moderate agreement was observed between the two air samplers (kappa = 0.4; P < .05). There was no difference in the incidence of invasive aspergillosis between 2000 and 2001 (incidence density ratio, 1.2; 95% confidence interval, 0.3 to 4.1). CONCLUSION: The influence of construction work performed with protective measures needs to be examined in an environment with higher levels of airborne fungi to confirm the findings of this study. PMID- 12887234 TI - Invasive pulmonary aspergillosis soon after therapy with infliximab, a tumor necrosis factor-alpha-neutralizing antibody: a possible healthcare-associated case? AB - BACKGROUND: Infliximab is a chimeric monoclonal antibody against tumor necrosis factor (TNF)-alpha, used for the treatment of Crohn's disease and rheumatoid arthritis. Recently, an increased risk of infection due to Mycobacterium tuberculosis and rare cases of invasive fungal disease have been reported following infliximab therapy. CASE REPORT: A 73-year-old woman with chronic rheumatoid arthritis who had been treated with methotrexate, leflunomide, and prednisone was given the first of three doses of infliximab in June 2001. In July 2001, she presented with cough, and in August, she had a right upper lobe infiltrate that was treated with levofloxacin without improvement. In October, the patient had right upper and middle lobe infiltrates on a chest X-ray and computed tomography scan. At bronchoscopy, an endobronchial mass was biopsied, which demonstrated Aspergills fumigatus. Our patient had frequently accompanied her daughter on visits to another medical center following a stem cell transplant, where her daughter was instructed to wear a mask during all visits because of extensive building construction. We postulate that our patient may have acquired pulmonary aspergillosis during this period. Literature reviews on granulomatous diseases following infliximab therapy and hospital-acquired aspergillosis are presented. CONCLUSION: The temporal relationship between the administration of infliximab and A. fumigatus infection in this patient suggests a causal relationship and possible healthcare-associated acquisition. These data underscore the importance of both patient and family education on prevention strategies when potent immune-modulating medications such as infliximab have been prescribed. PMID- 12887235 TI - Efficacy of environmental measures in reducing potentially infectious bioaerosols during sputum induction. AB - OBJECTIVE: To evaluate the airborne viable bacterial concentrations generated during sputum induction and their reduction with exhaust ventilation, ultraviolet germicidal irradiation (UVGI), or both. METHODS: Exhaust ventilation, upper air UVGI lights, and a portable UVGI unit were operated independently or in combination while and after sputum induction was performed for 58 patients suspected of having active tuberculosis. Viable airborne bacteria were sampled with volumetric air samplers, grown on blood agar, and identified with standard techniques. RESULTS: During and immediately after sputum induction, concentrations of airborne bacteria, particularly respiratory tract or oropharyngeal organisms, increased rapidly, regardless of environmental conditions. The subsequent rate of reduction of airborne bacteria was most rapid with the portable UVGI unit, followed by upper air UVGI with air mixing. Exhaust ventilation achieved high air changes per hour, but efficacy in reducing airborne bacterial concentrations was low. However, the continuous entrainment of bacteria laden air from the hallway outside may have resulted in underestimation. The efficacy of a wall-mounted upper air UVGI fixture was significantly less if there was no air mixing. The irradiation from this fixture was of adequate germicidal intensity only in a narrow horizontal plane 2.5 m above the floor. CONCLUSION: Sputum induction was associated with a rapid and substantial increase in airborne bacteria despite the use of exhaust ventilation providing more than 30 air changes per hour, and the adjunct use of UVGI. This emphasizes that healthcare workers involved in similar cough-inducing procedures performed for patients with suspected tuberculosis must wear appropriate personal respirators. PMID- 12887236 TI - Molecular epidemiology of systemic infection caused by Enterobacter cloacae in a high-risk neonatal intensive care unit. AB - OBJECTIVE: To investigate the molecular epidemiology of systemic nosocomial infections caused by Enterobacter cloacae. SETTING: Neonatal intensive care unit (NICU) of a tertiary-care university hospital. PATIENTS: Forty-two high-risk neonates with systemic infections caused by E. cloacae. METHODS: From 1995 to 1997, the variables associated with death in these patients were evaluated. The molecular epidemiology of the strains responsible for the systemic infections, and 14 unrelated strains, was studied using plasmid analysis and pulsed-field gel electrophoresis (PFGE). RESULTS: The overall mortality rate for infection caused by E. cloacae was 34%, whereas the crude mortality rate during the study period was 8.12% (P < .001). Gestational age (preterm neonates) and birth weight (small for gestational age) were not associated with a higher risk of death. Insertion of a venous catheter by dissection of a peripheral vein was the only invasive procedure related to death (P = .016) in this study. A molecular analysis showed that three outbreaks, each occurring in a different year, were caused by strains with distinctive DNA profiles. Only one outbreak was identified by the infection control service, in the NICU. Plasmid analysis and PFGE showed similar ability to discriminate control strains from the E. cloacae strains isolated from the neonates. CONCLUSIONS: Systemic infections caused by E. cloacae in our NICU were associated with a high mortality rate and occurred as small, unrecognized outbreaks. These results may not be generalizable because the data were from a single center. PMID- 12887237 TI - Analysis of three outbreaks due to Klebsiella species in a neonatal intensive care unit. AB - OBJECTIVE: To investigate the clinical, microbiological, and epidemiologic features of three outbreaks caused by Klesiella during 3 years. SETTING: Neonatal intensive care unit of a university hospital. PATIENTS: Thirty affected neonates. METHODS: Data were collected through chart reviews and conversations with physicians. Screening samples were obtained from the staff, the neonates, and the environment. Antibiogram typing and arbitrarily primed polymerase chain reaction based fingerprinting were used to type the strains. RESULTS: The first outbreak had 13 K. pneumoniae strains isolated. The second outbreak had 10 K. oxytoca strains isolated. The third outbreak had 20 K. pneumoniae strains isolated. More than half of the patients had low birth weights, were premature, and underwent mechanical ventilation and intravenous catheterization. Approximately three fourths of the patients died. The isolates tested were completely susceptible to meropenem, cefoxitin, and ciprofloxacin and were resistant to cephalothin. More than half of these strains were resistant to many beta-lactam antibiotics, amikacin, and trimethoprim/sulfamethoxazole. Typing procedures yielded 3 antibiotypes and 3 genotypes among the isolates of the first outbreak, 3 antibiotypes with 1 subtype and 2 genotypes with 1 subtype in the second outbreak, and 2 antibiotypes and 2 genotypes in the third outbreak. CONCLUSIONS: Klebsiella outbreaks mainly affected premature neonates with intravenous catheters, mechanical ventilation, or both. The high mortality rate (76.7%) was notable. Resistance to multiple antibiotics, but mainly to broad-spectrum beta lactam antibiotics, was observed, particularly in K. pneumoniae isolates. Molecular typing indicated that the three outbreaks were not related to one other. PMID- 12887238 TI - Reducing central venous catheter-associated primary bloodstream infections in intensive care units is possible: data from the German nosocomial infection surveillance system. AB - BACKGROUND AND OBJECTIVE: The German Nosocomial Infection Surveillance System (KISS) began in 1997 as a nationwide surveillance project for voluntary registration of nosocomial infections in intensive care units (ICUs). This study investigates trends in the rates of central venous catheter (CVC)-associated primary bloodstream infections (BSIs) in ICUs since participation in KISS. METHODS: Eighty-four ICUs that had participated in KISS for at least 24 months were considered for more detailed analysis. Monthly rates of primary BSI for the 84 ICUs were pooled for the 24 months. The best model for describing the curve of reduction was sought. Additionally, incidence densities were compared using the z test. RESULTS: For the 212 ICUs participating, a relative 25.7% decrease (from 2.1 to 1.6 primary BSIs per 1,000 CVC-days) was observed from January 1997 to June 2001. The 84 ICUs that participated in KISS for a minimum of 24 months accumulated 552,359 patient-days and 404,897 CVC-days during their 24 months. A linear regression model was selected to explain the curve of primary BSI reduction in the 84 ICUs. It showed a decrease from 2.1 to 1.5 primary BSIs per 1,000 CVC-days, meaning an overall relative reduction of 28.6% during the 2-year observation period. These results were significant (Student's t test for the monthly reduction coefficient; P = .04). The reduction of primary BSIs was shown for both clinical sepsis and laboratory-confirmed, CVC-associated primary BSIs. CONCLUSION: Performing surveillance with KISS was associated with a reduction of the rates of CVC-associated primary BSIs in ICU patients. PMID- 12887239 TI - Evaluation of the antimicrobial efficacy of urinary catheters impregnated with antiseptics in an in vitro urinary tract model. AB - OBJECTIVES: To evaluate the long-term efficacy of urinary Foley catheters (latex and silicone) impregnated with (1) chlorhexidine and silver sulfadiazine (CXS) and (2) chlorhexidine, silver sulfadiazine, and triclosan (CXST) in inhibiting extra-luminal bacterial adherence and to compare their efficacy with that of silver hydrogel latex (SH) and nitrofurazone-treated silicone (NF) catheters. DESIGN: The antimicrobial spectrum of these catheters was evaluated using a zone of inhibition assay. A novel in vitro urinary tract model was developed to study the potential in vivo efficacy of antimicrobial catheters in preventing extraluminal bacterial colonization. The "meatus" was inoculated daily with Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, Enterococcus faecalis, Pseudomonas aerginosa, and Candida albicans. The "bladder" portion of the model was cultured daily to determine bacterial growth. RESULTS: Both CXS and CXST catheters had a broader antimicrobial spectrum than SH and NF catheters. In the in vitro model, CXST latex and silicone catheters exhibited significantly better efficacy (3 to 25days) against uropathogens, compared with CXS (1 to 14 days) and control (0 to 5 days) catheters (P = .01). CXST latex catheters exhibited significantly longer protection against Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, and Pseudomonas aeruginosa, compared with SH catheters (P = .01). CXST silicone catheters resisted colonization with Staphylococcus aureus and Staphylococcus epidermidis for a significantly longer period (23 to 24 days) than did NF catheters (9 to 11 days) (P = .01). CONCLUSION: Catheters impregnated with synergistic combinations of chlorhexidine, silver sulfadiazine, and triclosan exhibited broad-spectrum, long term resistance against microbial colonization on their outer surfaces. PMID- 12887240 TI - An outbreak of epidemic keratoconjunctivtis in a pediatric unit due to adenovirus type 8. AB - OBJECTIVES: To investigate and control an outbreak of epidemic keratoconjunctivitis (EKC) occurring in a neonatal intensive care unit (NICU) and to determine signs specific for EKC in newborns. DESIGN: Outbreak investigation and case-control study. SETTING: NICU of a 1,600-bed university hospital in Heidelberg, Germany. PATIENTS: Case-patients were defined as individuals who had adenoviral antigen detected by ELISA or by PCR from conjunctival swabs or who had clinical signs of conjunctivitis. Twelve newborns from the NICU who had EKC between October and November 1998 were compared with 11 who had bacterial conjunctivitis. INTERVENTION: Control measures included cohorting patients in contact isolation, using gloves and gowns for patient care, and using a hand rub and disinfectants with virucidal activity. RESULTS: Thirteen patients, 6 healthcare and 11workers, relatives of the patients had EKC, according to our case definition. Case-patients were more likely than control-patients to develop lacrimal swelling (P < .001), eye redness (P = .004), and lacrimation (P = .037) involving both eyes (P = .002). Prior examination by an ophthalmologist was a significant risk factor for EKC (P = .004). For diagnosis or treatment of retinopathy, premature newborns were seen by an ophthalmologist from a nearby eye care center where an EKC outbreak was ongong. No new cases were diagnosed more than 10 days after the implementation of control measures. CONCLUSION: In newborns, lacrimal swelling, eye redness, and lacrimation in both eyes should evoke suspicion of EKC. Ophthalmologists who have had contact with known EKC cases should use antiseptics and disinfectants with virucidal activity before contact with newborns or abstain from examining newborns. PMID- 12887241 TI - Reported measures of hygiene and incidence rates for hospital-acquired diarrhea in 31 French pediatric wards: is there any relationship? AB - OBJECTIVE: To investigate the relationship between hygienic measures reported for the prevention of hospital-acquired diarrhea and incidence rates of hospital acquired diarrhea. DESIGN: A survey of hospital-acquired diarrhea was conducted between January 1 and March 31, 1999. Multivariate analysis of reported measures of hygiene according to the observed incidence rates of hospital-acquired diarrhea was performed. SETTING: Thirty-one pediatric or neonatal wards located in hospitals in the southeast of France, selected as a convenience sample of wards volunteering to participate. PATIENTS: A total of 6,726 children younger than 5 years. RESULTS: The overall incidence rate of hospital-acquired diarrhea was 3.6%. Rotavirus was responsible for 69% of the cases of hospital-acquired diarrhea. Among the hygienic measures reported by the wards for preventing hospital-acquired diarrhea were using a single room or cohorting (77.4%), washing hands (83.9%), wearing gowns (80.6%), and wearing disposable single-use gloves for diapering a patient (51.6%). By multivariate analysis, the variables statistically associated with a lower incidence of hospital-acquired diarrhea were restricting the patient's mobility outside his or her room, keeping the patient's door closed, and having fewer than 20 beds in the ward, with adjusted odds ratios of 0.34 (95% confidence interval [CI95], 0.18 to 0.65), 0.33 (CI95, 0.23 to 0.47), and 0.42 (CI95 0.30 to 0.60), respectively. CONCLUSION: Simple preventive measures can decrease the rate of hospital-acquired diarrhea in pediatric wards. PMID- 12887242 TI - Increasing pneumococcal vaccination rates among hospitalized patients. AB - OBJECTIVE: To increase the proportion of inpatients vaccinated against pneumococcal infection. DESIGN: Pre- and post-intervention study. SETTING: University medical center-affiliated, suburban community teaching hospital. PATIENTS: Unvaccinated inpatients 65 years and older and those 2 to 64 years old who had chronic medical conditions predisposing them to invasive pneumococcal infection. INTERVENTION: The nursing staff screened newly admitted patients for eligibility based on age, diagnosis, or medications from a computer-generated admissions list and placed a pre-printed order form for the pneumococcal polysaccharide vaccine (PPV) on the charts of eligible patients. Following the physician's order, the nursing staff administered the PPV and recorded it Ongoing quality improvements including admission vaccination screening and computer-based record keeping were initiated to identify unvaccinated eligible patients and track vaccination status. RESULTS: Efforts resulted in rates of in-hospital vaccination ranging from 3.1% to 7.9% (mean, 5.2% +/- 1.7% [standard deviation]) and significant improvements in the assessment of previous vaccination status, reaching 54% of eligible patients after 1 year. Ascertainment of a previous vaccination increased significantly following the initiation of the use of admission forms that specifically assessed vaccination status and a system to permanently record vaccination status in an electronic medical record (P < .05). CONCLUSION: Concerted efforts using electronic medical records significantly improved the assessment and documentation of inpatient vaccination status. Greater improvement of the rates of in-hospital vaccination will require healthcare system-wide efforts such as a standing order policy for vaccinating all eligible patients. Standing orders for inpatient immunization supported by effective assessment and tracking systems have the potential to raise vaccination rates to the goals of Healthy People 2010. PMID- 12887243 TI - Multi-society guideline for reprocessing flexible gastrointestinal endoscopes. Society for Healthcare Epidemiology of America. AB - Flexible gastrointestinal endoscopy is a valuable diagnostic and therapeutic tool for the care of patients with gastrointestinal and pancreaticobiliary disorders. Compliance with accepted guidelines for the reprocessing of gastrointestinal endoscopes between patients is critical to the safety and success of their use. When these guidelines are followed, pathogen transmission can be effectively prevented. Increased efforts and resources should be directed to improve compliance with these guidelines. Further research in the area of gastrointestinal endoscope reprocessing should be encouraged. The organizations that endorsed this guideline are committed to assisting the FDA and manufacturers in addressing critical infection control issues in gastrointestinal device reprocessing. PMID- 12887244 TI - Management of varicella-vaccinated patients and employees exposed to varicella in the healthcare setting. PMID- 12887245 TI - Take two orthogonals and call me in the morning. AB - Analysis of variance (ANOVA) is used to prevent inflated type I error when hypothesis testing involves comparing more than two groups. If an ANOVA result indicates a statistically significant difference exists somewhere within, the next task is to discover exactly which combination or combinations of those groups account for the significant difference. Among many methods available for that exploration, orthogonal contrasts and relatively simple graphs are noteworthy. PMID- 12887246 TI - Respiratory syncytial virus pneumonia in an adult renal transplant patient: an unexpected nosocomial infection. AB - This article describes a case of respiratory syncytial virus pneumonia believed to have been acquired nosocomially in an adult renal transplant recipient. The mode of transmission, diagnosis, and prevention of infection due to this virus in the immunocompromised patient are discussed. PMID- 12887247 TI - The scientific process: new forces attempt to enter the scene. PMID- 12887248 TI - Time-course and levels of apoptosis in various tissues of black tiger shrimp Penaeus monodon infected with white-spot syndrome virus. AB - This study focused on apoptosis in various tissues of the black tiger shrimp Penaeus monodon following white spot syndrome virus (WSSV) injection. The study included: (1) light microscopy (LM) and transmission electron microscopy (TEM) of various tissues; (2) fluorescent LM of nuclear DNA by staining with 4, 6 diamidine-2-phenyl indole dihydrochloride (DAPI) and TdT-mediated dUTP nick-end labelling (TUNEL) techniques; and (3) determination of caspase-3 activity. Juvenile P. monodon were injected with WSSV, and several tissues of ectodermal and mesodermal origin were studied at different intervals after injection. The total haemocyte count had decreased to one-tenth of its original level 60 h after WSSV injection. By LM, extensive destruction by WSSV was observed in the stomach epithelium, gills, hematopoietic tissue, hemocytes and the heart, but the most severely affected tissue was the subcuticular epithelium. TEM revealed that at 6 h post-injection (p.i.) the chromatin of infected nuclei was marginated, and by 24 h p.i. the nuclei were filled with enveloped and non-enveloped WSSV virions. At later stages of the infection, the nucleus extruded WSSV particles. Chromatin margination and nuclear condensation and fragmentation (i.e. signs of apoptosis) were observed as early as 6 h p.i. in all affected tissues, but occurred in cells without WSSV virions rather than in cells with virions. The occurrence of apoptosis was supported by data obtained using TUNEL and by DAPI-staining and progressed from 6 to 60 h p.i. In addition, caspase-3 activity in WSSV-infected shrimp was about 6-fold higher than that in uninfected shrimp. The data strongly suggests that apoptosis occurs following WSSV infection in P. monodon, but the extent to which it contributes to shrimp mortality requires further investigation. PMID- 12887249 TI - Molecular characterisation of Neoparamoeba strains isolated from gills of Scophthalmus maximus. AB - Small subunit ribosomal RNA gene sequences were determined for 5 amoeba strains of the genus Neoparamoeba Page, 1987 that were isolated from gills of Scophthalmus maximus (Linnaeus, 1758). Phylogenetic analyses revealed that 2 of 5 morphologically indistinguishable strains clustered with 6 strains identified previously as N. pemaquidensis (Page, 1970). Three strains branched as a clade separated from N. pemaquidenis and N. aestuarina (Page, 1970) clades. Our analyses suggest that these 3 strains could be representatives of an independent species. In a more comprehensive eukaryotic tree, strains belonging to Neoparamoeba spp. formed a monophyletic group with a sister-group relationship to Vannella anglica Page, 1980. They did not cluster with Gymnamoebae of the families Hartmannellidae, Flabellulidae, Leptomyxidae or Amoebidae presently available in GenBank. PMID- 12887250 TI - Efficacy of selected oral chemotherapeutants against Ichthyophthirius multifiliis (Ciliophora: Ophyroglenidae) infecting rainbow trout Oncorhynchus mykiss. AB - The chemotherapeutic efficacy of 6 in-feed compounds against Ichthyophthirius multifiliis Fouquet, 1876 was assessed using experimental infections of rainbow trout Oncorhynchus mykiss (Walbaum) fingerlings. Trial doses of 104 ppm amprolium hydrochloride or 65 ppm clopidol fed to fish for 10 d prior to infection significantly reduced the number of trophonts establishing in trout fingerlings by 62.0 and 35.2% respectively. In-feed treatments of infected trout with either 63 or 75 ppm amprolium hydrochloride, 92 ppm clopidol, or 38, 43 or 47 ppm salinomycin sodium for 10 d also significantly reduced the number of surviving trophonts by 77.6 and 32.2% for amprolium, 20.1% for clopidol and 80.2, 71.9 and 93.3% respectively for salinomycin sodium. PMID- 12887251 TI - Immunization of rainbow trout Oncorhynchus mykiss against Discocotyle sagiffata (Monogenea). AB - Rainbow trout Oncorhynchus mykiss were injected intraperitoneally with 2 different Discocotyle sagittata extracts dissolved in PBS and subsequently exposed to controlled infection. Immunization resulted in significantly reduced (p < 0.0001) worm intensities in > 50% of vaccinated fish (response arbitrarily defined as parasite burdens < mean control intensity - 1 SD), irrespective of the immunization regime (different parasite extracts, dosing and application schedules) employed. The protective effect of worm extract applied in Freund's complete adjuvant (FCA) did not differ significantly from extract given in PBS. Vaccination with embryonated parasite eggs extract and with FCA alone did not result in partial immunity, suggesting the observed protective effect is specific. Immunized fish had significantly higher specific antibody titres at the time of dissection (as determined by ELISA) than both naive and control fish. Overall, a significant negative correlation was found between antibody titres and worm burdens, suggesting immunoglobulins are implicated in mediating partial immunity. Western blot tests indicated the 2 different worm extracts used to immunize fish share antigens, but each one primarily induced recognition of a distinct band (30 and 38 kDa). Immunization seems to promote a shift between 2 equilibria, rather than progressively increasing protection. This would explain why boosting did not increase immunity, and why 2 different extracts primarily inducing recognition of 2 distinct antigens provide similar degrees of protection. Although several other non-specific and cellular factors are likely to be involved in controlling parasite numbers, it cannot be excluded that antibodies could be involved in mediating the observed partial immunity. PMID- 12887252 TI - Evaluation of a vector-control strategy of haemorrhagic thelohanellosis in carp, caused by Thelohanellus hovorkai (Myxozoa). AB - The life cycle of Thelohanellus hovorkai (Myxozoa), the causative agent of haemorrhagic thelohanellosis of carp Cyprinus carpio, involves the alternate oligochaete host Branchiura sowerbyi, which plays the role of vector in the parasite's transmission. Field investigations in carp farms suggested that oligochaete fauna were closely associated with the substrate type of the pond. The pond bottom in the enzootic farm consisted of clay soil and soft sediments comprised of organic mud, in which B. sowerbyi dominated in high densities, with a maximum of 5.6 ind. kg(-1) soil. In another case, in a carp farm with no previous history of the disease, the pond bottom was sandy soil, in which small sized oligochaetes, composed mainly of Limnodrilus socialis, dominated. Laboratory studies on the substrate preference of oligochaetes proved that B. sowerbyi prefers mud to sand, whereas L. socialis has no tendency to substrate tropism. The delicate body surface of B. sowerbyi was subject to damage by rugged edged sand particles, which inflicted severe injuries to the worms. Transmission experiments showed that L. socialis, which are non-susceptible to T. hovorkai, suppressed the production of T. hovorkai actinospores in B. sowerbyi in a mixed assemblage of oligochaetes. Field and experimental evidence in this study imply that substrate replacement in culture ponds might regulate the benthic oligochaete communities, resulting in minimization of the impact of haemorrhagic thelohanellosis. We propose that ecological control of oligochaete fauna by environmental management is a promising strategy against myxozoan diseases. PMID- 12887253 TI - Susceptibility of two strains of rainbow trout (one with suspected resistance to whirling disease) to Myxobolus cerebralis infection. AB - The susceptibility of 2 strains of rainbow trout Oncorhynchus mykiss, 1 from North America (TL) and 1 from Germany (GR), to Myxobolus cerebralis (the cause of salmonid whirling disease) was assessed following exposure to the infectious stages (triactinomyxons). Two laboratory experiments were conducted with age matched rainbow trout of each strain. At the beginning of the study, the 2 trout strains were aged ca. 570 degree-days in Expt 1, and ca. 999 degree-days in Expt 2. In both experiments, replicate groups of each trout strain were exposed to 10, 100, 1000 or 10000 triactinomyxons (TAMs) fish(-1) for 2 h. The fish were then held in aquaria receiving 15 degrees C well-water. Severity of infection was evaluated 5 mo after exposure by presence of clinical signs (whirling and/or black tail), prevalence of infection, severity of microscopic lesions, and spore counts. Clinical signs of whirling disease were evident only in the younger fish exposed in Expt 1: These occurred first among TL rainbow trout at the highest dose at 6 to 7 wk post exposure and then 2 wk later in fish at the 1000 TAMs dose. Black tail was also observed among GR rainbow trout at the 10000 TAMs dose only, but in fewer fish. The prevalence of infection, spore numbers, and severity of microscopic lesions due to M. cerebralis among GR rainbow trout were less at all doses compared to TL rainbow trout. Risk of infection analyses showed that TL rainbow trout were more prone to infection at the lower doses than GR trout. Mean spore counts were consistently (10- to 100-fold) less in GR than TL trout at doses of 1000 TAMs or lower. Microscopic lesions increased with increasing dose in both strains of rainbow trout. The mechanisms underlying the greater resistance of the GR strain to M. cerebralis infections are unknown, but are under investigation as part of a long-term project to determine the basis for resistance and susceptibility of salmonid fishes to whirling disease. PMID- 12887254 TI - Characterisation of two novel types of hexactinomyxon spores (Myxozoa) with subsidiary protrusions on their caudal processes. AB - Two types of hexactinomyxon spores, Hexactinomyxon type 1 nov. and Hexactinomyxon type 2 nov., are reported from freshwater tubificid oligochaetes, Limnodrilus hoffmeisteri and L. udekemianus. Spores are triradially symmetrical and comprise a spore body, style and 6 caudal processes. The caudal processes arise from the division of each of the 3 valve cells into an equal pair of projections at the base of the style. One of each pair is fused conspicuously to its nearest neighbour for the initial 1/5 to 1/4 of their total length. Distally, each process possesses subsidiary protrusions which are irregularly distributed and irregularly shaped extensions of the valve cell. Scanning electron microscopy of Hexactinomyxon type 2 nov. revealed that these protrusions are a seamless extension of the valve cell wall which branch distally, occasionally laterally, and terminate in a distinct bulbous structure; they also form the terminus of each process. The small subunit ribosomal DNA gene (18S) of both hexactinomyxon types was amplified through a nested PCR, then digested with the restriction enzymes Dde I and Hha I. The resultant cleavage patterns suggested the presence of 2 forms. Subsequent partial sequencing of 18S rDNA confirmed the identification of 2 novel types. PMID- 12887255 TI - Light- and electron-microscope description of Kudoa paralichthys n. sp. (Myxozoa, Myxosporea) from the brain of cultured olive flounder Paralichthys olivaceus in Korea. AB - A new Myxosporea, Kudoa paralichthys n. sp., is described from the brain of cultured olive flounder Paralichthys olivaceus in South Korea. Mature spores were quadrate in apical view, measuring 5.19 +/- 0.54 microm in length, 8.23 +/- 0.50 microm in width, and 6.87 +/- 0.45 microm in thickness. Four valves were equal in size, each with 1 polar capsule. Polar capsules were pyriform in shape, measuring 2.2 +/- 0.22 microm in length and 1.2 +/- 0.14 microm in breadth. The sporoplasm consisted of a larger outer cell completely surrounding a smaller inner one, and had cytoplasmic projections. The junctions of shell valves were L-shaped. The sutural planes converged at the anterior ends of the spores and were associated with 4 small apex prominences in the central meeting point of the spores. PMID- 12887256 TI - Emerging disease of amphibians cured by elevated body temperature. AB - The emerging infectious disease chytridiomycosis is thought to have contributed to many of the recent alarming declines in amphibian populations. Mortalities associated with these declines have often occurred during cooler seasons and at high elevations, suggesting that environmental temperature may be an important factor in disease emergence. We found that thermal environment affects the progress of the disease, and that housing frogs Litoria chloris at an environmental temperature of 37 degrees C for less than 16 h can clear them of the chytrid pathogen Batrachochytrium dendrobatidis. Our experiment demonstrated that elevated body temperatures similar to those experienced in behavioral fever and during normal thermoregulation can clear frogs of chytrid infection; therefore, variation in thermoregulatory opportunities and behaviors are likely to contribute to the differences in disease incidence observed among host species, populations, and regions. Although further refinement of the technique is needed to encompass various host species, appropriately applied thermal manipulations of amphibians and their enclosures may prove to be a safe and effective way of eliminating the fungal pathogen from captive amphibian populations and: preventing accidental spread of the pathogen when animals are translocated or released from captivity. PMID- 12887257 TI - Fecal samples as DNA source for the diagnosis of Necrotizing Hepatopancreatitis (NHP) in Penaeus vannamei broodstock. AB - Necrotizing Hepatopancreatitis (NHP) is a severe disease of cultivated penaeid shrimp caused by a pleomorphic, gram-negative, intracellular rickettsia-like bacterium. Current diagnostic methods for this disease are invasive, requiring dissection of the animal to perform histopathological analysis. In Colombia, NHP affects mainly broodstock, being a major cause of mortalities in maturation laboratories. In order to identify the presence of NHP without having to dissect the animal, we developed a PCR-based method using fecal samples as the DNA source. The DNA was extracted using a quick isolation protocol followed by amplification with primers specific for 16S ribosomal RNA gene sequences. To verify the sensitivity and specificity we analyzed samples from the same animal by PCR and in situ hybridization, and found 100% agreement. In addition, we amplified DNA extracted form paraffin blocks to confirm NHP diagnosis. PCR amplification from fecal samples and paraffin blocks yielded the expected 440 bp fragment. We conclude that PCR amplification from fecal samples is a valuable tool for the diagnosis of NHP in broodstock organisms, and that paraffin-fixed tissues can be used as a source of DNA for PCR amplification of NHP. PMID- 12887258 TI - Gastrointestinal helminths of Risso's dolphin Grampus griseus from the Western Mediterranean. AB - The gastrointestinal helminth fauna of 17 Risso's dolphins Grampus griseus (Cuvier, 1812) from the Western Mediterranean was analysed. Five helminth species were found. Adults and juveniles of Hadwenius delamurei were collected from the duodenum of 7 animals. Adults were located significantly more anteriorly than juveniles, which is compatible with a previous hypothesis that species of Hadwenius undergo an ontogenetic migration towards the anterior duodenum. Eggs from Pholeter gastrophilus appeared in 1 small cyst in the main stomach. Adults of Tetrabothrius forsteri and Trigonocotyle globicephalae were collected from the duodenum of 1 and 4 individuals, respectively. Metacestodes, Scolex pleuronectis, were found in the liver, pancreas, pyloric stomach and intestine of 13 individuals. The most favored locations were in the mucosa of the terminal colon and in anal crypts. It is likely that worms select these locations to reach the mesenteries and blubber of the anogenital area, where S. pleuronectis might become Monorygma and/or Phyllobothrium. Two individuals of Stenurus globicephalae occurred in the intestine of 1 individual, and were probably swallowed accidentally. The intestinal helminth communities of Risso's dolphin are depauperate. The parasite species found also occur in other oceanic cetaceans from the Western Mediterranean. However, H. delamurei and T. globicephalae are only shared with pilot whales Globicephala spp., which are phylogenetically closely related to Risso's dolphins. PMID- 12887259 TI - Immunofluorescence of the epizootic ulcerative syndrome pathogen, Aphanomyces invadans, using a monoclonal antibody. AB - A monoclonal antibody (MAb), designated 3gJC9, was raised against a protein antigen of Aphanomyces invadans, the oomycete pathogen that causes epizootic ulcerative syndrome (EUS). The antigen was expressed on the surface of hyphae and secreted extracellularly. MAb 3gJC9 did not cross-react with other oomycete or fungal pathogens of fish, although it did react to the crayfish plague pathogen A. astaci. The MAb was used for immunofluorescent staining on histological sections of fish infected with EUS, and was found to be more sensitive than conventional staining methods for detecting A. invadans. It thus has utility in confirming the case definition of EUS. It also revealed very small filamentous structures, the significance of which is unclear, but they may represent an early stage of infection, thus allowing earlier detection of the disease, since they are not detected using conventional staining methods. PMID- 12887260 TI - Early intervention in multiple sclerosis : better outcomes for patients and society? AB - Multiple sclerosis (MS) is thought to be a chronic inflammatory disorder of the CNS. The past decade has seen the introduction of the new immunomodulatory drugs, interferon (IFN)-beta and glatiramer acetate, that have considerably improved the therapeutic options for this often disabling disease. The efficacy of these treatments in terms of reducing relapse rate and slowing progression has been proven in several large, multicentre, randomised, controlled trials. Similarly, early IFNbeta treatment of patients with clinically isolated syndromes suggestive of MS has been shown to lengthen time to conversion into definite MS. Cost effectiveness has been questioned with the increasing use of these innovative and, therefore, costly therapies; however, modern studies with appropriate economic modelling suggest that treatment with IFNbeta may indeed be cost effective. Since increasing disability is associated with increasing costs, stabilisation of the disease at low functional grades of disability should aim at not only improving quality of life for the individual patient, but provide for prospective cost-benefit analysis focussing on the socioeconomic aspects of MS. PMID- 12887261 TI - Calcineurin inhibitors in renal transplantation: what is the best option? AB - Recently, new calcineurin inhibitors, such as tacrolimus (FK-506) and microemulsion cyclosporin, have been approved for maintenance immunosuppression in renal transplant recipients and short-term outcomes have been accumulating. In the majority of patients, these calcineurin inhibitors have been used in combination with new immunosuppressive drugs, such as mycophenolate mofetil (MMF) or sirolimus. Under these circumstances, a comparison of cyclosporin and tacrolimus provides the answer to a very important controversial issue. Which drug should we choose in individual patients? In an attempt to answer this question, this review compared the use of tacrolimus and cyclosporin in modern immunosuppressive regimens, which have already been published in well designed clinical studies, and discusses how immunosuppression should be individualised in renal transplant patients.Overall, short-term patient and graft survival with cyclosporin microemulsion and tacrolimus is almost identical. The incidence of acute rejection is generally lower in tacrolimus/azathioprine- than in cyclosporin/azathioprine-treated patients. However, in conjunction with MMF, the difference in the incidence of acute rejection between tacrolimus- and cyclosporin-treated patients became smaller. Adverse events, such as hypertension, hyperlipidaemia and cosmetic changes (gum hypertrophy, hirsutism) seem to be less frequent in tacrolimus-treated than in cyclosporin-treated patients. Recent randomised studies showed that the incidence of post-transplant diabetes mellitus was almost identical between low-dose tacrolimus- and cyclosporin-treated patients. According to the data discussed in this review, the recommendation on the choice of calcineurin inhibitors at this moment is that either cyclosporin or tacrolimus can be used safely and effectively for patients without any risk factors. However, at our centre, we prefer tacrolimus to cyclosporin in patients with a high risk for rejection, such as those with ABO incompatibility, delayed graft function, sensitisation, and African American race and some other risk factors, such as hypertension and hyperlipidaemia. Moreover, tacrolimus may be preferable to cyclosporin for women because of hirsutism and for children because of the steroid-sparing effect. We consider that cyclosporin should be chosen when patients experience tacrolimus-related adverse events, such as severe chest pain, tremor, gastrointestinal symptoms and encephalopathy. In conclusion, well tolerated and effective immunosuppression is feasible with both cyclosporin and tacrolimus. In the current immunosuppressive regimens, a calcineurin inhibitor, either tacrolimus or cyclosporin, is the essential basic standard immunosuppressant. Clinicians need to decide the best means of optimising therapy for individual patients, based on various risk factors, such as risk of rejection, i.e. sensitisation, delayed graft function and ABO incompatibility, and some adverse events, such as hypertension, hyperlipidaemia and cosmetic changes. PMID- 12887262 TI - Neurotoxic complications of chemotherapy in patients with cancer: clinical signs and optimal management. AB - Neurotoxic side effects of chemotherapy occur frequently and are often a reason to limit the dose of chemotherapy. Since bone marrow toxicity, as the major limiting factor in most chemotherapeutic regimens, can be overcome with growth factors or bone marrow transplantation, the use of higher doses of chemotherapy is possible, which increases the risk of neurotoxicity. Chemotherapy may cause both peripheral neurotoxicity, consisting mainly of a peripheral neuropathy, and central neurotoxicity, ranging from minor cognitive deficits to encephalopathy with dementia or even coma. In this article we describe the neurological adverse effects of the most commonly used chemotherapeutic agents. The vinca-alkaloids, cisplatin and the taxanes are amongst the most important drugs inducing peripheral neurotoxicity. These drugs are widely used for various malignancies such as ovarian and breast cancer, and haematological cancers. Chemotherapy induced neuropathy is clearly related to cumulative dose or dose-intensities. Patients who already have neuropathic symptoms due to diabetes mellitus, hereditary neuropathies or earlier treatment with neurotoxic chemotherapy are thought to be more vulnerable for the development of chemotherapy-induced peripheral neuropathy. Methotrexate, cytarabine (cytosine arabinoside) and ifosfamide are primarily known for their central neurotoxic side effects. Central neurotoxicity ranges from acute toxicity such as aseptic meningitis, to delayed toxicities comprising cognitive deficits, hemiparesis, aphasia and progressive dementia. Risk factors are high doses, frequent administration and radiotherapy preceding methotrexate chemotherapy, which appears to be more neurotoxic than methotrexate as single modality. Data on management and neuroprotective agents are discussed. Management mainly consists of cumulative dose-reduction or lower dose-intensities, especially in patients who are at higher risk to develop neurotoxic side effects. None of the neuroprotective agents described in this article can be recommended for standard use in daily practise at this moment, and further studies are needed to confirm some of the beneficial effects described. PMID- 12887263 TI - Long-term complications of chemotherapy for germ cell tumours. AB - Testicular cancer is the most common solid tumour among young males aged 15-35 years. Cisplatin-based combination chemotherapy has changed the outlook of this disease. Disseminated testicular cancer, once uniformly fatal, now has a cure rate of more than 80% with combination chemotherapy. Systematic randomised trials have shown that cisplatin, etoposide and bleomycin (PEB) combination chemotherapy remains the mainstay of treatment. While there is a high cure rate with chemotherapy in patients with this disease, some long-term complications from chemotherapy have now been recognised, including secondary leukaemia, therapy related solid tumours, nephrotoxicity, neurotoxicity, pulmonary toxicity, vascular toxicity and infertility. Etoposide, a DNA topoisomerase II inhibitor, is a significant risk factor for developing leukaemia; the risk appears to be correlated with the total dose given. Patients receiving cisplatin-based combination chemotherapy for testicular cancer also appear to have a higher relative risk for developing second non-germ cell malignancies; the greatest risks for therapy-related solid tumours were seen with a combination of radiation therapy plus chemotherapy. Long-term vascular toxicities associated with chemotherapy include Raynaud's phenomenon, acute myocardial infarction and cerebrovascular events. Bleomycin is thought to be the most important drug in the pathogenesis of Raynaud's phenomenon, while cisplatin is the most likely agent involved in myocardial infarction. Peripheral neuropathy is the most common form of neurotoxicity observed with cisplatin-based chemotherapy. Risk factors for the development of neural damage include a high cumulative dose of cisplatin, the use of vinblastine and the concomitant development of Raynaud's phenomenon. Cisplatin is also well known to cause significant nephrotoxicity. Approximately 25% of patients present with azoospermia after undergoing combination chemotherapy with a follow up of 2-5 years. Physician awareness of complications associated with chemotherapy is vital to maximise efficacy, minimise toxicity, and preserve quality of life after treatment. Sperm cryopreservation should be considered for patients who desire children. Close monitoring during therapy allows for the early diagnosis of complications, and close follow up of patients after the completion of therapy is necessary to monitor for relapse and development of long term complications such as myelodysplastic syndrome and leukaemia. Despite these complications, given the potential for cure rates in this young group of patients, the benefits far outweigh the risks. PMID- 12887265 TI - Tenofovir disoproxil fumarate. AB - Tenofovir disoproxil fumarate (tenofovir DF) is a prodrug of tenofovir, a nucleotide reverse transcriptase inhibitor. In two large, well designed, placebo controlled clinical trials, tenofovir DF 300 mg/day resulted in significant reductions in HIV-1 RNA from baseline compared with placebo at 24 weeks in antiretroviral-experienced patients with HIV infection. Patients in both treatment groups continued to receive existing stable antiretroviral therapy. In an extension phase of one trial, these reductions in viral load were maintained after 96 weeks of treatment with tenofovir DF. Preliminary data from a large, 3 year comparative trial suggest the clinical efficacy of tenofovir DF in combination with baseline antiretroviral therapy is similar to that of stavudine in antiretroviral-naive patients with HIV infection. Virological substudies showed that viral suppression was maintained in patients who developed new reverse transcriptase mutations during tenofovir DF therapy (in combination with existing stable antiretroviral drugs) for up to 48 weeks. Isolates of HIV infrequently developed the K65R mutation during 96 weeks of tenofovir DF therapy. Tenofovir DF is generally well tolerated. The most commonly observed adverse events seen with tenofovir DF (in combination with other antiretroviral drugs) were predominantly of a gastrointestinal nature. PMID- 12887264 TI - Current concepts of the pathogenesis of acne: implications for drug treatment. AB - The pathogenesis of acne is complex, with strong evidence supporting the involvement of sebaceous hyperplasia, follicular hyperkeratinisation, bacterial hypercolonisation, as well as immune reactions and inflammation. High sebum concentrations and follicular hyperkeratinisation lead to a change of the follicular milieu with consecutive proliferation of bacteria, chiefly Propionibacterium acnes. This leads to further increased production of the pro inflammatory cytokines interleukin-1alpha and tumour necrosis factor alpha by T cells and keratinocytes, leading to proliferation of both cell types. Follicular keratinocytes fail to differentiate by apoptosis and produce hypergranulosis similar to the impermeable skin outer layer, resulting in the formation of microcomedones. Further inflammatory responses lead to the development of increasing degrees of severity in inflammatory forms of acne. Retinoids aid the differentiation and reduce the hyperproliferation of keratinocytes, and can inhibit the migration of leucocytes. Combination therapy using retinoids plus benzoyl peroxide or antibacterials can treat existing acne lesions faster than the individual agents alone and can also prevent the development of new lesions. The new retinoids (e.g. adapalene) have not only the typical potent comedolytic activity but also anti-inflammatory effects. When added to antibacterial therapy, topical retinoids demonstrate faster and significantly greater reduction of inflammatory acne lesions and comedones than antibacterials alone. PMID- 12887266 TI - Tenofovir disoproxil fumarate. PMID- 12887267 TI - Tenofovir disoproxil fumarate. PMID- 12887268 TI - Tipranavir. AB - Tipranavir is a potent and selective non-peptidic HIV-1 protease inhibitor with a markedly improved resistance profile compared with traditional, peptidomimetic protease inhibitors. The presence of five or fewer protease gene mutations or one or two protease inhibitor resistance-associated mutations (PRAMs) is associated with reduced susceptibility to currently available protease inhibitors. However, 16-20 mutations (including three or more PRAMs) may be needed to confer resistance to tipranavir. Tipranavir-based therapy achieved sustained viral suppression for more than 48 weeks in a small phase II trial in multiple protease inhibitor-experienced HIV-infected patients. A large dose-finding study demonstrated potent virological reduction through 14 days of functional monotherapy in heavily pretreated HIV-infected patients with 6 to >20 protease gene mutations at baseline. Two large, ongoing, phase III trials in patients with multi-drug resistant HIV infection are comparing the efficacy of tipranavir/ritonavir 500/200mg twice daily plus a patient-individualised background antiretroviral regimen versus other ritonavir-boosted protease inhibitor regimens. In general, tipranavir has been well tolerated in clinical trials. As with other protease inhibitors, the most common adverse events with tipranavir have been gastrointestinal disturbances. PMID- 12887269 TI - Tipranavir. PMID- 12887270 TI - Tipranavir. PMID- 12887271 TI - Regulation of ovarian function by the TGF-beta superfamily and follistatin. AB - The role of follistatin as an activin-binding protein has dominated the study of this molecule for the last 10 years. However, there is emerging evidence that follistatin has a role in modulating the biology of other members of the transforming growth factor beta (TGF-beta) superfamily. This review summarizes the current concepts encompassing follistatin biochemistry as well as molecules with which it is functionally associated. Moreover, the importance of the two follistatin isoforms (follistatin-288 and follistatin-315) is discussed with particular emphasis on the regulation of the ovary. In addition to activin, this review discusses the functions of other members of the TGF-beta superfamily, for example growth differentiation factor 9 (GDF-9), bone morphogenetic protein 15 (BMP-15), BMP-6, BMP-4 and BMP-7, in the ovary, and the potential interactions between follistatin and these growth factors. The complex network of TGF-beta superfamily growth factor members involved in the modulation of ovarian function and the interactions of follistatin with these proteins is highlighted. PMID- 12887273 TI - The 'GO' system--a novel method of microculture for in vitro development of mouse zygotes to the blastocyst stage. AB - A novel system of in vitro culture termed the 'glass oviduct' or 'GO' culture system is described. Mouse zygotes were cultured in pairs to the blastocyst stage in open-ended 1 microl glass capillaries. 'GO' culture supported the development of significantly more hatching or hatched blastocysts than did a standard microdroplet (10 zygotes per 20 microl) control culture (48.3 versus 3.3%, respectively). 'GO' bslastocysts contained significantly larger populations of cells (92+/-3 versus 75+/-3), and inner cell mass (25+/-1 versus 21+/-1) and trophectoderm (68+/-2 versus 53+/-3) subpopulations, compared with microdroplet derived blastocysts. Before blastulation, 'GO'-derived morulae were found to contain significantly more cells than microdroplet-derived morulae (27+/-0.7 versus 14+/-0.5). After implantation, 'GO' blastocysts formed fetuses at a similar rate to microdroplet-derived blastocysts (55 versus 62%), but at a lower rate than blastocysts derived in vivo (80%). 'GO'- and microdroplet-derived fetuses were similar in wet weight to each other (0.412 and 0.415 g, respectively) but were heavier than fetuses derived from flushed blastocysts (0.390 g). An additional experiment investigated whether the beneficial effect of 'GO' culture was due to the significantly increased embryo density. Proportions of hatching or hatched blastocysts after 'GO' culture (50%) were higher than after standard microdroplet culture (7.6%), but were not different from culture in high embryo density microdroplets (20 zygotes per 10 microl; 42%). 'GO' blastocysts contained more cells (79.6+/-2.1) than did standard microdroplet derived blastocysts (68.7+/-2.0), but were similar to high density microdroplet derived blastocysts (85.8+/-2.7). Similarly, 'GO' blastocysts contained more trophectoderm cells (62.2+/-2.0) than did standard microdroplet-derived blastocysts (52.7+/-1.7), but were similar to the high density microdroplet blastocysts (68.8+/-2.5). Numbers of inner cell mass cells ('GO', standard microdroplet and high density microdroplet culture) were not different from each other (17.4+/-0.5, 16+/-0.5 and 17+/-0.4, respectively). In conclusion, the 'GO' culture system represents an alternative method to the microdroplet system for small numbers of preimplantation embryos, without detriment to implantation potential. PMID- 12887272 TI - Uterine natural killer cells: insights into their cellular and molecular biology from mouse modelling. AB - In primates, including women, and in rodents, natural killer lymphocytes (NK cells) have a unique relationship with the decidualizing uterus. Implantation sites from genetically modified and transplanted mice have proven useful models for understanding potential mechanisms involved in the recruitment, activation and functions of human CD56(bright) uterine (u)NK cells. Key findings are reviewed in this article. In mice, uNK precursor cells are recruited from secondary lymphoid tissues and are activated coincident with their uterine arrival. uNK cells proliferate, produce cytokines (interferon gamma (IFN-gamma) and interleukin 18 (IL-18) and IL-27), and terminally differentiate into granulated lymphocytes. Many uNK cells proliferate within the myometrium at each implantation site forming a structure, the mesometrial lymphoid aggregate of pregnancy (MLAp) that surrounds blood vessels servicing each placenta. Post mitotic uNK cells are abundant within decidua basalis; frequently (<25%) associating with spiral arteries, intramurally and intraluminally. From mid gestation, numbers of uNK cells decline. Studies of implantation sites in mice lacking uNK cells, IFN-gamma, components of IFN-gamma-induction and -signalling pathways or IFN-gamma-regulated genes indicate that uNK cell-derived IFN-gamma is essential in triggering pregnancy-induced spiral artery modification. Decidual maintenance and uNK cell death are additional effects of uNK cell-derived IFN gamma. Thus, during the first half of gestation, uNK cells contribute to and sustain important changes in the maternal placental bed. PMID- 12887274 TI - Global gene profiling analysis of mouse uterus during the oestrous cycle. AB - Many genes related to the cyclic changes of the uterus during the oestrous cycle have been identified using a one-by-one approach. In the present study, cDNA microarray technology was applied to investigate the global profile of gene expression of mouse uterus at the oestrous and dioestrous stages. At a certain stage of the oestrous cycle, the uteri of mature CD-1 mice (n=10) were removed, pooled and snap-frozen in liquid nitrogen. Total RNA was extracted to synthesize cDNA probes for microarray assay. By screening 8192 mouse genes and expressed sequence tags (ESTs), 51 upregulated and 51 downregulated genes were identified in oestrous uterus, of which 62 are well characterized and 40 are ESTs. The known genes were assigned to various gene categories according to their main function. The microarray was performed three times with three independent sets of uterine tissue pools. The results of northern blot analysis for small proline-rich protein 2 (Sprr2), 17beta-hydroxysteroid dehydrogenase type 2 (17betaHSD-2), high mobility group 2 (Hmg2), mitotic checkpoint component 2 (Mad2) and an EST AW555366 mRNA were consistent with that of microarray analysis. In situ hybridization was performed to localize the transcript of the EST AW555366. Most of the upregulated genes encode secreted immune-related proteins, proteinases and their inhibitors, indicating their potential involvement in sperm viability as well as capacitation. The downregulated genes mainly encode cell cycle-related factors, implying the active proliferation of uterus at dioestrus. PMID- 12887275 TI - Involvement of tyrosine kinase and cAMP-dependent kinase cross-talk in the regulation of human sperm motility. AB - Tyrosine phosphorylation and its upregulation by cAMP have been associated with capacitation and motility changes of spermatozoa. In the present study, washed spermatozoa were incubated for 6 h in protein-supplemented complete medium with or without kinase inhibitors to verify whether upstream activation of protein kinase A is indispensable for tyrosine phosphorylation and motility changes to occur in capacitating human spermatozoa. H89, a specific protein kinase A inhibitor, significantly inhibited the activity of sperm protein kinase A. However, this inhibition did not alter capacitation-related tyrosine kinase activation. Tyrosine phosphorylated proteins, motion parameters and the incidence of phosphotyrosine-immunoreactive spermatozoa were decreased only slightly. Conversely, genistein, a tyrosine kinase inhibitor which inhibited sperm tyrosine kinase but not protein kinase A, significantly reduced all the parameters studied. Spermatozoa incubated with cAMP and pentoxifylline showed a rapid enhancement of tyrosine phosphorylation and some of the sperm motion parameters, particularly hyperactivation. Inclusion of H89 reduced cAMP stimulation of tyrosine kinase, and tyrosine phosphorylation and motion parameters were reduced almost to basal values. Treatment with genistein reduced tyrosine kinase activity, especially in the soluble fraction of sperm extracts. A decrease in tyrosine phosphorylation of soluble proteins, 105, 81, 55 and 48 kDa, correlated with a significant reduction in sperm motion parameters. Hyperactivation was reduced by tenfold. Tyrosine phosphorylated proteins in the insoluble fraction and the incidence of tyrosine phosphorylated-positive spermatozoa were not reduced markedly. Upstream protein kinase A activation may be a facilitatory rather than an indispensable step in the capacitation-induced tyrosine phosphorylation mediating motility changes in human spermatozoa. Triton-x100 soluble tyrosine phosphorylated proteins, more than their insoluble counterparts, appear to be involved in the modulation of human sperm motion characteristics. PMID- 12887276 TI - Energy metabolism in pig oocytes and early embryos. AB - Pig oocytes and embryos differ from those of other species in having a large quantity of endogenous lipid, a potential role for which has yet to be identified. In the present study, the hypothesis that endogenous triglyceride acts as a metabolic substrate during in vitro maturation and early embryo development was tested. Embryos were produced by in vitro fertilization (IVF) of in vitro-matured, abattoir-derived immature oocytes, cultured in medium NCSU23 up to the blastocyst stage. The triglyceride content of single oocytes and embryos was measured throughout development. Oxygen and glucose consumption and the formation of lactate were measured non-invasively over the same period, enabling total ATP production to be calculated. The triglyceride content of oocytes before maturation (135+/-4.9 ng) decreased by 13 ng (P<0.05) during in vitro maturation, but there was no apparent change in triglyceride content during embryo development (117.68 ng). Oxygen consumption was low throughout embryo cleavage before reaching a peak at the blastocyst stage (P<0.01), a pattern similar to that seen in other mammals studied. Glucose consumption and lactate production were also at a maximum at the blastocyst stage (P<0.05). These data indicate that pig oocytes may use endogenous triglyceride as an energy source during in vitro maturation and that most (91-97%) of the ATP produced during embryo development comes from oxidative phosphorylation. The high exogenous glucose concentration in NCSU23 (5.5 mmol l(-1)) may be needed to form pyruvate, which in turn, produces oxaloacetate, which is required to prime the tricarboxylic acid cycle. However, the reason for the high lipid content in early pig embryos remains to be elucidated. PMID- 12887277 TI - Dose-response of RAG2-/-/gammac-/- mice to busulfan in preparation for spermatogonial transplantation. AB - Practical applications of spermatogonial transplantation require good rates of colonization by the donor cells. Recipient testes are usually depleted of competing endogenous spermatogonia by administration of 32-44 mg busulfan kg(-1) body weight before transplantation. However, it is not clear that this is the optimum dose, especially for immunodeficient mice. In the present study, the response of adult RAG2(-/-)/gamma(c)(-/-) (RAG2) male mice to treatment with 10 50 mg busulfan kg(-1) body weight was determined in terms of mortality rates, testicular masses and histology, and colonization of seminiferous tubules by transplanted spermatogonia. Mortality increased from 0 to 50% at doses between 20 mg busulfan kg(-1) and 40 mg busulfan kg(-1), whereas the maximum effects on testicular mass and histology were observed at 20 mg busulfan kg(-1). Colonization of testes by genetically marked spermatogonia after treatment of mice with 20 mg busulfan kg(-1) was equivalent to rates previously reported in recipients treated with 32-44 mg busulfan kg(-1). Thus, 20 mg busulfan kg(-1) appears to be the optimum dose for preparing RAG2 mice for spermatogonial transplantation. However, because the steepness of the dose-response curves indicates that direct administration of busulfan is not ideal for this purpose, 15 mg busulfan kg(-1) was administered to pregnant females at various times between day 10.5 and day 16.5 of gestation to determine whether this would deplete the number of germ cells in male offspring. Although there were large variations in testicular mass and histology, no mortality was observed and administration of busulfan at day 10.5 or 12.5 after mating delayed initiation of spermatogenesis, indicating that prenatal administration of busulfan combined with neonatal transplantation might be an effective method for further increasing rates of colonization by donor spermatogonia. PMID- 12887278 TI - Acceleration of maturation of FSH and LH responses to photostimulation in prepubertal domestic hens by oestrogen. AB - Egg laying begins in domestic hens, reared on short daylengths, at about day 147 of age and is advanced by photostimulation after but not before about day 42 of age. The development of this response at day 42 may be facilitated by oestrogen. This hypothesis was investigated in prepubertal hens, reared on short daylengths, by comparing the effects of oestrogen treatment on pituitary and plasma FSH and LH responses to photostimulation (16 h light:8 h dark) for 1 week at days 34 and 54 of age. Oestradiol benzoate (0.5 mg kg(-1)) was injected i.m. on alternate days for 1 week before and after photostimulation. At day 34, pituitary LH content increased after photostimulation but plasma LH and FSH concentrations did not increase. At day 54, pituitary FSH content and plasma FSH and LH concentrations increased after photostimulation, whereas pituitary LH content did not increase. At days 34 and 54, oestrogen treatment decreased pituitary FSH and LH contents but did not block the stimulatory effect of photostimulation on pituitary FSH. At day 34 but not at day 54, photostimulation combined with oestrogen treatment increased plasma FSH and LH concentrations. Plasma LH but not plasma FSH concentration increased after GnRH-I injection at days 34 and 54. These observations are consistent with the hypothesis that, in prepubertal female chickens, maturation of the neuroendocrine mechanism mediating photoinduced FSH and LH release may be mediated by oestrogen. This effect of oestrogen on photoinduced LH release may be mediated by increased GnRH-I release or enhanced pituitary responsiveness to GnRH-I. It is proposed that neuroendocrine mechanisms controlling photoinduced FSH release may involve oestrogen-responsive interactions between pituitary paracrine factors, including activins and follistatin. PMID- 12887279 TI - Localization of mRNA for vascular endothelial growth factor (VEGF), angiopoietins and their receptors during the peri-implantation period and early pregnancy in marmosets (Callithrix jacchus). AB - Implantation of a blastocyst into a receptive endometrium is a prerequisite for successful pregnancy. Angiogenesis is a key event in this process but the mechanisms by which localized changes in vascular permeability and angiogenesis occur have yet to be elucidated. Vascular endothelial growth factor (VEGF) and its receptors VEGFR-1 and VEGFR-2 have been implicated as key players in vascular remodelling and placentation. Angiopoietins also appear to have a significant role in regulation of blood vessel growth, maturation and regression. The aim of this study was to describe the molecular regulation of angiogenesis in the first month of pregnancy in marmosets and to address the putative physiological roles for these factors. Uteri were studied at weeks 2, 3 and 4 of pregnancy and compared with late secretory non-pregnant endometrium. Implantation in marmosets occurs at day 11 of pregnancy; hence, these time points were chosen so that the peri-implantation period and very early pregnancy could be studied. mRNAs for VEGF, VEGFR-1 and VEGFR-2, angiopoietin 1, angiopoietin 2 and their receptor Tie 2 were localized and quantified by in situ hybridization. Endothelial cells were identified by CD31 immunocytochemistry. VEGF mRNA was present in all compartments except endothelial cells, and its expression generally increased throughout pregnancy except in upper zone glandular epithelium and luminal epithelium, where a decrease in expression was observed. VEGF receptor mRNAs were found in endothelial cells of the upper zones immediately surrounding glandular epithelium. Angiopoietin 1 mRNA was localized to glandular epithelium of the upper and lower zones throughout pregnancy, and increased in stroma at week 4. Expression of angiopoietin 2 mRNA was localized exclusively to endothelial cells of large luminal vessels and was higher in endometrium from marmosets at week 4 of pregnancy than in endometrium from all other stages. These data provide comprehensive evidence that VEGFR-1 and -2, and angiopoietin 1, angiopoietin 2 and Tie-2 interactions may be involved in the preparation of endometrium for implantation, remodelling of the maternal vasculature and trophoblast invasion during the peri-implantation period in this primate species. PMID- 12887280 TI - Developmental changes in localization of steroid synthesis enzymes in camelid placenta. AB - The uninucleate trophoblast of epitheliochorial camelid placenta produces multinucleate giant cells starting between day 30 and day 35 of pregnancy. The giant cells are found scattered along the trophoblast at similar frequency throughout gestation. Light microscope immunocytochemistry indicates that the four steroid synthesis enzymes (cholesterol side chain cleavage, 3beta hydroxysteroid de-hydrogenase, 17alpha-hydroxylase and aromatase) are present in all uninucleate trophoblast cells between day 14 and day 30 of pregnancy, but are found only in the giant cells once they are established, and that this localization persists until term. The giant cells show massive amounts of smooth endoplasmic reticulum and numerous small mitochondria, again as has been shown in other steroid-producing cells. As progesterone from the corpus luteum is necessary throughout gestation in camels, the capacity for oestrogen production by the trophoblast presumably has an important role, but one limited mostly to the immediate vicinity of the fetomaternal interface. PMID- 12887281 TI - Ovarian development in intrauterine growth-retarded and normally developed piglets originating from the same litter. AB - Epidemiological studies in humans linking adult disease to growth in utero indicate that prenatal life is a critical period for the appropriate development of the reproductive axis. The aim of this study was to compare ovarian development in intrauterine growth-retarded and normally grown piglets originating from the same litter. Intrauterine growth-retarded piglets (runts) were identified on the basis of statistical analysis of the birth weight distribution within each litter. At birth, ovaries were collected from runt piglets (n=14) and their respective mean weight (normal, n=14) littermates. Ovaries were weighed and fixed, and development of ovarian germ cells was quantified in haematoxylin-eosin-stained paraffin wax sections using an image analysis system. Germ cell loss, using an in situ TdT-mediated dUTP nick-end labelling (TUNEL) assay for DNA fragmentation, and follicle cell activity, using immunohistochemistry to demonstrate vimentin, were studied in ovarian sections. At birth, body weight and absolute ovarian mass were significantly lower in runt piglets compared with their respective normally grown littermates (body weight: 733+/-38.5 versus 1530+/-39.7 g; ovarian mass: 51+/-3.0 versus 108+/-9.6 mg; P<0.001 for both). In the ovary, the proportion of nests of oogonia, the number of oocytes and TUNEL-positive cells, and the localization and intensity of vimentin immunoreactivity were not different between runt and normal littermates. However, runt piglets had more primordial follicles (268+/-18.6 versus 235+/-20.1 per mm(2) of cortex; P<0.05), fewer primary follicles (11+/-2.0 versus 20+/-3.0 per mm(2) of cortex; P<0.001) and no secondary follicles compared with normal piglets. These findings indicate that intrauterine growth retardation delayed follicular development in pig ovaries at birth. PMID- 12887282 TI - Motility of spermatozoa at surfaces. AB - The hydrodynamic basis for the accumulation of spermatozoa at surfaces has been investigated. The general conclusion is that when spermatozoa arrive at a surface, they will remain there if the vector of the time-averaged thrust is directed towards that surface. This can arise in two basic ways. First, consider spermatozoa that maintain a three-dimensional waveform and roll (spin) as they progress: in this case, it is argued that the conical (rather than cylindrical) shape of the flagellar envelope will establish the direction-of-thrust necessary for capture by the surface. (Additional findings, for spermatozoa of this type, are that the swim-trajectory is curved and that the direction of its curvature reveals the roll-direction of the cell.) Second, consider spermatozoa that maintain a strictly two-dimensional waveform at the surface: in this case, spermatozoa can be captured because the plane-of-flattening of the sperm head is tilted slightly relative to the plane of the flagellar beat. The sperm head is acting as a hydrofoil and, in one orientation only, it comes to exert a pressure against the surface. (This pressure may possibly, in vivo, aid the penetration of the zona pellucida.) The hydrofoil action of sperm heads may explain any bias in the circling direction of spermatozoa that execute two-dimensional waves at surfaces. Finally, a more complex phenomenon is where interaction of the spermatozoa with the surface appears to induce a three-dimensional to two dimensional conversion of the flagellar wave (thus permitting the hydrofoil effect described). This is characteristic of sperm with 'twisted planar' rather than helical waves. In mammalian spermatozoa, approximately half the beat cycle is planar and the other half generates a pattern of torque causing the head to roll clockwise (seen from ahead), producing a torsion of the neck region of the flagellum. It is the gradual suppression of this torsion, by either impedance at the solid boundary or by raised viscosity, that converts the 'twisted planar' shape into a planar wave. PMID- 12887283 TI - Proopiomelanocortin, a polypeptide precursor with multiple functions: from physiology to pathological conditions. AB - Proopiomelanocortin (POMC) is the polypeptide precursor of ACTH. First discovered in anterior pituitary corticotroph cells, it has more recently been revealed to have many other physiological aspects. The fine molecular mechanisms of ACTH biosynthesis show that ACTH is but one piece of a puzzle which contains many other peptides. Present in various tIssues, among which are pituitary, hypothalamus, central nervous system and skin, POMC undergoes extensive post translational processing. This processing is tIssue-specific and generates, depending on the case, various sets of peptides involved in completely diverse biological functions. POMC expressed in corticotroph cells of the pituitary is necessary for adrenal function. Recent developments have shown that POMC expressing neurons in the brain play a major role in the control of pain and energy homeostasis. Local production of POMC-derived peptides in skin may influence melanogenesis. A still unknown function in the placenta is likely.POMC has become a paradigmatic polypeptide precursor model illustrating the variable roles of a single gene and its various products in different localities. PMID- 12887284 TI - Intrinsic imperfections of endocrine replacement therapy. AB - Hormonal substitution therapy has been extremely successful, with respect to morbidity and mortality, in the treatment of the major syndromes of endocrine insufficiency. However, many patients treated for endocrine insufficiencies still suffer from more or less vague complaints and a decreased quality of life. It is likely that these complaints are, at least in part, caused by intrinsic imperfections of hormone replacement strategies in mimicking normal hormone secretion. Unfortunately, these complaints are often difficult to assess by clinicometric or biochemical tests, because the effects of hormones in general, and thus of hormone replacement strategies in particular, are difficult to quantify at the tIssue level. Therefore, in clinical practice we rely mostly on plasma variables - 'plasma endocrinology' - which are a poor reflection of hormone action at the tIssue level. Appreciation of these intrinsic shortcomings of endocrine therapy is of utmost importance to prevent incorrect labelling of the complaints of many endocrine patients and to achieve further improvement in endocrine replacement strategies. PMID- 12887285 TI - Beta-cell apoptosis in the pathogenesis of human type 2 diabetes mellitus. PMID- 12887286 TI - Evolution of thyroid autoimmunity during iodine prophylaxis--the Sri Lankan experience. AB - OBJECTIVE: To study the evolution of thyroid autoimmunity, in relation to the change in goitre prevalence, during 3 Years of iodine prophylaxis in Sri Lanka. METHODS: Two groups of Sri Lankan schoolgirls between the ages of 10.8 and 17.5 Years were studied in 1998 (401 girls) and 2001 (282 girls). A prospective study was performed in 42 schoolgirls who were thyroid autoantibody (Ab)-positive (+ve) in 1998. Anthropometric measures, urinary iodine excretion (UIE), thyroid Volume, free thyroxine, free tri-iodothyronine, TSH, and thyroglobulin (Tg) and thyroid peroxidase (TPO) Ab were evaluated in all 683 girls. RESULTS: Goitre prevalence was significantly lower in 2001 compared with 1998 related to age (2.9% compared with 20.2%) and body surface area (11.6% compared with 40.8%), although UIE was unchanged. Prevalence of thyroid Ab in 2001 was also lower (23.4% compared with 49.9%); among those with the Ab, 34.8% had TgAb alone and 46.9% had a combination of TgAb+TPOAb, compared with 82.0% TgAb alone in 1998. In 2001, subclinical hypothyroidism was more frequent in Ab+ve (6.3%) than Ab-negative girls (1.0%). A cohort of 42 Ab+ve schoolgirls in 1998 (34 with TgAb alone, eight with TgAb+TPOAb) were evaluated again in 2001. Only 10 of them (23.8%) remained Ab+ve (mostly TPOAb+/-TgAb) in 2001. CONCLUSIONS: This study demonstrates that: (1) in 2001, goitre prevalence and thyroid autoimmunity rates were significantly lower than in 1998; (2) the pattern of thyroid Ab was different in the two surveys; (3) in 2001 alone, the occurrence of hypothyroidism was correlated with the presence of thyroid autoimmunity. These results indicate an evolution of thyroid autoimmune markers during the course of iodine prophylaxis, which has not been described before. PMID- 12887287 TI - Circulating levels of ghrelin in human fetuses. AB - OBJECTIVE: Ghrelin is a GH secretagog isolated recently from rat stomach and involved in the stimulation of food intake and adiposity in rodents and humans. Moreover, subsequent studies showed that ghrelin is expressed in rat and human placenta, suggesting a possible influence of the peptide on fetal growth. The aim of this study was to evaluate circulating levels of ghrelin in appropriate for gestational age (AGA) or intrauterine growth-restricted (IUGR) fetuses. SUBJECTS AND METHODS: Ghrelin levels between 20 and 39 weeks of gestation were measured in 16 AGA and nine IUGR fetuses in whom blood was collected by cordocentesis performed for prenatal diagnosis of different diseases or during elective cesarean section. In most samples, GH, cortisol and leptin levels were also evaluated. Results are expressed as means+/-S.D. Differences were tested using the Student's t-test with Welch correction. P<0.05 was considered significant. RESULTS: All fetuses showed levels of ghrelin in the umbilical venous blood (100+/-99 pmol/l) that did not correlate with the gestational age or the maternal ghrelin levels. No difference was found between umbilical venous and arterial concentrations, suggesting that fetal tIssues are a source of ghrelin. Ghrelin levels in IUGR fetuses were significantly higher than those found in AGA fetuses (176+/-125 vs 58+/-44 pmol/l; P<0.005). Moreover, in samples obtained at birth, ghrelin concentrations correlated negatively with birth weight (P<0.05). In IUGR fetuses, GH and cortisol concentrations were higher and leptin levels lower than in AGA fetuses, although no significant correlation between these parameters and ghrelin levels was found. CONCLUSION: The presence of ghrelin in the fetal circulation as well as its increase in IUGR fetuses suggest a role of this peptide during intrauterine development. PMID- 12887288 TI - Comparison between insulin tolerance test, growth hormone (GH)-releasing hormone (GHRH), GHRH plus acipimox and GHRH plus GH-releasing peptide-6 for the diagnosis of adult GH deficiency in normal subjects, obese and hypopituitary patients. AB - OBJECTIVE: It has been gradually realized that GH may have important physiological functions in adult humans. The biochemical diagnosis of adult GHD is established by provocative testing of GH secretion. The insulin-tolerance test (ITT) is the best validated. The ITT has been challenged because of its low degree of reproducibility and lack of normal range, and is contra-indicated in common clinical situations. Furthermore, in severely obese subjects the response to the ITT frequently overlaps with those found in non-obese adult patients with GHD. DESIGN: The aim of the present study was to evaluate the diagnostic capability of four different stimuli of GH secretion: ITT, GHRH, GHRH plus acipimox (GHRH+Ac), and GHRH plus GHRP-6 (GHRH+GHRP-6), in two pathophysiological situations: hypopituitarism and obesity, and normal subjects. METHODS: Eight adults with hypopituitarism (four female, four male) aged 41-62 Years (48.8+/-1.4 Years), ten obese normal patients (five female, five male) aged 38-62 Years (48.1+/-2.5 Years), with a body mass index of 34.2+/-1.2 kg/m(2), and ten normal subjects (five female, five male) aged 33-62 Years (48.1+/-2.8 Years) were studied. Four tests were performed on each patient or normal subject: An ITT (0.1 U/kg, 0.15 U/kg for obese, i.v., 0 min), GHRH (100 microg, i.v., 0 min), GHRH (100 microg, i.v., 0 min) preceded by acipimox (250 mg, orally, at -270 min and 60 min) (GHRH+Ac); and GHRH (100 microg, i.v., 0 min) plus GHRP-6 (100 microg, i.v., 0 min) (GHRH+GHRP-6). Serum GH was measured by radioimmunoassay. Statistical analyses were performed by Wilcoxon rank sum and by Mann-Whitney tests. RESULTS: After the ITT the mean peak GH secretion was 1.5+/-0.3 microg/l for hypopituitary, 10.1+/-1.7 microg/l (P<0.05 vs hypopituitary) for obese and 17.8+/-2.0 microg/l (P<0.05 vs hypopituitary) for normal. GHRH-induced GH secretion was 2+/-0.7 microg/l for hypopituitary, 3.9+/-1.2 microg/l (P=NS vs hypopituitary) for obese and 22.2+/-3.8 microg/l (P<0.05 vs hypopituitary) for normal. After GHRH+Ac, mean peak GH secretion was 3.3+/-1.4 microg/l for hypopituitary, 14.2+/-2.7 microg/l (P<0.05 vs hypopituitary) for obese and 35.1+/ 5.2 microg/l (P<0.05 vs hypopituitary) for normal. GHRH+GHRP-6 induced mean peak GH secretion of 4.1+/-0.9 microg/l for hypopituitary, 38.5+/-6.5 microg/l (P<0.05 vs hypopituitary) for obese and 68.1+/-5.5 microg/l (P<0.05 vs hypopituitary) for normal subjects. Individually considered, after ITT, GHRH or GHRH+Ac, the maximal response in hypopituitary patients was lower than the minimal response in normal but higher than the minimal response in obese subjects. In contrast, after GHRH+GHRP-6 the maximal response in hypopituitary patients was lower than the minimal response in normal and obese subjects. CONCLUSIONS: This study suggests that, in this group of patients, although both acipimox and GHRP-6 partially reverse the functional hyposomamotropism of obesity after GHRH, but are unable to reverse the organic hyposomatotropism of hypopituitarism, the combined test GHRH+GHRP-6 most accurately distinguishes both situations, without the side effects of ITT. PMID- 12887289 TI - A survey of pituitary incidentaloma in Japan. AB - OBJECTIVE: The development of computed tomography (CT) and magnetic resonance imaging (MRI) has resulted in the discovery of unsuspected endocrinologically silent pituitary masses (pituitary incidentalomas). The aim of this study was to perform a national survey on pituitary incidentalomas in order to establish an appropriate approach to them. DESIGN AND METHODS: Five hundred and six patients with pituitary incidentalomas were obtained by questionnaire from March 1999 to May 2000 under the auspices of the Ministry of Health, Labor and Welfare in Japan. Two hundred and fifty-eight patients underwent surgery (surgical group), while 248 patients were followed up conservatively for a mean period of 26.9 Months (range 6-173 Months) (non-surgical group). Clinical and biochemical assessment, CT or MRI of the pituitary, and visual field testing by Goldman perimetry were assessed at baseline and 6 Months and Yearly thereafter. RESULTS: Thirty-three patients with pituitary incidentalomas (13.3%) developed tumor enlargement during the mean follow-up period of 45.5 Months. Of 115 estimated non functioning adenomas, 23 tumors (20.0%) increased during a mean follow-up period of 50.7 Months (range 10-173 Months), while 5 of 94 (5.3%) estimated Rathke's cysts increased in size during follow-up. Pituitary apoplexy occurred in one of 248 patients (0.4%). CONCLUSIONS: Pituitary incidentalomas usually follow a benign course. We recommend transsphenoidal adenectomy for a solid mass attached to the optic chiasma estimated to be a pituitary adenoma by MRI. Other patients should be followed up by MRI every 6 Months for the first 2 Years, and then Yearly. PMID- 12887290 TI - Expression of tumor necrosis factor (TNF)-alpha protein in the subcutaneous and visceral adipose tissue in correlation with adipocyte cell volume, serum TNF alpha, soluble serum TNF-receptor-2 concentrations and C-peptide level. AB - OBJECTIVE: The aim of the study was to evaluate the expression of tumor necrosis factor (TNF)-alpha protein in the subcutaneous and visceral adipose tIssue in correlation with adipocyte cell Volume, serum TNF-alpha, soluble TNF-receptor-2 (sTNFR-2) and indirect parameters of insulin resistance in overweight/obese and lean healthy persons. DESIGN: A cross-sectional case-control study was used. PATIENTS: Twenty-eight overweight/obese probands with normal glucose tolerance (BMI>27 kg/m(2)) and 15 lean people (BMI<25 kg/m(2)), all of them undergoing planned surgical operation, participated in the study. METHODS: Two to four grams of subcutaneous and visceral adipose tIssue were removed and studied using semi quantitative immunohistochemical staining of the TNF-alpha protein. Serum TNF alpha, sTNFR-2 (ELISA) and fasting C-peptide (RIA) were measured. RESULTS: TNF alpha protein was expressed in adipocytes of both depots. The expression was evaluated visually and found to be greater in the obese patients. Significantly higher serum TNF-alpha (5.58+/-0.87 pg/ml vs 4.21+/-0.55, mean+/-s.d., P<0.01, Mann-Whitney) and sTNFR-2 levels (7.84+/-3.56 ng/ml vs 4.59+/-1.35, P=0.005) were found in the obese subgroup in correlation with the fasting C-peptide level (r=0.49, P=0.003; and r=0.74, P=0.001) and the C-peptide/ blood glucose ratio (r=0.47, Spearman, P=0.005; and r=0.70, P=0.001). The cell Volume of both adipocyte depots was found to have a significant positive correlation with serum TNF-alpha and sTNFR-2 levels in the total group of patients (subcutaneous: r=0.52, P=0.0003; r=0.69, P<0.0001; visceral: r=0.65, P<0.0001; r=0.63, P<0.0001) and in both subgroups. CONCLUSIONS: Adipocyte cell Volume of both the subcutaneous and visceral fat depots may be determinants of TNF-alpha, sTNFR-2 production and obesity-linked insulin resistance. PMID- 12887291 TI - Mutational spectrum of congenital adrenal hyperplasia in Slovenian patients: a novel Ala15Thr mutation and Pro30Leu within a larger gene conversion associated with a severe form of the disease. AB - OBJECTIVE: To analyse the mutational spectrum, the associated haplotypes and the genotype-phenotype correlation, and to design a reliable and rational approach for CYP21 mutation detection in Slovenian congenital adrenal hyperplasia (CAH) patients. DESIGN: Molecular analysis of the CYP21 gene was performed in 36 CAH patients and 79 family members. METHODS: Southern blotting, sequence-specific PCR amplification (PCR-SSP), sequence-specific oligonucleotide hybridisation (PCR SSO) and sequencing were used to detect CYP21 gene deletions, conversions and point mutations. RESULTS: CYP21 gene deletion was the most frequent mutation (36.4%). Large gene conversions detectable only by Southern blotting represented 12.1%, and gene conversions involving the promoter region represented 7.6% of the mutated alleles. The most frequent point mutations were: intron 2 splice mutation 16.7%, Ile172Asn mutation 7.6%, Gln318Stop 7.5% and Pro30Leu 12.2% of alleles. A correlation between the genotype and the clinical phenotype similar to those described for large populations was observed. The finding of Pro30Leu mutation linked to a gene conversion could explain the simple virilising (SV) phenotype in compound heterozygotes for the Pro30Leu and a severe mutation. In two siblings with a salt wasting form of CAH (SW-CAH), a novel mutation Ala15Thr was found on the allele characterised by Pro30Leu mutation and gene conversion involving the promoter region. CONCLUSIONS: Our genotyping approach allowed reliable diagnosis of CAH in the Slovenian population. The high frequency of CYP21 gene aberrations on Pro30Leu positive alleles justified systematic searching for a gene conversion in the promoter region using the PCR-SSP reaction. PMID- 12887292 TI - The associations of age, lifestyle factors and chronic disease with testosterone in men: the Tromso Study. AB - OBJECTIVE: To study whether lifestyle factors and/or chronic disease are associated with the age-related decline of total and free testosterone in men, or if these factors might be associated with the variation of total and free testosterone but not with their age-related decline. DESIGN: A population-based, cross-sectional study was used. METHODS: Total testosterone and sex hormone binding globulin (SHBG) levels were analyzed and free testosterone levels were calculated in 1563 men participating in the Tromso study in 1994/1995. Anthropometric characteristics were also measured and two standardized questionnaires completed, including lifestyle factors and medical history. The data were analyzed with multiple linear regression analysis of covariance, and logistic regression. RESULTS: Total and free testosterone were inversely associated (P=0.001 and P<0.001), while SHBG was positively associated (P<0.001) with age. Body mass index (BMI) was inversely associated with total (P<0.001) and free (P=0.016) testosterone and SHBG (P<0.001). Both total and free testosterone were positively associated with tobacco consumption (P<0.001 and P=0.004) and total testosterone was positively associated with coffee consumption (P<0.001). SHBG was positively associated with smoking (P=0.004) and coffee consumption (P<0.001). Men who reported having had a stroke or having a cancer diagnosis had lower levels of total testosterone (P<0.001 and P<0.01) and free testosterone (P<0.01). CONCLUSIONS: BMI and smoking are independent contributors to the variation of total and free testosterone and SHBG levels, and coffee consumption to the variation of total testosterone and SHBG. Thus, lifestyle factors can have a direct effect on circulating levels of free endogenous sex hormones and to total levels due to the effect on SHBG levels. PMID- 12887293 TI - Enhanced anterior pituitary mitotic response to adrenalectomy after multiple glucocorticoid exposures. AB - OBJECTIVES: Glucocorticoid withdrawal in man is associated with transient but sometimes prolonged impairment of hypothalamo-pituitary-adrenal axis secretory responsiveness. This has led to continued concern in the clinical arena. The acute anterior pituitary response to glucocorticoids in the rat includes apoptosis-mediated deletion of a cell population. Whilst continued cell turnover following glucocorticoid withdrawal and the potential for differentiation of uncommitted precursor cells and transdifferentiation of other secretory cell types predicts that, given sufficient time, complete anterior pituitary trophic recovery is likely, this hypothesis has not previously been tested. DESIGN AND METHODS: We have quantified pituitary mitotic and apoptotic rate, as well as corticotroph number and pro-opiomelanocortin transcript prevalence, together with hypothalamic corticotrophin-releasing hormone and vasopressin transcript prevalence 5 weeks after three short dexamethasone treatments each separated by a week. Bilateral adrenalectomy was then carried out as a maximal secretory and trophic stimulus, and the response to a fourth dexamethasone treatment assessed 1 week later. RESULTS: Anterior pituitary mitotic index was significantly higher in rats previously exposed to dexamethasone compared with age-matched controls exposed to dexamethasone for the first time. No differences were found in the subsequent apoptotic response to a fourth dexamethasone treatment or in the levels of paraventricular corticotrophin-releasing hormone or vasopressin transcripts or pituitary pro-opiomelanocortin transcripts. CONCLUSIONS: These data indicate that full recovery of pituitary mitotic activity takes longer than the recovery of secretory parameters, and suggest that, for several weeks after glucocorticoid exposure, the ability of the pituitary to meet fresh demands placed on it may be suboptimal. PMID- 12887294 TI - Wellcome Trust Award Lecture. Chromatin-modifying enzymes in transcription and cancer. AB - In recent years, our view of how gene expression is controlled has changed dramatically. The discovery of enzymes that modify histones has revealed that transcription is an enzymically driven process. Such modifications can recruit specific proteins and mediate chromatin changes that affect transcription either positively or negatively. Important biological pathways leading to cell proliferation are under the control of these enzymes, and several of them are found deregulated in cancer. The hope is that chromatin-modifying enzymes will be a rich source of targets for drug discovery. PMID- 12887295 TI - Introduction to mannan-binding lectin. AB - Mannan-binding lectin (MBL) was first discovered as a plasma opsonin for baker's yeast and was independently characterized biochemically. It belongs to the small subfamily of collectins: C-type lectins possessing a collagen-like domain. MBL is synthesized by the liver and secreted into the bloodstream. It is believed to be an important component of innate immunity, acting as an ante-antibody and/or as a disease modifier. It is thought to influence disorders as diverse as meningococcal disease, rheumatoid arthritis, cystic fibrosis and recurrent miscarriage. Lack of MBL may be most relevant in the context of a co-existing secondary immune deficiency. Replacement therapy, first carried out 30 years ago with unfractionated plasma, appears promising. The development of a recombinant product should permit the extension of MBL therapy to randomized clinical trials of sufficient size to provide clear evidence about the physiological significance of this intriguing glycoprotein. PMID- 12887296 TI - Biochemistry and genetics of mannan-binding lectin (MBL). AB - Mannose- or mannan-binding lectin (MBL) is a member of the collectin protein family, which includes lung surfactant proteins SP-A and SP-D. Each member consists of similar or identical polypeptide chains with a region of collagen like sequence followed by a C-type lectin domain. The polypeptides associate in threes to form a subunit containing a collagen-like helix, with three clustered lectin domains. These subunits associate into larger structures, usually with 12 18 polypeptides. The collectins bind to patterns of neutral sugars on surfaces (e.g. of micro-organisms) and mediate effector functions associated with killing/phagocytosis. MBL is the only collectin which activates complement. It resembles in quaternary structure the complement protein C1q, which recognizes targets via charge clusters. Binding of MBL to a surface activates MBL-associated serine proteases (MASPs) attached to MBL, and MASP-2 activates complement proteins C4 and C2. The MASPs are homologous to the C1q-associated proteases, C1r and C1s. MBL therefore activates complement by a mechanism very similar to C1q, and engages the opsonic activity of complement to clear micro-organisms. The serum concentration of MBL is very variable in humans. The variability is largely associated with mutations leading to amino acid substitutions in the collagen like region which decrease MBL assembly and stability. Many studies demonstrate that MBL deficiency is associated with susceptibility to a range of infectious and inflammatory diseases. PMID- 12887297 TI - Anti-microbial activities of mannose-binding lectin. AB - Mannose-binding lectin (MBL; also known as mannan-binding lectin) is involved in first-line defence by binding to bacteria, viruses, protozoa and helminths through a pattern-recognition mode of detection and then initiating a range of host responses. Currently, we have been unable to extrapolate from what we know of the biochemistry of MBL binding to predict accurately the interaction of MBL with individual micro-organisms; even subtle surface alterations have been shown to have an extensive impact on MBL-mediated recognition of pathogens. MBL has a major protective effect through activation of the complement system via MBL associated serine proteases (MASPs). This can cause the lysis of Gram-negative bacteria and also opsonize a wide spectrum of potential pathogens for phagocytosis. MBL may also influence phagocytosis in the absence of complement activation through an interaction with one or more collectin receptors. This may also be the basis for a direct effect of the protein on inflammatory responses. MBL can alter the function of microbial structures, such as gp120 of HIV, to prevent infection. The protein may also interact with the components of other cascade systems such as the clotting system, which will have a role in microbial pathogenesis. An understanding of these basic mechanisms will be vital if we are to use purified or recombinant MBL in therapeutic applications. PMID- 12887298 TI - Mannan-binding lectin (MBL) production from human plasma. AB - Individuals with low levels of mannan-binding lectin (MBL) appear to be susceptible to infectious diseases. This suggests that substitution therapy with MBL might be a beneficial treatment of patients with MBL deficiency. A production process for an MBL product has been developed from a fraction II+III precipitate obtained by ethanol fractionation of plasma. The MBL process includes three chromatographic steps, where the first and key step is affinity chromatography on a cross-linked agarose matrix selecting for oligomeric, carbohydrate-binding MBL. The yield from the production process is about 25% of the plasma MBL content, and the purity is about 65%. The MBL product shows mannan-binding activity and complement-activating ability. A safety study has shown this plasma-derived MBL to be safe and well tolerated in adult MBL-deficient volunteers. PMID- 12887299 TI - Recombinant mannan-binding lectin (MBL) for therapy. AB - Mannan-binding lectin (MBL) is a plasma protein involved in the innate immune response. It binds to a number of micro-organisms and promotes killing of these through complement activation either directly or through opsonization. Clinical evidence indicates that in a variety of situations genetically determined low MBL levels are associated with increased susceptibility to infections. Infusions of plasma-derived MBL into MBL-deficient individuals was found to be safe in preliminary trials, but we considered that sufficient production and product safety could only be achieved through synthesis of recombinant MBL. A transfected human cell line produces MBL showing the same biological activity as plasma derived MBL, and an essentially identical profile on MS. The production has been scaled up and clinical trials will start this year. PMID- 12887300 TI - Infusion of plasma-derived mannan-binding lectin (MBL) into MBL-deficient humans. AB - Our first experience of mannan-binding lectin (MBL)-replacement therapy was with a patient experiencing recurrent erythema multiforme associated with reactivation of herpes simplex virus; his erythematous eruptions could be controlled with infusions of fresh frozen plasma containing MBL, but not with plasma lacking MBL. Some years later, we treated a young girl with recurrent, debilitating infections with purified MBL; this was also followed by a dramatic clinical improvement. We have now carried out a phase I clinical trial on 20 MBL-deficient, but healthy, adult volunteers. The MBL was prepared by the State Serum Institute in Copenhagen, Denmark, from blood donor plasma. Each volunteer received a total of 18 mg of MBL in three 6-mg doses given intravenously once a week over 3 weeks. The volunteers were monitored closely after each infusion and no adverse clinical or laboratory effects were observed. Laboratory parameters included C-reactive protein, various complement components, and antibodies to MBL, HIV and hepatitis viruses. C3a (the anaphylotoxin derived from complement component C3) was monitored for signs of complement activation, but no significant infusion associated fluctuations were observed. Serum levels of MBL after each 6-mg infusion ranged between 1200 and 2500 ng/ml. The half-life of the infused MBL was about 70 h, or 3 days. It was concluded that infusion of purified MBL manufactured by the Danish State Serum Institute is a safe procedure. However, adults may have to be given 6 mg or more at least twice weekly to maintain protective plasma MBL levels in MBL-deficient individuals. PMID- 12887301 TI - Clinical potential of mannose-binding lectin-replacement therapy. AB - Mannose-binding lectin (MBL; also known as mannan-binding lectin) is an important component of innate immunity. MBL levels are mainly genetically determined. Low serum MBL levels and their cognate haplotypes have been associated with a wide range of infections. However, most subjects with MBL deficiency remain healthy. MBL deficiency is also associated with non-infectious diseases including systemic lupus erythematosus, rheumatoid arthritis, cystic fibrosis and common variable immunodeficiency. MBL deficiency may affect susceptibility to (e.g. meningococcal disease), or alter the natural history of (e.g. rheumatoid arthritis, cystic fibrosis), a disease. MBL (plasma-derived or recombinant) therapy has yet to be shown to be safe and effective. Potentially it may be useful in MBL-deficient patients to reduce susceptibility to, or enhance recovery from, bacterial infection or to alter the natural history of a disease (disease-modifying drug). In practise the place of MBL therapy may be as a disease-modifying drug to reduce the severity of rheumatoid arthritis and to preserve lung and liver function in cystic fibrosis. MBL therapy may also ameliorate various immunodeficiency syndromes. A potential hazard of MBL therapy is enhanced complement-mediated host damage. The place of MBL therapy will await results of randomized controlled clinical trials. PMID- 12887302 TI - Mannan-binding lectin (MBL) serum levels and post-operative infections. AB - Mannan-binding lectin (MBL) is a central component of the innate immune system. Here we investigated the role of MBL in surgical patients during the peri operative phase. Basal and post-operative (days 1-3 post-surgery) serum samples were obtained prospectively from 156 patients undergoing major elective gastrointestinal surgery for malignant disease. In contrast to procalcitonin (a typical acute-phase protein), there was no significant difference in serum MBL between pre- and post-operative samples (P=0.62). Nevertheless, patients who developed post-operative infections showed significantly lower pre- and post operative MBL levels than those who did not (P=0.013 and P=0.005, respectively). There was no significant difference in pre-operative procalcitonin between the two groups (P=0.56). We conclude (i) that serum MBL levels did not respond immediately to surgical trauma, and (ii) that lower MBL levels were associated with an increased occurrence of post-operative infections. Studies on larger patient groups are necessary, however, to assess the value of MBL measurements in identifying patients at risk of post-operative complications. PMID- 12887303 TI - Consensus statement on the future of mannan-binding lectin (MBL)-replacement therapy. PMID- 12887304 TI - Psp and Smgb: a model for developmental and functional regulation in the rat major salivary glands. AB - This paper summarizes past work detailing the developmental expression, cell and organ localization and biochemical features of the proteins parotid secretory protein (PSP) and isoforms of submandibular gland protein B (SMGB), and describes the molecular characterization of the genes that encode them, Psp and Smgb. These genes appear to be related to the BPI (bactericidal/permeability-increasing protein)/LBP (lipopolysaccharide-binding protein)/PLUNC (palate, lung and nasal epithelial clone) gene family found in the oral and respiratory organs of humans, rodents and cattle. We have emphasized the diverse patterns of expression of these genes among the submandibular, sublingual and parotid salivary glands of the rat, and their potential usefulness in defining and identifying genomic regulatory mechanisms of salivary development. While Psp is expressed similarly in the mouse, the putative Smgb gene of the mouse seems not to be expressed, apparently due to the insertion, between exons 1 and 2, of a gene for a retroviral protein. PMID- 12887305 TI - Bovine parotid secretory protein: structure, expression and relatedness to other BPI (bactericidal/permeability-increasing protein)-like proteins. AB - Members of the family of BPI (bactericidal/permeability-increasing protein)-like proteins are as yet incompletely characterized, particularly in cattle, where full-length sequence information is available for only three of the 13 family members known from other species. Structural bioinformatic analyses incorporating bovine homologues of several members of the BPI-like protein family, including two forms of bovine parotid secretory protein (PSP), showed that this family is also present in cattle. Expression analyses of several members of the BPI-like protein family in cattle, including PSP (Bsp30), von Ebner's minor salivary gland protein (VEMSGP) and lung-specific X protein (LUNX), showed a restricted pattern of expression, consistent with earlier hypotheses that these proteins function in the innate immune response to bacteria. The possible role of bovine PSP in susceptibility to pasture bloat in cattle is discussed. PMID- 12887306 TI - Bactericidal/permeability-increasing protein (BPI) and lipopolysaccharide-binding protein (LBP): structure, function and regulation in host defence against Gram negative bacteria. AB - Lipopolysaccharide-binding protein (LBP) and bactericidal/permeability-increasing protein (BPI) are closely related endotoxin-binding proteins that function in a co-ordinated manner to facilitate an integrated host response to invading Gram negative bacteria. Differences in the structure and function of BPI and LBP, as well as differences in their mobilization, permit highly sensitive pro inflammatory responses to small numbers of bacteria at the onset of bacterial invasion and, later, efficient elimination of viable bacteria and their remnants and of endotoxin-driven inflammation. PMID- 12887307 TI - Structure of human BPI (bactericidal/permeability-increasing protein) and implications for related proteins. AB - Human bactericidal/permeability-increasing protein (BPI) belongs to a family of mammalian lipopolysaccharide-binding and lipid transport proteins. Recent sequence database searches indicate that several other protein families, including the palate, lung and nasal epithelial clone (PLUNC), parotid secretory protein (PSP) and BPI-like proteins, are likely to share the BPI fold, which was determined through X-ray crystallographic studies. As the single representative of its fold family of known structure, the three-dimensional model of BPI suggests structural features that are likely to be conserved across this large and varied group of proteins. PMID- 12887308 TI - Expression of BPI (bactericidal/permeability-increasing protein) in human mucosal epithelia. AB - Among the antimicrobial proteins and peptides of humans is the cationic 55 kDa bactericidal/permeability-increasing protein (BPI), which possesses antibacterial, endotoxin-neutralizing and opsonic activity against Gram-negative bacteria. Although identified originally as an abundant constituent of neutrophil granules, we have recently identified functional expression of BPI by human mucosal epithelia. BPI expression was markedly up-regulated by exposure of epithelia to lipoxins, endogenous anti-inflammatory eicosanoids that are generated in vivo in the context of aspirin treatment (aspirin-triggered lipoxins). Epithelial BPI was found to be surface expressed and fully functional, as measured by antibacterial activity against Salmonella typhimurium as well as lipopolysaccharide (LPS; endotoxin)-neutralizing activity. These results suggest a role for BPI as an effector of epithelial antibacterial activity and as a modulator of epithelial responses to LPS. Both BPI and the lipoxins are currently the subject of intensive biopharmaceutical development, raising the possibility that therapeutic use of BPI or modulation of epithelial BPI expression may be a useful adjunctive therapy for conditions in which epithelial inflammation is associated with Gram-negative infections and/or endotoxin. PMID- 12887309 TI - Four BPI (bactericidal/permeability-increasing protein)-like genes expressed in the mouse nasal, oral, airway and digestive epithelia. AB - A cluster of related genes whose products show structural identity with bactericidal permeability-increasing protein (BPI) has been identified in the genomes of both mice (on chromosome 2) and humans (on chromosome 20). Genes in the cluster include those encoding parotid secretory protein (PSP), von Ebner minor salivary gland protein (VEMSGP) and sequences in the PLUNC (palate, lung and nasal epithelium clone) family, among others. This mini-review addresses the tissue-specific expression of these genes in the mouse. PMID- 12887310 TI - Comparative analysis of the PLUNC (palate, lung and nasal epithelium clone) protein families. AB - PLUNC (palate, lung and nasal epithelium clone) is a small, secreted protein that is expressed in the oropharynx and upper airways of humans, mice and rats. We have described a family of at least 14 PLUNC genes localized on chromosome 20 (in humans), 2 (in mice) or 3 (in rats). These rapidly evolving proteins are structurally related to lipopolysaccharide-binding protein (LBP) and bactericidal/permeability-increasing protein (BPI). In the present analysis we comment on the comparative aspects of this protein family, which may function to protect epithelial surfaces from pathogenic micro-organisms. PMID- 12887311 TI - PLUNC (palate, lung and nasal epithelial clone) proteins in human nasal lavage fluid. AB - PLUNC (palate, lung and nasal epithelial clone) is a newly discovered gene that is expressed in the upper respiratory tract and is suggested to be of importance in host defence against bacteria. We have identified two forms of the PLUNC protein in human nasal lavage fluid (NLF) using two-dimensional gel electrophoresis (2-DE) and MS. The apparent molecular masses and isoelectric points of these forms are 24.8 kDa/pI 5.4 and 25.1 kDa/pI 5.5. Notably, the 24.8 kDa/pI 5.4 form of PLUNC is an abundant protein in the 2-DE protein patterns of NLF from healthy subjects. Decreased levels of PLUNC were found in NLF from smokers and workers exposed to reactive epoxy chemicals, indicating that long term exposure to airway irritants impairs the production of PLUNC in the upper respiratory tract. We have also investigated the presence of lipopolysaccharide (LPS)-binding proteins in NLF. Five proteins were found to adsorb to a LPS-coated surface; two of these proteins correspond to the two PLUNC forms, as judged by 2 DE pattern matching. For comparison, human saliva was found to contain a set of LPS-binding proteins with similar 2-DE spot positions (the same pIs but somewhat lower apparent molecular masses of approximately 20 kDa). These results indicate that PLUNC may be a new marker of airway inflammation and may play a part in the innate immune response, and that human saliva contains yet other members of the family of LPS-binding proteins. PMID- 12887312 TI - Expression and anti-bacterial activity of human parotid secretory protein (PSP). AB - Parotid secretory protein (PSP) is an abundant protein in mouse and rat parotid glands. A related sequence (C20orf70) was identified on human chromosome 20. The goal of this study was to determine if PSP is expressed in the human parotid gland. The cDNA for human PSP was amplified from a human parotid cDNA sample. A peptide antibody, raised to the C-terminal peptide of PSP, identified the protein in human parotid tissue by immunofluorescence microscopy. Immunoaffinity chromatography suggested that PSP was expressed in human saliva. PSP is related to bactericidal/permeability-increasing protein (BPI). To test if PSP exhibits anti-bacterial activity, epitope-tagged PSP was expressed in rat GH4C1 cells. The secretion medium exhibited bacteristatic or bactericidal effects on Pseudomonas aeruginosa in a colony-forming assay when compared with secretion medium from GH4C1 cells that did not express PSP. These results suggest that PSP is expressed in the human parotid gland and saliva, where it functions as a BPI-like anti bacterial protein. PMID- 12887313 TI - Visualizing membrane trafficking using total internal reflection fluorescence microscopy. AB - There is a dizzying array of fluorescent probes now commercially available to monitor cellular processes, and advances in molecular biology have highlighted the ease with which proteins can now be labelled with fluorophores without loss of functionality. This has led to an explosion in the popularity of fluorescence microscopy techniques. One such specialized technique, total internal reflection fluorescence microscopy (TIR-FM), is ideally suited to gaining insight into events occurring at, or close to, the plasma membrane of live cells with excellent optical resolution. In the last few years, the application of TIR-FM to membrane trafficking events in both non-excitable and excitable cells has been an area of notable expansion and fruition. This review gives a brief overview of that literature, with emphasis on the study of the regulation of exocytosis and endocytosis in excitable cells using TIR-FM. Finally, recent applications of TIR FM to the study of cellular processes at the molecular level are discussed briefly, providing promise that the future of TIR-FM in cell biology will only get brighter. PMID- 12887314 TI - Regulation of the exocytotic machinery by cAMP-dependent protein kinase: implications for presynaptic plasticity. AB - For over a decade, the enhancement of regulated exocytosis by cAMP-dependent protein kinase (PKA) has remained unexplained at the molecular level. The fact that this phenomenon has been observed in such a wide variety of secretory cell types, from pancreatic beta-cells to neurons, suggests that it is an important and fundamental mechanism. Extensive analysis of the phosphorylation of exocytotic proteins has yielded few substrates of PKA in vitro, and fewer still have had physiological effects attributed to their phosphorylation. Here we review two proteins that do fulfil these criteria: the synaptic vesicle proteins cysteine string protein (CSP) and Snapin. Phosphorylation of these proteins by PKA produces changes in their respective protein-protein interactions, and has been attributed to modulation of the vesicle priming (Snapin) and vesicle fusion (CSP) stages of exocytosis. We also discuss how the function of CSP and Snapin phosphorylation might fit into an interesting aspect of the PKA-dependent enhancement of exocytosis: presynaptic plasticity in the brain. PMID- 12887315 TI - Neuronal calcium sensor-1: a multifunctional regulator of secretion. AB - Ca2+ ions play a crucial role not only as the trigger for neurotransmitter release, but also in other aspects of brain function, such as short-term and long term modulation of synaptic efficacy, which may underlie certain forms of learning and memory. The actions of Ca2+ are mediated by Ca(2+)-binding proteins, including a group of proteins known as neuronal calcium sensor (NCS) proteins. The NCS family includes NCS-1, visinin-like proteins, recoverins, guanylate cyclase-activating proteins and potassium channel-interacting proteins. Some members of this family, such as recoverin and guanylate cyclase-activating protein, are only expressed in photoreceptor cells and have been implicated in the control of visual transduction pathways, while the functional roles of the other members are largely unknown. NCS-1 was originally identified in Drosophila in a screen for neuronal hyperexcitability mutants. NCS-1 is an N-terminally myristoylated protein that contains four EF-hand motifs, three of which are able to bind Ca2+ in the submicromolar range. Overexpression of NCS-1 has been shown to enhance evoked neurotransmitter release, paired-pulse facilitation and exocytosis in several neuronal and neuroendocrine cell types. Recent experiments suggest that NCS-1 interacts directly with phosphatidylinositol 4-hydroxykinase in yeast as well as mammalian cells, suggesting that it may enhance neuronal secretion by modulating cellular trafficking steps in a phosphoinositide dependent manner. In contrast, an involvement of NCS-1 in the expression and regulation of voltage-gated Ca2+ channels and K+ channels has also been proposed, which may be attributed, at least in part, to the effects of NCS-1 on vesicular trafficking pathways. The present review describes current knowledge about the cellular functions and molecular mechanisms by which NCS-1 may regulate neurotransmitter release. PMID- 12887316 TI - Insulin secretion by 'kiss-and-run' exocytosis in clonal pancreatic islet beta cells. AB - Exocytotic release of neuropeptides and hormones is generally believed to involve the complete merger of the secretory vesicle with the plasma membrane. However, recent data have suggested that 'kiss-and-run' mechanisms may also play a role. To analyse secretory events in neuroendocrine beta-cells, we imaged chimaeric reporters targeted to either the vesicle membrane [chimaeras of synaptobrevin-2 and pH-sensitive green fluorescent protein (synapto.pHluorin) or of phogrin (phosphatase on the granule of insulinoma) and enhanced green fluorescent protein (EGFP) (phogrin.EGFP)] or the lumen [neuropeptide Y (NPY).pH-insensitive yellow fluorescent protein (Venus)] by evanescent wave microscopy. Unexpectedly, the frequency of NPY.Venus release events was only 17-27% of that of vesicle fusion reported with synapto.pHluorin, but not phogrin.EGFP, indicating that exocytosis of cargo peptides that is likely to require complete collapse of the vesicle into the plasma membrane is relatively rare. However, both the frequency and the kinetics of NPY.Venus release were modulated by stimulus strength or by overexpression of synaptotagmin IV, demonstrating the plasticity of 'kiss-and run' fusion. PMID- 12887317 TI - Relationship between fusion pore opening and release during mast cell exocytosis studied with patch amperometry. AB - We have studied exocytosis in rat peritoneal mast cells by cell-attached patch amperometry. Step increases in capacitance were accompanied by typical amperometric spikes due to the release of 5-hydroxytryptamine (serotonin), indicating exocytosis of typical mast cell granules. We have measured the time course of fusion pore expansion, and correlated it with release from the granule matrix. The fusion pore of mast cell granules grows in three stages. The initial expansion of the pore occurred at a rate of 5 nS/s, and in many cases an observable amperometric foot was detected. A second, rapid expansion phase occurred with a rate as high as 1000 nS/s, coinciding with the upstroke of the amperometric spike. A third, slower phase, with a rate of 5 nS/s, completed the final expansion of the fusion pore. These data reveal the very late stages in the exocytotic process, and demonstrate that the size of the fusion pore does not limit release during the upstroke of the amperometric spike or during the final, slow expansion that occurs during for the decay of the amperometric spike. PMID- 12887318 TI - Requirement for N-ethylmaleimide-sensitive factor for exocytosis of insulin containing secretory granules in pancreatic beta-cells. AB - N-ethylmaleimide-sensitive factor (NSF) has an important role in fusion processes within intracellular compartments and at the plasma membrane, but the exact role of this protein in the exocytotic machinery has not yet been determined. NSF was found to be present in the cytosol of rat pancreatic beta-cells and rat insulinoma INS-1 cells. Capacitance measurements revealed that exocytosis of primed granules was not affected by the presence of a monoclonal antibody against NSF, mAb 2E5, suggesting that NSF is not involved in the fusion process. The antibody markedly decreased rapid refilling of new granules from a reserve pool during a first stimulation. However, slow refilling of primed granules occurred within a 2 min period between the first and second stimulations. We conclude that NSF is required in the exocytotic process in order to obtain a complete exocytotic response. Possible mechanisms by which NSF takes part in this process in insulin-secreting rat beta-cells are discussed. PMID- 12887319 TI - Role of Munc18-1 in synaptic vesicle and large dense-core vesicle secretion. AB - SNARE (soluble N-ethylmaleimide-sensitive factor attachment protein receptor) complex formation between a vesicle and the target membrane is a central aspect of probably all vesicle fusion reactions. The sec1/munc18 (SM) protein family is also involved in membrane trafficking and fusion events. However, in contrast with the consensus on SNARE protein function, analysis of SM proteins in different systems has produced different ideas about their exact role, their site of action and their relationship to SNARE proteins. Deletion of the SM protein involved in secretory vesicle release in mice, Munc18-1, results in a complete block of exocytosis. Manipulation of Munc18-1 protein levels in neurons and adrenal chromaffin cells argues for a positive role of this protein in vesicle secretion, as overexpression results in an increase in vesicle secretion. A decrease in Munc18-1 protein levels, on the other hand, leads to a decrease in vesicle secretion. PMID- 12887320 TI - Red, yellow, green go!--A novel tool for microscopic segregation of secretory vesicle pools according to their age. AB - Large dense-core vesicles (LDCVs) were labelled in cultured bovine adrenal chromaffin cells expressing fluorescent chimaeric 'cargo' proteins that were targeted to these secretory vesicles. When the cells were stimulated with nicotine 48 h after transduction, the fractional loss of fluorescent LDCVs was much greater than the fractional catecholamine secretion, implying selective release of newly assembled vesicles. This was confirmed using a fluorescent 'timer' construct that changes its fluorescence emission from green to red over several hours, and by measurement of the location and mobility of LDCVs in live cells by confocal fluorescence microscopy. Newly assembled (green) LDCVs were located mostly in peripheral regions of the cells, were virtually immobile and could be released by nicotine, but not by Ba2+; in contrast, older (red) LDCVs were centrally located and relatively mobile, and their exocytotic release was triggered by Ba2+, but not by nicotine. We describe the image restoration procedure that is necessary in order to analyse the behaviour of LDCVs labelled with this construct. PMID- 12887321 TI - Kinases in clathrin-mediated endocytosis. AB - The process of clathrin-coated vesicle (CCV) formation/disassembly has been studied intensively, and numerous proteins have been identified which aid this process. We have learnt a great deal about individual components of the CCV machinery, and now the ultimate aim is to elucidate the mechanisms regulating clathrin-mediated trafficking. One of the fundamental processes governing the complicated network of interactions is phosphorylation. It has been known for some time that several proteins associated with clathrin-coated vesicles are substrates for protein kinases. These proteins include clathrin, three of the four adaptor complex subunits, dynamin 1, synaptojanin 1 and the amphiphysins. However, the identities of the kinases involved in this process remained largely unknown until recently. This short review discusses advances in our knowledge of how CCV formation/disassembly is regulated by the phosphorylation/dephosphorylation cycle and the role played by specific protein kinases in that process. PMID- 12887322 TI - Imaging SNAREs at work in 'unroofed' cells--approaches that may be of general interest for functional studies on membrane proteins. AB - When cultured cells are subjected to a brief ultrasound pulse, their upper parts burst, but the basal plasma membranes with their embedded membrane-protein complexes remain intact. Such two-dimensional, paraformaldehyde-fixed plasma membrane sheets have been used in the past to visualize the morphology of the inner plasmalemmal leaflet by electron or light microscopy. More recently, fluorescence microscopy of unfixed native membranes has been applied to study SNARE (soluble N-ethylmaleimide-sensitive factor attachment protein receptor) function. For instance, biochemical reactions of the plasmalemmal SNAREs with soluble fluorescent SNAREs, patching of SNARE and raft domains, and online monitoring of SNARE-mediated membrane fusion has been performed. The results obtained with the membrane sheet system have added some novel aspects to our understanding of the regulation of neuronal exocytosis. Surprisingly, SNAREs are concentrated in cholesterol-dependent microdomains that are different from membrane rafts. SNAREs in such domains are highly reactive, and define sites for vesicle exocytosis. Secretory granules that fuse on the membrane sheets are retrieved intact in a dynamin-dependent process, suggesting that the 'kiss-and run' mechanism is not a reversed SNARE reaction, but is driven by a biochemically different mechanism. So far, studies of this type have focused on neuronal exocytosis; however, the method might be widely applicable. Data obtained with this system are derived from a 100% pure plasma membrane preparation that is only several seconds old, and membrane proteins are studied in their natural microenvironment that is defined by local lipid composition and putative bound proteins. Hence this approach yields results that most probably reflect the situation in a live cell. PMID- 12887323 TI - Assembly of N-methyl-D-aspartate (NMDA) receptors. AB - The N-methyl-D-aspartate receptor (NMDAR) requires both NR1 and NR2 subunits to form a functional ion channel. Despite the recent advances in our understanding of the contributions of these different subunits to both the function and pharmacology of the NMDAR, the precise subunit stoichiometry of the receptor and the regions of the subunits governing subunit interactions remain unclear. Since NR2 subunits are not transported to the cell surface unless they associate with NR1 subunits, cell-surface expression of NR2A can be used to monitor the association of the different subunits in cells transfected with N- and C terminally truncated NR1 subunits. By combining measurements of cell-surface expression of NR2A with co-immunoprecipitation experiments, and by using Blue Native gel electrophoresis to determine the oligomerization status of the subunits, we have shown that regions of the N-terminus of NR1 are critical for subunit association, whereas the truncation of the C-terminus of NR1 before the last transmembrane region has no effect on the association of the subunits. Evidence from the Blue Native gels, sucrose-gradient centrifugation and size exclusion of soluble NR1 domains suggests that NR1 subunits alone can form stable dimers. Using a cell line, which can be induced to express the NMDAR following exposure to dexamethasone, we have shown that NMDARs can be expressed at the cell surface within 5 h of the recombinant gene induction, and that there appears to be a delay between the first appearance of the subunits and their stable association. PMID- 12887324 TI - Assembly and subunit diversity of nicotinic acetylcholine receptors. AB - Nicotinic acetylcholine receptors (nAChRs) are a diverse family of neurotransmitter-gated ion channels which contain five transmembrane subunits arranged around a central pore. Distinct receptor subtypes are expressed at the vertebrate skeletal neuromuscular junction, in mechanosensory cells and within the central and peripheral nervous systems. A total of 17 nAChR subunits (alpha1 alpha10, beta1-beta4, gamma, delta and epsilon ) have been identified in vertebrate species, which can co-assemble to generate a wide variety of nAChRs. Nicotinic receptors also constitute an abundant and diverse family of receptors in invertebrates. As a consequence of studies which have been conducted with both native and recombinant nAChRs, the subunit composition of nAChRs and the rules governing subunit co-assembly are becoming clearer. In this paper the extent of nAChR subunit diversity and evidence for receptor subunit composition is reviewed. PMID- 12887325 TI - Multiple assembly signals in gamma-aminobutyric acid (type A) receptor subunits combine to drive receptor construction and composition. AB - Mammalian gamma-aminobutyric acid type A (GABAA) receptors are constructed from a large repertoire of subunits (alpha1-alpha6, beta1-beta3, gamma1-gamma3, delta, epsilon, theta and pi) into a pentameric ion channel. GABA(A) receptor assembly occurs within the endoplasmic reticulum (ER) and involves interactions with chaperone molecules. Only specific subunit combinations can produce functional surface receptors (with a fixed stoichiometry); other subunit combinations are retained within the ER and degraded. Thus, receptor assembly occurs by defined pathways to limit the diversity of GABAA receptors. The key to understanding how receptor diversity is achieved and controlled is the identification of assembly signals capable of distinguishing between other subunit partners. Analysis of an assembly box in alpha1 (residues 57-68) has revealed an absolute requirement for this region in the assembly of alphabeta receptors. Furthermore, a selective requirement for a single amino acid (R66) is observed for the assembly of alpha1beta2, but not alpha1beta1 or alpha1beta3, receptors. In addition, we have characterized an assembly signal in the beta3 subunit that is capable of driving the assembly of beta3, gamma2beta3 and alpha1beta3 receptors. Interestingly, this signal does not appear to utilize the alpha1 assembly box, suggesting the presence of alternative assembly signals within the alpha1 subunit. Although this beta3 signal is sufficient to permit the formation of betagamma receptors it is not necessary, suggesting that alternative assembly signals also exist within the beta3 subunit. These findings support the belief that GABAA receptor assembly occurs via multiple defined pathways that may be determined by subunit availability. PMID- 12887327 TI - Early events in the trafficking of N-methyl-D-aspartate (NMDA) receptors. AB - The N-methyl-D-aspartate (NMDA) receptor plays a central role at excitatory synapses where it has been implicated in multiple functions associated with synaptic plasticity. While this receptor has been intensely studied with respect to its physiology and pharmacology, its cell-biological properties, such as subunit assembly, post-translational processing and trafficking in neurons, are only beginning to be addressed. Critical to many of the functions of the NMDA receptor are the multiple proteins with which it interacts. While these interactions have been most thoroughly studied with respect to the receptor at the synapse, the same proteins may also interact with the receptor much earlier in its biosynthetic pathway and play important roles in receptor trafficking from the endoplasmic reticulum to the synapse. PMID- 12887326 TI - Real-time imaging of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor (AMPA receptor) movements in neurons. AB - The mechanisms that regulate alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor (AMPAR) synthesis, transport, targeting and surface expression are of fundamental importance for fast excitatory neurotransmission and synaptic plasticity in the mammalian central nervous system. It has become apparent that these control processes involve complex sets of protein-protein interactions and many of the proteins responsible have been identified. We have been working to visualize AMPAR movement in living neurons in order to investigate the effects of blocking protein interactions. Here we outline the approaches used and the results obtained thus far. PMID- 12887328 TI - Pre- and post-synaptic mechanisms regulating the clustering of type A gamma aminobutyric acid receptors (GABAA receptors). AB - Postsynaptic clustering of GABAA (type A gamma-aminobutyric acid) receptors is essential to ensure proper function of GABAergic synapses. This process is initiated during synapse formation and is maintained throughout life. The tubulin associated protein gephyrin is required for clustering of GABAA receptors, but its specific role in this process is not understood. A second protein associated selectively with GABAA receptors at postsynaptic sites is dystrophin. It is present in a subset of GABAergic synapses along with several partners, forming the dystrophin-associated protein complex. In this review, we discuss recent advances in the role of neuronal activity and trans-synaptic signaling for the clustering of gephyrin and dystrophin during synaptogenesis and on the role of these proteins for plasticity and maintenance of mature synapses. PMID- 12887330 TI - Identification of the mitochondrial pyruvate carrier in Saccharomyces cerevisiae. AB - Mitochondrial pyruvate transport is fundamental for metabolism and mediated by a specific inhibitable carrier. We have identified the yeast mitochondrial pyruvate carrier by measuring inhibitor-sensitive pyruvate uptake into mitochondria from 18 different Saccharomyces cerevisiae mutants, each lacking an unattributed member of the mitochondrial carrier family (MCF). Only mitochondria from the YIL006w deletion mutant exhibited no inhibitor-sensitive pyruvate transport, but otherwise behaved normally. YIL006w encodes a 41.9 kDa MCF member with homologous proteins present in both the human and mouse genomes. PMID- 12887329 TI - Hyperthyroidism increases the uncoupled ATPase activity and heat production by the sarcoplasmic reticulum Ca2+-ATPase. AB - The sarcoplasmic reticulum Ca2+-ATPase is able to modulate the distribution of energy released during ATP hydrolysis, so that a portion of energy is used for Ca2+ transport (coupled ATPase activity) and a portion is converted into heat (uncoupled ATPase activity). In this report it is shown that T4 administration to rabbits promotes an increase in the rates of both the uncoupled ATPase activity and heat production in sarcoplasmic reticulum vesicles, and that the degree of activation varies depending on the muscle type used. In white muscles hyperthyroidism promotes a 0.8-fold increase of the uncoupled ATPase activity and in red muscle a 4-fold increase. The yield of vesicles from hyperthyroid muscles is 3-4-fold larger than that obtained from normal muscles; thus the rate of heat production by the Ca2+-ATPase expressed in terms of g of muscle in hyperthyroidism is increased by a factor of 3.6 in white muscles and 12.0 in red muscles. The data presented suggest that the Ca2+-ATPase uncoupled activity may represent one of the heat sources that contributes to the enhanced thermogenesis noted in hyperthyroidism. PMID- 12887331 TI - Caspase processing activates atypical protein kinase C zeta by relieving autoinhibition and destabilizes the protein. AB - Treatment of HeLa cells with tumour necrosis factor alpha (TNFalpha) induced caspase processing of ectopic PKC (protein kinase C) zeta, which converted most of the holoenzyme into the freed kinase domain and increased immune-complex kinase activity. The goal of the present study was to determine the basis for the increased kinase activity that is associated with caspase processing of PKC zeta. Atypical PKC iota is largely identical with PKC zeta, except for a 60-amino-acid segment that lacks the caspase-processing sites of the zeta isoform. Replacement of this segment of PKC zeta with the corresponding segment of PKC iota prevented caspase processing and activation of the kinase function. Processing of purified recombinant PKC zeta by caspase 3 in vitro markedly increased its kinase activity. Caspase processing activated PKC zeta in vitro or intracellularly without increasing the phosphorylation of Thr410 of PKC zeta, which is required for catalytic competency. The freed kinase domain of PKC zeta had a much shorter half-life than the holoenzyme in transfected HeLa cells and in non-transfected kidney epithelial cells. Treatment with TNF-alpha shortened the half-life of the kinase domain protein, and proteasome blockade stabilized the protein. Studies of kinase-domain mutants indicate that a lack of negative charge at Thr410 can shorten the half-life of the freed kinase domain. The present findings indicate that the freed kinase domain has substantially higher kinase activity and a much shorter half-life than the holoenzyme because of accelerated degradation by the ubiquitin-proteasome system. PMID- 12887334 TI - A fragment of the envelope protein from dengue-1 virus, fused in two different sites of the meningococcal P64k protein carrier, induces a functional immune response in mice. AB - Previously we have reported the capacity of the fusion protein PD3, composed of the P64k protein and the envelope (E) fragment from amino acids (aa) 286-426 of dengue-2 virus (DEN-2), to induce a functional immune response in mice against the homologous virus. In that case, the E fragment was inserted within the lipoyl binding domain of the meningococcal P64k protein. In the present study, to test the functionality of the same E region from dengue-1 (DEN-1), a similar construct was made. Furthermore, another alternative of fusion protein was also constructed where the same E fragment from DEN-1 was fused to the C-terminus of the P64k protein. The recombinant proteins obtained (PD11 and PD10) were semi-purified and analysed for their antigenicity, immunogenicity and the ability to protect mice against lethal challenge. Both molecules exhibited the same recognition patterns against anti-DEN-1 polyclonal antibodies. In addition, when administered to mice, they elicited high levels of neutralizing antibodies and induced significant protection against lethal challenge with DEN-1 after intracerebral inoculation. These results reveal the availability of two sites within the P64k for the further insertion of DEN fragments, enabling a construct carrying two fragments from heterologous serotypes within the same molecule of this protein carrier. PMID- 12887335 TI - Influence of sex hormones on the periodontium. AB - OBJECTIVES: Sex hormones have long been considered to play an influential role on periodontal tissues, bone turnover rate, wound healing and periodontal disease progression. The objectives of this review article are to (1) address the link between sex hormones and the periodontium, (2) analyse how these hormones influence the periodontium at different life times and (3) discuss the effects of hormone supplements/replacement on the periodontium. MATERIALS AND METHODS: Two autonomous searches were performed in English language utilizing Medline, Premedline and Pubmed as the online databases. Publications up to 2002 were selected and further reviewed. In addition, a manual search was also performed including specific related journals and books. RESULTS: It is certain that sexual hormones play a key role in periodontal disease progression and wound healing. More specifically, these effects seem to differentiate by gender as well as lifetime period. In addition, the influence of sex hormones can be minimized with good plaque control and with hormone replacement. CONCLUSION: Despite profound research linking periodontal condition with sex hormones kinetics, more definitive molecular mechanisms and therapy still remain to be determined. PMID- 12887332 TI - Free fatty acids induce cell differentiation to infective forms in Trypanosoma cruzi. AB - Intestinal extracts of Triatoma infestans induce cell differentiation of Trypanosoma cruzi epimastigotes into the infective metacyclic form. Part of this effect can be explained by the presence of haemoglobin fragments, which stimulate trypanosomal adenylate cyclase. In this work we examined the metacyclogenic activity of lipids present in this intestinal extract. We found that lipid extracts of the intestinal extract have significant stimulatory effects that reside with the free-fatty-acid fraction, especially oleic acid. These compounds stimulate de novo diacylglycerol formation and protein kinase C activity in the parasite. Moreover, metacyclogenesis is stimulated by phorbol esters and cell permeant diacylglycerol, while protein kinase C down-regulation or incubation with inhibitors of this kinase abrogates this effect. These results indicate that free fatty acids are a novel signal, inducing metacyclogenesis, acting through a pathway involving diacylglycerol biosynthesis and protein kinase C activation. PMID- 12887336 TI - Ultrastructure of the gingiva in cardiac patients treated with or without calcium channel blockers. AB - OBJECTIVES: In the last few years, several studies have suggested that periodontal diseases are related to the development of atherosclerosis and its complications. Our objective was to study the ultrastructural morphology of the gingiva from cardiac patients, some of whom were treated and some not with calcium channel blockers compared to a control group. MATERIAL AND METHODS: Fifty five patients were studied and grouped in the following way: (a) healthy group (HG) (n=12) healthy patients with at least two pockets between 3 and 5 mm; (b) cardiac group (CG) (n=12) patients with cardiac disease untreated with calcium channel blockers; (c) diltiazem group (DG) (n=13) cardiac patients treated with diltiazem; (d) nifedipine group (NG) (n=18) cardiac patients treated with nifedipine. RESULTS: Ultrastructural studies in the CG showed inflammatory cells, collagen fibers disruption and a more extended morphologically compromised fibroblast mitochondria. Morphometric studies in CG showed mitochondria that were impaired in number but increased in volume, suggesting metabolic cell suffering. In DG and NG, morphometric data were similar to HG. The presence of myofibroblasts and collagen neosynthesis was detected in DG and NG. CONCLUSIONS: Our data showed differences in the ultrastructure of the gingival fibroblasts between the studied groups; the DG and NG showed features that could be interpreted as an attempt to restore the cellular metabolic function. PMID- 12887337 TI - Periodontitis and perceived risk for periodontitis in elders with evidence of depression. AB - BACKGROUND: Depression and periodontitis are common conditions in older adults. There is some evidence that these two conditions may be related. AIMS: To study a population of dentate elders and assess the prevalence of depression, self assessment of risk for periodontitis and tooth loss, in relation to periodontal disease status. MATERIAL AND METHODS: Data were obtained from 701 older subjects (mean age 67.2 years (SD+/-4.6), of whom 59.5% were women. Self-reports of a diagnosis of depression, scores of the Geriatric Depression Scale (GDS), and self assessment of risk for future tooth loss and periodontitis were compared with a diagnosis of periodontitis based on probing depth, and bone loss assessed from panoramic radiographs. Other systemic diseases and smoking habits were also determined and studied in relation to depression. RESULTS: A history of depression was reported by 20% of the subjects. GDS scores >/=8 were reported by 9.8% of the elders. Periodontitis was identified in 48.5% of the subjects. Depression was associated with heart attack (p<0.05), stroke (p<0.01), high blood pressure (p<0.02), all combined cardiovascular diseases (p<0.001), chronic pain (p<0.01), osteoarthritis (p<0.001), and osteoporosis (p< 0.001) but not with periodontitis (p=0.73). Subjects with depression had a higher self-reported risk score for future tooth loss (p<0.02). No group difference emerged for self perceived risk for periodontitis. Logistic regression analysis demonstrated that a past history of tooth loss (p<0.001), self-perceived risk for periodontitis (p<0.02), the number of years with a smoking habit (p<0.02), and male gender (p<0.02) were associated with a diagnosis of periodontitis but neither measure of depression could be included in an explanatory model for periodontitis. CONCLUSIONS: Evidence of depression (self-report or by GDS) is not associated with risk for periodontitis in older subjects but is associated with tooth loss and chronic conditions associated with pain. PMID- 12887338 TI - Association between level of education and oral health status in 35-, 50-, 65- and 75-year-olds. AB - AIM: The aim of the present study was to evaluate the association between educational level and dental disease, treatment needs and oral hygiene habits. MATERIAL AND METHODS: Randomized samples of 35-, 50-, 65- and 75-year-olds, classified according to the educational level: [low (LE): elementary school or higher (HE)], were identified. In 1091 subjects, a number of characteristics such as (i) number of teeth, (ii) periodontal attachment levels (PAL), (iii) caries and (iv) occlusal function were recorded. Educational level, oral hygiene and dietary habits were self-reported. Non-parametric variables were analyzed by chi2, Mann-Whitney U-Wilcoxon's rank sum tests, and parametric variables by Student's t-test (level of significance 95%). A two-way anova was performed on decayed, missing and filled surfaces to investigate the interaction between age and educational level. All statistical procedures were performed in the SPSS statistical package. RESULTS: The number of remaining teeth was similar for LE and HE in the 35-year olds (25.8 versus 26.6), but in the older age groups LE had significantly a larger number of missing teeth. The LE groups (except in 65-year olds) exhibited significantly more PAL loss. LE had significantly fewer healthy gingival units in all but the 75-year age group. In all age groups, LE had fewer intact tooth surfaces and a significantly poorer occlusal function. The frequency of tooth cleaning measures and dietary habits did not differ between LE and HE. CONCLUSION: Educational level was shown to influence the oral conditions and should be considered in assessing risk, and in planning appropriate preventive measures. PMID- 12887339 TI - Periodontal repair in dogs: rhBMP-2 significantly enhances bone formation under provisions for guided tissue regeneration. AB - BACKGROUND: Recombinant human bone morphogenetic protein-2 (rhBMP-2) has been shown to support the regeneration of alveolar bone and periodontal attachment in surgically created periodontal defects and in defects with a history of dental plaque and calculus exposure. Periodontal regeneration has also been shown following guided tissue regeneration using space-providing expanded polytetrafluoroethylene (ePTFE) devices. The objective of this study was to evaluate the influence of rhBMP-2 on regeneration of alveolar bone and periodontal attachment used in conjunction with a space-providing ePTFE device. METHODS: Routine, critical-size, 5-6 mm, supra-alveolar, periodontal defects were created around the third and fourth mandibular premolar teeth in four young adult Hound Labrador mongrel dogs. rhBMP-2 (0.2 mg/ml) in an absorbable collagen sponge (rhBMP-2/ACS) or buffer/ACS (control) implants were randomly assigned to be placed around the premolar teeth in the left and right jaw quadrants in subsequent animals. Space-providing ePTFE devices with 300-microm laser-drilled pores, 0.8 mm apart, were used to cover the rhBMP-2 and control implants. The gingival flaps were advanced for primary wound closure. The animals were euthanized at 8 weeks postsurgery for histologic and histometric analyses. RESULTS: Bone regeneration and ankylosis were significantly increased in jaw quadrants receiving rhBMP-2/ACS compared to control (bone height 4.8+/-0.3 versus 2.0+/-0.2 mm, p=0.001; bone area 10.9+/-1.3 versus 1.4+/-0.1 mm2; p=0.009, and ankylosis 2.2+/-0.2 versus 0.04+/-0.7 mm; p=0.01). No differences between groups were found for cementum regeneration and root resorption. CONCLUSIONS: rhBMP-2 significantly enhances regeneration of alveolar bone in conjunction with a space providing, macroporous ePTFE device for GTR. PMID- 12887340 TI - Periodontal repair in dogs: space-providing ePTFE devices increase rhBMP-2/ACS induced bone formation. AB - BACKGROUND: Recombinant human bone morphogenetic protein-2 (rhBMP-2) technologies have been shown to enhance alveolar bone formation significantly. Biomaterial (carrier) limitations, however, have restricted their biologic potential for indications where compressive forces may limit the volume of bone formed. The objective of this proof-of-principle study was to evaluate the potential of a space-providing, macroporous ePTFE device to define rhBMP-2-induced alveolar bone formation using a discriminating onlay defect model. METHODS: Routine, critical size, 5-6 mm, supra-alveolar, periodontal defects were created around the third and fourth mandibular premolar teeth in four young adult Hound Labrador mongrel dogs. All jaw quadrants received rhBMP-2 (0.4 mg) in an absorbable collagen sponge (ACS) carrier. Contralateral jaw quadrants in subsequent animals were randomly assigned to receive additionally the dome-shaped, macroporous ePTFE device over the rhBMP-2/ACS implant or no additional treatment. The gingival flaps were advanced to cover the ePTFE device and teeth, and sutured. Animals were scheduled for euthanasia to provide for histologic observations of healing at 8 weeks postsurgery. RESULTS: Healing was uneventful without device exposures. New bone formation averaged (+/-SD) 4.7+/-0.2 mm (98%) and 4.5+/-0.4 mm (94%) of the defect height, respectively, for jaw quadrants receiving rhBMP-2/ACS with the ePTFE device or rhBMP-2/ACS alone (p>0.05). In contrast, the regenerated bone area was significantly enhanced in jaw quadrants receiving rhBMP-2/ACS with the ePTFE device compared to rhBMP-2/ACS alone (9.3+/-2.7 versus 5.1+/-1.1 mm2; p<0.05). Cementum formation was similar for both treatment groups. Ankylosis compromised periodontal regeneration in all sites. CONCLUSIONS: The results suggest that the novel space-providing, macroporous ePTFE device appears suitable as a template to define rhBMP-2/ACS-induced alveolar bone formation. PMID- 12887341 TI - Efficacy of plaque removal and learning effect of a powered and a manual toothbrush. AB - BACKGROUND: Subjects with high plaque and gingivitis scores can profit most from the introduction of new manual or powered tooth brushes. To improve their hygiene, not only the technical characteristics of new brushes but also the learning effect in efficient handling are of importance. AIM: : The present study compared the efficacy in plaque removal of an electric and a manual toothbrush in a general population and analysed the learning effect in efficient handling. METHOD: Eighty healthy subjects, unfamiliar with electric brushes, were divided into two groups: group 1 used the Philips/Jordan HP 735 powered brush and group 2 used a manual brush, Oral-B40+. Plaque index (PI) and gingival bleeding index (GBI) were assessed at baseline and at weeks 3, 6, 12 and 18. After each evaluation, patients abstained from oral hygiene for 24 h. The next day a 3-min supervised brushing was performed. Before and after this brushing, PI was assessed for the estimation of the individual learning effect. The study was single blinded. RESULTS: Over the 18-week period, PI reduced gradually and statistically significantly (p<0.001) in group 1 from 2.9 (+/-0.38) to 1.5 (+/ 0.24) and in group 2 from 2.9 (+/-0.34) to 2.2 (+/-0.23). From week 3 onwards, the difference between groups was statistically significant (p<0.001). The bleeding index decreased in group 1 from 28% (+/-17%) to 7% (+/-5%) (p<0.001) and in group 2 from 30% (+/-12%) to 12% (+/-6%) (p<0.001). The difference between groups was statistically significant (p<0.001) from week 6 onwards. The learning effect, expressed as the percentage of plaque reduction after 3 min of supervised brushing, was 33% for group 1 and 26% for group 2 at week 0. This percentage increased at week 18 to 64% in group 1 and 44% in group 2 (difference between groups statistically significant: p<0.001). CONCLUSION: The powered brush was significantly more efficient in removing plaque and improving gingival health than the manual brush in the group of subjects unfamiliar with electric brushes. There was also a significant learning effect that was more pronounced with the electric toothbrush. PMID- 12887342 TI - Enamel matrix derivative exhibits angiogenic effect in vitro and in a murine model. AB - OBJECTIVES: Angiogenesis is one of the most critical events in the wound healing process. Any increase in angiogenesis could result in more rapid and complete healing. A recent study found that enamel matrix derivative (EMD) could accelerate early periodontal wound healing. We wanted to clarify whether EMD caused an angiogenic effect and, thus, possibly enhanced wound healing. METHODS: We performed in vitro proliferation and chemotaxis assays on human umbilical vein endothelial cell (HUVEC) cultures, and a tissue culture assay using blood vessel fragments in fibrin gel. Collagen membranes soaked with EMD were implanted subcutaneously in mice to test the in vivo angiogenic effect. RESULTS: While there were no significant differences between the negative control and EMD groups in the proliferation assay, EMD treatment did exhibit a significantly greater dose-dependent chemotactic effect on HUVEC than control group treatments. The tissue culture in fibrin gel showed new blood vessel outgrowths in the EMD groups, but none in the negative control group. In the animal studies, significantly more endothelial cells were detected in the EMD group of mice. CONCLUSIONS: Our findings show that EMD does exhibit some angiogenic effects. However, the underlying molecules and mechanisms are still unidentified. We discuss several possibilities. PMID- 12887343 TI - Evaluation of enamel matrix derivative as an adjunct to non-surgical periodontal therapy. AB - OBJECTIVES: The aim of this study was to evaluate the adjunctive use of enamel matrix derivative (EMD) on periodontal healing following nonsurgical periodontal therapy (scaling and root planing - SRP). MATERIAL AND METHODS: The study was performed as an intraindividual, longitudinal trial of 3 months duration with a double-blinded, split-mouth, controlled, and randomized design. Twenty-two patients with moderate to severe chronic periodontitis were enrolled in the study. In each patient, two sites with pocket depths >/=5 mm and with radiographic angular bone defects >3 mm were selected. Baseline examination included measurement of probing pocket depth (PPD) and clinical attachment levels (CAL). The presence or absence of plaque and bleeding on probing at selected sites was also recorded. Following initial examination, full-mouth SRP was performed. Study sites were then treated with 24% EDTA for 2 min, followed by thorough irrigation with sterile saline. The sites were then randomized. The experimental site received subgingival application of enamel matrix derivative (Emdogain, BIORA AB, Malmo, Sweden). The control site received no additional treatment. At 3 months, all sites were re-examined. The response to therapy in experimental and control sites was evaluated, using change in probing depth and CAL as the primary outcome variables. Statistical analysis (paired t-tests) was used to compare response to treatment in control versus experimental sites. RESULTS: Statistically significant changes in PPD and CAL were seen in both treatment groups from baseline to 3 months. The mean PPD reduction was 2.3+/-0.5 mm for control sites and 2.0+/-0.3 mm for experimental sites. The mean CAL gain was 1.8+/-0.4 mm for control sites, and 1.4+/-0.3 mm for experimental sites. Statistical analysis, however, revealed no significant difference in PPD reduction or CAL gain between experimental and control groups (p>0.4). In addition, no difference was found between treatment groups in bleeding or plaque indices at 3 months. CONCLUSION: The findings from the present study do not support the use of EMD during routine, nonsurgical debridement of periodontal pockets as measured 3 months post SRP. PMID- 12887344 TI - Effectiveness of a combination of platelet-rich plasma, bovine porous bone mineral and guided tissue regeneration in the treatment of mandibular grade II molar furcations in humans. AB - OBJECTIVE: A combination of platelet-rich plasma (PRP), bovine porous bone mineral (BPBM) and guided tissue regeneration (GTR) has been shown to be effective as regenerative treatment for intrabony periodontal defects. The purpose of this study was to evaluate the effectiveness of PRP, BPBM and GTR used in combination as regenerative treatment for grade II molar furcation defects in humans. MATERIAL AND METHODS: Using a split-mouth design, a total of 52 grade II mandibular molar furcation defects were treated either with PRP/BPBM/GTR (experimental group, n=26) or with an open flap debridement (control group, n=26). The primary outcomes evaluated in this study included changes in pocket depth, attachment level and re-entry bone levels (horizontal and vertical) between baseline and 6 months postoperatively. RESULTS: The results showed that the experimental group presented with significantly greater pocket reduction (4.07+/-0.33 mm for experimental and 2.49+/-0.38 mm for control sites), gain in clinical attachment (3.29 +/- 0.42 mm for experimental and 1.68+/-0.31 mm for control sites), vertical defect fill (2.56+/- 0.36 mm for experimental and 0.19+/-0.02 for control sites) and horizontal defect fill (2.28+/-0.33 mm for experimental and 0.08+/-0.02 mm for control sites) than the control group. CONCLUSIONS: It was concluded that the PRP/BPBM/GTR combined technique is an effective modality of regenerative treatment for mandibular grade II furcation defects. Further studies are necessary to elucidate the role played by each component of the combined therapy in achieving these results. PMID- 12887345 TI - Levels of aspartate aminotransferase (AST) in saliva of patients with different periodontal conditions. AB - OBJECTIVES: The purpose of this study was to evaluate the relationship between aspartate aminotransferase (AST) levels in saliva measured by Reflotron trade mark System of Diagnosis and periodontal condition indicated by Community Periodontal Index of Treatment Needs (CPITN). MATERIAL AND METHODS: Fifteen patients were assigned to one of four groups C0, C1, C3 and C4, based on their largest CPITN code among the examined sites, totaling 60 participants. About 1.0 ml of non-stimulated saliva was collected from the individuals after a mouth rinse with water. Biochemical analyses of saliva samples were carried out using the proposed system in order to quantify their AST concentration. RESULTS: There were no significant differences between levels (U/ml) of AST (median; interquartile range) from groups C0 (30.9; 14.7-41.7), C1 (30.3; 19.5-39.4) and C3 (35.1; 27.0-63.5). However, group C4 (106.2; 84.4-129.7) differed statistically from the others (p<0.001) and presented AST levels as high as 284.2 U/ml. Gingival bleeding and suppuration were observed in three individuals with concentrations higher than 125.0 U/ml. CONCLUSION: Levels of AST in saliva from patients presenting CPITN code 4 were higher than from patients coded lower and could be detected by the evaluated diagnostic system. Periodontal destruction such as periodontal pockets, gingival bleeding and suppuration seems to be related to higher AST levels in saliva. PMID- 12887346 TI - Long-term effect of full-mouth tooth extraction on the responsiveness of peripheral blood monocytes. AB - BACKGROUND: As some residual inflammation may remain after periodontal therapy, the present pilot study investigated the long-term effect of full-mouth tooth extraction therapy on the responsiveness of peripheral blood monocytes in a case with generalized terminal adult periodontitis. METHODS: Before and 3, 9, 20 and 32 months after therapy, venous blood was collected. Total and differential white blood cell counts were determined and whole blood cell cultures (WBCC) were incubated with lipopolysaccharide (LPS) to stimulate the production of inflammatory mediators by monocytes. RESULTS: After full-mouth tooth extraction, the numbers of total peripheral white blood cells and neutrophils decreased over time. The release of the chemokines interleukin (IL)-8 and macrophage chemoattractant protein (MCP)-1 in the cultures decreased twofold over time, whereas no changes were seen for the other studied cytokines, chemokines and prostaglandin E2. CONCLUSION: On the basis of previous studies and the present case, the high production of IL-8 and MCP-1 by monocytes in LPS-stimulated WBCC from periodontitis patients is most likely acquired, as their levels decrease over time when the periodontal infection is controlled. The possible connection between periodontitis and atherosclerosis through IL-8 and MCP-1 is discussed. PMID- 12887347 TI - What's the scoop on the nursing shortage? PMID- 12887349 TI - The nursing shortage in the United States of America: an integrative review of the literature. AB - AIM: The aims of this paper are to review the literature to determine what factors are contributing to the nursing shortage in the United States of America (USA) and discuss possible solutions to this current and future nursing shortage. BACKGROUND: The need for nurses is often depicted as cyclical in nature. Throughout history, the USA has experienced a series of nursing surpluses and shortages. However, the current shortage has been characterized as being unlike those experienced in the past. Trends of an ageing Registered Nurse (RN) workforce and limited supply to fill the impending vacancies are some of the unique aspects that bring a new dimension to an old problem. Today's nursing shortage will not be resolved by simply returning to the solutions of yesteryear, and strategies to reduce its impact will have to be more creative and focus on the long-term. METHODS: Integrative literature review of published literature on the current nursing shortage in the USA from 1999 to 2001. DISCUSSION: Four main areas were identified as the major contributors to the nursing shortage in the USA: the ageing RN workforce; declining enrollment; the changing work climate; and the poor image of nursing. Solutions to the shortage followed similar themes to the contributing factors and encompassed four main areas: exploring recruitment efforts; exploring retainment efforts; improving the image of nursing; and supporting legislation that helps to rectify the shortage. CONCLUSION: There is firm evidence that the USA amidst a nursing shortage. Much is known about the many contributing factors but now nurses need to become proactive to help secure the future of their workforce. By forming partnerships within the profession and with other influential parties, nurses can be in the forefront of resolving their workforce issues. PMID- 12887350 TI - Commentary: an Australian perspective. PMID- 12887351 TI - Commentary: an English perspective. PMID- 12887352 TI - Commentary: a European perspective. PMID- 12887353 TI - Reconstructing mental health nursing in home care. AB - BACKGROUND: Psychiatric care for people in Sweden with long-term mental health problems has shifted from institutional to community and home settings. AIM: The aim of this study was to explore and describe mental health nurses' experiences of how structural changes in mental health nursing influence interaction when providing home care to patients with long-term mental health problems. METHOD: We conducted interviews with 11 mental health nurses who provided home care to people with long-term mental health problems. The constant comparative method of grounded theory was used for data collection and analysis. FINDINGS: The process of attempting to reconstruct mental health nursing by entering into the everyday reality of people with long-term mental health problems was identified as the core category. Central to this process was a change in perspective from nurse controlled to client-centred care. This led to changes in moral values, assessment of client needs, and ways of meeting clients and establishing relationships. However, attempts to reconstruct mental health nursing according to a client-centred perspective became problematic when clients were at risk of harming themselves. In these situations, respondents felt the need to make decisions for clients, and this was experienced as burdensome and lonely. CONCLUSIONS: Our study indicates that the process of reconstructing mental health nursing in line with a client-centred perspective is incomplete. Opportunities to interact with people with mental health problems in the home environment may lead to a broader perspective on the daily lives of these people, but also to a realization that home care cannot be modelled on institutional care. PMID- 12887354 TI - Eating difficulties after stroke. AB - BACKGROUND: Stroke is a common and enduring problem, producing a wide range of effects that may impact on all aspects of life. One area that has seldom been investigated is the effects of stroke on ability to eat and, particularly, stroke survivors' subjective experiences of eating-related difficulties. AIM: To investigate stroke survivors' reports of eating-related experiences 6 months after stroke. METHODS: Participants were 206 survivors of acute stroke who were admitted to hospital between March 1998 and April 1999. Those able to communicate and who gave informed consent participated in a semi-structured interview and assessment of eating abilities in their homes 6 months after their stroke. Interviews were tape recorded and transcribed and 113 interviews with eating disabled participants were entered onto QSR NUD*IST 4 for thematic analysis. Analyses were later checked independently. FINDINGS: At 6 months, 34%, 61% and 5% had no, slight and moderate eating disablements, respectively. A range and variety of difficulties were discussed, but relationships between degrees of disablement and handicap were not straightforward; effects seemed more closely related to participants' responses than objective difficulties. Eating and related activities were clearly important aspects of life for these stroke survivors, socially and psychologically, as well as functionally. Prestroke activities were sometimes maintained, with considerable effort. CONCLUSIONS: Findings indicated issues of relevance for those involved with early rehabilitation interventions and highlighted aspects of continuing care service delivery that warrant review. The relative lack of attention paid to eating related aspects of care is an important oversight. PMID- 12887355 TI - Self-efficacy training for patients with end-stage renal disease. AB - BACKGROUND: Patients with end-stage renal disease often fail to follow a prescribed diet and fluid regimen, which undermines the effectiveness of care and leads to unpredictable progression of the disease and greater likelihood of complications. AIM: The purpose of this randomized controlled study was to examine the effectiveness of self-efficacy training on fluid intake compliance. METHODS: This study took place in northern Taiwan. Eligible patients were receiving routine haemodialysis; 20-65 years of age; living in a community setting; able to read and write; and willing to participate. Sixty-two end-stage renal disease patients participated in the study. Those in the experimental group (n = 31) received 12 sessions of structured self-efficacy training; the control group patients (n = 31) received only routine care. The intervention was based on Bandura's theory and included an educational component, performance mastery, experience sharing, and stress management. The outcome measure was the mean body weight gain between dialysis sessions. Data were collected at baseline, 1, 3 and 6 months following the intervention and analysed by a descriptive and repeated measures anova. RESULTS: Experimental group mean weight gains decreased gradually following self-efficacy training. However, control group mean weight gains decreased only slightly over time. These results were statistically significant when baseline differences controlled for (P < 0.05). CONCLUSIONS: The study supports the effectiveness of the self-efficacy training in controlling mean body weight gains of end-stage renal disease patients receiving haemodialysis. PMID- 12887356 TI - Patient on display--a study of everyday practice in intensive care. AB - BACKGROUND: This study investigates the situated organization in a workplace producing intensive care, that is an intensive care unit (ICU). The workplace research tradition concerns work and interaction/communication in technology intensive environments. Communication is seen as social action and cannot be separated from production or from the context in which the activities are situated. AIM: The aim of the present study was to explore how intensive care is produced by analysing a recurrent situated activity in the ICU, namely the delivery and reception of a patient coming from the operation unit. METHOD: In the fieldwork, participant observations was used to study everyday practice in an ICU, combined with written field notes. FINDINGS AND DISCUSSION: Intensive care is to a great extent produced through routine practices. The division of labour is marked and is taken for granted: everyone knows what to do. The actors' physical location in the room is connected to their functions and work with supportive tools. Verbal reports, visual displays and activities make the information transmission available to everyone in the patient room. Shared understanding of the situation seems to make words redundant when the activities of competent actors are co-ordinated. There is also coordination between the actors in the ICU and the technological equipment, which constantly produces new information that must be interpreted. Enrolled Nurses are physically closest to the patients, the physician is the one most physically distant from patients and Registered Nurses bridge the gap between them. These actors produce and re produce intensive care through constant sense-making in the here and now at the same time as the past is present in their activities. PMID- 12887357 TI - Effects of episiotomy on bonding and mothers' health. AB - BACKGROUND: Historically, episiotomy has been carried out during labour to facilitate delivery, shorten the duration of the second stage and prevent spontaneous lacerations. However, recent studies of episiotomy have recommended that it be carried out only when necessary. In Turkey, midwives are authorized to perform episiotomy. AIM: To analyse the effects of episiotomy on mothers' health and mother-infant bonding. DESIGN: A cohort study of 100 women who gave birth by normal vaginal delivery in a Turkish hospital between 15 March 1999 and 6 April 2000. METHODS: Participants were divided into episiotomy (n = 50) and control (n = 50) groups. Data on biographical characteristics and the process of labour were collected in hospital, and follow-up was conducted at home 1, 3 and 12 weeks after labour. RESULTS: Mean duration of the second stage was longer in the episiotomy group than the control group, but the difference was not significant. The number of spontaneous lacerations was significantly lower in the episiotomy group. Mean time from delivery to maternal rest and time taken to bond with the infant were significantly longer in the episiotomy group. There were significantly higher scores for overall incidence and severity of pain on the first day, and pain incidence and severity at 1 week in the episiotomy group. Significantly fewer women in the episiotomy group were able to do chores and to sit/stand up comfortably in the first postpartum week CONCLUSION: Episiotomy should not be used unless indicated. Measures should be taken to avoid perineal trauma during labour, establish bonding between mother and infant as soon as possible, and minimize perineal discomfort after delivery. PMID- 12887358 TI - Bullying in the nursing profession. AB - BACKGROUND: Self-esteem is a major predictor of behaviour. Nurses with healthy self-esteem are likely to deliver therapeutic patient care, while those with low self-esteem are less likely to do so. AIM: The aim of the 3-year study discussed here was to explore students' self-esteem and how their experiences of preregistration education influenced its development over the period of the programme. RESEARCH METHODS: Students participated in unstructured qualitative interviews at the beginning and end of their 3-year preregistration course and a grounded theory approach was used for data collection and analysis. FINDINGS: Bullying was found to be commonplace in the transition to becoming a nurse. Students were bullied and also witnessed patients being bullied by qualified nurses. The internalization of nursing norms meant that students then bullied others. Students' self-esteem was low. CONCLUSION: Bullying, and its effects on self-esteem, are perpetuated by practices within nursing. This situation will only be changed if nurses and educators transform their practice and the context in which bullying occurs. Otherwise, each new generation of nurses will continue to be socialized into negative practices which undermine both their own feelings of self-worth and standards of nursing care. PMID- 12887359 TI - Experiences of using the VIPS-model for nursing documentation: a focus group study [corrected]. AB - BACKGROUND: The present investigation is part of a study where the Registered Nurses on three hospital wards received a 2 year intervention programme on nursing documentation in accordance with a keyword structure based on the nursing process. AIM: To describe the Registered Nurses' perceptions of and attitudes towards the effects of the intervention, and to generate hypotheses for further research. METHOD: Focus group discussions were used to collect data, with a qualitative content analysis method for the processing of the data. FINDINGS: The most interesting finding in these group discussions was the statements made by participants that the structured way of documenting nursing care made them think more, and think in a different way about their work with their patients. Two types of role changing were reported; from a medical technical focus to a more nursing expertise orientation and from a "hands on clinician" to more of an administrator and secretary. CONCLUSION: A number of issues debated among the participants in this study could be seen as organizational matters and lead to the important issue of multidisciplinary and organizational work when implementing innovations within nursing. PMID- 12887360 TI - Critical realism: a way forward for evaluation research in nursing? AB - BACKGROUND: Critical realism is a relatively new philosophical perspective that combines a realist ontology with a relativist epistemology in subscribing to a form of "robust" relativism. The potential benefits of adopting a critical realist approach are considered in two areas of evaluation research: theory driven programme evaluation and policy evaluation. AIM: The aim of the paper is to identify the main features of critical realism and explore its potential as a philosophical framework in the field of evaluation research. CONCLUSION: We argue that critical realism provides a coherent framework for evaluation research that is based on the understanding of causal mechanisms. This is particularly important for frontline services seeking to use evidence-based interventions, as interventions need to be properly understood if they are to be used effectively in the context of clinical practice. Critical realism can also add depth to policy analysis. However, there is a potential for critical realists to drift beyond the boundary that separates scientific knowledge from speculative theory when making recommendations about the best course of action to follow. PMID- 12887361 TI - Urolithiasis in Okinawa, Japan: a relatively high prevalence of uric acid stones. AB - AIM: The aim of the present study was to investigate the composition of urinary tract stones in patients from Okinawa, the most southern island group of Japan. METHODS: The study was conducted by 12 hospitals in Okinawa. A total of 1816 urinary tract calculi were obtained from 1816 patients (1323 males; 493 females). The patients had a mean age of 53 +/- 15.3 years (mean +/- SD). The calculi were examined to determine their chemical composition. Stone samples were analyzed by computed infrared spectrophotometer. RESULTS: Pure stones comprised 58.4% of the total, with calcium oxalate stones accounting for 40% (21% monohydrate [whewellite]; 6.6% dihydrate [weddellite]; and 12.4% combined monohydrate and dihydrate stones), uric acid/urate stones for 9.6%, calcium phosphate stones for 5.1%, and struvite stones for 3.7%. The other 41.6% of the stones consisted of calcium oxalate mixed with other components. The male-to-female ratio was 2.7:1. CONCLUSION: In our series, calcium oxalate stones accounted for 81.6% of the urinary tract calculi, while uric acid/urate stones accounted for 15.8%. Uric acid stones, predominantly the anhydrous and/or dihydrate forms, showed a relatively high prevalence. Calcium oxalate stones, predominantly in the form of whewellite, showed a high prevalence among pure calculi; while the predominant combinations among mixed calculi were weddellite + calcium phosphate and whewellite + uric acid/urate. PMID- 12887362 TI - Device to promote pelvic floor muscle training for stress incontinence. AB - AIM: Many patients with stress urinary incontinence do not have enough motivation to continue pelvic floor muscle training (PFMT) by themselves. Therefore, a device was created to support PFMT, and its effect was examined. METHODS: Forty six women with stress urinary incontinence were assigned to a control group or a device group in order of presentation. A pamphlet on PFMT was given to control patients, while the same pamphlet plus the device and instructions on its use were given to patients in the device group. The device had a chime that was set to sound three times a day when exercise sessions were scheduled. PFMT consisted of fast and slow pelvic floor muscle contraction exercises that were performed for 2 min and followed a rhythm set by the device. RESULTS: After 8 weeks, 20 patients from the control group and 21 patients from the device group could be evaluated. In the control group, only the quality of life (QOL) index improved significantly. In the device group, however, the daily number of incontinence episodes, the number of pads used daily, the QOL index, and the pad weight in the pad test improved significantly. Patients in the device group said that they felt obligated to perform PFMT when the chime sounded. Forty-eight percent of patients from the device group were satisfied with the outcome of PFMT, while only 15% were satisfied in the control group. CONCLUSION: This device may be useful to support the management of stress urinary incontinence. PMID- 12887363 TI - Determinants of quality of life of bladder cancer survivors five years after treatment in France. AB - OBJECTIVES: The aim of the population-based study presented here was to evaluate quality of life (QOL) among bladder cancer survivors at least five years after diagnosis, ascertain the long term effects of treatments, and investigate important pathological and sociodemographic factors influencing the QOL of such survivors. METHODS: A Functional Assessment of Cancer Therapy (FACT) questionnaire with a bladder-specific option (FACT-Bl), was sent by mail to patients randomly selected from the database of the Isere and Tarn cancer registries in France. Ninety-five valid completed questionnaires were returned by 78 males and 17 females, with a median age of 72 years (range, 33-90). The pathological information contained in the completed questionnaires was examined. RESULTS: Of the 95 patients (76 with a superficial tumor, 17 with an invasive tumor), 20 had undergone total cystectomy. Total cystectomy clearly deteriorates the autonomy of survivors (ECOG-PSR), as well as their scores on the bladder cancer-specific sub-scale in sexuality-related questions. Survivors claimed impotency and loss of sexual interest after the cystectomy. In contrast, neither the type of treatment nor the time from the most recent major treatment were score-modifying factors. Negative psychological effects of treatments in the long term were not observed. Rather, patients' QOL was affected by their autonomy in daily life and by old age. In addition, familial situation had a critical impact on emotional and Familial/Social Well-being. CONCLUSION: The present study showed the long-term negative effect of total cystectomy on survivors. Survivor autonomy and other sociodemographic backgrounds were also determinants of their QOL. The maintenance of good health, sexual function and active family relationships should be promoted so that patients can lead full lives after treatment. PMID- 12887364 TI - Morbidity of laparoscopic radical prostatectomy: summary of early multi institutional experience in Japan. AB - AIM: Laparoscopic radical prostatectomy is being evaluated throughout the world. The aim of the present study is to report early multi-institutional experience of the procedure in Japan. METHODS: A total of 148 men who were diagnosed with clinically localized prostate cancer underwent laparoscopic radical prostatectomy at seven different institutions in Japan. Early complications (within 30 days postoperatively) and postoperative convalescence were reviewed retrospectively. The median age of patients was 68.0 years (range, 51-80). RESULTS: The median operative time was 403 minutes (range, 167-925; average, 427). Blood loss ranged from 50 to 5000 mL (median, 540; average, 856). A total of 66 complications were reported in 55 patients (37.2%). Intraoperative complications were noted in 25 of 148 patients (16.9%): 10 rectal injuries (6.8%); five bladder injuries (3.4%); five cases of subcutaneous emphysema (3.4%); two intestinal injuries (1.4%); one major vessel injury (0.7%); one ureteral injury (0.7%); and one obturator nerve injury (0.7%). Overall, 16 of 148 patients (10.8%) required open conversion or postoperative open surgical repair. The most common postoperative complications were anastomotic leakage (6.8%), wound-related complications (4.7%) and perineal pain (4.7%). The bladder catheter was removed on day 7 or earlier in 73 cases (49.3%). The median time to ambulation was 1 day (mean 1.4, range 1-5). Oral intake was started on postoperative day 1 in 67 patients (45.2%) and on postoperative day 2 in 65 (43.9%). CONCLUSION: Although laparoscopic radical prostatectomy is technically demanding, reduced blood loss and shorter convalescence periods can be expected from the procedure. Surgeons should be aware of the disturbingly high morbidity rate related to early experience. By mastering laparoscopic skills and sharing knowledge, surgeons could reduce the impact of the learning curve required to complete this procedure competently. PMID- 12887365 TI - Retrospective analysis of 74 cases of seminoma treated with radiotherapy. AB - BACKGROUND: Standard post-orchiectomy radiotherapy (RT) is accepted as a standard management option for stage I seminoma. METHODS: Retrospective evaluation of 74 patients with stage I seminoma was performed according to the Royal Marsden staging system. All of the patients underwent RT in the Radiation Oncology Department of Gulhane Military Medical Academy between 1974 and 1995. The median age of patients was 27 years (range, 20-56). Radiotherapy was applied to all of the patients after orchiectomy for adjuvant purposes. Sixty-nine patients underwent RT while five patients who had recurrence received chemotherapy following radiotherapy. RESULTS: After a mean follow-up period of 54 months, the 5-year overall survival rate was 98.61%, which complied with the literature. The disease-free survival rate was 90.54%. According to the World Health Organization toxicity scale, acute enteritis was 9.4% for grade I and 5.4% for grade II, while nausea/vomiting was 36.4% for grade I and 5.4% for grade II. CONCLUSION: To avoid acute toxicity related to RT, prognostic risk factors should be well-known and patients with low risk factors should be monitored carefully after orchiectomy. RT should be directed to the para-aortic +/- ipsilateral pelvic lymph nodes in high risk patients. Although post-orchiectomy RT is a traditional management option for clinical stage I seminoma, the results of RT should be well-known to compare it with other treatment options (e.g. RPLND, adjuvant chemotherapy and surveillance). PMID- 12887366 TI - Further investigation of the epitope recognized by the new monoclonal antibody 2C9. AB - OBJECTIVE: We established a new monoclonal antibody (2C9) that reacted with prostate tissue. The immunohistochemical reactivity of this antibody is similar to anti-prostate-specific membrane antigen (PSMA). Herein, we report the antigenic determinant of 2C9 antibody. METHODS: The reactivity of the antibody was characterized by immunohistochemical staining and the antigen target was characterized by amino acid sequencing after immuno-affinity purification from an LNCaP cell lysate and cloning of a cDNA using a mammalian expression cDNA cloning system. RESULTS: The amino acid and nucleotide sequences for the antigen molecule recognized with 2C9 monoclonal antibody demonstrated identity with PSMA. CONCLUSION: The target molecule of the 2C9 monoclonal antibody is PSMA, pointing to future diagnostic and therapeutic applications. PMID- 12887367 TI - Successful long-term disease-free survival following multimodal treatments in a patient with a repeatedly recurrent refractory adrenal cortical carcinoma. AB - A 47-year-old male patient underwent surgery for a 10-cm adrenal cortical carcinoma. A large invasive adrenal mass was surgically removed en bloc with the right kidney and the lower lobe of the liver. Two months postoperatively, a 7-cm recurrent mass developed in the right psoas muscle. After a partial response was achieved by irradiation (40 Gy) and high-dose chemotherapy (carboplatin and etoposide) with peripheral blood stem cell transplantation, the patient underwent surgery with a wide excision of the psoas muscle. Twelve months after the initial surgery, an 8-cm rib metastasis developed and the patient again underwent surgery after a combination of irradiation (50 Gy) and chemotherapy (cisplatin and etoposide). The patient has been doing well without any evidence of recurrence for 5 years. Refractory or metastatic adrenal cortical carcinomas have been thought to be lethal, therefore, the present case provides support for multimodal treatments of refractory adrenocortical cancers. PMID- 12887368 TI - Adrenocorticotropin-independent macronodular adrenal hyperplasia treated by simultaneous bilateral laparoscopic adrenalectomy. AB - A 61-year-old woman was admitted to our hospital with hypertension and diabetes mellitus, and was found to have Cushing's syndrome. Radiological and endocrinological findings suggested adrenocorticotropic hormone-independent macronodular adrenal hyperplasia. Simultaneous bilateral laparoscopic adrenalectomy was performed, minimizing the number of trocar sites and operation time. Success was attributed to the careful selection of trocar sites to permit safe dissection. PMID- 12887369 TI - Metastatic renal cell carcinoma to the bladder 12 years after radical nephrectomy. AB - We report, herein, a case of metastatic renal cell carcinoma of the urinary bladder. A 76-year-old man presented to our hospital. He had undergone right radical nephrectomy at 64 years of age. Cystoscopy revealed a solitary, spherical tumor 1.5 cm in size protruding into the urinary bladder. Transurethral resection was performed and the pathological diagnosis of the lesion was clear cell carcinoma. The patient is alive 12 months after recurrence to the bladder, under the administration of interleukin-2. PMID- 12887370 TI - Signet-ring cell carcinoma of the prostate effectively treated with maximal androgen blockade. AB - Primary signet-ring cell carcinoma (SRCC) of the prostate is very rare and has a poor prognosis, even when treated with aggressive therapy. We report herein a case of a 72-year-old man with prostatic SRCC. The patient had a tumor that extended directly to the rectum. Maximal androgen blockade was started and 20 months later, the patient was alive without evidence of recurrence. The present case of prostatic SRCC responded well to medical therapy, however, tumors can recur after a long period of time. Therefore, adjuvant therapy is recommended. PMID- 12887371 TI - Papillary adenoma of the prostatic urethra: report of two cases. AB - Papillary adenoma of the prostatic urethra is a rare cause of hematuria, hemospermia and lower urinary tract obstruction, especially in young adults. We report two cases of papillary adenoma and their endoscopic views. One of the patients had a lesion located on the verumontanum and presented with hematuria, hemospermia and symptoms of lower urinary tract obstruction. The other patient had papillary lesions located in the paramontanal gutter and presented with persistent painless hematuria. The origin of both lesions was proven to be prostatic epithelial differentiation using immunohistochemical identification of prostate-specific antigen. Papillary adenoma is believed to be benign, therefore, both lesions were carefully excised and fulgurated transurethrally. PMID- 12887372 TI - Support to heal the scars of war. PMID- 12887373 TI - Leading the development of nursing within a Nursing Development Unit: the perspectives of leadership by the team leader and a professor of nursing. AB - Leadership within nursing is receiving unprecedented focus and development. This reflective narrative explores the nature of leadership, termed scholarly leadership, by an academic and a clinical leader of a Nursing Development Unit. The narrative explores the characteristics of such leadership and highlights how it empowered a nursing team to further reach its potential. Two areas, patient centered care and the characteristics of practice, are focused upon to highlight the leadership style that the clinical leader adopted. The paper concludes by suggesting what structural and systems changes need to be put in place in order to bring about change. PMID- 12887374 TI - Hospital fall risk assessment tools: a critique of the literature. AB - There are numerous fall risk assessment tools developed for use by nurses in hospitals; however, few of them have established validity and reliability. This article will examine the current state of knowledge in regard to fall risk assessment tools through review and critique of the literature on the topic. Recommendations for future research on fall risk assessment tools will be made and a conceptual framework detailing the relationship between the variables involved in assessing the accuracy of fall risk assessment tools will be presented. PMID- 12887375 TI - The influences on and experiences of becoming nurse entrepreneurs: a Delphi study. AB - There is little known about private practice nursing as an area of advanced practice. As more nurses take the option to develop private practice, the experiences of and influences on nurses currently in private practice might be a useful guide to the pitfalls and difficulties which might be encountered. In addition, an understanding of the experiences of and influences might assist nursing organizations and health services to provide support to nurses who play an integral part in health care delivery in the community. A research study was undertaken utilizing a two-round Delphi Technique to elicit and assess consensus on the reasons for nurses going into business and the experiences they encountered in becoming and being a nurse entrepreneur. The study instrument in round one comprised a questionnaire with statement headings inviting opinions on the influences and experiences of nurses in business. In the second round, levels of agreement were elicited from responders on collated opinions from round one, including statements formed from comments written in round one. The initial questionnaire also included closed questions to obtain a profile of nurses in private practice. Responders were 59 nurses in private practice in round one and 54 nurses in round two. The themes raised could be grouped under headings of influences, advantages/disadvantages, education/experience, skills/knowledge, characteristics and barriers. The level of agreement on the themes was reasonably high. Dissent occurred on issues of increased income, professional image and support structures. This Delphi study has identified key areas of consensus on the experiences of nurses in private practice who have extended their career into the business arena. It has also identified areas in which further work needs to be carried out to understand this work of nurse entrepreneurs. PMID- 12887376 TI - Improvement of family carers' knowledge, skills and attitudes in caring for older people following the implementation of a Family Carers' Training Program in the community in Indonesia. AB - The purpose of this study was to examine family carers' knowledge, skills and attitudes in caring for older people following the implementation of a Family Carers' Training Program (FCTP). Family carers (n = 240) from two villages in Indonesia were randomly selected. All participants were surveyed by questionnaires and their care behaviour observed at baseline, three and six months. Findings indicated significant increases in knowledge, skills and attitudes in the intervention group over six months compared to those of the control group. Hence, the FCTP demonstrated a positive effect on family carers' capabilities and attitudes in caring for older people living in the community in the study setting. PMID- 12887377 TI - Consumer participation in acute care settings: an Australian experience. AB - Consumer partnerships have been embraced as an important component of building high quality health care services. While nurses have the greatest contact with clients in hospital, little is known of their views about consumer participation or how they facilitate that participation at the bedside. Using focus group interviews and participant observation methods, this project explored nurses' approaches to working with consumers to support their participation in health care delivery. Findings indicate a sharp contrast between the ideas that nurses expressed and the actions observed in practice. It was clear from the interviews that nurses had adopted the rhetoric surrounding consumer participation, yet observational data revealed nursing practices that excluded active participation by consumers. Factors influencing nurses' facilitation of consumer participation were identified as the division of nursing labour in the setting, limited communication between nurses and patients and environmental constraints. PMID- 12887378 TI - Workplace violence in nursing in Queensland, Australia: a self-reported study. AB - This paper reports on workplace violence, self-reported as part of a survey among members of the Queensland Nurses' Union (QNU) in October 2001. The aim of the overall study was to ascertain how nurses perceived their work and their working conditions and to use the results of the study to inform strategic planning of the QNU. The participants of this study were drawn from a stratified random sample of 2800 QNU members employed in the public, the private acute and the aged care sectors. The total number of completed surveys analysed was 1436: 441 aged care, 497 public and 498 private acute sector surveys. The results suggest that the rate of workplace violence differs significantly across sectors. In the three months immediately prior to the survey, 50% of aged care sector nurses experienced some form of workplace violence compared to 47% of public sector nurses and 29% of acute private sector nurses. The major source of workplace violence was from patients. The second most common source of violence varied. Nurses in the public sector signified visitors and relatives while nurses in the aged care and acute private sectors indicated other nurses. The study found that inexperienced nurses are more likely to report workplace violence than experienced nurses. Additionally, the more experienced the nurse, the less likely they are to perceive workplace policies and procedures for workplace violence as effective. Additional findings include an apparent lack of policies and procedures for workplace violence in rural and remote areas, and an association between the designation of a nurse and the reported level of workplace violence. PMID- 12887379 TI - Incidence of patent furcal accessory canals in permanent molars of a Turkish population. AB - AIM: To determine the incidence of patent furcal accessory canals in permanent molars of a Turkish population. METHODOLOGY: Two-hundred extracted teeth consisting of 50 maxillary first molars, 50 maxillary second molars, 50 mandibular first molars and 50 mandibular second molars from Turkish patients attending the Oral Surgery Department of the Faculty of Dentistry, Istanbul University, Istanbul, Turkey, were included in the study. After preparation of access cavities and removal of pulp tissue, the teeth were stored in 5.25% sodium hypochlorite for 1 h. Following double-sealing of the access cavities, all tooth surfaces except the furcation regions were covered with nail varnish. The teeth were stored in 0.5% basic fuchsin for 1 week. The teeth were sectioned at the cemento-enamel junction and the presence of patent furcal canals was established by examining the pulp chamber floor with a stereomicroscope (10x) to determine staining. RESULTS: Patent furcal accessory canals were detected in 24% of maxillary first molars, 16% of maxillary second molars, 24% of mandibular first molars, and 20% of mandibular second molars. No statistically significant differences were found between the tooth types. CONCLUSIONS: In a Turkish population, the incidence of patent furcal accessory canals on the pulp chamber floor of maxillary and mandibular first and second molars ranged between 16 and 24%. PMID- 12887380 TI - A prospective clinical study of Mineral Trioxide Aggregate and IRM when used as root-end filling materials in endodontic surgery. AB - AIM: To assess the success rate of the root-end filling material, Mineral Trioxide Aggregate (MTA). METHODOLOGY: Referred adult patients were recruited using strict entry criteria and randomly allocated to receive MTA or IRM. A standardized surgical technique was employed: the root end was resected perpendicularly and a root-end cavity was prepared ultrasonically and filled. A radiograph taken immediately after surgery was compared with those taken at 12 and 24 months. Customised film holders and the paralleling technique were used; radiographs were assessed by two trained observers using agreed criteria. The results from 122 patients (58 in IRM group, 64 in MTA group) after 12 months and 108 patients (47 in IRM group, 61 in MTA group) for the 24-month review period were analysed using the chi2 test. RESULTS: The highest number of teeth with complete healing at both times was observed when MTA was used. When the numbers of teeth with complete and incomplete (scar) healing, and those with uncertain and unsatisfactory healing were combined, the success rate for MTA was higher (84% after 12 months, 92% after 24 months) compared with IRM (76% after 12 months, 87% after 24 months). However, statistical analysis showed no significant difference in success between materials (P > 0.05) at both 12 and 24 months. CONCLUSIONS: In this study, the use of MTA as a root-end filling material resulted in a high success rate that was not significantly better than that obtained using IRM. PMID- 12887381 TI - Rheological properties and biocompatibility of endodontic sealers. AB - AIM: To determine the flow characteristics and subcutaneous tissue reactions to five endodontic sealers. METHODOLOGY: The materials used were Procosol, AH26, Endomethasone, Sealapex and Endion. The sealers were prepared following the manufacturers' instructions, and 0.075 mL of each material was placed on a glass surface, which was then rotated 90 degrees. The samples were stored at 37 degrees C and 95% humidity. The displacement of the sealer was recorded by measuring the difference between its original position and the position recorded at 15 and 60 min. Three samples of each material were used. Two pockets were created in the back of Wistar rats, and one silicone tube, 1 mm in diameter and 1 cm in length, was implanted in each. One was filled with one of the materials under study, and the other empty tube was implanted as a control. Fourteen days after implantation, the animals were sacrificed, and samples of the skin containing the tubes were histologically processed. Histological and histomorphometric evaluations of the tissues adjacent to the open end of the tube were carried out the volume of tissue reaction was measured histomorphometrically according to standard stereological principles. Results were statistically analysed using analysis of variance and Duncan's test. RESULTS: The highest flow values were obtained with Sealapex and AH26. Time significantly affected the flow and the material (P < (1001). Procosol and Endion produced the most severe histological reactions: these were outlined by fibrous tissue; AH26. Endomethasone and Sealapex produced reactions of smaller size and with more moderately defined limits. CONCLUSIONS: The flow did not correlate with the degree of inflammatory response. Procosol and Endion produced the most severe tissue reactions, whereas Endomethasone, Sealapex and AH26 produced only minimum reactions. PMID- 12887382 TI - Fluid movement along the coronal two-thirds of root fillings placed by three different gutta-percha techniques. AB - AIM: To compare fluid movement (FM) along the coronal two-thirds of gutta percha/sealer root fillings placed by three different techniques. METHODOLOGY: Three groups consisting of the roots of mandibular incisors were instrumented and obturated by cold lateral compaction (LC), warm vertical compaction (VC) or the single-cone technique (SC), using RoekoSeal Automix (RSA) as the sealer. The apical 4 mm of each root filling was removed, and FM along the remaining 7 mm of the root filling was measured. RESULTS: The VC group displayed more FM than the other two groups (P = 0.023). No significant difference in FM was found between the LC and SC groups (P = 0.629). CONCLUSIONS: The coronal two-thirds of the VC root fillings did not prevent FM when RSA sealer was used. PMID- 12887383 TI - The location of the cementodentinal junction in single-rooted mandibular first premolars from Egyptian and Saudi patients: a histological study. AB - AIM: To determine the location of the cementodentinal junction (CDJ) in mandibular first premolars from Egyptian and Saudi patients. METHODOLOGY: One hundred and twenty-two extracted single-rooted mandibular first premolars (68 Egyptian; 54 Saudi) were examined histologically. Half of the teeth were fixed, decalcified, embedded in paraffin, mesio-distally sectioned and stained with H&E, whilst the other half were ground with a stone prior to microscopic evaluation. Measurements of the location of the CDJ were made parallel to the long axis of the teeth from the external surface of the root apex on the mesial and distal sides of the canal. Measurements were statistically analysed using two-way anova of repeated-measure design. RESULTS: The CDJ in Egyptian sections ranged from 0 to 2.0 mm (mean +/- SD = 0.9 +/- 0.4 mm) on the mesial side of the canal and from 0.2 to 2.5 mm (mean +/- SD = 1.0 +/- 0.5 mm) on the distal aspect. In the Saudi sample, the CDJ ranged from 0.5 to 1.2 mm (mean +/- SD = 0.8 +/- 0.3 mm) on the mesial side and from 0.6 to 0.9 mm (mean +/- SD = 0.7 +/- 0.4 mm) on the distal side. The mesial and distal differences between Egyptian and Saudi premolars on the mesial and distal surface were statistically significant (P < 0.001). CONCLUSIONS: The location of the CDJ varied between the Egyptian and Saudi mandibular first premolars, indicating that this junction is not a fixed point in populations of different countries. PMID- 12887384 TI - An evaluation of the usefulness of two endodontic case assessment forms by general dentists. AB - AIM: To evaluate the use of two forms to assess the risks and difficulty of root canal treatment. METHODOLOGY: Two criterion-based forms, containing 15 and 16 items, respectively, were distributed to 83 general dentists to evaluate the potential difficulty of root-canal treatment. The participants were asked to assess the difficulty of 15 endodontic cases using the Dutch Endodontic Treatment Index (DETI) and the Endodontic Treatment Classification (ETC) forms. A questionnaire was also provided to evaluate the time needed to complete the two forms, their ease of use, the clarity and/or appropriateness of the criteria and any other comments. The outcomes of the assessment were compared with the assessment of each case as carried out by the authors. RESULTS: The response rate was 53%. The DETI was an easy and rapid way to differentiate between uncomplicated and complicated cases. In 13 of the 15 cases, 88-100% of the dentists scored the cases in agreement with the authors. Use of the ETC form was more complicated, as a result of the larger number of variables. However, most respondents recognized the complicated cases, and 91% found the ETC form valuable to help in assessing the difficulty of endodontic cases. CONCLUSIONS: These two forms may help general practitioners to assess the difficulty of endodontic problems and to decide whether to treat the case or to refer it to a specialist. PMID- 12887385 TI - Ultrasound real-time imaging in the differential diagnosis of periapical lesions. AB - AIM: To assess whether the use of real-time ultrasound imaging (echography), together with the application of 'Colour Power Doppler', can help the differential diagnosis of periapical lesions based on their contents. METHODOLOGY: Eleven patients diagnosed with periapical lesions using conventional clinical and radiographic findings and who had been scheduled for endodontic surgery were examined using ultrasound imaging and Colour Power Doppler at the site of the lesions. Lesions were described in each case by an expert echographist, together with an endodontist, according to a scheme described previously. Information on the size of each lesion, its content and its vascular supply was gained and recorded by means of the ultrasound images; then, a tentative differential diagnosis between a cyst and a granuloma was made. The lesions obtained during endodontic surgery were processed and examined with a light microscope. The results from the biopsies were compared with the descriptions made with the ultrasound images. RESULTS: The differential diagnoses between periapical granulomas and cystic lesions, which were based on the echograpic findings, were confirmed by the results of the histopathologic examination in all the 11 cases: four being granulomas and seven being cystic lesions. CONCLUSIONS: Ultrasound real-time imaging is a technique that may help make a differential diagnosis between cysts and granulomas by revealing the nature of the content of a bony lesion. This technique may have further applications in the study of other lesions of the jaws. PMID- 12887386 TI - Osteoblasts and MG-63 osteosarcoma cells behave differently when in contact with ProRoot MTA and White MTA. AB - AIM: To test the hypothesis that MG-63 osteosarcoma cells and primary osteoblasts react differently to ProRoot trade mark MTA (mineral trioxide aggregate) and White MTA by: (i) investigating the attachment of primary osteoblasts and MG-63 osteosarcoma cells to ProRoot trade mark MTA and White MTA; and (ii) comparing the osteogenic behaviour of both cell lines in contact with these endodontic materials. METHODOLOGY: Primary osteoblasts were harvested from foetal rat calvaria by sequential digestion and MG-63 osteosarcoma cells were purchased. Cells were exposed to ProRoot trade mark MTA and White MTA prepared according to the manufacturer's instructions. All samples and controls were prepared in quadruplicate. After 6, 9 and 13 days exposure to MTA, the cells were fixed and prepared for SEM examination. In addition, both the cell types were grown to confluence and exposed to beta-glycerophosphate and dexamethasone to assess mineralized nodule formation as a function of osteogenic behaviour. RESULTS: The number of cells on the surface of the culture dish and on top of the materials increased in all samples throughout the 3 time periods, except for White MTA where no primary osteoblasts were visible on top of the material by the end of 13 days. After exposing cells to differentiation medium nodules were observed in cultures of primary osteoblasts, but not of MG-63 osteosarcoma cells. CONCLUSIONS: Under the conditions of this study, whilst primary osteoblasts initially bound to White MTA, they did not survive on the surface by the end of 13 days. Primary osteoblasts formed mineralized nodules when exposed to differentiation medium, whilst MG-63 cells did not form nodules. As MG-63 cells do not behave osteogenically by forming mineralized nodules, and primary osteoblasts are more sensitive than MG-63 osteosarcoma cells to White MTA in cell culture, primary osteoblasts are more appropriate than MG-63 cells for testing endodontic materials in cell culture. PMID- 12887387 TI - Botulinum toxin type A: myths, facts, and current research. PMID- 12887388 TI - Neurotoxins in the neurobiology of pain. AB - Migraine is a common, chronic, incapacitating, neurovascular disorder that affects an estimated 12% of the population. Understanding the basic mechanisms of pain is important when treating patients with chronic pain disorders. Pain, an unpleasant sensory and emotional experience, is usually triggered by stimulation of peripheral nerves and often associated with actual or potential tissue damage. Peripheral nerve fibers transmit pain signals from the periphery toward the spinal cord or brain stem. The different diameter pain fibers (A and C) vary in the speed of conduction and the type of pain transmitted (eg, sharp versus dull). When stimulated, peripheral pain fibers carrying sensory input from the body enter at different layers of the dorsal horn, which is then propagated toward the thalamus via the spinothalamic tract within the spinal cord. Conversely, sensory input from the face does not enter the spinal cord but enters the brain stem via the trigeminal nerve. This review describes in detail the neurobiological mechanisms and pathways for pain sensation, with a focus on migraine pain. PMID- 12887389 TI - Evidence for antinociceptive activity of botulinum toxin type A in pain management. AB - The neurotoxin, botulinum toxin type A, has been used successfully, in some patients, as an analgesic for myofascial pain syndromes, migraine, and other headache types. The toxin inhibits the release of the neurotransmitter, acetylcholine, at the neuromuscular junction thereby inhibiting striated muscle contractions. In the majority of pain syndromes where botulinum toxin type A is effective, inhibiting muscle spasms is an important component of its activity. Even so, the reduction of pain often occurs before the decrease in muscle contractions suggesting that botulinum toxin type A has a more complex mechanism of action than initially hypothesized. Current data points to an antinociceptive effect of botulinum toxin type A that is separate from its neuromuscular activity. The common biochemical mechanism, however, remains the same between botulinum toxin type A's effect on the motor nerve or the sensory nerve: enzymatic blockade of neurotransmitter release. The antinociceptive effect of the toxin was reported to block substance P release using in vitro culture systems. The current investigation evaluated the in vivo mechanism of action for the antinociceptive action of botulinum toxin type A. In these studies, botulinum toxin type A was found to block the release of glutamate. Furthermore, Fos, a product of the immediate early gene, c-fos, expressed with neuronal stimuli was prevented upon peripheral exposure to the toxin. These findings suggest that botulinum toxin type A blocks peripheral sensitization and, indirectly, reduces central sensitization. The recent hypothesis that migraine involves both peripheral and central sensitization may help explain how botulinum toxin type A inhibits migraine pain by acting on these two pathways. Further research is needed to determine whether the antinociceptive mechanism mediated by botulinum toxin type A affects the neuronal signaling pathways that are activated during migraine. PMID- 12887390 TI - Synaptic transmission: inhibition of neurotransmitter release by botulinum toxins. AB - Botulinum toxin type A, a protein long used in the successful treatment of various dystonias, has a complex mechanism of action that results in muscle relaxation. At the neuromuscular junction, the presynaptic nerve ending is packed with synaptic vesicles filled with acetylcholine, and clustered at the tip of the folds of the postsynaptic muscle membrane are the acetylcholine receptors. Synaptic vesicles fuse with the membrane in response to an elevation of intraneuronal calcium concentration and undergo release of their transmitter by exocytosis. Intracellular proteins that contribute to the fusion of the vesicles with the plasma membrane during exocytosis include synaptosomal protein with a molecular weight of 25 kDa (SNAP-25); vesicle-associated membrane protein (VAMP), also known as synaptobrevin; and syntaxin. Through their proteolytic action on these proteins, botulinum toxins prevent exocytosis, thereby inhibiting the release of acetylcholine. There are 7 serotypes of this toxin-A, B, C1, D, E, F, and G-and each cleaves a different intracellular protein or the same target at distinct bonds. The separate cleavage sites in SNAP-25 for botulinum toxin types A and E contribute to their dissimilar durations of muscle relaxation. This report describes the molecular basis for the inhibition by botulinum toxins of neuroexocytosis and subsequent functional recovery at the neuromuscular junction. PMID- 12887391 TI - Botulinum neurotoxin for the treatment of migraine and other primary headache disorders: from bench to bedside. AB - Botulinum toxin type A, a neurotoxin, is effective for treating a variety of disorders of involuntary muscle contraction including cervical dystonia, blepharospasm, and hemifacial spasm. It inhibits neuromuscular signaling by blocking the release of acetylcholine at the neuromuscular junction. The biological effects of the toxin are transient, with normal neuronal signaling returning within approximately 3 to 6 months postinjection. Recent clinical findings suggest that botulinum toxin type A may inhibit pain associated with migraine and other types of headache. However, the mechanism by which this toxin inhibits pain is not fully understood and is under investigation. Research findings suggest that botulinum toxin type A inhibits the release of neurotransmitters from nociceptive nerve terminals and, in this way, may possess an analgesic effect. A number of retrospective open-label chart reviews and 3 double-blind, placebo-controlled trials have demonstrated that localized injections of botulinum toxin type A significantly reduce the frequency, severity, and disability associated with migraine headaches. Although the majority of patients in these studies experienced no botulinum toxin type A mediated side effects, a small percentage of patients did report transient minor side effects including blepharoptosis, diplopia, and injection-site weakness. Currently, 4 randomized, placebo-controlled, clinical trials are being conducted to evaluate the efficacy, optimal dosing, and side-effect profile of botulinum toxin type A as a novel treatment for migraine and other types of headache. These studies may provide further evidence that botulinum toxin type A is an effective option for the preventive treatment of migraine. PMID- 12887392 TI - Ecology of oral health: a complexity perspective. AB - Health and disease are determined by many interwoven factors. Complexity theory may facilitate an understanding of oral health and disease by studying their complex interplay, and not focusing only on the importance of each individual factor involved. This introduces a theoretical basis for a dynamic, holistic approach to oral health with possible far-reaching consequences for dental education and diagnosis, prevention and treatment of oral diseases. PMID- 12887393 TI - Relationship between dental health and 10-year mortality in a cohort of community dwelling elderly people. AB - Dental examinations were carried out as part of the Evergreen project, which focuses on functional capacity and health among the elderly residents of the city of Jyvaskyla, central Finland. Dental status was examined in 1990 for the whole population born in 1910 (n = 226). Mortality data were collected over 10 yr. The aim of the study was to assess the possible role of dental health as a predictor of mortality. The Kaplan-Meier method was used to analyse survival curves and Cox regression models, with the number of chronic conditions and self-rated health used as covariates in analysing the risks of death. The results showed that the more teeth or filled teeth a subject had, the smaller was their risk for death. The effect of missing teeth was significant after adjusting for the general health variables. Thus, our results support the hypothesis that poor dental health is linked to increased mortality among elderly people. PMID- 12887394 TI - The impact of stomatological disease on oral health-related quality of life. AB - The clinical diagnosis of stomatological disease may indicate its cause and prognosis; however, it says little about the resulting level of impairment from the patient's perspective. The primary objective of this study was to test whether patients attending an outpatient oral medicine clinic would have worse oral health related quality of life (OHR-QoL) compared with the general population. In addition, we aimed to assess whether anxiety or depression could be predicted by OHR-QoL and to explore the relationship between clinical diagnoses, OHR-QoL and anxiety/depression. Data were collected from patients (n = 97) through face-to-face interviews using the Oral Health Impact Profile Short form (OHIP-14) to measure OHR-QoL, the Hospital Anxiety and Depression Scale (HADS) for psychiatric morbidity, and a visual analogue scale for self-rated general health. Age- and sex-matched controls (n = 388) were provided from a normative data set collected in a UK national survey in 1998. Participants had significantly lower OHR-QoL scores than the general population on all domains and overall OHR-QoL scores. Of the variance in anxiety, 55% was predicted by general health ratings and OHR-QoL domains of 'psychological discomfort' and 'psychological disability'. Of the variance in depression, 54% was predicted by general health ratings and OHR-QoL domains of 'functional limitation' and 'social disability'. Patient centred, routine assessment of OHR-QoL provides an additional dimension that may help to improve awareness of the impact of disease on the individual's life and enhance the clinical decision-making process. PMID- 12887395 TI - Burning mouth syndrome: experiences from the perspective of female patients. AB - The aim of this study was to gain a deeper understanding of the development of burning mouth syndrome (BMS). Eighteen females (43-80 yr old), referred to the Clinic of Oral Medicine, formed the study group. Oral examination did not reveal any mucosal lesion or dental pathology, blood analyses of vitamin B12 and serum iron levels were within normal reference values, and no allergies to dental materials were reported. Reported pain/discomfort was 49.4 (mean) on a 100-mm visual analog scale and duration of symptoms was, on average, 8.1 yr (median = 1.5 yr). Taped semistructured interviews were transcribed and analysed in line with grounded theory methodology. In the analysis, a core category emerged, labeled communicating psychological discomfort. This category indicated musings about the meaning of life and interacted with a personality characterized by a strong need for conscientiousness, a life style including long-standing struggle with psychosocial overload, and external social and cultural influences. The debut of the BMS was then preceded by an acutely stressful event. A psychosocial history, in addition to a detailed dental and medical history, seems to be crucial in diagnosing patients. If a psychological disorder and somatization of anxiety and depression is suggested, as indicated in our study, the patient should be offered counseling by a psychologist. PMID- 12887396 TI - The permeability of oral leukoplakia. AB - The significant increase in oral cancer mortality necessitates further research on the mechanisms of tumorigenesis. It was the aim of this study to compare the permeability, lipid composition and histopathological characteristics of oral leukoplakia with non-lesional specimens of the same region in 30 cases as well as 11 specimens originating from healthy control buccal mucosa. The permeability (Kp) of tissue biopsies to tritiated nitrosonornicotine was determined in a continuous through-flow perfusion system, lipids were extracted and identified by thin-layer chromatography, and thickness of epithelium and keratin layer assessed by histopathological methods. Results of the measurements showed that the permeability to the tobacco carcinogen, nitrosonornicotine for leukoplakic tissue was higher than for normal control buccal specimens. Non-lesional areas of buccal mucosa, adjacent to leukoplakias, showed hyperplasia and significantly higher permeability values than both leukoplakic and normal buccal control mucosa. The lipid content of the non-lesional sites was intermediate between the increased values of the leukoplakic lesion and of normal control mucosa. The data strongly suggest that the presence of tobacco in the oral cavity may bring about generalized changes even in regions that do not show leukoplakia. PMID- 12887397 TI - Sugar-starch combinations in food and the relationship to dental caries in low risk adolescents. AB - The aim of this 2-year prospective cohort study was to determine whether food level sugar-starch combinations are predictors of dental caries in a low-risk adolescent population. A total of 645 subjects, aged 12-13 yr at baseline, were recruited from 25 secondary colleges in the north-west region of metropolitan Melbourne, Australia. Examinations to record dental caries status were conducted annually. Dental caries was diagnosed according to the criteria of the World Health Organization. Dietary information was collected by four, continuous 4-d records. Demographic data was collected by parental self-administered questionnaire. A total of 504 subjects provided complete information for analysis. Approximately 37% of subjects experienced an increment in caries. In the multivariate model, only the low sugar-high starch food group was a significant predictor of caries increment on all surfaces and pit and fissure surfaces. For both these surfaces, significant interactions with starch at low sugar and across those clusters with a maximum proportion of sugar and/or starch (that is, high sugar-low starch, medium sugar-medium starch and high sugar-low starch) were found. Sugar-starch interactions may be predictive of caries risk in a low-risk adolescent population. Changing patterns of food consumption and the widespread exposure to various fluoride vehicles are possibly altering the diet dental caries dynamic that once existed. PMID- 12887398 TI - Exclusion of candidate genes in two families with autosomal dominant hypocalcified amelogenesis imperfecta. AB - The amelogenesis imperfectas (AI) are a group of hereditary enamel defects characterized by clinical and genetic diversity. The most common AI types are inherited as autosomal traits. Three mutations of the enamelin (ENAM) gene have been found in cases of autosomal dominant hypoplastic AI. The gene(s) responsible for hypocalcified forms of AI have not been identified, although a number of autosomal genes have been proposed as candidates for AI based on their expression by ameloblasts, including ameloblastin and enamelin (chromosome 4q13.3), tuftelin (chromosome 1q21), enamelysin (chromosome 11q22.3-q23) and kallikrein 4 (chromosome 19q13.3-q13.4). To localize the gene(s) responsible for autosomal dominant hypocalcified AI, we evaluated support for/against linkage of AI to genetic markers spanning five AI candidate genes in two extended families. Our data excluded all proposed candidate gene regions as causal for autosomal dominant hypocalcified AI in these families. These linkage findings provide further evidence for genetic heterogeneity among families with autosomal dominant AI and indicate that, at least, some forms of autosomal dominant hypocalcified AI are not caused by a gene in the five most commonly reported AI candidate genes. PMID- 12887399 TI - Pulp cell responses during hypoxia and reoxygenation in vitro. AB - The purpose of this study was to investigate pulp cell responses during hypoxia and reoxygenation. Pulp tissues obtained from beagle dogs were cultured. In the control group, pulp cells were incubated in normoxic conditions (20% O2) for 1-4 d. In the hypoxia group, pulp cells were incubated under hypoxic conditions (2% O2) for 1-4 d. In the reoxygenation group, pulp cells were first incubated under hypoxic conditions for 24 h, and were then incubated in normoxic conditions (20% O2) for one to three additional days. Cell viability, MTT (3- (4,5 dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) reduction assay, cellular proliferation, and alkaline phosphatase (ALPase) activity were determined. Expression of heat shock protein 70 (HSP70) and vascular endothelial growth factor (VEGF) was analysed by Western blotting. Hypoxia inducible factor-1alpha (HIF-1alpha) in pulp cells was analysed by reverse transcriptase polymerase chain reaction (RT-PCR). The cell growth rate and ALPase activity were significantly higher in the hypoxia group than in the control group. After reoxygenation, cellular proliferation and ALPase activity decreased to the level of the control group while HSP70 expression increased. Hypoxia inducible factor-1alpha expression was detected in pulp cells, and VEGF expression (which is regulated by HIF-1alpha) increased under hypoxic conditions. These results suggest that dynamic responses to hypoxia and reoxygenation occur in pulp cells. PMID- 12887400 TI - Effects of local injection of prilocaine-felypressin on the myocardial oxygen balance in dogs. AB - The authors investigated the effects of felypressin (Fely), a non-adrenergic vasoconstrictor, used together with prilocaine on myocardial oxygen balance. Six open-chest dogs were studied under urethane and alpha-chloralose anesthesia. Systolic arterial pressure, diastolic arterial pressure, mean pulmonary arterial pressure and pulmonary capillary wedge pressure, heart rate, coronary blood flow (CBF), internal and external myocardial oxygen tension (int- or ext-PmO2), and cardiac output were observed. Three doses of Citanest-Octapressin, which contains 3% prilocaine and 0.03 IU ml(-1) Fely (Pri-Fely) - 0.09, 0.18, and 0.3 ml kg(-1)- were injected into the tongue. Observations were performed up to 60 min after the injection. The CBF and int-PmO2 was reduced following the injection of each of the three doses of Pri-Fely. There were negative correlations between the Pri Fely dose per body weight and the maximum reductions in CBF (r = -0.52, P < 0.05), in int-PmO2 (r = -0.78, P < 0.05), and in ext-PmO2 (r = -0.55, P < 0.05), respectively [corrected]. These results suggest that an administration of Fely at doses more than 2.7-5.4 mIU kg(-1) (3-6 cartridges of Pri-Fely) may induce an imbalance between the oxygen supply and demand in myocardial tissues of patients with cardiovascular diseases. PMID- 12887401 TI - Expression and role of RANKL in periodontal ligament cells during physiological root-resorption in human deciduous teeth. AB - Although important roles of receptor activator of NF-kappaB ligand (RANKL) and its receptor (RANK) have been established for osteoclastogenesis and bone resorption, their expression and roles during physiological root resorption remain uncertain. Physiological root resorption for shedding of human deciduous teeth is mediated by osteoclast-like cells (odontoclasts). In this study, we examined the expression of RANKL and osteoprotegerin (OPG), a decoy receptor that prevents RANKL from binding to RANK in human periodontal ligament (PDL) cells during physiological root resorption using immunocytochemistry and reverse transcriptase polymerase chain reaction. The effect of RANKL on root resorbing activity of odontoclasts was evaluated by measuring the size of dissolved area on calcium phosphate-coated coverslips. The PDL cells isolated from either non resorbing deciduous teeth or permanent teeth abundantly expressed OPG, but not RANKL. In contrast, PDL cells derived from resorbing deciduous teeth dominantly expressed RANKL. Human odontoclasts derived from resorbing deciduous teeth expressed both RANKL and RANK. It was observed that RANKL increased odontoclast actin ring formation and resorbing activity in a dose-dependent manner. These results indicate that PDL cells during the root-resorbing state express RANKL but decrease OPG expression. Expression of RANKL likely participates in odontoclastogenesis and activates physiological root resorption. PMID- 12887402 TI - Impulsive compression influences the viscous behavior of porcine temporomandibular joint disc. AB - Traumatic joint injury is known to produce osteoarthritic degeneration in the temporomandibular joint (TMJ). However, little information, is available on its possible effect on the dynamic viscoelastic behavior of the disc. In the present study, impulsive compression was applied to the disc as a model for traumatic joint injury. This was tested in 32 porcine discs. Prior to a dynamic tensile test, impulsive compression with strain rates of 0.01, 0.1 and 1 s(-1) to a final strain of 30% was applied to these discs. Tensile stress was applied in the antero-posterior direction with a wide range of frequencies (0.1-100 Hz). The dynamic E-moduli increased as the loading frequency increased. The dynamic viscosity and elasticity decreased with an increase of strain rate, although the effect on viscosity was greater than on elasticity. The results suggest that strain rate and subsequent peak stress of impulsive compression affect primarily the dynamic viscosity of the disc, and that impulsive compression at high strain rates reduces the resistance of the TMJ disc to stress, resulting in permanent disc deformation and perforation. PMID- 12887403 TI - Surface characteristics of dentin experimentally exposed to hydrofluoric acid. AB - The purpose of this study was to test the effect of hydrofluoric acid (HF) on the surface characteristics of dentin in vitro. Dentin was exposed in 50 human molars and divided into five groups according to different etching schedules: (i) no etching, (ii) 15 s HF, (iii) 15 s H3PO4, (iv) 15 s HF and 15 s H3PO4, (v) 15 s H3PO4 and 15 s HF. Teeth were examined under a scanning electron microscope equipped with energy-dispersive X-ray (EDX) or two layers of fluorescence-labeled primer followed by the composite were applied, and the teeth were sectioned and examined using confocal laser scanning microscopy (CLSM). With scanning electron microscopy, no openings of dentinal tubules were found in groups (i), (ii), and (iv). In group (v) only a few tubules were opened and in group (iii) the smear layer was completely removed and tubules appeared open. The EDX analysis revealed that fluoride was incorporated into the dentin surface when HF was used. With CLSM, distinct hybrid layers could be detected only in group (iii). In group (v) the hybrid layer appeared less established compared with group (iii). No dentin hybridization was found in groups (i), (ii), and (iv). It is concluded that HF has the ability to close the openings of dentin tubules which were opened due to etching by phosphoric acid and cannot dissolve the smear layer. PMID- 12887404 TI - Dissociation between top-down attentional control and the time course of visual attention as measured by attentional dwell time in patients with mild cognitive impairment. AB - Studies of the time course of visual attention have identified a temporary functional blindness to the second of sequentially presented stimuli in that the attentional cost of attending to one visual stimulus may lead to impairments in identifying a second stimulus presented within 500 ms of the first. This phenomenon is known as the attentional blink or attentional dwell time. The neural correlates of the attentional blink and its relationship to mechanisms that control attention are unknown. To examine this relationship we tested healthy controls and subjects in the preclinical stage of Alzheimer's disease, known as mild cognitive impairment (MCI), on a paradigm which affords quantification of both the attentional blink and the top-down control of attention. When subjects were asked to identify both a number and a letter that were rapidly and sequentially presented on a visual display, the detrimental effect that identifying the first stimulus had on the ability to identify the second served as a measure of the attentional blink. When asked to identify only one of the two stimuli, the ability to ignore the first stimulus was a function of their top-down attentional control. The MCI subjects demonstrated a normal attentional dwell time but in contrast they showed impaired top-down attentional control within the same paradigm. This dissociation suggests that these two aspects of visual attention are subserved by different neural systems. The possible neural correlates of these two attentional functions are discussed. PMID- 12887405 TI - Functional characterization of a neuropeptide F-like receptor from Drosophila melanogaster. AB - A cDNA clone encoding a seven-transmembrane domain, G-protein-coupled receptor (NPFR76F, also called GPCR60), has been isolated from Drosophila melanogaster. Deletion mapping showed that the gene encoding this receptor is located on the left arm of the third chromosome at position 76F. Northern blotting and whole mount in situ hybridization have shown that this receptor is expressed in a limited number of neurons in the central and peripheral nervous systems of embryos and adults. Analysis of the deduced amino acid sequence suggests that this receptor is related to vertebrate neuropeptide Y receptors. This Drosophila receptor shows 62-66% similarity and 32-34% identity to type 2 neuropeptide Y receptors cloned from a variety of vertebrate sources. Coexpression in Xenopus oocytes of NPFR76F with the promiscuous G-protein Galpha16 showed that this receptor is activated by the vertebrate neuropeptide Y family to produce inward currents due to the activation of an endogenous oocyte calcium-dependent chloride current. Maximum receptor activation was achieved with short, putative Drosophila neuropeptide F peptides (Drm-sNPF-1, 2 and 2s). Neuropeptide F-like peptides in Drosophila have been implicated in a signalling system that modulates food response and social behaviour. The identification of this neuropeptide F-like receptor and its endogenous ligand by reverse pharmacology will facilitate genetic and behavioural studies of neuropeptide functions in Drosophila. PMID- 12887406 TI - Endogenous rhythm generation in the pre-Botzinger complex and ionic currents: modelling and in vitro studies. AB - The pre-Botzinger complex is a small region in the mammalian brainstem involved in generation of the respiratory rhythm. As shown in vitro, this region, under certain conditions, can generate endogenous rhythmic bursting activity. Our investigation focused on the conditions that may induce this bursting behaviour. A computational model of a population of pacemaker neurons in the pre-Botzinger complex was developed and analysed. Each neuron was modelled in the Hodgkin Huxley style and included persistent sodium and delayed-rectifier potassium currents. We found that the firing behaviour of the model strongly depended on the expression of these currents. Specifically, bursting in the model could be induced by a suppression of delayed-rectifier potassium current (either directly or via an increase in extracellular potassium concentration, [K+]o) or by an augmentation of persistent sodium current. To test our modelling predictions, we recorded endogenous population activity of the pre-Botzinger complex and activity of the hypoglossal (XII) nerve from in vitro transverse brainstem slices (700 micro m) of neonatal rats (P0-P4). Rhythmic activity was absent at 3 mm[K+]o but could be triggered by either the elevation of [K+]o to 5-7 mm or application of potassium current blockers (4-AP, 50-200 micro m, or TEA, 2 or 4 mm), or by blocking aerobic metabolism with NaCN (2 mm). This rhythmic activity could be abolished by the persistent sodium current blocker riluzole (25 or 50 micro m). These findings are discussed in the context of the role of endogenous bursting activity in the respiratory rhythm generation in vivo vs. in vitro and during normal breathing in vivo vs. gasping. PMID- 12887407 TI - Subcellular distribution of L-type Ca2+ channels responsible for plateau potentials in motoneurons from the lumbar spinal cord of the turtle. AB - L-type calcium channels mediate the persistent inward current underlying plateau potentials in spinal motoneurons. Electrophysiological analysis shows that plateau potentials are generated by a persistent inward current mediated by low threshold L-type calcium channels located in the dendrites. As motoneurons express L-type calcium channels of the CaV1.2 and CaV1.3 subtypes, we have investigated the subcellular distribution of these channels using antibody labelling. The plateau generating a persistent inward current is modulated by the activation of metabotropic receptors. For this reason, we also examined the relationship between CaV1.2 and CaV1.3 subunits in motoneurons and presynaptic terminals labelled with antibodies against synapsin 1a. Motoneurons in the spinal cord of the adult turtle were identified as large neurons, immunopositive for choline acetyltransferase, located in the ventral horn. In these neurons, CaV1.2 subunits were present in the cell bodies and axons. Patches of CaV1.3 subunits were seen in association with the cell membrane of the somata and both the proximal and distal dendrites. Double labelling with an antibody against synapsin 1a showed that CaV1.3 subunits, but not CaV1.2 subunits, were always located at synaptic sites. The distribution of CaV1.2 and CaV1.3 strongly suggests that the persistent inward current underlying plateau potentials in spinal motoneurons is mediated by CaV1.3 and not by CaV1.2. Our findings also show that CaV1.3 may be located in the somatic and dendritic membrane adjacent to particular presynaptic terminals. PMID- 12887408 TI - Connexin43 is not expressed in principal cells of mouse cortex and hippocampus. AB - Previous immunofluorescence analyses in mice and rats showed a mainly astrocytic expression of the gap junction protein connexin43 (Cx43) in brain. However, in situ hybridization of murine brain sections suggested strong expression of Cx43 mRNA in hippocampal and cortical pyramidal neurons and Purkinje cells. These findings contrast with recent immunoelectron microscopic studies that excluded prominent Cx43 protein expression in neurons. Both contrasting results could be explained by post-transcriptional control mechanisms. Here we demonstrate by conditional replacement of the Cx43 coding region by a lacZ reporter gene, mimicking transcriptional activity of the Cx43 gene, that Cx43 is not expressed in principal cells of murine brain. This histochemical approach used is not prone to cross-reactivity of mRNA probes or antibodies. Furthermore, we show that in situ hybridization signals, suggested to be specific for Cx43 in mouse neurons, are retained even when the Cx43 coding DNA in neurons is removed by cre-mediated deletion. Our results confirm the previous findings of a mainly astrocytic expression of Cx43 in adult mouse brain and underscore the importance of connexin deficient mice as controls for in situ hybridization studies. We found no evidence for post-transcriptional control of the Cx43 gene in principal neurons. Thus, the synchronized activity of neuronal networks cannot depend on Cx43 containing gap junctions in these cells. PMID- 12887409 TI - Anatomical organization of the telencephalic connections of the parafascicular nucleus in adult and developing rats. AB - The parafascicular nucleus (PFN) of the rat, homologous to the human centre median, is an intralaminar nucleus of the thalamus, classically considered as part of the ascending activating system. We have previously demonstrated that it is also connected to several subcortical nuclei. To obtain a more detailed picture of the connectivity of the PFN, the organization and the topography of the reciprocal parafascicular-telencephalic relationships were studied in both adult and developing rats, using anterograde and retrograde neuronal tracers. In the adult rat, the ascending parafascicular projections were densest to the striatum, dense to the frontal and least dense to cingulate cortex, and were strictly ipsilateral. They displayed a loose topography, with the more medial parafascicular neurons projecting to the medial frontal and cingulate cortex and medial striatum, and the more lateral neurons projecting to the lateral frontal cortex and lateral striatum. All these connections were already present at embryonic day 19. Parafascicular neurons projecting to the telencephalon in adult rats were mostly of the multipolar type, with a few bipolar neurons. In neonatal rats they showed a bipolar morphology at birth; they became mostly multipolar later on, with an increasing complexity of the dendritic arbor up to postnatal day 10. Neurons in the frontal cortex retrogradely labelled from the PFN were more numerous perinatally, and decreased as early as postnatal day 5. The telencephalic connections of the PFN were found to be more discrete and restricted than previously thought, thus suggesting a more specific functional role for the nucleus than cortical recruitment. PMID- 12887410 TI - Binding characteristics of [3H]14-methoxymetopon, a high affinity mu-opioid receptor agonist. AB - The highly potent micro -opioid receptor agonist 14-methoxymetopon (4,5alpha epoxy-3-hydroxy-14beta-methoxy-5beta,17-dimethylmorphinan-6-one) was prepared in tritium labelled form by a catalytic dehalogenation method resulting in a specific radioactivity of 15.9 Ci/mmol. Opioid binding characteristics of [3H]14 methoxymetopon were determined using radioligand binding assay in rat brain membranes. [3H]14-Methoxymetopon specifically labelled a single class of opioid sites with affinity in low subnanomolar range (Ki = 0.43 nm) and maximal number of binding sites of 314 fmol/mg protein. Binding of [3H]14-methoxymetopon was inhibited by ligands selective for the micro -opioid receptor with high potency, while selective kappa-opioids and delta-opioids were weaker inhibitors. 14 Methoxymetopon increased guanosine-5'-O-(3-[35S]thio)-triphosphate ([35S]GTPgammaS) binding with an EC50 of 70.9 nm, thus, providing evidence for the agonist character of this ligand. The increase of [35S]GTPgammaS binding was inhibited by naloxone and selective micro -opioid antagonists, indicating a micro -opioid receptor-mediated action. [3H]14-Methoxymetopon is one of the few nonpeptide mu-opioid receptor agonists available in radiolabelled form up to now. Due to its high affinity and selectivity, high stability and extremely low nonspecific binding (<10%), this radioligand would be an important and useful tool in probing mu-opioid receptor mechanisms, as well as to promote a further understanding of the opioid system at the cellular and molecular level. PMID- 12887411 TI - Enabling role of adenosine A1 receptors in adenosine A2A receptor-mediated striatal expression of c-fos. AB - When striatal neurons are strongly activated they produce adenosine, which activates nearby adenosine A1 receptors (A1Rs) and adenosine A2A receptors (A2ARs). Although the effects of A1R or A2AR activation on neural activity in the striatum have been examined separately, the effects of coactivating both receptors has not been investigated. Using c-Fos immunohistochemistry as an indicator of neural activity, we examined the effects of coactivation of A1Rs and A2ARs on neural activity and their mechanism of interaction in the caudate putamen, nucleus accumbens (NAc) and prefrontal cortex in rats. Administration of a motor-depressant dose of the A2AR agonist CGS 21680 (0.5 mg/kg i.p.) did not significantly induce c-fos expression in any of these brain regions. Administration of a motor-depressant dose of the A1R agonist CPA (0.3 mg/kg, i.p.) produced a small but significant induction of c-fos expression only in the shell of the NAc. Coadministration of CGS 21680 and CPA produced a synergistic induction of c-fos expression in the caudate-putamen, cingulate cortex, and especially the NAc. In the shell of the NAc administration of CPA significantly decreased extracellular dopamine levels measured by in vivo microdialysis and blocked CGS 21680-induced increases in dopamine levels. Because it has been previously shown that activation of dopamine D2 receptors (D2Rs) by endogenous dopamine blocks A2AR-mediated c-fos expression, it is hypothesized that the enabling role of A1Rs in A2AR-mediated striatal c-fos expression is related to the A1R-mediated inhibition of dopamine release. PMID- 12887412 TI - Dopamine and adenosine mediate substance P-induced depression of evoked IPSCs in the rat nucleus accumbens in vitro. AB - The major projection cells of the nucleus accumbens (NAc) are under a strong inhibitory influence from GABAergic afferents and depend on afferent excitation to produce their output. We have earlier reported that substance P (SP), a peptide which is colocalized with GABA in these neurons, depresses excitatory synaptic transmission in this nucleus (Kombian, S.B., Ananthalakshmi, K.V.V., Parvathy, S.S. & Matowe, W.C. (2003) J. Neurophysiol., 89, 728-738). In order to better understand the role of this peptide in the synaptic physiology of the NAc, it is important to determine its effects on inhibitory synaptic responses. Using whole-cell recording in rat forebrain slices, we show here that SP also depresses evoked inhibitory postsynaptic currents (IPSCs) in the NAc via intermediate neuromodulators. SP caused a partially reversible, dose-dependent decrease in evoked IPSC amplitude. This effect was present without measurable changes in the holding current, input resistance of recorded cells or decay rate (tau) of IPSCs. It was mimicked by a neurokinin-1 (NK1) receptor-selective agonist, [Sar9, Met (O2)11]-SP, and blocked by an NK1 receptor-selective antagonist, L 732 138. The SP-induced IPSC depression was prevented by SCH23390, a dopamine D1-like receptor antagonist and by 8-cyclopentyltheophylline, an adenosine A1 receptor blocker. Furthermore, the SP effect was also markedly attenuated by exogenous adenosine, dipyridamole, rolipram and barium. These data show that SP, acting on NK1 receptors, depresses inhibitory synaptic transmission indirectly by enhancing extracellular dopamine and adenosine levels. SP therefore acts in the NAc to modulate both excitatory and inhibitory afferent inputs using the same mechanism(s). PMID- 12887413 TI - Distribution of central sensory axons in transgenic mice overexpressing nerve growth factor and lacking functional p75 neurotrophin receptor expression. AB - This study examined the roles of nerve growth factor (NGF) and the p75 neurotrophin receptor (p75NTR) in the growth of dorsal root ganglion (DRG) central processes in the dorsal horn. Two genetically modified mouse strains were used: transgenic mice that overexpress NGF in the CNS under the control of the glial fibrillary acidic protein promoter, and p75NTR exon III null mutant mice that express a hypomorphic form of this receptor. In both NGF transgenic and nontransgenic mice with hypomorphic expression of p75NTR, there is a significant loss of DRG neurons compared to mice with normal p75NTR expression. This reduction in neuron number has been shown to underlie a corresponding decrease in peripheral nociceptive sensory innervation. Within the CNS, however, nociceptive innervation of the dorsal horn appears to be unaffected by hypomorphic expression of p75NTR. Comparisons of calcitonin gene-related peptide immunoreactivity in the dorsal horn revealed that the area occupied by DRG central processes was not significantly different between p75NTR hypomorphic mice and wild-type siblings, or between NGF transgenic mice with either hypomorphic or normal expression of p75NTR. We propose that DRG central processes arborize extensively in both NGF transgenic and nontransgenic p75NTR hypomorphic mice in order to compensate for the loss of DRG neurons and restore dorsal horn innervation to normal levels. We also present evidence suggesting that NGF plays only a minor role in the growth of DRG central processes. PMID- 12887414 TI - Substrates and routes of migration of early generated neurons in the developing rat thalamus. AB - We investigated the substrates supporting neuronal migration, and its routes, during early thalamic development in the rat. Neurons and axonal and glial fibres were identified in embryos with single and double immunohistochemistry; dynamic data were obtained with cell tracers in short-term organotypic cultured slices. The earliest thalamic neurons, originating from the ventricular neuroepithelium between embryonic days 13 and 15, include those of the reticular thalamic nucleus. At this developmental stage, calretinin, calbindin or gamma-aminobutyric acid immunostaining revealed both radially and nonradially orientated neurons in the region of reticular thalamic migration, between the dorsal and ventral thalamic primordia. In cultured slices, injections of fluorescent dyes in the neuroepithelium labelled neurons in a migratory stream along radial glia in the same zone. Some labelled fusiform cells departed from this radial trajectory along orthogonal routes within the dorsal thalamus. Confocal microscopy revealed nonradially orientated neurons in close apposition with a fibre system parallel to the lateral thalamic surface. These fibres expressed axonal markers, including the intermediate filament protein alpha-internexin and a polysialylated form of neuronal cell adhesion molecule. Active migration of nonradially orientated neurons along neuronal substrates was confirmed in living cultured slices. In addition, in vitro and ex vivo experiments revealed neurons migrating tangentially in association with glial fibres. These results provide novel evidence that: (i) early generated thalamic neurons follow nonradial routes in addition to glia-linked radial migration; and (ii), nonradially migrating thalamic neurons move along both glial and axonal substrates, which could represent a distinctive feature of thalamic development. PMID- 12887415 TI - Xenopus embryonic spinal neurons recorded in situ with patch-clamp electrodes- conditional oscillators after all? AB - The central pattern generator for swimming Xenopus embryo is organized as two half-centres linked by reciprocal inhibition. Microelectrode recordings suggest that Xenopus neurons are poorly excitable, necessitating a key role for postinhibitory rebound in the operation of the central pattern generator. However the Xenopus central pattern generator seems unusual in that the component neurons apparently have no intrinsic or conditional rhythmogenic properties. We have re examined the firing properties of Xenopus embryo spinal neurons by making patch clamp recordings in situ from intact spinal cord. Recordings made from 99 neurons were divided into three groups. Central pattern generator neurons overwhelmingly (44/51) fired trains of action potentials in response to current injection. Just over half of the sensory interneurons (13/22) also fired trains of action potentials. Neurons that received no synaptic inputs during swimming mostly fired just one or two action potentials (22/26). Thirty-four neurons were identified morphologically. Commissural (8/12) and descending (6/6) interneurons, key components of the spinal central pattern generator, fired repetitive trains of action potentials during current injection. Neurons that were not part of the central pattern generator did not demonstrate this preponderance for repetitive firing. Analysis of the interspike intervals during current injection revealed that the majority of central pattern generators, descending and commissural interneurons, could readily fire at frequencies up to twice that of swimming. We suggest that Xenopus neurons can be considered as conditional oscillators: in the presence of unpatterned excitation they exhibit an ability to fire rhythmically. This property makes the Xenopus embryonic central pattern generator more similar to other model central pattern generators than has hitherto been appreciated. PMID- 12887416 TI - Cell type-dependent expression of HCN1 in the main olfactory bulb. AB - In many brain regions, hyperpolarization-activated cationic currents (Ih) are involved in the generation of rhythmic activities, but the role of Ih in olfactory oscillations remains unclear. Knowledge of the cellular and subcellular distributions of hyperpolarization-activated and cyclic nucleotide-gated channel (HCN) subunits is necessary for understanding the role of Ih in olfactory network activities. Using light microscopic immunocytochemistry, we demonstrate strong HCN1 labelling of the glomerular layer and moderate staining of granule cell, internal and external plexiform layers of the rat main olfactory bulb. In the glomerular layer, among many unlabelled neurons, two distinct subpopulations of juxtaglomerular cells are labelled. Approximately 10% of the juxtaglomerular cells strongly express HCN1. These small diameter cells are immunoreactive for GABA and comprise a subpopulation of periglomerular cells. An additional subset of juxtaglomerular cells ( approximately 1%) expresses low levels of HCN1. They are large in diameter, GABA immunonegative but immunopositive for vesicular glutamate transporter 2, characterizing them as external tufted cells. Quantitative immunogold localization revealed that the somatic plasma membranes of periglomerular cells contain approximately four times more HCN1 labelling than those of external tufted cells. Unlike in cortical pyramidal cells, immunogold density for HCN1 does not significantly differ in somatic and dendritic plasma membranes of external tufted cells, indicating that post-synaptic potentials arriving at proximal and distal dendrites are modulated by the same density of Ih. Our results demonstrate a cell type-dependent expression of HCN1 in the olfactory bulb and predict a differential contribution of distinct juxtaglomerular cell types to network oscillations. PMID- 12887417 TI - Insulin-like growth factor II mRNA is expressed in neurones of the brain of the bony fish Oreochromis mossambicus, the tilapia. AB - The physiological meaning of insulin-like growth factor II (IGF-II) is still enigmatic. IGF-II occurs in the adult mammalian brain where it is expressed in the mesodermal portion of the choroid plexus and the meninges, but results on its presence in cells of neuroepithelial origin are controversial. However, IGF-II mRNA is transiently expressed in neurones during mammalian early development. In bony fish, IGF-II mRNA is also present in the adult brain but nothing is known about its synthesis sites. Thus, the present study using in situ hybridization with digoxigenin-labelled RNA species-specific probes investigates the cellular distribution of IGF-II mRNA in the adult brain of a bony fish, the tilapia (Oreochromis mossambicus). As in mammals, IGF-II mRNA was strongly expressed in the choroid plexus and meninges. Thus, IGF-II synthesis by choroid plexus and meninges seems to have a long evolutionary history and may be common to all vertebrates. However, as shown by the detailed investigation of landmark nuclei and regions, IGF-II mRNA occurred also in numerous neurones at all levels of the tilapia brain. The distinct localization of IGF-II mRNA in neurones might indicate that neuronal IGF-II acts as transmitter or modulator. However, the widespread occurrence of the IGF-II-producing neurones argues against this assumption and most probably suggests that IGF-II plays a role in the differentiation, maintenance and regeneration of neurones. It is further assumed that the sustained neuronal IGF-II expression in the brain of the adult tilapia correlates with continued post-embryonic up to life-long brain growth as has been shown in many teleost fishes. PMID- 12887418 TI - Identification of circadian brain photoreceptors mediating photic entrainment of behavioural rhythms in lizards. AB - We have shown previously that in ruin lizards (Podarcis sicula) the ablation of all known photoreceptive structures (lateral eyes, pineal and parietal eye) in the same individual animal does not prevent entrainment of their circadian locomotor rhythms to light. The present study was aimed at identifying the circadian brain photoreceptors mediating entrainment. For this purpose, we looked for opsin expression in the brain by means of immunocytochemistry. Using anti cone-opsin antiserum CERN 874 we have localized photoreceptors in the periventricular area of hypothalamus, near the third cerebral ventricle. We also cloned a brain opsin cDNA that, on the basis of the deduced amino acid sequence, appears to belong to the RH2 class of cone-opsins. We named the cloned cone-opsin Ps-RH2. To examine whether brain cone-opsins mediate photic entrainment of circadian locomotor rhythms, we performed post-transcriptional inactivation experiments by injecting an expression eukaryotic vector transcribing the antisense cone-opsin Ps-RH2 mRNA in the third cerebral ventricle of pinealectomized-retinectomized lizards previously entrained to a light-dark (LD) cycle. Injections of the antisense construct abolished photic entrainment of circadian locomotor rhythms of pinealectomized-retinectomized lizards to the LD cycle for 6-9 days. CERN 874 completely failed to label cells within the periventricular area of hypothalamus of brains injected with antisense construct. Thus, abolishment of photic entrainment is due to inactivation of endogenous brain cone-opsins mRNA. The present results demonstrate for the first time in a vertebrate that brain cone-opsins are part of a true circadian brain photoreceptor participating in photic entrainment of behavioural rhythms. PMID- 12887419 TI - The effect of urocortin on ingestive behaviours and brain Fos immunoreactivity in mice. AB - The influence of urocortin (UCN) on ingestive behaviours and brain neural activity, as measured immunohistochemically by the presence of Fos protein, was determined in mice. Rat UCN was administered by continuous intracerebroventricular (ICV) or subcutaneous (SC) infusion. ICV infusion of UCN (100 ng/h, 14 days) transiently reduced daily food and water intakes (days 1-4) but body weight was reduced from day 2 into the post-infusion period. Sodium intake was reduced from day 3 to the end of infusion. SC infusion of UCN caused similar but smaller reductions in food and water intakes and body weight, without change in sodium intake. In separate experiments, Fos immunoreactivity was increased in several brain nuclei known to be involved in the control of body fluid and energy homeostasis, e.g. central nucleus of the amygdala, median preoptic nucleus, bed nucleus of the stria terminalis and arcuate nucleus. Increased Fos expression was similar for ICV and SC infusions when measured on days 2-3 or 6-7 of infusion. In conclusion, increases of brain activity by UCN may be associated with stimulation of adrenocorticotrophic hormone release and sympathetic nervous activity, but increases may also indicate suppression of ingestive behaviours by stimulating central inhibitory mechanisms located in areas known to control body fluid and energy homeostasis. PMID- 12887420 TI - Enkephalin contributes to the locomotor stimulating effects of 3,4-methylenedioxy N-methylamphetamine. AB - 3,4-methylenedioxy-N-methylamphetamine (MDMA, 'Ecstasy') is a potent inhibitor of serotonin uptake, which induces both an increase in locomotion and a decrease in exploratory activity in rodents. Serotonin 5-HT1B receptors, located on the terminals of striatal efferent neurons, have been suggested to mediate these motor effects of MDMA. Striatal neurons projecting to the globus pallidus contain met-enkephalin, whilst those projecting to the substantia nigra contain substance P. We therefore analysed the levels of both peptides using radioimmunocytochemistry after MDMA administration (10 mg/kg, 3 h) in wild-type and 5-HT1B receptor knockout mice. Our results demonstrate that MDMA induces a decrease in pallidal met-enkephalin immunolabelling in wild-type, but not in 5 HT1B receptor knockout mice. Similar results were obtained following treatment with the 5-HT1A/1B agonist RU24969 (5 mg/kg, 3 h), suggesting that activation of 5-HT1B receptors leads to a reduction in met-enkephalin levels in the globus pallidus. In contrast, MDMA had no effect on the nigral substance P levels. We have previously shown that both MDMA and RU24969 fail to stimulate locomotor activity in 5-HT1B receptor knockout mice. Our present data indicate that the opioid antagonist naloxone suppressed the locomotor effects of MDMA. This study is the first to demonstrate that Enk contributes to MDMA-induced increases in locomotor activity. Such an effect may be related to the 5-HT control of pallidal met-enkephalin levels via the 5-HT1B receptors. PMID- 12887421 TI - Gene expression in dopamine and GABA systems in an animal model of schizophrenia: effects of antipsychotic drugs. AB - We used in situ hybridization histochemistry to assess expression of dopamine receptors (D1R, D2R and D3R), neurotensin, proenkephalin and glutamate decarboxylase-67 (GAD67) in the prefrontal cortex, striatum, and/or nucleus accumbens in adult rats with neonatal ventral hippocampal (VH) lesions and in control animals after acute and chronic treatment with antipsychotic drugs clozapine and haloperidol. We also acquired these measures in a separate cohort of treatment-naive sham and neonatally VH-lesioned rats used as an animal model of schizophrenia. Our results indicate that the neonatal VH lesion did not alter expression of D1R, D3R, neurotensin or proenkephalin expression in any brain region examined. However, D2R mRNA expression was down-regulated in the striatum, GAD67 mRNA was down-regulated in the prefrontal cortex and prodynorphin mRNA was up-regulated in the striatum of the VH-lesioned rats as compared with sham controls. Antipsychotic drugs did not alter expression of D1R, D2R or D3R receptor mRNAs but elevated neurotensin and proenkephalin expression in both groups of rats; patterns of changes were dependent on the duration of treatment and brain area examined. GAD67 mRNA was up-regulated by chronic antispychotics in the nucleus accumbens and the striatum and by chronic haloperidol in the prefrontal cortex in both sham and lesioned rats. These results indicate that the developmental VH lesion changed the striatal expression of D2R and prodynorphin and robustly compromised prefrontal GAD67 expression but did not modify drug induced expression of any genes examined in this study. PMID- 12887422 TI - Viral vector-mediated gene transfer of the vole V1a vasopressin receptor in the rat septum: improved social discrimination and active social behaviour. AB - This study explores the effects of enhancing vasopressin V1a receptor expression in the septum using viral vector-mediated gene transfer on social discrimination and social interactions. Bilateral infusion of an adeno-associated viral vector containing the prairie vole V1a receptor gene (V1aR-AAV) regulated by a neuron specific enolase promoter resulted in a stable increase in V1a receptor binding density in the rat septum without affecting oxytocin receptor density. Control animals were infused with a vector expressing the lacZ gene. In a social discrimination paradigm, only V1aR-AAV-treated animals succeeded in discriminating a previously encountered from a novel juvenile after an interexposure interval (IEI) of more than 2 h, demonstrating the functional incorporation of the vole V1a receptor in the rat septal circuits underlying short-term memory processes. Microdialysis administration of synthetic vasopressin during the first juvenile exposure, used to mimic intraseptal release patterns of the neuropeptide, produced similar prolongations in recognition (up to an IEI of 24 h) in both V1aR-AAV and control animals. Septal microdialysis administration of a selective V1a, but not oxytocin, receptor antagonist in both groups prevented discrimination even after an IEI of as short as 0.5 h, confirming the specificity of the vole V1a receptor involvement in social discrimination abilities. In addition, active social interactions were found to be increased among V1aR-AAV rats compared to controls. Viral vector-mediated gene transfer provides a valuable tool for studies on the role of localized gene expression on behavioural parameters. PMID- 12887423 TI - Unilateral dorsal column and rubrospinal tract injuries affect overground locomotion in the unrestrained rat. AB - The purpose of this study was to determine the importance of the rubrospinal pathway and the ascending components of the dorsal column for overground locomotion in adult, unrestrained rats. The dorsal column (excluding the corticospinal tract), the rubrospinal tract or both were damaged unilaterally in rats at the level of the upper cervical spinal cord. Behavioural analysis consisted of skilled locomotion (an evaluation of footslips during ladder walking), a paw usage task and the assessment of ground reaction forces during unrestrained locomotion. All lesioned animals used the forepaw ipsilateral to the lesions less while rearing. Animals with dorsal column injuries used the forelimb contralateral to the spinal injury significantly more while rearing compared with uninjured animals. All lesioned animals produced more footfalls while crossing the ladder compared with uninjured animals. All injuries, regardless of the pathway affected, resulted in significant alterations in body weight support and reduced braking forces from the forelimb ipsilateral to the injury during overground locomotion. Animals typically bore less weight on the hindlimb ipsilateral to the lesion compared with the hindlimb contralateral to the spinal injury. Taken together with previously published work, our data indicate that the rubrospinal and dorsal column pathways are important for forelimb support while rearing and for skilled locomotion. Additionally, the ascending dorsal column pathways and the rubrospinal tract play a role during flat surface overground locomotion and combined damage to these pathways does not alter the acquired gait. PMID- 12887424 TI - Improved reversal learning and altered fear conditioning in transgenic mice with regionally restricted p25 expression. AB - Cleavage of the cyclin-dependent kinase 5 activator p35 generates the protein fragment p25, which accumulates in the forebrain of patients with Alzheimer's disease. Although p25 expression has been suggested to affect learning and memory, this hypothesis has not been tested to date. To investigate the role of p25 in hippocampus-dependent learning and memory we have generated transgenic mice expressing p25 preferentially in postnatal forebrain. p25 expression was highest in hippocampus where it averaged approximately 33% of endogenous p35 expression. This low level of p25 expression did not seem to result in hyperphosphorylation of tau, but increased the phosphorylation of neurofilament M and enhanced the expression of tau protein. These molecular changes did not correlate with neurodegeneration or motor abnormalities. In the Morris water maze the p25 mutants were normal in learning an initial platform location, but surprisingly reversal learning was improved when the platform position was changed. The p25 mutants were normal in contextual fear conditioning. However, when trained with a tone presentation the mutants showed reduced contextual conditioning and enhanced tone fear conditioning. We conclude that low p25 expression has pleiotropic effects on learning and memory. As p25 expression can improve learning and memory, p25 formation could be a compensatory mechanism for learning and memory deficits in Alzheimer's disease. PMID- 12887425 TI - Tonotopic representation of missing fundamental complex sounds in the human auditory cortex. AB - The N1m component of the auditory evoked magnetic field in response to tones and complex sounds was examined in order to clarify whether the tonotopic representation in the human secondary auditory cortex is based on perceived pitch or the physical frequency spectrum of the sound. The investigated stimulus parameters were the fundamental frequencies (F0 = 250, 500 and 1000 Hz), the spectral composition of the higher harmonics of the missing fundamental sounds (2nd to 5th, 6th to 9th and 10th to 13th harmonic) and the frequencies of pure tones corresponding to F0 and to the lowest component of each complex sound. Tonotopic gradients showed that high frequencies were more medially located than low frequencies for the pure tones and for the centre frequency of the complex tones. Furthermore, in the superior-inferior direction, the tonotopic gradients were different between pure tones and complex sounds. The results were interpreted as reflecting different processing in the auditory cortex for pure tones and complex sounds. This hypothesis was supported by the result of evoked responses to complex sounds having longer latencies. A more pronounced tonotopic representation in the right hemisphere gave evidence for right hemispheric dominance in spectral processing. PMID- 12887426 TI - Repeated withdrawal from ethanol impairs acquisition but not expression of conditioned fear. AB - Repeated withdrawal from ethanol impairs acquisition of conditioned fear [Stephens, D.N., Brown, G., Duka, T. & Ripley, T.L. (2001) Eur. J. Neurosci., 14, 2023-2031]. This study further examined the effect of repeated withdrawal from ethanol on the expression and acquisition of fear conditioning. Following training, presentation of a cue associated with footshock (CS+) resulted in a suppression of operant responding for food reinforcement. In different groups, shock thresholds were manipulated to give weak or severe behavioural suppression. Rats were subsequently chronically treated with ethanol-containing liquid diet either continuously (single withdrawal) or with three withdrawal periods (repeated withdrawal). Ethanol treatment and withdrawal had no effect on conditioned suppression of responding tested 2 weeks after the final withdrawal, at either shock intensity. Nevertheless, extinction of conditioned fear was impaired in the repeated withdrawal group exposed to the higher shock intensity. In the high intensity group, the stimulus-shock association was then reversed, so that the previously neutral conditioned stimulus (CS-) became the CS+. Acquisition of suppression to the new CS+ was significantly less in the animals previously given repeated experience of withdrawal, confirming our previous finding. Thus, repeated withdrawal from ethanol lead to disruption in the acquisition of fear conditioning but had no effect on retrieval of an association formed prior to the ethanol-withdrawal experiences. PMID- 12887427 TI - Visual receptive field properties of neurons in the caudate nucleus. AB - Visual single-unit activity was recorded in the caudate nucleus of halothane anaesthetized, immobilized, artificially respirated cats. Visually sensitive neurons were found in the dorsolateral part of the caudate body. A majority of the units responded optimally to small spot-like stimuli moving with velocities between 30 and 120 degrees /s. The receptive field of these units is large: it covers a major part of both the contra- and ipsilateral visual hemifields. No signs of retinotopy were observed. Most of the neurons display directional selectivity and are narrowly tuned to the direction of the moving stimulus. These physiological properties are consistent with recent morphological results that reveal multiple connections of the caudate nucleus with the superior colliculus through tecto-extrageniculo-thalamic pathways in the mammalian brain. PMID- 12887428 TI - Task-dependent intermanual coupling of 8-Hz discontinuities during slow finger movements. AB - During slow finger movements, small discontinuities are visible at approximately 8-10 Hz. We have recorded from eight normal subjects whilst they performed index finger flexion-extension movements with the left and right hand. Movements were either performed in-phase (both fingers flexing or extending together), or anti phase (flexion on one side coinciding with extension on the other). Coherence calculated between left and right finger velocity was significantly above zero at approximately 8 Hz for the in-phase condition, but was significantly smaller for anti-phase movements (mean coherence across all subjects at 8.7 Hz was 0.031 for in-phase, 0.010 for anti-phase). We also calculated a 'phase coherence' measure which, unlike conventional coherence analysis, was sensitive only to phase synchronization and not to amplitude co-variations. For the in-phase task, phase coherence values were smaller than coherence, but still significantly different from zero around 8 Hz; for the anti-phase task, phase coherence was not significant in this band. Measures calculated from EMG recordings yielded similar conclusions to those using finger velocity, indicating that the results were not simply due to mechanical cross-talk. Neural oscillators generating approximately 8-Hz movement discontinuities on each side of the body are therefore selectively coupled during the in-phase task. A wavelet-based analysis further suggested that intermanual coupling modulated during in-phase task performance; coupling was maximal at the start and the end of a movement. We conclude that the systems producing approximately 8-Hz movement discontinuities and those responsible for intermanual coupling are likely to share common neural elements. PMID- 12887429 TI - Treatment of infantile spasms: the ideal and the mundane. PMID- 12887430 TI - Protection provided by cyclosporin A against excitotoxic neuronal death is genotype dependent. AB - PURPOSE: Previous studies have shown that the immunosuppressant cyclosporin A (CsA), a specific blocker of the mitochondrial permeability transition (MPT) pore, can dramatically ameliorate the selective neuronal necrosis resulting from ischemia-reperfusion, traumatic brain injury, and N-methyl-d-aspartate (NMDA) evoked neurotoxicity. The purpose of this study was to determine whether two different immunosuppressants, CsA and FK-506, could ameliorate the neuronal damage observed after kainate-induced seizures in strains that are differentially susceptible to excitotoxin-induced cell death. METHODS: Excitotoxin-resistant (C57BL/6) or -susceptible (FVB/N) mice were administered kainate alone (30 mg/kg), CsA alone (5, 10, or 20 mg/kg), or one of the immunosuppressants (CsA, 5 mg/kg or 10 mg/kg; FK-506, 0.5 mg/kg) followed by kainate. After drug administration, mice were monitored continuously for the onset and extent of seizure activity. After a survival of 7 days, animals were assessed for hippocampal damage. RESULTS: Whereas CsA alone induced no epileptogenic effects and both immunosuppressants were without effect on the induction of kainate induced seizures, administration of CsA to excitotoxin-susceptible mice (FVB/N) virtually eliminated neuronal cell death. In contrast, induction of neuronal cell death was evident when CsA was administered to excitotoxin-resistant mice (C57BL/6). Administration of FK-506, another commonly used immunosuppressant, which lacks an effect on the MPT, had no effect on modification of susceptibility to kainate-induced cell death in either strain. CONCLUSIONS: As our data show differential protection of hippocampal neurons against excitotoxic cell death by pretreatment with CsA, these results suggest that strain-dependent differences in mitochondrial integrity and function may exist. PMID- 12887431 TI - Interactions of lamotrigine with topiramate and first-generation antiepileptic drugs in the maximal electroshock test in mice: an isobolographic analysis. AB - PURPOSE: The study investigated the types of interactions between lamotrigine (LTG) and first-generation antiepileptic drugs (AEDs) or topiramate (TPM) with isobolographic analysis. METHODS: Anticonvulsant and adverse-effect profiles of combinations of LTG with other AEDs, at fixed ratios of 1:3, 1:1, and 3:1, were evaluated in the maximal electroshock (MES)-induced seizures and the chimney test (motor performance) in mice, which allowed the determination of benefit indices (BIs) for individual combinations. RESULTS: Combinations of LTG with TPM or valproate (VPA), at fixed ratios of 1:1, were significantly supraadditive (synergistic) in the MES test and, simultaneously, subadditive (antagonistic) in the chimney test, showing the best profile for AED combinations. In contrast, combinations between LTG and carbamazepine (CBZ), in terms of antiseizure protection against MES, were subadditive (antagonistic) and additive in the chimney test, resulting in unfavorable AED combinations. Moreover, the combination of LTG with phenobarbital (PB), at a fixed ratio of 1:1, despite synergy in the MES test, also was synergistic in the chimney test, resulting in a modest BI for AED combination. LTG combined with phenytoin was additive in both the MES and chimney tests in mice. The remaining combinations, at fixed ratios not mentioned earlier, also showed an average BI for AED combinations. Furthermore, LTG combined with all studied AEDs did not affect long-term memory in mice. None of the AEDs influenced the free plasma level of LTG, whereas LTG slightly reduced the free plasma concentration of PB. CONCLUSIONS: Interactions between LTG and TPM or LTG and VPA at a fixed ratio of 1:1 might be profitable from a preclinical point of view, displaying the most optimal BI. PMID- 12887432 TI - Valproate inhibits the conversion of testosterone to estradiol and acts as an apoptotic agent in growing porcine ovarian follicular cells. AB - PURPOSE: Long-term valproate (VPA) treatment has been associated with hyperandrogenism and polycystic ovaries in women with epilepsy. The exact mechanisms of action of the drug on sex steroid hormone function are still unsettled. The aim of the present study was to investigate the action of VPA on basal and gonadotropin-stimulated steroid secretion in porcine ovarian follicular cells and to measure the conversion of testosterone to estradiol. Second, the action of VPA on proliferation and apoptosis of follicular cells was investigated. METHODS: Small and medium follicles were obtained from pig ovaries on days 8-10 and 14-16 of the estrus cycle. Both follicular compartments, theca and granulosa cells, were cultured as a coculture resembling follicles in vivo. VPA in concentrations of 100 and 250 micrg/ml was added to the control or gonadotropin-stimulated cultures. RESULTS: VPA caused a significant increase in basal and luteinizing hormone (LH)-stimulated testosterone secretion from small follicles, whereas in medium follicles, an increased basal but decreased LH stimulated testosterone secretion was found. VPA caused decreased basal and follicle-stimulating hormone (FSH)-stimulated estradiol secretion by small follicles, whereas only the higher concentration decreased estradiol secretion in medium follicles. The conversion of testosterone to estradiol by small follicles was decreased under the influence of VPA in testosterone-alone and in testosterone-plus-FSH-stimulated cultures, whereas this was seen at only the higher VPA concentration in medium follicles. VPA had no effect on cell proliferation and viability, whereas in a dose-dependent manner, VPA increased caspase-3 activity. CONCLUSIONS: VPA affected steroidogenesis in both unstimulated and gonadotropin-stimulated porcine ovarian follicular cells and inhibited the conversion of testosterone to estradiol. In addition, VPA may act as an apoptotic agent in both small and medium-sized follicles. PMID- 12887433 TI - Altered hippocampal expression of neuropeptides in seizure-prone GALR1 knockout mice. AB - PURPOSE: Mice carrying a deletion of the GALR1 galanin receptor have recently showed spontaneous seizure phenotype with 25% penetrance. To better understand the role of neuropeptides, which are known to undergo complex plasticity changes with development of epileptic seizures, we characterized their expression in the hippocampal formation in GALR1- knockout (-KO) mice with or without seizures and in wild-type (WT) mice. METHODS: Immunohistochemistry and in situ hybridization were used to study expression of galanin, neuropeptide Y (NPY), substance P, enkephalin, dynorphin, and cholecystokinin (CCK). RESULTS: In GALR1-KO mice that had been displaying seizures, a strong upregulation of galanin immunoreactivity (ir) and messenger RNA (mRNA) was found in the polymorph layer of the dentate gyrus; galanin-ir also appeared in a dense fiber network in the supragranular layer. A strong upregulation of enkephalin was found in the granule cells/mossy fibers, whereas dynorphin mRNA levels were modestly decreased. NPY was strongly expressed in the granule cells/mossy fibers, and an increase of NPY mRNA levels in the polymorph cells was paralleled by an increase of NPY-ir in the molecular layer. An upregulation of substance P-ir was confined to the fibers in the granule and molecular layers, whereas substance P mRNA was increased in the cells of the polymorph layer. Both CCK-ir and mRNA were strongly downregulated in the granule cell/mossy fiber system, but CCK-ir appeared increased in the supragranular and molecular layers. No changes in neuropeptide-ir were found in GALR1-KO mice not displaying seizures. CONCLUSIONS: Complex changes in neuropeptide expression in some principal hippocampal neurons and interneurons appear as a characteristic feature of the spontaneous-seizure phenotype in GALR1 KO mice. However, to what extent causal relations exist between this "epilepsia peptidergic profile" and development of seizures requires further clarification. PMID- 12887435 TI - Assessment of CNS effects of antiepileptic drugs by using quantitative EEG measures. AB - PURPOSE: Antiepileptic drugs (AEDs) can be associated with adverse neurologic effects including cognitive dysfunction. Objective methods for recognizing AED effects on the brain could be valuable for long-term management. We compared quantitative EEG measures and cognitive tests in a group of patients beginning or ending AED therapy. METHODS: Subjects included 20 patients beginning AED therapy (AEDon), 12 patients stopping AED therapy (AEDoff), 33 patient controls receiving stable AED therapy (AEDco), and 73 healthy controls (Nco). All subjects underwent structured EEG recording and a cognitive test battery before change in AED dose and again 12-16 weeks later, >or=4 weeks after the last dose change. Four occipital EEG measures (peak frequency, median frequency, relative theta and delta power) were analyzed. Cognitive test changes were scored by using test retest regression equations based on the Nco subjects. Wilcoxon tests were used for two-group comparisons. RESULTS: AEDons had a significant decrease, and AEDoffs, a significant increase in the peak frequency of the EEG rhythm, as compared with controls. Results for median frequency and theta power were similar. Change in the EEG peak frequency correlated with an aggregate cognitive change measure (r2= 0.71; p < 0.001), individual cognitive measures, and subjective complaints. Of the combined AEDon/AEDoff patients, 58% exceeded the 95% confidence interval for test-retest change in EEG peak frequency. CONCLUSIONS: Quantitative measures derived from the occipital EEG are sensitive to AEDs and correlate with AED-related cognitive effects and subjective complaints. Although this correlation does not indicate a direct relation, quantified EEG may be a practical measure of AED impact on the brain. PMID- 12887434 TI - Induction of prolonged electrographic seizures in vitro has a defined threshold and is all or none: implications for diagnosis of status epilepticus. AB - PURPOSE: To study whether induction of prolonged (>30-min duration) in vitro electrographic seizure discharges resembling status epilepticus (SE) is graded or all-or-none, and to determine the critical factors mediating SE induction. METHODS: Prolonged electrographic seizure discharges were induced in combined hippocampal-entorhinal cortical (HEC) brain slices by electrical stimulation of the Schaeffer collaterals. Discharges were recorded by using field-potential electrodes in the dentate gyrus, CA3, CA1, and entorhinal cortex. Slices were prepared from rats that were (a). 21- to 30-day-old naive, (b). 60- to 120-day old naive, (c). epileptic, and (d). status post a prior traumatic brain injury. RESULTS: Induction of SE discharges was dependent on the duration, but not amplitude of the preceding stimulus train-induced afterdischarge in HEC slices from 21- to 30-day-old control, brain-injured, and epileptic animals, but not from 60- to 120-day-old animals. In slices from 21- to 30-day-old control animals, once afterdischarges exceeded 4 min in duration, SE was induced in 50% of slices, and after >or=6 min 37 s seizure activity; SE was induced in 95% of slices. A defined SE threshold also was evident in brain-damaged rats, including rats in which an epileptic condition was induced by pilocarpine injection 4-16 weeks before recording, and rats subjected to a fluid percussive head trauma 1-8 weeks before recording. However, in these brain-damaged animals, mean SE threshold was considerably lower (24 and 44 s, respectively). HEC slices from 60- to 120-day-old controls for the brain-injured and epileptic animals did not develop SE even after 20 stimulations, demonstrating the pronounced effect of brain injury and epilepsy on the development of SE in the HEC slice preparation compared with that in age-matched controls. CONCLUSIONS: In vitro, SE discharges have a defined temporal threshold for initiation. Once a seizure exceeds 6-7 min in duration in control animals, and 30-55 s in brain-damaged animals, the probability of SE induction is greatly increased. This demonstrates that brain injury lowers the afterdischarge duration required to produce SE and suggests that brains injured from trauma or SE are more susceptible to develop status epilepticus. PMID- 12887436 TI - Angelman syndrome: difficulties in EEG pattern recognition and possible misinterpretations. AB - PURPOSE: This study aimed to evaluate the sensitivity of the EEG in Angelman syndrome (AS), to verify the age at onset of suggestive EEGs and to study EEG patterns, analyzing variations and comparing our findings with nomenclature previously used. METHODS: Seventy EEG and 15 V-EEGs of 26 patients were analyzed. Suggestive EEG patterns of AS were classified in delta pattern (DP), theta pattern (TP), and posterior discharges (PDs). Generic terms were used to simplify the analysis. RESULTS: Suggestive EEGs were observed in 25 (96.2%) patients. DP occurred in 22 patients with four variants-hypsarrhythmic-like: irregular, high amplitude, generalized delta activity (DA) with multifocal epileptiform discharges (EDs); slow variant: regular, high-amplitude, generalized DA with rare EDs; ill-defined slow spike-and-wave: regular, high-amplitude, generalized DA with superimposed EDs characterizing a slow wave, with notched appearance; triphasic-like: rhythmic, moderate-amplitude DA over anterior regions with superimposed EDs. TP was observed in eight patients, as generalized or over the posterior regions. PDs were seen in 19 patients as runs of sharp waves or runs of high-amplitude slow waves with superimposed EDs. TP was the only age-related pattern (younger than 8 years) and observed only in patients with deletion. In 15 patients who had an EEG before the clinical diagnosis, 60% had a suggestive tracing. CONCLUSIONS: Although some EEG descriptions are not very detailed, and every author describes findings in a slightly different manner, obviously a common denominator must exist. In this context, EEG seems to be a very sensitive method for the diagnosis of AS, offering an opportunity to corroborate this etiologic diagnosis. Conversely, we do not believe that these patterns may be accounted as specific, except for the delta pattern, which seems to be extremely unusual in other syndromes. Other EEG patterns observed in AS, such as theta activity and PDs, occur in a wide variety of disorders. Nonetheless, their importance for the EEG diagnosis of AS is supported by the fact that they are associated with other features and may be helpful in a proper clinical setting. PMID- 12887437 TI - Ictal spitting: clinical and electroencephalographic features. AB - PURPOSE: To identify clinical and EEG correlates of ictal spitting automatisms and to assess their reliability in indicating the hemisphere of seizure onset. METHODS: The epilepsy-monitoring database (1994-2002) of the Cleveland Clinic Foundation (CCF) was searched for patients with a definite history of ictal spitting. All available documents of the patients, particularly their original video and EEG data, were reviewed. RESULTS: Twelve (0.3%) of the approximately 4000 patients had a documented history of ictal spitting. In seven of them, 15 seizures with spitting automatisms were recorded. All of them started with an aura or arousal out of sleep. In six of the seven patients (12 of 15 seizures), EEG onset was clearly lateralized to the right, nondominant hemisphere. Spitting occurred at a median time of 21 s after EEG seizure onset. At that time, predominantly fast, high-amplitude theta (5-7 Hz) was seen in the hemisphere of seizure onset, maximum temporal. In all but one of the total 12 patients, the epileptogenic zone was in the temporal lobe. In nine of the 12 patients, seizure onset was in the non-language-dominant hemisphere. Two patients had seizures arising from the language-dominant hemisphere; in another patient, the side of the seizure onset could not be determined. CONCLUSIONS: Ictal spitting is an uncommon feature of epileptic seizures. Although the symptomatogenic area is probably outside the temporal lobe, it is most frequently seen in temporal lobe epilepsy of the right, nondominant hemisphere. PMID- 12887438 TI - Entorhinal cortex MRI assessment in temporal, extratemporal, and idiopathic generalized epilepsy. AB - PURPOSE: We previously showed a reduction in the volume of the entorhinal cortex (EC) ipsilateral to the seizure focus in patients with intractable temporal lobe epilepsy (TLE). The purpose of this study was to examine the specificity of EC atrophy in epilepsy. METHODS: We performed volumetric measurement of the EC on high-resolution magnetic resonance imaging (MRI) in patients with TLE (n = 70), extratemporal lobe epilepsy (ETE; n = 18), and idiopathic generalized epilepsy (IGE; n = 20). EC volumes of epilepsy patients were compared with those of 48 age and sex-matched normal controls. Within the TLE group, 63 patients were selected prospectively with hippocampal atrophy ipsilateral to the seizure focus. The remaining seven patients were chosen retrospectively based on normal volumetric MRI of the hippocampus and amygdale, as well as normal histopathologic examination of the resected tissue. RESULTS: Compared with normal controls, EC volume was smaller ipsilateral but not contralateral to the seizure focus in patients with TLE (p < 0.001). No difference in the EC volumes ipsilateral and contralateral to the seizure focus was seen in patients with ETE and IGE compared with normal controls. The individual analysis showed that the EC was atrophic in 73% of TLE patients with hippocampal atrophy. Three of the seven TLE patients with normal volumetric MRI of the hippocampus and amygdala and normal histopathologic examination had EC atrophy ipsilateral to the seizure focus. In no patient with ETE or IGE was the EC found to be atrophic. CONCLUSIONS: EC atrophy ipsilateral to the seizure focus appears to be specific to mesial temporal lobe structural damage associated with TLE. PMID- 12887439 TI - Identifying potential surgical candidates in patients with evidence of bitemporal epilepsy. AB - PURPOSE: To determine which patients with evidence of medically refractory bitemporal epilepsy are potentially good candidates for surgical therapy. METHODS: We reviewed 42 adults with intractable seizures who were found to have bitemporal ictal onsets, based on scalp video-EEG long-term monitoring (LTM). All underwent invasive LTM before surgery. Surgical outcomes were classified as seizure free, >75% reduction in seizures, or <75% reduction in seizures, >or=1 year after resection. We related the following factors to outcome: (a). >75% preponderance of interictal scalp EEG discharges to one temporal region; (b). magnetic resonance imaging (MRI) findings; (c). lateralizing deficits on verbal or visual reproduction memory testing; and (d). memory failure with injection contralateral to side of surgery on Wada testing. RESULTS: Twenty-six (62%) of 42 patients had unilateral ictal onsets based on intracranial studies. Seizure freedom (occurring in 64% of this group), or >75% seizure reduction (found in 12% of subjects) occurred only when at least one of the following three factors was concordant with the side of surgery: preponderance of interictal scalp EEG discharges, unilateral temporal lesion on MRI, or lateralizing verbal or visual reproduction memory deficits on neuropsychological tests (p = 0.004). Seven subjects with bilateral ictal onsets based on intracranial studies had resections based on preponderance of seizures to one side, or other lateralizing noninvasive abnormality. Five of these (all of whom had >or=80% of seizures originating from one side) had >75% reduction in seizures. CONCLUSIONS: Invasive monitoring to pursue possible surgical therapy for patients with surface EEG evidence of bitemporal epilepsy may be justified only when some lateralizing feature is found in other noninvasive assessments. PMID- 12887440 TI - Outcome of surgical treatment in familial mesial temporal lobe epilepsy. AB - PURPOSE: To describe postoperative outcome in patients with familial mesial temporal lobe epilepsy (FMTLE). METHODS: We studied FMTLE patients who underwent surgical treatment for refractory seizures. FMTLE was defined when at least two individuals in a family had a clinical EEG diagnosis of MTLE. Preoperative investigation included magnetic resonance imaging (MRI), interictal/ictal EEGs, and neuropsychological evaluation. We used Engel's classification for postoperative outcome. RESULTS: To date, 20 FMTLE patients have been operated on, with 1.6 to 9.8 years of follow-up (mean, 5.5 years). Hippocampal atrophy (HA) and other signs of mesial temporal sclerosis (MTS) were present in 18 patients (15 unilateral). Seizures were recorded in 19 patients. Seventeen (85%) patients are in class I. Two patients had normal hippocampal volumes (HcV): one (5%) is in class II and the other (5%) in class IV (extratemporal seizures developed after surgery). One (5%) patient had bilateral HA and is in class III. Qualitative histopathology showed MTS with different degrees of severity. CONCLUSIONS: Refractory FMTLE patients have good surgical outcome when unilateral or clearly asymmetric HA is identified. Preoperative investigation should be the same as that in patients with sporadic refractory MTLE. PMID- 12887441 TI - Short-term nonhormonal and nonsteroid treatment in West syndrome. AB - PURPOSE: West syndrome (WS) is considered an age-dependent epileptic encephalopathy and also a particular type of electrical epileptic status. Short term hormonal or steroid treatment of WS with good efficacy is reported in the literature. The aim of this retrospective multiinstitutional study was to evaluate the early discontinuation of nonhormonal and nonsteroid treatment for WS. METHODS: Twenty-two WS cases in which treatment was discontinued after a maximum of 6 months, were collected. Inclusion criteria were the presence of typical EEG hypsarrhythmia (HY) and video-EEG recorded epileptic spasms. Exclusion criteria were the presence of partial seizures or other seizure types before spasm onset. The patients were treated with vigabatrin (VGB) in 19 cases and nitrazepam (NTZ) in three. The dose range was 70-130 mg/kg/day for VGB and 0.7-1.5 mg/kg/day for NTZ. The drug was discontinued if spasms stopped and HY disappeared after a mean treatment period of 5.1 months (range, 3-6 months). All patients underwent repeated and prolonged awake and sleep video-EEG, both before and after drug discontinuation. RESULTS: Cryptogenic (15) and symptomatic (seven) WS patients were included. All the symptomatic cases had neonatal hypoxic ischemic encephalopathy. The mean age at spasm onset was 5.5 months (range, 3-7 months; median, 6). The interval between spasm onset and drug administration ranged from 7 to 90 days (mean, 23 days; median, 20). The interval between drug administration and spasm disappearance ranged from 2 to 11 days (mean, 6 days; median, 6 days). The interval between drug administration and HY disappearance ranged from 3 to 30 days (mean, 9 days; median, 10 days). Drugs were stopped progressively over a 30- to 60-day period. Follow-up ranged from 13 to 50 months (mean, 26 months; median, 22 months). None of our cases showed spasm recurrence. CONCLUSIONS: Our data show that successful nonhormonal and nonsteroid treatment can be shortened to a few months without spasm recurrence in patients with cryptogenic or postanoxic WS. PMID- 12887442 TI - Association of the neuronal nicotinic acetylcholine receptor subunit alpha4 polymorphisms with febrile convulsions. AB - PURPOSE: The alpha4-subunit gene of the neuronal nicotinic acetylcholine receptor (CHRNA4) has been identified as the first gene underlying an idiopathic partial epilepsy syndrome in human autosomal-dominant nocturnal frontal lobe epilepsy. Studies provided evidence that the protein coded by CHRNA4 is one of the most abundant subunits of the neuronal nicotinic acetylcholine receptors in mammalian brains, and mutations of CHRNA4 seem to cause neuronal excitation. The CHRNA4 gene may have a role in the development of febrile convulsions (FCs), the majority of childhood seizures. This study assessed the distribution of genotypes of CHRNA4 in patients with FCs. METHODS: A total of 102 children with FCs and 80 normal control subjects were included in the study. Polymerase chain reaction was used to identify the C/T polymorphism of the CHRNA4 gene. Genotypes and allelic frequencies for the CHRNA4 gene polymorphisms in both groups were compared. RESULTS: The number of individuals with heterozygous CHRNA4 (Ser543Ser)-C/T genotype was significantly greater (60.8% vs. 32.5%; p = 0.001), and the CHRNA4 (Ser543Ser)-T allele frequency was significantly higher (p = 0.001), in patients with FCs compared with healthy controls. The odds ratio for developing FCs in individuals with the CHRNA4 (Ser543Ser)-CT genotype was 3.77 compared with individuals with two copies of the CHRNA4 (Ser543Ser)-C allele. CONCLUSIONS: This study demonstrated an association between the CHRNA4 gene and FCs. Individuals with the T allele had a higher incidence of FCs. These data suggest that the CHRNA4 gene or a closely linked gene might be one of the susceptibility factors for FCs. PMID- 12887443 TI - Expressed emotion in the relatives of people with epileptic or nonepileptic seizures. AB - PURPOSE: This study investigated Expressed Emotion (EE) in relatives of people with epileptic or nonepileptic seizures (NES). METHODS: In a cross-sectional study, we used the Five-Minute Speech Sample to explore EE in the key relative of people with epilepsy (n = 36) and those with NESs (n = 21), as well as levels of anxiety and depression and use of coping strategies. RESULTS: A significantly greater proportion of relatives of NES than epilepsy patients were rated as high EE. Hostility was evident in more high-EE epilepsy than high-EE NES relatives, whereas emotional overinvolvement and positive relationship ratings tended to be more common in high-EE NES relatives. High- and low-EE epilepsy relatives used problem-focused as opposed to emotion-focused coping strategies significantly more than half the time. High EE and seizure frequency were not associated. Age at onset of the disorder was higher in epilepsy patients with high- than with low EE relatives. CONCLUSIONS: Irrespective of etiology, carers for people with seizure disorders may find it hard to adjust to the difficulties these disorders create. Interventions that encourage problem-solving, reappraisals of "loss" and education regarding the causes of some of the patients' behavioral and mood problems seem likely to be beneficial. PMID- 12887444 TI - Dental status and oral health of patients with epilepsy: an epidemiologic study. AB - PURPOSE: We performed a dental survey of epilepsy patients to examine their oral health by statistical means and to provide a guide for the dental treatment of these patients. METHODS: We first set up four "dental" subgroups of epilepsy patients, based on the types of seizures, seizure frequency, and mental state. One hundred one patients underwent a survey concerning their dental, medical, and epilepsy histories, followed by a dental examination. Indexes quantifying oral hygiene, the number and condition of the remaining teeth, periodontium, and the degree of prosthetic treatment were measured. An age-matched control group of general (nonepilepsy) population underwent an identical dental examination. Statistical comparison was performed between the patient and the control groups and between subgroups of epilepsy patients. RESULTS: In almost all aspects of oral health and dental status, patients with epilepsy showed a significantly worse condition compared with the control group. Comparison of the subgroups of epilepsy patients revealed that the most severe findings concern patients who have poorly controlled epilepsy, especially those who have frequent generalized tonic-clonic seizures. CONCLUSIONS: The observed difference probably results from a combination of factors such as the effect of the seizures themselves, socioeconomic conditions, and the negative attitude of dentists. We recommend that the planning of dental treatment of such patients should start with the assessment of their disease and determination of the "dental" subgroup to which they belong. For each subgroup, specific recommendations for interventions are given. PMID- 12887445 TI - Diazepam terminates brief but not prolonged seizures in young, naive rats. AB - PURPOSE: Ample evidence exists from both clinical and animal studies that the success of benzodiazepine intervention during status epilepticus (SE) in the mature nervous system is inversely related to seizure duration. This relationship has not been well studied in the developing nervous system. METHODS: The objective of this study was to investigate the relation of age and success of diazepam (DZP) treatment in the lithium-pilocarpine model of secondarily generalized seizure in the rat by using naive rats of three age groups, roughly corresponding to the human ages of infancy (P15), adolescence (P20), and adult (P60). RESULTS: In all age groups, the dosage of DZP that stopped the seizures at 5 min was not effective in terminating seizures at 60 min. This decline in efficacy was present as early as 15 min after seizure onset. CONCLUSIONS: These findings demonstrate that the inverse relation between the success of benzodiazepine intervention and seizure duration is observed in young as well as in adult rats and provide further evidence that intervention for SE should commence early. PMID- 12887446 TI - Evidence for S284L mutation of the CHRNA4 in a white family with autosomal dominant nocturnal frontal lobe epilepsy. AB - PURPOSE: To identify mutations of the neuronal nicotinic acetylcholine receptor alpha4 subunit gene (CHRNA4) responsible for autosomal dominant nocturnal frontal lobe epilepsy (ADNFLE) in a group of white patients. METHODS: A group of 47 patients from 21 unrelated families with ADNFLE were screened for mutations in CHRNA4. Clinical features and EEG findings in the patients were consistent with those reported in the literature for other affected families. The entire gene was amplified from genomic DNA by polymerase chain reaction (PCR) followed by multitemperature single-strand conformation polymorphism analysis (MSSCP) and sequencing. RESULTS: A c.851C>T transition in exon 5 of CHRNA4 was identified in three affected individuals from two generations of the same family, but not in the remaining patients or in 100 healthy volunteers. This mutation caused an S284L substitution in the transmembrane domain M2 segment of the alpha4 subunit of the neuronal nicotinic acetylcholine receptor. The same mutation had previously been detected in a single Japanese family with ADNFLE, and in an Australian woman with a sporadic form of NFLE. CONCLUSIONS: This is the first report of an occurrence of c.851C>T transition in a white family with ADNFLE. PMID- 12887447 TI - Rasmussen syndrome: multifocal spread of inflammation suggested from MRI and PET findings. AB - BACKGROUND: A 6-year-old girl with Rasmussen syndrome (RS) showed multiple small high-signal-intensity areas independently in the right hemisphere by fluid attenuated inversion recovery (FLAIR) imaging on magnetic resonance imaging (MRI) 1 year after the onset of epilepsy. METHODS: MRI performed 4 months later demonstrated a further increase in the number of these foci and enlargement in the size of the previous FLAIR lesions. RESULTS: An [18F]-fluorodeoxyglucose positron emission tomography (FDG-PET) study showed a strong, spotty uptake in the right temporooccipital regions, corresponding to the sites of continuous EEG seizure discharges. In contrast, [11C]methionine PET demonstrated multifocal uptake regions, which corresponded anatomically to the FLAIR lesions, suggesting sites of underlying chronic inflammation. CONCLUSIONS: These neuroimaging findings suggested that the inflammatory process in RS spreads either multifocally at the same time, as seen in this case, or from one discrete area to the adjacent region, as reported previously. PMID- 12887448 TI - Confirmation of nonconvulsive limbic status epilepticus with the sodium amytal test. AB - Is it a seizure? This question can be difficult for a clinician to answer, and it may be more critical if the possible seizure lasts >30 min. Long-duration questionable seizure activity changes the question to, "Is it status epilepticus?" Status epilepticus (SE) can be divided into convulsive and nonconvulsive types. Convulsive SE is the most easily recognized, whereas nonconvulsive SE is more clinically variable and controversial. The term nonconvulsive SE is more often applied to patients who are severely obtunded or comatose with minimal or no motor movements, or in a stupor of altered consciousness reflecting generalized ictal activity. Nonconvulsive SE also can be caused by focal seizure activity, sometimes restricted to deep small volumes of brain in which scalp EEG may not be diagnostic. We present the case of a patient who had dominant limbic hippocampal SE, but in whom the diagnosis could not be confirmed until a modified novel use of the sodium amytal test was performed. PMID- 12887449 TI - Famine-associated AED toxicity in rural Zambia. PMID- 12887450 TI - Abnormal ALP isoenzyme in children with epilepsy treated with carbamazepine. PMID- 12887462 TI - Prostate-specific antigen: should patients receive information before having the test? PMID- 12887463 TI - Clinical trials in prostate cancer. PMID- 12887464 TI - Surgical factors in bladder cancer: more (nodes) + more (pathology) = less (mortality). PMID- 12887465 TI - Metallic ureteric stents: the current situation. PMID- 12887466 TI - Screening for prostate cancer: have you had your cholesterol measured? AB - Screening for prostate cancer has become one of the most common topics of conversation at urological oncology meetings. Most people have a bias as to whether there should or should not be a national screening programme. Unfortunately there are many unanswered questions, which may or may not be possible to answer definitively. In a balanced and scholarly review of the subject, Professor Peter Boyle indicates several flaws in the agreement for screening, but feels that PSA testing will continue unabated. The authors from the University of Stellenbosch review the plentiful literature relating to testicular torsion and functional recovery. They also review the mechanism of injury and the effect on the contralateral testis. PMID- 12887467 TI - Testicular function after torsion of the spermatic cord. PMID- 12887468 TI - A prospective comparison of radical retropubic and robot-assisted prostatectomy: experience in one institution. AB - The authors from the Vattikuti Institute in the USA report a prospective comparison of radical prostatectomy and robot-assisted prostatectomy. They found that the robot-assisted procedure was safer, and yielded favourable oncological and functional results. They also present work in association with the Department of Urology in Mansoura into robot-assisted radical cystoprostatectomy and urinary diversion, and point out the advantages and disadvantages associated with performing the most complex types of urinary diversion. There is also an interesting paper relating to the association between sexual factors and prostate cancer, from authors in institutions in Australia, New Zealand and Italy. They found that in a case-control study of men aged <70 years, ejaculatory frequency was negatively associated with the risk of prostate cancer. Technology has made many contributions to the management of urological patients. The classic example is that of urinary stone management. Authors from the USA evaluated cyroablation of renal carcinoma in patients with solitary kidneys. They are encouraged by their results and suggest that there is merit in this treatment, but indicate the need for a longer follow-up. OBJECTIVE: To prospectively compare standard radical retropubic prostatectomy (RRP) and the robotically assisted Vattikuti Institute prostatectomy (VIP) in the management of localized prostate cancer. PATIENTS AND METHODS: The study was a single-institution, prospective, unrandomized comparison of histopathological, and functional outcomes, at baseline and during and after surgery, in 100 patients undergoing RRP and 200 undergoing VIP. RESULTS: While the variables before surgery, the operative duration (163 vs 160 min) and pathological stages were comparable, there were significant differences in the measured outcomes. The blood loss was 910 and 150 mL for RRP and VIP, respectively, and transfusion was greater after RRP (67% vs none; both P < 0.001). There were four times as many complications after RRP (20% vs 5%, P < 0.05), the haemoglobin level at discharge was lower (100 vs 130 g/L, P < 0.005) and the hospital stay longer (3.5 vs 1.2 days; P < 0.05). Most (93%) of VIP and none of the RRP patients were discharged within 24 h (P < 0.001); the duration of catheterization was twice as long after RRP (15.8 vs 7 days; P < 0.05). Positive margin was more frequent after RRP (23% vs 9%, P < 0.05). After VIP, patients achieved continence and return of erections more quickly than after RRP (160 vs 44, and 180 vs 440 days, both P < 0.5). The median return to intercourse was 340 days after VIP but after RRP half the patients have as yet not resumed intercourse at 700 days (P < 0.05). CONCLUSIONS: The VIP procedure appears to be safer, less bloody and requires shorter hospitalization and catheterization. The oncological and functional results were favourable in patients undergoing VIP. PMID- 12887469 TI - Sexual factors and prostate cancer. AB - OBJECTIVE: To assess whether prostate cancer might be related to hormone levels and, by inference, to differences in sexual activity. PATIENTS, SUBJECTS AND METHODS: In a case-control study of men with prostate cancer aged < 70 years at diagnosis and age-matched control subjects, information was collected on two aspects of sexual activity; the number of sexual partners and the frequency of total ejaculations during the third to fifth decades of life. RESULTS: There was no association of prostate cancer with the number of sexual partners or with the maximum number of ejaculations in 24 h. There was a negative trend (P < 0.01) for the association between risk and number of ejaculations in the third decade, independent of those in the fourth or fifth. Men who averaged five or more ejaculations weekly in their 20s had an odds ratio (95% confidence interval) of 0.66 (0.49-0.87) compared with those who ejaculated less often. CONCLUSIONS: The null association with the number of sexual partners argues against infection as a cause of prostate cancer in this population. Ejaculatory frequency, especially in early adult life, is negatively associated with the risk of prostate cancer, and thus the molecular biological consequences of suppressed or diminished ejaculation are worthy of further research. PMID- 12887470 TI - Health-related quality of life and psychosocial factors in patients with prostate cancer scheduled for radical prostatectomy or external radiation therapy. AB - OBJECTIVE: To assess whether baseline health-related quality of life (HRQOL) and psychosocial profiles differ in patients with prostate cancer scheduled for radical prostatectomy (RP) or external radiation therapy (ERT), as there is evidence that HRQOL is influenced by psychosocial factors (PFs), so that any variation at baseline should be considered when comparing the effect of therapy on HRQOL. PATIENTS AND METHODS: Before receiving therapy, HRQOL and PFs were assessed in 65 patients scheduled for RP and in 73 scheduled for ERT. To measure HRQOL (generic and disease-specific) and PFs, an extended questionnaire was constructed, using validated and standardized instruments. Clinical data were collected from patients' medical records. Comparisons adjusted for age and socio economic status (SES) were analysed using Student's t-test and univariate analyses of variance and covariance. RESULTS: Patients scheduled for ERT were 7.9 years older and had a lower SES (both P < 0.001), more often had stage T3 and T4 disease, had poorer histopathological differentiation and higher levels of prostate-specific antigen (all P < 0.01). They also reported a worse physical, role, cognitive and social function, more fatigue, more pain, a lower overall HRQOL and worse sexual function than patients scheduled for RP. There were no differences in urinary and bowel function, nor in the PFs assessed. CONCLUSION: The baseline HRQOL profile of patients scheduled for RP is better than in those scheduled for ERT. These results are in line with those from the few other studies on this subject. Knowing the impact of RP and ERT on HRQOL should therefore be based mainly on longitudinal studies including baseline measures, the analyses of which should be adjusted for age and SES. In the present small study, baseline PFs did not differ between the treatments. PMID- 12887471 TI - Putative protein markers in the sera of men with prostatic neoplasms. AB - OBJECTIVE: To describe the preliminary identification of serum proteins that may be diagnostic markers in prostate cancer. PATIENTS AND METHODS: The study included 11 men referred for treatment of localized prostate cancer, 12 with benign prostatic hyperplasia (BPH) and 12 disease-free controls. For serum protein analysis, the protein-chip array surface-enhanced laser desorption/ionization (SELDI) technique was used (Ciphergen Biosystems, Fremont, CA). SELDI combines protein-chip technology with time-of-flight mass spectrometry, and offers the advantages of speed, simplicity and sensitivity. RESULTS: Three protein peaks were identified in the serum of men with prostate cancer and BPH, but not in controls, with relative molecular masses of 15.2, 15.9 and 17.5 kDa. These three proteins were significantly associated with BPH and prostate cancer when compared with controls (P = 0.001, 0.004, and 0.011, respectively, Kruskal-Wallis test). Interestingly, the 17.5 kDa protein was more abundant in five men with stage T1 prostate cancer than in eight with stage T2 (P = 0.016, two tailed Mann-Whitney U-test corrected for ties). CONCLUSIONS: These proteins, particularly the 15.9 kDa one, may be used for the diagnosis or monitoring of prostate cancer and differentiation from BPH, and have the potential for antibody-based chip SELDI-TOF technology. Identified proteins may be targets for immunotherapy. PMID- 12887472 TI - Comparative efficacy of triptorelin pamoate and leuprolide acetate in men with advanced prostate cancer. AB - OBJECTIVE: To compare the efficacy of monthly administrations of the luteinizing hormone-releasing hormone agonists triptorelin pamoate and leuprolide acetate to induce and maintain castrate levels of serum testosterone in men with advanced prostate cancer. PATIENTS AND METHODS: Men with advanced prostate cancer were randomly assigned to receive triptorelin 3.75 mg or leuprolide 7.5 mg. The agent was injected intramuscularly every 28 days for nine injections. Primary endpoints were the percentages of men whose serum testosterone concentrations declined to and were maintained at or below castrate levels (/= 50 years with moderate to severe symptoms (American Urological Symptom Index, AUA-SI, score >/= 12), a prostate volume of >/= 30 mL, a serum prostate-specific antigen (PSA) level of >/= 1.5 or < 10 ng/mL, and a maximum urinary flow rate (Qmax) of /= 5 (greatest symptomatic burden) treatment with dutasteride improved the scores by 2.41, while the scores in placebo-treated patients only improved by 1.64. Dutasteride-treated patients with a baseline BII score of < 5 (least symptom burden) had a clinically significant improvement in health status, while placebo-treated patients deteriorated. Regression analysis showed that men with a combination of a baseline BII item-3 score of 3 (bothered a lot) and a high symptom score (AUA-SI >/= 20) were more likely to be bothered by their symptoms at the end of the study. Men receiving placebo were also more likely to be bothered at the end of the study than were those receiving dutasteride. CONCLUSIONS: Dutasteride treatment is associated with clinically significant improvements in BII score, reflecting improvements in the quality of life of men with BPH. Taken together with previously reported improvements in prostate volume, lower urinary tract symptoms and urinary flow, and diminution of the risk of acute urinary retention and the need for BPH related surgery, dutasteride offers demonstrable efficacy in the management of BPH. PMID- 12887481 TI - Serenoa repens extract for benign prostate hyperplasia: a randomized controlled trial. AB - OBJECTIVE: To compare the effect of a Serenoa repens extract with placebo for symptoms of benign prostatic hyperplasia (BPH). PATIENTS AND METHODS: In a double blind placebo-controlled randomized trial between January 1999 and March 2000, 100 men with symptoms of BPH, aged < 80 years, with a maximum urinary flow rate of 5-15 mL/s for a voiding volume of 150 mL, were randomly and equally allocated to 320 mg S. repens extract or placebo (paraffin oil). The main outcome measures were the International Prostate Symptom Score (IPSS), peak urinary flow rate, and the Rosen International Index of Erectile Function (IIEF) questionnaire. RESULTS: There was no significant difference between the treatments over the 12 weeks of the study in the IPSS, peak urinary flow rate or for the IIEF questionnaire. CONCLUSIONS: During the trial all participants had some improvement in their symptoms of BPH but there was no significant beneficial effect of this S. repens extract over placebo in this 12-week trial. PMID- 12887482 TI - Continent urinary tract reconstruction - the Lund experience. AB - The Department of Urology in Lund, Sweden, has a long association with innovations in reconstructive urology. The authors from that department describe their experience over a long period with orthotopic bladder substitution and continent cutaneous urinary diversion. They conclude that continent urinary tract reconstruction is associated with a high incidence of early and late complications. They also found that for storage and emptying, their Lundiana pouch was superior to the Goldwasser neobladder. OBJECTIVE: To assess the early and late complications and functional results in patients undergoing continent reconstruction of the urinary tract, i.e. orthotopic bladder substitution (OBS) or continent cutaneous diversion (CCD). PATIENTS AND METHODS: The medical records of all patients undergoing OBS (Goldwasser technique) or CCD ('Lundiana' technique) for malignant or benign disease during 1987-1999 and followed to December 2001 were reviewed. There were 67 patients with neobladders, 77 with a Lundiana pouch who had undergone radical cystectomy and 22 with a Lundiana pouch operated for benign disorders. RESULTS: Early complications requiring reoperation occurred in 12% of the cystectomy group, with no difference with type of reconstruction, and in 10% with benign diseases. Four patients (3%) undergoing radical cystectomy died from early cardiovascular complications, two after surgery for intra-abdominal complications. Intestinally related complications and wound dehiscence requiring re-operation occurred in nine and six patients, respectively. The incidence of late complications requiring open surgery was 22% and 23% after cystectomy with OBS and CCD, respectively. The value in patients with benign diseases undergoing CCD was also 23%. Stone formation in the pouch was common, occurring in 12% in patients with OBS and in 10% after CCD. The pouch perforated or ruptured in four patients. The incidence of uretero-intestinal stricture using the Le Duc technique was 2.4% and renal function was well preserved. The incidence of revisional surgery of the Lundiana pouch outlet for incontinence was low and all patients but four were continent. The functional outcome in patients with OBS was less good; some needed pouch augmentation or an artificial urinary sphincter. Most patients used incontinence products and many needed clean intermittent self-catheterization. CONCLUSION: Continent urinary tract reconstruction is associated with a high incidence of early and late complications. For storage and emptying, the CCD Lundiana pouch is superior to the OBS of Goldwasser. PMID- 12887483 TI - Preference for oral sildenafil or intracavernosal injection in patients with erectile dysfunction already using intracavernosal injection for > 1 year. AB - Authors from Seoul describe their experience with patients already on triple therapy by intracavernosal injection who changed to oral sildenafil. Rather surprisingly, they found that patients had had a greater preference than expected for triple therapy, feeling that they had a better quality of erection on intracavernosal injection. The subject of the effect of renal transplantation on sperm quality and sex hormone levels is discussed by authors from Teheran. They found that sperm morphology and density remained unchanged, but there were significant improvements in sperm mobility. There was also an improvement in hormone levels and sexual function. OBJECTIVE: To investigate the efficacy and preference for oral sildenafil or intracavernosal injection (ICI) therapy in patients with erectile dysfunction (ED) already using ICI. PATIENTS AND METHODS: In all, 69 patients with ED (mean age 55.1 years, sd 12.3) on ICI therapy with triple solution (papaverine/phentolamine/prostaglandin-E1) for > 1 year were recruited for the study. Their erection quality, adverse reactions and selection rate of oral sildenafil or ICI as treatment, after using sildenafil for 3 months, and the reasons for their preferences, were compared between the regimens, RESULTS: Overall, 52 men (75%) responded to sildenafil; of these men, the erection quality with ICI was better than that with sildenafil in 46 (89%) and 16 (31%) preferred ICI as their treatment. Eighteen patients (35%) used each treatment alternately and 18 (35%) used sildenafil exclusively. The main reason given by patients for choosing ICI was a better quality of erection (74%). CONCLUSION: More patients with ED and using ICI preferred it as their main treatment than was expected, even though they had a good response to oral sildenafil. A better quality of erection with ICI was the reason why experienced patients chose this method, differing from the choice of patients starting treatment for ED. PMID- 12887484 TI - Effect of renal transplantation on sperm quality and sex hormone levels. AB - OBJECTIVE: To assess the effect of successful renal transplantation on semen variables, sexual function and sex hormone profiles in a clinical trial. PATIENTS AND METHODS: Thirty patients on haemodialysis underwent renal transplantation; before and after surgery, their sperm density, motility and morphology were analysed, follicle-stimulating hormone (FSH), luteinizing hormone (LH), prolactin and testosterone levels measured and compared, and sexual function assessed using an abbreviated version of the International Index of Erectile Function (IIEF), with a successful outcome defined as a level of satisfaction of 4 or 5 on a 5 point scale. The paired t-test was used to assess the statistically significance of differences in all analyses. RESULTS: Sperm motility improved significantly (P < 0.001) but there were no significant changes in morphology or density (P = 0.33 and 0.068, respectively). Testosterone levels increased and FSH, LH and prolactin decreased significantly (P < 0.05) after renal transplantation. The IIEF showed that of the 30 patients, 14 were impotent before surgery and only six remained so afterward (P < 0.05). CONCLUSION: Although sperm morphology and density did not improve after renal transplantation, there were highly significant changes in sperm motility. Hormonal levels in patients on haemodialysis improved after transplantation and returned to nearly normal; sexual function was also significantly better. Further studies are needed to confirm these results. PMID- 12887485 TI - Treatment of vesico-ureteric reflux: a new algorithm based on parental preference. AB - Authors from Rome evaluated parental preference for treatment in children with grade III VUR. Parents were provided with detailed information about the three treatment options: antibiotic treatment, open surgery, endoscopic treatment. Most parents chose endoscopic management; with this in mind, the authors proposed a new treatment algorithm for VUR. OBJECTIVE: To assess parental preference (acknowledged in treatment guidelines as important when choosing therapy) about treatments for vesico-ureteric reflux (VUR, commonly associated with urinary tract infection and which can cause long-term renal damage if left untreated), as at present there is no definitive treatment for VUR of moderate severity (grade III). SUBJECTS AND METHODS: The parents of 100 children with grade III reflux (38 boys and 62 girls, mean age 4 years, range 1-15) were provided with detailed information about the three treatment options available for treating VUR (antibiotic prophylaxis, open surgery and endoscopic treatment), including the mode of action, cure rate and possible complications, and the practical advantages and disadvantages. They were then presented with a questionnaire asking them to choose their preferred treatment. RESULTS: Most parents preferred endoscopic treatment (80%), rather than antibiotic prophylaxis (5%) or open surgery (2%); 13% could not decide among the three options and endoscopic treatment was recommended. CONCLUSION: Given the strong preference for endoscopic treatment we propose a new algorithm for treating VUR; endoscopic treatment would be considered as the first option for persistent VUR, except in severe cases where open surgery would still be recommended. PMID- 12887486 TI - Neurovesical dysfunction in children after treating pelvic neoplasms. AB - OBJECTIVE: To evaluate 10 years of experience, and thus define the occurrence and causes, of neurogenic lower urinary tract dysfunction in children with pelvic neoplasms treated by surgery. PATIENTS AND METHODS: From 1991 to 2000, 33 children were operated by the same surgeons for pelvic neoplasms; 11 were analysed, comprising four each with sacrococcygeal teratoma (ST) and ganglioneuroma, and one each with yolk sac tumour (YST), neuroblastoma and myofibroblastic bladder sarcoma (MBS). The other patients were not assessed because eight had died or were in severe progression, three were treated by bladder substitution and the others were lost to follow-up or refused a urological evaluation. All 11 children were evaluated at >/= 6 months after surgery with a questionnaire about bowel and voiding habits, a neurological and orthopaedic assessment, a noninvasive urodynamic study, renal ultrasonography and spinal and pelvic magnetic resonance imaging (MRI). All patients with signs of bladder dysfunction were evaluated by a pressure-flow study. The results were analysed for surgical approach and anatomical involvement, i.e. group A, extensive surgery for complete tumour excision in the sacral area (ST and YST); group B, surgery for tumour resection in the paraspinal ganglia area (neuroblastoma and ganglioneuroma); and group C, bladder tumour with partial bladder resection (MBS). RESULTS: Eight patients had signs or symptoms related to bladder sphincter dysfunction. One child refused the invasive urodynamic evaluation, leaving seven for analysis (two each ST and ganglioneuroma, one each YST, neuroblastoma and MBS). The urodynamic findings were normal in three children. On spinal and pelvic MRI a presacral lipoma with syringomyelia was discovered in one child with ST. Eight children had bladder dysfunction and two had no neurogenic damage (which was only in sacral tumours); in one child it was related to an upper motor neurone lesion from spinal dysraphism and in the other to a lower motor neurone lesion from surgical injury to the splanchnic nerves. Patients operated for paraspinal tumours had more bladder dysfunction but no signs of neurogenic damage, as did the patient with partial bladder resection. However in Group B, there may have been a transient or incomplete nerve injury in one patient. CONCLUSIONS: Deficits of parasympathetic, sympathetic and somatic innervation of the bladder and the urethra may occur in children after surgery for pelvic neoplasms, related to minor or major surgical trauma. In ST, a tethered cord may be associated with mixed neurogenic damage. Knowledge of bladder dysfunction in anorectal malformations, spinal dysraphism, etc. and the clinical protocol used in these patients also seemed to be useful for understanding the development of voiding dysfunction in patients with neoplasm. PMID- 12887487 TI - Previous testicular position in boys referred for an undescended testis: further explanation of the late orchidopexy enigma? AB - OBJECTIVE: To investigate the significance of the acquired undescended testis (UDT), which is differentiated into congenital and acquired forms, by assessing the previous testicular position in affected boys. PATIENTS AND METHODS: The study comprised 261 boys who had been referred for a non-scrotal testis to the outpatient clinic during an 8-year period (1993-2000). There was a bimodal distribution of age, with peaks at 2.0 and 10.0 years. In each boy with UDT the previous testicular position was ascertained. RESULTS: On referral, 340 testes were not in the scrotum (182 uni- and 79 bilateral). Of the 340 testes, 82 (24%) in 61 boys were diagnosed as retractile, whereas the remaining 258 in 221 boys were undescended. The previous testicular position was known in 208 of 221 boys (94%), with 244 UDTs. In 65 of these 244 (26.6%) the testis had never been scrotal (congenital UDT); in 179 (73.4%) a previous intrascrotal position was recorded in early childhood (acquired UDT) at least once, in 149 (61%) at least twice and in 117 (48%) at least three times. The mean age at referral for congenital UDT was 2.1 years and for acquired UDT was 8.4 years. CONCLUSIONS: These results show that acquired UDT is frequent, and occurs at about three times the rate of congenital UDT. Because these boys are referred for treatment later in childhood, the acquired UDT probably accounts for the high rate of (late) orchidopexy. PMID- 12887488 TI - Impedance measurements and connexin expression in human detrusor muscle from stable and unstable bladders. AB - Three of this month's Scientific Discovery papers highlight the importance of collaboration in delivering high quality scientific research. As scientific technology increases in power and cost, and specific areas of interest become more specialized, it is becoming more difficult to cover all aspects of a completeresearch story. Collaborating with other experts in the field or other fields, including industry, allows strong scientific proof to be generated for the hypothesis and aims. Building strong collaborative,inter-disciplinary, multi institutional, international groups with academic and industrial partners is the way forward for all discovery. We look forward to publishing more of these collaborative papersin future issues of the BJU International. OBJECTIVES: To test the hypothesis that intercellular electrical coupling is altered in human detrusor smooth muscle from patients with unstable bladders. MATERIALS AND METHODS: Human detrusor biopsy samples were obtained from patients with stable and unstable bladders. Intracellular electrical impedance was measured with alternating current (20 Hz-300 kHz) across the ends of detrusor strips in an oil gap, after correcting for extracellular space resistance. Gap junctions were identified by localization of connexins (Cx), specifically Cx45, Cx43 and Cx40 transcripts, using immunoconfocal microscopy. RESULTS: Total intracellular resistivity was greater in strips from unstable than from stable bladders (median 1246 vs 817 Omega.cm). The increase was attributed to an increase in junctional resistance; cytoplasmic resistance was unchanged. Cx43 was localized to a submucosal layer and to connective tissue; Cx40 label was confined to endothelial cells of blood vessels. Cx45 labelling was localized to detrusor bundles and appeared to be less marked in samples from unstable bladders. Semi-quantitative analysis of Northern blots showed that Cx45 expression in unstable was less than that in stable bladders. CONCLUSIONS: These data suggest that intercellular coupling is reduced in detrusor from unstable bladders. Cx45 was localized to the detrusor layer, with Cx 43 more evident in the suburothelial mucosa. Cx45 labelling was less intense in detrusor samples from unstable bladders. These results are consistent with reduced gap junction coupling in detrusor from unstable bladders. PMID- 12887489 TI - Skeletal growth and long-term bone turnover after enterocystoplasty in a chronic rat model. AB - OBJECTIVE: To investigate skeletal growth and bone metabolism in a chronic animal model of urinary diversion. MATERIALS AND METHODS: Young male Wistar rats (120) were allocated randomly to four groups undergoing: ileocystoplasty, ileocystoplasty and resection of the ileocaecal segment, colocystoplasty, and controls. All animals received antibiotics for 1 week after surgery; half of each group remained on oral antibiotics. Bone-related biochemistry was measured in serum and urine. Dual-energy X-ray absorptiometry and peripheral quantitative computed tomography (pQCT) were used to determine bone mass ex vivo. RESULTS: Most (90%) of the rats survived the study period (8 months); six rats died from bowel obstruction at the level of the entero-anastomosis and four had to be killed because of persistent severe diarrhoea. Vital intestinal mucosa was found in all augmented bladders. There were no differences in bone length and volume. Loss of bone mass was almost exclusively in rats with ileocystoplasty and resection of the ileocaecal segment (-37.5%, pQCT, P < 0.01). There was no hyperchloraemic metabolic acidosis or gross impairment of renal function. Hypomagnesaemia, hypocalcaemia and decreased insulin-like growth factor-binding protein 3 were the only significant findings on blood analysis. Deoxypyridinoline crosslinks in urine were higher in rats with an enterocystoplasty than in controls. CONCLUSIONS: Enterocystoplasty in rats neither impairs skeletal growth nor bone quantity, but leads to significant loss of bone mass when combined with resection of the ileocaecal segment. Rarefaction of the trabecular network is confined to the metabolically highly active cancellous compartment, most likely as a consequence of intestinal malabsorption. PMID- 12887490 TI - The growth inhibitory effect of p21 adenovirus on androgen-dependent and independent human prostate cancer cells. AB - OBJECTIVE: To assess the potential of p21 as a gene therapy treatment for prostate cancer, by introducing p21 into both androgen-dependent (AD) and independent (AI) human prostate cancer cell lines via a recombinant adenoviral vector, Ad5CMV-p21, carrying human p21 cDNA. MATERIALS AND METHODS: The LNCaP, DU145 and PC-3 human prostate cancer cell lines were cultured and infected with Ad5CMV-p21. Cell growth, cell-cycle progression and tumorigenicity were then assessed by thymidine incorporation into cellular DNA, and cell number, flow cytometry, and tumour growth after inoculating the cells into nude mice. RESULTS: Growth was inhibited in Ad5CMV-p21 viral-infected AD and AI prostate cancer cells. The effects were dose-dependent, regardless of the androgen status of the cell lines. Flow cytometric analysis showed that Ad5CMV-p21 arrested cell-cycle progression at G1/S with no appreciable effect on the levels of apoptotic cells. The tumorigenicity of cancer cells infected with Ad5CMV-p21 was greatly reduced in athymic mice. CONCLUSIONS: These results suggest that Ad5CMV-p21 may be a new therapeutic agent for human prostate cancer gene therapy. PMID- 12887491 TI - Ex-vivo comparison of the haemostatic properties of standard transurethral resection and transurethral vaporization resection of the prostate. AB - OBJECTIVE: To compare the haemostatic properties of standard transurethral resection of the prostate (TURP) and transurethral vaporization resection of the prostate (TUVRP), as perioperative bleeding is still regarded as the major complication of prostate resection. MATERIALS AND METHODS: Isolated blood perfused porcine kidneys were used to determine the haemostatic efficacy of TURP and TUVRP (using two different electrodes). Bleeding was quantified precisely in relation to tissue ablation for the two techniques, and specimens were evaluated histologically. RESULTS: Both TUVRP groups had significantly less bleeding (P = 0.005) than the TURP group for a standardized ablation volume of perfused kidney tissue (18.9, 19.5 and 24.1 mL/min, respectively). The different TUVRP electrodes had no significant haemostatic differences. The histology showed significantly (P = 0.03) larger coagulation zones for the TUVRP groups than for standard TURP. CONCLUSIONS: TUVRP ex-vivo was associated with significantly better haemostasis than TURP. The haemostatic properties of different active electrodes for TUVRP seem to be equivalent. PMID- 12887492 TI - Libido and desire: join the club? PMID- 12887493 TI - A minimally invasive technique for outpatient local anaesthetic administration of intradetrusor botulinum toxin in intractable detrusor overactivity. PMID- 12887494 TI - Safety and efficacy of 12-mm radial dilating ports for laparoscopic access. PMID- 12887495 TI - Day-case holmium laser enucleation of the prostate for gland volumes of < 60 ml: early experience. PMID- 12887496 TI - Pharmaceutical review. PMID- 12887497 TI - A novel technique for ureteric access in ileal conduits. PMID- 12887498 TI - No-scalpel vasectomy: a cautionary tale of failure. PMID- 12887499 TI - Recurrence and progression in stage T1G3 bladder tumour with intravesical bacillus Calmette-Guerin (Danish 1331 strain). PMID- 12887500 TI - Skills training and the role of skills centres. PMID- 12887504 TI - Some new aspects of the pathology, pathogenesis, and aetiology of disseminated lung lesions in slaughter pigs. AB - From 40 pigs rejected for human consumption at slaughter due to an apparent presence of pyemic lung lesions (defined as disseminated processes containing pus and/or necrotic material), the lungs, spleen, liver, and kidneys were subjected to an extended macroscopic examination. Several lung lesions were sampled from each animal for histological and bacteriological examination. Samples from the kidneys and spleens were also subjected to bacteriological examination. At gross level, four groups of lung lesions were identified: 1) disseminated foci with contents of pus and/or necrotic material (n=26); 2) disseminated or multifocally located ecchymoses with a central area of fibroplasia (n=9); 3) non-pneumonic lesions, i.e., disseminated areas of atelectasis (n=1) or haemorrhagic areas developing due to the process of slaughter (n=1); and 4) suppurative lesions without a disseminated distribution pattern (n=3). Histologically, the disseminated suppurative/necrotic foci were identified as: A) abscesses (n=10); B) necrotic lesions (n=6); and C) ectatic or ectatic-like bronchioles with contents of pus and necrotic material (n=10). The macroscopic observation of disseminated centres of fibroplasia with peripheral ecchymoses (n=9) was confirmed histopathologically. The livers of five pigs contained multiple areas of chronic interstitial fibrosis related to migration of Ascaris suum larvae ("milk spotted liver"). Such hepatic lesions were significantly (p<0.01) related to the simultaneous occurrence of disseminated pulmonary ecchymoses with a central area of fibroplasia. Generally, all lung lesions of each individual animal contained identical monocultures of bacteria following this pattern: Staphylococcus aureus (abscesses); Actinomyces hyovaginalis (necroses); S. aureus, A. hyovaginalis, and Arcanobacterium pyogenes (ectatic and ectatic-like bronchioles). Areas with fibrosis were sterile or contained bacteria considered to be a result of contamination. Apart from one kidney, from which S. aureus was cultured, all other organs were sterile. It is concluded that difficulties exist in differentiating pulmonary pyemic lesions from non-pyemic lesions at the gross level. Thus, it was not possible to distinguish between abscesses/necroses and ectatic bronchioles, the pathogenesis of the latter being uncertain. However, the chronic non-pyemic lesions related to the migration of A. suum larvae should be identified by the absence of pus/necrosis. S. aureus was predominantly isolated from abscesses, whereas, and most surprisingly, A. hyovaginalis was the dominant bacterium isolated from the pulmonary necroses. PMID- 12887505 TI - Overexpression of S100A4 is closely related to the aggressiveness of gastric cancer. AB - Elevated levels of the calcium-binding protein S100A4 cause metastasis of benign rat mammary tumor cells. To investigate whether S100A4 plays an important role in the invasion and metastasis of gastric cancers, we examined the gene mutations in the coding regions and expression patterns of the S100A4 in gastric adenocarcinoma in Korea. Moderate to strong expression of S100A4 was found in 53 (68.8%) of the 77 gastric adenocarcinomas, whilst normal gastric epithelium either failed to stain or showed weak staining. Interestingly, S100A4 expression was more frequently observed in gastric cancer patients with advanced gastric cancer (p=0.039), positive lymph node metastasis (p=0.001), and peritoneal dissemination (p=0.022). No gene mutations were found in the analyzed genomic area in 77 gastric adenocarcinomas and 15 gastric cancer cell lines. We found one single nucleotide polymorphism without an amino acid change, A99G, in two cases. These data suggest that the overexpression of S100A4 may be closely related to the aggressiveness of gastric cancer in Korea. PMID- 12887506 TI - Colonization of hospital water systems by legionellae, mycobacteria and other heterotrophic bacteria potentially hazardous to risk group patients. AB - Occurrences of legionellae and nontuberculous mycobacteria were followed in water systems of a tertiary care hospital where nosocomial infections due to the two genera had been verified. The aim was to examine whether their occurrence in the circulating hot water can be controlled by addition of a heat-shock unit in the circulation system, and by intensified cleaning of the tap and shower heads. One hot water system examined had an inbuilt heat-shock system causing a temporary increase of temperature to 80 degrees C, the other was an ordinary system (60 degrees C). The heat-shock unit decreased legionella colony counts in the circulating hot water (mean 35 cfu/l) compared to the ordinary system (mean 3.6 x 10(3) cfu/l). Mycobacteria constantly present in the incoming cold water (mean 260 cfu/l) were never isolated from the circulating hot water. Water sampled at peripheral sites such as taps and showers contained higher concentrations of legionellae, mycobacteria, and mesophilic and Gram-negative heterotrophs than the circulating waters. The shower water samples contained the highest bacterial loads. The results indicate the need to develop more efficient prevention methods than the ones presently used. Prevention of mycobacteria should also be extended to incoming cold water. PMID- 12887507 TI - Multiple co-infections (Mycoplasma, Chlamydia, human herpes virus-6) in blood of chronic fatigue syndrome patients: association with signs and symptoms. AB - Previously we and others found that a majority of chronic fatigue syndrome (CFS) patients showed evidence of systemic mycoplasmal infections, and their blood tested positive using a polymerase chain reaction assay for at least one of the four following Mycoplasma species: M. fermentans, M. hominis, M. pneumoniae or M. penetrans. Consistent with previous results, patients in the current study (n=200) showed a high prevalence (overall 52%) of mycoplasmal infections. Using forensic polymerase chain reaction we also examined whether these same patients showed evidence of infections with Chlamydia pneumoniae (overall 7.5% positive) and/or active human herpes virus-6 (HHV-6, overall 30.5% positive). Since the presence of one or more infections may predispose patients to other infections, we examined the prevalence of C. pneumoniae and HHV-6 active infections in mycoplasma-positive and -negative patients. Unexpectedly, we found that the incidence of C. pneumoniae or HHV-6 was similar in Mycoplasma-positive and negative patients, and the converse was also found in active HHV-6-positive and negative patients. Control subjects (n=100) had low rates of mycoplasmal (6%), active HHV-6 (9%) or chlamydial (1%) infections, and there were no co-infections in control subjects. Differences in bacterial and/or viral infections in CFS patients compared to control subjects were significant. Severity and incidence of patients' signs and symptoms were compared within the above groups. Although there was a tendency for patients with multiple infections to have more severe signs and symptoms (p<0.01), the only significant differences found were in the incidence and severity of certain signs and symptoms in patients with multiple co infections of any type compared to the other groups (p<0.01). There was no correlation between the type of co-infection and severity of signs and symptoms. The results indicate that a large subset of CFS patients show evidence of bacterial and/or viral infection(s), and these infections may contribute to the severity of signs and symptoms found in these patients. PMID- 12887508 TI - Determination of proliferation index in neoplasms using different Ki-67 equivalent antibodies. AB - Paraffin sections from 23 tumours were immunohistochemically stained with the following four Ki-67 equivalent antibodies: monoclonal MIB-1 (DAKO), monoclonal MM1 (Novocastra), polyclonal NCL-Ki-67p (Novocastra), and polyclonal Rah Ki-67 (DAKO). Ki-67 labelling indices were determined by counting in exactly the same area in each case. MIB-1 showed the highest labelling index in 21 of the 23 cases, and the mean MIB-1 index was approximately 30% higher than that of the other antibodies. The differences between MM1, NCL-Ki-67p and Rah Ki-67 were small and non-significant. There was a positive correlation between each of the four antibodies. As these findings may be of importance when the Ki-67 labelling index is used as a criterion for tumour grading or for clinical prognostication, this necessitate identification of the antibody used in every case. PMID- 12887509 TI - Influence of endotoxin-induced acute lung injury on pulmonary innate and adaptive immunity. AB - OBJECTIVE: The aim of our study was to analyze pulmonary innate and adaptive immune responses during endotoxemia-induced acute lung injury (ALI). EXPERIMENTAL DESIGN: Female BALB/c mice were challenged by endotoxin given intraperitoneally and followed for 24 h by unrestrained plethysmographic analysis. After this period, the mice were sacrificed by CO2 anesthesia and lung histopathology, pulmonary and serum levels of pro-inflammatory cytokines, numbers of lymphocyte subsets in blood and lung, and lung-derived macrophages (Mphi) were analyzed. MAIN RESULTS: Animals with endotoxemia demonstrated significant depression of tidal volumes indicating respiratory failure compared to control mice. Lung histopathology of endotoxin-exposed animals revealed alveolar leakage characterizing ALI. Pulmonary levels of interleukin (IL)-1beta, IL-6 and interferon (IFN)-gamma in animals with endotoxemia were significantly elevated, whereas serum levels of IL-6 only were increased. IFN-gamma was strongly expressed by lung-derived Mphi with high CD11b expression, and this subset significantly increased in the lungs after endotoxin challenge. Additionally, the numbers of lung-resident CD4+ and total T-lymphocytes were significantly reduced after challenge. CONCLUSION: These data suggest that endotoxemia-induced ALI is associated with exaggerated and sustained pulmonary innate immune responses partly mediated by activated Mphi, whereas adaptive immunity in the lungs is compromised. PMID- 12887510 TI - Multiple spindle cell lipomas of the tongue: report of a case. AB - Spindle cell lipoma (SCL) is a relatively uncommon benign lipomatous tumor characterized by a mixture of mature adipocytes and bland spindle cells on a fibromyxoid background. These lipomas usually arise as solitary nodules in the subcutaneous tissue of the posterior neck, shoulder and back in adult men. Multiple SCLs account for 0.5% to 3% of all SCLs. In the literature, there have been 15 cases of intraoral SCL, including 6 cases of SCL in the tongue. We report a case of multiple SCLs on the bilateral margins of the tongue of a 75-year-old Japanese man. The tumors were removed surgically, and they exhibited the classic histopathological features of SCL, being well-circumscribed masses composed of a mixture of adipocytes and fibroblast-like spindle cells within a fibromyxoid stroma. Immunohistochemical analysis revealed that the spindle cells were strongly positive for CD 34 and bcl-2. To our knowledge, this is the first report of a case of multiple SCLs of the tongue. PMID- 12887511 TI - Immunohistochemical analysis of Omi/HtrA2 expression in stomach cancer. AB - The serine protease Omi/HtrA2 is released from mitochondria into the cytosol after apoptosis stimuli, inducing apoptosis in a caspase-independent manner through its protease activity and in a caspase-dependent manner by neutralizing the inhibition of inhibitor of apoptosis proteins (IAPs) on caspases. Alteration of apoptosis is essential for cancer development, and cancer cell death by radiation and chemotherapy is largely dependent upon apoptosis. Thus, analysis of the expression status of Omi/HtrA2, a regulator of apoptosis, in cancer tissues is needed for an understanding of cancer development. In the current study, we analyzed the expression of Omi/HtrA2 in 60 advanced gastric adenocarcinomas by immunohistochemistry using a tissue microarray approach. Immunopositivity (defined as >/=30%) was observed for Omi/HtrA2 in 43 (72%) of the 60 cancers. By contrast, the surface mucous cells and mucous neck cells in the normal gastric mucosa showed no or weak expression of Omi/HtrA2. Taken together, these results suggest that stomach cancer cells in vivo may need Omi/HtrA2 expression for apoptosis, and that Omi/HtrA2 expression might be involved in stomach cancer development. PMID- 12887512 TI - Incidence of oesophageal necrosis in an autopsy material. AB - The incidence of oesophageal necrosis was studied in 310 consecutive autopsies of patients more than 2 years old. The oesophagus and upper part of the stomach were excised and formalin-fixed in all cases suspicious of necrosis. Suspicious areas were taken out for microscopy. The necroses presented as dark coloured or black, punctate, striped or confluent areas, most frequently located in the lower third of the organ and usually stopping abruptly at the cardia. They were distinguished from autolytic changes by the presence of a vital reaction (i.e. fibrin exudation and presence of neutrophils). Necroses were observed in 32 of the 310 patients. In nine patients the necrosis extended into or through the muscularis propria. In 13 patients there was only mucosal necrosis. In the remaining cases the necroses extended into the submucosa. In one patient the necrosis caused fatal bleeding. We believe that oesophageal necrosis is a change which is often overlooked - whether presenting at endoscopy or at autopsy - and that the condition should be taken seriously as a cause of possible bleeding, perforation or septicemia. The cause is not firmly established, but the present as well as earlier works point to acid reflux combined with hypoperfusion as important pathogenetic factors. PMID- 12887513 TI - Surgeon scientists and the need for more experiments. PMID- 12887514 TI - Laparoscopic adjustable gastric banding: a real option for a real problem. PMID- 12887515 TI - Batons and the biofuture of vascular surgery. PMID- 12887516 TI - COX-2 inhibitors and breast cancer. PMID- 12887517 TI - Pattern of equestrian injuries presenting to a Sydney teaching hospital. AB - BACKGROUND: Equestrian activities are associated with a high rate of injury. Attempts to reduce the incidence and severity of injury require accurate characterization of risk factors and pattern of injury. The present study was performed to analyse the injuries seen at an Australian centre where a large number of equestrian injuries present. METHODS: Data were collected prospectively over a 3 year period on all equestrian injuries presenting to the Prince of Wales -Hospital complex. These data were compared against and combined with retrospectively collected data over the preceding 3 years at the same centre. RESULTS: Two hundred and twenty-one injured equestrians presented (181 consented) in the prospective period of the study, and 208 presented in the retrospective period. Overall, 81% of riders were wearing a helmet at the time of their injury. Helmet use was associated with a significantly lower admission rate (27% vs 55%; P < 0.0001, from combined data). Recreational equestrians had a higher admission rate than professional equestrians, and had a significantly higher head and spine injury rate than the professional group. Rate of helmet use increased from 72% in the retrospective group to 91% in the more recent prospective group, and total admissions decreased from 43% to 14%. CONCLUSION: Significant and serious injuries are associated with equestrian activities, with the higher risk group being recreational equestrians, and riders not using a helmet. The pattern of injury favours head and spine in recreational and non-helmeted riders, and extremities for professional and helmeted riders. Helmet use is still not universal among riders, although an increase in its use may be contributing to an overall reduction in admission rate. Facial and spinal injuries still occurred in helmeted patients. PMID- 12887519 TI - Sexual function after aortic surgery. AB - BACKGROUND: Male patients attending for follow up after aortic surgery have often commented that there has been a change in their sexual function. METHODS: A survey of patients attending vascular outpatient clinic and private practice following aortic surgery was carried out by personal interview. RESULTS: Of those who were sexually active before operation, 40% thought that there had been a diminution in performance and 11% thought that there was some improvement. Similar results have been found in other centres. CONCLUSIONS: Aortic surgery carries a risk of diminution in sexual function as a consequence of the operation. It is the duty of the surgeon to warn patients of this risk when seeking informed consent. PMID- 12887521 TI - Long-term review of a breast conservation series and patterns of care over 18 years. AB - BACKGROUND: Multiple randomized trials of breast conservation compared with mastectomy in early breast cancer have validated equivalence of survival. Overwhelmingly the standard management of breast conservation includes surgical removal of the tumour, axillary dissection, postoperative breast irradiation, and adjuvant systemic therapy as appropriate. The outcomes are reviewed of 832 women with early breast cancer treated by local resection and irradiation at Royal Prince Alfred Hospital over an 18 year period, with particular emphasis on the changing patterns of practice. METHODS: Between September 1978 and May 1996, 832 women with early stage breast cancer were treated with conservative surgery and radiation therapy. The changes in patient, tumour and treatment factors were analysed over this time period. The outcomes of local recurrence and survival were recorded. Trends in patterns of these variables were evaluated by dividing the 18 years accrual period into three consecutive periods (1978-1983, n = 28; 1984-1990, n = 392; 1991-1996, n = 412). RESULTS: At a median follow up of 76 months, the 5 and 10 year actuarial local recurrence rates were 4% and 6%, respectively. Half of the local recurrences were at the primary site. Young age, extensive intraduct carcinoma, oestrogen receptor (ER) status and extranodal spread were predictive of local recurrence on multivariate analysis. The 5 and 10 year overall survival rates were 88% and 73%, respectively. Actuarial 5 year local recurrence (4%, 6%, 2%) and survival (96%, 88%, 92%) rates varied little across the three time intervals. There was an increase in median age from 46 to 56 years over the accrual period, with no change in median primary tumour size (1.5 cm). There were significant histopathological improvements in reporting margin status and ER status. Surgically, the median number of axillary lymph nodes retrieved (14) did not alter significantly. With respect to adjuvant therapies, irradiation of lymph nodes regions decreased over time. The proportion of patients receiving adjuvant hormones or chemotherapy increased significantly (18%, 35%, 54%). CONCLUSIONS: The low local recurrence rate and high survival are consistent with published literature for early breast cancer. Changes in practice during the accrual period included improvements in histopathological reporting, a reduction in irradiation of lymph node regions, and an increase in the use of systemic therapy. These changes parallel international recommendations regarding the optimal management of early breast cancer. PMID- 12887523 TI - Acquired jejuno-ileal diverticular disease: a diagnostic and management challenge. AB - BACKGROUND: Acquired jejuno-ileal diverticular disease (JID), a result of abnormalities in the smooth muscle or myenteric plexus of the small bowel, is less rare than was once believed. Approximately 1.3% of the population has JID, of whom approximately 10% present with life-threatening complications such as inflammation, perforation, bleeding, obstruction and malabsorption. Jejuno-ileal diverticular disease can be diagnostically and therapeutically challenging, and complications are often diagnosed only at laparotomy, while the best management is not agreed on in the literature. To increase the awareness of this condition and its complications, the Auckland Hospital's experience of JID was reviewed. METHODS: Retrospective review was carried out of the audit data and the discharge coding records of Auckland Public Hospital for the 5 year period leading to November 2001. RESULTS: Nine cases with a variety of presentations were found. Those cases are described and a literature review of JID is provided. CONCLUSION: Jejuno-ileal diverticular disease should be included in the differential diagnosis when dealing with surgical emergencies in the elderly presenting with features of bowel perforation, obstruction or bleeding. PMID- 12887525 TI - Comparison of the use of endoscopic and radiologic gastrostomy in a single head and neck cancer unit. AB - BACKGROUND: Head and neck cancer patients frequently require gastrostomy feeding. Different insertion techniques have been described. The aim of the present study was to compare clinical results of percutaneous endoscopic and radiological gastrostomies in patients treated in a regional head and neck cancer unit. METHODS: The records of patients who received either percutaneous endoscopic gastrostomy (PEG) or percutaneous radiological gastrostomy (PRG) between August 1997 and February 2001 were reviewed retrospectively. Documented complications (leak, infection, nausea and vomiting, ileus, bleeding, peritonitis) were recorded, compared and evaluated. RESULTS: There were 74 patients (56 PEG, 18 PRG), most with stage III and IV head and neck malignancy. There was a significantly lower incidence of complications in PEG than PRG (11% vs 44%, P = 0.004). There was a delay of feeding due to tube placement in 4% of PEG and 22% of PRG (P < 0.025). Major complications occurred in 3.6% and 5.6% of PEG and PRG, respectively. Generally the complication rate for either form of gastrostomy was comparable with other studies. No procedure-related deaths occurred. CONCLUSION: Selection bias, technique and tube type appeared to influence the complication rate in the present review. Percutaneous endoscopic gastrostomy will remain the authors' preferred method while PRG will be reserved for those cases for whom endoscopic placement is deemed to be impractical. PMID- 12887526 TI - Laparoscopic adjustable gastric banding: initial Tasmanian experience. AB - BACKGROUND: The laparoscopic adjustable gastric band (LAGB) is a minimally invasive, adjustable and reversible bariatric procedure. The present paper reports an initial 2 year experience at Royal Hobart Hospital, Tasmania. METHODS: Between February 1999 and June 2001, 207 patients underwent LAGB insertion (176 female, 31 male). The mean age was 43 years (range: 16-74 years). Mean preoperative weight was 125 kg (range: 83-210 kg) and mean body mass index (BMI) was 45.9 (range: 32.6-67.0). The Bioenterics LAGB (Inamed, Chullora, NSW, Australia) was used in all cases. The average follow up was 17 months (range: 3 24 months). Three patients were lost to follow up (1.5%). RESULTS: The average weight loss was 12.4 kg at 3 months, 25.3 kg at 1 year and 34.8 kg at 2 years. The average BMI was reduced from 45.9 preoperatively to 41.3 at 3 months, 36.9 at 1 year and 33.5 at 2 years. Reoperation for band slippage occurred in 24 patients (11%), and the injection reservoir required changes in 22 patients (11%). There were three perforations while inserting the LAGB, two non-fatal pulmonary emboli (1%) and two cases of deep vein thrombosis (1%). There has been no mortality. CONCLUSIONS: Laparoscopic adjustable gastric banding is a safe and effective method of achieving significant (P < 0.0001) weight reduction in the morbidly obese. PMID- 12887528 TI - Imprint cytology versus frozen section: intraoperative analysis of sentinel lymph nodes in breast cancer. AB - BACKGROUND: Intraoperative analysis of the sentinel node status in breast cancer adds the benefit of proceeding to axillary dissection at the same time as the primary breast operation, without the need for a second trip to theatre. However, the method of intra-operative analysis must be both rapid and accurate for use in this setting. METHODS: A prospective series of 20 consecutive patients in one hospital who underwent sentinel node biopsy was performed. Intraoperative evaluation by imprint cytology and frozen section was compared with the final histopathological results. The time taken for both procedures was recorded. RESULTS: A total of 21 sentinel node procedures was performed. The sentinel node was found in 20 procedures (95%). The average time to perform intraoperative pathological analysis was 17 min (range: 5-35 min). Of the eight positive sentinel nodes, five were positive on imprint cytology alone and five were positive on frozen section alone (62.5% accuracy). The statistical false negative rate was 18.8% and the 'false reassurance rate' (patients with an initially negative node that was subsequently positive on formal histology who required a second trip to theatre for axillary dissection) was 37.5%. All patients with a negative sentinel node had a negative axilla. CONCLUSIONS: Intraoperative analysis of sentinel lymph nodes in breast cancer can be performed quickly. Imprint cytology and frozen section show comparable accuracy in predicting sentinel node status. PMID- 12887529 TI - Alternative sites of injection for sentinel lymph node biopsy in breast cancer. AB - BACKGROUND: Sentinel node biopsy is rapidly gaining popularity as a less invasive approach to nodal staging in breast cancer. The optimal route of injection of radiocolloids and dye is controversial. The purpose of the present paper was to review and assess the literature. METHODS: A MEDLINE search for reports of studies involving different injection sites of colloid and/or dye was performed. RESULTS: Although controversial, current evidence suggests that subareolar (SA) or intradermal/subdermal (ID/SD) injection will map the same axillary sentinel nodes (SN) as peritumoral (PT) injection in the vast majority of cases, is at least as successful, and is better logistically. Peritumoral, but not alternative routes, identify extra-axillary sentinel nodes, which are important in a minority of patients. CONCLUSIONS: It is recommended that at least some of the radiocolloid be injected peritumorally to avoid missing those SN not located in the ipsilateral axilla. Injection of the dye and a portion of radiocolloid in an ID/SA location is reasonable to take advantage of the general ease and accuracy of ID/SA injections in identifying axillary SN. PMID- 12887530 TI - Long-term comparison of external valvular stenting and stripping of varicose veins. AB - BACKGROUND: The recurrence rate following stripping of the long saphenous vein is unacceptable. Repair of the terminal valve of the long saphenous vein is a physiological alternative. The aim of the present study was to prospectively compare the long-term recurrence rates in patients having had reparative and ablative procedures at the same time. METHODS: Patients were grouped first into those receiving high ligation and stripping of the long saphenous vein in one limb and valve repair on the contralateral side. The second group consisted of those patients who received short saphenous vein removal and valve repair of the ipsilateral long saphenous system. Finally, the effect of pregnancy on patients who received valve repairs was also studied. All patients were examined postoperatively clinically and with duplex ultrasound. RESULTS: Cumulative and absolute recurrence rates for both procedures were analysed. Over the 13-year follow-up period, external valvular stenting produced a superior outcome. The effect of pregnancy on the venous valve repair cases resulted in recurrences in 23% of cases but there was no evidence of incompetence at the site of the valve repair. CONCLUSION: If a valve repair is possible at the saphenofemoral junction, then it should be the preferred treatment option. PMID- 12887531 TI - Routine circumcision practice in Western Australia 1981-1999. AB - BACKGROUND: Routine neonatal circumcision has declined in most English-speaking countries. The purpose of the present study was to investigate the trends in incidence of routine circumcision in Western Australia and current patterns of the procedure according to sociodemographic factors. METHODS: Hospital morbidity data were used to conduct a population-based study of all circumcisions performed in Western -Australian hospitals during 1981-1999. Medicare insurance rebate statistics were used to estimate the numbers of circumcisions performed outside of hospital on boys <6 months of age between 1994 and 1999. RESULTS: In 1994, 9.8% of boys were circumcised before reaching 6 months of age, falling to 7.9% by 1999. Boys <6 months of age were 3.3 times more likely to undergo a hospital performed circumcision if they lived in country areas compared with metropolitan Perth. Middle socioeconomic class families were twice as likely to circumcise a son than those in the higher and lower socioeconomic groups. The decreasing circumcision rate in boys <6 months of age was partly offset by an increase in routine circumcisions in boys older than 6 months of age during 1981-1999. CONCLUSION: Circumcision remains a relatively common procedure in Western Australia. Based on total routine circumcision rates in 1999, 10.2% of boys will be circumcised by the time they reach 15 years of age. The routine circumcision rate in boys <6 months is falling in contrast to a rising routine circumcision rate in older boys. Risk factors for a circumcision before 6 months of age include living in country areas and a middle socioeconomic status. PMID- 12887532 TI - Preoperative cardiac assessment for patients with infrarenal abdominal aortic aneurysms: a survey of current practice by vascular surgeons in New South Wales and Australian Capital Territory. AB - BACKGROUND: The reported mortality rate following open elective repair of abdominal aortic aneurysm (AAA) varies between 0 and 12%. Much of the mortality and major morbidity is caused by cardiac events. The evidence regarding best practice for cardiac assessment and optimization of this patient group is unclear. The aim of the present study was to evaluate current practice of cardiac risk factor assessment by vascular surgeons in New South Wales (NSW) and Australian Capital Territory (ACT) for patients undergoing open elective repair of infrarenal AAA. METHODS: A postal questionnaire was sent to 46 surgeons in NSW and ACT identified as expressing a principal or major interest in vascular surgery. If no response was received within 3 weeks, a second questionnaire was sent and if no response was received after the second mailing, a telephone survey of non-responders was conducted. Data were collated regarding the importance of risk factors elicited by clinical history, preoperative investigation, referral for cardiological opinion, use of perioperative beta-blockade and the timing of aortic surgery in relation to coronary artery revascularization and acute myocardial infarction. RESULTS: The overall response rate was 87% (40/46) and the median (range) response time was 14 (4-109) days. Only 22 of 40 and 23 of 40 surgeons consider diabetes mellitus or renal impairment, respectively, to be important when assessing cardiac risk and 34 of 40 surgeons do not employ a validated risk index in preoperative assessment. Sixteen of 40 surgeons refer all patients needing AAA repair to a cardiologist, while 24 of 40 would initiate cardiac investigations themselves (either stress electrocardiography, scintigraphy or echocardiography). Seventeen surgeons always or usually commenced perioperative beta-blockade with wide variations in the commencement (1 to > 28 days preoperatively) and duration (< 1 week to > 28 days postoperatively) of treatment. The timing of AAA repair following coronary revascularization ranged from < 1 week to 6 months and delay in surgical repair of AAA following myocardial infarction ranged from < 1 week to > 6 months. CONCLUSION: Preoperative assessment of cardiac risk in patients for repair of AAA lacks consensus among vascular surgeons in NSW and ACT. The diversity of clinical practice may rest with the paucity of prospective trials published in the medical literature or the influence of local institutional facilities. PMID- 12887533 TI - Squamous cell carcinoma of the lip: is there a role for adjuvant radiotherapy in improving local control following incomplete or inadequate excision? AB - BACKGROUND: The treatment of squamous cell carcinoma of the lip with surgery is usually curative but incomplete/inadequate excision may be associated with recurrence and poor outcome. There is no consensus in the literature on the definition of an adequate excision margin. METHODS: Patients treated for squamous cell carcinoma of the lip at Westmead Hospital, Sydney, between 1980 and 2000 were eligible for inclusion. Polytomous logistic regression analysis was undertaken to assess for predictors of recurrence. Recurrence-free and overall survival were calculated using Kaplan-Meier survival curves. RESULTS: A total of 130 patients was identified. Median age at diagnosis was 64 years (23-97 years). Most lesions (90%) were located on the lower lip in 96 (74%) male patients. Median follow-up duration was 54 months (0-189 months). Most patients -presented with T1 lesions (75%). Initial treatment was surgery (39%), radiotherapy (48%) or both (13%). Twenty-seven per cent of excised lesions had a close (< or =2 mm) or positive margin. A total of 40 patients (31%) had recurrence (18% lymph nodes, 11% lip and 2% both). In the surgery group recurrence was significantly more likely with close or positive margins (P = 0.05). The 2 year -recurrence-free survival was 82% and 54% for radiotherapy and surgery, respectively (P < 0.001). The 2 year overall survival was similar (90% radiotherapy vs 100% surgery; P = 0.58). CONCLUSION: Incomplete or inadequate excision of some lip cancers results in local recurrence. If re-excision is not feasible -surgeons should consider the role of adjuvant radiotherapy in improving local control. PMID- 12887534 TI - Use of adhesive surgical tape with the absorbable continuous subcuticular suture. AB - BACKGROUND: The absorbable continuous subcuticular suture is frequently used to close surgical incisions where the aim is healing by primary intention. A form of adhesive surgical tape is commonly also placed over the wound but this combination closure seems to have its development based on anecdotal, rather than experimental evidence. The present study reviews the scientific literature on the development of sutureless wound closure and presents the current evidence for the use of combination wound closure. METHODS: Review was undertaken of the medical literature using the PubMed Internet database and cross-referencing major articles on the subject. The following combinations of key words were searched: skin closure, wound closure, suture technique, sutureless, adhesive tape, op site, staples, subcuticular suture, complication, infection and scars. RESULTS: Taped closure alone has advantages of lower wound infection rates and greater wound tensile strength, but disadvantages of epidermal reaction, skin edge inversion, doubtful safety and time required for meticulous surgical technique. The use of the continuous absorbable subcuticular suture allows accurate skin edge approximation, which increases the safety margin. The combination closure has a slightly superior cosmetic result to sutureless techniques but no study has been performed to compare the results of combination subcuticular suture and tape, with tape or subcuticular suture alone. CONCLUSIONS: There is no evidence in the scientific literature to justify or support the practice of closing a surgical wound with both subcuticular suture and adhesive surgical tape. PMID- 12887535 TI - Training surgeon scientists. AB - Beginning in 1995, College research resources were orientated towards the support and training of surgeon scientists. A questionnaire was sent to recipients of College awards in order to gauge a measure of outcome of this policy. Although it is early days, a significantly high number of awardees have either completed or are completing research degrees. There has been a successful allocation of National Health and Medical Research Council funding to the College surgeon scientists and a large number are already in academic positions. A significant number of publications have resulted from the College-funded research. At this early stage, the programme for creating surgeon scientists appears to be on track. PMID- 12887536 TI - Prioritizing patients for elective surgery: a systematic review. AB - BACKGROUND: Priority scoring tools are moot as means for dealing with burgeoning elective surgical waiting lists. There is ongoing development work in New Zealand, Canada and the UK. This emerging international perspective is invaluable in determining the application of these tools and addressing any pitfalls. METHODS: A systematic electronic literature review was performed. Information was also retrieved using a search of reference lists of all papers included in the review and contact with those who were involved in the development of such criteria. RESULTS: The ethical basis of prioritization differed among priority scoring tools and in a number was not stated. The majority of tools covered criteria for specific procedures. Delphi consensus methods and regression were the predominant methods for -deter-mining -specific criteria. Authors' opinions were the main source of generic criteria. Linear and non-linear models or matrices sum-mated criteria. CONCLUSION: There is debate over the ethical basis for prioritization. It is a concern that it is not addressed in many studies. The development of generic criteria showed a dearth of consensus approaches that represents a significant gap in our knowledge. On the aspects of summation and weighting, the impact of assumptions on the prioritization of patients may not have been fully explored. PMID- 12887537 TI - Endoluminal thoracic aortic repair: does the parachute need to be randomized? AB - Development of endoluminal repair has substantially reduced mortality and morbidity associated with repair of the thoracic aorta. It has significant advantages over conventional surgical repair, particularly in the management of acute dissection and thoracic aortic injury. Despite these perceived advantages it is important to realize that no level 1 data are available to prove safety and/or efficacy. Because the advantages are perceived to be so great these data are unable to be accrued and it is important that clinicians involved with this procedure keep rigorous data and undertake ongoing evaluation to ensure that clinical perceptions are eventually underpinned by robust data. PMID- 12887538 TI - Anterior rectal wall excision for endometriosis using the circular stapler. AB - BACKGROUND: Endometriosis involving the rectum is rare but is associated with significant symptoms that are best relieved by resection of the involved segment of rectum. Resection necessitates either a segmental or anterior rectal wall excision with sutured closure. Application of a circular stapling device allows an alternative technique to resect endometriosis in this area. METHOD: Following laparoscopic ablation of endometriosis elsewhere in the pelvis, the rectum must be mobilized around disease present on the anterior rectal wall. This will involve lateral and anterior extraperitoneal rectal dissection; the latter dissection mobilizing the vagina from the rectum by a sufficient length necessary to allow imbrication of the diseased area. Insertion of a circular stapler per anus allows the diseased area to be imbricated into the stapler, resulting in simultaneous excision and closure of the anterior rectal wall. RESULTS: Thirty patients with anterior rectal wall endometriosis, estimated at <2 cm in diameter and not involving > one-third of the total circumference of the rectum, have undergone successful management using this technique. Morbidity occurred in four patients, with one patient requiring further surgery. CONCLUSIONS: Laparoscopic disc excision of deposits of endometriosis involving the anterior rectal wall can be safely performed utilizing the circular stapler without the need for open surgery, and with low morbidity. PMID- 12887539 TI - Simple technique for identifying the linea alba in lower abdominal incisions. AB - BACKGROUND: Identification of the true midline in infra-umbilical longitudinal incisions is often difficult. Traditional methods of identification can be unreliable. METHODS: An alternative technique for identifying the linea alba, based on the attachments of the median umbilical ligament, is presented. RESULTS: The technique is both reliable and reproducible in identifying the midline. CONCLUSION: This technique is recommended as a means of avoiding muscle incision and facilitating wound closure. PMID- 12887540 TI - Second Sir George Montario Bedbrook Oration-1999. Some milestones in the life of George Bedbrook. Their relationship to management and research of spinal cord injuries. PMID- 12887541 TI - Is improving local control sufficient indication for radiotherapy in the management of rectal cancer? A response. PMID- 12887543 TI - Initial experience with cultured epithelial autografts in massively burnt patients. PMID- 12887544 TI - Pseudomembranous colitis requiring surgical intervention following triple therapy for Helicobacter pylori eradication--a cautionary note. PMID- 12887545 TI - Colonoscopic surveillance after surgery for colorectal cancer. PMID- 12887546 TI - Bilateral subtotal laparoscopic adrenalectomy for phaeochromocytoma. PMID- 12887547 TI - Detection of prostate cancer by magnetic resonance imaging and spectroscopy in vivo. PMID- 12887548 TI - Sigmoid colon perforation following a migrated biliary stent. PMID- 12887551 TI - A measure for the assessment of eating disorders: reliability and validity studies of the Rating of Anorexia and Bulimia interview - revised version (RAB R). AB - The Rating of Anorexia and Bulimia interview (RAB) is a Swedish semi-structured interview for clinical and research purposes for a wide range of eating disorder symptoms and related psychopathology. The objectives were to evaluate the reliability and validity of a revised RAB version (RAB-R). The RAB-R was assessed in terms of internal consistency, inter-rater and test-retest reliability, and criterion and convergent validity. Samples included a clinical sample of eating disorder patients (n = 71) and a sample of randomly drawn female controls (n = 31). The RAB-R showed satisfactory internal consistency, inter-rater and test retest reliability, correlated well with related measures, and discriminated between patients and normal controls. We conclude that the RAB-R is a promising interview instrument and continued evaluation should focus on comparing subgroups of eating disorder patients with other clinical groups. PMID- 12887552 TI - The relationship between parental physical availability and child sexual, physical and emotional abuse: a study among a sample of university students in South Africa. AB - The relationship between perceived parental physical availability and child sexual, physical and emotional abuse was investigated. The sample comprised 722 undergraduate students of psychology at the University of the North, South Africa. Participants filled in a retrospective self-rated questionnaire in a classroom setting. The questionnaire asked about perceived parental physical availability during childhood, and childhood sexual, physical and emotional abuse. Logistic regression analysis showed that, among all the participants, the factors "the participant not living with the natural (biological) mother until he or she was at least 16 years old" and "have ever had a stepfather or adoptive father until he or she was at least 16 years old" predicted child sexual abuse; and "have ever had a stepfather or adoptive father until he or she was at least 16 years old" predicted child emotional abuse. None of the other aspects of parental availability considered predicted child physical abuse. Mental health and social workers, educators and law enforcement agencies dealing with prevention and protection against child abuse should take note of the above identified predictors while designing programs for the eradication of child sexual, physical and emotional abuse. PMID- 12887553 TI - From clinical towards research interview: parenting problems with troubled adolescents. AB - The aim of this study was to investigate whether clusters of common questions asked in clinical parenting interviews with parents of troubled adolescents could contribute to the empirical assessment of parenting dimensions. Questions were derived from clinical experience and theory, and were presented in an interview with 101 parents of adolescents referred to child psychiatric outpatient clinics for emotional and behavioral disorders. Fourteen questions were formed within the parenting domain of warmth, 12 questions from the domain of control, five questions were about autonomy granting and six questions were about commitment. Scores clustered into nine subgroups: Contact, Empathy, Appreciation, Setting Limits, Consistency, Monitoring, Parenting Priority, Involvement and Autonomy Granting. Raters also made an overall judgment of the presence of parental negative attributions during the interview. The inter-rater reliability, internal consistency, and convergent validity of the subgroups were satisfactory, and the bias of socially desirable responding was limited. Directions for further research are suggested. PMID- 12887554 TI - General self-efficacy, dental anxiety and multiple fears among 20-year-olds in Norway. AB - This paper examines the extent to which low general self-efficacy and painful dental and medical experiences are related to dental anxiety, multiple fears and to avoidance of dental care. A total of 754 20-year-olds completed a series of questionnaires, including the General Self-efficacy Scale (GSE), Geer Fear Scale (GFS) and Dental Fear Survey (DFS). Females had lower self-efficacy, higher dental anxiety and higher scores on the GFS than males. Multivariate analyses (linear stepwise regression) indicated that painful dental experiences, a high score on the GFS and negative opinions about own dental health explained 37% of the variance in DFS scores. Self-efficacy had no predictive power for dental anxiety, and only dental anxiety had predictive power for dental avoidance behavior. Thirty-eight percent of the total variance in GFS scores was explained by the following variables: being a female, high dental anxiety (DFS), low general self-efficacy (GSE) and low educational level of the mother. PMID- 12887555 TI - Memories of mother, complementarity and shame: predicting response to subliminal stimulation with "Mommy and I are one". AB - A poorly understood finding with potentially wide-ranging implications is that subliminal stimulation with "Mommy and I are one" affects behavior. In this study (n= 62), "Mommy and I are one" lowered implicit mood (p= 0.0015) in comparison with a neutral stimulus ("People are walking"). The effect was most pronounced in shame-prone participants with less positive memories of their mother, and low self-mother complementarity (interaction p= 0.0044). Effects of a potentially shame-inducing stimulus ("I am completely isolated") were not significant (ps > 0.11). The results concerning less positive memories of mother replicate previous findings. We suggest that activation of unconscious associative networks explains the data. Though more research is needed, the cognitive content of these networks may involve representations of self-with-mother; for some, the affective content could involve shame. PMID- 12887556 TI - Individual differences in children's emotion understanding: effects of age and language. AB - Over the last two decades, it has been established that children's emotion understanding changes as they develop. Recent studies have also begun to address individual differences in children's emotion understanding. The first goal of this study was to examine the development of these individual differences across a wide age range with a test assessing nine different components of emotion understanding. The second goal was to examine the relation between language ability and individual differences in emotion understanding. Eighty children ranging in age from 4 to 11 years were tested. Children displayed a clear improvement with age in both their emotion understanding and language ability. In each age group, there were clear individual differences in emotion understanding and language ability. Age and language ability together explained 72% of emotion understanding variance; 20% of this variance was explained by age alone and 27% by language ability alone. The results are discussed in terms of their theoretical and practical implications. PMID- 12887557 TI - Primed word-fragment completion and successive memory test performance in normal aging. AB - Young and old subjects were investigated to examine whether: the effects of priming are influenced by aging; there is independence between primed word fragment completion and recognition performances; and the dependence between different tests is influenced by aging. A successive test paradigm was employed involving repeated assessment of to-be-remembered words by means of recognition and primed word-fragment completion. The results show that implicit memory declines with increasing age, and that correlations between different memory tests decrease with age. The outcome suggests that age-related memory decline involves several forms of memory, including primed word-fragment completion, and is reflected in correlations between measures of implicit and explicit memory. PMID- 12887558 TI - Similar effects of attention directed to acoustic and tactile stimuli on prepulse inhibition of acoustic startle. AB - Prepulse inhibition (PPI) is assumed to index automatic and controlled processing. In three experiments (n= 32, 22, and 30) participants were asked to judge the duration of a prepulse in comparison with a stimulus presented 4000 ms before the prepulse. A distracter was presented simultaneously with the prepulse to increase the cognitive demands of the task. PPI was assessed at stimulus onset asynchronies (SOAs) of 30-150 ms, and 420 ms. The prepulse was either a tone (60 dB) or a tactile stimulus (21 kPa), and startle was elicited by 95 dB white noise. Directing attention to the prepulse increased PPI at SOAs of 60 ms and longer in all experiments, but the sensory modality to which attention was directed played only a minor role. We conclude that directing attention to both acoustic and tactile prepulses increased PPI. PMID- 12887559 TI - Questions of culture, age and gender in the epidemiology of suicide. AB - Cultural values were examined as predictors of suicide incidence rates compiled for men and women in six age groups for 33 nations for the years 1965, 1970, 1975, 1980, and 1985. Hofstede's cultural values of Power-Distance, Uncertainty Avoidance, and Masculinity (i.e., social indifference) were negative correlates of reported suicide, and Individualism was a strong positive correlate. The proportion of variance in suicide reports generally related to these four cultural values was R2 = 0.25. Suicide by women and by middle-aged people was most related to cultural values, even though international variance in suicide is greater for men and for the elderly. Suicide incidence for girls and young women showed unique negative correlations with Individualism. For all age groups, Individualism predicted a greater preponderance of male suicides, and Power Distance predicted more similar male and female suicide rates. Social alienation and Gilligan's feminist theory of moral judgment were hypothesized to explain some gender differences. PMID- 12887560 TI - Parents' adjustment to late abortion, stillbirth or infant death: the role of causal attributions. AB - The aim of the present study was to explore the attributional processes and the effect of various causal attributions on post-traumatic symptomatology among 110 parents who had lost an infant. The attributional processes were examined with longitudinal data collected over 12 months. One to four weeks post-loss, approximately half of the parents reported that they were never or rarely concerned with attributing responsibility for their baby's death. The importance of attributing responsibility was associated with several post-traumatic symptoms, as was searching for meaning in the death. Attributions to oneself, others, or God were positively and significantly associated with numerous post traumatic symptoms. The results are discussed with reference to other studies dealing with the effect of attributions on subsequent adjustment among victims of trauma. PMID- 12887561 TI - IQ stability: the relation between child and young adult intelligence test scores in low-birthweight samples. AB - The stability of IQ from childhood to adulthood in low-birthweight subjects was measured in two independent samples with follow-up intervals of approximately 14 and 9.5 years. In both samples, intelligence was assessed with the WISC at a mean age of 9.5. Twenty-six subjects were retested with the WAIS at a mean age of 23.5, and 78 subjects with the BPP (the Danish Military Draft Board Intelligence Test) at the age of 19.1. Both samples obtained childhood and adult test scores below the expected means. For the Wechsler Verbal, Performance and Full-Scale IQs, the stability quotients were 0.86, 0.86, and 0.89 in the WAIS sample, and the retest correlations for the three IQs with the BPP score were 0.66, 0.65, and 0.74. Thus, the majority of children showed stable patterns of intellectual development from middle childhood to young adulthood. PMID- 12887563 TI - Characteristics of ovarian follicle development in domestic animals. AB - In most domestic animals the later stages of follicle development occurs in a wave-like pattern during oestrous cycles (cattle, sheep, goats, horses and buffalo) or periods of reproductive activity (llamas and camels). A follicle wave is the organized development of a cohort of gonadotrophin-dependent follicles all of which initially increase in size, but most of which subsequently regress and die by atresia (subordinate follicles). The number of remaining (dominant) follicles is specific to the species and is indicative of litter size. Follicle waves develop during both luteal and follicular phases and it is the dominant follicle(s) of the last follicular wave that ovulates. However, there are cases where dominant follicles from the last two follicle waves can ovulate (sheep and goats). There are exceptions to the organized wave-like pattern of follicle growth where follicle development is apparently continuous (pigs and chickens). In these animals many follicles develop to intermediate diameters and at specific times follicles that are destined to ovulate are selected from this pool and continue growing to ovulation. Understanding the pattern of follicle development in different species is increasingly important for designing improved methods to manipulate reproduction in domestic animals. PMID- 12887564 TI - The insulin-like growth factor system: a key determinant role in the growth and selection of ovarian follicles? a comparative species study. AB - The aim of the present paper is to make a comparative study of the expression of the elements of the insulin-like growth factor (IGF) system in different mammalian species and thus illuminate their potential role in the process of ovarian folliculogenesis in mammals. In most mammalian species, IGFs and IGFBPs (in particular IGFBP-2 and IGFBP-4) are considered, respectively, as stimulators and inhibitors of follicular growth and maturation. In mammalian species, IGFs might play a key role in sensitizing ovarian granulosa cells to FSH action during terminal follicular growth. Concentrations of IGFBP-2 and IGFBP-4 in follicular fluid strongly decrease and increase during follicular growth and atresia, respectively, leading to an increase and a decrease in IGF bioavailability, respectively. The decrease in these IGFBPs is because of a decrease in mRNA expression (IGFBP-2) and an increase in proteolytic degradation by PAPP-A in follicular fluid (IGFBP-2, IGFBP-4 and IGFBP-5), and likely participates in the selection of dominant follicles. In contrast, levels and/or sites of expression of IGF-I, IGF-II, IGFBP-4, IGFBP-5 and type II receptor in follicular cells strongly differ between mammalian species, suggesting that these phenomena might play species-specific or secondary roles in ovarian folliculogenesis. PMID- 12887565 TI - Oocyte and embryo quality: effect of origin, culture conditions and gene expression patterns. AB - In general, the majority of immature bovine oocytes fail to develop to the blastocyst stage following maturation, fertilization and culture in vitro. The evidence suggests that while culture conditions during in vitro embryo production can impact on the developmental potential of the early embryo, the intrinsic quality of the oocyte is the key factor determining the proportion of oocytes developing to the blastocyst stage. In addition, evidence suggests that the period of post-fertilization embryo culture is the most critical in determining blastocyst quality. This paper reviews the current literature, with emphasis on the bovine model, demonstrating evidence for an effect of oocyte origin and/or in vitro maturation conditions on the developmental capacity and gene expression patterns in the oocyte. Furthermore, the well-documented effects of post fertilization culture environment on embryo gene expression and quality are highlighted. PMID- 12887566 TI - Manipulation of domestic animal embryos and implications for development. AB - Assisted reproductive technologies, as applied to domestic animals, can exert both novel and wide-ranging influences on the development, viability and welfare of offspring. Some of the changes are evident immediately or soon after the time at which a manipulative procedure is carried out, while other changes may not be evident until later in development or, perhaps, may remain undetected throughout an animal's lifetime. The present review explores some of the consequences - in terms of foetal, placental, neonatal and post-natal effects - of exposing embryos of cattle, sheep and other species to in vitro culture per se or, during culture, to physically invasive technologies including gene injection and nuclear transfer. The innate sensitivity of oocytes and recently fertilized eggs to their in vitro environment is illustrated by an examination of the later developmental repercussions resulting from apparently innocuous choices related to in vitro culture medium formulations. In contrast, an inherent resilience and paradoxical readiness to resume development following the traumas of nuclear transfer procedures is also in evidence. The extent to which assisted reproductive technologies will succeed, where relevant, in the domestic animal sector will be influenced by our appreciation of embryo requirements, for both short- and long term developmental fitness, during their earliest developmental stages. Evidence of species-specific needs is testimony to the challenges ahead. Ultimately, our ability and inclination to resolve the limitations associated with current procedures will probably be greatly enhanced if predictive indicators (genetic, epigenetic or functional markers) of later developmental fitness can be identified. PMID- 12887567 TI - Embryo-maternal communication in bovine - strategies for deciphering a complex cross-talk. AB - Early embryonic development, implantation and maintenance of a pregnancy are critically dependent on an intact embryo-maternal communication. So far, only few signals involved in this dialogue have been identified. In bovine and other ruminants, interferon tau is the predominant embryonic pregnancy recognition signal, exhibiting antiluteolytic activity. However, this is just one aspect of the complex process of embryo-maternal signalling, and a number of other systems are more likely to be involved. To gain a more comprehensive understanding of these important mechanisms, integrated projects involving specialists in embryology, reproductive biotechnology and functional genome research are necessary to perform a systematic analysis of interactions between pre implantation stage embryos and oviduct or uterine epithelial cells, respectively. State-of-the-art transcriptomic and proteomic technologies will identify reciprocal signals between embryos and their maternal environment and the respective downstream reaction cascades. For in vivo studies, the use of monozygotic twins as recipient animals provides elegant model systems, thus eliminating genetic variability as a cause of differential gene expression. In addition, suitable systems for the co-culture of oviduct epithelial or endometrium cells with the respective embryonic stages need to be established for functional validation of candidate genes potentially involved in the dialogue between embryos and their maternal environment. The knowledge of these mechanisms should help to increase the pregnancy rate following embryo transfer and to avoid embryonic losses. Candidate genes involved in embryo-maternal communication will also be used to define new quality criteria for the selection of embryos for transfer to recipients. Another application is the supplementation of embryotrophic factors or components of embryo-maternal signalling in optimized formulations, such as bioartificial matrices. As a long-term goal, signalling mechanisms identified in bovine will also be functionally evaluated in other species, including the human. PMID- 12887568 TI - The mitochondrial genome in embryo technologies. AB - The mammalian mitochondrial genome encodes for 37 genes which are involved in a broad range of cellular functions. The mitochondrial DNA (mtDNA) molecule is commonly assumed to be inherited through oocyte cytoplasm in a clonal manner, and apparently species-specific mechanisms have evolved to eliminate the contribution of sperm mitochondria after natural fertilization. However, recent evidence for paternal mtDNA inheritance in embryos and offspring questions the general validity of this model, particularly in the context of assisted reproduction and embryo biotechnology. In addition to normal mt DNA haplotype variation, oocytes and spermatozoa show remarkable differences in mtDNA content and may be affected by inherited or acquired mtDNA aberrations. All these parameters have been correlated with gamete quality and reproductive success rates. Nuclear transfer (NT) technology provides experimental models for studying interactions between nuclear and mitochondrial genomes. Recent studies demonstrated (i) a significant effect of mtDNA haplotype or other maternal cytoplasmic factors on the efficiency of NT; (ii) phenotypic differences between transmitochondrial clones pointing to functionally relevant nuclear-cytoplasmic interactions; and (iii) neutral or non neutral selection of mtDNA haplotypes in heteroplasmic conditions. Mitochondria form a dynamic reticulum, enabling complementation of mitochondrial components and possibly mixing of different mtDNA populations in heteroplasmic individuals. Future directions of research on mtDNA in the context of reproductive biotechnology range from the elimination of adverse effects of artificial heteroplasmy, e.g. created by ooplasm transfer, to engineering of optimized constellations of nuclear and cytoplasmic genes for the production of superior livestock. PMID- 12887569 TI - The predictive value of semen analysis in the evaluation of stallion fertility. AB - Pregnancy rates in managed horse populations depend on the innate fertility of the mares and stallions involved and on the quality of breeding management. Of course, because a single stallion usually mates many mares, stallion fertility is a critical factor in the overall success of a breeding program. Unfortunately, accurate evaluation of stallion fertility per se requires a large number of normal mares to be mated and is necessarily retrospective. Rather, the ideal is to predict fertility in advance of the stallion's breeding career, and this is currently attempted by way of a thorough physical examination and a routine analysis of semen quality. However, while such a 'breeding soundness examination' identifies stallions that clearly lack the capacity for adequate fertility, it is of limited use for predicting the level of fertility and fails to identify some seriously sub-fertile animals. Similarly, while various sperm function tests (e.g., sperm head morphometry, the hypoosmotic swelling test, glass wool-sephadex filtration, progesterone receptor exposure) have been shown to correlate fairly well with fertility in the field, most examine only a single or a narrow range of the attributes that a sperm must possess if it is to fertilize an oocyte in vivo, and are thus more useful for identifying specific causes of sub-fertility than for predicting the level of fertility. On the other hand, combining the results of the various sperm function tests does improve the reliability of fertility estimation and current research is therefore concentrated on identifying a range of tests that covers as many important sperm attributes as possible but that can be performed rapidly and cheaply. In this respect, flow-cytometry has proven to be an ideal tool because it allows the objective, rapid and simultaneous analysis of a number of properties in a large number of sperm. Moreover, stains are available for an increasing range of sperm characteristics including viability, capacitation and acrosome status, mitochondrial activity and chromatin integrity. Flow-cytometric analysis of sperm with appropriate probes thus offers considerable promise for the prediction of stallion fertility. PMID- 12887570 TI - Laboratory semen assessment and prediction of fertility: still utopia? AB - Finding a laboratory test reliable enough to predict the potential fertility of a given semen sample or a given sire for artificial insemination (AI) is still considered utopian, as indicated by the modest correlations seen between results obtained in vitro and field fertility. Male fertility is complex, and depends upon a heterogeneous population of spermatozoa interacting at various levels of the female genital tract, the vestments of the oocyte, and the oocyte itself. For this reason, laboratory assessment of semen must include the testing of most sperm attributes relevant for fertilization and embryo development, not only in individual spermatozoa but within a large sperm population as well. Strategies for the discovery of in vitro predictors of semen fertility require evaluations of low sperm doses for AI, so that differences in innate in vivo fertility can be accurately detected. PMID- 12887572 TI - What do journals do? PMID- 12887573 TI - Consistency of diet and its effects on mandibular morphogenesis in the young rat. AB - The purpose of this study was to evaluate the relationship between mandibular morphogenesis and masticatory function in 30 male Sprague-Dawley rats, 28 days of age. Animals were randomly divided into three groups and they received diets of different consistency: liquid, normal (as control group) and elastic diet. The experiment lasted 28 days, at the end of which lateral radiographs of the cranium and photographs of the hemimandibles were made. These images were magnified fourfold and analysed in two dimensions (V = vertical dimension; S = sagittal dimension). Condylar morphology was also assessed. The photographic analysis of the hemimandible showed a significantly higher growth of the mandibular ramus stimulated by the elastic diet: V2 2.4 mm, V3 2.4 mm, V4 1.9 mm, V5 4.1 mm (p < 0.05). In contrast, a statistically significant (p < 0.05) lengthening of the mandibular corpus occurred in the rats subjected to liquid diet (S1 4.5 mm, S2 2.7 mm, S3 1.6 mm, S4 2.6 mm). Measurements were taken on both the radiographic and the photographic records. The variation coefficient showed that photographic analysis was more accurate than the radiographic analysis. From these data it appears that an increase in the tonus of the protrusive muscles could be beneficial in mandibular hypoplasia and the increased tonus of the masticatory muscles in skeletal open bite. PMID- 12887574 TI - Clinical comparison between a modified light-curing denture base resin and a conventional composite resin for orthodontic bonding. AB - The purpose of this in vivo study was to evaluate and compare the bond failure rate of a modified visible light-cured denture base resin (Triad VLC Provisional Material; Dentsply International Inc., York, PA) with that of a conventional visible light-cured composite resin (Transbond XT; 3M/Unitek, Monrovia, CA) for the bonding of orthodontic brackets. Both adhesives were used in each patient following a split-mouth design. Thirty-five consecutive patients with fixed appliances were included in the study, and the performance of 655 stainless steel brackets was evaluated: 325 brackets were bonded with the modified Triad VLC resin and 330 were bonded with Transbond XT resin. The incidence and site of bond failures were recorded over a period of 12 months. The overall failure rate of Triad VLC (4.3%) was not significantly different (p>0.05) from that of Transbond XT (3.6%). No significant differences in the failure rates of upper and lower arches within each material or between the two materials were found (p>0.05). Transbond XT showed a significantly higher failure rate (p<0.05) in the anterior (4.8%) than in the posterior teeth (1.6%). The present findings demonstrate that Triad VLC could be used as an alternative bonding material for direct bonding of orthodontic brackets. PMID- 12887575 TI - Deciduous dentition-anchored rapid maxillary expansion in crossbite and non crossbite mixed dentition patients: reaction of the permanent first molar. AB - This preliminary report describes our evaluation of the behavior of permanent maxillary first molars subsequent to rapid maxillary expansion (RME) in the mixed dentition in crossbite and non-crossbite cases; maxillary deciduous canines and second molars were used as anchorage. The RME appliance was cemented on the maxillary deciduous second molars and canines in 19 (13 females, six males) patients with unilateral or bilateral permanent molar crossbites (Group A) and in 13 (10 females, three males) patients who had no crossbites (Group B). After a mean appliance activation period of 15 days for 4-8 mm expansion, the appliance was stabilized and kept in place as retention for 5-10 months. Neither active nor passive retention was applied on first permanent molars. Impressions of dental arches were made before insertion of the appliance (T1), at stabilization (T2), and at debanding (T3). Dental measurements were registered at T1, T2, and T3. In Group A, the mean maxillary permanent intermolar width was 40.6 mm at T1, 46.0 mm at T2, and 46.3 mm at T3. In Group B, 42.4 mm at T1, 46.3 mm at T2, and 45.4 mm at T3. In all subjects, the intermolar width increased during the active phase, and in Group A the crossbite was corrected and the correction was spontaneously stable. In these patients, deciduous teeth could support the heavy forces produced by the RME appliance. The maxillary intermolar distance was spontaneously increased and then maintained with no retention in Group A. The mean spontaneous expansion of permanent maxillary molars was approximately half of the mean expansion of the screw and the anchor deciduous teeth. PMID- 12887576 TI - Control of tooth movements with the Speed system. AB - Steps taken to evolve the straight-wire system eventually led to the development of self-ligating brackets. The Speed system, comprehensive of bracket, wires, force, and anchorage management, is a significant evolutionary development. The design property of the fourth wall in the Speed bracket allows the Ni-Ti wire to be placed in the slot fully. Closure of the gate activates the wire for the desired type and direction of tooth movement. A sequence of round, square, rectangular wires, Ni-Ti and stainless steel, permits a complete control of the tooth position during the different phases of the orthodontic treatment. In this study, the effectiveness of the system is investigated in four groups of patients: Class II, div 1 with four first bicuspid extractions; Class II, div 2 non-extraction; Class II div 1 non-extraction in mixed dentition; Skeletal Class III skeletal treated with an orthodontic-surgical treatment plan. Treatment results were assessed using the intra-oral photographs, and radiographs taken at the start of treatment, at the end of active treatment, and in retention. Satisfactory control of tooth positions during the horizontal, mesio-distal and torque movements, both in the extraction and non-extraction cases were observed. During the course of treatment, in extraction cases where sliding mechanics are used, shifting of the anchorage from posterior to the anterior unit is possible as a result of increased forces. This shift is facilitated by the activation of the spring clip on the anterior teeth. The advantages of the self-ligating Speed system are the low levels of force used, low friction, and precise control of tooth movements. PMID- 12887577 TI - Effect of pulse vs continuous micro-xenon irradiation on the shear bond strength of a light-cured orthodontic composite resin. AB - The aim of the study was to evaluate the effect of pulse vs. continuous irradiation on the shear bond strength of a conventional orthodontic composite resin cured with a micro-xenon light (Aurys, Degre K, Schiltigheim, France). Ninety freshly extracted bovine permanent mandibular incisors were randomly assigned to one of six groups; each group consisting of 15 specimens. Three groups were exposed to continuous irradiation of micro-xenon light for 10, 5, and 2 s, respectively, and used as controls. The remaining three groups were exposed to pulse irradiation of the same light source for 10, 5, and 2 s, respectively. After 24 h, all samples were tested in a shear mode on an Instron Machine (Instron Corp., Canton, MA, USA). Analysis was made by ANOVA with Scheffe's test for comparisons. The chi-square test was used to determine significant differences in the Adhesive Remnant Index scores. The mean shear bond strength of the brackets continuously cured for 10 s was not statistically different from that of the brackets pulse-activated for the same curing time. Also, no statistically significant differences were found between both groups cured for 5 s. The group pulse-activated for 5 s, however, had a significantly lower mean shear bond strength than the control group cured for 10 s. Finally, the group pulse-activated for 2 s showed significantly lower bond strength values than all the other groups tested. Compared with continuous light curing, the micro-xenon pulse activation provides similar shear bond strength values, except when used for only 2 s; but despite lower performance characteristics, the shear bond strength may be clinically acceptable. PMID- 12887578 TI - Prevalence of tooth-size discrepancy among different malocclusion groups. AB - This study was designed to estimate the prevalence of tooth-size discrepancy as a factor of skeletal malocclusion in the orthodontic patient population of the Campania region. The study employed the pretreatment models of 94 patients. The mesiodistal diameters of teeth were measured by digital electronic calipers (accurate to 0.01 mm) and Bolton's indices were calculated. The sample was grouped into three malocclusion groups based on the values of Steiner cephalometric analysis. Multiple regression analysis data indicated a significant linear relationship (r2 = 0.99, P = 0.0000) for Bolton's posterior, anterior and total indices. The discriminant multivariate analysis, based on stepwise Wilke's lambda, revealed five variables capable of classifying 88.6% of the sample in the four malocclusion groups. As no relationship between the four groups identified by the discriminant analysis and the types of skeletal malocclusions exists, it was unequivocally concluded that there is no evidence of any predisposition for a tooth-size discrepancy in any of the malocclusion groups. PMID- 12887579 TI - Relation between vertical facial morphology and jaw muscle activity in healthy young men. AB - The aim of the current investigation was to quantitatively analyze the relation between the activity of masticatory muscles and the inclination of the mandibular plane in a group of 73 healthy white men aged 20-36 years. The three-dimensional coordinates of soft-tissue landmarks gnathion and left and right gonion were digitized using an electromagnetic computerized instrument, the orientation of mandibular plane relative to the true vertical was computed and projected on the anatomical sagittal plane. The electromyographic (EMG) potentials of left and right masseter and temporalis anterior during maximum voluntary teeth clenching were recorded, and the mean EMG amplitude calculated. Two groups of men with opposite facial morphology were then selected: all men with a steep mandibular plane (higher than the mean plus one standard deviation) entered a first group (10 'long face' subjects), while all men with a relatively more horizontal mandibular plane (lower than the mean minus one standard deviation) entered a second group (13 'short face' subjects). Mean EMG potentials computed in the two groups were compared by using Student's t -test for independent samples. All the EMG potentials recorded during maximum voluntary clench in the 'long face' men were lower than that recorded in the 'short face' men, with statistically significant differences for all four analyzed muscles (p < 0.05). In conclusion, a non-invasive three-dimensional method confirmed that facial morphology and muscular function are significantly related, at least in men with a sound stomatognathic apparatus. PMID- 12887580 TI - Acceleration of Aux/IAA proteolysis is specific for auxin and independent of AXR1. AB - Aux/IAA proteins are short-lived transcriptional regulators involved in auxin signaling. Using Aux/IAA luciferase (LUC) fusion proteins expressed in Arabidopsis thaliana, we previously showed that rapid degradation of these proteins requires conserved Aux/IAA domain II and that exogenous auxin accelerates their degradation. To further examine auxin-mediated increases in proteolysis, the degradation of two other LUC fusion proteins, a non-cleavable ubiquitin LUC fusion (UB1-72::LUC) and SAUR15::LUC was determined in vivo in seedlings. Their half-lives were 20 +/- 4 and 104 +/- 10 min, respectively. SAUR15::LUC half-life was not affected by pre-incubation with 2,4-D. Auxin did not have an equivalent effect on UB(1-72)::LUC steady-state levels as compared to PsIAA6:LUC. LUC fused to an Aux/IAA domain II degraded more rapidly following auxin application, demonstrating that this region is sufficient for auxin mediated acceleration of proteolysis. Hormonal cross-talk at the level of Aux/IAA proteolysis was examined. 1-aminocyclopropane-1-carboxylic acid (ACC), benzyladenine (BA), abscisic acid (ABA), and brassinolide (BL) did not affect the degradation rate of IAA1::LUC, and gibberellic acid (GA3) and salicylic acid (SA) did not specifically affect the steady-state levels of Aux/IAA::LUC proteins. An Aux/IAA::LUC transgene was crossed into the auxin resistant-1 (axr1-12) background. In axr1-12, the half-life of PsIAA6(1-73)::LUC increased 4.5-fold, but proteolysis still accelerated in response to exogenous auxin. These data suggest that auxin is the only phytohormone that accelerates Aux/IAA proteolysis, and that this acceleration is specific for Aux/IAA proteins. In addition, AXR1 plays an important role in rapid basal proteolysis of Aux/IAA proteins, but is not required for auxin-mediated acceleration of their degradation. PMID- 12887581 TI - The soybean NRAMP homologue, GmDMT1, is a symbiotic divalent metal transporter capable of ferrous iron transport. AB - Iron is an important nutrient in N2-fixing legume root nodules. Iron supplied to the nodule is used by the plant for the synthesis of leghemoglobin, while in the bacteroid fraction, it is used as an essential cofactor for the bacterial N2 fixing enzyme, nitrogenase, and iron-containing proteins of the electron transport chain. The supply of iron to the bacteroids requires initial transport across the plant-derived peribacteroid membrane, which physically separates bacteroids from the infected plant cell cytosol. In this study, we have identified Glycine max divalent metal transporter 1 (GmDmt1), a soybean homologue of the NRAMP/Dmt1 family of divalent metal ion transporters. GmDmt1 shows enhanced expression in soybean root nodules and is most highly expressed at the onset of nitrogen fixation in developing nodules. Antibodies raised against a partial fragment of GmDmt1 confirmed its presence on the peribacteroid membrane (PBM) of soybean root nodules. GmDmt1 was able to both rescue growth and enhance 55Fe(II) uptake in the ferrous iron transport deficient yeast strain (fet3fet4). The results indicate that GmDmt1 is a nodule-enhanced transporter capable of ferrous iron transport across the PBM of soybean root nodules. Its role in nodule iron homeostasis to support bacterial nitrogen fixation is discussed. PMID- 12887582 TI - Rapid migration in gel filtration of the Cf-4 and Cf-9 resistance proteins is an intrinsic property of Cf proteins and not because of their association with high molecular-weight proteins. AB - Gel filtration is frequently used to study the behaviour and composition of protein complexes. In previous studies, gel filtration analysis of solubilised membranes containing the tomato Cf-4 and Cf-9 resistance proteins indicated that these Cf proteins are present in an approximately 400- and 420-kDa protein complex, respectively, which contains only one Cf molecule per complex, does not contain Rho-related proteins, and does not alter in size upon elicitation. Here, we show that inactive Cf-4 and Cf-9 mutant proteins have a similar large apparent size upon gel filtration analysis. The size remains unaltered after pre-treating the samples under harsh conditions, such as boiling with SDS and incubation in 6 m urea. A similar large apparent size was found for Cf-4 and Cf-9 isolated from SDS gel and for Cf-9 expressed by insect cells. Therefore, the large apparent size observed in our studies appears to be an intrinsic property of the Cf proteins, rather than being caused by association with high-molecular-weight protein(s). Taken together, these results suggest that caution should be taken when interpreting data obtained from gel filtration of LRR-containing proteins. PMID- 12887583 TI - The Arabidopsis mutant dct is deficient in the plastidic glutamate/malate translocator DiT2. AB - The Arabidopsis mutant dicarboxylate transport (dct) is one of the classic mutants in the photorespiratory pathway. It requires high CO2 levels for survival. Physiologic and biochemical characterization of dct indicated that dct is deficient in the transport of dicarboxylates across the chloroplast envelope membrane. Hence, re-assimilation of ammonia generated by the photorespiratory cycle is blocked. However, the defective gene in dct has not been identified at the molecular level. Here, we report on the molecular characterization of the defective gene in dct, on the complementation of the mutant phenotype with a wild type cDNA, and on the functional characterization of the gene product, DiT2, in a recombinant reconstituted system. Furthermore, we provide the kinetic constants of recombinant DiT1 and DiT2, and we discuss these data with respect to their functions in ammonia assimilation. Moreover, an analysis of the transcript levels of DiT1 and DiT2 in different C3- and C4-type plant species is presented, and we demonstrate that the substrate specificity of DiT2 from the C4-plant Flaveria bidentis is similar to its counterpart from C3 plants. PMID- 12887584 TI - Transgenic tobacco plants expressing antisense ferredoxin-NADP(H) reductase transcripts display increased susceptibility to photo-oxidative damage. AB - Ferredoxin-NADP(H) reductase (FNR) catalyses the final step of the photosynthetic electron transport in chloroplasts. Using an antisense RNA strategy to reduce expression of this flavoenzyme in transgenic tobacco plants, it has been demonstrated that FNR mediates a rate-limiting step of photosynthesis under both limiting and saturating light conditions. Here, we show that these FNR-deficient plants are abnormally prone to photo-oxidative injury. When grown under autotrophic conditions for 3 weeks, specimens with 20-40% extant reductase undergo leaf bleaching, lipid peroxidation and membrane damage. The magnitude of the effect was proportional to the light intensity and to the extent of FNR depletion, and was accompanied by morphological changes involving accumulation of aberrant plastids with defective thylakoid stacking. Damage was initially confined to chloroplast membranes, whereas Rubisco and other stromal proteins began to decline only after several weeks of autotrophic growth, paralleled by partial recovery of NADPH levels. Exposure of the transgenic plants to moderately high irradiation resulted in rapid loss of photosynthetic capacity and accumulation of singlet oxygen in leaves. The collected results suggest that the extensive photo-oxidative damage sustained by plants impaired in FNR expression was caused by singlet oxygen building up to toxic levels in these tissues, as a direct consequence of the over-reduction of the electron transport chain in FNR deficient chloroplasts. PMID- 12887585 TI - A gene encoding an RNase D exonuclease-like protein is required for post transcriptional silencing in Arabidopsis. AB - Post-transcriptional gene silencing (PTGS) and the closely related phenomenon RNA interference (RNAi) result from the initial endonucleolytic cleavage of target mRNAs, which are then presumed to be completely hydrolyzed by exoribonucleases. To date, no plant genes required for PTGS are known to encode exoribonucleases. The Arabidopsis Werner Syndrome-like exonuclease (WEX) gene encodes an RNase D domain most similar to that in human Werner Syndrome protein (WRN), but lacks the RecQ helicase domain. It is also related to Caenorhabditis elegans mut-7, which is essential for RNAi, PTGS, and transposon activity. We isolated a loss-of function mutant, wex-1, that showed greatly reduced expression of WEX mRNA and early flowering. Although wex-1 did not affect expression of a robust marker for transcriptional gene silencing (TGS), PTGS of a green-fluorescent-protein (GFP) reporter gene was blocked in wex-1 and restored by ectopic expression of WEX, indicating that WEX is required for PTGS but not TGS. Thus, members of the RNase D protein family are required for PTGS in both plants and animals. Interestingly, WEX has been shown to interact with an Arabidopsis RecQ helicase, suggesting that these proteins might comprise a functional equivalent of WRN. PMID- 12887586 TI - Absence of the Lhcb1 and Lhcb2 proteins of the light-harvesting complex of photosystem II - effects on photosynthesis, grana stacking and fitness. AB - We have constructed Arabidopsis thaliana plants that are virtually devoid of the major light-harvesting complex, LHC II. This was accomplished by introducing the Lhcb2.1 coding region in the antisense orientation into the genome by Agrobacterium-mediated transformation. Lhcb1 and Lhcb2 were absent, while Lhcb3, a protein present in LHC II associated with photosystem (PS) II, was retained. Plants had a pale green appearance and showed reduced chlorophyll content and an elevated chlorophyll a/b ratio. The content of PS II reaction centres was unchanged on a leaf area basis, but there was evidence for increases in the relative levels of other light harvesting proteins, notably CP26, associated with PS II, and Lhca4, associated with PS I. Electron microscopy showed the presence of grana. Photosynthetic rates at saturating irradiance were the same in wild type and antisense plants, but there was a 10-15% reduction in quantum yield that reflected the decrease in light absorption by the leaf. The antisense plants were not able to perform state transitions, and their capacity for non-photochemical quenching was reduced. There was no difference in growth between wild-type and antisense plants under controlled climate conditions, but the antisense plants performed worse compared to the wild type in the field, with decreases in seed production of up to 70%. PMID- 12887587 TI - Target genes for OBP3, a Dof transcription factor, include novel basic helix-loop helix domain proteins inducible by salicylic acid. AB - Overexpression of a salicylic-acid (SA)-inducible Arabidopsis DNA binding with one finger (Dof) transcription factor, called OBF-binding protein 3 (OBP3; AtDof3.6), has previously been shown to result in growth defects. In this study, suppressive subtraction hybridization (SSH) was used to isolate genes induced in an OBP3-overexpression line and several putative clones, called OBP3-responsive genes (ORGs), were isolated. The link with the induced expression levels of these genes and OBP3 overexpression was confirmed by analysing additional OBP3 overexpression lines. ORG1 through ORG4 are novel genes, while ORG5 is an extensin gene, AtExt1. While ORG4 has no similarity with other proteins in the database, ORG1 has weak similarity in different regions of the predicted protein with CDC2 and fibrillin. ORG2 and ORG3 share 80% overall identity in their deduced amino acid sequences and contain a basic helix-loop-helix DNA-binding domain, suggesting that ORG2 and ORG3 may be transcription factors. The expression of the ORG1, ORG2 and ORG3 genes was co-regulated under all conditions examined including upregulation by SA and downregulation by jasmonic acid (JA). Fifteen OBP3-silenced lines were generated to further explore the function of OBP3. Although there were no visible phenotypic changes in any of these lines, the expression of ORG1, ORG2 and ORG3 was reduced. Among the ORG genes, ORG1, ORG2 and ORG3 contained the highest number of potential Dof-binding sites in the promoter region, and their expression was significantly increased within 3 h after induction of OBP3 expression using an inducible promoter system, and closely reflected the expression levels of the exogenous OBP3 protein. The results from the gain-of-function and loss-of-function experiments suggest that the ORG1, ORG2 and ORG3 genes are direct target genes of OBP3. PMID- 12887588 TI - The SPA1-like proteins SPA3 and SPA4 repress photomorphogenesis in the light. AB - Suppressor of phyA-105 (SPA1) is a phytochrome A-specific signaling intermediate that acts as a light-dependent repressor of photomorphogenesis in Arabidopsis seedlings. SPA1 is part of a small gene family comprising three genes: SPA1 related 2 (SPA2), SPA1-related 3 (SPA3), and SPA1-related 4 (SPA4). Here, we investigate the functions of SPA3 and SPA4, two very closely related genes coding for proteins with 74% identical amino acids. Seedlings with mutations in SPA3 or SPA4 exhibit enhanced photomorphogenesis in the light, but show no phenotype in darkness. While there are small differences between the effects of spa3 and spa4 mutations, it is apparent that SPA3 and SPA4 function to inhibit light responses in continuous far-red, red, and blue light. Phytochrome A is necessary for all aspects of the spa4 mutant phenotype, suggesting that SPA4, like SPA1, acts specifically in phytochrome A signaling. Enhanced photoresponsiveness of spa3 mutants is also fully dependent on phytochrome A in far-red and blue light, but not in red light. Hence, SPA3 function in red light may be dependent on other phytochromes in addition to phytochrome A. Using yeast two-hybrid and in vitro interaction assays, we further show that SPA3 as well as SPA4 can physically interact with the constitutive repressor of light signaling COP1. Deletion analyses suggest that SPA3 and SPA4, like SPA1, bind to the coiled-coil domain of COP1. Taken together, our results have identified two new loci coding for negative regulators that may be involved in fine tuning of light responses by interacting with COP1. PMID- 12887589 TI - Systemic movement of a tobamovirus requires host cell pectin methylesterase. AB - Systemic movement of plant viruses through the host vasculature, one of the central events of the infection process, is essential for maximal viral accumulation and development of disease symptoms. The host plant proteins involved in this transport, however, remain unknown. Here, we examined whether or not pectin methylesterase (PME), one of the few cellular proteins known to be involved in local, cell-to-cell movement of tobacco mosaic virus (TMV), is also required for the systemic spread of viral infection through the plant vascular system. In a reverse genetics approach, PME levels were reduced in tobacco plants using antisense suppression. The resulting PME antisense plants displayed a significant degree of PME suppression in their vascular tissues but retained the wild-type pattern of phloem loading and unloading of a fluorescent solute. Systemic transport of TMV in these plants, however, was substantially delayed as compared to the wild-type tobacco, suggesting a role for PME in TMV systemic infection. Our analysis of virus distribution in the PME antisense plants suggested that TMV systemic movement may be a polar process in which the virions enter and exit the vascular system by two different mechanisms, and it is the viral exit out of the vascular system that involves PME. PMID- 12887590 TI - Classification and identification of Arabidopsis cell wall mutants using Fourier Transform InfraRed (FT-IR) microspectroscopy. AB - We have developed a novel procedure for the rapid classification and identification of Arabidopsis mutants with altered cell wall architecture based on Fourier-Transform Infrared (FT-IR) microspectroscopy. FT-IR transmission spectra were sampled from native 4-day-old dark-grown hypocotyls of 46 mutants and the wild type treated with various drugs. The Mahalanobis distance between mutants, calculated from the spectral information after compression with the Discriminant Variables Selection procedure, was used for alpha hierarchical cluster analysis. Despite the completely unsupervised nature of the classification procedure, we show that all mutants with cellulose defects appeared in the same cluster. In addition, mutant alleles of similar strength for several unrelated loci were also clustered, which demonstrates the sensitivity of the method to detect a wide array of cell wall defects. Comparing the cellulose deficient cluster with the cluster that contained wild-type controls led to the identification of wave numbers that were diagnostic for altered cellulose content in the context of an intact cell wall. The results show that FT-IR spectra can be used to identify different classes of mutants and to characterize cell wall changes at a microscopic level in unknown mutants. This procedure significantly accelerates the identification and classification of cell wall mutants, which makes cell wall polysaccharides more accessible to functional genomics approaches. PMID- 12887591 TI - A method for profiling classes of plant hormones and their metabolites using liquid chromatography-electrospray ionization tandem mass spectrometry: an analysis of hormone regulation of thermodormancy of lettuce (Lactuca sativa L.) seeds. AB - A highly selective and sensitive method for the simultaneous analysis of several plant hormones and their metabolites is described. The method combines high performance liquid chromatography (HPLC) with positive and negative electrospray ionization-tandem mass spectrometry (ESI-MS/MS) to quantify a broad range of chemically and structurally diverse compounds. The addition of deuterium-labeled analogs for these compounds prior to sample extraction permits accurate quantification by multiple reaction monitoring (MRM). Endogenous levels of abscisic acid (ABA), abscisic acid glucose ester (ABA-GE), 7'-hydroxy-abscisic acid (7'-OH-ABA), phaseic acid (PA), dihydrophaseic acid (DPA), indole-3-acetic acid (IAA), indole-3-aspartate (IAAsp), zeatin (Z), zeatin riboside (ZR), isopentenyladenine (2iP), isopentenyladenosine (IPA), and gibberellins (GA)1, GA3, GA4, and GA7 were determined simultaneously in a single run. Detection limits ranged from 0.682 fmol for Z to 1.53 pmol for ABA. The method was applied to the analysis of plant hormones and hormonal metabolites associated with seed dormancy and germination in lettuce (Lactuca sativa L. cv. Grand Rapids), using extracts from only 50 to 100 mg DW of seed. Thermodormancy was induced by incubating seeds at 33 degrees C instead of 23 degrees C. Germinating seeds transiently accumulated high levels of ABA-GE. In contrast, thermodormant seeds transiently accumulated high levels of DPA after 7 days at 33 degrees C. GA1 and GA3 were detected during germination, and levels of GA1 increased during early post-germinative growth. After several days of incubation, thermodormant seeds exhibited a striking transient accumulation of IAA, which did not occur in seeds germinating at 23 degrees C. We conclude that hormone metabolism in thermodormant seeds is surprisingly active and is significantly different from that of germinating seeds. PMID- 12887592 TI - Development of enhancer trap lines for functional analysis of the rice genome. AB - Enhancer trapping has provided a powerful strategy for identifying novel genes and regulatory elements. In this study, we adopted an enhancer trap system, consisting of the GAL4/VP16-UAS elements with GUS as the reporter, to generate a trapping population of rice. Currently, 31 443 independent transformants were obtained from two cultivars using Agrobacterium-mediated T-DNA insertion. PCR tests and DNA blot hybridization showed that about 94% of the transformants contained T-DNA insertions. The transformants carried, on average, two copies of the T-DNA, and 42% of the transformants had single-copy insertions. Histochemical assays of approximately 1000 T0 plants revealed various patterns of the reporter gene expression, including expression in only one tissue, and simultaneously in two or more tissues. The expression pattern of the reporter gene in T1 families corresponded well with the T0 plants and segregated in a 3 : 1 Mendelian ratio in majority of the T1 families tested. The frequency of reporter gene expression in the enhancer trap lines was much higher than that in gene trap lines reported previously. Analysis of flanking sequences of T-DNA insertion sites from about 200 transformants showed that almost all the sequences had homology with the sequences in the rice genome databases. Morphologically conspicuous mutations were observed in about 7.5% of the 2679 T1 families that were field-tested, and segregation in more than one-third of the families fit the 3 : 1 ratio. It was concluded that GAL4/VP16-UAS elements provided a useful system for enhancer trap in rice. PMID- 12887593 TI - The pathology and pathogenesis of retinal vasculitis. AB - Retinal vasculitis is a rare, but potentially blinding intraocular inflammatory condition with diverse aetiology. Although commonly idiopathic, it has a strong association with systemic inflammatory diseases known to involve other areas of the central nervous system, most notably Behcet's disease, sarcoidosis, systemic lupus erythematosis and multiple sclerosis. This article describes the clinicopathologic features of retinal vasculitis and its visually damaging sequelae, reviewing available human histopathologic studies and work with experimental models to discuss the pathogenesis and immunopathology. Evidence indicates that noninfective retinal vasculitis is an autoimmune condition that may be induced by antecedent infection with microbes cross-reacting with putative autoantigens, influenced by genetic susceptibility of both HLA associations and cytokine polymorphisms. The growing understanding of the cellular mechanisms involved in the effector immune response is already providing a rationale for more specific therapeutic approaches. PMID- 12887594 TI - Reduced rates of axonal and dendritic growth in embryonic hippocampal neurones cultured from a mouse model of Sandhoff disease. AB - Sandhoff disease is a lysosomal storage disease in which ganglioside GM2 accumulates because of a defective beta-subunit of beta-hexosaminidase. This disease is characterized by neurological manifestations, although the pathogenic mechanisms leading from GM2 accumulation to neuropathology are largely unknown. We now examine the viability, development and rates of neurite growth of embryonic hippocampal neurones cultured from a mouse model of Sandhoff disease, the Hexb-/- mouse. GM2 was detected by metabolic labelling at low levels in wild type (Hexb+/+) neurones, and increased by approximately three-fold in Hexb-/- neurones. Hexb-/- hippocampal neurones were as viable as their wild type counterparts and, moreover, their developmental programme was unaltered because the formation of axons and of the minor processes which eventually become dendrites was similar in Hexb-/- and Hexb+/+ neurones. In contrast, once formed, a striking difference in the rate of axonal and minor process growth was observed, with changes becoming apparent after 3 days in culture and highly significant after 5 days in culture. Analysis of various parameters of axonal growth suggested that a key reason for the decreased rate of axonal growth was because of a decrease in the formation of collateral axonal branches, the major mechanism by which hippocampal axons elongate in culture. Thus, although the developmental programme with respect to axon and minor process formation and the viability of hippocampal neurones are unaltered, a significant decrease occurs in the rate of axonal and minor process growth in Hexb-/- neurones. These results appear to be in contrast to dorsal root ganglion neurones cultured from 1-month old Sandhoff mice, in which cell survival is impaired but normal outgrowth of neurones occurs. The possible reasons for these differences are discussed. PMID- 12887595 TI - Apoptotic mitochondrial pathway in neurones and astrocytes after neonatal hypoxia ischaemia in the rat brain. AB - Neuronal apoptosis plays an essential role in early brain development and contributes to secondary neuronal loss after acute ischaemia. Recent studies have provided evidence that caspase-3 is an important downstream event after hypoxia ischaemia in the immature brain, but a minor event in the adult brain. Our investigations have focused on cell populations that expressed apoptotic effectors in the enzymatic death pathway including cytochrome c, caspase-9 and caspase-3. Expression, activation and cellular localization of these proteins were studied using cleavage of fluorogenic substrate and immunohistochemistry in neonatal rat brain after unilateral focal ischaemia. Caspase-3 enzyme activity was elevated in brain homogenate between 6 and 48 h after reperfusion. This activation was preceded by that of caspase-9, between 3 and 24 h. Apoptotic cell death was finally accomplished by poly-ADP-ribose polymerase cleavage, an endogenous caspase-3 substrate. In addition, immunodetection demonstrated that cytochrome c and activated caspase-9 and caspase-3 were expressed not only in the neurones, the primarily affected cells, but also within the astrocytes, which constituted a dense network delineating the infarct. These results suggested that glial injury may promote the formation of cystic lesions such as those observed clinically in the newborn brain. PMID- 12887596 TI - Expression of Tie-2 in human peripheral and autonomic nervous system. AB - Tie-2, a tyrosine kinase receptor, is essential for vascular integrity by regulating cellular adhesion between pericytes and endothelial cells. The aim of this study was to identify sites of expression of Tie-2 other than the vasculature. Tie-2 expression was first detected in human colon by Western blotting and reverse-transcription-polymerase chain reaction (RT-PCR) in tissue extracts. The presence of the Tie-2 mRNA and protein was detected by immunohistochemistry and in situ hybridization in cells of the colon myenteric and submucosal plexus, in both neuronal and Schwann cells. Tie-2 protein was also found in the nervous system of the female urogenital tract. In the human sciatic nerve and schwannoma, RT-PCR, Western blotting and immunohistochemistry analysis further confirmed the presence of Tie-2 mRNA and protein in non-autonomic peripheral nervous tissue. In conclusion, using several approaches and tissues we have demonstrated the presence of Tie-2 in human peripheral and autonomic nervous tissue, suggesting a role for Tie-2 in neural tissue. Thus, attempts to disrupt the tumour vessels by manipulation of the Tie-2 system in tumours may result in side-effects in peripheral nerves. PMID- 12887597 TI - AMOG/beta2 and glioma invasion: does loss of AMOG make tumour cells run amok? AB - The beta2 subunit of Na,K-ATPase, initially described as adhesion molecule on glia (AMOG), has been shown to mediate neurone-astrocyte adhesion as well as neural cell migration in vitro. We have investigated the expression of AMOG/beta2 in human gliomas and its effect on glioma cell adhesion and migration. Compared to normal astrocytes of human brain, AMOG/beta2 expression levels of neoplastic astrocytes were down-regulated in biopsy specimens and inversely related to the grade of malignancy. One rat and four human glioma cell lines showed complete loss of AMOG. To investigate the function of AMOG/beta2, its expression was re established by transfecting an expression plasmid into AMOG/beta2-negative C6 rat glioma cells. In vitro assays revealed increased adhesion and decreased migration on matrigel of AMOG/beta2-positive cells as compared to their AMOG/beta2-negative counterparts. We conclude that increasing loss of AMOG/beta2 during malignant progression parallels and may underlie the extensive invasion pattern of malignant gliomas. PMID- 12887598 TI - Relationship of Nef-positive and GFAP-reactive astrocytes to drug use in early and late HIV infection. AB - Reactive astrocytosis is a well-documented feature of HIV encephalitis (HIVE), but it is unclear whether restricted infection of astrocytes contributes to this phenomenon. In addition, the part played by reactive and/or infected astrocytes in AIDS-related dementia is not fully understood. In this study of patients at different stages of the human immunodeficiency virus (HIV) infection, who had been treated at most with one antiretroviral drug, reactive astrocytes were identified by immunopositivity for glial fibrillary acidic protein (GFAP) and infected astrocytes by positivity for HIV Nef protein. Results were compared for drug-using AIDS patients with (n=9) and without (n=7) HIVE, for presymptomatic HIV-positive drug users (n=12) and for control HIV-negative subjects (n=20), including a group who used drugs (n=10). GFAP-reactive astrocytes in both grey and white matter were significantly more numerous in HIVE subjects than in each of the other groups but did not correlate with viral load. Nef-positive astrocytes were confined to HIVE cases and to white matter, but were numerous in only one subject who was treatment-naive. Nef-positive microglia were identified in all HIVE cases and in occasional AIDS and presymptomatic subjects who did not have HIVE. The results suggest that astrocytes may form an additional viral reservoir in late HIV infection and may contribute to HIVE. However, the number of GFAP-positive astrocytes was neither increased in pre-AIDS nor in drug abuse, in contrast with microglia which we have shown previously to be up-regulated in both states. PMID- 12887599 TI - The allograft inflammatory factor-1 in Creutzfeldt-Jakob disease brains. AB - The allograft inflammatory factor-1 (AIF-1) is a 17-kDa IFN-gamma inducible Ca(2+)-binding EF-hand protein that is encoded within the HLA class III genomic region and is involved in immune dysfunction and smooth muscle cell activation. We used immunohistochemistry double labelling experiments to analyse the spatial distribution and cell-type-specific localization of AIF-1 in the brains of patients who died as a result of sporadic Creutzfeldt-Jakob disease (CJD) and neuropathologically unaltered controls. Significantly more AIF-1 immunoreactive macrophages/microglial cells and, interestingly, neurones were observed in CJD patients compared to controls. Western blotting confirmed more prominent AIF-1 immunoreactive bands of approximately 50 kDa in four CJD patients compared to three controls. Chaotropic SDS-PAGE of the recombinant AIF-1 resulted in almost complete reduction of the 50 kDa band and mass spectrometry revealed only AIF-1 specific tryptic protein fragments suggesting that trimerized AIF-1 is the predominant form in vivo. Finally, we analysed mechanisms of neuronal AIF-1 induction. Following H2O2 challenge, a model of general cell stress, we observed the gradual induction of AIF-1 and, more interestingly, release to the supernatant of SKNSH neurones. Parallel reverse transcriptase polymerase chain reaction and sequencing was used to confirm AIF-1 mRNA expression. PMID- 12887600 TI - Glycohistochemical characterization of vascular muscle cell destruction in CADASIL subjects by lectins, neoglycoconjugates and galectin-specific antibodies. AB - CADASIL (Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy) is a type of small-artery stroke and vascular dementia inducing pathology of the brain. In order to explain the molecular mechanisms behind the alterations to the blood vessels in CADASIL subjects, we scrutinized the expression of glycan and glycan-binding sites in the wall of vessels taken from five such subjects (vs. five control subjects matched for age and sex). Specimens were taken from the brain, heart, kidney, liver and lung. Although the main vessel lesions were observed in the tissues depending on the blood-brain barrier, alterations to systemic vessels were also observed despite the absence of any symptoms. The histochemical expression of a panel of 10 biotinylated neoglycoconjugates [Gal-beta(1-4)-D-Glc, Galbeta(1-3)GalNAc, alpha-D-GalNAc, beta D-GalNAc, GalNAcalpha(1-3)-D-GalNAcalpha, GalNAcalpha(1-3)-D-GalNAcbeta, beta-D Glc, alpha-D-Man, l-Fucose and D-Glcalpha(1-4)-D-Glc], eight plant lectins (PNA, MAA, SNA, DBA, WGA, ConA, GNA and UEA-1) and two antigalectin antibodies was monitored by means of semiquantitative and quantitative computer-assisted microscopy. The data show the altered histochemical binding of plant lectins, such as UEA-1 and ConA, in the vessel walls of CADASIL subjects. The present work, based upon staining by a panel of neoglycoconjugates, provides a biochemical characterization of the alteration of vessel walls in the brain compared to other organs including the heart, kidney, lung and liver in CADASIL as opposed to control subjects. These glycohistochemical results suggest a functional relevance of protein-carbohydrate interactions in this disease. PMID- 12887601 TI - Dural haemorrhage in nontraumatic infant deaths: does it explain the bleeding in 'shaken baby syndrome'? Geddes JE et al. A response. PMID- 12887604 TI - In vivo release of oxytetracycline from a biodegradable controlled-release gel injected subcutaneously in Japanese quail (Coturnix coturnix japonica). AB - A long-acting, biodegradable, controlled-release formulation of oxytetracycline (CR-OTC) was evaluated in 18 adult Japanese quail (Coturnix coturnix japonica) following a single subcutaneous (s.c.) injection. Prior to characterizing the release of oxytetracycline (OTC) from the CR-OTC, the pharmacokinetic parameters of intravenously (i.v.) administered OTC were determined. Concentrations of free OTC were measured using a bioassay. The plasma concentration-time profile of OTC after a single i.v. injection at 20 mg/kg was best fit to an open two compartmental model, with the following pharmacokinetic parameters: area under the curve (AUC) = 36.72 mg. h/L, terminal elimination half-life = 2.34 h, clearance (Cl) = 0.545 L/kg/h. Plasma [OTC] was >1.0 micro g/mL for at least 4 h following i.v. injection. The CR-OTC gel was well tolerated at a dosage of 1500 mg/kg s.c. Plasma [OTC] rose to >1.0 micro g/mL within 24 h; it remained >1.0 micro g/mL for at least 10 days in all birds sampled at that time point (n = 9) and for at least 18 days in two of nine birds. Using a deconvolution technique, it was determined that approximately 54.8% of the administered OTC was released from the CR-OTC over the 45-day observation period. This long-acting, biodegradable controlled-release OTC formulation may have potential for the treatment of chlamydophila infections and other OTC-sensitive bacteria in Japanese quail, however further studies are necessary to determine its safety and clinical application. PMID- 12887605 TI - Clinical efficacy of trimethoprim/sulfadiazine and procaine penicillin G in a Streptococcus equi subsp. zooepidemicus infection model in ponies. AB - Tissue chambers, implanted subcutaneously on both sides of the neck in eight ponies, were inoculated with Streptococcus equi subsp. zooepidemicus in order to compare the clinical efficacy of trimethoprim/sulfadiazine (TMP/SDZ) and penicillin G treatment in a purulent infection. The TMP/SDZ treatment consisted of one intravenous (i.v.) injection of 5 mg/kg TMP and 25 mg/kg SDZ and the same dose of TMP/SDZ per os (p.o.), both given 20 h after inoculation. The oral dose was then repeated every 12 h for 21 days. The penicillin treatment consisted of one i.v. injection of 20 000 IU/kg sodium penicillin G and intramuscular (i.m.) injection of 20 000 IU/kg procaine penicillin G, both given 20 h after infection. The i.m. dose was then repeated every 24 h for 21 days. Eight ponies, each with two tissue chambers, were used in a cross over design; in the first experiment the left tissue chamber (TC) was infected and in the second experiment the right. TMP/SDZ treatment resulted in a limited reduction of viable bacteria in the TC but did not eliminate the infection, resulting in abscessation in 10-42 days in all eight ponies. However, penicillin treatment eliminated the streptococci in seven of eight ponies, and only one pony suffered abscessation on day 10. This constitutes a significantly better efficacy of the penicillin treatment in this model. The most probable cause of the failure of TMP/SDZ to eliminate the streptococci is inhibition of the action of TMP/SDZ in the purulent TCF. Therefore, TMP/SDZ should not be used to treat purulent infections in secluded sites in horses. PMID- 12887606 TI - Comparative effects of fluoroquinolones on subsets of T lymphocytes in normothermic and hyperthermic mice. AB - The subsets of T lymphocytes in thymus, spleen and mesenteric lymph nodes were investigated in normothermic and hyperthermic mice treated with fluoroquinolones administered orally six times at 24 h intervals at doses of 15 or 75 mg/kg (flumequine, norfloxacin and ciprofloxacin) and 5 or 25 mg/kg (enrofloxacin). It has been found that fluoroquinolones can modulate CD3+, CD4+ and CD8+ marker expression on thymocytes, splenocytes and lymphocytes of mesenteric lymph nodes. Flumequine (15 mg/kg) decreased the percentage of immature CD4+CD8+ thymic cells and increased the percentage of mature CD4+ and CD8+. When the dose of flumequine was increased to 75 mg/kg a reduction in the maturation of thymocytes was observed. Administration of flumequine, norfloxacin and ciprofloxacin, irrespective of doses applied, increased the percentages of CD3+ splenocytes of CD4+ spleen cells. Exposure to enrofloxacin decreased the percentage of T helper inducer cells. Flumequine and ciprofloxacin augmented the percentage of CD3+ mesenteric lymph node cells and increased the percentage of CD8+ cells. In contrast, norfloxacin and enrofloxacin decreased the percentage of CD3+ mesenteric lymph node cells and the percentage of CD4+ cells. Lipopolysaccharide (LPS) from E. coli (25 micro g/mouse) increased the percentage of single-positive CD4+ thymocytes, but did not affect the percentage of CD3+, CD4+ and CD8+ splenocytes and mesenteric lymph node cells. Flumequine and ciprofloxacin administered to mice pior to LPS potentiated its stimulant effect on the maturation of thymic cells ( increased percentage of mature CD4+ and CD8+ thymocytes). Pre-treatment with norfloxacin or enrofloxacin either reduced or did not modify the stimulant effect of LPS on maturation of thymic cells. Flumequine, norfloxacin, enrofloxacin and ciprofloxacin administered prior to LPS decreased the percentage of CD8+ splenocytes and increased the percentage of CD4+ spleen cells. Norfloxacin and ciprofloxacin at a dose of 75 mg/kg reduced the percentage of CD8+ mesenteric lymph node cells in hyperthermic mice. Pretreatment with norfloxacin at a dose of 15 mg/kg augmented the percentage of mesenteric lymph node cells. It was concluded that the modulating effects of fluoroquinolones depends on the chemial structure of drugs, dose administered as well as immunologic status. PMID- 12887607 TI - Single dose pharmacokinetics of piroxicam in cats. AB - Piroxicam (PIRO) is a nonsteroidal anti-inflammatory drug (NSAID) recognized for its value as a chemopreventative and anti-tumor agent. Eight cats were included in this study. PIRO was administered in a single oral (p.o.) and intravenous (i.v.) dose of 0.3 mg/kg. The study was designed as a randomized complete crossover with a 2-week washout period. Serial blood samples were collected after each dose and plasma was analyzed for PIRO. Pharmacokinetic parameters of PIRO were determined using noncompartmental analysis. PIRO is well absorbed in the cat with a median bioavailability (F) of 80% (range 64-124%). The median i.v. t1/2 was 12 h (range 8.6-14 h). The median Cmax was 519 ng/mL with a corresponding Tmax of 3 h. PIRO appears to be rapidly absorbed following p.o. administration in cats with a higher Cmax and AUC than in dogs. PMID- 12887608 TI - Effect of alkalinization on the local analgesic efficacy of ketamine in the abaxial sesamoid nerve block in horses. AB - The objective of this study was to determine the effects of the alkalinization on the local analgesic efficacy of 1% ketamine in the abaxial sesamoid nerve block in horses. Thirty-six mature healthy horses were randomly assigned to four groups for the following treatments; an abaxial sesamoid block with 5 mL of saline solution (control saline); an abaxial sesamoid block with 5 mL of a solution containing 1% ketamine (KETs 1%); an abaxial sesamoid block with 5 mL of a solution containing saline solution and 0.5 mEq of sodium bicarbonate (control bicarbonate); and an abaxial sesamoid block with 5 mL of a solution containing 1% ketamine and 0.5 mEq of sodium bicarbonate (KETb 1%). All blocks were performed in one randomly selected front leg. To determine analgesia, hoof withdrawal from thermal stimulus from radiant heat lamp was assessed. Before each block, the hoof withdrawal reflex latency (HWRL) (time between lamp illumination and withdrawal of the hoof) was determined; after the block, local analgesic effects were determined using the heat lamp at 2 and 5 min after the injection and then every 5 min for 1 h. In KETs 1% group, there were significant increases in HWRL between basal values and values from 2 to 10 min after an abaxial sesamoid block. In KETb 1% group, significant increases in HWRL was collected between the basal value and values from 2 to 25 min following an abaxial sesamoid block. In KETs 1% group, of the nine horses, four had an abaxial sesamoid block that was unsuccessful. However, in KETb 1% group, only one of the nine horses had an abaxial sesamoid nerve block that was unsuccessful. The alkalinization of a 1% ketamine solution produced a more consistent and persistent local analgesia in horses when compared with 1% ketamine solution alone. PMID- 12887609 TI - Pharmacokinetics and sedative effects of intramuscular medetomidine in domestic sheep. AB - Intramuscular (i.m.) administration of medetomidine (MED) may avoid the severe pressor effects caused by peripheral actions of MED associated with intravenous (i.v.) dosing. The purpose of this study was to determine the pharmacokinetics, the time course of sedation and occurence of hypoxaemia after i.m. administration of MED in domestic sheep. The MED was injected i.m. at a dose of 30 micro g/kg in nine domestic sheep. Blood was sampled at 0, 5, 10, 20, 30, 40, 60, 120, 180, 240, 360 and 600 min after MED. Sedation was assessed and arterial blood samples were taken before and 35 min after MED application. Mean (SD) pharmacokinetic parameters of i.m. MED were: absorption half-life: 13.2 (7.5) min; terminal half life: 32.7 (14.9) min; time to peak concentration: 29.2 (8.9) min; peak concentration: 4.98 (1.89) ng/mL; volume of distribution: 3.9 (2.4) l/kg; total body clearance: 81.0 (21.5) mL/(min kg). Peak sedation occurred between 30 and 40 min after injection of MED. The degree of sedation correlated with individual plasma concentrations (rS: 0.926). One animal became hypoxaemic (PaO2 = 54.1 mmHg). PMID- 12887610 TI - Differential effects of phosphodiesterase inhibitors on platelet activating factor (PAF)- and adenosine diphosphate (ADP)-induced equine platelet aggregation. AB - Compounds that activate adenylate cyclase, increasing intracellular cyclic adenosine monophosphate (cAMP), inhibit equine platelet aggregation. Cyclic AMP is broken down by phosphodiesterase (PDE) and, in the present study, the effects of theophylline, a nonselective PDE inhibitor, and selective inhibitors of PDE isoenzymes PDE3, PDE4 and PDE5, on equine platelet aggregation in response to platelet activating factor (PAF) and adenosine diphosphate (ADP) have been examined. Theophylline and the PDE3 inhibitors, trequinsin and quazinone, inhibited both PAF and ADP-induced aggregation in a concentration dependent manner. The inhibition of PAF-induced aggregation was, however, significantly greater than that of the response to ADP. The inhibitory effects of theophylline and the PDE3 inhibitors on ADP- but not PAF-, induced aggregation were reversed by addition of the calcium ionophore, A23187. Rolipram and zaprinast, inhibitors of PDE4 and PDE5, respectively, had no effect on either PAF- or ADP-induced aggregation. These results demonstrate that inhibition of aggregation caused by PAF or ADP can be achieved by selective inhibition of PDE3 but suggest that there may be agonist-specific differences in the intracellular signalling pathways that regulate equine platelet aggregation. PMID- 12887611 TI - Pharmacologic identification of putative D1 dopamine receptors in feline kidneys. AB - The presence of dopamine (DA) receptors in feline kidneys is a matter of contention. Radioligand binding and Western blotting studies were employed to determine whether DA receptors are present in feline kidneys. The pharmacologic profile of the selective D1-receptor antagonist [3H]-SCH 23390 was studied in renal cortical membrane preparations from cats by conducting saturation binding isotherm and competitive binding experiments. [3H]-SCH 23390 bound to feline renal cortical membranes in a manner consistent with labeling of a D1-like receptor. The binding profile revealed a single site D1-like or D1 receptor in the feline renal cortex with a Kd = 7.8 +/- 1.0 nmol/L and Bmax = 76.5 +/- 19.5 fmol/mg. Competitive binding studies for [3H]-SCH 23390 against unlabeled agonists yielded the following Ki values and rank order of competition: SKF38393 (Ki = 0.47 +/- 0.26 micro m) > fenoldopam (Ki = 3.12 +/- 1.1 micro m) > DA (Ki = 933.1 +/- 1.6 micro m). Competitive binding studies for [3H]-SCH-23390 against unlabeled antagonists yielded the following rank order of competition: SCH 23390 (Ki = 1.97 +/- 0.81 micro m) > spiperone (Ki = 3.79 +/- 0.79) > metoclopramide (Ki = 4.26 +/- 2.4 micro m). Western blot analysis with anti-DA D1 receptor antibodies detected a single band with Mr of 74 kDa corresponding to a D1 DA receptor. These results suggest that a putative D1-like or D1 receptor exists in feline kidneys different from those previously identified in rat, dog or human kidneys. PMID- 12887612 TI - The effect of food or water deprivation on paracetamol pharmacokinetics in calves. AB - This study investigated the effect of food or water deprivation on the pharmacokinetics of paracetamol in 30 Holstein-Friesian preruminant calves (10 controls, 10 food withheld and 10 water-deprived) aged 24-25 days. Control calves were given paracetamol at 24-25 days and again at 28-29 days of age. In the food withheld and water-deprived calves paracetamol studies were performed before and after 4 days of food or water deprivation. In the control group there were no significant differences in pharmacokinetic parameters for paracetamol in 24-25 and 28-29-day-old calves. Witholding food for 4 days was associated with an increase in the mean residence time (MRT) of paracetamol (P < 0.01). When food was withheld total body clearance (ClB) of paracetamol was significantly decreased (P < 0.05). The volume of distribution (Vss) was not significantly altered. Similarly, water deprivation was associated with a significant increase in MRT and significant decrease in ClB of paracetamol (P < 0.01). The Vss was not significantly altered. Food or water deprivation also influenced the formation of major metabolites (glucuronide and sulphate) of paracetamol. It is concluded that food or water deprivation may impair the elimination drugs that undergo metabolism by UDP-glucuronyltransferase and sulphotransferase in cattle. PMID- 12887613 TI - Stereospecific biotransformation of albendazole in mouflon and rat-isolated hepatocytes. AB - The anthelmintic albendazole (ABZ) undergoes a two-step oxidation resulting first in the formation of chiral albendazole sulfoxide (ABZSO) followed by its transformation to albendazole sulfone (ABZSO2) in many farm and laboratory animal species. Although cloven-hoofed game are also treated with ABZ, limited information concerning ABZ biotransformation in these species is available. The present study focused on in vitro ABZ sulfoxidation in hepatocytes from wild sheep-mouflon (Ovis musimon) and comparison of ABZ sulfoxidation in mouflon and rat (Rattus norvergicus) hepatocytes. ABZ was used as a substrate for primary cultures of mouflon and rat hepatocytes. Time-dependent stereospecific consumption of ABZSO and ABZSO2 formation has been investigated. The metabolites were determined by high-performance liquid chromatography with both achiral and chiral stationary phases. Although total-ABZSO formation did not significantly differ between mouflon and rat, after separation of the (+)-ABZSO and (-)-ABZSO enantiomers a significant difference between species was found. The enantiomeric ratio of (+)/(-)-ABZSO in mouflon hepatocytes was 2.8-3.8, while rat hepatocytes biotransformed ABZ to almost racemic ABZSO, with an enantiomeric ratio of 1.0 1.1. The ratio were similar for two concentrations of substrate used and stable over several time intervals. The formation of ABZSO2 was more extensive in rat (approximately five times) than in mouflon hepatocytes. PMID- 12887614 TI - Pharmacokinetics of enrofloxacin and its active metabolite ciprofloxacin in cows following single dose intravenous administration. PMID- 12887615 TI - Pharmacokinetics in plasma of ivermectin after its oral (solution and tablets) administration to sheep. PMID- 12887616 TI - The effect of solute lipophilicity on penetration through feline skin. PMID- 12887618 TI - Concept and realization of a multimedia program for veterinary physiology. AB - A multimedia program, a non-linear system which supplements the traditional, linear way of learning (e.g. with a text book), was developed. Non-linear systems allow the direct use of information in a free sequence. A problem of these systems is the danger that the reader becomes lost in detail. Therefore, a program was constructed which tries to combine the properties of linear and non linear teaching systems. Student groups were continuously involved during all stages of the project. The program is organized in several levels. These are: (1) choice of chapters; (2) table of contents of one chapter; (3) summary; (4) text and (5) pictures, animations, simulations and video sequences. The program can be used in different ways. In the linear mode, the reader selects via action words different levels of the program. The user can read either only on the level of the summaries or on detailed information (text, pictures, simulations and videos). In the non-linear mode the user jumps via menu items or buttons to selected pictures, simulations or a searched term. In this way, more than 3500 screen pages are integrated into a multimedia program for veterinary physiology. The program is written in TOOLBOOK, and can be used on a personal computer with Windows 95 or higher. PMID- 12887619 TI - Analysis of ELA-DQB exon 2 polymorphism in Argentine Creole horses by PCR-RFLP and PCR-SSCP. AB - The second exon of equine leucocyte antigen (ELA)-DQB genes was amplified from genomic DNA of 32 Argentine Creole horses by PCR. Amplified DNA was analysed by PCR-restriction fragment length polymorphism (RFLP) and PCR-single-strand conformation polymorphism (SSCP). The PCR-RFLP analysis revealed two HaeIII patterns, four RsaI patterns, five MspI patterns and two HinfI patterns. EcoRI showed no variation in the analysed sample. Additional patterns that did not account for known exon 2 DNA sequences were observed, suggesting the existence of novel ELA-DQB alleles. PCR-SSCP analysis exhibited seven different band patterns, and the number of bands per animal ranged from four to nine. Both methods indicated that at least two DQB genes are present. The presence of more than two alleles in each animal showed that the primers employed in this work are not specific for a unique DQB locus. The improvement of this PCR-RFLP method should provide a simple and rapid technique for an accurate definition of ELA-DQB typing in horses. PMID- 12887620 TI - Fibrosarcomas at presumed sites of injection in dogs: characteristics and comparison with non-vaccination site fibrosarcomas and feline post-vaccinal fibrosarcomas. AB - Fifteen fibrosarcomas, surgically excised from presumed sites of injection in dogs, and 10 canine fibrosarcomas excised from sites not used for injection were histologically and immunohistochemically compared with 20 feline post-vaccinal fibrosarcomas. Canine fibrosarcomas from presumed injection sites were of grade I (3), of grade II (4) and grade III (8). Two fibrosarcomas from non-injection sites were of grade I, four of grade II and four of grade III. Feline samples were classified as grade I (2), grade II (4) and grade III (14). All fibrosarcomas from presumed injection sites of both species showed lymphocytic inflammatory infiltration located at the tumour periphery, while two canine fibrosarcomas from non-injection sites showed perivascular inflammatory infiltration within the neoplasm. All samples were immunohistochemically examined for vimentin, smooth muscle actin, muscle specific actin and desmin expression. All tumours were positive for vimentin. Ten canine fibrosarcomas from presumed injection sites and all feline samples contained cells consistent with a myofibroblastic immunophenotype. Aluminium deposits were detected in eight canine fibrosarcomas from presumed injection sites and 11 feline post-vaccinal fibrosarcomas by the aurintricarboxylic acid method. The present study identifies distinct similarities between canine fibrosarcomas from presumed injection sites and feline post-vaccinal fibrosarcomas, suggesting the possibility of the development of post-injection sarcomas not only in cats, but also in dogs. PMID- 12887621 TI - Degenerative myelopathy associated with cobalamin deficiency in a cat. AB - A severe myelopathy was observed in a 9-year-old neutered male cat with a clinical history of chronic pancreatitis associated with deficiency of serum cobalamin and folates concentrations, and progressive spinal ataxia. The spinal cord lesions mainly involved the dorsal columns of the caudal cervical and cervico-thoracic segments, and were characterized by diffuse vacuolated myelin sheaths and axonal degeneration, marked gliosis, fibrosis and presence of gitter cells. The pancreas showed severe atrophy of the exocrine tissue, periductular fibrosis and infiltration of inflammatory cells, consistent with chronic interstitial pancreatitis. This condition can be accountable for cobalamin deficiency, as the pancreas is the only source of intrinsic factor in cats. The spinal cord lesions in the cat of this report resembled the subacute combined degeneration of the spinal cord described in human beings with cobalamin deficiency and hence a similar pathogenetic mechanism is hypothesized. PMID- 12887622 TI - Plasma profiles of ivermectin in horses following oral or intramuscular administration. AB - A study was undertaken in order to evaluate and compare ivermectin's (IVM) plasma disposition kinetic parameters after oral or intramuscular (IM) administration in horses. Ten clinically healthy adult horses, weighing 380-496 kg body weight (BW), were allocated to two experimental groups of five horses. Group I, was treated with an oral paste formulation of IVM at the manufacturer's recommended dose of 0.2 mg/kg BW. Group II, was treated IM with an injectable 1% formulation of IVM at a dose of 0.2 mg/kg BW. Blood samples were collected by jugular puncture at different times between 0.5 h and 75 days post-treatment. After plasma extraction and derivatization, samples were analysed by high-performance liquid chromatography with fluorescence detection. A computerized kinetic analysis was performed, and data were compared using the Wilcoxon signed rank test. The parent molecule was detected in plasma between 30 min and either 20 (oral) or 40 (IM) days post-treatment. Significant differences were found for the time corresponding to peak plasma concentrations (tmax) and for absorption half life. Peak plasma concentrations (Cmax) of 51.3 +/- 16.1 ng/ml (mean +/- SD) were obtained after oral administration and of 31.4 +/- 6.0 ng/ml for the IM route. The values for area under concentration-time curve were 137.1 +/- 35.9 ng day/ml for the group treated orally, and 303.2 +/- 4.3 ng day/ml for the IM treated group. The mean plasma residence times were 4.2 +/- 0.4 and 8.9 +/- 0.7 days for oral and IM-treated groups, respectively. The results of this study show that the route of administration considerably affects the disposition of IVM. A significant difference in bioavailabilty and half-life of elimination of IVM was observed after IM administration compared with oral administration. A close relationship between pharmacokinetic profiles and the clinical efficacy of IVM was established. PMID- 12887623 TI - Blood type A and B frequencies in Turkish Van and Angora cats in Turkey. AB - Blood typing of domestic cats has been performed in domestic and purebred cats in various parts of the world and is important in clinical practice in order to prevent neonatal isoerythrolysis (NI) and acute haemolytic transfusion reactions. Prevalence of blood types vary greatly between breeds of cats. Turkish Van and Angora cats are different breeds that originated in geographically distinct regions of Turkey. The present survey determined the frequency of blood types in these Turkish pedigreed cats in Turkey. Ethylenediaminetetraacetic-acid anti coagulated blood of a total of 113 Turkish Van and Angora cats were examined for blood typing using a slide and tube agglutination assay. Of the 85 Van cats surveyed, 40% had type A, and 60% had type B blood. Of the 28 Turkish Angora cats, 53.6% had type A, and 46.4% had type B blood. No type AB cats were found between both breeds. There was no significant association between blood types and gender of both Angora and Van cats or eye colours of Van cats (P > 0.05). Although these are limited surveys, the overall prevalence of type B cats in these two breeds was very high compared with the results of previous studies worldwide. It appears likely that blood type incompatibilities responsible for feline NI and transfusion reactions are occurring in these breeds. The risk of transfusion incompatibility in Turkish Angora and Van cats was 46.4 and 60%, respectively. It is therefore strongly recommended to breeders and clinicians that blood typing be performed prior to breeding and transfusing cats. PMID- 12887624 TI - Effects of hyperosmolar ionic and low osmolar non-ionic contrast media on coagulation times and some blood parameters in dogs: an in vivo study. AB - The purpose of this study is to evaluate the effects of hyperosmolar ionic contrast media (CM) (diatrizoate) and low osmolar non-ionic CM (iohexol and ioxilan) on coagulation time and some blood parameters in dogs in vivo. The animals were divided into three groups in equal numbers. The dogs in groups I, II and III received diatrizoate, iohexol and ioxilan at the dose of 700 mgI/kg intravenously (IV) as a bolus, respectively. Administration of contrast media and blood samples were collected from vena cephalica antebrachii prior to CM administration and thereafter at 3, 15, 30, 60, 90 and 180 min and 24 h to measure the coagulation factors [activated partial thromboplastin time (APTT), prothorombin time (PT), fibrinogen and fibrinogen degradation products] and some other blood parameters [red blood cells, platelet, white blood cells, haematocrit (Ht) and haemoglobin (Hb)]. While a statistically significant decrease was observed on APTT at 15 min in group III, no significant differences were found in groups I and II. All the groups had insignificant alterations for PT, fibrinogen and fibrinogen degradation product, following CM administration. Significant decreases were observed for platelet at 3 min in all groups. This decrease was also significant at 15- and 30- min intervals in group I. There were significant decreases for erythrocytes, Ht and Hb measurements within 30 min, and no significant alterations were observed for leucocytes within 60 min in all groups compared with baseline values. No differences were observed with regard to coagulation times and some blood parameters as far as long-lasting and major effects of each CM are concerned. PMID- 12887625 TI - Response of dogs to short-term infusions of carbohydrate- or lipid-based parenteral nutrition. AB - Parenteral nutrition (PN) is used to support intensive care patients. The risk for adverse metabolic effects depends on the composition of infused solutions and the duration of application. The present study in dogs compares metabolic and endocrine effects of two infusion solutions, with either triglycerides or glucose being the major energy sources, administered in a comparatively short infusion period (10 h/day). PN was administered for 9 days to two groups of five adult dogs to meet energy maintenance requirements. In group PN-LIP 61% of the total energy was derived from lipids and 22% from carbohydrates, compared with 21 and 62% in group PN-GLUC. Among routine haematology and clinical chemistry the plasma levels of glucose, triglycerides, insulin, insulin-like growth factor-I (IGF-I), glucagon, 3,5,3'-triiodothyronine and thyroxin were measured in non-infused dogs and at 2, 4, 6, and 8 h after the start of infusion at days 2 and 8 of the study. Infusions protocols did not cause gross metabolic aberrations. During the actual infusions glucose, triglyceride and insulin concentrations were elevated, each depending on the infusion solution. Concentrations of IGF-I, glucagon, 3,5,3' triiodothyronine, thyroxin and cortisol did not change significantly. In conclusion short infusion periods of 10 h per day were tolerated by healthy dogs without adverse signs, which could improve practicability of PN also in clinical cases. PMID- 12887626 TI - Comparison of the techniques of evaluation of urine dilution/concentration in the dog. AB - The objective of this study was to evaluate the quality of the measurement of dog urine dilution/concentration by comparing osmolality with three methods of specific gravity (USG) measurement, i.e. weighing, refractometry and test strips. In unselected urine samples from 182 dogs there was a better agreement between osmolality and USG determination by refractometry (r = 0.92) than by weighing (r = 0.82) or by test strips (r = 0.27). There was an almost linear relationship between osmolality and USG: osmolality (mOsm/kg) = 36646(34318/38974) x (USGref - 1) + 25(-39/88); calculated osmolality differed from measured osmolality by more than 500 mOsm/kg in only 8 of 181 samples. There was a good agreement between USG determination by weighing and refractometry: USGref = 1.000(0.905/1.095) x USGweighing - 0.0004(-0.0019/0.0027), with a moderate bias. Only 12% of the differences between the two methods exceeded 0.010. Test strip assessment of USG was unreliable because of systematic underestimation and should not be used for dog urine. Refractometry is the best technique for routine evaluation of urine concentration/dilution when osmometry is not available. PMID- 12887627 TI - Effects of inter- and intra-observer variability on echocardiographic measurements in awake cats. AB - The objective of this study was to determine intra- and inter-observer variability of echocardiographic measurements in awake cats. Four observers with different levels of experience in echocardiography performed 96 echocardiographic examinations in four cats on four different days over a 3-week period. The examinations were randomized and blinded. The maximum within-day and between-day CV values were 17.4 and 18.5% for inter-ventricular septal thickness in diastole, 18.7 and 22.6% for left ventricular free-wall thickness in diastole, 9.8 and 14.9% for left ventricular end-diastolic diameter, 20.8 and 15.2% for left ventricular end-systolic diameter, and 21.2 and 18.4% for left ventricular shortening fraction. The maximum within-day CV values were most often associated with the least competent observer (i.e. the graduate student) and, the minimum CV values with the most competent observer (i.e. the associate professor in cardiology). A significant interaction between cat and observer was also evidenced. Thus, the most competent observer could not be replaced by any of the other observers. PMID- 12887628 TI - Mental health promotion and suicide prevention in Iraq after regime change. PMID- 12887629 TI - Consumer participation in mental health services: looking from a consumer perspective. AB - Widespread changes to the structure and delivery of mental health services have effected considerable change in the role of the service user or consumer. The view of consumers of mental health services as passive recipients of care and treatment is gradually undergoing a significant shift, in light of an increasing expectation that consumers be provided with opportunities to become actively involved in all aspects of their care. Consumer participation is now broadly reflected in government policy; however, to date there has been little exploration of the extent to which the policy is being realized in practice. To provide a greater understanding of these experiences and opinions, in-depth interviews were conducted with consumers of mental health services (n = 15). The interview transcripts were analysed through the identification and explication of major themes. The findings reinforce the need to view consumers as heterogeneous and respond to individual needs and interests regarding consumer participation. Despite variations in experience there is a clear need to develop mechanisms to support consumer involvement and to influence the attitudes of health professions to become more valuing of a consumer perspective. Nurses are in an ideal position to lead this process. PMID- 12887630 TI - Travelling through the system: the lived experience of people with borderline personality disorder in contact with psychiatric services. AB - This study adopted a lived experience approach utilizing a broad research question to ask 'How do people with borderline personality disorder (BPD) experience their contact with mental health services?'. Unstructured interviews were conducted with seven participants from one mental health trust. A grounded theory methodology was adopted to develop a theory of how people with BPD conceptualized their contact with psychiatric services and what strategies they adopted to negotiate their way through them. The study found that people with BPD valued their contact with psychiatric services despite negative experiences and encountering negative staff attitudes. It also found that relationships with others were vital in containing their distressing emotions despite their difficulty in trusting others. Overcoming these paradoxes was achieved by consistent long-term involvement with experienced staff, containing relationships, encouraging the participants to contribute to their care through the structure of the care programme approach and improving staff understanding of their behaviour by key informants. PMID- 12887631 TI - The mental health of women with suspected breast cancer: the relationship between social support, anxiety, coping and defence in maintaining mental health. AB - Relationships between anxiety, social support, coping, and defence, in connection to mental health, were studied among patients with suspected breast cancer, awaiting diagnosis. Data were collected by questionnaires from 117 women, 25-76 years of age (mean: 53.6 years) who had undergone breast biopsy. Instruments used were: the Social Provisions Scale (SPS); the state scale of State-Trait Anxiety Scale (STAI); and CODE [based on the Utrecht Coping List (UCL) and Defence Mechanisms Inventory (DMI)]. The results showed that patients reported elevated levels of anxiety and high levels of social support. Yet, anxiety was strongest and negatively related to 'instrumental coping', followed by 'cognitive defence'. 'Defensive hostility' was unrelated to anxiety. Unexpectedly, 'emotion-focused coping' and social provisions were unrelated to anxiety. Social provisions were somewhat related to 'instrumental coping', but sparsely related to 'emotion focused coping', unrelated to 'cognitive defence' and partly negatively related to 'defensive hostility'. Hence, social support and 'emotion-focused coping' did not in themselves repress anxiety. 'Instrumental coping' did, even in a situation where nothing could be done. Social support is suggested to be the product of an 'instrumental coping style', not necessarily contributing to it. Clinical consequences for professional information and support to patients with different coping styles are suggested. PMID- 12887632 TI - A trial of an anti-absconding intervention in acute psychiatric wards. AB - Absconding from acute psychiatric wards is a significant clinical problem that can place patients and others at risk, as well as being burdensome and anxiety provoking for staff. Previous studies have not convincingly demonstrated the best way to minimize the frequency of absconding. The aim of this trial was to evaluate the impact of an intervention to reduce absconding by patients from partially locked acute psychiatric wards. Five acute psychiatric wards in one hospital were entered into a stepped, before-and-after controlled trial. Following 3 months at baseline, nursing staff on the wards were trained in the intervention and monitored in its execution for the next 3 months. Absconding and violent incidents were recorded by nursing staff through shift reports and validated against officially collected forms. Absconding reduced by 25% overall during the intervention period, a fall which was statistically significant. Three out of the five wards implemented the intervention effectively and two of these achieved decreases in their absconding. The other two wards were not able to consistently implement the intervention, and their absconding rates remained unchanged. The findings support the efficacy of the intervention in reducing absconding. Further research is now required to replicate these findings, and to confirm that any reductions are maintained over time. PMID- 12887633 TI - Working with the psychological effects of trauma: consequences for mental health care workers--a literature review. AB - This literature review explores how interacting with seriously traumatized people has the potential to affect health-care workers. The review begins with an introduction to post-traumatic stress disorder as being one of the possible negative consequences of exposure to traumatic events. The report proceeds with examining the concepts of vicarious traumatization, secondary traumatic stress, traumatic countertransference, burnout and compassion fatigue, as potential adverse consequences for workers who strive to help people who are traumatized. The differences between these concepts are also discussed. The notion of compassion satisfaction is examined as findings have demonstrated that it is a protective factor which can be used as a buffer to prevent the aforementioned concepts. Conversely, findings have shown that a history of previous stressful life events in helpers is a potential risk factor. The review concludes with an overview of the concepts considered, but cautions against generalization of the findings owing to the dearth of longitudinal studies into the issues raised and also the lack of investigation into the many different types of trauma. PMID- 12887634 TI - Views of nursing staff on the use of physical restraint. AB - A postal questionnaire survey was employed in regional secure and psychiatric intensive care units in England and Wales, in respect of mental health nurses' training in the use of physical restraint. The nurses' views were sought relating to their last experience of implementing the procedure. Whilst most nurses (n = 259, 96.3%) reported positive outcomes in so far that the incident was brought under control, the views of the aftereffects of the procedure were of concern and ambivalence. The literature suggests that service users did not necessarily hold the same positive views. A range of alternatives, which were consistent with the literature, was made by staff to improve intervention in the management of violence. Negative aspects relating to the use of physical restraint were also highlighted. They included procedural, injury, clinical and management issues. Some respondents also expressed concerns about the negative attitudes of their colleagues. The findings of this aspect of the survey highlights that the therapeutic value of physical restraint can only be achieved with appropriate monitoring and with emphasis on psychological intervention in the prevention and management of violence. PMID- 12887635 TI - Partnership in coping: an Australian system of mental health nursing. AB - The paper identifies shortcomings of approaches used in organizing and delivering mental health nursing. It provides a rationale for the development of a new system of mental health nursing namely the Partnership in Coping system (PinC). This system has been developed by service users, clinical mental health nurses and an academic mental health nurse. Currently, it is undergoing trials in Western Australia. The PinC focuses on the strengths of clients. It uses the positive aspects of mental health nursing namely its holistic perspective, the length and nature of the informal contacts, the 'ordinariness' of relationships with clients and the nurse's knowledge of the clients' social and physical environments. It is a versatile system that can also be used across different mental health settings (community and inpatient facilities). The paper describes the philosophy behind the system, the concept of coping, the nature of the partnership between the client and nurse and their respective roles. PMID- 12887636 TI - CARE: A framework for responding therapeutically to the client who self-harms. AB - The CARE framework is a relevant, flexible and practical tool that can be used to map out a plan of care for helping the client who deliberately self-harms. A clear framework for care is useful for clinicians and consumers because ideally it provides a standard, holistic and considered approach for what is a complex health care issue. This paper recounts a story from clinical experience in which the CARE framework was applied. In this encounter, the nurse was helped to move beyond the limitations of the medical model towards an approach which proved satisfying for the health professional, and awakening for the client. PMID- 12887637 TI - Patients' conceptions of how health processes are promoted in mental health nursing. A qualitative study. AB - The most important goal of nursing care is to promote the subjective experience of health. The health promoting efforts of mental health care nurses must be aimed at creating encounters where the patient will be confirmed both existentially and as an individual worthy of dignity. The patient in mental health care is often viewed by the nurse as nothing more than a passive recipient of care and the belief in the patient's potential is minimal. This can lead to a situation where the patient loses control in the caring situation and feels unable to improve his/her health, which conflicts with the goal of the nursing care. The aim of the study was to describe patients' conceptions of how health processes are promoted in mental health nursing. Twelve patients with experience of mental health nursing were interviewed, and the data material was analysed using a phenomenographic approach. The results show 13 different conceptions of the phenomenon, and these were summarized into four descriptive categories: interaction, attention, development and dignity. The conceptions described show that the patients need to be treated as equals and that the nurse must trust the patient's ability to make decisions and to promote his/her health process. It is suggested that mental health nursing should be built on humanistic science and its view that every individual has the ability to grow and develop. This view is one of the most important preconditions for the promotion of health processes in mental health nursing. PMID- 12887638 TI - Coping with conflict and confusing agendas in multidisciplinary community mental health teams. AB - This paper arises from a study, conducted in the final year of general practitioner fundholding in Britain, which was designed to explore the factors influencing client allocation within six community mental health teams in the north of England. It describes the strategies employed to manage the problems created by the disparity between the stated and agreed purpose of the teams and the actual pattern of referrals. A number of strategies were identified in the six teams to manage this fundamental conflict--isolation, homogenization, fraternization, negotiation and manipulation. A sixth strategy was designed to ameliorate the loss of professional identity experienced by those who were operating in a 'foreign' milieu--that of demarcation. PMID- 12887639 TI - Caring for people in the 'virtual ward': the practical ramifications for acute nursing work. AB - This paper discusses an aspect of data analysis arising from an ethnographic study of acute mental health nursing conducted in an inner city psychiatric unit. The data were collected in the form of field notes. These were made during and following episodes of participant observation on one acute ward and on a psychiatric intensive care unit over a period of 20 months. Acute wards have been subject to continuing criticism as both sites of care and treatment for mentally ill people and of nursing work. However, the practical operations of this social world remain largely unexamined. It is argued that without this understanding, mundane work methods will always be regarded as an impediment to work that is regarded as 'therapeutic' and therefore, of importance. The focus will be on phenomena named the 'virtual ward'. The research has demonstrated that the boundaries of nurses' responsibility reach far beyond the ward's spatial environment, both physically and communicatively. The complexity of the work that takes place within the virtual ward and the skills required for its successful completion are taken for granted, but are also a practical source of frustration. Anonymized examples from the practice site studied are used to illuminate the discussion. Working within the virtual ward is part of the messy reality of nurses' work. It is argued that understanding and respecting this reality is necessary if we are to be seriously and consistently ambitious about practice development. PMID- 12887640 TI - Case management models and the care programme approach: how to make the CPA effective and credible. AB - The care programme approach (CPA), a form of case management, is a key mental health policy in England. Yet after over 10 years, it remains poorly and unevenly implemented with few benefits for service users, carers or mental health staff. This paper reviews the wider literature on case management and identifies and considers the principal models that might have informed the development of the CPA. After discussing the evidence for each of the clinical, strengths, intensive and assertive case management models, the paper identifies the key components that appear to be central to effective case management across these models. These components are then considered in relation to the CPA. It is argued that the CPA has been undermined by a failure to incorporate and build on certain important features of the major models of case management. The paper concludes by suggesting the key developments required to make the CPA more effective and to underpin the policy with a unifying philosophy while endorsing it with much needed credibility among both clinicians and service users. PMID- 12887642 TI - Practice development in psychological interventions: mental health nurse involvement in the Conversational Model of psychotherapy. AB - This paper describes a mental health nurse led practice development initiative in psychotherapy. Four mental health nurses have been trained to deliver the Conversational Model of psychotherapy (also known as psychodynamic-interpersonal (PI) psychotherapy) a non-cognitive behavioural therapy (CBT) with a robust evidence base. We report on the robust range of both processes and outcome measures being used to evaluate this initiative. We conclude that good quality evidence-based practice requires careful planning and preparation, adequate financial resources from Trusts, as well as commitment and motivation from the staff expected to be involved in such initiatives. PMID- 12887643 TI - The Promoting Engagement Project: Why is it that early interventions do not reach the children who need them most? PMID- 12887646 TI - Conference report. User involvement in curriculum and nursing practice development, the 4th Metal Health Nursing Conference, incorporating the 2nd Marion Beeforth lecture, University of Brighton, 16 April 2003. PMID- 12887647 TI - Melatonin binding sites in estrogen receptor-positive cells derived from human endometrial cancer. AB - Our previous work showed that melatonin (N-acetyl-5-methoxytryptamine) inhibits proliferation of the human endometrial cancer cell line, Ishikawa, which is estrogen receptor-positive. The aim of the present study was to determine whether Ishikawa cells possess membrane melatonin receptors. Binding of the radioligand 2 [125I]-iodomelatonin to membrane preparations obtained from Ishikawa cells was detectable, saturable and stable. Scatchard analysis revealed that the dissociation constant (Kd) of the binding sites was 179.0 pm (similar to that of the MT2 [Mel1b] melatonin receptor subtype), and that the concentration (Bmax) of the binding sites was 12.9 fmol/mg protein. Luzindole, a selective MT2 melatonin receptor antagonist, significantly suppressed binding of 2-[125I]-iodomelatonin at all concentrations tested (10(-8) to 10(-4) m). These results suggest that the MT2 melatonin receptor subtype is present in the membranes of Ishikawa cells, and that the antiproliferative effect of melatonin on Ishikawa cells is mediated via the MT2 receptor. This may have implications for the use of melatonin in endometrial cancer therapy. PMID- 12887648 TI - Protective effects of melatonin against myocardial injury induced by isoproterenol in rats. AB - This study was performed to determine whether melatonin could have a protective effect against myocardial injury (MI) induced by isoproterenol (ISO) in rats. Twenty-four rats were divided into three treatment groups: (1) control (n = 8): saline solution. (2) ISO (n = 8): ISO only. (3) melatonin + ISO (n = 8). Melatonin (10 mg/kg/day, i.p.) was administered 30 min before the initiation of ISO (150 mg/kg/day, s.c.). Drugs and saline were given at 14:00 hr for two consecutive days. At the end of the second day, blood samples were taken from the abdominal aorta shortly after the rats were anesthetized for the purpose of measuring cardiac troponins T (cTnT) and I (cTnI); hearts were removed, preserved and examined microscopically. Additionally, based on the histological changes in myocardial tissue, the rats were divided into three groups: no change, mild changes and moderate and/or marked changes. The mean cTnT and cTnI values were significantly increased in ISO group compared with control group [(1.29 +/- 0.22 ng/mL versus 0.46 +/- 0.07 ng/mL, P < 0.0001) and (0.56 +/- 0.11 ng/mL versus 0.21 +/- 0.01 ng/mL, P < 0.001)], respectively, and were significantly reduced in the ISO + melatonin group (0.65 +/- 0.06 ng/mL for cTnT and 0.25 +/- 0.01 ng/mL for cTnI) compared with the ISO only group (P < 0.01), respectively. cTnT and cTnI values were significantly increased in rats with moderate and/or marked cardiac changes compared with hearts where there were mild changes and no change (P < 0.05). ISO + melatonin group showed less histological changes than the ISO group (P < 0.01). In conclusion, this study revealed a protective effect of melatonin against ISO-induced MI in rats, and its potential clinical application in the treatment of MI. PMID- 12887649 TI - Orally administered melatonin reduces oxidative stress and proinflammatory cytokines induced by amyloid-beta peptide in rat brain: a comparative, in vivo study versus vitamin C and E. AB - To determine the efficacy of antioxidants in reducing amyloid-beta-induced oxidative stress, and the neuroinflammatory response in the central nervous system (CNS) in vivo, three injections of fibrillar amyloid-beta (fAbeta) or artificial cerebrospinal fluid (aCSF) into the CA1 region of the hippocampus of the rat were made. Concomitantly, one of the three free radical scavengers, i.e. melatonin, vitamin C, or vitamin E was also administered. Besides being a free radical scavenger, melatonin also has immunomodulatory functions. Antioxidant treatment reduced significantly oxidative stress and pro-inflammatory cytokines. There were no marked differences between melatonin, vitamin C, and vitamin E regarding their capacity to reduce nitrites and lipoperoxides. However, melatonin exhibited a superior capacity to reduce the pro-inflammatory response induced by fAbeta. PMID- 12887650 TI - Amikacin-induced acute renal injury in rats: protective role of melatonin. AB - It is well established that some agents such as aminoglycosides generate free oxygen radicals, leading to an increased oxireductase production, which in turn increases tissue toxicity. The aim of this study is to test whether melatonin, the chief secretory product of the pineal gland and a highly effective antioxidant and free radical scavenger, reduces the nephrotoxicity caused by amikacin (AK). Herein, we investigated the physiologic and pharmacological role of melatonin in influencing AK-induced nephrotoxicity. For this, pinealectomized (Px) and sham operated (non-Px) rats were used. Both AK and melatonin were administered to all groups. We investigated the effects of melatonin on AK induced changes in levels of malondialdehyde (MDA), a lipid peroxidation product, glutathione (GSH), an antioxidant whose levels are influenced by oxidative stress, and blood urea nitrogen (BUN) and serum creatine (Cr) levels. Morphologic changes in the kidney were also examined by using light microscopy. MDA levels were found to be higher in Px than in non-Px AK-treated animals. Melatonin administration to Px rats reduced MDA levels. In relative to non-Px rats, Px animals treated with AK had significantly lower GSH concentrations while melatonin administration elevated GSH levels in the kidney; however, this stimulatory effect of melatonin was not observed in non-Px AK-treated rats. Treatment with AK alone resulted in significantly higher plasma Cr and BUN levels. Repeated administration of melatonin prevented the AK-induced elevation of plasma Cr and BUN levels. Morphologic damage to renal tubules as a result of AK was more severe in the renal cortex than in the medulla. The damage to the kidney induced by AK was reversed by melatonin in the Px rats. In conclusion, these results show that physiologic melatonin concentrations are important in reducing AK-induced renal damage, while pharmacologic concentrations of melatonin did not add to the beneficial effect. PMID- 12887651 TI - Temporal integration of melatonin infusion duration: signal averaging versus frequency dependence. AB - Day length affects somatic and reproductive physiology of Siberian hamsters via regulation of the duration of nocturnal pineal melatonin secretion. Nightly 'long' (e.g. 12 hr) or 'short' (e.g. 6 hr) melatonin signals inhibit or stimulate gonadal growth, respectively. When long and short signals are presented in combination, however, neuroendocrine mechanisms exhibit a frequency-dependent response, stimulating gonadal growth only if short signals are presented every second night or more frequently. The present experiments further assessed formal models for the temporal integration of melatonin signals changing abruptly in duration from night to night. Photo-inhibited Siberian hamsters were housed in constant light and infused subcutaneously with various combinations of nightly short or long melatonin signals according to one of the several regimes that varied the frequency of short melatonin signal occurrence, average duration of the nightly melatonin signal, or both. Six weeks of nightly alternating short and long signals yielded different gonadal responses depending on the average melatonin signal duration. Moreover, when average melatonin signal duration was held constant between groups, gonadal stimulation was independent of the frequency of the constituent melatonin signals except when the duration of the short signal was reduced to 3 hr. Thus, neuroendocrine mechanisms do not solely categorize melatonin signals as either long or short but attend also to the duration of each component signal. In the majority (six of seven) of infusion regimes, reproductive responses to chimeric patterns of long and short melatonin signals were compatible with a simple signal-averaging mechanism. PMID- 12887652 TI - Melatonin prevents free radical formation due to the interaction between beta amyloid peptides and metal ions [Al(III), Zn(II), Cu(II), Mn(II), Fe(II)]. AB - Alzheimer's disease, among other pathological features, is characterized by an over-accumulation of amyloid-beta peptide, metal ions, and oxidative stress proteins in the brain. Amyloid-beta aggregated peptides with bound metal ions may initiate free radical generation with consequent protein and lipid oxidation, reactive oxygen species formation and eventually neuronal death. Melatonin is able to dramatically reduce the free radical formation which follows the interaction between transition metal ions and amyloid-beta. This paper reports the scavenging effect of melatonin of reactants generated by amyloid peptides in combination with some metal ions. PMID- 12887653 TI - Melatonin protects against lipid peroxidation and membrane rigidity in erythrocytes from patients undergoing cardiopulmonary bypass surgery. AB - The first aim of this study was to test whether there are changes in erythrocyte lipid peroxidation products and membrane fluidity during cardiac surgery involving cardiopulmonary bypass. Secondly, in vitro tests were performed to examine whether melatonin alters induced lipid peroxidation and reduced membrane fluidity in erythrocytes from these patients. Fifteen patients undergoing cardiac surgery involving cardiopulmonary bypass (CPB) were selected. Five blood samples were taken at different times during surgery for analysis of thiobarbituric acid reactive substances (TBARS) content and membrane fluidity of the erythrocytes. TBARS are an index of the level of lipid peroxidation. The results revealed an increase in TBARS levels and a parallel decrease in erythrocyte membrane fluidity (increased membrane rigidity) after the onset of CPB with respect to the reference sample. Likewise, in vitro induction of lipid peroxidation in the erythrocyte samples from CPB patients followed a similar pattern. The changes in TBARS levels and membrane fluidity were suppressed by pre-incubation of erythrocytes with melatonin prior to the induction (by 70 microm Fe2+ + 250 microm ascorbate) of lipid peroxidation in a concentration-dependent manner. The results constitute a persuasive argument for the use of melatonin for preventive and therapeutic purposes during CPB. PMID- 12887654 TI - Oxidative injury to the locus coeruleus of rat brain: neuroprotection by melatonin. AB - Neurodegeneration in the locus coeruleus (LC) has been documented in several central nervous system (CNS) neurodegenerative diseases. In the present study, iron-induced oxidative injury in the LC was investigated in chloral-hydrate anesthetized rats. Three days after bilateral infusion of iron in the LC, both vertical and horizontal locomotor activities were decreased. Seven days after unilateral infusion of iron, lipid peroxidation was elevated in the infused LC, and the norepinephrine content was depleted in the ipsilateral hippocampus of the brain. Furthermore, the immunohistochemical study demonstrated a reduction in tyrosine hydroxylase-positive neurons in the infused LC. The involvement of programmed cell death (apoptosis) in iron-induced oxidative injury in the LC was investigated. Forty-eight hours after iron infusion, cytosolic cytochrome c was elevated in the infused LC. Moreover, terminal deoxytransferase-mediated dUTP nick end labeling (TUNEL)-positive cells, an indicative of apoptosis, were detected in the infused LC. In an attempt to prevent oxidative injury in the LC, melatonin was systemically administered. Intraperitoneal injection of melatonin attenuated iron-induced behavioral changes in locomotor activity as well as iron induced increases in cytosolic cytochrome c and TUNEL-positive cells. Moreover, melatonin diminished iron-induced oxidative injury. At the same time, the level of glial derived neurotrophic factor (GDNF) was elevated in the LC of melatonin treated rats. Our data suggests that oxidative stress because of iron results in apoptosis in the infused LC and causes degeneration of the coeruleohippocampal noradrenergic system in the rat brain. Furthermore, melatonin, among other mechanisms, may exert its neuroprotection via up-regulation of GDNF levels in CNS. PMID- 12887655 TI - Long-term in vivo pineal microdialysis. AB - This study describes the development of a new technique for long-term measurement of daily 5-hydroxytryptamine (5-HT) and melatonin contents in the pineal gland of freely moving rats. The technique features a number of novel improvements over previous protocols. It allows visualization of the pineal gland for accurate targeting of the guide cannula, which minimizes bleeding; incurs no direct injury to the surrounding brain tissues; and causes no interference with the sympathetic innervation from the superior cervical ganglia. Robust releases of melatonin and indole precursors were continuously monitored quantitatively and reproducibly for more than 2 wk in the same animal. In addition, effects of pharmacological agents on in vivo pineal circadian rhythms can be studied reproducibly over time, and gene expression profiles can be correlated with physiological consequences in single animals. Using these approaches, it is found that beta-adrenergic activation leads to decreased release of 5-HT, and that increased cAMP signaling in vivo results in activation of N-acetyltransferase gene induction and melatonin production. These studies will enhance the understanding of signaling pathways that regulate pineal 5-HT and melatonin synthesis and secretion. PMID- 12887656 TI - Early neuropathological Alzheimer's changes in aged individuals are accompanied by decreased cerebrospinal fluid melatonin levels. AB - Neuropathology is the most reliable criterion for diagnosing Alzheimer's disease (AD). A well-established system for staging the spread of neuropathological changes in AD is available. The clinical use of a biomarker that reflects the neuropathological change occurring in brain tissue has not yet been established. Melatonin is a product that plays not only a major role in the regulation of the circadian rhythms but may also exert neuroprotective effects in AD. Melatonin levels were determined in ventricular postmortem cerebrospinal fluid (CSF) of 121 subjects. Braak staging and a modified Braak staging for cortex (MBSC) were used to evaluate the severity of AD neuropathology. The present study revealed that not only the Braak stages of AD, but also the MBSC were negatively correlated with CSF melatonin levels. By using MBSC, we now demonstrate for the first time that CSF melatonin levels were significantly decreased in the aged individuals with early neuropathological changes in the temporal cortex, where the AD process starts. Those individuals that did not have any neurofibrillary tangle (NFT) or neuritic plaque (NP) in the temporal cortex, had much higher melatonin levels (287 +/- 68 and 280 +/- 64 pg/mL, respectively) than those individuals that had a few NFTs and NPs (82 +/- 4 and 39 +/- 8 pg/mL, respectively) in the temporal cortex. These results suggest that the decrease in CSF melatonin levels may be an early event in the development of AD possibly occurring even before the clinical symptoms. PMID- 12887657 TI - Oxidation of melatonin by singlet molecular oxygen (O2(1deltag)) produces N1 acetyl-N2-formyl-5-methoxykynurenine. AB - It has been shown that melatonin exhibits antioxidant properties. Chemical structures of some of the products formed by the interaction of melatonin with reactive oxygen and nitrogen species have been elucidated. Despite some evidence that the reaction of melatonin with singlet molecular oxygen (O2(1deltag)) produces N1-acetyl-N2-formyl-5-methoxykynurenine (AFMK), it has not been fully documented. In this investigation, melatonin was oxidized by photosensitization with methylene blue or by a clean chemical source of O2(1deltag), the thermodecomposition of N,N'-di(2,3-dihydroxypropyl)-1,4-naphtalenedipropanamide (DHPNO2). The resulting product was characterized by high performance liquid chromatography, coupled to electrospray ionization mass spectrometry and also by 1H, 13C and dept135 nuclear magnetic resonance spectroscopy. An isotopically labeled DHPN18O2 was also prepared and used as a chemical source of labeled 18[O2(1deltag)] to unequivocally characterize the end product. The results uncovered by this work confirm the hypothesis that oxidation of melatonin by O2(1deltag) produces AFMK. PMID- 12887658 TI - Melatonin increases glutathione peroxidase activity and deformability of erythrocytes in septic rats. PMID- 12887659 TI - Unique pattern of urinary tract calculi in Australian Aboriginal children. AB - Young Aboriginal children in remote regions of tropical and desert Australia are at risk of developing urate stones in their upper urinary tract from an early age. These radiolucent calculi were only recognized with the availability of ultrasound diagnosis and are not associated with anatomic anomalies or abnormal uric acid production/metabolism. Although these stones appear to resolve spontaneously after the weaning period, some result in ureteric obstruction and infection which may lead to renal damage. This pattern of urolithiasis differs from the usual global urolithiasis pattern of either endemic bladder stones in young children in developing countries or predominantly calcium-based stones in upper tracts of older children and adults in affluent industrialized countries, where upper tract urate stones account for only a minority of childhood urinary tract stones. Risk factors for urate stones are low urine output and acidic urine. An association between urolithiasis and carbohydrate intolerance leading to chronic acidosis has been suggested for Aboriginal children, but existing limited evidence does not support this as a major aetiological factor. Although further studies on the epidemiology, natural history and management of these urate stones are needed, we believe the focus should be on improving the known social and environmental risk factors of remote Aboriginal children during the weaning period which contribute to the unacceptably high prevalence of failure to thrive, diarrhoeal disease, environmental enteropathy, iron deficiency and urolithiasis. PMID- 12887660 TI - Cows milk consumption in constipation and anal fissure in infants and young children. AB - OBJECTIVE: To examine daily cows milk consumption and duration of breastfeeding in infants and young children with anal fissure and constipation. METHODS: Two groups of 30 consecutive children aged between 4 months and 3 years were evaluated retrospectively. Group I comprised children with chronic constipation and anal fissure in whom surgical causes were excluded, and group II comprised normal children. The daily consumption of cows milk, duration of breastfeeding and other clinical features of the children were investigated RESULTS: The mean daily consumption of cows milk was significantly higher in group I (756 mL, range 200-1500 mL) than group II (253 mL, range 0-1000 mL) (P < 0.001). Group I children were breastfed for a significantly shorter period (5.8 months, range 0 18 months) than group II (10.1 months, range 2-24 months) (P < 0.006). The odds ratios for the two factors - children consuming more than 200 mL of cows milk per day (25 children in group I, 11 children in group II) and breastfeeding for less than 4 months (16 children in group I, 5 children in group II) - were calculated to be 8.6 (95% confidence interval [CI]: 0.23-0.74, P = 0.0005) and 5.7 (95% CI: 0.37-0.66, P = 0.007), respectively. CONCLUSIONS: Infants and young children with chronic constipation and anal fissure may consume larger amounts of cows milk than children with a normal bowel habit. Additionally, shorter duration of breastfeeding and early bottle feeding with cows milk may play a role in the development of constipation and anal fissure in infants and young children. PMID- 12887661 TI - Antimicrobial resistance in Helicobacter pylori isolates from children. AB - OBJECTIVE: To determine the resistance rate to four antimicrobial agents commonly used in the treatment of Helicobacter pylori infection in children. METHODS: Between July 1997 and January 2000, all H. pylori isolates from children undergoing gastroscopy were prospectively collected and subcultured to yield the susceptibility to four antimicrobial agents by E-test. In all, 23 isolates were tested. Demographic data, presenting symptoms, treatment regimen and clinical improvement after treatment were collected retrospectively. RESULTS: The resistance rate of H. pylori to metronidazole and clarithromycin were 43.5% and 8.7%, respectively. No H. pylori strains were resistant to amoxycillin or tetracycline. There were no statistically significant differences in age, sex, ethnicity, presenting symptoms or clinical improvement after treatment between antimicrobial-susceptible and antimicrobial-resistant groups. CONCLUSIONS: The frequent resistance of H. pylori to metronidazole and moderate resistance to clarithromycin in children are comparable with local adult data. The incidence of resistance tended to be higher in patients of non-European ethnicity, but this was not statistically significant. Given that the primary goal of therapy is eradication, and that local resistance rates are high, recommendations for H. pylori management may need to be modified to include sensitivity testing and/or determination of eradication in all patients. PMID- 12887662 TI - Acute disseminated encephalomyelitis: a review of 18 cases in childhood. AB - OBJECTIVE: Acute disseminated encephalomyelitis (ADEM) is a treatable inflammatory demyelinating disorder seen more commonly in children than in adults. It typically presents to general paediatricians, often, like encephalitis, with non-specific cerebrospinal fluid findings. The brain computerized tomography scan is usually normal, so is falsely reassuring and delays the diagnosis, which might result in considerable morbidity. The present study was initiated to report on the various modes of presentation and raise the awareness of the diagnosis of ADEM among general paediatricians. METHODS: A retrospective review of the case notes of 18 children with a diagnosis of ADEM established in a tertiary referral centre from 1995 to 2000 was undertaken with particular reference to clinical features, investigations and treatment. RESULTS: The most common presenting features were ataxia (10 cases), followed by headache (eight cases) and weakness (five cases). Magnetic resonance imaging (MRI) of the brain was needed to confirm the diagnosis in all 18 children. Treatment usually included a course of intravenous methylprednisolone followed by a tapering dose of oral prednisolone over several weeks. Although the outcome for most of the children was generally good, two relapsed after cessation of steroids and five children had ongoing disabilities. CONCLUSIONS: The investigation of choice for establishing the diagnosis of ADEM was MRI of the brain. Other investigations were seldom helpful in reaching the diagnosis. Early diagnosis and prompt treatment of ADEM will probably reduce morbidity. PMID- 12887663 TI - Clinical pathway using rapid rehydration for children with gastroenteritis. AB - OBJECTIVE: To determine in the Emergency Department (ED) the efficacy of a clinical pathway using rapid rehydration for children moderately dehydrated as a result of acute gastroenteritis. METHODS: This was a prospective study using historical controls, set in the ED of the Children's Hospital at Westmead, NSW, Australia. Subjects were aged from 6 months to 16 years presenting with vomiting and diarrhoea for <48 h who were mildly or moderately dehydrated. The intervention was a clinical pathway involving rapid rehydration using N/2 saline + 2.5% dextrose intravenously at 20 mL/kg per h for 2 h, or Gastrolyte R (Aventis Pharma, Lane Cove, NSW, Australia) via nasogastric tube at the same rate. There were 145 children in the prospective intervention group and 170 in the historical control group. The outcome measures were admission rate, percentage of patients discharged from the ED in 8 h or less, rate of re-presentations within 48 h requiring admission, and rate of procedures with intravenous cannula or nasogastric tube. RESULTS: In the moderately dehydrated children, significant reductions were observed in the admission rate and the number discharged in 8 h or less in the intervention group compared with the control group, with no significant difference in the rate of re-presentation and the rate of procedures. In the moderately dehydrated children in the intervention group, the admission rate was 29 of 52 (55.8%) compared with the controls 26 of 27 (96.3%) (P < 0.001) and the number discharged in 8 h or less was 23 of 52 (44.2%) compared with 1 of 27 (3.7%) in the controls (P < 0.01). CONCLUSION: The clinical pathway utilizing rapid rehydration in children moderately dehydrated from gastroenteritis is effective in reducing admission rates and lengths of stay in the ED. PMID- 12887664 TI - Neonatal plantar response revisited. AB - OBJECTIVE: To evaluate plantar response in the early neonatal period in normal, term, healthy newborn infants. This was a prospective study set in the postnatal ward of a tertiary care hospital. METHODS: The plantar response was elicited in 256 healthy, term, appropriate-for-gestational-age neonates during their first 7 days of life, utilizing the thumb-nail-drag method. The response was tested daily at intervals of 24 h, until the neonate was discharged. A total of 597 observations were made, and the responses were classified as extensor, flexor or equivocal. RESULTS: The overall plantar response was found to be predominantly extensor (73.8%), followed by equivocal (17.3%) and, finally, flexor (8.9%). The plantar response was bilaterally extensor in 72.5%, bilaterally equivocal in 14.2%, and bilaterally flexor in 7.7% of observations, respectively. Asymmetrical plantar response was elicited in 5.4% of observations. No difference was observed in individual categories based on age (<12 h, 12.1-24 h, 24.1-72 h, 72.1 h-7 days) and site of response (right/left foot). CONCLUSION: The plantar response in healthy, term neonates is predominantly extensor. Further, the relatively high frequency of asymmetrical and symmetrical flexor responses limits its clinical usefulness in the early neonatal period. PMID- 12887665 TI - Correlation among academic performance, recurrent abdominal pain and other factors in Year-6 urban primary-school children in Malaysia. AB - OBJECTIVE: The present study aimed to investigate the extent to which recurrent abdominal pain and other factors were associated with academic achievement among Year-6 (12 years of age) schoolchildren. METHODS: The present study was a cross sectional survey conducted from September to November 2001. Schoolchildren were recruited from primary schools that were selected randomly from a list of all primary schools in Petaling Jaya, Malaysia, using random sampling numbers. Information concerning recurrent abdominal pain, socio-economic status, life events, demographic and other details was obtained using a combination of questionnaires and interviews. Academic achievement was assessed using a score based on the Malaysian Primary School Achievement Examination. An overall score at or above the mean was taken to indicate high academic achievement while a score below the mean indicated poor academic achievement. RESULTS: A total of 1971 children were studied (958 boys and 1013 girls: 1047 Malays, 513 Chinese and 411 Indians). Of these children, 456 (23.1%) fulfilled the criteria for recurrent abdominal pain. Using the method of binary logistic regression analysis, the following factors were found to be independently associated with poor academic performance: a low socio-economic status (odds ratio (OR) 1.30; 95% confidence interval (CI) 1.25-1.35); male sex (OR 1.61; 95% CI 1.26-2.05); the death of a close relative (OR 2.22; 95% CI 1.73-2.85); the divorce or separation of parents (OR 3.05; 95% CI 1.73-5.40); the commencement of work by the mother (OR 1.34; 95% CI 1.02-1.76); hospitalization of the child in the 12 months prior to the study (OR 1.83; 95% CI 1.12-3.01); lack of health-care consultation (OR 1.80; 95% CI 1.36-2.36); missing breakfast (OR 1.47; 95% CI 1.07-2.02); and lack of kindergarten education (OR 1.35; 95% CI 1.04-1.75). CONCLUSIONS: Many factors, such as socio-economic status and recent life events, were associated with poor academic performance. Recurrent abdominal pain did not correlate directly to academic performance. Stress may be a means by which various factors cause children to struggle academically. PMID- 12887666 TI - Maternal phenylketonuria in Western Australia: pregnancy outcomes and developmental outcomes in offspring. AB - OBJECTIVE: To examine the outcomes of phenylketonuric (PKU) pregnancies in Western Australia including birth characteristics and cognitive and behavioural outcomes in offspring. METHODS: A cross-sectional study of women and their offspring who were identified from the Western Australian Maternal PKU Program (WAMPKUP) from 1991 to 2000 was carried out. Cognitive assessments (K-BIT or Griffiths scales) were conducted on women and their children, and behavioural assessments (CBCL) were conducted on the children. RESULTS: Thirty pregnancies by nine women were registered on the WAMPKUP between 1991 and 2000. There were 16 live births, with one preterm delivery at 32 weeks. There were no congenital abnormalities. Five of the nine mothers and their nine children (aged 18 months 10 years) participated in developmental assessments. A linear relationship was shown between lower maternal IQ scores and later attainment of metabolic control in pregnancy (rs = -0.828; P = 0.01). There was significant correlation between lower offspring IQ scores and later attainment of metabolic control in pregnancy (rs = -0.734; P = 0.02). Correlation between maternal and offspring cognitive scores was not significant. Four of nine (44%) children rated in the clinical range for behavioural problems. Compared to children with no behavioural difficulties, these children had lower cognitive abilities (P = 0.05) and maternal metabolic control during pregnancy was poor (P = 0.05). CONCLUSIONS: Poor metabolic control in pregnancy is associated with poorer cognitive outcomes and increased behavioural difficulties in offspring of mothers with PKU. The results have implications for the implementation of appropriate dietary measures before conception in PKU pregnancies, and indicate a need for the establishment of multidisciplinary teams to follow up individuals with PKU to communicate the importance of pregnancy planning, to manage PKU pregnancies, and to follow up the offspring. PMID- 12887667 TI - Audit of a paediatric directed donation programme. AB - OBJECTIVE: A paediatric directed donation programme (DDP) was instituted by Women's and Children's Health in conjunction with the Australian Red Cross Blood Service, Melbourne, Victoria, Australia, in response to public demand following a case of transfusion-transmitted HIV. This audit assesses the first 18 months of the programme. METHODS: Retrospective analysis, from February 2000 to July 2001, examining the number of units of blood requested, donated, and transfused, as well as the use of allogeneic (non-directed) blood. RESULTS: The DDP received 125 referrals. Most (78%) were for elective surgery. Of the 89 eligible children, 76% (68) had blood donated for them by an ABO/Rhesus-compatible parent, 81% of whom were first-time blood-donors. No donor tested positive for infectious markers. In total, 221 units of blood were requested and 116 units were collected. Non collection was mainly a result of parent-child ABO incompatibility or medical ineligibility of the proposed donor. Of the children for whom blood was collected, 28 (41%) received no transfusion and eight (12%) received non-directed components in addition to DDP blood; thus, 32 (47%) received solely the blood from their directed donor. Of the units collected, 53 (46%) were transfused and 63 (54%) were discarded. CONCLUSIONS: While the paediatric DDP serves a community need, the programme has a high wastage rate, is time-consuming, labour-intensive and an expensive alternative when compared with the provision of non-directed volunteer blood. In continuing the programme, appropriateness of referral needs to be refined to reduce wastage rates. PMID- 12887668 TI - Barriers to immunization: attitudes of general practitioners to varicella, the disease and its vaccine. AB - OBJECTIVES: To investigate the attitudes of general practitioners (GPs) to varicella disease and varicella vaccine. METHODS: A cross-sectional questionnaire was mailed to GPs in Fairfield (located in the south-western suburbs of Sydney, NSW, Australia) to identify attitudes about varicella vaccine, previous experience with varicella disease and the likelihood of prescribing varicella vaccine. RESULTS: Of 239 questionnaires issued, 160 were returned (67%). The majority (72%) of respondents agreed that varicella vaccine should become part of the immunization schedule. However 12% of GPs did not support vaccination for varicella as they considered it to be a benign self-limiting disease. Respondents who had experience with varicella complications were significantly more likely to recommend universal vaccination (OR 3.36; 95% confidence interval (CI) 1.38-8.19) whereas those respondents who were concerned about side effects of the vaccine were less likely to recommend universal vaccination (OR 0.31; CI 0.15-0.63). CONCLUSIONS: The majority of GPs are receptive to varicella vaccination becoming part of the immunization schedule. Experience with varicella complications is associated with recommending universal varicella vaccination. General practitioners in this cohort do not consider varicella to be a benign disease, but they are concerned about possible unknown side effects of the vaccine. Public health measures for introducing universal vaccination need to address these concerns. PMID- 12887669 TI - EpiPen epidemic: suggestions for rational prescribing in childhood food allergy. AB - There has been a marked increase in community concerns of the risk of food induced anaphylaxis in children and a consequent increase in the provision of the self or carer injectable epinephrine (EpiPen) (CSL Ltd, Parkville, Victoria, Australia)). The Australian use of EpiPens in children under 10 years has increased by 300% over 5 years with a crude rate of EpiPen provision of 1 per 544 Australian children aged under 10 years. However, the risk of a fatal reaction to food, particularly in preschool children, is remote (in Australia, an estimated one fatality in 30 years in the under 5-year-old population and two deaths in 10 years in the entire child population). It is therefore important to provide a perspective on the risk of death from food induced anaphylactic to parents and carers in view of the anxiety generated on this issue. The indications for provision of an EpiPen to children are not well defined. Six risk factors, which can be considered in evaluating the risk of a life-threatening reaction (age over 5 years; a history of respiratory tract involvement with the initial or subsequent reactions; a history of asthma requiring preventer medication; peanut or tree nut sensitivity; reactions induced by traces or small amounts of allergen; a strongly positive skin prick test) are proposed. It is suggested that the greater the number that are positive, the lower the threshold for provision of an EpiPen. In addition, instruction in EpiPen administration and the provision of both a clear and simple anaphylaxis action plan and a rational perspective on the remote risk of death is just as important as the provision of the device itself. PMID- 12887670 TI - EpiPen epidemic or good clinical practice? PMID- 12887671 TI - Benign acute childhood myositis as a cause of failure to weight bear. PMID- 12887672 TI - Carbohydrate intolerance and kidney stones in children in the Goldfields. AB - Renal stones have been reported as a common finding in Australian Aboriginal children. The stones are predominantly urate in composition. We report on five children with nephrolithiasis from the Goldfields region of Western Australia. All were diagnosed when under 5 years of age, the majority being under 3 years. All five children also had lactose intolerance, and we postulate that carbohydrate malabsorption, together with the ensuing chronic diarrhoea and intraluminal breakdown of sugars by enteric bacteria may result in a situation of chronic metabolic acidosis. Chronic metabolic acidosis can lead to protein catabolism, increased urate excretion and the formation of renal stones. Carbohydrate intolerance may be an aetiological factor in the development of renal stones and possibly chronic renal disease, particularly in Aboriginal Australians. Renal disease represents one of the most significant factors affecting the health of Australian Aboriginal people. The incidence of end stage renal failure in this population exceeds that of non-Aboriginals by a factor of 13:1, and this disproportionate figure is increasing. It is likely that chronic renal damage is multifactorial; however, it is probable that at least some aetiological factors have their onset during childhood. PMID- 12887673 TI - Parenteral nutrition solution retrieved by lumbar puncture following left saphenous vein catheterization. AB - Two extremely preterm infants had silastic lines inserted via the left great saphenous vein for intravenous feeding. Clinical deterioration approximately 10 days later led to lumbar puncture as part of a sepsis screen. Parenteral nutrition solution was recovered in the cerebrospinal fluid and heralded potentially life-threatening catheter malposition. The cases illustrate another percutaneous central line-associated morbidity, which is avoidable by careful verification of line position. All previously reported cases of this unusual complication are reviewed and we describe its likely mechanism. We postulate that the phenomenon is a complication peculiar to percutaneous catheterization of the left lower limb. PMID- 12887674 TI - Red eyes as the initial presentation of systemic meningococcal infection. AB - The present paper is a report of a 14-month-old boy who presented with fever, coryzal symptoms and red eyes. The patient developed a generalized tonic clonic convulsion on day 2 of his illness. Ophthalmological assessment demonstrated bilateral hypopyon and vitreous opacity resulting from endophthalmitis. Cerebrospinal fluid was positive for Neisseria meningitidis (A, C, Y, W 135) by latex agglutination. He was treated with high dose intravenous cefotaxime and intravitreal ceftazidime. He made good recovery and his vision was preserved. In view of the potential morbidity and mortality associated with systemic meningococcal infection, the presence of red eye and hypopyon provides important diagnostic clues indicating the need to investigate beyond superficial conjunctivitis. It should prompt the clinician to recognize endopthalmitis early and accurately diagnose this serious disease. PMID- 12887675 TI - Velcroholism. PMID- 12887676 TI - Ivermectin for scabies. PMID- 12887677 TI - Metabotropic glutamate type 1alpha receptor localizes in low-density caveolin rich plasma membrane fractions. AB - Recent evidence suggest that many G protein-coupled receptors (GPCR) and signalling molecules localize in microdomains of the plasma membrane. In this study, flotation gradient analysis in the absence of detergents demonstrated the presence of the metabotropic glutamate receptor type 1alpha (mGlu1alpha) in low density caveolin-enriched membrane fractions (CEMF) in permanently transfected BHK cells. BHK-1alpha cells exhibit a similar pattern of staining for caveolin-1 and caveolin-2, and these two proteins show a high degree of co-localization with mGlu1alpha receptor as demonstrated by immunogold and confocal laser microscopy. The presence of mGlu1alpha in CEMF was also demonstrated by co immunoprecipitation of mGlu1alpha receptor using antibodies against caveolin proteins. Activation of the mGlu1alpha receptor by agonist increased extracellular signal-regulated kinases phosphorylation in CEMF and not in high density membrane fractions (HDMF), suggesting that mGlu1alpha receptor-mediated signal transduction could occur in caveolae-like domains. Overall, these results clearly show a molecular and functional association of mGlu1alpha receptor with caveolins. PMID- 12887678 TI - Mitochondrial apoptotic cell death and moderate superoxide generation upon selective activation of non-desensitizing AMPA receptors in hippocampal cultures. AB - In the present work we investigated the effect of selective stimulation of non desensitizing alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA) receptors in the intracellular processes leading to hippocampal neuronal death and production of reactive oxygen species (ROS). Activation of AMPA receptors in the presence of cyclothiazide (CYZ), a blocker of AMPA receptor desensitization, resulted in the death of approximately 25% of neurones, which was prevented by 2,3-dihydroxy-6-nitro-7-sulphamoyl-benzo(f)quinoxaline (NBQX), an AMPA-preferring receptor antagonist. (+)-5-Methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10 imine hydrogen maleate (MK-801) protected the neurones from necrotic death induced by AMPA or NMDA receptor activation. Neurodegeneration caused by selective activation of non-desensitizing AMPA receptors, in the presence of AMPA, CYZ and MK-801, significantly decreased the number of Co2+-positive neurones, used as a cytochemical marker of Ca2+-permeable AMPA receptors, but maintained intracellular ATP/ADP. The AMPA-mediated apoptotic cell death involved mitochondrial cytochrome c release and the activation of caspases-1 and -3, which was prevented by NBQX. Interestingly, although selective activation of AMPA receptors was not associated with production of intracellular peroxides, a moderate increase in superoxide production was observed upon exposure to antimycin A (AA). Furthermore, increased activity of Mn- superoxide dismutase (SOD) was observed on selective activation of non-desensitizing AMPA receptors. Taken together, these data make important contributions to the elucidation of the downstream pathways activated in AMPA receptor-mediated excitotoxicity in cultured rat hippocampal neurones. PMID- 12887679 TI - CREB is required for acquisition of ischemic tolerance in gerbil hippocampal CA1 region. AB - Ischemic tolerance is well known as a neuroprotective effect of pre-conditioning ischemia against delayed neuronal death, however, the mechanism or mechanisms underlying this effect are not fully understood. We investigated the relationship between CREB and ischemic tolerance in gerbil hippocampal CA1 neurons using CRE decoy oligonucleotide. Sublethal ischemia led to an increase in the level of CREB phosphorylation in CA1 regions while lethal ischemia did not. Experiments with NG108-15 cells showed that adding CRE decoy oligonucleotide to culture media significantly inhibited the cell growth rate. The administration of CRE decoy oligonucleotide into gerbil cerebral ventricle decreased CREB-DNA binding activity to 38% of the control. Pre-treatment with CRE decoy oligonucleotide 24 h before the induction of ischemic tolerance decreased CA1 neuronal cell survival to 21% of the control. The present findings suggest that a CREB-mediated transcription system is necessary for the induction of ischemic tolerance. PMID- 12887680 TI - Dynorphin A(1-17) biotransformation in striatum of freely moving rats using microdialysis and matrix-assisted laser desorption/ionization mass spectrometry. AB - The biotransformation of the opioid peptide dynorphin A(1-17) was investigated in striatum of freely moving Fischer rats, by direct infusion of this peptide, followed by recovery of the resulting biotransformation products via microdialysis and identification using matrix-assisted laser desorption/ionization mass spectrometry. The observed peptides are consistent with enzymatic cleavage at the Arg7-Ile8 position of dynorphin A(1-17), followed by terminal degradation of the resulting dynorphin A(1-7) and dynorphin A(8-17) peptides. Unexpectedly, novel post-translational modifications were found on C terminal fragments of dynorphin A(1-17). Using tandem mass spectrometry, a covalent modification of mass 172 Da, the nature of which is not understood, was found on the tryptophan residue of C-terminal fragments (Trp14). Additional modifications, of mass 42 and 113 Da, were also found on the N-terminus (Ile8 or Pro10) of these same C-terminal fragments. The role of these modifications of C terminal fragments has not yet been characterized. PMID- 12887681 TI - Characterization of the N-acetylaspartate biosynthetic enzyme from rat brain. AB - Aspartate N-acetyltransferase (Asp-NAT; EC 2.3.1.17) activity was found in highly purified intact mitochondria prepared by Percoll gradient centrifugation as well as in the three subfractions obtained after the sucrose density gradient centrifugation of Percoll purified mitochondria; citrate synthase was used as a marker enzyme for mitochondria. The proportion of recoverable activities of Asp NAT and citrate synthase were comparable in mitochondrial and synaptosomal fractions but not in the fraction containing myelin. Asp-NAT was solubilized from the pellet of the rat brain homogenate (26 000 g for 1 h) for the recovery of maximum activity and partially purified using three protein separation methods: DEAE anion exchange chromatography, continuous elution native gel electrophoresis and size-exclusion high performance liquid chromatography. Asp-NAT activity and the optical density pattern of the eluted protein from size-exclusion column indicated a single large protein (approximately 670 kDa), which on sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed at least 10 bands indicative of an enzyme complex. This seemingly multi-subunit complex Asp-NAT was stable towards ionic perturbations but vulnerable to hydrophobic perturbation; almost 95% of activity was lost after 10 mm 3-[(3-cholamidopropyl)dimethylammonia]-1 propanesulfonate (CHAPS) treatment followed by size-exclusion chromatography. Asp NAT showed an order of magnitude difference in Km between l-aspartate (l-Asp, approximately 0.5 mm) and acetyl CoA (approximately 0.05 mm). Asp-NAT showed high specificity towards l-Asp with 3% or less activity towards l-Glu, l-Asn, l-Gln and Asp-Glu. A model on the integral involvement of NAA synthesis in the energetics of neuronal mitochondria is proposed. PMID- 12887682 TI - Distinct cleavage patterns of normal and pathologic forms of alpha-synuclein by calpain I in vitro. AB - Parkinson's disease (PD) is characterized by fibrillary neuronal inclusions called Lewy bodies (LBs) consisting largely of alpha-synuclein (alpha-syn), the protein mutated in some patients with familial PD. The mechanisms of alpha-syn fibrillization and LB formation are unknown, but may involve aberrant degradation or turnover. We examined the ability of calpain I to cleave alpha-syn in vitro. Calpain I cleaved wild-type alpha-syn predominantly after amino acid 57 and within the non-amyloid component (NAC) region. In contrast, calpain I cleaved fibrillized alpha-syn primarily in the region of amino acid 120 to generate fragments like those that increase susceptibility to dopamine toxicity and oxidative stress. Further, while calpain I cleaved wild-type alpha-syn after amino acid 57, this did not occur in mutant A53T alpha-syn. This paucity of proteolysis could increase the stability of A53T alpha-syn, suggesting that calpain I might protect cells from forming LBs by specific cleavages of soluble wild-type alpha-syn. However, once alpha-syn has polymerized into fibrils, calpain I may contribute to toxicity of these forms of alpha-syn by cleaving at aberrant sites within the C-terminal region. Elucidating the role of calpain I in the proteolytic processing of alpha-syn in normal and diseased brains may clarify mechanisms of neurodegenerative alpha-synucleinopathies. PMID- 12887683 TI - Progesterone and its metabolites increase myelin basic protein expression in organotypic slice cultures of rat cerebellum. AB - We have previously shown that progesterone (PROG) is synthesized by Schwann cells and promotes myelin formation in the peripheral nervous system (PNS). We now report that this neurosteroid also stimulates myelination in organotypic slice cultures of 7-day-old (P7) rat and mouse cerebellum. Myelination was evaluated by immunofluorescence analysis of the myelin basic protein (MBP). After 7 days in culture (7DIV), we found that adding PROG (2(-5) x 10(-5) M) to the culture medium caused a fourfold increase in MBP expression when compared to control slices. The effect of PROG on MBP expression involves the classical intracellular PROG receptor (PR): the selective PR agonist R5020 significantly increased MBP expression and the PR antagonist mifepristone (RU486) completely abolished the effect of PROG on this MBP expression. Moreover, treatment of P7-cerebellar slice cultures from PR knockout (PRKO) mice with PROG had no significant effect on MBP expression. PROG was metabolized in the cerebellar slices to 5alpha dihydroprogesterone (5alpha-DHP) and to the GABAA receptor-active metabolite 3alpha,5alpha-tetrahydroprogesterone (3alpha,5alpha-THP, allopregnanolone). The 5alpha-reductase inhibitor L685-273 partially inhibited the effect of PROG, and 3alpha,5alpha-THP (2(-5) x 10(-5) M) significantly stimulated the MBP expression, although to a lesser extent than PROG. The increase in MBP expression by 3alpha,5alpha-THP involved GABAA receptors, as it could be inhibited by the selective GABAA receptor antagonist bicuculline. These findings suggest that progestins stimulate MBP expression and consequently suggest an increase in CNS myelination via two signalling systems, the intracellular PR and membrane GABAA receptors, and they confirm a new role of GABAA receptors in myelination. PMID- 12887684 TI - Mitochondrial peroxiredoxin-3 protects hippocampal neurons from excitotoxic injury in vivo. AB - Mitochondria are involved in excitotoxic damage of nerve cells. Following the breakdown of the calcium-buffering ability of mitochondria, mitochondrial calcium overload induces reactive oxygen species (ROS) bursts that produce free radicals and open permeability transition pores, ultimately leading to neuronal cell death. In the present study, we focused on a mitochondrial antioxidant protein, peroxiredoxin-3 (Prx-3), to investigate the mechanism by which toxic properties of ROS were up-regulated in mitochondria of damaged nerve cells. Immunohistochemical analysis revealed that Prx-3 protein exists in mitochondria of rat hippocampus, whereas we found a significant decrease in Prx-3 mRNA and protein levels associated with an increase in nitrated proteins in the rat hippocampus injured by microinjection of ibotenic acid. Furthermore, in vivo adenoviral gene transfer of Prx-3 completely inhibited protein nitration and markedly reduced gliosis, a post-neuronal cell death event. Since mitochondrial Prx-3 seems to be neuroprotective against oxidative insults, our findings suggest that Prx-3 up-regulation might be a useful novel approach for the management of neurodegenerative diseases. PMID- 12887685 TI - Cloning and characterization of a Caenorhabditis elegans D2-like dopamine receptor. AB - The neurotransmitter dopamine plays an important role in the regulation of behavior in both vertebrates and invertebrates. In mammals, dopamine binds and activates two classes of dopamine receptors, D1-like and D2-like receptors. However, D2-like dopamine receptors in Caenorhabditis elegans have not yet been characterized. We have cloned a cDNA encoding a putative C. elegans D2-like dopamine receptor. The deduced amino acid sequence of the cloned cDNA shows higher sequence similarities to vertebrate D2-like dopamine receptors than to D1 like receptors. Two splice variants that differ in the length of their predicted third intracellular loops were identified. The receptor heterologously expressed in cultured cells showed high affinity binding to [125I]iodo-lysergic acid diethylamide. Dopamine showed the highest affinity for this receptor among several amine neurotransmitters tested. Activation of the heterologously expressed receptor led to the inhibition of cyclic AMP production, confirming that this receptor has the functional property of a D2-like receptor. We have also analyzed the expression pattern of this receptor and found that the receptor is expressed in several neurons including all the dopaminergic neurons in C. elegans. PMID- 12887687 TI - Phorbol ester induces CYP2E1 in astrocytes, through a protein kinase C- and tyrosine kinase-dependent mechanism. AB - Cytochrome P450 2E1 (CYP2E1) is highly inducible in a subset of astrocytes in vivo following ischemic or mechanical injury and in vitro by lipopolysaccharide or interleukin-1beta. In the present study, phorbol-12,13-dibutyrate (PDBu) was found to induce catalytically active CYP2E1 more than fourfold in cortical glial cultures. Little induction was seen up to 12 h, and full effects only at 21-24 h of PDBu treatment. CYP2E1 expression in PDBu-treated cells was enriched in a subset of astrocytes. The protein kinase C inhibitors, staurosporine and calphostin C, and the tyrosine kinase inhibitor genistein, but not its inactive analogue daidzein, prevented the induction of CYP2E1 by PDBu. It is suggested that CYP2E1, together with interleukin-6 and ciliary neurotrophic factor, is part of a response of astrocytes to cellular stress elicited by, e.g. cerebral injury, cytokines or phorbol ester, and mediated in part through protein kinase C. PMID- 12887686 TI - Kinetic differences between the isoforms of glutamate decarboxylase: implications for the regulation of GABA synthesis. AB - Glutamate decarboxylase (GAD) exists as two isoforms, GAD65 and GAD67. GAD activity is regulated by a cycle of activation and inactivation determined by the binding and release of its co-factor, pyridoxal 5'-phosphate. Holoenzyme (GAD with bound co-factor) decarboxylates glutamate to form GABA, but it also catalyzes a slower transamination reaction that produces inactive apoGAD (without bound co-factor). Apoenzyme can reassociate with pyridoxal phosphate to form holoGAD, thus completing the cycle. Within cells, GAD65 is largely apoenzyme (approximately 93%) while GAD67 is mainly holoenzyme (approximately 72%). We found striking kinetic differences between the GAD isoforms that appear to account for this difference in co-factor saturation. The glutamate dependent conversion of holoGAD65 to apoGAD was about 15 times faster than that of holoGAD67 at saturating glutamate. Aspartate and GABA also converted holoGAD65 to apoGAD at higher rates than they did holoGAD67. Nucleoside triphosphates (such as ATP) are known to affect the activation reactions of the cycle. ATP slowed the activation of GAD65 and markedly reduced its steady-state activity, but had little affect on the activation of GAD67 or its steady-state activity. Inorganic phosphate opposed the effect of ATP; it increased the rate of apoGAD65 activation but had little effect on apoGAD67 activation. We conclude that the apo /holoenzyme cycle of inactivation and reactivation is more important in regulating the activity of GAD65 than of GAD67. PMID- 12887688 TI - Effect of middle cerebral artery occlusion on mRNA expression for the sodium coupled vitamin C transporter SVCT2 in rat brain. AB - The sodium-vitamin C co-transporter SVCT2 is primarily responsible for the accumulation of the important antioxidant ascorbate into brain cells. In vitro studies have demonstrated strong expression of this transporter in cultured astrocytes, whereas in situ hybridization analysis has so far detected SVCT2 only in neurons. In the present study, we examined the response of SVCT2 mRNA expression in the brain to focal ischemia induced for 2 h by unilateral middle cerebral artery occlusion. The mRNA expression patterns of SVCT2 and the glutamate-activated immediate early gene Arc were investigated at 2 and 22 h after ischemia. SVCT2 and Arc mRNA expression was lost in the ischemic core at both time points. In areas outside the core, Arc was strongly up-regulated, primarily at 2 h, whereas SVCT2 showed an increase at 2 and 22 h. SVCT2 expression was increased in neurons as well as in astrocytes, providing the first evidence for SVCT2 expression in astrocytes in situ. These findings underscore the importance of ascorbate as a neuroprotective agent and may have implications for therapeutic strategies. In addition, the increase of SVCT2 in astrocytes after ischemia suggests that cultured astrocytes are exposed to chronic oxidative stress. PMID- 12887689 TI - Noradrenaline induces expression of peroxisome proliferator activated receptor gamma (PPARgamma) in murine primary astrocytes and neurons. AB - Cerebral inflammatory events play an important part in the pathogenesis of Alzheimer's disease (AD). Agonists of the peroxisome proliferator-activated receptor gamma (PPARgamma), a nuclear hormone receptor that mediates anti inflammatory actions of non-steroidal anti-inflammatory drugs (NSAIDs) and thiazolidinediones, have been therefore proposed as a potential treatment of AD. Experimental evidence suggests that cortical noradrenaline (NA) depletion due to degeneration of the locus ceruleus (LC) - a pathological hallmark of AD - plays a permissive role in the development of inflammation in AD. To study a possible relationship between NA depletion and PPARgamma-mediated suppression of inflammation we investigated the influence of NA on PPARgamma expression in murine primary cortical astrocytes and neurons. Incubation of astrocytes and neurons with 100 micro m NA resulted in an increase of PPARgamma mRNA as well as PPARgamma protein levels in both cell types. These effects were blocked by the beta-adrenergic antagonist propranolol but not by the alpha-adrenergic antagonist phentolamine, suggesting that they might be mediated by beta-adrenergic receptors. Our results indicate for the first time that PPARgamma expression can be modulated by the cAMP signalling pathway, and suggest that the anti inflammatory effects of NA on brain cells may be partly mediated by increasing PPARgamma levels. Conversely, decreased NA due to LC cell death in AD may reduce endogenous PPARgamma expression and therefore potentiate neuroinflammatory processes. PMID- 12887690 TI - Regulation of expression of sulfoglucuronyl carbohydrate (HNK-1), Amphoterin and RAGE in retinoic acid-differentiated P19 embryonal carcinoma cells. AB - HNK-1 antibody reactive sulfoglucuronyl carbohydrate (SGC) and SSEA-1 antibody reactive Lewis X (Lex) epitope are expressed on several glycolipids, glycoproteins, and proteoglycans of the nervous system and have been implicated in cell-cell recognition, neurite outgrowth, and/or neuronal migration during development. Interaction of SGC with its binding protein Amphoterin and interaction of Amphoterin with a cell-signaling molecule, receptor for advance glycation end product (RAGE) have been suggested to regulate neurite outgrowth and neuronal migration. The regulation of expression of SGC, Lex, Amphoterin, and RAGE was studied in embryonal carcinoma P19 cells after treatment with retinoic acid (RA). The untreated proliferating P19 cells strongly expressed the Lex epitope, which was mostly due to Lex-glycoproteins. P19 cells, when differentiated into neuron-like cells by RA, did not express the Lex epitope, but expressed increasing levels of SGC, with time in culture. Quantitative biochemical analyses showed that in the P19 cells after RA treatment, the amount of SGC-glycoproteins increased at a significantly higher level than sulfoglucuronyl glycolipid-1 (SGGL-1). The increase in the levels of SGGL-1 was due to 16-fold upregulation in the activity of lactosylceramide: N acetylglucosaminyl-transferase (Lc3 synthase), which synthesizes the key intermediate lactotriosylceramide (Lc3Cer), for lacto- and neolacto-glycolipids. The large increase in the activity of Lc3 synthase appeared to regulate the levels of other neolacto glycolipids, such as Lc3Cer, nLc4Cer, nLc6Cer, disialosyl-nLc4Cer (LD1), and Lex-glycolipids. Strong upregulation of glucuronyl transferase and modest twofold enhancement in the activity of the glucuronyl sulfotransferase, which catalyze the final steps in the SGC synthesis, also would account for the large increase in the synthesis SGC-glycoproteins. RA also upregulated the synthesis of Amphoterin and RAGE in P19 cells. SGC, RAGE, and Amphoterin were co-localized in the RA-differentiated neurons. The initiation of neurite outgrowth along with co-ordinated upregulation of Amphoterin, RAGE, SGC glycoproteins, and SGGLs in RA-treated P19 cells support the hypothesis that these molecules are involved in the neuronal process formation. PMID- 12887691 TI - Microdialysis of dopamine interpreted with quantitative model incorporating probe implantation trauma. AB - Although microdialysis is widely used to sample endogenous and exogenous substances in vivo, interpretation of the results obtained by this technique remains controversial. The goal of the present study was to examine recent criticism of microdialysis in the specific case of dopamine (DA) measurements in the brain extracellular microenvironment. The apparent steady-state basal extracellular concentration and extraction fraction of DA were determined in anesthetized rat striatum by the concentration difference (no-net-flux) microdialysis technique. A rate constant for extracellular clearance of DA calculated from the extraction fraction was smaller than the previously determined estimate by fast-scan cyclic voltammetry for cellular uptake of DA. Because the relatively small size of the voltammetric microsensor produces little tissue damage, the discrepancy between the uptake rate constants may be a consequence of trauma from microdialysis probe implantation. The trauma layer has previously been identified by histology and proposed to distort measurements of extracellular DA levels by the no-net-flux method. To address this issue, an existing quantitative mathematical model for microdialysis was modified to incorporate a traumatized tissue layer interposed between the probe and surrounding normal tissue. The tissue layers are hypothesized to differ in their rates of neurotransmitter release and uptake. A post-implantation traumatized layer with reduced uptake and no release can reconcile the discrepancy between DA uptake measured by microdialysis and voltammetry. The model predicts that this trauma layer would cause the DA extraction fraction obtained from microdialysis in vivo calibration techniques, such as no-net-flux, to differ from the DA relative recovery and lead to an underestimation of the DA extracellular concentration in the surrounding normal tissue. PMID- 12887692 TI - Molecular modeling and mutagenesis of the ligand-binding pocket of the mGlu3 subtype of metabotropic glutamate receptor. AB - A homology model of the extracellular domain of the mGlu3 subtype of metabotropic glutamate (mGlu) receptor was generated and tested using site-directed mutagenesis, a radioligand-binding assay using the Group II selective agonist (2S,2'R,3'R)-2-(2',3'-[3H]dicarboxycyclopropyl) glycine ([3H]DCG-IV), and in a fluorescence-based functional assay in live transiently transfected human embryonic kidney cells. Ten of the 12 mGlu3 mutants (R64A, R68A, Y150A, S151A, T174A, D194A, Y222A, R277A, D301A and K389) showed either no binding or a 90% or greater loss of specific [3H]DCG-IV binding. Several analogous mutations in mGlu2 supported the results obtained with mGlu3. These results demonstrate that the binding of [3H]DCG-IV to mGlu3 is exceptionally sensitive to mutagenesis-induced perturbations. In silico docking of DCG-IV into the agonist binding pocket of mGlu3 facilitated the interpretation the mutagenesis results. Tyrosines 150 and 222, and arginine 277 show close contacts with the third carboxylic acid group in DCG-IV, which is not present in glutamate or (2S,1'S,2'S)-2 (carboxycyclopropyl)glycine (L-CCG-I). Mutation of these three amino acids to alanine resulted in a near complete loss of receptor activation by DCG-IV and retention of near wild-type affinity for L-CCG-I. It is proposed that hydrogen bonding between this carboxylate and tyrosines 150 and 222 and arginine 277 provide a partial explanation for the high affinity and Group II selectivity of DCG-IV. These findings define the essential features of the ligand-binding pocket of mGlu3 and, together with other recent studies on mGlu receptors, provide new opportunities for structure-based drug design. PMID- 12887693 TI - Functional coupling of serotonin and noradrenaline transporters. AB - Re-uptake of the neurotransmitters serotonin and noradrenaline out of the synaptic cleft is mediated by selective transporter proteins, the serotonin transporter and the noradrenaline transporter respectively. Both are integral membrane proteins that are have a high degree of homology and represent members of a larger neurotransmitter transporter superfamily. Several studies have indicated that the serotonin transporter has an an oligomeric structure. To determine whether monoamine transporters can also function in oligomeric structures in situ, we constructed a concatenate consisting of one molecule of serotonin transporter covalently linked to one molecule of noradrenaline transporter. Heterologous expression of this hybrid construct allowed us to analyse the function, i.e. transport activity, and the structure, i.e. the molecular weight of the total construct and of its single components, at the same time. We showed that serotonin-noradrenaline transporter fusion proteins are fully active and exhibit the pharmacological profile of both their individual components. These findings support the hypothesis that monoamine transporters are expressed and may function as oligomeric proteins composed of non-interacting monomers. PMID- 12887694 TI - Involvement of mitochondrial K+ release and cellular efflux in ischemic and apoptotic neuronal death. AB - We measured and manipulated intracellular potassium (K+) fluxes in cultured hippocampal neurons in an effort to understand the involvement of K+ in neuronal death under conditions of ischemia and exposure to apoptotic stimuli. Measurements of the intracellular K+ concentration using the fluorescent probe 1,3-benzenedicarboxylic acid, 4,4'-[1,4,10,13-tetraoxa-7,16-diazacyclooctadecane 7,16-diylbis(5-methoxy-6,2-benzofurandiyl)]bis-, tetrakis [(acetyloxy) methyl] ester (PBFI) revealed that exposure of neurons to cyanide (chemical hypoxia), glutamate (excitotoxic insult) or staurosporine (apoptotic stimulus) results in efflux of K+ and cell death. Treatment of neurons with 5-hydroxydecanoate (5HD), an inhibitor of mitochondrial K+ channels, reduced K+ fluxes in neurons exposed to each insult and increased the resistance of the cells to death. K+ efflux was attenuated, levels of oxyradicals were decreased, mitochondrial membrane potential was stabilized and release of cytochrome c from mitochondria was attenuated in neurons treated with 5HD. K+ was rapidly released into the cytosol from mitochondria when neurons were exposed to the K+ channel opener, diazoxide, or to the mitochondrial uncoupler, carbonyl cyanide 4(trifluoromethoxy)phenylhydrazone (FCCP), demonstrating that the intramitochondrial K+ concentration is greater than the cytosolic K+ concentration. The release of K+ from mitochondria was followed by efflux through plasma membrane K+ channels. In vivo studies showed that 5HD reduces ischemic brain damage without affecting cerebral blood flow in a mouse model of focal ischemic stroke. These findings suggest that intracellular K+ fluxes play a key role in modulating neuronal oxyradical production and cell survival under ischemic conditions, and that agents that modify K+ fluxes may have therapeutic benefit in stroke and related neurodegenerative conditions. PMID- 12887695 TI - A complex signaling cascade links the serotonin2A receptor to phospholipase A2 activation: the involvement of MAP kinases. AB - Previous studies in our laboratory have shown that in NIH3T3-5HT2A cells, 5-HT induced AA release is PLA2-coupled and independent of 5-HT2A receptor-mediated PLC activation. Although 5-HT2A receptor-mediated PLC activation is known to be Galphaq-coupled, much less is understood about 5-HT2A receptor-mediated PLA2 activation. Therefore, the studies presented here were aimed at elucidating the signal transduction pathway linking stimulation of the 5-HT2A receptor to PLA2 activation. By employing various selective inhibitors, toxins, and antagonistic peptide constructs, we propose that the 5-HT2A receptor can couple to PLA2 activation through two parallel signaling cascades. Initial experiments were designed to examine the role of pertussis toxin-sensitive G proteins, namely Galphai/o, as well as pertussis toxin-insensitive G proteins, namely Galpha12/13, in 5-HT-induced AA release. Furthermore, inactivation of both Gbetagamma heterodimers and Rho proteins resulted in decreased agonist-induced AA release, without having any effect on PLC-IP accumulation. We also demonstrated 5-HT2A receptor-mediated phosphorylation of ERK1,2 and p38. Moreover, pretreatment with selective ERK1,2 and p38 inhibitors resulted in decreased 5-HT-induced AA release. Taken together, these results suggest that the 5-HT2A receptor expressed in NIH3T3 cells can couple to PLA2 activation though a complex signaling mechanism involving both Galphai/o-associated Gbetagamma-mediated ERK1,2 activation and Galpha12/13-coupled, Rho-mediated p38 activation. PMID- 12887696 TI - The role of the RE1 element in activation of the NR1 promoter during neuronal differentiation. AB - To understand the genetic mechanism controlling the expression of the NMDA subtype of glutamate receptors during neuronal differentiation, we studied activation of the N-methyl-D-aspartate receptor subunit 1 (NR1) gene and the role of the repressor element-1 (RE1) element in NR1 promoter activation. Following neuronal differentiation of P19 embryonic carcinoma cells, the NR1 transcription rate and mRNA level were significantly increased, while the nuclear level of the repressor RE1 silencing transcription factor (REST)/neuron-restriction silencer factor (NRSF) was reduced. Nuclear REST/NRSF from undifferentiated cells formed a large complex with the NR1 RE1 element. While this complex was significantly reduced after the differentiation, REST/NRSF from differentiated cells formed a new, faster migrating complex. In transient transfections, deletion of the RE1 element increased activity of the 5.4-kb NR1 promoter sixfold in undifferentiated cells, but only induced approximately 1.4-fold increase in differentiated cells. Forced expression of REST/NRSF in differentiated cells suppressed the promoter, while forced expression of a dominant-negative REST/NRSF induced promoter activity as well as the mRNA of the NR1 gene in undifferentiated cells. In stable transfectants, the wild-type promoter showed a robust increase in activity following differentiation in a pattern similar to the NR1 mRNA increase. Conversely, the promoter lacking the RE1 element showed only a moderate increase. Our data suggest that the NR1 gene up-regulation during neuronal differentiation is controlled by its promoter activation, which is largely determined by the interaction between the RE1 element and the repressor REST/NRSF. PMID- 12887697 TI - Diabetic neuropathy: inhibitory G protein dysfunction involves PKC-dependent phosphorylation of Goalpha. AB - We examined the hypothesis that decreased inhibitory G protein function in diabetic neuropathy is associated with increased protein kinase C (PKC)-dependent phosphorylation of the Goalpha subunit. Streptozotocin-induced diabetic rats were studied between 4 and 8 weeks after onset of diabetes and compared with aged matched healthy animals as controls. Opioid-mediated inhibition of forskolin stimulated cyclic AMP was significantly less in dorsal root ganglia (DRGs) from diabetic rats compared with controls. Activation of PKC in DRGs from control rats was associated with a significant decrease in opioid-mediated inhibition of forskolin-stimulated cyclic AMP that was similar to the decrease in inhibition observed in DRGs from diabetic rats. Both basal and PKC-mediated labeling of Goalpha with 32Pi was significantly less in DRGs from diabetic rats, supporting increased endogenous PKC-dependent phosphorylation of Goalpha. Probing of immunoprecipitated Goalpha with an anti-phospho-serine/threonine specific antibody revealed a significant increase in baseline phosphorylation in diabetic DRGs. Activation of PKC produced a significant increase in phosphorylation in control DRGs but no significant increase in Goalpha in diabetic DRGs. Phosphorylation of PKC-alpha was increased, PKC-betaII was unchanged and PKC delta decreased in diabetic DRGs. These results suggest that diminished inhibitory G protein function observed in DRGs neurons from diabetic rats involves an isoform-specific PKC-dependent pathway. PMID- 12887698 TI - The structure of neuromelanin as studied by chemical degradative methods. AB - The biosynthesis, structure and function of neuromelanin (NM), the dark brown melanin-like pigment present in the substantia nigra (SN), are not well characterized, in spite of the possible involvement of NM in the etiology and pathogenesis of Parkinson's disease. NM was isolated from the SN of five non Parkinsonian human brains. NM and synthetic melanins, employed as models, were characterized by chemical analysis. Alkaline hydrogen peroxide (H2O2) oxidation of NM generated four degradation products, pyrrole-2,3-dicarboxylic acid (PDCA), pyrrole-2,3,5-tricarboxylic acid (PTCA), thiazole-4,5-dicarboxylic acid (TDCA) and thiazole-2,3,5-tricarboxylic acid (TTCA), whose ratios, especially the TTCA to PDCA ratio, indicate that NM is derived mostly from dopamine (DA) with 25% incorporation of cysteine (Cys) in the form of a benzothiazine structure. Hydriodic acid (HI) reductive hydrolysis of NM yielded 4-amino-3 hydroxyphenylethylamine (4-AHPEA) as a marker of cysteinyldopamine (CysDA) derived units. The 4-AHPEA to PDCA ratio indicates a 21% incorporation of CysDA derived units into NM. These degradative experiments also suggest that DOPA is not incorporated into NM to a significant extent (approximately 6% the level of DA). It is concluded that the TTCA to PDCA ratio is a useful indicator of CysDA derived units in NM, and NM consists mainly of DA-melanin with some contribution from CysDA-melanin. The involvement of DA and CysDA as building blocks of NM demonstrates the detoxifying role of NM synthesis, since it prevents the intraneuronal accumulation of DA and CysDA, which would cause toxic effects. PMID- 12887700 TI - Oxidative phosphorylation by in situ synaptosomal mitochondria from whole brain of young and old rats. AB - Synaptosomes, isolated from the whole brain of young (3 months) and old (24 months) rats were used to study the major bioenergetic systems of neuronal mitochondria in situ, within the synaptosome. Approximately 85% of the resting oxygen consumption of synaptosomes from both young and old rats was a result of proton leak (and possibly other ion cycling) across the mitochondrial inner membrane. There were no significant differences between synaptosomes from the young and old rats in the kinetic responses of the substrate oxidation system, the mitochondrial proton leak and the phosphorylation system to changes in the proton electrochemical gradient. Flux control coefficients of 0.71, 0.27 and 0.02 were calculated for substrate oxidation system, phosphorylation system and the proton leak, respectively, at maximal ATP producing capacity in synaptosomes from young animals. The corresponding values calculated for synaptosomes from old animals were 0.53, 0.43 and 0.05. Thus substrate oxidation had greatest control over oxygen consumption at maximal phosphorylating capacity for synaptosomes from whole brain, with proton leak, having little control under maximal ATP producing capacity. The uncoupled rate of oxygen consumption, in the presence of the mitochondrial uncoupler, carbonyl cyanide p-trifluoromethoxyphenylhydrazone (FCCP), was significantly lower (p = 0.0124) in synaptosomes from old rats (6.08 +/- 0.42, n = 11) when compared with those from the young rats (7.87 +/- 0.48, n = 8). Thus, there is an impaired flux through the substrate oxidation system is synaptosomes from old rats, as compared to synaptosomes from the young animals. These in situ results may have important implications for the interpretation of theories that age-dependent impairment of mitochondrial energy production may result in increased susceptibility to neurodegeneration. PMID- 12887699 TI - Central catecholamine depletion inhibits peripheral lymphocyte responsiveness in spleen and blood. AB - Experimental and clinical evidence has demonstrated extensive communication between the CNS and the immune system. To analyse the role of central catecholamines in modulating peripheral immune functions, we injected the neurotoxin 6-hydroxydopamine (6-OHDA) i.c.v. in rats. This treatment significantly reduced brain catecholamine content 2, 4 and 7 days after injection, and in the periphery splenic catecholamine levels were reduced 4 days after treatment. Central catecholamine depletion induced an inhibition of splenic and blood lymphocyte proliferation and splenic cytokine production and expression (interleukin-2 and interferon-gamma) 7 days after injection. In addition, central treatment with 6-OHDA reduced the percentage of spleen and peripheral blood natural killer (CD161 +) cells, and T-cytotoxic (CD8 +) cells in peripheral blood. The reduction in splenocyte proliferation was not associated with a glucocorticoid alteration but was completely abolished by prior peripheral sympathectomy. These data demonstrate a crucial role of central and peripheral catecholamines in modulating immune function. PMID- 12887701 TI - Collapsin response mediator protein-2 accelerates axon regeneration of nerve injured motor neurons of rat. AB - The rat collapsin response mediator protein-2 (CRMP-2) is a member of CRMP family (CRMP-1-5). The functional consequence of CRMP-2 during embryonic development, particularly in neurite elongation, is relatively understood; however, the role in nerve regeneration is unclear. Here we examined the role of CRMP-2 during nerve regeneration using rat hypoglossal nerve injury model. Among the members, CRMP-1, CRMP-2, CRMP-5 mRNA expressions increased after nerve injury, whereas CRMP-3 and CRMP-4 mRNA did not show any significant change. In the N1E-115 cells, CRMP-2 has the most potent neurite elongation activity among the CRMP family members. In dorsal root ganglion (DRG) organ culture, CRMP-2 overexpression by adenoviral vector demonstrated substantial neurite elongation. On the other hand, CRMP-2 (DeltaC381), which acts as a dominant negative form of CRMP-2, inhibited neurite formation. Collectively, it would be plausible that CRMP-2 has potent nerve regeneration activity after nerve injury. We therefore examined whether CRMP-2 overexpression in the injured hypoglossal motor neurons accelerates nerve regeneration. A retrograde-tracer, Fluoro-Gold (FG), was used to evaluate the number of reprojecting motor neurons after nerve injury. CRMP-2-overexpressing motor neurons demonstrated the accelerated reprojection. The present study suggests that CRMP-2 has potent neurite elongation activity in nerve regeneration in vivo. PMID- 12887702 TI - Evidence for functional adenosine A3 receptors in microglia cells. AB - Adenosine exerts its effects through four subtypes of G-protein-coupled receptors (GPCRs): adenosine A1 and A3 receptors (A3R), which generally couple to Gi proteins and adenosine A2A and A2B receptors that activate Gs proteins. Though there is evidence for the expression of mRNA for the A3R in the central nervous system, evidence for functional receptors has depended on drugs with uncertain specificity. Here, we show that A3Rs mediating functional responses are present in microglia cells. By selectively stimulating the A3R in both primary mouse microglia cells and the N13 microglia cell line with the agonist Cl-IB-MECA, we have found a biphasic, partly Gi protein-dependent influence on the phosphorylation of the extracellular signal-regulated protein kinase 1/2 (ERK1/2). ERK1/2 activation was assessed by immunoblotting with phospho-specific antibodies. The involvement of the A3R in Cl-IB-MECA-induced ERK1/2 phosphorylation was confirmed by demonstrating that those effects are absent in primary mouse microglia cells isolated from mice lacking the gene for the A3R. PMID- 12887704 TI - All-digital image capture and whole-field analysis of ciliary beat frequency. AB - We hypothesized that a high-speed all-digital video imaging system, with computerized analysis, would precisely capture and measure ciliary beat frequency (CBF) and would shorten the time from data capture to data analysis. We compared a conventional analog video system with a new high-speed digital system we developed for CBF analysis. Using ciliated primary bovine bronchial epithelial cells we made simultaneous analog and digital CBF measurements of the same region of interest (ROI) while temperature was varied. This yielded nearly identical data over a wide range of frequencies (7-15 Hz) using either system. Unlike the digital system however, the analog system did not accurately detect CBF above 15 Hz (temperatures higher than 30 degrees C). We also compared ROI analysis with a new analysis algorithm we have named whole-field analysis (WFA). WFA measurement of CBF agreed with ROI and reduced operator time required to analyse data by more than 90% compared with the analog system. We conclude that all-digital computerized CBF analysis correlates closely with standard video methods, markedly speeds up data analysis and provides new ways, including WFA, to analyse entire fields of motile cilia simultaneously. We have termed this system 'Sisson Ammons Video Analysis' (SAVA). PMID- 12887705 TI - Conventional, confocal and two-photon fluorescence microscopy investigations of polymer-supported oxygen sensors. AB - Luminescence-based, polymer-supported oxygen sensors, particularly those based on platinum group complexes, continue to be of analytical importance. Commercial applications range from the macroscopic (e.g. aerodynamic investigations in wind tunnels, monitoring of oxygen concentration during fermentation, and measurement of biological oxygen demand) to the microscopic (e.g. imaging of oxygen in blood, tissue, cells and other biological samples). Problems hindering the design of improved oxygen sensors include non-linear Stern-Volmer calibration plots and the multi-exponentiality of measured lifetime decays, both of which are attributed primarily to heterogeneity of the sensor molecule in the polymer support matrix. Conventional, confocal and two-photon fluorescence microscopy have proven to be invaluable tools with which the microscale heterogeneity and response of luminescence-based oxygen sensors can be investigated and compared to the macroscopic response. Results obtained for three ruthenium(II) alpha-diimine complexes in polydimethylsiloxane polymer supports indicate the presence of unquenched microcrystals within the polymer matrix that probably degrade oxygen quenching sensitivity and linearity of the Stern-Volmer quenching plot. Two photon fluorescence microscopy proved most useful for imaging microcrystals within sensor films, and conventional microscopy allowed direct comparison between microscopic and macroscopic sensor response. The implications of the results in the rational design and mass production of luminescence-based oxygen sensors are significant. PMID- 12887706 TI - In vivo response to HA-polyhydroxybutyrate/polyhydroxyvalerate composite. AB - This study examined the morphological and compositional structure of bone-implant interfaces after in vivo implantation into the tibias of rabbits. The implants were composed of biodegradable polyhydroxybutyrate/polyhydroxyvalerate copolymer reinforced with synthetic hydroxyapatite (HA) particles. Optical and scanning electron microscopy techniques were used, including energy-dispersive X-ray analysis. The interface was found to be morphologically, biologically and chemically active throughout the period of study. There was a strong tendency to rebuild the bone structure at the interface after implantation, independent of the composition of the implant, but direct bone bonding with the implant depended on the bioactive nature of the interface, as represented by the HA particles. At all implantation times, lamellar bone formed at the interface and replaced degrading polymer matrix, while engulfing HA filler particles. In regions about 50-100 from the interface, the bone region displayed an osteon organization. Osteoblasts and osteocytes were identified throughout the interface region. The thickness of the newly formed bone significantly increased over the period of the experiment from about 130 microm at 1 month to about 770 microm at 6 months. Materials that behave in this manner may be useful in some bone replacement therapies. PMID- 12887707 TI - A simple approach to the determination of the crystallographic orientation: applications and accuracy. AB - A simple analytical solution for the crystallographic orientation is described. This method is based on one indexed Kikuchi pair in a known zone rather than the corresponding diffraction spots. The accuracy of this method is shown to be better than 0.1 degrees even for cases in which a zone axis deviates by a large angle (e.g. 10 degrees ) from the centre of the beam direction. This approach simplifies experiments because only one pair of Kikuchi lines and a zone axis are needed, and is especially suited when it is difficult or cumbersome to resolve a second pair of Kikuchi lines with sufficient accuracy. PMID- 12887708 TI - Statistical analysis of intramembranous particles using freeze fracture specimens. AB - We studied the point processes of intramembranous particles of mitochondrial membranes from HeLa cells using the freeze fracture technique. Three groups - under normal conditions, after exposition with rotenone, and after exposition with sodium acid - were compared. First, we used several summary statistics in order to study the two-dimensional point patterns of intramembranous particles within each group. Then, we compared the patterns in different groups by bootstrap tests using the K-function and the nearest neighbour distance function G(r). Estimation of the G-function provided significant results but no significant differences between groups were found using the classical K-function; estimation of G(r) should therefore not be omitted when studying observed planar point patterns. PMID- 12887709 TI - Wavelet-based image restoration for compact X-ray microscopy. AB - Compact water-window X-ray microscopy with short exposure times will always be limited on photons owing to sources of limited power in combination with low efficency X-ray optics. Thus, it is important to investigate methods for improving the signal-to-noise ratio in the images. We show that a wavelet-based denoising procedure significantly improves the quality and contrast in compact X ray microscopy images. A non-decimated, discrete wavelet transform (DWT) is applied to original, noisy images. After applying a thresholding procedure to the finest scales of the DWT, by setting to zero all wavelet coefficients of magnitude below a prescribed value, the inverse DWT to the thresholded DWT produces denoised images. It is concluded that the denoising procedure has potential to reduce the exposure time by a factor of 2 without loss of relevant image information. PMID- 12887710 TI - Preparation of site-specific cross-sections of heterogeneous catalysts prepared by focused ion beam milling. AB - Focused ion beam (FIB) milling offers a novel approach to preparation of site specific cross-sections of heterogeneous catalysts for examination in the transmission electron microscope (TEM). Electron-transparent sections can be obtained without the need to embed or grind the original sample. Because the specimen can be imaged in the FIB with submicrometre resolution before, during and after milling it is possible to select precisely the region from which the section is removed and to control the thickness of the section to within tens of nanometres. The ability to produce sections in this way opens the possibility of studying a range of catalyst systems that have previously been impossible to examine with the TEM. PMID- 12887711 TI - Four-dimensional imaging and computer-assisted track analysis of nuclear migration in root hairs of Arabidopsis thaliana. AB - Nuclear migration is a fundamental mechanism necessary for the proper growth and development of many eukaryotic organisms. In this study root hairs of Arabidopsis thaliana were used as a research model to gain insight into the dynamics of nuclear migration. Root hairs are long tubular outgrowths of epidermal cells and are responsible for the uptake of water and nutrients. During the development of root hairs, the nucleus migrates into the hair after the bulge is formed. The position of the nucleus relative to the tip plays an essential role in the growth process. However, what is happening to the nucleus in full-grown root hairs is still unclear. To study nuclear dynamics in living root hair cells, stably transformed plants with the fusion proteins Histone2B-YFP and NLS-GFP-GUS were used. Four-dimensional confocal laser scanning microscopy made it possible to monitor the exact position of the nucleus in different root hairs. To analyse the sequential positions of the nuclei in the root hairs, a new computer-assisted method was developed. After track analysis a number of parameters could be extracted from the movies, such as the average speed, the amplitude, direction factor and the range of movement in the root hairs. Our results show that nuclei do not reach a final position in full-grown root hairs and this sustained movement seems to be more similar in root hairs lying close to each other. Moreover, with this methodology it could be quantitatively demonstrated that the integrity of actin is necessary for nuclear movement. PMID- 12887712 TI - Correction of autofocusing errors due to specimen tilt for automated electron tomography. AB - Transmission electron microscopy images acquired under tilted-beam conditions experience an image shift as a function of defocus settings - a fact that is exploited as a method for defocus determination in most of the automated tomography data collection systems. Although the method was shown to be highly accurate for a large variety of specimens, we point out that in its original design it can strictly only be applied to images of untilted samples. The application to tilted samples and thus in automated electron tomography is impaired mainly due to a defocus change across the images, resulting in reduced accuracy. In this communication we present a method that can be used to improve the accuracy of the basic autofocusing procedures currently used in systems for automated electron tomography. PMID- 12887718 TI - Brief history of low-density lipoprotein apheresis. AB - Low-density lipoprotein (LDL) apheresis is a technology used to remove LDL from hypercholesterolemic patients. The technique includes both specific and non specific removal of LDL such as exchange of the whole plasma and selective adorption of LDL. The therapy is mainly used for homozygotes and in severe cases of heterozygotes of familial hypercholesterolemia (FH) (LDL receptor deficiency). Since HMG-CoA reductase inhibitors, statins, were introduced in late 1980s, the number of patients who required LDL apheresis decreased, in particular for the heterozygous FH patients with more recent powerful statins. Modern LDL apheresis technology is very sophisticated and perhaps expensive, while the number of the patients is very limited. Several companies developed the instruments for this procedure, but found the market is limited. Nevertheless, these companies are expected to remain in the market for an ethical reasons, as the patients using their LDL apheresis machines are entirely dependent on them for their life. PMID- 12887719 TI - Low-density lipoprotein apheresis: an overview. AB - Atherosclerosis with myocardial infarction, stroke, and peripheral cellular disease still maintains its position at the top of morbidity and mortality statistics in industrialized nations. Established risk factors widely accepted are smoking, arterial hypertension, diabetes mellitus, and central obesity. Furthermore, there is a strong correlation between hyperlipidemia and atherosclerosis. The prognosis of patients suffering from severe hyperlipidemia, sometimes combined with elevated lipoprotein (a) (Lpa) levels, and coronary heart disease (CHD) refractory to diet and lipid-lowering drugs is poor. For such patients, regular treatment with low-density lipoprotein (LDL) apheresis is the therapeutic option. Today, there are four different LDL apheresis systems available: immunoadsorption, heparin-induced extracorporeal LDL/fibrinogen precipitation, dextran sulfate LDL adsorption and LDL hemoperfusion. Regarding the different LDL apheresis systems used, there is no significant difference with respect to the clinical outcome or concerning total cholesterol, LDL, high density lipoprotein (HDL), or triglyceride concentrations. With respect to elevated Lpa levels, however, the immunoadsorption method seems to be the most effective. In 45 patients (25 women, 20 men) suffering from familial hypercholesterolemia resistant to diet and lipid lowering drugs, low-density lipoprotein (LDL) apheresis was performed over 95.6 +/- 44.7 months. Four different systems (Liposorber, 32 of 45, Kaneka, Osaka, Japan; Therasorb, 6 of 45, Baxter, Munich, Germany; Lipopak, 2 of 45, Pocard, Moscow, Russia; and Dali, 5 of 45, Fresenius, St. Wendel, Germany) were used. With all methods, average reductions of 57% for total cholesterol, 55.9% for LDL, 75.8% for lipoprotein a (Lpa), and 45.9% for triglycerides, and an average increase of 14.3% for HDL were reached. Severe side-effects such as shock or allergic reactions were very rare (0.3%) in all methods. In the course of treatment, an improvement in general well being and increased performance were experienced by 44 of 45 patients. The present data demonstrate that treatment with LDL apheresis of patients suffering from familial hypercholesterolemia resistant to maximum conservative therapy is very effective and safe even in long-term application. PMID- 12887720 TI - The HELP system for the treatment of atherothrombotic disorders: a review. AB - Given the importance of atherothrombotic disorders for the public health system, and the known limitations of conventional treatment on one hand and the compelling biochemical evidence and long-term safety of HELP (Heparin-mediated Extracorporeal LDL/Fibrinogen Precipitation) apheresis on the other hand, this approach provides a most valuable tool for further medical research and treatment of the various atherothrombotic and microcirculatory disorders. The present contribution reviews the recent developments in chronic and single application of apheresis in cardiology with particular emphasis on the newly discovered therapeutic possibilities for myocardial infarction, stroke, and after coronary artery bypass grafting. PMID- 12887721 TI - Long-term effect of low-density lipoprotein apheresis in patients with homozygous familial hypercholesterolemia. AB - Patients that are homozygous for familial hypercholesterolemia (FH) exhibit severe hypercholesterolemia, cutaneous and tendon xanthomas and premature atherosclerosis beginning in childhood. They are resistant to drug therapy and low-density lipoprotein (LDL) apheresis is the practical treatment. Here we review the technique of LDL apheresis treatment, the long-term effects of LDL apheresis, the effect of apheresis on pregnancy, and the drugs that have proven beneficial in patients with homozygous FH. We also record our experiences of treating eight homozygous FH patients using the LDL apheresis treatment. Among the eight patients, one has been free from cardiovascular disease and two patients have each regressed once. In two patients, aortic valve stenosis developed and the other two patients died for acute myocardial infarction. Furthermore, two patients delivered healthy babies in spite of coronary artery disease. Thus, LDL apheresis therapy has the possibility of preventing the progression of atherosclerosis, but the prognosis assessed by long-term observation is still not satisfactory. A recent clinical trial showed some efficacy of the combination therapy of LDL apheresis and atorvastatin for reducing serum cholesterol levels in homozygous FH, suggesting that this combination therapy may be useful for prevention of atherosclerosis in patients homozygous for FH. PMID- 12887722 TI - Long-term effect of low-density lipoprotein apheresis in patients with heterozygous familial hypercholesterolemia. AB - Clinical efficacy and safety of the therapeutic tool which directly removes LDL particles from circulation (LDL apheresis) have already been established in the treatment for refractory hypercholesterolemia in patients with familial hypercholesterolemia (FH). Two clinical studies with event-based assessment have demonstrated remarkably beneficial outcomes of long-term LDL apheresis using dextran sulfate cellulose columns plus adjunctive cholesterol-lowering drug therapy in the prevention of cardiovascular events in heterozygous FH with coronary artery disease. The results of several studies with angiographic and ultrasound-based assessment indicate a possible role for LDL apheresis in restructuring and stabilization of atherosclerotic lesions. These clinical improvements caused by LDL apheresis in heterozygous FH support the efficacy and importance of aggressive cholesterol-lowering therapy for secondary prevention of atherosclerotic cardiovascular disease in hypercholesterolemic patients. PMID- 12887723 TI - Low-density lipoprotein apheresis in the prevention of recurrent coronary heart disease: a review. AB - This article summarizes the clinical evidence for low-density lipoprotein (LDL) apheresis to improve prognosis of patients with coronary heart disease. Regarding clinical events or angiographic parameters, most trials have revealed favorable effects on coronary artery disease. In the studies with a clinical endpoint, approximately 50% reduction in incidence of major coronary events was achieved by LDL apheresis. Similarly, in the studies with a angiographic parameter, coronary atherosclerosis was suppressed or regressed by LDL apheresis. The mechanisms of these effects by LDL apheresis are attributed to improvement in coronary flow reserve, which is dependent upon increases in hemo-rheological factor, nitric oxide, vasoactive substances or decrease of oxidized LDL. These new insights into mechanisms will expand the indication of LDL apheresis to other pathophysiological conditions of coronary artery disease and other disorders. PMID- 12887724 TI - Low-density lipoprotein apheresis in the treatment of peripheral arterial disease. AB - Low-density lipoprotein (LDL) apheresis is performed in patients with homozygous familial hyper-cholesterolemia who lack LDL receptors and with heterozygous familial hypercholesterolemia who are LDL receptor deficient with documented, symptomatic coronary artery disease who are resistant to diet changes and maximum drug therapy. LDL apheresis can reduce or abolish anginal symptoms and improve coronary lesions. Several reports reveal the improvement of insufficient peripheral blood flow. By extensively removing blood LDL and changing coagulation factors and various vasoactive substances, LDL apheresis improves blood rheology and thereby peripheral circulation. It seems worth trying on all patients with arteriosclerotic lesions, even if they are normocholesterolemic. PMID- 12887725 TI - Changes in oxidative stress and microcirculation by low-density lipoprotein apheresis. AB - Low-density lipoprotein apheresis (LDLA) leads to an improvement of microcirculation during the very early stages of treatment, and continued treatment may produce antiatherogenic effects in patients with peripheral arterial disease (PAD). Suppression of oxidative stress, improvement of endothelial functions and alteration in the action of vasoactive compounds may occur with the improvement of the rheological property of blood as a result of aggressive removal of atherogenic factors including LDL, possibly resulting in the suppression of development of atherosclerosis. As these effects of LDLA may ameliorate not only PAD but also ischemia in other organs, it is suggested that repeated LDLA prevents the progression of atherosclerotic diseases and probably improves the long-term prognosis of patients with PAD. PMID- 12887726 TI - The effect of apheresis on adhesion molecules. AB - The adhesion molecules on the leukocytes and the endothelial cells mediate interaction between their cells. The plasma levels of soluble adhesion molecules increase in patients with ischemic heart disease, atherosclerotic aortic disease. Hypercholesterolemia is one of risk factors for atherosclerosis, and it is considered that the expression of adhesion molecules in endothelial cells is related to the development of atherosclerosis. Low-density lipoprotein (LDL) apheresis has been applied to patients with hypercholesterolemia. LDL apheresis may have an effect on adhesion molecules in patients with hypercholesterolemia. PMID- 12887727 TI - Changes in C-reactive protein plasma levels during low-density lipoprotein apheresis. AB - C-reactive protein (CRP) is one of the important risk factors for atherosclerosis, and its serum level is lowered by popular cholesterol-lowering drugs, statins. This study was undertaken to examine the changes of CRP levels during dextran-sulfate (DS) low-density lipoprotein (LDL) apheresis. In 15 apheresis sessions in seven patients with severe hypercholesterolemia (four men and three women, aged between 36 and 70 years), changes in CRP levels were examined. The efficiency in adsorption of CRP with DS column was evaluated by measuring CRP levels in both pre- and post-column plasma. In one patient, the effect of repeated apheresis sessions on CRP preapheresis levels was examined. The changes in interleukin (IL)-6 plasma levels were also examined in six sessions. Although IL-6 levels after 3,000 mL-plasma treatment rose to 170% of preapheresis levels, CRP levels decreased significantly (from 1.91 +/- 0.49 mg/L to 0.89 +/- 0.24, P < 0.01). C-reactive protein was almost completely adsorbed by the DS column and CRP preapheresis levels were decreased gradually by repetition of apheresis. CRP, a novel risk factor of atherosclerosis, was effectively removed by DS-LDL apheresis. The decrease in CRP plasma levels may be involved in prevention of atherosclerotic vascular diseases due to DS-LDL apheresis. PMID- 12887728 TI - Apheresis in the treatment of cholesterol embolic disease. AB - Cholesterol embolic disease is a devastating complication of atherosclerosis. Universally recommended treatment is lacking thus far. Recent data suggest that a therapeutic protocol aimed at specifically combating three causes of mortality, recurrent bouts of cholesterol embolism, cardiac failure, and cahexia, were associated with a favorable clinical outcome. As for drug therapy, corticosteroid has been reported to be beneficial in reducing local and general inflammatory responses. Concerning apheresis, combined therapy consisting of plasma exchange and low to intermediate-dose corticosteroid therapy has been shown to be effective in multivisceral cholesterol embolism. Low density lipoprotein (LDL) apheresis has been reported to be beneficial for cholesterol embolism-induced damage to the skin and brain. PMID- 12887729 TI - Hypercholesterolemic valvulopathy: an aspect of malignant atherosclerosis. AB - Hypercholesterolemia affects not only the coronary artery, but also the aortic root, particularly the aortic valve. Aortic stenosis is critical in the prognosis for most patients with homozygous familial hypercholesterolemia (FH) and some heterozygous FH patients who result in aortic valve replacement (AVR). Histopathological examination of their valves shows lipid deposition, inflammatory cell infiltration and calcification in the aortic cusps. These pathological findings are common in non-FH patients with AVR. In homozygous FH patients, the aortic valves are injured by extreme hypercholesterolemia in a relatively short period of time, whereas in heterozygous FH patients with additional risk factors, the damage to the valves occurs over an intermediate time period. In the non-FH population with several risk factors, an underlying raised serum cholesterol level results in gradual impairment over a number of years. Aortic valvular dysfunction caused by hypercholesterolemia, which we termed hypercholesterolemic valvulopathy is recognized to be a life-threatening complication as the primary clinical characteristic of malignant atherosclerosis. PMID- 12887730 TI - Rheopheresis in patients with ischemic diabetic foot syndrome: results of an open label prospective pilot trial. AB - Rheopheresis is a specific application of membrane differential filtration, synonymous with double filtration plasmapheresis, for extracorporeal hemorheotherapy. Safety and efficacy of Rheopheresis for wound healing and skin oxygenation were investigated in patients with ischemic diabetic foot syndrome. Eight patients with type 2 diabetes mellitus and non-healing foot ulcers caused by severe ischemic diabetic foot syndrome were treated by a series of seven Rheopheresis sessions in a time span of 11 weeks. Wound healing had not been detectable under conditions of standardized wound care during at least 2 months. Wound status was classified by its morphology, severity and location, according to the criteria of Wagner. Transcutaneous oxygen pressure (tcPO2), laser Doppler flowmetry and vital capillary microscopy were repeatedly performed to monitor the effects of the Rheopheresis treatment series on microcirculation and skin blood flow. Laboratory parameters of blood rheology, endothelial function and inflammatory state were measured in addition to safety parameters. In four patients (baseline Wagner stage 2), Rheopheresis accelerated wound healing of foot ulcers and was associated with an improvement of Wagner stage and a pronounced increase in tcPO2. In two patients (baseline Wagner stage 2), wound healing was unchanged but mean tcPO2 increased, allowing successful minor amputation. Values of tcPO2 remained stable and enhanced for the 3 months follow up period. In two patients (baseline Wagner stage 4 or 5), no improvements in foot lesions were observed within the treatment period. As an adjunct therapeutic option, Rheopheresis may preserve a functional lower extremity, delay amputation or reduce the extent of amputation. PMID- 12887731 TI - The sequential organ failure assessment score as a useful predictor for estimating the prognosis of systemic inflammatory response syndrome patients being treated with extracorporeal blood purification. AB - Systemic inflammatory response syndrome (SIRS) is a major cause of morbidity and mortality in critically ill patients. Extracorporeal blood purification procedures are becoming important for treating these patients. However, the cost of these procedures is high. Therefore, a prognostic marker would be helpful. To establish the reliability of the Sequential Organ Failure Assessment (SOFA) score as a prognostic indicator, we evaluated daily changes in the SOFA score of 40 SIRS patients who needed blood purification procedures such as continuous renal replacement therapy (CRRT), endotoxin adsorption, bilirubin adsorption, and/or plasma exchange. Twenty patients survived and 20 died. Although the baseline scores of the two groups (survivors and non-survivors) did not differ, both the maximum value of the SOFA score and the DeltaSOFA score (the difference between the maximum SOFA and baseline SOFA scores) were significantly higher in the non survivor group. The mortality rate among patients with a maximum SOFA score greater than or equal to 18 or a DeltaSOFA score greater than or equal to 3 was higher than for the rest of the patients. The changes in the SOFA score correlated well with the outcomes of the SIRS patients. The maximum SOFA score and the DeltaSOFA score are therefore likely to be useful prognostic markers. PMID- 12887732 TI - Oxygen uptake efficiency slope as monitoring tool for physical training in chronic hemodialysis patients. AB - The purpose of this study was to clarify the utility of oxygen uptake efficiency slope (OUES) as a monitoring tool, and to investigate the effects of physical training in chronic hemodialysis (HD) patients. Seventeen patients (Trained Group) received physical training 2-3 times per week for 20 weeks at the intervals between exercise tests. Patients underwent a combination training of bicycle ergometry, walking and jogging for 30 min duration. The intensity of physical training was adjusted to maintain the exercising heart rate at between 50 and 60% of the peak heart rate. Twelve patients (Control Group) lived without physical training throughout the 20 weeks. Both the Groups received the symptom limited exercise tests before and after the 20 week physical training. Minute ventilation (VE), carbon dioxide output (VCO2) and oxygen uptake (VO2) were continuously measured during the exercise tests. Oxygen uptake efficiency slope was derived from the logarithmic relation between VO2 and VE during an incremental exercise test. In the Trained Group, OUES after physical training (30.1 +/- 5.8) was significantly (P < 0.01) higher than that before physical training (25.2 +/- 2.6), while in the Control Group, OUES did not change in this study period of 20 weeks. In the Trained Group, changes in OUES correlated with those in the maximum oxygen uptake (r = 0.78, P < 0.001) and the anaerobic threshold (r = 0.61, P < 0.01). It was suggested that OUES was applicable as a monitoring tool for cardiorespiratory functional reserve during physical training in HD patients. PMID- 12887733 TI - A case of advanced primary biliary cirrhosis treated with granulocyte and monocyte apheresis. AB - Generally, the most effective treatment for advanced primary biliary cirrhosis (PBC) is liver transplantation, but adjunct therapies are needed. We report here a first case of advanced PBC treated with a new immunotherapy, granulocyte and monocyte apheresis (GCAP). A column (Adacolumn, Japan Immunoresearch Laboratory Takasaki, Japan) was filled with cellulose acetate beads to selectively adsorb granulocytes and monocytes/macrophages. A 49-year-old woman was diagnosed with PBC in 1987. In June 2001, steroid pulse therapy and adjuvant fresh frozen plasma was given for moderate jaundice but without success. In July, as total bilirubin rapidly increased, treatment with GCAP was started and succeeded in suppressing the rapid deterioration of total bilirubin (value changes after each of four applications: 15.4-->14.0, 27.2-->25.1, 25.8-->24.0, 25.7-->23.7 mg/dL) and improving prothrombin time (16.4-->14.5 s). Although GCAP therapy did not prevent a fatal outcome, it suppressed rapid deterioration of jaundice and increased quality of life for a month. PMID- 12887734 TI - Reduced-folate carrier (RFC) is expressed in placenta and yolk sac, as well as in cells of the developing forebrain, hindbrain, neural tube, craniofacial region, eye, limb buds and heart. AB - BACKGROUND: Folate is essential for cellular proliferation and tissue regeneration. As mammalian cells cannot synthesize folates de novo, tightly regulated cellular uptake processes have evolved to sustain sufficient levels of intracellular tetrahydrofolate cofactors to support biosynthesis of purines, pyrimidines, and some amino acids (serine, methionine). Though reduced-folate carrier (RFC) is one of the major proteins mediating folate transport, knowledge of the developmental expression of RFC is lacking. We utilized in situ hybridization and immunolocalization to determine the developmental distribution of RFC message and protein, respectively. RESULTS: In the mouse, RFC transcripts and protein are expressed in the E10.0 placenta and yolk sac. In the E9.0 to E11.5 mouse embryo RFC is widely detectable, with intense signal localized to cell populations in the neural tube, craniofacial region, limb buds and heart. During early development, RFC is expressed throughout the eye, but by E12.5, RFC protein becomes localized to the retinal pigment epithelium (RPE). CONCLUSIONS: Clinical studies show a statistical decrease in the number of neural tube defects, craniofacial abnormalities, cardiovascular defects and limb abnormalities detected in offspring of female patients given supplementary folate during pregnancy. The mechanism, however, by which folate supplementation ameliorates the occurrence of developmental defects is unclear. The present work demonstrates that RFC is present in placenta and yolk sac and provides the first evidence that it is expressed in the neural tube, craniofacial region, limb buds and heart during organogenesis. These findings suggest that rapidly dividing cells in the developing neural tube, craniofacial region, limb buds and heart may be particularly susceptible to folate deficiency. PMID- 12887735 TI - Human cloning laws, human dignity and the poverty of the policy making dialogue. AB - BACKGROUND: The regulation of human cloning continues to be a significant national and international policy issue. Despite years of intense academic and public debate, there is little clarity as to the philosophical foundations for many of the emerging policy choices. The notion of "human dignity" is commonly used to justify cloning laws. The basis for this justification is that reproductive human cloning necessarily infringes notions of human dignity. DISCUSSION: The author critiques one of the most commonly used ethical justifications for cloning laws - the idea that reproductive cloning necessarily infringes notions of human dignity. He points out that there is, in fact, little consensus on point and that the counter arguments are rarely reflected in formal policy. Rarely do domestic or international instruments provide an operational definition of human dignity and there is rarely an explanation of how, exactly, dignity is infringed in the context reproductive cloning. SUMMARY: It is the author's position that the lack of thoughtful analysis of the role of human dignity hurts the broader public debate about reproductive cloning, trivializes the value of human dignity as a normative principle and makes it nearly impossible to critique the actual justifications behind many of the proposed policies. PMID- 12887737 TI - [The neuroprotection of resveratrol in the experimental cerebral ischemia]. AB - OBJECTIVE: To study the effect of resveratrol on cerebral infarct volume, cerebral edema as well as the expression of nuclear fac for (NF-kappaB) p65 in the experimental permanent middle cerebral artery occlusion. METHODS: A middle cerebral artery occlusion model was constructed in 20 Wistar rats with Longa's method. Then the 20 rats were divided into 4 groups: control group, and 3 resveratrol groups (resveratrol of the concentrations of 10(-8) g/kg, 10(-7) g/kg, and 10(-6) g/kg were injected intravenously) (group 1- 4). The area of infarct was examined by 2,3,5-triphenyltetrazolium chloride (TTC) staining. The water content was calculated by dry and wet weight method. The expression of NF kappaB p65 was investigated with immunohistochemistry. RESULTS: The ischemic volume was 241 +/- 36, 222 +/- 39, 126 +/- 16, and 128 +/- 19 * in the groups 1 - 4 respectively, and the volume ratio was 0.49 +/- 0.04, 0.47 +/- 0.04, 27 +/- 0.02, and 0.43 +/- 0.02 respectively. The brain water content was 83.5% +/- 1.5%, 82.9% +/- 1.4%, 79.5% +/- 1.5%, and 79.9% +/- 1.4% in groups 1 - 4 with a significant difference between 10(-7) g/kg resveratrol group and pure ischemia group and 10(-6) g/kg resveratrol group and pure ischemia group (both P < 0.05). NF-kappaB p65 was not expressed in the normal control group and sham operation group, and was expressed 2 hours after pure ischemia. The expression of NF-kappaB p65 positive cells was the most significant 12 hours after the pure ischemia and began to decrease 24 hours after ischemia. CONCLUSION: Resveratrol has a neuroprotective effect on the ischemic brain. This effect is related to the inhabitation of expression of NF-kappaB p65. PMID- 12887738 TI - [The relationship of apolipoprotein H gene polymorphism with stroke]. AB - OBJECTIVE: To study the association between apolipoprotein H (apoH) gene polymorphism and cerebral apoplexy among the Hans in Changsha, Hunan Province. METHODS: Polymorphisms in exon 3 of apoH gene was determined by PCR-single strand conformation polymorphism (PCR-SSCP) analysis and DNA sequencing among 260 patients with stroke of Han nationality in Changsha, including 130 cerebral hemorrhage (CH) patients, of which 50 were with a family history of CH, and 79 with hypertension and 51 without hypertension, and 130 cerebral infarction (CI) patients, of which 50 were with a family history of CI, and 66 with hypertension and 64 without hypertension; 20 members of stroke pedigrees, and 100 healthy controls. RESULTS: G341A (Ser88Asn) polymorphism in exon 3 of apoH gene was found in Changsha Hans. The frequency of A allele was 0.127 in the CH group, especially in CH patients with a family history of stroke or hypertension, significantly higher than that in control group (0.055, P < 0.05). The frequency of A allele was also significantly higher in members of CH pedigrees than in controls. No association between G341A (Ser88Asn) polymorphism and CI was found. CONCLUSION: A allele in G341A (Ser88Asn) polymorphism may be a genetic risk factor of CH among Changsha Hans, especially the CH patients with hypertension and those with a family history of hypertension. PMID- 12887736 TI - Interferon-alpha/beta receptor-mediated selective induction of a gene cluster by CpG oligodeoxynucleotide 2006. AB - BACKGROUND: Oligodeoxynucleotides containing unmethylated CpG motifs (CpG ODN) are known to exert a strong adjuvant effect on Th1 immune responses. Although several genes have been reported, no comprehensive study of the gene expression profiles in human cells after stimulation with CpG ODN has been reported. RESULTS: This study was designed to identify a CpG-inducible gene cluster that potentially predicts for the molecular mechanisms of clinical efficacy of CpG ODN, by determining mRNA expression in human PBMC after stimulation with CpG ODN. PBMCs were obtained from the peripheral blood of healthy volunteers and cultured in the presence or absence of CpG ODN 2006 for up to 24 hours. The mRNA expression profile was evaluated using a high-density oligonucleotide probe array, GeneChip. Using hierarchical clustering-analysis, out of a total of 10,000 genes we identified a cluster containing 77 genes as having been up-regulated by CpG ODN. This cluster was further divided into two sub-clusters by means of time kinetics. (1) Inflammatory cytokines such as IL-6 and GM-CSF were up-regulated predominantly 3 to 6 hours after stimulation with CpG ODN, presumably through activation of a transcription factor, NF-kappaB. (2) Interferon (IFN)-inducible anti-viral proteins, including IFIT1, OAS1 and Mx1, and Th1 chemoattractant IP 10, were up-regulated predominantly 6 to 24 hours after stimulation. Blocking with mAb against IFN-alpha/beta receptor strongly inhibited the induction of these IFN-inducible genes by CpG ODN. CONCLUSION: This study provides new information regarding the possible immunomodulatory effects of CpG ODN in vivo via an IFN-alpha/beta receptor-mediated paracrine pathway. PMID- 12887739 TI - [Effect of intraischemic mild hypothermia on interleukin-1beta and monocyte chemoattractant protein-1 contents in ischemic core of rat cortex after transient focal cerebral ischemia]. AB - OBJECTIVE: To investigate the effect of intraischemic mild hypothermia on the protein levels of interleukin (IL)-1beta and monocyte chemoattractant protein (MCP)-1 in the ischemic core of rat cortex after transient focal cerebralischemia. METHODS: Eighty male Wistar rats were randomly divided into normothermic (37 degrees C) and mild hypothermic (32 - 33 degrees C) groups. The normothermic group was redivided into six subgroups of 8 rats: sham operation, ischemia for 2 hours without reperfusion, and reperfusion for 6 hours, 16 hours, 24 hours, and 48 hours respectively after ischemia; and the mild hypothermic group was redivided into 4 group with 8 rats: reperfusion for 6, 16, 24, and 48 hours. The rats except those in the sham operation subgroup were subjected to right middle cerebral artery occlusion by insertion a specially prepared nylon filament for two hours. Ice bag was used to lower the brain temperature and anal temperature soon after ischemia to 32.0 - 33.0 degrees C within 10 minutes in the mild hypothermic subgroups. The brain and anal temperature remained at 37.0 - 37.5 degrees C in all normothermic subgroups. Then the rats were killed 0, 6, 16, 24 and 48 hours after reperfusion respectively and their brains were taken out to examine the size of brain infarct by 2,3,5-triphenyltetrazolium chloride (TTC) staining reaction. The protein levels of IL-1beta and MCP-1 in the cortical ischemic core were measured by ELISA. RESULTS: No significant change of IL-1beta protein level was found in the cortical ischemic cores at any time point after reperfusion among the normothermic subgroups. The IL-1beta protein levels at different time points were not significantly different between the intraischemic mild hypothermia subgroups and the normothermic subgroups (all P > 0.05). The MCP 1 protein level in the cortical ischemic cores of the normothermic subgroups began to increase since the 6th hour afer reperfusion (22.5 +/- 8.7 ng x g tissue(-1), 17 times that in the sham operation samples, P < 0.05), peaked in 48 hours (110.9 +/- 47.0 ng x g tissue(-1), 83.7 times that in the sham operation sample, P < 0.001). The protein level of MCP-1 in the mild hypothermic subgroups was 8.7 +/- 7.6 ng x g tissue(-1) 6 h after reperfusion (P < 0.005 in comparison with those in sham operation subgroup and ischemia subgroup) and was 56.0 +/- 40.3 ng x g tissue(-1), 48 hours after reperfusion (P < 0.05) incomparison with those in the normothermic subgroups). The sizes of cortical infarct at different time points in the mild hypothermic subgroups were significantly smaller than those in the normothermic subgroups (P < 0.05). CONCLUSION: Mild hypothermia reduces the level of MCP-1 in the cortex after cerebral ischemia/reperfusion which may be one of the important mechanisms of the neuroprotective effects of mild hypothermia. PMID- 12887740 TI - [Immunohistochemical study on central nervous system cavernous hemangiomas]. AB - OBJECTIVE: To explore the histological structure, angiogenesis, and proliferative activity of central nervous system cavernous hemangiomas. METHODS: 70 surgical samples of central nervous system cavernous hemangiomas and 20 normal brain vessel samples from patients of epilepsy and open craniocerebral trauma were stained immunohistochemically with CD34, a-SMA; VEGF, Flt-1; and TGFa, Ki67 respectively. A comparison analysis was made according to the expression intensity. RESULTS: CD34 and a-SMA were expressed in all the normal control brain vessel tissues in a manner of obvious and continuous staining. VEGF, Flt-1 and TGFa were not expressed obviously in the normal brain tissues. 47 and 50 out of the 70 cavernous hemangioma specimens were positively stained for CD34 and a-SMA respectively, and their expression was less continuous. 68 and 44 out of the 70 cavernous hemangioma specimens were positively stained for VEGF and Flt-1 respectively with diffuse distribution. 68 cavernous hemangioma specimens were positively stained for TGF-a. A significant difference in expression intensity was found for the above 5 factors between the normal control brain tissue and cavernous hemangiomas (all P < 0.05). No expression of Ki67 was detected in all samples. CONCLUSION: The biological characteristics of cavernous hemangiomas are mainly relevant to the immaturity of the vessel wall. A series of angiogenic factors play an important role in the development of the lesion. The proliferative activity of the cavernous hemangiomas needs to be studied further. PMID- 12887741 TI - [Identification of phenotype and Interferon-alpha-producing capability of circulating type II dendritic cells and its clinical implication in HBV-infected patients]. AB - OBJECTIVE: To investigate the number, phenotype, and interferon-alpha (INF-alpha) of type II dendritic cells (DC2) in persons with hepatitis B and evaluate the role of DC2 subset in the immunopathogenesis of chronic HBV infection. METHODS: Peripheral blood was extracted from 103 hepatitis B (HB) virus-infected persons, including 11 cases of HB virus (HBV)-infected persons, 11 cases of acute HB, 81 cases of chronic HB, and 11 cases of asymptomatic HBV infection, and 25 healthy blood donors used as controls. Flow cytometry was used to calculate the number and the phenotype of circulating DC2. Ultraviolet-inactivated herpes simplex virus (HSV)-1 was added into the suspension of peripheral blood mononuclear cells (PBMCs) and then co-cultured for 24 hours to stimulate the production of INF alpha by DC2 that was examined by ELISA assay. RESULTS: The number of DC2 in patients with chronic HB was 3.3 +/- 1.0 10(6)/L, significantly lower than that in the healthy controls (7.2 +/- 2.4 10(6)/L, P < 0.01). However, the number of DC2 was not significantly different between any other groups. The proportion of GS2 to PBMCs in the patients with chronic HB was 1.12 +/- 1.13 approximately 0.22 +/- 0.10, all significantly lower than that in the healthy controls (0.32% +/- 0.13%, P < 0.01). However, the proportion of GS2 to PBMCs was not significantly different between any other groups. The decrease of number of DC2 and that of proportion of DC2 to PBMCs in patients with chronic HB were related with the progress of disease. The INF-alpha concentration in the suspensions of PBMCs of different groups without stimulation by HSV-1 were low and there was no significant difference in INF-alpha concentration between different groups. The INF-alpha concentration in the suspension of PBMCs of healthy controls was 789 +/ 82 pg/ml, significantly higher than those of the patients with acute HB (161 +/- 36 pg/ml) and the patients with chronic HB (183 +/- 113 pg/ml, 147 +/- 39 pg/ml, and 156 +/- 39 pg/ml, all P < 0.05). However, there was no significant difference between the patient groups (all P > 0.05). CONCLUSION: The number and INF-alpha producing function of DC2, and the numbers of NK cells and CD8+ T cells in peripheral blood of patients with chronic HB decrease significantly, which results the deficiency of HBV-specific immune response. PMID- 12887742 TI - [Clinical manifestation and prognosis of myocardial bridge]. AB - OBJECTIVE: To study the clinical manifestation, angiographic features, and prognosis of myocardial bridge. METHODS: A retrospective analysis was made on the data of the clinical manifestation, coronary angiography, and prognosis of 35 patients with myocardial bridge, 29 males and 6 females, with an average age of 52.0 +/- 9.5 years, out of 2 871 patients undergoing coronary angiography 1 January 1996 - 20 February 2001. RESULTS: The detection rate of myocardial bridge, mostly in the middle or distal parts of left anterior descending branch and 24 being isolated myocardial bridge, was 1.22% in coronary angiography. There was a significant difference in the extent of diameter stenosis during systolic stage between the group with atherosclerosis (68% +/- 15%, n = 15) and the group without atherosclerosis (54% +/- 14%, n = 20) in the vessel proximal to myocardial bridge (P < 0.01). The systolic diameter stenosis was more severe in the abnormal ECG group (63% +/- 13%, n = 12) than in the normal ECG group (54% +/ 14%, n = 12), P < 0.05. However, the systolic stenosis extent of myocardial bridge in the patients with typical angina pectoris (58% +/- 15%, n = 11) was not significantly different from that in the patients with atypical angina pectoris (54% +/- 15%, n = 13). The systolic stenosis extent of myocardial bridge were 69% +/- 9% (n = 7) and 58% +/- 16% (n = 26) in the patients with and without left ventricular wall hypertrophy respectively (P = 0.09). No malignant event occurred during the follow-up period of 3 - 50 months. CONCLUSION: (1) The more severe the extent of systolic diameter stenosis, the more severe the myocardial ischemia and the more the possibility of abnormal ECG. (2) Myocardial bridge tends to promote or accelerate the atherosclerosis of the vessels proximal to it. (3) Left ventricular wall hypertrophy may promote the formation of myocardial bridge clinically. (4) The prognosis of myocardial bridge is good. PMID- 12887743 TI - [Expression of cyclin A in adult patients with acute leukemia]. AB - OBJECTIVE: To investigate the clinical significance of cyclin A expression in adult patients with acute leukemia (AL). METHODS: 4 ml bone marrow was extracted from 100 AL patients and 10 normal controls to isolate the mononuclear cells (MNCs). The cyclin A mRNA levels in these MNCs were measured by RT-PCR. The cyclin A protein level and cell cycle in 75 randomly selected AL patients and 10 normal controls were examined by flow cytometric analysis. RESULTS: (1) The distribution of cyclin A protein was normal in cell cycle in AL patients. The positive rates of cyclin A protein and mRNA were 66.7% and 59%, significantly higher than those in the normal controls (0 and 1.4% respectively, both P < 0.01). The median expression levels of cyclin A protein and mRNA of cyclin A in AL patients were 18.5% and 0.539 +/- 0.490 respectively, significantly higher than those in the normal controls (both P < 0.01). Sequence analysis showed a complete consistency between the positive segment of cyclin A and the objective gene in GeneBank. (2) The levels of cyclin A protein and mRNA were positively correlated with the cumulative percentages of cells in S and G(2)/M phases (P < 0.01). (3) The expression level of cyclin A protein in the recurrent acute lymphoblastic leukemia (ALL) group was 15.4%, significantly lower than those of de novo group (29.5%, t = 14.418, P = 0.022). (4) The complete remission (CR) rates in the AL patients with high expression levels of cyclin A protein and mRNA group were 87.9% and 85.7% respectively, significantly higher than those in the AL patients with low expression levels (38.2% and 53.5% respectively, both P < 0.01). Multivariate regression analysis showed that cyclin A was one of the influencing factors of CR rate of AL patients (P < 0.05). CONCLUSION: Cyclin A expression contributes to the high proliferative activity in leukemia cells. The abnormal expression of cyclin A might be a prognostic marker of CR rate in AL patients. PMID- 12887744 TI - [Treatment of erectile dysfunction by implantation of domestic made three-piece inflatable penile prosthesis, a retrospective study of 29 cases]. AB - OBJECTIVE: To evaluate the effect of implantation of domestic made three-piece inflatable penile prosthesis on erectile dysfunction (ED). METHODS: Domestic made three-piece inflatable penile prosthesis were implanted through an incision at the border of penis and scrotum on 29 patients with ED refractory to non operative treatment, among which 23 being with neurogenic ED, including 18 paraplegia cases and 5 cases of traumatic injury of nervi erigentes; 4 cases with ED caused by arteriovenous fistula; I case with ED caused by fibrosis of corpus cavernosum, and 1 case with ED caused by Klinefelter's syndrome. Follow-up was conducted for 3 - 36 months to survey the clinical effect. RESULTS: Operation was successfully performed upon all 29 patients. Follow-up after operation lasted 3 - 36 months, 18 months on average. Fluid exudation of penile prosthesis and localized infection occurred in 1 patient and mechanic complication occurred in 2 patients. Maintained sexual intercourse rate was 28/29. 3 ED patients with primary arteriovenous fistula made their spouses pregnant after treatment. CONCLUSION: With excellent covertness and at low cost, implantation of domestic made three-piece inflatable penile prosthesis is effective and well accepted by ED patients. PMID- 12887745 TI - [Targeted tumor suppression by a secreted fusion protein consisting of anti- erbB2 antibody and reversed caspase-3 to SKBr3 cells]. AB - OBJECTIVE: To investigate the targeted killing effect to SKBr3 cells due to the expression of a secreted fusion protein consisting of anti-erbB2 antibody and reversed caspase-3. METHODS: A recombinant plasmid pCMV-e23scFv-PEII-revcasp 3 was constructed by subcloning reversed caspase-3 gene to the downstream of anti erbB2 antibody and transfected into Jurkat cells. The cell lines which secreted expressing fusion protein stably were selected. The fusion protein in media was detected by ELISA and the media was used to culture human breast cancer SKBr3 cells. The recombinant plasmids with liposomes was administrated to BALB/C nude mouses bearing SKBr3 tumor by intramuscular injection. The targetting effect of the recombinant fusion protein caspase-3 was detected by indirect immunofluorescence staining. RESULTS: Fusion protein can be expressed and secreted by Jurkat cells stably and kill SKBr3 cells. Significant prolonged survival time (prolonged by 72%) and inhibition of tumor growth in vivo (within inhibition ratio of 77%) were seen in the group administered with recombinant plasmids. Indirect immunofluorescence staining showed that the recombinant fusion protein caspase-3 has targetting effect. CONCLUSION: Secreted expression of the fusion protein consisting of anti-erbB2 antibody and reversed caspase-3 can targetedly induce SKBr3 cells to death. PMID- 12887746 TI - [Transcriptional inactivation of RASSF1A in lung cancer and its clinical significance]. AB - OBJECTIVE: To explore the relationship between the transcriptional expression of RASSF1A (Ras association domain family 1A gene) and oncogenesis and development of lung cancer. METHOD: RT-PCR was used to detect the expression of RASSF1A mRNA in 47 human lung cancer tissues and matched 47 non-cancer tissues. RESULTS: (1). The RASSF1A mRNA was identified in all non-cancer tissues but not found in 53.2% carcinoma tissues. (2). The rate of loss of RASSF1A mRNA was significantly higher in the patients with positive lymph node metastasis (70.4%) than in those without lymph node metastasis (30.0%) (P < 0.01). (3). Much more frequent in advanced tumor tissues, loss of RASSF1A mRNA was correlated with TNM stage (P < 0.05). (4). No significant association of abnormal RASSF1A expression was identified with histological type, differentiation grade of tumors or age, sex, and smoking index of the patients (all P > 0.05). CONCLUSION: RASSF1A is one of the new candidates of tumor suppressor genes. Loss or abnormal down-regulation of RASSF1A mRNA is a frequent event in lung carcinogenesis, which may play an important role in the malignant progression and prognosis of lung cancer. PMID- 12887747 TI - [Proliferation-inhibiting effect of advanced glycation end products modified human serum albumin to vascular endothelial cell ECV304]. AB - OBJECTIVE: To study the proliferation-inhibiting and apoptosis-inducing effects of advanced glycation end products (AGE) modified human serum albumin (AGE-HSA) on human vein endothelial cells. METHODS: Human umbilical vein endothelial cells ECV304 were cultured in vitro with AGE-HSA of the concentrations of 12.5, 25, 50, 100, and 200 micro g/ml for 6, 12, 24, or 48 hour, then 20 micro l of 5 mg/ml MTT were added and the optical density (OD) at each time point was determined. Another ECV304 cells were cultured with AGE-HAS for 2, 4, or 8 days and then were stained with trypan blue to calculate the number of dead cells so as to calculate the proliferation-inhibiting rate. Another ECV304 cells were cultured with AGE HAS for 6, 12, 24, or 48 hours and then stained with annexin V Fitc and propidium iodide (PI). Flow cytometry was used to calculate the annexin V Fitc positive cells (early and middle stage apoptotic cells) and Annexin V Fitc/PL positive cells (late apoptotic cells). Inverted microscope, transmission electron microscope, and fluorescence microscope were used to observe the histological changes of apoptotic cells. FCV304 cells incubated with HSA of the above mentioned and without addition of the other agents concentrations were used as controls. RESULTS: The OD values of ECV304 cells cultured for 48 h with low concentrations (12.5, 25, and 50 micro g/ml) of AGE-HSA were not significantly different from those of the control (1.104 +/- 0.080, 1.098 +/- 0.097 and 1.059 +/- 0.122 VS. 1.159 +/- 0.088, all P > 0.05). The OD values of ECV304 cells cultured with low concentrations of AGE-HSA for 4 days and 6 days were significantly lower than those in the control group. The OD values of ECV304 cells cultured with high concentrations (100 and 200 micro g/ml) of AGE-HSA for 6 - 48 hours decreased to 0.117 +/- 0.033 and 0.081 +/- 0.020 in comparison with that of the control group (P < 0.01). Flow cytometry and fluorescence microscopy showed higher proportions of apoptotic cells among the ECV304 cells cultured with high concentrations of AGE-HAS than among the control cells at each time point (P < 0.01). The numbers of cells in the control group exponentially increased after culture for 2, 4, and 6 days. The number of cells cultured with low concentrations of AGE-HAS for 2 days was not significantly different from that of the control group (P > 0.05), however, the numbers of cells cultured with low concentrations of AGE-HAS for 4 and 6 days were significantly lower than those of the control group (both P < 0.01). The numbers of cells cultured with 100 or 200 micro g/ml AGE-HAS for 2 days were significantly lower than those of the control group (both P < 0.01) with a proliferation-inhibiting rate of 39.56% +/- 2.82% and 60.32% +/- 4.51% respectively. The apoptotic rates in cells cultured with low concentrations of AGE-HAS for 48 hours were not significantly different from those in the control group. The apoptotic rates in cells cultured with 100 or 200 micro g/ml AGE-HAS for 6, 12, 24, or 48 hours were significantly higher than those in the control group (all P < 0.01). The apoptotic rates in 200 micro g/ml group at different time points were significantly higher than those in the 100 micro g/ml group (P < 0.05 or 0.01). The apoptotic rate and number of apoptotic cells increased along with the increase of culture time and concentration of AGE HAS. Microscopy showed morphological changes among the cells cultured with 100 micro g/ml AGE-HAS for 6, 12, 24, and 48 hours and the numbers of apoptotic cells, mainly late apoptotic cells, and dead cells increased remarkably since the cells were cultured for 48 hours. CONCLUSION: AGE-HSA inhibits the proliferation of vascular endothelial cells and induces apoptosis in dose and time dependent manner. AGE modification-induced pathobiological cascade may be involved in the pathogenesis of impaired wound healing in diabetes by the mechanism of angiogenesis retardation. PMID- 12887748 TI - [The experimental study of tissue engineered autologous cartilage using chitosan gelatin complex scaffolds]. AB - OBJECTIVE: To investigate whether man-made porous chitosan-gelatin complex scaffold was a appropriate scaffold for tissue engineering cartilage. METHODS: Chondrocytes isolated from Changfeng crossbred swines' auricular cartilage were seeded onto chitosan- gelatin scaffolds to be cultured in a three dimensional environment. The chondrocyte- polymer constructs were implanted into the subcutaneous tissue of the swines' abdomenal wall. Specimens were harvested and analyzed by gross observation, histology, type II collagen immunohistochemistry and biochemistry after 10 and 16 weeks in vivo respectively. RESULTS: H.E staining showed cartilage was formed, and chondrocytes were enclosed in lacuna with histological characteristics similar to natural cartilage. Some clusters of neocartilage surrounded by fibrous tissues were observed. Elastic fibres were observed in the mesenchyma of cartilage 16 weeks after by Vehoeff's staining. Immunohistochemical staining of the neocartilage with anti- type II collagen showed the presence of type II collagen in the ECM of tissue engineered cartilage. The proteoglycans content in tissue engineered cartilage was close to that of natural swine's auricular cartilage. CONCLUSION: The experiments demonstrated that using chitosan-gelatin complex scaffold we can generate autologous cartilage on animals with normal immune system. Porous chitosan- gelatin complex scaffolds may be a suitable scaffolds for tissue engineered cartilage. PMID- 12887749 TI - [Effects of collagen I coating on the porous poly-lactide-co-glycolid on adhesion, proliferation, and differentiation of mesenchymal stem cells]. AB - OBJECTIVE: To investigate the effects of collagen I on the adhesion, proliferation, and differentiation of MSCs on PLGA. METHODS: Collagen I was added onto the surface of pores in pieces of 3-D porous poly-lactide-co-glycolid (PLGA). Bone marrow-derived mesenchymal stem cells (MSCs) were obtained from New Zealand rabbits and were cultured for 3 generations, inoculated into the pores of PLGA pieces with the volume of 0.3 cm x 1.2 cm x 2.0 cm, and then cultured in solution with [(3)H]-thymidine deoxyribose (TdR). PLGA pieces not coated by collagen I were used as controls. The incorporation rate of [(3)H]-TdR was detected 2, 4, 6, and 8 hours, and 7, 14, and 21 days after culture, shown in count per minute (CPM) value, to determine the adhesion and proliferation of the MSCs. RT-POCR was used to examine the expressions of mRNA of the osteoblast markers: osteocalcin (OCN), alkaline phosphatase (ALP), and osteopontin (OPN). Scanning electron microscopy (SEM) was used to observe the morphology of MSCs. RESULTS: The CPM value since 6 hours after culture between the experimental group and control group began to be significantly different (both P < 0.05) The CPM values 7, 14, and 21 days after culture between the experimental group and control groups (P < 0.05 or P < 0.01). OCN, ALP, and OPN mRNA were expressed in MSCs of the experimental group and only ALP mRNA was weakly expressed in the control group. SEM showed the distribution of spindle and polygonal cells in the pores of the 3-D PLGA pieces and distribution of cylindrical or round cells in the control group. CONCLUSION: Collagen I is effective in promoting the adhesion, proliferation, and differentiation of MSCs on PLGA. PMID- 12887750 TI - [Immune tolerance inducing effects of soluble human leucocyte antigen G1 on natural killer cells and T cells]. AB - OBJECTIVE: To investigate the immune tolerance inducing effects of soluble human leucocyte antigen G1 (sHLA-G1) on natural killer (NK) cells and T cells. METHODS: A recombinant plasmid expressing sHLA-G1 was constructed and transfected into human lymphoblastoid cells LCL721.221. sHLA-G1 in the supernatant was purified by immuno-affinity chromatography and then added into the culture of NK cells obtained from the peripheral blood mononuclear cells of 3 unrelated individuals. Target cells, K562 cells, were added too. The killing rate of NK was calculated. Peripheral blood lymphocytes (PBLs) were obtained and stimulated by Ebstein-Barr virus-transformed B lymphoblastoid cell line (EBV-LCL). The proliferation of the T cells in the mixed lymphocyte culture was examined by enzyme linked immunosorbent assay. The antigen-specific T cells in the peripheral blood was activated. sHLA-G1 was added into the culture. Then the T cells were suspended in the solution of fluorescence isothiocyanate (FITC)-annexin-V. Flow cytometry was used to detect the fluorescent intensity of FITC so as to examine the apoptosis of T cells. RESULTS: sHLAG-1 inhibited the cytotoxicity of NK cells dose dependently. sHLAG-1 inhibited the proliferation of activated T cells, and induced the apoptosis of T cells dose-dependently, with a dose-saturation character and without antigen-specificity. CONCLUSION: sHLAG-1 is a kind of immune tolerance inducing molecule. PMID- 12887751 TI - [Early expression of pro-inflammatory cytokines in the lung of rat with small-for size liver transplantation and the significance]. AB - OBJECTIVE: To investigate the expression of tumor necrosis factor-alpha(TNF alpha), interleukin-1 beta (IL-1beta), and intercellular adhesion molecule-1 (ICAM-1) in the lung of rat with small-for-size liver transplantation and the significance of the expression of these cytokine in lung injury after liver transplantation. METHODS: 150 SD rats were randomly divided into 4 groups: sham operation group (n = 6), whole graft liver transplantation group (n = 48), 50% size graft liver transplantation group (n = 48), and 30% size liver transplantation group (n = 48). Six rats from each group were killed 0.5, 2, 6, and 24 hours after operation. Blood samples from subhepatic inferior vena cava were obtained to examine the plasma TNF-alpha by ELISA. Specimens of lung were obtained to be examined pathologically. RT-PCR was used to examine the expression of TNF-alpha mRNA, IL-1beta mRNA, and ICAM-1 mRNA in lung, and chromatometry was performed to detect the activity of myeloperoxidase (MPO). RESULTS: (1) The plasma TNF-alpha at any time point was higher in the 3 transplantation groups than in the sham operation group. The plasma TNF-alpha 2 hours after operation in the whole graft group was significantly higher than that in the 30% size group (P < 0.05). (2) The expression levels of TNF-alpha mRNA 2 and 6 hours after operation in the whole graft group and in the 50% graft group were significantly higher than those in the 30% graft group (all P < 0.01). The expression levels of TNF-alpha mRNA remained significantly higher than those in the sham operation group since the second hour after operation (all P < 0.01). (3) IL-1beta mRNA was expressed in the 3 liver transplantation groups without significant differences between any levels at all the time points and was not expressed in the sham operation group. The expression of ICAM-1 mRNA was higher at all the time points in the liver transplantation groups than in the sham operation group (P < 0.01 or P < 0.05) without significant difference between the values of any transplantation group at any time point (all P > 0.05). The MPO activity was stronger in the 3 liver transplantation groups at any time point than in the sham operation group (all P < 0.01). The peak occurred 2 hours after operation in the whole graft group and 50% size group and occurred 6 hours after operation in the 30% graft group. The plasma TNF-alpha level was positively correlated to the MPO activity in lung tissue with a correlation coefficient of 0.422 (P < 0.05). (4) The morphology of lung was normal in the sham operation group. Obvious interstitial hyperemia and hemorrhage, neutrophil infiltration, and pulmonary septal thickening were found in the 3 transplantation groups, in particular being severe at the 2-hour time point. CONCLUSION: Increase of plasma TNF-alpha is one of the causes of lung injury after small-for-size liver transplantation. Upregulation of TNF-alpha mRNA, IL-1beta mRNA, and ICAM-1 mRNA expression may be also responsible for the lung injury and their expression may be correlated to the size of graft. PMID- 12887752 TI - [Expression of Pax3 and Cx43 in NTD embryos induced by hyperglycemia]. AB - OBJECTIVE: To find out the molecular mechanisms of togenesis, especially neural tube defects (NTDs) caused by hyperglycemia and thiadiazole. METHODS: Mice were raised with a ratio of 2:1 between females and males. Forty-five pregnant female mice were randomly divided into 5 groups: blank control group (without any treatment), high-glucose group (treated with subcutaneous injection of 25% glucose), glucose-control group (n = 12, treated with subcutaneous injection of 5% glucose), thiadiazole group (treated with gastric perfusion of thiadiazole), and thiadiazole-control group (n = 11, treated with the solvent of thiadiazole). The blood sugar was examined among the mice of the first three groups on the 8th gestational day. All of the mice were killed on the 15th gestational day and the embryos were taken. The existence of teras was observed. RT-PCR was used to detect the expression of Pax3 and of Cx43 genes and immunohistochemistry was used to examine the Pax3 and Cx43 proteins in the embryos. RESULTS: The blood sugar of the pregnant mice in the high-glucose group was 13.4 mmol/L +/- 0.8 mmol/L, significantly higher than those in the 2 control groups (4.9 mmol/L +/- 0.4 mmol/L and 4.8 mmol/L +/- 0.4 mmol/L respectively, both P < 0.01). The teras rate, absorbed embryo rate and still embryo rate in high-glucose group were significantly higher than those in the 2 control groups (all P < 0.01). The embryo weight of the high-glucose group was significantly lower. The expression of Pax3 in the hyperglycemia group was significantly lower than those in the 2 control groups (both P < 0.01), while the expression of Cx43 in high-glucose group was significantly higher than those in the 2 control groups (both P < 0.01). In the thiadiazole group, the expression of Pax3 was not significantly different from, and the expression of Cx43 was significantly higher than those in the 2 control groups. CONCLUSION: Hyperglycemia is induced in nondiabetic pregnant mice by subcutaneous glucose injection, and elevated glucose appears to be critical in the diabetic embryopathy. Hyperglycemic episodes disturb the essential embryonic control genes: decrease the expression of Pax3 gene and increase the expression of Cx43 gene, thus causing congenital defects. Thiadiazole induces NTDs by increasing the expression of Cx43. PMID- 12887753 TI - [Expression of human fibroleukin gene acute on chronic hepatitis B and its clinical significance]. AB - OBJECTIVE: To investigate the mRNA and protein expressions of human fibroleukin gene (hfg12) in acute on chronic (AOC) hepatitis B and its clinical significance. METHODS: Liver tissues were obtained from 23 patients with AOC hepatitis B, 13 patients with chronic hepatitis, and 14 patients with cirrhosis to be examined histologically. Immunohistochemistry and in situ hybridization were used to detect the mRNA and protein expressions of hfg12 in the liver tissues. Double staining was used to the hfg12 positive samples to examine both the hfg12 and fibrin. Four specimens of liver tissue from normal donors were used as controls. RESULTS: Immunohistochemistry showed that hfg12 was expressed in the liver tissues of 21 out of the 23 patients with AOC hepatitis B (91.30%) and only one out of the 13 patients with chronic hepatitis (7.69%). In situ hybridization showed that hfg12 was expressed in the liver tissues of 13 out of he 23 patients with AOC hepatitis B and in none of the 27 patients with chronic hepatitis or cirrhosis. In patients with AOC hepatitis Kupffer's cell, CD68 positive, was numerous and big, mainly distributed in the necrosis areas. It was identified as the same of hfg12-expressing cells. CONCLUSION: High expression of hfg12 is one of the molecular mechanisms of necrosis of liver cells in AOC hepatitis. PMID- 12887754 TI - [The analysis of S gene phylogenetic tree of HBV in transmission from father to infant]. AB - OBJECTIVE: To study HBV transmission from father to infants, the study was carried out. METHODS: The study contained 16 pairs of fathers who was HBV carriers, and infants whose mothers wasn't HBV carriers. The infants infected HBV in womb. The homogenous of HBV S gene were compared between fathers and infants, meanwhile HBV gene phylogenetic tree were analyzed. RESULTS: The genotype of 16 pairs fathers and infants were HBV adw. The homogenous of HBV S gene were 98% approximately 100%. The mutation of 488, 491, 494, 530, 531, 546, 581, 621 nucleotide of HBV S gene caused in 112, 113, 114, 126, 131, 143, 156 amino acid substitution. The mutation of 126 amino acid from Threonine to Alanine didn't existed in the gene bank. The sequence was difference from gene classification. CONCLUSION: The homogenous of HBV S gene of fathers and infants was very high. The HBV gene characteristic in China was difference from gene bank. PMID- 12887755 TI - [The changes and significance of red cell nature-immune-adhesion function in liver diseases at different stages]. AB - OBJECTIVE: To study the changes of red cells nature immune adhesion function (RNIAF) in liver diseases at different stages, and the feasibility of RNIAF in evaluating the severity of liver diseases. METHODS: Venous blood was extracted from 682 patients with liver disease, including cirrhosis, chronic hepatitis, acute hepatitis, hepatitis gravis, and 50 healthy blood donors as controls. Suspension of red cells in self-plasma was mixed with solution of mouse ascites carcinoma cells. A tumor cell attached with 5 red cells or 2 lymphocytes/granulocytes was counted as one rosette. The adhesion rate was calculated. Serum soluble complement receptor 1 (sCR1) was measured with a newly established sandwich enzyme-linked immunosorbent assay (ELISA). The CR1 expressed on erythrocytes has assayed by cell-ELISA. RESULTS: The expression of CR1 on erythrocytes was lower among the patients with liver diseases. Compared with that in healthy controls, RNIAF decreased among the patients with various liver diseases (all P < 0.01) in the order of acute hepatitis, chronic hepatitis, cirrhosis Child's A, cirrhosis Child's B, cirrhosis Child's C, and hepatitis gravis. CHE and PTA decreased and PT increased among the patients with cirrhosis and hepatitis gravis (all P < 0.05 or P < 0.01). RNIAF was positively correlated with CHE, PT, and PTA in patents with cirrhosis. CHE, PT and PTA remained at low levels while RNIAF almost returned to its normal level in the convalescents of hepatitis gravis. CR1 expressed on erythrocytes and sCR1 in serum in various liver diseases were decreased at different degrees, however, they changed significantly later than RNIAF. Serum sCR1 significantly increased in the patients with cirrhosis, particularly in those in grades Child's A to C. CONCLUSION: Low-cost, easy to operate and with early change and stable results, RNIAF may be used as an important indicator to evaluate the severity of liver diseases. of CHE, PT/PTA. It is an important to evaluate the severity of liver function in clinic. PMID- 12887756 TI - [Study of gene chips in the detection of YMDD mutations in the region of HBV polymeras]. AB - OBJECTIVE: To understand mutations of the HBV polymerase from the level of molecule-biology and to guide reasonably antiviral therapy in clinic and predict prognosis of the patients, the gene chips were applied to detect YMDD mutations for the serum samples with HBV DNA position before antiviral therapy. METHODS: The serum samples from 150 HBV DNA positive patients non-antiviral therapy were detected by nowly advanced gene chip technique for YMDD wildtype and YVDD and/or YIDD mutations in the region of HBV polymerase. Several positive samples had been testified by gene sequences. RESULTS: Out of 150 serum samples of HBVDNA position patients tested by gene chips, 122 cases (including YMDD wildtype and mutations) were positive (81.3%), 28 cases were negative (18.7%). In 122 positive samples, 90 cases were YMDD wildtype (73.8%) (non-mutations), 28 cases were YVDD mutation (22.9%), 2 cases were YIDD mutation (1.6%), 2 cases were YVDD/YIDD double mutations (1.6%). Total mutation rate is 26.2%. Sequence analysis of 8 positive samples verified that gene chips were available. CONCLUSION: (1) Gene chip technique for the detection of mutations of the YMDD in the region of the HBV polymerase is reliable and its specificity is high. (2) Gene chips can simultaneously and quickly detect HBV YMDD wildtype and YVDD, YIDD mutations. (3) HBV YMDD mutations have existed in several patients HBV DNA position before antiviral therapy and YVDD mutation is high. YMDD wildtype and mutation strain coexist. (4) To guide reasonably antiviral therapy in clinic and make therapeutic remedy, it is necessary for HBV-infected patients before or after antiviral therapy to detect HBV YMDD mutations. PMID- 12887757 TI - [Correlation between polymorphism of TaqI of ET-1 gene and cirrhotic portal hypertension]. AB - OBJECTIVE: To investigate the correlation between polymorphism of TaqI of endothelin (ET)-1 gene intron 4 and cirrhotic portal hypertension and to search new risk factor of portal hypertension. METHODS: Peripheral venous blood was extracted from 106 patients with cirrhosis after hepatitis B (PH+ group) and 108 healthy blood donors (PH- group). PCR-RFLP was used to analyze the polymorphism of TaqI of ET-1 gene. The plasma ET-1 concentration was detected with immunoassay. Multivariate logistic regression analysis was made to analyze the risk factors. RESULTS: The C allele frequency in the PH+ group was 25.4%, significantly higher than that of the controls (16.7%, P < 0.05). The frequency of CC + TC genotype in PH+ group was 46.2%, significantly lower than that in the controls (29.6%, P < 0.05). In the PH+ group, the thickness of spleen was greater, hemorrhage rate was higher, and III degrees ascites was more in C allele carrier than in T allele carriers (P < 0.05). The plasma ET-1 concentration was higher in PH+ group than in PH- group. In the PH+ group, the plasma ET-1 concentrations in those with CC genotype and those with TC genotype were significantly higher than in those with TT genotype (P < 0.05). Correlation analysis showed that ET-1 gene polymorphism was positively correlated with plasma ET-1 concentration (R = 0.808 2). Multivariate logistic regression analysis showed that gradation of liver function, diameter of portal vein, and ET-1 gene polymorphism were independent risk factors of portal hypertension. CONCLUSION: Polymorphism of TaqI of ET-1 gene is correlated with the pathogenesis of cirrhotic portal hypertension. It may be one of the risk factors of portal hypertension. PMID- 12887758 TI - [Typing and prognosis of striatocapsular hemorrhage, study of 181 cases]. AB - OBJECTIVE: To analyze the impact of hematoma location and vascular territory on the clinical symptoms and signs, neuroimaging findings, and clinical prognosis of striatocapsular hemorrhage. METHODS: The data of 181 cases of striatocapsular hemorrhage, 124 males and 57 females, aged 28 - 89, including medical history, symptoms, and signs were analyzed. On the basis of hematoma locations and arterial territories, the striatocapsular hemorrhage was divided into six types: anterior type (with the blood supply from Heubner's artery), middle type (with the blood supply from medial lenticulostriate artery), posteromedial type (with the blood supply from anterior choroidal artery), posterolateral type (with the blood supply from posteromedial branches of lateral lenticulostriate artery), lateral type (with the blood supply from the most of lateral branches of lenticulostriate artery), and massive type (with the hematoma occupying the whole or greater part of striatocapsular region). The short-term clinical outcome and prognosis were evaluated by Modified Rankin Scale (MRS). The relationship between the type and outcome was analyzed. RESULTS: There were 22 cases of anterior type, accounting for 12.15%, of which 68.18% showed an excellent outcome (MRS = 0 - 2) and only 1 case died. There were 12 cases of middle type, accounting for 6.63%, of which 6 cases (50%) showed a poor outcome (MRS = 5) but without dead case. There were 19 cases of posteromedial type (10.50%), of which 52.63% had excellent outcome (MRS = 0 - 2) without dead case. There were 19 cases of posterolateral type (18.78%), of which 76.47% were with poor outcome (MRS = 5) and 17.65% died. Accounting for 39.23%, lateral type was the most common condition, of which 38.03% patients had a poor outcome (MRS = 5) and 8.45% cases died. The incidence rate of massive type was also high (12.71%), of which 47.82% showed a poor outcome (MRS = 5) and 47.83% cases died. CONCLUSION: Hematoma location and vascular territory are the determinating factors of prognosis of striatocapsular hemorrhage. The six-type classification system provides a simple and reliable means to predict the clinical outcomes and to guide therapeutic options of striatocapsular hemorrhage. PMID- 12887759 TI - [Noninvasive monitoring of hypertensive intracerebral hemorrhage and perihematomal tissue edema]. AB - OBJECTIVE: To investigate the dynamic change regularity of intracerebral hemorrhage and perihematomal tissue edema. METHODS: The cerebral electric impedance (CEI) of 100 normal volunteers and 24 patients with hypertensive intracerebral hemorrhage was measured, and the results of measurement were compared with the volumes of hematoma detected by CT. RESULTS: In the normal persons, the CEI values in the left and right sides of cerebral hemispheres were 11.5 +/- 1.3 k(Omega) vs 11.4 +/- 1.8 k(Omega) at the frequency of 20 kHz, 10.2 +/- 1.0 k(Omega) vs 10.3 +/- 1.1 k(Omega) at the frequency of 50 kHz, and 9.1 +/- 0.8 k(Omega) vs 9.2 +/- 0.8 k(Omega) at the frequency of 100 kHz (all P > 0.05). Age, sex and measuring time did not obviously affect the CEI values. In the patients with hypertensive intracerebral hemorrhage the CEI of hematoma side was significantly decreased within 2 days after onset and then increased significantly for more than 10 days. The rate of EI change in the hematoma side within 24 h had a negative correlation with the volume of hematoma (r = -0.895 3, P < 0.01). however, the rate of CEI change in the hematoma side had a positive correlation with the volume of perihematomal edema (r = 0.881 1, P < 0.01). CONCLUSION: CEI is a stable physical parameter reflecting the electrical character of human brain tissue. CEI sensitively reflects the change of hematoma and perihematomal edema, and guides the clinical treatment of cerebral hemorrhage. PMID- 12887760 TI - [Effect of movement arousal and hypoxia on circardian blood pressure in hypertensive patients with obstructive sleep apnea syndrome]. AB - OBJECTIVE: To investigate whether repeated arousals and hypoxia at the end of apnea contribute to nocturnal and diurnal elevation of blood pressure in obstructive sleep apnea syndrome (OSAS) patients with hypertension. METHODS: 30 OSAS patients with hypertension and 30 OSAS patients without hypertension were studied. Movement arousal index (MAI), cortical arousal index (CAI), length of time during which nocturnal oxygen saturation decreased below 90% (OLT90%), apnea/hypopnea index (AHI), 24 h average systolic and diastolic blood pressure (ASBP and ADBP) were measured. Multiple stepwise regression analysis was performed to find the correlation between 24 h ASBP, ADBP and MAI, CAI, OLT90%, AHI, age, body mass index in OSAS patients with hypertension. RESULTS: MAI made the most significant contribution to 24 h ASBP and ADBP in OSAS patients with hypertension (ASBP: r = 0.485, P < 0.01; ADBP: r = 0.407, P < 0.05). OLT90% was the second most important contributing factor (ASBP: r = 0.382, P < 0.05; ADBP: r = 0.369, P < 0.05). CONCLUSION: Movement arousal and hypoxia may make an important contribution to circardian elevation of blood pressure in OSAS patients with hypertension. Both of them may contribute to the development of nocturnal and diurnal hypertension in patients with OSAS. PMID- 12887761 TI - [Arterial switch operation in older infants with severe pulmonary hypertension]. AB - OBJECTIVE: To investigate the clinical efficacy of arterial swith operation on transposition of great artery (TGA) and Tausing-Bing anomaly. METHODS: Between June 2000 and December 2002, 30 consecutive patients, aged 3 days to 6 years (mean, 9.4 +/- 15 months) with the mean body weight was 6.1 kg +/- 2.7 kg, underwent arterial switch operation. Among the 30 patients 7 suffered from TGA with intact ventricular septum, 19 from TGA with ventricular septal defect (VSD), 3 from Taussing-Bing anomaly, and 1 from corrected TGA; 12 were complicated by atrial septal defect (ASD) and 18 complicated by patent ductus arteriosus (PDA); 23 had severe pulmonary hypertension; 2 had left ventricular outlet stenosis. Coronary type A distribution was recognized in 26 cases, type D in 4, and one of them having the origin of the left descending artery tunneled in the aortic wall. The operation was performed under general anesthesia and extracorporeal circulation with low temperature and low volume blood flow. Prostaglandin 1 was administered pre-operatively in 3 patients, one of which underwent balloon atrial septostomy and one underwent pulmonary banding and systemic to pulmonary shunt pre-operatively. The aorta and pulmonary artery were transected above the valvular commisures, the coronary ostia with all the adjacent sinus of Valsalva were excised and re-implanted to the proximal neo-aorta, and then aortic anastomosis was completed. The proximal neo-pulmonary trunk was reconstructed with a large autologous native pericardium as a posterior patch. The pulmonary anastomosis was completed after the aortic cross-clamp was released. The VSD was repaired through the atrium or proximal aorta with dacron patches. RESULTS: Two patients died with a hospital mortality rate of 6.7%. No death was directly related to any coronary artery problem. One 5 day-old neonate with TGA and an intact septum having refractory hypotension, hypoxemia, and acidosis pre operatively underwent a smooth emergency operation. The patient had a refractory low cardiac output syndrome post-operatively and died 20 hours after the operation. Another patient with chylothorax died of allergy from iodophor 22 days postoperatively. The pulmonary pressure decreases significantly in 22 patients who had severe pulmonary hypertension preoperatively with the mean pressure 46.7 mm Hg preoperatively and 31.3 mm Hg postoperatively. 28 patients were discharged from hospital uneventfully. Follow-up of 1 to 31 months showed survival with no late complications and death. CONCLUSION: The arterial switch procedure has a satisfying effect on TGA for patients older than 1 month with severe pulmonary hypertension. PMID- 12887762 TI - [Detection of cell-free fetal DNA in the urine of pregnant women by nested polymerase chain reaction]. AB - OBJECTIVE: To investigate the clinical feasibility of fetal sex determination by using nested polymerase chain reaction (PCR) to detect cell-free fetal DNA in the urine of pregnant women. METHODS: The urine specimens of 40 healthy predelivery women were centrifuged. The DNA in the specimens of supernatant was extracted by using of QIAamp Viral RNA Mini Kits. Nested PCR was used to specifically amplify the SRY gene, 300 bp in length, on Y chromosome in order to detect the fetal DNA from the women bearing male fetuses. One specimen of normal man's urine and one specimen of non-pregnant woman's urine were used as controls for each specimen from pregnant women. The results were confirmed by the sexes of neonates examined after birth. RESULTS: Amplified band of SRY was found in 6 specimens. Sex examination after delivery showed 21 male neonates and 19 female neonates. The positive rate of detection of SRY gene was 28.6% (6/21) and the false negative rate was 71.4%, and false positive rate was 0. Five out of the 6 cases with positive results showed positive 2 times and one out of the 6 cases only showed positive at the second time. CONCLUSION: There is cell-free fetal DNA in the urine of pregnant women. Nested PCR is useful in detection of fetal sex, however, still with a rather low positive rate. PMID- 12887763 TI - [Adenoviral vector expressing an antiangiogenic fragment of thrombospondin 1 inhibits the growth of K562 cell xenografts]. AB - OBJECTIVE: To explore the effect of adenovirus-mediated delivery of an antiangiogenic fragment of human thrombospondin 1 (TSP1(f)) on K562 cell growth in nude mice. METHODS: TSP1(f) cDNA was amplified by RT-PCR from normal human peripheral blood mononuclear cells and was used as transgene to construct a adenoviral vector (ADV-TSP1(f)). Human leukemia K562 cells were cultured and infected with ADV-TSP1(f) or ADV-LacZ. PBS was used as control. TSP1(f) expression/secretion by these infected K562 cells was demonstrated using Western blot analysis. MTT assay was performed to determine the effect of ADV-TSP1(f) infection on K562 cell growth kinetics. Eighteen female Balb/c nude mice were inoculated subcutaneously with human leukemia K562 cells. When the diameter of the tumor reached 5 approximately 7 mm the rats were randomly divided into 3 groups of 6 rats injected intratumorally with ADV-TSP1(f), ADV-LacZ, and PBS respectively. The volume of K562 xenografts was measured every three days during the 3-week treatment. By the end of the 21st day the mice were killed and the tumors were taken to undergo histological examination. The intratumoral microvessel density (MVD) was determined by immunohistochemical staining. RESULTS: TSP-1f was expressed and secreted efficiently by ADV-TSP1(f)-infected K562 cells. Three weeks after the initial treatment, the volume of K562 xenografts in the mice treated with ADV-TSP1(f), ADV-LacZ, and PBS was (1,108 +/- 179) mm(3), (4,518 +/- 452) mm(3), and (4,666 +/- 458) mm(3) respectively (P < 0.01). The number of CD31+ microvessels counted per x200 field was 34 +/- 9, 36 +/- 7, and 14 +/- 4 in the tumors treated with PBS, ADV.LacZ, and ADV.TSP-1f respectively (P < 0.001). CONCLUSION: Adenovirus-mediated TSP-1f gene transfer greatly inhibits K562-derived tumor growth and angiogenesis in mouse xenograft model, and may serve as a new therapy for hematological malignancies. PMID- 12887764 TI - [The Fc region of immunoglobulin suppresses atherosclerosis in apolipoprotein E knockout mice]. AB - OBJECTIVE: To investigate the role of immunoglobulin in inhibition of atherosclerosis and its mechanism. METHODS: Apolipoprotein E knockout mice aged 6 weeks were fed with high fat diet containing 20% fat and 0.3% cholesterol for 8 approximately 16 weeks to induce the formation of fatty streak and fibrofatty plaque and were injected intraperitoneally with either human intact immunoglobulin (1 g x kg(-1) x d(-1)) or F(ab')(2) fragments of human immunoglobulin (1 g x kg(-1) x d(-1)) once the other day for 8 or 16 weeks. Sibling mice were injected intraperitoneally with human serum albumin (HSA) 1 g/kg once the other day as controls. 16 weeks later, the mice were killed. Their blood in right atrium was extracted to examine the total cholesterol (TC), high density lipoprotein cholesterol (HDL-C), and triglyceride (TG). The root of aorta and ascending aorta were isolated and made into tissue slices to be examined histochemically to calculate the areas of fatty streak and fibrofatty plaque. Immunohistological staining was conducted to examine the expression of macrophages CD(4)(+) T cells and CD(8)(+) T cell and infiltration of I-A(b+) cells in the specimens of aorta roots. IG or F(ab')(2) fragments and then lipopolysaccharide (LPS) was added into the culture media of cells of human monocyte/macrophage line U(937) carrying Fcgamma receptor surface marker: CD(16), CD(32), and CD(64) molecules. Flow cytometry was used to detect the expression of these Fcgamma receptor surface molecules. RESULTS: The area of fatty streak formation in the mice treated with IG was 4. 22.0%, significantly smaller than that in the control mice (13.6% +/- 4.8%, P < 0.01). However, the area of fatty streak formation in the mice treated with F(ab')(2) fragments was 12.1% +/- 3.7%, not significantly different from that of the controls. The area of fibrofatty plaque in the mice treated with IG was 8.1% +/- 2.7%, significantly smaller than that of the control (21.5% +/- 3.9%, P < 0.01). However, the area of fibrofatty plaque in the mice treated with F(ab')(2) fragments was 20.6% +/- 4.0%, not significantly smaller than that in the controls. Immunohistochemical analysis revealed that the percentage of M(Phi) positive cells in the fatty streak lesions was lower in the mice treated with IG than in the controls, however, the percentage of M(Phi) positive cells in the fatty streak lesions in the mice treated with F(ab')(2) fragments was not significantly different from that in the controls (P = ns vs HSA). There was no significant difference in the expression of CD(4)(+) T cells, CD(8)(+) T cells and the infiltration of I-A(b+) cells between the mice treated with IG or F(ab')(2) fragments and the controls. The serum TC, HDL-C, and TG were not significantly different among any groups. The mean fluorescence intensity (MIF) of CD(32) molecule in the surface of U(937) cells after cultured with IG and LPS was 108% +/- 18%, significantly lower than that in the surface of U(937) cells after cultured with only LPS (156% +/- 26%, n = 4, P < 0.05), however, the MIF of CD(32) molecule in the surface of U(937) cells after cultured with F(ab')(2) fragments was not significantly different from that in the surface of U(937) cells after cultured with only LPS. CONCLUSION: Immunoglobulin therapy remarkably suppressed atherosclerosis due to Fc receptor-mediated anti-inflammatory action. The suppression of the disease is associated with remarkably reduction of macrophage density in the lesions, but not with the reduction of high serum lipid levels. PMID- 12887765 TI - [Molecular basis of preventive effect of human apolipoprotein-1 on murine vascular smooth muscle cell proliferation and lipid deposition induced by oxidized low density lipoprotein]. AB - OBJECTIVE: To study the molecular basis of preventive effect of human apolipoprotein-1 (h-apoA-1) on vascular smooth muscle cell (vSMC) proliferation and lipid deposition induced by oxidized low density lipoprotein (ox-LDL). METHODS: Smooth muscle cells originated from the aortae of h-apo-A-1 transgenic mice were cultured and divided into 2 groups, one group was stimulated by ox-LDL (tester) and the other group was used as control (driver). Subtractive hybridization was used to enrich the genes differentially expressed in the vSMCs induced by ox-LDL. A subtractive library was thus established and confirmed by colony hybridization in situ and dot blot analysis. 15 clones out of the 57 differentially expressed clones were randomly chosen foe sequencing and homology analysis. The whole-length cDNA library of vSMC induced by ox-LDL was established using SMART technique. RESULTS: Three expression sequence tags (EST), all correlated with immune system, were confirmed: C1-inhibitor (C1-INH), lectin, and T cell receptor beta. The whole-length cDNA library contained 1.5 x 10(6) pfu/ml primary recombinants with insertions 0.5 - 3 kb in length. CONCLUSION: The 3 EST may be involved in the mechanism of atherogenesis by ox-LDL and the mechanism of the function of h-ApoA-1 in retarding the progression of atherogenesis induced by ox-LDL. PMID- 12887766 TI - [The effect of estrogen and progestin on the expression of matrix metalloproteinases, tissue inhibitor of metalloproteinase and interleukin-1beta mRNA in synovia of OA rabbit model]. AB - OBJECTIVE: To observe the effect of different dose of estrogen or estrogen plus progestin on the mRNA expression of MMP-1, MMP-3, TIMP-1 and IL-1beta in synovia of female New Zealand White rabbit OA model. METHODS: Sixty-five female New Zealand White rabbits (3 kg +/-, 6 months old) were used in this study. Right knees of all the rabbits were immobilized for 5 weeks to get OA model. Then 5 rabbits were killed by overdose of anesthesia to guarantee the model. Sixty OA rabbits were randomly divided into 6 groups (A, B, C, D, E, F) with 10 each. Group A approximately E were ovariectomized and group F were sham ovariectomized. The rabbits were feed by estradiol valerate (E(2)V) everyday as follows. Group A: E(2)V 1.8 mg/d, group B: E(2)V 3.6 mg/d, group C: E(2)V 7.2 mg/d, group D: E(2)V 3.6 mg/d + MPA 3.6 mg/d, group E was the untreated control group, group F was the untreated sham control group. Five rabbits of each group were killed respectively in 8 and 16 weeks and synovia of the right knee were obtained on all knees. Reverse transcription- polymerase chain reaction were used to detect the mRNA expression of MMP-1, MMP-3, TIMP-1 and IL-1beta in the OA synovia to determine the effect of estrogen and progestin. RESULTS: RT-PCR shows an increase expression of mRNA of MMP-1, MMP-3 and IL-1beta in OA synovia tissue in ovariectomized rabbits. Estrogen supplement could decrease the expression, an enhancement effect could be observed with progestin. Low dose estrogen could decrease the ratio of MMP-1/TIMP-1 and MMP-3/TIMP-1, which could even be decreased by progestin. High dose estrogen could slightly increase the ratio, but still lower than the untreatment control group. CONCLUSION: Our study suggests that certain dose estrogen and perhaps appropriate estrogen and progestin ratio are much important to the normal effect of articular cartilage. Estrogen deficiency or much high estrogen level could both show a damage effect on articular cartilage. PMID- 12887767 TI - [Expression and function of apoptosis-related genes Bcl-2/Bax and Fas/Fas L in the course of stress ulcer]. AB - OBJECTIVE: To investigate the correlation between the expressions of apoptosis promoting gene Bax, apotosis-inhibiting gene Bcl-2, and Fas/FasL in gastric mucosa in the course of stress ulcer. METHODS: Fifty-six rats were exposed to water immersion-restraint stress (WRS) and then randomly divided into 7 groups of 8 rats: WRS 2 h, RS 3.5 h, and 2 h, 5 h, 8 h, 12 h, and 24 h after termination of WRS. Another 8 rats were used as controls. The stomachs of rats were taken out at the above mentioned time points to observe histologically. Terminal deoxynucleatidyl transferase mediated dUTP nick and labeling (TUNEL) technique was used to examine the apoptotic cells in the gastric mucosa. The expression of Bcl-2/Bax protein was examined by immunohistochemical assay. In situ hybridization was used to detect the mRNA expression of Bcl-2, Fas, and FasL. RESULTS: (1) Apototic cells were seen sporadically in the gastric mucosa of the control rats. The number of apoptotic cells began to increase since 2 hours after the beginning of WPS and reached its peak 5 hours after the termination of WRS. Since 8 - 12 hours after the termination of WRS the number of apoptosis began to decrease, however, still remained at a level significantly higher than the normal level (P < 0.05). (2) Fas mRNA and FasL mRNA were not expressed in the controls. Fas mRNA was detected 5 - 8 hours after the termination of WRS at the base of mucosa at a low level and disappeared 8 hours after the termination of WRS. FasL mRNA was detected since WRS 3.5h to 8 hours after the termination of WRS. (3) In normal gastric mucosa, Bcl-1 mRNA and protein were expressed in gastric gland zone at a middle level and Bax protein was expressed in the epithelial cells. In WRS rats Bax protein expression began to be detected in the epithelial cells lining the gastric pits since WRS 2 h, then increased, reached the peak 2 - 5 hours after the termination of WRS, and then gradually decreased and remained a little higher than normal level 24 hours later (P < 0.05). Bcl-2 expression began to decrease since the termination of WRS and then increased 8 - 12 hours after the termination of WRS and reached the normal level 24 hours later (P > 0.05). Correlation analysis showed that the expression of Bax protein was correlated with the quantity of apotosis in gastric mucosa. CONCLUSION: The alterations of Bcl and Bax may be responsible for the development of mucosa lesion and recovery. The expression of Fas/FasL has no effect on the gastric mucosa apoptosis related to WRS. Overexpression of Bcl-2 may inhibit the Fas/FasL induced apoptosis. PMID- 12887768 TI - [Renoprotective effects of nonselective endothelin receptor antagonist bosentan on rats with interstitial fibrosis following unilateral ureteral obstruction]. AB - OBJECTIVE: To investigate the effects of nonselective endothelin receptor antagonist bosentan on the progression of renal interstitial fibrosis following unilateral ureteral obstruction (UUO). METHODS: Eighteen female SD rats were randomly divided into 3 groups: intervention group (administered with bosentan for 7 days after left ureter ligation), animal model group (administered with solution of Arabic gum for 7 days left ureter ligation) and sham operation group (administered with Arabic gum for 7 days after sham operation). Seven days after the operation the rats were killed and their left kidneys were harvested. The mRNA expressions of preproendothelin-1 (prepro ET-1), type I collagen (ColI), transforming growth factor beta1 (TGF-beta1), tissue inhibitors of metalloproteinase-1 (TIMP-1) and type 1 plasminogen activator inhibitor (PAI-1) were detected semiquantitatively with reverse transcription-polymerase chain reaction (RT-PCR). The protein expressions of endothelin-1 (ET-1), ColI, TGF beta1, TIMP-1 and PAI-1were determined semiquantitatively by immunohistochemical staining assay. RESULTS: The mRNA expressions of prepro ET-1, ColI, TGF-beta1, TIMP-1 and PAI-1 were significantly upregulated in obstructed renal tissue compared to those in sham operation group (all P < 0.05). The positive staining areas of ET-1, ColI, TGF-beta1, TIMP-1 and PAI-1 in tubulointerstitium were markedly enhanced in the animal model group in comparison with those in the sham operation group (all P < 0.05). The mRNA expressions of ColI, TGF-beta1 and TIMP 1 in renal tissue subjected to ureter ligation were significantly lower in the bosentan group then in the animal model group (all P < 0.05). The positive staining areas of ColI, TGF-beta1 and TIMP-1 in the tubulointerstitium were significantly smaller in bosentan group than in the animal model group (all P < 0.05). However, there was no significant difference in the mRNA expression of PAI 1 and in the positive staining area between the animal model group and bosentan group (both P > 0.05). CONCLUSION: ET-1 may be involved in the progression of renal interstitial fibrosis following unilateral ureter ligation and blockage of its receptors with bosentan attenuates the fibroticlesion to a certain extent by possibly inhibiting TGF-beta1 and TIMP-1. PMID- 12887769 TI - [Stimulation of neuropeptide Y on heat shock protein expression in renal vascular smooth muscle and the inhibition thereon by losartan]. AB - OBJECTIVE: To study the effect of neuropeptide Y (NPY) on the expression of heat shock protein 70 (HSP70) in vascular smooth muscle cells (VSMCs) of renal arteries. METHODS: The renal veins of SD rats were isolated and broken into pieces. The VSMCs were cultured and then divided into 3 groups, neuropeptide Y (NPY), NPY + losartan, and serum-free DMEM were added into the culture respectively. Automated MTT colorimetric microassay and quantitative immunocytochemistry were used to detect the expression of heat shock protein (HSP70) in the proliferating VSMCs. RESULTS: The fluorescence intensity of labeled HPS70 in the NPY group was 1,825.10 +/- 115.55, significantly stronger than that in the control group (1595.83 +/- 186.54, P < 0.05) and the fluorescence intensity of labeled HPS70 in the NPY + losartan group (1 658.54 +/- 183.78) was not significantly different from that in the control group (P > 0.05). The MTT-OD in NPY group was 0.2626 +/- 0.0025, significantly higher than that in the control group (0.2239 +/- 0.0010, P < 0.01). and the MTT-OD in NPY + losartan group was 0.2440 +/- 0.0013, significantly lower than that in the control group (P < 0.05). CONCLUSION: NPY stimulates the proliferation of renal VSMC, promotes the expression of HPS70, and may cause hypertension. Losartan reduces the NPY stimulation over VSMC proliferation and relevant expression of intracellular HSP70. PMID- 12887770 TI - [Surgical treatment of hepatocellular carcinoma with tumor thrombus in the inferior vena cava]. AB - OBJECTIVE: To clarify the proper surgical procedure of treating hepatocellular carcinoma (HCC) with a tumor thrombus in the inferior vena cava (IVC). METHODS: Four patients with HCC with a tumor thrombus in the IVC underwent hepatectomy and thrombectomy. Following hepatectomy, tumor thrombus was removed by incising the wall of the IVC in 3 patients and from the hepatic vein in one patient. The 3 patients underwent thrombectomy under either hepatic vascular exclusion (HVE) or Satinsky's vascular clamping. RESULTS: All the operations were successful without operative death and major complications. The postoperative course was uneventful in 3 patients and pleural effusion occurred in one patient who needed thoracentesis. Follow-up showed 3 patients died after 30, 10 and 14 months, respectively, and one patient is alive for 7 months. CONCLUSIONS: HCC with tumor thrombus in the IVC is operable and the proper procedure is hepatectomy plus thrombectomy. PMID- 12887771 TI - [Surgical intervention of hepatocellular carcinoma with bile duct thrombi]. AB - OBJECTIVES: To summarize the experience of surgical intervention for hepatocellular carcinoma (HCC) with bile duct thrombi (BDT), and to evaluate the influence on prognosis. METHODS: From 1994 to 2002, 15 patients with HCC and BDT who underwent surgical intervention were retrospectively analyzed. RESULTS: The operative procedures included hepatectomy with removal of BDT (n = 7), hepatectomy combined with extrahepatic bile duct resection (n = 4), thrombectomy through choledochotomy (n = 3), and piggy-back orthotopic liver transplantation (n = 1). The 1- and 3-year survival rates were 73.3% and 40%, respectively. Two patients survived over 5 year. The survival rate of patients with portal vein invasion was significantly lower than that of patients without portal vein invasion (P < 0.05). CONCLUSIONS: Surgical intervention was effective for patients with HCC and BDT. Operation after recurrence can prolong the survival time. Liver transplantation is a new operative procedure worthy of study. PMID- 12887772 TI - [Massive hemorrhage in hepatectomy: causes and management]. AB - OBJECTIVE: To analyse the causes and the management of massive hemorrhage in hepatectomy. METHODS: With over 1 000 ml of bleeding, 4 368 patients with hepatectomy between 1955 and 2000 were analysed retrospectively. RESULTS: Among 4 368 patients receiving hepatectomy, 286 (6.5%) had massive hemorrhage because of damage to the major hepatic veins, portal hypertension, hepatic insufficiency, and the extensive adhesion around the tumor. Massive hemorrhage was managed by repair and transfixation of the damaged vessels; transfixation or devascularization of variceal bleeding; complete vessels ligation of the hepatic section with mattress suture; resection of the ruptured tumor after temporary occlusion of the porta hepatis; fibrinogen infusion; hot saline compression of the surface of the wound and/or daub biological glue; argon beam coagulation and packs placement. CONCLUSIONS: Light performance and nonforce dragging of liver can reduce massive hemorrhage caused by major vessel injury or tumor rupture. Normothetic occlusion of porta hepatis can reduce blood loss effectively when liver resection. In situ hepatectomy must be adopted if there is extensive adhesion around the tumor. Packs placement is still an effective measure to stop bleeding caused by defective coagulation and extensive blood oozing of wound surface. PMID- 12887773 TI - [Expression of 5-hydroxytamine receptors in hepatic stellate cell and action of 5 hydroxytamine on biological characteristics of hepatic stellate cell]. AB - OBJECTIVE: To investigate the expression of 5-hydroxytamine receptors in hepatic stellate cells HSCs and action of 5-hydroxytamine on biological characteristics of HSC. METHODS: Liver ex vivo perfusion of collagenase and density gradient centrifugation were used to isolate hepatic stellate cell. RT-PCR was used to detect the expression of 5-hydroxytamine receptor subtypes 1A, 2A, 2B and 3. Western blot hybridization was used to elucidate the effect of 5-hydroxytamine and its 2A receptor antagonist ketanserin and 3 receptor antagonist ondanosetron on expression of transforming growth factor-beta1 (TGF-beta1) and Smad4 in HSC. HSCs were cultured on silicone membrane. The effect of 5-hydroxytamine, ketanserin and ondanosetron on cell contraction were studied. RESULTS: HSC expressed 5-hydroxytamine receptors subtypes 1A, 2A and 2B. 5-hydroxytamine significantly increased the expression of TGF-beta1 and Smad4 in HSC (P < 0.05). This was antagonized by ketanserin, not by ondanosetron. 5-hydroxytamine induced cell contraction in a dose-dependant manner. Ketanserin antagonized this action, but ondanosetron did not. CONCLUSIONS: HSCs express 5-hydroxytamine receptors. 5 hydroxytamine could affect the biological characteristics of HSC through its receptor mediation, and may play a role in the pathogenesis of liver cirrhosis and portal hypertension. PMID- 12887774 TI - [Differential gene expression of liver carcinoma cell after transfection of by ARL-1 with microarray]. AB - OBJECTIVE: To study the difference of the gene expression profile and to identify the different expression after transfection of the ARL-1 gene. METHODS: The cDNA probes were synthesized from total RNA of study group and control group, which was differentially hybridized to cDNA chips and confirmed by a gene specific semiquantitative reverse transcription polymerase chain reaction (RT-PCR). RESULTS: Six kinds of gene expression were increased and 9 kinds of gene expression were decreased. The findings were correlated with protein metabolism, signal pathway, metastasis, and drug resistance. CONCLUSIONS: cDNA chips showed that gene expression profile of liver carcinoma cell was changed after transfection of the ARL-1 gene. It is a useful method in understanding the mechanism of drug resistance. PMID- 12887775 TI - [Clinical analysis of primary squamous cell carcinoma of the breast]. AB - OBJECTIVE: To study the clinicopathological features of primary squamous cell carcinoma (SCC) of breast and the diagnosis and treatment of the disease. METHODS: The clinical, operative and pathological data from 7 cases of SCC of breast were retrospectively analysed. RESULTS: Five patients complained of painless mass and two of them were accompanied with local pain. All patients were diagnosed as having SCC by fine-needle aspiration before operation. Two patients underwent modified radical mastectomy, and the others received radical mastectomy. Pathological examination showed primary squamous cell carcinoma of the breast with metastasis to axlliary lymph nodes in one patient. Follow-up of four patients revealed that two patients are living well and two patients died. Three patients lost to follow up. CONCLUSION: The diagnosis of primary squamous cell carcinoma of the breast is dependent on pathological results. Radical mastectomy including modified radical mastectomy is most effective in the treatment of mammary SCC. Postoperative chemotherapy and radiation are necessary auxiliary therapy. PMID- 12887776 TI - [BRCA1 protein expression in sporadic breast and its clinical significance]. AB - OBJECTIVE: To study BRCA1 gene expression in sporadic breast cancer in Han nationality and its relations to clinical pathological parameters. METHODS: Immunohistochemical method was used to detect the BRCA1 protein in paraffin embedded breast cancer tissue of 51 patients. The relationship between BRCA1 protein and clinical pathological features was analysed. RESULTS: BRCA1 protein was located in the nuclear of breast cancer cells. The rate of its down regulation of expression was 49.0% (25/51), which correlated reversely with c erbB-2 protein, but did not correlate with histological grading, tumor size, lymph node involvement and patient age. CONCLUSION: The detection of BRCA1 protein expression may be a reflection of its gene alteration. So it may be a primary method for screening the susceptibility to breast cancer. PMID- 12887777 TI - [Arthroscopic double-bundle posterior cruciate ligament reconstruction using quadriceps tendon-patellar bone autograft]. AB - OBJECTIVE: To describe an alternative procedure of arthroscopic posterior cruciate ligament reconstruction using quadriceps tendon-patellar bone autograft and evaluate the early stage follow-up results. METHODS: Twenty consecutive cases of isolated and combined posterior cruciate ligament deficiency were studied. Quadriceps tendon with patellar bone block was used for preparation of Y-shaped autograft. Double femoral tunnels and single tibial tunnel were precisely drilled respectively at their anatomical footprint. By using this technique, the anterolateral bundle was fixed and tensioned in 90 degrees flexion, whereas the postero-medial bundle was fixed in full extension. In cases of combined injury, reconstruction or primary repair was conducted in one-stage or two-stage procedure. RESULTS: All cases were followed up for 6 months to 12 months, averaging 7.5 months. Lysholm rating scales and IKDC evaluation were used to make clinical subjective evaluations. Anteroposterior translation was measured with KT 1000 arthrometer and stress view radiography. Final Lysholm evaluation showed 5 patients (25%) were excellent, 14 (70%) good, and 1 (5%) poor. IKDC evaluation showed that 1 patient (5%) was graded as A, 16 (80%) as B, 2 (10%) as C and 1 (5%) as D. Stress view examination and KT-1000 arthrometer measurement showed that the average posterior translation improved from 11.5 mm and 11.0 mm preoperatively to 5.2 mm and 5.5 mm postoperatively. One patient was graded as failure. CONCLUSIONS: Arthrosopy assisted procedure of double-bundle posterior cruciate ligament reconstruction is effective and reproducible. The short follow up clinical results are encouraging. Quadriceps tendon is an ideal autograft alternative for posterior cruciate ligament reconstruction.Combined posterior cruciate ligament injury, especially posterolateral complex insufficiency, should be early recognized and treated properly. PMID- 12887778 TI - [Effect of protein kinase on endothelial cytoskeleton induced by septic shock]. AB - OBJECTIVE: To study the effect of cGMP-dependent protein kinase (PKG) on the pathogenesis of septic shock. METHODS: Confluent endothelial cells were disintegrated and centrifugated to obtain cell lysates after being treated with LPS or PKG activator 8-Br-cGMP. PKG activity of lysates was measured with radioactive isotope label method in a reaction system of phosphorylation of specific substrate H2B by PKG, and the shape and the distribution of intracellular filamentous actin were detected by specific fluorescence staining. For the control study, the PKG specific inhibitor KT5823 was used to pretreat the endothelial cells before the administration of LPS or PKG activator 8-Br-cGMP. RESULTS: Exposure to LPS for 5, 10, 30 and 60 minutes led to a rapid time dependent increase in endothelial PKG activity (P < 0.01 compared to the blank) and the polar distribution of intracellular filamentous actin and preincubation with KT5823 abolished these effects. 8-Br-cGMP was similar to LPS. CONCLUSIONS: The results suggested that LPS can mediate PKG activation and the stress variety of filamentous actin in the vascular endothelial cells, which probably induce the endothelial hyperpermeability after septic shock. PMID- 12887779 TI - [Endovascular stent-graft repair of aortic arch aneurysms: report of 46 patients]. AB - OBJECTIVE: To evaluate the basic procedure of endovascular stent-graft repair in the treatment of aortic arch aneurysm. METHODS: >From March 2000-February 2002, a total of 46 patients with aortic arch aneurysms were treated with the custom-made endovascular stent-graft. Of them, twenty-three patients (50%) had aneurysms at the middle of the arch, 22 patients (48%) had aneurysms at the arch-descending aorta, and 1 patient (2%) had aneurysm at the descending thoracic aorta. The diameter of all stents was 0.15 - 0.25 times larger than that of the aorta proximal to the entry tear of dissection or the opening of aneurysm. The diameter of the proximal end of the stents was ranged from 34 - 38 mm. The length of stents ranged from 90 - 120 mm. The stent was made of shape memory nitinol. RESULTS: The stent was delivered successfully in 45 patients (98%). None of the patients had any access-related complications. Either the primary entry tear of aortic dissection or the inlet of aneurysm was occluded in 43 patients (96%), with an early endoleak in 2 patients (4%). All truth lumen of the dissection recovered to normal. Of the patients in acute period, 1 was referred to surgical repair and 2 died. Follow-up for 1 month to 23 months, showed late endoleak in 3 patients (7%). Forty-three patients restored normal life. CONCLUSIONS: Endovascular stent-graft could be applied in the repair of aortic arch aneurysm. Further studies are needed to to assess the long-term efficacy of this method. PMID- 12887780 TI - [Replacement of artificial tracheal prosthesis by two-stage procedure with memory alloy mesh]. AB - OBJECTIVE: To design an artificial trachea which can totally heal with the native trachea. METHODS: Using memory-alloy mesh as the skeleton to construct an artificial trachea by two-stage operation. After 2-year animal experiment we successfully performed the operation in a patient with recurrent carcinoid of the trachea, radically resected the tumor and primarily reconstructed the trachea. RESULTS: The inner side of this "sandwich" artificial tracheal prosthesis was coated with skin and outside the memory-alloy mesh was muscle and vessel pedicle with good blood supply. The upper and lower anastomosis completely healed with recipient's trachea with a full recovery of trachea. Six-month follow-up showed that the patients resumed their normal life. CONCLUSION: The artificial trachea completely healed with the native trachea and became a part of the human trachea. The inner side of artificial trachea is coated with intact native skin tissue with ample blood supply, totally alive without rejection. Therefore, the pedicled artificial tracheal prosthesis is an real artificial trachea. PMID- 12887781 TI - [Diagnosis and treatment of cystic acoustic neuroma]. AB - OBJECTIVE: To study the clinical features and treatment of cystic acoustic neuroma. METHODS: Twenty-two patients with cystic acoustic neuromas were diagnosed by CT and MRI preoperatively, and the tumors were resected by retrosigmoid approach, and verified by pathological studies. RESULTS: Of the 22 patients, 18 had tumors totally resected (postoperative house Brackman facial nerve grading: grade II in 4 patients, III in 7, IV in 3, V in 2, VI in 2) and 4 had tumors subtotally resected. CONCLUSION: Because of its specific clinical features and poor operative results, cystic acoustic neuroma should be regarded as a specific subtype. PMID- 12887782 TI - [Parapelvic cyst of kidney]. AB - OBJECTIVE: To study the clinical features, treatment and diagnosis of parapelvic cyst. METHODS: Twenty-three patients of parapelvic cyst of the kidney were reviewed retrospectively. Fourteen cases (61%) complained of lumbar pain or discomfort, and 4 patients (17%) accompany hematuria and hypertension. RESULTS: In 15 patients receiving surgery, 2 were treated by nephrectomy, one by radical nephrectomy for misdiagnosis. Postoperative diagnosis confirmed a cyst. Eight patients were treated conservatively for cyst being small and without clinical symptoms. Nineteen cases were followed up for 0.5 - 12.0 years. CONCLUSIONS: Ultrasonography and CT scan are the main diagnostic methods. Enhanced CT is extremely helpful in differential diagnosis of hydronephrosis. Surgical management is suitable for big cysts, lumbar pain, hematuria, hypertension and other complications. PMID- 12887783 TI - [Hemodynamometry of erectile dysfunction]. AB - OBJECTIVE: To study the clinical causes of the erectile dysfunction (ED). METHODS: One hundred and thirty cases of ED were examined by hemonamometry and cavernosography with vasodilating agent. The data about penile brachial index, intracavernous pressure, maintenance flow rate, and pressure loss change were obtained and the status of the penile veins was detected. RESULTS: Among 130 patients with ED, 39 had venous leakage including penile arterial insufficiency simultaneous venous leakage in 15 patients. Various leakage sites were observed by using cavernosography. Twenty-eight patients showed deep dorsal veins only and the remaining crural veins. CONCLUSION: Hemodynamometry is effective to diagnose the cause of ED. PMID- 12887784 TI - [Experiment on fibroblast-PGA complexes cultured in rotary cell culture system]. AB - OBJECTIVE: To investigate the effects of RCCS on cell seeding onto 3-D scaffold and cell-scaffold composite culture in vitro. METHODS: Rabbit skin fibroblasts of passage 2 were seeded at 2 x 10(6) cell per cm(3) onto/into polyglycolic acid (PGA) foams by static seeding (dropping a cell suspension onto foams) or dynamic seeding (rotating PGA foams and a cell suspension in RCCS). Attachment of cells in foams was observed by cell-counting after trypsin digestion. The effects of culture condition were next studied by culturing cell-PGA complexes in RCCS versus static culture condition. Distribution and proliferation of cells in foams were investigated with MTT, stereomicroscope and scan electron microscope. RESULTS: Numbers of cells adhering to polymers increased gradually during an initial period of 24 hours. Eight, 12 and 24 hours after seeding, the rates of adhering cells were significantly higher in the dynamic seeding group than in the static seeding group (46.70% + 2.16% vs. 31.50% +/- 3.54%; 56.36% +/- 3.18% vs. 34.28% +/- 3.16%; 66.32% +/- 4.60% vs. 37.38% +/- 4.66%; P < 0.01). The dynamic culture method as compared to the static method resulted in new tissues with a higher cellularity and more uniform cell distribution during a 3 period of weeks. CONCLUSIONS: RCCS has advantages of promoting cell attachment, uniform migration and proliferation in polymer scaffolds and can be used for construction of 3-D cell-polymer tissues in vitro. PMID- 12887785 TI - [Clinical application and animal experiment of thoracic tracheal reconstruction by using pulmonary tissue flap]. AB - OBJECTIVE: To study the usability of pulmonary tissue flap in the reconstruction of the thoracic trachea. METHODS: Over half perimeter anterior and posterior wall and 6 to 8 tracheal cartilagious rings of dog trachea were resected. The nickel titanium alloy mesh stent was placed inside the lumen for repair of the defect of the tracheal wall by nearby pulmonary tissue flap with the vascularized segment. The dogs were killed from 2 months to 12 months after operation. Specimens were taken and observed under light and electron microscope. Clinically, 4 patients were treated by this way operation [right common tracheal scarred stenosis and atresia (1), mixed cancerization in the lower part of the common trachea (1) and left common tracheal carcinoid (2)]. RESULTS: No stenosis and granulation tissue were observed in the prosthetic lumen, in which there was comparative continuous stratified ciliated columnar epithelium. All patients recovered respiratory function. CONCLUSION: Pulmonary tissue flap is a promising prosthesis of thoracic tracheal reconstruction. PMID- 12887786 TI - [Experimental study of canine tracheal allotransplantation]. AB - OBJECTIVE: To detect the factors relevant to stenosis of tracheal graft and to find feasible methods to solve this problem. METHODS: Sixteen mongrel dogs were divided into groups A and B randomly and equally. Five-ring-length tracheal segments were allotransplanted. All grafts and anastomotic sites were covered with omental pedicles. In group A, no immunosuppressant was given and in group B, the recipients were treated with cyclosporine. The animals were sacrificed 4 weeks after operation, and their postmortem specimens were examined grossly and histologically. All allografts were assessed by percent patency. Epithelial regeneration and morphology of the cartilage were semiquantitatively evaluated. RESULTS: Structural integrity of the allografts were maintained better in group B than in group A. Tracheal stenosis was found to be more serious in group A. The scores of epithelial regeneration and cartilage morphology were higher in group B than in group A, and in each group positive correlation was found between the percent patency and the score of epithelial regeneration or cartilage morphology. CONCLUSIONS: Immunosuppressive drugs are necessary to maintain the structure of allografts. Tracheal stenosis is correlated closely with epithelial regeneration and morphological maintenance of the cartilage. PMID- 12887787 TI - [Development of oligoneucleotide arrays for HLA-DR53-associated group genotyping]. AB - OBJECTIVE: To develop a method performed on an oligonucleotide array for HLA-DR53 group genotyping. METHODS: According to the specific allelic frequency and sequence of HLA-DRB loci in Chinese Han population, HLA-DR53 group typing probes which were immobilized on a glass supports were synthesized. A pair of group special primers labeled by the Cy5-dCTP were designed, and the primers were used in the PCR, thus the PCR products were labeled with Cy5. The labeled PCR products were hybridized with array. The signals were scanned by scanner and analyzed by image software. The typing results were confirmed by standard DNA and PCR-SSO. One hundred and eleven samples were typed by this array. RESULTS: There were 72 HLA-DR53 group loci typed by oligonucleotide array. Among them, 34 loci were DR9, 25 were DR4, and 13 were DR7. No false positive or false negative typing results were observed. The specificity and reproducibility were 100% and the overall time of genotyping was 5 hours. CONCLUSION: The oligonucleotide array technique is a precise, rapid molecular method for HLA-DR53 genotyping, suited for clinical practice. PMID- 12887788 TI - [Epidemiological investigation of perinatals affected by transverse facial cleft in China]. AB - OBJECTIVE: To study the epidemiological features of transverse cleft and its multiple congenital anomaly (MCA) patterns in China. METHODS: From 1987 through 1992, hospital-based cluster sampling method was adopted for collecting data. During that period all live or stillbirths with 28 weeks of gestation or more were assessed within 7 days after delivery. RESULTS: 94 cases of transverse facial cleft were identified among 4,489,692 births, so the prevalence rate of transverse facial cleft at birth was 0.21/10(4). The prevalence rates in urban areas and in rural areas were 0.20/10(4) and 0.23/10(4), respectively. And the rates in male and female births were both 0.21/10(4). 69 cases occurred with other malformations, among which the anomalies of ear were the most frequent association (53.6%). The perinatal fatality rate was 46.9%, a significant difference of fatality rate was found between isolated forms (16.0%) and associated forms of transverse facial cleft (58.0%). CONCLUSIONS: The prevalence rate of transverse facial cleft in China is more than the estimated rate. No significant differences are found between urban and rural areas, and between male and female births. Most of transverse facial clefts are associated forms, often as one feature of other syndromes. PMID- 12887789 TI - [The evaluation of the Chinese intelligibility of patients with velopharyngeal incompetence]. AB - OBJECTIVE: To study the Chinese intelligibility of patients with velopharyngeal incompetence (VPI). METHODS: 100 patients with VPI and 32 normal students were selected for this study. Of 10 patients, 15 with cleft palate, 21 with congenital velopharyngeal incompetence, 56 with post-palatoplasty VPI and 8 with post pharyngoplasty VPI. Chinese intelligibility was measured by three speech pathologists and relation between degree of abnormal articulation and Chinese intelligibility were studied. RESULTS: Chinese intelligibility of the controls was 99.0% and the patients with VPI were 35.2%. Of the patients, the cleft palate was 19.9%, the congenital velopharyngeal incompetence was 32.8%, the post palatoplasty VPI was 40.3% and the post-pharyngoplasty VPI was 35.2%. The data showed obvious difference among the controls and sub-types of VPI (P < 0.01). CONCLUSIONS: (1) Chinese intelligibility of VPI accompanied with hypernasality is worse. (2) Of misarticulation of VPI, the cleft palate is the worst, in turn the others are the congenital velopharyngeal incompetence, the post-pharyngoplasty VPI and the post-palatoplasty VPI. PMID- 12887790 TI - [The influence of the osteogenesis in cleft palate after palatal surgery to the transverse growth of dental occlusion]. AB - OBJECTIVE: To compare the transverse growth of the dental cast following palatal surgery with or without osteogenesis in palatal gap. METHODS: 31 patients at 13 - 28 years old with repaired palate were selected to take the dental cast, the width of the dental arch, alveolar base and the matching degree between arches were measured and the average was calculated. After the length of the bone bridge in palate was measured through CT scan, the relationship between the osteogenesis in palatal gap and the transfers growth of dental cast was studied. RESULTS: The critical length of the bone regeneration in palatal gap to affect the dental growth is 4 mm, mean while the sufficient length is 8 mm. The group of sufficient bone bridge has a better development in both the width of dental alveolar base and that of dental arch and the matching degree than the group of unsurficience. And this intendance was more obvious in premolar region than in molar region. CONCLUSIONS: The osteogenesis in palatal gap following palatal surgery can strongly support the transfers growth in the width of dental arch, the width of alveolar base and the matching degree between upper and lower dental arch. PMID- 12887791 TI - [A preliminary study of the two models treated by presurgical orthopedics compared with early soft palate adhesion method]. AB - OBJECTIVE: To compare results of the growth and development of the upper dental arch and the velopharyngeal closure of the cleft patients treated by two methods. METHODS: The dental cast of patient and X-ray films were measured and the statistical medical records were analyzed. RESULTS: The transverse distance of upper dental arch was found to be wider in group A than in group B. The anterior posterior distance of the dental arch in bilateral cleft group was shorter in group A than in group B. The difference of the two groups were gradually lessened as age increases. Bony bridge in alveolar gap was 63% and 83.3% in unilateral and bilateral cleft group respectively. 15% of cases in group A and 35.2% in group B needed pharyngeal flap. CONCLUSIONS: The stable upper dental arch in group A can opposes the pressure from the lip muscles, this maintains the width of the arch. But A-P distance of upper dental arch in BCLP in group A should be followed up after the age of 9 years. Pharyngeal flap is needed less in group A than in group B. PMID- 12887792 TI - [CT assist in surgical design for the posterior pharyngeal flap]. AB - OBJECTIVE: To establish a method for assist design of the posterior pharyngeal flap (PPF). METHODS: The width, length of velopharyngeal space in repose and phonation of "i" in 26 cases with VPI were measured with CT. The width and length of PPF needed, the location of the pedicle and the contact position with the soft palate were analyzed. RESULTS: The width of velopharyngeal space in repose ranged from 15.10 to 37.25 mm, with the average of 27.01 mm, the length was 12.54 to 30.39 mm, with the average of 22.10 mm. The width in phonation of "i" was 9.11 to 34.06 mm, with the average of 17.69 mm, the position of the pedicle wasall located at the level of the first cervical vertebra or above and the contact area of the flap with the soft palate was in the middle posterior 1/3 mostly. CONCLUSION: As a method to quantitatively evaluate velopharyngeal function, CT can assist in design for the PPF. PMID- 12887793 TI - [Comparative morphometrical study on development of palatal shelves in cleft and non-cleft palate mice]. AB - OBJECTIVE: To quantitatively compare the relationship between the congenital cleft palate and development of the palatal shelf. METHODS: Fifty two pairs palatal shelves were macroscopic measured, and 60 series coronal sections were microscopically measured, which were precisely orientated in the coronal plane and serially sectioned at 7 micro m thickness. With the aid of computer imaging analysis system the widths and areas of the palatal shelves in vertical and coronal direction, the maximal areas of the palatal bone and palatal process and alveolar process were measured and compared quantitatively between the cleft group and non-cleft group. RESULTS: The widths and areas of palatal shelves in cleft foetuses showed significant reduction macroscopically and microscopically as well as the maximal areas of the palatal bone, in addition, both of two processes of the maxilla showed significant developmental deficiency. CONCLUSIONS: The palatal shelves show significant developmental hypoplasia in three dimension directions, which have significant correlation between palatal cleft and trisomic condition. PMID- 12887794 TI - [Study on etiology of retinoic acid-induced cleft palate in mouse]. AB - OBJECTIVE: To investigate the morphologic changes of embryonic palatal development exposed to retinoic acid (RA) in mouse, and to detect the significance of the expression of TGFbeta1, TGFbeta3, EGF and BCL2. METHODS: The stage of palatal development was examined by light microscopy. S-P immunohistochemistry and in-situ hybridization was used to detect spatio-temporal patterns of expression of TGFbeta1, TGFbeta3, EGF and BCL2 in embryonic palate. RESULTS: The fetus exposed to RA resulted in formation of small palatal shelves without contact and fusion of each other to form and intact palate. RA can regulate the embryonic palatal expression of genes involved in RA-induced cleft palate. CONCLUSIONS: RA can inhibit the proliferation of MEPM cell to form small palatal shelves and induce abnormal differentiation of MEE cell causing the bi palatal shelves no contact and fuse with each other, then induce the formation of cleft palate. RA can regulate the spatio-temporal patterns of expression of TGFbeta1, TGFbeta3 and EGF in embryonic palatal processes and the change of special expression of these genes in embryonic palatal processes are involved in RA-induced cleft palate. PMID- 12887795 TI - [Study on the expression level of Annexin I, cPLA(2) and PCNA in cleft-palate mice induced by dexamethasone]. AB - OBJECTIVE: To test the changing expression level of Annexin I, cPLA(2) and PCNA in the palatine process of cleft-palate mice A/J and C57B/6j induced by dexamethasone. To discuss the developing mechanism of cleft-palate and the corresponding preventive methods. METHOD: s Pregnant mice A/J and C57BL/6j were randomly divided into normal group as blank control, group with deformity induced by dexamethasone, group given VitB(12) as antagonist to deforming factor, group given only VitB(12). The relative quantity of Annexin I, Cpla2 and PCNA were detected by immunoblotting. RESULTS: Among mice A/J, with the development of palatine process, Annexin I's expression level was increased in the normal group, and other groups showed the similar changes. Annexin I's expression level was significantly higher in group DEX and group DEX + VitB(12) than in normal group and VitB(12) group of, while there was no significant difference between normal group and VitB(12) group. But the changes of cPLA(2) and PCNA expression level was in an opposite direction, with development it decreased in the normal mice's palatine process. In mice C57B/6j there was no significant difference between normal group and group DEX on the measured quantity of Annexin I, cPLA(2) and CONCLUSIONS: Annexin I and cPLA(2) introduce glucocorticoid to induce cleft palate. VitB(12) can not inhibit DEX's enhancing effect on the expression level of Annexin I, but it can antagonize DEX's inhibiting effect on expression level of cPLA(2), which is probably one of the mechanisms why VitB(12) antagonize glucocorticoid's deforming effect. PMID- 12887796 TI - [Clinical evaluation and laboratory study of ultrasonic irrigation of the root canal]. AB - OBJECTIVE: To assess the efficacy of ultrasonic irrigation of the root canal in vivo and in vitro. METHODS: Sixty anterior teeth or premolars with the diagnosis of periapical periodontitis were divided into two groups (syringe irrigation group, group S and ultrasonic irrigation group, group U) and were evaluated clinically. Sixty extracted teeth with single straight canals were selected. Forty of them were divided into two groups and were evaluated histologically. Another twenty teeth were divided into two groups and were evaluated by SEM. RESULTS: Clinically, the number of lateral canals obturated in group U was more than that in group S (P < 0.01). The incidence of pain during irrigation were 13.3% in group S, and 3.3% in group U (P > 0.05). Histologically, the amount of organic debris of the root canals in group U were significantly less than that in group S (P < 0.01). By SEM study, the number of visible open dentinal tubules in group U were significantly greater than that in group S (P < 0.05). CONCLUSION: Ultrasonic irrigation of the root canal will be a useful technique in root canal therapy. PMID- 12887797 TI - [A comparative study of masticatory efficiency and mandibular movement pattern among patients with CD, COD and IOD]. AB - OBJECTIVE: To compare the masticatory efficiency and the character of mandibular movement trace of the patients with complete denture (CD), complete overdenture (COD) and implant-supported overdenture (IOD). METHODS: A total of 42 subjects were rehabilitated with CD, COD and IOD respectively and wore the dentures at least half a year. Each study group was composed of 14 subjects. The masticatory function of patients were investigated by means of masticatory efficiency and mandibular movement trace. The characters of mandibular movement trace were evaluated from four aspects: the regulation of chewing cycle, centralization of end traces, the type of the contact slides and the type of the trace on the frontal plane. RESULTS: The average chewing efficiency of almond and jujube in the IOD patients was higher than that in the CD group (P < 0.01), and the chewing efficiency of almond in COD group was higher than that in the CD group (P < 0.01). In the IOD group, there were more patients with regular chewing cycle (P < 0.05) and centralized end traces (P < 0.01) in mandibular movement than those in the CD group and COD group. CONCLUSIONS: IOD and COD benefit the fully or partially edentulous patients with a relatively good masticatory function. The patients with IOD have more regular mandibular movement than those with CD. PMID- 12887798 TI - [Clinical evaluation of 546 tetracycline-stained teeth treated with Cerinate laminate veneers]. AB - OBJECTIVE: The purpose of this study was to evaluate the clinical result of 546 tetracycline-stained teeth corrected with Cerinate porcelain laminate veneer system for esthetic purpose. METHODS: 546 tetracycline-stained teeth were corrected with Cerinate porcelain veneer system, and bonded with Ultra Bond resin cement. The restorations were recalled after 0.5 year, 1.5 years and 2.5 years, respectively. modified Ryge criteria were used to evaluate the veneers marginal adaptation, interfacial staining, secondary caries, postoperative sensitivity and the patients' satisfaction with the shade of the restorations. RESULTS: this study found that 99% veneers had excellent marginal adaptations; and less than 1% veneers were rebonded after debond in the first half year application; the color of the veneers was stable and no evident staining was found. Almost all patients were satisfied with their restoration color after 1 year's application. CONCLUSION: The research indicated that Cerinate porcelain veneer restoration system is a reliable and ideal choice for the correction of tetracycline-stained teeth. PMID- 12887799 TI - [Effect of particle size distribution of dental poly (methyl methacrylate) beads on the microstructure of denture]. AB - OBJECTIVES: To investigate the effect of particle size distribution (PSD) of dental poly (methyl methacrylate) (PMMA) beads on the microstructure of denture. METHODS: It was compared and analyzed that the porosity of three groups of spherical specimens which were made of PMMA beads with different PSD at proper polymer/monomer ratios. And the final microstructure of each group was investigated using Digital High Definition Microscope. RESULTS: With proper polymer/monomer ratio increasing, the occurrence of porosity tended to decrease. The final microstructure of denture varied with PSD of PMMA beads. CONCLUSION: By means of optimizing PSD of beads, the microstructure of denture could be improved, and the occurrence of porosity could be reduced. PMID- 12887800 TI - [Effect of spirochetes and porphyromonus gingivalis on oral malodor]. AB - OBJECTIVE: To assess the effect of Spirochetes and Porphyromonus gingivalis (Pg) on the production of volatile sulfur compounds (VSCs) and oral malodor, respectively. METHODS: 117 systemically healthy periodontitis patients were divided into two groups according to organoleptic rating (OR) by organoleptic method: 85 subjects with oral malodor (halitosis group, HG) and 32 cases without oral malodor (no-halitosis group, NHG). VSCs measurements were made with halimeter. Subgingival plaques and tongue coating were taken to calculate the proportion of Spirochetes (with 2% Congo Red Smears, S%) and the amounts of Pg (with indirect immunofluorescence method). The examination will be repeated in 46 HG cases after periodontal therapy. RESULTS: S% in PD < 4 mm sites of HG was significantly higher than that in NHG (P < 0.01). S% in PD >or= 4 mm sites and tongue coating, Pg in all three sites were not different between two groups. In HG, OR and VSCs levels were correlated with S% in tongue coating (P < 0.01, P < 0.05) and PD < 4 mm sites (P < 0.05), and not correlated with S% in PD >or= 4 mm sites. OR was related to Pg in PD >or= 4 mm sites (P < 0.05) and there was no relationship between OR and Pg in tongue coating and PD < 4 mm sites. VSCs levels was not correlated with Pg in all three sites. S% in tongue coating and subgingival plaque, Pg in subgingival plaque, OR and VSCs levels were significantly reduced after periodontal therapy (P < 0.01). CONCLUSIONS: The direct relationship of Spirochetes and Pg respectively to bad breath was not found obviously. But Spirochetes is one of the microorganism on oral malodor. Periodontal therapy can improve oral malodor. PMID- 12887801 TI - [The expression of Sonic Hedgehog during late bell stage by Western Blot and in situ hybridization]. AB - OBJECTIVE: To investigate the expression and biological effect of Shh during late bell stage by morphological and semi-quantitative analysis. METHODS: Tooth germs were selected from new born Bal b/c mouse (P1, P2, P3, P5, P7). Semi-quality of Shh was measured by Western Blot and the expression place and strength of Shh were observed by in situ hybridization. RESULTS: Shh was expressed in the ameloblast layer during late bell stage; the expression strength was high in secretive period and decreased with development; the active N-section was detectable before P3. CONCLUSION: Shh expresses specially in the ameloblast layer in late bell stage, and expression quality is related to the function of ameloblasts. PMID- 12887802 TI - [Comparative study of friction and wear behavior of different human enamel in vitro]. AB - OBJECTIVE: To investigate the friction and wear behavior of different human enamels. METHODS: 24 enamel samples selected from aged, young permanent and faded deciduous teeth were classified into 3 groups and slid against artificial porcelain teeth in the presence of artificial saliva on an oscillating friction and wear test rig. The wear volume loss, microhardness and toughness of each group of the enamel specimens were measured, the wear scars were observed with a scanning electron microscope, and the elemental compositions of Ca, P, and Si of the wear scar and wear debris were determined with an energy dispersion spectrometer. RESULTS: The wear volume losses of aged, young permanent and deciduous tooth enamels are (2.40 +/- 1.10) x 10(-12) m(3), (3.50 +/- 1.83) x 10( 12) m(3) and (4.86 +/- 2.49) x 10(-12) m(3). The data of aged tooth enamels are statistically greater than that of deciduous tooth enamels (P < 0.05). There is no significant difference between the wear volume loss of aged and young permanent tooth enamels or between the young permanent and deciduous tooth enamels (P > 0.05). However, the friction and wear behavior of each group of enamel specimens is different from each other. CONCLUSIONS: In the present testing condition, the wear scars of each kind of enamel specimens is characterized by ploughing and cracking. The different wear resistance of the three kinds of enamels is attributed to the different microstructure of the enamel, while the hardness and toughness of the enamels are not correlated with the wear resistance. PMID- 12887803 TI - [The effect of overdose fluoride on the proliferation and apoptosis of rat's incisor ameloblast]. AB - OBJECTIVE: To observe the effect of overdose fluoride on the proliferation of rat's incisor ameloblast. METHODS: 20 Wistar rats were divided randomly into 2 groups: Group I (Control); Group II 50 mg/L F(-) were given. After 8 weeks treatment, the AgNORs stain and TUNEL technique were applied to analyze the effect of fluoride on the proliferation and apoptosis of ameloblasts. RESULTS: The imagination analysis results showed that proliferation of pre-secretion ameloblasts were inhibited in group II as compared with the control group (P < 0.001). There was significant increase of apoptosis with the trend of migration toward secretion stage. CONCLUSION: The mechanism of fluorosis mottled enamel may be the effect of overdose fluoride with inhibits proliferation and induces apoptosis of ameloblasts resulting in dysfunction of secretion or absorption of enamel matrix proteins. PMID- 12887804 TI - [Inhibition of remineralization by EDTA-soluble phosphate protein in dentin]. AB - OBJECTIVE: To investigate the effect of removing EDTA-soluble phosphate protein in dentin on the later remineralization for the purpose of better understanding of mechanism of dentin phosphate proteins on dentin mineralization. METHODS: To remove soluble phosphate protein by EDTA dissolution, then the remineralization rate was monitored by a constant composition crystal growth technique. The results were compared with those from the normal dentin and the dentin partially demineralized by acetic acid. RESULTS: Faster remineralization rates were found with dentin demineralized by EDTA (0.5 and 2 h) compared with normal dentin powder, while a slower rate was found with dentin demineralized by acetic acid. The increase of remineralization rate by removing phosphate protein from dentin was 100% more at 200 min after the start of the reaction. CONCLUSION: EDTA soluble phosphate protein in dentin has a great potential to inhibit remineralization. PMID- 12887805 TI - [Rapid detection of Streptococcus mutans and streptococcus sobrinus in human saliva by nested polymerase chain reaction]. AB - OBJECTIVE: To establish a simple and rapid method to detect Streptococcus mutans and streptococcus sobrinus simultaneously in human saliva. METHODS: Chromosomal DNA from the bacteria was obtained by the extraction method with phenol chloroform. A nested PCR method with two sets of primers specific for portions of the glucosyltransferase genes (gtfB of S. mutans and gtfI of S. sobrinus), was optimized to detect S. mutans and S. sobrinus from standard strains, clinical strains and directly in human saliva. RESULTS: The first process of nested PCR was capable of amplifying DNA fragments specific for these species from chromosomal DNA extracted from 10(5) CFU cells of standard and clinical strains, or from 1 ml clinical saliva samples containing 10(5) CFU cells of either species. a second process of nested PCR, using the first PCR product as a template with new internal primers to detect 10(3) CFU of either streptococcal species in 1ml saliva samples. CONCLUSION: Nested PCR could detect S. mutans and S. sobrinus rapidly and simply in human saliva. This finding would be important to studies of elucidation the role of these two streptococcal species in the etiology of dental caries. PMID- 12887806 TI - [Gene expression and pathological changes of rat submandibular glands after adenovirus-mediated gene transfer]. AB - OBJECTIVE: To investigate luciferase gene expression and pathological changes of submandibular glands (SMG) of rats after adenovirus-mediated gene transfer. METHODS: Adenovirus-mediated luciferase gene (AdCMVLuc, 10(8) pfu in 50 microl) was injected in to SMG of forty wistar rats. The SMGs were harvested for gene expression measurement and pathological study after 3 days, 1,2,4 and 8 weeks. RESULTS: Peak expression was observed in three days following administration of the vector however, gene expression in submandibular glands decreased rapidly. the pathological changes induced by retrograde injection of AdCMVLuc included: after 3 days to one week, compression of acini, dilation of terminal ducts; after two weeks, slight atrophy of a part of acini, increase of iteracinar distance and focal lymphocyte infiltration in lobules and interlobular ducts; after 4 weeks, recovery evidence was found in acini; after 8 weeks, normal acini and ducts were found. The ultrastructural changes included: 3 days, much more rough endoplasmic reticulum was found both in acini and duct epithelial cell; a lot of mucus drops were found in acini; after 1 week, microvillus decreased in duct epithelial cells, mitochondria increased significantly in acini; intercellular space was enlarged. CONCLUSIONS: Adenovirus-mediated gene transfer can produce biological proteins and induce marked inflammatory changes in rat SMG. The ultrastructural changes suggest high protein synthesis activity in the acinar cells after gene transfer. PMID- 12887807 TI - [The inhibitory effect of glass ionomer cement for orthodontic bonding on bacteria]. AB - OBJECTIVE: To prove the inhibitory effect of glass ionomer cement for orthodontic bonding on bacteria. METHODS: Twenty-two persons were chosen randomly. In vivo, with the split-mouth technique, brackets were bonded with glass ionomer cement or composite resin respectively. Plaque samples were taken from the area of the adhesive one month after the placement of brackets and incubated for microbiologic analysis. RESULTS: There was no significant difference in the amount of total anaerobic bacteria with the two materials. There were also no obvious reduction of the numbers of S. mutans (the composite: 2.4 x 10(10) CFU/L, the glass ionomer cement: 2.8 x 10(10) CFU/L, P = 0.673) and its proportion (percentage of total CFU) with the use of glass ionomer cement compared with the composite. CONCLUSIONS: There was no long term inhibitory effect on bacteria adjacent to glass ionomer cement for orthodontic bonding. PMID- 12887808 TI - [A comparative study of the bracket bond failure between adults and adolescents receiving fixed orthodontic therapy]. AB - OBJECTIVE: To evaluate the bracket bond failure and its causes between adult and adolescent patients during fixed orthodontic therapy. METHODS: Bracket bond failure data of 30 adults and 30 adolescents, receiving fixed orthodontic therapy, have been collected within the first 12 visits, respectively. The compliance has been analyzed with survival analyse between the two groups. RESULT: The general bracket bond failure rate in the adult group is lower than that of the adolescent group and the difference is significant (p < 0.05). In the adolescent group, the failure rate for mandibular anterior teeth is highest and different from that of the adult group (p < 0.05). The failure rate resulted from biting hard food is ranked No. 1. CONCLUSIONS: The compliance of the adults receiving fixed orthodontic therapy is better than that of the adolescents. The investigation of bracket bond failure causes is beneficial in helping orthodontists improve orthodontic practice and raise clinical efficiency. The survival analysis is effective in evaluating the bond failure. PMID- 12887809 TI - [The relationship of wire offset and lingual component force on first molar]. AB - OBJECTIVE: To measure the strength of lingual component force on first molar while retraction canine by 150 g tractive force with different sizes of arch wire and with different widths of molar offset on the wire. METHODS: Wires with different sizes and different materials [0.40 mm and 0.45 mm stainless round wires, 0.40 mm and 0.45 mm Australia wires, 0.41 mm x 0.56 mm (0.016 x 0.022 inch) and 0.46 mm x 0.63 mm (0.017 x 0.025 inch) rectangular stainless wires] were ligated in ideal arch Typodont when the canine was retracted with 150 g force in the direction from the molar buccal hook to the central point of canine bracket's distal wing under different widths of molar offset (1 mm, 2mm, 3mm). the lingual force applied on the first molar was measured by a self-made dynamometer. RESULTS: 1. There is lingual component force applied to the first molar while retracting canine with 150 g tractive force. 2. the bigger the cross section area of the wire and the stronger the rigidity of the wire, the smaller the lingual component force turns. The ability of the wire in deducting the lingual component force is in direct ratio with the cross section area and the rigidity of the wire. 3. While the width of wire offset increases, the lingual force produced by the tractive force applied to the first molar decreases lineally. CONCLUSION: There is lingual force produced on the anchorage molar when retracting canine. In clinic for avoiding the lingual movement of the first molar, increasing the size of wire or increasing the width of wire offset is suggested. PMID- 12887810 TI - [Citation analysis of part of features in the Chinese Journal of Internal Medicine and other 3 foreign journals]. AB - OBJECTIVE: To analysis the discrepancy of citations in different features of the Chinese Journal of Internal Medicine (Chin J Intern Med) and other 3 foreign journals (BMJ, JAMA, Ann Intern Med). METHODS: Citations of articles published from 1998 to 2002 in major features of the four journals were calculated. RESULTS: In the 5 years, the average number of citation original articles in Chin J Intern Med was 6.55, in BMJ 15.78, in JAMA 34.73 and Ann Intern Med 30.37; that of commentary articles was 3.82, 9.26, 18.38 and 13.79, respectively. In the recent five years, 86% citations were devoted to English language articles, 14% Chinese language articles, less than 1% Japanese articles, few other language articles. Price index analysis indicated the value for BMJ, JAMA, Ann Intern Med and Chin J Intern Med was 0.58, 0.57, 0.42 and 0.50, respectively. CONCLUSIONS: The discrepancy in average number of citation articles in different features of the four journals is not significantly different, even in time of their citation. The citation of articles is not varied in languages in Chin J Intern Med. PMID- 12887811 TI - [Clinical characteristics of non-steroidal anti-inflammatory drugs associated gastroduodenal ulcer bleeding]. AB - OBJECTIVE: To study the clinical characteristics of non-steroidal anti inflammatory drugs (NSAIDs) associated gastroduodenal ulcer bleeding. METHODS: Patients who were hospitalized in our ward because of gastroduodenal bleeding from Jan, 1997 to Dec, 2001 were divided into two groups according to consumption of NSAIDs or not in the week previous to the onset of bleeding. RESULTS: 446 patients were investigated with 86 in NSAIDs group and 360 in non-NSAIDs group. There was no significant difference in sex, way of bleeding, history of peptic ulcer, bleeding site in stomach or duodenum, presence of erosion and the need of endoscopic injection therapy or not between the two groups. However, the patients in NSAIDs group were older than those in non-NSAIDs group. Hemoglobin level was lower in NSAIDs group (P = 0.004). There was more gastric ulcer and complex ulcer in the NSAIDs group than in non-NSAIDs group (P < 0.001). NSAIDs users had more ulcers (P < 0.001). Helicobacter pylori (Hp) was present in 72.5% of the non NSAIDs group and 53.4% of the NSAIDs group (P = 0.001). In an another study, we found that age of the patients or Hp state didn't affect the clinical characteristics of NSAIDs ulcer bleeding. CONCLUSIONS: The clinical characteristics of NSAIDs associated gastroduodenal bleeding should be better understood so as to decrease the occurrence of NSAIDs ulcer and its complication. PMID- 12887812 TI - [Long-term efficacy of lactulose in patients with subclinical hepatic encephalopathy]. AB - OBJECTIVE: To observe the long-term clinical efficacy of lactulose in patients with subclinical hepatic encephalopathy (SHE) and the natural history of this disease. METHODS: Patients with SHE diagnosed with psychometric tests (number connection test and digit symbol test) were randomly allocated to control group (Cg), short-term (Stg) and long-term treatment group (Ltg), respectively. Lactulose (15 - 90 ml/day) was orally given to 21 patients in Stg for 8 weeks, and 22 in Ltg for 24 weeks.The drug was not given to 21 patients in Cg. Psychometric test, somatosensory evoked potentials (SEP) and venous blood ammonia concentration (BA) were investigated before and after treatment at 8 week intervals for 6 months of follow-up. RESULTS: In Ltg, psychometric tests were significantly improved, BA was lowered and N(50) peak latency in SEP was not prolonged as compared with those in Cg, in which psychometric tests and N(50) peak latency gradually worsened during the follow-up. After 8 weeks of treatment, psychometric tests, BA and N(50) peak latency were significantly improved in Stg, but at week 24 they were deteriorated after cessation of treatment. The development of clinical hepatic encephalopathy was more frequent in Cg (40%) and Stg (30%) than in Ltg (5%). One patient each was not followed up after week 24 in Cg and Stg, respectively and 2 patients in Ltg stopped lactulose because of severe diarrhea. CONCLUSIONS: In patients with SHE, long-term treatment with lactulose could lower BA, improve psychometric tests, prevent SEP from getting worse, and eventually cut down the occurrence rate of clinical hepatic encephalopathy. PMID- 12887813 TI - [Default of KIAA0353 gene in varicose great saphenous vein accompanying primary deep vein valve insufficiency]. AB - OBJECTIVE: To screen and identify the genes related to the occurrence and development of varicose great saphenous vein in the patients with primary deep vein valve insufficiency (PDVI). METHODS: mRNA fluorescent differential display (FDD) technique was used to compare the different cDNA fragments originated from differentially expressed mRNAs from the venous tissues of 10 patients with varicose great saphenous vein complicated with PDVI. Ten specimens of normal venous tissue from 10 patients dying from other diseases were used as controls. The differently expressed cDNA fragments were then re-amplified and labeled with DIG to prepare probes for later Northern blotting. Positive fragments confirmed by Northern blotting were cloned into pGEM-T easy vector and sequenced using Sanger's method. Then the sequences were compared with the data in GeneBank by BLASTN software to search for their genetic origin. RESULTS: Altogether 37 differentially expressed cDNA fragments were discovered from the 2 groups, among which 30 were confirmed by Northern blotting. There was a notable 540 bp-long cDNA fragment, which was only presented in the control group, sharing 99% homology with part of the mRNA sequence of human KIAA0353 gene. CONCLUSION: The varicose great saphenous vein of PDVI patients is a process with the involvement of multiple genes and the default of KIAA0353 gene may play a role in the occurrence and development of varicose great saphenous vein in PDVI patients. PMID- 12887814 TI - [Relationship of atherogenesis and proteoglycans in wall of vein graft, an experimental study]. AB - OBJECTIVE: To investigate the relationship between the concentrations of proteoglycans (PGs) and progress of atherogenesis in grafted vein. METHODS: A section of common carotid artery with a length of 0.5 cm was cut off and then a section of vein was implanted to the damage among 72 male New Zealand rabbits. Then the rabbits were randomly divided into two groups of 36 rabbits to be fed with high fat diet and ordinary diet respectively. Every six rabbits were killed one week, 2 weeks, 4 weeks, 8 weeks, 12 weeks, and 20 weeks after the operation. The implanted veins were taken to be observed by electron microscopy. The PGs in the implanted veins were extracted and divided into heparan sulfate proteoglycan (HSPG), chondroitin sulfate proteoglycan (CSPG), and dermatan sulfate proteoglycan (DSPG) by anion-exchange chromatography 8, 12, and 20 weeks postoperatively. The concentrations of PGs were measured. Blood was extracted from the heart to examine the level of LDL-cholesterol. The normal jugular veins on the opposite side were used as controls. RESULTS: The level of LDL-cholesterol in the high fat diet group was 26.5 +/- 7.49 mmol/L, significantly higher than that in the normal diet group (0.71 +/- 0.65 mmol/L, P < 0.05). In the normal diet group, the content of CSPG-DSPG was 1,077 +/- 116 micro g/g, significantly higher than that of the normal vein (809 +/- 75 micro g/g, P < 0.05), and not significantly different from that of the normal diet group (1,077 +/- 116 micro g/g) 8 weeks postoperatively, and became lower 12 and 20 weeks postoperatively (773 +/- 49 and 819 +/- 45 micro g/g respectively), both similar to that of normal vein (both P > 0.05). Foam cells were found 20 weeks postoperatively. In the high fat diet group, the content of CSPG-DSPG was 1,089 +/- 94 micro g/g 8 weeks postoperatively, significantly higher than that of normal vein (P < 0.05), and the high level lasted till the 20 th postoperative week (1,068 +/- 100 micro g/g, P < 0.05). Foam cells were detected as early since the 4 th postoperative week, unorganized areas were found 20 weeks after the operation. The HSPG content was not significantly different among the high fat diet group, normal diet group, and the normal vein (with a range of 213 +/- 34 - 278 +/- 33 micro g/g), The proportion of HSPG in total PGs 8, 12, and 20 weeks postoperatively in the high fat diet group were 19.6%, 18.6%, and 16.6% respectively, lower than those in the normal diet group (18.8%, 21.9%, 21.9%) and much lower than that in the normal vein (25.55%). CONCLUSION: CSPG-DSPG has auxo-action on atherogenesis in graft, but HSPG protects the graft against atherogenesis to an extent. PMID- 12887815 TI - [The relationship between hypoxia inducible factor-1 alpha expression and novoendotheliasis after autogenous vein grafting]. AB - OBJECTIVE: To elucidate the relationship between the level of hypoxia inducible factor-1alpha (HIF-1alpha) expression and novoendotheliasis after autogenous vein grafting. METHODS: 40 Wistar rats underwent right internal jugular vein to common jugular artery interposition grafting. Twenty rats were killed 7 and 14 days after the grafting respectively and the grafted veins were harvested respectively. Twenty rats not undergoing grafting were used as controls. The expressions of HIF-1alpha mRNA and protein and vascular endothelial growth factor (VEGF) mRNA and protein were examined by using reverse transcription polymerase chain reaction, Western blotting, and immunohistochemistry. Electron microscope was used to observe the process of vascular novoendotheliasis. RESULTS: The percentage of HIF-1alpha positive cells 7 days after grafting was 20.5 +/- 6.2%, significantly higher than that 14 days after (12.3 +/- 3.7, P < 0.01) and that of the controls (0.5 +/- 0.1%, P < 0.01). The percentage of HIF-1alpha positive cells 14 days after grafting was significantly higher than that of the controls too (P < 0.01). The expression of HIF-1alphamRNA 7 days after grafting was 53.2 +/- 9.3, significantly higher than that 14 days after grafting (43.2 +/- 6.3, P < 0.01) and that in the control (29.4 +/- 5.6, P < 0.01). The expression of HIF 1alpha7 mRNA 14 days after grafting was significantly higher than that of the controls too (P < 0.01). The expression of HIF-1alpha protein 7 days after grafting was 5.6 +/- 0.4, significantly higher than that 14 days after grafting (3.5 +/- 0.2, P < 0.01) and that in the control (0.8 +/- 0.1, P < 0.01). The expression of VEGF protein 7 days after grafting was 2.2 +/- 0.1, significantly higher than that of the control (0.5 +/- 0.1, P < 0.01). Novoendotheliasis was seen 7 days after grafting and decreased 14 days after. The expression of HIF 1alpha was positively correlated with the expression of VEGF (r = 0.903, P < 0.01). CONCLUSION: HIF-1 alpha is closely related to novoendotheliasis. The deficiency of HIF-1 alpha expression plays an important role in intima hyperplasia. PMID- 12887816 TI - [Chlamydia-like and coronavirus-like agents found in dead cases of atypical pneumonia by electron microscopy]. AB - OBJECTIVE: To explore the causative agents of the atypical pneumonia (also SARS) occurred recently in some regions of our country. METHOD: Organ samples of 7 dead cases of SARS were collected from Guangdong, Shanxi, Sichuan Provinces and Beijing for electron microscopic examination. 293 cell line was inoculated with the materials derived from the lungs to isolate causative agent(s). The agents in the organs and cell cultures were revealed by immunoassay. RESULTS: Both Chlamydia-like and coronavirus-like particles were found in EM. Inclusion bodies containing elementary bodies, reticulate antibodies and intermediate bodies of Chlamydia-like agent were visualized in multiple organs from the 7 dead cases, including lungs (7 cases), spleens (2 cases), livers (2 cases), kidneys (3 cases) and lymph nodes (1 cases), by ultrathin section electron microscopy (EM). In some few sections, coronavirus-like particles were concurrently seen. A coronavirus RNA- polymerase segment (440 bp) was amplified from the lung tissues of two cases of the SARS. After inoculated with materials from the lung samples, the similar Chlamydia-like particles were also found in the inoculated 293 cells. Since the Chlamydia-like agents visualized in both organs and cell cultures could not react with the genus specific antibodies against Chlamydia and monoclonal antibodies against C. pneumoniae and C. psittaci, the results might well be suggestive of a novel Chlamydia-like agent. CONCLUSION: Since the novel Chlamydia-like agent was found co-existing with a coronavirus-like agent in the dead cases of SARS, it looks most likely that both the agents play some roles in the disease. At the present time, however, one can hardly determining how did these agents interact each other synergetically, or one follows another, need further study. PMID- 12887817 TI - [Induction of angiogenesis in ischemic myocardium by adenovirus mediated angiopoietin-1 gene transfer, an experimental study]. AB - OBJECTIVE: To investigate the effect on angiogenesis in ischemic myocardium of adenovirus mediated angiopoietin-1 gene transfer. METHODS: Ang-1 cDNA was obtained from human spleen by RT-PCR and then cotransfected into 293 host cells together with Adv5-CAG, E1 and E3 substituted adenovirus genome, thus constructing recombinant adenovirus Adv5-CAG/Ang-1. Recombinant adenovirus Adv5 CAG/LacZ containing LacZ gene was constructed too. Thirty six male New Zealand white rabbits were randomly divided into 3 group of 12 rabbits: DMEM group, Ang-1 group, and LacZ group and underwent high-positioned double-ligation of the left anterior descending coronary artery and then myocardial injection of DMEM, Adv5 CAG/Ang-1, or Adv5-CAG/LacZ respectively. Fourteen days after the operation 2 rabbits in each group were killed, the myocardial tissues where injected was given were taken to detect the expression of Ang-1 by RT-PCR. Coronary angiography was conducted 28 days postoperatively upon 5 rabbits in each group to observe the angiogenesis in the ischemic myocardium. Five rabbits in each group were killed at the 14 th and 28 th postoperative days to observe the capillary density by immunohistochemical staining. RESULTS: The Ang-1 cDNA cloned in the laboratory was 1,515 bp in length containing the signal peptide structure in accordance with the report in literature. Fourteen days after operation, Ang-1 gene was positive in the myocardium of Ang-1 group and negative in the other 2 groups. New vessel formation was obvious at the 28 th postoperative day in the Ang-1 group and not in the other 2 groups. Capillary density increased after operation in all 3 groups, however, more significant in the Ang-1 group, especially 28 days after. CONCLUSION: Adenovirus-mediated angiopoietin-1 gene effectively promotes the neovascularization in ischemic myocardium of rabbits. PMID- 12887818 TI - [Forecast about number of patients with pulmonary tuberculosis 2001 - 2010]. AB - OBJECTIVE: To forecast the number of patients with pulmonary tuberculosis in 2010. METHODS: A mathematical model was established based on the nationwide epidemiological survey on tuberculosis conducted in 2000 so as to forecast the numbers of patients with pulmonary tuberculosis in 2000s. RESULTS: (1) The number of patients with pulmonary tuberculosis would be a little more than that in 2000, with the pulmonary tuberculosis case detection rate rho of 0.26 being adopted. (2) The number of patients with pulmonary tuberculosis would be smaller than that in 2000, with the pulmonary tuberculosis case detection rate rho of 0.30 being adopted. (3) If the current intervention strategy manages to keep the pulmonary tuberculosis case detection rate at the level of 0.35, the decline in number of patients with pulmonary tuberculosis will approach the goal set by the national program that the number of patients with pulmonary tuberculosis be decreased by 50%. CONCLUSION: The goal set by the national program can be achieved only when the pulmonary tuberculosis case detection rate reaches 0.35. PMID- 12887819 TI - [HLA-DRB and -DQB allele contribution to narcolepsy susceptibility in Chinese patients with narcolepsy]. AB - OBJECTIVES: To study the association of narcolepsy with HLA class II alleles in Chinese narcoleptic patients. METHODS: 31 patients with narcolepsy underwent brain computed tomography (CT) scan and magnetic resonance imaging (MRI) testing. All patients received a MSLT test following a routine night's sleep, and serological HLA typing for HLA DR(2). 21 patients received PCR-SSP HLA DR and DQ typing. RESULTS: All patients had sleepiness and cataplexy. There was no evidence for other functional or structural diseases. Sleep paralysis was elicited in 45%; hypnagogic hallucinations, in 61%. Mean sleep latency on MSLT was 2.1 min +/- 1.3 min; sleep-onset rapid eye movement (SOREM) occurred during 2/5 naps in 30 of 31 patients. The average number and latency of SOREM episodes were 4.2 +/- 1.0 episodes and 4.0 min +/- 1.8 min, respectively. All patients but one were HLA DR(2) positive and 86% were HLADRB(1) * 1501-HLADQB(1)*0602 positive. CONCLUSIONS: HLA DR(2) and HLADQw6 are markers for narcolepsy-cataplexy in Chinese. PMID- 12887820 TI - [Effects of iodophor on the thyroid glands of female medical workers]. AB - OBJECTIVE: To investigate the long-term effects of iodophor on thyroid function and autoimmunity in females. METHODS: 119 medical personnel who have been using iodophor as preoperative skin disinfectant more for than 2 years and 123 medical personnel who have not exposed to iodophor were studied. The urinary iodine, thyroid-stimulating hormone (TSH), free triiodothyronine (FT(3)), free thyroxine (FT(4)), thyroid peroxydase antibody (TPO-Ab) and TG-Ab in serum were measured and the thyroid were examined by B mode ultrasound apparatus. RESULTS: The median urinary iodine (MUI) was 300.4 micro g/L in the exposed group, not significantly higher than that in the non-exposed group (269.1 micro g/L, P > 0.05). The TSH levels of the exposed group and non-exposed group were 1.66 mU/L and 1.62 mU/L (P = 0.84). B mode ultrasound examination showed that the prevalence rate of thyroid disorders was 3.36% in the exposed group, not significantly different from that in the non-exposed group (2.44%). The thyroid autoantibody positive rate was not significantly different between these two groups too. The titer of autoantibody in the exposed group was 29.5 IU/L, significantly higher than that in the non exposed group (22.4 IU/L, P = 0.048). The titer of TG-Ab in the exposed group was 21.85 IU/L, not significantly different from that in the non-exposed group (18.7 IU/L, P = 0.542). The mean titer of TPO-Ab in exposed group was 29.5 IU/L, significantly higher than that in non-exposed group (22.4 IU/L, P = 0.048). The total prevalence rate of all sorts of thyroid disorders in the exposed group was 11.76%, significantly higher than that in non-exposed group (4.07%, P = 0.026). The thyroid disorders discovered included clinical hyperthyroidism and subclinical hypothyroidism. CONCLUSION: Using iodophor as preoperative skin disinfectant may result in an increased incidence of thyroid disorders in female medical personnel. Medical personnel with iodophor exposure history should have their thyroid function and thyroid autoimmune status examined regularly. PMID- 12887821 TI - [Mechanism of translocation between chromosomes 4q and 10q in facioscapulohumeral muscular dystrophy]. AB - OBJECTIVE: To investigate the translocation between chromosomes 4q and 10q and its putative correlation with facioscapulohumeral muscular dystrophy (FSHD). METHODS: Double digestion of the genomic DNA of 50 controls and 45 FSHD cases, 16 cases of sporadic FSHD and 19 cases of familial FSHD from 7 families, with restriction enzymes BglII and BlnI was followed by separation and Southern blotting with the probe p13E-11. By Scion Image program, the density of hybridized fragments was analyzed and the 4q:10q density ratio was then calculated to infer the types of 4q-10q translocation. The frequencies of translocation among the controls, sporadic FSHD cases, and familial cases were compared. RESULTS: In the 95 subjects 4 different translocation types were detected with a translocation rate of 17.89%. Both the rate of 4q-->10q translocation and the rate of 10q-->4q translocation were 8.0% among the healthy controls. Only 4q-->10q translocation was found in sporadic FSHD cases with a frequency of 43.75%, while only 10q-->4q translocation was found in familial FSHD patients with a frequency of 6.89%. The frequency of 4q-->10q translocation in the sporadic FSHD cases was significantly higher than that in the control group (chi(2) = 11.154, P < 0.001), while the frequency of 10q-->4q translocation in the familial FSHD cases was not significantly different from that in the controls (chi(2) = 0.012, P > 0.5). CONCLUSION: Frequent translocations between chromosomes 4q and 10q occur in normal population and FSHD cases, while the 4q- >10q translocation is only related to sporadic FSHD. Excessive loss or conversion (to 10q26) of 4q35 D4Z4 repeats, caused by 4q-10q interactions, may be essential for the mechanism of this disorder. PMID- 12887822 TI - [Clinical application for reference strand mediated conformation analysis system in HLA-A genotyping]. AB - OBJECTIVE: To establish and stabilize a new HLA typing strategy, reference strand mediated conformation analysis (RSCA), and to conform its advantages by using HLA A as target gene. METHODS: 20 standard DNA samples from international RSCA cooperative team were used to establish and stabilize RSCA. 84 DNA samples of related hematopoietic stem cell transplant donor-recipients were extracted from peripheral blood cells. HLA-A loci were typed both by RSCA and PCR-SSP. RESULTS: RSCA results of 20 standard samples were identical to those of international RSCA cooperative team, and the accuracy and replication rate were both 100%. Among the 84 samples, 82/84 (97.6%) cases could be designated definitely, and 33/84 (39%) cases could be typed to allelic level. In addition, 2/84 (2.4%) cases could not be detected by RSCA for their poor PCR results. 20 samples were randomly selected to identify the replication rate of RSCA, and the results demonstrated that the replication rate was 100%. Among the PCR-SSP typing results, there were about 10% samples needed to be typed for 1 - 3 times to confirm their types. CONCLUSION: RSCA has some advantages compared with the PCR-SSP typing method, which are high resolution, high sensitivity, high accuracy, high replication, capability to find new alleles, high throughput, low cost and suitability for unrelated hematopoietic stem cell transplant donor-recipient HLA typing. But there are still some defects in this new strategy, which are time-consuming for only one sample a time and higher DNA quality requirement. In addition, RSCA database needs to be further improved. PMID- 12887823 TI - [Relationship between regression of hypertensive left ventricular hypertrophy and improvement of coronary flow reserve]. AB - OBJECTIVE: To elucidate the relationship between regression of hypertensive left ventricular hypertrophy (LVH) and changes of coronary flow reserve (CFR). METHODS: Ninety-six essential hypertensive patients with LVH were randomly divided into 3 groups: ramipril group, losartan group, and combination group. Before the treatment and 6 months after the treatment, left ventricular mass (LVM) was calculated by three-dimensional echocardiography and CFR was evaluated by transesophageal echocardiography with left anterior descending artery. CFR was calculated as the ratio of coronary flow velocity, after intravenous injection of dipyridamole (D), to rest peak velocity (R). All the indexes of the CFR were corrected by LVM. RESULTS: (1) The systolic blood pressure (SBP), diastolic blood pressure (DBP) and LVM were significantly decreased in ramipril group, losartan group and combination group after 6 months treatment (all P < 0.01). Diastolic flow velocity integral was used as one representative index of CFR. Compared with baseline values, the diastolic flow velocity intergral corrected by LVM (D/R DVi(C)) was significantly increased in ramipril group, losartan group and combination group (1.83 +/- 0.61 vs 1.57 +/- 0.58, P < 0.05; 1.94 +/- 0.45 vs 1.53 +/- 0.64, P < 0.01; 2.03 +/- 0.38 vs 1.49 +/- 0.34, P < 0.01). (3) The changes of D/R DVi(C) showed a positive correlation with the changes of LVM (r = 0.579, P < 0.05) and no significant correlation with the decrease of blood pressure (r = 0.288 and 0.295,both P > 0.05). CONCLUSION: Ramipril, losartan and combination of these two drugs all reduce blood pressure and LVH, and increases CFR. CFR corrected by LVM may help assess drug's effect on CFR. Improvement of CFR is associated with the regression of hypertensive LVH. PMID- 12887824 TI - [Relationship between left ventricular diastolic function and plasma brain natriuretic peptide concentration during severe pregnancy-induced hypertension syndrome]. AB - OBJECTIVE: To evaluate the relationship between the left ventricular diastolic function and plasma level of plasma brain natriuretic peptide (BNP) among patients with pregnancy-induced hypertension (PIH) syndrome. METHODS: The left ventricular diastolic function was evaluated by Doppler echocardiography, and plasma BNP level was tested by radioimmunoassay among 36 pregnant women with severe PIH, 32 normal pregnant women, and 21 normal non-pregnant women of childbearing age. RESULTS: The parameters such as peak mitral flow velocity in early diastole (E), E-wave velocity-time integral (EVTI), the ratio of peak mitral flow velocity in early diastole and peak A-wave velocity in late diastole (E/A), peak pulmonary venous diastolic forward flow velocity (D) and D-wave velocity-time integral (DVTI) of the 36 severe PIH patients were significantly lower than those of the 32 normal pregnant women and those of the 21 normal non pregnant women of child-bearing age. But the parameters such as A, A-wave velocity integral (AVTI), and peak pulmonary venous diastolic forward flow velocity (AR) were significantly higher than those of the normal pregnant women and those of the normal non-pregnant women of child-bearing age. E/A ratio, D and DVTI of the normal pregnant women were significantly lower than those of the normal non-pregnant women of child-bearing age, however, A, AVTI, S/D, and AR were significantly higher than those of the normal non-pregnant women of child bearing age. The BNP concentration of the normal pregnant women was significantly higher than that of the normal non-pregnant women of childbearing age, but significantly lower than that of the severe PIH patients. There were significant correlations between left ventricular diastolic function variables (E/A ratio and AR) and BNP concentration in normal pregnant women and in PIH patients. CONCLUSION: The left ventricular diastolic function is slightly damaged in normal pregnant women and significantly damaged in patients with severe PIH. The plasma BNP level of pregnant women can become an excellent index to predict their left ventricular diastolic function. PMID- 12887825 TI - [Expression of leptin and leptin receptor system in woman reproductive organs]. AB - OBJECTIVE: To explore the expression of leptin, leptin receptor (Ob-R) and related signaling proteins in woman reproductive organs. METHODS: Thirty-nine women, aged 27 - 37, with normal menstrual cycle of which 24 being in proliferative phase and 15 in secretive phase, underwent uterine curettage, to obtain the endometrium. The uterine cervix, vaginal wall, and oviducts were resected from 3 women with hysteromyoma. The expression of mRNA of leptin and long isoform of leptin receptor (Ob-Rb) was detected by in situ hybridization. The expression of leptin and Ob-Rb proteins was examined by immunohistochemical staining. Immunohistochemical staining was used to study the expression of the leptin-related signaling proteins: phosphated signal transducers and activator of transcription (p-STAT3), suppressor of cytokine signaling 3 (SOCS3), and protein inhibitor of activated STAT3 (PIAS3). RESULTS: The mRNA and protein of leptin and the mRNA and protein of Ob-Rb were expressed in the glands and mesenchyma of endometrium. The expression of Ob-Rb protein was significantly stronger in secretive phase than in proliferative phase (P < 0.01). No significant difference was found between the expression of leptin protein in proliferative phase and that in secretive phase. In the endothelium SOCS3 and PIAS3 were expressed, however, p-STAT3 protein was only weakly expressed or not expressed. Leptin protein and Ob-Rb protein were expressed in the uterine cervix, vagina wall, and epithelium of oviduct, including glands. However, leptin mRNA and Ob-Rb mRNA were only expressed in the ciliated epithelium of oviduct. CONCLUSION: Leptin may have an effect on women's reproductive function through leptin receptor systems. PMID- 12887826 TI - [Cloning and biological activities of riboenzymes against major histocompatibility complex class II transactivator]. AB - OBJECTIVE: To investigate the cloning and biological activities of riboenzymes against major histocompatibility complex (MHC) class II transactivator (CIITA) so as to explore the feasibility of using hammerhead riboenzyme to create immune tolerance. METHODS: Three riboenzymes against MHC CIITA were synthesized and their activities were evaluated in vitro. Rz464, the riboenzyme with a better digestion effect, was inserted into the vector pIRES2-EGFP (then called pRz464). Human B lymphoma cells of Daudi line were cultured and transfected with pRz464 (then called pRz464-D). Cultured Daudi cells transfected with riboenzyme without the ability against CIITA (Rz-D) were used as control group. The expressions of classic MHC CIITA (HLA-DR, DP, and DQ) on the surface of Daudi cells before and after transfection were examined by flow cytometry. RESULTS: The expressions of HLA-DR, DP, and DQ on the surface of cultured Daudi cells (Rz-D) were 97.9 +/- 0.8%, 94.3 +/- 1.1%, and 54.4 +/- 3.1% respectively. The expressions of HLA-DR, DP, and DQ were inhibited by 88.4%, 83.5%, and 93.4% respectively on the surface of pRz464-D cells. CONCLUSION: The hammerhead riboenzyme Rz464 inhibits the expression of CIITA mRNAs, thus inhibiting the expression of MHC II molecules. It may be used in antigen-specific tolerance induction in allo-transplantation. PMID- 12887827 TI - [Escherichia coli O157:H7 strains carrying Shiga toxin 2 variant slt2vha were isolated from Xuzhou City of Jiangsu Province]. AB - OBJECTIVE: To monitor the changes of predominant Shiga toxin types of Escherichia coli O157:H7 in Xuzhou City, Jiangsu Province. METHODS: PCR typing with slt2 specific primer pairs (slt2cslt2, slt2v1slt2v2), hybridization of chromosome DNA digested by PstI, and DNA sequencing of PCR products. with slt2-specific primer pairs (slt2cslt2d; slt2eslt2f) were conducted on 14 strains of E. coli isolated from diarrhea patients and dung beetles in 1999 and 2000. RESULTS: The 5 strains of E. coli isolated from diarrhea patients in 1999 carried slt1 and slt2. The 1 strains of E. coli O157:H7 isolated from diarrhea patients and the 5 strains isolated from dung beetles in 2000 only carried slt2vha, a variance of Shiga toxin 2. CONCLUSION: The predominant Shiga toxin types of E. coli O157:H7 in Xuzhou changed 1999 - 2000, which may be related to the changes of epidemiological features of E. coli O157:H7 infection in Xuzhou. A genotyping method based on restriction fragment length polymorphism (RFLP) analysis of a B subunit-encoding DNA fragment obtained by PCR described in this study is a useful tool to identify slt2 gene and slt2 variant gene in epidemiological studies with O157:H7 strains. PMID- 12887828 TI - [The study of cis-element HNF4 in the regulation of mfg12 prothrombinase/fibroleukin gene expression in response to nucleocapsid protein of MHV-3]. AB - OBJECTIVE: To identify the transcription factor(s) that is essential for activation of mfgl2 prothrombinase/fibroleukin gene in response to nucleocapsid protein of murine hepatitis virus type 3 (MHV-3). METHODS: Western blotting was performed to investigate whether HNF4 is expressed in macrophages of Ba1b/c mice where mfgl2 is expressed. Confocus microscope immunofluorescence was performed to show whether N protein of MHV enters into the nucleus of infected cells, which is a critical step for the N protein to facilitate its transactivation property. To facilitate the identification of three candidate factor(s) including hepatocyte nuclear factor 4 (HNF4)/liver factor A1 (LF-A1), cytomegalovirus immediate early gene 1.2 (IE1.2) regulatory element and granulocyte- macrophage colony stimulating factor (GM-CSF) in response to mfgl2 activation upon the stimulation of MHV-A59 N protein, gel mobility shift assay (GMSA), competition experiments and site directed mutagenesis were performed. RESULTS: Western blotting displayed that HNF4 was constitutively expressed in macrophages and did not show significant change under the stimulation of different MHV. Confocus microscope immunofluorescence clearly showed that N protein of MHV entered into the nucleus of infected cells. GMSA and competition experiments demonstrated binding to both HNF4 and IE1.2 fragments could be competed with the cold specific oligonucleotides but not with the same amount of non-specific oligos nucleotides. A super shift band was observed when HNF4 antibody was pre-incubated with the nuclear extracts indicating the interaction between the HNF4 element and mfgl2 promoter. Site directed mutagenesis of cis-elements HNF4 (pfgl2HNF4mut) and HNF4/IE1.2 (pfgl2HNF4/IE1.2mut) mutations abolished over 75% of transcription from wild-type mfgl2 promoter. However the pfgl2IE1.2mut displayed almost wild type promoter activity (75% approximately 80%). CONCLUSIONS: The factor HNF4 binds to mfgl2 promoter and serves as an essential transcription factor for mfgl2/fibroleukin expression in response to MHV-3 N protein. PMID- 12887829 TI - [Collectrin, a kidney-specific novel gene, is over-expressed in the hypertrophic phase in 5/6 nephrectomized rat kidney]. AB - OBJECTIVE: To investigate the expression of the novel gene, collectrin, in the remnant kidney of the 5/6 nephrectomized rat. METHODS: Thirty-four rats were randomly assigned to three groups: sham-operation group (CON, n = 10), 5/6 nephrectomy group (NX, n = 12), NX + enalapril group (ACEI, since one day before the 5/6 nephrectomy 10 mg x kg(-1) x d(-1) enalapril was administered). Two weeks, 8 weeks, and 15 weeks after the above-mentioned treatments, 1/3 of the rats in different group were killed and their kidneys were taken out. Immunohistochemical staining was used to detect the expression of collectrin. The collectrin mRNA expression in the kidneys was detected by RT-PCR, and collectrin protein expression was observed by Western blot assay. RESULTS: In the kidneys of the CON group rats, the expression of collectrin was progressively decreased over time (2 W vs 16 W, 1.69 +/- 0.09 vs 1.21 +/- 0.09, P < 0.05; 8 W vs 16 W, 1.38 +/ 0.04 vs 1.21 +/- 0.09, P < 0.05). However, collectrin was over-expressed in the remnant renal tissues compared with in the kidneys of control rats (NX vs CON, 2 W 2.33 +/- 0.07 vs 1.69 +/- 0.09, P < 0.05; 8 W 1.85 +/- 0.20 vs 1.38 +/- 0.04, P < 0.05; 16 W 1.47 +/- 0.17 vs 1.21 +/- 0.09, P < 0.05); In NX group, collectrin expression was obviously increased 2 weeks after the operation, then decreased along with the progression of renal failure (2 W vs 8 W, 2.33 +/- 0.07 vs 1.85 +/ 0.20, P < 0.05; 2W vs 16 W, 2.33 +/- 0.07 vs 1.47 +/- 0.17, P < 0.05; 8 W vs 16 W, 1.85 +/- 0.20 vs 1.47 +/- 0.17, P < 0.05). In ACEI treated group, collectrin expression was decreased along with the amelioration of renal lesions. The expression of collectrin mRNA was consistent with the expression of collectrin protein. CONCLUSION: The expression of collectrin is progressively decreased with aging in normal rat kidney. In 5/6 nephrectomized kidney the expression of collectrin is related with the renal proliferation and hypertrophy. PMID- 12887830 TI - [Role of nuclear factor of kB in tumor necrosis factor alpha-mediated INS-1 cell apoptosis]. AB - OBJECTIVE: To investigate the effect of cytokine tumor necrosis factor alpha(TNFalpha) on pancreatic beta cells apoptosis and the effect of transcription factor nuclear factor of kappa B (NF-kB) on TNFalpha-mediated apoptosis. METHODS: INS-1 cells, a pancreatic beta cell line, were cultured,and recombinant DNA technique, transfection and reinfection technology were used to obtain INS-1/IkBDeltaN cells expressing inhibitor of kB (IkB)alphaDeltaN, mutant IkBalpha (inhibitor of NF-kB). DNA fragmentation assay and fluorescence analysis using a kB-luc reporter gene were applied to observe the NF-kB activity and beta cell apoptosis. RESULTS: NF-kB activity induced by IL-Ibeta was detectable in INS 1 cells but not in INS-1/IkBDeltaN cells. After incubation of the cells with IL 1beta (10 micro g/L), TNFalpha (100 micro g/L) and interferon (IFN)gamma (100 000 U/L) for 48 hours, the combination of TNFalpha and IFNgamma induced apoptosis in INS-1 cells but not in INS-1/IkBDeltaN cells. No apoptosis was observed after incubation of INS-1 cells with IL-1beta or IFNgamma or IL-1beta plus IFNgamma. CONCLUSION: Apoptosis is one of the TNFalpha-induced beta-cell death forms. NF-kB may play an important role in the TNFalpha-mediated beta-cell apoptosis. Inhibition of NF-kB activation protects beta-cells from TNFalpha-induced apoptosis. PMID- 12887832 TI - New Milestones for JCPSP. PMID- 12887831 TI - [A low level of TNF-mediates hemorrhage-induced acute lung injury via p55 TNF receptor]. AB - OBJECTIVE: To examine the temporal relationship of pulmonary TNF-alpha production to acute lung injury (ALI) during hemorrhagic shock (HS). METHODS: HS was induced in mice by removal of 30% of calculated total blood volume. Lung TNF-alpha was measured by ELISA. Lung neutrophil accumulation was detected by immunofluorescent staining, and pulmonary microvascular permeability was assessed using Evans blue dye. RESULTS: While HS induced a slight and transient increase in lung TNF-alpha, neutrophil accumulation preceded the change in lung TNF-alpha. However, lung neutrophil accumulation and the increase in microvascular permeability were abrogated in TNF-alpha knockout mice, and both were restored by administration of low dose TNF-alpha to TNF-alpha knockout mice prior to HS. Both neutrophil accumulation and microvascular leak were abrogated in p55 TNF-alpha receptor knockout mice, while p75 TNF-alpha receptor knockout mice behaved similar to wild type. CONCLUSION: A low level of pulmonary TNF-alpha is sufficient to mediate HS induced acute lung injury and that the p55 TNF-alpha receptor plays a dominant role in regulating the pulmonary inflammatory response to HS. The results suggest that anti-TNF-alpha strategies for the control of the pulmonary inflammatory response to HS can be directed toward antagonizing the p55 TNF-alpha receptor. PMID- 12887833 TI - Anesthesia related complications of laparoscopic cholecystectomy. AB - OBJECTIVE: To determine the incidence of intraoperative anesthesia-related complications of laparoscopic cholecystectomy. DESIGN: A descriptive study. PLACE AND DURATION OF STUDY: The study was conducted in Anesthesia Department of Hayat Shaheed Teaching Hospital, Peshawar from April 1995 to April 1996. SUBJECTS AND METHODS: One hundred consecutive patients (ASA I and ASA II) were anesthetized and monitored intraoperatively for detection of complications. The operative findings and data were recorded on a proforma and finally the results were analyzed. RESULTS: One hundred patients with male to female ratio of 1: 8.09 in the age range of 20-80 years (mean 39 years) underwent general anesthesia for laparoscopic cholecystectomy. The duration of operation in 94 laparoscopic cholecystectomy was from 20 to 80 minutes (mean 60.63 minutes ).The incidence of intraoperative hypotension was 9%. Four percent of the patients developed arrhythmias. Increase in end-tidal-carbon dioxide (ETCO2) was observed in 3% of cases. Conversion rate to open cholecystectomy was 6%. Damage to intraabdominal vessels with trocar insertion occurred in 1% of cases. CONCLUSION: Although laparoscopic cholecystectomy has major surgical and anesthetic advantages, there are anesthesia related complications requiring specific anesthetic interventions to improve patients outcome without compromising their safety. PMID- 12887834 TI - Autotransplantation of endodontically treated third molars. AB - OBJECTIVE: To determine the success rate in autotransplantation of endodontically treated third molars. DESIGN: A descriptive study. PLACE AND DURATION OF STUDY: This study was carried out at Armed Forces Institute of Dentistry (AFID), Rawalpindi (Pakistan) from January 2002 to December 2002. PATIENTS AND METHODS: A total of 50 patients meeting the inclusion criteria were selected who had their first or second molars in unrestorable condition with intact third molars. The donor teeth were extracted after the preparation of recipient site. After endodontically treated in vitro the donor teeth were carried to the recipient site and immobilized. Postoperative variables were recorded and analyzed on SPSS version 10. RESULTS: The overall success rate after six months of the transplantation of third molars was 88% with complete root formation after endodontic treatment. All the patients(12%) who had complaints were more than 35 years of age. CONCLUSION: Third molars are good substitute for the unrestorable first or second molars and would be as effective as endosseous implants. The procedure is likely to have complications in the advanced age group. PMID- 12887835 TI - Spectrum of dysfunctional uterine bleeding and its conservative management. AB - OBJECTIVE: To find out the proportion of dysfunctional uterine bleeding (DUB) and the response of patients of DUB to medical treatment at Abbasi Shaheed Hospital. DESIGN: Cross sectional and comparative study. PLACE AND DURATION OF STUDY: The study was conducted in Gynaecology Unit II of Abbasi Shaheed Hospital, Karachi, from December 1999 to December 2000. PATIENTS AND METHODS: The study included 210 Gynae patients between the age group 13-50 years presenting with DUB. These patients were divided into adolescent, reproductive and perimenopausal group. They were initially assessed by general and pelvic examination, ultrasound of pelvis, Pap smear, hysteroscopy and diagnostic curettage. Results were recorded. The efficacy of treatment was assessed in terms of reduction in the amount of blood loss. RESULTS: Two hundred and ten out of 1300 patients were diagnosed as having DUB with the proportion of 16.1%. Response rate was 20 30 % with oral mafenamic acid, 50% with capsule tranexamic acid, 60% and 50% respectively with oral contraceptive pills containing ethinylestradiol and norethisterone, and norethisterone alone. The response rate with capsulated micronised flavonoid and derivative of ethinyltestosterone was 90% and 75% respectively. CONCLUSION: The patients in adolescent age group are good responders to conservative treatment. Most of the patients preferred micronised flavonoid over derivative of ethinyltestosterone for some misconceptions regarding this medicine. The former was better tolerated with less side effects. PMID- 12887836 TI - Introducing problem-based learning in a medical school with traditional/conventional curriculum. AB - BACKGROUND: Globally, over the last two decades, medical education and its learning strategies are being made more prone to problem-solving in order to produce better clinicians. The medical curriculum has not changed in Pakistan since its inception and that is reflected in the form of inadequacies in health delivery system. There is a dire need of creating relevance between medical education/training and medical practice in order to introduce an effective health care system. OBJECTIVE: This study is particularly aimed at evaluating the effect of problem-based learning (PBL) in a group of students accustomed to conventional teaching and to assess any change in students attitudes/learning abilities after repeated PBL interventions. DESIGN: An interventional study. SUBJECTS AND METHODS: Response of 32 students of fourth year MBBS of Karachi Medical and Dental College was recorded against two PBL exercises in September 2002. RESULTS: A significant change in the study trend and analytical ability (p < 003) was shown in the two PBL sessions. The students who liked PBL were 33% more prone to practice group-study (OR = 1.33 and 95% CI=1.28, 1.39) than those who preferred other modalities of teaching. The final professional grades of these students were significantly higher than those in the previous years (p < 0.000). CONCLUSION: A strong association between liking for PBL and group study was found and the analytical performance and thought process of students were significantly improved after the second PBL session. PMID- 12887837 TI - Factors predicting hyperkalemia in patients with cirrhosis receiving spironolactone. AB - OBJECTIVE: To evaluate the factors leading to hyperkalemia in patients with cirrhosis receiving spironolactone. DESIGN: An observational, analytical, case control study. PLACE AND DURATION OF STUDY: The Aga Khan University Hospital. Six months. SUBJECTS AND METHODS: One hundred and fifty consecutive patients (100 males, 50 females) with cirrhosis of liver, hospitalized for decompensated disease and receiving spironolactone for at least two weeks before admission, were included in this study. Patients with hyperkalemia (n=67) having potassium level >5 mmol/l were compared with patients who had normal potassium level 5 mmol/l (n=83). The parameters taken into account were age, gender of the patients, type and dose of diuretics along with concomitant medicines, diastolic blood pressure, edema, ascites, blood urea nitrogen, serum creatinine, electrolytes, bilirubin, albumin, prothrombin time, Child class, and Child Pugh score. RESULTS: Patients with hyperkalemia (K > 5 mmol/l) had higher blood urea nitrogen, serum creatinine and bilirubin levels (p= 0.004, 0.001 and 0.044 respectively). Their serum sodium and albumin levels were lower (p= 0.000 and 0.017 respectively). They had advanced cirrhosis with high Pugh score (p= 0.003). These patients were on higher dose of spironolactone (p =0.001). Multivariate analysis showed that dose of spironolactone >100 mg /day, serum creatinine >1.3 mg/dl, persistence of ascites and edema, and female gender were important predictors of development of hyperkalemia. CONCLUSION: Patients with cirrhosis receiving high dose of the diuretic, having edema, ascites and high serum creatinine are at the greater risk of developing hyperkalemia during spironolactone therapy. PMID- 12887838 TI - Tobacco use as contributory factor in peptic ulcer disease. AB - OBJECTIVE: To find out the association of tobacco as a contributory factor in patients presenting with peptic ulcer disease (PUD). DESIGN: A descriptive case study. PLACE AND DURATION OF STUDY: Department of Medicine, Postgraduate Medical Institute, Lady Reading Hospital, Peshawar, from May 2001 to April 2002. SUBJECTS AND METHODS: A total number of 100 consecutive patients, 76 males and 24 females, with endoscopically proven peptic ulcer were enrolled in the study. The ages of the patients ranged from 25 to 70 years; with mean age of 48 years + 12.26 standard deviations, (mean age of men 49 years + 12.88 SD, and mean age of women 45 years + 9.67 SD). All the patients underwent endoscopy after an overnight fast, using lignocaine (4% solution or jelly) as topical anaesthetic, with Pentax FG29 video-endoscope. RESULTS: Fifty-six percent of all the patients with PUD used tobacco, 33% as cigarette, 18% took it as naswar and 5% of patients used it in both the forms. Of these, 85.7% (48/56) patients had duodenal ulcer and 14.3% (8/56) patients had gastric ulcer. All these patients were male, none of the female patients used tobacco in any form. Moreover, 40% of PUD patients reported long-term use of nonsteroidal anti-inflammatory drugs (NSAIDs). CONCLUSION: This study supports the view that use of tobacco is associated with PUD in men. PMID- 12887839 TI - Significant variables associated with epilepsy. AB - OBJECTIVE: To study the characteristics of the epileptics and the risk factors contributing to the development of epilepsy. DESIGN: Descriptive study. PLACE AND DURATION OF STUDY: Epilepsy Clinic at Ahbab Hospital, Lahore, from June 2002 to August 2002. SUBJECTS AND METHODS: Data was collected from 158 subjects, 89 males and 69 females, suffering from epilepsy. The information about socio-demographic characteristics and family history of illness, perinatal morbidity, birth place and mother s age at the time of delivery was obtained using a pre-tested questionnaire. Data was analyzed on SPSS version 10. RESULTS: Majority of the subjects were single (77.84%), 1st born among their siblings (25.95%), belonged to low social class (50.63%), and unemployed (25.31%). The major risk factors were family history of illness (23.52%) and positive medical problem around birth (12.66%). The presence of family history of illness, positive medical problem around birth and advanced maternal age at birth were associated with early onset of epilepsy. Vulnerability for the epilepsy also increases among hospital deliveries. CONCLUSION: Although the present study has identified various risk factors, yet the results need to be further confirmed through case-control studies. PMID- 12887840 TI - Validaty of Siriraj Stroke Scoring. AB - OBJECTIVE: To study the sensitivity and specificity of Siriraj Stroke Scoring (SSS) and to validate the accuracy of SSS in acute supratentorial stroke syndromes. DESIGN: Noninterventional descriptive study. PLACE AND DURATION OF STUDY: Department of Neurology, Pakistan Institute Of Medical Sciences, Islamabad, and Department of Medicine, Federal Government Services Hospital, Islamabad, from July to December 2000 and February to July 2002, respectively. SUBJECTS AND METHODS: One hundred (100) consecutive cases of acute supratentorial strokes were studied in accordance with SSS and sensitivity and specificity of SSS for supratentorial infarction and haemorrhage was tested against the computerised brain scanning (CT) as a gold standard. Siriraj Stroke Scoring was applied. Patients with subarachnoid haemorrhage and transient ischemic attacks were excluded from the study. The findings were recorded, compared and statistically analyzed. RESULTS: Out of 100 patients 45 were male and 55 female with mean age of 59 years. Sixty-two patients had hypertension, 36 patients had one or more atheroma markers (diabetes, coronary artery disease and claudication). Mean diastolic blood pressure was 98 mmHg with range of 60-140 mmHg and mean systolic blood pressure of 163 mmHg with the range of 90-240 mmHg. Forty-eight patients had cerebral infarction, 36 had cerebral hemorrhage while 16 were borderline cases on the basis of Siriraj stroke scoring. CT brain showed 56 and 39 patients had cerebral infarction and haemorrhage respectively while 5 of CT scans were normal. The sensitivity and specificity of SSS for cerebral infarction was 71 and 85 respectively and for intracerebral haemorrhage, it was 73 and 90 respectively. The positive predictive values of SSS for cerebral infarction and haemorrhage were 87% and 83 respectively. CONCLUSION: The siriraj stroke scoring system is a valid and specific scoring system for the diagnosis of acute supratentorial stroke and intracerebral haemorrhage. PMID- 12887841 TI - Corneal complications of vernal catarrh. AB - OBJECTIVE: To identify the magnitude and types of corneal complications of vernal catarrh and the extent of visual impairment caused by them. DESIGN: A cross sectional study. PLACE AND DURATION OF STUDY: Khyber Institute of Ophthalmic Medical Sciences (KIOMS), Hayatabad Medical Complex (HMC), Peshawar, from March, 1999 to May, 2000. SUBJECTS AND METHODS: All cases of vernal catarrh who agreed to an informed consent were included in the study. Patients who were difficult to be examined by a slit lamp were excluded from the study. The diagnosis was made mainly on the basis of history and clinical examination. Slit lamp examination was done in every case together with fluorescein staining of cornea. The variables were recorded and statistically analyzed. RESULTS: Superficial punctate keratitis (45%) was the most common corneal complication. Visual impairment was more pronounced in cases with shield ulcers (14%) and corneal plaques (8%). A strong association with keratoconus (15%) was also noted. Corneal opacification (9%), hydrops (6%) and pseudogerentoxon (3%) were also found. CONCLUSION: Corneal complications in VKC are common, potentially serious and may cause marked visual impairment. PMID- 12887842 TI - Hyphaema due to blunt trauma. AB - OBJECTIVE: To study the causes, clinical presentation, complications and visual outcome of hyphaema following closed globe injury. DESIGN: Prospective study. PLACE AND DURATION OF STUDY: The study was conducted at the Department of Ophthalmology, Postgraduate Medical Institute, Lady Reading Hospital, Peshawar, Pakistan, from May, 1996 to June 1997. PATIENTS AND METHODS: All cases of hyphaema, due to closed globe injury, were included and history of patients was recorded on a comprehensive proforma designed for this purpose. All cases were admitted for at least 5 days. Ocular examination included checking visual acuity (VA), intraocular pressure (IOP), slit lamp and fundus examination. Patients were treated accordingly and followed up for 90 days. RESULTS: Thirty-six eyes of 36 patients with hyphaema were included in our study. Out of this, 88.89% were males and 11.11% females while 86% were 20 years of age or below. Almost 91.66% presented with primary hyphaema, 44.44% received trauma with stone and 27.77% with tennis ball while playing cricket. Trauma during playing was noted in 66.66% of cases. Almost 62% of cases were having visual acuity in the range of CF to perception of light with good projection. IOP was raised initially in 41.66% cases. Only 33.33% needed surgical intervention. Secondary glaucoma was present in 13.88% and 5.55% had corneal blood staining. Majority of patients (75%) improved to VA range of 6/6 to 6/12 and 19% patients in our study ended up with legal blindness in the affected eyes. CONCLUSION: Traumatic hyphaema is more frequently found among children and young male. Angle recession, traumatic cataract and maculopathy were common co-morbidities. Secondary glaucoma and corneal staining were the main complications. Nearly 1/5th of patients ended up with legal blindness in the affected eyes. PMID- 12887843 TI - Comparison of response between food supplemented with powdered iron and iron in syrup form for iron deficiency anemia. AB - OBJECTIVE: To evaluate and compare the response between food supplemented with iron in powdered and iron in syrup forms for the treatment of iron deficiency anemia in children aged 1-5 years. DESIGN: Quasi-experimental study. PLACE AND DURATION OF STUDY: This study was conducted at Combined Military Hospital, Multan, from January 2000 to December 2000. PATIENTS AND METHODS: Consecutive 200 cases of iron deficiency anemia, aged 1-5 years, were included in the study. Diagnosis was based on history, physical examination, complete blood counts (CBC) and serum ferritin levels. The patients were randomized to receive either iron in syrup form (Group A) or equivalent doses of iron powder sprinkled over food (Group B). The patients were followed up with hemoglobin estimation (Hb) and reticulocyte response at 2 weeks, 4 weeks and 6 weeks. CBC and serum ferritin were repeated at 6 weeks. RESULTS: Over half (51%) of the patients were between 1 2 years of age. One hundred thirty-two were males and 68 females. Most of the patients belonged to poor socioeconomic class. The iron in powder form was better tolerated than iron syrup as this group witnessed fewer episodes of gastrointestinal disturbances. The rise in mean Hb level after 6 weeks of treatment in group A and B was 1.6 g/dl and 1.9 g/dl respectively. Hemoglobin rise in group B was more than group A but this was statistically non-significant (p > 0.05). There was small but significant (p < 0.05) rise in serum ferritin in both the groups. There was no significant difference between the two groups for response to the two forms of iron administration. CONCLUSION: The powdered form of iron is a cost-effective and better tolerated method of iron administration in children and can be considered as an alternate option for the treatment of iron deficiency anemia in children. PMID- 12887844 TI - Reconstruction of postleishmaniasis scar with lateral cheek rotation flap. AB - OBJECTIVE: the aim of this study was to find a simple and economical technique to remove postleishmaniasis scar from the face. DESIGN: a non-randomized descriptive case series. PLACE AND DURATION OF STUDY: the study was conducted between 1st January 1998 to 31st July 2000 in a private hospital of D.I.Khan. PATIENTS AND METHODS: Ten patients with postleishmaniasis scar on the face were selected for excision and reconstruction in this study. They were operated as day care under local anesthesia. Their age ranged between 18-30 years, with an average of 23.5 years. The size of scar varied from 2 cm2 to 6 cm2 with an average of 4.1 cm(2). After scar excision, lateral cheek rotation flap (reverse face-lift) was used to reconstruct the defect. Results were statistically analyzed. RESULTS: No major complications were encountered. However, minor complications like edema of the flap, seroma under the flap, flap tip necrosis and temporary neurapraxia were noted in two patients. CONCLUSION: This is an excellent technique, which can be done on a day care basis under local anesthesia. Larger studies will help to explore this technique further. PMID- 12887845 TI - Accuracy of magnetic resonance imaging in the knee. AB - OBJECTIVE: To determine the accuracy of magnetic resonance imaging (MRI) in the assessment of various knee disorders in comparison with arthroscopic findings and pathologic diagnosis. DESIGN: A retrospective and comparative study. PLACE AND DURATION OF STUDY: Radiology Department, Aga Khan University Hospital, Karachi, from 01-12-1996 to 30-06-2000. PATIENTS AND METHODS: Fifty-six subjects (36 males and 20 females), included in this study, had abnormal findings on MRI, which were correlated with arthroscopic findings in 50 cases and pathological diagnosis in 6 cases. Plain X-rays were available in all cases. RESULTS: The sensitivity, specificity and accuracy for MRI of the menisci and cruciate ligaments were as follows: medial meniscus pathologies 87% sensitivity, 94% specificity and 92% accuracy; lateral meniscus pathologies 92% sensitivity, 89% specificity and 90% accuracy; anterior cruciate ligament injuries 95% sensitivity, 96% specificity and 96% accuracy and posterior cruciate ligament injuries 100% sensitivity, 98% specificity and 98% accuracy. Pathological diagnosis was available in 6 cases. MR imaging suggested the diagnosis of tuberculosis and metastases as well as mapped out the extent of osteogenic sarcoma, fibromatoses and rhabdomyosarcoma. CONCLUSION: This study confirms that MR imaging of the knee is highly sensitive, specific, and accurate. PMID- 12887846 TI - Outcome of primary repair in penetrating colonic injuries. AB - OBJECTIVE: To assess the outcome of primary repair in penetrating colonic injuries in terms of regional/local morbidity (suture line breakdown, intra abdominal abscess) and mortality. DESIGN: Prospective, interventional study. PLACE AND DURATION OF STUDY: Surgical Unit II at Lahore General Hospital, Lahore, over a period of 3 1/2 years from June 1999 to December 2002. PATIENTS AND METHODS: Out of 38 consecutive patients with penetrating colon injuries, a selective group of 25 patients (65.7%) undergoing primary repair (simple suture, resection and anastomosis without covering colostomy and right hemicolectomy) were included in this study. The morbidity and mortality variables were recorded and statistically analyzed. RESULTS: Majority of our patients were of younger age group (mean 25 years). Twenty patients (80%) were male. The commonest mode of injury was firearm injury (72%). The time interval between injury and repair was 3-11 hours (mean 7 hours). Simple repair of colon with interrupted stitches was the commonest procedure performed (44%), followed by right hemicolectomy (32%). Colon related complications developed in 3 patients (12%), which included two fecal fistulae and one intra-abdominal collection. One patient died of septicemia (4%). Hospital stay ranged between 6-16 days (mean 8 days). CONCLUSION: Primary repair is a safe method of managing penetrating colon injuries in carefully selected patients. PMID- 12887847 TI - Dermoid cyst of the floor of mouth. AB - A case report of a 30-year-old male with a huge suprahyoid neck swelling and causing respiratory as well as oral function difficulties is reported. The mass was enucleated through oral route, following which normal oral functions as well as respiratory ease were restored. The gross appearance was consistent with dermoid form of congenital cyst of the mouth floor. PMID- 12887848 TI - Liver cyst with inflammatory hepatic duct polyp. AB - This is a case report of a 50-year-old woman, who presented with upper abdominal pain, vomiting and two episodes of jaundice in five months. Computerised tomographic scan revealed a cyst in the 4th segment of the liver. Peroperatively it was found to be a simple liver cyst arising from the left hepatic duct. The cyst was excised. Peroperative cholangiogram revealed obstruction in the common hepatic duct. A polyp in the common hepatic duct, causing obstruction, was excised. The presence of liver cyst with hepatic duct polyp is a rare association. PMID- 12887849 TI - Physical exercise as a key health determinant among patients. PMID- 12887850 TI - Brain death and related issues. AB - Concerns about the erroneous diagnosis of death and premature burial have been expressed from times immemorial. Patients with brain stem death have absolutely no chance of recovery. brain death is considered at par with death in most of the countries. General public in most parts of the world shows reluctance to accept this concept due to different social, cultural and religious backgrounds and state of literacy and awareness. The criteria for the diagnosis of brain death have been established which include certain pre-conditions, exclusions and tests of the brain stem function. These criteria are universally accepted. The criteria in children are somewhat different from the adults. The subject is intimately related with organ transplantation. If the patient is registered as organ donor or the family consents, organs can be harvested from brain dead patients for transplantation. Pakistan is amongst the few countries where no legislation exists to accept brain death as being at par with death of an individual, and to facilitate and regulate, cadaveric organ donation and transplantation. PMID- 12887851 TI - Vancomycin-resistance Staphylococcus aureus. PMID- 12887852 TI - [Distribution of polyps in the inside region of Vizcaya (Spain): implications for diagnostic tests and colorectal cancer screening]. AB - INTRODUCTION: The importance of colonic polyp is increasing because colorectal cancer (CRC) screening is based on their identification. However, consensus is lacking on which technique should be used in CRC screening, whether colonoscopy or other methods such as sigmoidoscopy, which shows good sensitivity only in distal segments of the colon. We studied the characteristics and localization of polyps in our environment to verify their tendency to change localization and determine the implications of this in screening. MATERIAL AND METHODS: We reviewed the prevalence and characteristics of colonic polyps in 3604 patients who underwent colonoscopy for various causes in our service, as well as their distribution in 837 patients with a diagnosis of polyps who underwent complete colonoscopy. Localization proximal or distal to the splenic flexure and polyp size (less than or more than 1 cm) were analyzed. RESULTS: Polyps were found in 28.3% of 3604 patients (11.7% showed polyps > or = 1 cm). Of the 837 patients who underwent complete colonoscopy, proximal polyps were found in 44.9% (14.6% showed polyps > or = 1 cm), while distal polyps were found in 79.9% (30.3% showed polyps > or = 1 cm). Both proximal and distal polyps were found in 24.8%. The probability of finding both proximal and distal lesions was 31%. Proximal lesions without distal lesions were found in 20% of the patients (5.6% presented polyps > or = 1 cm), and consequently sigmoidoscopy would miss 15.6% of polyps > or = 1 cm. CONCLUSIONS: A relatively high percentage of patients with proximal polyps > or = 1 cm would remain undiagnosed if complete colonoscopy were only performed in patients with distal polyps. Because of the well-known increase in the frequency of proximal polyps and because a considerable number of these present without distal sentinel lesions, there are increasing arguments for considering colonoscopy as the basic technique for CRC screening. PMID- 12887853 TI - [Genetic variability of hepatitis C virus in the health area of Elche (Spain). Correlation between core antigen and viral load]. AB - We investigated the prevalence of the various genotypes of hepatitis C virus (HCV) in 281 patients evaluated between March, 2000 and March, 2002 in the health area of Elche. Of these patients, 55 were coinfected with human immunodeficiency virus (HIV). The genotype was related to viral load and the co-existence of HIV infection. Likewise, the relationship between these parameters and the presence of the HCV core antigen was established. The results indicate that genotype 1b was the most prevalent (38.4%) followed by genotype 3a (23.1%). Patients coinfected with HIV presented fewer infections due to group 1 genotypes (p < 0.05).Patients with HIV presented a greater viral load in all the genotypes, with genotype 3 presenting a high viral load. Detection of the HCV core antigen showed a close correlation with viral load determinations. Although not yet sufficiently assessed, determination of the HCV core antigen constitutes a simple technique that could eventually contribute to improving the management of patients with chronic HCV hepatitis. PMID- 12887854 TI - [Relationship between patient's subjective stress perception and the course of inflammatory bowel disease]. AB - INTRODUCTION: Studies examining the relationship between stress secondary to adverse life events (ALE) and inflammatory bowel disease (IBD) have produced controversial data. The aim of this study was to explore the subjective perception of this relationship in IBD patients. PATIENTS AND METHODS: Seventy consecutive patients suffering from IBD (40 Crohn's disease, 30 ulcerative colitis) were assessed using a self-rating questionnaire related to demographic variables, clinical characteristics, subjective perception of the influence of ALE on the course of IBD, psychiatric background, and the HAD scale. The results of this scale were compared with a group of 25 relatives. RESULTS: Forty-two patients (60%) perceived that there was relationship between ALE and the onset of their disease, forty-nine (70%) between ALE and the increasing IBD symptoms severity, and fifty-one patients (72.9%) with disease activity. Sixteen of the patients (22.9%) had been visited by a psychiatrist during relapses of IBD. Twenty-five patients (42.4%) reached a score of 11 or higher on the depression or anxiety subscales of the HAD, indicating a probable psychological disorder. CONCLUSIONS: IBD patients perceive a strong relationship between ALE and the course of IBD. We have found a high prevalence of anxiety and depression symptoms in these patients. If this observation is confirmed with objective measurements, it will be important to consider psychiatric intervention for these patients. PMID- 12887855 TI - [Liver failure due to mushroom poisoning: clinical course and new treatment perspectives]. AB - Mushroom poisoning, mainly due to the Amanita genus, is an infrequent cause of liver failure in our environment. However, because of its high morbidity and mortality, it constitutes a medical emergency. The characteristic initial symptoms of vomiting, abdominal pain, and diarrhea are nonspecific and may be confused with gastroenteritis. If correct and early treatment is not given, renal and hepatic failure can develop, sometimes requiring liver transplantation. We present three cases of mushroom poisoning, which presented a different clinical course ranging from complete recovery with traditional medical treatment to severe acute liver failure requiring transplantation in one patient and albumin dialysis (molecular absorbent recycling system [MARS]) in another with favorable outcome. Although controlled clinical studies of the treatment of mushroom poisoning are lacking, recommendations based on the experience of various authors have been established. Penicillin G and silymarin seem to be useful. The development of new techniques of extracorporeal detoxification, mainly MARS, may represent an important support system in the treatment of these patients. PMID- 12887856 TI - [Ursodeoxycholic acid treatment shortens the course of cholestasis in two patients with benign recurrent intrahepatic cholestasis]. AB - INTRODUCTION: Benign recurrent intrahepatic cholestasis (BRIC) is a rare autosomal-crecessive or sporadic disorder, characterized by repeated episodes of unexplained cholestasis followed by prolonged asymptomatic periods. CASE REPORT: We present two male patients aged 12 and 15 years old who initially presented cholestasis and who had no family history of liver or biliary disease. Typically, alkaline phosphatase and bilirubin levels were elevated, with a slight increase in transaminases and gamma-glutamyltransferase. In both patients biliary tract disease was ruled out by endoscopic retrograde cholangiopancreatography and magnetic resonance cholangiography while no significant abnormalities of the liver parenchyma were found on liver biopsy. Early treatment with ursodeoxycholic acid (UDCA) reduced the duration of the cholestasis episode compared with that of other episodes in which the patient had received other treatment or the treatment was late (1 or 2 months vs 4 months in patient 1 and 1-3 months vs 5 months in patient 2, respectively). DISCUSSION: Consensus is lacking on the treatment of BRIC although several treatment have been used. UDCA could be effective in increasing bile acid secretion and in reducing serum bilirubin levels. In the two patients described, the initial dosage was 15 mg/kg/day, which was subsequently maintained at 6 mg/kg/day, which was subsequently maintained 6 mg/kg/day until biochemical parameters returned to normal. Nevertheless, larger studies are required to confirm that UDA shortens episodes of cholestasis in BRIC. PMID- 12887857 TI - [Cholestatic jaundice and constitutional syndrome as early manifestations of primary systemic amyloidosis]. AB - We present the case of a 70-year-old woman who had been suffering from constitutional syndrome for several months, abdominal distension, and yellowish coloration of the skin for the previous few days with a rapidly fatal course. Examination revealed hepatomegaly and ascites. Laboratory investigations revealed hyperbilirubinemia with cholestasis. The remaining investigations (abdominal ultrasound, barium transit evaluation, bone marrow study, analysis of ascitic fluid and laparoscopy) did not establish the diagnosis. This was established by liver and subcutaneous fatty tissue biopsies, which revealed type AL amyloid deposits. Autopsy confirmed that the patient had primary systemic amyloidosis. This infrequent form of presentation of systemic amyloidosis and its poor prognosis are discussed. PMID- 12887858 TI - [Training of specialists in gastroenterology. Was it time for a review?]. PMID- 12887859 TI - [New horizons in the mechanisms of acute and chronic pancreatic damage]. PMID- 12887860 TI - [Circulatory dysfunction in cirrhosis. Physiopathology and clinical implications]. PMID- 12887861 TI - [Peritoneal tuberculosis with poor outcome]. PMID- 12887862 TI - [Prognostic significance of the mucin profile in colon cancer without lymph node metastases A, B1 and B2]. PMID- 12887863 TI - [Rhomboencephalitis due to Listeria monocytogenes as a complication of Crohn's disease]. PMID- 12887864 TI - [Mucinous cystoadenoma and recurrent pancreatitis]. PMID- 12887865 TI - [Presacral cyst and serum CA 19.9 raising]. PMID- 12887866 TI - [Fetuses, neonates and corticoids]. PMID- 12887867 TI - [Chronic and recurrent immune thrombocytopenic purpura]. AB - BACKGROUND: Immune thrombocytopenic purpura (ITP) is characterized by a drop in platelet count usually accompanied by hemorrhagic diathesis. In chronic forms the platelet count remains low for six months after diagnosis and in recurrent forms the drop in platelet count appears after a period of normality. OBJECTIVES: To asses outcome and treatment response in patients with chronic or recurrent ITP. METHODS: We performed a retrospective, descriptive study of patients attended in the pediatric hematology outpatient clinic between January 1999 and December 2001. RESULTS: Of 38 patients with chronic ITP, 16 (42 %) presented chronic forms and 22 (58 %) presented recurrent forms. No significant differences were found between the two groups in age, sex, diagnosis, duration of follow-up, previous viral infection, or antiplatelet antibodies. In recurrent forms, the most effective treatment was intravenous immune gamma-globulin (77 % favorable responses) but response time was short (mean: 22.1 weeks). Splenectomy produced complete remission in 63 % of the chronic forms. Good results were obtained in six patients from both groups treated with intravenous anti-D immune globulin. During the study period, 4.5 % of patients with recurrent forms and 31.5 % of those with chronic forms showed spontaneous remission without treatment. CONCLUSIONS: In our experience, the most effective treatment for recurrent forms of ITP was intravenous immune globulin, but none of the treatments achieved long term responses. In chronic forms, splenectomy is an effective alternative when the risk of hemorrhage is high, while a watchful attitude seems to be the best option when this risk is absent. Although the number of patients treated with intravenous anti-D immune globulin was low, good results were achieved. PMID- 12887868 TI - [Crystalline microstructure and microanalysis of gallstones in childhood and adolescence]. AB - OBJECTIVES: To determine the crystalline structure and chemical composition of gallstones in children and adolescents. Design of the studyA simple descriptive study was performed in 21 gallstones from pediatric patients with chronic cholecystitis who underwent elective cholecystectomy. Only two patients had an underlying disease that could explain the presence of gallstones. Crystalline structure was determined by X-ray diffraction and visualization with scanning electron microscopy. Emission and atomic absorption spectroscopy were used to study composition. An X-ray-dispersion energy spectroscope joined to the microscope was used to confirm the crystalline composition of some of the areas examined with the microscope. RESULTS: Bilirubinate and calcium carbonate predominated in the crystalline composition, demonstrating the pigmented appearance of idiopathic gallstones. Cholesterol was a secondary element. Microanalysis confirmed calcium as a fundamental constituent of gallstones irrespective of age. Other elements, such as sodium, potassium, magnesium and manganese, etc., were detected. CONCLUSIONS: The present study confirms the pigmented character of idiopathic gallstones in childhood. The high levels of calcium carbonate and bilirubinate, despite a lack of evidence for any risk factors for the formation of bilirubinate, was striking. Calcium was the most predominant element of the calculi. PMID- 12887869 TI - [Transpyloric enteral nutrition in critically-ill children (I): technic and indications]. AB - OBJECTIVE: To study the utility and efficacy of transpyloric enteral nutrition (TEN) in critically-ill children by analyzing the factors that determine enteral tolerance. PATIENTS AND METHODS: We performed a prospective, observational study between 1994 and 2002 of all critically-ill children admitted to our pediatric intensive care unit who received TEN. The indications for enteral nutrition, type of nutrition and its duration were studied. RESULTS: A total of 286 patients (8.4 % of patients admitted to the PICU in the study period), aged between 3 days and 17 years received TEN. Fifty-five children (19.2 %) were aged less than 1 month and 165 (27.7 %) were aged less than 6 months. Seventy percent received TEN exclusively. The most frequent indication for TEN was mechanical ventilation in 255 children (89.2 %). One hundred seventy-six patients (61.5 %) received TEN in the postoperative period after cardiac surgery. The mean duration of TEN was 15.4 25 days, the maximum volume of nutrition was 118.7 41 ml/kg/day, and the maximum caloric intake was 88.6 26.7 kcal/kg/day. During TEN, 227 patients received sedation (79.3 %), and 124 were administered muscle relaxants (43.3 %), with no increase in complications. CONCLUSIONS: TEN is a useful method of nutrition in critically-ill children. PMID- 12887870 TI - [Transpyloric enteral nutrition in critically-ill children (II): complications]. AB - OBJECTIVE: To study tolerance to transpyloric enteral nutrition (TEN) and the incidence of secondary complications in critically-ill children. PATIENTS AND METHODS: We performed a prospective, observational study between 1994 and 2002 of all critically-ill children admitted to our pediatric intensive care unit who received TEN. Tolerance and complications were analyzed. RESULTS: Of 286 patients aged between 3 days and 17 years who received TEN, gastrointestinal complications occurred in 13.6 %, abdominal distention and/or excessive gastric residue in 8 % and diarrhea in 6.3 %. Diarrhea was associated with shock (p 0.01), abdominal distension and/ or excessive gastric residue (p 0.008), hypophosphatemia (p 0.001), and duration of TEN (p < 0.001). TEN was discontinued in 2.1 % of the patients because of gastrointestinal complications. Thirty-two patients (11.2 %) died during TEN. No relationship was found between the characteristics of nutrition and complications and mortality. CONCLUSIONS: TEN is a well tolerated method of nutrition in critically-ill children that produces few complications. PMID- 12887872 TI - [Effective psychological interventions for coping with painful medical procedures in pediatric oncology: a theoretical review]. AB - Children with chronic diseases have to undergo numerous and repeated painful medical procedures. Psychological interventions have produced good results in the treatment of this kind of pediatric pain and, although they have not been routinely incorporated into pediatric practice, they provide an effective complement to physical and pharmacological therapies. The present article reviews research into cognitive-behavioral treatment of the distress, pain and anxiety associated with medical procedures in pediatric oncology. We present the possible benefits of these interventions and suggest uses for cognitive-behavioral techniques when performing painful medical procedures. PMID- 12887871 TI - [Appropriateness of treatment of acute pharyngotonsillitis according to the scientific evidence]. AB - OBJECTIVE: To evaluate the appropriateness of antibiotic prescriptions in children with acute pharyngotonsillitis. METHODS: A descriptive study was performed in a series of pediatric patients diagnosed with acute pharyngotonsillitis in the emergency rooms of 11 Spanish hospitals. The appropriateness of antibiotic prescriptions was assessed by comparing our clinical practice in the use of antibiotics for pharyngotonsillitis with consensus guidelines developed for this study. RESULTS: We collected data from 1716 patients with acute pharyngotonsillitis. Antibiotics were prescribed in 80.9 %, mainly according to empirical criteria. The most commonly used antibiotics were amoxicillin (36 %), amoxicillin-clavulanate (22.5 %), cefixime (6.6 %), azithromycin (5.8 %) and cefuroxime (5.2 %). A total of 39.5 % of the patients were aged less than 3 years, of which 75.9 % were treated empirically. Of the prescribed treatments, 22.8 % were considered as the treatment of choice; 22.4 % as alternatives and 54.8 % as inappropriate. CONCLUSIONS: Antibiotic treatment was prescribed in most of the cases of pharyngotonsillitis and nearly always according to empirical criteria. The number of antibiotic prescriptions was far higher than the expected cases of bacterial pharyngotonsillitis and, in many cases, the antibiotic prescriptions were inappropriate. PMID- 12887873 TI - [Ponderal index to describe a term neonatal population]. AB - INTRODUCTION: The terms small, appropriate, and large for gestational age cannot identify some alterations in body composition that might be identified by the ponderal index. The aim of the present study was to correlate birth weight with ponderal index to identify other patterns of intrauterine growth. POPULATION AND METHODS: We performed a cross-sectional, descriptive study in which only one curve of the ponderal index was created for each week of gestational age (weeks 33 to 42) and including the 10th, 50th and 90th centiles because sex does not influence this index. This curve was based on 26,770 healthy infants born in Uruguay and registered in the Perinatal Information System of the Latin American Center for Perinatology. The database consisted of 194,908 infants born between 1995 and 1999. Exclusion criteria were mothers aged less than 15 years or more than 35 years, primiparas, those with > 4 pregnancies, smokers, a diagnosis of intrauterine growth retardation, chronic and pregnancy-related hypertension, cardiac problems, anemic and diabetic mothers, twins with congenital malformations, and intervals between pregnancies of less than 2 years or more than 5 years. Finally, the ponderal index of 43,189 neonates with a gestational age of 40 weeks was compared with the centiles of weight for gestational age to establish alterations in intrauterine growth. RESULTS: Relating the ponderal index with birth weight revealed altered patterns of intrauterine growth in 13 %, 10.3 % and 7.9 % of neonates with appropriate, small and large birth weights, respectively. Moreover, six unusual patters were identified. CONCLUSIONS: Use of the ponderal index curve for gestational age together with other growth curves improves the nutritional assessment of newborns. Further studies should be designed to develop strategies for the immediate, medium- and long-term management of identified risk groups. PMID- 12887874 TI - [Neonatal visits to a pediatric emergency service]. AB - OBJECTIVE: To determine the profile of neonatal visits to a pediatric emergency service and to compare this profile with that of other pediatric age groups. METHOD: We retrospectively reviewed the reports of all neonates who presented to the pediatric emergency service in 2000. Patients transferred from other hospitals were excluded. Age, sex, time of presentation, source of referral, presenting complaint, investigations, final diagnosis and hospitalization were analyzed. RESULTS: Three hundred and nine neonatal visits were identified. The mean age was 14.3 days and 57.3 % were male. Demand was greatest during evening and night shifts and on Sundays. The most common presenting complaints were irritability/crying (19.1 %), constipation (11.7 %) and jaundice (8.7 %). The most frequent diagnoses were infantile colic (16.8 %), constipation (9.7 %) and jaundice (8.7 %). No morbid processes were found in 12.0 % of the patients and complementary investigations were not required in 68.3 %. Fifty-one neonates (16.5 %) were admitted, mainly due to jaundice (9 patients) and sepsis (8 patients). Patients referred by physicians (29 patients, 9.4 %), especially those referred by pediatricians, were admitted and required investigations more often than self-referred patients. The admission rate was higher in neonates than in other pediatric age groups. CONCLUSIONS: Most neonatal utilization of emergency services is due to trivial problems that could be solved in primary care. Appropriate training is required to avoid unnecessary tests without overlooking potentially serious conditions. PMID- 12887876 TI - [Control and assist-control modes of mechanical ventilation]. AB - Control ventilation is a mode of ventilation in which the respirator delivers the preset volume or pressure regardless of the patient's own inspiratory efforts. It is indicated in patients with severe neurological alterations, deep sedation, shock or severe respiratory failure. In this mode, breathing is performed by the respirator, diminishing energy expenditure and reducing the risk of hypo- and hyperventilation. In the assist-control ventilation mode the respirator delivers the preset number of breaths and the patient may also obtain further breaths by making a sufficient respiratory effort to open the triggering sensor. All respiration (preset and patient initiated) is performed by the respirator. Assist control ventilation is indicated inpatients without deep sedation who are able to initiate respiration but who cannot maintain spontaneous respiration. PMID- 12887877 TI - [Male with generalized cutaneous-mucosal hyperpigmentation and neurological deterioration]. PMID- 12887878 TI - [Psychological intervention for coping with painful medical procedures in pediatric oncology]. AB - BACKGROUND: Psychological treatments for procedural distress have shown good results in pediatric oncology and several institutions recommend their implementation to reduce the procedural distress, anxiety and pain associated with painful medical procedures. OBJECTIVES: To assess the results of a psychological intervention in the first child participating in the study "Prospective Analysis of a Psychological Program for Coping with Medical Procedures in Pediatric Oncology". METHODS: We performed a single-case study, using pain and anxiety self-reports by the child and an observational scale in a psychological intervention with the following components: Breathing exercises, imagery, reinforcement, and behavioral rehearsal. RESULTS: The psychological intervention decreased procedural distress, anticipatory anxiety and pain. The 3 year-old child was able to give pain and anxiety self-reports using appropiate tools. CONCLUSIONS: The potential benefits of this kind of intervention are discussed, and some recommendations for future research are proposed. PMID- 12887880 TI - [Scalp myiasis in an immigrant child]. AB - Myiasis is caused by infestation of the skin by the larvae of flies. This entity has specific epidemiological characteristics. Although it is highly unusual in our environment, imported cases can be found because of increased traveling and immigration. The usual clinical manifestations are subcutaneous abscesses, where the larvae develop. Differential diagnosis includes common disorders such as cellulitis and furunculosis, but knowledge of this entity makes diagnosis and treatment simple. We describe the life cycle of the human botfly and the typical clinical presentation and management of this entity. We also highlight the increasing importance of greater awareness of imported diseases. PMID- 12887879 TI - [Sildenafil in the treatment of pulmonary hypertension]. AB - Pulmonary hypertension (PHT) is a rare entity that is difficult to treat. Prognosis is poor. Sildenafil, a selective inhibitor of type 5 phosphodiesterase, has been proposed among the many treatments available for primary and secondary pulmonary hypertension. We report our experience with an infant with pulmonary hypertension due to congenital mitral stenosis and persistent ductus arteriosus, who developed congestive cardiac failure with persistent PHT despite surgical correction. Conventional treatment was unsuccessful and the patient was treated with sildenafil. The clinical course was satisfactory, allowing extubation and withdrawal of vasoactive drugs; pulmonary and left atrial pressure decreased and the patient was discharged. She is currently being treated on an outpatient basis with oral sildenafil and shows satisfactory hemodynamic status. We review alternatives to conventional treatments for pulmonary hypertension with special reference to pediatrics. PMID- 12887881 TI - [Vegetarian diet in glutaric aciduria type I]. AB - Glutaric aciduria type I is an autosomal recessive metabolic disease (1 case/30,000) characterized by a progressive dystonic-diakinetic syndrome in children. Pathologic examination reveals striatal degeneration of the caudate and putamen nucleus and biochemical analysis shows glutaryl CoA dehydrogenase deficiency. Values of glutaric and -hydroxyglutaric acids in urine are usually increased. Currently, the disease is considered untreatable since there are usually irreversible lesions in the central nervous system at diagnosis. However, treatment can be provided to pre-symptomatic children and usually to the siblings of patients with this diagnosis. We present the case of a 23-month-old boy, with macrocephaly and minimal neurologic manifestations at diagnosis, which were attributed to his semivegetarian diet. A dietary regimen and vitamin supplementation halted and even improved symptomatic progression of the disease. We conclude that amino and organic acids in urine should be investigated in all children with progressive macrocephaly of unknown etiology to rule out glutaric aciduria type I. PMID- 12887882 TI - [Hyperpyrexia as a guiding sign for the early diagnosis of x-linked hypohidrotic ectodermic dysplasia]. PMID- 12887883 TI - [Acquired progressive lymphangioma]. PMID- 12887884 TI - [Impact factor and translation to English: apropos of hyperphosphatasemia [corrected]]. PMID- 12887886 TI - Inactivating chromosomes: a macro domain that minimizes transcription. AB - The histone variant macroH2A plays a role in mammalian chromosome X inactivation. Recent data suggest its unusual C-terminal macro domain may be an enzyme that regulates the ADP-ribosylation of chromatin. MacroH2A could thus help correct gene dosage between males and females using a novel epigenetic mark. PMID- 12887887 TI - Cracking the histone code: one, two, three methyls, you're out! AB - In this issue of Molecular Cell, Zhang et al. report the structure of a ternary complex between the SET domain histone methyltransferase DIM-5, its cofactor, and a histone H3 peptide. The insight gained from analysis of a key amino acid provides an exciting opportunity to dissect the possible functional meaning of mono-, di-, and trimethylation of histone lysine residues in vivo that will complement existing approaches in the quest to crack the histone methylation code. PMID- 12887888 TI - Pre-mRNA splicing: awash in a sea of proteins. AB - What's in a spliceosome? More than we ever imagined, according to recent reports employing proteomics techniques to analyze this multi-megadalton machine. As of 1999, around 100 splicing factors were identified (Burge et al., 1999); however, that number has now nearly doubled due primarily to improved purification of spliceosomes coupled with advances in mass spectrometry analyses of complex mixtures. Gratifyingly, most of the previously identified splicing factors were found in the recent mass spec studies. Nonetheless, the number of new proteins emerging with no prior connection to splicing was surprising. Without functional validation, it would be premature to label these proteins as bona fide splicing factors. Yet many were identified multiple times in complexes purified under diverse conditions or from different organisms. Another recurring theme regards the dynamic nature of spliceosomal complexes, which may be even more intricate than previously thought. PMID- 12887889 TI - p53- and Mdm2-independent repression of NF-kappa B transactivation by the ARF tumor suppressor. AB - One mechanism by which a cell affords protection from the transforming effects of oncogenes is via the action of the tumor suppressor, ARF, which activates p53 by inactivating Mdm2. Many oncogenes have also been shown to activate the transcription factor NF-kappa B, which can contribute toward the malignant phenotype in many ways, including an ability to antagonize p53. Here we find that ARF inhibits NF-kappa B function and its antiapoptotic activity independent of Mdm2 and p53. ARF represses the transcriptional activation domain of the NF-kappa B family member RelA by inducing its association with the histone deacetylase, HDAC1. Further, we show that the response of NF-kappa B to the oncogene Bcr-Abl is determined by the ARF status of the cell. These results reveal an important function of ARF that can regulate the NF-kappa B response to oncogene activation. PMID- 12887890 TI - Autoinhibition of Bcr-Abl through its SH3 domain. AB - Bcr-Abl is a dysregulated tyrosine kinase whose mechanism of activation is unclear. Here, we demonstrate that, like c-Abl, Bcr-Abl is negatively regulated through its SH3 domain. Kinase activity, transformation, and leukemogenesis by Bcr-Abl are greatly impaired by mutations of the Bcr coiled-coil domain that disrupt oligomerization, but restored by an SH3 point mutation that blocks ligand binding or a complementary mutation at the intramolecular SH3 binding site defined in c-Abl. Phosphorylation of tyrosines in the activation loop of the catalytic domain and the linker between the SH2 and catalytic domains (SH2-CD linker) is dependent on oligomerization and required for leukemogenesis. These results suggest that Bcr-Abl has a monomeric, unphosphorylated state with the SH3 domain engaged intramolecularly to Pro1124 in the SH2-CD linker, the form that is sensitive to the inhibitor imatinib (STI-571). The sole function of the coiled coil domain is to disrupt the autoinhibited conformation through oligomerization and intermolecular autophosphorylation. PMID- 12887891 TI - PB1 domain-mediated heterodimerization in NADPH oxidase and signaling complexes of atypical protein kinase C with Par6 and p62. AB - Maximal activation of NADPH oxidase requires formation of a complex between the p40(phox) and p67(phox) subunits via association of their PB1 domains. We have determined the crystal structure of the p40(phox)/p67(phox) PB1 heterodimer, which reveals that both domains have a beta grasp topology and that they bind in a front-to-back arrangement through conserved electrostatic interactions between an acidic OPCA motif on p40(phox) and basic residues in p67(phox). The structure enabled us to identify residues critical for heterodimerization among other members of the PB1 domain family, including the atypical protein kinase C zeta (PKC zeta) and its partners Par6 and p62 (ZIP, sequestosome). Both Par6 and p62 use their basic "back" to interact with the OPCA motif on the "front" of the PKC zeta. Besides heterodimeric interactions, some PB1 domains, like the p62 PB1, can make homotypic front-to-back arrays. PMID- 12887892 TI - Sir2 regulates skeletal muscle differentiation as a potential sensor of the redox state. AB - Sir2 is a NAD(+)-dependent histone deacetylase that controls gene silencing, cell cycle, DNA damage repair, and life span. Prompted by the observation that the [NAD(+)]/[NADH] ratio is subjected to dynamic fluctuations in skeletal muscle, we have tested whether Sir2 regulates muscle gene expression and differentiation. Sir2 forms a complex with the acetyltransferase PCAF and MyoD and, when overexpressed, retards muscle differentiation. Conversely, cells with decreased Sir2 differentiate prematurely. To inhibit myogenesis, Sir2 requires its NAD(+) dependent deacetylase activity. The [NAD(+)]/[NADH] ratio decreases as muscle cells differentiate, while an increased [NAD(+)]/[NADH] ratio inhibits muscle gene expression. Cells with reduced Sir2 levels are less sensitive to the inhibition imposed by an elevated [NAD(+)]/[NADH] ratio. These results indicate that Sir2 regulates muscle gene expression and differentiation by possibly functioning as a redox sensor. In response to exercise, food intake, and starvation, Sir2 may sense modifications of the redox state and promptly modulate gene expression. PMID- 12887893 TI - Dynamic interplay of the SUMO and ERK pathways in regulating Elk-1 transcriptional activity. AB - The ETS domain transcription factor Elk-1 is a direct target of the MAP kinase pathways. Phosphorylation of the Elk-1 transcriptional activation domain by MAP kinases triggers its activation. However, Elk-1 also contains two domains with repressive activities. One of these, the R motif, appears to function by suppressing the activity of the activation domain. Here, we demonstrate that SUMO modification of the R motif is required for this repressive activity. A dynamic interplay exists between the activating ERK MAP kinase pathway and the repressive SUMO pathway. ERK pathway activation leads to both phosphorylation of Elk-1 and loss of SUMO conjugation and, hence, to the loss of the repressive activity of the R motif. Thus, the reciprocal regulation of the activation and repressive activities are coupled by MAP kinase modification of Elk-1. PMID- 12887894 TI - Structural basis of degradation signal recognition by SspB, a specificity enhancing factor for the ClpXP proteolytic machine. AB - In prokaryotes, incomplete or misfolded polypeptides emanating from a stalled ribosome are marked for degradation by the addition of an 11 residue peptide (AANDENYALAA) to their C terminus. Substrates containing this conserved degradation signal, the SsrA tag, are targeted to specific proteases including ClpXP and ClpAP. SspB was originally characterized as a stringent starvation protein and has been found to bind specifically to SsrA-tagged proteins and to enhance recognition of these proteins by the ClpXP degradation machine. Here, we report the crystal structures of SspB alone and in complex with an SsrA peptide. Unexpectedly, SspB exhibits a fold found in Sm-family RNA binding proteins. The dimeric SspB structures explain the key determinants for recognition of the SsrA tag and define a hydrophobic channel that may bind unfolded substrates. PMID- 12887895 TI - The CCT chaperonin promotes activation of the anaphase-promoting complex through the generation of functional Cdc20. AB - The WD repeat protein Cdc20 is essential for progression through mitosis because it is required to activate ubiquitin ligation by the anaphase-promoting complex (APC/C). Here we show in yeast that Cdc20 binds to the CCT chaperonin, which is known as a folding machine for actin and tubulin. The CCT is required for Cdc20's ability to bind and activate the APC/C. In vivo, CCT is essential for Cdc20 dependent cell cycle events such as sister chromatid separation and exit from mitosis. The chaperonin is also required for the function of the Cdc20-related protein Cdh1, which activates the APC/C during G1. We propose that folding of the Cdc20 family of APC/C activators is an essential and evolutionary conserved function of the CCT chaperonin. PMID- 12887896 TI - Oligosaccharyltransferase isoforms that contain different catalytic STT3 subunits have distinct enzymatic properties. AB - Oligosaccharyltransferase (OST) is an integral membrane protein that catalyzes N linked glycosylation of nascent proteins in the lumen of the endoplasmic reticulum. Although the yeast OST is an octamer assembled from nonhomologous subunits (Ost1p, Ost2p, Ost3p/Ost6p, Ost4p, Ost5p, Wbp1p, Swp1p, and Stt3p), the composition of the vertebrate OST was less well defined. The roles of specific OST subunits remained enigmatic. Here we show that genomes of most multicellular eukaryotes encode two homologs of Stt3p and mammals express two homologs of Ost3p. The Stt3p and Ost3p homologs are assembled together with the previously described mammalian OST subunits (ribophorins I and II, OST48, and DAD1) into complexes that differ significantly in enzymatic activity. Tissue and cell type specific differences in expression of the Stt3p homologs suggest that the enzymatic properties of oligosaccharyltransferase are regulated in eukaryotes to respond to alterations in glycoprotein flux through the secretory pathway and may contribute to tissue-specific glycan heterogeneity. PMID- 12887897 TI - Splicing-dependent and -independent modes of assembly for intron-encoded box C/D snoRNPs in mammalian cells. AB - In mammalian cells, all small nucleolar RNAs (snoRNAs) that guide rRNA modification are encoded within the introns of host genes. An optimal position about 70 nts upstream of the 3' splice site of the host intron is critical for efficient expression of box C/D snoRNAs in vivo, suggesting synergy with splicing. Here, we have used a coupled in vitro splicing-snoRNA processing system to demonstrate that assembly of box C/D snoRNP proteins is the step affected by snoRNA location, and that active splicing is essential for snoRNP assembly. Splicing blockage experiments further reveal that snoRNP proteins bind specifically at the spliceosomal C1 complex stage. In contrast, splicing independent snoRNP assembly can occur in vitro on snoRNAs that possess stable external stems. In vivo analyses confirm that a stable stem can compensate for the unusual position of those few box C/D snoRNAs located far from the 3' splice site of their host intron. PMID- 12887898 TI - Control of rRNA expression by small molecules is dynamic and nonredundant. AB - The control of ribosomal RNA transcription is one of the most enduring issues in molecular microbiology, having been subjected to intense scrutiny for over 50 years. Rapid changes in rRNA expression occur during transitions in the bacterial growth cycle and following nutritional shifts during exponential growth. Genetic approaches and measurements of initiating nucleoside triphosphate (iNTP) and guanosine 5'-diphosphate, 3'-diphosphate (ppGpp) concentrations and of rRNA promoter activities showed that rapid changes in the concentrations of iNTPs and ppGpp account for the rapid changes in rRNA expression. The two regulatory signals are nonredundant: changes in iNTP concentration dominate regulation during outgrowth from stationary phase, whereas changes in ppGpp concentration are responsible for regulation following upshifts and downshifts during exponential phase. The results suggest a molecular logic for the use of two homeostatic regulatory mechanisms to monitor different aspects of ribosome activity and provide general insights into the nature of overlapping regulatory circuits. PMID- 12887899 TI - Silencing in yeast rDNA chromatin: reciprocal relationship in gene expression between RNA polymerase I and II. AB - About half of approximately 150 rRNA genes are transcriptionally active in Saccharomyces cerevisiae. Chromatin structures in the inactive, and not the active, copies were previously thought to silence both rRNA genes and reporter Pol II genes. Contrary to this belief, we found that silencing of reporters is much stronger in a mutant with approximately 25 rDNA copies, all of which are transcriptionally active. By integrating reporter gene mURA3 with an inactive rDNA copy missing the Pol I promoter, we found that mURA3 is not silenced in chromosomal rDNA repeats. Together with the demonstration of a requirement for active Pol I in silencing, these results show a reciprocal relationship in gene expression between Pol I and Pol II in rDNA. PMID- 12887900 TI - Involvement of actin-related proteins in ATP-dependent chromatin remodeling. AB - Actin-related proteins (Arps) and conventional actin are enigmatic components of many chromatin-remodeling enzyme complexes. The yeast INO80 ATP-dependent chromatin-remodeling complex contains stoichiometric amounts of Arp4, Arp5, Arp8, and actin. Here we have revealed functions of Arp5 and Arp8 by analysis of mutants. arp5 Delta and arp8 Delta mutants display an ino80 Delta phenotype. Purification of INO80 complexes from arp5 Delta and arp8 Delta cells shows that protein complexes remain intact but are compromised for INO80 ATPase activity, DNA binding, and nucleosome mobilization. The INO80 (arp8 Delta) complex is strikingly deficient, not only for the Arp8 subunit, but also for Arp4 and actin, suggesting an ordered assembly of Arps. Binding of Arp8 to the INO80 complex requires an N-terminal region of Ino80 adjacent to the conserved ATPase domain. GST-Arp8 binds preferentially to histones H3 and H4 in vitro, suggesting a histone chaperone function. These findings show direct involvement of Arps in the chromatin-remodeling process. PMID- 12887901 TI - DNA binding and ToxR responsiveness by the wing domain of TcpP, an activator of virulence gene expression in Vibrio cholerae. AB - Virulence in Vibrio cholerae requires activation of toxT by two membrane localized activators, TcpP and ToxR. We isolated 12 tcpP activation mutants that fell into two classes: class I mutants were inactive irrespective of the presence of ToxR, and class II mutants exhibited near wild-type activity when coexpressed with ToxR. Most class I mutants had lesions in the wing domain predicted by homology with the winged helix-turn-helix family of activators. Class I mutants bound promoter DNA poorly and were largely unable to interact with ToxR in a crosslinking assay, whereas class II mutants retained physical interaction with ToxR. One mutant constructed in vitro bound DNA poorly but nevertheless responded to ToxR by activating toxT and also maintained ToxR interaction. We propose that ToxR interaction, but not DNA binding, is essential for TcpP function and that the wing domain of TcpP enables contact with ToxR required for productive TcpP RNA polymerase association. PMID- 12887902 TI - Acetylation of Tat defines a cyclinT1-independent step in HIV transactivation. AB - The HIV transcriptional activator Tat is acetylated by p300 at a single lysine residue in the TAR RNA binding domain. We have generated monoclonal and polyclonal antibodies specific for the acetylated form of Tat (AcTat). Microinjection of anti-AcTat antibodies inhibited Tat-mediated transactivation in cells. Similarly, the p300 inhibitor Lys-CoA and siRNA specific for p300 suppressed Tat transcriptional activity. Full-length synthetic AcTat bound to TAR RNA with the same affinity as unacetylated Tat, but formation of a Tat-TAR CyclinT1 ternary complex was completely inhibited in the presence of AcTat. We propose that Tat acetylation may help in dissociating the Tat cofactor CyclinT1 from TAR RNA and serve to transfer Tat onto the elongating RNA polymerase II. PMID- 12887904 TI - A conformational switch controls the DNA cleavage activity of lambda integrase. AB - The bacteriophage lambda integrase protein (lambda Int) belongs to a family of tyrosine recombinases that catalyze DNA rearrangements. We have determined a crystal structure of lambda Int complexed with a cleaved DNA substrate through a covalent phosphotyrosine bond. In comparison to an earlier unliganded structure, we observe a drastic conformational change in DNA-bound lambda Int that brings Tyr342 into the active site for cleavage of the DNA in cis. A flexible linker connects the central and the catalytic domains, allowing the protein to encircle the DNA. Binding specificity is achieved through direct interactions with the DNA and indirect readout of the flexibility of the att site. The conformational switch that activates lambda Int for DNA cleavage exposes the C-terminal 8 residues for interactions with a neighboring Int molecule. The protein interactions mediated by lambda Int's C-terminal tail offer a mechanism for the allosteric control of cleavage activity in higher order lambda Int complexes. PMID- 12887903 TI - Structural basis for the product specificity of histone lysine methyltransferases. AB - DIM-5 is a SUV39-type histone H3 Lys9 methyltransferase that is essential for DNA methylation in N. crassa. We report the structure of a ternary complex including DIM-5, S-adenosyl-L-homocysteine, and a substrate H3 peptide. The histone tail inserts as a parallel strand between two DIM-5 strands, completing a hybrid sheet. Three post-SET cysteines coordinate a zinc atom together with Cys242 from the SET signature motif (NHXCXPN) near the active site. Consequently, a narrow channel is formed to accommodate the target Lys9 side chain. The sulfur atom of S adenosyl-L-homocysteine, where the transferable methyl group is to be attached in S-adenosyl-L-methionine, lies at the opposite end of the channel, approximately 4 A away from the target Lys9 nitrogen. Structural comparison of the active sites of DIM-5, an H3 Lys9 trimethyltransferase, and SET7/9, an H3 Lys4 monomethyltransferase, allowed us to design substitutions in both enzymes that profoundly alter their product specificities without affecting their catalytic activities. PMID- 12887905 TI - Guarding the genome: electrostatic repulsion of water by DNA suppresses a potent nuclease activity of topoisomerase IB. AB - Type IB topoisomerases cleave and rejoin DNA strands through a stable covalent DNA-(3'-phosphotyrosyl)-enzyme intermediate. The stability of the intermediate is a two-edged sword; it preserves genome integrity during supercoil relaxation, but it also reinforces the toxicity of drugs and lesions that interfere with the DNA rejoining step. Here, we identify a key determinant of the stability of the complex by showing that introduction of an Sp or Rp methylphosphonate linkage at the cleavage site transforms topoisomerase IB into a potent endonuclease. The nuclease reaction entails formation and surprisingly rapid hydrolysis of a covalent enzyme-DNA methylphosphonate intermediate. The approximately 30,000-fold acceleration in the rate of hydrolysis of a methylphosphonate versus phosphodiester suggests that repulsion of water by the DNA phosphate anion suppresses the latent nuclease function of topoisomerase IB. These findings expose an Achilles' heel of topoisomerases as guardians of the genome, and they have broad implications for understanding enzymatic phosphoryl transfer. PMID- 12887906 TI - In vivo roles of Rad52, Rad54, and Rad55 proteins in Rad51-mediated recombination. AB - Repairing a double-strand break by homologous recombination requires binding of the strand exchange protein Rad51p to ssDNA, followed by synapsis with a homologous donor. Here we used chromatin immunoprecipitation to monitor the in vivo association of Saccharomyces cerevisiae Rad51p with both the cleaved MATa locus and the HML alpha donor. Localization of Rad51p to MAT precedes its association with HML, providing evidence of the time needed for the Rad51 filament to search the genome for a homologous sequence. Rad51p binding to ssDNA requires Rad52p. The absence of Rad55p delays Rad51p binding to ssDNA and prevents strand invasion and localization of Rad51p to HML alpha. Lack of Rad54p does not significantly impair Rad51p recruitment to MAT or its initial association with HML alpha; however, Rad54p is required at or before the initiation of DNA synthesis after synapsis has occurred at the 3' end of the invading strand. PMID- 12887907 TI - Recruitment of the recombinational repair machinery to a DNA double-strand break in yeast. AB - Repair of DNA double-strand breaks (DSBs) by homologous recombination requires members of the RAD52 epistasis group. Here we use chromatin immunoprecipitation (ChIP) to examine the temporal order of recruitment of Rad51p, Rad52p, Rad54p, Rad55p, and RPA to a single, induced DSB in yeast. Our results suggest a sequential, interdependent assembly of Rad proteins adjacent to the DSB initiated by binding of Rad51p. ChIP time courses from various mutant strains and additional biochemical studies suggest that Rad52p, Rad55p, and Rad54p each help promote the formation and/or stabilization of the Rad51p nucleoprotein filament. We also find that all four Rad proteins associate with homologous donor sequences during strand invasion. These studies provide a near comprehensive view of the molecular events required for the in vivo assembly of a functional Rad51p presynaptic filament. PMID- 12887908 TI - The coordinated functions of the E. coli MutS and MutL proteins in mismatch repair. AB - The Escherichia coli MutS and MutL proteins have been conserved throughout evolution, although their combined functions in mismatch repair (MMR) are poorly understood. We have used biochemical and genetic studies to ascertain a physiologically relevant mechanism for MMR. The MutS protein functions as a regional lesion sensor. ADP-bound MutS specifically recognizes a mismatch. Repetitive rounds of mismatch-provoked ADP-->ATP exchange results in the loading of multiple MutS hydrolysis-independent sliding clamps onto the adjoining duplex DNA. MutL can only associate with ATP-bound MutS sliding clamps. Interaction of the MutS-MutL sliding clamp complex with MutH triggers ATP binding by MutL that enhances the endonuclease activity of MutH. Additionally, MutL promotes ATP binding-independent turnover of idle MutS sliding clamps. These results support a model of MMR that relies on two dynamic and redundant ATP-regulated molecular switches. PMID- 12887909 TI - BRCA1-independent ubiquitination of FANCD2. AB - Monoubiquitination of the FANCD2 protein is a key step in the Fanconi anemia (FA) tumor suppressor pathway, coinciding with this molecule's accumulation at sites of genome damage. Strong circumstantial evidence points to a requirement for the BRCA1 gene product in this step. Here, we show that the purified BRCA1/BARD1 complex, together with E1 and UbcH5a, is sufficient to reconstitute the monoubiquitination of FANCD2 in vitro. Although siRNA-mediated knockdown of BRCA1 in human cells results in defective targeting of FANCD2 to sites of DNA damage, it does not lead to a defect in FANCD2 ubiquitination. Furthermore, ablation of the RING finger domains of either BRCA1 or BARD1 in the chicken B cell line DT40 also leaves FANCD2 modification intact. Consequently, while BRCA1 affects the accumulation of FANCD2 at sites of DNA damage, BRCA1/BARD1 E3 ligase activity is not essential for the monoubiquitination of FANCD2. PMID- 12887910 TI - A viral mechanism for remodeling chromatin structure in G0 cells. AB - Small DNA viruses force quiescent cells to reenter the cell cycle in order to replicate their DNA. We report here that the adenovirus E1A protein creates an S phase environment in quiescent cells by overcoming the nucleosomal repression of E2F-targeted genes. These genes are surrounded by Lys-9-methylated H3 histones, and their promoters are occupied by the pRb-related protein p130 and the inhibitory transcription factor E2F4. Kinetic analysis indicates that E1A binds to E2F promoters where it eliminates p130 and E2F4, resulting in the dramatic elimination of H3 Lys-9 methylation. Thereafter, H3 Lys-9 acetylation occurs along with the recruitment of activating E2F family members, and this is followed by the transcriptional activity of E2F-targeted genes. These results indicate that E1A has a role in reconfiguring chromatin structure and that this activity is necessary to overcome the repressive mechanisms that maintain cells in a quiescent state. PMID- 12887911 TI - The Sec61p complex is a dynamic precursor activated channel. AB - Previous studies have shown that the rough endoplasmic reticulum (ER) contains nascent precursor polypeptide gated channels. Circumstantial evidence suggests that these channels are formed by the Sec61p complex. We reconstituted the purified Sec61p complex in a lipid bilayer and characterized its dynamics and regulation. The Sec61p complex is sufficient to form the precursor polypeptide activated channel under co- and posttranslational transport conditions. Activity of the Sec61p channel in both transport modes is induced by direct interaction with precursor protein. The Sec61p complex comprises a highly dynamic pore covering conductances corresponding to channel openings from approximately 6 to 60 A. Its properties are indistinguishable from those we observed with native ER channels, directly demonstrating that these channels are formed by the Sec61p complex. PMID- 12887913 TI - CCR5 mimicry by sulfated human anti-HIV-1 antibodies. AB - HIV-1 envelope glycoprotein interaction with a principal coreceptor, CCR5, requires sulfated tyrosines on the CCR5 amino terminus. In this issue of Cell, Choe et al. identified tyrosine-sulfated, neutralizing antibodies against HIV-1 that apparently mimic CCR5. The results demonstrate for the first time that a natural posttranslational modification of an antibody can contribute substantially to antigen recognition. PMID- 12887914 TI - The TNF receptor 1: a split personality complex. AB - The tumor necrosis factor receptor 1 (TNFR1), a prototypic member of the death receptor family signals both cell survival and apoptosis. In this issue of Cell, report that apoptotic TNFR1 signaling proceeds via the sequential formation of two distinct complexes. Since the first complex can activate survival signals and influence the activity of the second complex, this mechanism provides a checkpoint to control the execution of apoptosis. PMID- 12887915 TI - Gene switching by metabolic enzymes--how did you get on the invitation list? AB - Histone gene expression in mammalian cells is codependent upon the Oct-1 transcription factor and its cognate, OCA-S coactivator complex. Surprisingly, GADPH plays an essential role in the OCA-S complex and confers redox dependence upon the in vitro transcription of histone genes. PMID- 12887916 TI - Two poles and a compass. AB - Rho GTPases control fundamental aspects of neutrophil chemotaxis: establishment of front and back and orientation toward the chemoattractant. Two reports in this issue show that activated Cdc42 at the leading edge helps orient the cell's axis in a signaling complex with G beta gamma, PAK1, and PIX alpha; while Rho, activated via G alpha 13, mediates formation of the uropod, which then interacts by mutual negative feedback with the front to reinforce polarization (Li et al., 2003 [this issue of Cell]; Xu et al., [this issue of Cell]). PMID- 12887917 TI - Loading Rho to terminate transcription. AB - In bacteria, one of the major transcriptional termination mechanisms requires a hexameric RNA/DNA helicase known as Rho. One question that has remained unanswered is how the helicase loads onto a nascent transcript so that it can initiate actions on the transcript to cause termination. Recent structures of Rho bound to nucleic acid by show how the individual RNA-binding domains of the 6 subunits are organized and that the ring is split open. The opening is wide enough to accommodate single-stranded RNA and suggests that this conformation is poised to load onto mRNA. PMID- 12887918 TI - Tyrosine sulfation of human antibodies contributes to recognition of the CCR5 binding region of HIV-1 gp120. AB - Sulfated tyrosines at the amino terminus of the principal HIV-1 coreceptor CCR5 play a critical role in its ability to bind the HIV-1 envelope glycoprotein gp120 and mediate HIV-1 infection. Here, we show that a number of human antibodies directed against gp120 are tyrosine sulfated at their antigen binding sites. Like that of CCR5, antibody association with gp120 is dependent on sulfate moieties, enhanced by CD4, and inhibited by sulfated CCR5-derived peptides. Most of these antibodies preferentially associate with gp120 molecules of CCR5-utilizing (R5) isolates and neutralize primary R5 isolates more efficiently than laboratory adapted isolates. These studies identify a distinct subset of CD4-induced HIV-1 neutralizing antibodies that closely emulate CCR5 and demonstrate that tyrosine sulfation can contribute to the potency and diversity of the human humoral response. PMID- 12887919 TI - TIMP-2 mediated inhibition of angiogenesis: an MMP-independent mechanism. AB - Tissue inhibitors of metalloproteinases (TIMPs) suppress matrix metalloproteinase (MMP) activity critical for extracellular matrix turnover associated with both physiologic and pathologic tissue remodeling. We demonstrate here that TIMP-2 abrogates angiogenic factor-induced endothelial cell proliferation in vitro and angiogenesis in vivo independent of MMP inhibition. These effects require alpha 3 beta 1 integrin-mediated binding of TIMP-2 to endothelial cells. Further, TIMP-2 induces a decrease in total protein tyrosine phosphatase (PTP) activity associated with beta1 integrin subunits as well as dissociation of the phosphatase SHP-1 from beta1. TIMP-2 treatment also results in a concomitant increase in PTP activity associated with tyrosine kinase receptors FGFR-1 and KDR. Our findings establish an unexpected, MMP-independent mechanism for TIMP-2 inhibition of endothelial cell proliferation in vitro and reveal an important component of the antiangiogenic effect of TIMP2 in vivo. PMID- 12887920 TI - Induction of TNF receptor I-mediated apoptosis via two sequential signaling complexes. AB - Apoptosis induced by TNF-receptor I (TNFR1) is thought to proceed via recruitment of the adaptor FADD and caspase-8 to the receptor complex. TNFR1 signaling is also known to activate the transcription factor NF-kappa B and promote survival. The mechanism by which this decision between cell death and survival is arbitrated is not clear. We report that TNFR1-induced apoptosis involves two sequential signaling complexes. The initial plasma membrane bound complex (complex I) consists of TNFR1, the adaptor TRADD, the kinase RIP1, and TRAF2 and rapidly signals activation of NF-kappa B. In a second step, TRADD and RIP1 associate with FADD and caspase-8, forming a cytoplasmic complex (complex II). When NF-kappa B is activated by complex I, complex II harbors the caspase-8 inhibitor FLIP(L) and the cell survives. Thus, TNFR1-mediated-signal transduction includes a checkpoint, resulting in cell death (via complex II) in instances where the initial signal (via complex I, NF-kappa B) fails to be activated. PMID- 12887921 TI - Regulation of vertebrate cellular Mg2+ homeostasis by TRPM7. AB - TRPM7 is a polypeptide with intrinsic ion channel and protein kinase domains whose targeted deletion causes cells to experience growth arrest within 24 hr and eventually die. Here, we show that while TRPM7's kinase domain is not essential for activation of its channel, a functional coupling exists such that structural alterations of the kinase domain alter the sensitivity of channel activation to Mg(2+). Investigation of the relationship between Mg(2+) and the cell biological role of TRPM7 revealed that TRPM7-deficient cells become Mg(2+) deficient, that both the viability and proliferation of TRPM7-deficient cells are rescued by supplementation of extracellular Mg(2+), and that the capacity of heterologously expressed TRPM7 mutants to complement TRPM7 deficiency correlates with their sensitivity to Mg(2+). Overall, our results indicate that TRPM7 has a central role in Mg(2+) homeostasis as a Mg(2+) uptake pathway regulated through a functional coupling between its channel and kinase domains. PMID- 12887922 TI - Divergent signals and cytoskeletal assemblies regulate self-organizing polarity in neutrophils. AB - Like neutrophilic leukocytes, differentiated HL-60 cells respond to chemoattractant by adopting a polarized morphology, with F-actin in a protruding pseudopod at the leading edge and contractile actin-myosin complexes at the back and sides. Experiments with pharmacological inhibitors, toxins, and mutant proteins show that this polarity depends on divergent, opposing "frontness" and "backness" signals generated by different receptor-activated trimeric G proteins. Frontness depends upon Gi-mediated production of 3'-phosphoinositol lipids (PI3Ps), the activated form of Rac, a small GTPase, and F-actin. G12 and G13 trigger backness signals, including activation of a second GTPase (Rho), a Rho dependent kinase, and myosin II. Functional incompatibility causes the two resulting actin assemblies to aggregate into separate domains, making the leading edge more sensitive to attractant than the back. The latter effect explains both the neutrophil's ability to polarize in uniform concentrations of chemoattractant and its response to reversal of an attractant gradient by performing a U-turn. PMID- 12887923 TI - Directional sensing requires G beta gamma-mediated PAK1 and PIX alpha-dependent activation of Cdc42. AB - Efficient chemotaxis requires directional sensing and cell polarization. We describe a signaling mechanism involving G beta gamma, PAK-associated guanine nucleotide exchange factor (PIX alpha), Cdc42, and p21-activated kinase (PAK) 1. This pathway is utilized by chemoattractants to regulate directional sensing and directional migration of myeloid cells. Our results suggest that G beta gamma binds PAK1 and, via PAK-associated PIX alpha, activates Cdc42, which in turn activates PAK1. Thus, in this pathway, PAK1 is not only an effector for Cdc42, but it also functions as a scaffold protein required for Cdc42 activation. This G beta gamma-PAK1/PIX alpha/Cdc42 pathway is essential for the localization of F actin formation to the leading edge, the exclusion of PTEN from the leading edge, directional sensing, and the persistent directional migration of chemotactic leukocytes. Although ligand-induced production of PIP(3) is not required for activation of this pathway, PIP(3) appears to localize the activation of Cdc42 by the pathway. PMID- 12887924 TI - Kinesin superfamily protein 2A (KIF2A) functions in suppression of collateral branch extension. AB - Through interactions with microtubules, the kinesin superfamily of proteins (KIFs) could have multiple roles in neuronal function and development. During neuronal development, postmitotic neurons develop primary axons extending toward targets, while other collateral branches remain short. Although the process of collateral branching is important for correct wiring of the brain, the mechanisms involved are not well understood. In this study, we analyzed kif2a(-/-) mice, whose brains showed multiple phenotypes, including aberrant axonal branching due to overextension of collateral branches. In kif2a(-/-) growth cones, microtubule depolymerizing activity decreased. Moreover, many individual microtubules showed abnormal behavior at the kif2a(-/-) cell edge. Based on these results, we propose that KIF2A regulates microtubule dynamics at the growth cone edge by depolymerizing microtubules and that it plays an important role in the suppression of collateral branch extension. PMID- 12887925 TI - Telomerase maintains telomere structure in normal human cells. AB - In normal human cells, telomeres shorten with successive rounds of cell division, and immortalization correlates with stabilization of telomere length. These observations suggest that human cancer cells achieve immortalization in large part through the illegitimate activation of telomerase expression. Here, we demonstrate that the rate-limiting telomerase catalytic subunit hTERT is expressed in cycling primary presenescent human fibroblasts, previously believed to lack hTERT expression and telomerase activity. Disruption of telomerase activity in normal human cells slows cell proliferation, restricts cell lifespan, and alters the maintenance of the 3' single-stranded telomeric overhang without changing the rate of overall telomere shortening. Together, these observations support the view that telomerase and telomere structure are dynamically regulated in normal human cells and that telomere length alone is unlikely to trigger entry into replicative senescence. PMID- 12887926 TI - S phase activation of the histone H2B promoter by OCA-S, a coactivator complex that contains GAPDH as a key component. AB - We have isolated and functionally characterized a multicomponent Oct-1 coactivator, OCA-S which is essential for S phase-dependent histone H2B transcription. The p38 component of OCA-S binds directly to Oct-1, exhibits potent transactivation potential, is selectively recruited to the H2B promoter in S phase, and is essential for S phase-specific H2B transcription in vivo and in vitro. Surprisingly, p38 represents a nuclear form of glyceraldehyde-3-phosphate dehydrogenase, and binding to Oct-1, as well as OCA-S function, is stimulated by NAD(+) but inhibited by NADH. OCA-S also interacts with NPAT, a cyclin E/cdk2 substrate that is broadly involved in histone gene transcription. These studies thus link the H2B transcriptional machinery to cell cycle regulators, and possibly to cellular metabolic state (redox status), and set the stage for studies of the underlying mechanisms and the basis for coordinated histone gene expression and coupling to DNA replication. PMID- 12887929 TI - Parental practices in late adolescence, a comparison of three countries: Canada, France and Italy. AB - The objective of this study was to examine parental practices, such as affection, control and conflict, in three countries: Canada, France and Italy. The sample was composed of more than 900 late adolescents with an average age of 17 years, from three large cities: Montreal, Paris and Rome. Participants answered a self report questionnaire that assessed five measures: emotional bonds with the father and mother, parental supervision, tolerance towards friends, punitiveness for violation of rules, and conflict frequency. Analyses of the results indicated that the country of origin discriminated for most of the parental practice dimensions. Canadian adolescents considered their parents to be more tolerant and rated them as using less punitive measures when rules were broken. Canadian parents also seemed to adopt comparable norms for boys and girls, which could be interpreted as a form of sexual egalitarianism, whereas Italian and French parents appeared less tolerant towards girls. Italian adolescents reported strong emotional bonds with each parent, and also identified more conflicts in their relationships with parents. French adolescents reported weaker emotional bonds with each parent and less parental supervision. Results were interpreted in the light of studies that have reported an influence of cultural background on both parental practices and parental decisions in each of these countries. PMID- 12887930 TI - Individual and familial characteristics of youths involved in street corner gangs in Singapore. AB - Thirty-six youths involved in street corner gangs in Singapore were compared with 91 age-matched controls on measures of self-esteem, aggression, dysfunctional parenting and parent-adolescent communication. Results revealed that gang youths had lower self-esteem and higher levels of aggression than controls. Of note were the findings that gang youths reported a relative absence of parental abuse and indifference, and experienced less open communication and higher levels of over control with their mothers, but not fathers. These findings diverge from anticipated familial correlates of antisocial activity in youths and were discussed with reference to cultural factors and their implications on prevention and intervention. PMID- 12887931 TI - Gender differences in changes in and stability of conduct problems from early adolescence to early adulthood. AB - The present paper examines whether there are gender differences in changes in the aggregate level and individual stability of conduct problems from early adolescence to early adulthood. We analysed self-reported data collected as part of a longitudinal general population study of Norwegian adolescents (n = 1,234). The analyses showed that involvement in conduct problems for both genders peaked in mid-adolescence. The extent to which this peak reflected changes in the number of participants or differences in the frequency of problems among the participants varied according to gender. In most cases, the individual stability of involvement in conduct problems was similar for girls and boys. However, in the long term, relative involvement was more stable for boys than for girls. Moreover, the proportion of boys who showed a high level of involvement both during adolescence and early adulthood was greater than for girls. PMID- 12887932 TI - The relationship between religiosity and adjustment among African-American, female, urban adolescents. AB - African-American female adolescents living in urban environments are at risk for adverse adjustment outcomes, and thus it is imperative to identify protective factors. Religion has been found to be a significant protective resource against many types of maladaptive adjustment outcomes among adolescent samples. The present study accomplishes the following: (1) Provides a description of religiosity in a sample of African-American female teens; (2) examines religion as a resource for these adolescents by focusing on the association between religiosity and sexual activity, self-esteem, and general psychological functioning. Four-hundred ninety-two African-American females, ages 12-19, completed measures on religiosity, sexual activity, self-esteem, and psychological functioning. Most of the adolescents identified as Christian, reported a belief in God, and attended religious services. Greater overall religiosity was associated with greater self-esteem and better psychological functioning. Adolescents at different levels of self-religiosity, as well as family religiosity, evidenced significantly different self-esteem but not psychological distress or sexual activity. Adolescents with varying levels of church attendance demonstrated differences on all three outcomes. By identifying the ways in which religion may exert a positive impact on African-American female teens, mental health professionals can design interventions that have the potential to help improve the quality of life for these adolescents. PMID- 12887933 TI - Parent-child interaction and children's depression: the relationships between parent-child interaction and children's depressive symptoms in Taiwan. AB - This study examines how perceptions of parenting and family messages are associated with the depressive symptoms of early adolescents (N = 454, boys = 224; girls = 210; age, M = 12, S.D. = 1.01) from East Taiwan. Results show that higher levels of parental care and low levels of parental indifference were associated with lower depression scores. Moreover, perceptions of positive messages regarding children themselves, their world, and their future were found to negatively associate with depression, whereas negative messages had an opposite effect. Basically, girls' depressive symptoms were more related to maternal messages, and boys' were more related to paternal messages. PMID- 12887934 TI - The social distribution of explanations of health and illness among adolescents in Sao Paulo, Brazil. AB - The present paper reports on a study, which investigated explanations of health and illness among adolescents from three socio-economic backgrounds in Sao Paulo, Brazil. Interviews and group discussions were used to elicit definitions of health and illness, explanations of illness causality and prevention and maintenance of health among 31 deprived adolescents with recent experience of living on the streets, 20 adolescents from poor families and 20 adolescents from privileged families. The findings showed that adolescents from different socio economic conditions draw upon similar explanatory models to make sense of health and illness. There were also clear distinctions between the groups. Health and illness were presented as being dependent upon multiple factors such as resistance of the body and socio-economic status among privileged adolescents. In contrast, the explanations offered by deprived and poor adolescents for both health and illness rested on behaviour or exposure to environmental hazards. Moreover, drugs, sexually transmitted diseases and faith in God were prominent features of the accounts from deprived adolescents. The findings suggest a need for health professionals to be sensitive to people's perspectives of health and illness and their link to socio-economic conditions. To emphasize individuals' behaviour in the provision of health-related information among deprived adolescents might be counter productive and reinforce victim blaming. PMID- 12887935 TI - Evaluation of the Balloon Analogue Risk Task (BART) as a predictor of adolescent real-world risk-taking behaviours. AB - The Balloon Analogue Risk Task (BART) was administered to a sample of 26 high school-aged adolescents to test the utility of the BART as a behavioural measure of risk-taking propensity. Data indicate that riskyness on the BART was related to self-reported engagement in real-world risk-taking behaviours. These data replicate and uniquely extend past research to an adolescent sample, suggesting that the BART may be a useful addition to self-report batteries for the assessment of risk-taking behaviours in adolescents. PMID- 12887936 TI - Communication about contraception and knowledge of oral contraceptives amongst Norwegian high school students. AB - Communication about contraception and specific knowledge of oral contraceptives (OCs) were examined in a sample (n = 4,650) of Norwegian high school students. Data were collected through a 44-item questionnaire especially developed for the study. The response rate was 73% and data from a total of 1,714 girls and 1,389 boys were eligible for analyses. More females (83%) than males (54%) discussed contraception at least monthly. Discussions were predominantly held with peers and not adults. Females were far more knowledgeable about OCs than males. Respondents knew more about side effects and the pill's relative efficacy than about risks of cancer and tromboembolism. The most significant predictors of high knowledge scores were gender and OC use (females). Also predictive of high knowledge scores were frequent discussions about contraception with peers. Talks about contraception with health workers were unrelated to knowledge scores. PMID- 12887937 TI - Contagion of suicidal symptoms as a function of assortative relating and shared relationship stress in college roommates. AB - Vulnerable people may become socially contiguous via assortative relating, and thus simultaneously susceptible to the effects of shared life stress. To test this possibility, 138 undergraduates and their roommates completed questionnaires on suicidality and stress. Consistent with an assortative relating process, roommates who chose to room together were more similar on a suicide index than were roommates who were assigned to room together. Stress in the roommate relationship amplified similarity in roommates' suicide levels. Results were consistent with the view that shared stress simultaneously affects the suicidality of people whose contiguity was pre-arranged by an assortative relating process. PMID- 12887938 TI - The Teenage Inventory of Social Skills: reliability and validity of the Spanish translation. PMID- 12887942 TI - Thrombosis, prevalence and new evidence on current perceptions of risk. AB - Venous thromboembolism is a significant but unquantified clinical problem in patients with cancer. The FRONTLINE survey was designed to evaluate clinicians' perceptions of thrombosis risk for cancer patients and to examine current clinical practice with regard to thromboprophylaxis and treatment of venous thromboembolism. Over 3800 responses from surgeons, haematologists and other clinicians were received. Their perception of thrombosis risk for patients varied according to the site of primary tumour. Most surgeons thought that patients with brain tumours were at high risk of thrombosis. Most clinicians treating medical (non-surgical) cancer patients perceived pancreatic tumours to carry a high thrombosis risk in the absence of thromboprophylaxis. PMID- 12887943 TI - Are patients with cancer receiving adequate thromboprophylaxis? Results from FRONTLINE. AB - The FRONTLINE survey was designed in part to evaluate thromboprophylaxis regimens currently practised by clinicians worldwide for both surgical and medical patients with cancer. The survey showed that cancer patients undergoing surgery for their malignancy commonly receive thromboprophylaxis, but medical patients with cancer do not, with the exception of patients with a central venous catheter in place. Low-molecular-weight heparin is, overall, the most commonly used thromboprophylactic agent in cancer patients. Oral anticoagulants, however, are often used to prevent thrombosis in medical patients, and are the preferred agent in the USA. The duration of prophylaxis, when administered, is generally longer in medical patients compared with surgical patients. The perception of thrombosis risk for patients with central venous catheters is particularly high in North America where oral anticoagulants are used most commonly, in contrast to other geographical regions where low-molecular-weight heparin is favoured. PMID- 12887944 TI - Are patients with cancer receiving adequate treatment for thrombosis? Results from FRONTLINE. AB - The current practice of clinicians worldwide in the treatment of venous thromboembolism was assessed via responses to the FRONTLINE survey. Significant regional differences in practice were noted. Use of low-molecular-weight heparin was reported as the most common initial treatment for venous thromboembolism, although, in North America, unfractionated heparin was widely used to treat surgical cancer patients after a thrombotic episode. Oral anticoagulants were favoured for long-term treatment of venous thromboembolism, particularly in Western Europe and North America. Case study reports from the survey revealed that a substantial proportion of patients with cancer had been perceived to be at low risk of thrombosis, had consequently not received thromboprophylaxis and developed VTE. PMID- 12887945 TI - Does prolonged thromboprophylaxis improve outcome in patients undergoing surgery? AB - Patients undergoing major abdominal surgery, particularly for malignancy, are at increased risk of venous thromboembolism. Haemostatic markers of coagulation are raised for several weeks after surgery. A higher dose of low-molecular-weight heparin than normally used for thromboprophylaxis is effective in preventing post surgical VTE in patients with cancer with no compromise on bleeding. Four weeks' of thromboprophylaxis with the LMWH dalteparin is significantly more effective than standard (1 week) thromboprophylaxis in preventing proximal DVT. A meta analysis of studies comparing 4 weeks' with 1 week of thromboprophylaxis showed that prolonged thromboprophylaxis with LMWH following major abdominal surgery for malignancy significantly reduces the risk of late occurring DVT. PMID- 12887946 TI - Can we optimise treatment of thrombosis? AB - The occurrence of thromboembolism in patients with cancer complicates their management. Patients with cancer who have established venous thrombosis are at increased risk of recurrent venous thromboembolism and anticoagulant-associated bleeding compared with non-cancer patients. Low-molecular-weight heparins have largely replaced unfractionated heparin as the initial treatment for acute thrombosis and have the advantage that they can be administered at home. The use of oral anticoagulant for the long-term secondary prevention of recurrent venous thromboembolism can be problematic in the cancer patient due to unpredictable changes in the dose response because of poor nutrition, infection, concomitant medications and impaired hepatic function. A large randomised clinical trial has shown that 6 months of treatment with the low-molecular-weight heparin dalteparin in place of 6 months of oral anticoagulant therapy significantly reduces the risk of recurrent thrombosis (50% reduction in risk; p = 0.0017) in cancer patients without an increase in bleeding. PMID- 12887947 TI - An expanding role for antithrombotic therapy in cancer patients. AB - Thromboembolic disease has a negative impact on outcome in the cancer patient. Venous thromboembolism appears to be a more aggressive disease in patients with cancer who present with both a greater thrombus burden and more pronounced derangements of coagulation parameters with non-cancer patients with thrombosis. Restrospective analyses suggest that treatment of venous thromboembolism in cancer patients with low-molecular-weight heparin is associated with additional benefits in terms of their survival. The mechanism for this effect is unclear but may be attributed to a reduced incidence of recurrent thrombosis and fatal pulmonary embolism in those patients with underlying thrombosis. To investigate and potential of low-molecular-weight heparin to prolong survival in cancer patients without underlying thrombosis, a prospective trial (FAMOUS) has evaluated the effect of treatment with the low-molecular-weight heparin dalteparin for 1 year terms of survival of patients with advanced cancer and no underlying thrombosis. PMID- 12887948 TI - Towards complete and accurate reporting of studies of diagnostic accuracy: the STARD initiative. PMID- 12887949 TI - Towards complete and accurate reporting of studies of diagnostic accuracy: the STARD initiative. AB - AIM: To improve the accuracy and completeness of reporting of studies of diagnostic accuracy in order to allow readers to assess the potential for bias in a study and to evaluate the general isability of its results. METHODS: The standards for reporting of diagnostic accuracy (STARD) steering committee searched the literature to identify publications on the appropriate conduct and reporting of diagnostic studies and extracted potential items into an extensive list. Researchers, editors, and members of professional organisations shortened this list during a 2 day consensus meeting with the goal of developing a checklist and a generic flow diagram for studies of diagnostic accuracy. RESULTS: The search for published guidelines about diagnostic research yielded 33 previously published checklists, from which we extracted a list of 75 potential items. At the consensus meeting, participants shortened the list to a 25-item checklist, by using evidence whenever available. A prototype of a flow diagram provides information about the method of recruitment of patients, the order of test execution and the numbers of patients undergoing the test under evaluation, the reference standard, or both. CONCLUSIONS: Evaluation of research depends on complete and accurate reporting. If medical journals adopt the checklist and the flow diagram, the quality of reporting of studies of diagnostic accuracy should improve to the advantage of clinicians, researchers, reviewers, journals, and the public. PMID- 12887950 TI - Refined computed tomography of the thoracic aorta: the impact of electrocardiographic assistance. AB - There have been a number of advances in helical computed tomography (CT) in recent years, which have had a beneficial impact on the quality of imaging of the thoracic aorta. These advances include sub-second gantry rotation, multislice acquisition, and the use of electrocardiographic (ECG) assistance. We examine these techniques with emphasis on the principles behind ECG assistance and its use to reduce aortic motion artefact. We highlight examples of ECG-assisted multislice CT in a spectrum of pathologies of the thoracic aorta. PMID- 12887951 TI - Imaging findings of paediatric oncology patients presenting with acute neurological symptoms. AB - Paediatric oncology patients are prone to central nervous system (CNS) complications due to multiple factors including disorders of the blood cell counts (which include neutropenia, thrombocytopenia or hyperleukocytosis), immunosuppression, neurotoxicity of the treatment, CNS dysfunction due to failure of other organ systems, disease progression of the primary malignancy or metastases. Imaging plays an important role in the management of paediatric oncology patients presenting with acute neurological symptoms. This pictorial review is from our institutional experience on imaging children who are under the care of the Child Cancer Centre. The review consists of a spectrum of neurological complications in paediatric oncology patients. The complications can be classified as (1) cerebrovascular complications, (2) treatment-elated complications, (3) opportunistic infections and (4) tumoural involvement of the CNS. Computed tomography (CT) is the initial choice of investigation, which is easily available and helps to exclude major intracranial abnormality such as haemorrhage. If the CT is negative, magnetic resonance imaging (MRI) should be performed, which is more sensitive for detection of CNS lesions. PMID- 12887952 TI - The diagnostic contribution of the frontal lumbar spine radiograph in community referred low back pain--a prospective study of 1030 patients. PMID- 12887953 TI - The diagnostic contribution of the frontal lumbar spine radiograph in community referred low back pain--a prospective study of 1030 patients. AB - AIM: The diagnostic contribution of the anteroposterior (AP) view was studied to assess whether this view could be omitted safely, thus reducing the radiation burden received by patients undergoing lumbar spine radiography. MATERIALS AND METHODS: Prospective analysis of 1030 consecutive referrals for lumbar spine radiography from general practice. RESULTS: In the majority of cases (90.5%) the AP view was non-contributory. In 4.2% the diagnosis was strengthened and it was altered in 4.6%. However, in the latter group only 1.3% of the total were considered significant alterations. These were cases of possible, but not definite, pars defects and sacroiliitis. Specific important conditions such as infection, malignancy and benign tumours were not missed on the lateral view alone, in our study population. The radiation burden is reduced by 75% by omitting the AP view. CONCLUSION: A single lateral view is an adequate examination, with the proviso that sacroiliac joint disease is not assessed on this view and some pars defects and facet joint degenerative changes may be overlooked. The radiation protection gains are considerable. A single lateral lumbar view is now our routine practice unless sacroiliitis is a specific clinical concern. PMID- 12887954 TI - Abdominal amyloidosis: spectrum of radiological findings. AB - Amyloidosis is a disease characterized by the deposition of fibrillar protein amyloid of beta-structure in organs or tissues. It is usually classified as either a primary disease or secondary to a co-existent condition, such as rheumatoid arthritis, tuberculosis, or neoplasm (particularly multiple myeloma or renal cell carcinoma). Amyloid protein deposition can be seen in a variety of organs though it occurs with higher frequency in the gastrointestinal tract, kidney, and heart. Amyloidosis can have a wide spectrum of manifestations in nearly every abdominal organ. Some of these, for example, multiple cystic submucosal masses of the stomach, amyloidosis of the gallbladder, and dirty soft tissue infiltration of the subcutaneous fat, have not yet been covered in the radiological literature. The combination of various imaging techniques and the identification of characteristic computed tomography (CT) hepatic features may help in the differentiation of amyloidosis from other infiltrative diseases; however, confirmative diagnosis can usually only be achieved by tissue biopsy. PMID- 12887955 TI - Non-Hodgkin's lymphoma of the nasopharynx: CT and MR imaging. AB - OBJECTIVE: Nasopharyngeal (NP) non-Hodgkin's lymphoma (NHL) is an uncommon tumour. The aim of the study was to describe the appearances on CT and MR imaging, and identify the features which help to distinguish NPNHL from other NP tumours. MATERIALS AND METHODS: The CT (n=8) and MR (n=10) images of 14 patients with NPNHL were reviewed retrospectively. Patients with NPNHL were divided into primary NPNHL, where the primary tumour was in the NP (n=7) and secondary NPNHL where the primary tumour was at another extranodal site in the head and neck (n=7). All NPNHL were assessed for tumour size and distribution, appearance and local tumour invasion, in addition lymphadenopathy was assessed in primary NPNHL. RESULTS: The NPNHL ranged in size from 20-75 mm (mean of 55 mm for primary and 30 mm for secondary NHL) and were homogeneous on CT in eight (100%) and MR in seven (70%) and mildly heterogeneous on MR in three (30%) patients. NPNHL involved all walls of the NP in 10 (71%) (n=1). Primary NPNHL extended superficially in five (71%) to involve the nasal cavity (n=3) and oropharynx (n=2) and lymphadenopathy was present in five (71%) being bilateral and involving multiple nodal sites (n=4) with necrosis (n=2) and matting (n=3). CONCLUSION: NPNHL is a homogeneous tumour that tends to diffusely involve all walls of the nasopharynx and spread in an exophytic fashion to fill the airway, rather than infiltrating into the deep tissues. Deep tumour infiltration, when it occurs, is found in those patients with primary NHL and is usually limited in extent and of small volume. and extended in an exophytic fashion to fill the NP cavity in six (43%). Deep tumour invasion was present in two (14%) both patients with primary NHL, the extent and volume of this tumour invasion was small and involved the prevertebral muscles (n=2), parapharyngeal fat space (n=1) and skull base Primary NHL more commonly spreads superficially to involve the nasal cavity or oropharynx, lymphadenopathy is frequent and extensive. A large tumour that fills the nasopharynx, with no or minimal invasion into deep structures, and a propensity to extend down into the tonsil, rather than up into the skull base, may suggest the diagnosis of NHL over nasopharyngeal carcinoma. PMID- 12887956 TI - The prevalence of simple renal and hepatic cysts detected by spiral computed tomography. AB - AIM: To provide a definitive evaluation of the prevalence of simple renal and hepatic cysts using spiral computed tomography (CT). PATIENTS AND METHODS: Contrast-enhanced abdominal CT studies from 617 patients (295 women, 322 men) attending for investigations unrelated to renal or hepatic pathology were included. The number of renal cysts, their location and the diameter of the largest cyst were recorded. Hepatic cysts were recorded as being either present or absent. RESULTS: Two hundred and fifty-four patients (41%) were found to have simple renal cysts and 110 patients were found to have simple hepatic cysts (18%). Both renal and hepatic cysts became more common with age. Renal cysts were significantly more common at all ages in men (p=0.001), and increased in both size (p=0.02) and number (p<0.05) with age. CONCLUSION: A major discrepancy has, until now, existed between autopsy prevalence of renal cysts and prevalence as reported by in-vivo third-generation CT or sonography studies. Using newer spiral CT machines, we have generated prevalence data closer to post-mortem findings. We have also shown a significant, but age dependent, association between the presence of simple hepatic and simple renal cysts (p=0.001) PMID- 12887957 TI - Histopathological and CT features of pulmonary sclerosing haemangiomas. AB - AIM: To demonstrate the computed tomography (CT) features of pulmonary sclerosing haemangiomas. MATERIALS AND METHODS: Six pathologically proven sclerosing haemangiomas were included in this retrospective review. Patients consisted of five women and one man aged 20-54 years (mean, 34.5 years). Their CT features were recorded according to enhancement patterns and the presence of a tail sign, prominent pulmonary artery sign, air-trapping sign, presence of calcification or cystic spaces, consolidation, and interstitial infiltration. The predominant composition of an individual sclerosing haemangioma was documented by means of microscopy. RESULTS: Inhomogeneous enhancement was frequently present and depended on the various compositions of the tumours, especially those in sclerotic and predominantly papillary predominant types. The presence of a tail sign, intra-tumoural cystic areas, and a prominent artery sign were the frequent features in our cases. CONCLUSION: Sclerosing haemangioma should be considered in young or middle-aged female patients whose CT images show them having an inhomogeneous enhancing soft tissue mass with a smooth outline and with above the aforementioned features. PMID- 12887958 TI - Sonographic appearances of medial retinacular complex injury in transient patellar dislocation. AB - AIM: To describe the sonographic appearances of the medial retinacular (MPFR) complex of the knee in patients with acute and recurrent patellar dislocation. MATERIALS AND METHODS: Thirty patients were scanned within 2-4 weeks of an acute episode of lateral patellar dislocation. Eleven gave a history of recurrent patellar dislocation. Ten patients had examination under anaesthesia with arthroscopy and repair of the injury. The sonographic and operative results were compared. RESULTS: The normal sonographic appearance of the MPFR is described. Of the 10 patients who underwent examination under anaesthesia, four patients had complete avulsion of the MPFR from the patella, two patients had avulsion of the MPFR from the adductor tubercle and four patients had avulsion of the MPFR from both the patella and adductor tubercle. There was complete correlation between the sonographic and operative findings for injuries of the MPFR. Other findings included partial retinacular tears, injury to the medial collateral ligament, haematoma within vastus medialis obliquus (VMO) and bony avulsions from the patella and adductor tubercle CONCLUSION: Sonography gives reliable information regarding the site of the injury and its extent thus helping to decide whether conservative or operative treatment is the most appropriate approach to management of the injury. PMID- 12887959 TI - CT evaluation of nasolacrimal relapse of nasopharyngeal carcinoma. AB - AIM: The aim of the current study was to elucidate the anatomy of the nasolacrimal apparatus using computed tomography (CT), and to investigate the CT findings suggestive of recurrent nasopharyngeal carcinoma (NPC) involving the nasolacrimal apparatus. MATERIALS AND METHODS: CT images of five patients with clinical or radiological findings suggestive of recurrent NPC who presented with nasolacrimal involvement were reviewed. Involvement of the lacrimal fossa, nasolacrimal canal (NLC), and adjacent anatomic structures including orbit and sinonasal cavities were noted. RESULTS: CT in all patients demonstrated findings of NLC invasion and ostiomeatal complex obliteration. Four of the five patients showed medial canthal mass or pre-septal thickening, ethmoid bulla opacification and inferior nasal meatal involvement. CONCLUSION: Knowledge of the anatomy of the nasolacrimal apparatus using CT is critical for evaluating images for tumour recurrence at this site. NLC invasion, a medial canthal mass and pre-septal thickening are important primary indicators for potential involvement of the nasolacrimal apparatus by recurrent NPC. Secondary signs suggesting this distinct form of relapse are obliteration of ostiomeatal channel, ethmoid bulla opacification and inferior nasal meatus involvement. PMID- 12887960 TI - Thunderclap headache: presentation of intracranial sinus thrombosis? PMID- 12887961 TI - Mediastinal schwannoma: MR imaging findings of an unusual case presenting as a lobulated mass with internal fibrous septa. PMID- 12887962 TI - Pregnancy after treatment for Hodgkin's lymphoma: inadvertent diagnosis by CT. PMID- 12887963 TI - Neuroimaging for non-accidental head injury in childhood: a proposed protocol. PMID- 12887969 TI - Signaling between SR and plasmalemma in smooth muscle: sparks and the activation of Ca2+-sensitive ion channels. AB - Intracellular calcium ions are involved in the regulation of nearly every aspect of cell function. In smooth muscle, Ca2+ can be delivered to Ca2+-sensitive effector molecules either by influx through plasma membrane ion channels or by intracellular Ca2+ release events. Ca2+ sparks are transient local increases in intracellular Ca2+ that arise from the opening of ryanodine-sensitive Ca2+ release channels (ryanodine receptors) located in the sarcoplasmic reticulum. In arterial myocytes, Ca2+ sparks occur near the plasma membrane and act to deliver high (microM) local Ca2+ to plasmalemmal Ca2+-sensitive ion channels, without directly altering global cytosolic Ca2+ concentrations. The two major ion channel targets of Ca2+ sparks are Ca2+-activated chloride (Cl(Ca)) channels and large conductance Ca2+-activated potassium (BK) channels. The activation of BK channels by Ca2+ sparks play an important role in the regulation of arterial diameter and appear to be involved in the action of a variety of vasodilators. The coupling of Ca2+ sparks to BK channels can be influenced by a number of factors including membrane potential and modulatory beta subunits of BK channels. Cl(Ca) channels, while not present in all smooth muscle, can also be activated by Ca2+ sparks in some types of smooth muscle. Ca2+ sparks can also influence the activity of Ca2+ dependent transcription factors and expression of immediate early response genes such as c-fos. In summary, Ca2+ sparks are local Ca2+ signaling events that in smooth muscle can act on plasma membrane ion channels to influence excitation contraction coupling as well as gene expression. PMID- 12887970 TI - Sodium-calcium exchange influences the response to endothelin-1 in lens epithelium. AB - Studies were conducted to examine the possible involvement of Na+-Ca2+ exchanger in determining the magnitude of the endothelin-1 (ET-1)-receptor-mediated calcium signal in porcine lens epithelial cells. Cytoplasmic calcium concentration was measured in primary cultured cells loaded with Fura-2. ET-1 (100 nM) caused cytoplasmic calcium to increase transiently to approximately 250 nM from a baseline of approximately 65 nM. The calcium increase decayed to a sustained plateau 35-45 nM above the baseline. Both the peak and plateau component of the ET-1 calcium response were abolished by PD145065, an ET receptor antagonist, and by cyclopiazonic acid (CPA) (10 microM). In calcium-free bathing solution, only the plateau was abolished. In the presence of ouabain, low-sodium bathing solution or bepridil, a sodium-calcium exchange inhibitor, peak height more than doubled. Bepridil also increased the peak height of the calcium response to ATP. The half-time for decay of the ET-1 and ATP calcium peak was increased several folds by bepridil, ouabain and low-sodium conditions. Measurements of ionomycin releasable calcium suggested calcium store size was not increased in bepridil treated cells. Taken together findings suggest inhibition of sodium-calcium exchange increases the magnitude of the receptor-initiated store-release phase of the ET-1 calcium signaling response as the result of impaired calcium clearance from the cytoplasm. PMID- 12887971 TI - Evidence that store-operated Ca2+ channels are more effective than intracellular messenger-activated non-selective cation channels in refilling rat hepatocyte intracellular Ca2+ stores. AB - Liver cells possess store-operated Ca2+ channels (SOCs) with a high selectivity for Ca2+ compared with Na+, and several types of intracellular messenger activated non-selective cation channels with a lower selectivity for Ca2+ (NSCCs). The main role of SOCs is thought to be in refilling depleted endoplasmic reticulum Ca2+ stores [Cell Calcium 7 (1986) 1]. NSCCs may be involved in refilling intracellular stores but are also thought to have other roles in regulating the cytoplasmic-free Ca2+ and Na+ concentrations. The ability of SOCs to refill the endoplasmic reticulum Ca2+ stores in hepatocytes has not previously been compared with that of NSCCs. The aim of the present studies was to compare the ability of SOCs and maitotoxin-activated NSCCs to refill the endoplasmic reticulum in rat hepatocytes. The experiments were performed using fura-2FF and fura-2 to monitor the free Ca2+ concentrations in the endoplasmic reticulum and cytoplasmic space, respectively, a Ca2+ add-back protocol, and 2-aminoethyl diphenylborate (2-APB) to inhibit Ca2+ inflow through SOCs. In cells treated with 2,5-di-t-butylhydroquinone (DBHQ) or vasopressin to deplete the endoplasmic reticulum Ca2+ stores, then washed to remove DBHQ or vasopressin, the addition of Ca2+ caused a substantial increase in the concentration of Ca2+ in the endoplasmic reticulum and cytoplasmic space due to the activation of SOCs. These increases were inhibited 80% by 2-APB, indicating that Ca2+ inflow is predominantly through SOCs. In the presence of 2-APB (to block SOCs), maitotoxin induced a substantial increase in [Ca2+](cyt), but only a modest and slower increase in [Ca2+](er). Under these conditions, Ca2+ inflow is predominantly through maitotoxin-activated NSCCs. It is concluded that SOCs are more effective than maitotoxin-activated NSCCs in refilling the endoplasmic reticulum Ca2+ stores. The previously developed concept of a specific role for SOCs in refilling the endoplasmic reticulum is consistent with the results reported here. PMID- 12887972 TI - Physiological Ca2+ level and Ca2+-induced Permeability Transition Pore control protein phosphorylation in rat brain mitochondria. AB - Phosphorylation of several low molecular mass proteins (3.5, 17, 23 and 29kDa) was observed in rat brain mitochondria (RBM) at ATP concentration close to that in the mitochondrial matrix. Furthermore, regulatory effects of Ca2+ on phosphorylation of these proteins were investigated. Protein phosphorylation was found to be modulated by Ca2+ in the physiological concentration range (10(-8) to 10(-6)M free Ca2+). Incorporation of 32P from [gamma-32P]ATP into the 17kDa protein was dramatically increased within the 10(-7) to 10(-6)M free Ca2+ range, whereas an opposite effect was observed for the 3.5kDa polypeptide. Strong de phosphorylation of the 3.5kDa polypeptide and enhanced 32P-incorporation into the 17 and 23kDa proteins were found with supra-threshold Ca2+ loads and these effects were eliminated or reduced in the presence of cyclosporin A, an inhibitor of Permeability Transition Pore (PTP) opening. In the presence of calmidazolium (Cmz), a calmodulin antagonist, enhanced levels of phosphorylation of the 17 and 3.5kDa polypeptides were observed and the 17kDa protein phosphorylation was suppressed by H-8, a protein kinase A inhibitor. It is concluded that Ca2+ in physiological concentrations, as a second messenger, can control phosphorylation of the low molecular mass phospoproteins in RBM, in addition to well known regulation of some Krebs cycle dehydrogenases by Ca2+. The protein phosphorylation was strongly dependent on the Ca2+-induced PTP opening. PMID- 12887973 TI - Vesl/Homer proteins regulate ryanodine receptor type 2 function and intracellular calcium signaling. AB - Cellular signaling proteins such as metabotropic glutamate receptors, Shank, and different types of ion channels are physically linked by Vesl (VASP/Ena-related gene up-regulated during seizure and LTP)/Homer proteins [Curr. Opin. Neurobiol. 10 (2000) 370; Trends Neurosci. 23 (2000) 80; J. Cell Sci. 113 (2000) 1851]. Vesl/Homer proteins have also been implicated in differentiation and physiological adaptation processes [Nat. Neurosci. 4 (2001) 499; Nature 411 (2001) 962; Biochem. Biophys. Res. Commun. 279 (2000) 348]. Here we provide evidence that a Vesl/Homer subtype, Vesl-1L/Homer-1c (V-1L), reduces the function of the intracellular calcium channel ryanodine receptor type 2 (RyR2). In contrast, Vesl-1S/Homer-1a (V-1S) had no effect on RyR2 function but reversed the effects of V-1L. In live cells, in calcium release studies and in single-channel electrophysiological recordings of RyR2, V-1L reduced RyR2 activity. Important physiological functions and pharmacological properties of RyR2 are preserved in the presence of V-1L. Our findings demonstrate that a protein-protein interaction between V-1L and RyR2 is not only necessary for organizing the structure of intracellular calcium signaling proteins [Curr. Opin. Neurobiol. 10 (2000) 370; Trends Neurosci. 23(2000)80; J. Cell Sci. 113 (2000) 1851; Nat Neurosci. 4 (2001) 499; Nature 411 (2001) 962; Biochem. Biophys. Res. Commun. 279 (2000) 348; Nature 386 (1997) 284], but that V-1L also directly regulates RyR2 channel activity by changing its biophysical properties. Thereby it may control cellular calcium homeostasis. These observations suggest a novel mechanism for the regulation of RyR2 and calcium-dependent cellular functions. PMID- 12887974 TI - Ca2+ transients of cardiomyocytes from senescent mice peak late and decay slowly. AB - Ventricular myocytes were isolated from either young (2 months, "young myocytes") or senescent (20-26 months, "senescent myocytes") mice. Ca2+ transients were evoked by 40ms voltage-clamp pulses depolarising at 0.4, 1, 2, 4 or 8Hz. At 8Hz, Ca2+ transients from senescent cells peaked later (39ms versus 23ms) to smaller systolic [Ca2+](c) (667nM versus 1110nM) and decayed at slower rate (16s(-1) versus 33s(-1)) to higher end-diastolic [Ca2+](c) (411nM versus 220nM) than those from young myocytes. These differences were less pronounced at lower frequencies of pulsing and could not be explained by differences of the time integral of Ca2+ inward current. Since concentrations of SERCA2a and SERCA2b proteins were similar in young and senescent cells, slow rate of Ca2+ decay and high diastolic [Ca2+]c are explained on the assumption that the usual Ca2+ stimulation of SERCA2 activity is attenuated in senescent cells. The prolonged time-to-peak [Ca2+]c is discussed to result from insufficient SR Ca2+ filling by SERCA2 and, in context with confocal images, from a shift of the SERCA2b distribution to the subsarcolemmal space. The age-related changes of the Ca2+ transients are discussed to cause systolic and diastolic failure if senescent mouse hearts beat at high frequencies. PMID- 12887975 TI - A new mitochondrial fluorescent zinc sensor. AB - A novel cationic fluorescent zinc (Zn2+) indicator (RhodZin-3) with nanomolar affinity for Zn2+ has been synthesized. RhodZin-3 exhibits large pH-independent fluorescence increases in the orange region of the visible wavelength spectrum with increasing zinc concentrations, and no sensitivity to physiologically relevant Ca2+ concentrations. Experiments in neuronal cell cultures show that RhodZin-3 effectively localizes into mitochondria and detects changes of intramitochondrial free Zn2+ ([Zn2+]m). PMID- 12887976 TI - Hg2+ signaling in trout hepatoma (RTH-149) cells: involvement of Ca2+-induced Ca2+ release. AB - Mercury is a non-essential heavy metal affecting intracellular Ca2+ dynamics. We studied the effects of Hg2+ on [Ca2+]i in trout hepatoma cells (RTH-149). Confocal imaging of fluo-3-loaded cells showed that Hg2+ induced dose-dependent, sustained [Ca2+]i transient, triggered intracellular Ca2+ waves, stimulated Ca2+ ATPase activity, and promoted InsP3 production. The effect of Hg2+ was reduced by the Ca2+ channel blocker verapamil and totally abolished by extracellular GSH, but was almost unaffected by cell loading with the heavy metal chelator TPEN or esterified GSH. In a Ca2+-free medium, Hg2+ induced a smaller [Ca2+]i transient, that was unaffected by TPEN, but was abolished by U73122, a PLC inhibitor, and by cell loading with GDP-betaS, a G protein inhibitor, or heparin, a blocker of intracellular Ca2+ release. Data indicate that Hg2+ induces Ca2+ entry through verapamil-sensitive channels, and intracellular Ca2+ release via a G protein-PLC InsP3 mechanism. However, in cells loaded with heparin and exposed to Hg2+ in the presence of external Ca2+, the [Ca2+]i rise was maximally reduced, indicating that the global effect of Hg2+ is not a mere sum of Ca2+ entry plus Ca2+ release, but involves an amplification of Ca2+ release operated by Ca2+ entry through a CICR mechanism. PMID- 12887977 TI - Monitoring neuronal calcium signalling using a new method for ratiometric confocal calcium imaging. AB - Ca2+ signalling influences many processes in the adult and developing nervous system like exocytosis, synaptic plasticity, and growth cone motility. Optical techniques in combination with fluorescent Ca2+ indicators are the most frequently used methods to measure Ca2+ signalling in cells. In the present study, a new method for ratiometric confocal Ca2+ imaging was developed, and the usefulness of the system was tested with two different neuronal preparations. Developing Manduca sexta antennal lobe neurons were loaded with the Ca2+ sensitive dye Fura Red-AM, and the ratio of fluorescence excited at 457 and 488nm was measured with a confocal laser scanning microscope. During pupal stages 4-12, the antennal lobe neuropil is restructured which includes the ingrowth of olfactory receptor axons, dendritic outgrowth of antennal lobe neurons, and synaptogenesis. In antennal lobe neurons, application of the AChR agonist carbachol induced Ca2+ oscillations the amplitude and frequency of which changed during stages 4-9, while at the end of synaptogenesis, at stages 11 and 12, only single Ca2+ transients were elicited. The Ca2+ oscillations were blocked by D tubocurarine and Cd2+, indicating that they were due to Ca2+ influx through voltage-gated Ca2+ channels, activated by nAChR-mediated membrane depolarization. To test whether single action potentials can induce Ca2+ transients detectable by Fura Red, individual leech Retzius neurons were injected iontophoretically with the Ca2+ indicator, and the membrane potential was recorded during Ca2+ imaging. Single action potentials induced transient increases in the Fura Red ratio measured in the axon, while trains of action potentials elicited Ca2+ transients that could also be recorded in the cell body and the nucleus. The results show that Fura Red can be used as a ratiometric Ca2+ indicator for confocal imaging. PMID- 12887978 TI - A tale of two tails: cytosolic termini and K(+) channel function. AB - The enormous variety of neuronal action potential waveforms can be ascribed, in large part, to the sculpting of their falling phases by currents through voltage gated potassium channels. These proteins play several additional roles in other tissues such as the regulation of heartbeat and of insulin release from pancreatic cells as well as auditory signal processing in the cochlea. The functional channel is a tetramer with either six or two transmembrane segments per monomer. Selectivity filters, voltage sensors and gating elements have been mapped to residues within the transmembrane region. Cytoplasmic residues, which are accessible targets for signal transduction cascades and provide attractive means of regulation of channel activity, are now seen to be capable of modulating various aspects of channel function. Here we review structural studies on segments of the cytoplasmic tails of K(+) channels, as well as the range of modulatory activities of these tails. PMID- 12887979 TI - Calcium-induced calcium release in smooth muscle: the case for loose coupling. AB - This article reviews the key experiments demonstrating calcium-induced calcium release (CICR) in smooth muscle and contrasts the biophysical and molecular features of coupling between the sarcolemmal (L-type Ca(2+) channel) and sarcoplasmic reticulum (ryanodine receptor) Ca(2+) channels in smooth and cardiac muscle. Loose coupling refers to the coupling process in smooth muscle in which gating of ryanodine receptors is non-obligate and may occur with a variable delay following opening of the sarcolemmal Ca(2+) channels. These features have been observed in the earliest studies of CICR in smooth muscle and are in marked contrast to cardiac CICR, where a close coupling between T-tubular and SR membranes results in tight coupling between the gating events. The relationship between this "loose coupling" and distinct subcellular release sites within smooth muscle cells, termed frequent discharge sites, is discussed. PMID- 12887980 TI - Channels as taste receptors in vertebrates. AB - Taste reception is fundamental for proper selection of food and beverages. Chemicals detected as taste stimuli by vertebrates include a large variety of substances, ranging from inorganic ions (e.g., Na(+), H(+)) to more complex molecules (e.g., sucrose, amino acids, alkaloids). Specialized epithelial cells, called taste receptor cells (TRCs), express specific membrane proteins that function as receptors for taste stimuli. Classical view of the early events in chemical detection was based on the assumption that taste substances bind to membrane receptors in TRCs without permeating the tissue. Although this model is still valid for some chemicals, such as sucrose, it does not hold for small ions, such as Na(+), that actually diffuse inside the taste tissue through ion channels. Electrophysiological, pharmacological, biochemical, and molecular biological studies have provided evidence that indeed TRCs use ion channels to reveal the presence of certain substances in foodstuff. In this review, we focus on the functional and molecular properties of ion channels that serve as receptors in taste transduction. PMID- 12887981 TI - Defective pantomime of object use in left brain damage: apraxia or asymbolia? AB - Disturbance of pantomime of object use in patients with left brain damage (LBD) and aphasia has been firmly established but its nature remains controversial. It may be due to an inability to perform movements from memory without external support by objects (apraxia) or to an inability to produce signs referring to absent objects and actions (asymbolia). The need to perform movements without external support is shared with imitation of gestures, and the demand to designate absent objects with drawing from memory. Both of these tasks have been found to be impaired in LBD. We examined pantomime of object use, drawing objects from memory, imitation of meaningless gestures, and aphasia in 40 patients with LBD and aphasia and compared them to healthy controls and to patients with right brain damage (RBD). Whereas drawing showed comparable sensitivity to LBD and RBD, pantomime was distinctly more disturbed in LBD than in RBD patients. Pantomime was worse than drawing in LBD but better than drawing in RBD. In the LBD group scores on pantomime showed significant correlations of very similar strength to drawing, imitation, and all language tests. Multidimensional scaling of the correlational structure placed pantomime in an intermediate position between verbal and non-verbal tests. We conclude that neither apraxia nor asymbolia can satisfactorily explain our results. It seems as if pantomime of object use taps a central aspect of left hemisphere function which is compromised by any LBD. We propose that this may be the ability to select and combine distinctive features of objects and actions. PMID- 12887982 TI - Abnormalities of brain function during a nonverbal theory of mind task in schizophrenia. AB - Theory of mind (ToM), the specific ability to attribute thoughts and feelings to oneself and others is generally impaired in schizophrenia. Previous studies demonstrated a deficit of the attribution of intentions to others among patients having formal thought disorder. During nonverbal tasks, such a function requires both the visual perception of human figures and the understanding of their intentions. These processes are considered to involve the superior temporal sulcus and the medial prefrontal cortex, respectively. Are the functional patterns of activation associated with those processes abnormal in schizophrenia? Seven schizophrenic patients on medication performed a nonverbal attribution of intentions task as well as two matched physical logic tasks, with and without human figures, while H2O15 PET-scanning was performed. Data from the patients were compared to those of eight healthy controls matched for verbal IQ and sex. The experimental design allowed dissociating the effect of the perception of human figures from that of the attribution of intentions. During attribution of intentions, significant activations in the right prefrontal cortex were detected in the control subjects. Those activations were not found in the schizophrenic group. However, in both groups, the perception of human figure elicited bilateral activation of the occipitotemporal regions and of the posterior part of the superior temporal sulcus. Schizophrenic patients performing a nonverbal attribution of intentions task have an abnormal cerebral activity. PMID- 12887983 TI - Brain activation patterns during measurement of sub- and supra-second intervals. AB - The possibility that different neural systems are used to measure temporal durations at the sub-second and several second ranges has been supported by pharmacological manipulation, psychophysics, and neural network modelling. Here, we add to this literature by using fMRI to isolate differences between the brain networks which measure 0.6 and 3s in a temporal discrimination task with visual discrimination for control. We observe activity in bilateral insula and dorsolateral prefrontal cortex, and in right hemispheric pre-supplementary motor area, frontal pole, and inferior parietal cortex during measurement of both intervals, suggesting that these regions constitute a system used in temporal discrimination at both ranges. The frontal operculum, left cerebellar hemisphere and middle and superior temporal gyri, all show significantly greater activity during measurement of the shorter interval, supporting the hypotheses that the motor system is preferentially involved in the measurement of sub-second intervals, and that auditory imagery is preferentially used during measurement of the same. Only a few voxels, falling in the left posterior cingulate and inferior parietal lobe, are more active in the 3s condition. Overall, this study shows that although many brain regions are used for the measurement of both sub- and supra-second temporal durations, there are also differences in activation patterns, suggesting that distinct components are used for the two durations. PMID- 12887984 TI - Anomia for people names in DAT--evidence for semantic and post-semantic impairments. AB - Proper name anomia is a frequent finding among patients in the early stages of Alzheimer's disease. The present study investigates naming of famous persons in a group of DAT patients, a group of persons with mild cognitive impairment (MCI) and healthy controls. The study is aimed at distinguishing the relative contributions of semantic and post-semantic factors to difficulties in proper name retrieval. As shown by a significantly lower score in answering semantic questions, DAT patients retrieve less biographical knowledge related to famous persons than healthy elderly subjects and persons with mild cognitive impairment. This finding is in line with the frequent observation of semantic deficits in early and moderate DAT. The high number of Tip-of-the-Tongue (TOT) answers in DAT found in relation to few spontaneously named items shows that post-semantic deficits are as important as semantic deficits in determining anomia for people names in DAT. Moreover, DAT patients were less sensitive to phonological cueing than healthy persons or persons with mild cognitive impairment. These findings suggest that proper name anomia in DAT is not only due to semantic deficits, but also to problems in accessing the phonological representation, as well as to a degradation of phonological representations. Thus, naming deficits in DAT differ not only quantitatively, but also qualitatively from the difficulties of healthy elderly persons. No significant differences were found between persons with mild cognitive impairment and healthy controls in proper name retrieval. PMID- 12887985 TI - Relationships between time estimation, memory, attention, and processing speed in patients with severe traumatic brain injury. AB - The present experiment was aimed at investigating the effects of memory and attention deficits and of information processing slowing on time estimation in patients with severe traumatic brain injury (TBI). Patients with TBI and normal control subjects reproduced and produced durations (5, 14, 38s) in both a control counting condition and in a concurrent reading condition. They also performed finger-tapping tasks at a free rate and at a 1s rate. Both groups were assessed on processing speed using a reaction-time task, and on memory and attention using a battery of neuropsychological tests. The results showed that time estimation was not less accurate in patients with TBI than in control subjects on the reproduction task or on the production task performed either in the control counting condition or in the concurrent reading condition. Conversely, duration judgments were more variable in patients with TBI than in control subjects on both tasks in both conditions. The results also showed that TBI patients exhibited slower reaction-times, and poorer working and episodic memory scores than control subjects. Most importantly, the variability index in the duration reproduction task was related to both working memory scores and processing speed measures, whereas the variability index in the duration production task was only related with the processing speed measures. The temporal performance pattern in TBI patients does not appear to reflect specific deficits in timing, but rather overall problems in attention, working memory, and processing speed mechanisms. PMID- 12887986 TI - Transcoding zeros within complex numerals. AB - This paper describes a patient (LD) showing a selective syntactic deficit in the production of Arabic numerals. Unlike in previously reported cases, LD's syntactic difficulties result in deletions rather than insertions of zeros, with a reduction of the number magnitude. The pattern of errors highlighted a distinction between "lexical zeros", i.e. the zeros in tens, that are semantically derived, and "syntactic zeros" that are syntactically produced as the result of specific production rules. In LD, only syntactic zeros were affected. Furthermore, the processing of numerals with final zeros was found to be easier than the processing of numerals with internal zeros. This pattern of errors is compatible with the lexical-semantic model of Power and Dal Martello. In this model, in fact, lexical zeros originate from a numerical concept, while syntactic zeros originate from a concatenation operation, plus an overwriting operation leaving one or more intermediary zeros. Thus, lexical zeros may be easier to manipulate than syntactic zeros that merely represent a null quantity associated to a specific power of 10. PMID- 12887987 TI - Perception of action boundaries in patients with frontal lobe damage. AB - The prefrontal cortex is known to be involved in action planning and in controlling behaviour. Neuropsychological evidence also supports the idea that the prefrontal cortex is generally involved in processing complex events, such as action knowledge. Actions are represented in memory as sequences of goal-directed discrete events. Complex events can be divided into discrete smaller units and organised in a hierarchical manner. The aim of the present study was to investigate the ability of 9 patients with prefrontal lesion and 12 healthy controls to parse action sequences into meaningful events. Subjects were requested to detect the transitions between events under three different orientation instructions: (1) spontaneous; (2) small events; and (3) large event. Both normal subjects and patients identified significantly more events under the small-oriented condition. However, contrary to normal controls (NCs), patients with prefrontal damage showed considerable difficulties in detecting large event units than small event units. These results strongly suggest that prefrontal cortex is specifically involved in parsing and recognising clusters of action sequences. These findings also show that the way in which instructions orient subjects' attention, has an effect on how action information is encoded and represented in memory. PMID- 12887988 TI - The dependence of ipsilesional aiming deficits on task demands, lesioned hemisphere, and apraxia. AB - Neuroimaging studies as well as neurophysiological and lesion data indicate that the ipsilateral hemisphere plays a role in controlling the active limb. However, the nature and the conditions of this ipsilateral control are not well understood. We measured aiming movements with the ipsilesional limb toward targets with different characteristics which were made by patients with unilateral left brain damage (LBD) or right brain damage (RBD). The movement kinematics were analysed. Performance measures of the pointing movements were impaired in LBD patients, whereas RBD patients performed normally. LBD patients had obvious deficits during all tasks; however, they were exacerbated when high accuracy was required, and when an exocentric target had to be reached without visual feedback. Thus, the motor-dominant hemisphere plays a specific role in the programming and execution of ipsilateral aiming movements, and the importance of ipsilateral control increases with increasing task demands. To assess the relationship between pointing deficits and apraxia in LBD patients, the imitation of meaning gestures was tested. We replicated a recent study, showing that deviations of the final hand position from the demonstration were not correlated with abnormal kinematics of the corresponding arm movement when LBD patients performed this test. However, there were correlations between related kinematic measures during pointing and gesture imitation. These findings suggest a deficit of motor programming and execution after damage to the motor-dominant brain which is unrelated to the spatial errors characteristic of apraxia. This deficit affects different types of goal-directed aiming movements and its severity depends on task demands. PMID- 12887989 TI - Surface dyslexia in a Japanese patient with semantic dementia: evidence for similarity-based orthography-to-phonology translation. AB - We studied the reading performance of a Japanese-speaking patient, TI, with bilateral but asymmetrical (left more than right) temporal-lobe atrophy, severe anomia, and poor word comprehension. Most Japanese kanji characters correspond to several different legitimate pronunciations in different contexts, with varying degrees of correspondence consistency. TI made many errors in reading aloud words that violate statistically typical character-sound correspondences, especially for less common words, but had relatively preserved ability to read aloud strings in which the assignment of the typical pronunciation for each component character yields the correct pronunciation for the whole word. The degree of consistency of character-sound correspondences affected his performance on both words and nonwords in a graded manner. One interpretation is that TI's surface dyslexic reading reflects intact direct computation of phonology from orthography, but without the additional constraint from word meaning that is, in this framework, considered critical for correct pronunciation of lower-frequency words with atypical character-sound correspondences. Another interpretation is that TI's performance reflects partially damaged lexical knowledge of whole-word orthography and phonology, coupled with spared sublexcal knowledge of character sound correspondence rules. Whichever of these interpretations is preferred, this study offers the most detailed information yet available on the characteristics of surface dyslexia in Japanese. PMID- 12887990 TI - Selective disturbance of mental rotation by cortical stimulation. AB - In order to plan activity, people must imagine the spatial consequences of potential actions. Two classes of mental spatial transformation can be distinguished: Object-based spatial transformations are imagined movements of objects, such as mental rotation. Egocentric perspective transformations are imagined changes in one's viewpoint, such as imagining one's self in the position of another person. Here we report a case in which electrical stimulation of the right parietal cortex selectively interfered with performance of a mental rotation task. Interference was selective to this stimulation site, and was task specific. Performance of the perspective transformation task, and a control for visual encoding and responding, were unimpaired by stimulation. This marks the first instance of the use of direct cortical stimulation to investigate mental spatial transformations. PMID- 12887991 TI - Frontal and parietal participation in problem solving in the Tower of London: fMRI and computational modeling of planning and high-level perception. AB - This study triangulates executive planning and visuo-spatial reasoning in the context of the Tower of London (TOL) task by using a variety of methodological approaches. These approaches include functional magnetic resonance imaging (fMRI), functional connectivity analysis, individual difference analysis, and computational modeling. A graded fMRI paradigm compared the brain activation during the solution of problems with varying path lengths: easy (1 and 2 moves), moderate (3 and 4 moves) and difficult (5 and 6 moves). There were three central findings regarding the prefrontal cortex: (1) while both the left and right prefrontal cortices were equally involved during the solution of moderate and difficult problems, the activation on the right was differentially attenuated during the solution of the easy problems; (2) the activation observed in the right prefrontal cortex was highly correlated with individual differences in working memory (measured independently by the reading span task); and (3) different patterns of functional connectivity were observed in the left and right prefrontal cortices. Results obtained from the superior parietal region also revealed left/right differences; only the left superior parietal region revealed an effect of difficulty. These fMRI results converged upon two hypotheses: (1) the right prefrontal area may be more involved in the generation of a plan, whereas the left prefrontal area may be more involved in plan execution; and (2) the right superior parietal region is more involved in attention processes while the left homologue is more of a visuo-spatial workspace. A 4CAPS computational model of the cognitive processes and brain activation in the TOL task integrated these hypothesized mechanisms, and provided a reasonably good fit to the observed behavioral and brain activation data. The multiple research approaches presented here converge on a deepening understanding of the combination of perceptual and conceptual processes in this type of visual problem solving. PMID- 12887992 TI - Hypometric primary saccades and increased variability in visually-guided saccades in Huntington's disease. AB - Eye movement abnormalities can be distinctive and suggestive of a specific pathophysiology. To further investigate the deficits in the control of saccades in patients with Huntington's disease (HD), we investigated the ability of 11 HD patients and 11 matched controls to perform visually-guided saccades. We adopted reflexive saccade tasks involving predictable and unpredictable sequences, at different amplitudes of target step (10 degrees, 20 degrees, 30 degrees, 40 degrees ), as well as voluntary self-paced saccades. Prolongation of initiation was observed in the HD group as the target amplitude of predictable saccades increased. During the self-paced saccade task, the HD patients had increased intersaccadic intervals, performed fewer saccades in the allocated time and displayed an increased temporal variability in comparison to the controls. Furthermore, hypometric primary saccades, and an increased number of corrective saccades, were observed during both reflexive and voluntary saccades in the HD group. The delayed initiation of large saccades, deficits in voluntary, self paced saccades, impaired saccadic accuracy and increased corrective saccades in HD, were interpreted in light of other ocular motor and limb studies, and appear to be due to damage to the fronto-striatal loop, including the supplementary eye fields, as well as possible brainstem and cerebellar involvement. PMID- 12887993 TI - Assessing effects of stimulus orientation on perception of lateralized words and nonwords. AB - Numerous studies of the processes of visual word recognition in the left and right cerebral hemispheres have attempted to control for confounding differences in the retinal placement (and hence visual acuity) of the beginnings of words by re-orientating normally-horizontal words vertically. However, despite the popularity of this approach, little is known about the precise effects that vertically orientating normally-horizontal words exert on hemispheric processes of word recognition. In this study, we investigated perception of horizontal and vertical English words and nonwords in the left visual field (LVF) and right visual field (RVF). An eye-tracking device ensured central fixation and a 2AFC paradigm (Reicher-Wheeler task) suppressed influences of non-perceptual bias. Horizontal stimuli produced a strong right visual field advantage for words but not for nonwords, whereas, vertical stimuli produced no hemifield differences at all. Moreover, vertical stimuli produced an advantage for words over nonwords in both visual fields whereas horizontal stimuli produced this effect only in the right visual field. Implications of these findings for the sensitivity of processes of word perception to stimulus orientation in the two cerebral hemispheres are discussed. PMID- 12887994 TI - Perception of global facial geometry in the inversion effect and prosopagnosia. AB - We investigated how efficiently combinations of positional shifts in facial features were perceived and whether the effects of combinations on the overall geometry of the face were reflected in discriminative performance. We moved the eyes closer together or further apart, and moved the mouth up or down. Trials with combinations of changes to both the mouth and the eyes were contrasted with trials with single changes to either the mouth or the eyes. As a contrast, we also examined combinations of changes in eye colour (brightness) and the same spatial manipulations. In addition, we specifically contrasted spatial combinations that more severely distorted the original triangular relation of the mouth and eyes (e.g. eyes closer and mouth down) to those that better preserved the original aspect ratio (e.g. eyes farther and mouth down). This we termed the "geometric context effect". We found that combinations of two spatial changes were detected more quickly and accurately by normal subjects viewing upright faces but not when faces were inverted. In contrast, combinations of spatial shifts and eye colour changes showed no advantage over faces with only one type of change. Combinations of spatial changes that distorted overall facial geometry more were detected more efficiently than less distorting combinations, showing that the spatial shifts were perceived in the context of the global facial structure. Again, this was found for upright but not inverted faces. We also tested a prosopagnosic patient, who showed the advantage for two spatial changes over one but lacked this geometric context effect, implying that she did not integrate local spatial information into overall facial structure. PMID- 12887996 TI - Regional differences of type II collagen synthesis in the human temporomandibular joint disc: immunolocalization study of carboxy-terminal type II procollagen peptide (chondrocalcin). AB - The purpose of this study was to determine the regional differences of distribution of the carboxy-terminal type II procollagen peptide (pCOL-II-C; chondrocalcin) as markers of cartilaginous expression in the human temporomandibular joint (TMJ) disc. Twelve human TMJ discs without morphologic abnormalities were obtained from 12 fresh cadavers. All specimens were analysed for pCOL-II-C expression using polyclonal rabbit anti-human pCOL-II-C antibody in avidin-biotin-peroxidase complex staining. The results were demonstrated that the percentage of pCOL-II-C immunoreactive disc cells was significantly higher in the outer part (the articular surfaces) than in the inner part (the deep central areas) of the disc. These findings suggest that the tissue heterogeneity of cartilaginous expression reflects the functional demands of the remodelling process in the human TMJ disc. PMID- 12887997 TI - Antisense oligonucleotide against 47-kDa heat shock protein (Hsp47) inhibits wound-induced enhancement of collagen production. AB - It is well known that excessive collagen synthesis during the wound-healing process causes scar formation. Our recent in-vivo study indicates that antisense treatment against 47-kDa heat shock protein (Hsp47), a collagen-specific molecular chaperone, relieves scar formation following skin wounds in rats [Wang et al., Plast. Reconstr. Surg., in press]. In order to understand the mechanism of this phenomenon, we examined the effects of antisense treatment on the expression of mRNAs and proteins of Hsp47 and collagens in fibroblasts derived from wounded rat tongues. Hsp47 and procollagen alpha1(I) and alpha1(III) mRNAs were consistently increased after wounding and were maximal at day 5 post-injury. Treatment with antisense oligonucleotide against Hsp47 efficiently blocked the production of procollagen alpha2(I) and alpha1(III) proteins, but had little effect on their mRNA levels. Therefore, we conclude that antisense oligonucleotide against Hsp47 inhibits the production of procollagen type I and III proteins in fibroblasts derived from wounded tongues, overcoming the increase in their mRNAs. PMID- 12887998 TI - Propulsive appliance stimulates the synthesis of insulin-like growth factors I and II in the mandibular condylar cartilage of young rats. AB - Functional orthopedic appliances correct dental malocclusion partially by exerting indirect mechanical stimulus on the condylar cartilage, modulating growth and the adaptation of orofacial structures. However, the exact nature of the biological responses to this therapy is not well understood. Insulin-like growth factors I and II (IGF-I and IGF-II) are important local factors during growth and differentiation of several tissues, including cartilage. The aim of this study was to verify the mRNA and protein expression of IGF-I and IGF-II in the condylar cartilage of young male Wistar rats that used a mandibular propulsive appliance for 3, 5, 7, 9, 11, 13 or 15 days. For this purpose, sagittal sections of decalcified and paraffin-embedded condyles were submitted to immunohistochemistry and in situ hybridization. IGF-I and IGF-II expression increased with developmental age in the control and treated rats. After 9 days of treatment the positivity for both peptides in the animals that wore the propulsive appliance increased even more, expressively different from the age matched controls. The expression patterns of both IGFs were similar, although IGF I labelling was stronger. Furthermore, the enhanced expression of both peptides was in parallel with the proliferating cell nuclear antigen (PCNA) positivity, a proliferation cell marker. The modulation of IGF-I and IGF-II expression in the condylar cartilage in response to the propulsive appliance suggests that both peptides are involved in the mandibular adaptation during this therapy. PMID- 12887999 TI - Regulation of gene expression of tumour necrosis factor-alpha by protein kinase C in the rat dental follicle. AB - Tooth eruption requires alveolar bone resorption to form an eruption pathway. Recent studies suggest that tumour necrosis factor-alpha (TNF-alpha) may increase bone resorption by promoting the recruitment of mononuclear cells to the dental follicle to form osteoclasts. Although the major osteoclast burst is seen early postnatally in the rat (day 3), the second round of minor osteoclastogenesis is around postnatal day 10. We have previously reported that TNF-alpha is expressed in the dental follicle of newborn rats with maximum expression at day 9. Such expression is enhanced by IL-1alpha in cultured dental follicle cells. In this report, regulation of TNF-alpha expression by protein kinase C (PKC) was studied both in vitro and in vivo. Incubating dental follicle cells with phorbolmyristate acetate (PMA), a PKC activator, significantly up-regulated TNF-alpha gene expression in a dosage-dependent manner. A PKC specific inhibitor, Go 6983, abolished this PMA effect on up-regulation of TNF-alpha, but had no effect on IL 1alpha induced expression. TNF-alpha expression was significantly greater after treatment with a combination of PMA and IL-1alpha than in treatments with PMA or IL-1alpha alone, suggesting a synergistic effect on enhancing TNF-alpha expression. These gene expression results were confirmed at the protein level by immunostaining for TNF-alpha in the dental follicle cells. In vivo, injection of PMA into postnatal rats also increased TNF-alpha expression. Thus, PKC up regulates TNF-alpha expression in dental follicle cells, as does IL-1alpha. However, they appear to utilize different pathways to regulate TNF-alpha expression. PMID- 12888000 TI - Detection and characterization of a rat parotid gland protein kinase that catalyzes phosphorylation of matured destrin at Ser-2. AB - Destrin, an actin-binding protein, is partly phosphorylated at Ser-2 (numbering on the matured form) in the resting rat parotid gland, and beta-adrenergic or cholinergic stimulation of this gland induces its dephosphorylation. In this study, we searched for the protein kinase involved in phosphorylation of destrin. We developed an assay method for the kinase, using an antibody specific to destrin phosphorylated at Ser-2, and detected the kinase in the rat parotid homogenate. This enzyme was predominantly (93%) present in the soluble fraction, and the enzyme in this fraction was characterized. It had an optimum pH at 6.8 and required 3-5 mM Mg2+ for its maximum activity. Ca2+ (1 mM) had no effect whereas Mn2+ (5 mM) inhibited the enzyme activity by 75%. The apparent Km values for destrin and ATP were 92 microg/ml and 170 microM, respectively. GTP was an inefficient phosphate donor, and cAMP had no effect. Heat-denatured destrin was poorly phosphorylated. Two-dimensional PAGE analysis of destrin phosphorylated with the soluble fraction indicated that it was exclusively phosphorylated at Ser 2. None of the protein kinase inhibitors tested here was specific to this enzyme. At 1 mM, ML-7, Y-27632, KN-93, HA-1077, H-7, and H-8 inhibited the activity by 88, 61, 58, 49, 46, and 42%, respectively. Staurosporine (2 microM) and H-89 (50 microM) inhibited the activity by 48 and 33%, respectively. Heparin (30 microg/ml) had no effect. These results suggest that the rat parotid gland contains a novel, constitutively active, soluble protein kinase catalyzing specific phosphorylation of destrin at Ser-2. PMID- 12888001 TI - Genetic contributions to expression of the baboon cingular remnant. AB - Primitive mammalian molar morphology is characterised in part by a ridge of enamel that encircles the entire base of the molar crown, the cingulum. Many higher primates have reduced the cingulum, but often retain remnant features on the lingual surface of maxillary molars and the labial surface of mandibular molars. Two of these remnants in cercopithecoid primates, the interconulus and interconulid, are morphologically similar though the interconulus is found on maxillary molars and the interconulid is located on mandibular molars. Here we present results from a quantitative genetic analysis of expression of these two traits in a sample of 479 modern savannah baboons from the Southwest Foundation for Biomedical Research (SFBR). We found that both traits are significantly heritable with little variance attributable to other factors, such as sex, age, and molar crown size. Bivariate analyses yielded point estimates for genetic correlations between left and right side expression that are either equal to or not significantly different from 1.0; meaning that 100% of their additive genetic variance is due to the effects of the same gene or suite of genes. By contrast, our estimates of the genetic correlations between maxillary and mandibular expression of this trait range from 0.52 to 0.72, suggesting that 28-52% of the additive genetic variance in the interconulus and interconulid is due to the effects of shared genes. These results demonstrate that intra-arch expression is characterised by complete pleiotropy whereas inter-arch expression is caused by incomplete pleiotropy. These results are relevant to dental developmental studies as well as paleontological analyses of the evolution of the primate dentition. PMID- 12888002 TI - Tooth eruption and cementum formation in the Runx2/Cbfa1 heterozygous mouse. AB - Cleidocranial dysplasia (CCD) is an autosomal dominant human disorder that affects development of bones and teeth. The dental disorders in CCD patients include formation of supernumerary teeth, delayed tooth eruption, and lack of formation of cellular cementum in permanent teeth. This disorder involves a mutation in the osteoblast-specific transcription factor Runx2/Cbfa1, leading to haploinsufficiency of the Runx2/Cbfa1 protein. Here, we examined if Runx2/Cbfa1 heterozygous mice (with one functional allele for Runx2/Cbfa1) exhibit similar changes in tooth eruption, and dental cementum formation as in CCD patients. Heads of Runx2/Cbfa1 heterogeneous and wildtype mice aged days 16-35 postnatally were serially sectioned and stained with hematoxylin-eosin or for tartrate resistant acid phosphatase (TRAP) to identify osteoclasts. The results showed that the eruption pattern of the first and second molars in maxilla and mandible in Runx2/Cbfa1 +/- mice was the same as in wildtype animals. No clear difference in distribution or in the (estimated) number of osteoclasts was found. Cellular cement at the apical portions of the molar roots was present in both groups. The data suggests that in the mouse one allele for Runx2/Cbfa1 is sufficient for an undisturbed tooth eruption and an apparently normal formation of the periodontium. PMID- 12888004 TI - Dynamical modelling of pattern formation during embryonic development. AB - The combination of genetic and molecular biology techniques has uncovered the intricacies of several gene networks controlling developmental processes. In the face of such complex regulatory networks, developmental geneticists cannot rely on reasoning alone; a thorough understanding of the spatio-temporal properties of these networks clearly requires the use of proper computational tools and methods. PMID- 12888005 TI - Emerging systems: between vertebrates and arthropods, the Lophotrochozoa. AB - Novel molecular model organisms for the study of development and regeneration are emerging among the Lophotrochozoa, the third major branch of bilaterian animals. The polychaete Platynereis, the leech Helobdella, the snail Ilyanassa, and several planarians are efficiently accessed for molecular techniques including large-scale whole-mount in situ hybridization screening, RNA interference or morpholino knock-down. Joint efforts include the generation of genomic resources in the form of expressed sequence tag collections and bacterial artificial chromosome libraries. Current research focuses on early pattern formation during cleavage, the emergence and diversification of body segments, and the formation of photoreceptor cells and eyes. Several lophotrochozoan groups (in particular nereid polychaetes) exhibit modes of development, organ design, or body plans that are considered ancestral in many respects. This is also reflected in the level of genes, making these groups ideally suited for developmental comparative studies. PMID- 12888006 TI - Cell polarity: the ups and downs of the Par6/aPKC complex. AB - A signaling complex in which atypical protein kinase C associates with a regulatory protein, Par6, plays an essential role in establishing cell polarity. Recent studies in organisms ranging from worms to mammals have highlighted some of the conserved mechanisms by which the assembly, localization and activity of this complex are regulated. Recent work is also beginning to unravel how this complex acts in concert with additional molecular complexes to establish and maintain polarity. PMID- 12888007 TI - Regulation of membrane dynamics in developing epithelia. AB - A cellular understanding of developmental mechanisms embraces complex dynamic processes at the cell surface. Studies of trafficking pathways and of associated cytoskeletal elements in genetically tractable organisms allow a better analysis of the mechanisms underlying the control of cell polarization and tissue morphogenesis at different stages of animal development. It is now possible to identify signalling pathways that provide the temporal triggers and/or spatial cues to regulate core cellular effectors during migration, morphogenesis and the formation of epithelia. PMID- 12888008 TI - Chemotactic cell movement during development. AB - Chemotaxis is an important mechanism controlling cell migration over either short or long distances during different developmental processes. Small rapid diffusing chemo-attractants are detected through serpentine, G protein coupled receptors through graded activation of receptors along the length of the cell. Internal amplification results in polarisation of the actin and myosin cytoskeletal dynamics along the gradient and directed movement. The dynamics of these processes can now be studied in individual cells in developing organisms. Slow diffusing chemo-attractants such as growth factors, providing short-range guidance information, often signal through tyrosine kinase receptors. Detection of these signals may involve the active extension of very long cellular process up growth factor gradients, followed by translocation of the cell in the direction of the gradient. PMID- 12888009 TI - Cell adhesion in development: a complex signaling network. AB - Cell-adhesion molecules play a major role in morphogenesis and organogenesis. In vertebrates, a significant fraction of genes encode cell-adhesion molecules. Multiple signal-transduction pathways have been described that modulate the adhesion process. These pathways have been studied in great detail for cadherins and integrins - two major adhesion systems controlling cell-cell and cell substrate interactions. Recent findings confirm that a given cell-adhesion molecule can be implicated at different stages of development in processes as diverse as cell positioning, tissue patterning and compartmentalization, axon guidance and synaptogenesis. Clearly, a wide variety of new biophysical techniques and genomic approaches will permit analysis of the roles of adhesive interactions in development to be addressed with far greater precision. PMID- 12888010 TI - miSSING LINKS: miRNAs and plant development. AB - The discovery of hundreds of plant micro RNAs (miRNAs) has triggered much speculation about their potential roles in plant development. The search for plant genes involved in miRNA processing has revealed common factors such as DICER, and new molecules, including HEN1. Progress is also being made toward identifying miRNA target genes and understanding the mechanisms of miRNA-mediated gene regulation in plants. This work has lead to a reexamination of many previously characterized mutations that are now known to affect components or targets of miRNA-mediated pathways. PMID- 12888011 TI - Oscillations, clocks and segmentation. AB - Notch signalling molecules, such as the basic helix-loop-helix factors Hes1 and Hes7, periodically change their expression in the presomitic mesoderm, and each cycle of gene expression is associated with somite formation (every two hours in mouse). This cyclic expression is the manifestation of an intrinsic mechanism, called the segmentation clock, which is essential for coordinated somite segmentation. Interestingly, the oscillatory expression of Hes1 is observed in many cell types after serum stimulation, suggesting that this ultradian clock is not unique to presomitic mesoderm cells but widely distributed. This oscillation depends on the negative feedback loop, and once its promoter is constitutively activated, Hes1 seems to start oscillatory gene expression autonomously. Thus, Hes1 acts as a device that transduces a direct current of input into an alternating current, which ticks the hours in many biological systems. PMID- 12888012 TI - Cilia are at the heart of vertebrate left-right asymmetry. AB - Handed asymmetry of the shape and position of the internal organs is found in all vertebrates, and is essential for normal cardiac development. Recent genetic and embryological experiments in mouse embryos have demonstrated that left-right asymmetry is established by directional flow of extraembryonic fluid surrounding the node, which is driven by motile monocilia. PMID- 12888013 TI - Early endoderm development in vertebrates: lineage differentiation and morphogenetic function. AB - Gastrulation of the vertebrate embryo culminates in the formation of three primary germ layers: ectoderm, mesoderm and endoderm. The endoderm contributes to the lining of the gut and the associated organs. New components of the molecular pathway for endoderm specification have been identified in the zebrafish and Xenopus. In the mouse, the activity of orthologous factors is involved with the allocation and differentiation of the definitive endoderm. Morphogenetic interactions between the endoderm and the other germ layer derivatives are critical for the morphogenesis of head structures and organogenesis of gut derivatives. PMID- 12888014 TI - Pancreas specification: a budding question. AB - Much recent investigation has been carried out into the mechanisms by which the pancreas is specified from the early endoderm. Recent advances have highlighted important roles for retinoic acid and bone morphogenetic protein signalling in patterning the endoderm at late gastrulation. Subsequently, interactions with the endothelium of the aorta in the dorsal pancreas domain and lateral plate mesoderm in the ventral pancreas domain are the source of essential pancreas-inductive signals. Additionally, the transcription factor Ptf1a has been demonstrated to have a previously unappreciated role in distinguishing pancreas from surrounding duodenal fates. PMID- 12888015 TI - Vascular development and patterning: making the right choices. AB - The developing vascular system is regulated by a series of intrinsic and extrinsic signaling interactions that result in the formation and specialization of vessels that circulate blood and lymph around the body. Signaling molecules, such as vascular epithelial growth factors (VEGFs) and angiopoietins, that establish the cellular identity of the endothelial cells in the early embryo also play roles in establishing whether developing vessels will be arterial, venous or lymphatic. Genetic studies in zebrafish and mice have suggested a key role for Notch signaling, downstream of VEGF-A, in specifying arterial versus venous fate, and for VEGF-C and -D, along with the transcription factor, Prox1, in specifying lymphatic fate. PMID- 12888016 TI - Stem cells to tissue: molecular, cellular and anatomical heterogeneity in skeletal muscle. AB - Multiple waves of muscle precursors are released from skeletal muscle progenitor cells throughout developmental life, and this process is initiated in precise locations in the embryo. Skeletal muscle diversifies not only after the acquisition of muscle identity, but curiously heterogeneity is observed even in the stem cell population. Recent studies on cell lineage, cell fusion and the nature of post-natal satellite cells have expanded on our fundamental knowledge of the formation of this tissue, and how this tissue is replenished by resident and circulating regenerative stem cells during adult life. PMID- 12888017 TI - Mouse models of telencephalic development. AB - As the telencephalon first emerges from anterior neuroectoderm, signalling molecules and transcription factors combine to specify the identity and fate of cells in each of its regions. Studies of both naturally occurring and transgenic mutant mice have identified many genes that contribute to this process. The development of telencephalon and its regions is specified by signalling molecules produced at sites both surrounding and within the telencephalon. Different parts of the telencephalon express different combinations of transcription factors that control processes including proliferation, cell fate determination and migration in order to create the unique phenotype of each region. PMID- 12888018 TI - The freshwater planarian Schmidtea mediterranea: embryogenesis, stem cells and regeneration. AB - Planarians have been used as a model to study development and regeneration for more than 200 years. Research on these animals has traditionally focused on surgical and pharmacological manipulations. Recently, the dissection of planarians has become more molecular in nature. The isolation of thousands of expressed sequence tags and the introduction of in situ hybridizations, immunocytology, and RNA-mediated gene interference has opened the door to gene discovery and to the study of gene function in planarians during development and regeneration. These advances promise to shed mechanistic insight into basic biological attributes such as regeneration and stem-cell regulation. PMID- 12888019 TI - Spinal cord pattern generators for locomotion. AB - It is generally accepted that locomotion in mammals, including humans, is based on the activity of neuronal circuits within the spinal cord (the central pattern generator, CPG). Afferent information from the periphery (i.e. the limbs) influences the central pattern and, conversely, the CPG selects appropriate afferent information according to the external requirement. Both the CPG and the reflexes that mediate afferent input to the spinal cord are under the control of the brainstem. There is increasing evidence that in central motor diseases, a defective utilization of afferent input, in combination with secondary compensatory processes, is involved in typical movement disorders, such as spasticity and Parkinson's disease. Recent studies indicate a plastic behavior of the spinal neuronal circuits following a central motor lesion. This has implications for any rehabilitative therapy that should be directed to take advantage of the plasticity of the central nervous system. The significance of this research is in a better understanding of the pathophysiology underlying movement disorders and the consequences for an appropriate treatment. PMID- 12888020 TI - Thirty minutes mobile phone use has no short-term adverse effects on central auditory pathways. AB - OBJECTIVE: To investigate whether pulsed high-frequency electromagnetic field (pulsed EM field) emitted by a mobile phone for 30 min has short-term adverse effects on the human central auditory system. METHODS: We studied the auditory brainstem response (ABR), the ABR recovery function and middle latency response (MLR) before and after using a mobile phone for 30 min in 15 normal hearing volunteers. RESULTS: None of the 3 measures were affected by exposure to pulsed EM field emitted by a mobile phone for 30 min. CONCLUSIONS: Based on the ABR and MLR methods utilized in the study, we conclude that 30 min mobile phone use has no short-term adverse effects on the human auditory system. PMID- 12888021 TI - The long-term effects of antiepileptic drugs on the visual system in rats: electrophysiological and histopathological studies. AB - OBJECTIVE: We quantified the long-term effects of antiepileptic drugs (AEDs) on the visual system of rats using electroretinograms (ERGs) and visual evoked potentials (VEPs). METHODS: Twenty adult Sprague-Dawley rats were divided into 4 groups (n=5). Each animal was treated by monotherapy of phenytoin (PHT), valproic acid (VPA), zonisamide (ZNS) or physiological saline as control. The AEDs were injected intraperitoneally daily for 180 days. ERGs and VEPs were recorded before the medication and on Days 30 and 180. RESULTS: There were no significant changes in the 4 groups on Day 30. On Day 180, the amplitudes of a- and b-waves of dark adapted (DA) ERGs were reduced in the PHT group compared with those of the control group. In the VPA group, the amplitudes of the DA ERG a- and b-waves, light-adapted ERG b-wave and the DA VEP were reduced. No significant changes were observed in the ZNS group. There were no histopathological changes of the retina and visual cortex in all groups. CONCLUSIONS: Our results indicate that neurons along the visual pathways have different sensitivity to each AED. This may result from the differential pharmacological actions of each AED on visual neurons. SIGNIFICANCE: Our findings suggest that epileptic patients on long-term use of AEDs might have subclinical influences to the visual systems. PMID- 12888022 TI - Intracranial volume conduction of cortical spikes and sleep potentials recorded with deep brain stimulating electrodes. AB - OBJECTIVE: To examine interictal epileptiform and sleep potentials recorded intracranially from deep brain stimulation (DBS) electrodes in patients treated with DBS for epilepsy. Specifically, this study sought to determine whether the DBS-recorded potentials represent: (a) volume conduction from surface neocortical discharges or (b) transsynaptic propagation along cortical-subcortical pathways with local generation of the subcortical potentials near the DBS targets. METHODS: Six patients with intractable epilepsy treated with thalamic DBS of the central median nucleus (CM; one patient) or anterior thalamus (5 patients) who had focal interictal spikes were studied. Sleep potentials were also studied in a 7th patient with Parkinson disease treated with DBS of the subthalamic nucleus (STN). RESULTS: Focal interictal cortical spikes recorded by scalp electroencephalography (EEG) were recorded synchronously, but with opposite polarity, from the DBS electrodes in CM as well as the more superficial anterior thalamic contacts situated in the anterior nucleus (AN) and dorsal medial nucleus (DM). In referential montages, the subcortical potentials were of highest amplitude ipsilateral to the focal cortical spikes, with a small but reproducible amplitude decrement present at each electrode contact more distant from the cortical source, irrespective of the specific DBS target. Subcortical sleep potentials (K-complexes and sleep spindles) were also recorded synchronously and with inverse polarity compared to the corresponding scalp potentials, and appeared in a similar fashion at all subcortical sites sampled by the DBS electrodes. Amplitude attenuation in the thalamus of intracranial volume conducted potentials with increasing distance from their cortical spike sources was measured at approximately 5-10 microV/mm. DISCUSSION: Recent reports on scalp CM or scalp-STN EEG recordings in patients treated with DBS for epilepsy have interpreted the intracranial waveforms as evidence of transsynaptic cortical subcortical transmission across neuroanatomical pathways presumed to be involved in the generation of sleep potentials (Clin. Neurophysiol. 113 (2002) 25) and epileptiform activity (Clin. Neurophysiol. 113 (2002) 1391). However, our results show that the intracranial spikes recorded from DBS electrodes in various regions of the thalamus (CM, AN and DM) represent subcortical volume conduction of the synchronous cortical spikes recorded with scalp EEG. The same is true for the intracranial reflections of scalp EEG sleep potentials recorded from DBS electrodes in CM, AN, DM and STN. These interictal DBS waveforms thus cannot be used to support hypotheses of specific cortical-subcortical pathways of neural propagation or subcortical generation of the DBS-recorded potentials associated with scalp EEG interictal spikes and sleep potentials. SIGNIFICANCE: Detailed analysis of the intracranial potentials recorded from DBS electrodes in association with scalp EEG spikes and sleep discharges shows that the intracranial waveforms represent volume conduction from discharges generated in the neocortex and not, as has been suggested, locally generated activity resulting from cortical-subcortical neural propagation. PMID- 12888023 TI - Early diagnosis of diabetic neuropathy using double-shock stimulation of peripheral nerves. AB - OBJECTIVE: The purpose of this study was to determine the changes in the amplitudes of a sensory nerve action potential (NAP) to a conditioning stimulus given prior to a test stimulus at 2-8 ms intervals in healthy subjects and patients with diabetes mellitus with no clinical signs of neuropathy and normal nerve conduction velocities (NCVs), to be able to diagnose peripheral neuropathy at its very early stages. METHODS: NAPs in the superficial branch of the radial nerve were recorded in healthy subjects (28 women and 7 men) and type II diabetes patients without neuropathy (22 women and 12 men). Radial nerve was first stimulated with a single shock and then with double shocks at intervals of 2, 3, 4, 5, 6, 7, and 8 ms; NAP amplitudes and NAP1/NAP2 ratios were calculated in normals and diabetics. NCVs were within the normal ranges (>50 m/s) in all subjects. RESULTS: Of the independent variables--group (control, patient), sex (male, female), and hand (right, left)--only group significantly influenced NAP amplitude; mean NAP amplitude (single shock) was significantly lower in patients than controls. NAP1/NAP2 ratios were slightly below one (facilitation) in controls; it was above one at 1-8 ms stimulus intervals (inhibition) in diabetics, which was strongest at smallest intervals, gradually decreasing, and almost disappearing as the stimulus interval approached 8 ms. CONCLUSIONS: Using double-shock stimuli, an early diagnosis of peripheral neuropathy would be possible in diabetics without clinical signs of peripheral neuropathy and exhibiting no slowing in NCV. PMID- 12888024 TI - Influence of stimulus intensity on waveform of sympathetic skin response evoked by magnetic stimulation. AB - OBJECTIVE: The aim of this study was to investigate the influence of stimulus intensity on the waveform of sympathetic skin response (SSR). The origin of the SSR waveform was discussed. METHODS: A total of 12 SSRs from palm skin were analyzed in 40 normal subjects. SSR was evoked by magnetic stimulation to the neck at 3 different intensities (10, 35, and 70% of the maximum output). The 3 stimuli were repeatedly applied in 4 separate sets. Waveforms were classified as either the P type, in which the positive component was larger than the negative one, or the N type, in which the negative component was larger than the positive one. Amplitude values of peak-to-peak, the first negative (N1), and subsequent positive (P1) components were compared among the 12 responses. RESULTS: When the stimulation was increased, the SSR size became larger. The P type SSR was most frequently found in the maximum stimulation. Strong stimulation generally produced a large P1. Only the N type SSR exhibited a large N1 response to the maximum stimulation. There was a significant, negative correlation between N1 peak duration (difference between the SSR onset and N1 peak latencies) and P1 amplitude. CONCLUSIONS: These results suggested that strong responses probably produced a P type SSR with a large P1 component. In this condition, a rapid change of potential from negative to positive prevented any correlation between N1 amplitude and the magnitude of the response. In the N type SSR, however, a large N1 was associated with a strong response. These findings can be explained by a newly presented theory on the neurophysiological origin of the negative and positive components, based on a model of equivalent current dipole dependent on the Na+ concentration gradient. SIGNIFICANCE: The present study provided some suggestions on the neurophysiological mechanism of SSR waveform. PMID- 12888025 TI - Relationships between the changes in compound muscle action potentials and selective injuries to the spinal cord and spinal nerve roots. AB - OBJECTIVE: Compound muscle action potentials (CMAPs) evoked by transcranial electrical stimulation have been widely introduced to monitor motor function during spinal surgery. They may reflect segmental injuries as well as injuries to motor-related tracts in the spinal cord. However, we have experience with some patients who developed postoperative segmental motor weakness without any potential changes during surgery. To evaluate the efficacy of this method, we used a cat model to observe the relationships between potential changes and selective injuries to the white and gray matters of the spinal cord and spinal nerve roots. METHODS: Ten CMAPs were obtained before and after injury to the spinal cord and spinal nerve roots in 20 cats. Changes in the amplitude, latency, and duration of CMAPs were analyzed. RESULTS: CMAPs decreased in amplitude significantly after the insult to the motor-related tracts in the spinal cord in all cats, while the potentials did not always change when the insult was restricted to a limited area in the anterior horn of the spinal cord or to the single spinal nerve root. CONCLUSIONS: CMAPs may not exactly reflect segmental injury, and careful attention should be paid to the interpretation of CMAPs. PMID- 12888026 TI - Changes in motor cortical excitability induced by paired associative stimulation. AB - OBJECTIVE: Changes in afferent input have been shown to be capable of inducing reorganisations of motor cortex in humans. Using TCMS we examined the efficacy of a new associative afferent stimulation paradigm in inducing motor cortical reorganisation in humans. METHODS: Using TCMS, stimulus response curves were constructed before and following a 1 h period of associative stimulation of two muscles motor points. The effect of an asynchronous peripheral stimulation paradigm was investigated in a separate series of control experiments. RESULTS: One hour of associative stimulation of two muscles motor points resulted in a significant increase in the excitability of the corticospinal projection to those stimulated muscles. The increase in excitability peaked 1 h following the stimulation period. This increase in excitability did not generalise to either adjacent or more remote muscles. The control stimulation paradigm produced no significant change in corticospinal excitability. CONCLUSIONS: These results confirm the importance of associative input for the induction of plasticity in the human motor cortex. SIGNIFICANCE: The findings reported here further elucidate the role of afferent input in motor cortical reorganisation. These findings have implications for our understanding of the mechanisms of motor learning and may also be relevant to the design of new afferent stimulation therapies. PMID- 12888027 TI - Finger interactions studied with transcranial magnetic stimulation during multi finger force production tasks. AB - OBJECTIVE: We used transcranial magnetic stimulation (TMS) to investigate interactions among neuromuscular complexes involved in force production by individual fingers of a hand during single- and multi-finger tasks. METHODS: Subjects were asked to press with the fingertips at various levels of force using different finger combinations. TMS was applied over the M1 cortical hand area during constant force production. TMS-induced increments in fingertip forces were analyzed, as well as motor evoked potentials (MEPs) in flexor digitorum superficialis. RESULTS: Both MEP size and individual force increments had inverted U-shaped dependences on the background force (peak responses were seen at about 50% of the maximal force). Similar relationships were obtained when subjects were asked to produce different forces with the same finger combination or the same total force with different finger combinations. The relationships were similar when the force was produced by explicitly instructed fingers or by other fingers of the hand. Effects of TMS on the force of a finger showed a strong dependence on the background force produced by this finger and minimal or no dependence on forces produced by other fingers of the hand. CONCLUSIONS: Overall, TMS applied over M1 revealed little interaction among fingers. This supports the notion of digit-specific compartments in multi-digit extrinsic muscles of the hand and suggests that these compartments possess a high degree of physiological independence. PMID- 12888028 TI - EMG responses evoked by the termination of galvanic (DC) vestibular stimulation: 'off-responses'. AB - OBJECTIVE: Vestibular responses in soleus electromyography (EMG) evoked by the sudden onset of galvanic (DC) stimulation ('on-responses') have been described in detail previously. The aim of the present study was to describe responses in soleus triggered by the termination of galvanic stimulation ('off-responses'). METHODS: In 10 healthy human subjects, we studied responses to transmastoid (bilateral) stimuli of 200 ms and 2 s average duration and 3 or 4 mA intensity. We obtained both on- and off-responses using the same raw data. EMG activity was recorded onto tape while current pulses of systematically varying duration were delivered. Averaged on-responses were obtained by triggering from the beginning of the current pulses. Averaged off-responses were obtained by triggering from the termination of the current pulses. RESULTS: Short-latency (SL) and medium latency (ML) off-responses were both obtained in all but one study. The SL and the ML components of the off-responses were present and had similar latencies and amplitudes, but opposite excitability, to the on-responses obtained with the same stimuli. CONCLUSIONS: Off-responses to galvanic vestibular stimulation can be recorded from soleus EMG. Our findings imply that vestibular SL and ML reflex responses in the legs are dependent on the change in the rate of vestibular nerve discharge, not its absolute level. Both on- and off-responses have properties appropriate to a role in maintaining body stability. PMID- 12888029 TI - Dynamic synchronisation of central oscillators in essential tremor. AB - OBJECTIVE: Coherence analysis of electromyography (EMG) signals in essential tremor (ET) suggests that tremor in the right and left arm is induced by independent central oscillators. The sensorimotor cortex seems to be part of the tremor-generating neuronal network in ET. Here, we investigated using electroencephalography (EEG) whether the independence of central oscillators in ET is supported by the analysis of cortical activity. METHODS: In 8 patients with ET, bilateral hand tremor was activated by wrist extension. EMGs from the wrist flexors and extensors were recorded simultaneously with an EEG. EEG-EMG coherence was estimated for 74 epochs of 60 s duration. RESULTS: In 42.6% of the cases, EEG EMG coherence at the tremor frequency existed only with the contralateral sensorimotor cortex. However, 21.6% of the tremor-EMGs were coherent with EEG activity over both the contralateral and ipsilateral sensorimotor cortex. Bilateral and exclusively contralateral EEG-EMG coherence could alternate within the same recording. Bilateral EEG-EMG coherence was associated with increased right-left EEG-EEG coherence, increased right-left EMG-EMG coherence as well as with increased tremor strength. CONCLUSIONS: In ET, central oscillators in the right and left brain are not entirely independent of each other. They may dynamically synchronise, presumably by interhemispheric coupling via the corpus callosum. PMID- 12888030 TI - Control of hand movements after striatocapsular stroke: high-resolution temporal analysis of the function of ipsilateral activation. AB - OBJECTIVE: Hemiparesis due to infarction of the middle cerebral artery has become an increasingly important focus of research on cortical plasticity. Positron emission tomography and functional magnetic resonance imaging studies in such patients found involvement of the hemisphere ipsilateral to the affected hand related to movements of this hand. To understand the function of this ipsilateral activation, the present study investigated movement-related electroencephalogram (EEG) potentials in patients and healthy control subjects to measure timing of ipsi- and contralateral activation relative to movement onset. METHODS: Thirteen patients were investigated in their chronic stage. Their pyramidal tracts were affected by infarctions of the middle cerebral artery at striatocapsular level. EEG potentials were recorded from 26 scalp electrodes while patients were pressing a key with their right or left index finger within a warned choice response task. RESULTS: Beginning 200 ms before responses of the affected hand, there was normal contralateral preponderance of EEG negativity. Briefly after response onset, however, the other unaffected hemisphere, ipsilateral to the responding hand, became additionally active. This pattern did not occur with responses made by the unaffected hand nor in healthy participants. CONCLUSIONS: The timing of the onset of ipsilateral activity precludes its role in response initiation. Rather, this activity may indicate reflex-like activation of the unaffected motor system to compensate for possible failure of the affected hand. PMID- 12888031 TI - Changes in median nerve somatosensory transmission and motor output following transient deafferentation of the radial nerve in humans. AB - OBJECTIVE: To determine if transient anaesthetic deafferentation of the radial nerve would lead to alterations in processing of early somatosensory evoked potentials (SEPs) from the median nerve or alter cortico-motor output to the median nerve innervated abductor pollicis brevis (APB) muscle. METHODS: Spinal, brainstem, and cortical SEPs to median nerve stimulation were recorded before, during and after ipsilateral radial nerve block with local anaesthesia. Motor evoked potentials (MEPs) and motor cortex output maps were recorded from the APB muscle. RESULTS: There were no significant changes to most early SEP peaks. The N30 peak, however, showed a significant increase in amplitude, which remained elevated throughout the anaesthetic period, returning to baseline once the anaesthetic had completely worn off. MEP amplitude of the median nerve innervated APB muscle was significantly decreased during the radial nerve blockade. There was also a significant alteration in the APB optimal site location, and a small but significant decrease in the silent period during the radial nerve blockade. CONCLUSIONS: Transient anaesthetic deafferentation of the radial nerve at the elbow leads to a rapid modulation of cortical processing of median nerve input and output. These changes suggest an overall decrease in motor cortex output to a median nerve innervated muscle not affected by the radial nerve block, occurring concomitantly with an increased amplitude of the median nerve generated N30 SEP peak, thought to represent processing in the supplementary motor area (SMA). Independent subcortical connections to the SMA are thought to contribute to the N30 response observed in this study. Unmasking of pre-existing but latent cortico cortical and/or thalamo-cortical connections may be the mechanism underlying the cortical SEP increases observed following radial nerve deafferentation. SIGNIFICANCE: Transient deafferentation of the radial nerve, which supplies wrist and hand extensor muscles, has been shown to alter sensory processing from and motor output to the median nerve innervated thenar muscles. PMID- 12888032 TI - Intracerebral somatosensory event-related potentials: effect of response type (button pressing versus mental counting) on P3-like potentials within the human brain. AB - OBJECTIVE: To assess the contribution of different anatomical brain sites to the genesis of P3 phenomena with respect to button pressing versus mental counting tasks. METHODS: Eight intractable epileptic patients undergoing depth electrode recordings prior to their surgery participated in the study. A total of 546 cerebral sites were recorded. A standard somatosensory oddball paradigm was used. The experiment was carried out in two sessions, differing in the requested responses to targets. The averaged responses in both tasks were compared. RESULTS: After targets, two kinds of P3-like potentials were observed within different cortical sites. Task-non-specific P3 potentials were seen for both types of responses to targets. The mean amplitude of these task-non-specific P3 potentials was significantly higher in the button pressing task. The intracerebral generators of this somatosensory P3 did not differ from the known generators of auditory and visual P3s. Task-specific P3-like potentials were found much less frequently. Button pressing unequivocally generated additional P3 like potentials in the premotor cortical sites. Mental counting repeatedly evoked additional P3-like waveforms in the left-side middle and inferior temporal gyri. CONCLUSIONS: In addition to multiple intracerebral P3 generators that reflect target detection processing, other task-specific P3-like potential generators can be found in the human brain. Their activity may affect the topography and precise parameters of scalp P3 potential. PMID- 12888033 TI - The disruptive effect of chronic pain on mismatch negativity. AB - OBJECTIVE: To investigate the effect of chronic pain on processes that generate the mismatch negativity (MMN). METHODS: Twelve participants with a diagnosis of chronic intractable pain were tested before and after pain treatment. During testing, event-related potentials were recorded while participants performed tasks of varying difficulty. RESULTS: The amplitude of the MMN was found to be greater following a nerve block procedure compared to MMN amplitude when participants were experiencing chronic pain. This effect was found to occur in the MMN for difficult-to-detect tones elicited while participants were performing a simultaneous cognitively demanding visual task. MMN amplitude was found to be greater with attention to difficult-to-detect deviants during pain but not in no pain conditions. CONCLUSIONS: These results provide an electrophysiological correlate of previous findings that high levels of pain disrupt cognition during the performance of demanding tasks. PMID- 12888034 TI - Event-related potentials associated with second language learning in children. AB - OBJECTIVE: In this study, we investigated learning-related changes in auditory event-related potentials (ERPs) of Finnish-speaking 3-6-year-old children caused by learning French language. METHODS: Using an oddball paradigm, ERPs to sounds of French language were recorded in the two groups of healthy children: those who were learning French (experimental group) and those who were not learning any foreign language (control peers). RESULTS: When the children from the experimental group were exposed to the foreign language, they automatically developed French-specific memory traces that helped them to discriminate, categorize, and pronounce utterances of the new language as indicated by the mismatch negativity (MMN) component of the ERPs in a previous study. We found that the learning process was also reflected by changes in P3a and late difference negativity (LDN) responses. Unlike MMN and P3a, the LDN has been discovered relatively recently and its functional role remains unclear. Similarly, as the MMN magnitude increased during the learning process, an increase of the P3a (known to reflect the involuntary attention switching toward deviant stimuli) and LDN amplitudes was observed. The ERPs of the control peers did not change significantly over the test period. CONCLUSIONS: When phonemes of a foreign language are learned, this process is accompanied with the increase in the MMN, P3a, and LDN amplitudes in children. Though the functional significance of LDN remains to be further investigated, our results support its possible link to reorienting processes following distraction. PMID- 12888035 TI - Differential characteristics of the middle latency auditory evoked magnetic responses to interstimulus intervals. AB - OBJECTIVE: The human middle latency auditory evoked magnetic fields were recorded with different interstimulus intervals (ISI) to investigate the differential natures of P30m and the P50m, including whether the P50m source was spatially different or not from the P30m source. METHODS: Twenty right-handed healthy subjects participated in the experiment. Auditory magnetic responses were recorded in the 0.5 s ISI (ISI were between 0.4 and 0.6 s) and the 1.5 s ISI conditions (ISI were between 1 and 2 s). Tone bursts were presented to the right ears 880 times consecutively for each condition. The P30m and the P50m responses were investigated, and the dipole source localization was performed. RESULTS: The P50m latency was significantly prolonged, while the P30m latency did not vary in the shorter ISI. Both P50m and P30m amplitudes were significantly reduced in the shorter ISI. The P50m was located significantly more anteriorly than P30m. CONCLUSIONS: These results suggest the existence of differential characteristic and spatially different magnetic responses in the middle latency range. SIGNIFICANCE: This study has revealed one aspect of the different natures between P30m and P50m, and may provide a key for auditory perceptional processes in humans. PMID- 12888036 TI - Repetitive transcranial magnetic stimulation of the parietal cortex transiently ameliorates phantom limb pain-like syndrome. AB - OBJECTIVE: Phantom pain is linked to a reorganization of the partially deafferented sensory cortex. In this study we have investigated whether the pain syndrome can be influenced by repetitive transcranial magnetic stimulation (rTMS). METHODS: Two patients with a longstanding unilateral avulsion of the lower cervical roots and chronic pain in the arm were studied. As a control the acute effects of rTMS (15 Hz, 2 s duration) on pain were studied in 4 healthy subjects. Pain intensity was assessed with the Visual Analogue Scale. RESULTS: Stimulation of the contralateral parietal cortex led to a reproducible reduction in pain intensity lasting up to 10 min. Stimulation of other cortical areas produced only minor alterations in the severity of the pain. Both 1 and 10 Hz rTMS trains applied to the contralateral parietal cortex on weekdays for 3 consecutive weeks did, however, not lead to permanent changes in the pain intensity. Experimentally induced pain (cold water immersion of the right hand) in normal subjects was not influenced by rTMS. CONCLUSIONS: These results do not favor the use of rTMS in the treatment of phantom limb pain. The results, however, support the concept that phantom pain is due to a dysfunctional activity in the parietal cortex. The transient rTMS-induced analgesic effect may be due to a temporary interference with the cerebral representation of the deafferented limb. PMID- 12888037 TI - Rapid reversible changes to multiple levels of the human somatosensory system following the cessation of repetitive contractions: a somatosensory evoked potential study. AB - OBJECTIVE: Numerous somatosensory evoked potential (SEP) studies have provided clear evidence that during repetitive voluntary movement, the transmission of somatosensory afferent information is attenuated. The objective of this work was to determine if this gating phenomenon could persist beyond the period of repetitive movement. METHODS: We recorded spinal, brainstem, and cortical SEPs to median nerve stimulation before and immediately after a modified 20 min repetitive typing task that did not involve the thenar muscles. RESULTS: There were significant decreases in pre-central cortical and subcortical SEP amplitudes for several minutes following task cessation. CONCLUSIONS: These results demonstrate the persistence of the gating phenomenon beyond the cessation of the actual repetitive movement. They also indicate that plastic changes do occur in cortical and subcortical components of the somatosensory system, following voluntary repetitive contractions. SIGNIFICANCE: The persistence of changes in somatosensory processing beyond the period of repetitive activity may be relevant to the initiation of overuse injuries. PMID- 12888038 TI - Influence of cholinergic circuitries in generation of high-frequency somatosensory evoked potentials. AB - OBJECTIVE: High-frequency oscillations (HFOs) evoked by upper limb stimulation reflect highly synchronised spikes generated in the somatosensory human system. Since acetylcholine produces differential modulation in subgroups of neurons, we would determine whether cholinergic drive influences HFOs. METHODS: We recorded somatosensory evoked potentials (SEPs) from 31 scalp electrodes in 7 healthy volunteers, before and after single administration of rivastigmine, an inhibitor of central acetylcholinesterase. Right median nerve SEPs have been analysed after digital narrow bandpass filtering (500-700 Hz). Raw data were further submitted to Brain Electrical Source analysis (BESA) to evaluate the respective contribution of lemniscal, thalamic and cortical sources. Lastly, we analysed by Fast Fourier transform spectral changes after drug administration in the 10-30 ms latency range. RESULTS: Rivastigmine administration caused a significant increase of HFOs in the 18-28 ms latency range. Wavelets occurring before the onset latency of the conventional N20 SEP did not show any significant change. A similar increase concerned the strength of cortical dipolar sources in our BESA model. Lastly, we found a significant power increase of the frequency peak at about 600 Hz in P3-F3 traces after drug intake. CONCLUSIONS: Our findings demonstrate that the cortical component of HFOs is significantly enhanced by cholinergic activation. Pyramidal chattering cells, which are capable to discharge high-frequency bursts, are mainly modulated by cholinergic inputs; by contrast, acetylcholine does not modify the firing rate of fast-spiking GABAergic interneurons. We thus discuss the hypothesis that cortical HFOs are mainly generated by specialised pyramidal cells. PMID- 12888039 TI - Spectral composition of NREM sleep in healthy subjects with moderately increased daytime sleepiness. AB - OBJECTIVE: This study addressed the relationship between daytime sleepiness and spectral composition of the preceding NREM sleep. METHODS: Nineteen healthy volunteers (mean age: 36.5 years; SD: 10.1) underwent polysomnography during two consecutive nights and the multiple sleep latency test (MSLT) on the following day. Daytime sleepiness was also assessed by the Epworth sleepiness scale (ESS). The sleep recordings were visually scored according to standard criteria. The quantitative sleep EEG analysis was performed using a fast Fourier transform routine. The sleep parameters were compared between subjects with short and long MSLT sleep latencies (cut-off=10 min) and between subjects with low and high ESS scores (cut-off=6 points). RESULTS: Subjects with short MSLT sleep latencies showed a reduced theta EEG activity. There was no evidence of reduced synchronization of sleep EEG in subjects with high ESS scores. CONCLUSIONS: Moderately increased daytime sleepiness as indicated by MSLT sleep latency less than 10 min is accompanied by decreased power of theta activity during NREM sleep indicating a deficit of sleep EEG synchronization. PMID- 12888042 TI - Internal limiting membrane staining with various concentrations of indocyanine green dye under air in macular surgeries. AB - PURPOSE: To determine the effective minimal concentration of indocyanine green (ICG) for staining the internal limiting membrane (ILM) under air in macular surgeries and to report the clinical outcome of these patients. DESIGN: Prospective, randomized clinical trial. METHODS: Consecutive cases of macular hole (17 cases) and epiretinal membrane (ERM) (11 cases) undergoing primary surgery with ICG-stained ILM peeling were randomized to receive one of the three concentrations (mg/ml) of ICG (1): 0.25, (2) 0.5, and (3) 1.25. The number of ICG injections, visual quality of the stained ILM, and time used for ILM peeling were recorded. Internal limiting membrane specimens were subsequently examined under electron microscopy. Preoperative and postoperative clinical data with fluorescein angiography were recorded. RESULTS: There was a significantly smaller number of eyes with poor ILM staining in the 1.25-mg/ml group compared with the 0.25- to 0.5-mg/ml group (Fisher exact test, P =.04). The mean time required for ILM peeling was 4.2 minutes (range, 2.0-8.1 minutes). There was no significant difference in the time required for ILM peeling among the three concentration groups (one-way analysis of variance, P =.18) or between the macular hole and ERM group (two-tailed t test, P =.34). No ICG toxicity was found clinically or angiographically, except in one suspected case with ERM formation at the edge of ILM peeling. Electron microscopy of ILM specimens did not reveal any retinal elements. CONCLUSIONS: 1.25-mg/ml ICG under air stains the macular ILM consistently well for its removal in macular surgeries. The safety of ICG-stained ILM peeling needs further evaluation. PMID- 12888043 TI - Extensive investigation of a large Brazilian pedigree of 11778/haplogroup J Leber hereditary optic neuropathy. AB - PURPOSE: To conduct systematic epidemiologic, neuro-ophthalmologic, psychophysical, and mitochondrial DNA (mtDNA) genetic examinations on a newly identified pedigree with Leber hereditary optic neuropathy (LHON). DESIGN: Observational population cohort study. METHODS: A prospective investigation of an entire Brazilian LHON family. SETTING: A field investigation by an international team conducted in a remote part of Brazil. STUDY POPULATION: We evaluated 265 (both eyes) of the 328 living family members of this LHON pedigree. Only members of this pedigree were studied. Those entering the pedigree as spouses were used as controls. OBSERVATION PROCEDURES: We conducted epidemiologic interviews emphasizing possible environmental risk factors, comprehensive neuro ophthalmologic examinations, psychophysical tests, Humphrey visual field studies, fundus photography, and blood testing for mitochondrial genetic analysis. RESULTS: We reconstructed a seven-generation maternal lineage descended from a common ancestor dating to the 1870s. All maternally related family members were invariably homoplasmic 11778 with a haplogroup J mtDNA, 33 being affected, of which 22 are still living. With each subsequent generation, there was a progressive decrease of penetrance, and only males were affected in the last two generations. A significant exposure (greater than 95% confidence intervals) to a variety of environmental risk factors characterized the affected individuals, with smoking as the most common (P <.01). Both affected and carriers (95% confidence intervals) presented with a significantly lower incidence of hypertension and high cholesterol compared with the control group (P <.05). CONCLUSIONS: Almost 95% of a 328-living-member pedigree with LHON 11778/J haplogroup was comprehensively studied. Our initial results indicate the strong influence of environmental risk factors. The remarkably reduced incidence of cardiovascular risk in the maternal lineage is discussed. Further genetic analysis may reveal a role for the nuclear genome. PMID- 12888044 TI - Lichen planus and cicatrizing conjunctivitis: characterization of five cases. AB - PURPOSE: To report the clinical and immunopathologic features and the response to therapy in a series of six patients with cicatrizing conjunctivitis due to lichen planus. DESIGN: Retrospective case series. METHODS: All six patients were seen in an ocular pemphigoid clinic. Clinical, immunopathologic, and serologic features were evaluated and therapeutic response in each patient was monitored. RESULTS: All six patients had evidence of conjunctival scarring. Five patients had lichen planus of the oral mucosa and gingiva; one patient had involvement of the skin. Histologic findings consisted of thickened epithelium and an interface lymphocytic infiltrate along the lamina propria. In three patients, electron microscopy of the conjunctiva revealed thickening, fragmentation, and duplication of the basement membrane zone. Direct immunofluorescence examination of the conjunctiva and oral mucosa demonstrated linear and shaggy fibrinogen deposition along the basement membrane zone, confirming the diagnosis of lichen planus. All six patients were placed on immunosuppressive therapy with control of the disease. However, only one patient was able to discontinue the anti-inflammatory medication and have the lichen planus remain in remission. CONCLUSIONS: Lichen planus should be included in the differential diagnosis of cicatrizing conjunctivitis. Performing appropriate investigations to distinguish conjunctival lichen planus from other autoimmune diseases such as mucous membrane pemphigoid is critical to managing the patient with cicatrizing conjunctivitis appropriately. Oral cyclosporine effectively controlled the conjunctival lichen planus in four of the six cases. PMID- 12888045 TI - Randomized clinical trial of topical betaxolol for persistent macular edema after vitrectomy and epiretinal membrane removal. AB - PURPOSE: To report the efficacy and safety of topical betaxolol for treatment of persistent macular edema. DESIGN: Randomized clinical trial. METHODS: Thirty seven eyes (37 patients) with best-corrected visual acuity between 20/200 and 20/50 and macular edema that remained for 3 months after vitrectomy and removal of epiretinal membrane were prospectively, randomly assigned to receive betaxolol or placebo. Nineteen eyes of 19 patients received betaxolol twice daily, and 18 eyes of 18 patients received placebo as a randomized comparison group. The patients were followed up for 6 months. This study evaluated the effect of betaxolol on best-corrected visual acuity and area of macular edema, which was digitally measured on serial fluorescein angiogram. Calculations of mean best corrected visual acuity were based on logarithm of the minimal angle of resolution (logMAR). To assess changes in area of edema, the initial (pretreatment) size of the edema was set to 100%, and all posttreatment measurements were normalized relative to the initial size. RESULTS: Mean best corrected visual acuity at baseline was 0.216 (20 of 92.6) and 0.244 (20 of 82.0) in the treatment and control group, respectively. Mean area of macular edema was 2.271 +/- 1.629 mm(2) and 2.273 +/- 1.209 mm(2) in the treatment and control group; there was no significant difference. The visual acuity at 6 months after the start of the follow-up was 0.471 (20 of 42.5) in the treatment group and 0.236 (20 of 84.7) in the control group. Mean changes in logMAR of visual acuity for 3- and 6-month follow-up were -0.282 +/- 0.191 and -0.337 +/- 0.197 in the treatment group, and -0.016 +/- 0.186 and +0.015 +/- 0.267 in the control group; a significant difference was found (P <.0001; P <.0001). Areas of macular edema at 6 months after the start of the follow-up were 1.492 +/- 1.357 mm(2) in the treatment group and 2.125 +/- 1.434 mm(2)in the control group. Mean change in area of the edema for 6 months were 76.5% +/- 24.1% and 63.4% +/- 28.3% in the treatment group and 92.9% +/- 15.4% and 87.4% +/- 25.6% in the control group; treated patients showed a significantly larger reduction than untreated patients at each examination (P =.0193; P =.0102). No complication associated with treatment or placebo was found. CONCLUSIONS: Topical betaxolol appeared to have a favorable treatment effect in eyes with macular edema that remained after vitrectomy and removal of epiretinal membrane. Further investigation of more cases and longer follow-up are needed. PMID- 12888046 TI - Visual field defects after uneventful vitrectomy for epiretinal membrane with indocyanine green-assisted internal limiting membrane peeling. AB - PURPOSE: To report visual field defects after vitrectomy for epiretinal membrane with indocyanine green (ICG)-assisted internal limiting membrane (ILM) peeling. DESIGN: Interventional consecutive case series. METHODS: A retrospective review of 16 eyes of 16 patients who underwent vitrectomy for idiopathic epiretinal membrane. Indocyanine green-assisted ILM peeling was performed in 7 of 16 eyes. The main outcome measure was postoperative visual field. RESULTS: Four of seven eyes (57%) with ICG-assisted ILM peeling had visual field defects postoperatively. The field defects identified were nasal in three eyes; in the fourth eye, the visual field was constricted to approximately 30 degrees. None of the nine eyes without ICG-assisted ILM peeling had a visual field defect. CONCLUSIONS: Peripheral visual field defects may occur after vitrectomy with ICG assisted ILM peeling. Although the cause of the defects is unclear, the potential role of ICG toxicity requires further investigation. PMID- 12888047 TI - Vitreous hemorrhage in patients with high-risk retinopathy of prematurity. AB - PURPOSE: To investigate outcomes in premature infants with high-risk retinopathy of prematurity and secondary vitreous hemorrhage. DESIGN: Retrospective chart review. METHODS: Patients were selected from a database of infants undergoing retinopathy of prematurity screening from September 1997 to November 1999. Infants with high-risk retinopathy of prematurity (zone I or posterior zone II threshold disease) with and without vitreous hemorrhage were compared. MAIN OUTCOME MEASURES: Final stage of retinopathy of prematurity and short-term structural outcome were assessed. Visual acuity and refraction were measured when possible. RESULTS: Twenty-two eyes of 11 patients (group 1) had high-risk (posterior zone II or zone I threshold) retinopathy of prematurity without vitreous hemorrhage. Group 1 patients had a 91% favorable short-term structural outcome. Eight eyes of five infants developed vitreous hemorrhage with high-risk retinopathy of prematurity (group 2). Group 2 patients had only a 12.5% favorable short-term structural outcome. Seven of eight (87.5%) progressed to stage IVa or IVb retinopathy of prematurity. Six eyes underwent vitreoretinal surgery after a median duration of hemorrhage of 36 +/- 29 days (4-70 days). Three eyes developed stage V detachments and three progressed to phthisical degeneration. Final visual acuity was no light perception in three eyes. CONCLUSION: Vitreous hemorrhage, in association with advanced retinopathy of prematurity, is a poor prognostic sign. PMID- 12888048 TI - Uveal metastasis from breast cancer in 264 patients. AB - PURPOSE: Breast cancer is an increasingly important health problem in women and is the most common tumor to metastasize to the uvea. This study was designed to evaluate the clinical features, management, and prognosis of patients with uveal metastasis from breast cancer. DESIGN: Retrospective interventional case series. METHODS: We retrospectively reviewed 264 consecutive patients with uveal metastasis from breast cancer. We assessed the clinical features of the patient and tumor at the time of presentation, management, and prognosis. Kaplan-Meier survival estimates were used to analyze the probability of death as a function of time. RESULTS: Uveal metastasis was the initial manifestation of breast cancer in seven patients (3%) and the first systemic metastatic site of previously diagnosed breast cancer in 43 (16%). Associated with uveal metastasis, optic disk metastasis was found in 13 patients (5%), eyelid metastasis in one patient (1%), and conjunctival and orbital in one patient (<1%). Of 264 patients with uveal metastasis, 225 (85%) had choroidal metastasis, eight (3%) iris metastasis, two (<1%) ciliary body metastasis, and 29 (11%) had metastasis in multiple uveal sites. In the 264 patients with uveal metastasis, the most common symptom was blurred vision in 197 patients (88%), floaters in 15 (5%), photopsia in 12 (5%), and 19 (7%) were asymptomatic. The uveal metastases were bilateral in 99 patients (38%) and unilateral in 165 (62%). In 55 (56%) of the 99 bilateral cases, a uveal metastasis was found in the asymptomatic fellow eye during follow-up examination. External beam radiotherapy was used in 137 patients with uveal metastasis (52%), providing tumor control in 116 patients (85%) at a mean follow-up of 21 months. Using Kaplan-Meier estimates, survival rates of all patients with uveal metastasis from breast cancer was 65% at 1-year, 34% at 3-year, and 24% at 5-year follow-up. CONCLUSIONS: Patients with uveal metastasis from breast cancer presented to ophthalmologists with visual symptoms in 93% of cases. However, asymptomatic metastases were commonly detected in the fellow eye. Local ocular tumor control was excellent with current therapies. However, systemic prognosis for all patients, including those who had been treated with different management options, was poor with survival rates of 65% at 1-year and 24% at 5-year follow up. PMID- 12888049 TI - Regional variability in visual field sensitivity during hypercapnia. AB - PURPOSE: Previous investigations have demonstrated a relative vascular autoregulatory inefficiency of the inferior compared to the superior retina in healthy subjects breathing increased CO(2). The purpose of this study was to determine whether the superior and inferior visual field sensitivities of healthy eyes are similarly affected during mild hypercapnia. DESIGN: Experimental study. METHODS: Visual field analysis (Humphrey Field Analyser; SITA standard 24-2 program) was carried out on one randomly selected eye of 22 subjects (mean age, 27.7 +/- 5 years) during normal room air breathing and isoxic hypercapnia. The Student paired t-tests were used to compare the visual field indices mean deviation (MD) and pattern standard deviation (PSD) for each breathing condition. A secondary, sectoral analysis of mean pointwise sensitivity was performed for each condition. In each case a P value of <.01 was considered statistically significant (Bonferroni corrected). RESULTS: Visual field MD was -0.23 +/- 0.95dB during room air breathing and -0.49 +/- 1.04dB during hypercapnia (P =.034). Sectoral pointwise mean sensitivity deteriorated by 0.46dB (P =.006) in the upper visual hemifield during hypercapnia, whereas no significant difference was observed for the lower hemifield (P =.331). CONCLUSIONS: The upper visual hemifield exhibited a significantly greater degree of deterioration in pointwise visual field mean sensitivity compared to the lower hemifield during hypercapnic conditions. This suggests that the upper visual hemifield and hence inferior retina is more susceptible to insult during hypercapnia than the superior retina in healthy individuals. A regional susceptibility of inferior retinal function to altered vascular or metabolic effects may account for the earlier and more frequent inferior nerve fibre damage associated with glaucomatous optic neuropathy. PMID- 12888050 TI - Surgical management of retinal detachment associated with myopic macular hole: anatomic and functional status of the macula. AB - PURPOSE: To evaluate the postoperative status of the macula after vitreous surgery with internal limiting membrane removal for macular hole related retinal detachment in patients with severe myopia. DESIGN: Interventional case series. METHODS: We prospectively examined 10 eyes with retinal detachment associated with a myopic macular hole from 10 consecutive patients, and performed pars plana vitrectomy with internal limiting membrane peeling. Macular buckling was performed in one eye during the initial treatment and in three eyes during subsequent operations. The main outcome measures were the anatomic reattachment rate and the postoperative status of the macular hole. We examined the macular area pre- and postoperatively with slit-lamp biomicroscopy and with a scanning laser ophthalmoscope. Cross-sectional imaging of the macular area was conducted with optical coherence tomography. RESULTS: Successful retinal reattachment was achieved in seven eyes (70%) after the initial surgery and in three eyes (30%) after additional procedures. Visual acuity remained unchanged in two eyes (20%), and improved by two or more logarithmic units of minimum angle of resolution (logMAR) measurement in eight eyes (80%). The macular hole was anatomically closed in only one eye (10%). Postoperative enlargement of the macular hole was observed in seven eyes. CONCLUSIONS: In highly myopic eyes with macular hole related retinal detachment, closure of the macular hole is difficult to attain despite the complete relief of tangential traction by internal limiting membrane peeling. Results indicate the presence of a possible imbalance between the retina and the choroid-sclera complex associated with axial elongation and posterior staphyloma in highly myopic eyes. PMID- 12888051 TI - Validity and reliability of the Cataract TyPE Spec: an instrument for measuring outcomes of cataract extraction. AB - PURPOSE: To design and validate a survey instrument that measures vision-related functional status, which is appropriate for self-administration in a clinical practice and mail survey setting. DESIGN: Observational, validation study in sequential patients. METHODS: A prototype visual function instrument (the Cataract TyPE Spec) was developed based on focus group input and then validated in an outcomes study conducted among 1,823 patients who underwent cataract extraction performed by 22 surgeons at six centers in the United States. Reliability was determined by calculating Cronbach alpha for different types of administration (self-administration at the site of care and mailed survey) and across race and gender. Criterion validity was determined by correlating scores on the TyPE Spec with baseline measures and changes in visual acuity, overall rating of vision, and general quality of life. RESULTS: The instrument was internally valid (Chronbach alpha = 0.94), both on self-administration in the patient care setting and upon mailed survey administration and across patient race and gender. The TyPE Spec score was highly correlated with overall rating of vision (r =.54, P <.0001) and moderately correlated with Snellen acuity (r =.32, P <.0001), rating of vision in the better eye (r =.42, P <.0001), and quality of life, as measured by the physical component summary score of the SF-36 (r =.27, P <.001). Change in TyPE Spec similarly correlated with change in rating of vision overall, change in best-corrected Snellen acuity, and change in rating of vision in the operated eye. CONCLUSIONS: The cataract TyPE Spec instrument was found to be equally valid and internally consistent when administered in the patient care setting and by mail survey. PMID- 12888053 TI - Case-control study of endophthalmitis after cataract surgery comparing scleral tunnel and clear corneal wounds. AB - PURPOSE: To study the possible association between clear corneal incision with or without placement of a suture during cataract extraction and postoperative endophthalmitis. DESIGN: Retrospective, comparative, case-controlled study. METHODS: Thirty-eight patients treated for culture-positive, acute post-cataract surgery endophthalmitis, and 371 randomly selected control patients who underwent uncomplicated cataract surgery in the referring community were studied. Incision type and use of suture during cataract surgery of endophthalmitis patients were compared with the controls. The data were analyzed using logistic regression methods. RESULTS: Of the 38 patients with endophthalmitis, 17 patients (45%) had clear corneal incisions and 21 patients (55%) had a scleral tunnel incision. In 371 controls, 76 patients (20%) had clear corneal incisions and 295 patients (80%) had scleral tunnel incisions. Clear corneal incision was associated with a threefold greater risk of endophthalmitis than was scleral tunnel incision (Odds Ratio, 3.36, 95% Confidence Interval = 1.67 to 6.78). The type of incision was significant (chi(2) = 11.53, P =.0007); a clear corneal incision was more frequently associated with endophthalmitis. A subgroup analysis revealed that the presence or absence of a suture was not significant (chi(2) = 1.31, P =.2524). CONCLUSIONS: In this retrospective, case-controlled study, clear corneal incisions were found to be a statistically significant risk factor for acute post cataract surgery endophthalmitis when compared with scleral tunnel incisions. PMID- 12888052 TI - Abnormal protein profiles in tears with dry eye syndrome. AB - PURPOSE: To verify the hypothesis that protein concentrations, such as lactoferrin, epidermal growth factor (EGF), and aquaporin 5 (AQP5), in tears are abnormal in patients with dry eye. DESIGN: Prospective case-control study. METHODS: One hundred three dry eye patients were divided into three groups: dry eye not associated with the Sjogren syndrome (non-SS; n = 71), Sjogren syndrome (SS; n = 23), and Stevens-Johnson syndrome (SJS; n = 9). Sixteen normal control subjects were also checked. The concentrations of lactoerrin, EGF, and AQP5 were measured by enzyme-linked immunosorbent assay. RESULTS: The concentration of lactoferrin was significantly decreased in tears of non-SS (P =.0001), SS (P =.00005), and SJS (P =.0006) patients compared with control subjects. The concentration of EGF was significantly decreased in non-SS (P =.0005), SS (P =.00002), and SJS (P =.0001) patients compared with control subjects. The concentration of AQP5 was significantly increased in tears of only SS patients (P =.01) compared with control subjects and increased in tears of only SS patients compared with non-SS patients (P =.007). CONCLUSIONS: The decrease in both lactoferrin and EGF was found not only in SS patients but also in non-SS patients, indicating that tear components in dry eyes differ in their quantity and quality. Quantification of AQP5 increased only in SS patients, suggesting that AQP5 protein leaks into the tears when acinar cells of the lacrimal gland are damaged by lymphocytic infiltration. PMID- 12888054 TI - Comparison of the clinical expression of retinitis pigmentosa associated with rhodopsin mutations at codon 347 and codon 23. AB - PURPOSE: To examine the difference in expression of retinitis pigmentosa from mutations at codon 23 and codon 347 or rhodopsin; to report a novel mutation in rhodopsin. METHODS: Goldmann perimetry (solid angle of I4e isopter) and electroretinographic amplitudes (square root transform of a response ratio) were analyzed for 24 patients with mutations at codon 347 (15 with Pro347Ala, 2 with Pro347Gln, 6 with Pro347Leu, and 1 with a novel Pro347Cys change) and 41 patients with mutations at codon 23 (6 with Pro23Ala; 35 with Pro23His). RESULTS: When all patients with mutations at codons 347 and 23 were compared, loss of visual fields was significantly worse in patients with codon 347 changes (P =.0003). Only rod responses of the electroretinograms were significantly different between the two groups (P =.048). Specific comparison of Pro347Ala with Pro23Ala using regression analysis demonstrated significant differences in severity between codon 23 and codon 347 patients for b-wave amplitudes of rod (P =.0069), cone (P =.039) and maximum combined response (P =.049). The solid angle of the I4e isopter was also significantly different (P =.025) between the groups after controlling for age. Modeling age by group for Pro347Ala comparison produced an R(2) of.44. CONCLUSION: We reconfirmed that rhodopsin-related retinitis pigmentosa from mutations involving codon 347 produces a more severe phenotype than that involving codon 23. Accurate modeling of disease was shown to be possible by incorporating the effects of a patient's age and specific genotype. Therefore, both of these variables must be considered in prognostic counseling and subject recruitment for future therapeutic trials. PMID- 12888055 TI - The Eger Macular Stressometer: pilot study. AB - PURPOSE: To evaluate the sensitivity of the Eger Macular Stressometer (EMS) for early screening of age-related macular degeneration (AMD) in a clinical practice. We examined the null hypothesis that AMD eyes have EMS recovery times (RTs) that do not differ from eyes with cataract, diabetic retinopathy, or glaucoma. DESIGN: The design of this study was a nonrandomized clinical trial. METHODS: Ninety-two eyes from 92 patients with vision 20/80 or better, age 50 and older, of either gender, and any ethnic origin, were recruited into one of four groups: AMD (30 eyes), normal or mild cataract (30 eyes), diabetic retinopathy (16 eyes), and glaucoma (16 eyes). Recovery times were obtained with the EMS, according to manufacturer's instructions. RESULTS: The mean (SD) [median] RT for the AMD group was 11.8 (7.6) [9] seconds, the normal/cataract group 10.0 (4.3) [9] seconds, the diabetic retinopathy group 8.4 (3.0) [8] seconds, and glaucoma group 8.6 (2.4) [8] seconds. Recovery time did not appear to be related to group (P =.58), age (P =.50), visual acuity (P =.52), or sex (P =.23). CONCLUSIONS: We found EMS RT distributions did not differ between AMD, cataract, diabetic retinopathy, and glaucoma groups. The EMS in its current form is not a sensitive screening tool for AMD. Further testing is needed to examine EMS sensitivity with other macular diseases such as central serous choroidopathy and diabetic macular edema. PMID- 12888056 TI - Prevalence of dry eye syndrome among US women. AB - PURPOSE: Dry eye syndrome (DES) is believed to be one of the most common ocular problems in the United States (US), particularly among older women. However, there are few studies describing the magnitude of the problem in women and how this may vary with demographic characteristics. DESIGN: Cross-sectional prevalence survey. METHODS: STUDY POPULATION: we surveyed 39,876 US women participating in the Women's Health Study about a history of diagnosed DES and dry eye symptoms. MAIN OUTCOME MEASURE: we defined DES as the presence of clinically diagnosed DES or severe symptoms (both dryness and irritation constantly or often). We calculated the age-specific prevalence of DES and adjusted the overall prevalence to the age distribution of women in the US population. We used logistic regression to examine associations between DES and other demographic factors. RESULTS: The prevalence of DES increased with age, from 5.7% among women < 50 years old to 9.8% among women aged > or = 75 years old. The age-adjusted prevalence of DES was 7.8%, or 3.23 million women aged > or = 50 in the US. Compared with Whites, Hispanic (odds ratio [OR] = 1.81, confidence interval [CI] = 1.18-2.80) and Asian (OR = 1.77, CI = 1.17-2.69) women were more likely to report severe symptoms, but not clinically diagnosed DES. There were no significant differences by income (P([trend]) =.78), but more educated women were less likely to have DES (P([trend]) =.03). Women from the South had the highest prevalence of DES, though the magnitude of geographic differences was modest. CONCLUSIONS: Dry eye syndrome leading to a clinical diagnosis or severe symptoms is prevalent, affecting over 3.2 million American women middle-aged and older. Although the condition is more prevalent among older women, it also affects many women in their 40s and 50s. Further research is needed to better understand DES and its impact on public health and quality of life. PMID- 12888057 TI - Separate effects of the microkeratome incision and laser ablation on the eye's wave aberration. AB - PURPOSE: To study the optical changes induced by the microkeratome cut, the subsequent laser ablation, and the biomechanical healing response of the cornea in normal laser in situ keratomileusis (LASIK) eyes. DESIGN: Prospective randomized clinical trial. METHODS: A Hansatome microkeratome was used to cut a corneal flap in one eye (study eye) of 17 normal myopic patients and a subsequent laser ablation was performed 2 months after this initial microkeratome incision. Control eyes received conventional LASIK treatments at the latter time point. The wave aberration of both the study and contralateral control eyes were measured over a 6-mm pupil with a Shack-Hartmann wavefront sensor for all preoperative, postflap cut, and postablation visits. RESULTS: The eye's higher order aberrations had a small, but significant increase (P =.03) of approximately 30% 2 months after cutting a flap. No systematic changes were observed in nearly all Zernike coefficients from their preoperative levels at 2 months postflap cut. A significant difference between the study and control eyes was observed for one trefoil mode, Z(3)(3) (P =.04). CONCLUSIONS: There was a wide variation in the response of individual Zernike modes across patients after cutting a flap. The majority of spherical aberration induced by the LASIK procedure seems to be due to the laser ablation and not the microkeratome cut. In addition, the total and higher order root mean square of wavefront errors were nearly identical for both the study and control eyes 3-months after the laser ablation, indicating that a procedure in which the incision and the ablation are separated in time to better control aberrations does not compromise the outcome of a conventional LASIK treatment. PMID- 12888058 TI - Medical evaluation of patients before ocular surgery. PMID- 12888059 TI - Wave-front analysis: are we missing something? PMID- 12888060 TI - Ocular complications of smallpox vaccination. AB - PURPOSE: To describe the ocular complications of smallpox vaccination and to discuss potential therapeutic options. DESIGN: Review of pertinent medical literature and recent treatment recommendations of the Centers for Disease Control and Prevention. RESULTS: After immunization against smallpox, vaccinia infection of the eyelid, conjunctiva, or ocular surface can result from accidental autoinoculation from a vaccination site before scab formation or from contact with a recently vaccinated individual. While uncommon, corneal involvement can lead to stromal opacification and scarring. Clinical findings of ocular and periocular vaccinia must be differentiated from those produced by other pathogens such as molluscum contagiosum, herpes simplex, varicella zoster, and acanthamoeba infections. Clinical diagnosis can be confirmed by electron microscopy to identify the presence of orthopoxvirus, as well as by virologic culture, polymerase chain reaction, and/or restriction endonuclease analysis of viral isolates. CONCLUSIONS: While the majority of ocular complications of smallpox vaccination in immunocompetent patients are self-limiting, selective cases may require treatment with trifluridine drops, topical corticosteroids and vaccinia immune globulin (VIG). Vaccinia virus does not appear to be sensitive to acyclovir. Specific treatment recommendations are outlined for the spectrum of ocular manifestations. PMID- 12888061 TI - Persistent subretinal indocyanine green induces retinal pigment epithelium atrophy. AB - PURPOSE: To describe a case of a patient with macular hole with subretinal indocyanine green (ICG) during vitrectomy. DESIGN: Interventional case report. METHODS: A 66-year-old woman with macular hole underwent a vitrectomy with ICG. RESULTS: After application of ICG into the vitreous, ICG was introduced in the subretinal space. Indocyanine green was found to be present for more than 6 months. Retinal pigment epithelium atrophy appeared at the site of the lesion. CONCLUSIONS: Although ICG may be a useful tool for distinguishing the internal limiting membrane and other tissues careful application is required to prevent side effects. PMID- 12888062 TI - Accessory lateral rectus muscle in a patient with congenital third-nerve palsy. AB - PURPOSE: To report a case of accessory lateral rectus muscle in a patient with congenital third-nerve palsy. DESIGN: Observational case report. METHODS: An 18 year-old boy with left exodeviation, ptosis, pupil dilation, and limited adduction, supraduction, and infraduction of his left eye. Left lateral rectus muscle recession and medial rectus muscle resection were done. An orbital computed tomographic (CT) scan was obtained. RESULT: Intraoperatively, an accessory muscle was found under the lateral rectus muscle. Postoperatively, the orbital CT scan showed accessory lateral rectus muscle located in the medial side of the lateral rectus muscle. CONCLUSION: Accessory lateral rectus muscle was demonstrated in a patient with congenital third-nerve palsy using lateral rectus muscle surgery and an orbital CT scan. PMID- 12888063 TI - Chryseobacterium meningosepticum keratitis. AB - PURPOSE: To describe a case of a corneal ulcer caused by Chryseobacterium meningosepticum in an eye without significant comorbidities. DESIGN: Interventional case report. METHODS: Retrospective case review. RESULTS: A 48 year-old man presented with a corneal ulcer in his right eye and was treated with ofloxacin and fortified bacitracin ophthalmic solution. Cultures grew C. meningosepticum, and the infection resolved. Two previous case reports in the literature have described ocular infections with this bacterium, both in postoperative patients with significant ocular comorbidities. CONCLUSIONS: This is the first report of ocular infection with C. meningosepticum in an eye with no significant comorbidities. PMID- 12888064 TI - Autoimmune optic neuropathy with anticardiolipin antibody mimicking multiple sclerosis in a child. AB - PURPOSE: To demonstrate that autoimmune optic neuropathy (AON) may occur in the pediatric population. DESIGN: Interventional case report. METHODS: A 4-year-old developed four episodes of bilateral optic neuritis with mild concurrent weakness, ataxia, or dizziness; one episode of unilateral optic neuritis; and one episode of weakness over a period of 4 years. RESULTS: Autoimmune optic neuropathy was diagnosed because of the presence of anticardiolipin antibody and an abnormal skin biopsy with thrombin and immunoreactant deposition. She was treated with corticosteroids, aspirin, and gammaglobulin. This diminished the frequency and intensity of her attacks. CONCLUSION: This case represents the diagnosis and treatment of AON in a child. PMID- 12888065 TI - Intraocular pressure in patients with human immunodeficiency virus and treated with highly active antiretroviral therapy. AB - PURPOSE: To compare the intraocular pressure (IOP) of patients infected with human immunodeficiency virus (HIV) and treated with highly active antiretroviral therapy (HAART) to a control group and to determine whether HAART reverses the previously reported reduction in IOP associated with HIV infection. DESIGN: Retrospective case control study. METHODS: Review of patients with HIV infection on HAART compared with a matched control group. RESULTS: Among 64 patients and 56 control subjects, no significant difference in IOP was found (P =.520), nor was correlation noted between IOP and either CD4+ T-lymphocyte counts, or viral load. CONCLUSION: When HIV infection is treated with HAART, IOP is not significantly different from that of control subjects. PMID- 12888066 TI - Alterations in retinal nerve fiber layer thickness following indirect traumatic optic neuropathy detected by nerve fiber analyzer, GDx-N. AB - PURPOSE: To evaluate changes in the retinal nerve fiber layer following traumatic optic neuropathy. DESIGN: Observational case report. METHODS: A patient presented with visual loss after an accident. Scanning laser polarimetry was performed. RESULTS: Thickness of the retinal nerve fiber layer increased immediately after the trauma but then progressively decreased. Severe loss was observed at day 90 and then ceased. Enlargement of the optic disk cup was also observed. CONCLUSIONS: This is the first report documenting early transient increase followed by progressive loss of the retinal nerve fiber layer in traumatic optic neuropathy. PMID- 12888067 TI - Slit-lamp visualization of aqueous flow through an inferior iridectomy. AB - PURPOSE: To demonstrate how an inferior iridectomy prevents a pupillary block in aphakic patients with a silicone oil tamponade. DESIGN: Observational case series. METHODS: Photographs of two patients who had undergone vitrectomy, lensectomy, membrane peeling, silicone oil tamponade, and inferior iridectomy for a complex retinal detachment were evaluated. RESULTS: In the thin layer of erythrocytes covering the endothelium, we observed an arc clear of erythrocytes anterior to the inferior iridectomy. CONCLUSIONS: With silicone oil blocking the pupil, aqueous humor was channeled through the inferior iridectomy with sufficient force to sweep the endothelium anterior to the iridectomy clean of erythrocytes, thus visualizing the mechanism that prevents a pupillary block glaucoma. PMID- 12888068 TI - Intraocular dispersion of perfluorocarbon liquids in silicone oil. AB - PURPOSE: To report one case of intraocular emulsification of perfluorocarbon liquids (PFCLs) and silicone oil after vitrectomy surgery. DESIGN: Interventional case report. METHODS: Retrospective review of the clinic, and surgical record of a 39-year old female referred for persistent retinal detachment after vitrectomy surgery; PFCLs and silicone oil were left in the eye at the end of the surgery. RESULTS: There was intraocular emulsification of PFCLs into silicone oil. Electron microscope examination of the silicone oil surgically extracted from the eye demonstrated emulsification of small PFCL bubbles into the silicone oil. CONCLUSION: Perfluorocarbon liquids are surgical tools. They should be completely removed from the eye at the end of the surgery; PFCLs and silicone oil tend to emulsify together, originating an opaque fluid, which contains both silicone oil and dispersed PFCL bubbles. PMID- 12888069 TI - Submacular surgery for choroidal neovascularization secondary to optic nerve drusen. AB - PURPOSE: To report a case of bilateral choroidal neovascularization (CNV) secondary to optic nerve drusen (OND). DESIGN: Interventional case report. METHODS: A 9-year-old girl presented with peripapillary CNV of the right eye and peripapillary and subfoveal CNV of the left eye secondary to OND. The CNV was excised with submacular surgery in the left eye. RESULTS: Visual acuity increased from 0.05 to 0.3 in the left eye after submacular surgery and did not change from 0.3 in the right eye during 6-month follow-up. Partial retinal pigment epithelial atrophy, but no recurrence of CNV, was observed after CNV excision. CONCLUSION: Subfoveal CNV is an uncommon complication of OND and surgical excision may be useful in selected cases. PMID- 12888070 TI - Toxoplasmosis transmitted to a newborn from the mother infected 20 years earlier. AB - PURPOSE: To present a case of congenital toxoplasmosis in a newborn whose mother had a 20-year history of a chorioretinal macular scar and positive serology for toxoplasmosis. DESIGN/METHODS: Case report. SETTING/RESULTS: A 38-year-old woman who had been treated for ocular toxoplasmosis 20 years earlier delivered a newborn who presented with a focal necrotizing retinochoroiditis characteristic of toxoplasmosis, as well as positive immunoglobulin (Ig) G and M serology for toxoplasmosis. The workup was negative for other entities. CONCLUSION: This case suggests that women with old retinal scars due to toxoplasmosis and long-standing IgG antibodies to toxoplasmosis are also at risk of transmitting this disease to the fetus. PMID- 12888071 TI - Photodynamic therapy for juxtafoveal choroidal neovascularization in myopic eyes. AB - PURPOSE: To report the results of photodynamic therapy in myopic patients with juxtafoveal choroidal neovascularization (CNV). DESIGN: Interventional case series. METHODS: Three consecutive myopic patients with juxtafoveal CNV were treated by photodynamic therapy with the same protocol as the one used to treat subfoveal CNV. Visual acuity was measured on Early Treatment Diabetic Retinopathy Study (ETDRS) charts by an independent observer. Fluorescein angiography was performed 6 and 12 weeks after photodynamic therapy and then every 12 weeks. RESULTS: In all three patients, one treatment of photodynamic therapy using verteporfin resulted in the involution of the CNV and improvement in best corrected visual acuity from 20/50 or 20/40 to 20/25 or more. No retreatment was required during follow-up, which lasted for 12 to 24 months (mean = 17 months). CONCLUSION: Photodynamic therapy may be an effective treatment for juxtafoveal CNV in myopic eyes. PMID- 12888072 TI - Presumed minor salivary gland secretion in a patient with a history of oral mucous membrane graft. AB - PURPOSE: To describe presumed minor salivary gland secretion in a patient with a history of oral mucous membrane graft in conjunctival plastic surgery. DESIGN: Interventional case report. METHODS: A 74-year-old woman who had had an oral mucous membrane graft in the left eye complained of mucous discharge. Slit-lamp examination revealed a yellowish-red mass lesion in the superior bulbar conjunctiva. RESULTS: The lesion was excised. Histopathologic examination of the excised tissue revealed lobules and a duct of minor salivary glands. After removal of the mass, mucous discharge decreased. CONCLUSION: Minor salivary glands within a mucous membrane graft may cause mucous discharge in the eye. PMID- 12888073 TI - Intumescent cataract after topical mitomycin-C for conjunctival malignant melanoma. AB - PURPOSE: To present the clinical and histologic findings of a patient in whom intumescent cataract developed after successful topical mitomycin-C (MMC) chemotherapy for conjunctival melanoma originating from primary acquired conjunctival melanosis (PAM) with atypia. DESIGN: Observational case report; follow-up at 30 months. METHODS: In a patient with PAM and subsequent conjunctival melanoma that was successfully treated with topical MMC chemotherapy an intumescent cataract developed after two cycles of 0.04% MMC, each for 1 month. RESULTS: After MMC chemotherapy pigmentation of the acquired melanosis vanished almost completely. Histopathology of the check-up specimen revealed local tumor control. Six weeks after the completion of the second cycle, an intumescent cataract developed. Cataract surgery was performed uneventfully. The patient was followed up for 30 months. CONCLUSIONS: In selected cases, topical MMC chemotherapy is effective for treating conjunctival melanoma. Although severe complications are rare and usually transient, development of cataract may be observed. A prospective study should be initiated. PMID- 12888074 TI - Corneal iron ring after conductive keratoplasty. AB - PURPOSE: To report formation of corneal iron ring deposits after conductive keratoplasty. DESIGN: Observational case report. METHODS: Case report. RESULTS: A 54-year-old woman underwent conductive keratoplasty for hyperopia. One year after conductive keratoplasty, iron ring pattern pigmentation was detected at the corneal epithelium of both eyes. CONCLUSIONS: This is the first report of the appearance of corneal iron ring deposits following conductive keratoplasty treatment in a patient. It is suggested that alterations in tear film stability, resulting from conductive keratoplasty-induced changes in corneal curvature, constitute the contributory factor for these deposits. PMID- 12888075 TI - A case of primary B-cell type non-Hodgkin lymphoma originating in the iris. AB - PURPOSE: To report a case of primary malignant B-cell type non-Hodgkin lymphoma originating in the iris. DESIGN: Interventional case report. METHODS: An 83-year old woman presented with anterior uveitis resulting from primary malignant lymphoma in the iris. Ultrasound biomicroscopy and indocyanine green angiography using a scanning laser ophthalmoscope showed abnormalities in the iris. Diagnostic biopsy of the iris revealed B-cell type non-Hodgkin lymphoma. RESULT: The patient was treated with radiotherapy, and the tumorous lesion resolved. CONCLUSION: Primary lymphoma localized in the iris only is rare. In this case, diagnostic biopsy and radiotherapy of the iris lymphoma provided good results. PMID- 12888076 TI - Subluxation of transscleral sutured posterior chamber intraocular lens (TSIOL). AB - PURPOSE: To evaluate factors related to late-onset transscleral sutured posterior chamber intraocular lens (TSIOL) subluxation. METHODS: Retrospective observational case series. Analysis of subluxated TSIOLs in seven eyes from seven patients treated between May 1999 and May 2001. RESULTS: Mean age at the time of TSIOL surgery was 33 +/- 6 years. Mean time from TSIOL surgery to its subluxation was 78 +/- 19 months. Initial diagnoses requiring TSIOL surgeries were previous history of trauma and Marfan syndrome. Subluxation of TSIOLs was associated with blunt trauma in three eyes, whereas the other four eyes experienced spontaneous lens dislocation. CONCLUSION: Subluxation of TSIOL is not uncommon in younger patients with history of trauma or Marfan syndrome. PMID- 12888077 TI - Intravitreal triamcinolone acetonide for pseudophakic cystoid macular edema. AB - PURPOSE: To report the clinical outcome of patients undergoing intravitreal injection of triamcinolone acetonide as treatment of long-standing cystoid macular edema after phacoemulsification. DESIGN: Prospective clinical interventional cases series studies. METHODS: The study included five patients suffering from cystoid macular edema after cataract surgery. They received an intravitreal injection of 25-mg crystalline triamcinolone acetonide transconjunctivally with topical anesthesia. RESULTS: In the follow-up period of 6.6 +/- 4.1 months, visual acuity increased from 0.26 +/- 0.13 to a mean maximal visual acuity of 0.60 +/- 0.19. For all patients, visual acuity improved during the follow-up by at least 0.20. Two (40%) patients developed intraocular pressure values higher than 21 mm Hg, which could be controlled by topical antiglaucomatous treatment. CONCLUSIONS: Intravitreal triamcinolone acetonide may be a therapeutic option for long-standing cystoid macular edema after cataract surgery. PMID- 12888078 TI - Cytomegalovirus retinitis and FK 506. AB - PURPOSE: To report a case of cytomegalovirus (CMV) retinitis in a patient immunosuppressed with FK 506. DESIGN: Interventional case report. METHODS: Retrospective institutional study. One patient, diagnosed with bilateral CMV retinitis while on immunosuppressive treatment with FK 506, received systemic ganciclovir and intravitreal, slow-release ganciclovir implant in one eye, combined with reduction in FK 506 dose. Main outcome measures were visual acuity and regression of CMV retinitis. RESULTS: Visual acuity at presentation was 20/20 in the right eye and 20/50 in the left eye. During the 6 months of follow-up, there was no change in visual acuity in either eye. The areas of CMV retinitis demonstrated progressive regression, leaving diffuse retinal atrophy. CONCLUSIONS: Cytomegalovirus retinitis can be a sight-threatening complication of immunosuppressive treatment with FK 506. Ganciclovir treatment together with a reduction in FK 506 dose was effective in preserving vision. PMID- 12888079 TI - Indocyanine green for successful repair of a long-standing macular hole. AB - PURPOSE: To report on the successful closure of a persistent chronic macular hole using indocyanine green (ICG) during the second surgical approach. DESIGN: Interventional case report. METHODS: A patient with a long standing macular hole (18 years) underwent pars plana vitrectomy with peeling of the internal limiting membrane (ILM). While no dye was used during the first operation, ICG (0.05%, Pulsion) was applied during the second procedure. The light source was turned off immediately after the ICG application until the dye was washed out after a period of 15 seconds. Tissue harvested during the second surgical approach was prepared for electron microscopy. RESULTS: After the failure of the first operation, areas of incomplete ILM peeling could be stained using ICG and a thorough ILM removal was achieved. Postoperatively, the macular hole was closed, as confirmed by optical coherence tomography. Visual acuity increased from 20/200 to 20/50. No visual field defects were seen postoperatively. There were only few cellular elements adherent to the retinal surface of the ILM as seen during electron microscopy. CONCLUSION: In cases where tractional forces due to incomplete ILM removal are considered responsible for the persistence of a macular hole, ICG is useful to identify areas of unpeeled ILM. Under certain safety precautions no adverse effect on functional outcome was seen. PMID- 12888081 TI - Six-month randomized clinical trial comparing intraocular pressure-lowering efficacy of bimatoprost and latanoprost in patients with ocular hypertension or glaucoma. PMID- 12888082 TI - Risk factors for intraoperative epithelial defect in laser in situ keratomileusis. PMID- 12888084 TI - Iridociliary apposition in plateau iris syndrome persists after cataract extraction. PMID- 12888086 TI - Iatrogenic punctate chorioretinopathy after internal limiting membrane peeling. PMID- 12888089 TI - Foveal hypoplasia demonstrated in vivo with optical coherence tomography. PMID- 12888090 TI - Association of disk size and glaucomatous cupping. PMID- 12888105 TI - Observations at the interface of mutation research and regulatory policy. AB - Between 1970 and 1975 developments in environmental mutagenesis proceeded with amazing speed. These developments were both structural and conceptual in nature. A new infrastructure was built and new concepts about how best to protect consumers from exposures to mutagens emerged. The internal dynamics within the Food and Drug Administration played an important role and is discussed with regard to modifications in testing protocols as well as changes in the overall approach used to protect consumers. It is clear that this exciting period in the early days of environmental mutagenesis has provided a base for growth and development of the field and continues to affect and guide future developments. PMID- 12888106 TI - Molecular epidemiology of sporadic breast cancer. The role of polymorphic genes involved in oestrogen biosynthesis and metabolism. AB - The major known risk factors for female breast cancer are associated with prolonged exposure to increased levels of oestrogen. The predominant theory relates to effects of oestrogen on cell growth. Enhanced cell proliferation, induced either by endogenous or exogenous oestrogens, increases the number of cell divisions and thereby the possibility for mutation. However, current evidence also supports a role for oxidative metabolites, in particular catechol oestrogens, in the initiation of breast cancer. As observed in drug and chemical metabolism, there is considerable interindividual variability (polymorphism) in the conjugation pathways of both oestrogen and catechol oestrogens. These person to-person differences, which are attributed to polymorphisms in the genes encoding for the respective enzymes, might define subpopulations of women with higher lifetime exposure to hormone-dependent growth promotion, or to cellular damage from particular oestrogens and/or oestrogen metabolites. Such variation could explain a portion of the cancer susceptibility associated with reproductive effects and hormone exposure. In this paper the potential role of polymorphic genes encoding for enzymes involved in oestrogen biosynthesis (CYP17, CYP19, and 17beta-HSD) and conversion of the oestrogen metabolites and their by-products (COMT, CYP1A1, CYP1B1, GSTM1, GSTM3, GSTP1, GSTT1 and MnSOD) in modulating individual susceptibility to breast cancer are reviewed. Although some of these low-penetrance genes appeared as good candidates for risk factors in the etiology of sporadic breast cancer, better designed and considerably larger studies than the majority of the studies conducted so far are evidently needed before any firm conclusions can be drawn. PMID- 12888107 TI - Use of the single cell gel electrophoresis/comet assay for detecting DNA damage in aquatic (marine and freshwater) animals. AB - The comet assay is a rapid, sensitive and inexpensive method for measuring DNA strand breaks. The comet assay has advantages over other DNA damage methods, such as sister chromatid exchange, alkali elution and micronucleus assay, because of its high sensitivity and that DNA strand breaks are determined in individual cells. This review describes a number of studies that used the comet assay to determine DNA strand breaks in aquatic animals exposed to genotoxicants both in vitro and in vivo, including assessment of DNA damage in aquatic animals collected from contaminated sites. One difficulty of using the comet assay in environmental work is that of comparing results from studies that used different methods, such as empirical scoring or comet tail lengths. There seems to be a consensus in more recent studies to use both the intensity of the tail and the length of the tail, i.e. DNA tail moment, percentage of DNA in the tail. The comet assay has been used to assess DNA repair and apoptosis in aquatic animals and modifications of the comet assay have allowed the detection of specific DNA lesions. There have been some recent studies to link DNA strand breaks in aquatic animals to effects on the immune system, reproduction, growth, and population dynamics. Further work is required before the comet assay can be used as a standard bio-indicator in aquatic environments, including standardization of methods (such as ASTM method E2186-02a) and measurements. PMID- 12888108 TI - Theoretical analysis of mutation hotspots and their DNA sequence context specificity. AB - Mutation frequencies vary significantly along nucleotide sequences such that mutations often concentrate at certain positions called hotspots. Mutation hotspots in DNA reflect intrinsic properties of the mutation process, such as sequence specificity, that manifests itself at the level of interaction between mutagens, DNA, and the action of the repair and replication machineries. The hotspots might also reflect structural and functional features of the respective DNA sequences. When mutations in a gene are identified using a particular experimental system, resulting hotspots could reflect the properties of the gene product and the mutant selection scheme. Analysis of the nucleotide sequence context of hotspots can provide information on the molecular mechanisms of mutagenesis. However, the determinants of mutation frequency and specificity are complex, and there are many analytical methods for their study. Here we review computational approaches for analyzing mutation spectra (distribution of mutations along the target genes) that include many mutable (detectable) positions. The following methods are reviewed: derivation of a consensus sequence, application of regression approaches to correlate nucleotide sequence features with mutation frequency, mutation hotspot prediction, analysis of oligonucleotide composition of regions containing mutations, pairwise comparison of mutation spectra, analysis of multiple spectra, and analysis of "context-free" characteristics. The advantages and pitfalls of these methods are discussed and illustrated by examples from the literature. The most reliable analyses were obtained when several methods were combined and information from theoretical analysis and experimental observations was considered simultaneously. Simple, robust approaches should be used with small samples of mutations, whereas combinations of simple and complex approaches may be required for large samples. We discuss several well-documented studies where analysis of mutation spectra has substantially contributed to the current understanding of molecular mechanisms of mutagenesis. The nucleotide sequence context of mutational hotspots is a fingerprint of interactions between DNA and DNA repair, replication, and modification enzymes, and the analysis of hotspot context provides evidence of such interactions. PMID- 12888109 TI - Current concepts in psoriasis and its treatment. AB - Many advances have been made in our understanding of the biology of psoriasis over the past 20 years. We are close to knowing the genetic determinants of psoriasis. There is now greater insight into the immunological mechanisms that produce the phenotype, and the possibility of intervening selectively to antagonise some of these mechanisms is becoming a reality. Nevertheless, psoriasis remains an enigmatic disease, and much of the suffering it produces is not adequately addressed. The sheer number of treatments that are used therapeutically suggests a lack of efficacy and/or toxicity of many of these approaches. In this review, we aim to briefly describe the biology of psoriasis, document the key features of treatments that are available or under development, and explain how these treatments can be used effectively to manage this chronic relapsing disease. PMID- 12888110 TI - Pharmacological aspects of anticancer drug-induced emesis with emphasis on serotonin release and vagal nerve activity. AB - Cytotoxic drug-induced nausea and vomiting are the side effects most feared by cancer patients. Emesis is an instinctive defense reaction caused by the somatoautonomic nerve reflex, which is integrated in the medulla oblongata. Emesis caused by cytotoxic drugs such as cisplatin is associated with an increase in the concentration of 5-hydroxytryptamine (5-HT) in the intestine and the brainstem. It is proposed that cytotoxic drugs evoke 5-HT release from the enterochromaffin (EC) cells in the intestinal mucosa and that the released 5-HT stimulates the 5-HT receptors on the adjacent vagal afferent nerves. The depolarization of the vagal afferent nerves stimulates the vomiting center in the brainstem and eventually induces a vomiting reflex. 5-HT released from EC cells seems to mediate the cisplatin-induced emesis sensitive to 5-HT(3) receptor antagonists. The release of 5-HT from the EC cells, however, is regulated by polymodal mechanisms on autoreceptors or heteroreceptors. The precise role of 5 HT on the occurrence of vomiting has not been fully elucidated. The present review aims to describe the role of 5-HT in anticancer drug-induced emesis from the viewpoint of 5-HT release and afferent vagus nerve activity. Various methods for predicting emesis are also evaluated. PMID- 12888111 TI - Catalytic topoisomerase II inhibitors in cancer therapy. AB - The nuclear enzyme DNA topoisomerase II is a major target for antineoplastic agents. All topoisomerase II-directed agents are able to interfere with at least one step of the catalytic cycle. Agents able to stabilize the covalent DNA topoisomerase II complex (also known as the cleavable complex) are traditionally called topoisomerase II poisons, while agents acting on any of the other steps in the catalytic cycle are called catalytic inhibitors. Thus, catalytic topoisomerase II inhibitors are a heterogeneous group of compounds that might interfere with the binding between DNA and topoisomerase II (aclarubicin and suramin), stabilize noncovalent DNA topoisomerase II complexes (merbarone, ICRF 187, and structurally related bisdioxopiperazine derivatives), or inhibit ATP binding (novobiocin). Some, such as fostriecin, may also have alternative biological targets. Whereas topoisomerase II poisons are used solely for their antitumor activities, catalytic inhibitors are utilized for a variety of reasons, including their activity as antineoplastic agents (aclarubicin and MST-16), cardioprotectors (ICRF-187), or modulators in order to increase the efficacy of other agents (suramin and novobiocin). In this review, the mechanism and biological activity of different catalytic inhibitors is described, with emphasis on therapeutically used compounds. We will then discuss future development and applications of this interesting class of compounds. PMID- 12888112 TI - Model systems in drug discovery: chemical genetics meets genomics. AB - Animal model systems are an intricate part of the discovery and development of new medicines. The sequencing of not only the human genome but also those of the various pathogenic bacteria, the nematode Caenorhabditis elegans, the fruitfly Drosophila, and the mouse has enabled the discovery of new drug targets to push forward at an unprecedented pace. The knowledge and tools in these "model" systems are allowing researchers to carry out experiments more efficiently and are uncovering previously hidden biological connections. While the history of bacteria, yeast, and mice in drug discovery are long, their roles are ever evolving. In contrast, the history of Drosophila and C. elegans at pharmaceutical companies is short. We will briefly review the historic role of each model organism in drug discovery and then update the readers as to the abilities and liabilities of each model within the context of drug development. PMID- 12888113 TI - The pathophysiology of falciparum malaria. AB - Falciparum malaria is a complex disease with no simple explanation, affecting organs where the parasite is rare as well as those organs where it is more common. We continue to argue that it can best be understood in terms of excessive stimulation of normally useful pathways mediated by inflammatory cytokines, the prototype being tumor necrosis factor (TNF). These pathways involve downstream mediators, such as nitric oxide (NO) that the host normally uses to control parasites, but which, when uncontrolled, have bioenergetic failure of patient tissues as their predictable end point. Falciparum malaria is no different from many other infectious diseases that are clinically confused with it. The sequestration of parasitized red blood cells, prominent in some tissues but absent in others with equal functional loss, exacerbates, but does not change, these overriding principles. Recent opportunities to stain a wide range of tissues from African pediatric cases of falciparum malaria and sepsis for the inducible NO synthase (iNOS) and migration inhibitory factor (MIF) have strengthened these arguments considerably. The recent demonstration of bioenergetic failure in tissue removed from sepsis patients being able to predict a fatal outcome fulfils a prediction of these principles, and it is plausible that this will be demonstrable in severe falciparum malaria. Understanding the disease caused by falciparum malaria at a molecular level requires an appreciation of the universality of poly(ADP-ribose) polymerase-1 (PARP-1) and Na(+)/K(+)-ATPase and the protean effects of activation by inflammation of the former that include inactivation of the latter. PMID- 12888114 TI - Reducing GABA receptors. AB - A number of important drugs act on GABA(A) receptors, pentameric GABA-gated chloride channels assembled from among 19 known subunits. In trying to discover the roles in the brain of the subunits and their combinations, with the goal of developing more selective drugs, one tool has been to reduce expression of the subunits and examine the functional consequences. After briefly examining the properties of GABA(A) receptors, this review surveys the means available for receptor subunit reduction, and some of the observations to which their application has led. The methods discussed include radiation-induced deletion, gene knockout, knock-in mutations, antisense, ribozymes, RNA interference, dominant negative constructs, and transcriptional regulation, e.g., via decoy oligonucleotides. PMID- 12888115 TI - Blockage of transcription as a trigger for p53 accumulation by 2 acetylaminofluorene DNA-adducts. AB - The hepatocarcinogen 2-acetylaminofluorene is one of the most studied experimental carcinogens. We have shown previously that normal rat hepatocytes accumulate the tumour suppressor p53 after exposure to this compound while preneoplastic rat hepatocytes do not. We suggested that the lack of p53 response may confer a growth advantage on preneoplastic hepatocytes and may be an important factor in hepatic tumor promotion by 2-acetylaminofluorene and other genotoxic compounds. Inhibition of RNA polymerase II driven transcription by DNA lesions may constitute one of the mechanisms leading to accumulation of the tumour suppressor p53. We have investigated the accumulation of p53 by structurally different DNA lesions of 2-acetylaminofluorene for which the rate of nucleotide excision repair (NER) and inhibition of transcription are known. Experiments were performed with NER proficient human fibroblasts as well as repair deficient xeroderma pigmentosum group A (XPA) cells, XPC cells [only transcription coupled repair (TCR)] and Cockayne syndrome (CS)B cells [only global genome repair (GGR)]. The cells were exposed to N-acetoxy acetylaminofluorene (NAAAF) in the presence or absence of paraoxon inducing dG-C8 AAF or dG-C8-AF adducts respectively. Both treatments led to accumulation of p53 in all cells. However, dG-C8-AAF adducts produced greater p53 induction than dG C8-AF adducts. The percentage p53-positive cells was highest and the threshold for p53 accumulation was lowest in XPA and CSB cells. Our results further demonstrate that both the potency of a lesion to inhibit transcription as well as the restoration of RNA synthesis determines the magnitude of p53 induction. PMID- 12888116 TI - The efficacy of trovafloxacin versus ceftriaxone in the treatment of experimental brain abscess/cerebritis in the rat. AB - Current estimates of the mortality associated with brain abscesses range from 0 24%, with neurological sequellae in 30-55% of survivors. Although the incidence of brain abscess appears to be increasing, likely due to an increase in the population of immunosuppressed patients, the condition is still sufficiently uncommon to make human clinical trials of therapy problematic. An animal model to study the efficacy of new treatment regimens, specifically, new antimicrobial agents is therefore necessary. This study uses a well-defined experimental paradigm as an inexpensive method of inducing and studying the efficacy of antibiotics in brain abscess. The rat model of brain abscess/cerebritis developed at this institution was used to determine the relative efficacy of trovafloxacin as compared to ceftriaxone in animals infected with Staphylococcus aureus. S. aureus ( approximately 10(5) CFU in 1 microliter) was injected with a Hamilton syringe, very slowly, over the course of 70 minutes after a two mm burr hole was created with a spherical carbide drill just posterior to the coronal suture and four mm lateral to the midline. Eighteen hours later treatment was begun; every 8 hours the rats were dosed with subcutaneous ceftriaxone (n = 10), trovafloxacin (n = 11) or 0.9% sterile pyogen-free saline (n = 10). After four days of treatment the brains were removed and sectioned with a scalpel. The entire injected hemisphere was homogenized and quantitative cultures performed. The mean +/- SEM log(10) colony forming units/ml S. aureus recovered from homogenized brain were as follows: controls 6.10 +/- 0.28; ceftriaxone 3.43 +/- 0.33; trovafloxacin 3.65 +/- 0.3. There was no significant difference in bacterial clearance between ceftriaxone versus trovafloxacin (p = 0.39). Trovafloxacin or other quinolones may provide a viable alternative to intravenous antibiotics in patients with brain abscess/cerebritis. PMID- 12888117 TI - Effects of cysteine on the pharmacokinetics of intravenous clarithromycin in rats with protein-calorie malnutrition. AB - Effects of cysteine on the pharmacokinetics of clarithromycin were investigated after intravenous administration of the drug at a dose of 20 mg/kg to control rats (4-week fed on 23% casein diet) and rats with PCM (protein-calorie malnutrition, 4-week fed on 5% casein diet) and PCMC (PCM treated with 250 mg/kg for oral cysteine twice daily during the fourth week). Clarithromycin has been reported to be metabolized via hepatic microsomal cytochrome P450 (CYP) 3A4 to 14 hydroxyclarithromycin (primary metabolite of clarithromycin) in human subjects. It has also been reported that in rats with PCM, CYP3A23 level decreased to 40 50% of control level, but decreased CYP3A23 level in rats with PCM completely returned to control level by oral cysteine supplementation (rats with PCMC). Human CYP3A4 and rat CYP3A23 proteins have 73% homology. In rats with PCM, the area under the plasma concentration-time curve from time zero to time infinity, AUC (567, 853 and 558 microg min/ml for control rats and rats with PCM and PCMC, respectively) and percentage of clarithromycin remaining after incubation with liver homogenate (69.6, 83.9 and 71.7%) were significantly greater than those in control rats and rats with PCMC. Moreover, in rats with PCM, the total body clearance, CL (35.3, 23.4 and 35.8 ml/min/kg), nonrenal clearance, CL(NR) (21.3, 15.2 and 24.1 ml/min/kg) and maximum velocity for the disappearance of clarithromycin after incubation with hepatic microsomal fraction, V(max) (351, 211 and 372 pmol/min/mg protein) were significantly slower than those in control rats and rats with PCMC. However, above mentioned each parameter was not significantly different between control rats and rats with PCMC. The above data suggested that metabolism of clarithromycin decreased significantly in rats with PCM as compared to control due to significantly decreased level of CYP3A23 in the rats. By cysteine supplementation (rats with PCMC), some pharmacokinetic parameters of clarithromycin (AUC, CL, CL(NR) and V(max)) were restored fully to control levels because CYP3A23 level was completely returned to control level in rats with PCMC. PMID- 12888118 TI - Quantitative assessment of DNA fragmentation and beta-amyloid deposition in insular cortex and midfrontal gyrus from patients with Alzheimer's disease. AB - It has been suggested that the neurodegeneration that occurs with Alzheimer's disease (AD) may result from apoptosis, a process of programmed cell death. Neuronal injury, induced by abnormal aggregates of beta-amyloid peptide, has been identified as an apoptotic trigger. In the present study, brain tissue samples were obtained from the insular cortex (INS) and midfrontal gyrus (MFG) of Alzheimer subjects and age-matched, nondemented controls. Tissue sections from all samples were alternately stained by an in situ TUNEL assay to identify 3' termini DNA strand breaks characteristic of apoptosis or immunohistochemically for beta-amyloid deposition in senile plaques. The incidence of DNA fragmentation detected in pyramidal neurons was relatively infrequent overall, but was significantly higher in AD compared to controls. AD subjects consistently exhibited a dense accumulation of plaques, with a twofold greater concentration in MFG as INS. There was no significant difference in pyramidal cell number regardless of subject or brain region. Taken together, our results indicate that the TUNEL assay may be revealing cell damage rather than cell loss. Our finding of a moderate correlation between the incidence of TUNEL-positive cells and plaque density implicates beta-amyloid as one of multiple factors provoking cell injury in AD. A notable contribution of this study is the identification of distinctive neuropathologies co-occurring in two brain regions interconnected with each other and with limbic and cortical areas typically damaged during AD. PMID- 12888119 TI - Modulation of intracellular Ca(2+) concentration by tedisamil, a class III antiarrhythmic agent, in isolated heart preparation. AB - New class III antiarrhythmic/defibrillating compound tedisamil was shown to facilitate termination of atrial and ventricular fibrillation in experimental as well as clinical conditions. However, class III-related inhibition of K(+) current associated with prolongation of repolarization can not solely explain its defibrillating ability. Following recent findings it was hypothesized that defibrillating effect of tedisamil is likely due to its sympathomimetic feature linked with modulation of intracellular calcium. Results of this study obtained in isolated heart preparation showed that elevated intracellular Ca(2+) free concentration was decreased by administration of tedisamil in concentration that did not induce Q-T interval prolongation. Due to species differences the effective concentration was in rat 10(-7) M, while in guinea pig 10(-5) M. On the contrary, further dramatic increase of elevated Ca(2+) was detected upon administration of tedisamil in concentration that markedly prolonged Q-T interval (10(-5) M in rat). It is concluded that defibrillating ability of tedisamil is most likely associated with attenuation of abnormal and harmful intracellular Ca(2+) elevation (that is highly arrhythmogenic) than with prolongation of APD or Q-T interval. PMID- 12888120 TI - Effect of the NAD(P)H oxidase inhibitor, apocynin, on peripheral nerve perfusion and function in diabetic rats. AB - Upregulation of vascular NAD(P)H oxidase has been considered an important source for elevated levels of reactive oxygen species that contribute to several cardiovascular disease states, including the vascular complications of diabetes mellitus. Previous studies have shown that treatment with antioxidants corrects impaired nerve function and blood flow in diabetic rats. The aim was to assess the degree of involvement of NAD(P)H oxidase in experimental diabetic neuropathy. To this end, after 6 weeks of untreated streptozotocin-diabetes, rats were treated for 2 weeks with the NAD(P)H oxidase, apocynin. Two high doses (15 and 100 mg/kg) were used to ensure that maximal effects were registered. Diabetes caused a 20% reduction in sciatic nerve motor conduction velocity, and a 14% deficit for sensory saphenous nerve. Apocynin treatment corrected these defects by 32% and 48%, respectively: there were no significant differences between the effects of the 2 doses. Sciatic nerve nutritive endoneurial perfusion was measured by hydrogen clearance microelectrode polarography. Blood flow and vascular conductance were 47% and 40% reduced by diabetes, respectively. Both doses of apocynin had similar effects, correcting the blood flow deficit by 31% and conductance by 47%. Thus, the data show that NAD(P)H oxidase contributes to the neurovascular deficits in diabetic rats. While only accounting for part of the elevated reactive oxygen species production in diabetes, this mechanism could provide a novel therapeutic candidate for further investigation in diabetic neuropathy and vasculopathy. PMID- 12888121 TI - The role of inducible nitric oxide synthase inhibitor on the arteriolar hyporesponsiveness in hemorrhagic-shocked rats. AB - Hemorrhagic shock (HS) has been implicated in the induction of inducible nitric oxide synthase (iNOS) that leads to increase production of nitric oxide (NO). Recently, NO has been implicated to cause hyporesponsiveness of blood vessel in vitro towards vasoconstrictors in refractory (decompensated) HS. In our in vivo model, we examined the effects of aminoguanidine (AG), a known iNOS inhibitor, with angiotensin II (ANG II), a vasoconstrictor, following hemorrhagic shock decompensatory phase (HSDP) on percentage survival, vascular responsiveness, mean arterial blood pressure (MABP), heart rate and mean nitrate/nitrite levels in anaesthetized rats. HSDP (3 h) was achieved via constant pressure method (40-45 mmHg). MABP and heart rate was measured via the left carotid artery. Plasma collected from HSDP rats was used to measure nitrate/nitrite levels. Vascular hyporeactivity to ANG II was carried out using HSDP aortic strips, precontracted with KCl and noradrenaline. Sham-operated rats served as controls. HSDP rats decreased percentage survival, vascular contractility to ANG II and noradrenaline, MABP, heart rate while showing increased levels of nitrate/nitrite. Infusion of AG with ANG II, increased percentage survival and had reversed these cardiovascular effects of HSDP rats. This study indicates that excessive NO formation from iNOS activity induces vascular hyporeactivity and decompensation in HSDP. This might suggest that selective NOS inhibitor, AG, when coupled with ANG II, show reduction in NO's effect in HSDP. PMID- 12888122 TI - The mineralocorticoid receptor agonist, fludrocortisone, inhibits pituitary adrenal activity in humans after pre-treatment with metyrapone. AB - Whereas animal studies have shown a clear inhibitory effect of hippocampal mineralocorticoid receptors (MR) on hypothalamic-pituitary-adrenal (HPA) axis activity, investigations in humans revealed equivocal results. To further clarify the influence of MR in HPA activity we studied 10 healthy men during the circadian nadir of HPA activity (14:00 to 21:00) after pre-treatment with 3 g metyrapone to minimize the impact of basal endogenous cortisol secretion. On three separate occasions, in a placebo-controlled design, subjects received in a randomized order either 0.5 mg fludrocortisone p.o. or 0.2 mg aldosterone i.v. or placebo. Fludrocortisone exerted a significant inhibition of ACTH, cortisol and 11-desoxycortisol (p < 0.05), whereas no such effect was observed after aldosterone or placebo. These preliminary data suggest that MR are involved in the inhibition of the HPA axis during the circadian nadir of glucocorticoid concentrations in humans. PMID- 12888123 TI - Expression of mutant non-cleavable Fas ligand on retrovirus packaging cells causes apoptosis of immunocompetent cells and improves prodrug activation gene therapy in a malignant glioma model. AB - Recombinant retroviruses (RV) have been widely used as vectors for clinical gene therapy of malignant brain tumors. Because of the very limited stability of these vectors in vivo, RV producing cells (VPC) are routinely used for intratumoral RV release. The host immune system, however, recognizes the intratumorally grafted allogeneic or xenogeneic VPC, and mounts an immune response against them. Humoral and cellular immune responses eventually result in reduction of VPC numbers and in limited success of RV mediated gene therapy approaches. This study presents a non-pharmacological and spatially limited approach for protection of VPC grafted in the CNS against destruction by host immune responses. Murine fibroblast derived VPC expressing herpes-simplex-virus type I thymidine kinase (HSV-tk) were genetically modified to co-express a human Fas ligand (CD95L) deletion mutant (DeltaFasL) resistant to enzymatic cleavage and shedding. Direct interactions between Fas (CD95) on lymphocytes and DeltaFasL on VPC upon cell-cell contact rapidly caused apoptosis in lymphocytes. In addition, cultured malignant brain tumor cells (U87, LN18, LN229) transduced with DeltaFasL-RV were rendered apoptotic by Fas/DeltaFasL interaction. DeltaFasL-expressing VPC grafted in a 9L rat brain tumor model survived in significantly higher numbers compared with control VPC, and did not cause an increase in neutrophil infiltration of tumors. Gene therapy of tumor bearing animals grafted with the modified DeltaFasL-VPC and given the prodrug Ganciclovir resulted in significantly increased survival rates compared to treatment with control VPC and Ganciclovir. In conclusion, prolonged intratumoral presence of DeltaFasL-VPC seems to be a direct consequence of the expression of the membrane-bound mutant FasL, and may result in increased total RV output and improved tumor transduction with RV. PMID- 12888124 TI - Effect of i.c.v. infusion of the alpha-MSH agonist MTII on meal patterns in male rats following nicotine withdrawal. AB - The present study explored the role of endogenous alpha-MSH in the alteration of meal patterns induced by nicotine (NIC) withdrawal. Male Sprague Dawley rats bearing third ventricle cannulas were placed in computerized food intake monitors. On days 1-21, the rats were given 4 mg/kg/day of NIC or saline (SAL) in four equal i.p. doses during the dark period. NIC suppressed (P < 0.05) food intake only during the first week. The normalization of food intake occurred when the reduced meal size of the NIC injected rats was countered by an increase in meal number. Despite the normalization of 24-h food intake, body weight in NIC rats was decreased (P < 0.05) for 21 days. On day 22, the rats were divided into 4 groups (n's = 7-8 each) and injected into the third ventricle with various doses of the alpha-MSH agonist MTII or artificial cerebrospinal fluid (aCSF): SAL + aCSF, SAL + MTII, NIC + aCSF, NIC + MTII. Infusion of MTII (30 ng/rat) suppressed (P < 0.01) dark phase food intake in both groups, but the NIC + MTII group ate (P < 0.05) more than the SAL + MTII group. Meal number during the dark phase was suppressed by MTII, but the NIC + MTII group took significantly more meals that the SAL + MTII group. Infusion of MTII suppressed meal size in SAL and NIC treated rats, but this effect was attenuated in NIC treated rats. All meal parameters normalized by the day after i.c.v. infusion. These data indicate that NIC treatment differentially affects the neural controls of meal number and meal size and attenuates the suppression by MTII of meal number and meal size. PMID- 12888125 TI - Comparison of the prognostic value of RR-interval variability after acute myocardial infarction in patients with versus those without diabetes mellitus. AB - Diabetic patients have substantially greater long-term mortality after acute myocardial infarction (AMI) than nondiabetic patients. Traditionally, cardiac autonomic neuropathy is believed to contribute significantly to the increased mortality rate in patients with diabetes mellitus. In contrast, a recent study suggested that RR-interval variability (RRV) did not predict mortality after AMI in diabetic patients. We compared, in diabetic and nondiabetic patients, the relation between low RRV and long-term mortality in the Multicenter Post Infarction Program (MPIP), a longitudinal observational study of 715 survivors of AMI, including 117 diabetic patients. We studied the association between mortality and 6 frequency-domain measurements and 1 time-domain measurement of RRV. We tested 2 hypotheses: (1) RRV is lower in diabetic patients; and (2) low RRV is less predictive of mortality in diabetic patients. Reduced RRV was significantly more frequent in diabetic patients than nondiabetic patients for all measurements, except high-frequency (HF) power. In diabetic patients, the association between reduced RRV and long-term mortality was at least as strong as it was in nondiabetic patients for all measurements except HF power; this pattern was found for all-cause, cardiac, and arrhythmic death. In multivariable models, reduced RRV was significantly associated with all-cause mortality in diabetic patients even when adjusted for left ventricular ejection fraction, heart failure class, and ventricular arrhythmias. In our post-AMI sample, RRV provided valuable prognostic information among diabetic patients. Our findings suggest that cardiac autonomic neuropathy plays a role in the high mortality rate seen in diabetic patients after AMI. PMID- 12888126 TI - Comparison of the 80-lead body surface map to physician and to 12-lead electrocardiogram in detection of acute myocardial infarction. AB - Diagnosis of non-ST-elevation acute myocardial infarction (AMI) by a 12-lead electrocardiogram has poor sensitivity and specificity and, therefore, relies on biochemical markers of myocardial necrosis, which can only be reliably detected within 14 to 16 hours from symptom onset. The body surface map (BSM) improves AMI detection but is limited by its interpretation by inexperienced medical staff. To facilitate interpretation, an automated BSM algorithm was developed and is evaluated in this study. One hundred three patients with ischemic-type chest pain were recruited for this study from December 2001 to April 2002. A 12-lead electrocardiogram (Marquette Mac 5K) and BSM (PRIME-ECG) were recorded at initial presentation, and cardiac troponin I and/or creatine kinase-MB levels measured at 12 hours after symptom onset. The admitting physician's 12-lead electrocardiographic (ECG) interpretation, 12-lead ECG algorithm (Marquette 12 SL V233) diagnosis, and BSM algorithm diagnosis were documented for each patient. AMI, defined by elevation of troponin I to >1 microg/L and/or creatine kinase-MB to >25U/L, occurred in 53 patients. The admitting physician diagnosed 24 patients with AMI (sensitivity 45%, specificity 94%), the 12-lead ECG algorithm diagnosed 17 patients with AMI (sensitivity 32%, specificity 98%), and the BSM algorithm diagnosed 34 patients with AMI (sensitivity 64%, specificity 94%). The BSM algorithm improved the diagnostic sensitivity by 2.0 (p <0.001) and 1.4 (p = 0.002) compared with the 12-lead ECG algorithm or the admitting physician, respectively. There was no significant difference in specificity. Thus, the BSM algorithm improves detection of AMI compared with the 12-lead ECG algorithm or physician's 12-lead ECG interpretation. PMID- 12888127 TI - Comparison of patients with acute coronary syndrome with and without systemic hypertension. AB - The role of systemic hypertension in acute coronary syndrome (ACS) has not been well studied. We studied consecutive subjects admitted to the University of Michigan Health System (Ann Arbor, Michigan) with symptoms of ACS. Data were collected using a standardized form. This observational study is currently ongoing; we collected data from May 1999 to December 2000 for 979 subjects, 890 of whom also had 6-month follow-up data. Hypertensives represented 64.4% (n = 630) of the total population. In general, hypertensive patients were older than normotensives (66.3 vs 59.9 years, p <0.0001), more often women (38.7% vs 26.9%, p = 0.0002), and had more comorbidities, such as previous myocardial infarction (47.9% vs 33.8%, p <0.0001), congestive heart failure (25.7% vs 12.0%, p <0.0001), and diabetes (36.9% vs 17.8%, p <0.0001). At admission, hypertensives had higher systolic blood pressure. Hypertensives had fewer electrocardiographic abnormalities indicating ischemic changes (67.9% vs 76.3%, p = 0.01) and had fewer incident of acute myocardial infarction (AMI) (70.7% vs 76.1%, p = 0.07) than normotensives. There was consistency over different levels of admission systolic blood pressure. Hypertensives received more oral cardiovascular drugs, and had undergone more invasive procedures. The lower rate of AMI in hypertensives seemed to be related to the higher frequency of a history of percutaneous coronary intervention and coronary artery bypass grafting. However, at 6-month follow-up, age- and gender-adjusted odds ratios for adverse events were equivalent in hypertensives and normotensives, suggesting no continuing differential treatment benefit for hypertensives in the months after the initial ACS episode. PMID- 12888129 TI - Increased use of lipid-lowering therapy in patients receiving human immunodeficiency virus protease inhibitors. AB - Although human immunodeficiency virus (HIV) protease inhibitors (PIs) improve survival in patients with HIV infection, many patients receiving PIs develop hyperlipidemia, which may increase risk of future coronary events. The purpose of this study was to estimate the changing prevalence of lipid-lowering therapy (LLT) in patients with HIV and to evaluate its association with the use of HIV PIs. This was a cross-sectional study of adults with HIV infection who were registered in the Medicaid of California (MEDI-CAL) administrative claims database. Frequencies of HIV-related and dyslipidemia diagnoses were determined from International Classification of Diseases-9th Edition codes. Use of lipid lowering and antiretroviral medications was determined by National Drug Codes. Multivariate statistical techniques were used to evaluate trends in use of PIs and lipid-lowering medications from January 1996 to June 2002. The number of HIV infected patients in MEDI-CAL ranged from 15,764 in 1996 to 13,349 in 2000. The prevalence of LLT use among HIV-infected patients on PIs increased by sixfold (1.7% to 10.6%, p <0.05), and in 2000, exceeded use in the overall MEDI-CAL population (p = 0.09). The increasing rate of LLT in patients taking PIs was greater than in HIV-infected patients not on PIs and in MEDI-CAL (p = 0.002). In multivariate models, increasing age (odds ratio 2.30) and use of PIs (odds ratio 2.08) predicted use of LLT (p <0.001). Thus, in patients taking HIV PIs, use of LLT increased more than sixfold, at a faster rate than in the general population. It has not been proved that use of LLT in HIV-infected patients taking PIs improves survival. PMID- 12888130 TI - Relation between high-density lipoprotein cholesterol and peripheral vasomotor function. AB - Low levels of high-density lipoprotein (HDL) cholesterol are one of the most common lipid abnormalities in patients with coronary artery disease. Endothelial dysfunction is also highly prevalent in patients with coronary artery disease. We sought to determine whether HDL cholesterol levels are correlated with endothelium-dependent vasomotion in patients being evaluated for atherosclerosis. Peripheral vascular endothelial function was assessed by high-resolution brachial artery ultrasound. Flow-mediated dilation (FMD) during reactive hyperemia was defined as the percent change in arterial diameter following 5-minute arterial occlusion. All patients underwent stress testing with nuclear single-photon emission computed tomographic imaging to determine percent left ventricular ejection fraction and define the presence or absence of coronary artery disease. One hundred fifty-one subjects (87 men, 64 women) were enrolled (average age 58 +/- 11 years). Total cholesterol, HDL cholesterol, low-density lipoprotein cholesterol, and triglyceride levels were 188 +/- 48, 47 +/- 13, 108 +/- 37 and 154 +/- 88 mg/dl, respectively. The mean FMD for the entire group was 9.9 +/- 5.2%. Subjects with an HDL cholesterol of <40 mg/dl (n = 39) had lower FMD (7.4 +/- 3.6%) compared with those with an HDL cholesterol >/=40 mg/dl (11.0 +/- 5.5%, p <0.001). There was a significant correlation between FMD and HDL cholesterol level (linear regression, p <0.001), and in multivariate analysis, HDL cholesterol was an independent predictor of FMD. Peripheral endothelial function was abnormal in subjects with low HDL cholesterol and well-preserved in those with high HDL cholesterol. These data suggest that impaired endothelial function associated with low HDL cholesterol may be an additional, previously unrecognized mechanism contributing to the increased risk of atherosclerosis in these patients. PMID- 12888128 TI - Usefulness of prior hysterectomy as an independent predictor of Framingham risk score (The Women's Health Initiative). AB - The association of hysterectomy with increased coronary risk is controversial, and previous studies have reached differing conclusions as to whether the excess risk is confined to women who have also undergone bilateral oophorectomy. This analysis uses the Framingham algorithm to evaluate the hypothesis that hysterectomy with or without ovarian preservation is associated with increased coronary risk, using a cross-sectional analysis of baseline data from 1,501 participants of the Women's Health Initiative. Framingham risk scores, derived from the algorithm in the National Cholesterol Education Program Adult Treatment Panel III guidelines, which include age, smoking, systolic blood pressure, total and high-density lipoprotein cholesterol, were determined in a subgroup of Women's Health Initiative participants with measured plasma lipids and known ovariectomy status. Women with hysterectomy had fewer years of education than those without hysterectomy (30% with college degree vs 41%, p <0.0001) and higher body mass index (29 vs 28 kg/m(2), p <0.0001), consumed less alcohol, exercised less, and had a higher Framingham risk of myocardial infarction or coronary death (46% vs 41% with 10-year risk >/=4%, p = 0.04). In multivariate analysis, hysterectomy with bilateral oophorectomy was an independent predictor of Framingham risk (p = 0.04), whereas hysterectomy with ovarian preservation was not. PMID- 12888131 TI - ALLHAT--all hit or all miss? Key questions still remain. PMID- 12888132 TI - Left ventricular free wall rupture after reperfusion therapy for acute myocardial infarction. AB - We evaluated the clinical significance of angiographic indexes and pericardial involvement in predicting increased risk of free wall rupture after reperfusion therapy and found that Thrombolysis In Myocardial Infarction (TIMI) <3 flow grade after reperfusion therapy was a significant variable related to the free wall rupture. Moreover, pericardial rub was found to be a significant variable related to TIMI <3 grade flow after reperfusion, which indicates that detection of pericardial rub is one of the clinical signs that predicts inadequate anterograde flow of the infarct-related artery after reperfusion and hence, higher risk for free wall rupture. PMID- 12888133 TI - Comparison of clinical benefits of clopidogrel therapy in patients with acute coronary syndromes taking atorvastatin versus other statin therapies. AB - In clinical practice, we found no significant difference between atorvastatin therapy or other statin therapies in the clinical outcomes of patients with acute coronary syndromes receiving clopidogrel therapy. In patients receiving atorvastatin therapy, clopidogrel therapy was associated with a significant decrease in mortality and stroke during univariate analysis and a moderate trend of reduced mortality and stroke without statistical significance in the multivariate analysis. PMID- 12888134 TI - Effect of acute hyperglycemia on the ischemic preconditioning effect of prodromal angina pectoris in patients with a first anterior wall acute myocardial infarction. AB - Acute hyperglycemia abolishes the ischemic preconditioning effect of prodromal angina pectoris in patients with acute myocardial infarction. We investigated a potential explanation for the association between acute hyperglycemia and adverse outcomes after acute myocardial infarction. PMID- 12888135 TI - Effects of cardiac rehabilitation and exercise training on indexes of dispersion of ventricular repolarization in patients after acute myocardial infarction. AB - We studied 50 consecutive patients with relatively preserved systolic function (ejection fraction >40%, mean 53 +/- 11%) after acute myocardial infarction, and assessed indexes of dispersion of ventricular repolarization before and after a formal, phase II cardiac rehabilitation and exercise training program. After cardiac rehabilitation, statistically significant reductions occurred in QT dispersion, JT dispersion, and in the heart rate corrected indexes. These benefits add to the proven benefits of formal cardiac rehabilitation and exercise training programs and may reduce the subsequent risks of malignant ventricular arrhythmias and sudden cardiac death. PMID- 12888136 TI - Degrees of severe stenoses in sigma-shaped versus C-shaped right coronary arteries. AB - The right coronary artery (RCA) appears either C-shaped or sigma-shaped during standard angiography. The purpose of the present investigation was to assess whether C-shaped RCAs are associated with more atherosclerotic disease than sigma shaped RCAs. The study sample comprised 120 consecutive patients who underwent coronary catheterization and multivariate analysis was conducted using several systemic risk factors for atherosclerosis. The proportion of sigma-shaped RCAs found in a group whose angiograms showed little or no obstruction (70%) was significantly higher than that found in the group with significant obstruction (33%, p <0.001). In conclusion, a C-shaped RCA is associated with atherosclerosis. PMID- 12888137 TI - Anti-inflammatory effect of atorvastatin (80 mg) in unstable angina pectoris and non-Q-wave acute myocardial infarction. AB - In this randomized trial, C-reactive protein increased during the first 5 days of an acute coronary syndrome in patients treated with placebo, but this phenomenon was not observed in those randomized to atorvastatin 80 mg/day. This suggests that short-term statin therapy inhibits inflammation in patients with non-ST elevation acute coronary syndromes. PMID- 12888138 TI - Frequency of and outcome of acute coronary syndromes in patients with human immunodeficiency virus infection. AB - Fifty-one patients with human immunodeficiency virus infection and acute coronary syndromes were identified. Nearly all patients (98%) had traditional coronary risk factors. Revascularization procedures were performed safely with low in hospital mortality. PMID- 12888139 TI - Prevalence of left main coronary artery disease, of three- or four-vessel coronary artery disease, and of obstructive coronary artery disease in patients with and without peripheral arterial disease undergoing coronary angiography for suspected coronary artery disease. AB - Data from the present investigation showed that the prevalence of current cigarette smoking, current or ex-cigarette smoking, systemic hypertension, diabetes mellitus, and dyslipidemia was significantly higher in patients with peripheral arterial disease (PAD) than in patients without PAD. The present report also showed that compared with patients without PAD undergoing coronary angiography for suspected coronary artery disease (CAD), patients with PAD undergoing coronary angiography for suspected CAD had a higher prevalence of left main CAD (18% vs <1%), a higher prevalence of 3- or 4-vessel CAD (63% vs 11%), and a higher prevalence of obstructive CAD (98% vs 81%). PMID- 12888140 TI - Effects of statins on C-reactive protein and interleukin-6 (the Ludwigshafen Risk and Cardiovascular Health study). AB - This report provides preliminary evidence that statins may lower C-reactive protein levels by interfering with the generation and/or release of C-reactive protein in the liver rather than by modulating inflammatory processes in the vessel wall. PMID- 12888141 TI - Can the index of myocardial performance be used to detect acute cellular rejection after heart transplantation? AB - The index of myocardial performance (IMP), combining systolic and diastolic function, was measured in 50 orthotopic heart transplant recipients to determine if it could be used to detect acute rejection. It was calculated as the sum of the isovolumic contraction time (IVCT) and isovolumic relaxation time (IVRT) divided by the ejection time. Comparison of intrarecipient changes in Doppler intervals between rejection and nonrejection states demonstrated an increase in IVCT and decrease in IVRT during rejection with no significant change in the IMP. Rejection is likely to be best detected by assessing these parameters independently rather than by measuring the IMP. PMID- 12888142 TI - Usefulness of beta radiation for de novo and in-stent restenotic lesions in saphenous vein grafts. AB - Following successful angioplasty of 49 saphenous vein graft lesions, a novel, self-centering phosphorus-32 solid foil beta source encapsulated within a dual balloon membrane was used to deliver 20 Gy 1 mm into the vessel wall. Clinical and angiographic recurrence rates at 12 months were low, especially in de novo lesions. PMID- 12888143 TI - Long-term follow-up of patients after gamma intracoronary brachytherapy failure (from GAMMA-I, GAMMA-II, and SCRIPPS-III). AB - We report the long-term outcome of 225 patients who failed gamma-brachytherapy for in-stent restenosis. Total adverse events, target vessel revascularization, and myocardial infarction were higher after repeat percutaneous coronary intervention versus coronary artery bypass grafting. Therefore, coronary artery bypass grafting may be the preferable first-line therapy in these patients until other therapies (i.e., drug-eluting stents) are available. Shorter time from brachytherapy to radiation failure and late thrombosis after brachytherapy were independent predictors of adverse events. PMID- 12888144 TI - Relation between the degree of procedural anticoagulation and complications after coronary stent implantation. AB - The impact of the degree of anticoagulation in patients who underwent stent implantation without glycoprotein IIb/IIIa inhibitors was examined in 1,020 patients. High levels of procedural anticoagulation with heparin were found to increase hemorrhagic complications without improving clinical or angiographic outcomes. PMID- 12888145 TI - Relation of ischemia-modified albumin (IMA) levels following elective angioplasty for stable angina pectoris to duration of balloon-induced myocardial ischemia. AB - The results in this study confirm and expand previous reports that ischemia modified albumin (IMA) is an early marker of ischemia in the setting of percutaneous coronary intervention (PCI). We observed that IMA levels are related to the number of inflations, inflation pressure, and duration of inflations. It is therefore likely that IMA reflects the magnitude and duration of ischemia induced during PCI. PMID- 12888146 TI - Comparison of effects of a thrombin-based femoral artery closure device with those of a mechanical compression device on serum C-reactive protein and amyloid A after percutaneous coronary intervention. AB - The present study evaluates whether the closing procedure of the femoral artery after percutaneous coronary intervention influences the degree of inflammation related to the procedure as measured by C-reactive protein (CRP) and serum amyloid A (SAA). A thrombin-based device (Duett sealing device) was compared with a mechanical compression device (FemoStop). PMID- 12888147 TI - Effectiveness of sirolimus-eluting stent for treatment of left main coronary artery disease. AB - The present study reports on the clinical outcome of 31 consecutive patients with left main coronary artery disease treated with a sirolimus-eluting stent. The implantation of this stent was associated with abolition of post-discharge fatal events and percutaneous reintervention. PMID- 12888148 TI - Frequency of stent thrombosis after acute coronary syndromes (from the SYMPHONY and 2nd SYMPHONY trials). AB - We studied stent thrombosis in 4,607 patients with acute coronary syndromes who received a coronary stent as part of routine care during 2 trials of aspirin versus sibrafiban for secondary prevention. In these patients, stent thrombosis occurred more often than in previous patients who underwent elective percutaneous coronary intervention. These patients and their outcomes may be more representative of patients with typical acute coronary syndromes undergoing stenting in clinical practice. PMID- 12888149 TI - Relation of aggressiveness of lipid-lowering treatment to changes in calcified plaque burden by electron beam tomography. AB - The comparative effects of more versus less aggressive low-density lipoprotein (LDL) cholesterol lowering (to 80 mg/dl) on calcified coronary plaque progression by electron beam tomography were evaluated in 182 consecutive asymptomatic patients after 1.2 years of treatment with statins alone or in combination with niacin. Despite the greater improvement in lipids in the 80 mg/dl groups, there were no differences in calcified plaque progression (9.3%/year vs 9.1%/year). We conclude that, with respect to LDL cholesterol lowering, "lower is better" is not supported by changes in calcified plaque progression. PMID- 12888150 TI - Effect of atorvastatin and fenofibrate on autonomic tone in subjects with combined hyperlipidemia. AB - This randomized open-label trial investigated whether autonomic cardiovascular control is altered in middle-aged men with combined hyperlipidemia and whether such alterations are affected by short-term, lipid-lowering therapy with atorvastatin and/or fenofibrate. Compared with normolipidemic subjects, untreated subjects with combined hyperlipidemia had several abnormalities of autonomic tone, indicating increased sympathetic tone and decreased baroreflex sensitivity. The alterations in autonomic cardiovascular control were partially reversible by each of the lipid-lowering drugs. PMID- 12888151 TI - Reliability of QRS duration and morphology on surface electrocardiogram to identify ventricular dyssynchrony in patients with idiopathic dilated cardiomyopathy. AB - Using Fourier phase analysis of equilibrium radionuclide angiography in both ventricles, inter- and intraventricular delays were correlated with QRS morphology and duration in 103 patients with idiopathic dilated cardiomyopathy. We found that left bundle branch block on the left axis may reflect a high intra left ventricular dyssynchrony. A precise evaluation of the dyssynchrony appears useful because a high proportion of patients with incomplete bundle branch block, left anterior hemiblock, or "normal" QRS exhibit a marked intra-LV dyssynchrony and may respond to cardiac resynchronization therapy. PMID- 12888152 TI - Comparison of effects on systolic and diastolic left ventricular function of nebivolol versus atenolol in patients with uncomplicated essential hypertension. AB - This was a double-blind, randomized, prospective study comparing the effects of nebivolol and atenolol on systolic diastolic left ventricular function in patients with essential hypertension. A significant difference was seen in stroke volume between these 2 drugs. The blood pressure lowering effect of atenolol was strongly related to cardiac output and heart rate reduction. The blood pressure lowering effect of nebivolol was related to a reduction in peripheral resistance and an increase in stroke volume with preservation of cardiac output. This preservation of cardiac output, together with a reduced peripheral resistance, may be potentially important in treating heart failure. PMID- 12888153 TI - Usefulness of aortic valve calcium scores by electron beam computed tomography as a marker for aortic stenosis. AB - This study was undertaken to determine whether aortic valve calcium (AVC) scores measured by electron beam tomography can identify patients with echocardiographically defined aortic stenosis. Electron beam tomography is increasingly being used to detect coronary artery calcium. AVC can also be measured on electron beam tomographic (EBT) scans obtained to screen for coronary calcium. Whether EBT AVC scores correlate with the presence of aortic stenosis, as assessed by echocardiography, is unknown. Results of this study suggest that AVC scores should be calculated routinely for coronary calcium screening EBT scans, and that patients with Agatston AVC scores above a certain level (e.g., >150) should be referred for echocardiography. PMID- 12888154 TI - Late follow-up of nonoperative closure of secundum atrial septal defects using the King-Mills double-umbrella device. AB - There has been a marked increase in device closure of secundum atrial septal defects (ASDs) in the last decade. No clinical reports of late results have been forthcoming until now. The purpose of this report was to provide very long-term follow-up of nonoperative closure of secundum ADSs using the King-Mills Umbrella device. ASD closure using this technique did not appear to protect against the known secundum ASD complication of atrial arrhythmia. Twenty-seven-year follow-up of the umbrella ASD closure via the femoral vein shows effective occlusion, absence of device complications, and no reinterventions. PMID- 12888155 TI - Comparison of heart rate variability between surgical and interventional closure of atrial septal defect in children. AB - This study was designed to compare reduced heart rate variability (HRV) in children with atrial septal defect (ASD) after surgical ASD closure with interventional (Amplatzer device) closure. Reduced HRV was observed in all children with ASD before treatment. HRV was further impaired 1 month after surgical ASD closure but not after catheter device closure. Three months after treatment, HRV indexes tended to normalize in both groups. PMID- 12888156 TI - Comparison of left ventricular function by echocardiogram in patients with Wilms' tumor treated with anthracyclines versus those not so treated. AB - The aim of this report was to evaluate late left ventricular function in survivors of Wilms' tumor and to compare patients treated with anthracyclines with those treated without anthracycline and with normal subjects. Wilms' tumor survivors treated without anthracycline had no myocardial abnormalities. A large percentage of patients treated with anthracycline presented with increased end systolic wall stress. Results indicate that there is a high incidence of subclinical cardiovascular abnormalities in such patients. PMID- 12888159 TI - An evaluation of commonly employed central venous catheter kits and their potential risk for complications of excess guidewire introduction. AB - STUDY OBJECTIVE: To evaluate the components of commonly used central venous catheter kits with respect to the potential for guidewire-mediated complications during catheter placement.Prospective, nonrandomized, observational study. SETTING: Six academic hospitals across the United States. PATIENTS: None. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: 30 commercially manufactured catheter kits (15 tunneled, 15 nontunneled) were opened and evaluated. The catheter or sheath to be introduced was measured and a corresponding ideal guidewire length was calculated. The ideal length was then compared to the actual length, and differences were tabulated. Wire tip configuration and the presence and pattern of distance markings were recorded and, in conjunction with the catheter and wire length discrepancies, were used to grade the relative risk of introducing excess guidewire during catheter placement. Of 30 kits evaluated, 14 (46.7%) had guidewires more than 20 cm longer than necessary. The mean excess wire length was 15 cm (range: 8 to 55 cm) and did not differ significantly between tunneled and nontunneled catheter kits. Only 10 kits (33.3%) had distance markings of any type, and there was no standardization among them; none corresponded to previously published recommendations. There was potential risk of excess wire introduction in 18 catheter kits, of which seven were nontunneled devices designed for bedside placement. CONCLUSIONS: The design of commonly employed central venous access catheter kits is such that there is a mismatch between guidewire and catheter length and a general lack of guidewire markings. We believe that these designs may predispose to the introduction of excess guidewire and result in guidewire-mediated complications during catheter placement. This risk can likely be reduced by matching the guidewires to the devices placed over them and by standardizing guidewire distance markings. PMID- 12888158 TI - The relationship between ionized and total serum magnesium concentrations during abdominal surgery. AB - STUDY OBJECTIVE: To determine the necessity for ionized magnesium (iMg) assay by evaluating the effect of abdominal surgery without massive transfusion on total magnesium (Mg) and iMg concentrations. DESIGN: Prospective, observational study. SETTING: Operating rooms at a university teaching hospital. PATIENTS: 31 patients without electrolyte abnormalities who were scheduled for abdominal surgery. INTERVENTIONS: None. MEASUREMENTS: Serum electrolytes, iMg, ionized calcium, proteins before and after surgery, and urine Mg were recorded. MAIN RESULTS: Total Mg and iMg decreased: Mg 23%, iMg 15%; p < 0.0001. Total and iMg were highly correlated (r = 0.728, p < 0.0001). Urine Mg output remained within normal limits (mean 0.24 mmol/hr). CONCLUSIONS: During abdominal surgery without massive transfusion, both total and ionized hypomagnesemia occur. Changes in iMg and total serum Mg concentrations are closely correlated. Total serum Mg assay, while overestimating the prevalence of ionized hypomagnesemia, is sufficient to screen for hypomagnesemia. PMID- 12888160 TI - Rocuronium infusion requirements and plasma concentrations at constant levels of neuromuscular paralysis during three phases of liver transplantation. AB - STUDY OBJECTIVE: To develop a rapid online test of graft liver function during liver transplantation. DESIGN: Prospective, observational study. SETTING: University hospital transplant unit. PATIENTS: 17 adult patients with end-stage liver disease who underwent liver transplantation surgery. INTERVENTIONS: Rocuronium infusion dose requirements and plasma concentrations to maintain constant levels of neuromuscular paralysis during three phases of liver transplantation and their relationship with early postoperative liver function tests were studied. MEASUREMENTS: Infusion dose requirements of rocuronium, assay of rocuronium plasma concentrations using gas chromatography-mass spectrometry, and intensity of neuromuscular blockade were measured. MAIN RESULTS: A 24% decrease in rocuronium infusion requirements was observed during the an-hepatic phase. Rocuronium requirement during the neohepatic phase was increased only modestly or remained unchanged in 14 of the 16 patients who had normal graft function in the immediate postoperative period. Rocuronium plasma concentrations for maintaining constant levels of paralysis were significantly lower during the neohepatic phase than during the paleohepatic and anhepatic phases, indicating that there is likely to be a change in pharmacodynamics during this phase. Significant reduction in rocuronium infusion requirements during the neohepatic phase was observed in the only patient who had poor graft function in the early postoperative stage, suggesting that the reduced infusion requirement to maintain a constant neuromuscular paralysis may be related to the functional state of the graft liver after reperfusion. CONCLUSIONS: A significant reduction in rocuronium infusion requirement during the neohepatic phase may be suggestive of impaired organ function after reperfusion of the graft liver. Rocuronium may serve as a potential online indicator of graft liver function during liver transplantation by measurement of its infusion requirements during transplantation. PMID- 12888161 TI - Apnea and seizures following retrobulbar local anesthetic injection. AB - Surgery on the eye is performed using topical anesthesia, retrobulbar anesthesia, peribulbar anesthesia, and general anesthesia. Retrobulbar anesthesia is associated with a number of complications that include apnea (respiratory arrest), seizures, or both. Although these complications are transient and self limiting, they can be life-threatening if not recognized and treated early. We report two patients who developed apnea, one of whom had cardiorespiratory arrest; and two other patients who presented with seizures. We provided ventilation with 100% oxygen, treated the hypertension with nicardipine, and the tachycardia with esmolol. The patients did not have any residual complications. PMID- 12888162 TI - Epicardial echocardiography: diagnostic utility for evaluating aortic valve disease during coronary surgery. AB - Intraoperative epicardial echocardiography is commonly used to evaluate the ascending aorta for atheromatous disease before cannulation and cross clamping. In addition, it may serve as a cardiac imaging technique in patients where placement of a transesophageal echocardiography (TEE) probe is contraindicated, probe advancement is difficult, or a TEE probe is not available. We report a patient who was taken to the OR for coronary artery bypass grafting. Intraoperative TEE was planned to evaluate aortic valve function. However, attempts to place a TEE probe were abandoned due to high resistance on probe insertion. Epicardial echocardiography revealed previously undiagnosed aortic valve disease resulting in replacement of this valve. PMID- 12888164 TI - Another modified approach to the ganglion of Walther block (ganglion of impar). PMID- 12888163 TI - Pediatric death: guidelines for the grieving anesthesiologist. AB - This essay examines the effects of unexpected pediatric death on anesthesiology house staff, and offers a discussion of normal and abnormal patterns of grieving. The increased incidence of substance abuse and suicide among anesthesiologists is discussed, and the relationship of stress following patient death and appropriate coping skills is explored. A blueprint for managing stress is given based on a military combat stress model, and recommendations for residency training programs are made. PMID- 12888165 TI - Proficiency in interscalene anesthesia-how many blocks are necessary? AB - STUDY OBJECTIVE: To determine the number of interscalene blocks that are necessary for a resident in anesthesiology to complete so as to achieve autonomous success with that specific block. DESIGN: Survey questionnaire of all CA1 and CA2 residents in the Department of Anesthesiology, Mount Sinai School of Medicine. SETTING: University hospital. MEASUREMENTS AND MAIN RESULTS: Seventeen CA-1 and CA-2 residents reported their experiences in performing interscalene blocks over an 8-month period. Data were collected regarding the number of previous attempts, technique, attending supervisor, patient weight, level of autonomy in the performance of the current block, and adequacy of the block for surgery. Eighty-two blocks were performed (1 to >15 per resident), supervised by 7 attending physicians, and 96.3% of blocks were reported as adequate for surgery. With an experience of seven to nine previous blocks, only 50% of residents were able to perform an interscalene block autonomously, whereas with >15 blocks, 87.5% reported autonomous success. CONCLUSIONS: Experience with only 40 unspecified peripheral nerve blocks, as currently required by the Residency Review Committee as of January 2001, may not provide adequate opportunity for an individual to develop expertise in regional anesthesia. PMID- 12888166 TI - Recruitment of house staff into anesthesiology: a re-evaluation of factors responsible for house staff selecting anesthesiology as a career and individual training program. AB - STUDY OBJECTIVE: To reexamine, in a follow-up to our first study, those factors responsible for house staff (i.e., residents and clinical fellows) selecting anesthesiology as a career and a specific training program, as well as house staff satisfaction with various educational aspects of our training program, and their perceptions of the future for graduating anesthesiology trainees. DESIGN: Survey questionnaire of 77 house staff at the Mayo Clinic during the 2000 to 2001 academic year. SETTING: Academic medical center. MEASUREMENTS: A cross-sectional analysis was conducted using a questionnaire to survey 77 house staff enrolled in the anesthesiology training program at Mayo Clinic, Rochester, MN during the 2000 to 01 academic year. All responses were anonymous. Data were compared between time epochs using an f-exact test. A p-value 0.998) was obtained between the concentration and the detector signal. Recoveries were between 75 and 99% for the drugs and metabolites. The accuracy of the quality control samples, expressed as percent recovery, ranged from 91 to 118%; intra- and inter-assay-relative standard deviations were 0.9-10.2% and 0.9-9.7%, respectively. Additional external quality control is carried out since 3 years. This method is applicable to rapidly and effectively analyze serum or plasma samples for therapeutic drug monitoring of about 30 antidepressants and atypical antipsychotics. PMID- 12888197 TI - High-performance liquid chromatographic determination of quercetin in human plasma and urine utilizing solid-phase extraction and ultraviolet detection. AB - An HPLC method for determining quercetin in human plasma and urine is presented for application to the pharmacokinetic study of rutin. Isocratic reversed-phase HPLC was employed for the quantitative analysis by using kaempferol as an internal standard. Solid-phase extraction was performed on an Oasis HLB cartridge (>95% recovery). The HPLC assay was carried out using a Luna ODS-2 column (150 x 2.1 mm I.D., 5 microm particle size). The mobile phase was acetonitrile-10 mM ammonium acetate solution containing 0.3 mM EDTA-glacial acetic acid, 29:70:1 (v/v, pH 3.9) and 26:73:1 (v/v, pH 3.9) for the determination of plasma and urinary quercetin, respectively. The flow-rate was 0.3 ml/min and the detection wavelength was set at 370 nm. Calibration of the overall analytical procedure gave a linear signal (r>0.999) over a concentration range of 4-700 ng/ml of quercetin in plasma and 20-1000 ng/ml of quercetin in urine. The lower limit of quantification was approximately 7 ng/ml of quercetin in plasma and approximately 35 ng/ml in urine. The detection limit (defined at a signal-to-noise ratio of about 3) was approximately 0.35 ng/ml in plasma and urine. A preliminary experiment to investigate the plasma concentration and urinary excretion of quercetin after oral administration of 200 mg of rutin to a healthy volunteer demonstrated that the present method was suitable for determining quercetin in human plasma and urine. PMID- 12888198 TI - Determination of dextropropoxyphene and nordextropropoxyphene in urine by liquid chromatography-electrospray ionization mass spectrometry. AB - Dextropropoxyphene and nordextropropoxyphene were extracted from urine samples with mixed mode solid-phase extraction cartridges. After elution and evaporation to dryness, the eluate was dissolved in mobile phase and each sample was injected in a LC-ESI-MS system. Quantification was carried out in the selected ion monitoring mode. This article shows the possibility to analyse drugs of abuse substances in urine with a single quadrupole mass spectrometer if only a thorough work-up procedure and a sufficient chromatographic separation is accomplished. In order to enhance the fragmentation of the analytes, in-source fragmentation was carried out. One fragment and the pseudomolecular ion per analyte together with chromatographic retention times were sufficient to verify that the sought compound was found in the samples. In- and between day variation was lower than 10% and the recovery was well above 90%. The analytes were quantified in the range 100-10000 ng/ml urine. PMID- 12888199 TI - Purification of penicillin G acylase using immobilized metal affinity membranes. AB - The immobilized metal affinity membrane (IMAM) with modified regeneration cellulose was employed for purification of penicillin G acylase (PGA). For studying PGA adsorption capacity on the IMAM, factors such as chelator surface density, chelating metal, loading temperature, pH, NaCl concentration and elution solutions were investigated. The optimal loading conditions were found at 4 degrees C, 0.5 M NaCl, 32.04 micromol Cu(2+) per disk with 10 mM sodium phosphate buffer, pH 8.5, whereas elution conditions were: 1 M NH(4)Cl with 10 mM sodium phosphate buffer, pH 6.8. By applying these chromatographic conditions to the flow experiments in a cartridge, a 9.11-fold purification in specific activity with 90.25% recovery for PGA purification was obtained. Meanwhile, more than eight-times reusability of the membrane was achieved with the EDTA regeneration solutions. PMID- 12888200 TI - Quantification of 6-deoxy-6-demethyl-4-dedimethylaminotetracycline (COL-3) in human plasma using liquid chromatography coupled with electrospray ionization tandem mass spectrometry. AB - An accurate and reliable liquid chromatographic-tandem mass spectrometric (LC-MS MS) method has been developed and validated for the determination of 6-deoxy-6 demethyl-4-dedimethylaminotetracycline (COL-3) in human plasma. The assay used chrysin as an internal standard (I.S.). The analyte and the I.S. were extracted from acidified plasma by methyl-t-butyl ether. Separation was achieved on a YMCbasic column using acetonitrile-water-formic acid mobile phase. The MS-MS detection was by monitoring fragmentation 372.1-->326.2 (m/z) for COL-3 and 255.1 ->153.1 (m/z) for the I.S. on a Sciex API 365 using a Turbo Ionspray in positive ion mode. The retention times were approximately 1.7 min for COL-3 and 1.8 min for the I.S. The validated dynamic range was 0.03-10.0 microg/ml using 0.25-ml plasma with correlation coefficients of >or=0.9985. The precision and accuracy for the calibration standards (n=3) were RSD0.99). The lower limit of quantitation was 0.1 ng/ml. The assay was specific based on the multiple reaction monitoring transitions at m/z 1157-->215 and m/z 1101-->243 for thiocoraline and the internal standard, echinomycin, respectively. The mean intra- and inter-day assay accuracies remained below 5 and 12%, respectively, for all calibration standards and quality control (QC) samples. The intra- and inter-day assay precisions were less than 11.4 and 9.5% for all QC levels, respectively. The utility of the assay was demonstrated by a pharmacokinetic study of i.v. (bolus) thiocoraline on CD-1 mice. Thiocoraline was stable in mouse plasma in an ice-water bath for 6 h and for three freeze-thaw cycles. The reconstituted thiocoraline after extraction and drying sample process was stable in the autosampler for over 24 h. The assay was able to quantify thiocoraline in plasma up to 48 h following dose. Pharmacokinetic analysis showed that thiocoraline has distinct pharmacokinetic profiling when dosed in different formulation solutions. The assay is currently used to measure thiocoraline plasma concentrations in support of a project to develop a suitable formulation with a desirable pharmacokinetic profile. PMID- 12888202 TI - Determination of the enantiomers of ketamine and norketamine in human plasma by enantioselective liquid chromatography-mass spectrometry. AB - A sensitive enantioselective liquid chromatographic assay with mass spectrometric detection has been developed and validated for the simultaneous determination of plasma concentrations of (R)- and (S)-ketamine, and (R)- and (S)-norketamine. The compounds were extracted from human plasma using solid-phase extraction and then directly injected into the LC-MS system for detection and quantification. Enantioselective separations were achieved on a liquid chromatographic chiral stationary phase based upon immobilized alpha(1)-acid glycoprotein (the Chiral AGP column). The separations were achieved using a mobile phase composed of 2 propanol-ammonium acetate buffer (10 mM, pH 7.6) (6:94, v/v), a flow-rate of 0.5 ml/min and a temperature of 25 degrees C. Under these conditions, the analysis time was 20 min. Detection of the ketamine, norketamine and bromoketamine (internal standard) enantiomers was achieved using selected ion monitoring at m/z 238.1, 224.1 and 284.0, respectively. Extracted calibration curves were linear from 1 to 125 ng/ml per enantiomer for each analyte with correlation coefficients better than 0.9993 and intra- and inter-day RSDs of less than 8.0%. The method was applied to samples from a clinical study of ketamine in pain management. PMID- 12888203 TI - Purification and characterization of a small (7.3 kDa) putative lipid transfer protein from maize seeds. AB - The present study reports, for the first time in literature, the purification and biochemical characterization of a small basic protein from maize seeds similar to plant lipid transfer proteins-2, named mLTP2. The mLTP2 consists of 70 amino acid residues and has an M(r) of 7303.83, determined by electrospray ionization mass spectrometry. The primary structure of mLTP2 was determined by automated Edman degradation of the intact protein and peptides obtained from digestions with trypsin and by C-terminal sequencing using carboxypeptidase Y. The mLTP2 exhibits high sequence similarity (51-44% identical positions) with other plant LTP2s previously described. PMID- 12888204 TI - Quantification of melatonin in human saliva by liquid chromatography-tandem mass spectrometry using stable isotope dilution. AB - A method for the determination of melatonin in human saliva has been developed using high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS MS). Saliva was collected in plastic tubes. 7-D-Melatonin was added as internal standard and the samples were cleaned and concentrated by solid-phase extraction. The limit of detection was 1.05 pg x ml(-1) and the limit of quantification was 3.0 pg x ml(-1). The accuracy of the method was +/-14% at 5.60 pg x ml(-1) and +/ 9% at 19.6 pg x ml(-1). The precision was +/-13% at 6.18 pg x ml(-1) and +/-11% at 31.2 pg x ml(-1), respectively. Our HPLC-MS-MS method shows a high sensitivity and specificity for melatonin and more reliable results compared with a radioimmunoassay. The chromatographic method has been used to determine the circadian rhythm of melatonin among three nurses working the night shift and a patient suffering from an inability to fall asleep at night. PMID- 12888205 TI - Development of a liquid chromatography-mass spectrometry method for the high accuracy determination of creatinine in serum. AB - An LC-MS method for the high-accuracy determination of creatinine in serum has been developed and used to provide results for an international measurement evaluation programme (IMEP) and the Comite Consultatif pour la Quantite de Matiere (CCQM) international inter-laboratory studies. An assessment of different sample preparation methods, including ion-exchange chromatography, solid-phase extraction, plasma ultrafiltration and ethanol protein precipitation, revealed that no bias or reduced precision was associated with the quicker less extensive clean-up methods, when using liquid chromatography-isotope dilution mass spectrometry (LC-IDMS) for quantitation. A number of different calibration regimes were also investigated. External calibration was shown to provide adequate calibration for most routine analysis with a relative associated expanded uncertainty (k=2) of 6% at the 95% confidence level. The use of a non isotopically labelled internal standard was shown to improve the relative expanded uncertainty (k=2) to 4%. However, the difference in retention time between the internal standard and the creatinine was such that a matrix interferent produced an observed bias of over 16%. The use of an isotopically labelled internal standard was shown to reduce any bias to less than 0.2% with an expanded uncertainty (k=2) of less than 0.3%. The developed method was then used, in a blind trial organised jointly by IMEP and CCQM, to determine the amount of creatinine in human serum. The method performed well against the established reference method of ion-exchange chromatography followed by derivatisation gas chromatography (GC)-IDMS. The observed difference between the values determined by LC-IDMS and the key comparison reference value (average of all the submitted results) was less than 0.3%. The biggest advantage of the described method is in the speed of analysis. With a chromatographic run time of less than 10 min and sample preparation consisting of a simple protein precipitation, without the need for a derivatisation stage, the analysis is vastly simpler then the conventional GC-IDMS reference method. PMID- 12888206 TI - New high-resolution mass spectrometric approach for the measurement of polychlorinated biphenyls and organochlorine pesticides in human serum. AB - To increase our analytical throughput for measuring polychlorinated biphenyls (PCBs) and organochlorine (OC) pesticides without sacrificing data quality, we have developed and validated a combined PCB/OC pesticide gas chromatography-high resolution mass spectrometry (GC-HRMS) analysis. In a single GC-HRMS analysis, both selected PCBs and OC pesticides are detected and quantified. Previously, this has been difficult, if not impossible, because of the major difference in masses of the most abundant electron-impact ions. However, we have identified slightly less abundant ions to monitor that allow us to successfully combine these analytes into a single analysis without sacrificing any analytical sensitivity or instrument reliability. Consequently, we have been able to double our analytical throughput by modification of mass spectrometric parameters alone. Our new methodology has been validated against our current GC-HRMS method, which entails using two separate injections, one for PCB analysis and one for OC pesticide analysis. The two methods differ by less than 4% overall, with no systematic bias. We used this method to analyze approximately 350 serum samples over a period of several months. We found that our new method was as reliable in automated, overnight runs as our current method. PMID- 12888207 TI - Enzyme assay for protein kinase using micellar electrokinetic chromatography with laser-induced fluorescence detection. AB - Micellar electrokinetic chromatography (MEKC) with laser-induced fluorescence (LIF) detection has been developed for a protein kinase assay. This protein kinase assay could readily determine the phosphorylation activity of substrate peptide kemptide using cAMP-dependent protein kinase (PKA) as a model enzyme. Kemptide and phosphorylated kemptide could be reacted with 7-fluoro-4-nitrobenzo 2-oxa-1,3-diazole (NBD-F) as a fluorescence derivatization reagent for LIF detection by directly adding NBD-F into the PKA enzymatic reaction mixture. These derivatives of substrate and product were separated and detected within the analysis time of 5 min by micellar electrokinetic mode using a mixture of sodium dodecylsulfate and methanol as a running buffer. Good linearity of the peak response of the phosphorylated kemptide was obtained over the range of 1-20 mU/tube of PKA in the assay. The relative standard deviation of the peak areas of the phosphorylated kemptide using 2, 5 and 10 mU/tube of PKA were calculated to <10.4%, indicating that the assay was reproducible. Also, IC(50) values of six PKA inhibitors, the K(i) value and the inhibition pattern of one inhibitor, which were calculated to estimate by the variation of the peak area of the phosphorylated kemptide using 5 mU/tube of PKA, were consistent with the published data. The sensitivity of the assay was higher than that of enzyme linked immunosorbent assay (ELISA) for PKA phosphorylation activity, as IC(50) values, K(i) value, and the inhibition mechanism of inhibitors could be estimated using one-tenth amounts of PKA, compared with that of ELISA. The MEKC-LIF is expected to be very useful for protein kinase assay and its application to the estimation of inhibitors because this method does not entail experimentally troublesome procedures such as the preparation of antibody or fluorescence labeled substrate. PMID- 12888208 TI - Validation of a simple liquid chromatography assay for creatine suitable for pharmacokinetic applications, determination of plasma protein binding and verification of percent labeled claim of various creatine products. AB - Creatine has been quantified in various tissues by a range of methodologies. This paper reports on the development and validation of a simplified HPLC assay to determine plasma creatine, plasma protein binding of creatine, creatine in microdialysate and creatine in over-the-counter products. An isocratic, reversed phase (C(18)) HPLC assay, using potassium phosphate monobasic (pH 4) as a mobile phase, was validated in human plasma and microdialysis perfusion fluid (normal saline). The lower limit of quantification for the assay was 1 mg l(-1) in saline and 5 mg l(-1) in plasma. The RSD was below 6% and accuracy was below 12% in both matrices. Protein binding in human plasma was found to be negligible (<10%). Over the-counter creatine monohydrate products tested contained 100% creatine monohydrate. This assay was found to be suitable for pharmacokinetic studies and the assessment of plasma creatine and skeletal muscle microdialysate. PMID- 12888209 TI - Simultaneous determination of JTT-501 and its main metabolite in human plasma by liquid chromatography-ionspray mass spectrometry. AB - An LC-MS-MS analytical method was developed for the determination of a new antidiabetic agent, JTT-501 and its main metabolite (JTP-20604) in human plasma. The compounds were isolated from plasma by protein precipitation before analysis by HPLC with atmospheric pressure positive ionisation MS-MS detection. An isotopically labelled analog of JTT-501 was used as the internal standard. Linearity was demonstrated over the calibration range of about 5-10000 ng/ml for both compounds. The assay was validated with respect to accuracy, precision and analyte stability. This method was used for the determination of plasma concentrations for the two compounds in a clinical tolerability study. A cross validation exercise between two different mass spectrometers, used for the determination of clinical samples, is also reported. PMID- 12888210 TI - Fast and sensitive determination of urinary 1-hydroxypyrene by packed capillary column switching liquid chromatography coupled to micro-electrospray time-of flight mass spectrometry. AB - The present work reports capillary liquid chromatographic column switching methodology tailored for fast, sensitive and selective determination of 1 hydroxypyrene (1-OHP) in human urine using micro-electrospray ionization time-of flight mass spectrometric detection. Samples (100 microl) of deconjugated, water diluted and filtered urine samples were loaded onto a 150 microm I.D.x 30 mm 10 microm Kromasil C(18) pre-column, providing on-line sample clean-up and analyte enrichment, prior to back flushed elution onto a 150 microm I.D.x 100 mm 3.5 microm Kromasil C(18) analytical column. Loading flow rates up to 100 microl/min in addition to the use of isocratic elution by a mobile phase composition of acetonitrile/water (70/30, v/v) containing 5 mM ammonium acetate provided elution of 1-OHP within 5.5 min and a total analysis time of less than 15 min with manual operation. Ionization was performed in the negative mode and 1-OHP was observed as [M-H](-) at m/z 217.08. The method was validated over the concentration range 0.2-40 ng/ml 1-OHP in pre-treated urine, yielding a coefficient of correlation of 0.997. The within-assay (n=6) and between-assay (n=6) precisions were in the range 6.4-7.3 and 7.0-8.1%, respectively, and the recoveries were in the range 96.2-97.5 within the investigated concentration range. The method mass limit of detection was 2 pg, corresponding to a 1-OHP concentration limit of detection of 20 pg/ml (0.09 nmol/l) diluted urine or 0.3 ng/ml (1.35 nmol/l) urine. PMID- 12888211 TI - On-line coupling of solid-phase extraction with mass spectrometry for the analysis of biological samples. III. Determination of prednisolone in serum. AB - Solid-phase extraction (SPE) was directly coupled to mass spectrometry (MS) to assess the feasibility of the system for the rapid determination of prednisolone in serum. A C(18) stationary phase allowed washing of the cartridge with 25% methanol. Elution was performed by switching the methanol percentage from 25% in the washing step to 50% during elution. The high flow-rates during the extraction (5.0 ml/min) combined with ion-trap MS detection resulted in a total analysis time of 4 min. Some tailing of the prednisolone peak was observed. However, the tailing was found acceptable, since by this elution procedure most matrix compounds were prevented from eluting from the cartridge. Some matrix interference was still observed with a triple-quadrupole MS, even in the multiple reaction monitoring mode. This resulted in a detection limit (LOD) of about 10 ng/ml. The matrix interference and the LOD were similar for atmospheric pressure chemical ionisation and atmospheric pressure photo ionisation. Applying an ion trap MS in the MS-MS mode resulted in cleaner chromatograms. Due to extensive fragmentation of prednisolone, the LOD was not lower than about 5 ng/ml prednisolone in serum, and a limit of quantitation of about 10 ng/ml (relative standard deviation <15%) was observed. PMID- 12888212 TI - Application of high-performance liquid chromatography-mass spectrometry to detection of diuretics in human urine. AB - A rapid, sensitive and reliable high-performance liquid chromatographic-mass spectrometric method for the detection of 25 diuretics in human urine has been developed. Atmosphere pressure chemical ionization (APCI) and electrospray ionization (ESI) modes were evaluated. A 2-ml volume of urine was extracted under basic conditions and separated on an Agilent Zorbax SB-C(18) column (150 x 2.1 mm, 5 microm). The mobile phase consisted of formic ammonium-formic acid buffer (pH 3.5) and acetonitrile. The effects of capillary temperature, sheath gas pressure and compositions of mobile phase on the sensitivity were studied. The recoveries of most of the diuretics were 75-95%. In the full scan mode, the limits of detection of the 25 diuretics were 0.25-25 ng/ml for APCI and 0.6-250 ng/ml for ESI. Under the optimal conditions, 14 diuretics from authentic urine samples were detected successfully by LC-APCI-MS. To obtain more fragmentation information on the chemical structure for positive confirmation, tandem mass analysis was also investigated. PMID- 12888213 TI - The morphological and biochemical response of avian embryonic sympathoadrenal cells to nerve growth factor is developmentally regulated. AB - Cellular differentiation is a stepwise process where environmental factors are essential key components to direct cells to their final phenotype. The sympathoadrenal (SA) system is one of the principal models used to study the role of environmental factors in the development of the peripheral nervous system. Two major cell types originate from the SA progenitor: the principal neurons of the sympathetic ganglia and the chromaffin cells of the adrenal medulla. These cells are directed to their final phenotype by a series of environmental factors, of which nerve growth factor (NGF) and glucocorticoids, are the best studied. Previously, we have shown that 11-day embryonic chick sympathetic cell cultures increased their neuropeptide Y (NPY) protein and mRNA levels in the presence of NGF. In contrast, NGF had no such effect in chromaffin cell cultures from the same developmental stage. These results were unexpected since both cells types respond morphologically to NGF. To determine if these cells can gain or lose their capacity to respond to NGF, morphological and biochemical studies were done at earlier stages using immunocytochemical, radioimmunoassay and polymerase chain reaction (PCR) techniques. Interestingly we found that in E-7 chromaffin cells there is a biochemical and morphological response to NGF, while E-7 sympathetic cells lack this response. Our observations show a developmental point of regulation of morphological and biochemical properties by NGF and reveal an age dependent capacity of SA cells to acquire or lose competence to an environmental factor. PMID- 12888214 TI - Identity and neuroanatomical localization of messenger RNAs that change expression in the neural tube of mouse embryos within 1 h after ethanol exposure. AB - Changes in the expression of various species of messenger RNA (mRNA) are likely to be one of the initial responses of an embryo to in utero ethanol exposure. However, the identity and range of the initial species of mRNAs that change expression in an embryo after acute prenatal ethanol exposure are unknown. mRNA was isolated from embryos treated on embryonic day 9.5 and examined 1 h after treatment. Ethanol-exposed embryos were compared to similarly treated maltose/dextrin and no injection control embryos. Subtractive hybridization was used to find the species of mRNAs. The identity of the differentially expressed mRNAs was ascertained using standard sequencing techniques while in situ hybridization defined the anatomical localization of each message. Four messages from ethanol-treated embryos were differentially expressed. One message has no homology to any known gene in GenBank while three messages are homologous to known genes: the transmembrane molecule mh19, the heterologous ribonuclear protein hnRNP A3, and the mitochondrial enzyme AK2. One message (the novel gene) was turned on, while mh19, AK2 and hnRNP A3 were up-regulated in ethanol-exposed embryos. All messages were expressed in the developing neural tube of the ethanol exposed embryos, although there were differences in regional localization and extent of expression. Thus, the initial response of the embryo is rapid with changes in expression of multiple genes observed within 1 h after ethanol exposure. Further, these differentially expressed mRNAs are candidates for modulating the type and extent of ethanol-induced damage in the developing CNS. PMID- 12888215 TI - Neuroanatomical development in the absence of PKC phosphorylation of the myristoylated alanine-rich C-kinase substrate (MARCKS) protein. AB - The myristoylated alanine-rich C-kinase substrate protein (MARCKS) is a widely expressed target of protein kinase C (PKC) phosphorylation. Disruption of Marcks in mice leads to a number of developmental defects within the central nervous system that are completely prevented by expression of an epitope-tagged wild-type human MARCKS transgene. In the present study, we investigated whether PKC phosphorylation of MARCKS is necessary for normal central nervous system development and postnatal survival. Expression at approximately twice normal levels of a mutant MARCKS protein in which the four PKC phosphorylatable serines were replaced by asparagines did not allow postnatal survival of Marcks(-/-) pups. Nonetheless, the rescued animals exhibited none of the characteristic anatomical defects seen in the brains and retinas of knockout mice, suggesting that PKC phosphorylation of MARCKS is not required for normal central nervous system development. Expression studies showed that transgene expression was limited to the central nervous system, which has implications for the lack of postnatal survival as well as for the pathogenesis of the neuronal ectopia characteristic of MARCKS deficiency. A novel aspect of the MARCKS-deficient phenotype was also noted, absence of the pontine nuclei; this was also largely reversed in Marcks(-/-) animals expressing the mutant transgene. These data raise the possibility of a role for MARCKS in the netrin-regulated process of pontine nuclei formation. PMID- 12888216 TI - Moderate alcohol exposure compromises neural tube midline development in prenatal brain. AB - We previously reported that fetal alcohol treatment compromised the development of the midline raphe and the serotonin neurons contained in it. In this study, we report that the timely development of midline neural tissue during neural tube formation is sensitive to alcohol exposure. Pregnant dams were treated from embryonic day 7 (E7, prior to neurulation) or E8.5 (at neurulation) with the following diets: (a) alcohol (ALC), given as either a 20% or 25% ethanol-derived calorie (EDC) liquid diet, or (b) isocaloric liquid diet pair-fed (PF), or (c) standard rat chow (Chow). Fetal brains from each group were examined on E13, E15, or E18. Neural tube development was compromised as a result of alcohol exposure in the following ways: (1) approximately 60% of embryos at E13 and 20% at E15 showed perforation of the floor plate in the diencephalic vesicle, (2) although completely closed at E13, 70-80% of embryos failed to complete the formation of neural tissue at the roof as the alcohol exposure continued to E15, and (3) 60 80% of embryos show delayed 'occlusion' of the ventral canal by newly formed nestin-positive neuroepithelial cells and S100beta-positive glia in the brainstem of E15. The compromised (incomplete) neural tube midline (cNTM) occurred near the ventricles at E13 and E15, but was later completed at E18. In all cases, the cNTM was accompanied by an enlarged ventricle, and dose-dependent brain weight reduction. The midline of the neural tube at the roof and floor plates is known to mediate timely trophic induction for neural differentiation. Prenatal midline deficits also have the potential to affect the development of midline neurons such as raphe, septal nuclei, and the timely crossing of commissural fibers. The results of the liquid diet alcohol exposure paradigm suggest it is more a model for Alcohol-Related Neurodevelopmental Disorder (ARND) featuring neuropsychiatric disorders than for full-blown fetal alcohol syndrome (FAS) with noticeable facial dysmorphogenesis and gross brain retardation. PMID- 12888217 TI - Glycine facilitates transmitter release at developing synapses: a patch clamp study from Purkinje neurons of the newborn rat. AB - Synaptic currents in immature Purkinje cells from rats on postnatal days 0-14 (P0 P14) were studied using whole-cell patch-electrodes applied to cerebellar slices (200 micro m in thickness). Purkinje cells (held at -40 mV) showed excitatory postsynaptic currents (EPSCs) and inhibitory postsynaptic currents (IPSCs) spontaneously. From P2 to P12 the frequencies of miniature EPSCs and miniature IPSCs in the Purkinje cells increased by 10-fold or more, suggesting progressive formation of functional synapses during this period. Application of glycine (100 micro M) to an immature Purkinje cell at P3-10 immediately increased the frequencies of both EPSCs and IPSCs. The effects of glycine showed maximum at P5 6 for EPSCs and at P9-10 for IPSCs and decreased thereafter. Facilitatory effects of glycine were suppressed by strychnine (1 micro M), a specific blocker of the ionotropic glycine receptor, while the effects were also induced by other glycinergic agonists, including alpha-L-alanine (1 mM), L-serine (1 mM) and taurine (500 micro M). The site of glycinergic effects was studied by removing the action potential generation in cerebellar slices. Following the addition of tetrodotoxin (TTX, 1 micro M), the glycine-induced facilitation of EPSC almost disappeared, while that of IPSC remained (i.e. miniature IPSCs) and reached more than half of the value without TTX. These findings suggest that the ionotropic glycinergic receptors are expressed transiently but profoundly in the developing cerebellum, and that the distributions of these receptors causing excitation are different at excitatory and inhibitory presynaptic neurons. The glycine receptors may play distinct roles in the maturation and organization of cerebellar neural circuits. PMID- 12888218 TI - Involvement of intracellular calcium and protein phosphatases in long-term depression of A-fiber-mediated primary afferent neurotransmission. AB - Long-term depression (LTD) of monosynaptic and polysynaptic excitatory postsynaptic potentials (EPSPs) in substantia gelatinosa (SG) neurons can be induced by brief high-frequency electrical stimulation (HFS, 300 pulses at 100 Hz) of primary afferent fibers in dorsal roots. Here we examined the possible cellular mechanism underlying spinal LTD. Conventional intracellular recordings were made from SG neurons in a transverse slice-dorsal root preparation of the young rat lumbar spinal cord. LTD of both monosynaptic and polysynaptic EPSPs was induced in 16 of 24 SG neurons by HFS of dorsal root in either the presence or absence of the GABA(A) receptor antagonist bicuculline and the glycine receptor antagonist strychnine. Loading the postsynaptic cell with BAPTA, an intracellular Ca(2+) chelator, almost completely blocked the induction of LTD. Induction of LTD was abolished by bath application of calyculin A (100 nM), a potent inhibitor of protein phosphatases 1 and 2A. These results indicate that: (i) a rise in postsynaptic Ca(2+) is necessary for LTD induction, (ii) synaptic activation of protein phosphatases 1 and 2A plays an important role in the induction of LTD of primary afferent A-fiber neurotransmission in the young rat spinal cord, and (iii) the effect of LTD may be physiologically relevant for transmission and integration of sensory information, including nociception. PMID- 12888219 TI - Activity-dependent neuroprotective protein: a novel gene essential for brain formation. AB - We have recently cloned the novel homeobox-containing activity-dependent neuroprotective protein (ADNP). In the current study, mouse ADNP was shown to be expressed at the time of neural tube closure, detected at E7.5 and increased on E9.5. Expression was augmented in the brain (E12.5), sustained throughout embryogenesis and regulated by VIP. To assess the function of ADNP, knockout mice were established. Detailed analysis revealed cranial neural tube closure failure and death on E8.5-9.0 of the ADNP-knockout embryos. The expression of Oct4, a gene associated with germ-line maintenance was markedly augmented in the knockout embryos. In contrast, the expression of Pax6, a gene crucial for cerebral cortex formation, was abolished in the brain primordial tissue of the knockout embryos. Thus, Pax6 and Oct4 constitute a part of the mechanism of action of ADNP on brain formation, inhibiting germ-line division while activating morphogenesis. In conclusion, ADNP is identified here as a new key gene essential for organogenesis in the developing embryo and may be implicated as a clinical target associated with proper neurodevelopment. PMID- 12888220 TI - Late differentiation contributes to the apparent increase in sensory neuron number in juvenile rat. AB - Using both profile counts and unbiased stereological methods, estimates of neuron number in the lumbar dorsal root ganglia of the rat have been shown to increases approximately 35% during postnatal life [J. Comp. Neurol. 386 (1997) 8-15; J. Comp. Neurol. 449 (2002) 158-165]. The mechanism underlying this addition of neurons was investigated. No evidence of incorporation of (BrdU), a mitotic marker, was found. Similarly, counts of myelinated and unmyelinated axons in the sural nerve were the same in neonates and adults. These results are not consistent with the possibility that neurogenesis accounts for neuron addition. A population of neurons that stains with TuJ1, an antibody against neuronal class III beta tubulin, but not with an antibody against the phosphorylated and non phosphorylated forms of heavy chain neurofilament protein (NF-H) was found in neonates, but not adults. These less-differentiated (type-L) neurons are not detected by either profile counts or unbiased stereology and do not transport HRP retrogradely. Maturation of this pool of incompletely differentiated neurons appears to be one mechanism whereby neuron number is augmented during postnatal life. PMID- 12888221 TI - The permeation and activation properties of brain sodium channels change during development. AB - BTX-modified sodium channels from 15-day embryonic (E15) rat forebrains were studied in planar lipid bilayers. Compared to postnatal sodium channels, E15 channels had a lower maximal single channel conductance, whereas their permeation pathway sensed a comparable surface charge density and had a similar apparent binding affinity for sodium ions. The steady-state activation curve of E15 channels was significantly more hyperpolarized and had a shallower slope than postnatal channels. The apparent BTX binding affinity was significantly lower for E15 channels than for postnatal channels. Finally, E15 channel alpha-subunits displayed a lower apparent molecular weight, and a lower sialylation level than postnatal sodium channel alpha-subunits. Together with previous studies, our data suggested that the observed functional differences between E15 and postnatal voltage-dependent sodium channels cannot be explained solely by the observed differences in channel sialylation, and hence they also appeared to reflect the presence of other channel structural differences. PMID- 12888222 TI - Prenatal development of peptidergic primary afferent projections to mouse lumbosacral autonomic preganglionic cell columns. AB - To examine the prenatal development of spinal visceral reflexes, primary sensory nerve fibers immunoreactive for calcitonin gene-related peptide (CGRP) were examined in the spinal cord, particularly in the autonomic preganglionic nuclei of mouse embryos. On embryonic day 16 (E16), CGRP-immunoreactive fibers were first observed in the sacral intermediolateral nucleus (IML) of the parasympathetic division as well as in the lumbar central autonomic nucleus (CA) of the sympathetic division, where they appeared in proximity to preganglionic neuronal perikarya immunoreactive for choline acetyltransferase or nitric oxide synthase. Most of the CGRP-immunoreactive varicosities were negative for substance P. Substance P-immunoreactive varicosities were scattered in these nuclei, but no appositions were seen on the preganglionic neuronal perikarya. On E18, CGRP-immunoreactive fibers were more abundant in the sacral IML and the lumbar CA. Co-expression of substance P and CGRP was frequently observed in the varicosities very close to the preganglionic neuronal perikarya on E18. CGRP immunoreactive fibers were also observed in the lumbar IML on E18, although significantly fewer were found in this nucleus compared with the sacral IML. In contrast to the upper lumbar level, no fibers immunoreactive for CGRP were observed in the IML at the thoracic level. These results suggest that peptidergic primary sensory fibers grow to project to the selective targets of autonomic preganglionic neurons during the embryonic period. The potential direct connections between the peptidergic primary sensory fibers and preganglionic neurons innervating the pelvic viscera might provide a circuit for spinal visceral reflexes active in embryos. PMID- 12888223 TI - Accident profile of older people in Antalya City Center, Turkey. AB - Accidents are major health problems leading to deaths and injuries among older people. The present study was performed to investigate the characteristics of the accidents experienced within the last 1 year in people aged 60 years and older living in Antalya City Center. The study was planned as a cross-sectional research. A total of 840 individuals selected from the study population with cluster-sampling method were used in questionnaires. A number of 163 (19.4%) individuals had at least one accident in the last year. A total of 178 accidents were reported within the last 1-year; 124 (69.7%) falls, 22 (12.4%) traffic accidents and 12 (6.7%) dropping of objects to head. The accidents occurred mostly at home (40.4%), at avenue-street etc. (31.5%), and in garden (8.4%). The result of logistic regression analysis revealed that accident frequency was positively related with female gender (odds=1.79, P<0.05), disability of lower extremities (odds=1.63, P<0.05) and hearing impairment (odds=2.01, P<0.05) whereas it was negatively related with living in detached house (odds=0.41, P<0.05). It was found that accidents caused health (82.0%) and financial (38.2%) problems in elderly and also the disabilities in daily activities (66.3%). Average numbers of days with disability in daily activities were 21.1 in 1 year per accident and 5.7 in 1 year per individual. Occurrence of health problems and disability in daily activities were more frequent among women compared to men (P<0.05). Methods like environmental measures or educational programs to prevent accidents and accident-related injuries must be focused on defined risk groups and places where the accidents occur more frequently. Future researches about the effectiveness of prevention in elderly on accident frequency, mortality and morbidity are needed to deal with this current problem. PMID- 12888224 TI - Relationship between the number of remaining teeth and physical activity in community-dwelling elderly. AB - The relationship between oral health and general health in the elderly has been much studied. However, further studies focussing on the influence of oral health on the quality of life (QOL) among the elderly are required. The goal of this study was to evaluate the relationship between oral health and physical or cultural activities. Subjects were 101 community-dwelling elderly persons who were functionally independent (mean age 70.3). Oral health status was evaluated according to the number of remaining teeth and the number of functional teeth. Physical and cultural activities were evaluated from self-reported information. The relationship between oral health and physical or cultural activities was examined by logistic regression analysis. About 60% of subjects took part in cultural activities, and less than half actively exercised (leisure sports, 33.6%; travel, 42.6%). Persons with 20 or more remaining teeth were more active in leisure sports (Odds ratio (OR)=4.86, 95% confidence interval (CI)=1.34, 17.38) and travel (OR=5.42, 95% CI=1.63, 18.08) than those with fewer than 20 remaining teeth. These results suggest that the number of remaining teeth is associated with physical activity in elderly persons. PMID- 12888225 TI - Olfactory identification in elderly Greek people in relation to memory and attention measures. AB - The University of Pennsylvania Smell Identification Test (UPSIT) and the Cross Cultural Smell Identification Test (CC-SIT) were administered to nondemented Greek participants ranging in age from 49 to 88 years together with tests of verbal memory from the Wechsler Memory Scale-3rd Edition (WMS-III). The test scores of the sample administered the CC-SIT were compared with the test scores of the 12 analogous UPSIT items of the sample administered the UPSIT. The percent of individuals correctly identifying each of the odorants of the UPSIT and CC-SIT is reported, together with means and standard deviations (S.D.) of the total smell scores. UPSIT performance in both the full test and the 12 analogous items was associated with WMS-III Logical Memory I performance after accounting for the effects of age, education and gender. CC-SIT performance was associated with gender, score on the Beck Depression Inventory-II and Logical Memory I performance. The study shows that olfactory identification is associated with verbal memory in nondemented individuals after accounting for demographic variables. PMID- 12888226 TI - Treatment of chronic heart failure in long-term care facilities: implications of recent heart failure guidelines recommendations. AB - Use of angiotensin-converting enzyme inhibitor in heart failure with left ventricular systolic dysfunction has revolutionized management of heart failure and resulted in several national heart failure guidelines. However, studies have documented underutilization of these life saving drugs. Subsequently, results of other randomized clinical trials revealed the beneficial effects of angiotensin receptor blockers, beta-blockers, and spironolactone in heart failure with systolic dysfunction, which prompted several other guidelines. However, all these guidelines were primarily restricted to systolic dysfunction. Heart failure is as much a geriatric syndrome as it is a cardiac. Most heart failure patients are older adults. About half of the heart failure in older adults is not associated with left ventricular systolic dysfunction. Two recent heart failure guidelines, from the European Society of Cardiology and the American College of Cardiology/American Heart Association, published respectively in September and December 2001 included recommendations for patients with preserved left ventricular systolic function and other special populations, including older adults and those at end of life. The purpose of this article is to review the relevance of these two guidelines to heart failure patients living in long-term care facilities. As our population is aging, more heart failure patients are expected to live in long-term care facilities. Appropriate evaluation and management of these patients are essential to improve quality of heart failure care in the coming decades. PMID- 12888227 TI - Sleepiness and sleep in elderly persons with tinnitus. AB - The aim of this study was to evaluate the relation of tinnitus to sleep and daytime sleepiness (DS) in a group of elderly men and women. A questionnaire survey was undertaken among 10,216 elderly subjects in northern Sweden. The mean (+/-s.d.) ages of the men and women were 73.0+/-6.0 and 72.6+/-6.7 years, respectively. Tinnitus was reported by 14.9% of the men and 12.0% of the women. The relative number with tinnitus was not related to age in men, but increased with age in women. Poor sleep was reported by 14.4% of the men and 27.9% of the women. Among subjects with tinnitus poor sleep and frequent waking were more common in both sexes, while difficulties in falling asleep after awakening at night were reported more often by women. DS was more common in subjects with tinnitus and the frequency was even more increased in those with both tinnitus and poor sleep. There was no further increase in DS in men and women on sleep medication. Thus, DS in elderly persons with tinnitus may due both to tiring effect of the annoying sound itself but also the negative effect by tinnitus on sleep. PMID- 12888228 TI - Characterizing the elderly homeless: a 10-year study in Israel. AB - Since 1980s there is a noticeable change in the homeless population in Western countries. Some researchers titled the change as the "new" homeless that are distinctly different than the "skid row" population described in the past. Among the "new" homeless there are increasing numbers of elderly subjects. The elderly homeless are a fragile and vulnerable group that suffers from high rates of physical and mental problems as well as increased mortality. The aim of the present study was to characterize the "new" elderly homeless. Over a 10-year period each homeless 65 years of age and older was assessed by a psychiatrist. The project was undertaken in Tel-Aviv, Israel, and was feasible due to close co operation between the Municipal Welfare Department and mental health consultants. The Structured Clinical Interview for DSM-IV (SCID) was the main diagnostic tool. Ninety-eight subjects of 2567 homeless persons located and contacted were elderly (3.8%). Mean age for the elderly sub-group was 71.7+/-5.3 years and the great majority were male 94/98 (95.9%). In 44/98 (44.9%), a formal DSM-IV axis I psychiatric disorder was diagnosed, most common being dementia (15/44) and schizophrenia (15/44). A significant minority of 13/44 (29.5%) were diagnosed and treated prior to becoming homeless. Physical co-morbidity was found in nearly 2/3 subjects. Following intensive case-management by social workers, 35/44 (79.5%) subjects were successfully placed in permanent housing. The main conclusion is that the "new" elderly homeless are typically males suffering from frequent psychiatric morbidity and physical co-morbidity. Intensive case-management succeeded in achieving return to permanent housing in the majority of subjects. PMID- 12888229 TI - Comparison of two functional independence scales with a participation measure in post-stroke rehabilitation. AB - The objectives of the study were to compare the association and responsiveness of the functional autonomy measurement system (SMAF) and functional independence measure (FIM) as outcome measures addressing functional independence in stroke patients involved in an intensive rehabilitation program and to compare their relationships with a social participation measure after rehabilitation period. One hundred and thirty-two people who had a stroke were evaluated with the SMAF and FIM during the rehabilitation period (T1: admission; T2: discharge; n=132) and twice after discharge (T3=2 weeks; n=118; T4=6 months later; n=102). At T3 and T4, a participation measure, the assessment of life habits (LIFE-H), was added. The main findings are: (1) the total scores on the SMAF and FIM are strongly correlated together (r=0.93 to 0.95; p<0.001); 2) the responsiveness of both functional independence scales is similar even though the SMAF total score is more responsive to change than the FIM total score (standardized response mean: 1.20 vs. 0.97; p<0.01); (3) the SMAF and FIM are related similarly to the daily activities domain of the participation scale; and finally (4) the social roles domain of the participation scale is less related to the SMAF and MIF than the daily activities domain; however, the SMAF score is more related to the social roles domain than the MIF. Our results support the need to use supplementary measures, such as participation measure, that cover not only physical function but also the other domains of participation, such as interpersonal relationships and leisure, that can be disrupted following a stroke. PMID- 12888230 TI - Late life cognition among men: a life course perspective on psychosocial experience. AB - This report assesses associations between trajectories of psychosocial experience over the life course and cognitive function in late life men. Survey data from the National Longitudinal Study of Older Men, a community sample of 1835 United States men aged 45-59 years in 1966, are used to explore the cognitive effects of personality (locus of control) and emotion (affect) as these attributes evolve from middle age to late life. Locus of control is very stable over time, whereas negative and positive affect fluctuate. Inverse relationships are found between cognitive function and external locus of control, enduring negative affect, and the absence of positive affect. Levels of education also moderate these effects. Low educational attainment appears to intensify the risk for poor cognitive function associated with mixed internal-external locus of control and poor emotional state over time. The connection between education, usually completed early in the life course, and late life cognitive outcomes emphasizes the importance of promoting educational attainment among young people. Among older men with low educational attainment, the identification of mood disorders and personality attributes that negatively impact cognition may lead to the development of appropriate interventions. PMID- 12888253 TI - Studies on skin aging. Preparation and properties of fucose-rich oligo- and polysaccharides. Effect on fibroblast proliferation and survival. AB - Skin aging represents an important chapter of connective tissue aging and concerns an organ of vital importance. Here we describe the preparation as well as the biological properties of fucose-rich oligo- and polysaccharides (FROPs), composed of polymers of a trisaccharide containing galactose, acetyl galacturonic acid and fucose, from the original high molecular weight bacterial polysaccharide (Fucogel), Solabia, France). Using endoglycosidases, oligo- and polysaccharides were prepared and characterized by physical and chemical procedures. The non reducing end-groups comprise equal amounts of galactose and fucose. The here described biological properties are: stimulation of cell proliferation of cultured human skin fibroblasts, protection of cells against ascorbate-induced cytotoxicity due to the release of reactive oxygen species (ROS). Properties elsewhere described concern the inhibition of matrix metallo-proteinases 2 and 9 (MMP-2 and MMP-9), their expression and activation. Using fluorescein isothiocyanate (FITC)-labeled polysaccharides, their interaction with cell membranes and also their penetration and accumulation in cells, especially in the cell nucleus could be demonstrated, probably via cell-membrane receptor-mediated mechanisms. We describe some of the symptoms of skin aging and show, that the here-described polysaccharide preparations are susceptible to slow down some of the cellular and molecular mechanisms involved, partly by the mediation of the above-mentioned receptors, partly by acting directly on the regulation of gene expression. PMID- 12888254 TI - Extracellular matrix remodeling and matrix metalloproteinases in the vascular wall during aging and in pathological conditions. AB - The extracellular matrix provides a structural framework essential for the functional properties of vessel walls. The three dimensional organization of the extracellular matrix molecules - elastin, collagens, proteoglycans and structural glycoproteins - synthesized during fetal development - is optimal for these functions. In uninjured arteries and veins, some proteases are constitutively expressed, but through the control of their activation and/or their inhibition by inhibitors, these proteases have a very low activity and the turn-over of elastic and collagen fibers is low. During aging and during the occurrence of vascular pathologies, the balance between proteases and their inhibitors is temporally destroyed through the induction of matrix metalloproteinase gene expression, the activation of zymogens or the secretion of enzymes by inflammatory cells. Smooth muscle cells, the most numerous cells in vascular walls, have a high ability to respond to injury through their ability to synthesize extracellular matrix molecules and protease inhibitors. However, the three dimensional organization of the newly synthesized extracellular matrix is never functionally optimal. In some other pathologies - aneurysm - the injury overcomes the responsive capacity of smooth muscle cells and the quantity of extracellular matrix decreases. In conclusion, care should be taken to maintain the vascular extracellular matrix reserve and any therapeutic manipulation of the protease/inhibitor balance must be perfectly controlled, because an accumulation of abnormal extracellular matrix may have unforeseen adverse effects. PMID- 12888255 TI - The anti-ageing effects of caloric restriction may involve stimulation of macroautophagy and lysosomal degradation, and can be intensified pharmacologically. AB - Caloric restriction (CR) and a reduced growth hormone (GH)-insulin-like growth factor (IGF-1) axis are associated with an extension of lifespan across taxa. Evidence is reviewed showing that CR and reduced insulin of GH-IGF-1 axis may exhibit their effects at least partly by their common stimulatory action on autophagy, the cell repair mechanism responsible for the housekeeping of cell membranes and organelles including the free radical generators peroxisomes and mitochondria. It is shown that the life-long weekly administration of an anti lipolytic drug may decrease glucose and insulin levels and stimulate autophagy and intensify anti-ageing effects of submaximal CR. PMID- 12888256 TI - Effect of L-fucose and fucose-rich oligo- and polysaccharides (FROP-s) on skin aging: penetration, skin tissue production and fibrillogenesis. AB - Skin thickness is decreasing with age. This loss concerns both dermis and epidermis, cells and extracellular matrix. We could show here that percutaneous application of an L-fucose-containing preparation produced an increase of skin thickness and a densification of collagen bundles. We also could show that 3H-L fucose penetrates in the dermis, a prerequisite for the above mentioned favorable pharmacological activities. These results, together with the previous favorable activities on the downregulation of matrix-degrading enzymes, free radical scavenging and increased cell proliferation confirm the favorable action of fucose and fucose-rich polysaccharides (FROP-s) on the skin by slowing down its aging. PMID- 12888257 TI - The age-dependent vasodilatation and endothelial calcium influx induced by elastin peptides are modulated by extracellular glucose level. AB - Elastin peptides have been shown to produce many biological effects on various cell types, including an endothelium- and NO-dependent vasodilatation mediated by extracellular calcium influx and intracellular calcium elevation. Under normal concentration of extracellular glucose, the vasodilatory effect is observed in adult rats and is lost with age. Here, we have studied the consequences of extracellular glucose level changes on these effects triggered by elastin peptides (10(-4)-10(-3) mg ml(-1)), on 6- and 30-month-old rats, using the tension myography and the patch-clamp techniques. Our results show that low (0 mM) or high (33 mM) extracellular glucose concentrations abolish the extracellular calcium influx induced, under normal glucose level (11 mM), by the elastin peptides in cultured human endothelial cells. Also, low or high glucose abolish the vasodilatory action of elastin peptides observed on aorta rings from adult rats under normal glucose concentration. On the contrary, a dilation of aged rat aorta is observed in the presence of elastin peptides and high glucose, whereas such dilation is not observed when the elastin peptides are added in the presence of normal glucose concentration. In aging, a restoration by high glucose of the NO-dependent vasodilatation induced by elastin peptides could enhance the production of damaging peroxynitrite, potentially altering the structure and function of the blood vessels. These results could be of importance in the evaluation and treatment of aged patients with pathophysiological dysregulations of the circulating glucose level, such as in diabetes, age-related glucose intolerance, or low glucose levels caused by inappropriate glucose control treatments. PMID- 12888258 TI - Proteolyzed matrix as a template for the regulation of tumor progression. AB - Pericellular proteolysis plays a pivotal function in cell invasion, a hallmark of tumor growth and metastasis. The minidegradome constituted of two matrix metalloproteinases (MMP), i.e. MMP-2 and MT1-MMP, associated with tissue inhibitor of metalloprotease-2 (TIMP-2) and integrin (alpha(v)beta(3)) or CD(44), is mainly involved in such invasive program. It catalyzes matrix degradation but, alternatively, proteolytic exposure of matricryptic sites or matrikines liberation by those enzymes regulates either positively or negatively tumor cell migration. That applies to types I and IV collagens, elastin, laminin 5, as described here, but such phenomenon might be extended to other matrix macromolecules. The development of tumors from epithelium origin is related to aging. Senescent fibroblasts are characterized by increased expression of MMPs, (particularly collagenase-1 (MMP-1) and stromelysin-1 (MMP-3)) and deposited matrix by those aged cells was shown to favor cancer cell growth. Thus, compositional variation of matrix-surrounding tumor cells, with formation of matricryptic sites and matrikines, can be considered as one main epigenetic factor contributing to tumor progression. A matrix-directed pharmacological approach in cancer is now emerging. PMID- 12888259 TI - Drug therapy of diabetes in the elderly. AB - The prevalence of type 2 diabetes mellitus is increasing in the elderly. Although type 1 diabetic patients are living longer, over 95% of elderly people affected with diabetes have type 2 diabetes. This segment of the population, comprising men and women older than 65 years of age, is more likely to develop coexisting illnesses predisposing to the development of diabetes and complicating its management. In addition to changes in lifestyle, inherent age-related changes in carbohydrate metabolism contribute to the development of type 2 diabetes in old age. The long-term outcome of tight blood sugar control in the elderly is not known. Nevertheless the principles of management of type 2 diabetes in the elderly are essentially the same as in young- or middle-aged diabetic patients. Lifestyle modifications remain the cornerstone of medical therapy. When diet and exercise fail to achieve proper glycemic control, oral pharmacologic therapy and/or insulin therapy is indicated. The recently available oral glucose lowering agents in the market along with the newer types of insulin can be used in elderly diabetic patients. The effect of aging on metabolism and drug elimination kinetics must, however, be taken into consideration. In particular, it should be borne in mind that the risk of hypoglycemia is more deleterious in the elderly and should be avoided. In this review, the various pharmacologic agents available for the management of diabetes will be reviewed and some pertinent clinical guidelines will be suggested. PMID- 12888260 TI - Pharmacological properties of fucose. Applications in age-related modifications of connective tissues. AB - Fucose is the only component of glycoconjugates of vertebrates in the L configuration. It exhibits a number of unique and interesting biological properties reviewed briefly in this article. Its constant end-standing position on glycan chains predisposes fucose to play a key role in cell-cell and cell matrix interactions, mediated by several receptors such as those recognising the Lewis-type blood group substances, fucose-recognising lectines and the mannose fucose receptors. Some of the as yet unstudied or less well understood properties of L-fucose were explored in the present study, as its non-enzymatic interaction with amine-groups on macromolecules, its cellular uptake attributed to specific transport mechanisms and its effect on fibroblast cell cultures. We could document the stimulation of cell-proliferation and the inhibition of MMP expression and activation, both for MMP-2 and MMP-9. These and the other shortly reviewed properties of L-fucose may play an important role in its biological applications and actions. PMID- 12888261 TI - Immune dysfunction in the elderly and its reversal by antihistamines. AB - The decline in immunity seen in the elderly is a significant contributor to disease burden. This decline has largely been attributed to alterations in T cell immunity and contributes to an overall increased risk and severity of infection in the elderly. A key component of T cell immunity involves antigen presentation, an event where an antigen is processed and presented to specific immune cells for destruction. This event has been found to be crucial to immune function. Recent research has focused on a key antigen presenting cell (APC), the dendritic cell (DC), and changes within its function associated with aging. DCs are considered to be the most professional APCs, and are responsible for the initiation and outcome of effector T cells and their resultant immune response. DCs capture antigens and undergo a maturation process and polarize into either type 1 dendritic cells (DC1) or type 2 dendritic cells (DC2), based upon their ability to favor a T helper1 (Th1) or T helper 2 (Th2) T cell response, respectively. Evidence suggests that in normal healthy adults, a Th1 type response predominates, and in frail elders, a Th2 response predominates. It has been proposed that this change from a predominately Th1 type to a predominate Th2 type response is a possible mechanism for age-associated immune dysfunction. In addition, recent research has focused on how histamine, an inflammatory mediator, promotes a Th2 response. Histamine has also been shown to polarize human DCs into Th2 cell-promoting effector DCs or DC2s. This has been shown to occur via interaction with the H2 receptor. Therefore, we theorize that use of an H2 selective antihistamine will reverse this polarization back to a Th1 type response and therefore improve immune function of the frail elderly. PMID- 12888262 TI - Biochemical pharmacology of functional foods and prevention of chronic diseases of aging. AB - The major mechanisms and consequences of free radicals in foods and biological systems that result in the occurrence of chronic diseases of aging (atherosclerosis, cataract, cancers, diabetes, neurological diseases, immune inflammatory disorders) are described. The main antioxidant defense mechanisms from foods and cells and tissues are also reported. Many nutraceutical substances, with their respective beneficial actions (antioxidant, detoxifying, apoptotic actions) and effects, and most common food sources are also described. There are many options of foods to prevent cancer and chronic diseases, to improve life's quality in maturity. PMID- 12888263 TI - New perspectives in arsenic-induced cell signal transduction. AB - Although the carcinogenicity of arsenic has been well established, the underlying molecular mechanisms have not yet been fully identified. Accumulating evidence indicates that the alteration of cellular signal transduction is directly related to the carcinogenesis of arsenic. This review focuses on recent advances in arsenic-induced signal transduction, including reactive oxygen species (ROS) production, tyrosine phosphorylation, MAPK signaling, NF-kappaB activation, cell cycle arrest, and apoptosis. PMID- 12888264 TI - Functional interaction of cytochrome P450 with its redox partners: a critical assessment and update of the topology of predicted contact regions. AB - The problem of donor-acceptor recognition has been the most important and intriguing one in the area of P450 research. The present review outlines the topological background of electron-transfer complex formation, showing that the progress in collaborative investigations, combining physical techniques with chemical-modification and immunolocalization studies as well as site-directed mutagenesis experiments, has increasingly enabled the substantiation of hypothetical work resulting from homology modelling of P450s. Circumstantial analysis reveals the contact regions for redox proteins to cluster on the proximal face of P450s, constituting parts of the highly conserved, heme-binding core fold. However, more variable structural components located in the periphery of the hemoprotein molecules also participate in donor docking. The cross reactivity of electron carriers, purified from pro- and eukaryotic sources, with a diversity of P450 species points at a possible evolutionary conservation of common anchoring domains. While electrostatic mechanisms appear to dominate orientation toward each other of the redox partners to generate pre-collisional encounter complexes, hydrophobic forces are likely to foster electron transfer events by through-bonding or pi-stacking interactions. Moreover, electron tunneling pathways seem to be operative as well. The availability of new P450 crystal structures together with improved validation strategies will undoubtedly permit the production of increasingly satisfactory three-dimensional donor acceptor models serving to better understand the molecular principles governing functional association of the redox proteins. PMID- 12888265 TI - Synthesis, structural characterization and antimicrobial activities of 12 zinc(II) complexes with four thiosemicarbazone and two semicarbazone ligands. AB - Twelve zinc(II) complexes with thiosemicarbazone and semicarbazone ligands were prepared and characterized by elemental analysis, thermogravimetric and differential thermal analysis (TG/DTA), FT-IR and 1H and 13C NMR spectroscopy. Seven three-dimensional structures of zinc(II) complexes were determined by single-crystal X-ray analysis. Their antimicrobial activities were evaluated by MIC against four bacteria (B. subtilis, S. aureus, E. coli and P. aeruginosa), two yeasts (C. albicans and S. cerevisiae) and two molds (A. niger and P. citrinum). The 5- and 6-coordinate zinc(II) complexes with a tridentate thiosemicarbazone ligand (Hatsc), ([Zn(atsc)(OAc)](n) 1, [Zn(Hatsc)(2)](NO(3))(2).0.3H(2)O 2, [ZnCl(2)(Hatsc)] 3 and [Zn(SO(4))(Hatsc)(H(2)O)].H(2)O 4 [Hatsc=2-acetylpyridine(thiosemicarbazone)]), showed antimicrobial activities against test organisms, which were different from those of free ligands or the starting zinc(II) compounds. Especially, complex 2 showed effective activities against P. aeruginosa, C. albicans and moderate activities against S. cerevisiae and two molds. These facts are in contrast to the results that the 5- or 6-coordinate zinc(II) complexes with a tridentate 2 acetylpyridine-4N-morpholinethiosemicarbazone, ([Zn(mtsc)(2)].0.2EtOH 5, the previously reported catena-poly [Zn(mtsc)-mu-(OAc-O,O')](n) and [Zn(NO(3))(2)(Hmtsc)] [Hmtsc=2-acetylpyridine (4N-morpholyl thiosemicarbazone)]), showed no activities against the test microorganisms. The 5- and 6-coordinate zinc(II) complexes with a tridentate 2-acetylpyridinesemicarbazone, ([Zn(OAc)(2)(Hasc)] 6 and [Zn(Hasc)(2)](NO(3))(2) 7 [Hasc=2 acetylpyridine(semicarbazone)]), showed no antimicrobial activities against bacteria, yeasts and molds. Complex [ZnCl(2)(Hasc)] 8, which was isostructural to complex 3, showed modest activity against Gram-positive bacterium, B. subtilis. The 1:1 complexes of zinc(II) with pentadentate thiosemicarbazone ligands, ([Zn(dmtsc)](n) 9 and [Zn(datsc)](n) 10 [H(2)dmtsc=2,6-diacetylpyridine bis(4N morpholyl thiosemicarbazone) and H(2)datsc=2,6-diacetylpyridine bis(thiosemicarbazone)]), did not inhibit the growth of the test organisms. On the contrary, 7-coordinate zinc(II) complexes with one pentadentate semicarbazone ligand and two water molecules, ([Zn(H(2)dasc)(H(2)O)(2)](OAc)(2).5.3H(2)O 11 and [Zn(H(2)dasc)(H(2)O)(2)](NO(3))(2).H(2)O 12 [H(2)dasc=2,6-diacetylpyridine bis(semicarbazone)]), showed modest to moderate activities against bacteria. Based on the X-ray structures, the structure-activity correlation for the antimicrobial activities was elucidated. The zinc(II) complexes with 4N substituted ligands showed no antimicrobial activities. In contrast to the previously reported nickel(II) complexes, properties of the ligands such as the ability to form hydrogen bonding with a counter anion or hydrated water molecules or the less bulkiness of the 4N moiety would be a more important factor for antimicrobial activities than the coordination number of the metal ion for the zinc(II) complexes. PMID- 12888266 TI - New organotropic compounds. Synthesis, characterization and reactivity of Pt(II) and Au(III) complexes with bile acids: DNA interactions and 'in vitro' anticancer activity. AB - Based on the ability of bile acids to vectorialize the cytostatic activity of other agents, we have designed and synthesized a new series of platinum and gold complexes. These compounds were studied and characterized by elemental analysis, FT-IR, FAB(+)/MS, 1H, 13C and 195Pt NMR, UV-Vis spectroscopy and conductivity measurements in solution, among other techniques. Kinetic studies carried out in aqueous solution and in the presence of different NaCl concentrations: 4 mM (similar to cytoplasmic concentration), 150 mM (similar to plasmatic concentration). The effects on the electrophoretic mobility of the pUC18 plasmid, the DNA denaturation temperature, and ethidium bromide (EtBr) binding to DNA were studied. The complexes are able to inter-react with DNA to inhibit DNA synthesis and hence, to reduce cell proliferation. The complexes were evaluated for in vitro cytostatic activity against human colon adenocarcinoma, mouse hepatoma, human hepatoma, mouse leukemia, etc. The antitumor effect of some of the compounds prepared was similar to that of cisplatin. However, other compounds had lower cytostatic activity. This different behavior can be accounted for by the structure/activity relationship (SAR), although other factors, such as uptake and the different kinetic behavior in solution, may be responsible for these differences. PMID- 12888267 TI - Mechanism of insulin sensitization by BMOV (bis maltolato oxo vanadium); unliganded vanadium (VO4) as the active component. AB - Organovanadium compounds have been shown to be insulin sensitizers in vitro and in vivo. One potential biochemical mechanism for insulin sensitization by these compounds is that they inhibit protein tyrosine phosphatases (PTPs) that negatively regulate insulin receptor activation and signaling. In this study, bismaltolato oxovanadium (BMOV), a potent insulin sensitizer, was shown to be a reversible, competitive phosphatase inhibitor that inhibited phosphatase activity in cultured cells and enhanced insulin receptor activation in vivo. NMR and X-ray crystallographic studies of the interaction of BMOV with two different phosphatases, HCPTPA (human low molecular weight cytoplasmic protein tyrosine phosphatase) and PTP1B (protein tyrosine phosphatase 1B), demonstrated uncomplexed vanadium (VO(4)) in the active site. Taken together, these findings support phosphatase inhibition as a mechanism for insulin sensitization by BMOV and other organovanadium compounds and strongly suggest that uncomplexed vanadium is the active component of these compounds. PMID- 12888268 TI - The interaction of peptide-tethered platinum(II) complexes with DNA. AB - The sequence specificity and intensity of DNA damage induced by six peptide tethered platinum complexes was compared to cisplatin and Pt(en)Cl(2). DNA damage was investigated in pUC19 plasmid and in intact HeLa cells, and quantitatively analyzed using a Taq DNA polymerase/linear amplification assay. The DNA sequence specificity of the peptide-platinum compounds was found to be very similar to cisplatin and Pt(en)Cl(2), with runs of consecutive guanines being the most intensely damaged sites. The observed reactivity of the peptide-platinum complexes towards plasmid DNA was lower compared to cisplatin and Pt(en)Cl(2), with the glycine-tethered complex 3 and the phenylalanine-tethered complex 4 producing the highest relative damage intensity, followed by (in decreasing order) lysine-tethered (5), arginine-tethered (6), serine-tethered (7) and glutamate-tethered (8). The reactivity of the peptide-platinum complexes towards cellular DNA was also lower compared to cisplatin and Pt(en)Cl(2). For most investigated complexes, the relative damage intensities were found to be similar in cells compared to plasmid DNA, but were greatly reduced for 3 and 4. The lysine-tethered 5 complex produced the highest DNA damage intensity in cells followed by (in decreasing order) 6, 7, 3, 4 and 8. PMID- 12888269 TI - Synthesis and cytotoxicity of new platinum(IV) complexes of mixed carboxylates. AB - In order to develop new antitumor platinum(IV) complexes with highly tuned lipophilicity, a series of (diamine)Pt(IV) complexes of the formula [Pt(IV)(dach)L(3)L'] or [Pt(IV)(dach)L(2)L"(2)] (dach=trans-(+/-)-1,2 diaminocyclohexane; L=acetato, propionato; L'=acetato, propionato, valerato or pivalato; L"=trifluoroacetato) have been synthesized by electrophilic substitution of the tris(carboxylato)hydroxoplatinum(IV) complexes, [Pt(IV)(dach)L(3)OH] (L=acetato, propionato), with various carboxylic anhydrides such as acetic, trifluoroacetic, pivalic and valeric anhydrides. The present platinum(IV) complexes were fully characterized by means of elemental analyses, 1H NMR, mass and IR spectroscopies. The complexes 8 and 10, satisfying the appropriate range of lipophilicity (logP=0.18-1.54), exhibited high activity (ED(50), 5.1 and 1.3 microM, respectively) compared with other complexes, which implies that the lipophilicity is an important factor for the antitumor activity of this series of complexes. PMID- 12888270 TI - Interaction centres of purine nucleotides: adenosine-5'-diphosphate and adenosine 5'-triphosphate in their reactions with tetramines and Cu(II) ions. AB - The interactions between the nucleotides: adenosine-5'-diphosphate (ADP) and adenosine-5'-triphosphate (ATP) with spermine (Spm) and 1,11-diamine-4,8 diazaundecane (3,3,3-tet), as well as Cu(II) ions are studied. In the metal-free systems nucleotide-polyamine molecular complexes have been found to form, in which the interaction centres are the nitrogen atoms of the purine ring N(1) and N(7), oxygen atoms of the phosphate group of the nucleotide (for 3,3,3-tet) and protonated nitrogen atoms of the polyamine. Significant differences in the mode of metallation between the systems with Spm and 3,3,3-tet have been established. In the systems with Spm, the main products are protonated species with [N(7),O] chromophore and the nitrogen N(1) is involved in the intramolecular interaction additionally stabilising the complex. In the systems with 3,3,3-tet the formation of metal-ligand-ligand (MLL) species has been observed, in which the oxygen atoms from the phosphate group and the nitrogen atoms from the polyamine are involved in the metallation, while the N(1) and N(7) atoms from the purine ring of the nucleotide remain outside the inner coordination sphere of the copper ion. The main centre of metallation in the nucleotide, both with Spm and 3,3,3-tet, is the phosphate group of the nucleotide. PMID- 12888271 TI - Kinetic study of azole-bridged dinuclear platinum(II) complexes reacting with a hairpin-stabilized double-stranded oligonucleotide. AB - The cytotoxic dinuclear platinum(II) complexes [[cis-Pt(NH(3))(2)](2)(mu-OH)(mu pz)](NO(3))(2) (pz=pyrazolate) (1) and [[cis-Pt(NH(3))(2)](2)(mu-OH)(mu-1,2,3-ta N1,N2)](NO(3))(2) (1,2,3-ta=1,2,3-triazolate) (2), were allowed to react with the hairpin-stabilized double-stranded oligonucleotide d(TATGGCATT(4)ATGCCATA), to determine the amounts of intrastrand and interstrand DNA adducts. The reaction kinetics was investigated by reversed-phase HPLC, and the resulting products were analyzed using mass spectroscopy combined with enzymatic digestion, and Maxam Gilbert sequencing. The reaction of 1 results in the formation of the 1,2 intrastrand d(GG) adduct as the major final product. The two most abundant products of 2 were identified as isomeric 1,2-intrastrand d(GG) adducts differing probably in platinum coordination to the triazole ring. No GG-interstrand crosslinks were detected with either compound. d(GGC)-d(GCC) sequences of DNA do thus not appear to represent significant targets for forming interstrand crosslinks with either 1 or 2. PMID- 12888272 TI - Oxidative DNA damage of mixed copper(II) complexes with sulfonamides and 1,10 phenanthroline. Crystal structure of [Cu(N-quinolin-8-yl-p toluenesulfonamidate)2(1,10-phenanthroline)]. AB - Mixed coordination compounds of Cu(II) with sulfonamides and 1,10-phenanthroline as ligands have been prepared and characterised. Single crystal structural determination of the complex [Cu(N-quinolin-8-yl-p toluenesulfonamidate)(2)(phen)] shows Cu(II) ions are located in a highly distorted octahedral environment, probably as a consequence of the Jahn-Teller effect. The FT-IR and electronic paramagnetic resonance (EPR) spectra are also discussed. The mixed complexes prepared undergo an extensive DNA cleavage in the presence of ascorbate and hydrogen peroxide. Two of the complexes have higher nucleolytic efficiency than the bis(o-phenanthroline)copper(II) complex. PMID- 12888273 TI - Low-temperature (180 K) crystal structure, electron paramagnetic resonance spectroscopy, and propitious anticonvulsant activities of CuII2(aspirinate)4(DMF)2 and other CuII2(aspirinate)4 chelates. AB - The purpose of this research was to characterize by X-ray crystallography the ternary dimethylformamide (DMF) Cu(II) complex of acetylsalicylic acid (aspirin), in an effort to compare the structure-activity relationships for the anticonvulsant activity of this and other Cu(II)aspirinate chelates. The ternary DMF Cu(II) complex of aspirin was synthesized and crystals grown from a DMF solution were characterized by single crystal X-ray diffraction. This crystalline material was analyzed for anticonvulsant activity in the Maximal Electroshock (MES) Grand Mal and subcutaneous Metrazol (scMET) Petit Mal models of seizure used to detect anticonvulsant activity. The ternary DMF complex was found to be a monomolecular binuclear complex, tetrakis-mu (acetylsalicylato)bis(dimethylformamido)dicopper(II) [Cu(II)(2)(aspirinate)(4)(DMF)(2)] with the following parameters: monoclinic, space group P2(1)/n, a=12.259 (1), b=10.228 (1), c=16.987 (1) A, beta=92.07 (1) degrees; V=2128.5 (3) A(3); Z=2. The structure was determined at 180 K from 2903 unique reflections (I>1sigma(I)) to the final values of R=0.030 and wR=0.033 using F. This binuclear complex contains four acetylsalicylate bridging ligands which are related to each other in a two by two symmetry center. The four nearest O atoms around each Cu atom form a closely square planar arrangement with the square pyramidal coordination completed by the dimethylformamide oxygen atom occupying an apical position at a distance of 2.154 (1) A. Each Cu atom is displaced towards the DMF ligand by 0.187 A from the plane of the four O atoms. Electron paramagnetic resonance (EPR) spectra of [Cu(II)(2)(aspirinate)(4)(DMF)(2)] crystals show a strong antiferromagnetic coupling of the copper atoms, similar to that observed with other binuclear copper(II)salicylate compounds. Studies used to detect anticonvulsant activity revealed that [Cu(II)(2)(aspirinate)(4)(DMF)(2)] was an effective anticonvulsant in the MES model of seizure but ineffective against scMET-induced seizures. The monomolecular ternary binuclear [Cu(II)(2)(aspirinate)(4)(DMF)(2)] complex is more effective in inhibiting MES-induced seizures than other binuclear or mononuclear Cu(II) chelates of aspirin including: binuclear polymeric [Cu(II)(2)(aspirinate)(4)], [Cu(II)(2)(aspirinate)(4)(H(2)O)], which is anticipated to be less polymeric, and monomolecular ternary [Cu(II)(2)(aspirinate)(4)(DMSO)(2)] and [Cu(II)(aspirinate)(2)(Pyr)(2)]. These and other chelates appear to be more effective in the scMET model of seizure than [Cu(II)(2)(aspirinate)(4)(DMF)(2)]. These structure-activity relationships support the potential efficacy of Cu chelates of aspirin in treating epilepsies. PMID- 12888274 TI - Crystal structure of a covalent intermediate of endogenous human arylsulfatase A. AB - The structures of human arylsulfatase A crystals soaked in solutions containing 4 methylumbelliferyl phosphate and O-phospho-DL-tyrosine have been determined at 2.7- and 3.2-A resolution, respectively. The formylglycine in position 69, a residue crucial for catalytic activity, was unambiguously identified in both structures as forming a covalent bond to the phosphate moiety. A hydroxyl group is present at the Cbeta of residue 69 and the formation of one out of two possible stereomeric forms is strongly favoured. The structures confirm the importance of the gem-diol intermediate in the arylsulfatase's catalytic mechanism. The presence of an apparently stable covalent bond is consistent with the weak phosphatase activity observed for human arylsulfatase A. The structures of the complexes suggest that phosphate ions and phosphate esters inhibit arylsulfatase in non-covalent and covalent modes, respectively. The metal ion present in the active site of arylsulfatase A isolated from human placenta is Ca(2+) and not Mg(2+) as was found in the structure of the recombinant enzyme. PMID- 12888275 TI - Antifertility, antibacterial, antifungal and percent disease incidence aspects of macrocyclic complexes of manganese(II). AB - Macrocyclic complexes of Mn(II) were synthesized by template condensation using 2,6-diaminopyridine and diethylenetriamine with malonic, succinic, glutaric and adipic acids. The reaction proceeded smoothly to completion. These 16- to 24 membered N(6), but behaving as tetradentate, macrocyclic complexes were characterized by elemental analyses, molecular weight determinations, infrared, electronic, mass and X-ray spectral analyses. The elemental analyses are consistent with the formation of complexes [Mn(N(6)L(n))Cl(2)]. All the complexes are stable and monomeric in nature, as indicated by the molecular weight determinations. The spectral studies confirmed the proposed framework of the new macrocyclic complexes and indicated an octahedral geometry around the central metal atom. The complexes were screened in vitro against a number of pathogenic fungi and bacteria to assess their growth-inhibiting potential. The testicular sperm density, sperm morphology, sperm motility, density of cauda epididymis, spermatozoa and fertility in mating trials and the biochemical parameters of the reproductive organs of the rat were examined and are discussed. PMID- 12888276 TI - Enzyme-catalysed siloxane bond formation. AB - Biosilicification occurs on a globally vast scale under mild conditions. Although research has progressed in the area of silica biosynthesis, the molecular mechanisms of these interactions are effectively unknown. The natural production of silica in the Tethya aurantia marine sponge, Cylindrotheca fusiformis diatom, and Equisetum telmateia plant appear to be similar. However, the studies were complicated mechanistic queries due to the use of silicic acid analogues. Given these complications, a carefully chosen model study was carried out to test the ability of enzymes to catalyse the formation of molecules with a single siloxane bond during the in vitro hydrolysis and condensation of alkoxysilanes. Our data suggest that homologous lipase and protease enzymes catalyse the formation of siloxane bonds under mild conditions. Non-specific interactions with trypsin promoted the in vitro hydrolysis of alkoxysilanes, while the active site was determined to selectively catalyse the condensation of silanols. PMID- 12888277 TI - Influence of copper(II) and magnesium(II) ions on the ciprofloxacin binding to DNA. AB - The influence of magnesium(II) and copper(II) ions on the binding of ciprofloxacin to double stranded calf thymus DNA was studied by fluorescence emission spectroscopy, ultraviolet- and circular dichroism (CD) spectroscopy. The interaction of ciprofloxacin and copper(II) ions was followed by strong fluorescence quenching which was almost unaffected by the presence of DNA. On the other hand, only a slight decrease in fluorescence emission intensity, which was enhanced in the presence of DNA, was observed for ciprofloxacin interaction with magnesium(II) ions. Furthermore, magnesium(II) ions increase the thermal stability of the DNA, while, in the presence of ciprofloxacin, the degree of stabilisation is smaller. In contrast, copper(II) ions destabilise double helical DNA to heat, while ciprofloxacin slightly affects only the second transition of the biphasic melting curve of calf thymus DNA. Magnesium(II) ions at 25 degrees C induce conformational transitions of DNA at concentrations of 1.5 mM and 2.5 M, as monitored by CD. On the other hand copper(II) ions induce only one conformational transition, at a concentration of 12.7 microM. At higher concentrations of copper(II) ions (c>700 microM) DNA starts to precipitate. Significant changes in the CD spectra of DNA were observed after addition of ciprofloxacin to a solution containing DNA and copper(II) ions, but not to DNA and magnesium(II) ions. Based on our spectroscopic results, we propose that copper(II) ions are not directly involved into ciprofloxacin binding to DNA via phosphate groups as it has been suggested for magnesium(II) ions. PMID- 12888278 TI - Spectral studies on aluminum ion binding to the ligands with phenolic group(s): implications for the differences between N- and C-terminal binding sites of human serum apotransferrin. AB - Both the binding and releasing of ferric ions in C-, and N-terminal binding sites of human serum transferrin are different. To understand the difference here the interactions of aluminum with the ligands containing phenolic group(s), including 8-hydroxyquinoline, salicylic acid, N,N'-di(2-hydroxybenzyl)ethylenediamine-N,N' diacetic acid, N,N'-ethylenebis[2-(o-hydroxyphenolic)glycine], and human serum apotransferrin, respectively, are investigated by using UV difference and fluorescence spectra methods in 0.1 M N-2-hydroxyethylpiperazine-N-2 ethanesulfonic acid at pH 7.4. Aluminum binding produces a UV difference peak near 235 nm that is characteristic of phenolic groups binding to aluminum. The peak at 235 nm has been used to determine conditional binding constants of log K(Al-HBED)=8.88+/-0.74 and log K(Al-EHPG)=9.38+/-0.03. However, the effects of aluminum binding on the fluorescence intensity of N,N'-ethylenebis[2-(o hydroxyphenolic)glycine], salicylic acid and N,N'-di(2-hydroxybenzyl) ethylenediamine-N,N'-diacetic acid, 8-hydroxyquinoline are disparate, the former showing a decrease and the latter an increase. At pH 7.4, there is N cdots, three dots, centered H-O type intramolecular hydrogen bond in 8-hydroxyquinoline, N,N' di(2-hydroxybenzyl) ethylenediamine-N,N'-diacetic acid and O cdots, three dots, centered H-O type intramolecular hydrogen bond in salicylic acid, N,N' ethylenebis[2-(o-hydroxyphenolic)glycine]. The effects of salts on the fluorescence intensity of the ligands containing phenolic group(s) show that fluorescence emission increases with the breaking of an N cdots, three dots, centered H-O type intramolecular hydrogen bond and fluorescence emission decreases with the breaking of an O cdots, three dots, centered H-O type intramolecular hydrogen bond. Fluorescence titrations of apotransferrin and both forms of monoferric transferrin with aluminum indicated that there is O cdots, three dots, centered H-O type intramolecular hydrogen bonds for the phenolic groups of Tyr426 and Tyr517 in the C-terminal binding site. While N cdots, three dots, centered H-O type intramolecular hydrogen bonds are found for the phenolic groups of Tyr95 and Tyr188 in the N-terminal binding site. PMID- 12888279 TI - Synthesis, structural characterization and in vitro cytotoxicity of organotin(IV) derivatives of heterocyclic thioamides, 2-mercaptobenzothiazole, 5-chloro-2 mercaptobenzothiazole, 3-methyl-2-mercaptobenzothiazole and 2-mercaptonicotinic acid. AB - Five new organotin(IV) molecules with the heterocyclic thioamides; 2 mercaptobenzothiazole (Hmbzt), 5-chloro-2-mercaptobenzothiazole (Hcmbzt), 3 methyl-2-mercaptobenzothiazole (mmbzt) and 2-mercaptonicotinic acid (H(2)mna) of formulae [(n-C(4)H(9))(2)Sn(mbzt)(2)] (1), [(C(6)H(5))(2)Sn(mbzt)(2)] (2), [(CH(3))(2)Sn(cmbzt)(2)].1.7(H(2)O)] (3), [(n C(4)H(9))(2)SnCl(2)(mmbzt)(2).(CH(2)Cl(2))] (4) and [[(C(6)H(5))(3)Sn](2)(mna).[(CH(3))(2)CO]] (5) have been synthesized and characterized by elemental analysis, 1H-, 13C-NMR, FT-IR and Mossbauer spectroscopic techniques. Crystal structures of molecules 1, 3 and 5 have been determined by X-ray diffraction at 173(1) K (1 and 5) and 293(2) K (3). Compound 1 C(22)H(26)N(2)S(4)Sn, is monoclinic, space group C2/c, a=44.018(2), b=8.8864(5), c=12.8633(7) A, beta=104.195(5) degrees, Z=8. Compound 3 is also monoclinic, space group P2(1)/c and a=17.128(2) A, b=17.919(2) A, c=7.3580(10) A, beta=98.290(10) degrees, Z=4. In both molecules 1 and 3, two carbon atoms from aryl groups, two sulfur and two nitrogen atoms from thione ligands form a distorted octahedral geometry around tin(IV) with trans-C(2), cis-N(2), cis-S(2) configurations. Compound 5 C(45)H(39)NO(3)SSn(2) is monoclinic, space group P2(1)/n, a=9.1148(2) A, b=29.2819(6), c=15.5556(4) A, beta=106.2851(9) degrees, Z=4. Complex 5 contains two [(C(6)H(5))(3)Sn(IV)] moieties linked by a double deprotonated 2-mercaptonicotinic acid (H(2)mna). Both tin(IV) ions are five coordinated. This complex is the an example of a pentacoordinated Ph(3)SnXY system with an axial-equatorial arrangement of the phenyl groups at Sn(1) atom. Compounds 1, 3 and 5 were tested for in vitro cytotoxicity against the cancer cell line of sarcoma cells (mesenchymal tissue) from the Wistar rat, polycyclic aromatic hydrocarbons (benzo[a]pyrene) carcinogenesis. Compound 5 exhibits strong cytotoxic activity, while complexes 1 and 3 show less cytotoxic activity. PMID- 12888281 TI - Commentary: the importance of demonstrations in developing adolescent HIV care models. PMID- 12888280 TI - Human ZIP1 is a major zinc uptake transporter for the accumulation of zinc in prostate cells. AB - The prostate gland of humans and other animals accumulates a level of zinc that is 3-10 times greater than that found in other tissues. Associated with this ability to accumulate zinc is a rapid zinc uptake process in human prostate cells, which we previously identified as the hZIP1 zinc transporter. We now provide additional evidence that hZIP1 is an important operational transporter that allows for the transport and accumulation of zinc. The studies reveal that hZIP1 (SLC39A1) but not hZIP2 (SLC39A2) is expressed in the zinc-accumulating human prostate cell lines, LNCaP and PC-3. Transfected PC-3 cells that overexpress hZIP1 exhibit increased uptake and accumulation of zinc. The V(max) for zinc uptake was increased with no change in K(m). Along with the increased intracellular accumulation of zinc, the overexpression of hZIP1 also results in the inhibition of growth of PC-3 cells. Down-regulation of hZIP1 by treatment of PC-3 cells with hZIP1 antisense oligonucleotide resulted in a decreased zinc uptake. Uptake of zinc from zinc chelated with citrate was as rapid as from free zinc ions; however, the cells did not take up zinc chelated with EDTA. The cellular uptake of zinc is not dependent upon an available pool of free Zn(2+) ions. Instead, the mechanism of transport appears to involve the transport of zinc from low molecular weight ligands that exist in circulation as relatively loosely bound complexes with zinc. PMID- 12888282 TI - Introduction: what youth need--adapting HIV care models to meet the lifestyles and special needs of adolescents and young adults. PMID- 12888283 TI - Case finding for HIV-positive youth: a special type of hidden population. AB - PURPOSE: To describe the HIV case finding strategies used by the Special Projects of National Significance (SPNS), Adolescent HIV Outreach and Treatment programs, the populations of youth they were able to reach, and the populations of HIV positive youth they were able to identify. METHODS: Program specifications from five programs located in four major metropolitan centers were contrasted. Four of the programs also provided outcome data for HIV counseling and testing outcome numbers, demographic and risk profile data for youth who underwent HIV testing, and mode of infection of HIV-positive youth. RESULTS: The program outcomes were discussed in terms of similarities and differences in outreach methods (e.g., peer workers, time of outreach, etc.), geographic settings (i.e., mobile van, institutional settings, community locations), individual characteristics (e.g., pregnant women) and youth subcultures (i.e., gay/transgendered, incarcerated juveniles, homeless). CONCLUSIONS: Because HIV-positive adolescents will constitutionally remain a "hidden population," a great deal of time and effort will continue to need to go into the front end of outreach, counseling and testing. Specific guidance and recommendations for locating HIV-positive youth were provided to program designers for each type of outreach strategy. PMID- 12888284 TI - Transitioning youths into care: linking identified HIV-infected youth at outreach sites in the community to hospital-based clinics and or community-based health centers. AB - PURPOSE: To describe and analyze the process of transitioning HIV-infected youths from the point of HIV diagnosis into HIV treatment. Rather than simply offering HIV positive youths a list of referrals, the youth-focused SPNS grantees (AWAC) found it vital that youths were immediately assisted with linkage to a medical provider. METHODS: From February 1997 to December 2000, 107 identified HIV infected youths from the five adolescent SPNS projects were surveyed on needs and barriers. The time interval between HIV testing and youth linkage to care was also noted. RESULTS: Nine percent of youth reported perceived barriers to accessing health care. Perceived needs were identified as Mental Health (44.9%; n = 48); Alcohol and drug treatment (14%; n = 15); transportation to health care settings (40.2%; n = 43); and housing (46.7%; N = 50). At sites tracking linkage to care, the time of being transitioned into a medical setting ranged from 5 to 55 days (average 26 days). CONCLUSIONS: The period of transitioning identified HIV-infected youths into care can be reduced from 1-5 years to as short as 5-55 days. Success with linking these youth to care involves establishing a series of contacts at outreach sites wherein program staff seeks to build trusting relationships with youths, is able to track these youths and identify and address perceived needs. PMID- 12888285 TI - The utilization of treatment and case management services by HIV-infected youth. AB - PURPOSE: This article describes the essential components for effective and comprehensive HIV care for youth who have tested positive and have been linked to HIV treatment. Descriptive profile data are also presented that detail the demographics, risk behaviors and health care barriers of youth served in the five Special Projects of National Significance (SPNS), which focused on adolescents and young adults. METHODS: Data presented are from the core multi-site data set, which was standardized across the five youth-oriented SPNS projects. Substance use and mental health symptoms were gathered using the Personal Problem Questionnaire (PPQ) screener, which was an adaptation of the PRIME-MD. In-depth qualitative interviews with enrolled HIV-positive youth were also conducted by several Projects. RESULTS AND CONCLUSIONS: Medical care alone is not enough and cannot be effective without supportive program components such as flexible scheduling, and a multi-disciplinary team approach that includes assertive case management. Case Managers help enrolled youth with concrete service needs such as housing, emergency financial assistance for food/utilities, transportation, child care, coverage for prescriptions, and public entitlements. They also help isolated youth to connect with a personal support system. Addressing those needs helps to facilitate and reinforce treatment adherence and retention. In addition to other identified needs such as stable housing and transportation, a significant number of enrolled youth self-reported having experienced physical, sexual, and/or emotional abuse in their lives and articulated a need for mental health services. Therefore, effective HIV care for youth must be multi-faceted; it must consist of more than a medical component. PMID- 12888286 TI - Retention, adherence, and compliance: special needs of HIV-infected adolescent girls and young women. AB - PURPOSE: To demonstrate that whereas all HIV-infected youth evidence complex factors that challenge retention in care and adherence to treatment, HIV-infected females have additional issues that are gender-specific. METHODS: Preliminary data from a subset of 21 adolescent/young women under age 25 from the Whole Life mental health-perinatal HIV care project were analyzed to illustrate the needs of these patients. RESULTS: Of the 21 young women assessed, all but one was of minority background, and a sizeable majority had limited education ( 0.05) by AFB(1). Egg weight was significantly lower ( p<0.05) only for groups exposed to 100 microg AFB(1) kg(-1). Residues of aflatoxins were detected in eggs at levels that ranged from 0.01 to 0.08 microg kg(-1) (AFB(1)), 0.03-0.37 microg kg(-1) (AFM(1)), 0.01-1.03 microg kg( 1)(AFB(2a)) and 0.01-0.03 microg kg(-1) (AFL). Results indicate that the excretion of aflatoxin residues in quail eggs might occur at relatively low concentrations under conditions of long-term exposure of quail to low levels of AFB(1). PMID- 12888391 TI - Occurrence of aflatoxins in layer feed and corn samples in Konya province, Turkey. AB - The natural occurrence of aflatoxin was investigated in layer feed and corn samples brought to Konya Veterinary Control and Research Institute Laboratory between 15 April and 15 December 2002. Seventy-eight samples (52 feeds, 26 corn samples) were analysed for total aflatoxin (B1 + B2 + G1 + G2) by an ELISA screening method. Aflatoxin contamination was deter-mined in 37 feed samples (71.1%) and 15 corn samples (57.7%), with a range of 1.5-133 microg kg(-1). However, a majority of the aflatoxin contamination was less than 5 microg kg(-1) (50% within the positive samples). Two feed samples and two corn samples exceeded the maximum tolerated levels in feed (20 microg kg(-1)) and feedstuffs (50 microg kg(-1)) for total flatoxin. PMID- 12888392 TI - Congener-specific distribution of polychlorinated dibenzo-p-dioxins, dibenzofurans and biphenyls in animal feed. AB - Polychlorinated dibenzo-p-dioxins (PCDDs), dibenzofurans (PCDFs) and polychlorinated biphenyls (PCBs) were extracted by accelerated solvent extraction from animal feed samples and analysed by high-resolution gas chromatography/high resolution mass spectrometry (HRGC/HRMS). In all of the feed samples analysed, the concentrations of PCDD/Fs and PCBs were below the existing tolerance limit of 5 pg I-TEQ g(-1) and 200 ng g(-1) fat, respectively. The mean concentrations of the total PCDDs, total PCDFs and total PCBs were 1.73-11.50 pg g(-1), 0.23-11.91 pg g(-1) and 60.00-234.40 pg g(-1) feed, respectively. Investigation of the correlations among the concentrations of PCDDs, PCDFs and PCBs showed that samples containing increased amounts of PCDFs also contained higher concentrations of PCDDs, with an average ratio of PCDF:PCDD of about 1.23:1 (r(2)=0.72, p<0.05). The correlation between the concentrations of PCDD/Fs and PCBs was positive but not significant. PMID- 12888393 TI - Evaluation of some screening methods for the analysis of contaminants in recycled polyethylene terephthalate flakes. AB - A range of different analytical techniques were used to test recycled polyethylene terephthalate (PET) flakes for potential chemical contaminants. The techniques used were headspace gas chromatography coupled to mass spectrometry (GC-MS), liquid extraction followed by GC-MS, supercritical-fluid extraction followed by GC-MS, and migration testing followed by elemental analysis using inductively coupled plasma mass spectroscopy (ICP-MS). The PET samples were 50 representative samples taken from 600 that had been collected throughout Europe and which had been screened for potential contaminants using a single technique of high-temperature GC-MS. Six of the 50 samples tested had been spiked with a selection of model contaminants, three samples were virgin PET flakes and two of the samples were from supercleaning processes. All samples were analysed 'blind' in this exercise. The qualitative results showed that most of the contaminants came from the first use, being flavour-aroma compounds from soft drinks. The quantitative analysis found concentrations under a few mg kg(-1) in the polymer, except for the spiked samples. Element migrations were low and only calcium, silicon and sodium had median migrations above 50 microg l(-1). This in-depth analysis of recycled PET flakes did not identify any significant contaminants that had not already been detected by the high-temperature static GC-MS screening method, thus demonstrating its utility. PMID- 12888394 TI - Analytical methods for food-contact materials additives in olive oil simulant at sub-mg kg-1 level. AB - Methods of analysis for four additives (two antioxidants, IRGANOX 245 and 1035; an ultraviolet absorber, CHIMMASORB 81; and an optical brightening agent, UVITEX OB) in olive oil are reported. These additives have the potential to migrate from food-contact materials into the European Union fatty food simulant olive oil, which is the most difficult matrix for analysis. The additives were chosen because they differed in their chemically active groups, had different functions within the polymer, have low proposed specific migration limits and are commonly used in food-contact materials such as polystyrenes and polyolefins. The proposed analytical methods for the additives are simple, rapid, inexpensive and also broadly applicable to the aqueous food simulants. All methods were evaluated by constructing calibration curves, measurement of recovery and precision, and determining the limits of detection. Most of the methods involve direct injection of an olive oil solution for high-performance liquid chromatography analysis with ultraviolet-visible or fluorescence detection. The methods allowed establishment of additive stability and measurement of migration of the selected additives into olive oil at different time-temperature conditions used in migration studies into food simulants. PMID- 12888395 TI - Increased migration levels of bisphenol A from polycarbonate baby bottles after dishwashing, boiling and brushing. AB - Baby bottles are often made of polycarbonate plastic. Impurities remaining in the bottle from the monomer bisphenol A can migrate from the plastic bottles into baby food, thereby causing a health concern. Previous migration testing of new baby bottles showed only trace migration levels of the substance. In the present work, polycarbonate baby bottles were subjected to simulated use by dishwashing, boiling and brushing. Migration testing performed with both new and used bottles revealed a significant increase in migration of bisphenol A due to use. This finding might be explained by polymer degradation. Bisphenol A was determined in 200-ml samples of water food simulant by a method based on solid-phase extraction followed by gas chromatography coupled with mass spectrometry. The detection limit was 0.1 microg l(-1). Twelve different polycarbonate baby bottles were tested by filling them with hot water (100 degrees C) for 1 h. The mean bisphenol A level from new bottles was 0.23 + -0.12 microg l(-1), while the mean levels from bottles subjected to simulated use were 8.4 + -4 microg l(-1) (dishwashed 51 times) and 6.7 + -4 microg l(-1) (dishwashed 169 times), respectively. None of the bottles released bisphenol A at levels that exceed the recently established provisional tolerable daily intake (0.01 mg kg(-1) body weight/day) in the European Union. PMID- 12888397 TI - Self-assessment of DSM-IV criteria for major depression in psychiatric out- and inpatients. AB - The Depression Self-Rating Scale (DSRS) is a self-report inventory based on the DSM-IV criteria for major depression. This study aimed to investigate diagnostic concordance of a major depressive disorder (MDD) using the DSRS and the Structured Clinical Interview for DSM-IV, axis I disorders (SCID-I). An additional aim was to investigate whether concordance could be improved by adding different cut-off scores for the Beck Depression Inventory (BDI) or the self report version of the Montgomery-Asberg Depression Rating Scale (MADRS-S). MDD was determined by means of the SCID-I interview and the DSRS in 83 psychiatric in and outpatients. Depressive severity was assessed using the BDI and the MADRS-S. A DSRS-version including the A- and C-criteria for an MDD had a sensitivity and a specificity for an expert-rated diagnosis of 86% and 75%, respectively. By using cut-off scores for the BDI or the MADRS-S, specificity could be raised to 85% without loss of sensitivity. The DSRS appears to be a useful instrument in the diagnostic process involving MDD, particularly when used in combination with the BDI or the MADRS-S. The DSRS is thought to be of value in psychiatric outpatient departments, where treatment of patients is often delegated to paramedical staff or nurses. The scale might also be used as an independent diagnostic tool in epidemiological studies in order to achieve an estimate of the prevalence of MDD in different population settings. PMID- 12888398 TI - Evidence-based medicine, randomized controlled trials, publication bias, patient population, follow-up and access. PMID- 12888399 TI - Schizophrenia and contact with health and social services: a Nordic multi-centre study. AB - BACKGROUND: In a Nordic multi-centre study investigating the life and care situation of persons with schizophrenia living in the community, factors explaining use of health and social services were examined. METHOD: Four hundred and eighteen individuals with schizophrenia from 10 sites were interviewed about their contact with different services (support functions within and outside the mental health services, general practitioners (GPs), physicians in the mental health, psychotherapy, day-care and inpatient treatment), psychopathology, social network and needs for care. RESULTS: Physicians and support contacts within the mental health system were most used and GPs and psychotherapy least. Three groups of variables were stabile predictors of contact: rural-urban differences, diagnoses (hebephrenic schizophrenia associated with less contact with physicians in the mental services and more with GPs) and health needs as experienced by the patients. No differences between the centres with regard to total service use were found, but the patterns of contact reflected urban-rural variance. A low number of health needs predicted contact with physicians within the mental health services, whereas a high number of such needs was related to contact with GPs and support functions within the mental health services. Social relations exhibited the highest number of unmet needs. CONCLUSIONS: Contact with physicians working in the mental health services was much more common than contact with GPs. Based on a broad spectre of demographic, clinical and network variables, it was not possible to find models that explained substantial parts of the variance of service use. Patterns of contact were different in rural, town and city surroundings, and with the exception of psychotherapy, the rural pattern was characterized by use of less specialized services. The importance of health needs and diagnosis as predictors of contact illustrate the profound and lasting effects on health of having a diagnosis of schizophrenia. PMID- 12888400 TI - Dosage patterns of antipsychotic drugs for the treatment of schizophrenia in Swedish ambulatory clinical practice--a highly individualized therapy. AB - OBJECTIVE: To analyse the dosage pattern of antipsychotic drugs for schizophrenia in ambulatory care in Sweden. METHOD: The study was based on a nationwide consecutive weekly random sample of physicians during the period 1991-98 comprising 265,331 visits. RESULTS: In 515 visits (0.19%), antipsychotics were prescribed for schizophrenia. More than one antipsychotic (50 different combinations) was prescribed in 20% of the visits. Patients received higher total daily doses when prescribed more than one antipsychotic drug; mean dose in monotherapy was 210.7 chlorpromazine equivalents (CPZeq) and in polytherapy 406.8 CPZeq. CONCLUSION: Antipsychotics, in contrast to current recommendations, were prescribed as highly individualized therapies in a wide variety of doses and with a high frequency of polypharmacy. The combinations used are often unsuitable and may lead to unnecessary adverse effects. PMID- 12888401 TI - Educational intervention for the relatives of schizophrenia patients in Finland. AB - Two methods of providing an educational intervention for families of patients with schizophrenia in Finland were compared. The aims of the intervention were to improve relatives' level of knowledge about the illness, and change the level of expressed emotion (EE), objective burden and psychological distress. Sixty-nine persons participated in the oral presentation groups comprising eight sessions and 128 persons participated in the video education comprising six sessions. The education led to significant knowledge gains and to an increase in the psychological well-being of the participants, but there were no significant changes in objective burden or EE status after intervention. Furthermore, there was only one difference between the two methods of information delivery. The participants in the video groups felt more often that the lessons were useful to them than did the participants in the oral presentation groups. The evidence suggests that brief educational intervention can yield significant benefits in meeting the needs of family members. Better designed, randomized studies investigating the efficacy of brief educational interventions are needed. PMID- 12888402 TI - Test-retest reliability of the Camberwell Assessment of Need (CAN). AB - The Camberwell Assessment of Need (the CAN) is a widely used instrument in several countries to assess needs for the severely mentally ill. The instrument consists of ratings of needs made by patient and staff. This paper reports test retest reliability for staff ratings in the Swedish version. Fifty-six patients, most of them with a diagnosis of schizophrenia, were interviewed by one person from the social services and by one from the psychiatric clinic. Afterwards the ratings were discussed and negotiated in order to arrive at one unanimous rating. The same procedure was replicated after 2-4 weeks. The intraclass correlation coefficient for the sum of needs was 0.93, and for the sum of unmet needs and met needs 0.80 for both. There were no means differences between the first and second rating. An analysis was also made of the test-retest scores of individual needs and subscales. The results in this study suggest that the summary scores of the CAN generally are reliable over time when using the method applied here. General conclusions about test-retest reliability for individual needs and subscales are more difficult to make and demand larger samples. PMID- 12888403 TI - Health of the Nation Outcome Scales (HoNOS): Implementability, subscale structure and responsiveness in the daily psychiatric hospital routine over the first 18 months. AB - The Health of the Nation Outcome Scales (HoNOS) includes 12 items designed to measure the problems patients have when admitted to a psychiatric hospital and to monitor to what extent these problems have diminished at discharge from hospital. The Danish version of the HoNOS was translated from the original English version in accordance with the WHO guidelines for scale translation. The HoNOS was evaluated for inter-staff agreement using a case vignette from the English training sessions. The HoNOS was then applied in a consecutive sample of 1769 inpatients over a period of 18 months. This sample covered 89% of all admissions in this period and was found representative with regard to age and gender. The diagnostic distribution according to ICD-10 showed that half of the patients were listed within schizophrenia and mood disorders. The HoNOS profile, using all 12 items of the scale, showed that schizophrenia was associated with hallucinations or delusional symptoms, with social and cognitive problems; mania with aggressive behaviour, with drinking or drug-taking and with anxiety; whereas depression was associated with suicidal behaviour, physical illness and depressed mood. Long term stay in hospital was associated with a higher level of psychopathological symptoms at admission for schizophrenia, mania and depression. Among the diagnostic groupings, mania was associated with the greatest improvement in behavioural problems and psychological symptoms. The use of lithium was diagnostically associated with bipolar and schizoaffective disorders. Within these categories, the lithium-treated patients had a greater improvement in behavioural problems and psychological symptoms than the non-lithium-treated. The use of electroconvulsive therapy (ECT) in depressive patients was associated with a HoNOS admission profile of higher scores on psychological problems (delusions and depressed mood) than those seen in the non-ECT-treated depressive patients. The ECT-treated patients had a higher level of improvement at discharge. In conclusion, this first Scandinavian study with HoNOS showed that the scale is easy to implement in the daily routine of a psychiatric ward and that the HoNOS can give useful information about the quality of care in hospitalized patients. PMID- 12888404 TI - Measurement of depression in patients with chronic obstructive pulmonary disease (COPD). AB - OBJECTIVE: To estimate the validity of the Hamilton Depression Scale (HDS) in a population of patients with chronic obstructive pulmonary disease (COPD). METHODS: Forty-nine patients with moderate to severe COPD were examined using the ICD-10 criteria for depression. The mean age of the patients was 71 years and 33 (64%) were women. Forty-six (94%) of the patients were also evaluated using the 17-item HDS including the six-item Hamilton Depression subscale (HDSS). Internal and external validity were measured using factor analysis, Cronbach Coefficient alpha, Loevinger coefficient of homogeneity, correlation analysis and ROC-curves. RESULTS: Twenty-three (47%) of the patients were depressed according to the ICD 10 criteria for depression. The HDSS but not the HDS showed a good internal validity. An acceptable external validity was furthermore shown for the HDSS. CONCLUSION: The HDSS can be recommended as a suitable depression rating scale for COPD patients. PMID- 12888405 TI - Negative memories of childhood and current drug use. AB - Data on drug abuse and memories of the childhood were collected through a self report questionnaire from a group of current drug users and a group of non-using controls. Both samples were unidentified as groups by the society and were identified by the researchers throw snowball sampling. Earlier results of an unstable childhood and a poor social situation from studies that used other sampling methods were replicated. The drug users had an earlier nicotine and alcohol debut, and perceived themselves as unloved, physically abused children that were afraid of their parents during childhood. In addition, depression, suicide attempts and convictions were more common among the drug users. PMID- 12888406 TI - Self-image of adolescents with diabetes mellitus type-I and rheumatoid arthritis. AB - The purpose of this research was to determine possible differences in the self concept of chronically ill and healthy adolescents. A group of adolescents with diabetes mellitus type-I (DM) and a group with rheumatoid arthritis (RA) were chosen for the study, together with a control group without any chronic illness. The groups were matched for gender, age (DM 17.8, RA 17.9, control group 17.6 years) and social class. The Offer Self-Image Questionnaire (OSIQ) was used to estimate differences between the groups of 23 DM adolescents, 25 RA adolescents and 26 control group adolescents. The scores are reversed; thus the higher the score, the better the self-image. The results indicate that Body Image, and Vocational and Educational Goals were lower in the groups of chronically ill adolescents than in the control group; however, no statistically significant differences were found between the groups on the OSIQ scales. The self-image of adolescents with DM with good metabolic control and moderate and long-lasting RA is relatively well developed. PMID- 12888407 TI - Phenelzine efficacy in refractory social anxiety disorder: a case series. AB - Several effective treatments for social phobia are now available, notably cognitive behavioural therapy and some antidepressant medications. Controlled trials indicate that 25-45% of patients fail to respond to the initial treatment. To my knowledge, nothing has yet been published on the treatment of refractory cases. Phenelzine is effective in social phobia, and no treatment has been shown to be superior to phenelzine. Hence, phenelzine is a reasonable option for patients unresponsive to other treatments. This report presents the results of naturalistic, open-label phenelzine treatment of a series of seven consecutive, previously unresponsive patients treated with a mean dose of phenelzine 66 mg. Six patients were classified as responders according to Clinical Global Impression (CGI) ratings. Some experienced dramatic improvement and regained ability to function in school or work. Two patients experienced re-emergence of social phobia while on maintenance therapy. The majority of patients had a sustained response and elected to stay on the medication. There were no serious adverse events. Two patients discontinued due to side-effects. The drawbacks of phenelzine therapy may be more than outweighed by its potential utility in relieving social anxiety disorder. Phenelzine appears to be effective in treating social phobia that has been unresponsive to other treatments. No patient should be considered treatment resistant without being offered a trial of phenelzine. PMID- 12888408 TI - Gambling and problematic gambling with money among Norwegian youth (12-18 years). AB - An epidemiological study was performed on a representative sample of the Norwegian youth population (12-18 years; n=3237; response rate 45.2%). The proportion that never gambled was 17.6% and a majority (57.5%) gambled seldom, whereas 24.9% gambled weekly (36.2% of the males and 13.1% of the females). In relation to problematic gambling, the results showed that 1.76% had pathological gambling (2.79% in men and 0.69% in females) and 3.46% "at-risk" gambling. Problematic gambling (pathological gambling plus "at-risk" gambling) was 5.22% (7.82% of the males and 2.52% of the females). The group gambling frequently (at least weekly) was used to calculate pathological gambling and "at-risk" gambling. This resulted in high values, with 7.08% with pathological gambling (7.69% of males and 5.31% of females) and an additional 13.91% with "at-risk" gambling. The DSM-IV, with only 10 questions, gives a conservative estimate of pathological gambling. Slot machines proved the most popular game with 81.8%, followed by football tip (70.8%), Lotto (68.7%) and lotteries (39.4%). When it comes to problematic and pathological gambling, Lotto ranked high compared to other plays that were used more frequently. PMID- 12888410 TI - On the Far Eastern pattern of mortality. AB - Since the early 1980s, it has been accepted widely that there is a Far Eastern pattern of mortality, a pattern characterized by excessively high death rates among older men relative to death rates among younger men and among women. It has been regarded as a unique regional mortality pattern, applying primarily to Far Eastern populations. A re-examination of the mortality data of some Far Eastern populations reveals that changes in both age patterns of and sex differentials in mortality have been widely observed. Further, mortality patterns similar to the so-called Far Eastern mortality model have been found in many other populations. PMID- 12888411 TI - The contribution of HIV to fertility decline in rural Zimbabwe, 1985-2000. AB - HIV-related sub-fertility has been reported for those populations in sub-Saharan Africa in which contraceptive use is low. We use data from a retrospective survey in rural Zimbabwe and multivariate logistic regression models to show that recent birth rates and current pregnancy rates are also lower among HIV-positive women than among HIV-negative women in those African populations where contraceptive use is high. The fertility reduction is smaller than where contraceptive use is low because age at first sexual intercourse is later and birth rates at older ages are already low. Nevertheless, total fertility is approximately 8.5 per cent lower and HIV-associated sub-fertility may account for as much as one-quarter of fertility decline in Zimbabwe since the late 1980s. Mechanisms for HIV-associated sub-fertility in rural Zimbabwe include more frequent widowhood and divorce, reduced coital frequency, increased amenorrhoea, and possibly, pelvic inflammatory disease. Miscarriage appears to be a less important factor than elsewhere possibly because syphilis is rare. PMID- 12888414 TI - Pretransitional population control and equilibrium. AB - A persistent theme in much anthropological writing is the concept of the deliberate control of population numbers by hunter-gatherers as a means of achieving moderate family size, adequate nutrition, and constrained adult mortality. An analysis of the mix of theory and field evidence that led to this conclusion finds the case not proven. On the contrary, Malthusian constraints can operate, and probably did operate, to produce a hunter-gatherer society where most adults were reasonably robust and healthy even though child mortality was high and life expectancy short. The absence of population limitation in pre Neolithic times implies high mortality as well as high fertility, and weakens the argument positing a Neolithic mortality crisis. PMID- 12888415 TI - Annual mortality rates and excess deaths of children under five in Iraq, 1991-98. AB - Data from two parallel household surveys conducted in Iraq by UNICEF in 1999 show that under-5 mortality declined steadily from 1974 to 1990, reaching about 63 per 1,000 live births in the period 1986-90. It then rose dramatically to 118 per 1,000 in 1991, the year of the Gulf War. The number of 'excess' under-5 deaths (i.e., the number in excess of the number predicted from past trends) in Iraq between 1991 and 1998 was calculated assuming that, instead of the rates measured by the 1999 survey for this period, either (a) average mortality rates for the period 1986-90 had been maintained, or (b) mortality had continued to decline at the rate observed between 1974 and 1990. According to these calculations, the estimated number of excess deaths resulting from the Gulf War and its aftermath up to 1998 was between 400,000 (assumption a) and 500,000 (assumption b). PMID- 12888418 TI - Differential contributions of right and left brains to paw skill in right- and left-pawed female rats. AB - Paw preference and paw frequency was studied in female rats. Paw preference was assessed using a modified version of food reaching task in quadrupedal position. Of 68 rats, 56 (82.4%) were right-handed, 7 (10.3%) were left-handed, and 5 (7.4%) were mixed-handed. There were significantly more right-handers than left handers. The distribution of right minus left (L) paw reach (R + L = 50) was not U-shaped, it was J-shaped like in humans. Estrus cycle was a significant factor influencing the right-paw entry scores: most of left-handers were in estrus and proestrus, most of left -handers were in estrus, and most of mixed-handers were in postestrus. In right- and non-right- handers, the frequency of right-paw usage (right-hand skill) increased linearly with testing days, but the frequency of left-hand usage (left-hand skill) did not show significant changes with the successive testing days. Controlling for body weight and estrus only accentuated these results. It was concluded that distribution of hand preference in rats is J shaped and there is a right-sided population bias in handedness in rats like in humans. The results suggested that motor learning in paw skill is mainly involved the left brain in right- and mixed-handed rats, not the right brain: only the left brain has the inbuilt capacity for motor learning in female rats. Such an asymmetric cognitive control in an animal model may have a major impact in many aspects of biology in respect to normal functioning, superior talents, and disease (see Geschwind, 1985). PMID- 12888419 TI - Cerebrospinal fluid nitrite and malondialdehyde levels in patients with motor neuron disease. AB - Nitric oxide (NO) mediated oxidative damage may be involved in the pathogenesis of neuronal degeneration in motor neuron disease (MND). The present study was undertaken to evaluate the role of NO and oxidative stress in MND by estimating nitrite and malondialdehyde (MDA) levels in the cerebrospinal fluid (CSF) in 22 patients of MND and 20 control subjects suffering from neurological disorders not known to affect NO metabolism. There was no significant change in the CSF nitrite and MDA levels in MND. The nitrite and MDA levels did not have any significant correlation with age, duration of illness, or severity of disease. Univariate analysis of the clinical features in patients with MND and the nitrite levels revealed that two patients with a positive family history had significantly higher CSF nitrite levels as compared to those with a negative family history. There was no correlation between the CSF nitrite and MDA levels. Results of the present study did not indicate significant alterations in the MDA and NO levels in the CSF of MND patients. However, involvement of NO in MND with positive family history is suggested by the results obtained. PMID- 12888420 TI - Effect of immobilization and cold stress on visual evoked potentials. AB - The main aim of our research was to study the effects of immobilization and/or cold stress on amplitudes and latencies of visual evoked potentials (VEPs) and thiobarbituric acid reactive substances (TBARS). Forty healthy male albino rats, aged three months, were used. The rats were equally divided into four groups: Control group (C), the group exposed to cold stress (CS), the group exposed to immobilization stress (IS), and the group exposed to both cold and immobilization stress (CIS). Plasma corticosterone concentrations were significantly increased in all stress groups. Lipid peroxidation was increased in brain and retina of all stress groups as indicated by the significant increase in TBARS levels compared to the C group. Glutathione peroxidase (GSH-Px) activity in brain and retina increased in the CS group, but decreased in the IS group relative to the C group. GSH-Px activity also increased in the brain, but not in the retina in the CIS group with respect to the C group. The mean latencies of P1, N1, P2, N2, and P3 components were significantly prolonged in all stress groups compared with the C group. The results suggest that stress induced lipid peroxidation may influence VEPs. PMID- 12888421 TI - Stimulated microglia supernatants induced overexpression of neuronal nitric oxide gene in astrocytes. AB - The glia-glia, glia-neuron interactions are very complex and have not yet been clearly understood. Microglia gain reactivity in almost every type of brain insult and by affecting astrocytes and neurons, they determine the fate of damaged brain. In the present study we aimed to see the effect of stimulated microglia on nNOS and iNOS expression of astrocytes. The microglia cultures were stimulated with zymosanA and astrocytes were treated with the medium of microglia for 48 h. The results revealed that the astrocytes treated with microglia medium expressed more nNOS and iNOS than the ones treated with normal medium. PMID- 12888422 TI - Cell-mediated immune hypersensitivity is stronger on noninvolved side than involved side in patients with herpes zoster. AB - The difference between involved and noninvolved sides of the body in cell mediated immunity was investigated in this work; tuberculin reactions in two forearms were assessed in men and women with herpes zoster. The tuberculin reaction was smaller on the involved side than in the noninvolved side of the body in total sample, men and women. The results suggested that the lower activity of cell-mediated immune system in one side of the body may be related to the unilateral involvement of the zona-zoster infection. PMID- 12888423 TI - Effect of left- or right-handedness on nursing practice. AB - The aim of the present study was to evaluate the association of hand preference of nursing and the success of the nursing practice. The study included 24 students, all of whom attend different classes in the School of Nursing. The status of left- or right-handedness of the students included in the research was determined using Oldfield (Edinburgh Handedness Inventory) Index. The students were divided into two equal groups according to their right or left hand use. Then the students were categorized in four groups (Very good: 81-100, Average: 61 80, Pass: 50-60, and Fail: <50) according to the scores they received from the practice and theory tests of the lesson "Fundamentals of Nursing" the lesson requiring hand skills most. The test for the difference between two population proportions was used for statistical analysis. Left-handed nurses were found to be more successful than right-handed nurses in nursing practice. The difference between them was found significant (p < .01). It was concluded that left-handed nurses should never be forced to convert into right-handedness; instead they should be supported both verbally and in terms of equipment, and they should be motivated, because the data obtained proved that left-handedness means success. PMID- 12888424 TI - Mathematics and the gap junctions: in-phase synchronization of identical neurons. AB - A close consideration of some mathematical results with regards to neuronal synchronization mechanisms are examined. It is well known that intercellular coupling via gap junctions normally occurs between identical neurons, such as the coupling between inhibitory interneurons belonging to the same class. It is unknown why this should happen. The theory of coupled oscillators offers some explanations to answer the questions of the functional role of electrical interactions mediated by gap junctions and the necessity to couple identical neurons. The inference presented here from the mathematical results is that only if the cells are identical will their firing synchronize in-phase. Thus, we propose the concept that the functional role of gap junctional electrical coupling is to synchronize neurons in-phase and therefore this type of coupling will be found between neurons belonging to the same class. PMID- 12888425 TI - Hypothalamic digoxin and hemispheric chemical dominance: relation to alcoholic addiction, alcoholic cirrhosis, and acquired hepatocerebral degeneration. AB - The isoprenoid pathway produces three key metabolites--endogenous digoxin (modulate tryptophan/tyrosine transport), dolichol (important in N -glycosylation of proteins), and ubiquinone (free radical scavenger). It was considered pertinent to assess the pathway in alcoholic addiction, alcoholic cirrhosis, and acquired hepatocerebral degeneration. Since endogenous digoxin can regulate neurotransmitter transport, the pathway was also assessed in individuals with differing hemispheric dominance to find out the role of hemispheric dominance in its pathogenesis. In the patient group there was elevated digoxin synthesis, increased dolichol and glycoconjugate levels, and low ubiquinone and elevated free radical levels. There was also an increase in tryptophan catabolites and a reduction in tyrosine catabolites as reduced endogenous morphine synthesis from tyrosine. There was an increase in cholesterol:phospholipid ratio and a reduction in glycoconjugate level of RBC membrane in these groups of patients. The same patterns were obtained in individuals with right hemispheric chemical dominance. Alcoholic cirrhosis, alcoholic addiction, and acquired hepatocerebral degeneration are associated with an upregulated isoprenoid pathway and elevated digoxin secretion from the hypothalamus. This can contribute to NMDA excitotoxicity and altered connective tissue/lipid metabolism important in its pathogenesis. Endogenous morphine deficiency plays a role in alcoholic addiction. Alcoholic cirrhosis, addiction, and acquired hepato -cerebral degeneration occur in right hemispheric chemically dominant individuals. Ninety percent of the patients with alcoholic addiction, alcoholic cirrhosis, and acquired hepatocerebral degeneration were right-handed and left hemispheric dominant by the dichotic listening test. However, their biochemical patterns were similar to those obtained in right hemispheric chemical dominance. Hemispheric chemical dominance is a different entity and has no correlation with handedness or the dichotic listening test. PMID- 12888426 TI - Hypothalamic digoxin, hemispheric chemical dominance, and eating behavior. AB - The isoprenoid pathway produces an endogenous membrane Na+-K+ ATPase inhibitor, digoxin, which can regulate neurotransmitter and amino acid transport. Digoxin synthesis and neurotransmitter patterns were assessed in eating disorders. The patterns were compared in those with right hemispheric and left hemispheric dominance. The serum HMG CoA reductase activity, RBC membrane Na+-K+ ATPase activity, serum digoxin, magnesium, tryptophan catabolites (serotonin, quinolinic acid, strychnine, and nicotine), and tyrosine catabolites (morphine, dopamine, and noradrenaline) were measured in anorexia nervosa, bulimia nervosa, right hemispheric dominant, left hemispheric dominant, and bihemispheric dominant individuals. Digoxin synthesis was increased with upregulated tryptophan catabolism and downregulated tyrosine catabolism in those with anorexia nervosa and right hemispheric chemical dominance. Digoxin synthesis was reduced with downregulated tryptophan catabolism and upregulated tyrosine catabolism in those with bulimia nervosa and left hemispheric chemical dominance. The membrane Na+-K+ ATPase activity and serum magnesium were decreased in anorexia nervosa and right hemispheric chemical dominance while they were increased in bulimia nervosa and left hemispheric chemical dominance. Hypothalamic digoxin and hemispheric chemical dominance play a central role in the regulation of eating behavior. Anorexia nervosa represents the right hemispheric chemically dominant/hyperdigoxinemic state and bulimia nervosa the left hemispheric chemically dominant/hypodigoxinemic state. PMID- 12888427 TI - Hypothalamic digoxin and hemispheric chemical dominance in relation to the pathogenesis of bronchial asthma. AB - The isoprenoid pathway produces three key metabolites--digoxin (membrane sodium potassium ATPase inhibitor and regulator of neurotransmitter transport), dolichol (regulator of N-glycosylation of proteins), and ubiquinone (free radical scavenger). The isoprenoid pathway was assessed in patients with bronchial asthma. The pathway was also assessed in patients with right hemispheric, left hemispheric, and bihemispheric dominance to find out the role of hemispheric dominance in the pathogenesis of bronchial asthma. The pathway was upregulated with increase in digoxin synthesis in bronchial asthma. There was an increase in tryptophan catabolites and a reduction in tyrosine catabolites in patients with bronchial asthma. The ubiquinone levels were low and lipid peroxidation increased in these patients. There was increase in dolichol and glycoconjugate levels and reduction in lysosomal stability in these patients. The cholesterol:phospholipid ratio was increased and glycoconjugate levels were reduced in the membranes of these patients. The patterns noticed in bronchial asthma were similar to those in patients with right hemispheric chemical dominance. Bronchial asthma occurs in right hemispheric chemically dominant individuals. Ninety percent of the patients with bronchial asthma were right-handed and left hemispheric dominant by the dichotic listening test. But their biochemical patterns were similar to those obtained in right hemispheric chemical dominance. Hemispheric chemical dominance is a different entity and has no correlation with handedness or the dichotic listening test. PMID- 12888428 TI - Three dimensional modeling of the cerebrospinal fluid dynamics and brain interactions in the aqueduct of sylvius. AB - A computational fluid dynamics (CFD) method is presented to investigate the flow of cerebro-spinal fluid (CSF) in the cerebral aqueduct. In addition to former approaches exhibiting a rigid geometry, we propose a model which includes a deformable membrane as the wall of this flow channel. An anatomical shape of the aqueduct was computed from magnetic resonance images (MRI) and the resulting meshing was immersed in a marker-and-cell (MAC) staggered grid for to take into account fluid-structure interactions. The time derivatives were digitized using the Crank-Nicolson scheme. The equation of continuity was modified by introducing an artificial compressibility and digitized by a finite difference scheme. Calculations were validated with the simulation of laminar flow in a rigid tube. Then, comparisons were made between simulations of a rigid aqueduct and a deformable one. We found that the deformability of the walls has a strong influence on the pressure drop for a given flow. PMID- 12888429 TI - Can loaded interface characteristics influence strain distributions in muscle adjacent to bony prominences? AB - Pressure distributions at the interface between skin and supporting tissues are used in design of supporting surfaces like beds, wheel chairs, prostheses and in sales brochures to support commercial products. The reasoning behind this is, that equal pressure distributions in the absence of high pressure gradients is assumed to minimise the risk of developing pressure sores. Notwithstanding the difficulty in performing reproducible and accurate pressure measurements, the question arises if the interface pressure distribution is representative of the internal mechanical state of the soft tissues involved. The paper describes a study of the mechanical condition of a supported buttock contact, depending on cushion properties, relative properties of tissue layers and friction. Numerical, mechanical simulations of a buttock on a supporting cushion are described. The ischial tuberosity is modelled as a rigid body, whereas the overlying muscle, fat and skin layers are modelled as a non-linear Ogden material. Material parameters and thickness of the fat layer are varied. Coulomb friction between buttock and cushion is modelled with different values of the friction coefficient. Moreover, the thickness and properties of the cushion are varied. High shear strains are found in the muscle near the bony prominence and the fat layer near the symmetry line. The performed parameter variations lead to large differences in shear strain in the fat layer but relatively small variations in the skeletal muscle. Even with a soft cushion, leading to a high reduction of the interface pressure the deformation of the skeletal muscle near the bone is high enough to form a risk, which is a clear argument that interface pressures alone are not sufficient to evaluate supporting surfaces. PMID- 12888430 TI - The influence of the way the muscle force is modeled on the predicted results obtained by solving indeterminate problems for a fast elbow flexion. AB - A critical point in models of the human limbs when the aim is to investigate the motor control is the muscle model. More often the mechanical output of a muscle is considered as one musculotendon force that is a design variable in optimization tasks solved predominantly by static optimization. For dynamic conditions, the relationship between the developed force, the length and the contraction velocity of a muscle becomes important and rheological muscle models can be incorporated in the optimization tasks. Here the muscle activation can be a design variable as well. Recently a new muscle model was proposed. A muscle is considered as a mixture of motor units (MUs) with different peculiarities and the muscle force is calculated as a sum of the MUs twitches. The aim of the paper is to compare these three ways for presenting the muscle force. Fast elbow flexion is investigated using a planar model with five muscles. It is concluded that the rheological models are suitable for calculation of the current maximal muscle forces that can be used as weight factors in the objective functions. The model based on MUs has many advantages for precise investigations of motor control. Such muscle presentation can explain the muscle co-contraction and the role of the fast and the slow MUs. The relationship between the MUs activation and the mechanical output is more clear and closer to the reality. PMID- 12888431 TI - A finite element model for ice ball evolution in a multi-probe cryosurgery. AB - The ice formation in a water body is examined for the computation of temperature field, phase change and a moving ice-water interface whose location is not known a priori. This is classically referred to as the Stefan problem [Rubinstein, L.I. (1971) The Stefan Problem (American Mathematical Society, Providence, Rhode Island 02904]. Based on the Duvaut [Duvaut, G. (1973) "Resolution d'un probleme Stefan" C.R. Acad Sci. Paris 276, 1461-1463] transformation, the governing equations for heat conduction are formulated within a variational principle that is readily amenable to a standard finite element solution without remeshing. Numerical simulation results pertaining to the freezing of tumour tissue in a multi-cryoprobe cryosurgery are presented. These results lend both quantitative and graphical support to the current empirical standards of "effective therapy" in view of refining clinical applications. PMID- 12888432 TI - Finite element modeling of trabecular bone damage. AB - This paper presents a finite element-based, computational model for analysis of structural damage to trabecular bone tissues. A modulus reduction method was formulated from elasto-plasticity theory, and was used to account for site specific trabecular bone tissue damage. Trabecular bone tissue damage is illustrated using a large-scale, anatomically accurate, two-dimensional, microstructural finite element model of a human thoracic vertebral body. Four models with varying specifications for damage accumulation were subjected to compressive loading and unloading cycles. The numerical results and experimental validation demonstrated that the modulus reduction method reproduced the non linear mechanical behaviour of vertebal trabecular bone. The iterative computational approach presented provides a methodology to study trabecular bone damage, and should provide researchers with a computational approach to study bone fracture and repair and to predict vertebral fragility. PMID- 12888433 TI - A density distribution algorithm for bone incorporating local orthotropy, modal analysis and theories of cellular solids. AB - An algorithm for bone remodeling is presented which allows for both a redistribution of density and a continuous change of principal material directions for the orthotropic material properties of bone. It employs a modal analysis to add density for growth and a local effective strain based analysis to redistribute density. General re-distribution functions are presented. The model utilizes theories of cellular solids to relate density and strength. The code predicts the same general density distributions and local orthotropy as observed in reality. PMID- 12888434 TI - Mild cognitive impairment. PMID- 12888435 TI - The role of volumetric MRI in understanding mild cognitive impairment and similar classifications. AB - We review nineteen empirical studies of mild cognitive impairment (MCI), age associated memory impairment (AAMI) and related classifications reporting volumetric data on the hippocampus, entorhinal cortex and amygdala. Studies varied considerably in terms of the selection of participants, sample characteristics, the definitions of regions of interest and normalization techniques. Effect sizes for differences in left hippocampal volume and right hippocampal volumes of AAMI, MCI and pre-clinical dementia groups compared with controls ranged from 0.47 to 1.34. Effect sizes for left and right hippocampal volumes for Alzheimer's disease (AD) versus control were 1.88 and 1.75 respectively. Longitudinal results confirm that initial hippocampal volume is predictive of conversion to AD. Greater standardization in methodology and the development of normative age-referenced databases of regional brain volumes is required. PMID- 12888436 TI - The detection of mild cognitive impairment in the general practice setting. AB - Results from a general practice study of cognitive impairment are used to construct guidelines for the detection of mild cognitive impairment (MCI) in general practice. The role of the general practitioner in the diagnosis of MCI is discussed and the potential feasibility of general practice screening is evaluated with data from a two-year prospective study conducted in the Montpellier region of France. Neuropsychological tests with the highest predictive value for dementia conversion and suitable for use in general practice, were determined by logistic regression modelling. An examination comprising three tests (delayed auditory verbal recall, verbal fluency and visuospatial construction), were found to give a specificity of 99% and sensitivity of 73%. Reports from other research centres suggest that MCI detection should not be limited to cognitive performance alone. It is suggested that proxy observations of behavioural change and information relating to loss of ability to perform activities of daily living should also be used to improve sensitivity and also to provide information needed in patient management. It may be potentially feasible for the family practitioner to verify cognitive complaints and to screen for MCI with a high degree of accuracy using a brief test battery derived from empirical observations in population studies. PMID- 12888437 TI - Cognitive decline in high-functioning older adults: reserve or ascertainment bias? AB - The detection of mild cognitive impairment and dementia in high-functioning older adults can be difficult. It has also been observed that high-functioning persons show a lower prevalence of dementia than low-functioning persons. Three alternative explanations for this observation have been proposed in the literature: brain reserve capacity (BRC), cognitive reserve, and ascertainment bias. With data from a prospective, population-based study of incident dementia, the Canadian Study of Health and Aging (CSHA), we classified participants as being high- (HF) or low-functioning (LF) in three ways: educational and occupational attainment, and estimated premorbid IQ. We observed that fewer HF older adults were diagnosed with dementia after five years, which is in accordance with both the BRC and cognitive reserve models. Contrary to expectations, no difference on rate of memory deterioration was observed between those HF and LF persons who exhibited mild cognitive impairment at CSHA-1. However, HF persons who subsequently were diagnosed with dementia (CSHA-2) showed more rapid decline on five of the six memory measures over time than did LF persons diagnosed with dementia at CSHA-2. When performance on measures of memory functioning at CSHA-1 was examined for highly educated older adults, significantly more of those with dementia at CSHA-2 (n = 59) had scores falling within or below the average range in comparison to normative standards than those who continued to show no cognitive impairment (n = 145). Our findings suggest that the lower incidence of dementia for HF persons may be primarily the result of ascertainment bias, not underlying differences in brain or cognitive reserve. PMID- 12888438 TI - Mild cognitive impairment in the oldest old: a comparison of two approaches. AB - The main purpose of the current investigation was to examine the predictive utility of two sets of criteria for mild cognitive impairment (MCI). A second purpose was to determine the incidence of MCI in a longitudinal, population-based sample of the oldest old in Sweden. The participants were from the larger 'Origins of Variance in the Old-Old: Octogenarian Twins' (OCTO-Twin) study, identified through the Swedish Twin Registry. Participants were initially aged 80 or above with a mean age of 83 and were re-examined after two, three, six and eight years. The sample for this study consisted of 263 elders that were randomly selected singletons from twin pairs. Mild cognitive impairment was rated at baseline using both Petersen'sand Ritchie's criteria. Petersen's criteria emphasize memory and memory complaint whereas Ritchie's criteria use a broader set of cognitive indicators. The incidence rate was comparable to that of other studies, however, neither set of criteria predicted subsequent dementia. The failure to confirm subsequent dementia suggests that there may be many sources of MCI in very late life besides incipient dementia. The presence of a key informant, as well as the weighting of performance across domains by an experienced clinician, both of which are hard to quantify, may be the key elements that give predictive values to MCI in prior studies. PMID- 12888439 TI - Different attitudes when handling aggressive behaviour in dementia--narratives from two caregiver groups. AB - This study highlights the experiences of 15 formal caregivers, during interactions with elderly residents suffering from dementia disease and showing aggressive behaviour. The purpose was to study caregivers' reflections about and attitudes to behavioural and psychiatric symptoms of dementia (BPSD) and how they dealt with the symptoms. This was done by comparing care units with high or low levels of aggressive behaviour in residents. A phenomenological-hermeneutic approach was used for the analysis of the interviews. The main themes that emerged were: a need for balance between demands and competence; and a need for support. The findings indicated the importance of a balance for the residents as well as for the caregivers, if a positive relationship was to develop. Furthermore, caregivers stated that support was crucial, not only for the residents but also for themselves, if they were expected to cope with demanding situations. Different types of support were necessary and included: confirmation, feedback, and supervision. Residents who feel appreciated and respected may be less likely to act out their frustrations in an inappropriate manner. Caregivers who strive to understand the meaning behind a resident's behaviour and who master the necessary care-giving skills, and their implementation, could be more successful at curbing distressing behaviour, than caregivers who act merely in a custodial role. PMID- 12888440 TI - Addressing the informational needs of healthcare professionals in dementia care: an explorative study in a Chinese society. AB - Many healthcare professionals in Hong Kong are now working in dementia care, but little is known about their informational needs in this area. This study explored the level of knowledge about dementia that healthcare professionals perceived themselves as having, and the informational needs of dementia care. 320 respondents, who were primarily nurses, social workers, and rehabilitation therapists, completed a questionnaire designed for this study. While 65% reported having sufficient knowledge of dementia, only 40% expressed having adequate knowledge of dementia-related services. Respondents who had relevant clinical experience demonstrated a significantly higher self-perceived knowledge level of dementia (p=0.000) and dementia-related services (p=0.000). The respondents ranked information leaflets (68%), the mass media (65%), and workshops (53%) as the three popular sources for obtaining dementia-related information. Similar to their western counterparts, over 75% of respondents expressed the need to lean more about clinical management and service provisions for people with dementia and their families. To enhance the knowledge of healthcare professionals about dementia care, it is recommended that healthcare professionals be encouraged to participate in continuing education programmes, and that education/resource centres for dementia care be developed to address the learning needs of local practitioners. PMID- 12888441 TI - Relationships between altruistic activity and positive and negative affect among low-income older adult service providers. AB - Self-reported altruistic activity was examined as a predictor of positive and negative affective states among a sample of 115 low-income older adults who were actively providing services to others within a federally subsidized service delivery program (Foster Grandparents and Senior Companions). Results indicated that altruistic activity was predictive of positive, but not negative affect. It was found that altruistic activity was a significant predictor of positive affect after controlling for relevant demographic variables including social support and income. The theoretical and clinical implications of this research are discussed. PMID- 12888442 TI - Sharing a diagnosis of dementia--learning from the patient perspective. AB - Sharing a diagnosis of dementia is a key focus of debate and research in the field of dementia care. The pivotal role of the GP in the early identification and subsequent management of dementia has long been recognized and whilst diagnostic practice is improving, the knowledge gained from research exploring the patient perspective is insufficiently absorbed. This paper presents the evidence from the two perspectives of the diagnostic relationship: the views of GPs around giving a diagnosis and the views of patients on receiving one. The authors draw upon their own projects-as well as wider research-in suggesting ways that diagnostic practice can be improved by taking account of the patient perspective. The learning can be distilled around the dimensions of the process, approach and nature of disclosure with evidence particularly challenging the 'myths' held by many GPs around the value of early diagnosis. Whilst much work still needs to be done to incorporate the patient perspective in dementia care, there is clear evidence that patient oriented research has much to offer the development of disclosure practice in primary care to the benefit of patients, their families and GPs. PMID- 12888443 TI - Correlates of everyday competence in Chinese older adults. AB - Whether older adults can maintain levels of adaptation that allow continuation of independent living is necessarily contingent upon the maintenance of levels of everyday competence. This study identifies factors correlated to everyday competence measured by a Chinese version of the Direct Assessment of Functional Abilities among Hong Kong Chinese elderly people. The respondents were 393 people aged 60 years or older from a cross-sectional study of a representative community sample of the elderly population in Hong Kong and they were interviewed in a face to-face format. In multiple regression analyses, we found that self-rated health, sight, and global cognitive ability were positively associated with everyday competence whereas the presence of arthritis had a negative impact on the everyday competence. All these findings are consistent with previous Western studies. PMID- 12888444 TI - Molecular markers of endocrine disruption in aquatic organisms. AB - A wide range of organic contaminant compounds prevalent in the aquatic environment has been shown to exhibit hormone-disrupting activity. The actual potency of such compounds are low compared with endogenous hormones, such as 17beta-estradiol, but may still produce detrimental biological effects. Induced hormone levels are routinely measured using commercial testing kits, though these fail to relate to actual effects. Field and laboratory studies on the biological effects of environmental estrogens have, in the past, largely relied on assays of vitellogenin (vtg) induction in male fish, reduced growth in testes formation, and intersex incidence. Here, we critically review the current and potential application of molecular techniques in assessing the adverse biological reproductive effects of endocrine-disrupting chemicals in aquatic organisms. The role of fish (estrogen, androgen, and progestogen) hormone receptors and invertebrate (ecdysone) hormone receptor, egg production (vtg and chorion) proteins, steroid biosynthesis enzymes (aromatase, sulfotransferase, and hydroxysteroid dehydrogenase), DNA damage, apoptosis, and their potential development as biomarkers are discussed in turn. In each case, the sequences characterized are presented and homologies across species are highlighted. Molecular methods of gauging vtg and zona radiata (ZR) expression and protein concentrations have included immunoassay and reverse transcription polymerase chain reaction (RT-PCR). Suggestions for the isolation for key gene expression products (aromatase, ZR, and vtg, for instance), from a wider range of fish species using degenerate primers, are given. Endocrine disruption in invertebrates has received less attention compared with fish, partly because the knowledge regarding invertebrate endocrinology is limited. Here we review and suggest alternate isolation strategies for key players in the imposex induction process: vitellin (Vn), aromatase, and Ala-Pro-Gly-Trp (APGW) amide neurohormone. Current molecular-level techniques rely on ligand-binding assays, enzyme-linked immunosorbent assay (ELISA), and, more recently, gene expression. In the future, more reliance will be placed on the development of gene expression assays using reporter systems combined with cross-species PCR-based or polyclonal antibody based assays. We discuss the use of recombinant receptors as a means of primary screening of environmental samples for estrogenicity and antiestrogenicity, which avoids species and seasonal variation in receptor response to ligand binding, a recognized problem of earlier bioassays. Most exciting is the potential that microarray and proteomics approaches have to offer. Such techniques are now used routinely in medical research to identify specific genes and proteins affected by treatment with endocrine disruptors, including estradiol. The technique has yet to be used to screen aquatic organisms, but it has the potential to implicate previously unsuspected estradiol-sensitive genes that may later become molecular markers of endocrine disruption. PMID- 12888445 TI - Developing risk-based target concentrations for carcinogenic polycyclic aromatic hydrocarbon compounds assuming human consumption of aquatic biota. AB - As part of the remediation process at a former creosote-handling facility in Washington, target groundwater concentrations were developed as goals for the planned cleanup efforts. Considering state regulatory requirements and site specific conditions, these concentrations were established to protect surface water in the lake adjacent to the site. These risk-based values were calculated assuming that chemicals will (1) be transported in groundwater, (2) discharge into the lake, and (3) be taken up by aquatic organisms that may be consumed by humans. Among the primary chemicals driving remediation decisions at this site are carcinogenic polycyclic aromatic hydrocarbon (cPAH) compounds, which have limited environmental mobility and are metabolized by many types of potentially edible aquatic organisms. This work included assessing the validity for cPAH compounds of the required default regulatory assumptions and deriving alternative risk-based concentrations. These analyses focused on factors that would modify the generic assumption regarding bioconcentration of cPAH compounds in aquatic biota and influence bioavailability of cPAH compounds to humans consuming the biota. Modifications based on these factors and the use of toxicity equivalency factors resulted in alternative risk-based concentrations for individual cPAH compounds that ranged from approximately 7 to 700 times greater than the default value of 0.03 microg/l. PMID- 12888446 TI - Nasal epithelial and inflammatory response to ozone exposure: a review of laboratory-based studies published since 1985. AB - This article summarizes biological events in human and animal nasal epithelium after short- and long-term exposure to ozone, the principal agent in photochemical smog. Despite anatomical and histological interspecies differences, ozone exposures resulted in common nasal qualitative alterations with an anterior posterior gradient of phenomena occurring immediately, and with a lag time postexposure: epithelial disruption and increased permeability, inflammatory cell influx, and proliferative and secretory responses. Described mechanisms of toxicity included a direct effect of ozone on epithelial lining fluid and cellular membranes and the subsequent release of cytokines and cyclooxygenase and lipoxygenase products. An indirect effect of ozone was indicated by a decreased mucociliary clearance, free radicals production interacting with a gene promoting factor, and increased DNA synthesis. Studies highlighted the pivotal role of activated neutrophils and mast cells leading to the release of deleterious enzymes (tryptase, eosinophil cationic protein) and numerous cytokines. Experiments performed with ozone exposure/allergen challenge reported that, besides the intrinsic deleterious properties of ozone, it also had a priming effect on the late-phase response to allergen challenge, providing new insights into the pathophysiology of respiratory allergic diseases. PMID- 12888447 TI - Tracheal stenosis depends on the extent of cartilaginous injury in experimental canine model. AB - To test the hypothesis that the development of tracheal stenosis would depend on the extent of tracheal cartilaginous injury, either 90 degrees (n=6) or 180 degrees (n=6) of anterior wall of 4 tracheal rings were cauterized in 12 mongrel dogs using Nd-YAG laser. Beginning at 3 weeks after cauterization, 180 degrees tracheal injury resulted in life-threatening tracheal stenosis whereas 90 degrees injury did not. Gross and microscopic examinations showed that after 180 degrees injury, tracheal stenosis was accompanied by the loss of tracheal cartilage and dense fibrosis. These results indicate that tracheal stenosis depends on the extent of tracheal injury in experimental canine model. PMID- 12888448 TI - Contractility and phenotype of human bronchiole smooth muscle after prolonged fetal bovine serum exposure. AB - Culture of dispersed airway smooth muscle (ASM) cells with fetal bovine serum (FBS) induces rapid growth and modulation from a contractile to a synthetic phenotype, but this may be an artificial situation due to loss of tissue architecture. In this study, structural, functional and biochemical changes of ASM were examined in human intact bronchiole ring segments (200 to 600 micro m internal diameter) after organ culture for up to 6 days in 10% FBS or in D-STIM, an FBS-free medium formulated to maintain a contractile phenotype. ASM content was unchanged after culture for 3 or 6 days with D-STIM or FBS compared with fresh tissues. However, by 6 days culture with FBS reduced the maximum developed contraction to several agonists (carbachol, histamine, and KCl). Smooth muscle (sm)-alpha-actin and sm-myosin heavy chain (MHC) isoform 1 expression was similar for all culture conditions, though FBS reduced calponin and metavinculin content. Nonmuscle (nm) proteins, including total vinculin, beta-actin, and nm-MHC, were unchanged. Thus, ASM in human intact bronchioles maintains its functional, biochemical, and morphometric properties after culture in FBS-free (D-STIM) medium for at least 6 days, and for 3 days when cultured with FBS. These findings in organ culture may reflect more closely the in vivo situation in which tissue architecture is better preserved over cell culture, and may provide a basis for examining long-term effects of trophic or contractile stimuli on intact ASM in vitro that contribute to airway hyperresponsiveness and remodeling. PMID- 12888449 TI - Transforming growth factor beta contributes to lung leak in rats given interleukin-1 intratracheally. AB - Interleukin-1 (IL-1) is increased in lung lavages obtained from patients with acute lung injury (ALI) and administering recombinant human IL-1alpha (rhIL 1alpha) (50 ng) intratracheally causes an acute, neutrophil-dependent, oxidative lung leak in rats that closely resembles human ALI. In the present work, the authors tested the hypothesis that transforming growth factor beta (TGFbeta) contributes to the lung inflammation and injury that develops in rats given IL-1 intratracheally. They found that intravenous administration of a monoclonal antibody to TGFbeta (1.D.11.16, 0.5 mg/kg) attenuated lung injury responses, specifically lung leak index, lung lavage protein concentrations, and blood oxygenation abnormalities, that are observed 5 hours after intratracheal instillation of IL-1 in rats, but did not decrease indices of lung inflammation, specifically myeloperoxidase (MPO) activity in lung tissue, neutrophil counts in lung lavage, and cytokine-induced neutrophil chemoattractant (CINC) levels in lung lavage, in rats given IL-1 intratracheally. The results suggest that TGFbeta contributes to lung leak, but not lung inflammation, following intratracheal administration of IL-1 in rats. PMID- 12888450 TI - Differential production of metalloproteinases after instilling various urban air particle samples to rat lung. AB - Lung injury and inflammation are associated with exposure to various types of particulate air pollutants. The present study was used to determine whether metalloproteinases (MMPs) are secreted after instilling dust samples into the lung, and to relate levels of specific MMPs to different fractions of the ambient air particle sample EHC-93. Rats received an intratracheal injection of 5 mg dust samples in 0.5 ml water and were killed at intervals from 4 hours to 28 days later particle samples were EHC-93 whole dust, and the insoluble, leached, and soluble fractions of the same dust. Samples prepared from EHC-2K dust were also used, as were solutions of zinc and copper chloride. All samples induced inflammation as measured by increased inflammatory cells in bronchoalveolar lavage (BAL) fluid; the highest levels were found 1 to 3 days after instilling the whole dust. This dust also induced production of MMP-2 and MMP-9 as shown in zymograms. The leached dust induced predominantly MMP-9, which was maximal at 4 hours and 1 day. In contrast, the soluble fraction induced almost exclusive 4 MMP 2, also maximal at 4 hours and 1 day; this enzyme was also produced in response to soluble zinc, the most prevalent soluble metal in the EHC samples. The results demonstrate the rapid production and secretion of MMPs in the lung after particle deposition. A differential pattern of MMP production is seen with MMP-9, likely from inflammatory cells, being produced in response to the insoluble particles, and MMP-2, likey from epithelial cells, being produced in response to the water soluble fraction of the atmospheric dust. PMID- 12888451 TI - Acid exposure potentiates intercellular adhesion molecule-1 and e-cadherin expression on A549 alveolar lining epithelial cells. AB - Recruitment of neutrophils into the alveoli plays a major role in the pathogenesis of acid-induced pneumonitis. Preliminary data suggest that alteration in the expression of cellular adhesion molecules on the airway epithelial cells may play an important role in the recruitment of neutrophils following acid-induced lung injury. The aim of this study was to evaluate the change in the surface expression of intercellular adhesion molecule-1 (ICAM-1), E cadherin, and vascular cell adhesion molecule -1 (VCAM-1) on acid-exposed A549 alveolar lining epithelial cells by flow cytometry and confocal laser microscopy. Acid exposure changed cell morphology, increased cell adhesion after trypsin-EDTA treatment, and up-regulated the expression of ICAM-1 and E-cadherin but not of VCAM-1. The up-regulation of ICAM-1 expression will induce the dysfunction of epithelial cells with or without accumulation of neutrophils in air spaces. Because the distribution of E-cadherin in acid-exposed A549 cells was at the sites where the cells attached to culture dish but not at the intercellular junctions between adjoining cells, up-regulated expression of E-cadherin will rather result in alterations of epithelial morphology and function of epithelial barrier. In addition, pentoxifylline suppressed the up-regulation of ICAM-1 and E cadherin expression and may therefore attenuated the airway inflammation in acid induced pneumonitis. PMID- 12888453 TI - Leukocyte trafficking to the lung. Abstracts of the British Association for Lung Research summer meeting. Dublin, Ireland, 28-30 August 2002. PMID- 12888452 TI - Regulation of toll-like receptor 2 and 4 gene expression in murine alveolar macrophages. AB - The authors investigated the regulation of toll-like receptor (TLR) TLR2 and TLR4 gene expression in alveolar macrophage (AM) in response to lipopolysaccharide (LPS) or proinflammatory cytokines in vitro. Treatment of a murine AM cell line, MH-S, with LPS, tumor necrosis factor (TNF)-alpha and interleukin (IL)-1beta significantly increased TLR2 mRNA expression, whereas TLR4 mRNA expression remained constant. LPS-mediated TLR2 mRNA up-regulation was attenuated by inhibition of p38 kinase (with SB203580) or nuclear factor (NF)-kappaB (with sulfasalazine or SN-50), but not by inhibition of extracellular signal-regulated kinase (with PD98059) or c-Jun N-terminal kinase (with SP600125), suggesting that LPS may induce TLR2 mRNA expression through p38 kinase and NF-kappaB activation. These results indicate that TLR2 expression up-regulated in AM in response to bacterial respiratory infections may render AM responsive to TLR2 ligands, which may accelerate the innate immunity against pathogens in the lung. PMID- 12888455 TI - What is female sexual dysfunction? PMID- 12888457 TI - Travel and health. Reducing your risk for altitude sickness. PMID- 12888456 TI - I can see clearly now...sort of. Will eye exercises eliminate your need for corrective lenses? No, and here's why. PMID- 12888458 TI - Physical exercise sharpens the brain. PMID- 12888459 TI - By the way, doctor. You've written about the dangers of getting too much vitamin A from retinol. Do you know of any multivitamin brands that tell you how much retinol you're getting? PMID- 12888460 TI - By the way, doctor. I don't hear nearly as much about hair loss in women as in men. But I'm 65 and have watched my hair thin considerably over the past five or six years. Is there anything I can do about it? What about Rogaine? PMID- 12888461 TI - Salt and your health: the role of sodium. The first of two parts. PMID- 12888462 TI - The prostate biopsy. PMID- 12888463 TI - Alcohol and your waistline. PMID- 12888464 TI - On call. Breast cancer seems to run in my wife's family. She is healthy, I'm glad to say, but her mother died of it, and her sister needed surgery and chemo. Worst of all, our 44-year-old daughter just had a mastectomy and is waiting to hear if she needs more treatment. We also have two sons, ages 46 and 40. My question is, do they have anything to worry about? PMID- 12888465 TI - Cancer and the mind. Cancer does not come from thoughts or feelings, but psychosocial treatment can help cancer patients. PMID- 12888466 TI - Prisoners of mental illness. PMID- 12888467 TI - Fit for thinking. PMID- 12888468 TI - There is a new antidepressant escitalopram (Lexapro) that is a cousin of citalopram (Celexa). How are these two different and does the newer drug have any advantages? PMID- 12888469 TI - DEET is hard to beat. The chemical discovered over a half-century ago is still the best mosquito repellent--and safer than you might think. PMID- 12888470 TI - Herbal medicines: Newer rules, familiar problems. Proposed regulations may improve quality but still won't require proof of safety and effectiveness. PMID- 12888471 TI - The ultimate antiperspirant. Botox injections have many uses, including the treatment of excessive sweating. PMID- 12888472 TI - Blood pressure normal? Maybe now it isn't. Prehypertension has replaced normal and high-normal in new government guidelines for high blood pressure. PMID- 12888473 TI - By the way, doctor. My new cardiologist upped my Zocor from 10 mg to 40 mg daily. I've had bypass and valve surgery but was feeling fine, and as far as I know, my cholesterol level is okay. Does it make sense to increase the dose that much? PMID- 12888474 TI - By the way, doctor. I have several medications that are well beyond their expiration dates. Does that mean that they won't work--or are dangerous? PMID- 12888475 TI - Dangerous duo--heart disease and kidney disease. PMID- 12888476 TI - Time to wave goodbye to morning surge in blood pressure. A big spike in blood pressure after getting out of bed should trigger a reevaluation of your medications. PMID- 12888477 TI - At peace with warfarin. 8 tips for staying on the safe side of this important "blood thinner". PMID- 12888478 TI - Test opens window into the failing heart. A hormone made by an overworked or damaged heart offers important clues to heart function and new avenues for treatment. PMID- 12888479 TI - Artery picture not worth 1,000 words. PMID- 12888480 TI - Exercise intensity: feeling what's right. PMID- 12888481 TI - PREMIER results. PMID- 12888482 TI - Ask the doctor. I've been seeing ads for a test called IMT that is supposed to spot heart disease and tell how serious it is, even if you don't have any symptoms. Is this for real? PMID- 12888483 TI - Ask the doctor. I am an 80-year-old woman who had a triple bypass many years ago. The veins of my legs have closed, and my feet and legs are swollen and painful. My doctor explained that the blood flows down the legs but does not flow back up. She also said that nothing can be done for it. Is this true? PMID- 12888484 TI - Pheromone-induced anorexia in male Syrian hamsters. AB - Transition from long days (LDs) to short days (SDs) triggers seasonal obesity in Syrian hamsters. We report here that SD-exposed males housed near females exhibit obesity resistance, episodic weight loss, and reduced adiposity. Negative energy balance is achieved by reduced eating, elevated motor activity, and increased caloric efficiency without metabolic compensation. Circulating leptin, insulin, testosterone, corticosterone, and cortisol are normal or reduced in obesity resistant hamsters. When males are housed in chambers that block physical, visual, and auditory, but not pheromonal, signals from females, resistance to seasonal obesity persists. Moreover, inhalation of extracts from pheromone releasing flank glands of females suppresses eating and weight gain in SD-exposed males. This novel phenomenon, pheromone-induced anorexia, shows that female pheromones play a critical role in the seasonal energy balance of male hamsters. These findings provide a model to study neural and endocrine mechanisms that underlie eating disorders. PMID- 12888485 TI - BMPs and BMPRs in chicken ovary and effects of BMP-4 and -7 on granulosa cell proliferation and progesterone production in vitro. AB - Bone morphogenetic proteins (BMPs) and their receptors (BMPRs) are now known to have important roles in mammalian ovarian folliculogenesis. This study determined the expression of the mRNA encoding for BMPs and their receptors in the chicken ovary and explored possible roles for them. The expression of the mRNA for BMP-2, -4, -6, -7, and BMPR-IA, -IB, and -II was determined and quantified by a semiquantitative RT-PCR. The mRNAs for all the BMPs and receptors determined were present in both the granulosa (G) and theca (T) cells of the F1, F2, and F3 follicles. All BMP mRNAs increased in G cells with follicular development, whereas only BMP-7 mRNA had this trend in the T cells. BMP-2, -4, and -6 mRNAs in T were similar between follicles. BMPR-IA mRNA was similar in F2G and F3G but lower in F1G. BMPR-IB mRNA was similar in G of all follicles, and BMPR-II mRNA increased with development. In the T, each receptor subtype showed equal distribution between follicles. mRNA levels for BMPR-IB and -II were higher in G than in T, suggesting that the G is a major target for BMPs. BMP-4 and -7 stimulated basal, IGF-I-, and gonadotropin-stimulated progesterone production by cultured G cells, with differential responses between cells from the F1 and F3/4. This suggests involvement in follicular differentiation. BMP-4 and -7 reversed the inhibitory effects of transforming growth factor (TGF)-alpha on basal and gonadotropin-stimulated G cell progesterone production, with greater effect in the F1 than in the F3/4. This effect suggests an important role for BMPs interacting with TGF-alpha in modulating the effects of gonadotropins and IGF-I on follicular differentiation. Finally, BMP-7 stimulated G cell proliferation, but BMP-4 inhibited TGF-alpha+ IGF-I- and/or FSH-stimulated G cell proliferation, suggesting a role in the control of follicular growth during development. These effects of BMP-4 and -7 on the G cell function showed relationships with the expression levels of the BMPs and the BMPR-II. PMID- 12888486 TI - A statistical model of diurnal variation in human growth hormone. AB - The diurnal pattern of growth hormone (GH) serum levels depends on the frequency and amplitude of GH secretory events, the kinetics of GH infusion into and clearance from the circulation, and the feedback of GH on its secretion. We present a two-dimensional linear differential equation model based on these physiological principles to describe GH diurnal patterns. The model characterizes the onset times of the secretory events, the secretory event amplitudes, as well as the infusion, clearance, and feedback half-lives of GH. We illustrate the model by using maximum likelihood methods to fit it to GH measurements collected in 12 normal, healthy women during 8 h of scheduled sleep and a 16-h circadian constant-routine protocol. We assess the importance of the model components by using parameter standard error estimates and Akaike's Information Criterion. During sleep, both the median infusion and clearance half-life estimates were 13.8 min, and the median number of secretory events was 2. During the constant routine, the median infusion half-life estimate was 12.6 min, the median clearance half-life estimate was 11.7 min, and the median number of secretory events was 5. The infusion and clearance half-life estimates and the number of secretory events are consistent with current published reports. Our model gave an excellent fit to each GH data series. Our analysis paradigm suggests an approach to decomposing GH diurnal patterns that can be used to characterize the physiological properties of this hormone under normal and pathological conditions. PMID- 12888487 TI - Mechanisms of P/CAF auto-acetylation. AB - P/CAF is a histone acetyltransferase enzyme which was originally identified as a CBP/p300-binding protein. In this manuscript we report that human P/CAF is acetylated in vivo. We find that P/CAF is acetylated by itself and by p300 but not by CBP. P/CAF acetylation can be an intra- or intermolecular event. The intermolecular acetylation requires the N-terminal domain of P/CAF. The intramolecular acetylation targets five lysines (416-442) at the P/CAF C terminus, which are in the nuclear localisation signal (NLS). Finally, we show that acetylation of P/CAF leads to an increment of its histone acetyltransferase (HAT) activity. These findings identify a new post-translation modification on P/CAF which may regulate its function. PMID- 12888488 TI - Functional characterization of cis- and trans-regulatory elements involved in expression of phospholipid hydroperoxide glutathione peroxidase. AB - Phospholipid hydroperoxide glutathione peroxidase (phGPx) is a member of the seleno glutathione peroxidase family that is comprised of five selenoproteins capable of reducing hydroperoxy lipids to the corresponding alcohols. The enzyme has been implicated in antioxidative defense, but its high expression level in testicular tissue suggests a more specific function during sperm maturation. The phGPx is encoded for by a joint sperm nucleus/phGPx gene (sn/phGPx) and can be expressed as a mitochondrial or cytosolic isoform. Although sn/phGPx genes have been cloned from various mammalian species expression regulation of the enzyme has not been studied in detail. We investigated the 5'-flanking region of the murine sn/phGPx gene and observed basic promoter activity in a 200 bp region localized immediately upstream of the translational initiation site of the cytosolic isoform (3'-ATG). DNase protection assays indicated the presence of five distinct protein-binding regions and electrophoretic mobility shift assays and supershift experiments revealed binding of stimulating protein 1 (SP1), nuclear factor Y (NF-Y) and members of the SMAD family. Site-directed mutagenesis of the consensus binding sequences abolished in vitro transcription factor binding. Expression of reporter genes was most effectively impaired when SP1/SP3 and NF-Y binding site-deficient constructs were tested. Chromatin immunoprecipitation suggested the in vivo relevance of these transcription factors. Our data indicate that the basic phGPx promoter constitutes a 200 bp oligonucleotide, which is localized immediately upstream of the 3'-ATG and involves functional SP1/SP3, NF-Y and SMAD binding sites. The corresponding trans regulatory proteins may contribute to differential expression regulation of the mitochondrial and cytosolic phGPx isoforms. PMID- 12888489 TI - Functional conservation between members of an ancient duplicated transcription factor family, LSF/Grainyhead. AB - The LSF/Grainyhead transcription factor family is involved in many important biological processes, including cell cycle, cell growth and development. In order to investigate the evolutionary conservation of these biological roles, we have characterized two new family members in Caenorhabditis elegans and Xenopus laevis. The C.elegans member, Ce-GRH-1, groups with the Grainyhead subfamily, while the X.laevis member, Xl-LSF, groups with the LSF subfamily. Ce-GRH-1 binds DNA in a sequence-specific manner identical to that of Drosophila melanogaster Grainyhead. In addition, Ce-GRH-1 binds to sequences upstream of the C.elegans gene encoding aromatic L-amino-acid decarboxylase and genes involved in post embryonic development, mab-5 and dbl-1. All three C.elegans genes are homologs of D.melanogaster Grainyhead-regulated genes. RNA-mediated interference of Ce-grh-1 results in embryonic lethality in worms, accompanied by soft, defective cuticles. These phenotypes are strikingly similar to those observed previously in D.melanogaster grainyhead mutants, suggesting conservation of the developmental role of these family members over the course of evolution. Our phylogenetic analysis of the expanded LSF/GRH family (including other previously unrecognized proteins/ESTs) suggests that the structural and functional dichotomy of this family dates back more than 700 million years, i.e. to the time when the first multicellular organisms are thought to have arisen. PMID- 12888490 TI - Divalent metal-dependent catalysis and cleavage specificity of CSP41, a chloroplast endoribonuclease belonging to the short chain dehydrogenase/reductase superfamily. AB - CSP41 is a ubiquitous chloroplast endoribonuclease belonging to the short chain dehydrogenase/reductase (SDR) superfamily. To help elucidate the role of CSP41 in chloroplast gene regulation, the mechanisms that determine its substrate recognition and catalytic activity were investigated. A divalent metal is required for catalysis, most probably to provide a nucleophile for cleavage 5' to the phosphodiester bond, and may also participate in cleavage site selection. This requirement distinguishes CSP41 from other Rossman fold-containing proteins from the SDR superfamily, including several RNA-binding proteins and endonucleases. CSP41 is active only in the presence of MgCl2 and CaCl2. Although Mg2+- and Ca2+-activated CSP41 cleave at identical sites in the single-stranded regions of a stem-loop-containing substrate, Mg2+-activated CSP41 was also able to cleave within the double-stranded region of the stem-loop. Mixed metal experiments with Mg2+ and Ca2+ suggest that CSP41 contains a single divalent metal-binding site which is non-selective, since Mn2+, Co2+ and Zn2+ compete with Mg2+ for binding, although there is no activity in their presence. Using site directed mutagenesis, we identified three residues, Asn71, Asp89 and Asp103, which may form the divalent metal-binding pocket. The activation constant for Mg2+ (K(A,Mg) = 2.1 +/- 0.4 mM) is of the same order of magnitude as the stromal Mg2+ concentrations, which fluctuate between 0.5 and 10 mM as a function of light and of leaf development. These changes in stromal Mg2+ concentration may regulate CSP41 activity, and thus cpRNA stability, during plant development. PMID- 12888491 TI - Solution structure of the HIV-1 frameshift inducing stem-loop RNA. AB - The translation of reverse transcriptase and other essential viral proteins from the HIV-1 Pol mRNA requires a programmed -1 ribosomal frameshift. This frameshift is induced by two highly conserved elements within the HIV-1 mRNA: a slippery sequence comprised of a UUUUUUA heptamer, and a downstream stem-loop structure. We have determined the structure of the HIV-1 frameshift inducing RNA stem-loop, using multidimensional heteronuclear nuclear magnetic resonance (NMR) methods. The 22 nucleotide RNA solution structure [root mean squared deviation (r.m.s.d.) = 1.2 A] was determined from 475 nuclear Overhauser effect (NOE)-derived distance restrains, 20 residual dipolar couplings and direct detection of hydrogen bonds via scalar couplings. We find that the frameshift inducing stem-loop is an A-form helix capped by a structured ACAA tetraloop. The ACAA tetraloop is stabilized by an equilateral 5' and 3' stacking pattern, a sheared A-A pair and a cross-strand hydrogen bond. Unexpectedly, the ACAA tetraloop structure is nearly identical to a known tetraloop fold, previously identified in the RNase III recognition site from Saccharomyces cerevisiae. PMID- 12888493 TI - Transposases are responsible for the target specificity of IS1397 and ISKpn1 for two different types of palindromic units (PUs). AB - Insertion sequences (IS)1397 and ISKpn1, found in Escherichia coli and Klebsiella pneumoniae, respectively, are IS3 family members that insert specifically into short palindromic repeated sequences (palindromic units or PUs). In this paper, we first show that although PUs are naturally absent from extrachromosomal elements, both ISs are able to transpose from the chromosome or from a plasmid into PUs artificially introduced into target plasmids. We also show that ISKpn1 target specificity is restricted to K.pneumoniae Z1 PU type, whereas IS1397 target specificity is less stringent since the IS targets the three E.coli Y, Z1 and Z2 PU types indifferently. Experiments of transposition of both ISs driven by both transposases demonstrate that the inverted repeats flanking the ISs are not responsible for this target specificity, which is entirely due to the transposase itself. Implications on ISs evolution are presented. PMID- 12888492 TI - Fep1 represses expression of the fission yeast Schizosaccharomyces pombe siderophore-iron transport system. AB - When iron repletes, Schizosaccharomyces pombe cells repress transcription of genes encoding components involved in the reductive iron transport system. Fep1 mediates this transcriptional control by interacting specifically with GATA-type cis-acting elements. To further investigate the role that Fep1 plays in iron homeostasis, we searched for additional Fep1-regulated genes. We found that str1+ is subject to negative transcriptional regulation, which is exerted through binding of Fep1 to a single GATA element in the str1+ promoter. Introduction of str1+ into a Saccharomyces cerevisiae fet3Delta arn1-4Delta strain led to assimilation of iron from ferrichrome, revealing that Str1 functions as a siderophore-iron transporter in S.pombe. We also identified two additional target genes of Fep1, named str2+ and str3+. We demonstrate that the str1+, str2+ and str3+ genes share a common promoter element, 5'-(A/T)GATAA-3'. We found that the N-terminal 241 residue segment of Fep1 expressed in Escherichia coli specifically interacts with the 5'-(A/T)GATAA-3' element present in each of these promoters. Consistent with this, constitutive high level str1+, str2+ and str3+ gene expression was observed in a fep1Delta mutant strain. Taken together, these results demonstrate that Fep1 occupies a central role in coordinating transcriptional regulation of genes encoding components of the reductive and non reductive iron transport systems in fission yeast. PMID- 12888494 TI - Intracellular mRNA cleavage by 3' tRNase under the direction of 2'-O-methyl RNA heptamers. AB - Mammalian tRNA 3' processing endoribonuclease (3'-tRNase) can cleave any RNA at any site under the direction of small guide RNA (sgRNA) in vitro. sgRNAs can be as short as heptamers, which are much smaller than small interfering RNAs of approximately 21 nt. Together with such flexibility in substrate recognition, the ubiquity and the constitutive expression of 3'-tRNase have suggested that this enzyme can be utilized for specific cleavage of cellular RNAs by introducing appropriate sgRNAs into living cells. Here we demonstrated that the expression of chloramphenicol acetyltransferase can be downregulated by an appropriate sgRNA which is introduced into Madin-Darby canine kidney epithelial cells as an expression plasmid or a synthetic 2'-O-methyl RNA. We also showed that 2'-O methyl RNA heptamers can attack luciferase mRNAs with a high specificity and induce 3'-tRNase-mediated knock-down of the mRNAs in 293 cells. Furthermore, the MTT cell viability assay suggested that an RNA heptamer can downregulate the endogenous Bcl-2 mRNA in Sarcoma 180 cells. This novel sgRNA/3'-tRNase strategy for destroying specific cellular RNAs may be utilized for therapeutic applications. PMID- 12888495 TI - Mutagenesis of a light-regulated psbA intron reveals the importance of efficient splicing for photosynthetic growth. AB - The chloroplast-encoded psbA gene encodes the D1 polypeptide of the photosystem II reaction center, which is synthesized at high rates in the light. In Chlamydomonas reinhardtii, the psbA gene contains four self-splicing group I introns whose rates of splicing in vivo are increased at least 6-10-fold by light. However, because psbA is an abundant mRNA, and some chloroplast mRNAs appear to be in great excess of what is needed to sustain translation rates, the developmental significance of light-promoted splicing has not been clear. To address this and other questions, potentially destabilizing substitutions were made in several predicted helices of the fourth psbA intron, Cr.psbA4, and their effects on in vitro and in vivo splicing assessed. Two-nucleotide substitutions in P4 and P7 were necessary to substantially reduce splicing of this intron in vivo, although most mutations reduced self-splicing in vitro. The P7-4,5 mutant, whose splicing was completely blocked, showed no photoautotrophic growth and synthesis of a truncated D1 (exons 1-4) polypeptide from the unspliced mRNA. Most informative was the P4'-3,4 mutant, which exhibited a 45% reduction in spliced psbA mRNA, a 28% reduction in synthesis of full-length D1, and an 18% reduction in photoautotrophic growth. These results indicate that psbA mRNA is not in great excess, and that highly efficient splicing of psbA introns, which is afforded by light conditions, is necessary for optimal photosynthetic growth. PMID- 12888497 TI - The human genome contains many types of chimeric retrogenes generated through in vivo RNA recombination. AB - L1 retrotransposons play an important role in mammalian genome shaping. In particular, they can transduce their 3'-flanking regions to new genomic loci or produce pseudogenes or retrotranscripts through reverse transcription of different kinds of cellular RNAs. Recently, we found in the human genome an unusual family of chimeric retrotranscripts composed of full-sized copies of U6 small nuclear RNAs fused at their 3' termini with 5'-truncated, 3'-poly(A)-tailed L1s. The chimeras were flanked by 11-21 bp long direct repeats, and contained near their 5' ends T2A4 hexanucleotide motifs, preferably recognized by L1 nicking endonuclease. These features suggest that the chimeras were formed using the L1 integration machinery. Here we report the identification of 81 chimeras consisting of fused DNA copies of different RNAs, including mRNAs of known human genes. Based on their structural features, the chimeras were subdivided into nine distinct families. 5' Parts of the chimeras usually originated from different nuclear RNAs, whereas their 3' parts represented cytoplasmic RNAs: mRNAs, including L1 mRNA and Alu RNA. Some of these chimeric retrotranscripts are expressed in a variety of human tissues. These findings suggest that RNA-RNA recombination during L1 reverse transcription followed by the integration of the recombinants into the host genome is a general event in genome evolution. PMID- 12888496 TI - Yeast recombination pathways triggered by topoisomerase II-mediated DNA breaks. AB - Topoisomerase II is a ubiquitous enzyme that removes knots and tangles from the genetic material by generating transient double-strand DNA breaks. While the enzyme cannot perform its essential cellular functions without cleaving DNA, this scission activity is inherently dangerous to chromosomal integrity. In fact, etoposide and other clinically important anticancer drugs kill cells by increasing levels of topoisomerase II-mediated DNA breaks. Cells rely heavily on recombination to repair double-strand DNA breaks, but the specific pathways used to repair topoisomerase II-generated DNA damage have not been defined. Therefore, Saccharomyces cerevisiae was used as a model system to delineate the recombination pathways that repair DNA breaks generated by topoisomerase II. Yeast cells that expressed wild-type or a drug-hypersensitive mutant topoisomerase II or overexpressed the wild-type enzyme were examined. Based on cytotoxicity and recombination induced by etoposide in different repair-deficient genetic backgrounds, double-strand DNA breaks generated by topoisomerase II appear to be repaired primarily by the single-strand invasion pathway of homologous recombination. Non-homologous end joining also was triggered by etoposide treatment, but this pathway was considerably less active than single strand invasion and did not contribute significantly to cell survival in S.cerevisiae. PMID- 12888498 TI - Structural and functional homology between the RNAP(I) subunits A14/A43 and the archaeal RNAP subunits E/F. AB - In the archaeal RNA polymerase and the eukaryotic RNA polymerase II, two subunits (E/F and RPB4/RPB7, respectively) form a heterodimer that reversibly associates with the core of the enzyme. Recently it has emerged that this heterodimer also has a counterpart in the other eukaryotic RNA polymerases: in particular two subunits of RNA polymerase I (A14 and A43) display genetic and biochemical characteristics that are similar to those of the RPB4 and RPB7 subunits, despite the fact that only A43 shows some sequence homology to RPB7. We demonstrate that the sequence of A14 strongly suggests the presence of a HRDC domain, a motif that is found at the C-terminus of a number of helicases and RNases. The same motif is also seen in the structure of the F subunit, suggesting a structural link between A14 and the RPB4/C17/subunit F family, even in the absence of direct sequence homology. We show that it is possible to co-express and co-purify large amounts of the recombinant A14/A43 heterodimer, indicating a tight and specific interaction between the two subunits. To shed light on the function of the heterodimer, we performed gel mobility shift assays and showed that the A14/A43 heterodimer binds single-stranded RNA in a similar way to the archaeal E/F complex. PMID- 12888499 TI - Structural diversification and neo-functionalization during floral MADS-box gene evolution by C-terminal frameshift mutations. AB - Frameshift mutations generally result in loss-of-function changes since they drastically alter the protein sequence downstream of the frameshift site, besides creating premature stop codons. Here we present data suggesting that frameshift mutations in the C-terminal domain of specific ancestral MADS-box genes may have contributed to the structural and functional divergence of the MADS-box gene family. We have identified putative frameshift mutations in the conserved C terminal motifs of the B-function DEF/AP3 subfamily, the A-function SQUA/AP1 subfamily and the E-function AGL2 subfamily, which are all involved in the specification of organ identity during flower development. The newly evolved C terminal motifs are highly conserved, suggesting a de novo generation of functionality. Interestingly, since the new C-terminal motifs in the A- and B function subfamilies are only found in higher eudicotyledonous flowering plants, the emergence of these two C-terminal changes coincides with the origin of a highly standardized floral structure. We speculate that the frameshift mutations described here are examples of co-evolution of the different components of a single transcription factor complex. 3' terminal frameshift mutations might provide an important but so far unrecognized mechanism to generate novel functional C-terminal motifs instrumental to the functional diversification of transcription factor families. PMID- 12888500 TI - Hydropathic analysis of the free energy differences in anthracycline antibiotic binding to DNA. AB - Molecular models of six anthracycline antibiotics and their complexes with 32 distinct DNA octamer sequences were created and analyzed using HINT (Hydropathic INTeractions) to describe binding. The averaged binding scores were then used to calculate the free energies of binding for comparison with experimentally determined values. In parsing our results based on specific functional groups of doxorubicin, our calculations predict a free energy contribution of -3.6 +/- 1.1 kcal x mol(-1) (experimental -2.5 +/- 0.5 kcal x mol(-1)) from the groove binding daunosamine sugar. The net energetic contribution of removing the hydroxyl at position C9 is -0.7 +/- 0.7 kcal x mol(-1) (-1.1 +/- 0.5 kcal x mol(-1)). The energetic contribution of the 3' amino group in the daunosamine sugar (when replaced with a hydroxyl group) is -3.7 +/- 1.1 kcal x mol(-1) (-0.7 +/- 0.5 kcal x mol(-1)). We propose that this large discrepancy may be due to uncertainty in the exact protonation state of the amine. The energetic contribution of the hydroxyl group at C14 is +0.4 +/- 0.6 kcal x mol(-1) (-0.9 +/- 0.5 kcal x mol( 1)), largely due to unfavorable hydrophobic interactions between the hydroxyl oxygen and the methylene groups of the phosphate backbone of the DNA. Also, there appears to be considerable conformational uncertainty in this region. This computational procedure calibrates our methodology for future analyses where experimental data are unavailable. PMID- 12888501 TI - The activity of siRNA in mammalian cells is related to structural target accessibility: a comparison with antisense oligonucleotides. AB - The biological activity of siRNA seems to be influenced by local characteristics of the target RNA, including local RNA folding. Here, we investigated quantitatively the relationship between local target accessibility and the extent of inhibition of the target gene by siRNA. Target accessibility was assessed by a computational approach that had been shown earlier to be consistent with experimental probing of target RNA. Two sites of ICAM-1 mRNA predicted to serve as accessible motifs and one site predicted to adopt an inaccessible structure were chosen to test siRNA constructs for suppression of ICAM-1 gene expression in ECV304 cells. The local target-dependent effectiveness of siRNA was compared with antisense oligonucleotides (asON). The concentration dependency of siRNA-mediated suppression indicates a >1000-fold difference between active siRNAs (IC50 approximately 0.2-0.5 nM) versus an inactive siRNA (IC50 > or = 1 microM) which is consistent with the activity pattern of asON when relating target suppression to predicted local target accessibility. The extremely high activity of the siRNA si2B (IC50 = 0.24 nM) indicates that not all siRNAs shown to be active at the usual concentrations of >10-100 nM belong to this highly active species. The observations described here suggest an option to assess target accessibility for siRNA and, thus, support the design of active siRNA constructs. This approach can be automated, work at high throughput and is open to include additional parameters relevant to the biological activity of siRNA. PMID- 12888502 TI - Spurious spatial periodicity of co-expression in microarray data due to printing design. AB - Global transcriptome data is increasingly combined with sophisticated mathematical analyses to extract information about the functional state of a cell. Yet the extent to which the results reflect experimental bias at the expense of true biological information remains largely unknown. Here we show that the spatial arrangement of probes on microarrays and the particulars of the printing procedure significantly affect the log-ratio data of mRNA expression levels measured during the Saccharomyces cerevisiae cell cycle. We present a numerical method that filters out these technology-derived contributions from the existing transcriptome data, leading to improved functional predictions. The example presented here underlines the need to routinely search and compensate for inherent experimental bias when analyzing systematically collected, internally consistent biological data sets. PMID- 12888503 TI - DNA polymerase III chi subunit ties single-stranded DNA binding protein to the bacterial replication machinery. AB - Single-stranded DNA binding (SSB) protein binds to single-stranded DNA (ssDNA) at the lagging strand of the replication fork in Escherichia coli cells. This protein is essential for the survival of the E.coli cell, presumably because it shields the ssDNA and holds it in a suitable conformation for replication by DNA polymerase III. In this study we undertook a biophysical analysis of the interaction between the SSB protein of E.coli and the chi subunit of DNA polymerase III. Using analytical ultracentrifugation we show that at low salt concentrations there is an increase in the stability in the physical interaction between chi and an EcoSSB/ssDNA complex when compared to that of chi to EcoSSB alone. This increase in stability disappeared in high salt conditions. The sedimentation of an EcoSSB protein lacking its C-terminal 26 amino acids remains unchanged in the presence of chi, showing that chi interacts specifically with the C-terminus of EcoSSB. In DNA melting experiments we demonstrate that chi specifically enhances the ssDNA stabilization by EcoSSB. Thus, the binding of EcoSSB to chi at the replication fork prevents premature dissociation of EcoSSB from the lagging strand and thereby enhances the processivity of DNA polymerase III. PMID- 12888504 TI - Lack of sugar discrimination by human Pol mu requires a single glycine residue. AB - DNA polymerase mu (Pol mu) is a novel family X DNA polymerase that has been suggested to play a role in micro-homology mediated joining and repair of double strand breaks. We show here that human Pol mu is not able to discriminate against the 2'-OH group of the sugar moiety. It inserts rNTPs with an efficiency that is <10-fold lower than that of dNTPs, in sharp contrast with the >1000-fold discrimination characteristic of most DNA-dependent DNA polymerases. The lack of sugar discrimination by Pol mu is demonstrated by its ability to add rNTPs to both DNA and RNA primer strands, and to insert both deoxy- and ribonucleotides on growing nucleic acid chains. 3D-modelling of human Pol mu based on the available Pol beta and TdT structural information allowed us to predict candidate residues involved in sugar discrimination. Thus, a single amino acid substitution in which Gly433 residue of Pol mu was mutated to the consensus tyrosine present in Pol beta, produced a strong increase in the discrimination against ribonucleotides. The unusual capacity to insert both rNTPs and dNTPs will be discussed in the context of the predicted roles of Pol mu in DNA repair. PMID- 12888505 TI - Microenvironment analysis and identification of magnesium binding sites in RNA. AB - Interactions with magnesium (Mg2+) ions are essential for RNA folding and function. The locations and function of bound Mg2+ ions are difficult to characterize both experimentally and computationally. In particular, the P456 domain of the Tetrahymena thermophila group I intron, and a 58 nt 23s rRNA from Escherichia coli have been important systems for studying the role of Mg2+ binding in RNA, but characteristics of all the binding sites remain unclear. We therefore investigated the Mg2+ binding capabilities of these RNA systems using a computational approach to identify and further characterize their Mg2+ binding sites. The approach is based on the FEATURE algorithm, reported previously for microenvironment analysis of protein functional sites. We have determined novel physicochemical descriptions of site-bound and diffusely bound Mg2+ ions in RNA that are useful for prediction. Electrostatic calculations using the Non-Linear Poisson Boltzmann (NLPB) equation provided further evidence for the locations of site-bound ions. We confirmed the locations of experimentally determined sites and further differentiated between classes of ion binding. We also identified potentially important, high scoring sites in the group I intron that are not currently annotated as Mg2+ binding sites. We note their potential function and believe they deserve experimental follow-up. PMID- 12888506 TI - The synthesis of oligoribonucleotides containing N6-alkyladenosines and 2 methylthio-N6-alkyladenosines via post-synthetic modification of precursor oligomers. AB - The N6-alkyladenosines and 2-methylthio-N6-alkyladenosines are the most common modified adenosine nucleosides and transfer ribonucleic acids (tRNA) are particularly rich in these modified nucleosides. They are present at position 37 of the anticodon arm and the contribution of these hypermodified nucleosides to codon-anticodon interactions, as well as translation, are significant, although not fully understood. Herein we described a new chemical synthesis method of the oligoribonucleotides containing N6-alkyladenosines and 2-methylthio-N6 alkyladenosines via post-synthetic modifications of precursor oligoribonucleotides. To obtain oligoribonucleotides containing N6 alkyladenosines, the precursor oligoribonucleotide carrying 6-methylthiopurine riboside residue was used, whereas for the synthesis of oligoribonucleotides containing 2-methylthio-N6-alkyladenosines the precursor oligoribonucleotide carrying the 2-methylthio-6-chloropurine riboside was applied. Among the modified oligoribonucleotides of different length and secondary structures, there were several containing naturally occurring modified nucleosides such as: N6 isopentenyladenosine (i6A), N6-methyladenosine (m6A), 2-methylthio-N6 isopentenyladenosine (ms2i6A), and 2-methylthio-N6-methyladenosine (ms2m6A), as well as several unnaturally modified adenosine derivatives. PMID- 12888507 TI - The thermodynamic stability of RNA duplexes and hairpins containing N6 alkyladenosines and 2-methylthio-N6-alkyladenosines. AB - The N6-alkyladenosines and 2-methylthio-N6-alkyladenosines make up over half of the population of all naturally modified adenosines and they are present in the transfer ribonucleic acids (tRNA) at position 37. We measured effects of N6 alkyladenosines and 2-methylthio-N6-alkyladenosines on the thermodynamic stability of RNA duplexes containing a U-A(Mod) base pair at internal and terminal duplex positions, as well as containing modified adenosines as a 3' terminal unpaired nucleotide. Beside naturally modified adenosines such as N6 isopentenyladenosine (i6A), N6-methyladenosine (m6A), 2-methylthio-N6 isopentenyladenosine (ms2i6A) and 2-methylthio-N6-methyladenosine (ms2m6A), we studied several artificial modifications to evaluate the steric and electronic effects of N6-alkyl substituents. Moreover, some N6-alkyladenosines and 2 methylthio-N6-alkyladenosines were placed in hairpins at positions corresponding to nucleotide 37 of the tRNA anticodon arm, and the thermodynamic stability of those hairpins was studied. The stability of the modified RNA hairpins was measured in standard melting buffer containing 1 M sodium chloride as well as in physiological buffer containing 10 mM magnesium chloride and 150 mM potassium chloride. The results obtained indicate that the nature of the adenosine modification and the position of U-A(Mod) base pairs within the duplex influence the thermodynamic stability of RNA duplexes. For most of the modification, the destabilization of duplexes was observed. Moreover, we found that the buffer composition and the structure of the modified adenosine very significantly affect the thermodynamic stability of RNA. PMID- 12888508 TI - Double-stranded RNA-mediated gene silencing in fission yeast. AB - Double-stranded RNA (dsRNA) can specifically inhibit gene expression in a variety of organisms by invoking post-transcriptional degradation of homologous mRNA. Here we show that dsRNA-mediated gene regulation also occurs in the fission yeast Schizosaccharomyces pombe. We present evidence that: (i) reporter gene silencing is significantly enhanced when additional non-coding sense RNA is co-expressed with antisense RNA; (ii) expression of a panhandle RNA also silences target gene expression; (iii) expression of dsRNA is associated with siRNAs; (iv) a novel host-encoded factor which enhances antisense RNA gene silencing also enhances panhandle RNA-mediated gene inhibition. Both the exogenously introduced lacZ and c-myc genes are shown to be susceptible to dsRNA- mediated gene silencing in this model. Taken together, these data indicate that RNA-mediated gene silencing can occur through a RNAi-like mechanism in fission yeast. PMID- 12888509 TI - Global methylation screening in the Arabidopsis thaliana and Mus musculus genome: applications of virtual image restriction landmark genomic scanning (Vi-RLGS). AB - Understanding the role of 'epigenetic' changes such as DNA methylation and chromatin remodeling has now become critical in understanding many biological processes. In order to delineate the global methylation pattern in a given genomic DNA, computer software has been developed to create a virtual image of restriction landmark genomic scanning (Vi-RLGS). When using a methylation- sensitive enzyme such as NotI as the restriction landmark, the comparison between real and in silico RLGS profiles of the genome provides a methylation map of genomic NotI sites. A methylation map of the Arabidopsis genome was created that could be confirmed by a methylation-sensitive PCR assay. The method has also been applied to the mouse genome. Although a complete methylation map has not been completed, a region of methylation difference between two tissues has been tested and confirmed by bisulfite sequencing. Vi-RLGS in conjunction with real RLGS will make it possible to develop a more complete map of genomic sites that are methylated or demethylated as a consequence of normal or abnormal development. PMID- 12888510 TI - Oligoamine-acridine conjugates for promotion of gap-selective DNA hydrolysis by Ce(IV)/EDTA complex. AB - Oligoamines (spermidine, dipropylenetriamine and propylenediamine) were covalently attached to acridine via a hexamethylene linker. These oligoamine acridine conjugates were efficiently bound to gap sites in substrate DNA, and promoted the DNA hydrolysis by a homogeneous Ce(IV)/ethylenediamine-N,N,N',N' tetraacetate (EDTA) complex at these sites. In contrast, the hydrolysis of the double-stranded portion in the DNA was little affected by these conjugates, although they were strongly bound thereto by the intercalation of their acridine moieties. As a result, the gap site was selectively and efficiently hydrolyzed by combining the Ce(IV)/EDTA complex with the oligoamine--acridine conjugate. Either the oligoamine or the acridine was only poorly active for the purpose, substantiating the essential role of cooperation between them. The promotion of gap-selective DNA hydrolysis by the conjugates has been ascribed to electrostatic stabilization of a negatively charged transition state by their positive charges. PMID- 12888511 TI - Evolutionary clues to DNA polymerase III beta clamp structural mechanisms. AB - The prokaryotic DNA polymerase III beta homodimeric clamp links the replication complex to DNA during polynucleotide synthesis. This clamp is loaded onto DNA and unloaded by the clamp loader complex, the delta subunit of which by itself can bind to and open the clamp. beta Clamps from diverse bacteria were examined using contrast hierarchical alignment and interaction network (CHAIN) analysis, a statistical approach that categorizes and measures the evolutionary constraints imposed on protein sequences by natural selection. Some constraints are subtle inasmuch as they are unique to certain bacteria. Examination of corresponding molecular interactions within structures of the Escherichia coli beta dimeric and delta-beta complexes reveals that N320, Y323 and R176, which are subject to very strong constraints, form a substructure that may serve as a platform for leveraging and directing delta-induced conformational changes. N320 may play a prominent role, as it is strategically situated between this substructure and regions linked to delta binding and opening of beta's dimeric interface. R176 appears to act as a relay between the delta binding site and the clamp's central hole. Other residues subject to strong constraints are likewise associated with structurally important features. For example, two pairs of interacting residues, R269/E304 and K74/E300, form salt bridges at the dimeric interface, while the C terminal residues M362, P363, M364 and R365 appear to play key roles in delta binding. Q149 and K198 appear to sense DNA within the clamp's central hole while other residues may relay this information to the delta binding site. Mutagenesis experiments designed to explore possible mechanisms are proposed. PMID- 12888512 TI - Transcription increases multiple spontaneous point mutations in Salmonella enterica. AB - The spontaneous rate of G.C-->A.T mutations and a hotspot T.A-->G.C transversion are known to increase with the frequency of transcription-increases that have been ascribed primarily to processes that affect only these specific mutations. To investigate how transcription induces other spontaneous point mutations, we tested for its effects in repair-proficient Salmonella enterica using reversion assays of chromosomally inserted alleles. Our results indicate that transcription increases rates of all tested point mutations in the induced gene: induction significantly increased the individual rates of an A.T-->T.A transversion, an A.T ->G.C transition and the pooled rates of the three other point mutations assayed. Although the S.enterica genome is thought to have a mutational bias towards G.C base pairs, transitions creating A.T pairs were approximately 10 times more frequent than the reverse mutation, resulting in an overall mutation pressure to lower G+C contents. Transitions occurred at roughly twice the rate of transversions, similar to results from sequence comparisons; however, several individual transversions are more frequent than the least common transition. PMID- 12888513 TI - Low pH enhances Sp1 DNA binding activity and interaction with TBP. AB - Sp1 is involved in the regulation of a wide variety of genes, including housekeeping genes and genes involved in tumor growth. Sp1 is a member of the C2 H2 zinc-finger family and is important for protection against cellular acidosis in cells that grow under hypoxic or acidic conditions, such as tumor cells. To obtain an insight into the molecular mechanisms underlying pH-dependent transcription by Sp1, both its DNA binding activity and its interaction with TATA binding protein (TBP) were investigated under various pH conditions. We show here that the DNA binding activity of Sp1 increased and Sp1 formed a stable interaction with TBP at low pH. These findings indicate that pH changes significantly modulate the activity of Sp1 and thus contribute to the cellular response under hypoxic or acidic conditions. PMID- 12888514 TI - Patterns of sequence conservation at termini of long terminal repeat (LTR) retrotransposons and DNA transposons in the human genome: lessons from phage Mu. AB - Long terminal repeat (LTR) retrotransposons and DNA transposons are transposable elements (TEs) that perform cleavage and transfer at precise DNA positions. Here, we present statistical analyses of sequences found at the termini of precise TEs in the human genome. The results show that the terminal di- and trinucleotides of these TEs are highly conserved. 5'TG...CA3' occurs most frequently at the termini of LTR retrotransposons, while 5'CAG...CTG3' occurs most frequently in DNA transposons. Interestingly, these sequences are the most flexible base pair steps in DNA. Both the sequence preference and the degree of conservation of each position within the human LTR dinucleotide termini are remarkably similar to those experimentally demonstrated in transposable phage Mu. We discuss the significance of these observations and their implication for the function of terminal residues in the transposition of precise TEs. PMID- 12888515 TI - Roles of Saccharomyces cerevisiae DNA polymerases Poleta and Polzeta in response to irradiation by simulated sunlight. AB - Sunlight causes lesions in DNA that if unrepaired and inaccurately replicated by DNA polymerases yield mutations that result in skin cancer in humans. Two enzymes involved in translesion synthesis (TLS) of UV-induced photolesions are DNA polymerase eta (Poleta) and polymerase zeta (Polzeta), encoded by the RAD30A and REV3 genes, respectively. Previous studies have investigated the TLS roles of these polymerases in human and yeast cells irradiated with monochromatic, short wavelength UVC radiation (254 nm). However, less is known about cellular responses to solar radiation, which is of higher and mixed wavelengths (310-1100 nm) and produces a different spectrum of DNA lesions, including Dewar photoproducts and oxidative lesions. Here we report on the comparative cytotoxic and mutagenic effects of simulated sunlight (SSL) and UVC radiation on yeast wild type, rad30Delta, rev3Delta and rev3Delta rad30Delta strains. The results with SSL support several previous interpretations on the roles of these two polymerases in TLS of photodimers and (6-4) photoproducts derived from studies with UVC. They further suggest that Poleta participates in the non-mutagenic bypass of SSL-dependent cytosine-containing Dewar photoproducts and 8-oxoguanine, while Polzeta is mainly responsible for the mutagenic bypass of all types of Dewar photoproducts. They also suggest that in the absence of Polzeta, Poleta contributes to UVC- and SSL-induced mutagenesis, possibly by the bypass of photodimers containing deaminated cytosine. PMID- 12888516 TI - The computational analysis of scientific literature to define and recognize gene expression clusters. AB - A limitation of many gene expression analytic approaches is that they do not incorporate comprehensive background knowledge about the genes into the analysis. We present a computational method that leverages the peer-reviewed literature in the automatic analysis of gene expression data sets. Including the literature in the analysis of gene expression data offers an opportunity to incorporate functional information about the genes when defining expression clusters. We have created a method that associates gene expression profiles with known biological functions. Our method has two steps. First, we apply hierarchical clustering to the given gene expression data set. Secondly, we use text from abstracts about genes to (i) resolve hierarchical cluster boundaries to optimize the functional coherence of the clusters and (ii) recognize those clusters that are most functionally coherent. In the case where a gene has not been investigated and therefore lacks primary literature, articles about well-studied homologous genes are added as references. We apply our method to two large gene expression data sets with different properties. The first contains measurements for a subset of well-studied Saccharomyces cerevisiae genes with multiple literature references, and the second contains newly discovered genes in Drosophila melanogaster; many have no literature references at all. In both cases, we are able to rapidly define and identify the biologically relevant gene expression profiles without manual intervention. In both cases, we identified novel clusters that were not noted by the original investigators. PMID- 12888517 TI - Identification, characterization and molecular phylogeny of U12-dependent introns in the Arabidopsis thaliana genome. AB - U12-dependent introns are spliced by the minor U12-type spliceosome and occur in a variety of eukaryotic organisms, including Arabidopsis. In this study, a set of putative U12-dependent introns was compiled from a large collection of cDNA/EST- confirmed introns in the Arabidopsis thaliana genome by means of high-throughput bioinformatic analysis combined with manual scrutiny. A total of 165 U12-type introns were identified based upon stringent criteria. This number of sequences well exceeds the total number of U12-type introns previously reported for plants and allows a more thorough statistical analysis of U12-type signals. Of particular note is the discovery that the distance between the branch site adenosine and the acceptor site ranges from 10 to 39 nt, significantly longer than the previously postulated limit of 21 bp. Further analysis indicates that, in addition to the spacing constraint, the sequence context of the potential acceptor site may have an important role in 3' splice site selection. Several alternative splicing events involving U12-type introns were also captured in this study, providing evidence that U12-dependent acceptor sites can also be recognized by the U2-type spliceosome. Furthermore, phylogenetic analysis suggests that both U12-type AT-AC and U12-type GT-AG introns occurred in Na+/H+ antiporters in a progenitor of animals and plants. PMID- 12888518 TI - Repair of clustered uracil DNA damages in Escherichia coli. AB - Multiply damaged sites (MDS) are defined as greater than/equal to two lesions within 10-15 bp and are generated in DNA by ionizing radiation. In vitro repair of closely opposed base damages > or =2 bp apart results in a double strand break (DSB). This work extends the in vitro studies by utilizing clusters of uracil DNA damage as model lesions to determine whether MDS are converted to DSBs in bacteria. Lesions were positioned within the firefly luciferase coding region, transformed into bacteria (wild-type, uracil DNA glycosylase-deficient, ung-, or exonuclease III and endonuclease IV-deficient, xth-nfo-) and luciferase activity measured following repair. DSB formation was expected to decrease activity. Two closely opposed uracils separated by < or =7 bp decreased luciferase activity in wild-type and xth-nfo-, but not ung- bacteria. Growth of bacteria to obtain plasmid-containing colonies demonstrated that the plasmid was destroyed following the mis-repair of two uracils positioned 7 bp apart. This study indicates a DSB is formed when uracil DNA glycosylase initiates repair of two closely opposed uracils < or =7 bp apart, even in the absence of the major apurinic endonucleases. This work supports the in vitro studies and demonstrates that DNA repair is not always advantageous to cells. PMID- 12888519 TI - The ERV-9 LTR enhancer is not blocked by the HS5 insulator and synthesizes through the HS5 site non-coding, long RNAs that regulate LTR enhancer function. AB - A solitary long terminal repeat (LTR) of ERV-9 human endogenous retrovirus is located upstream of the HS5 site in the human beta-globin locus control region and possesses unique enhancer activity in erythroid K562 cells. In cells transfected with plasmid LTR-HS5-epsilonp-GFP, the LTR enhancer activates the GFP reporter gene and is not blocked by the interposed HS5 site, which has been reported to have insulator function. The LTR enhancer initiates synthesis of long RNAs from the LTR promoter through the intervening HS5 site into the epsilon globin promoter and the GFP gene. Synthesis of the sense, long LTR RNAs is correlated with high level synthesis of GFP mRNA from the epsilon-globin promoter. Mutations of the LTR promoter and/or the epsilon-globin promoter show that (i) the LTR enhancer can autonomously initiate synthesis of LTR RNAs independent of the promoters and (ii) the LTR RNAs are not processed into GFP mRNA or translated into GFP. However, reversing the orientation of the LTR in plasmid (LTR)rev-HS5-epsilonp-GFP, thus reversing the direction of synthesis of LTR RNAs in the antisense direction away from the epsilon-globin promoter and GFP gene drastically reduces the level of GFP mRNA and thus LTR enhancer function. The results suggest that the LTR-assembled transcription machinery in synthesizing non-coding, LTR RNAs can reach the downstream epsilon-globin promoter to activate transcription of the GFP gene. PMID- 12888520 TI - The essential transcription factor Reb1p interacts with the CLB2 UAS outside of the G2/M control region. AB - Regulation of CLB2 is important both for completion of the normal vegetative cell cycle in Saccharomyces cerevisiae and for departure from the vegetative cell cycle upon nitrogen deprivation. Cell cycle-regulated transcription of CLB2 in the G2/M phase is known to be brought about by a set of proteins including Mcm1p, Fkh2/1p and Ndd1p that associate with a 35 bp G2/M-specific sequence common to a set of co-regulated genes. CLB2 transcription is regulated by additional signals, including by nitrogen levels, by positive feedback from the Clb2-Cdc28 kinase, and by osmotic stress, but the corresponding regulatory sequences and proteins have not been identified. We have found that the essential Reb1 transcription factor binds with high affinity to a sequence upstream of CLB2, within a region implicated previously by others in regulated expression, but upstream of the known G2/M-specific site. CLB2 sequence from the region around the Reb1p site blocks activation by the Gal4 protein when positioned downstream of the Gal4 binding site. Since a mutation in the Reb1p site abrogates this effect, we suggest that Reb1p is likely to occupy this site in vivo. PMID- 12888521 TI - Controlling nucleic acid secondary structure by intercalation: effects of DNA strand length on coralyne-driven duplex disproportionation. AB - Small molecules that intercalate in DNA and RNA are powerful agents for controlling nucleic acid structural transitions. We recently demonstrated that coralyne, a small crescent-shaped molecule, can cause the complete and irreversible disproportionation of duplex poly(dA)*poly(dT) into triplex poly(dA)*poly(dT)*poly(dT) and a poly(dA) self-structure. Both DNA secondary structures that result from duplex disproportionation are stabilized by coralyne intercalation. In the present study, we show that the kinetics and thermodynamics of coralyne-driven duplex disproportionation strongly depend on oligonucleotide length. For example, disproportionation of duplex (dA)16*(dT)16 by coralyne reverts over the course of hours if the sample is maintained at 4 degrees C. Coralyne-disproportioned (dA)32. (dT)(32), on the other hand, only partially reverts to the duplex state over the course of days at the same temperature. Furthermore, the equilibrium state of a (dA)16*(dT)16 sample in the presence of coralyne at room temperature contains three different secondary structures [i.e. duplex, triplex and the (dA)16 self-structure]. Even the well-studied process of triplex stabilization by coralyne binding is found to be a length-dependent phenomenon and more complicated than previously appreciated. Together these observations indicate that at least one secondary structure in our nucleic acid system [i.e. duplex, triplex or (dA)n self-structure] binds coralyne in a length dependent manner. PMID- 12888522 TI - The quakingviable mutation affects qkI mRNA expression specifically in myelin producing cells of the nervous system. AB - The genetic lesion in the quakingviable (qk(v)) mutant mice is a deletion 5' to the qkI gene, resulting in severe hypomyelination. qkI produces several QKI protein isoforms via alternative splicing of the C-terminal coding exons. In the qk(v)/qk(v) brain, immunostaining of QKI proteins is diminished in an isoform differential manner with undefined mechanisms. We examined the expression of QKI protein isoforms and qkI mRNA isoforms in the qk(v)/qk(v) mutants and the non phenotypic wt/qk(v) littermates. Our results indicated significant reduction of all qkI mRNA isoforms in the central and peripheral nervous system during active myelination without detectable post-transcriptional abnormalities. In the early stage of myelin development, qkI mRNAs are differentially reduced, which appeared to be responsible for the reduction of the corresponding QKI protein isoforms. The reduced qkI expression was a specific consequence of the qk(v) lesion, not observed in other hypomyelination mutants. Further more, no abnormal qkI expression was found in testis, heart and astroglia of the qk(v)/qk(v) mice, suggesting that the reduction of qkI mRNAs occurred specifically in myelin producing cells of the nervous system. These observations suggest that diminished qkI expression results from deletion of an enhancer that promotes qkI transcription specifically in myelinating glia during active myelinogenesis. PMID- 12888523 TI - Structural details (kinks and non-alpha conformations) in transmembrane helices are intrahelically determined and can be predicted by sequence pattern descriptors. AB - One of the promising methods of protein structure prediction involves the use of amino acid sequence-derived patterns. Here we report on the creation of non degenerate motif descriptors derived through data mining of training sets of residues taken from the transmembrane-spanning segments of polytopic proteins. These residues correspond to short regions in which there is a deviation from the regular alpha-helical character (i.e. pi-helices, 3(10)-helices and kinks). A 'search engine' derived from these motif descriptors correctly identifies, and discriminates amongst instances of the above 'non-canonical' helical motifs contained in the SwissProt/TrEMBL database of protein primary structures. Our results suggest that deviations from alpha-helicity are encoded locally in sequence patterns only about 7-9 residues long and can be determined in silico directly from the amino acid sequence. Delineation of such variations in helical habit is critical to understanding the complex structure-function relationships of polytopic proteins and for drug discovery. The success of our current methodology foretells development of similar prediction tools capable of identifying other structural motifs from sequence alone. The method described here has been implemented and is available on the World Wide Web at http://cbcsrv.watson.ibm.com/Ttkw.html. PMID- 12888524 TI - Protein families and TRIBES in genome sequence space. AB - Accurate detection of protein families allows assignment of protein function and the analysis of functional diversity in complete genomes. Recently, we presented a novel algorithm called TribeMCL for the detection of protein families that is both accurate and efficient. This method allows family analysis to be carried out on a very large scale. Using TribeMCL, we have generated a resource called TRIBES that contains protein family information, comprising annotations, protein sequence alignments and phylogenetic distributions describing 311 257 proteins from 83 completely sequenced genomes. The analysis of at least 60 934 detected protein families reveals that, with the essential families excluded, paralogy levels are similar between prokaryotes, irrespective of genome size. The number of essential families is estimated to be between 366 and 426. We also show that the currently known space of protein families is scale free and discuss the implications of this distribution. In addition, we show that smaller families are often formed by shorter proteins and discuss the reasons for this intriguing pattern. Finally, we analyse the functional diversity of protein families in entire genome sequences. The TRIBES protein family resource is accessible at http://www.ebi.ac.uk/research/cgg/tribes/. PMID- 12888525 TI - Computational identification of protein coding potential of conserved sequence tags through cross-species evolutionary analysis. AB - The identification of conserved sequence tags (CSTs) through comparative genome analysis may reveal important regulatory elements involved in shaping the spatio temporal expression of genetic information. It is well known that the most significant fraction of CSTs observed in human-mouse comparisons correspond to protein coding exons, due to their strong evolutionary constraints. As we still do not know the complete gene inventory of the human and mouse genomes it is of the utmost importance to establish if detected conserved sequences are genes or not. We propose here a simple algorithm that, based on the observation of the specific evolutionary dynamics of coding sequences, efficiently discriminates between coding and non-coding CSTs. The application of this method may help the validation of predicted genes, the prediction of alternative splicing patterns in known and unknown genes and the definition of a dictionary of non-coding regulatory elements. PMID- 12888526 TI - Retrotransposon-mediated restoration of Chlorella telomeres: accumulation of Zepp retrotransposons at termini of newly formed minichromosomes. AB - To elucidate the contribution of LINE-like retrotransposon Zepp elements to the formation and maintenance of chromosomal telomeres, newly formed minichromosomes in irradiated Chlorella vulgaris cells were isolated and structurally characterized. A minichromosome (miniV4) of approximately 700 kb in size contained a Zepp cluster taking the place of the telomeric repeats on one terminus, whereas the other end of this chromosome consisted of canonical telomeric repeats. The Zepp copies in this cluster were in a tandem array with their poly(A) tails towards the centromere. Another minichromosome Y32 ( approximately 400 kb in size) was shown to have several copies of Zepp elements on both termini. On the right arm terminus, two copies of Zepp were found in a tandem array with poly(A) tracts facing towards the chromosomal end. The poly(A) tail and the 3'-end of approximately 400 bp of the distal copy were replaced by the telomeric repeats. On the 5'-side of the proximal copy was another Zepp element in the reverse orientation. These newly formed telomeric structures are very similar to those previously found in the left arm of chromosome I and the terminus of an unidentified chromosome and support the model of Zepp-mediated restoration and maintenance of Chlorella telomeres. PMID- 12888528 TI - Correcting errors in shotgun sequences. AB - Sequencing errors in combination with repeated regions cause major problems in shotgun sequencing, mainly due to the failure of assembly programs to distinguish single base differences between repeat copies from erroneous base calls. In this paper, a new strategy designed to correct errors in shotgun sequence data using defined nucleotide positions, DNPs, is presented. The method distinguishes single base differences from sequencing errors by analyzing multiple alignments consisting of a read and all its overlaps with other reads. The construction of multiple alignments is performed using a novel pattern matching algorithm, which takes advantage of the symmetry between indices that can be computed for similar words of the same length. This allows for rapid construction of multiple alignments, with no previous pair-wise matching of sequence reads required. Results from a C++ implementation of this method show that up to 99% of sequencing errors can be corrected, while up to 87% of the single base differences remain and up to 80% of the corrected reads contain at most one error. The results also show that the method outperforms the error correction method used in the EULER assembler. The prototype software, MisEd, is freely available from the authors for academic use. PMID- 12888527 TI - Functional similarity of Knirps CtBP-dependent and CtBP-independent transcriptional repressor activities. AB - Short-range transcriptional repressors are locally acting factors that play important roles in developmental gene expression in Drosophila. To effect repression, Knirps and other short-range repressors bind the CtBP corepressor, but these repressors also function via CtBP-independent pathways. Possible mechanistic differences between CtBP-dependent and -independent repression activities are poorly understood. The distinct activities might provide qualitatively different activities necessary in different promoter contexts, or they might combine to give quantitatively different effects. We analyze separately the CtBP-dependent and CtBP-independent domains of Knirps previously characterized in the embryo to determine possible functional distinctions of the two repression activities. Both domains are active in cell culture and are dependent on the same residues required for activity in the embryo. The domains have similar properties with respect to distance-dependent repression and resistance to inhibition by the deacetylase inhibitor trichostatin A. In tests of repressor-activator specificity, the extent of repression was related not to the chemical nature of the activation domain but to the total activation potential. This result indicates that the balance of competing activation and repression signals is decisive in determining the effectiveness of repressors on genetic switches, suggesting that multiple repression activities are utilized to provide quantitatively, rather than qualitatively, distinct outputs. PMID- 12888529 TI - Establishment of conditional vectors for hairpin siRNA knockdowns. AB - Small interference RNA (siRNA) is an emerging methodology in reverse genetics. Here we report the development of a new tetracycline-inducible vector-based siRNA system, which uses a tetracycline-responsive derivative of the U6 promoter and the tetracycline repressor for conditional in vivo transcription of short hairpin RNA. This method prevents potential lethality immediately after transfection of a vector when the targeted gene is indispensable, or the phenotype of the knockdown is lethal or results in a growth abnormality. We show that the controlled knockdown of DNA methyltransferase 1 (DNMT1) in human cancer resulted in growth arrest. Removal of the inducer, doxycycline, from treated cells led to re expression of the targeted gene. Thus the method allows for a highly controlled approach to gene knockdown. PMID- 12888530 TI - Highly stable and efficient mRNA templates for mRNA-protein fusions and C terminally labeled proteins. AB - For high-throughput in vitro protein selection using genotype (mRNA)-phenotype (protein) fusion formation and C-terminal protein labeling as a post-selection analysis, it is important to improve the stability and efficiency of mRNA templates for both technologies. Here we describe an efficient single-strand ligation (90% of the input mRNAs) using a fluorescein-conjugated polyethylene glycol puromycin (Fluor-PEG Puro) spacer. This ligation provides a stable c-jun mRNA with a flexible Fluor-PEG Puro spacer for efficient fusion formation (70% of the input mRNA with the PEG spacer) in a cell-free wheat germ translation system. When using a 5' untranslated region including SP6 promoter and Omega29 enhancer (a part of tobacco mosaic virus Omega), an A(8) sequence (eight consecutive adenylate residues) at the 3' end is suitable for fusion formation, while an XA(8) sequence (XhoI and the A(8) sequence) is suitable for C-terminal protein labeling. Further, we report that Fluor-PEG N-t-butyloxycarbonylpuromycin [Puro(Boc)] spacer enhances the stability and efficiency of c-jun mRNA template for C-terminal protein labeling. These mRNA templates should be useful for puromycin-based technologies (fusion formation and C-terminal protein labeling) to facilitate high-throughput in vitro protein selection for not only evolutionary protein engineering, but also proteome exploration. PMID- 12888531 TI - Fast and accurate determination of sites along the FUT2 in vitro transcript that are accessible to antisense oligonucleotides by application of secondary structure predictions and RNase H in combination with MALDI-TOF mass spectrometry. AB - Alteration of gene expression by use of antisense oligonucleotides has considerable potential for therapeutic purposes and scientific studies. Although applied for almost 25 years, this technique is still associated with difficulties in finding antisense-effective regions along the target mRNA. This is mainly due to strong secondary structures preventing binding of antisense oligonucleotides and RNase H, playing a major role in antisense-mediated degradation of the mRNA. These difficulties make empirical testing of a large number of sequences complementary to various sites in the target mRNA a very lengthy and troublesome procedure. To overcome this problem, more recent strategies to find efficient antisense sites are based on secondary structure prediction and RNase H-dependent mechanisms. We were the first who directly combined these two strategies; antisense oligonucleotides complementary to predicted unpaired target mRNA regions were designed and hybridized to the corresponding RNAs. Incubation with RNase H led to cleavage of the RNA at the respective hybridization sites. Analysis of the RNA fragments by matrix-assisted laser desorption/ionization time of-flight (MALDI-TOF) mass spectrometry, which has not been used in this context before, allowed exact determination of the cleavage site. Thus the technique described here is very promising when searching for effective antisense sites. PMID- 12888532 TI - A simple two-step, 'hit and fix' method to generate subtle mutations in BACs using short denatured PCR fragments. AB - The bacteriophage lambda recombination system has proven to be a valuable tool for engineering bacterial artificial chromosomes (BAC). Due to its high efficiency, subtle alterations in the BACs can be generated using oligonucleotides as targeting vectors. Since no selection marker is used, recombinant clones are identified utilizing a selective PCR screening method. However, occasionally the selective PCR screening is not feasible. We describe here a two-step 'hit and fix' method that can be reliably used for generating any subtle alteration in BACs using short denatured PCR fragments as targeting vectors. In the first step of this method, 6-20 nucleotides are changed around the base where the mutation has to be generated. In the second step, these altered nucleotides are reverted to the original sequence and simultaneously a subtle alteration is introduced. Since in each step several nucleotides are changed, PCR primers specific for such alterations can be designed. This two-step method provides a simple and efficient tool for generating subtle alterations in BACs that can be very valuable for functional analysis of genes. PMID- 12888533 TI - Combination of overlapping bacterial artificial chromosomes by a two-step recombinogenic engineering method. AB - Recombinogenic engineering or recombineering is a powerful new method to engineer DNA without the need for restriction enzymes or ligases. We report here a general method for using recombineering to combine overlapping bacterial artificial chromosomes (BACs) to build larger, unified BACs. In order to test the feasibility of using recombineering to combine two large DNA fragments (>20 kb), we constructed a unified BAC containing the full-length tyrosinase-related protein-1 (Tyrp-1) gene from two library-derived BACs, one containing the 5' regulatory elements and the other containing the 3' coding exons. This was achieved using a two-step homologous recombination method enabled by the bacteriophage lambda Red proteins. In the first step, retrieval, a large DNA fragment (approximately 22 kb) was retrieved from one of the original BACs. In the second step, recombination, the retrieved DNA fragment was inserted into the second original BAC to form the unified BAC containing all the desired Tyrp-1 sequence. To further demonstrate the general applicability of our approach, an additional DNA fragment (approximately 20 kb) was inserted into the unified BAC downstream of the coding region. This method should prove very useful for enabling BAC manipulation in a variety of scenarios. PMID- 12888534 TI - General plasmids for producing RNA in vitro transcripts with homogeneous ends. AB - In vitro transcripts of bacteriophage RNA polymerases (RNAPs), such as T7 RNAP, often suffer from a considerable degree of 3'-end heterogeneity and, with certain promoter sequences, 5'-end heterogeneity. For some applications, this transcript heterogeneity poses a significant problem. A potential solution is to incorporate ribozymes into the transcripts at the 5'- and/or 3'-end of the target RNA sequence. This approach has been used quite widely but has required the generation of new transcription vectors or PCR-derived templates for each new RNA to be studied. To overcome this limitation, we have created two general plasmids for producing homogeneous RNA transcripts: one encodes a 3'- hepatitis delta virus (HDV) ribozyme and the other, used in combination with a two-step PCR, allows the production of double [5'-hammerhead (HH) and 3'-HDV] ribozyme constructs. A choice of cloning and run-off transcription linearisation restriction enzyme sites ensures that virtually any RNA sequence can be cloned and transcribed from these plasmids. For all the RNA sequences tested, good yields of transcript were obtained. These plasmids provide the tools for the simple, rapid creation of new RNA-coding plasmids to produce milligram quantities of homogeneous in vitro transcripts for all applications. PMID- 12888535 TI - Development of a rapid, small-scale DNA repair assay for use on clinical samples. AB - Double-strand breaks (DSBs) are the most lethal form of DNA damage. They can be repaired by one of two pathways, homologous recombination and non-homologous end joining (NHEJ). A NHEJ assay has previously been reported which measures joining using cell-free extracts and a linearised plasmid as DNA substrate. This assay was designed for 3 x 10(9) cells grown in vitro and utilised radioactively labelled substrate. We have scaled down the method to use smaller cell numbers in a variety of cell lines. Altering the cellular extraction procedure decreased background DNA contamination. The cleaner preparations allowed us to use SYBR Green I staining to identify joined products, which was as sensitive as 32P-end labelled DNA. NHEJ was found in established tumour cell lines from different originating tissues, though actual levels and fidelity of repair differed. This method also allowed end joining to be assessed in clinical specimens (human blood, brain and bladder tumours) within 24 h of receiving samples. The application of this method will allow investigation of the role of DSB DNA repair pathways in human tumours. PMID- 12888536 TI - Parallel competition analysis of Saccharomyces cerevisiae strains differing by a single base using polymerase colonies. AB - We describe a strategy to analyze the impact of single nucleotide mutations on protein function. Our method utilizes a combination of yeast functional complementation, growth competition of mutant pools and polyacrylamide gel immobilized PCR. A system was constructed in which the yeast PGK1 gene was expressed from a plasmid-borne copy of the gene in a PGK1 deletion strain of Saccharomyces cerevisiae. Using this system, we demonstrated that the enrichment or depletion of PGK1 point mutants from a mixed culture was consistent with the expected results based on the isolated growth rates of the mutants. Enrichment or depletion of individual point mutants was shown to result from increases or decreases, respectively, in the specific activities of the encoded proteins. Further, we demonstrate the ability to analyze the functional effect of many individual point mutations in parallel. By functional complementation of yeast deletions with human homologs, our technique could be readily applied to the functional analysis of single nucleotide polymorphisms in human genes of medical interest. PMID- 12888537 TI - Stable and heritable gene silencing in the malaria vector Anopheles stephensi. AB - Heritable RNA interference (RNAi), triggered from stably expressed transgenes with an inverted repeat (IR) configuration, is an important tool for reverse genetic studies. Here we report on the development of stable RNAi in Anopheles stephensi mosquitoes, the major vector of human malaria in Asia. Trans genic mosquitoes stably expressing a RNAi transgene, designed to produce intron-spliced double-stranded RNA (dsRNA) targeting the green fluorescent protein EGFP gene, were crossed to an EGFP-expressing target line. EGFP expression was dramatically reduced at both the protein and RNA levels. The levels of gene silencing depended upon the RNAi gene copy number and its site of integration. These results demonstrate that specific RNAi-mediated knockdown of gene function can be achieved with high efficiency in Anopheles . This will be invaluable to systematically unravel the function of Anopheles genes determining the vectorial capacity of the malaria parasite. PMID- 12888538 TI - Induction of cre recombinase activity using modified androgen receptor ligand binding domains: a sensitive assay for ligand-receptor interactions. AB - Novel systems of inducible gene expression are presented in which CRE-M, an altered form of cre recombinase (cre), is fused to and activated by ligand binding to two forms of the androgen receptor (AR) ligand binding domain (LBD). Selective activation or inactivation of gene transcription is induced upon the addition of appropriate ligand. The coupling of this cre-LBD system with our previously reported highly sensitive assay to measure cre activity in vitro using a dual fluorescent gene switch reporter provides a novel, high-throughput assay system for identifying compounds that bind to and activate various forms of the LBD of androgen receptor. This method can similarly be applied to screen compounds for their activating properties on other steroid hormone LBDs. Three different forms of the AR-LBD were fused to CRE-M, including the wild-type AR-LBD (wt), a non-ligand binding truncated form, LBD (T), and a mutated form (Thr-->Ala substitution) identified in the LNCaP prostate cancer cell line, LBD (LNCaP). We demonstrate a 10-fold induction of cre activity by the addition of androgen agonists to the CRE-M-AR-LBD(wt) fusion protein, but not in the presence of the anti-androgen, flutamide. However, cre activity can be induced by flutamide with the CRE-M-AR-LBD(LNCaP) fusion protein. Similar activation properties were obtained when these fusion proteins were expressed using adenoviral vectors. When combined with our previously reported cre-lox gene switch system, the CRE-M-AR LBD system can be utilized in gene therapy systems in which a therapeutic product may be initially expressed, replaced by a second product, or turned-off following exposure to ligand. This provides an important, additional level of regulation to gene therapy systems. PMID- 12888539 TI - Festina lente: evidences for fast and slow learning processes and a role for sleep in human motor skill learning. PMID- 12888540 TI - Remembering people: neuroimaging takes on the real world. PMID- 12888541 TI - Diencephalic damage decreases hippocampal acetylcholine release during spontaneous alternation testing. AB - A rodent model of diencephalic amnesia, pyrithiamine-induced thiamine deficiency (PTD), was used to investigate diencephalic-hippocampal interactions. Acetylcholine (ACh) release, a marker of memory-related activation, was measured in the hippocampus of PTD-treated and control rats prior to, during, and after spontaneous alternation test. During behavioral testing, all animals displayed increases in ACh release. However, both the percent increase of ACh release during spontaneous alternation testing and the alternation scores were higher in control rats relative to PTD-treated rats. Thus, when rats are tested on a task with demands dependent on the hippocampus, it appears that the hippocampus is not fully activated after diencephalic damage. PMID- 12888542 TI - Molecular mechanisms contributing to long-lasting synaptic plasticity at the temporoammonic-CA1 synapse. AB - The hippocampus and the nearby medial temporal lobe structures are required for the formation, consolidation, and retrieval of episodic memories. Sensory information enters the hippocampus via two inputs from entorhinal cortex (EC): One input (perforant path) makes synapses on the dendrites of dentate granule cells as the first set of synapses in the trisynaptic circuit, the other (temporoammonic; TA) makes synapses on the distal dendrites of CA1 neurons. Here we demonstrate that TA-CA1 synapses undergo both early- and late-phase long-term potentiation (LTP) in rat hippocampal slices. LTP at TA-CA1 synapses requires both NMDA receptor and voltage-gated Ca2+ channel activity. Furthermore, TA-CA1 LTP is insensitive to the blockade of fast inhibitory transmission (GABAA mediated) and, interestingly, is dependent on GABAB-dependent slow inhibitory transmission. These findings indicate that the TA-CA1 synapses may rely on a refined modulation of inhibition to exhibit LTP. PMID- 12888543 TI - Neural correlates of person recognition. AB - Rapidly identifying known individuals is an essential skill in human society. To elucidate the neural basis of this skill, we monitored brain activity while experimental participants demonstrated their ability to recognize people on the basis of viewing their faces. Each participant first memorized the faces of 20 individuals who were not known to the participants in advance. Each face was presented along with a voice simulating the individual speaking their name and a biographical fact. Following this learning procedure, the associated verbal information could be recalled accurately in response to each face. These learned faces were subsequently viewed together with new faces in a memory task. Subjects made a yes-no recognition decision in response to each face while also covertly retrieving the person-specific information associated with each learned face. Brain activity that accompanied this retrieval of person-specific information was contrasted to that when new faces were processed. Functional magnetic resonance imaging in 10 participants showed that several brain regions were activated during blocks of learned faces, including left hippocampus, left middle temporal gyrus, left insula, and bilateral cerebellum. Recordings of event-related brain potentials in 10 other participants tracked the time course of face processing and showed that learned faces engaged neural activity responsible for person recognition 300-600 msec after face onset. Collectively, these results suggest that the visual input of a recently learned face can rapidly trigger retrieval of associated person-specific information through reactivation of distributed cortical networks linked via hippocampal connections. PMID- 12888545 TI - Enhanced human memory consolidation with post-learning stress: interaction with the degree of arousal at encoding. AB - Abundant evidence indicates that endogenous stress hormones such as epinephrine and corticosterone modulate memory consolidation in animals. We recently provided the first demonstration that an endogenous stress hormone (epinephrine) can enhance human memory consolidation. However, these findings also suggested that post-learning stress hormone activation does not uniformly enhance memory for all recently acquired information; rather, that it interacts with the degree of arousal at initial encoding of material in modulating memory for the material. Here we tested this hypothesis by administering cold pressor stress (CPS) or a control procedure to subjects after they viewed slides of varying emotional content, and assessing memory for the slides 1 wk later. CPS, which significantly elevated salivary cortisol levels, enhanced memory for emotionally arousing slides compared with the controls, but did not affect memory for relatively neutral slides. These findings further support the view that post-learning stress hormone-related activity interacts with arousal at initial encoding to modulate memory consolidation. PMID- 12888544 TI - Spared anterograde memory for shock-probe fear conditioning after inactivation of the amygdala. AB - Previous studies have shown that amygdala lesions impair avoidance of an electrified probe. This finding has been interpreted as indicating that amygdala lesions reduce fear. It is unclear, however, whether amygdala-lesioned rats learn that the probe is associated with shock. If the lesions prevent the formation of this association, then pretraining reversible inactivation of the amygdala should impair both acquisition and retention performance. To test this hypothesis, the amygdala was inactivated (tetrodotoxin; TTX; 1 ng/side) before a shock-probe acquisition session, and retention was tested 4 d later. The data indicated that, compared with rats infused with vehicle, rats infused with TTX received more shocks during the acquisition session, but more importantly, were not impaired on the retention test. In Experiment 2, we assessed whether the spared memory on the retention test was caused by overtraining during acquisition. We used the same procedure as in Experiment 1, with the exception that the number of shocks the rats received during the acquisition session was limited to four. Again the data indicated that amygdala inactivation did not impair performance on the retention test. These results indicate that amygdala inactivation does not prevent the formation of an association between the shock and the probe and that shock-probe deficits during acquisition likely reflect the amygdala's involvement in other processes. PMID- 12888546 TI - Sleep and the time course of motor skill learning. AB - Growing evidence suggests that sleep plays an important role in the process of procedural learning. Most recently, sleep has been implicated in the continued development of motor-skill learning following initial acquisition. However, the temporal evolution of motor learning before and after sleep, the effects of different training regimens, and the long-term development of motor learning across multiple nights of sleep remain unknown. Here, we report data for subjects trained and retested on a sequential finger-tapping task across multiple days. The findings demonstrate firstly that following initial training, small practice dependent improvements are possible before, but not following the large practice independent gains that develop across a night of sleep. Secondly, doubling the quantity of initial training does not alter the amount of subsequent sleep dependent learning that develops overnight. Thirdly, the amount of sleep dependent learning does not correlate with the amount of practice-dependent learning achieved during training, suggesting the existence of two discrete motor learning processes. Finally, whereas the majority of sleep-dependent motor-skill learning develops during the first night of sleep following training, additional nights of sleep still offer continued improvements. PMID- 12888547 TI - Differential involvement of NMDA and AMPA receptors within the nucleus accumbens in consolidation of information necessary for place navigation and guidance strategy of mice. AB - Recent evidence now points to a role of glutamate transmission within the nucleus accumbens (Nacc) in spatial learning and memory. Unfortunately, the role of the distinct classes of glutamate receptors within this structure in mediating the different steps of the memorization process is not clear. The aim of this study therefore was to further investigate this issue, trying to assess the involvement of the two classes of glutamate receptors within the Nacc in consolidation of spatial information using an associative spatial task, the water maze. For this purpose, focal injections of the NMDA antagonist, AP-5, and of the AMPA antagonist, DNQX, have been performed immediately after the training phase, and mice have been tested for retention 24 h later. Two different versions of the water-maze task have been used: In the place version, animals could learn the position of the platform using visual distal cues, and in the cue version, the location of the platform was indicated by a single proximal cue. The results demonstrated that posttraining NMDA receptor blockade affects mice response in the place but not in the cue water-maze task. On the contrary, AMPA receptor blockade induced no effect in either version of the task. These data confirm a functional dissociation between glutamate receptors located in the Nacc in modulating spatial memory consolidation and indicate that they are specifically involved in consolidation of information necessary to acquire a place but not to a guidance strategy. PMID- 12888548 TI - Complex effects of NMDA receptor antagonist APV in the basolateral amygdala on acquisition of two-way avoidance reaction and long-term fear memory. AB - Although much has been learned about the role of the amygdala in Pavlovian fear conditioning, relatively little is known about an involvement of this structure in more complex aversive learning, such as acquisition of an active avoidance reaction. In the present study, rats with a pretraining injection of the N-methyl D-aspartate (NMDA) receptor antagonist, 2-amino-5-phosphonopentanoic acid (APV), into the basolateral amygdala (BLA) were found to be impaired in two-way active avoidance learning. During multitrial training in a shuttle box, the APV-injected rats were not different from the controls in sensitivity to shock or in acquisition of freezing to contextual cues. However, APV injection led to impaired retention of contextual fear when tested 48 h later, along with an attenuation of c-Fos expression in the amygdala. These results are consistent with the role of NMDA receptors of the BLA in long-term memory of fear, previously documented in Pavlovian conditioning paradigms. The APV-induced impairment in the active avoidance learning coincided with deficits in directionality of the escape reaction and in attention to conditioned stimuli. These data indicate that normal functioning of NMDA receptors in the basolateral amygdala is required during acquisition of adaptive instrumental responses in a shuttle box but is not necessary for acquisition of short-term contextual fear in this situation. PMID- 12888549 TI - Astrocyte protection of neurons: role of transforming growth factor-beta signaling via a c-Jun-AP-1 protective pathway. AB - Astrocytes have become a focal point for research in neurobiology, especially regarding their purported ability to regulate neuronal communication and survival. The present study addressed a poorly understood but important focus in this area, the mechanism(s) underlying astrocyte-induced survival of neurons. The results of the study show that soluble factors in astrocyte-conditioned media (ACM) protect murine GT1-7 neurons from serum deprivation-induced cell death and that this neuroprotection is correlated with enhanced activation/phosphorylation of the AP-1 transcription factor, c-JunSer-63. A parallel and correlated activation of the upstream kinases, c-Jun N-terminal kinase (JNK) and mitogen activated protein kinase kinase-4 (MKK4) was also demonstrated. Furthermore, co administration of JNK inhibitors, but not a MEK inhibitor, significantly attenuated ACM-induced phosphorylation of c-JunSer-63 and blocked its neuroprotective action. Gel shift analysis demonstrated that ACM enhanced AP-1 binding, an effect that appears functionally important, since an AP-1 binding inhibitor significantly attenuated the neuroprotective action of ACM. Further studies implicated transforming growth factor (TGF)-beta1 and TGF-beta2 as critical active soluble factors released by astrocytes, since both were demonstrated in ACM, and immunoneutralization of the conditioned media with a panspecific TGF-beta antibody significantly attenuated the enhanced AP-1 binding and neuroprotective action of the ACM. Furthermore, exogenous application of TGF beta1 and TGF-beta2 was found to enhance c-JunSer-63 phosphorylation and to be neuroprotective, and co-administration of JNK inhibitors or an AP-1 binding inhibitor blocked TGF-beta-induced neuroprotection. Taken together, these studies suggest that astrocytes can protect neurons from serum deprivation-induced cell death, at least in part, by release of TGF-beta and activation of a c-Jun/AP-1 protective pathway. PMID- 12888550 TI - Oligomerization of the alpha 1a- and alpha 1b-adrenergic receptor subtypes. Potential implications in receptor internalization. AB - We combined biophysical, biochemical, and pharmacological approaches to investigate the ability of the alpha 1a- and alpha 1b-adrenergic receptor (AR) subtypes to form homo- and hetero-oligomers. Receptors tagged with different epitopes (hemagglutinin and Myc) or fluorescent proteins (cyan and green fluorescent proteins) were transiently expressed in HEK-293 cells either individually or in different combinations. Fluorescence resonance energy transfer measurements provided evidence that both the alpha 1a- and alpha 1b-AR can form homo-oligomers with similar transfer efficiency of approximately 0.10. Hetero oligomers could also be observed between the alpha 1b- and the alpha 1a-AR subtypes but not between the alpha 1b-AR and the beta2-AR, the NK1 tachykinin, or the CCR5 chemokine receptors. Oligomerization of the alpha 1b-AR did not require the integrity of its C-tail, of two glycophorin motifs, or of the N-linked glycosylation sites at its N terminus. In contrast, helix I and, to a lesser extent, helix VII were found to play a role in the alpha 1b-AR homo oligomerization. Receptor oligomerization was not influenced by the agonist epinephrine or by the inverse agonist prazosin. A constitutively active (A293E) as well as a signaling-deficient (R143E) mutant displayed oligomerization features similar to those of the wild type alpha 1b-AR. Confocal imaging revealed that oligomerization of the alpha1-AR subtypes correlated with their ability to co-internalize upon exposure to the agonist. The alpha 1a-selective agonist oxymetazoline induced the co-internalization of the alpha 1a- and alpha 1b-AR, whereas the alpha 1b-AR could not co-internalize with the NK1 tachykinin or CCR5 chemokine receptors. Oligomerization might therefore represent an additional mechanism regulating the physiological responses mediated by the alpha 1a- and alpha 1b-AR subtypes. PMID- 12888551 TI - Ligand-mediated activation of the cAMP-responsive guanine nucleotide exchange factor Epac. AB - Epac is a cAMP-dependent exchange factor for the small GTP-binding protein Rap. The activity of Epac is inhibited by a direct interaction between the C-terminal helical part of the cAMP-binding domain, called the lid, and the catalytic region, which is released after binding of cAMP. Herein, we show that the activation properties are very sensitive to modifications of the cyclic nucleotide. Some analogues are inhibitory and others are stimulatory; some are characterized by a much higher activation potential than normal cAMP. Mutational analysis of Epac allows insights into a network of interactions between the cyclic nucleotides and Epac. Mutations in the lid region are able to amplify or to attenuate selectively the activation potency of cAMP analogues. The properties of cAMP analogues previously used for the activation of the cAMP responsive protein kinase A and of 8-(4-chlorophenylthio)-2'-O-methyladenosine-3',5' cyclicmonophosphate, an analogue highly selective for activation of Epac were investigated in detail. PMID- 12888552 TI - Gene knockdown of gamma-glutamylcysteine synthetase by RNAi in the parasitic protozoa Trypanosoma brucei demonstrates that it is an essential enzyme. AB - The parasitic protozoa Trypanosoma brucei utilizes a novel cofactor (trypanothione, T(SH)2), which is a conjugate of GSH and spermidine, to maintain cellular redox balance. gamma-Glutamylcysteine synthetase (gamma-GCS) catalyzes the first step in the biosynthesis of GSH. To evaluate the importance of thiol metabolism to the parasite, RNAi methods were used to knock down gene expression of gamma-GCS in procyclic T. brucei cells. Induction of gamma-GCS RNAi with tetracycline led to cell death within 4-6 days post-induction. Cell death was preceded by the depletion of the gamma-GCS protein and RNA and by the loss of the cellular pools of GSH and T(SH)2. The addition of GSH (80 microM) to cell cultures rescued the RNAi cell death phenotype and restored the intracellular thiol pools to wild-type levels. Treatment of cells with buthionine sulfoximine (BSO), an enzyme-activated inhibitor of gamma-GCS, also resulted in cell death. However, the toxicity of the inhibitor was not reversed by GSH, suggesting that BSO has more than one cellular target. BSO depletes intracellular thiols to a similar extent as gamma-GCS RNAi; however, addition of GSH did not restore the pools of GSH and T(SH)2. These data suggest that BSO also acts to inhibit the transport of GSH or its peptide metabolites into the cell. The ability of BSO to inhibit both synthesis and transport of GSH likely makes it a more effective cytotoxic agent than an inhibitor with a single mode of action. Finally the potential for the T(SH)2 biosynthetic enzymes to be regulated in response to reduced thiol levels was studied. The expression levels of ornithine decarboxylase and of S-adenosylmethionine decarboxylase, two essential enzymes in spermidine biosynthesis, remained constant in induced gamma-GCS RNAi cell lines. PMID- 12888553 TI - The intracellular domain of the low density lipoprotein receptor-related protein modulates transactivation mediated by amyloid precursor protein and Fe65. AB - Low density lipoprotein-related protein (LRP) is a transmembrane receptor, localized mainly in hepatocytes, fibroblasts, and neurons. It is implicated in diverse biological processes both as an endocytic receptor and as a signaling molecule. Recent reports show that LRP undergoes sequential proteolytic cleavage in the ectodomain and transmembrane domain. The latter cleavage, mediated by the Alzheimer-related gamma-secretase activity that also cleaves amyloid precursor protein (APP) and Notch, results in the release of the LRP cytoplasmic domain (LRPICD) fragment. This relatively small cytoplasmic fragment has several motifs by which LRP interacts with various intracellular adaptor and scaffold proteins. However, the function of this fragment is largely unknown. Here we show that the LRPICD is translocated to the nucleus, where it colocalizes in the nucleus with a transcription modulator, Tip60, which is known to interact with Fe65 and with the APP-derived intracellular domain. LRPICD dramatically inhibits APP-derived intracellular domain/Fe65 transactivation mediated by Tip60. LRPICD has a close interaction with Tip60 in the nucleus, as shown by a fluorescence resonance energy transfer assay. These observations suggest that LRPICD has a novel signaling function, negatively impacting transcriptional activity of the APP, Fe65, and Tip60 complex in the nucleus, and shed new light on the function of LRP in transcriptional modulation. PMID- 12888554 TI - HAMLET interacts with histones and chromatin in tumor cell nuclei. AB - HAMLET is a folding variant of human alpha-lactalbumin in an active complex with oleic acid. HAMLET selectively enters tumor cells, accumulates in their nuclei and induces apoptosis-like cell death. This study examined the interactions of HAMLET with nuclear constituents and identified histones as targets. HAMLET was found to bind histone H3 strongly and to lesser extent histones H4 and H2B. The specificity of these interactions was confirmed using BIAcore technology and chromatin assembly assays. In vivo in tumor cells, HAMLET co-localized with histones and perturbed the chromatin structure; HAMLET was found associated with chromatin in an insoluble nuclear fraction resistant to salt extraction. In vitro, HAMLET bound strongly to histones and impaired their deposition on DNA. We conclude that HAMLET interacts with histones and chromatin in tumor cell nuclei and propose that this interaction locks the cells into the death pathway by irreversibly disrupting chromatin organization. PMID- 12888555 TI - The metalloproteolytic activity of the anthrax lethal factor is substrate inhibited. AB - The anthrax lethal factor (LF) is a Zn2+ endopeptidase specific for mitogen activated protein kinase kinases (MAPKKs), which are cleaved within their N termini. Here, the proteolytic activity of LF has been investigated using novel chromogenic MAPKK-derived peptide substrates, which allowed us to determine the kinetic parameters of the reaction. LF displayed maximal proteolytic activity at the pH and temperature values of the cell cytosol, which is its site of action. LF undergoes substrate inhibition, in keeping with the non-productive binding geometry of the MAPPK-2 N terminus to LF. PMID- 12888556 TI - Crystal structure of OxyC, a cytochrome P450 implicated in an oxidative C-C coupling reaction during vancomycin biosynthesis. AB - Gene inactivation studies point to the involvement of OxyC in catalyzing the last oxidative phenol coupling reaction during glycopeptide antibiotic biosynthesis. Presently, the substrate and exact timing of the OxyC reaction are unknown. The substrate might be the bicyclic heptapeptide or a thioester derivative bound to a protein carrier domain. OxyC from the vancomycin producer Amycolatopsis orientalis was produced in Escherichia coli and crystallized, and its structure was determined to 1.9 A resolution. OxyC gave UV-visible spectra characteristic of a P450-like hemoprotein in the low spin ferric state. After reduction to the ferrous state by dithionite the CO-ligated form gave a 450-nm peak in a UV difference spectrum. The addition of vancomycin aglycone to OxyC produced type I changes to the UV spectrum. OxyC exhibits the typical P450-fold, with the Cys ligand loop containing the signature sequence FGHGX-HXCLG and Cys-356 being the proximal axial thiolate ligand of the heme iron. The observation of a water molecule bound to the heme iron is consistent with the UV-visible spectra of OxyC indicating a low spin heme. A polyethylene glycol molecule occupying the active site might mimic the bicyclic heptapeptide substrate. Analysis of the structure of Oxy-proteins and other P450s indicates regions that might be involved in binding of the redox partner and possibly the protein carrier domain. PMID- 12888557 TI - The binding site for regulatory 14-3-3 protein in plant plasma membrane H+ ATPase: involvement of a region promoting phosphorylation-independent interaction in addition to the phosphorylation-dependent C-terminal end. AB - 14-3-3 proteins constitute a family of well conserved proteins interacting with a large number of phosphorylated binding partners in eukaryotic cells. The plant plasma membrane H+-ATPase is an unusual target in that a unique phosphothreonine motif (946YpTV, where pT represents phosphothreonine) in the extreme C-terminal end of the H+-ATPase interacts with the binding cleft of 14-3-3 protein (Wurtele, M., Jelich-Ottmann, C., Wittinghofer, A., and Oecking, C. (2003) EMBO J. 22, 987 994). We report binding of 14-3-3 protein to a nonphosphorylated peptide representing the 34 C-terminal residues of the Arabidopsis plasma membrane H+ ATPase isoform 2 (AHA2). Following site-directed mutagenesis within the 45 C terminal residues of AHA2, we conclude that, in addition to the 946YpTV motif, a number of residues located further upstream are required for phosphorylation independent binding of 14-3-3. Among these, Thr-924 is important for interaction with 14-3-3 protein even when Thr-947 is phosphorylated. We suggest that the role of phosphorylation, which is accentuated by fusicoccin, is to stabilize protein protein interaction between 14-3-3 protein and several residues of the H+-ATPase C-terminal domain. PMID- 12888558 TI - Ligand-independent CXCR2 dimerization. AB - Homo- and hetero-oligomerization have been reported for several G protein-coupled receptors (GPCRs). The CXCR2 is a GPCR that is activated, among the others, by the chemokines CXCL8 (interleukin-8) and CXCL2 (growth-related gene product beta) to induce cell chemotaxis. We have investigated the oligomerization of CXCR2 receptors expressed in human embryonic kidney cells and generated a series of truncated mutants to determine whether they could negatively regulate the wild type (wt) receptor functions. CXCR2 receptor oligomerization was also studied by coimmunoprecipitation of green fluorescent protein- and V5-tagged CXCR2. Truncated CXCR2 receptors retained their ability to form oligomers only if the region between the amino acids Ala-106 and Lys-163 was present. In contrast, all of the deletion mutants analyzed were able to form heterodimers with the wt CXCR2 receptor, albeit with different efficiency, competing for wt/wt dimer formation. The truncated CXCR2 mutants were not functional and, when coexpressed with wt CXCR2, interfered with receptor functions, impairing cell signaling and chemotaxis. When CXCR2 was expressed with the AMPA-type glutamate receptor GluR1, CXCR2 dimerization was again impaired in a dose-dependent way, and receptor functions were prejudiced. In contrast, CXCR1, a chemokine receptor that shares many similarities with CXCR2, did not dimerize alone or with CXCR2 and when coexpressed with CXCR2 did not impair receptor signaling and chemotaxis. The formation of CXCR2 dimers was also confirmed in cerebellar neuron cells. Taken together, we conclude from these studies that CXCR2 functions as a dimer and that truncated receptors negatively modulate receptor activities competing for the formation of wt/wt dimers. PMID- 12888559 TI - Concomitant increase of histone acetyltransferase activity and degradation of p300 during retinoic acid-induced differentiation of F9 cells. AB - The p300 and closely related CBP histone acetyltransferases (HAT) function as global transcriptional co-activators that play roles in many cell differentiation and signal transduction pathways. Despite their similarities, p300 and CBP have distinct functions during retinoic acid-induced differentiation of mouse F9 embryonal carcinoma cells. F9 cells constitute a well established model system for investigating the first steps of early development and retinoic acid signaling ex vivo. p300, but not CBP, was shown to be essential for F9 differentiation. In this study we have investigated the regulation of p300 during F9 differentiation. We report a dramatic decrease of p300, but not CBP protein levels, after 48 h of retinoic acid treatment. p300 is degraded via the ubiquitin proteasome pathway. Although the large majority of p300 is degraded, its global HAT activity stays constant during F9 differentiation, which means that its specific HAT activity increases considerably. p300 is strongly phosphorylated in both undifferentiated and differentiated F9 cells; its HAT activity, however, is independent of phosphorylation before differentiation and becomes dependent on phosphorylation during differentiation. Furthermore, we show that protein kinase A affects p300 HAT activity both in vivo and in vitro as well as p300 phosphorylation in differentiated cells. Thus, we show that p300 is differentially phosphorylated in undifferentiated versus differentiated cells and that the changes in phosphorylation affect its HAT activity. Moreover, our study suggests an explanation for the functional switch of p300-mediated repression versus activation during F9 differentiation. PMID- 12888560 TI - Aurintricarboxylic acid blocks in vitro and in vivo activity of YopH, an essential virulent factor of Yersinia pestis, the agent of plague. AB - Yersinia are causative agents in human diseases ranging from gastrointestinal syndromes to Bubonic Plague. There is increasing risk of misuse of infectious agents, such as Yersinia pestis, as weapons of terror as well as instruments of warfare for mass destruction. YopH is an essential virulence factor whose protein tyrosine phosphatase (PTP) activity is required for Yersinia pathogenicity. Consequently, there is considerable interest in developing potent and selective YopH inhibitors as novel anti-plague agents. We have screened a library of 720 structurally diverse commercially available carboxylic acids and identified 26 YopH inhibitors with IC50 values below 100 mum. The most potent and specific YopH inhibitor is aurintricarboxylic acid (ATA), which exhibits a Ki value of 5 nm for YopH and displays 6-120-fold selectivity in favor of YopH against a panel of mammalian PTPs. To determine whether ATA can block the activity of YopH in a cellular context, we have examined the effect of ATA on T-cell signaling in human Jurkat cells transfected with YopH. We show that YopH severely decreases the T cell receptor-induced cellular tyrosine phosphorylation, ERK1/2 activity, and interleukin-2 transcriptional activity. We demonstrate that ATA can effectively block the inhibitory activity of YopH and restore normal T-cell function. These results provide a proof-of-concept for the hypothesis that small molecule inhibitors that selectively target YopH may be therapeutically useful. In addition, it is expected that potent and selective YopH inhibitors, such as ATA, should be useful reagents to delineate YopH's cellular targets in plague and other pathogenic conditions caused by Yersinia infection. PMID- 12888561 TI - Myeloperoxidase-derived hypochlorous acid antagonizes the oxidative stress mediated activation of iron regulatory protein 1. AB - Hypochlorous acid (HOCl) is a highly reactive product generated by the myeloperoxidase reaction during the oxidative burst of activated neutrophils, which is implicated in many bactericidal and cytotoxic responses. Recent evidence suggests that HOCl may also play a role in the modulation of redox sensitive signaling pathways. The short half-life of HOCl and the requirement for a continuous presence of H2O2 as a substrate for its myeloperoxidase-catalyzed generation make the study of HOCl-mediated responses very difficult. We describe here an enzymatic model consisting of glucose/glucose oxidase, catalase, and myeloperoxidase (GOX/CAT/MPO) that allows the controlled generation of both HOCl and H2O2 and thus, mimics the oxidative burst of activated neutrophils. By employing this model we show that HOCl prevents the H2O2-mediated activation of iron regulatory protein 1 (IRP1), a central post-transcriptional regulator of mammalian iron metabolism. Activated IRP1 binds to (R)iron-responsive elements" (IREs) within the mRNAs encoding proteins of iron metabolism and thereby controls their translation or stability. The inhibitory effect of HOCl is not a result of a direct modification of IRP1 by this oxidant. Kinetics experiments provide evidence that HOCl intervenes with the signaling cascade, which results in the activation of IRP1. We further demonstrate that HOCl antagonizes the H2O2 mediated increase in the levels of transferrin receptor, which is a downstream target of IRP1. Our findings suggest that HOCl can modulate signaling pathways in a concerted action with H2O2. The GOX/CAT/MPO system provides a valuable tool for studying the regulatory function of HOCl. PMID- 12888562 TI - Requirement of Cys399 for processing of the human ecto-ATPase (NTPDase2) and its implications for determination of the activities of splice variants of the enzyme. AB - Ecto-ATPase (CD39L1) corresponds to the type 2 enzyme of the ecto-nucleoside triphosphate diphosphohydrolase family (E-NTPDase). We have isolated from human ECV304 cells three cDNAs with high homology with members of the E-NTPDase family that encode predicted proteins of 495, 472, and 450 amino acids. Sequencing of a genomic DNA clone confirmed that these three sequences correspond to splice variants of the human ecto-ATPase (NTPDase2 alpha,-2 beta, and -2 gamma). Although all three enzyme forms were expressed heterologously to similar levels in Chinese hamster ovary cells clone K-1 (CHO-K1) cells, only the 495-amino acid protein (NTPDase2 alpha exhibited ecto-ATPase activity. Immunolocalization studies demonstrated that NTPDase2 alpha is fully processed and trafficked to the plasma membrane, whereas the NTPDase2 beta and -2 gamma splice variants were retained in not fully glycosylated forms in the endoplasmic reticulum. The potential roles of two highly conserved residues, Cys399 and Asn443, in the activity and cellular trafficking of the ecto-ATPase were examined. Mutation of Cys399, which is absent in NTPDase2 beta and -2 gamma, produced a protein completely devoid of nucleotidase activity, while mutation of Asn443 to Asp resulted in substantial loss of activity. Neither the Cys399 nor Asn443 mutants were fully glycosylated, and both were retained in the endoplasmic reticulum. These results indicate that the lack of ecto-nucleotidase activity exhibited by NTPDase2 beta and -2 gamma and the C399S mutant, as well as the large reduction of activity in the N443D mutant are due to alterations in the folding/maturation of these proteins. PMID- 12888563 TI - A reassessment of the effects of luminal [Ca2+] on inositol 1,4,5-trisphosphate induced Ca2+ release from internal stores. AB - Inositol 1,4,5-trisphosphate (InsP3)-induced Ca2+ release from intracellular stores displays complex kinetic behavior. While it well established that cytosolic [Ca2+] can modulate release by acting on the InsP3 receptor directly, the role of the filling state of internal Ca2+stores in modulating Ca2+ release remains unclear. Here we have reevaluated this topic using a technique that permits rapid and reversible changes in free [Ca2+] in internal stores of living intact cells without altering cytoplasmic [Ca2+], InsP3 receptors, or sarcoendoplasmic reticulum Ca2+ ATPases (SERCAs). N,N,N',N'-Tetrakis(2 pyridylmethyl)ethylene diamine (TPEN), a membrane-permeant, low affinity Ca2+ chelator was used to manipulate [Ca2+] in intracellular stores, while [Ca2+] changes within the store were monitored directly with the low-affinity Ca2+ indicator, mag-fura-2, in intact BHK-21 cells. 200 microM TPEN caused a rapid drop in luminal free [Ca2+] and significantly reduced the extent of the response to stimulation with 100 nm bradykinin, a calcium-mobilizing agonist. The same effect was observed when intact cells were pretreated with 1,2-bis(2 aminophenoxy)ethane-N,N,N',N'-tetraacetic acid(acetoxymethyl ester) (BAPTA-AM) to buffer cytoplasmic [Ca2+] changes. Although inhibition of Ca2+ uptake using the SERCA inhibitor tBHQ permitted significantly larger release of Ca2+ from stores, TPEN still attenuated the release in the presence of tBHQ in BAPTA-AM-loaded cells. These results demonstrate that the filling state of stores modulates the magnitude of InsP3-induced Ca2+release by additional mechanism(s) that are independent of regulation by cytoplasmic [Ca2+] or effects on SERCA pumps. PMID- 12888564 TI - An evolutionarily conserved N-terminal acetyltransferase complex associated with neuronal development. AB - We previously identified mNAT1 (murine N-terminal acetyltransferase 1) as an embryonic gene that is expressed in the developing brain and subsequently down regulated, in part, by the onset of N-methyl-d-aspartate (NMDA) receptor function. By searching the data base we discovered a second closely related gene, mNAT2. mNAT1 and mNAT2 are highly homologous to yeast NAT1, a gene known to regulate entry into the G0 phase of the cell cycle. However, in the absence of further characterization, including evidence that mammalian homologues of NAT1 encode functional acetyltransferases, the significance of this relationship has been unclear. Here we focus on mNAT1. Biochemical analysis demonstrated that mNAT1 and its evolutionarily conserved co-subunit, mARD1, assemble to form a functional acetyltransferase. Transfection of mammalian cells with mNAT1 and mARD1 followed by immunofluorescent staining revealed that these proteins localize to the cytoplasm in both overlapping and separate compartments. In situ hybridization demonstrated that throughout brain development mNAT1 and mARD1 are highly expressed in areas of cell division and migration and are down-regulated as neurons differentiate. Finally, mNAT1 and mARD1 are expressed in proliferating mouse P19 embryonic carcinoma cells; treatment of these cells with retinoic acid initiates exit from the cell cycle, neuronal differentiation, and down-regulation of mNAT1 and mARD1 as the NMOA receptor 1 gene is induced. The results provide the first direct evidence that vertebrate homologues of NAT1 and ARD1 form an evolutionarily conserved N-terminal acetyltransferase and suggest that expression and down-regulation of this enzyme complex plays an important role in the generation and differentiation of neurons. PMID- 12888565 TI - The blood group P1 synthase gene is identical to the Gb3/CD77 synthase gene. A clue to the solution of the P1/P2/p puzzle. AB - Blood group P1/P2 is a glycolipid antigen system for which the genetic mechanism has not yet been clarified. We analyzed the potential of the cloned Gb3/CD77 synthase to synthesize P1 antigen, because Gb3/CD77 and P1 share a common structure, Galalpha1,4Galbeta1,4Glc (NAc)-. L cell transfectants with Gb3/CD77 synthase cDNA expressed marginal levels of P1 on the cell surface but contained high levels of P1 in the cytoplasm. P2-type erythrocytes, which were serotyped as P2, also contained definite P1 antigen inside cells, although the amounts were lower than those of P1 cells. Only p erythrocytes lacked P1 antigen corresponding with function-losing mutations in the Gb3/CD77 synthase gene. Synthesis of P1 antigen from paragloboside in vitro was demonstrated using membrane fraction of the transfectants and a fusion enzyme with protein A. These results strongly suggested that P1 synthase is identical to Gb3/CD77 synthase and appear to propose a clue for the solution of the long-pending P1/P2/p puzzle. The P1/P2 difference might result from the difference in P1 quantity based on either different enzyme activity or the presence/absence of other enzyme modulators. Because P2 erythrocytes showed lower levels of Gb3/CD77 synthase mRNA than P1, 5' upstream promoter regions were analyzed, resulting in the identification of two P2-specific homozygous mutations. Differences in the transcriptional regulation in erythrocytes might be a major factor determining P1/P2. PMID- 12888567 TI - Calcium signaling regulates translocation and activation of Rac. AB - Rac is activated in response to various stimuli including growth factors and by adhesion to the extracellular matrix. However, how these stimuli ultimately result in Rac activation is poorly understood. The increase in intracellular calcium [Ca2+]i represents a ubiquitous second messenger system in cells, linking receptor activation to downstream signaling pathways. Here we show that elevation of [Ca2+]i, either artificially or by thrombin receptor activation, potently induces Rac activation. Lamellipodia formation induced by artificial elevation of [Ca2+]i is blocked by inhibition of Rac signaling, indicating that calcium induced cytoskeletal changes are controlled by the activation of Rac. Calcium dependent Rac activation was dependent on the activation of a conventional protein kinase C. Furthermore, both increased [Ca2+]i and protein kinase C activation induce phosphorylation of RhoGDI alpha and induce the translocation of cytosolic Rac to the plasma membrane. Intracellular calcium signaling may thus contribute to the intracellular localization and activation of Rac to regulate the cytoskeletal changes in response to receptor stimulation. PMID- 12888566 TI - Structural complementarity of Toll/interleukin-1 receptor domains in Toll-like receptors and the adaptors Mal and MyD88. AB - The Toll/interleukin 1 receptor (TIR) domain is a region found in the cytoplasmic tails of members of the Toll-like receptor/interleukin-1 receptor superfamily. The domain is essential for signaling and is also found in the adaptor proteins Mal (MyD88 adaptor-like) and MyD88, which function to couple activation of the receptor to downstream signaling components. Experimental structures of two Toll/interleukin 1 receptor domains reveal a alpha-beta-fold similar to that of the bacterial chemotaxis protein CheY, and other evidence suggests that the adaptors can make heterotypic interactions with both the receptors and themselves. Here we show that the purified TIR domains of Mal and MyD88 can form stable heterodimers and also that Mal homodimers and oligomers are dissociated in the presence of ATP. To identify structural features that may contribute to the formation of signaling complexes, we produced models of the TIR domains from human Toll-like receptor 4 (TLR4), Mal, and MyD88. We found that although the overall fold is conserved the electrostatic surface potentials are quite distinct. Docking studies of the models suggest that Mal and MyD88 bind to different regions in TLRs 2 and 4, a finding consistent with a cooperative role of the two adaptors in signaling. Mal and MyD88 are predicted to interact at a third non-overlapping site, suggesting that the receptor and adaptors may form heterotetrameric complexes. The theoretical model of the interactions is supported by experimental data from glutathione S-transferase pull-downs and co immunoprecipitations. Neither theoretical nor experimental data suggest a direct role for the conserved proline in the BB-loop in the association of TLR4, Mal, and MyD88. Finally we show a sequence relationship between the Drosophila protein Tube and Mal that may indicate a functional equivalence of these two adaptors in the Drosophila and vertebrate Toll pathways. PMID- 12888568 TI - Oxygen and iron regulation of iron regulatory protein 2. AB - Iron regulatory protein 2 (IRP2) is a central regulator of cellular iron homeostasis due to its regulation of specific mRNAs encoding proteins of iron uptake and storage. Iron regulates IRP2 by mediating its rapid proteasomal degradation, where hypoxia and the hypoxia mimetics CoCl2 and desferrioxamine (DFO) stabilize it. Previous studies showed that iron-mediated degradation of IRP2 requires the presence of critical cysteines that reside within a 73-amino acid unique region. Here we show that a mutant IRP2 protein lacking this 73-amino acid region degraded at a rate similar to that of wild-type IRP2. In addition, DFO and hypoxia blocked the degradation of both the wild-type and mutant IRP2 proteins. Recently, members of the 2-oxoglutarate (2-OG)-dependent dioxygenase family have been shown to hydroxylate hypoxia-inducible factor-1 alpha (HIF-1 alpha), a modification required for its ubiquitination and proteasomal degradation. Since 2-OG-dependent dioxygenases require iron and oxygen, in addition to 2-OG, for substrate hydroxylation, we hypothesized that this activity may be involved in the regulation of IRP2 stability. To test this we used the 2 OG-dependent dioxygenase inhibitor dimethyloxalylglycine (DMOG) and showed that it blocked iron-mediated IRP2 degradation. In addition, hypoxia, DFO and DMOG blocked IRP2 ubiquitination. These data indicate that the region of IRP2 that is involved in IRP2 iron-mediated degradation lies outside of the 73-amino acid unique region and suggest a model whereby 2-OG-dependent dioxygenase activity may be involved in the oxygen and iron regulation of IRP2 protein stability. PMID- 12888569 TI - Mutant huntingtin promotes the fibrillogenesis of wild-type huntingtin: a potential mechanism for loss of huntingtin function in Huntington's disease. AB - Aggregation of huntingtin (htt) in neuronal inclusions is associated with the development of Huntington's disease (HD). Previously, we have shown that mutant htt fragments with polyglutamine (polyQ) tracts in the pathological range (>37 glutamines) form SDS-resistant aggregates with a fibrillar morphology, whereas wild-type htt fragments with normal polyQ domains do not aggregate. In this study we have investigated the co-aggregation of mutant and wild-type htt fragments. We found that mutant htt promotes the aggregation of wild-type htt, causing the formation of SDS-resistant co-aggregates with a fibrillar morphology. Conversely, mutant htt does not promote the fibrillogenesis of the polyQ-containing protein NOCT3 or the polyQ-binding protein PQBP1, although these proteins are recruited into inclusions containing mutant htt aggregates in mammalian cells. The formation of mixed htt fibrils is a highly selective process that not only depends on polyQ tract length but also on the surrounding amino acid sequence. Our data suggest that mutant and wild-type htt fragments may also co-aggregate in neurons of HD patients and that a loss of wild-type htt function may contribute to HD pathogenesis. PMID- 12888570 TI - Sp1 and Sp3 transcription factors mediate interleukin-1 beta down-regulation of human type II collagen gene expression in articular chondrocytes. AB - Interleukin-1 beta (IL-1 beta) is a pleiotropic cytokine that was shown to inhibit the biosynthesis of articular cartilage components. Here we demonstrate that IL-1 beta inhibits the production of newly synthesized collagens in proliferating rabbit articular chondrocytes and that this effect is accompanied by a decrease in the steady-state levels of type II collagen mRNA. IL-1 beta down regulates COL2A1 gene transcription through a -41/-33 bp sequence that binds a multimeric complex including Sp1 and Sp3 transcription factors. Specificity of IL 1 beta effects on COL2A1 promoter activity was demonstrated in experiments in which transfection of a wild type -50/+1 sequence of COL2A1 promoter as a decoy oligonucleotide abolished the IL-1 beta inhibition of a -63/+47 COL2A1-mediated transcription. By contrast, transfection of the related oligonucleotide harboring a targeted mutation in the -41/-33 sequence did not modify the negative effect the cytokine. Because we demonstrated previously that Sp1 was a strong activator of COL2A1 gene expression via the -63/+1 promoter region, whereas Sp3 overexpression blocked Sp1-induced promoter activity and inhibited COL2A1 gene transcription, we conclude that IL-1 beta down-regulation of that gene, as we found previously for transforming growth factor-beta 1, is mediated by an increase in the Sp3/Sp1 ratio. Moreover, IL-1 beta increased steady-state levels of Sp1 and Sp3 mRNAs, whereas it enhanced Sp3 protein expression and inhibited Sp1 protein biosynthesis. Nevertheless, IL-1 beta decreased the binding activity of both Sp1 and Sp3 to the 63-bp short COL2A1 promoter, suggesting that the cytokine exerts a post-transcriptional regulatory mechanism on Sp1 and Sp3 gene expressions. Altogether, these data indicate that modulation of Sp3/Sp1 ratio in cartilage could be a potential target to prevent or limit the tissue degradation. PMID- 12888571 TI - Critical amino acids in the active site of meprin metalloproteinases for substrate and peptide bond specificity. AB - The protease domains of the evolutionarily related alpha and beta subunits of meprin metalloproteases are approximately 55% identical at the amino acid level; however, their substrate and peptide bond specificities differ markedly. The meprin beta subunit favors acidic residues proximal to the scissile bond, while the alpha subunit prefers small or aromatic amino acids flanking the scissile bond. Thus gastrin, a peptide that contains a string of five Glu residues, is an excellent substrate for meprin beta, while it is not hydrolyzed by meprin alpha. Work herein aimed to identify critical amino acids in the meprin active sites that determine the substrate specificity differences. Sequence alignments and homology models, based on the crystal structure of the crayfish astacin, showed electrostatic differences within the meprin active sites. Site-directed mutagenesis of active site residues demonstrated that replacement of a hydrophobic residue by a basic amino acid enabled the meprin alpha protease to cleave gastrin. The meprin alphaY199K mutant was most effective; the corresponding mutation of meprin betaK185Y resulted in decreased activity toward gastrin. Peptide cleavage site determinations and kinetic analyses using a variety of peptides extended evidence that meprin alphaTyr-199/betaLys-185 are substrate specificity determinants in meprin active sites. These studies shed light on the molecular basis for the substrate specificity differences of astacin metalloproteinases. PMID- 12888572 TI - Functional interaction between the two halves of the photoreceptor-specific ATP binding cassette protein ABCR (ABCA4). Evidence for a non-exchangeable ADP in the first nucleotide binding domain. AB - ABCR, also known as ABCA4, is a member of the superfamily of ATP binding cassette transporters that is believed to transport retinal or retinylidene phosphatidylethanolamine across photoreceptor disk membranes. Mutations in the ABCR gene are responsible for Stargardt macular dystrophy and related retinal dystrophies that cause severe loss in vision. ABCR consists of two tandemly arranged halves each containing a membrane spanning segment followed by a large extracellular/lumen domain, a multi-spanning membrane domain, and a nucleotide binding domain (NBD). To define the role of each NBD, we examined the nucleotide binding and ATPase activities of the N and C halves of ABCR individually and co expressed in COS-1 cells and derived from trypsin-cleaved ABCR in disk membranes. When disk membranes or membranes from co-transfected cells were photoaffinity labeled with 8-azido-ATP and 8-azido-ADP, only the NBD2 in the C-half bound and trapped the nucleotide. Co-expressed half-molecules displayed basal and retinal stimulated ATPase activity similar to full-length ABCR. The individually expressed N-half displayed weak 8-azido-ATP labeling and low basal ATPase activity that was not stimulated by retinal, whereas the C-half did not bind ATP and exhibited little if any ATPase activity. Purified ABCR contained one tightly bound ADP, presumably in NBD1. Our results indicate that only NBD2 of ABCR binds and hydrolyzes ATP in the presence or absence of retinal. NBD1, containing a bound ADP, associates with NBD2 to play a crucial, non-catalytic role in ABCR function. PMID- 12888573 TI - Resistance of the human beta 1-adrenergic receptor to agonist-mediated down regulation. Role of the C terminus in determining beta-subtype degradation. AB - Prolonged agonist stimulation results in down-regulation of most G protein coupled receptors. When we exposed baby hamster kidney cells stably expressing the human beta1-adrenergic receptor (beta 1AR) to agonist over a 24-h period, we instead observed an increase of approximately 30% in both beta 1AR binding activity and immune-detected receptors. In contrast, beta 2AR expressed in these cells exhibited a decrease of > or =50%. We determined that the basal turnover rates of the two subtypes were similar (t(1/2) approximately 7 h) and that agonist stimulation increased beta 2AR but not beta 1AR turnover. Blocking receptor trafficking to lysosomes with bafilomycin A1 had no effect on basal turnover of either subtype but blocked agonist-stimulated beta 2AR turnover. As beta 1AR mRNA levels increased in agonist-stimulated cells, beta 1AR up regulation appeared to result from increased synthesis with no change in degradation. To explore the basis for the subtype differences, we expressed chimeras in which the C termini had been exchanged. Each chimera responded to persistent agonist stimulation based on the source of its C-tail; beta 1AR with a beta 2AR C-tail underwent down-regulation, and beta 2AR with a beta 1AR C-tail underwent up-regulation. The C-tails had a corresponding effect on agonist stimulated receptor phosphorylation and internalization with the order being beta 2AR > beta 1AR with beta 2AR C-tail > beta 2AR with a beta 1AR C-tail > beta 1AR. As internalization may be a prerequisite for down-regulation, we addressed this possibility by co-expressing each subtype with arrestin-2. Although beta 1AR internalization was increased to that of beta 2AR, down-regulation still did not occur. Instead, beta 1AR accumulated inside the cells. We conclude that in unstimulated cells, both subtypes appear to be turned over by the same mechanism. Upon agonist stimulation, both subtypes are internalized, and beta 2AR but not beta 1AR undergoes lysosomal degradation, the fate of each subtype being regulated by determinants in its C-tail. PMID- 12888574 TI - Acid-induced conformational changes in phosphoglucose isomerase result in its increased cell surface association and deposition on fibronectin fibrils. AB - Phosphoglucose isomerase (PGI) is a glycolytic enzyme that exhibits extracellular cytokine activity as autocrine motility factor, neuroleukin, and maturation factor and that has been recently implicated as an autoantigen in rheumatoid arthritis. In contrast to its receptor-mediated endocytosis at neutral pH, addition of 25 microg/ml of either Alexa 568- or FITC-conjugated PGI to NIH-3T3 cells at progressively acid pH results in its quantitatively increased association with cell surface fibrillar structures that is particularly evident at pH 5. A similar pH-dependent cell surface association of PGI is observed for first passage human chondrocytes obtained from osteoarthritic joints. At acid pH, PGI colocalizes with fibronectin fibrils, and this association occurs directly upon addition of PGI to the cells. In contrast to the receptor-mediated endocytosis of PGI, fibril association of 25 microg/ml PGI at pH 5 is not competed with an excess (2 mg/ml) of unlabeled PGI. PGI binding at acid pH is therefore neither saturable nor mediated by its receptor. PGI is enzymatically active as a dimer and we show here by non-denaturing gel electrophoresis as well as by glutaraldehyde cross-linking that it exists at neutral pH in a tetrameric form. Increasingly acid pH results in the appearance of PGI monomers that correlates directly with its enhanced cell surface association. However, glutaraldehyde cross-linked PGI is endocytosed at neutral pH and still exhibits enhanced cell surface binding at pH 5. Circular dichroism analysis revealed pH dependent changes in the near but not the far UV spectra indicating that the tertiary structure of the protein is specifically altered at pH 5. Conformational changes of PGI and exposure of the monomer-monomer interface under acidic conditions, such as those encountered in the synovial fluid of arthritic joints, could therefore result in its deposition on the surface of joints and the induction of an autoimmune response. PMID- 12888575 TI - Constitutive connective tissue growth factor expression in scleroderma fibroblasts is dependent on Sp1. AB - Fibrotic diseases such as scleroderma (systemic sclerosis, SSc) are characterized by an excessive production of extracellular matrix and profibrotic proteins such as connective tissue growth factor (CTGF). In normal dermal fibroblasts, CTGF is not expressed unless induced by proteins such as tumor growth factor-beta (TGFbeta). Conversely, in fibroblasts cultured from fibrotic lesions CTGF mRNA and protein are constitutively expressed, even in the absence of exogenously added TGFbeta. Thus, studying the mechanism underlying CTGF overexpression in SSc fibroblasts is likely to yield valuable insights into the basis of the fibrotic phenotype of SSc and possibly other scarring disease. CTGF overexpression is mediated primarily by sequences in the CTGF promoter. In this report, we identify the minimal promoter element involved with the overexpression of CTGF in SSc fibroblasts. This element is distinct from the element necessary and sufficient for the induction of CTGF expression by TGFbeta in normal fibroblasts. Within this region is a functional Sp1 binding site. Blocking Sp1 activity reduces the elevated, constitutive levels of CTGF promoter activity and protein expression observed in SSc fibroblasts. Relative to those prepared from normal dermal fibroblasts, nuclear extracts prepared from SSc fibroblasts possess increased Sp1 binding activity. Removal of phosphate groups from nuclear extracts enhanced Sp1 binding activity, suggesting that phosphorylation of Sp1 normally reduces Sp1 binding to DNA. Thus, the constitutive overexpression of CTGF in SSc fibroblasts seems to be independent of TGFbeta signaling but dependent at least in part on Sp1. PMID- 12888576 TI - Distinct interaction of versican/PG-M with hyaluronan and link protein. AB - The proteoglycan aggregate is the major structural component of the cartilage matrix, comprising hyaluronan (HA), link protein (LP), and a large chondroitin sulfate (CS) proteoglycan, aggrecan. Here, we found that another member of aggrecan family, versican, biochemically binds to both HA and LP. Functional analyses of recombinant looped domains (subdomains) A, B, and B' of the N terminal G1 domain revealed that the B-B' segment of versican is adequate for binding to HA and LP, whereas A and B-B' of aggrecan bound to LP and HA, respectively. BIAcore trade mark analyses showed that the A subdomain of versican G1 enhances HA binding but has a negligible effect on LP binding. Overlay sensorgrams demonstrated that versican G1 or its B-B' segment forms a complex with both HA and LP. We generated a molecular model of the B-B' segment, in which a deletion and an insertion of B' and B are critical for stable structure and HA binding. These results provide important insights into the mechanisms of formation of the proteoglycan aggregate and HA binding of molecules containing the link module. PMID- 12888577 TI - Cell death caused by selenium deficiency and protective effect of antioxidants. AB - Selenium is an essential trace element and it is well known that selenium is necessary for cell culture. However, the mechanism underlying the role of selenium in cellular proliferation and survival is still unknown. The present study using Jurkat cells showed that selenium deficiency in a serum-free medium decreased the selenium-dependent enzyme activity (glutathione peroxidases and thioredoxin reductase) within cells and cell viability. To understand the mechanism of this effect of selenium, we examined the effect of other antioxidants, which act by different mechanisms. Vitamin E, a lipid-soluble radical-scavenging antioxidant, completely blocked selenium deficiency-induced cell death, although alpha-tocopherol (biologically the most active form of vitamin E) could not preserve selenium-dependent enzyme activity. Other antioxidants, such as different isoforms and derivatives of vitamin E, BO-653 and deferoxamine mesylate, also exerted an inhibitory effect. However, the water soluble antioxidants, such as ascorbic acid, N-acetyl cysteine, and glutathione, displayed no such effect. Dichlorodihydrofluorescein (DCF) assay revealed that cellular reactive oxygen species (ROS) increased before cell death, and sodium selenite and alpha-tocopherol inhibited ROS increase in a dose-dependent manner. The generation of lipid hydroperoxides was observed by fluorescence probe diphenyl-1-pyrenylphosphine (DPPP) and HPLC chemiluminescence only in selenium deficient cells. These results suggest that the ROS, especially lipid hydroperoxides, are involved in the cell death caused by selenium deficiency and that selenium and vitamin E cooperate in the defense against oxidative stress upon cells by detoxifying and inhibiting the formation of lipid hydroperoxides. PMID- 12888578 TI - Structural properties of the chloroplast stromal processing peptidase required for its function in transit peptide removal. AB - The stromal processing peptidase (SPP) catalyzes removal of transit peptides from a diversity of precursor proteins imported into chloroplasts. SPP contains an HXXEH zinc-binding motif characteristic of members of the metallopeptidase family M16. We previously found that the three steps of precursor processing by SPP (i.e. transit peptide binding, removal, and conversion to a degradable subfragment) are mediated by features that reside in the C-terminal 10-15 residues of the transit peptide. In this study, we performed a mutational analysis of SPP to identify structural elements that determine its function. SPP loses the ability to proteolytically remove the transit peptide when residues of the HXXEH motif, found in an N-terminal region, are mutated. Deletion of 240 amino acids from its C terminus also abolishes activity. Interestingly, however, SPP can still carry out the initial binding step, recognizing the C-terminal residues of the transit peptide. Hence, transit peptide binding and removal are two separable steps of the overall processing reaction. Transit peptide conversion to a subfragment also depends on the HXXEH motif. The precursor of SPP, containing an unusually long transit peptide itself, is not proteolytically active. Thus, the SPP precursor is synthesized as a latent form of the metallopeptidase. PMID- 12888579 TI - Cells preconditioned with mild, transient UVA irradiation acquire resistance to oxidative stress and UVA-induced apoptosis: role of 4-hydroxynonenal in UVA mediated signaling for apoptosis. AB - Because 4-hydroxynonenal (4-HNE) has been suggested to be involved in oxidative stress-mediated apoptosis (Cheng, J. Z., Sharma, R., Yang, Y., Singhal, S. S., Sharma, A., Saini, M. K., Singh, S. V., Zimniak, P., Awasthi, S., and Awasthi, Y. C. (2001) J. Biol. Chem. 276, 41213-41223) and UVA irradiation also causes lipid peroxidation, we have examined the role of 4-HNE in UVA-mediated apoptosis. K562 cells irradiated with UVA (3.0 milliwatts/cm2) for 5, 15, and 30 min showed a time dependent increase in 4-HNE levels. As judged by the activation of caspases, apoptosis was observed only in cells irradiated for 30 min. Within 2 h of recovery in normal medium, 4-HNE levels in 5 and 15 min UVA, irradiated cells returned to the basal or even lower levels but in cells irradiated for 30 min, 4 HNE levels remained consistently higher. The cells irradiated with UVA for 5 min and allowed to recover for 2 h in normal medium (UVA-preconditioned cells) showed a remarkable induction of hGST5.8, which catalyzes conjugation of 4-HNE to glutathione (GSH), and RLIP76 (Ral BP-1), which mediates the transport of the conjugate, GS-HNE. In cells irradiated with UVA for 30 min the induction of RLIP76 or hGST5.8 was not observed. The preconditioned cells transported GS-HNE into the medium at a rate about 2-fold higher than the controls and the transport was inhibited (65%) by coating the cells with anti-RLIP76 IgG. Upon treatment with xanthine/xanthine oxidase (XA/XO), 4-HNE, or prolonged UVA exposure, the control cells showed a sustained activation of c-Jun N-terminal kinase (JNK) and apoptosis. However, in the UVA-preconditioned cells, apoptosis was not observed, and JNK activation was inhibited. This resistance of preconditioned cells to XA/XO-, 4-HNE-, or UVA-induced apoptosis could be abrogated when these cells were coated with anti-RLIP76 IgG to block the efflux of GS-HNE. These studies strongly suggest a role of 4-HNE in UVA-mediated apoptosis. PMID- 12888580 TI - Absence of vancomycin-resistant enterococci (VRE) in companion dogs in the conurbation of Parkstad Limburg, The Netherlands. PMID- 12888581 TI - Teicoplanin resistance in methicillin-resistant Staphylococcus aureus in an intensive care unit. PMID- 12888582 TI - Mastic gum has no effect on Helicobacter pylori load in vivo. AB - OBJECTIVE: To determine whether mastic gum suppresses or eradicates Helicobacter pylori infection in humans. PATIENTS AND METHODS: Nine patients with H. pylori infection, and without gastroduodenal ulceration, were recruited from day-case endoscopy lists and treated with mastic 1 g four times daily for 14 days. [13C]Urea breath tests (UBTs) were carried out immediately before, on day 15 and 5 weeks after treatment with mastic. RESULTS: Mastic had no effect on H. pylori status in any of the eight completed patients; all remained H. pylori positive by UBT with no change in delta scores [pre-treatment mean +/- s.e.m. 19.1 +/- 3.7, day 15 (post-treatment) 18.7 +/- 3.8, P = 0.8, paired t-test]. CONCLUSION: Despite reported anti-H. pylori action in vitro, this preliminary study shows that mastic has no effect on H. pylori in humans. PMID- 12888583 TI - Bactericidal activity of levofloxacin against Mycoplasma pneumoniae. PMID- 12888584 TI - Phenotypic and genotypic characterization of antimicrobial resistance in German Escherichia coli isolates from cattle, swine and poultry. AB - OBJECTIVE: Phenotypic and genotypic characterization of the antimicrobial resistance of German Escherichia coli strains isolated during 1999-2001 from cattle, swine and poultry. MATERIALS AND METHODS: Three hundred and seventeen isolates were tested for their resistance to 17 antimicrobial agents by broth microdilution. Resistant strains were screened by molecular methods for resistance genes, integrons and mutations in quinolone-resistance determining regions. RESULTS: Resistance was found in 40% and multiresistance in 32% of the strains. The resistance was significantly higher in isolates from poultry (61%) and swine (60%) than from cattle (25%) (P < 0.01). The most prevalent resistances were to sulfamethoxazole, tetracycline, streptomycin, ampicillin and spectinomycin (30-15%). For each antibiotic, the predominant resistance genes were: ampicillin, blaTEM1-like (92%); chloramphenicol, catA (68%) and cmlA1-like (36%); gentamicin, aac(3)-IV (60%); kanamycin, aphA1 (100%); streptomycin, aadA1 like (61%) and strA/B (59%); sulfamethoxazole, sul2 (66%), sul1 (42%) and sul3 (14%); tetracycline, tet(A) (66%) and tet(B) (42%); and trimethoprim, dfrA1-like (77%), dfrA17 (13%) and dfrA12 (7%). Class 1 integrons were found in 30% of the strains. They carried dfrA1-aadA1a (40%), aadA1a (29%), sat1-aadA1a (16%), dfrA17 aadA5 (11%), oxa1-aadA1a (5%) and dfrA12-aadA2 (3%). Eleven percent of the strains were resistant to nalidixic acid. Of these, 61% presented a reduced susceptibility to ciprofloxacin (MIC = 0.12-2 mg/L) and single mutations in gyrA or gyrA and parC genes, and 39%, full resistance to ciprofloxacin (MIC > or = 4 mg/L) and double and single mutations in gyrA and parC, respectively. CONCLUSION: The study gives baseline information on the magnitude of the resistance problem and its genetic background in contemporary German E. coli from food-producing animals. PMID- 12888585 TI - Comparative activity of telithromycin against typical community-acquired respiratory pathogens. AB - OBJECTIVES: Respiratory tract infections (RTIs) remain a significant cause of morbidity and mortality. Major bacterial pathogens in RTIs, such as Streptococcus pneumoniae, have exhibited increasing resistance to a variety of antibiotics during the past decades. Telithromycin, the first ketolide, was designed especially to overcome this resistance. The present study was conducted to assess the comparative activity of telithromycin against typical RTI pathogens in Austria. METHODS: A total of 1,015 bacterial isolates was tested, including S. pneumoniae, Streptococcus pyogenes, Staphylococcus aureus and Haemophilus influenzae. MICs of the following antimicrobials: penicillin G, ampicillin (for H. influenzae), azithromycin, clarithromycin, erythromycin A and telithromycin were determined using the NCCLS broth microdilution method. RESULTS: Telithromycin showed excellent activity against S. pneumoniae, with 99.8% of all isolates being susceptible. Penicillin remained active with an MIC50 and MIC90 of 0.007 mg/L. Nevertheless, a notable increase in penicillin intermediate-resistant and resistant isolates, from 4.9% in 1996 to the present rate of 10%, was observed. There was also a distinct rise in the resistance levels of S. pneumoniae against the macrolides. All tested isolates of S. pyogenes were susceptible to penicillin and telithromycin, and only low levels of resistance against telithromycin were found in S. aureus (2.2%, MIC90 of 0.5 mg/L). No telithromycin-resistant isolate of H. influenzae could be detected. CONCLUSIONS: This study demonstrates the rising prevalence of resistance among S. pneumoniae not only to penicillin but also to other antimicrobials. It also shows the value of telithromycin as an attractive option for the empirical treatment of community acquired RTIs in an era of widespread antibacterial resistance. PMID- 12888586 TI - Randomized, double-blind study of the clinical efficacy of 3 days of azithromycin compared with co-amoxiclav for the treatment of acute otitis media. AB - BACKGROUND: Compared with 5 days of dosing, a 3 day dosing regimen of azithromycin for treatment of acute otitis media (AOM) may improve compliance, will simplify therapy for the caregiver and, by giving the same total dose as the 5 day regimen, provide more drug when the bacterial burden is highest. METHODS: Children of 6 months-12 years were enrolled if they had had symptoms and signs of AOM for <4 weeks and tympanic membrane effusion by acoustic reflectometry. Eligible children were randomized to azithromycin 10 mg/kg/day x 3 days or co amoxiclav 45 mg/kg/day x 10 days. The primary endpoint was clinical response at day 28. RESULTS: One hundred and eighty-eight children (mean age 3.5 years) were randomized to azithromycin and 185 to co-amoxiclav. At day 10, the clinical success rate was 153/185 (83%) in children treated with azithromycin and 159/181 (88%) in children treated with co-amoxiclav. At day 28, 134/182 (74%) of the children were cured on azithromycin compared with 124/180 (69%) on co-amoxiclav. Also at day 28, signs of AOM, such as abnormal reflectometry (45% versus 59%; P = 0.017), bulging of the eardrum (10% versus 16%; P = 0.059) and loss of tympanic membrane landmarks (11% versus 22%; P = 0.010) were seen less frequently in azithromycin- than co-amoxiclav-treated children, respectively. Adverse events related to therapy were seen in 11% of azithromycin patients compared with 20% on co-amoxiclav (P = 0.014). CONCLUSIONS: Azithromycin given over 3 days is as effective as co-amoxiclav for treatment of AOM, may result in more complete resolution of tympanic membrane disease, and is better tolerated. PMID- 12888587 TI - Effect of antiretroviral protease inhibitors alone, and in combination with paromomycin, on the excystation, invasion and in vitro development of Cryptosporidium parvum. AB - With the spread of the human immunodeficiency virus in the early 1980s, cryptosporidiosis was regarded as an AIDS-defining disease. As an opportunistic pathogen, the intestinal parasite Cryptosporidium parvum became an important cause of chronic diarrhoea, leading to high morbidity and mortality in immunocompromised patients. To date, no effective chemotherapy is available. With the introduction of protease inhibitors (PIs) in highly active antiretroviral therapy (HAART), the incidence of cryptosporidiosis in AIDS patients has declined substantially in western countries. We have therefore tested the effect of five PIs used in HAART on the excystation, invasion and development of the parasite in a cell culture system. The human ileocaecal adenocarcinoma cell line HCT-8 served as a host cell. None of the substances had an effect on the excystation rate, and only nelfinavir moderately, but statistically significantly, inhibited the host cell invasion over a period of 2 h. There were more pronounced inhibitory effects when PIs were present over the total time of intracellular development (48 h). Indinavir, nelfinavir and ritonavir inhibited parasite development significantly. The inhibitory effect was increased when the aminoglycoside paromomycin was combined with the PIs indinavir, ritonavir, and to a lesser extent saquinavir, compared to the PIs alone. PMID- 12888588 TI - Amphotericin B lipid complex versus meglumine antimoniate in the treatment of visceral leishmaniasis in patients infected with HIV: a randomized pilot study. AB - Optimal treatment for HIV-related visceral leishmaniasis (VL) has still to be established. A pilot clinical trial was carried out in 57 HIV-VL coinfected patients to compare the efficacy and safety of amphotericin B lipid complex (ABLC) versus meglumine antimoniate. The patients were randomized to receive either ABLC 3 mg/kg/day for 5 days (ABLC-5, 18 patients), ABLC 3 mg/kg/day for 10 days (ABLC-10, 20 patients) or meglumine antimoniate 20 mg Sbv /kg/day for 28 days (19 patients). Treatment was considered successful if parasites were not detected in a bone marrow aspirate after treatment. Parasitological cure was attained in 33% (95% CI: 13%-59%) of the ABLC-5 group, in 42% (95% CI: 16%-62%) of the ABLC-10 group and in 37% (95% CI: 16%-62%) of the meglumine antimoniate group (P = 0.94). Eight out of 19 patients administered antimoniate discontinued treatment prematurely following serious adverse events, compared with one in the ABLC groups (P = 0.0006). The efficacy of ABLC is similar to meglumine antimoniate, but the severity of toxicity in the treatment of HIV-VL is lower with ABLC. PMID- 12888589 TI - Steady-state plasma and bronchopulmonary characteristics of clarithromycin extended-release tablets in normal healthy adult subjects. AB - OBJECTIVES: The steady-state concentrations of clarithromycin in plasma were compared with concomitant concentrations in epithelial lining fluid (ELF) and alveolar macrophages (AM) obtained from intrapulmonary samples during bronchoscopy and bronchoalveolar lavage (BAL). Concentrations of the major metabolite, 14-hydroxyclarithromycin, were also determined in plasma and AM. MATERIALS AND METHODS: Forty-two healthy, non-smoking adult subjects (age: 18-54 years; 19 females, 23 males) received oral clarithromycin extended-release formulation (1000 mg once daily for five consecutive days). Bronchoscopy and BAL were carried out once in each subject at either 3, 6, 9, 12, 24 or 48 h after the last administered dose of clarithromycin. In addition, three subjects who did not take clarithromycin served as controls and underwent bronchoscopy at 0 h. Drug concentrations in plasma, ELF, and AM were determined by high-performance liquid chromatography. RESULTS: Clarithromycin was extensively concentrated in ELF [range of mean (+/-s.d.) concentrations: 6.38 +/- 3.92 to 11.50 +/- 6.65 mg/L] and AM (127.0 +/- 61.5 to 573.8 +/- 309.3 mg/L) than simultaneous plasma concentration (0.75 +/- 0.31 to 2.22 +/- 0.72 mg/L). The ranges of mean (+/-s.d.) concentrations of 14-hydroxyclarithromycin in plasma and AM were 0.52 +/- 0.29 to 0.80 +/- 0.31 mg/L and 22.1 +/- 13.5 to 49.5 +/- 16.2 mg/L, respectively. CONCLUSIONS: Once-daily dosing of extended-release formulation clarithromycin 1000 mg produced significantly (P < 0.05) higher steady-state concentrations of clarithromycin in ELF (2-14 times) and AM (50-700 times) compared to simultaneous plasma concentrations throughout the 24 h period after drug administration. The 14-hydroxy metabolite of clarithromycin achieved significantly (P < 0.05) higher steady-state concentrations in AM (18-180 times) compared with concurrent plasma concentrations. PMID- 12888590 TI - Human cytomegalovirus resistance to antiviral drugs: diagnosis, monitoring and clinical impact. AB - The incidence of human cytomegalovirus (HCMV) disease in AIDS patients decreased dramatically after the introduction, a few years ago, of highly active antiretroviral combination therapy. As a consequence, the emergence of drug resistant HCMV strains is no longer a major problem in HIV-infected individuals. However, HCMV resistance to antiviral drugs is presently recognized as an emerging problem in transplantation settings. The mechanisms of HCMV drug resistance will be analysed along with the clinical features relevant to the emergence of drug-resistant HCMV strains during antiviral treatment of patients receiving either solid organ or haematopoietic stem cell transplantation. PMID- 12888591 TI - Detection of CTX-M-15 extended-spectrum beta-lactamase in the United Kingdom. PMID- 12888592 TI - Antibiotic activity against urinary tract infection (UTI) isolates of vancomycin resistant enterococci (VRE): results from the 2002 North American Vancomycin Resistant Enterococci Susceptibility Study (NAVRESS). AB - BACKGROUND: The purpose of this study was to assess the prevalence of vancomycin resistant enterococci (VRE) in urinary isolates in North America, and the activity of various antibiotics against VRE. MATERIALS AND METHODS: Twenty-eight medical centres in the United States and 10 centres in Canada assessed the prevalence of VRE in urinary isolates in 2002. Each study site was asked to collect up to a maximum of 50 consecutive VRE (Enterococcus faecium, Enterococcus faecalis only) urinary isolates. Susceptibility was determined by NCCLS broth microdilution. The prevalence of vanA and vanB resistance genotypes was determined by multiplex PCR. RESULTS: From the 28 US medical centres, a total of 697 VRE (616 [88.4%] E. faecium and 81 [11.6%] E. faecalis) were received. Approximately 75% of all VRE (E. faecium and E. faecalis) isolates demonstrated a VanA phenotype (resistance to both vancomycin and teicoplanin). PCR detection of vanA and vanB resistance determinants showed that the vanA genotype was present in 584 of 697 (83.8%) VRE isolates, whereas 113 (16.2%) isolates possessed the vanB gene. The most active agents were linezolid, nitrofurantoin and chloramphenicol, with 0.3%, 0.6% and 2.4% resistance, respectively. The majority (77.8%) of vancomycin-resistant E. faecium isolates displayed the VanA phenotype, and 538 of these 616 (87.3%) isolates were PCR-positive for vanA; the vanB genotype was detected in 78 (12.7%) isolates. Resistance was lowest with linezolid, chloramphenicol and nitrofurantoin at 0.3%, 0.3% and 0.5%, respectively. Only three genetically indistinguishable vanA-positive E. faecium were isolated from the 10 Canadian medical centres. CONCLUSION: VRE urinary isolates are common in the United States, are primarily of the vanA genotype and are very susceptible to linezolid, nitrofurantoin and chloramphenicol. In Canada, VRE urinary isolates remain uncommon. PMID- 12888593 TI - Polymeric carriers for amphotericin B: in vitro activity, toxicity and therapeutic efficacy against systemic candidiasis in neutropenic mice. AB - OBJECTIVE: To study the toxicity and activity of two new amphotericin B formulations: poly(epsilon-caprolactone) nanospheres coated with poloxamer 188 (AmB-NP) and mixed micelles with the same surfactant (AmB-MM). MATERIALS AND METHODS: The toxicity of these formulations was evaluated in erythrocytes, J774.2 macrophages and LLCPK1 renal cells, as well as in mice. Activity was determined in clinical isolates and in neutropenic mice. Mice were made neutropenic with 5 fluorouracil, infected with Candida albicans and treated with the antifungal formulations for three consecutive days. AmB association in cells and accumulation in kidneys and liver of animals was quantified by HPLC. RESULTS: Both formulations decreased between 8- and 10-fold the MIC of the polyene against clinical isolates of C. albicans. However, their activity was lower than or equal to that of AmB-deoxycholate when it was assessed against C. albicans-infected macrophages. When given as a single intravenous dose in mice, AmB-MM and AmB-NP had an LD50 of 9.8 and 18.6 mg/kg, respectively, compared with 4 mg/kg for AmB deoxycholate. Comparison of residual infection burdens in the liver and kidneys showed that AmB-deoxycholate (0.5 mg/kg) was more effective and faster in eradicating yeast cells than polymeric formulations. This fact can be related to a lower AmB accumulation inside macrophages and in liver and kidneys (about 1.5 mg drug/g tissue) of mice, compared with those detected for AmB-deoxycholate (4 mg drug/g). Overall, the efficacy of these formulations at 2 mg/kg was equal to that of AmB-deoxycholate at 0.5 mg/kg. CONCLUSIONS: AmB-MM and AmB-NP decreased the in vivo antifungal activity of AmB, and higher concentrations were therefore necessary to obtain a similar therapeutic effect. However, these higher concentrations were achievable owing to the reduced toxicity of these formulations. PMID- 12888594 TI - Significance of macrolide resistance in Streptococcus pneumoniae. PMID- 12888595 TI - Clonal diversity among erythromycin-resistant beta-haemolytic Streptococcus isolates in La Rioja, Spain. AB - OBJECTIVE: To analyse clonal diversity in 72 erythromycin-resistant beta haemolytic streptococci. METHODS: Clonal relationships were studied for streptococci of groups A (GAS, n = 30), B (GBS, n = 34), C (GCS, n = 4) and G (GGS, n = 4) by means of PFGE. Streptococcal isolates were obtained from a 450 bed hospital in Spain during the following periods: GAS (1996-2001), GBS (1999 2001), GCS and GGS (1997-2000). RESULTS: Twenty-two of the 30 GAS isolates showed the M resistance phenotype and harboured the mef(A) gene. Five unrelated PFGE patterns were identified among these 22 GAS isolates. Sixteen of them, obtained during four different years of the study, showed one of the two predominant PFGE patterns. The remaining eight GAS isolates showed the MLSB resistance phenotype, and four unrelated PFGE patterns were detected. All but one erythromycin resistant GBS showed the MLSB resistance phenotype, and an erm gene was identified in all cases [erm(B) or erm(A)]. Twenty-two unrelated PFGE patterns were demonstrated among 25 GBS with the MLSB resistance phenotype; the remaining eight MLSB GBS isolates could not be typed by PFGE. The eight erythromycin resistant GGS and GCS isolates of this study presented seven unrelated PFGE patterns. GGS and GCS strains showed an MLSB resistance phenotype and had the erm(A) gene. CONCLUSIONS: High clonal diversity was detected in this series of erythromycin-resistant GBS, whereas lower diversity was seen in the GAS isolates. PMID- 12888596 TI - A multicentre comparison of a novel surrogate marker for determining the specific potency of anti-tuberculosis drugs. AB - A model for evaluating the potency of a new anti-tuberculosis drug or a drug combination, based on a decline in the number of viable tubercle bacilli in patient's sputum during 5 days mono-therapy has been reported. One popular measure is based on the analysis of the decline in bacterial counts during the first 48 h of therapy and has been called early bactericidal activity (EBA). Such analyses could detect EBA for only a few drugs and were subject to variations in results obtained in different sites. To address these problems we applied a reiterative exponential decay model to evaluate the data on bacterial counts during 5 days of mono-therapy. The validity of this approach was tested using data from three previously published studies. For patients treated with isoniazid 300 mg daily, the values for the time taken to reduce the viable count by 50% (vt50) measured in days were, from a Kenyan study 0.58 days S.E.M. 0.18, from a Tanzanian study 0.41 days S.E.M. 0.04, and from a United States study 0.55 days s.e.m. 0.12. These differences were not statistically significant (P = 0.77 Kruskal-Wallis non-parametric ANOVA). Mean values of vt50 for all of the major anti-tuberculosis agents showed that there was an overlapping spectrum of activity from isoniazid 300 mg (vt50 0.58 days) to para-amino-salicylic acid (vt50 2.9 days) The variation between column means was greater than could be expected by chance (P = 0.0002 Kruskal-Wallis non-parametric ANOVA). From this, we conclude that the reiterative exponential decay model permits comparison between the data obtained in different centres and would allow the activity of a new drug to be compared with that of the currently available agents. PMID- 12888597 TI - Partial characterization of a transposon containing the tet(A) determinant in a clinical isolate of Acinetobacter baumannii. AB - A genomic library from one clinical isolate of Acinetobacter baumannii was obtained to find genes responsible for tetracycline resistance. Escherichia coli DH5alpha-MDR transformants, selected on Mueller-Hinton agar supplemented with tetracycline 18 mg/L, were thoroughly characterized. In one clone, with an insert of 7,070 bp, it was found that the resistance to tetracycline was mediated by the tet(A) gene (1,200 bp) which encodes a tetracycline efflux pump. This gene was recovered together with tetR(A) (651 bp), the tet(A) repressor. Moreover, the partial sequence (2,008 bp) of a transposase gene, tnpA, and 1,316 bp corresponding to an IS, similar to that described in one strain of Salmonella typhi (IS4321), were found. In this A. baumannii clinical isolate, the tet(A) gene is located in a transposon. The structure of this transposon is similar to that of Tn1721, with the tet(A), tetR(A) and the regions between these genes, being closely related to those of Tn1721. The data indicate horizontal transfer of tetracycline resistance genes between A. baumannii and other genera sharing the same ecological niches. PMID- 12888598 TI - Characterization of the onset and consequences of pneumonia due to fluoroquinolone-susceptible or -resistant Pseudomonas aeruginosa. AB - OBJECTIVES: This study was conducted to identify and compare the microbiological and clinical outcomes among hospitalized adults with pneumonia caused by fluoroquinolone-susceptible or -resistant strains of Pseudomonas aeruginosa. Antibiotic regimens used prior to, as well as those used to treat, the infections were characterized. PATIENTS AND METHODS: This non-randomized multicentre study included 100 consecutively identified patients with pneumonia caused by fluoroquinolone-susceptible (n = 50) or fluoroquinolone-resistant (n = 50) strains of P. aeruginosa. Medical records were examined for demographic, clinical and treatment variables including antibiotics received in the 30 days before the index respiratory or blood culture; AUICs were calculated for each patient using reported or derived MICs. Multivariate logistic and linear regressions were used to identify factors associated with successful clinical and microbiological outcomes. RESULTS: The study population was primarily elderly, frequently in a critical care unit, with low serum albumin and with a high probability of failure and mortality. Patients with pneumonia caused by fluoroquinolone-resistant P. aeruginosa were more likely to have received antibiotics within 7 days before the infection (P = 0.027); the antibiotic regimen was more likely to be of a weak potency (mean AUIC of 58 versus 169, P = 0.001) and to include levofloxacin (P < 0.0001) than what was administered to patients who became infected with a fluoroquinolone-susceptible strain. Regardless of susceptibility, a mean of between 2 and 3 weeks of directed antibiotic therapy was administered to each patient. CONCLUSIONS: Pneumonia caused by fluoroquinolone-resistant P. aeruginosa is frequently associated with prior exposure to levofloxacin. Treatment of P. aeruginosa pneumonia is difficult and usually consists of combination regimens with multiple modifications. PMID- 12888599 TI - Characterization of pentamidine excretion in the isolated perfused rat kidney. AB - OBJECTIVE: To study the renal excretion and kidney accumulation of pentamidine, a potentially nephrotoxic compound, in the isolated perfused rat kidney (IPK). MATERIALS AND METHODS: IPK experiments (3-4 per treatment group) were conducted using male Sprague-Dawley rats (250-350 g). Dose proportionality studies were carried out over a pentamidine dosing range of 80-4000 microg, designed to target initial perfusate concentrations from 1 to 50 microg/mL. Separate interaction experiments were conducted between pentamidine (800 microg) and tetraethylammonium (dose 8000 microg) or dideoxyinosine (dose 80 microg). Inulin was used as a glomerular filtration rate (GFR) marker. Control (drug-naive) perfusions were also carried out. Pentamidine was analysed in perfusate, kidney and urine samples by HPLC. Inulin was measured by a colorimetric method. RESULTS: Pentamidine CLR (1.1 +/- 0.6 to 0.05 +/- 0.03 mL/min) and excretion ratio (3.6 +/ 1.5 to 0.56 +/- 0.15) significantly decreased over the range of doses studied. Significant reductions in viability parameters (GFR, Na reabsorption) were noted in kidneys perfused with high dose pentamidine (4000 microg). Tetraethylammonium co-administration reduced pentamidine renal excretion, resulting in significantly greater kidney accumulation of pentamidine and reduced kidney function. Dideoxyinosine administration had minimal effects on pentamidine disposition. CONCLUSIONS: Pentamidine renal transport involves a combination of mechanisms (filtration, secretion and passive reabsorption). Dose proportionality studies demonstrated non-linear excretion of pentamidine. Inhibition of pentamidine renal clearance by tetraethylammonium was consistent with decreased luminal transport. The detrimental effects of pentamidine on kidney function were the result of significant kidney accumulation of drug. The potential exists for drug-drug interactions between pentamidine and organic cations, increasing the risk of drug induced nephrotoxicity. PMID- 12888600 TI - Pharmacodynamics of linezolid in a clinical isolate of Streptococcus pneumoniae genetically modified to express lux genes. AB - A bioluminescent clinical isolate of Streptococcus pneumoniae was used to test the real-time effects of the oxazolidinone antibiotic, linezolid, on metabolism compared with effects on cell replication. Viable counts and bioluminescence measurements showed that linezolid has little bactericidal effect, which was similar at minimum (6 mg/L), intermediate (13 mg/L) and maximum (20 mg/L) serum concentrations. The post-antibiotic effect, however, was shorter when measured by light output than by viable counts. The results demonstrate that bioluminescence provides a rapid and sensitive means of measuring the effect of antimicrobials on bacterial metabolism, and that the latter recovers earlier than commencement of cell replication after linezolid exposure. PMID- 12888601 TI - Determination of in vitro susceptibility to ABT-492 by BSAC standardized methodology. PMID- 12888602 TI - The nose, nitric oxide, and paranasal sinuses: the outpost of pulmonary antiinfectious defenses? PMID- 12888603 TI - Parasites, allergy, and asthma. PMID- 12888604 TI - Cellular alchemy. PMID- 12888605 TI - A rose by any other name is yet a rose: acute respiratory failure in children. PMID- 12888606 TI - Pro: sleep apnea is an anatomic disorder. PMID- 12888607 TI - Con: sleep apnea is not an anatomic disorder. PMID- 12888610 TI - Principles and practices in the management of obesity. PMID- 12888611 TI - Statement on the care of the child with chronic lung disease of infancy and childhood. PMID- 12888612 TI - Peer review and manuscript triage. PMID- 12888613 TI - Insulin effect on lung diffusion: NO pathway. PMID- 12888614 TI - Lung growth and development after preterm birth: further evidence. PMID- 12888616 TI - The isolated polycystin-1 cytoplasmic COOH terminus prolongs ATP-stimulated Cl- conductance through increased Ca2+ entry. AB - The precise steps leading from mutation of the polycystic kidney disease (PKD1) gene to the autosomal dominant polycystic kidney disease (ADPKD) phenotype remain to be established. Fluid accumulation is a requirement for cyst expansion in ADPKD, suggesting that abnormal fluid secretion into the cyst lumen might play a role in disease. In this study, we sought to establish a link between polycystin 1 (the PKD1 gene product) and ATP-stimulated Cl- secretion in renal tubule cells. To do this, we performed a whole cell patch-clamp analysis of the effects of expression of the isolated cytoplasmic COOH-terminus of polycystin-1 in stably transfected mouse cortical collecting duct cells. The truncated polycystin-1 fusion protein prolonged the duration of ATP-stimulated Cl- conductance and intracellular Ca2+ responses. Both effects were dependent on extracellular Ca2+. It was determined that expression of the truncated polycystin-1 fusion protein introduced, or activated, an ATP-induced Ca2+ entry pathway that was undetectable in transfection control cell lines. Our findings are concordant with increasing evidence for a role of polycystin-1 in cell Ca2+ homeostasis and indicate that dysregulated Ca2+ entry might promote Cl- secretion and cyst expansion in ADPKD. PMID- 12888617 TI - Osmotically inactive skin Na+ storage in rats. AB - Compared with age-matched men, women are resistant to the hypertensive effects of dietary NaCl; however, after menopause, the incidence of salt-sensitive hypertension is similar in women and men. We recently suggested that osmotically inactive Na+ storage contributes to the development of salt-sensitive hypertension. The connective tissues, including those immediately below the skin that may serve as a reservoir for osmotically inactive Na+ storage, are affected by menopause. We tested the hypothesis that ovariectomy (OVX) might reduce osmotically inactive Na+ storage capacity in the body, particularly in the skin. Male, female-fertile, and female OVX Sprague-Dawley (SD) rats were fed a high (8%)- or low (0.1%)-NaCl diet. The groups received the diet for 4 or 8 wk. At the end of the experiment, subgroups received 0.9% saline infusion and urinary Na+ and K+ excretion was measured. Wet and dry weight (DW), water content in the body and skin, total body Na+ (rTBNa+) and skin Na+ (rSKNa+) content were measured relative to DW by desiccation and dry ashing. There were no gender differences in osmotically inactive Na+ storage in SD rats. All SD rats accumulated Na+ if fed 8% NaCl, but rTBNa+ was lower in OVX rats than in fertile rats on a low (P < 0.001)- and a high (P < 0.05)-salt diet. OVX decreased rSKNa+ (P < 0.01) in the rats. A high-salt diet led to Na+ accumulation (DeltaSKNa+) in the skin in all SD rats. Osmotically inactive skin Na+ accumulation was approximately 66% of DeltaSKNa+ in female and 82% in male-fertile rats, but there was no osmotically inactive Na+ accumulation in OVX rats fed 8% NaCl. We conclude that skin is an osmotically inactive Na+ reservoir that accumulates Na+ when dietary NaCl is excessive. OVX leads to an acquired reduction of osmotically inactive Na+ storage in SD rats that predisposes the rats to volume excess despite a reduced Na+ content relative to body weight. PMID- 12888618 TI - Increased expression of cyclooxygenase-1 and -2 in the diabetic rat renal medulla. AB - Alterations in renal prostaglandins (PGs) may contribute to some of the renal manifestations in diabetes leading to nephropathy. PG production is dependent on the activity of cyclooxygenases (COX-1 AND -2) and PG synthases. Our present study investigated levels of these enzymes in streptozotocin-diabetic rats at 2, 4, 6, and 8 wk of diabetes. Immunohistochemical analysis revealed an increase in COX signal in the inner and outer medulla of diabetic rats. This was confirmed by Western blotting, showing up to a fourfold increase in both COX isoforms at 4-6 wk of diabetes. Also, Western blot analysis revealed a sixfold increase in PGE2 synthase expression in the outer medullary region of 6-wk diabetic rats but no difference in the inner medulla. In cultured rat inner medullary collecting duct (IMCD), levels of COX were increased two- to threefold in cells exposed for 4 days to 37.5 mM glucose compared with control of 17.5 mM. While no change in PGE2 synthase levels was noted, PGE2 synthesis was increased. Furthermore, levels of EP1 and EP4 mRNA were increased, as well as a twofold increase in EP4 protein levels. Future studies will determine which COX isoform is contributing to the majority of PGE2 produced in the diabetic IMCD and the significance of these findings to disturbances in IMCD function and to the progression of diabetic nephropathy. PMID- 12888619 TI - AIF: a multifunctional cog in the life and death machine. AB - Mitochondria have a dual role in cellular life and death as life-promoting energy providers and as contributors to programmed cell death (apoptosis). The precise sequence of events resulting in the permeabilization of the mitochondrial membrane and the release of mitochondrial resident proteins remains an actively explored topic. Hansen and Nagley describe results from mammalian cells and from the nematode C. elegans that lead to a feedforward model for mitochondrial destabilization. Furthermore, they describe the mitochondrial and apoptotic functions of several proteins released from mitochondria during progression toward cell death. PMID- 12888620 TI - Colorectal hepatic metastases: detection with SPIO-enhanced breath-hold MR imaging--comparison of optimized sequences. AB - PURPOSE: To compare the accuracy of four breath-hold magnetic resonance (MR) imaging sequences to establish the most effective superparamagnetic iron oxide (SPIO)-enhanced sequence for detection of colorectal hepatic metastases. MATERIALS AND METHODS: Thirty-one patients with colorectal hepatic metastases underwent T1-weighted gradient-echo (GRE) and T2-weighted fast spin-echo (FSE) MR imaging before and after SPIO enhancement. Four sequences were optimized for lesion detection: T2-weighted FSE, multiecho data image combination (MEDIC), T2 weighted GRE with an 11-msec echo time (TE), and T2-weighted GRE with a 15-msec TE. Images were reviewed independently by three blinded observers. The accuracy of each sequence was measured by using alternative free-response receiver operating characteristic analysis. All results were correlated with findings at surgery, intraoperative ultrasonography, or histopathologic examination. Differences between the mean results of the three observers were measured by using the Student t test. RESULTS: Postcontrast T2-weighted GRE sequences were the most accurate and were significantly superior to postcontrast T2-weighted FSE and unenhanced sequences alone (P <.05). For all lesions that were malignant or smaller than 1 cm, respectively, mean accuracies of postcontrast sequences were 0.082 and 0.64 for T2-weighted FSE, 0.90 and 0.78 for MEDIC, 0.92 and 0.80 for GRE with an 11-msec TE, 0.93 and 0.82 for GRE with a 15-msec TE, and 0.81 and 0.62 for unenhanced sequences. CONCLUSION: Optimized SPIO-enhanced T2-weighted GRE combined with unenhanced T2-weighted FSE MR sequences were the most sensitive. Breath-hold FSE postcontrast sequences offer no improvement in sensitivity compared with unenhanced sequences alone. PMID- 12888621 TI - Dual-energy contrast-enhanced digital subtraction mammography: feasibility. AB - A technique for demonstrating breast cancers, dual-energy contrast agent-enhanced digital subtraction mammography, was performed in 26 subjects with mammographic or clinical findings that warranted biopsy. The technique consists of high-energy and low-energy digital mammography after administration of iodinated contrast agent. Weighted subtraction of the logarithmic transform of these images is then performed to obtain an image that preferentially shows iodine. Of the 26 subjects, 13 had invasive cancers. Eleven of these tumors enhanced strongly, one enhanced moderately, and one enhanced weakly. The duct in one patient with ductal carcinoma in situ was weakly enhancing. In the other 12 patients, benign tissue enhanced diffusely in two and weakly focally in two. These results indicate that the technique is feasible and worthy of further study. PMID- 12888622 TI - Caspase cleavage of tau: linking amyloid and neurofibrillary tangles in Alzheimer's disease. AB - The principal pathological features of Alzheimer's disease (AD) are extracellular amyloid plaques and intracellular neurofibrillary tangles, the latter composed of the microtubule-binding protein tau assembled into paired helical and straight filaments. Recent studies suggest that these pathological entities may be functionally linked, although the mechanisms by which amyloid deposition promotes pathological tau filament assembly are poorly understood. Here, we report that tau is proteolyzed by multiple caspases at a highly conserved aspartate residue (Asp421) in its C terminus in vitro and in neurons treated with amyloid-beta (Abeta) (1-42) peptide. Tau is rapidly cleaved at Asp421 in Abeta-treated neurons (within 2 h), and its proteolysis appears to precede the nuclear events of apoptosis. We also demonstrate that caspase cleavage of tau generates a truncated protein that lacks its C-terminal 20 amino acids and assembles more rapidly and more extensively into tau filaments in vitro than wild-type tau. Using a monoclonal antibody that specifically recognizes tau truncated at Asp421, we show that tau is proteolytically cleaved at this site in the fibrillar pathologies of AD brain. Taken together, our results suggest a novel mechanism linking amyloid deposition and neurofibrillary tangles in AD: Abeta peptides promote pathological tau filament assembly in neurons by triggering caspase cleavage of tau and generating a proteolytic product with enhanced polymerization kinetics. PMID- 12888623 TI - Engineered biosynthesis of an ansamycin polyketide precursor in Escherichia coli. AB - Ansamycins such as rifamycin, ansamitocin, and geldanamycin are an important class of polyketide natural products. Their biosynthetic pathways are especially complex because they involve the formation of 3-amino-5-hydroxybenzoic acid (AHBA) followed by backbone assembly by a hybrid nonribosomal peptide synthetase/polyketide synthase. We have reconstituted the ability to synthesize 2,6-dimethyl-3,5,7-trihydroxy-7-(3'-amino-5'-hydroxyphenyl)-2,4-heptadienoic acid (P8/1-OG), an intermediate in rifamycin biosynthesis, in an extensively manipulated strain of Escherichia coli. The parent strain, BAP1, contains the sfp phosphopantetheinyl transferase gene from Bacillus subtilis, which posttranslationally modifies polyketide synthase and nonribosomal peptide synthetase modules. AHBA biosynthesis in this host required introduction of seven genes from Amycolatopsis mediterranei, which produces rifamycin, and Actinosynnema pretiosum, which produces ansamitocin. Because the four-module RifA protein (530 kDa) from the rifamycin synthetase could not be efficiently produced in an intact form in E. coli, it was genetically split into two bimodular proteins separated by matched linker pairs to facilitate efficient inter polypeptide transfer of a biosynthetic intermediate. A derivative of BAP1 was engineered that harbors the AHBA biosynthetic operon, the bicistronic RifA construct and the pccB and accA1 genes from Streptomyces coelicolor, which enable methylmalonyl-CoA biosynthesis. Fermentation of this strain of E. coli yielded P8/1-OG, an N-acetyl P8/1-OG analog, and AHBA. In addition to providing a fundamentally new route to shikimate and ansamycin-type compounds, this result enables further genetic manipulation of AHBA-derived polyketide natural products with unprecedented power. PMID- 12888624 TI - Identification of a G protein-coupled receptor for pheromone biosynthesis activating neuropeptide from pheromone glands of the moth Helicoverpa zea. AB - Pheromone biosynthesis-activating neuropeptide (PBAN), a peptide produced by the subesophageal ganglion, is used by a variety of moths to regulate pheromone production. PBAN acts directly on pheromone gland cells by using calcium and cAMP as second messengers. We have identified a gene encoding a G protein-coupled receptor (GPCR) from pheromone glands of the female moth Helicoverpa zea. The gene was identified based on sequence identity to a group of GPCRs from Drosophila that are homologous to neuromedin U receptors in vertebrates. The full length PBAN receptor was subsequently cloned, expressed in Sf9 insect cells, and shown to mobilize calcium in response to PBAN. This response was dose-dependent (EC50 = 25 nM) with a maximum response at 300 nM and a minimal observable response at 10 nM. Four additional peptides produced by the PBAN-encoding gene were also tested for activity, and it was determined that three had similar activity to PBAN and the other was slightly less active. Peptides belonging to the same family as PBAN, namely pyrokinins, as well as the vertebrate neuromedin U peptide also induced a calcium response. We have identified a GPCR for the PBAN/pyrokinin family of peptides with a known function of stimulating pheromone biosynthesis in female moths. It is related to several receptors from insects (Drosophila and Anopheles) and to neuromedin U and ghrelin receptors from vertebrates. PMID- 12888626 TI - Implications of ventricular arrhythmia vulnerability during murine electrophysiology studies. AB - Programmed ventricular stimulation is being performed for the provocation of ventricular arrhythmias in genetically engineered mice. Despite the high level of interest in this area of translational research, little attention has been given to differentiating between selectivity and specificity of induced ventricular tachycardia (VT) in phenotypically normal mice. We aimed to assess factors that may enhance inducibility of VT in wild-type (WT) mice. In vivo intracardiac electrophysiological studies (EPS) were performed in 230 WT mice of 4 strains. An octapolar electrode catheter was inserted into a jugular vein and advanced to the right atrium and ventricle. Baseline ventricular conduction, refractoriness, and arrhythmia inducibility were assessed using programmed electrical stimulation (PES) and burst pacing. We found that nonsustained VT (> or =4 beats) was inducible in 68/230 (30%) mice. Duration of VT was 1.6 +/- 2.4 s, and the longest episode lasted 24 s. VT inducibility differed by strain and age. Ventricular effective refractory period (VERP) was shorter in mice with inducible VT (44 +/- 12 ms) compared with noninducible mice (61 +/- 16 ms, P < 0.001). VERP increased with age (P < 0.001), albeit with strain-related variability. We conclude that nonsustained VT in WT mice is reproducibly inducible and common. Genetic background variability may predispose certain strains to a higher incidence of arrhythmia induction. EPS methods impact prevalence and specificity of inducible VT. Increased VT inducibility was seen with shorter coupling intervals and application of tightly coupled extrastimuli techniques. These factors should be carefully considered when analyzing PES and burst pacing data in murine models to minimize false positives and optimize accuracy. PMID- 12888625 TI - Selective thyroid hormone receptor-beta activation: a strategy for reduction of weight, cholesterol, and lipoprotein (a) with reduced cardiovascular liability. AB - Few treatments for obesity exist and, whereas efficacious therapeutics for hyperlipidemia are available, further improvements are desirable. Thyroid hormone receptors (TRs) regulate both body weight and cholesterol levels. However, thyroid hormones also have deleterious effects, particularly on the heart. The TR beta subtype is involved in cholesterol lowering and possibly elevating metabolic rate, whereas TR alpha appears to be more important for control of heart rate (HR). In the current studies, we examined the effect of TR beta activation on metabolic rate and HR with either TR alpha 1-/- mice or the selective TR beta agonist KB-141 in mice, rats, and monkeys. 3,5,3'-triiodi-l-thyronine (T3) had a greater effect on increasing HR in WT than in TR alpha-/- mice (ED15 values of 34 and 469 nmol/kg/day, respectively). T3 increased metabolic rate [whole body oxygen consumption (MVO2)] in both WT and TR alpha-/- mice, but the effect in the TR alpha 1-/- mice at the highest dose was half that of the WT mice. Thus, stimulation of MVO2 is likely due to both TR alpha and -beta. T3 had equivalent potency for cholesterol reduction in WT and TR alpha-/- mice. KB-141 increased MVO2 with selectivities of 16.5- and 11.2-fold vs. HR in WT and TR alpha 1-/- mice, respectively. KB-141 also increased MVO2 with a 10-fold selectivity and lowered cholesterol with a 27-fold selectivity vs. HR in rats. In primates, KB 141 caused significant cholesterol, lipoprotein (a), and body-weight reduction (up to 7% after 1 wk) with no effect on HR. TR beta-selective agonists may constitute a previously uncharacterized class of drugs to treat obesity, hypercholesterolemia, and elevated lipoprotein (a). PMID- 12888627 TI - Transcriptional profiling identifies extensive downregulation of extracellular matrix gene expression in sarcopenic rat soleus muscle. AB - The direction of change in skeletal muscle mass differs between young and old individuals, growing in young animals and atrophying in old animals. The purpose of the experiment was to develop a statistically conservative list of genes whose expression differed significantly between young growing and old atrophying (sarcopenic) skeletal muscles, which may be contributing to physical frailty. Gene expression levels of >24,000 transcripts were determined in soleus muscle samples from young (3-4 mo) and old (30-31 mo) rats. Age-related differences were determined using a Student's t-test (alpha of 0.05) with a Bonferroni adjustment, which yielded 682 probe sets that differed significantly between young (n = 25) and old (n = 20) animals. Of 347 total decreases in aged/sarcopenic muscle relative to young muscles, 199 were functionally identified; the major theme being that 24% had a biological role in the extracellular matrix and cell adhesion. Three themes were observed from 213 of the 335 total increases in sarcopenic muscles whose functions were documented in databases: 1) 14% are involved in immune response; 2) 9% play a role in proteolysis, ubiquitin dependent degradation, and proteasome components; and 3) 7% act in stress/antioxidant responses. A total of 270 differentially expressed genes and ESTs had unknown/unclear functions. By decreasing the sample sizes of young and old animals from 25 x 20 to 15 x 15, 10 x 10, and 5 x 5 observations, we observed 682, 331, 73, and 3 statistically different mRNAs, respectively. Use of large sample size and a Bonferroni multiple testing adjustment in combination yielded increased statistical power, while protecting against false positives. Finally, multiple mRNAs that differ between young growing and old, sarcopenic muscles were identified and may highlight new candidate mechanisms that regulate skeletal muscle mass during sarcopenia. PMID- 12888628 TI - Magnesium deficiency induces apoptosis in primary cultures of rat hepatocytes. AB - The effects of extracellular magnesium (Mg) concentration on the rate of apoptosis in rat hepatocytes in primary culture were examined. After overnight attachment, incubations were conducted for up to 72 h in serum-free media containing low (0-0.4 mmol/L), physiological (0.8 mmol/L) or high (2 and 5.6 mmol/L) Mg concentrations. At 72 h, we observed numerous rounded hepatocytes on top of a shrunken cell monolayer at extracellular Mg concentrations < 0.8 mmol/L. These morphological features were associated with Mg-dependent differences in the total protein levels. The various Mg concentrations did not affect DNA synthesis; however, at a concentration < 0.8 mmol/L, the susceptibility of cultured rat hepatocytes to oxidative stress was increased as shown by the reduced glutathione concentration (10.6 +/- 2.8 vs. 37.3 +/- 4.1 nmol/mg protein with 0 and 0.8 mmol/L, respectively; P < 0.05) and increased lipid peroxidation (0.36 +/- 0.03 vs. 0.21 +/- 0.01 nmol malondialdehyde/mg protein with 0 and 0.8 mmol/L, respectively; P < 0.05). Fluorescence microscopy after Hoechst dye staining revealed numerous apoptotic figures in Mg-free monolayers compared with 0.8 and 5.6 mmol/L Mg conditions. These observations were confirmed quantitatively by flow-cytometric analysis after propidium iodide staining. The proportion of subdiploid cells decreased with increasing Mg concentration; for example, it was greater at 72 h in Mg-free cultures (76%) than in cultures containing 0.8 mmol/L or 5.6 mmol/L Mg (28%; P < 0.05). Caspase-3 was highly activated in Mg-free cultures after 48 h of treatment compared with 0.8 and 5.6 mmol/L conditions (P < 0.05). Overall, these results show that extracellular Mg deficiency has a negative effect on the survival of cultured rat hepatocytes by inducing apoptosis; however, supplementation of extracellular Mg did not reduce the spontaneous apoptosis that occurred over time in rat hepatocyte cultures. PMID- 12888629 TI - Octanoate inhibits triglyceride synthesis in 3T3-L1 and human adipocytes. AB - To understand how medium-chain fatty acids (FA) influence lipid metabolism in adipocytes, we studied the effects of octanoate on the oxidation of glucose and endogenous palmitate, cellular O(2) consumption, mitochondrial membrane potential, lipid synthesis from long-chain FA, glucose and lactate. We found that octanoate significantly suppressed the esterification of oleate into triglycerides (TG) in both 3T3-L1 and human adipocytes. Octanoate also significantly suppressed de novo FA synthesis. These effects were associated with octanoate-mediated reductions in the activities of acyl CoA:1,2-diacylglycerol acyltransferase (DGAT) and acetyl CoA carboxylase (ACC). Cells pretreated with octanoate had reduced mRNA levels for a number of lipid metabolism genes, including of DGAT, ACC and stearoyl CoA desaturase-1. On the other hand, octanoate did not acutely perturb cellular O(2) consumption or mitochondrial membrane potential. Together, these results suggest that octanoate affected adipocyte function by reducing TG synthesis but not by enhancing oxidation. PMID- 12888630 TI - Marginal biotin deficiency is teratogenic in ICR mice. AB - The incidence of marginal biotin deficiency in normal human gestation is approximately one in three. In ICR mice, maternal biotin deficiency results in cleft palate, micrognathia, microglossia and limb hypoplasia. However, the relationships among the severity of maternal biotin deficiency, fetal biotin status and malformations have not been reported. This study utilized validated indices of biotin status to investigate the relationships among maternal biotin status, fetal biotin status and the rate of fetal malformations in ICR mice. Biotin status was controlled by feeding diets with varying egg white concentration. In dams and fetuses, biotin status was assessed by hepatic biotin content and hepatic activity of the biotin-dependent enzyme propionyl-CoA carboxylase; in dams, status was also assessed by urinary excretion of biotin and 3-hydroxyisovaleric acid. Malformations were assessed morphologically. Biotin was measured by HPLC/avidin-binding assay. Propionyl-CoA carboxylase (PCC) activity was determined by H(14)CO(3) incorporation. 3-Hydroxyisovaleric acid concentration was determined by GC/MS. Although no overt signs of deficiency appeared, metabolic disturbances caused by biotin deficiency were detectable in dams and fetuses. These disturbances increased with increasing egg white. Fetal biotin status correlated significantly with maternal biotin status (fetal vs. dam hepatic biotin, r = 0.671; fetal vs. dam PCC activity, r = 0.70). The incidences of malformations were strikingly dependent on egg white concentration. We conclude that in ICR mice, marginal maternal biotin deficiency causes fetal biotin deficiency. We speculate that the fetal malformations are primarily the consequence of fetal biotin deficiency. Because murine malformations appeared at degrees of biotin deficiency that are similar to those in human gestation, we speculate that some human fetal malformations may be caused by biotin deficiency. PMID- 12888631 TI - Dietary trans fatty acids alter the compositions of microsomes and mitochondria and the activities of microsome delta6-fatty acid desaturase and glucose-6 phosphatase in livers of pregnant rats. AB - This study was designed to investigate the effects of three diets with different levels of trans fatty acids and the physiologic status on the physicochemical properties and enzymatic activities of liver microsomes and mitochondria. Three groups of 10 female weaning rats each were fed for 10 wk one of three diets differing in their trans fatty acid contents (Control, 0 mol/100 mol total fatty acids; high, 14.5 mol/100 mol; very high, 30 mol/100 mol). At the onset of adult life (10 wk of age), they were mated. Six rats in each group were killed at the end of gestation (Pregnant rat groups). The four remaining pregnant rats continued to receive their experimental diets until weaning of their litters. Six pups from the litters for each group (3 males and 3 females) were selected and fed the same experimental diet as the dams from wk 3 to 10 of age (2nd generation virgin groups) and then killed. Trans fatty acid levels in liver microsomes and mitochondria rose in parallel with the dietary trans fatty acid content, whereas saturated fatty acids dropped in both organelles with increasing trans fatty acids. Pregnant and 2nd generation adult rats fed trans isomers also had lower levels of cholesterol and a lower cholesterol/phosphorus ratio in their liver microsomes compared with controls. A significant interaction between diet and pregnancy was detected in the activities of delta6-desaturase and glucose-6 phosphatase in liver microsomes. Dietary trans fatty acids decreased the activities of both enzymes but only in pregnant rats. No differences in the fluorescence anisotropy of membranes or the enzymatic activities in liver mitochondria were observed. In conclusion, dietary trans fatty acids significantly lowered cholesterol and the cholesterol/phosphorus ratio in liver microsomes. This effect might contribute to low delta6-desaturase and glucose-6 phosphatase activities in liver microsomes of pregnant rats. PMID- 12888632 TI - Olive oil isochromans inhibit human platelet reactivity. AB - The effects of certain polyphenolic compounds in red wine, such as resveratrol and quercetin, have been widely investigated to determine the relationship between dietary phenolic compounds and the decreased risk of cardiovascular diseases. However, the effects of polyphenolic compounds contained in other foods, such as olive oil, have received less attention and little information exists regarding the biological activities of the phenol fraction in olive oil. The aim of this study was to evaluate the antiplatelet activity and antioxidant power of two isochromans [1-(3'-methoxy-4'-hydroxy-phenyl)-6,7-dihydroxy isochroman (encoded L116) and 1-phenyl-6,7-dihydroxy-isochroman (encoded L137)] recently discovered in olive oil and synthesized in our laboratory from hydroxytyrosol. These compounds were effective free radical scavengers and inhibited platelet aggregation and thromboxane release evoked by agonists that induce reactive oxygen species-mediated platelet activation including sodium arachidonate and collagen, but not ADP. Release of tritiated arachidonic acid from platelets was also impaired by L116 and L137. These results indicate that other Mediterranean diet nutraceuticals also exhibit antioxidant activity that could be beneficial in the prevention of vascular diseases. PMID- 12888633 TI - The soybean beta-conglycinin beta 51-63 fragment suppresses appetite by stimulating cholecystokinin release in rats. AB - We previously demonstrated that soybean beta-conglycinin peptone suppresses food intake and gastric emptying by direct action on rat small intestinal mucosal cells to stimulate cholecystokinin (CCK) release. The aim of the present study was to define the active fragment in beta-conglycinin by using synthetic peptides chosen from the sequence of three beta-conglycinin subunits. We selected the fragments that had multiple nonadjacent arginine residues, and investigated their ability to bind to components of the rat intestinal brush border membrane as well as to stimulate CCK release and appetite suppression. The fragment from 51 to 63 of the beta subunit (beta 51-63) had the strongest binding activity. Intraduodenal infusion of beta 51-63 inhibited food intake and markedly increased portal CCK concentration. The threshold concentration of beta 51-63 to affect food intake was 3 micro mol/L. The CCK-A receptor antagonist abolished the beta 51-63-induced suppression of food intake. Three types of smaller fragments of beta 51-63 (beta 51-59, beta 53-63 and beta 53-59) and two types of fragments similar to beta 51-63 in the beta-conglycinin alpha and alpha' subunits (alpha 212-224 and alpha' 230-240) had less binding ability than did beta 51-63. Model peptides constructed with arginine (R) and glycine (G), such as GRGRGRG, had strong binding affinity, but peptides containing a single R or RR did not. These results indicate that the beta-conglycinin beta 51-63 fragment is the bioactive appetite suppressant in beta-conglycinin, and multiple arginine residues in the fragment may be involved in this effect. PMID- 12888634 TI - Zinc deficiency induces oxidative DNA damage and increases p53 expression in human lung fibroblasts. AB - Poor zinc nutrition may be an important risk factor in oxidant release and the development of DNA damage and cancer. Approximately 10% of the United States population ingests <50% of the recommended daily allowance for zinc, a cofactor in proteins involved in antioxidant defenses, electron transport, DNA repair and p53 protein expression. This study examined the effects of zinc deficiency on oxidative stress, DNA damage and the expression of DNA repair enzymes in primary human lung fibroblasts by the use of DNA microarrays and functional assays. Cellular zinc was depleted by 1) growing cells in a zinc-deficient medium and 2) exposuring cells to an intracellular zinc chelator, N,N,N',N'-tetrakis-(2 pyridylmethyl)ethylenediamine. Array data revealed upregulation of genes involved in oxidative stress and DNA damage/repair and downregulation of other DNA repair genes. Zinc deficiency in cells caused an increase in oxidant production (dichlorofluoroscein fluorescence) and a significant induction of single-strand breaks (Comet assay) and p53 protein expression (Western blot analysis). Thus, zinc deficiency not only caused oxidative stress and DNA damage, but also compromised the cells' ability to repair this damage. Zinc adequacy appears to be necessary for maintaining DNA integrity and may be important in the prevention of DNA damage and cancer. PMID- 12888635 TI - Obesity risk is associated with carbohydrate intake in women carrying the Gln27Glu beta2-adrenoceptor polymorphism. AB - Interindividual differences in the response to dietary intake are, in some cases, genotype dependent. Moreover, genotype-environment interactions may appear when the impact of lifestyle factors (e.g., diet) on a phenotype (e.g., BMI > 30 kg/m(2)) differs by genotype. A case-control study (obese subjects vs. normal weight controls) was conducted to assess a possible effect modification on obesity risk of the Gln27Glu polymorphism for the beta(2)-adrenoceptor gene depending on dietary intake. The sample included 159 subjects with BMI > 30 kg/m(2) and 154 controls with BMI < 25 kg/m(2). The allele frequency for the Glu27 polymorphism, as assessed by the polymerase chain reaction-restriction fragment length polymorphism methodology, was 0.40 in cases (obese) and 0.37 in controls (lean), which was similar to that of other Caucasian populations. The dietary intake was estimated by using a previously validated food frequency questionnaire. Obesity incidence was not directly affected by the polymorphism [odds ratio (OR) = 1.40; P = 0.246]. However, a significant interaction (effect modification) between carbohydrate (CHO) intake and the presence of the Glu27 variant in the probability of obesity was apparent. Thus, females with the polymorphism and a higher CHO intake [>49% energy (E)] had a higher obesity risk (OR = 2.56, P = 0.051). The product-term introduced in the logistic model to assess effect modification revealed a marginally significant interaction (P = 0.058) between both factors. Furthermore, a high intake of CHO (E > 49%) was associated with higher insulin levels among women carrying the Gln27Glu polymorphism (P < 0.01). This gene-nutrient interaction emphasizes the importance of examining the outcome of some obesity-related mutations depending on lifestyle (including diet) and may explain the heterogeneity of findings from previous studies. PMID- 12888636 TI - Reduced serum insulin-like growth factor (IGF) I is associated with reduced liver IGF-I mRNA and liver growth hormone receptor mRNA in food-deprived cattle. AB - Nutritional deprivation decreases blood insulin-like growth factor I (IGF-I) concentrations in a variety of species. In this study, we explored the underlying mechanism by determining the effects of food deprivation on the levels of total IGF-I mRNA and total growth hormone receptor (GHR) mRNA, as well as the levels of individual IGF-I mRNA variants and GHR mRNA variants in the liver of steers. Food deprivation for nearly 3 d decreased the levels of serum IGF-I by 63% (P < 0.01), and this decrease was associated with a 75% decrease (P < 0.01) in total IGF-I mRNA in the liver. The food deprivation-induced decrease in liver total IGF-I mRNA was associated with an equivalent decrease in the levels of both class 1 and class 2 IGF-I mRNA. In addition to IGF-I mRNA, food deprivation also decreased the levels of total GHR mRNA in the liver (P < 0.05), and this decrease was associated with a decrease in the liver expression of GHR mRNA variants 1C3 (P < 0.05) and 1A (P = 0.08). Food deprivation did not affect the levels of two other major GHR mRNA variants, 1B and 1C2, in the liver. These results demonstrate that the food deprivation-induced decrease in circulating IGF-I in steers is associated with a coordinate decrease in the expression of different IGF-I mRNA variants and a specific decrease in the expression of GHR mRNA variants 1C3 and 1A in the liver. PMID- 12888637 TI - Low protein intake is associated with reduced hepatic gluconeogenic enzyme expression in rainbow trout (Oncorhynchus mykiss). AB - Our objective was to understand the reasons behind the persistent postprandial hyperglycemia in rainbow trout (Oncorhynchus mykiss). We hypothesized that in this species, high levels of dietary protein could increase the hepatic production of glucose, irrespective of the dietary carbohydrate supply. We fed juvenile rainbow trout four diets containing graded levels of protein for 14 d. Pair-feeding was employed to keep lipid and carbohydrate intakes constant. Six hours after feeding, as postulated, activities and mRNA levels of gluconeogenic enzymes (glucose-6-phosphatase, fructose-1,6-bisphosphatase) increased with increasing dietary protein (P < 0.05). However, in fish with a very low protein intake, there was a very strong increase in plasma glucose (18 mmol/L) that was also associated with a high capacity to store excess glucose as indicated by altered pyruvate kinase activity, glucokinase activity, and hepatic glycogen and fat concentrations (P < 0.05). In conclusion, at the same level of carbohydrate intake, a low dietary protein intake was associated with an unexplained increase in glycemia, which was probably responsible for the decrease in hepatic gluconeogenic enzyme expression. The effect of dietary protein on low carbohydrate utilization in this species remains unclear. PMID- 12888638 TI - Dietary phylloquinone depletion and repletion in older women. AB - Biological markers indicative of poor vitamin K status have been associated with a greater risk for hip fracture in older men and women. However, the dietary phylloquinone intake required to achieve maximal carboxylation of hepatic and extrahepatic vitamin K-dependent proteins is not known. In an 84-d study in a metabolic unit, 21 older (60-80 y) women were fed a phylloquinone-restricted diet (18 micro g/d) for 28 d, followed by stepwise repletion of 86, 200 and 450 micro g/d of phylloquinone. Plasma phylloquinone, urinary gamma-carboxyglutamic acid excretion and gamma-carboxylation of hepatic (prothrombin) and extrahepatic proteins (osteocalcin) decreased in response to phylloquinone restriction (P < 0.001), demonstrating the production of subclinical vitamin K deficiency. The gamma-carboxylation of prothrombin was restored to normal levels in response to phylloquinone supplementation at 200 micro g/d. In contrast, all other biochemical markers of vitamin K status remained below normal levels after short term supplementation of up to 450 micro g/d of phylloquinone. These data support previous observations in rats that hepatic vitamin K-dependent proteins have preferential utilization of phylloquinone in response to phylloquinone dietary restriction. Moreover, our findings suggest that the current recommended Adequate Intake levels of vitamin K (90 micro g/d) in women do not support maximal osteocalcin gamma-carboxylation in older women. PMID- 12888639 TI - Short-term continuous enteral tube feeding schedules did not suppress appetite and food intake in healthy men in a placebo-controlled trial. AB - Tube feeding (TF) provides a model with which to study appetite when nutrient delivery bypasses the upper gastrointestinal (GI) tract and associated cephalic phase response. Nocturnal, diurnal and 24-h TF schedules are used clinically, but their effects on satiety have not been distinguished. This study tested two hypotheses: 1) bypassing the upper GI tract with TF has little satiating effect, and 2) diurnal TF suppresses appetite and food intake more than nocturnal or 24-h schedules. Six healthy men, residing in a metabolic suite, each received three continuous TF schedules (diurnal 12-h, nocturnal 12-h and 24-h; 6.86 +/- 0.51 MJ/d) in random order for 3 d over separate 10-d periods. Two days before and after TF, a placebo feed (<0.4 MJ/d) was given. Weighed measurements of ad libitum food consumption, hourly tracking of appetite and metabolic and hormonal measurements were undertaken. Compared with placebo feeding, there was a nonsignificant reduction in oral intake (1.01-2.49 MJ reduction), little change in appetite sensations with TF and an increase in total energy intake from 14.88 to 20.42 +/- 3.25 MJ. The schedules did not differ although diurnal TF was the most satiating. Changes in a variety of mediators (including leptin, insulin, cholecystokinin, metabolites and respiratory quotient) did not relate to immediate assessments of appetite and food intake. This trial suggests that continuous TF has little effect on satiety in healthy men over a period of 3 d, irrespective of the timing of the schedule. This could not be explained by changes in a variety of metabolic and hormonal signals within the time frame studied. PMID- 12888640 TI - The degree of fat saturation does not alter glycemic, insulinemic or satiety responses to a starchy staple in healthy men. AB - Inclusion of fat reduces the glycemic response to a carbohydate meal, although the effect of different types of fat on glycemic, insulinemic and satiety responses is unclear. Ten healthy men received 50-g carbohydrate portions of mashed potato with isoenergetic amounts of butter (saturated fatty acid), Sunola oil (monounsaturated fatty acid) or sunflower oil (PUFA) and two 50-g glucose loads on separate days. Capillary blood was collected at regular intervals for 2 h. Satiety ratings were assessed by use of a rating scale. The glycemic index (GI), insulin index (II) and satiety index (SI) scores were calculated. Energy intakes from a meal consumed ad libitum at 2 h and for the remainder of the day were quantified. The GI values ranged from 68 +/- 8 to 74 +/- 10 and the II values ranged from 113 +/- 10 to 122 +/- 17, but there was no effect of fat type. SI scores and subsequent energy intake did not differ among the test meals. Substitution of unsaturated fats for saturated fatty acids had no acute benefits on postprandial glycemia, insulin demand or short-term satiety in young men. PMID- 12888641 TI - The Atwater energy equivalents overestimate metabolizable energy intake in older humans: results from a 96-day strictly controlled feeding study. AB - The Atwater energy equivalents of 16.7, 16.7 and 37.7 kJ/g of protein, carbohydrate and fat, respectively, are the standard values used to calculate the macronutrient intakes required to meet a person's metabolizable energy requirement. The aim of this study was to compare in older people the measured metabolizable energy intake (MEI(MEAS)) required to achieve and maintain stable body weights with the MEI estimated using the Atwater energy equivalents (MEI(AT)). During a 96-d (14-wk) strictly controlled dietary period, 11 men and 17 women (55-78 y old) were each provided a MEI(AT) to maintain body weight within +/- 0.5 kg of baseline weight. The MEI(MEAS) was determined retrospectively from the gross energy contents of food, urine and feces samples collected during week 14. Resting energy expenditure was measured using indirect calorimetry. At wk 14, MEI(AT) overestimated MEI(MEAS) by 26%. These results suggest that the Atwater energy equivalent values may overestimate the actual MEI of older people. PMID- 12888642 TI - An overlap of breastfeeding during late pregnancy is associated with subsequent changes in colostrum composition and morbidity rates among Peruvian infants and their mothers. AB - An overlap of breast-feeding and late pregnancy is associated with decreased intake of human milk and reduced infant growth. We evaluated the association of an overlap with macronutrient and immunological components of milk, infant urinary IgA, and infant and maternal morbidity. On d 2 and 1 mo postpartum, staff measured 24-h intake of breast milk and collected samples from 133 Peruvian women; 68 had breast-fed during the last trimester of pregnancy (BFP) and 65 had not breast-fed during pregnancy (NBFP). Data on maternal and infant anthropometry and health were collected for 1 mo. On d 2, lactose and lysozyme concentrations were higher, total lysozyme intake was higher and concentration and total intake of lactoferrin were lower in the BFP than the NBFP group (P < 0.05). The total 1 mo IgA intake was lower among BFP than NBFP infants (P = 0.01). Urinary IgA concentration was correlated with breast milk IgA concentration (r = 0.29; P = 0.01) but not with breast-feeding during pregnancy. An overlap was not associated with diarrhea but BFP infants were 5 times as likely to have a cough for at least 7 d than NBFP infants (P < 0.05). Reported mastitis was rare and occurred only in the NBFP group (P = 0.05). An overlap of breast-feeding and late pregnancy was associated with changes in milk composition, an increased frequency in symptoms of infant respiratory illness but decreased reported mastitis. Further in-depth studies are warranted to determine the cumulative effects associated with a breast-feeding/pregnancy overlap on infant and maternal outcomes. PMID- 12888643 TI - Delayed onset of lactation and risk of ending full breast-feeding early in rural Guatemala. AB - Mothers of infants (n = 328) born between 1996 and 1999 in four Guatemalan communities were interviewed every 2 wk until 6 mo postpartum (pp) to collect prospective breast-feeding data and assess the association between delayed (>3 d pp) onset of lactation (OL) and the risk of ending full breast-feeding. Cox proportional hazards regression was used to examine the association between delayed OL and the hazard of ending full breast-feeding in the first 6 mo, adjusted for potential confounders and effect modifiers. A significant interaction with community was found (community-specific hazard ratios: HR(A) = 2.87, 95% CI = 1.25, 6.60; HR(B) = 3.43, 95% CI = 1.55, 7.59; HR(C) = 0.26, 95% CI = 0.06, 1.14; HR(D) = 1.11, 95% CI = 0.44, 2.77). Supplementation before OL (preonset supplementation) was associated with delayed OL [odds ratio (OR) = 4.87, 95% CI = 2.29, 10.36] and an increased risk of ending full breast-feeding (HR = 1.49, 95% CI = 1.05, 2.11). In the two communities in which mothers who experienced delayed OL had a significantly greater risk of ending full breast feeding than did mothers experiencing normal OL, the association between delayed OL and full breast-feeding was mediated in part by preonset supplementation. Further analyses suggest that this was due to delayed OL leading to nonbreast milk feeds, rather than to introduction of supplements delaying OL. We conclude that some mothers who experience delayed OL are less likely to continue full breast-feeding in the first 6 mo and that further research should examine the contextual factors that modify this relationship. PMID- 12888644 TI - Replacing 40% of dietary animal fat with vegetable oil is associated with lower HDL cholesterol and higher cholesterol ester transfer protein in cynomolgus monkeys fed sufficient linoleic acid. AB - This study was designed to evaluate whether replacing approximately 40 g/100 g dietary animal fat with vegetable oil would improve plasma lipids and lipoproteins when diets contained prudent levels of total saturated acid (SFA), monounsaturated acid (MUFA) and PUFA. Using a cross-over design, male Cynomolgus monkeys (n = 10) were fed purified diets containing a mixture of fats. For the diet based on animal fat (AF-diet), approximately 85 g/100 g of the total fat was derived from pork fat, and approximately 40 g/100 g of this was replaced with olive oil for the vegetable oil-based diet (VO-diet). Thus, the fat content of the VO diet comprised 50% pork fat and 35% olive oil. The remaining 15% of the total fat (for both diets) was safflower oil. Both diets provided approximately 30% of total energy (%en) from fat, <10%en SFA and approximately 6-7%en from PUFA. Monkeys were rotated through two 7-wk feeding periods, during which time plasma lipids and lipoproteins were evaluated. Compared with the AF diet, plasma total cholesterol (TC) concentrations tended to be lower ( approximately 10%) after monkeys consumed the VO diet (3.18 +/- 0.83 vs. 3.52 +/- 0.93 mmol/L, P = 0.099), and this was due entirely to a significant 12% reduction in HDL cholesterol (1.53 +/- 0.41 vs. 1.73 +/- 0.47, mmol/L, P = 0.0009). Although plasma lipoprotein compositional analyses revealed no significant differences in either lipoprotein composition or the estimated particle diameters, the measurement of cholesterol ester transfer protein (CETP) using (3)H-cholesterol ester-labeled HDL revealed that the lower HDL cholesterol (HDL-C) when monkeys consumed the VO diet was associated with a 31% increase in transfer (P = 0.04). However, despite the changes in HDL-C, the TC/HDL-C ratio did not differ between monkeys after the two diet treatments. Regression analyses of data from these monkeys revealed a significant correlation between the dietary 16:0/18:2 ratio and plasma HDL-C. These data suggest that within the context of currently recommended prudent diets, it may be possible to manipulate HDL-C beneficially. Whether a similar effect would occur in humans warrants investigation. PMID- 12888645 TI - Choline uptake in human intestinal Caco-2 cells is carrier-mediated. AB - The objective of the current investigation was to examine the transport characteristics of choline, an endogenous quaternary ammonium compound, into human intestinal Caco-2 cells; the transport of choline has not been characterized in human intestine. The cellular accumulation of choline was independent of an inwardly directed Na(+) gradient and demonstrated temperature dependence and saturability. Using the initial uptake rates, choline accumulation was best characterized by a Michaelis-Menten equation and a diffusion component with a K(m) and V(max) of 110 +/- 3 micro mol/L and 2800 +/- 250 pmol/(mg protein. 10 min), respectively. Choline uptake was significantly inhibited by an excess of choline itself and by hemicholinium-3, a structural analog of choline. However other hydrophilic organic cations, such as tetraethylammonium (TEA) and N methylnicotinamide (NMN), did not affect choline uptake in Caco-2 cells. Additionally, two typical p-glycoprotein substrates, daunomycin and verapamil, both inhibited choline accumulation. However the opposite was not true: choline did not inhibit DNM accumulation in Caco-2 cells. These results indicate the presence of a carrier-mediated transport system for choline in Caco-2 cells. The substrate specificity of this carrier is unlike that seen in the rat intestinal epithelium, and the human transport protein is distinct from those for TEA and NMN. P-glycoprotein substrates may inhibit choline uptake through specific or nonspecific interactions with the choline transporter. PMID- 12888646 TI - Hydroxytyrosol excretion differs between rats and humans and depends on the vehicle of administration. AB - Interest in the in vivo biological activities of olive oil phenolics is rapidly growing, and different models and vehicles of administration are used worldwide. Matters of practicality determine the use of rats rather than humans as the model of choice. Also, growing interest in nutraceuticals is leading to the formulation of compounds containing olive oil phenols. In this study, we compared metabolism and urinary excretion of hydroxytyrosol [(HT), the most representative phenol of olive oil] between rats and humans by evaluating excretion of HT and its major metabolite, homovanillyl alcohol. Also, we compared human excretion of HT when consumed as a natural component of extra virgin olive oil, when added to refined olive oil, or when added to yogurt (as an approximation of functional food). Urinary excretion of HT was greater in humans than in rats, a species with a high basal excretion of HT and its metabolites. The high (234% of HT administered) excretion of free HT suggests that hydrolysis of oleuropein administered in humans (still an unresolved issue) occurs in vivo. Moreover, human HT excretion was much higher after its administration as a natural component of olive oil (44.2% of HT administered) than after its addition to refined olive oil (23% of HT administered) or yogurt (5.8% of dose or approximately 13% of that recorded after virgin olive oil intake). These data suggest that the rat is not the appropriate model for the study of HT metabolism and that HT-containing functional foods should be carefully formulated. PMID- 12888647 TI - Taurine prevents hypercholesterolemia in ovariectomized rats fed corn oil but not in those fed coconut oil. AB - We studied whether the type of dietary fatty acid influences the preventive effect of taurine on the ovarian hormone deficiency-induced increase in plasma cholesterol concentration in 6-mo-old ovariectomized rats. Rats were fed one of the following four diets for 28 d: purified diets based on corn oil, which is rich in linoleic acid, with or with out taurine (50 g/kg) or purified diets based on coconut oil, which is rich in lauric and myristic acids, with or without taurine. Body mass gain, food intake, liver weight and plasma apolipoprotein (apo) A-I, apo B, LDL and VLDL concentrations were not affected by the diets. On the other hand, taurine lowered the plasma total cholesterol concentration (P < 0.02) in rats fed corn oil, but not in those fed coconut oil. In rats fed both types of oils, taurine increased the LDL receptor mRNA level (P < 0.01), hepatic cholesterol 7alpha-hydroxylase activity (P < 0.01) and fecal bile acid excretion (P < 0.01). Taurine increased the HMG-CoA reductase mRNA level (P < 0.02) in the liver of rats fed coconut oil, but not in those fed corn oil. Taurine increased liver total lipid (P < 0.05) and triglyceride (P < 0.05) concentrations in rats fed corn oil, but not in those fed coconut oil. These results indicate that the effect of taurine on ovarian hormone deficiency-induced changes in cholesterol metabolism is influenced by the type of dietary fatty acids. PMID- 12888648 TI - Black tea prevents cigarette smoke-induced oxidative damage of proteins in guinea pigs. AB - Cigarette smoke (CS) causes oxidative damage and tea polyphenols have strong antioxidant properties. Therefore, we studied the effect of a black tea (BT) infusion on CS-induced oxidative damage of proteins both in vitro and in vivo. In the in vitro experiment, bovine serum albumin (BSA) or a guinea pig tissue microsomal suspension was incubated with an aqueous extract of CS (CS-solution) in the presence or absence of the BT infusion. Protein oxidation was measured by immunoblotting of the dinitrophenylhydrazone derivatives of the protein carbonyls followed by densitometric scanning. Protein degradation was assessed by SDS-PAGE. BT prevented (P < 0.05) CS-induced oxidation of BSA and oxidative degradation of guinea pig lung, liver and heart microsomal proteins. This was also observed when the BT infusion was replaced by its components, i.e, flavonols, theaflavins, thearubigins and catechins. BT prevented microsomal protein degradation by inhibiting oxidative modification of the proteins. The antioxidant effect of BT was similar to that of green tea. In the in vivo experiment, partially ascorbate deficient guinea pigs were subjected to CS exposure from 5 cigarettes/(guinea pig. d) for 7 d and given water or the BT infusion (20 g/L) to drink. Guinea pigs exposed to CS and given water had extensive oxidation accompanied by 39, 40 and 30% losses (P < 0.05) of microsomal proteins of lung, liver and heart, respectively. However, when the CS-exposed guinea pigs consumed the BT infusion instead of water, the oxidation of microsomal proteins was reduced (P < 0.05) approximately 90, 97 and 70% in lung, liver and heart, respectively. Protein loss was reduced (P < 0.05) approximately 92, 98 and 90% in lung, liver and heart, respectively. The results, if extrapolated to humans, would indicate that regular intake of tea may protect smokers from CS-induced oxidative damage and consequent degenerative diseases. PMID- 12888649 TI - Vitamin A depletion induced by cigarette smoke is associated with the development of emphysema in rats. AB - We showed previously that vitamin A deficiency per se causes emphysema. Benzo(a)pyrene, a constituent in cigarette smoke, induces vitamin A depletion when administered to rats; therefore, we tested the hypothesis that cigarette smoke induces vitamin A depletion, which is associated with the development of emphysema. Male weanling rats were fed a purified AIN-93G diet and divided into two groups. The experimental group was exposed to cigarette smoke from 20 nonfiltered commercial cigarettes/d for 5 d/wk, whereas the control group was exposed to air. After 6 wk, tissues were collected for histological and biochemical analyses. Retinol levels were measured in serum, lung and liver. The trachea, lung and liver were examined for histological changes. Vitamin A levels decreased significantly in serum, lung and liver of smoke-treated rats. Histological examination revealed the presence of interstitial pneumonitis along with severe emphysema. There was a significant inverse relationship between vitamin A concentration in the lung and the severity of emphysema (r = -0.69 and P < 0.03). Detachment or hyperplasia (and metaplasia) of the tracheal epithelium and liver vacuole formation also were evident in the smoke-treated rats. The results of this research indicate that exposure to cigarette smoke induces vitamin A depletion in rats, which is associated with the development of emphysema. PMID- 12888650 TI - Iron-deficient mice fail to develop autoimmune encephalomyelitis. AB - Determinations of the effects of iron status on the immune system are complicated by the fact that microorganisms and immune cells both utilize iron. To determine the role of iron in immune function, we utilized a model [experimental autoimmune encephalomyelitis (EAE)] in which a strong antigen-specific CD4+ T-cell response develops in the absence of infection. EAE is an autoimmune disease frequently used as a model for the human disease multiple sclerosis (MS). EAE was induced in B10.PL mice fed low iron (1 mg/kg), normal iron (10 mg/kg) or high iron (160 mg/kg) diets that were replete in all other nutrients. Liver iron measurements verified iron status, i.e., low iron mice had 1.9 micro mol/g tissue, normal iron mice, 3.27 micro mol/g tissue and high iron mice, 5.35 micro mol/g tissue. EAE symptoms were most severe in normal iron mice, and EAE did not develop in low iron mice. The incidence of EAE was 71% in normal iron mice, 62% in iron overloaded mice and 0% in iron-deficient mice. Two of seven mice in the normal iron group developed severe EAE and were euthanized. None of the iron-overloaded mice developed severe EAE. Other measures of EAE severity were similar in the normal and iron-overloaded mice. The data suggest that iron deficiency provides protection from the development of EAE and that iron excess with its potential contribution to free radical formation was not an important factor. The mechanism of EAE inhibition in iron-deficient mice likely involves the delivery and metabolism of iron for optimal CD4+ T-cell development. PMID- 12888651 TI - Total subdiaphragmatic vagotomy does not suppress high protein diet-induced food intake depression in rats. AB - This study was undertaken to determine whether the subdiaphragmatic vagus nerve is involved in the depression of food intake induced by the ingestion of a high protein diet (P50) in rats. After total subdiaphragmatic vagotomy (Vago group) or sham surgery (Sham group), rats consumed the control diet for a 2-wk recovery period and then both groups consumed the high protein diet for 16 d. Daily food intake, meal pattern analysis and behavioral satiety sequence were measured. Total subdiaphragmatic vagotomy did not modify the daily intake of the control diet or suppress the dramatic depression in food intake produced by acute transition to a high protein diet. However, the daily intake of a high protein diet was slightly reduced under acute conditions or even after adaptation (P < 0.005). Analysis of meal parameters and the behavioral satiety sequence after adaptation indicated no major metabolic distress. In conclusion, these results suggest that the subdiaphragmatic vagus nerve does not constitute an obligatory pathway for the transfer of information to the brain, resulting in a depression of high protein diet intake. In contrast, a defect in this visceral regulating system could reinforce the metabolic-associated food intake depression signal. PMID- 12888652 TI - Serum total homocysteine concentrations in children and adolescents: results from the third National Health and Nutrition Examination Survey (NHANES III). AB - Although the elevation of circulating total serum homocysteine (tHcy) concentration in a fasting state is associated with an increased risk of occlusive vascular disease in adults, the implications of elevated levels in children are not known. The goals of this study were to describe the distribution of tHcy among a representative sample of children and adolescents in the United States, and to test for differences in tHcy among sex, age and race-ethnicity categories. Using surplus sera from Phase 2 of the third National Health and Nutrition Examination Survey, we measured tHcy for a nationally representative sample of 942 boys and 1085 girls aged 4-19 y. The age-adjusted geometric mean tHcy concentrations were 6.2 and 5.8 micro mol/L in non-Hispanic Caucasian boys and girls, 6.4 and 6.1 micro mol/L in non-Hispanic African-American boys and girls, and 6.4 and 5.5 micro mol/L in Mexican American boys and girls, respectively. A significant interaction between age and sex (P < 0.01) reflected the divergence of tHcy concentrations at about age 10 y, with higher concentrations in boys than in girls throughout adolescence. These first data on homocysteine concentrations in a nationally representative sample of American youth suggest that sexual dimorphism of tHcy concentrations occurs earlier, at approximately 10 y of age, than previously reported on the basis of smaller nonrepresentative samples. Improved understanding of the determinants of levels during growth and development may provide important clues to the etiology of adult disease. PMID- 12888653 TI - Multivitamin supplements are inversely associated with risk of myocardial infarction in men and women--Stockholm Heart Epidemiology Program (SHEEP). AB - Epidemiologic data relating multivitamin supplement use to the risk of cardiovascular disease are sparse and inconsistent. We examined the association between self-selected use of low dose multivitamin supplements and the risk of myocardial infarction (MI). Our results are based on data from a large population based, case-control study of subjects aged 45-70 y residing in Sweden, a country in which consumption of fruits and vegetables is relatively low and foods are not fortified with folic acid. The study included 1296 cases (910 men, 386 women) with a first nonfatal MI and 1685 controls (1143 men, 542 women) frequency matched to the cases by sex, age and hospital catchment area. Odds ratios (OR) and 95% CI were calculated from unconditional logistic regression models. Among controls, 57% of the women and 35% of the men used dietary supplements; corresponding figures for the cases were 42 and 27%, respectively. Of those taking supplements, 80% used multivitamin preparations. After adjustment for major cardiovascular risk factors, the OR of MI comparing regular users of supplements with nonusers were 0.79 (95% CI 0.63-0.98) for men and 0.66 (95% CI 0.48-0.91) for women. This inverse association was not modified by such healthy lifestyle habits as consumption of fruits and vegetables, intake of dietary fiber, smoking habits and level of physical activity, although never smoking appeared to outweigh the association in women. Findings from this study indicate that use of low dose multivitamin supplements may aid in the primary prevention of MI. PMID- 12888654 TI - Alcohol drinking patterns differentially affect central adiposity as measured by abdominal height in women and men. AB - Alcohol drinking in light-to-moderate amounts has been associated with reduced coronary heart disease (CHD) risk. However, there is evidence that the way people consume alcohol (drinking pattern) may affect risk. Central adiposity, a known CHD risk factor may be one mechanism in the pathway between alcohol consumption and CHD risk. Our study examined whether various drinking patterns differentially affect fat distribution, particularly abdominal fat in women and men. In a randomly selected population-based cohort (n = 2343), 35-79 y old, we assessed drinking pattern as reported for the past 30 d, including beverage type and amount, frequency of consumption, percentage of time drinking while eating and number of drinks consumed/drinking day. Central adiposity was determined using an abdominal caliper to measure supine height of the abdomen. Current drinkers tended to have smaller abdominal heights than nondrinkers (women, P < 0.0001; men, P = 0.0559). For drinking pattern, frequency was inversely associated, but drinking intensity (drinks/drinking day) was positively associated with central adiposity in women (P trend for frequency, 0.0007; intensity, 0.0010) and men (P trend for frequency, 0.0005; intensity, 0.0004), even when age, education, physical activity, smoking status and amount of alcohol (g) were included in the models. When frequency and intensity were considered together, daily drinkers of <1 drink/drinking day had the smallest mean abdominal height measures with the largest measures in less than weekly drinkers who consumed 4 or more drinks/drinking day. These results support the hypothesis that drinking pattern affects the distribution of body fat, an important CHD risk factor. PMID- 12888655 TI - The reliability of ten-year dietary recall: implications for cancer research. AB - Remote dietary intakes may be more important than recent diet in the etiology of cancer because of the long latency in cancer development. We examined the reliability of remote dietary recall over 10 y. Subjects were 56 adults participating in a cancer prevention trial in Western Australia. All subjects completed a 28-d diet record (DR) in 1991. A food-frequency questionnaire (FFQ) modified to ask respondents about their diet 10 y earlier was sent to each subject for completion in 2001. Remote intakes recalled from 10 y earlier using the FFQ were compared with the DR using the limits of agreement (LOA) method and Pearson correlation coefficients. Mean intakes of most nutrients did not differ between dietary methods. The LOA indicated that the FFQ could under- or overestimate DR estimates by >/=50%. For many nutrients, agreement between methods depended on the magnitude of intake. Pearson's correlation coefficients ranged from 0.02 for retinol to 0.66 for alcohol. These findings are similar to those of other studies that examined the reliability of recent and remote dietary intakes. They also show that using this FFQ, remote diet recalled from 10 y earlier may be as reliable as recent dietary recall. PMID- 12888656 TI - Low concentrations of quercetin and ellagic acid synergistically influence proliferation, cytotoxicity and apoptosis in MOLT-4 human leukemia cells. AB - Little information is available regarding possible synergistic or antagonistic biochemical interactions among polyphenols contained in fruits and vegetables. Identifying potential interactions among these compounds may help to define the efficiency of polyphenol-containing foods in cancer prevention as related to structure-function activity of the compounds. The objective of this study was to investigate interactions between quercetin and ellagic acid, two polyphenolics that are present predominantly in small fruits, on cell death and proliferation related variables in the MOLT-4 human leukemia cell line. Assays were performed to determine cell cycle kinetics, proliferation, apoptotic DNA-fragmentation and caspase-3-activity after 12, 24 and 48 h. Ellagic acid significantly potentiated the effects of quercetin (at 5 and 10 micro mol/L each) in the reduction of proliferation and viability and the induction of apoptosis. Significant alterations in cell cycle kinetics were also observed. The synergy was confirmed by an isobolographic analysis of the cell proliferation data. The interaction of ellagic acid and quercetin demonstrated an enhanced anticarcinogenic potential of polyphenol combinations, which was not based solely on the additive effect of individual compounds, but rather on synergistic biochemical interactions. PMID- 12888657 TI - Conjugated linoleic acid downregulates insulin-like growth factor-I receptor levels in HT-29 human colon cancer cells. AB - Conjugated linoleic acid (CLA) has chemoprotective properties in a variety of experimental cancer models. We have previously observed that dietary CLA inhibits colon tumorigenesis induced by 1,2-dimethylhydrazine in rats. In addition, our in vitro studies have shown that CLA inhibits DNA synthesis and induces apoptosis in HT-29 cells, the human colon adenocarcinoma cell line. The insulin-like growth factor (IGF) system regulates the growth of HT-29 cells by an autocrine mechanism. The present study examined whether the growth inhibitory effect of CLA is related to changes in the IGF system in HT-29 cells. To determine whether CLA inhibits IGF-II production, HT-29 cells were incubated in serum-free medium in the presence of various concentrations of CLA. CLA decreased protein levels of both mature and pro IGF-II and IGF-II transcripts. Whereas exogenous IGF-I and IGF-II produced an increase in cell number, neither IGF-I nor IGF-II counteracted the negative growth regulatory effect of CLA. Reverse transcriptase-polymerase chain reaction and Western blot analysis of total cell lysates revealed that CLA decreased IGF-I receptor (IGF-IR) transcript and protein levels in a dose dependent manner. Immunoprecipitation/Western blot studies revealed that CLA inhibited IGF-I-induced phosphorylation of IGF-IR and insulin-receptor substrate (IRS)-1, recruitment of the p85 regulatory subunit of phosphatidylinositol 3 kinase (PI3K) to IGF-IR, IGF-IR-associated PI3K activity, and phosphorylated Akt and extracellular signal-regulated kinase (ERK)-1/2 levels. In conclusion, the inhibition of cell proliferation and induction of apoptosis by CLA in HT-29 cells may be mediated in part by its ability to decrease IGF-II synthesis and to downregulate IGF-IR signaling and the PI3K/Akt and ERK-1/2 pathways. PMID- 12888658 TI - Down-regulation of proliferating cell nuclear antigen gene expression occurs during cell cycle arrest induced by human fecal water in colonic HT-29 cells. AB - Cancer is a disease in which the cell cycle is altered, and the elucidation of the mechanisms by which constituents of human fecal water influence the cell cycle can lead to noninvasive measurement of colon cancer risk. The purpose of the present study was to investigate the effect of human fecal water on HT-29 cell-cycle progression with sodium selenite as a reference for comparison. Both human fecal water (2.5-5.0%) and selenite (3-4 micro mol/L) significantly inhibited cell growth. Cell-cycle analysis revealed that human fecal water decreased the proportion of S + G2 phase cells and increased that of G1 phase cells. In contrast, selenite decreased G1 phase cells and increased proportions of S and G2 phase cells. Both 5% human fecal water and 4 micro mol/L selenite greatly increased the mRNA level of the cyclin-dependent kinase inhibitor gene p21(waf1). Interestingly, the mRNA levels of cyclin A and proliferating cell nuclear antigen (PCNA) were dramatically decreased by 69 and 62%, respectively, in HT-29 cells treated with fecal water but not selenite. In contrast, the mRNA level of DNA damage-inducible transcript 1, gadd45, was significantly increased by 2.28-fold in HT-29 cells treated with selenite but not fecal water. Furthermore, a PCNA gene promoter was cloned into a luciferase reporter construct and its activity was significantly reduced in a dose-dependent manner in cells treated with fecal water but not selenite. Collectively, these results suggest that human fecal water and selenite can differentially induce growth arrest genes, and that PCNA gene expression is uniquely and highly sensitive to human fecal water. PMID- 12888660 TI - Electrophoretic patterns of proteins from Paragonimus westermani in early developmental stages. AB - In order to observe the protein compositions of soluble extracts of P. westermani, and their changes during early developmental stages, the crude saline extracts of 4, 6, 8, 10 and 12 week-old worms which were harvested from experimentally infected dogs were analysed by disc-PAGE. The results were as follows: A total of 15 bands were identified from electrophoregrams of respective developmental stages. Of them, 5 bands were recognized throughout the developmental stages. The number and protein amount of identified bands changed according to the worm development from 4 weeks to 12 weeks. However, the banding patterns of 4 and 6 week-old worms and 8 and 10 week-old worms were similar each other. Of 15 identified bands, band 1 was recognized only in 12 week-old worms whereas bands 3, 4, 8, 9, 10, 11 and 15 gradually lowered their amount according to development to disappear in 12 week-old. In addition, band 5 became a major band in 12 week-old while band 6 turned to a minor band at tha same age. The possible relations of changing patterns of protein bands with worm development were discussed. PMID- 12888659 TI - The activities of antioxidant nutrients in human plasma depend on the localization of attacking radical species. AB - The oxidation of endogenous antioxidant nutrients in human plasma was determined to examine their activities against free radicals generated in the aqueous and lipid compartments of plasma. Free radicals were induced at a constant rate in the aqueous compartment by the hydrophilic radical generator, 2,2'-azobis-(2 amidinopropane)dihydrochloride (AAPH; 10-20 mmol/L) and in the lipid compartment by the lipophilic radical generator, 2,2'-azobis(4-methoxy-2,4 dimethylvaleronitrile) (MeO-AMVN; 1-2 mmol/L). The depletion of endogenous plasma antioxidant nutrients (lutein, cryptoxanthin, beta-carotene, lycopene, alpha tocopherol, ascorbic acid, uric acid) was determined after incubation with either AAPH or MeO-AMVN at 37 degrees C using HPLC. The oxidation of the aqueous and lipid compartments of plasma was selectively monitored by a fluorimetric method using either the hydrophilic probe, 2',7'-dichlorodihydrofluorescein (DCFH) or the lipophilic probe, 4,4-difluoro-5-(4-phenyl-1,3-butadienyl)-4-bora-3a, 4a diaza-s-indacene-3-undecanoic acid (BODIPY 581/591). When plasma was incubated with AAPH, the rates of consumption of the antioxidant nutrients were as follows: ascorbic acid > alpha-tocopherol > uric acid > lycopene > lutein > cryptoxanthin > beta-carotene. When plasma was incubated with MeO-AMVN, alpha-tocopherol and carotenoids were depleted at similar rates and ahead of the major water-soluble antioxidants. Our study indicates that the antioxidant nutrients present in both the lipid and aqueous compartments can remove free radicals generated in plasma, and their activity depends on the localization of the attacking radical species. PMID- 12888661 TI - Heterophyid trematodes (Heterophyopsis continua, Pygidiopsis summa and Heterophyes heterophyes nocens) from domestic cats in Korea. AB - During 8 april 1983 to 9 April 1985, a total of 181 domestic cats (Felis catus domestica Linnaeus) of unknown localities in Korea were purchased at Seoul Chung ang market, and the internal organs were dissected for helminth infections. In the results, three kinds of trematodes of family Heterophyidae (Ohdner, 1914) were obtained from small intesine of domestic cats and identified as follows: five adult worms of Heterophyopsis continua (Onji et Nishio, 1916) from three hosts, two hundreds and twenty three adult worms of Pygidiopsis summa Onji et Nishio, 1916 from seven hosts and twenty six adult worms of Heterophyes heterophyes nocens (Onji et Nishio, 1916) from three hosts. Among them, Heterophyopsis continua and Pygidiopsis summa had not been proved as natural infectios of cats in Korea. PMID- 12888662 TI - In situ posture of anterior body of Metagonimus yokogawai in experimentally infected dog. AB - The in situ posture of anterior body of Metagonimus yokogawai was observed in experimental metagonimiasis of dog. The metacercariae were collected from naturally infected sweetfish by peptic digestion; a total of 8 dogs was orally infected with 10,000 metacercariae respectively. Two dogs were killed on 3 days, 9 days, 4 weeks and 10 weeks after the infection. The postures of worms in histological section of small intestine and of whole worms collected from the fixed intestinal mucosa were examined by light and scanning electron microscope. The results were summarized as follows: The recovery rates of worms were 42.6% on 3 days, 55.0% on 9 days, 33.2% on 4 weeks and 9.8% on 10 weeks after the infection respecively. In histological sections of small intesine, most of worms were found at intervillous spaces as ovoid sections of posterior body. However, many worms, especially in 3 day-old worms, revealed protruded anterior body in glandular lumens of crypt. Some sections of anterior body were bifurcated or sacculated. The worms collected from fixed intestinal mucosa under dissecting microscope exhibited a variety of postures. Many worms showed flat shapes with a concavity or curvatures. However, in many worms, the anterior body made a single or multiple protrusions. By SEM observation of protruded anterior body, a longitudinal groove was found. The frequency of worms with protuded anterior body decreased in 9 days, 4 weeks and 10 weeks than in 3 day-old worms. The above findings indicated that the anterior body of juvenile and adult M. yokogawai protruded to lumens of glandular crypt by folding their lateral protions to make a reversible tubelike structure. Frequent multiple protrusions were considered to be made to adapt the microniche of glandular crypts of dog intestine. PMID- 12888663 TI - Studies on intestinal trematodes in Korea XX. Four cases of natural human infection by Echinochasmus japonicus. AB - Four cases of human infection by Echinochasmus japonicus (Trematoda; Echinostomatidae) were proven by the recovery of adult worms after a treatment with praziquantel (10 mg/kg in single dose) and purgation in February 1984 in Korea. The patients were 3 middle-aged men and 1 elderly woman residing in a riverside village of Kangjin-gun ( Tamjin river), Jeonranam-do, an ever-known endemic area of metagonimiasis. The treatment revealed numerous M. yokogawai worms together with 1-43 (total 54 ) specimens of E. japonicus and/or a few number of other kinds of trematodes or cestodes. The patients had gastrointestinal troubles such as indigestion and abdominal discomfort, probably due to these fluke infections. They have been eating some raw flesh of fresh water fishes such as cyprinoid ones or sweetfish caught from the river. This is the first record of human E. japonicus infection in the literature. PMID- 12888664 TI - [Metacercariae of Echinochasmus japonicus encysted in a fresh water fish, Pseudorasbora parva, and their development in experimental mice] AB - The echinostomatid metacercariae encysted in the gill of the fresh water fish, Pseudorasbora parva were identified through obtaining adult worms after experimental infection to mice. In addition, a brief course of worm development and maturation was observed in this experimental host. The results were as follows: The echinostomatid metacercariae were elliptical, golden yellow, 0.073~0.078 mm long and 0.054-0.065 mm wide. Their head portions were characterized by the presence of a head crown armed with collar spines of total 24 in number and interrupted at the mid-dorsal side of the oral sucker. The average rate of worm recovery from 12 mice (on the 1-21th postinfection days) was 19.4 percent and the rate revealed no decrease in accordance with the increase of infection duration. The worms were collected chiefly from the lower part of the small intestine. After the infection, their sexual maturation was attained in 5 days and their growth in size nearly completed in 7 days. The early growth curve of genital organs was S shape while that of nongenital organs was C form. In 5 day old worms, l or 2 eggs were found from their uteri and the stools of mice revealed echinostomatid eggs from the 5-6th postinfection day. The 7 day old adult worms were ovoid in shape, 0.54-0.69 mm long and 0.29-0.34 mm wide, and characterized by a well developed head crown with 24 collar spines and vitelline follicles distributed from the acetabular level down to the posterior end of body. Based on these characters they were identified to be Echinochasmus japonicus Tanabe, 1926. From these results, it is verified that P. parva is one of the second intermediate hosts of E. japonicus in Korea. PMID- 12888665 TI - [Echinostoma hortense metacercariae naturally encysted in Odontobutis obscura interrupta (a freshwater fish) and experimental infection to rats] AB - The metacercariae of an echinostomatid fluke were detected from a freshwater fish, Odontobutis obscura interrupta caught from the Seom river system, Wonseong gun, Kangwon-do, and experimenally infected into rats to obtain the adult worms for the species identification. A total of 32 metacercariae was detected from 10 fishes (27.8 percent) out of 36 O. obscura interrupta caught from the river. The average size of metacercariae was 153.4 x 149.0 micrometer. Also, it was uniquely specific that thickened membrane was formed around the metacercarial cyst wall. The rats orally infected with those metacercariae were sacrificed 20 days after infection to get the adult worms. The mean length and width of the adult worms were 0.75 cm and 0.13 cm, respectively. The number of collar spines on circumoral disk was 26 to 28, and the end group spines at lateral sides of the oral sucker were 4 on each side. The echinostomatid flukes observed in this study were all identified as Echinostoma hortense according to the morphologies of the cirrus sac, ovary, vitellaria and testes of the adult worms and the shape of eggs. On the other hand, O. obscura interrupta is one of the fresh-water fish that are preferably eaten raw by the rural inhabitants. Although no human case of E. hortense infection by eating O. obscura interrupta has been experienced yet, the possibility is strongly suggested in this study. PMID- 12888666 TI - [A case of hyperinfection syndrome with Strongyloides stercoralis] AB - A case of Strongyloides stercoralis infection wss experienced in a 73-year old Korean female patient, was hospitalized with relapse of cholecystitis. The patient developed cough and dyspnea 17 days after the admission. On the 27th hospitalized day, diarrhoea, nausea, vomiting and abdominal pain started. A number of parasitic larvae were incubated at 25 degrees C for 2 days. Typical fork tailed filariform larvae of S. stercoralis (Bavay, 1876) Stiles and Hassall, 1902, were identified after cultivation. There was no improvement of diarrhoea after the medication with mebendazole. After the administration of thiabendazole, however, diarrhoea was stopped. On the 6th day of medication, S. stercoralis larvae were no more detected, and thereafter no larva was observed by repeated stool examinations upto 2 months after chemotherapy. The patient had the history of administration of steroid for articular rheumatism. Therefore this case seems to be a hyperinfection of S. stercoralis due to an autoinfection and to be the first report on the hyperinfected strongyloidiasis in Korea. Related literature was briefly reviewed. PMID- 12888667 TI - Prevalence, worm burden and other epidemiological parameters of Ascaris lumbricoides infection in rural communities in Korea. AB - The epidemiological status of ascariasis was analyzed in 8 rural villages in Korea, through observation of its epidemiological parameters such as prevalence, worm burden and basic reproductive rate. Total 978 inhabitants were subjected to stool examination and recovery of worms after chemotherapy with pyrantel pamoate. The results were as follows: The worm positive rate in each village was 16.5~79.5%, while the egg positive rate was 9~18% lower, 3.3~66.7%. The average worm burden(among all inhabitants) ranged from 0.21 to 8.44 by villages and the frequency of cases with each worm burden showed negative binomial distributions with 'k' values of 0.38-0.54. The prevalence rates (worm) in each village was almost identical with the theoretical ones from Anderson and May's equation; p=1 (1+M(*)/k)**(-k), where 'p' is worm prevalence and 'M(*)' equilibrium average worm burden. The basic reproductive rate 'R' was calculated from 1.03 to 2.11. It is suggested that, although 'R' in lower endemic areas is approaching to the breakpoint of reinfection (R=1), control programs of ascariasis in Korea should be continued until it becomes below the level nationwidely. PMID- 12888668 TI - [Studies on the comparative migration patterns of Ascaris suum larvae between primary and re-infected mice] AB - In the present study, the effect of primary infection to reinfection with Ascaris suum larvae was experimented in mouse model. Mice were challenged with l,000 infective stage eggs of Ascaris suum. The embryonated eggs were directly introduced into stomach of mice. Reinfection was performed at 50 days after the primary infection with same method as primary infection. Mice were sacrificed 3, 5, 7, 10, 15 and 20 days after infection in both groups respectively. Larvae collected from livers and lungs with Baermann's apparatus were enumerated and measured after sacrifice. Sera of mice were also collected at same time. The results of the experiment were as follows: With antigen prepared from coelomic fluid of adult Ascaris suum and sera collected from mice before reinfection, the production of antibody in experimental mice was confirmed by the gel-diffusion technique. In the livers of reinfected mice, the larvae were recovered up to 10 days after challenge, otherwhile in the primary infected mice, the larvae were observed up to 7 days. The maximum number of larvae were observed in the lungs of primary infected mice on 10 days after inoculation. In the lungs of reinfected mice, maximum number of larvae were recovered on 7 days after, only few larvae were recovered on 10 days after reinfection. As regards the growth of the larvae, the third stage larvae, over 500 micrometer in length, appeared in livers at 5 days after reinfection, but it couldn't be found on 7 days and 10 days after challenge. The third stage larvae continuously developed were observed in lungs of mice from 5 days after reinfection. In conclusion, it was found that development of larvae in livers of immune mice were probably repressed by the immune mechanisms being rised in livers and defence mechanism is also acting by interfering with the process of larval penetration into the lung from the liver. PMID- 12888669 TI - [Number of intrauterine eggs in female Enterobius vermicularis by body length] AB - In order to correlate the number of eggs in female Enterobius vermicularis with their body length and to re-evaluate the number of eggs liberated by gravid females, a total of 203 worms were examined. Those females were removed from naturally infected orphans with mebendazole (100 mg) and had been fixed in 10 percent formalin(Cho et al., 1981). The decent ones which were selected under dissecting microscope were unbroken, neatly fixed females without contaminated eggs on their surface. The worms were measured by their body length, softened in 0.1N NaOH solution overnight, and teased by dissecting needles. And their number of shelled eggs was measured in a counting chamber made as described by Denham et al. (1971). The results were summarized as follows: The observed females, 4.10~9.90 mm long, began to have shelled eggs in uterus when body length was 5.50 mm or longer. The percentage of females with eggs in uterus was as follows by range of body length: 25 percent in 5.50~5.99 mm long, 53.3 percent in 6.00~6.49 mm long, 86.7 percent in 6.50~6.99 mm long, 95.2 percent in 7.00~7.49 mm long and 100 percent in 7.50 mm or longer. The mean and standard deviation of egg number were as follows by the length of females: 19+/-50 in 5.50~5.99 mm long, 734+/ 1,597 in 6.00~6.49 mm long, 1,473+/-1,606 in 6.50-6.99 mm long, 1,530+/-2,055 in 7.00-7.49 mm long, 2,567+/-2,046 in 7.50-7.99 mm long, 5,598+/-2,470 in 8.00~8.49 mm long, 9,318+/-2,651 in 8.50~8.99 mm long, 10,678+/-2,892 in 9.00~9.49 mm long and 13,323+/-1,778 in 9.50~9.90 mm long. The numbers of uterine eggs showed greater individual variation when the female lengths were in range of 5.5~8.0 mm. When the female length was longer than 9.0 mm, the egg numbers were over 10,000 in majority, and showed lesser individual variations. Above results suggested that the egg production in female E. vermicularis began in 28-32 days after infection and that in early stages, the egg production varied by individual worms, but in gravid females longer than 9.0 mm at last deposited 10,000 to 16,000 eggs in their uterus with the least individual variations. PMID- 12888670 TI - An acephalic budding Cysticercus (= Racemose cysticercus) found at the abdominal wall of a man. AB - An acephalic budding Cysticercus of 1.2 cm long was removed surgically at the abdominal wall of a Korean man. The worm revealed abnormal buds on the bladder wall and absence of suckers and hooklets in the scolex body. The buds were of two histologic types; branching bud covered with normal tegumentum and with subtegumental cells of normal density, and buds of proliferated subtegumental cells with lacunae formation. On the bases of the morphologic features, it was identified as a racemose cysticercus. This case confirms that its extracranial location is possible. PMID- 12888671 TI - [Electron-microscopic studies on fine structure and enzyme activity in the axenic and conventional strains of Entamoeba histolytica] AB - The metabolism of Entamoeba histolytica would be affected by various environmental factors, and alteration of the environment was known to affect the fine structure of E. histolytica. The present study was designed electronmicroscopically to investigate the ultrastructure and enzyme activities in the axenic and conventional strains of E. histolytica. The trophozoites of axenically cultivated HK-9 strain and conventional YS-27 and YS-49 strains of E. histolytica were collected and fixed with 4 percent paraformaldehyde/0.1 M cacodylate buffer (pH 7.4). After washing them by centrifugation, 1 percent warm agar was added in the sediment. Solidified agar with the trophozoites was cut into 1 mm(3) cubes, and incubated in the various substrates to observe enzyme activities. Then, the specimen was post-fixed with 3 percent glutaraldehyde/0.1 M cacodylate buffer (pH 7.4) and 1 percent osmium tetroxide/0.1 M cacodylate buffer (pH 7.4), dehydrated in ascending ethanol series and embedded in epoxy resin. These were sectioned on an ultramicrotome and observed with a transmission electron microscope. The procedures for the observation of the fine structure were same as the above, except for the incubation in the substrate. The sections were stained with uranyl scetate and lead citrate. For the observation of the surface of the amoebae, scanning electron microscopy was carried out. The results obtained in the present study are summarized as follows: 1. The fuzzy coat around double-layered plasma membrane of E. histolytica was more irregularly and densely distributed in the conventional strains (YS-27, YS-49 strains) than in the axenic strain (HK-9 strain). 2. The endosomes, button bodies and chromatin material were surrounded by a double-layered nuclear membrane having scattered nuclear pores. The paranuclear body, mono- or double-layered vacuoles, vacuolar membrane whorls, rosette-like cylindrical bodies, aggregation of cylindrical bodies and helical bodies were found in the cytoplasm of the amoebae. Helical bodies and glycogen granules were generally abundant, while a few smooth endoplasmic reticula were observed in the cytoplasm. 3. Alkaline phosphatase activity was mainly demonstrated in the plasma membrane, limiting membranes of vacuoles and smooth endoplasmic reticula. ATPase activity was observed in the nucleus, limiting membranes of vacuoles and vacuolar membrane whorls. 4. Acid phosphatase activity was commonly demonstrated in the limiting membranes an contents of vacuoles, lysosome-like organelles, plasma membrane and the button bodies in the nucleus. The activity was more weakly demonstrated in the HK-9 strain than in the other conventional strains of E. histolytica. No peroxidase activity was observed in the amoeba strains employed in the present study. 5. With a scanning electron microscope, no distinct structural differences were observed between the amoeba strains. All the trophozoite forms of the amoebae showed crater-like depressions and rugged features on the outer surface. PMID- 12888672 TI - [Evaluation of Giardia lamblia detection method in stool specimens fixed with SAF solution] AB - The present study has been designed as a basic study on laboratory diagnosis of giardiasis and to demonstrate a more effective method for the detection of Giardia lamblia cyst with the inherent advantages of minimizing both the number of stool examinations required and the interval of stool collections for estimating the real state of prevalence in the shortest time possible. There were 3 subject groups of 75 children each currently residing in an orphanage in Gunsan city, Jeonbuk province from which stool specimens were collected every day, every other day, and every 3 days. The procedure is as follows: Resuspend the fixed sample after fixation with SAF solution. Centrifuge the sediment for 1 min. at 2,000 rpm after straining through gauze into a tube. Divide the sediment into 3 parts and use them for direct fecal smear, formalin-ether concentration (MGL) and zinc sulfate (ZnSO(4)) floatation techniques. The results are summarized as follows: Overall infection rate after 10 trials showed a 60 percent positive indication. The positive rate among children under 4 years old was significantly higher than the rate in children over 4 years old. No significant difference in rate by sex was observed. The results of examinations by direct fecal smear and MGL techniques appeared more accurate than that obtained by ZnSO(4) floatation method as indicated by a higher positive rate. Of all three methods concerned, combinations of two demonstrated a higher positive rate than that shown by any one alone. In three consecutive examinations under varying conditions such as different days, the cyst detection rate by MGL technique indicated 83 percent. In 5 examinations under the same varying conditions, the indicated rate was 94 percent. The interval of stool collection proved to be insignificant for the cyst detection rate. In conclusion, both MGL method and modified fecal direct smear can provide a good cyst detection rate of G. lamblia provided that more than 3 consecutive examinations of stool under varying conditions are carried out. PMID- 12888673 TI - [Protective immunity against Naegleria meningoencephalitis in mice] AB - This study is to verify the protective ability against experimental Naegleria meningoencephalitis by immunization with Naegleria fowleri in mice. Naegleria fowleri, strain 0359, and Naegleria gruberi, strain EGB, were used in this study, and cultured in CGVS medium axenically. Inbred BALB/c mice, weighing about 20g, were immunized by three intraperitoneal injection of 1 x 10(6) N. fowleri trophozoites at the interval of one week. This N. fowleri trophozoites antigen was fixed with 5 percent formaldehyde. N. fowleri trophozoites from culture were homogenized with sonicator at 4 degrees C as monitored by phase contrast microscopy, and their membrane and cell content preparations were made for the immunization of mice. Their inoculation dose in volume was equivalent to the 1 x 10(6) trophozoites in each injection for immunization. And N. gruberi trophozoites, which was fixed with 5 percent formaldehyde, were also used for immunization. Mice were inoculated intranasally with 5 x 10(4) N. fowleri trophozoites in a 5 microliter suspension under anesthesia by as intraperitoneal injection of about l mg secobarbiturate. Nervousness, rotation or sluggish behaviour were observed in the mice which were infected with N. fowleri. Necrotic lesion was demonstrated in the anterior portion of brain, especially in the olfactory lobe. The inflammatory cell infiltration with numerous N. fowleri trophozoites was noticed. This pathological changes were more extensive in the control than in the experimental groups. Mice were dead due to experimental primary amoebic meningoencephalitis that developed between 8 days and 23 days after inoculation. Mortality rate of the mice was low in the immunized experimental group. Mean survival time, which is the survival duration of mice from the infection to death, was prolonged significantly in the immunized mice except in the mice immunized with N. fowleri membrane. Even in the mice immunized with N. gruberi, survival time was delayed. In summary, the effectiveness of immunization is demonstrated in terms of protective immunity against Naegleria meningoencephalitis in mice. PMID- 12888674 TI - [Significance of Toxoplasma antibody titers by indirect latex agglutination tests in pregnant women and pelvic tumor patients] AB - A total of 377 pregnant women, 43 pelvic tumor patients and 80 of multiphysic health center persons as controls were examined by indirect latex agglutination test in order to evaluate Toxoplasma antibody titers at Kang-Nam St. Mary's Hospital in Seoul. Throughout this survey, 1:32 or more titers of diluted sera were regarded as positive. The 337 samples of test sera in pregnant women showed negatives in 319 cases (84.6 percent), 1:2 in 44 cases (11.7 percent), 1:4 in 9 cases (2.4 percent), 1:8 in 2 cases (0.5 percent), 1:16 in 1 case (0.3 percent) and 1:32 in 2 cases (0.5 percent) respectively. The 43 samples of test sera in pelvic tumor patients showed negatives in 29 cases (67.4 percent), 1:2 in 8 cases (18.6 percent), 1:4 in 1 case (2.3 percent), 1:16 in 2 cases (4.7 percent), 1:32 in 1 case (2.3 percent) and 1:128 in 2 cases (4.7 percent). The 80 samples of test sera in multiphysic health center persons as controls negatives in 56 cases (70.0 percent), 1:2 in 19 cases (23.8 percent), 1:4 in 3 cases (3.8 percent), 1:8 in 1 case (1.3 percent) and 1:128 in 1 case (1.3 percent). Among total 420 study cases, 5 cases (1.2 percent) showed positives, and they were 2 cases (0.5 percent) of pregnant women and 3 cases (7.0 percent) of pelvic tumor patients. One case (1.3 percent) out of 80 control sera showed positive result. PMID- 12888675 TI - A description with scanning electron microscopy on the tick Ixodes persulcatus (Schulze, 1930) male and female specimens. AB - The surface fine structures of Ixodes persulcatus (Schulze, 1930) male and female specimens were observed by means of a scanning electron microscope. A brief review on the biology of the ticks and their disease relationships was also presented. The sexual dimorphism of the specimen was marked; the male was quite smaller than the female. The genital groove was well developed and deep, the anal groove was distinct and characteristically extending anteriorly around the anus. The 4th article was much reduced and situated on the top of the 3rd article ventrally. The hypostome dentition was usually 3/3. The bottom of the basis capitulum of the male specimen was strictly straight in shape. This species was regarded as one of the most important vectors for infectious diseases of migrating birds. PMID- 12888676 TI - Scanning electron microscopic observations on the surface structure of the tick Boophilus microplus(Canestrini, 1887) female specimens. AB - Boophilus microplus (Canestrini, 1887) female specimens were subjected to scanning electron microscopy for the observation of surface fine structures. Morphological characteristics observed were summarized as follows: The palpi were wider than long, and compressed, ridged dorsally and laterally. There were usually four pairs of ventro-internal setae on the articles. The hypostome dentition was usually 4/4 but occasionally 5/5. The basis capituli were hexagonal dorsally and the porose area was distinct with the holes developed well. There were numerous folds resembling finger prints around the female genital pore. A pair of anal lobes with specified setae on them was also recognized as one of the morphological characteristics. PMID- 12888677 TI - A Human case of thelaziasis in Korea. AB - A case of thelaziasis in 36 year-old Korean woman who suffered from foreign body sensation of both eyes and itching of conjuctiva, was reported in August 1984. Four thread-like white worms was removed from cojunctival fornix of both eyes. This is the 12th case of thelaziasis in Korea(summarized from text). PMID- 12888679 TI - A record of Diplozoon nipponicum Goto, 1891 found from Cyprinus carpio nudus in Korea. AB - Diplozoon nipponicum, a monogenean trematode from Cyprinus carpio was described according to the characteristic findings of X-form adult, metrical data, and presence of a pair of sticky glands. It is the first record of the presence of Diplozoon nipponicum in Korea. PMID- 12888678 TI - Two cases of imported cutaneous leishmaniasis in Korea. AB - Two Korean men who had worked in Saudi Arabia complained of skin ulcers in the right wrist for 5 months or in the face and left forearm for 4 months each. Ulcers accompanying crust was observed in both cases. The characteristic cellular destruction and amastigote in the cytoplasm of histiocyte supported the diagnosis of cutaneous leishmaniasis. Those are 17th and 18th imported cases of cutaneous leishmaniasis in Korea(summarized from text). PMID- 12888680 TI - [Rhabditis sp. infected cases in rural school children] AB - Five cases infected with Rhabditis sp. were detected in a survey to examine the stool specimen from rural primary school children. A large number of the larvae of Rhabditis sp. detected by the direct cellophane thick smear were cultured by the filter paper method. The examination was carried out in April through June 1980 in Tangjeong-Myon, Ahsan-Gun, and in September 1983 in Sandong-Eup, Yeongwol Gun. The results obtained in this study were as follows: Prevalence: Out of 925 children, 5(0.54%) children were found to be positive. The number of the detected larvae was 110/0.1 gm of feces in case 1, 35 in case 2, 130 in case 3, 86 in case 4 and 62 in case 5. Larvae: The larvae were prepared by means of the direct fecal smear and measured by a micrometer equipped in the microscope. Twelve(12) through 15-day old larvae in culture were 197.1 micrometer long in average, and the maximum size of the matured stage larvae was 884.0 x 25.9 micrometer. However, the length variation was ranged as 173.0 to 884.0 micrometer. Adults: The size of clubbed adult female was 1,357 micrometer(1,176~1,419) in length and 80 micrometer(79~82) in width. Length of buccal cavity was 33 micrometer. A long cylindrical esophagus (273 micrometer) of the worms with a valved posterior cardiac bulb and with median bulbar swelling was morphologically indicated. Distance from mouth to vulva was occupied 58 percent of body length. Male worm was 1,006 micrometer (890~1,148) in length and 49 micrometer(48~49) wide. Caudal alae of bursa and spicules (75 micrometer in length) were well developed. Eggs: The oval shaped eggs in the female uterus, when cultured, were 66 x 56 micrometer in size, and the eggs laid by the adult could not be detected. So, reproduction might be through to be ovoviviparity. The five cases were re-examined during the period from the 1st to the 3rd week after stool examinations, but Rhabditis sp. were detected again in 4 cases in 1st week. When they were examined in 3rd week, larvae could not be detected. So, it was thought that the infection of Rhabditis sp. to humans was facultative. PMID- 12888681 TI - Effect of anthelmintics on the early stage of Enterobius vermicularis. AB - In order to determine the susceptible age of Enterobius vermicularis to anthelmintics and to observe the chronologic growth of female E. vermicularis in man, experimental infections were done. About 500 eggs were challenged to 19 volunteers. After 4, 8, 16, 20, 24, 28, 32 and 35 days of infection, each case was treated by either mebendazole or pyrantel pamoate. On the 40th day of infection all cases including control were treated again to terminate the expermental infection and to evaluate the effect of previous treatment. Each case collected 3-day stools to harvest the expelled worms. The results could be summarized as follows: The infection rates of females were in range of 0.6~13.1% in control cases. Because the collected worms showed comparable growth and development by day, the worns were concluded to be derived from experimental infection. Cases that were treated with mebendazole on 4, 8 and 16 days after infection expelled 37.5%, 2.5% and 67.5% of the number expelled by a control case on the 40th day. Cases treated thereafter expelled no worms on the 40 days. Cases that were treated with pyrantel pamoates on 4, 8, 16, 24, 28, 32 and 35 days, expelled 90.7%, 25%, 45.3%, 8%, 2.7%, 5% and 29.3% of the number collected from control cases in respect. All the worms collected were females. The total body length increased consistently and comparably from the 20th day of infection. Those collected on the 20th day were 2.5~3.0 mm long with vigina, sac-like structure and strands of ovaries; 24 day-old worms may have short uterus, 28 day old worms had long uterus without eggs, 32 day-old worms began to produce eggs, 35 day-old worms showed wide variations in egg deposit in uterus, and 40 day-old worms had uterus filled with eggs from vulva to anal levels. From the above results, it was inferred that the life span of female Enterobius vermicularis was longer than 40 days, and the developmental stages of worms younger than 16 days resisted considerably to both mebendazole and pyrantel pamoate. PMID- 12888682 TI - Studies on intestinal tematodes in Korea: XVI. Infection status of loaches with the metacercariae of Echinostoma hortense. AB - The infection status of the loaches, Misgurnus anguillicaudatus, with the metacercariae of Echinostoma hortense, was studied in Korea. A total of 154 loaches purchased at 4 local makets (Seoul, Kimhae, Naju-gun and Kangjin-gun) were examined their infection rate as well as the density and location of the metacercariae in the fish body. The results are as follews: The loaches carrying the metecercariae of E. hortense were 64 (41.6%) in total number and the metacercarial density ranged 1-29 per infected loach with an average value of 8.1. The highest infection rate and metacercarial density were obtained from the loaches purchased at Kangjin-gun, Jeonranam-do. The metacercaria of E. hortense were chiefly distributed in the distal intestinal wall and the adjacent mesentery, the perianal tissues, and the head and gill of the loaches examined. From the results, it is concluded that the loach is one of the important second intermediate hosts of E. hortense in Korea, and their infection rate and metacercarial density are considerably high. PMID- 12888683 TI - Studies on intestinal trematodes in Korea: XVII. Development of egg lying capacity of Echinostoma hortense in albino rats and human experimental infection. AB - The worm development and egg laying pattern of Echinostoma hortense(Trematoda: Echinostomatidae) were studied in albino rats and the brief clinical course was observed in human volunteers. A total of 21 rats were infected with 20~69 metecercariae each and two humans were with 7 and 27 metacercariae, which were collected from loaches. For recovery of worms, the rats were sacrificed at irregular intervals from the 6th to 150th day after infection and the human volunteers were treated with praziquantel and purged with magnesium salt on the 26~27th day. The stools of the rats and humans were examined for the eggs. The results were as follows: The worm recovery rate from the rats was not affected by the increase of infection time but varied individually; 9.1~50.0% (31.1% in average). From humans, 14.3% and 37.0% (32.4% in average) of challenged were recovered. In the rats, it was revealed that the worms rapidly grew for the first 14 days to become 7.59 mm in average length and 1.17 mm in average width but the growth became much slower thereafer until the 150th day; 7.95 mm in length on the 21th day, 9.04 mm on the 28th day, 10.21 mm on the 49th day and 12.62 mm on the 150th day. During the early stage of infection, the growth of genital organs(male or female) was expressed as sigmoid curves whereas non-genital organs(such as suckers) was simply as straight lines. The prepatent period of this fluke was 10~12 days in the rats and 16~17 days in men. After the start of oviposition, the egg production by the worms remarkably increased, reached maximum on the 32~33th day, followed by decrease thereafter. The maximum value of E.P.G./worm was 390. The major subjective symptoms in human volunteers were abdominal pain and diarrhea during the early stage of infection. The results show that human is as susceptible as the rats to E. hortense infection and the amount of egg production in the rats is greatly affected by the age of worms. PMID- 12888684 TI - [Two cases of natural human infection by Echinostoma hortense and its second intermediate host in Wonju area] AB - Two cases of human Echinostoma hortense infection and their probable infection source were identified by praziqantel (Distocide) treatment of the patients and by examining two kinds of fresh water fish which were eaten raw by them. The result of the research can be summarized as follows: The patients, each aged 31 and 30, were residing in the same house in Wonju City, Kangwon Province. The first case was hospitalized due to epidemic hemorrhagic fever (E.H. fever) and the second case was healthy but had slight degree of abdominal pain and diarrhea from time to time. In the stool examination, eggs of E. hortense (114.3 x 71.0 micrometer average from the first case and 119.1 x 68.3 micrometer average from the second) were found. By administering single dose of praziquantel (10~15 mg/kg) and purgation with magnesium salt to them, six adults of E. hortense were collected from the diarrheal stools of the second case. By examining 84 Moroco oxycephalus and 20 Carassius carassius which were captured at the place where the two patients had captured and eaten the fresh water fish, the metacercariae of Echinostoma sp. were found from 3 (3.5 percent) M. oxycephalus. After the experimental infection of 3 isolated metacercariae to one albino rat three adults of E. hortense were recovered. By the present study, the two patients revealing the echinostomatid eggs in their stools were proven to be infected with E. hortense and to be the second and third human cases of this fluke infection in Korea. Moroco oxycephalus harboured the metacercariae of E. hortense and appeared to be a new second intermediate host. PMID- 12888685 TI - Studies on intestinal trematodes in Korea: XVIII. Effects of praziquantel in the treatment of Fibricola seoulensis infection in albino rats. AB - The therapeutic effect of praziquantel on Fibricola seoulensis infection were studied by experimental infection of albino rats with the metacercariae obtained from the snakes. Total 51 albino rats were infected each with 500 or 1,000 metacercariae in number through intragastric tubes. One or four weeks later the rats were treated with 1, 5, 10 or 20 mg/kg sigle dose of praziquantel and sacrificed after 3 days or 10-120 minutes to search for the worms in their small intestine. The worms recovery rate at 3 days after the treatment was 10.7% in 1 mg/kg does group and 0.03~0.2% in 5, 10 and 20 mg/kg group, while that of untreated control was 51.3%. The minimum effective dose to treat F. seoulensis infection in the rats is considered to be 5 mg/kg in sigle dose. By observing the distribution pattern of worms in the small intestine after the treatment, dislodgment of the worms froms the duodenum, thier normal habitat, to the lower portion was recognizable within as early as 10 minutes. The majority of worms was found to have descended to the ileo-caecal portion after 60-120 minutes. PMID- 12888686 TI - Studies on intestinal trematodes in Korea XIX. Light and scanning electron microscopy of Fibricola seoulensis collected from albino rats treated with praziquantel. AB - An experimental study was performed to observe the in vivo effects of praziquantel on the light and scanning electron microscopic morphology of Fibricola seoulensis. The metacercariae were obtained from the snakes and 1,000 in each number was orally given to total 15 albino rats; 5 controls and 10 treatment group. Seven days later the 10 rats were treated with 10 mg/kg praziquantel and sacrificed 1~24 hours later to search for the worms from their small intestines. The major light microscopic changes in the drug-exposed worms were early contraction followed by relaxation of especially their forebody, vacuolization of the tegument and subtegumental parechymal layers, and narrowing of the intestinal lumens. The scanning electron microscopic findings were characterized by formation of numerous blebs followed by rupture and subsequent destruction of their whole tegument. These results show that the change in worm body is not confined to the tegument but extends to deeper parechymal portions and also occurs in their intestines. It is suggested that the drug uptake by the worms should be either through their tegument or through the digestive tract. PMID- 12888687 TI - A histopathological study on the intestine of mice and rats experimentally infected by Fibricola Seoulensis. AB - A histopathological study was carried out on the duodenum of mice and rats experimentally infected by F. seoulensis. Each mouse was infected wit 500 metacercariae and killed after 1, 2, 3 days, 1 and 2 weeks from infection. Each rat was given 1,000 metacercariae and was examined after 1, 2, and 4 weeks from infection. The duodenal tissue sections of mice and rats were stained with hematoxylin and eosin, and PAS stained for the rats of 1 week group. The pathological findings are summarized as below. There were no differences in mucosal findings between the mice and the rats, adn between the location of duodenum, 1 and 5 cm distal to the pylorus. Each worm embraced a villus exclusively with its foliate forebody which was inserted into the intervillous spaces. The fluke pinched villous epithelia using its oral and ventral suckers. The tribocytic organ destroyed the villous epithelia deeply up to the stroma after 3 days from infection. Apparent villous changes were observed in the mice after 3 days from infection. Villous changes were shortening, widening, blunting or fusion. The villous stroma showed edema, microscopic hemorrhage, capillary congestion, dilatation of lymphatics and inflammatory cell infiltration. The cells were lymphocytes, plasma cells, eosinophils and giant cells. Rarely submucosal and transmural inflammation was encountered. PMID- 12888688 TI - [A study on the infection status of tadpoles and frogs by the metacercariae of Fibricola seoulensis in Korea] AB - A total of 220 tadpoles was captured in 6 areas and total 117 frogs, Rana nigromaculata, were collected in 11 areas in Korea. They were examined for their infecion status by the metacercariae of Fibricola seoulensis by peptic digestion technique and by histological observation with hematoxylin-eosin staining. This study was carried out from August 1983 to September 1984. Followings are the results. The tadpoles of R. nigromaculata were positive for the metacercariae from 3.3 percent to 100 percent by area. The number of metacercariae per infected tadpole ranged from 1 to 584, and the mean number per tadpole ranged from 7.6 to 221 by area. The metacercariae from 16 tadpoles were counted by the body portion. A great majority of the metacercariae was collected from abdominal cavity, 98.3 percent of 484 counted larvae. And 6(1.2 percent) larvae were from proximal tail and 2(0.4 percent) from trunk. Histological sections of tadpoles showed many metacercariae in abdominal cavity but none in other parts. The larvae were free in the spaces among intestinal loops or around primitive liver. A few larvae were in duct-like tissues near trunk wall. There was little infiltration of inflammatory cells. The metacercarial infection rates of frogs ranged from 0 percent to 100 percent by area. The larval burden was 1 to 470 by infected frogs, and mean number ranged from 1 to 175.6 by area. By above results, it is suggested that the cercariae of F. seoulensis may infect R. nigromaculata already in the stage of tadpole. Almost all of the metacercariae were concentrated in abdominal cavity of tadpoles. According to the infection status of frogs, this fluke is prevalent almost nation-widely in rice paddies in Korea. PMID- 12888689 TI - [Experimental studies on the second intermediate hosts of Clonorchis sinensis IV. Observations on the fate of metacercariae of Clonorchis sinensis in the fish host, Cultriculus eigenmanni] AB - In order to judge appropriately the suitability as the second intermediate host of Clonorchis sinensis, the present experiments were proposed to survey the metacercaria of C. sinensis in the fish host, Cultriculus eigenmanni, in the field, and to observe, by experimental infection of the fish with Clonorchis cercaria, the penetrating ability of the cercaria, maturity, the process of degeneration and extinction, and infectivity of the metacercaria. The following is a brief summary of the leading facts gained through the experiments. All the ten fish (mean body weight 13.4 +/- 3.43g) which were caught in Ok-ku irrigation reservoir were infected with the metacercariae and the average number of the metacercariae found in the fish body/gram was 4.55. The fish was easily subject to invasion of the cercaria, and the examination of the cercariae after having the fish infected disclosed that 24 hours after the infection, all of the cercariae formed their cysts in muscle and the metacercariae kept growing, that 7 days later the metacercariae were found folding their bodies twice, and that in 15 days the characteristic structure of the metacercariae was complete and made a vigorous rotary movement intermittently. In the case of control fish, Pseudorasbora parva, both penetrating ability of the cercariae and the infective degree of the metacercariae to control host were stronger as compared with the fish host because that the control fish had higher suceptibility to C. sinensis. Afterwards, the metacercariae in the fish host came to a state of maturity and beyond this stage some metacercariae started the process of degeneration and extinction in as early as 30 days, and as more days elapsed, mortality of the metacercariae was not uniformly increased but difference of mortality appeared to be according to the individual of fish host. That is, in 390, 540 and 586 days, all of the metacercariae in the fish hosts became dead, meanwhile almost all of those found in the fish host of 596 days after the infection survived (mortality; 8.51 percent). Percentage of orally administered metacercariae developing to maturity in rabbit is 10.46 percent in the last case. As the days were progressed the shapes of the metacercariae in the fish host tended to change from ellipsoid to almost round shapes, the length became shorter while the width was wider. The results shown above revealed that C. eigenmanni was, though it could not be proper host than P. parva, suitable as the second intermediate host in addition to our previous observation results. PMID- 12888690 TI - Seasonal variations of metacercarial density of Clonorchis sinensis in fish intermediate host, Pseudorasbora parva. AB - The seasonal variations of the rate and intensity of metacercarial infection of C. sinensis in P. parva were observed. The fish were collected at Sun-Am river which located in Kim-Hae City, Kyong-Sang-Nam Do(=Province), Korea, from March 1983 to February 1984 every month. A total of 788 fish was examined. The number of metacercariae in each fish was individually counted after the individual digestion by artificial gastric juice. The result was as follows: During one year, 513(65.1%) out of 788 fish were infected with metacercariae. In May, June, July and September, the infection rates ranged from 82. 0 % to 98. 6% whereas the rates was relatively low in March, April, November and February raning from 11. 4% to 64.7%. The intensity of infection was similar with those of infection rates. The mean intensity per infected fish was 103.0 and standard deviation was 118.9 throughout one year. The highest mean intenstiy was in June(294. 8) and the lowest in Novebmver(11.1). The observed frequency of fish with certain intensities of metcercariae were fitted to theoretical equations derived from negative binomial distribution in March, April, November and February(p > 0.05). Meanwhile, the equation of lognormal distribution were fitted with the observed frequencies in May, June, July and September(p > 0.05, p > 0.75). The variance/mean ratio varied by month. The value was the highest in July(814.3) and the lowest in November(158.8). Unlike our hypothesis, the metacercarial density of Clonorchis sinensis in its the most favourable fish host, Pseudorasbora parva showed considerable seasonal variations in the hyperendemic area. The possible factors were discussed. PMID- 12888691 TI - [Application of micro-ELISA in serodiagnosis of fascioliasis in cattle] AB - Fascioliasis in cattle is one of the most common and very serious trematode diseases in Korea. In the present study, the enzyme linked immunosorbent assay (ELISA) was applied in the diagnosis of fascioliasis using antigen of Fasciola hepatica, peroxidase of conjugate anti-cattle IgG and orthophenylenediamine as a substrate by micro-method technique of Voller et al. (1976b) and MacLaren (1978) with a slight modification. Results obtained from the present study are as follows: In assay for optimal dilution of stock antigen, the antigen (protein contents; 0.8 mg/ml) was diluted from 1/50 to 1/600 with carbonate buffer (pH 9.6), and then absorbance values were measured with 1/100 diluted sera. The regression equations between the OD values of ELISA and dilution of antigen were log Y=-0.181-0.00127X in infected sera, and log Y= -0.578- 0.000879X in normal sera. The significantly higher (p<0.05) OD value was observed in the former. In assay for optimal dilution of sera, the sera were diluted from 1/25 to 1/400 with in PBS/ Tween 20(pH 7.4), and absorbance values were measured with 1/200 diluted antigen. The regression equation between the OD values of ELISA and dilution of sera were log Y=-0.1540-0.0007238X in infected sera and log Y=-0.4834-0.00116X in normal sera. The former was higher than the latter (p<0.05). In the 27 cases of negative intradermal test, OD values of the ELISA are 0.447 +/- 0.144, the 95 percent confidence interval (Mean + 2 x SD) of the values was 0.735, and there was no case over the values. Therefore, the sensitivity of the antigen to diagnose fascioliasis was 100 percent in the negative case. The OD value 0.7 which is designed as a criterion (detection level of positive one) is useful for the performance of the ELISA in fascioliasis. According to the OD value of criterion in the regression equations, the optimal dilutions of stock antigen and serum were 1/250 and 1/100, respectively. In the 58 cases of fascioliasis from which the adult could be found in the bile ducts, the OD value was 0.846 +/- 0.224). The 75 percent (44 cattle) among them had higher value with compared to the criterion, and the 60 percent (20 cattle) of the cases of proliferative cholangitis of 33 cattle which had been infected previousely with Fasciola sp. is higher than the criterion. Prevalence of fascioliasis was 43.4 percent in the application of the ELISA to 272 cattle which were reared in Jeonbug district. PMID- 12888692 TI - Epidemiological survey of Paragonimus westermani in Ulchin County, Kyungpook Province, Korea. AB - In order to determine the epidemiological pattern of the Paragonimus westermani, the infestation rates of the cercarial and metacercarial larvae of digenetic trematodes in the snail and crayfish host, and the prevalence of Paragonimus westermani among the residents in the vicinity of the stream Namdae, Wyangpi, and Kwang, in the Ulchin county, Kyungpook province were studied from March to October in 1984. The population density of the snails per square meter of the habitats ranged form 5 to 25, with average of 15. Among the seven habitats, one, Ducheon, had snails infested with the cercariae of Paragonimus westermani, and the proportion of infested snails was very low, the average being 0.152 per thousand. Of six hundred and four crayfish examined, 113 or 18.7 per cent harboured the encysted larvae of Paragonimus westermani. A high infestation rate for the metacercariae in the crayfish was found in two habitats; 39.0 per cent in the Ducheon and 21.6 per cent in the Sokwang. The prevalence of Paragonimus westermani among the residents in Ulchin county was relatively high, 25.8 per cent by the Paragonimus intradermal tests and the difference in the rate of infection between males and females was found to be significant(t>2). The results obtained in this study indicate that endemic foci of Paragonimus westermani exist in Ulchin county and the prevalence of this lung fluke among the residents is relatviely high. PMID- 12888693 TI - [Ultrastructure of the integument of adult Paragonimus westermani] AB - The present study was performed to observe the ultrastructure of the integument of adult Paragonimus westermani. Dogs experimentally infected with 60 metacercariae of P. westermani were autopsied 4 months after the infection. Adult P. westermani were extracted from the dogs and the fine structure was studied by means of scanning and transmission electron microscope. The findings are as follows: Scanning electron microscopic findings showed that the spines and the papillae are distributed at whole body surface but the well developed spines or papaillae are shown around the oral suckrer and ventral sucker. At the end of the body, excretory pore was found, the shape was irregular. Transmission electron microscopic findings showed that plasma membrane, tegument, basal lamina, connective tissue, circular muscle layer, longitudinal muscle layer, nerve axon and tegumental cell were observed. In higher magnification, plasma membrane and bar-shaped granules were found at the outer surface of the tegument. PMID- 12888694 TI - [Studies on the intestinal fluke, Metagonimus yokogawai Katsurada, 1912 in Korea Geographical distribution of sweetfish and infection status with Metagonimus metacercaria in south-eastern area of Korea] AB - The present survey was undertaken to observe the geographical distribution of metacercaria among the sweetfish, Plecoglossus altivelis, the most important second intermediate host of Metagonimus yokogawai, which were collected during the period from 1983 to 1984 in south-eastern coastal areas of Korea. The results obtained were summarized as follows: 1. A total of 668 sweetfish collected from 70 chons (=streams) in the above coasta1 areas was examined for the metacercariae, and 432(64.7 percent) among them were infected with metacercariae of M. yokogawai. The mean number of metacercariae in a fish was in the range from 0 to 29,604 and the mean number of the metacercariae in a fish was 640.3. 2. It was observed that the mean infection rate showed 46.4 percent in Gangweon-Do (=province). The infection rates varied from stream to stream showing 100 percent in Namdae stream(Gangnun-city), Obsib-stream(Samchok-gun) and Dongmak, 95.0 percent in Hosan stream, 90.0 percent in Gungchon stream, and Namdae stream (Yangyang-up), 80.0 percent in Hwasang stream, 50.0 percent in Daebong stream, 45.0 percent in Yonghwa stream, 30.0 percent in Sangchon stream and Sachon stream, 20.0 percent in Munam stream and Okgyoe stream, 15.0 percent in Imweon stream, 10.0 percent in Namchon stream. And no metacercarial infections in their fish host were found in the areas of Mulchi stream, Sangun stream, Gosamun stream, Jonchon stream snd Gagok stream. 3. It was observed that the mean infection rate showed 50.9 percent in Kyongsangbuk-Do(=province). The infection rates also varied from stream to stream showing 100 percent in Songchon stream, Osib stream (Yongdok-gun) and Daejong stream, 60.0 percent in Pyonghae stream snd Gunmu stream, 50.0 percent in Buhwung stream, 25.0 percent in Nagok stream, 20.0 percent in Oangbi stream, 10.0 percent in Namhodong stream. And no metacercarial infections in their fish host were found in the areas of Namdae stream (Uljinup) and Choksan stream. 4. It was observed that the mean infection rate showed 67.6 percent in Kyongsangnam-Do(=province). The infection rates varied from stream to stream showing 100 percent in Taehwa river, Hwiya river, Inchon stream, Miryang river, Nam river, Hwagae stream, Koha stream, Sosang stream, and Tongchon stream, 90.9 percent in Yoncho stream, 90.0 percent in Hoihwa stream, 80.0 percent in Chindong stream, 66.7 percent in Chingyo stream, 40.0 percent in Hoyam river. And no metacercarial infections in their fish host were found in the areas of Chinjon stream, Sanchon stream and Jukchon stream. 5. It was observed that the mean infection rate showed 94.6 percent in Chollanam-Do(=province). The infection rates varied from stream to stream showing 100 percent in Dongchon stream, Isachon stream, Somjin river (Goksong-gun), Somjin river (Kurye-gun), Sosi stream, Gokchon stream, Kohung stream, Kwansan stream, Youi stream, Unjon stream and Apnae stream, 92.3 percent in Tamjin river, 90.9 percent in Okkok stream, 84.6 percent in Songgun stream. And no metacercarial infections in their fish host were found in the area of Yongok stream. 6. On the other hand, the infection rate showed 100 percent in Inchon river of Chollabuk-Do (=province) and Kangon stream of Cheju-Do (=province). PMID- 12888695 TI - Seasonal prevalence of the vector mosquitoes of Japanese encephalitis virus in Kyungpook Province, Korea. AB - In order to determine the seasonal prevalence and population density of vector mosquitoes in Kyungpook province, a survey based on average number of female mosquito per trap-night, were carried out during the period from May to November in 1984. Among the 34,571 mosquitoes collected in Kyungsan county in animal shelters and human dwellings by light traps, approximately 45.0 % were Culex tritaeniorhynchus, 34.0 % per cent Culex pipiens pallens, and 19.0 per cent Anopheles sinensis. By comparison, distribution of mosquito species in Ankang town, in which Japanese encephalitis had been reported in the past, were quite different and A. sinensis was the most abundant species, being constituted in 75.8 per cent, followed by C. tritaeniorhynchus(23. 2 percent), and C. pipiens pallens(0. 6 per cent). In the general patten of seasonal prevalence, C. tritaeniorhynchus first appeared in mid-June, and trapped in large numbers during the periods from mid-August to early September, showing a simple sharply pointed one-peaked curve, while C. pipiens pallens was found to be active through almost the entire season showing irregular curves with several peaks. The trend of seasonal distribution of A. sinensis was similar to that of C. tritaeniorhynchus, but the earliest dates of appearance and disappearance different form that of C. tritaeniorhynchus. The results of hourly catches of mosquitoes by human baited traps have clearly shown the differences in the biting rhythm among the vector mosquitoes of this province, such as the facts that C. tritaeniorhynchus and A. sinensis are rather constantly active all through a night with more or less inconspicuous peaks twice a night, while C. pipiens pallens exhibits an irregular curve, and the numbers of A. vexans nipponii, C. vagans and Armigeres subalbatus are very small and are not sufficient for estimating the biting rhythm. Four genera and fourteen species of larval mosquitoes collected from eighteen kinds of habitats which classified according to the categories followed by Bates(1949). The main breeding sites contributing to peak C. tritaeniorhynchus adult densities in this surveyed areas were evidently ricefields, ground pools, puddles and swamps and marshes, etc. Summarizing the results, this study indicated that the highest population density of C. tritaeniorhynchus was encountered in late August and early September, showing simple sharply pointed one-peaked curve. PMID- 12888696 TI - [Failure of immunization with Naegleria fowleri in mice born to immune mothers] AB - Female BALB/c mice weighing 18-20 g were immunized by three injections of 1 x 10(6) Naegleria fowleri trophozoites intraperitoneally at the interval of one week 6 times for the pregnant mice and 3 times for the offspring mice. One week after immunization the mice were challenged intranasally with N. fowleri trophozoites 5 x 10(4) under secobarbital anesthesia. Experimental primary amoebic meningoencephalitis developed between day 7 and 16 after infection. All mice were dead due to amoebic meningoencephalitis in all experimental groups except in the offspring born to non-immune mothers. Mean of survival time, which is the duration of survival of mice from infection to death, was delayed in the groups of mice born to immune mothers, immune mice born to immune mothers. Active or passive protective immunity against N. fowleri infection was demonstrated in the immunized mice and mice born to immune mothers. But the effectiveness of immunization was greatly impaired in terms of mortality in the immune mice born to immune mothers when N. fowleri was infected intranasally. PMID- 12888697 TI - [Effect of splenectomy on development of primary amoebic meningoencephalitis] AB - To elucidate the effect of splenectomy on the development of experimental primary amoebic meningoencephalitis in mice, the death rate and survival time of mice infected intranasally with Naegleria fowleri trophozoites 5 x 10(4) cultivated in CGVS medium were compared according to the age when splenectomy was done, and post-operation until experimental infection. Immunodiffusion was undergone to detect the presence of serum antibody due to N. fowleri infection in mice. Polyacrylamide gel electrophoresis was done to compare the protein fractions of mouse serum in each experimental groups. In experiment I, splenectomy was done 3 weeks and infection 4 weeks after birth, the death rate of control, sham operated and splenectomized group were 100 percent, 85 percent and 95 percent, and the mean survival time after infection 7.3 days, 7.5 days and 7.8 days, respectively. In experiment II, splenectomy was undergone 3 weeks and infection 6 weeks after birth, the death rate of control, sham operated and splenectomized group were 95 percent, 95 percent and 95 percent, and the mean survival time after infection 12.1 days, 11.5 days and 11.5 days, respectively. In experiment III, splenectomy was done 5 weeks and infection 6 weeks after birth, the death rate of control, sham operated and splenectomized group were 95 percent, 90 percent and 95 percent, and the mean survival time after infection 8.1 days, 8.3 days and 8.5 days, respectively. By Ouchterlony immunodiffusion, anti-N. fowleri antibody in the serum of mouse with primary amoebic meningoencephalitis was detected against a N. fowleri antigen, which was prepared by ultrasonication of N. fowleri trophozoites, each reacting two lines of precipitation. The patterns of serum fractions by polyacrylamide gel electrophoresis were different between control and sham operated groups from splenectomized group in fraction II, III and V, the sera of which were collected after N. fowleri infection. This results may be summarized as that splenectomy has no effect on the development of primary amoebic meningoencephalitis in mice. PMID- 12888698 TI - [Effect of prednisolone treatment on the experimental inducement of primary amoebic meningoencephalitis] AB - Present study aimed to elucidate the immunosuppressive effect of prednisolone on Naegleria fowleri infection in mice. N. fowleri was cultured in CGVS medium (Willert and Le Ray, 1973). White female mice, weighing about 18 g, used for experiments were divided into five groups; untreated control group, prednisolone treated groups (before, during and after infection), and only prednisolone treated group. In the prednisolone treated group, the hormone was injected intramuscularly 5 doses of 10 mg/kg every other day. According to designated time of treatment, each mouse was challenged with 1 x 10(5) N. fowleri intranasally. Changes of body weights, clinical manifestations and number of dead mouse were observed. Brain and lung tissues of dead mice were cultured in the non-nutrient agar (Kasprzak and Mazur, 1972), or stained with hematoxylin-eosin for the examination of histopathological changes. Results of the experiment are summarized as follows: Mortality among the prednisolone treated groups was higher than that in untreated control group, and among the treated groups, the pretreated group showed shorter survival time. Body weights among untreated control mice showed no significant increase, however, treated groups of mice showed the decrease during the administration and recovery of the weights were observed at 2 to 3 days after the completion of treatment. In the treated control groups, the infected mice began to show the pathologic findings 5 days after infection while the untreated mice began to show the findings 8 days after infection. Tissue damages in brain and lung occurred due to virulence of amoeba were more severe among treated mice than that in untreated control group. The above mentioned results suggest that the treatment with prednisolone weaken the resistance of mice against N. fowleri infection, and probably induce more severe primary amoebic meningoencephalitis. Especially severe pathological findings were shown in pre-treated group, compared with untreated group. PMID- 12888699 TI - Plecoglossus altivelis as a new fish intermediate host of Heterophyopsis continua. AB - P. altivelis from Korea republic is recorded as a new host for H. continua which is described and figured. PMID- 12888700 TI - A preliminary observation on watery content of small intestine in Metagonimus yokogawai infected dog. AB - The intestinal contents of a dog infected with 49,732 M. yokogawai for 8 weeks were normal but in a 4-week-old infection the intestinal water content correlated roughly with the number of worms. Most worms were found 40 to 102 cm along the small intestine but the entire length was parasitized. The concentrations of Na+, K+ and Cl- decreased and the concentration of HCO3- and pH increased along the small intestine while the osmolarity remained roughly constant (324 to 340 mOsm/kg). PMID- 12888701 TI - Drug therapy in the elderly. PMID- 12888702 TI - Short-term efficacy and safety of extended-release fluvastatin in a large cohort of elderly patients. AB - The efficacy and safety of lipid-lowering agents in elderly individuals have not been extensively assessed. This population generally takes more drugs concurrently than middle-aged patients, and are therefore at higher risk of drug drug interactions. This large-scale, randomized, double-blind, placebo-controlled study investigated the efficacy and safety of extended-release (XL) fluvastatin 80 mg once daily for up to 1 year in elderly patients with primary hypercholesterolemia. A total of 1229 patients (mean age, 75.5 years) were randomized. After 2 months of treatment, fluvastatin XL 80 mg significantly decreased plasma lipid levels from baseline compared with placebo; fluvastatin reduced total cholesterol by 25% compared with a decrease of 2.5% in the placebo group, low-density lipoprotein cholesterol was -33% vs. -2.5%, respectively, and triglycerides were -13.3% vs. 2.9%, respectively (p<0.00001). The safety profile of fluvastatin XL was similar to that of placebo. Fluvastatin XL 80 mg once daily was well tolerated and effectively managed plasma lipid profiles in a large cohort of elderly patients. These findings are consistent with data obtained previously in younger recipients of fluvastatin XL 80 mg, and reinforce the safety of fluvastatin in a population at high risk of drug-drug interactions. PMID- 12888704 TI - Drug therapy of heart failure in the elderly. AB - Heart failure is a disorder that predominantly affects older adults, with more than 50% of heart failure hospitalizations occurring in persons over 75 years of age. Unfortunately, most of the major heart failure clinical trials have targeted middle-aged patients with systolic heart failure, and the applicability of these studies to elderly patients, particularly those with preserved left ventricular systolic function, remains uncertain. In this paper, current data on the pharmacotherapy of heart failure in older adults are reviewed, recommended approaches to managing systolic and diastolic heart failure are outlined, and the importance of preventive measures is emphasized. PMID- 12888705 TI - Anticoagulation in the elderly. AB - This review will address the general approach to the management of the typical elderly patient requiring anticoagulation. Most of the data has been derived from studies of patients with nonvalvular atrial fibrillation. Data from postmyocardial infarction trials have also been included. A practical clinical approach to anticoagulation in the elderly is described. Emphasis has been placed on maximizing the benefit and reducing the risk of anticoagulation in the rapidly expanding group of elderly patients aged >/=75 who are at the greatest risk of stroke and are likely to benefit the most from antithrombotic therapy. PMID- 12888706 TI - Thrombolysis in elderly patients with acute myocardial infarction. AB - The efficacy of thrombolytic therapy in the elderly remains a topic of ongoing debate. Although elderly patients account for a disproportionate amount of cardiovascular mortality, they have typically been underrepresented in randomized clinical trials. A meta-analysis of these trials suggests a survival benefit, albeit small, of thrombolytic therapy in the elderly. Thrombolytic therapy, in combination with either glycoprotein IIb/IIIa inhibitors or low-molecular weight heparin, poses an increased hazard in the elderly. Observational studies of thrombolytic therapy in the elderly portray a far worse outcome than the randomized clinical trials and raise the possibility of increased mortality. To date, no randomized trial has compared thrombolytic and primary coronary intervention in the elderly. However, multiple observational studies indicate a low risk of intracerebral hemorrhage and improved survival when a strategy of primary coronary intervention is employed. Future trials and observational studies should elucidate the ideal reperfusion strategy in the elderly. PMID- 12888707 TI - Elderly patients with acute coronary syndromes: higher risk and greater benefit from antiplatelet therapy and/or interventional therapies. AB - Outcomes in elderly patients with acute coronary syndromes are worse than in younger patients, and disappointingly, some therapies, such as thrombolysis for ST elevation myocardial infarction, appear to have less relative benefit than in younger patients. However, in unstable angina and non-ST elevation myocardial infarction, the elderly appear to derive greater relative and absolute benefit from the newer more potent antithrombotic therapies. With the glycoprotein IIb/IIIa inhibitors, an equivalent relative benefit has been observed, which translated into a greater absolute benefit in older vs. younger patients. Similarly, when comparing clopidogrel plus aspirin to aspirin alone, there was a consistent 20% reduction in cardiovascular death, myocardial infarction, or stroke in both elderly and younger patients. An emerging area of focus, however, is that of the appropriate dose in the elderly. Because the elderly on average have worse renal function, many drugs will not be cleared as well, and thus higher plasma levels will exist, which can translate into higher bleeding complications. Future studies are evaluating downward dose-adjustment of new therapies in the elderly as a means of improving the efficacy/safety profile. Thus, in unstable angina and non-ST elevation myocardial infarction, elderly patients are at higher risk and appear to derive particular benefit from more aggressive antithrombotic and interventional therapies. PMID- 12888708 TI - Ethical issues in the management of geriatric cardiac patients. AB - A 92-year-old patient dies less than a year after aortic valve replacement, pacemaker-AICD implantation, renal dialysis, and rehabilitation at a cost of over a half million dollars. PMID- 12888709 TI - Rate control compared to rhythm control in atrial fibrillation. PMID- 12888710 TI - Acute pericarditis: classic electrocardiogram. PMID- 12888711 TI - ALLHAT and ANBP2: what have we learned from recent mega-trials? PMID- 12888714 TI - Ocular surgical pharmacology: corneal endothelial safety and toxicity. AB - The purpose of this review is to discuss the recently published literature related to corneal endothelial toxicity and safety. We discuss postoperative complications, such as toxic endothelial cell destruction syndrome and toxic anterior segment syndrome, that cause significant injury to the patient and anxiety to the physician. Additionally, we review recent papers related to intraocular medications, preservatives, and devices, including antibiotics, anesthetics, viscoelastics, and enzymatic sterilization detergents, that have potentially toxic effects on the corneal endothelium. PMID- 12888715 TI - Surgical do's and don'ts of corneal dystrophies. AB - PURPOSE OF REVIEW: Characteristics of corneal dystrophies have been described with regards to such as location in the cornea, morphology, material composition, and recurrence after penetrating keratoplasty. The main goal of this review is to describe the surgical methods in treating corneal dystrophies. RECENT FINDINGS: Laser in situ keratomileusis (LASIK) has been shown to aggravate corneal deposits in Avellino dystrophy exacerbation LASIK and hence should be avoided. Phototherapeutic keratectomy (PTK) has shown its usefulness in clearing opacities with visual improvement and prevents painful erosion, resulting in delay or postponement of corneal grafting in some corneal dystrophies. Mitomycin-C may be used topically in conjunction with PTK to reduce the recurrence of the opacities. Topical use of antibody to TGF-beta can also be considered to suppress recurrence of corneal opacities after PTK or lamellar keratectomy. SUMMARY: Clinicians must become more adept at choosing a treatment depending on different genotypes and future studies on treatment of corneal dystrophies should be focused on establishing treatment of categorized corneal dystrophies based on their chromosomal mutation. PMID- 12888716 TI - Bioengineered corneas: how close are we? AB - Bioengineered corneas are substitutes for human donor tissue that are designed to replace part or the full thickness of damaged or diseased corneas. They range from prosthetic devices that solely address replacement of the cornea's function to tissue-engineered hydrogels that allow some regeneration of the host tissue. In addition, there are also bioengineered lenticules that may be implanted into the cornea to improve vision by altering the refractive properties of the eye, an alternative procedure to refractive surgery. In recent years, there have been significant developments in many areas of bioengineered corneas, such as the clinical trials of an artificial cornea designed as a prosthesis, the development of completely natural corneal replacements, and the development of biosynthetic matrices that permit host tissue regeneration. For correction of refractive errors, a synthetic corneal onlay that allows stable overgrowth of epithelium appears to be promising. PMID- 12888717 TI - Update on customized excimer ablations: recent developments reported in 2002. AB - Customized corneal ablation is an exciting frontier in refractive surgery that incorporates wavefront technology to detect and correct higher order aberrations in addition to spherocylindrical refractive errors. The goal is to achieve super normal vision in terms of acuity and contrast. As the concept of wavefront customized ablations is still new, there are a number of aspects of its clinical application that need analysis and understanding. Numerous reports have appeared in the literature during the past year that address the developments, concerns, and limitations of wavefront technology and custom ablation. We have attempted to summarize and discuss the significant reports in this current review. Our focus is on the optical and physiologic limits of wavefront customized correction, including the effect of accommodation, aging, and flap creation on the aberration profile. In addition, we also present the laser technology requirements, and clinical outcomes of customized excimer ablations that are reported to date. PMID- 12888718 TI - Update on nonexcimer laser refractive surgery technique: conductive keratoplasty. AB - Excimer laser-based refractive surgery techniques have been the most popular forms to correct myopia, hyperopia, and astigmatism. However, development of non excimer based refractive surgery such as thermal techniques provides a viable alternative to laser vision correction. Earlier forms of thermal techniques showed a lack of predictability and stability, resulting in the abandonment of the further development of these techniques. Recently, conductive keratoplasty, a laserless, radiofrequency-based technique, has been approved by the FDA for the correction of low to moderate hyperopia. Preliminary data showed that conductive keratoplasty seems to be safe, effective, and showed good refractive stability. This review will discuss recent studies on conductive keratoplasty technique in terms of its efficacy, safety, refractive predictability, and stability. PMID- 12888719 TI - Advances in subepithelial excimer refractive surgery techniques: Epi-LASIK. AB - The reports of an increasing incidence of iatrogenic ectasia, the evolution of wavefront aberrometry, and the suggestion that the laser in situ keratomileusis flap could lead to unpredictable biomechanical corneal changes have renewed interest in surface ablation and have set the stage for the introduction of alternative photorefractive treatment modalities. The theoretical advantage of surface procedures, such as laser epithelial keratomileusis that preserve the epithelial button, stems from the repositioning of the epithelial flap over the laser-ablated corneal surface. This epithelial sheet is thought to act as a natural contact lens that decreases postoperative pain and haze formation. Epipolis laser in situ keratomileusis is an alternative modality of epithelial separation with the use of a device that features a blunt oscillating blade. This surgical approach does not require the use of alcohol for epithelial loosening. PMID- 12888720 TI - Comparison of designs of laser systems utilized for refractive surgery. AB - Several different laser types are available on the market for the practitioner to use in refractive surgery. Each laser type has certain parameters that the surgeon must understand to obtain the best refractive outcomes. Studies published in peer-reviewed literature between February 2002 and March 2003 show that refractive results across all types of lasers have improved compared with those of several years ago. A difference in refractive outcomes between lasers is impossible to directly compare, as there are no randomized studies that hold surgeon or patient characteristics constant. PMID- 12888721 TI - Can you believe what you read? PMID- 12888724 TI - What's in a name? The taxonomy and nomenclature puzzle, part 2. AB - Learn the elements that make standardizing nursing nomenclatures and taxonomies a difficult task. PMID- 12888723 TI - Propelling your team to the top. AB - Learn the 10 benchmarks of forward movement for nurse managers. PMID- 12888725 TI - Retention reflects leadership style. PMID- 12888726 TI - Embracing National Patient Safety Goals, part 1 of 2. AB - Take an in-depth look at the National Patient Safety Goals to ensure your organization's compliance. PMID- 12888727 TI - Continuous assessment and regular communication foster patient safety. AB - Develop strategies to promote patient safety despite staffing crises. PMID- 12888728 TI - Infection control extra: combat West Nile virus. AB - Find out how to identify West Nile virus and learn prevention strategies. PMID- 12888729 TI - Taking charge of ventilator-associated pneumonia. AB - Target average length of stay and treatment costs to develop best practices for care of ventilator-associated pneumonia patients. PMID- 12888730 TI - Clocking care hours with workload measurement tools. AB - This study compares the Resource Utilization Group Classification System mandated by the Centers for Medicare and Medicaid for use in long-term care facilities to other existing workload measurement models. PMID- 12888731 TI - Motivating LPNs. AB - Follow one facility's approach to developing an LPN clinical ladder program. PMID- 12888732 TI - Telemetry beyond the ICU. AB - A Texas hospital uses telemetric oxygenation monitoring on medical/surgical floors to decrease critical care admissions for patients at risk of respiratory compromise. PMID- 12888733 TI - Hands-on help: get connected with the right PC. AB - Find up-to-date information on technology that can help you and your staff become more efficient. PMID- 12888734 TI - Consider alternative technologies to maintain vascular access devices. AB - Investigate alternatives to heparin, such as valved infusion catheters and luer activated positive fluid displacement valves. PMID- 12888737 TI - Streamline change-of-shift report. AB - Evaluate and streamline change-of-shift report to enhance critical thinking. PMID- 12888738 TI - Over-the-counter omeprazole (prilosec OTC). PMID- 12888739 TI - Aprepitant (emend) for prevention of nausea and vomiting due to cancer chemotherapy. PMID- 12888740 TI - Hypoglycemia and hyperglycemia with fluroroquinolones. PMID- 12888741 TI - Not such an innocent cough. PMID- 12888742 TI - A call to respond, not to react. PMID- 12888743 TI - The Strategic National Stockpile: roles and responsibilities of health care professionals for receiving the stockpile assets. AB - In 1999, the US Congress created the National Pharmaceutical Stockpile Program (NPS) under the management of the Centers for Disease Control and Prevention (CDC) and the Department of Health and Human Services. On March 1, 2003, the National Pharmaceutical Stockpile Program became known as the Strategic National Stockpile (SNS) Program under the new Department of Homeland Security. The purpose of the SNS Program is to maintain a stockpile of pharmaceutical agents, vaccines, medical supplies, and equipment to augment state and local resources during a large-scale disaster or bioterrorism event. Upon request, the SNS Program will deliver materials anywhere in the United States within 12 or fewer hours. A brief overview of the SNS Program and the role of local and state health care providers for receiving the SNS assets are reviewed. PMID- 12888744 TI - Case report: caring for suspected severe acute respiratory syndrome (SARS) patients. PMID- 12888745 TI - Providing nursing care with federal disaster-relief teams. PMID- 12888746 TI - Disaster preparedness in Virginia Hospital Center-Arlington after Sept 11, 2001. AB - True disaster readiness and preparedness are not really measured until put to the test of a real disaster. The attacks of Sept 11, 2001, put 1 community hospital and its disaster plan to the ultimate test. The process the hospital used to review, evaluate, and change its former disaster plan and then implement a new plan on the basis of the response to Sept 11 terrorists attacks is described. Specific challenges faced by this community hospital, the importance of participation in a regional response effort, and how the hospital has readied itself in a new threat environment are addressed. Disasters affect hospitals of all sizes and geographic locations. Health care facilities need a strong framework with which to function independently, if only for a short time, until additional resources can be implemented. PMID- 12888747 TI - A primer on epidemiology with SARS as an example. PMID- 12888748 TI - Respiratory protection for severe acute respiratory syndrome (SARS). PMID- 12888760 TI - Assessment of a selective surveillance method for detecting nosocomial infections in patients in the intensive care department. AB - BACKGROUND: The implementation of a time- and cost-effective system for the surveillance of the nosocomial infection (NI) is a challenge for infection control practitioners. OBJECTIVES: The aim of this study was to assess the sensitivity and the time reduction using a selective surveillance method (SSM) for the detection of NIs in comparison with a reference surveillance method (RSM). METHODS: During a 12-month period, surveillance was performed prospectively in 4 intensive care departments on a rotating basis. Using the RSM, NIs were identified by prospective chart reviews performed twice a week combined with weekly infectious disease ward rounds. In the SSM, surveillance was reduced to microbiologic data and participation in the weekly infectious disease ward rounds followed by selective chart review. RESULTS: In all, 578 patients amounting to 3597 patient-days were included in the study. In total, 78 NIs among 56 patients were identified. The overall sensitivity of the SSM compared with the RSM was 93.6% (73 of 78 NIs). The sensitivity of the SSM for the most important device-associated NIs (pneumonia, bloodstream infections, and urinary tract infections) was 96.3% (52 of 54 NIs) and 87.5% (21 of 24 NIs) for other NIs. Time required using the SSM was 1.3 hours compared with 4.1 hours per 10 beds per week (P =.0001) with the RSM. CONCLUSIONS: Within our setting, a SSM with restriction to microbiology reports and participation in the infectious disease ward rounds detected NIs with a high sensitivity and a remarkable time reduction. PMID- 12888761 TI - The state of infection surveillance and control in Canadian acute care hospitals. AB - BACKGROUND: Nosocomial infections and antibiotic-resistant pathogens cause significant morbidity, mortality, and economic costs. The infection surveillance and control resources and activities in Canadian acute care hospitals had not been assessed in 20 years. METHODS: In 2000, surveys were mailed to infection control programs in all Canadian hospitals with more than 80 acute care beds. The survey was modeled after the US Study on the Efficacy of Nosocomial Infection Control instrument, with new items dealing with resistant pathogens and computerization. Surveillance and control indices were calculated. RESULTS: One hundred seventy-two of 238 (72.3%) hospitals responded. In 42.1% of hospitals, there was fewer than 1 infection control practitioner per 250 beds. Just 60% of infection control programs had physicians or doctoral professionals with infection control training who provided services. The median surveillance index was 65.6/100, and the median control index was 60.5/100. Surgical site infection rates were reported to individual surgeons in only 36.8% of hospitals. CONCLUSIONS: There were deficits in the identified components of effective infection control programs. Greater investment in resources is needed to meet recommended standards and thereby reduce morbidity, mortality, and expense associated with nosocomial infections and antibiotic-resistant pathogens. PMID- 12888762 TI - Surgical site infections in patients undergoing major operations in a university hospital: using standardized infection ratio as a benchmarking tool. AB - BACKGROUND: Because patterns of infection acquired in patients undergoing operation are ever changing, it is an essential part of nosocomial infection surveillance programs to periodically document the epidemiologic features of infection in these patients. This study was conducted with the primary intention of describing the incidence and risk factors of the surgical site infection (SSI). METHODS: We performed a prospective study in patients undergoing certain major operations at a 750-bed university hospital in Thailand. The National Nosocomial Infection Surveillance (NNIS) system method and criteria were used for identifying and diagnosing infection. The infection rates were benchmarked with the NNIS report by means of indirect standardization and reported in terms of standardized infection ratio. Risk factors for SSI were evaluated using the multiple logistic regression model. RESULTS: From September 1998 to March 2000, the study included 4193 patients with 4437 major operations. The study identified 192 SSIs, 76 urinary catheter-related urinary tract infections, 26 central line related bloodstream infections, and 39 instances of ventilator-associated pneumonia (VAP), yielding an infection rate of 4.3 SSIs/100 operations, 11.0 catheter-related urinary tract infections/1000 urinary catheter-days, 6.1 central line-related bloodstream infections/1000 central line-days, and 11.0 VAPs/1000 ventilator-days. When compared with data from NNIS, the standardized infection ratio of SSI, catheter-related urinary tract infection, central line-related bloodstream infection, and VAP were 2.3, 2.1, 1.1, and 0.8, respectively. The factors that significantly associated with SSI were duration of operation in minutes, American Society of Anesthesiologists (ASA) class, and degree of wound contamination. CONCLUSION: All of the infection rates identified, except VAP, were higher than the average NNIS rates. The risk factors for SSI were prolonged duration of operation, poor physical status according to ASA classification, and higher degree of wound contamination. PMID- 12888763 TI - Spectrum of microbes and antimicrobial resistance in a surgical intensive care unit, Barbados. AB - BACKGROUND: To survey the epidemiologic findings of infections and antibiotic resistance patterns in the surgical intensive care unit (ICU) of a tertiary care university teaching hospital. METHODS: The microbiologic culture-sensitivity reports of patients admitted to a surgical ICU were prospectively studied for 6 months each of 3 consecutive years. The antibiotic usage for these patients also was studied concurrently. Reports from general surgical wards for 6 months of 1 year also were analyzed for comparison. The common specimens assayed microbiologically were tracheal aspirate, urine, blood, wound swabs, invasive catheter tips, and screening swabs for methicillin-resistant Staphylococcus aureus. RESULTS: The organisms reported were Enterobacteriaceae, Pseudomonas species, S aureus, and enterococci. Organisms were highly resistant to amoxicillin and first-generation cephalosporins because of the wide use of these drugs in the hospital. Pseudomonas species showed a 25% increase in resistance to piperacillin-tazobactam and an 18% increase to ciprofloxacin, which was correlated with the increased use of these antimicrobial agents (82% and 200% increases, respectively) in the unit during the 3 years. There was no increase in the resistance to ceftazidime because it is used less often. The resistance to ciprofloxacin, piperacillin-tazobactam, and ceftazidime was significantly greater in the ICU than in the general surgical wards in the same study period. CONCLUSIONS: The study provided data of antimicrobial resistance in a developing country with tourism as the main industry for epidemiologic comparison with other countries. PMID- 12888764 TI - Effect of intensive surveillance on cesarean-section wound infection rate in a Saudi Arabian hospital. AB - Surveillance for surgical site infection (SSI) in cesarean-section (C-section) in our hospital, carried out between September 1998 and July 1999, identified areas of deficiency in aseptic techniques that have contributed to increased incisional surgical site infection rates (ISSI) rates. On the basis of these findings, we intensified the infection control presence in this area and increased the number of in-services to our staff, with attention to infection control practices. Our senior infection control staff carried out ongoing monitoring of the ISSI rates as well as careful observation of aseptic technique. The aim was to reduce the rates of C-section ISSI in our hospital. We used the Centers for Disease Control and Prevention definitions for infection and the National Nosocomial Infections Surveillance System (NNISS) risk index in measuring the ISSI rates in the 2 time periods. We compared the ISSI rates with those of our preintervention period in an attempt to evaluate our interventional measures. Eight hundred seventy-five (875) C-sections were performed at King Fahad National Guard Hospital between January and December 2000. The overall ISSI rate was 1.37%. The ISSI rate for the NNISS risk category 0 (zero) was 1.2% and for the NNISS risk category 1 was 4.1%. There was a 50% reduction in the overall ISSI rates from our preintervention period (P <.05). Even though there were no objective means by which we measured the effect of infection control presence on the asepsis practices, there were no other changes that could have attributed to this significant reduction in ISSI rates in our C-section population. PMID- 12888765 TI - Nosocomial infections in medical-surgical intensive care units in Argentina: attributable mortality and length of stay. AB - BACKGROUND: Nosocomial infections are an important public health problem in many developing countries, particularly in the intensive care unit (ICU). Limited data exists on the incidence and burden of nosocomial infection in the ICU in Argentina. METHODS: We performed baseline prospective nosocomial infection surveillance of all patients for 6 months in 3 medical-surgical ICUs (MS-ICUs) in Argentina (2 months in each ICU). Nosocomial infections were identified using the Centers for Disease Control and Prevention National Nosocomial Infections Surveillance definitions. Overall and site-specific nosocomial infection rates, attributable mortality, and excess length of hospital stay were calculated. RESULTS: The overall nosocomial infection rate was 27% and 90 per 1000 patient days. The most common site of infection was catheter-related bloodstream infection (32%), followed by ventilator-associated pneumonia (25%), and catheter associated urinary tract infection (23%). The rate of central catheter-associated bloodstream infection in the MS-ICU was 44.61 per 1000 device-days, with an attributable mortality of 25%, and 12 attributable extra days of hospital stay. The urinary catheter-associated urinary tract infection rate in the MS-ICU was 22.55 per 1000 urinary catheter-days, with an attributable mortality of 5%, and 5 excess extra days of hospital stay. The ventilator-associated pneumonia rate in the MS-ICU was 50.87 per 1000 ventilator-days with an attributable mortality of 35%, and 10 attributable extra days of hospitalization. CONCLUSION: Our study finds high rates of nosocomial infections in ICUs in Argentina, associated with a considerable attributable mortality and excess length of stay. Ongoing targeted surveillance and implementation of infection control strategies is necessary to control this growing problem. PMID- 12888766 TI - Surgical wound infection surveillance in general surgery procedures at a teaching hospital in Pakistan. AB - BACKGROUND: A surveillance system was established at the Aga Khan University Hospital in Karachi, Pakistan, to determine surgical wound infection (SWI) rates, trends, and risk factors; and to compare rates with those reported by the National Nosocomial Infection Surveillance (NNIS) system of the Centers for Disease Control and Prevention. METHODS: Surveillance was performed from January 1997 to December 1999. Risk categorization was on the basis of the NNIS system. P <.05 was set for statistically significant difference between groups. Data were analyzed using the Epi-Info software (version 6.04, CDC, Atlanta, Ga). RESULTS: Overall SWI rates for the NNIS risk categories 0, 1, 2, and 3 were 1.9%, 3.7%, 6.7%, and 5.1%, respectively. SWI rate in 0 risk category decreased from 3% in 1997 to 1.1% in 1999 (P =.06). Multivariate analysis showed that SWI rates were higher after mastectomy (odds ratio [OR] 4.28, 95% confidence interval [CI] 1.8 10), hernia repair (OR 3.28, 95% CI 1.6-6.7), gastrointestinal resection (OR 2.2, 95% CI 0.88-5.9), skin procedures (OR 1.97, 95% CI 0.89-4.3), appendectomy OR 0.57, 95% CI 0.20-1.60, and miscellaneous procedures (OR 3.6, 95% CI 1.6-7.7), as compared with cholecystectomy. Other risk factors were contaminated type of operation (OR 2.6, 95% CI 1.2-5.5), and duration of operation exceeding the NNIS standard of "T" hours (OR 2.6, 95% CI 1.7-4). CONCLUSION: The SWI rates at the Aga Khan University Hospital are higher than the NNIS standards. There was a downward trend in the SWI rates during the surveillance period. A decrease in the duration of surgical procedures could further reduce the risk. PMID- 12888767 TI - A prospective study of surgical site infections in a pediatric hospital in Mexico City. AB - BACKGROUND: Pediatric surgical site infection (SSI) rates in the United States range from 2.5% to 4.4%. There is little data regarding their risk factors among children. We quantified SSI rates and identified risk factors of SSI in a tertiary care pediatric teaching hospital in Mexico City. METHODS: All neurosurgical, cardiovascular, and general surgical patients who underwent operation between Aug 1, 1998, and Jan 31, 1999, were followed-up daily during hospitalization. On postoperative day 30, a full review of microbiology reports and medical records was performed. Univariate and multivariate analyses were done to identify risk factors. RESULTS: Four hundred twenty-eight of 530 children completed follow-up. The overall SSI rate was 18.7%. Forty percent of SSI were superficial incisional, 21% were deep incisional, and 39% were organ/space infections. For clean, clean-contaminated, contaminated, and dirty procedures, SSI infection rates were 12.4%, 24.4%, 14.3%, and 32.4%, respectively. Open drains (OR = 2.3; 95% CI = 1.3-4.2; P <.005) and surgery that lasted 90 or more minutes (OR = 2.9; 95% CI = 1.6-5.1; P <.001) were associated with infection. CONCLUSIONS: Our rates are greater than comparable reported data among children. Duration of surgery and use of open drains were associated with SSI. PMID- 12888769 TI - Five years working with the German nosocomial infection surveillance system (Krankenhaus Infektions Surveillance System). AB - A national surveillance system for nosocomial infections, Krankenhaus Infektions Surveillance System (KISS), was established in Germany as a joint effort by the National Reference Center for Hospital Epidemiology (currently called the National Reference Center for the Surveillance of Nosocomial Infections) and the Robert Koch Institute. To begin with a well-accepted and proven method, surveillance protocols were developed on the basis of the National Nosocomial Infections Surveillance system. Hospitals from all parts of Germany took part voluntarily. There were 4 surveillance components: intensive care departments (227), patients undergoing operation (231 departments), neonatal intensive care departments (23 hospitals), and patients undergoing bone marrow and peripheral blood stem cell transplants (10 hospitals). Five years after the introduction of KISS many hospitals have introduced continuous surveillance activities whereby the various definitions and methods have attained wide acceptance. This has lead to much better understanding and cooperation between clinicians and infection control personnel in preventing nosocomial infections. KISS is considered the national reference database, but continuous evaluation and development of KISS are necessary to react adequately. PMID- 12888770 TI - Listeria monocytogenes cross-contamination in a nursery [corrected]. AB - Molecular evidence of Listeria monocytogenes cross-contamination in a nursery is presented. Listeria monocytogenes serotype 4b was isolated from the blood and the conjunctiva of a baby with neonatal sepsis who was born after septic amnionitis and premature rupture of membrane. Nine days later, the same bacterium was isolated from the cerebrospinal fluid of a second baby presenting with meningitis. Cervical cultures from the second baby's healthy mother were negative for Listeria sp. An in-depth epidemiologic investigation revealed that the same nurse administered routine treatments to both babies in the nursery during a 1 hour interval of time [corrected]. Pulse-field gel electrophoresis analysis of both strains with 2 different restriction enzymes demonstrated that they were identical and differ from other wild strains of L monocytogenes serotype 4b isolated in Israel. This fact strongly suggests that the second baby was infected during admittance to the nursery as a result of a hospital cross-contamination. PMID- 12888771 TI - Perceptions of physicians on infection control activities in Turkish hospitals: a cross-sectional survey. AB - BACKGROUND: Most countries practice some form of infection control (IC) to minimize hospital-acquired infections. There is still considerable diversity in IC activities in Turkey. Little is known about practical issues and perceptions of physicians regarding IC in Turkey. METHODS: The authors developed an 11-item questionnaire addressing questions on IC activities. Physicians attending a national infection control meeting (n = 285) were asked to participate. RESULTS: The response rate was 86%. Respondents from nonuniversity teaching hospitals stated a higher involvement in IC. Physicians from university hospitals were in greater need of IC training. CONCLUSIONS: Academic hospitals are particularly concerned with IC. Domains such as surveillance of endemic hospital infections, specific preventive protocols, antibiotics policy, and staff education should be addressed by IC committees. IC is well established in Turkey, but there is still a long way to go to reach the standards found in developed countries. PMID- 12888772 TI - Chronic pubertal, but not adult chronic cannabinoid treatment impairs sensorimotor gating, recognition memory, and the performance in a progressive ratio task in adult rats. AB - There is evidence from studies in humans and animals that a vulnerable period for chronic cannabinoid administration exists during certain phases of development. The present study tested the hypothesis that long-lasting interference of cannabinoids with the developing endogenous cannabinoid system during puberty causes persistent behavioral alterations in adult rats. Chronic treatment with the synthetic cannabinoid agonist WIN 55,212-2 (WIN) (1.2 mg/kg) or vehicle was extended over 25 days either throughout the rats' puberty or for a similar time period in adult rats. The rats received 20 injections intraperitoneally (i.p.), which were not delivered regularly. Adult rats were tested for object recognition memory, performance in a progressive ratio (PR) operant behavior task, locomotor activity, and prepulse inhibition (PPI) of the acoustic startle response (ASR). PPI was significantly disrupted only by chronic peripubertal cannabinoid treatment. This long-lasting PPI deficit was reversed by the acute administration of the dopamine antagonist haloperidol. Furthermore, we found deficits in recognition memory of pubertal-treated rats and these animals showed lower break points in a PR schedule, whereas food preference and locomotion were not affected. Adult chronic cannabinoid treatment had no effect on the behaviors tested. Therefore, we conclude that puberty in rats is a vulnerable period with respect to the adverse effects of cannabinoid treatment. Since PPI deficits, object recognition memory impairments, and anhedonia/avolition are among the endophenotypes of schizophrenia, we propose chronic cannabinoid administration during pubertal development as an animal model for some aspects of the etiology of schizophrenia. PMID- 12888773 TI - Methyl nicotinate-induced vasodilation in generalized social phobia. AB - Elevated vasodilatory response (blushing) to social situations is characteristic of social phobia (SP). A relatively unexplored basis for this phenomenon is alteration in underlying vasodilatory mechanisms. To investigate this possibility, we evaluated the vasodilatory response induced by methyl nicotinate (niacin ester derivative) in 31 generalized SP patients and 41 matched healthy volunteers (HV). A patch impregnated with 0, 0.1, 0.5, 1, and 10 mM methyl nicotinate was applied to the forearm or face of subjects for 1 min, followed by 20-min laser Doppler spectroscopy blood flow monitoring. Blood flow stimulation with 1 and 10 mM methyl nicotinate was significantly reduced in SP patients by 35 and 17%, respectively. Induced blood flow was negatively correlated with patients' Leibowitz Social Phobia Scale (LSAS) at 1 and 10 mM doses. Furthermore, the maximal rate of change of vasodilatory reaction was correlated with symptom scores at 1 and 10 mM doses. Induced increases in the arm and face blood flow measurements correlated, supporting the external validity of the former location. Generalized SP patients vasodilate less to topical methyl nicotinate challenges, with effect amplification in severely ill patients. Although the mechanism for this is unclear, we propose desensitization of the prostaglandin-mediated vasodilating system as an explanation. Neuropsychopharmacology (2003) 28, 1846 1851, advance online publication, 23 July 2003; doi:10.1038/sj.npp.1300227 PMID- 12888774 TI - Manipulations during the second, but not the first, week of life increase susceptibility to cocaine self-administration in female rats. AB - We compared the effects of manipulations during week 1 vs week 2 of life on the propensity to self-administer cocaine. Pups received daily subcutaneous saline injections, were handled briefly, or remained undisturbed during their respective treatment periods. Animals handled during the second week of life exhibited increased locomotor response to novelty when tested on postnatal day (PND) 48, compared to all other groups. Rats were implanted with jugular catheters on PND 70 and then given the opportunity to self-administer (0.125 mg/kg/infusion) cocaine for 5 consecutive days (1 h sessions). The dose was then raised to 0.25 mg/kg/infusion for 5 days and to 0.5 mg/kg/infusion for the final 5 days of testing. Only animals manipulated during the second week of life acquired drug taking behavior. These effects were both stimulus- and gender-specific. Females handled during the second week of life acquired cocaine self-administration (SA) at the lowest dose, and females injected during the second week of life acquired at the intermediate dose. Males injected during the second week of life showed a similar, but more variable, drug-taking pattern. There were no group differences in serum corticosterone response to novelty, although relative to undisturbed animals and those manipulated in the first week of life, female animals manipulated during the second week of life had lower basal expression of hippocampal glucocorticoid receptor mRNA in adulthood. We conclude that the second week of life in the rodent is a sensitive period during which manipulations result in a more vulnerable phenotype for the acquisition of cocaine SA. PMID- 12888776 TI - Variations in maternal care alter GABA(A) receptor subunit expression in brain regions associated with fear. AB - Maternal care influences the development of stress reactivity in the offspring. These effects are accompanied by changes in corticotropin-releasing factor (CRF) expression in brain regions that regulate responses to stress. However, such effects appear secondary to those involving systems that normally serve to inhibit CRF expression and release. Thus, maternal care over the first week of life alters GABA(A) (gamma-aminobutyric acid)(A) receptor mRNA subunit expression. The adult offspring of mothers that exhibit increased levels of pup licking/grooming and arched back-nursing (high LG-ABN mothers) show increased alpha1 mRNA levels in the medial prefrontal cortex, the hippocampus as well as the basolateral and central regions, of the amygdala and increased gamma2 mRNA in the amygdala. Western blot analyses confirm these effects at the level of protein. In contrast, the offspring of low LG-ABN mothers showed increased levels of alpha3 and alpha4 subunit mRNAs. The results of an adoption study showed that the biological offspring of low LG-ABN mothers fostered shortly after birth to high LG-ABN dams showed the increased levels of both alpha1 and gamma2 mRNA expression in the amygdala in comparison to peers fostered to other low LG-ABN mothers (the reverse was true for the biological offspring of high LG-ABN mothers). These findings are consistent with earlier reports of the effects of maternal care on GABA(A)/benzodiazepine receptor binding and suggest that maternal care can permanently alter the subunit composition of the GABA(A) receptor complex in brain regions that regulate responses to stress. PMID- 12888775 TI - Modulation of mood and cognitive performance following acute administration of single doses of Melissa officinalis (Lemon balm) with human CNS nicotinic and muscarinic receptor-binding properties. AB - Melissa officinalis (Lemon balm) is a herbal medicine that has traditionally been attributed with memory-enhancing properties, but which is currently more widely used as a mild sedative and sleep aid. In a previous study it was demonstrated that a commercial Melissa extract led to dose-specific increases in calmness, and dose-dependent decrements in timed memory task performance. However, the extract utilized in that study did not exhibit in vitro cholinergic receptor-binding properties. The current study involved an initial screening of samples of M. officinalis for human acetylcholinesterase inhibition and cholinergic receptor binding properties. The cognitive and mood effects of single doses of the most cholinergically active dried leaf were then assessed in a randomized, placebo controlled, double-blind, balanced crossover study. Following the in vitro analysis, 20 healthy, young participants received single doses of 600, 1000, and 1600 mg of encapsulated dried leaf, or a matching placebo, at 7-day intervals. Cognitive performance and mood were assessed predose and at 1, 3, and 6 h postdose using the Cognitive Drug Research computerized assessment battery and Bond-Lader visual analog scales, respectively. In vitro analysis of the chosen extract established IC(50) concentrations of 0.18 and 3.47 mg ml(-1), respectively, for the displacement of [(3)H]-(N)-nicotine and [(3)H]-(N) scopolamine from nicotinic and muscarinic receptors in the human cerebral cortex tissue. However, no cholinesterase inhibitory properties were detected. The most notable cognitive and mood effects were improved memory performance and increased 'calmness' at all postdose time points for the highest (1600 mg) dose. However, while the profile of results was overwhelmingly favorable for the highest dose, decrements in the speed of timed memory task performance and on a rapid visual information-processing task increased with decreasing dose. These results suggest that doses of Melissa officinalis at or above the maximum employed here can improve cognitive performance and mood and may therefore be a valuable adjunct in the treatment of Alzheimer's disease. The results also suggest that different preparations derived from the same plant species may exhibit different properties depending on the process used for the sample preparation. PMID- 12888777 TI - Harm avoidance, anxiety, and response to novelty in the adolescent S-100beta transgenic mouse: role of serotonin and relevance to Down syndrome. AB - S-100beta is an astroglial-derived protein, which plays a role in brain development and maintenance, and is known to play a specific role in the regulation of growth of the serotonergic neuronal system. In humans, the gene for S-100beta is found on chromosome 21, within the region that is considered important for the phenotype of Down syndrome (DS). Thus, we have been studying a model of DS, the S-100beta transgenic mouse. In the current study, we have examined anxiety and responses to novelty in adolescent (60-90 days) animals, at a time when we have shown the animals to be relatively lacking in serotonin innervation, compared to their CD-1 nontransgenic controls. In a test for approach/avoidance, the light/dark test, the S-100beta transgenic mice animals showed no differences from control CD-1 mice. However, in the hole-board test for exploratory behavior, the S-100beta animals were found to be less responsive to the inhibiting effects of the serotonin receptor 5-HT1A agonist, buspirone. Three tests were used to measure response to novelty. In the open field, the S-100beta animals showed greater activity longer than the control animals, and in the Y maze test, the S-100beta animals spent more time in the novel arm. In a test for novelty-induced gnawing, the S-100beta animals were also more active than control animals. All of these suggest that the S-100beta transgenic mice are slower to habituate to novelty than control animals. Finally, we tested the animals in a new procedure that we are proposing as a test for harm avoidance. In this apparatus, the S-100beta animals showed more approaches to a novel and potentially harmful object than the control mice did. These results are discussed in reference to the known lack of serotonin in the animals, and to the behavioral phenotype of DS. PMID- 12888778 TI - Altered NMDA glutamate receptor antagonist response in recovering ethanol dependent patients. AB - Ethanol is an antagonist of the N-methyl-D-aspartate (NMDA) glutamate receptor. Ethanol dependence upregulates NMDA receptors and contributes to crosstolerance with selective NMDA receptor antagonists in animals. This study evaluated whether recovering ethanol-dependent patients show evidence of a reduced level of response to the effects of the NMDA receptor antagonist, ketamine. In this double blind study, 34 recently detoxified alcohol-dependent patients and 26 healthy comparison subjects completed 3 test days involving a 40-min infusion of saline, ketamine 0.1 mg/kg, or ketamine 0.5 mg/kg in a randomized order. Recovering ethanol-dependent patients showed reduced perceptual alterations, dysphoric mood, and impairments in executive cognitive functions during ketamine infusion relative to the healthy comparison group. No attenuation of ketamine-induced amnestic effects, euphoria, or activation was observed. The alterations in NMDA receptor function observed in recovering ethanol-dependent patients may have important implications for ethanol tolerance, ethanol dependence, and the treatment of alcoholism. PMID- 12888779 TI - ABCB1 (MDR1)-type P-glycoproteins at the blood-brain barrier modulate the activity of the hypothalamic-pituitary-adrenocortical system: implications for affective disorder. AB - Multidrug-resistance gene 1-type P-glycoproteins (ABCB1-type P-gps) protect the brain against the accumulation of many toxic xenobiotics and drugs. We recently could show that the access of the endogenous glucocorticoids corticosterone and cortisol to the brain are regulated by ABCB1-type P-gps in vivo. ABCB1-type P-gp function, therefore, is likely to exert a profound influence on the regulation of the hypothalamic-pituitary-adrenocortical (HPA) system. Hyperactivity of the HPA system is frequently observed in human affective disorder, and a considerable amount of evidence has been accumulated suggesting that normalization of the HPA system might be the final step necessary for stable remission of the disease. To examine whether blood-brain barrier (BBB) function influences neuroendocrine regulation, we investigated HPA system activity in abcb1ab (-/-) mice under basal conditions and following stress. Abcb1ab (-/-) mice showed consistently lower plasma ACTH levels and lower evening plasma corticosterone levels. CRH mRNA expression in the hypothalamic paraventricular nucleus was decreased and pituitary POMC mRNA expressing cells were significantly reduced in number in abcb1ab (-/-) mutants; however, they showed a normal activation of the HPA system following CRH stimulation. Lower doses of dexamethasone were required to suppress plasma corticosterone levels in mutants. Our data thus provide evidence for a sustained suppression of the HPA system at the hypothalamic level in abcb1ab (-/ ) mice, suggesting that BBB function significantly regulates HPA system activity. Whether naturally occurring polymorphisms in the human ABCB1 gene might result in persistent changes in the responsiveness and regulation of the HPA system will be the subject of future investigations, correlating both genetic information with individual characteristics of the neuroendocrine phenotype. PMID- 12888780 TI - Positron emission tomography studies of potential mechanisms underlying phencyclidine-induced alterations in striatal dopamine. AB - Positron emission tomography (PET), in combination with (11)C-raclopride, was used to examine the effects of phencyclidine (PCP) on dopamine (DA) in the primate striatum. In addition, we explored the hypotheses that GABAergic pathways as well as molecular targets beyond the N-methyl-D-aspartate (NMDA) receptor complex (ie dopamine transporter proteins, DAT) contribute to PCP's effects. In the first series of experiments, (11)C-raclopride was administered at baseline and 30 min following intravenous PCP administration. In the second series of studies, gamma-vinyl GABA (GVG) was used to assess whether enhanced GABAergic tone altered NMDA antagonist-induced changes in DA. Animals received an initial PET scan followed by pretreatment with GVG (300 mg/kg), then PCP 30 min prior to a second scan. Finally, we explored the possible contributions of DAT blockade to PCP-induced increases in DA. By examining (11)C-cocaine binding a paradigm in which PCP was coadministered with the radiotracer, we assessed the direct competition between these two compounds for the DAT. At 0.1, 0.5, and 1.0 mg/kg, PCP decreased (11)C-raclopride binding by 2.1, 14.9+/-2.2 and 8.18+/-1.1%, respectively. These effects were completely attenuated by GVG (3.38+/-3.1% decrease in (11)C-raclopride binding). Finally, PCP (0.5 mg/kg) decreased (11)C cocaine binding by 25.5+/-4.3%, while at 1.0 mg/kg this decrease was 13.5%, consistent with a competitive interaction at the DAT. These results suggest that PCP may be exerting some direct effects through the DAT and that GABA partially modulates NMDA-antagonist-induced increases in striatal DA. PMID- 12888781 TI - Olanzapine reduces craving for alcohol: a DRD4 VNTR polymorphism by pharmacotherapy interaction. AB - Separate investigations have suggested that olanzapine, a D4 antagonist, decreases craving after a priming dose of alcohol and that the DRD4 variable number of tandem repeats (VNTR) polymorphism influences the expression of craving after a priming dose of alcohol. The present study tested the hypothesis that olanzapine may be differentially effective at reducing cue-elicited craving based on individual differences in DRD4 VNTR in a sample of heavy social drinkers. Participants were randomly assigned to receive olanzapine (5 mg) or a control medication (cyproheptadine, 4 mg) prior to consuming three alcoholic drinks. Participants completed subjective measures of craving and euphoria after each drink. Participants who were homozygous or heterozygous for the 7 (or longer) repeat allele of the DRD4 VNTR were classified as DRD4 L, while the other participants were classified as DRD4 S. The findings indicated that olanzapine reduces craving for alcohol at baseline for both DRD4 S and DRD4 L individuals, but only reduces craving after exposure to alcohol cues and after a priming dose of alcohol for DRD4 L individuals. PMID- 12888782 TI - L-tryptophan challenge and cognitive deficits in bipolar disorder: evidence for hyperserotonergic and hypodopaminergic mechanisms. PMID- 12888783 TI - Subdissociative dose ketamine produces a deficit in manipulation but not maintenance of the contents of working memory. AB - We investigated the effects of subdissociative dose ketamine on executive processes during a working memory task. A total of 11 healthy volunteers participated in a double-blind, placebo-controlled, randomized, within-subjects study. They attended on three occasions, receiving intravenous infusions of placebo, a lower ketamine dose, and a higher ketamine dose. On each occasion, they underwent a series of tasks engaging working memory function in verbal and visuo-spatial domains. Further tasks explored aspects of long-term memory, planning, attention, and perceptual processing. With respect to working memory/executive function, a highly specific pattern of impairment was observed. Impairments were seen only at the higher dose of ketamine and restricted to a subgroup of the verbal working memory tasks: While visuo-spatial working memory showed no evidence of impairment, and while simple maintenance processes during verbal working memory were also unimpaired, higher dose ketamine produced a significant impairment in the manipulation of information within working memory. This process-specific effect of ketamine was reflected in a drug-by-task interaction. The specificity of this ketamine effect suggests that the earliest effect of NMDA receptor blockade is in higher order control of executive function rather than in more basic maintenance processes. PMID- 12888784 TI - Differential role of serotonergic projections arising from the dorsal and median raphe nuclei in locomotor hyperactivity and prepulse inhibition. AB - While an involvement of brain serotonin systems in schizophrenia has been suggested by many studies, the relative role of different serotonergic projections in the brain remains unclear. We therefore examined the effects of selective brain serotonin depletion on psychotropic drug-induced locomotor hyperactivity and prepulse inhibition, two animal models of aspects of schizophrenia. Pentobarbital-anesthetized (60 mg/kg, i.p.) male Sprague-Dawley rats were stereotaxically microinjected with 1 microl of a 5 microg/microl solution of the serotonergic neurotoxin 5,7-dihydroxytryptamine (5,7-DHT) into either the dorsal or median raphe nucleus. At 2 weeks after the surgery, rats with dorsal raphe lesions did not show changes in psychotropic drug-induced locomotor hyperactivity, but displayed partial disruption of prepulse inhibition. In contrast, rats with median raphe lesions showed significant enhancement of phencyclidine-induced, but not amphetamine-induced locomotor hyperactivity and a marked disruption of prepulse inhibition. These results provide evidence for differential involvement of serotonergic projections in locomotor hyperactivity and prepulse inhibition. This study may help to explain the role of different serotonin projections in the brain in the pathophysiology of schizophrenia. PMID- 12888786 TI - Efficient T cell receptor-mediated apoptosis in nonobese diabetic mouse thymocytes. PMID- 12888788 TI - Stopping HIV before it begins: issues faced by women in India. AB - Despite advances in understanding HIV pathogenesis, the economic, social and legal constraints in India continue to make women particularly vulnerable to HIV infection. PMID- 12888789 TI - Toll gets tied in a knot. PMID- 12888790 TI - Rap1 redux. PMID- 12888791 TI - Deadbeat neighbors influence thymocyte lineage commitment. PMID- 12888792 TI - Nefarious abuse. PMID- 12888794 TI - Antigen presentation to naive CD4 T cells in the lymph node. AB - Although the presentation of peptide-major histocompatibility complex class II (pMHC class II) complexes to CD4 T cells has been studied extensively in vitro, knowledge of this process in vivo is limited. Unlike the in vitro situation, antigen presentation in vivo takes place within a complex microenvironment in which the movements of antigens, antigen-presenting cells (APCs) and T cells are governed by anatomic constraints. Here we review developments in the areas of lymph node architecture, APC subsets and T cell activation that have shed light on how antigen presentation occurs in the lymph nodes. PMID- 12888795 TI - Data from the Northern Finland 1966 Birth Cohort support an association between depression and immune function. PMID- 12888798 TI - Lack of association of HOXA1 and HOXB1 mutations and autism in Sicilian (Italian) patients. PMID- 12888799 TI - Family-based association study of DTNBP1 in 6p22.3 and schizophrenia. PMID- 12888800 TI - Analysis of a polymorphism in the promoter region of the tumor necrosis factor alpha gene in schizophrenia and bipolar disorder: further support for an association with schizophrenia. PMID- 12888801 TI - GABAergic dysfunction in mood disorders. AB - The authors review the available literature on the preclinical and clinical studies involving GABAergic neurotransmission in mood disorders. Gamma aminobutyric acid (GABA) is an inhibitory neurotransmitter present almost exclusively in the central nervous system (CNS), distributed across almost all brain regions, and expressed in interneurons modulating local circuits. The role of GABAergic dysfunction in mood disorders was first proposed 20 years ago. Preclinical studies have suggested that GABA levels may be decreased in animal models of depression, and clinical studies reported low plasma and CSF GABA levels in mood disorder patients. Also, antidepressants, mood stabilizers, electroconvulsive therapy, and GABA agonists have been shown to reverse the depression-like behavior in animal models and to be effective in unipolar and bipolar patients by increasing brain GABAergic activity. The hypothesis of reduced GABAergic activity in mood disorders may complement the monoaminergic and serotonergic theories, proposing that the balance between multiple neurotransmitter systems may be altered in these disorders. However, low GABAergic cortical function may probably be a feature of a subset of mood disorder patients, representing a genetic susceptibility. In this paper, we discuss the status of GABAergic hypothesis of mood disorders and suggest possible directions for future preclinical and clinical research in this area. PMID- 12888802 TI - Atopy and depression: results from the Northern Finland 1966 Birth Cohort Study. AB - Several studies have suggested an association between IgE-mediated atopic allergies and depression. The present study extends our understanding about putative gender differences of this association and provides further epidemiological evidence for our previous finding that the association between atopy and depression may be characteristic for females only. In order to clearly determine the presence of atopic disorders and depression, we used more valid tools than had been employed earlier and we had access to a database (the Northern Finland 1966 Birth Cohort), in which individuals were followed up prospectively until the age of 31 years. The information on allergic symptoms, verified by skin-prick tests and comprising data of 5518 individuals, was used to ascertain the presence of atopy. Depression was assessed with the help of Hopkins' Symptom Checklist-25 and self-reported doctor-diagnosed depression. After adjusting for a father's social class, mother's parity, and place of residence, logistic regression analyses showed that the risk of developing depression increased in parallel with the increasing severity of depression and, when compared with nonatopic subjects, was 3.0 to 4.7-fold up in atopic females and statistically significant. In atopic males, the association between atopy and depression was statistically significant only in the highest depression scores, the odds ratio being 6.3-fold. The results indicate that females suffering from atopic diseases might possess an elevated risk of developing depression already during early adulthood. In males, the association between these two disorders is evident only among the most severe manifestations of depression. Possible background theories, that is, genetic abnormalities in serotonin metabolism, HPA axis dysfunction, and histamine theory are discussed. PMID- 12888803 TI - Met66 in the brain-derived neurotrophic factor (BDNF) precursor is associated with anorexia nervosa restrictive type. AB - Several lines of evidence support a role for brain-derived neurotrophic factor (BDNF) alterations in the etiology of eating disorders (EDs). BDNF heterozygous knockout mice show alterations in eating behavior, increased body weight and adipocyte hypertrophy. BDNF also regulates the synaptic efficiency through the modulation of key neurotransmitter systems previously known to be involved in ED. These findings, together with the fact that this neurotrophin is expressed in the hypothalamus nuclei associated with weight regulation and feeding control, led us to propose BDNF as a candidate gene for ED. To investigate the possible involvement of this neurotrophin in eating behavior, we screened the BDNF gene in 95 ED patients and identified four sequence variants. Two of them, -374A/T and 256G/A, were found in two patients with anorexia nervosa (AN) and consisted of single-nucleotide mutations within the 5' untranslated region (5'UTR). The other two polymorphisms resulted in a C to T transition located at the 5'UTR of the BDNF gene and an amino-acid substitution within the BDNF precursor protein (Val66Met). We performed a case-control study for these two Single-nucleotide polymorphisms in a sample of 143 ED patients and 112 unrelated controls and found a strong association of restricting AN (ANR) with the Met allele of the Val66Met BDNF polymorphism (2p=0.002). There was also evidence for a significant effect of this sequence variant on the minimum body mass index (MBMI) (2p=0.006). These results suggest that the BDNF Met66 variant may be a susceptibility factor to ED, mainly to ANR and low MBMI. PMID- 12888804 TI - Neuronal mechanism of nociceptin-induced modulation of learning and memory: involvement of N-methyl-D-aspartate receptors. AB - Nociceptin (also called orphanin FQ) is an endogenous heptadecapeptide that activates the opioid receptor-like 1 (ORL1) receptor. Nociceptin system not only affects the nociception and locomotor activity, but also regulates learning and memory in rodents. We have previously reported that long-term potentiation and memory of ORL1 receptor knockout mice are enhanced compared with those in wild type mice. Here, we show the neuronal mechanism of nociceptin-induced modulation of learning and memory. Retention of fear-conditioned contextual memory was significantly enhanced in the ORL1 receptor knockout mice without any changes in cued conditioned freezing. Inversely, in the wild-type mice retention of contextual, but not cued, conditioning freezing behavior was suppressed by exogenous nociceptin when it was administered into the cerebroventricle immediately after the training. ORL1 receptor knockout mice exhibited a hyperfunction of N-methyl-D-aspartate (NMDA) receptor, as evidenced by an increase in [3H]MK-801 binding, NMDA-evoked 45Ca2+ uptake and activation of Ca2+/calmodulin-dependent protein kinase II (CaMKII) activity and its phosphorylation as compared with those in wild-type mice. The NMDA-induced CaMKII activation in the hippocampal slices of wild-type mice was significantly inhibited by exogenous nociceptin via a pertussis toxin-sensitive pathway. However, the alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid receptor GluR1 subunit at Ser831 and Ser845, and NMDA receptor subunit NR2B at Thr286 were phosphorylated similarly after NMDA receptor stimulation in both type of mice. The expressions of GluR1 and GluR2 also did not change, but the levels of polysialylated form of neuronal cell adhesion molecule (N-CAM) were reduced in the ORL1 receptor knockout as compared with wild-type mice. These results suggest that nociceptin system negatively modulates learning and memory through the regulation of NMDA receptor function and the expression of N-CAM. PMID- 12888806 TI - Celebrating novel cancer drugs. PMID- 12888807 TI - Vascular endothelial growth factors C and D and lymphangiogenesis in gastrointestinal tract malignancy. AB - Vascular endothelial growth factor-C (VEGF-C) and VEGF-D are members of the VEGF family of cytokines and have angiogenic and lymphangiogenic actions. In gastric adenocarcinoma, VEGF-C mRNA and tissue protein expression correlate with lymphatic invasion, lymph node metastasis and in some reports, venous invasion and reduced 5-year survival. Patients with gastric adenocarcinomas containing high levels of VEGF-C expression have significantly reduced 5-year survival rates, and VEGF-C expression is an independent prognostic risk factor for death. The role of VEGF-C in oesophageal squamous and colorectal cancer and VEGF-D in colorectal cancer is not clear, with conflicting reports in the published literature. In order to exploit potential therapeutic applications, further research is necessary to define the precise roles of these cytokines in health and disease. PMID- 12888808 TI - Survival analysis part II: multivariate data analysis--an introduction to concepts and methods. PMID- 12888810 TI - Treatment of neuroendocrine tumours with infusional 5-fluorouracil, folinic acid and streptozocin. AB - A standardised, effective systemic therapy for metastatic neuroendocrine tumours (NETs) has not been established to date. We reviewed the management of 15 patients with inoperable, metastatic NET treated systematically with a combination chemotherapy regimen of infusional 5-fluorouracil, folinic acid and streptozocin. Overall objective response rate was 53% and tolerability was excellent. PMID- 12888811 TI - Does HIV adversely influence the outcome in advanced non-small-cell lung cancer in the era of HAART? AB - The objectives of the study are to assess the impact of HIV status on the outcome of patients with non-small-cell lung cancer (NSCLC) in the era of highly active antiretroviral therapy (HAART). Patients diagnosed with HIV-related NSCLC in the HAART era (since January 1996) were identified from a prospective single-centre lung cancer database. The clinicopathological characteristics and outcome of each HIV-positive patient were compared to three age- and stage-matched HIV-negative controls with NSCLC who were diagnosed over the same time period and treated in an identical manner. The results showed that the two groups had similar disease characteristics and received a similar amount of chemotherapy. The median overall survival of the two groups was the same (4 months, log rank P=0.55). None of the HIV-positive patients developed an AIDS defining illness or died of HIV during treatment or follow-up. In conclusion, in this cohort, HIV status does not influence the prognosis of advanced NSCLC. This suggests that the survival of patients with HIV-related NSCLC may have improved since the introduction of HAART, and this may be due to a decrease in HIV-related deaths. PMID- 12888809 TI - Professor Tom Connors and the development of novel cancer therapies by the Phase I/II Clinical Trials Committee of Cancer Research UK. PMID- 12888812 TI - Neoadjuvant, adjuvant and palliative treatment of gastrointestinal stromal tumours (GIST) with imatinib: a centre-based study of 17 patients. AB - Malignant gastrointestinal stromal tumours (GIST) have a poor prognosis. Since these tumours are resistant to conventional radiation and chemotherapy, surgery has been the mainstay of treatment. However, surgery is usually inadequate for the treatment of malignant GIST. Imatinib, a KIT tyrosine kinase inhibitor, has recently been found to have a dramatic antitumour effect on GIST. In this centre based study of 17 consecutive patients with high-risk or overtly malignant GIST, imatinib was used in three different settings - palliatively, adjuvantly, and neoadjuvantly. The treatment was found to be safe and particularly effective in tumours with activating mutations of exon 11 of the KIT gene. Clinical response to imatinib treatment correlated morphologically to tumour necrosis, hyalinisation, and reduced proliferative activity. The value of neoadjuvant imatinib treatment was illustrated in one case. PMID- 12888813 TI - A medical nutriment has supportive value in the treatment of colorectal cancer. AB - MSC (Avemar) is a medical nutriment of which preclinical and observational clinical studies suggested an antimetastatic activity with no toxicity. This open label cohort trial has compared anticancer treatments plus MSC (9 g once daily) vs anticancer treatments alone in colorectal patients, enrolled from three oncosurgical centres; cohort allocation was on the basis of patients' choice. Sixty-six colorectal cancer patients received MSC supplement for more than 6 months and 104 patients served as controls (anticancer therapies alone): no statistical difference was noted in the time from diagnosis to the last visit between the two groups. End-point analysis revealed that progression-related events were significantly less frequent in the MSC group (new recurrences: 3.0 vs 17.3%, P<0.01; new metastases: 7.6 vs 23.1%, P<0.01; deaths: 12.1 vs 31.7%, P<0.01). Survival analysis showed significant improvements in the MSC group regarding progression-free (P=0.0184) and overall survivals (P=0.0278) probabilities. Survival predictors in Cox's proportional hazards were UICC stage and MSC treatment. Continuous supplementation of anticancer therapies with MSC for more than 6 months is beneficial to patients with colorectal cancer in terms of overall and progression-free survival. PMID- 12888814 TI - Treatment of stage 4s neuroblastoma--report of 10 years' experience of the French Society of Paediatric Oncology (SFOP). AB - Stage 4s neuroblastoma (NB) is usually associated with a favourable outcome, despite a large tumour burden, as spontaneous regression frequently occurs. However, in some infants rapid disease progression can be observed with severe functional impairment. Thus, for all patients the potential risks of cytotoxic therapy must be weighed against the benefits of early medical intervention. We have retrospectively reviewed the charts of 94 infants treated for stage 4s NB in centres of the French Society of Paediatric Oncology between 1990 and 2000, and describe the different first-line treatment approaches that were, successively, liver irradiation, chemotherapy using a cyclophosphamide-vincristine regimen, and chemotherapy using a carboplatin-etoposide regimen. The overall survival was 88% (+/-7.6%), with a mean follow-up of 64 months. Elevated serum neuron-specific enolase (>100 nmol ml(-1)), ferritin (>280 ng ml(-1)) and urinary dopamine levels (>2500 nmol mmol(-1) creatinine) were associated with a poor outcome, as were the genetic markers N-myc amplification and chromosome 1p deletion (P<0.0005 and P=0.0016, respectively). Patients who required medical intervention at diagnosis fared worse than those who received supportive treatment only (P<0.005). The clinical evolution observed with the different successive treatment approaches suggests that if infants do require therapy, the prompt initiation of a more intensive regimen such as carboplatin-etoposide may be more beneficial. PMID- 12888815 TI - Hypersensitivity reactions related to oxaliplatin (OHP). AB - Patients treated with platinum compounds are subject to hypersensitivity reactions. Our study has highlighted the reactions related to oxaliplatin (OHP) infusion. One hundred and twenty-four patients affected by advanced colorectal cancer were treated with different schedules containing OHP, at the Institute of Haematology and Medical Oncology 'L. and A. Seragnoli' of Bologna and at the Medical Oncology Division of Livorno Hospital. Seventeen patients (13%) showed hypersensitivity reactions after a few minutes from the start of the OHP infusion. Usually, these reactions were seen after 2-17 exposures to OHP (Mean+/ s.e.: 9.4+/-1.07). No patient experienced allergic reactions at his/her first OHP infusion. Eight patients developed a mild reaction consisting of flushing and swelling of the face and hands, itching, sweating and lachrymation. The remaining nine patients showed a moderate-severe reaction with dyspnoea, wheezing, laryngospasm, psycho-motor agitation, tachycardia, precordial pain, diffuse erythema, itching and sweating. Six patients out of 17 were re-exposed to the drug with premedication of steroids and all except one developed the hypersensitivity reaction again. The cumulative dose, the time of exposure to OHP and the clinical features are variable and unpredictable. The risk of developing hypersensitivity reactions in patients treated with a short infusion of OHP cannot be underestimated. PMID- 12888816 TI - Follow-up of patients with Hodgkin's disease following curative treatment: the routine CT scan is of little value. AB - A total of 10-40% of patients with Hodgkin's disease relapse following initial curative therapy. Intensive follow-up is resource intensive and may identify false relapses. We performed a retrospective review of all patients with Hodgkin's disease treated at our centre between 1990 and 1999 to evaluate the utility of the components of follow-up. A total of 107 patients met the inclusion and exclusion criteria. The median age was 33 years and the median duration of follow-up 38 months. The total number of follow-up visits was 1209 and total number of CT scans 283. There were 109 suspected relapses of which 22 proved to be true relapses. Of the latter, 14 were identified clinically, six radiologically and two via lab testing. The routine CT scan detected only two relapses (9%), yet accounted for 29% of the total follow-up costs. Based on data from our centre, the cost per true relapse was $6000 US, 49% incurred by radiological tests. The majority of the cost of follow-up was incurred by routine follow-up (84%) as opposed to the investigation of suspected relapses (16%). We conclude that most true relapses are clinically symptomatic and that the routine CT is an expensive and inefficient mode of routine follow-up. PMID- 12888817 TI - Surgeon workload and survival from breast cancer. AB - The formation of multidisciplinary breast teams across the UK is intended to concentrate the assessment and treatment of breast cancer into the hands of high volume specialists. We undertook a retrospective population-based study in order to determine the trends in surgeon breast cancer workload in Yorkshire, UK, and to investigate whether patients treated by low-workload surgeons had poorer survival. Of 11 329 female breast cancer patients diagnosed in 1989-1994 in Yorkshire, 6% were managed by surgeons with a mean annual workload of less than 10 new patients, while surgeons with workloads of 10-29, 30-49 and >50 treated 21, 21 and 52%, respectively. Over the study period, increasing number of patients were managed by surgeons with higher workloads. Patients treated by low workload surgeons had poorer survival. Five-year survival was 60% in the lowest workload category compared to 68% in the highest category. The relative risk of death was increased by 15% (RR=1.15, 95% CI 1.03-1.28) and by 10% (RR=1.10, 95% CI 1.02-1.18) for patients managed by surgeons with workloads <10 and 10-29 cases per annum in comparison to patients managed by surgeons with workloads of >50. The results of this study suggest increasing site specialisation in breast cancer among general surgeons. It also provides further evidence that the management of patients by surgeons with low workloads decreases overall survival. PMID- 12888818 TI - Effect of the UK government's 2-week target on waiting times in women with breast cancer in southeast England. AB - A government target of a maximum 2-week wait for women referred urgently with suspected breast cancer was introduced in April 1999. We have assessed changes in the distributions of waiting times and the proportions of cases meeting proposed targets before and after this date, using clinical audit data on 5750 women attending 19 hospitals in southeast England during the period July 1997-December 2000, who were subsequently found to have breast cancer. The proportion of cases being seen within 2 weeks of referral rose from 66.0 to 75.2%, and the median wait to first appointment fell from 13.6 to 12.3 days, following the introduction of the government target. The proportion of cases waiting 5 weeks or less between first hospital appointment and treatment fell from 83.8 to 80.3%, and median waits for treatment increased from 21.4 to 24.1 days. We also examined the effects on waiting times of various sociodemographic and care related factors. A total of 85.7% of screening cases vs 67.9% of symptomatic cases were seen within 2 weeks, and 95.0% of cases treated with tamoxifen received treatment within 5 weeks, as opposed to 77.6% of cases treated with surgery, 81.2% of chemotherapy cases and 52.8% of radiotherapy cases. While waiting times from GP referral to first hospital appointment have improved since the introduction of the government target, times from first appointment to treatment have increased, and consequently total waiting times have changed little. PMID- 12888819 TI - Feasibility of quality of life assessment in patients with upper gastrointestinal tract cancer. AB - Quality of life (QOL) is an important outcome after treatment for upper gastrointestinal tract cancer but few studies report good accrual and subsequent attrition is usually high. This study investigated the feasibility of a nurse-led service to obtain longitudinal QOL assessments and explored how clinical and sociodemographic factors influence patients' need for help to complete questionnaires. Fully informed patients were invited into the study. Baseline hospital assessments were scheduled by telephone and thereafter by post unless patients' health indicated the need for a home visit. In all, 128 out of 140 (91%) baseline QOL assessments were performed. Follow-up questionnaire completion was good, with 114 patients (89%) completing all but one of the expected assessments. At baseline, 41 (32%) patients required a lot of help to complete questionnaires. Patients requiring help were more likely to be undergoing palliative treatment than treatment aimed at cure (68 vs 33%; odds ratio 3.48, P<0.01). Patients' with advanced stage cancer of the upper gastrointestinal tract receiving palliative treatment require dedicated staff to ensure good compliance with longitudinal QOL data collection. It is essential to budget for this in clinical trails. PMID- 12888820 TI - Infection with Kaposi's sarcoma-associated herpesvirus (KSHV) and human immunodeficiency virus (HIV) in relation to the risk and clinical presentation of Kaposi's sarcoma in Uganda. AB - A case-control study from Uganda found that the risk of Kaposi's sarcoma increased with increasing titre of antibodies against Kaposi's sarcoma-associated herpesvirus (KSHV) latent nuclear antigens, independently of HIV infection. Clinically, widespread Kaposi's sarcoma was more frequent among patients with HIV infection than in those without, but was not related to anti-KSHV antibody titres. PMID- 12888821 TI - Incidence of malignant neoplasms among HIV-infected persons in Scotland. AB - Among 2574 persons diagnosed with HIV throughout Scotland and observed over the period 1981-1996, cancer incidence compared to the general population was 11 times higher overall; among homosexual/bisexual males, it was 21 times higher and among injecting drug users, haemophiliacs and heterosexuals it was five times higher, mostly due to AIDS-defining neoplasms. However, liver, lung and skin cancers (all non-AIDS-defining) were also significantly increased. PMID- 12888822 TI - Survival from breast cancer among South Asian and non-South Asian women resident in South East England. AB - Ethnic differences in breast cancer survival have been observed in the USA but have not been examined in Britain. We aimed to investigate such differences between South Asian (i.e. those with family roots in the Indian subcontinent) and non-South Asian (essentially British-native) women in England. Primary breast cancer cases incident in 1986 -1993 and resident in South East England were ascertained through the Thames Cancer and Registry and followed up to the end of 1997. Cases of South Asian ethnicity were identified on the basis of their names by using a previously validated computer algorithm. A total of 1037 South Asian and 50 201 non-South Asian breast cancer cases were included in the analysis; 30% of the South Asian (n=312) and 44% (n=22 201) of the non-South Asian cases died during follow-up. South Asian cases had a higher relative survival than non-South Asians throughout the follow-up period. The 10-year relative survival rates were 72.6% (95% confidence interval: 69.0, 75.9%) and 65.2% (64.5, 65.8%) for South Asians and non-South Asians, respectively. The excess mortality rates experienced by South Asians were 82% (72, 94%) of those experienced by non-South Asians (P=0.004). The magnitude of this effect was slightly reduced with adjustment for differences in age at diagnosis, but was strengthened with further adjustment for differences in stage at presentation and socioeconomic deprivation (excess mortality rates in South Asians relative to non-South Asians=72% (63, 82%), P&<0.001). These findings indicate that the higher survival from breast cancer in the first 10 years after diagnosis among South Asian was not due to differences in age at diagnosis, socioeconomic deprivation or disease stage at presentation. PMID- 12888823 TI - Epidemiologic and molecular risk factors for contralateral breast cancer among young women. AB - Women diagnosed with a first breast cancer before the age of 45 years have a greater than 5.0-fold risk of developing a second primary contralateral breast cancer (CBC) than women in the general population have of developing a first breast cancer. Identifying epidemiologic or molecular factors that influence CBC risk could aid in the development of new strategies for the management of these patients. A total of 1285 participants in two case-control studies conducted in Seattle, Washington, who were 21-44 years of age when diagnosed with a first invasive breast carcinoma from 1983 to 1992, were followed through December 2001. Of them, 77 were diagnosed with CBC and 907 tumour tissues from first cancers were analysed. Women with body mass indices (BMIs) >/=30 kg m(-2) had a 2.6-fold greater risk (95% CI: 1.1-5.9) of CBC compared to women with BMIs 80 kg as followed: 0, 3.6% and 4.1% of patients for the Certoparin group and 0, 4.6% and 4.2% of patients for the UFH group. The same was true for major bleeding complications with 0, 2.9% and 1.5% for Certoparin and 0, 3.5% and 4.2% for UFH. Overall mortality was 1.9% in the Certoparin group and 2.7% in the UFH group. Fixed-dose body weight-independent subcutaneous LMWH Certoparin is at least as efficacious and safe as intravenous aPTT-adjusted UFH for the initial treatment of acute proximal DVT. This effect is maintained during a 6-months follow-up of treatment with oral anticoagulation. PMID- 12888873 TI - Comparison of control and stability of oral anticoagulant therapy using acenocoumarol versus phenprocoumon. AB - Variability in the control of oral anticoagulant therapy has been associated with a heightened risk of complications. We compared control of anticoagulation between two commonly used coumarins, phenprocoumon and acenocoumarol, and among anticoagulation clinics. All qualifying patients were managed at six regional anticoagulation clinics in the Netherlands. This retrospective cohort study compiled data during a three-year period from a computerised dosing and management system. Anticoagulation control was expressed as the percent of time within the therapeutic range and stability expressed as the time-weighted variance in the international normalised ratio (INR). Data were available for 22,178 patients of whom 72% were treated with acenocoumarol. INRs of patients who received phenprocoumon were within the therapeutic range 50% of the time compared with 43% for acenocoumarol (OR 1.32, 95% CI 1.24-1.41). Moreover, patients on phenprocoumon required 15% fewer monitoring visits and had more stable INR values. These observations were consistent for all six clinics. There were also sizable differences between the clinics with respect to control and stability of anticoagulation that were stable from year-to-year and were unrelated to the drug used. With its longer half-life of three to five days, phenprocoumon produces more stable anticoagulation than acenocoumarol and should generally be the drug of choice when these are the available choices. The differences observed among clinics suggest that certain clinics employ policies and practices resulting in better control of anticoagulation. PMID- 12888874 TI - A prospective and randomized comparison of the safety and effects of therapeutic levels of enoxaparin versus unfractionated heparin in chronically anticoagulated patients undergoing elective cardiac catheterization. AB - Consecutive patients with atrial fibrillation and/or prosthetic heart valves, receiving chronic anticoagulation with phenprocoumon and scheduled to undergo cardiac catheterization, were randomized to subcutaneous enoxaparin twice daily (n = 32) or intravenous UFH (n = 36). Cardiac catheterization was performed at an international normalized ratio <1.5. Activated partial thromboplastin times and levels of anti-Factor Xa activity were measured daily. The time until effective anticoagulation (primary endpoint) was significantly shorter for enoxaparin than for UFH (1.1 +/- 0.4 days versus 3.7 +/- 2.5 days, p<0.0001). The percentage of days of effective anticoagulation was significantly higher in the enoxaparin group than in the UFH group (93.3 +/- 9.5% versus 53.7 +/- 26.6%, p <0.0001). In conclusion, enoxaparin achieves therapeutic levels of anticoagulation more rapidly and consistently than UFH in chronically anticoagulated patients with prosthetic heart valves and/or atrial fibrillation undergoing cardiac catheterization. PMID- 12888875 TI - Myocardial infarction under the age of 36: prevalence of thrombophilic disorders. AB - It has been suggested that thrombotic tendency increases the risk of myocardial infarction (MI). To investigate the association between the risk of MI at a young age and genetic thrombogenic disorders (G20210A mutation in the prothrombin gene, G1691A mutation in the factor V gene and deficiencies of protein C, protein S and antithrombin III) we conducted a case-control study among 70 survivors of MI who had experienced the event before the age of 36 and 260 healthy subjects. The G20210A mutation in the prothrombin gene was found more often in young patients with MI than among controls (11.4 versus 3.1%). The odds ratio (OR) for MI for carriers versus non-carriers was 4 (95% confidence interval [CI], 1.5 to 11.3). The adjusted OR for major cardiovascular risk factors (smoking, hypecholesterolaemia, diabetes mellitus, hypertension and obesity) was 4.3 (95% CI, 1.3 to 14). The simultaneous presence of both G20210A mutation in the prothrombin gene and smoking further increased the risk of MI compared with nonsmokers and non-carriers (OR, 58; 95% CI, 11.4-294). The G1691A mutation in factor V gene was not associated with an increased relative risk for MI (OR, 0.87; 95% CI, 0.26 to 2.5). Finally, there was no significant difference in the prevalence of deficiencies of protein C, protein S and antithrombin III between cases and controls. In conclusion, our data indicate that the G20210A mutation in the prothrombin gene was the only genetic prothrombotic risk factor associated with the risk of developing MI under the age of 36 years. PMID- 12888876 TI - Replacement therapy with plasma-derived factor VIII concentrates induces skew in T-cell receptor usage and clonal expansion of CD8+ T-cell in HIV-seronegative hemophilia patients. AB - Replacement therapy with factor VIII (FVIII) products causes immune abnormalities in human immunodeficiency virus (HIV)-seronegative hemophilia patients. However, the question remains why an absolute increase in the number of CD8+ T-cells and diminished proliferation responses of lymphocytes to antigen stimulation in vitro occurs in HIV-seronegative hemophilia patients. To examine whether the FVIII products induce skewing of T-cell receptor (TCR) repertoires, TCR variable region alpha-chain and beta-chain repertoires were analyzed for peripheral blood mononuclear cells (PBMCs) from 15 hemophilia patients treated with heated and/or non-heated plasma-derived FVIII concentrates and 10 age-matched healthy adults. Also, T-cell clonality was compared between these groups using complementarity determining region 3 (CDR3) size spectratyping. The skewing of TCR repertoires was significantly greater for hemophilia patients than healthy controls. The extent of T-cell clonality was greater for hemophilia patients than the controls, indicating that clonal T-cells frequently expanded in hemophilia patients. The skew in TCR usage and clonal expansion were primarily observed in patients treated with non-heated plasma-derived products. The spectratyping and sequencing of CDR3 regions revealed that the clonal expansion of T-cells was observed for CD8+ T-cells, but not CD4+ T-cells. These results suggest that extensive expansion of CD8+ T-cells is induced by some viruses other than HIV present in FVIII preparations, and the resulting accumulation of CD8+ T-cells is responsible for changes in peripheral T-cell population in HIV-seronegative hemophilia patients. PMID- 12888877 TI - Intracellular activation of the fibrinolytic cascade in the Quebec Platelet Disorder. AB - The Quebec Platelet Disorder (QPD) is an unusual bleeding disorder associated with increased platelet stores of urokinase-type plasminogen activator (u-PA) and proteolysis of platelet alpha-granule proteins. The increased u-PA and proteolyzed plasminogen in QPD platelets led us to investigate possible contributions of intracellular plasmin generation to QPD alpha-granule proteolysis. ELISA indicated there were normal amounts of plasminogen and plasmin alpha(2)-antiplasmin (PAP) complexes in QPD plasmas. Like normal platelets, QPD platelets contained only a small proportion of the blood plasminogen, however, they contained an increased amount of PAP complexes compared to normal platelets (P < 0.005). The quantities of plasminogen stored in platelets were important to induce QPD-like proteolysis of normal alpha-granule proteins by two chain u-PA (tcu-PA) in vitro. Moreover, adding supplemental plasminogen to QPD, but not to control, platelet lysates, triggered further alpha-granule protein proteolysis to forms that comigrated with plasmin degraded proteins. These data suggest the generation of increased but limiting amounts of plasmin within platelets is involved in producing the unique phenotypic changes to alpha-granule proteins in QPD platelets. The QPD is the only known bleeding disorder associated with chronic, intracellular activation of the fibrinolytic cascade. PMID- 12888878 TI - The role of PAR4 in thrombin-induced thromboxane production in human platelets. AB - There are two protease-activated receptors (PARs), PAR1 and PAR4, in human platelets. It has been suggested that PAR1 mediates platelet responses to low concentrations of thrombin, whereas PAR4 mediates signaling only at high concentrations. In the present study, we used a selective PAR4 blocker, YD-3, to investigate the role of PAR4 in thrombin-induced thromboxane formation in human platelets. YD-3 completely prevented thromboxane production by either a low concentration of thrombin (0.1 U/ml) or the PAR4 agonist peptide GYPGKF. In contrast, YD-3 did not affect thromboxane production caused by the PAR1 agonist peptide SFLLRN, collagen or arachidonic acid. YD-3 also decreased [(3) H]arachidonic acid release from thrombin-stimulated platelets. Moreover, desensitization of platelets with GYPGKF prevented low thrombin-induced thromboxane formation. The decreased thromboxane production by YD-3 is linked to inhibition of calcium influx in thrombin-stimulated platelets. These results suggest that PAR4 plays an important role in the regulation of thromboxane formation in platelets responding to thrombin through prolonged elevation of [Ca(2+)](i) and activation of phospholipase A(2). These data also indicate that PAR4 can be activated by relatively low concentrations of thrombin in human platelets. The selective inhibition of thrombin-induced thromboxane production by YD-3 may be of therapeutic benefit for thrombotic diseases. PMID- 12888879 TI - Affinity and kinetics of P-selectin binding to heparin. AB - P-selectin (CD62P), expressed on stimulated endothelial cells and activated platelets, reacts with P-selectin glycoprotein ligand-1 (PSGL-1, CD162) for leukocyte rolling. It also binds to heparin and heparan sulfate proteoglycans (HSPGs), which attenuates P-selectin mediated adhesions of leukocytes and cancer cells. Here we report that P-selectin mediated adhesion, but not rolling, of the HSPGs bearing human malignant melanoma A375 cells under shear stress. To understand its underlying molecular mechanism, we measured the biophysical properties of this interaction. Heparin inhibited the adhesion of A375 cells to immobilized P-selectin under flow (IC(50) = 3 microM heparin) and neutralized the binding of P-selectin to A375 cells (IC50 = 4 microM heparin). Using surface plasmon resonance technique, we found that P-selectin bound to heparin with a dissociation constant (K(d)) of 115 +/- 6 nM. The measured off rate (k(off)) was 3.15 +/- 0.34 x 10(-3) s(-1) and the calculated on rate (k(on)) was 2.75 x 10(4) M(-1) s(-1). Taken together, our data suggest that the very slow k(off) and the reduced k(on), but apparently not the K(d), are responsible for adhesion, but not rolling of A375 cells, to P-selectin under flow. PMID- 12888880 TI - Enhanced TNF alpha and oxidative stress in patients with heart failure: effect of TNF alpha on platelet O2- production. AB - Experimental studies have suggested that TNF alpha, a pro-inflammatory cytokine, may contribute to the deterioration of cardiovascular function through various mechanisms, including the generation of reactive oxygen species. It has not yet been demonstrated whether TNF alpha has prooxidant activity in patients with heart failure, and what the mechanism eventually resulting in this effect are. We analyzed 42 patients (38 men and 4 women, aged 26 to 74 years) with heart failure, secondary to idiopathic dilated cardiomyopathy (n=21), coronary artery disease (n=15), and valve disease (n=6), and 20 controls (18 men and 2 women, aged 49 to 67 years). Ten patients were in class I, 9 in class II, 15 in class III and 8 in class IV according to NYHA Classification. Blood samples were obtained from each patient to evaluate basal and collagen-induced platelet O(2)( ) production, and plasma TNF alpha. In vivo results showed increased platelet O(2)(-) production and plasma TNF alpha levels in NYHA class III-IV compared with that in controls or in NYHA I-II (p<0,001); platelet O(2)(-) production correlated significantly (R=0,6; p<0,01) with TNF alpha plasma levels. In vitro studies showed TNF alpha dose-dependently (5-40 pg/ml) induced platelet O(2)(-) production, and that this effect was significantly inhibited by its specific inhibitor, WP9QY (1 microM); aspirin (100 microM), AACOCF(3), a specific PLA(2) inhibitor (14 microM), and DPI, an inhibitor of NADPH oxidase, significantly inhibited TNF alpha-mediated platelet O(2)(-) production. This study suggests that in patients with heart failure, enhanced platelet O(2)(-) production is mediated by TNF alpha via activation of arachidonic acid and NADPH oxidase pathways. PMID- 12888881 TI - Endothelial cell function alteration after Junin virus infection. AB - Hematologic involvement is the main feature of Argentine hemorrhagic fever (AHF), an endemo-epidemic disease caused by Junin virus (JV). Since endothelial dysfunction could play a role in AHF-altered hemostasis, we studied human umbilical vein endothelial cell (HUVEC) infection with a virulent (JVv) and a non virulent (JVa) JV strain. Cells were infected by the two JV variants with no detectable apoptosis or cytopathic effect. Both viral variants up-regulated ICAM 1 and VCAM-1 levels, while von Willebrand factor (VWF) production was decreased. Prostacyclin (PGI2) release and decay accelerating factor (DAF) expression were greater in JVv- than in JVa-infected or control cells. Furthermore, nitric oxide (NO) production and endothelial nitric oxide synthase (eNOS) expression was only raised in JVv-infected supernatants. Significant NO and PGI2 values were also detected in AHF patient sera. These data demonstrate that endothelial cell responses are triggered subsequently by JV infection, suggesting that such alterations play a major role in the pathogenesis of AHF and perhaps in other viral-induced hemorrhagic diseases. PMID- 12888882 TI - VEGF-induced HUVEC migration and proliferation are decreased by PDE2 and PDE4 inhibitors. AB - Migration and proliferation of endothelial cells in response to VEGF play an important role in angiogenesis associated to pathologies such as atherosclerosis, diabetes and tumor development. Elevation of cAMP in endothelial cells has been shown to inhibit growth factor-induced proliferation. Our hypothesis was that inactivation of cAMP-specific phosphodiesterases (PDEs) would inhibit angiogenesis. The purpose of this study was to evaluate the effect of PDE inhibitors on in vitro and in vivo angiogenesis, using human umbilical vein endothelial cell (HUVEC) and chick chorioallantoic membrane (CAM) models respectively. Here, we report that: 1) PDE2, PDE3, PDE4 and PDE5 are expressed in HUVEC; 2) EHNA (20 microM), PDE2 selective inhibitor, and RP73401 (10 microM), PDE4 selective inhibitor, are able to increase the intracellular cAMP level in HUVEC; 3) EHNA and RP73401 are able to inhibit proliferation, cell cycle progression and migration of HUVEC stimulated by VEGF; 4) these in vitro effects can be mimic by treating HUVEC with the cAMP analogue, 8-Br-cAMP (600 microM); 5) only the association of EHNA and RP73401 inhibits in vivo angiogenesis, indicating that both migration and proliferation must be inhibited. These data strongly suggest that PDE2 and PDE4 represent new potential therapeutic targets in pathological angiogenesis. PMID- 12888883 TI - Circulating t-PA antigen predicts major adverse coronary events in patients with stable coronary artery disease--a 13-year follow-up. AB - Thrombus formation after rupture of an atherosclerotic plaque plays a crucial role in coronary artery disease (CAD). A decreased endogenous fibrinolytic system and prothrombotic factors are supposed to influence coronary thrombosis. It was our aim to investigate the predictive value of tissue plasminogen activator (t PA) antigen, von Willebrand Factor, Lipoprotein (a) and anti-cardiolipin antibodies for major adverse coronary events in patients with stable CAD in a prospective cohort study of more than 10 years. We observed 141 patients with angiographically proven CAD for a median follow-up period of 13 years. t-PA antigen was the only marker predicting coronary events (logistic regression, p = 0.044) with a poor prognosis for patients in the 5th quintile with an odds ratio of 7.3 (compared to the 1st quintile). The odds ratio even increased to 10.0 for coronary events associated with the "natural course" of CAD excluding events due to restenosis. t-PA antigen had a slightly higher prognostic power (ROC curve; AUC = 0.69) than fasting glucose (AUC = 0.68) and cholesterol (AUC = 0.67). Triglycerides influenced plasma levels of t-PA antigen (regression, p < 0.001). The predictive value of t-PA antigen remained significant after adjustment for inflammation (logistic regression, p = 0.013) and extent of CAD (p = 0.045) but disappeared adjusting for insulin resistance (p = 0.12). In conclusion t-PA antigen predicted coronary events during a very long-term follow-up with a comparable prognostic power to established cardiovascular risk factors. Markers of insulin resistance influenced t-PA antigen and its predictive value. PMID- 12888884 TI - Inhibition of thrombosis in melanoma allografts in mice by endogenous mast cell heparin. AB - An unexplained paradox of malignant melanoma is the apparent failure of the blood within the tumor to clot despite the presence of multiple factors that should promote blood clotting. Here we present histochemical evidence that human and murine melanomas are extensively infiltrated by abundant mast cells. Because mast cells contain the natural anticoagulant heparin, the present studies were aimed at defining the role of mast cell heparin in preventing the blood from clotting within B16 melanoma grafts in C57BL/6 J mice. Mice bearing B16 melanoma grafts were treated with non-specific or specific inhibitors of mast cell heparin (protamine or heparinase, respectively). After the drug treatment there was histologic and functional evidence of selective thrombosis of the blood vessels within the protamine and heparinase treated melanoma grafts. A similar, high degree of thrombosis was also observed in B16 tumors grown in transgenic NDST-2 knockout mice bearing a targeted disruption in the gene coding for mast cell heparin synthesis. The tumors grown in the protamine-treated animals were significantly smaller than the tumors from control (untreated mice). By contrast, the tumors treated with heparinase or grown in the NDST-2 knockout mice were significantly larger than the tumors from control (untreated) mice. We conclude that the intrinsic procoagulant properties of malignant melanoma are neutralized in vivo by the anticoagulant properties of endogenous heparin produced by mast cells that naturally infiltrate the tumor. Our results also suggest that thrombosis and hemostasis within melanoma may play a complex role in modulating the growth of the tumor. PMID- 12888885 TI - Fibrinolysis in organ procurement for transplantation after cardiocirculatory compromise. PMID- 12888886 TI - Elevated plasma von Willebrand factor in a murine model of severe hyperhomocysteinemia. PMID- 12888887 TI - Efficacy and safety of fondaparinux in major orthopedic surgery according to the timing of its first administration. PMID- 12888888 TI - Molecular regulators of angiogenesis in the developing nervous system and adult brain tumors (review). AB - Angiogenesis involves a highly regulated and coordinated interaction of multiple angiogenic factors and is critical for both embryonal development and physiological vessel formation in adults. Angiogenesis is also proven to significantly contribute to the progression of various disease processes. Pathological angiogenesis is thought to recapitulate physiological angiogenesis, however the precise molecular mechanisms are not understood. Various angiogenic genes and gene products have been isolated, purified, and cloned from both neoplastic and normal tissues that contribute to the angiogenic process. In this review we summarize the role of the main angiogenic factors in both normal embryonal vascularization of the embryonal nervous system together with the main brain tumors seen in adults. PMID- 12888889 TI - A commonly deleted region in ovarian cancer on chromosome 19p13.3, not including the OAZ1 gene. AB - Antizyme is a negative regulator of cellular polyamines. The gene for antizyme (OAZ1) is mapped to 19p13.3, where frequent allelic imbalance (AI) is observed in ovarian cancer. The potential role of antizyme 1 in ovarian carcinogenesis was addressed in this study. Mutations of the OAZ1 gene, including the entire coding region and associated promoter region, were examined in 50 primary ovarian tumors and 8 ovarian cancer cell lines by PCR-SSCP and sequencing analyses. A missense mutation in exon 1 and a nucleotide change at the 3'-UTR were detected in an ovarian cancer cell line and its derivative cisplatin resistant cell line. No somatic mutation was detected in primary ovarian tumors, although 7 polymorphic sites were identified. AI of the OAZ1 gene was detected in 7 of 30 informative cases of primary ovarian cancer (23%). Subsequent multiplex fluorescent microsatellite analysis at 7 loci on 19p and at 4 loci on 19q in 50 primary ovarian tumors revealed a commonly deleted region, approximately 4.7 Mb, between the D19S424 and D19S884 loci on 19p13.3 in the vicinity of the OAZ1 locus. The most frequent AI was observed at D19S216 (50%). These results suggest that one or more tumor suppressor genes other than OAZ1 exist near the D19S216 locus on 19p13.3. PMID- 12888890 TI - Abrogation of VEGF expression in human head and neck squamous cell carcinoma decreases angiogenic activity in vitro and in vivo. AB - Angiogenesis is increased in various human cancers, including head and neck squamous cell carcinoma (HNSCC), and correlates with tumor progression and metastasis. Vascular endothelial growth factor (VEGF) has been shown to be a key regulator of angiogenesis. We determined whether VEGF antisense oligonucleotide treatment can decrease angiogenic activity of HNSCC cell lines in vitro and of HNSCC xenografts in vivo. Established human HNSCC cell lines were screened for VEGF expression at both mRNA and protein levels. By using a 21-mer antisense phosphorothioate oligonucleotide targeting the translation start site of human VEGF mRNA, we examined modulation of VEGF expression in cell line supernatants by capture ELISA, and in cell lysates by Western blotting. Human umbilica vein endothelial cells (HUVEC) were grown in conditioned medium produced from the treated tumor cells. Endothelial cell (EC) proliferation was determined by cell count and EC migration was measured using a modified Boyden chamber. Mice with HNSCC xenografts were treated with PBS, VEGF antisense or sense oligonucleotides (10 mg/kg; i.p. injection), respectively and tumor volumes were measured for 5 weeks. VEGF antisense oligonucleotide treatment resulted in a significant reduction of VEGF protein expression compared to sense control. Although the growth rate of the tumor cell lines was not affected, addition of conditioned medium from VEGF antisense-treated tumor cells resulted in decrease of endothelial cell proliferation and migration. VEGF antisense oligonucleotide treatment of HNSCC xenografts resulted in a significant tumor growth suppression. These results suggest that downmodulation of VEGF using antisense oligonucleotides may be a potential therapy for the inhibition of angiogenesis in HNSCC. PMID- 12888891 TI - N-glycosylation is required for the surface localization of MUC17 mucin. AB - The nucleic acid sequence of the human gene, MUC17, indicates that this mucin contains an SEA domain, a transmembrane domain, and putative N-glycosylation sites in the carboxyl terminus. Mucins that possess an SEA domain are usually proteolytically cleaved within that domain to yield two subunits, the smaller of which is associated with the surface membrane. Homogenates of ASPC-1 pancreatic cancer cells showed three main bands of immunoreactivity with alpha-SEA (a polyclonal antibody directed against a site downstream of the postulated cleavage site) after SDS-PAGE and Western blotting (38, 45, and 49 kDa). Experiments utilizing N-glycan specific hydrolases showed that the 38 kDa band contained high mannose glycans whereas the 45 and 49 kDa bands contained complex-type glycans. Only two smaller alpha-SEA reactive bands (30 and 32 kDa) were present after cells had been treated with the N-glycosylation inhibitor tunicamycin. Surface biotinylation studies showed that only the forms possessing complex-type N glycans were localized to the cell surface. Both tunicamycin and brefeldin A, an inhibitor of protein transport, reduced surface localization. In summary, our results indicate that the surface localization of the smaller subunit of MUC17 is dependent on its N-glycosylation status. PMID- 12888892 TI - Angiotensin converting enzyme-independent, local angiotensin II-generation in human pancreatic ductal cancer tissues. AB - Hypovascularity is an outstanding characteristic of pancreatic ductal cancer by diagnostic imaging: most pancreatic ductal cancers are hypovascular or avascular, and tumor vessels are seldom seen on angiography. However, we found that the vasculature was not always poor on angiography of surgically resected specimens of locally advanced pancreatic ductal cancers. To elucidate these controversial findings, we focused on angiotensin II, a vasoconstrictor which is directly produced from angiotensinogen at acidic pH by active trypsin. We examined whether a local angiotensin II-generating system exists in pancreatic ductal cancer tissue. We measured angiotensin II concentration and angiotensin converting enzyme (ACE) activity in tissues from normal pancreas, pancreatic ductal cancers, colon cancers, and hepatocellular carcinomas. After surgically resected specimens were homogenized, angiotensin II concentration and ACE activity in tissues were measured using the florisil method and the Kasahara method, respectively. Tissue angiotensin II levels in pancreatic ductal cancer (n=13) were significantly higher than those of normal pancreas (n=7), colon cancers (n=7), or hepatocellular carcinomas (n=7). However, there was no significant difference in the ACE activity in tissue between them. This study provides in vivo evidence of an ACE-independent, angiotensin II-generating system in pancreatic ductal cancer tissues and suggests that locally formed angiotensin II may act on the pre existing pancreatic arteries around the tumor, leading to formation of hypovascular or avascular regions. PMID- 12888893 TI - Mutational, epigenetic and expressional analyses of caveolin-1 gene in cervical cancers. AB - Caveolin-1 (CAV-1) protein, an integral membrane protein of caveolae membranes, is highly expressed in terminally differentiated cells and down-regulated in cells transformed by human papilloma virus infection. It may also be involved in the tumorigenesis of cervical cancer. CAV-1 gene is regarded as a candidate for the tumor suppressor gene and it can be inactivated in several ways, including point mutations, chromosomal deletions and promoter methylation. We used direct sequencing, methylation specific PCR, and immunohistochemical staining methods to explore the role of CAV-1 gene in the development of cervical cancer. Our results showed that 4 of 72 cases (6%) had methylated CpG-island on the CAV-1 promoter, 17 of 72 cases (26.1%) having no methylation on the promoter showed no expression of CAV-1 protein, and 2 of 72 cases had a GAC right curved arrow GAT transition polymorphism at codon 82. Three types of CAV-1 expression patterns were observed in cervical cancer tissues, and the expression pattern had no relationship with mutation status. From these results, we suggest that CAV-1 gene can be inactivated through mutations, and does not play a role, through methylation of promoter, or an unknown mechanism which may play a role, in the development of cervical cancer. PMID- 12888894 TI - Detection of circulating cancer cells in human oral squamous cell carcinoma. AB - Oral squamous cell carcinoma (OSCC) cells can invade adjacent tissues and the vascular system at an early stage of tumor progression. In the present study, we have attempted to detect circulating cancer cells. We used human OSCC cells expressing green fluorescent protein (GFP) in an orthotopic nude mouse model and were able to detect GFP-expressing cells using a fluorescence activated cell sorter and fluorescence microscopy. We also detected the expression of GFP, squamous cell carcinoma antigen (SCCA), or epidermal growth factor receptor (EGFR) mRNA in the blood of tumor-bearing mice by conventional reverse transcriptase-polymerase chain reaction (RT-PCR) and real-time quantitative RT PCR (TaqMan RT-PCR). TaqMan RT-PCR was the more sensitive method to detect the circulating cancer cells. Furthermore, we examined the expression of SCCA and EGFR mRNA in the peripheral blood of patients with OSCC by conventional RT-PCR and TaqMan RT-PCR. We detected SCCA and EGFR mRNA in few cases by conventional RT PCR, whereas TaqMan RT-PCR showed their expression in about 50% of cases. However, there was no correlation between detection of circulating viable cancer cells and metastasis to lymph nodes and distant organs. These results suggest that TaqMan RT-PCR is a very useful method and both SCCA and EGFR mRNA may be available as a marker for detection of circulating cancer cells. PMID- 12888895 TI - Augmentation of endothelial cell monolayer permeability by hyperthermia but not tumor necrosis factor: evidence for disruption of vascular integrity via VE cadherin down-regulation. AB - Following isolated limb perfusion (ILP) with hyperthermia (H T), TNF and melphalan, there is immediate tumor softening secondary to augmentation of capillary leak in the tumor neovasculature. TNF can induce vascular permeability but is always used with HT during ILP and the contribution of the latter on permeability is not known. This study characterizes the effects of HT and TNF on vascular permeability in vitro. Permeability across confluent human umbilical vein endothelial cells exposed to HT (40 degrees C) with or without 0.1-1000 ng/ml TNF was assessed by quantitating flux of albumin bound Evan's Blue dye from the upper to lower chamber. Immunofluorescent staining for VE-cadherin and F actin was performed after human umbilical vein endothelial cells (hUVECs) were exposed to these conditions. HT induced a significant and reversible increase in permeability compared to untreated hUVECs (p<0.001) whereas barrier function was not altered by TNF. Untreated hUVECs had uniform cell surface staining for VE cadherin, the primary endothelial intercellular adhesion molecule, with colocalization of F-actin cytoskeletal elements. HT resulted in a marked decrease in VE-cadherin staining and contraction of F-actin at sites of endothelial cell cell separation. These data demonstrate that under conditions relevant to those used in ILP, HT but not TNF contributes to a rapid and reversible change in endothelial cell permeability in association with a down regulation of VE cadherin. These data support the use of HT in isolation perfusion and demonstrate a novel mechanism for alterations in microvascular permeability by HT. PMID- 12888896 TI - Identification of signature genes by microarray for acute myeloid leukemia without maturation and acute promyelocytic leukemia with t(15;17)(q22;q12)(PML/RARalpha). AB - Acute myeloid leukemia (AML) has distinct subgroups characterized by different maturation and specific chromosomal translocation. In order to gain insight into the gene expression activities in AML, we carried out a gene expression profiling study with 21 AML samples using cDNA microarrays, focusing on acute promyelocytic leukemia with specific translocation t(15;17)(q22;q12) [French-American-British or FAB-M3 with t(15;17)] and AML without maturation (FAB-M1) characterized by morphologically and phenotypically immature AML blasts and no recurrent chromosomal abnormalities. Using a multivariate sigma-classifier algorithm, we identified 33 strong feature genes that distinguish FAB-M3 with t(15;17) from other AML samples, and 24 strong feature genes that classify FAB-M1. A direct comparison between FAB-M3 with t(15;17) and FAB-M1 led to selection of 13 strong feature genes. Those genes include some known to be related to leukemogenesis and cell differentiation. RIN1, a gene in the ras pathway, was up-regulated in FAB-M3 with t(15;17). Growth factor-binding protein 2 gene was down-regulated in FAB-M1. Huntingtin gene was up-regulated in FAB-M1. Others include syndecan 4, interleukin-2 receptor beta, folate receptor beta, low affinity immunoglobulin gamma, Fc receptor IIC precursor, insulin-like growth factor binding protein 2, and myeloperoxidase, which are involved in cell differentiation. Overexpression of myeloperoxidase in FAB-M3 cells with t(15;17) compared to FAB-M1 cells is consistent with the conventional cytochemical staining pattern. Thus, the study revealed that a morphologically-defined FAB-M1 subtype has a distinct gene expression signature that contributes to its cell differentiation and proliferation as well as FAB-M3 with a recurrent cytogenetic abnormality t(15;17)(q22;q12). PMID- 12888897 TI - Apoptosis induction of vitamin K2 in lung carcinoma cell lines: the possibility of vitamin K2 therapy for lung cancer. AB - Vitamin K2 (menaquinone-4: VK2) has been reported to show apoptosis and differentiation-inducing effects on leukemia cells. Furthermore, the clinical benefits of using VK2 have been demonstrated for the treatment of the patients with acute leukemia and myelodysplastic syndromes. In the present study, we examined the in vitro effects of VK2 on lung carcinoma cell lines LU-139 and LU 130 for small cell carcinomas, PC-14 and CCL-185 for adenocarcinomas, LC-AI and LC-1/sq for squamous cell carcinomas, and IA-LM for large cell carcinoma, respectively. Treatment with VK2 for 48 to 96 h resulted in cell growth suppression in a dose-dependent manner in all cell lines tested. IC50 (50% inhibitory concentration) for VK2 ranged from 7.5 to 75 micro M, and there was no relation between the efficacy of growth suppression by VK2 and tissue type of lung carcinoma cell lines. Morphologic features of the cells treated with VK2 were typical for apoptosis along with caspase-3 activation and becoming positive for APO2.7 monoclonal antibody, an antibody which specifically detects the cell undergoing apoptosis. In addition to the leukemia cell line, LU-139 cells accumulated into G0/G1 phase during 72-h exposure to VK2. Combined treatment of cisplatin plus VK2 resulted in enhanced cytocidal effect compared to the cells treated with either cisplatin or VK2 alone. Since VK2 is a safe medicine without prominent adverse effects including bone marrow suppression, our data strongly suggest the therapeutic possibility of using VK2 for the treatment of patients with lung carcinoma. PMID- 12888898 TI - Protein kinase C isoform expression in ovarian carcinoma correlates with indicators of poor prognosis. AB - In this study expression of protein kinase C alpha (PKCalpha), delta (PKCdelta) and iota (PKCiota) was determined immunohistochemically in formalin-fixed paraffin-embedded tissue specimen of ovarian cystadenomas (n=7), borderline tumours of the ovary (n=8), primary (n=54) and recurrent invasive ovarian carcinomas (n=13). The expression was correlated with clinicopathological parameters and patient survival. In addition, expression of PKCiota was assessed in 3 ovarian carcinoma cell lines (OVCAR-3, SKOV-3, OAW-42) and in one human ovarian surface epithelium (HOSE) cell line. We found expression of PKCalpha in 71.4% of cystadenomas, 50% of borderline tumours, 53.7% of primary and 38.5% of recurrent ovarian carcinomas. PKCdelta was not expressed in epithelium of adenomas, borderline tumours, primary and recurrent ovarian carcinomas. PKCiota was expressed in 51.9% of primary and 46.2% of recurrent ovarian carcinomas but not in cystadenomas and borderline tumours of the ovary. Consistent with these findings ovarian carcinoma cell lines showed strong expression of PKCiota whereas HOSE cells did not. Correlation of PKCalpha and PKCiota expression and clinicopathological features revealed a significant negative correlation of PKCalpha with histopathological grading and a significant positive correlation of PKCiota with histopathological grading and FIGO stage as well as a borderline significant positive correlation with proliferation index. Univariate survival analysis showed that amongst other yet known prognostic parameters (FIGO stage, histopathological grading, proliferation index) PKCiota expression in primary ovarian carcinomas correlated significantly (p=0.024) with a reduced median survival time, but was not an independent prognostic factor. The findings of this study, together with data from functional studies by other groups suggest that alteration of PKC isoform expression may be involved in progression of ovarian carcinomas. PMID- 12888899 TI - Prognostic relevance of MAPK expression in glioblastoma multiforme. AB - The objective of this study was to determine the immunoexpression pattern of the mitogen-activated protein kinase (MAPK), and related signalling proteins [protein kinase C (PKC), phospholipase Cgamma (PLCgamma)], in glioblastoma multi-forme, and to investigate their prognostic value. Paraffin-embedded biopsy samples from 26 patients [13 patients with long-term survival (LTS; N=13; median 28 months, range 13-76 months), and, for comparison, 13 patients with short-term survival (STS; N=13; median 7 months, range 1-12 months)] were investigated for the immunoexpression of MAPK, the activated pMAPK, PKC, PLCgamma, EGFR, and PTEN. Additionally, the MIB-1 proliferation index was determined. The immunoexpression pattern were related to clinical data, including analysis of their prognostic value using the Cox-proportional hazard model. No significant differences were found between STS and LTS in terms of age, Karnofsky performance status, and treatment. Whereas EGFR expression did not differ between STS and LTS and does not influence survival, expression of MAPK and activated pMAPK was significantly correlated with survival time. The percentage of pMAPK expressing cells correlated strongly with the percentage of MIB-1 positive cells. Furthermore, survival in patients with tumors expressing PKC or PLCgamma was significantly shorter. No differences were found for PTEN expression. Our findings indicate that the MAPK pathway is correlated with proliferation in gliomas, and that patient subgroups exist, in which expression of MAPK-related signalling proteins (PKC, PLCgamma) is associated with poorer prognosis. These patient subgroups may benefit from additional chemotherapeutic agents which specifically inhibit these signalling proteins. PMID- 12888900 TI - Effect of formalin fixation on the near-infrared Raman spectroscopy of normal and cancerous human bronchial tissues. AB - Raman spectroscopy is a vibrational spectroscopic technique that can be used to probe molecular changes associated with tissue malignancy. In this report, the effect of formalin fixation on human bronchial tissues was studied by near infrared (NIR) Raman spectroscopy to determine if the variations of Raman spectra caused by formalin fixation would affect the potential diagnostic ability for lung cancer detection. A rapid dispersive-type NIR Raman system with an excitation wavelength of 785 nm was used for this study. Bronchial tissue samples were obtained from six patients with known or suspected malignancies of the lung. Raman spectra of fresh normal and tumor tissue were compared with spectra of formalin-fixed normal and tumor tissue. Changes of the ratios of Raman intensities at 1445 to 1655 cm(-1) and 1302 to 1265 cm(-1) versus formalin fixing times varying from 2 to 24 h were also examined. The major tissue Raman peaks at 1265, 1302, 1445, and 1655 cm(-1) were found in both fresh and fixed bronchial tissues. However, bronchial tissue preserved in formalin showed a progressive decrease in overall intensities of these Raman peaks. Raman contaminations due to formalin were also found in the 980-1100, and 1480-1650 cm(-1) ranges with notable formalin peaks (1041 and 1492 cm(-1)) appearing in the fixed normal and tumor tissues. The results showed that NIR Raman spectra of human bronchial tissues were significantly affected by formalin fixing and tissue hydration. Diagnostic markers at the 980-1100, and 1500-1650 cm(-1) regions derived from fixed tissues do not appear to be applicable for in vivo lung cancer detection. To yield valid Raman diagnostic information for in vivo applications, fresh tissue should be used. If only fixed tissue is available, thorough rinsing of specimens in phosphate-buffered saline (PBS) before spectral measurements may help reduce the formalin fixation artifacts on tissue Raman spectra. PMID- 12888901 TI - Expression and function of activin receptors in human endometrial adenocarcinoma cells. AB - Menstrual cycle-dependent expressions of activin A in normal human endometrial tissues have been reported. Expression of activin receptor mRNAs and increased activin A production were also observed in human endometrial adenocarcinoma tissues, suggesting that activin A might enhance cell proliferation and inhibit apoptotic signaling in endometrial cancer cells. In this study, we have examined the effects of activin A on cell proliferation, anticancer drug-induced apoptosis and Fas-mediated apoptosis in 3 differentiated human endometrial adenocarcinoma cell lines, namely HEC-1, HHUA and Ishikawa. Flow cytometric analyses revealed moderate expressions of all 4 types of activin receptor subunits on the cell surfaces of the 3 cell lines. The proliferations of the 3 endometrial cancer cells were completely unaffected by activin A, whereas it suppressed the cell proliferation of a human ovarian endometrioid adenocarcinoma cell line, OVK-18, in a dose-dependent manner. Moreover, activin A did not affect the apoptotic changes in the 3 endometrial adenocarcinoma cells treated with 4 different anticancer drugs, namely CDDP, paclitaxel, etoposide and SN38. The apoptotic changes in HHUA cells treated with anti-Fas IgM were also unaffected by activin A. These results indicate that the increased activin A production in human endometrial adenocarcinoma tissues in vivo may not stimulate carcinoma cell proliferation or inhibit apoptotic signaling in carcinoma cells. Insensitivity to the usual growth suppression signals induced by activin A might be one of the mechanisms of immortality of human endometrial adenocarcinoma cells. PMID- 12888902 TI - Apoptosis induction and enhancement of cytotoxicity of anticancer drugs by celecoxib, a selective cyclooxygenase-2 inhibitor, in human head and neck carcinoma cell lines. AB - Colorectal carcinomas are well known to highly express COX-2 and their growth is markedly inhibited by COX-2 inhibitors, but little is known about head and neck carcinomas. In this study, we investigated the effect of a selective COX-2 inhibitor, celecoxib, on growth and apoptosis induction of four human head and neck carcinoma cell lines, SCC25, KB, HSG and HSY, in comparison with frequently used COX inhibitor sulindac. Also, we examined whether celecoxib augments the sensitivity of these cell lines to anticancer drugs such as doxorubicin (DOX), vincristine (VCR), cisplatin (CDDP), bleomycin (BLM) and 5-fluorouracil (5-FU). The growth of all cultured cell lines particularly SCC25 and HSG was inhibited by celecoxib and sulindac in a dose-dependent manner. The IC50 of celecoxib was ten times lower than that of sulindac. SCC25 produced ample PGE2 whereas KB, HSG and HSY produced a small amount of PGE2. The PGE2 production and COX-2 expression were inhibited more efficiently by celecoxib than by sulindac. Exogenous addition of PGE2 resulted in an increased cell growth of SCC25 even under the celecoxib treated condition, but not of HSG. These results suggested that PGE2 is involved in the growth of SCC25 but not of HSG. The ability of celecoxib to induce apoptosis is greater than that of sulindac. Treatment of SCC25 and HSG with non cytotoxic 1 micro M or less cytotoxic 5 micro M of celecoxib enhanced the sensitivity of both cell lines to anticancer drugs, particularly in DOX, VCR and BLM two to ten times as demonstrated by lowering of IC50s. The enhanced rate was almost parallel to the degree of apoptosis induction. These findings indicated that a selective COX-2 inhibitor celecoxib inhibits cell proliferation, induces apoptosis and augments sensitivity to anticancer drugs in human head and neck carcinoma cells. Therefore, celecoxib would be useful as biological modulator in treatment of head and neck cancer. PMID- 12888903 TI - Two ways to induce innate immune responses in human PBMCs: paracrine stimulation of IFN-alpha responses by viral protein or dsRNA. AB - In order to study mechanisms of induction of IFN-alpha by Newcastle disease virus (NDV), we used two replicon systems which are based respectively on DNA and RNA of the Semliki forest virus (SFV) and transfected these into baby hamster kidney cells (BHK) which do not produce interferon-alpha. Co-incubation of BHK cells which were transfected with the two vector systems, with human PBMCs, showed that production of IFN-alpha takes place by two different ways. When using the DNA based SFV vector, only transfectants expressing cell surface HN molecules of NDV (and not the mock-transfected cells) elicited such a response via interaction of these HN molecules with viral receptors on PBMCs. In contrast, BHK cells transfected with RNA which had been in vitro transcribed from the RNA-based SFV vector without foreign gene as insert (mock-transfected) elicited a comparable IFN-alpha response. Transfer by transfection of poly(I:C), an analogue of double stranded RNA (dsRNA), into the BHK cells induced also by itself the production of IFN-alpha. Therefore induction of "danger signals" (as double-strand RNA replicative intermediates) might be responsible for this discrepancy observed in IFN-alpha induction in PBMCs between the two studied SFV vector systems based on transfection of DNA and on RNA. These observations highlight two ways of IFN alpha induction which additively may explain the high interferonogenic capacity of NDV as virus: i) via cell-surface expressed HN after transfection of the DNA based SFV replicon and ii) via transfection of self-amplifying RNA. PMID- 12888904 TI - Genome-wide analysis of gene-expression profiles in chronic myeloid leukemia cells using a cDNA microarray. AB - To characterize molecular mechanisms operating in chronic myeloid leukemia (CML) cells with a view toward development of novel therapeutic targets, we analyzed gene-expression profiles of cancer cells from 27 CML patients using a cDNA microarray representing 23,040 human genes. By comparing expression patterns of CML with those of normal cells, we identified 150 genes that were commonly highly up-regulated in CML cells. In addition to 54 genes (34 of them ESTs) whose functions are currently unknown, the up-regulated elements included genes encoding cell-cycle regulators, transcriptional activators, transcriptional factors, and protein kinases as well as proteins already known to be induced in CML, such as some hemoglobins, haptoglobin (HP1), and matrix metalloproteinase 9 (MMP-9), a protein involved in tissue remodeling and tumor invasion. On the other hand, our protocol selected 106 genes, including 13 of unknown function, as being commonly significantly down-regulated in all phases of CML. The results of semiquantitative RT-PCR experiments with 11 representatives of the up-regulated group supported the reliability of our microarray analysis. These data should provide useful information for finding candidate genes whose products might serve as molecular targets for treatment of CML patients. PMID- 12888905 TI - Aberrant methylation in promoter-associated CpG islands of multiple genes in therapy-related leukemia. AB - Methylation profile was analyzed in eleven cases of therapy-related leukemia (t leukemia) for p14, p15, p16, Rb, hMLH1, hMSH2, MGMT, APC, RAR beta, DAPK, RIZ1, FHIT, and SOCS-1 genes by using methylation specific polymerase chain reaction (MSP) analysis. Six (55%) of eleven cases showed methylation of at least one gene. The average time to the development of t-leukemia after the treatment of the primary tumor was significantly shorter in patients with methylation than those without methylation (49.3 months vs. 133.2 months, P=0.044). These results suggest that hypermethylation might be involved in the development of t-leukemia. PMID- 12888906 TI - The pesticide malathion induces alterations in actin cytoskeleton and in cell adhesion of cultured breast carcinoma cells. AB - We have studied the effects of the organophosphorous pesticide malathion on cell viability, actin cytoskeleton, cell adhesion complex E-cadherin/beta-catenin, and Rho and Rac1 GTPases from the human mammary carcinoma cell line MCF-7. Malathion induced cell lethality, determined by the MTT assay, depending on the treatment conditions. Cells incubated with low concentrations of malathion, 16-32 microg/ml, showed high survival rates (>95%) at any evaluated time (1-5 days), whereas complete cell lethality was found using 512 microg/ml and 5 days of treatment. Deep morphological changes were induced with high doses of 64 and 128 microg/ml, and long incubation time (5 days); cells showed perinuclear vacuoles, rounding, shrinkage, and a gradual loss of adhesion. These changes were related to a decrease in the expression of the adhesion molecules, E-cadherin and beta catenin, and to the distribution and reactivity of actin microfilaments to TRITC phalloidin. Disruption of microfilaments, accompanied by the collapse of actin to perinuclear region, were characteristic of cells with loss of adhesion. At lower concentrations, some cells presented deformations on the plasma membrane as lamellipodia-like structures, which were particularly evident from 32 to 128 microg/ml. Conversely, we observed an increase in the expression of Rho and Rac1 GTPases, modulators of actin cytoskeleton and cell adhesion. PMID- 12888907 TI - Anthocyanidins induce apoptosis in human promyelocytic leukemia cells: structure activity relationship and mechanisms involved. AB - Anthocyanidins are the aglycon nucleuses of anthocyanins, which are reddish pigments widely spread in colored fruits and vegetables. To investigate their anti-cancer effect, induction of apoptosis was tested in human promyelocytic leukemia cells (HL-60), which is a valid model for testing antileukemic or general antitumoral compounds. Of six anthocyanidins representing the aglycons of most of anthocyanins, only those with an ortho-dihydroxyphenyl structure on the B ring induce apoptosis, suggesting that the ortho-dihydroxyphenyl structure of anthocyanidins may contribute to the induction of apoptosis. Delphinidin, the most potent inducer, causes apoptosis in a time- and dose-dependent manner. The efficacious induction of apoptosis was observed at 100 micro M for 6 h. Concomitant with the apoptosis, delphinidin stimulates JNK pathway activation including JNK phosphorylation and c-jun gene expression, and activates caspase-3. Antioxidants including N-acetyl-L-cysteine (NAC) and catalase effectively block delphinidin-induced JNK phosphorylation, caspase-3 activation, and DNA fragmentation. Moreover, anthocyanidins directly cause HL-60 cells to generate intracellular hydrogen peroxide. Thus, anthocyanidins may trigger an apoptotic death program through an oxidative stress-involved JNK signaling pathway. The induction of apoptosis by anthocyanins may be the pivotal mechanism by which its chemopreventive action against cancer is based. PMID- 12888908 TI - Anti-Fas antibody-induced apoptosis and its signal transduction in human gastric carcinoma cell lines. AB - The Fas-Fas ligand system is one of the factors involved in cell death signaling. Aberrations in the signaling pathways leading to Fas-mediated apoptosis in tumor cells have been reported in a variety of human malignant tumors. However, the Fas mediated apoptotic pathway has not been sufficiently elucidated in human gastric carcinomas. We examined the apoptotic pathway induced by anti-Fas antibody using seven human gastric carcinoma cell lines. Apoptosis was induced in a delayed fashion and the apoptotic indices (AI) after 48 h were approximately 30-40% in MKN-45 and KATO-III cells, which both showed cleavage of the Bid protein and release of Cytochrome c from the mitochondria. Our data also demonstrated no significant relationship between the expressions of various apoptosis-related proteins and the sensitivity or resistance to anti-Fas antibody-induced apoptosis, as far as we examined. Furthermore, the apoptosis signal was inhibited by treatment with Caspase-9 and -3 inhibitors in MKN-45 and KATO-III. These findings suggest that anti-Fas antibody induced apoptosis through the type II signaling pathway in the human gastric carcinoma cell lines, MKN-45 and KATO-III. PMID- 12888909 TI - Inhibition of NF-kappaB pathway in grape seed extract-induced apoptotic death of human prostate carcinoma DU145 cells. AB - The alarmingly high rate of prostate cancer (PCA) mortality as well as the limited success in the treatment of advanced PCA suggest that additional approaches are needed to control PCA growth and its metastatic potential. A constitutive activation of NF-kappaB family of transcription factors is known to play a major role in chemotherapy resistance in advanced PCA. In recent studies we showed that grape seed extract (GSE) inhibits advanced human PCA growth and induces apoptosis in cell culture and in nude mice. Accordingly, here we assessed the effect of GSE on constitutive and TNFalpha-induced NF-kappaB DNA binding activity and apoptotic death in advanced human prostate carcinoma DU145 cells. Constitutive and TNFalpha-induced NF-kappaB DNA binding activity was inhibited by GSE at doses > or =50 microg/ml and treatments for > or =12 h. This was accompanied by inhibition of IkappaBalpha phosphorylation and IKKalpha kinase activity. A strong induction of apoptosis (P<0.01) was also observed following GSE treatment, while a combination with TNFalpha strongly potentiated apoptosis induction. Our results indicate the potential of developing GSE as an effective cancer therapeutic agent, both alone and in combination with TNFalpha-based chemotherapy of advanced human prostate carcinoma that might prove to be a more effective and less toxic alternative in clinical therapy of PCA. PMID- 12888910 TI - Antiproliferative effect of chlorophyllin derived from a traditional Chinese medicine Bombyx mori excreta on human breast cancer MCF-7 cells. AB - Chlorophyllin (CHL) is the sodium-copper salts of chlorophyll derivatives. In this study, the CHL was derived from a traditional Chinese medicine, the Bombyx mori excreta. The CHL, at concentrations 25-400 microg/ml, reduced the proliferation of HL-60, K-562, S-180, and MCF-7 cells by 8.2-95.7% after 72 h of incubation. The CHL also accumulated G2/M cells and induced apoptosis in the MCF 7 cells. Furthermore, the breast carcinoma cells exhibited lower cyclin D1 and cyclin E levels but higher cyclin B1 level after incubation with the CHL showing that these cyclin changes may be important for the antiproliferative effect of the CHL. PMID- 12888911 TI - Genomic structure and mutational analysis of the human KIF1Balpha gene located at 1p36.2 in neuroblastoma. AB - KIF1a is a member of the kinesin superfamily proteins that are microtubule dependent molecular motors involved in important intracellular functions such as organelle transport and cell division. We previously determined the structure of the human KIF1Bbeta gene, which was found to be a homologue of the murine Kif1bbeta, and demonstrated that the human KIF1Bbeta is a causative gene of Charcot-Marie-Tooth disease type 2A although we did not prove that it is a tumor suppressor gene of neuroblastoma. Here, we identified another isoform of the human KIF1B gene, KIF1Balpha. The KIF1Balpha and KIF1Bbeta are alternative splicing products of the KIF1B gene located on 1p36.2. The KIF1Balpha is distinct from KIF1Bbeta in the C-terminal cargo-binding domain; however, they have the same N-terminal motor domain. We found that the transcript of approximately 7.8 kb of KIF1Balpha was expressed in several tissues, especially in skeletal muscle, by Northern blot analysis. To determine whether this gene is one of the candidate tumor suppressor genes for neuroblastoma (NB) or other pediatric solid tumors, we performed mutational screening of KIF1Balpha in 25 NB, 9 rhabdomyosarcoma, 12 Ewing sarcoma and 24 other pediatric solid tumor cell lines. Using RT-PCR single strand conformation polymorphism analysis and direct sequencing we detected a missense mutation (M807I) in 1 NB cell line (SK-N-SH), 3 silent mutations in 2 NB cell lines and 1 primitive neuroectodermal tumor cell line, respectively. RT-PCR analysis revealed that KIF1Balpha was obviously expressed in almost all of the tumor cell lines examined except NB-1. Furthermore, real-time quantitative RT-PCR showed that there was no significant difference in KIF1Balpha expression between 14 early-stage (stage I and II) and 14 advanced-stage (stage III and IV) NB fresh tumor specimens. These results suggest that KIF1Ba in addition to KIF1Bbeta may not be a candidate tumor suppressor gene for NB. PMID- 12888912 TI - A hypoxic response induced in MatLyLu cells by cobalt chloride results in an enhanced angiogenic response by the chick chorioallantoic membrane. AB - Hypoxia is an important regulator of hypoxia inducible factor-1alpha (HIF-1alpha) and vascular endothelial growth factor (VEGF) expression. Using Dunning rat prostate tumour cells (MatLyLu) we examined the induction of HIF-1alpha and VEGF by hypoxia and if the induction of these proteins would lead to angiogenesis in the chick chorioallantoic membrane (CAM). MatLyLu cells were exposed to 1% O2, in 5.25% CO2 and 94.75% N2, or treated with 100 mM CoCl2 to simulate hypoxia. Conditioned medium was analyzed using Western blots for HIF-1alpha or VEGF. VEGF levels were quantified using ELISA. Hypoxia significantly increased both HIF 1alpha and VEGF protein production. MatLyLu cells (1x10(6) viable cells in 50 microl of medium) grown under normoxic conditions induced in angiogenesis in the CAM, evaluated by the formation of a spoke wheel, and in cross sections by the number of blood vessels, following 5-day culture. This response was significantly increased using CoCl2-treated cells. Culture medium alone with or without CoCl2 was not angiogenic. These results provide direct evidence for the role of hypoxia in the induction of HIF-1alpha and VEGF which act as key angiogenic signals. PMID- 12888913 TI - Overadditive anti-proliferative and pro-apoptotic effects of a combination therapy on colorectal carcinoma cells. AB - The prognosis of advanced colorectal carcinoma (CC) is poor. Established chemotherapy shows only limited efficacy but significant side effects. We investigated how far a combination of tamoxifen (TAM), 9-cis-retinoic acid (CRA) and the fluoroquinolone ciprofloxacin (CIP) synergize to inhibit proliferation and promote apoptosis of CC cells in vitro. The CC cell lines LOVO, CC-531 and SW 403 were incubated with TAM, CRA and CIP (10(minus;4)-10(minus;6) M) as single agents and in combination. Cell proliferation was assessed by bromodeoxyuridin incorporation. Apoptosis was quantified immunohistochemically and by FACS analysis after staining with propidium iodide. Changes in the expression of caspase 3, bax, bcl-2 and p21cip/waf were assessed by quantitative Western blotting. CRA and TAM monotherapy was moderately effective. Their combination enhanced apoptosis from 60% to more than 80% in all cell types. Apoptosis was paralleled by inhibition of proliferation and further potentiated by addition of CIP. The combination effectively up-regulated caspase 3 and bax and down regulated bcl-2 and p21cip/waf. Combinations of biomodulaters act synergistically to inhibit proliferation and promote apoptosis in CC cells. Due to their known safety profile, this justifies clinical trials for colorectal cancer using combinations of these biological response modifiers. PMID- 12888914 TI - Role of cytokeratins, nuclear matrix proteins, Lewis antigen and epidermal growth factor receptor in human bladder tumors. AB - The content of urinary bladder cancer antigen (UBC), tissue polypeptide specific antigen (TPS), nuclear matrix protein 22 (NMP22), and the expression of LewisY carbohydrate antigens and of epidermal growth factor receptor (EGF-R) in bladder tumor tissues were determined. These included 14 well, 6 moderately and 11 poorly differentiated bladder cancers. Cytosol UBC and TPS were higher in the well and in the moderately differentiated bladder tumors than in the poorly differentiated bladder cancers; whereas cytosol NMP22 was higher in the poorly differentiated bladder cancers than in the well and in the moderately differentiated bladder tumors. The Lewis related carbohydrate antigens, evaluated by the reactivity of the tissues to monoclonal antibody B3, were highly expressed in poorly differentiated tumors. The EGF-R was strongly expressed in a large number of poorly differentiated bladder tumors. These data suggest that the determination of cytosol NMP22 and the immunoblotting with B3 and EGF-R antibodies might be useful to obtain more information on the differentiation of bladder tumors. PMID- 12888915 TI - IL-6 is a key factor in growth inhibition of human myeloma cells induced by pravastatin, an HMG-CoA reductase inhibitor. AB - Although recent developments in initial chemotherapeutic regimens and stem cell transplantation have achieved improvements of initial remission for myeloma patients, relapse and recurrence are still major problems. The 3-hydroxy-3 methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors (statins) have been developed for treating hyperlipidemia. Recently, there have been several reports concerning the effects of statins on cancer cells including liver, colon, leukemia, malignant B, stomach, and breast cells. In this study, the in vitro effects of pravastatin on human myeloma cells and the factors closely related to its growth inhibitory effects were examined. Although concentrations were higher than those used clinically, 4 out of 10 myeloma lines showed growth inhibition by pravastatin. The study of factors related to the inhibition indicated IL-6 is important. Indeed, rhIL-6 abolished pravastatin-induced growth inhibition in KMS 21BM cells which did not express IL-6. Statins may be useful in maintenance therapy for myeloma after the screening of IL-6 status. PMID- 12888916 TI - Sialic acids linked to glycoconjugates of Fas regulate the caspase-9-dependent and mitochondria-mediated pathway of Fas-induced apoptosis in Jurkat T cell lymphoma. AB - To clarify the functions of sialic acids linked to glycoconjugates of Fas in Fas induced apoptosis, Jurkat T cells, untreated and treated with neuraminidase, were incubated with anti-Fas monoclonal antibody, CH11. Apoptosis of Jurkat T cells induced by incubation with CH11 was enhanced by the pre-treatment with neuraminidase. By flow cytometry sialylated glycoconjugates were detected on the cell surface of Jurkat T cells using LFA lectin, which specifically reacts with sialic acid, and pre-treatment with Vibrio Cholerae neuraminidase resulted in desialylation of Jurkat cell surface glycoconjugates. The enhancement of Fas induced apoptosis by pre-treatment with neuraminidase was inhibited by z-VAD-fmk, a broad caspase inhibitor, and Ac-LEHD-CHO, an inhibitor of caspase-9, but not by Ac-IETD-CHO an inhibitor of caspase-8 or 6, imipramine, an inhibitor of acidic sphingomyelinase, glutathione, an inhibitor of neutral sphingomyelinase and Fumonisin B1, an inhibitor of ceramide synthase. Mitochondrial membrane potentials (Deltapsim) measured with a Mitocapture assay kit demonstrated that the loss of Deltapsim involved in Fas-induced apoptosis was enhanced by pre treatment with neuraminidase. Furthermore, Western blot analysis using polyclonal antibody (C-20) against Fas detected Fas at about 45 kDa, and pre-treatment with neuraminidase resulted in a reduction of the molecular weight of Fas of about 8 kDa. These data suggest that the enhancement of Fas-induced apoptosis by pre treatment with neuraminidase was mediated by a caspase-9 dependent pathway closely associated with the loss of Deltapsim, not by activation of caspase-8, -6 or acidic and neutral sphingomyelinases, and that sialic acid linked to glycoconjugates of Fas may regulate Fas-induced apoptosis in human T cell lymphoma. PMID- 12888917 TI - Immunization with live HPV-16-transformed mouse cells expressing the herpes simplex thymidine kinase and either GM-CSF or IL-2. AB - From mouse (C57BL/6) HPV-16 transformed cells denoted MK16/1/IIIABC (MK16) a cellular thymidine kinase deficient (cTK-) cell line was isolated. These cTK- cells were transduced by bicistronic recombinant adeno-associated viruses (rAAV) carrying the herpes simplex virus thymidine kinase gene and the gene for either the mouse granulocyte-macrophage colony stimulating factor (GM-CSF) or mouse interleukin-2 (IL-2). Transduced cells were highly sensitive to minute amounts of ganciclovir (GCV) and synthesized moderate amounts of the respective cytokines. A number of cell clones were tested for the cytokine production. The two best producer cell lines, the GM-CSF-producing cells denoted B9 and the IL-2-producing cells denoted 181, were selected for further experiments. Neither B9 nor 181 cells were tumorigenic in syngeneic animals. As inducers of antitumour immunity against challenge with MK16 cells, B9 cells proved superior to the 181 cells. GCV treatment did not markedly influence the level of immunity induced. PMID- 12888918 TI - Inhibition of pre-mRNA splicing by cisplatin and platinum analogs. AB - Our previous study demonstrated that the anticancer agent cis diamminedichloroplatinum (II) (cis-DDP) inhibited the self-splicing activity of the Tetrahymena rRNA. The present study investigated the effects of cis-DDP on pre-mRNA splicing using a HeLa cell nuclear extract. A 2-h exposure of cis-DDP inhibited the splicing of the human B-globin pre-mRNA in a concentration dependent manner. The concentration required for 50% inhibition of splicing (IC50) was 51 microM. Complete inhibition of spliceosome assembly occurred when the extracts were incubated with 150 microM cis-DDP. The inhibition of splicing by cis-DDP occurred at early events during spliceosome formation and to a greater extent if the extract was pre-incubated with cis-DDP in the absence of pre-mRNA. Splicing was inhibited when both pre-mRNA and cis-DDP were added simultaneously to the reaction mixture but not when cis-DDP was added 30 min after splicing was initiated with pre-mRNA. Clinically useful platinum analogs (ormaplatin, carboplatin, cis-tetraplatin and iproplatin) as well as the clinically ineffective Pt(dien)C1+, compound were tested for their ability to inhibit pre mRNA splicing. The Pt(dien)C1+ compound, which acts in a monofunctional manner only, failed to inhibit splicing. A varying degree of splicing inhibition was observed for the other platinum analogs studied; the inhibitory activity decreased in the following order: ormaplatin > cis-tetraplatin > cis-DDP > iproplatin > carboplatin. We describe a novel mechanism that may be involved in the activity and/or toxicity of platinum agents. PMID- 12888919 TI - Hepatic disseminated tumor cells in colorectal cancer UICC stage 4 patients: prognostic implications. AB - Approximately 30-50% of all colorectal cancer patients with a resectable primary tumor will subsequently develop metastatic disease due to tumor cell dissemination. In the case of limited solid hepatic metastasis, resection of the primary tumor and the respective hepatic metastasis can be curative. Our aim was to evaluate the incidence of hepatic DTC in patients with solid liver metastasis and to describe their prognostic impact. Therefore, we applied a sensitive PCR RFLP assay detecting one K-ras mutant among one million wild-type cells. Tumor tissue and liver biopsies from 32 colorectal cancer patients staged UICC 4 undergoing palliative surgery could be obtained intra-operatively and were thereupon screened for the presence of hepatic DTC. The primary tumor of 16 patients (50%) harbored a K-ras mutation and survival of K-ras positive patients was reduced as compared to K-ras negatives (P=0.039). In 8 of the patients (50%) with a K-ras mutated primary, no hepatic DTC were detected, whereas the liver of the remaining 8 patients (50%), showed a coincidence of solid liver metastasis and hepatic DTC. Median survival of patients with solid liver metastasis and hepatic DTC was decreased as compared with patients without any hepatic DTC burden (median survival 165 vs. 240 days; log-rank P=0.951). A subgroup analysis revealed that survival was significantly decreased in the case of bilobal DTC affection as compared with monolobal hepatic DTC affection (median survival 68 vs. 355 days; log-rank P=0.002). We conclude that the detection of hepatic DTC is a powerful prognostic factor. Furthermore, bilobal hepatic DTC involvement might be a contraindication for the resection of solid liver metastasis. PMID- 12888920 TI - 17beta-estradiol-induced activation of ERK1/2 through endogenous androgen receptor-estradiol receptor alpha-Src complex in human prostate cells. AB - We examined the effect of estrogens on mitogen-activated protein kinase (MAPK) in EPN cells, a line of epithelial cells derived from human normal prostate. 17beta estradiol (E2) caused a rapid and transient activation of MAPK (ERK1/2) within 5 min. This effect was counteracted by the anti-estrogen ICI 182-780 and by MEK inhibitor PD098059. The activation of ERK1/2 through 17beta-estradiol triggered simultaneous association of endogenous androgen receptor, estrogen receptor alpha and Src. In addition, E2 stimulated the proliferation of EPN cells, suggesting that the formation of the ternary complex and the consequent activation of ERKs are implicated in the mechanism regulating proliferation of epithelial prostate cells. PMID- 12888921 TI - Akt activity in endometrial cancer cells: regulation of cell survival through cIAP-1. AB - In a number of different cancer including endometrial cancers, tumor suppressor phosphatase tensin homologue (PTEN, a lipid phosphatase) is frequently mutated. PTEN dephosphorylates PI 3-K product, phosphatidylinositol 3,4,5-triphosphate (PIP3), into inactive PIP2 which blocks Akt activation/phosphorylation. In the present study, we have used an endometrial cancer cell line known to possess wild type PTEN (HEC-1-A) and two mutated inactive PTEN protein cell lines (RL-95-2 and Ishikawa) to investigate importance of PI 3-K/PTEN/Akt survival pathway in endometrial cancers. As hypothesised, results showed high levels of Akt1/2 mRNAs and protein phosphorylation in the two mutated PTEN human endometrial cancer cells. To test the possible involvement of Akt in the regulation of survival factors, Bcl-2, XIAP, cIAP-1 and cIAP-2 expression were measured. cIAP-1 protein expression was high in cells expressing phospho-Akt. XIAP and cIAP-2 protein expression was not influenced by the presence of active Akt. Akt phosphorylation decreased and apoptosis was strongly increased in mutated PTEN human endometrial cancer cells in the presence of PI 3-K inhibitor (Wortmannin) which was accompanied by a down-regulation of cIAP-1 protein. Wortmannin had no effect on wild-type PTEN HEC-1-A cell line. Although, Bcl-2 expression was strongly expressed in mutated-PTEN cells, expression remained stable in the presence of Wortmannin suggesting that Bcl-2 is not regulated by Akt. Overexpression of Akt using a constitutively active Akt expression vector resulted in an up-regulation of cIAP-1 expression. These results suggest a pivotal role of Akt in the regulation of endometrial cancer cell survival through the up-regulation of a specific inhibitor of apoptosis protein. PMID- 12888922 TI - Chemoembolization with cisplatin, lipiodol and Gelfoam and subsequent systemic chemotherapy with cisplatin and interferon in patients with hepatocellular carcinoma: a non-randomized prospective study. AB - Arterial chemoembolization with subsequent systemic chemotherapy was assessed prospectively. Of 94 consecutive patients with HCC, 31 patients were considered to have inoperable disease and were selected for chemoembolization. Twenty-two of the 31 patients underwent chemoembolization. In eight patients, technical problems with catheterization prevented the application of therapy, and one patient rejected further treatment. Regimen: Three monthly cycles of chemoembolization with cisplatin 20 mg/m(2) mixed with lipiodol delivered intraarterially with Gelfoam or collagen on day 1, followed by intravenous chemotherapy with cisplatin 60 mg/m(2) on day 2; interferon alpha-2c 30 microg (10 M IU) subcutaneously on days 2, 5, 9, and 12. Three percent of the patients (1/31) (CI 95% 0.08; 16.7) experienced a partial clinical response, in 53% alpha fetoprotein levels decreased by more than 50%. On univariate analysis, performance status, Child score, Okuda stage, albumin levels, and lactate dehydrogenase were found to have an effect on survival. Postchemoembolization syndrome occurred in 68% of the patients, nausea/vomiting grades 3/4 (according to the World Health Organization WHO) in six patients, anemia grade 3 in three patients, leukopenia grade 3 in one patient and thrombocytopenia grade 3 in one patient. This treatment regimen is a very selective procedure. Because of the low response rate it is not recommended for routine clinical use. PMID- 12888923 TI - Inhibition of ErbB-2 and ErbB-3 expression by quercetin prevents transforming growth factor alpha (TGF-alpha)- and epidermal growth factor (EGF)-induced human PC-3 prostate cancer cell proliferation. AB - Because ErbB-2 receptor is involved in hormone-independency for growth and metastasis of prostate cancer cells, the aim was to investigate the effects of quercetin on ErbB-2 and ErbB-3 expression and its critical components such as MAP kinase and PI-3 kinase. Hemocytometric counts and [3H]-thymidine incorporation were used to determine the effects of quercetin, EGF and TGF-alpha on cell proliferation and DNA synthesis in PC-3 and LnCap cells. Changes in ErbB-2, ErbB 3 and components of MAPK and PI-3K pathways were analyzed by Western blot analysis. Treatment of PC-3 and LnCap cells with quercetin resulted in a dose dependent growth inhibition. The rate of DNA synthesis was decreased by 40, 55 and 65% on treatment with 14.5, 29.0 and 58.0 microM of quercetin, respectively. Concomitantly, these treatments led to a dose-dependent decrease in ErbB-2, ErbB 3 and their basal autophosphorylation levels as compared to controls. Cyclin D1 expression and basal phosphorylation of c-Raf, MAPK, Elk-1 and Akt-1 in PC-3 cells was also inhibited by quercetin treatment. Co-treating PC-3 cells with quercetin significantly attenuated EGF- and TGF-alpha-induced growth and phosphorylation of ErbB-2, ErbB-3, c-Raf, MAPK kinase 1/2 (MEK1/2), MAPK, Elk-1 and Akt-1. Since ErbB receptor is important for growth, metastasis and drug resistance, inhibition of ErbB-2 and ErbB-3 by pharmacological doses of quercetin may provide a new approach for treatment of prostate cancers. PMID- 12888924 TI - Identification and characterization of mouse Erbb2 gene in silico. AB - The PPP1R1B-STARD3-TCAP-PNMT-MGC9753-ERBB2-MGC14832-GRB7 locus on human chromosome 17q12 is frequently amplified in human gastric and breast cancer. We have recently identified and characterized human MGC9753 (also known as wild-type CAB2) and mouse Mgc9753. Here, we identified and characterized mouse Erbb2 gene by using bioinformatics. BLAST programs revealed that mouse AK031099 cDNA was derived from mouse Erbb2 gene. Because AK031099 cDNA showed 806 C-->A nucleotide substitution compared with mouse genome draft sequences and mouse Erbb2 ESTs, the nucleotide sequence of mouse Erbb2 cDNA was determined in silico by correcting 806 A of AK031099 cDNA to C. Nucleotide position 48-3818 of mouse Erbb2 cDNA was the coding region. Mouse Erbb2 gene, consisting of 27 exons, was located within the Ppp1r1b-Grb7 locus on the mouse chromosome 11. Mouse Erbb2 protein (1256 aa) showed 87.5% total-amino-acid identity with human ERBB2 protein, and 95.2% total amino-acid identity with rat Erbb2 protein. Mouse Ppp1r1b-Grb7 locus and human Ppp1r1b-Grb7 locus were evolutionarily conserved in the order and the orientation of genes therein. Nucleotide and amino-acid substitution rates of Neurod2 located centromeric to the Ppp1r1b-Grb7 locus were significantly lower than others within the Ppp1r1b-Grb7 locus. This is the first report on the complete coding sequence of mouse Erbb2 gene as well as on the comprehensive comparison of Ppp1r1b-Grb7 locus within the human and mouse genomes. PMID- 12888925 TI - Impact of hemoglobin levels before and during chemotherapy on survival of patients with ovarian cancer. AB - Anemia is a common complication of cancer that has been associated with poor response to treatment and decreased survival in many malignancies. A retrospective chart review was undertaken to determine the effects of hemoglobin (Hb) levels, measured prior to initiation of chemotherapy and before each chemotherapy cycle, and clinical prognostic factors (e.g., age, tumor stage, residual tumor size, hematologic parameters, and type of health insurance) in 250 ovarian cancer patients treated between 1985 and 1998. All patients were scheduled to receive at least 6 courses of systemic chemotherapy. None of the patients received recombinant human erythropoietin. The difference between observed overall survival and its predicted value was computed by multiple regression analysis for each patient with respect to prognostic factors. Hemoglobin levels prior to and during chemotherapy were identified as a prognostic factor for overall survival. Hemoglobin levels > or =12 g/dl were significantly associated with prolonged overall survival (P<0.001). In addition, Hb level correlated with scheduled completion of chemotherapy, overall therapeutic success, tumor stage, age at diagnosis, and residual tumor size (all, P<0.005). Hemoglobin level represents an important prognostic factor for patients with ovarian cancer. This finding supports the use of measures to maintain adequate Hb levels, such as treatment with recombinant human erythropoietin, to improve patient survival. PMID- 12888926 TI - Identification and characterization of ASXL2 gene in silico. AB - Drosophila Asx is a Polycomb group gene. Because Drosophila Asx mutations exhibit anterior and posterior transformations, Drosophila Asx is one of the ETP (Enhancers of trithorax and Polycomb) genes with dual functions in transcriptional activation and silencing. ASXL1 is one of human homologs of Drosophila Asx. Here, we searched for ASXL1-related gene within the human genome by using bioinformatics, and identified the ASXL2 gene. Nucleotide sequence of human ASXL2 cDNA was determined by assembling the nucleotide sequences of human EST AI797346, and partial cDNAs MGC44431 (BC042999) and KIAA1685 (AB051472). Nucleotide sequence of mouse Asxl2 was derived from uncharacterized mouse cDNA 9930017F14 (AK036839). Human ASXL2 (1435 aa) showed 79.4% total-amino-acid identity with mouse Asxl2 (1370 aa), and 29.8% total-amino-acid identity with human ASXL1. ASXN domain (codon 1-86 of ASXL2), ASXM domain (codon 269-380 of ASXL2), and PHD domain (codon 1400-1431 of ASXL2) were conserved between human ASXL2 and ASXL1. Human ASXL2 gene, consisting of at least 13 exons, was mapped to human chromosome 2p23.3, one of recombination hot spots or fragile sites associated with carcinogenesis. The DNMT3A-ASXL2-KIF3C locus on human chromosome 2p23.3 and the DNMT3B-ASXL1-KIF3B locus on human chromosome 20q11.21 were paralogous regions within the human genome. Polycomb group and trithorax group proteins are implicated in embryogenesis and carcinogenesis due to transcriptional regulation of target genes through histone modification and chromatin remodeling. Based on functional conservation and human chromosomal localization, ASXL2 and ASXL1 genes were predicted cancer-associated genes. PMID- 12888928 TI - Malignancy as a solid-phase coagulopathy: implications for the etiology, pathogenesis, and treatment of cancer. AB - This overview of the coagulation biology of cancer is intended primarily for those unfamiliar with the link between the coagulation mechanism and neoplasia. Because the literature on this topic is extensive, citations will be primarily of previous reviews that provide detailed references. The clotting mechanism is fundamentally deranged in malignancy and, unlike the physiological hemostatic response to injury, is incapable of self-attenuation. Clotting activation may predict future cancer and the outcome of existing cancer, and participates in tumor growth. Anticoagulant therapy in patients with malignancy achieves more than control of cancer-related thrombosis. Modification of coagulation reactions relevant to particular tumor types improves the course of the disease. Viewing malignancy as a solid-phase coagulopathy provides insights into possible "upstream" causes of malignant transformation and explanations for the aberrant behavior of transformed cells and suggests innovative experimental interventions. Coagulation biology has broadened and deepened our understanding of neoplasia, and suggested testable strategies for the prevention and treatment of cancer. However, relationships between clotting and cancer progression have been established primarily from model systems. Although limited experience to date in human disease has supported these concepts, resolution of cause-and-effect relationships will require performance of additional rationally designed clinical trials. PMID- 12888929 TI - Cancer-related deep venous thrombosis: clinical importance, treatment challenges, and management strategies. AB - Venous thromboembolic disease (VTE), including deep venous thrombosis (DVT) and pulmonary embolism (PE), is an underdiagnosed and underappreciated clinical problem in cancer patients that results in significant patient morbidity and mortality. Standard treatment practices, including the use of intravenous unfractionated heparin (UFH) for initial anticoagulation, the use of oral warfarin for chronic anticoagulation, and the prescription of only 3 to 6 months total therapy, may not be optimal in the setting of active cancer and ongoing anticancer therapy. Challenges of VTE management in cancer patients include heparin resistance because of excess circulating acute phase proteins, increased recurrence rates during warfarin therapy (target international normalized ratio 2 to 3), limited venous access to support therapeutic monitoring, and increased bleeding rates during anticoagulation. Bleeding during anticoagulation is of particular concern in patients with disease- or chemotherapy-related thrombocytopenia, central nervous system involvement with cancer, and recent surgical intervention. Low-molecular-weight heparins (LMWHs) have been shown to be equally effective and safe for initial anticoagulation compared with UFH and have gained popularity, especially in the setting of VTE in cancer. LMWHs have the advantage of less nonspecific protein binding, subcutaneous weight-based dosing without the need for monitoring in most cases, and less heparin-induced thrombocytopenia. Recent clinical trials have shown LMWHs are at least as effective as oral warfarin for long-term anticoagulation in cancer patients. Interest in LMWHs and several new classes of parenteral and oral anticoagulants extends beyond the primary and secondary prevention of VTE and includes antiangiogenesis, metastasis prevention, and survival prolongation. PMID- 12888930 TI - The statins: multifunctional antithrombotic and antineoplastic drugs. AB - Statins are approved by the Food and Drug Administration (FDA) for the treatment of hypercholesterolemia and have shown remarkable activity in preventing cardiovascular morbidity and mortality. The versatility of statins is increasingly being appreciated, however, and lowering cholesterol is only one attribute among many shared by this class of drugs. Most statins appear to have antithrombotic activity that is unrelated to the ability to reduce cholesterol levels, and several have significant antitumor effects. This article reviews the laboratory and clinical evidence that statins have antithrombotic and anticancer activity, discusses the ways in which these two activities intersect, and proposes novel uses for statins for the treatment of conditions other than hypercholesterolemia. PMID- 12888931 TI - The blood coagulation mechanism in multiple myeloma. AB - Many cancers are associated with hypercoagulability, including multiple myeloma. At least four possible reasons for hypercoagulability have been described in myeloma patients: interference of immunoglobulins on fibrin structure, procoagulant autoantibody production, effects of inflammatory cytokines on endothelium, and acquired activated protein C (APC) resistance. Moreover, injury to endothelium, either by tumor cells or by chemotherapy, may predispose to thrombosis by causing upregulation of adhesion molecules, allowing adhesion of blood cellular elements (platelets, lymphocyte, neutrophils, and tumor cells, which secrete thrombogenic as well as angiogenic substances). In most cases, the pathogenesis of a thrombotic complication in myeloma patients remains unexplained. Administration of chemotherapy may play a larger role in the thrombotic process than a specific abnormality does because thrombotic complications become more prominent after the start of treatment. The recently reported evidence of a non-factor V Leiden APC resistance has increased our understanding of the pathophysiology of this hypercoagulable state. PMID- 12888932 TI - Pathogenesis of increased risk of thrombosis in cancer. AB - Since the observations of Trousseau, not only has the association of cancer and thrombosis been widely recognized but its pathogenesis is now better understood. Attention to the tumor cell as an important source of procoagulants has also contributed to our knowledge of this problem. Tumor cells express tissue factor (TF) and a cancer procoagulant (CP). TF is dormant in the living cell. However, it is activated during apoptosis of the cell, initiating the coagulation cascade and leading to thrombin generation. Because increased apoptosis occurs during treatment with chemotherapeutic agents, hormones, radiation, and hematopoietic growth factors, as well as when there is rapid tumor proliferation, the thrombosis risk is heightened accordingly. These developments have obvious basic and clinical implications. PMID- 12888933 TI - Tissue factor-dependent coagulation activation and impaired fibrinolysis in situ in gastric cancer. AB - Thromboembolism frequently complicates gastric cancer. This study examined the solid phase interaction between gastric cancer and coagulation proteins in situ that may explain coagulation activation and that may contribute to tumor progression and angiogenesis in this tumor type. Immunohistochemical techniques were applied to tissues from 37 cases of adenocarcinoma of the stomach obtained at surgical resection. Fibrinogen was present throughout the tumor stroma. Fibrin and its D-dimer cross-link sites occurred at the host-tumor interface. Subunit "a" of factor (F) XIII and F VII, IX, X, and XII were observed on cancer cells. Prothrombin and prothrombin fragment F1+2 (F1+2) were demonstrated in the tumor stroma on cancer cells and on small blood vessels. Tissue factor (TF) was present on cancer cells and tumor-associated macrophages. Protein C was observed on cancer cells and small blood vessels, whereas protein S was present only in the vascular bed. There was no staining for tissue factor pathway inhibitor (TFPI). High-molecular-weight (HMW) urokinase plasminogen activator (u-PA) antigen was not detected, but weak and inconsistent staining for low-molecular-weight (LMW) u PA was demonstrated on cancer cells. Weak staining for tissue plasminogen activator (t-PA) occurred on cancer cells and in the tumor stroma. In contrast, plasminogen activator inhibitor-1 (PAI-1) expression was strong in the tumor stroma, along with PAI-2 and PAI-3. The endothelium of small stromal blood vessels, particularly near the host-tumor interface, demonstrated von Willebrand factor antigen (vWF Ag). Vascular endothelial growth factor (VEGF) was present on cancer cells and stromal macrophages. These results demonstrate tumor cell associated TF-dependent extravascular coagulation activation in situ in gastric cancer that does not appear to be counterbalanced by TFPI or sufficient fibrinolytic activity. Colocalization of VEGF with hemostatic proteins suggests that they may cooperate in the pathogenesis of gastric cancer. PMID- 12888934 TI - Role of coagulation and fibrinolytic system in prostate cancer. AB - Patients with prostate carcinoma paradoxically have both a hypercoagulable state and a bleeding diathesis. Hypercoagulability manifested by venous and arterial thrombosis has been documented in several large clinical trials. However, many investigators have reported a high risk of postoperative bleeding in prostate cancer patients. Disseminated intravascular coagulopathy has also been commonly noted at different clinical stages of this indolent cancer. In this article we review clinical, laboratory, and experimental evidence for abnormalities of various components of the coagulation and plasminogen pathways and analyze their contribution in prostate cancer growth, progression, and angiogenesis. Finally, we propose potential therapeutic antiangiogenic strategies in patients with prostate cancer. PMID- 12888935 TI - Noninvasive assessment of neoplastic angiogenesis: the role of magnetic resonance imaging. AB - The study of the effect of angiogenesis inhibitors on tumors is limited by our ability to assess their effect in vivo. Approaches that are currently employed have significant limitations. An ideal approach would employ a widely available noninvasive technology that can be used repeatedly to assess the antiangiogenic effect on the same lesions in a serial fashion. We describe here a specialized magnetic resonance imaging (MRI)-based technique that we employ in the study of angiogenesis of brain tumors. This technique, called relative cerebral blood volume (rCBV) mapping, is a noninvasive technique that adds just a few minutes to the conventional MRI study of a human brain tumor in the clinical setting. We hope that such a technique will serve as a model for developing new imaging techniques for the assessment of angiogenesis modulation in other tumor settings. We describe the technical basis and some examples of using rCBV mapping in neoplastic angiogenesis assessment, including a discussion of current limitations and future directions. PMID- 12888936 TI - Occurrence of components of fibrinolytic pathways in situ in laryngeal cancer. AB - Malignancy is characterized by the occurrence of components of coagulation reaction pathways in situ within tumor tissues detectable immunohistochemically. However, tumors vary in the details of this coagulation-cancer interaction. We have previously described tumor cell-associated tissue factor (TF), factor (F) VII, and F X in laryngeal carcinoma tissues. Fibrinogen and F XIIIa were found in the tumor connective tissue. Tissue factor pathway inhibitor (TFPI) occurred in the tumor connective tissue and on microvascular endothelial cells and normal squamous epithelial cells but not in the tumor cells. Fibrin (thrombin-cleaved fibrinogen) existed at the host-tumor interface and the margins of tumor nodules consistent with an active tumor cell-associated clotting pathway in this tumor type. Studies were extended here to detect components of fibrinolytic pathways. Plasminogen and tissue plasminogen activator (t-PA) were detected on laryngeal tumor cells, particularly in more well-differentiated cases. Low-molecular-weight urokinase plasminogen activator (LMW u-PA) was primarily a feature of more undifferentiated laryngeal carcinoma cells. Staining to a lesser extent was found for high-molecular-weight u-PA (HMW u-PA) on tumor cells and various normal cell types in the tumor tissue. Relatively weak and variable tumor cell staining was found for plasminogen activator inhibitors (PAI) 1, 2, and 3. Trace staining was found for u-PA receptor (u-PAR) in differentiated tumor cells. The significance of coagulation and fibrinolytic pathways present in situ to the economy of laryngeal carcinoma remains to be determined. PMID- 12888940 TI - Biomechanical basis of common shoulder problems. AB - Balancing mobility and stability, the biomechanics of the shoulder provides optimal use of the thumb and hand. More than a glenohumeral joint, the shoulder complex consists of four joints and numerous muscles and ligaments. Injuries to the shoulder result from overuse, extremes of motion, and excessive forces. This review describes basic shoulder biomechanics, their role in impingement and instability, and how imaging can detail shoulder function and dysfunction. PMID- 12888941 TI - Elbow: the trochleogingylomoid joint. AB - More complex than a hinged joint, the elbow provides flexion-extension as well as pronation-supination. A unique arrangement of joint surfaces, soft tissue restraints, and muscle forces provides the mobility and stability of this joint. Understanding basic elbow biomechanics will assist the musculoskeletal radiologist in evaluations of this joint. PMID- 12888942 TI - Hip biomechanics: importance to functional imaging. AB - Although well recognized the hip joint is not well understood. In many respects knowledge of the hip is similar to that of the knee several decades ago. Increased understanding of the hip's structure and function will improve our ability to diagnose and treat pre-end-stage hip joint disease. This article describes relevant anatomy and mechanics and details the crucial role that imaging plays in current assessments of hip function. PMID- 12888943 TI - Knee joint biomechanics: relevance to imaging. AB - Over the past 2 decades our understanding of the anatomy and biomechanics of the knee joint has dramatically increased. The translation of this knowledge into clinical practice has greatly improved athletic training and medical treatment when injury occurs. Advances in imaging techniques have played a role in this improvement. The next step for imaging is to move beyond merely providing morphology and to assess the functional mechanics of the knee. This article focuses on the biomechanics of the femorotibial joint and its relevance to imaging. PMID- 12888944 TI - Biomechanics of tendons and tendon failure. AB - Research into the biomechanical properties of tendons had led to a better understanding of the functional and pathophysiologic processes that occur in vivo, particularly in the setting of tendon injury and failure. Increasingly, biomechanical information is being utilized in the clinical setting, guiding patient management in certain circumstances. Differentiating minor and self limiting abnormalities from those in which specific treatment may prevent further disability from altered biomechanics may become an important role for imaging. This article examines normal tendon anatomy and its relationship to tendon biomechanics under both physiological and pathologic conditions. Imaging characteristics of tendons in the physiologic and pathologic states are discussed. We have focused primarily on MRI, as this modality has been studied most intensively and has been shown to offer additional information regarding unsuspected pathology. PMID- 12888945 TI - Foot and ankle biomechanics. AB - With the advent of imaging advancements there has been renewed interest in the foot and ankle. However, many of the basic functions and biomechanical considerations of the bones, joints, and specialized tissues of the normal and dysfunctional foot and ankle remain unfamiliar to many radiologists. This article focuses on the basic biomechanics, normal alignment, and common alignment disorders of the foot and ankle that are relevant to radiologists. PMID- 12888946 TI - A variant of mirror hand: a case report. AB - We report a variant of mirror hand in a 3.5-year-old boy who presented with multiple fingers. The x-ray revealed an attempt at fusion of 2 hypoplastic radii, 1 ulna, and multiple fingers. The unique feature of this case is a broad radius with proximal notching suggesting failed incomplete duplication. The reasons for this duplication and duplication of the fingers appear to be different. PMID- 12888957 TI - Immobilizing enzymes: how to create more suitable biocatalysts. PMID- 12888958 TI - NMR studies of structure and function of biological macromolecules (Nobel lecture). PMID- 12888959 TI - Solvent-free synthesis of rechargeable solid oxygen reservoirs for clean hydrogen oxidation. PMID- 12888960 TI - Plasticity in self-assembly: templating generates functionally different circuits from a single precursor. PMID- 12888961 TI - Efficient methano[70]fullerene/MDMO-PPV bulk heterojunction photovoltaic cells. PMID- 12888962 TI - Transition-metal complexes of an open geodesic polyarene. PMID- 12888963 TI - Rotaxane building blocks bearing blocked isocyanate stoppers: polyrotaxanes through post-assembly chain extension. PMID- 12888964 TI - Asymmetric desymmetrization of meso-1,2-diols by phosphinite derivatives of cinchona alkaloids. PMID- 12888965 TI - First observation of alkali metal ion induced trans-cis isomerization of palladium(II) phosphane complexes containing crown ether moieties. PMID- 12888966 TI - Quantitative correlation of solvent polarity with the alpha-/3(10)-helix equilibrium: a heptapeptide behaves as a solvent-driven molecular spring. PMID- 12888967 TI - Pd-catalyzed nucleophilic alkylation of aliphatic aldehydes with allyl alcohols: allyl, 2-tetrahydrofuryl, and 2-tetrahydropyranyl ethers as useful C3, C4, and C5 sources. PMID- 12888969 TI - Di(isopropylprenyl)borane: A new hydroboration reagent for the synthesis of alkyl and alkenyl boronic acids. PMID- 12888968 TI - Olefin metathesis in the ionic liquid 1-butyl-3-methylimidazolium hexafluorophosphate using a recyclable Ru catalyst: remarkable effect of a designer ionic tag. PMID- 12888970 TI - Highly efficient peptide bond formation to functionalized Wells-Dawson-type polyoxotungstates. PMID- 12888971 TI - Stereospecific total synthesis of the antiviral agent hamigeran B--use of large silyl groups to enforce facial selectivity and to suppress hydrogenolysis. PMID- 12888972 TI - Design and synthesis of paromomycin-related heterocycle-substituted aminoglycoside mimetics based on a mass spectrometry RNA-binding assay. PMID- 12888973 TI - Nickel-catalyzed dimerization and carbosilylation of 1,3-butadienes with chlorosilanes and Grignard reagents. PMID- 12888974 TI - Regioselective hydrosilylation of propargylic alcohols: an aldol surrogate. PMID- 12888975 TI - Synthetic studies on thiostrepton: construction of thiostrepton analogues with the thiazoline-containing macrocycle. PMID- 12888976 TI - Sulfur as the key element for quantitative protein analysis by capillary liquid chromatography coupled to element mass spectrometry. PMID- 12888977 TI - Electrodeposition of nanocrystalline metals and alloys from ionic liquids. PMID- 12888978 TI - Intramolecular C-H activation in alkaline-earth metal complexes. PMID- 12888979 TI - The sudden-polarization effect and its role in the ultrafast photochemistry of ethene. PMID- 12888981 TI - Medical genetic studies in the Amish: historical perspective. PMID- 12888982 TI - Pediatric medicine and the genetic disorders of the Amish and Mennonite people of Pennsylvania. AB - The Clinic for Special Children in Lancaster County, Pennsylvania, is a community supported, nonprofit pediatric medical practice for Amish and Mennonite children who have genetic disorders. Over a 14-year period, 1988-2002, we have encountered 39 heritable disorders among the Amish and 23 among the Mennonites. We emphasize early recognition and long-term medical care of children with genetic conditions. In the clinic laboratory we perform amino acid analyses by high-performance liquid chromatography (HPLC), organic acid analyses by gas chromatography/mass spectrometry (GC/MS), and molecular diagnoses and carrier tests by polymerase chain reaction (PCR) amplification and sequencing or restriction digestion. Regional hospitals and midwives routinely send whole-blood filter paper neonatal screens for tandem mass spectrometry and other modern analytical methods to detect 14 of the metabolic disorders found in these populations as part of the NeoGen Inc. Supplemental Newborn Screening Program (Pittsburgh, PA). Medical care based on disease pathophysiology reduces morbidity, mortality, and costs for the majority of disorders. Among our patients who are homozygous for the same mutation, differences in disease severity are not unusual. Clinical problems typically arise from the interaction of the underlying genetic disorder with common infections, malnutrition, injuries, and immune dysfunction that act through classical pathophysiological disease mechanisms to influence the natural history of disease. PMID- 12888983 TI - Genetic heritage of the Old Order Mennonites of southeastern Pennsylvania. AB - The Old Order Mennonites of southeastern Pennsylvania are a religious isolate with origins in 16th-century Switzerland. The Swiss Mennonites immigrated to Pennsylvania over a 50-year period in the early 18th century. The history of this population in the United States provides insight into the increased incidence of several genetic diseases, most notably maple syrup urine disease (MSUD), Hirschsprung disease (HSCR), and congenital nephrotic syndrome. A comparison between the Old Order Mennonites and the Old Order Amish demonstrates the unique genetic heritage of each group despite a common religious and geographic history. Unexpectedly, several diseases in both groups demonstrate allelic and/or locus heterogeneity. The population genetics of the 1312T --> A BCKDHA gene mutation, which causes classical MSUD, are presented in detail. The incidence of MSUD in the Old Order Mennonites is estimated to be 1/358 births, yielding a corrected carrier frequency of 7.96% and a mutation allele frequency of 4.15%. Analysis of the population demonstrates that repeated cycles of sampling effects, population bottlenecks, and subsequent genetic drift were important in shaping the current allele frequencies. A linkage disequilibrium analysis of 1312T --> A mutation haplotypes is provided and discussed in the context of the known genealogical history of the population. Finally, data from microsatellite marker genotyping within the Old Order Mennonite population are provided that show a significant but modest decrease in genetic diversity and elevated levels of background linkage disequilibrium. PMID- 12888984 TI - Anabaptist genealogy database. AB - In late 1996 we set out to build a computer-searchable genealogy of the Old Order Amish of Lancaster County, Pennsylvania, for use by geneticists. The goals of the project included: 1) using the genealogy to expedite the mapping of genes mutated in three rare recessive disorders under study at the National Institutes of Health (NIH); 2) building a freely available software package, PedHunter, to answer genetically relevant queries on our database and other similar databases; and 3) providing genealogy assistance to researchers outside NIH. All of these scientific goals had to be accomplished while maintaining the confidentiality of the persons in the database and the confidentiality of preliminary research results. We expanded the project to include complementary data sources that contained many individuals who were Anabaptist, but not Amish, and many individuals who never lived in Lancaster County. For this reason, the project was renamed Anabaptist Genealogy Database (AGDB). All of the initial goals of the project have been accomplished, and we recently marked the 5-year anniversary of answering the first of over 100 queries by researchers outside NIH. Thus, it is an opportune time to review the construction of AGDB, summarize its usage to date, and speculate on future projects it might stimulate and facilitate. PMID- 12888985 TI - Type I glutaric aciduria, part 1: natural history of 77 patients. AB - Type I glutaric aciduria (GA1) results from mitochondrial matrix flavoprotein glutaryl-CoA dehydrogenase deficiency and is a cause of acute striatal necrosis in infancy. We present detailed clinical, neuroradiologic, molecular, biochemical, and functional data on 77 patients with GA1 representative of a 14 year clinical experience. Microencephalic macrocephaly at birth is the earliest sign of GA1 and is associated with stretched bridging veins that can be a cause of subdural hematoma and acute retinal hemorrhage. Acute striatal necrosis during infancy is the principal cause of morbidity and mortality and leads to chronic oromotor, gastroesophageal, skeletal, and respiratory complications of dystonia. Injury to the putamen is heralded by abrupt-onset behavioral arrest. Tissue degeneration is stroke-like in pace, radiologic appearance, and irreversibility. It is uniformly symmetric, regionally selective, confined to children under 18 months of age, and occurs almost always during an infectious illness. Our knowledge of disease mechanisms, though incomplete, is sufficient to allow a rational approach to management of encephalopathic crises. Screening of asymptomatic newborns with GA1 followed by thoughtful prospective care reduces the incidence of radiologically and clinically evident basal ganglia injury from approximately 90% to 35%. Uninjured children have good developmental outcomes and thrive within Amish and non-Amish communities. PMID- 12888986 TI - Type I glutaric aciduria, part 2: a model of acute striatal necrosis. AB - Type I glutaric aciduria (GA1) is an inborn error of organic acid metabolism that is associated with acute neurological crises, typically precipitated by an infectious illness. The neurological crisis coincides with swelling, metabolic depression, and necrosis of basal ganglia gray matter, especially the putamina and can be visualized as focal, stroke-like, signal hyperintensity on MRI. Here we focus on the stroke-like nature of striatal necrosis and its similarity to brain injury that occurs in infants after hypoxia-ischemia or systemic intoxication with 3-nitropropionic acid (NPA). These conditions share several features including abrupt onset, preferential effect in the striatum and age specific susceptibility. The pathophysiology of the conditions is reviewed and a model proposed herein. We encourage investigators to test this model in an appropriate experimental system. PMID- 12888987 TI - A study of linkage and association of body mass index in the Old Order Amish. AB - Obesity is thought to have a genetic component with the estimates of heritability ranging from 0.25-0.40. As part of an ongoing study of obesity in the Old Order Amish, seven two- and three-generation families (157 individuals) were assessed for 21 traits related to obesity, including body mass index (BMI) and BMI percentile (a standardized distribution of BMI adjusted for age and sex). Genotyping was performed using a panel of 384 short-tandem repeat markers. In this sample, the estimates of heritability ranged from 0.16-0.31 for BMI and from 0.40-0.52 for BMI-percentile. Model-independent linkage analysis identified candidate regions on chromosomes 1, 5, 7, 8, and 11. Given that several markers on 7q were significant for both BMI and BMI-percentile (P < or = 0.001) and that the structural locus for leptin was located on 7q, this region was considered to be the primary candidate region. Subsequent typing of additional flanking markers on 7q corroborated the original findings. Tests of intrafamilial association for alleles at markers in this candidate region were significant at similar levels. Although there is some evidence for linkage and association in the region containing leptin, there appears to be stronger evidence for linkage (P < or = 0.001) and association (P < or = 0.00001) with BMI in a region 10-15 cM further downstream of leptin, flanked by markers D7S1804 and D7S3070 with peak values from D7S495-D7S1798. Evidence from linkage and association studies suggests that this region (D7S1804-D7S3070) may be responsible, at least in part, for variation in BMI and BMI-percentile in the Old Order Amish. PMID- 12888988 TI - The major mutation in the RMRP gene causing CHH among the Amish is the same as that found in most Finnish cases. AB - Cartilage-hair hypoplasia (CHH), or McKusick type metaphyseal chondrodysplasia, was originally described in the Old Order Amish in the United States and subsequently found to be unusually frequent among Finns. The major mutation causing CHH in Finns is a 70A --> G nucleotide substitution in the RMRP gene, which encodes the untranslated RNA that is a component of mitochondrial RNA processing endoribonuclease. Here we report that the same mutation is the most frequent one, perhaps the only one, in the Amish population in which CHH was first characterized. The fact that the mutation segregates with the same major haplotype in these two populations and others suggests that it is very ancient. Unlike some other ordinarily rare recessive disorders that are limited in their high frequency to a single Amish deme (subisolate), e.g., Ellis-van Creveld syndrome, CHH occurs in high frequency in at least three distinct Amish demes, indicating, along with genealogic data, that there were multiple heterozygotes among the founders, as proposed by McKusick et al. [1965: Bull Johns Hopkins Hosp 116:231-272]. PMID- 12888989 TI - Preparation of macroporous biodegradable poly(L-lactide-co-epsilon-caprolactone) foams and characterization by mercury intrusion porosimetry, image analysis, and impedance spectroscopy. AB - Two poly(L-lactide-co-epsilon-caprolactone) random copolymers containing 5 and 40 mol % of epsilon-CL, namely P(LA-co-CL(5)) and P(LA-co-CL(40)), respectively, have been made macroporous by freeze-drying solutions in dimethylcarbonate. Most of the freeze-dried foams, prepared by varying polymer concentration and cooling rate, exhibited two main pore populations: (1). longitudinally oriented tube-like macropores with diameters >or=100 microm, and (2). interconnected micropores (10 100 microm). Pore characteristics, including macropore density, mean diameter, and interdistance, as well as micropore density, area, and shape, were determined by image analysis of scanning electron micrographs in order to study the influence of processing and formulation parameters on foam structure and properties. The pore orientation and the 3-D texture also were studied by image analysis and impedance spectroscopy. In the case of the P(LA-co-CL(5)), the macropore diameter increased with the cooling rate while the micropore diameter decreased. The micropores also became more circular when the cooling rate was increased. The pore size and morphology of the P(LA-co-CL(40)) were quite unchanged by varying the cooling rate. All the other conditions being the same, the P(LA-co-CL(5)) foams were better organized than the P(LA-co-CL(40)) foams, and pore orientation was improved at the higher cooling rate. Pore size and morphology also can be controlled by changing the polymer concentration (Cp), as we showed by studying P(LA-co-CL(5)) foams prepared by freeze-drying solutions in the 1-10 w/v % Cp range. Macropore density, average diameter, and interdistance of P(LA-co-CL(5)) foams increased with Cp, but the micropore characteristics remained almost unchanged no matter the Cp. The reliability of the characterization methods has been discussed, with special attention to mercury intrusion porosimetry, which is used primarily for measurement of pore volume and pore size distribution. However, this technique is reported here as a destructive and unreliable method for the characterization of fragile P(LA-co-CL(40)) foams. This study shows that image analysis and impedance spectroscopy can give reliable information relative to the pore morphology and anisotropy of freeze-dried foams. PMID- 12888990 TI - Influence of polymeric additives on the biological properties of brushite cements: an experimental study in rabbit. AB - The resorbability and ability of calcium phosphate hydraulic cements to promote new bone formation was investigated in vivo. The effects of two hydrosoluble polymeric additives (hyaluronic acid, and xanthan gum,) on the biological response of two brushite cement formulations (BHC-A vs BHC-B) was investigated. The brushite cements differed in P/Ca (0.71 vs 0.98) and S/Ca (0.10 vs 0.005) atomic ratios and by the presence of calcium sulfate hemihydrate in BHC-A. Polymer-free cements were used as controls. Cement specimens were injected in cylindrical bone defects manually drilled in the distal condyle of rabbit femora. The implants were harvested at 12 and 24 weeks after implantation and subjected to quantitative histomorphometry. The study showed a significantly lower resorption rate for cement BHC-A, which induces the formation of well-mineralized bone in close apposition to the residual material. In contrast, cement BHC-B showed a significant increase of bone formation period and the formation of a thick layer of unmineralized osteoid tissue at the bone/residual cement interface. The presence of xanthan gum made the biological response even worse, particularly in the case of cement BHC-B. The presence of hyaluronic acid has little effect, except for a slight decrease in initial resorption rate, in the case of cement BHC-A. PMID- 12888991 TI - Flexible chitin films as potential wound-dressing materials: wound model studies. AB - Chitin films possessing increased flexibility, softness, transparency, and conformability have been prepared. These attributes enable the potential application of chitin films as occlusive, semipermeable film wound dressings similar to commercial products such as Opsite trade mark. The chitin films are generally nonabsorbent, exhibiting a total weight gain of only up to 120-160% in physiological fluid. Dry chitin films transpire water vapor at a rate of about 600 g/m(2)/24 h, similar to commercial polyurethane-based film dressings, but rises to 2400 g/m(2)/24 h, when wet, which is higher than the water vapor transmission rate of intact skin. The chitin films are nontoxic to human skin fibroblasts, maintaining 70-80% cell viability. Wound studies using a rat model showed no signs of allergenicity or the high inflammatory response associated with biodegradable biomaterials. The chitin films displayed accelerated wound healing properties. Based on histological examination, wound sites dressed with the chitin films stabilized and healed faster, and appeared stronger than those dressed with Opsite trade mark and gauze dressings after 7 days of healing. PMID- 12888992 TI - Long-term craniofacial osteoblast culture on a sodium phosphate and a calcium/sodium phosphate glass. AB - The aim of this study was to determine the characteristics of human craniofacial osteoblasts cultured on sodium phosphate glass and calcium-sodium phosphate glass in a long-term culture of up to 28 days. The characteristics studied were attachment, proliferation, alkaline phosphatase activity, collagen-1 production, and mineralization. A comparison of the degradation rate, measured by mass loss of the glasses, which are intended for use as a component of a novel degradable composite for craniofacial bone repair, was also performed. It was our hypothesis that the glass would be degradable with a change in degradation rate observed by calcium addition and support osteoblast proliferation and expression of the above characteristics. The inclusion of calcium into the reaction mixture significantly decreased the degradation rate, and it is suggested that the slower degradation is the result of pseudo crosslinking (ionic crosslinks rather than covalent bonding) of the polyphosphate chains by the calcium ions. Therefore, twice as many P-O bonds will need to be hydrolyzed for dissolution of the metal phosphate to occur, therefore greatly reducing the rate of hydrolysis. Osteoblasts were able to attach, spread, and proliferate in a manner comparable with the positive control, as shown by analysis of variance. Formation of a collagen-rich mineralized matrix was also observed. The results presented here suggest that a biocompatible soluble glass has been produced, which has potential to be included in a novel biodegradable craniofacial implant. PMID- 12888993 TI - A preliminary study on the enhancement of the osteointegration of a novel synthetic hydroxyapatite scaffold in vivo. AB - Synthetic hydroxyapatite, a bioactive calcium phosphate, is clinically used as a bone replacement bioceramic because of its similarity in composition to bone mineral, biocompatibility, and osteoconductive nature. The aim of this study was to evaluate the bioactivity of a novel synthetic porous hydroxyapatite (PHA) in vivo in rabbit and to investigate the enhancement of its bioactivity and osteointegration. In the investigation reported here, insulin-like growth factor I (IGF-I) has been used to enhance the bioactivity of PHA. Cylindrical PHA implants with or without IGF-I were implanted bilaterally in rabbit femurs. Fluorochrome bone markers were administered at 7-day intervals. The implants with the attached bone were retrieved at postmortem, 1 and 3 weeks after implantation, for histological and histomorphometric analysis. Undecalcified sections stained with toluidine blue showed new bone formation. Mineralization of the new bone formed in the interface, surrounding trabecular bone, and within the pores of the implants was studied. Lamellar bone mineral apposition rate (MAR) was assessed and compared among treatment groups, sham, PHA alone, and PHA with IGF-I (500 ng/implant), by fluorochrome label incorporation using UVL microscopy. We report for the first time, that the supplementation of PHA implants with IGF-I significantly increased new bone formation and MAR (6.58 +/- 0.08 microm/day) compared with implantation of PHA alone (4.08 +/- 0.05 microm/day) or sham operation (3.11 +/- 0.12 microm/day). These results suggest that synthetic PHA might provide a delivery system for bioactive agents to accelerate bone healing in orthopedic procedures. PMID- 12888994 TI - Surface chemistry modulates fibronectin conformation and directs integrin binding and specificity to control cell adhesion. AB - Integrin-mediated cell adhesion to proteins adsorbed onto synthetic surfaces anchors cells and triggers signals that direct cell function. In the case of fibronectin (Fn), adsorption onto substrates of varying properties alters its conformation/structure and its ability to support cell adhesion. In the present study, self-assembled monolayers (SAMs) of alkanethiols on gold were used as model surfaces to investigate the effects of surface chemistry on Fn adsorption, integrin binding, and cell adhesion. SAMs presenting terminal CH(3), OH, COOH, and NH(2) functionalities modulated adsorbed Fn conformation as determined through differences in the binding affinities of monoclonal antibodies raised against the central cell-binding domain (OH > COOH = NH(2) > CH(3)). Binding of alpha(5)beta(1) integrin to adsorbed Fn was controlled by SAM surface chemistry in a manner consistent with antibody binding (OH > COOH = NH(2) > CH(3)), whereas alpha(V) integrin binding followed the trend: COOH >> OH = NH(2) = CH(3), demonstrating alpha(5)beta(1) integrin specificity for Fn adsorbed onto the NH(2) and OH SAMs. Cell adhesion strength to Fn-coated SAMs correlated with alpha(5)beta(1) integrin binding (OH > COOH = NH(2) > CH(3)), and experiments with function-perturbing antibodies demonstrated that this receptor provides the dominant adhesion mechanism in this cell model. This work establishes an experimental framework to analyze adhesive mechanisms controlling cell-surface interactions and provides a general strategy of surface-directed control of adsorbed protein activity to manipulate cell function in biomaterial and biotechnological applications. PMID- 12888995 TI - Piezoelectric and mechanical properties in bovine cornea. AB - The piezoelectric coefficient (d(31)) and Young's modulus (E) were investigated as a function of degree of hydration for bovine cornea. The piezoelectric and mechanical responses observed were anisotropic, and d(31) decreased, whereas E increased with decreasing the degree of hydration. The anisotropic mechanical and electromechanical properties observed seem to be caused by oriented crystalline collagen fibrils. In addition, the loss of water molecules appears to decrease crystallinity (of the collagen) in the cornea. With dehydration of the cornea, a reduction in crystallinity and changes in hydrogen bonding were observed by wide angle X-ray diffraction and Fourier transform infrared measurements. The decrease of piezoelectricity in cornea during dehydration is most likely caused by the increase in modulus and the loss of order to a nonpiezoelectric phase in the collagen. PMID- 12888996 TI - Preparation of antibacterial silver-doped silica glass microspheres. AB - Various types of inorganic substances doped with silver ions have been developed as antibacterial materials, and some have already been commercialized. Colorless and chemically durable materials that slowly release silver ions are, however, still need to be developed. The present authors have previously shown that when a silica glass doped with silver and aluminium ions is prepared using the sol-gel method, the resultant product is colorless, chemically durable, and slowly releases silver ions into water over a long period. The doped silica glass takes a form of microspheres <1 microm in diameter, it is easily mixed with organic polymers, and the mixture can be formed into a thin film or fine fibers, etc. We report on the preparation of silver doped silica glass microspheres having a diameter =1 microm, using the sol-gel method. Initially, tetraethoxysilane was partially prehydrolyzed by water in ethanol, and then aluminium triisopropoxide was added to the solution to form Si-O-Al bonds. Finally, an ammonia solution containing silver nitrate was added to form silica microspheres doped with silver ion together with aluminium ions. The results show monodispersed microspheres 0.4 0.6 microm in diameter were obtained with nominal compositions of Si/Al/Ag = 1/0.01-0.03/0.003-0.03, with a molar ratio of Al/Ag = 1-3.3. The microspheres were colorless, showed a high chemical durability, and slowly released silver ions into water at 37 degrees C. Microspheres with the composition Si/Al/Ag = 1/0.01/0.01 showed excellent antibacterial activity against Escherichia coli. The minimum inhibitory concentration (MIC) of the microspheres was 400, which is less than the MIC value (800) of commercial antibacterial materials. PMID- 12888997 TI - Engineered cellular response to scaffold architecture in a rabbit trephine defect. AB - Tight control of pore architecture in porous scaffolds for bone repair is critical for a fully elucidated tissue response. Solid freeform fabrication (SFF) enables construction of scaffolds with tightly controlled pore architecture. Four types of porous scaffolds were constructed using SFF and evaluated in an 8-mm rabbit trephine defect at 8 and 16 weeks (n = 6): a lactide/glycolide (50:50) copolymer scaffold with 20% w/w tri-calcium phosphate and random porous architecture (Group 1); another identical design made from poly(desaminotyrosyl tyrosine ethyl ester carbonate) [poly(DTE carbonate)], a tyrosine-derived pseudo polyamino acid (Group 2); and two poly(DTE carbonate) scaffolds containing 500 microm pores separated by 500-microm thick walls, one type with solid walls (Group 3), and one type with microporous walls (Group 4). A commercially available coralline scaffold (Interpore) with a 486-microm average pore size and empty defects were used as controls. There was no significant difference in the overall amount of bone ingrowth in any of the devices, as found by radiographic analysis, but patterns of bone formation matched the morphology of the scaffold. These results suggest that controlled scaffold architecture can be superimposed on biomaterial composition to design and construct scaffolds with improved fill time. PMID- 12888998 TI - Performance of degradable composite bone repair products made via three dimensional fabrication techniques. AB - This study analyzed the in vivo performance of composite degradable bone repair products fabricated using the TheriForm process, a solid freeform fabrication (SFF) technique, in a rabbit calvarial defect model at 8 weeks. Scaffolds were composed of polylactic-co-glycolic acid (PLGA) polymer with 20% w/w beta tricalcium phosphate (beta-TCP) ceramic with engineered macroscopic channels, a controlled porosity gradient, and a controlled pore size for promotion of new bone ingrowth. Scaffolds with engineered macroscopic channels and a porosity gradient had higher percentages of new bone area compared to scaffolds without engineered channels. These scaffolds also had higher percentages of new bone area compared to unfilled control defects, suggesting that scaffold material and design combinations could be tailored to facilitate filling of bony defects. This proof-of-concept study demonstrated that channel size, porosity, and pore size can be controlled and used to influence new bone formation and calvarial defect healing. PMID- 12888999 TI - Compatibility in vitro of albumin-heme (O(2) carrier) with blood cell components. AB - Recombinant human serum albumin including 2-[8-[N-(2 methylimidazolyl)]octanoyloxymethyl]-5,10,15,20-tetrakis(alpha,alpha,alpha,alpha o-pivaloylamino)phenylporphinatoiron(II) (albumin-heme; rHSA-FeP) is a synthetic hemoprotein that has sufficient capability to reversibly bind and release O(2) under physiological conditions (pH 7.3, 37 degrees C) similar to hemoglobin and myoglobin. In order to use this albumin-based O(2) carrier as a new class of red blood cell substitutes, its compatibility with blood cell components carefully was investigated in vitro. After the addition of the rHSA-FeP solution into whole blood at 10, 20, and 44 vol %, the FeP concentration in the plasma phase remained constant for 6 h at 37 degrees C in each group, and no significant time dependence was observed in the numbers of red blood cells, white blood cells, or platelets. The microscopic observations clearly showed that the shapes of the red blood cells had not been deformed during the measurement period. With respect to the blood coagulation parameters (prothrombin time and activated partial thromboplastin time), the coexistence of rHSA-FeP had only a negligibly small influence. Also the blood compatibility under dynamic flow conditions was evaluated using a microchannel array flow analyzer. All these results suggest that the albumin-heme has no effect on the morphology of blood cell components in vitro. PMID- 12889000 TI - Multiaxial fatigue behavior of conventional and highly crosslinked UHMWPE during cyclic small punch testing. AB - Previous observations of reduced uniaxial elongation, fracture resistance, and crack propagation resistance of highly crosslinked ultrahigh molecular weight polyethylene (UHMWPE) have contributed to concern that the technology may not be appropriate for systems undergoing cyclic fatigue loading. Using a "total life" approach, we examined the influence of radiation crosslinking on the fatigue response of UHMWPE under cyclic loading via the small punch test. Our goal in this study was to evaluate the suitability of the small punch test for conducting miniature-specimen, cyclic loading, and fatigue experiments of conventional and highly crosslinked UHMWPE. We subjected four types of conventional and highly crosslinked UHMWPE to cyclic loading at 200 N/s and at body temperature in a small punch test apparatus. After failure, the fracture surfaces were characterized with the use of field emission scanning electron microscopy to evaluate the fatigue mechanisms. Cyclic small punch testing under load control was found to be an effective and repeatable method for relative assessment of the fatigue resistance of conventional and highly crosslinked UHMWPE specimens under multiaxial loading conditions. For each of the four conventional and highly crosslinked UHMWPE materials evaluated in this study, fatigue failures were consistently produced according to a power law relationship in the low cycle regimen, corresponding to failures below 10000 cycles. The fatigue failures were all found to be consistent with a single source of initiation and propagation to failure. Our long-term goal in this research is to develop miniature-specimen fatigue testing techniques for characterization of retrieved UHMWPE inserts. PMID- 12889001 TI - Native extracellular matrix coating on Ti surfaces. AB - Osteoblast-like SAOS-2 cells were allowed to synthesize and assemble their extracellular matrix (ECM) on titanium surfaces. After the selective removal of cells, Ti coated with a native ECM was obtained (ECM-Ti). The responses of SAOS-2 cells to ECM-Ti compared with those to peptide sequence RGDS- or fibronectin immobilized Ti were examined, demonstrating the compositional and structural effects needed to trigger the native cell behavior. PMID- 12889002 TI - Ultraviolet light-induced modification of crosslinked hyaluronan gels. AB - Hyaluronan (HA) gels (hylans) crosslinked with divinyl sulfone (DVS) are highly biocompatible and can be structurally modified to obtain desired mechanical properties that are attractive for their use as tissue-engineering scaffolds. However, unmodified hylan gels are not good substrates for cell attachment or infiltration, likely as a result of their smooth surface and the highly anionic nature of HA. This study investigated whether the cell-adhering characteristics of hylan gels could be enhanced by irradiation with ultraviolet (UV) light, with or without prior dehydration. The attachment and proliferation of neonatal rat smooth muscle cells atop these gels was compared with that on unmodified (control; C) or dehydrated (D) gels. UV-induced changes to gel structure and chemistry were characterized by confocal and electron microscopy, and fluorphore assisted carbohydrate electrophoresis (FACE). Cell attachment was sparse on both unmodified (C) and dehydrated (D) gels. Significantly higher levels of cell attachment were observed on the surface of irradiated (UV) and dehydrated irradiated (DUV) gels, likely because of texturing of the gel surface by UV light. In addition, dehydration of gels before UV irradiation created irregular pore-like structures through which cells appeared to migrate into the interior. FACE assays demonstrated that UV-irradiation alters the chemistry of HA, causing limited breakdown of HA chains and DVS crosslinks within gel and possibly creating new crosslinks that have not yet been identified. Because the hylan gels are altered structurally and chemically, binding of cells to the material is likely to be more permanent than possible by other approaches, such as coating of cell-adhesive matrix factors on the gel surface, described previously. The significance of this work is that we have developed a technique for the modification of DVS-crosslinked HA (hylans) to enhance their performance as a cellular scaffold for tissue-engineering applications. PMID- 12889003 TI - Electrostatic spray deposition (ESD) of calcium phosphate coatings. AB - Using electrostatic spray deposition (ESD), thin calcium phosphate layers were deposited onto commercially pure cp-Ti substrates. ESD is a thin-film technique that enables the deposition of inorganic thin films onto metallic substrates using a simple and cheap experimental set-up. The results show that coating structure and morphology can be tailored by choosing the appropriate combination of deposition parameters. Scanning electron microscopy revealed that various surface morphologies, ranging from dense to very porous coatings, can be obtained. Particularly interesting was a unique reticular coating morphology characterized by a three-dimensionally interconnected pore network. X-ray diffraction (XRD) and Fourier transform infrared spectrometry (FTIR) analyses showed that crystalline carbonate apatite coatings were formed after heat treatment of as-deposited ESD coatings. PMID- 12889004 TI - Preparation, characterization, and in vitro degradation of bioresorbable and bioactive composites based on Bioglass-filled polylactide foams. AB - Highly porous poly(D,L-lactide)/Bioglass composites scaffolds were prepared by thermally induced phase separation process of polymer solutions and subsequent solvent sublimation. A series of composite foams with different polymer/Bioglass weight ratios was prepared to study the influence of Bioglass content on the foam characteristics such as porous structure, density, and pore volume. The pore volume was decreased from 9.5 to 5.7 cm(3)/g when the Bioglass content was increased up to 40 wt %, but the overall pore morphology was not affected very much by changing the polymer/glass composition ratio. The composites foams were then incubated in phosphate-buffered saline at 37 degrees C to study the in vitro degradation of the polymer and to detect hydroxyapatite (HA) formation as an indication of their bioactivity. The addition of Bioglass to polymer foams increased the water absorption and weight loss as compared with pure polymer foams. However, the polymer molecular weight, determined by size exclusion chromatography, was found to decrease more rapidly and to a larger extent in absence of Bioglass. This delayed degradation rate in the composite foams was probably caused by the dissolution of alkaline ions from the Bioglass, resulting in a buffering effect of the incubation medium. After incubation for 7 days, HA was detected by X-ray diffractometry and Raman spectroscopy and confirmed by environmental scanning electron microscopy and energy-dispersive X-ray analysis. The porous composites developed here are promising materials for bone regeneration applications because the formation of HA on the surface of the pore walls should provide good environment for the adhesion and proliferation of osteoblasts and osteoprogenitor cells. PMID- 12889005 TI - Development of a novel method for synthesis of a polymeric ultrasound contrast agent. AB - Medical ultrasound is a highly valuable diagnostic tool, especially when compared with other imaging modalities. It is a noninvasive, real-time, portable, extremely safe method compared with X-ray and inexpensive relative to magnetic resonance imaging. However, ultrasound is limited in its ability to distinguish between diseased and normal tissue. This limitation has led to the development of contrast agents. We have produced novel poly (lactic-co-glycolic) acid air-filled microcapsules that work well as ultrasound contrast agents, giving up to 24 and 25 dB enhancement when insonated in the medical imaging range at 5 and 7.5 MHz, respectively. The capsules were fabricated by modifying a double emulsion method to encapsulate camphor in the oil phase and ammonium carbonate in the aqueous phase, and later sublime the encapsulated material, leaving voids capable of being filled with a gas in their place. The role of the surfactant, poly vinyl alcohol, solution temperature, was studied and found to play an important role in the morphology of the capsules, altering their acoustic response. PMID- 12889006 TI - In vivo comparison of the osseointegration of vacuum plasma sprayed titanium- and hydroxyapatite-coated implants. AB - For the last 15 years, orthopedic implants have been coated with hydroxyapatite (HA) to improve implant fixation. The osteoconductive effect of HA coatings has been demonstrated in experimental and clinical studies. However, there are ongoing developments to improve the quality of HA coatings. The objective of this study was to investigate whether a rough and highly crystalline HA coating applied by vacuum plasma spraying (VPS) had a positive effect on the osseointegration of special, high-grade titanium (Ti) implants with the same surface roughness. Ti alloy implants were coated (VPS) with special, high-grade Ti or HA. The osseointegration of the implants was evaluated by either light microscopy or pullout tests after 1, 2, and 4 weeks of unloaded implantation in the cancellous bone of 18 sheep. The interface shear strength increased significantly over all time intervals. By 4 weeks, values had reached approximately 10N/mm(2). However, the difference between the coatings was not significant at any time interval. Direct bone-implant contact was significantly different between the coatings after 2 and 4 weeks, and reached 46% for Ti and 68% for HA implants by 4 weeks. This study indicates that the use of a rough and highly crystalline HA coating, applied by VPS, enhances early osseointegration. Accelerated establishment of secondary implant fixation decreases the risk of early loosening. PMID- 12889007 TI - In vitro bioactivity of silicon-substituted hydroxyapatites. AB - Silicon-containing hydroxyapatites were synthesized by the controlled crystallization method. Chemical analysis, N(2) adsorption, Hg porosimetry, X-ray diffraction, scanning electron microscopy-energy-dispersive X-ray spectroscopy, and X-ray photoelectron spectroscopy (XPS) were used to characterize the hydroxyapatite and to monitor the development of a calcium phosphate layer onto the surface of the substrate immersed in a simulated body fluid, that is, in vitro bioactivity tests. The influence of the silicon content and the nature of the starting calcium and phosphorus sources on the in vitro bioactivity of the resulting materials were studied. A sample of silicocarnotite, whose structure is related to that of hydroxyapatite and contains isolated SiO(4) (4-) anions that isomorphically substitute some PO(4) (3-) anions, was prepared and used as reference material for XPS studies. An increase of the unit cell parameters with the Si content was observed, which indicated that SiO(4) (4-) units are present in lattice positions, replacing some PO(4) (3-) groups. By using XPS it was possible to assess the presence of monomeric SiO(4) (4-) units in the surface of apatite samples containing 0.8 wt % of silicon, regardless the nature of the starting raw materials, either Ca(NO(3))(2)/(NH(4))(2)HPO(4)/Si(OCOCH(3))(4) or Ca(OH)(2)/H(3)PO(4)/Si(OCOCH(3))(4). However, an increase of the silicon content up to 1.6 wt % leads to the polymerization of the silicate species at the surface. This technique shows silicon enrichment at the surface of the three samples. The in vitro bioactivity assays showed that the formation of an apatite like layer onto the surface of silicon-containing substrates is strongly enhanced as compared with pure silicon-free hydroxyapatite. The samples containing monomeric silicate species showed higher in vitro bioactivity than that of silicon-rich sample containing polymeric silicate species. The use of calcium and phosphate salts as precursors lead to materials with higher bioactivity. PMID- 12889008 TI - Heparin inhibits lipopolysaccharide (LPS) binding to leukocytes and LPS-induced cytokine production. AB - The glycosaminoglycan heparin is known to exhibit anti-inflammatory properties unrelated to its anticoagulant activity. However, in a generalized inflammatory response with implanted or extracorporeal devices, the beneficial effect of heparin coating and/or systemic administration is still unclear as well as the precise mechanisms of action. In the present study, we have first studied the effect of heparin on lipopolysaccharide (LPS)-induced cytokine production by human blood monocytes. Our results indicated that the production of interleukin 1alpha, tumor necrosis factor-alpha, and interleukin-8 was significantly decreased when heparin was simultaneously incubated with Escherichia coli LPS. Because the modulation of heparin on monocyte activation could be mediated by its binding via CD14, the main LPS receptor on monocytes, we then studied the binding of LPS and heparin to leukocytes from human blood and to Chinese hamster ovary cells transfected with the human CD14 gene. The data by flow cytometry showed the binding of biotinylated heparin to leukocytes. Moreover, the experiments performed on leukocytes and on CD14-positive Chinese hamster ovary cells indicated that heparin inhibited LPS binding. From our results, we conclude that: 1. heparin is an effective inhibitor of LPS-induced monocyte activation, and 2. heparin inhibits the binding of LPS to cells via a CD14-independent pathway. This study suggests a potentially important therapeutic application for heparin or heparin analogs to prevent inflammation with biomaterials. PMID- 12889009 TI - Prevention of polyurethane valve cusp calcification with covalently attached bisphosphonate diethylamino moieties. AB - OBJECTIVE: Calcification of polyurethane prosthetic valve leaflets causes a major functional impairment. Previously we showed that polyurethane heart valves modified with covalently linked bisphosphonate groups were resistant to calcification in vivo. However, we also found that the highly polar anionic bisphosphonate groups on the polyurethane surface attracted sodium counter ion adsorption, and thereby increased the elastomer's water absorption to 20% of total weight. In this study we address the increased water absorption by investigating the hypothesis that covalently attaching cationic diethylamino groups to the bisphosphonate-modified polyurethane will reduce water absorption. Thus we evaluated the mechanical and in vivo anticalcification properties of heart-valve leaflets composed of this modified polymer. METHODS: Diethylamino and bisphosphonate groups (DBP) were appended to the polyurethane Biospan's hard segment using previously published bromoalkylation methodology. Water absorption and biaxial mechanical and uniaxial failure testing were used to determine the mechanical properties of the DBP-modified polymer. Rat subdermal implants (60 days) and extended (150 days) single pulmonary leaflet replacements in juvenile sheep provided in vivo assessments of the bisphosphonate-modified polyurethane. RESULTS: The water absorption properties of the DBP-modified polymers and unmodified polyurethanes were 1.86 and 2.3 %, respectively. Biaxial mechanical tests showed the DBP-modified polymer was more compliant than the unmodified control material, but all polymeric material had similar uniaxial failure properties. In both rat subdermal and sheep circulatory implants, the DBP modified polyurethane resisted calcification, as assessed by scanning electron microscopy, with complete calcification inhibition in prosthetic sheep valve leaflet replacements. CONCLUSION: DBP polyurethane possesses physical (water absorption) and biomechanical properties comparable to unmodified polyurethane and can resist intrinsic heart-valve leaflet calcification in blood-stream implants. PMID- 12889010 TI - Reactive oxygen species inhibited by titanium oxide coatings. AB - Titanium is a successful biomaterial that possesses good biocompatibility. It is covered by a surface layer of titanium dioxide, and this oxide may play a critical role in inhibiting reactive oxygen species, such as peroxynitrite, produced during the inflammatory response. In the present study, titanium dioxide was coated onto silicone substrates by radio-frequency sputtering. Silicone coating with titanium dioxide enhanced the breakdown of peroxynitrite by 79%. At physiologic pH, the peroxynitrite donor 3-morpholinosydnonimine-N-ethylcarbamide (SIN-1) was used to nitrate 4-hydroxyphenylacetic acid (4-HPA) to form 4-hydroxy 3-nitrophenyl acetic acid (NHPA). Titanium dioxide-coated silicone inhibited the nitration of 4-HPA by 61% compared to aluminum oxide-coated silicone and 55% compared to uncoated silicone. J774A.1 mouse macrophages were plated on oxide coated silicone and polystyrene and stimulated to produce superoxide and interleukin-6. Superoxide production was measured by the chemiluminescent reaction with 2-methyl-6-[p-methoxyphenyl]-3,7-dihydroimidazo[1,2-a]pyrazin-3-one (MCLA). Titanium dioxide-coated silicone exhibited a 55% decrease in superoxide compared to uncoated silicone and a 165% decrease in superoxide compared to uncoated polystyrene. Titanium dioxide-coated silicone inhibited IL-6 production by 77% compared to uncoated silicone. These results show that the anti inflammatory properties of titanium dioxide can be transferred to the surfaces of silicone substrates. PMID- 12889011 TI - Exposure of fibrinogen-adherent platelets to plasma proteins: a new method for studying protein interactions with platelets. AB - To understand the effects of mediators in coagulating blood at biomaterial surfaces, it is important to use methods that resemble the normal sequence of events in wound healing around implants. The initial adhesion of platelets from whole blood onto material surfaces is mediated by the fibrinogen receptor glycoprotein IIb/IIIa, as shown in a previous study (Broberg et al., J Lab Clin Med 2002; 139:163-172). In this study, isolated platelets were adhered to fibrinogen and exposed to IgG, von Willebrand factor, or thrombin. The response was detected as the number of adherent platelets, the spreading of platelets, the exposure of CD62P (P-selectin), and the release of platelet factor 4 (PF4), ADP, and ATP. These results were compared to the response of platelets adhering to surfaces coated with the same proteins. Fibrinogen-adherent platelets exposed to thrombin generated the significantly highest exposure of CD62P and release of PF4, ADP, and ATP. When platelets were adhered to different protein coatings, von Willebrand factor generated the most CD62P exposure, IgG generated the most PF4 release, and thrombin generated the highest concentration of ADP. These results indicate that protein interactions with platelets may generate different results, depending on the mode of protein exposure. PMID- 12889012 TI - Electrochemical preparation of chitosan/hydroxyapatite composite coatings on titanium substrates. AB - Composite coatings containing brushite (CaHPO(4). 2H(2)O) and chitosan were prepared by electrochemical deposition. The brushite/chitosan composites were converted to hydroxyapatite/chitosan composites in aqueous solutions of sodium hydroxide. The coatings ranged from approximately 1 to 15% chitosan by weight. Qualitative assessment of the coatings showed adhesion significantly improved over that observed for electrodeposited coatings of pure HA. PMID- 12889013 TI - Tailored delivery of active keratinocyte growth factor from biodegradable polymer formulations. AB - We report the results of a high throughput screening campaign that is aimed to develop a biodegradable polymer-based formulation to deliver active keratinocyte growth factor (KGF) and provide a means to tune the KGF delivery rate. A statistical design strategy was used to prepare and screen a series of polymer blends that were composed of poly(lactic acid) (PLA), poly(glycolic acid) (PGA), and the surfactant sodium bis(ethylhexyl)sulfosuccinate (Aerosol-OT, AOT). Chloroform was the solvent. Our high throughput screening method used a two tiered assessment strategy. At Level 1, we identified "lead" KFG-loaded formulations that exhibited KGF emission spectra that were the most similar to the native KGF spectrum recorded in buffer. At Level 2, we used steady-state emission and a homogeneous polarization immunoassay strategy to determine the concentration of total and active KGF, respectively, liberated from the lead formulations during biodegradation. After preparing and screening 2500 formulations, we identified several viable, lead formulations. An analysis of the data showed that the combination of PLA, PGA, and AOT were important to yield a high fraction of active KGF upon release from the formulation; no combination of any two together produced an effect as good as the ternary formulation. The optimum formulations that yielded the highest fraction of active KGF upon release had the following general features: PLA/PGA (w/w) near unity, AOT loading of 100 200 mM, water/AOT mole ratio of 10-20, and a pH between 6 and 8. PLA alone cast from chloroform delivered KGF, but that KGF did not bind to anti-KGF antibodies (i.e., it was inactive). We can tune the KGF release kinetics by more than two orders of magnitude while maintaining the KGF activity upon liberation from the formulation by adjusting the PLA molecular weight. PMID- 12889014 TI - Seeding cells into needled felt scaffolds for tissue engineering applications. AB - Tissue engineering methods are under development that will enable the repair or replacement of a variety of tissues, including articular cartilage and bone. To engineer functional tissue it is necessary that scaffolds initially be seeded with a large number of cells distributed evenly throughout the scaffold structure. It previously has been shown that, compared to static seeding conditions, seeding scaffolds under dynamic conditions facilitates high seeding densities and even distributions of cells (Li et al., Biotechnology Progress 2001;17:935-944). The efficiency of seeding HOSTE85 cells and bovine chondrocytes into needled felt scaffolds following agitation at different speeds was determined. Seeding efficiency was determined using the Hoechst 33258 assay, and cell viability was assessed using the Alamar Blue trade mark assay. The distribution of cells within the scaffolds was imaged using scanning electron microscopy. It was found that the optimum seeding conditions varied for HOSTE85 cells and bovine chondrocytes, with different agitation speeds leading to different seeding efficiencies, cell viabilities, and distributions of cells within scaffolds. The optimum agitation speeds for seeding a high number of viable cells into scaffolds so that they were arranged evenly were 300 rpm for HOSTE85 cells and 200 rpm for bovine chondrocytes. PMID- 12889015 TI - Resistance of Lactobacillus casei in plastic-composite-support biofilm reactors during liquid membrane extraction and optimization of the lactic acid extraction system. AB - Lactic acid fermentations were performed with plastic-composite-support (PCS) disks in solvent-saturated media with Lactobacillus casei subsp. rhamnosus (ATCC 11443). The PCS disks contained 50% (w/w) polypropylene, 35% (w/w) ground soybean hulls, 5% (w/w) yeast extract, 5% (w/w) soybean flour, and 5% (w/w) bovine albumin. Bioassays were performed by growing L. casei in solvent-saturated media after soaking the PCS disks. Eighteen different solvent and carrier combinations were evaluated. Overall, L. casei biofilm fermentation demonstrated the same lactic acid production in solvent-saturated medium as suspended cells in medium without solvents (control). To evaluate PCS solvent-detoxifying properties, two bioassays were developed. When solvent-saturated medium in consecutive equal volumes (10 mL then 10 mL) was exposed to PCS, both media demonstrated lactic acid fermentation equal to the control. However, when solvent-saturated medium with two consecutive unequal volumes (10 mL then 90 mL) was exposed to PCS, some degree of toxicity was observed. Furthermore, iso-octane, tributylphosphate (TBP), and Span 80 were optimized for recovery as 91%, 5%, and 4% (v/v), respectively, with a 1:1 ratio of 1.2 M Na(2)CO(3) stripping solution. Also, recovery by emulsion liquid extraction in the hollow-fiber contactor was minimal due to low recovery at pH 5.0 and incompatibility of the solvent and hollow-fiber material. These results suggest that PCS biofilm reactors can benefit lactic acid fermentation by eliminating the toxic effect from solvent leakage into the fermentation medium from liquid-liquid extractive integrated fermentations. PMID- 12889016 TI - Clonal expansion of adult rat hepatic stem cell lines by suppression of asymmetric cell kinetics (SACK). AB - Adult stem cells have potential use for several biomedical applications, including cell replacement therapy, gene therapy, and tissue engineering. However, such applications have been limited due to difficulties encountered in expanding functional adult stem cells. We have developed a new approach to the problem of adult stem cell expansion based on the suppression of asymmetric cell kinetics (SACK). We postulated that asymmetric cell kinetics, required for adult stem cell function, were a major barrier to their expansion in culture. As such, conversion of adult stem cells from asymmetric cell kinetics to symmetric cell kinetics would promote their exponential expansion and longterm propagation in culture. The purine nucleoside xanthosine (Xs), which promotes guanine ribonucleotide biosynthesis, can be used to reversibly convert cells from asymmetric cell kinetics to symmetric cell kinetics. We used Xs supplementation to derive clonal epithelial cell lines from adult rat liver that have properties of adult hepatic stem cells. The properties of two Xs-derived cell lines, Lig-8 and Lig-13, are described in detail and compared to properties of adult rat hepatic cell lines derived without Xs supplementation. The Xs-derived cell lines exhibit Xs-dependent asymmetric cell kinetics and Xs-dependent expression of mature hepatic differentiation markers. Interestingly, Lig-8 cells produce progeny with properties consistent with hepatocyte differentiation, while Lig-13 progeny cells have properties consistent with bile duct epithelium differentiation. A stable adult cholangiocyte stem cell line has not been previously described. Consistent with the principles of their derivation, the SACK-derived hepatic cell lines exhibit neither senescence nor tumorigenic properties, and their differentiation properties are stable after longterm culture. These characteristics of SACK-derived stem cell lines underscore asymmetric cell kinetics as an essential adult stem cell property with potential to be the basis for a general approach to expansion and propagation of diverse adult stem cells. PMID- 12889017 TI - Elevated temperature effects on the oxidant/antioxidant balance in submerged batch cultures of the filamentous fungus Aspergillus niger B1-D. AB - In the present study the relationship between oxidative stress and elevated culture temperature was examined in an industrially relevant fungal culture, Aspergillus niger B1-D. For the first time, both the intracellular levels of the main stressor species (superoxide radical [O(2) (.-)]) and activities of cellular defensive enzymes (superoxide dismutase [SOD], catalase [CAT], and glutathione peroxide [GPx]) were quantified at varying temperature (25, 30, 35, 40 degrees C) to more fully characterize culture response in different growth phases. Elevated culture temperature led to increased O(2) (.-) levels in various culture phases. In the exponential phase this was due to an enhanced generation of O(2) (.-), whereas in stationary phase a decreased dismutation rate may also have contributed. CAT activities generally increased with culture temperature, whereas GPx activity changed little as temperature rose, indicating that GPx played only a minor role in destroying H(2)O(2) in this A. niger. The combination of elevated temperature (35 degrees C) and increased O(2) supply (50% enrichment) led to decreased levels of O(2) (.-) compared to the cultivation at 35 degrees C gassed with air, probably due to enhanced activity of the alternative fungal respiratory pathway. Our findings indicate that while elevated cultivation temperature does clearly induce oxidative stress events, mechanistically, it does so by a rather more complex route than previous studies indicate. Elevated temperature caused a marked disparity in the activities of SOD and CAT, very distinct from the integrated increase in activity of these enzymes in response to oxidative stress. PMID- 12889018 TI - Three-dimensional numerical approach to investigate the substrate transport and conversion in an immobilized enzyme reactor. AB - This numerical study evaluates the momentum and mass transfer in an immobilized enzyme reactor. The simulation is based on the solution of the three-dimensional Navier-Stokes equation and a scalar transport equation with a sink term for the transport and the conversion of substrate to product. The reactor consists of a container filled with 20 spherical enzyme carriers. Each of these carriers is covered with an active enzyme layer where the conversion takes place. To account for the biochemical activity, the sink term in the scalar transport equation is represented by a standard Michaelis-Menten approach. The simulation gives detailed information of the local substrate and product concentrations with respect to external and internal transport limitations. A major focus is set on the influence of the substrate transport velocity on the catalytic process. For reactor performance analysis the overall and the local transport processes are described by a complete set of dimensionless variables. The interaction between substrate concentration, velocity, and efficiency of the process can be studied with the help of these variables. The effect of different substrate inflow concentrations on the process can be seen in relation to velocity variations. The flow field characterization of the system makes it possible to understand fluid mechanical properties and its importance to transport processes. The distribution of fluid motion through the void volume has different properties in different parts of the reactor. This phenomenon has strong effects on the arrangement of significantly different mass transport areas as well as on process effectiveness. With the given data it is also possible to detect zones of high, low, and latent enzymatic activity and to determine whether the conversion is limited due to mass transfer or reaction resistances. PMID- 12889019 TI - Modeling chromate reduction in Shewanella oneidensis MR-1: development of a novel dual-enzyme kinetic model. AB - Chromate (Cr(VI)) reduction tests were performed with nitrate- and fumarate-grown stationary phase cultures of Shewanella oneidensis MR-1 (henceforth referred to as MR-1) and disappearance of Cr(VI) was monitored over time. A rapid initial decrease in Cr(VI) concentration was observed, which was followed by a slower, steady decrease. These observations appear to be consistent with our previous results indicating that Cr(VI) reduction in MR-1 involves at least two mechanisms (Viamajala et al., 2002b). Modeling of metal reduction kinetics is often based on single-enzyme Michaelis-Menten equations. However, these models are often developed using initial rates and do not always match actual reduction profiles. Based on the hypothesis that multiple Cr(VI) reduction mechanisms exist in MR-1, a model was developed to describe the kinetics of Cr(VI) reduction by two parallel mechanisms: (1) a rapid Cr(VI) reduction mechanism that was deactivated (or depleted) quickly, and (2) a slower mechanism that had a constant activity and was sustainable for a longer duration. Kinetic parameters were estimated by fitting experimental data, and model fits were found to correspond very closely to quantitative observations of Cr(VI) reduction by MR-1. PMID- 12889020 TI - Substrate inhibition kinetics for toluene and benzene degrading pure cultures and a method for collection and analysis of respirometric data for strongly inhibited cultures. AB - We present an evaluation of the qualitative and quantitative effects that high concentrations of benzene and toluene have on the growth rate of several pure cultures that use these compounds as their sole carbon and energy source. The cultures employed were five widely studied environmental isolates: Pseudomonas putida F1, P. putida mt2, P. mendocina KR, Ralstonia pickettii PKO1, and Burkholderia cepacia G4. Three cultures degraded toluene following a pattern consistent with the kinetic model of Wayman and Tseng (1976) while the other two followed a modification of this model introduced by Alagappan and Cowan (2001). The pattern followed for benzene degradation was different than that for toluene degradation for all four capable pure cultures and consistent with that described by the model of Luong (1987). Mechanisms of substrate inhibition and solvent toxicity are discussed, used to conceptually evaluate the reasons for the differences in inhibition behavior, and used to support a call for more widespread use of the empirical, terminal substrate concentration inhibition models employed here. We also present the methodology developed to overcome a limitation commonly encountered when attempting to collect oxygen uptake data for use in quantifying substrate inhibition kinetics. The experimental method was effective for use in the collection of high quality data and the substrate inhibition models most useful in representing the growth of bacteria on these solvents are those that show a complete loss of activity at high concentration rather than the more popular asymptotic inhibition models. PMID- 12889021 TI - Artificial regulatory networks and cascades for discrete multilevel transgene control in mammalian cells. AB - Prototype drug-adjustable heterologous transcription control systems designed for gene therapy applications typically show sigmoid dose-response characteristics and enable fine-tuning of therapeutic transgenes only within a narrow inducer concentration range of a few nanograms. However, the design of clinical dosing regimes which achieve tissue-specific concentrations with nanogram precision is yet a "mission impossible." Therefore, most of today's transcription control systems operate as ON/OFF switches and not in a true adjustable mode. The availability of robust transcription control configurations which lock expression of a single therapeutic transgene at desired levels in response to fixed clinical doses of different inducers rather than minute concentration changes of a single inducer would be highly desirable. Based on in silico predictions, we have constructed a variety of mammalian artificial regulatory networks by interconnecting the tetracycline- (TET(OFF)), streptogramin- (PIP(OFF)), and macrolide- (E(OFF)) repressible gene regulation systems as linear (auto)regulatory cascades. These networks enable multilevel expression control of several transgenes in response to different antibiotics or allow titration of a single transgene to four discrete expression levels by clinical dosing of a single antibiotic: 1) high expression in the absence of any antibiotic (+++), 2) medium level expression following addition of tetracycline (++), 3) low level expression in response to the macrolide erythromycin (+), and 4) complete repression by streptogramins such as pristinamycin (-). The first-generation artificial regulatory networks exemplify modular interconnections of different heterologous gene regulations systems to achieve multigene expression, fine tuning, or to design novel control networks with unprecedented transgene regulation properties. Such higher-level transcription control modalities will lead the way towards composite artificial regulatory networks able to effect complex therapeutic interventions in future gene therapy and tissue engineering scenarios. PMID- 12889022 TI - Evaluation of effective diffusion coefficient and intrinsic kinetic parameters on azo dye biodegradation using PVA-immobilized cell beads. AB - An immobilized mixed culture (Aeromonas hydrophila, Comamonas testosteroni, and Acinetobacter baumannii) was prepared by entrapment into phosphorylated polyvinyl alcohol (PVA) gel beads. The unsteady-state diffusion mechanism in a gel bead was applied to estimate the effective diffusion coefficients (D(e)) and the partition coefficients (K(p)) of azo dye. In addition, a simple method was developed to determine the intrinsic kinetic parameters of immobilized cells from observed reaction rates and the intrinsic kinetic parameters were then verified by fitting the experimental data into the reaction-diffusion model in a batch reactor running at a well-stirred state. The calculated effectiveness factor (eta(cal)) approached unity at Thiele modulus (Phi) < 0.3 (i.e., d(p) < 0.475 mm). The experimental effectiveness factor (eta(exp)) was in the range of 0.71-0.45 for a corresponding sphere diameter (d(p)) of 1.91 +/- 0.16 to 4.43 +/- 0.07 mm at an initial dye concentration of 200 mg/L. The results show that intraparticle diffusion resistance has a significant effect on the azo dye biodegradation rate. PMID- 12889023 TI - Acetohydroxyacid synthase: a new enzyme for chiral synthesis of R phenylacetylcarbinol. AB - We have found that acetohydroxyacid synthase (AHAS) is an efficient catalyst for the enantiospecific (> or =98% enantiomeric excess) synthesis of (R) phenylacetylcarbinol (R-PAC) from pyruvate and benzaldehyde, despite the fact that its normal physiological role is synthesis of (S)-acetohydroxyacids from pyruvate and a second ketoacid. (R)-phenylacetylcarbinol is the precursor of important drugs having alpha and beta adrenergic properties, such as L-ephedrine, pseudoephedrine, and norephedrin. It is currently produced by whole-cell fermentations, but the use of the isolated enzyme pyruvate decarboxylase (PDC) for this purpose is the subject of active research and development efforts. Some of the AHAS isozymes of Escherichia coli have important advantages compared to PDC, including negligible acetaldehyde formation and high conversion of substrates (both pyruvate and benzaldehyde) to PAC. Acetohydroxyacid synthase isozyme I is particularly efficient. The reaction is not limited to condensation of pyruvate with benzaldehyde and other aromatic aldehydes may be used. PMID- 12889024 TI - Down-regulation of acetate pathway through antisense strategy in Escherichia coli: improved foreign protein production. AB - A problem with the use of Escherichia coli to produce foreign proteins is that although endogenously produced acetate is physiologically indispensable, it inhibits protein expression. Here we firstly employed an antisense RNA strategy as an elaborate metabolic engineering tool to partially block biosynthesis of two major acetate pathway enzymes, phosphotransacetylase (PTA) and acetate kinase (ACK). Three recombinant plasmids containing antisense genes targeting either or both of pta and ackA were constructed, and their effects on the acetate pathway and foreign protein productivity compared to control plasmid without any antisense genes were determined in E. coli BL21. Green fluorescent protein (GFP) was employed as a model foreign protein, and timing of antisense expression was controlled by using the intrinsic ackA promoter. We found that the antisense method partially reduced mRNA levels of target enzyme genes and, over time, lowered the concentration of acetate in culture media in all antisense-regulated strains. Notably, total production of GFP was enhanced 1.6- to 2.1-fold in antisense-regulated strains, even though the degree of acetate reduction was not significantly large. It was revealed that the acetate pathway has more critical roles in cellular physiology than expected in the previous reports. When the scale of culture was increased, enhancement of protein production became larger, demonstrating that this antisense strategy can be successfully applied to practical large-scale protein production processes. PMID- 12889025 TI - In silico prediction and validation of the importance of the Entner-Doudoroff pathway in poly(3-hydroxybutyrate) production by metabolically engineered Escherichia coli. AB - The metabolic network of Escherichia coli was constructed and was used to simulate the distribution of metabolic fluxes in wild-type E. coli and recombinant E. coli producing poly(3-hydroxybutyrate) [P(3HB)]. The flux of acetyl-CoA into the tricarboxylic acid (TCA) cycle, which competes with the P(3HB) biosynthesis pathway, decreased significantly during P(3HB) production. It was notable to find from in silico analysis that the Entner-Doudoroff (ED) pathway flux increased significantly under P(3HB)-accumulating conditions. To prove the role of ED pathway on P(3HB) production, a mutant E. coli strain, KEDA, which is defective in the activity of 2-keto-3-deoxy-6-phosphogluconate aldolase (Eda), was examined as a host strain for the production of P(3HB) by transforming it with pJC4, a plasmid containing the Alcaligenes latus P(3HB) biosynthesis operon. The P(3HB) content obtained with KEDA (pJC4) was lower than that obtained with its parent strain KS272 (pJC4). The reduced P(3HB) biosynthetic capacity of KEDA (pJC4) could be restored by the co-expression of the E. coli eda gene, which proves the important role of ED pathway on P(3HB) synthesis in recombinant E. coli as predicted by metabolic flux analysis. PMID- 12889026 TI - Comparisons of oxidative stress response genes in aerobic Escherichia coli fermentations. AB - The promoter regions of five SoxRS regulon genes (sodA, fumC, zwf, acnA, and acrAB) and one SoxRS regulatory protein gene (soxS) were inserted upstream of the gene of green fluorescent protein (GFP) in pGlow-TOPO. These promoter probe plasmids were transformed into Escherichia coli Top10 resulting in six strains that produce GFP in response to superoxide-induced stresses. Initial characterization from paraquat insults revealed significant induction of all six genes, with sodA, fumC, zwf, and soxS leading to the others in time and strength. These stress probe strains were then grown under similar conditions in fermentors and systematically exposed to varying durations of pure oxygen. Significant stimulation of the regulon was observed and quantitatively and temporally characterized by online monitoring of GFP fluorescence production (with transcriptional rate sodA > fumC > soxS, zwf > acnA, acrAB = 0). Interestingly, SoxRS regulon response occurred in typical E. coli fermentations where DO is maintained approximately 30% with increased agitation speed (with transcriptional rate acnA > sodA > zwf > acrAB > soxS, fumC = 0). These results also suggest that different molecular responses occur under different aeration schemes, all of which are intended to combat oxidative damage. PMID- 12889027 TI - Cometabolism of Cr(VI) by Shewanella oneidensis MR-1 produces cell-associated reduced chromium and inhibits growth. AB - Microbial reduction is a promising strategy for chromium remediation, but the effects of competing electron acceptors are still poorly understood. We investigated chromate (Cr(VI)) reduction in batch cultures of Shewanella oneidensis MR-1 under aerobic and denitrifying conditions and in the absence of an additional electron acceptor. Growth and Cr(VI) removal patterns suggested a cometabolic reduction; in the absence of nitrate or oxygen, MR-1 reduced Cr(VI), but without any increase in viable cell counts and rates gradually decreased when cells were respiked. Only a small fraction (1.6%) of the electrons from lactate were transferred to Cr(VI). The 48-h transformation capacity (Tc) was 0.78 mg (15 micromoles) Cr(VI) reduced. [mg protein](-1) for high levels of Cr(VI) added as a single spike. For low levels of Cr(VI) added sequentially, Tc increased to 3.33 mg (64 micromoles) Cr(VI) reduced. [mg protein](-1), indicating that it is limited by toxicity at higher concentrations. During denitrification and aerobic growth, MR-1 reduced Cr(VI), with much faster rates under denitrifying conditions. Cr(VI) had no effect on nitrate reduction at 6 microM, was strongly inhibitory at 45 microM, and stopped nitrate reduction above 200 microM. Cr(VI) had no effect on aerobic growth at 60 microM, but severely inhibited growth above 150 microM. A factor that likely plays a role in Cr(VI) toxicity is intracellular reduced chromium. Transmission electron microscopy (TEM) and electron energy loss spectroscopy (EELS) of denitrifying cells exposed to Cr(VI) showed reduced chromium precipitates both extracellularly on the cell surface and, for the first time, as electron-dense round globules inside cells. PMID- 12889028 TI - The effects of metabolites from the indigenous Acidithiobacillus thiooxidans and temperature on the bioleaching of cadmium from soil. AB - The effect of metabolites from the indigenous Acidithiobacillus thiooxidans and temperature on the bioleaching of cadmium from soil was investigated in the present study. Bioleaching was found to be more effective than chemical leaching of cadmium. The metabolite, mainly sulfuric acid, which was shown to be growth associated in the exponential phase, plays a major role in bioleaching. The maximum amount of cadmium leached was obtained after 8 days of precultivation when cells were directly involved in the leaching process. It indicates that cells in the exponential growth phase exhibit higher activity toward bioleaching. In contrast, the maximum amount of cadmium leached and the maximum initial rate for bioleaching were reached after 16 days of precultivation when only metabolites were involved in the bioleaching process. It implies that higher sulfuric acid concentration results in higher leaching efficiency. In addition, higher temperature leads to higher leaching efficiency. The optimal operation condition for bioleaching was determined to be a two-stage process: The first stage involves the precultivation of the indigenous A. thiooxidans at 30 degrees C for 8 days followed by 20 minutes of centrifugation to discard cells. The second stage involves the bioleaching with the subsequent supernatant at 50 degrees C. PMID- 12889029 TI - Chitosan scaffolds for biomolecular assembly: coupling nucleic acid probes for detecting hybridization. AB - Chitosan, a naturally occurring biopolymer, was used as a scaffold for the covalent binding of single-stranded DNA oligonucleotide probes in a fluorescence based nucleic acid hybridization assay. Chitosan's pH dependent chemical and electrostatic properties enable its deposition on electrodes and metal surfaces, as well as on the bottom of microtiter plates. A combinatorial 96-well microtiter plate format was used to optimize chemistries and reaction conditions leading to hybridization experiments. We found the coupling of oligonucleotides using relatively common glutaraldehyde chemistry was quite robust. Our hybridization results for complementary ssDNA oligonucleotides (E. coli dnaK sequences) demonstrated linear fluorescence intensity with concentration of E. coli dnaK specific oligonucleotide from 0.73 microM to 6.6 microM. Moreover, hybridization assays were specific as there was minimal fluorescence associated with noncomplementary groEL oligonucleotide. Finally, these results demonstrate the portability of a DNA hybridization assay based on covalent coupling to chitosan, which, in turn, can be deposited onto various surfaces. More arduous surface preparation techniques involving silanizing agents and hazardous washing reagents are eliminated using this technique. PMID- 12889030 TI - Scale-up of Artemisia annua L. hairy root cultures produces complex patterns of terpenoid gene expression. AB - Hairy roots grow quickly, reach high densities, and can produce significant amounts of secondary metabolites, yet their scale-up to bioreactors remains challenging. Artemisia annua produces a rich array of terpenoids, including the sesquiterpene, artemisinin, and transformed roots of this species provide a good model for studying terpenoid production. These cultures were examined in shake flasks and compared with cultures grown in two types of bioreactors, a mist reactor and a bubble column reactor, which provide very different environments for the growing roots. Mist reactors have been shown previously to result in cultures that produce significantly more artemisinin per gram fresh weight of culture, while bubble column reactors have produced greater biomass. We have compared expression levels of four key terpenoid biosynthetic genes: 3-hydroxy-3 methylglutaryl coenzyme A reductase (HMGR), 1-deoxy-D-xylulose-5-phosphate synthase (DXS), 1-deoxy-D-xylulose-5-phosphate reductoisomerase (DXR), and farnesyl diphosphate synthase (FPS) in the three culture conditions. In shake flasks we found that although all four genes showed temporal regulation, only FPS expression correlated with artemisinin production. Light also affected the transcription of all four genes. Although expression in reactors was equivalent to or greater than that of roots grown in shake flasks, no correlation was found between expression level within six different zones of each reactor and their respective oxygen levels, light, and root-packing density. Surprisingly, transcriptional regulation of HMGR, DXS, DXR, and FPS was greatly affected by the position of the roots in each reactor. Thus, relying on a single reactor sample to characterize the gene activity in a whole reactor can be misleading, especially if the goal is to examine the difference between reactor types or operating parameters, steps essential in scaling up cultures for production. PMID- 12889031 TI - On-line monitoring of Phaffia rhodozyma fed-batch process with in situ dispersive Raman spectroscopy. AB - Since the yeast Phaffia rhodozyma was first described some 35 years ago, there has been significant interest in the development of commercial processes to exploit its ability to produce carotenoids (approximately 80% astaxanthin). However, the optimal conditions for carotenoid production are not well understood. A key limitation has been the lack of an appropriate sensor for on line carotenoid quantification. In this study, an in situ Raman spectroscopy probe was used to monitor intracellular carotenoid production for three consecutive P. rhodozyma fed-batch experiments. Raman spectroscopy is particularly well suited to the study of carotenoids due to a resonance effect, which greatly enhances the intensity of the three fundamental carotenoid bands, nu(1) (1513 cm(-1), C(-) (-)C stretch), nu(2) (1154 cm(-1), C-C stretch), and nu(3) (1003 cm(-1), CH(3) rock). For all three cultures, the peak height of these bands was linearly correlated with intracellular carotenoid content (1 to 45 mg/L) to a precision of better than 5%, and the correlation from one experiment was directly applicable to others. PMID- 12889032 TI - Scalable inoculation strategies for microcarrier-based animal cell bioprocesses. AB - Scalability is a major demand for high-yield, stable bioprocess systems in animal cell culture-based biopharmaceutical production. Increased yields can be achieved through high-density cell culture, such as in the combination of microcarrier and fluidized bed bioreactor technology. To minimize inocula volume in industrial applications of fluidized bed fermentation systems, it is crucial to increase the bed volume in the reactor during the fermentation process. We tested scale-up strategy for the production of recombinant human arylsulfatase B (ASB) enzyme used in enzyme replacement therapy in patients afflicted with mucopolysaccharidosis type VI (MPS VI). This enzyme was derived from Chinese hamster ovary (CHO) cells cultivated as adherent cell culture on Cytoline macroporous microcarriers (Amersham Biosciences, Uppsala, Sweden) using a Cytopilot Mini fluidized bed bioreactor (FBR; Amersham Biosciences, Vogelbusch, Austria). Both 1:2 expansion (herein referred to as the addition of fresh, not yet-colonized microcarriers) and 1:6 expansion of the carrier bed were performed successfully; the cells restarted to proliferate for colonizing these newly added carriers; and the stability of the culture was not negatively affected. PMID- 12889033 TI - Expression of galP and glk in a Escherichia coli PTS mutant restores glucose transport and increases glycolytic flux to fermentation products. AB - In Escherichia coli, the uptake and phosphorylation of glucose is carried out mainly by the phosphotransferase system (PTS). Despite the efficiency of glucose transport by PTS, the required consumption of 1 mol of phosphoenolpyruvate (PEP) for each mol of internalized glucose represents a drawback for some biotechnological applications where PEP is a precursor of the desired product. For this reason, there is considerable interest in the generation of strains that can transport glucose efficiently by a non-PTS mechanism. The purpose of this work was to study the effect of different gene expression levels, of galactose permease (GalP) and glucokinase (Glk), on glucose internalization and phosphorylation in a E. coli PTS(-) strain. The W3110 PTS(-), designated VH32, showed limited growth on glucose with a specific growth rate (mu) of 0.03 h(-1). A low copy plasmid family was constructed containing E. coli galP and glk genes, individually or combined, under the control of a trc-derived promoter set. This plasmid family was used to transform the VH32 strain, each plasmid having different levels of expression of galP and glk. Experiments in minimal medium with glucose showed that expression of only galP under the control of a wild-type trc promoter resulted in a mu of 0.55 h(-1), corresponding to 89% of the mu measured for W3110 (0.62 h(-1)). In contrast, no increase in specific growth rate (mu) was observed in VH32 with a plasmid expressing only glk from the same promoter. Strains transformed with part of the plasmid family, containing both galP and glk genes, showed a mu value similar to that of W3110. Fermentor experiments with the VH32 strain harboring plasmids pv1Glk1GalP, pv4Glk5GalP, and pv5Glk5GalP showed that specific acetate productivity was twofold higher than in W3110. Introduction of plasmid pLOI1594, coding for pyruvate decarboxylase and alcohol dehydrogenase from Zymomonas mobilis, to strain VH32 carrying one of the plasmids with galP and glk caused a twofold increase in ethanol productivity over strain W3110, also containing pLOI1594. PMID- 12889034 TI - Effect of silkworm hemolymph on N-linked glycosylation in two Trichoplusia ni insect cell lines. AB - A recombinant N-linked glycoprotein, secreted human placental alkaline phosphatase (SEAP), was produced in two Trichoplusia ni insect cell lines using the baculovirus expression vector. Silkworm hemolymph (SH) was added to TNMFH + 10% fetal bovine serum (FBS) medium to a concentration of 2.5% or 5%, and SEAP production and glycosylation in the presence of SH were compared with controls devoid of hemolymph. Growing Tn-4s cells in 5% SH-supplemented medium required progressive adaptation of the cells to SH, and adapted cells had a SEAP specific yield decreased by 2.5-fold compared with control cells not exposed to SH. Although SEAP produced in the control possessed little complex glycosylation (<1%), SEAP produced by SH-adapted cells in the presence of 5% SH possessed 8.7% sialylated structures, as well as unusual, asialylated, agalactosylated structures with a high degree of polymerization (DP). On the basis of enzymatic and mass-spectrometric analyses, we propose that these structures are glucosylated, high-mannose oligosaccharides. SEAP was also produced by Tn-4s cells without adaptation to SH when SH was added just prior to baculovirus infection, but SEAP specific yield was adversely affected (approximately fourfold reduction compared with control devoid of hemolymph), and glycosylation of SEAP produced under these conditions was characterized by large amounts of high mannose and high-DP structures and an absence of complex structures. Similarly, Tn5B1-4 cells that were not adapted to SH had a SEAP specific yield reduced by approximately fivefold in SH-containing medium; however, these cells were able to produce 13.5% sialylated SEAP in the presence of 2.5% SH, whereas complex structures were not produced in the absence of SH. We propose that SH improves glycosylation either directly or indirectly by decreasing SEAP specific yield. PMID- 12889035 TI - Simultaneous nitrification and denitrification using stored substrate (PHB) as the electron donor in an SBR. AB - The potential for PHB (poly-beta-hydroxybutyrate) to serve as the electron donor for effective simultaneous nitrification and denitrification (SND) was investigated in a 2-L sequencing batch reactor (SBR) using a mixed culture and acetate as the organic substrate. During the feast period (i.e., acetate present), heterotrophic respiration activity was high and nitrification was prevented due to the inability of nitrifying bacteria to compete with heterotrophs for oxygen. Once acetate was depleted the oxidation rate of PHB was up to 6 times slower than that of soluble acetate and nitrification could proceed due to the decreased competition for oxygen. The slow nature of PHB degradation meant that it was an effective substrate for SND, as it was oxidised at a similar rate to ammonium and was therefore available for SND throughout the entire aerobic period. The percentage of nitrogen removed via SND increased at lower DO concentrations during the famine period, with up to 78% SND achieved at a DO concentration of 0.5 mg L(-1). However, the increased percentage of SND at a low DO concentration was compromised by a 2-times slower rate of nitrogen removal. A moderate DO concentration of 1 mg L(-1) was optimal for both SND efficiency (61%) and rate (4.4 mmol N x Cmol x(-1) x h(-1)). Electron flux analysis showed that the period of highest SND activity occurred during the first hour of the aerobic famine period, when the specific oxygen uptake rate (SOUR) was highest. It is postulated that a high SOUR due to NH(4) (+) and PHB oxidation decreases oxygen penetration into the floc, creating larger zones for anoxic denitrification. The accumulation of nitrate towards the end of the SND period showed that SND was finally limited by the rate of denitrification. As PHB degradation was found to follow first-order kinetics (df(PHB)/dt = -0.19 x f(PHB)), higher PHB concentrations would be expected to drive SND faster by increasing the availability rate of reducing power and reducing penetration of oxygen into the floc, due to the corresponding increased SOUR. Process control techniques to accumulate higher internal PHB concentrations to improve PHB-driven SND are discussed. PMID- 12889036 TI - Detection and identification of Escherichia coli, Shigella, and Salmonella by microarrays using the gyrB gene. AB - Commonly, 16S ribosome RNA (16S rRNA) sequence analysis has been used for identifying enteric bacteria. However, it may not always be applicable for distinguishing closely related bacteria. Therefore, we selected gyrB genes that encode the subunit B protein of DNA gyrase (a topoisomerase type II protein) as target genes. The molecular evolution rate of gyrB genes is higher than that of 16S rRNA, and gyrB genes are distributed universally among bacterial species. Microarray technology includes the methods of arraying cDNA or oligonucleotides on substrates such as glass slides while acquiring a lot of information simultaneously. Thus, it is possible to identify the enteric bacteria easily using microarray technology. We devised a simple method of rapidly identifying bacterial species through the combined use of gyrB genes and microarrays. Closely related bacteria were not identified at the species level using 16S rRNA sequence analysis, whereas they were identified at the species level based on the reaction patterns of oligonucleotides on our microarrays using gyrB genes. PMID- 12889037 TI - Global incorporation of norleucine in place of methionine in cytochrome P450 BM-3 heme domain increases peroxygenase activity. AB - In this study we have replaced all 13 methionine residues in the cytochrome P450 BM-3 heme domain (463 amino acids) with the isosteric methionine analog norleucine. This experiment has provided a means of testing the functional limits of globally incorporating into an enzyme an unnatural amino acid in place of its natural analog, and also an efficient way to test whether inactivation during peroxide-driven P450 catalysis involves methionine oxidation. Although there was no increase in the stability of the P450 under standard reaction conditions (in 10 mM hydrogen peroxide), complete substitution with norleucine resulted in nearly two-fold-increased peroxygenase activity. Thermostability was significantly reduced. The fact that the enzyme can tolerate such extensive amino acid replacement suggests that we can engineer enzymes with unique chemical properties via incorporation of unnatural amino acids while retaining or improving catalytic properties. This system also provides a platform for directing enzyme evolution using an extended set of protein building blocks. PMID- 12889038 TI - A novel method of simulating oxygen mass transfer in two-phase partitioning bioreactors. AB - An empirical correlation, based on conventional forms, has been developed to represent the oxygen mass transfer coefficient as a function of operating conditions and organic fraction in two-phase, aqueous-organic dispersions. Such dispersions are characteristic of two-phase partitioning bioreactors, which have found increasing application for the biodegradation of toxic substrates. In this work, a critical distinction is made between the oxygen mass transfer coefficient, k(L)a, and the oxygen mass transfer rate. With an increasing organic fraction, the mass transfer coefficient decreases, whereas the oxygen transfer rate is predicted to increase to an optimal value. Use of the correlation assumes that the two-phase dispersion behaves as a single homogeneous phase with physical properties equivalent to the weighted volume-averaged values of the phases. The addition of a second, immiscible liquid phase with a high solubility of oxygen to an aqueous medium increases the oxygen solubility of the system. It is the increase in oxygen solubility that provides the potential for oxygen mass transfer rate enhancement. For the case studied in which n-hexadecane is selected as the second liquid phase, additions of up to 33% organic volume lead to significant increases in oxygen mass transfer rate, with an optimal increase of 58.5% predicted using a 27% organic phase volume. For this system, the predicted oxygen mass transfer enhancements due to organic-phase addition are found to be insensitive to the other operating variables, suggesting that organic-phase addition is always a viable option for oxygen mass transfer rate enhancement. PMID- 12889039 TI - Effects of beta-cyclodextrin-dextran polymer on stability properties of trypsin. AB - Dextran modified with the mono-6-pentylene-diamino-6-deoxy-beta-cyclodextrin derivative was evaluated as a thermoprotectant additive for trypsin. The optimum temperature for trypsin activity was increased by 7 degrees C in the presence of this polymer. The enzyme thermostability was increased from 48.5 to 64 degrees C over 10 min of incubation, and the activation free energy of thermoinactivation at 50 degrees C was increased by 4.1 kJ/mol in the presence of the additive. Trypsin was 6-fold more resistant to autolytic inactivation at alkaline pH in the presence of the polymer. PMID- 12889040 TI - Epithelioid variant of gastrointestinal stromal tumor: Diagnosis by fine-needle aspiration. AB - Epithelioid gastrointestinal stromal tumors (GISTs) may cause significant diagnostic confusion on fine-needle aspiration (FNA) with carcinomas, neuroendocrine tumors, and melanoma, particularly when metastatic. This study characterizes the cytologic features of nine cases of epithelioid GISTs that were obtained by computerized tomographic guidance in five, by endoscopic ultrasound in three, and from an excised liver tumor in one. Six cases presented as liver masses, one as a perisplenic mass, one as an abdominal mass, and one as a gastric mass. The aspirates revealed mainly single or small clusters of epithelioid cells with a moderate amount of granular to clear cytoplasm, small uniform nuclei with mild to marked nuclear envelope irregularities. Binucleation and intranuclear inclusions were frequent findings. Collagenous stroma was seen in most cases. In three cases, a neuroendocrine tumor was the initial diagnosis. Immunocytochemical staining for c-kit (CD117) was performed on cellblocks in six cases and was positive in five cases. On the subsequent surgical specimen, CD117 was positive in the c-kit-negative cytology case. The diagnosis of GIST should be considered in aspirates of the gastrointestinal tract, liver, mesentery, or abdominal wall mass lesions when epithelioid cells are the predominant cell type. Ancillary studies such as immunohistochemical stains are usually helpful in making a definitive diagnosis. PMID- 12889041 TI - Detection of a subset of CD30+ anaplastic large cell lymphoma by interphase fluorescence in situ hybridization. AB - T/null-cell anaplastic large cell lymphoma (ALCL) is a morphologically and clinically heterogeneous group of non-Hodgkin's lymphoma; to date several morphologic variants have been described on histologic specimens. However, the cytologic features of these variants in the fine-needle aspiration (FNA) specimens have not been well evaluated. The t(2;5)(p23;q35) has been identified in a subset of T/null-ALCL and is known to be associated with a favorable prognosis. We reviewed the cytomorphologic characteristics in 24 FNA specimens of ALCL. In all cases, the diagnosis was confirmed on histologic specimens, and immunohistochemical studies for anaplastic lymphoma kinase (ALK) protein expression were performed on the aspirates. The presence of ALK breakpoints were evaluated in nine cases, using a DNA break-apart probe on chromosome 2 covering the ALK gene by fluorescence in situ hybridization (FISH) techniques. Two hundred cells per case were examined. The results were expressed as the percentage of cells containing more than two signals of chromosome 2 to the total number of cells counted. FNA sites included lymph nodes (20), lung (2), breast (1), and soft tissue (1). The median age of the patients was 56 yr (range, 17-75 yr). Twenty cases had systemic involvement; in four cases, skin was the primary site with secondary involvement of the lymph nodes. All cases were CD30(+) by immunohistochemistry; 20 were of T-cell phenotype and 4 were null cell type. The cytologic evaluation revealed typical anaplastic morphology (common type) with many "hallmark cells" in 16 (67%) cases. Other morphologic variants identified were small cell pattern in five cases, monomorphic pattern in two cases, and lymphohistiocytic pattern in one case. FISH studies showed that six (66.7%) of nine cases had at least two signals of chromosome 2, consistent with ALK breakpoints. With careful cytomorphologic evaluation in conjunction with appropriate immunohistochemical studies, a diagnosis of ALCL can be confidently made in the FNA specimens in the cellular aspirates and its morphologic variants also can be recognized. Furthermore, the FNA specimen is suitable in detecting ALK breakpoints by FISH study, permitting rapid identification of a subset of patients with ALCL, who may have a favorable prognosis. Using a commercially available probe, detection of ALK breakpoints in the FNA specimens is simple and can be a useful diagnostic adjunct in cases where distinction from other lymphomas or lymphoid lesions is morphologically difficult. PMID- 12889042 TI - Urine cytology in renal glomerular disease and value of G1 cell in the diagnosis of glomerular bleeding. AB - The objectives of the present study were to evaluate the cytology of urine sediments in patients with glomerular diseases, as well as the value of G1 dysmorphic erythrocytes (G1DE) or G1 cells in the detection of renal glomerular hematuria. Freshly voided urine samples from 174 patients with glomerular diseases were processed according to the method used for semiquantitative cytologic urinalysis. G1DEs (distorted erythrocytes with doughnut-like shape, target configuration with or without membranous protrusions or blebs), non-G1DEs (distorted erythrocytes without the above-mentioned morphologic changes), normal erythrocytes (NEs), and renal tubular cells (RTCs) were evaluated. Erythrocytic casts (ECs) were counted and graded as abundant (>1 per high-power field) or rare (1 per 5 high-power fields). G1DE/total erythrocyte ratios were calculated by counting 200 erythrocytes including G1DEs, non-G1DEs, and NEs. Only abundant NEs were found in 13 cases; abundant G1DEs, non-G1DEs, NEs, and no ECs in 95 cases; abundant NEs, non-G1DEs, and ECs and no G1DEs in 31 cases; and abundant NEs, G1DEs and non-G1DEs, and rare ECs in 35 cases. In 130 cases in which G1DEs were present, the G1DE/total erythrocyte ratios varied from 10% to 100%. This parameter was greater or equal to 80%, 50%, 20%, and 10% in 58 (44.6%), 29 (22.3%), 28 (21.5%), and 15 (11.5%) patients, respectively. In all cases, the number of RTCs was within normal limits or slightly increased, and a variable number of non-G1DEs were present in 161 cases. Thus, abundant ECs and/or G1DEs with a G1DE/total erythrocyte ratio of 10-100% proved to be specific urinary markers for renal glomerular diseases. PMID- 12889043 TI - Flow cytometry as an adjunct to cytomorphologic analysis of serous effusions. AB - The role of flow cytometry (FC) in the diagnosis of lymphoid lesions by fine needle aspiration (FNA) is well established. However, studies evaluating the usefulness of FC in serous cavity effusions (SCE) are few. We performed a retrospective review of 115 consecutive SCE with concurrent FC analysis, comparing the provisional cytopathologic diagnosis (PCD), i.e., before the FC results were added, with final diagnoses as modified by subsequent FC immunophenotyping. The predominant clinical indication for the FC analysis was the presence of a spontaneous SCE in a patient with a history of malignant lymphoma. Three- or four-color analysis was performed using antibodies against CD45, CD71, CD33, CD22, CD19, CD20, kappa, lambda, CD5, CD3, and CD56. The PCD was benign in 47%, atypical in 16%, and malignant in 37% of cases. The latter category consisted mostly of malignant lymphoma (n = 32), but also included acute lymphoblastic leukemia (1 case), T-cell lymphoma/leukemia (2 cases), acute myelogenous leukemia (1 case), multiple myeloma (1 case), Hodgkin's lymphoma (1 case), sarcoma (1 case), and adenocarcinoma (4 cases). In 18 cases (16%), the PCD was later modified by the FC results from atypical/suspicious to benign (8) and from benign or atypical/suspicious to malignant (10 cases). The latter group included acute natural killer (NK) cell leukemia (1 case), chronic lymphocytic leukemia (1 case), mantle cell lymphoma (2 cases), follicular lymphoma (3 cases), angioimmunoblastic lymphoma (1 case), large cell lymphoma (1 case), and multiple myeloma (1 case). As expected, FC was noncontributory in cases of Hodgkin's lymphoma and nonlymphoid malignancies. In summary, immunophenotyping by FC modified the PCD significantly in 16% of SCE, permitting appropriate cancer staging and management. The above data underscore the importance of FC as an adjunct to cytomorphology in SCE. PMID- 12889044 TI - Accuracy of cytological grading of spindle-cell sarcomas. AB - Spindle-cell sarcomas constitute an important component of soft-tissue sarcomas, where accurate grading may be more important than histologic subtype in order to plan treatment strategies and determine prognosis. To evaluate the applicability and accuracy of grading spindle-cell sarcomas on fine-needle aspiration cytology (FNAC) smears, 54 cases of histologically documented spindle-cell sarcomas, consisting of synovial sarcomas (20 cases), neurofibrosarcomas (12 cases), leiomyosarcomas (9 cases), dermatofibrosarcoma protuberans (DFSP; 6 cases), fibrosarcomas (3 cases), hemangiopericytomas (2 cases), and spindle-cell sarcomas, unclassified (2 cases), were graded according to a three-tier system proposed earlier for FNAC smears, while the histological sections were graded using the French Federation of National Cancer Centers (FNCLCC) grading system. The cytological grading was correlated with the histological grade. There was an overall cytologic and histologic concordance in 40/54 (74%) cases, and concordance in 9/13 (69%) grade I, 19/25 (76%) grade II, and 12/16 (75%) grade III cases. Analysis of grading of individual sarcomas revealed a concordance in 92% of neurofibrosarcomas, 78% of leiomyosarcomas, 70% of synovial sarcomas, 67% of dermatofibrosarcoma protuberans, 67% of fibrosarcomas, 50% of hemangiopericytomas, and 50% of cases of malignant mesenchymal tumors, spindle cell type unclassified. Major noncorrelation was seen in 10/54 (18.5%) cases. Minor noncorrelation was seen in 4/54 (7.4%) cases. Thus it is possible to accurately predict the grade in 74% of cases of spindle-cell sarcomas. The cytological and histological concordance was better (75%) in high-grade (grades II and III) as compared to grade I sarcomas (69%). Sampling errors due to morphologic heterogeneity in sarcomas may cause noncorrelation in a few cases. PMID- 12889045 TI - Cytologic diagnosis of molluscum contagiosum in scrape samples from facial lesions. PMID- 12889046 TI - Fractal dimensions of breast lesions on cytology smears. AB - In the present study, box-counting fractal dimensions of benign and malignant cells of breast tumors on cytology material were measured and compared. We selected fine-needle aspiration cytology smears of 14 cases of histopathology proven infiltrating duct carcinomas and 7 cases of fibroadenoma of the breast. Five cells were randomly selected from each case. Box-counting fractal dimensions of all cells were measured with the help of an image cytometer (Leica, Cambridge, UK), using Quantimet 600 software (Leica). In total, 70 malignant cells and 35 benign cells were studied. The mean fractal dimensions of malignant cells and benign cells were 0.9571 +/- 0.1265 and 0.8354 +/- 0.1367, respectively. There was significant difference in the fractal dimension of malignant and benign cells (P = 0.006, Mann-Whitney nonparametric test). The measurement of fractal dimension may be helpful in discriminating malignant from benign cells. This may be another discriminating feature of malignant cells, along with classic image morphometry based on Euclidean geometry. PMID- 12889047 TI - Case of adrenal lymphangioma with atypical lymphocytes in aspirate cytology. AB - Cytological diagnosis of adrenal cysts is becoming more important, with the increase in cases identified due to the widespread use of imaging modalities. Adrenal cysts have been classified into four main groups: endothelial, epithelial, parasitic, and pseudocyst, with endothelial cysts further classified as lymphangiomatous or angiomatous. We report on a case of adrenal lymphangioma with atypical lymphocytes found in the intraoperative aspirate, suggesting an inconclusive diagnosis, although immunohistochemical study led us to suppose their B-cell origin. This case provided important cytological findings that may assist in the prevention of cytological overdiagnosis. PMID- 12889048 TI - Metastatic islet cell carcinoma to the umbilicus: diagnosis by fine-needle aspiration. AB - Internal malignancies rarely metastasize to the umbilical region. The gastrointestinal tract and female genital tract are the most common primary sites. Although the pancreas is frequently involved, the majority of these metastases are adenocarcinoma. Here, we present a case of umbilical metastasis from an islet cell carcinoma diagnosed by fine-needle aspiration in a young woman 2 yr after distal pancreatectomy. This finding led to further radiological studies and surgical intervention, which revealed intraabdominal tumor spread. Neuroendocrine tumors represent a rare tumor type that can give rise to umbilical metastases. Only two cases of carcinoid tumor metastasizing to the umbilicus have been reported. PMID- 12889049 TI - Cellular schwannoma: report of a case diagnosed intraoperatively with the aid of cytologic imprints. AB - The diagnosis of cellular schwannoma in intraoperative consultation is difficult because of the hypercellularity and deep location, which may lead to errors. The recognition of cytologic features together with the histological appearance are of great importance to make the correct diagnosis. An accurate diagnosis of this variant of schwannoma is important because other spindle cell tumors display differences in treatment and clinical behavior. We report a demonstrative case of cellular schwannoma arising in retroperitoneum of a 52-yr-old woman in which the cytologic imprints were useful to make an accurate diagnosis intraoperatively. Briefly, the differential diagnosis is discussed. PMID- 12889050 TI - Aspiration cytology of ameloblastic fibroma: a diagnostic challenge. AB - Ameloblastic fibroma of the jaw is a rare, benign mixed odontogenic tumor, having little tendency for local invasion and a low recurrence rate. Cytologic distinction from ameloblastoma, ameloblastic fibrosarcoma, and intraosseous adenoid cystic carcinoma is necessary, in view of the different biologic behavior. A painful, slow-growing swelling of the jaw in a 5-yr-old child clinicoradiologically considered as a benign cystic lesion was aspirated. Sheets of small monomorphic epithelial cells with peripheral palisading by columnar cells were seen on cytology smears. The striking feature was central hyaline globules in some tubules. A cytologic possibility of adenomatoid odontogenic tumor was suggested. Histopathology, however, confirmed it to be an ameloblastic fibroma. PMID- 12889051 TI - Fine-needle aspiration cytology of giant cell tumor of tendon sheath. AB - Giant cell tumor of tendon sheath (GCTTS) is a unique soft tissue lesion of the hands and feet. As the cytomorphological features of this lesion are rarely documented, the spectrum of cytomorphological features in 20 cases of GCTTS seen in fine-needle aspiration (FNA) smears are presented. Patients were in the 12-64 yr age group with an equal sex ratio. Fingers or thumb were the commonest site (16 cases), followed by foot (3 cases) and palm (1 case). FNA smears were cellular and composed of varying proportions of stromal and giant cells. Stromal cells showed a dispersed arrangement and were polygonal to spindle shaped. Nuclear grooves and convolutions were found in some of the stromal cells in all cases. Intranuclear cytoplasmic inclusions were occasionally seen. Polygonal cells with round nuclei and nucleoli having abundant cytoplasm, along with binucleate forms, were also found in all cases. Histological sections were available in 10 cases and corroborated the cytological features. Hemosiderin laden macrophages (11 of 20 cases) and abundant foamy vacuolation of stromal cells (3 of 20 cases) were also observed in FNA smears. In tissue sections, both features were seen in all cases, but with a patchy distribution. The cytological features of GCTTS are uniform, and FNA cytodiagnosis is possible. PMID- 12889052 TI - Pap test discrepancies and follow-up histology. AB - Papanicolaou (Pap) test discrepancy rates between cytotechnologists (CTs) and cytopathologists (CPs) are often kept to evaluate the performance of individual CTs. This is based on the unproven assumption that the CP's diagnoses are more likely to be correct. We investigated this assumption using data from our discrepancy files and comparing them to follow-up histology. All Pap test discrepancies were noted between January 1, 2001-December 31, 2001. Surgical pathology files were then searched for follow-up histology within 9 mo of the Pap test. Histologic diagnoses were compared with the previous CT and CP diagnoses, and then judged regarding accuracy. In total, 63,376 Pap tests were evaluated between January 1, 2001-December 31, 2001. There were 795 discrepancies throughout this period (1.25%). One hundred and sixty-six cases with discrepancies had follow-up histology within 9 mo of the Pap test (20.9%). Of downgraded cases (103), CPs were more correct in 51 cases (49.5%), whereas CTs were more correct in 52 cases (50.5%). Of upgraded cases (63), CPs were more correct in 19 cases (30.2%), whereas CTs were more correct in 44 cases (69.8%). Our results suggest that CPs are not more likely to be correct than CTs when there is a discrepancy with the diagnosis of a Pap test, especially when CPs upgrade CT diagnoses. This suggests that discrepancy data may be helpful for evaluating the performance of both CPs and CTs. It may also be of educational use for both CPs and CTs to know the follow-up histology in these cases. PMID- 12889053 TI - Cytologic diagnosis of hemangiopericytoma of nasal cavity. PMID- 12889054 TI - Pleural mesothelioma presenting initially as empyema. PMID- 12889056 TI - Morphogenesis of blood vessels in the head muscles of avian embryo: spatial, temporal, and VEGF expression analyses. AB - Adult skeletal muscle is a highly vascularized tissue, but the development of intramuscular endothelial networks has not been well studied. In quail embryos, QH1-positive angioblasts are present and moving throughout myogenic head mesoderm before the onset of primary myotube formation. On day 5 of incubation, concurrent with early myotube formation and aggregation, angioblasts establish a transient vascular plexus surrounding the myogenic condensations. Between days 5 and 9, the intramuscular vessels form an irregular network of endothelial cords and patent channels and only later are the parallel arrays of capillaries characteristic of adult muscles established. Microinjections using India ink, QH1, and Mercox resin reveal that these intramuscular capillaries are typically not connected to systemic vessels of the head until day 10, which is near the end of primary myogenesis and corresponds to the onset of muscular function. Morphometric analyses performed during primary myogenesis stages show a decrease in muscle cell density but no significant changes in intramuscular vascular density between days 5 and 9. This finding was surprising, as it is generally assumed that muscle growth requires elevated oxygen and nutrient levels. Moreover, there are no significant morphometric differences in vascular supply to embryonic fast and slow muscles. Endothelial tissue density is similar in slow muscles (oculorotatory, e.g., lateral rectus), fast muscles (mandibular depressor), and mixed muscles, in which the fiber types can be interspersed (jaw adductors) or segregated (branchiomandibular). Vascular endothelial growth factor (VEGF) protein is abundant in myotubes but not endothelial cells within both fast and slow head muscles at days 7 and 9. However, in some mixed muscles, only a minority of myotubes, which do not correspond to one specific fiber type, express VEGF. These results document a dynamic set of intramuscular and perimuscular angiogenic reorganizations during avian head myogenesis. Thus far, no vasculogenic distinctions between fast and slow muscles have been observed, although muscle heterogeneity in VEGF expression is evident. PMID- 12889057 TI - In vivo regulation of the chicken cardiac troponin T gene promoter in zebrafish embryos. AB - The chicken cardiac troponin T (cTnT) gene is representative of numerous cardiac and skeletal muscle-specific genes that contain muscle-CAT (MCAT) elements within their promoters. We examined the regulation of the chicken cTnT gene in vivo in zebrafish embryos, and in vitro in cardiomyocyte, myoblast, and fibroblast cultures. Defined regions of the cTnT promoter were linked to the green fluorescent protein (GFP) gene for in vivo analysis, and the luciferase gene for in vitro analysis. Injection of the cTnT promoter constructs into fertilized zebrafish eggs resulted in GFP expression in both heart and skeletal muscle cells reproducing the pattern of expression of the endogenous cTnT gene in the chicken embryo. Promoter deletion analysis revealed that the cis-regulatory regions responsible for cardiac and skeletal muscle-specific expression functioned in an equivalent manner in both in vitro and in vivo environments. In addition, we show that mutation of the poly-ADP ribose polymerase-I (PARP-I) binding site adjacent to the distal MCAT element in the chicken cTnT promoter produced a non-cell specific promoter in vitro and in the zebrafish. Thus, the PARP-I transcriptional regulatory mechanism that governs muscle specificity of the chicken cTnT promoter is conserved across several chordate classes spanning at least 350 million years of evolution. PMID- 12889058 TI - Pigment cell organization in the hypodermis of zebrafish. AB - Zebrafish have a characteristic horizontal-stripe pigment pattern made by a specific distribution of three types of pigment cells: melanophores, xanthophores, and iridophores. This pattern is a valuable model to investigate how the spatial patterns form during animal development. Although recent findings suggest that the interactions among the pigment cells play a key role, the particular details of these interactions have not yet been clarified. In this report, we performed transmission electron microscopic study to show the distribution, conformation, and how the cells contact with each other in the hypodermis. We found that the pigment cells form complex but ordered, layered structures in both stripe and interstripe regions. The order of the layered structures is kept strictly all through the hypodermal regions. Our study will provide basic information to investigate the mechanism of pigment pattern formation in zebrafish. PMID- 12889059 TI - Pluripotency of reprogrammed somatic genomes in embryonic stem hybrid cells. AB - Somatic nuclei can be epigenetically reprogrammed by factors present in undifferentiated embryonic stem (ES) cells. The acquisition of pluripotency by somatic genomes could render such cells a viable source of personalized cell type(s) for therapeutic application, avoiding the need for controversial therapeutic cloning. To investigate this possibility, we first determined the origin of transcripts in teratomas generated from mouse (ES x somatic cell) hybrid clones. Transcription of markers from the somatic genome demonstrated efficient in vivo differentiation down independent lineages. The induction of dopaminergic neurons by coculture with stromal PA6 feeder cells also demonstrated efficient capacity to differentiate in vitro. Hybrid clone-derived neurons expressed appropriate markers, and transcription of Pitx3 from the somatic genome was confirmed. When transplanted into mouse brains, the dopaminergic neurons were successfully integrated and expressed tyrosine hydroxylase. Thus, it should be possible to produce personalized ES-like cells with the reprogrammed somatic genomes. PMID- 12889060 TI - Evidence for an extracellular matrix bridge guiding proepicardial cell migration to the myocardium of chick embryos. AB - During heart development, the proepicardium (PE) gives rise to cells of the epicardial epithelium, connective tissue of the subepicardium and the myocardium, and smooth muscle, endothelium, and connective tissue of the coronary arteries. The PE arises as an outgrowth of the pericardial serosa at embryonic day 2 (Hamburger and Hamilton stage [HH] 14) of chick development. Between stages HH14 and HH17, multicellular villous projections extend from the PE toward the dorsal aspect of the lesser curvature of the myocardium. On reaching the atrioventricular (AV) junction, the cells spread over the myocardium, eventually enveloping the complete heart surface as a simple squamous epithelium. Although the lineage of the PE cells is well established, it remains uncertain how cells of the PE reach the myocardial surface and specifically target the AV junction. By using a combination of serial section reconstructions, immunofluorescence, and electron microscopy, we have identified an extracellular matrix bridge (ECMB) spanning the coelomic cavity between the PE and the myocardium. The ECMB is first detectable at HH14 and persists until the PE contacts the bare myocardial surface. This ECMB stains intensely with ruthenium red and Alcian blue, contains heparan sulfate and fibronectin, and exhibits both fibrillar and globular ultrastructure, reminiscent of proteoglycans. After PE attachment to the myocardium (HH16-HH17), the subepicardium exhibited strong staining for heparan sulfate. Heparinase injection into the pericardial coelom at HH15 resulted in aberrant development of the primordial epicardium. On the basis of these studies, we suggest that the ECMB may participate in migration and targeting of the PE to the myocardium. PMID- 12889061 TI - Selective control of neuronal cluster size at the forebrain/midbrain boundary by signaling from the prechordal plate. AB - Within the vertebrate embryonic neural plate, the first neuronal clusters often differentiate at the border of patterning identities. Whether the information inherent in the intersection of patterning identities alone controls all aspects of neuronal cluster development (location, identity, and size) is unknown. Here, we focus on the cluster of the medial longitudinal fascicle (nMLF) and posterior commissure (nPC), located at the forebrain/midbrain (fore/mid) boundary, to address this issue. We first identify expression of the transcription factor Six3 as a common and distinct molecular signature of nMLF and nPC neurons in zebrafish, and we use this marker to monitor mechanisms controlling the location and number of nMLF/nPC neurons. We demonstrate that six3 expression is induced at the fore/mid boundary in pax2.1/no-isthmus and smoothened/slow muscle omitted mutants, where identities adjacent to the six3 cluster are altered; however, in these mutants, the subpopulation of six3-positive cells located within the mispatterned territory is reduced. These results show that induction of the six3 cluster is triggered by the information derived from the intersection in patterning identities alone, whereas correct cluster size depends, in a modular manner, on the identities themselves. The size of the six3 cluster is also controlled independently of neural tube patterning: we demonstrate that the prechordal plate (PCP) is impaired in mixer/bonnie and clyde mutants and that this phenotype secondarily results in an increased production of six3-positive cells at the fore/mid boundary, without correlatively affecting patterning in this area. Thus, a signaling process originating from the PCP distinguishes between neural patterning and the control of six3 cluster size at the fore/mid junction in vivo. Together, our results suggest that a combination of patterning related and -unrelated mechanisms specifically controls the size of individual early neuronal clusters within the anterior neural plate. PMID- 12889062 TI - Wnt11 and Wnt7a are up-regulated in association with differentiation of cardiac conduction cells in vitro and in vivo. AB - The heart beat is coordinated by a precisely timed sequence of action potentials propagated through cells of the conduction system. Previously, we have shown that conduction cells in the chick embryo are derived from multipotent, cardiomyogenic progenitors present in the looped, tubular heart. Moreover, analyses of heterogeneity within myocyte clones and cell birth dating have indicated that elaboration of the conduction system occurs by ongoing, localized recruitment from within this multipotent pool. In this study, we have focused on a potential role for Wnt signaling in development of the cardiac conduction system. Treatment of embryonic myocytes from chick with endothelin-1 (ET-1) has been shown to promote expression of markers of Purkinje fiber cells. By using this in vitro model, we find that Wnt11 are Wnt7a are up-regulated in association with ET-1 treatment. Moreover, in situ hybridization reveals expression, although not temporal coincidence of, Wnt11 and Wnt7a in specialized tissues in the developing heart in vivo. Specifically, whereas Wnt11 shows transient and prominent expression in central elements of the developing conduction system (e.g., the His bundle), relative increases in Wnt7a expression emerge at sites consistent with the location of peripheral conduction cells (e.g., subendocardial Purkinje fibers). The patterns of Wnt11 and Wnt7a expression observed in vitro and in the embryonic chick heart appear to be consistent with roles for these two Wnts in differentiation of cardiac conduction tissues. PMID- 12889063 TI - Induction of neurogenin-1 expression by sonic hedgehog: Its role in development of trigeminal sensory neurons. AB - We have examined the roles of signaling molecules in the mechanisms underlying the induction of neurogenin (ngn)-1 expression. ngn-1 is a basic helix-loop-helix (bHLH) transcription factor, which is essential for the specification of trigeminal sensory neurons. Semiquantitative reverse transcriptase-polymerase chain reaction using cranial explants in organ cultures showed that sonic hedgehog (Shh) promotes ngn-1 expression. This promoting activity was not observed in other signaling molecules examined. The promotion of ngn-1 expression by Shh, furthermore, was inhibited by cyclopamine, a specific inhibitor of Shh signaling. Shh did not affect the expression of ngn-2, a bHLH transcription factor that plays an important role in the specification of epibranchial placode derived sensory neurons. The expression levels of ngn-1 and ngn-2 decreased after fibroblast growth factor-2 treatment. These results suggest that Shh induces ngn 1 expression specifically and that expression of ngn-1 and ngn-2 is regulated by different mechanisms. The induction of ngn-1 expression by Shh suggests that this signaling molecule participates in the specification of trigeminal sensory neurons. We therefore examined the effect of Shh on the development of these neurons. Immunostaining using anti-ngn-1 demonstrated that Shh promotes ngn-1 expression in trigeminal neural crest cells. Trigeminal neural crest cells are derived from the posterior mesencephalon and the most-anterior rhombencephalon, and they contain a subset of precursors of trigeminal sensory neurons. Moreover, a subpopulation of trigeminal neural crest cells expressed the Shh receptor Patched. The number of cells that express Brn3a, a POU-domain transcription factor that plays an important role in differentiation of sensory neurons, also increased with Shh treatment. Our data suggest that Shh signaling is involved in the specification of trigeminal sensory neurons through the induction of ngn-1 expression. Furthermore, Shh promotes the differentiation of neural crest cells into trigeminal sensory neurons. PMID- 12889064 TI - Establishment of a ventral cell fate in the spinal cord. AB - The neural plate is induced during gastrulation when the organizer affects the ectoderm around it. Recent experiments show that axial mesoderm can stimulate formation of specific ventral cell types in the spinal cord, including floor plate, motor neurons, and several types of interneurons. We have eliminated or disrupted axial mesoderm by using a variety of methods to show that ventral columns of intermittent dopaminergic neurons in the frog Xenopus also appear to be induced by axial mesoderm. Inversion of the dorsal-ventral neural axis by splitting the presumptive neural plate in vivo, produced two spinal cords with ectopic dopaminergic neurons. The location and number of neurons suggest that even a brief association with axial mesoderm can specify the identity of the first or primary dopaminergic neurons and that notochord retains the ability to induce cells to become secondary dopaminergic neurons. PMID- 12889065 TI - Intercalary and supernumerary regeneration in the limbs of the frog, Xenopus laevis. AB - Anuran amphibians, such as Xenopus laevis, can regenerate their limbs only when they are young tadpoles, whereas urodele amphibians have a regenerative ability throughout their lives. It is still unclear whether anuran and urodele use the same mechanism during regeneration. In the present study, we analyzed intercalary and supernumerary regeneration in Xenopus. In contrast to urodele blastema that induces intercalary regeneration along the proximodistal (PD) axis, intercalation did not occur in the Xenopus limb bud when the presumptive zeugopodium (fibula and tibia) was removed. However, when the limb bud tip (presumptive autopodium) was transplanted to the presumptive stylopodium (femur) with a 180-degree rotation at stage 52, the complete zeugopodium was regenerated. These results were similar to the results of urodele mature limbs, suggesting that Xenopus limb buds are equivalent to the urodele mature limbs but not to the urodele blastemas. We hypothesized that the ability for intercalation depends on the expression pattern of fibroblast growth factor (fgf)-8, because the expression of fgf-8 in the urodele spreads over the whole blastema and is close enough to activate the growth of the stump. To test this hypothesis, an FGF-8-soaked bead was implanted at the boundary between the stump and tip of a Xenopus limb bud. Intercalary regeneration was induced at stages 52 and 53. These results suggest that the Xenopus limb bud possesses the potential for intercalation, but endogenous FGF-8 in the apical ectodermal ridge (AER) does not induce intercalation to the stump because of the long distance between the AER and stump. PMID- 12889066 TI - Developmentally regulated expression of Netrin-1 and -3 in the embryonic mouse molar tooth germ. AB - The Netrins form a small, conserved family of laminin-related signaling proteins regulating axon guidance in the developing nervous system. Here, we analyzed the roles of Netrin-1 and -3 in trigeminal axon guidance to the first lower molar of the embryonic mouse. Netrin-1 showed a restricted epithelial expression domain buccal to the tooth germ, toward which the pioneer tooth axons initially appear to navigate. Later, before birth, transcripts were colocalized with nerve fibers around the bell stage tooth germ. Analysis of Netrin-1-deficient mice, however, did not reveal any obvious disturbances in the axon growth or pattern of tooth innervation. In contrast, Netrin-3 showed a prominent, distinct expression in the axon pathway and target field mesenchyme around the tooth. Hence, it is possible that Netrin-3 may regulate pioneer axon growth toward and within the embryonic tooth target field. PMID- 12889067 TI - Embryonic stem cell and tissue-specific expression of a novel conserved gene, asrij. AB - We have identified a novel gene expressed in murine embryonic stem (ES) cells and in a restricted, tissue-specific pattern during mouse development. The gene is also expressed in blood vessels; hence, we have named it asrij (Sanskrit; asRij= blood). The gene encodes a novel conserved, predicted transmembrane protein of 247 amino acids, which is localized to lysosomes and endosomes. During ES cell derived blood vessel formation in vitro, Asrij expression precedes and partially overlaps with the vascular markers Flk-1 and PECAM. During development, Asrij is expressed predominantly in mouse embryonic blood vessels. The asrij transcript is alternatively spliced, and its expression is regulated in a tissue-specific manner. An asrij splice variant that is enriched in the adult mouse brain encodes a protein of 196 amino acids. Asrij can serve as an early stem cell marker that is down-regulated in nonvascular tissues. Our data indicate that Asrij belongs to a novel class of conserved proteins with a complex developmental profile and suggests multiple functions for the gene. PMID- 12889068 TI - Zebrafish atonal homologue zath3 is expressed during neurogenesis in embryonic development. AB - Basic helix-loop-helix (bHLH) transcriptional activators function in development of various cell lineages, including the central nervous system. One of the bHLH proteins, Math3/Xath3/NeuroM, was suggested to act as a late proneural gene in the mouse, Xenopus, and chick. Here, we isolated a zebrafish homologue, named zath3, and analyzed its expression pattern in zebrafish embryos. In the neural plate, zath3 is expressed first in the primordia of the tegmentum and trigeminal ganglia and three classes of primary neurons: sensory neurons, interneurons, and motor neurons. During later development, zath3 transcripts were localized along the boundaries of the optic tectum in the midbrain and rhombomeres of the hindbrain. Analyses of zath3 expression in three mid-hindbrain boundary mutants, acerebellar, no isthmus, and spiel-ohne-grensen, indicated that distribution of zath3 mRNAs in the midbrain and hindbrain was dramatically disturbed. In addition, these mutants also affect expression of zath3 in the neuroretina. PMID- 12889069 TI - Progranulin (acrogranin/PC cell-derived growth factor/granulin-epithelin precursor) is expressed in the placenta, epidermis, microvasculature, and brain during murine development. AB - The growth factor progranulin (acrogranin/PC-derived growth factor/granulin epithelin precursor) promotes onset of blastocyst cavitation and is required for neonatal hypothalamic sexual differentiation. Little is known, however, of the range of developmental processes in which it is involved. We used in situ hybridization to investigate progranulin expression in murine embryos. Progranulin mRNA is expressed in maternal and embryonic components during early establishment of pregnancy. Abundant expression is observed in the early decidualizing uterine stroma and glands. In the embryo, the trophoblast giant cells at the interface of placental exchange sites (both choriovitelline and chorioallantoic placenta) show strong expression. The gastrulating epiblast and mesenchyme (intraembryonic and extraembryonic mesenchyme) all revealed activity. The allantois and yolk sac mesenchyme (site of early hemopoiesis) were positive, as were later phases of active vessel formation (pia mater of brain, epicardium of the heart). In the urogenital system, it was expressed in Sertoli cells and in kidney tubules. It was highly expressed in proliferating epidermal cells. During epidermal appendage formation, the early epithelial bud was positive, but the forming duct and differentiating adjacent mesenchyme was negative. It is widely distributed during central nervous system development and the peripheral nervous system (dorsal root ganglia and sympathetic ganglia). Based on the pattern of progranulin gene expression, we propose proliferative and developmental roles for progranulin in establishing pregnancy, during gastrulation, and during embryonic development of the epidermis, nervous system, blood vessel, formation, and spermatogenesis. PMID- 12889070 TI - Candidate testis-determining gene, Maestro (Mro), encodes a novel HEAT repeat protein. AB - Mammalian sex determination depends on the presence or absence of SRY transcripts in the embryonic gonad. Expression of SRY initiates a pathway of gene expression resulting in testis development. Here, we describe a novel gene potentially functioning in this pathway using a cDNA microarray screen for genes exhibiting sexually dimorphic expression during murine gonad development. Maestro (Mro) transcripts are first detected in the developing male gonad before overt testis differentiation. By 12.5 days postcoitus (dpc), Mro transcription is restricted to the developing testis cords and its expression is not germ cell-dependent. No expression is observed in female gonads between 10.5 and 14.5 dpc. Maestro encodes a protein containing HEAT-like repeats that localizes to the nucleolus in cell transfection assays. Maestro maps to a region of mouse chromosome 18 containing a genetic modifier of XX sex reversal. We discuss the possible function of Maestro in light of these data. PMID- 12889071 TI - Expression of the Wdr9 gene and protein products during mouse development. AB - Human WDR9 has been mapped to chromosome 21, within one of the Down syndrome (DS) critical regions. Here, we study the expression pattern of the murine Wdr9 gene and its protein product. We show that Wdr9 is broadly expressed in the mouse embryo by means of in situ hybridization and immunohistochemistry. Wdr9 expression levels are dynamic during embryonic development as revealed by Northern blot analysis. We further show that WDR9 is a nuclear protein associated with BRG1, a SWI/SNF complex component. We also demonstrate that a polyglutamine containing region of the protein functions as a transcriptional activation domain. We propose that WDR9 is a transcriptional regulator involved in chromatin remodeling through the action of two bromodomains and contacts to the SWI/SNF complex. These results may provide a molecular basis for the association of WDR9 with DS. PMID- 12889073 TI - From functional neurosurgery to "interventional" neurology: survey of publications on thalamotomy, pallidotomy, and deep brain stimulation for Parkinson's disease from 1966 to 2001. AB - Articles on surgery for Parkinson's disease (PD), published between 1966 and 2001, were reviewed with respect to whether the first author had a neurosurgical affiliation, and whether the papers appeared in neurosurgical or non neurosurgical journals. Between 1966 and 1979, neurosurgeons and non neurosurgeons published almost equally on surgery for PD in both neurosurgical and non-neurosurgical journals; between 1980 and 1995, the majority of publications were by neurosurgeons in neurosurgical journals; and after 1995, non neurosurgeons were more often first authors of surgical publications and these were more frequent in non-neurosurgical journals. The fact that the first author of surgical publications on PD is often a non-neurosurgeon may have some bearing on the reported results of surgery. PMID- 12889074 TI - The story of penicillamine: a difficult birth. PMID- 12889075 TI - Spatial learning in rats is impaired after degeneration of the nigrostriatal dopaminergic system. AB - We investigated spatial learning in rats with unilateral and bilateral lesions of the nigrostriatal dopaminergic system. We used the Morris water maze paradigm, which tests spatial forms of learning and memory and allows discrimination between sensory-motor and learning disabilities. Animals were trained preoperatively to learn the location of a spatially fixed hidden platform to escape from the swimming pool (acquisition training). A visual and a probe test were used before and after the acquisition training, respectively. Our results show that animals with unilateral lesions, although displaying longer escape latencies, have normal spatial memory abilities. Animals with bilateral lesions were able to swim as fast or even faster than animals with unilateral lesion. Despite the fact that these animals had learned the spatial navigation tasks preoperatively, bilateral dopaminergic lesions led to a profound deficit in ability to find a hidden platform during an acquisition task. In general, animals with bilateral lesions persisted in swimming along the pool walls and their spatial navigation performance during a probe test was very poor. These results suggest that deficit of the nigrostriatal dopaminergic system can affect the selection and maintenance of behavioral strategies in spatial navigation. PMID- 12889076 TI - Fipamezole (JP-1730) is a potent alpha2 adrenergic receptor antagonist that reduces levodopa-induced dyskinesia in the MPTP-lesioned primate model of Parkinson's disease. AB - Previous studies in the MPTP-lesioned primate model of Parkinson's disease have demonstrated that alpha(2) adrenergic receptor antagonists such as idazoxan, rauwolscine, and yohimbine can alleviate L-dopa-induced dyskinesia and, in the case of idazoxan, enhance the duration of anti-parkinsonian action of L-dopa. Here we describe a novel alpha(2) antagonist, fipamezole (JP-1730), which has high affinity at human alpha(2A) (K(i), 9.2 nM), alpha(2B) (17 nM), and alpha(2C) (55 nM) receptors. In functional assays, the potent antagonist properties of JP 1730 were demonstrated by its ability to reduce adrenaline-induced (35)S GTPgammaS binding with K(B) values of 8.4 nM, 16 nM, 4.7 nM at human alpha(2A), alpha(2B), and alpha(2C) receptors, respectively. Assessment of the ability of JP 1730 to bind to a range of 30 other binding sites showed that JP-1730 also had moderate affinity at histamine H1 and H3 receptors and the serotonin (5-HT) transporter (IC(50) 100 nM to 1 microM). In the MPTP-lesioned marmoset, JP-1730 (10 mg/kg) significantly reduced L-dopa-induced dyskinesia without compromising the anti-parkinsonian action of L-dopa. The duration of action of the combination of L-dopa and JP-1730 (10 mg/kg) was 66% greater than that of L-dopa alone. These data suggest that JP-1730 is a potent alpha(2) adrenergic receptor antagonist with potential as an anti-dyskinetic agent in the treatment of Parkinson's disease. PMID- 12889077 TI - Extrapyramidal features in Parkinson's disease with and without dementia and dementia with Lewy bodies: A cross-sectional comparative study. AB - Risk factors predicting an increased risk of dementia in Parkinson's disease (PD) are not fully established. The dementia associated with PD (PDD) closely resembles dementia with Lewy bodies (DLB). Based upon a high frequency of non dopaminergic mediated clinical features in DLB, we predicted that a motor subtype comprising postural instability and balance problems would be more common in PDD. We examined extrapyramidal, cognitive, and affective features in 38 PD, 43 PDD, and 26 DLB patients in a cross-sectional study design. Motor subtype was subdivided into postural-instability gait difficulty (PIGD) or tremor (TD) dominant. The PIGD-subtype was more common in PDD (88% of cases) and DLB (69% of cases) groups compared with the PD group (38% of cases), in which TD and PIGD sub types were more equally represented (P < 0.001). Although the mean depression scores overall were modest, PDD patients scored significantly higher than PD, but not DLB patients (Cornell; P = 0.006, and Geriatric Depression scale, GDS-15; P = 0.001), while within the PD group, those patients with a PIGD subtype had greater depression scores than the TD subtype (GDS-15; P < 0.05). We conclude that non dopaminergic motor features are frequent in PDD. Neurodegeneration within the cholinergic system is likely to mediate many of these motor problems, as well as playing a significant role in determining the neuropsychiatric symptomatology of both PDD and DLB. PMID- 12889078 TI - Cardiac MIBG scintigraphy is a sensitive tool for detecting cardiac sympathetic denervation in Parkinson's disease. AB - [(123)I]Metaiodobenzylguanidine ([(123)I]MIBG) cardiac scintigraphy could be helpful to differentiate Parkinson's disease (PD) from multiple system atrophy (MSA), demonstrating that, in PD with autonomic failure but not in MSA, there is a myocardial postganglionic sympathetic dysfunction. To investigate whether this method is more sensitive than standard autonomic testing to detect early involvement of sympathetic cardiac efferent, we analyse MIBG myocardial uptake in 8 PD patients with normal autonomic testing (nondysautonomia PD group, NDPD) in comparison with 10 PD patients with abnormal autonomic testing (dysautonomia PD group, DPD) and 10 MSA patients. Global MIBG uptake was assessed using the ratio of [(123)I]MIBG uptake in the heart to the upper mediastinum (H/M) on planar scintigraphic data. Regional MIBG uptake was determined on two single photon emission tomography scans in regions of the left ventricle. The mean H/M ratios were significantly different among the three groups (P < 0.0001). H/M ratios of both NDPD and DPD patients groups (H/M = 1.83 +/- 0.50 and 1.24 +/- 0.40, respectively) were significantly lower than in MSA patients (H/M = 2.52 +/- 0.60). However, in NDPD patients, H/M was significantly higher than in DPD patients. When compared to MSA patients, NDPD patients showed a regional reduction in MIBG uptake in all left ventricle regions markedly in the apex and the inferior wall. Our results suggest that MIBG myocardial scintigraphy (analysis of both H/M ratio and regional MIBG uptake) may be more sensitive than standard autonomic testing for the early detection of silent autonomic dysfunction in PD. PMID- 12889079 TI - Cabergoline versus levodopa monotherapy: a decision analysis. AB - We evaluated the incremental cost-effectiveness of cabergoline compared with levodopa monotherapy in patients with early Parkinson's disease (PD) in the German healthcare system. The study design was based on cost-effectiveness analysis using a Markov model with a 10-year time horizon. Model input data was based on a clinical trial "Early Treatment of PD with Cabergoline" as well as on cost data of a German hospital/office-based PD network. Direct and indirect medical and nonmedical costs were included. Outcomes were costs, disease stage, cumulative complication incidence, and mortality. An annual discount rate of 5% was applied and the societal perspective was chosen. The target population included patients in Hoehn and Yahr Stages I to III. It was found that the occurrence of motor complications was significantly lower in patients on cabergoline monotherapy. For patients aged >/=60 years of age, cabergoline monotherapy was cost effective when considering costs per decreased UPDRS score. Each point decrease in the UPDRS (I-IV) resulted in costs of euro;1,031. Incremental costs per additional motor complication-free patient were euro;104,400 for patients <60 years of age and euro;57,900 for patients >/=60 years of age. In conclusion, this decision-analytic model calculation for PD was based almost entirely on clinical and observed data with a limited number of assumptions. Although costs were higher in patients on cabergoline, the corresponding cost-effectiveness ratio for cabergoline was at least as favourable as the ratios for many commonly accepted therapies. PMID- 12889080 TI - Head retraction reflex in stiff-man syndrome and related disorders. AB - The head retraction reflex (HRR) is a vestigial withdrawal reflex of the face and is suppressed in healthy subjects. We investigated the prevalence and electrophysiological patterns of the HRR in patients suffering from stiff-man syndrome (SMS, n = 28) and related disorders, stiff-limb syndrome (SLS, n = 2), and progressive encephalomyelitis with rigidity and myoclonus (PERM, n = 20). In patients with a positive HRR, the electromyographic (EMG) pattern was analyzed with surface recordings from the orbicularis oculi, sternocleidomastoid, trapezius, and the paraspinal extensor muscles at midthoracic and lumbar levels. On clinical testing, 17 of 28 SMS patients, 10 of 20 PERM patients, and 0 of 2 SLS patients had a positive HRR, ranging from a brief contraction of the neck extensors to violent retropulsion of the upper body. In all muscles, EMG reflex patterns elicited by gentle taps to the face or by electrical stimulation of the trigeminal nerve branches consisted of two components: an early, synchronous, brief burst with the shortest latency in trapezius (12.5-20.0 msec) and a late, asynchronous, longer response with latencies between 44 and 70 msec. We conclude that the HRR is an abnormal cutaneomuscular brainstem reflex that occurs in a considerable proportion of patients with SMS and related disorders. Although neither specific nor particularly sensitive for SMS, presence of the HRR in a setting with otherwise unexplained stiffness and spasms might help to avoid the otherwise frequent misdiagnosis of a psychogenic motor disturbance in patients with SMS. PMID- 12889081 TI - Lower risk of Parkinson's disease in an admixed population of European and Indian origins. AB - We studied whether the occurrence of Parkinson's disease (PD) in the Anglo Indians, an admixed population of European and Asian Indian origin, differs from Indians living in the same environment. Epidemiological studies show considerably higher prevalence of PD amongst white compared to non-white populations. Normal Indians contain a approximately 40% lower number of melanized nigral neurons compared to Caucasians from the UK. Anglo-Indians are an admixed population of European and Indian origin. We used the UK Parkinson's Disease Society Brain Bank clinical diagnostic criteria (steps 1 and 2) to diagnose PD in 84 of 493 residents (Indians, 409; Anglo-Indians, 84) living in elderly homes in Bangalore, India. Of these 84, 80 were Indians (19.5%) and 4 were Anglo-Indians (4.8%). Occurrence of PD is nearly five times higher amongst Indians compared to the Anglo-Indians (odds ratio, 3.9; 95% confidence interval, 1.3-12.9). We conclude that an admixture population of European and Indian origins, rather than averaging, might result in reduced occurrences of PD. Hence, studying an admixed population could provide crucial insights into understanding genetic mechanisms in the etiopathogenesis of PD. PMID- 12889082 TI - Mutation analysis of the parkin gene in Russian families with autosomal recessive juvenile parkinsonism. AB - Autosomal recessive juvenile parkinsonism (AR-JP) is a form of hereditary parkinsonism characterized by variable clinical presentations and caused by mutations in a novel gene, parkin, on chromosome 6q25.2-27. Until now, no Russian cases of parkin-associated AR-JP have been reported on. We recruited 16 patients from 11 Russian families with dopa-responsive movement disorders according to the following criteria: 1) family history compatible with autosomal recessive inheritance; 2) onset of symptoms at A). The majority of our parkin-associated cases were characterized by early onset dopa-responsive parkinsonism with benign course and slow progression (5 patients from two families have been followed for as long as 18-36 years), and 1 patient had a phenotype of dopa-responsive dystonia. This first description of Russian patients with AR-JP and molecularly proven parkin mutations confirms the widespread occurrence of this polymorphic hereditary extrapyramidal disorder. PMID- 12889083 TI - Altered corticomotor representation in patients with Parkinson's disease. AB - In 6 patients with Parkinson's disease (PD) and 6 age-matched controls, transcranial magnetic stimulation was applied at 56 regions over the motor cortex and premotor cortex of each hemisphere, with the first dorsal interosseous (FDI) muscle of both hands activated at 15% maximum voluntary contraction during stimulation. For each site, motor evoked potential (MEP) landmarks were recovered, including MEP amplitude, MEP onset latency, and silent period duration. Scaled MEP amplitudes were used to construct individual cortical maps of the FDI muscles. The maps revealed an anterior displacement of the muscle representation in PD patients. This anterior shift over motor cortical areas may reflect increased contributions of corticocortical connections between motor cortex and premotor cortical areas, possibly enhanced by the visual feedback aspect of the task. These alterations may reflect adaptations to the impairments in striatocortical circuits in PD. PMID- 12889084 TI - Transplantation in Parkinson's disease: PET changes correlate with the amount of grafted tissue. AB - We compared the striatal uptake of [(18)F]fluorodopa with [(76)Br]-FE-CBT, a positron emission tomography (PET) ligand of the dopamine transporter (DAT), which estimates the density of dopamine nerve terminals, in 6 patients with Parkinson's disease grafted with fetal mesencephalic cells. There was no change in DAT ligand binding in the grafted putamen, despite a significant increase of [(18)F]fluorodopa uptake. This finding suggests that the clinical benefit induced by the graft is more related to increased dopaminergic activity than improved dopaminergic innervation in the host striatum and, therefore, that [(18)F]fluorodopa remains the optimal tracer to evaluate grafted PD patients. Further analysis showed that the clinical and [(18)F]fluorodopa uptake changes after the grafts were correlated with the number of ventral mesencephalae used for implantation. PMID- 12889085 TI - Staged lesions through implanted deep brain stimulating electrodes: a new surgical procedure for treating tremor or dyskinesias. AB - Thalamotomy and pallidotomy have been shown to have some efficacy for treating some movement disorders such as disabling tremor or parkinsonian levodopa-induced dyskinesias (LID). Compared to continuous deep brain stimulation (DBS), this surgical procedure has the disadvantage of irreversibility and a lack of adaptability. Making a lesion involves a risk of inducing permanent side effects, especially if the lesion is large, or of observing a resurgence of the symptoms if the lesion is too small. We performed unilateral pallidotomy in one patient suffering from LID and unilateral thalamotomy in two patients suffering from tremor through the lead classically used for DBS. The technique of lead implantation was similar to that used for DBS treatment but, instead of connecting the lead to a pulse generator, it was left in place and used to make a radiofrequency lesion. This technique allowed the lesion to be kept as small as possible, thereby minimizing the risk of permanent side effects and made possible to extend the lesion if the symptoms reappeared. One lesioning session was enough to relieve tremor in the two patients treated by thalamotomy; three lesioning sessions over a 7-month period were required to relieve drug-induced dyskinesias in the patient treated by pallidotomy. In all 3 patients, disabling symptoms were still relieved without any permanent side effects 6 months after the last lesion was performed. PMID- 12889086 TI - Olfactory bulb in multiple system atrophy. AB - Olfactory dysfunction is a characteristic clinical sign in Parkinson's disease (PD); it is also present in multiple system atrophy (MSA). The pathological basis of hyposmia or anosmia in PD is well known: the olfactory bulb (OB) contains numerous Lewy bodies and severe neuronal loss is present in the anterior olfactory nucleus (AON). We established that glial cytoplasmic inclusions (GCIs) are present in all the OBs from MSA cases. Their presence in the OB is diagnostic for MSA. Additionally, neuronal loss is present in the AON in MSA. These pathological changes might be responsible for the olfactory dysfunction seen in MSA. PMID- 12889087 TI - Unique form of propriospinal myoclonus as a possible complication of an enteropathogenic toxin. AB - Propriospinal myoclonus is an uncommon form of spinal myoclonus propagated, presumably, by slowly conducting polysynaptic intraspinal pathways. Although most patients demonstrate no clear etiology, a variety of disorders have been linked to this abnormal movement, including trauma, multiple sclerosis, tumors, and infectious disorders such as herpes zoster, human immunodeficiency virus, and Lyme disease. We describe 2 young male patients from the same town in Northern Ontario, Canada, exposed to an outbreak of Escherichia coli O157:H7 from contaminated municipal water, who developed identical clinical and electrophysiological features suggestive of a rhythmic form of propriospinal myoclonus with activity alternating between abdominal and paraspinal muscles. A toxin-mediated microvascular thrombosis is proposed as a possible pathogenic mechanism underlying this novel association. PMID- 12889088 TI - Multiple sclerosis tremor and the Stewart-Holmes manoeuvre. AB - The objective of this study is to define tremor and cerebellar dysfunction and determine whether kinetic and postural tremor correlate with cerebellar dysfunction in patients with multiple sclerosis (MS). Cerebellar symptoms such as dysmetria often interfere with tremor evaluation in MS. The Stewart-Holmes (SH) manoeuvre, which has been recently quantified, may offer a selective evaluation of cerebellar dysfunction in such patients. Thirty-two patients with definite MS and arm tremor were evaluated (simplified Fahn tremor scale for kinetic and postural tremor, finger-to-nose test, clinical SH manoeuvre, quantitative study of the SH manoeuvre). Median severity of kinetic and postural tremor on the most disabled side was, respectively, 2 (range 0-4) and 1.5 (range 0-4). Clinical SH scores were moderately correlated to quantified SH measures (r = 0.36, P < 0.05). Kinetic and postural tremors were strongly correlated (r = 0.73, P < 0.0001) but did not correlate with clinical or quantified SH scores. Patients with bilateral tremor had higher scores for quantified SH, and a trend to higher clinical SH and finger-to-nose scores than patients with unilateral tremor. Although clinically associated, cerebellar dysfunction and tremor may be partly independent symptoms, suggesting they may relate to dysfunction of different neuronal systems. The SH manoeuvre should be part of the evaluation of MS patients considered for surgery of tremor. PMID- 12889089 TI - Influence of monocyte chemoattractant protein 1 gene polymorphism on age at onset of sporadic Parkinson's disease. AB - We studied polymorphisms in the genes for monocyte chemoattractant protein 1 (MCP 1) and CC chemokine receptor (CCR)-2 in 171 Parkinson's disease (PD) patients and 340 controls. Although no associations were found in alleles or genotypes, MCP-1 2518A/G genotype affected the age-at-onset of PD patients. This effect was also detected in a second PD group, suggesting a possible involvement of MCP-1 in PD. PMID- 12889090 TI - Delineating the sites and progression of in vivo atrophy in multiple system atrophy using fluid-registered MRI. AB - We describe the pattern and progression of atrophy delineated using fluid registration of serial magnetic resonance imaging scans in a case of multiple system atrophy (MSA). The in vivo findings were consistent with those found at postmortem, including significant supratentorial atrophy concurrent with an unusual degree of cognitive impairment for MSA. PMID- 12889091 TI - Life-threatening respiratory failure due to cranial dystonia after dental procedure in a patient with multiple system atrophy. AB - We report on a woman with a an 8-year history of multiple system atrophy with predominance of parkinsonism who developed jaw-locking oromandibular dystonia within hours after insertion of ill-fitting dentures. Dystonia spread rapidly to involve other facial muscles and the larynx causing stridor with respiratory failure necessitating crush intubation. PMID- 12889092 TI - Myoclonus in spinal dysraphism. AB - Two cases of segmental myoclonus occurring in association with spinal dysraphism are described. In one, myoclonus of paralysed legs arose below a region of spinal cord lacking any normal function, illustrating the capacity of the isolated spinal cord to generate and maintain rhythmic activity independent of supraspinal influences. PMID- 12889093 TI - Abstracts of the International Continence Society 33rd annual meeting. Florence, Italy, 5-9 October 2003. PMID- 12889094 TI - After-effects of head injury. PMID- 12889095 TI - Sarcoidosis. AB - The historical background to the disorder is unfolded. It was originally regarded as a dermatological curiosity, and later sarcoidosis was recognized as a disorder diffusely involving most tissues of the body. Clinical syndromes described include those presenting to the dermatologist, ophthalmologist, chest physician and radiologist, gastroenterologist and neurologist. Tissues commonly involved are lungs, lymph nodes, eyes, skin and bone in that order of frequency. Granulomatous uveitis in the presence of skin lesions should always arouse the suspicion of sarcoidosis; ocular and bone involvement are frequently associated with lupus pernio, whereas lymphadenopathy and splenomegaly are more commonly linked with plaques and maculo-papular eruptions. Bone cysts are rare in the absence of skin lesions, so routine radiography of hands and feet is of little diagnostic value. The basic criteria for establishing the diagnosis of sarcoidosis are twofold: (a) Suggestive clinical and/or radiological features with evidence of generalized involvement. (b) Histological proof of sarcoid tissue from at least one tissue. Evidence of one without the other is insufficient, for clinical or radiological manifestations alone present too wide a differential diagnostic problem and, conversely, isolated histological evidence of sarcoid tissue could be construed as a local sarcoid-tissue reaction. Histological confirmation is obtained whenever possible from accessible involved tissues (skin or lymph node). Otherwise blind biopsy of liver, scalene lymph node or gastrocnemius muscle is employed, or alternatively the Siltzbach-Kveim test is performed. It is a safe, simple and specific outpatient skin test, which provides histological confirmation in three-quarters of patients with sarcoidosis. The natural history of sarcoidosis and the response to treatment vary with the type of disease--namely subacute (transient) or chronic (persistent) forms of sarcoidosis. The only treatment which favourably influences clinical, or radiological or histological features of the disease is corticosteroid therapy. Indications for treatment are: ophthalmic involvement, steadily worsening chest radiograph, breathlessness, persistent hypercalciuria, disfiguring skin lesions, neurological involvement, disordered glandular function. PMID- 12889096 TI - Carpal tunnel syndrome. PMID- 12889097 TI - The investigation of vaginal discharge in women and children. PMID- 12889098 TI - Membrane transport of drugs. PMID- 12889099 TI - [Protein kinase C in eosinophil proliferation and infiltration in nasal polyp tissue]. AB - OBJECTIVE: To explore the significance of protein kinase C (PKC) in eosinophil (EOS) proliferation and infiltration in nasal polyp tissues. METHODS: With the methods of in situ hybridization and immunohistochemistry staining, PKC and apoptosis gene (Bcl-2, Bax) expression were measured between nasal polyps from 26 patients and inferior turbinate mucous membrane tissue (ITMMT) from 20 normal persons. EOS was identified with May-Grunwald-Giemsa (MGG) staining. RESULTS: In EOS, the positive cell expressive rate of Bcl-2 mRNA and its protein were significantly higher in group of nasal polyps than that in group of ITMMT (P < 0.01). Although the positive cell expressive rate of Bax mRNA and its protein were slightly higher in group of nasal polyp tissue than that in group of ITMMT, the difference was not significant (P > 0.05). PKC was expressed in EOS of all 26 cases from nasal polyp tissues, but in 20 cases from ITMMT, only 7 cases showed occasional PKC expression. In both groups, PKC positive cell expression was significantly different, and the expression of PKC and Bcl-2 mRNA as well as their protein in EOS of nasal polyps showed remarkably positive relation (r1 = 0.0875, r2 = 0.0823, P < 0.01). CONCLUSION: PKC expression increasing in EOS of nasal polyp tissues was closely associated with apoptosis inhibiting. So we presumed that EOS inhibiting in nasal polyp tissues was obtained by activating PKC signal transduction pathway. PMID- 12889100 TI - [Expression of chloride channel in nasal mucosa with allergic rhinitis]. AB - OBJECTIVE: To evaluate the role of chloride channel ClC-3 in perennial allergic rhinitis. METHODS: The nasal mucosa from 19 patients with perennial allergic rhinitis were studied by immunohistochemistric staining and reverse transcriptase polymerase chain reaction (RT-PCR) for the expression of ClC-3. RESULTS: ClC-3 was positively stained in 17 cases of 19 patients, with a rate of 89.6%. It mainly distributed in cytoplasm of epithelium and submucosal gland. All tissues displayed expected DNA length after PCR. The relative density of ClC-3 in mucosa with allergic rhinitis was 0.685 +/- 0.114 and 0.140 +/- 0.075 in normal. CONCLUSION: ClC-3 played an important role in the over-secretion of liquor in allergic rhinitis. PMID- 12889101 TI - [Study of pathological morphology in chronic sinusitis]. AB - OBJECTIVE: To study the pathological characteristics and regularity of chronic sinusitis (CS) in different clinical types, tc estimate the reliability and feasibility of CS classification on the pathologic basis. METHODS: HE, Van Gieson's, alcian blue-periodic acid-schiff(AB-PAS) staining was used to observe the changes of epithelium, to measure the thickness of basement membrane and the area of mucous glands. The ultrastructural changes of pseudostratified ciliated columnar epithelium were observed under transmission electron microscope (TEM) and scanning electron microscope (SEM). RESULTS: The epithelial fiber tissue was increased, the basement membrane was thickened, and the mucous glands were obviously increased in CS under the light microscope. Subepithelial fibrosis and thickened basement membrane were most obviously seen in type III of CS and the hyperplasia of mucous glands was most distinct in type II. Under the TEM, abnormal structure of cilia, decrease of ciliated cells, increase of goblet cells were detected in all three CS types. Under the SEM, the cilia of every types showed the ingravescenced process of pathological changes from slight cilium disorder, disorientation, to desquamation of most cilia and replacement of microvilli. CONCLUSION: There is significant distinction of mucous membrane structure among three types. The Chinese classification of CS has pathological basis, it represents the occurrence, development and regularity of CS, and it can be used as basis in further study, communication and evaluation for surgical effect of CS. PMID- 12889102 TI - [Detection of eosinophil cationic protein and E-selectin in serum and nasal lavage fluid in patients with allergic rhinitis]. AB - OBJECTIVE: To evaluate the role of eosinophil cationic protein (ECP) and serum soluble E-selectin (sE-selectin) in patients with allergic rhinitis. METHODS: The sE-selectin were detected from 30 allergic patients and 20 normal individuals by enzyme-linked immunosorbent assay (ELISA). The ECP in serum and nasal lavage fluid was assayed with pharmacia CAP system. RESULTS: The level of sE-selectin in patients with allergic rhinitis was(41.846 +/- 12.21) ng/ml, higher than that in control (10.678 +/- 2.458) ng/ml. The level of ECP in serum and nasal lavage fluid from patients with allergic rhinitis was significantly higher than that from normal controls. The level of ECP in nasal lavage fluid was much higher than that in serum. CONCLUSION: ECP and sE-selectin play a role in pathogenesis of allergic rhinitis. PMID- 12889103 TI - [Clinical evaluation of local application of Merogel after endoscopic sinus surgery]. AB - OBJECTIVE: To study the auxo-action of Merogel (hyaluronic acid) for operative cavity epithelialization after endoscopic sinus surgery (ESS). METHOD: Sixteen patients (32 sides) were observed, in which all sinuses(maxillary, frontal, ethmoid and sphenoid) were operated on by ESS. For the purpose of control study, the left nasal side was used as Merogel group, and right side contrast group. The patients were followed-up for 12 weeks. RESULTS: In Merogel group, the curative rate was 93.75%, improvement rate 6.25%, and average time of epithelialization was 3.4 weeks. In contrast group, the curative rate was 87.50%, improvement rate 12.50%, and average time of epithelialization was 8.3 weeks. It was distinct that in Merogel group, the epithelialization time was shorter than that in contrast group. CONCLUSION: Merogel has auxo-action for operative cavity epithelialization after ESS. PMID- 12889104 TI - [Cisplatin-induced apoptotic cell death in spiral ganglion and organ of Corti of mongolian gerbil cochlear]. AB - OBJECTIVE: To investigate whether apoptosis is one of the mechanism for cell death in spiral ganglion(SG) and organ of Corti of Mongolian gerbils cochlea induced by cisplatin. METHODS: Mongolian gerbils were administered 4 mg.kg-1.d-1 cisplatin consecutively for 4 to 7 days. The apoptotic cell death in spiral ganglion and organ of Corti of cochlea basal turn was detected via transmission electron microscopy and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) method. RESULTS: The observation of electron microscopy shown special morphological change consistent with the feature of cell apoptosis in some spiral ganglion cells and outer hair cells after 5-7 days' cisplatin treatment. The apoptotic labeling by TUNEL in spiral ganglion and organ of Corti was detected after 5 days' treatment and increased after 6 and 7 days' treatment whereas almost negative result were obtained in the normal control animals. CONCLUSION: Apoptosis is one of the important mechanisms for cell injury of spiral ganglion and organ of Corti induced by cisplatin. PMID- 12889105 TI - [Change of eustachian tube surfactant of guinea pigs suffering from secretory otitis media]. AB - OBJECTIVE: To observe the change of surface active agents of eustachian tube of guinea pigs suffering from secretory otitis media, to explore the effects of surface active agents on secretory otitis media. METHODS: To establish the animal model of guinea pigs suffering from secretory otitis media. To analyse the changes of biochemical component and activity of eustachian tube's surfactant, meanwhile; to observe surfactant change after ectogenic surfactant treatment. RESULTS: The main components of the surfactant decrease in the model group of guinea pigs suffering from secretory otitismedia, phosphatidylcholine (PC) is (19.9 +/- 1.7)%, phosphatidylethanolamine(PE) is (36.8 +/- 2.7)%, minimum surface tension (gamma min) is (18.5 +/- 2.4) mN/m, and PC is (25.7 +/- 2.1)%, PE is (43.7 +/- 3.8)%, (P < 0.001), and gamma min is (7.6 +/- 0.8) mN/m in normal group (P < 0.001). After giving Ectogenic surfactant treatment, PC and PE increase, PC is (23.3 +/- 2.2)%, PE is (42.5 +/- 3.6)%; gamma min reduce, it is (11.8 +/- 2.3) mN/m. CONCLUSIONS: The main biochemical components of the surfactant obviously decrease, the activity of the surfactant weakens in secretory otitis media of guinea pig. This study showed the treatment of Ectogenic surfactant is effective. PMID- 12889106 TI - [Implication of cochlear implantation in patients with enlarged vestibular aqueduct syndrome]. AB - OBJECTIVE: To evaluate the intraoperative obstacles and post-operative outcomes of cochlear implantation in children with enlarged vestibular aqueduct syndrome. METHODS: Between May 1995 and June 2002, 10 patients with enlarged vestibular aqueduct syndrome received cochlear implantation in Pekin Union Medical College Hospital. The clinical records of 10 cases of enlarged vestibular aqueduct syndrome were analyzed retrospectively. RESULTS: Via cochleostomy pulsatile clear fluid gusher occurred in 8 cases, which were easily controlled by quickly inserting the electrode array and sealing the cochleostomy with muscle tissue in each instance. The implant was inserted without difficulty in all patients. There have been no post-operatively complications, and audiologic outcome of the 10 patients with cochlear implantation were almost the same as the other patients without inner ear malformations. Five cases of them can even speak better than the other prelingual deafness that may be attributed in part to postlingual deafness. CONCLUSIONS: Despite pulsatile clear fluid gusher in cochlear implantation, no post-operatively complications have been found, and the outcome of these 10 patients with cochlear implantation was excellent. Cochlear implantation can be safely and effectively performed in patients with profound sensorineural hearing loss (SNHL) associated with enlarged vestibular aqueduct syndrome. PMID- 12889107 TI - [Cochlear implants in children with enlargement of vestibular aqueduct]. AB - OBJECTIVE: To describe the clinical experiences with multi-channel cochlear implantation in the children with enlargement of vestibular aqueduct. METHOD: Fifteen patients were diagnosed with enlargement of vestibular aqueduct and received multi-channel cochlear implantation at Beijing Tongren Hospital. A cochleostomy was performed for all patients by a standard facial recess approach to anterior inferior to the round window niche. RESULTS: (1) Most of the electrodes were totally inserted in the cochlear, and gusher appeared in 6 implantations. (2) No serious complications occurred after implantation. (3) All patients have auditory sensations after implantations. (4) The hearing threshold in this series were similar to that of the normal cochlear implantation (the average threshold was 30 dB). (5) The rehabilitation concept was developed by Rehabilitation Research Center for Deaf Children in Beijing and Beijing Tongren Hospital. The training sessions included sound detection, word discrimination, auditory comprehension and spoken language skill development. The abilities of speech discrimination and spoken language were improved comparing with that of pre-operation. Two of the patients could go to the normal school after implantations. CONCLUSION: The outcome of hearing rehabilitation for children with enlargement of vestibular aqueduct were similar to those children with "normal" cochlear structure followed the multi-channel cochlear implantation. PMID- 12889108 TI - [Endoscopic technique utilized into the selective vestibular neurotomy by retrosigmoid approach]. AB - OBJECTIVE: To investigate the micro-wound technique and the signification of reducing brain tissue oppression and complication at the surgery with utilizing endoscopic technique during the operation of selective vestibular neurotomy (SVNT) by retrosigmoid approach. METHODS: Twelve patients were preceded the SVNT by retrosigmoid approach (as routine operation group, ROG), and nine patients were utilized endoscope for review and neurotomy during the process of the SVNT by retrosigmoid approach (as endoscopic operation group, EOG). All of them were examined by auditory level, vestibular function, facial nerve function and observed of clinical situation (including complication and equilibrium function and so on) before and after operation. RESULTS: Two patients appeared cerebral edema and demanded the treatment of dehydration and depressing intracranial pressure in ROG after operation. In each group, the average auditory threshold in 2 patients rose more than 15 dB post-operation. The interval time of average equilibrium function compensation is (29.00 +/- 9.60) d in ROG and (28.56 +/- 7.91) d in EOG after surgery. Peripheral facial paralysis didn't happen to any patient in two groups. After follow-up two years post-operation, the vertigo recurred in 2 patients (4 times and 1 times, respectively) in ROG and 1 patient (2 times) in EOG. CONCLUSIONS: The SVNT by retrosigmoid approach is widespread adopted presently as expedience and safe surgery method for Meniere's disease, and could effectively eliminate vertigo and simultaneously conserve the hearing function and the integration of facial nerve. This surgery process became micro wound, more facility and safe following the endoscopic technique utilization, which could reduce the brain tissues depression and trauma and could not cause any of complications. Therefore, the endoscopic operation of the SVNT by retrosigmoid approach is worth to clinically popularize as the therapy method of vertigo for the patients of Meniere's disease, especially who associate with contralateral sensorineural hearing loss. PMID- 12889109 TI - [Prevention and therapy of cerebrospinal fluid leakage after translabyrinthine acoustic neuroma removal]. AB - OBJECTIVE: To evaluate the modified wound closure technique and other methods for preventing and treating the cerebrospinal fluid (CSF) leakage after removal of acoustic neuroma by translabyrinthine approach. METHODS: In a series of 85 cases of acoustic neuroma surgery, two kinds of technique of wound closure, the traditional (as first group) and the modified (as second group), have been subsequently used to prevent CSF leakage. The incidences of CSF leak were analyzed respectively. Conservative and surgical treatments were applied to treatment the CSF leak. RESULTS: In the first group, the CSF leak was 19.5% (8/41), and that of second group was 2.3% (1/44). There was significant difference between them (P = 0.013). The CSF leak mainly occurred in large acoustic neuromas surgery with traditional wound closure technique. Among 9 cases of CSF leaks, 3 were controlled by conservative method, 5 by one single revision surgery and one by two revisions. CONCLUSIONS: The revision technique of wound closure could significantly reduce the incidence of CSF leakage after acoustic neuroma removal by translabyrinthine approach. The revision surgery is an effective method for stopping the CSF leak. PMID- 12889110 TI - [Clinical study on applying brainstem auditory evoked potential to monitor anesthesia depth and awaken in children]. AB - OBJECTIVES: To study applying auditory brainstem reaction (ABR) to monitor the general anesthesia depth and awaken degree in children as an objective guide. METHODS: Forty-five infant or children patients, American Society of Anesthesiologists(ASA) graded I-II, with normal audition, were selected and divided into 3 groups (15 cases per group) to receive propofal venous anesthesia, fentanyl venous anesthesia, or isoflurane anesthesia, respectively. During anesthesia, peak latencies (PL) of I, III, V wave, and interpeak latencies (IPL) of I-III, III-V, I-V were monitored and recorded after short sound stimulation of 90 db which accumulated 1,000 times with Madsen. The changing of PL and IPL with time passing and anesthesia dosage was studied to discuss the feature of ABR in each anesthesia procedure as above. RESULTS: There is a significant positive correlation between PL and IPL of ABR waves as above and the dosage of propofol or the concentration of isoflurane. The dosage or the concentration increased, and PL and IPL prolonged. It is important of PL of wave I to reflect anesthesia degree. It is wave V with the best stability and correlation to monitor anesthesia degree. After anesthesia period or patients almost awaken, PL and IPL of some ABR waves were still more than normal values, which is detention reaction. For the fentanyl anesthesia, ABR is not sensible enough, i.e. has little correlation. CONCLUSION: The changing of PL and IPL of ABR waves could conclude anesthesia degree in children and be treated as reference of whether awaken or not in some certain, but have possibility of detention reaction. PMID- 12889111 TI - [Resection of malignant tumors of the anterior skull base and the fronto-orbito ethmoidal region by overturn of frontonasal flap]. AB - OBJECTIVE: To introduce a better surgical approach for the resection of malignant tumors in the anterior skull base and in the fronto-orbito-ethmoidal region. METHODS: A "U" form incision on the bilateral superciliary arch was made and connected with the unhealthy nasal side incision. Then the frontal musculocutaneous flap was over-turned upward. The nasal pyramid was over-turned to the healthy side. The malignant tumors in the anterior skull base and the fronto-orbito-ethmoidal region were fully exposed and resected. RESULTS: From december 1986 to december 1996, 50 patients received tumour resection through this approach. Postoperatively, the one-year survival rate was 86.0% (43/50), the three-year survival rate was 62.0% (31/50), and the five-year survival rate was 44.0% (22/50). CONCLUSION: The frontonasal flap overturn approach provided good exposure of surgical field, avoided the coronal incision, and the reaction of brain was mild. We believe this surgical approach is superior to traditional combined craniofacial approach. PMID- 12889112 TI - [Efficacy of selective neck dissection: a review of 123 cases of elective and therapeutic treatment of the neck in squamous cell carcinoma of head and neck]. AB - OBJECTIVE: To evaluate the efficacy of selective neck dissection (SND) in elective and therapeutic treatment of the neck in squamous cell carcinoma of head and neck. METHODS: A retrospective review was undertaken of 123 patients undergoing 157 SNDs as a part of initial therapy for squamous cell carcinoma of head and neck from January 1997 to September 2001 at this institute. The primary site included larynx(n = 77), oral cavity(n = 29), oropharynx(n = 2), and hypopharynx(n = 15). Lymph nodes were pathologically negative (pN0) in 99 and positive (pN+) in 24 patients. Peri-operative radiotherapy was given to 30.3% of pN0 and 50.0% of pN+ patients. The median follow-up interval was 25 months. RESULTS: 101 patients received elective neck dissection, 14 of them (13.9%) found to be pN+, while 22 patients received therapeutic neck dissection, 10 of them (45.5%) found to be pN+. A total of 52 positive nodes were found in 157 SNDs. The distribution of the positive nodes were as follows: Level I 25%; Level II 48%; Level III 25%; Level IV 2%. The 5-year regional recurrence rates estimated according to Kaplan-Meier were 5.87% (95% CI: 0.8%, 10.9%) for the pN0 and 9.2% (95% CI: 0.0%, 21.5%) for pN+ patients. CONCLUSION: The SND, when carefully indicated, a contribution to the concept of less invasive surgery, offers functional and esthetic advantages without oncologic compromise. PMID- 12889113 TI - [Simultaneous double free flap transfer for extensive defects in head and neck region: a review of 33 consecutive cases]. AB - OBJECTIVE: To analyze the rationale and feasibility of double free flap transfers for reconstruction of extensive head and neck defects. METHODS: Thirty-three consecutive cases of double free flap transfers for extensive head and neck defects from March 2000 to January 2002 were reviewed. Data concerning the operation included date of surgery, defect description and site, choice and combination of flap, recipient vessel and complications. RESULTS: The fibula osteocutaneous-radial forearm fasciocutaneous flap combination was most commonly used (n = 23), followed by the fibula osteocutaneous-rectus abdominis myocutaneous flap (n = 6) and radial forearm-rectus abdominis myocutaneous flap(n = 4). The complete flap survival rate was 97.0% (64 of 66 flaps). CONCLUSION: In selected cases, the double free flap procedure for reconstruction of massive head and neck defect is justified and effective, and greatly improve the quality of life for these patients. PMID- 12889114 TI - [Superior mediastinum exposure in the removal of the advanced cancers in the lower neck regions]. AB - OBJECTIVE: In order to achieve the radical removal of advanced cancers involving the lower neck regions. METHODS: Upon the thorough evaluation of tumors with different imaging methods, several kinds of superior mediastinum exposure were carried out in a series of 18 patients with advanced neck cancer of various kinds. There were 10 cases of stomal recurrence, 2 cases of recurrent thyroid carcinomas, 2 cases of advanced carcinomas in the cervical trachea, and 4 cases of metastatic carcinomas in the supraclavicular region. According to the site and extension of tumor invasion, simple manubrium resection were performed in 10 cases, resection of the medial half of the clavicle was adopted in 4 cases and resection of the manubrium and the medial 1/3 of both clavicles were used in 4 cases. Pectoralis major myocutaneous flaps were raised to reconstruct the defects resulting from tumor ablations. RESULTS: Radical removal of tumors was achieved in 16 cases. Minor complications occurred in 3 cases including one case of intraoperative pneumothorax and 2 cases of postoperative pharyngeal fistula. One patient developed aneurysm in the right common carotid artery after radical neck dissection and wound infection. One patient died of eruption of the innominate artery due to the major vessel injury caused by the tracheostomy tube two months after the operation. Results from postoperative follow-up demonstrated that one year, two-year and three-year postoperative survival rate was 72.2% (13/18), 22.2% (4/18) and 11.1% (2/18), respectively, in this series. CONCLUSION: Superior mediastinum exposure is a necessary and reliable surgical approach in the removal of advanced tumors involving the lower neck and superior mediastinum. PMID- 12889115 TI - [Clinical observation and histological research of vocal cord's reformation after laser cordectomy]. AB - OBJECTIVE: Explore the reformation of new vocal cord after laser cordectomy and the histological change of new vocal cord by animal experiment. METHODS: The reforming course of new vocal cord was observed by follow-up. 4 dogs underwent bilateral cordectomy (type II operation) and their larynx were analyzed anatomically and histologically eight months later. 100 cases who underwent KTP laser cordectomy were divided into 3 types according to the range and depth of the excision. RESULTS: All the dogs had reformation of vocal cords eight months later. On the surface of the reformed vocal cord, no mucous cell but squamous cells was found. Newborn ligament-like connective tissues and loose muscle fibers were found under membrane. Complete reconstruction of larynx was found in 16 cases of type I (16/16) and 54 of type II 86% (54/63). But in type III no new vocal cord was found. CONCLUSION: Formation of the new vocal cord is the result of hypertrophy of the remained vocal cord muscle, influenced by depth of the excised vocal cord using laser. The histological construction is nearly normal cord. Whether or not the laser radiation stimulate the tissue hypertrophy is need more research. PMID- 12889116 TI - [Correlation between prognosis and the expression of eukaryotic translation initiation factor 4E in surgical margin specimen of laryngeal squamous cell carcinoma]. AB - OBJECTIVE: To reveal the correlation between prognosis and the expression of eIF4E in the pathological negative surgical margin patients of laryngeal squamous cell carcinoma (SCC). METHODS: Immunohistochemistry was used to detect the expression of eIF4E protein in the pathological negative margin specimen and primary site of cancer of 67 patients, and 5 year survival rate data of those patients was analyzed retrospectively. RESULTS: The positive expression rate of eIF4E protein in surgical margin and primary site of cancer was 32.8% (22/67) and 100% (67/67) respectively. As for the control group, in which the eIF4E protein expression was not be found. The local recurrence rate was 63.6% for patients with eIF4E positive margin and 28.9% for patients with eIF4E negative margin (P = 0.006). The 5-year survival rate of patients with eIF4E positive margin was lower than those with negative margin (43.31% vs 77.52%, P = 0.0006). CONCLUSION: The prognosis of patients with eIF4E protein positive expression in the pathological negative surgical margin may be worse than those of the negative. PMID- 12889117 TI - [Tuberculosis of pharynx and larynx: a report of 32 cases]. AB - OBJECTIVE: To evaluate the trends and the clinical changes in tuberculosis of pharynx and larynx. METHODS: The clinical data of 32 patients with tuberculosis of pharynx and larynx from Jan. 1982 to Dec. 2000 in Daping hospital were studied retrospectively. RESULTS: (1) The local manifestations were mainly single lesion that commonly involved the vocal cord (10 cases). (2) The lesions appearances were mainly the proliferation such as mass (11 cases) or granulation(8 cases). (3) anti-tuberculosis is the main treatment, the operation is the second. Twelve patients cured in clinic, six patients received operation and cured without any complications. Fourteen patients condition controlled. CONCLUSION: The classical manifestations with tuberculosis of pharynx and larynx were not exited, the new clinical manifestations were associated with local lesion in nowadays. PMID- 12889118 TI - [Evaluation of laryngotracheal stenosis with spiral CT three-dimensional images and clinical application]. AB - OBJECTIVE: To study the use of spiral computed tomography three-dimensional images (SCT-3DI) in the evaluation of patients with laryngotracheal stenosis. The value and utility of SCT-3DI compared with actual endoscopy were also investigated in terms of accuracy of diagnosis and validity of treatment. METHODS: Eight patients with laryngotracheal stenosis were underwent SCT-3DI and regular endoscopy examination preoperatively. Of eight patients, Five had traumatic causes, two with tracheal narrowing due to tracheal granulomas, one with too much scar tissue of unknown etiology in the subglottic area. The location, extent and contour of the stenosis were identified and compared with actual endoscopy findings, and confirmed by surgical procedures. RESULTS: SCT-3DI evaluation was accurate in assessing the length, width and shape of all the laryngotracheal stenosis in our study. The results of SCT-3DI were best correlated with actual endoscopy findings and those found in the surgery. Based on the SCT-3DI, five patients were treated by open surgical techniques including laryngotracheal reconstruction procedures and dilation methods with hyoid grafts or stents. Two undertaken endoscopic treatment with CO2 laser excision of scar tissue or granulomas, one accepted no surgical procedures. All eight patients were successfully decannulated with improved voice and satisfied airway after 6 month postoperative follow-up. CONCLUSIONS: Evaluation of the upper airway with SCT-3DI is useful in the diagnosis, measurement, and definition of the laryngotracheal stenosis. Objective data obtained from SCT-3DI is very helpful for operative scheme. Patients treated with appropriate procedures best suited for every stenosis have reached satisfactory clinical results. PMID- 12889119 TI - Antioxidation properties and mechanism of action of dihydromyricetin from Ampelopsis grossedentata. AB - AIM: To assess the antioxidative properties and the mechanism of action of dihydromyricetin (DMY) from Ampelopsis grossedentata. METHODS: The antioxidative properties of DMY were measured by scavenging 1,1-diphenyl-2-picrylhydrazyl radical (DPPH) and inhibiting lipid peroxidation induced by FeSO4-edetic acid in linoleic acid. The mechanism of antioxidative properties of DMY was tested by measuring the chelating activities of DMY for Fe2+ with ultraviolet spectrum (UV) method. RESULTS: The specific absorption of DPPH radical solution at 517 nm was reduced 73.3%-91.5% when DMY was added into the reaction solution in the concentration range from 0.01% to 0.04%. DMY was shown to greatly inhibit the increase of lipid peroxidation (LPO) values in linolei acid system catalyzed by FeSO4-edetic acid. The reaction rates (A532.min-1) of lipid peroxidation were 0.0021-0.0004 in the concentration range from 0.01% to 0.04% and the inhibition activities of DMY was found to be in a concentration-dependent manner. The mechanism of antioxidative properties of DMY was chelating Fe2+ in the Fe(2+) dependent lipid peroxidation system. CONCLUSION: DMY showed great antioxidative effect and would be a good natural antioxidant. PMID- 12889120 TI - [Effect of lipopolysaccharide on expression of matrix metalloproteinase-9 in human synoviocyte from patients with rheumatoid arthritis]. AB - AIM: To study the effects of lipopolysaccharide (LPS), the supernatant of U937 cells stimulated with LPS and dexamethasone on matrix metalloproteinase-9 (MMP-9) expression in the synoviocyte from patients with rheumatoid arthritis(RA). METHODS: Fibroblast-like cells (FLS) from the joint tissue of patients with rheumatoid arthritis were cultured and incubated for 24 h with LPS (1 mg.L-1) or the supernatant of U937 cells stimulated with LPS (1 mg.L-1) for 24 h. Dexamethasone was added to the supernatant of U937 cells and FLS was incubated for 24 h. The activity of MMP-9 was analyzed by gelatin zymography. Protein expression of MMP-9 was detected by Western blot using special polyclonal antibodies. The mRNA expression of MMP-9 was detected by RT-PCR. RESULTS: The expression of MMP-9 was not markedly changed in FLS treated with LPS. The MMP-9 activity, MMP-9 secretion and MMP-9 mRNA expression were significantly increased in FLS cultured with the supernatant from U937 cell treated with LPS. Dexamethasone markedly inhibited the activity, protein secretion and mRNA expression of MMP-9 in FLS cultured with the supernatant from U937 cell stimulated with LPS, and the inhibitory effects were increased as the concentration of dexamethasone increased. CONCLUSION: LPS did not directly affect the expression of MMP-9 in FLS, but it was found to indirectly cause the increase of MMP-9 expression in FLS by stimulating U937 cell. Dexamethasone was found to inhibit this increase of MMP-9 expression. PMID- 12889121 TI - [Potentiation and mechanism of cisplatin-induced apoptosis by lidamycin in human hepatoma BEL-7402 cells]. AB - AIM: To investigate the synergetic effect and the mechanism of antitumor action of the antibiotic lidamycin in combination with cisplatin in vitro. METHODS: Cytotoxicity of the drugs was measured by clonogenic assay. Chromatin condensation was observed by co-staining with fluorescent dyes, Hoechst 33342 and propidium iodide. Apoptotic sub-G1 was detected by flow cytometry and DNA ladder was observed using agarose gel electrophoresis. Bcl-2 protein level was detected by Western blot assay. RESULTS: By using clonogenic assay, lidamycin in combination with cisplatin was found to have synergetic effects on the proliferation of human hepatoma BEL-7402 cells. The data showed that BEL-7402 cells treated with cisplatin and lidamycin in combination produced internucleosomal DNA fragmentation analysed by agarose gel electrophoresis. The results of flow cytometry showed that cisplatin and lidamycin administrated in combination showed no obvious change in G1 phase distribution compared with single treatment. However, this combination reduced the S phase arrest and reversed the reduction of G2/M phase induced by single treatment. The results also showed that there was 11.3% or 9.37% of cells undergoing apoptosis in BEL 7402 cells treated with cisplatin or lidamycin, respectively, while it showed 32.4% of apoptotic cells in combination treatment. Cisplatin, lidamycin and combination of cisplatin and lidamycin was shown to induce typical chromatin condensation in BEL-7402 cells. The study showed that 0.5 mumol.L-1 cisplatin or 1 x 10(-4) mumol.L-1 lidamycin alone decreased Bcl-2 protein level, while lidamycin in combination with cisplatin strongly inhibited expression of Bcl-2 proteins in BEL-7402 cells. CONCLUSION: The results suggest that lidamycin enhancement of cisplatin-induced apoptosis associates with decrease of Bcl-2 protein expression, which may be useful for cancer chemotherapy. PMID- 12889122 TI - Studies on antidiabetic effects of cortex Moutan polysaccharide-2b in type 2 diabetes mellitus rats. AB - AIM: To study the antidiabetic effects of cortex Moutan polysaccharide-2b (CMP 2b) in type 2 diabetes mellitus (T2DM) rats. METHODS: The T2DM model rats were induced by a single intravenous injection of low dose streptozotocin (STZ) and intake of high sucrose-fat diet. CMP-2b was given to T2DM rats daily through gavage for 4-5 weeks. The body weight, water and food intake, fasting blood glucose (FBG), glucose tolerance, plasma lipids, serum insulin, and insulin receptor (Ins R) were determined. RESULTS: Oral administration of CMP-2b significantly decreased water and food intake, FBG, total cholesterol (Tch), and triglyceride (TG), improved the impaired glucose tolerance (IGT), and remarkably raised the number of low affinity InsR and insulin sensitivity index (ISI) in T2DM rats. CONCLUSION: CMP-2b may be useful for treating T2DM and its complications. PMID- 12889123 TI - [Synthesis and antibacterial activity of pyridonecarboxylic acid derivatives containing 2-methyl-5-nitroimidazol]. AB - AIM: To study the synthesis and antibacterial activity of pyridonecarboxylic acid derivatives containing 2-methyl-5-nitroimidazol. METHODS: Pyridonecarboxylic acid derivatives containing 2-methyl-5-nitroimidazol were synthesized primarily from 2 methyl-5-nitroimidazol, norfloxacin, ciprofloxacin, enoxacin via nucleophilic substitution and esterification. The antibacterial activity of the nine target compounds were tested. RESULTS: Nine new compounds were synthesized (IIa-c and IIIa-f). The structure of the title compounds were identified by 1HNMR, MS as well as elementary analysis. CONCLUSION: Compounds IIa, IIb and IIc showed antibacterial activity, and were worth further studying. PMID- 12889124 TI - [Synthesis and antibacterial activity of 7-(4-acylamino-thiocarbamoyl-1 piperazinyl) fluoroquinolone analogues in vitro]. AB - AIM: To synthesize new fluoroquinolone analogues as antibacterial compounds. METHODS AND RESULTS: By reaction of acryl chloride(chloro-carbonic ester) with sodium sulfocyanate, acyl isosulfocyanic ester were easily obtained. Twelve 7-(4 acylamino-thiocarbamoyl-1-piperazinyl) fluoroquinolone analogues (1-12) were synthesized through modifying the 7-piperazine of norflorxacin and ciprofloxacin with isosulfocyanic ester synthesized above. The structures of synthesized compounds were characterized by 1HNMR, IR and elemental analysis. CONCLUSION: Antibacterial activities of the new compounds were evaluated in vitro compared with norflorxacin. Compounds 5, 7, 10 and 12 showed antibacterial activities. PMID- 12889125 TI - [Two new phenylethanoid glycosides from Corallodiscus flabellata]. AB - AIM: To study the chemical constituents from Corallodiscus flabellata. METHODS: Fresh plant of Corallodiscus flabellata was extracted twice with boiling water, filtered to remove insoluble materials, concentrated under reduced pressure at temperature 55 degrees C to a small volume. The concentrated liquor was subjected to solvent-solvent partitioning using ether, ethyl acetate, and n-butanol (saturated with water). The fraction of ethyl acetate extract was chromatographed over macroporous adsorption resin (Diaion HP-20) eluted with a mixture of H2O and MeOH in increasing MeOH content. Their fractions from resin were repeatedly chromatographed over Sephadex LH-20, Toyopearl HW-40, gel MCI, Gel CHP-20 and silica gel column. Structures of compounds obtained were identified on the basis of their spectral data, hydrolysis and chemical correlation. RESULTS: Two phenylethanoid glycosides (I, II) and three phenolic acids were obtained from the EtOAc fraction of water-extracts. Their structures were identified as 3,4 dihydroxyphenylethanol-8-O-[beta-D-apiofuranosyl (1-->2)]-beta-D-glucopyranoside (I), 3,4-dihydroxyphenylethanol-8-O-[(5-O- Vanilloyl)-beta-D-apiofuranosyl(1- >2)]-beta-D-glucopyranoside (II), vanillic acid (III), syringic acid (IV) and ferulic acid (V). CONCLUSION: I and II are new compounds. Compounds III, IV and V were isolated from this plant for the first time. PMID- 12889126 TI - [Studies on the chemical constituents of the aerial part of Cimicifuga foetida L]. AB - AIM: To look for new active constituents from the aerial part of Cimicifuga foetida L. METHODS: Various column chromatographic techniques were used for the isolation and purification of the principles. The structures were elucidated on the basis of spectral data and chemical evidences. RESULTS: Five 9,19-cycloartane triterpenoid saponins and one sitosterol saponin were obtained and identified as cimifoetiside I [12 beta-hydroxycimigenol-3-O-beta-D-galactoyranoside, (1)], cimifoetiside II [(23R,24R) cimigenol-3-O-beta-D-galactopyranoside, (2)], cimigenol-3-O-beta-D-galactopyranoside (3), 12 beta-hydroxycimigenol-3-O-beta-D xylopyranoside (4), 12 beta-hydroxycimigenol-3-O-alpha-L-arabinopyranoside (5), daucosterol (6). CONCLUSION: Compounds 1 and 2 are new and compounds 4 and 5 were isolated from this plant for the first time. PMID- 12889127 TI - [Alkaloids from a Tibetan medicine Meconopsis quintuplinervia Regel]. AB - AIM: To reinvestigate the chemical constituents of the ethanolic extract of Meconopsis quintuplinervia Regel which is a traditional Tibetan medicine used for treatments of hepatitis, tuberculosis etc.. METHODS: The compounds were enriched by column chromatography techniques over silica gel, macro porous resin and Sephadex LH-20 absorbents, and finally purified by reverse phase preparative HPLC methods with isocratic mobile phase systems of methanol-H2O-acetic acid (500:500:1) and acetonitrile-H2O-acetic acid (200:800:1). Structural determination of the pure compounds were based on extensive analyses of modern spectroscopic methods including IR, MS, HRMS, 1D- and 2D-NMR spectra. RESULTS: Three alkaloids were obtained and their structures were elucidated as norsanguinarine (I), O-methylflavinantine (II) and 6-methoxy-17-methyl-2, 3 [methylenebis (oxy)]-morphin-5-en-7-one (III). CONCLUSION: Norsanguinarine (I) was isolated from genus Meconopsis for the first time, and 6-methoxy-17-methyl 2,3-[methylenebis(oxy)]-morphin-5-en-7-one (III) is a new alkaloid named as meconoquintupline. PMID- 12889128 TI - [Determination of trigonelline by HPLC and study on its pharmacokinetics]. AB - AIM: To develop a sensitive and specific HPLC method for determination of trigonelline in rabbit plasma, and study the pharmacokinetics in rabbit. METHODS: After ig of fenugreek extract and i.v. of trigonelline in rabbit, the biological samples could be well purified after precipitation of protein with methanol and acetonitrile. Asahipak NH2P-50 column was used, the mobile phase consisted of acetonitrile-water (90:10) at a flow-rate of 1.2 mL.min-1, and detection wavelength was set at UV 265 nm. The column temperature is 30 degrees C. RESULTS: The calibration curve was linear in the range from 0.98 mg.L-1 to 31.28 mg.L-1, with r = 0.9986, the detection limit of this method was 50 micrograms.L-1. The concentration-time curves of trigonelline in rabbits after ig and i.v. administration were shown to fit one-compartment and two-compartment open model, respectively. The main parameters after ig of fenugreek extract were as follow: T1/2(Ka) was 0.9 h, T1/2(Ke) was 2.2 h, V was 0.64 L.kg-1, AUC was 1.93 mg.min.L 1. The main parameters after i.v. of trigonelline were as follows: T1/2 alpha was 10.8 min, T1/2 beta was 44.0 min, K21 was 0.044 min-1, K10 was 0.026 min-1, K12 was 0.017 min-1, AUC was 931.0 mg.min.L-1. CONCLUSION: Trigonelline showed a middle rate of absorption and fast rate of elimination in rabbit. Meanwhile, the method is simple, accurate, with a good reproducibility, and it provide a basic method for the investigation of trigonelline and fenugreek pharmacokinetics. PMID- 12889129 TI - [Determination of adenosine, rutin and quercetin in Carthamus tinctorius by HPCE]. AB - AIM: To develop a method for determination of adenosine, rutin and quercetin in Carthamus tinctorius L. by high performance capillary electrophoresis(HPCE). METHODS: A fused silica capillary (66.5 cm x 50 microns ID, an effective length of 58 cm) was used. The running buffer composed of 50 mmol.L-1 borax (pH 9.7) containing 18% methanol. The applied voltage was 24 kV and the capillary temperature was 20 degrees C. The detection wavelength was 210 nm. Rifampicin was used as internal standard. RESULTS: A good linearity between peak area ratio of the common peak to the internal standard and the concentration was found in the range of 10-160 mg.L-1 for adenosine, 100-2,000 mg.L-1 for rutin and 100-1,600 mg.L-1 for quercetin (r > 0.998). The average recoveries were 98.5%-100.5%, 96.9% 99.5% and 99.1%-99.5% for adenosine, rutin and quercetin, respectively. The relative standard deviation was less than 6.5% (n = 5). CONCLUSION: The method is simple, rapid and with satisfactory recoveries and good reproducibilities. It can be used to control the quality of Carthamus tinctorius. PMID- 12889130 TI - [Determination of chlorogenic acid and eriodictyol-7-O-beta-D-glucuronide in Pyrrosia by RP-HPLC]. AB - AIM: To develop a method for determination of chlorogenic acid and eriodictyol-7 O-beta-D-glucuronide in Pyrrosia of different species and different places of origin by RP-HPLC. METHODS: Chromatography was performed using a C18 column with mobile phase of methanol-water-phosphoric acid (50:200:0.2). The monitoring wavelength was 284 nm. RESULTS: The linear ranges were 0.01-5.0 micrograms (r = 0.9997) and 0.004-5.0 micrograms (r = 0.9997), the recoveries were 97.1% (n = 8, RSD = 2.7%) and 98.8% (n = 9, RSD = 2.5%) for chlorogenic acid and eriodictyol-7 O-beta-D-glucuronide, respectively. CONCLUSION: The method was employed to the analysis of 21 samples of Pyrrosia. The contents of compounds vary greatly depending on the species used, place of collection and time of harvesting. The HPLC method is sensitive, rapid and can be used to control the quality of Pyrrosia and to guide reasonable season of harvesting. PMID- 12889131 TI - [Study on Aconitum diterpenoid alkaloids from flowers of Aconitum kusnezoffi and its decoction by ESI-MS]. AB - AIM: To study the change of chemical compound after decocting the flowers of Aconitum kusnzoffii Reichb (FAK) by analyzing the alkaloids in FAK and the decoction of FAK qualitatively. METHODS: The alkaloid extracts of FAK and alkaloid mixtures in the decoction of FAK were directly analyzed by electrospray ionization trap tandem mass spectrometry via by pump. RESULTS: Three novel aconitum alkaloids were found in FAK. In addition, it is found that both diester diterpenoid alkaloids and triester diterpenoid alkaloids were hydrolyzed in the decoction of FAK. The hydrolysis products of the diester-alkaloids are benzoylaconines and aconines; the hydrolysis products of the triester-alkaloids are 3-acetyl-aconines. CONCLUSION: ESI-MS" method is suitable for the analysis of Aconitum alkaloids in the decoction of FAK as well as other Aconitum plants, which offers the advantages of convenience, speed, sensitivity and specificity. PMID- 12889132 TI - [Continuous in situ monitoring of the dissolution rate of solid pharmaceutical preparations using a multiple channel fiber-optic chemical sensor]. AB - AIM: To study the dissolution rate of solid pharmaceutical preparation on-line, a multiple channel fiber-optic chemical sensor based on fluorescence multiple quenching (FOCSMQ) without filtering and sampling was made. METHODS: Using the multiple channel FOCSMQ linked with computer, the dissolution rates of ofloxacin tablets, metronidazole tablets and nitrofurantoin tablets were monitored continuously on-line. The instrument can give the sample data, display the real time curve and calculate the T1/2 and td automatically. A computer was used to select the best function from five common fitting models to fit the dissolution curve. RESULTS: The average recoveries of the FOCSMQ method were 97.4%-104.4%, 97.4%-103.8% and 96.6%-102.1%. The RSDs (n = 6) of within-day and between-day were less than 5%. The parameters of the dissolution and all results of measurement using the instrument have no significant difference compared with the Chinese Pharmacopoeia (ChP) (2000) method and the United States Pharmacopoeia (USP) (23) method (P > 0.05). It does not need sampling and dilution, and never contaminate sample. It can shorten time of the experiment. CONCLUSION: The method is simple, rapid and reliable. PMID- 12889133 TI - [Study on polymethacrylate nanoparticles as delivery system of antisense oligodeoxynucleotides]. AB - AIM: To investigate the possibility of polymethacrylate nanoparticles (NP) for antisense oligodeoxynucleotides delivery system. METHODS: The nanoparticles were prepared by evaporating ethenol solution containing Eudragit RL100 or RS100, and then mixtured with oligonucleotides. The morphology and size were investigated by a transmission electron microscope and Mastersizer particle characterization systems, and the cytotoxicity was evaluated by Trypan Blue staining and hemolysis test. The flow cytometer was used to determine the uptake of fluorescence labelled oligodeoxynucleotides. RESULTS: The morphology of nanoparticles showed spherical and orderly, the average diameter was about 127 nm, and almost the antisense oligodeoxynucleotides (ODN) were loaded when NP: ODN was 6.6. The uptake of ODN was significantly increased when loaded by nanoparticles, which well depended on the nanoparticles concentration. Meanwhile, slightly cytotoxicity was observed when high dose of nanoparticles was used. CONCLUSION: The polymethacrylate nanoparticles appeared to be a promising vehicle for gene delivery. PMID- 12889134 TI - [Preparation of solid lipid nanoparticles by solvent diffusion method]. AB - AIM: To establish an efficient method for preparation of solid lipid nanoparticles with high recovery. METHODS: Monostearin solid lipid nanoparticles was prepared by solvent diffusion method in aqueous system. The recovery of the method was greatly improved by adjusting the Zeta potential. RESULTS: The drug loaded solid lipid nanoparticles suspension was quickly produced and easily separated with centrifugation at 4,000 r.min-1 under acidic condition. Compared with the nanoparticles made without adjusting the Zeta potential, the recovery of nanoparticles prepared in this way was significantly increased. The release behavior in vitro showed an initial burst effect in the first 3 hours followed by a slower rate stage of 4 days with nearly 6% drug released in each day. CONCLUSION: The solvent diffusion method in aqueous system might be used as a new method to prepare solid lipid nanoparticles in the future. The loaded drug can be released in a controlled manner. PMID- 12889135 TI - [Two chemotypes of Pogostemon cablin and influence of region of cultivation and harvesting time on volatile oil composition]. AB - AIM: To analyze and compare the constituents of the volatile oil of Pogostemon cablin collected from different regions of cultivation and harvesting times in order to evaluate the quality of Shipai Huoxiang and to expound the chemical intension of Pogostemon cablin. METHODS: The combination of GC and MS. RESULTS: The volatile oil compositions of Herba Pogostemonis collected from various of cultivation regions and harvesting times are obviously different. Based on the chemical differences of the volatile oil compositions, Pogostemon cablin is divided into two chemotypes, Pogostone-type and Patchouliol-type. The former was cultivated in Guangzhou and Gaoyao regions, locally named as "Shipai Huoxiang"; the latter was locally named as "Hainan Huoxiang", cultivated in Wuchuan, Suixi and Leizhou regions of Guangdong Province and Wanning region of Hainan Province. The Pogostone-type contains rich oxygenated components, especially pogostone in the volatile oil compositions and poor non-oxygenated composition with patchouliol. The above chemical data may be used as evaluation standard for the authentic Shipai Huoxiang. The Patchouliol-type contains similar quantities of oxygenated and non-oxygenated composition, especially rich patchouliol with poor pogostone in oxygenated compositions, rich delta-guaiene and alpha-guaiene in non oxygenation compositions. The contents of volatile oil and their constituents were varied due to different harvesting time. The yields of pogostone and volatile oil of Shipai Huoxiang was higher in July. The quality of the samples collected in this month was better. CONCLUSION: According to the volatile oil compositions, there are two chemotypes (Pogostone-type and Patchouliol-type) in Pogostemon cablin plant. These two chemotypes correspond to the genotypes of plastid matK gene and nuclear 18s rRNA gene by cluster analysis. PMID- 12889136 TI - [Biological characteristics of microtubule and related drug research]. PMID- 12889137 TI - [Application of cationic polymer vector for gene delivery systems]. PMID- 12889138 TI - [Reminiscences of the trail of my research]. PMID- 12889139 TI - [Central dogma of molecular biology]. PMID- 12889140 TI - [Ion channels on T lymphocyte]. AB - There is increasing evidence to show that ion channels on lymphocytes play a very important role in the regulation of immune functions. In T lymphocytes, there are three types of ion channels on cell membrane: Ca2+, K+ and Cl- channel. The influx of Ca2+ into T lymphocyte through Ca2+ channel (CRAC) may act as a second messenger to activate T lymphocyte when antigen binds to the receptor (TCR). The efflux of K+ from T lymphocyte through the K+ channel contributes to the formation of T cell membrane potential. The level of the membrane potential may affect the influx of Ca2+ into T cells. Therefore, the activation and the functions of T cell can be regulated by K+ channel indirectly. Cl- channel in T lymphocyte was found in recent years and it is probably involved in the regulation of cell volume. The recent progress on ion channels in T lymphocyte is summarized briefly in the present paper. PMID- 12889141 TI - [Neurogranin: a brain-specific protein]. AB - Neurogranin (Ng) is a newly discovered brain-specific protein composed of 78 amino acid residues, which mainly located postsynaptically in the cerebral cortex, hippocampus and olfactory bulb in adult human or animals. As a member of calpacitin family, Ng is a protein kinase C (PKC) substrate and calmodulin (CaM) reservoir. In the physiological conditions, Ng forms a complex with CaM, and its CaM-binding affinity was modulated by phosphorylation, oxidation and glutathiolation under the activation of PKC or oxidant stress, which may be involved in the regulation of CaM and CaM-activated proteins, such as CaM dependent nitric oxide synthase (NOS), CaM-dependent protein kinase II (CaMKII) and CaM-dependent adenylate cyclase (AC). Since most of CaM-activated proteins were involved in long-term potentiation (LTP) and long-term depression (LTD), and the timing pattern of Ng gene expression and protein synthesis are coincidence with synaptogenesis and development, it is suggested that Ng may play an important role in learning, memory and neuroplasticity. In addition, it was found that the changes of Ng expression might associate with certain cerebral pathophysiologic disorders, such as hypothyroidism, sleep-deprivation, brain aging and cerebral hypoxic preconditioning. PMID- 12889142 TI - [New targets for drug therapeutics: receptors for purines and pyrimidines]. AB - Endogenous nucleoside and nucleotide mediate a lot of functions via cell surface P receptors (receptors for purines and pyrimidines) in many organs. Nucleoside and nucleotide have protective roles in the events such as cancer, apoptosis, ischemia, wound healing, osteoporosis, drug toxicity, inflammation and pain. Moreover, the development of selective agonists and antagonists for P1, P2 receptors and P receptor subtypes may provide novel drugs in therapeutic strategies. PMID- 12889143 TI - [Advances in the applications of DNA chips and proteomics approaches for neurosciences]. AB - DNA chips and proteomics are two of the recently developed high-throughput technologies that allow us to simultaneously analyze the expression levels of multiple genes and the interactions of their products in the brain. Their applications in neuroscience provide us with the unprecedented opportunities for understanding of the brain. A typical gene chip experiment contains a series of steps, including preparations (or purchase) of DNA chips, target DNA and probes, hybridization of chips with target DNA, chip scanning, and imaging analysis. The technologies in proteomics are more complicated, which comprise of three main aspects of technologies: protein isolation, identification and bioinformatic analysis of data. If protein isolation is based on 2-D, it includes the preparation of protein sample, 2-D, staining gels, spot excision, proteolysis, mass spectrometry of purified protein and finally undergoing bioinformatic analysis. Here we review the two technologies with emphasis on their applications in neurosciences. The challenges, advantages and disadvantages of the two techniques, and perspectives for their developments are discussed. PMID- 12889144 TI - [The mechanisms of a new plant lectin FRIL from dolichos lablab that preserves quiescent hematopoietic stem cells in suspension culture]. AB - Lectins, proteins that bind sugar moieties of glycolipids and glycoproteins, occur in all organisms and are especially abundant in many plants. Plant lectins probably have multiple roles related to their ability to act as recognition molecules. Here we present a review on the biological characterization of a new plant lectin in red kidney bean named Flt3 receptor-interacting lectin(FRIL), and focus on the mechanisms that FRIL can preserve quiescent hematopoietic stem cells in suspension culture. PMID- 12889145 TI - [Gene families of G protein subunits]. AB - G proteins are heterotrimers composed of alpha, beta, and gamma subunits. So far 16 alpha, 6 beta, and 12 gamma subunit genes have been described in mammals. G protein subunit gene families are comparatively conservative and primitive, with all of which exon-intron junctions conform to the GT-AG rule except for one alternative splice site at 5'-end of exon 4 in Gnas which uses GT-TG junction. In the coding regions, exon-intron structures and intron positions among each of the subunit gene families are comparatively conserved. Most of the G protein genes have some characteristics of housekeeping genes. Analysis of distribution of G protein subunit genes in mammalian genomes reveals a tendency of clustering, with five pairs of the alpha subunit genes arranging as tandem duos. PMID- 12889146 TI - [Progress in the study of muscle satellite cells]. PMID- 12889147 TI - [Piwi gene family: the important gene modulating the divisions of stem cell]. PMID- 12889148 TI - [Research progress in the study of neural stem cell derived from spinal cord]. PMID- 12889149 TI - [Physiopathologic mechanisms of synovium with rheumatoid arthritis]. PMID- 12889150 TI - [Digital body]. PMID- 12889151 TI - [The regulation of metallothionein expression and its relationship to zinc]. PMID- 12889152 TI - [Signaling induced by HCO3- in sperm capacitation]. PMID- 12889153 TI - [Muscarinic receptor m5 subtype: an emerging orphan receptor]. PMID- 12889154 TI - [Progress of glial growth factor]. PMID- 12889155 TI - [A review of hippocampal place cell]. PMID- 12889156 TI - [The application of endothelial progenitor cells (EPC) in the cellular and gene therapy of cardiovascular-disease]. PMID- 12889157 TI - [Hypoxia-inducible factor 1 and mechanisms of hypoxia signal transduction]. PMID- 12889158 TI - [Progress in the studies on the CYP2C19 polymorphism]. PMID- 12889159 TI - [A study of nucleolar stress/injury and heat shock proteins]. PMID- 12889160 TI - [Application of gene manipulation in research of CYP450s]. PMID- 12889161 TI - [Progress in the study on the new member TALL-1 of TNF family]. PMID- 12889162 TI - [Gleevec: an novel inhibitor of bcr-abl tyrosine kinase]. PMID- 12889163 TI - [Recent research of the LIM protein family]. PMID- 12889164 TI - [Secretases as targets for the treatment of Alzheimer's disease]. PMID- 12889165 TI - Imaging of Alzheimer's disease. AB - Imaging is widely used in the evaluation of Alzheimer's disease (AD). In addition to imaging's traditional role of aiding in the exclusion of diseases that may be confused with AD, structural and functional imaging are being explored for potential use in the early detection of AD and as surrogate markers of treatment outcome. Volumetric measurements of the hippocampal and entorhinal regions using modern magnetic resonance imaging (MRI) and computed tomography are most widely studied in the discrimination of AD from nondemented aging and other dementias. Other imaging modalities include positron emission tomography and single-photon emission computed tomography, which evaluate global and regional disturbances of blood flow and metabolism, and proton spectroscopy, which assesses neuronal degeneration and loss as marked by the presence of abnormal metabolites. Diffusion tensor and functional MRI, which examine white matter changes and brain behavior relationships, are more recent techniques that contribute to the burgeoning application of imaging techniques to dementing illnesses and are notably improving our understanding of the pathophysiology of AD. PMID- 12889166 TI - Neuroimaging curriculum for neurology trainees: report from the Neuroimaging Section of the AAN. AB - Neuroimaging plays a major role in the evaluation of patients with neurological disorders. Surveys of neurologists have revealed that most rely on their own readings of images for patient management, and a majority believe that neurologists should be allowed to officially interpret and bill for scan reviews. The importance of neuroimaging training for neurology residents has been stressed by the Association of University Professors of Neurology. Although there is a desire to promote the neuroimaging education of neurologists, no curricula have existed previously. The Neuroimaging Section of the American Academy of Neurology (AAN) developed a task force of practicing neuroimagers to provide a neuroimaging curriculum for neurological trainees and training directors. The resulting curriculum is available on the Web sites of the AAN (http://www.aan.com) and the American Society of Neuroimaging (http://www.asnweb.org/education/curriculum.shtml) and will be updated as the need arises through evolving technology or breadth of applications. This curriculum should help in the design of neurology residency and fellowship programs and subspecialty pathways in which adequate neuroimaging education and training are desired for various reasons, including certification and the demonstration of competency and proficiency. PMID- 12889167 TI - Paradoxical emboli from calf and pelvic veins in cryptogenic stroke. AB - PURPOSE: The increased prevalence of patent foramen ovale in patients with cryptogenic strokes suggests the occurrence of paradoxical embolism. The identification of deep venous thromboses (DVTs) in this population would strengthen this hypothesis. The purpose of this study was to image the subdiaphragmatic venous system in a cohort of patients with cryptogenic strokes. MATERIALS AND METHODS: In 37 patients with cryptogenic brain ischemia and interatrial communication, duplex studies of calf, popliteal, and femoral veins, and magnetic resonance imaging venograms of the pelvis veins were performed. RESULTS: In 10 patients, DVTs were diagnosed that were considered to be the cause of cryptogenic brain ischemia on probable (n = 6) or possible (n = 4) bases. In these patients, the median time from stroke to DVT was 3.25 days. In 5 of these 10 patients, DVTs did not involve popliteal and femoral veins, areas thought most important to pulmonary embolism, but instead were isolated to calf or pelvic veins. Although none of these 10 patients had abnormal blood hypercoagulation tests, 8 of the 10 did have clinical conditions suggesting predisposition to developing DVTs, such as concomitant neoplasms or pulmonary embolism. CONCLUSIONS: Increased evidence for paradoxical embolism may emerge when diagnostic strategies use multiple imaging methods and evaluate a broad extent of the subdiaphragmatic veins. PMID- 12889168 TI - The prevalence of asymptomatic intracranial large-vessel occlusive disease: the role of diabetes. AB - OBJECTIVE: To investigate the prevalence of asymptomatic intracranial stenosis using transcranial Doppler ultrasonography in patients with evidence of asymptomatic carotid stenosis. BACKGROUND: Symptomatic atherosclerotic intracranial largeartery stenosis accounts for approximately 10% of ischemic strokes annually. It is unknown whether a significant risk for stroke is associated with asymptomatic intracranial stenosis, especially in patients with known asymptomatic carotid disease. DESIGN AND METHODS: Transcutaneous real-time B-mode Doppler ultrasonography was performed on 510 patients referred for the evaluation of asymptomatic carotid bruits. A peak systolic flow velocity > 1.40 m/s indicated carotid stenosis. Transcranial Doppler ultrasound was performed to identify intracranial large-artery disease. The peak systolic flow velocity indicating stenosis was > 120 cm/s for anterior circulation and > 100 cm/s for posterior circulation. Demographic and cerebrovascular risk factor information was recorded at the time of examination. Chi-square analysis with Pearson correction was performed to examine the significance of the findings. RESULTS: Five hundred ten patients (252 male, 258 female) aged 71.4 +/- 10.45 years were studied. The laboratory's accuracy was previously established as 93% for each technique for laboratory certification. Two hundred patients (39.2%) were found to have extracranial carotid stenosis, and 66 (12.9%) were found to have intracranial stenosis. Thirty-seven patients (56%, P < .01) were found to have concurrent stenosis. In patients with intracranial stenosis, 19 (28.8%, P < .03) had diabetes, and 26 (39.4%, p < .03) had coronary disease. There was no corresponding correlation with extracranial carotid disease and diabetes or coronary artery disease. CONCLUSIONS: The prevalence of patients with asymptomatic intracranial stenosis with concurrent carotid stenosis was greater than expected. Coexisting diabetes and coronary disease were significant risk factors in this population. Doppler ultrasonography may be a useful technique in delineating asymptomatic intracranial stenosis in those patients with existing carotid stenosis. Further studies are needed to clarify the risk of this population for stroke from asymptomatic intracranial stenosis and to determine optimal therapy. PMID- 12889169 TI - Quantification of hippocampal signal intensity in patients with mesial temporal lobe epilepsy. AB - BACKGROUND: Refractory mesial temporal lobe epilepsy (MTLE) is frequently associated with hippocampal atrophy (HA) and an abnormal hippocampal signal (Hsig) on magnetic resonance imaging (MRI). OBJECTIVE: To quantify Hsig on MRI using a simplified technique. METHODS: The authors included 15 patients with refractory MTLE who underwent surgery and had preoperative MRI with hippocampal volumetry. Hsig was quantified on preoperative coronal T1-weighted and T2 weighted MRI using the NIH-Image program. Hsig was determined for the head, tail, and entire hippocampal extension. Abnormal Hsig was defined when values were above (for T2) or below (for T1) 2 standard deviations from the mean of normal control group. RESULTS: The lateralization of abnormal Hsig values was concordant with electroencephalograms and HA in all patients. There was a significant difference in ipsilateral T2 Hsig between patients and controls (P < .0001), but not for contralateral T2 Hsig. T1 Hsig showed bilateral abnormalities more frequently, whereas T2 Hsig could lateralize better hippocampal abnormalities. Overall, there were no differences when comparing T2 Hsig for the entire hippocampus, head, or tail only. However, there were some individual differences. T2 Hsig abnormalities involved the entire hippocampus in most patients but were restricted to the head in one patient and to the tail in another. CONCLUSIONS: This simplified method for the quantification of Hsig using NHI-Image is an efficient method for the identification and quantification of hippocampal abnormalities in patients with MTLE. The assessment of the entire extension of the hippocampal formation may provide important additional data, compared to T2 relaxometry maps limited to one segment of the hippocampus. PMID- 12889170 TI - MRI brain T1 relaxation time changes in MS patients increase over time in both the white matter and the cortex. AB - OBJECTIVE: To test the sensitivity of whole-brain T1 relaxometry to the evolution of pathological changes in multiple sclerosis (MS). BACKGROUND: T1-weighted hypointense lesion load in the brains of patients with MS is associated with axonal loss. Other work has shown that T1 measurements may provide information complementary to existing imaging techniques, such as magnetization transfer imaging. METHODS: The authors studied 14 MS patients twice over a median time interval of 19.5 months (range, 14-22 months). Structural images and whole-brain T1 maps using a novel rapid-scanning technique (3 min/study) were performed at 3 T. Analysis focused on defining changes separately in the lesional and normal appearing white matter (NAWM) and in the cortical gray matter. RESULTS: At baseline, there was an inverse relationship between disease duration and the NAWM T1 histogram peak height (r = -0.75, P = .03). The total white matter T1 histogram peak height decreased over time (P < .001). This could be accounted for by changes in the NAWM (P < .03). There also was a decrease (6%) in the mean (11 of 14 patients, P = .004) and in the median (7%) (13 of 14 patients, P < .001) neocortical gray matter T1 over the follow-up period. CONCLUSIONS: Brain T1 maps can be generated quickly and are sensitive to pathological changes over time. T1 values in both the gray and the white matter at the baseline visit were related to disease duration, suggesting that the T1 changes are clinically relevant. Although the absolute values will be different, it is likely that similar changes will be able to be detected at 1.5 T. The role of T1 measurement as a magnetic resonance imaging outcome measure in clinical trials now should be explored. PMID- 12889171 TI - Brain CT and MRI findings in 100 consecutive patients with intracranial tuberculoma. AB - OBJECTIVE: The characteristics of intracranial tuberculoma on computed tomography (CT) and magnetic resonance imaging (MRI) are not well known. The authors reviewed the features of tuberculoma on CT scans and MRI. The authors also correlated the MRI characteristics on various pulse sequences with neuropathological findings. METHODS: The charts of patients with intracranial tuberculoma were consecutively reviewed during the period from 1988 to 1999. The diagnosis of tuberculoma was based on pathology (n = 19), clinical or neuroimaging response to tuberculous chemotherapy (n = 57), or evidence of systemic tuberculosis (n = 24). Neuropathological specimens of tuberculoma were graded for the amount of fibrosis, gliosis, necrosis, and cell types (lymphocytes, macrophages, plasma cells). RESULTS: One hundred patients (43 men) were identified. The age range was 1 to 75 years (mean = 30 y). The number of lesions ranged from 1 to > 100 (mean = 4.5 lesions/patient). Thirty-one patients had solitary lesions, whereas 69 had multiple lesions. Thirty-seven patients had hydrocephalus. After contrast administration, > 450 lesions were seen on CT and MRI images in all 100 scans. The diameter of these enhancing lesions ranged from 1 mm to 5 cm. Lesions > 1 cm showed varied enhancement, including irregular shapes, ringlike shapes, open rings, and lobular patterns. Targetlike lesions were seen in only 2 patients. Other features included cortical and subcortical infarcts (12 patients), calcification (10% of lesions), edema (33 patients), meningeal enhancement (12 patients), mass effect, and/or midline shift (18 patients). A hypointense core with a hyperintense rim was the most common signal characteristic on T2-weighted MRI. The central hypointensity on T2-weighted and fluid-attenuated inversion recovery (FLAIR) images reflected extensive necrosis and hypercellularity. CONCLUSION: Multiple tuberculomas and infratentorial locations were more common in the authors' patient population than in previous reports. The MRI signal characteristics of intracranial tuberculoma are extremely diverse. An isointense or hypointense core with a hyperintense rim on T2-weighted and FLAIR images is the most common presentation. Core hypointensity of lesions on these images is related to necrosis and the large number of cells. PMID- 12889172 TI - Symmetry of cerebral hemodynamic indices derived from bilateral transcranial Doppler. AB - OBJECTIVE: The authors evaluated with bilateral transcranial Doppler (TCD) ultrasonography the norm of interhemispheric difference for several cerebrovascular hemodynamic parameters. METHODS: Forty-four volunteers (33 male, 11 female; average age = 21 years; range, 20-23 years) were studied. The authors recorded bilateral systolic, diastolic, and mean flow velocity (FV) and noninvasive systolic, diastolic, and mean arterial blood pressure. Calculated indices included Gosling's pulsatility index (GPI), an index of autoregulation (Mx), critical closing pressure (CCP), and a noninvasive estimator of cerebral perfusion pressure (nCPP). All indices were averaged per side and patient. For each parameter, the left-right correlation coefficient (r2) and the 95% confidence limit of the left-right differences were calculated. RESULTS: All TCD derived indices displayed significant correlations between the left and right sides: r2 = 0.49 for mean FV (FVm), r2 = 0.66 for GPI, r2 = 0.79 for Mx, r2 = 0.93 for CCP, and r2 = 0.94 for nCPP. The 95% confidence intervals for the left right differences were 20 cm/s for FVm, 0.16 for GPI, 0.18 for Mx, 13 mm Hg for CCP, and 4.6 mm Hg for nCPP. CONCLUSIONS: This study gives the reference values for the assessment of left-right symmetry of cerebral hemodynamics using various TCD-derived indices. These reference values should be useful for clinical studies assessing the left-right asymmetry of cerebral hemodynamics on a daily basis. PMID- 12889173 TI - Variability and validity of a simple visual rating scale in grading white matter changes on magnetic resonance imaging. AB - BACKGROUND AND PURPOSE: A new simple visual rating scale can be used in magnetic resonance imaging (MRI) to grade the severity of white matter changes (WMC). The authors sought to study the interobserver variability and the validity of this visual rating scale against a computer-aided quantitative method in measuring WMC. METHODS: The authors examined 220 magnetic resonance images from stroke-free patients with traditional risk factors for atherosclerosis and middle cerebral artery stenosis who were participants in a clinical trial requiring MRI examination. The severity of WMC was graded from 0 (no WMC) to 3 (severe WMC). For the quantitative assessment of WMC, the volume of WMC were measured with a computer-aided, automated segmentation method. The interobserver variability was also determined. RESULTS: The mean volumes were significantly different among the different groups, which were scored 1, 2, and 3 (1324.59, 6475.48, and 25,440.05 mm3, respectively; 1-way analysis of variance, P < .001). Interobserver variability for grading WMC was good in different locations of the brain. Excellent results were found in the frontal lobe (kappa = 0.829) and the parietal occipital lobe (kappa = 0.646), whereas poor results were found in the basal ganglia (kappa = 0.391). There was perfect agreement between the 2 observers in 89.1% for the frontal lobe and 77.0% for the parietal-occipital lobe. CONCLUSIONS: The results suggest that this simple visual rating scale is a valid and reliable method for grading WMC in the lobar region but not in the basal ganglion. PMID- 12889174 TI - Subcortical hypointensity in partial status epilepticus associated with nonketotic hyperglycemia. AB - Transient gray and white matter magnetic resonance signal changes have frequently been reported because of underlying epileptic activity. However, transient subcortical hypointensity on T2 and fluid-attenuated inversion recovery images in the setting of status epilepticus (SE) has rarely been reported. The authors note 3 of 6 retrospective cases with partial SE associated with nonketotic hyperglycemia showing transient subcortical T2 hypointensity near the epileptic focus. These changes may be due to the accumulation of free radicals and iron deposition related to a hyperglycemia-induced hypoxic-ischemic state. PMID- 12889175 TI - Magnetic resonance imaging of meningoradiculomyelitis in early disseminated Lyme disease. AB - Lyme disease, a multisystem illness caused by the spirochete Borrelia burgdorferi, is the most common vector-borne disease in the United States. There are 3 clinical stages of Lyme disease: early localized, early disseminated, and late persistent disease. Neuroborreliosis, infection of the nervous system by B. burgdorferi, may occur during early disseminated or late persistent disease. Spinal cord involvement in early disseminated disease is extremely rare. In patients with early disseminated neuroborreliosis, treatment with antibiotics often leads to rapid recovery and may prevent further complications of Lyme disease. The authors present the clinical and radiographic findings, both before and after treatment, in a patient with meningoradiculomyelitis due to early disseminated Lyme disease. PMID- 12889176 TI - Monitoring the effect of chemotherapy in a mixed glioma by C-11-methionine PET. AB - The effect of chemotherapy with procarbazine, CCNU, and vincristine in an anaplastic oligoastrocytoma was monitored by repeated positron emission tomography (PET) with C-11-methionine (C-11-MET). Chemotherapy caused a continuous decline of active tumor volume at a rate of approximately 2.4% per day, resulting in complete remission that persisted until the end of follow-up at 3 years. Thus, the authors conclude that C-11-MET PET may be useful for monitoring chemotherapy in gliomas and deserves further study. PMID- 12889177 TI - Importance of jugular valve incompetence in contrast transcranial Doppler ultrasonography for the diagnosis of patent foramen ovale. AB - Transcranial Doppler (TCD) ultrasound with the intravenous injection of agitated saline as contrast (cTCD) is an effective method for detecting right-to-left intracardiac and extracardiac shunt (RLS); however, the sensitivity of cTCD in the diagnosis of RLS remains slightly less than that of transesophageal echocardiography, even in patients with adequate transtemporal ultrasonic bone windows. The authors present a case with cTCD underestimating RLS because of jugular valve incompetence in a 42-year-old man presenting with an episode of transient aphasia. Three weeks after transcatheter closure of a patent foramen ovale associated with an atrial septal aneurysm, he experienced 2 episodes of amaurosis fugax. Following a negative 45-minute embolus detection study with power M-mode TCD, the patient underwent a cTCD study with monitoring of the left middle cerebral artery (MCA), the anterior cerebral artery, and the submandibular extracranial internal carotid artery. A single microbubble (MB) was detected in the left MCA in only 1 of 5 studies; the remaining runs all failed to detect an RLS. Significant MB reflux was noted in the left internal jugular vein because of jugular valve incompetence. The authors conclude that incompetence of the jugular vein valve can result in a false negative cTCD study for RLS detection. PMID- 12889178 TI - Pituitary apoplexy in the setting of coronary angiography. AB - Pituitary apoplexy (PA) is a rare but potentially life-threatening event. Typically, it results from hemorrhage into a pituitary adenoma, although cases affecting the intact gland have been reported. PA may occur spontaneously or in a setting of certain diagnostic and therapeutic procedures. The association of PA and contrast administration in the setting of neuroimaging have been postulated. The authors report a case of PA following coronary angiography. To the best of the authors' knowledge, this scenario has not been previously reported. A 66-year old woman with a congenital septal defect underwent coronary angiography that was technically uncomplicated. After the procedure, the patient experienced retro orbital pain and developed ophthalmoplegia. Magnetic resonance imaging (MRI) showed a hemorrhagic sellar mass extending into the cavernous sinus. A 3-month follow-up MRI revealed cystic changes. The patient expired 4 months later because of cardiorespiratory failure. PA in the absence of adenoma was confirmed on autopsy. The authors hypothesize that apoplexy here was related to the administration of Omnipaque in combination with the anticoagulation effect of heparin. PMID- 12889179 TI - Functional magnetic resonance imaging (fMRI) of a stutterer's brain during overt speech. PMID- 12889180 TI - [Leg re-contouring by using a new technique of partially removing gastrocnemius]. AB - OBJECTIVE: To evaluate a method to partially remove gastrocnemius for improvement of the contour of the leg. METHODS: Thirteen patients with bulked calf were undergoing the treatment. With a self-designed apparatus, the gastrocnemius was partially removed by placing the device in the muscle through an incision in the popliteal fossa. The follow-ups were carried out for 1-6 months. RESULTS: Thirteen patients were successfully treated by above mentioned technique. The removed amount of muscle was weighted between 11 g and 201 g, averaged 77 g. All of the patients could be able to walk three days after the operation. But, the normal walk had to take 1 month after of the surgery for recovering. The function of the ankle joint was not obviously influenced. CONCLUSION: The above mentioned technique is a safe and effective method for re-contouring the leg. PMID- 12889181 TI - [Penile reconstruction by using a scapular free flap]. AB - OBJECTIVE: To evaluate the scapular free flap for penile reconstruction. METHODS: Six patients with penile loss were reconstructed by transferring a scapular free flap with implanting a Malleable penile prosthesis. RESULTS: All of the flaps survived completely. With the follow-ups from six to twelve months, the reconstructed penile had excellent configuration and good function. CONCLUSION: The above mentioned technique could be a good method for penile reconstruction. PMID- 12889182 TI - [A comparative study of EGFR and FGFR-2 expression in fetal and adult skin]. AB - OBJECTIVE: To observe the expression characteristics of EGFR and FGFR-2 in normal skin from fetal and adult, and attempted to probe the molecule mechanism of fetal scarless healing. METHODS: The skin samples of fetal and adult were taken from abortive fetus of obstetrics unit and donor site of plastic operation patients in our burn unit, respectively. EGFR and FGFR-2 were used as the biochemical markers for reparative cells. Immunohistochemistry staining technique was employed to determine the expressive levels of different epithelial cells markers. RESULTS: There were EGFR and FGFR-2 antibody positive cells in normal skin from fetal and adult, but the expressive levels of EGFR and FGFR-2 protein had apparent difference, with the time of fetation increasing, the EGFR and FGFR-2 positive expression rate became stronger gradually. The number of FGFR-2 antibody positive cells found in adult skin was much more than that in fetal skin. CONCLUSION: There were the inherent differences of EGFR and FGFR-2 antibody immunohistochemistry staining in cells of adult and fetal skin. which may be an essential facet of fetal scarless healing. PMID- 12889183 TI - [Cell cycle analysis and P53 gene mutation detection of fibroblasts derived from the surrounding skin of keloids]. AB - OBJECTIVE: To investigate whether there are abnormal fibroblasts in the surrounding skin of keloids for better understanding of the etiological feature of keloids. METHODS: Fresh samples were used for cell culture. Flow cytometry was used for analyzing cell cycles of fibroblasts derived from keloids and the surrounding skin. Proliferative cell proportions were compared between every two groups. P53 exon 4, 5 and 6 were amplified by PCR. DNA was sequenced to examine the structure of the destination gene. RESULTS: The proliferative cell proportion of the fibroblasts derived from the surrounding skin is not so high as that from the verge of keloids, but is higher than the normal skin fibroblasts derived from other part of the keloid patients or persons without keloids (P < 0.05). Mutations (point and frameshift mutations) of P53 exon 4 (6/6) and exon 5(2/6) in fibroblasts derived from the surrounding skin of keloids were identified. Fibroblasts derived from keloids have the same mutations as its surrounding skin. CONCLUSION: There are abnormal fibroblasts in the surrounding skin of keloids. It may be the consequence of keloid infiltrative growth and a reason of easy recurrence of keloid after therapy. PMID- 12889184 TI - [Modulation of the neutrophil died process-mechanisms of the flap protection from ischemia-reperfusion with dexamethasone]. AB - OBJECTIVE: To study the modulation of neutrophil apoptosis and necrosis, elucidate the protecton mechanisms of dexamethasone. METHODS: 50 rats were divided into 5 groups. I: normal island flap. II: artery occlusion for 8 hours, saline given into peritoneal. III: venous occlusion for 8 hours, saline given into peritoneal. IV: artery occlusion for 8 hours, dexamethasone injected through peritoneal. V: venous occlusion for 8 hours, dexamethasone injected through peritoneal. Flap survived areas were measured, neutrophil apoptosis and necrosis values were ananlyzed with FCM. Their morphology were observed with light microscopy. The swallows of apoptotic neutrophil by mascrophages were studied with transmission electron microscopy. RESULTS: Flap survived areas I, IV and V groups were higher than that in II and III groups, which in I, IV, V had no significant values. Apoptotic neutrophils values in II and III groups on 1, 3 days were less than that in, but on 6 days, higher than that in I, IV and V. Necrotic neutrophils showed another tendency postoperatively when compared with its apoptotic values. There were more swallows of apoptotic neutrophils in flaps of IV, V groups than that in II, III flaps. CONCLUSION: Dexamethasone's flap protection results from modulating of neutrophils apoptosis, decreasing it's necrosis, increasing swallowings values of apoptotic neutrophils by macrophages. PMID- 12889185 TI - [Transferring neurovascular rectus femoris muscle segment for treatment of facial paralysis]. AB - OBJECTIVE: To investigate a new technique for functional treatment of chronic facial paralysis. METHODS: Based on anatomy of intramuscular neurovascular structure in the rectus femoris muscle, 7 consecutive patients with facial paralysis were treated by using a technique of microsurgically free-transferring neurovascular rectus femoris muscle segment to the face in one-stage. Follow-ups were 10 to 24 months. RESULTS: All of the 7 patients showed significantly improvement in the appearance of the oral commissure and oral competence. No complications occurred in the donor site. CONCLUSIONS: The above mentioned technique may have the advantages of preventing the intramuscular nerve and vessel from the surgical injury during splitting the muscle. It could also maintain the transferred muscular segment in a proper tension in the recipient site. PMID- 12889186 TI - [The transplantation of latissimus dorsi flap of the base of T shape artery with the pedicle of the thoracodorsal artery]. AB - OBJECTIVE: To explore the possibility that the free latissimus dorsi musculo cutaneous flap to repair the forearm leg wound. METHODS: To design latissimus dorsi musculo-cutaneous flap which is foundation on T form thoracodorsal artery stalk. To set the short arm into the receiver artery break and anastomos them. It is not only reassure the blood of free musculo-cutaneous flap, but also reconstruct the continuation of the receiver main artery. RESULTS: In 16 patients, 15 patients success completely, 1 patient main success. The blood supply of receiver is adequate. CONCLUSIONS: The free T form thoracodorsal artery stalk musculo-cutaneous flap free grafting is a good method to repair the skin and soft tissues defection of forearm and leg. PMID- 12889187 TI - [Repairing large alar defects by using a retro-grade auricular flap]. AB - OBJECTIVE: To evaluate a retro-grade auricular flap for repairing large alar defects. METHODS: Twenty-nine adult cadavers were anatomically used for vascular investigation of the frontal, nasal and temporal regions by injecting a dye into the main vessels. Based on the anatomical study in these regions, a retro-grade auricular flap was designed for repairing alar defects in 16 patients. RESULTS: The blood supply of the auricle could be nourished by the inner carotid artery system from the supratrachlear artery and supraorbital artery through the frontal arterial anastomotical network into the frontal branch of the superficial temporal artery. It is then passing the main trunk of the superficial temporal artery into the ear area through the auricular branches of the superficial temporal artery. The retro-grade auricular island flap could be formed by basing the supratrachlear artery and the supraorbital artery through the vascular network between the superficial temporal artery and the supratrachlear artery or the supraorbital artery. Sixteen patients with large alar defects and half-sized nasal defects were successfully repaired by this technique. CONCLUSIONS: The retro-grade auricular island flap, based on the inner carotid artery system, could be a good and safe flap for repairing a large alar defect or half-sized nasal reconstruction. PMID- 12889188 TI - [Correcting the congenital ala defect of the facial cleft with revolving reposition]. AB - OBJECTIVE: To introduce the experience of correcting the congenital ala defect of facial cleft with the method of revolving reposition. METHODS: Five patients were treated with this method. Based on features of the defect, two different operations were employed with the medial crura or the lateral crura of the ala as the revolving pivot. RESULTS: All the operations obtained good results. CONCLUSION: The operation of revolving reposition for congenital ala defect is a simple practical method with satisfactory results. PMID- 12889189 TI - [Reconstruction of thoracic and pericardial defects and consequent cardiac exposure in a thoracoventropagus following the separation surgery]. AB - OBJECTIVE: To explore and apply a reconstructive technique for thoracic and pericardial defects and cardiac exposure in a thoracoventropagus following the separation operation. METHODS: The separation operation was performed by the abdominal, the cardiothoracic and the plastic reconstructive surgeons in three teams. Nonbiological material was used in infant A to reconstruct the defective his thorax and pericardium resulted from the separation surgery. During the operation, infant B was protected with emphasis, preserving larger amount of thoraco-abdominal tissue to facilitate one-stage repair by direct suturing. Multiple tongue-shaped large skin flaps were designed to repair the huge soft tissue defects. The wound was closed in one stage. RESULTS: Both of the infants survived after the surgery. Infant A with severe defect of his anterior thorax and abdominal and consequent cardiac exposure was eventually cured in 3-year hospitalization. CONCLUSION: The repair and reconstruction of the defective tissues and organs in the thoraco-abdminal conjoined twins were highly difficult. The modern plastic surgical techniques used for the task were reliable, effective and successful. PMID- 12889190 TI - [Classification and treatment of asymmetrical breast deformity]. AB - OBJECTIVE: To evaluate the classification and the treatment of the asymmetrical breast deformity. METHODS: From April of 1997 to October of 2001, 90 patients with the asymmetrical breast deformity were undergoing in the study. The patients were classified and the results of the treatment were evaluated with the pre operative and post-operative photographs. RESULTS: The deformities of the asymmetrical breast could be divided into seven types according to the selected treatment techniques. The results were very satisfactory in 77 cases (87%), general satisfactory in 10 cases (11%) and unsatisfactory in 3 cases (2%). There were 54 cases (60%) with complete symmetry results, 32 cases (35.5%) with general symmetry and 4 cases (4.5%) with still asymmetry. CONCLUSION: The indications for treatment of the breast asymmetry deformities should be correlated with the classification so as to achieve more satisfactory results, in fact, it is still difficult to be a complete symmetry. PMID- 12889191 TI - [A conjunctival pulling-back technique for treatment of serious symblepharon]. AB - OBJECTIVE: To evaluate a new technique for treatment of the symblepharon caused by physical and chemical burn. METHODS: Thirty-eight patients with 40 eyes were undergoing the treatment. It was carried out to reconstruct the conjunctival fornix by pulling the remained conjunctive backward from the corneal limbus in serious stages. RESULTS: Thirty-eight patients (40 eyes) were treated with this technique, with the successful results of 13 patients (13 eyes) in 2 times, 19 patients (21 eyes) in 3 and 6 patients (6 eyes) in 1. Only one case was failure. CONCLUSION: The above mentioned technique could be a safe and effective way for treatment of symblepharen. PMID- 12889192 TI - [Experimental study of using xenologous sclera as wrapping materials for hydroxy apatite(HA)]. AB - OBJECTIVE: Use xenologous sclera (pig sclera) as wrapping materials for hydroxyapatite (HA) ocular implants in enucleated New Zealand white rabbits. METHODS: Ten rabbits were unilaterally enucleated and the sockets implanted with 10 mm HA spherical implants. All implants were wrapped with pig sclera. The rabbits were killed 4 weeks later and the sockets were examined clinically for inflammation and implant exposure. Exenterated specimens were assessed histopathologically for inflammation and vascularization. RESULTS: No clinical evidence of implant exposure or extrusion was detected in rabbits receiving pig sclera wrapped implants. Histopathologically the xenologous (pig) sclera displayed moderate inflammation, as well as maintain its lamellar arrangement without disruption. Fibrovascular ingrowth was complete in all implants. CONCLUSION: Xenologous (pig) sclera behaves similar clinically to homologous sclera as a wrapping material for spherical HA ocular implants in a rabbit model, and may represent a reasonable alternative to sclera in such procedures in humans. PMID- 12889193 TI - [The affect of the skin stretch on the biomechanical changes of skin]. AB - OBJECTIVE: To investigate the biomechanical changes in skin after employing the skin stretch. METHODS: Skin samples were took from the test group which was stretched for 7 days and control group without stretch respectively in the end of 1, 2, 3, 4, 6, 8 week. The biomechanical index were measured by tensiometer. RESULTS: The mean destroy stress, breaking load, stretch rate of the test group decreased obviously in the first week (the breaking load was increasing at the beginning) and the three index increased subsequently, they reached the climax during the fourth week and turned to normal subsequently to the level of some higher than control group and normal value in the sixth week. The stiffness of the test group reached the climax at the first week and than decreased gradually to the level of some higher than normal value and control group. The biomechanical changes of te control group was not evident by comparing with the test gropu. CONCLUSIONS: Skin stretch may injured the biomechanical property during the early time and turned to normal soon afterwards. The stiffness of the skin was increased and its elasticity was decreased after performing skin stretch. PMID- 12889194 TI - [Correction of nipple depression by using rhomboid de-epithelialized subcutaneous flaps]. AB - OBJECTIVE: To evaluate a new technique by using rhomboid de-epithelialized subcutaneous flaps for correction of congenital nipple depression. METHODS: Two de-epithelialized subcutaneous flaps were formed in the areola and placed into the base of the nipple after the nipple was elevated by releasing the fibrous tissue between depressed nipple and its base. The base was then tightened to reinforce the protrusion of the nipple and prevent the withdrawal of the nipple. RESULTS: Nine patients with 17 nipples were successfully treated with above technique between 1993 and 2001. Six patients were followed up for a period from 6 to 18 months. The results were satisfactory regarding to the shape and projection of the nipple. There was no visible scar and decreasing sensation of the nipple. One woman had children after the operation and fed well. CONCLUSION: The above mentioned technique may be an effective surgical procedure for correction of nipple depression with the advantages of preserving the lactiferous ducts, no visible scar and obtaining the sufficient protrusion and size of the nipple. PMID- 12889195 TI - [The reconstruction of neovascularization of posterior pharyngeal flap: an experimental study in dogs]. AB - OBJECTIVE: To elucidate the reconstruction of neovascularization that occurred in the superiorly and inferiorly based posterior pharyngeal flaps in different time postoperatively. METHODS: Ten mongrel dogs were randomly divided into two experimental groups, which were performed superiorly or inferiorly based posterior pharyngeal flap surgery respectively. Each group was then subdivided into five subgroups, and were sacrificed immediately after operation or on 3, 7, 30, 90 day postoperative respectively. Microangiography was used to exhibite the vessel. RESULTS: 1. The blood vessel reconstruction of the superiorly based posterior pharyngeal flap was more rapid compared with the inferiorly based flap. The 3-day flap has established an axial vascular network, which was mature on the 30-day flap. The superiorly based posterior pharyngeal flap was mainly supplied by the pedicle. 2. The blood vessels reconstruction of the inferiorly based posterior pharyngeal flap was firstly occurred in the pedicle and apex of the flap, which grew slowly to the middle of the flap. The inferiorly based posterior pharyngeal flap was supplied by the pedicle and the soft palate. A mature axial vascular network was exhibited on the 90-day flap, which was not mature on the 30 day flap. CONCLUSION: Both superiorly and inferiorly based posterior pharyngeal flap can establish an axial vascular network and gain ample blood supply. PMID- 12889196 TI - [The volume change of micronized acellular dermal matrix after injection]. AB - OBJECTIVE: To explore a new injectable material. METHODS: Micronized swine acellular dermal matrix was injected into the ear of rabbits. The augmentation was observed and measured with pathological and transmission electron microscopic technique. RESULTS: There were fibroblasts and capillaries in the injected material. The absorption rate of injected acellular dermal matrix was 26.93% in the experimental group at 48 weeks after injection. CONCLUSION: The acellular dermal matrix is an ideal injectable material for lower absorption and easy usage. PMID- 12889197 TI - [Ezras Asratovich Asratian (1903-1981)]. PMID- 12889198 TI - [The path of E. A. Asratian in science]. PMID- 12889199 TI - [Conditioned switching-over and cognitive set]. AB - The importance of Asratyan's conditioned switching-over phenomenon as an experimental model for investigation of the control function of the inner states (sets) in behavior is discussed. It is argued that experimental investigation of the animal conditioned switching-over phenomenon at the novel cognitive stage of exploration of neural mechanisms of behavior using the modern conceptual models of brain operation and conceptual apparatus can be a promising approach to understanding of the physiological basis of human unconscious cognitive sets and their role in perception of external signals and organization of voluntary behavior. PMID- 12889200 TI - [Evolutionary aspects of compensation of functions of the damaged spinal cord]. AB - In animal experiments with spinal lesions, phylo- and ontogenetic features of development of plasticity in the CNS of mammals and high reliability of the CNS have been demonstrated. It has been shown that possibilities of compensation processes and the upper limit of plasticity increase in phylogeny. Much attention has been attached to the higher regions of the CNS and hypothalamus as well as to ecological and biological characteristics of animals. Changes in the structure and functions of the damaged spinal cord have been studied. The significance of sympathetic innervation and involvement of ATP and ATPase in the spinal cord functioning after injury have been demonstrated. The article reviews the investigations of the role of the cicatrixce and some endocrine glands (adrenal glands, pancreas, and thyroid gland) performed at the laboratory. In phylogeny, high plasticity has been demonstrated at early ontogenetic stages. Enzymes have been shown to facilitate recovery of spinal functions after injury. PMID- 12889201 TI - [Interaction between neurons of the frontal cortex and hippocampus during the realization of choice of food reinforcement quality in cats]. AB - Six cats were subjected to the procedure of appetitive instrumental conditioning (with light as a conditioned stimuls) by the method of the "active choice" of reinforcement quality. Short-delay conditioned bar-press responses were rewarded with bread-meat mixture, and the delayed responses were reinforced by meat. The animals differed in behavior strategy: four animals preferred the bar-pressing with a long delay (the so-called "self-control" group), and two cats preferred the bar-pressing with a short delay (the so-called "impulsive" group). Multiunit activity in the frontal cortex and hippocampus (CA3) was recorded via chronically implanted nichrome wire semimicroelectrodes. An interaction between the neighboring neurons in the frontal cortex and hippocampus (within local neural networks) and between the neurons of the frontal cortex and hippocampus (distributed neural networks in frontal-hippocampal and hippocampal-frontal directions) was evaluated by means of statistical crosscorrelation analysis of spike trains. Crosscorrelations between neuronal spike trains in the delay range of 0-100 ms were explored. It was shown that the number of crosscorrelations between the neuronal discharges both in the local and distributed networks was significantly higher in the "self-control" cats. It was suggested that the local and distributed neural networks of the frontal cortex and hippocampus are involved in the system of brain structures which determine the behavioral strategy of animals in the "self-control" group. PMID- 12889202 TI - [Interhemispheric asymmetry in the functional organization of the prefrontal cortex in animals of different typologic groups]. AB - Rats allocated to groups by the method of "emotional resonance": rats which did and did not escape crying of a partner (A- and E-groups, respectively). Unit activity in the right and left prefrontal brain cortex (PFC) was recorded in these rats. The recorded neurons neurons were divided in two groups according to their reaction to a change in the level of food motivation. The so-called D neurons decreased their activity after feeding of animals after a 24-hour food deprivation and the other group (I-neurons) increased its firing rate rate in this situation. It was shown that hemispheric distributions of D- and I-neurons are different in selected rat groups. In E-rats the I-neurons substantially predominated in the left hemisphere, whereas the D-neurons were more frequently recorded in the right one. No such asymmetry was observed in A-group of rats. During intracranial stimulation of emotionally positive brain structures I neurons increased their firing rate, predominantly, in the left hemisphere, whereas during intracranial emotionally negative stimulation activation of the D neurons predominated at the right. Features of the observed functional interhemispheric asymmetry of prefrontal cortex in A- and E-groups of rats were explained by differences in the interaction between hemispheres and dissimilar activation control. PMID- 12889203 TI - [Features of learning voluntary control of posture after pyramidal and nigrostriatal lesions]. AB - The study was aimed at investigation of a deficit of learning the center-of pressure voluntary control in patients with lesions of corticospinal and nigrostriatal systems. Thirty three patients with Parkinson's disease and 20 patients with hemiparesis after cerebrovascular accidents in the MCA participated in the investigation. The subjects stood on a force platform and in the form of a computer game were trained to match the projection of the center of pressure (a cursor) with a target on the screen under the visual feedback control. Two different postural tasks were presented. In the first task the direction of the center-of-pressure shift was not known before, so the subject learned the general strategy of the center-pressure control. In the other task a precise postural coordination should be formed. The voluntary control of the center-of-pressure position was found to be impaired in both groups of patients. In the task of moving the center of pressure in various directions (general strategy), no differences in the initial deficit of the task performance were found between the groups, but the learning was more efficient in the group of hemiparetic patients. However, in the task with precise postural coordination, despite the greater initial deficit in the parkinsonian patients, the learning in this group of patients was substantially more efficient than in hemiparetic patients. The results suggest both common and different features of the involvement of the corticospinal and nigrostriatal systems in learning voluntary control of posture. PMID- 12889204 TI - [Functional asymmetry of the frontal cortex and lateral hypothalamus of cats during food instrumental conditioning]. AB - The synchronism and latency of auditory evoked potentials (EP) recorded in symmetric points of the frontal cortex and lateral hypothalamus of cats were measured at different stages of instrumental food conditioning and after the urgent transition to 30% reinforcement. Correlation coefficients between EPs in the cortex and hypothalamus were high (with left-side dominance) at the beginning of the experiments, when food motivation was high, and during the whole experiments in cases of high-probability of conditioned performance. Analysis of early positive P55-80 EP component showed that at all conditioning stages the peak latency of this component was shorter in the left cortical areas than in symmetrical points, whereas in the hypothalamus the shorter latency at the left side was observed at the stage of unstable conditioned reflex, and at the stage of stable reflex the latency of the studied component was shorter at the right side. During transition to 30% reinforcement, the latency was also shorter in the right hypothalamus. It is suggested that the high left-side correlation between the hypothalamus and cortex was associated with motivational and motor component of behavior rather than reflected the emotional stress induced by transition to another stereotype of food reinforcement (30%). PMID- 12889205 TI - [Analysis of the effect of a training regime for the correct choice of matched learning in gray crows]. AB - It was shown that a large set of training stimuli promotes abstract concept learning. These experiments were designed to assess whether an application of a large set of training stimuli would facilitate matching learning in crows. Four hooded crows were trained with a set of 72 unique combinations of stimuli in two alternative simultaneous matching tasks with stimuli of three different categories: achromatic color (white, light-grey, dark-grey, and black), shape (Arabic numerals from 1 to 4 used as visual shapes only), and number of elements (heterogeneous graphic arrays from 1 to 4 items). Although the performance of all crows was significantly above chance (p < 0.01) in some 72-trial blocks, birds were unable to establish matching and to reach the criterion of learning 80% correct or better over 72 consecutive trials) in 5184 trials. Thus, the modified training procedure was less efficient than the training technique previously used (successive cyclic repetition of three small sets of training stimuli), which allowed four of six crows to acquire the matching rule after 1780, 2360, 3830, and 5260 trials [4,9]. PMID- 12889206 TI - [From physiological theory to psychological facts (on the centenary of I. P. Pavlov's speech in Madrid)]. AB - Pavlovian theory of conditioned reflexes and Ukhtomsky's theory of dominanta are considered as the most important biological concepts of the XX century. They have laid the foundation of the modern views of the adaptive and active character of the nervous activity underlying the integral behavior in the environment. Approach to a burning problem of neurophysiological mechanisms of goaldirected behavior is impossible without a comprehensive study of works of both these schools. They have formed not only the methodological basis of modern investigations but opened a new avenue of attack. PMID- 12889207 TI - [Positive potentials of the human brain at different stages of preparation of a visually triggered saccade]. AB - The EEG of 10 right-handed subjects preceding saccades with mean values of latent periods were selected and averaged. Two standard paradigms of presentation of visual stimuli (central fixation stimulus-peripheral target succession): with a 200-ms inerstimulus interval (GAP) and successive single step (SS). During the period of central fixation, two kinds of positive potentials were observed: fast potentials of "inermediate" positivity (IP) developing 600-400 ms prior to saccade onset and fast potentials of "leading" positivity (LP), which immediately preceded the offset of the central fixation stimulus. Peak latency of the LP potentials was 300 ms prior to saccade onset in the SS paradigm and 400 ms in the GAP paradigm. These potentials were predominantly recorded in the frontal and frontosagittal cortical areas. Decrease in the latency by 30-50 ms in the GAP paradigm was associated with more pronounced positive potentials during the fixation period and absence of the initiation potential P-1' (or decrease in its amplitude). The obtained evidence suggest that the fast positive presaccadic potentials are of a complex nature related to attention, anticipation, motor preparation, decision making, saccadic initiation, and backward afferentation. PMID- 12889208 TI - [Changes in the brain's electrical activity during habituation to verbal stimulus in subjects with high and low levels of personal anxiety]. AB - Brain electrical activity during habituation to repeated verbal stimulus was recorded in subjects with high and low levels of personal anxiety. During habituation of subjects with low anxiety, the spectral power of EEG alpha and theta bands increased in the frontal cortical areas. Such a reaction was not observed in subjects with high anxiety level. Moreover, during habituation traces, the latency of P300 of the auditory evoked potential increased. This parameter did not depend on anxiety level. The amplitude of P300 in the right temporal region was shown to be related to anxiety level: it increased after habituation in subjects with low anxiety and did not change in subjects with high anxiety. PMID- 12889209 TI - [Effects of repeated experience of aggression on aggressive motivation and development of anxiety in male mice]. AB - The sensory contact technique allows an aggressive type of behavior to be formed as a result of repeated experience of social victories in daily agonistic confrontations. In mice of the low-aggressive and high-emotional CBA/Lac strain repeated positive fighting experience increases plus-maze anxiety. Behavioral reactivity of males to other conspecifics in the partition test (which measures aggressive motivation) significantly rises. It is concluded that repeated experience of aggression provokes the development of anxiety in male mice. The results suggest that level of anxiety and its behavioral realization depend on duration of aggressive experience and genetic strain. PMID- 12889210 TI - [EEG spatial characteristics after intense exercises]. AB - Subjects performed intense exercises in a teadbahn under conditions of high ambient temperature and humidity. Spatial characteristics of the EEG were estimated. The information value of the time course of the combined index, characteristics of spatial synchronization and entropy of cortical biopotentials as a test for brain reaction to a load and recovery of its functional state was shown. The significance of non-linear relations between brain potentials in the activation of the adaptive and compensation processes was mentioned. PMID- 12889211 TI - [Orienting behavior of female white rats in estrus cycle and its dependence on handling]. AB - Characteristics of orienting behavior of intact and handled female white rats were studied in the "open field" test. Experimental group of animals were formed according to stages of estrous cycle: (1) diestrus, (2) proestrus, (3) estrus, and (4) metestrus. The stage of the cycle was determined by vaginal smears. Over the period of 10 days rats were handled daily for 5 minutes. No behavioral changes over the course of estrous cycle were found in intact rats. Handling revealed some behavioral features related to the levels of steroid hormones in reproductive cycle. Most prominent changes in orienting behavior were observed at transition from estrous to metestrous. At the stage of estrous motor activity was maximal and grooming was minimal. The maximal contrast in the structure of orienting behavior was observed between estrous and diestrous stages. PMID- 12889212 TI - [Duration of changes in electrical characteristics of command neurons after defensive conditioning in snails]. AB - Electrical characteristics of snail command neurons were studied during and after defensive conditioning. Tapping on the shell was used as a conditioned stimulus. A light air blow into the lung cavity orifice (pneumostome) was used as an unconditioned stimulus. The conditioned defensive reflex is known to be retained for 40 days. We have shown earlier the decrease in membrane and threshold potentials of command neurons after defensive conditioning (Gainutdinov et al., 1996). In these experiments it has been found that the decreased level of membrane and threshold potentials are maintained during 40 days after defensive reflex conditioning. PMID- 12889213 TI - The assessment of the patient with systemic sclerosis. Introduction. PMID- 12889214 TI - Assessment of skin, joint, tendon and muscle involvement. AB - This report makes recommendations for standardized techniques of data gathering and collection regarding: 1) skin involvement 2) joint and tendon involvement, and 3) involvement of the skeletal muscles. The recommendations in this report derive from a critical review of the available literature and group discussion. Committee recommendations are considered appropriate for descriptive clinical investigation, translational studies and as standards for clinical practice. Skin involvement should be assessed using the modified Rodnan skin score. Joint involvement, when symmetric synovitis is present, could be best assessed by the DAS-28 as is utilized in rheumatoid arthritis. Clinical assessment should include a routinized evaluation for the presence and number of palpable tendon friction rubs. Muscle involvement should be screened for by performance of the serum creatine phosphokinase assay and assessment of proximal weakness. More specific testing including EMG, magnetic resonance imaging and muscle biopsy should be employed in those patients with clinically significant myopathy only. PMID- 12889215 TI - Assessment of vascular involvement. AB - Our objective was to identify a core set of variables for the assessment of vascular involvement in scleroderma that is simple, reproducible, and reflects the presence of vascular disease in SSc. To do so we carried out an extensive literature review of published studies relating to the assessment of vascular involvement in SSc, i.e. studies dealing with clinical parameters, functional vascular studies, cold presser testing, nailfold capillary microscopy and circulating vascular markers. After extensive review of published studies and critical assessment of proposed vascular parameters, the subcommittee endorsed what it considers to be the minimal requirements for the documentation of vascular involvement in SSc. The core set variables include two parameters: Raynaud's phenomenon and digital ulcers. This set is simple, reproducible and should be included in the assessment of SSc patients in clinical investigational studies. The subcommittee also however recognizes that there exists a promising set of vascular variable that still needs further investigation. PMID- 12889216 TI - Assessment of gastrointestinal involvement. AB - The purpose of this paper is to identify a list of clinical, laboratory and instrumental tools suitable to assess the presence of gastrointestinal involvement in SSc patients to be included in clinical investigational studies. The pertinent literature was reviewed to select those variables which have been demonstrated to be valid, reliable and feasible. A minimal core set of variables has been identified to be used in clinical investigation for the assessment of esophagus, stomach, small intestine, colon and anorectum involvement in scleroderma patients. PMID- 12889217 TI - Assessment of lung involvement. AB - The aim of the subcommittee was to identify a core set of feasible variables reflecting the occurrence of interstitial and/or vascular lung disease. After extensive review of published studies and critical assessment of candidate variables, the subcommittee identified the minimal requirements to assess lung disease. Two core sets of variables are provided: the first concerns interstitial lung disease; the second pulmonary vascular disease. PMID- 12889218 TI - Assessment of heart involvement. AB - Cardiac involvement frequently occurs in systemic sclerosis (SSc), contributing to the occurrence of symptoms, namely dyspnoea, fatigue, palpitations, and in some instances to the clinical evolution and prognosis of the disease. A thorough baseline screening of heart functioning and appropriate follow-up monitoring is therefore mandatory in all SSc patients. This consists of various simple, non invasive ambulatory diagnostic procedures (visit, electrocardiogram, chest X-ray, Doppler-bidimensional echocardiogram), which provide information on the presence of rhythm and conduction disturbances, cardiac morphology and function, as well as on the possible presence of pulmonary hypertension (PH). When needed, added tests may be carried out, including long-term ambulatory electrocardiographic recording, assessment of cardiopulmonary performance by the six-minute walking test or cardiopulmonary stress test, cardiac catheterization (mandatory to confirm and better estimate PH), cardiac magnetic resonance imaging, and nuclear studies of myocardial function and perfusion. PMID- 12889219 TI - Assessment of kidney involvement. AB - Scleroderma renal crisis (SRC) represents the classic manifestation of kidney involvement in SSc. It particularly occurs in patients with early, rapidly progressive, diffuse skin involvement. Its detection requires the assessment of a few core set variables: arterial blood pressure, serum creatinine, and urinalysis. In clinical investigations SSc patients developing arterial hypertension after the disease onset (new onset hypertension) without SRC should also be reported. PMID- 12889220 TI - Non organ based laboratory markers in systemic sclerosis. AB - Candidate markers for endothelial cell, lymphocyte and fibroblast function in systemic sclerosis have been reviewed. Autoantibody measurement and general inflammatory markers were considered the minimum requirements, with other markers needing further evaluation. PMID- 12889221 TI - Assessment of disease activity. AB - The objective of this study was to select a simple, feasible and validated core set of variables for the assessment of disease activity in systemic sclerosis. The subcommittee members extensively reviewed the studies in which the disease activity had been assessed, and analysed the construct validity and the feasibility of the criteria used in each study. The subcommittee members agreed on the possible use of either the European Scleroderma Study Group (ESSG) criteria for disease activity in SSc or the evaluation of selected cytokines and cell activation markers, found to be associated with distinct organ system involvement in recent studies. Despite the potential usefulness of cytokines and activation markers, most participants in the plenary session retained that their evaluation is not feasible in a number of centres, and their construct validity has not yet been assessed. The ESSG criteria for disease activity can be used for the time being to assess disease activity in clinical investigation studies as well as in clinical practice. PMID- 12889222 TI - Assessment of disease severity and prognosis. AB - The Subcommittee members initially agreed on the concepts of disease activity, damage and severity, defining severity as the total effect of disease on organ function. It was decided to start with the assessment of severity using the Medsger's severity scale. A revised version of this scale was constructed. The rationale for the exclusion of other variables was provided. PMID- 12889223 TI - Conclusion and identification of the core set of variables to be used in clinical investigations. PMID- 12889224 TI - Manual of signs, symptoms, methods and procedures for the assessment of the patient with SSc. PMID- 12889225 TI - Diabetes: get a clearer picture. PMID- 12889226 TI - Bringing HOPE for diabetes treatment to China. PMID- 12889227 TI - Digene's chairman/CEO Evan Jones. Interview by Carren Bersch. PMID- 12889228 TI - From peapods to laboratory medicine: molecular diagnostics of inheritable diseases. PMID- 12889229 TI - Deliver the lesson now: just-in-time training. PMID- 12889230 TI - Patient's right to choose raises points with common grounds. PMID- 12889231 TI - Clinical laboratories and the shaoe of Medicine reform . PMID- 12889232 TI - Wholesale to retail shift requires providers to adapt. PMID- 12889233 TI - ER screening brings bad press to hospitals. PMID- 12889234 TI - Cogent distinguishes self in rapidly growing field of hospitalist care. PMID- 12889235 TI - Report: sale of office buildings can add liquidity, fund other needs. PMID- 12889236 TI - Solucient report predicts pediatric admissions to jump 20% by 2027. PMID- 12889237 TI - Health Affairs study shows private health spending rose again in 2002. PMID- 12889238 TI - The ever changing face of laparoscopic surgery in South Africa. PMID- 12889239 TI - Non-operative management of splenic injury in adults--current management status and controversies. PMID- 12889240 TI - Evidence-based medicine--the two-edged sword. PMID- 12889241 TI - Reduction mammoplasty in cases of giant fibroadenoma among adolescent females. Case reports and literature review. AB - Giant tumours of the breast cause massive enlargement of the breast resulting in asymmetry. Simple excision of the tumour fails to restore symmetry, and in some cases leaves the patient with a secondary deformity. We present two cases of giant fibroadenoma in adolescent females. Both patients underwent a one-stage excision of the giant fibroadenoma and reduction mammoplasty. Relevant operative details and results are presented together with a review of the literature. PMID- 12889242 TI - Open mesh-plug technique in inguinal hernia repair--short-term results. AB - OBJECTIVE: To evaluate the safety, technical problems, early complications and short-term results of the mesh-plug technique in primary and recurrent inguinal hernia repair. DESIGN: Prospective study. SETTING: Armed Forces Hospital, Southern Region, Khamis Mushayt, Saudi Arabia. SUBJECTS: Two hundred and forty four patients with 256 hernias were admitted through a specialised hernia clinic between May 1997 and June 1998. There were 236 men and 8 women, with a mean age of 50 years (range 17-90 years). INTERVENTIONS: Repair of the inguinal hernia using the mesh-plug technique. RESULTS: Concomitant medical problems were present in 7.3% of patients. The size of the hernia was assessed preoperatively and recorded as either inguinal (73%) or inguino-scrotal (33%). The site of the hernia was right in 55.7% of patients, left in 48% and bilateral in 4.9%. Furthermore, types and Gilbert's classification were assessed intraoperatively. The mean operative time was 41.61 +/- 10.59 minutes and the mean hospital stay was 1.14 +/- 2.63 days. No perioperative mortality occurred in this series. General complications were vomiting in 4 cases and 6 cases of chest infection. Local complications included 2 would infections (0.8%), 26 skin bruises (10.7%), 6 scrotal swellings (2.5%), 2 cases of cord thickening (0.8%) and 16 cases of postoperative neuralgia (6.6%). There were only 2 cases of recurrence (0.8%). CONCLUSION: Simplicity, safety, short operating time, and low rates of complications and recurrence were noted. The technique is promising and could be used for all types of primary and recurrent inguinal hernia as day surgery cases under local anaesthesia. PMID- 12889243 TI - Aortic arch branch variations--case report and arteriographic analysis. AB - OBJECTIVES: Non-recognition of a critical aortic arch branch variation at surgery, with fatal consequences, prompted a retrospective analysis of the prevalence of aortic arch branch variations in the South African population. METHODS: Three hundred and twenty aortic arch angiograms were analysed from existing records (January 1988--March 1998) of the vascular unit, Department of Surgery, University of Natal. RESULTS: Aortic arch branch variations were observed in 17 cases (5.3%). These variations were as follows: (i) 11 cases with two primary branches (3.4%); (ii) 1 case with three primary branches (0.3%); and (iii) 5 cases with four primary branches (1.6%). CONCLUSION: The clinical presentations which these variations give rise to are well known and may present during the first days of life or later in adulthood, or remain clinically silent. Non-recognition of the latter situation in the presence of vascular trauma may have fatal consequences. PMID- 12889245 TI - [The meaning of health]. PMID- 12889244 TI - Trauma in pregnancy. PMID- 12889246 TI - [Intentionality of nursing care of ulcerated tumors in terminal oncology patients]. PMID- 12889247 TI - [Nursing: between public health and the institution]. PMID- 12889248 TI - [Pregnancy in adolescence. Reflexions]. PMID- 12889249 TI - [HIV and AIDS: experiencing the symptoms. What cell count and viral burden do not tell us]. PMID- 12889251 TI - [Professional practice: why should nursing be chosen?]. PMID- 12889250 TI - [Significance of the tetanus vaccine update]. PMID- 12889252 TI - [Autonomy of the patient. A study for reflexion]. PMID- 12889253 TI - [The significance of assertiveness in the health team]. PMID- 12889254 TI - The evolution of emergency medicine in Ireland. AB - Irish emergency medicine is now evolving at a rapid pace. Having roots in a rich Irish medical history and different cultural and economic influences from the United States, the specialty has developed much differently than has American emergency medicine. The history of Irish emergency medicine is discussed and comparisons are made between an Irish and an American emergency department. PMID- 12889256 TI - Delaware's infant mortality rate exceeds national average. PMID- 12889255 TI - The social impact of medicine. Chapter 9. PMID- 12889257 TI - Protecting against SARS during equipment maintenance. AB - The recent outbreak of severe acute respiratory syndrome (SARS) raises questions about the appropriate infection control measures to use when conducting maintenance procedures on medical devices that may have been exposed to the SARS virus--that is, devices that have been used on, or located in the same room as, a patient with (or suspected to have) SARS. Although there is considerable information and guidance available on many aspects of SARS, very little exists on this particular topic. What's more, we have found significant differences in the perspectives and recommendations of some of the experts on the subject. This article provides guidance for clinical engineering, respiratory therapy, and other personnel involved in maintaining potentially SARS-exposed equipment. To derive our recommendations, we reviewed the material posted on the Web sites of the U.S. Centers for Disease Control and Prevention (CDC) and the World Health Organization (WHO), along with other information sources; we also discussed issues related to equipment use and servicing with experts outside ECRI. PMID- 12889258 TI - Mechanical ventilation of SARS patients. Safety issues involving breathing circuit filters. AB - Patients suffering from severe acute respiratory syndrome (SARS) sometimes require mechanical ventilation. This raises safety concerns about the SARS virus being carried by droplets that exit from the exhalation limb of the ventilator. ECRI recommends that breathing-circuit filters be incorporated in the exhalation limb of any ventilator used on a patient with SARS. In this article, we review the issues behind our recommendations and describe steps hospitals can take to prevent any adverse effects on ventilation. PMID- 12889259 TI - Automated external defibrillators (AEDs). AB - Automated external defibrillators, or AEDs, will automatically analyze a patient's ECG and, if needed, deliver a defibrillating shock to the heart. We sometimes refer to these devices as AED-only devices or stand-alone AEDs. The basic function of AEDs is similar to that of defibrillator/monitors, but AEDs lack their advanced capabilities and generally don't allow manual defibrillation. A device that functions strictly as an AED is intended to be used by basic users only. Such devices are often referred to as public access defibrillators. In this Evaluation, we present our findings for a newly evaluated model, the Zoll AED Plus. We also summarize our findings for the previously evaluated model that is still on the market and describe other AEDs that are also available but that we haven't evaluated. We rate the models collectively for first-responder use and public access defibrillation (PAD) applications. PMID- 12889260 TI - AEDs in your facility. A risk management perspective. AB - Recently, many hospitals have begun installing automated external defibrillators (AEDs) to help reduce cardiac arrest response times. The devices are being placed in areas where manual defibrillators generally cannot be deployed quickly enough to be effective. This Guidance Article is intended to help facilities that are trying to start an AED program. We discuss important factors to consider, and we outline a specific implementation model that can be followed. In addition, the article covers liability issues that may arise, regardless of whether your facility has an AED program in place. For model-specific information, refer to the Evaluation on AEDs earlier in this issue. PMID- 12889261 TI - Worn AC wire insulation on examination lights causes device failure and shock hazard. PMID- 12889262 TI - Some infant ventilators may shut down without backup power. PMID- 12889263 TI - NIBP hoses do not connect securely with GE Dinamap monitors. PMID- 12889264 TI - Frequent failures of bed-exit devices on Hill-Rom 153 Advance Series beds. PMID- 12889265 TI - [Camptodactylies]. AB - Camptodactyly is a permanent non-traumatic flexion contracture at the proximal interphalangeal joint, involving most of the time the fifth finger. This condition is rare (incidence is inferior to 1%). Most of the cases are sporadic, although some authors have traced the transmission through several generations. Two types of camptodactyly have been described, depending on the age of onset (within the first year of age or in adolescent). This condition can also be described as "static" or "dynamic", depending on the effect of MP flexion on the PIP extension lag. Many anatomical abnormalities have been evoked as potential aetiology for camptodactyly, including variations in the lumbrical or flexor superficialis origin or distal insertions. However, after a few years without treatment, permanent flexion position of the joint adds its own effects and leads to joint contracture, regardless of the aetiology of camptodactyly. Treatment is required for this condition for extension deficit greater than 30 degrees. The first step will always consists of dynamic splinting of the PIP joint. If improvement is not obtained through dynamic splinting alone, surgery can be considered. Correction of a well defined anatomical abnormality is part of the treatment. When required, it will be associated to correction of joint and soft tissue contracture. PMID- 12889266 TI - ["Square sail" flap in median nerve lesions at the wrist. Anatomical elements and review of twenty operated cases]. AB - INTRODUCTION: Following an anatomical study on the vascular supply of the upper limb, we propose a new adipo-fascial flap at the wrist. The fat pad of pronator quadratus lies over the muscle and is vascularised by a recurrent branch arising from the anastomotic "cross-road" of the anterior interosseous, radial and ulnar arteries, at the radio-carpal joint. The peripheral extensions of the adipose tissue have to be anchored to the palmar fascia of the muscle in order to hoist the flap as a "square sail". The flap is then distally rotated in order to cover the traumatized median nerve segment at the wrist, for 2.5-3 cm of length. METHODS: The authors present a review of the clinical applications (1995-2001) on painful neuromas of the median nerve at the wrist, where traumatic and iatrogenic injuries frequently occur. The surgical procedure consists of external neurolysis followed by coverage of the nerve using this vascularised flap. RESULTS: The results confirm the usefulness of vascularised fat flaps in creating an optimal perineural environment in terms of biological and mechanical quality. The advantage offered by this particular local flap is that it can be speedily raised. Because of its limited length the flap is only able to cover the median nerve at the level of the carpal tunnel: its indications are therefore very selective but not rare, due to the frequency of median nerve injuries at this level. DISCUSSION: Clinical results were very satisfactory: in the twenty cases reviewed, resolution of the symptoms of algodystrophy and causalgia was consistently observed, sometimes to an extraordinary degree, and associated with an improvement in the neurological deficit. PMID- 12889267 TI - [Multiple peripheral nerve tumors: update and review of the literature]. AB - Multiple tumours of the peripheral nerves are seen only in neurofibromatosis. They are hereditary. They present and develop in a variety of different ways. Three main groups are distinguished: von Recklinghausen neurofibromatosis or type 1; bilateral acoustic neurofibromatosis or type 2 and schwannomatosis recently defined as type 3. The aim of this study was to clarify the clinical outcome of neurofibromatosis. The diagnosis is made purely on clinical grounds. Cranial MRI and slit lamp examination are useful for classification. Surgical management for peripheral nerve tumours is similar. Any new and rapid change noted at clinical examination (increase in volume, pain or neurological deficit) requires surgery because of potential malignant transformation of the neurofibroma into neurofibrosarcoma (type 1 only). The definitive treatment depends on the resectable character of the tumour which is usually only known after epineurotomy under operating microscope. In the event of resectable tumour (schwannoma) enucleation must be performed, preserving nerve continuity. In the event of unresectable tumour (neurofibroma), tumour resection is impossible without sacrificing nerve tissue. An epineurotomy must be performed. It prevents further deterioration. Interfascicular biopsy confirms the histological type. Our results are similar to those in other recorded studies. The unpredictable clinical course of neurofibromatosis makes prolonged follow-up mandatory. PMID- 12889268 TI - [A 36 month prospective study of 12 plate osteosyntheses of distal radius fractures]. AB - INTRODUCTION: We performed a prospective study of 12 intra-articular distal radial fractures. The fractures were treated with an anterior plate together with posterior pinning. The aim of the study was to examine whether it was technically feasible to fix such fractures using this technique and to assess the stability of the fixation at follow up. METHOD: Twelve articular fractures were included with a mean age of 47.1 years. All patients were followed up from first admission to the 36th postop month with clinical and radiological checks. RESULTS: At follow up, we found 8 good and excellent clinical results, 3 fair and 1 poor. The combination of an anterior plate with posterior pinning allowed a good initial reduction but could not protect from secondary displacement particularly in the presence of a comminuted metaphysal fracture. DISCUSSION: This procedure gave clinically satisfactory results but did not control ulnar variance. This procedure should be combined with bone grafting in complex metaphysal fractures and epiphysal screws should be used whenever possible. PMID- 12889269 TI - [Partial wrist prosthesis: concept and preliminary results in 13 cases]. AB - The author introduces a novel concept, namely the partial wrist prosthesis. This new technique combines a proximal row carpectomy with resurfacing of the lunate fossa of the radius and the head of the capitate. Results of the first 13 cases are reported with a mean follow-up of 32.7 months (13-72 m). This prosthesis can be used in degenative arthritis of the wrist instead of total fusion. PMID- 12889270 TI - [Resection of the first row of carpal bones: post-traumatic wrist and Kienbock's disease]. AB - This study reports the outcomes of 27 proximal row carpectomies for stage II (Watson) scapholunate--(10 Slac) and scaphoid non union--(8 Snac) advanced collapse and stage III (Lichtman) Kienbock's disease (9 cases) followed for an average of respectively 72 and 50 months. Following surgical treatment, more than 80% of patients in both groups were pain free. The total arc of motion averaged 67 degrees (unchanged), for the post-traumatic arthritis, and 59 degrees for Kienbock's, a decrease of 17%. Grip strength averaged a 17% increase in comparison to the opposite size for both groups. Proximal row carpectomy showed a high degree of patient satisfaction and is a motion-preserving and grip preserving procedure used in stage II post-traumatic arthritis but Lichtman III stage in Kienbock's disease had a comparatively poor result. Proximal row carpectomy simplifies the structure of the radiocarpal joint. When the cartilage on the capitate head and the radial lunate facet are not worn, this procedure provides a good result with respect to pain, range of motion and strength which is stable with time. PMID- 12889271 TI - [Conservative surgery in Kienbock's disease with perilunate arthrosis: articular resurfacing using resected carpal bones]. AB - We present 2 cases of Kienbock's disease in which peri-lunate chondral lesions contra-indicated classical procedures such as proximal row carpectomy or 4 corner arthrodesis. A partial carpectomy provided us with an osteochondral graft, used to resurface the areas of chondral damage over the capitate or on the radius. The clinical and radiological result was stable at a follow-up of 3 and 6 years. In the first case, degenerative changes over the head of the capitate contra indicated proximal row carpectomy. The lunate was removed and the proximal 2/3 of the scaphoid were shifted medially and fused in a "four-corner"-like arthrodesis. In the second case, the lunate fossa on the radius was damaged. The proximal row was excised and an osteochondral graft was harvested from the triquetrum. This was used to replace the lunate fossa on the radius. This new concept of a "carpal bank" has enabled us to extend the classical indications for proximal row carpectomy and four-corner arthrodesis. It makes it possible to withdraw the limits of conservative wrist surgery in Kienbock's disease and we believe this concept could also be extended to similar situations of localized chondral damage in small joints. PMID- 12889272 TI - [Giant cell tumor of the third metacarpal bone: a case report]. AB - A case of giant-cell tumor of the third metacarpal bone is reported, treated with resection and reconstruction by iliac graft. At a 5 years follow-up, the cosmetic and functional results of the hand are good. PMID- 12889273 TI - [Venous drainage in retrograde pedicle flaps: experimental study in rats]. AB - This study was performed to investigate the venous drainage in reverse island groin flaps in a rat model. Two groups of 10 rats were studied. All rats of group A had a groin reverse flap with a complete pedicle (artery and venae comitantes). In rats of group B, an arterial groin reverse flap (artery without venae comitantes) was performed. For the two groups, the perivascular tissue was excised. Nine flaps in the group A and seven flaps in the group B, survived without partial or complete necrosis. Microscopic examination showed venous dilatation in the two groups. There was no significant difference between the two groups. These results confirm that venous drainage of the arterial reverse flow flap without venae comitantes is performed by venae arteriosa. However, venae comitantes probably ensure venous drainage when they are respected. PMID- 12889274 TI - [Surgical correction of fifth finger permanent abduction by tenodesis. Preliminary cadaver study]. AB - Permanent abduction of the little finger can be responsible for daily embarrassment in patients with an ulnar nerve palsy. To correct this deformity, active transfers are usually performed utilising the extensor tendons of the hand. Because of the anatomical variability of the extensor system of the hand, these active transfers can be responsible for postoperative loss of full extension of the little finger. Analysis of the orientation of the forces generated by these transfers shows that they are only weak adductors. A surgical technique using tenodesis is proposed in this preliminary study. This tenodesis has the objective of increasing the adductive forces on the little finger without an extensor tendon transfer. The advantages and disadvantages of this technique are discussed. A clinical evaluation will be undertaken at a later date to confirm the reliability of this technique. PMID- 12889275 TI - More hospitals opting to lease equipment. PMID- 12889276 TI - Bandage price outlook mixed. PMID- 12889277 TI - Will new FDA rules hasten device approval? PMID- 12889278 TI - Planner must be good at crisis management. PMID- 12889279 TI - Reducing ophthalmic drug-related injuries in older patients. PMID- 12889280 TI - Catch me if you can. PMID- 12889281 TI - The use of megadose corticosteroids in traumatic optic neuropathy: case studies. PMID- 12889282 TI - Selecting eyeglasses: how not to make a spectacle of yourself. PMID- 12889284 TI - Yellow fever resurgent. PMID- 12889283 TI - Hepatitis A virus infections in injecting drug users. PMID- 12889285 TI - The Communicable Disease Surveillance Centre 1977-2002: an overview. AB - The Communicable Disease Surveillance Centre (CDSC) is now firmly established within the new Health Protection Agency (HPA), which was inaugurated on 1st April 2003. In 2002, CDSC celebrated its 25th anniversary. Its achievements over those 25 years, a period when it evolved from a small unit with three staff to an international centre of excellence with over 250 staff, are recalled in this paper. The development of the Centre is reviewed, as are the ways in which it identified and responded to changing patterns of communicable disease over the quarter century. The considerable benefits of placing CDSC within the Public Health Laboratory Service are demonstrated through several examples. The fact that the expansion of CDSC took place during an era when public expenditure was constrained indicates that government valued the service it provided. The elements required and put in place for effective control and prevention of communicable diseases are highlighted and several themes emerge including the close working relationship needed between the various agencies and disciplines, particularly those between epidemiologists, microbiologists and clinicians. Another theme is the commitment to training, (which is necessary to ensure that work is practiCed to the highest standards), as well as to innovation through research and development. A rigorous approach to field investigation is essential, not just to enable the most appropriate control measures to be applied but to increase the knowledge base on infections and the means of prevention. Good surveillance and reference microbiology stand out as the vital components needed to produce the timely and precise information required to influence practice and policy. PMID- 12889286 TI - Changing epidemiology of hepatitis A: should we be doing more to vaccinate injecting drug users? AB - Since 2001 there have been significant outbreaks of hepatitis A virus (HAV) across South Yorkshire, largely in intravenous drug users, and HAV infection has been reported to be an increasing problem in England and Scotland during this time. This paper reports a brief investigation to clarify current HAV epidemiology in England and Wales. The epidemiology of HAV in England, but not yet Wales, has recently changed. Laboratory reports now show that most cases are occurring in young adults, mainly young men, and that the commonest reported risk group is injecting drug users. That cases may now be concentrated in injecting drug users is supported by reports from consultants in communicable disease control (CsCDC). These detail fourteen outbreaks in England in 2002 alone, all involving injecting drug users. Links to prisons and to the homeless, usually those in hostels, were also common. A combined Hepatitis A/B vaccine is readily available and we recommend that this now be used to extend the national immunisation programme against Hepatitis B in injecting drug users to include HAV. PMID- 12889287 TI - Outbreak of hepatitis A infection among intravenous drug users in Suffolk and suspected risk factors. AB - A prolonged outbreak of hepatitis A infection amongst drug users in Suffolk prompted a study of the natural immunity against hepatitis A in this population, and a retrospective analysis of the relationship between specific drug-taking behaviours and the risk of hepatitis A infection. Prior to the outbreak, age specific seroprevalence of hepatitis A IgG in drug users was similar to that amongst blood donors in the region. Of those without effective immunity, intravenous drug users, multiple drug users and those injecting frequently were more likely to have developed hepatitis. The reported frequency of equipment sharing and the number of injecting partners were not related to the risk of infection. The potential for blood-to-blood, and a suggested faecal-blood transmission were considered to be important in propagating the outbreak in this population. We suggest that a single dose of hepatitis A vaccine administered opportunistically should be used in outbreaks involving drug users. PMID- 12889288 TI - Non-responders to hepatitis B vaccination: a review. AB - Hepatitis B virus (HBV) infection is a major public health issue throughout the world and vaccination of those at risk is the main method of containment. Of healthy vaccinees, 5-10% fail to mount an adequate antibody response. The antibody levels of an unknown further fraction of vaccinees fall considerably over time rendering them at a potential risk of infection. The scope of this article is to review the factors that might influence the immune response to HBV vaccination, to review the methods used to overcome the problem of poor response and to discuss what possible guidelines are available or needed in treating these vaccinees. PMID- 12889289 TI - Overlap between genitourinary medicine and public health in the surveillance and control of sexually transmitted infection in England, Wales and Northern Ireland. AB - The control of sexually transmitted infections (STI), and in particular outbreaks and other acute incidents of STI, often requires resources outside the genitourinary medicine (GUM) clinic. This study of Consultants in Communicable Disease Control (CsCDC) in England, Wales and Northern Ireland, explored the level of collaboration between district public health departments and GUM clinics and the usefulness at local level of STI surveillance data. The results show an inconsistent working relationship between GUM and public health throughout the country and highlight deficiencies in current surveillance systems. Participants in the survey suggested potential remedies to the problems identified including enhanced surveillance of STI and formalised relationships between GUM and local public health departments. Enactment of these suggested remedies may contribute to meeting the challenge of reversing current disease trends. PMID- 12889290 TI - Developing regional surveillance of sexually transmitted infection: a survey of the views of healthcare professionals in north west England. AB - Current surveillance of sexually transmitted infections in England and Wales relies on aggregated data collected by genitourinary medicine clinics. This system only allows limited analysis especially at a local level. A change to a disaggregate form of data collection could provide additional outputs at a local level. We undertook a survey of the views of those working in the prevention and control of sexually transmitted infections in the North West to determine what additional surveillance outputs would facilitate their work. Of respondents, 89% believed surveillance should deliver between four and nine types of analyses that can only be delivered by a surveillance system that is based on disaggregate data. Respondents indicated that it was acceptable for clinics to provide the data items required but that currently it was not feasible to do so because of workload constraints. PMID- 12889291 TI - Review of the use of neuraminidase inhibitors for prophylaxis of influenza. AB - Influenza causes significant morbidity and mortality particularly in vulnerable population groups. Although vaccination is recommended as the primary method of preventing influenza, antiviral drugs may play a complementary role. The National Institute for Clinical Effectiveness (NICE) has recently published guidance on the use of anti-viral drugs in the treatment of influenza, and further guidance on their use for prophylaxis is expected later in the year. In anticipation of this guidance this paper reviews the evidence for the effectiveness of the prophylactic use of neuraminidase inhibitors. Results of this review show that neuraminidase inhibitors are effective in preventing laboratory-confirmed clinical influenza in healthy adults, both when used as post-exposure prophylaxis for close and household contacts, and as seasonal prophylaxis in the wider community. Direct evidence in the elderly and institutions is too limited to draw firm conclusions about their effectiveness in that setting. However it seems probable that neuraminidase inhibitors would also be effective in these groups. PMID- 12889292 TI - Three years of low influenza activity--no reason for complacency. AB - Influenza activity in England and Wales has been unusually low over the last three years. Despite this, substantial morbidity and mortality has occurred in subgroups of the population. Furthermore, the influenza season 2001/02 was characterised by the emergence of a new subtype of the influenza A virus (H1N2), a timely reminder that it is not possible to assess the impact of a change in the virus strains circulating without having a comprehensive surveillance infrastructure in place. PMID- 12889293 TI - Delay in starting treatment for tuberculosis in east London. AB - In the United Kingdom there is little information about the delay between the onset of symptoms in patients with tuberculosis and the time it takes for them to be correctly diagnosed and treatment started. We have examined the duration and possible causes of such delay in our own district. The records of 93 patients were examined. Total delay in starting treatment was estimated as the time from the start of symptoms to commencement of chemotherapy. Patient delays were estimated from the time between the start of symptoms to the time taken to first attend their general practitioner (GP) with symptoms. Healthcare system delays were estimated from the interval between first being assessed by their GP and starting anti-tuberculosis treatment. Median total delay was 18 weeks (0-219). The time when patients first presented to their GP was determined for 64 patients: median patient delay was then estimated as nine weeks (range 0-104 weeks), and median healthcare delay five weeks, with a very wide range (0.5-210). Prolonged delay was seen in three patients with cervical lymph node disease. Patient delay was significantly longer than healthcare system delay (p = 0.019). Pulmonary disease was associated with shorter total delay in starting treatment compared with extra-pulmonary disease (p = 0.035). In patients with tuberculosis there were considerable delays in first presentation to medical services, in diagnosis and in starting treatment. Patient delays were longer than healthcare system delays. There is a need to improve awareness of the symptoms of tuberculosis both on the part of the general population and of health professionals, especially in areas of high incidence. PMID- 12889294 TI - Head lice diagnosed in general practice in the West Midlands between 1993 and 2000: a survey using the General Practice Research Database. AB - The potential of the General Practice Research Database (GPRD) for communicable disease surveillance was explored using head lice as an example. All diagnoses of head lice and prescriptions for parasiticidal agents from 1993 to 2000 in the West Midlands were analysed. Diagnoses reached a peak of 28.2 per 1,000 patient years at risk and total prescriptions reached a peak of 27.1 per 1,000 patient years at risk in 1997. Malathion and permethrin were prescribed most often. The proportion of further parasiticidal prescriptions issued within 30 days of the initial prescription increased to a peak of 11.5% of prescriptions in 1997. The ratio of the same:different further prescriptions changed during the study period, reaching a high of 5:1 in 2000. These trends are mirrored by the Royal College of General Practitioners (RCGP) Weekly Returns Service and Prescribing Analysis and Cost (PACT) data. Use of GPRD provides additional insights into patient data, particularly on prescribing, that would not be available from other sources. PMID- 12889295 TI - MMR vaccine uptake rates: a data validation study. AB - As part of our investigation into the decrease in the measles, mumps and rubella (MMR) vaccine uptake rates, we validated MMR vaccination records of all children born between 01/09/1998 and 31/08/1999 in our area (North Cheshire, South Cheshire, and Wirral). A significant number of children had received their MMR vaccine but were not recorded as such by the Child Health Computer System (CHCS). Reported COVER (cover of vaccination evaluated rapidly) data uptake (combined) for North Cheshire, South Cheshire, and Wirral Health Authorities for the period covered by the data validation study was 90.5%, the corrected uptake following the validation was 92.6%, 2.1% higher than the reported coverage. If the coverage data were to continue to form part of the NHS indicators of PCT performance, action by all PCTs to improve accuracy of immunisation data would be highly desirable. Electronic transfer of information from practices to the CHCS and between CHCSs, i.e. across boundaries, could improve data accuracy. PMID- 12889296 TI - Overcounting of black Africans in the UK: the problem of undetected record duplication. AB - Difficulties in recording unfamiliar African names, leading to inconsistent reporting of 'surname' code of the same individual, has raised concern that there is overcounting of newly diagnosed HIV-positive black Africans in the UK. Date of birth was used as a proxy indicator for duplication of entries of black Africans in the national HIV/AIDS patient database. Significantly more black Africans (59%) than whites (56%) share the same date of birth (p = 0.0023), and among black Africans certain birthdays occur at a very high frequency. Those born in Africa may not know their exact date of birth and so may be choosing, or have chosen for them, memorable or auspicious dates instead. After removal of individuals with birthdays consisting of the same number day and month, the evidence of disproportionate duplication of black Africans' dates of birth was weaker (p = 0.0129). Although this investigation provided evidence of selective failure to detect duplicated reports of the same individual among reports of HIV positive black Africans, in practical terms, the relatively small proportion (2.4%) of remaining excess duplicates is equivalent to 141 black African individuals on the database, and would not impact on the overall picture of the HIV epidemic in the UK. PMID- 12889297 TI - Outbreak of Legionnaires' disease on a cruise ship: lessons for international surveillance and control. AB - A sporadic case of Legionnaires' disease was linked to travel on a cruise ship. Investigation identified two further cases of Legionnaires' Disease and one case of non-pneumonic Legionella infection. An Incident Team confirmed the source to be the ship's water system and control measures were instituted that included pasteurisation, super chlorination and chlorine dioxide dosing. The Public Health Laboratory Service (PHLS) Communicable Disease Surveillance Centre (CDSC), through the European Surveillance Scheme for Travel Associated Legionnaires' Disease, identified three previous cases associated with the same ship's water system including one fatality. Lessons for the international surveillance and control of Legionnaires' disease on cruise ships are discussed. PMID- 12889298 TI - Meningitis C vaccine uptake by British undergraduates. AB - A new conjugated meningococcal group C vaccine became available in November 1999. This study set out to record the vaccination status of all first year undergraduates attending a university in South Wales in 2000-01 and to identify socio-demographic factors that might influence vaccine uptake. Vaccination status of 1,120 first year undergraduates (aged between 18-21 inclusive) was assessed by a questionnaire. Eight hundred and eighty-seven (79.1%) had been vaccinated prior to coming to university. Vaccinated undergraduates showed significantly more knowledge about meningitis than unvaccinated students. Both groups had only limited understanding. The vaccination rate was significantly higher (96.3%) among students offered vaccination in school compared to students not offered vaccination there (19.7%). This finding demonstrates the effectiveness of the school-based immunisation programme. Many of our sample did not understand that they had only been vaccinated for meningitis C. This, coupled with their poor understanding of meningitis symptoms, may result in students failing to respond appropriately to such symptoms. PMID- 12889299 TI - Urinary symptoms and Chlamydia. PMID- 12889300 TI - RNA interference pays off. PMID- 12889301 TI - Return to practice for healthcare scientists. PMID- 12889302 TI - Decontamination by fumigation. AB - Since the terrorist attack on the World Trade Centre on September 11th 2001, and the subsequent deliberate release of anthrax spores into the postal system of the USA, NHS hospitals and microbiology pathology laboratories have had to consider how they would respond to a release of a hazardous biological agent. This article looks at the procedure of fumigation of a known contaminated area following spillage or handling of a biological agent, and considers the additional problems that would occur should areas outside a laboratory, such as hospital wards or ambulances, require fumigation. PMID- 12889303 TI - Are performance targets overrated? PMID- 12889304 TI - Mind your back. PMID- 12889305 TI - Advising patients with diabetes about fasting during Ramadhan. AB - Fasting for Ramadhan is one of the five Pillars of Islam and is very important to Muslims. Nurses can help patients with diabetes to fast successfully by means of a pre-fast assessment, careful medicine management and good patient education. PMID- 12889306 TI - What you need to know about ... abdominal aortic aneurysm. PMID- 12889307 TI - Vaccine storage. PMID- 12889308 TI - Supplementary prescribing for the elective surgical patient. AB - Supplementary prescribing is an exciting new development in nursing. After qualifying, the supplementary nurse prescriber may prescribe a range of medications as part of a clinical management plan. The rationale for the choice of medication for inclusion in a clinical management plan for use in elective surgery is described. PMID- 12889309 TI - A cost-benefit analysis of training nurses for extended roles. AB - This study uses a questionnaire to assess the costs and benefits of a course to teach the extended roles of cannulation, venepuncture and intravenous drug administration. The course had a positive influence on nurses' uptake of venepuncture and IV drug administration, but not cannulation. The cost of 916 pounds to successfully train a single nurse in all three practices leads to the suggestion that the criteria for selecting nurses for the extended role are revised in order to achieve an improved cost-benefit ratio and methods for sustaining the newly acquired knowledge should be developed. PMID- 12889310 TI - University challenge. Interview by David Crouch. PMID- 12889311 TI - A fresh approach to osteoporosis. Interview by Carol Davis. PMID- 12889312 TI - Helping nurses to improve wound care. PMID- 12889313 TI - Support for troubled adolescents. PMID- 12889314 TI - Children with type 1 diabetes: Integrated Needs Assessment. PMID- 12889315 TI - Pseudo children: a cause for concern? PMID- 12889316 TI - Shared care: hospital, hospice, home. PMID- 12889317 TI - Aromatherapy and massage: the evidence. PMID- 12889318 TI - Undertaking research in practice. PMID- 12889319 TI - Complementary therapies: knowledge and attitudes of health professionals. AB - This cross-sectional, single-site project studied a random selection of qualified doctors and nurses in an NHS children's trust, using a combination of questionnaires and semi-structured interviews. The hypothesis tested was that: 'Health professionals at the trust neither feel able nor perceive a need to provide information about complementary therapies to families under their care'. Fifty questionnaires were distributed, 36 returned and 10 interviews completed. The study demonstrated complementary and alternative medicine (CAM) use was high among respondents and probably high among their patients. Attitudes were generally positive towards CAM use in children, although staff expressed concerns that CAM should do no harm. Knowledge of CAM was very limited but did not prevent staff from discussing CAM with patients or endorsing its use. It was felt CAM offered greatest benefit to children with chronic illnesses but that its most important role lay in the psychological support it could provide for both children and parents. PMID- 12889320 TI - Power and control over children and young people. AB - This feature introduces you to theories of control and power and invites you to relate these to your own practice with children and young people and the values of your work place. Rights-based approaches to care are outlined and through reading and Time Out activities, you will explore ways of challenging disempowering behaviours and make a personal action plan to empower children and young people. PMID- 12889321 TI - Sexual assaults during hostage takings and forcible confinements: implications for practice. AB - Hostage takings and forcible confinements are rare phenomena within our Canadian institutions. However, when they occur they cause enormous psychological and/or physical harm. A review of the literature revealed that no previous research on hostage takings or forcible confinements in a prison setting has been published. This investigation reviewed 33 hostage-takings/forcible-confinements spanning 11 years. The incidents were classified as follows: 20 hostage takings (3 with sexual assault) and 13 forcible confinements (7 with sexual assault). Sexual assaults were always against women and 36.6% of the women were sexually assaulted. This is a violent group of offenders with a prior history of sexual and nonsexual violence as well as escapes. While a quarter of the perpetrators had a forcible confinement or hostage taking in their current conviction, half had a prior history of such incidents. Rapists were overwhelmingly implicated in incidents that resulted in a sexual assault. Most perpetrators were below the age of 30 and serving sentences of less than 10 years in medium- or maximum-security institutions. The perpetrators of these incidents are not necessarily different from the general population of offenders in terms of their dynamic needs. Although this research is primarily descriptive in nature, it offers a unique contribution to the field by providing the first comprehensive description of this group of perpetrators. These findings are discussed in terms of their relevance to policy, operational practices, and the development of screening instruments. PMID- 12889322 TI - Distorted attitudes and perceptions, and their relationship with self-esteem and coping in child molesters. AB - Two studies were conducted to compare various aspects of child molesters, non sexual offenders, and community-based nonoffenders. These studies were aimed at examining some implications of our general model of sexual offending. Study 1 compared the responses of these 3 groups on measures of self-esteem, attitudes toward women and children, and social desirability tendencies. The only observed difference was that child molesters had significantly lower self-esteem than did the other 2 groups. In Study 2, child molesters displayed more cognitive distortions about sex between adults and children than did nonsexual offenders or nonoffenders. However in this study child molesters scored in the same range as the other participants on self-esteem and the tendency to use sex as a way of coping with problems. The results are discussed in terms of the differences between the present findings and earlier studies, and their implications for future research. PMID- 12889323 TI - A clinical model for the treatment of personality disordered sexual offenders: an example of theory knitting. AB - In this paper we present a clinical model for the treatment of personality disordered sexual offenders. The model is based on the theoretical assumption that cognitive distortions emerge from more general explanatory theories. It seems that the pathways of offending processes that are characterized by positive or negative affects can be found in all treated sex offenders regardless of their motivation. We assume that these pathways are activated by cognitive distortions that illustrate the underlying general explanatory theory held by the offender and that these distortions are typical for overall daily interpersonal behavior of the offender. This means that, as predicted by our clinical model in the case of offenders with a personality disorder, the type of explanatory cognition that is underlying their sexual offenses is the same as the one that is underlying their general explanatory theories. We will describe the clinical model and the treatment program used at Forensic Psychiatric Hospital Center Veldzicht with a review of our experiences. PMID- 12889324 TI - Adolescents' perceptions of the seriousness of sexual aggression: influence of gender, traditional attitudes, and self-reported experience. AB - Little is known about adolescents' perceptions of interpersonal aggression and the role of traditional social attitudes in these perceptions. Sixteen-year-old students (N = 212) of both sexes rated the seriousness of 9 aurally presented scenarios depicting either sexual or nonsexual, physical aggression. Sex of perpetrator and sex of victim were manipulated partly factorially. Students also reported on their own experiences (as perpetrators and victims) of the aggression portrayed, and completed a measure of traditional sex role ideology. Sexual aggression was rated as more serious than nonsexual physical aggression, especially when involving physical force. Girls gave higher seriousness ratings than did boys. Male-to-female aggression was rated as most serious, and male-to male aggression, least serious. Self-reported perpetrators tended to give lower ratings of seriousness than did victims. Traditional sex-role attitudes were associated with lower rated seriousness but not with reported perpetration or victimization. Multivariate analyses suggested that the effects of traditional sex-role attitudes could largely be subsumed by the effects of other study variables, especially participant sex. The role of attitudes as a direct cause of interpersonal aggression is discussed along with implications for intervention. PMID- 12889325 TI - Olfactory aversion and its mechanism of action represents an excellent summary of this useful technique. PMID- 12889326 TI - Bioethics in the United Kingdom: genetic screening, disability rights, and the erosion of trust. PMID- 12889327 TI - A view of bioethics from down under. PMID- 12889328 TI - Israel: bioethics in a Jewish-democratic state. PMID- 12889329 TI - Healthcare ethics in Finland. PMID- 12889330 TI - Biomedical ethics in Japan: the second stage. PMID- 12889331 TI - Changing roles in Russian healthcare. PMID- 12889332 TI - South Korea: human embryo research. PMID- 12889333 TI - End-of-life care in Turkey. PMID- 12889334 TI - CQ sources/bibliography. Bioethics now: international voices 2003. PMID- 12889335 TI - The ethical management of the noncompliant patient. PMID- 12889336 TI - Managed care, doctors, and patients: focusing on relationships, not rights. PMID- 12889337 TI - Response to "Reading futility: reflections on a bioethical concept" by Donald Joralemon (CQ vol 11, no 2). The rise and fall of death: the plateau of futility. PMID- 12889338 TI - Oregon v. Ashcroft: the battle over the soul of medicine. PMID- 12889339 TI - Hospice with a Zen twist: a talk with Zen hospice founder Frank Ostaseski. Interview by Steve Heilig. PMID- 12889340 TI - Age and economic asymmetries in the sexual relationships of adolescent girls in sub-Saharan Africa. AB - This literature review assesses the extent of age mixing and economic transactions in the sexual relationships of adolescent girls in sub-Saharan Africa and the behavioral dynamics of girls and men involved in these partnerships. The examination of more than 45 quantitative and qualitative studies finds that relationships with older partners and those that involve economic transactions are common and that these asymmetries are associated with unsafe sexual behaviors and increased risk of HIV infection. Although the reasons that adolescent girls engage in sexual relationships with older men are varied, receipt of financial benefits is a major motivation. The literature presents evidence that girls have considerable negotiating power over certain aspects of sexual relationships with older men, including partnership formation and continuation; however, they have little control over sexual practices within partnerships, including condom use and violence. The review discusses directions for further research and the implications of current knowledge for future interventions. PMID- 12889341 TI - Estimating induced abortion rates: a review. AB - Legal abortions are authorized medical procedures, and as such, they are or can be recorded at the health facility where they are performed. The incidence of illegal, often unsafe, induced abortion has to be estimated, however. In the literature, no fewer than eight methods have been used to estimate the frequency of induced abortion: the "illegal abortion provider survey," the "complications statistics" approach, the "mortality statistics" approach, self-reporting techniques, prospective studies, the "residual" method, anonymous third party reports, and experts' estimates. This article describes the methodological requirements of each of these methods and discusses their biases. Empirical records for each method are reviewed, with particular attention paid to the contexts in which the method has been employed successfully. Finally, the choice of an appropriate method of estimation is discussed, depending on the context in which it is to be applied and on the goal of the estimation effort. PMID- 12889343 TI - Cost-effectiveness of USAID's regional program for family planning in West Africa. AB - Between 1994 and 1996, the United States Agency for International Development (USAID) closed 23 country missions worldwide, of which eight were in West and Central Africa. To preserve United States support for family planning and reproductive health in four countries in that region, USAID created a subregional program through a consortium of US-based groups that hired mainly African managers and African organizations. This study assesses cost-effectiveness of the program through an interrupted time-series design spanning the 1990s and compares cost-effectiveness in four similar countries in which mission-based programs continued. Key indicators include costs, contraceptive prevalence rates, and imputed "women-years of protection." The study found that, taking into account all external financing for population and family planning, the USAID West Africa regional approach generated women-years of protection at one-third the cost of the mission-based programs. This regional approach delivered family planning assistance in West Africa cost-effectively, and the findings suggest that regional models may work well for many health and population services in small countries. PMID- 12889342 TI - Contraceptive use in a changing service environment: evidence from Indonesia during the economic crisis. AB - In the late 1990s, most Southeast Asian countries experienced substantial economic downturns that reduced social-sector spending and decreased individuals' spending power. Data from Indonesia were collected in 1997 (just before the crisis) and in 1998 (during the crisis) that are used in this study to examine changes in the contraceptive supply environment and in women's choices regarding contraceptive use. Despite substantial changes in providers' characteristics during the first year of the crisis, no statistically significant differences are found between 1997 and 1998 in overall levels of prevalence, in unmet need, or in method mix. Women's choices regarding source of contraceptive supplies, however, changed considerably over the period. Changes in the contraceptive supply environment are linked here to changes in women's choice of source of supply, and a number of providers' characteristics are found to be significantly associated with women's choices in this regard. PMID- 12889344 TI - Armenia 2000: results from the Demographic and Health Survey. PMID- 12889345 TI - Rwanda 2000: results from the Demographic and Health Survey. PMID- 12889346 TI - [Hyperparathyroidism: modern state of the problem]. AB - In the article, a review of the published literature is presented concerning the problem of primary and secondary hyperparathyroidism. Mechanisms are analyzed of secondary hyperparathyroidism in different pathological conditions. Improvements in methods for making diagnosis and an extensive application to practical use in clinical settings of such techniques as computerized tomography, bone densitometry, enzyme immunoassays to measure the parathyroid hormone, and other methods allowed the estimation of the prevalence of the condition under consideration and permitted raising a question as to a timely early diagnosis of hyperparathyroidism. In more than 50% of patients the above condition runs an asymptometic course being associated with the affection of the osseous system only, for which reason the earliest and most specific symptom diagnosable in hyperparathyroidism is considered to be osteoporosis. PMID- 12889347 TI - [Diagnostics and treatment of resistant arterial hypertension]. AB - Resistant arterial hypertension (RAH) is considered to be present in those cases where there is an increase in the arterial pressure (AP), if AP baseline level cannot be reduced down to 150/100 mm Hg and below that mark in conducting therapy with a triple-and-more drug regimens including diuretics and vasodilators prescribed in doses approaching the maximum ones, with the baseline (AP) level being 180/115 mm Hg and higher. The studies made permitted the development of a therapeutic RAH algorythm: use of inhibitors of the angiotensin-converting enzyme (ACE) in the complex therapy of RAH patients; naniprus is a drug of choice during therapy of low-renin refractory forms of AH; use of sodium nitroprussiate and inhibitors of ACE combined in treatment of RAH has been shown to ensure potentiation of a hypotensive effect of the above combination; prostenon is an effective means of correction of resistant forms of hypertensive disease; plasmapheresis is a reserve means in primary and symptomatic forms of RAH. PMID- 12889348 TI - [Pathogenetic mechanisms of nonsymptomatic myocardium ischemia]. AB - A review is submitted of relevant medical literature concerning the problem of silent myocardial ischemia in coronary patients. The role is described of coronary atherosclerosis, vasospasm, and disorders of hemostasis, its plasmatic link included, in the genesis of silent myocardial ischemia. PMID- 12889349 TI - [Bone metastasis in patients with breast cancer]. AB - Breast cancer (BC) is quite often accompanied by affection of the bones and alterations in mineral metabolism. The most important clinical manifestations of the above processes that appeared to a considerable extent to be a cause of a poor condition of patients include pains in bones, pathological fracture and hypercalcemia. Metastatic events in the skeleton bone are recordable in 13.5 to 85 percent of cases, as evidenced by autopsy findings. In BC patients presenting with hormone-dependent tumours and metastases in the bones, indices for survival tend to be much higher than in those patients with solid tumours of other localization. The higher level of survival in such patients warrants an effective palliative treatment to be instituted to improve quality of their lives. PMID- 12889350 TI - [Enhancing the efficiency of sanatorium and resort rehabilitation of the liquidators of the Chernobyl accident by hyperbaric oxygenation]. AB - Results of our studies permit reaching the conclusion that oral intake of the mineral water Naftusya as the leading factor with an additional prescription of the course of the hyperbaric oxygenation procedures promote enhancement of functional reserves, stabilize free-radical processes and bodily system of antioxidant defence, favour stabilization of cell membranes, initiate decorporative action, diminish depression of immunity and supply oxygen to those enzymic systems whose activity has been adversely affected by hypoxia. PMID- 12889351 TI - [Cardiovascular disorders in patients with pheochromocytoma]. AB - The present study focuses on variants of cardiovascular disturbances in patients with pheochromocytoma, on identification of certain pathogenetic aspects of disturbances of the heart's rhythm, on the study of the impact of surgical treatment on hemodynamic shifts. A retrospective analysis was performed of 100 clinical records of those patients having had surgery for pheochromocytoma over the last five years. It has been ascertained that a major proportion of tumours of the chromaffin tissue of the adrenals secret epinephrine and norepinephrine giving rise to a mixed form of arterial hypertension. In 61 percent of patients, arterial hypertension tended to be accompanied by hypertrophy of the left ventricle and its systolic overload (3%) that has been noted to dispel after surgical treatment. In 27 percent of patients with pheochromocytoma, against the background of hypertensive crisis, abnormalities of metabolism or unstable coronary circulation there come to be seen paroxysms of disturbances of the heart's rhythm (extrasystole, paroxismal tachycardia) and dysfunction of the sinoatrial node. PMID- 12889352 TI - [Cellular and humoral immunity in patients with acute disorder of coronary circulation]. AB - The condition was studied of cell-bound and humoral immunity in patients with ischemic heart disease. The data secured permit reaching the conclusion that the unstable clinical course of angina pectoris is accompanied by unspecific alterations in the cellular link of immunity such as increase in the absolute number of T-lymphocytes and T-suppressors. In patients with myocardial infarction, in the acute period of the illness there develop inflammatory reactions involving participation of cell-bound (neutrophilic leucocytosis, a drop in the absolute number of acidophilic granulocytes, absolute number of lymphocytes, T-lymphocytes, T-suppressors, increase in T-helpers--T-suppressors ratio; decline in T-lymphocytes activity) and humoral (a drop in the level of Ig A, increase in blood serum concentration of circulating immune complexes) links of immunity. PMID- 12889353 TI - [Malonic dialdehyde level in the blood and lungs in complicated course of acute myocardial infarction]. AB - Studies were conducted to measure the content of malonic dialdehyde in the venous and arterial blood and in tissues of the lungs in uncomplicated and complicated courses of acute myocardial infarction (MI). It has been ascertained that in the development of MI complications there is noted an interrelationship of the degree of intensification of free radical processes in the blood and tissues of the lungs, which observation is of pathogenetic significance. PMID- 12889354 TI - [Efficacy and safety of treatment with methotrexate, leflunomide, detralex, and their combination of patients with rheumatoid arthritis]. AB - A comparative evaluation was done of efficacy and safety of methotrexate, leflunomide singly and of combination of methotrexate with leflunomide or detralex. A total of 189 patients with rheumatoid arthritis (RA) were examined. A 6-month course of controllable treatment was instituted in them. The time-related course of clinical-and-laboratory indices allowed judgement about efficiency of the treatments administered. The functional condition of the patients was assessed according to HAQ. As to efficacy and toxicity, leflunomide (in a dose of 20 mg/daily) was comparable to methotrexate (7.5 to 10 mg/per week) whereas the combination leflunomide-methotrexate has been shown to considerably accelerate regression of clinical symptoms of RA while the use of detralex in the therapeutic complex proved to enhance efficiency of pharmacotherapy with methotrexate and to reduce the incidence rate of its side effects. PMID- 12889355 TI - [Autonomic control and functional condition of suprasegmental structures of the brain in patients with heart rhythm disorders and vasculo-autonomic dystonia]. AB - A study was made by the method of combination of electroencephalography (EEG) and variational pulsimetry in 157 patients to determine the background vegetative tone and that very tone during conducting functional tests aimed at activating of sympathetic and parasympathetic portions of the nervous system. Criteria have been established characterizing the vegetative tone. Particular features are described of changes in the power of the EEG wave spectra while conducting tests in those groups being different in their baseline vegetative tone. The percentage is estimated of the incidence rate of the cardiac rhythm disturbances in those groups being different in their vegetative tone. PMID- 12889356 TI - [Albumin differentiation by isoelectric focusing in patients with liver cirrhosis]. AB - In patients with cirrhosis of the liver, there is noted a striking imbalance in the distribution of isoelectric fractions of serum albumin in the isoelectric field. Disproportion in isoelectric fractions of serum albumin can, we believe, be referred to functional loads on albumin relative to fixation and transport of surplus of bilirubin and ammonia. In the ascitic fluid, there comes to be seen, in fact, a reversal of imbalance of pH-dependent fractions of albumin,--probably, due to "purification" in the lymphatic system and partly in the functioning hepatocytes. As to the isoelectric spectrum, the ascitic fluid albumin is qualitatively very similar to serum albumin of healthy people. PMID- 12889357 TI - [Endoscopic characteristics of chronic bronchitis treated with substitution of surfactant]. AB - An analysis has been performed of endoscopic management of chronic bronchitis with making use of substitution surfactant therapy with a home-produced drug preparation of the exogenous surfactant sucrim. Positive results obtained in the studies conducted permit the substantiation of a 2-fold use of the above drug in a combined treatment. Visual normalization of bronchial mucosa in purulent diffuse endobronchitis, degree III intensity of inflammation, was seen by day 8 to 12 from the commencement of therapy. PMID- 12889358 TI - [Diagnostic and prognostic ability of the peripheral mononuclear lymphocyte to produce nitric oxide in multiple sclerosis]. AB - The article presents data on the chief mechanisms of human cell apoptosis, on the role of nitric oxide (NO) in modulation of the above mechanisms in cells of normal tissues and organs and in multiple sclerosis (MS) as well. Aspects are analyzed of NO action in the processes of apoptosis in MS. Corticosteroid pulse therapy is recognized as a powerful inducer of T-lymphocyte apoptosis in the condition under consideration. PMID- 12889359 TI - [Immunogenetic characteristics of nuclear families of adolescents with delayed sexual development]. AB - The paper submits data on the antigenic composition of HLA system in members of nuclear families of adolescent boys presenting with the delay in sexual maturation. Positive and negative associations of particular antigens with sexual underdevelopment are stated. Estimated in the families was the highest incidence rate of gaplotypes A 28-B 40, A 28-B 8, A 1-B 40, and A 3-B 40. The authors come out with a suggestion that it might be possible to use the identified antigens in prognostication of delayed puberty. PMID- 12889360 TI - [Clinical criteria of the risk of oncological diseases in women as a basis for prevention of reproductive system diseases]. AB - A multifactorial analysis was done of clinical past history of 520 female patients with cancer of the ovary, 482 patients with cancer of the endometrium, and of 513 essentially healthy women (control group). 23 historical and clinical sings were regarded as risk factors for development of cancer of the ovary and endometrium (disordered menstrual and reproductive functions, chronic inflammatory processes, benign tumours of the uterus and adnexa uteri, pretumour affections of the skin, and other pathologies). Of these, 15 sings were recognized as being of high informative value. PMID- 12889361 TI - [Antithrombin III level in patients with acute viral myocarditis]. AB - The article focuses on the status of a natural anticoagulant antithrombin III in patients with acute viral myocarditis (AVM) and on modes of treatment thereof. It has been proved that there is a statistically significant correlation between a drop in concentration of antithrombin III and degree of severity of AVM. Convincing, statistically significant data have been obtained that antithrombin III gets increased in AVM patients undergoing complex therapy: in those patients running a mild course of the illness, a mild one presenting with elevated indices for homeostasis, a medium gravity course (diclofenac, heparin, thiotriasoline, quick-frozen plasma), and grave course as well (prednizolon, heparin, thiotriasoline, quick-frozen plasma), which fact can be taken account of in choosing a therapeutic regimen. PMID- 12889362 TI - [Blood peroxidase activity in patients with cerebral atherosclerosis receiving microwave resonance therapy]. AB - Results are submitted of studies designed to measure peroxidase activity in the blood of 41 patient presenting with atherosclerosis-associated cerebrovascular disorders, while receiving microwave resonance therapy. Those patients presenting with cerebrovascular pathology proved to have the peroxidation syndrome as was evidenced by the analysis of the studies made. The above syndrome can be reversed with the aid of the microwave resonance therapy. PMID- 12889363 TI - [Cholesecretory function of the liver and correction of its disorders in patients with cholesterosis of the gallbladder]. AB - The article presents data on the state of the cholesecretory function of the liver and on correction of its disturbances in patients with cholesterosis of the gallbladder. The use of lecitin and ursofalc against the background of the ration energy value-restricted, food fibre-increased differentiated diet favours augmentation of the bile content of bile acids, normalization of the cholatho cholesterin ratio. PMID- 12889364 TI - [Treatment of patients with acute pneumonia]. AB - Efficiency was studied of treatment of acute pneumonia (AP) in 148 patients with a focal croupous form. In the grave course of AP, antibacterial therapy with benzilpenicillin combined with sulphanilamide agents biseptol and nitrofuran was tried, the correcting therapy having been instituted on obtaining findings from assays of the sputum for sensitivity of the microflora to antibiotics. In those cases with Friendlander's bacilli recoverable, methicillin, oxacillin, macrolids, cephalosporins and some other agents were antibiotics of choice. Administered in the atypical course that was related more frequently to mycoplasms or chlamidia, was erythromycin phosphate i.v., lincomycin. In Staphylococcus-associated pneumonia, there have been used antibiotics resistant to penicillinases and endowed with an inhibitory activity toward staphylococci--oxacillin, methicillin, cephalosporins, chlorophenicols or chlorophphyllipt i.v. combined with an antistaphylacoccal plasma, antistaphylococcal gammaglobulin or hyperimmune plasma. The following drugs were prescribed to ensure bronchial patency and liquefaction of the sputum--euphyllin, mucalthin, bromhexin, lasolvan that are known to stimulate production of sulfoctant. The therapeutic complex comprized fibs, aloe, apilac, prodigiosan, solcoseryl, methyluracil known to stimulate bodily reactivity and resolution of inflammatory infiltration, nicotinic acid, heparin that have been shown to improve micorcirculation, tocopherol, unithiol known to regulate lipid oxidation. The data secured suggest to us a sufficient efficacy of the above-outlined therapy of AP. PMID- 12889365 TI - [Side effects of chemotherapy in patients with pulmonary tuberculosis and concomitant diseases]. AB - As many as 395 patients with pulmonary tuberculosis concurrent with other illnesses were examined. It has been ascertained that in 57.8 percent of cases there occurs a side effect in the wake of the antituberculous drug administration. The following were recognized as ill effects of the drugs employed--allergic reactions, candidiasis, disbacteriosis, toxic reactions,--in 25.44%, 3.07%, 7.02%, and 64.47% of cases respectively. Revealed in the above series was a marked side effect on the digestive organs, neurotoxic reactions, toxic effect on the cardiovascular system in 28.08%, 15.35%, 10.53% of cases respectively. The most frequently seen side effect was due to rifampicin (23.25%), streptomycin (18.42%), isoniazid (17.08%). The least number of complications was with taking etambutol--3.94%. In 43.42% of cases, the identified side effect necessitates withdrawal of the drug, in 56.58% of cases, the management of the concomitant medical problems, a change in routes of administration of medicines, and combined employment of antihistaminic drugs and hyposensitizing methodological approaches to the management of the condition permitted carrying on with the therapy instituted. PMID- 12889366 TI - [Application of pneumoperitoneum in collapse surgical procedures during tuberculosis outbreak]. AB - Under conditions of tuberculosis epidemic resective methods come to be used more and more seldom, the causes of which fact being resistance of Koch's bacilli to antibacterial drugs, extension and duration of the process. Collapsosurgical interventions constitute an alternative to resecting methods but such operations are accompanied by complications in 11 to 22 percent of cases. The author suggests that pneumoperitoneum be used to prevent complications developing in thoracoplasty, substantiating his suggestion, validating it by submitting the relevant statistical material. PMID- 12889367 TI - [Treatment of disorders in interferon status of patients with urogenital chlamydial infection]. AB - The interferon status was studied in 54 patients with urogenital chlamydiosis, presenting with disturbances in the reproductive system. The blood serum content of gamma-interferon was 32-fold in the above patient population vs control subjects on conventional therapy. Supplementing such a therapy with cycloferon, an inducer of endogenous interferon, and with immunomodulin have been shown to activate production of interferon. PMID- 12889368 TI - [Diagnostics and treatment of sleep disorders in elderly people]. AB - A meta-analysis was done of the published literature concerning the modern status of the issue of sleep disturbances and of findings from the author's investigations. Data are submitted obtained in a clinical examination of 61 elderly patient in a therapeutical hospital. In the above patients, high prevalence has been revealed of different sleep disturbances. Insomnia is often a sequela of psychophysiological derangements, which fact necessitates an exhaustive differential diagnosis and choice of remedies to be used in therapy of the above disorders. An experience is described of employment of zopiclon (Somnol Grindex) to deal with sleep problems in those patients in the involutional period. It is suggested that psychiatrist services be more actively used to give the necessary advice to those patients of the cardiological profile presenting with sleep disturbances. PMID- 12889369 TI - [Species affiliation of staphylococci isolated in purulent infections]. AB - The microflora was studied of the clinical material taken from those patients presenting with different pyo-septic diseases. It has been ascertained that a key role in the etiology of the above diseases must be referred to gramme-positive micro-organisms--to coagulase-negative staphylococci among their number. Of these, S. haemolyticus and S. epidermidis were found with high frequency. PMID- 12889370 TI - [Diagnostic significance of tests with toxoplasmin in clinical practice]. AB - Intracutaneous test (ICT) with toxoplasmin is a technically simple and inexpensive method of laboratory diagnosis of toxoplasmosis. Of the 5472 examined sick persons, 54.28 percent displayed a positive ICT. With a positive ICT, any titer of any serological reaction of toxoplasmosis is regarded as a diagnostic titer. PMID- 12889371 TI - [Remote results of the treatment for stage I-II breast cancer]. AB - A retrospective analysis was carried out of an 8-year survival in a series of 410 patients presenting with stage I-II breast cancer. Results are assessed of different combinations of a multiple-modality treatment involving surgical, chemotherapeutical, radiation, and hormonal methods. It is pointed out that adjuvant chemotherapy can make the results better. Other methods do not exert statistically significant effect on survival of patients. PMID- 12889372 TI - [Radiation therapy in treatment of the sclerotic form of pathological rearrangement of bone tissue of the lower extremities]. AB - An analysis was done of treatment of 77 servicemen with a sclerotic form of pathological rearrangement of bones of the inferior limbs. Used in the above subjects was oteoscintillography that promoted the accuracy of diagnosis and allowed the study of the time-related course of the process in bones, which measure permitted correcting the therapeutic regimen designed to deal with the pathology under consideration. The complex of conservative treatment was supplemented by a course of gamma-radiation therapy in a cumulative dose of 1.6 Gy, which measure has been shown to yield better results as compared with the control group of patients. PMID- 12889373 TI - [Modern views on the theory of nature (mizadj) by Ibn Sina in medicine]. AB - On the basis of the theory of mizadj Ibn Sina advocates an individual approach to a person, regardless whether that person might be healthy or sick, infant or advanced in years, with taking account of a unique combination of all his/her inherences and concrete conditions of his/her living and activities, starting with which principles Ibn Sina suggests criteria for defining the notion "mizadj", each such criteria corresponding to a certain division in relevant lines of scientific inquiry. The teaching of Ibn Sina about mizadj at the present stage assumes ever greater importance in medicine both in diagnosis and treatment and in prophylaxis against diseases, which facts attest to the need for a thoroughgoing approach to the study of creative legacy of a great scientist. PMID- 12889374 TI - [Morphological characteristics of spermatogenesis in experimental juvenile hypothyroidism in rats]. AB - Partisular features have been studied of spermatozoa and cells of spermatogenesis in rats in experimental hypothyroidism in the prepubertal period. Common features to be seen in the wake of hypothyroidism include a decline in the locomotor activity of spermatozoa and abnormalities in maturation of cells of spermatogenesis, which facts necessitate restoration of the thyroid function to prevent disorders of spermatogenesis in the fully-developed organism. PMID- 12889375 TI - [Effect of phlogenzym in long-term treatment of patients with multiple sclerosis]. AB - An assessment was carried out of clinical effectiveness of the drug phlogenzym in 74 patients with remitting, remitting-progressive, and secondary progressive course of multiple sclerosis. Phlogenzym intake for up to one to three years resulted in decline in the incidence of complications, with their degree having come to be lower, duration of remissions longer, progression of the illness slowed down. The data secured suggest to us that phlogenzym is a safe agent. It can, we believe used in a therapeutic regimen for those patients presenting with remitting and remitting-progressive types of the course of the disease. PMID- 12889376 TI - [Effect of erbisol on functional activity of T-helpers of type II in production of IL-4 and IL-10 in vitro]. AB - Studies were conducted on effects of different doses of the drug preparations erbisol and super erbisol on the functional activity of type II helpers T lymphocytes in healthy subjects in respect of in vitro production of interleukine 4 (IL-4) and interleukine-10 (IL-10). The data secured permit regarding the above drugs as immunomodulating agents promoting reduction of secretion of such important mediators of the immune system as IL-4 and IL-10. PMID- 12889378 TI - [Biological age of the workers of plants producing thermo-, sound-, and asbestos isolating materials]. AB - Determination of biological age in workmen helps towards the carrying out of an analysis of a contribution that factors of the working environment and labour process make into age-associated decline in vital capacity, screening for detection of those persons bearing a high risk for a disease that might be contracted/sustained in the course of the employment; it can be used in early diagnosis and in implementing better measures designed to administer primary and secondary prophylaxis of diseases arising out of the employment, due to health hazard exposure in industrial settings, and of occupational diseases. The findings from the conducted studies on biological age of workers of the thermal and-sound plants and asbestos product plants suggest to us a growing rate of aging. PMID- 12889377 TI - [Effect of apisatron in lumbar spinal cord diseases]. AB - A therapeutic effect has been studied of the drug apizartron of the ESPARMA GmbH firm (Osterweddingen, Germany) in those patients presenting with damaged lumbar spine. The ointment apizartron has been shown to alleviate the pain syndrome and myotonic (spastic) reactions, with its effect having been somewhat less pronounced on radicular compression, neurodystrophic disorders, and on the arterial bloodflow of the inferior limbs. The drug is recommended for use in a complex therapy of vertebrogenic disorders of the lumbar spine. PMID- 12889379 TI - [Evaluation of vital activity of workers with obliterating diseases of lower extremities servicing electric transmission lines]. AB - In the transmission lines (750 kw, 50 Hz) workers, the functional condition of arteries of the inferior limbs was evaluated, with the pathogenetic relation having been substantiated of adverse factors of the electromagnetic field with development of thromboobliterating diseases of arteries of the lower extremities, the rehabilitation potential established together with criteria for expert evaluation of the work fitness. PMID- 12889380 TI - [On the "medical institution regulations"]. AB - Stated in the article on the basis of the author's personal experience and summarized materials of the study of the "Medical Institution Regulations" are main principles and sections of this document that is the chief state-paper regulating activities of a medical institution. The importance is shown of the "Regulations" for a medical institution to function properly under new social-and economic conditions. PMID- 12889381 TI - [Marketing aspects of managing of health care delivery to patients with sexually transmitted infections in modern practice]. AB - The article is devoted to the study of scientific-and-practical researches carried out in the field of management, organization, and marketing of medical care provided to patients presenting with infections transmitted through sexual contacts. The conducted investigations proved to be of current concern as evidenced by the analysis of the relevant literature now available, based on which analysis major lines of further research have been outlined, that include problems of marketing, process, strategic management and planning of activities of the health system in the face of the impact of the external conditions and within the framework of the market economy existing in the country. PMID- 12889382 TI - African nurses targeted in HIV drug project. PMID- 12889383 TI - A continent under siege. PMID- 12889384 TI - Nurses speak out on AIDS. PMID- 12889385 TI - Made to measure. Interview by Kay Smith. PMID- 12889386 TI - Primary concern. Interview by Elizabeth Lynch. PMID- 12889388 TI - Put the records straight. PMID- 12889389 TI - On speaking terms. PMID- 12889390 TI - Teenage kicks. PMID- 12889391 TI - School's out. PMID- 12889392 TI - An essential care course for healthcare assistants. AB - There is growing consensus that healthcare assistants (HCAs) need access to professional training, education and support. The authors describe a short course designed to educate and support HCAs, focusing on the care of older people. PMID- 12889393 TI - The peripherally inserted central catheter. AB - Peripherally inserted central catheters (PICCs) are commonly inserted by nurses. This article gives an overview of the care and management of the PICC in general adult nursing. PMID- 12889394 TI - Post-operative wound infections. AB - AIMS: To examine the factors influencing post-operative wound infections, such as the patient's age, sex, type and duration of surgical procedure, length of hospital stay, and the type of antibiotic prophylaxis used. In addition, to examine the level of nurses' and physicians' knowledge and application of aseptic technique procedures pre-operatively, peri-operatively and post-operatively. METHOD: The study was conducted between 1997 and 1999 at two major hospitals in Greece. One hundred and twenty six (n = 126) patients and 90 (n = 90) healthcare professionals (74 nurses and 16 physicians) were included in the study. Healthcare participants were interviewed and completed questionnaires that were designed to measure demographics, experience, infection control training and education, staffing levels, perceived deficits, and managerial support. RESULTS: The study showed that there was a high rate of infection (17.5 per cent) in both hospitals with more predominance in the male population (p < 0.05). The length of hospital stay and type of surgical procedure were found to be significant factors in infection. In addition, the lack of awareness of aseptic techniques among nurses and physicians in several practice areas, despite the availability of a large number of relevant and well-established research findings and guidance, was also found to be significant. The duration of the operation was found to be correlated with infection (R = 0.208, p < 0.01). Emergency surgical procedures were more susceptible to infection (p = 0.08) than scheduled procedures. The results will provide hospital administrators with strategic goals and actions that might have a significant impact on reducing infection outbreaks following surgery. CONCLUSION: Continuous education of all healthcare professionals in basic infection control procedures, policies and new information on antimicrobial resistance is recommended to increase detection and identification of surgical wound infections. A follow-up study should be conducted to measure the improvement in the use of aseptic techniques by both nurses and physicians. PMID- 12889396 TI - Telemedicine in educational settings. AB - The use of telemedicine to enhance patient care is well documented in the literature (Currell et al 2001); however, its use in educational settings requires further exploration. Technological advances in electronic communication have been the catalyst for enabling the transmission and storage of large volumes of data. This, in turn, has allowed still and video images to be used for clinical consultation and the advancement of healthcare professionals' knowledge and skills. This article discusses the use of telemedicine in healthcare practices and explores its value as an educational tool, particularly in the field of wound care. PMID- 12889397 TI - Intermittent pneumatic compression pump settings for the optimum reduction of oedema. AB - This study investigated the reduction in lower limb oedema achieved when different combinations of intermittent pneumatic compression (IPC) pump settings were used. Twenty-seven individuals aged between 22 and 96 years, with ankle oedema of venous origin, were randomised to either a treatment or control group. Treatment was delivered via a Flowpac pump (Huntleigh Healthcare Ltd, Luton, UK) possessing variable controls. The control group were 'treated' by elevating both lower limbs. The average difference in limb volumes was compared, for each combination of pump setting, to changes in limb volume in the control group. These comparisons revealed that the highest mean reduction in limb volumes was recorded for a pressure of 40 mmHg (p = 0.02), 10 second deflation time (p = 0.0002) and 15 second inflation time (p = 0.0096). Other significant results were obtained by the 30 mmHg pressure (p = 0.017), 35 second deflation time (p = 0.018), and 5 and 45 second inflation times (p = 0.012, 0.013 respectively). For many setting combinations no significant differences were observed between the limb oedema seen in the control and experimental groups. The results should be interpreted with caution because of the large number of tests performed. In addition adverse effects were observed in six out of nine subjects at 70 mmHg pressure and this pressure was discontinued. One subject suffered cramp at 60 mmHg pressure. Lower pressures together with shorter inflation and deflation times appear to be more efficient than higher pressures and long inflation/deflation times. In addition the lack of significant reduction in oedema at pressures above 40 mmHg suggests that the higher pressures cause a tourniquet effect. PMID- 12889395 TI - Pressure ulcer risk: audit findings. AB - This article describes the findings of a pilot audit project that assessed the effects on practice and patient care of guideline recommendations for reducing pressure ulcer development. A national audit project to enable the implementation strategy and audit tools to be evaluated across a range of healthcare settings and patient groups should now be undertaken. PMID- 12889398 TI - Randomised controlled trial to evaluate a new double-layer air-cell overlay for elderly patients requiring head elevation. AB - A clinical investigation was conducted concerning the effects of a newly designed double-layer air-cell overlay in preventing the onset of pressure ulcers for patients with a Braden scale score of < or = 16, and who require a head-elevated position of 45 degrees or higher. A randomised controlled trial was undertaken involving 82 patients from a general hospital ward using one of the following three support surfaces: a double-layer air-cell overlay, a single-layer air-cell overlay or a standard hospital mattress. A significantly lower percentage of patients using the double-layer air-cell overlay developed pressure ulcers (3.4%) compared to 19.2% and 37.0% for those patients using the single-layer air-cell overlay and standard mattress respectively. Based on these findings, a double layer air-cell overlay should be more effective in preventing the onset of pressure ulcers than either a single-layer air-cell overlay or a standard hospital mattress for subjects requiring head elevation. PMID- 12889399 TI - The effect of a specialist seating assessment clinic on the skin management of individuals with spinal cord injury. AB - OBJECTIVE: To evaluate the specialist seating clinic's effectiveness in improving skin management knowledge and independence, represented by the Needs Assessment Checklist (NAC). DESIGN: Longitudinal, between subjects design, with two intervention groups and one control. SETTING: Tertiary care, spinal cord injury centre (National Spinal Injuries Centre), Stoke Mandeville Hospital, United Kingdom. METHOD: This study assessed the skin management ability of three groups. Group 1 consisted of individuals who had attended a specialist seating assessment (SSA) clinic before their first needs assessment, group 2 had attended SSA between their first and second needs assessment, and group 3 (control) had not attended at all. Patient skin management ability was assessed using the skin management subscale of the NAC, a measure of rehabilitation outcome, at two time points. RESULTS: Significant differences were identified between group 3 and group 1 at both the first (t = 2.36, degrees of freedom (df) = 37, p < 0.05) and second (t = 2.84, df = 37, p < 0.01) needs assessment. Significant improvements were also observed within each group between the first and second needs assessment time points in all seating assessment categories. CONCLUSION: Skin management achievement scores were significantly higher for patients who had attended a SSA clinic before their first NAC (group 1) at both time points, supporting the use of SSA as a proactive intervention to improve patient independence, knowledge and awareness, and potentially reduce pressure ulcer incidence. PMID- 12889400 TI - Integrated care pathway: the prevention and management of pressure ulcers. AB - The introduction of this pathway for the prevention and management of pressure ulcers was influenced by the development of the pathway for the management of a patient with a fractured neck of femur. Systematic documentation of preventative measures for patients with a fractured neck of femur helped reduce the incidence of pressure ulcers in this high-risk group. The original draft was informed by evidence-based clinical practice guidelines. This draft of the pathway was piloted, reviewed and changed in line with current evidence and comments from users. It was invaluable to pilot the pathway and review as necessary in order to make this a true workable document that would enable continuity of care, help prevent hospital-acquired pressure ulcers and be an effective teaching aid. A trust-wide retrospective audit of documentation for the prevention and management of pressure ulcers has been conducted, which found that the pathway facilitated a more concise approach to inform care than standard documentation. A further audit has been undertaken in the care of the elderly directorate which has demonstrated various ways in which the pathway had been completed. Challenges remain for the effective use of this documentation. Further education needs to be completed in order to promote multiprofessional use of the pathway. The pathway is now being integrated into others, which will increase awareness and involvement of other professional groups in this important aspect of patient care. Ongoing audit will take place to assess the impact upon patient outcomes. PMID- 12889401 TI - On the road to joining the faculty. PMID- 12889402 TI - Dress rehearsal. PMID- 12889403 TI - Tackling injury head on. PMID- 12889404 TI - Auditing a nurse led fracture referral service. PMID- 12889405 TI - Chill out: therapeutic hypothermia improves survival. PMID- 12889406 TI - Ankle sprain: how much do A&E staff know? PMID- 12889407 TI - The Access to Medical Treatment Act: reclaiming America's health care freedom. PMID- 12889408 TI - AHNA is A-buzz with the news about education. PMID- 12889409 TI - Integrative reflexology. PMID- 12889410 TI - Holistic pain management. PMID- 12889411 TI - Plastics: everything you need to know to improve your health and the environment. PMID- 12889412 TI - Introducing the Pranic Healing system to holistic nursing practice. PMID- 12889413 TI - The view from Carabelli's cusp. Strategy vs. tactics. PMID- 12889414 TI - Spending for dental care: a positive outlook even in a recession. PMID- 12889415 TI - Perfectionism. PMID- 12889416 TI - Ethical dilemma. PMID- 12889417 TI - Oral and maxillofacial pathology case of the month. Odontogenic myxoma. PMID- 12889418 TI - After hours care. PMID- 12889419 TI - Reading people. PMID- 12889420 TI - The migraine test. Would you be "wrong" or "fooled"? PMID- 12889421 TI - Hot on the inside. AB - When a disease process becomes life-threatening, it is termed to be malignant. Hyperthermia is a heat illness that arises from one of two basic causes: 1) the body's normal thermoregulatory mechanisms are overwhelmed by the environment (an exogenous heat load) or, more commonly, by excessive exercise in a moderate-to extreme environment (an endogenous heat load); or 2) failure of the thermoregulatory mechanisms, such as those encountered in the elderly or debilitated patient. Either cause can lead to heat illnesses such as heat cramps, heat exhaustion or heatstroke. Heat cramps are brief, intermittent and often severe muscular cramps that frequently occur in muscles fatigued by heavy work or exercise. They are believed to be caused by a rapid change in the extracellular fluid osmolarity resulting from sodium and water loss. Heat exhaustion is a more severe form of heat illness characterized by minor changes in mental status (poor judgment, irritability), dizziness, nausea and headache. In severe cases, the patient may have an altered LOC. Just as with heat cramps, profuse sweating is present. Removing the patient from the hot environment and administering fluids will usually result in a rapid recovery. [table: see text] Left untreated, heat exhaustion may progress to heatstroke. Heatstroke results when there is a complete collapse of thermoregulatory mechanisms. This will lead to a rise in body core temperature in excess of 105.8 degrees F (41 degrees C), which will produce multisystem tissue damage and physiological collapse. Severe cases can cause death. The patient in this case had an axillary temperature taken and recorded at 101.4 degrees F. Typically, axillary temperatures are one degree cooler than oral temperatures, which are one degree cooler than core temperatures. This patient, then, had a core temperature of 103 degrees F or higher. There are two types of heatstroke: classic and exertional. Classic heatstroke occurs during periods of sustained high ambient temperatures and humidity. Exertional heatstroke more often occurs in athletes, military personnel and people who work strenuosly in the environment. In these situations, endogenous heat accumulates more rapidly than the body can dissipate it in the environment. Although sweating is usually absent in the classic form of heatstroke, 50% of exertional heatstroke cases have persistent sweating as a result of catecholamine release. The presence of sweating does not preclude the diagnosis of heatstroke, and cessation of sweating is not the cause of it. As the illness progresses, peripheral vasodilation occurs, resulting in hypotension and shunting. As internal temperatures rise, myocardial contractility begins to decrease, manifested by bradycardia and irritability of the myocardium. No matter the age group, the presence of hypotension and decreased cardiac output indicates a poor prognosis for the patient. PMID- 12889422 TI - First impressions count. PMID- 12889423 TI - Dealing with your enemies. PMID- 12889424 TI - Motor vehicle accidents. PMID- 12889425 TI - Commotio cordis. When sudden death isn't a sporting term. PMID- 12889426 TI - Deadly defects. Managing the pediatric patient with a congenital heart defect. AB - CHD-specific emergencies may be managed quite effectively in the prehospital environment. The key to successful prehospital management of CHDs is identifying the cardiac anomaly, obtaining an effective history and physical assessment, and providing supportive care. When obtaining a history, it is important to remember that parents know their children and their children's diseases very well. An EMS provider who ignores information from a parent is doomed to failure. Realizing that most EMS providers are not well-versed in the various CHDs and are not likely to have a great deal of experience in working with CHD patients, conferring with medical control prior to initiation of any treatments is strongly encouraged. Finally, remember that patients with CHDs are sick and have traditionally been sick for quite a while. Because of the chronic illness, it is easy to focus on the congenital defect when called to the scene for an ill child and neglect the potential for a noncardiac-related pathology. Although the most commonly seen congenital heart defects have been reviewed here, there are others that have not been addressed. Garnering an understanding of each individual defect is not nearly as important as understanding the difference between cyanotic and acyanotic defects. Prehospital care for the CHD patient is primarily supportive, but, in certain cases, may require substantial intervention. EMS providers should not be concerned with diagnosing specific defects, but should be aware of the global effects that various defects have on normal perfusion. EMS providers' comprehension of the pathophysiology of CHDs and prompt actions will play a vital role in the outcome of the acutely ill CHD patient. PMID- 12889427 TI - Simulating the pediatric patient. PMID- 12889428 TI - Pediatric fever and febrile seizures. PMID- 12889429 TI - Developing a highly contagious disease readiness plan: the SARS experience. AB - The CDC and the World Health Organization have characterized a highly contagious syndrome that has resulted in numerous deaths worldwide. This infectious agent represents a new arena of highly contagious illness that could be a significant threat to the U.S. healthcare system. This disease has severely affected populations in the Far East and in Canada. While relatively few cases have been identified in the United States, there is a significant possibility that this agent will affect U.S. populations in the autumn--a time period when many viral diseases migrate from the Far East to North America. EMS agencies must use this window of time to define the characteristics of dangerous agents and develop a highly contagious disease readiness plan. PMID- 12889430 TI - Responding to SARS. PMID- 12889431 TI - EMS mythology, Part 5. EMS myth #5: Steroids are effective in the treatment of acute spinal cord injury. PMID- 12889432 TI - Is your station secure? AB - All department personnel must practice and assure safety and security of stations, vehicles, equipment and related items. Keep vehicle bay doors closed unless the bays are physically occupied by a department member. When responding or leaving the station, ensure, after exiting the bay, that the door is closed. If confronted with questions pertaining to department operations, including SOPs and SOGs, box alarms, response patterns, training times, member rosters/addresses, etc., do not provide the information. Document the incident and immediately report it. Should the inquiry appear extremely unusual in nature, do not hesitate to contact law enforcement. Emergency service personnel should be educated on a periodic basis and remain vigilant at all times. PMID- 12889433 TI - Stun guns. The medical implications. PMID- 12889434 TI - Spit-catcher: for people without mommies. PMID- 12889435 TI - EMS in Canada: Part 2. PMID- 12889436 TI - Telemedicine: laws still need a dose of efficiency. AB - Ever have a cold that just won't go away? You try lozenges, vitamin C, and a humidifier. Nothing seems to work. Telemedicine still has the same cold today in the form of laws and regulations that make the practice of telemedicine onerous. The best a health practitioner can do today is weather the cold that plagues telemedicine and make the best of it until legislators and regulators work on solutions to remove the barriers to effective telemedicine practice. This article provides a view of the landscape of telemedicine law today, outlines the barriers to the effective practice of telemedicine, and offers strategic concerns for health care providers to consider before entering into telemedicine arrangements. PMID- 12889437 TI - How to keep employees from taking you to court. AB - A lawsuit from a current or former employee, even an unfounded one, can have a devastating effect on a medical practice. In this article, the author suggests specific strategies for preventing employee lawsuits, specifically those that often result from mistakes in a practice's recruitment materials, employee handbook, job references, and dress code. In addition, this article offers guidance for preventing sexual harassment charges, overtime disputes, and hassles with problem employees. Finally, this article suggests four must-have business forms that provide important legal protection for any medical practice. PMID- 12889438 TI - Office space planning and design for medical practices: Part 2, Remodel or build new. AB - The decision to build anew or remodel/add-on to your existing facility can be a tough one. Your practice could possibly have multiple levels of ownership, equity, or lack of equity in the existing facility, emotional ties to the current location or building, etc. This article will deal with just how a practice makes that decision. PMID- 12889439 TI - Deploying the EMR: Part 2, Implementation. AB - This case study presents a collection of principles and guidelines regarding the process of selecting and deploying an electronic medical record (EMR) in a physician's practice. Practical advice and observations are detailed about what was learned and how the practice has benefited. In Part 1 of this article (Volume 18, pages 256-261), the authors identified the functions that are crucial in an ideal EMR. Part 2 provides their guidelines for implementing the system with particular emphasis on selection of the vendor. PMID- 12889440 TI - Computer briefs: laptop or desktop? PMID- 12889441 TI - Professional liability: ERISA's shield continues to erode. PMID- 12889442 TI - Beyond physician burnout: keys to effective emotional management. AB - Mismanagement of the complex emotions and work/life balancing challenges that come with a medical career is a risk factor that tends to go unaddressed in today's medical workplace. Visionary medical leaders recognize this dilemma and become ambassadors of effective emotional management (EEM) for physicians. EEM is a model for promoting stress resilience through management of personality-based stress reactions and the interpersonal consequences that come with coping habits. This article summarizes research that has shown how EEM affects physician, patient, and organizational functioning. It is proposed that training in EEM can serve as both the palliative and prophylactic intervention for physician distress, burnout, and the organizational problems it can cause. Taxonomy for assessing burnout is specified, and the special coping risks that come in the wake of malpractice litigation are discussed. Specific strategies from promoting EEM are detailed. PMID- 12889443 TI - Group practice contracting with managed care: Part 1. AB - Managed care contracts are complex and require understanding of market characteristics, legal considerations, and the needs of both the managed care organization and the practice. This article provides a framework for administrators and practices to consider when approaching negotiations for such contracts. PMID- 12889444 TI - Trends in the consumer price index. PMID- 12889445 TI - Streamline your practice with informational brochures. PMID- 12889446 TI - Surgery-related claims and the systems involved. AB - Patients--and, increasingly, plaintiffs' attorneys--seem to demand perfection from physicians, especially surgeons. But, the inevitability of human error demands that health care providers learn from mistakes and strive to reduce their recurrence. The recommendations provided within this article and those that follow can help surgical teams position themselves to address the individual and systems issues that are key to identifying harm and reducing adverse events. PMID- 12889447 TI - UK Radiation Oncology Conference(UKRO). University of Bath, United Kingdom. 7-9 April 2003. Abstracts. PMID- 12889448 TI - HIV therapeutics: points from the recent literature. PMID- 12889449 TI - Patients who want to stop their medications: treatment interruptions in HIV infection. PMID- 12889450 TI - NNRTI choice: has 2NN changed our practice? PMID- 12889451 TI - Triple diagnosis: dual diagnosis and HIV disease, Part 1. AB - Substance use disorders and psychiatric illness commonly co-occur in what is known as dual diagnosis. With the spread of HIV infection in persons with dual diagnoses, the triple diagnosis has emerged as a clinically challenging condition for primary care physicians, addiction medicine specialists, and psychiatrists. Existing data support the high prevalence of triple-diagnosis patients in psychiatric, substance abuse, and HIV treatment settings. This review highlights the features of substance abuse-psychiatric illness and its association with changes in antiretroviral therapy use, adherence, and HIV treatment outcomes. An integrated and interdisciplinary approach addressing substance abuse and mental health issues should be a primary element of comprehensive HIV care. Evaluating the safety and efficacy of psychopharmacologic and psychotherapeutic interventions and understanding the complex interactions among the components of the triple diagnosis are areas for future research. Service delivery and intervention models based on "1-stop shopping" should be developed and put into practice in order to optimize clinical outcomes. PMID- 12889452 TI - Fatal lactic acidosis and pancreatitis associated with ribavirin and didanosine therapy. AB - Pancreatitis and lactic acidosis are severe and life-threatening adverse events associated with nucleoside analogue antiretroviral therapy used to treat HIV infection. The drug from this class most commonly associated with these adverse events is stavudine, although zidovudine and didanosine have also been implicated. Ribavirin is a nucleoside analogue used in combination with interferon alfa to treat hepatitis C. Because of similar mechanisms of action, the combination of these 2 drugs could potentially increase such toxicity. A case of fatal lactic acidosis and pancreatitis is described in an HIV-infected patient coinfected wtih hepatitis C on a didanosine-containing antiretroviral regimen after treatment of hepatitis C was initiated with ribavirin and pegylated interferon alfa-2b. Extreme caution should be exercised when didanosine and ribavirin are used concomitantly because of the increased risk of mitochondrial toxicity and the syndrome of severe metabolic acidosis with elevated lactic acid levels. PMID- 12889453 TI - Editorial comment: drug-drug interactions, hepatitis C, and mitochondrial toxicity. PMID- 12889454 TI - Knee arthritis: flawed therapies. PMID- 12889456 TI - A sensory tune-up for your balancing act. PMID- 12889455 TI - Stacking up the benefits of three popular herbs. PMID- 12889457 TI - Eight simple steps to an agile mind. PMID- 12889458 TI - I recently developed proctitis after completing radiation treatment for prostate cancer. What should I do? PMID- 12889459 TI - Self-organized maps of sensory events. PMID- 12889460 TI - Percutaneous vertebroplasty: indications, technique, and results. AB - The authors describe a dedicated therapeutic vertebroplasty technique that uses newly designed instruments, acrylic cement, and dual guidance with ultrasonography and computed tomography for pain control in patients with bone failure, and report their experience. Between 1990 and 2002, they performed 868 percutaneous cementoplasty procedures in patients with severe osteoporosis, vertebral tumors, and symptomatic hemangiomas. In patients with osteoporosis, satisfactory results were obtained in 78% of cases; in patients with vertebral tumors, satisfactory results were obtained in 83% of cases; and in patients with hemangiomas, satisfactory results were obtained in 73% of cases. In the global series of 868 cementoplasties, an epidural leak was observed in 15 cases, which caused neuralgia in only three patients without spinal cord compression. In two patients, an asymptomatic pulmonary embolism was detected. The needle of the new vertebroplasty set is designed with side wings for easier rotation and removal. The screw syringe increases the precision of injection. The risk of leak is substantially reduced. The system is safe, reduces the cement manipulation time, and allows excellent control of the injection. The authors performed 130 vertebroplasties with this system without major complications. PMID- 12889461 TI - Navigating the aorta: MR virtual vascular endoscopy. AB - Virtual vascular endoscopy (VVE) uses two- and three-dimensional (3D) data sets from magnetic resonance (MR) or computed tomographic angiography to create endoluminal views of blood vessels. This technique is relatively new and has become practical only recently as cheap and powerful computers have become widely available. MR-generated VVE can produce striking images and may prove useful as an alternative or accessory means of presenting large quantities of data. This exhibit reviews technical aspects of MR VVE, describes common errors and artifacts, and provides several examples of MR VVE along with more traditional presentations of 3D gadolinium-enhanced MR angiographic data. PMID- 12889462 TI - Standardized evaluation of CT angiography with remote generation of 3D video sequences for the detection of intracranial aneurysms. AB - Computed tomography (CT) angiography is a well-known imaging technique commonly applied to both the detection and therapy planning of intracranial aneurysms. For this purpose, current studies predominantly focus on three-dimensional (3D) representations of CT angiographic volumes obtained with varying visualization approaches on different computers. Interactive manipulation performed by users individually is an important prerequisite for data analysis. However, this leads to inconsistent and barely reproducible 3D visualization results. Furthermore, the quality of any 3D representation depends on the applied visualization strategy (eg, maximum-intensity projection, shaded-surface display, direct volume rendering). To overcome these limitations, the authors present a novel method for standardized visualization of CT angiographic volumes, consisting of three steps: (a) transfer of the image data to a remote high-end graphics workstation, (b) automatic 3D visualization with high-resolution direct volume rendering, and (c) consecutive video generation performed according to a standardized protocol. The recorded video sequences are transferred for evaluation to a local desktop computer. In the experimental setup, high-quality videos based on 3D visualizations were produced in less than 60 minutes per patient. Although aneurysms above the skull base are usually visualized with excellent quality, the analysis of aneurysms at the skull base is still difficult. PMID- 12889463 TI - [50-year retrospectives of the system of health standards]. PMID- 12889464 TI - [Progress in research of pediatric nutrition in China]. PMID- 12889465 TI - Suicide rates in Russia on the increase. PMID- 12889466 TI - S Africa HIV drug report leaked. PMID- 12889467 TI - African countries to cooperate on epidemic control. Experts hope that sharing expertise and resources will help control disease outbreaks in the region. PMID- 12889468 TI - [Postoperative visual quality after corneal refractive surgery]. PMID- 12889469 TI - [Current status and development of the ophthalmology informatics]. PMID- 12889470 TI - [The influence of the anti-glaucoma operation on endodermic cell of cornea]. PMID- 12889471 TI - Alteration of vascular response to norepinephrine, calcitonin gene-related peptide, and acetylcholine in orchidectomized rats. AB - AIM: To determine the long-term effects of orchidectomy on vasoreactivity. METHODS: Orchidectomized rats with or without hormone replacement were raised 1, 2, and 4 months. Vascular responses to norepinephrine, calcitonin gene-related peptide (CGRP), and acetylcholine were tested on three different arteries (thoracic aorta, pulmonary artery, and caudal artery) by functional experiments. RESULTS: Responses to norepinephrine, CGRP, and acetylcholine were not changed by 1 or 2 months deprivation of androgen. In aorta, the dose-response curve of CGRP was shifted to the left by 1 month of testosterone replacement. Response to norepinephrine was increased 4 months after orchidectomy and reversed by testosterone replacement in thoracic aorta. Orchidectomy did not affect CGRP- and acetylcholine-induced relaxation in all the arteries 4 months after orchidectomy. CONCLUSION: Compared to estrogen, the protective effects of androgen on vasoresponsiveness may be through inhibiting vasoconstriction, but is relatively weak. PMID- 12889472 TI - Mitochondria and nitric oxide. PMID- 12889473 TI - Do home cholesterol tests work? PMID- 12889474 TI - The truth about irradiated meat. PMID- 12889475 TI - Waist management: gauging your risk. PMID- 12889476 TI - Snore silencers. PMID- 12889477 TI - Finance issue brief: mandated benefits: newborn hearing screening coverage requirements: year end report-2002. PMID- 12889479 TI - Finance issue brief: medical necessity: year end report-2002. AB - The information in this issue brief is based on a 50--state survey and a recent literature review. The Health Policy Tracking Service recognizes the complexity of this issue and discourages the use of this document as a sole resource on the issue. PMID- 12889478 TI - Endoglycan, a member of the CD34 family, functions as an L-selectin ligand through modification with tyrosine sulfation and sialyl Lewis x. AB - During lymphocyte homing to secondary lymphoid organs and instances of inflammatory trafficking, the rolling of leukocytes on vascular endothelium is mediated by transient interactions between L-selectin on leukocytes and several carbohydrate-modified ligands on the endothelium. Most L-selectin ligands such as CD34 and podocalyxin present sulfated carbohydrate structures (6-sulfated sialyl Lewis x or 6-sulfo-sLex) as a recognition determinant within their heavily glycosylated mucin domains. We recently identified endoglycan as a new member of the CD34 family. We report here that endoglycan, like the two other members of this family (CD34 and podocalyxin) can function as a L-selectin ligand. However, endoglycan employs a different binding mechanism, interacting with L-selectin through sulfation on two tyrosine residues and O-linked sLex structures that are presented within its highly acidic amino-terminal region. Our analysis establishes striking parallels with PSGL-1, a leukocyte ligand that interacts with all three selectins, mediating leukocyte-endothelial, leukocyte-leukocyte, and platelet-leukocyte interactions. Since the distribution of endoglycan includes hematopoietic precursors and leukocyte subpopulations, in addition to endothelial cells, our findings suggest several potential settings for endoglycan mediated adhesion events. PMID- 12889480 TI - Finance issue brief: point of service: year end report-2002. AB - A point-of-service (POS) option is a type of plan offered by managed care organizations (MCOs), including health maintenance organizations (HMOs), that allows people who are willing to pay higher out-of-pocket costs to see out-of plan providers. Mandating a point-of-service option essentially eliminates the use of closed-panel HMOs. This issue brief addresses only bills that mandate managed care plans to offer a point-of-service option. PMID- 12889481 TI - Finance issue brief: women's health: domestic violence anti-discrimination requirements: year end report-2002. PMID- 12889482 TI - Nucleoside-free regimen tested. PMID- 12889484 TI - Vaccine generates controversy. PMID- 12889483 TI - Teen girls confused about Pap smears. PMID- 12889485 TI - Gender and HIV website launched. PMID- 12889486 TI - HPV test combined with Pap. PMID- 12889487 TI - Microbicides get funding boost. PMID- 12889489 TI - Vaccine testing being combined. PMID- 12889488 TI - No staph outbreak in New York. PMID- 12889490 TI - Oral sex and HIV investigated. PMID- 12889491 TI - Canadian Thoracic Society COPD Guidelines: summary of highlights for family doctors. PMID- 12889492 TI - I've recently read that urban environments are associated with schizophrenia. Does city living really cause schizophrenia? PMID- 12889493 TI - [Reticulocyte indices in the differential diagnosis of iron-deficiency anemia and heterozygote beta-thalassemia]. PMID- 12889494 TI - Finance, pharmaceuticals issue brief: women's health: contraceptive coverage requirements: year end report-2002. PMID- 12889496 TI - Creating narratives in the clinical encounter. PMID- 12889495 TI - A challenging dialogue: a response to Arthur Frank. PMID- 12889497 TI - Finance issue brief: women's health: inpatient maternity stay coverage requirements: year end report-2002. PMID- 12889498 TI - Severe acute respiratory syndrome (SARS), a zoonotic infection with rapid spread: are we ready for the coming ones? PMID- 12889499 TI - Recent advance on viral hepatitis A. AB - Hepatitis A virus (HAV) infection is an old disease in the world, usually spread by the fecal-oral route. The uncomplicated patients usually recovery within 6 months. Its mortality is age-dependent. A high fatality rate with fulminate course and among chronic hepatitis B or C patients with HAV super-infection was observed. In recent two to three decades, the improvement of public health and hygiene has reduced the incidence of hepatitis A in children and adolescents worldwide; therefore, a massive outbreak of hepatitis A becomes possible among general population who neither have been infected nor have had any protective antibodies. Currently, the commercially available inactivated hepatitis A vaccine is proved to be safe and immunogenic in different risk groups, including children and adults, in several large clinical trials. Considering the potential for epidemic outbreak and the severity of the disease, immunization against the infection among risk population turns important for the prevention and control of HAV infection. PMID- 12889500 TI - Clinical characteristics of fatal patients with severe acute respiratory syndrome in a medical center in Taipei. AB - BACKGROUND: The clinical spectrum of severe acute respiratory syndrome (SARS) varies widely in the way that asymptomatic carriers are believed to exist in the community. Still there are severe forms of illness in which the patients deteriorate unexpectedly within hours. This study delineates clinical characteristics of such fatal cases of SARS for the purpose of identifying patients with poor outcome. METHOD: Patients with the diagnosis of probable or suspected SARS admitted to the Taipei Veterans General Hospital during the period from March 26, 2003 to May 25, 2003 were included. The medical records of fatal cases were retrospectively reviewed. RESULTS: During the study period, thirty-six probable cases and 17 suspected cases of SARS were identified. Eight probable but none of the suspected cases died from acute respiratory distress syndrome (ARDS) with multiple organ dysfunction after a median of 6-day hospital stay (range, 1 30 days). All but 2 patients acquired the infection nosocomially. Of the fatal cases, four were males and 4 females, with the median age of 65 years (range, 29 76 yrs). All except 3 had co-morbid conditions, such as hypertension, diabetes mellitus, coronary artery disease or chronic obstructive pulmonary disease. The lactate dehydrogenase (LDH) values were abnormal (> 200 U/L) in all patients. The chest radiograph of these fatal cases consistently showed multifocal infiltration over the unilateral or bilateral lobe of the lung. CONCLUSIONS: Patients of SARS who had advanced age, co-morbid conditions, highly elevated LDH and multifocal infiltration over chest radiograph should be closely monitored and actively treated. PMID- 12889501 TI - Management of hospital-acquired severe acute respiratory syndrome with different disease spectrum. AB - BACKGROUND: Outbreak of severe acute respiratory syndrome (SARS) in Taipei has been associated with Taiwanese back from Guangdong, China. We report 4 probable SARS cases with different severity and propose optimal treatment. METHODS: Four probable SARS cases were enrolled. Two cases were due to outbreak of SARS in our hospital and two cases were transferred from other hospitals. All patients received standard treatment: ribavirin 1000 mg orally daily for 10 days, Levofloxacin 500 mg orally daily for 7 days, and intravenous immunoglobulin (IVIG) 1 g/kg/day for 2 day after the onset of symptoms. If severe hypoxia (PaO2/FiO2 < 200) occurred, protective strategy of mechanical ventilation and methylprednisolone 2 mg/kg/day were given. The clinical pictures and treatment outcome were discussed. RESULTS: Fever, dyspnea, diarrhea, malaise, dizziness and dry cough were initially more common symptoms. Initially chest patterns included focal consolidation, interstitial infiltration or normal. Common laboratory findings were lymphopenia, and elevated serum levels of lactate dehydrogenase and C-reactive protein. No mortality was found. CONCLUSIONS: Highly alert and stringent infection control of SARS cases are required. Otherwise, SARS easily induces hospital-acquired first then community-acquired infection. Initial presentation of radiographic patterns includes normal, interstitial or airspace shadowing. Fever and lymphopenia are occasionally followed by rapidly progressive respiratory compromise. The standard treatment might be beneficial for decreasing the mortality rate. PMID- 12889502 TI - Dynamic cerebrovascular regulation in patients with autonomic dysfunction: a transcranial Doppler study. AB - BACKGROUND: Regulation of cerebral blood flow during orthostatic stress has been a major research interest. The purpose of this study was to scan healthy volunteers and patients with orthostatic dizziness and autonomic dysfunction for differences in cerebral hemodynamic patterns during orthostasis. METHODS: Thirteen patients with orthostatic dizziness and autonomic dysfunction and 9 healthy volunteers were recruited for monitoring of variations in intracranial hemodynamics with transcranial Doppler ultrasound during a 30-minute head-up tilt. Heart rate and blood pressure were monitored with surface electrocardiography and cuff sphygmomanometer, respectively. Cerebral blood flow velocity was continuously monitored using transcranial Doppler ultrasonography. RESULTS: The baseline mean cerebral blood flow velocity was significantly lower in the patient group (p < 0.05). After tilting up, the extent of immediate decreases in systolic blood pressure and in the mean blood flow velocity were significantly more in the patients (p < 0.05). However, the mean blood flow velocity remained significantly more retarded for up to 90 seconds after the initial drop in the patient group, while the blood pressure showed no significant difference between the patients and controls. CONCLUSIONS: Impaired dynamic cerebrovascular regulation can be documented as a delayed recovery of cerebral blood flow velocity upon orthostatic challenges, which may help verify the status of cerebrovascular regulation in patients with autonomic dysfunction. PMID- 12889503 TI - Outreach pharmacy service in old age homes: a Hong Kong experience. AB - BACKGROUND: To explore drug-related problems in old age homes in Hong Kong through outreach pharmacy service. METHODS: A standard form was used by outreach pharmacists to identify drug-related problems at old age homes. Homes were selected through random sampling, voluntary participation or adverse selection. Initial observation and assessment were performed in the first and second weeks. Appropriate advice and recommendations were given upon assessment and supplemented by a written report. Educational talks were provided to staff of the homes in addition to other drug information materials. At week 7 to 9, evaluations were carried out. RESULTS: Eighty-five homes were assessed and identified to have problems in the drug management system. These problems could generally be classified into physical storage (8.8%), quality of storage (19.2%), drug administration system (13.3%), documentation (16.4%), and drug knowledge of staff of homes (42.2%). Quality of drug storage was the most common problem found, followed by documentation and drug knowledge (73%, 50% and 44% of points assessed with problems, respectively). Apart from lack of drug knowledge and unawareness of potential risks by staff, minimal professional standards unmet may be fundamentally related to lack of professional input and inadequacy in legislation. Most homes demonstrated significant improvements upon simple interventions, from a majority of homes with more than 10 problems to a majority with less than 5 problems. CONCLUSIONS: Diverse problems in drug management are common in old age homes, which warrants attention and professional inputs. Simple interventions and education by pharmacists are shown to be effective in improving the quality of drug management and hence care to residents. While future financing of old age home service can be reviewed within the social context to provide incentives for improvement, review of regulatory policy with enforcement may be more fundamental and effective in upholding the service standard. PMID- 12889504 TI - Meconium peritonitis in utero: prenatal sonographic findings and clinical implications. AB - BACKGROUND: Prenatal ultrasound scans reveal variable features of fetal meconium peritonitis. For better understanding of the natural history and postnatal prognosis of meconium peritonitis, we clarified the significance of ultrasound findings in the outcomes of fetuses with meconium peritonitis. METHODS: Between 1991 and 2001, fetuses who underwent evaluation for meconium peritonitis were retrospectively analyzed. The diagnosis was made by intra-abdominal calcifications, meconium pseudocyst, or meconium ascites, which were postnatally confirmed by the operative findings and/or abdominal plain films. According to the final ultrasound findings before labor, patients were classified into 3 groups: type I, large meconium ascites; type II, a large pseudocyst; type III, intra-abdominal calcifications, small meconium ascites and/or a shrinking pseudocyst. RESULTS: Nineteen fetuses (6 boys and 13 girls) were collected. Mean gestational age at diagnosis was 29.1 +/- 5.3 weeks and mean follow-up before delivery was 7.5 +/- 6.2 weeks. Mean gestational age at birth was 36.8 +/- 6.2 weeks. Six fetuses (31.5%) were delivered before 37 weeks of gestation. Type I was found in 5 (26.3%), type II in 3 (15.8%), and type III in 11 (57.9%). Intra abdominal calcifications were identified in 15 cases (78.9%), polyhydramnios in 4 (21.1%), and oligohydramnios in 3 (15.8%). Cardiopulmonary resuscitation at birth was required in 2 cases (type I). Twelve cases (63.2 %) were spontaneous perforations. Of 7 other cases (36.8%) studied, 6 had intestinal atresia and 1 had associated defects resulting in bowel perforation. Postnatal follow-up revealed all but 2 (died of sepsis and prematurity, respectively) doing well. CONCLUSIONS: Our results indicate that detailed fetal ultrasound scans are crucial for providing a prenatal diagnosis and proper perinatal management of meconium peritonitis. From the viewpoints of our study, proper prenatal counseling and risk evaluation should be offered to the parents of the fetuses with meconium peritonitis. PMID- 12889505 TI - Voice rehabilitation in laryngectomees: comparison of daily-life performance of 4 types of alaryngeal speech. AB - BACKGROUND: We compared 4 kinds of alaryngeal speech available in Taiwan in the performance of daily-life tasks. The speech methods included pneumatic device (Taiwan tube), electrolaryngeal speech, esophageal speech, and tracheoesophageal prosthesis. METHODS: Questionnaires covering various communication situations in daily-life tasks as well as subjective feelings about using alaryngeal speech were distributed to members of The Taiwan Laryngectomees' Association and were collected mostly in April 2000 in their annual meeting. RESULTS: A total of 108 members responded to the questionnaire (totally 160 were questionnaires distributed). The mean age of respondents was 68.6 years, and the average time after surgery was 14.5 years. A pneumatic device was used by 58.8% of the responders, esophageal speech was used by 24.7%, an electrolarynx by 10.3%, and a tracheoesophageal prosthesis by 6.2%. No statistical difference was noted among patients using any of the 4 devices in terms of 13 communication tasks in daily life, except 4 questions such as the convenience of alaryngeal speech, embarrassment in face of strangers, the effort required to speak, and the discomfort in throat/neck/mouth when speaking. No statistical differences were noted among the 4 devices in terms of intelligibility. CONCLUSIONS: All 4 alaryngeal speech methods can serve most of the communication functions in daily life tasks with no differences in intelligibility. Limitations in loudness and tone were common problems. It is crucial for surgeons to understand the performance of various alaryngeal speeches in daily life during rehabilitation consultations with patients. PMID- 12889506 TI - A randomized prospective trial of bowel preparation for colonoscopy with Fortrans compared with bisacodyl. AB - BACKGROUND: An empty and adequate clean colon is a prerequisite of diagnostic or therapeutic colonoscopy. Our aim is to assess the efficacy and tolerance of bowel preparation for colonoscopy with Fortrans (polyethylene glycols) versus oral bisacodyl. METHODS: One hundred and four consecutive patients scheduled for colonoscopy were prospectively enrolled and allocated to Fortrans group or bisacodyl group by block randomization. RESULTS: Ninety-four patients were included for final analysis. Forty-five patients underwent preparation with Fortrans. The efficacy and tolerance of colon preparation were similar with both methods (p = 0.102, p = 0.202). The latency before the first bowel movement and total preparation time were shorter with Fortrans than with bisacodyl (p = 0.0001 and 0.001, respectively). Statistically significant differences were noted in changes of body weight, serum levels of sodium and calcium in patients taking bisacodyl (p = 0.049, 0.015, and 0.038, respectively). The taste of Fortrans was rated significantly better than that of bisacodyl (p = 0.002). Patients experienced more abdominal discomfort completing preparation with bisacodyl (p = 0.002). CONCLUSIONS: Fortrans and bisacodyl are equally effective and well tolerated in preparation for colonoscopy. However, Fortrans preparation takes less time and provides a better taste with less abdominal discomfort and less change in body weight as well as serum levels of sodium and calcium than bisacodyl preparation. PMID- 12889507 TI - Ganglioneuroma presenting as an asymptomatic huge posterior mediastinal and retroperitoneal tumor. AB - Ganglioneuroma is a rare, differentiated, benign and slow-growing tumor that commonly arises from sympathetic ganglion cells. Most of them are asymptomatic and found incidentally. We here report a quite rare case of silent huge ganglioneuroma growing in both posterior mediastinum and retroperitoneum occurring in a 3.5-year-old girl. The patient was relatively well before and incidentally found to have a huge chest mass by chest X-ray film at an episode of respiratory tract infection. Computed tomography showed a huge tumor extending from bilateral posterior mediastinum to the level of the adrenal gland in the retroperitoneum. Initially, neuroblastoma was highly suspected and 24-hour urine vanillyl mandelic acid was slightly elevated. Cytology by bone marrow aspiration revealed no tumor nests or clumps. Biopsy and pathology proved it as ganglioneuroma (GN). Due to too extensive involvement of the tumor and compression of the vital vessels, surgical removal became difficult. The family of the patient refused surgery due to there being no significant symptoms. Because of the potential for growth of unresectable GN and because the component of neuroblasts could not be completely excluded, the patient was still in dangerous status. The only thing we can do is to keep the family alert and continue regular follow-up. PMID- 12889508 TI - Mood disorder and secondary adrenal insufficiency after pituitary tumor operation and radiotherapy. AB - Psychological manifestations can be noted in patients with pituitary hypofunction or injuries after surgery and radiotherapy. We report on a 38-year-old female with acute adrenal insufficiency who presented with an affective disorder. The patient was brought to our emergency department because of general weakness and fever. Low plasma glucose concentration and low blood pressure were noted at arrival. She expressed mood change and persecutive thoughts. The brain MRI demonstrated no obvious brain damage except empty sella. The cortisol level was undetectable. Intravenous hydrocortisone was administered after the diagnosis of acute adrenal insufficiency. The patient improved physically after treatment, but social withdrawal and inappropriate affect persisted. We present the case and review the relevant literature to impress on physicians that the cognitive disorders in hypopituitarism can result from not only hormone insufficiency, but also the influences of pituitary tumor surgery and radiotherapy on neuropsychological functions. PMID- 12889509 TI - Primary lymphoma of the penis. AB - Primary penile lymphoma is a rare neoplasm. We report a case of this unusual clinical manifestation. A 70-year-old male presented with a painless penile mass for 6 months. Penile ultrasonography disclosed a huge tumor over the left corpus cavernosum. Pathologic examination of needle biopsy showed diffuse, small and T cell type malignant lymphoma. Whole-body computed tomography scan and bone marrow aspiration revealed no evidence of distant metastasis. We treated this patient with chemotherapy alone and achieved complete remission 2 months later. There was absence of tumor recurrence 20 months after treatment. We review the literature and discuss this uncommon lesion. PMID- 12889510 TI - Genetic verification of the role of CCK-AR in pancreatic proliferation and blood glucose and insulin regulation using a congenic rat carrying CCK-AR null allele. AB - A number of studies have indicated that cholecystokinin type A receptor (CCK-AR) plays a crucial role in postnatal pancreatic proliferation and blood glucose regulation through stimulating insulin secretion. The Otsuka Long-Evans Tokushima Fatty (OLETF) rat has been shown to possess poor pancreatic proliferation (PPP) capability after pancreatectomy (Px). Here we have constructed a congenic strain which introgressed an OLETF-derived 18.5 cM genomic fragment identified in our previous quantitative trait locus (QTL) analysis as a locus responsible for PPP into normoglycemic F344 genetic background The introgressed region includes CCK AR null mutation. After Px, the congenic rat showed weak pancreatic proliferation equivalent to that of the OLETF rat. Furthermore, post-surgery non-fasting blood glucose levels for the congenic rats are significantly higher in comparison with the F344 rats. At 28 days after Px, the congenic rats also showed lower blood insulin levels than the F344 rats. These results further provide the genetic evidence that 1) CCK-AR is essential for pancreatic regeneration; 2) impaired pancreatic proliferation mediates the development of hyperglycemia. PMID- 12889511 TI - Response in DNA ploidy of hepatocytes to tamoxifen and/or melatonin in vivo. AB - Tamoxifen is known to induce hepatocarcinogenesis in experimental animals and reversible chronic liver diseases in humans. Melatonin has been recently introduced as an oncostatic agent, especially for hormone-dependent tumors. This study was designed in order to investigate whether melatonin has an effect onthe tamoxifen-induced hepatotoxicity. Wistar albino rats were injected tamoxifen citrate intraperitoneally in three different doses (10 mg/kg and 20 mg/kg bw for 26 days; and 45 mg/kg bw for three days). Another group of animals were treated with melatonin once a week in addition to daily tamoxifen injections, whereas the third group received melatonin only. The control animals were injected an equal volume of diluent at corresponding intervals. At the end of the experimental period, the animals were sacrificed and the livers were prepared for the flow cytometric DNA analysis. DNA histograms were analyzed using the multicycle program. In experimental groups, all animals had aneuploid cell population. The difference in the diploid/ aneuploid ratio of each experimental group as compared to the control group according to Fischer's exact test was found to be highly significant (p < 0.002 MEL vs control; and p < 0.0001 for both TAM vs control and MEL+TAM vs control). Among the tamoxifen-injected animals, the proportion of multiploidy to aneuploid cell population was 17, similar to those treated solely with melatonin. Although the melatonin plus tamoxifen group had higher multiploidy percentage (38%), the difference was not statistically significant as compared to the tamoxifen (or melatonin) groups. No significant difference was noted between the animals which were treated with three different doses of tamoxifen. S-phase fraction percentage was significantly different in melatonin- and melatonin plus tamoxifen-injected animals with regard to controls, the degree of significancy being < 0.05 for both. According to our data, tamoxifen injections induced DNA aneuploidy, but did not stimulate proliferation in the liver as estimated by S-phase fraction. Melatonin, whether alone or in combination with tamoxifen, stimulated cell proliferation and produced aneuploidy. PMID- 12889512 TI - Epithelial maturation and extracellular matrix in reduced enamel epithelium. AB - OBJECTIVE: To evaluate epithelial-mesenchymal interactions in morphological homeostasis through histologically orientated observations on the reduced enamel in epithelium. STUDY DESIGN: Specimens were taken from impacted molar sites in 20 patients using the surgery. These were processed using standard histological, pathological and biochemical techniques. Slides were observed with a Nikon Eclipse E-600, Plan Apo piece, Microscopy, and photography was done using a Nikon Coolpix 990 digital camera. RESULTS: Radiolucent lesions displayed the supporting fibrous connective tissue wall with stratified squamous epithelium lining mostly. None of the specimens displayed prominent CAM 5.2, and anti-human K7 reacuvity, and PAS positive staining. CONCLUSION: While the results of this study suggest that the reduced enamel epithelium showed a lower tendency in cell cycle, it does indicate that a histologically oriented study of reduced enamel epithelium is needed for the understanding of epithelial-mesenchymal interactions in morphogenesis. PMID- 12889513 TI - Increased lipid peroxidation in the liver and kidney and their mitochondrial fractions following buthionine sulfoximine induced glutathione depletion in guinea pigs. AB - Malondialdehyde (MDA) and diene conjugates (DC) and vitamin C levels and the activities of glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) were determined in the liver and kidney and their mitochondrial fractions of guinea pigs 48 h after the injection of L-buthionine-(S,R)-sulfoximine (BSO), a glutathione (GSH) depleting agent. In BSO-induced GSH depletion, lipid peroxidation and SOD activities were found to be increased but GSH-Px activities did not change in the liver and kidney and their mitochondrial fractions. In addition, vitamin C levels remained unchanged in the liver and kidney homogenates. These results indicate that GSH depletion may influence oxidative stress in the liver and kidney and their mitochondrial fractions of guinea pigs. PMID- 12889514 TI - Plasma antioxidants and type 2 diabetes mellitus. AB - Reports indicate that some complications of diabetes mellitus are associated with increased activity of free radicals and accumulation of lipid peroxidation products. The organism's susceptibility to free radical stress and peroxidative damage is related to the balance between the free radical load and the adequacy of antioxidant defenses. In the present study, the relationship between plasma oxidants and antioxidants in diabetes mellitus was investigated. Thirty patients with type-2 diabetes mellitus were examined as well as twenty healthy controls (matched for age and sex against the diabetic patients). The plasma insulin and C peptide levels in the diabetic group were significantly lower (p < 0.001) than that of the control group. The mean plasma fructosamine, lipid peroxide, lipids and low-density lipoprotein cholesterol (LDL-C) levels were significantly high (p < 0.001) in the diabetic group compared to the control group. There were not any significant differences in the plasma high-density lipoprotein cholesterol (HDL C) levels between the patients and the control group (p < 0.001). The type-2 diabetes mellitus patients exhibited higher activities of plasma superoxide dismutase (SOD) than control values, whereas plasma glutathione peroxidase (GPx) activities were significantly lower. Our results suggest that there seems to be an imbalance between plasma oxidant and antioxidant systems in patients with type 2 diabetes mellitus. The estimation of plasma antioxidant levels and their replenishment by exogenous agents when necessary may be useful in the prevention of the diabetic complications. PMID- 12889515 TI - Tamoxifen inhibits arylamine N-acetyltransferase activity and DNA-2-aminofluorene adduct in human leukemia HL-60 cells. AB - Genetic regulation of acetyl coenzyme A-dependent N-acetyltransferase (NAT)and O acetyltransferase (OAT) activities may play an important role in the metabolic activation of arylamine chemicals and carcinogens. N-acetylation is thought to be the first step in arylamine metabolism. The enzyme responsible for N-acetylation is called NAT. In this study, synthetic non-steroidal antiestrogen tamoxifen was selected for determining the inhibition of arylamine NAT activity, gene expression (NAT mRNA) and DNA-2-aminofluorene adduct formation in human leukemia HL-60 cell line. The results demonstrated that tamoxifen did not affect the level of NAT mRNA in HL-60 cells. But the results also showed that NAT activity and 2 Aminofluorene-DNA adduct formation in HL-60 cells were inhibited and decreased by tamoxifen in a dose-dependent manner when the doses of tamoxifen up to 100 micro M. We also examined the standard steady-state kinetic analysis, and the data showed that tamoxifen may be an uncompetitive inhibitor to NAT activity in cytosols based on the decrease apparent values of Km and Vmax. This report is the first finding that tamoxifen inhibited human leukemia HL-60 cells NAT activity and DNA-2-aminofluorene on adduct formation. PMID- 12889516 TI - Stability, blood partition and protein binding of an antifibrotic agent, oltipraz. AB - The stability, blood partition and factors influencing the binding of oltipraz to 4% human serum albumin (HSA) were evaluated. Oltipraz was relatively stable in various pH (1-12) solutions for up to 48-h incubation, however, it was unstable in pH 13 solution and rat plasma and urine. Oltipraz reached an equilibrium fast (within 30 s mixing manually) between plasma and blood cells o f rabbit blood and the plasma-to-blood cells concentration ratios were independent of initial blood concentrations of oltipraz, 1 and 5 microg/ml; the ratios ranged from 0.908 to 1.004. The binding of oltipraz to 4% HSA was independent of oltipraz concentrations ranging from 1 to 100 microg/ml using an equilibrium dialysis technique: the mean value was 95.0%. However, the binding of oltipraz was dependent on HSA concentrations (the low concentrations, 3, 2, 1 and 0.5% of HSA caused an increase in the unbound fraction of oltipraz by 1.32, 1.98, 3.44 and 5.31 times, respectively, compared with the mean value from 4-6% HSA), incubation temperature (the unbound fractions at 37 degrees C and 22 degrees C increased 1.89 and 1.73 times, respectively, than that at 4 degrees C) and the buffer pHs (the unbound fractions were 6.36, 6.51, 5.60 and 4.63% for buffer pHs of 5.8, 6.4, 7.4 and 8.0, respectively). PMID- 12889517 TI - Hepatic gluconeogenesis in rats trained to eat a single meal daily. Role of eating periodicity and the amount of food ingested in the last meal. AB - Rats trained to eat a single daily meal (MF rats), from 8:00-10:00 a.m., increased food intake from the 1st to the 12th (125%) day of feeding training. In this work we compared the influence of the higher food ingestion in the last meal and feeding training on hepatic gluconeogenesis. Thus, rats at the 1st (MF(1st day-5g) group) and 13th day (MF(13th day-5g) group) of training, refed with a fixed amount of food (5g) were employed. In addition, a third group of MF rats, refed on day 12 with 75% (12g) of the food ingested by MF rats on the 13th day of the feeding training (MF(13th day-12g)) was included. The experiments were performed at 22 h after meal (8:00 a.m.). Our results demonstrated that feeding training had a crucial role in determining gluconeogenesis from pyruvate (5 mM). Additionally, gluconeogenesis from L-glutamine (5 mM) was influenced by periodicity of eating and the amount of food ingested in the last meal. In contrast, gluconeogenesis from L-alanine (5 mM) was not influenced by both factors. In conclusion, our findings suggested that the hepatic gluconeogenesis was influenced by food ingestion and/or feeding training depending of the substrate investigated. These effects on gluconeogenesis may have implications for use in diabetic regimens. PMID- 12889518 TI - Alterations in the erythrocyte antioxidant system of blood stored in blood bags. AB - In the present study, we measured the concentrations of reduced glutathione (GSH) and malonyldialdehyde (MDA) and the activities of glutathione peroxidase (GSH Px), glutathione S-transferase (GSH-S-T), superoxide dismutase (SOD), catalase (CAT) and glucose-6-phosphate dehydrogenase (G-6-PD) in erythrocytes obtained freshly from adult male donors which was preserved with CPDA-1 anticoagulant (citrate,phosphate, dextrose, adenine) on different days of storage. At the end of the study, storage-associated alterations in antioxidant activities were noted and discussed. GSH, GSH-Px, GSH-S-T, SOD, CAT and G-6-PD activities decreased, but erythrocyte MDA levels, as anindex of lipid peroxidation, increased during the storage period. According to our results, glutathione-dependent antioxidant systems in erythrocytes might be depleted during long storage in blood bags. PMID- 12889519 TI - Increased cardiac weight in interleukin-6 transgenic mice with viral infection accompanies impaired expression of natriuretic peptide genes. AB - Atrial natriuretic peptide (ANP) and B-type natriuretic peptide (BNP) regulate cardiac hypertrophy. We investigated ventricular alterations of ANP and BNP in interleukin-6 (IL-6) transgenic mice (TG) and wild type (WT) mice with or without viral infection. The ANP and BNP mRNA/GAPDH mRNA ratios in the ventricles of IL-6 TG mice were twice that of WT mice, but were not increased significantly by viral inoculation. In WT mice, both ANP and BNP responses were significantly increased in the ventricles of mice 10 days after encephalomyocarditis (EMC) viral inoculation. Cardiac weight in IL-6 TG mice was significantly greater than in WT 10 days after viral inoculation. Left ventricular wall thickness and the diameter of ventricular myocytes also were greater in IL-6 TG than WT after viral infection. Primary cultures of neonatal rat cardiac myocyte showed that IL-6 increased ANP and BNP mRNA expression in a dose-responsive fashion. In summary, overexpression of ANP and BNP occurs in the ventricles of IL-6 TG mice, along with increased cardiac weight after infection with EMC virus, and impaired responses in the expression of ANP and BNP. PMID- 12889520 TI - Interleukin 18 levels reflect the severity of acute pancreatitis. AB - The serum interleukin 18 (IL-18) levels were significantly increased in cases of acute pancreatitis, corresponding to the severity of the disease. The IL-18 levels were also significantly correlated with the serum tumor necrosis factor alpha (TNF-alpha) and bilirubin levels. The possible involvement of IL-18 in the development of liver dysfunction in acute pancreatitis is suggested. PMID- 12889521 TI - Presence of Vicia graminea or Vicia unijuga lectin-binding (Vgu) glycoproteins with and without Thomsen-Friedenreich (T) antigen in normal human seminal plasma. AB - Vgu glycoprotein (Vicia graminea lectin- or Vicia unijuga lectin-binding glycoprotein) has been reported as oncofetal antigen, which is found in many kind of tumor tissues, amniotic fluid and fetal membranes. In autoradiography with an 125I-labeled Vicia unijuga lectin (VUA) probe and an 125I-labeled Arachis hypogaea lectin (PNA, anti-T lectin) probe, seminal plasma samples of eight healthy men gave 2-7 Vgu glycoproteins without T antigen, 1-2 Vgu glycoproteins with Thomsen-Friedenreich antigen (T antigen) and 1-8 T-antigen glycoproteins, respectively. These results show that in addition to T-antigen glycoproteins, normal human seminal plasma contains Vgu glycoproteins and Vgu glycoproteins with T antigen as seminal plasma components as well as other tumor markers such as CA19-9, CA-125 and CEA. PMID- 12889522 TI - Retinol binding protein in plasma to evaluate the hepatotoxicity of rats treated with CCl4. AB - Retinol binding protein (RBP) in plasma of rats treated with carbon tetrachloride (CCl4) was monitored to clarify if RBP is available for the evaluation of the drug-induced hepatotoxicity. Blood was withdrawn by heart puncture at 0 hr and 12 hr after i.p. administration of CCl4 (0.2 ml/kg) to rats. Lactate dehydrogenase (LDH) and alanine aminotransferase (ALT) in plasma significantly increased at 12 hr after CCl4 administration, compared with the control, while RBP in plasma significantly decreased. On the other hand, albumin in plasma was unaffected at 12 hr after CCl4 administration. Thus RBP seems to monitor the different aspects in the drug-induced hepatotoxicity from LDH and ALT, and from the viewpoint of protein synthesis in the liver, to be more sensitively affected by the drug induced hepatotoxicity than albumin. PMID- 12889523 TI - Well maintained expression of CYP genes in sandwich-culturing hepatocytes: quantitative analysis using real-time PCR method. AB - Sandwich-culturing is an excellent hepatocyte culturing method in drug metabolism studies, however, its advantages for gene expression of cytochrome P450 (CYP) have not been evaluated so far. The present study was undertaken to determine the utilities of sandwich-culturing hepatocytes for evaluation of CYP genes expression. Hepatocytes from male rats were cultured for 5 days between two layers of type-I collagen gel (sandwich-culturing) or over type-I collagen gel (single gel culturing). To determine the expression of CYP genes rapidly and accurately, the time course study using real-time RT-PCR quantification was conducted in the present study, and CYP2B1, CYP2B2, CYP3A2, CYP3A9 and CYP3A23 genes were measured. Albumin secretion was also measured by ELISA to evaluate cell viability. Higher expression and excellent maintenance of all CYP genes were confirmed in sandwich-culturing hepatocytes than those in single gel culturing. Particularly, significant difference in the amounts of CYP genes expression was observed between both methods after 3 days culturing. Albumin secretion was also higher in sandwich-culturing after 3 days culturing, suggesting that the cell viability of hepatocytes was maintained. These results indicate that sandwich culturing method is much more advantageous than the ordinate method in maintaining the CYP gene expression and cell viability. PMID- 12889524 TI - Dosing time-dependent pharmacological effects of anti-metabolites for rat cardiac graft. AB - Antimetabolites such as methotrexete and 6-mercaptopurine have been shown to have circadian variations in their toxicities. However, chronopharmacological profiles of mizoribine (Miz) that is newly synthesized as an anti-metabolic agent for immunosuppression, have not been evaluated. In this study, we examined the dosing time-dependent alterations in the pharmacokinetics and pharmacodynamics of Miz. In addition, chronopharmacology of azathiopurine (Aza) was also evaluated to compare with that of Miz. Initially, Miz (10 and 20 mg/kg) or Aza (20 mg/kg) was orally administered at 8:00 hr or 20:00 hr for 3 weeks to rats. To reveal the dosing time-dependent difference of pharmacokinetics, Miz (20 mg/kg) was orally given at 8:00 hr or 20:00 hr and blood was obtained for 12 hours. Finally, Miz (20 mg/kg) or Aza (20 mg/kg) was administered at 8:00 hr or 20:00 hr to rats with heterotopic allogeneic heart grafts. The Miz group treated at 8:00 hr and Aza group treated at 20:00 hr showed severe myelosuppression compared with their each opposite dosing time. AUC of Miz in the morning trial was twice as high as that in the evening trial. The graft survival durations of the Miz- and Aza-treated groups were significantly longer than those of the respective control groups, but were not affected by dosing time of each agent. These results suggest that the toxicity, but not efficacy of Miz is varied with the dosing time. The chronotoxicological phenomenon of Miz might be, at least in part, explained by the dosing time-dependent difference in serum drug concentrations and apparent clearance. PMID- 12889525 TI - Thymoquinone-induced relaxation of guinea-pig isolated trachea. AB - The effect of thymoquinone (TQ), the main constituent of the volatile oil of black seed (Nigella sativa), on the guinea-pig isolated tracheal zig-zag preparation was investigated. TQ caused a concentration-dependent decrease in the tension of the tracheal smooth muscle precontracted by carbachol. The effects of TQ were significantly potentiated by pretreatment of the tracheal preparations with quinacrine, a phospholipase A2 inhibitor, nordihydroguiaretic acid, a lipoxygenase inhibitor and by pretreatment with methylene blue, an inhibitor of soluble guanylyl cyclase. On the other hand, the effects of TQ were not influenced by pretreatment of the tracheal preparations with indomethacin, a cyclooxygenase inhibitor, propranolol, a non-selective beta-adrenoceptor blocker or by the pretreatment with theophylline, an adenosine receptors antagonist TQ totally abolished the pressor effects of histamine and serotonin on the guinea pig isolated tracheal and ileum smooth muscles. The results of the present study suggest that TQ induced relaxation of precontracted tracheal preparation is probably mediated, at least in part, by inhibition of lipoxygenase products of arachidonic acid metabolism and possibly by non-selective blocking of the histamine and serotonin receptors. This relaxant effect of TQ, further support the traditional use of black seeds either alone or in combination with honey to treat bronchial asthma. PMID- 12889526 TI - No effect of cysteine on the pharmacokinetics of intravenous azosemide in rats with protein-calorie malnutrition by pretreatment with 3-methylcholanthrene. AB - The effects of cysteine on the pharmacokinetics of azosemide were investigated after intravenous administration of drug, 10 mg/kg, to male Sprague-Dawley rats pretreated with 3-methylcholanthrene fed on 23% protein diet (control rats) and 5% protein diet without (rats with protein-calorie malnutrition, PCM) or with (rats with PCMC) oral cysteine (250 mg/kg, twice daily starting from the fourth week) for 4 weeks. After intravenous administration to rats with PCM, the metabolites of azosemide excreted in urine and recovered from gastrointestinal tract decreased significantly than those in control rats, however, the plasma concentrations, total area under plasma concentration-time curve from time zero to time infinity (AUC) and time-averaged total body clearance (CL) were not significantly different between two groups of rats. It was reported that after intravenous administration of azosemide, 10 mg/kg, to rats with PCMC without pretreatment 3-methylcholanthrene, some pharmacokinetic parameters restored fully or more than the level of control rats; the time-averaged nonrenal clearance and apparent volume of distribution at steady state were comparable to those in control rats, but the terminal half-life and mean residence time were significantly shorter, AUC was significantly smaller, and time-averaged renal clearance and CL were significantly faster than those in control rats. However, the above mentioned effects of cysteine on the pharmacokinetic parameters of azosemide in rats with PCM were not observed with pretreatment with 3 methylcholanthrene. PMID- 12889527 TI - Blood partition and protein binding of a new neuroprotective agent for ischemia reperfusion damage, KR-31378. AB - The blood partition of KR-31378 between plasma and blood cells and the factors influencing the binding of the drug to 4% human serum albumin (HSA) using an equilibrium dialysis technique were evaluated. KR-31378 reached an equilibrium rapidly between plasma and blood cells of rabbit blood. The equilibrium plasma/blood cells concentration ratios were independent of initial rabbit blood concentrations of KR-31378, 1, 10 and 50 microg/ml; the values were in the range of 1.42-2.33. It took approximately 12-h incubation to reach an equilibrium between plasma and isotonic Soresen phosphate buffer of pH 7.4 containing 3% dextran ('the buffer'). The binding of KR-31378 to 4% HSA was dependent on HSA concentrations (the binding values were 25.3 and 32.0% for HSA concentrations of 2 and 5%, respectively), incubation temperature (the binding values were 48.8, 29.0 and 25.8% for 4, 22 and 37 degrees C, respectively), pHs of isotonic Sorensen phosphate buffer containing 3% dextran (the binding values were 17.7, 20.6, 22.8, 25.6 and 29.5% for buffer pHs of 5.8, 6.4, 7.0, 7.4 and 8.0, respectively) and alpha-1-acid glycoprotein (AAG) concentrations (the binding values were 25.6, 29.9, 34.4 and 50.3% for AAG concentrations of 0, 0.08, 0.16 and 0.32%, respectively). PMID- 12889529 TI - Therapeutic and toxic blood concentrations of more than 800 drugs and other xenobiotics. AB - In order to assess the significance of drug levels measured in clinical and forensic toxicology as well as for Therapeutic Drug Monitoring (TDM) it is essential that good collections of data are readily available. For more than 800 substances, therapeutic ('normal') and, if data was available, toxic, and fatal plasma concentrations as well as elimination half-lives were compiled in a table. The compilation includes data for hypnotics, benzodiazepines, neuroleptics, antidepressants, sedatives, analgesics, anti-inflammatory agents (e.g., NSAIDs), antihistamines, antiepileptics, betaadrenergic antagonists, antibiotics (penicillins, cephalosporins, aminoglycosides, gyrase inhibitors), diuretics, calcium-channel blockers, cardiac glycosides, antiarrhythmics, antiasthmatics, ACE-inhibitors, opiate agonists, and local anesthetics, among others. In addition, toxicologically relevant xenobiotics were listed. Data have been abstracted from published information, both compilations and primary sources and have been completed with data collected in our own forensic and clinical toxicology laboratories. Wherever possible, ranges for therapeutic plasma concentrations are expressed as trough concentration at steady state. The half life values given for each drug are chosen to represent the terminal log-linear phase at most. It is the purpose to rapidly assess the significance of drug levels for the therapeutic monitoring of patients, and to facilitate the diagnostic and clinical assessment in case of intoxications. PMID- 12889528 TI - High-performance liquid chromatographic analysis and pharmacokinetics of terazosin in healthy volunteers. AB - A high-performance liquid chromatographic (HPLC) analysis of terazosin in 1 ml of human plasma was developed using prazosin as an internal standard. The plasma sample was extracted with dichloromethane and ethylether and a 100-microl aliquot was injected onto the reversed-phase column. The mobile phase, 0.02 M sodium phosphate buffer:acetonitrile:tetrahydrofuran = 720:220:60 (v/v/v), was run at a flow rate of 0.8 ml/min and the column effluent was monitored using a florescence detector set at 370 and 250 nm for the emission and excitation wave numbers, respectively. The retention times for terazosin and prazosin were approximately 6.4 and 9.8 min, respectively, and the coefficients of variation of terazosin were generally low, below 6.4%. The present HPLC method was successful for the pharmacokinetic study of terazosin in healthy volunteers. Following oral administration of terazosin, 2 mg, to 20 healthy male volunteers, the area under the plasma concentration-time curve from time zero to time infinity was 421 +/- 71.8 ng h/ml and terminal half-life was 9.83 +/- 1.29 h. PMID- 12889530 TI - Dithranol reaction with nitroxide radicals in DMSO, a HPLC study. AB - Dithranol (1,8-dihydroxy-9-anthrone), an efficient drug for the topical treatment of psoriasis undergoes a complex chemical transformation after topical application. An absorption phase HPLC method has been developed and validated to follow the appearance of its oxidative products in a DMSO solution. In DMSO solution dithranol, chrysazin, and biantrone were monitored simultaneously by HPLC during autooxidation process, as well as in the presence of nitroxide radicals which increase the reaction rate. The kinetics of the very early stage of dithranol transformation is presented for the first time and discussed. Two unknown dithranol-derived intermediates were found and partially characterised. PMID- 12889531 TI - Determination of some micro- and macroelements in preparations made from peppermint and nettle leaves. AB - As, Ca, Cd, Cu, Fe, Mg, Mn, Pb and Zn were determined by ICP-MS and ET-AAS in preparations made from peppermint and nettle leaves after microwave high pressure digestion with nitric acid. In preparation from nettle leaves the Ca content was more than three times higher than in preparations made from peppermint leaves. Only very small differences (less than 10%) were observed for all the other elements. In all the samples investigated the content of inorganic elements were below the WHO limit (where existing). Variation coefficients ranged from 0.68% to 10.5% for ICP-MS measurements and from 1.48% to 10.0% for ET-ASS. PMID- 12889532 TI - Kinetic study of derivatives of phenylcarbamic acid enantiomers in rabbit blood serum using an on-line coupled column liquid chromatographic system. AB - An on-line coupled HPLC system is described for the kinetic study of the enantiomers of 1-methyl-2-piperidinoethylesters of 2-, 3- and 4 alkoxyphenylcarbamic acid in rabbit blood serum. The method involves three steps: (i) off-line pre-concentration and cleanup step, (ii) analytical separation of the racemate on a reversed-phase stationary phase, and (iii) separation of the enantiomers on a teicoplanin aglycone chiral stationary phase (Chirobiotic Tag). The limit of the determination with standard solutions was approximately 5.0 microg/ml. In vitro degradation studies of enantiomers have demonstrated differences in the concentration of the enantiomers after the treatment. The rate constants of R(-)- and S(+)-forms of enantiomers (the same as the position of alkoxychain) were, not significantly different. The number of carbon atoms had an influence on the degradation kinetics. PMID- 12889533 TI - TLC and HPLC characteristics of desacetylmatricarin, leucodin, achillin and their 8alpha-angeloxy-derivatives. AB - Five guaianolides, including two pairs of isomers, from a Hungarian taxon of the Achillea millefolium group were characterized analytically. Different chromatographic systems on TLC and HPLC were developed for the analysis of these compounds. TLC of leucodin, 8alpha-angeloxy-leucodin, achillin, 8alpha-angeloxy achillin and desacetylmatricarin was performed on silica gel using dichloromethaneacetone and cyclohexane-ethylacetate mixtures as mobile phases. HPLC on stationary phases as LiChrospher RP2, LiChrospher RP8, LiChrospher RP18e, Hypersil BDS C18 and Aquasil C18 required isocratic and gradient systems with different methanol-water mixtures as mobile phases. The presented RF values and retention times allow the identification of the respective 2-oxo-guaianolides which are marker substances for certain non-proazulene containing species. Their TLC and HPLC fingerprints are compared to those of proazulene containing species and are relevant for quality control. PMID- 12889534 TI - Rapid and sensitive HPLC method for the simultaneous determination of dorzolamide hydrochloride and timolol maleate in eye drops with diode-array and UV detection. AB - A rapid and sensitive HPLC method has been developed for the simultaneous determination of dorzolamide hydrochloride and timolol maleate. The drugs were monitored with a diode-array detector at two fixed wavelengths (lambda = 250.0 nm for dorzolamide hydrochloride and 300.0 nm for timolol maleate). Liquid chromatography was performed on a RP-YMC pack ODS A-132 C18 (5 microm, 15 cm x 6.0 mm) column and the mobile phase consisted of an acetonitrile: phosphate buffer (pH 2.5): methanol (5:85:10 v/v/v) mix and a flow rate of 1.2 ml x min( 1). The linearity of the method ranged between 4.0-45.0 microg x ml(-1) for dorzolamide hydrochloride and and 2.0-20.6 microg x ml(-1) for timolol maleate in binary mixture. The procedure was successfully applied to the determination of these compounds in pharmaceutical preparations and gave a high recovery, good accuracy and precision without any interference by the excipients. PMID- 12889535 TI - HPLC analysis of neo-clerodane diterpenoids from Teucrium chamaedrys. AB - A simple, rapid analytical method for the quantitative determination of nine neo clerodane diterpenoids was developed. The neo-clerodane diterpenoids present in the plant material and extracts were separated with an acetonitrile-water gradient at a flow rate of 1 mL per minute. The HPLC separation was performed on a Phenomenex Luna C18(2) (150 x 4.6 mm I.D., particle size 5 microm) reversed phase column with detection at 220 nm. The limit of detection was 0.24-0.90 microg/mL. The relative standard deviation (RSD) values for the determination of neo-clerodane diterpenoids in plant extracts were less than 3.20%. This is the first analytical method developed for qualitative and quantitative analysis of nine neo-clerodane diterpenoids by HPLC with PDA detection. PMID- 12889536 TI - Enhancer effects on in vitro corneal permeation of timolol and acyclovir. AB - The aim of this study was to evaluate the ability of two non-toxic skin penetration enhancers, N-methylpyrrolidone (NMP) and a positively charged phospholipid mixture (PS), to increase in vitro corneal permeation of timolol maleate (TM) and acyclovir (AC) in comparison with two corneal absorption promoters, polyethylene glycol octadecyl ether (Brij 78) and sodium taurocholate (TA). In vitro experiments were performed on corneas from albino rabbits which were mounted in a perfusion apparatus. The concentrations of the enhancers being tested were: Brij 78 1%, PS 1%, TA 1%, NMP 5%, NMP 10%. The safety of the enhancers being tested was assessed in vitro by determining their effects on corneal hydration and in vivo by means of a modified Draize test. Calculating the amount of drug permeated at different time points (90 and 180 min) we observed that TA, PS and NMP 5% significantly increased the cumulative amount of AC permeated after 90 min but only PS was effective after 180 min. TA, Brij 78 and PS were able to increase significantly the amount of TM permeated after 90 min but after 180 min only Brij 78 retained its effect. TA, Brij 78 and NMP 10% significantly increased the percent hydration levels (% HL) compared to the control while PS and NMP 5% did not affect % HL. The results of in vivo ocular tolerability studies showed that the enhancers which caused an in vitro increase of % HL produced in vivo conjunctival and/or corneal damages. The results of this study suggest that PS could be regarded as a potential corneal enhancer to increase the intraocular bioavailability of AC and TM. PMID- 12889537 TI - Biopharmaceutical characterization of some synthetic purine drugs. AB - Hydrophilic-lipophilic properties (water solubility, n-octanol/water partition coefficient, transport across membranes) of some mercaptopurines (6-MP, 6-TG, AZA and a new AZA derivative--metazathioprine (MAZA) were determined. MAZA is the most lipophilic compound due to low aqueous solubility and high n-octanol/water partition coefficient. The fluxes from the donor medium into the membrane and from the membrane into the acceptor medium are highest for MAZA as well. The partition coefficients of the other purines decrease in the order: AZA > 6-TG > 6 MP. PMID- 12889538 TI - Interaction of quinidine, disopyramide and metoprolol with melanin in vitro in relation to drug-induced ocular toxicity. AB - The aim of this work was to evaluate binding capacity of quinidine, disopyramide and metoprolol to melanin in vitro. The antiarrhythmics studied cause adverse reactions to the eye. Synthetic DOPA-melanin was used in the studies and a UV spectrophotometric method was employed to determine the drugs. The studies of the kinetics of the formation of quinidine-melanin, disopyramide-melanin and metoprolol-melanin complexes indicate that for all the complexes investigated the maximum time to reach reaction equilibrium is 24 h. Binding parameters, i.e., the numbers of independent binding sites and the association constants were determined on the basis of the Scatchard plots. An analysis of the binding curves obtained supports our conclusion that both strong (n1) and weak (n2) binding sites are involved in the formation of the complexes investigated. The total numbers of binding sites in synthetic DOPA-melanin complexes with quinidine, disopyramide and metoprolol were 0.525, 0.493 and 0.387 micromol/mg, respectively. The quinidine-melanin complex is characterized by greater stability (K1 = 3.00 x 10(5) M(-1), K2 = 1.75 x 10(3) M(-1)) in comparison with biopolymer complexes with disopyramide (K1 = 1.12 x 10(4) M(-1), K2 = 6.04 x 10(2) M(-1)) and metoprolol (K1 = 1.42 x 10(4) M(-1), K2 = 7.89 x 10(2) M(-1)). The ability of these drugs to form complexes with melanin in vitro may be one of the reasons for their ocular toxicity in vivo, as a result of their accumulation in melanin in the eye. PMID- 12889540 TI - Triterpene constituents from the leaves of Melicope indica. AB - Phytochemical investigation of a petroleum ether extract of Melicope indica afforded two unusual pentacyclic triterpenes viz. neohop-13(18)-en-3alpha-ol (1) and fern-8(9)-en-3beta-ol (2) and the ubiquitous steroids, stigmasterol and sitosterol. The structures of 1 and 2 were independently elucidated on the basis of 2D NMR data and confirmed by comparison with those of related compounds. While compound 1 is a new natural product, this is the first report of occurrence of fern-8(9)-en-3beta-ol (2) from the genus Melicope. PMID- 12889539 TI - Chemoprotective effects of ethanolic extract of neem leaf against MNNG-induced oxidative stress. AB - We evaluated the modifying effects of ethanolic extract of neem leaves (Azadirachta indica A. Juss) on oxidative stress induced by the potent gastric carcinogen N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) in male Wistar rats. The extent of lipid peroxidation and the status of the antioxidants superoxide dismutase (SOD), catalase (CAT), reduced glutathione (GSH), glutathione peroxidase (GPx) and glutathione S-transferase (GST) were used as intermediate endpoints of chemoprevention. Three different concentrations of ethanolic neem leaf extract (100, 200 and 400 mg kg(-1) body weight) were administered by intragastric intubation (i.g) for five consecutive days followed by MNNG (i.g) 1.5 h after the final administration. Enhanced lipid peroxidation was accompanied by compromised antioxidant defences in the stomach, liver and erythrocytes of MNNG-treated rats. Pretreatment with ethanolic neem leaf extract at a dose of 200 mg/kg body weight (bw) significantly lowered the concentration of lipid peroxides and increased antioxidant levels. Our results demonstrate that neem leaf exerts its chemoprotective effects on MNNG- induced oxidative stress by decreasing lipid peroxidation and enhancing the antioxidant status. PMID- 12889541 TI - Formula optimization based on neural networksin sustained release tablets. PMID- 12889542 TI - Antimutagenic activity of aqueous extracts and essential oil isolated from Myrtus communis. PMID- 12889543 TI - Healing vs. fixing: how Ellie learned to dance. PMID- 12889544 TI - Examining the ethics of praying with patients. AB - Nurses should reflect carefully about the meaning and purpose of prayer in the clinical setting before engaging in prayer with patients. This article discusses the ethics of prayer with patients in regard to respectful care of the patient and integrity for the nurse. Five guidelines are offered to assist nurses in their ethical decisions about prayer with patients. PMID- 12889545 TI - Prayer's clinical issues and implications. AB - Because prayer frequently sustains coping and brings comfort, it is an important resource for nurses to support or offer. How shall nurses incorporate prayer in nursing practice? This article explores practical aspects of including prayer in bedside nursing care, including suggestions for assessment, supporting patients when beliefs about prayer challenge, discussing prayer with clients for whom prayer may be harmful, overcoming nursing barriers to prayer, and more. PMID- 12889546 TI - Large clinical study shows value of therapeutic touch program. AB - Representing the largest published sample size of therapeutic touch (TT) outcomes to date, data from this continuous quality improvement (CQI) clinical study suggests that TT, when provided in the clinical setting, promotes comfort, calmness, and well-being among hospitalized patients. In addition, patients are highly satisfied with TT. The newly developed Patient Satisfaction Survey and TT Performance Improvement Tool provide an effective means by which to evaluate a TT program. PMID- 12889547 TI - Sleep quality in children with juvenile rheumatoid arthritis. AB - Children with juvenile rheumatoid arthritis (JRA) report poor sleep quality, daytime sleepiness, fatigue, anxiety, and altered mood. Sleep disturbances in school-aged children are an issue of serious concern. Children are at an age when sleep is of primary importance to physical and intellectual growth, and sleep disturbances that begin in childhood may persist into adulthood. In this article we will review what is currently known about sleep in children with JRA, the influence of medications on sleep quality, the potential impact of poor sleep quality on daily life issues, and complementary/alternative modalities that may be effective in reducing sleep disturbances. PMID- 12889548 TI - Integrating mindfulness-based stress reduction. AB - Mindfulness-based stress reduction (MBSR) programs may mitigate the effects of stress and disease. This integrative review identified 21 clinical studies on MBSR interventions. Although preliminary findings suggest health enhancement from MBSR, controlled, randomized studies, the operationalization of constructs, and qualitative research are needed. PMID- 12889549 TI - Reiki therapy: the benefits to a nurse/Reiki practitioner. AB - This study evaluates how nurses who gave Reiki therapy perceived the benefit of this therapy on their clients and on themselves concurrently as providers of the therapy. As an adjunct, the study's purpose was to enhance the understanding and credibility of nurse/Reiki practitioners. PMID- 12889550 TI - JCAHO safeguards spiritual care. PMID- 12889551 TI - Constipation in idiopathic Parkinson's disease. PMID- 12889552 TI - Cell kinetic study on histogenesis of Barrett's esophagus using rat reflux model. AB - BACKGROUND: To elucidate the histogenesis of Barrett's esophagus and esophageal adenocarcinoma, we designed a duodeno-gastric reflux model in which normal stomach function and normal nutritional status are retained. METHODS: Male Wistar rats were used in the experiment. The esophago-gastric junction was side-to-side anastomosed to a loop of jejunum about 3 cm distal to Treitz's ligament. The animals were not exposed to any known carcinogens during the experiment. Sequential morphological changes were studied for up to 50 weeks after surgery. Serial sections were made and stained with hematoxylin and eosin (H&E). In addition, immunohistochemical staining for bromodeoxyuridine (BrdU) was performed along with histochemical staining for mucins using paradoxical concanavalin A (ConA), galactose oxidase Schiff (GOS), and high-iron diamine-alcian blue (HID AB). RESULTS: Severe esophagitis with squamous cell hyperplasia was noted in all animals after surgery. At week 20 after surgery, glandular metaplastic cells positive for ConA first appeared within the basal cell layer of esophageal squamous cell epithelium, and then GOS-positive cells and HID-AB goblet cells appeared. This is a characteristic of the specialized columnar epithelium of Barrett's esophagus. We detected esophageal adenocarcinomas in 1 out of 8 subjects at week 40 and in 3 out of 8 subjects at week 50 after surgery. CONCLUSIONS: Reflux of duodenal contents causes specialized columnar epithelium of Barrett's esophagus and esophageal adenocarcinoma. As part of the sequence of events leading to the development of Barrett's esophagus, pyloric-foveolar metaplasia was observed followed by the appearance of intestinal goblet cells. The pyloric-foveolar metaplasia appears to be associated with chronic mucosal damage and regeneration. This multiplastic cell lineage is referred to as 'gut regenerative cell lineage' (GRCL). PMID- 12889553 TI - Low frequency of upper gastrointestinal complications in a cohort of high-risk patients taking low-dose aspirin or NSAIDS and omeprazole. AB - BACKGROUND: There is very little information available on the incidence of complications and on the best prevention therapy in high-risk patients taking non steroidal anti-inflammatory drugs (NSAIDs) and/or aspirin. Randomized-controlled trials in such patients are rare for ethical reasons. We studied the incidence of gastrointestinal complications in high-risk patients taking long-term low-dose aspirin or non-aspirin-NSAIDs combined with omeprazole in a real-life clinical setting. METHODS: This was a multicentre, prospective and observational study including 247 consecutive high-risk patients who had a clinical indication for long-term treatment with either low-dose aspirin or non-aspirin NSAIDs and omeprazole therapy. The occurrence of gastrointestinal complications was measured. RESULTS: In addition to a recent history of peptic ulcer bleeding, all patients had at least 1 other risk factor and 112 (45.3%) had 3 or more risk factors; 78.9% were taking low-dose aspirin and the remainder non-aspirin NSAIDs. Mean follow-up was 14.6 +/- 10.38 months. Three patients taking low-dose aspirin developed upper gastrointestinal bleeding (1.2%; 95% CI 0.3-3.5; 1.0 event/100 patients/year). This was similar to the rate observed in studies involving non high-risk patients taking low-dose aspirin and higher than that observed in patients not taking low-dose aspirin. Two additional patients developed a lower gastrointestinal bleeding event (0.81% (0.04%-3.12%); 0.67 events/100 patients/year), which was within the range expected in NSAID users. CONCLUSIONS: The use of omeprazole in the high-risk patient taking low-dose aspirin or NSAIDs seems to be a safe therapeutic approach in this population and is associated with a low frequency of upper gastrointestinal complications. PMID- 12889554 TI - Histopathological parameters of Helicobacter pylori-associated gastritis in children and adolescents: comparison with findings in adults. AB - BACKGROUND: The pathohistological features of Helicobacter pylori-associated gastritis in children and adolescents are less well understood than they are in adults. The aim of the study was to compare histological parameters of H. pylori infected children with those of adults. METHODS: The retrospective study compared histological features of 111 children (mean age 10.8 +/- 3.8). Three paediatric age groups were analysed and the findings were compared with those of 111 adults (mean age 64.2 +/- 12.1). Degree of chronicity and activity of inflammation, mucus depletion and regeneration of foveolar epithelium by regenerating epithelium and H. pylori colonization were scored in antral biopsies. RESULTS: The histological parameters in children, i.e. degree of chronicity, activity of gastritis and the summed gastritis score, were not significantly different compared to those in adults. Replacement of foveolar epithelium by regenerating epithelium was significantly larger in adults compared to that of paediatric patients. The rate of low-grade mucus depletion and of the strongest degree of H. pylori colonization was higher in children than in adults. Children with antral nodularity had significantly higher histological score values. CONCLUSION: The histological differences between paediatric patients and adults are focused on signs of chronic inflammation and regeneration. Our results imply that antral nodularity is an important sign of highest-grade gastritis, especially in young children. PMID- 12889555 TI - Comparative classification and grading of Helicobacter pylori gastritis in patients with gastric cancer and patients with functional dyspepsia. AB - BACKGROUND: It has previously been shown that corpus-dominant grade and activity of Helicobacter pylori gastritis in combination with intestinal metaplasia in the antrum or corpus are risk markers for the development of stomach cancer. If one point is scored for each of these three parameters, a gastric cancer risk index is obtained that permits prediction of the risk of gastric cancer developing on the soil of H. pylori gastritis. The aim of the present study was to evaluate the accuracy of the gastric cancer risk index based on a large number of patients compared with dyspeptic controls. METHODS: In 415 biopsied patients with gastric carcinoma, biopsy specimens taken from the antrum and corpus were investigated retrospectively. From this group of patients, 244 patients positive for H. pylori were compared with 244 sex- and age-matched H. pylori-infected patients with functional dyspepsia. RESULTS: H. pylori gastritis was detected in 395 carcinoma patients (95.2%). The 244 sex- and age-matched patients significantly more frequently had corpus-dominant H. pylori gastritis (compared with NUD controls). The incidence of intestinal metaplasia was also significantly increased. For a gastric cancer risk index score of 3 points (i.e. corpus pronounced grade and activity of gastritis, and intestinal metaplasia in antrum or corpus), a sensitivity of 93% and a specificity of 85% for the presence of gastric carcinoma can be calculated. CONCLUSION: Using the proposed risk index, the topographic grading of H. pylori gastritis in the antrum and corpus enables the diagnosis of a 'risk gastritis' to be made. PMID- 12889556 TI - Fish oil feeding is associated with an increased accumulation of dietary lipids in enterocytes: results from an in vivo study in rats. AB - BACKGROUND: Chronic fish oil consumption is associated with reduced postprandial lipaemia, but the mechanism behind this effect is not fully understood. We studied whether lipid absorption might be altered in rats fed fish oil. METHODS: Male Wistar rats were fed fish oil enriched chow (n = 6) or control oil enriched chow (n = 6). After 4 weeks, 61 mg 3H-triolein was instilled into duodenal tied off loops. Intestinal segments were removed after 15, 30, 45, 60 and 90 min. Enterocytes were then isolated by calcium chelation and quantified by DNA determination. Non-absorbed 3H-lipid and 3H-lipid contents of enterocytes were determined by liquid scintillation counting. Two other groups of rats (2 x 6) fed the experimental diets were given an oral fat load and fasting and postprandial blood samples were taken. RESULTS: The accumulation of 3H-lipids in enterocytes was higher in rats fed fish oil than in controls (area under the 3H-lipid time curve: 1041.3 versus 670.3 nmol oleic acid x min/microg DNA, P < 0.05). Separation of lipids showed that an accumulation of triglycerides and free fatty acids occurred in rats fed fish oil. The amount of non-absorbed 3H-lipid tended to be higher in the fish oil fed rats (P > 0.1). It was confirmed that the fish oil enriched chow caused lower postprandial lipaemia (34% reduction in serum triglyceride concentrations, P < 0.05). CONCLUSION: Attenuated postprandial lipaemia following fish oil feeding is explained, at least partly, by a transient lipid accumulation in enterocytes which may result in a delayed triglyceride efflux from the enterocytes into the circulation. PMID- 12889557 TI - Luminal salmonella endotoxin affects epithelial and mast cell function in the proximal colon of pigs. AB - BACKGROUND: Salmonellosis and systemic endotoxaemia affect intestinal function. However, little is known about the functional importance of luminal Salmonella (S.) endotoxin during intestinal infection. METHODS: Pigs were either given or not given lipopolysaccharide (LPS, 30 mg day(-1)) of S. Typhimurium DT-104 orally for 14 days. Blood samples were taken weekly. After slaughter (day 14), epithelia of the proximal colon were investigated in Ussing chambers. Bacterial translocations to lung, liver, spleen and several lymph nodes were determined by culture. RESULTS: Endotoxin feeding increased plasma C-reactive protein (CRP) and histamine levels without evoking clinical signs. Postmortem, proximal colonic epithelia of LPS-treated animals showed both a decreased histamine release after mast cell stimulation with A23187 and a smaller increase in short-circuit current after A23187 application. Addition of the nitric oxide donor, sodium nitroprusside (SNP), also elicited lower increases in short-circuit current in the proximal colon of endotoxin-treated pigs. Endotoxin pre-feeding decreased colonic ion conductance, although mannitol and histamine fluxes were high in some epithelia of this group. Luminal Salmonella endotoxin increased bacterial translocation to proximal jejunal lymph nodes. LPS applied to colonic epithelia in vitro had no electrophysiological effects. CONCLUSIONS: Luminal endotoxin elicits an acute phase response and affects intestinal electrolyte transport and mast cell function. Furthermore, LPS induces epithelial spots of increased mannitol permeability that could be identical to spots of enhanced bacterial translocation. PMID- 12889558 TI - Effectiveness of the sorbitol H2 breath test in detecting histological damage among relatives of coeliacs. AB - BACKGROUND: Small intestinal lesions have a wide severity in coeliac disease (CD), and early diagnosis is important in preventing neoplastic and non neoplastic disorders related to CD. The aim of this study was to compare the effectiveness of the sorbitol H2 breath test (H2-BT) and serological tests (antigliadin (AGA), antiendomysium (EMA) and anti-tissue transglutaminase (anti tTG)) as screening tests in the detection and estimation of CD prevalence in 1st degree relatives. METHODS: Screening was performed in 111 1st-degree relatives of 37 coeliac families. Sorbitol H2-BT, AGA, EMA and anti-tTG antibodies were used to select the candidates for small-bowel biopsy. Relatives with abnormal serological tests and/or with sorbitol H2-BT positivity underwent a small-bowel biopsy. Small-bowel biopsy was also performed in relatives negative in all tests but with clinical complaints or suspected of having CD, and intestinal lesions were expressed according to the Marsh classification. RESULTS: CD was diagnosed in 49/111 screened relatives (44.14%): 5 showed Marsh IIIc, 8 Marsh IIIb, 16 Marsh IIIa, 13 Marsh II and 7 Marsh I lesions. Nineteen relatives showed the classical form of the disease, while the subclinical and silent forms were recorded in 20 and 10, respectively. AGA, EMA and anti-tTG showed strong positivity only in severe intestinal damage (Marsh IIIb-c lesions) (but overall positivity was 36.73%, 38.78% and 44.89% for AGA, EMA and anti-tTG, respectively), while sorbitol H2-BT showed strong positivity also in patients with slight histological damage (Marsh I-IIIa) (overall positivity was 83.67%). CONCLUSIONS: A significant proportion of coeliacs may be missed if relatives are screened by serology only, while the efficacy of sorbitol H2-BT in screening relatives is confirmed. This study confirms that neither a breath test nor serology can replace intestinal biopsy, which remains the gold standard for the diagnosis of CD. PMID- 12889559 TI - Applicability of the Ussing chamber technique to permeability determinations in functionally distinct regions of the gastrointestinal tract in the rat. AB - BACKGROUND: Ussing chambers are commonly utilized for in vitro investigations into gastrointestinal permeability. However, their sensitivity and applicability to the small intestine have not been well characterized. METHODS: In order to investigate the effects of experimentally induced damage and the relative contribution of the mucosa and muscularis externa layers to transmural permeability in the small intestine, stomach and colon, normal rat intestinal tissues were mounted in Ussing chambers with or without removal of the muscularis externa or mucosal layers. Gastric tissues were damaged in vivo by exposure to indomethacin (100 mg kg(-1)), while ileal tissues were damaged in vitro by 0.4 M NaCl. Tissue damage was assessed histologically, while permeability parameters included conductance (G), potential difference (PD) and mucosal to serosal flux of horseradish peroxidase (HRP). RESULTS: Damage localized to the tissue edges (edge damage) accounted for 25%-50% of the exposed epithelial length in the ileum, while less than 20% of stomach and colon epithelium was affected by edge damage. In the damaged stomach, a 20% reduction in epithelialization was accompanied by increases in G (P < 0.001) and HRP (P < 0.01) flux. Removal of the muscularis externa did not affect mucosal viability in the undamaged ileum or colon although HRP flux in the colon, but not ileum, was increased (P < 0.01). Removal of the ileal mucosa produced increases in G and HRP flux, while PD was maintained. CONCLUSION: We conclude that the Ussing chamber technique is suitable for application to studies of gastric and colonic permeability in rats. However, owing to the prevalence and extent of edge damage in the small intestine, we would caution against the use of this technique for permeability studies in this region of the gastrointestinal tract in the rat. PMID- 12889560 TI - Antibodies to oat prolamines (avenins) in children with coeliac disease. AB - BACKGROUND: The use of oats in a gluten-free diet for children with coeliac disease is presently under investigation. In this study we measured the content of antibodies to oat prolamines (avenin) in sera from coeliac children and reference children. METHODS: Crude avenin was prepared by extraction with ethanol and salt-solution and used as antigen in a three-step ELISA. Sera from 81 children, including 34 children with verified coeliac disease, were analysed for both IgA and IgG antibodies to avenin and gliadin. Sera were also incubated with gliadin before exposure to avenin, and vice versa, to assess a possible cross reaction between the species. Keyhole limpet hemocyanin (KLH) was used as a negative control. RESULTS: Children with coeliac disease on a normal diet had significantly higher levels of antibodies to avenin, both IgG and IgA, than reference children (P < 0.001) and the levels correlated positively with gliadin antibodies, especially of IgA-type (r = 0.798). Both anti-avenin and anti-gliadin antibodies were only absorbed by the corresponding protein. CONCLUSIONS: Children with coeliac disease have antibodies to oat proteins at significantly higher levels than reference children. The absorption test did not indicate a cross reactivity between the prolamines of wheat and oats. The method will be employed for repeated sampling of anti-avenin antibodies during a prospective interventional study with a gluten-free diet supplemented with oats. PMID- 12889561 TI - Prevalence of coeliac disease: unexplained age-related variation in the same population. AB - BACKGROUND: The aims of this work were (a) to evaluate the prevalence of coeliac disease (CD) in a large sample of the Brazilian general population and (b) to compare CD prevalence between children and adults. METHODS: The study group comprised 4405 subjects (2629 F and 1776 M). Age distributions were 2034 (1-14 years), 848 (15-29), 584 (30-44), 667 (45-59) and 272 above 60. The immunoglobulin A antiendomysial antibody (IgA-EMA) test was used as the serological screening tool. All sera were submitted to turbidimetric measurement of IgA levels and those with IgA deficiency to the IgG antigliadin (IgG-AGA) test. The small intestinal biopsy was recommended for subjects showing either (a) IgA-EMA positivity or (b) selective IgA deficiency (SigAD) and IgG-AGA positivity. RESULTS: There were 16 EMA positive out of 4405 sera tested. SigAD was found in five cases (one adult and four children). Two of these children tested positive for IgG-AGA and underwent jejunal biopsy that, in both cases, disclosed a normal mucosa. Overall, 17 out of 18 eligible subjects performed the small intestinal biopsy. The prevalence of biopsy-proven CD in this study group was 3.41 per 1000 individuals. If all 18 EMA-positive patients were included, the overall prevalence would become 3.63 per 1000. The prevalence in adults and children was 2.11 per 1000 and 5.44 per 1000, respectively. CONCLUSION: This work supports previous findings showing that CD is not a rare disorder in Brazil and that there is an unexplained difference in the prevalence of CD between adults and children. PMID- 12889562 TI - Better dietary compliance in patients with coeliac disease diagnosed in early childhood. AB - BACKGROUND: In coeliac disease (CD) there is a permanent gluten intolerance requiring life-long adherence to a strict gluten-free diet (GFD). An inadequate diet increases the risk for long-term complications. Coeliac patients often have great difficulty in maintaining a strictly GFD. We aimed to study whether young adults with CD diagnosed before the age of 4 years have a better dietary compliance than patients diagnosed later in life. METHOD: Twenty-nine adults with CD diagnosed in childhood were studied. They had had CD for 17-24 (mean 20) years. Their compliance to GFD was assessed using a questionnaire and serological markers (IgA and IgG anti-endomysium antibodies and IgA anti-tissue transglutaminase antibodies). RESULTS: At least 80% of the coeliac patients who had been diagnosed before the age of 4 years complied with the GFD compared to 36% of the CD patients older than 4 years at diagnosis (P < 0.05). CONCLUSION: This is the first study to show that patients with CD diagnosed before 4 years of age keep to a GFD significantly better than patients diagnosed after 4 years. It is thus important to diagnose childhood CD as early as possible in order to minimize the risk for reduced well-being and other potentially serious complications in coeliac individuals on an inadequate diet. PMID- 12889563 TI - Reducing psychological distress in patients with inflammatory bowel disease by cognitive-behavioural treatment: exploratory study of effectiveness. AB - BACKGROUND: This prospective study aimed to determine whether cognitive behavioural group treatment accompanying medical standard care is effective in reducing psychological distress in patients with inflammatory bowel disease. METHODS: Twenty-eight outpatients with Crohn disease or ulcerative colitis completed the treatment programme. Psychological treatment consisting of 12 weekly sessions was conducted in a group setting. Medical and psychometric assessments were taken at the beginning of the 3-month pretreatment waiting period, at pretreatment, at post-treatment and at the 3, 6 and 9-month follow ups. RESULTS: During baseline, no change was observed in psychological distress. Disease-related worries and concerns decreased significantly from pretreatment to the follow-ups. The disease groups differed in the decline of concerns between pre- and post-treatment, with a significant reduction of concerns in patients with ulcerative colitis but not Crohn disease. This difference did not occur at the follow-ups, indicating long-term improvement for both disease groups. Depressive coping decreased significantly in women and remained stable at the follow-ups, whereas depressive coping did not change in men. The same gender difference was found for depressive symptoms. CONCLUSIONS: The exploratory findings suggest that psychological group treatment for outpatients is a feasible and effective approach for the short- and long-term reduction of psychological distress for patients with inflammatory bowel disease. However, the revealed gender differences on coping and depression might indicate the necessity to consider gender-specific aspects of inflammatory bowel disease when designing and evaluating psychological interventions. PMID- 12889564 TI - A group-based patient education programme for high-anxiety patients with Crohn disease or ulcerative colitis. AB - BACKGROUND: The inflammatory bowel diseases (IBD) ulcerative colitis (UC) and Crohn disease (CD) affect a person's health-related quality of life (HRQOL). IBD patients report high levels of anxiety, which correlates with the degree of perceived dissatisfaction with the information on disease-related themes provided in routine health care. The aim of this study was to evaluate changes in anxiety after participation in a group-based educational intervention for IBD patients screened for high anxiety. METHODS: The programme consisted of 8 sessions, and 49 patients participated. Anxiety was assessed using the Hospital Anxiety and Depression (HAD) Scale at baseline and 6 months after intervention. HRQOL was assessed with the Inflammatory Bowel Disease Questionnaire (IBDQ) and the SF-36 health survey. Participant satisfaction with education was measured using a study specific questionnaire. RESULTS: No significant change on the HAD anxiety score was found at the 6-month follow-up for those who participated in the education programme despite the fact that the participants reported they had gained better knowledge of disease-related items. Furthermore, there were no significant changes over time regarding bowel symptoms, systemic symptoms, emotional functioning and social functioning of the IBDQ or generic HRQOL (SF-36). CONCLUSIONS: IBD patients with a high anxiety level reported improved satisfaction with information about disease-related items, but did not indicate any benefits in terms of reduced anxiety or improved HRQOL after participating in the education programme, not at least in the short-term perspective. In this selected group of patients, psychosocial problems other than disease-related concerns were found that warrant other approaches. PMID- 12889565 TI - Comparison of percutaneous acetic acid injection and percutaneous ethanol injection for hepatocellular carcinoma in cirrhotic patients: a prospective study. AB - BACKGROUND: Ultrasound-guided percutaneous ethanol injection (PEI) and percutaneous acetic acid injection (PAI) are effective in the treatment of hepatocellular carcinoma (HCC). We conducted a prospective study to compare the therapeutic efficacy of both these methods. METHODS: Sixty-three patients were treated by PAI using 50% acetic acid and 62 by PEI using pure ethanol. There were no significant baseline differences in age, sex, Child-Pugh class, tumour size and number, or other clinico-biochemical parameters between the two groups. RESULTS: During a follow-up period of 24 +/- 9 (range 6-38) months, 19 (30%) of the PAI group and 21 (34%) of the PEI group died (P = 0.704). The 1- and 3-year survival rates were 84% and 51% for the PAI group and 81% and 46% for the PEI group (P = 0.651). The corresponding tumour recurrence rates were 51% and 74% for the PAI group, and 54% and 64% for the PEI group (P = 0.787). The treatment sessions were 3.9 +/- 1.6 and 6.2 +/- 2.3 for the PAI and PEI groups, respectively, in each treatment cycle (P = 0.008). A multivariate analysis using the Cox regression model revealed that ascites (relative risk (RR) 3.1, 95% confidence interval (CI) 1.5-6.3, P = 0.002), large (>3 cm) or multinodular HCCs (RR 2.4, 95% CI 1.1-5.4, P = 0.04), and development of tumour recurrence (RR 7.0, 95% CI 3.1-16.0, P < 0.001) were independent, poor prognostic factors in both groups. CONCLUSIONS: PAI and PEI are equally effective in the treatment of HCC. PAI has the advantage of fewer treatment sessions in each treatment course. Careful pretreatment patient selection may improve survival. PMID- 12889566 TI - Serum leptin levels in patients with liver cirrhosis and chronic viral hepatitis. AB - BACKGROUND: The aim of the present study was to investigate serum leptin levels in relation to anthropometric features in patients with liver cirrhosis (LC) and chronic viral hepatitis (CVH), and to determine the effect of the severity and aetiology of the LC on serum leptin levels. METHODS: Forty-nine patients with LC, 32 patients with CVH and 69 control subjects were age, body mass index (BMI) and sex-matched and included in the study. Plasma glucose, serum leptin and insulin levels were determined. Insulin resistance was assessed using homoeostasis model assessment (HOMA). Body composition was estimated by skinfold thickness. RESULTS: Female patients with Child-A LC had higher levels of leptin, and female and male patients with Child-A LC had higher absolute leptin (leptin/BFM) levels compared to patients with Child-C LC and control subjects. Serum leptin levels of the patients with alcohol LC were higher than the control subjects, but the absolute leptin levels were comparable. When alcoholic and post-viral hepatitis cirrhotic patients were compared with each other on an aetiologic basis, there was no significant difference between them in leptin and absolute leptin levels. There were significant correlations between leptin and BMI, body fat percentage (BFP), BFM (body fat mass) in all three groups in both sexes. CONCLUSIONS: These data suggest that the physiologic correlations among serum leptin level, sex, BMI and BFM were well preserved in patients with chronic liver disease. Patients with alcohol LC had higher leptin levels. In early stages of liver disease, leptin levels and absolute leptin levels are higher than in normal subjects. However, in advanced stages of the disease the significant decline in leptin levels and similar levels of leptin expressed in relation to BFM compared to control subjects predominantly represent the expression of fat mass. PMID- 12889567 TI - Gossypol activates pancreatic polyamine catabolism in normal rats and induces acute pancreatitis in transgenic rats over-expressing spermidine/spermine N1 acetyltransferase. AB - BACKGROUND: The male antifertility agent gossypol has been reported to induce spermidine/spermine N1-acetyltransferase (SSAT) in canine prostate cells. As SSAT is the rate-controlling enzyme in the catabolism of the polyamines and is involved in the development of acute pancreatitis in a recent transgenic rat model, we exposed normal and transgenic rats over-expressing SSAT to gossypol to evaluate its effect on pancreatic polyamine metabolism and organ integrity. METHODS: Pancreatic SSAT activity, polyamine pools, pancreatic histology and plasma 2-amylase activity were determined after different doses of gossypol. RESULTS: Gossypol increased pancreatic putrescine and decreased spermidine and spermine pools in normal rats accompanied by tissue oedema and significantly elevated plasma amylase activity. In transgenic rats, the drug strikingly induced SSAT, profoundly depleted the higher polyamines and caused distinct pancreatitis. The combination of gossypol at doses harmless to transgenic pancreas with an inhibitor of polyamine oxidase caused massive synergistic induction of SSAT, profound depletion of the polyamine pools and acute pancreatitis. CONCLUSIONS: The results indicate that gossypol induces pancreatitis through an activation of polyamine catabolism. PMID- 12889568 TI - Alcoholic, but not biliary, pancreatitis varies seasonally in occurrence. AB - BACKGROUND: Alcohol is the most common cause of acute pancreatitis in Finland (70%). The amount of alcohol consumed has been shown to be associated with the prevalence of pancreatitis in the country, and also to be an important determinant of the severity of the first episode of acute alcoholic pancreatitis. We have a clinical suspicion that the use of alcohol and the incidence of pancreatitis are increased during holiday periods in summer, although no seasonal variations have been reported in a German population. METHODS: Between 1972 and 1992 a total of 1556 episodes with acute alcoholic pancreatitis were treated at Tampere University Hospital; 552 were first episodes and 1004 were recurrent. For comparison, we investigated 297 episodes of acute biliary pancreatitis treated in that same time period. We investigated the monthly prevalence of acute alcoholic (first and recurrent) pancreatitis and compared this with the monthly absolute alcohol consumption. RESULTS: Taking all alcoholic pancreatitis episodes into account, significant differences can be seen between prevalence and month of onset of the disease (P < 0.0001), whereas among biliary pancreatitis episodes there were no differences (P = 0.3). Prevalence of acute alcoholic pancreatitis was significantly higher than the expected prevalence in July and August, and the amount of alcohol consumption (100% alcohol, litres) was highest during these same months. Also during March, October and December the prevalence was higher than expected. CONCLUSION: Months with holiday seasons, Christmas, Easter, summer and autumn, are associated with the highest alcohol consumption and the highest prevalence of acute alcoholic pancreatitis. PMID- 12889569 TI - Distribution of glucagon-like peptide-2 in normal colonic tissue. PMID- 12889570 TI - Adverse reactions to pantoprazole. PMID- 12889571 TI - Somatostatin therapy ameliorates chronic and refractory gastrointestinal bleeding caused by diffuse angiodysplasia in a patient on anticoagulation therapy. AB - We present a patient receiving chronic anticoagulant treatment with recurrent and intractable gastrointestinal bleeding due to diffuse angiodysplasia. Following failure of previous medical and surgical treatment, and in light of the patient's need for chronic anticoagulation due to mechanical heart valve, she was treated with somatostatin analogue, octreotide s.c. 100 microg on alternate days for 28 months. Treatment significantly decreased the occurrence of bleeding episodes, the need for hospitalization and blood transfusion requirements despite continued anticoagulant therapy. Octreotide treatment should be considered in patients with refractory gastrointestinal bleeding due to angiodysplasia in particular in those who need anticoagulant treatment. PMID- 12889572 TI - Upper GI cancer in patients with uncomplicated dyspepsia. PMID- 12889573 TI - Reinvesting in social justice: a capital idea for public health nursing. PMID- 12889574 TI - Postcolonial nursing scholarship: from epistemology to method. AB - Postcolonial theory, with its interpretations of race, racialization, and culture, offers nursing scholarship a set of powerful analytic tools unlike those offered by other nursing and social theories. Building on the foundation established by those who first pointed to the importance of incorporating cultural aspects into nursing care, nursing scholarship is in a position to move forward. Critical perspectives such as postcolonialism equip us to meet the epistemological imperative of giving voice to subjugated knowledges and the social mandates of uncovering existing inequities and addressing the social aspects of health and illness. This article makes a case for the integration of postcolonial perspectives into theorizing and sketches out a research methodology based on the postcolonial tradition. PMID- 12889575 TI - The nursing shortage as a community transformational opportunity. AB - The nursing shortage is continuing toward its projected peak in 2010. As a profession we seem unable to devise effective strategies to enhance growth and overcome this looming crisis. By focusing on predictable barriers using methods of reductionism, nurses may have forgotten the biggest barrier: the challenge of creating a holistic professional group presence and inclusive nursing stance. Without first addressing and solving internal paradoxical identity struggles, nursing will continue to try to overcome the multiple internal and external barriers that resulted in the current shortage. This article discusses the group dynamics that have fostered the cyclic loop of nursing shortage situations and explores the reflection and inquiry needed to formulate a cohesive national nursing group posture. PMID- 12889576 TI - Upstream reflections on environmental health: an abbreviated history and framework for action. AB - Upstream thinking considers the social, economic, and environmental origins of health problems that manifest at the population level. The upstream thinking perspective is applied to an examination of environmentally associated health problems and the opportunities that citizens have (or do not have) to access information and resources to make health-promoting choices in response to environmental health risks. A proposed framework for nurses to reduce environmental health risks includes distributive and strategic actions. Distributive actions include tracking, embedding, and translating; strategic actions include discovering through etiologic research, discovering through community-based research, advocating, and reframing. Together these actions can help formalize nursing's role in responding to citizens' concerns about environmental health problems. PMID- 12889577 TI - Caregiving Effectiveness Model evolution to a midrange theory of home care: a process for critique and replication. AB - A clinically relevant model, grounded in nursing theory, has evolved to become a midrange theory. This article describes the processes used to derive, validate, revise, and test the Caregiving Effectiveness Model. Testing of this midrange theory used prospective longitudinal research with family members caring for patientsrequiring lifelong, complex, technology-based home care. It presents the conceptual critiques and statistical procedures and discusses derivation of model generated nursing interventions and implications for use of these validation processes in developing nursing knowledge. The article summarizes limitations of the model and presents recommendations for future research. PMID- 12889578 TI - The concept of protection: a dimensional analysis and critique of a theory of protection. AB - Dimensional analysis clarifies the concept of protection, which has a commonly understood definition but is used inconsistently in research literature. Concepts such as protective factors and protective behaviors are often used interchangeably without adequately representing the phenomenon of protection itself. This article critiques a situation-specific theory of protection and presents dimensions of a model with an ecological view of protection. It uses dimensional analysis methods to derive the social construction of protection from its use in a broad range of literature; vigilant management and vigilant communication are salient dimensions of protection. The article compares conceptual literature with research literature to identify inconsistencies in use. PMID- 12889579 TI - Suffering: toward a contextual praxis. AB - Janice M. Morse's article in Advances in Nursing Science (24:1) revised and summarized the major findings of a research program exploring the behavioral experiential nature of suffering. Using a feminist critical theory stance, this article addresses Morse's conceptualization of a praxis of suffering. First, it identifies the strengths and contributions of Morse's body of research to nursing science. Next, it undertakes a critique situated in feminist critical theory in which the limitations of Morse's work are explored using exemplars from the Western literary tradition. Finally, the article proposes a new conceptualization of an alternative contextual praxis of suffering in which nurses' responses to suffering are situated in an emancipatory paradigm of authentic presence. PMID- 12889580 TI - Acute gastrointestinal bleeding. AB - Acute gastrointestinal bleeding is a common disorder with a wide spectrum of presentations that may encompass multiple clinical scenarios. Initial hemodynamic assessment and resuscitation are critical. Once accomplished, the source of bleeding should be localized, active bleeding should be stopped, the underlying abnormality should be treated, and recurrent bleeding should be prevented. The means to accomplish these goals depends on the specific clinical situation. For most forms of upper gastrointestinal bleeding, early endoscopy is the cornerstone of diagnosis and management. It can predict and improve clinical outcomes. A variety of endoscopic and pharmacologic modalities are effective at achieving and maintaining hemostasis. The optimum means of evaluation and treatment of acute lower gastrointestinal bleeding is less clear and is now evolving. Endoscopy (usually expectant, less often early) is widely used and effective for diagnosis but has unproven therapeutic benefits. Angiography is effective (diagnostically and/or therapeutically) in certain situations. Surgery offers the opportunity for definitive therapy at the cost of higher morbidity. At this time, the approach to evaluation and management should be based on the specific clinical situation and available local expertise. PMID- 12889581 TI - Acute intestinal ischemia and infarction. AB - Acute intestinal ischemia is a gastrointestinal emergency resulting from a sudden decrement in intestinal blood flow. It may occur as a consequence of mesenteric vascular occlusion and/or hypoperfusion and may involve the small intestine or colon. Bowel infarction, sepsis, and death may result, making prompt diagnosis and management imperative. Acute mesenteric ischemia generally stems from interruption of blood flow within the superior mesenteric artery or vein, and leads to small intestinal hypoperfusion and infarction. It carries with it a mortality rate of approximately 70%, but improved survival may be achieved as a result of early diagnostic consideration, undelayed angiography, and surgical intervention, when appropriate. Acute colonic ischemia occurs typically as a result of a transient mismatch between intestinal blood flow and the metabolic demands of the colon. Although infarction may occur, colonic ischemia is often a reversible condition with mortality rates considerably lower than those witnessed in acute mesenteric ischemia. This article reviews the pathophysiology, clinical features, diagnostic, and therapeutic options applicable to patients with acute intestinal ischemia. PMID- 12889582 TI - Biliary emergencies: pancreatitis, cholangitis, and more. AB - The most common cause of acute pancreatitis is gallstones, although many other etiological factors have been identified. The management of the initial episode depends on the severity of the attack and the etiology. In most patients, acute pancreatitis has a benign, self-limited course. However, in the minority who develop infected pancreatic necrosis the mortality can reach 25%. The early assessment of severity and aggressive management of these patients is critical. The roles of endoscopic retrograde cholangiopancreatography, surgical intervention, enteral feeding and use of antibiotics in acute pancreatitis are discussed in this article. Finally, the origin of recurrent acute pancreatitis is discussed, with particular reference to conditions such as pancreas divisum and sphincter of Oddi dysfunction whose role in the development of acute pancreatitis is controversial, and to hereditary or familial pancreatitis. PMID- 12889583 TI - Fulminant hepatic failure. AB - The term "fulminant hepatic failure" (FHF) encompasses a pattern of clinical symptoms and pathophysiological responses associated with rapid arrest of normal hepatic function. The syndrome is defined by the presence of hepatic encephalopathy in association with coagulopathy and jaundice. In many cases, the clinical picture is complicated by cerebral edema, renal impairment, sepsis, and multiorgan failure. In this review, we examine the specific causes of FHF, including acetaminophen-related hepatotoxicity, drug- and viral-related FHF, and other less common causes of FHF such as pregnancy and vascular related disease. The approach to FHF should be multidisciplinary and requires a thorough understanding of the biochemical, metabolic, and physiological changes associated with hepatic necrosis. Also examined are management issues pertinent to these complex situations and the role of liver transplantation and liver assist devices are considered. PMID- 12889584 TI - Emergencies after liver transplantation. AB - Liver transplantation has become the procedure of choice for a wide variety of patients with end-stage liver disease. Perioperative morbidity and mortality have decreased dramatically over the past two decades, and superior graft and patient survival rates are now routine. Despite these advances, however, there remain several potentially lethal possibilities that may complicate the immediate postoperative period. Failure of the graft to regain any useful metabolic activity is known as primary nonfunction, and almost uniformly requires retransplantation for any hope of survival. Lesser degrees of immediate dysfunction require experienced clinical judgment as to the probability of sustaining long-term patient viability. Another potentially catastrophic development is thrombosis of the grafted hepatic artery. This is sometimes successfully managed by surgical reconstruction. It may develop immediately, or present insidiously much later. Thrombosis of the portal vein, while not usually fatal, can significantly complicate the immediate course, carrying with it a significant risk of sepsis. Close monitoring of patients in the period following liver transplantation is crucial, as prompt diagnosis and early intervention directly affects the patient's chances of survival. PMID- 12889585 TI - What we have learned about the predictors of preterm birth. AB - The Preterm Prediction Study conducted by the Maternal Fetal Medicine Network between 1993 and 1996 studied a large number of risk factors for preterm birth in more than 3,000 women at 10 centers. The goals of the study were to better understand the strength of one risk factor versus another and to explore interactions among the predictors looking for combinations of factors that were more predictive of preterm birth than any single factor used alone. The most potent factors that were associated with spontaneous preterm birth at < 32 weeks were a positive cervical-vaginal fetal fibronectin test (odds ratio, 32.7) and < l0th percentile cervical length (odds ratio, 5.8), and in serum, > 90th percentiles of alpha-fetoprotein (odds ratio, 8.3) and alkaline phosphatase (odds ratio, 6.8), and > 75th percentile of granulocyte colony-stimulating factor (odds ratio, 5.5). Results for spontaneous preterm birth at < 35 weeks were generally similar but not as strong. The overlap among the strongest biologic markers for predicting spontaneous preterm birth was small. This suggests that the use of tests such as maternal alpha-fetoprotein, alkaline phosphatase, and granulocyte colony-stimulating factor as a group or adding their results to fetal fibronectin and cervical length test results may enhance our ability to predict spontaneous preterm birth and that the development of a multiple-marker test for spontaneous preterm birth is feasible. PMID- 12889586 TI - What we have learned about cervical ultrasound. AB - The National Institute of Child Health and Human Development Maternal-Fetal Medicine Units Network has completed 2 prospective, blinded observational studies of cervical ultrasound for the prediction of preterm birth and the identification of high-risk women who might benefit from interventions aimed at preventing spontaneous preterm birth. These reports and other clinical data support the concept that cervical competence is not a dichotomous variable, but more likely functions along a continuum that is reflected by the relationship between cervical length and reproductive outcomes. Because of its safety, availability, patient acceptance and reproducibility, the sonographic evaluation of the uterine cervix has become an important investigational tool that has advanced our understanding of the preterm birth syndrome and should allow a more focused approach to the most persistent and challenging problem in modern obstetrics. PMID- 12889587 TI - What have we learned about uterine contractions and preterm birth? The HUAM Prediction Study. AB - Measurement of uterine contraction frequency has been employed as a screening test to identify women with increased risk of preterm birth, and as an aid in the early diagnosis of preterm labor. The National Institute of Child Health and Human Development Maternal-Fetal Medicine Units (NICHD MFMU) Network performed a prospective, blinded observational study of uterine contraction frequency to detect and predict preterm labor and birth, respectively. The goal of the study was to assess the sensitivity, specificity, and positive and negative predictive value of various measures of uterine contraction frequency. Data collected from 306 women revealed that contraction frequency was significantly greater in women who would ultimately deliver before rather than after 35 weeks' gestation. However, both sensitivity and positive predictive value of any measure of contraction frequency to predict preterm birth were poor. Contraction frequency did not increase significantly within 1 or 2 weeks of an episode of preterm labor. These results serve to explain the absence of an association between contraction-based surveillance and preterm birth in randomized trials conducted in women at risk of preterm birth. PMID- 12889588 TI - What have we learned about vaginal infections and preterm birth? AB - Asymptomatic maternal genital tract infection during pregnancy, particularly bacterial vaginosis, has been consistently associated with preterm birth. In response to this evidence, the Maternal-Fetal Medicine Units Network (MFMU) designed and conducted 2 large randomized, placebo-controlled clinical trials of metronidazole treatment of asymptomatic pregnant women with bacterial vaginosis or trichomoniasis in a general obstetrical population. These studies showed that treatment of women with bacterial vaginosis failed to prevent preterm birth, regardless of their history of prior preterm birth. Metronidazole treatment of women with trichomoniasis significantly increased the risk of preterm birth compared to placebo. These results formed the basis of the US Preventive Services Task Force recommendation that screening for bacterial vaginosis not be undertaken in low-risk pregnant women, and show that MFMU network studies can have a direct and immediate impact on obstetrical practice. PMID- 12889589 TI - What we have learned regarding antibiotic therapy for the reduction of infant morbidity after preterm premature rupture of the membranes. AB - Preterm premature rupture of the membranes (pPROM) is responsible for approximately one third of the over 450,000 preterm births occurring in the United States annually. In this manuscript, we summarize the outcomes and analyses related to the National Institute of Child Health and Human Development Maternal Fetal Medicine Units Network (NICHD-MFMU) network multicenter trial of antibiotics to reduce infant morbidity after pPROM. Based on evident reduction in gestational age dependent and infectious infant morbidity, we provide the rationale for aggressive intravenous and oral, broad spectrum Ampicillin/Amoxicillin, and Erythromycin therapy during conservative management of pPROM before 32 weeks' gestation. We further review the histopathologic correlates to pPROM, to antibiotic treatment, and to perinatal outcome, and discuss the relationships between maternal and neonatal cytokine levels intercellular adhesion molecule, and other clinical and plasma markers regarding perinatal morbidity. The use and limitations of ultrasound and vaginally collected amniotic fluid pulmonary maturity assessment are discussed. PMID- 12889590 TI - What we have learned from an antibiotic trial in fetal fibronectin positive women. AB - Clinically silent upper genital tract microbial infections have been strongly associated with spontaneous preterm delivery, especially prior to 32 weeks gestational age. This condition represents a potentially preventable or reversible cause of preterm birth using safe, readily available, and inexpensive antibiotics. Fetal fibronectin detected in cervical and vaginal secretions between 22 to 24 weeks' gestation is a powerful predictor of subsequent risk for spontaneous preterm delivery. Evidence indicates that the detection of fetal fibronectin in cervical/vaginal secretions could represent a marker for clinically silent upper genital tract microbial infection. Thus, the National Institute of Child Health and Human Development Maternal-Fetal Medicine Units (NICHD MFMU) Network performed a randomized clinical trial to determine if antibiotic treatment (metronidazole plus erythromycin) of asymptomatic women with a positive cervical or vaginal fetal fibronectin test in early pregnancy will reduce the risk of spontaneous preterm delivery. No significant difference was observed between the antibiotic- and placebo-treated groups for spontaneous preterm delivery before 37 week's gestation (relative risk 1.17, 95% confidence interval 0.80-1.70). Therefore, use of the fetal fibronectin test to identify women for antibiotic treatment in an effort to prevent preterm birth should be discouraged, and current clinical use of the fetal fibronectin test should be limited to its negative predictive value. PMID- 12889591 TI - What we have learned about preeclampsia. AB - Preeclampsia is a multisystem disorder that complicates 6% to 8% of pregnancies, with higher rates in women with preexisting hypertension, diabetes mellitus, or previous history of preeclampsia. Recent large randomized trials, including two large trials conducted by members of the Maternal-Fetal Medicine Network, have not shown a benefit in reducing the rate of preeclampsia or perinatal outcome from the use of low-dose aspirin. Secondary analysis from these trials revealed that the onset of mild gestational hypertension or mild preeclampsia at or near term was associated with minimal to low neonatal and maternal morbidities. During review of the medical records we found considerable differences among the various centers regarding the definitions of both mild and severe preeclampsia. These differences were more prevalent in those women with pre-existing hypertension or diabetes mellitus. The majority of adverse pregnancy outcomes occurred in women who developed severe gestational hypertension-preeclampsia prior to 35 weeks' gestation and in those women with previous preeclampsia and/or pre-existing vascular disease. We also found that epidural anesthesia is safe in parturients receiving low-dose aspirin in pregnancy and in women with severe preeclampsia. PMID- 12889592 TI - What we have learned about antenatal prediction of neonatal morbidity and mortality. AB - When extremely preterm birth is anticipated, a reliable estimate of neonatal outcome is essential for the parents and health care providers who face difficult management decisions. Estimates of birth weight and gestational age are most commonly used for this purpose. The National Institute of Child Health and Human Development Maternal-Fetal Medicine Units (NICHD MFMU) Network performed an observational study of data available before delivery of infants with birth weights < 1000 g. Ultrasonographic variables (estimated fetal weight, obstetrically estimated gestational age, femur length, and biparietal diameter) and clinical variables (maternal race, antenatal care, substance abuse, medical treatment, reason for delivery, fetal gender, and presentation) were studied as predictors of intrapartum stillbirth, neonatal death, survival to 120 days after birth or to discharge, and with markers of "serious" morbidity (high-grade intraventricular hemorrhage, retinopathy of prematurity, necrotizing enterocolitis, oxygen dependence at discharge or 120 days, and seizures). Survival without serious morbidity was considered "intact." Logistic regression was used to evaluate the influence of the obstetrician's opinion of viability and willingness to perform cesarean delivery for fetal distress, birth weight, growth, gender, presentation, and ethnicity on outcomes. Fetal femur length and estimated gestational age predicted survival better than did biparietal diameter or estimated fetal weight. Antenatal ultrasound and clinical data did not distinguish those infants who would suffer serious morbidity or be considered intact. Willingness to perform cesarean delivery was associated with increased likelihood of both survival and intact survival by virtually eliminating intrapartum stillbirth and reducing neonatal mortality. However, such practice was associated with an increased chance of serious morbidity among survivors below 800 g or 26 weeks'. Although obstetricians were willing to intervene for fetal indications in most cases by 24 weeks', willingness to perform cesarean delivery was associated with twice the risk for serious morbidity in survivors at that gestational age. PMID- 12889593 TI - What we have learned about conducting randomized controlled trials in the NICHD MFMU network. AB - The National Institute of Child Health and Human Development (NICHD) created the NICHD Maternal Fetal Medicine Units Network in 1986 to conduct randomized trials and observational studies in perinatal medicine to improve adverse pregnancy and infant outcomes. From 1986 to 2002, the Network has started 16 randomized trials. Five of the trials are described, with particular attention given to difficult issues that arose, such as feasibility, sample size estimation, randomization in very high risk pregnancies, changing clinical practice, importance of the study question to the investigators, and lack of recruitment. Changes that the Network group made to their organization and methodology as a result of these issues are described, together with their application to some of the other trials proposed and conducted by the group. PMID- 12889594 TI - Quantitative fluorescence cytometric measurement of estrogen and progesterone receptors: correlation with the hormone binding assay. AB - We describe, here, a rapid flow cytometry technique for the detection and quantification of estrogen (ER) and progesterone (PgR) receptors in several human cell lines and in clinical samples obtained from breast cancer tumors. ER and PgR quantitation can be very useful in patients with breast cancer as their role in diagnosis and prognosis is well established. However ligand binding assays and immunohistochemical assays are difficult to measure heterogeneity in individual cells. On the other hand, flow cytometry is a convenient tool for quantification in individual cells. Flow cytometric results with breast cancer cell lines and clinical samples were compared to those obtained by quantitative biochemical ER and PgR performed by the standard dextran-coated charcoal biochemical assay. The latter assay is affected by the level of endogenous steroids. This is also the case in the routine measurement of ER/PgR in patient's tumor cells whereby estradiol molecules in patient's serum produced negative or low values in the biochemical assay. The mAbs used in our flow cytometric method bind to their specific ER or PgR independently of whether they are preoccupied by their ligands and they produce reliable results. With the use of beads calibrated in MESF (Molecules of Equivalent Soluble Fluorochrome) units, the ER and PgR can be measured on a per cell basis. The flow cytometric method showed a strong correlation with biochemical receptor assessments of either ER alpha (ER alphaDCC, r = 0.918, p = 0.073) or PgR (PgRDCC, r = 0.75, p = 0.001). This study demonstrates that ER alpha and PgR can be detected by flow cytometry on a per cell basis in intact cells, and can be quantitated reliably in terms of MESF without the limitations of competition with serum's estradiol molecules. PMID- 12889595 TI - High serum YKL-40 levels in patients with primary breast cancer is related to short recurrence free survival. AB - YKL-40 is a growth factor for connective tissue cells and stimulates migration of endothelial cells. YKL-40 is secreted by cancer cells, and elevated serum YKL-40 in patients with metastatic breast cancer and colorectal cancer is associated with a poorer prognosis as compared to patients with normal serum YKL-40. In the present study we evaluated the associations of preoperative serum YKL-40 in 271 patients with primary breast cancer in relation to relapse-free survival and overall survival. The median follow-up time was 5.9 years. There were 77 relapses and 69 patients died. The median serum YKL-40 concentration in the patients was 57 microg/l (range 22-688 microg/l) and significantly elevated (p < 0.0001) compared to serum YKL-40 in healthy females. Nineteen percent of the patients had high serum YKL-40 (i.e., >95 percentile of healthy females). Patients with high serum YKL-40 had shorter relapse-free interval (hazard ratio (HR) = 1.77, 95% confidence interval (CI): 1.06-2.95, p = 0.028) and overall survival (HR = 1.78, 95% CI: 1.04-3.05, p = 0.036) than patients with normal serum YKL-40. Serum YKL 40 was higher (p = 0.005) in lymph node positive patients as compared to lymph node negative patients. Multivariate analysis including lymph node status, estrogen receptor status, tumor size, age, menstrual status and serum YKL-40 showed that serum YKL-40 was an independent prognostic variable of relapse-free survival (HR = 1.73, 95% CI: 1.03-2.91, p = 0.039). Our results show that serum YKL-40 in patients with primary breast cancer at time of operation is only elevated in a small group of patients, but these patients have a shorter recurrence free interval. Further studies are required to determine the biological function of YKL-40 in breast cancer. PMID- 12889596 TI - Molecular characterization of oncostatin M-induced growth arrest of MCF-7 cells expressing a temperature-sensitive mutant of p53. AB - Our previous studies have shown that treatment of MCF-7 breast cancer cells with cytokine oncostatin M (OM) results in a growth arrest and a concurrent decrease in p53 expression. It remains to be determined whether these two important events are directly connected, as changes in p53 protein levels can lead to variable biological outcomes. In this study we have generated stable cell lines (MCF7 ptsp53) that express p53Val135 a p53 temperature-sensitive mutant. We demonstrate that overexpression of the wildtype (wt) p53 at permissive temperature in MCF7 ptsp53 cells leads to growth arrest at the G2-M phase of the cell cycle. Inhibition of endogenous p53 function with the expression of mutant p53 protein at non-permissive temperature did not affect the OM-induced G1 cell cycle arrest. Microarray studies were further carried out to identify p53- and OM-regulated genes that mediate the G2/M or G1 cell cycle arrest. We show that the expression of p21 was upregulated and expressions of cdc2, cyclin B2 and protein regulator of cytokinesis 1 (PRC1) were suppressed by overexpression of the wt p53 in MCF7 ptsp53 cells at the permissive temperature. In contrast, OM treatment caused coordinate changes of mRNA expression of several cell cycle components including c/EBPdelta, cdc20, and thymidine kinase 1 (TK1) that mainly affect G1-S phase transition. All together, our results suggest that the downregulation of p53 transcription may be involved in some other cellular changes induced by OM but it is not directly connected to the antiproliferative activity of OM per se. PMID- 12889597 TI - Pain and quality of life after surgery for breast cancer. AB - BACKGROUND: In breast cancer (BC) patients, conservative surgery (CS) followed by irradiation or immediate breast reconstruction (IBR) after modified radical mastectomy (MRM) has been proposed in the attempt to avoid the negative impact of MRM on feminine body image. Regardless of the type of operation, BC patients may feel pain even without recurrent disease with poor adjustment in terms of quality of life (QL). METHODS: We adopted a questionnaire comprising the short form of the McGill Pain questionnaire, and a previously validated questionnaire able to identify four subscales exploring physical well-being, physical autonomy, relational life and psychological well-being. The questionnaire was mailed in 1999 to a consecutive series of 757 (CS: 481 cases; MRM + IBR with skin expander: 93 cases; MRM: 183 cases) disease-free patients treated for BC between March 1995 and March 1998. RESULTS: The final analysis assessed the data relating to 529 patients who underwent axillary dissection. Pain was reported by 39.7% of women with higher incidence in patients who underwent CS than in those who underwent MRM +/- IBR, but this difference did not reach statistical significance (p = 0.07). The only statistically significant difference (p < 0.05) between the surgical groups was the pain appearance that occurred earlier in the CS patients and later in the MRM + IBR patients. No other differences were observed. The women with pain had significantly worse QL scores on all of the subscales than those without. CONCLUSION: Pain after surgery for BC distress almost one-third of patients, regardless of the type of treatment, and had a negative effect on patients' QL. The different surgical procedures may marginally influence the quantitative characteristics of pain. PMID- 12889598 TI - Vitamin D growth inhibition of breast cancer cells: gene expression patterns assessed by cDNA microarray. AB - 1,25-Dihydroxyvitamin D3 [1,25(OH)2D3], the active metabolite of vitamin D, is a potent inhibitor of breast cancer cell growth. Although it is evident that 1,25(OH)2D3 inhibits growth of both estrogen receptor alpha-positive [ER alpha(+)] and -negative [ER alpha(-)] breast cancer cells, the cellular pathways contributing to these effects remain unclear. We studied the gene expression patterns in ER alpha(+) MCF-7 and ER alpha(-) MDA MB 231 human breast cancer cells following 1,25(OH)2D3 treatment, using cDNA expression arrays. Both cell lines showed a significant induction of the 1,25(OH)2D3-dependent 24-hydroxylase gene, a marker for the actions of 1,25(OH)2D3. In MCF-7 cells, 51 genes were up regulated and 19 genes were down-regulated. The up-regulated genes encoded cell adhesion molecules, growth factors/modulators, steroid receptors/co-activators, cytokines, kinases and transcription factors. Of the up-regulated genes, 40% were implicated in cell cycle regulation and apoptosis and included cyclin G1 and cyclin I, p21-activated kinase-1 (PAK-1), p53, retinoblastoma like-2 [Rb2 (p130)], insulin-like growth factor binding protein-5 (IGFBP5) and caspases. Among the down-regulated genes were ER alpha, growth factors, cytokines and several kinases. Some of these results were confirmed by real-time PCR. In MDA MB 231 cells, 20 genes were up-regulated and 13 genes were down-regulated. Very few genes directly implicated in cell cycle regulation were up-regulated. The matrix metalloproteinases formed a major class of genes that were down-regulated in the MDA MB 231 cells. Seven genes were commonly up-regulated in both cell lines and these included transforming growth factor (TGFbeta2) and Rb2 (p130). In conclusion, the gene expression profiles of the two cell lines studied were different with a few overlapping genes suggesting that different cellular pathways might be regulated by 1,25(OH)2D3 to exert its growth inhibitory effects in ER alpha(+) and ER alpha(-) cells. PMID- 12889599 TI - Effects of neurotransmitters on the chemokinesis and chemotaxis of MDA-MB-468 human breast carcinoma cells. AB - Most patients suffering from breast carcinoma do not die due to the primary tumor but from the development of metastases. Active migration of cancer cells is a prerequisite for development of these metastases. We used time-lapse videomicroscopy and computer-assisted cell tracking of MDA-MB-468 human breast carcinoma cells, which were incorporated into a three-dimensional collagen matrix, in order to analyze the migratory activity of these cells in response to different neurotransmitters. Our results show that met-enkephalin, substance P, bombesin, dopamine, and norepinephrine have a stimulatory effect on the migration of the breast cancer cells; moreover, these cells show positive chemotaxis towards norepinephrine as was analyzed by the directionality and persistence on a single-cell basis. Gamma-aminobutyric acid (GABA) however has an inhibitory effect. Endorphin and leu-enkephalin, as well as histamin and acetylcholine, had no influence on the migratory activity of the cells. In summary, we provide evidence for a strong regulatory involvement of neurotransmitters in the regulation of breast cancer cell migration, which might provide the basis for the use of the pharmacological agonists and antagonists for the chemopreventive inhibition of metastasis development. PMID- 12889600 TI - Tumor necrosis factor-alpha induces interleukin-6 production via extracellular regulated kinase 1 activation in breast cancer cells. AB - Interleukin-6 (IL-6) and interleukin-11 (IL-11) are frequently produced by breast cancer cells. These interleukins promote osteoclast formation and may mediate osteolysis at the site of breast cancer bone metastases. Transforming growth factor-beta (TGF-beta), tumor necrosis factor-alpha (TNF-alpha) and interleukin 1beta (IL-1beta) up-regulate IL-6 and IL-11 production in a cytokine-dependent fashion in breast cancer cells, but very little is known about their intracellular signaling pathways in breast cancer cells. To study TGF-beta, TNF alpha and IL-1beta regulation of IL-6 and IL-11 production in human MDA-MB-231 breast cancer cells, we established single cell clones stably expressing dominant negative (DN) forms of the mitogen-activated protein kinases p38 (p38/AF) or ERK1 (ERK1K71R). We show here, that while basal, TGF-beta and IL-1beta induced IL-6 production was similar in parental cells and in pcDNA3 control, ERK1K71R and p38/AF clones, TNF-alpha induced IL-6 production was blunted in the ERK1K71R clones. TGF-beta and IL-1beta, but not TNF-alpha, induced IL-11 production in parental MDA-MB-231 cells. Similar findings were detected in clones stably expressing p38/AF and ERK1K71R, which did not change basal IL-11 production either. In conclusion, TNF-alpha induced IL-6 production is mediated via ERK1 activation in MDA-MB-231 cells. These observations may be helpful in designing new anti-osteolytic therapies. PMID- 12889601 TI - Expression of estrogen receptor-beta in normal mammary and tumor tissues: is it protective in breast carcinogenesis? AB - Using messenger RNA (mRNA) in situ hybridization, we investigated estrogen receptor-beta (ERbeta) mRNA levels in normal mammary, benign breast tumor (BBT), breast cancer (BC), and metastatic lymph node tissues to verify the role of ERbeta in BC development and progression. ERbeta expression was significantly decreased in BC and metastatic lymph node tissues compared with normal mammary and BBT tissues (p < 0.01). The intensity and extent of ERbeta mRNA signals were also significantly lower in BC and metastatic lymph node tissues than in the normal mammary and BBT tissues (p < 0.01). An inverse relationship was found between ERbeta mRNA level and both histologic grade (p = 0.091) and progesterone receptor expression (p = 0.052) with marginal significance, but no significant association was noted between ERbeta expression in cancer tissues and the other clinico-pathologic data. The 3-year distant relapse-free survival probability was found to be independent of ERbeta expression. Collectively, ERbeta mRNA decreases in the process of BC development, but seems to be associated with poor differentiation. PMID- 12889602 TI - Proteomic characterization of nipple aspirate fluid: identification of potential biomarkers of breast cancer. AB - Mammary ductal cells are the origin for 70-80% of breast cancers. Nipple aspirate fluid (NAF) contains proteins directly secreted by the ductal and lobular epithelium in non-lactating women. Proteomic approaches offer a largely unbiased way to evaluate NAF as a source of biomarkers and are sufficiently sensitive for analysis of small NAF volumes (10-50 microl). In this study, we initially evaluated a new process for obtaining NAF and discovered that this process resulted in a volume of NAF that was suitable for analysis in approximately 90% of subjects. Proteomic characterization of NAF identified 64 proteins. Although this list primarily includes abundant and moderately abundant NAF proteins, very few of these proteins have previously been reported in NAF. At least 15 of the NAF proteins identified have previously been reported to be altered in serum or tumor tissue from women with breast cancer, including cathepsin D and osteopontin. In summary, this study provides the first characterization of the NAF proteome and identifies several candidate proteins for future studies on breast cancer markers in NAF. PMID- 12889603 TI - A phase II study of topical ceramides for cutaneous breast cancer. AB - PURPOSE: Short chain ceramides induce tumor cell apoptosis in preclinical models. Limited therapeutic options for patients with cutaneous breast cancer prompted the testing of these sphingolipids in patients with this disease. PATIENTS AND METHODS: Twenty-five patients with refractory, cutaneous breast cancer were treated twice a day with a 1% mixture of topical C2 and C6 ceramides administered in a 1:1 ratio. For the first 8 weeks, patients were not allowed to receive other antineoplastic therapy. In addition to tumor status and toxicity assessment throughout the trial, skin biopsies for evidence of apoptosis and quality of life questionnaires (FACT-BR) were completed at baseline and 1 month. RESULTS: Only one patient manifested a partial response with topical ceramides, yielding a response rate of 4% (90% confidence interval 0, 17.6%). Median cutaneous progression-free survival was 2 months. The topical ceramides were also well tolerated, with no grade 3 or 4 toxicity reported. None of the six patients who underwent serial skin biopsies showed increased tumor cell apoptosis morphologically or by the modified TUNEL assay. CONCLUSION: To our knowledge, this trial is one of the first clinical investigations of short chain ceramides. This trial's results are not promising enough to merit further study of ceramides in the manner prescribed. PMID- 12889604 TI - Breast cancer metastasis alters acetylcholinesterase activity and the composition of enzyme forms in axillary lymph nodes. AB - Because of the probable involvement of cholinesterases (ChEs) in tumorigenesis, this research was addressed to ascertaining whether breast cancer metastasis alters the content of acetylcholinesterase (AChE) and/or butyrylcholinesterase (BuChE) in axillary lymph nodes (LN). ChE activity was assayed in nine normal (NLN) and seven metastasis-bearing nodes (MLN) from women. AChE and BuChE forms were characterised by sedimentation analyses, hydrophobic chromatography and western blotting. The origin of ChEs in LN was studied by lectin interaction. AChE activity dropped from 21.6 mU/mg (nmol of the substrate hydrolysed per minute and per milligram protein) in NLN to 3.8 mU/mg in MLN (p < 0.001), while BuChE activity (3.6 mU/mg) was little affected. NLN contained globular amphiphilic AChE dimers (G2A, 35%), monomers (G1A, 30%), hydrophilic tetramers (G4H, 8%), and asymmetric species (A4, 23%, and A8, 4%); MLN displayed only G2A (65%) and G1A (35%) AChE forms. NLN and MLN contained G4H (79%), G4A (7%), and G1H (14%) BuChE components. Neither the binding of ChE forms with lectins and antibodies nor the subunit size were altered by metastasis. The higher level of AChE in NLN than in brain and the specific pattern of AChE forms in NLN support its role in immunity. The different profile of AChE forms in NLN and MLN may be useful for diagnosis. PMID- 12889605 TI - Breast cancer diagnosis by scintimammography: a meta-analysis and review of the literature. AB - Scintimammography is a relatively new, non-invasive diagnostic modality in the evaluation of breast cancer. The purpose of the current study was to review the existing literature on the accuracy of scintimammography in the diagnosis of breast cancer. A search of all articles published between 1st January 1967 and 31st December 1999 was conducted. A total of 64 unique studies were selected. Each scientific paper was reviewed for scientific merit by an epidemiologist, a surgeon and a surgical resident. Assessment of scientific merit was based on a scoring scheme developed for the study. The articles included in this review reported data on a total of 5340 patients assessed for breast cancer with scintimammography. The aggregated summary estimates on these patients were sensitivity: 85.2% and specificity: 86.6%. For patients with a palpable mass the sensitivity and specificity were 87.8 and 87.5%, respectively. For patients without a palpable mass the sensitivity was 66.8% and that for specificity was 86.9%. The results of this review have shown that scintimammography may be used effectively as an adjunct to mammography and physical examination in the diagnosis of breast cancer. PMID- 12889606 TI - Estrogen receptor alpha gene polymorphisms and breast cancer risk. AB - We conducted a hospital-based case-control study to evaluate the association between the XbaI and PvuII restriction fragment length polymorphisms (RFLPs) in intron I of the estrogen receptor alpha (ER alpha) gene and breast cancer risk. The study population consisted of 205 histologically confirmed incident breast cancer cases and 205 age-matched controls with no present and previous history of cancer. The PvuII genotype distribution did not show any difference between cases and controls, but the adjusted odds ratio (OR) for the XbaI X allele containing genotypes was 0.4 (95% CI: 0.3-0.6) compared with the xx genotype. The decrease in the OR appeared to be more attributable to the postmenopausal women; the ORs were 0.3 (95% CI: 0.1-0.5) and 0.5 (95% CI: 0.3-0.9) for postmenopausal and premenopausal women, respectively. Our results therefore suggest that the ER alpha XbaI polymorphism modifies individual susceptibility to breast cancer in Korean women. PMID- 12889607 TI - Anaerobic desulphurisation of thiophenes by mixed microbial communities from oilfields. AB - Anaerobic enrichment cultures obtained from oil fields degraded various thiophenic compounds i.e. thiophene, benzothiophene and dibenzothiophene, with the concomitant formation of sulphide using hydrogen, lactate and ethanol as possible electron donors. It was demonstrated that dibenzothiophene was converted to biphenyl. However, hydrocarbon products from benzothiophene and thiophene desulphurisation could not be detected. After further enrichment on thiophenic compounds as the sole electron acceptor, the conversion activity disappeared while homo-acetogenic bacteria became abundantly present. In order to gain stable conversions of thiophenic compounds, attempts were made to isolate the sulphide producing bacteria. Two highly enriched cultures were obtained, which degraded thiophenic compounds, but the activity remained low and homo-acetogenesis remained dominant. PMID- 12889608 TI - Isolation and identification of thiocyanate utilizing chemolithotrophs from gold mine soils. AB - A mixed bacterial culture capable of growing in potassium-thiocyanate containing medium (200 mg KSCN) has been isolated from bacterial suspensions of soil samples collected near gold mines in Kumjung (Korea). The isolates were initially characterized by metabolic profile analysis and were identified as Bacillus thermoglucosidasius, Bacillus cereus, Bacillus licheniformis, Bacillus mycoides, Brevibacterium epidermidis, Brevibacterium otitidis, and Corynebacterium nitrilophilus. One of the seven isolates was initially characterized as Brevibacterium epidermidis, which is not known to degrade thiocyanate. However, using 16S rDNA sequencing, this strain was identified as a member of Klebsiella. The strain shows high similarity values (95.8 to 96.4%) with Klebsiella species, and the closest known relative was found to be K. ornithinolytica ATCC 31898. The result indicates that species of the genus Klebsiella were the closest phylogenetic relatives of the investigated strain. This is the first known report of a member of Klebsiella that is capable of utilizing thiocyanate as sole source of carbon and nitrogen. PMID- 12889609 TI - Sulfite-oxido-reductase is involved in the oxidation of sulfite in Desulfocapsa sulfoexigens during disproportionation of thiosulfate and elemental sulfur. AB - The enzymatic pathways of elemental sulfur and thiosulfate disproportionation were investigated using cell-free extract of Desulfocapsa sulfoexigens. Sulfite was observed to be an intermediate in the metabolism of both compounds. Two distinct pathways for the oxidation of sulfite have been identified. One pathway involves APS reductase and ATP sulfurylase and can be described as the reversion of the initial steps of the dissimilatory sulfate reduction pathway. The second pathway is the direct oxidation of sulfite to sulfate by sulfite oxidoreductase. This enzyme has not been reported from sulfate reducers before. Thiosulfate reductase, which cleaves thiosulfate into sulfite and sulfide, was only present in cell-free extract from thiosulfate disproportionating cultures. We propose that this enzyme catalyzes the first step in thiosulfate disproportionation. The initial step in sulfur disproportionation was not identified. Dissimilatory sulfite reductase was present in sulfur and thiosulfate disproportionating cultures. The metabolic function of this enzyme in relation to elemental sulfur or thiosulfate disproportionation was not identified. The presence of the uncouplers HQNO and CCCP in growing cultures had negative effects on both thiosulfate and sulfur disproportionation. CCCP totally inhibited sulfur disproportionation and reduced thiosulfate disproportionation by 80% compared to an unamended control. HQNO reduced thiosulfate disproportionation by 80% and sulfur disproportionation by 90%. PMID- 12889610 TI - Heavy metal bioavailability in a soil affected by mineral sulphides contamination following the mine spillage at Aznalcollar (Spain). AB - A field experiment, lasting 14 months, was carried out in order to assess the effect of organic amendment and lime addition on the bioavailability of heavy metals in contaminated soils. The experiment took place in a soil affected by acid, highly toxic pyritic waste from the Aznalcollar mine (Seville, Spain) in April 1998. The following treatments were applied (3 plots per treatment): cow manure, a mature compost, lime (to plots having pH < 4), and control without amendment. During the study two crops of Brassica juncea were grown, with two additions of each organic amendment. Throughout the study, the evolution of soil pH, total and available (DTPA-extractable) heavy metals content (Zn, Cu, Mn, Fe, Pb and Cd), electrical conductivity (EC), soluble sulphates and plant growth and heavy metal uptake were followed. The study indicates that: (1) soil acidification, due to the oxidation of metallic sulphides in the soil, increased heavy metal bioavailability; (2) liming succeeded in controlling the soil acidification; and (3) the organic materials generally promoted fixation of heavy metals in non-available soil fractions, with Cu bioavailability being particularly affected by the organic treatments. PMID- 12889611 TI - Performance and ethanol oxidation kinetics of a sulfate-reducing fluidized-bed reactor treating acidic metal-containing wastewater. AB - The treatment of simulated acidic wastewater (pH 2.5-5) containing sulfate (1.0 2.2 g l(-1)), zinc (15-340 mg l(-1)) and iron (57 mg l(-1)) was studied in a sulfate-reducing fluidized-bed reactor (FBR) at 35 degrees C. The original lactate feed for enrichment and maintenance of the FBR culture was replaced stepwise with ethanol over 50 days. The robustness of the process was studied by increasing stepwise the Zn, sulfate and ethanol feed concentrations and decreasing the feed pH. The following precipitation rates were obtained: 360 mg l(-1) d(-1) for Zn and 86 mg l(-1) d(-1) for Fe, with over 99.8% Zn and Fe removal, with a hydraulic retention time of 16 h. Under these conditions, 77-95% of the electrons were accepted by sulfate reduction. The alkalinity produced from ethanol oxidation increased the wastewater pH from 2.5 to 7.5-8.5. Michaelis Menten constants (Km) determined in batch FBR experiments, were 4.3-7.1 mg l(-1) and 2.7-3.5 mg l(-1) for ethanol and acetate oxidation, respectively. The maximum oxidation velocities (Vmax) were 0.19-0.22 mg gVS(-1) min(-1) and 0.033-0.035 mg gVS(-1) min(-1), for ethanol and acetate, respectively. In summary, the FBR process produced a good quality effluent as indicated by its low organic content and Zn and Fe concentrations below 0.1 mg l(-1). PMID- 12889612 TI - The role of sulphidogenesis in anaerobic treatment phase of tannery wastewater treatment in advanced integrated wastewater pond system. AB - An outdoor experiment was conducted to study the competition between Sulphate Reducing Bacteria (SRB) and Methanogenic Archaea (MA) in anaerobic treatment phase of tannery wastewater treatment in pilot-scale Advanced Facultative Pond (AFP). The relative electron flow towards sulphate reduction was higher (59-83%) than towards methanogenesis (17-41%), although the COD recovery within the reactor varied between 15 and 90%. The results also demonstrated that the flow of electrons towards SRB increased with increase of the sulphate concentration and decrease of the COD:SO4= ratio. PMID- 12889613 TI - Use of sulfate reducing cell suspension bioreactors for the treatment of SO2 rich flue gases. AB - This paper describes a novel bioscrubber concept for biological flue gas desulfurization, based on the recycling of a cell suspension of sulfite/sulfate reducing bacteria between a scrubber and a sulfite/sulfate reducing hydrogen fed bioreactor. Hydrogen metabolism in sulfite/sulfate reducing cell suspensions was investigated using batch activity tests and by operating a completely stirred tank reactor (CSTR). The maximum specific hydrogenotrophic sulfite/sulfate reduction rate increased with 10% and 300%, respectively, by crushing granular inoculum sludge and by cultivation of this sludge as cell suspension in a CSTR. Operation of a sulfite fed CSTR (hydraulic retention time 4 days; pH 7.0; sulfite loading rate 0.5-1.5 g SO3(2-) l(-1) d(-1)) with hydrogen as electron donor showed that high (up to 1.6 g l(-1)) H2S concentrations can be obtained within 10 days of operation. H2S inhibition, however, limited the sulfite reducing capacity of the CSTR. Methane production by the cell suspension disappeared within 20 days reactor operation. The outcompetition of methanogens in excess of H2 can be attributed to CO2 limitation and/or to sulfite or sulfide toxicity. The use of cell suspensions opens perspectives for monolith or packed bed reactor configurations, which have a much lower pressure drop compared to air lift reactors, to supply H2 to sulfite/sulfate reducing bioreactors. PMID- 12889614 TI - Gastric bypass for morbid obesity in patients 50 years or older: is laparoscopic technique safer? AB - Some physicians have considered age > or = 50 years as a relative contraindication for bariatric surgery. Recent reports demonstrated the safety and efficacy of Roux-en-Y gastric bypass (RYGB) in this patient subgroup, but comparisons between laparoscopic technique (LT) and open technique (OT) have not been reported. A review of 52 patients > or = 50 years old who underwent RYGB between January 1999 and April 2002 was conducted. Demographics, operative data, and outcomes were assessed. Preoperative and postoperative renal and hepatic functions, electrolytes, anemia studies, and hematology results were compared. Patients were divided into LT and OT groups and operative outcomes were compared. The percentage of excess body weight loss was 66 +/- 4 per cent at mean follow-up of 12 months. Blood samples drawn after a mean of 8 +/- 2 months revealed no postoperative metabolic alterations. RYGB resulted in a reduction of the number of patients with hyperglycemia, hypertension, degenerative joint disease, gastroesophageal reflux disease, and continuous positive airway pressure dependent sleep apnea (P < 0.05). The LT resulted in fewer intensive care unit admissions and shorter length of stay. RYGB is safe and well tolerated in patients > or = 50 years resulting in no renal, hepatic, or electrolytic alterations. Weight loss and control of obesity-related comorbidities are satisfactory. The LT results in fewer intensive care unit admissions and shorter length of stay than the OT. PMID- 12889615 TI - Current management of common bile duct stones in a teaching community hospital. AB - The advent of laparoscopic cholecystectomy (LC) has complicated management of common bile duct (CBD) stones. While LC is routine, laparoscopic CBD exploration (LCBDE) is not, and an algorithm to manage suspected choledocholithiasis has not been uniformly accepted. We evaluated current management of choledocholithiasis. Patients suspected of having CBD stones over a 2-year period were evaluated, and 42 studies in the literature were reviewed. Thirty-two patients were identified. Fourteen patients (44%) had LC with intraoperative cholangiogram (IOC) with no preoperative studies. IOC revealed CBD stones in nine (64%). Seven had CBD exploration (CBDE) at cholecystectomy, and two had postoperative endoscopic retrograde cholangiopancreatography (ERCP). CBDE was successful in five cases, and ERCP was successful in one. Eighteen patients (56%) underwent preoperative ERCP. Five (28%) had no CBD stones. ERCP removed stones in nine patients, and four had open CBDE after failed ERCP. Current literature supports LC with IOC without any preoperative studies. Laparoscopic CBDE is highly successful but depends on surgeon experience. Removing CBD stones with ERCP is also very successful but is associated with increased cost, hospital stay, and complications. We conclude that LC with IOC should be performed without preoperative ERCP when choledocholithiasis is suspected. If found, stones should be removed laparoscopically if possible. PMID- 12889616 TI - Nontherapeutic laparotomies revisited. AB - A wide variability exists in the reported incidents of complications following nontherapeutic laparotomy for trauma. We undertook this study to examine the decision-making leading to and complication rates related to the use of nontherapeutic laparotomy in an era of nonoperative management. We conducted a retrospective chart review of all nontherapeutic laparotomies as defined by the operating surgeon performed between May 1998 and May 2001. A total of 50 patients (6%) underwent nontherapeutic laparotomies predominantly for penetrating injury. The most frequent preoperative reason for nontherapeutic laparotomy was the question of hollow viscous or diaphragmatic injury on preoperative studies. Peritoneal signs on examination and peritoneal penetration on wound exploration were also frequent indications for surgery. Significant complication rates were low at approximately 12 per cent. Total length of stay was 7.3 days and 5 days if patients with significant other associative injuries were excluded. Overall morality was 4 per cent and unrelated to the nontherapeutic laparotomy in all cases. Overall incidence of nontherapeutic laparotomy has decreased with the use of abdominal helical CT and triple-contrast CT to evaluate penetrating injuries. Despite these advances diaphragmatic and hollow viscous injuries remain a concern. Although overall serious complication rates were low the use of nontherapeutic laparotomy adds significantly to length of stay and complications. PMID- 12889617 TI - Minimally invasive surgery for pediatric solid neoplasms. AB - The role of minimally invasive surgery (MIS) in children with solid neoplasms is slowly evolving. MIS appears to be an ideal way to obtain diagnostic information (i.e., tissue biopsy) in children with solid neoplasms, but its role as an ablative/curative technique is controversial. We examined the safety, reliability, and outcome of decisions made on the basis of MIS performed in children with solid neoplasms. A total of 28 children (19 boys and nine girls; age range, 14 months to 17 years) with solid neoplasms underwent 29 MIS procedures between July 1, 2000 and June 30, 2002. Complications, biopsy results, and outcomes were reviewed. Successful ablation via MIS was defined as clear microscopic margins on permanent pathology and no evidence of remnant disease on follow-up diagnostic radiological examination. There were 20 thoracoscopic and nine laparoscopic procedures. Laparoscopy included purely diagnostic without tissue biopsy or simply determination of resectability (two), incisional biopsy (two), and excisional biopsy (five; two adrenalectomy and three oophorectomy). Thoracoscopy included 15 lung biopsies and five biopsies of mediastinal masses. Diagnostic accuracy was 100 per cent in all cases. MIS as an ablative technique was successful in 10 of 10 cases. No children were found retrospectively to have been inadequately treated via MIS. We conclude that MIS can be used safely and successfully to diagnose children with suspicious solid neoplasms. Furthermore MIS may have a role as an ablative/curative technique in carefully selected circumstances. PMID- 12889618 TI - Isolated internal iliac artery aneurysm resection and reconstruction: operative planning and technical considerations. AB - Isolated iliac artery aneurysms are rare but dangerous aneurysms associated with a high incidence of rupture (between 14 and 70%). Rupture is frequently associated with an exceedingly high mortality primarily because of the elusive nature of the presenting symptoms and the resulting major delays in treatment. Accordingly these aneurysms are best managed aggressively. Although emerging endovascular techniques show promise surgical resection and reconstruction remains the gold standard for definitive management and has withstood the test of time with excellent durable and unparalleled results. That said, from an operative perspective these aneurysms are technically demanding and remain one of the more formidable technical challenges in vascular surgery. To highlight the key elements involved in a successful repair we present a right internal iliac artery aneurysm with an associated contralateral common iliac artery occlusion, review the necessary preoperative planning and the available surgical treatment options, and detail the technical steps leading to a successful reconstruction. Careful operative planning is critical. Inadequate preoperative studies, inadequate preoperative decision making, and a poorly formulated operative strategy can lead to catastrophic results. Some of the most feared complications include pelvic venous injury with resulting massive hemorrhage and postoperative pelvic ischemia (with resulting rectal and/or spinal cord ischemia) which occurs as a result of inadequate contralateral collateral pelvic blood flow when the internal iliac artery is not reimplanted. Accordingly the preoperative workup must include a careful analysis of the adequacy of the contralateral pelvic blood flow to supply collateral flow in the event that the internal iliac is not reimplanted. In the presence of compromised contralateral internal iliac perfusion, resection and reconstruction or an alternative form of pelvic revascularization is mandatory. Excellent and unencumbered exposure is mandatory for a safe and successful repair. The retroperitoneal approach as illustrated in this case is strongly recommended. Although it is challenging excellent results can be achieved by resection of the aneurysm and reconstruction. PMID- 12889619 TI - Clinical impact of micrometastasis of the lymph node in gastric cancer. AB - Micrometastasis in regional lymph nodes has been observed immunohistochemically, but the biological and clinical roles of minute nodal invasion of carcinoma in gastric cancer remain unclear. We used the anti-cytokeratin (AE1/AE3) antibody to immunohistochemically detect nodal micrometastatic lesions that could not be identified by routine pathological examination. A total of 4203 lymph nodes were examined in 180 gastric cancer patients. Lymph node metastasis was found in 36 of the 180 patients by routine pathological evaluation. Immunohistochemically micrometastasis was detected in the lymph nodes of 19 node-negative patients. Micrometastasis was not detected in any of the mucosal gastric cancer patients who underwent lymph node dissection. Gastric cancer patients with more than six metastatic lymph nodes all had nodal micrometastasis. Patients with micrometastasis had a significantly poorer survival rate than those without micrometastasis (P < 0.05). Based on the present results the presence of lymph node micrometastasis may provide a more accurate indication for surgical outcome in gastric cancer patients at the same clinical stage. PMID- 12889620 TI - Hand-assisted laparoscopic colectomy: a single-institution experience. AB - The purpose of this study was to examine the results of a single institution experience with hand-assisted laparoscopic colon resection for benign disease. We conducted a retrospective study of consecutive cases performed by experienced laparoscopic surgeons at a single institution. From August 1999 to June 2001, 37 patients underwent hand-assisted laparoscopic colon resection. Seventeen patients were male, and 20 were female. Median patient age was 58 years (range 20-80). Indications for surgery were: polyp (13), uncomplicated diverticular disease (eight), complicated diverticular disease (i.e., colovesicular fistula, phlegmon, etc.) (seven), chronic constipation (four), rectal prolapse (two), ulcerative colitis (one), endometriosis (one), and fecal incontinence (one). Procedures performed were: sigmoidectomy (14), right colectomy (nine), low anterior resection (seven), subtotal colectomy (five), cecectomy (one), and transverse colectomy (one). Variables examined were: conversion to open procedure, operative time, blood loss, time to return of flatus, length of postoperative hospital stay, and complications. There were no deaths. One case was converted to celiotomy (unable to rule out malignancy). The median operative time was 122 minutes (range 32-240) with a median operative blood loss of 132 mL (range 0 300). Return of flatus was noted (median) at postoperative day 3 (range 1-5), and the median length of stay after operation was 4 days (range 2-8). One patient developed a superficial wound infection, and there was one pelvic abscess (drained percutaneously). One patient developed urinary retention. There were no reoperations. In this single-institution experience hand-assisted laparoscopic elective colectomy for benign disease was successful in both straightforward and complicated cases. A low conversion rate to celiotomy and favorable operative times compared with published "pure" laparoscopic results suggest a flatter learning curve for handoscopy while retaining the benefits of "minimally invasive" surgery such as early return of flatus and short postoperative hospital stay. For these reasons hand-assisted laparoscopy should be considered an acceptable technique in elective colon resection for benign disease. PMID- 12889621 TI - Operative management and outcome of iliac vessel injury: a ten-year experience. AB - The purpose of this study is to review demographics and examine and stratify risk factors related to the outcome of operatively treated iliac vascular injuries. We conducted a retrospective review of 78 cases of iliac vessel injury. Patients with blunt and penetrating injury had statistically similar length of hospital stay and intensive care unit stay, incidence of shock, and mortality. Shock on admission and bleeding hematoma are linked, and shock increased mortality with an odds ratio of 5.2 (P = 0.002). A review of operative technique and outcome demonstrated a low mortality of 25 per cent in arterial bypass of an isolated arterial injury versus a mortality of 83 per cent in the combined injury group. Patients treated with primary repair of venous injuries had a lower incidence of shock and mortality compared with patients treated with venous ligation. We conclude that, if matched for severity of injury and physiologic instability, the mechanism of injury does not affect mortality. Shock is the most significant prognostic factor for mortality. Operative management must be based on presence of shock. PMID- 12889622 TI - Effect of hyperglycemia and nitric oxide synthase inhibition on heat tolerance and induction of heat shock protein 72 kDa in vivo. AB - Diabetes and nitric oxide synthase (NOS) inhibition both exacerbate mesenteric ischemia/ reperfusion injury. Heat shock protein 72 (HSP-72) protects against KDa ischemia/reperfusion damage in vivo. The effect of diabetes on HSP-72 expression in vivo is unknown. The aim of this study was to determine the effects of diabetes and NOS inhibition on HSP-72 induction in vivo. Rats were assigned to four groups: control (C), streptozotocin-induced diabetic (D), acute hyperglycemia (A), and L-N(omega)-nitro-L-arginine treated (L). Rats were subjected to hyperthermia and allowed to recover for 4 hours. Intestine and liver samples from heated (H) and nonheated (NH) rats were analyzed for HSP-72 by Western blot. HSP-72 levels were increased significantly in CH compared to CNH rats. No deaths occurred in CH rats; however, death rates were significant in AH, DH, and LH rats. DH rats died earlier than LH and AH rats. HSP-72 in liver and intestine was reduced significantly in LH rats. When compared with CH rats the surviving AH and DH rats exhibited similar HSP-72 levels in the liver. Diabetes, acute hyperglycemia, and L-N(omega)-nitro-L-arginine treatment lower heat stress tolerance. NOS is required for HSP-72 expression, but not survival. Diabetics who survive heat stress moderately express HSP-72. Characterization of altered thermotolerance and HSP-72 may provide mechanisms for the deranged diabetic stress response. PMID- 12889623 TI - [18F]Fluorodeoxyglucose positron emission tomography surveillance of hepatic metastases from prostate cancer following radiofrequency ablation: a case report. AB - The liver is the organ most commonly involved with metastatic disease. Surgical resection of hepatic metastases is the only potentially curative therapy, but it is possible in only 20 per cent of the patients. Radiofrequency ablation (RFA) of hepatic lesions is a therapeutic option for unresectable hepatic metastases. Today there is no clear consensus about which imaging technique is the most reliable to monitor RFA therapy. [18F]Fluorodeoxyglucose (FDG) positron emission tomography (PET) is a new imaging modality allowing evaluation of glucose metabolism that has become established for monitoring therapy and early detection of recurrence of various types of malignant tumors. We present a case report of a 61-year-old man treated for prostate carcinoma 3 years earlier who presented with rising serum prostate-specific antigen (PSA) levels. A CT scan demonstrated two hepatic metastases that were treated with RFA because the patient refused surgery. During 3 years of follow-up hepatic recurrence was monitored with serum PSA levels, CT of the abdomen, and FDG-PET imaging on multiple occasions. On three separate occasions FDG-PET revealed hypermetabolic foci despite no definite evidence of recurrence on CT. Furthermore FDG-PET imaging 2 months after the last RFA therapy showed two large photopenic areas without evidence of hypermetabolism consistent with successful RFA therapy. Serum PSA levels correlated better with FDG-PET than CT results. We conclude that in this patient FDG-PET imaging was more accurate than CT for monitoring recurrence of hepatic metastases from prostate carcinoma after RFA therapy. PET demonstrated hypermetabolic foci when there was recurrence and no evidence of hypermetabolism early after successful RFA therapy. In addition FDG-PET imaging helped to guide the placement of the RFA probe to the most metabolically active part of the tumor. PMID- 12889624 TI - Are you ready to become a robo-surgeon? AB - Robotic and minimally invasive surgery represents the future of modern surgical care. However, its role during the training of surgical residents has yet to be investigated. A previous study conducted by our group surveyed program directors at accredited general surgery training programs in the United States to determine the prevalence and application of robotics in their residency programs. This current study is a follow-up survey sent to residents across the United States to see whether they were being adequately trained and exposed to robotic surgery during their training. A survey was sent to 1800 general surgery residents, and their responses were tabulated and analyzed. Twenty-three per cent of the 1800 residents responded to our survey. An overwhelming 57 per cent of the responders indicated a high interest in robotic surgery. However, 80 per cent of the responders indicated not having a robotic training program. Robotic surgery has led to many promising advancements within the surgical subspecialties. With this emerging technology comes the need for a greater emphasis on the training of surgeons in robotics during their residency. PMID- 12889625 TI - Mini-laparotomy cholecystectomy in the era of laparoscopic cholecystectomy: a community-based hospital perspective. AB - Laparoscopic cholecystectomy is clearly the gold standard for symptomatic cholelithiasis. Open cholecystectomy is now reserved for difficult and problematic cases. The purpose of this paper is to propose that mini-laparotomy cholecystectomy (minicholecystectomy) can be as effective as laparoscopic cholecystectomy. This paper compares the two techniques in well-matched patients. In addition mastery of this technique is practical and rewarding and should be part of the repertoire of the general surgeon. We conducted a retrospective review of the experience of a single surgeon at a community-based teaching hospital over a 2-year period for minicholecystectomy and laparoscopic cholecystectomy. Sixty-six patients were matched for age, sex, body surface area, and Acute Physiology and Chronic Health Evaluation II score. The absolute cost was lower for the minicholecystectomy group than for the laparoscopic cholecystectomy group. The operating room times were not significantly different in the two groups (P value 0.79). The average length of stay and the average amount of intramuscular analgesia required for the two groups were also not significantly different (P values 0.69 and 0.35, respectively). Although subjective postoperative satisfaction was equal for both groups the minicholecystectomy group had no complications whereas the laparoscopic group had two (myocardial infarction and cystic duct stump leak) complications. We conclude that minicholecystectomy can be used as a viable alternative to laparoscopic cholecystectomy especially in patients who cannot tolerate laparoscopic procedures and in areas where cost containment is critical. PMID- 12889626 TI - Management of adult splenic injury: a 20-year perspective. AB - The objective of this study was to identify changes in the management of blunt splenic injury in adults. Hospital discharge abstract data from Maine were used to identify patients > or = 16 years old discharged between January 1, 1981 and December 31, 2000 with International Classification of Diseases (9th revision) codes indicating splenic injury. Incidence of operative intervention and outcomes for trauma hospitals and other hospitals were determined. The number of splenic injuries was about 75 per year for the first 15 years increasing to 96 per year in the last 5-year period. The rate of operative intervention declined from 71 to 41 per cent and was lower at the trauma hospitals during the last two 5-year periods (41% vs 53%). The rate of operative intervention > 24 hours after admission was 2.2 per cent during the last 10 years of study. The rate of splenorrhaphy remained stable at about 12 per cent throughout the study period. Mortality rates were higher at the trauma hospitals as were the numbers of patients with very severe injuries (Injury Severity Score > 25). There has been a marked decrease in rate of operative intervention for splenic injury in adults especially in the last 10 years. Rates of operative intervention were lower at the trauma hospitals despite higher injury severity. Frequency of delayed intervention was low and did not increase with lower operative rate. PMID- 12889627 TI - Multimodality management of locally advanced rectal cancer. AB - Despite the routine use of adjuvant chemoradiation for curatively resected stage II and III rectal cancer a significant percentage of patients ultimately fail locally and/or distally; this underscores the need for continued improvement in the efficacy of combined-modality therapy and quality of rectal cancer resection. The recognition of the significance of lateral or circumferential margins of resection has paralleled the widespread use of total mesorectal excision. In addition to facilitating negative margins of resection and local control, sharp mesorectal techniques also facilitate identification and preservation of pelvic autonomic nerves thereby greatly reducing the incidence of urinary and sexual dysfunction following radical resection. Lastly, restorative options can result in excellent bowel function in carefully selected patients undergoing a "very low" anterior resection. Efforts are currently directed at identifying the subset of locally advanced rectal cancer patients who may be adequately treated with a resection alone thereby avoiding the added morbidity of adjuvant radiation and chemotherapy. PMID- 12889628 TI - Recurrent eosinophilic mastitis. AB - Systemic diseases rarely present with manifestations in the breast. Disorders such as sarcoidosis and pyoderma gangrenosum have been described to produce signs and symptoms in the breast. Peripheral eosinophilia is a systemic disease associated with a finite group of conditions including asthma, allergic or atopic disease, collagen vascular disease, and parasitic infection. In addition it has been described in association with several malignancies. A 50-year-old woman with a history of asthma and significant eosinophilia presented with an enlarging breast mass. Complete excision of the mass revealed eosinophilic mastitis. Two years later the patient again presented with a large mass in the same breast. Repeat excision demonstrated the same pathology. The clinical significance of this lesion is important because it presents as an enlarging, painless, dominant mass. Excision is necessary for management and to conclusively rule out malignancy. Its recurrence despite excision to negative margins, however, may indicate that control of the eosinophilia--and possibly the underlying disorder- is important to prevent further recurrence. PMID- 12889629 TI - Robotic-assisted minimally invasive transhiatal esophagectomy. AB - Minimally invasive surgery has been increasing in its adaptability to a wide range of procedures. Initially used in general surgery for cholecystectomy its use has now expanded to include Nissen fundoplications, Heller myotomies, donor nephrectomies, and total esophagectomies. Technological advancements have evolved to include robotic systems for performance of complex surgical procedures. We report on our experience of using robotic-assisted technology to perform a transhiatal total esophagectomy. PMID- 12889630 TI - The Mesopotamian schools of Edessa and Jundi-Shapur: the roots of modern medical schools. PMID- 12889631 TI - Re: Minimally invasive breast surgery (evolution, expansion, and what's on the horizon). PMID- 12889632 TI - Re: Surgical removal of metastatic phylodes tumor: this should be strongly considered for the solitary lesion as long as cosmetic and functional disability is not excessive. PMID- 12889633 TI - Evaluation of ground water denitrification at a biosolids disposal site. AB - A study was conducted at a sanitary sewage sludge (biosolids) disposal site in Springfield, Illinois, U.S.A. to determine if biological denitrification played a significant factor in attenuation of ground water nitrate values. The site selected for this study is a 23 ha (57 acre) dedicated biosolids disposal facility located adjacent to a 75.7 million liter per day (20 million gallons per day) municipal treatment plant that uses anaerobic solids stabilization for treatment of generated biosolids material. Biosolids have been disposed of by fixed-point spray applicators at the site since 1976, which has caused ground water nitrate levels to increase significantly above background levels. A method was developed using a conservative chemical tracer to simulate the biosolids application process and monitor the ground water directly beneath the simulated disposal site. Results demonstrated a net decline of nitrates that could not be attributed to dilution alone. While the monitoring methodology developed for this study did not directly estimate the denitrification rate, a rate for overall nitrate reduction was calculated that could be considered to take into account all transport and reduction mechanisms such as denitrification, advection, dispersion and dilution. PMID- 12889634 TI - Monitoring and assessment of land use status by GIS. AB - The objectives of this study were to assess land suitability and to monitor the spatial and temporal changes in land use types (LUTs) by using GIS. The study was conducted in an irrigated area of 920 ha. located in the East Mediterranean region of Turkey. Archived data of LUTs were utilized. Data analysis was conducted by using ArcInfo and ArcView GIS softwares. The land characteristics were compared with land requirements of present LUTs by taking into account the land evaluation results. Three major soil series and twelve different LUTs were identified. Corn and citrus were the most planted LUTs in the assessment period. Generated maps demonstrated how LUTs changed spatially and temporally. Most LUTs were clustered in certain areas indicating that farmer's selection of LUTs was affected by their habits and neighbor's behavior. Land suitability assessment results indicated that Canakci and Mursel soil series were found to be highly suitable for all LUTs while Arikli soil series was determined to be moderately suitable citrus, vegetables and watermelon. Citrus plantation maps overlaid on soil series maps demonstrated that the citrus has not been planted completely on suitable areas. Land suitability assessment showed that citrus plantation in Arikli series would result in 40% yield loss and 58% of land is not used at its potential. PMID- 12889635 TI - Bird skins in museum collections: are they suitable as indicators of environmental metal load after conservation procedures? AB - To find out whether modern conservation treatments alter the level of metals in feathers, we analysed the content of 10 metals in feathers before and after skins were washed with detergent and treated with Eulan U-33 (a commonly used preservative at museums). Feathers of 31 birds of Goshawk Accipiter gentilis, Eagle Owl Bubo bubo. Feral Pigeon Columba livia domest, and Common Eider Somateria mollissima were analysed. We found that in most cases metals were partly washed out of the feathers, but the effects were related to species and type of feather. The value of bird skins as indicators of environmental metal load is therefore affected by this treatment. It is recommended that the conservation techniques used at museums should be reconsidered if skins are intended for specimen banking for future reference in environmental monitoring schemes and research. PMID- 12889636 TI - Monitoring of diazinon concentrations and loadings, and identification of geographic origins consequent to stormwater runoff from orchards in the Sacramento River watershed, U.S.A. AB - Diazinon contamination of California's rivers has resulted in placing several rivers on the federal Clean Water Act 303d list of impaired waterways. Impaired water body listing requires the development of Total Maximum Daily Loads (TMDL). Previous studies identified stormwater related diazinon pulses in California rivers. This study was conducted to monitor diazinon concentrations in the Sacramento River watershed after rainfall events, to ascertain whether pulses could be identified and, if concentrations of concern were observed, to estimate loadings and determine geographic origins of the insecticide. TMDL development requires knowledge of contaminant sources, loadings, and geographic origins. Flow and diazinon concentrations peaked in the Sacramento River at Sacramento after the three largest storms during January and February 1994. Diazinon concentrations peaked consequent to each of three storms. Diazinon concentrations measured in the Sacramento River at Sacramento exceeded the California Department of Fish and Game acute and chronic criteria for protection of aquatic life during January and February for nine and nineteen days, respectively. Multiple exceedances were observed throughout the watershed. Diazinon loading and geographic origin differed with each of the three storms. The design of this study provides a useful template for others attempting to identify loadings and sources of contaminants in surface waters and to rectify aquatic ecosystem contamination from various land use practices. PMID- 12889637 TI - Atmospheric PAH concentrations in fine and coarse particles. AB - Measurements of the urban air concentrations of PAHs associated with PM2.5-fine and PM 10-coarse particles in Chicago on the campus of IIT were achieved using a Universal Air Sampler. Short sampling time (12 hr) and high flow rates were used to measure the PAH concentrations in fine and coarse particles. Measured ambient concentrations of PAHs were classified based on wind direction and back trajectory calculations as Land and Lake samples. Differences in ambient concentrations of PAHs were observed between Land and Lake samples. Fine particle concentrations varied from 9.5 to 25.7 ng m(-3) and averaged 18.2 ng m(-3) for the Land samples, while they ranged from 4.2 to 31.5 ng m(-3) and averaged 13.4 ng m(-3) for the Lake samples. The measured PAH concentrations in coarse particles varied from 6.2 to 22.1 ng m(-3) and averaged 12.9 ng m(-3) for the Land samples, and they ranged from 2.4 to 13.0 ng m(-3) with an average value of 7.3 ng m(-3) for the Lake samples. The fine/coarse ratio of each individual PAH compound varied between 1.3 and 2.7 for the Land samples: it varied between 1.6 and 4.2 for the Lake samples. There was an increase in the fine/coarse ratio of PAH as molecular weight of the compound increases for both Land and Lake samples. PMID- 12889638 TI - Soil properties and crop productivity as influenced by flyash incorporation in soil. AB - Field experiments were carried out during 1996-97 at Gulawathi, Muthiani and Salarpur Villages, IARI Farm, New Delhi and NCPP Campus, Dadri to evaluate changes in soil characteristics and growth of wheat (Triticum aestivum L.), mustard (Brassica juncea L.), lentil (Lence esculenta Moench.), rice (Oryza sativa L.) and maize (Zea mays L.) by varying amounts of flyash addition (up to 50t ha(-1)) in soils at sowing/transplanting time of crops. Flyash addition in areas adjoining NCPP Thermal Power Plant, Dadri, Ghaziabad, U.P. ranged from 5-12 t ha(-1)] yr(-1) in 1995-96. Shoot and root growth and yield of test crops at different locations after flyash incorporation resulted in beneficial effects of flyash addition in most cases. The silt dominant texture of flyash improved loamy sand to sandy loam textures of the surface soils at the farmers' fields. The increased growth in yield of crops with flyash incorporation was possibly due to modifications in soil moisture retention and transmission characteristics, bulk density, physico-chemical characters such as pH and EC and organic carbon content. The response of flyash addition in the soil on soil health and crop productivity needs to be evaluated on long-term sustainable aspects. PMID- 12889639 TI - The marketing of health products, with special reference to do-it-yourself tests. AB - At a rapid pace, in vitro diagnostic medical devices are being developed to detect risk factors for disease; to a growing extent, these test kits are also directly marketed to the public for use at home. While at first sight serving a good cause, self testing may have considerable drawbacks relating to the well being of the test subject on the one hand, and to the privacy and the right not to know of other persons on the other. This raises the question whether the availability of self tests should be restricted. Taking into account legal developments in The Netherlands and the European Union, the author argues in favour of restrictions, at least when serious diseases or conditions are concerned. The existence of the internet is not an excuse for not adopting such safeguards. PMID- 12889640 TI - Diabetes, hypertension and birth injuries: a complex interrelationship. AB - Seemingly coincidental occurrence of various pathological conditions may derive from common etiologic denominators. While reviewing 240 malpractice claims involving shoulder dystocia related fetal injuries, we found two antenatal complications in the background conspicuously often. Chronic or pregnancy induced hypertension was identifiable in 80 instances (33%). Pregnancy induced or preexisting diabetes was diagnosed 48 times (20%). Many of these patients were poorly controlled. The blood pressure was usually checked during the antenatal visits. However, about one-half of all patients received no diabetic screening. Therefore, this study may underestimate the actual incidence of diabetes. It has been calculated that the frequency of diabetes in pregnancy and that of hypertension, is about 5% in the United States. Thus, the rates of these complications in this selected group of gravidas was severalfold higher than in the general population. Since hypertension causes retarded fetal growth, it cannot be a direct cause of arrest of the shoulders at delivery. The likely common denominator is maternal diabetes a known predisposing factor both for preeclampsia and shoulder dystocia at birth. In the course of litigations for fetal injuries, demonstration of the predisposing role of seemingly unrelated shortcomings of the medical management may profoundly influence the outcome. This principle is demonstrated by the presentation of an actual malpractice action which resulted in a substantial settlement. PMID- 12889641 TI - Regulating clinical practice: epistemological and cognitive perspectives. AB - This article analyzes the nature of clinical decision-making in order to clarify some of the impediments to the direct regulation of medical work. It develops the familiar idea of medicine as an art, showing that this includes the notion that each clinical case is unique, that clinical medical knowledge is unavoidably segmented, that clinicians commonly reason from exemplary cases, and that in this process the fine judgment of each practitioner is indispensable. Taken together this means that outside attempts to regulate medicine directly are doomed to fail. A more promising strategy would be to adopt forms of reflexive regulation which mobilize the self-regulatory capacities of medical professionals themselves. PMID- 12889642 TI - Liability in relation to the use of professional medical guidelines. AB - The author discusses the concept of professional guidelines in medicine and their relationship to potential malpractice accusations. He cites a case of omission to give a patient prophylactic medicine against post-operative thrombosis, as previously recommended in a clinical guideline. This paper develops the discussion of the problems of professional autonomy in the light of standards of medical care being increasingly governed by clinical guidelines. The question of legal liability of medical practitioners in these situations is explored. PMID- 12889643 TI - Legislative recognition in France of psychological harassment at work. AB - The recent French Law on Social Modernisation of 17 January 2002 introduced into the French Labour Code and into the French Criminal Code, the concept of "moral" harassment. The definition of psychological harassment under this law adopts quite a broad conception of the notion of psychological harassment. The legislator has established a means for "friendly" settlement of disputes: mediation. When it has not been possible to settle the dispute internally, the Courts have a number of sanctions available to them. The French Labour Code provides that any termination of the contract of employment resulting from a situation of psychological harassment is automatically null and void. Such nullification should therefore be applicable whatever the nature of the termination: dismissal, resignation or negotiated departure and it punishes psychological harassment at work by imprisonment for one year and a fine of 3,750 Euros. The French Criminal Code prescribes penalties of one year and 15,000 Euros. PMID- 12889644 TI - Ethical codes: between theory and health care practice. AB - Apart from legal norms, medicine is also governed by a broad array of ethical norms. Historically, ethical codes are a translation of these ethical norms. They are designed to assist the individual professional in the ethical aspects of his or her professional life. Ethical codes are however subject to different critiques. This contribution focuses on two critiques: (1) ethical codes neglect the pervasiveness of morality and the complexity of moral issues by presenting a set of specific ethical norms that need to be applied (outside in perspective); (2) ethical codes do not take into account the fragmentation of ethics and the decreasing homogeneity of the professional group (middle ground position). PMID- 12889645 TI - The concept of severe mental disorder and the amended law of reduced punishment for murder. AB - According to the 1995 amendment 300a of the Israeli Penal Law 1977, punishment for murder may be reduced in cases where "a severe mental disorder" is present. As a consequence, psychiatrists are called upon to define the mental disorder and to assess its impact on the actions of the murderer. This constitutes a major challenge both for the courts and for forensic psychiatrists who are still in the process of learning the ramifications of this medico-legal issue. We suggest that, from the psychiatrist's point of view, the law be used only for people suffering from mental disorders which are very closely associated with psychotic mental illness. PMID- 12889646 TI - Abortion under Greek law: parental consent for a minor's abortion. AB - As medical abortion becomes more widely used and available in Greece, several issues are emerging and require clarification. Health care providers often face a dilemma when dealing with an adolescent requesting abortion. Parents' consent is mandatory before delivering any kind of treatment to minors. However, as it appears in the case presented here, the circumstances are not always straightforward. A critical review of the Greek legal framework in relation to the current social context is attempted through an interesting case aiming to elicit potential defects of the law that should be addressed by the legislator. PMID- 12889647 TI - An ethical imperative: genetics education for physicians and patients. AB - The Human Genome project will result in a rapid increase in information about genetically based disease and possible genetically based therapies and cures. In light of this, radical changes in medical education may be necessary. Patients rely on physicians for medical advice about genetic ills and treatments and yet there is mounting evidence that physicians are not well informed about genetics and indeed that they are misinformed. There is also evidence that informed consent is not always honored in the context of genetic testing and indeed that coercion to test and even to act on test information is a practice. Misinformation and the mishandling of genetic information can cause physical, psychological and social harm to persons and intentional and non-intentional violations of informed consent, confidentiality, and privacy are causes for alarm. It is an ethical imperative that genetic literacy and education for physicians be fully and quickly addressed. PMID- 12889648 TI - Law and geriatrics: an Israeli perspective on future challenges. AB - This paper describes current problems in Israel in the fields of geriatric medicine and elder law. The background to the problems is discussed, with a commentary on the deficiencies in everyday practice. The author presents proposals for change both in the legislative system and in application of acceptable standards of medical care and practical use of improved legal provision. PMID- 12889649 TI - Health Science students' attitude to dying. AB - Health care professionals must be prepared, both as scientists and as humans, to treat, accompany and help anyone in the difficult moments prior to their death. The following study aims to identify the attitude of Health Sciences students to their own death process and the circumstances surrounding it, so as to be able to give them appropriate training in caring for those in this situation. 666 Health Sciences students at the University of Murcia were given a questionnaire dealing with different aspects of what would cause them peace or anxiety during their death process. We can see from the results that the thing that would most help them to die peacefully would be knowing that their life would not be prolonged artificially, and what worries them most is pain and suffering. We have reached the following conclusions from these results: we must train future health care professionals on a personal and professional level so that they are able to provide quality care and comfort in those situations and aspects that are associated with death in order to preclude wanting a quick death. PMID- 12889650 TI - Compulsory alcoholism treatment in New South Wales. AB - The compulsory treatment of people with substance use disorders continues to attract interest in many countries, particularly in the medico-legal, policy and research communities. Beyond the current fashion for Drugs Courts within the criminal justice system, several jurisdictions have for many years had involuntary commitment regimes operating within the civil sphere. This paper offers a case-study of one such regime that operates in New South Wales, Australia. After backgrounding the introduction of the Inebriates Act in the early 1900s, and describing the major features of the legislation, the paper outlines how the Act is currently being used, and what little is known about treatment outcomes under this regime. The paper concludes by considering a number of criticisms that have been levelled against the Inebriates Act, and briefly assessing its prospects for the future. PMID- 12889651 TI - The role of forensic experts in civil and criminal procedures in Hungary. AB - This short paper examines developments in expert evidentiary procedures in forensic cases in Hungary. The basis of current practice is described and certain problems arising from this are discussed. PMID- 12889652 TI - Confidentiality and driver licensing authorities. PMID- 12889653 TI - The professional and legal constraints on nurses to provide care. PMID- 12889655 TI - Mass spectrometric analysis of glycans in elucidating the pathogenesis of CDG type IIx . AB - The majority of secreted or membrane-bound proteins are glycosylated. The glycans attached to glycoproteins can affect a range of physicochemical and biological properties of the glycoprotein and appropriate glycosylation is essential for many normal cellular functions, with aberrant glycosylation often leading to disease. This short review briefly outlines the methodology used to release glycans from proteins and analyse them by mass spectrometry. The technology is illustrated by the description of a rapid and sensitive method for profiling glycoproteins of patients with congenital disorders of glycosylation type II. This methodology can rapidly pinpoint the defective step(s) in the processing pathway of N-linked glycans, thereby focusing the biochemical analyses that need to be performed to define the genetic basis of these diseases. PMID- 12889654 TI - Komrower Lecture. Congenital disorders of glycosylation (CDG): it's all in it! AB - Congenital disorders of glycosylation (CDGs) are due to defects in the synthesis of the glycan moiety of glycoproteins or other glycoconjugates. This review is devoted mainly to the clinical aspects of protein glycosylation defects. There are two main types of protein glycosylation: N-glycosylation and O-glycosylation. N-glycosylation generally consists of an assembly pathway (in cytosol and endoplasmic reticulum) and a processing pathway (in endoplasmic reticulum and Golgi). O-glycosylation lacks a processing pathway but is otherwise more complex. Sixteen disease-causing defects are known in protein glycosylation: 12 in N glycosylation and four in O-glycosylation. The N-glycosylation defects comprise eight assembly defects (CDG-I) designated CDG-Ia to CDG-Ih, and four processing defects (CDG-II) designated CDG-IIa to CDG-IId. By far the most frequent is CDG Ia (phosphomannomutase-2 deficiency). It affects the nervous system and many other organs. Its clinical expression varies from extremely severe to very mild (and thus probably underdiagnosed). The most interesting disease in this group is CDG-Ib (phosphomannose isomerase deficiency) because it is so far the only efficiently treatable CDG (mannose treatment). It has a hepatic-intestinal presentation. The O-glycosylation defects comprise two O-xylosylglycan defects (a progeroid variant of Ehlers-Danlos syndrome and the multiple exostoses syndrome) and two O-mannosylglycan defects (Walker-Warburg syndrome and muscle-eye-brain disease). All known CDGs have a recessive inheritance except for multiple exostoses syndrome, which is dominantly inherited. There is a rapidly growing group of putative CDGs with a large spectrum of clinical presentations (CDG-x). Serum transferrin iso-electrofocusing remains the cornerstone of the screening for N-glycosylation defects associated with sialic acid deficiency. Abnormal patterns can be grouped in to type 1 and type 2. However, a normal pattern does not exclude these defects. Screening for the other CDGs is much more difficult, particularly when the defect is organ- or system-restricted. The latter group promises to become an important new chapter in CDG. It is concluded that CDGs will eventually cover the whole clinical spectrum of paediatric and adult disease manifestations. PMID- 12889656 TI - Diagnosis of inherited disorders of liver metabolism. AB - Diagnosis of the metabolic disorder responsible for liver disease can sometimes be straightforward but it can also present a major challenge, particularly if the liver is sufficiently damaged to produce secondary biochemical abnormalities such as galactosuria, hypoglycaemia with hypoketonaemia, or excretion of 3-oxo-delta4 bile acids. It is important to consider the age of the patient, the nature of the liver disease, any extrahepatic clinical features, the imaging and the first-line laboratory tests when prioritizing diagnostic investigations. This article gives some examples of diagnoses made in our unit for patients with liver disease presenting in utero, in the neonatal period, in infancy and the preschool years, and in the school years. The differential diagnoses that should be considered for different clinical presentations are discussed. PMID- 12889658 TI - Treatment of carnitine deficiency. AB - Carnitine deficiency is a secondary complication of many inborn errors of metabolism. Pharmacological treatment with carnitine not only corrects the deficiency, it facilitates removal of accumulating toxic acyl intermediates and the generation of mitochondrial free coenzyme A (CoA). The United States Food and Drug Administration (US FDA) approved the use of carnitine for the treatment of inborn errors of metabolism in 1992. This approval was based on retrospective chart analysis of 90 patients, with 18 in the untreated cohort and 72 in the treated cohort. Efficacy was evaluated on the basis of clinical and biochemical findings. Compelling data included increased excretion of disease-specific acylcarnitine derivatives in a dose-response relationship, decreased levels of metabolites in the blood, and improved clinical status with decreased hospitalization frequency, improved growth and significantly lower mortality rates as compared to historical controls. Complications of carnitine treatment were few, with gastrointestinal disturbances and odour being the most frequent. No laboratory or clinical safety issues were identified. Intravenous carnitine preparations were also approved for treatment of secondary carnitine deficiency. Since only 25% of enteral carnitine is absorbed and gastrointestinal tolerance of high doses is poor, parenteral carnitine treatment is an appealing alternative therapeutic approach. In 7 patients treated long term with high-dose weekly to daily venous boluses of parenteral carnitine through a subcutaneous venous port, benefits included decreased frequency of decompensations, improved growth, improved muscle strength and decreased reliance on medical foods with liberalization of protein intake. Port infections were the most troubling complication. Theoretical concerns continue to be voiced that carnitine might result in fatal arrhythmias in patients with long-chain fat metabolism defects. No published clinical studies substantiate these concerns. Carnitine treatment of inborn errors of metabolism is a safe and integral part of the treatment regime for these disorders. PMID- 12889657 TI - Carnitine transport: pathophysiology and metabolism of known molecular defects. AB - Early-onset dilatative and/or hypertrophic cardiomyopathy with episodic hypoglycaemic coma and very low serum and tissue concentrations of carnitine should alert the clinician to the probability of the plasmalemmal high-affinity carnitine transporter defect. The diagnosis can be established by demonstration of impaired carnitine uptake in cultured skin fibroblasts or lymphoblasts and confirmed by mutation analysis of the human OCTN2 gene in the affected child and obligate heterozygote parents. The institution of high-dose oral carnitine supplementation reverses the pathology in this otherwise lethal autosomal recessive disease of childhood, and carnitine therapy from birth in prospectively screened siblings may altogether prevent the development of the clinical phenotype. Heterozygotes may be at risk for cardiomyopathy in later adult life, particularly in the presence of additional risk factors such as hypertension and competitive pharmacological agents. OCTN2 belongs to a family of organic cation/carnitine transporters that function primarily in the elimination of cationic drugs and other xenobiotics in kidney, intestine, liver and placenta. The high- and low-affinity human carnitine transporters, OCTN2 and OCTN1, are multifunctional polyspecific organic cation transporters; therefore, defects in these transporters may have widespread implications for the absorption and/or elimination of a number of key pharmacological agents such as cephalosporins, verapamil, quinidine and valproic acid. A third organic/cation carnitine transporter with high specificity for carnitine, Octn3, has been cloned in mice. The juvenile visceral steatosis (jvs) mouse serves as an excellent clinical, biochemical and molecular model for the high-affinity carnitine transporter OCTN2 defect and is due to a spontaneous point mutation in the murine Octn2 gene on mouse chromosome 11, which is syntenic to the human locus at 5q31 that harbours the human OCTN2 gene. PMID- 12889659 TI - L-carnitine in inborn errors of metabolism: what is the evidence? AB - A questionnaire was posted on the electronic mailing-list Metab-1 to determine current practice as regards the use of oral L-carnitine in medium-chain acyl-CoA dehydrogenase (MCAD) deficiency, propionic acidaemia (PA) and methylmalonic acidaemia (MMA). Thirty-one centres replied: L-carnitine was used routinely by 94% of respondents in PA and MMA but by only 36% in MCAD deficiency. A search was made for published papers on the use of L-carnitine in organic acidaemias and in MCAD deficiency. The quality of evidence to support the use of L-carnitine was graded according to the scale published by the Scottish Intercollegiate Guideline Network. No high-quality evidence was identified. PMID- 12889660 TI - Mitochondrial oxidative phosphorylation: pitfalls and tips in measuring and interpreting enzyme activities. AB - Mitochondrial oxidative phosphorylation (OXPHOS) is fundamental in all aspects of cellular life in aerobic cells and organisms. It is therefore not surprising that a variety of diseases have been attributed to dysfunction of the OXPHOS enzymes. Assessment of OXPHOS in human samples has proved to be a difficult task over years, even when relying on well-established methods. The complexity and the flexibility of the mitochondrial organization in cells account for a large part in the difficulties encountered in assessing OXPHOS activity. Nevertheless, a careful and detailed analysis of OXPHOS enzyme activity in cells or biopsy samples from patients at risk provides diagnosis of potential OXPHOS deficiency. Problems inherent in the use of human material, mostly the small size of the samples to be analysed, are difficult to resolve. However, cautious handling of these samples permits reasonable confidence to be reached in the interpretation of the data. PMID- 12889661 TI - Mitochondrial disorders: clinical presentation and diagnostic dilemmas. AB - The number of genes known to be involved in mitochondrial energy production and the elucidation of the function of their individual transcripts is still increasing. Although at this stage it is impossible to predict the number of human genes necessary for mitochondrial biogenesis and maintenance, the total number in humans will most probably exceed the number of mitochondrial genes found in, for example, the budding yeast, which is about 800. Without doubt we have only seen the tip of the iceberg of the clinical spectrum of mitochondrial disorders. Recent findings such as mutations in structural complex II genes in certain tumours emphasize the need to think outside the classical clinical presentation. We propose the consideration of a mitochondrial disorder in every chronic, intermittent or progressive disorder with single system or multisystem involvement, even if lactic acid is normal, and discuss such dilemmas as whether we should 'scrape the barrel' in every patient that are raised by this statement. The characterization of mitochondrial and nuclear DNA mutations in patients with enzymatically established mitochondrial defects has taught us that several of the current clinical and diagnostic assumptions have to be altered or even eliminated. The most challenging future task will be the development of new diagnostic criteria covering the expanding clinical spectrum of mitochondrial disorders. PMID- 12889662 TI - A review of primary hereditary optic neuropathies. AB - The primary inherited optic neuropathies are a heterogeneous group of disorders that result in loss of retinal ganglion cells, leading to the clinical appearance of optic atrophy. They affect between 1:10,000 to 1:50,000 people. The main clinical features are a reduction in visual acuity, colour vision abnormalities, centro-caecal visual field defects and pallor of the optic nerve head. Electrophysiological testing shows a normal flash electroretinogram, absent or delayed pattern visually evoked potentials suggestive of a conduction deficit and N95 waveform reduction on the pattern electroretinogram, consistent with a primary ganglion cell pathology. The primary inherited optic neuropathies may be sporadic or familial. The mode of inheritance may be autosomal dominant, autosomal recessive, X-linked recessive or mitochondrial. Within each of these groups, the phenotypic characteristics vary in such features as the mode and age of onset, the severity of the visual loss, the colour deficit and the overall prognosis. A number of different genes (most as yet unidentified) in both nuclear and mitochondrial genomes, underlie these disorders. The elucidation of the role of the encoded proteins will improve our understanding of basic mechanisms of ganglion cell development, physiology and metabolism and further our understanding of the pathophysiology of optic nerve disease. It will also improve diagnosis, counselling and management of patients, and eventually lead to the development of new therapeutic modalities. PMID- 12889663 TI - The eye as a window to inborn errors of metabolism. AB - Ocular manifestations in inborn errors of metabolism occur in many diseases and may be associated with any part of all eye components. In a minority of diseases it is possible to attribute the eye symptoms to a single hereditary pathogenetic mechanism. More often the aetiological relationship of the ocular defects to the metabolic disease is unknown. Diverse pathogenetic mechanisms may act via a common pathological pathway inducing ocular damage. The occurrence of eye abnormalities in metabolic disorders suggests that they are associated with direct toxic actions, errors of synthetic pathways or deficient energy metabolism. In this review, metabolic disorders with major abnormalities in the cornea, lens, retina and optic nerve are presented. In all cases, an appropriate combined approach by the ophthalmologist, paediatrician/neurologist, geneticist and clinical biochemist is the only way to diagnostic success. PMID- 12889664 TI - Overview of inherited metabolic disorders causing cardiovascular disease. AB - Inherited metabolic disorders contribute importantly to adverse cardiovascular outcomes and affect all tissue types. This review summarizes some of the more important aspects. In the venous system, heterozygosities for the factor V Leiden and prothrombin 20210G > A mutations are common and occur in 4% and 1%, respectively, of caucasians. They confer a 2- to 3- fold increase in risk of venous, but not arterial, thrombosis. Marfan syndrome affects the systemic circulation and has a population prevalence of about 1 in 4000. The more than 200 mutations responsible are in the fibrillin-1 gene (15q21.1) and mediate the characteristic skeletal, lens and aortic changes. There are two potentially lethal inherited disorders of cardiac conduction, the long QT and Brugada syndromes. The prevalence for each is about 1 in 10,000. On the other hand, autosomal dominant hypertrophic cardiomyopathies are relatively common, at 1 in 500, but with variable penetrance. Mutations are in the sarcomere proteins and more than 140 are known. Hypertrophic cardiomyopathy may be confused with Fabry disease, for which effective treatment is now available. Mutations in several genes have been shown to produce dilated cardiomyopathy in the young, but there is as yet no specific treatment. In fatty acid oxidation disorders, arrhythmias and cardiomyopathy occur during acute decompensation. An important recently established cause of cardiomyopathy is carnitine transporter defect; it is treated effectively with oral carnitine. The autosomal dominant arrhythmogenic right ventricular dysplasia occurs with a prevalence of about 1 in 15,000 and presents with arrythmias and a dilated right ventricle. The mutations responsible have been mapped to chromosomes 1, 2, 10 and 14. Lysosomal storage disorders, the Ehlers-Danlos syndrome and other connective-tissue disorders affect cardiac valves and vessels. In addition to the relatively common inherited lipoprotein disorders familial hypercholesterolaemia and familial combined hyperlipidaemia, an important dominantly inherited lipid variable contributing to coronary risk is lipoprotein(a). The gene is localized to chromosome 6 and there is full expression in childhood. Elevated lipoprotein(a) levels contribute to the occurrence and severity of early-onset coronary disease and add to the already enhanced risk in patients with familial hypercholesterolaemia. PMID- 12889665 TI - Classical homocystinuria: vascular risk and its prevention. AB - Homocystinuria due to cystathionine beta-synthase deficiency is the second most treatable aminoacidopathy. The reported incidence varies from 1 in 344,000 worldwide to 1 in 65,000 in Ireland. Untreated patients with homocystinuria have severe hyperhomocysteinaemia. Amongst its pathological sequelae, which include mental retardation, ectopia lentis and osteoporosis, vascular events remain the major cause of morbidity and mortality in untreated patients. Recognized modalities of treatment include pyridoxine, in combination with folic acid and vitamin B12; methionine-restricted, cystine-supplemented diet; and betaine. The natural history of vascular events is such that half will have an event before age 30 years and there is a predicted one event per 25 years at the time of maximal risk. In 158 patients with 2822 patient-years of treatment, there would be a predicted 112 events if left untreated, but instead only 17 vascular events were recorded during treatment (relative risk 0.09, 95% CI 0.036 to 0.228; p < 0.0001). Appropriate chronic treatment to lower hyperhomocysteinaemia is effective in reducing the potentially life-threatening vascular risk in patients with homocystinuria. These findings may also have relevance to the significance of mild hyperhomocysteinaemia that is commonly found in patients with premature vascular disease. PMID- 12889666 TI - Lipoprotein disorders and cardiovascular risk. AB - Disorders of lipoproteins often lead to disease in humans. Most often the sequelae of long-term dyslipoproteinaemia lead to atherosclerotic vascular disease in all arterial beds. Plasma elevation of low-density lipoprotein cholesterol (LDL-C), very low-density lipoproteins (VLDL) and lipoprotein(a), and reduced levels of high-density lipoproteins (HDL-C) are risk factors for coronary artery disease. Severe elevations of plasma triglycerides may lead to acute pancreatitis. In Western societies and in emerging economies, lifestyle contributes to the expression of lipoprotein disorders. Many dyslipoproteinaemias have a genetic aetiology. This review will examine the contribution of genetic lipoprotein disorders in human disease. Emphasis will be placed on monogenic disorders that are associated with coronary artery disease and novel causes of disorders of high-density lipoproteins. The consideration of screening and treatment of affected individuals, especially children, must take into account the severity of the phenotype, the long-term risk of developing vascular disease and available evidence of clinical benefit in a group of diseases that are mostly asymptomatic until manifestations of organ ischaemia in the heart, limbs or brain. PMID- 12889667 TI - Choices for treatment of hyperlipidaemia. AB - A number of studies have established that lowering of serum cholesterol levels leads to striking reductions in cardiovascular events. Although children almost never experience myocardial infarction, stroke or coronary death, paediatric autopsy studies suggest that atherosclerosis starts in childhood. Moreover, the extent of atherosclerotic change in children can be correlated with the same risk factors, including serum cholesterol level, as identified in adults. As a result of these lines of evidence, the paediatric health care community has produced guidelines for treatment of children with hypercholesterolaemia, but has also debated the pros and cons of treatment. This controversy is unlikely to be resolved completely, because definitive studies would require several decades of follow-up and would raise serious ethical concerns. The purpose of this review is to summarize current views on the optimal approach to management of hyperlipidaemia in children, given current knowledge. We suggest that the child's sex, serum cholesterol level and family history of premature cardiovascular disease may be used to stratify risk. Children at exceptional high risk may be considered for pharmacological treatment. PMID- 12889668 TI - Clinical characteristics and diagnostic clues in inborn errors of creatine metabolism. AB - Creatine deficiency syndromes are a newly described group of inborn errors of creatine synthesis (arginine:glycine amidinotransferase (AGAT) deficiency and guanidinoaceteate methyltransferase (GAMT) deficiency) and creatine transport (creatine transporter (CRTR) deficiency). The common clinical denominator of creatine deficiency syndromes is mental retardation and epilepsy, suggesting the main involvement of cerebral grey matter (grey matter disease). Patients with GAMT deficiency exhibit a more complex clinical phenotype with dystonic hyperkinetic movement disorder and epilepsy that in some cases is unresponsive to pharmacological treatment. The common biochemical denominator of creatine deficiency syndromes is cerebral creatine deficiency which is demonstrated by in vivo proton magnetic resonance spectroscopy. Measurement of guanidinoacetate in body fluids may discriminate GAMT (high concentration), AGAT (low concentration) and CRTR (normal concentration). Further biochemical characteristics include changes in creatine and creatinine concentrations in body fluids. GAMT and AGAT deficiency are treatable by oral creatine supplementation, while patients with CRTR deficiency do not respond to this type of treatment. Further recognition of patients will be of major importance for the estimation of the frequency, for the understanding of phenotypic variations and for treatment strategies. PMID- 12889669 TI - X-linked creatine transporter defect: an overview. AB - In 2001 we identified a new inborn error of metabolism caused by a defect in the X-linked creatine transporter SLC6A8 gene mapped at Xq28 (SLC6A8 deficiency, McKusick 300352). An X-linked creatine transporter defect was presumed because of (1) the absence of creatine in the brain as indicated by proton magnetic resonance spectroscopy (MRS); (2) the elevated creatine levels in urine and normal guanidinoacetate levels in plasma, ruling out a creatine biosynthesis defect; (3) the absence of an improvement on creatine supplementation; and (4) the fact that the pedigree suggested an X-linked disease. Our hypothesis was proved by the presence of a hemizygous nonsense mutation in the male index patient and by the impaired creatine uptake by cultured fibroblasts. Currently, at least 7 unrelated families (13 male patients and 13 carriers) with a SLC6A8 deficiency have been identified. Four families come from one metropolitan area. This suggests that SLC6A8 deficiency may have a relatively high incidence. The hallmarks of the disorder are X-linked mental retardation, expressive speech and language delay, epilepsy, developmental delay and autistic behaviour. In approximately 50% of the female carriers, learning disabilities of varying degrees have been noted. PMID- 12889670 TI - Duct tape, please. PMID- 12889671 TI - The anatomical relationship between the position of the auriculotemporal nerve and mandibular condyle. AB - Head, neck, face, and ear pains are commonly associated with disorders of the temporomandibular joint (TMJ). Several theories have been proposed regarding the functional relationship of the TMJ and the associated structures, and how they might contribute to certain painful conditions. This study was conducted to determine the anatomic relationship of the auriculotemporal nerve to the middle meningeal artery and the mandibular condyle. Forty human cadaver temporomandibular joints were dissected to locate the precise position of the auriculotemporal nerve to the mandibular condyle. The study findings revealed a significant variation in the relationship of the auriculotemporal nerve to the middle meningeal artery. The auriculotemporal nerve was found to be between 10-13 mm inferior to the superior surface of the condyle and 1-2 mm posterior to the neck of the condyle. The nerve was not found to be in a position that would likely create an entrapment with adjacent tissues. These findings may assist the clinician to locate the most appropriate injection site for an auriculotemporal nerve block. PMID- 12889672 TI - Maxillomandibular relationship in TMD patients before and after short-term flat plane bite plate therapy. AB - This study assessed the maxillomandibular relationship in temporomandibular disorders (TMD) patients, before and after short-term, flat plane bite plate therapy. It was of interest to determine the incidence and degree of mandibular deviation in a group of TMD patients and whether the mandible would shift to the midline and consequently affect reported symptoms. Seventeen female and three male subjects (age range 19-60) were included in the study. Thirteen subjects were diagnosed with myofascial pain while seven were diagnosed as exhibiting disk displacement with reduction (Research Diagnostic Criteria). After taking impressions for these subjects, casts were fabricated and mounted. Maxillomandibular relationship was evaluated by the Denar Centric Check system (Anaheim, CA). The maxillary and mandibular labial frena were used as a reference to evaluate mandibular shift. Symptom questionnaires were used to assess temporomandibular joint pain and clicking. All subjects exhibited deviation (12 subjects to the right and 8 subjects to the left) prior to bite plate therapy. After flat plane bite plate therapy, the mandibular position of all subjects shifted toward the labial frenum midline position. Based on the Binomial test, the shift was significant (p < 0.001). Measurements on the Centric Check system showed a significant movement of both condyles in the anterio-posterior plane as well as the vertical plane. There was also significant reduction in TMJ pain and clicking (p < 0.01). The results support the hypothesis that the balanced position of the mandible is with frena aligned. When occlusal obstructions are eliminated, the mandible will drift to this position. PMID- 12889673 TI - The prevalence of trigeminal neuritis with TMD. AB - Subjects with temporomandibular joint disorder (TMD) occasionally present with additional orofacial pain complaints. These can arise from dysfunction in teeth, bones, ligaments, tendons, nerves, and other structures. In this retrospective study, a group of 501 consecutive subjects with TMD complaints were evaluated for the presence of trigeminal neuritis. Very little information on the prevalence of this condition concomitant with TMDs exists in the literature. The existence of trigeminal neuritis was determined by the presence of pain when palpating trigeminal peripheral nerve branches exiting the supraorbital, infraorbital, and mental foramina in addition to the supratrochlear nerve. Each subject in this study had the involved nerves blocked with local anesthesia injections or lidocaine iontophoresis to assist in confirming the source of pain. Sixty subjects with TMD were found to have trigeminal neuritis. Early recognition of this disorder is important because treatment is usually more successful in the acute peripheral state. PMID- 12889674 TI - Measurement of muscle hardness using a hardness meter: application to the masseter and temporal muscles and reproducibility of measurement. AB - This study was designed to evaluate the intra-examiner and inter-examiner reliability of measurements of masticatory muscle hardness and to confirm that the muscle hardness increases with contraction using a commercially available muscle meter. Twenty healthy asymptomatic female subjects participated in this study. Hardness was expressed as numerical relative values (0-100). First, muscle hardness was measured at a standardized point located in the masseter muscle and temporal muscle in a randomized order by two examiners, and again by one of the same examiners after ten minutes for the reproducibility study. Then the muscle hardness was measured at each point for 0 kgf, 3 kgf, 6 kgf, and 9 kgf levels of bite force. As a result, intraclass correlation coefficient (ICC) analysis revealed good intra-examiner reliability in the masseter muscle (ICC = 0.711), good intra-examiner reliability in the temporal muscle (ICC = 0.643), good inter examiner reliability in the masseter muscle (ICC = 0.631), and unacceptable inter examiner reliability in the temporal muscle (ICC = 0.008). Also, our results showed that muscle hardness increased with contraction, and relationships with a slope of 1.229, a y-intercept of 62.513, and a correlation coefficient of 0.448 were observed in the masseter muscle. However, no correlation was found between muscle hardness and bite force in the temporal muscle. The findings indicate that measurement of hardness provides reliable physiological information about the masseter muscle in this setting. PMID- 12889675 TI - Convergence defects in patients with temporomandibular disorders. AB - The aim of this study is to show the presence of a correlation between ocular convergence defects (OCD) and temporomandibular disorders (TMD) among a group of adult subjects. The group studied was made up of 48 subjects (12 males and 36 females). The average age was 35 with a range of 19-45 years of age. The subjects presented with TMD and muscular pain and/or dysfunction. Forty-eight subjects with TMD for the case study were matched by gender and age to 48 control subjects seeking routine dental care (control group). All the subjects were examined by the same orthoptist who classified the ocular convergence degree using two tests. The first test evaluated the distances (in centimeters) of the convergence near point (3-4 cm: normal; 5-7 cm: sufficient; > 7 cm: insufficient). The second test assessed the fusional convergence using a Berens prism test (> 25 diopters: normal; between 18-25: sufficient; < 18 diopters: insufficient). In the TMD group, 36 subjects (75%) showed a compromise of convergence: 13 (36%) were classified in the 5-7 degree range and 23 (48%) in the > 7 cm degree range. The Berens test showed ten subjects (28%) in the group < 18D and 26 (72%) in the group 18-25D. The control-group presented ten (21%) subjects with compromise of convergence: three classified in the group < 18D and seven in the group 18-25D. The TMD subjects presented a higher statistical percentage (p < 0.0001) of ocular convergence defects. The TMD patients also reported a strong association referred to specific signs and symptoms, i.e., limited maximal opening or myofascial pain. There were some subjective reports also of headaches and torcicollis (neck stiffness) which appeared significantly more frequently in subjects with a compromise of convergence. The study showed a much higher prevalence of ocular convergence defects in patients with head, neck, and shoulder pain. PMID- 12889676 TI - Normal and pathological anatomy of the TMJ viewed by computerized panoramic arthroscopic images. AB - The individual single images obtained to document TMJ arthroscopy are often difficult to interpret. The aim of this paper is to make available a new computer aided image process system to reformat the panoramic arthroscopic images of the temporomandibular joint (TMJ). Arthroscopic images were input directly into a computer and memorized into a magneto optical disk. Using Photoshop 5.0 (Adobe Systems, Inc., Mountain View, CA) software for Windows 98 (Microsoft Corp., King County, WA) the images were edited and adjusted to compound sagittal and/or coronal panoramic images of normal articular surfaces as well as pathology of TMJ disorders. Normal TMJ sagittal and coronal two-dimensional composite panoramic arthroscopic images were achieved. The panoramic images of some intracapsular disorders (including disk displacement, osteoarthrosis, adhesion, and disk perforation) were also obtained. The computerized arthroscopic panoramic images are a new technique that may aid in the understanding of TMJ anatomy and TMJ disorders. PMID- 12889677 TI - The functional relationship between the craniomandibular system, cervical spine, and the sacroiliac joint: a preliminary investigation. AB - The hypothesis of a functional coupling between the muscles of the craniomandibular system and the muscles of other body areas is still controversial. The purpose of this pilot study was to examine whether there is a relationship between the craniomandibular system, the craniocervical system and the sacropelvic region. To test this hypothesis, the prevalence and localization of dysfunction of the cervical spine and the sacroiliac joint were examined in a prospective, experimental trial. Twenty healthy students underwent an artificial occlusal interference, which caused an occlusal interference. The upper cervical spine (CO-C3) and the sacroiliac joint were examined before, during and after this experimental test. The primary outcome with these experimental conditions was the occurrence of hypomobile functional abnormalities. In the presence of occlusal interference, functional abnormalities were detected in both regions examined and these changes were statistically significant. The clinical implications of these findings may be that a complementary examination of these areas in CMD patients could be useful. PMID- 12889678 TI - Orthosis thickness and its effect on temporomandibular joint space. AB - The object of this study was to determine the effect of varying thicknesses of orthoses on temporomandibular (TM) joint space. The TM joint space was measured using the centric cut view of tomographic images. The measurements used selected fixed landmarks of the joint complex anatomy to find joint space. The fixed points used for the measurement of the joint space are no different than the fixed points methodology used in orthodontic cephalometric measurements. The placement of the fixed points on a radiograph is subjective. Degrees of accuracy can be accomplished with consistent use just as in the practice of cephalometric measurements. Results of such measurements do provide a relatively true record of the actual condyle and fossa relationship. The study concluded that the thickness of the orthosis moved the condyle downward and horizontally without regard to its original positioning. If clinicians know the condylar position before and after orthosis placement, a concentric position of the condyle can be established to assist in the patient's second phase treatment. Further research will prove that the physiological and anatomical balance of the temporomandibular joint complex is important in effectively managing craniofacial pain patients. PMID- 12889679 TI - Clinical evaluation of amitriptyline for the control of chronic pain caused by temporomandibular joint disorders. AB - Temporomandibular disorder (TMD) is characterized by a combination of symptoms affecting the temporomandibular joint and/or chewing muscles. The two most common clinical TMD symptoms are pain and dysfunction. Pain is usually caused by dysfunction, and emergency therapy has focused on controlling it. Recent investigations into TMD have led to the recommendation of antidepressants as a supporting treatment against constant neuralgic pain. The aim of this double blind study was to verify the efficiency of antidepressants (amitriptyline) as a support in the treatment of chronic TMD pain. Twelve female volunteers presenting chronic TMD pain were divided into two groups and treated for 14 days: Group 1 with 25 mg/day of amitriptyline and Group 2 with a placebo. The intensity of pain and discomfort was evaluated daily, using a visual analog scale (VAS), over a period of seven days preceding the treatment (baseline), during the 14-day treatment, and for seven days after the treatment. The results revealed a significant reduction of pain and discomfort in Group 1 (75%) compared to Group 2 (28%) during the three weeks beginning at baseline (p < 0.01). Amitriptyline proved to be an efficient alternative treatment for chronic pain in TMD patients. PMID- 12889680 TI - Traumatic arthritis associated with acute dislocation of the TMJ: a case report. AB - An unusual case of traumatic arthritis associated with acute dislocation of the temporomandibular joint (TMJ), successfully treated by arthroscopy, is reported. The intraarticular pathology is described and the management of arthroscopic lysis and lavage for traumatic arthritis of the TMJ is discussed. PMID- 12889681 TI - Explanations for the smoking paradox in Japan. AB - The prevalence of cigarette smoking among Japanese men has been consistently high compared with Western males over the past 30 years. However, during the same period, the incidence of and mortality rates for lung cancer have consistently been lower in Japan than in Western countries ('Japanese smoking paradox'). The odds ratio/relative risk of cigarette smoking for lung cancer mortality/incidence relative to the same number of cigarettes smoked per capita in Japan, were apparently lower than those in Western countries. This must be the cause driving the 'Japanese smoking paradox'. Furthermore, low carcinogenic ingredients in Japanese cigarettes and a congenitally-related resistance to smoking-related lung carcinogenesis emerged as the main factors which have brought the 'Japanese smoking paradox'. PMID- 12889682 TI - Determination of ethnicity in children in The Netherlands: two methods compared. AB - The aim of this study was to investigate whether names (surnames and the combination of surnames and first names) are adequate to identify Turkish, Moroccan and Surinamese children in a Dutch population. Parents visiting the Emergency Department of the Sophia Children's Hospital with their child filled in a questionnaire about their ethnic background, socio-economic factors and cultural factors. Patients' names were classified by students with a Turkish, Moroccan and Surinamese (Hindustanic and Creole) background. A combined name method (surnames in combination with first names) and surnames only were compared with a reference method (ethnicity by parents' country of birth). The validity of both name methods and the measurements of agreement (kappa) of both comparisons were analysed. Turkish names showed good validity measurements between the combined name-method and the reference method (sensitivity of 81% and a positive predictive value of 86%). The Arabic names were distinguishable with a sensitivity of 77%, specificity of 99% and positive predictive value of 87%. Surinamese names were less easy to distinguish from the other names (69, 96 and 69% respectively). Overall, the measurement of agreement showed a kappa of 0.69 in the comparison between the classification of the combined names and the classification by parent's country of birth. The classification of the combined names was slightly better than the classification by surnames only. CONCLUSIONS: Names can be used to identify Turkish and Arabic (Moroccan) children in a routinely registered database of Dutch children. For Surinamese names this method is less suitable. PMID- 12889683 TI - The sense of coherence, occupation and all-cause mortality in the Helsinki Heart Study. AB - BACKGROUND: The aim of this study was to test the hypothesis that people with a strong sense of coherence (SOC) have decreased all-cause mortality. METHODS: The effect of occupation and the SOC on all-cause mortality was studied among 4405 Finnish middle-aged employed men in a prospective 8-year follow-up study. RESULTS: Using Cox proportional hazards models the crude relative risk for all cause mortality for the low SOC tertile when compared to the high SOC tertile was 1.23 (95% CI: 0.90-1.68). Adjusting for age, smoking, alcohol and occupation increased the risk slightly to 1.35. Occupation was an effect modifier, since among white-collar workers the corresponding relative risk of the low SOC tertile was 2.27 (95% CI: 1.12-4.59, p = 0.02) and among blue-collar workers the relative risk for all-cause mortality was stable (1.33-1.52) in each SOC tertile. The classic risk factors, smoking and alcohol, showed higher relative risks than the SOC. CONCLUSIONS: The effect of the health-promoting qualities of the SOC upon all-cause mortality was significant among white-collar workers, but not among blue-collar workers. PMID- 12889684 TI - The size of educational differences in mortality from specific causes of death in men and women. AB - This study examines the association between education and mortality from specific causes of death based on mortality records for 1996 and 1997, and 1996 population census data from the Region of Madrid (Spain). Poisson regression models were used to estimate the percentage increase in mortality associated with 1 year less education. The percentage increases in mortality from stomach cancer, lung, bladder and liver cancers, for aids, chronic obstructive pulmonary disease, pneumonia and influenza, and chronic liver disease and cirrhosis were higher in men than in women, whereas the percentage increases in mortality from colon cancer, diabetes mellitus, ischemic heart disease and nephritis, nephrosis and nephrotic syndrome were higher in women. The results found for some causes of death--lung cancer, ischemic heart disease, diabetes mellitus and chronic obstructive pulmonary disease--reflect the variations by educational level in the prevalence of lifestyle-related risk factors in men and women. Various hypotheses have been suggested for other causes of death, but it is not known why the magnitude of the association between education and mortality from some causes of death differs between men and women. Future studies of this subject may provide some clues as to the underlying mechanisms of this association. PMID- 12889685 TI - Relation of coffee, green tea, and caffeine intake to gallstone disease in middle aged Japanese men. AB - A possible protective effect of coffee or caffeine intake in the formation of gallstones has been suggested in some epidemiological studies. We examined the relation of coffee, green tea, and caffeine intake to gallstone disease in middle aged Japanese men, distinguishing known gallstones from unknown diagnosed gallstones. Study subjects were 174 cases of gallstones as determined by ultrasonography, 104 cases of postcholecystectomy, and 6889 controls of normal gallbladder in the total of 7637 men who received a health examination at four hospitals of the Self-Defense Forces (SDF). Of the 174 cases of prevalent gallstones, 50 had been aware of having gallstones. Previously diagnosed gallstones and postcholecystectomy were combined as known gallstone disease. The consumption of coffee and green tea was ascertained by a self-administered questionnaire, and caffeine intake was estimated. Statistical adjustment was done for body mass index, smoking, alcohol use, rank in the SDF, and hospital. Coffee and caffeine intake were associated each with a statistically significant increase in the prevalence odds of known gallstone disease, but unrelated to newly diagnosed gallstones. Adjusted odds ratios of known gallstone disease were 1.7 (95% confidence interval [CI] 1.1-2.8) for coffee consumption of five cups or more per day vs. no consumption and 2.2 (95% CI: 1.3-3.7) for caffeine intake of 300 mg/day or more vs. less than 100 mg/day. The consumption of green-tea showed no material association with either unknown gallstones or known gallstone disease. The findings do not support a hypothesis that coffee drinking may be protective against gallstone formation. PMID- 12889686 TI - Lipid-lowering therapy and cholesterol levels following acute myocardial infarction: a German study of 5361 patients. AB - BACKGROUND: Current studies on secondary prevention of cardiovascular events (CARE, LIPID, 4S) illustrate the necessity of an effective lipid-lowering therapy. An important part of secondary prevention is the prompt measurement of lipids following an infarct, to be able to start therapy as quickly as possible. AIMS: This study should show the general situation in the prescription of lipid lowering drugs in patients with acute myocardial infarction (MI) and if there are gender differences in therapy and in the lipid parameters. The second aim was to determine the therapeutical conclusions for secondary prevention made in-hospital in patients with heart attacks. METHODS: Post-infarct lipid parameters (total cholesterol (TC), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C), triglycerides (TG) were examined in 5361 patients within 24 hours following an acute MI. Lipid parameters from 576 patients were measured again after 1 week. RESULTS: 80.1% of men under examination and 81.9% of the women had suffered from their first MI. Only 9% of all patients were already under lipid-lowering therapy before the MI. After the MI TC and LDL-C levels decreased significantly in a time-dependent manner. Around 50% of patients received lipid-lowering drug, or the recommendation of one, during their hospital stay. CONCLUSION: The range of TC values of the patients examined was comparable to those in the CARE and the 4S secondary prevention studies. It can therefore be assumed that the results of these studies are also applicable to Germany. Nevertheless, according to existing data, therapy with lipid-lowering drugs is currently unsatisfactory, even in secondary prevention. PMID- 12889687 TI - Comparing the results of a cholesterol and saturated fat screener when using two different scores. AB - Screener are useful instruments for categorising individuals according to their nutrient intake. To accurately classify the individuals it is important to correctly analyse the screener. Therefore we compared a simple and a weighed score of a cholesterol and saturated fat (SF) screener with the cholesterol and SF intake, respectively, calculated from an extensive food frequency questionnaire. The validation results did not change when a weighed score was used instead of the simple one, indicating that a simple score does not introduce a higher amount of misclassification. PMID- 12889688 TI - Reproducibility over time of measurements of androgens, estrogens and hydroxy estrogens in urine samples from post-menopausal women. AB - Sex steroid concentrations in urine samples from post-menopausal women have been associated with risk of various chronic diseases. The basic requirement for the assessment of risk in such large-scale epidemiological studies is that subjects be ranked accurately by their average, long-term hormone levels. We examined the reproducibility over time of measurements of urinary testosterone (T), 5alpha androstane-3alpha, 17beta-diol (ADIOL), estrone (E1), estradiol (E2), 2-hydroxy estrone and 2-hydroxy estradiol, (2(OH)-E), 16alpha-hydroxyestrone (16alpha(OH) E1) and the ratio of 2(OH)-E and 16alpha(OH)-E1, in a representative sub-sample of post-menopausal women (n = 43) participating in an ongoing prospective cohort study. Women collected three first morning urine voids on different occasions, with average time difference between the first and the third urine sample of 5.1 years. T, ADIOL, E1 and E2 were measured by radio immunoassay after enzymatic hydrolysis, solid-phase extraction and HPLC purification of the samples, while 2(OH)-E and 16alpha(OH)-E1 were assayed by solid-phase enzyme immunoassay after enzymatic hydrolysis. Intra-class correlation co-efficients (ICCs) over time were very good for T (r = 0.85), acceptable for E2, E1 and ADIOL (r > 0.55), but low for 2(OH)-E, 16alpha(OH)-E1 and their ratio (r < 0.46). The adjustment for creatinine concentrations did not increase these correlations. PMID- 12889689 TI - Infections and risk of type I diabetes in childhood: a population-based case control study. AB - OBJECTIVE: This study focuses on the evaluation of some infectious diseases as risk determinants of type I diabetes mellitus (DM). METHODS: A population-based case-control study was carried out by referring to the type I DM population-based register of the Abruzzo region of Italy as it includes all type I DM cases since January 1 1990, the point at which the register became operative. The pediatric population (age: 0-14), living in the same municipalities of the cases, was selected as the control population. Data were collected through questionnaires submitted by a physician to parents of cases and controls. Conditional logistic regression models were used to evaluate association between determinants and onset of type I DM. RESULTS: The risk of diabetes for children exposed to only one infection (morbilli, parotitis, rubella, pertussis or varicella) is not statistically significant: OR: 0.778; CI: 0.427-1.370. On the contrary, when two infections are contracted statistically significant results occur: OR: 2.375; CI: 1.149-4.914; for more than two infections values are: OR: 6.786; CI: 2.881 17.877. No substantial difference in odds ratios (ORs) after adjustment for confounding variables was found. A significant decrease in OR was noted for pertussis and MMR vaccinations, respectively: OR: 0.015; CI: 0.001-0.251; OR: 0.400; CI: 0.201-0.799. CONCLUSIONS: Since the higher the number of contracted infections, the higher the risk of diabetes, contracted infections can be considered potential accelerating factors of clinical manifestation of type I DM. Therefore multiple exposures might speed up the onset of diabetes in children. This study suggests the utility of applying the risk model method to wider populations, especially if the geographical variability of standardised incidence rates of type I DM in pediatric age is taken into consideration. PMID- 12889690 TI - Can respiratory syncytial virus etiology be diagnosed clinically? A hospital based case-control study in children under two years of age. AB - An unmatched, hospital-based case-control study was performed, to determine, whether respiratory syncytial virus (RSV) etiology in hospitalized young children can be predicted clinically. Children under 2 years of age admitted with a lower respiratory tract infection in three hospitals in northern Germany were included (one tertiary and two secondary centers). Cases were children tested positive for RSV by multiplex RT-PCR. One control group consisted of children tested negative for RSV in the multiplex-RT-PCR and a second control group consisted of patients in whom no PCR was done. A weighted backward stepwise logistic regression model was applied for multivariate analysis. RSV-etiology could be predicted with a sensitivity of 72.8% and a specificity of 73.2%. Young age, disease entity- pneumonia or bronchiolitis, center, intercostal retractions, absence of an underlying condition, low level of C-reactive protein, short duration of symptoms (all on admission), prematurity and epidemiologic year were predictive; anatomical infiltrates and wheezing were not. Pathogen specific diagnosis is necessary for individual therapy, allocation in observational studies or treatment trials and for surveillance of airway infections in children, since the positive predictive value is too low for an accurate diagnosis and decision making. Multivariate techniques are effective tools in complex clinical research for deconfounding. PMID- 12889692 TI - Prevalence of antibodies to spotted fever group rickettsiae in the region of Split (southern Croatia). AB - The prevalence of IgG antibodies to spotted fever group rickettsiae (SFG) was investigated by the indirect immunofluorescence assay in a sample of 1207 healthy residents of the central part of southern Croatia (eastern coast of Adriatic Sea). Sera obtained from subjects living in three different places of residency (urban, suburban and rural environs of the region) were tested. Overall 528 (43.7%) persons had significant antibody titers (1:40 or higher), with a significantly higher prevalence in suburban and rural areas. The results show that inhabitants in the area are clearly being exposed to SFG rickettsiae. Risk factors significantly associated with seropositivity to SFG rickettsiae were: living in suburban and rural settlements (odds ratio [OR] = 1.81; 95% confidence interval [CI] = 1.35-2.4), contact with animals (OR: 1.52; 95% CI: 1.15-1.99), and increase of age (OR: 1.18; 95% CI: 1.09-1.26). PMID- 12889691 TI - Tattooing and transfusion-transmitted diseases in Brazil: a hospital-based cross sectional matched study. AB - BACKGROUND: Presence of tattoos has been a criterion for temporary deferral of blood donors. Scientific evidence remains equivocal regarding the association between tattooing and transfusion-transmitted diseases (TTDs). METHODS: A cross sectional matched study was undertaken among adults attending a Brazilian hospital and blood bank. The exposure of interest was having at least one permanent tattoo, and the outcomes were the presence of serological markers for the following TTDs: hepatitis B virus (HBV), hepatitis C virus (HCV) and human immunodeficiency virus (HIV) infections, syphilis, and Chagas' disease. Exposed and unexposed subjects were matched on age, sex, and main clinical complaint. Associations were assessed by odds ratios (ORs), adjusted for confounders by unconditional logistic regression. FINDINGS: The study recruited 345 subjects, 182 with tattoos. Having a tattoo was associated with HCV (OR: 6.41; 95% confidence interval (CI) 1.29, 31.84), and with having at least one positive test for any TTD (OR: 2.05, 95% CI: 1.11, 3.81). No statistically significant associations were found between tattooing and HBV or HIV infection, syphilis or Chagas' disease, but these results are inconclusive given the large CI obtained. INTERPRETATION: Having a tattoo is not an important indicator for testing positive for a TTD, except for HCV infection. Taking into consideration the increasing prevalence of tattooing in the general population, the absolute need of a safe and sustainable blood supply and optimization of the cost-effectiveness of screening blood donors, further research on tattoos is urgently required. PMID- 12889693 TI - Evidence of Toscana virus circulation in Umbria: first report. AB - To evaluate, for the first time, circulation and clinical expression of Toscana virus (ToSv) in Umbria region we studied: (1) 93 cases of aseptic meningitis and meningoencephalitis admitted to our Department from 1989 to 2001 with negative results for common neurotropic virus; (2) 50 healthy subjects. Specific antibodies (IgM and IgG) anti-TOSv were found in 36.6% of aseptic meningitis, in 6.06% of meningoencephalitis and (IgG) in 16% of healthy subjects. PMID- 12889694 TI - Inaccuracies in relating 25-hydroxyvitamin D to ischemic heart disease. PMID- 12889695 TI - Primary incisor restoration. PMID- 12889696 TI - The effects of chlorhexidine gel on Streptococcus mutans infection in 10-month old infants: a longitudinal, placebo-controlled, double-blind trial. AB - PURPOSE: This study investigated the long-term effects of 0.2% chlorhexidine gel, used as a weekly brush-on gel, on Streptoccocus mutans infection in 10-month-old infants. METHODS: The investigation followed the criteria of a placebo controlled, double-blind, longitudinal clinical trial. Infants were recruited at birth and oral microbiological swabs were taken at 3 monthly intervals, together with medical, dental, dietary and brushing histories. Children who were found to be colonized with S. mutans were randomly assigned to either the chlorhexidine gel group (N = 50) or placebo gel group (N = 46), and parents were instructed to brush the gel on the teeth once per week for 12 weeks. In another control group (N = 210), infants did not use either chlorhexidine or placebo gels. Saliva samples were cultured using S. mutans-selective tryptone-yeast extract-cysteine sucrose-bacitracin (TYCSB) agar. The mean age of the children was 10.2 +/- 2.6 months at the start of the trial and subjects were followed until the ages of 18 months. RESULTS: In the children with initial low S. mutans counts of < 300 CFU/mL, there was a significant percentage reduction in S. mutans counts in the chlorhexidine-gel group compared to the placebo gel and no-gel control groups after 3 months of weekly gel brushing. However, no significant differences with the placebo group were observed after 15 months of follow-up. There were 39 children (41%) who achieved reduction of their S. mutans to 0 CFU/mL. Compared to those who remained infected with S. mutans, these children had higher toothbrushing frequencies (P < .001) and toothpaste use (P < .001), as well as lower frequencies of daily feeds (P < .01), and lesser weekly frequencies of sweet solids and liquids (P < .001). CONCLUSIONS: Children with relatively low initial S. mutans counts (< 300 CFU/mL) showed a reduction in S. mutans counts in the first 3 months when 0.2% chlorhexidine gel was brushed on the teeth weekly. No differences were observed when compared with the placebo and no-treatment groups at later follow-up periods. PMID- 12889697 TI - Effects of oral health education and tooth-brushing on mutans streptococci infection in young children. AB - PURPOSE: The purpose of this study was to examine the effects of maternal dental health education and tooth-brushing instruction on the levels of mutans streptococci in pre-school children. METHODS: One hundred and seven children (44 boys and 63 girls) with a mean age of 20.5 months, randomly selected from a community child health clinic within a 2-week period, participated in this study. Medical, dental, and dietary information was obtained through a questionnaire. The children's mouths were examined, and plaque samples were obtained by swabbing the teeth and mucosa. A commercial microbiological kit was used to determine the presence of mutans streptococci. The mothers were instructed in tooth-brushing using a soft-scrub method. The children were recalled to the same clinic after a period of 4 weeks to obtain a second evaluation of the mutans streptococci levels. RESULTS: At the first visit, 69 of the 107 children (64%) showed positive infection with mutans streptococci. Ninety (84%) children returned for the recall examination. In the second examination, only 44 (49%) of the 90 children showed positive results for mutans streptococci. The difference in number of children who tested positive is statistically significant (P < .01). At the first visit, children who did not show infection with mutans streptococci were those who reported greater frequency of tooth-brushing (P < .05) and less snacking (P < .05). There were 26 children (29%) who converted from positive to negative results for mutans streptococci infection between the first and second visits (P < .01). This conversion from positive to negative infection was attributed mainly to increased tooth-brushing, as other dental health habits remained the same. CONCLUSIONS: Infection with mutans streptococci in young children is associated with increased snacking frequency and inadequate tooth-brushing. A single dental health education session and tooth-brushing instruction to mothers results in approximately a 25% reduction in mutans streptococci infection in young children from a relatively high socioeconomic status. PMID- 12889698 TI - Progression of proximal caries in the mixed dentition: a 4-year prospective study. AB - PURPOSE: The purpose of this study was to investigate the survival rate and median survival time of different stages of proximal caries in 196 children aged 6 to 8 years at baseline examination. METHODS: Proximal caries and its progression were diagnosed from bitewing radiographs taken at 1-year intervals over a period of 4 years. The mesial surface of the first permanent molars, the mesial and distal surfaces of the first and second primary molars, and the distal surface of the primary canine were examined. Sound surfaces, caries lesions in the external- and internal-half of the enamel and external-, middle-, and internal-third of the dentin, as well as filled, extracted, and exfoliated teeth were recorded. A life table analysis was performed to estimate the annual and cumulative survival rates and the median survival time of each state of proximal lesions. RESULTS: For the sound mesial surfaces of the first permanent molars, the cumulative survival rate was 76% and the median survival time was over 48 months. The corresponding values for the external-half of the enamel lesions were 41% and 45 months. Regarding the sound mesial and distal surfaces of the primary teeth, the cumulative survival rate was 92% and the median survival time was over 48 months. The corresponding values for the external and internal-half of the enamel lesions were 40% and 31 months, and 29% and 22 months, respectively. The value of the cumulative survival rate for the external-third of dentin lesions was 42%, with a median survival time of 34 months, and the value for the middle third of the dentin was 38% with a median survival time of 17 months. CONCLUSIONS: The risk of developing carious lesions in the sound proximal surfaces of both primary teeth and first permanent molars is low during the mixed dentition period; the progression of the external-half of enamel lesions of the first permanent molars is low for the first 3 years and becomes faster thereafter; and as the time period that the tooth exposed to cariogenic factors increases, the progression of the proximal caries is faster for all stages of the lesion in the primary teeth. PMID- 12889699 TI - Prevalence of dental caries and enamel defects in Connecticut Head Start children. AB - PURPOSE: The aim of this study was to determine the prevalence of dental caries and enamel defects in a sample of predominantly African American and Hispanic children from an inner city Head Start program located in Connecticut. METHODS: A total of 517 children were examined for dental caries and enamel defects. Children's caries experience was described using the dmfs/t indices, and dental defects were described using a modified developmental defects of enamel index. RESULTS: The mean dmfs was 3.0, and 38% of the children had caries. The prevalence of enamel defects was 49%. When analyzed by race/ethnicity no significant differences in dmfs scores or the prevalence of defects were observed. The majority of defects were located on anterior teeth, and the type of defect varied with the location. On the buccal surface of canines, hypoplasia accounted for 70% of the lesions. On maxillary anterior teeth, linear opacities accounted for 50% of the lesions. A positive association between enamel defects and caries was observed. CONCLUSIONS: The prevalence of caries and defects in these Head Start children was high, with most defects located on anterior teeth. Enamel defects were associated with an increased caries incidence. PMID- 12889700 TI - Treatment of crown fractures with pulp exposure in primary incisors. AB - The treatment options of enamel dentin crown fractures with pulpal exposure in the primary dentition traditionally consist of direct pulp capping, pulpotomy, pulpectomy, or extraction. Many clinicians disagree on the most appropriate treatment, and individual preferences exist within dentistry. Partial pulpotomy, also known as the Cvek pulpotomy, is a mode of treatment which is widely used in the permanent dentition but less so in primary teeth. This conservative technique is described and its advantages over the others are presented. In addition, a report of a case with a 2-year follow-up is also included. The purposes of this article are: (1) to present the indications and contraindications of the various treatment modalities for primary incisors with complicated crown fractures; and (2) to suggest partial pulpotomy as a conservative and more appropriate approach for primary incisors with complicated crown fracture. PMID- 12889701 TI - Marginal adaptation of stainless steel crowns. AB - The chief goal of full coronal restoration using preformed stainless steel crowns (SSC) is replication of normal crown form and function. Marginal adaptation of SSCs involves appropriate crown size selection, trimming the crown form to achieve proper length, crimping crown edges to proximate the prepared tooth, and finishing and polishing the crown form. This report about SSC restoration focuses on the procedure of adapting, finishing, and polishing crown margins. PMID- 12889702 TI - Use of nasoalveolar molding appliance to direct growth in newborn patient with complete unilateral cleft lip and palate. AB - The alignment of the alveolar segments creates the foundation upon which excellent results of lip and primary nasal surgery are dependent in the repair of the cleft lip and palate patient. The purpose of this article is to illustrate the step-by-step fabrication process of the nasoalveolar molding appliance used to direct growth of the alveolar ridge, lips, and nose in the presurgical treatment of cleft lip and palate. As a result of this appliance, the primary surgical repair of the nose and lip heals under minimal tension, thereby reducing scar formation and improving the esthetic result. PMID- 12889703 TI - Median ages of eruption of the primary teeth in white and Hispanic children from Arizona. AB - PURPOSE: The purpose of this project was to establish median eruption ages of primary teeth and evaluate eruption differences between contralateral teeth by gender, ethnicity, and household income. Data was derived from a caries study of preschool children. METHODS: A total of 4,277 white (non-Hispanic) and Hispanic preschool children from Arizona were visually examined for caries and tooth eruption status by 5 examiners between February 1994 and September 1995. Analyses of eruption status were conducted using logistic regression to produce an eruption probability distribution from which median eruption ages for the primary teeth were determined. Possible differences in eruption timing between contralateral teeth by gender, ethnicity, and household income were examined. RESULTS: Eruption status of 1 tooth was significantly predictive of the pair matched tooth for all pairs. Differences were found for the maxillary central incisor (gender) and the mandibular second molar (ethnicity). Eruption ages did not vary by household income. Median eruption ages for each of the 10 primary teeth are presented. CONCLUSIONS: This study provides contemporary median eruption ages for primary teeth. PMID- 12889704 TI - Shear bond strengths of orthodontic brackets cemented to bovine enamel with composite and resin-modified glass ionomer cements. AB - PURPOSE: The purpose of this in vitro study was to determine the effects of short and long-term storage on the shear bond strength of metal, polycarbonate, and ceramic orthodontic bracket bases using autopolymerizing resin composite and resin-modified glass ionomer cements (RMGIC). The glass ionomer cement was applied in both a wet and a dry environment. METHODS: With a method developed in the authors' laboratory, orthodontic brackets were cemented under constant pressure to embedded bovine incisor enamel. All cements were mixed and applied in accordance with the manufacturer's instructions. The specimens were stored in water at 37 degrees C for 24 hours, 7 days, or 180 days. After the lapse of each time interval, they were shear tested to failure. The shear bond strengths (SBSs) were converted to megapascals (MPa). An adhesive remnant index (ARI) was used to record the site of the residual cement. RESULTS: There were no precipitous increases or decreases in SBS over a lapsed time of 180 days, although some variations occurred between 24 hours and 7 days. Similar findings were recorded for ARI. CONCLUSIONS: The bracket base-cement combinations produce clinically sustainable SBSs over time. Selection of the cement may be important in patients who exhibit a high risk for caries. PMID- 12889705 TI - Simultaneous activation technique: an alternative for bonding composite resin to glass ionomer. AB - PURPOSE: The purpose of this study was to evaluate a new simultaneous activation technique (SAT)-for the union between glass ionomer cements and composite resins. METHODS: Forty primary molars were embedded in self-curing acrylic resin. Class I cavity preparations were performed on the dentin surface and the teeth were randomly divided into 4 groups (N = 10). Two groups were restored with the SAT and 2 by etching the ionomer surface/sandwich technique (ST). The materials tested were Vidrion F (SS/ White), Vitremer (3M), and Z100 (3M). After 24 hours in aqueous solution at 37 degrees C, a tensile test was performed. The microleakage study was also performed by selecting 40 primary molars which were distributed in the same way as in the bond strength test. Furthermore, 1 tooth from each group of the microleakage test was evaluated by means of scanning electron microscopy. RESULTS: The SAT and ST did not show statistically significant differences in both tests. CONCLUSIONS: The SAT is a less complex, quicker, and feasible alternative for the bonding of glass ionomer cements to composite resins in primary molars. PMID- 12889706 TI - Retention of veneered stainless steel crowns on replicated typodont primary incisors: an in vitro study. AB - PURPOSE: The purpose of this in vitro study was to determine the effect of crimping and cementation on retention of veneered stainless steel crowns. METHODS: One hundred twenty crowns, 90 from 3 commercially available brands of veneered stainless steel crowns (Dura Crown, Kinder Krown, and NuSmile Primary Crown) and 30 (plain) Unitek stainless steel crowns were assessed for retention. An orthodontic wire was soldered perpendicular to the incisal edge of the crowns; the crowns were fitted to acrylic replicas of ideal crown preparations and were divided equally into 3 test groups: group 1--crowns were crimped only (no cement used); group 2--crowns were cemented only; and group 3--crowns were crimped and cemented to the acrylic replicas. An Instron machine recorded the amount of force necessary to dislodge the crowns and the results were statistically analyzed using 2-way ANOVA and Tukey honestly significant difference (HSD) test. RESULTS: Group 3 was statistically more retentive than groups 1 and 2. Group 2 was statistically more retentive than group 1 (P < .001). In group 1, Unitek crowns were statistically more retentive than the veneered crowns (P < .05). In group 2, NuSmile crowns showed statistically less retention values than all other crowns (P < .05). In group 3,Kinder Krown crowns showed statistically better retention rates than all other brands (P < .05). CONCLUSIONS: Significantly higher retention values were obtained for all brands tested when crimping and cement were combined. The crowns with veneer facings were significantly more retentive than the nonveneered ones when cement and crimping were combined. PMID- 12889707 TI - The prevalence of periodontal-related changes in adolescents with asthma: results of the Third Annual National Health and Nutrition Examination Survey. AB - PURPOSE: The purpose of this study was to explore the association between asthma and periodontal disease in adolescents using oral examination and health interview data from the Third National Health and Nutrition Examination Survey (NHANES III) 1988-1994. METHODS: The study population comprised 1,596 adolescents 13 to 17 years of age: 253 (16%) asthmatics and 1,358 (84%) nonasthmatic controls who were examined for bleeding on probing (BOP), subgingival calculus (SBC), supragingival calculus (SPC), probing depth greater than or equal to 3 mm (PD), and loss of periodontal attachment greater than or equal to 2 mm (LPA). The authors fitted separate multivariate GEE Poisson regression models adjusting for parents' income, gender, race, exposure to potentially xerogenic drugs (antihistamines, corticosteroids, and inhalers), tobacco exposure, and dental examination within the past year. RESULTS: None of the periodontal measures was associated with asthma severity or with the use of antiasthmatic drugs. However, several covariates had statistically significant odds ratios (P < .05). CONCLUSIONS: There was no evidence to support the association between asthma and periodontal health in the adolescent population. Since the findings may be due to the inherent limitations of cross-sectional studies, the lack of knowledge about the daily dose of antiasthmatic medication, and the level of-compliance with the therapeutic regimen, future studies should be longitudinal and monitor medication use. PMID- 12889708 TI - War and rebuilding: what can we expect in the aftermath of the war in Iraq? PMID- 12889709 TI - Botanical briefs: Capsicum peppers. AB - Capsicum peppers are widespread, both in and out of the kitchen, both nationally and internationally. They contribute to burning and stinging of hands, mouth, and eyes in conjunction with food preparation and ingestion that is often unreported to the medical community. The literature is not abundant on the subject and often is confusing. This article provides brief yet detailed information about this important plant. PMID- 12889710 TI - What is your diagnosis? Targetoid hemosiderotic hemangioma. PMID- 12889711 TI - Cutaneous T-cell lymphoma in a patient with neurofibromatosis type 1. AB - A variety of neoplasms may occur in association with neurofibromatosis type 1 (NF1). We describe a patient with NF1 and mycosis fungoides. Recommendations for the initial and long-term evaluations of patients with neurofibromatosis are presented. PMID- 12889712 TI - Bullous eruption: a manifestation of lupus erythematosus. AB - Bullous lupus erythematosus (BSLE) is a rare subset of systemic lupus erythematosus (SLE), often associated with autoimmunity to type VII collagen. Generally, patients with BSLE meet the criteria for SLE as defined by the American College of Rheumatology. We present a case of a 17-year-old adolescent girl who presented with a vesiculobullous eruption without detectable type VII collagen antibodies and without full criteria for SLE. Differential staining was characteristic for lupus erythematosus (LE), suggesting her eruption is related to LE. We review the spectrum of bullous disease in patients with LE and discuss the pathogenesis and histology of these eruptions, as well as current therapeutic options. PMID- 12889713 TI - Diseases encountered during war and rebuilding: lessons from past conflicts. AB - As we face the task of helping to rebuild Iraq, it is worthwhile to review our experience with past humanitarian aid missions and past conflicts. These efforts serve as models for the types of problems we may encounter today. It also is critical to review available data concerning the unique aspects of cutaneous diseases in that part of the world. PMID- 12889714 TI - Skin reaction following immunization with smallpox vaccine: a personal perspective. AB - Concerns about the possibility of bioterrorism or unconventional weaponry using the variola virus have emerged following the events of September 11, 2001. Immunization of emergency personnel in Israel against smallpox started in September 2002. This case report describes the skin reaction following immunization with vaccinia virus. Physicians should be aware of the variations at the site of the inoculation. PMID- 12889715 TI - Necrobiotic xanthogranuloma associated with a benign monoclonal gammopathy. AB - Necrobiotic xanthogranuloma (NXG) is a disorder characterized by indurated, yellow-red nodules or plaques, primarily involving the face and, less frequently, the trunk and extremities. NXG may be associated with paraproteinemia, multiple myeloma, and hypertension. Histologically, xanthogranulomatous features with hyaline necrosis or necrobiosis are present. No first-line treatment has been established. This disease is a chronic process, and a patient's prognosis depends on the degree of extracutaneous involvement and the presence of visceral malignancies. We describe a patient with typical cutaneous and histologic findings of NXG with an associated monoclonal gammopathy. PMID- 12889716 TI - Enoxaparin-induced generalized exanthem. AB - Adverse cutaneous reactions to low molecular weight heparin (LMWH) are uncommon in the literature. They are usually localized reactions such as ecchymoses, erythematous plaques, and nodules. Only one case of a generalized maculopapular rash with enoxaparin has been reported in Europe. To our knowledge, this is the first case reported in the English literature of a generalized exanthem due to subcutaneous injection of enoxaparin. PMID- 12889717 TI - Lichen planopilaris presenting as truncal alopecia: a case presentation and review of the literature. AB - Lichen planopilaris is an inflammatory condition of unknown etiology, characterized by violaceous folliculocentric papules and hair loss. Some clinicians consider lichen planopilaris to be a variant of lichen planus, but others believe it to be a separate disease entity. Many treatment modalities have been utilized, with varying degrees of success. We describe the case of a 63-year old man who presented with widespread alopecia of the trunk and extremities and was subsequently diagnosed with lichen planopilaris. PMID- 12889718 TI - Efficacy and safety of a new triple-combination agent for the treatment of facial melasma. AB - Treatment of melasma, a hyperpigmentation disorder, remains a challenge. The primary objective of two 8-week, multicenter, randomized, investigator-blind studies was to compare the efficacy and safety of a hydrophilic cream formulation containing tretinoin 0.05%, hydroquinone 4.0%, and fluocinolone acetonide 0.01% (RA+HQ+FA) with the dual-combination agents tretinoin plus hydroquinone (RA+HQ), tretinoin plus fluocinolone acetonide (RA+FA), and hydroquinone plus fluocinolone acetonide (HQ+FA). All agents had the same drug concentration and vehicle. A total of 641 adult patients, predominantly female, with moderate to severe melasma and Fitzpatrick skin types I through IV, were randomized to the various treatment groups. Due to the similarity of the study designs, the results of the 2 studies were combined and are reported here. The primary efficacy analysis involved the proportion of intent-to-treat patients in each treatment group whose condition had completely cleared by week 8. The results of the combined clinical trials demonstrated that significantly more of the patients treated with RA+HQ+FA (26.1%) experienced complete clearing compared with the other treatment groups (4.6%) at the end of week 8 (P<.0001). In addition, at week 8, a 75% reduction in melasma/pigmentation was observed in more than 70% of patients treated with RA+HQ+FA compared with 30% in patients treated with the dual-combination agents. The most common adverse reactions seen with all treatment groups were erythema, skin peeling, burning, and/or stinging sensation. The majority of treatment related adverse events were of mild severity. PMID- 12889719 TI - Cumulative irritation comparison of adapalene gel and solution with 2 tazarotene gels and 3 tretinoin formulations. AB - Forty-two subjects with normal skin were enrolled in a single-center study to assess the cumulative irritancy potential of adapalene (Differin gel 0.1% and Differin solution 0.1%) compared with tazarotene (Tazorac gels 0.05% and 0.1%), tretinoin (Retin-A Micro gel 0.1%, Avita cream 0.025%, and Avita gel 0.025%), and white petrolatum (negative control). All test materials were applied randomly, under occlusion, to sites located on either side of the midline--the mid thoracic area of the subjects' backs. All patches were applied daily, Monday through Friday, to the same sites, unless the degree of reaction to a test product or adhesive necessitated removal (grade 3). Thirty-eight of the 42 subjects (90.5%) completed the study. Thirty-four of those 38 subjects (89.5%) had to discontinue using both tazarotene concentrations due to intolerance. Patch discontinuations for the remaining test materials were as follows: 7 subjects discontinued use of tretinoin microsphere gel 0.1%, 3 discontinued tretinoin cream 0.025%, 1 discontinued tretinoin gel 0.025%, and 1 discontinued adapalene gel 0.1%. None of the subjects discontinued use of the white petrolatum or the adapalene solution 0.1%. Adapalene gel and solution 0.1% were statistically (P<.01) less irritating than both tazarotene gels 0.1% and 0.05%, tretinoin microsphere gel 0.1%, and tretinoin gel 0.025%, and they were not statistically different from tretinoin gel 0.025%. PMID- 12889720 TI - Nursing sickness in the mink--a metabolic mystery or a familiar foe? AB - Nursing sickness, the largest single cause of mortality in adult female mink (Mustela vison), is an example of a metabolic disorder, which develops when the demands for lactation require extensive mobilization of body energy reserves. The condition is characterized by progressive weight loss, emaciation, and dehydration with high concentrations of glucose and insulin in the blood. Morbidity due to nursing sickness can be as high as 15% with mortality around 8%, but the incidence is known to vary from year to year. Stress has been shown to trigger the onset of the disease and old females and females with large litters are most often affected. Increasing demand for gluconeogenesis from amino acids due to heavy milk production may be a predisposing factor. Glucose metabolism is inextricably linked to that of protein and fats. In obesity (or lipodystrophy), the ability of adipose tissue to buffer the daily influx of nutrients is overwhelmed (or absent), interfering with insulin-mediated glucose disposal and leading to insulin resistance. Polyunsaturated fatty acids of the n-3 family play an important role in modulating insulin signalling and glucose uptake by peripheral tissue. The increasing demand on these fatty acids for milk fat synthesis towards late lactation may result in deficiency in the lactating female, thus impairing glucose disposal. It is suggested that the underlying cause of mink nursing sickness is the development of acquired insulin resistance with 3 contributing key elements: obesity (or lipodystrophy), n-3 fatty acid deficiency, and high protein oxidation rate. It is recommended that mink breeder females be kept in moderate body condition during fall and winter to avoid fattening or emaciation. A dietary n-3 fatty acid supplement during the lactation period may be beneficial for improved glycemic control. Lowering of dietary protein reduces (oxidative) stress and improves water balance in the nursing females and may, therefore, prevent the development and help in the management of nursing sickness. It is also surmised that other, thus far unexplained, metabolic disorders seen in male and female mink may be related to acquired insulin resistance. PMID- 12889721 TI - Accuracy of pulse oximetry and capnography in healthy and compromised horses during spontaneous and controlled ventilation. AB - The objective of this prospective clinical study was to evaluate the accuracy of pulse oximetry and capnography in healthy and compromised horses during general anesthesia with spontaneous and controlled ventilation. Horses anesthetized in a dorsal recumbency position for arthroscopy (n = 20) or colic surgery (n = 16) were instrumented with an earlobe probe from the pulse oximeter positioned on the tip of the tongue and a sample line inserted at the Y-piece for capnography. The horses were allowed to breathe spontaneously (SV) for the first 20 min after induction, and thereafter ventilation was controlled (IPPV). Arterial blood, for blood gas analysis, was drawn 20 min after induction and 20 min after IPPV was started. Relationships between oxygen saturation as determined by pulse oximetry (SpO2), arterial oxygen saturation (SaO2), arterial carbon dioxide partial pressure (PaCO2), and end tidal carbon dioxide (P(et)CO2), several physiological variables, and the accuracy of pulse oximetry and capnography, were evaluated by Bland-Altman or regression analysis. In the present study, both SpO2 and P(et)CO2 provided a relatively poor indication of SaO2 and PaCO2, respectively, in both healthy and compromised horses, especially during SV. A difference in heart rate obtained by pulse oximetry, ECG, or palpation is significantly correlated with any pulse oximeter inaccuracy. If blood gas analysis is not available, ventilation to P(et)CO2 of 35 to 45 mmHg should maintain the PaCO2 within a normal range. However, especially in compromised horses, it should never substitute blood gas analysis. PMID- 12889722 TI - Development and analytic validation of an enzyme-linked immunosorbent assay for the measurement of canine pancreatic lipase immunoreactivity in serum. AB - Recently, a radioimmunoassay (RIA) for measurement of canine pancreatic lipase immunoreactivity (cPLI) in serum was developed and validated. However, RIAs require frequent use of radioactive materials. Therefore, the goal of this project was to develop and validate an enzyme-linked immunosorbent assay (ELISA) for cPLI. After purifying cPL, we developed and purified antiserum against cPL in rabbits. The purified antibody was bound to microtitre plates and used to capture antigen. A portion of the purified antibody was biotinylated and used to identify the captured antigen. Streptavidin labelled with horseradish peroxidase and a horseradish peroxidase substrate were used for detection. The assay was validated by determination of sensitivity, working range, linearity, accuracy, precision, and reproducibility. The reference interval for serum cPLI was determined by the central 95th percentile in 74 clinically healthy dogs: 2.2 to 102.1 microg/L. The sensitivity and the upper limit of the working range were 0.1 and 999.2 microg/L, respectively. The ratios of observed to expected values for dilutional parallelism for 6 serum samples ranged from 0.0 to 148.8%; the ratios for spiking recovery for 4 serum samples ranged from 90.4 to 112.6%, assuming 55% recovery of the cPL. Coefficients of variation for intra- and interassay variability for 6 different serum samples were 2.4, 3.4, 4.1, 5.8, 7.4, and 10.0% and 5.9, 7.7, 11.6, 13.9, 23.5, and 46.2%, respectively. We conclude that the ELISA described here is sufficiently sensitive, linear, accurate, precise, and reproducible for clinical application. Evaluation of its clinical usefulness for the diagnosis of exocrine pancreatic disorders in dogs is under way. PMID- 12889723 TI - Use of a 13C-aminopyrine blood test: first clinical impressions. AB - The purpose of this study was to collect initial data to determine the potential clinical usefulness of a 13C-aminopyrine demethylation blood test, and whether additional clinical investigation is warranted. Six dogs, initially suspected of having hepatic disease based on their history, physical examination, imaging studies, general laboratory parameters, or any combination of the above, were enrolled in the study. A baseline blood sample was collected, 2 mg/kg 13C aminopyrine was administered intravenously, and another blood sample was collected 45 min afterwards. Carbon dioxide was extracted from the blood samples and analyzed using fractional mass spectrometry. Results from the 13C-aminopyrine demethylation blood test were compared to clinical data and histologic findings. Intravenous administration of 13C-aminopyrine leads to a decrease in the percent dose of 13C recovered from dogs with histologically confirmed liver disease. Based on our results, a full-scale investigation of the potential clinical usefulness of a 13C-aminopyrine demethylation blood test for assessment of hepatic function in dogs is warranted. PMID- 12889724 TI - Enzyme activity in bovine cervical mucus during spontaneous and induced estrus. AB - The purpose of the present research was to compare the enzyme activity of alkaline phosphatase (ALP), lactate dehydrogenase (LDH), alpha-amylase, alpha manosidase, beta-N-acetyloglucosaminidase, beta-glucuronidase, and beta galactosidase in the cervical mucus of cows during spontaneous and induced estrus. Friesian cows (n = 106) were assigned to 4 groups: 1) no treatment; 2) progesterone releasing intervaginal device (PRID) for 12 days plus pregnant mare serum gonadotrophin (PMSG) at the removal of the PRID; 3) PGF2alpha 2 doses 11 days apart; and 4) PRID for 7 days plus PGF2alpha 1 dose, 24 hours before removal of the PRID. Fourteen cows were excluded from the trial because of an inadequate quantity of cervical mucus collected or a lost PRID. The cows from the 3 induced estrus groups were artificially inseminated (AI) twice, while those with spontaneous estrus received only a single AI. Cervical mucus samples were collected from all cows 5 to 30 min before the first AI. The results are summarized as follows: 1) ALP and alpha-amylase activity for spontaneous estrus were similar to those for induced estrus; 2) LDH activity levels during spontaneous estrus were significantly lower (P < 0.001) than that in the P4 and P4+PGF2alpha induced estrus groups; and 3) glycosidases' activity was significantly lower (P < 0.001) in the spontaneous estrus group than that in the induced estrous groups. In conclusion, the activity of most enzymes in the cervical mucus of cows, in the present study, was significantly different between the spontaneous and the induced estrus groups. PMID- 12889725 TI - Effect on abortion of feeding Korean pine needles to pregnant Korean native cows. AB - We studied the frequency and timing of abortion and the serum levels of 17beta estradiol and progesterone in Korean native cows fed pine needles during pregnancy. Fifteen pregnant cows were randomly assigned to groups of 5. The control group was fed a concentrate and rice straw, and the other 2 groups were fed, in addition, either 1.3 or 2.7 kg (dry weight) of Korean pine needles daily, starting at an average of 91 d of gestation and continuing until 245 d of gestation. The health status of the dams and the viability of the fetuses were ascertained by rectal palpation and ultrasound scanning during pregnancy. Simultaneously, blood samples were collected for analysis of serum 17beta estradiol and progesterone. Two abortions in mid-pregnancy (at 126 and 150 d of gestation) occurred in the group ingesting the higher daily amount of pine needles. Premature parturition occurred at 259 d of gestation in this group and at 241 and 252 d of gestation in the group ingesting the lower daily amount of pine needles. The serum 17beta-estradiol concentration was significantly higher (P < 0.05) at 4 mo of gestation and the serum progesterone level significantly lower (P < 0.05) at 8 mo of gestation in the group ingesting more pine needles daily than in either of the other 2 groups. These results suggest that the ingestion of pine needles may play a role in abortion in Korean native cows by increasing the serum 17beta-estradiol concentration and decreasing the serum progesterone concentration. PMID- 12889726 TI - Survival of porcine reproductive and respiratory syndrome virus in houseflies. AB - The objectives of the study were to determine the duration of porcine reproductive and respiratory syndrome virus (PRRSV) survival in houseflies (Musca domestica Linnaeus) following feeding on an infected pig, and to determine whether the virus was present on the exterior surface or within the internal viscera of the fly. A total of 210 laboratory-colonized houseflies were allowed to feed to repletion on a pig, experimentally infected with PRRSV on day 7 postinoculation, and then maintained alive under laboratory conditions (27 degrees C). Two subsets (A and B) of 30 flies were collected at each of the following sampling points; 0, 6, and 12 hours post feeding (pf). Subset A contained an extra group of 30 flies collected at 24 hours pf due to the availability of extra flies. Flies in subset A were processed as whole fly homogenates, while the exterior surface washes and digestive organs were collected from flies in subset B. Whole fly homogenates, collected at 0, 6, and 12 hours pf, were positive by both polymerase chain reaction (PCR) and swine bioassay. Digestive organs, collected at 0 and 12 hours pf, were positive by PCR and swine bioassay. The PRRSV RNA was detected by PCR from the exterior surface wash of subset B flies collected at 0, 6, and 12 hours pf; however, only the subset collected at 0 hour pf was swine bioassay-positive. This study indicates that infectious PRRSV can survive within the intestinal tract of houseflies for up to 12 hours following feeding on an infected pig, but only for a short period on the exterior surface of the flies. PMID- 12889727 TI - Effects of diet formulations containing proteins from different sources on intestinal colonization by Campylobacter jejuni in broiler chickens. AB - The objective of this study was to compare the effects of 3 diet formulations containing different protein sources (animal, plant, and a combination of animal and plant) on the colonization of Campylobacter jejuni in the gastrointestinal tract of broiler chickens. A freshly isolated strain of C. jejuni (biotype IV, serotype HS O:21, O:29, HL untypable) from a broiler chicken was used to infect 3 day-old chicks that had been free of C. jejuni; 0.5 mL of an inoculum containing 10(8) colony-forming units was administered orally. Shedding of the organism was studied, and C. jejuni in the ceca, jejuni, and crop were enumerated by quantitative culture. The isolates recovered from the birds during the study period of 35 d were characterized and confirmed as C. jejuni by the use of standard methods and underwent biotyping, serotyping, antimicrobial susceptibility testing by disk diffusion and the E-test, and flagellin gene typing. A cyclical pattern of shedding of C. jejuni was observed in all the birds. Colonization was highest in the ceca. The ceca of birds receiving plant protein-based feed had significantly less colonization then the ceca of birds receiving the other types of feed, whereas the differences in colonization of the jejuni and crops were not significant. Characterization by biotyping, serotyping, and flagellin gene typing showed that 95% of the recovered isolates were identical to the strain used for infecting the chicks. However, with the Lior-HL typing scheme, 74% of the recovered isolates were HL untypable. Antimicrobial resistance testing did not reveal significant differences between the infecting strain and the recovered isolates among the different feed groups. PMID- 12889728 TI - Diagnostic testing patterns of natural Mycobacterium paratuberculosis infection in pygmy goats. AB - Thirteen pygmy goats (Capra hircus) from a herd naturally infected with Mycobacterium avium ss. paratuberculosis (MPTB) were monitored with 4 diagnostic assays for 2 to 15 mo. Cellular and humoral immune responses to the infection were assessed with assays of gamma interferon (IFNgamma), serum antibody [enzyme linked immunosorbent assay (ELISA) and agar gel diffusion (AGID)], and radiometric fecal culture. Microscopic examination and radiometric culture of tissue from 12 sites were performed at necropsy. Goats were considered infected if MPTB was isolated from any tissue sample collected at necropsy. Mycobacterial isolates were confirmed as MPTB with an IS900 polymerase chain reaction assay. Ten goats whose antemortem tests indicated infection carried heavy organism burdens at necropsy, both within and beyond the gastrointestinal system. False negative ELISA, AGID, and/or culture results were obtained in 5 of the 10 confirmed cases during the study period. In 3 goats with sporadic fecal shedding of MPTB or detectable IFNgamma response, or both, no abnormalities were detected at necropsy and no MPTB was isolated from the tissue samples; the antemortem fecal-culture and IFNgamma results were thus considered false-positive. Diagnosticians should be alert to the possibility of both false-positive and false-negative test results for Johne's disease in goats. False-positive fecal culture results may occur when a high prevalence of infection exists in the herd and the premises are likely to be heavily contaminated. The diverse antemortem testing patterns seen in these goats underscore the importance of using varied diagnostic assays serially or in parallel to increase the likelihood of identifying all infected goats. PMID- 12889729 TI - Comparison of bacterial enriched-broth culture, enzyme linked immunosorbent assay, and broth culture-polymerase chain reaction techniques for identifying asymptomatic infections with Salmonella in swine. AB - A polymerase chain reaction (PCR) assay was combined with a broth-culture enrichment system to detect Salmonella shed in feces from subclinically infected swine. The effectiveness of the broth culture-polymerase chain reaction (BC-PCR) assay to identify pigs shedding Salmonella in feces was compared with a microbiological culture and a commercial enzyme linked immunosorbent assay (ELISA) kit to detect Salmonella-specific serum antibody. A total of 67 pigs were tested by each of the 3 methodologies. Forty-one pigs tested positive for Salmonella by BC-PCR and ELISA identified 6 positives and 23 suspicious samples. It was shown that the BC-PCR assay is a rapid diagnostic tool for detecting of Salmonella shed by asymptomatic swine compared with current diagnostic technologies. PMID- 12889730 TI - Comparison of experimental models for Streptococcus suis infection of conventional pigs. AB - Four different experimental models for Streptococcus suis-induced disease were compared to find a model that closely mimics naturally occurring disease in conventional pigs. Fourteen, 2-week old pigs free of S. suis type 2 were used in 2 experiments. In experiment 1, 3 pigs were inoculated intravenously (IV) and 3 pigs intranasally (IN) with S. suis. Two out of 3 of the IV-inoculated pigs exhibited signs of severe central nervous system disease (CNS) and were euthanized. Streptococcus suis type 2 was isolated from whole blood, joints, and serosal surfaces of both pigs. No clinical signs and no growth of S. suis were detected in the IN-inoculated pigs. In experiment 2, 4 pigs were inoculated IV and another 4 were inoculated IN with the same isolate as in experiment 1. One hour before inoculation the IN-inoculated pigs were given 5 mL of 1% acetic acid intranasally (IN-AA). All the IV-inoculated pigs showed CNS disease and lameness, and 2 of the pigs became severely affected and were euthanized. All the IN-AA inoculated pigs exhibited roughened hair coats and 2 pigs developed severe CNS disease and were euthanized. Streptococcus suis was isolated from the joints and blood of 3 pigs in the IV-inoculated group. Streptococcus suis was isolated from blood of 2 pigs, meninges of 3 pigs, and joints of 1 pig in the IN-AA inoculated group. Natural exposure to S. suis most likely occurs by the intranasal route. The IN-AA model should serve as a good model for S. suis-induced disease, because the natural route of exposure is intranasal and the IN-AA model was effective in inducing disease that mimics what is observed in the field. PMID- 12889731 TI - Levels of nicotinamide adenine dinucleotide in extracellular body fluids of pigs may be growth-limiting for Actinobacillus pleuropneumoniae and Haemophilus parasuis. AB - During infection, nutrient deprivation can alter bacterial phenotype. This, in turn, may have implications for pathogenesis and prophylaxis. Actinobacillus pleuropneumoniae (biotype 1) and Haemophilus parasuis, respiratory tract pathogens of swine, are both V-factor-dependent. The concentrations of V factor in the extracellular fluids of pigs are unknown and may limit the growth of these bacteria in vivo. The aim of this study was to determine the concentrations of nicotinamide adenine dinucleotide (NAD) in select porcine body fluids and to compare the availability of NAD in vivo with the affinities of the organisms for this compound. Levels in plasma, tissue fluids (peritoneal, pleural, synovial, and cerebrospinal), and laryngeal, tracheal, and lung washings were determined with an enzymatic cycling assay. We concluded that, although the NAD supply in the respiratory tract is probably not growth-limiting, it may become limiting if the organisms are disseminated. PMID- 12889732 TI - Search for Lawsonia intracellularis and Bilophila wadsworthia in malabsorption diseased chickens. AB - Proliferative enteropathy is an important enteric disease caused by Lawsonia intracellularis. A wide range of host species can be infected by the same bacterium, yet the clinico-pathologic features among these hosts remains almost identical. The disease has been recognized regularly among ratites, but not in other avian families, such as galliforms, even though these suffer uncharacterized enteric conditions. Fresh ileum-colon contents were obtained from 228, 3- to 8-week-old chickens with enteric disease, kept at 14 large commercial farms in the southern USA. DNA was extracted from each sample and subjected to polymerase chain reactions (PCR) with primers specific to eubacterial DNA, L. intracellularis, and Bilophila wadsworthia. All chicken samples were positive for eubacterial DNA, 29 chickens (13%) were positive for B. wadsworthia DNA, and none were positive for L. intracellularis DNA. Given the ubiquitous nature of L. intracellularis, we consider it likely that some avian families do not carry the necessary mechanism for L. intracellularis viability. Bilophila wadsworthia appears to be a consistent member of the colonic flora of some host animals. Neither bacterium appears to be associated with malabsorption syndromes in chickens. PMID- 12889733 TI - Expression of beta2-integrin on monocytes and blood polymorphonuclear leukocytes in the periparturient period in dairy cows. AB - The hypothesis that an altered expression of CD11/CD18 on bovine circulating monocytes, polymorphonuclear leukocytes (PMN), or both, contributes to an increased mastitis susceptibility in periparturient cows was tested. Expression of CD18 and CD11a, -b, -c on bovine monocytes and PMN were assessed in 8 Friesian Holstein cows by flow cytometry from 2 wk before calving to 5 wk after calving. Minor changes in adhesion molecule expression levels were detected throughout the experimental period. Compared with PMN, monocytes exhibited an expression level that was similar for CD18, higher for CD11a and CD11c, but lower for CD11b. Differences in density may reflect the relative importance of these adhesion molecules on both leukocyte types. In this study, the decreased number of milk resident macrophages and PMN observed during the periparturient period could not be attributed to changes of CD11/CD18 levels on circulating leukocytes. PMID- 12889734 TI - Effects of aminoglycoside antibiotics on renal antioxidants, malondialdehyde levels, and some serum biochemical parameters. AB - Effects of amikacin, gentamicin, kanamycin, and streptomycin on renal tissue superoxide dismutase, glutathione peroxidase, glutathione and malondialdehyde, serum creatinine, potassium, sodium, total protein, glucose, uric acid, and total bilirubin levels were investigated. All aminoglycoside antibiotics decreased renal tissue glutathione levels. PMID- 12889735 TI - Toxicity patterns of cytotoxic drugs. AB - Toxicity is a major concern for anticancer drugs. These compounds present a narrow therapeutic index, with a small difference between the dose required for an antitumor effect and that responsible for unacceptable toxicity. Their recommended doses are determined according to the toxicity endpoint. Moreover, toxicity is observed earlier than the therapeutic effect, so, toxic effects represent a major endpoint for pharmacodynamic studies of cytotoxic drugs. Knowledge of toxicity patterns and main factors of toxicity of anticancer drugs is required before modeling data of these studies. Hematological toxicities represent the main toxicity of the cytotoxic. However, non-hematological toxicities have become more important than hematological toxicities as pharmacodynamic endpoints in some circumstances such as high-dose chemotherapy associated with bone marrow transplantation. This paper will describe the main toxicity of the cytotoxic drugs, and its factors of both inter- and intra-patient variability. The toxicity pattern of topotecan will be examined as an example. Knowledge of the toxicity pattern of a drug constitutes a prerequirement before modeling its pharmacodynamics. PMID- 12889737 TI - Noninvasive methods to study drug distribution. AB - Positron emission tomography (PET) and nuclear magnetic resonance spectroscopy (MRS) are two techniques that allow the noninvasive monitoring of drug distribution in living systems (humans, animals), and dynamic contrast-enhanced magnetic resonance imaging (dMRI) provides noninvasive physiological information relevant for drug distribution. PET yields series of cross-sectional images that can be used to monitor the absolute radioactivity concentrations in tissues pixel by-pixel, but does not allow direct identification of each of the products present. MRS produces spectra showing changes in the concentration of both the parent drug and of the metabolites separately for a sensitive volume, but does not provide a simple means for measuring absolute concentrations. dMRI, which measures the changes in the rates of relaxation of water, proportional to the concentrations of the contrast agent (usually Gd-DTPA), readily allows the determination of functional changes in cross-sectional images down to a pixel-by pixel level. All of these methods are of special interest to evaluate the amounts of drug that can reach the target tissue, penetrate it, remain present at such targets for a sufficient length of time, and how they are metabolized at the target site. Such information may be of particular interest in the study of solid malignant tumors and may become very relevant for determining better treatment strategies. This article presents examples of successful studies of tissue pharmacokinetics with MRS and dMRI. The following article is devoted to PET. PMID- 12889736 TI - Chemotherapy individualization. AB - The current practice of dosing patients with anticancer drugs based on body size, leads to a large degree of interpatient variation in clinical outcome following standard doses of chemotherapy. Some patients may fail to respond to treatment, whilst others experience unacceptable side effects. Recent studies have identified more rational approaches to drug dosing, based on patient characteristics such as renal function, pharmacogenetic factors, and drug metabolizing activity. These can be used together with therapeutic drug monitoring and adaptive dosing to achieve a targeted systemic drug exposure in each patient, which may lead to more consistent clinical outcomes in patients receiving comparable chemotherapy dosing regimens. The purpose of this review is to present some approaches to chemotherapy individualization, examples of how this might be applied, and speculation as to how recent advances in pharmacogenetics may lead to further dose-optimization. Whilst it is hoped that the design of new agents, targeted to specific genes involved in oncogenesis, will lead to increased success in the treatment of cancer patients, it is essential that the drugs currently available are used to their maximum potential. PMID- 12889738 TI - Use of positron emission tomography in anticancer drug development. AB - Positron emission tomography (PET) is increasingly being used in anticancer drug development. The technique is applicable to studies of drug delivery, and where specific probes are available, to provide pharmacodynamic readouts noninvasively in patients. Mathematical modeling of the imaging data enhances the quality of information that is obtained from such studies. This section provides a review of the PET methodologies that have been used for the development of new cancer therapies. Other than imaging of radiolabeled drugs, PET modeling has found extensive application in studies with 2-[11C]thymidine, [18F]fluorodeoxyglucose, H2(15)O, C15O, and receptor ligands. PMID- 12889739 TI - Mechanistic models for myelosuppression. AB - As myelosuppression is the dose-limiting toxicity for most chemotherapeutic drugs, modelers attempt to find relationships between drug and toxicity to optimize treatment. Mechanistic models, i.e. models based on physiology and pharmacology, are preferable over empirical models, as prior information can be utilized and as they generally are more reliable for extrapolations. To account for different dosing-regimens and possible schedule-dependent effects, the whole concentration-time profile should be used as input into the pharmacokinetic pharmacodynamic model. It is also of importance to model the whole time course of myelosuppression to be able to predict both the degree and duration of toxicity as well as consecutive courses of therapy. A handful of (semi)-mechanistic pharmacokinetic-pharmacodynamic models with the above properties have been developed and are reviewed. Ideally, a model of myelosuppression should separate drug-specific parameters from system related parameters to be applicable across drugs and useful under different clinical settings. Introduction of mechanistic models of myelosuppression in the design and evaluation of clinical trials can guide in the decision of optimal sampling times, contribute to knowledge of optimal doses and treatment regimens at an earlier time point and identify sub groups of patients at a high risk of myelosuppression. PMID- 12889740 TI - Clinical pharmacokinetic/pharmacodynamic and physiologically based pharmacokinetic modeling in new drug development: the capecitabine experience. AB - Preclinical studies, along with Phase I, II, and III clinical trials demonstrate the pharmacokinetics, pharmacodynamics, safety and efficacy of a new drug under well controlled circumstances in relatively homogeneous populations. However, these types of studies generally do not answer important questions about variability in specific factors that predict pharmacokinetic and pharmacodynamic (PKPD) activity, in turn affecting safety and efficacy. Semi-physiological and clinical PKPD modeling and simulation offer the possibility of utilizing data obtained in the laboratory and the clinic to make accurate characterizations and predictions of PKPD activity in the target population, based on variability in predictive factors. Capecitabine is an orally administered pro-drug of 5 fluorouracil (5-FU), designed to exploit tissue-specific differences in metabolic enzyme activities in order to enhance efficacy and safety. It undergoes extensive metabolism in multiple physiologic compartments, and presents particular challenges for predicting pharmacokinetic and pharmacodynamic activity in humans. Clinical and physiologically based pharmacokinetic (PBPK) and pharmacodynamic models were developed to characterize the activity of capecitabine and its metabolites, and the clinical consequences under varying physiological conditions such as creatinine clearance or activity of key metabolic enzymes. The results of the modeling investigations were consistent with capecitabine's rational design as a triple pro-drug of 5-FU. This paper reviews and discusses the PKPD and PBPK modeling approaches used in capecitabine development to provide a more thorough understanding of what the key predictors of its PBPK activity are, and how variability in these predictors may affect its PKPD, and ultimately, clinical outcomes. PMID- 12889741 TI - Pharmacokinetic-pharmacodynamic guided trial design in oncology. AB - The application of pharmacokinetic (PK) and pharmacodynamic (PD) modeling in drug development has emerged during the past decades and it is has been suggested that the investigation of PK-PD relationships during drug development may facilitate and optimize the design of subsequent clinical development. Especially in oncology, well designed PK-PD modeling could be extremely useful as anticancer agents usually have a very narrow therapeutic index. This paper describes the application of the current insights in the use of PK-PD modeling to the design of clinical trials in oncology. The application of PK-PD modeling in each separate stage of (pre)clinical drug development of anticancer agents is discussed. The implementation of this approach is illustrated with the clinical development of docetaxel. PMID- 12889743 TI - The enzymology of combinatorial biosynthesis. AB - Combinatorial biosynthesis involves the genetic manipulation of natural product biosynthetic enzymes to produce potential new drug candidates that would otherwise be difficult to obtain. In either a theoretical or practical sense, the number of combinations possible from different types of natural product pathways ranges widely. Enzymes that have been the most amenable to this technology synthesize the polyketides, nonribosomal peptides, and hybrids of the two. The number of polyketide or peptide natural products theoretically possible is huge, but considerable work remains before these large numbers can be realized. Nevertheless, many analogs have been created by this technology, providing useful structure-activity relationship data and leading to a few compounds that may reach the clinic in the next few years. In this review the focus is on recent advances in our understanding of how different enzymes for natural product biosynthesis can be used successfully in this technology. PMID- 12889742 TI - Clinical trial design for target specific anticancer agents. AB - Recently a large number of new anticancer agents targeting specifically one or more of the extracellular, transmembrane, or intracellular (but extranuclear) processes involved in malignant transformation of cells or carcinogenesis have been developed. These agents show target specificity, predominantly resulting in growth inhibition in tumor models and less frequently in tumor regression, acting in a cytostatic rather than a cytotoxic way. In addition, based on their specific mechanism of action, these target specific agents are expected to have a more favorable toxicity profile. In exploring new anticancer agents, phase I studies generally focus on toxicity and are primarily designed to describe dose limiting toxicity and to determine the maximum tolerated dose and the dose recommended for phase II studies. These phase II studies are subsequently performed in small groups of patients using the percentage tumor regression to screen for anticancer efficacy. Due to the anticipated low toxicity profile and the mainly growth inhibiting activity of target specific agents, the design of phase I and II studies involving these agents will have to be adapted in several ways. It is emphasized that, although it is helpful to distinguish cytotoxic from cytostatic anticancer agents, this dichotomy can be a simplification. In this paper, we will discuss important issues that will have to be faced when developing clinical trials with these agents and we will specifically translate this into the already known concepts of trial design exploring cytotoxic and cytostatic agents. PMID- 12889744 TI - Carbohydrates--the renewable raw materials of high biotechnological value. AB - Carbohydrates are the potential biomolecules derived from nature. Their molecular diversity has led to a bewildering variety of species, structures and characteristics all performing a large array of functions of great significance. Biologically they are vital as message (immunological) carriers, physiologically they are useful as energy (nutritional) reserves, and technologically they are needed for altering the texture and consistency (functional) of foods. Recent advances in glycobiology have opened up a new understanding of the role of sugars in biology and medicine. Noncellulosic beta-(1-3)-linked D-glucans, a group of polysaccharides found as constituents of fungi, algae, and higher plants, exhibit many interesting properties, depending on their molecular conformation. They are excellent 'biological response modifiers' and show significant immunomodulatory activities. They elicit a variety of host defense biological responses, for example, potent antitumor activity. On the other hand, the mixed-linkage ((1-3/1 4)-beta-linked) glucans are important constituents of cereal cell walls, where they perform properties of physiological importance, such as water holding capacity, porosity, and plasticity, which are useful at different stages of growth/development of plants. Of late, carbohydrate-based therapeutics are becoming the promise against many chronic diseases of today and tomorrow. Some of the characteristic features, structural attributes, functional significance, and applications of a selected few carbohydrate species are the subject matter of this review. PMID- 12889745 TI - Increased spending on health care: how much can the United States afford? AB - Perceptions of whether health care cost growth is affordable contribute greatly to pressures for health system reform. In this paper we develop a framework for thinking about affordability, concluding that a one-percentage-point gap between real per capita growth in health care costs and growth in GDP would be affordable through 2075. A two-percentage-point gap would only be affordable through 2039. In either case, the share of income growth devoted to health care would exceed historical norms. The value of care, which determines willingness to pay, and distributional issues are more important than our ability as a society to pay for care. PMID- 12889746 TI - Medical liability and patient safety. AB - Political debate over medical malpractice reform seldom takes meaningful account of its policy context, including the emerging science of patient safety. Instead, stakeholders on both sides use the rhetoric of patient safety to support entrenched positions on hardened proposals such as capping damages and limiting access to information about errors. Despite its deja vu quality, the current malpractice crisis can only be understood and addressed as the product of changes in the health care system since the last crisis nearly twenty years ago--changes that also informed the patient safety movement. Patient safety may therefore serve as a bridge between medical liability and health policy. PMID- 12889747 TI - The medical liability crisis of 2003: must we squander the chance to put patients first? AB - Medical liability reform should be aligned with a patient-centered, systems-based approach to preventing injury. Lessons learned about medical risk are now buried by the legal system, and communication about risk is haphazard among health care providers and across the interfaces of our legal, regulatory, and health care systems. Tort reform can be a vehicle for breaking down systemic barriers. Proposed reforms include (1) requiring disclosure of medical errors and restricting the use of information disclosed as evidence of guilt; (2) outlawing confidentiality agreements when malpractice cases are settled; (3) abolishing the National Practitioner Data Bank; and (4) establishing a national patient safety authority. PMID- 12889748 TI - Creating a safe environment. AB - William Sage provides an excellent rationalization for today's medical liability crisis. Patient safety and a safe environment are related concepts directed at helping to resolve this crisis. His paper helps to bring the complexities into perspective and suggests the need for an even better definition of "avoidable error" and for a righting of the tort system in the direction of the health care provider. I discuss here two ideas concerning injury compensation tables and medical liability specialty courts to help bring about this "righting." PMID- 12889749 TI - Federal health information policy: a case of arrested development. AB - Computerized patient records (CPRs) have reached a state of technical maturity that makes them an essential component of modern patient care. However, because uniform technical standards do not exist, CPRs constructed by different vendors do not convey clinical information easily from provider to provider. Moreover, unequal access to capital may mean a two-tier clinical information environment in the future. HIPAA, while important, did not anticipate the CPR revolution. New federal activism is required to assure not only interoperability of clinical data systems, but also that providers who lack capital and technical resources can make the needed digital conversion. PMID- 12889750 TI - Improving America's health care system by investing in information technology. AB - Large-scale investment in health care information technology (IT) infrastructure will not take place without leadership by the federal government. But how the federal government supports the financing of health care IT is critical. Health care IT development has multiple aspects, but it is fundamentally a problem of community infrastructure development. A policy approach that has had consistent success in financing our country's essential physical infrastructure in transportation and environmental protection will be well suited to fostering health care IT infrastructure as well. We propose the creation of a health care IT revolving loan fund program to invest public dollars in health care IT infrastructure projects through community-level nonprofit lending agencies. PMID- 12889751 TI - Realigning incentives in fee-for-service Medicare. AB - This paper proposes Medicare payment reform built on the fee-for-service system, with incentive payments to eligible provider organizations determined by their rate of increase in cost per patient compared to the overall growth rate in the community. By planning and monitoring how care patterns are altered to achieve greater efficiency, policy-makers can align the incentives of Medicare and the provider organization better than using either fee-for-service or capitation alone. This reform, unlike capitation, maintains Medicare's historical role as insurer and focuses providers on managing care. PMID- 12889752 TI - Medical schools and their applicants: an analysis. AB - Shortages of physicians have existed periodically throughout U.S. history. In response, medical school capacity has been increased, by either building new schools or expanding existing ones. Each strategy has encountered the obstacles of time, money, and applicants. If the United States chooses to increase its infrastructure for medical education again, these past experiences offer lessons that can be drawn upon. The most instructive ones are how long this process will take, how important public sponsorship and financing will be, and how much it will depend on antecedent dynamics within K-12 and baccalaureate education to assure an adequate flow of applicants, all of which makes the need to develop strategies for the future ever more pressing. PMID- 12889753 TI - Toil and trouble? Growing the physician supply. AB - Richard Cooper outlines several obstacles to increasing the supply of U.S. trained physicians. His analysis paints a bleak picture, but he may be underselling the possible benefits of technological advances that could increase medical school capacity and of increasing the volume of qualified applicants by broadening eligibility criteria. Another issue that would confront medical educators is that imbalances between supply and demand would not be uniform across states. This development could signal increased involvement from state policymakers, but public and private policymakers should take steps to assure that resources are funneled to where doctors are needed most and to exploit opportunities for innovative curriculum development. PMID- 12889754 TI - The future of medical education: a call for action. AB - The challenge of building new medical schools or expanding the class size of current ones is a lengthy and costly enterprise. That reality argues for the creation of a national vision about how to proceed and what to prioritize, a vision that could serve as a map for public and private officials considering further investment in medical education. A national commission funded by one or several health philanthropies is suggested as an expeditious vehicle to focus this discussion and move the debate forward. Instructive educational innovations in osteopathic and nursing education are suggested as germane to the deliberations of a national commission, as are creative means of student funding. PMID- 12889755 TI - A recurring theme: the need for minority physicians. AB - There is compelling evidence for the need to increase diversity within the physician workforce to ensure high-quality medical education, access to health care for the underserved, advances in research, and improved business performance. To have enough physicians to meet the future needs of the general public, as well as of minority citizens, we must recruit from diverse populations. The need for physicians, particularly under-represented minorities, will continue to grow. Addressing shortages requires inventive efforts to counter obstacles created by the anti-affirmative action movement, as well as strategies to encourage institutions to become more engaged in diversity efforts. PMID- 12889756 TI - The international medical graduate pipeline. AB - This paper presents information from the Educational Commission for Foreign Medical Graduates (ECFMG) on the certification and practice of international medical graduates (IMGs). IMGs constitute 25 percent of the U.S. physician workforce, and there is a vigorous pipeline of highly qualified candidates seeking graduate training in the United States. When considering how to address the potential physician shortfall described by Richard Cooper, policymakers will need to consider U.S. health care needs in the context of the intense international debate regarding global physician migration and its implications for the developing world. PMID- 12889757 TI - The physician workforce: a medical school dilemma. AB - Richard Cooper has advanced a projection of a sizable deficit in physicians in the United States, requiring the training of as many as 10,000 additional physicians annually by the year 2025. He questions the ability of U.S. medical schools to "fill the gap." This challenge presents another important dilemma for medical education. Could the applicant pool be enlarged sufficiently to increase qualified applicants? Would medical schools be able to accommodate this increase in students? The impact of these increases could be an ultimate decrease in the quality of health care and the production of physicians who are not current with research findings, particularly in the area of genetics. PMID- 12889758 TI - The curious conversion of Empire Blue Cross. AB - The for-profit conversion of Empire Blue Cross in New York challenges the case law and conventional policy wisdom that financial assets from formerly nonprofit organizations should be used to endow independent charitable foundations. The appropriation of Empire's assets by state government itself, and their subsequent deployment to subsidize health care institutions and repay political obligations, changes the conversion process from one that pits nonprofits against for-profits to one that pits private, nonprofit organizations against public-sector programs in the competition for new financial resources. PMID- 12889759 TI - Good science and the marketplace for drugs: a conversation with Jean-Pierre Garnier. Interview by John K. Iglehart. PMID- 12889760 TI - State initiatives on prescription drugs: creating a more functional market. AB - In response to unrelenting increases in prescription drug spending and use, many states are developing and implementing innovative policy solutions. The Reforming States Group (RSG), a nonpartisan organization of senior executive and legislative leaders from more than forty states and provinces formed in 1992, proposes a series of actions to improve the functioning of this market by introducing more explicit information on quality, effectiveness, and price and by experimental waiving of federal regulations. PMID- 12889761 TI - Focus on locus: evolution of Medicare's local coverage policy. AB - Medicare relies on a highly decentralized local-contractor structure that emerged as a political compromise in 1965. Decisions regarding Medicare's coverage of new procedures and technologies are an important part of the program. A national coverage process exists, but Medicare's local contractors develop most coverage policies. Although an intense debate surrounds Medicare's local coverage process, there is little analysis to inform the discussion. To expand knowledge of local coverage policy, this paper traces its origins and evolution. I conclude that the focus on locus, framing the debate in terms of local versus national, obscures fundamental policy issues of access, equity, and quality in Medicare. PMID- 12889762 TI - Health literacy: a policy challenge for advancing high-quality health care. AB - Health literacy, at the intersection of health and education, involves more than reading ability. Studies of health literacy abilities show that many Americans with the greatest health care needs have the least ability to comprehend information required to navigate and function in the U.S. health care system. This paper defines health literacy as an important policy issue and offers strategies for creating a health-literate America. PMID- 12889763 TI - Trends in medication use and functioning before retirement age: are they linked? AB - This paper explores the extent to which changes in medication use during the 1990s are linked to improvements in functioning among Americans before they reach retirement age. Using two cross-sections from a survey of Americans ages 51-61, we examined changes between 1992 and 1998 in the prevalence of functional limitations and medication use associated with five chronic conditions: hypertension, diabetes, lung disease, stroke, and arthritis. We found no evidence linking increases in medication use to improvements in functioning. Instead, increases in educational attainment offset the negative effects of increases in obesity and arthritis over this period. Findings underscore the need for caution in projecting forward improvements in old-age functioning when considering the future of Medicare, Medicaid, and other programs that serve the elderly with disabilities. PMID- 12889764 TI - SCHIP making progress: increased take-up contributes to coverage gains. AB - Previous research based on the Community Tracking Study (CTS) showed that while coverage expansions through the State Children's Health Insurance Program (SCHIP) greatly increased children's eligibility for public or private health insurance coverage, uninsurance rates remained unchanged because of low take-up by eligible children. However, more recent data show that this is changing. Children's uninsurance rates decreased sharply between 1999 and 2001; these changes were greatest in communities where take-up rates have traditionally been the lowest and uninsurance rates the highest. Although uninsurance rates still can be decreased further, state budget pressures threaten the momentum toward higher participation in public programs. PMID- 12889765 TI - Shortcomings in Medicare bonus payments for physicians in underserved areas. AB - This study examines trends in Medicare spending for basic payments and bonus payments for physician services provided to beneficiaries residing in nonmetropolitan counties. For our analysis, we used Medicare Part B physician/supplier claims data for 1992, 1994, 1996, and 1998. Payments under the congressionally mandated bonus payment program acccounted for less than 1 percent of expenditures for physician services in nonmetropolitan, underserved counties. Physician payments increased from 1992 to 1998, while bonus payments increased through 1996 but then declined by 13 percent by 1998. The share of bonus payments to primary care physicians declined throughout the decade, but the share for primary care services increased. PMID- 12889766 TI - Maria. An unexpected meeting between a doctor and a girl with AIDS bears fruit for HIV prevention efforts. PMID- 12889767 TI - Kidneys and the kindness of strangers. PMID- 12889768 TI - Do physicians not offer useful services because of coverage restrictions? AB - Ethically, physicians should discuss all medically appropriate services with patients, but coverage restrictions can make these discussions difficult. In a national survey of physicians, we asked how often physicians elected not to offer their patients useful services because of health plan coverage rules. During the course of a year, 31 percent reported having sometimes not offered their patients useful services because of perceived coverage restrictions. Among these, 35 percent reported doing so more often in the most recent year than they did five years ago. It can be frustrating for doctors to discuss uncovered services with their patients, but open communication is necessary for shared decision making and to improve coverage decisions. PMID- 12889769 TI - Raising the Medicare eligibility age: effects on the young elderly. AB - If Medicare eligibility were delayed to age sixty-seven, as proposed periodically by policymakers, most sixty-five- and sixty-six-year-olds (the "young elderly") would find alternative sources of coverage. However, the loss of Medicare eligibility would leave about 9 percent of this age group uninsured, while another 11 percent would be underinsured because they could only afford limited nongroup policies. The impact would be much greater for certain subgroups, including blacks, Hispanics, and the poor. A buy-in plan that allows young elderly people to purchase Medicare coverage could reduce uninsurance rates, but only if it subsidizes premiums for those with limited incomes. PMID- 12889770 TI - The uninsured and the benefits of medical progress. AB - In a recent Health Affairs article, David Cutler and Mark McClellan found that new medical technology confers positive net benefits for several conditions, including heart attacks, cataracts, and depression. We estimate the extent to which uninsured Americans ages 55-64 use these technologies and compute access gaps for each. Based on Cutler and McClellan's net benefit estimates, we calculate that more than $1.1 billion is lost annually from excess morbidity and mortality among the uninsured population because of lack of access to new technologies for the treatment of these three conditions. PMID- 12889771 TI - Medication costs, adherence, and health outcomes among Medicare beneficiaries. AB - In a two-year period more than two million elderly Medicare beneficiaries did not adhere to drug treatment regimens because of cost. This poor adherence tended to be more common among beneficiaries with no or partial medication coverage and was associated with poorer health and higher rates of hospitalization. The risk for cost-related poor adherence was especially pronounced among lower-income beneficiaries with high out-of-pocket drug spending. We argue that this pattern of cost-related poor medication adherence should inform the design of Medicare prescription drug benefit legislation. PMID- 12889772 TI - Leveraging the nation's anti-bioterrorism investments: foundation efforts to ensure a revitalized public health system. AB - The emerging potential threats of bioterrorism combined with critical existing epidemics facing the United States call for immediate and urgent attention to the U.S. public health system. The foundation world is helping to answer that call and is sounding the alarm that our health defenses must be able to do "double duty" to protect us from the full spectrum of modern health threats. This Special Report presents a selective sample of recent and ongoing grant activities designed to revitalize and modernize the public health infrastructure, which is vital to protecting the nation's health and ensuring its safety. PMID- 12889773 TI - A prescription for change: the need for qualified physician leadership in public health. AB - A key element missing in the federal bioterrorism preparedness plan is qualified physician leadership at the local level. Physicians now lead fewer than one fourth of local health departments. When appointed leaders are not physicians, leadership falls on elected officials or non-medical administrators who become managers of outbreaks. As illustrated in recent case examples, these leaders may find themselves in medical emergencies that they are not qualified to handle. In serious disease outbreaks, unprepared leadership could contribute to unnecessary illness and death. Here I propose strategies to increase qualified physician leadership in state and local public health infrastructures. PMID- 12889774 TI - Drug costs in Canada. PMID- 12889775 TI - Easy ride for drug industry. PMID- 12889776 TI - Medicaid can push for quality. PMID- 12889777 TI - Physician shortages. PMID- 12889778 TI - Bad nursing homes' human toll. PMID- 12889779 TI - Apparent solubility of drugs in partially crystalline systems. AB - Using several griseofulvin samples, representing different solid-state structures, the solubility behavior of drugs in both one-state (totally ordered, semiordered or disordered) and two-state systems was studied. Special attention was directed towards the surface structure of the particles. The partially crystalline samples were obtained by milling the raw material (crystalline standard) or storing the quenched sample (amorphous standard). The solid-state structure of the materials was studied using x-ray diffraction (XRD), differential scanning calorimetry (DSC), isothermal microcalorimetry (IMC), and scanning electron microscopy (SEM). The saturation concentration of the materials was studied in suspensions containing different dispersion concentrations of drug after centrifugation and filtration, using spectrophotometry. In all cases these dispersion concentrations exceeded the solubility of the drug. The solubilities were plotted vs. dispersion concentrations for each sample. Several solubility plateaus were found. The lowest and highest solubility plateaus corresponded to the solubilities of crystalline and amorphous standards. These plateaus were reached at 8 and 44 microg/mL for crystalline and amorphous griseofulvin standards, respectively. An intermediate plateau served as an indication of the existence of a totally semiordered structure. This was reached at 19 microg/mL for griseofulvin. Any deviation from these plateaus was suggested to be indicative of the existence of heterogeneity on the surface structure, which in most cases could be described as a two state system. In such cases, the apparent solubility was a function of dispersion concentration, until at very high dispersion concentrations (4000-20,000 microg/mL) the saturation concentration of the totally disordered (44 microg/mL) or semiordered (19 microg/mL) one-state phase was reached. No reduction in these values was observed during storage for 50 days. It is thus concluded that, in partially crystalline systems, the saturation concentration is an interfacial phenomenon, which depends on the amount, reactivity, and solid-state structure of the exposed solid surfaces in equilibrium with the solution. A simplified solubility model is proposed to qualitatively describe the relationship between established apparent solubilities (saturation concentrations) and different combinations of solid-state structures. PMID- 12889780 TI - Time-dependent mechanical properties of polymeric coatings used in rupturable pulsatile release dosage forms. AB - The mechanical properties of polymer films used in pharmaceutical coatings of pulsatile drug delivery systems were evaluated in the dry and the wet state by a newly developed puncture test, which allowed the time-dependent measurement of the mechanical properties on the same film specimen. Force, puncture strength, energy at break, modulus, and strain were investigated as a function of water exposure time with respect to the type of polymer and the type and concentration of plasticizer and pore former (hydroxypropyl methylcellulose, HPMC). Eudragit RS films were very flexible, had a high strain, and broke upon puncture with only small cracks. In contrast, ethylcellulose films were more brittle with a lower strain and showed complete film rupture. Increased amounts of the hydrophilic pore former, HPMC, resulted in a reduced puncture strength and in an increase in water uptake and weight loss of the films. The puncture strength decreased with increasing plasticizer concentration and was lower with the lipophilic dibutyl sebacate than with the hydrophilic triethyl citrate. PMID- 12889781 TI - Influence of the storage orientation on the aerodynamic particle size of a suspension metered dose inhaler containing propellant HFA-227. AB - Presented in this work are the results of a study designed to investigate the impact of the storage position on the particle size distribution (PSD) of a steroid suspension metered dose inhaler (MDI) containing propellant HFA-227. It was hypothesized that the orientation of MDI samples upon storage could influence the PSD of the emitted dose, since it determines the amount of contact the liquid formulation has with the valve and therefore the quantity of nonvolatile leachable materials from the valve components that may enter the product and potentially impact the aerosol spray dynamics. Samples stored in the valve down orientation (i.e., complete contact of the liquid formulation with the valve) showed a higher level of leachables compared to those samples stored valve up (i.e., minimal contact of the formulation with the valve). The valve down samples were found to produce larger particles in the emitted aerosol spray using both cascade impaction, the preferred method of regulatory submission, as well as laser diffraction. It was postulated that the larger particle size of the inverted samples was attributed to its higher levels of leachables. Based on our findings, it is recommended that in order to set appropriate controls on the product PSD, the storage orientation of the product will need to be considered. PMID- 12889782 TI - Itraconazole formulation studies of the melt-extrusion process with mixture design. AB - Itraconazole is a poorly water soluble compound. One method to increase the aqueous solubility of itraconazole is through formation of a solid dispersion. The purpose of this study is to develop a 40% w/w itraconazole formulation through solid dispersion formation, using hydroxypropyl-beta-cyclodextrin (HP beta-CD) and hydroxypropylmethyl-cellulose (HPMC) as mixture components. The solid dispersion was obtained by melt-extrusion using a twin-screw corotating melt extruder. A D-optimal mixture design was applied for the development of the optimal itraconazole formulation. The itraconazole fraction varied between 20% w/w and 50% w/w in the mixture design and the HPMC and HP-beta-CD fractions varied between 10% w/w and 60% w/w. The itraconazole formulation was optimized by producing clear extrudates, minimizing the torque, and maximizing the glass transition temperature and the apparent itraconazole solubility in 0.1 N HCl. Regression models were developed for the torque, glass transition temperature, and apparent solubility of itraconazole. High itraconazole fraction in the mixture promoted a better melt processing (minimizes torque). High HPMC fraction (>33% w/w) resulted in clear extrudates, indicating a solid dispersion and resulted in high glass transition temperature of the melt. High HP-beta-CD fraction resulted in increased apparent itraconazole solubility in 0.1 N HCl. The optimal itraconazole formulation consisted of 45% w/w HPMC and 15% HP-beta-CD w/w. PMID- 12889783 TI - Average bioequivalence of clarithromycin immediate released tablet formulations in healthy male volunteers. AB - The objective of this study was to assess average bioequivalence of two immediate released tablet formulations of 500-mg clarithromycin tablets in 24 healthy Thai male volunteers. In a randomized, single dose, fasting state, two-period, crossover study design with a 1-week washout period, each subject received a 500 mg clarithromycin tablet. Plasma samples were collected over a 24-hour period after oral administration and were analyzed by using a validated method using high performance liquid chromatography with electrochemical detection. Pharmacokinetic parameters were determined by using noncompartmental analysis. The time to reach the maximal concentration (tmax, h), the peak concentration (Cmax, ng/mL), and the area under the curve (AUC0-infinity, ng x h/mL) of the Reference and Test formulations were 2.0 +/- 0.8 vs. 2.2 +/- 0.9, 2793 +/- 1338 vs. 2642 +/- 1344, and 17912 +/- 7360 vs. 17660 +/- 7992, respectively. Relative bioavailability was 0.99. The 90% confidence interval of Cmax and AUC0-infinity were 82.6-112.1% and 84.7-112.0%. Bioequivalence between the Test and Reference formulation can be concluded. PMID- 12889784 TI - A novel pH- and time-dependent system for colonic drug delivery. AB - A novel pH- and time-dependent delivery system was developed for delivering drugs to the colon. In vitro studies showed that this novel system could release the drug at a predetermined time, which was mainly controlled by the coating layers of the system. The delayed time of the press-coating layer was controlled by its erosion rate, which followed Hixson-Crowell equation. A proper selection of such factors as the viscosity grade of HPMC and tablet hardness, etc., can help reproduce the drug release profile as expected. The transit profiles in two healthy volunteers by gamma scintigraphy demonstrated that the tablets were able to pass through the stomach and small intestine intact and could safely reach the distal end of the small intestine, where the system began to release the drug contained in the core tablet. For both of the volunteers, disintegration of the tablets occurred in the ascending colon, which had highlighted the potential of this system for colonic drug delivery. PMID- 12889785 TI - Biodegradation studies of rosin-based polymers. AB - This study was designed to investigate two rosin-based polymers (R-1 and R-2) for their in vitro and in vivo biodegradation behavior. The in vitro hydrolytic degradation was carried out in buffer solutions of pH 4.4, 7.4, and 10.4 at 37 degrees C. Enzymatic degradation was studied using enzymes lipase, pancreatine, and pectinase. Free films of the two polymers were subcutaneously implanted in rabbits for the in vivo biodegradation. The extent of degradation was determined quantitatively by weight loss and was followed qualitatively by scanning electron microscopy. The extent and the rate of degradation was better in vivo than in vitro. The polymers showed poor enzymatic degradation and a highly pH-dependent hydrolytic degradation. PMID- 12889786 TI - A homogeneity study using NIR spectroscopy: tracking magnesium stearate in Bohle bin-blender. AB - A method was developed for studying mixing of cohesive pharmaceutical mixtures. A combination of accurate sampling and NIR spectroscopic analysis was developed as a suitable method to determine homogenization of magnesium stearate as a function of blending variables. A typical pharmaceutical blend containing a ratio 35:64:1 lactose, avicel, and magnesium stearate was used as a model system. The method accounted for variability of the concentration of magnesium stearate as well as variability of the excipients. Levels of magnesium stearate as low as 0.05% could be resolved by the method, and showed a predicting confidence interval above 98%. PMID- 12889787 TI - Physical characterization and macrophage cell uptake of mannan-coated nanoparticles. AB - Previously, we reported on a cationic nanoparticle-based DNA vaccine delivery system engineered from warm oil-in-water microemulsion precursors. In these present studies, the feasibility of lyophilizing the nanoparticles and their thermal properties were investigated. Also, the binding and uptake of the nanoparticles by a macrophage cell line were studied. The nanoparticles (prior to pDNA coating) were freeze-dried with lactose or sucrose as cryoprotectants. The stability of lyophilized nanoparticles at room temperature was monitored and compared to that of the aqueous nanoparticle suspension. The thermal properties of the nanoparticles were investigated using differential scanning calorimetry (DSC). The nanoparticles, coated or uncoated with mannan as a ligand, were incubated with a mannose receptor positive (MR+) mouse macrophage cell line (J774E), at either 4 degrees C or 37 degrees C to study the binding and uptake of the nanoparticles by the cells. It was found that lactose or sucrose (1-5%, w/v) was required for successful lyophilization of the nanoparticles. After 4 months of storage, the size of lyophilized nanoparticles did not significantly increase while those in aqueous suspension grew by over 900%. Unlike its individual components, emulsifying wax (m.p., approximately 55 degrees C) and hexadecyltrimethyl ammonium bromide, the nanoparticles showed a melting point of approximately 90 degrees C. Moreover, the DSC profile of the nanoparticles was different from that of the physical mixture of emulsifying wax and CTAB. After 1 hour incubation at 37 degrees C, the uptake of mannan-coated nanoparticles was 50% higher than that of the uncoated nanoparticles. At 4 degrees C and after one hour, the binding of the mannan-coated nanoparticles by J774E was over 2-fold higher than that of the uncoated nanoparticles. This increase in J774E binding could be abolished by preincubating the cells with free mannan, suggesting that the binding and uptake were receptor-mediated. In conclusion, the nanoparticles were lyophilizable, and lyophilization was shown to enhance the stability of the nanoparticles. DSC provided evidence that the nanoparticles were not a physical mixture of their individual components. Finally, cell binding and uptake studies demonstrated that the nanoparticles have potential application for cell-specific targeting. PMID- 12889788 TI - Polymer-stabilized emulsion systems: structural characteristics and physical stability evaluation. AB - In the course of investigation of the effects of polymeric emulsifier (Simulgel A) on the structural characteristics and physical stability (during storage time) of semisolid o/w emulsion systems with different emulsifier concentrations, dispersed droplets of an internal oil phase (4.5-11.5 microm size) were discovered. The degree of dispersity was increased along with increasing polymeric emulsifier concentration (from 1 to 5% m/m), which was shown by microscopic analysis. Experiments were also carried out using pH, electrical conductivity, and rheological (rotational and oscillatory) measurements. Emulsion samples expressed a mild acid value acceptable for topical preparations during a 6-month period. Electrical conductivity of fresh samples ranged from 0.423 to 2.009 mS/cm and after a 6-month period this slightly increased to 0.663-2.890 mS/cm. All emulsion samples revealed plastic flow behavior in fresh state, with changes in flow pattern during storage time. Viscoelastic properties defined by the oscillatory rheometry results were in good correlation with parameters of rotational rheometry and degree of internal phase dispersity. High yield stress values (flow tests) and favorable low values of damping factor (oscillatory studies) both indicated good internal structuring and long-term stability of sample containing 3% m/m of polymeric emulsifier. PMID- 12889789 TI - Chitosan-alginate multilayer beads for gastric passage and controlled intestinal release of protein. AB - Chitosan-alginate beads loaded with a model protein, bovine serum albumin (BSA) were investigated to explore the temporary protection of protein against acidic and enzymatic degradation during gastric passage. Optimum conditions were established for preparation of homogenous, spherical, and smooth chitosan alginate beads loaded with BSA. Multilayer beads were prepared by additional treatment with either chitosan or alginate or both. The presence of chitosan in the coagulation bath during bead preparation resulted in increased entrapment of BSA. During incubation in simulated gastric fluid (SGF pH 1.2), the beads showed swelling and started to float but did not show any sign of erosion. Inclusion of pepsin in the gastric fluid did not show a further effect on the properties of the beads. Release studies were done in simulated gastric fluid (SGF pH 1.2) and subsequently in simulated intestinal fluid (SIF pH 7.5) to mimic the physiological gastrointestinal conditions. After transfer to intestinal fluid, the beads were found to erode, burst, and release the protein. Microscopic and macroscopic observations confirmed that the release of protein was brought about by the burst of beads. Chitosan-reinforced calcium-alginate beads showed delay in the release of BSA. The multilayer beads disintegrated very slowly. The enzymes pepsin and pancreatin did not change the characteristics of BSA-loaded chitosan alginate beads. Single layer chitosan-alginate beads released 80-90% of the model protein within 12h while multilayer beads released only 40-50% in the same period of time. The release from chitosan-alginate beads and multilayer beads in SIF was further delayed without prior incubation in SGF. It is concluded that alginate beads reinforced with chitosan offer an excellent perspective for controlled gastrointestinal passage of protein drugs. PMID- 12889790 TI - Anthocyanic vacuolar inclusions (AVIs) selectively bind acylated anthocyanins in Vitis vinifera L. (grapevine) suspension culture. AB - Anthocyanic vacuolar inclusions (AVIs) appear as dark red-to-purple spheres of various sizes in vacuoles of grapevine (Vitis vinifera L.) cell suspension culture due to their interaction with anthocyanins. AVIs were purified and the bound anthocyanins extracted and analysed by HPLC from two lines of V. vinifera isolated from the same callus accumulating anthocyanin in the dark, yet varying in their anthocyanin profiles and accumulation. An intermediate-pigmented line (FU-1) with a 1.3:1 ratio of acylated:non-acylated anthocyanins, a colour value of 0.84 units and cyanidin and peonidin as the dominant species was compared with a high-pigmented line (FU-2) with a 1.2:1 ratio of acylated:non-acylated anthocyanins, a colour value of 3.72 units and malvidin predominating. The profile of AVI-bound anthocyanins showed an increase in acylated anthocyanins in both lines of approx. 28-29%, with no apparent preference for anthocyanin species. This resulted in a ratio of acylated:non-acylated anthocyanins of 6.2:1 for FU-1 and 4.9:1 for FU-2. The reasons for the selectivity of the AVIs for acylated (specifically p-coumaroylated) species compared with the whole cell profile are discussed. PMID- 12889791 TI - Screening of yeasts for cell-free production of (R)-phenylacetylcarbinol. AB - 105 yeast strains from 10 genera and 40 species were evaluated for cell-free production of (R)-phenylacetylcarbinol (PAC), the chiral precursor in the manufacture of the pharmaceuticals ephedrine and pseudoephedrine. Carboligase activity of pyruvate decarboxylase (PDC), forming PAC from benzaldehyde and pyruvate, was found in extracts of 98 strains. PAC was not formed from benzaldehyde and acetaldehyde, an activity of bacterial PDCs from Zymomonas mobilis and Zymobacter palmae. Two interesting groups of candidates were identified in the yeast screening: carboligase activities of Schizosaccharomyces pombe PDCs were very low but showed best resistance to pre-incubation with acetaldehyde and benzaldehyde; and highest carboligase activities combined with medium resistance were found in strains of Candida utilis, C. tropicalis and C. albicans. PMID- 12889792 TI - Increased pyruvate efficiency in enzymatic production of (R) phenylacetylcarbinol. AB - Loss of substrate, pyruvate, a limitation for enzymatic batch production of (R) phenylacetylcarbinol (PAC), resulted from two phenomena: temperature dependent non-enzymatic concentration decrease due to the cofactor Mg2+ and formation of by products, acetaldehyde and acetoin, by pyruvate decarboxylase (PDC). In the absence of enzyme, pyruvate stabilization was achieved by lowering the Mg2+ concentration from 20 to 0.5 mM. With 0.5 mM Mg2+ Rhizopusjavanicus and Candida utilis PDC produced similar levels of PAC (49 and 51 g 1(-1), respectively) in 21 h at 6 degrees C; however C. utilis PDC formed less by-product from pyruvate and was more stable during biotransformation. The process enhancements regarding Mg2+ concentration and source of PDC resulted in an increase of molar yield (PAC/consumed pyruvate) from 59% (R. javanicus PDC, 20 mM Mg2+) to 74% (R. javanicus PDC, 0.5 mM Mg2+) to 89% (C. utilis PDC, 0.5 mM Mg2+). PMID- 12889793 TI - Estimation of Chinese hamster ovary cell density in packed-bed bioreactor by lactate production rate. AB - A method is described for estimating recombinant Chinese hamster ovary (rCHO) cell density in a packed-bed bioreactor by lactate production rate. The lactate production rate, which depended on both the cell numbers and cell growth rate, was modeled by segregating the cell population into two parts: one growing at a maximum specific growth rate and another non-growing. The individual cell in each part had the same lactate production rate. The established rate equation of lactate production matched the experimental data reasonably well and could be used to estimate the cell growth in the batch culture with microcarriers. Furthermore, in the perfusion culture of rCHO cells in a packed-bed bioreactor, the final cell density, 1.3 x 10(10) cells l(-1), estimated by lactate production rate, was comparable to the direct sample counting of 1.2 x 10(10) cells l(-1), showing that lactate production rate method would be useful in tracing the cell growth in packed-bed bioreactors. PMID- 12889794 TI - A bioassay for methylated galacturonan on pulp-fiber surfaces. AB - A colorimetric bioassay has been developed enabling separation of wood pulp qualities based on the quantity of methyl-esterified galacturonan exposed on fiber surfaces. Thermomechanical pulp (TMP) and chemi-TMP (CTMP) had, respectively, 3.1 and 0.7 microg galacturonan on the surface per mg wood pulp. The presence of galacturonan on the surface of TMP and CTMP was further confirmed by immunogold localization. PMID- 12889795 TI - Quantification and characterization of insoluble chitinous materials in viscous chitosan solutions. AB - Insoluble chitinous materials in highly viscous chitosan solutions can be quantified using the viscosity-lowering action of transglucosidase (EC 2.4.1.24). In chitosan, commonly produced by high temperature deacetylation (90 degrees C), between 70-90% of insoluble chitinous materials were recovered by this enzymatic method whereas only 25% recovery was obtained by the nitrous acid method. The insoluble material recovered after enzyme treatment had a higher degree of deacetylation and a lower degree of crystallization than that after nitrous acid treatment. The results are explained by difference in penetration by enzyme and nitrous acid into the insoluble particle. PMID- 12889796 TI - Immobilization of pectinmethylesterase from acerola (Malpighia glabra L.) in porous silica. AB - The total and partially purified enzyme pectinmethylesterase from acerola fruit was covalently immobilized on porous silica particles. These efficiency values were 114% for the total PME and 351% for the partially purified PME. In both forms the immobilization resulted in compounds with high thermal stability. PMID- 12889797 TI - Enhanced shear protection and increased production of an anti-tumor polysaccharide by Agaricus blazei in xanthan-supplemented cultures. AB - Xanthan supplementation provided shear protection and stimulated polysaccharide production by Agaricus blazei. In xanthan-free cultures, the optimal cell yield, 0.63 g biomass g(-1) glucose, and product yield, 0.19 g polysaccharide g(-1) glucose, were, respectively, when the critical impeller tip speed was 50.3 cm s( 1) and 100.5 cm s(-1). Furthermore, the critical impeller tip speed of cell yield shifted from 50.3 cm s(-1) to 100.5 cm s(-1) with the supplementation of 1 g xanthan l(-1). Maximum specific product yield, namely 0.74 g polysaccharide g(-1) biomass, was achieved with inlet air supply of 3% O2 and impeller tip speed of 100.5 cm s(-1). PMID- 12889798 TI - Application of statistically-based experimental designs for the optimization of nisin production from whey. AB - Statistically-based experimental designs were applied for the optimization of nisin production by Lactococcus lactis in a whey-based medium. Yeast extract, KH2PO4, and MgSO4 were identified to have significant effects on nisin biosynthesis by a Plackett-Burman design. These three significant factors were subsequently optimized using central composite design, and the optimal conditions were determined to be 12.067 g l(-1) for yeast extract, 0.569 g l(-1) for KH2PO4, and 0.572 g l(-1) for MgSO4. The validity of the optimal conditions was verified by a separate experiment. PMID- 12889799 TI - A peptide antibody for rapid screening of Streptomyces species producing phospholipase D. AB - By examining the conserved regions in the protein sequences of eight different Streptomyces phospholipase Ds (PLD) reported so far and the X-ray crystallographic structure of a Streptomyces PLD, we designed a peptide sequence, DPANRGAVGSGGYSQIKSL, for the screening of microorganisms producing PLD. In the enzyme-linked immunosorbent assay using a mouse antibody raised against the designed peptide, we recovered seven producing strains out of 128 soil isolates. PMID- 12889800 TI - Application of a two-stage temperature control strategy for enhanced glutathione production in the batch fermentation by Candida utilis. AB - In batch culture for glutathione production with Candida utilis, a higher temperature (30 degrees C) was required to hasten cell growth while a lower temperature (26 degrees C) was needed to increase the production of glutathione. A two-stage temperature control strategy was used to enhance both the yield and the productivity of glutathione. As a result, glutathione production was increased by 5% and 23% of that at 26 degrees C and 30 degrees C, respectively, and the intracellular glutathione content reached 2.5% (w/w). PMID- 12889801 TI - A screening method for detecting iron reducing wood-rot fungi. AB - A plate assay using the Fe(II) selective dye, ferrozine, for detecting wood-rot fungi with Fe(III) reductive abilities, was developed. The assay is fast, simple and, in most cases, more sensitive than the corresponding liquid medium test. The brown rot fungi, Gloeophyllum trabeum and Laetiporeus sulphureus, displayed higher iron reductive capabilities than white rot fungi, Trametes versicolor, Ganoderma australe and Ceriporiopsis subvermispora. PMID- 12889802 TI - Laser-assisted microinjection into targeted animal cells. AB - A pulsed (17 nanoseconds) Nd:YAG laser (1064 nm) was used to inject impermeable dyes (propidium iodide andiodide and merocyanine 540) and a plasmid (pEGFP-N1) encoding green fluorescent protein (GFP) into human breast adenocarcinoma cells (MCF-7). The cell membrane integrity and viability were fully preserved in this laser-assisted transfer. PMID- 12889803 TI - Assembly of linear functional expression elements with DNA fragments digested with asymmetric restriction endonucleases. AB - A three-step method is described to assemble linear functional expression elements for rapid gene expression. In step one, a eukaryotic promoter, an open reading frame and a transcription terminator are PCR-amplified individually with primers containing recognition sites of asymmetric restriction endonucleases. In step two, the three DNA fragments, digested with corresponding restriction endonucleases, are directionally joined by T4 DNA ligase to assemble linear functional expression elements. In step three, the resultant linear functional expression elements are amplified with element-specific primers by a second round PCR, followed by transfection into Chinese hamster ovary cells for gene expression. PMID- 12889804 TI - Stability of tumor necrosis factor-alpha during ion exchange chromatography. AB - Denaturation of target protein by various separation and purification steps contributes significant part to the total product loss in bioseparation. The conformational change and accompanying loss of activity of tumor necrosis factor alpha during ion exchange chromatography was reversible and was decreased by adding polyethylene glycol 200 at 2 to 5% (v/v) to the eluting solution. PMID- 12889805 TI - Functional identification of rub52 gene involved in the biosynthesis of rubradirin. AB - An open reading frame, rub52, has been identified as a gene encoding thymidine diphospho-glucose 2,3-dehydratase by sequence analysis of the rubradirin biosynthetic gene cluster of Streptomyces achromogenes var. nibradiris NRRL3061. The gene codes for a protein consisting of 458 amino acids with calculated molecular mass of 50862 Da. The gene was amplified and heterologously expressed in Escherichia coli as a soluble His-tagged fusion protein. C-2 deoxygenation functionality of thymidine diphospho-4-keto-6-deoxyglucose was assigned to the rub52 gene product from in vitro enzyme assay. PMID- 12889806 TI - Resonant Raman quantification of zeaxanthin production from Flavobacterium multivorum. AB - Resonant Raman scattering was used as a novel, rapid, non-destructive optical technique to measure zeaxanthin levels in Flavobacterium multivorum ATCC 55238. Culture broth, after bacterial growth for 40 h, exhibited characteristic resonance Raman vibrational modes at 1159 cm(-1) (C-C stretch) and 1525 cm(-1) (C=C stretch) upon excitation at 488 nm. A striking correlation was observed between the carotenoid level as estimated by HPLC and by resonance Raman spectroscopy. PMID- 12889807 TI - Functional expression of arginine kinase improves recovery from pH stress of Escherichia coli. AB - Acid stress in Escherichia coli involves a complex resource- and energy-consuming response mechanism. By overexpression of arginine kinase from Limulus polyphemus in E. coli, we improved the recovery from a transient pH stress. While wild type E. coli resumed growth after a transient pH reduction to pH 3 for 1 h with a rate that was 25% lower than before the stress, the arginine kinase expressing strain continued to grow as rapidly as before. This effect is presumably caused by the physiological function of arginine kinase as a short term energy buffer in the form of phosphoarginine, but a pH-buffering effect cannot be excluded. PMID- 12889808 TI - Functional expression and production of human H-ferritin in Pichia pastoris. AB - Human heavy chain ferritin (H-ferritin) was cloned from human heart cDNA library and expressed in Pichia pastoris. The H-ferritin transformant was cultivated by fed-batch and the cell mass reached about 52 g cell dry wt l(-1) after 150 h. In atomic absorption spectrometry analysis, intracellular content of iron in H ferritin transformant was measured to 3038 +/- 72 microg g(-1) which was 9.6-fold more than that of control strain. PMID- 12889809 TI - Nucleofection as an efficient nonviral transfection method for human monocytic cells. AB - Despite some progress in the field of gene transfer into hard-to-transfect cells, so far an efficient nonviral method for monocytes has not been available. A comparison of plasmid DNA with capped and polyadenylated mRNA for enhanced green fluorescent protein gene delivery into the commonly used monocytic cell lines U937 and THP-1 suggested that limited DNA trafficking may be the underlying cause of poor transfection results. As Nucleofector technology delivers DNA (or mRNA) straight into the nucleus, we obtained nucleofection efficiencies of up to 80% without significant cell toxicity. Moreover, as the DNA quickly reaches the nucleus, nucleofected cells were ready for analysis after only 2-6 h. The technique is suitable not only for monocytes but also for other hard-to-transfect cells. PMID- 12889810 TI - Multiple displacement amplification prior to single nucleotide polymorphism genotyping in epidemiologic studies. AB - We assessed the whole genome amplification strategy, known as multiple displacement amplification (MDA), for use with the TaqMan genotyping platform for DNA samples derived from two case-control studies nested in the Nurses' Health Study and the Physicians' Health Study. Our objectives were to (1) quantify DNA yield from samples of varying starting concentrations and (2) assess whether MDA products give an accurate representation of the original genomic sequence. Multiple displacement amplification yielded a mean 23000-fold increase in DNA quantity and genotyping results demonstrate 99.95% accuracy across six SNPs from four genes for 352 samples included in this study. These results suggest that MDA will provide a sufficiently robust amplification of limiting samples of genomic DNA that can be used for SNP genotyping in large case-control studies of complex diseases. PMID- 12889811 TI - Fusion protein of the hyaluronan binding domain from human TSG-6 with luciferase for assay of hyaluronan. AB - The gene expression plasmid, pET-Lmluc, for the fusion protein of the hyaluronan binding domain from human TSG-6 [product of tumor necrosis factor (TNF) stimulated gene-6] and luciferase from Renilla reniformis was constructed. The fused gene was expressed in Escherichia coli and the resulted insoluble Lm-luc fusion protein was purified and refolded to recover both the hyaluronan binding capability and the luciferase activity. Hyaluronan as low as 1 ng ml(-1) was detected by using the indirect enzymatic immunological assay with the refolded Lm luc fusion protein. PMID- 12889812 TI - A single residual replacement improves the folding and stability of recombinant cassava hydroxynitrile lyase in E. coil. AB - Substitution of Ser113 for Gly113 in the cap domain of hydroxynitrile lyase from Manihot esculenta (MeHNL) was performed by site-directed mutagenesis to improve its self-generated folding and stability under denaturation conditions. The yield of the recombinant mutant HNL1 (mut-HNL1), which had higher specific activity than the wild type HNL0 (wt-HNL0), was increased by 2 to 3-fold. Thermostability of MeHNL was also enhanced, probably due to an increase in content of the beta strand secondary structure according to CD analysis. Our data in this report suggest that Ser113 significantly contributes to the in vivo folding and stability of MeHNL and demonstrates an economic advantage of mut-HNL1 over the wt HNL0. PMID- 12889813 TI - Purification and properties of a heat-stable inulin fructotransferase from Arthrobacter ureafaciens. AB - An inulin fructotransferase producing difructose dianhydride I (EC 2.4.1.200) was purified from Arthrobacter ureafaciens A51-1. It had maximum activity at pH 5.5 and 45 degrees C, and was stable up to 80 degrees C. This is the highest thermal stability for this enzyme reported to date. The molecular mass was estimated to be 38000 by SDS-PAGE, and 61000 by gel filtration. It was therefore estimated to be a dimer. PMID- 12889814 TI - Production of alkaloids by in vitro culture of Erythrina americana Miller. AB - The production of erythroidines and other alkaloids was studied in cotyledons, callus and cell suspension cultures of Erythrina americana Miller. The cell suspension cultures, grown in Murashige & Skoog medium with naphthaleneacetic acid (3 mg l(-1)) and kinetin (2 mg l(-1)), produced 89 and 17 microg alpha- and beta-erythroidines respectively per g dry wt. PMID- 12889815 TI - Multitasking with molecular dynamics Typhoon: quantifying nucleic acids and autoradiographs. AB - With increased sensitivity and specificity, fluorescent assays are rapidly becoming the method of choice for nucleic acid quantification. The utility of the Typhoon scanner has now been extended to accurately measure low levels of DNA and RNA (5 ng ml(-1)) with PicoGreen and RiboGreen dyes. In addition, with a few simple modifications, autoradiographic film images can be scanned and quantified with the Typhoon series of scanners. PMID- 12889816 TI - Catalytic analysis of a recombinant D-hydantoinase from Agrobacterium tumefaciens. AB - The D-hydantoinase gene of a wild strain of Agrobacterium tumefaciens BQL9 had 99.78% nucleotide sequence identity with other available Agrobacterium genes. The resulting amino acid sequence showed two important substitutions affecting two alpha-helixes in the secondary structure of the protein. The union of Mn2+ to the protein was essential for activating the enzyme and was independent of the temperature. D-Hydantoinase only was inactivated in the presence of 70 mM EDTA and at over 40 degrees C. The enzyme showed both hydantoinase and pyrimidinase activities, but only with the D-enantiomers of the substrates. Activity was greater for substrates with apolar groups in the number 5 carbon atom of the hydantoin. The native structure of the N-terminal end of this D-hydantoinase proved to be indispensable to its enzymatic activity. PMID- 12889817 TI - Enhancement of the catalytic activity of an artificial phosphotriesterase using a molecular imprinting technique. AB - An artificial phosphotriesterase (PTE) was constructed by co-polymerization of 4(5)-vinylimidazole-Zn(2+)-methacrylic acid cluster with a divinylbenzene polymer. Compared with the spontaneous hydrolysis, the resulting polymer catalyst caused 105-fold rate acceleration towards the hydrolysis of diethyl p-nitrophenyl phosphate (Paraoxon). The catalytic activity of the polymer catalyst could be enhanced for 30% using molecular imprinting technique and the molecularly imprinted catalyst (MIC) showed a turnover rate of 7.4 x 10(-2) s(-1) towards the hydrolysis of Paraoxon. The MIC also hydrolyzed thiophosphates and phosphorothiolate triester pesticides. Construction of an amperometric sensor employing the MIC as catalyst achieved a detection limit of 0.1 mM Paraoxon. PMID- 12889818 TI - Enzymatic resolution of homoallyllic alcohols using various Rhizopus species. AB - The asymmetric resolution of various 1-aryl-3-buten-1-ols via microbial hydrolysis of the corresponding acetates has been investigated using different Rhizopus species. The chosen species, R. arrhizus (wild type), efficiently hydrolyzed 1-phenyl- and 1-para-substituted phenyl-3-buten-1-ol acetates, producing the enantiomerically pure (R)-alcohols with 53-65% yields. Although the antipode acetates were obtained with 9-52% enantiomeric excess, the (S)-alcohols were amenable in >99% enantiomeric excess via a R. arrhizus mediated asymmetric reduction of the corresponding ketones. PMID- 12889819 TI - Foot-and-mouth disease virus VP1 protein fused with cholera toxin B subunit expressed in Chlamydomonas reinhardtii chloroplast. AB - A Chlamydomonas reinhardtii chloroplast expression vector, pACTBVP1, containing the fusion of the foot and mouth disease virus (FMDV) VP1 gene and the cholera toxin B subunit (CTB) gene was constructed and transfered to the chloroplast genome of C. reinhardtii by the biolistic method. The transformants were identified by PCR, Southern blot, Western blot and ELISA assays after selection on resistant medium and incubation in the dark. The CTBVP1 fusion protein was expressed in C. reinhardtii chloroplast and accounted for up to 3% of the total soluble protein. The fusion protein also retained both GM1-ganglioside binding affinity and antigenicity of the FMDV VP1 and CTB proteins. These experimental results support the possibility of using transgenic chloroplasts of green alga as a mucosal vaccine source. PMID- 12889820 TI - L-Lactic acid production from raw cassava starch in a circulating loop bioreactor with cells immobilized in loofa (Luffa cylindrica). AB - L-Lactic acid was produced from raw cassava starch, by simultaneous enzyme production, starch saccharification and fermentation in a circulating loop bioreactor with Aspergillus awamori and Lactococcus lactis spp. lactis immobilized in loofa sponge. A. awamori was immobilized directly in cylindrical loofa sponge while the L. lactis was immobilized in a loofa sponge alginate gel cube. In the loofa sponge alginate gel cube, the sponge serves as skeletal support for the gel with the cells. The alginate gel formed a hard outer layer covering the soft porous gel inside. By controlling the rate and frequency of broth circulation between the riser and downcomer columns, the riser could be maintained under aerobic condition while the downcomer was under anaerobic condition. Repeated fed-batch L-lactic acid production was performed for more than 400 h and the average lactic acid yield and productivity from raw cassava starch were 0.76 g lactic acid g(-1) starch and 1.6 g lactic acid l(-1) h(-1), respectively. PMID- 12889821 TI - Partial purification and biochemical characterization of alkaline 5' phosphodiesterase from barley malt sprouts. AB - An alkaline 5'-phosphodiesterase (5'-PDE) from barley (Hordeum distichum) malt sprouts was partially purified by thermal treatment and acetone precipitation to diminish phosphomonoesterase (PME) activity. 5'-PDE was purified 40-fold to a specific activity of 30 U mg(-1) protein with a final yield of about 32%. With synthetic substrate, the enzyme had an optimum pH of 8.9, maximum activity at 70 degrees C over 10 min, and a Km of 0.26 mM. The partially purified enzyme was activated by 10 mM Mg2+ up to 168% of the original activity, while Zn2+, Mn2+ and Cu2+ ions, chelating agent (EDTA) and NaN3 (1-10 mM), and 5'-ribonucleotides (1-5 mM) were inhibitory. Final enzyme preparation was stable over 8 d at 4 degrees C), at 70 degrees C for up to 120 min and without loss of activity over 90 d at 18 degrees C. PMID- 12889822 TI - Purification, characterization and crystallization of a group of earthworm fibrinolytic enzymes from Eisenia fetida. AB - Seven fibrinolytic enzymes were purified from the earthworm Eisenia fetida. The molecular weights of the enzymes were 24663, 29516, 29690, 24201, 24170, 23028 and 29595, and the respective isoelectric points were 3.46, 3.5, 3.5, 3.68, 3.62, 3.94 and 3.46. All the proteases showed different fibrinolytic activity on fibrin plates. Studies on substrate specificity and inhibition indicated that they belonged to different types of serine proteases. N-Terminal sequencing indicated their high homology to those from the earthworm Lumbricus rubellus. All the enzymes have been crystallized. PMID- 12889823 TI - Metal chelate affinity precipitation of RNA and purification of plasmid DNA. AB - The affinity of metal chelates for amino acids, such as histidine, is widely used in purifying proteins, most notably through six-histidine 'tails'. We have found that metal affinity interactions can also be applied to separation of single stranded nucleic acids through interactions involving exposed purines. Here we describe a metal affinity precipitation method to resolve RNA from linear and plasmid DNA. A copper-charged copolymer of N-isopropyl acrylamide (NIPAM) and vinyl imidazole (VI) is used to purify plasmid from an alkaline lysate of E. coli. The NIPAM units confer reversible solubility on the copolymer while the imidazole chelates metal ions in a manner accessible to interaction with soluble ligands. RNA was separated from the plasmid by precipitation along with the polymer in the presence of 800 mM NaCl. Bound RNA could be recovered by elution with imidazole and separated from copolymer by a second precipitation step. RNA binding showed a strong dependence on temperature and on the type of buffer used. PMID- 12889824 TI - Formation of enterovirus-like particle aggregates by recombinant baculoviruses co expressing P1 and 3CD in insect cells. AB - The assembly of enterovirus requires the cleavage of P1 polyprotein by protease 3CD into individual structural proteins. Two recombinant baculoviruses were constructed to encode P1 and 3CD of enterovirus 71 (EV71), respectively. The expressed 3CD successfully cleaved P1 in vitro and in vivo. Also, the co infection in insect cells resulted in crystalline virus-like particle structures morphologically resembling the authentic EV71 aggregates, which are reported for the first time. PMID- 12889825 TI - Biodegradation of nonylphenol in a continuous packed-bed bioreactor. AB - A packed bed bioreactor, with 170 ml glass bead carriers and 130 ml medium, was tested for the removal of the endocrine disrupter, nonylphenol, with a Sphingomonas sp. The bioreactor was first continuously fed with medium saturated with nonylphenol in an attempt to simulate groundwater pollution. At best, nonylphenol was degraded by 99.5% at a feeding rate of 69 ml h(-1) and a removal rate of 4.3 mg nonylphenol day(-1), resulting in a 7.5-fold decrease in effluent toxicity according to the Microtox. The bioreactor was then fed with soil leachates at 69 ml h(-1) from artificially contaminated soil (1 g nonylphenol kg( 1) soil) and a real contaminated soil (0.19 g nonylphenol kg(-1) soil). Nonylphenol was always completely removed from the leachates of the two soils. It was removed by 99% from the artificial soil but only 62% from real contaminated soil after 18 and 20 d of treatment, respectively, showing limitation due to nonylphenol adsorption. PMID- 12889826 TI - Bioconversion of linoleic acid into conjugated linoleic acid during fermentation and by washed cells of Lactobacillus reuteri. AB - Conjugated linoleic acid (CLA) was produced at 300 mg l(-1) after 24 h culture of Lactobacillus reuteri in de Man-Rogosa-Sharpe medium containing 0.9 g linoleic acid (LA) l(-1) and 1.67% (v/v) Tween 80. CLA was mainly located in the extracellular space of the cells. Washed cells previously grown on LA were less active than unadapted washed cells in converting LA into CLA. Most of the CLA transformed by washed L. reuteri cells was located in cells or associated with cells. CLA production by washed L. reuteri cells was most efficient in conversion with 0.45 g LA l(-1) at pH 9.5 and 37 degrees C for 1 h. PMID- 12889827 TI - Transient reporter gene (GUS) expression in creeping bentgrass (Agrostis palustris) is affected by in vivo nucleolytic activity. AB - Leaf and callus tissues of a creeping bentgrass cultivar (Penn A4) had high nuclease activities that degraded exogenously added plasmid DNA. When callus tissue was incubated for 24 h with heparin, spermidine, aurintricarboxylic acid or polyethylene glycol, only heparin and spermidine were effective as in vitro nuclease inhibitors, protecting exogenously added plasmid DNA from degradation. When beta-glucuronidase (GUS) reporter gene activity was evaluated in heparin treated (0.6%), 14-month old callus following microprojectile bombardment, GUS activity increased 1000-fold compared to equivalent aged untreated Penn A4 callus. Similar enhancement from heparin pretreatment (0.6% or 1.2%) was not observed in 6-month old callus. This is likely due to much higher activities of nuclease in the younger callus. PMID- 12889828 TI - Kinetics of beta-glucosidase production by Saccharomyces cerevisiae recombinants harboring heterologous bgl genes. AB - The maximum productivity of beta-glucosidase by Saccharomyces cerevisiae recombinants under the control of GALI promoter was 100 IU l(-1) h(-1). The highest productivity of beta-glucosidase by a S. cerevisiae recombinant was 16 fold more than that supported by Cellulomonas biazotea. The recombinants also co produced ethanol from cellobiose: maximum product yield and productivity were 0.5 and 1.1 g ethanol g(-1) cellobiose and g ethanol l(-1) h(-1), respectively. PMID- 12889829 TI - Adsorption of Ni2+ on the surface of molecularly imprinted adsorbent from Penicillium chysogenum mycelium. AB - The adsorption capacity for Ni2+ on to the surface molecular imprinting adsorbent on Penicillium chysogenum mycelium (the surface-imprinted adsorbent) was 40-45 mg g(-1) (using 200 mg Ni2+ l(-1)), two times of the mycelium adsorbent. The surface imprinted adsorbent had good stability at pH 2 to approximately 8. The optimal concentration of EDTA for desorption was 0.1 to 0.5 g l(-1). The surface imprinted adsorbent could be reused 15 times without losing its uptake. PMID- 12889830 TI - Oxalic acid production from lipids by a mutant of Aspergillus niger at different pH. AB - Crude rapeseed oil and post-refining fatty acids were used as substrates for oxalic acid production by a mutant of Aspergillus niger. Both the final concentration and the yield of the product were highest at pH 4 to 5. With a medium containing 50 g lipids l(-1), production reached a maximum of 68 g oxalic acid l(-1) after 7 d. A high yield of the product (up to 1.4 g oxalic acid g(-1) lipids consumed) was achieved with oil and fatty acids combined. PMID- 12889831 TI - Evidence for stable transformation of Pseudomonas aeruginosa with a pUC-based plasmid. AB - Pseudomonas aeruginosa was transformed with pUC8:16, a pUC-based plasmid bearing the gene (vgb) encoding Vitreoscilla (bacterial) hemoglobin (VHb). Transformation was initially indicated by an increase in ampicillin resistance from 1500 to 2500 mg l(-1). Presence of the plasmid in P. aeruginosa was confirmed by amplification of a portion of vgb from and detection of VHb in the transformant but not the untransformed host. Southern blot analysis further indicated that pUC8:16 existed as an autonomous plasmid rather than integrated into the chromosome of the P. aeruginosa transformant. PMID- 12889832 TI - Production of tagatose by a recombinant thermostable L-arabinose isomerase from Thermus sp. IM6501. AB - A gene (thaI) corresponding to L-arabinose isomerase from Thermus strain IM6501 was cloned by PCR. It comprised 1488 nucleotides and encoded a polypeptide of 496 residues with a predicted molecular weight of 56019 Da. The deduced amino acid sequence had 96.8% identity with the L-arabinose isomerase of Geobacillus stearothermophilus. Recombinant ThaI with N-terminal hexa-tistidine tags was over expressed in Escherichia coli and purified by affinity chromatography using Ni NTA resin. The purified ThaI was thermostable with maximal activity at 60 degrees C at pH 8 for 30 min of reaction. Zn2+ and Ni2+ inactivated the catalytic activity of ThaI, 5 mM Mn2+ enhanced the bioconversion yield by 90%. The bioconversion yield of 54% from D-galactose to D-tagatose was obtained by recombinant ThaI at 60 degrees C over 3 d. PMID- 12889833 TI - Regioselective synthesis of kojic acid esters by Bacillus subtilis protease. AB - The lipophilicity of kojic acid [5-hydroxy-2-(hydroxymethyl)-4H-pyran-4-one] was improved by esterifying kojic acid with either divinyl adipate, vinyl hexanoate, vinyl octanoate or vinyl decanoate using protease from Bacillus subtilis for 7 d. 1H-NMR and 13C-NMR showed that the primary hydroxyl group at the C-7 position of kojic acid was regioselectively esterified to afford 7-O-vinyl adipoyl kojic acid, 7-O-hexanoyl kojic acid, 7-O-octanoyl kojic acid and 7-O-decanoyl kojic acid (13-27% yield). The kojic acid esters had radical scavenging activities, inhibited tyrosinase activity and was biodegradable. PMID- 12889834 TI - Simultaneous and rapid monitoring of biomass and biopolymer production by Sphingomonas paucimobilis using Fourier transform-near infrared spectroscopy. AB - The application of Fourier Transform near infrared spectroscopy (FT-NIRS) to near real-time monitoring of polysaccharide and biomass concentration was investigated using a gellan-producing strain of Sphingomonas paucimobilis grown in a stirred tank reactor. Successful models for both biomass and gellan were constructed despite the physichochemical complexity of the viscous process fluid. Modelling of biomass proved more challenging than for gellan, partly because of the low range of biomass concentration but a model with a good correlation coefficient (0.94) was formulated based on second derivative spectra. The gellan model was highly satisfactory, with an excellent correlation coefficient (0.98), again based on second derivative spectra. No sample pre-treatment was required and all spectral scanning was carried out on whole broth. Additionally, both models should be robust in practice since both were formulated using low numbers of factors. Thus, the near real time simultaneous monitoring of gellan and biomass in this highly complex matrix using FT-NIRS potentially opens the way to greatly improved process control strategies. PMID- 12889835 TI - Pair-dependent co-aggregation behavior of non-flocculating sludge bacteria. AB - Two strains of non-flocculating sewage sludge bacteria (Xanthomonas sp. S53 and Microbacterium esteraromaticum S51) showed 91% and 77% co-aggregation, respectively, with Acinetobacter johnsonii S35 using a spectrophometric assay. The co-aggregates in case of Xanthomonas sp. S53 and A. johnsonii S35 were above 100 microm and stable against EDTA (2 mM) and a commercial protease (0.2 mg ml( 1)). Protease/periodate pretreatment of the partners did not affect this co aggregation. On the other hand, co-aggregates of M. esteraromaticum S51 and A. johnsonii S35 (50-70 microm) were deflocculated by EDTA or protease. Protease pretreatment of M. esteraromaticum S51 and periodate pretreatment of A. johnsonii S35 prevented their co-aggregation with respective untreated partners. The potential co-aggregation mechanisms of A. johnsonii S35 varied depending upon the other partner involved. PMID- 12889836 TI - Stereoselective reduction of 2-substituted cyclohexanones by Saccharomyces cerevisiae. AB - A comparative study of two modifications of enzymic reduction of ethyl N-[2-[4 [(2-oxo-cyclohexyl)methyl]phenoxy]ethyl]carbamate (1), an insect juvenile hormone bioanalog, was performed using Saccharomyces cerevisiae in two bioreactors of different size, 250-ml shake-flask and 1-l fermenter. The two major products of this reduction were obtained in 45-49% (w/w) yields but with > 99% enantiomeric purity. Their absolute configurations were assigned as ethyl (1S,2S)-N-[2-[4-[(2 hydroxycyclohexyl)methyl]phenoxy]ethyl]carbamate (2a) and ethyl (1R,2S)-N-[2-[4 [(2-hydroxycyclohexyl)methyl]phenoxy]ethyl]carbamate (3a). PMID- 12889837 TI - Purification of fusion ferritin from recombinant E. coli using two-step sonications. AB - Fusion ferritin, combined by heavy chain ferritin (21 kDa) and light chain ferritin (19 kDa), was expressed in recombinant E. coli. The fusion ferritin was easily purified by two-step sonications as well as gel filtration chromatography. SDS-gel electrophoresis showed a single band of 38 kDa with heavy and light chains. MALDI-TOF MS gave a molecular weight of fusion ferritin was 38 kDa. The specific activity and yield of purified fusion ferritin are 0.41 Fe3+ mg mg(-1) of protein and 66%. Those values are larger than the previous ones of 0.2 Fe3+ mg mg(-1) (Kim et al. 2001). PMID- 12889838 TI - cDNA cloning and mRNA expression of L-3,4-dihydroxyphenylalanine decarboxylase gene homologue from the silkworm, Bombyx mori. AB - L-3,4-Dihydroxyphenylalanine decarboxylase (DDC) cDNA, from Bombyx mori that contains an open reading frame of 1437 bp encoding 478 amino acids, was cloned and characterized. Expression analyses of B. mori DDC mRNA by Northern and in situ hybridization indicated that expression of silkworm DDC expression is possibly controlled by neuropeptide hormones in tissue- and stage-specific manners. PMID- 12889839 TI - Preventing osteoporotic fractures with bisphosphonates: a review of the efficacy and tolerability. AB - Although change in bone mineral density was the outcome most commonly measured in early clinical trials of osteoporosis therapies, it is now understood that the most clinically important outcome is reduction in the risk of fractures. Of currently available osteoporosis therapies, the bisphosphonates have been most thoroughly investigated in studies with fracture risk as the primary outcome. The most widely studied bisphosphonates include etidronate, alendronate and risedronate. Alendronate and risedronate have the most compelling evidence for vertebral and non-vertebral fracture reduction. This review provides a comprehensive overview of the anti-fracture efficacy of bisphosphonates at the spine, hip, and non-vertebral sites. PMID- 12889840 TI - Suicide and euthanasia in late life. AB - Epidemiological studies of suicide in the elderly indicate that, in the last few decades, there has been a relevant increase in suicide rates in old age in a number of Asian and Latin nations, with an almost parallel decrease in Anglo Saxon counties. Mental disorders, particularly depression, physical illness, personality traits such as hostility, hopelessness, the inability to verbally express psychological pain and dependency on others, recent life events and losses are all factors that may contribute to suicide in later life. Compared with suicide in other age groups, mors voluntaris in late life is associated with the use of highly lethal methods, less ambivalence and impulsivity, and more determination and intent to die. Accordingly, elderly suicidal individuals are more likely than younger subjects to complete rather than attempt suicide. Some evidence suggests also that the characteristics of elderly individuals who attempt suicide may not overlap with those who complete suicide. Death thoughts and suicidal ideations are relatively rare among mentally healthy elderly adults, and are less predominant in this age bracket. However, whether elderly suicidal behaviour exists along a continuum, progressing in severity from death thoughts and suicidal ideation to suicide attempts and completed suicide, remains unclear. Assisted suicide and euthanasia in the elderly have been associated with the desire to escape chronic physical pain and suffering caused by terminal illness, and to relieve mental anguish and feelings of hopelessness, depression and extreme "tiredness of life." The role of the family and those treating chronically ill members is crucial in the final stages of life, particularly when autonomy and the ability of the elderly individual to make end-of-life decisions are compromised. The main aspects associated with these controversial phenomena, particularly from a transcultural perspective, are reviewed in this article. PMID- 12889841 TI - Effect of six candidate genes on early aging in a French population. AB - BACKGROUND AND AIMS: The objective of this study was to examine the association between an aging indicator previously defined from a nationwide population study, and lipids and apolipoproteins, angiotensin converting enzyme, paraoxonase activities, and six candidate genes related to the aging process. METHODS: Two hundred and fifty-six healthy Caucasian men (69.8 +/- 4.0 years) were included in the study. Total cholesterol, triglycerides, HDL-cholesterol, lipoprotein(a), apolipoprotein A1, B and E concentrations, and the activities of paraoxonase, arylesterase, and angiotensin-converting enzymes were determined by standardized laboratory methods. A multiplex assay was used to genotype the studied polymorphisms: apolipoprotein E, lipoprotein lipase, paraoxonase, methylenetetrahydrofolate reductase, cystathionine beta-synthase and angiotensin converting enzymes. RESULTS: Paraoxonase polymorphism at codon 192 (Gln/Arg) was the only one significantly associated with the aging indicator, Gln homozygotes being more advanced in aging compared with Arg allele carriers. It was also observed that the aging indicator was positively correlated with serum concentrations of total cholesterol, triglycerides and apolipoprotein B, and negatively with the activities of basal and stimulated paraoxonase and arylesterase. Multiple regression analysis showed that triglycerides and basal paraoxonase activity explain 13.6% of the variance of the aging indicator. CONCLUSIONS: Triglyceride concentration and paraoxonase gene and activities may contribute to the aging process. Taking into account the smallness of the sample size, and the poor level of significance due to the im-plication of paraoxonase polymorphism at codon 192, these results need to be verified in further studies on a greater number of subjects. PMID- 12889842 TI - Pulmonary embolism in very old patients. AB - BACKGROUND AND AIMS: Diagnosis of pulmonary embolism (PE) in the elderly remains difficult and is often late, although its incidence increases steadily with age. Since few studies have reported the clinical picture of this disease in very old patients, the aim of our study was to focus on clinical, instrumental and laboratory aspects in patients 75 years of age and over with suspected PE. METHODS: Symptoms, signs, risk factors for venous thromboembolic diseases, 12 lead electrocardiograms, B-mode echocardiograms, chest X-rays, leg compression venous ultrasonography, latex-assay quantitative D-dimer, and arterial blood gas analyses were collected for 96 elderly patients of 75 years and over (mean age 81.22 +/- 4.81) with suspected PE, admitted to our acute geriatric ward from 1997 to 2000. Patients were divided into two groups: 59 patients with PE (PE+) confirmed by pulmonary angiography, scintigraphic lung scan or necropsy, and 37 patients with normal lung scan or without PE at necropsy (PE-). RESULTS: Dyspnea and chest pain were the most common symptoms in both groups, but significantly more frequent in the PE+ group. Tachycardia, fever, cyanosis, and tachypnea were the most common objective signs in both groups; tachycardia and tachypnea were not significantly different between the two groups. Bed rest, venous insufficiency and leg deep vein thrombosis were the most common venous risk factors in PE+ and significantly different with respect to PE-. Sinus tachycardia, ST-T abnormalities and right bundle branch block were the most common electrocardiographic aspects in PE+; echocardiograms showed significantly more frequent acute involvement of right heart in PE+, whereas chest X-rays were rarely compatible with PE and did not reveal differences between the two groups. D-Dimer was significantly higher in PE+ but was below 500 ng/mL only in 25% of cases in PE-. More severe hypoxemla, oxy-hemoglobin hyposaturation and increased alveolar-arterial oxygen gradients were found at arterial blood gas analysis in PE+. CONCLUSIONS: Although many analyzed clinical, instrumental and laboratory aspects were significantly more frequent in the PE+ group, they were all non specific and not conclusive for diagnosis of PE. Our retrospective study focus may add information about the picture and diagnostic difficulties of PE in very old patients. PMID- 12889843 TI - Anabolic and catabolic hormonal responses to experimental two-set low-volume resistance exercise in sedentary and active elderly people. AB - BACKGROUND AND AIMS: The influence of acute low-volume resistance exercise on serum growth hormone (GH), insulin-like growth factor I (IGF-I), dehydroepiandrosterone sulphate (DHEAS), total testosterone (TT) and cortisol was ascertained in elderly subjects. METHODS: Forty-seven independent, community dwelling volunteers aged >65 years were recruited: 23 (11 men, 12 women) were sedentary individuals, and 24 (12 men, 12 women) had been regularly involved in physical activity for several years. The protocol consisted of two sets of leg extensions: one graded by loading to reach maximal power; the other consisted of 10 consecutive leg extensions using a load corresponding to maximal power. RESULTS: IGF-I levels increased immediately after exercise, returning almost completely to pre-exercise values by the 15-minute post-exercise time point. The changes in all four study groups were similar. Not any of the groups presented systematic exercise-induced changes in circulating GH, DHEAS and TT levels. With respect to pre-exercise cortisol levels, significant decreases were observed both at the immediate and at the 15-minute post-exercise time points. These changes were independent of gender and physical activity level. CONCLUSIONS: Our data indicate that low-volume resistance exercise may decrease cortisol levels and increase serum anabolic/catabolic hormone ratios. In view of the experimental character of our resistance training protocol and the lack of control-day data, these results should be corroborated by long-term low-volume resistance training programs. PMID- 12889844 TI - Poor correlation of mid-femoral measurements by CT and hip measurements by DXA in the elderly. AB - BACKGROUND AND AIMS: Hip fracture is a devastating event in terms of outcome in the elderly, and the best predictor of hip fracture risk is hip bone density, usually measured by dual X-ray absorptiometry (DXA). However, bone density can also be ascertained from computerized tomography (CT) scans, and mid-thigh scans are frequently employed to assess the muscle and fat composition of the lower limb. Therefore, we examined if it was possible to predict hip bone density using mid-femoral bone density. METHODS: Subjects were 803 ambulatory white and black women and men, aged 70-79 years, participating in the Health, Aging and Body Composition (Health ABC) Study. Bone mineral content (BMC, g) and volumetric bone mineral density (vBMD, mg/cm3) of the mid-femur were obtained by CT, whereas BMC and areal bone mineral density (aBMD, g/cm2) of the hip (femoral neck and trochanter) were derived from DXA. RESULTS: In regression analyses stratified by race and sex, the coefficient of determination was low with mid-femoral BMC, explaining 6-27% of the variance in hip BMC, with a standard error of estimate (SEE) ranging from 16 to 22% of the mean. For mid-femur vBMD, the variance explained in hip aBMD was 2-17% with a SEE ranging from 15 to 18%. Adjusting aBMD to approximate volumetric density did not improve the relationships. In addition, the utility of fracture prediction was examined. Forty-eight subjects had one or more fractures (various sites) during a mean follow-up of 4.07 years. In logistic regression analysis, there was no association between mid-femoral vBMD and fracture (all fractures), whereas a 1 SD increase in hip BMD was associated with reduced odds for fracture of approximately approximately 60%. CONCLUSIONS: These results do not support the use of CT-derived mid-femoral vBMD or BMC to predict DXA-measured hip bone mineral status, irrespective of race or sex in older adults. Further, in contrast to femoral neck and trochanter BMD, mid-femur vBMD was not able to predict fracture (all fractures). PMID- 12889845 TI - Impact of the joint space width measurement method on the design of knee osteoarthritis studies. AB - BACKGROUND AND AIMS: Recent guidelines recommend measurement of articular loss over several years, determined by conventional X-rays, as the principal outcome measure in clinical trials of potential structure-modifying drugs in osteoarthritis (OA). The aim of this study was to assess the impact of the joint space width measurement method on sample size calculation in knee OA studies. METHODS: Standard knee X-rays were taken in 212 patients with knee OA at baseline and after 3 years of follow-up. Mean joint space width (JSW) was measured with an in-house computer-assisted method. Minimum JSW, measured with a graduated magnifying lens, was taken as external standard. After calculation of the intra- and inter-observer reproducibility of the JSW, sensitivity to change was assessed using the standardized response mean (SRM). The number of patients needed to identify a mean significant difference of 0.5 mm in joint space narrowing between the placebo and the treated group, after 3 years of follow-up, was then calculated. RESULTS: JSW measured with the computer-assisted technique showed better intra- and inter-observer reproducibility than when using the magnifying lens. JSW values measured with our computer-assisted method were significantly correlated with JSW values obtained using the magnifying lens (r=0.87, p<0.001). The SRM were 0.44 and 0.40 for the computer-assisted method and magnifying lens, respectively. The number of patients needed was 131 per group using the computer assisted method, and 104 using the magnifying lens. CONCLUSIONS: Our method of measurement of JSW may be of potential use in longitudinal studies evaluating the effect of structure-modifying drugs in OA, due to its high level of precision and efficiency. However, although sensitivity to change is markedly better with the digitized method compared with the graduated magnifying lens, we recommend the measurement of mean and minimum JSW in structure-modifying OA trials. PMID- 12889846 TI - Prediction of falls among older people in residential care facilities by the Downton index. AB - BACKGROUND AND AIMS: Falls are frequent among older people living in residential care facilities. The aim of this study was to investigate the prediction accuracy of the Downton fall risk index among older people living in residential care facilities at 3, 6 and 12 months, and with two different definitions of falls. METHODS: Seventy-eight residents in one residential care facility, 56 women and 22 men, mean +/- SD age 81 +/- 6 years, participated in this study. Forty-seven percent of participants had dementia, 45% depression, and 32% previous stroke. Forty-one percent of participants used a walking device indoors, and the median score of the Barthel ADL Index was 16. At baseline, the Downton fall risk index was scored for each individual. A score of 3 or more was taken to indicate high risk of falls. Participants were followed up prospectively for 12 months, with regard to falls indoors. RESULTS: At 3, 6 and 12 months, and using a fall definition including all indoor falls, sensitivity ranged from 81 to 95% with the highest value at 3 months, and specificity ranged from 35 to 40%. The prognostic separation values ranged from 0.26 to 0.37. Within 3 months, the risk of falling was 36% in the high-risk group (index score > or = 3) and 5% in the low-risk group. The accuracy of predictions did not improve when applying a fall definition in which falls precipitated by acute illness, acute disease, or drug side-effects were excluded. CONCLUSIONS: Already after 3 months, the Downton fall risk index appears to be a useful tool for predicting falls, irrespective of their cause, among older people in residential care facilities. PMID- 12889847 TI - Nutritional assessment of demented patients: a descriptive study. AB - BACKGROUND AND AIMS: It is well-known that subclinical and overt malnutrition are strong predictive indices of morbidity and mortality in old subjects, particularly in demented ones, and may deeply affect the quality of life. The aim of this study was evaluation of nutritional status in demented patients, as a whole and according to type of dementia, at the moment of hospital admission and before discharge. METHODS: The study concerns 174 old demented patients, aged 80.2 +/- 8 SD, diagnosed as having Alzheimer's disease, vascular dementia, degenerative and vascular dementia, reversible dementia, and other types of dementia. In each subject, anthropometric measures, body composition analysis by the bioelectric impedance technique, Mini Nutritional Assessment (MNA) and cognitive, functional and affective assessment by conventional geriatric instruments were performed. A blood sample allowed assay of the main biochemical nutritional markers. At the end of the hospitalization period, the same cognitive, functional and nutritional assessment was repeated, in order to evaluate the effects of vitamin and protein supplements and of care during meals. RESULTS: In all subgroups of demented patients, obtained according to type of dementia, the mean MNA score was indicative of risk for malnutrition. Furthermore, the MNA score was significantly related to severe cognitive impairment, functional status, comorbidity, BMI values, and transferrin and total protein serum levels. Malnourished patients and demented elderly at risk for malnutrition (according to the MNA score) were given oral nutritional supplements during hospitalization, lasting a mean of 45 days. Before discharge, these two subtypes of demented patients showed substantial maintenance of their cognitive, functional and nutritional status, whereas the subgroup of well-nourished demented patients exhibited significant worsening of the nutritional pattern. CONCLUSIONS: Demented patients show a high percentage of malnutrition, particularly evident in subjects with deeper cognitive impairment. Nutritional status seems to be linked more to functional abilities than to duration of disease. However, nutritional intervention or special care during meals may act in synergy with specific pharmacologic therapy of dementia. PMID- 12889848 TI - Medical history, cognitive status and mobility at the age of 90. A population based study in Tampere, Finland. AB - BACKGROUND AND AIMS: The oldest-old population is expanding rapidly. There is a new need for clinical information about this group, which is actively using social and health care. We studied the population of people born in 1907-1910 and living in the city of Tampere (Finland) at the age 90 (N=916, 79.4% women, 20.6% men); 71.7% of the population lived in the community and 28.3% in institutions. METHODS: Medical records of 832 (90.8%) nonagenarians were obtained. We registered diagnoses of chronic diseases or diseases that required hospitalization at any time of their lives, as well as physicians' notes on their memory and mobility. Diseases were coded and grouped according to ICD-10. RESULTS: The most common diagnosis groups were cardiovascular diseases (78.3%), gastrointestinal diseases (58.6%), infections (53.6%) and trauma (49.6%). There was an average of 8 chronic or severe diseases mentioned in patient records. The diagnosis of dementia was mentioned in 26.7% of cases, most of them living in institutions; a problem with memory--from forgetfulness to dementia--was mentioned in 35.9% of cases; 37.5% were able to move using no or a light support, 8.3% were bedridden. CONCLUSIONS: We conclude that this age group suffers from numerous chronic diseases influencing mobility and cognition. Dementia seems to be the most important symptom leading to institutionalization. PMID- 12889849 TI - The influence of differing social ties on decline in physical functioning among older people with and without chronic diseases: the Longitudinal Aging Study Amsterdam. AB - BACKGROUND AND AIMS: Global social support measures have been shown to be related to several health outcomes. However, little is known about the effects of differing social ties and their support on the risk for decline in physical functioning among older people, without as compared with those with chronic diseases. This study examines whether differing types of social ties and support differentially mitigate the negative effects of chronic diseases on decline in physical functioning. METHODS: Using data from two cycles of the Longitudinal Aging Study Amsterdam (N=2357), logistic regression analyses adjusted for baseline functioning, age, gender, and incidence of chronic diseases were conducted, to assess the effect of differing social ties for subgroups with different numbers of chronic diseases. Information about the presence of differing social ties included partner status and numbers of daughters, sons, other family members, and non-kin relationships. Social support included instrumental and emotional support, and the experience of loneliness. Decline in physical functioning was determined by substantial change after three years on a 6-item self-report scale. RESULTS: Although having a partner had a protective effect on decline in physical functioning in people without chronic diseases at baseline, this was not the case for those with chronic diseases. Total network size had an adverse effect in older people without chronic diseases, but a positive effect when chronic diseases were present, mainly due to a positive effect of the number of daughters and non-kin relationships. CONCLUSIONS: Our results provide evidence that differing types of social relationships and the support they provide, differentially influence decline in physical functioning in older people, with or without chronic diseases. PMID- 12889850 TI - Income differentials in functional disability in old age: relative risks of onset, recovery, decline, attrition and mortality. AB - BACKGROUND AND AIMS: Socioeconomic status (SES) differences in health decline in late life may be underestimated, because the relatively higher risks of attrition of lower-SES persons are seldom taken into account. This longitudinal study aimed at comparing income differences in the course of disability, non-mortality attrition and mortality in older adults. METHODS: A sample population of 3107 older adults who participated in the 1992/1993 baseline of the Longitudinal Aging Study Amsterdam was examined regarding changes in functional disability in 1998/1999. SES was indicated by household income. RESULTS: Multinomial regression analyses revealed that, for men without disability at baseline, the relative rate for attrition was four times higher and the mortality rate was twice as high for low-income vs high-income persons. For non-disabled women, the relative risk for the onset of disability was nearly twice as high for low-income vs high-income persons. For both men and women, these risks decreased only slightly when behavioral and psychosocial risk factors were taken into account. Among persons with disability at baseline, the relative risks for attrition (for women) and mortality (for men) were twice as high for low-income persons, but no income differences were found with respect to recovery and decline. Adjustment for risk factors decreased the relative risks for attrition and mortality to a non significant level. CONCLUSIONS: Income inequality in health in late life is to a large degree explained by the higher incidence of disability among lower-status women and by the higher attrition and mortality risks among lower-status men. PMID- 12889851 TI - Gastric myoelectrical activity in patients with recurrent gastric or duodenal ulcers. AB - The aim of this study was to characterize gastric myoelectrical activity in patients with recurrent gastric ulcer (GU) or duodenal ulcer (DU), and to compare gastric motility between these two groups of patients. Studies were performed in 59 patients with recurrent active peptic-ulcer disease as diagnosed by gastrointestinal endoscopy: 31 patients had a GU and 28 patients had a DU. Gastric myoelectrical activity was evaluated by cutaneous electro-gastrography (EGG). The following EGG parameters were assessed: the percentage of normogastria (regular 2.4-3.6 cpm slow waves); the EGG power ratio; and the occurrence of a postprandial dip (PD), which is the transient decrease in EGG frequency after a meal. In the GU group, no significant change occurred in the percentage of normogastria or in the EGG power ratio observed after treatment with a proton pump inhibitor. During the healed stage, the occurrence of PD remained unchanged. In contrast, in the DU group, the percentage of normogastria and the EGG power ratio were significantly increased after treatment. Moreover, during the healed stage, the occurrence of PD significantly increased compared with that during the active stage. These findings suggest that abnormal gastric myoelectrical activity plays an important role in the pathophysiology of recurrent GU rather than DU. PMID- 12889852 TI - Neural control of the internal anal sphincter motility. AB - Control mechanism of smooth muscle movement of the internal anal sphincter (IAS) by enteric and extrinsic neuvous systems in the dog was investigated. Responses of IAS muscle strips to electrical field stimulation (EFS) and neurotransmitter agents were recorded in vitro. The contraction response to norepinephrine or to EFS was inhibited by phentolamine. The relaxation induced by EFS was not affected by phentolamine, propranolol or atropine. The mechanical activity of smooth muscle in colon and anorectum during spontaneous defecation was recorded using strain gauge force transducers. The colon and anorectum showed the characteristic motility pattern during defecation: 1) The giant migrating contraction of the colon propagated to the rectum, 2) The relaxation of the rectum prior to the contraction, and 3) The IAS muscles continued to relax while the giant contractions of the colon were migrating to the rectum. Sacral nerves were stimulated electrically and the responses of smooth muscles in the rectum and IAS were recorded. The sacral nerve stimulation induced a relaxation followed by contraction of smooth muscle in the rectum and the relaxation in IAS. The mechanical responses of smooth muscle in the IAS were modulated by alpha adrenergic excitatory and non-adrenergic, non-cholinergic inhibitory nerves. During defecation, the relaxation of IAS smooth muscle was associated with a characteristic motility pattern of the colon and anorectum. The enteric nervous systems may be organizing the motility of these muscles by way of the motor neurones under the control the extrinsic nervous systems. PMID- 12889853 TI - Glycolytic ATP production regulates muscarinic cation currents in guinea-pig ileum. AB - We investigated the possible sources of intracellular ATP which was previously shown essential for maintaining the muscarinic cationic channel activities (or currents; I(cat)) in guinea-pig ileal myocytes, using two variants of patch clamp techniques. Deprivation of external glucose or its replacement with 2 deoxyglucose significantly reduced the magnitude of I(cat), recorded with nystatin-perforated method, with greater efficacy than for voltage-dependent Ca2+ current Intracellular dialysis of ileal myocytes with key substrates for glycolysis, oxidative metabolism and creatine-phosphocreatine system all resulted in a comparably effective maintenance of I(cat), which was abolished by inhibitors for these ATP-producing systems, 3-bromopyruvate, cyanide and 2,4 dinitrofluorobenzene (DNFB), respectively. However, amongst these inhibitors, only 3-bromopyruvate effectively reduced I(cat) recorded with the nystatin perforated method. These results strongly suggest the exclusive physiological importance of glycolytic ATP production in maintaining I(cat), activity, and thus this mechanism may play a role in the regulation of gut motility. PMID- 12889854 TI - Stem cells and regenerative medicine. AB - Stem cells have been defined as clonogenic cells that undergo both self-renewal and differentiation to more committed progenitors and functionally specialized mature cells. Of late years, stem cells have been identified in a variety of tissues of an adult body. Depending on the source, they have the potential to form one or more, or even all cell types of an organism. Stem cell research provided some outstanding contributions to our understanding of developmental biology and offered much hope for cell replacement therapies overcoming a variety of diseases. The establishment of human ES cell lines enabled us to generate all tissues we comprise. Recently, excitement has been evoked by the controversial evidence that adult stem cells have a much higher degree of developmental plasticity than previously imagined. More recently, the existence of multipotent somatic stem cells in bone marrow has been reported. Combined with these discoveries and achievements as well as the developing technologies, scientists are now trying to bring stem cell therapies to the clinic. PMID- 12889855 TI - Establishment and characterization of a new human cell line (EJ) derived from endometrial carcinoma. AB - We present a new cell line, EJ established from an invasive endometrioid adenocarcinoma of the uterine corpus in a 56-year-old patient. The cells show rapid growth in culture with a doubling time of 16 h and high migration activity. Monolayer-cultured cells were polygonal in shape showing a tendency to pile up without contact inhibition. Subcutaneous transplantation of the EJ cells into nude mice formed solid tumors that were histologically diagnosed as adenocarcinoma, whereas no metastasis was observed. Cultured EJ cells produced tissue polypeptide antigen (IPA). Genetic and molecular analyses revealed high telomerase activity but not estrogen receptor alpha expression. Using the DNA sequencing technique, we have screened EJ cells for p53 mutation in exon 5 to 8 but no mutation of p53 was observed. This cell line appears to represent the development of a more malignant clone with divergent receptor function and growth behavior, and provides us with an interesting new tool for the study of tumorigenesis in the human endometrium. PMID- 12889856 TI - Establishment and characterization of two squamous cell carcinoma cell lines (HYVC and HMVC) derived from vulva. AB - Two cell lines, HYVC and HMVC, were established from cultures of the squamous cell carcinoma removed from the vulva of females of 37 and 46 years old, respectively. These cell lines were very similar in their biological characteristics, such as morphology (polygonal), growth pattern (32-43 hr of population doubling time, 50-25% of plating efficiency), heterotransplantability (1 x 10(7) cells produced squamous cell carcinomas in the nude mice), genetics (75-78 of modal chromosomal number), and producing the tumor markers (SCC and TPA). The HPV-DNA was not detected in either the HYVC or HMVC lines by using L1 PCR methods, however, it was detected in the HYVC using E6/E7-PCR. PMID- 12889857 TI - Establishment and characterization of new B-cell precursor leukemia cell line NALM-35. AB - A novel B-cell precursor (BCP) acute lymphoblastic leukemia (ALL) cell line, NALM 35, was established from the peripheral blood of a 40-year-old woman at diagnosis of ALL. Imunophenotyping showed BCP type III characteristics including expression of TdT, CD10, CD19, CD22, CD79a and HLA class II. T-cell and myeloid-associated antigens tested were negative except CD5 and CD28. The surrogate light chains CD179a and CD179b were positive. NALM-35 cells have the morphological appearance of lymphoblasts. Cytogenetic analysis of NALM-35 revealed an abnormal karyotype with 46, XX, add(9)(p11). Southern blot analysis of the immunoglobulin genes status of NALM-35 at 10 months after establishment showed germ line configuration of the kappa and lambda light chain genes, and rearrangement of the mu heavy chain gene. DNA fingerprinting, chromosomal analysis and immunophenotyping proved that NALM-35 was clonally derived from the primary leukemia cells. The established cell line may provide a useful model system and unprecedented opportunities for analyzing the multitude of biological aspects of normal and neoplastic B-lymphocytes and their precursors. PMID- 12889858 TI - Development of reconstituted embryos derived from transgenic embryonic stem cell nuclei. AB - This study was carried out to transform embryonic stem (ES) cells and to produce the reconstituted embryos derived from transgenic ES cell nuclei. Then, in vitro/vivo developmental potency of transgenic ES cells were compared to that of control ES cells (non-transgenic ES cells) in the reconstituted embryos. Unfertilized B6D2F1 ooplasm at metaphase II (M II) and two kinds of ES cell lines, 129SV and transgenic (tg) 129SV transformed by EGFP gene, were used as nuclear recipients and nuclear donors, respectively. The M II chromosome-spindle complex was aspirated into the pipette with a minimal volume of ooplasm. After enucleation, the ES cell nuclei was injected into the enucleated ooplasm directly. Then, reconstituted embryos were activated in SrCl2, and they were cultured in HTF medium. There was no difference of developmental rate between reconstituted embryos derived from the control (non-transgenic) and the tg ES cells. From this result, we indicated that transgenic ES cells might not change the property of peculiarity of the ES cell by gene transfer. The expression of GFP gene in the embryos was observed by fluorescence microscope at the 4-cell and more stage. As comparison with development of the embryos derived from the control and tg ES cells, the difference of the development could not be confirmed between the two cell groups. When the reconstituted embryos derived from the control and tg ES cells were transferred into oviduct or uterus of pseudopregnant females, fetuses were observed 13.5 days post coitum. However, in all fetuses, developmental arrest and regression were seen 19.5 days post coitum. PMID- 12889859 TI - Establishment of a novel method for cryopreservation and thawing of the mouse ovary. AB - During cryopreservation of ovarian tissue, the conditions of freezing and thawing are big factors controlling the survival rate of oocytes obtained. However, the conditions and procedures as they pertain to ovarian follicles and oocytes have not been established. Thus, we tried to determine the appropriate freeze-thaw times using the vitrification method with ethylene glycol and DMSO as cryoprotective agents and dd Y female mouse ovaries. The maturity rate from GV to the metaphase-II (MII) stage was 62.8% with ethylene glycol and 69.3% using DMSO, while the controls (GV oocytes obtained from a fresh ovary) showed a maturation rate of 83.6% (46/55). MII oocytes obtained by culturing GV oocytes in vitro showed a 64.3% (18/28) fertility rate via in vitro fertilization and a developmental rate into a 2 cell stage embryo of 35.7% (10/28) and into a 4-cell stage, 7.1% (2/28). However, development beyond the 8 cell stage embryo was not observed. A significant difference was not recognized between control (fresh) and ovarian tissues that had been frozen/thawed with respect to their ability to produce hormones. It is concluded that the vitrification method was effective for both freezing ovarian tissues and preserving its functional ability (maturation and capacitation). PMID- 12889860 TI - Thoracoscopic anterior instrumentation and fusion for idiopathic scoliosis. AB - Thoracoscopically assisted surgery is a new approach to access the anterior spine to perform biopsies, anterior releases, diskectomies, and anterior instrumentation and fusion for idiopathic thoracic scoliosis. This approach compromises the chest wall less than an open thoracotomy does because it uses several small portal incisions. It has been suggested that this approach allows fusion of fewer motion segments and better correction of curvature than does posterior spinal fusion and instrumentation. The technique, which is still evolving, is technically demanding, requiring advanced training and special instrumentation and anesthesia techniques. PMID- 12889861 TI - Spinal manipulative therapy for low back pain. AB - Growing interest in complementary and alternative medicine in the United States has been paralleled by increased use of spinal manipulative therapy in an attempt to manage symptoms of low back pain, spinal stenosis, and spondylolisthesis. Chiropractors have been the main practitioners of spinal manipulative therapy, with osteopaths and physical therapists providing a smaller fraction of these services. Theories explaining the mode of action of spinal manipulative therapy are largely preliminary and have focused on the mechanical effects of manipulative forces on the spine and neurologic responses to manipulation. The effects of spinal manipulation on patients with both acute and chronic low back pain have been investigated in randomized clinical trials. Most reviews of these trials indicate that spinal manipulative therapy provides some short-term benefit to patients, especially with acute low back pain. PMID- 12889862 TI - Extensor mechanism failure associated with total knee arthroplasty: prevention and management. AB - Extensor mechanism complications are the most commonly reported reasons for revision surgery after total knee arthroplasty and are a frequent source of postoperative morbidity. Patellofemoral instability is the most commonly reported extensor mechanism complication and has multiple etiologies, including prosthetic malalignment and soft-tissue imbabalce. Patellar fracture or rupture of either the quadriceps or patellar tendon can cause catastrophic disruption of the extensor mechanism. Although some stable fractures can be successfully managed nonsurgically, displaced fractures or tendon rupture often lead to poor results. Other complications include patellar clunk and soft-tissue adhesions, prosthetic wear or loosening, and osteonecrosis. Increased understanding of implant alignment, rotation, and soft-tissue balance, as well as improved design of the trochlear groove of femoral implants and patellar components, has resulted in a decline in extensor mechanism complications. Appropriate prosthetic selection and meticulous surgical technique remain the keys to avoiding unsatisfactory results and revision surgery. PMID- 12889863 TI - Magnetic resonance imaging of the pediatric spine. AB - Magnetic resonance is an excellent modality for imaging the pediatric spine. Its successful use requires understanding both the basic physics and the sedation protocols necessary for acquiring high-resolution images. Interpreting the images accurately depends on appreciating the differences between the normal anatomy of the pediatric and the adult spine. Evaluating the images requires familiarity with the differential diagnosis of pediatric spine disease, including the most common processes (infections, neoplasms, and trauma) as well as spinal dysraphism. Despite the acknowledged usefulness of magnetic resonance imaging of the pediatric spine, controversies remain related to its safety in this age group and its limitations in diagnosing and evaluating scoliosis and tethered cord syndrome. PMID- 12889864 TI - Injury to the tarsometatarsal joint complex. AB - Tarsometatarsal joint complex fracture-dislocations may result from direct or indirect trauma. Direct injuries are usually the result of a crush and may involve associated compartment syndrome, significant soft-tissue injury, and open fracture-dislocation. Indirect injuries are often the result of an axial load to the plantarflexed foot. Midfoot pain after even a minor forefoot injury should raise suspicion; up to 20% of tarsometatarsal joint complex injuries are missed on initial examination. An anteroposterior radiograph with abduction stress may reveal subtle injury, but computed tomography is the preferred imaging modality. The goal of treatment is the restoration of a pain-free, functional foot. The preferred treatment is open reduction and internal fixation, using screw fixation for the medial three rays and Kirschner wires for the fourth and fifth tarsometatarsal joints. Satisfactory outcome can be expected in approximately 90% of patients. PMID- 12889865 TI - Chronic exertional compartment syndrome. AB - Chronic exertional compartment syndrome is an often overlooked and uncommon cause of pain in the extremities of individuals who engage in repetitive physical activity. A thorough history, a careful physical examination, and compartment pressure testing are essential to establish the diagnosis. Catheter measurements can provide useful information on baseline resting compartment pressures as well as compartment pressures after exercise or trauma. Patients with chronic exertional compartment syndrome usually do not respond to nonsurgical therapy other than completely ceasing the activities that cause the symptoms. Surgical intervention entails fasciotomies of the involved compartments. Although obtaining accurate compartment pressure measurements can be difficult and fascial releases must be done carefully, patients typically have satisfactory functional results and are able to return to their usual physical activities after fasciotomy. PMID- 12889866 TI - Proximal row carpectomy and intercarpal arthrodesis for the management of wrist arthritis. AB - For advanced noninflammatory wrist arthritis, the most common surgical treatments to preserve motion are proximal row carpectomy and scaphoid excision with capitohamate-lunotriquetral arthrodesis. Both procedures have documented successful outcomes. Proximal row carpectomy is simpler but typically is contraindicated when degeneration of the capitate head cartilage exists. Scaphoid excision with capitohamate-lunotriquetral arthrodesis is more complex but may provide greater grip strength and can be successful in the presence of capitate degeneration. Treatment selection should be based on surgeon preference and experience as well as on the patient's understanding of the possible complications and benefits of each procedure. PMID- 12889867 TI - Metastatic bone disease of the humerus. AB - Metastatic bone disease is the most common cause of destructive bone lesions in adults, and involvement of the humerus is common. Patients with destructive lesions involving <50% of the cortex are treated nonsurgically with external beam irradiation. Patients with diaphyseal lesions involving > or =50% of the cortex or those with pain after irradiation can be treated with intramedullary nailing to achieve rigid fixation. Although closed intramedullary nailing is used most often, open nailing with methylmethacrylate is appropriate for destructive lesions in which rigid fixation cannot be achieved with closed nailing. Plate fixation is acceptable when adequate proximal and distal cortical bone is present for screw purchase, although proximal humeral lesions usually are treated with prosthetic arthroplasty. Postoperative external beam irradiation can help prevent disease progression and subsequent loss of fixation. However, when disease progression persists or rigid internal fixation is not feasible because of extensive bone destruction, wide resection and reconstruction with a custom prosthesis can be done. PMID- 12889868 TI - NY-ESO-1 mRNA expression and immunogenicity in advanced prostate cancer. AB - NY-ESO-1 mRNA expression was investigated in advanced prostate cancer by conventional and quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR). NY-ESO-1 mRNA was detected in 20 of 53 (38%) tumor specimens. Four of 15 (27%) stage C, 1 of 3 stage D1 (33%) and 15 of 35 (43%) stage D2 prostate cancers were positive. The presence of NY-ESO-1 antibodies was evaluated in sera from a panel of 218 patients with prostate cancer, including the 53 patients whose tumors were examined for NY-ESO-1 mRNA expression. NY-ESO-1 antibodies were detected in 1 of 30 (3.3%) stage D1 and 9 of 110 (8.2%) stage D2 patients, whereas none of the 78 patients with localized prostate cancer (stages A, B and C) had detectable NY-ESO-1 antibodies. Of the 53 patients whose tumors were examined for NY-ESO-1 mRNA expression, 2 of 20 patients with NY-ESO-1 mRNA positive tumors had NY-ESO-1 antibodies. No antibody was found in the sera of 32 patients with NY-ESO-1 mRNA-negative tumors, with the exception of one patient with regional lymph node metastasis (stage D1). CD8 T cell responses specific to NY-ESO-1 were detected in two of three patients with NY-ESO-1 antibodies. PMID- 12889869 TI - Alternating conjugated and transannular chromophores: tunable property of fluorene-paracyclophane copolymers via transannular pi-pi interaction. AB - [structure-see text]A series of fluorene-dithia[3.3]paracyclophane copolymers was synthesized by Suzuki coupling reactions of suitable precursors. Transannular pi pi interactions altered the electronic and optical properties of the polymer backbone significantly. A large red shift in the emission spectrum of the unsubstituted polymer was accompanied by enhanced photoluminescence (PL) efficiency. Substitution in the cyclophane unit resulted in PL quenching. PMID- 12889870 TI - Emission color tuning in AlQ3 complexes with extended conjugated chromophores. AB - [reaction: see text] A new method for the synthesis of 5-arylethynyl-8 hydroxyquinoline ligands using Sonogashira-Hagihara coupling was developed. The electronic nature of arylethynyl substituents affects the emission color and quantum yield of the resulting Al(III) complex. Photophysical properties of the metallocomplexes correspond to the electron-withdrawing/-donating character of the arylethynyl substituents. Optical properties of such Al(III) complexes correlate with the Hammett constant values of the respective substituents. This strategy offers a powerful tool for the preparation of electroluminophores with predictable photophysical properties. PMID- 12889871 TI - Fused triazoles via tandem reactions of activated Cinchona alkaloids with azide ion. Second Cinchona rearrangement exemplified. AB - [reaction: see text] Intramolecular 1,3-dipolar cycloadditions of cinchona azides to the C10-C11 alkyne and C10-C11 olefin unit of the alkaloid have been designed via tandem strategy. A variety of fused triazoles and triazolines with a bis azahomotwistane skeleton have been prepared. In trifluoroethanol, O mesylcinchonidine 7-OMs and NaN(3) furnish triazole 8 as well as cage-expanded 1,5-diazatricyclo[4.4.1.0(3,8)]undecane derivative 10. Both fused triazoles 8 and 10 are formed with retention of configuration at C9 and C3, respectively. 1 Azabicyclo[3.2.2]cage expansion is shown to be reversible. PMID- 12889872 TI - Diverging stereochemical pathways in an intramolecular Diels-Alder reaction determined by dienophile structure. AB - [reaction: see text] Intramolecular Diels-Alder cycloaddition of tolylsulfonyl alkynone 5a and vinyl ketone 19 led to single isomers with opposite diastereoselectivity. Decalin 20 incorporates the correct C(9)-C(11) stereochemical array for a projected synthesis of HMP-Y1 (1). PMID- 12889873 TI - Variable ion selectivity in [N.3.3](1,3,5)crownophanes: the "breathing" process. AB - [reaction: see text] A series of dithia[n.3.3](1.3.5)crownophanes were synthesized by cesium carbonate-assisted intramolecular cyclization reactions in 10-31% yields. Their complexation behavior exhibits an unusual ion-selectivity preference due to the presence of a "breathing" process of the dithia[3.3]metacyclophane moiety, which indirectly controls the ion selectivity through the adjustment of the cavity size of the crown unit. This breathing mechanism is also supported by observation made in X-ray crystallographic analysis. PMID- 12889874 TI - Synthesis of muscothiazoles A and B: Critical role of methyl group substitution in RCM-based syntheses of macrocycles. AB - [reaction: see text] Muscothiazoles A (2b) and B (2c) have been prepared by two approaches that differ in the order of assembly of the rings. Comparative studies show that substitution of the carbon chains in substrate 5 or 12 (respective precursors to 13-membered and 14-membered rings by RCM), even by a single methyl group, can have a profound effect on increasing the efficiency of the macrocyclization. PMID- 12889875 TI - An efficient synthesis of vinylsilanes from acylsilanes and alkyl 1-phenyl-1H tetrazol-5-yl sulfones. Brook vs Smiles rearrangement. AB - [reaction: see text] Vinylsilanes are formed in high yields in the reaction of representative acyl(trimethyl)silanes with anions generated from Kocienski's sulfones. PMID- 12889876 TI - A new class of spermidine-derived alkaloids. AB - [structure: see text] Four new alkaloids, dovyalicins A-D, were isolated from Dovyalis macrocalyx. Their structures were established by two-dimensional COSY, NOESY, HSQC, and HMBC NMR experiments as well as chiroptical methods. The compounds, possessing spermidine as part of a perhydro-1,5-diazocine or a perhydro-1,4-diazepine moiety, constitute a new group of polyamine-derived alkaloids. PMID- 12889877 TI - Total syntheses of bioactive oxidized ethanolamine phospholipids. AB - [reaction: see text] Truncated ethanolamine phospholipids containing aldehyde functionality, e.g. OVPE, and the corresponding acids, are generated by oxidative cleavage of polyunsaturated phospholipids. To confirm their identities and facilitate studies of the chemistry and biological actions of these analogues of biologically active phosphatidylcholines, e.g. OVPC, total syntheses were developed. An efficient general strategy was used that features selective N protection of 2-lysophosphatidylethanolamine, and generation of the target compounds by mild deprotection of stable precursors. PMID- 12889878 TI - Synthesis of a nitrogen analogue of sphingomyelin as a sphingomyelinase inhibitor. AB - [structure: see text] Sphingomyelin nitrogen analogue 1 was designed and synthesized as a sphingomyelinase inhibitor. The synthesis was established by continuous Hofmann rearrangement and Crutius rearrangement as key steps in constructing the 3-hydroxy-1,2-diamine structure in the backbone of 1. This analogue showed moderate inhibitory activity toward SMase isolated from B. cereus. PMID- 12889879 TI - Atroposelective biaryl coupling with chiral catalysts: total synthesis of the antileishmanial naphthylisoquinoline alkaloids ancistrotanzanine B and ancistroealaine A. AB - [reaction: see text] The first total synthesis of the naphthylisoquinoline alkaloid ancistrotanzanine B and its atropo-diastereomer, ancistroealaine A, is described. The key step is the construction of the rotationally hindered and thus stereogenic biaryl axis by Suzuki coupling. While only weak internal asymmetric inductions by the stereogenic center in the dihydroisoquinoline part were observed, much better atropisomeric ratios in favor of ancistrotanzanine B were achieved by the use of chiral catalysts. Both alkaloids, in particular ancistrotanzanine B, show high antileishmanial activities. PMID- 12889880 TI - Studies on the intramolecular C[bond]H...X (X = O, S) interactions in (S)-N-acyl- 4-isopropyl-1,3-thiazolidine-2-thiones and related 1,3-oxazolidin-2-ones. AB - [reaction: see text] Two intramolecular C[bond]H...X (X = O, S) interactions in (S)-N-acyl-4-isopropyl-1,3-thiazolidine-2-thiones are documented through crystallographic and spectroscopic evidence and high-level theoretical calculations. The key role played by the sulfur atoms has been made clear by comparison with structurally related (S)-N-acyl-4-isopropyl-1,3-oxazolidin-2 ones. PMID- 12889881 TI - Silylative carbocyclization of allenyl-carbonyl units with Et3SiH catalyzed by rhodium: cis-stereoselective synthesis of homoallylic cycloalkanols. AB - [reaction: see text] A new procedure for the synthesis of cis-2 triethylsilylvinyl-cyclopentanols and -cyclohexanols from allenyl-aldehydes and ketones with Et(3)SiH through rhodium-catalyzed silylative carbocyclization is described. The use of Rh(acac)(CO)(2) (1 mol %) to promote the reaction results in a mild and convenient protocol for the three-component assembly. PMID- 12889882 TI - Synthesis of dityrosine cross-linked peptide dimers using the Miyaura-Suzuki reaction. AB - [reaction: see text] Since peroxidase-catalyzed dityrosine formation is inefficient for peptides, we have developed alternative conditions for intermolecular dityrosine formation using the Miyaura-Suzuki reaction. A one-pot reaction is effective for cross-linking short peptides, but longer peptides inhibit the Suzuki step, mandating a traditional two-step procedure using potassium acetate for the Miyaura reaction and potassium carbonate for the Suzuki coupling. These palladium-based methods are complementary to the well-established peroxidase-catalyzed oxidative phenolic coupling of full-length proteins. PMID- 12889883 TI - The total synthesis and stereochemical revision of yanucamide A. AB - [reaction: see text] The first total synthesis of yanucamide A is reported via amide and ester couplings of the key components. This synthesis has established the configuration at the previously ambiguous 3-position, and also revised the stereochemistry at the 22-position, to give 3S,12S,17S,22S for the natural product. PMID- 12889884 TI - Total Synthesis of (+)-okaramine J featuring an exceptionally facile N-reverse prenyl to C-prenyl aza-Claisen rearrangement. AB - [reaction: see text] The convergent total synthesis of (+)-okaramine J was achieved in a longest linear sequence of 12 steps from l-tryptophan tert-butyl ester. A key reaction was the acid-catalyzed room-temperature aza-Claisen rearrangement of a N-reverse-prenylated hexahydro[2,3-b]pyrroloindole to a C prenylated derivative. PMID- 12889885 TI - Bartoli indole synthesis on solid supports. AB - [reaction: see text] Bartoli indole synthesis has been performed for the first time on solid supports. Starting from Merrifield resin, immobilization of five nitro benzoic acids was performed. Addition of four different alkenyl Grignard reagents and basic cleavage leads to substituted methyl indole carboxylates in excellent purities. Features of this reaction are the stability of halide groups, ester moieties, and tolerance of o,o'-unsubstituted nitro resins. Heck and Sonogashira reactions are also possible with immobilized indoles. PMID- 12889886 TI - Diels-Alder reactions of fused pyran-2-ones with maleimides: efficient syntheses of benz[e]isoindoles and related systems. AB - [reaction: see text] The Diels-Alder reaction of some substituted 5,6,7,8 tetrahydro-2H-1-benzopyran-2-ones (1a-f) with N-substituted maleimides (2a-c) leading to fused isoindole derivatives (5a-n, 7) or, in a few cases, to bridged double cycloadducts (fused bicyclo[2.2.2]octene derivatives) (6a-f) is presented. When X = CO, the first efficient, substituent-driven aromatization of an intermediary-formed cycloadduct was observed, resulting in substituted benz[e]isoindoles (5a-k). The same type of aromatization can also be achieved in an unprecedented catalysis with Rh/C. PMID- 12889887 TI - Synthetic studies directed toward the assembly of the C-glycoside fragment of the telomerase inhibitor D8646-2-6. AB - [reaction: see text] Construction and characterization of the C-glycosidic moiety of telomerase inhibitor D8646-2-6 (1) are described. This is the first example of the C-glycosylation using electron-poor aromatics, 4-hydroxypyrone, as a glycosyl acceptor. The glycosylation reaction and base-promoted isomerization affords desired beta-C-glycoside in a 61% overall yield. PMID- 12889888 TI - "Formal" ruthenium-catalyzed [4+2+2] cycloaddition of 1,6-diynes to 1,3-dienes: formation of cyclooctatrienes vs vinylcyclohexadienes. AB - [reaction: see text] A new "formal" Ru-catalyzed [4+2+2] cycloaddition of 1,6 diynes to 1,3-dienes giving conjugated 1,3,5-cyclooctatrienes and vinylcyclohexadienes is described. This formal cycloaddition is really a tandem process, the Ru(II)-catalyzed formation of (Z)-tetraenes or vinyl-(Z)-trienes followed by a pure thermal conrotatory 8 pi- or disrotatory 6 pi electrocyclization. The proposed mechanism allows the differences in product ratio to be explained in terms of steric and stereochemical considerations. PMID- 12889889 TI - Synthesis, structure, and chemical property of the first fluorine-containing porphycene. AB - [structure: see text] A pyrrolic macrocycle, beta tetrakis(trifluoromethyl)porphycene, is the first example of a fluorine containing porphycene. Four electron-withdrawing CF(3) substituents provide a highly distorted structure and an attractive electron-deficient nature for the porphycene framework. From the electrochemical study, it is found that the LUMO energy level for the beta-trifluoromethylporphycene is 1.24 V more stabilized compared to that for etioporphyrin. Moreover, the deprotonation of the inner N[bond]H proton in the porphycene was observed upon the addition of DBU. PMID- 12889890 TI - An efficient synthesis of the C-23 deoxy, 17 alpha-hydroxy South 1 hemisphere and its cephalostatin 1 analog. AB - [reaction: see text] Methods for functionalization of the D ring of diene 1 were investigated. This study led to efficient syntheses of 3 and the subsequent 23' deoxy,17'alpha-hydroxy cephalostatin 1 analog (4). The bioactivity data of 2 and 4 are in the low nanomolar range for a 10-cell-line minipanel. PMID- 12889891 TI - A new protocol for in situ dioxirane reactions: stoichiometric in oxone and catalytic in fluorinated acetophenones. AB - [reaction: see text] Dioxiranes made in situ from the commercially available tetrafluoroacetophenones (7, 8) and pentafluoroacetophenone (9) are reported for highly efficient epoxidation of olefins for the first time. Studies showed that ketone 7, 8, or 9 can be used in catalytic amount (0.2 equiv) with only 0.6 equiv of Oxone (equal to 1.2 equiv of peroxymonosulfate) to selectively oxidize diene 1 to epoxide 2. The epoxidation reactions of dioxiranes of fluoroacetophenones are compared with the recently described complementary aliphatic acyclic fluorinated ketones. PMID- 12889892 TI - Weaker Lewis acid, better catalytic activity: dual mechanisms in perfluoroarylborane-catalyzed allylstannation reactions. AB - [reaction: see text] PhB(C(6)F(5))(2) exhibits much higher activity as a Lewis acid catalyst for the allylstannation of aromatic aldehydes than the stronger Lewis acid B(C(6)F(5))(3). This anomalous enhancement of catalytic activity for the weaker LA is shown to be partly due to decreased thermodynamic stability of ion pair 2b relative to 2a in the product-forming step of the reaction. A mechanistic path where the borane serves as the true LA catalyst is more important for the weakly Lewis acidic borane. PMID- 12889893 TI - Synthesis of N2 2'-deoxyguanosine adducts formed by 1-nitropyrene. AB - [reaction: see text] Synthesis of N(2) 2'-deoxyguanosine adducts formed by the ubiquitous carcinogen, 1-nitropyrene, is reported. Various conditions of Buchwald Hartwig palladium-catalyzed amination are examined. The most convenient synthetic approach involved a straightforward coupling between protected 2'-deoxyguanosine and bromonitropyrenes, which, upon reductive deprotection, provided excellent yield of the two 1-nitropyrene adducts. PMID- 12889894 TI - Facile synthesis of an azido-labeled thalidomide analogue. AB - [reaction: see text] A five-step synthesis of an azido-thalidomide analogue is presented. The sequence requires cheap and readily available starting materials and reagents, and only two steps require purification. Additionally, the azido labeled analogue possesses activity comparable to that of thalidomide in inhibiting the proliferation of human microvascular endothelial cells, thus providing impetus for its use as a potential photoaffinity label of thalidomide. PMID- 12889895 TI - Lanthanide triflate-promoted palladium-catalyzed cyclization of alkenyl beta-keto esters and amides. AB - [reaction: see text] Lanthanide triflates were found to promote the palladium catalyzed cyclization of alkenyl beta-keto esters and amides. In the presence of catalytic amounts of PdCl(2)(MeCN)(2) and Ln(OTf)(3), various alkenyl beta-keto esters and amides underwent regioselective cyclization reactions to give six-, seven-, or eight-membered-ring carbocycles in moderate to excellent yields. PMID- 12889896 TI - Polyfluorination of aryl alkyl sulfides by IF5 with concomitant migration of the arylthio group. AB - [reaction: see text] In the reaction of p-chlorophenyl alkyl sulfides with IF(5), polyfluorination reaction took place on the alkyl chain with the migration of the arylthio group. Consequently, p-chlorophenyl polyfluoroalkyl sulfides, having 3-7 fluorine atoms depending on alkyl chain length, could be obtained selectively. PMID- 12889897 TI - Decatungstate photocatalyzed oxidation of aryl alkanols. Electron transfer or hydrogen abstraction mechanism? AB - [reaction: see text] Decatungstate W(10)O(32)(4-) photosensitized oxidation of a series of para-X-substituted 1-aryl-1-alkanols was investigated. The only oxidation product of the side-chain of the 1-aryl-1-alkanol was the aryl ketone. The product analysis and kinetic data of the title reaction support a hydrogen atom transfer mechanism in the rate-determining step. PMID- 12889898 TI - Dioscorealides and dioscoreanone, novel cytotoxic naphthofuranoxepins, and 1,4 phenanthraquinone from Dioscorea membranacea Pierre. AB - [structure: see text] Commonly used among ingredients in Thai traditional anticancer preparations, the rhizome of Dioscorea membranacea Pierre was found potently cytotoxic and possibly contributed to such a therapeutic effect. Bioassay-guided isolation resulted in two novel cytotoxic naphthofuranoxepins, dioscorealides A (1) and B (2), and a new 1,4-phenanthraquinone, dioscoreanone (3). The structure determination, achieved mainly by means of NMR and CD spectral and X-ray crystallographic analyses, and cytotoxicity are discussed here. PMID- 12889899 TI - Tandem radical reactions and ring-closing metathesis. Application in the synthesis of cyclooctenes. AB - [reaction: see text] Fumarate- and acrylate-substituted oxazolidinones undergo tandem radical reaction to form dienes in moderate to good yields. The resulting dienes provide cyclooctenes in moderate to good yields after ring-closing metathesis (RCM). The role of the carbon backbone substituents and other variables in the efficiency of the eight-membered ring formation is discussed. PMID- 12889900 TI - Manganese(III)-catalyzed facile direct hydroperoxidation of some heterocyclic 1,3 dicarbonyl compounds. AB - [reaction: see text] The aerobic oxidation of 4-monosubstituted 1,2 diphenylpyrazolidine-3,5-diones 1 was carried out in the presence of a catalytic amount of manganese(III) acetate to quantitatively give the corresponding 4 hydroperoxypyrazolidinediones 2. A similar autoxidation of the 5-monosubstituted barbituric acids 5 and 3-butyl-4-hydroxy-2-quinolinone 7 also gave the corresponding hydroperoxides 5 and 8, respectively, in moderate to excellent yields. PMID- 12889901 TI - Simple and efficient protocol for the synthesis of functionalized styrenes from 1,2-dibromoethane and arylboronic acids. AB - [reaction: see text] A simple and efficient protocol for the preparation of functionalized styrenes is disclosed that employs the palladium-catalyzed cross coupling reaction of arylboronic acids with vinyl bromide, generated in situ from 1,2-dibromoethane. The reaction is carried out under mild reaction conditions. Compared with the cross-coupling reactions usually employed to obtain vinylarenes, this protocol is very simple, overcomes the inconvenience of using of ethylene under pressure, and uses air-stable and widely available arylboronic acids instead of vinyl organometallic reagents. PMID- 12889902 TI - Calyciphyllines A and B, two novel hexacyclic alkaloids from Daphniphyllum calycinum. AB - [structure: see text] Two types of Daphniphyllum alkaloids with unprecedented fused-hexacyclic ring systems, calyciphyllines A (1) and B (2), have been isolated from the leaves of Daphniphyllum calycinum (Daphniphyllaceae), and the structures and relative stereochemistry were elucidated on the basis of spectroscopic data. PMID- 12889904 TI - A stabilized formulation of IBX (SIBX) for safe oxidation reactions including a new oxidative demethylation of phenolic methyl aryl ethers. AB - [reaction: see text] SIBX is a nonexplosive formulation of IBX that can be used as a suspension in a variety of standard organic solvents such as refluxing EtOAc and THF to oxidize safely alcohols into aldehydes and ketones. The use of hot THF is limited to the oxidation of allylic and benzylic alcohols. Most yields are comparable to those obtained with IBX or DMP. SIBX can also be used to perform oxygenative demethylation of 2-methoxyarenols into orthoquinones and catechols. PMID- 12889903 TI - Intermolecular radical addition and addition/cyclization reactions of alkoxyamines onto nonactivated alkenes. AB - [reaction: see text] Alkoxyamines A, which are readily prepared from commercially available starting materials, undergo efficient thermal radical carboaminoxylations onto various nonactivated alkenes to provide 1,4 functionalized malonates B in good to excellent yields. The experiments are very easy to conduct. The carboaminoxylations can be combined with radical cyclization and fragmentation processes. PMID- 12889905 TI - beta-Nitro xanthates as olefin precursors. AB - [reaction: see text] Potassium O-ethyl xanthate readily adds to alpha,beta unsaturated nitro compounds to give stable beta-nitro xanthates, which undergo tin-free elimination to form olefins in good yield and good E selectivity upon simple heating with lauroyl peroxide in refluxing 1,2-dichloroethane. PMID- 12889906 TI - Synthesis of 2-aryl-oxazolo[4,5-c]quinoline-4(5H)-ones and 2-aryl-thiazolo[4,5 c]quinoline-4(5H)-ones. AB - [reaction: see text] Novel and highly efficient syntheses of oxazolo[4,5 c]quinoline-4(5H)-ones (1) and thiazolo[4,5-c]quinoline-4(5H)-ones (2) from ethyl 2-chlorooxazole-4-carboxylate (4) and ethyl 2-bromo-5-chlorothiazole-4 carboxylate (13), respectively, are described. PMID- 12889907 TI - Studies of intramolecular cyclizations of N-acyliminium ions derived from acyclic ketones: unanticipated stereochemical and structural results. AB - [reaction: see text] Intramolecular cyclizations of a series of (E)- and (Z) olefinic acyclic ketone-derived N-acyliminium ions have been studied. It has been found that both the course of the reaction and the stereochemistry of the products are critically dependent upon the tether length and olefin geometry of the cyclization substrate. PMID- 12889908 TI - Microwave-assisted solid-phase organic synthesis (MASPOS) as a key step for an indole library construction. AB - [reaction: see text] Microwave-assisted solid-phase organic synthesis (MASPOS) has been demonstrated to significantly facilitate the Cu(II)- or Pd(II)-mediated ring closure of the resin-bound 2-alkynylanilides. Under microwave irradiation at 200 degrees C [for Cu(OAc)(2), NMP] or 160 degrees C [for Pd(MeCN)(2)Cl(2), THF] for 10 min, 1-acyl-2-alkyl-5-arenesulfamoylindoles were obtained, after cleavage from the resin, in 95-99% purities and in 65-82% overall yields via a 5-step synthetic sequence. PMID- 12889909 TI - Unexpected stereochemistry in the lithium salt catalyzed ring expansion of nonracemic oxaspiropentanes. formal syntheses of (-)-(4R,5R)-muricatacin and the pheromone (R)-japonilure. AB - [reaction: see text] The stereochemistry of the cyclobutanones 3, obtained by lithium salt catalyzed ring expansion of the optically pure oxaspiropentanes 2, depends not only on the lithium salt but also on the stereochemistry of 2. They constitute the starting material for the syntheses of the acetogenin (-)-(4R,5R) muricatacin and the pheromone (R)-japonilure. PMID- 12889910 TI - Enantiospecific formal total synthesis of the tumor and GSK-3 beta inhibiting alkaloid, (-)-agelastatin A. AB - [reaction: see text] An enantiospecific total synthesis of Weinreb's advanced intermediate 2 for (-)-agelastatin A has been achieved from the Hough-Richardson aziridine 8. Noteworthy reactions in our sequence include the highly regioselective trans-diaxial ring-opening of 8 with azide ion to set up the vicinal diamido functionality present within (-)-2 and the Grubbs-Hoveyda ring closing metathesis (RCM) reaction that was used to construct its cyclopentene core. PMID- 12889911 TI - A novel and efficient total synthesis of cephalotaxine. AB - [reaction: see text] Total synthesis of cephalotaxine (CET), the parent member of a class of structurally unique antileukemia Cephalotaxus alkaloids, was accomplished on the basis of a conceptually novel strategy featuring transannular reductive skeletal rearrangements as the key transformations for the construction of the pentacyclic ring skeleton of CET. The synthetic potential of the designated Clemmensen-Clemo-Prelog-Leonard reductive rearrangement was demonstrated for the first time in a facile synthesis of the benzazepine subunit of CET. A novel endocyclic enamine (cyclopentenone) annulation was discovered and rationalized as an unusual azo-Nazarov-type cyclization. PMID- 12889912 TI - Novel solid-phase synthesis of 1,2-dialkoxyindoles. AB - [reaction: see text] A novel solid-phase synthesis of 1,2-dialkoxyindoles on SynPhase lanterns is described. A unique C-C bond formation involving a nucleophilic displacement of a solid-bound aryl fluorine by dimethyl malonate afforded the arylnitro methyl ester, which upon treatment with tin(II) chloride dihydrate gave the N-hydroxyindolone. Alkylation of the N-hydroxyindolones afforded the corresponding N-hydroxy-2-alkoxyindoles, which were further alkylated to give the 1,2-dialkoxyindole. A library of 64 (8R(1) x 8R(2)) discrete 1,2-dialkoxyindoles was prepared using a color encoding technique on SynPhase A-series lanterns. PMID- 12889913 TI - New strategy for the total synthesis of macrosphelides a and B based on ring closing metathesis. AB - [reaction: see text] A new total synthesis of macrosphelides A and B using ring closing metathesis (RCM) as a macrocyclization step is described. The substrate of the RCM could be synthesized from readily available chiral materials, methyl (S)-(+)-3-hydroxybutyrate and methyl (S)-(-)-lactate, with a high efficiency. The RCM proceeded in the presence of Grubbs' Ru-complex, providing a new effective synthetic route to these natural products. PMID- 12889914 TI - Reversal of diastereoselectivities in intra- and intermolecular reactions of 2 adamantanylidenes primarily caused by electron-donating and electron- withdrawing substituents on C5. AB - [reaction: see text] A reversal of diastereoselectivity was observed for novel 5 (trimethylsilyl)adamantan-2-ylidene (1c) with regard to 5-hydroxyadamantan-2 ylidene (1a). Ostensibly in intermolecular reactions, 5-substituted 2 adamantanylidenes (1) are sterically unbiased. However, inductive effects originating from the pendant group bend the divalent carbon bridge of 1 either toward (ERG's, e.g., -Si(CH(3))(3)) or away from (EWG's, e.g., -OH) the gamma position. Hence, the more exposed side is more susceptible to intermolecular reaction and the other side concomitantly undergoes intramolecular 1,3-CH insertions more readily. PMID- 12889915 TI - Origin of facial diastereoselection. Evidence for negative role of antiperiplanar hyperconjugation effects in the transition state of carbene insertion. AB - [reaction: see text] Quantitative analysis of the transition state (TS) structures of the insertion reactions of 5-X-2-adamantanylidenes (X = H, OH, NH(2), SiMe(3)) into MeOH or cyclohexane revealed that the magnitude of antiperiplanar hyperconjugative stabilization involving the incipient bond effects (the AP effect) decrease at TS, strongly suggesting that the AP effect contributes "net destabilization" of TS (i.e., increase in activation energy), in sharp contrast to the proposals of the Felkin-Anh and the Cieplak models. The equilibrium population between two carbene bridge-flipping proximal-distal conformers was found to be the origin of facial diastereoselection of these carbenes. PMID- 12889916 TI - Fmoc solid-phase synthesis of peptide thioesters by masking as trithioortho esters. AB - [reaction: see text] Total chemical synthesis of proteins by chemoselective ligation relies on C-terminal peptide thioesters as building blocks. Their preparation by standard Fmoc solid-phase peptide synthesis is made difficult by the lability of thioesters to aminolysis by the secondary amines used for removal of the Fmoc group. Here we present a novel backbone amide linker (BAL) strategy for their synthesis in which the thioester functionality is masked as a trithioortho ester throughout the synthesis. PMID- 12889917 TI - Regioselective formation, using orthogonal cysteine protection, of an alpha conotoxin dimer peptide containing four disulfide bonds. AB - [structure: see text] The combination of the cysteine thiol protecting groups Trt, Acm, tBu, and MeBzl were used for the regioselective formation of an alpha conotoxin dimer peptide containing four disulfide bridges. Additionally, a protocol is described whereby two one-pot oxidations were employed in order to improve the efficiency of the folding process. The target compound was produced in good yield. PMID- 12889918 TI - Solid-phase total synthesis of the pentacyclic system lamellarins U and L. AB - [reaction: see text] A total solid-phase synthesis of lamellarins U and L has been achieved. The conversion of an aldehyde group into a formate by a Baeyer Villiger reaction and a intramolecular [3 + 2] cycloaddition of a 3,4 dihydroisoquinolinium salt over a triple bond comprise the key steps of the process. Each transformation has been controlled with the proper spectroscopic and analytical methods. PMID- 12889919 TI - Photolysis of diacyl peroxides: a radical-based approach for the synthesis of functionalized amino acids. AB - [reaction: see text] Photolysis of the amino acid derived symmetrical and unsymmetrical diacyl peroxides at 254 nm at low temperature (-78 to -196 degrees C) generates various bis(amino acids) in a concise manner and with orthogonal protection. The methodology was applied to the synthesis of (4R)-5-propyl-L leucine (PrLeu), a component of HUN-7293. PMID- 12889920 TI - First asymmetric total synthesis of Us-7 and -8, novel D-seco Corynanthe-type oxindole alkaloids from Uncaria attenuata: structure revision of Us-7 and determination of absolute stereochemistry. AB - [reaction: see text] Starting from (S)-glycidol, the asymmetric total synthesis of novel D-seco Corynanthe-type oxindoles Us-7 and Us-8 was accomplished. The structure of Us-7 was revised from the reported structure 1 with the (3R,7S) form to structure 22 with (3S,7R), and the absolute stereochemistry at C15 of both alkaloids was established. PMID- 12889921 TI - Diethylphosphine oxide (DEPO): high-yielding and facile preparation of indolones in water. AB - [reaction: see text] Indolones are prepared in excellent yield at 80 degrees C in water by radical reaction (aryl radical formation, hydrogen atom abstraction, cyclization, and rearomatization) mediated by the reagent diethylphosphine oxide (DEPO). The reaction features V-501 as a water-soluble initiator; no other additives are needed. The process proceeds at a much lower temperature than is required for efficient reaction with toxic tributyltin hydride in benzene and permits significantly higher isolated yields than the corresponding reaction mediated by ethylpiperidine hypophosphite (EPHP). PMID- 12889922 TI - Photoinduced magnetization in a two-dimensional cobalt octacyanotungstate. AB - A new type of cyano-bridged Co-W bimetallic assembly, CsI[{CoII(3 cyanopyridine)2}{WV(CN)8}].H2O was synthesized. This compound exhibited a temperature-induced phase transition and a photoinduced magnetization. Irradiating with light induced a spontaneous magnetization with a magnetic phase transition temperature at 30 K. This photoinduced magnetization is due to the phase transition from CoIII(LS; S = 0)-WIV(S = 0) to CoII(HS; S = 3/2)-WV(S = 1/2) phases by the irradiation. PMID- 12889923 TI - Chain elongations by three carbon atoms at a time in the boron-catalyzed polymerization of methallyltriphenylarsonium ylide. AB - Methallyltriphenylarsonium ylide polymerized in the presence of catalytic amounts of trialkylboranes. Linear polymers containing a terminal alcohol function were obtained after alkaline hydrogen peroxide oxidation. The chain of these polymers is constituted of methyl-substituted double bonds, separated by one methylene group. Their structure is related to that of natural rubber, in which the double bonds are separated by two methylene groups. These polymers arise from successive chain elongations of three carbon atoms at a time. A cyclic, polymeric ketone was also prepared from a cyclic trialkylborane. The mechanism, which involves a boratropic rearrangement after every insertion of a monomer, is discussed. PMID- 12889925 TI - Bulky aluminum alkyl scavengers in olefin polymerization with group 4 catalysts. AB - The binding of H2O to MeAl(OAr)2 (1: Ar = 2,6-di-tert-butyl-4-methylphenyl) in THF-d8 at -40 degrees C provides aquo complex 2, the structure of which was determined by X-ray crystallography. Complex 2 is unstable above 0 degrees C in THF-d8 and decomposes to form ArOH (major), CH4 (minor), and a methyl aluminoxane of undetermined structure. Decomposition of 2 follows first-order kinetics with k = 3.0 x 10-4 s-1 at 5 degrees C. The hindered phenol ArOH slowly reacts with [Cp2ZrMe][MeB(C6F5)3] (4) in bromobenzene-d5 solution at 25 degrees C to furnish CH4 and [Cp2ZrOAr][MeB(C6F5)3] (5), the structure of which was confirmed by X-ray crystallography. This reaction follows second-order kinetics for [ArOH] = [4] = 0.045 M and with k = 2.8 x 10-3 M-1 s-1 at 25 degrees C. This corresponds to a rate that is >107 x slower than the apparent rate of ethylene insertion for 4 at 25 degrees C at typical concentrations encountered in olefin polymerization. The kinetic data, as well as control experiments involving the addition of ArOH to active catalyst producing poly(ethylene), demonstrate that ArOH has essentially no effect on polymerization kinetics involving 4. PMID- 12889924 TI - The first triple thiol-thiolate hydrogen bond versus triple diselenide bond that bridges two metal centers. AB - Treatment of fac(S)-[Rh(aet)3] (aet = 2-aminoethanethiolate) with aqueous HBF4 in air led to the protonation at coordinated thiolato groups to give a rhodium(III) dimer, [{Rh(aet)2(Haet)}{Rh(aet)(Haet)2}](BF4)3 ([1](BF4)3). On the other hand, similar treatment of fac(Se)-[Rh(aes)3] (aes = 2-aminoethaneselenolate) produced a dinuclear rhodium(III) complex, [Rh2(selenocystamine)3](BF4)6 ([2](BF4)6), because of the autoxidation of coordinated selenolato groups by air. The crystal structures of [1](BF4)3, DeltaDelta-[1](BF4)3, and [2](BF4)6 were determined by X ray crystallography. In [1]3+ two RhIII octahedrons are connected through a strong triple thiol-thiolate S-H...S hydrogen bond, while two RhIII octahedrons are directly joined by a triple diselenide bond in [2]6+. The cyclic voltammetry indicated that in acidic media the RhIII center in fac(Se)-[Rh(aes)3] is more easily oxidized to RhIV than that in fac(S)-[Rh(aet)3], which is responsible for the formation of coordinated diselenide bonds. PMID- 12889926 TI - An ionic liquid-supported ruthenium carbene complex: a robust and recyclable catalyst for ring-closing olefin metathesis in ionic liquids. AB - The synthesis of an ionic liquid-supported olefin metathesis catalyst derived from Grubb's ruthenium carbene complex is described. This new supported catalyst has been used in BMI.PF6 solvent, and this allowed success in solving the challenging problem of catalyst recycling. The IL catalyst in BMI.PF6 can be recovered and reused up to 10 consecutive cycles in RCM reactions of several dienes with excellent conversions. Moreover, the IL catalyst shows a remarkable stability in BMI.PF6 and can be stored several months without loss of activity. These results clearly demonstrate the importance of anchoring an imidazolium ionic liquid pattern to the catalyst to avoid its leaching from the BMI.PF6 phase. PMID- 12889927 TI - Tin-centered radical and cation: stable and free. AB - The first stable stannyl radical (tBu2MeSi)3Sn* (1) has been synthesized by the reaction of tBu2MeSiNa with SnCl2-dioxane in diethyl ether. The X-ray crystal structure and electron paramagnetic resonance (EPR) data of this radical show that 1 has a planar geometry, being a pi-radical in both the solid and the liquid states. One-electron oxidation of 1 with Ph3C+.B(C6F5)4- in benzene quantitatively produced the corresponding cation (tBu2MeSi)3Sn+.B(C6F5)4- (2), representing the stable free stannylium ion that has been fully characterized by X-ray analysis and NMR data. Being free, 2 features a record downfield shifted resonance for stannylium ions: +2653 ppm. PMID- 12889928 TI - Purely site-specific chemisorption and conformation of trimethylamine on Si(100)c(4 x 2). AB - One of the fundamental points of interest on the Si(100) surface is how the spatial localization of electron density on the buckled silicon dimer controls the site-specific reaction toward different Lewis acid and Lewis base molecules. We have investigated the adsorption of trimethylamine (TMA) on Si(100)c(4x2) using scanning tunneling microscopy (STM) at 80 K. The adsorbed TMA appears as a triangle-shaped bright protrusion in the occupied-state STM image. The triangle shaped protrusion is ascribed to three methyl groups in the adsorbed TMA. The center of the protrusion is located on the down atom site, which indicates that the adsorption of TMA occurs only on the down dimer atom. Thus, TMA adsorption on Si(100)c(4x2) is found to be purely site-specific on the down dimer atom and can be categorized in Lewis acid-base reaction. PMID- 12889929 TI - First examples of organophosphorus-containing materials for light-emitting diodes. AB - Exploiting the reactivity of the P-atom of phosphole-based oligomers, we have achieved access to the first organophosphorus-containing organic light-emitting diode (OLED) materials. The versatility of these P-materials is demonstrated with the synthesis of a corresponding gold complex that has also been used as an OLED material. Optimization of the OLED devices by doping the phosphole layer with a red fluorescent dye is described. PMID- 12889930 TI - Solventless polymerization: spatial migration of a catalyst to form polymeric thin films in microchannels. AB - This paper reports a simple, additive process to generate patterned polymer films without using any solvent. This process involves a highly efficient catalyst, a Grubbs's catalyst, and a volatile monomer, norbornene. The catalyst and monomers have higher local concentrations inside the microchannels, formed by contacting poly(dimethylsiloxane) stamps to a solid surface, and allow the polymeric thin films to be defined by the microchannels. The patterned thin film serves as an excellent resistant to reactive ion etching, which promises that this process is a complementary, useful alternative to spin-coating and plasma polymerization in microfabrication. PMID- 12889931 TI - Single-step in situ synthesis of polymer-grafted single-wall nanotube composites. AB - An in situ composite synthesis technique has been developed by grafting polystyrene chains onto single-wall carbon nanotubes (SWNTs) via a single-step debundling/polymerization scheme. The method, based on established anionic polymerization techniques, eliminates the need for nanotube pretreatment prior to functionalization and allows attachment of polymer molecules to pristine tubes without altering their original structure. The composites obtained contain well dispersed SWNTs with good nanotube-matrix interaction. The scheme is quite general in nature and can be applied to different polymer systems. The simplicity and scalability of the process can lead to the realization of superior nanotube based polymer composites for applications as advanced multifunctional structural materials. PMID- 12889932 TI - A cationic guest in a 24+ cationic host. AB - A new tetrahedral coordination cage M12L6 was prepared from a linear dipyrimidine ligand (L) and cis-protected palladium(II) (M). This cage showed unprecedented host-guest chemistry where the cationic host accommodated a cationic guest despite 24+ charges on the host framework. The unusual cation-cation host-guest chemistry is described by a unique onionlike shell structure of the host-guest complex where two cationic spheres are mediated by an anionic sphere of a counteranion assembly. PMID- 12889933 TI - Stereoelectronic effects on collagen stability: the dichotomy of 4-fluoroproline diastereomers. AB - Collagen is the most abundant protein in animals. Natural collagen consists of a triple helix of (Xaa-Yaa-Gly)n chains, in which the Xaa and Yaa residues are often l-proline. Here, a (2S,4S)-4-fluoroproline (flp) residue is shown to be greatly stabilizing in the Xaa position (but destabilizing in the Yaa position). In contrast, a (2S,4R)-4-fluoroproline (Flp) residue is shown to be greatly destabilizing in the Xaa position (but stabilizing in the Yaa position). The dichotomous effect of the diastereomers appears to arise from a gauche effect, which alters pyrrolidine ring pucker and hence properly (or improperly) preorganizes main-chain dihedral angles. Thus, the rational use of stereoelectronic effects can enhance the conformational stability of a protein. PMID- 12889934 TI - Molybdenum-phosphorus triple bond stabilization by ancillary alkoxide ligation: synthesis and structure of a terminal phosphide tris-1-methylcyclohexanoxide complex. AB - Using alcoholysis, we converted terminal phosphide PMo(N[i-Pr]Ar)3 into a new, monomeric terminal phosphide PMo(OR)3, where R = 1-methylcyclohexyl or 1 adamantyl. Dimerization of the PMo unit was observed upon alcoholysis with 2,6 dimethylphenol, and the dimer [PMo(N[i-Pr]Ar)(O-2,6-C6H3Me2)2]2 was isolated and characterized by X-ray crystallography. PMID- 12889935 TI - Synthesis, structure, and photoelectronic effects of a uranium-zinc-organic coordination polymer containing infinite metal oxide sheets. AB - The hydrothermal reaction of ZnUO2(OAc)4.7H2O with pyridine-2,3-dicarboxylic acid gives rise to a novel uranium-zinc-organic coordination polymer containing infinite U-O-Zn double sheets and organic ligands. Thermal and photoelectrochemical analyses indicate that the coordination polymeric compound not only has a high thermal stability but also exhibits interesting photoelectronic properties. PMID- 12889936 TI - [3+3]Cycloalkyne dimers linked by an azo group: a stable cis-azo compound forms polymeric aggregates by nonplanar pi-pi interactions. AB - We previously reported that the structure of the linker moiety controlled intramolecular and bimolecular aggregation of [3+3]cycloalkyne oligomers, which are cyclic acetylene derivatives containing helicenes. Here, novel [3+3]cycloalkyne dimers linked by the azo group are synthesized, and aggregation behaviors are studied, which turned out to be considerably different. The trans- and cis-azo compounds were synthesized by the oxidative coupling of a [3+3]cycloalkyne amine derivative, and the stereochemistry was determined by UV vis spectroscopy. 1H NMR, CD, gel permeation chromatography analysis, and vapor pressure osmometry in chloroform revealed that the trans-isomer forms a strong and selective bimolecular aggregate. The cis-isomer forms a trimolecular aggregate at a concentration below 1 mM and a polymeric aggregate at above 1 mM. Unlike known diaryl azo compounds, these azo isomers do not interconvert when subjected to heating or irradiation. In contrast, a model compound lacking the cyclic helicene structure isomerizes readily. PMID- 12889937 TI - Activation of alkyl halides via a silver-catalyzed carbene insertion process. AB - The silver complex [HB(3,5-(CF3)2Pz)3]Ag(THF) featuring a highly fluorinated tris(pyrazolyl)borate ligand catalyzes the formation of aliphatic carbon-halogen bond activation products under remarkably mild conditions. For example, the reaction between CHCl3 and ethyl diazoacetate (EDA) at room temperature in the presence of the silver catalyst afforded HClC(CO2Et)CCl2H in 60% yield. The presence of beta-hydrogens on the alkyl halide leads to net hydrogen halide addition to the carbene and an alkene. PMID- 12889938 TI - A cryptand/bisparaquat [3]pseudorotaxane by cooperative complexation. AB - The formation of the first cryptand/bisparaquat [3]pseudorotaxane appears to be highly cooperative. The apparent value of K2 is 17 times that of K1. PMID- 12889939 TI - TNA synthesis by DNA polymerases. AB - Threose nucleic acid (TNA), which has a repeat unit one atom shorter than that of DNA, is capable of Watson-Crick base pairing with DNA, RNA, and TNA. Because of its chemical simplicity, TNA is considered to be a possible progenitor of RNA. As an initial step toward developing the molecular tools necessary to investigate the functional capabilities of TNA by in vitro selection, we have screened a variety of DNA polymerases for TNA synthesis activity on a DNA template. We wish to report that several polymerases show surprisingly good ability to synthesize TNA using alpha-l-threofuranosyl thymidine-3'-triphosphate as a substrate. PMID- 12889940 TI - Unusual effects in the pd-catalyzed asymmetric allylic alkylations: synthesis of chiral chromans. AB - An examination of earlier reports of poor-to-modest results using Pd-catalyzed asymmetric allylic alkylations (AAA) to effect cyclization to form tetrasubstituted carbons reveals several novel factors that can influence this class of reactions. Thus, carboxylate has a major effect on such cyclizations wherein the ee increases from 14% ee favoring the S with no carboxylate to 84% ee favoring the R enantiomer in the presence of 1 equiv of carboxylate. Changing the double bond geometry from E to Z further increases the ee to 97%. Furthermore, the chiral catalyst that forms the R enantiomer with the E-alkene forms the S enantiomer with the Z alkene. In contrast to trisubstituted alkene substrates, disubstituted ones show a decrease in ee in going from the E to Z alkenes. The role of carboxylate appears to be a ligand to Pd during the catalytic cycle, a previously unsuspected phenomenon since such reactions are generally believed to involve pi-allylpalladium cationic complexes. The dependence upon alkene geometry helps define the nature of the chiral pocket which better accommodates a Z alkene compared to an E alkene. The results are compatible with the enantiodiscriminating step being ionization which occurs by coordination of the palladium to one of the two prochiral faces of the double bond. A synthesis of (+)-clusifoliol, a constituent of a folk medicine for treatment of malignant tumors, which also assigns the absolute configuration, illustrates the utility of the method. PMID- 12889941 TI - Stereospecific photochemical ring expansion of lithiated benzamides. AB - Treatment of N-benzyl benzamides with a strong base (LDA or t-BuLi) followed by irradiation with a 500 W tungsten lamp provides, according to the substitution pattern of the starting amides, either norcaradienes or cycloheptadienones by overall insertion of the N-benzyl group into the benzamide's aromatic ring system. Chiral benzamides undergo the ring expansion with high (sometimes complete) stereospecificity. The reaction appears to occur via a series of pericyclic reactions (photochemical or thermal sigmatropic rearrangements and thermal electrocyclic reactions) following an initial dearomatizing cyclization. PMID- 12889942 TI - Effect of catenation on protein folding stability. AB - Building on previous studies of the effects of covalent linking and backbone cyclization on protein folding stability, a theory is developed for the stabilization effect of catenating a dimeric protein. Relative to covalent linking, catenation is up to 1000-fold more powerful in stabilizing the folded structure. This dramatic stabilization derives from the dual effects of backbone cyclization and constrained relative motion between the subunits afforded by the catenane topology. PMID- 12889943 TI - Palladium-catalyzed [3+2] intramolecular cycloaddition of alk-5 ynylidenecyclopropanes: a rapid, practical approach to bicyclo[3.3.0]octenes. AB - Readily accessible hex-5-ynylidenecyclopropane derivatives cycloisomerize to bicyclic five-membered carbocycles upon heating with catalytic amounts of a palladium complex. PMID- 12889944 TI - Cucurbit[6]uril pseudorotaxanes: distinctive gas-phase dissociation and reactivity. AB - Cucurbit[6]uril forms a doubly charged complex with 1,4-butanediammonium cation that is observed using electrospray ionization Fourier transform mass spectrometry. Such 1:1 complexes are not observed for the smaller cucurbit[5]uril, which forms a 2:1 ammonium:cucurbituril complex instead. The 1:1 complex with cucurbit[6]uril is difficult to fragment via collisional activation; when it does fragment, both breakup of the cucurbituril cage and loss of the amine are observed. Further, the complex reacts with tert-butylamine via slow adduction. In contrast, nonrotaxane analogues (such as doubly charged 2:1 complexes of either protonated 1,4-butanediamine or protonated ethylenediamine with cucurbit[6]uril) fragment via easy loss of the intact amine upon collisional activation and react with tert-butylamine via rapid displacement of the original amine. On the basis of stoichiometry, fragmentation behavior, and reactivity, we conclude that the doubly charged complex of cucurbit[6]uril with 1,4 butanediammonium is a gas-phase pseudorotaxane. PMID- 12889945 TI - phi-Values beyond the ribosomally encoded amino acids: kinetic and thermodynamic consequences of incorporating trifluoromethyl amino acids in a globular protein. AB - The consequences of the substitution of 4,4,4-trifluorovaline for valine on the folding kinetics and thermodynamics of a globular protein are presented. Variants of the N-terminal domain of L9, a small alpha-beta protein, were prepared in which V3 or V21 was replaced by trifluorovaline. CD and NMR demonstrate that the structure is not perturbed. Both are more stable, the V3 variant by 0.8 kcal mol 1 and the V21 variant by 1.4 kcal mol-1. The increase of stability is significantly larger than that observed in coiled-coils on a per trifluoromethyl group basis. Folding is two-state, and the variants both fold faster than the wild type. The Phi-values are 0.16 and 0.11, respectively. PMID- 12889946 TI - Meta-conjugation and excited-state coupling in phenylacetylene dendrimers. AB - Traditional pictures of optical properties in phenylacetylene dendrimers view the molecule as a collection of independent chromophores, linked by meta-substitution at the central phenyl ring. While this picture is reasonable for explaining the observed absorption trends, it breaks down in describing the emission behavior. We utilize a combination of ab initio theory and experiment to demonstrate that differences in the absorbing and emitting states can be described using an exciton model with very weak chromophore coupling for the absorption geometry and strong coupling for the emission geometry. This result may have significant implications for the design of energy-funneling dendrimeric molecules. PMID- 12889947 TI - Mutasynthesis of enterocin and wailupemycin analogues. AB - Inactivation of the novel phenylalanine ammonia lyase gene encP, whose product is a key component in the biosynthetic pathway to benzoyl-coenzyme A (CoA) in the bacterium Streptomyces maritimus, resulted in the loss of production of the benzoate-primed polyketides enterocin and wailupemycin G. A series of cinnamate and benzoate derivatives were administered to the DeltaencP mutant, resulting in the formation of novel analogues bearing p-fluorobenzoate, 2- and 3 thiophenecarboxylate, and cyclohex-1-enecarboxylate residues. Given that the benzoate:CoA ligase EncN was evaluated to have broad in vitro substrate specificity towards aryl acids, the strict starter unit specificity observed in vivo indicates that the enterocin type II polyketide synthase (PKS) exerts selective control over the choice of starter units. This study represents the first mutasynthesis experiments with iterative type II PKSs. PMID- 12889949 TI - A new ruthenium-catalyzed cleavage of a carbon-carbon triple bond: efficient transformation of ethynyl alcohol into alkene and carbon monoxide. AB - We report a new and efficient ruthenium-catalyzed reaction that transforms ethynyl alcohol into alkene and carbon monoxide. The most efficient catalysts are TpRu(PPh3)(CH3CN)2PF6 (10 mol %) and lithium triflate (20 mol %). The mechanism of this reaction was elucidated using an isotope-labeling experiment. PMID- 12889948 TI - Studying the interaction of alpha-gal carbohydrate antigen and proteins by quartz crystal microbalance. AB - The interaction of alpha-galacosyl antigen and its binding protein (anti-Gal antibody) has been studied by quartz crystal microbalance through a SAM immobilization technique which overcomes the nonspecific interactions and molecule trapping for current piezoelectric biosensors. PMID- 12889950 TI - Design of base-discriminating fluorescent nucleoside and its application to t/c SNP typing. AB - We report a novel method for base detection using a base-discriminating fluorescent (BDF) nucleoside. We developed BDF probes containing methoxybenzodeazaadenine MDA and methoxybenzodeazainosine MDI, which give strong fluorescence only when the base on the complementary strand is cytosine and thymine, respectively. Thus, the MDA- and MDI-containing ODNs can be used as a very effective BDF probe for the detection of single base alterations, such as SNPs and point mutations. The present method using BDF probes is a very powerful tool for SNP typing that does not require any enzymes and time-consuming steps, and can avoid hybridization errors. In addition, a combination of MDA- and MDI containing BDF probes facilitates the T/C SNP typing of a heterozygous sample. PMID- 12889951 TI - Formation of nanostructured polymer filaments in nanochannels. AB - We describe the use of hard etching methods to create nanodimensional channels and their use as templates for the formation of polymer filament arrays with precise dimensional and orientational control in a single integrated step. The procedure is general as illustrated by the radical, coordination, and photochemical polymerizations that were performed in these nanochannels. The nanochannel templates (20 nm high, 20-200 nm wide, and 100 mum long) were fabricated by the combined use of electron-beam lithography and a sacrificial metal line etching technique. Radical polymerization of acrylates, metal catalyzed polymerization of norbornene, and photochemical polymerization of 1,4 diiodothiophene were carried out in these nanochannels. The polymers grown follow the dimensions and orientation of the channels, and the polymer filaments can be released without breaking. The approach opens up the possibility of just-in-place manufacturing and processing of patterns and devices from nanostructured polymers using well-established polymer chemistry. PMID- 12889952 TI - Addition of stable nitroxide radical to stable divalent compounds of heavier group 14 elements. AB - The reactions of stable cyclic dialkylgermylene 2 and dialkylstannylene 3 with 2,2,6,6-tetramethylpiperidinyl-1-oxy (TEMPO) radical (2 equiv) gave the corresponding 1:2 adducts 4 and 5, respectively, which were characterized by NMR, MS, and X-ray analyses. The kinetics of the stepwise addition of two TEMPO molecules to germylene 2 revealed that the initial addition of TEMPO to 2 was 1010 times slower than the second TEMPO addition to the resulting germyl radical. The origin of the rate difference was discussed on the basis of the qualitative perturbation theory. In contrast to the reactions of 2 and 3, the reaction of dialkylsilylene 1 with TEMPO gave an interesting 1,3-dioxadisiletane derivative. PMID- 12889953 TI - Diamond formation by reduction of carbon dioxide at low temperatures. AB - This Communication reports a low-temperature diamond synthesis technique, in which diamonds (10-250 mum) can form at a temperature as low as 440 degrees C by reduction of dense CO2 with metallic Na. The X-ray diffraction pattern of a powder sample shows three reflection peaks, indexed with 111, 220, and 311, corresponding unambiguously to cubic diamond. The Raman spectrum of the product exhibits an intense first-order peak at 1332 cm-1, which is the characteristic signature of the cubic diamond, indicating the formation of well-crystallized diamond. Carbon dioxide is a nontoxic low-energy molecule, abundant on earth. This novel reduction method could allow studies of large-size diamond growth using CO2 as the carbon source. PMID- 12889954 TI - Epicocconone, a novel fluorescent compound from the fungus epicoccumnigrum. AB - Epicocconone represents a new class of natural fluorescent probes based on a polyketide skeleton isolated from the fungus Epicoccum nigrum. Epicocconone is a small, cell permeable natural product with a high molar absorbtivity and a long Stokes' shift that will be useful in biotechnological applications. PMID- 12889955 TI - A new class of modular chiral ligands with fluxional groups. AB - In ligand design for asymmetric catalysis, the usual norm is to derive the face shielding elements from a chiral source. New ligands in which the face shielding is determined by fluxional groups are introduced. Their design, modular synthesis, and experiments to demonstrate the significance of the fluxional groups are discussed. The advantage is that the fluxional groups, introduced at a later stage, allow for simple tuning of the face shielding group. PMID- 12889957 TI - A stable silicon congener of highly strained bicyclo[3.2.0]hepta-1,3,6-triene. AB - A silicon-containing fused bicyclic compound with a highly strained bridgehead double bond, 2,3,6,7-tetra-tert-butyl-4-(tert-butyldimethylsilyl)-5-(tert butyldimethylsiloxy)-5-silabicyclo[3.2.0]hepta-1,3,6-triene (2), was synthesized quantitatively by the reaction of 1,2-bis-tert-butyl-4,4-bis(tert butyldimethylsilyl)-4-silatriafulvene (3) with di-tert-butylcyclopropenone (4) at 80 degrees C. An X-ray crystallographic analysis for 2 not only confirmed a bicyclic structure having a silacyclopentadiene (silole) ring fused with a silacyclobutene ring but also the remarkable deformation around the double bonds; the sum of the bond angles around the unsaturated bridgehead carbon was 333 degrees . The strain energy of a model 5-silabicyclo[3.2.0]hepta-1,3,6-triene was calculated at the MP2/6-31+G(d,p)//B3LYP/6-31+G(d) level (30.2 kcal/mol) to be comparable to that for parent bicyclo[3.2.0]hepta-1,3,6-triene (30.7 kcal/mol). Despite the high steric strain, 2 was stable enough to be kept intact for several months in the air. The high stability is ascribed to the effective steric protection of the ring system by the bulky substituents. PMID- 12889956 TI - 3-Boronoacrolein as an exceptional heterodiene in the highly enantio- and diastereoselective Cr(III)-catalyzed three-component [4+2]/allylboration. AB - This Communication reports the optimization of the first catalytic enantio- and diastereoselective hetero[4+2]/allylboration reaction to provide efficient access to alpha-hydroxyalkyl pyran derivatives. The key substrate 3-boronoacrolein pinacolate appears to be an exceptionally favorable heterodiene for use in Jacobsen's enantioselective reverse electron demand hetero[4+2] reaction with enol ethers, catalyzed by the tridentate (Schiff base)chromium complex 1. This one-pot three-component reaction was successfully applied to a concise total synthesis of (5R,6S)-6-acetoxy-5-hexadecanolide (2), the oviposition attractant pheromone of the female Culex mosquito capable of transmitting the West Nile virus. PMID- 12889958 TI - Effect of surface pressure on the insulator to metal transition of a langmuir polyaniline monolayer. AB - A remarkable change in the conductivity of a polyaniline (PAN) Langmuir monolayer in the conducting state, as a function of surface pressure, has been observed using scanning electrochemical microscopy (SECM). The film conductivity, as expressed by the SECM current response of a redox mediator, was measured in-situ in a Langmuir film balance. The conductivity of the film increases significantly with surface pressure, above a threshold value of ca. 20 mN m-1. PMID- 12889959 TI - Partitioning the loss in vancomycin binding affinity for D-Ala-D-Lac into lost H bond and repulsive lone pair contributions. AB - The binding affinity of 4, which incorporates a methylene (CH2) in place of the key linking amide of Ac2-l-Lys-d-Ala-d-Ala, for vancomycin was compared with that of Ac2-l-Lys-d-Ala-d-Ala (3) and Ac2-l-Lys-d-Ala-d-Lac (5). The vancomycin affinity for 4 was approximately 10-fold less than that of 3, but 100-fold greater than that of 5. This suggests that the reduced binding affinity of 5 (4.1 kcal/mol) may be attributed to both the loss of a key H-bond (1.5 kcal/mol) and a destabilizing lone pair/lone pair electrostatic interaction introduced with the ester oxygen of 5 (2.6 kcal/mol) with the latter, not the H-bond, being responsible for the largest share of the 1000-fold reduction. PMID- 12889960 TI - Inactivation of acetyl-CoA synthase/carbon monoxide dehydrogenase by copper. AB - Two recent crystal structures of acetyl-CoA synthase (ACS) from Moorella thermoacetica exhibited different metal contents and geometries at their active site, called the A-cluster. This led to the proposal of two catalytic mechanisms, one Ni-based, the other Cu-based. ACS was studied with respect to synthase activity, methyl group transfer activity, metal content, and EPR spectroscopy. Our results indicate that Cu is not required for catalysis and that it inactivates ACS by binding to the proximal site of the A-cluster. With Cu in this site, the A-cluster cannot accept a methyl group from the corrinoid-iron-sulfur protein, nor can it exhibit the NiFeC EPR signal after treatment with CO. PMID- 12889961 TI - A combined experimental and theoretical study of divalent metal ion selectivity and function in proteins: application to E. coli ribonuclease H1. AB - Structural and thermodynamic aspects of alkaline earth metal dication (Mg(2+), Ca(2+), Sr(2+), Ba(2+)) binding to E. coli ribonuclease H1 (RNase H1) have been investigated using both experimental and theoretical methods. The various metal binding modes of the enzyme were explored using classical molecular dynamics simulations, and relative binding free energies were subsequently evaluated by free energy simulations. The trends in the free energies of model systems based on the simulation structures were subsequently verified using a combination of density functional theory and continuum dielectric methods. The calculations provide a physical basis for the experimental results and suggest plausible role(s) for the metal cation and the catalytically important acidic residues in protein function. Magnesium ion indirectly activates water attack of the phosphorus atom by freeing one of the active site carboxylate residues, D70, to act as a general base through its four first-shell water molecules, which prevent D70 from binding directly to Mg(2+). Calcium ion, on the other hand, inhibits enzyme activity by preventing D70 from acting as a general base through bidentate interactions with both carboxylate oxygen atoms of D70. These additional interactions to D70, in addition to the D10 and E48 monodentate interactions found for Mg(2+), enable Ca(2+) to bind tighter than the other divalent ions. However, a bare Mg(2+) ion with two or less water molecules in the first shell could bind directly to the three active-site carboxylates, in particular D70, thus inhibiting enzymatic activity. The present analyses and results could be generalized to other members of the RNase H family that possess the same structural fold and show similar metal-binding site and Mg(2+)-dependent activity. PMID- 12889962 TI - Control of kinetics and thermodynamics of [1,5]-shifts by aromaticity: a view through the prism of Marcus theory. AB - The effects of aromatic stabilization on the rates of [1,5]-hydrogen shifts in a series of carbo- and heterocyclic dihydroaromatic compounds were estimated by B3LYP/6-31G computations. The aromatic stabilization energy of the product is directly translated into increased exothermicity of these reactions. Relative trends for a significant range of endothermic and exothermic [1,5]-shifts with different intrinsic activation energies are reliably described by Marcus theory. The effects of aromaticity or antiaromaticity are very large and can lead to dramatic acceleration or deceleration of [1,5]-hydrogen shifts and even to complete disappearance of the reaction barrier. Not only the activation energy but the shape and position of the reaction barrier can be efficiently controlled by changes in the aromaticity of the products, making these systems interesting models for studying hydrogen tunneling. Marcus theory can also be applied successfully to other pericyclic shifts such as [1,5]-shifts which involve chlorine and methyl transfer. PMID- 12889963 TI - Study of the chemistry of ortho- and para-biphenylnitrenes by laser flash photolysis and time-resolved IR experiments and by B3LYP and CASPT2 calculations. AB - The photochemistry of ortho-biphenyl azide (1a) has been studied by laser flash photolysis (LFP), with UV-vis and IR detection of the transient intermediates formed. LFP (266 nm) of 1a in glassy 3-methylpentane at 77 K releases singlet ortho-biphenylnitrene (1b) (lambda(max) = 410 nm, tau = 59 +/- 6 ns), which under these conditions decays cleanly to the lower energy triplet state. In fluid solution at 298 K, 1b rapidly (tau < 10 ns) partitions between formation of isocarbazole (4) (lambda(max) = 430 nm, tau = 70 ns) and benzazirine (1e) (lambda(max) = 305 nm, tau = 12 ns). Isocarbazole 4 undergoes a 1,5-hydrogen shift, with k(H)/k(D) = 3.4 at 298 K to form carbazole 9 and smaller amounts of two other isocarbazoles (7 and 8). Benzazirine 1e ring-opens reversibly to azacycloheptatetraene (1f), which serves as a reservoir for singlet nitrene 1b. Azacycloheptatetraene 1f ultimately forms carbazole 9 on the millisecond time scale by the pathway 1f --> 1e --> 1b --> 4 --> 9. The energies of the transient intermediates and of the transition structures connecting them were successfully predicted by CASPT2/6-31G calculations. The electronic and vibrational spectra of the intermediates, computed by density functional theory, support the assignment of the transient spectra, observed in the formation of 9 from 1a. PMID- 12889964 TI - Factors that determine the protein resistance of oligoether self-assembled monolayers --internal hydrophilicity, terminal hydrophilicity, and lateral packing density. AB - Protein resistance of oligoether self-assembled monolayers (SAMs) on gold and silver surfaces has been investigated systematically to elucidate structural factors that determine whether a SAM will be able to resist protein adsorption. Oligo(ethylene glycol) (OEG)-, oligo(propylene glycol)-, and oligo(trimethylene glycol)-terminated alkanethiols with different chain lengths and alkyl termination were synthesized as monolayer constituents. The packing density and chemical composition of the SAMs were examined by XPS spectroscopy; the terminal hydrophilicity was characterized by contact angle measurements. IRRAS spectroscopy gave information about the chain conformation of specific monolayers; the amount of adsorbed protein as compared to alkanethiol monolayers was determined by ellipsometry. We found several factors that in combination or by themselves suppress the protein resistance of oligoether monolayers. Monolayers with a hydrophobic interior, such as those containing oligo(propylene glycol), show no protein resistance. The lateral compression of oligo(ethylene glycol) monolayers on silver generates more highly ordered monolayers and may cause decreased protein resistance, but does not necessarily lead to an all-trans chain conformation of the OEG moieties. Water contact angles higher than 70 degrees on gold or 65 degrees on silver reduce full protein resistance. We conclude that both internal and terminal hydrophilicity favor the protein resistance of an oligoether monolayer. It is suggested that the penetration of water molecules in the interior of the SAM is a necessary prerequisite for protein resistance. We discuss and summarize the various factors which are critical for the functionality of "inert" organic films. PMID- 12889965 TI - First pseudorotaxane-like [3]complexes based on cryptands and paraquat: self assembly and crystal structures. AB - A new cryptand, bis(1,3,5-phenylene)tri(1,4,7,10-tetraoxadecyl) (3a), has been synthesized in good yield from bis(5-hydroxy-1,3-phenylene)-26-crown-8 (2a) and tri(ethylene glycol) ditosylate using pseudo-high dilution conditions. 3a forms a strong 1:1 complex with paraquat (1) in acetone solution with a high apparent association constant, 1.4 x 10(4) M(-)(1). A stoichiometry of 1:1 was also observed by mass spectrometry in the gaseous state. However, in the solid state, as determined by X-ray crystallography, the two complexes of 3a and the previously reported homologous cryptand, bis(1,3,5-phenylene)tri(1,4,7,10,13 pentaoxatridecyl) (3b), with paraquat (1) have 2:1 stoichiometry. A unique feature of these trimolecular pseudorotaxane-like complexes is that the guest occupies parts of the cavities of two cryptand molecules. For the first time it was found that in cryptand-based complexes, different stoichiometries are possible for the same host-guest pair. PMID- 12889966 TI - A heterocyclic peptide nanotube. AB - An open-ended hollow tubular structure is designed based on hydrogen-bond directed self-assembly of a chimeric cyclic peptide subunit comprised of alternating alpha- and epsilon-amino acids. The design features a novel 1,4 disubstituted-1,2,3-triazole epsilon-amino acid and its utility as a peptide backbone substitute. The N-Fmoc-protected epsilon-amino acid was synthesized in high yield and optical purity in three steps from readily available starting materials and was employed in solid-phase peptide synthesis to afford the desired cyclic peptide structure. The cyclic peptide self-assembly has been studied in solution by (1)H NMR and mass spectrometry and the resulting tubular ensemble characterized in the solid state by X-ray crystallography. PMID- 12889967 TI - Role of charge-transfer configurations on the addition reaction of aminoalkyl radicals onto acrylate double bonds. AB - The major factors affecting the reactivity of the important class of aminoalkyl radicals toward the addition reaction onto acrylate double bonds were studied. The rate constants of interaction, directly measured by using laser-induced photoacoustic calorimetry, range from 2.5 x 10(7) to 4.5 x 10(7) M(-)(1) s(-)(1). The reactivity of these radicals was compared to that of alkyl radicals. Calculations show that the reaction enthalpy is not the driving factor. Molecular orbital calculations of the transition states clearly demonstrated the large participation of charge-transfer configurations relative to nonpolar configurations, leading to a decrease of the barrier height. Moreover a quite good correlation is found when plotting the charge-transfer character in the transition state against the ionization potential. The roles of the different factors governing the reactivity of these species were elucidated. PMID- 12889968 TI - Resolution of the non-steady-state kinetics of the two-step mechanism for the Diels-Alder reaction between anthracene and tetracyanoethylene in acetonitrile. AB - The Diels-Alder reaction between anthracene and tetracyanoethylene in acetonitrile does not reach a steady-state during the first half-life. The reaction follows the reversible consecutive second-order mechanism accompanied by the formation of a kinetically significant intermediate. The experimental observations consistent with this mechanism include extent of reaction-time profiles which deviate markedly from those expected for the irreversible second order mechanism and initial pseudo first-order rate constants which differ significantly from those measured at longer times. It is concluded that the reaction intermediate giving rise to these deviations cannot be the charge transfer (CT) complex, which is formed during the time of mixing, but rather a more intimate complex with a geometry favorable to the formation of the Diels Alder adduct. The kinetics of the reaction were resolved into the microscopic rate constants for the individual steps. The rate constants, as shown in equation 1, at 293 K were observed to be 5.46 M(-)(1) s(-)(1) (k(f)), 14.8 s(-)(1) (k(b)), and 12.4 s(-)(1) (k(p)). Concentration profiles calculated under all conditions show that intermediate concentrations increase to maximum values early in the reaction and then continually decay during the first half-life. It is concluded that the charge-transfer complex may be an intermediate preceding the formation of the reactant complex, but due to its rapid formation and dissociation it is not detected by the kinetic measurements. PMID- 12889969 TI - "Belt and braces": a peptide-based linker system of de novo design. AB - A new self-assembling peptide-based linker is described. The system comprises three leucine-zipper sequences of de novo design: one peptide, "the belt", templates the co-assembly of the other two-half-sized peptides, "the braces". These basic features were confirmed by circular dichroism spectroscopy and analytical ultracentrifugation: when mixed, the three peptides reversibly formed a predominantly helical and stable 1:1:1 ternary complex. Surface plasmon resonance experiments demonstrated assembly of the complex on gold surfaces, while the ability of the system to bring together peptide-bound cargo was demonstrated using colloidal gold nanoparticles. In the latter experiments, the nanoparticles were derivatized with the brace peptides prior to the addition of the belt. Transmission electron microscopy images of the resulting networks revealed regular approximately 7 nm separations between adjacent particles, consistent with the 42-amino acid helical design of the belt and braces. To our knowledge, belt and braces is a novel concept in leucine-zipper assembly and the first example of employing peptides to guide nanoparticle assembly. PMID- 12889970 TI - The role of the HOOO(-) anion in the ozonation of alcohols: large differences in the gas-phase and in the solution-phase mechanism. AB - The mechanism of the ozonation of isopropyl alcohol was investigated for the gas and the solution phase using second-order many body perturbation theory and density functional theory (DFT) with the hybrid functional B3LYP and a 6 311++G(3df,3pd) basis set. A careful analysis of calculated energies (considering thermochemical corrections, solvation energies, BSSE corrections, the self interaction error of DFT, etc.) reveals that the gas-phase mechanism of the reaction is dominated by radical or biradical intermediates while the solution phase mechanism is characterized by hydride transfer and the formation of an intermediate ion pair that includes the HOOO(-) anion. The product distribution observed for the ozonation in acetone solution can be explained on the basis of the properties of the HOOO(-) anion. General conclusions for the ozonation of alcohols and the toxicity of ozone (inhaled or administered into the blood) can be drawn. PMID- 12889971 TI - Design of heterogeneous catalysts via multiple active site positioning in organic inorganic hybrid materials. AB - Catalytic materials bearing multiple sulfonic acid functional groups and positioned at varying distances from one another on the surface of mesoporous solids are prepared to explore the effects that the spatial arrangement of active sites have on catalytic activity and selectivity. A series of organosiloxane precursors containing either disulfide or sulfonate ester functionalities (synthons of the eventual sulfonic acid groups) are synthesized. From these molecular precursors, a variety of organic-inorganic hybrid, mesostructured SBA 15 silica materials are prepared using a postsynthetic grafting procedure that leads to disulfide and sulfonate ester modified silicas: [Si]CH(2)CH(2)CH(2)SS pyridyl, 2.SBA, [Si]CH(2)CH(2)CH(2)SSCH(2)CH(2)CH(2)[Si], 3.SBA, [Si]CH(2)CH(2)(C(6)H(4))(SO(2))OCH(2)CH(3), 4.SBA, and [Si]CH(2)CH(2)(C(6)H(4))(SO(2))OC(6)H(4)O(SO(2))(C(6)H(4))CH(2)CH(2)[Si], 6.SBA ([Si] = (tbd1;SiO)(x)()(RO)(3)(-)(x)()Si, where x = 1, 2). By subsequent chemical derivatization of the grafted species, thiol and sulfonic acid modified silicas are obtained. The materials are characterized by a variety of spectroscopic ((13)C and (29)Si CP MAS NMR, X-ray diffraction) and quantitative (TGA/DTA, elemental analysis, acid capacity titration) techniques. In all cases, the organic fragment of the precursor molecule is grafted onto the solid without measurable decomposition, and the precursors are, in general, attached to the surface of the mesoporous oxide by multiple siloxane bridges. The disulfide species 2.SBA and 3.SBA are reduced to the corresponding thiols 7.SBA and 8.SBA, respectively, and 4.SBA and 6.SBA are transformed to the aryl sulfonic acids 11.SBA and 12.SBA, respectively. 7.SBA and 8.SBA differ only in terms of the level of control of the spatial arrangement of the thiol groups. Both 7.SBA and 8.SBA are further modified by oxidation with hydrogen peroxide to produce the alkyl sulfonic acid modified materials 9.SBA and 10.SBA, respectively. The performances of the sulfonic acid containing SBA-15 silica materials (with the exception of 12.SBA) are tested as catalysts for the condensation reaction of phenol and acetone to bisphenol A. The alkyl sulfonic acid modified material 10.SBA derived from the cleavage and oxidation of the dipropyl disulfide modified material 3.SBA is more active than not only its monosite analogue 9.SBA, but also the presumably stronger acid aryl sulfonic acid material 11.SBA. It appears that a cooperative effect between two proximal functional groups may be operating in this reaction. PMID- 12889972 TI - Easily processable phenylene-thiophene-based organic field-effect transistors and solution-fabricated nonvolatile transistor memory elements. AB - The synthesis of a new series of mixed phenylene-thiophene oligomers is reported; 2,5-bis(4-n-hexylphenyl)thiophene (dH-PTP, 1), 5,5'-bis(4-n-hexylphenyl)-2,2' bithiophene (dH-PTTP, 2), 5,5' '-bis(4-n-hexylphenyl)-2,2':5',2' '-terthiophene (dH-PT(3)P, 3), 5,5' "-bis(4-n-hexylphenyl)-2,2':5',2' ':5' ',2' " quaterthiophene (dH-PT(4)P, 4), 1,4-bis[5-(4-n-hexylphenyl)-2-thienyl]benzene (dH PTPTP, 5), and 2,5-bis[4(4'-n-hexylphenyl)phenyl]thiophene (dH-PPTPP, 6) were characterized by (1)H NMR, elemental analysis, UV-visible spectroscopy, differential scanning calorimetry, and thermogravimetric analysis. Vacuum evaporated and solution-cast films were characterized by X-ray diffraction and scanning electron microscopy. All compounds display high p-type carrier mobilities as evaporated (up to 0.09 cm(2)/Vs) and as solution-cast (up to 0.03 cm(2)/Vs) films on both Si/SiO(2) and ITO/GR (glass resin) substrates. The straightforwardly synthesized dH-PTTP (2) displays an unprecedented combination of mobility, on/off ratio, stability, and processability. Both dH-PTTP (2) and dH PPTPP (6) display a reversible, tunable, and stable memory effect even as solution-cast devices, with turn-on characteristics shifting from accumulation mode to zero or depletion mode after a writing voltage V(w) is applied. The charge storage is distributed over the gate dielectric structure and is concentrated near the dielectric-semiconductor interface, as evidenced by the response of "floating gate" configuration devices. Simple nonvolatile elements have been fabricated by solution-only techniques on ITO substrates using spin coated glass resin, solution-cast oligomeric semiconductors, and painted graphite paste electrodes. PMID- 12889973 TI - Transition metal-catalyzed formation of boron-nitrogen bonds: catalytic dehydrocoupling of amine-borane adducts to form aminoboranes and borazines. AB - A mild, catalytic dehydrocoupling route to aminoboranes and borazine derivatives from either primary or secondary amine-borane adducts has been developed using late transition metal complexes as precatalysts. The adduct Me(2)NH.BH(3) thermally eliminates hydrogen at 130 degrees C in the condensed phase to afford [Me(2)N-BH(2)](2) (1). Evidence for an intermolecular process, rather than an intramolecular reaction to form Me(2)N=BH(2) as an intermediate, was forthcoming from "hot tube" experiments where no appreciable dehydrocoupling of gaseous Me(2)NH.BH(3) was detected in the range 150-450 degrees C. The dehydrocoupling of Me(2)NH.BH(3) was found to be catalyzed by 0.5 mol % [Rh(1,5-cod)(mu-Cl)](2) in solution at 25 degrees C to give 1 quantitatively after ca. 8 h. The rate of dehydrocoupling was significantly enhanced if the temperature was raised or if the catalyst loading was increased. The catalytic activity of various other transition metal complexes (Ir, Ru, Pd) for the dehydrocoupling of Me(2)NH.BH(3) was also demonstrated. This new catalytic method was extended to other secondary adducts RR'NH.BH(3) which afforded the dimeric species [(1,4-C(4)H(8))N-BH(2)](2) (2) and [PhCH(2)(Me)N-BH(2)](2) (3) or the monomeric aminoborane (i)Pr(2)N=BH(2) (4) under mild conditions. A new synthetic approach to the linear compounds R(2)NH-BH(2)-NR(2)-BH(3) (5: R = Me; 6: R = 1,4-C(4)H(8)) was developed and subsequent catalytic dehydrocoupling of these species yielded the cyclics 1 and 2. The species 5 and 6 are postulated to be intermediates in the formation of 1 and 2 directly from the catalytic dehydrocoupling of the adducts R(2)NH.BH(3). The catalytic dehydrocoupling of NH(3).BH(3), MeNH(2).BH(3), and PhNH(2).BH(3) at 45 degrees C to give the borazine derivatives [RN-BH](3) (10: R = H; 11: R = Me; 12: R = Ph) was demonstrated. TEM analysis of the contents of the reaction solution for the [Rh(1,5-cod)(mu-Cl)](2) catalyzed dehydrocoupling of Me(2)NH.BH(3) together with Hg poisoning experiments suggested a heterogeneous catalytic process involving Rh(0) colloids. PMID- 12889974 TI - Synthesis, solid-state crystal structure, and reactivity of a monomeric copper(I) anilido complex. AB - Synthesis and isolation of the Cu(I) amido complex (dtbpe)Cu(NHPh) (dtbpe = 1,2 bis(di-tert-butylphosphino)ethane) is accomplished upon reaction of [(dtbpe)Cu(mu Cl)](2) with LiNHPh. The anilido complex has been fully characterized by IR spectroscopy and multinuclear NMR spectroscopy as well as by single-crystal X-ray diffraction study. Salient features of the solid-state structure include an amido orientation that allows pi-interaction of the nitrogen-based lone pair with both the empty copper p-orbital and the pi-system of the phenyl substituent. A solid state X-ray diffraction study of [(dtbpe)Cu(NH(2)Ph)][BF(4)] has allowed a direct comparison of the structural features upon conversion of the amine ligand to an amido. The reactivity of the amido ligand of (dtbpe)Cu(NHPh) is consistent with nucleophilic character. For example, the formation of Ph(3)CNHPh is observed upon treatment with [Ph(3)C][BF(4)], and reaction at room temperature with EtX (X = Br or I) yields N-ethylaniline. The reactivity of (dtbpe)Cu(NHPh) is compared to that of the octahedral and d(6) complex TpRu(PMe(3))(2)(NHPh) (Tp = hydridotris(pyrazolyl)borate). PMID- 12889975 TI - Mechanisms of C-C and C-H alkane reductive eliminations from octahedral Pt(IV): reaction via five-coordinate intermediates or direct elimination? AB - The Pt(IV) complexes P(2)PtMe(3)R [P(2) = dppe (PPh(2)(CH(2))(2)PPh(2)), dppbz (o PPh(2)(C(6)H(4))PPh(2)); R = Me, H] undergo reductive elimination reactions to form carbon-carbon or carbon-hydrogen bonds. Mechanistic studies have been carried out for both C-C and C-H coupling reactions and the reductive elimination reactions to form ethane and methane are directly compared. For C-C reductive elimination, the evidence supports a mechanism of initial phosphine chelate opening followed by C-C coupling from the resulting five-coordinate intermediate. In contrast, mechanistic studies on C-H reductive elimination support an unusual pathway at Pt(IV) of direct coupling without preliminary ligand loss. The complexes fac- P(2)PtMe(3)R (P(2) = dppe, R = Me, H; P(2) = dppbz, R = Me) have been characterized crystallographically. The Pt(IV) hydrides, fac-P(2)PtMe(3)H (P(2) = dppe, dppbz), are rare examples of stable phosphine ligated Pt(IV) alkyl hydride complexes. PMID- 12889976 TI - Quantitative measure for the "nakedness" of fluoride ion sources. AB - A quantitative measure for the donor strength or "nakedness" of fluoride ion donors is presented. It is based on the free energy change associated with the transfer of a fluoride ion from the donor to a given acceptor molecule. Born Haber cycle calculations were used to calculate both the free energy and the enthalpy change for this process. The enthalpy change is given by the sum of the fluoride ion affinity of the acceptor (as defined in strict thermodynamic convention) and the lattice energy difference (DeltaU(POT)) between the fluoride ion donor and the salt formed with the acceptor. Because, for a given acceptor, the fluoride affinity has a constant value, the relative enthalpy (and also the corresponding free energy) changes are governed exclusively by the lattice energy differences. In this study, BF(3), PF(5), AsF(5), and SbF(5) were used as the acceptors, and the following seven fluoride ion donors were evaluated: CsF, N(CH(3))(4)F (TMAF), N-methylurotropinium fluoride (MUF), hexamethylguanidinium fluoride (HMGF), hexamethylpiperidinium fluoride (HMPF), N,N,N-trimethyl-1 adamantylammonium fluoride (TMAAF), and hexakis(dimethylamino)phosphazenium fluoride (HDMAPF). Smooth relationships between the enthalpy changes and the molar volumes of the donor cations were found which asymptotically approach constant values for infinitely large cations. Whereas CsF is a relatively poor F( ) donor [(U(POT)(CsF) - U(POT)(CsSbF(6))) = 213 kJ mol(-)(1)], when compared to N(CH(3))(4)F [(U(POT)(TMAF) - U(POT)(TMASbF(6))) = 69 kJ mol(-)(1)], a 4 times larger cation (phosphazenium salt) and an infinitely large cation are required to decrease DeltaU(POT) to 17 and 0 kJ mol(-)(1), respectively. These results clearly demonstrate that very little is gained by increasing the cation size past a certain level and that secondary factors, such as chemical and physical properties, become overriding considerations. PMID- 12889977 TI - Cationic hafnium silyl complexes and their enhanced reactivity in sigma-bond metathesis processes with Si-H and C-H bonds. AB - Reaction of the mixed-ring silyl methyl complex CpCp*Hf[Si(SiMe3)3]Me (4) with B(C6F5)3 in bromobenzene-d5 yielded the zwitterionic hafnium silyl complex [CpCpHfSi(SiMe3)3][MeB(C6F5)3] (7), which is stable for at least 12 h in solution. Addition of PhSiH3 to 7 rapidly produced HSi(SiMe3)3, CpCp*HfH(mu H)B(C6F5)3, and oligomeric silane products. Reactions of CpCp*Hf(SiR3)Me (SiR3 = SitBuPh2, SiHMes2) with B(C6F5)3 rapidly produced HSiR3 in quantitative yield along with unidentified hafnium-containing species. However, reactions of Cp2Hf(SiR3)Me (SiR3 = Si(SiMe3)3 (8), SitBuPh2 (9), SiPh3 (10)) with B(C6F5)3 quantitatively produced the corresponding cationic hafnium silyl complexes 12-14. The complex Cp2Hf(SitBuPh2)(mu-Me)B(C6F5)3 (13) was isolated by crystallization from toluene at -30 degrees C and fully characterized, and its spectroscopic properties and crystal structure are compared to those of its neutral precursor 9. The sigma-bond metathesis reaction of 13 with Mes2SiH2 yielded HSitBuPh2 and the reactive species Cp2Hf(eta(2)-SiHMes2)(mu-Me)B(C6F5)3 (16, benzene-d6), which was also generated by reaction of Cp2Hf(SiMes2H)Me (11) with B(C6F5)3. Spectroscopic data provide evidence for an unusual alpha-agostic Si-H interaction in 16. At room temperature, 16 reacts with benzene to form Cp2Hf(Ph)(mu Me)B(C6F5)3 (17), and with toluene to give isomers of Cp2Hf(C6H4Me)(mu Me)B(C6F5)3 (18-20) and Cp2Hf(CH2Ph)(mu-Me)B(C6F5)3 (21). The reaction with benzene is first order in both 16 and benzene. Kinetic data including activation parameters (deltaH = 19(1) kcal/mol; deltaS = -17(3) eu), a large primary isotope effect (kH/kD = 6.9(7)), and the experimentally determined rate law are consistent with a mechanism involving a concerted transition state for C-H bond activation. PMID- 12889978 TI - Cationic assembly of metal complex aggregates: structural diversity, solution stability, and magnetic properties. AB - The tetradentate imino-carboxylate ligand [L](2)(-) chelates the equatorial sites of Ni(II) to give the complex [Ni(L)(MeOH)(2)] in which a Ni(II) center is bound in an octahedral coordination environment with MeOH ligands occupying the axial sites. Lanthanide (Ln) and Group II metal ions (M) template the aggregation of six [Ni(L)] fragments into the octahedral cage aggregates (M[Ni(L)](6))(x)(+) (1: M = Sr(II); x = 2,2: M = Ba(II); x = 2, 3: M = La(III); x = 3, 4: M = Ce(III); x = 3, 5: M = Pr(III); x = 3, and 6: M = Nd(III); x = 3). In the presence of Group I cations, however, aggregates composed of the alkali metal-oxide cations template various cage compounds. Thus, Na(+) forms the trigonal bipyramidal [Na(5)O](3+) core within a tricapped trigonal prismatic [Ni(L)](9) aggregate to give ((Na(5)O) subset [Ni(L)](9)(MeOH)(3))(BF(4))(2).OH.CH(3)OH, 7. Li(+) and Na(+) together form a mixed Li(+)/Na(+) core comprising distorted trigonal bipyramidal [Na(3)Li(2)O](3+) within an approximately anti-square prismatic [Ni(L)](8) cage in ((Na(3)Li(2)O) subset [Ni(L)](8)(CH(3)OH)(1.3)(BF(4))(0.7))(BF(4))(2.3).(CH(3)OH)(2.75).(C(4)H(10)O)(0. ), 8, while in the presence of Li(+), a tetrahedral [Li(4)O](2+) core within a hexanuclear open cage [Ni(L)](6) in ((Li(4)O) subset [Ni(L)](6)(CH(3)OH)(3))2ClO(4).1.85CH(3)OH, 9, is produced. In the presence of H(2)O, the Cs(+) cation induces the aggregation of the [Ni(L)(H(2)O)(2)] monomer to give the cluster Cs(2)[Ni(L)(H(2)O)(2)](6).2I.4CH(3)OH.5.25H(2)O, 10. Analysis by electronic spectroscopy and mass spectrometry indicates that in solution the trend in stability follows the order 1-6 > 7 > 8 approximately 9. Magnetic susceptibility data indicate that there is net antiferromagnetic exchange between magnetic centers within the cages. PMID- 12889979 TI - alpha- and beta-A2Hg3M2S8 (A = K, Rb; M = Ge, Sn): polar quaternary chalcogenides with strong nonlinear optical response. AB - The closely related phases alpha- and beta-A(2)Hg(3)M(2)S(8) (A = K, Rb; M = Ge, Sn) have been discovered using the alkali polychalcogenide flux method and are described in detail. They present new structure types with a polar noncentrosymmetric crystallographic motif and strong nonlinear second-harmonic generation (SHG) properties. The alpha-allotropic form crystallizes in the orthorhombic space group Aba2 with a = 19.082(2) A, b = 9.551(1) A, c = 8.2871(8) A for the K(2)Hg(3)Ge(2)S(8) analogue, and a = 19.563(2) A, b = 9.853(1) A, c = 8.467(1) A for the K(2)Hg(3)Sn(2)S(8) analogue. The beta-form crystallizes in the monoclinic space group C2 with a = 9.5948(7) A, b = 8.3608(6) A, c = 9.6638(7) A, beta = 94.637 degrees for the K(2)Hg(3)Ge(2)S(8) analogue. The thermal stability and optical and spectroscopic properties of these compounds are reported along with detailed solubility and crystal growth studies of the alpha Kappa(2)Hg(3)Ge(2)S(8) in K(2)S(8) flux. These materials are wide gap semiconductors with band gaps at approximately 2.40 and approximately 2.64 eV for the Sn and Ge analogues, respectively. Below the band gap the materials exhibit a very wide transmission range to electromagnetic radiation up to approximately 14 microm. alpha-K(2)Hg(3)Ge(2)S(8) shows anisotropic thermal expansion coefficients. SHG measurements, performed with a direct phase-matched method, showed very high nonlinear coefficient d(eff) for beta-K(2)Hg(3)Ge(2)S(8) approaching 20 pm/V. Crystals of K(2)Hg(3)Ge(2)S(8) are robust to air exposure and have a high laser-damage threshold. PMID- 12889980 TI - Electronic excitation energies of Zn(i)O(i) clusters. AB - Time-dependent density-functional theory (TDDFT) is used to study the excitation energies of the global minima of small Zn(i)O(i) clusters, i = 1-15. The relativistic compact effective core potentials and shared-exponent basis set of Stevens, Krauss, Basch, and Jasien (SKBJ), systematically enlarged with extra functions, were used throughout this work. In general, the calculated excitations occur from the nonbonding p orbitals of oxygen. These orbitals are perpendicular to the molecular plane in the case of the rings and normal to the spheroid surface for 3D clusters. The calculated excitation energies are larger for ringlike clusters as compared to 3D clusters, with the excitation energies of the latter structures lying close to the visible spectrum. The difference between Kohn-Sham eigenvalues of the orbitals involved in the electronic excitations studied have also been compared to the TDDFT results of the corresponding excitations for two approximate density functionals, that is, MPW1PW91 and B3LYP, the latter being more accurate. Moreover, they approach the TDDFT value as the cluster size increases. Therefore, this might be a practical method for estimating excitation energies of large Zn(i)O(i) clusters. PMID- 12889981 TI - Structure and phase transitions of alkyl chains on mica. AB - We use molecular dynamics as a tool to understand the structure and phase transitions [Osman, M. A.; et al. J. Phys. Chem. B 2000, 104, 4433-4439. Osman, M. A.; et al. J. Phys. Chem. B 2002, 106, 653-662] in alkylammonium micas. The consistent force field 91 is extended for accurate simulation of mica and related minerals. We investigate mica sheets with 12 octadecyltrimethylammonium (C(18)) ions or 12 dioctadecyldimethylammonium (2C(18)) ions, respectively, as single and layered structures at different temperatures with periodicity in the xy plane by NVT dynamics. The alkylammonium ions reside preferably above the cavities in the mica surface with an aluminum-rich boundary. The nitrogen atoms are 380-390 pm away from the superficial silicon-aluminum plane. With increasing temperature, rearrangements of C(18) ions on the mica surface are found, while 2C(18) ions remain tethered due to geometric restraints. We present basal-plane spacings in the duplicate structures, tilt angles of the alkyl chains, and gauche-trans ratios to analyze the chain conformation. Agreement with experimental data, where available, is quantitative. In C(18)-mica with less than 100% alkali-ion exchange, the disordered C(18) rods in the island structures [Hayes, W. A.; Schwartz, D. K. Langmuir 1998, 14, 5913-5917] break at 40 degrees C. At 60 degrees C, the headgroups of the C(18) alkyl chains rearrange on the mica surface, and the broken chain backbones assume a coillike structure. The C(18) mica obtained on fast cooling of this phase is metastable due to slow reverse rearrangements of the headgroups. In 2C(18)-mica with 70-80% ion exchange, the alkali ions are interspersed between the alkyl chains, corresponding to a single phase on the surface. The observed phase transition at approximately 53 degrees C involves an increase of chain disorder (partial melting) of the 2C(18) ions without significant rearrangements on the mica surface. We propose a geometric parameter lambda for the saturation of the surface with alkyl chains, which determines the preferred self-assembly pattern, that is, islands, intermediate, or continuous. lambda allows the calculation of tilt angles in continuous layers on mica or other surfaces. The thermal decomposition seems to be a Hofmann elimination with mica as a base-template. PMID- 12889982 TI - Bonding and electronic structure in consanguineous and conjugal iron and rhenium sp carbon chain complexes [MC4M'](n)+: computational analyses of the effect of the metal. AB - Density functional theory has been used to probe the bonding and electronic properties of the homo- and heterobimetallic sp carbon chain complexes (ML(m), = (eta(5)-C(5)R(5))(eta(2)-R(2)PCH(2)CH(2)PR(2))Fe, (eta(5)-C(5)R(5))(NO)(PR(3))Re; z = 0-4). All neutral complexes are best described by MCtbd1;CCtbd1;CM electronic structures, in accord with much experimental data. The singlet dications are best described by cumulenic (+)M=C=C=C=C=M(+) valence formulations. However, the diiron and rhenium/iron dications are found to possess triplet states of nearly identical energy, clarifying experimental magnetic data. Their electronic structures have dominant *(+)MCtbd1;CCtbd1;CM(+)* character, with some spin delocalization onto the carbon chain. The mixed valence monocation radicals exhibit delocalized unpaired electrons, in accord with class III (strongly coupled) and II (weakly coupled) assignments made from experimental data earlier, with some spin density on the carbon chain. An isolable diiron trication has a doublet ground state, but some computational data suggest a close-lying quartet. For the unknown diiron tetracation, a bis(carbyne) or (2+)Fetbd1;CCtbd1;CCtbd1;Fe(2+) electronic structure is predicted. Calculated adiabatic ionization potentials show the iron endgroup to be more electron releasing than rhenium, in accord with electrochemical data. This polarizes the electronic structures of the rhenium/iron complexes. To help validate the computed model structures, crystal structures of ((eta(5)-C(5)Me(5))Fe(eta(2) dppe))(2)(mu-C(4)) and [((eta(5)-C(5)Me(5))Fe(eta(2)-dippe))(2)(mu-C(4))](3+) 3PF(6)(-) are determined. Data are analyzed with respect to related diruthenium and dimanganese complexes. PMID- 12889983 TI - On the nonpolar hydration free energy of proteins: surface area and continuum solvent models for the solute-solvent interaction energy. AB - Implicit solvent hydration free energy models are an important component of most modern computational methods aimed at protein structure prediction, binding affinity prediction, and modeling of conformational equilibria. The nonpolar component of the hydration free energy, consisting of a repulsive cavity term and an attractive van der Waals solute-solvent interaction term, is often modeled using estimators based on the solvent exposed solute surface area. In this paper, we analyze the accuracy of linear surface area models for predicting the van der Waals solute-solvent interaction energies of native and non-native protein conformations, peptides and small molecules, and the desolvation penalty of protein-protein and protein-ligand binding complexes. The target values are obtained from explicit solvent simulations and from a continuum solvent van der Waals interaction energy model. The results indicate that the standard surface area model, while useful on a coarse-grained scale, may not be accurate or transferable enough for high resolution modeling studies of protein folding and binding. The continuum model constructed in the course of this study provides one path for the development of a computationally efficient implicit solvent nonpolar hydration free energy estimator suitable for high-resolution structural and thermodynamic modeling of biological macromolecules. PMID- 12889984 TI - Influence of delocalization on the stability of dianions: study of a systematic series of dianions with growing electronic localization. AB - The electronic stability of a dianion is influenced by the degree of delocalization of its electrons, but it is generally not possible to separate this influence from other effects. Here, we investigate by theoretical means the sequence of dianions consisting of phen-1,4-ylenbis(ethynide) and seven of its derivatives obtained by hydrogenating the benzene core in several steps. These dianions are structurally similar and mainly differ by the degree of delocalization of their electrons. We present geometries and electron detachment energies computed at a correlated level of theory. The results point to a classification of the eight dianions in three distinct groups of electronic stability. We are able to explain this grouping by a simple resonance structure picture, which demonstrates why the dianions with more delocalized electrons are less stable. PMID- 12889986 TI - Impaired retrograde transmission of vasodilatory signals via the endothelium in pre-eclampsia: a cause of reduced tissue blood flow? AB - There is evidence that tissue blood flow is regulated by retrograde transmission of signals initiated at capillary and post-capillary sites, and transmitted via the endothelium to modulate pre-capillary resistance. We have used pre-eclampsia as a model to test the hypothesis that normal endothelium is required to enable adjustment of blood flow to match tissue requirements. Integrity of the endothelial pathway was assessed by measuring calf blood flow at increasing venous pressures, using an established small cumulative-step venous-congestion plethysmography protocol in ten women with pre-eclampsia, 17 normal pregnant controls and ten non-pregnant women. Endothelial cell activation was assessed by measuring plasma levels of the cell adhesion molecules, intercellular cell adhesion molecule-1 (ICAM-1), vascular cell-adhesion molecule-1 (VCAM-1) and E selectin. Baseline calf blood flow was significantly lower in pre-eclampsia than in the other two groups (P<0.0001; ANOVA). In the pre-eclampsia group, there was a fall in blood flow as venous congestion pressure was raised (P<0.0001; ANOVA). No such change was observed in the other two groups. A significant inverse correlation was observed between the reduction in blood flow in pre-eclampsia and the levels of E-selectin (r=-0.92, P=0.0002), VCAM-1 (r=-0.93, P=0.0008) and ICAM 1 (r=-0.86, P=0.001). The differences between the pre-eclamptic women and the other two groups support the notion that the failure to sustain blood flow during a cumulative pressure step protocol in the pre-eclamptic group might be influenced by interference with the retrograde transmission of signals via the endothelium in these patients. PMID- 12889987 TI - Isolation and characterization of haemoporin, an abundant haemolymph protein from Aplysia californica. AB - In the present study, we show the isolation and characterization of the protein haemoporin, which constitutes the second most abundant protein fraction in the haemolymph of the marine gastropod Aplysia californica. Although Aplysia is commonly used to investigate the molecular basis of learning, not much is known about the proteins in its haemolymph, which is in contact with the neurons owing to the open circulatory system of molluscs. In the native state, haemoporin is a macromolecular complex forming a cylinder with a central solvent-filled pore. The native complex most probably is a homopentamer made up from 70 kDa subunits with a molecular mass of 360 kDa and a sedimentation coefficient of 11.7 S. Prediction of the secondary structure by CD spectroscopy revealed that haemoporin contains 36% alpha-helices and 19% beta-strands. An absorption band in the 300-400 nm region indicates that haemoporin probably contains a bound substance. Haemoporin also contains a below average amount of tryptophan as evident from absorption and fluorescence spectra. The specific absorption coefficient at 280 nm (a (280 nm, 1 mg/ml)) varies between 0.42 and 0.59 l x g(-1) x cm(-1) depending on the method. The function of the protein is not yet known, but there are structural parallels between haemoporin and a pore protein reported previously in the haemolymph of another marine gastropod Megathura crenulata. The alanine-rich N-terminal sequence (AAVPEAAAEATAEAAPVSEF) is unique among protein sequences and indicates an alpha-helical structure. Whereas one side of the helix is hydrophobic and faces the interior of the protein, the other side contains a glutamic cluster, which may form the channel of the pore in the quaternary structure. Thus both proteins might belong to a new class of haemolymph proteins present in the haemolymph of marine gastropods. PMID- 12889988 TI - Moderate hyperalphalipoproteinaemia in a Brazilian population is related to lipoprotein lipase activity, apolipoprotein A-I concentration, age and body mass index. AB - We investigated 95 Brazilian adults, aged 21-79 years, who were divided into two groups defined as having high-density lipoprotein (HDL)-cholesterol concentrations above [hyperalphalipoproteinaemia (HALP); n=48] or below (controls; n=47) the 90th percentile of a local population. The activities of lipid transfer proteins and enzymes involved in the plasma reverse cholesterol transport and the prevalence of factors that modulate HDL metabolism (alcohol consumption, ponderosity, physical exercise, menopause and use of hormone replacement treatment in women and smoking) were measured, as well as the prevalence of cardiovascular disease and of its various risk factors. The two groups showed no differences in their frequencies of cardiovascular disease. The HDL2/HDL3-cholesterol and triacylglycerol (triglyceride) ratios and the activities of the phospholipid transfer protein (PLTP) and cholesteryl ester transfer protein (CETP) were similar in both groups. Lipoprotein lipase (LPL) and hepatic lipase (HL) activities were 35% higher (P=0.0002) and 40% lower (P=0.0006) respectively, in HALP compared with control subjects. In a multivariate analysis, HDL-cholesterol and its subfractions were influenced by LPL, apolipoprotein A-I, age (negative relationship) and body mass index (negative relationship). Use of alcohol and ponderosity, as well as the interaction of these factors, explained the LPL activity. HL activity was modulated by smoking, and hormone-replacement therapy influenced the apolipoprotein A-I concentration. CETP activity was influenced by race and PLTP by age. The unique phenotype found in this Brazilian HALP population, namely low HL and high LPL activities, could be determined mostly by genetic components, on which future work will focus. PMID- 12889989 TI - Short- and long-term reproducibility of autonomic measures in supine and standing positions. AB - Autonomic nervous tests and heart rate variability (HRV) have been used to assess cardiac autonomic function and to evaluate long-term prognosis. The aim of this study was to evaluate the short- and long-term reproducibility of HRV parameters and autonomic nervous tests according to body position (supine or standing). The study group consisted of 26 healthy subjects. Autonomic nervous tests and HRV were performed twice during the day and the results were averaged. The protocol was then repeated 3 days after each examination and also after 6 and 24 months. Autonomic nervous tests included deep breathing, Valsalva manoeuvre and isometric muscle exercise (handgrip), as well as blood pressure and heart rate in response to standing. ECG recordings were taken for 10 min during spontaneous breathing for HRV analysis. We found that the reproducibility of some parameters of the autonomic nervous test were independent of body position [E/I ratio (heart rate response to deep breathing)], whereas other parameters were dependent on body position (Valsalva manoeuvre and blood pressure response to sustained handgrip). In addition, within-day measurements of those parameters varied from non reproducible (Valsalva ratio, handgrip and blood pressure response to standing) to moderately reproducible [E/I ratio and 30/15 ratio (heart rate response to standing)]. Among the HRV parameters, we found that total power (TP), low (LF)- and high (HF)-frequency were reproducible not only for measurements made within the same day, but also during short- and long-term observations, and only the LF/HF ratio was dependent on body position. We conclude that only a few autonomic nervous tests are reproducible in the short- and long-term. Because HRV parameters obtained during spontaneous respiration showed high reproducibility for measurements made within the same day as well as in the short- and long-term, they should be used instead of autonomic nervous tests when long-term observations are carried out in a healthy population. PMID- 12889990 TI - Current and potential uses for DNA microarrays in transplantation medicine: lessons from other disciplines. AB - DNA microarrays are used to study simultaneous gene expression in thousands of genes. This tool has moved beyond proof-of-principle and its integration into medical practice is slowly becoming a reality. This technology has enabled unparalleled progress into the study of complex polygenic diseases. Although cancer research introduced DNA microarrays into the medical arena other disciplines are beginning to exploit the power of this technology to advance medical research. In this review we outline aspects of the design of a microarray experiment from the choice of platform, through the experimental procedure to the analysis of the results. We review the current applications and speculate on potential applications of this technology with particular reference to transplantation medicine. PMID- 12889991 TI - Evidence of a limited contribution of feto-maternal interactions to trophoblast differentiation along the invasive pathway. AB - Trophoblast differentiation is a key event in human placental development. During extravillous trophoblast (EVT) differentiation, stem cells from the anchoring villi detach from their basement membrane and proliferate to form aggregates called trophoblast cell columns (TCCs). They subsequently invade the decidua and differentiate into interstitial and endovascular trophoblasts. The influence of the decidua on EVT differentiation is controversial. We therefore compared the pattern of trophoblast differentiation marker expression in viable intrauterine and tubal pregnancies, as decidual cell markers (prolactin [PRL] and insulin-like growth factor binding Protein-1 [IGFBP1]) were only expressed in endometrial implantation sites. Extravillous trophoblast differentiation in anchoring villi from uterine and ectopic pregnancies exhibited a comparable phenotypical switch: alpha6 integrin subunit, E-cadherin, EGF receptor, Ki 67 and connexin 40 were localized in the proximal part of the TCC, while alpha5beta1 and alpha1 integrins, c-erb B2, hPL and HLA-G were expressed by invasive cytotrophoblasts. The cyclin-dependent kinase inhibitors p16 and p57 were mainly detected in invasive cytotrophoblasts some distance from the columns. However, the TCC was markedly longer in tubal pregnancy than in intrauterine pregnancy. These findings suggest that the decidua is not necessary to trigger EVT invasion, but that it is likely to limit the extent of the TCC and to accelerate the onset of EVT migration. PMID- 12889992 TI - Loss of Ly-6A.2 expression on immature developing T cells in the thymus is necessary for their normal growth and generation of the Vbeta T-cell repertoire. AB - Stage-specific expression of a number of cell-surface and signaling proteins is critical for normal development of T cells in the thymus. Equally important may be the loss of expression/signaling of developmentally regulated proteins for proper transitioning of developing T cells into thymic subsets. Ly-6A.2 exhibits a regulated pattern of expression on T cells maturing in the thymus, and dysregulating its expression results in arrest of developing T cells within the CD3-CD4-CD8- triple negative (TN) stage where the normal expression of Ly-6A.2 is extinguished. To further characterize the mechanisms underlying this block, we examined whether cell signaling and/or cell adhesion properties of the Ly-6A.2 molecule influenced the block in T-cell development. Analysis of bone marrow chimeras generated by injecting CFSE-labeled Ly-6A.2 transgenic bone marrow cells into irradiated syngeneic non-transgenic mice revealed normal trafficking of developing T cells from the cortex into the medulla. Production of LAT but not p56lck was diminished in CD4-CD8- DN cells from Ly-6A.2 dysregulated mice when compared with control littermates. Dysregulated expression of Ly-6A.2 did not suppress endogenous TCR-Vbeta expression. Finally, dysregulated expression of Ly 6A.2 enhanced apoptosis of an immature CD4+CD8+ (DP) subset of developing cells and altered the selected TCR-Vbeta repertoire. Taken together, these observations indicate that the termination of Ly-6A.2 expression and signaling within the CD4 CD8-CD3- subset of developing T cells is an important checkpoint during normal thymic development. PMID- 12889993 TI - Associations of HLA microsatellites with rheumatoid arthritis in Singaporean Chinese. AB - Rheumatoid arthritis in Singaporean Chinese has previously been shown to be associated with the DRB1*0405, DRB1*1001 haplotypes and to the DRB1*0901 haplotype when the former two were removed. The present paper focused on eight HLA associated microsatellite markers (TNFa, TNFd, D6S273, TAP1CA, DQCAR, DQCARII, D6S2222, D6S2223) and their allelic associations with Chinese RA. 60 RA patients and 75 healthy controls were studied. It appeared that DQCARII*194/DRB1*0405/TNFa*117 was part of the extended haplotype predisposed to RA, whereas DRB1*0901/D6S273*128 contributed to susceptibility to RA to a lesser degree in Singaporean Chinese. Additionally, a negative association with DQCAR*186/DRB1*0301/D6S273*122/TNFd*124 was observed. No association with disease development was observed in this study. PMID- 12889994 TI - Specificity and functional characteristics of anti-HLA-A mAbs LGIII-147.4.1 and LGIII-220.6.2. AB - Only three anti-HLA-A monoclonal antibodies (mAbs) have been described in the literature. Two of them recognize determinants shared by only a few HLA-A allospecificities. The third one, mAb 3G11, recognizes a determinant shared by most HLA-A allospecificities. Being an IgM, the latter mAb is not likely to be a useful probe in immunohistochemical reactions and in functional assays. Therefore, in the present study we have characterized the specificity of the mAbs LGIII-147.4.1 and LGIII-220.6.2. The two mAbs that do not share idiotypic determinants recognize distinct but spatially close antigenic determinants expressed on most of the gene products of the HLA-A locus. Specifically, the determinant recognized by mAb LGIII-220.6.2 is expressed on HLA-A1, -A2, -A3, A26, -A28, -A29, -A30, -A33, -A36, -A74 and -A80 allospecificities. The determinant recognized by mAb LGIII-147.4.1, which appears to be located on the amino-acid residues 79-83 of the heavy chain, is expressed on all HLA-A allospecificities but HLA-A23, -A24, -A25 and -A32. Because of its broad reactivity, the mAb LGIII-147.4.1 was characterized in a number of assays. It was found to be a useful probe to measure the HLA-A antigen level in serum, to assess the HLA-A restriction of cytotoxic T lymphocytes (CTL) and to monitor HLA-A antigen expression in normal and malignant lesions. PMID- 12889995 TI - Expression and function of HLA-DR3 and DQ8 in transgenic mice lacking functional H2-M. AB - H2-M or HLA-DM are non-classical class II molecules encoded by the MHC and play an important role during antigen presentation. They catalyze exchange of CLIP (Class II-associated invariant chain peptide) or other low-affinity peptides bound to class II molecules for peptides capable of more efficient binding. The phenotype of mice lacking H2-M is determined by the allotype of the MHC class II molecules expressed. In general, H2-M deficiency does not affect the surface expression of mature class II molecules. The class II molecules in such cases predominantly contain CLIP in their peptide-binding groove. In some mice strains, H2-M deficiency results in defective CD4+ T-cell development accompanied by defective responses to conventional antigens and superantigens. Even though the HLA class II molecules show similar dependency for HLA-DM for presenting antigens in vitro, their interaction in vivo is not known. By using transgenic approach we show here that DQ8 and DR3 are expressed at normal levels in H2-M-deficient mice and the CD4+ T-cell development is unaltered. However, the ability of DQ8 molecules to present peptide antigens is compromised in a H2-M-deficient state. Presentation of exogenous bacterial superantigens by both DQ8 and DR3 is unaffected in H2-M-deficient mice. Unexpectedly, Staphylococcal Enterotoxin B induced systemic IFN-gamma production was significantly higher in H2-M-deficient DQ8/DR3 transgenic mice and these mice were susceptible to SEB-induced toxic shock at doses that are non-lethal to H2-M-sufficient counterparts. PMID- 12889996 TI - HLA DR-DQ-encoded genetic determinants of childhood-onset type 1 diabetes in Finland: an analysis of 622 nuclear families. AB - The diabetes predisposing effect of HLA genes is defined by a complex interaction of various haplotypes. We analyzed the disease association of HLA DRB1-DQA1-DQB1 genotypes in a large nuclear family cohort (n = 622) collected in Finland. Using the affected family based artificial control approach we aimed at characterizing all detectable disease-specific HLA haplotype and genotype effects. The DRB1*0401 DQB1*0302 haplotype was the most prevalent disease susceptibility haplotype in the Finnish population followed by (DR3)-DQA1*05-DQB1*02 and DRB1*0404-DQB1*0302. DRB1*0405-DQB1*0302 conferred the highest disease risk, although this haplotype was very rare. The DRB1*04-DQB1*0304 was also associated with increased disease risk, an effect detected for the first time in the Finnish population. The following haplotypes showed significant protection from the disease and are listed in decreasing order of the strength of their effect: (DR7)-DQA1*0201 DQB1*0303, (DR14)-DQB1*0503, (DR15)-DQB1*0602, DRB1*0403-DQB1*0302, (DR13) DQB1*0603, (DR11/12/13)-DQA1*05-DQB1*0301, (DR1)-DQB1*0501. In addition to the DRB1*0401/0404-DQB1*0302/(DR3)-DQA1*05-DQB1*02 genotype and DRB1*04-DQB1*0302 homozygous genotypes, heterozygous combinations DRB1*0401-DQB1*0302/(DR13) DQB1*0604, approximately /(DR8)-DQB1*04, approximately /(DR9)-DQA1*03-DQB1*0303, approximately /(DR1)-DQB1*0501 and approximately /(DR7)-DQA1*0201-DQB1*02 were also disease-associated. As a new finding in this population, the (DR3)-DQA1*05 DQB1*02 homozygous and (DR3)-DQA1*05-DQB1*02/(DR9)-DQA1*03-DQB1*0303 heterozygous genotypes conferred disease susceptibility. Similarly, the DRB1*0401 DQB1*0302/(DR13)-DQB1*0603 genotype was disease predisposing, implying that DQB*0603-mediated protection from diabetes is not always dominant. Comparison of our findings with published data from other populations indicates a significant disease-specific heterogeneity of the (DR8)-DQB1*04, (DR7)-DQA1*0201-DQB1*02 and (DR3)-DQA1*05-DQB1*02 haplotypes. PMID- 12889997 TI - Association between polymorphisms in the human chemokine receptor genes CCR2 and CX3CR1 and rheumatoid arthritis. AB - Cell trafficking into the rheumatoid synovium is thought to play an important role in the inflammation seen in rheumatoid arthritis. Chemokine receptors play a central role in this process, and several common variants are known, including the CCR2 variant, CCR2-64I, and two variants of the CX3CR1 gene, V249I and T280M. All three variants result in functional amino acid substitutions. We studied the association of these chemokine receptor variants with susceptibility to and severity of rheumatoid arthritis in two Dutch patient populations; 282 consecutive rheumatoid arthritis patients from a rheumatology outpatient clinic, and a cohort of 101 female rheumatoid arthritis patients, followed closely for a 12-year period, from whom hand and feet X-rays taken at three year intervals were scored and analyzed in this study. Although there was a trend towards increased severity of disease in patients carrying CX3CR1 variants, this was not independent of known risk factors. We found no evidence for a significant independent role for the CCR2 and CX3CR1 variants in the susceptibility to or severity of rheumatoid arthritis. PMID- 12889998 TI - Novel HLA B*2714 and B*2708 allele associations in seronegative spondarthritis patients and haemophilia patients with chronic synovitis in India. AB - HLA B27 association with seronegative spondarthritis (SSA) has been recognized for over a decade and its association with chronic synovitis in Haemophilia patients has been recently reported. We characterized the HLA B27 subtypes in a cohort of 98 B27 positive individuals with SSA (n = 45), haemophilia with chronic synovitis (n = 21) and normal controls (n = 32), using polymerase chain reaction based sequence specific primers (PCR-SSP) and reverse line strip (RLS) techniques. The results revealed a significant association for a novel allele B*2714 in SSA along with the previously reported B*2705 allele. We also found a significant association between haemophilia patients with chronic synovitis from India and a hitherto unidentified allele, B*2708. PMID- 12889999 TI - Novel HLA-Cw*03 allele, Cw*030303, identified by nucleotide sequencing. AB - In this paper we report the identification of a new HLA- Cw*03 allele in a sample that has been distributed in the International UCLA Terasaki Cell/DNA Exchange. This novel allele officially designed Cw*03033 and renumbered to Cw*030303 (2) found in cell no. 1054 from an Asian Indian donor differs from Cw*030301 by a silent substitution at codon 128, GGG-->GGA (nucleotide position 384). This new allelic variant was confirmed by several other laboratories participating in the UCLA Terasaki Cell/DNA Exchange (3). PMID- 12890000 TI - High resolution sequence-based DPB1 typing identified two novel DPB1 alleles, DPB1*9401 and DPB1*9501, from a Kenyan population. AB - Two novel DPB1 alleles, DPB1*9401 and DPB1*9501, were identified from a Kenyan population during sequence-based HLA-DPB1 typing. Molecular cloning and sequencing of multiple clones confirmed that one of the new DPB1 alleles is identical to DPB1*0402 at exon 2 except for a single nucleotide substitution (CGG -->TGG), changing codon 70 from Arg to Trp. The new allele has been named DPB1*9401. This is the first report of polymorphism at codon 70 of HLA-DPB1 alleles. New codon combinations have been identified in another novel DPB1 allele named DPB1*9501. The extensive diversity at DPB1 locus of this East African population is being revealed by high resolution sequence-based DPB1 typing. PMID- 12890001 TI - Sequence-based typing identifies a novel HLA-DPB1 allele, DPB1*9601. AB - In this report we describe the identification of a novel HLA-DPB1 allele, DPB1*9601, found in a Caucasian individual sample named ucla#356. The new allele was detected in the DNA of ucla#356 during routine HLA sequence-based typing (SBT) of samples participating in the UCLA International HLA DNA Exchange (number 55) for HLA DNA Proficiency Testing. DPB1*9601 was identical to DPB1*3901 except for a single nucleotide substitution 'G'-->'C' in previously constant position 277 (position 177, respectively, counting only exon 2). This nucleotide change causes an amino acid substitution from aspartic acid in DPB1*3901 to histidine at codon 64 in the novel allele. This new allele has been submitted to the EMBL database and has been assigned the accession number AJ514871. The WHO Nomenclature Committee has officially assigned the name DPB1*9601. PMID- 12890002 TI - Anemia in children after transplantation: etiology and the effect of immunosuppressive therapy on erythropoiesis. AB - Anemia in children after renal transplantation is more common than previously appreciated. Multiple factors appear to play roles in the development of post transplant anemia, the most common of which is absolute and/or functional iron deficiency anemia. Most experts recommend that iron limited anemias in transplant patients should be diagnosed using the same criteria as for chronic renal failure patients. Serum erythropoietin (EPO) levels are expected to normalize after a successful renal transplantation with a normal kidney function, yet both EPO deficiency and resistance have been reported. While no large controlled trials comparing the effect of different immunosuppressive agents on erythropoiesis after transplantation have been performed, generalized bone marrow suppression attributable to azathioprine (AZA), mycophenolate mofetil (MMF), tacrolimus, antithymocyte preparations has been reported. Pure red cell aplasia (PRCA) occurs rarely after transplantation and is characterized by the selective suppression of erythroid cells in the bone marrow. PRCA has been reported with the use of AZA, MMF, tacrolimus, angiotensin converting enzyme inhibitors (ACEI), but not with cyclosporine (CSA) use. Post-transplant hemolytic uremic syndrome has been reported with orthoclone anti T-cell antibody (OKT3), CSA and tacrolimus therapy. Viral infections including cytomegalovirus, Epstein-Barr virus and human parvovirus B19 have been reported to cause generalized marrow suppression. Management of severe anemia associated with immunosuppressive drugs generally requires lowering the dose, drug substitution or, when possible, discontinuation of the drug. Because this topic has been incompletely studied, our recommendation as to the best immunosuppressive protocol after renal transplantation remains largely dependent on the clinical response of the individual patient. PMID- 12890003 TI - Risk of hearing impairment in pediatric liver transplant recipients: a single center study. AB - As survival rates following liver transplantation have increased, health care providers must assess the impact of transplantation on dimensions other than traditional medical measures. Hearing impairment may adversely impact social, emotional, cognitive, academic, and speech and language development. We hypothesized that children who undergo liver transplantation are at risk for hearing impairment due to exposure to ototoxic drugs. We conducted a review of 74 children who had undergone liver transplantation between December 1996 and September 2000 at Cincinnati Children's Hospital Medical Center. Hearing was assessed at discharge by an audiologist using age and developmentally appropriate techniques. The principal outcome measure was sensorineural hearing impairment. Independent variables were age at transplantation, United Network for Organ Sharing (UNOS) status at transplantation, primary diagnosis, post-transplant length of hospital stay, days of treatment with aminoglycosides, and days of treatment with loop diuretics. Eleven of 74 children (15%) had sensorineural hearing loss, of whom four had severe to profound hearing loss. Multivariate analyses showed that the adjusted relative risk for hearing loss in patients with hepatoblastoma was 66 and that there was a 5% increase risk for hearing loss for each additional day of hospitalization. Age at transplantation, UNOS status, and days of treatment with loop diuretics or aminoglycosides did not achieve significance in the model. Sensorineural hearing impairment occurs in a subset of pediatric patients following liver transplantation. Patients with hepatoblastoma or those who experience prolonged hospitalization after transplantation are at increased risk. Our observations are of particular importance for pediatric liver transplant recipients since the median age at transplantation is 12-18 months, a critical period for language acquisition. PMID- 12890004 TI - Renal function following liver transplantation for unresectable hepatoblastoma. AB - Combination of cyclosporine (CsA) and tacrolimus immunosuppression post-liver transplantation (LT) and the chemotherapeutic drugs used to treat hepatoblastoma (HB), are nephrotoxic. We aimed to determine the severity and duration of nephrotoxicity in children following LT for unresectable HB. We reviewed all children undergoing LT for unresectable HB at the Liver Unit, Birmingham Children's Hospital, UK, from 1991 to July 2000. Thirty-six children undergoing LT for biliary atresia, matched for age and sex, were selected as controls to compare pre- and post-LT renal function. Renal function was determined by estimation of glomerular filtration rate (eGFR) derived from plasma creatinine using Schwartz's formula. Twelve children with HB (mean age of diagnosis 33 months) who underwent LT (mean age 47 months) and 36 controls (mean age of LT 34 months) were studied. CsA was the main immunosuppressive drug used in each group. The median eGFR before, and at 3, 6, 12, 24 and 36 months after LT in HB group was significantly lower than controls (93 vs. 152, 66 vs. 79, 62 vs. 86, 66 vs. 87, 64 vs. 94, 53 vs. 90 mL/min/1.73 m2, respectively; 0.01 < p < 0.03). The reductions in the median eGFR of both the HB group and controls before and at 36 months after LT were 49 and 41%, respectively. At 36 months after LT, there was a trend for partial recovery of the eGFR in the controls but not in the HB group. Children who underwent LT for unresectable HB had renal dysfunction before transplantation that persisted for 36 months after LT. PMID- 12890005 TI - Lymphocyte data in Epstein-Barr-virus induced post-transplant lymphoproliferative disorder treated by rituximab. AB - Viral infection is an important cause of morbidity and mortality in the post allograft period. Recently, a new therapeutic approach was developed in post transplant lymphoproliferative disorder (PTLD) induced by Epstein-Barr virus (EBV): the anti-CD20 monoclonal antibody or rituximab. We performed a single center study on the treatment effectiveness of rituximab in three EBV-induced PTLD and evaluated biologic data, such as T and B lymphocytes count, during PTLD development and treatment. Before PTLD treatment, blood cell profile showed a severe T lymphopenia with a progressive increase of CD8+ cells and B lymphopenia. Secondly, during treatment, there appeared a T response, as in primary EBV, and a regressive B lymphopenia. PMID- 12890006 TI - Longitudinal evaluation of the pharmacokinetics of cyclosporin microemulsion (Neoral) in pediatric renal transplant recipients and assessment of C2 level as a marker for absorption. AB - BACKGROUND: There are important differences in CsA pharmacokinetics between adult and pediatric patients, such that pharmacokinetic data can not necessarily be extrapolated from the adult to the pediatric setting. Research in adult renal transplant patients has shown that adequate cyclosporin exposure (AUC0-4) in the first week post-transplant is important for successful clinical outcome, and that cyclosporin concentration at 2 h post-dose (C2) provides the optimal single-time point marker for AUC0-4. Clinically, dose management based on C2 level results in a low incidence of acute rejection in the adult renal transplant population. The study reported here undertook pharmacokinetic profiling in de novo renal transplant patients over a period of 6 months and retrospectively assessed alternative monitoring strategies based on pharmacokinetic findings and clinical outcomes. METHODS: This open-label, observational, prospective study was carried out at four UK transplant centers over a period of 6 months in pediatric de novo renal transplant recipients receiving the microemulsion formulation of cyclosporin (Neoral) according to local protocol. Twelve-hour pharmacokinetic profiles (8-16 blood samples each) were performed on days 5 and 14 and at weeks 4, 13 and 26 post-transplant. RESULTS: Thirty-two patients were recruited (median age 10 yr, range 3-18 yr). At 6 months, patient survival was 100% and graft survival was 91%. The incidence of clinically determined acute rejection was 41% (13 of 32). Six patients discontinued Neoral before 6 months: three due to graft loss, one due to rejection, one due to renal toxicity and one due to hypertrichosis. At all time points studied, C2 correlated more closely with AUC0 4 and with AUC0-12 than did the pre-dose cyclosporin concentration (C0, or trough). Patients achieving C2 > 1.5 microg/mL by the fifth postoperative day experienced no acute rejection in the first 6 months, compared with a 50% rejection rate among patients with C2 < 1.5 microg/mL (P < 0.05). Binary logistic regression analysis showed that C2 level >1.7 microg/mL was associated with approximately 90% probability of freedom from acute rejection. Analysis of renal function across patients grouped according to cyclosporine exposure (AUC0-4, C2) showed no adverse effects of higher/increased exposure on creatinine or GFR. CONCLUSIONS: C2 level provides a more reliable marker for CsA exposure than C0 in pediatric renal transplant recipients, and is more closely predictive of acute rejection risk. A C2 target of 1.7 microg/mL appears appropriate in this population during the immediate post-transplant period in order to maximize clinical benefit. PMID- 12890007 TI - Timed Pediatric Risk of Mortality Scores predict outcomes in pediatric liver transplant recipients. AB - More reliable methods are needed to identify children at risk for poor outcomes following liver transplantation. The Pediatric Risk of Mortality (PRISM) Score is a physiology-based scoring system used to quantify risk of mortality in pediatric intensive care unit (ICU) populations. We evaluated the PRISM Score as a predictor of outcomes including survival in the pediatric liver transplant (LT) population. We retrospectively reviewed the records of 67 consecutive LTs performed between August 1997 and February 2000 at an urban, tertiary children's hospital in Chicago, IL, USA. Four PRISM Scores were calculated to determine which periods were most meaningful. A Classic PRISM Score was calculated during first 24 h of ICU admission, and three PRISM Scores were timed with the patient's transplant: a pre-LT PRISM Score (24 h prior to transplant whether in ICU or not), a 24-h post-LT PRISM Score and a 48-h post-LT PRISM Score. These PRISM Scores and other predictors including transplant number, UNOS status and PELD Score were compared with outcomes including survival using univariate methods. The pre-LT, the 24- and the 48-h PRISM Score were associated with the post-LT number of ventilated days (p < 0.05), ICU days (p < 0.05) and with 1-yr survival (p < 0.04). The PRISM Scores were not related to the post-LT hospital length of stay (LOS) or to 1-yr re-transplantation. The PELD Score correlated with the post LT hospital LOS, but was not associated with mortality or with the ICU LOS. A patient's UNOS status and Classic PRISM Score were not associated with any of the outcomes measured. PRISM Scores are valid predictors of outcome including survival in pediatric LT recipients. These findings help to demonstrate the importance in this population of a patient's general physiologic condition and its influence on the overall hospital course and survival. PMID- 12890008 TI - Quantitative analysis of CMV DNA in children the first year after liver transplantation. AB - CMV infection is a major problem after solid organ transplantation especially in children where primary infection is more common than in adults. Early diagnosis is critical and might be facilitated by quantitative analysis of CMV DNA in blood. In this retrospective study of 18 children who had a liver transplantation 1995-2000, serum samples were analysed by Cobas Amplicor Monitor (Roche). Four patients developed symptomatic CMV infection at a mean time of 4 wk after transplantation. They showed maximum CMV DNA levels in serum of 26 400, 1900, 1300 and 970 copies/mL, respectively. In comparison, CA Monitor was positive, at a low level (415 copies/mL), in one of 11 patients with asymptomatic (4) or latent (7) infection. CMV IgM was detected at significant levels (> or =1/80) in all four patients with symptomatic, and in one with asymptomatic CMV infection. Eight patients were given one or several courses of ganciclovir. Five of these lacked symptoms of CMV disease, and had low (415 copies/mL) or undetectable CMV DNA in serum. The data suggest that quantitative analysis of CMV DNA may be of value in early identification of CMV disease and for avoiding unnecessary antiviral treatment. PMID- 12890009 TI - Alterations of cyclosporin A metabolism induced by mycophenolate mofetil. AB - Unexpectedly lower CsA 2-h levels were found in patients who received additional immunosuppression by MMF in our center. The aim of this study is to prove whether there are differences in CsA metabolization when MMF is added to treatment. In 33 children who received an immunosuppression of prednisolone and CsA as well as in 15 children who were treated with additional MMF, C2 and C0 were taken. In 11 children, full AUC of CsA were obtained. C2 levels differed significantly (p < 0.05) between patients treated with (617 +/- 230 ng/mL) and without MMF (750 +/- 271 ng/mL) but with similar CsA dosages. C2 correlated better with the AUC than CsA levels at other time points with r = 0.95. The equation AUC = 139 + 6.17 x C2 was calculated to match best in a C2-limited sampling strategy. This formula proved to be safer than equations that had been published before. According to our findings, we suspect that MMF alters the CsA metabolism. When MMF is used in pediatric transplant recipients, either target values of CsA have to be changed or patients may require higher doses of CsA. PMID- 12890010 TI - Monitoring and modulation of Epstein-Barr virus loads in pediatric transplant patients. AB - A major risk faced by bone-marrow and solid organ transplant patients is the development of post-transplant lymphoproliferative disease or post-transplant lymphoma (PTLD). In pediatric transplantation, PTLD onset is often associated with a rapid rise in Epstein-Barr virus (EBV) load in peripheral blood mononuclear cells (PBMC). We have analyzed EBV viral loads in PBMC over time using real-time quantitative PCR in 56 patients, 19 of which have been followed for more than 1 year. In nine patients; eight bone marrow (BMT) and one kidney transplant, PTLD was associated with a rapid rise in viral load, exceeding 1 x 10(5) EBV genomes/microg of PBMC-derived DNA. Four of these patients exceeded 1 x 10(6) EBV genomes/microg PBMC DNA. All patients with viral loads exceeding 1 x 10(5) EBV genomes/microg PBMC DNA were clearly at high risk for transplant associated mortality, with only six of nine surviving. Importantly, only one of these deaths was directly attributable to EBV. A second elevated state of EBV load, defined as exceeding 2 x 10(4) EBV genomes/microg PBMC, was seen in a total of 12 BMT, kidney, heart, and liver transplant patients. These patients did not appear to be at immediate lethal risk for PTLD and one EBV-attributable death was found in this group as well. Thirty-four transplant patients whose EBV viral load oscillated from undetectable to 10 000 EBV genomes/microg PBMC DNA are reported as well. The threshold for normal EBV viral load based on our combined experience with viral load analysis is defined as 1 x 10(4) EBV genomes/microg PBMC DNA. The ability to rapidly analyze EBV load allows rapid changes in viral load, such as those that occur with PTLD onset, and the impact of anti-CD20 antibody therapy to be rapidly detected. PMID- 12890011 TI - Non-cardiogenic pulmonary edema during basiliximab induction in three adolescent renal transplant patients. AB - BACKGROUND: Introduction of the anti-CD-25 mAb basiliximab into renal transplant protocols has reduced the incidence of acute rejection. However, its side-effect profile is still unfolding. We report three adolescents who developed severe non cardiogenic PE within 2 days of renal transplantation. METHODS: Pretransplant cardiorespiratory evaluation was normal in all cases. Transplant immunosuppression consisted of basiliximab induction, corticosteroids, and tacrolimus. Patients received standard fluid management during and after the transplant surgery. CASE REPORTS: Patients 1 and 2 were 17- and 21-yr-old females. Pretransplant Hct values were 35 and 25% respectively. Each received 5-L normal saline during surgery. EBL was 200 and 500 mL in patients 1 and 2, respectively. There was immediate post-operative diuresis. Both developed non cardiogenic PE by POD no. 2. BIPAP and PRVC were administered respectively. In both cases PE resolved within 1 wk. Patient 3 was a 19-yr-old male with pretransplant Hct of 43% who received a cadaveric renal transplant after 23.5-h cold-ischemia; 3.5 L normal saline was given during surgery. EBL was 100 mL. Non cardiogenic PE ensued on POD no. 2 warranting assisted ventilation. The patient died following a sudden cardiopulmonary arrest on POD no. 3. CONCLUSIONS: Potential mechanisms for the development of PE include cytokine release from basiliximab with increased capillary permeability, volume overload and ischemic reperfusion injury. Improved awareness of this potential complication, prudent fluid management, and efforts to minimize graft-ischemia are recommended to prevent further cases. PMID- 12890012 TI - Changing trends in pediatric transplantation: 2001 Annual Report of the North American Pediatric Renal Transplant Cooperative Study. AB - The North American Pediatric Renal Transplant Cooperative Study has collected clinical information on children undergoing a renal transplantation since 1987. This cooperative group now includes over 150 participating medical centers in the United States, Canada, Mexico, and Costa Rica. This report covers the years from 1987 through 2001 and includes data on 7545 renal transplants in 6878 patients. This report demonstrates changing trends in many areas of pediatric transplantation including increasing numbers of African American and Hispanic children receiving transplantation, remarkable improvements in the rate of acute rejection, rejection reversal, and short- and long-term allograft survival. In the most recent cohorts of patients, we now see that 1-yr allograft survival is no different in cadaver donor compared to living donor recipients and in infants compared to all other age groups. However, this analysis also reveals areas of continued challenges including inferior outcomes in African American and adolescent populations, chronic rejection, and the adverse effects of immunosuppression. PMID- 12890013 TI - Regulation of fungal gene expression via short open reading frames in the mRNA 5'untranslated region. AB - We review how the expression of fungal mRNAs can be controlled by ribosome interactions with short upstream open reading frames (uORFs) within the 5'untranslated region. The efficiency of uAUG recognition modulates the impact of a uORF but steps during and after translation of the uORF also influence uORF function. The post-termination behaviour of ribosomes, therefore, plays a major role in determining the expression level of these main ORFs. Translation of a uORF can produce a cis-acting peptide that causes effector molecule-dependent stalling of the ribosomes at the end of the uORF. In other cases it is the length or position, or other features of the uORF, rather than the peptide it encodes, that determine the efficiency with which ribosomes reinitiate translation downstream of it. Whether the form of the ribosome that resumes scanning after termination is the 40S subunit alone or the entire 80S ribosome is not known. Translation of the uORF can also control gene expression by affecting the stability of the mRNA. Finally, trans-acting factors may participate in the regulatory mechanisms. Future work will need not only to provide more information on the mechanisms underlying the known cases of uORF-mediated control but also to define the full complement of uORF-containing mRNAs in at least one fungal organism. PMID- 12890014 TI - Touch and go: tying TonB to transport. AB - The TonB system of Gram-negative bacteria appears to exist for the purpose of transducing the protonmotive force energy from the cytoplasmic membrane, where it is generated, to the outer membrane, where it is needed for active transport of iron siderophores, vitamin B12 and, in pathogens, iron from host-binding proteins. In this review, we bring the reader up to date on the developments in the field since the authors each wrote reviews in this journal in 1990. PMID- 12890015 TI - TgSUB2 is a Toxoplasma gondii rhoptry organelle processing proteinase. AB - All parasites in the phylum Apicomplexa, including Toxoplasma gondii and Plasmodium falciparum, contain rhoptries, specialized secretory organelles whose contents are thought to be essential for successful invasion of host cells. Serine proteinase inhibitors have been reported to block host cell invasion by both T. gondii and P. falciparum. We describe the cloning and characterization of TgSUB2, a subtilisin-like serine proteinase, from T. gondii. Like its closest homologue P. falciparum PfSUB-2, TgSUB2 is predicted to be a type I transmembrane protein. Disruption of TgSUB2 was unsuccessful implying that TgSUB2 is an essential gene. TgSUB2 undergoes autocatalytic processing as it traffics through the secretory pathway. TgSUB2 localizes to rhoptries and associates with rhoptry protein ROP1, a potential substrate. A sequence within TgSUB2 with homology to the ROP1 cleavage site (after Glu) was identified and mutated by site-directed mutagenesis. This mutation abolished TgSUB2 autoprocessing suggesting that TgSUB2 is a rhoptry protein maturase with similar specificity to the ROP1 maturase. Processing of secretory organelle contents appears to be ubiquitous among the Apicomplexa. As subtilases are present in genomes of all the Apicomplexa sequenced to date, subtilases may represent a novel chemotherapeutic target. PMID- 12890016 TI - A strand-specific model for chromosome segregation in bacteria. AB - Chromosome separation and segregation must be executed within a bacterial cell in which the membrane and cytoplasm are highly structured. Here, we develop a strand specific model based on each of the future daughter chromosomes being associated with a different set of structures or hyperstructures in an asymmetric cell. The essence of the segregation mechanism is that the genes on the same strand in the parental cell that are expressed together in a hyperstructure continue to be expressed together and segregate together in the daughter cell. The model therefore requires an asymmetric distribution of classes of genes and of binding sites and other structures on the strands of the parental chromosome. We show that the model is consistent with the asymmetric distribution of highly expressed genes and of stress response genes in Escherichia coli and Bacillus subtilis. The model offers a framework for interpreting data from genomics. PMID- 12890017 TI - Transition from reversible to irreversible attachment during biofilm formation by Pseudomonas fluorescens WCS365 requires an ABC transporter and a large secreted protein. AB - We report the identification of an ATP-binding cassette (ABC) transporter and an associated large cell-surface protein that are required for biofilm formation by Pseudomonas fluorescens WCS365. The genes coding for these proteins are designated lap for large adhesion protein. The LapA protein, with a predicted molecular weight of approximately 900 kDa, is found to be loosely associated with the cell surface and present in the culture supernatant. The LapB, LapC and LapE proteins are predicted to be the cytoplasmic membrane-localized ATPase, membrane fusion protein and outer membrane protein component, respectively, of an ABC transporter. Consistent with this prediction, LapE, like other members of this family, is localized to the outer membrane. We propose that the lapEBC-encoded ABC transporter participates in the secretion of LapA, as strains with mutations in the lapEBC genes do not have detectable LapA associated with the cell surface or in the supernatant. The lap genes are conserved among environmental pseudomonads such as P. putida KT2440, P. fluorescens PfO1 and P. fluorescens WCS365, but are absent from pathogenic pseudomonads such as P. aeruginosa and P. syringae. The wild-type strain of P. fluorescens WCS365 and its lap mutant derivatives were assessed for their biofilm forming ability in static and flow systems. The lap mutant strains are impaired in an early step in biofilm formation and are unable to develop the mature biofilm structure seen for the wild-type bacterium. Time-lapse microscopy studies determined that the lap mutants are unable to progress from reversible (or transient) attachment to the irreversible attachment stage of biofilm development. The lap mutants were also found to be defective in attachment to quartz sand, an abiotic surface these organisms likely encounter in the environment. PMID- 12890018 TI - Biphasic intracellular expression of Staphylococcus aureus virulence factors and evidence for Agr-mediated diffusion sensing. AB - Staphylococcus aureus invades a variety of mammalian cells and escapes from the endosome to multiply in the cytoplasm. We had previously hypothesized that the molecular events leading to escape of S. aureus from the endosome involved the Agr virulence factor regulatory system. In this report we demonstrate that temporal changes in intracellular activation of the Agr regulon correlates with expression of membrane active toxins. Also, the initial expression of Agr by even small numbers of staphylococci resulted in the permeabilization of the endosomal membrane and the eventual escape of bacteria into the cytoplasm by 3 h post invasion. After Agr downregulation, a second peak of expression coincided with increased permeability of the host cell membrane. In contrast to the parental strain, an Agr-mutant was unable to escape into the cytoplasm and was observed in intact endosomes as late as 5 h post invasion. These data provide evidence that staphylococcal virulence factor production during invasion of host cells is mediated by an Agr-dependent process that is most accurately described in the context of diffusion sensing. PMID- 12890020 TI - Characterization of the roles of NikR, a nickel-responsive pleiotropic autoregulator of Helicobacter pylori. AB - The Helicobacter pylori genome contains a gene (hp1338 or nikR) that encodes a nickel-dependent regulator that is homologous to the Escherichia coli nickel responsive regulator, NikR. The H. pylori nikR product acts as a pleiotropic metal-dependent regulator. We constructed a non-polar isogenic mutant deleted for the nikR gene. NikR was essential for the survival of H. pylori in the presence of high nickel and cobalt ion concentrations in vitro. We screened a DNA macroarray for genes that were differentially expressed in parental and nikR deficient H. pylori strains grown in the presence of excess nickel. We found that H. pylori NikR mediates the expression of nickel-activated and -repressed genes. In the presence of excess nickel, NikR activated the transcription of ureA-ureB (hp72-73), nixA (hp1077 ), copA2 (hp1072), hpn (hp1427 ) and hpn-like (hp1432) genes and repressed the expression of genes encoding proteins involved in ferric iron uptake and storage [pfr (hp0653), fur (hp1027 ), frpB4 (hp1512), exbB/exbD (hp1339-1340), ceuE (hp1561)], motility [cheV (hp616), flaA (hp0601), flaB (hp0115 )], stress responses [hrcA-grpE-dnaK (hp111-110-109)] and encoding outer membrane proteins [omp11(hp0472), omp31 (hp1469), omp32 (hp1501)]. Slot blot DNA/RNA hybridization experiments using RNA from three independent bacterial cultures confirmed the transcriptome data for 10 selected genes. The results of gel shift experiments using purified native NikR, beta-galactosidase assays with the region between nikR and the exbB/exbD divergent operon, and the study of exbB gene expression using a gentamicin/apramycin reporter gene in H. pylori indicated that NikR is an autorepressor that binds to this intergenic region and also controls the expression of the exbB/exbD/tonB operon, which provides energy for ferric iron uptake. Thus, as previously suggested for Fur in H. pylori, NikR appears to be a global regulator of the metabolism of some divalent cations within a highly complex regulated network. PMID- 12890021 TI - Coxiella burnetii express type IV secretion system proteins that function similarly to components of the Legionella pneumophila Dot/Icm system. AB - Coxiella burnetii is an obligate intracellular pathogen that replicates in large endocytic vacuoles. Genomic sequence data indicate that 21 genes encoding products that are similar to components of the Legionella pneumophila Dot/Icm type IV secretion system are located on a contiguous 35 kb region of the Coxiella chromosome. It was found that several dot/icm genes were expressed by Coxiella during host cell infection and that dot/icm gene expression preceded the formation of large replicative vacuoles. To determine whether these genes encode a functional type IV secretion system, we have amplified the Coxiella dotB, icmQ, icmS and icmW genes and produced the encoded proteins in Legionella mutants in which the native copy of each gene had been deleted. The Coxiella dotB, icmS and icmW products restored dot/icm-dependent growth of Legionella mutants in eukaryotic host cells. The Coxiella IcmQ protein and the Legionella IcmR protein did not interact, which could explain why the Coxiella icmQ gene was unable to restore growth to a Legionella icmQ mutant. Thus, Coxiella encodes functional components of a type IV secretion system expressed in vivo that is mechanistically related to the Legionella Dot/Icm apparatus. These studies suggest that a dot/icm-related secretion system could play an important role in creating the specialized vacuole that supports Coxiella replication. PMID- 12890019 TI - Pathogenic Leptospira species express surface-exposed proteins belonging to the bacterial immunoglobulin superfamily. AB - Proteins with bacterial immunoglobulin-like (Big) domains, such as the Yersinia pseudotuberculosis invasin and Escherichia coli intimin, are surface-expressed proteins that mediate host mammalian cell invasion or attachment. Here, we report the identification and characterization of a new family of Big domain proteins, referred to as Lig (leptospiral Ig-like) proteins, in pathogenic Leptospira. Screening of L. interrogans and L. kirschneri expression libraries with sera from leptospirosis patients identified 13 lambda phage clones that encode tandem repeats of the 90 amino acid Big domain. Two lig genes, designated ligA and ligB, and one pseudogene, ligC, were identified. The ligA and ligB genes encode amino terminal lipoprotein signal peptides followed by 10 or 11 Big domain repeats and, in the case of ligB, a unique carboxy-terminal non-repeat domain. The organization of ligC is similar to that of ligB but contains mutations that disrupt the reading frame. The lig sequences are present in pathogenic but not saprophytic Leptospira species. LigA and LigB are expressed by a variety of virulent leptospiral strains. Loss of Lig protein and RNA transcript expression is correlated with the observed loss of virulence during culture attenuation of pathogenic strains. High-pressure freeze substitution followed by immunocytochemical electron microscopy confirmed that the Lig proteins were localized to the bacterial surface. Immunoblot studies with patient sera found that the Lig proteins are a major antigen recognized during the acute host infection. These observations demonstrate that the Lig proteins are a newly identified surface protein of pathogenic Leptospira, which by analogy to other bacterial immunoglobulin superfamily virulence factors, may play a role in host cell attachment and invasion during leptospiral pathogenesis. PMID- 12890022 TI - Multiple transporters associated with malaria parasite responses to chloroquine and quinine. AB - Mutations and/or overexpression of various transporters are known to confer drug resistance in a variety of organisms. In the malaria parasite Plasmodium falciparum, a homologue of P-glycoprotein, PfMDR1, has been implicated in responses to chloroquine (CQ), quinine (QN) and other drugs, and a putative transporter, PfCRT, was recently demonstrated to be the key molecule in CQ resistance. However, other unknown molecules are probably involved, as different parasite clones carrying the same pfcrt and pfmdr1 alleles show a wide range of quantitative responses to CQ and QN. Such molecules may contribute to increasing incidences of QN treatment failure, the molecular basis of which is not understood. To identify additional genes involved in parasite CQ and QN responses, we assayed the in vitro susceptibilities of 97 culture-adapted cloned isolates to CQ and QN and searched for single nucleotide polymorphisms (SNPs) in DNA encoding 49 putative transporters (total 113 kb) and in 39 housekeeping genes that acted as negative controls. SNPs in 11 of the putative transporter genes, including pfcrt and pfmdr1, showed significant associations with decreased sensitivity to CQ and/or QN in P. falciparum. Significant linkage disequilibria within and between these genes were also detected, suggesting interactions among the transporter genes. This study provides specific leads for better understanding of complex drug resistances in malaria parasites. PMID- 12890023 TI - Host plant-dependent phenotypic reversion of Ralstonia solanacearum from non pathogenic to pathogenic forms via alterations in the phcA gene. AB - Ralstonia solanacearum is a plant pathogenic bacterium that undergoes a spontaneous phenotypic conversion (PC) from a wild-type pathogenic to a non pathogenic form. PC is often associated with mutations in phcA, which is a key virulence regulatory gene. Until now, reversion to the wild-type pathogenic form has not been observed for PC variants and the biological significance of PC has been questioned. In this study, we characterized various alterations in phcA (eight IS element insertions, three tandem duplications, seven deletions and a base substitution) in 19 PC mutants from the model strain GMI1000. In five of these variants, reversion to the pathogenic form was observed in planta, while no reversion was ever noticed in vitro whatever culture media used. However, reversion was observed for a 64 bp tandem duplication in vitro in the presence of tomato root exudate. This is the first report showing a complete cycle of phenotypic conversion/reversion in a plant pathogenic bacterium. PMID- 12890024 TI - Prion protein gene polymorphisms in Saccharomyces cerevisiae. AB - The yeast Saccharomyces cerevisiae genome encodes several proteins that, in laboratory strains, can take up a stable, transmissible prion form. In each case, this requires the Asn/Gln-rich prion-forming domain (PrD) of the protein to be intact. In order to further understand the evolutionary significance of this unusual property, we have examined four different prion genes and their corresponding PrDs, from a number of naturally occurring strains of S. cerevisiae. In 4 of the 16 strains studied we identified a new allele of the SUP35 gene (SUP35delta19) that contains a 19-amino-acid deletion within the N terminal PrD, a deletion that eliminates the prion property of Sup35p. In these strains a second prion gene, RNQ1, was found to be highly polymorphic, with eight different RNQ1 alleles detected in the six diploid strains studied. In contrast, for one other prion gene (URE2) and the sequence of the NEW1 gene encoding a PrD, no significant degree of DNA polymorphism was detected. Analysis of the naturally occurring alleles of RNQ1 and SUP35 indicated that the various polymorphisms identified were associated with DNA tandem repeats (6, 12, 33, 42 or 57 bp) within the coding sequences. The expansion and contraction of DNA repeats within the RNQ1 gene may provide an evolutionary mechanism that can ensure rapid change between the [PRION+] and [prion-] states. PMID- 12890025 TI - Expression and regulation of the crucial plant-like ferredoxin of cyanobacteria. AB - The Synechocystis fedI gene (petF, ssl0020) was found to be strongly expressed under the negative control of H2O2 or heavy metals, and the positive control of light fluence (regulation dependent on active photosynthesis) or carbon availability [under the control of NdhR, the regulator of the ndh3 operon encoding NAD(P)H dehydrogenase subunits]. The basic and constitutive promoter (BP) of fedI extending from -62 to +25 (relative to the transcription start point) is weakly active, presumably because it harbours a long (30 bp) spacer between the two crucial motifs: the -10 box (5'-TAgtAT-3', -13 to -8) and the ' 35' box (5'-TTGctA-3', -49 to -44). BP strength is strongly enhanced by the two upstream regions, -113 to -82 and -151 to -114, mediating the 30-fold constitutive stimulation and the fourfold light activation respectively. Three well-conserved transcriptional elements were characterized for the first time, namely the -19 box (5'-TTTT-3') that is essential to transcription, and the two twice repeated elements that are both critical to light induction: the TTGyCA-3' box (-35 to -30, and -125 to -120) and the 5'-ATTTyA-3' box (-55 to -50, and -134 to -129). That two of these light induction motifs (5'-TTGtCA-3', -35 to -30; 5' ATTTcA-3', -55 to -50) occur in the constitutive BP promoter indicate that in the fedI gene light activation and transcription per se are closely interacting. Interestingly, the fedI gene from marine strains was found to lack the three transcriptional elements presently described, as well as the 5'-AGGA-3' Shine Dalgarno sequence, which are all conserved among the fedI from non-marine strains. PMID- 12890026 TI - Microcin E492 antibacterial activity: evidence for a TonB-dependent inner membrane permeabilization on Escherichia coli. AB - The mechanism of action of microcin E492 (MccE492) was investigated for the first time in live bacteria. MccE492 was expressed and purified to homogeneity through an optimized large-scale procedure. Highly purified MccE492 showed potent antibacterial activity at minimal inhibitory concentrations in the range of 0.02 1.2 microM. The microcin bactericidal spectrum of activity was found to be restricted to Enterobacteriaceae and specifically directed against Escherichia and Salmonella species. Isogenic bacteria that possessed mutations in membrane proteins, particularly of the TonB-ExbB-ExbD complex, were assayed. The microcin bactericidal activity was shown to be TonB- and energy-dependent, supporting the hypothesis that the mechanism of action is receptor mediated. In addition, MccE492 depolarized and permeabilized the E. coli cytoplasmic membrane. The membrane depolarization was TonB dependent. From this study, we propose that MccE492 is recognized by iron-siderophore receptors, including FepA, which promote its import across the outer membrane via a TonB- and energy-dependent pathway. MccE492 then inserts into the inner membrane, whereupon the potential becomes destabilized by pore formation. Because cytoplasmic membrane permeabilization of MccE492 occurs beneath the threshold of the bactericidal concentration and does not result in cell lysis, the cytoplasmic membrane is not hypothesized to be the sole target of MccE492. PMID- 12890027 TI - The pair of arginine codons AGA AGG close to the initiation codon of the lambda int gene inhibits cell growth and protein synthesis by accumulating peptidyl tRNAArg4. AB - To analyse the mechanism by which rare codons near the initiation codon inhibit cell growth and protein synthesis, we used the bacteriophage lambda int gene or early codon substitution derivatives. The lambda int gene has a high frequency of rare ATA, AGA and AGG codons; two of them (AGA AGG) located at positions 3 and 4 of the int open reading frame (ORF). Escherichia coli pth (rap) cells, which are defective in peptidyl-tRNA hydrolase (Pth) activity, are more susceptible to the inhibitory effects of int expression as compared with wild-type cells. Cell growth and Int protein synthesis were enhanced by overexpression of Pth and tRNAArg4 cognate to AGG and AGA but not of tRNAIle2a specific for ATA. The increase of Int protein synthesis also takes place when the rare arginine codons AGA and AGG at positions 3 and 4 are changed to common arginine CGT or lysine AAA codons but not to rare isoleucine ATA codons. In addition, overexpression of int in Pth defective cells provokes accumulation of peptidyl-tRNAArg4 in the soluble fraction. Therefore, cell growth and Int synthesis inhibition may be due to ribosome stalling and premature release of peptidyl-tRNAArg4 from the ribosome at the rare arginine codons of the first tandem, which leads to cell starvation for the specific tRNA. PMID- 12890028 TI - Staphostatins: an expanding new group of proteinase inhibitors with a unique specificity for the regulation of staphopains, Staphylococcus spp. cysteine proteinases. AB - A novel type of cysteine proteinase inhibitor (SspC) has been recently recognized in Staphylococcus aureus (Massimi, I., Park, E., Rice, K., Muller-Esterl, W., Sauder, D.N., and McGavin, M.J. (2002) J Biol Chem 277: 41770-41777). In this paper we have identified homologous proteins encoded in the genome of S. aureus and other coagulase-negative Staphylococci. Collectively we refer to these proteins as staphostatins as they specifically inhibit cysteine proteinases (staphopains) from Staphylococcus spp. The primary structure of staphostatins seems to be unique, although they resemble cystatins in size (105-108 residues). Recombinant staphostatin A, a product of the scpB gene and staphostatin B (SspC) from S. aureus have been characterized in details. Similar to the cystatins, the staphostatins interact specifically with their target proteinases forming tight and stable non-covalent complexes, staphostatin A with staphopain A and staphostatin B with staphopain B. However, in contrast to the cystatins, each of which inhibits broad range of cathepsins, complex formation between staphostatin and staphopain appears to be exclusive, with no cross interaction observed. In addition, the activities of several tested cysteine proteinases of prokaryotic- and eukaryotic-origin were not affected by staphostatins. Such narrow specificity limited to staphopains is presumed to be required to protect staphylococcal cytoplasmic proteins from being degraded by prematurely activated/folded prostaphopains. This function is guaranteed through the unique co-expression of the secreted proteinase and the intracellular inhibitor from the same operon, and represents a unique mechanism of regulation of proteolytic activity in Gram positive bacteria. PMID- 12890029 TI - Phosphatidylethanolamine and phosphatidylglycerol are segregated into different domains in bacterial membrane. A study with pyrene-labelled phospholipids. AB - To detect and characterize membrane domains that have been proposed to exist in bacteria, two kinds of pyrene-labelled phospholipids, 2-pyrene-decanoyl phosphatidylethanolamine (PY-PE) and 2-pyrene-decanoyl-phosphatidylglycerol (PY PG) were inserted into Escherichia coli or Bacillus subtilis membrane. The excimerization rate coefficient, calculated from the excimer-to-monomer ratio dependencies on the probe concentration, was two times higher for PY-PE than for PY-PG at 37 degrees C. This was ascribed to different local concentrations rather than to differences in mobility. The extent of mixing between the two fluorescent phospholipids, estimated by formation of their heteroexcimer, was found very low both in E. coli and B. subtilis, in contrast to model membranes. In addition, these two pyrene derivatives exhibited different temperature phase transitions and different detergent extractability, indicating that the surroundings of these phospholipids in bacterial membrane differ in organization and order. Inhibition of protein synthesis, leading to condensation of nucleoid and presumably to dissipation of membrane domains, indeed resulted in increased formation of heteroexcimers, broadening of phase transitions and equal detergent extractability of both probes. It is proposed that in bacterial membranes these phospholipids are segregated into distinct domains that differ in composition, proteo-lipid interaction and degree of order; the proteo-lipid domain being enriched by PE. PMID- 12890030 TI - The Aspergillus nidulans metR gene encodes a bZIP protein which activates transcription of sulphur metabolism genes. AB - The identification, isolation and characterization of a new Aspergillus nidulans positive-acting gene metR, which encodes a transcriptional activator of sulphur metabolism, is reported. metR mutants are tight auxotrophs requiring methionine or homocysteine for growth. Mutations in the metR gene are epistatic to mutations in the negative-acting sulphur regulatory scon genes. The metR coding sequence is interrupted by a single intron of 492 bp which is unusually long for fungi. Aspergillus nidulans METR is a member of bZIP family of DNA-binding proteins. The bZIP domains of METR and the Neurospora crassa CYS3 transcriptional activator of sulphur genes are highly similar. Although Neurospora cys-3 gene does not substitute for the metR function, a chimeric metR gene with a cys-3 bZIP domain is able to transform the DeltametR mutant to methionine prototrophy. This indicates that METR recognizes the same regulatory sequence as CYS3. The metR gene is not essential, as deletion mutants are viable and have similar phenotype as point mutants. In contrast to the Neurospora cys-3, transcription of the metR gene was found to be regulated neither by METR protein nor by sulphur source. Transcription of metR gene is derepressed in the sconB2 mutant. Transcription of genes encoding sulphate permease, homocysteine synthase, cysteine synthase, ATP sulphurylase, and sulphur controller--sconB is strongly regulated by the metR gene product and depends on the character of the metR mutation and sulphur supplementation. PMID- 12890031 TI - Co-operative interactions control conjugative transfer of broad host-range plasmid RK2: full effect of minor changes in TrbA operator depends on KorB. AB - A network of circuits, with KorB and TrbA as key regulators, controls genes for conjugative transfer of broad host range plasmid RK2. To assess the importance of the TrbA regulon, mutational analysis was applied to the TrbA operator at the trbB promoter and then to other TrbA-regulated promoters in the tra region. All identified TrbA operators are submaximal; in the case of trbBp, a G to A transition that made the operator core a perfect palindrome increased repression by about 50% compared to the wild type. When this change was introduced into the RK2 genome, decreases in transfer frequency of up to three orders of magnitude were observed, with bigger effects when Escherichia coli was the donor compared to Pseudomonas putida. Western blotting showed a significant decrease in Trb protein levels. These effects were much greater than the effect of the mutation on repression by TrbA alone. When KorB was introduced into the reporter system, the effects were closer to those observed in the whole RK2 context. These results indicate that co-operativity, previously observed between TrbA and KorB, allows big changes in transfer gene expression to result from small changes in individual regulator activities. PMID- 12890032 TI - Distant cis-active sequences and sialic acid control the expression of fimB in Escherichia coli K-12. AB - The phase variation of type 1 fimbriation in Escherichia coli is controlled by the inversion of a 314 bp element of DNA, determined by FimB (switching in both directions) or FimE (switching from the ON-to-OFF orientation predominantly), and influenced by auxiliary factors IHF, Lrp and H-NS. The fimB gene is separated from the divergently transcribed yjhATS operon by a large (1.4 kbp) intergenic region of unknown function. Here, we show that fimB expression is regulated by multiple cis-active sequences that lie far upstream (>600 bp) of the transcription start sites for the recombinase gene. Two regions characterized further (regions 1 and 2) show sequence identity, and each coincides with a methylation-protected Dam (5'-GATC) site. Regions 1 and 2 apparently control fimB expression by an antirepression mechanism that involves additional sequences proximal to yjhA. Region 1 encompasses a 27 bp DNA sequence conserved upstream of genes known (nanAT ) or suspected (yjhBC) to be involved in sialic acid metabolism, and we show that FimB expression and recombination are suppressed by N-acetylneuraminic acid. We propose that E. coli recognizes the amino sugars as a harbinger of potential host defence activation, and suppresses the expression of type 1 fimbriae in response. PMID- 12890033 TI - Three pathways for trehalose metabolism in Corynebacterium glutamicum ATCC13032 and their significance in response to osmotic stress. AB - Genome scanning of Corynebacterium glutamicum ATCC13032 revealed the presence of five different genes encoding enzymes belonging to three putative trehalose biosynthesis pathways (OtsAB, TreYZ, TreS). The function of the different pathways and of trehalose as an osmoprotectant was studied by characterizing several strains defective for individual trehalose biosynthetic routes. Trehalose synthesis was shown to increase upon hyperosmotic conditions. Cytoplasmic trehalose levels varied considerably depending on kind and accessibility of carbon and nitrogen sources. In contrast to other organisms, osmoregulated trehalose synthesis in C. glutamicum is mediated by the TreYZ and not by the OtsAB pathway. Irrespective of their significance for the osmotic response, otsA and treS were upregulated at the transcriptional level after hyperosmotic shock. In vivo, TreS-mediated trehalose synthesis only occurred if maltose was used as the carbon source. In vitro, TreS catalysed the conversion of maltose into trehalose and, conversely, trehalose into maltose. As the reaction seems to be near equilibrium, TreS appears to be important for trehalose degradation rather than synthesis because a 1000-fold excess of trehalose to maltose was detected in the cytoplasm. Also, evidence is given that both the OtsAB and the TreYZ pathways are involved, but not essential, in supplying trehalose for mycolic acid biosynthesis. PMID- 12890034 TI - The BceRS two-component regulatory system induces expression of the bacitracin transporter, BceAB, in Bacillus subtilis. AB - BceA and bceB encode a nucleotide-binding domain (NBD) and membrane-spanning domain (MSD) subunit, respectively, of an ATP-binding cassette (ABC) transporter in Bacillus subtilis. Disruption of these genes resulted in hypersensitivity to bacitracin, a peptide antibiotic that is non-ribosomally synthesized in some strains of Bacillus. Northern hybridization analyses showed that expression of the bceAB operon is induced by bacitracin present in the growth medium. The bceRS genes encoding a two-component regulatory system are located immediately upstream of bceAB. Deletion analyses of the bceAB promoter together with DNase I footprinting experiments revealed that a sensor kinase, BceS, responds to extracellular bacitracin either directly or indirectly and transmits a signal to a cognate response regulator, BceR. The regulator binds directly to the upstream region of the bceAB promoter and upregulates the expression of bceAB genes. The bcrC gene product is additionally involved in bacitracin resistance. The expression of bcrC is dependent on the ECF sigma factors, sigmaM and sigmaX, but not on the BceRS two-component system. In view of these results, possible roles of BceA, BceB and BcrC in bacitracin resistance of B. subtilis 168 are discussed. PMID- 12890035 TI - An intramolecular disulphide bond reduces the efficacy of a lipoprotein plasma membrane sorting signal. AB - To study lipoprotein sorting in Escherichia coli, we devised a novel screen in which sensitivity or resistance to bacteriophage T5 and colicin M reflects the membrane localization of the bacteriophage T5-encoded lipoprotein Llp, which inactivates the outer membrane (OM) T5 receptor (FhuA). When processed by lipoprotein signal peptidase, Llp has a serine at position +2, immediately after the fatty acylated N-terminal cysteine. As predicted by the '+2 lipoprotein sorting rule' that determines the localization of lipoproteins in the cell envelope, Llp is located in the OM. However, contrary to expectations, when serine +2 was replaced by aspartate, the canonical plasma membrane lipoprotein retention signal, Llp was still > or =40% targeted to the OM and protected cells against colicin M and phage T5. OM association of this Llp derivative was abolished when a peptide spacer was inserted between the aspartate and the rest of Llp or when the formation of an intramolecular disulphide bond in Llp was prevented by substituting one or other of the cysteines involved. Furthermore, analysis of a MalE-Llp hybrid protein with or without a lipid moiety demonstrated that fatty acylation of Llp is essential for its OM association and for protection against colicin M and bacteriophage T5. These data suggest (i) that phage-encoded Llp uses the endogenous E. coli Lol pathway for lipoprotein sorting to the OM and (ii) that the conformation of a lipoprotein can affect its sorting within the cell envelope. PMID- 12890036 TI - Circadian profile of cardiac autonomic nervous modulation in healthy subjects: differing effects of aging and gender on heart rate variability. AB - Circadian Profile of Heart Rate Variability. INTRODUCTION: Although heart rate variability (HRV) has been established as a tool to study cardiac autonomic activity, almost no data are available on the circadian patterns of HRV in healthy subjects aged 20 to 70 years. METHODS AND RESULTS: We investigated 166 healthy volunteers (81 women and 85 men; age 42 +/- 15 years, range 20-70) without evidence of cardiac disease. Time-domain HRV parameters were determined from 24-hour Holter monitoring and calculated as hourly mean values and mean 24 hour values. All volunteers were fully mobile, awoke around 7 A.M., and had 6 to 8 hours of sleep. Circadian profiles of vagus-associated HRV parameters revealed a marked day-night pattern, with a peak at nighttime and a plateau at daytime. The characteristic nocturnal peak and the day-night amplitude diminished with aging by decade. Estimates of overall HRV (geometric triangular index [TI], SD of NN intervals [SDNN]) and long-term components of HRV (SD of the averages of NN intervals for all 5-min segments [SDANN]) were low at nighttime and increased in the morning hours. There was a significant decline of 24-hour values of all HRV parameters (P < 0.001) and a strong negative correlation (P < 0.001) with increasing age. Mean 24-hour RR interval (P < 0.001), SDNN, mean SD of NN intervals for all 5-minute intervals (SDNNi), and SDANN (all P < 0.01) were significantly higher in men. Younger men also exhibited significantly higher values for vagus-associated parameters (root mean square successive difference [rMSSD], P < 0.05; SDNNi, P < 0.01); however, gender differences diminished with increasing age. CONCLUSION: Normal aging is associated with a constant decline of cardiac vagal modulation due to a significant decrease of nocturnal parasympathetic activity. The significant gender-related difference of HRV decreases with aging. These findings emphasize the need to determine age-, gender , and nycthemeral-dependent normal ranges for HRV assessment. PMID- 12890037 TI - Heart rate variability: back to the beginning. PMID- 12890039 TI - Specialized conduction cells in human pulmonary veins: fact and controversy. PMID- 12890038 TI - Evidence of specialized conduction cells in human pulmonary veins of patients with atrial fibrillation. AB - Specialized Conducting Cells in Human PV. INTRODUCTION: Depolarizations similar to those from the sinus node have been documented from the pulmonary veins after isolation procedures. We assessed the hypothesis that sinus node-like tissue is present in the pulmonary veins of humans. METHODS AND RESULTS: Pulmonary vein tissue was obtained from five autopsies (four individuals with a history of atrial fibrillation and one without a history of atrial arrhythmias) and five transplant heart donors. Autopsy veins were fixed in formaldehyde and processed for light microscopy to identify areas having possible conductive-like tissue. Areas requiring additional study were extracted from paraffin blocks and reprocessed for electron microscopy. Donor specimens were fixed in formaldehyde for histologic sections and glutaraldehyde for electron microscopy. Myocardial cells with pale cytoplasm were identified by light microscopy in 4 of the 5 autopsy subjects. Electron microscopy confirmed the presence of P cells, transitional cells, and Purkinje cells in the pulmonary veins of these cases. CONCLUSION: Our report is the first to show the presence of P cells, transitional cells, and Purkinje cells in human pulmonary veins. Whether these cells are relevant in the genesis of atrial fibrillation requires further study. PMID- 12890040 TI - Psychosocial aspects of patient-activated atrial defibrillation. AB - Atrial Defibrillator. INTRODUCTION: The atrial defibrillator empowers patients to cardiovert themselves from atrial arrhythmias at a time that is socially and physically acceptable, thereby preventing hospitalization. The long-term psychosocial effects of repeated use of the patient-activated atrial defibrillator at home are unknown. METHODS AND RESULTS: Eighteen patients underwent placement of the Jewel AF atrial defibrillator for persistent atrial fibrillation only. All patients performed manually activated cardioversions at home under self-administered sedation. Automatic shock therapies were disabled. Hospital Anxiety and Depression Scale and Multidimensional Health Locus of Control questionnaires were obtained before implant. All patients completed questionnaires 1 year after device implant and at long-term follow-up. The spouse or partner of each patient was interviewed to identify positive and negative aspects of manual cardioversion at home. The baseline patient scores for both anxiety (5.7 +/- 2.7) and depression (3.4 +/- 2.3) fell within the predefined range of normality. At 1 year, there was no significant change in anxiety (4.9 +/ 3.7, P = 0.39) or depression (2.4 +/- 1.8, P = 0.06). At long-term follow-up (mean 28 months), a total of 377 patient-activated cardioversions were performed out of hospital (median 15 per patient). Scores for anxiety (6.0 +/- 4.0, P = 0.70) and depression (3.2 +/- 2.5, P = 0.68) remained unchanged. CONCLUSION: During long-term follow-up, patient-activated cardioversion using the atrial defibrillator was not associated with increased anxiety or depression. The procedure was well tolerated by patients and their partners, offering an acceptable treatment option for patients with recurrent persistent atrial fibrillation. PMID- 12890041 TI - Use of atrial defibrillators to convert atrial fibrillation at home: psychosocial aspects. PMID- 12890042 TI - Prognostic significance of heart rate turbulence following ventricular premature beats in patients with idiopathic dilated cardiomyopathy. AB - Heart Rate Turbulence in Dilated Cardiomyopathy. INTRODUCTION: The aim of this study was to investigate the prognostic significance of heart rate turbulence (HRT) characterized by HRT onset and slope after ventricular premature beats in patients with idiopathic dilated cardiomyopathy (IDC). METHODS AND RESULTS: Blinded HRT analysis was performed in 242 patients with IDC who were enrolled in the Marburg Cardiomyopathy database between 1992 and 2000. During 41 +/- 23 months of follow-up, 54 patients (22%) died or underwent heart transplant. On Cox univariate regression analysis, abnormal HRT onset, HRT slope, HRT onset combined with HRT slope, left ventricular (LV) ejection fraction, LV size, and New York Heart Association (NYHA) functional class III showed a significant association with total mortality or the need for heart transplant. On multivariate analysis, abnormal HRT onset identified patients without transplant-free survival, as did LV size and NYHA class III heart failure. Major arrhythmic events were observed in 42 patients (17%) during follow-up. On univariate analysis, abnormal HRT onset, HRT onset combined with HRT slope, male sex, NYHA class III, LV ejection fraction, and LV size were associated with a higher incidence of major arrhythmic events. On multivariate analysis, only LV ejection fraction remained as a significant arrhythmia risk predictor, with a relative risk of 2.2 per 10% decrease in ejection fraction (95% confidence interval 1.5-3.2). CONCLUSION: In this selected patient population with IDC, HRT onset is a significant predictor of transplant-free survival, as are LV size and NYHA class. For arrhythmia risk stratification, however, only LV ejection fraction remained a significant risk predictor on multivariate analysis. PMID- 12890043 TI - Safety and efficacy of cryoablation of accessory pathways adjacent to the normal conduction system. AB - Cryoablation of Septal Accessory Pathways. INTRODUCTION: Catheter ablation has become a routine treatment for patients with Wolff-Parkinson-White syndrome because of its low risk and high efficacy; however, radiofrequency ablation in the septum close to the AV node or His bundle still carries a definite risk for AV block. Cryoenergy catheter ablation has recently become available. This technique has specific features, such as the ability to create reversible loss of function to predict the effects of ablation (ice mapping) and the adherence of the catheter tip to the endocardium with freezing, which avoids the risk for dislodgment. Both of these characteristics may minimize the risk of complications. The aim of this study was to analyze the effectiveness and safety of catheter cryoablation in 20 patients with para-Hisian or midseptal accessory pathways (AP). METHODS AND RESULTS: Eleven patients with para-Hisian and 9 patients with midseptal AP underwent catheter cryoablation. Ice mapping at -30 degrees C was performed to ascertain the disappearance of AP conduction and the absence of impairment of AV nodal conduction. If the expected result was obtained, cryoablation was performed by lowering the temperature to -75 degrees C for 4 minutes in order to create a permanent lesion. Cryoablation was successful in all patients using a mean of 1.2 +/- 0.4 applications. Recurrences occurred in 4 patients (20%) who underwent a second successful cryoablation session. No complications were observed. CONCLUSION: Cryoablation appears to be a safe and effective technique for ablation of APs close to the AV node or His bundle because of the ability to predict the acute effects of ablation with ice mapping before creation of an irreversible lesion. PMID- 12890044 TI - Cryoablation for septal accessory pathways: has the next ice age arrived? PMID- 12890045 TI - Defibrillation causes immediate cardiac dilation in humans. AB - Defibrillation Causes Dilation. INTRODUCTION: Prior studies in isolated heart tissue have shown both excitation and deexcitation to be the primary mechanism of defibrillation. This article presents the first evidence in man of deexcitation immediately following defibrillation by tracking the heart's mechanical response. METHODS AND RESULTS: The geometric changes of the ventricular chambers were measured before and after defibrillation in seven human subjects receiving an implantable cardioverter defibrillator (ICD). The ICD was used to produce approximately three episodes of ventricular fibrillation and defibrillation in each subject. Twenty-two two-dimensional echocardiographic images of the right ventricle (RV) and 11 images of the left ventricle (LV) were recorded and analyzed at 30 frames per second. Just over 2 seconds of each episode were digitized, beginning half a second before the defibrillation shock. Individual frames were analyzed to yield cross-sectional, ventricular chamber area as a function of time. Immediately following defibrillation, ventricular chambers dilated with significant fractional area increase (RV: 1.58 +/- 0.25, LV: 1.10 +/ 0.06), with peak dilation at 194 +/- 114 msec. CONCLUSION: Defibrillation causes a rapid increase in ventricular chamber area due to relaxation of the myocardium, suggesting that defibrillation synchronizes the cardiac cells to the deexcited state in man. PMID- 12890046 TI - Defibrillation efficacy and pain perception of two biphasic waveforms for internal cardioversion of atrial fibrillation. AB - Efficacy and Pain Perception of Two Biphasic Waveforms. INTRODUCTION: We evaluated the influence of the peak voltage of waveforms used for internal cardioversion of atrial fibrillation on defibrillation efficacy and pain perception. A low peak voltage biphasic waveform generated by a 500-microF capacitor with 40% tilt was compared to a standard biphasic waveform generated by a 60-microF capacitor with 80% tilt. METHODS AND RESULTS: In 19 patients with paroxysmal atrial fibrillation (79% male, age 55 +/- 11 years, 21% with heart disease), the atrial defibrillation threshold (ADFT) was determined during deep sedation with midazolam for both waveforms in a randomized fashion using a step up protocol. Internal cardioversion with a single lead (shock vector: coronary sinus to right atrium) was successful in 18 (95%) of 19 patients. ADFT energy and peak voltage were significantly lower for the low-voltage waveform (2.1 +/- 2.4 J vs 3.5 +/- 3.9 J, P < 0.01; 100 +/- 53 V vs 290 +/- 149 V, P < 0.01). Sedation then was reversed with flumazenil after ADFT testing. Two shocks at the ADFT (or a 3-J shock if ADFT >3 J) were administered to the patient using each waveform in random order. Pain perception was assessed using both a visual scale and a numerical score. ADFTs were above the pain threshold in 17 (94%) of 18 patients, even though the ADFT with the 500-microF waveform was <100 V in 63% of the patients. Pain perception was comparable for both waveforms (numerical score: 6.5 +/- 2.4 vs 6.3 +/- 2.6; visual scale: 5.4 +/- 2.6 vs 5.2 +/- 3.1; P = NS, 500 microF vs 60-microF). The second shock was perceived as more painful in 88% of the patients, independent of the waveform used. CONCLUSION: Despite a 66% lower peak voltage and a 40% lower energy, the 40% tilt, 500-microF capacitor biphasic waveform did not change the pain perceived by the patient during delivery of internal cardioversion shocks. Pain perception for internal cardioversion probably is not influenced by peak voltage alone and increases with the number of applied shocks. PMID- 12890048 TI - Transplantation of stem cells to the heart:. PMID- 12890047 TI - Mesenchymal stem cell injection induces cardiac nerve sprouting and increased tenascin expression in a Swine model of myocardial infarction. AB - Stem Cell Induces Cardiac Nerve Sprouting. INTRODUCTION: Mesenchymal stem cell (MSC) transplantation is a promising technique to improve cardiac function. Whether MSC can increase cardiac nerve density and contribute to the improved cardiac function is unclear. METHODS AND RESULTS: Anterior wall myocardial infarction was created in 16 swine. One month later, 6 swine were given MSC and fresh bone marrow (BM) into infarcted myocardium (MSC group). Four swine were given fresh BM only (BM group), and 6 swine were given culture media (MI-only group). The swine were sacrificed 95.8 +/- 3.5 days after MI. Six normal swine were used as control. Immunocytochemical staining was performed using antibodies against growth-associated protein 43 (GAP43), tyrosine hydroxylase (TH), and three subtypes of tenascin (R, C, and X). Five fields per slide were counted for nerve density. The results show the following. (1). There were more GAP43 positive nerves in the MSC group than in the BM, MI-only, or Control group (P < 0.0001). TH staining showed higher nerve densities in the MSC group than in the MI-only (P < 0.01) or Control group (P < 0.0001) in the atria. (2). There were more sympathetic (TH-positive) nerves in myocardium distant from infarct than in the peri-infarct area (P < 0.05). (3). Optical intensity and color analyses showed significantly higher tenascin R and tenascin C expression in the MSC and BM groups than in the MI-only or Control group (P < 0.01). CONCLUSION: MSC injected with BM into swine infarct results in overexpression of cardiac tenascin, increased the magnitude of cardiac nerve sprouting in both atria and ventricles, and increased the magnitude of atrial sympathetic hyperinnervation 2 months after injection. PMID- 12890049 TI - Fiber orientation and cell-cell coupling influence ventricular fibrillation dynamics. AB - Cell Coupling Influences VF Dynamics. INTRODUCTION: The structure of ventricular fibrillation (VF) is influenced by regional differences in action potential durations and perhaps restitution kinetics and fiber anisotropy. The spatial organization of VF was investigated by measuring the cross-correlation (CC) and mutual information (MI) of membrane potential (Vm) oscillations recorded from multiple sites. METHODS AND RESULTS: Rabbit hearts (n = 6) were retrogradely perfused and stained with di-4-ANEPPS, and VF was elicited by burst pacing. Vm oscillations were recorded optically from multiple locations on the epicardium using a 16 x 16 photodiode array or a 72 x 78 CCD camera. The spatial organization of VF was investigated by calculating the maximum CC (CCmax) and MI (MImax) that can be obtained between any two sites. CCmax and MImax were extended to all pixels and served as indices of the similarities between Vm transients at a reference pixel and all other pixels on the map. We found that maps of CCmax and MImax did not contain discrete regions with high CC or MI. However, CCmax and MImax decreased monotonically with increasing distance between any arbitrarily chosen reference pixel and all other pixels. In VF, maps of CCmax and MImax revealed elliptical gradients of CC and MI that were closely aligned with fiber orientation, with major axis at 127 degrees +/- 8 degrees on the left ventricles. CONCLUSION: CC and MI analysis in fibrillation provides new evidence that anisotropy of fiber orientation and cell-cell coupling have a direct influence on VF dynamics. PMID- 12890050 TI - Combined amiodarone and silymarin treatment, but not amiodarone alone, prevents sustained atrial flutter in dogs. AB - Amiodarone/Silymarin Treatment for Sustained Atrial Flutter. INTRODUCTION: Because amiodarone generates free radicals that may mediate amiodarone's toxicity, simultaneous therapy with an antioxidant might be beneficial if the antioxidant did not impair amiodarone's antiarrhythmic action. We tested whether simultaneous administration of a flavonoid antioxidant, silymarin, altered the electrophysiologic (EP) actions of amiodarone in 62 open chest dogs with electrically induced atrial flutter created by a Y-shaped right atrial incision. METHODS AND RESULTS: Fifteen dogs received oral amiodarone (600 mg/day); 15 dogs received amiodarone (600 mg/day) and silymarin (70 mg bid); and 8 dogs received silymarin (70 mg bid) alone. All dosing was for 8 weeks; 24 control dogs received no drugs prior to induction of atrial flutter. Atrial flutter was induced by rapid right atrial pacing, and EP measurements were made before (presurgical) and after (postsurgical) creation of a Y-shaped right atrial incision. There was no difference in the frequency of induction of atrial flutter lasting >30 minutes among amiodarone-treated (8/15 [53%]), silymarin-treated (4/6 [67%]), and control (15/21 [71%]) groups, whereas the frequency of induction in the amiodarone+silymarin dogs (2/15 [13%]) was significantly reduced (P = 0.008) compared with the other three groups. Both amiodarone and amiodarone+silymarin treatment prolonged the presurgical and postsurgical right atrial effective refractory period (P = 0.012) compared with control; however, there was no significant difference in either parameter between the amiodarone+silymarin treated and amiodarone-treated groups. The increase in atrial flutter mean cycle length (postsurgical minus presurgical) was significantly (P = 0.005) less in the amiodarone+silymarin-treated and control dogs compared with the amiodarone treated dogs (16 +/- 11 msec for amiodarone+silymarin; 24 +/- 8 msec for control; and 42 +/- 14 msec for amiodarone treatment). Amiodarone+silymarin treatment resulted in a longer postsurgical right atrial refractory period (155 +/- 13 msec) than atrial flutter mean cycle length (154 +/- 19 msec), consistent with reduction and/or elimination of the excitable gap. Silymarin alone did not exert significant EP or antiarrhythmic action. CONCLUSION: Amiodarone exerted no preventative antiarrhythmic action in this atrial flutter model, probably because it could not reduce the excitable gap of atrial flutter. However, an antioxidant, silymarin, without a direct antiarrhythmic action, when administered together with amiodarone, potentiated amiodarone's antiarrhythmic actions and prevented sustained atrial flutter by reduction and/or elimination of the excitable gap. PMID- 12890051 TI - Biphasic versus monophasic cardioversion in shock-resistant atrial fibrillation:. AB - Biphasic versus Monophasic Cardioversion. INTRODUCTION: Cardioversion of atrial fibrillation using monophasic transthoracic shocks occasionally is ineffective. Biphasic cardioversion requires less energy than monophasic cardioversion, but its efficacy in shock-resistant atrial fibrillation is unknown. Thus, we compared the efficacy of cardioversion using biphasic versus monophasic waveform shocks in patients with atrial fibrillation previously refractory to monophasic cardioversion. METHODS AND RESULTS: Fifty-six patients with prior failed monophasic cardioversion were randomized to either a 360-J monophasic damped sinusoidal shock or biphasic truncated exponential shocks at 150 J, followed by 200 J and then 360 J, if necessary. If either waveform failed, patients were crossed over to the other waveform. The primary endpoint was defined as the proportion of patients achieving sinus rhythm following initial randomized therapy. Stepwise multivariate logistic regression examined independent predictors of shock success, including patient age, sex, left atrial diameter, body mass index, drug therapy, and waveform. Twenty-eight patients were randomized to the biphasic shocks and 28 to the monophasic shocks. Sinus rhythm was restored in 61% of patients with biphasic versus 18% with monophasic shocks (P = 0.001). Seventy-eight percent success was achieved in patients who crossed over to the biphasic shock after failing monophasic cardioversion, whereas only 33% were successfully cardioverted with a monophasic shock after crossover from biphasic shock (P = 0.02). Overall, 69% of patients who received a biphasic shock at any point in the protocol were cardioverted successfully, compared to 21% with the monophasic shock (P < 0.0001). The type of shock was the strongest predictor of shock success (P = 0.0001) in multivariate logistic regression. CONCLUSION: An ascending sequence of 150-, 200-, and 360-J transthoracic biphasic cardioversion shocks are successful more often than a single 360-J monophasic shock. Thus, biphasic shocks should be the recommended configuration of choice for all cardioversions. PMID- 12890052 TI - Heterogeneous regional endocardial repolarization is associated with increased risk for ischemia-dependent ventricular fibrillation after myocardial infarction. AB - Repolarization Heterogeneity and Sudden Death Risk. INTRODUCTION: The aim of this study was to investigate whether the characteristics of endocardial ventricular repolarization are associated with differential risk for sudden death. Prolonged surface QT interval is associated with increased arrhythmic risk after myocardial infarction (MI), but the underlying mechanism of QT prolongation and its relation to lethal arrhythmias are unclear. METHODS AND RESULTS: Ventricular fibrillation (VF) risk was assessed in 12 dogs 1 month after anterior MI during an exercise test coupled with brief circumflex coronary occlusion. Susceptible dogs (n = 5) developed VF during the brief ischemic episode, whereas resistant dogs did not (n = 7). Surface QT interval was measured at rest. Endocardial electroanatomic catheter maps of left ventricular repolarization were obtained in four unique regions identified by echocardiography and compared between groups. Compared to resistant dogs, susceptible dogs were characterized by prolonged surface QT intervals (240 +/- 10 msec vs 222 +/- 7 msec, P = 0.04). In addition, they had lower baroreflex sensitivity (9.7 +/- 1.5 msec/mmHg vs 28 +/- 9.8 msec/mmHg, P < 0.01) and a tachycardic response to acute ischemia suggesting higher propensity for stronger sympathetic reflexes. Surface QT interval prolongation in susceptible dogs was due to a marked heterogeneity of endocardial left ventricular repolarization (239 +/- 42 msec, basal anterior wall vs 197 +/- 35, lateral wall; P < 0.001). Resistant animals had no regional differences in endocardial repolarization. CONCLUSION: Sympathetic activation following MI not only produces adverse structural remodeling but also contributes to adverse electrophysiologic remodeling resulting in heterogeneous ventricular repolarization and in a myocardial substrate conducive to lethal reentrant arrhythmias. PMID- 12890053 TI - Antiarrhythmic effects of JTV-519, a novel cardioprotective drug, on atrial fibrillation/flutter in a canine sterile pericarditis model. AB - Effect of JTV-519 on AF. INTRODUCTION: A new cardioprotective drug, JTV-519, blocks Na+ current and inwardly rectifying K+ current and inhibits Ca2+ current. However, its role in atrial electrophysiology is unknown. We investigated the antiarrhythmic effects of JTV-519 on atrial fibrillation/flutter in the canine sterile pericarditis model. METHODS AND RESULTS: In nine dogs with sterile pericarditis, 38 episodes of sustained (>30 sec) atrial fibrillation (8 dogs) and 24 episodes of sustained atrial flutter (7 dogs) were induced by rapid atrial pacing. When atrial fibrillation or atrial flutter was sustained >15 minutes, it was cardioverted and reinduced. The inducibility of atrial fibrillation/flutter, the atrial effective refractory period, and the intra-atrial conduction time were compared before and after the continuous infusion of JTV-519 (0.03 mg/kg/min). JTV-519 significantly decreased the mean number of sustained atrial fibrillation episodes (from 4.2 +/- 2.9 to 0 +/- 0, P < 0.01). In contrast, atrial flutter was still inducible in 4 dogs after JTV-519 (from 2.7 +/- 2.5 to 1.6 +/- 2.1, P = NS). JTV-519 significantly prolonged effective refractory period (from 123 +/- 18 to 143 +/- 14 msec, from 127 +/- 18 to 151 +/- 12 msec, and from 132 +/- 13 to 159 +/- 9 msec at basic cycle lengths of 200, 300, and 400 msec, respectively, P < 0.01), but it did not affect the intra-atrial conduction time (from 47 +/- 11 msec to 48 +/- 11 msec, P = NS). CONCLUSION: JTV-519 had significant protective effects on atrial fibrillation in the canine sterile pericarditis model, mainly by increasing effective refractory period, suggesting that it may have potential as a novel antiarrhythmic agent for atrial fibrillation. PMID- 12890054 TI - Effects of amiodarone and dronedarone on voltage-dependent sodium current in human cardiomyocytes. AB - Effect of Dronedarone on Cardiac Na Current. INTRODUCTION: Amiodarone (AM) is a highly effective antiarrhythmic agent used in the management of both atrial and ventricular arrhythmias. Its noniodinated analogue dronedarone (SR) may have fewer side effects than AM. In this study, we compared the effects of AM and SR on the sodium current I(Na) in human atrial myocytes. METHODS AND RESULTS: INa was studied with the whole-cell, patch clamp technique. Both AM and SR induced a dose-dependent inhibition of I(Na) recorded at -40 mV from a holding potential of -100 mV. AM inhibited I(Na) by 41%+/- 11% (n = 4) at 3 microM, and by 80%+/- 7% (n = 5) at 30 microM. SR produced more potent block, inhibiting INa significantly at only 0.3 microM (23%+/- 10%, n = 4) and completely (97%+/- 4%, n = 4) at 3 microM. Both AM and SR had only moderate effects on voltage-dependent properties of I(Na) (current-voltage relationship, availability for activation) and had no effect on the current decay kinetics. CONCLUSION: Both AM and SR inhibit I(Na) significantly in single human atrial cells, showing that the two drugs have Class I antiarrhythmic properties. The acute effects of SR are more potent than those of AM. The study supports the idea that the iodinated form of the molecule has no part in the acute effect of AM on Na+ channels. PMID- 12890055 TI - Spontaneous premature complexes during a narrow QRS tachycardia: what is the mechanism? PMID- 12890056 TI - Tachycardia during bradycardia: what is the mechanism? PMID- 12890057 TI - Three-dimensional right atrial geometry construction and catheter tracking using cutaneous patches. PMID- 12890059 TI - Reducing atrial defribrillation thresholds in patients with persistent atrial fibrillation. PMID- 12890061 TI - Variable patterns of septal activation in patients with left bundle branch block and heart failure. PMID- 12890063 TI - Site-selective atrial septal puncture. PMID- 12890064 TI - Mechanisms of clinical arrhythmias. PMID- 12890066 TI - Exploring sex differences in the relationship between depressive symptoms and dementia incidence: prospective results from the PAQUID Study. AB - OBJECTIVES: To estimate the predictive relationship between depressive symptoms and 8-year dementia incidence in a large prospective community sample of French older adults and to compare the effect magnitude for men and women. DESIGN: Prospective population-based cohort with four interview waves and complete vital status ascertainment. SETTING: Urban and rural communities in the Aquitaine Region (Gironde and Dordogne), southwest France. PARTICIPANTS: Three thousand seven hundred seventy-seven adults aged 65 and older residing in noninstitutional settings at study baseline. MEASUREMENTS: Each participant was interviewed by a neuropsychologist and screened for dementia with the Mini-Mental State Examination, a cognitive test battery, and a standardized questionnaire designed to ascertain the presence of the criteria for dementia according to the Diagnostic and Statistical Manual for Mental Disorders, Third Edition, Revised (DSM-III-R). Dementia status and subtype were confirmed using neurological examination and categorized according to DSM-III-R criteria for dementia and the National Institute of Neurological Disorders and Stroke/Alzheimer's and Related Disorders Association criteria. The Hachinski score was calculated to specify the etiology: possible or probable Alzheimer's disease, vascular dementia, and other types of dementia. Depressive symptomatology was evaluated using the Center for Epidemiologic Studies-Depression scale. Statistical analyses were weighted to correct for attrition not due to mortality. RESULTS: Ninety-seven men (incidence rate: 14.4/1,000) and 183 women (Incidence rate: 19.0/1,000) developed dementia during 8 years of follow-up. Baseline prevalence of depressive symptomatology was 12.9% for men and 14.7% for women. Depressive symptoms increased risk of dementia at subsequent interview wave, but only for men (odds ratio (OR) (men) = 3.5, 95% confidence interval (CI) = 1.9-6.5; OR (women) = 1.2, 95% CI = 0.7-2.0, P-value for sex difference = 0.03). The hypothesis that vascular depression might explain the observed sex difference was studied, and it was found that risk was 50% higher for men with hypertension who were depressed than for normotensive men. For women, hypertension status did not modify the absence of an association. CONCLUSION: This study supports the hypothesis of a relationship between proximal depressive symptomatology and dementia in men, but distant depression did not increase dementia risk in this sample. The results suggest that depression in older men might reflect a form of vascular depression associated with cerebral vascular pathology or multiinfarct disease that may amplify the dementing or declining process, hence accelerating the onset of manifest symptoms of dementia. PMID- 12890067 TI - Mild cognitive deterioration with subcortical features: prevalence, clinical characteristics, and association with cardiovascular risk factors in community dwelling older persons (The InCHIANTI Study). AB - OBJECTIVES: To identify subjects with minimal or mild nonamnestic cognitive impairment with parkinsonian stance and gait and investigate vascular correlates of this condition. DESIGN: First wave of an epidemiological longitudinal study (InCHIANTI) on factors predicting loss of mobility in older persons. SETTING: The Chianti geographic area (Tuscany, Italy). PARTICIPANTS: Five hundred fifty-six subjects aged 70 to 90 with Mini-Mental State Examination (MMSE) scores greater than 23 of 30, of the 1,260 persons aged 65 and older randomly selected from the population registry of Greve in Chianti and Bagno a Ripoli, two small towns near Florence. METHODS: Low cognitive performance (LCP) was defined as an age- and education-adjusted MMSE below the 50th percentile for the InCHIANTI population. Subcortical features were plastic rigidity on neurological examination (parkinsonism), gait disturbance (small-step gait or parkinsonian gait), and dysexecutive features. Two hundred forty-three participants had high cognitive performance, 166 had LCP without subcortical features, and 75 had LCP with subcortical features. Vascular risk factors were hypertension, atrial fibrillation or pathological findings on electrocardiogram (ECG), low serum high density lipoprotein (HDL) or high low-density lipoprotein cholesterol, diabetes mellitus, obesity, and heavy smoking. RESULTS: Three main vascular risk factors were significantly more prevalent in LCP with subcortical features: hypertension (P =.02), atrial fibrillation or ECG changes (P =.04), and low HDL cholesterol (P =.001). LCP with subcortical features was significantly associated with cerebrovascular risk factors (P =.001). CONCLUSION: Gait disturbance and nonamnestic cognitive symptoms might be the consequence of subcortical vascular damage. PMID- 12890068 TI - Is large-scale community memory screening feasible? Experience from a regional memory-screening day. AB - OBJECTIVES: To investigate whether a large-scale memory-screening program for community-dwelling elders would be successful in identifying individuals with a high probability of dementia in need of further assessment that would result in the earlier diagnosis of dementia. DESIGN: A descriptive study of experience with a volunteer sample. SETTING: Ten sites (e.g., senior centers, churches, clinics) throughout New England on October 29, 1999. PARTICIPANTS: Trained volunteer clinicians evaluated 497 community-dwelling individuals on the screening day. An additional 162 subjects who could not be accommodated on that day were subsequently screened at local sites by appointment during the following month. MEASUREMENTS: Subjects participated in a standardized format consisting of an educational lecture, followed by individual screenings with the 7-minute screen (7MS) with locally trained staff. Subjects were informed immediately of test results and counseled regarding follow-up options. A survey was conducted with these subjects and their primary care physicians over the following year. RESULTS: Because the groups tested at different times were not statistically different in terms of demographics, they were combined in the analysis. One hundred ten (16.7% of all screened) individuals received high/retest scores on the 7MS. They were advised to seek diagnostic evaluation and encouraged to have results sent to their primary care physicians (PCPs). Of those followed up, 64% reported that they followed up the screening results with their PCP. More than one-third (38%) of participants with a high/retest score on the 7MS had inconclusive findings on follow-up or were awaiting further diagnostic evaluation. Of those for whom follow-up data were available, 10 (9%) were diagnosed with probable Alzheimer's disease (AD), and an additional nine (8%) who had previous diagnoses of AD were correctly identified by the 7MS. Anecdotally, feedback from participants indicated a high level of satisfaction with the process. Participants reported that the educational talk and the possibility of early detection were the most helpful components of the screening program. Moreover, most individuals surveyed in follow-up would recommend the program to a friend or family member. CONCLUSION: A follow-up survey of participants and their physicians supported the conclusion that a community memory-screening program might detect individuals who were previously unknown to have cognitive problems. Furthermore, such a program was highly acceptable to participants. The small number of individuals diagnosed with dementia as a result of the screening program indicates that this form of screening may be inefficient as performed. Multiple obstacles to seeking follow-up care were identified and would need to be addressed in larger-scale programs to make this a worthwhile endeavor. The experience gained in this memory screening program might aid in the planning of better programs, which will be essential if early diagnosis is to keep pace with the growth of treatments for dementia. PMID- 12890069 TI - Organizational characteristics and restraint use for hospitalized nursing home residents. AB - OBJECTIVES: To examine the effect of organizational characteristics on physical restraint use for hospitalized nursing home residents. DESIGN: Secondary analysis of data obtained between 1994 to 1997 in a prospective phase lag design experiment using an advanced practice nurse (APN) intervention aimed at reducing physical restraint for a group of hospitalized nursing home residents. SETTING: Eleven medical and surgical units in one 600-bed teaching hospital. PARTICIPANTS: One hundred seventy-four nursing home residents aged 61 to 100, hospitalized for a total of 1,085 days. MEASUREMENTS: Physical restraint use, APN intervention, age, perceived fall risk, behavioral phenomena, perceived treatment interference, mental state, severity of illness, day of week, patient-registered nurse (RN) ratio, patient-total nursing staff ratio, and skill mix. RESULTS: Controlling for the APN intervention, age, and patient behavioral characteristics (all of which increased the likelihood of restraint use), weekend days as an organizational characteristic significantly increased the odds of restraint (weekend day and patient-RN ratio on physical restraint use: odds ratio (OR) = 1.92, 95% confidence interval (CI) = 1.38-2.68, P <.001; weekend day and patient-total staff ratio on physical restraint use: OR = 1.91, 95% CI = 1.37-2.66, P <.001; weekend day and skill mix on physical restraint use: OR = 1.91, 95% CI = 1.37 2.67, P <.001). CONCLUSION: Key findings suggest that organization of hospital care on weekends and patient characteristics that affect communication ability, such as severely impaired mental state, English as a second language, sedation, and sensory-perceptual losses, may be overlooked variables in restraint use. PMID- 12890070 TI - Age-related changes of circadian rhythms and sleep-wake cycles. AB - OBJECTIVES: To compare relationships between the sleep-wake cycle and endogenous circadian rhythms in young and older adults and to examine correlates between evening naps and circadian rhythms in older adults. DESIGN: For 1 week of home recording, subjects wore wrist-activity monitors and kept daily sleep logs. After the home monitoring, subjects entered the laboratory on a 90-minute sleep-wake schedule and were monitored on this schedule for at least 30 hours. SETTING: Community living and laboratory. PARTICIPANTS: Sixty-seven young adults, aged 18 to 32, and 56 older adults, aged 60 to 75, who were healthy and had few sleep complaints. MEASUREMENTS: Times of nocturnal sleep, out-of-bed napping, and illumination were obtained at home. Sleep propensity and oral body temperature (OBT) were measured in the laboratory, along with circadian rhythms of cortisol and 6-sulfatoxymelatonin (aMT6s, assayed from urine samples collected every 90 minutes). RESULTS: Home sleep times and illumination acrophases (fitted peak times) were advanced in older adults. The phase angles (time intervals) between onset of aMT6s and sleep onset were not changed in older adults, but sleep offset was more advanced than acrophase and offset of aMT6s with aging. Acrophases of cortisol and sleep propensity were advanced in older adults to the same extent as sleep times, but OBT was less advanced than sleep times. Older adults who took evening naps showed more advanced sleep offset and circadian rhythms of aMT6s, but there were no differences in the phase angles of sleep onset and circadian rhythms of aMT6s and cortisol compared with older adults who did not take evening naps. CONCLUSION: Measuring different circadian markers suggested different phase relationships between the sleep-wake cycle and endogenous circadian rhythms in aging. Early awakening in older adults cannot be explained simply by a relative phase advance of the circadian system. Evening naps and advanced illumination may play a role in the advance of the circadian system in aging. PMID- 12890071 TI - The relationship between psychological factors and disabling musculoskeletal pain in community-dwelling older persons. AB - OBJECTIVES: To determine the relationship between two psychological factors (depressive symptoms and low functional self-efficacy) and the occurrence of disabling musculoskeletal pain in community-dwelling older persons. DESIGN: A 12 month prospective cohort study. SETTING: Community-based. PARTICIPANTS: Two hundred twenty-six community-dwelling persons residing in the greater New Haven, Connecticut, region, aged 70 and older, who had a history of clinically evident musculoskeletal pain and were independent in bathing, walking, dressing, and transferring. MEASUREMENTS: Levels of depressive symptoms and functional self efficacy were determined during a comprehensive baseline assessment along with information regarding participants' demographic, medical, and physical/cognitive status. The occurrence of disabling musculoskeletal pain, defined as staying in bed for at least one-half day or cutting down on one's usual activities due to joint or back pain, was ascertained during monthly interviews. RESULTS: The mean number of months of disabling musculoskeletal pain, adjusted for baseline covariates, increased from the lowest to the highest quartile of depressive symptoms: 1.2 (95% confidence intervals = 0.8-1.7), 1.4 (1.0-2.0), 2.0 (1.5-2.8), 2.3 (1.7-3.1), respectively, P for trend =.002. The corresponding results for functional self-efficacy were (from highest to lowest quartile) 1.4 (1.0-2.0), 1.6 (1.2-2.2), 1.6 (1.2-2.2), 2.2 (1.6-3.0), P for trend =.068. There was no interaction between depressive symptoms and functional self-efficacy. CONCLUSION: Depressive symptoms and, to a lesser extent, low functional self-efficacy were each associated with the occurrence of disabling musculoskeletal pain among community-dwelling older persons. PMID- 12890072 TI - Propoxyphene use by community-dwelling and institutionalized elderly Medicare beneficiaries. AB - OBJECTIVES: To provide the first comparable national prevalence estimates on use of propoxyphene, a potentially inappropriate drug, by elderly Medicare beneficiaries living in the community and institutions and to determine whether institutionalized beneficiaries are at a greater risk for receiving propoxyphene than community-dwelling beneficiaries. DESIGN: Cross-sectional study. SETTING: U.S. representative sample of elderly using Medicare database. PARTICIPANTS: Nationally representative sample of community-dwelling (n = 9,851, weighted n = 32.5 million) and institutionalized (n = 1,099, weighted n = 2.3 million) Medicare beneficiaries aged 65 and older. MEASUREMENTS: National estimates on prevalence of propoxyphene use and the odds of receiving propoxyphene were the two main outcome measures. RESULTS: Annual prevalence of propoxyphene use in 1998 was 6.8% by all community-dwelling elderly beneficiaries and 15.5% by institutionalized elderly beneficiaries. Beneficiaries in long-term care facilities had almost 40% higher odds of receiving propoxyphene (odds ratio = 1.38, 95% confidence interval = 1.1-1.8) than beneficiaries in the community even after controlling for other factors in a logistic regression. Other risk factors include female, rural residence, poor health, and history of osteoporosis or hip fracture. Beneficiaries residing in regions in the midwest and south were more than twice as likely to receive propoxyphene as those in the mid-Atlantic area. CONCLUSION: These results show that propoxyphene use by U.S. community-dwelling seniors is high but is much higher in the institutionalized population. These findings suggest that prescribing for older adults with pain could be improved, especially for vulnerable long-term care residents. PMID- 12890073 TI - Late-life depression is associated with arterial stiffness: a population-based study. AB - OBJECTIVES: To determine whether arterial stiffness is associated with depression in the elderly. DESIGN: Population-based cross-sectional study. SETTING: In Ommoord, a suburb of Rotterdam, the Netherlands. PARTICIPANTS: Three thousand seven hundred four subjects of the Rotterdam Study aged 60 and older. MEASUREMENTS: Arterial stiffness was assessed using the distensibility of the carotid artery and the carotid-femoral pulse wave velocity. All participants were screened for depressive symptoms with the Center for Epidemiologic Studies Depression scale. Those with depressive symptoms had a psychiatric evaluation to establish a diagnosis of depressive disorders according to Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition (DSM-IV) criteria. RESULTS: Participants with increased arterial stiffness were more likely to have depressive symptoms. Odds ratios (ORs) for depressive symptoms were 1.24 (95% confidence interval (CI) = 1.01-1.52) per standard deviation decrease in carotid distensibility and 1.17 (95% CI = 1.00-1.38) per standard deviation increase in aortic pulse wave velocity. The association was stronger for depressive disorders meeting DSM-IV criteria (OR = 1.44, 95% CI = 1.03-2.03; OR = 1.48, 95% CI = 1.16 1.90, respectively). Control for atherosclerosis, as measured by the ankle-to brachial index or presence of plaques in the carotid artery, did not change the associations. CONCLUSION: This study shows an association between arterial stiffness and depression in the elderly. The findings are compatible with the vascular depression hypothesis. These data suggest that arterial stiffness may partly cause the proposed relationship between vascular factors and depression. PMID- 12890074 TI - Predictive value of nonspecific symptoms for acute illness in nursing home residents. AB - OBJECTIVES: To examine the predictive value of nonspecific symptoms for acute illness in nursing home residents. DESIGN: Prospective, observational study. SETTING: Academic nursing home located in an urban setting. PARTICIPANTS: Two hundred two newly admitted residents. MEASUREMENTS: Eleven nonspecific symptoms were ascertained by review of observations documented by nursing home staff in the medical record. Research staff independently identified acute illness according to previously established criteria from nurse report and medical record review. Follow-up was divided into 10-day intervals, and concordance between nonspecific symptoms and acute illness within these intervals was determined. Predictive values were calculated according to standard formulae. RESULTS: Nonspecific symptoms and acute illnesses occurred in 21.7% and 12.5% of 10-day intervals, respectively. Positive predictive values (PPVs) were highest for lethargy, weakness, and decreased appetite, each of which correctly predicted the presence of an acute illness one out of two times the symptoms were reported. Agitation and disorientation predicted an acute illness one out of three times, and falls predicted an acute illness one out of four times. Overall, the PPV of the occurrence of any nonspecific symptom was 0.24, and the negative predictive value of the absence of nonspecific symptoms was 0.91. CONCLUSION: Hypoactive nonspecific symptoms are more likely than other nonspecific symptoms to be signs of incipient acute illness. Studies are needed to determine whether an intervention in residents with these nonspecific symptoms can enable earlier detection and treatment of acute illness. PMID- 12890075 TI - Tetanus immunity in long-term care facilities. AB - OBJECTIVES: To investigate the correlates of tetanus immunity in the elderly residing in a long-term care facility in Hawaii. DESIGN: Cross-sectional. SETTING: A nursing home in Honolulu, Hawaii. PARTICIPANTS: Sixty subjects aged 65 and older: 30 men and 30 women. MEASUREMENTS: The interview included demographic information, immunization history, military service information, and other potential risk factors for tetanus. Serum tetanus antibody titers were measured. RESULTS: The data showed that 76.7% (46/60) had adequate tetanus titers. This is in stark contrast to previous studies, which have reported immunity rates of 27% to 46% in similar populations. There were significant associations between immunity and prior history of military service. There were no significant associations between immunity and past history of immunization, education, socioeconomic status, or sex. CONCLUSION: History of immunization from patients, families, or medical charts may be unreliable indicators of tetanus immunity. Recognizing patterns of and barriers to immunization could have important consequences for public health policy in long-term care institutions. PMID- 12890076 TI - Sarcopenia, calf circumference, and physical function of elderly women: a cross sectional study. AB - OBJECTIVES: To determine whether calf circumference (CC), related to appendicular skeletal muscle mass, can be used as a measure of sarcopenia and is related to physical function. DESIGN: Retrospective analysis of data from 1992 to 1994 of the European Patient Information and Documentation Systems Study. SETTING: Community setting in France. PARTICIPANTS: One thousand four hundred fifty-eight French women aged 70 and older without previous history of hip fracture were recruited from the electoral lists. MEASUREMENTS: Muscular mass was assessed using dual-energy x-ray absorptiometry (DEXA). CC was measured using a tape measure. Anthropometric measurements (height; weight; and waist, hip, and calf circumference), strength markers (grip strength), and self-reported physical function were also determined. Sarcopenia was defined (using DEXA) as appendicular skeletal muscle mass (weight (kg)/height (m2)) less than two standard deviations below the mean of a young female reference group. RESULTS: The prevalence of sarcopenia was 9.5%. CC was correlated with appendicular skeletal muscle mass (r = 0.63). CC under 31 cm was the best clinical indicator of sarcopenia (sensitivity = 44.3%, specificity = 91.4%). CC under 31 cm was associated with disability and self-reported physical function but not sarcopenia (defined using DEXA), independent of age, comorbidity, obesity, income, health behavior, and visual impairment. CONCLUSION: CC cannot be used to predict sarcopenia defined using DEXA but provides valuable information on muscle-related disability and physical function. PMID- 12890077 TI - The associations between physical activity and inflammatory markers in high functioning older persons: MacArthur Studies of Successful Aging. AB - OBJECTIVES: To determine the relationships between recreational activity, house/yard work activity, work activity, and total physical activity and high levels of two peripheral blood markers of inflammation: interleukin-6 (IL-6) and C-reactive protein (CRP). DESIGN: Cross-sectional study. SETTING: Three communities (Durham, NC; New Haven, CT; and East Boston, MA). PARTICIPANTS: Eight hundred seventy persons aged 70 to 79 who were in the top third of community dwelling older persons with respect to physical and cognitive functioning. MEASUREMENTS: Blood levels of IL-6 and CRP and self-reported recreational activity, house/yard work activity, work activity, and total physical activity. RESULTS: The adjusted odds ratios (AORs) for individuals with high levels of recreational activity to have values in the top tertiles of IL-6 and CRP were 0.65 (95% confidence interval (CI) = 0.48-0.87) and 0.70 (95% CI = 0.51-0.95), respectively. The AORs for those with a high level of house/yard work activity to have values in the top tertile of IL-6 and CRP were 0.90 (95% CI = 0.67-1.20) and 0.70 (95% CI = 0.50-0.96), respectively. High levels of house/yard work and recreational activity were independently associated with lower risk of high CRP. CONCLUSION: The association between high levels of recreational activity and lower levels of the inflammatory markers IL-6 and CRP suggests a mechanism for its protective effect and supports interventions that increase physical activity in older persons. Such potential benefits of increased physical activity on inflammatory markers will need to be confirmed in clinical trials. PMID- 12890078 TI - Usefulness of a linear analog scale questionnaire to measure health-related quality of life in elderly patients with chronic obstructive pulmonary disease. AB - OBJECTIVES: To examine the validity, discriminatory ability, and responsiveness of health-related quality-of-life (HRQoL) questionnaires using a linear analog scale (Quality of Life (QOL) scale) for chronic obstructive pulmonary disease (COPD). DESIGN: Cross-sectional and longitudinal. SETTING: Outpatient. PARTICIPANTS: One hundred two elderly subjects with mild to severe COPD. MEASUREMENTS: Scores on the QOL scale, the St. George's Respiratory Questionnaire (SGRQ), and the 36-item Short-Form Health Survey questionnaire (SF-36) and various clinical parameters were recorded. The correlations between these QOL questionnaires and various clinical parameters were then examined. The responses of 31 elderly COPD patients to the QOL scale and the SGRQ before and 3 months after the completion of a comprehensive pulmonary rehabilitation program were compared longitudinally. RESULTS: On cross-sectional study, the QOL scale showed a significant correlation with the total score and three components of the SGRQ. The QOL scale correlated significantly with all components of the SF-36, but the total SGRQ score correlated with only six components of the SF-36, excluding vitality and the mental health index. Both the QOL scale and the total score of the SGRQ correlated significantly with the oxygen cost diagram (OCD), Morale scale, 6-minute walking distance (6MWD), forced expiratory volume in 1 second, and instrumental activities of daily living (IADL) score. When subjects were divided into three groups according to disease severity (mild, moderate, severe) using American Thoracic Society guidelines, the total SGRQ score discriminated between the three groups. The QOL scale could not discriminate between mild and moderate or moderate and severe. On longitudinal study, 3 months after finishing the comprehensive pulmonary rehabilitation program, the QOL scale, the SGRQ, 6MWD, and OCD all showed significant improvement. The difference in the QOL scale after the comprehensive pulmonary rehabilitation program showed a significant correlation with changes in the SGRQ total score and the OCD but not with the 6MWD. CONCLUSION: The QOL scale is similar to more-complex questionnaires such as the SGRQ in terms of validity and responsiveness for evaluating disease-specific HRQoL in elderly COPD patients. In clinical settings, the QOL scale, as a simple questionnaire, may be useful for disease-specific HRQoL assessments in elderly COPD patients. PMID- 12890079 TI - Metastatic prostate cancer: complications and treatment. AB - Despite highly successful treatments for localized prostate cancer, approximately 35% to 40% of men will eventually experience a detectable rise in serum prostate specific antigen. A portion of these men will go on to experience clinically expressed extracapsular disease, with as many as two-thirds having evidence of bone involvement. Diagnosis of skeletal involvement involves serum markers of disease progression and radiological evaluation. Skeletal-related events are numerous and include bone pain, spinal cord compression, vertebral collapse, and pathological fractures. Current treatments for advanced prostate cancer include individual or combined hormonal therapies, chemotherapy, radiotherapy, surgical treatments, and most recently, antiresorptive medications. PMID- 12890080 TI - Current status of metals as therapeutic targets in Alzheimer's disease. AB - There is accumulating evidence that interactions between beta-amyloid and copper, iron, and zinc are associated with the pathophysiology of Alzheimer's disease (AD). A significant dyshomeostasis of copper, iron, and zinc has been detected, and the mismanagement of these metals induces beta-amyloid precipitation and neurotoxicity. Chelating agents offer a potential therapeutic solution to the neurotoxicity induced by copper and iron dyshomeostasis. Currently, the copper and zinc chelating agent clioquinol represents a potential therapeutic route that may not only inhibit beta-amyloid neurotoxicity, but may also reverse the accumulation of neocortical beta-amyloid. A Phase II double-blind clinical trial of clioquinol with B12 supplementation will be published soon, and the results are promising. This article summarizes the role of transition metals in amyloidgenesis and reviews the potential promise of chelation therapy as a treatment for AD. PMID- 12890081 TI - Ethical issues in end-of-life geriatric care: the approach of three monotheistic religions-Judaism, Catholicism, and Islam. AB - Ethical dilemmas pervade modern geriatric medicine. What is considered right or wrong will differ depending on, among other things, the patient's religion. The three Abrahamic monotheistic religions, Judaism, Christianity (its Catholic variant), and Islam all have carefully considered positions on medical ethics. Although much is held in common, there are significant differences. The authors present three clinical cases, each of which presents ethical dilemmas typical of geriatric care, especially at the end of life. On the basis of these scenarios, the normative ethical position of each religion is compared and contrasted. It is hoped that this approach will offer the geriatrician a useful approach to treating patients in an increasingly multicultural society. PMID- 12890082 TI - Using the family covenant in planning end-of-life care: obligations and promises of patients, families, and physicians. AB - Physicians and families need to interact more meaningfully to clarify the values and preferences at stake in advance care planning. The current use of advance directives fails to respect patient autonomy. This paper proposes using the family covenant as a preventive ethics process designed to improve end-of-life planning by incorporating other family members--as agreed to by the patient and those family members--into the medical care dialogue. The family covenant formulates advance directives in conversation with family members and with the assistance of a physician, thereby making advance directives more acceptable to the family, and more intelligible to other physicians. It adds the moral force of a promise to the obligation of respecting a patient's preferences about end-of life care. These negotiations between patient, family, and physician, from early planning phases through implementation, should greatly reduce the incidence of family disagreements on what the patient would have wanted. The family covenant ensures advance directive discussions within the family, promotes and respects the autonomy of other family members, and might even spur others in the family to complete advance directives through additional covenants. The family covenant holds the potential to transform moral quagmires into meaningful moral conversation. PMID- 12890083 TI - Integrative review of physical activity intervention research with aging adults. AB - This paper reviews randomized, controlled trials (RCTs) that have attempted to increase physical activity behavior by aging adults. A systematic review was necessary because numerous studies target older adults, and previous reviews have addressed a limited range of primary studies. Computerized database, ancestry, and extensive search strategies by authors of research reported in English between 1960 and 2000 located diverse intervention trials. RCTs reporting endurance physical activity or exercise behavioral outcomes for at least five subjects were included. Integrative review methods were used to summarize extant research. Forty-two studies were retrieved. Seventeen RCTs with 6,391 subjects were reviewed. A wide variety of intervention strategies were reported. The most common interventions were self-monitoring, general health education, goal setting, supervised center-based exercise, problem solving, feedback, reinforcement, and relapse prevention education. Few studies individually adapted motivational interventions, used mediated intervention delivery, or integrated multiple theoretical frameworks into the intervention. Links between individual intervention components and effectiveness were not clear. Common methodological weaknesses included small samples, untested outcome measures, and time-limited longitudinal designs. Significant numbers of aging adults increased their physical activity in response to experimental interventions. The amount of increased activity rarely equaled accepted behavior standards to achieve positive health outcomes. Further work is essential to identify successful strategies to increase activity by larger numbers of elders and to accelerate the increase in activity by those who change activity behaviors. Sex and ethnic differences need further investigation. There is a vital need for rigorously designed studies to contribute to this science. PMID- 12890084 TI - Selected as the best paper in JAGS in the 1970s mission of the National Institute on Aging. AB - The National Institute on Aging (NIA), the newest of the 11 National Institutes of Health, is dedicated to improving the quality of life of the old in America through biomedical, social, and behavioral research. Aging is viewed as more than just decline and deterioration; it is also a process of continued development and accumulated knowledge. The NIA will encourage innovative research but will not support the delivery of health services, as that is the domain of the other agencies. In areas of overlap, such as diseases common to the old, the NIA will collaborate with other Institutes. A good target area for collaboration is senile dementia. Other areas of interest to the NIA are: encouraging the incorporation of geriatric medicine as a subspecialty, developing retirement test patterns, and investigating drug-drug and drug-age interactions, personality and social processes, and immunocompetence. PMID- 12890085 TI - The National Institute on Aging's mission is not 1970s nostalgia. PMID- 12890086 TI - Preventing falls in the elderly. PMID- 12890087 TI - Vascular disease, depression, and dementia. PMID- 12890088 TI - For the love of geriatrics. PMID- 12890089 TI - Severe acute respiratory syndrome (SARS) in elders. PMID- 12890090 TI - How do older people react to severe acute respiratory syndrome? PMID- 12890091 TI - Tetraplegia in a patient with huge heterotopic ossification of the hip joint after total hip arthroplasty: addition or reinforcement of handicaps. PMID- 12890092 TI - Clinical features of influenza a virus infection in older hospitalized persons. PMID- 12890093 TI - Quantification of taiji learning in older adults. PMID- 12890094 TI - Bedside physician objects. PMID- 12890096 TI - Age-related decline of gait control under a dual-task condition. PMID- 12890097 TI - Refilling prescriptions by community-dwelling elderly. PMID- 12890099 TI - Celiac disease and dermatitis herpetiformis: the spectrum of gluten-sensitive enteropathy. PMID- 12890100 TI - Dermatitis herpetiformis. PMID- 12890101 TI - Incidence of anti-Helicobacter pylori and anti-CagA antibodies in rosacea patients. AB - In recent years some authors have reported a possible correlation between Helicobacter pylori (Hp) and dermatological diseases such as rosacea. In this study we evaluated serum IgG and IgA anti-Hp in a group of 48 patients with rosacea. IgG antibodies were present in 81% of the rosacea patients with dyspepsia and 16% of the rosacea patients without dyspeptic symptoms. Serum IgG and IgA anti-Hp were detected by means of an enzymatic immunoabsorption test (enzyme-linked immunosorbent assay: ELISA). In addition, an evaluation of the anti-CagA antibodies by means of an immunoenzymatic method was carried out (ELISA, RADIM). IgA anti-Hp was present in 62% of patients with dyspepsia and in 6% of patients with no upper gastrointestinal symptoms. Anti-CagA antibodies were seen to be present in 75% of patients with both rosacea and gastric symptomatology, and were prevalent in patients affected by rosacea with papular symptoms in respect to rosacea with erythematous symptoms. The data that emerge from our case studies appear to confirm the hypothesis put forward by scientists who propose a correlation between the two clinical presentations rather than an inverse hypothesis of total independence. PMID- 12890102 TI - Common allergens in shoe dermatitis: our experience in Lahore, Pakistan. AB - BACKGROUND: Shoe dermatitis is a form of contact dermatitis resulting from exposure to shoes. Different chemicals, in conjunction with a hot and humid environment within the shoe, give rise to allergic or irritant dermatitis. Allergic shoe dermatitis is commonly caused by constituents of rubber, leather, adhesives, and rarely by linings and dyes. AIM: To determine the frequency of various allergens in shoe dermatitis in our patients. METHODS: One hundred and nine patients suspected of having contact dermatitis due to shoe allergens were included. All patients were patch tested with the Shoe series and European Standard series. Patches were applied on the upper back and removed after 48 h. Results were read at 48, 72, and 120 h and interpreted according to International Contact Dermatitis Research Group (ICDRG) criteria. RESULTS: Of the 119 patients, 87 (73%) reacted positively to various allergens, 48% of whom showed polysensitivity. Glues (33.6%), particularly para-tert-butylphenol formaldehyde resin (PTBP-FR) (26.9%), were the leading cause of shoe dermatitis. Other allergens were leather chemicals (26.4%), potassium dichromate (16.18%), rubber allergens (7.6%), and dyes (7.6%). Shoe scrapings tested positive in 26.3% of patients. Other common allergens were nickel sulfate (22.7%), neomycin sulfate (10.1%), and cobalt chloride (8.4%). CONCLUSIONS: Considering the diversity of allergens, all patients presenting with shoe dermatitis should be patch tested with the Shoe series and a standard series to determine the etiologic allergens. PMID- 12890103 TI - Paederus dermatitis in northern Iran: a report of 156 cases. AB - BACKGROUND: Paederus dermatitis develops when beetles of the genus Paederus (often called rove beetles) are crushed on the skin, releasing the vesicant pederin. These beetles are found in many tropical and subtropical habitats. METHODS: We describe 156 patients who presented to a dermatology clinic in the Guilan province of northern Iran during a 6-month period (May-October 2001). RESULTS: The peak time of presentation was in September, and the face and neck were the most common sites of involvement. Clinically, the most common presentation comprised geographic erythematous plaques with micropustules. In three-quarters of patients, more than one lesion was present. Kissing lesions were seen in 5% of cases, and 15% of patients developed diffuse desquamation. The majority of patients resided within 1 km of rice fields and used fluorescent lighting at home. In half of the cases, another family member was also affected. CONCLUSIONS: Paederus dermatitis is a common skin condition in northern Iran. We believe that increased public awareness of this condition can decrease mucocutaneous exposure to pederin. PMID- 12890104 TI - Comparison of chronic autoimmune urticaria with chronic idiopathic urticaria. AB - BACKGROUND: Chronic urticaria has been described in patients with Helicobacter pylori infection. Despite numerous studies, the correlation between H. pylori infection and chronic urticaria is doubtful. Our study was performed to determine the prevalence of H. pylori infection in autoimmune urticaria and in patients suffering from autoimmune urticaria and autoimmune thyroiditis. METHODS: The authors widely investigated 48 patients. The examinations were extended principally to autologous serum skin test, antithyroid antibodies, and the presence of H. pylori infection as well as detection of antibodies against H. pylori. RESULTS: Out of the 48 patients, 26 were regarded as having autoimmune origin. The prevalence of antithyroid antibodies was different in the two groups of patients with urticaria. There were 11 patients (42.3%) in the autoimmune group compared with three patients (13.6%) in the nonautoimmune group with antithyroid peroxidase antibody (P = 0.03). The difference in the prevalence of H. pylori infection was significant between autoimmune urticaria with and without thyroid autoimmunity (90.9% vs. 46.7%; P = 0.02). Autoimmune thyroiditis was connected with CagA +H. pylori strains, as the H. pylori- specific IgG antibodies revealed significant differences in a prevalence of 120 kDa (P < 0.05). CONCLUSIONS: The authors observed a relationship between autoimmune urticaria and autoimmune thyroiditis. The results strengthen the possibility of cross reactivity being triggered between CagA plus H. pylori strains and some other organ-specific autoimmune diseases such as autoimmune urticaria and autoimmune thyroiditis. This indicates a possible role of H. pylori in triggering autoimmune urticaria in at least a select group of patients. PMID- 12890105 TI - Complementary and alternative medicine use among patients attending a hospital dermatology clinic in Taiwan. AB - BACKGROUND: Recent studies have indicated that complementary and alternative medicine (CAM) has been gaining popularity around the world. Previous studies showed that CAM use was common among dermatology patients in Western nations. This study presents the prevalence and types of CAM use among a group of patients in a dermatology clinic in Taiwan. METHODS: A convenient sample of 198 patients was recruited from the dermatology clinic of Show Chwan Memorial Hospital in Changhua City, Taiwan. Each patient completed a standardized questionnaire. Chi square tests were used to compare the demographic characteristics of those who did and did not use CAM. Logistic regression analyses were performed to determine the strength of association between correlate variables and CAM use. RESULTS: Forty-one per cent of subjects had used CAM to treat their dermatological conditions. Subjects in the younger (< 30 years) and older (> 50 years) age groups were more likely to use CAM than those in the 30-50-year age group. Those that participated in religious or social groups were more likely to use CAM. CAM users felt a more negative impact of their dermatological conditions on daily living, relationships, emotional well-being and physical health. CONCLUSIONS: As in Western nations, CAM use is prevalent among dermatology patients in Taiwan: not only the traditionally popular Chinese CAM, but also Western CAM. With CAM being popular in the younger age group, dermatologists will likely encounter more CAM users in the future. Because some CAM may produce serious side-effects, dermatologists need to help and guide patients to assess the benefits and dangers of using CAM to treat dermatological conditions. PMID- 12890106 TI - Lymphoscintigraphic analysis in chromoblastomycosis. AB - BACKGROUND: One of the main complications of chromoblastomycosis is lymphedema. The purpose of this study was to evaluate the lymphatic system of the limbs of patients with chromoblastomycosis using lymphoscintigraphy. It is a reliable, objective and noninvasive means of supporting the diagnosis of lymphedema. METHODS: Lymphoscintigraphy was performed in seven patients with chromoblastomycosis, six with lesions in the lower limb and one in the upper limb. Tc-99 m dextran was injected into the interdigital spaces of the upper or lower extremities. The qualitative parameters analyzed were the visibilization of the lymph vessels and the lymph nodes, dermal backflow, and existence of collateral vessels. All patients were treated with the association of itraconazole and cryotherapy with liquid nitrogen. RESULTS: Three out of the 14 extremities examined had lymphedema clinically, and the lymphoscintigraphy showed abnormalities in the qualitative parameters; whereas these parameters were normal in the extremities without lymphedema. During the treatment of chromoblastomycosis, a second lymphoscintigraphy exam was performed on 10 limbs and did not show any improvement of the previous lymphoscintigraphic alterations. CONCLUSIONS: The qualitative lymphoscintigraphy was a reliable method to show the morphology of the lymph vessels and confirm objectively the diagnosis of lymphedema secondary to chromoblastomycosis. The association of oral itraconazole and cryotherapy did not modify the lymphatic alterations in chromoblastomycosis. PMID- 12890107 TI - Head lice on pillows, and strategies to make a small risk even less. AB - BACKGROUND: Due to a lack of evidence, controversy exists about the role of bedding in the transmission of head lice. AIMS: To determine the proportion of the head lice population found on pillowcases of people with head lice, and to test strategies available to householders to kill head lice on pillowcases. METHODS: To assess the incidence of head lice on pillowcases, people with active pediculosis had their head lice collected and counted and the pillowcase they had used the night before examined for head lice. To test strategies to kill head lice on pillowcases, live head lice were experimentally placed in miniature pillowcases, and the cases were subjected to a hot wash, a cold wash, hot dryer, and hanging out to dry on an outdoor clothes line. RESULTS: Forty-eight people and their pillowcases were recruited from Townsville, Qld, Australia (dry tropics). One thousand, eight hundred and forty-five lice were collected from their heads to give an average and median intensity of infection of 38.4 and 21 lice, respectively. Two of the 48 pillowcases contained live lice, one nymph on each, 2 h and 9 h after the pillowcases had been removed from the bed. Another pillowcase contained a dehydrated nymph. The incidence of live lice on pillowcases was 4.2% per night and the proportion of the head louse population on the pillowcases was 0.11%. Heat (hot wash and hot clothes dryer) killed head lice experimentally placed in pillowcases. Cold wash and hanging pillowcases out to dry did not kill head lice. CONCLUSIONS: Head lice transfer to pillowcases at night, but the incidence is low. Pillowcases pose a risk for re-infection with head lice, but the risk is low, and changing the pillowcase is a reasonably cost efficient strategy to minimize this risk. Lice on pillowcases can be killed by heating the pillowcase by immersion in water at > 60 degrees C, by a hot wash, or by 15 min in a hot clothes dryer. PMID- 12890108 TI - Cutaneous alternariosis in a renal transplant recipient. PMID- 12890109 TI - Is topical metronidazole effective in seborrheic dermatitis? A double-blind study. AB - BACKGROUND: Seborrheic dermatitis is a common, chronic, papulosquamous dermatosis. Although several types of treatment have been developed, repetitive treatment courses are often necessary. AIM: To perform a randomized, placebo controlled, double-blind clinical study with metronidazole 0.75% gel in patients with mild to moderate seborrheic dermatitis of the face. METHODS: Eighty-four patients (52 males and 32 females) with seborrheic dermatitis were enrolled in the study. Patients were randomly allocated to the application of metronidazole 0.75% gel or placebo to their facial lesions, twice daily for 8 weeks. Patients were scored with regard to the severity of the lesions at the initial evaluation and every 2 weeks for 2 months. All patients were evaluated for a final assessment of improvement at the end of the study. RESULTS: At baseline, both treatment groups were comparable in terms of demographic data and lesion severity score. Seventy-eight patients (48 males and 30 females), 48 (62%) in the metronidazole group and 30 (38%) in the placebo group, completed the study. Two patients in the metronidazole group and four patients in the placebo group left the study. There was no statistically significant difference in the change in the mean severity score between the two groups at the end of the study (P > 0.05). At the final evaluation of the response at 8 weeks, no statistically significant difference was found between the treatment groups (P > 0.05). Metronidazole gel was generally tolerated well during the study. CONCLUSIONS: Metronidazole 0.75% gel and placebo show similar efficacy in the treatment of seborrheic dermatitis. PMID- 12890110 TI - Histoplasmosis presenting as erythroderma in a patient with the acquired immunodeficiency syndrome. PMID- 12890111 TI - Nodular vasculitis as a paraneoplastic presentation? PMID- 12890112 TI - Mycobacterium chelonae infection with multiple cutaneous lesions after treatment with acupuncture. PMID- 12890113 TI - How safe is nose-piercing? Inoculation cutaneous tuberculosis revisited. PMID- 12890114 TI - Kaposi's sarcoma after allogeneic peripheral blood stem cell transplantation. PMID- 12890115 TI - Case of generalized lichen amyloidosis. PMID- 12890116 TI - Waardenburg syndrome: case report. PMID- 12890117 TI - Eccrine porocarcinoma arising in a seborrheic keratosis evaluated with dermoscopy and treated with Mohs' technique. AB - A 78-year-old white woman returned for a routine 6-month skin cancer examination. She had a history of actinic keratosis and multiple basal cell carcinomas. She had no personal or family history of dysplastic nevi or melanoma. The patient was asymptomatic and unaware of any new or changing skin lesions. The patient had multiple lentigines, hemangiomas, and actinic and seborrheic keratoses on all sun exposed areas. There were no less than 10 seborrheic keratoses on the right mid back, and one was found to have a 1-cm, reddish nodule asymmetrically located within it (Figs 1 and 2). A clear papule on the left preauricular area was found on biopsy to be a basal cell carcinoma. The nodule on the back was still present 1 month later and it was felt that further evaluation was indicated. As melanoma has been reported to develop in seborrheic keratoses, we decided to examine the lesion using digital dermoscopy. With digital dermoscopy, a well-demarcated reddish nodule was asymmetrically located within a brown lesion. It blanched significantly with pressure. Within the nodule, there were dotted and irregular linear vessels (atypical vascular pattern; also known as polymorphous vascular pattern) and regular-appearing brown dots. Surrounding the reddish nodule, there were pale and pigmented, comedo-like openings, fissures, and ridges (brain-like appearance). Some of the follicular openings appeared to be within the wall of the nodule (Figs 3 and 4). Comedo-like openings, fissures, and ridges are primary dermoscopic criteria for the diagnosis of a seborrheic keratosis; however, the vascular pattern seen has not been reported in seborrheic keratosis. Due to the patient's age and the rarity of significant pathology arising in a seborrheic keratosis, a shave biopsy was performed. To our surprise, the specimen was interpreted by an experienced dermatopathologist as a well-differentiated eccrine porocarcinoma. Due to the high local recurrence rate and metastatic potential of this carcinoma, the patient was referred for Mohs' surgery. Both the basal cell carcinoma and the eccrine porocarcinoma were excised in one stage. A metastatic work-up was negative and the patient appears to be doing well. PMID- 12890118 TI - 308-nm excimer laser for the treatment of localized vitiligo. AB - BACKGROUND AND OBJECTIVE: Vitiligo is commonly treated with PUVA, and more recently, narrow-band UVB (NBUVB) phototherapy. Given the proximity of the wavelengths of NBUVB (311 nm) and the excimer laser (308 nm), we undertook a clinical trial to test the efficacy of this device. METHODS: Twice-weekly 308-nm UV-B radiation was given to selected vitiligo lesions for a maximum of 60 treatments. These lesions had been unsuccessfully treated previously with at least one other method of treatment. Initial doses were 100 mJ/cm2 with increments of 10-25%. Improvement was assessed on a visual scale via serial photographs. RESULTS: Subjects tolerated the treatment well. Improvement varied with body site. After 60 treatments, lesions on the hands and feet showed grade 2 improvement in 2/10 subjects and grade 1 in 8/10. For the axillae, there was grade 4 improvement in 1/3 subjects and grade 2 improvement in 2/3 by treatment 60. The face demonstrated the most rapid repigmentation with grade 4 repigmentation seen in 3/5 subjects by 40 treatments and grade 3 in 2/5 by 30 treatments. There were no adverse effects. CONCLUSIONS: The user-friendly 308-nm excimer laser allows targeted treatments of localized vitiligo. PMID- 12890121 TI - Wolf's isotopic response: herpes simplex appearing on scrofuloderma scar. PMID- 12890122 TI - Household cluster of tinea capitis caused by Trichophyton violaceum in Brazil. PMID- 12890124 TI - 2003 Wolff Award: Possible parasympathetic contributions to peripheral and central sensitization during migraine. AB - BACKGROUND: Neurologic signs of increased parasympathetic outflow to the head often accompany migraine attacks. Because increased parasympathetic outflow to the cranial cavity induces vasodilation of cerebral and meningeal blood vessels, it can enhance plasma protein extravasation and the release of proinflammatory mediators that activate perivascular nociceptors. We recently showed that activation of intracranial perivascular nociceptors induces peripheral and central sensitization along the trigeminovascular pathway and proposed that these sensitizations mediate the intracranial hypersensitivity and the cutaneous allodynia of migraine. METHODS: The present study investigates possible parasympathetic contributions to the generation of peripheral and central sensitization during migraine by applying intranasal lidocaine to reduce cranial parasympathetic outflow through the sphenopalatine ganglion. RESULTS: In the absence of migraine, patients were pain-free, and their skin sensitivity was normal. Their mean baseline pain thresholds were less than 15 degrees C for cold, more than 45 degrees C for heat, and more than 100 g for mechanical pressure. Their mean pain score was 7.5 of 10 (standard deviation, 1.4) during untreated migraine and 3.5 of 10 (standard deviation, 2.4) after the nasal lidocaine induced sphenopalatine ganglion block (P <.0001). Most patients developed cutaneous allodynia during migraine, and their mean pain thresholds changed to more than 25 degrees C for cold, less than 40 degrees C for heat, and less than 10 g for mechanical pressure. Following the nasal lidocaine administration (sphenopalatine ganglion block), this allodynia remained unchanged in spite of the pain relief. CONCLUSION: These findings suggest that cranial parasympathetic outflow contributes to migraine pain by activating or sensitizing (or both) intracranial nociceptors, and that these events induce parasympathetically independent allodynia by sensitizing the central nociceptive neurons in the spinal trigeminal nucleus. PMID- 12890125 TI - Headache management program improves outcome for chronic headache. AB - OBJECTIVE: To determine the feasibility of developing a headache management program and to assess the outcomes of patients referred to the program for treatment of chronic headache. BACKGROUND: Effective headache treatment requires that the patient receives the correct headache diagnosis; that appropriate acute and, if indicated, preventive medications be prescribed; and that the patient receives adequate education, including headache self-management skills. DESIGN/METHODS: A headache management program was established at a northern California staff-model health maintenance organization. Fifty-four patients were enrolled in the program and followed for 6 months. Patients participated in a structured program of group and individual sessions with the program manager. Data collection at baseline and 6 months included the Migraine Disability Assessment (MIDAS), the Short Form-36 Health Survey (SF-36), a patient satisfaction survey, and 2 additional short surveys--one that assessed patient worries about their headaches and another that queried patients on their problems with headache management. RESULTS: All enrolled patients participated in the initial group visit; 74% had at least one additional visit. All but one patient suffered from more than one headache type. Sixty-one percent of patients suffered from migraine headache and 98% from tension-type headache. At baseline, patients were severely disabled, with a mean MIDAS score of 41. At 6 months, MIDAS scores decreased an average of 21.2 points (P <.005). Patients reported 14.5 fewer days with headache over the preceding 3 months (P <.0001) and experienced clinically significant improvements in 6 of the SF-36 subscales. Patients were significantly more satisfied with their headache care (P <.0001), reported less problems with their headache management (P <.0001), and were less worried about their headaches (P <.01). During the intervention, emergency department visits for headache decreased (P <.02). CONCLUSIONS: A headache management program was successfully established. Patients referred to the program experienced significant improvement in headache-related disability and functional health status and reported greater satisfaction with care. Even so, these results were obtained at one site and in a small sample that was not randomized. We currently are conducting a randomized controlled trial to better evaluate the clinical and financial impact of a headache management program for patients with chronic headache. PMID- 12890126 TI - Prevalence of atrial septal aneurysm in patients with migraine: an echocardiographic study. AB - OBJECTIVE: To evaluate the prevalence of atrial septal aneurysm in patients with migraine. BACKGROUND: Migraine has long been considered a risk factor for stroke. Atrial septal aneurysm is often observed in young patients with ischemic stroke and is frequently associated with other conditions potentially leading to embolism. METHODS: We performed a transthoracic echocardiogram in 90 consecutive patients (65 women and 25 men; mean age, 35.3 years [standard deviation, 9]) with migraine but free from cerebral and cardiovascular disease and in 53 control subjects (37 women and 16 men; mean age, 34 years [standard deviation, 10]). The diagnosis of atrial septal aneurysm was performed according to Olivares-Reyes criteria. A transesophageal echocardiogram also was performed in 75 patients with migraine (83.3%). RESULTS: The prevalence of isolated atrial septal aneurysm was higher in patients with migraine with aura (28.5%) than in patients with migraine without aura (3.6%) (P <.005) or in control subjects (1.9%) (P <.005). CONCLUSIONS: Our data suggest a role of atrial septal aneurysm in the genesis of aura in patients with migraine. PMID- 12890127 TI - Sumatriptan plus metoclopramide in triptan-nonresponsive migraineurs. AB - OBJECTIVES: We evaluated the effectiveness of combination treatment using sumatriptan plus metoclopramide versus sumatriptan alone for the treatment of acute migraine. The patients who were treated had failed to respond to triptans in the past despite adequate doses on at least 2 separate trials of the same triptan or 2 trials involving different triptans. BACKGROUND: There is limited evidence that dopaminergic antagonists may benefit the migraineur by relieving migraine pain and associated symptoms. The exact mechanism of action in migraine is unknown. The postulated action is the inhibition of dopaminergic overactivity. A dopaminergic antagonist, metoclopramide, may improve the efficacy of a 5 HT1B/1D agonist, sumatriptan. METHODS: In this double-blind, randomized, crossover study, 16 adult migraineurs fulfilling International Headache Society (IHS) criteria for migraine with or without aura who had failed to receive adequate relief from triptans treated one migraine with each treatment: sumatriptan 50 mg plus metoclopramide 10 mg or sumatriptan 50 mg plus placebo to match metoclopramide. Patients treated their migraines when they were moderate or severe in intensity and recorded pain severity and symptoms prior to treatment and 30, 60, 90, and 120 minutes and 24 hours after treatment. RESULTS: Thirteen women and 3 men (mean age, 40 years) completed the study; ie, treated 2 migraines (a total of 32 migraines), one attack with each treatment. Meaningful relief was attained in 10 (63%) of 16 migraines treated with the combination of sumatriptan 50 mg plus metoclopramide 10 mg compared with 5 (31%) of 16 migraines treated with sumatriptan 50 mg plus placebo. Headache response (moderate or severe to mild or no pain at 2 hours) was achieved in 7 (44%) of 16 migraines with the combination of sumatriptan 50 mg plus metoclopramide 10 mg compared with 5 (31%) of 16 migraines treated with sumatriptan 50 mg plus placebo. There did not appear to be a difference between treatment groups with respect to associated symptoms. The combination of sumatriptan 50 mg plus metoclopramide 10 mg was well tolerated. CONCLUSIONS: Combining sumatriptan with metoclopramide provided relief in some migraineurs who failed to achieve adequate relief with a triptan alone. It remains unknown whether initiating therapy when pain was mild or using a higher dose of sumatriptan (ie, 100 mg) would have provided additional benefit. Further studies are indicated. PMID- 12890128 TI - Almotriptan versus rizatriptan in patients with migraine in Spain. AB - OBJECTIVES: To compare patient-reported use of rizatriptan 10 mg with that of almotriptan 12.5 mg per migraine attack (24 hours) in a Spanish population. METHODS: One hundred twenty Spanish community pharmacies recruited patients with migraine to whom they had dispensed almotriptan and rizatriptan. No other selection criteria were used. Patients kept diaries for baseline pain intensity, the number of triptan tablets used, additional medication taken per attack, and their degree of satisfaction with the medication 2 hours after the initial dose. Patients recorded details for a maximum of 3 attacks. Analysis of variance or the Student t test and chi-squared or Fisher exact tests were used for univariate comparisons. A generalized estimating equation method was used to correct for within-subject variability. Adjusted odds ratios (ORs) and 95% confidence intervals (CIs) were calculated. RESULTS: One hundred twenty-six patients (85% women) recorded data for 318 migraine attacks. Rizatriptan was used to treat 122 attacks, almotriptan was used to treat 110 attacks, and a nontriptan medication was used in the initial treatment of 86 attacks. Triptan use (adjusted mean, 95% CI) per attack in this study was lower for rizatriptan (1.19 tablets; 95% CI, 1.06 to 1.32) than for almotriptan (1.43 tablets; 95% CI, 1.30 to 1.56; P=.003). The use of a triptan and additional medication per attack increased with baseline pain severity. Rizatriptan was used to treat more attacks with only one tablet (78%) than almotriptan (58%). Treatment of attacks with almotriptan was more than twice as likely to involve the use of more than one tablet per attack (24 hours) than those treated with rizatriptan (adjusted OR, 2.42; 95% CI, 1.37 to 4.30; P=.003). Patient satisfaction with treatment response at 2 hours was more than 2 fold greater for rizatriptan (85%) than for almotriptan (68%) (adjusted OR, 2.55; 95% CI, 1.11 to 5.87; P=.03). CONCLUSIONS: In this prescription-selected Spanish population, a significantly lower number of rizatriptan tablets were required to treat migraine attacks compared with almotriptan. Further, patients were more than twice as likely to use more than one tablet or additional medication (or both) for attacks treated with almotriptan than for those treated with rizatriptan. Although these data suggest that rizatriptan may be a more effective treatment for migraine than almotriptan, further randomized studies are required to confirm this conclusion. PMID- 12890129 TI - Burden of migraine in Brazil: estimate of cost of migraine to the public health system and an analytical study of the cost-effectiveness of a stratified model of care. AB - BACKGROUND: The search for rationality in health expenses in developing countries collides with the lack of effectively conducted epidemiologic studies. PURPOSE: To present an estimate of the impact and costs of migraine in the Brazilian public health system and to estimate the impact on these costs and the effectiveness of a model of stratified care in the management of migraine. METHODS: An analytical model of utilization of the Brazilian public health system was constructed. Data refer to 1999 and were obtained in accordance with the following steps: (1) Brazilian demographic characteristics; (2) characteristics of the public health system related to its 3 hierarchical levels-primary, secondary, and tertiary care, the last being subdivided into emergency department and hospital care; and (3) estimation of the number of migraine consultations at each complexity level. In Brazil, migraineurs seen in the public health system are most often discharged with an acute treatment, usually a nonspecific medication. We compared this treatment with a proposed stratified care model that uses a triptan as an acute care medication. We have made the following assumptions: (1) 15% of the patients would fall into the Migraine Disability Assessment (MIDAS) grade I category, 25% would fall into the MIDAS grade II category, 30% into the grade III category, and 30% into the MIDAS grade IV category; (2) the mean number of migraine attacks per year are: MIDAS I, 7.49; MIDAS II, 8.02; MIDAS III, 12.22; and MIDAS IV, 27.01. The annual costs of the treatment were calculated according to the following equation: AC = P x N x C + P x Cp + P x Cat x AMA, where P is the number of patients; N, the number of consultations per patient; C, the cost of consultation per level; Cp, the cost of preventive drugs; Cat, the cost of acute therapy drugs; and AMA is the number of migraine attacks per year. Results.-The public health system resources included 55 735 ambulatory units (primary and secondary) and 6453 emergency department and public hospital units, with a corresponding budget of US $2 820 899 621.26. The estimated cost of a consultation on the primary care level was US $11.53; on the secondary care level, US $22.18; in the emergency department, $34.82; and for hospitalization, US $217.93. The total estimated public health system expenses for migraine were US $140 388 469.60. The proposed model would imply a cost reduction of 6.2% (US $7 514 604.40) with an improvement in the quality of the public health system from the actual 18.2% to an estimated 84.5%. CONCLUSION: Migraine seems to pose a huge burden on the Brazilian public health system. The implementation of a stratified care model of treatment that would include specific acute migraine therapies could result in a dramatic increase in the quality of migraine care and a significant reduction in cost. PMID- 12890130 TI - Posttraumatic headache: biopsychosocial comparisons with multiple control groups. AB - OBJECTIVE: This study examined somatic, psychological, and cognitive functioning of subjects with posttraumatic headache in comparison with multiple control groups. BACKGROUND: Posttraumatic headache is not as widely studied as other forms of headache (eg, tension-type, migraine). Previous research has suggested poor psychological functioning in patients with posttraumatic headache in comparison with other groups of patients with pain; however, this group has yet to be compared with a group of persons who have experienced trauma but are headache-free. DESIGN AND METHODS: Nineteen subjects with posttraumatic headache were studied, with full assessments available for 14 participants. Comparison groups, containing 16 participants each, included another headache group, a nonheadache group, and a trauma (motor vehicle accident) survivor nonheadache group. Participants completed several measures assessing somatic, psychological, and cognitive functioning. RESULTS: Findings revealed that the posttraumatic headache group exhibited significantly poorer functioning than the comparison groups on several measures including the Psychosomatic Symptom Checklist, Postconcussion Syndrome Checklist, axis II psychiatric diagnoses, Minnesota Multiphasic Personality Inventory, and the Daily Hassles Scale (frequency and total). Additionally, they scored higher on the following: number of axis I psychiatric diagnoses, the Daily Hassles Scale (intensity), Beck Depression Inventory, State-Trait Anxiety Inventory, and State-Trait Anger Expression Inventory. The posttraumatic headache group was similar to the other trauma group on the Posttraumatic Stress Disorder Symptom Checklist and the Life-Trauma Checklist. CONCLUSIONS: This study confirmed the distress seen in this understudied population of persons with headache and highlights areas of focus for proper assessment and treatment of those with headache and who have had an accident. PMID- 12890131 TI - Use of the Migraine Disability Assessment Questionnaire in children and adolescents with headache: an Italian pilot study. AB - OBJECTIVES: To determine the suitability of the Migraine Disability Assessment (MIDAS) Questionnaire for assessing disability in children and adolescents with headache and to obtain preliminary information about disability in different primary headaches. BACKGROUND: During the last decade, researchers have begun to employ standardized methodologies to investigate the global impact of primary headaches. Disease-specific instruments have been developed to measure headache related disability. The MIDAS Questionnaire, which is the most extensively studied of these instruments, was designed to assess the overall impact of headaches over the 3 months before compilation. The MIDAS Questionnaire is an optimal tool to assess headache-related disability in adults. METHODS: Ninety five patients aged 7 to 17 years with tension-type headache, migraine, or both completed the validated Italian form of the MIDAS questionnaire on 2 occasions. Test-retest reliability was assessed by the Spearman rank correlation test. The Cronbach alpha assessed internal consistency. The patients answered questions about the adequacy of the questionnaire. RESULTS: The Cronbach alpha was.8. Correlation coefficients were generally high for the overall MIDAS score and for the items investigating disability in school and in family/leisure activities; they were lower for the items about housework. Most patients thought that the MIDAS Questionnaire was useful (98.9%) and that it captured the impact of their headaches (58.9%); 41% thought that questions about disability in housework were useless, 44.2% suggested adding questions regarding inability to do homework. All primary headaches had a considerable impact on daily activities, but patients with migraine tended to have lower headache frequencies and lower total disability time; those with tension-type headache suffered more days in which activities, although performed, were substantially impaired. CONCLUSIONS: The MIDAS Questionnaire is useful for assessing disability in children and adolescents with different primary headaches. Minimal changes in the phrasing and content of the items would be sufficient to render the MIDAS specific for the younger population with headache. PMID- 12890132 TI - Prevalence of headache in Puerto Rico. AB - BACKGROUND: Headache is one of the most frequently reported disorders in the general adult population. Despite the fact that this disorder is common, no official estimate of the prevalence of headache exists in Puerto Rico. PURPOSE: To examine the prevalence of headache and migraine, specifically, in Puerto Rico. METHODS: A telephone survey of 1610 individuals was conducted. The telephone calls were distributed using the 1990 census adjusted to the population of Puerto Rico in 1998, and according to gender, geographical area, and age. RESULTS: The prevalence of headache in Puerto Rico was 35.9% and migraine, in particular, 13.0%. When prevalence was fractionated by age, gender, and geographical areas, the prevalence of headache was similar for all ages, with females exhibiting a 2:1 preponderance over males. In the younger population (between 20 and 50 years of age), the female-male ratio for migraine was 3:1, and the prevalence for migraine was lower in the Metropolitan area. CONCLUSION: This study, the first of its type in Puerto Rico, demonstrates that headache (and migraine specifically) is a common disorder in this country. PMID- 12890133 TI - Aspects of referral care for headache associated with improvement. AB - OBJECTIVE: To assess which aspects of referral care for headache are associated with improvement of pain and subjective quality of life. BACKGROUND: In managed care, referrals to a specialist are sometimes kept to a minimum. It has been questioned whether patients with headache do better after consultation with a specialist. METHODS: We mailed a questionnaire to all patients referred for headache to a neurologic center in northern Norway during a 2-year period (n = 1403). The questionnaire included items concerning diagnosis and treatment, along with simple visual analog scales to assess whether the patient's headache syndrome and self-perceived quality of life had changed after seeing the specialist. RESULTS: There were 1052 responders (75%). Headache generally decreased after consultation with a specialist; it decreased significantly more in the 527 patients who were assigned a diagnosis compared to the 344 patients who claimed they were not. Reduction of headache also was significantly more obvious in the 483 patients who had treatment prescribed, as compared to the 385 patients not receiving any therapeutic measure. Self-perceived quality of life was generally improved, significantly more when the patient was given a diagnosis, and even when the diagnosis did not lead to treatment. CONCLUSIONS: Patients referred to a neurologic center for evaluation of headache generally experience a significantly greater improvement in their headache syndrome and quality of life. This appears particularly so when they receive a diagnosis, even if no treatment is prescribed. PMID- 12890134 TI - Topiramate in the prophylactic treatment of cluster headache. AB - OBJECTIVE: The role of topiramate in the prophylactic treatment of cluster headache is still unclear. The aim of this study was to evaluate the effectiveness of topiramate in a group of patients with refractory episodic or chronic cluster headache. BACKGROUND: Proof of efficacy of preventive treatment of cluster headache is limited, especially for the chronic form of the disorder. There are very few randomized clinical trials on this condition with topiramate or other new anticonvulsant agents. Recent case reports and series involving topiramate have shown good results. The mechanism of action of topiramate is unknown, but is presumably mediated by gamma-aminobutyric acid. METHODS: Twenty six patients with episodic (n = 12) or chronic (n = 14) cluster headache were studied prospectively. All patients had been treated with some preventive treatment, with poor or no response. Treatment with topiramate was initiated with 25 mg once a day, and the dose was titrated every 3 to 7 days to a maximum of 200 mg, according to clinical response and tolerability. RESULTS: Topiramate rapidly induced cluster remission in 15 patients, reduced the number of attacks more than 50% in 6 patients, and reduced the cluster period duration in 12. The mean time to remission was 14 days (range, 1 to 27), but in 7 patients remission was obtained within the first days of treatment with very low doses (25 to 75 mg a day). Tolerability was good within the lower range of doses used. Six patients discontinued treatment due to side effects (all with daily doses over 100 mg) or lack of efficacy. CONCLUSIONS: Our results confirm that topiramate can be an effective option for the preventive treatment of episodic and chronic cluster headache. PMID- 12890135 TI - Software reuse reference model approach in developing an automated medical information system (AMIS) for improving health care practice. AB - OBJECTIVE: To evaluate patient acceptance of an automated medical information system specific to headache. BACKGROUND: Studies suggest that automated tools may help health care delivery systems to be efficient and effective, but patient satisfaction remains a major concern. METHODS: We adapted our software reuse reference model and Unified Modeling Language to apply the domain model to the headache population. Patients with headache were tested both to validate the system and to evaluate patient satisfaction and headache management with an automated system. RESULTS: The mean age of all study participants was 44 years. Over 95% of the participants were satisfied or strongly satisfied with the Automated Medical Information System. CONCLUSIONS: The results strongly suggest that patients are willing and able to use nontraditional sources, such as the Automated Medical Information System, to learn about their illnesses. PMID- 12890136 TI - Acromegaly: a unique human headache model. PMID- 12890137 TI - Onset of ophthalmoplegic migraine with abducens palsy at middle age? PMID- 12890138 TI - Laughing headache: a novel type of triggered headache with response to divalproex sodium. PMID- 12890139 TI - Occipital (Arnold) neuralgia secondary to greater occipital nerve schwannoma. PMID- 12890140 TI - Not-so-benign sexual headache. PMID- 12890141 TI - QT prolongation with migraine medications. PMID- 12890142 TI - Droperidol and other neuroleptics/antiemetics for the management of migraine. PMID- 12890143 TI - Therapeutic gain or therapeutic ratio? PMID- 12890145 TI - Primary treatment with autologous stem cell transplantation in mantle cell lymphoma: outcome related to remission pretransplant. AB - OBJECTIVES: The aim of the first Nordic mantle cell lymphoma (MCL) protocol was to study the clinical significance of an augmented CHOP induction chemotherapy followed by high-dose chemotherapy and autologous stem cell transplantation (ASCT) and to examine the prognostic significance of stem cell contamination rates in newly diagnosed patients with MCL. PATIENTS AND METHODS: Forty-one newly diagnosed patients below 66 yr were enrolled and given three series of an augmented CHOP regimen. Responders underwent stem cell mobilization with a fourth course of CHOP, stem cell harvest and ASCT. Stem cell purging was optional in the protocol and followed the routine of each participating centre. The number of tumour cells in the reinfused autografts was estimated by flow cytometry or quantitative PCR. RESULTS: Induction therapy led to complete remission (CR) in 11 of 41 patients (27%), partial remission (PR) in 20 of 41 patients (49%) and no response in nine patients (22%), whereas one patient was not evaluable. Twenty seven of the 31 responders underwent ASCT and 24 achieved or maintained a CR. The overall and failure-free 4-yr survival on intention-to-treat basis were 51% and 15%, respectively. Among the transplanted patients, a significantly increased failure-free (P<0.03) and overall survival (P=0.03) was noted among patients transplanted in CR compared with PR, respectively. By contrast, reinfusion of highly variable numbers of tumour cells with the autografts (range 0.71-80 x 10(6) tumour cells), did not affect outcome. CONCLUSION: In MCL, an important strategy to improve the outcome will be to intensify the induction chemotherapy. PMID- 12890146 TI - DNA sequence profile of TP53 gene mutations in childhood B-cell non-Hodgkin's lymphomas: prognostic implications. AB - OBJECTIVES: The TP53 gene encodes a nuclear protein implicated in the regulation of the cell cycle, DNA repair, and apoptosis. TP53 mutations and other alterations have been described in numerous types of tumors, and some of these have been associated with poor prognosis. The aim of this study was to characterize TP53 mutations in childhood B non-Hodgkin's lymphoma, their correlation with clinical prognostic factors and response to therapy. PATIENTS AND METHODS: Samples from 49 children with B non-Hodgkin's lymphoma were examined for TP53 alterations by single-strand conformation polymorphism analysis (SSCP) of exons 5-9, direct sequencing and by p53 immunohistochemistry. RESULTS: Mutations of TP53 were detected in 11 of 49 (22.5%) patients and more specifically in 20% of Burkitt's lymphoma. The sequence analysis showed missense mutations in 10 cases and an insertion mutation in one case. Mutations of the transition type occurred more frequently than transversions (seven of 11). Analysis of the spectrum of single-nucleotide substitutions showed a 30% frequency of transition mutations in CpG dinucleotide sequences. The overall frequency of p53 immunostaining positivity was 36% (15 of 41). There was a very good agreement between protein expression and the presence of TP53 mutation (P=0.0005). No significant correlation was found regarding age, gender, clinical stage and LDH level and TP53 mutations. Comparison of EFS curves using the log rank test were also not significant. However, the analysis of the effects of mutations on the core p53 structure identified biological and biochemical mutants with phenotypes probably related to different response to chemotherapy. CONCLUSIONS: Our data suggest that some types of mutants can alter the protein distinctly and may be associated with a more aggressive phenotype. In addition, the impact of TP53 mutations on response to therapy may also be influenced by disruption of other genes. PMID- 12890147 TI - Gelatinases (MMP-2 and MMP-9), TIMP-1 expression and the extent of neovascularization in aggressive non-Hodgkin's lymphomas. AB - OBJECTIVES: The present study was carried out to clarify the role of matrix metalloproteinase-2 and -9 (MMP-2 and MMP-9), tissue inhibitor of metalloproteinase-1 (TIMP-1) and the extent of neovascularization in the clinicopathologic behavior of non-Hodgkin's lymphomas. METHODS: Paraffin-embedded histologic sections from 57 patients with aggressive non-Hodgkin's lymphomas were stained with MMP-2, MMP-9, TIMP-1, and factor VIII antibodies to correlate the expression of these markers to the clinical disease characteristics. RESULTS: Strong MMP-9 staining was found to be an adverse prognostic factor among patients with aggressive B-cell lymphomas, the probabilities for 5-yr disease-free survival being 73%, 63%, 50%, and 0% in patients with grades 0, 1, 2, and 3 staining, respectively. Among the patients with strong (grades 2 and 3) MMP-9 staining, however, positivity for TIMP-1 indicated a trend toward a more favorable prognosis. TIMP-1 expression also correlated with the immunoblastic and anaplastic lymphoma subtypes. The expression of the proteins for MMP-2 and factor VIII had no independent prognostic role. None of the study parameters correlated with disease stage, the occurrence of extranodal infiltrates, the occurrence of bulky tumor, or the IPI scores. CONCLUSIONS: Positivity for MMP-9 immunoreactive protein is an independent sign of an unfavorable prognosis in non-Hodgkin's lymphomas. This is not mediated through influences in tumor dissemination or neovascularization indicating it to carry other important biological functions. PMID- 12890148 TI - Expression levels of hsc70 and hsp60 are developmentally regulated during B-cell maturation and not associated to childhood c-ALL at presentation or relapse. AB - Heat shock proteins are potent regulators of apoptosis, and so they may also be involved in normal cellular differentiation and cancerogenesis. We used quantitative two-dimensional gel electrophoresis for determining whether either the constitutive chaperonic heat shock cognate protein 70 (hsc70) or heat shock protein 60 (hsp60) contribute to B-cell differentiation and leukemogenesis. We compared the expression of these hsps in normal peripheral blood (PB) CD19+ B cells, in pediatric bone marrow (BM) CD19+ CD10+ B-cell precursors (BCPs) from normal donors, and in BCPs from common acute lymphoblastic leukemia (c-ALL) patients at diagnosis and at relapse. We found that the mean levels of hsc70 in c ALL BCPs at initial presentation and at relapse failed to differ significantly. Likewise, they failed to differ significantly from the level in high-expressing normal BM BCPs or from that in low-expressing PB B-cells. Mean levels of hsp60 expression in c-ALL BCPs at initial presentation and at relapse were similar and not distinguishable from that in normal BM BCPs, however, elevated (by a factor of 2-3) compared with that in PB B-cells. Hsc70 and Hsp60 expressions were increased (by a factor of 2 of mean levels) in populations of normal BM BCPs as compared with populations of PB B-cells. Thus, no abnormal levels of hsc70 and hsp60 were detectable in populations of pediatric c-ALL BCPs neither at diagnosis nor at relapse. In contrast, our data were in support of developmentally regulated levels of hsc70 and hsp60 expression during B-cell ontogenesis. PMID- 12890149 TI - Danaparoid for heparin-induced thrombocytopenia: an analysis of treatment failures. AB - BACKGROUND: Patients with heparin-induced thrombocytopenia (HIT) (with or without thrombosis) require alternative anticoagulation because of their extreme risk of new thromboembolic complications. The first effective agent for this purpose may be danaparoid, a less-sulfated low molecular weight heparinoid. Recently, direct thrombin inhibitors have been used. OBJECTIVE: Five HIT patients, who developed new thromboembolic complications while receiving danaparoid, were analyzed to consider possible reasons for treatment failure and to promulgate strategies that improve efficacy. RESULTS: Three patients had acute HIT, one had recent HIT, and one with remote HIT was re-exposed to heparin during heart surgery. Danaparoid was started as intravenous bolus and infusion in one patient, and as 1250 units subcutaneously twice daily in four patients. The new complications that emerged on danaparoid were new venous thrombi in three patients (one with pulmonary emboli), lower extremity arterial thrombosis in one, myocardial ischemia in one, thromboembolic cardiovascular accidents in one, and fatal bowel necrosis in one (two patients suffered more than one complication). Platelet counts did not improve or worsened in four, improved partially in the other, and parameters of disseminated intravascular coagulation failed to improve in one patient. Four patients responded relatively dramatically when direct thrombin inhibitors were substituted. Possible reasons for danaparoid failure include that: 1) no treatment is expected to completely prevent complications, 2) antithrombin III consumption can blunt efficacy in some patients, 3) low or intermediate doses may be insufficient, and 4) there was clinically significant cross-reactivity of the pathogenic HIT antibodies. CONCLUSIONS: It is emphasized that the possibility of clinically significant antibody cross-reactivity and that low or intermediate dosage may be inadequate when using danaparoid in therapy of HIT. The latter problem probably extrapolates to other anticoagulants used for HIT. PMID- 12890150 TI - Allogeneic hematopoietic cell transplantation for paroxysmal nocturnal hemoglobinuria. AB - BACKGROUND: Although allogeneic hematopoietic cell transplantation (HCT) has a potential to cure patients with paroxysmal nocturnal hemoglobinuria (PNH), appropriate indication and conditioning regimen for HCT have not been established. PATIENTS AND METHODS: Between July 1999 and December 2001, five patients with PNH underwent allogeneic HCT: three for refractory hemolysis and two for aggravating cytopenia. Four patients with hypercellular marrow received Bu-Fludara-ATG (busulfan 4 mg/kg/d for 2 d, fludarabine 30 mg/m2/d for 6 d, and ATG 20 mg/kg/d for 4 d) for conditioning therapy and one patient with hypocellular marrow was conditioned with Cy-ATG (cyclophosphamide 50 mg/kg/d for 4 d and ATG 30 mg/kg/d for 3 d). Three patients received stem cell graft from matched sibling donor and two patients from 1-antigen mismatched unrelated donor. RESULTS: One patient who was conditioned with Bu-Fludara-ATG failed to engraft and died at post-transplant day 62. The other four patients showed three lineage engraftment and normal expression of CD55 and CD59 antigens by flow cytometric analysis. They are alive with stable engraftment and full donor chimerism between post-transplant day 510 and 1116. Acute graft vs. host disease (GVHD) of grade II or more occurred in two patients and extensive chronic GVHD in four. CONCLUSION: HCT using related or unrelated donor could eradicate PNH clones and may cure patients with the disease. Further studies are needed to establish the role of allogeneic HCT, especially with reduced intensity conditioning therapy, in the treatment of PNH. PMID- 12890151 TI - Quantitative determination of the human MRP1 and MRP2 mRNA expression in FACS sorted peripheral blood CD4+, CD8+, CD19+, and CD56+ cells. AB - OBJECTIVES: ATP-binding cassette (ABC) transporters extrude a wide variety of endogenous and exogenous compounds. In cancer cells, they are known to confer multidrug resistance. The aim of the present study was to determine the expression of the multidrug resistance-associated protein 1 (MRP1) and 2 (MRP2), which are members of the subfamily C of the ABC transporters family, in human hematopoietic cells. METHODS: CD4+, CD8+, CD19+, and CD56+ cells were isolated from whole blood by FACS-sort in 20 healthy volunteers. MRP1 and MRP2 mRNA levels were quantified using real-time reverse transcriptase-polymerase chain reaction (RT-PCR) assays on a LightCycler (Roche, Mannheim, Germany). RESULTS: The MRP1 mRNA exhibited the highest abundance in CD4+ cells (7.4 x 10(3)+/-3.19 x 10(3) molecules/ng of total RNA), followed by CD8+ > CD19+ > CD56+ cells. The MRP2 mRNA expression was highest in CD4+ cells (6.7 x 10(2)+/-2.84 x 10(2)), followed by CD8+ > CD56+ > CD19+ cells. No correlation between the MRP1 and MRP2 mRNA expression was observed. Interestingly, beta2-microglobulin mRNA expression in CD19+ cells was found to be twofold lower in comparison with other cells. CONCLUSIONS: On an mRNA level both MRP1 and MRP2 were expressed in peripheral blood cells, with more than sevenfold higher MRP1 expression in all cell populations investigated. The impact of the MRP1 and MRP2 transcription in these cells remains to study. The use of beta2-microglobulin as a housekeeping gene could have a critical impact on the interpretation of RT-PCR data. PMID- 12890152 TI - Granular lymphocyte proliferative disorder after autologous peripheral blood stem cell transplantation for multiple myeloma. AB - A 57-yr-old woman with multiple myeloma underwent tandem autologous peripheral blood stem cell transplantation (APBSCT). Fever, anemia, and thrombocytopenia, followed by the proliferation of granular lymphocytes in the peripheral blood occurred, after a second APBSCT. Clonal rearrangement of the T-cell receptor was detected using Southern blot analysis of peripheral blood samples. Granular lymphocyte proliferative disorders (GLPD) were diagnosed. After steroid therapy, the symptoms resolved. Lymphocytosis did not recur after the discontinuation of steroids. There have been a few reports of GLPD after solid organ and allogeneic hematopoietic stem cell transplantations. We report a first case of GLPD after APBSCT. PMID- 12890153 TI - Treatment of a patient with chronic renal failure with rituximab for a follicular lymphoma: safe and successful option of rituximab therapy. AB - A 47-yr-old woman presented a chronic renal failure for 5 yr, with a creatinine clearance of 12 mL/min. In June 2002, she had a right axillary lymph node (of 4 cm diameter). A biopsy revealed a follicular lymphoma (histology: follicular small cleaved-cell). She had Ann Arbor stage III disease, with a high tumor burden according to the GELF criteria. She received rituximab as single first line treatment (375 mg/m2 by intravenous infusion for a total of four dosages: days 1, 8, 15 and 22). Rituximab therapy was extremely well tolerated, and we obtained a partial response, 4 wk after completing the treatment. In January 2003, she received one maintenance course of rituximab. Six weeks after maintenance therapy, a complete response was achieved. PMID- 12890154 TI - Aplastic anaemia associated with a Philadelphia chromosome and monosomy 7 during immunosuppressive therapy. AB - We describe a patient who presented with aplastic anaemia associated with the Philadelphia (Ph1) chromosome during immunosuppressive therapy and who subsequently developed myelodysplastic syndrome (MDS) with monosomy 7. Initially the patient had hypocellular fatty marrow without leukaemic blasts or dysplastic features. Chromosome analysis showed 46, XY, t(9;22)(q34;q11) during immunosuppressive therapy, but no leukaemic transformation was detected. The patient showed gradual haematologic improvement and became transfusion independent. Thereafter, bone marrow dysplasia with monosomy 7 progressed following transfusion independence. These findings indicate that multiple cytogenetic evolutions occur in aplastic anaemia during immunosuppressive therapy, and that Ph1 chromosome may play a role in bone marrow suppression rather than development of leukaemia. PMID- 12890155 TI - Dinucleotide deletion in -alpha3.7 allele causes a severe form of alpha+ thalassaemia. AB - We describe a family of Italian origin in which the father and his two children had hypochromia and microcytosis with normal iron status. All individuals underwent an uneventful clinical course and required no treatment. To investigate the molecular basis of this phenotype, which is a prerequisite for further genetic counselling, we revealed that all affected family members are carriers of a common form of alpha+ thalassaemia resulting from the deletion of 3.7 kb of the alpha-globin cluster (alphaalpha/-alpha3.7). However, this genotype alone could not account for the phenotype presenting in this family. Further characterization of the alpha-globin genes demonstrated an additional AC deletion in the vicinity of the initiation codon of the -alpha3.7 allele. This secondary mutation causes an additional impaired translation of the affected allele producing increased globin chain imbalance. This leads to a more severe phenotype, as heterozygotes for such mutation (alphaalpha/-alphaT) have hypochromic microcytosis and abnormal globin chain synthesis that mimic alpha0 thalassaemia trait (--/alphaalpha). Accurate genotyping of alpha globin determinant is absolutely required as there is a possibility that an interaction of this unusual double mutation with other common alpha0 thalassaemias (--/-alphaT) can give rise to a very severe, probably fatal, alpha thalassaemia. PMID- 12890156 TI - Tryptase-positive mast cells predict clinical outcome of patients with early B cell chronic lymphocytic leukemia. PMID- 12890159 TI - The use of genetic microarray analysis to classify and predict prognosis in haematological malignancies. AB - The introduction of microarrays offers the opportunity to examine the expression of many thousands of genes in a single experiment. Investigations in leukaemia and lymphoma have led to the identification of a number of subgroups, with a defined gene expression pattern, not previously identified by morphology, cytogenetics or molecular techniques. In many cases these expression patterns can be linked to the tumour cells normal developmental counterpart, and represent distinct disease subgroups with different clinical presentations and outcomes. The technology has also identified genes that may be important in tumour cell biology including key genes in cell proliferation, adhesion, apoptosis, and the development of drug resistance. These early studies demonstrate that genetic microarrays will be useful in classifying haematological malignancies, predicting response to treatment, predicting prognosis, and identifying novel targets for therapy. PMID- 12890160 TI - Report on slide session, British Society for Haematology, 42nd annual scientific meeting, Brighton, 2002. PMID- 12890161 TI - Iron absorption from Spatone (a natural mineral water) for prevention of iron deficiency in pregnancy. AB - The absorption of iron from Spatone Iron-Plus has been investigated in pregnant women with iron deficiency anaemia. A total of 25 mg Fe was taken and absorption determined from the increase in serum iron concentration during a period of 3 h. Mean absorption was 28%, significantly higher than in nonpregnant, nonanaemic women (14%). These studies demonstrate that Spatone provides an alternative to the standard ferrous sulphate tablet for prevention of iron deficiency in pregnancy. As only lower doses of iron are required, the unpleasant side-effects of iron therapy are largely avoided. PMID- 12890162 TI - Overexpression of p16INK4A and p14ARF in haematological malignancies. AB - Two proteins, p16INK4A and p14ARF, originating from the same gene locus CDKN2A, use different promoters and alternative reading frames. p16INK4A is translated from alpha transcript and p14ARF is from beta transcript. These two proteins, which are inactivated in some human malignancies, are possible tumour suppressor candidates. In this study, we investigated the expression of p16INK4A and p14ARF mRNAs in haematological malignancies. We studied eight normal bone marrow samples, three reactive granulocytic hyperplasia patients, and 21 haematological malignancy patients, including seven acute myelogenous leukaemia, four acute lymphoblastic leukaemia, five myelodysplastic syndrome, five chronic myelogenous leukaemia (CML). p16INK4A and p14ARF mRNA expression was assayed by reverse transcriptase polymerase chain reaction. Normal bone marrows and reactive granulocytic hyperplasia showed barely detectable expression of either mRNA. In contrast, p16INK4A and p14ARF mRNA expression was abnormally increased in patients with haematological malignancies. Especially in CML, overexpression of p16INK4A and p14ARF mRNAs was more frequent than in controls (80 and 60%, respectively, P < 0.05). In conclusion, p16INK4A and p14ARF mRNA expression was frequently increased in haematological malignancies, especially in CML. We suggest that overexpression of these mRNAs may be related to the pathogenesis of haematological malignancies. PMID- 12890163 TI - Variable manifestation in natural killer cell leukaemia. AB - Natural killer (NK) cell leukaemias are a relatively rare group of haematological disorders, now entitled in the T/NK lymphoproliferative disorders in the new WHO classification. Recent studies have clarified their biological and clinical manifestation gradually. However, some cases with NK malignancies still remain difficult to diagnose and differentiate into their subtypes in the absence of a distinct diagnostic hallmark, especially at initial presentation. We describe herein five patients with NK leukaemias with respect to the clinical, cytological, immunological and cytogenetic characteristics, varied among each case. Cytologically, two aggressive NK cell leukaemia/lymphoma (ANKL/L) cases were a morphologically hypogranular variant form. Clinically, one with ANKL/L was presented as haemophagocytic syndrome without leukaemic infiltration. Systemic chemotherapy resulted in complete remission in one ANKL/L and two blastic NK cell leukaemia/lymphoma (BNKL/L) patients; however, a good long-term outcome was achieved in only one CD4-positive BNKL/L patient with allogenic bone marrow transplantation. Cytogenetic analysis revealed that recurrent chromosomal aberration was rare; however, two had aberrations at 10p11 and 11q13. From these findings, we conclude that comprehensive individual studies should be carried out in these patients to obtain a correct diagnosis and to design an optimal therapeutic approach. PMID- 12890164 TI - Ethnic variation in von Willebrand factor levels can influence the diagnosis of von Willebrand disease. AB - The amount and function of von Willebrand factor (VWF), measured against a panel of laboratory tests, is the normal basis for the diagnosis of von Willebrand's disease (VWD). The normal range for each test is usually obtained by assaying samples from a cross section of the local population or from a manufacturer's kit insert. While collecting normal controls for another study, population from Durban in South Africa, with its distinct ethnic mix of Africans, Indians and Caucasians were also studied. Previously, Indians from their subcontinent have not been looked at separately and compared with Africans and Caucasians. It is confirmed in a previous study (Miller, Dilley, Richardson, Craig, Evatt. (2001) American Journal of Hematology 67, 125) that African Americans had significantly higher VWF:Ag and FVIII levels when compared with Caucasians. In addition, it was found that there was a significant difference in VWF:Ag levels between Indians and Africans, and between Caucasians and Africans, whereas no significant difference between Indians and Caucasians. Africans, Indians and Caucasians with blood group O showed significantly lower VWF:Ag and FVIII than the other ABO blood groups. Normal ranges of VWF for different blood groups are well established and this information should be utilized while considering a diagnosis of VWD. It is proposed here that the influence of racial origin should also be considered in the clinical and laboratory evaluation of VWD. PMID- 12890165 TI - No alteration in platelet function or coagulation induced by EGb761 in a controlled study. AB - Some cases of spontaneous bleeding have been reported in patients treated with Ginkgo biloba. A prospective, double-blind, randomized, placebo-controlled study was carried out in 32 young male healthy volunteers to evaluate the effect of three doses of Ginkgo biloba extract (120, 240 and 480 mg/day for 14 days) on hemostasis, coagulation and fibrinolysis. This study did not reveal any alteration of platelet function or coagulation. This suggests that the reported clinical bleeding events in patients receiving Ginkgo biloba extract are not related to pharmacological properties of EGb761. PMID- 12890166 TI - A novel t(15;17) translocation in acute myeloid leukaemia not associated with PML/RARalpha rearrangement. AB - We report a novel t(15;17)(q15;q11) translocation in acute myeloid leukaemia (AML M4) which was not associated with PML/RARalpha rearrangement or with acute promyelocytic leukaemia (APL) morphology. The leukaemia behaved in refractory fashion, with 70% blasts in the bone marrow after the first course of chemotherapy. In view of the refractory behaviour of the leukaemia, the patient was treated with high dose chemotherapy and autologous stem cell rescue. The patient is alive and well 22 months post-autologous stem cell transplant. This case demonstrates that while cytogenetic analysis provides important diagnostic and prognostic information, the precise location of the chromosomal breakpoints is critical in the interpretation of cytogenetic results. PMID- 12890167 TI - Haemoglobin Etobicoke, an incidental finding in an Irish diabetic. AB - It is well recognized that haemoglobin variants can be detected during the measurement of HbA1c by high-performance liquid chromatography (HPLC). A number of variants have been reported as compromising the quantification of HbA1c, a marker used in the assessment of glycaemic control in diabetes. We describe a case of haemoglobin Etobicoke, a rare alpha chain variant detected in an Irish diabetic during HbA1c analysis. Its identity was confirmed using a series of investigations. These included haemoglobin electrophoresis at alkaline and acid pH, isoelectric focusing and globin chain electrophoresis. Ultimately mass spectrometry isolated the mutation at position alpha 84 (F5). Haemoglobin Etobicoke, first described in Canada in 1969 has not previously been detected on HbA1c analysis. In the presence of this rare variant, HbA1c, a standard method using HPLC to assess glycaemic control in diabetes is unreliable and alternatives such as fructosamine need to be considered. HbA1c measured by automated HPLC will effectively screen populations where haemoglobin variants were not previously known. Precise identity of these variants when they are detected is crucial to the reliable interpretation of HbA1c analyses. PMID- 12890168 TI - Spontaneous rupture of the spleen as the presenting feature of the blastoid variant of mantle cell lymphoma. AB - Spontaneous splenic rupture is rare, and particularly so as the initial presentation of a lymphoproliferative disorder. Although rare cases of splenic rupture have been reported in mantle cell lymphoma there has not been a report of the blastoid variant presenting in this manner. We report such a case in a 64 year-old man. PMID- 12890169 TI - Acute arterial thrombosis in acute promyelocytic leukaemia. AB - INTRODUCTION: Localized large vessel thrombosis in acute leukaemia is rare, haemorrhagic complications being more common. METHOD: We present a patient with acute promyelocytic leukaemia (APL) presenting with an acutely ischaemic lower limb. Large vessel thrombosis is a rare presentation of APL. We reviewed the literature on the coagulopathy of APL and discuss the pathology and current treatment options. DISCUSSION: Disordered haemostasis is typical of acute promyelocytic leukaemia (FAB M3) and relates to the intrinsic properties of the blast cells as well as thrombocytopenia from bone marrow involvement. Expression of procoagulants, stimulation of cytokines and alterations in endothelial cell anticoagulant properties initiate a disseminated intravascular coagulation (DIC) resulting in the typical clinical and laboratory findings in APL. The promyelocytes are characterized by the balanced reciprocal translocation between chromosomes 15 and 17. All-trans-retinoic acid (ATRA) induces differentiation in these cells, revolutionizing the treatment of APL. CONCLUSION: Unexpected limb ischaemia in a young, apparently healthy patient might be the presenting symptom of an underlying haematological disorder such as APL. A thorough haematological investigation should be performed prior to contemplating surgery. New treatment strategies based on knowledge of the molecular biology of APL has improved the prognosis of patients suffering from APL. PMID- 12890170 TI - Spuriously elevated inorganic phosphate level in a multiple myeloma patient. AB - We report the case of a patient with IgG multiple myeloma and pseudohyperphosphatemia. The patient had no clinical features of hyperphosphatemia. Subsequent investigations demonstrated that this hyperphosphatemia was spurious and was caused by a high concentration of the paraprotein. Deproteinization of the serum samples by sulfosalicylic acid resulted in normalization of the elevated phosphate values. This pseudohyperphosphatemia resulted from an increase in optic density because of interference between monoclonal immunoglobulin and the molybdic reagent used to determine phosphate in serum. These data indicate that the finding of marked hyperphosphatemia in multiple myeloma patients should always prompt an assay carried out on a deproteinized sample. In addition, knowledge of this phenomenon may avoid confusion, unnecessary testing and obviate confusion in the clinical evaluation of patients with multiple myeloma. PMID- 12890171 TI - Cardiac hypertrophy: a matter of translation. AB - 1. Left ventricular hypertrophy (LVH) of the heart is an adaptive response to sustained increases in blood pressure and hormone imbalances. Left ventricular hypertrophy is associated with programmed responses at the molecular and biochemical level in different subsets of cardiac cells, including the cardiac muscle cells (cardiomyocytes), fibroblasts, conductive tissue and coronary vasculature. 2. Regardless of the initiating cause, the actual increase in chamber enlargement is, in each case, due to an increase in size of a pre existing cardiomyocyte population, with little or no change in their number; a process referred to as cellular hypertrophy. 3. An accelerated rate of global protein synthesis is the primary mechanism by which protein accumulation increases during cardiomyocyte hypertrophy. In turn, increased rates of synthesis are a result of increased translational rates of existing ribosomes (translational efficiency) and/or synthesis and recruitment of additional ribosomes (translational capacity). 4. The present review examines the relative importance of translational capacity and translational efficiency in the response of myocytes to acute and chronic demands for increased protein synthesis and the role of these mechanisms in the development of LVH. PMID- 12890172 TI - Effect of nitroglycerine in popliteal preparations from patients with peripheral occlusive arteriopathy precontracted with KCl or 5-hydroxytryptamine. AB - 1. At the present time, there are no studies in isolated arteries from patients suffering from peripheral occlusive arteriopathy (POA). In the present study, we attempt to characterize the effect of nitroglycerine (GTN) in isolated popliteal preparations obtained after leg amputation in 60-90-year-old men and women suffering from POA. 2. After surgical operation, arterial samples were stored in a refrigerator at 4 degrees C and, after 12-36 h, they were cut into rings and mounted in organ baths containing Krebs'-Henseleit solution at 37 degrees C and gassed constantly with 95% CO2 and 5% O2. Because noradrenaline elicited very poor contractile responses in these preparations, in the present study we evaluated the concentration-dependent contractions induced by KCl (15-90 mmol/L) and 5-hydroxytryptamine (5-HT; 10-7 to 10-4 mol/L) in arteriopathic popliteal rings and, when the corresponding maximum contractile effect had been obtained, we also evaluated the concentration-dependent relaxing effect produced by GTN (10 10 to 10-5 mol/L) in all precontracted preparations. As a reference, similar experiments were performed in popliteal preparations obtained following surgery on non-arteriopathic vascular tissue where it was necessary to resect a certain percentage of healthy vessel. 3. The responses to KCl and 5-HT were greater in healthy vessel than in arteriopathic rings. The relaxing effect of GTN was greater in preparations precontracted with 5-HT than in those preparations precontracted with KCl. In addition, preparations precontracted with KCl relaxed even less when they were obtained from patients with POA. 4. The present data indicate that GTN is a vasodilator with little effect on depolarized arteries. The results also indicate that the effect of this drug is even less in depolarized arteries from patients with POA. PMID- 12890173 TI - Hypocholesterolaemic effects of an ethanol precipitate of Kabosu juice in stroke prone spontaneously hypertensive rats fed a cholesterol-free diet. AB - 1. We have previously identified strong inhibitory effects of Kabosu (Citrus sphaerocarpa Hort.) juice precipitate (KJP) on cholesterol elevation in stroke prone spontaneously hypertensive rats (SHRSP) fed a cholesterol diet. In the present study, to elucidate the hypocholesterolaemic mechanism, we examined the effect of dietary KJP on lipid metabolism by using SHRSP fed a cholesterol-free diet. 2. Compositions of the experimental diet containing 10% KJP powder were adjusted to those of the control diet. Seven-week-old male SHRSP were fed control or experimental diet for 2 weeks with free access to the diet and water. 3. Serum levels of cholesterol, phospholipid and triglyceride of the KJP group were significantly reduced, which was due to decreases in the very low-density lipoprotein (VLDL), low-density lipoprotein (LDL) and high-density lipoprotein (HDL) fractions. 4. Serum concentrations of apolipoproteins A-I and E (apoA-I and E) of the KJP group were significantly lower than those of the control group, whereas no significant differences were observed in serum apoB and apoA-IV between the two groups. 5. In liver, there were no significant differences in the contents of lipids or relative liver weight between the two groups. The activity of microsomal cholesterol 7alpha-hydroxylase of the KJP group tended to increase, whereas that of microsomal acyl-coenzyme A : cholesterol acyltransferase was significantly reduced, compared with the control group. 6. These results indicate that dietary KJP produces reductions of serum lipid levels, which are due to reductions in VLDL, apoE HDL and apoA-I HDL, and may promote catabolism and excretion of hepatic cholesterol in SHRSP fed a cholesterol-free diet. PMID- 12890174 TI - Biochemical and morphological characterization of spontaneously hypertensive hyperlipidaemic rats. AB - 1. To develop and characterize a new animal model of hypertension and hyperlipidaemia, we cross-bred spontaneously hypertensive rats (SHR) with spontaneously hyperlipidaemic rats (HLR). 2. A new strain of spontaneously hypertensive hyperlipidaemic rats (SHHR) was established at generation 10 through selective mating of brothers and sisters (systolic blood pressure > 150 mmHg, plasma cholesterol > 150 mg/dL). Cross-bred Wistar-Kyoto (WKY) rats and Sprague Dawley (SD) rats (SDWKY rats) were used as a control. 3. Adrenaline and noradrenaline levels in the plasma and adrenal medulla of male SHHR were significantly higher than those of male SDWKY rats. The tyrosine hydroxylase activity in the adrenal medulla of male SHHR was significantly higher than that of male SDWKY rats. Low-density lipoprotein expression was found in the plasma of male and female SHHR and HLR. Cholesterol 7alpha-hydroxylase mRNA expression in the liver of male SHHR was lower than that of male SDWKY rats. Endothelium lesions and lipid deposition under the endothelium were observed in the aorta of 24-month-old SHHR, especially female SHHR, but not in age-matched HLR and SDWKY rats. 4. The hypertension of this new animal model of hypertension and hyperlipidaemia may be related to increased catecholamine activity and the hyperlipidaemia may be related to changes in the expression of cholesterol 7alpha hydroxylase mRNA and lipoprotein profiles. The SHHR may be valuable in the study of mechanisms of atherosclerosis and the evaluation of anti-atherosclerosis drugs as a new strain of cardiovascular disease. PMID- 12890175 TI - Protective effect of coadministered superoxide dismutase and catalase against stress-induced gastric mucosal lesions. AB - 1. There are conflicting reports as to the protective effect of coadministered native superoxide dismutase (SOD) and catalase against gastric mucosal lesions in rats with water immersion restraint (WIR) stress. It is unclear how coadministered native SOD and catalase protect against WIR stress-induced gastric mucosal lesions. Therefore, in the present study, we re-examined the protective effect of coadministered native SOD and catalase against gastric mucosal lesions in rats with WIR stress. 2. Gastric mucosal lesions were induced in Wistar rats by 3 h WIR. Rats were injected subcutaneously with a mixture of purified bovine erythrocyte SOD and bovine liver catalase 1 h before the onset of WIR. Ulcer index, serum SOD, catalase and xanthine oxidase (XO), uric acid and gastric mucosal SOD, catalase, XO, myeloperoxidase (MPO; an index of tissue neutrophil infiltration), non-protein sulfhydryl (NP-SH) and thiobarbituric acid-reactive substances (TBARS; an index of lipid peroxidation) were assayed in all rats used. 3. Rats with 3 h WIR showed gastric mucosal lesions. Pre-administration of SOD plus catalase to rats with WIR prevented lesion formation. In the serum of rats with WIR alone, XO activity and uric acid concentration increased, whereas SOD and catalase activities did not change. Pre-administration of SOD plus catalase to rats with WIR did not affect increased serum XO activity and uric acid concentration, but did increase serum SOD and catalase activities. In the gastric mucosa of rats with WIR alone, increases in MPO activity and TBARS concentration and a decrease in NP-SH concentration occurred, whereas XO, SOD and catalase activities did not change. Pre-administration of SOD plus catalase to rats with WIR attenuated the changes in gastric mucosal MPO activity and TBARS and NP-SH concentrations, but did not affect gastric mucosal XO, SOD and catalase activities. Pre-administration of SOD plus catalase (in an inactivated form) to rats with WIR had no effect on gastric mucosal lesion formation and the levels of serum and gastric mucosal parameters studied. 4. These results indicate that coadministered native SOD and catalase protect against gastric mucosal lesions in rats with WIR stress and suggest that this protective effect of coadministered native SOD and catalase could be due to their activity to scavenge XO-derived active oxygen species that are increased in the blood. PMID- 12890176 TI - Effects of chronic hypobaric hypoxia on contractile properties of rat sternohyoid and diaphragm muscles. AB - 1. Chronic hypoxia occurs in a variety of circumstances, including respiratory disease and exposure to altitude, and is known to affect respiratory muscle structure. However, little is known about its effects on respiratory muscle contractile properties. 2. Rats were exposed to normoxia (n = 16) or hypobaric hypoxia (n = 16; barometric pressure 450 mmHg) for 6 weeks. Contractile properties were measured in isolated sternohyoid and diaphragm muscles in warmed, oxygenated Krebs' solution. Isometric twitch and tetanic tension, contraction time, half-relaxation time and tension-frequency relationship were determined using field stimulation with platinum electrodes. Fatigue was induced by stimulation at 40 Hz with 300 msec trains of 0.5 Hz for 5 min. 3. Chronic hypoxia had no effect on bodyweight, but did increase haematocrit. Chronic hypoxia increased specific force development in both muscles and increased sternohyoid fatigue. Chronic hypoxia had no effect on contractile kinetics in either muscle, but shifted the tension-frequency relationship to the left in the diaphragm. 4. Therefore, chronic hypoxia alters rat respiratory muscle force and fatigue, either due to the direct effects of hypoxia or to increased muscle activation. PMID- 12890177 TI - Effects of preweaning doxazosin treatment on adult pressure in the spontaneously hypertensive rat. AB - 1. The neonatal/preweaning period appears to represent a critical period of involvement of the sympathetic nervous system in the development of hypertension in spontaneously hypertensive rats (SHR). 2. We tested whether alpha1 adrenoceptor-mediated effects during the preweaning period are involved in the development of hypertension in the adult SHR. 3. Male SHR were treated with the alpha1-adrenoceptor antagonist doxazosin (10 mg/kg per day, s.c.) from postnatal day 1 to 21 inclusive. Direct conscious blood pressure and heart rate were measured via the caudal artery at 12 weeks of age. 4. Preweaning treatment with doxazosin had no significant effect on mean arterial blood pressure or heart rate in male SHR at 12 weeks of age. 5. These findings do not support the involvement of alpha1-adrenoceptor-mediated effects during the preweaning period in the development of hypertension in adult SHR. PMID- 12890179 TI - Membranes as metabolic pacemakers. AB - 1. In the present review, we suggest that a few common processes linked to membranes consume the majority of energy used by most organisms. 2. Membranes may act as metabolic pacemakers through changes in lipid composition, altering membrane characteristics and the working environment of membrane proteins. 3. Experiments involving membrane exchanges show predictable changes in protein activities (sodium pump) that are dependent upon the type of membrane used. 4. Potential mechanisms discussed include fluidity, electrical fields, surface area requirements of lipids and peptide-lipid interactions. PMID- 12890180 TI - Energy cost of contraction in rat urinary bladder smooth muscle during anoxia. AB - 1. The aim of the present study was to investigate the effects of hypoxia on energy metabolism and contraction of rat urinary bladder smooth muscle, thereby gaining insight into the capacity of this smooth muscle to maintain contractile function when rendered hypoxic. 2. Isometric force, oxygen consumption, lactate production, heat production and unloaded shortening velocity were measured in isolated muscle strips under both aerobic and anaerobic conditions. Muscle strips were bathed in physiological saline solution with the anaerobic condition being created by replacing the oxygen bubbling the solution with nitrogen. 3. During contraction under anaerobic conditions, the rate of lactate production was increased 2.5-fold above that observed under aerobic conditions. This, however, only provided for a rate of ATP production of approximately 30% of that measured under aerobic conditions. Despite this, force maintenance was only slightly depressed, indicating that the metabolic cost of contraction was reduced in hypoxia. In support of this, the rate of heat production during contractions in anoxia was only approximately half of that under aerobic conditions, whereas, again, force was only slightly lower. Unloaded shortening velocity was significantly lower in anoxia, suggesting a slower cross-bridge turnover rate. 4. The results indicate that the economy of force maintenance is increased in bladder smooth muscle under hypoxic conditions and that this is due, at least in part, to a reduced rate of cross-bridge cycling. This may help to preserve bladder contractile function during periods of ischaemia that may be associated with bladder filling and emptying. PMID- 12890181 TI - Relationship between isometric force and myofibrillar MgATPase at short sarcomere length in skeletal and cardiac muscle and its relevance to the concept of activation heat. AB - 1. This paper has been written in recognition of the seminal contributions to cardiac and skeletal muscle energetics made by Professor Colin Gibbs during his distinguished academic career. 2. The paper focuses on what is now known about the relationship between Ca2+-activated isometric force production and myofibrillar MgATPase in intact and skinned (surface membrane rendered permeable) skeletal and cardiac muscle preparations at short sarcomere lengths. 3. The relevance of this relationship to understanding the interactions between the actin and myosin filaments at the cross-bridge level in the region of double actin filament overlap and the cellular basis of 'activation heat' measurements in intact striated muscles is discussed. PMID- 12890182 TI - Force-matching errors after eccentric exercise attributed to muscle soreness. AB - 1. Experiments were performed to test the ability of human subjects to match forces in their elbow flexor muscles following eccentric exercise of one arm and, in a second series, after biceps brachii of one arm had been made sore by injection of hypertonic saline. 2. In the force-matching task, the elbow flexors of one arm, the reference arm, generated 30% of maximum voluntary contraction (MVC) under visual control. Subjects matched that level with their other arm, the indicator arm, without visual feedback. 3. After eccentric exercise of elbow flexors of the indicator arm, subjects felt they had achieved a satisfactory match while indicating forces that were significantly lower, by approximately 5%, than the reference level. Errors were in the opposite direction (i.e. forces were overestimated) when the reference arm was exercised. 4. Errors were reduced when matching forces were expressed as fractions of the sessional MVC rather than the pre-exercise MVC. Residual errors from 24 h postexercise onwards were attributed to muscle soreness from the exercise. 5. In support of this view, a similar pattern of matching errors was observed when an unexercised arm was made sore by injection of hypertonic saline into the biceps. 6. It is concluded that muscle soreness can interfere with a subject's ability to match forces, perhaps as a result of a reduced excitability of motor cortex. It implies that muscle soreness may contribute to the weakness experienced after a period of unaccustomed eccentric exercise. PMID- 12890183 TI - Cardiac energetics: from E(max) to pressure-volume area. AB - 1. To celebrate this Festschrift for Professor Colin Gibbs, as an invited speaker, I would like to review briefly my 35 year research career in cardiac physiology. 2. My career started in the late 1960s in Tokyo with my serendipitous discovery of Emax (ventricular end-systolic maximum elastance) as a load independent contractility index based on the time-varying elastance (E(t)) model of the ventricle. Professor K Sagawa at the Johns Hopkins University, USA, whom I joined in 1971, encouraged me to go further. 3. The next serendipitous event in my career was the discovery of ventricular pressure-volume area (PVA) as a measure of total mechanical energy of ventricular contraction in the late 1970s. The PVA concept was theoretically deducible from the E(t) and Emax concept and correlated surprisingly well with ventricular O2 consumption (Vo2). 4. Professor Gibbs' intuitive recognition of the significance of PVA in myocardial energetics in the 1980-1990s greatly encouraged me thereafter. The third serendipitous event in my career occurred in the mid 1990s and was my discovery of a novel integrative analysis method to assess the total amount of Ca2+ recruited in each excitation-contraction coupling from the decay rate of postextrasystolic potentiation, taking advantage of the Emax-PVA-Vo2 framework. 5. I am now hoping to experience one more serendipitous experience by developing an integrative analysis method of cross-bridge cycling in a beating heart using the Emax and PVA concepts. PMID- 12890184 TI - Species-independent metabolic response to an increase of [Ca(2+)](i) in quiescent cardiac muscle. AB - 1. The aim of the present investigation was to contrast the Ca2+ dependence of cardiac energy metabolism in two species with differential reliance on extracellular Ca2+ for excitation-contraction coupling. 2. We measured energy expenditure as the rate of oxygen consumption (Vo2) of isolated, Langendorff perfused hearts of rats and guinea-pigs during KCl arrest. In parallel experiments, we indexed intracellular Ca2+ concentration ([Ca2+]i) of isolated right-ventricular trabeculae, using the Ca2+ fluorophore fura-2 and ratiometric spectrofluorometry. By varying extracellular Na+ concentration ([Na+]o), Vo2 [Na+]o and [Ca2+]i-[Na+]o relationships were constructed for each species. 3. Reduction of [Na+]o during K+ arrest caused pronounced species-dependent elevations of both Vo2 and [Ca2+]i. Despite the species dependence of both Vo2 and [Ca2+]i on [Na+]o, a single species-independent Vo2-[Ca2+]i relationship obtained. 4. We infer that elevation of the metabolic rate of the arrested heart above its basal value is determined primarily by [Ca2+]i and is not species dependent. PMID- 12890186 TI - Cardiac energetics: sense and nonsense. AB - 1. The background to current ideas in cardiac energetics is outlined and, in the genomic era, the need is stressed for detailed knowledge of mouse heart mechanics and energetics. 2. The mouse heart is clearly different to the rat in terms of its excitation-contraction (EC) coupling and the common assumption that heart rate difference between mice and humans will account for the eightfold difference in myocardial oxygen consumption is wrong, because the energy per beat of the mouse heart is approximately one-third that of the human heart. 3. In vivo evidence suggests that there may well be an eightfold species difference in the non-beating metabolism of mice and human hearts. It is speculated that the magnitude of basal metabolism in the heart is regulatable and that, in the absence of perfusion, it falls to approximately one-quarter of its in vivo rate and that in clinical conditions, such as hibernation, it probably decreases; its magnitude may be controlled by the endothelium. 4. The active energy balance sheet is briefly discussed and it is suggested that the activation heat accounts for 20-25% of the active energy per beat and cross-bridge turnover accounts for the balance. It is argued that force, not shortening, is the major determinant of cardiac energy usage. 5. The outcome of recent cardiac modelling with variants of the Huxley and Hill/Eisenberg models is described. It has been necessary to invoke 'loose coupling' to replicate the low cardiac energy flux measured at low afterloads (medium to high velocities of shortening). 6. Lastly, some of the unexplained or 'nonsense' energetic data are outlined and eight unsolved problems in cardiac energetics are discussed. PMID- 12890185 TI - On being the right size: heart design, mitochondrial efficiency and lifespan potential. AB - 1. From the smallest shrew or bumble-bee bat to the largest blue whale, heart size varies by over seven orders of magnitude (from 12 mg to 600 kg). This study reviews the scaling relationships between heart design, cellular bioenergetics and mitochondrial efficiencies in mammals of different body sizes. 2. The [31P] nuclear magnetic resonance-derived [phosphocreatine]/[ATP] ratio in hearts of smaller mammals is significantly higher (2.7 +/- 0.3 for mouse; n = 22) than in larger mammals (1.6 +/- 0.3 for humans; n = 13). 3. The inverse of the free myocardial cytosolic [ADP] concentration and the cytosolic phosphorylation ratio ([ATP]/[ADP][Pi]) scales with heart size and with absolute mitochondrial and myofibrillar volumes, close to a quarter-power (from -0.22 to -0.28; r = 0.99). 4. Assuming a similar mitochondrial P/O ratio and the same maximal amount of work required to convert 1 mol NADH to 0.5 mol O2 (i.e. 212.25 kJ/mol), the higher [ATP]/[ADP][Pi] ratios or cellular driving forces (DeltaG'ATP) in hearts of smaller mammals imply greater mitochondrial efficiencies in coupling ATP production to electron transport as body size decreases. For a P/O ratio of 2.5, the mitochondrial efficiency in the heart of a shrew, mouse, human and whale is 84, 82, 71 and 65%, respectively. 5. Higher cytosolic ATP]/[ADP][Pi] ratios and DeltaG'ATP values imply that the hearts of smaller mammals operate further from equilibrium than hearts of larger mammals. 6. As a consequence of scaling relationships, a number of remarkable invariants emerge when comparing heart function from the smallest shrew to the largest whale; the total volume of blood pumped by each heart in a lifetime is approximately 200 million L/kg heart and the total number of heart beats is approximately 1.1 billion per lifetime. 7. Similarly, the metabolic potential (total O2 consumed during adult lifespan per g bodyweight) for a 2 g shrew or a 100000 kg blue whale is approximately 38 L O2 consumed or 8.5 mol ATP/g body mass per lifetime. 8. The importance of quarter power scaling relationships linking structural, metabolic and bioenergetic design to the natural ageing process and maximum lifespan potential is discussed. PMID- 12890188 TI - Surgical treatment of melanoma. PMID- 12890189 TI - Elevated matrix metalloproteinase-2 and -3 production from human diabetic dermal fibroblasts. AB - BACKGROUND: Diabetic foot ulcers are characterized by elevated levels of matrix metalloproteinases (MMP), which could lead to excessive matrix breakdown and disruption to healing. It is unknown if this elevation is a function of wound healing, or if it is present within normal skin and a primary contributor to the increased risk of impaired healing. OBJECTIVES: To determine whether diabetic fibroblasts from unwounded skin show elevated MMP production compared with their nondiabetic counterparts. PATIENTS AND METHODS: Circular skin biopsies (4 mm diameter) were taken from the inside upper arm of four controls without diabetes and from four subjects with insulin-treated diabetes. Fibroblasts were incubated for a further 72 h and conditioned medium was collected and stored at -20 degrees C. The conditioned medium was assessed by gelatin zymography and Western blotting for MMP-2 and MMP-3. RESULTS: Diabetic dermal fibroblasts showed significantly elevated production of MMP-2 (P < 0.05) and pro-MMP-3 (P < 0.05) when compared with their nondiabetic counterparts. CONCLUSIONS: Dermal fibroblasts from normal unwounded skin are characterized by increased MMP production and this may be a primary contributing factor to the increased risk of nonhealing foot ulceration in diabetes. PMID- 12890190 TI - Involvement of histamine H3 receptors in scratching behaviour in mast cell deficient mice. AB - BACKGROUND: Although the roles of histamine H3 receptors have been studied in several tissues such as the brain, lung, spleen, colon and peripheral sensory nerve endings, the involvement of H3 receptors in skin responses particularly in relation to scratching behaviour are not well documented. OBJECTIVES: This work was performed to study the effects of histamine H3 antagonists on scratching behaviour in mast cell-deficient mice. METHODS: Histamine H3 antagonists iodophenpropit and clobenpropit, histamine and substance P were injected intradermally into the rostral part of the back of mast cell-deficient (WBB6F1 W/Wv) and wild-type (WBB6F1+/+) mice and scratching behaviour was measured for 60 min. The effects of H1 antagonists on scratching behaviour induced by H3 antagonists were also investigated. RESULTS: Intradermal injection of iodophenpropit and clobenpropit at doses of 10 and 100 nmol per site caused significant increases in scratching behaviour in both mast cell-deficient and wild-type mice. Histamine also caused a dose-related increase in the incidence of scratching behaviour, and a significant effect was observed at a dose of 100 nmol per site in both mast cell-deficient and wild-type mice. Substance P was also effective in causing scratching behaviour in both mast cell-deficient and wild type mice. However, histamine H1 antagonists diphenhydramine and chlorphenamine failed to inhibit H3 antagonist-induced scratching behaviour in both types of mice. CONCLUSIONS: Our results indicated that intradermal injection of H3 antagonists induces scratching behaviour and that chemical mediators other than histamine seem to be involved in the response. PMID- 12890191 TI - Morphological and biochemical studies of human beta-mannosidosis: identification of a novel beta-mannosidase gene mutation. AB - BACKGROUND: There are seven well-known lysosomal storage diseases that produce angiokeratoma corporis diffusum clinically. beta-Mannosidosis (MANB1; OMIM248510), first reported in humans in 1986, is a rare hereditary lysosomal storage disease caused by a deficiency of the enzyme beta-mannosidase. Since then, 13 cases of beta-mannosidase deficiency in ten families have been described. A human beta-mannosidase mutation has been reported only by Alkhayat et al. in 1998. OBJECTIVES: To clarify its pathogenesis we did electron microscopic, biochemical and molecular biological investigations of a Japanese patient with beta-mannosidosis. METHODS: Ultrastructural analyses, enzyme assays, cell culture and mRNA and genomic DNA were sequenced to find mutations in the beta-mannosidase gene. RESULTS: Electron microscopy of skin biopsy specimens from the patient showed cytoplasmic vacuolation of lysosomes in blood and lymph vessels, endothelial cells, fibroblasts, secretory portions of eccrine sweat glands, neural cells and basal keratinocytes in the epidermis. This vacuolation was also observed in cultured keratinocytes and fibroblasts. Assays of seven enzyme activities in plasma and cultured skin fibroblasts showed a marked decrease of beta-mannosidase activity. Sequencing the beta-mannosidase cDNA revealed a four-base (ATAA) insertion between exons 7 and 8, resulting in a frameshift at codon 321 and termination at codon 325. Analysis of the patient's genomic DNA revealed a novel homozygous A(+1)-->G splice site mutation in intron 7. CONCLUSIONS: To our knowledge, this is the first case of beta-mannosidosis reported in Japan and the second report in which a gene mutation is identified. The biological importance of beta-mannose moieties in glycoproteins in basal keratinocytes is suggested. PMID- 12890192 TI - Methotrexate inhibits proliferation and regulation of the expression of intercellular adhesion molecule-1 and vascular cell adhesion molecule-1 by cultured human umbilical vein endothelial cells. AB - BACKGROUND: The mechanism by which a low dose of methotrexate (MTX) works to treat psoriasis is not clear. The overexpression of cell adhesion molecules on dermal vessels is important in the pathogenesis of psoriasis and is probably induced by upregulation of tumour necrosis factor (TNF)-alpha. OBJECTIVES: To determine the effects of MTX at concentrations comparable with in vivo levels after the administration of low-dose MTX to human umbilical vein endothelial cells (HUVEC) on the growth and expression of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1). METHODS: Cell proliferation assay, immunostaining, immunoblotting, cell enzyme-linked immunosorbent assay and reverse transcriptase-polymerase chain reaction (RT-PCR) were performed to examine the effects of MTX on HUVEC. RESULTS: MTX inhibited the proliferation of HUVEC at 10-7 mol L-1 and 10-6 mol L-1 without showing cytotoxic effects. It also inhibited TNF-alpha-induced ICAM-1 and VCAM-1 expression by HUVEC at 10-6 mol L-1. The inhibitory effect of MTX was more pronounced on ICAM-1 expression than on VCAM-1 expression. RT-PCR analysis revealed that TNF-alpha induced ICAM-1 gene expression was strongly downregulated by MTX. CONCLUSIONS: Low-dose MTX may act on psoriasis by suppressing the TNF-alpha-induced expression of ICAM-1 and VCAM-1 by vascular endothelial cells. Inhibition of neovascularization may be another mechanism of action of MTX. PMID- 12890193 TI - Administration of anti-interleukin 18 antibody fails to inhibit development of dermatitis in atopic dermatitis-model mice NC/Nga. AB - BACKGROUND: Interleukin (IL)-18 has been shown to activate basophils to produce histamine and IL-4 and to induce naive T cells to differentiate into T-helper (Th) 2 cells. However, when expressed together with IL-12, IL-18 induces Th1 cell development and inhibits IgE synthesis. Previously we reported that serum IL-18 levels were elevated in the sera from atopic dermatitis-model mice NC/Nga, prior to the onset and during the development of dermatitis. OBJECTIVES: We studied whether neutralization of IL-18 activity might affect dermatitis in NC/Nga mice, to investigate the role of IL-18 on dermatitis. METHODS: NC/Nga mice were given weekly anti-IL-18 antibody starting at 5 weeks of age to 13 weeks and development of dermatitis, scratching behaviour and serum IgE concentrations were evaluated. RESULTS: Continuous injections of anti-IL-18 antibody failed to inhibit the onset and development of dermatitis and IgE elevation. The treatment, rather, tended to lead to an exacerbation of dermatitis and scratching behaviour. In addition, the administration of anti-IL-18 antibody did not ameliorate the responsiveness of lymphocytes to IL-4, which was previously demonstrated as an immunological abnormality in the mouse. CONCLUSION: This study demonstrates that, at least in NC/Nga mice, IL-18, although excessively expressed before the onset of dermatitis, shows antiallergic actions. PMID- 12890194 TI - Generation and characterization of epidermolysis bullosa simplex cell lines: scratch assays show faster migration with disruptive keratin mutations. AB - BACKGROUND: Epidermolysis bullosa simplex (EBS) is an inherited skin fragility disorder caused by mutations in keratin intermediate filament proteins. While discoveries of these mutations have increased understanding of the role of keratins and other intermediate filaments in epithelial tissues, progress towards the development of therapy for these disorders is much slower. OBJECTIVES: Cell culture model systems that display these structural defects are needed for analysis of the cellular consequences of the mutations and to enable possible therapeutic strategies to be developed. Our aim was to generate immortalized cell lines as such model systems for the study of EBS. METHODS: We generated a series of stable cell lines expressing EBS-associated keratin mutations, by immortalizing keratinocytes from EBS-affected skin biopsies with either simian virus 40 (SV40) T antigen or human papillomavirus 16 (HPV16) E6/E7, and assessed their keratin expression (by immunofluorescence), proliferation rates and migratory behaviour (in outgrowth and scratch wound assays). RESULTS: Clonal immortalized keratinocyte cell lines KEB-1, KEB-2, KEB-3 (using SV40 T antigen) and KEB-4, KEB-7 and NEB-1 (using HPV16 E6/E7) were established. These include two lines from a single individual with Weber-Cockayne EBS (i.e. KEB-3 and KEB-4, mutation K14 V270M), and three cell lines from a second family, two from siblings carrying the same mutation (KEB-1, KEB-2 lines from Dowling-Meara EBS, mutation K5 E475G) and one from an unaffected relative (NEB-1). The sixth cell line (KEB 7), with a previously unreported severe mutation (K14 R125P), was the only one to show keratin aggregates in resting conditions. Despite variations in the immortalization procedure, there was no significant difference between cell lines in keratin expression, outgrowth capabilities or response to transient heat shock. However, cell migration, as measured by speed of scratch wound closure, was significantly faster in cells with severe EBS mutations. CONCLUSIONS: These cell lines provide useful culture systems in which to assess aspects of EBS induced cell changes. The faster migration after scratch wounding of the EBS keratinocytes may be a consequence of the known upregulation of stress-activated kinase pathways in these cells. PMID- 12890195 TI - An evaluation of dermoscopy fluids and application techniques. AB - BACKGROUND: Dermoscopy, a noninvasive technique used to help physicians better visualize pigmented skin lesions, is becoming widely used by dermatologists. Yet despite its popularity, to our knowledge basic aspects such as the best immersion fluid (IF) to use and proper procedures for applying the IF and dermatoscope have never been the subject of a systematic investigation. OBJECTIVES: To determine the best techniques for application of IF and the dermatoscope; to discover which IF results in the least amount of air inclusions; to determine which IF provides the best image quality. MATERIALS AND METHODS: This study was divided into three phases. Phase I examined several techniques of taking digital dermoscopic photographs in an attempt to eliminate air bubbles. Phase II tested seven IFs to analyse quantitatively the number of air inclusions for each IF. Phase III investigated these same IFs to judge the image quality for each. RESULTS: Phase I: in our hands, the best technique to apply an IF is to use an eyedropper and place the IF directly onto the skin in combination with a 'roll-on technique': the dermatoscope's edge is placed on the skin first and is then rotated until the glass plate lies flat against the skin. Phase II: the alcoholic IF resulted in the least amount of air bubble inclusions (70% ethanol, 90% isopropanol and alcoholic disinfectant), while liquid paraffin (a mineral oil) resulted in the most air bubbles. Phase III: dermoscopic structures were equally clear with alcohols and liquid paraffin, but slightly blurry with ultrasound gel and water. CONCLUSIONS: Although 90% isopropanol performed slightly better in terms of air inclusions, we prefer to use 70% ethanol in most circumstances because it has the advantage of being odourless, unlike its counterpart. Additionally, ethanol will not stain clothing, does not crystallize on the dermatoscope, disinfects and evaporates immediately (does not need to be wiped off). For certain local considerations, such as dermoscopy on mucosa or the nail, we prefer to use ultrasound gel because it will not flow. PMID- 12890196 TI - A broad spectrum of human papillomavirus types is present in the skin of Australian patients with non-melanoma skin cancers and solar keratosis. AB - BACKGROUND: Human papillomavirus (HPV) may play a role in the pathogenesis of non melanoma skin cancer (NMSC) in epidermodysplasia verruciformis (EV) patients, but in the general population no specific HPV types have been associated with these lesions. Objectives To examine the spectrum of HPV types present in the skin and tumours of Australian patients with NMSC or solar keratosis (SK). METHODS: Biopsies from tumours, and cotton swab samples of perilesional skin and buttock skin from each of 59 Australian patients with basal cell carcinoma (BCC), squamous cell carcinoma (SCC) or SK were tested for HPV DNA by polymerase chain reaction (PCR) using HPV consensus (FAP) primers and by type-specific primers for HPV 38 and candidate HPV 92. The identification of HPV type from consensus PCR was performed by sequencing and comparison with GenBank. RESULTS: In total, 49 of 59 (83%) patients harboured HPV DNA, which was detected in 28 of 64 (44%) biopsies, 48 of 64 (75%; P < 0.001) perilesional swabs and 36 of 59 (61%; P = 0.04) buttock swabs. Forty-five different HPV types/putative types were detected: 15 were previously characterized HPV types, 17 were earlier described putative types and 13 were new putative types. In addition, six subtypes and four variants of HPV sequences were identified. HPV types within the B1 group (EV HPV types) were found in 26 of 64 (40%) lesions, 44 of 64 (69%) perilesional swabs and 35 of 59 (59%) buttock swabs. HPV 38 was detected in 23 of 59 (39%) patients, and was found in seven of 16 (43%) SKs, but was less common in SCCs [three of 23 (13%); P = 0.037] and BCCs [four of 25 (16%); P = 0.056]. Candidate HPV 92 was found in seven of 59 (12%) patients. CONCLUSIONS: A broad spectrum of HPV types, the majority from the B1 group, was found in skin of Australian patients with skin tumours. HPV 38 was found significantly more often in SK than in SCC. However, the role of cutaneous HPV infection in the pathogenesis of NMSC remains elusive. PMID- 12890197 TI - Melanoma-associated retinopathy: high frequency of subclinical findings in patients with melanoma. AB - BACKGROUND: Melanoma-associated retinopathy (MAR) is a paraneoplastic syndrome with symptoms of night blindness, light sensations, visual loss, defect in visual fields, and reduced b-waves in the electroretinogram. Patients with MAR often suffer from a sudden onset of ocular symptoms that are believed to result from antibody production against melanoma-associated antigens that cross-react with corresponding epitopes on retinal depolarizing bipolar cells. OBJECTIVES: To correlate the frequency of subclinical symptoms suggestive of MAR in melanoma patients to different stages of disease, patient age, type and thickness of the primary tumour, form of therapy, S-100 level and tumour burden. METHODS: We analysed 28 patients with melanoma in stages I-IV (according to the American Joint Committee on Cancer tumour classification) for the presence of subclinical MAR symptoms using scotopic electroretinography, static and kinetic perimetry and nyctometry. RESULTS: Seven patients had clinical signs and symptoms consistent with MAR, 18 had some indications, while the remaining three had none. We found no correlation between clinical symptoms and stage of disease, tumour burden or S 100 level, but findings suggestive of MAR were observed more frequently in advanced stages of disease. CONCLUSIONS: Subclinical retinal involvement characteristic of MAR appears to be more common than previously suspected in patients with cutaneous malignant melanoma. Our findings in this small cohort seem to indicate that the percentage of patients with symptoms suggestive of MAR is higher in advanced stages of disease. Further clinical studies are required to evaluate if the presence of subclinical symptoms suggestive of MAR is correlated with a worse prognosis and a shortened progression-free and overall survival. PMID- 12890198 TI - Long-term dermoscopic follow-up of melanocytic naevi: clinical outcome and patient compliance. AB - BACKGROUND: Digital dermoscopy for the follow-up of melanocytic naevi (MN) is becoming more common in dermatological private practice. OBJECTIVES: To evaluate the clinical outcome, including the patient's compliance, in a long-term follow up of single MN. METHODS: Criteria for the selection of MN for follow-up: clinically suspicious without dermoscopically atypical features, or typical for the patient. Clinical outcome measures: number of detected malignant melanomas (MM) and/or atypical MN; quantity, quality, and differences in morphological changes between 'low-risk' patients (no MM in history and < 50 MN) and 'high risk' patients (MM in history and/or > 50 MN). Compliance: the number of patients who joined a recommended follow-up scheme. RESULTS: No MM was found in 145 consecutive patients (mean age 28 years, 54% female) during a 4-year period (median follow-up per patient: 24 months; ranging 4-45; at least three visits). In five patients (3%), seven histologically proven atypical MN were shown on whole body examination at sites other than those documented. A total of 1968 images in 177 'low-risk' and 95 'high-risk' MN were analysed: 37% (n = 65) of 'low-risk' and 32% (n = 30) of 'high-risk' MN showed dermoscopic changes (difference not statistically significant), none were suspicious for MM. Compliance, evaluated within a separate database of 303 consecutive patients (mean age 32 years, 52% female) over a 6-month period, was only 46%, although recall letters were used. CONCLUSIONS: (i) In our setting of daily routines in dermatological private practices long-term follow-up of a single MN seems not to be helpful for the detection of MM. (ii) A whole body examination must be done at each visit. (iii) The clinician's experience of the type and number of possible morphological changes in MN is crucial in order to avoid unnecessary excisions. (iv) The patient's compliance might be an important problem. (v) Cost effectiveness has yet to be analysed. PMID- 12890199 TI - Frequency of sensitization to antimicrobials in patients with atopic eczema compared with nonatopic individuals: analysis of multicentre surveillance data, 1995-1999. AB - BACKGROUND: Atopic eczema, a multifactorial disease, is influenced by bacterial factors, particularly Staphylococcus aureus. Consequently, treatment includes therapy with antibiotics and nonantibiotic antimicrobials. OBJECTIVES: In our study the sensitizing potential of some topical antimicrobials was investigated by comparing proportions sensitized to various antibiotics and antiseptics between patients with current or past atopic eczema, and patients who had never had atopic eczema. METHODS: Data of all patients patch tested in the Departments of Dermatology participating in the Information Network of Departments of Dermatology (IVDK) between 1995 and 1999 were analysed. Patients with a current leg ulcer or stasis dermatitis that might cause a confounding effect were excluded. RESULTS: Controlling for the additional confounding effect of sex and particularly age, we did not find that patients with atopic eczema had a generally higher risk compared with nonatopic individuals. CONCLUSIONS: While the risk of sensitization should always be considered when applying topical therapy, our data-within the methodological limitations of this type of study-would not support a very restrictive use of these agents in the management of atopic eczema. PMID- 12890200 TI - Red ink tattoo reactions: successful treatment with the Q-switched 532 nm Nd:YAG laser. AB - BACKGROUND: In the South-west Thames region there were an unprecedented number of lichenoid tattoo reactions to red ink in patients who had visited a local tattoo parlour. The red ink was found to contain mercuric sulphide, a compound known to cause allergic reactions. Topical Dermovate (clobetasol propionate 0.05%, GlaxoWellcome) ointment alone had little impact. OBJECTIVES: To investigate whether the Q-switched 532 nm Nd:YAG laser could produce permanent flattening of the reaction. METHODS: This was an open nonrandomized clinical trial. Biopsies were taken from the lichenoid areas within the tattoos. Subjects were patch tested to 1% ammoniated mercury in petrolatum prior to treatment with the Q switched 532 nm Nd:YAG laser. Laser treatments were delivered at 6-weekly intervals by a single operator. Patients also applied topical Dermovate between treatments. Therapy was discontinued when the lesions flattened. Clinical photographs were assessed at baseline and prior to each laser treatment. RESULTS: Seven patients with Fitzpatrick skin types I-III were enrolled in the study (four females, three males, mean age 39 years). All patients completed the trial. Patch testing to mercury was universally negative at 48 and 96 h. Substantial flattening and depigmentation of the red ink within the tattoos was noted after six laser treatments. No adverse effects were recorded. CONCLUSIONS: The Q switched 532 nm Nd:YAG laser in combination with topical Dermovate ointment is a safe and effective method of treating red ink tattoo reactions. PMID- 12890201 TI - Trichilemmoma: an immunohistochemical study of cytokeratins. AB - BACKGROUND: The histogenesis of trichilemmoma remains unclear. OBJECTIVES: To clarify the histogenesis of trichilemmoma by evaluating its cytokeratin (CK) expression. METHODS: In three cases of trichilemmoma, CK expression was studied immunohistochemically using seven antikeratin antibodies against CK1, 10, 14-17 and 19, respectively. RESULTS: CK1 and CK10 were present in keratinizing ductal epithelium. CK14 was present in the whole layer. CK15 was present in suprabasal layers in two cases. CK16 was present in the suprabasal layer, but was absent in keratinizing ductal epithelium. CK17 was present in suprabasal layers and the sebaceous duct-like structure. CK19 was totally absent. CONCLUSIONS: These results showed that trichilemmoma may differentiate mainly towards two directions: infundibular keratinization and proliferation of the outer root sheath with undifferentiated and pluripotent characteristics. PMID- 12890202 TI - Comparison of the microvasculature of basal cell carcinoma and actinic keratosis using intravital microscopy and immunohistochemistry. AB - BACKGROUND: Angiogenesis is a prerequisite for growth of invasive tumours. We hypothesized that angiogenesis would be present in invasive basal cell carcinoma (BCC) but not in a noninvasive tumour such as actinic keratosis (AK). OBJECTIVES: To investigate both types of tumour for evidence of angiogenesis. METHODS: Patients with BCC or AK underwent intravital videocapillaroscopy. Three regions were examined: the tumour, perilesional skin and a control site. Microvessel width, area fraction and length density were determined from capillaroscopy images. Biopsies were stained for CD34 and a microvessel count was performed. RESULTS: Capillaroscopy demonstrated a grossly disorganized tumour microcirculation in BCC. Compared with control skin, microvessel width was increased 2.4-fold, area fraction was increased 4.9-fold and length density was increased 5.9-fold. In AK, microvessel width was increased 1.7-fold, area fraction 2.5-fold and length density 3.4-fold. Vessel width and area fraction were significantly greater in BCC than AK. Biopsies showed significant increases in microvessel length density for both BCC and AK compared with control skin, with BCC significantly greater than AK. CONCLUSIONS: Angiogenesis was demonstrated in BCC in humans in vivo, and to a lesser extent in AK. PMID- 12890203 TI - Legal claims in Scottish National Health Service Dermatology Departments 1989 2001. AB - BACKGROUND: An area of current National Health Service (NHS) interest is risk management as it is one of the requirements of clinical governance. If there are aspects of dermatological practice prone to mishap or dispute then a review of legal claims in dermatology may highlight these areas. This would then allow input into these specific areas of practice to try and minimize future risk. OBJECTIVES: To review all legal claims relating to NHS Dermatology in Scotland and characterize the main areas of risk. METHODS: Staff at the Central Legal Office in Edinburgh reviewed all legal claims in NHS Dermatology between 1989 and 2001. Details provided were general, and claimants, staff, hospitals and regions of Scotland could not be identified. RESULTS: Thirty claims were identified relating to five main areas of dermatological practice: (i) phototherapy (eight claims); (ii) therapeutics (eight); (iii) cryosurgery/cryotherapy (six); (iv) surgery (four); and (v) misdiagnosis (three) and one alleged neurological problem after fainting. As well as consultants, nurses featured in phototherapy claims and junior medical staff in cryosurgery claims. Twelve of the 30 (40%) claims have reached settlement; three claims are presently outstanding. CONCLUSIONS: Given the numbers of patients seen annually at Scottish NHS Hospital Dermatology units the absolute number of claims is low. Five main areas within dermatology were highlighted as at risk of litigation and this has implications for all U.K. dermatology departments with regard to funding, staff training and patient information and consent. PMID- 12890204 TI - Lifestyle differences in twin pairs discordant for basal cell carcinoma of the skin. AB - BACKGROUND: Basal cell carcinoma (BCC) of the skin is a multifactorial disease with a complex interplay of genetic, environmental and lifestyle factors, including patterns of solar exposure, with childhood exposure being of particular relevance. Case-control studies in disease-discordant twin pairs adjust for familial factors and improve the power of conventional case-control studies. OBJECTIVES: To investigate the roles of sociodemographic, dietary, medication, hormonal and other lifestyle factors in the aetiology of BCC. METHODS: The study base comprised all same-sex twins who had answered the baseline health questionnaire of the Finnish Adult Twin Cohort Study in 1975. Information on all histologically confirmed cases of BCC was retrieved from the nationwide cancer registry. The first twin to have BCC was defined as the case and his/her co-twin (alive at the time of diagnosis) as the control. Only pairs with the first BCC between 1976 and 1999 were included in the study (n = 333). Conditional logistic regression analyses were used to assess BCC risk by study factors. RESULTS: A significantly increased risk was associated with smoking status in females but not in males. The risk was higher in dizygotic than in monozygotic females, possibly indicating the presence of a gene-smoking interaction. Suggestive but statistically nonsignificant increases in risk were observed for higher education, urticaria and atopic eczema, use of nonsteroidal anti-inflammatory drugs and coffee. A suggestive risk decrease was observed for outdoor work. No clear effects were observed for marital status, body mass index, use of alcohol, tea, fruit and vegetables or oral contraceptives. CONCLUSIONS: The results suggest a role for smoking and possibly also for other lifestyle factors in the risk of contracting BCC. PMID- 12890205 TI - The effect of an iron chelating agent on protoporphyrin IX levels and phototoxicity in topical 5-aminolaevulinic acid photodynamic therapy. AB - BACKGROUND: In 5-aminolaevulinic acid (ALA)-photodynamic therapy (PDT), the prodrug ALA is endogenously converted to the active sensitizer protoporphyrin IX (PpIX), while further conversion of PpIX to haem requires iron. OBJECTIVES: To explore the potential of the iron chelator desferrioxamine (DFO) to enhance PpIX levels and phototoxicity in ALA-PDT. METHODS: A series of six doses of 2% ALA solution was iontophoresed into the healthy skin of each ventral forearm of 10 volunteers. One arm was pretreated with 20% DFO in aqueous cream, while the control arm received aqueous cream alone, for 16 h. At 5 h following iontophoresis, skin-surface PpIX fluorescence was measured, following which the forearms were simultaneously irradiated with 100 J cm-2 broadband red light. The phototoxic reaction was assessed at 24 h postirradiation as the minimal phototoxic dose (MPD) and with quantification of erythema. Next, eight patients with two superficial basal cell carcinomas or two plaques of Bowen's disease of similar appearance received 20% ALA topically to one lesion and 20% ALA with 20% DFO to the other, for 3 h. Skin-surface PpIX fluorescence was measured at 5 h, following which lesions were irradiated with 100 J cm-2 broadband red light. RESULTS: In healthy skin, PpIX fluorescence increased with increasing ALA dose at DFO-treated and untreated sites (P < 0.0005); PpIX fluorescence peak values were consistently higher in DFO-treated compared with control sites (P < 0.02). Erythema also correlated with ALA dose (P < 0.0005), but a significant difference between active and control sites occurred only at low ALA dose (P < 0.05). The median MPD appeared lower at the DFO-treated sites, at 6 mC vs. 12 mC (P = 0.06). In contrast, in lesional skin there was no consistent difference in PpIX fluorescence levels between those treated with and without DFO. CONCLUSIONS: While iron chelation augmented ALA-PDT phototoxicity in normal skin, this occurred only at low ALA dose. Addition of DFO does not appear to confer additional benefit in ALA-PDT of nonmelanoma skin cancers. PMID- 12890206 TI - Photosensitivity in patients with lupus erythematosus: a clinical and photobiological study of 100 patients using a prolonged phototest protocol. AB - BACKGROUND: There is a clear relationship between ultraviolet (UV) radiation (UVR) and the clinical manifestations of patients with lupus erythematosus (LE). Cutaneous lesions are induced or exacerbated by exposure to UVR. Of patients with LE, 24-83% are reported to be photosensitive to UVR. LE tumidus appears to be the most photosensitive subtype of LE, followed by subacute cutaneous LE (SCLE). In general, the history of patients with LE correlates poorly with the presence or absence of photosensitivity, due to a delayed time interval between UV exposure and exacerbation of skin lesions. Phototesting using artificial UVR and visible light is a reliable way of diagnosing photosensitivity. OBJECTIVES: To investigate the photoreactivity of patients with various subtypes of LE using an individualized phototest protocol. The results of phototests were correlated with the history of photosensitivity, the subtype of LE, the presence of autoantibodies and the use of anti-inflammatory medication by these patients. METHODS: Phototesting with UVA, UVB and visible light was performed in 100 patients with LE. The diagnosis of LE was established both on clinical examination and skin histology. Serological studies were also performed in all patients. The phototests were performed on large skin areas of the forearm or trunk; the first dose was twice the minimal erythema dose and the dosage was increased according to the individual reactions of the patients at the test sites. Follow-up of skin reactions at the test sites was performed for up to 2 months. Histological examination of the photoprovoked skin lesions was carried out in 57 patients. RESULTS: Of the 100 patients included (81 women and 19 men; mean age 41 years, range 17-79), 46 had chronic discoid LE, 30 SCLE and 24 systemic LE. An abnormal reaction to UVR and visible light was found in 93% of our patients with LE. No clinical or histological evidence at the phototest sites of polymorphic light eruption was found. There was no correlation between photosensitivity and LE subtype, presence of autoantibodies or medical history. Concomitant use of anti-inflammatory medication seemed to exert only minimal influence on the results of phototesting. CONCLUSIONS: When using an extended phototesting protocol, almost all patients with LE in this study showed clinical and histological evidence of aberrant photosensitivity. Therefore, patients with LE should receive thorough advice and instruction on photoprotective measures, regardless of their history, LE subtype or presence of autoantibodies. PMID- 12890208 TI - Combination TL01 ultraviolet B phototherapy and topical calcipotriol for psoriasis: a prospective randomized placebo-controlled clinical trial. AB - BACKGROUND: Previous studies have demonstrated the ultraviolet (UV)-sparing effect of combining topical calcipotriol with broadband UVB in the treatment of psoriasis. OBJECTIVES: To determine if the combination of narrowband TL01 UVB phototherapy and topical calcipotriol produces the same UVB-sparing effect. METHODS: This was a randomized, placebo-controlled, blinded clinical trial. Fifty psoriasis patients were recruited, 25 of whom were randomized into the active group who received TL01 phototherapy together with twice-daily application of calcipotriol cream 50 microg g(-1). The control group received TL01 phototherapy and twice-daily application of a topical emollient as placebo. TL01 phototherapy was given three times per week starting at 70% minimal erythema dose with 20% increments as tolerated for up to approximately 20 sessions. Patients were assessed using the Psoriasis Area and Severity Index (PASI) and Psoriasis Disability Index (PDI). They were evaluated at treatment sessions 8, 14 and 20, and followed up at 5 and 10 weeks post-treatment. Statistical analysis was performed using a two-tailed t-test. RESULTS: There were no significant differences in demographic characteristics and baseline PASI and PDI scores between the two groups. The mean PASI score declined significantly (P < 0.01) for both groups after treatment. The difference in mean PASI score reduction from baseline between the two groups was only significant during the first eight sessions, with a net reduction of 3.6 (95% confidence interval 1.0-6.2, P = 0.008) in the active group relative to the control group. The mean PDI score declined significantly (P < 0.05) for both groups, but there was no statistical difference in mean PDI score reduction between the two groups (P = 0.8) at the end of treatment. The mean cumulative UVB dose for the active group was significantly lower (P < 0.02) at 16 204 mJ cm-2 compared with 21 082 mJ cm-2 for the control group. CONCLUSIONS: We conclude that combining TL01 phototherapy with topical calcipotriol cream has a UVB-sparing effect. PMID- 12890207 TI - An evaluation of the usefulness of mycophenolate mofetil in pemphigus. AB - BACKGROUND: Pemphigus is a group of autoimmune blistering diseases of the skin and/or mucous membranes requiring management with immunosuppressive therapy. The optimal therapeutic regimen would rapidly induce remission and maintain effectiveness with minimal adverse effects in the long term. OBJECTIVES: The present study describes our experience of the addition of mycophenolate mofetil (MMF) to prednisolone in the management of severe, refractory pemphigus. METHODS: Patients with active, refractory pemphigus were treated with MMF. Our series included 12 cases of pemphigus vulgaris, four cases of pemphigus foliaceous and one case of paraneoplastic pemphigus. All patients were monitored to assess disease control and mycophenolate toxicity. RESULTS: Of the 17 cases, MMF has been of benefit to 12. MMF was well tolerated and there were no treatment withdrawals because of safety concerns. CONCLUSIONS: We found that MMF permitted a reduction in prednisolone dosage without disease relapse. PMID- 12890209 TI - A randomized trial of amorolfine 5% solution nail lacquer in association with itraconazole pulse therapy compared with itraconazole alone in the treatment of Candida fingernail onychomycosis. AB - BACKGROUND: Treatment failures and relapses are not uncommon in onychomycosis. Therefore, it is worthwhile to consider the combination of systemic and topical antifungals to improve the cure rates further and to reduce the duration of systemic treatment. OBJECTIVES: To evaluate and compare itraconazole pulse therapy combined with amorolfine with itraconazole alone in the treatment of Candida fingernail onychomycosis. METHODS: Ninety patients with moderate to severe Candida fingernail onychomycosis were randomized into two treatment groups of 45 subjects each. Group 1 received itraconazole pulse therapy for 2 months and applied amorolfine 5% solution nail lacquer for 6 months, while group 2 received monotherapy with three pulses of itraconazole. The primary efficacy criterion was the result of mycological examination at 3 months. The secondary criterion was the combined mycological and clinical response at 9 months. A pharmacoeconomic analysis was also performed to compare the cost-effectiveness of combined therapy vs. monotherapy. RESULTS: Eighty-five patients were analysed (73 women and 12 men, mean +/- SD age 44.2 +/- 12.9 years). Patients had a mean +/- SD of 3.64 +/- 2.0 nails involved and 228.6 +/- 148.0 mm2 of their nail surface diseased. The mean duration of onychomycosis was 11 months. Paronychial involvement was evident in 71 patients. C. albicans was isolated in 85 cases, C. parapsilosis in three and other Candida species in two cases. Side-effects were uncommon and in only one case led to withdrawal. At the 3-month visit, mycological cure was seen in 32 (74%) of 43 patients in group 1 and in 25 (60%) of 42 patients in group 2. At the 9-month visit, a global cure was seen in 40 patients (93%) in group 1 and in 34 patients (81%) in group 2. Statistical analysis showed no statistically significant difference (P > 0.1) between the two treatment groups. The cost per cure ratio was 1.63 and 1.70euro for groups 1 and 2, respectively. CONCLUSIONS: The combination of amorolfine and oral itraconazole, which interfere with different steps of ergosterol synthesis, exhibited substantial synergy. Compared with oral itraconazole alone, the combination achieved greater mycological cure and increased total cure rate. However, no statistically significant difference was documented for this number of observations. Combination treatment with amorolfine and two pulses of itraconazole is at least as safe and effective as three pulses of itraconazole, with a lower cost per patient. In our opinion, the addition of amorolfine to oral itraconazole pulse therapy is of value in the treatment of moderate to severe Candida fingernail onychomycosis. PMID- 12890210 TI - Seasonality trends of scabies in a young adult population: a 20-year follow-up. AB - BACKGROUND: Scabies is not a notifiable disease in most countries. Therefore, its seasonality trends are usually estimated and may be inaccurate. OBJECTIVES: To determine the seasonality of scabies over the long term in a large military population, including all of the Israel Defence Forces personnel. METHODS: For the last two decades, the Israel Defence Forces have mandated the routine reportage of every case of scabies infestation in soldiers to the Epidemiology Department of the Army Health Branch. This database was used in the present study to analyse the incidence of scabies by season. Scabies was defined according to the report by a military physician based on the typical clinical presentation, relevant epidemiological data and, in most cases, microscopic visualization of the mite. RESULTS: The person-time incidence of scabies was higher in winter than summer (P < 0.001). The overall risk ratio was 1.31 (95% confidence interval 1.28 1.33). Sensitivity analysis indicated that no single year was an important source of disease heterogeneity. CONCLUSIONS: Scabies in a young adult population is more frequent in the cooler months of the year. PMID- 12890211 TI - Circulating natural killer cells in psoriasis. AB - BACKGROUND: Psoriasis is an immunologically mediated, probably autoimmune, disease in which T-helper type 1 cytokines play an important role. Established autoimmune diseases, with similar mechanistic characteristics to psoriasis, include multiple sclerosis, rheumatoid arthritis, type 1 diabetes mellitus and systemic lupus erythematosus. Natural killer (NK) and natural killer-T (NK-T) cells are considered key to the pathogenesis of these conditions, which are characterized by reduced numbers of NK cells in peripheral blood. NK and NK-T cells have been implicated in the pathogenesis of psoriasis and are present in plaques of psoriasis. OBJECTIVES: To investigate whether levels of NK and NK-T cells are reduced in the peripheral blood of patients with psoriasis. METHODS: Fourteen patients with untreated psoriasis, mean +/- SD age 46 +/- 13 years, and 13 healthy volunteers, mean +/- SD age 34 +/- 9 years, were venesected and peripheral blood mononuclear cells isolated, labelled with a panel of antibodies to T-cells and NK cells including CD3, CD56, CD57, CD16, CD94, CD158a, CD69 and cutaneous lymphocyte-associated antigen (CLA) and analysed using triple-colour flow cytometry. RESULTS: There were significantly fewer cells expressing the NK cell markers CD16 (P < 0.001), CD56 (P < 0.003), CD94 (P < 0.001) and CD158a (P < 0.02) in patients with psoriasis compared with normal controls. However, circulating numbers of NK-T cells (CD3+ CD56+ CD57+), T-cells (CD3+), activated lymphocytes (CD69+) or CLA+ cells were not significantly different between patients with psoriasis and controls. CONCLUSIONS: Circulating NK cells are reduced in psoriasis. This finding is similar to those in established autoimmune diseases such as rheumatoid arthritis. This observation provides some evidence that psoriasis may be an autoimmune disease in which NK cells play a role. PMID- 12890212 TI - Carbamazepine-induced hypersensitivity syndrome associated with transient hypogammaglobulinaemia and reactivation of human herpesvirus 6 infection demonstrated by real-time quantitative polymerase chain reaction. AB - Drug-induced hypersensitivity syndrome (HS) is a rare but severe disease with multiorgan failure. Many different precipitating factors have been reported, but the pathophysiology of HS remains unknown. However, the association of the human herpesvirus (HHV) family, particularly of HHV-6, has recently been reported in patients with HS. We report a 14-year-old boy who was diagnosed as having carbamazepine-induced HS based on the clinical course, laboratory data and results of drug-induced lymphocyte stimulation tests. In addition, the reactivation of HHV-6 was demonstrated by real-time quantitative polymerase chain reaction and by significantly increased levels of the specific antibody in his paired sera. Furthermore, transient hypogammaglobulinaemia was detected in the early stage of the disease. In addition, serum levels of interferon-gamma, interleukin (IL)-6, IL-5 and eosinophil cationic protein, which were increased on admission, decreased dramatically after steroid therapy. This is the first report of carbamazepine-induced HS associated with reactivation of HHV-6, transient hypogammaglobulinaemia, increased serum levels of inflammatory cytokines and activated eosinophils. This case might contribute to the understanding of the pathophysiology of HS. PMID- 12890213 TI - CD30+ T-cell lymphoma in a patient with psoriasis treated with ciclosporin and infliximab. AB - There is a known relationship between the use of immunosuppressive therapies and the development of lymphoproliferative malignancies. These lymphomas are mainly B cell nonHodgkin's lymphomas associated with Epstein-Barr virus. Most cases concern classical immunosuppressive treatments including ciclosporin and methotrexate. A relationship between the new antitumour necrosis factor (TNF) alpha agents and lymphoproliferative malignancies is debated. Patients with psoriasis on immunosuppressive therapies, mainly ciclosporin, are considered to have a low risk of developing lymphoid proliferation. We report a patient with erythrodermic psoriasis treated with ciclosporin and infliximab who developed a CD30+ T-cell lymphoma. This lymphoma regressed after stopping these treatments. In this case, the anti-TNF-alpha agent may have played a role in association with ciclosporin in the development of the lymphoproliferative disorder. Whereas the combination of anti-TNF-alpha therapies with methotrexate has been well studied, their combination with ciclosporin has been evaluated only in a few patients. Psoriatic patients who may require anti-TNF-alpha treatment have often been or will be treated with ciclosporin. The combination of ciclosporin and anti-TNF alpha warrants further investigation. PMID- 12890214 TI - An autosomal recessive exfoliative ichthyosis with linkage to chromosome 12q13. AB - A new variant of congenital exfoliative ichthyosis in two related Bedouin families is reported. The ichthyosis appeared shortly after birth as a fine peeling of nonerythematous skin on the palms and soles. The prominent well demarcated areas of denuded skin in moist and traumatized regions resembled the 'mauserung' phenomenon of ichthyosis bullosa of Siemens (IBS). Unlike in IBS, epidermolysis is absent on histological examination. Electron microscopy revealed a prominent intercellular oedema and numerous aggregates of keratin filaments in basal keratinocytes. Abnormal keratin (K) 1 expression was seen in the affected epidermis; however, all other keratins, including K2e, had a distribution comparable to that seen in normal controls. A maximum two-point LOD score of 2.53 and multipoint LOD score of 3.76 were obtained for marker D12S390, suggesting linkage to the type II keratin cluster on chromosome 12q13. Sequencing of both the K1 gene, the promotor and the 3' calcium regulatory region did not reveal a mutation. K2e and K5 genes, as well as the genes harboured within the minimal region, such as retinoic acid receptor gamma, sterol O-acyltransferase 2, integrin beta7 and insulin-like growth factor binding protein-6, were also excluded. This combination of clinical, histological, ultrastructural and genetic features has not been previously reported in other congenital exfoliative ichthyoses. We therefore suggest that it represents a new form of exfoliative ichthyosis. PMID- 12890215 TI - Successful treatment with infliximab and methotrexate of pyostomatitis vegetans associated with Crohn's disease. AB - A 35-year-old woman with severe fistulizing Crohn's disease presented with pyostomatitis vegetans affecting both the mouth and the vulva. The coalescing pustules transformed within several days into vegetating lesions on areas of inflammation. Microbial assessments revealed no pathogenic agent. Histology showed neutrophilic microabscesses, but no granulomas. Three injections of infliximab and maintenance therapy with methotrexate resulted in rapid and complete regression of both the pyostomatitis vegetans and the Crohn's disease. Infliximab and methotrexate may be a promising treatment for the rare cases of pyostomatitis vegetans associated with Crohn's disease. PMID- 12890216 TI - A Japanese case of segmental Darier's disease caused by mosaicism for the ATP2A2 mutation. AB - Darier's disease is an autosomal dominant skin disorder that is characterized by multiple keratotic papules, focal loss of adhesion and abnormal keratinization. Mutations in the ATP2A2 gene encoding sarco/endoplasmic reticulum calcium pumping ATPase type 2 have been identified as the molecular basis of Darier's disease. Segmental Darier's disease is a rare type of Darier's disease in which there is characteristic localization of the keratotic papules in a linear pattern following Blaschko's lines. In this study we examined ATP2A2 mutations in a Japanese patient with segmental Darier's disease. The samples from affected skin, unaffected skin and peripheral leucocytes were subjected to polymerase chain reaction (PCR). Direct sequencing of the PCR products was performed. Sequence analysis revealed that the patient had 160A-->G substitution mutation which predicts I54V. This novel mutation was present in the affected skin, but not in the unaffected skin or peripheral leucocytes. This is the first report of segmental Darier's disease caused by mosaicism for an ATP2A2 mutation in Japan. PMID- 12890217 TI - Atypical exanthem: polymerase chain reaction spots the cause. PMID- 12890218 TI - An unusual presentation of congenital self-healing reticulohistiocytosis. PMID- 12890219 TI - Bexarotene reverses alopecia in cutaneous T-cell lymphoma. PMID- 12890220 TI - Immunohistological characterization of a Japanese case of pityriasis rotunda. PMID- 12890221 TI - Ciclosporin A-induced sebaceous gland hyperplasia. PMID- 12890222 TI - Disseminated cutaneous Mycobacterium haemophilum infection with severe hypercalcaemia in a failed renal transplant recipient. PMID- 12890223 TI - Juvenile pityriasis rubra pilaris: successful treatment with ciclosporin. PMID- 12890224 TI - Neurogenic pruritus: a case of pruritus induced by transverse myelitis. PMID- 12890225 TI - Early apoptosis plays an important role in the healing mechanism of cutaneous basal cell carcinomas after photodynamic therapy. PMID- 12890226 TI - Dead Sea extract sold under-the-counter. PMID- 12890227 TI - Tazarotene 0.1% gel for psoriasis of the fingernails and toenails: an open, prospective study. PMID- 12890228 TI - Stigmatization and psoriasis. PMID- 12890229 TI - Suggestions for uniform outcome variables when reporting treatment effects in hidradenitis suppurativa. PMID- 12890230 TI - High-level ultraviolet A photoprotection is needed to prevent doxycycline phototoxicity: lessons learned in East Timor. PMID- 12890231 TI - Cold urticaria responding to systemic ciclosporin. PMID- 12890232 TI - Severe bullous drug eruption due to sibutramine (Reductil). PMID- 12890233 TI - Unusual, rapidly growing ulcerative genital mass due to herpes simplex virus in a human immunodeficiency virus-infected woman. PMID- 12890234 TI - Share of research output in dermatology: a quantitative ranking. PMID- 12890235 TI - Efficacy of fumaric acid ester monotherapy in psoriasis pustulosa palmoplantaris. PMID- 12890236 TI - Granulomatous blepharitis successfully treated with tranilast. PMID- 12890237 TI - Topical treatment of Netherton's syndrome with tacrolimus ointment without significant systemic absorption. PMID- 12890238 TI - Image-guided spinal injection for diagnosis and therapy. AB - Over the past decade, major advances have been made in our ability to identify a putative cause of patients' spinal pain using image-guided injection techniques. These techniques might also provide a means of temporarily relieving patients' pain and facilitating increased mobility. This article aims to review the rationale for and the techniques of the more commonly used spinal injection techniques. PMID- 12890239 TI - Retinoblastoma: review of 30 years' experience with external beam radiotherapy. AB - A review of the experience at the Peter MacCallum Cancer Centre (Peter Mac), Melbourne, Australia in treating retinoblastoma with external beam radiotherapy was conducted. Outcomes of particular interest were tumour control, vision preservation and treatment late effects. The review was restricted to patients that had intact eyes treated at Peter Mac from 1965 until 1997 with at least 2 years of follow up. Histories were reviewed regarding patient and tumour characteristics and treatment details. Thirty-five patients were identified in whom 47 eyes were treated. Of the tumours, 47% were Reese-Ellsworth stage IV or V and the majority of others were at high risk for vision loss because of tumour location. The radiation treatment technique became increasingly sophisticated during the study period. Radiation dose and fraction size have similarly evolved but most patients received 30-50 Gy. Since 1989, a highly accurate contact lens immobilization technique has been used to deliver 40 Gy in 20 fractions. Thirteen eyes required additional local therapy. Of the treated eyes, 34 (72%) remain intact and 74% of these have useful vision. One patient died from retinoblastoma and three from second malignant neoplasms. With modern radiotherapy, late toxicities other than growth arrest and non-progressive cataract did not occur during the study period. Tumour control was high and a very acceptable rate of organ and vision preservation was achieved in a relatively high-risk population. Modern radiotherapy continues to develop in an attempt to improve treatment accuracy and minimize late radiation toxicity. PMID- 12890240 TI - Emergency embolization of spontaneous ruptured hepatocellular carcinoma: correlation between survival and Child-Pugh classification. AB - From February 1994 to April 2000, 29 emergency gelfoam embolizations for spontaneous ruptured hepatocellular carcinoma (HCC) performed in 28 patients were retrospectively reviewed. There were 11 patients in Child's A, 11 in Child's B and six in Child's C classification of cirrhosis. The duration of the procedure, artery embolized and complications were reviewed, and the Child-Pugh classification of each patient was correlated with their mean survival period. Embolization was done in 12 right hepatic arteries, two left hepatic arteries and 15 proper hepatic arteries. In one patient, the left hepatic artery was embolized initially but the proper hepatic artery was also embolized because another subcapsular liver tumour was found after reviewing the preangiogram CT scan. The entire procedure took 40-170 min (mean = 86 min) with no periprocedural complication. Following embolization, the mean survival period for Child's A class was 218.3 days, Child's B class was 83.4 days and Child's C class was 11.0 days. Transcatheter embolization is an effective treatment to arrest bleeding in spontaneous ruptured HCC. Patients with Child's A class cirrhosis have the longest survival. Selective embolization of either the right or the left hepatic artery alone carries the potential risk of missing multifocal HCC that might not be easily appreciated during angiography. PMID- 12890241 TI - Precision of bone densitometry measurements: when is change true change and does it vary across bone density values? AB - The precision error of the bone densitometer is used to interpret significant change in bone mineral density (BMD) in serial studies. The precision error can be expressed as standard deviation (SD) or coefficient of variation (CV). The aims of this study are to determine the precision error over a range of BMD values and to demonstrate the application of the precision error in clinical practice. A bone phantom was used consisting of a perspex block with eight compartments containing varying amounts of hydroxyapatite powder to simulate a range of bone densities. The block was scanned 21 times and manual regions placed over each compartment to measure the BMD in each compartment. There were no significant differences in the variances or SD for all eight compartments, that is, over the range of BMD normally encountered in clinical practice. However, the calculated CV show a progressive fall in values as the BMD rises. Therefore, the SD should be used to calculate significant BMD change. In a practise with quality control procedures in place to detect calibration drift and with appropriately trained personnel, a change of approximately 0.05 g/cm2 is generally regarded as being a significant change at a 95% confidence level. PMID- 12890242 TI - Development of an evidence-based guideline for imaging in cervical spine trauma. AB - Cervical spine trauma is a common reason for presentation to an emergency department. However, less than 5% of patients who have suffered possible neck injury actually have an injury requiring medical treatment. Nevertheless, the consequences, both for the patient and the doctor, of a missed injury are well recognized by emergency department medical staff. This results in the vast majority of these patients receiving some form of diagnostic imaging. We describe the development of an evidence-based imaging guideline for use in the patient who has suffered cervical spine trauma. The guideline aims to help clinicians determine, at the bedside, when it is appropriate to use imaging and which imaging modality should be used first. Correct utilization of the guideline should lead to a reduction in the number of imaging tests required to reach a diagnosis without adverse patient outcomes. PMID- 12890243 TI - Hydrodilatation in the management of shoulder capsulitis. AB - The aim of this study was to research the benefit of hydraulic arthrographic capsular distension (hydrodilatation) in the management of adhesive capsulitis of the shoulder. One hundred and nine shoulders with primary adhesive capsulitis were treated with hydrodilatation. Prior to the procedure, 93 shoulders were painful. Two months following the procedure, 31 continued to have some pain. In the 109 shoulders, the measured range of passive glenohumeral movement improved by approximately 30 degrees in all directions. The procedure was of similar benefit if carried out early or late in the disease process. The absolute improvement in movement range was similar in severe and mild cases. The severe cases in the long term, although improved, still had more restriction in movement and tended to have more pain than the other cases. There was considerable improvement in all the non-diabetic patients. The patients with diabetes responded less well in the long term to hydrodilatation and had an increased requirement for arthroscopic surgery. Effective treatment of adhesive capsulitis can be achieved in the majority of cases with an immediate hydrodilatation of the shoulder. Technically, it is important to achieve maximum distension, preferably with capsular rupture, and to utilize cortisone in the fluid injected. PMID- 12890244 TI - Triangular cord sign and ultrasound features of the gall bladder in infants with biliary atresia. AB - The aim of this study was to reassess the accuracy of the triangular cord sign, the triangular cord sign coupled with abnormal gall-bladder length, and an irregular gall-bladder wall in the diagnosis of biliary atresia. The ultrasonograms of 46 infants with cholestatic jaundice were reviewed for the triangular cord sign, gall-bladder length and gall-bladder wall without knowledge of the clinical data. Of the 23 infants with biliary atresia, 22 had the triangular cord sign whereas 17 infants with other causes of cholestatic jaundice did not have the triangular cord sign. The sensitivity, specificity, accuracy and positive predictive value of the triangular cord sign in the diagnosis of biliary atresia were 95.7, 73.9, 84.8 and 78.6%, respectively. The sensitivity, specificity, accuracy and positive predictive value of the triangular cord sign coupled with abnormal gall-bladder length in the diagnosis of biliary atresia were all 95.7%. Gall-bladder wall irregularity was seen in seven of 14 infants (50%) with biliary atresia whose gall bladders contained bile on ultrasound and in two of 22 infants (9.1%) without biliary atresia whose gall bladders contained bile on ultrasound. At the medical centre where this study was performed and where infants present with cholestatic jaundice at an advanced stage, the ultrasonographic triangular cord sign coupled with abnormal gall-bladder length is more reliable than the ultrasonographic triangular cord sign alone or gall bladder wall irregularity in the diagnosis of biliary atresia. PMID- 12890245 TI - Radiologists' detection of mammographic abnormalities with and without a computer aided detection system. AB - The aim of this study was to to evaluate the role of a computer-aided program (CAD) in assisting detection of mammographic lesions by radiologists not specifically trained in mammography and its potential utility in breast screening. Mammograms were evaluated by radiologists not specifically trained in mammography first without, and then with, the CAD. Finally, the mammograms were evaluated by experienced mammographers who formed the reference standard. Two hundred and ninety four breasts were studied. In 257 breasts (87.4%), the CAD system did not help with the detection of abnormalities. It assisted radiologists in detecting abnormalities in 21 breasts (7.1%) with potential for detecting abnormalities in a further 13 breasts (4.4%). Only in three cases (1%) did it cause confusion in interpretation. There is overall increased sensitivity in detecting mammographic abnormalities with the aid of the CAD system from 74.4 to 87.2%, which is statistically significant. However, it failed to detect suspicious abnormalities in 71 breasts (24.1%). The CAD system improved detection of suspicious mammographic abnormalities by radiologists who are not specifically trained in mammography. However, there is also a substantial failure to detect suspicious mammographic features that cautions against over-reliance on the system, emphasizing its role as a second reader at best. PMID- 12890246 TI - Stereotactic breast biopsy: an audit of 18 months at BreastScreen Auckland. AB - Stereotactic biopsy has become a widely used technique for marnmographically detected lesions that are clinically and sonographically occult. Vacuum-assisted (mammotome) biopsy on a dedicated prone stereotactic unit has further increased ease of biopsy and utility of the technique. Results of an 18-month audit in the national breast screening programme (BreastScreen Auckland and North) in the Auckland region demonstrates results comparable with those achieved elsewhere. Of a total of 399 stereotactic biopsies, 23 were excluded as they were 14-G core biopsies rather than 11-G mammotome biopsies. Of 376 mammotome biopsies, 10 (2%) failed, 266 (70.7%) were benign, 39 (10.3%) were atypical ductal hyperplasia (ADH) and 70 (18.6%) were malignant. Of these, 14.6% were ductal carcinoma in situ (DCIS) and 4% were invasive carcinomas. All cases diagnosed as ADH underwent formal excision biopsy. Of the 39 cases, 33 demonstrated benign disease or residual ADH only on excision (84.6%) and six (15.4%) patients were upgraded: five to DCIS and one to invasive carcinoma. The significant complication rate was 0.5%. Indications for biopsy were calcification in 89% of cases, mass lesions in 10.6% of cases and architectural distortion in 0.2% of cases. The failure rate of 2% compares with hookwire biopsy series. Practical issues and protocols have been presented. PMID- 12890247 TI - Are radiologists meeting the needs of Australian medical oncologists? Results of a national survey. AB - Although radiological evaluation plays an integral role in the management of oncology patients, little is known about which elements of such evaluation are most important or about how satisfied clinicians are with the quality of radiology services in cancer patients. We have developed a 36-item anonymous survey evaluating availability of and satisfaction with radiology services to medical oncologists. The survey was distributed to members of the Medical Oncology Group of Australia resident in Australia. We received 124 responses (51%). Most respondents (76.9%) were satisfied with their radiology services. Satisfaction correlated closely with the availability of standardized reporting and promptness of reporting (P < 0.001). Oncologists in private practise were more likely to receive prompt reports (P < 0.0001). Oncologists in public practice were more concerned about availability of CT scanning (P = 0.02). Services that were reported as less frequently available, despite being ranked as very important, included measurements of key lesions and comparison to previous studies. Standardized reporting was considered helpful by 91% of respondents. Only 32.5% of respondents reported receiving such a summary in more than 50% of cases and 21% never received such a summary. Our findings highlight the need for closer collaboration between oncologists and radiologists in order to address specific needs and develop optimal patterns of practise. Consideration should be given to standardization of reporting of radiology studies in oncology. PMID- 12890248 TI - Utility-adjusted analysis of the cost of palliative radiotherapy for bone metastases. AB - Palliative radiotherapy is effective in the treatment of bone metastases but is under-utilized, possibly because it is perceived to be expensive. We performed a cost-utility analysis of palliative radiotherapy for bone metastases, evaluating both the actual cost of radiotherapy as well as its impact on quality of life by adjusting for the variation in response to treatment. Hospital records between July 1991 and July 1996 were reviewed to ascertain the number of patients treated with palliative radiotherapy for bone metastases, the average number of fields of radiation delivered to each patient and the average duration of survival. Partial and complete response rates to palliative radiotherapy were obtained from a review of all published randomized controlled trials of radiation treatment of bone metastases. Utility values were assigned to the response rates, and an overall adjusted response rate to radiotherapy was derived. The cost of delivering a field of radiation was calculated. The total cost was divided by the total number of response months to give a utility-adjusted cost per month of palliative radiotherapy. The utility-adjusted cost per month of palliative radiotherapy of bone metastases was found to be AUS dollars 100 per month or AUS dollars 1200 per utility-adjusted life-year. This study demonstrates that, contrary to popular perception, palliative radiotherapy is a cost-effective treatment modality for bone metastases. PMID- 12890249 TI - Early cervical cancer treated with definitive or adjuvant radiotherapy: improved survival with adjuvant radiotherapy attributable to patient selection. AB - The optimum management of patients with early cervical cancer (Federation Internationale de Gynecologie Stages IB and IIA) remains controversial. We reviewed our radiotherapy practise and compared patients with early stage cervical cancer who had surgery and adjuvant radiotherapy (ART) to those that had definitive radiotherapy (DRT). One hundred and twenty-seven patients were identified, 81 of these underwent ART and 46 underwent DRT. Patients who underwent DRT were significantly older and of poorer performance status than those who underwent ART. The 5-year relapse-free survival in the ART and DRT groups were 79 and 72%, respectively (P = 0.70). The corresponding 5-year overall survival figures were 86 and 58% (P = 0.006). The difference was due to increased deaths from other causes in the DRT arm, 37 versus 7% (P = 0.0007.) The poorer overall survival of DRT patients was due to patient selection. PMID- 12890250 TI - A prospective trial of short-fractionation radiotherapy for the palliation of liver metastases. AB - The purpose of this study was to prospectively examine the effectiveness and tolerability of a simple radiotherapy technique for the palliation of symptomatic liver metastases. Twenty-eight patients with symptomatic liver metastases were enrolled from seven centres, and received targeted (partial or whole) liver irradiation consisting of 10 Gy in two fractions over 2 days. Symptoms at baseline were hepatic pain (27 patients), abdominal distension (19), night sweats (12), nausea (18) and vomiting (eight). Twenty-two patients (76%) had failed previous treatment with chemotherapy, hormonal therapy and/or high-dose steroids. Symptoms and potential toxicities were prospectively assessed at the time of treatment, then 2, 6 and 10 weeks later. Individual symptom response rates were 53-66% at 2 weeks. Partial or complete global symptomatic responses were noted in 15 patients (54%) overall. The treatment was well tolerated with two patients (7%) experiencing grade 3 toxicity (one vomiting and one diarrhoea); however, four patients reported temporary worsening of pain shortly after treatment. This simple and well-tolerated treatment achieves useful palliation. PMID- 12890251 TI - Multicentre dosimetric comparison of photon-junctioning techniques in head and neck radiotherapy. AB - Because many head and neck radiotherapy treatment techniques rely on a junction between X-ray fields, it was the aim of the present study to investigate the use of different junctioning techniques and the affect on the dose across the junction. Techniques in use at nine radiotherapy centres in Australia were investigated using thermoluminescence dosimetry (TLD). The techniques could broadly be divided into two groups: (i) use of the light field to match the fields after moving the patient; and (ii) use of asymmetric collimation to create a single isocentre located in the junction. The mean dose at the junction and its reproducibility was studied in five consecutive treatments in each centre using 25 TLD chips placed throughout the junction in an anthropomorphic phantom. There was a tendency for the mono-isocentric technique to deliver a lower, more accurate mean dose at the junction (Group I: 1.22 Gy (n = 8) vs Group II: 0.96 Gy (n = 5) for 1 Gy planned, some centres contributed to both technique) with greater reproducibility (Group I: 9.6%, Group II: 5.1% of the mean dose). We conclude that a mono-isocentric treatment technique has the potential to deliver a more accurate and reproducible dose distribution at the field junction of photon beams in head and neck treatment. PMID- 12890252 TI - Neurosarcoidosis of the conus medullaris and cauda equina. AB - An uncommon case of neurosarcoidosis of the conus medullaris and cauda equina is described in a 24-year-old man with a 4-year history of urological symptoms and rash. Chest X-ray and biopsy confirmed sarcoidosis and MRI diagnosed conus medullaris involvement. PMID- 12890253 TI - Spoke-wheel pattern in renal oncocytoma seen on double-phase helical CT. AB - Renal oncocytomas are benign, solid tumours of the kidney. An angiographic spoke wheel pattern is known to be associated with oncocytomas, although it is not pathognomonic. On review of the literature, we found two reports of sonographic spoke-wheel appearance in oncocytomas. These were sufficiently characteristic to enable a confident preoperative diagnosis of oncocytoma. We present a case of a surgically proven oncocytoma with a distinct helical CT appearance, -commensurate with the angiographic and sonographic spoke-wheel appearance from which the diagnosis was suspected preoperatively. PMID- 12890254 TI - Uterine arteriovenous malformation: a rare cause of uterine bleeding. Diagnosis and treatment. AB - A case of uterine arteriovenous malformation following a dilatation and curettage is presented. Initial diagnosis with colour and pulsed Doppler ultrasound and treatment with transcatheter arterial embolization are described. PMID- 12890255 TI - Pelvic chondrosarcoma in an 18-year-old man masquerading as tuberculous ilipsoas abscess. AB - Chondrosarcoma is distinctly uncommon in the younger age group. We report an unusual presentation of pelvic bone chondrosarcoma in an 18-year-old man clinically creating an impression of tuberculous abscess, and we discuss the CT and MRI appearances. PMID- 12890256 TI - 'Benign' hepatic portal venous gas. AB - The presence of portal venous gas within the hepatic parenchyma is usually associated with a guarded prognosis and a mortality rate approaching 75%. However, there are infrequent causes of portal venous gas not associated with dire clinical outcomes. We describe three patients who made uneventful clinical recoveries after presenting with clinical and imaging manifestations of ischaemic bowel and hepatic portal venous gas, two of which had distended but non-necrotic bowel at laparotomy. PMID- 12890257 TI - Imaging features of primary pulmonary liposarcoma. AB - Primary liposarcoma of the lung is extremely rare. We report a 28-year-old pregnant woman who complained of dyspnoea during the third trimester. Chest radiography, thoracic ultrasound, CT and MRI showed a huge heterogeneous tumour involving all the left lung and the mediastinum. The tumour was composed of soft tissue, and fatty and cystic components with calcifications. Diagnosis was made on core biopsy under CT guidance. Surgical excision was performed but unfortunately the patient died during the operation. PMID- 12890258 TI - Gorlin's syndrome: diffuse appendicular skeletal involvement with scintigraphic correlation. AB - Gorlin's syndrome (also known as basal cell nevus syndrome, Gorlin-Goltz syndrome, and nevoid basal cell carcinoma syndrome) is a rare, inherited disorder characterized by multiple basal-cell epitheliomas, intracranial calcification, keratocysts of the mandible, and unusual and striking skeletal abnormalities. We present the interesting case of a 45-year-old woman who was informed that she had fibrous dysplasia of the extremity at another institution before extensive radiological work-up showed a diffuse skeletal process. The skeletal abnormalities, in conjunction with the patient's history of multiple basal cell carcinomas, is consistent with the diagnosis of Gorlin's syndrome. We describe this unusual case of striking radiological and scintigraphic findings in a patient with Gorlin's syndrome. PMID- 12890259 TI - Chondrosarcoma of bone complicating Ollier's disease: report of a favourable response to radiotherapy. AB - Because chondrosarcoma of bone is traditionally thought to be a radioresistant malignancy, it is usually managed surgically. We report a case of multifocal chondrosarcoma arising in Ollier's disease for which the patient declined surgery. He was given a course of radical radiotherapy that resulted in symptom palliation and a radiologically confirmed response before he died of disseminated disease. In patients with inoperable chondrosarcoma, radiotherapy can provide palliative benefit. PMID- 12890260 TI - Leptomeningeal carcinomatosis from squamous cell carcinoma of the supraglottic larynx. AB - Leptomeningeal carcinomatosis is an uncommon but devastating form of metastatic spread. To our knowledge, only 16 cases originating from a head and neck cancer have been reported. We describe the first case of a patient with leptomeningeal carcinomatosis arising from a laryngeal squamous cell carcinoma. Shortly after completing treatment for an advanced supraglottic laryngeal cancer, this 63-year old man presented with lower limb neurological symptoms and signs. Radiological and cytological evidence of leptomeningeal carcinomatosis of the distal spinal canal was identified. He was treated with intrathecal methotrexate and palliative radiotherapy. Although his pain improved, his lower limb weakness worsened. He died 3 weeks after completing radiotherapy. Presumed mode of spread was via the haematogenous route. The natural history and management of leptomeningeal carcinomatosis are discussed. Clinicians should be aware of the uncommon possibility of leptomeningeal carcinomatosis in a patient presenting with an appropriate constellation of symptoms and signs, and a past history of cancer. PMID- 12890263 TI - Case quiz. A pressing problem: posterior reversible leucenencephalopathy syndrome. PMID- 12890264 TI - Case quiz. Thoracic CT illustrating hyperdense bronchial mucous plugging: allergic bronchopulmonary aspergillosis. PMID- 12890265 TI - Radiology by non-radiologists. PMID- 12890266 TI - RE: Guide to the radiology report. PMID- 12890269 TI - Diagnosing eating disorders -- AN, BN and the others. PMID- 12890270 TI - Gender differences in unipolar depression: an update of epidemiological findings and possible explanations. AB - OBJECTIVE: To give an update on epidemiological findings on sex differences in the prevalence of unipolar depression and putative risk factors. MATERIAL AND METHODS: Systematic review of the literature. RESULTS: Recent epidemiological research yields additional evidence for a female preponderance in unipolar depression, holding true across different cultural settings. Current explanations include artefacts, genetic, hormonal, psychological and psychosocial risk factors. Rather consistently, intrapsychic and psychosocial gender role related risk factors have been identified which may contribute to the higher depression risk in women. Gender role aspects are also reflected in endocrine stress reactions and possibly influence associated neuropsychological processes. CONCLUSION: There is a need for more integrative models taking into account psychological, psychosocial, and macrosocial risk factors as well as their interactions, which also connect these factors with physiological and endocrine responses. Furthermore, it is conceivable that across the life span, as well as across cultural settings, individual risk factors will add with varying emphasis to the higher prevalence of depression in women. PMID- 12890271 TI - A critical examination of the amenorrhea and weight criteria for diagnosing anorexia nervosa. AB - BACKGROUND: Amenorrhea and weight loss to below 15% of 'healthy' weight are current diagnostic criteria for anorexia nervosa (AN). This study compares females who meet current International Statistical Classification of Diseases and Related Health Problems, tenth revision (ICD-10) and Diagnostic and Statistical Manual of Mental Disorders, fourth edition (DSM-IV) criteria for AN with females who meet the essential psychopathology and significant self-starvation, but lack either amenorrhea or weight loss below 85%. METHOD: This study retrospectively examined the medical charts of 588 consecutive admissions to an in-patient treatment program. All diagnoses conformed to ICD-10 and DSM-IV. RESULTS: Of 588 admissions, 297 females had some form of AN with 77.4% (230 of 297) meeting current criteria, while 22.6% (67 of 297) with core psychopathology and self starvation were classified as eating disorders not otherwise specified or 'atypical' because of some menstrual function or final weight above 85%. The groups showed few statistically significant differences on demographics, illness history, and treatment response, psychopathology, or bone density. CONCLUSION: Amenorrhea may not be a useful diagnostic criterion. Also, requiring below 85% of healthy weight may need generalization. PMID- 12890272 TI - Eating disorders and aggressiveness among adolescents. AB - OBJECTIVE: To explore the links between propensity to aggression and eating disorders in a sample of school-aged adolescents in a northeastern area of Italy. METHOD: In a mixed male-female sample of 1000 adolescents (10% of the district's population aged 15-19 years) we administered the Eating Attitudes Test, the Bulimic Investigatory Test of Edinburgh and the Body Attitudes Test, as measures of abnormal eating attitudes and behaviours, and the Aggression Questionnaire, as a measure of the propensity to aggression. RESULTS: Females scored significantly higher than males at all eating disorders inventories (P < 0.0001). Males scored higher than females at the Aggression Questionnaire. In both genders, there was a positive correlation between scores at any of the eating disorders inventories and those at the Aggression Questionnaire (P < 0.0001). CONCLUSION: Overtly expressed aggression might have a negative impact on the course of eating disorders and on the compliance with treatment, also enhancing the risk of suicide. PMID- 12890273 TI - Thyroid indices and treatment outcome in bulimia nervosa. AB - OBJECTIVE: This study examined the thyroxine (T4) and free T4 (FT4) status of women with bulimia nervosa and its value as a predictor of outcome. METHOD: A total of 135 women with bulimia nervosa underwent 12-weeks cognitive behavioral therapy treatment. Prior to and at 3-year follow-up patients completed psychiatric assessments and serumT4 and FT4 were measured. RESULTS: At 3-year follow-up, 71% had no eating disorder and 29% met criteria for any eating disorder diagnosis. Mean T4 and FT4 concentrations were within normal ranges. Pre treatment T4 and FT4 concentrations were inversely associated with food restriction and purging frequency, respectively. Compared with women with no eating disorder, those with any eating disorder at follow-up had lower pretreatment T4 concentrations. When pre-treatment food restriction, oral contraceptive use and binge frequency where controlled for, low T4 concentration was the only predictor of eating disorder diagnosis at follow-up. CONCLUSION: Low T4 concentrations at pretreatment may be a predictor of poor outcome in bulimia nervosa. PMID- 12890274 TI - Gender differences in binge-eating: a population-based twin study. AB - OBJECTIVE: To assess whether genetic and environmental effects on liability to binge-eating (BE) are of equal importance for males and females and whether the same genetic risk factors predispose to BE in the two sexes. METHOD: Questionnaire data on 8045 same sex and opposite sex twins, aged 19-31 years, from a population-based Norwegian registry, was used to estimate the relative contribution of genetic and environmental factors to liability for BE utilizing structural equation modeling. RESULTS: In the best-fitting model, the magnitude of genetic and environmental effects on BE was the same for males and females. Heritability was 51%. The correlation between genetic risk factors in men and women was estimated to be +0.57. CONCLUSION: Binge-eating appears to be equally heritable in males and females. Although the majority of the genetic risk factors are shared between the sexes, there may exist gender-specific genetic effects on liability. PMID- 12890275 TI - Comorbidity of impulse control disorders in pathological gamblers. AB - OBJECTIVE: To investigate the rate of impulse control disorders among pathological gamblers and examine the relationship of comorbidity to gambling severity. METHOD: Ninety-six adult pathological gamblers [mean age: 46.7 +/- 11.0 years; female: 44 (45.8%)] completed the following: Minnesota Impulsive Disorders Interview, Yale-Brown Obsessive-Compulsive Scale Modified for Pathological Gambling, and Gambling Symptom Assessment Scale. RESULTS: Twenty-two subjects (22.9%) reported a comorbid impulse control disorder, most commonly compulsive sexual behaviour and compulsive buying. Subjects with comorbidity reported significantly greater intensity of urges (t = -2.021; df = 94; P = 0.046) and thoughts (t = -2.147; df = 42.3; P = 0.038) related to gambling, and greater interference (t = -3.913; df = 48.1; P < 0.001) and distress (t = -2.504; df = 52.7; P = 0.015) secondary to gambling urges and thoughts. CONCLUSION: Impulse control disorders appear common among pathological gamblers and are associated with more severe gambling symptoms. PMID- 12890276 TI - Serum leptin and cholesterol levels in schizophrenic patients with and without suicide attempts. AB - OBJECTIVE: Previous studies demonstrate a relationship between lipid metabolism and suicide or impulsive-aggressive behaviours. Leptin seems to be related with lipid metabolism. Therefore, the aim was to measure total serum cholesterol and leptin levels in 16 medication-free schizophrenic patients with and without suicide attempts and in 16 healthy controls. METHOD: Subjects were assessed by using Impulsivity Rating (IRS) and Modified Overt Aggression Scale (MOAS). RESULTS: The patients had lower total cholesterol and leptin levels in serum compared with the controls. Significantly lower total cholesterol and leptin levels were observed in patients who had attempted suicide compared with those who had not. The levels were observed to be low in violent attempters when compared with non-violent attempters. MOAS and IRS scores were negatively correlated with both cholesterol or leptin levels in patients. CONCLUSION: The results indicated that medication-free schizophrenic patients have statistically significant lower serum cholesterol and leptin levels compared with controls and the difference is obvious in suicide attempters compared with non-suicide attempters and in violent attempters than non-violent attempters. PMID- 12890277 TI - Serum lipid levels and suicide attempts. AB - OBJECTIVE: To determine whether a correlation exists between lower serum lipid concentrations and increased suicide risk. METHOD: Serum lipid profiles were pair matched for 60 patients who had recently experienced failed attempts at suicide and equal numbers of non-suicidal psychiatric patients, and normal controls. Suicide attempt severity was scored using Weisman and Worden's risk-rescue rating scale. RESULTS: (a). Total serum cholesterol and low density lipoprotein levels were found to be lower in the parasuicidal population at statistically significant levels (P < 0.01 and <0.05, respectively); (b). triglyceride concentrations were lower in suicide attempters with major depression compared with non-suicidal depressed patients; and (c). risk-rescue rating scores were negatively correlated with total serum cholesterol levels (r = -0.347, P = 0.007). CONCLUSION: Low lipid metabolism may be a potential biological marker in the assessment of suicide risk. Further investigations are necessary to elucidate the biological mechanisms of these findings. PMID- 12890278 TI - Diagnostic classification of organic psychiatric disorders after aneurysmal subarachnoid hemorrhage: a comparison between ICD-10, DSM-IV and the Lindqvist & Malmgren classification system. AB - OBJECTIVE: There is no universally accepted consensus for organic psychiatric disorders (OPDs) between the two major classifications, ICD-10 and DSM-IV. The aim was to compare the coverage of these systems with the Lindqvist & Malmgren (LM) classification system for organic psychiatry. METHOD: Organic psychiatric disorders were diagnosed according to ICD-10, DSM-IV, and the LM system in 119 patients 12 months after surgery as a result of aneurysmal subarachnoid hemorrhage. RESULTS: Among 35 patients with astheno-emotional (AE) disorder (LM system), 83% (95% CI: 67-92%) had mild cognitive disorder (MCD) according to ICD 10 clinical descriptions and diagnostic guidelines (CDDG), 49% (95% CI: 33-64%) had MCD according to ICD-10 diagnostic criteria for research (DCR), and 34% (95% CI: 21-51%) had mild neurocognitive disorder according to DSM-IV. The coverage for other OPDs did not differ between the systems. CONCLUSION: The coverage for AE disorder (LM system) was significantly higher than the corresponding diagnoses of the ICD-10 and DSM-IV systems. Modifications of the latter systems are suggested. PMID- 12890279 TI - Psychological effects of the November 1999 earthquake in Turkey: an epidemiological study. AB - OBJECTIVE: This study assessed the traumatic stress symptoms and related factors in two towns affected by two earthquakes, which killed 20000 people in 1999 in Turkey. METHOD: A total of 430 people in selected households were seen 18 months after the earthquake. They were given a self-report questionnaire assessing post traumatic stress (PTSD) and depressive symptoms, demographics and trauma exposure. RESULTS: The rates of PTSD and depression were higher in the site closer to the epicenter. The traumatic stress symptom checklist scores were predicted by fear during earthquake, loss of friends and neighbours, female gender, lower education and living in rented accomodation. Depression was predicted by study site, death of relatives and past psychiatric illness. CONCLUSION: These results show that severe earthquakes can cause long-lasting morbidity. Our previous findings that showed a differential prediction for depressive and traumatic stress symptoms after earthquakes are also supported. PMID- 12890280 TI - Examination stress in Singapore primary schoolchildren: how compliance by subjects can impact on study results. AB - OBJECTIVE: Examinations are anecdotally viewed as extremely stressful to Singapore schoolchildren. We test this postulate by obtaining parental ratings of children's emotional stress levels longitudinally in a large representative sample of sixth (P6) and fifth (P5) class primary schoolchildren, respectively, exposed and unexposed to a streaming examination. METHOD: Children's stress levels were rated monthly by a parent for 10 months. RESULTS: Analyses failed to find evidence of any differential stress impact across P6 and P5 comparison groups, apart from a subset of P6 children whose parents complied with every monthly survey. CONCLUSION: The streaming examination in the final year of primary school did not emerge as a general stressor to children, but achieved salience within a defined subset of children whose parents were highly study compliant. Study compliance may be a proxy variable of some import, and have wider relevance to other cohort studies and to intervention trials. PMID- 12890281 TI - Total plasma l-tryptophan, free l-tryptophan and competing amino acid levels in a homicidal male adolescent with conduct disorder. AB - OBJECTIVE: Several studies have shown abnormal findings in human serotonin metabolism, such as increased total plasma l-tryptophan and free l-tryptophan levels among habitually violent antisocial offenders. It is not clear if these increased l-tryptophan levels are associated with adult antisocial personality disorder (ASP) or history of substance abuse, or if these levels are already present in adolescent subjects with conduct disorder (CD). METHOD: Total plasma and free l-tryptophan and competing amino acids (CAAs) were measured in a 15-year old adolescent offender, who was convicted for two homicides, and in 10 healthy male controls of similar age and body mass index (BMI). RESULTS: In the juvenile offender, plasma total l-tryptophan/CAA was 84% and free l-tryptophan/CAA 143% higher than average mean among controls. CONCLUSION: From this very aggressive boy with CD, findings of free l- and total l-tryptophan/CAA values were similar to those of habitually violent adult ASP offenders. As severe CDs in adolescence tend to develop into adults with ASP, increased l-tryptophan/CAA and free l tryptophan/CAA values may serve as early indicators for the development of habitually violent adult offenders. PMID- 12890284 TI - Desmoid tumors of the abdominal wall: A case report. AB - BACKGROUND: Desmoid tumors are slow growing deep fibromatoses with aggressive infiltration of adjacent tissue but without any metastatic potential. CASE PRESENTATION: We report on two female patients with desmoid tumor of the abdominal wall who underwent primary resection. Both patients had a history of an earlier abdominal surgery. Preoperative evaluation included abdominal ultrasound, magnetic resonance imaging and computed tomography. The histology in both cases revealed a desmoid tumor. CONCLUSION: Complete surgical resection is the first line management of this tumor entity. PMID- 12890285 TI - Prevalence of pesticide exposure in young males (2400 hours were considered as 'exposed.' The 2400 hour cut-off value was chosen on the basis of previous reports indicating that this figure represents heavy exposure to genotoxic agents. Statistical analysis was obtained using SPSS-10ledR;. Between 1991 and 2001, 61 young males with adenocarcinoma of the prostate were identified, of whom 56 patients with a mean age of 47 years (range: 40-49) had complete records of treatment and could be contacted for completion of the questionnaire. The most common stage at presentation was Stage III and the mean Gleason's score was 7.5 (range 5-9). Interestingly, almost a third (16/56, 28.6%) of patients had stage IV disease at presentation. 37/56 (66.1%) patients had 'significant' exposure in our study. In addition, interestingly, the mean survival in the subgroup of patients with pesticide exposure was 11.3 months (SD: +/- 2.3 months), while the mean survival in the patients without pesticide exposure (n = 19) was 20.1 months (SD: +/- 3.1 months), with p-value <0.01. Although our study is relatively small, it does reveal preliminary evidence linking pesticide exposure to the early development of, possibly aggressive, prostate adenocarcinoma. Future, larger, epidemiological studies are needed to confirm the findings of our study. PMID- 12890286 TI - A Viral Platform for Chemical Modification and Multivalent Display. AB - The ability to chemically modify the surfaces of viruses and virus-like particles makes it possible to confer properties that make them potentially useful in biotechnology, nanotechnology and molecular electronics applications. RNA phages (e.g. MS2) have characteristics that make them suitable scaffolds to which a variety of substances could be chemically attached in definite geometric patterns. To provide for specific chemical modification of MS2's outer surface, cysteine residues were substituted for several amino acids present on the surface of the wild-type virus particle. Some substitutions resulted in coat protein folding or stability defects, but one allowed the production of an otherwise normal virus-like particle with an accessible sulfhydryl on its surface. PMID- 12890287 TI - Challenge of the world order and its implications for health personnel. PMID- 12890288 TI - PCR and Mosquito dissection as tools to monitor filarial infection levels following mass treatment. AB - BACKGROUND: Entomological methods may provide important tools for monitoring the progress of lymphatic filariasis elimination programs. In this study, we compared dissection of the vector, Culex quinquefasciatus, with the polymerase chain reaction (PCR) to assess filarial infection levels in mosquitoes in the context of a lymphatic filariasis elimination program in Leogane, Haiti. METHODS: Mosquitoes were collected using gravid traps located in 4 sentinel communities with Wuchereria bancrofti microfilaria prevalence that ranged from 0.8% to 15.9%. Captured mosquitoes were divided between dissection, to enumerate W. bancrofti larvae (L1, L2, L3) and desiccation for later analysis by PCR. PCR was conducted on DNA extracts from pooled mosquitoes (1-15 pooled females) utilizing a competitive PCR system with primers specific for the Ssp I repeat. PCR products were analyzed with a hybridization ELISA using probes specific for a control sequence and the Ssp I repeat. RESULTS: The prevalence of mosquito infection with W. bancrofti ranged from 0%-3.66% by dissection (L1-L3) and point estimates of infection prevalence, as assayed by PCR, ranged from 0.25% - 9.16%. Following mass treatment, W. bancrofti infection prevalence dropped significantly as determined by PCR and dissection in 2 of the 4 sentinel sites (Leogane and Barrier Jeudi, P = 0.04 and P = 0.005, respectively). Although transmission declined in the other two sites, larval recoveries were low and these changes were not statistically significant. DISCUSSION: Our results suggest that a single round of mass treatment can have an impact on transmission of lymphatic filariasis. The use of entomologic methods as a tool to monitor filariasis programs and the statistical limitations of mosquito trapping are discussed. PMID- 12890289 TI - 17beta-Estradiol responsiveness of MCF-7 laboratory strains is dependent on an autocrine signal activating the IGF type I receptor. AB - BACKGROUND: Human MCF-7 cells have been studied extensively as a model for breast cancer cell growth. Many reports have established that serum-starved MCF-7 cells can be induced to proliferate upon the sole addition of 17beta-estradiol (E2). However, the extent of the mitogenic response to E2 varies in different MCF-7 strains and may even be absent. In this study we compared the E2-sensitivity of three MCF-7 laboratory strains. RESULTS: The MCF-7S line is non-responsive to E2, the MCF-7 ATCC has an intermediate response to E2, while the MCF-7 NKI is highly E2-sensitive, although the levels and activities of the estrogen receptor (ER) are not significantly different. Both suramin and IGF type I receptor blocking antibodies are able to inhibit the mitogenic response to E2-treatment in MCF-7 ATCC and MCF-7 NKI cells. From this we conclude that E2-induced proliferation is dependent on IGF type I receptor activation in all three MCF-7 strains. CONCLUSIONS: The results presented in this article suggest that E2-responsiveness of MCF-7 cells is dependent on the secretion of an autocrine factor activating the IGF-IR. All three strains of MCF-7 breast cancer cells investigated do not respond to E2 if the IGF-RI-pathway is blocked. Generally, breast cancer therapy is targeted at inhibiting estrogen action. This study suggests that inhibition of IGF-action in combination with anti-estrogen-treatment may provide a more effective way in treatment or even prevention of breast cancer. PMID- 12890290 TI - Metabolic and haemodynamic effects of oral glucose loading in young healthy men carrying the 825T-allele of the G protein beta3 subunit. AB - BACKGROUND: A C825T polymorphism was recently identified in the gene encoding the beta3 subunit of heterotrimeric G-proteins (GNB3). The T-allele is significantly associated with essential hypertension and obesity. In order to further explore a possible pathogenetic link between the T-allele and impaired glucose tolerance we studied metabolic and haemodynamic responses to oral glucose loading in young, healthy subjects with and without the 825T-allele. METHODS: Twelve subjects with and 10 without the 825T-allele were investigated at rest and following glucose ingestion (75 g). Blood glucose, serum insulin and haemodynamics were determined prior to and over 2 hours following glucose ingestion. We non-invasively measured stroke volume (SV, by impedance-cardiography), blood pressure (BP), heart rate (HR), and systolic-time-intervals. Cardiac output (CO) was calculated from HR and SV. Total peripheral resistance was calculated from CO and BP. Metabolic and haemodynamic changes were quantified by maximal responses and by calculation of areas under the concentration time profile (AUC). Significances of differences between subjects with and without the T-allele were determined by unpaired two tailed t-tests. A p < 0.05 was considered statistically significant. RESULTS: Metabolic and haemodynamic parameters at baseline were very similar between both groups. The presence of the T-allele did not alter the response of any metabolic or haemodynamic parameter to glucose loading. CONCLUSIONS: In conclusion, this study does not support the hypothesis that the C825T polymorphism may serve as a genetic marker of early impaired glucose tolerance. PMID- 12890291 TI - Preventive medical care in remote Aboriginal communities in the Northern Territory: a follow-up study of the impact of clinical guidelines, computerised recall and reminder systems, and audit and feedback. AB - BACKGROUND: Interventions to improve delivery of preventive medical services have been shown to be effective in North America and the UK. However, there are few studies of the extent to which the impact of such interventions has been sustained, or of the impact of such interventions in disadvantaged populations or remote settings. This paper describes the trends in delivery of preventive medical services following a multifaceted intervention in remote community health centres in the Northern Territory of Australia. METHODS: The intervention comprised the development and dissemination of best practice guidelines supported by an electronic client register, recall and reminder systems and associated staff training, and audit and feedback. Clinical records in seven community health centres were audited at regular intervals against best practice guidelines over a period of three years, with feedback of audit findings to health centre staff and management. RESULTS: Levels of service delivery varied between services and between communities. There was an initial improvement in service levels for most services following the intervention, but improvements were in general not fully sustained over the three year period. CONCLUSIONS: Improvements in service delivery are consistent with the international experience, although baseline and follow-up levels are in many cases higher than reported for comparable studies in North America and the UK. Sustainability of improvements may be achieved by institutionalisation of relevant work practices and enhanced health centre capacity. PMID- 12890292 TI - The pharmacokinetics of the interstitial space in humans. AB - BACKGROUND: The pharmacokinetics of extracellular solutes is determined by the blood-tissue exchange kinetics and the volume of distribution in the interstitial space in the different organs. This information can be used to develop a general physiologically based pharmacokinetic (PBPK) model applicable to most extracellular solutes. METHODS: The human pharmacokinetic literature was surveyed to tabulate the steady state and equilibrium volume of distribution of the solutes mannitol, EDTA, morphine-6-glucuronide, morphine-3-glucuronide, inulin and beta-lactam antibiotics with a range of protein binding (amoxicillin, piperacillin, cefatrizine, ceforanide, flucloxacillin, dicloxacillin). A PBPK data set was developed for extracellular solutes based on the literature for interstitial organ volumes. The program PKQuest was used to generate the PBPK model predictions. The pharmacokinetics of the protein (albumin) bound beta lactam antibiotics were characterized by two parameters: 1) the free fraction of the solute in plasma; 2) the interstitial albumin concentration. A new approach to estimating the capillary permeability is described, based on the pharmacokinetics of the highly protein bound antibiotics. RESULTS: About 42% of the total body water is extracellular. There is a large variation in the organ distribution of this water - varying from about 13% of total tissue water for skeletal muscle, up to 70% for skin and connective tissue. The weakly bound antibiotics have flow limited capillary-tissue exchange kinetics. The highly protein bound antibiotics have a significant capillary permeability limitation. The experimental pharmacokinetics of the 11 solutes is well described using the new PBPK data set and PKQuest. CONCLUSIONS: Only one adjustable parameter (systemic clearance) is required to completely characterize the PBPK for these extracellular solutes. Knowledge of just this systemic clearance allows one to predict the complete time course of the absolute drug concentrations in the major organs. PKQuest is freely available http://www.pkquest.com. PMID- 12890293 TI - Habitual prospective memory in schizophrenia. AB - BACKGROUND: Prospective memory (PM), the act of remembering that something has to be done in the future without any explicit prompting to recall, provides a useful framework with which to examine problems in internal-source monitoring. This is because it requires distinguishing between two internally-generated processes, namely the intention to perform an action versus actual performance of the action. In habitual tasks, such as taking medicine every few hours, the same PM task is performed regularly and thus it is essential that the individual is able to distinguish thoughts (i.e., thinking about taking the medicine) from actions (i.e., actually taking the medicine). METHODS: We assessed habitual PM in patients with schizophrenia by employing a laboratory analogue of a habitual PM task in which, concurrently with maneuvering a ball around an obstacle course (ongoing activity), participants were to turn over a counter once during each trial (PM task). After each trial, participants were asked whether they had remembered to turn the counter over. RESULTS: Patients with schizophrenia made a disproportionate number of errors compared to controls of reporting that a PM response had been made (i.e., the counter turned over) after an omission error (i.e., the counter was not turned over). There was no group difference in terms of reporting that an omission error occurred (i.e., forgetting to turn over the counter) when in fact a PM response had been made. CONCLUSION: Patients with schizophrenia displayed a specific deficit distinguishing between two internally generated sources, attributable to either poor source monitoring or temporal discrimination. PMID- 12890294 TI - Description and evaluation of a delivery system for aerosolized prostacyclin. AB - INTRODUCTION: Inhaled vasodilators such as nitric oxide and aerosolized prostacyclin (PGI(2)) are used to treat severe hypoxemia in acute respiratory distress syndrome. Preferential distribution of nitric oxide and PGI(2) to ventilated areas of the lung causes selective pulmonary vasodilation, improved ventilation/perfusion matching, and decreased hypoxemia. Because of the technical limitations of previously described methods, we developed a PGI(2) delivery technique that allows the aerosolized drug dose to be easily calculated, set, and adjusted. METHODS: A 50 mL solution of PGI(2) (3.0x10(4) ng/mL) and a 500 mL normal saline solution were infused by a dual-channel volumetric infusion pump into a MiniHEART jet nebulizer that has a manufacturer-specified output of 8 mL/h at a set flow of 2 L/min. By adjusting the pump infusion rate to achieve a total output of 8 mL/h, the PGI(2) concentration was altered to deliver a calculated aerosolized dose of 10-50 ng/kg/min. The effectiveness of the delivery system was retrospectively evaluated by way of the responses of 11 severely hypoxemic acute respiratory distress syndrome patients who received PGI(2) via the system we describe. The MiniHEART nebulizer output, particle size, and dose delivery were evaluated in a laboratory bench study, using a set flow of 2 L/min. RESULTS: Aerosolized PGI(2) therapy (mean dose 28 +/- 17 ng/kg/min, range 10-50 ng/kg/min) significantly increased the ratio of P(aO)(2) to fraction of inspired oxygen (P(aO)(2)/F(IO)(2)) (60 +/- 11 mm Hg vs 80 +/- 17 mm Hg, p = 0.003) and arterial oxygen saturation measured via pulse oximetry (86 +/- 8% vs 94 +/- 3%, p = 0.005) (differences evaluated with the Wilcoxon signed rank test). There was no difference in positive end-expiratory pressure, mean airway pressure, or F(IO)(2), before and after aerosolized PGI(2) (p > 0.05). Nebulizer output was 6.8 +/- 0.9 mL/h, range 6.0-7.8 mL/h. The inhaled aerosol particles had a mass median diameter of 3.1 micro m. Emitted dose was 67 +/- 13% (range 57-81%) of the calculated dose. CONCLUSION: Our system is effective in delivering aerosolized PGI(2) to the alveolar-capillary interface, as indicated by significant oxygenation improvements soon after therapy commenced. The performance of the MiniHEART nebulizer varies from the manufacturer's specifications, which may alter the delivered dose. PMID- 12890295 TI - Effects of expiratory rib cage compression and/or prone position on oxygenation and ventilation in mechanically ventilated rabbits with induced atelectasis. AB - INTRODUCTION: Expiratory rib cage compression is a chest physiotherapy technique known as "squeezing" technique in Japan. It has been claimed that rib cage compression effectively treats and/or prevents lung collapse, but no studies have been reported on rib cage compression focused on improving ventilation and/or oxygenation in subjects with collapsed lung. Therefore, we studied whether rib cage compression, with and without prone positioning, improves the ratio of P(aO)(2) to fraction of inspired oxygen (P(aO)(2)/F(IO)(2)), P(aCO)(2), or dynamic compliance of the respiratory system. METHODS: We used anesthetized adult rabbits with induced atelectasis. An endotracheal tube and an 18-gauge catheter were placed into the airway via a tracheostoma, and pressure-controlled mechanical ventilation was used. To create atelectasis, artificial mucus was infused into the airway via the catheter. The rabbits were randomly assigned to one of 4 groups ( = 10 in each group): (1) supine without rib cage compression, (2) supine with rib cage compression, (3) prone without rib cage compression, and (4) prone with rib cage compression. Each rib cage compression session lasted for 5 min and was repeated 5 times every 30 min. After these interventions for 180 min all animals were placed in the supine position for 120 min. RESULTS: The prone-position groups had significantly higher P(aO)(2)/F(IO)(2) than the supine position groups at 60 min after the beginning of the intervention, and at 60, 90, and 120 min after the end of the intervention (p < 0.05). Rib cage compression did not significantly affect P(aO)(2)/F(IO)(2), P(aO)(2), or dynamic compliance. CONCLUSIONS: It is unlikely that rib cage compression re-expands collapsed lung. Prone positioning improved oxygenation in rabbits with induced atelectasis. PMID- 12890296 TI - Standards and interpretive issues in lung function testing. AB - Pulmonary function tests are most useful when performed with good technique and with an accurate system. Using standard techniques in performing the tests minimizes diagnostic and therapeutic errors. This report discusses the rationale and limits of standardization and offers practical suggestions for using available standards to increase confidence in test results. PMID- 12890297 TI - How to make sure your spirometry tests are of good quality. AB - Poorly performed spirometry greatly increases the risk of misinterpreting spirometry results. The most common cause of erroneous results is suboptimal patient coaching. Use exaggerated body language to demonstrate each of the 3 phases of the forced vital capacity (FVC) maneuver. The first phase of the maneuver (the maximally deep breath) is the most important and should receive the most emphasis. In the second phase (the blast) startle the patient to prompt maximum flow during the first second. In the third phase do not yell at the patient to keep blowing; instead, draw the patient's attention to the movement of the bell of the volume spirometer, the computer incentive display, or the audio tone of the flow-sensing spirometer, which shows that he or she is continuing to get some air out. Pay attention to the patient's body language as you coach him or her through the 3 phases. Facial expressions and body language are much more important than telling the patient what to do. Use the latest National Health and Nutrition Examination Survey (NHANES III) reference equations and the ratio of forced expiratory volume in the first second to forced expiratory volume in the first 6 seconds (FEV(1)/FEV(6)). Young, old, and sick patients can produce high quality, reproducible pulmonary function test results. Grade pulmonary function test efforts with the scholastic grading system (A, B, C, D, and F). Implement a centralized spirometry quality assurance program. Test your spirometers daily. Be cautious in making corrections for body-temperature-and-pressure-saturated. PMID- 12890298 TI - Diffusing capacity: how to get it right. AB - The carbon monoxide diffusing capacity test (D(LCO)) is a commonly performed pulmonary function test that requires technical expertise and attention to detail to get acceptable results. With the advent of automated devices and powerful computer programs, D(LCO) measurement has rapidly gained wide clinical acceptance. But there are many subtle aspects to performing the test that can diminish its accuracy and repeatability. The clinician must ensure: that the D(LCO) instrument is correctly calibrated; that inhalation is least 90% of the largest previously measured vital capacity; that the patient executes a quick, smooth inhalation within 2 seconds; that the breath-hold is 9-11 seconds; that the breath-hold is without straining (no Valsalva or Muller maneuvers); that exhalation is quick and smooth; that a representative gas sample is obtained from the correct portion of the exhalation; and that at least 5 minutes elapse between D(LCO) tests. At least 2 but no more than 5 D(LCO) tests should be conducted, and testing is complete when 2 tests are within 10% or 3 D(LCO) units (mL CO/min/mm Hg) of each other. The reported D(LCO) value is the average of the first 2 tests that meet the reproducibility criteria, but if 5 tests are performed and no 2 meet the reproducibility criteria, the reported value is the average of the 2 tests with the highest inspiratory volumes. These quality controls will help laboratories achieve consistent high D(LCO) accuracy. PMID- 12890299 TI - The six-minute walk test. AB - The American Thoracic Society has issued guidelines for the 6-minute walk test (6MWT). The 6MWT is safer, easier to administer, better tolerated, and better reflects activities of daily living than other walk tests (such as the shuttle walk test). The primary measurement is 6-min walk distance (6MWD), but during the 6MWT data can also be collected about the patient's blood oxygen saturation and perception of dyspnea during exertion. When conducting the 6MWT do not walk with the patient and do not assist the patient in carrying or pulling his or her supplemental oxygen. The patient should walk alone, not with other patients. Do not use a treadmill on which the patient adjusts the speed and/or the slope. Do not use an oval or circular track. Use standardized phrases while speaking to the patient, because your encouragement and enthusiasm can make a difference of up to 30% in the 6MWD. Count the laps with a lap counter. If the 6MWD is low, thoroughly search for the cause(s) of the impairment. Better 6MWD reference equations will be published in the future, so be sure you are using the best available reference equations. PMID- 12890300 TI - Pulmonary function testing: coding and billing issues. AB - Clinicians who conduct pulmonary function tests should understand the principles and rules of the coding and billing system for pulmonary function testing. Certain billing codes will not be paid by most insurance payers. To ensure that your pulmonary function tests are appropriately coded, billed, and paid: (1) obtain a Current Procedural Terminology (CPT) coding book and an International Classification of Diseases 9th Revision (ICD-9) diagnosis book, and understand how they are used in setting coding and billing strategies, (2) know the people in your facility who do the billing and work with them to produce an appropriate coding and billing strategy, (3) make sure the physicians are involved in developing and implementing your coding and billing strategy, and (4) assure that your laboratory is set up properly to follows the Medicare rules for participation, that you have the appropriate testing supervision, that the appropriate administrative structure is in place to assure compliance with all regulations, and that you meet American Thoracic Society testing standards. PMID- 12890301 TI - Olanzapine orally disintegrating tablets in the treatment of acutely ill non compliant patients with schizophrenia. AB - The objective of this study was to determine if the orally disintegrating tablet formulation of olanzapine, Zyprexa Zydis, would facilitate antipsychotic medication compliance in acutely ill, non-compliant patients. Eighty-five acutely ill patients with schizophrenia or schizoaffective disorder who met medication non-compliance criteria received open-label olanzapine orally disintegrating tablets (1020 mgd) for up to 6 wk. Improvement in medication compliance was assessed using various rating scales to measure changes in psychopathology, medication-taking and compliance attitudes, and nursing care burden. Safety variables were also measured. Significant improvement from baseline was demonstrated in the Positive and Negative Syndrome Scale total score at Week 1 and subsequently (p0.001). Significant improvement from baseline was also seen in various scales measuring medication compliance, attitude, and nursing care burden (p0.05). Olanzapine orally disintegrating tablets were well-tolerated. Olanzapine orally disintegrating tablets may benefit acutely ill, non-compliant schizophrenic patients by facilitating acceptance of active antipsychotic drug therapy. PMID- 12890302 TI - Lack of effect of amisulpride on the pharmacokinetics and safety of lithium. AB - Lithium may be used as adjuvant therapy in schizophrenic patients and antipsychotics can be employed during the early phases of lithium therapy in patients with bipolar disorder. The issue of interactions between lithium and antipsychotics is therefore important. This study investigates the potential influence of repeated administration of amisulpride, an atypical antipsychotic, on the pharmacokinetics of lithium at steady state. Twenty-four healthy male volunteers (aged 1833 yr) received lithium carbonate (500 mg b.i.d.) for 14 d. All subjects were shown to have stable lithium serum concentrations after 57 d and were then randomized to receive double-blind administration of amisulpride (100 mg b.i.d.) or placebo bid from day 8 of lithium administration. Complete pharmacokinetic profiles were obtained on days 7 and 14 for lithium and trough plasma concentrations on days 10, 12 and 14 for amisulpride. Co-administration of amisulpride appeared to exert no effect on the pharmacokinetics of lithium. All treatments were well tolerated and safety assessment revealed no differences between the lithium+placebo and lithium+amisulpride groups. This finding permits the flexible use of amisulpride in patients already receiving lithium therapy. PMID- 12890303 TI - Delusions of pregnancy associated with increased prolactin concentrations produced by antipsychotic treatment. AB - Treatment of psychotic symptoms has traditionally involved conventional antipsychotics. While efficacious, their side-effects have been problematic and the approval by the Food and Drug Administration of the newer antipsychotics with improved side-effects profiles heralded important advances in treating psychoses. Prolactin elevation has been associated with all classical and some atypical antipsychotics. We present cases where elevation of prolactin concentrations secondary to antipsychotic treatment was associated with delusions of pregnancy. Risperidone was the antipsychotic employed and elevation of prolactin concentrations were noted each time. The delusions abated and prolactin concentrations decreased when the drug was discontinued. Rechallenge with risperidone resulted in re-elevation of prolactin levels along with recurrent delusions. Substituting risperidone with another antipsychotic (either olanzapine or quetiapine) also led to abatement of the delusions and lowering of prolactin. Although no direct psychotogenic effects of prolactin are known, it is contended that delusions of pregnancy reported during antipsychotic treatment might be associated with rising prolactin concentrations. PMID- 12890304 TI - Lack of effect of mood stabilizers or neuroleptics on GSK-3 protein levels and GSK-3 activity. AB - Glycogen synthase kinase (GSK)-3 protein levels and GSK-3 activity were previously found to be over 40% reduced in the post-mortem prefrontal cortex of schizophrenic patients. Lithium and valproate have been reported to selectively inhibit GSK-3. We hypothesized that in-vivo administration of lithium and valproate would result in up-regulation of GSK-3 protein levels and GSK-3 activity. The present study aimed to evaluate the possible involvement of neuroleptic treatment in the decrease of GSK-3 in schizophrenia. Rat frontal cortex GSK-3 protein levels and GSK-3 activity were measured following administration of therapeutic doses of lithium or valproate for 11 d, or of haloperidol, chlorpromazine or clozapine for 21 d. None of the drugs induced a change in GSK-3 protein levels. All the drugs except chlorpromazine (which was not tested) did not affect GSK-3 activity. This suggests that GSK-3 inhibition by lithium or valproate does not induce regulation of protein levels or activity and that the reduction in GSK-3 protein levels and GSK-3 activity in the post-mortem prefrontal cortex of schizophrenic patients is not neuroleptic-treatment related. PMID- 12890305 TI - Introduction to the Special Section. New advances in the understanding and treatment of bipolar disorder. AB - The current renewed interest in the field of bipolar disorders can be attributed to various factors which are reviewed in this Special Section of the International Journal of Neuropsychopharmacology. Bipolar disorders are one of the most frequent disorders in psychiatry and this Special Section presents papers on recent advances in the clinical course, epidemiology, biological models and neurobiology of bipolar disorders. Novel therapeutic approaches will be covered in a second part in Volume 6, Number 3 (September 2003). PMID- 12890306 TI - Long-term symptomatic status of bipolar I vs. bipolar II disorders. AB - Weekly affective symptom severity and polarity were compared in 135 bipolar I (BP I) and 71 bipolar II (BP II) patients during up to 20 yr of prospective symptomatic follow-up. The course of BP I and BP II was chronic; patients were symptomatic approximately half of all follow-up weeks (BP I 46.6% and BP II 55.8% of weeks). Most bipolar disorder research has concentrated on episodes of MDD and mania and yet minor and subsyndromal symptoms are three times more common during the long-term course. Weeks with depressive symptoms predominated over manichypomanic symptoms in both disorders (31) in BP I and BP II at 371 in a largely depressive course (depressive symptoms=59.1% of weeks vs. hypomanic=1.9% of weeks). BP I patients had more weeks of cyclingmixed polarity, hypomanic and subsyndromal hypomanic symptoms. Weekly symptom severity and polarity fluctuated frequently within the same bipolar patient, in which the longitudinal symptomatic expression of BP I and BP II is dimensional in nature involving all levels of affective symptom severity of mania and depression. Although BP I is more severe, BP II with its intensely chronic depressive features is not simply the lesser of the bipolar disorders; it is also a serious illness, more so than previously thought (for instance, as described in DSM-IV and ICP-10). It is likely that this conventional view is the reason why BP II patients were prescribed pharmacological treatments significantly less often when acutely symptomatic and during intervals between episodes. Taken together with previous research by us on the long-term structure of unipolar depression, we submit that the thrust of our work during the past decade supports classic notions of a broader affective disorder spectrum, bringing bipolarity and recurrent unipolarity closer together. However the genetic variation underlying such a putative spectrum remains to be clarified. PMID- 12890307 TI - Bipolar comorbidity: from diagnostic dilemmas to therapeutic challenge. AB - Comorbidity in bipolar disorder is the rule rather than the exception more than 60% of bipolar patients have a comorbid diagnosis and is associated with a mixed affective or dysphoric state; high rates of suicidality; less favourable response to lithium and poorer overall outcome. There is convincing evidence that rates of substance use and anxiety disorders are higher among patients with bipolar disorder compared to their rates in the general population. The interaction between anxiety disorders and substance use goes both ways: patients with bipolar disorder have a higher rate of substance use and anxiety disorder, and vice versa. Bipolar disorder is also associated with borderline personality disorder and ADHD, and to a lesser extent with weight gain. As more than 40% of bipolar patients have anxiety disorder, it is indicated that while diagnosing bipolar patients, systematic enquiry about different anxiety disorders is called for. This also presents a therapeutic challenge, since agents that effectively treat anxiety disorders are associated with the risk of induced mania. Therefore, the treating psychiatrist needs to carefully evaluate the potential benefit of treating the anxiety against the potential cost of inducing a manic episode. A possible solution would be to use, when possible, a non-pharmacological intervention, such as a cognitivebehavioural approach. Alternately, it is suggested that the clinician attempts to ensure that the patient receives adequate treatment with mood stabilizers before slowly and carefully attempting the addition of anti-anxiety compounds with a relatively lower risk of mania induction (e.g. SSRIs compared to TCAs). PMID- 12890308 TI - Natural course and burden of bipolar disorders. AB - Despite an abundance of older and more recent retrospective and considerably fewer prospective-longitudinal studies in bipolar disorders I and II, there are still remarkable deficits with regard to our knowledge about the natural course and burden. The considerable general and diagnosis-specific challenges posed by the nature of bipolar disorders are specified, highlighting in particular problems in diagnostic and symptom assessment, shifts in diagnostic conventions and the broadening of the diagnostic concept by including bipolar spectrum disorders. As a consequence it still remains difficult to agree on several core features of bipolar disorders, such as when they begin, how many remit spontaneously and how many take a chronic course. On the basis of clinical and epidemiological findings this paper summarizes (i) a significant need to extend the study of the natural course of bipolar disorder in clinical samples beyond the snapshot of acute episodes to the study of the mid-term and long-term symptom course, associated comorbidities and the associated burden of the disease. (ii) In terms of epidemiological studies, that are also of key importance for resolving the critical issues of threshold definitions in the context of the bipolar spectrum concept, there is a clear need for identifying the most relevant risk factors for the first onset and those for the further illness progression in early stages. Since there are some indications that these critical processes might start as early as adolescence, such studies might concentrate on young cohorts and clearly before these prospective patients come to clinical attention. (iii) The value of both types of studies might be enhanced, if beyond the use of standardized diagnostic interview, special attempts are made to use prospective life- and episode-charting methods for bipolar illnesses. PMID- 12890309 TI - Molecular genetics of affective disorders. AB - Family studies have provided evidence for familial transmission in suicide in major psychiatric disorders, and in particular affective disorders. Even though they may seem contradictory, linkage studies have suggested several genetic regions implicated in affective disorders. Association studies have mainly focused on genes related to serotonergic and monoaminergic pathways. Other genes involved in GABAergic and substance P pathways have also been studied in association studies. Another way to approach the genetics of affective disorders is the definition of more detailed phenotypes. Suicidal behaviour is one of the more largely studied subphenotypes within affective disorders. Tryptophan hydroxylase and serotonin transporter genes, related to the serotonergic pathway, have been found to be associated to suicidal behaviour, in particular violent suicidal behaviour but need to be replicated before definitive conclusion. Improved methodologies and updated tools in genetic studies will improve in the future our knowledge of affective disorders. PMID- 12890310 TI - Contributions from brain imaging to the elucidation of pathophysiology of bipolar disorder. AB - Over the past two decades, brain-imaging studies have examined the mechanisms possibly involved in the pathophysiology of bipolar and unipolar mood disorders. The available findings suggest subtle anatomical changes in sub-regions of the prefrontal cortex, medial temporal lobe and cerebellum, and functional abnormalities in brain circuits inter-connecting these same brain regions and the striatum in patients suffering from bipolar disorder. 1H magnetic resonance spectroscopy (MRS) studies reported decreased N-acetyl aspartate (NAA) levels in the dorsolateral prefrontal cortex, and 31P-MRS studies found abnormalities in membrane phospholipids in frontal and temporal regions in bipolar individuals. Few studies have utilized in-vivo receptor imaging to study bipolar patients. Even though preliminary findings from cross-sectional studies indicate anatomical, neurochemical, and functional brain abnormalities in bipolar patients in key regions involved in mood regulation, the relationship of such abnormalities with illness phase and their clinical relevance needs further investigation. The potential for utilization of brain-imaging tools to elucidate the pathophysiology of bipolar disorder is still largely unrealized, and it is anticipated that important new developments in this area will come about over the next years and beyond. PMID- 12890311 TI - Cellular mechanisms and second messengers: relevance to the psychopharmacology of bipolar disorders. AB - The discovery of lithiums efficacy as a mood-stabilizing agent revolutionized the treatment of patients with bipolar disorder and after five decades, lithium continues to be the mainstay of treatment for bipolar disorder. Recent research on the molecular mechanism underlying the therapeutic effect of lithium has focused on how it changes the activities of cellular signal transduction systems, especially the cyclic AMP and phosphoinositide second-messenger systems. Considerable data suggest that carbamazepine and valproate (VPA) are an alternative or adjunctive treatment to lithium. VPA, despite being dissimilar structurally to lithium, shares most of the effects of lithium at the level of protein kinase C (PKC). Like lithium, VPA reduces the activity of PKC and reduces the protein levels of different PKC isoforms, however the effects of VPA appear to be largely independent of myoinositol. The long-term efficacy of VPA and lithium in bipolar disorder suggested that modulation of gene expression might be an important target for these drugs. Both VPA and lithium altered the expression of the early inducible genes for c-fos and c-jun thus promoting the expression of specific proteins. The genes known to be regulated by the AP-1 family of transcription factors include genes for various neuropeptides, neurotrophins, receptors, transcription factors, enzymes, proteins that bind to cytoskeletal elements, and cytoprotective proteins such as bcl-2. In conclusion chronic treatment with lithium and other mood stabilizers, by regulating transcriptional factors, may modulate the expression of a variety of genes that compensate for aberrant signalling associated with the pathophysiology of bipolar disorder. PMID- 12890312 TI - Neuroendocrine profiles in mood disorders. AB - The study of neuroendocrine abnormalities in major mental illness, such as the unipolar and bipolar affective syndromes, has been the focus of interest in the past few years. The neuroendocrine window into the brain has been considered as a fruitful and promising approach to the study of mental disorders, as suggested by studies of some neuroendocrine challenge tests in depression that demonstrated their potential use as biological markers. The modern approach to hormonal dynamics focuses on the circadian and pulsatile profiles that truly represent physiological modulation and tests the hypothesis that an abnormality in circadian rhythms is present in affective illness. From the fundamental point of view, such studies performed using a frequent sampling interval over the 24-h cycle aim to clarify the control and significance of the temporal sleep and wake fluctuations of neuroendocrine system activities. Twenty-four-hour hypersecretion of cortisol, diurnal hypersecretion of growth hormone, and normal 24-h levels of prolactin have been reported in careful chronobiological studies of depressed patients, along with sleep recordings. In addition, a nocturnal quiescent period, and a subsequent increase towards the morning maximum, have been consistently found in a subset of depressed patients suffering from endogenous depression. After successful treatment with antidepressants, most of these abnormalities (with the exception of those found in the prolactin study) tend to correct. The normalization of the timing of hormonal secretion was accompanied by a correction of sleep abnormalities and in particular, a lengthening of the REM latencies. Normalization of cortisol secretion was associated with a decrease in the magnitude of episodic cortisol pulses whereas normalization of growth hormone secretion was due to a diminished number of secretory pulses. In conclusion, a disorder of circadian time-keeping seems to characterize acute episodes of major endogenous depression in some patients. This abnormality as well as the associated increases in adrenocorticotropic and somatotropic activities seem to be a state-, rather than trait-dependent phenomenon. PMID- 12890313 TI - Quetiapine in a delusional depressed elderly patient: no EPS and a favourable outcome. PMID- 12890314 TI - Mycobacterial aerosols and respiratory disease. AB - Environmental opportunistic mycobacteria, including Mycobacterium avium, M. terrae, and the new species M. immunogenum, have been implicated in outbreaks of hypersensitivity pneumonitis or respiratory problems in a wide variety of settings. One common feature of the outbreaks has been exposure to aerosols. Aerosols have been generated from metalworking fluid during machining and grinding operations as well as from indoor swimming pools, hot tubs, and water damaged buildings. Environmental opportunistic mycobacteria are present in drinking water, resistant to disinfection, able to provoke inflammatory reactions, and readily aerosolized. In all outbreaks, the water sources of the aerosols were disinfected. Disinfection may select for the predominance and growth of mycobacteria. Therefore, mycobacteria may be responsible, in part, for many outbreaks of hypersensitivity pneumonitis and other respiratory problems in the workplace and home. PMID- 12890315 TI - Global screening for human viral pathogens. AB - We propose a system for continuing surveillance of viral pathogens circulating in large human populations. We base this system on the physical isolation of viruses from large pooled samples of human serum and plasma (e.g., discarded specimens from diagnostic laboratories), followed by shotgun sequencing of the resulting genomes. The technology for concentrating virions from 100-L volumes was developed previously at Oak Ridge National Laboratory, and the means for purifying and concentrating virions from volumes in microliters have been developed recently. At the same time, marine virologists have developed efficient methods for concentrating, amplifying, and sequencing complex viral mixtures obtained from the ocean. Given this existing technology base, we believe an integrated, automated, and contained system for surveillance of the human "virome" can be implemented within 1 to 2 years. Such a system could monitor the levels of known viruses in human populations, rapidly detect outbreaks, and systematically discover novel or variant human viruses. PMID- 12890316 TI - Salmonella control programs in Denmark. AB - We describe Salmonella control programs of broiler chickens, layer hens, and pigs in Denmark. Major reductions in the incidence of foodborne human salmonellosis have occurred by integrated control of farms and food processing plants. Disease control has been achieved by monitoring the herds and flocks, eliminating infected animals, and diversifying animals (animals and products are processed differently depending on Salmonella status) and animal food products according to the determined risk. In 2001, the Danish society saved U.S.$25.5 million by controlling Salmonella. The total annual Salmonella control costs in year 2001 were U.S.$14.1 million (U.S.$0.075/kg of pork and U.S.$0.02/kg of broiler or egg). These costs are paid almost exclusively by the industry. The control principles described are applicable to most industrialized countries with modern intensive farming systems. PMID- 12890317 TI - Disease surveillance and the academic, clinical, and public health communities. AB - The Emerging Infections Programs (EIPs), a population-based network involving 10 state health departments and the Centers for Disease Control and Prevention, complement and support local, regional, and national surveillance and research efforts. EIPs depend on collaboration between public health agencies and clinical and academic institutions to perform active, population-based surveillance for infectious diseases; conduct applied epidemiologic and laboratory research; implement and evaluate pilot prevention and intervention projects; and provide capacity for flexible public health response. Recent EIP work has included monitoring the impact of a new conjugate vaccine on the epidemiology of invasive pneumococcal disease, providing the evidence base used to derive new recommendations to prevent neonatal group B streptococcal disease, measuring the impact of foodborne diseases in the United States, and developing a systematic, integrated laboratory and epidemiologic method for syndrome-based surveillance. PMID- 12890318 TI - Acute flaccid paralysis and West Nile virus infection. AB - Acute weakness associated with West Nile virus (WNV) infection has previously been attributed to a peripheral demyelinating process (Guillain-Barre syndrome); however, the exact etiology of this acute flaccid paralysis has not been systematically assessed. To thoroughly describe the clinical, laboratory, and electrodiagnostic features of this paralysis syndrome, we evaluated acute flaccid paralysis that developed in seven patients in the setting of acute WNV infection, consecutively identified in four hospitals in St. Tammany Parish and New Orleans, Louisiana, and Jackson, Mississippi. All patients had acute onset of asymmetric weakness and areflexia but no sensory abnormalities. Clinical and electrodiagnostic data suggested the involvement of spinal anterior horn cells, resulting in a poliomyelitis-like syndrome. In areas in which transmission is occurring, WNV infection should be considered in patients with acute flaccid paralysis. Recognition that such weakness may be of spinal origin may prevent inappropriate treatment and diagnostic testing. PMID- 12890319 TI - West Nile virus in farmed alligators. AB - Seven alligators were submitted to the Tifton Veterinary Diagnostic and Investigational Laboratory for necropsy during two epizootics in the fall of 2001 and 2002. The alligators were raised in temperature-controlled buildings and fed a diet of horsemeat supplemented with vitamins and minerals. Histologic findings in the juvenile alligators were multiorgan necrosis, heterophilic granulomas, and heterophilic perivasculitis and were most indicative of septicemia or bacteremia. Histologic findings in a hatchling alligator were random foci of necrosis in multiple organs and mononuclear perivascular encephalitis, indicative of a viral cause. West Nile virus was isolated from submissions in 2002. Reverse transcription-polymerase chain reaction (RT-PCR) results on all submitted case samples were positive for West Nile virus for one of four cases associated with the 2001 epizootic and three of three cases associated with the 2002 epizootic. RT-PCR analysis was positive for West Nile virus in the horsemeat collected during the 2002 outbreak but negative in the horsemeat collected after the outbreak. PMID- 12890320 TI - Molecular epidemiology of O139 Vibrio cholerae: mutation, lateral gene transfer, and founder flush. AB - Vibrio cholerae in O-group 139 was first isolated in 1992 and by 1993 had been found throughout the Indian subcontinent. This epidemic expansion probably resulted from a single source after a lateral gene transfer (LGT) event that changed the serotype of an epidemic V. cholerae O1 El Tor strain to O139. However, some studies found substantial genetic diversity, perhaps caused by multiple origins. To further explore the relatedness of O139 strains, we analyzed nine sequenced loci from 96 isolates from patients at the Infectious Diseases Hospital, Calcutta, from 1992 to 2000. We found 64 novel alleles distributed among 51 sequence types. LGT events produced three times the number of nucleotide changes compared to mutation. In contrast to the traditional concept of epidemic spread of a homogeneous clone, the establishment of variant alleles generated by LGT during the rapid expansion of a clonal bacterial population may be a paradigm in infections and epidemics. PMID- 12890321 TI - Amoeba-resisting bacteria and ventilator-associated pneumonia. AB - To evaluate the role of amoeba-associated bacteria as agents of ventilator associated pneumonia (VAP), we tested the water from an intensive care unit (ICU) every week for 6 months for such bacteria isolates; serum samples and bronchoalveolar lavage samples (BAL) were also obtained from 30 ICU patients. BAL samples were examined for amoeba-associated bacteria DNA by suicide-polymerase chain reaction, and serum samples were tested against ICU amoeba-associated bacteria. A total of 310 amoeba-associated bacteria from 10 species were isolated. Twelve of 30 serum samples seroconverted to one amoeba-associated bacterium isolated in the ICU, mainly Legionella anisa and Bosea massiliensis, the most common isolates from water (p=0.021). Amoeba-associated bacteria DNA was detected in BAL samples from two patients whose samples later seroconverted. Seroconversion was significantly associated with VAP and systemic inflammatory response syndrome, especially in patients for whom no etiologic agent was found by usual microbiologic investigations. Amoeba-associated bacteria might be a cause of VAP in ICUs, especially when microbiologic investigations are negative. PMID- 12890322 TI - Antimicrobial resistance markers of class 1 and class 2 integron-bearing Escherichia coli from irrigation water and sediments. AB - Municipal and agricultural pollution affects the Rio Grande, a river that separates the United States from Mexico. Three hundred and twenty-two Escherichia coli isolates were examined for multiple antibiotic resistance phenotypes and the prevalence of class 1 and class 2 integron sequences. Thirty-two (10%) of the isolates were resistant to multiple antibiotics. Four (13%) of these isolates contained class 1-specific integron sequences; one isolate contained class 2 integron-specific sequences. Sequencing showed that the class 1 integron-bearing strain contained two distinct gene cassettes, sat-1 and aadA. Although three of the four class 1 integron-bearing strains harbored the aadA sequence, none of the strains was phenotypically resistant to streptomycin. These results suggest that integron-bearing E. coli strains can be present in contaminated irrigation canals and that these isolates may not express these resistance markers. PMID- 12890323 TI - Hantavirus prevalence in the IX Region of Chile. AB - An epidemiologic and seroprevalence survey was conducted (n=830) to assess the proportion of persons exposed to hantavirus in IX Region Chile, which accounts for 25% of reported cases of hantavirus cardiopulmonary syndrome. This region has three geographic areas with different disease incidences and a high proportion of aboriginals. Serum samples were tested for immunoglobulin (Ig) G antibodies by enzyme-linked immunosorbent assay against Sin Nombre virus N antigen by strip immunoblot assay against Sin Nombre, Puumala, Rio Mamore, and Seoul N antigens. Samples from six patients were positive for IgG antibodies reactive with Andes virus; all patients lived in the Andes Mountains. Foresting was also associated with seropositivity; but not sex, age, race, rodent exposure, or farming activities. Exposure to hantavirus varies in different communities of IX Region. Absence of history of pneumonia or hospital admission in persons with specific IgG antibodies suggests that infection is clinically inapparent. PMID- 12890324 TI - Antimicrobial susceptibility breakpoints and first-step parC mutations in Streptococcus pneumoniae: redefining fluoroquinolone resistance. AB - Clinical antimicrobial susceptibility breakpoints are used to predict the clinical outcome of antimicrobial treatment. In contrast, microbiologic breakpoints are used to identify isolates that may be categorized as susceptible when applying clinical breakpoints but harbor resistance mechanisms that result in their reduced susceptibility to the agent being tested. Currently, the National Committee for Clinical Laboratory Standards (NCCLS) guidelines utilize clinical breakpoints to characterize the activity of the fluoroquinolones against Streptococcus pneumoniae. To determine whether levofloxacin breakpoints can identify isolates that harbor recognized resistance mechanisms, we examined 115 S. pneumoniae isolates with a levofloxacin MIC of >2 mg/mL for first-step parC mutations. A total of 48 (59%) of 82 isolates with a levofloxacin MIC of 2 mg/mL, a level considered susceptible by NCCLS criteria, had a first-step mutation in parC. Whether surveillance programs that use levofloxacin data can effectively detect emerging resistance and whether fluoroquinolones can effectively treat infections caused by such isolates should be evaluated. PMID- 12890325 TI - Mutations in putative mutator genes of Mycobacterium tuberculosis strains of the W-Beijing family. AB - Alterations in genes involved in the repair of DNA mutations (mut genes) result in an increased mutation frequency and better adaptability of the bacterium to stressful conditions. W-Beijing genotype strains displayed unique missense alterations in three putative mut genes, including two of the mutT type (Rv3908 and mutT2) and ogt. These polymorphisms were found to be characteristic and unique to W-Beijing phylogenetic lineage. Analysis of the mut genes in 55 representative W-Beijing isolates suggests a sequential acquisition of the mutations, elucidating a plausible pathway of the molecular evolution of this clonal family. The acquisition of mut genes may explain in part the ability of the isolates of W-Beijing type to rapidly adapt to their environment. PMID- 12890326 TI - Yellow pigmy rice rat (Oligoryzomys flavescens) and hantavirus pulmonary syndrome in Uruguay. AB - During 5,230 trapping nights, 672 small mammals were trapped in the areas where most hantavirus pulmonary syndrome (HPS) cases occur in Uruguay. Yellow pygmy rice rats (Oligoryzomys flavescens) were the only rodents that showed evidence of antibodies to hantavirus, with a seroprevalence of 2.6%. The rodents were trapped in all the explored environments, and most of the seropositive rodents were found in habitats frequented by humans. Nucleotide sequences were obtained from four HPS case-patients and four yellow pygmy rice rats of the M genome segment. Sequence comparison and phylogenetic analysis showed that rodent-borne viruses and viruses from three HPS case-patients form a well-supported clade and share a 96.4% identity with the previously characterized Central Plata hantavirus. These results suggest that yellow pygmy rice rat (O. flavescens) may be the host for Central Plata, a hantavirus associated with HPS in the southern area of Uruguay. PMID- 12890327 TI - Serologic evidence of West Nile virus infection in horses, Coahuila State, Mexico. AB - Serum samples were obtained from 24 horses in the State of Coahuila, Mexico, in December 2002. Antibodies to West Nile virus were detected by epitope-blocking enzyme-linked immunosorbent assay and confirmed by plaque reduction neutralization test in 15 (62.5%) horses. We report the first West Nile virus activity in northern Mexico. PMID- 12890328 TI - Serologic evidence of West Nile virus infection in horses, Yucatan State, Mexico. AB - Serum samples were obtained from 252 horses in the State of Yucatan, Mexico, from July to October 2002. Antibodies to West Nile virus were detected by epitope blocking enzyme-linked immunosorbent assays in three (1.2%) horses and confirmed by plaque reduction neutralization test. We report the first West Nile virus activity in the State of Yucatan. PMID- 12890329 TI - Serologic evidence of West Nile virus transmission, Jamaica, West Indies. AB - In spring 2002, an intensive avian serosurvey was initiated in Jamaica, Puerto Rico, and Mexico. We collected >1,600 specimens from resident and nonresident neotropical migratory birds before their northerly migrations. Plaque reduction neutralization test results indicated specific neutralizing antibodies to West Nile virus in 11 resident species from Jamaica. PMID- 12890330 TI - Sulfa resistance and dihydropteroate synthase mutants in recurrent Pneumocystis carinii pneumonia. AB - Failure of sulfa or sulfone prophylaxis is associated with mutations in Pneumocystis carinii gene coding for dihydropteroate synthase (DHPS). The DHPS genotype was analyzed in AIDS patients who had two separate episodes of P. carinii pneumonia. The results suggest that DHPS mutations can be selected de novo within patients by the pressure of a sulfa or sulfone drug. PMID- 12890331 TI - VIM- and IMP-type metallo-beta-lactamase-producing Pseudomonas spp. and Acinetobacter spp. in Korean hospitals. AB - We determined the occurrence of acquired metallo-b-lactamase (MBL)-producing bacteria in Korean hospitals. Among the isolates nonsusceptible to imipenem that were collected from 28 hospitals from 2000 to 2001, 44 (11.4%) of 387 Pseudomonas spp. and 38 (14.2%) of 267 Acinetobacter spp. produced MBL and had alleles of blaVIM-2 or blaIMP-1. MBL-producing isolates were detected in 60.7% of the hospitals. PMID- 12890332 TI - Leishmaniasis in Germany. AB - In 2000, a reference center was created to systematically record leishmaniases in Germany. We analyzed 58 cases of leishmaniases imported during a 2-year period. These findings will serve as a baseline for the sandfly vector's anticipated northward move because of global warming and as an advisory for immunocompromised persons traveling to leishmaniasis-endemic areas. PMID- 12890333 TI - Probable dengue virus infection among Italian troops, East Timor, 1999-2000. AB - To investigate the attack rate and risk factors for probable dengue fever, a cross-sectional study was conducted of an Italian military unit after its deployment to East Timor. Probable dengue was contracted by 16 (6.6%) of 241 army troops and caused half of all medical evacuations (12/24); no cases were detected among navy and air force personnel. PMID- 12890334 TI - HIV epidemic among young Thai Men, 1991-2000. AB - Characterization of the HIV epidemic in Thailand has benefited from the systematic testing of young men upon entry into the military. These data, which have shown that public health measures can reverse an HIV epidemic, have been reanalyzed with current geographic information systems methods. The resulting maps are, thus far, the best means of visualizing the geography of the dynamic HIV epidemic in Thailand. PMID- 12890335 TI - [Consensus and progresses of spontaneous bacterial peritonitis]. PMID- 12890336 TI - [Clinical analysis of drug-induced liver disease in 191 patients]. PMID- 12890337 TI - [Autologous adoptive immunotherapy without destruction of infected cells during the treatment of chronic hepatitis B]. AB - OBJECTIVES: To investigate the profile of liver histological damage after autologous adoptive immunotherapy with cytokine-induced killer cells (CIK cells) in patients with chronic hepatitis B (CHB). METHODS: 16 CHB patients were randomly enrolled and received autologous adoptive immunotherapy, then followed up 52 weeks. Liver samples were taken from the patients to evaluate the degree of inflammation and fibrosis. The markers of hepatitis B virus and liver function were also detected. RESULTS: 4 patients had two liver-biopsied samples before therapy and after 52 weeks follow-up. One patient's histological assessment revealed a significant improvement in intralobular necroinflammation (G2 --> G1) and fibrosis (S2 --> S1). The others failed to show obvious changes in liver histology. After 10 days culture in vitro, phenotypic characterization of CIK cells changed significantly by flow cytometry. The percentage of CD4+ cells decreased gradually, while the percentage of CD8+ cells increased from 20% to 60% - 80%. After 52 weeks follow-up, HBV DNA was negative (HBV DNA<4pg/ml in serum) in 6 out of 14 patients. The rates of both HBeAg/anti-HBe seroconversion and alanine aminotransferase normalization were 42.86%(6/14). There was no HBsAg/anti HBs seroconversion. There was few severe treatment-related adverse events. CONCLUSION: Autologous adoptive immunotherapy doesn't induce the damage of liver histology in chronic hepatitis B patients, which inhibits hepatitis B virus replication by a certain noncytotoxic mechanism. PMID- 12890338 TI - [Detection of hepatic progenitor cells in patients with severe hepatitis and their distribution]. AB - OBJECTIVES: To identify hepatic progenitor cells (HPCs) in patients with severe hepatitis (SH) by detecting their markers and investigate the features of their distribution and location. METHODS: Liver tissues taken from 59 SH patients were tested for the receptor of stem cell factor (c-kit), pi-class glutathione S transferase (GST-pi), cluster of differentiation 34 (CD34), cytokeratin 19 (CK19), cytokeratin 18 (CK18) and alpha fetoprotein (AFP) by immunohistochemistry (IHC). Meanwhile, 58 patients with acute or chronic hepatitis were also detected to act as controls. RESULTS: Hepatic progenitor cells could be seen in SH patients. Most of them existed as ductular cells that had been called "typical ductular proliferation (ADP)" or "typical ductular reaction" in previous research. These ductular cells were mainly located at the portal areas, fibro septa, periportal parenchyma and the border of the pseudolobuli and inflammatory foci. Further, c-kit, GST-pi, CK19 and CK18, but not CD34 and AFP could be detected in these cells. Another kind of HPC was the small hepatocyte-like cell (SHLC), which could express c-kit, GST-pi, and CK18, but not CK19, CD34 and AFP. The semi-quantitative analysis showed that the scope of ADP in SH patients was significantly larger than that in acute and chronic hepatitis patients (chi2= 63.62, P<0.05), and the scope of ADP in subacute severe hepatitis and chronic severe hepatitis patients was also significantly larger than that in acute severe hepatitis patients. CONCLUSION: In the course of regeneration of viral hepatitis, different types of pathology have different features. In acute and chronic hepatitis (G1-2), the regeneration is mainly owing to the proliferation of mature hepatocytes, and in chronic hepatitis (G3-4), there is the participation of HPCs, although they are limited. In severe hepatitis, however, since the replicative capacity of normal hepatocytes is impaired or prohibited, liver regenerates and restores mainly by the means of hepatic stem cells activation and proliferation. But the hepatic stem cells don't differentiate into their mature functional compartments directly at all. There are several intermediary or transition populations. In human severe hepatitis, they are mainly ductular cells, and parts of them are small hepatocyte-like cells. PMID- 12890339 TI - [PreS/S gene mutations in HBV in children infected through mother-to-infant transmission and in their mothers]. AB - OBJECTIVES: To investigate the characteristics of mutations in PreS/S gene of HBV in children infected through mother-to-infant transmission and in their mothers with different degree of viremia. METHODS: There were 15 pairs of child and mother in this study. Mothers of all children were chronic asymptomatic HBsAg carrier (ASC) before pregnancy and the children were not inoculated against HBV after birth. Anti-HBV medicine was never administrated to all subjects. The serological markers of hepatitis A, B, C, D and E virus were tested and the titers of serum HBV DNA were quantitated. PreS/S gene was amplified by PCR and cloned into pGEM-T vector with T-A cloning technique. The recombinant plasmid pGEM-PreS/S was confirmed by digestion with restriction enzyme ApaI and SacI. Two clones were selected to be sequenced from each patient. RESULTS: According to the degree of viremia in every pair of mother and child, 15 pairs of child and mother were divided into three groups: group A (both children and mothers had high viremia with HBeAg-positive), group B (high in children and low in mothers with anti-HBe positive), and group C (low in children and high in mothers), and there were 5 pairs in each group. The subtype of each pair was the same. There were 4/5 pairs of HBV with B/adw2 and 1/5 pair of HBV with C/adrq+ in each group. It was shown that there were no difference among the four high viremia groups or between the two low viremia groups in the number of mutations and the number of mutational positions. However, there was significant difference between high viremia group and low viremia group. The mutation was not related to age. There were 56 mutational positions and there was no mutational hotspot in high viremia patients. In two low viremia groups (the mothers in group B and the children in group C), there were 113 mutational positions and 85 mutational positions were hotspots (owned by 5/8 clones in each) which could make 37 amino acids changed. Most of mutational amino acids were located within T and B cell epitopes of envelope protein or/and located in the surrounding regions. CONCLUSIONS: There are many differences in HBV with different degree of viremia, even if it comes from the same strain. There are some regular patterns in the mutations of HBV after HBeAg seroconversion happened. PMID- 12890340 TI - [Preparation and application of the national reference panel for hepatitis B virus DNA]. AB - OBJECTIVES: To calibrate the national hepatitis B virus (HBV) DNA standard according to world health organization's standard material and prepare the national reference panel for HBV DNA reagents. METHODS: Sera from blood donors and HBV patients were collected and detected by home-made HBV DNA PCR kits, HBsAg kits, and anti-HBc kits, and then confirmed by HBV DNA PCR kits produced by Roche in German, which was recognized by the world health organization. The stability of the panel was detected by acceleration method. RESULTS: The convinced copies of the sensitivity samples were gotten by seven independent experiments, the coefficients of variation of logarithm of the copies of L0-L5 were all less than 15%. Regarding the national reference panel as the standard, the quality of most domestic HBV DNA PCR kits was improved, while part of the kits should be further qualified. CONCLUSION: The national reference panel for HBV DNA reagents is developed. It contains eight negative, nine positive sera and seven samples for sensitivity test PMID- 12890341 TI - [Detection of anti-HEV IgG and analysis of partial HEV RNA sequence]. AB - OBJECTIVES: To investigate HEV infection in swine and the genotype relationship between swine and human HEV. METHODS: Anti-HEV IgG antibody was detected in the sera of swine using enzyme linked immunoassay (EIA), and HEV RNA was amplified by reverse transcription nested polymerase chain reaction (RT-nPCR). The Vector NTI Suite 7 and TreeView softwares were used for nucleotide sequences phylogenetic analysis of HEV isolated from human and swine. RESULTS: The anti-HEV IgG positive rate was 16.67% (18/108). Among the 18 anti-HEV IgG positive sera, 2 sequences (11.11%, called S18 and S43, respectively) of HEV ORF1 (102-387bp) were amplified, with the identity of 99% between them. They had 76% to 77%, 78%, 76% to 79%, 85% to 86%, 77%, 80%, 79% and 75% - 79% homology at the nucleotide level with human HEV genotypes 1 to 8, respectively. One (S18) of them was also amplified out in ORF2 region (5,994-6 297bp) and showed 76% to 78%, 74%, 74% to 77%, and 85% to 94% identity with human HEV genotypes 1 to 4 at the nucleotide level, respectively. CONCLUSION: HEV sequences isolated from swine belong to human HEV genotype 4. PMID- 12890342 TI - [Therapeutic effects of octreotide on hepatofibrosis-induced with carbon tetrachloride in rats]. AB - OBJECTIVES: To investigate the therapeutic effects and mechanism of octreotide on experimental hepatic fibrosis in rats. METHODS: Hepatofibrotic rats models were established with carbon tetrachloride. All the experimental rats were divided into four groups: normal control group, pre-and post-treatment model group, and octreotide-treated group in which the rats were injected subcutaneously with octreotide at the dose of 50ng/100g, twice daily, for thirty days. Serum levels of hyaluronic acid (HA), laminin (LN) and pro-collagen type III peptide (PCIII) were detected by radioimmunoassay. Hepatic fibrosis scoring grade was assessed through Van-Gieson staining and observed under light microscope. Protein expression levels of alpha-smooth muscle actin (alpha-SMA) and transforming growth factor beta1 (TGFbeta1) were determined with immunohistochemical staining method. Messenger RNA (mRNA) levels of collagen type I and PCIII were detected by reverse transcription polymerase chain reaction. RESULTS: Serum levels of HA (ng/L), LN (microg/L) and PCIII (ng/L) in pre- and post-treatment model groups were higher than those in normal control group (121.8+/-9.5 and 110.3+/-13.4 vs. 33.1+/-3.7, 85.7+/-12.1 and 78.2+/-7.9 vs. 37.1+/-6.3, 35.9+/-3.5 and 33.7+/-2.6 vs. 15.6+/-2.8, respectively, t > or = 9.41, P<0.05), and there was no significant difference between the two model groups. Concentrations of HA (55.8ng/L+/-7.2ng/L), LN (43.1microg/L+/-3.4microg/L) and PCIII (27.8ng/L+/ 3.4ng/L) decreased significantly in octreotide-treated group, compared with those in model groups (t >or=2.76, P<0.05). With histological analysis, fibrotic scoring grade in octreotide-treated group was obviously ameliorated, compared with that in model groups (chi2 > or = 3.97, P<0.05). Imaging analysis revealed that alpha-SMA and TGFbeta1 immunohistological staining areas were markedly shrinked in octreotide-treated group (t > or = 2.47, P < 0.05). In two model groups, PCIII and type I mRNA levels significantly up-regulated as compared with those in normal group (t > or = 9.27, P<0.001), and they were inhibited by octreotide markedly (t > or = 2.47, P<0.05). CONCLUSIONS: Octreotide can inhibit hepatic stellate cells transforming into myofibroblasts, down-regulate TGFbeta1, collagen type I and PCIII transcriptions, so that it has therapeutic effects on experimental hepatic fibrosis. PMID- 12890343 TI - [Detection of platelet Ca2+(i), CD62P, CD63 and plasma CD62P in cirrhosis patients]. AB - OBJECTIVES: To study on the relationship between platelet Ca2+(i), CD62P, CD63, serum CD62P (SCD62P) and cirrhosis patients. METHODS: Platelet CD62P, CD63 were determined with flow cytometry, SCD63P with ELISA, and Ca2+(i) in platelet was determined with fluorophotometry. RESULTS: Platelet Ca2+(i), CD62P, CD63, and SCD62P levels in cirrhosis patients were (103.1+/-22.2)nmol/L, (47.6+/-20.0)%, (47.1+/-24.6)%, and (67.6+/-37.6)microg/L, and in controls were (57.6+/ 13.1)nmol/L, (3.1+/-0.7)%, (2.5+/-0.7)%, and (24.0+/-6.5)microg/L, respectively. The levels in the former were higher than those in the latter (t > or = 6.148, P<0.05). The above levels in upper gastrointestinal haemorrhage group were much higher than those in the non-haemorrhage group (120.3nmol/L+/-18.8nmol/L vs 91.1nmol/L+/-14.3nmol/L, 64.9%+/-14.7% vs 34.6%+/- 11.9%, 70.9%+/-14.5% vs 30.2%+/-14.4%, and 103.6microg/L+/-14.9microg/L vs 40.8microg/L+/-24.0microg/L, respectively, t > or = 5.380, P<0.05). But the numbers of platelet between the two groups were no obvious difference. CONCLUSIONS: Platelet in the cirrhosis patients is greatly active, and the detection of platelet CD62P, CD63, SCD62P has a certain value in judging the degree of cirrhosis. PMID- 12890344 TI - [Initial research of the effect and mechanism of cytotoxic T lymphocyte (lymphocyte) primed by dendritic cell as hepatocellular carcinoma vaccine]. PMID- 12890345 TI - [Effects of PreS2 synthetic peptides on infiltrating T lymphocyte subgroups expression in hepatocellular carcinoma]. AB - OBJECTIVES: To explore whether PreS2 can change the percentage of T lymphocyte subgroups and the ration of CD4+/CD8+ in hepatocellular carcinoma (HCC) caused by HBV. METHODS: The P120-146 region composed by the way of Merrifield, which was the most intensive antigen in PreS2 peptides, served as the antigen after dissolved in 0.01 mol/L PBS. 12 patients were chosed as the subjects, who were pathologically diagnosed as HCC after operation, were HBsAg-, HBeAg-, anti-HBc, and HBV DNA positive in serum, and expressed HBsAg in HCC tissue. The monocytes were isolated and cultured in 96 microplate with 1x 10(6) cells in every well, then the PreS2 synthetic peptides was added in at the doses of 1microg, 5microg, and 10microg, also IL-2 with 500 U was added in. Seven days later, the percentage of CD3+, CD4+, CD8+, and the ratio of CD4+/CD8+ were detected. RESULTS: It was found that the percentage of CD4+ increased significantly (t = 3.508, P < 0.01), and the ratio of CD4+/CD8+ decreasedly obviously (t = 2.235, P < 0.05) in the 5microg PreS2 synthetic peptides group, compared with those in the control group. The percentage of CD3+ rised markedly in the IL-2 group, compared with that in the control group. CONCLUSION: With proper doses, PreS2 is capable of changing the expression of T lymphocyte subgroups in HCC tissue, increasing the percentage of CD4+ obviously and changing the motionless state of CD8+, to make the carcinoma cell killed through the action of CD4+ and CD8+. PMID- 12890346 TI - [Changes of liver function in patients with serious acute respiratory syndrome]. AB - OBJECTIVES: To investigate the changes of liver function during the course of serious acute respiratory syndrome (SARS) and to explore its possible influence factors. METHODS: There were 91 patients with common SARS, and 23 patients with severe SARS, and 61 common pneumonia patients served as the controls. The liver functions of all the patients were measured. RESULTS: The rate of anomaly liver function in the common SARS patients group was 68.1%, which was higher than that in the common pneumonia patients group (24.6%), chi2=27.7, P<0.01. The changes mainly existed in the mild to moderate elevation of alanine aminotransferase and aspartate aminotransferase. The severe SARS patients were older and the changing rate of liver function was as high as 95.7%. CONCLUSIONS: The damage possibility of liver function in SARS patients is higher than that in common pneumonia patients. The damage is light and related to SARS itself. The damage of liver function in the severe SARS patients may have close relationship with age. PMID- 12890347 TI - [Transfering into accessory nasal sinuses and bone in a primary hepatocellular carcinoma patient]. PMID- 12890348 TI - [Expression of rat augmenter of liver regeneration in pichia pastoris and evaluation of its bioactivity in vitro]. AB - OBJECTIVES: To construct yeast expression plasmid of rat augmenter of liver regeneration (rALR), express it in GS115, and examine its bioactivity in vitro. METHODS: With PCR and genetic recombination techniques, the gene fragment of rALR was amplified from recombinant plasmid pcDNA3.1-rALR, and the recombinant plasmid pPIC9K-rALR was constructed. The recombinant plasmid pPIC9K-rALR was transducted into GS115 by electroporation after identified with endonuclease digestion and PCR amplification. The target protein was expressed by GS115 under the induction of 0.5% methanol. The recombinant rALR (rrALR) was purified with ultrafiltration after demonstrated by 15% SDS-PAGE and western blot. The effects of rrALR on the proliferation of QGY, HepG2 cells and primary rat hepatocytes in vitro were evaluated by 3H-TdR intake method. RESULTS: Recombinant plasmid pPIC9K-rALR was identified by restriction digestion and PCR methods. The rrALR as a secretive protein was successfully expressed in GS115. Its molecular weight, 1.5x10(4), was in correspondence with theoretic value. The rrALR could bind with the polyclonal antibody against human ALR in western blot, which demonstrated that there was cross-antigenicity between human and rat ALR. The high qualitative rrALR was obtained through ultrafiltration. Among the experimental concentrations, the rrALR could stimulate the proliferations of QGY and HepG2 cells in a dose dependent manner in vitro, but had little effect on the proliferation of primary rat hepatocytes in vitro. CONCLUSIONS: The rrALR as a secretive protein is expressed in GS115 efficiently. The rrALR can stimulate the proliferations of QGY and HepG2 cells in a dose-dependent manner in vitro, but had no effect on primary rat hepatocytes, which demonstrates that there exists different receptors on the normal hepatocytes and hepatocarcinoma cells. There are cross-bioactivity and cross-antigenicity between human and rat ALR. PMID- 12890349 TI - [TLR4 is involved in hepatic ischemia/reperfusion injury in mice]. AB - OBJECTIVES: To explore the role of TLR4 in the mechanism of hepatic ischemia/reperfusion (I/R) injury in mice. METHODS: Wild-type (C3H/Heouj) mice and TLR4 deficient mice (C3H/Hej) were used to prepare the models of liver I/R injury. Partial hepatic ischemia was produced by inflow causing occlusion in the median and left lobes for 45 minutes. Blood was drawn to kill the mice at 1 hours and 3 hours after reperfusion. The blood was used to analyze aspartate aminotransferase (AST) and tumor necrosis factor-alpha (TNFalpha). TNF-alpha mRNA expression and myeloperoxidase (MPO) level in ischemic lobes was examined by northern blot and myeloperoxidase assay, respectively. RESULTS: AST levels were significantly lower in TLR4 deficient mice, compared with those in wild-type mice at both time points (661.83U/L+/-106.09U/L vs. 1215.5U/L+/- 174.03U/L, t=-6.65, P<0.01; 1145.17U/L+/-132.42U/L vs. 2958.17U/L+/-186.81U/L, t=-5.57, P<0.01). Serum TNF-alpha level was lower in TLR4 deficient mice at 3 hours after reperfusion compared with that in wild-type mice (152.39pg/ml+/-43.3 pg/ml vs. 249.12pg/ml+/-51.89pg/ml, t=-3.13, P<0.05). This difference appeared to be mediated at the gene level, since TNF-alpha mRNA expression had decreased in TLR4 deficient mice at 1 hours after reperfusion, compared with that in wild type mice (80.3+/-28.8 vs. 189.4+/-24.6, t=-3.25, P<0.05). MPO level in ischemic lobes in TLR4 deficient mice at 3 hours after reperfusion was significantly lower than that in wild type mice (F=33.49, P<0.01). CONCLUSIONS: I/R hepatic injury in TLR4 deficient mice is less than that in wild-type mice. TNF-alpha expression down regulated at the mRNA level appears critical. These suggest that TLR4 be involved in the mechanism of hepatic ischemia/reperfusion injury in mice. PMID- 12890350 TI - [Evaluation of different methods in monitoring YMDD motif mutations associated with lamivudine resistance]. AB - OBJECTIVES: To evaluate the three different methods in monitoring the lamivudine resistant HBV mutants in lamivudine-treated patients with chronic hepatitis B. METHODS: The sensitivity and specialty of melting curve assay and polymerase chain reaction microplate nucleotide hybridization-enzyme linked immunosorbent assay (PCRmnh-ELISA) were compared with those of mismatch polymerase chain reaction-restriction fragment length polymorphism (mPCR-RFLP) and sequence analysis, through detection of HBV YMDD mutants in 44 serums from chronic hepatitis B patients receiving lamivudine monotherapy at the time of viral breakthrough. RESULTS: mPCR-RFLP assay was more sensitive (10(4) copies/ml) than both PCRmnh-ELISA (10(5) copies/ml) and melting curve assay (10(6) copies/ml). 26 YMDD mutants and 18 wild-types were determined by the means of mPCR-RFLP. Among the 26 mutants, only 16 and 18 mutants were found by melting curve assay and PCRmnh-ELISA, respectively. Whereas, out of the 18 wild-types, 2 and 13 mutants were detected by melting curve assay and PCRmnh-ELISA, respectively. To confirm the different results determined by the three methods in 16 samples, sequence analysis was conducted and showed that the rate of consistency with sequencing was 93.8% by mPCR-RFLP, 43.8% by melting curve, and 18.8% by PCRmnh-ELISA, respectively (chi2=18.7, P<0.01). CONCLUSIONS: The mPCR-RFLP assay is reliable to monitor HBV YMDD mutations. Melting curve assay and PCRmnh-ELISA should be further improved to increase their sensitivity and specialty. PMID- 12890351 TI - [Detection of YMDD motif mutations in lamivudine-untreated patients with chronic hepatitis B]. PMID- 12890352 TI - [Construction of the expression vectors of HDV ribozymes and their intracellular inhibiting activity against HCV RNA]. AB - OBJECTIVES: To investigate whether HDV ribozymes can intracellularly inhibit HCV RNA. METHODS: The mammalian expression vectors, pC1-RzC1, pC1-RzC2 and pC1-RzC3, containing ribozymes cDNA of RzC1, RzC2, and RzC3, were constructed targeting different HCV-5' NCR-C RNA regions. Then the HCV-positive fetal hepatocytes were transfected with these plasmids using liposome-mediated method. The inhibitory effects of HDV ribozymes were evaluated by HCV RNA quantitation in cultured cells and the supernatants. RESULTS: (1) All the three HDV ribozymes were inserted into the expression vector. (2) Fetal hepatocytes were infected with HCV proven by RT PCR and fluorescent quantitative PCR and expressed HCV NS3 and NS5 antigens by immunocytochemistry. (3) HDV ribozymes inhibited the activity of the target HCV RNA at expect positions in HCV-positive hepatocytes. At 0.5 micromol/L, the inhibitory rate of pC1-RzC1, pC1-RzC2, and pC1-RzC3 was 53.2%, 50.5 %, and 10.6% respectively. PC1-RzC1 was used continuously for one week, showing the inhibitory rate of 60.7%, 64.2%, 68.4%, 71.9%, 78.8% and 83.1% on the 2nd, 3rd, 4th, 5th, 6th and 7th day. CONCLUSION: The inhibitory activity of pC1-RzC1 (107-113nt) and pC1-RzC2 (268-274nt) is greater than that of pC1-RzC3 (345-351nt) in HCV-positive hepatocytes. PMID- 12890353 TI - [The factors leading hepatitis C to chronicnization]. PMID- 12890354 TI - [Influence of hot spot mutations in HBV/C gene on T cell epitopes with HLA-A2]. PMID- 12890355 TI - [Spontaneous bacterial peritonitis]. PMID- 12890356 TI - [Effects of regulatory endoplasmic HBX expression on hepatic cell apoptosis]. PMID- 12890357 TI - [A clinical analysis of spontaneous bacterial peritonitis in patients with severe hepatitis and hepatic cirrhosis]. PMID- 12890358 TI - [Study the mutations in polymerase gene of hepatitis B virus by three methods]. PMID- 12890359 TI - [Immunogenicity of a DNA vaccine coexpressing hepatitis B virus surface-core fusion antigen and IL-12]. PMID- 12890360 TI - [Immunohistochemical study of ductular proliferation in patients with chronic severe hepatitis]. PMID- 12890361 TI - [Inhibition of the replication of hepatitis B virus in vitro by a novel nucleoside analogue (beta-L-D4A)]. PMID- 12890362 TI - [Renin angiotensin system and liver fibrosis or cirrhosis]. PMID- 12890363 TI - The chest X-ray image features of patients with severe SRAS: a preliminary study. AB - OBJECTIVE: To study the chest X-ray image features of patients with severe SARS. METHODS: Chest X-ray image features in 36 patients with severe SARS were retrospectively analyzed. The image characteristics were compared with those of 224 patients with common SARS. RESULTS: The important chest X-ray imaging features of 36 patients with severe SARS included small patch of infiltration (n = 27, 75.0%), large patch of infiltration (n = 22, 61.1%), large area of lung consolidation (n = 10, 27.3%), interstitial lung lesion (n = 26, 72.2%), ground glass shadow (n = 28, 77.8%), irregular linear opacity (n = 15, 41.7%), diffuse lung lesion (n = 12, 33.3%), with single lung involved (n = 9, 25.0%), and both lungs involved (n = 32, 88.9%). The rates of large patch of infiltration, large area of lung consolidation, ground-glass shadow, diffuse lung lesion and involvement of both lungs in patients with severe SARS were significantly higher than those in patients with common type of SARS (all P < 0.01). Out of the 11 severe SARS patients who died, nine had large area of ground-glass shadow with air bronchogram in both lungs before death. CONCLUSIONS: Large patch of infiltration, large area of consolidation, ground-glass shadow, diffuse lung lesion and involvement of both lungs were the main X-ray image characteristics of patients with severe SARS. Large area of ground-glass shadow with air bronchogram in both lungs indicated a bad prognosis. PMID- 12890364 TI - Chest X-ray imaging of patients with SARS. AB - OBJECTIVE: To investigate the chest X-ray manifestations of SARS cases. METHODS: A retrospective study was conducted among 52 clinically confirmed SARS patients from February 9 to May 10, 2003. Chest X-ray scanning was performed at a interval of 1 - 3 days according to the requirements. The manifestations and special features of SARS in X-ray were analyzed. RESULTS: Small or large patchy shadows with intensive density in both lungs were observed in 31 cases, ground-glass like opacification in 16, small patchy shadows in one lung lobe or one lung segment in 18, nodular shadows in one lung segment in 1, and increased lung marking in lung interstitial tissues in 2. Rapidly changing consolidations revealed in chest X ray images were found to be associated with SARS infections, and they were not affected by treatment with antibiotics. CONCLUSION: Chest X-ray provides a sensitive and specific method for the diagnosis and treatment of SARS, and those present with symptoms and signs should undergo chest X-ray scanning every 1 - 3 days. PMID- 12890365 TI - Pathological study on severe acute respiratory syndrome. AB - OBJECTIVE: To study the pathological characteristics of severe acute respiratory syndrome (SARS) and its relationship to clinical manifestation. METHODS: Tissue specimens from 3 autopsies of probable SARS cases were studied by microscope, and the clinical data was reviewed. RESULTS: The typical pathological changes of lungs were diffuse hemorrhaging on the surface. A combination of serous, fibrinous and hemorrhagic inflammation was seen in most of the pulmonary alveoli with the engorgement of capillaries and detection of micro-thrombosis in some of these capillaries. Pulmonary alveoli thickened with interstitial mononuclear inflammatory infiltrates, suffered diffuse alveolar damage, experienced desquamation of pneumocytes and had hyaline-membrane formation, fibrinoid materials, and erythrocytes in alveolar spaces. There were thromboembolisms in some bronchial arteries. Furthermore, hemorrhagic necrosis was also evident in lymph nodes and spleen with the attenuation of lymphocytes. Other atypical pathological changes, such as hydropic degeneration, fatty degeneration, interstitial cell proliferation and lesions having existed before hospitalization were observed in the liver, heart, kidney and pancreas. CONCLUSION: Severe damage to the pulmonary and immunological systems is responsible for the clinical features of SARS and may lead to the death of patients. PMID- 12890366 TI - The relationship between serum interleukins and T-lymphocyte subsets in patients with severe acute respiratory syndrome. AB - OBJECTIVES: To observe the changes of serum interleukins (IL), T-lymphocyte subsets, and white blood cell (WBC) count in patients with severe acute respiratory syndrome (SARS), and to investigate the relationship between injured immune function, immune response and disturbed immune adjustment in SARS patients. METHODS: The levels of serum IL-2, IL-10, IL-12 and T-lymphocyte subset counts were measured in 35 clinically diagnosed SARS patients by using enzyme linked immunosorbant assay (ELISA). The relationship between the measured results and WBC count was further analyzed. RESULTS: The level of serum IL was increased to a great extent in the 35 SARS patients, and the levels of serum IL-2, IL-10 and IL-12 were 242.53 (92.69) pg/ml, 77.43 (63.37) pg/ml and 65.94 (43.21) pg/ml, respectively. The level of serum IL-2 increased markedly (P < 0.01). The peripheral blood CD(3)(+), CD(4)(+) and CD(8)(+) counts were lower than normal in 23 patients (67.7%), 26 patients (74.3%) and 15 patients (42.9%), respectively. The peripheral blood WBC counts were lower than 4.0 x 10(9)/L in 10 patients, and their CD(3)(+), CD(4)(+) and CD(8)(+) counts were 583.90 (315.58) x 10(6)/L, 272.00 (94.13) x 10(6)/L and 209.00 (72.21) x 10(6)/L, respectively. The peripheral blood WBC counts were (4.0 - 10.0) x 10(9)/L in 20 patients, and their CD(3)(+), CD(4)(+) and CD(8)(+) counts were 700.00 (502.96) x 10(6)/L, 347.00 (247.58) x 10(6)/L and 322.05 (228.47) x 10(6)/L, respectively. The peripheral blood WBC counts were higher than 10.0 x 10(9)/L in 5 patients, and their CD(3)(+), CD(4)(+) and CD(8)(+) counts were 1466.00 (630.86) x 10(6)/L, 783.00 (311.14) x 10(6)/L and 640.00 (294.40) x 10(6)/L, respectively. The decreased CD(3)(+), CD(4)(+) and CD(8)(+) counts were consistent with the decreased WBC counts. The level of IL in SARS patients was significantly higher than that in patients with chronic hepatitis B (P < 0.01). CONCLUSIONS: The level of serum IL is closely related to cell immunity in SARS patients. The level of serum IL is increased evidently while CD(3)(+), CD(4)(+) and CD(8)(+) counts decrease. Both serum IL and CD are associated with injury of immune function, and thus they could be regarded as a monitoring index for judging the condition of SARS patients and prescribing immune therapy. PMID- 12890367 TI - Rapid loss of both CD4+ and CD8+ T lymphocyte subsets during the acute phase of severe acute respiratory syndrome. AB - OBJECTIVE: To study the alteration of peripheral lymphocyte subsets in severe acute respiratory syndrome (SARS) patients and to help improve the early diagnosis of the disease. METHODS: Anti-coagulating blood samples from 98 SARS patients in the acute phase, 56 normal healthy blood donors, and from patients infected by HIV, CMV and EBV were collected. The T lymphocyte subsets were counted by flow cytometry using fluorescence-labeled specific monoclonal antibodies. RESULTS: A significant decrease was observed in all SARS patients in their peripheral CD4(+) and CD8(+) T lymphocyte absolute counts [256 (104) x 10(6)/L and 256 (117) x 10(6)/L, respectively], which were also lower than those of the patients infected with HIV, CMV and EBV. All patients infected with HIV, CMV and EBV had significantly higher CD8(+) T lymphocyte counts in comparison with normal controls. CONCLUSIONS: Decrease of both CD4(+) and CD8(+) T lymphocytes of patients is related to onset of SARS. T lymphocyte subset analysis would help improve the early diagnosis of the disease. PMID- 12890368 TI - Establishment of a fluorescent polymerase chain reaction method for the detection of the SARS-associated coronavirus and its clinical application. AB - OBJECTIVE: To establish a fluorescent polymerase chain reaction (F-PCR) method for detecting the coronavirus related to severe acute respiratory syndrome (SARS) and to evaluate its value for clinical application. METHODS: The primers and the fluorescence-labeled probe were designed and synthesized according to the published sequence of the SARS-associated coronavirus genes. A F-PCR diagnosis kit for detecting the coronavirus was developed, and 115 clinical nasopharyngeal gargling liquid samples were tested. RESULTS: The sequence of PCR amplified products completely matched the related sequence of the SARS-associated coronavirus genome. Forty-nine out of 67 samples from identified SARS patients and 8 of 18 samples from persons having close contact with SARS patients showed positive results. All 30 samples from healthy controls were negative. CONCLUSION: The F-PCR method established may be a rapid, accurate and efficient way for screening and for the early diagnosis of SARS patients. PMID- 12890369 TI - Correlation of pulmonary functions of COPD patients to those of their first degree children. AB - OBJECTIVE: To assess the risk factors correlating to the likelihood for airflow obstruction among first-degree children of chronic obstructive pulmonary disease (COPD) patients and whether familial aggregation of pulmonary function abnormality exists. METHODS: Fifty-nine smokers with COPD and 28 smokers without COPD as control and all their children available were recruited into the study. Their history was recorded and a binary logistic regression analysis was carried out to ascertain the effects of their relationship to a proband with COPD, when other potential risk factors were controlled. RESULTS: Children with COPD probands showed increased risk of FEV1 below the 70% predicted (OR = 1.987) after accounting for the effects of smoking, sex and clinical symptoms. The lower the pulmonary function of the COPD proband, the higher the risk to their children for FEV1 below the 70% predicted. CONCLUSIONS: Our finding demonstrates the presence of a household aggregation inclination of COPD and pulmonary function impairment. Genetic factors might act as the basis of the familial aggregation. PMID- 12890370 TI - Effect of volume replacement with hydroxyethyl starch solution on splanchnic oxygenation in patients undergoing cytoreductive surgery for ovarian cancer. AB - OBJECTIVE: To investigate the effectiveness of using a medium molecular weight hydroxyethyl starch solution (HES) administered as a replacement for estimated blood loss (EBL) during cytoreductive surgery for ovarian cancer on splanchnic oxygenation. METHODS: Forty-two patients undergoing cytoreductive surgery for ovarian cancer were enrolled in this prospective randomized study. As soon as the EBL exceeded 10% but was less than 20% of the estimated blood volume, the patients were randomly assigned to receive either a volume of lactated Ringer's solution (LRS) equal to three times the EBL (LRS group, n = 22) or a volume of 6% HES equal to the EBL (HES group, n = 20). Tissue oxygenation was assessed indirectly by measuring tonometric parameters, including the difference between gastric intramucosal PsCO(2) and arterial PaCO(2) (Ps-a CO(2) gap), gastric intramucosal pH (pHi) and arterial lactate acid concentration at 30 min after induction of anesthesia (baseline value), 1 hour and 2 hours after skin incision, and at the end of surgery. RESULTS: At the end of surgery,the Ps-a CO(2) gap in the HES group (8.7 +/- 1.6 mmHg) was significantly lower than that of the LRS group (18.74 +/- 4.4 mmHg, P < 0.01), while the pHi (7.30 +/- 0.05 mmHg) in the HES group was significantly higher than that of the LRS group (7.21 +/- 0.07 mmHg, P < 0.01). There was no significant difference between the two groups in terms of arterial lactate acid concentration. CONCLUSION: In patients undergoing major surgery with relatively large blood losses, volume resuscitation with medium molecular weight hydroxyethyl starch solution may improve splanchnic blood flow and tissue oxygenation. PMID- 12890371 TI - Treatment of lower urethral calculi with extracorporeal shock-wave lithotripsy and pneumatic ureteroscopic lithotripsy: a comparison of effectiveness and complications. AB - OBJECTIVE: To determine the efficacy and incidence of complications of extracorporeal shock-wave lithotripsy (ESWL) compared with pneumatic ureteroscopic lithotripsy (URSL) in the treatment of lower uretheral calculi. METHODS: From August 1997 to June 1999, 210 patients with lower ureteric calculi were treated with ESWL and the other 180 with URSL. The stones were fragmented with pneumatic lithotripter. The outcome was assessed by evacuation, retreatment and complication rates. RESULTS: ESWL for lower ureteric calculi resulted in a stone evacuation rate of 78.1%, compared with 93.3% for URSL (P < 0.05). ESWL had a retreatment rate of 11.9% and a perforation rate of 0, while URSL caused perforation of ureters in 3.3% of patients and a refreatment of 2.2%. CONCLUSION: For the management of lower ureteric calculi, ESWL provides a non-invasive, simple and safe option, and URSL has a higher stone evacuation rate but causes ureter perforation more frequently than ESWL does. Both ESWL and URSL have their respective advantages. It is recommended, however, that URSL be extensively developed for better treatment efficacy, given that the operator has an adequate technical background. PMID- 12890372 TI - Evaluation of the combination of multiple subpial transection and other techniques for treatment of intractable epilepsy. AB - OBJECTIVE: Multiple subpial transection (MST) is one approach to the surgical treatment of intractable epilepsy with epileptogenic lesion located in functional areas. To verify the effect of MST, an experimental study was performed first, followed by clinical application. METHODS: On the basis of the experimental study, MST was performed in 200 intractable epileptic patients from 1991 to 2000. Of them, 80 cases underwent MST only while 120 others underwent MST combined with other techniques, such as corpus callosotomy, temporal lobectomy and focus resection. A series of modifications of the surgical techniques were made. RESULTS: The results of the experimental study indicated that MST could inhibit the formation and spreading of epileptic discharge and limit the damage to neurons in a minimal area on the epileptogenic agent injected cortex. MST does not impair major functions of the cortex. After the clinical application and modifications, 160 patients were followed up for 1 to 8 years. Complete control of seizure was obtained in 100 cases (62.5%), significant reduction (more than 75%) in 32, reduction (more than 50%) in 20 and no change in 8. The total rate of effectiveness was 95.0%, and the significant rate of effectiveness was 82.5%. No functional defects were found in any patients. CONCLUSIONS: The results indicate that MST is an effective approach to the surgical treatment of intractable epilepsy. MST can be combined with other approaches. The outcome of the subdivision of the MST only group indicates that MST on local epileptogenic lesion without structural changes is as effective as that of the combined operation group. To evade hemispheric disturbance, MST should be done first to avoid severe complications. Hemispherectomy should be performed only on poor effected cases of MST. PMID- 12890373 TI - Transaxillary minithoracotomy in intrathoracic surgery for 316 infants and children. AB - OBJECTIVE: To introduce the technique of intrathoracic surgery performed through vertical transaxillary minithoracotomy. METHODS: From March 1989 to March 2001, 316 patients underwent intrathoracic surgery through a vertical transaxillary minithoracotomy. 285 patients suffered from patent ductus arteriosus (PDA), 10 congenital esophageal atresia, 8 congenital pulmonary cysts, 6 congenital emphysema, 1 pulmonary sequestration, 5 mediastinal tumor, and 1 eventration of the diaphragm. RESULTS: All of the patients were successfully treated under satisfactory exposure. No operative mortality and severe postoperative complications occurred. CONCLUSIONS: Intrathoracic surgery performed through a vertical transaxillary minithoracotomy appears to be less invasive, and is a simple, safe, cosmetically acceptable and efficient approach. PMID- 12890375 TI - Procedure and clinical assessments of malariotherapy: recent experience in 20 HIV patients. AB - OBJECTIVE: To demonstrate the side effects of malariotherapy and to explore safe procedures in conduct of malariotherapy for human immunodeficiency virus (HIV) infected patients. METHODS: Twenty HIV/acquired immunodeficiency syndrome (AIDS) patients were selected for the study of malariotherapy and were intravenously infected with Plasmodia vivax to induce therapeutic malaria. Malaria was terminated with chloroquine after 10 - 20 malarial febrile episodes. Clinical assessments were made before (baseline), during (malarial phase) and after (post) termination of malaria. The density of Plasmodia in peripheral blood from the HIV/AIDS patients were compared to that from HIV-negative naturally infected malarial patients who donated the blood for the therapeutically induced malaria. CD(4) cell baseline levels were correlated to the severity of malarial symptoms and parasitemia. RESULTS: There were no significant differences of Plasmodium density between the HIV/AIDS patients injected with P. vivax and the HIV-negative blood donors. However, it was found that the HIV-positive patients had milder malarial symptoms and parasitemia with progressively lower CD(4) cell baseline levels. All patients developed every day or every other day fever episodes with headache and shaking chill. These symptoms were well tolerated with the aid of anti-pyretic medications. Spleen and liver enlargement were seen in 15 of 20 and 4 of 20 patients respectively. Transitory liver effects with increase of serum glutamic-pyruvic transaminase were seen in 2 of 20 during malarial phase. Most patients experienced mild to medium anemia and 6 of 20 patients developed thrombocytopenia during malarial phase. All these side effects disappeared after termination of malaria or within one month thereafter. No complications occurred in these patients. CONCLUSIONS: Therapeutically induced acute vivax malaria was well tolerated in 20 HIV-positive subjects who represented a range of CD(4) cell levels from 1868/ micro l to 15/ micro l. Malariotherapy did not induce complications while increasing CD(4) cell levels in most treated HIV/AIDS patients (results published elsewhere). PMID- 12890374 TI - Inhaling beta(2)-agonist with heliox-driven in bronchial asthma. AB - OBJECTIVE: To evaluate the effectiveness of a helium-oxygen mixture (79%He- 21%O(2)) as an aerosolizing compressed gas for beta(2)-agonist therapy in patients with an asthma exacerbation. METHODS: Twenty-four patients in the outpatient department with a mild to moderate exacerbation of asthma were enrolled. The patients were randomly divided into an experimental group (13 cases) and a control group (11 cases). The experimental group inhaled Berotec with heliox-driven, and the control group inhaled Berotec with compressed air driven. Eight hospitalized patients in the respiratory department with severe exacerbation of asthma were enrolled. The patients inhaled Berotec with heliox driven or compressed air-driven in a random order. RESULTS: The results of spirometric parameters and arterial blood-gas analysis were measured. In the mild to moderate asthma patients, no statistical differences between the two groups for forced vital capacity (FVC), forced expired volume in one second (FEV(1)), and expiratory flow in 50% forced vital capacity (FEF(50)) were presented. But the severe patients showed significant differences between heliox-driven and compressed air-driven for FVC, FEV(1), FEF(50) and partial pressure of oxygen (PaO(2)). CONCLUSIONS: Compared with the traditional inhalation of beta(2) agonist therapy using compressed air-driven, the method of inhaling beta(2) agonist with heliox-driven has more obvious benefits for those suffering from severe asthma. This is likely due to the cooperative effects between inhaling heliox on its physical gas properties and improving delivery of beta(2)-agonist in the treatment of exacerbation of severe asthma. PMID- 12890376 TI - Epstein-Barr virus encoded latent membrane protein 1 induces TRAF1 expression to promote anti-apoptosis activity via NF-kappaB signaling pathway in nasopharyngeal carcinoma. AB - OBJECTIVES: To identify whether Epstein-Barr virus (EBV) encoded latent membrane protein 1 (LMP1) can induce tumor necrosis factor receptor-associated factor 1 (TRAF1) expression and promote its anti-apoptosis activity via the NF-kappaB signaling pathway, and assess that LMP1 suppresses apoptosis in nasopharyngeal carcinoma (NPC). METHODS: A stable transfected cell line HNE2-LMP1 was established by introducing LMP1 cDNA into HNE2 cells. Transactivation of TRAF1 was determined by luciferase reporter assay, while expression of TRAF1 mRNA was detected by RT-PCR and expression of TRAF1 protein and caspase 3 by Western blot analysis. Apoptosis activity was observed through fluorescence staining. RESULTS: LMP1 induced TRAF1 expression in NPC cells and caused a decrease in apoptosis. This induction could be blocked by antisense LMP1. Moreover, LMP1-mediated induction of a TRAF1 promoter-driven reporter gene was significantly impaired when the kappaB site kappaB1 or kappaB5 was disrupted, whereas mutation of kappaB3 had only a minor effect on LMP1 dependent up-regulation of the reporter gene. CONCLUSION: LMP1 induces TRAF1 expression and promotes its anti-apoptosis activity via the NF-kappaB signaling pathway, which may be one of the mechanisms that LMP1 uses to suppress apoptosis in NPC cells. PMID- 12890377 TI - Effect of human hepatocyte growth factor on promoting wound healing and preventing scar formation by adenovirus-mediated gene transfer. AB - OBJECTIVE: To evaluate the effects of hepatocyte growth factor (HGF) on the prevention of scar formation and the promotion of wound healing by gene transfer. METHODS: A total of 12 female New Zealand rabbits were used in this study. Rabbits were anesthetized with an intravenous injection of sodium pentobarbital, and identical wounds were made over the ventral surface of each ear. Five circular wounds, 7 mm in diameter, were created in each ear by excision through the skin to the underlying cartilage using sterile technique. After the surgical procedures, 10 of the rabbits were randomly allocated to five groups, with 2 rabbits in each group: Ad-HGF group 1, Ad-HGF group 2, Ad-HGF group 3, Ad-GFP (a reporter gene) group and the solvent group. Immediately after surgery, 6 x 10(7) pfu Ad-HGF, 6 x 10(8) pfu Ad-HGF, 6 x 10(9) pfu of Ad-HGF, 6 x 10(9) pfu of Ad GFP, or same volume of solvent (PBS, pH 7.2) was applied once to each wound in groups 1 to 5, respectively. One additional rabbit was used to evaluate the transfer efficiency of the adenovirus vector by transferring Ad-GFP (6 x 10(9) pfu) into its wounds. Ice slides of wounds from this animal were observed under fluorescence microscopy. Another additional rabbit was used to evaluate the expression of HGF and TGFbeta1 after transferring Ad-HGF (6 x 10(9) pfu) into each of its wound. Immunohistochemistry was used for detection. RESULTS: The effect of HGF on reducing excessive dermal scarring was observed by adenovirus mediated gene transfer. Transfection of the human HGF cDNA into skin wounds through an adenoviral vector suppressed the over-expression of TGFbeta1, which plays an essential role in the progression of dermal fibrogenesis. Application of HGF to the wounds significantly enhanced wound healing and inhibited over scarring. CONCLUSION: HGF gene therapy could be a new approach for preventing excessive dermal scarring in wound healing. PMID- 12890378 TI - Caspase-3 and its inhibitor Ac-DEVD-CHO in rat lens epithelial cell apoptosis induced by hydrogen in vitro. AB - OBJECTIVE: To investigate the role of caspase-3 and its inhibitor Ac-DEVD-CHO in rat lens epithelial cell apoptosis induced by hydrogen peroxide (H(2)O(2)) in vitro. METHODS: Rat lenses were incubated in modified Eagle's medium containing 2 mmol/L H(2)O(2) to induce apoptosis in vitro. Apoptosis in lens epithelial cells was assessed by transmission electron microscopy and annexin V-propidium iodide (PI) double staining flow cytometry after 12, 24 and 48 h of incubation. The activity of caspase-3 was analyzed by western blotting. RESULTS: Observations under transmission electron microscopy revealed that 2 mmol/L H(2)O(2) could effectively induce lens epithelial cell apoptosis in vitro. Caspase-3 activity increased during cell apoptosis and the peak measurement occurred at 24 h after treatment with H(2)O(2). Cell apoptosis was blocked by caspase-3 inhibitor Ac DEVD-CHO. CONCLUSIONS: The activation of caspase-3 plays an important role in executing apoptosis in H(2)O(2)-treated lens epithelial cells and in the formation of cataract. The caspase-3 inhibitor Ac-DEVD-CHO may effectively prevent lens epithelial cell apoptosis caused by oxidative injury. PMID- 12890379 TI - Localization of the human language cortex by magnetic source imaging. AB - OBJECTIVE: To localize the language cortex associated with Chinese word processing by magnetic source imaging (MSI). METHODS: Eight right-handed and one left-handed healthy native Chinese subjects were examined by magnetoencephalography (MEG) and a 1.5T magnetic resonance imaging (MRI) unit. All subjects were given pure tone stimuli 50 times, 150 pairs of Chinese words (meaning related or unrelated) auditory stimuli, and pure tone stimuli subsequently 50 times. Evoked response fields time locked to the pure tone and Chinese words were recorded using a whole-head neuromagnetometer in real-time. The acquired data were averaged by the acquisition computer according to the response to the pure tone, related pairs of words and unrelated pairs of words. The data obtained by MEG were superimposed on MRI, using a GE Signa 1.5T system. RESULTS: MEG, showed there were two obviously higher magnetic waves named M50 and M100, which were localized in the bilateral transverse temporal gyri in all subjects. The responses to the pairs of Chinese words (meaning related or unrelated) were similar in the same hemisphere of the same subjects. There was a higher peak during 300 - 600 ms in the right hemisphere of one left handed subject, but no peak in the left hemisphere, indicating that the language dominant hemisphere was localized in the right hemisphere. Superimposing the MEG data on MRI, the language area was localized in the Wernicke's areas. A 300 - 600 ms response peak was obsarved in each hemisphere (the amplitude of the 300 - 600 ms response peak in each hemisphere was almost the same) in two right-handed subjects, showing that the language area was localized in the 2 hemispheres in the two subjects. There was one peak in each hemisphere (300 - 600 ms response) in 6 subjects, but the amplitude of the wave in the left hemisphere in the 6 subjects was much higher than that in the right hemisphere. By choosing randomly from the later component (300 - 600 ms response) several time points and superimposing them on MRI, all time points were localized in the posterior part of the superior temporal gyri, which is the Wernicke's areas. CONCLUSIONS: Comparing with the later component of the bilateral hemispheres, the wave amplitude in the language dominant hemisphere was much higher than that in the non-language dominant hemisphere. The language areas could be identified by judging whether meanings of pairs of Chinese words were related or not. PMID- 12890381 TI - Diagnosis and surgical treatment of ruptured aneurysm in sinus of Valsalva. AB - OBJECTIVE: To evaluate the methods used to diagnose and surgically treat ruptured aneurysm in sinus of Valsalva (RASV). METHODS: Thirty-seven hospitalized patients with ruptured aneurysms in the sinus of Valsalva from September 1981 to April 2001, including 21 cases (56.7%) of RASV associated with ventricular septal defects (VSD) and 11 (29.7%) with aortic valvular prolapse were given surgical interventions. Under hypothermia and extracorporeal circulation, we successfully performed the surgical correction of RASV for all 37 patients VSD repair in 21 patients, aortic valvuloplasty in 6 and aortic valvular replacement in 2. RESULTS: There was no hospital deaths among these patients, although residual shunting occurred in two patients and acute renal failure was found one. Follow up study of one month to 20 years in the patients undergoing repair of RASV revealed that the mostly individuals treated with operation obtained satisfactory cardiac function. CONCLUSION: Correct diagnosis of ruptured aneurysm in sinus of Valsalva should be confirmed immediately and surgical correction should be carried out as soon as possible. PMID- 12890380 TI - FBN1 mutation in Chinese patients with Marfan syndrome and its gene diagnosis using haplotype linkage analysis. AB - OBJECTIVES: To analyze the FBN1 mutations in Chinese patients with Marfan syndrome (MFS) and to make a genetic diagnosis based on haplotype linkage analysis for MFS. METHODS: Nine MFS families (17 patients) were analyzed with single strand conformation polymorphism (SSCP) and sequencing. Four primers were designed for the flanking sequences of FBN1 gene and used for haplotype segregation analysis of MFS(B). RESULTS: SSCP band alteration was detected in the PCR products for exon 25 in MFS(A) II:1. Direct sequencing revealed a small 13 bp deletion; the deleted sequence is gccTc Tgcaccca at bases 3243-3456 of the cDNA in exon 25. This mutation was novel. MFS(B) families were analyzed using the haplotype linkage technique. The data suggested that MFS(B) families were linked to the FBN1 gene. The proband's daughter was an asymptomatic patient. CONCLUSION: The combination of mutation detection and chromosome haplotype analysis can provide better evidence for a genetic diagnosis of MFS. PMID- 12890382 TI - Single photon emission CT perfusion imaging of cerebral blood flow of early syphilis patients. AB - OBJECTIVE: To injvestigate the cerebral blood flow of patients with early syphilis. METHODS: (99)Tc(m)-ECD as brain perfusion imaging agent was used in single photon emission computed tomography (SPECT) for 32 patients with early syphilis and 15 controls. Visual analyses were made on every BSPECT image. RESULTS: The 32 patients with early syphilis had general, patchy hypoperfusion of cerebral blood flow. Fourteen of the 32 patients had 48 episodes of marked patchy hypoperfusion of rCBF. The responsible areas of hypoperfusion in a patchy distribution involved the left frontal lobe (6 episodes), right frontal lobe (3), left parietal lobe (7), right parietal lobe (6), left temporal lobe (11), right temporal lobe (5), left occipital lobe (3), left basal ganglia (3), cerebellum (1), and nerve nuceus (1). No abnormality was found in the control group. CONCLUSIONS: Cerebral blood flow abnormalities exist in patients with early syphilis. General patchy hypoperfusion on SPECT imaging is common. PMID- 12890383 TI - Involvement of dynorphin A in the inhibition of morphine physical dependence by N nitro-L-arginine in rats. AB - OBJECTIVE: To investigate the involvement of immunoreactive-dynorphin A in the inhibitory effect of N-nitro-L-arginine on the morphine physical dependence in rats. METHODS: The rats were rendered dependent on morphine by subcutaneous administration of morphine solution three times daily in a manner of dose increment of 5 mg.kg(-1) for 6 days. The degree of morphine physical dependence was monitored by scoring the abstinence syndromes precipitated by 5 mg.kg(-1) naloxone of the rats. The expression levels of immunoreactive dynorphin A in tissues were determined using a radioimmunoassay. RESULTS: Intraperitoneal injection of 5 mg.kg(-1) N-nitro-L-arginine suppresses most of the withdrawal symptoms of morphine dependent rats. N-nitro-L-arginine can elevate the expression of immunoreactive dynorphin. CONCLUSIONS: Chronic N-nitro-L-arginine administration can inhibit the development of morphine physical dependence in a manner of dose-dependence, which is significantly related to its role of regulating the endogeneous dynorphin system. PMID- 12890384 TI - Cardioprotective effects of morphine on rat heart suffering from ischemia and reperfusion. AB - OBJECTIVE: To investigate the cardioprotective effects of morphine on ischemic reperfused rat heart in vitro and its mechanism. METHODS: The isolated rat heart was perfused in a Langendorff apparatus. Infarct myocardium was determined by TTC. Coronary flow (CF), heart rate (HR), left ventricular pressure (LVP), the first derivative of ventricular pressure (LVP/dtmax) and infarct size after ischemia and reperfusion in rat heart given 0.3 micro mol/L morphine were observed. The effects of naloxone and glibenclamide on the cardioprotection of morphine were also measured. RESULTS: After ischemia and reperfusion, CF, HR, LVP and LVP/dtmax of isolated rat hearts decreased significantly (P < 0.01). After morphine preconditioning, HR, LVP and LVP/dtmax increased (P < 0.01) and infarct size was reduced significantly (P < 0.01), while no significant change in CF (P > 0.05). The cardioprotective effects of morphine were abolished by naloxone or glibenclamide completely. CONCLUSIONS: Morphine can reduce ischemia-reperfusion injuries in isolated rat heart. The cardioprotective effects of morphine are mediated by a local opioid receptor-K(ATP) channel linked mechanism in rat hearts. PMID- 12890385 TI - A correlative study between AQP4 expression and the manifestation of DWI after the acute ischemic brain edema in rats. AB - OBJECTIVE: To investigate the rule of the aquaporin-4 (AQP4) expression in acute ischemic brain edema, and to study the correlation between AQP4 expression and diffusion-weighted imaging (DWI). METHODS: Thirty-six Wistar rats were divided into 2 groups randomly, control group (n = 12) and operation group (n = 24) in which right middle cerebral artery of each animal had been occluded unilaterally (MCAO) at interval times of: 15 minutes, 30 minutes, 1 hours, 3 hours, 6 hours and 24 hours, respectively. The operation process of the control group was the same as the operation group except for the MCAO. All groups were examined using DWI. The apparent diffusion coefficient (ADC), relative density (rd) and relative area (rs) of the biggest hyperintensity signal layer on DWI were measured. After that the animals were sacrificed and perfused with the mixture solution consisting of TTC. The biggest layers of the ischemic cerebral tissues in each rat corresponding to the DWI were stained with TTC and examined with immunochemistry (DeltaS), in situ hybridization (alpha) and histology. RESULTS: There was no significant change in the control group. In the operation group, a hyperintensity signal was found in the DWI of the right MAC territory at 15 minutes after MCAO. The ADC value decreased quickly within one hour after MCAO, while the AQP4 expression, rd-DWI and rs-DWI increased rapidly during this stage. As time progressed, the ADC value decreased further to (2.1 +/- 0.6) x 10(-4) mm(2)/s at 3 hours, and then began to increase slowly till 24 hours. But the AQP4 expression (DeltaS and alpha) and rd as well as the rs continuously increased slowly between 1 hour and 6 hours after MCAO, followed a peak after 6 hours. The AQP4 expression (alpha) showed a positive relationship with the rs-DWI, they all presented two peaks and a plateau. The corresponding sequential pathologic changes were a gradual increase of intracellular edema (within one hour), then an emergence of vasogenic edema (1-6 hours), and final necrosis and liquefaction (6 24 hours). CONCLUSIONS: Upregulated expression of AQP4 may play a significant role in acute ischemic brain edema, especially during the stages of intracellular edema and necrosis, but it has no correlation to vasogenic edema. Certainly, the high expression of AQP4 is perhaps one of the most important reasons of the decrease of ADC and hyperintensity on the DWI in the intracellular edema. PMID- 12890386 TI - Using a cDNA microarray to study cellular gene expression altered by Mycobacterium tuberculosis. AB - OBJECTIVE: To examine the global effects of Mycobacterium tuberculosis (M. tuberculosis) infection on macrophages. METHODS: The gene expression profiling of macrophage U937, in response to infection with M. tuberculosis H(37)R(a), was monitored using a high-density cDNA microarray. RESULTS: M. tuberculosis infection caused 463 differentially expressed genes, of which 366 genes are known genes registered in the Gene Bank. These genes function in various cellular processes including intracellular signalling, cytoskeletal rearrangement, apoptosis, transcriptional regulation, cell surface receptors, cell-mediated immunity as well as a variety of cellular metabolic pathways, and may play key roles in M. tuberculosis infection and intracellular survival. CONCLUSIONS: M. tuberculosis infection alters the expression of host-cell genes, and these genes will provide a foundation for understanding the infection process of M. tuberculosis. The cDNA microarray is a powerful tool for studying pathogen-host cell interaction. PMID- 12890387 TI - Gene expression profile changes in NB4 cells induced by realgar. AB - OBJECTIVES: To compare the gene expression profiles of acute promyelocytic leukemia cell line NB(4) before and after 12 hours of realgar treatment using cDNA microarray. METHODS: Two cDNA probes were prepared through reverse transcription from mRNA of both untreated and realgar treated NB(4) cells. The probes were labeled with Cy3 and Cy5 fluorescence dyes individually, hybridized with cDNA microarray representing 1003 different human genes, and scanned for fluorescent intensity. The genes were screened through the analysis of the difference in two gene expression profiles. RESULTS: The analysis of gene expression profiles indicates that 9 genes were up-regulated and 37 genes were down-regulated. Among the 9 up-regulated genes, 2 genes were involved in a proteasome degradation pathway. Some genes related to protein synthesis, signal transduction and cell receptors were down-regulated. CONCLUSION: PSMC2 and PSMD1 genes may play an important role in the apoptosis and partial differentiation of NB(4) cells. PMID- 12890388 TI - Association between HLA-DQA1, -DQB1 gene polymorphisms and susceptibility to asthma in northern Chinese subjects. AB - OBJECTIVE: Human leukocyte antigen (HLA) class II genes, especially HLA-DQ genes, which are highly polymorphic, have been thought to be candidate loci for the etiology of asthma, and shown to be involved in antigenic presentation. This study was conducted to investigate whether susceptibility or resistance to asthma is associated with HLA-DQA1 and DQB1 genes polymorphism. METHODS: Venous blood samples were collected from northern Chinese population with Han ethnic. (1) One hundred and twenty-five unrelated asthmatic individuals and 52 subjects from 12 asthmatic pedigrees. (2) Ninety-six healthy controls without asthma and atopy with the same ethnic. Genomic DNA was extracted using standard phenol-chloroform method. The second exon of HLA-DQA1 and DQB1 genes were amplified by sequence specific primer-polymerase chain reaction (SSP-PCR) method. All asthmatics had their serum IgE (total and specific) antibody or skin-prick test measured, bronchial reactivity to methacholine (Mch) and bronchial reversibility by beta(2) agonist evaluated. RESULTS: HLA-DQA1 * 0104 allele and HLA-DQB1 * 0201 allele were significantly higher in asthmatics than those in healthy controls (0.408 vs 0.177, P < 0.01; 0.568 vs 0.198, P < 0.01). Odds ratios (ORs) were 3.203 (95% CI 1.699 - 6.037), 5.328 (95% CI 2.883 - 9.849) respectively. Conversely, HLA-DQA1 * 0301 allele and HLA-DQB1 * 0301 were significantly decreased in asthmatics compared to healthy controls (0.296 vs 0.50, P < 0.01; 0.4 vs 0.563, P < 0.05); Logistic regression analysis showed that HLA-DQA1 * 0104 allele was associated independently with asthma etiology, OR [represented by Exp(B)] was 5.0942 with 95% CI 2.2520 - 21.1813; Spearman's analysis showed that HLA-DQA1 * 0104 allele and HLA-DQB1 * 0201 allele were positively associated with atopy, the correlation coefficient were 0.183 and 0.289 respectively, P < 0.01. By contrast, HLA-DQA1 * 0301 allele was negatively related to atopy, the correlation coefficient was 0.168, P < 0.05; linkage analysis did not support the view that HLA-DQA1/DQB1 genes were linked to asthma with LOD value being 0.72. CONCLUSIONS: HLA-DQA1 * 0104 allele and HLA-DQB1 * 0201 allele were implicated in susceptibility to asthma and atopy, HLA-DQA1 * 0301 allele and HLA-DQB1 * 0301 might be protective factor against asthma. Asthma and atopy are multifactorial disorders, HLA-DQA1 and DQB1 genes are involved in the regulation of immune specific response to common allergen. PMID- 12890389 TI - Relationship between alterations of p16(INK4a) and p14(ARF) genes of CDKN2A locus and gastric carcinogenesis. AB - OBJECTIVE: To investigate the relationship between alterations of p16(INK4a) and p14(ARF) genes and gastric carcinogenesis. METHODS: The tumors and neighboring gastric tissues from 48 patients with gastric cancer were studied. The homozygous deletion, mutation, methylation of the CpG islands, and mRNA expression of p16(INK4a) and p14(ARF) genes were assessed by PCR, PCR-SSCP, PCR based methylation assay, and RT-PCR. RESULTS: (1) The homozygous deletion rate of p16(INK4a) and p14(ARF) was 35.4% (17/48), and no homozygous deletion was examined in any gastric tissue neighboring the tumor. (2) There was no point mutation of p16(INK4a) and p14(ARF) in 31 gastric cancers without homozygous deletion or in the matched gastric tissues adjacent to the tumor. (3) Methylation of the CpG islands of p16(INK4a) and p14(ARF) was detected in 47.9% (23/48) of gastric cancers, while methylation was observed only in 2 of 48 gastric tissues neighboring the cancer with a significant difference (P < 0.01). (4) The loss rate of p16(INK4a) mRNA was 47.9% (23/48) in gastric cancer, and the patients of the combined methylation of exons 1alpha and 2 had a higher loss rate (100%, 6/6) of p16(INK4a) mRNA than those of the methylation of the other exons (11.8%, 2/17, P < 0.01); the loss rate of p14(ARF) mRNA was 45.8% (22/48) in gastric cancer, and patients with the combined methylation of exons 1beta and 2 had a higher loss rate (100%, 3/3) of p14(ARF) mRNA than those of the methylation of the other exons (15%, 3/20, P < 0.05). (5) The combined loss of p16(INK4a) and p14(ARF) mRNAs was examined in 1 (5.6%) of 18 patients of well and moderately differentiated carcinomas, and 11 (36.7%) of 30 patients of poorly and not differentiated carcinomas with a significant difference (P < 0.05). CONCLUSION: p16(INK4a) and p14(ARF) genes are frequently inactivated by homozygous deletion and methylation of the 5'CpG islands in gastric cancer, which may play an important role in the carcinogenesis of gastric cancer. PMID- 12890390 TI - Influence of skeletal muscle satellite cells implanted into infarcted myocardium on remnant myocyte volumes. AB - OBJECTIVE: To study the effects of skeletal muscle satellite cells implanted into infarcted myocardium on the volume of remnant myocytes. METHODS: Thirty-six adult mongrel canines were divided randomly into implantation group and control group. In the implantation group, skeletal muscle satellite cells taken from the gluteus maximus muscles of the dogs were cultured, proliferated and labeled with 4',6 diamidino-2-phenylindone (DAPI) in vitro. In both groups, a model of acute myocardial infarction was established in every dog. In the implantation group, each dog was injected with M199 solution containing autologous skeletal muscle satellite cells. The dogs in the control group received M199 solution without skeletal muscle satellite cells. The dogs of both groups were killed 2, 4 and 8 weeks after implantation (six dogs in a separate group each time). Both infarcted myocardium and normal myocytes distal from the infracted regions isolated were observed under optical and fluorescent microscope. Their volumes were determined using a confocal microscopy image analysis system and analyzed using SAS. A P < 0.05 was considered significant. RESULTS: A portion of the implanted cells differentiated into muscle fiber with striations and were connected with intercalated discs. Cross-sectional area and cell volume were increased in normal myocardium. Hypertrophy of remnant myocytes in the infarcted site after skeletal muscle cell implantation was much more evident than in the control group. Cross sectional area, cell area and cell volume differed significantly from those of the control group (P < 0.05). Hypertrophy of the cells occurred predominantly in terms of width and thickness, whereas cell length remained unchanged. CONCLUSION: Skeletal muscle satellite cells implanted into infarct myocardium, could induce the hypertrophy of remnant myocyte cells in the infarcted site and could also aid in the recovery of the contractile force of the infarcted myocardium. PMID- 12890391 TI - Human DNA contains sequences homologous to the 5'-non-coding region of hepatits C virus: characterization with restriction endonucleases reveals individual varieties. AB - OBJECTIVE: To investigate a 272 base pair section of the 5'-non-coding region of genomic DNA from the peripheral blood monounuclear cells of healthy hepatitis virus C (HCV)-negative human subjects (not patients). METHODS: This sequence section bears interest because (1) it harbors several potential methylation (Cp rich) sites, and (2) it represents the largest part of its internal ribosomal entry site. A pre-PCR digestion protocol was established making consistent use of four restriction endonucleases selected for certain features: SmaI, XmaCI, MspI, and HpaII are inhibited if methylation(s) are present at certain cytosines within their cutting sequences. RESULTS: The suspected HCV-specific sequence was found in the DNA of each subject tested. The pre-PCR digestion assay reveals individual differences in their pattern of methylation, which may be due to possible epigenetic phenomena. CONCLUSIONS: The results provide formal proof that these HCV-specific sequences are contained in the genomic or extra chromosomal target DNA, and probably belong to a new class of endogenous sequences. PMID- 12890392 TI - Effects of ursodeoxycholic acid on intrahepatic cholestasis in rats. AB - OBJECTIVE: To investigate the effects of treatment with ursodeoxycholic acid (UDCA) on intrahepatic cholestasis in rats, and to explore its mechanism. METHOD: Rats suffering from intrahepatic cholestasis were treated with UDCA. Their serum alanine transaminase (ALT), alkaline phosphatase (ALP), total bilirubin (TBIL), direct bilirubin (DBIL), gamma-glutamyl transpeptidase (gamma-GT), total cholesterol (TCH), bile flow, total bile acid excretion, total Na(+) and TCH of bile were measured before and after treatment. In addition, the changes of liver tissue under microscrope were observed and recorded. RESULTS: Compared with the control group, serum ALT, ALP, TBIL, DBIL, gamma-GT and TCH of rats in the treatment group decreased, while bile flow, total bile acid excretion, total Na(+) and TCH decreased significantly. Degeneration of hepatocytes, infiltration of inflamed cells and proliferation of small bile ducts in the treatment group were improved under microscope. CONCLUSION: UDCA may have therapeutic effects on cholestatic hepatitis. The mechanism may involve in its hydrophilicity, choleretic effect and immune modulation. PMID- 12890393 TI - Clinical and imaging findings in patients with severe acute respiratory syndrome. AB - OBJECTIVE: To investigate the clinical and imaging characteristics of severe acute respiratory syndrome (SARS), and to study their relationship. METHODS: Forty-six SARS confirmed patients were admitted to our hospital from February to April, 2003. X-ray examination documents were available in all cases and chest CT scanning was acquired in 6 cases, which were analyzed retrospectively, accompanied by their clinical features. RESULTS: Fever was found in 97.8% of the patients. Clinical symptoms were mild, but X-ray and CT findings were distinct. CT scanning demonstrated ground glass like lesions and large patchy exudation and consolidation at the early stage in 6 cases. Different findings on radiography and CT were related to the different phases of the disease. After treatment, most lesions were absorbed completely, but slowly in patients with multi-lobe consolidation and/or extensive interstitial infiltration. CONCLUSION: Special clinical and imaging findings could be found in SARS cases. The prognosis of SARS patients is related to the degree of lesions detected by radiography and CT. PMID- 12890394 TI - Advantages of using an image-guided system for transnasal endoscopic surgery. AB - OBJECTIVE: To evaluate the advantages of image-guided system in transnasal endoscopic surgery. METHODS: Transnasal endoscopic surgery was performed with the aid of an image-guided system in 28 patients, supported with histopathologic diagnoses of chronic sinusitis with/without nasal polyps (10 cases), juvenile nasopharyngeal angiofibroma (4 cases), pituitary adenoma (6 cases), ethmoidal ossifying fibroma (3 cases), nasopharyngeal mixed tumor (2 cases), nasal leiomyoma (1 case), fungal sinusitis (1 case) and inverted nasal papilloma (1 case). RESULTS: For all the patients, the time periods from initialization to surgery ranged from 15 to 30 minutes (a mean of 26 minutes). The calibration coefficient ranged from 1.3 to 2.0. Accuracy of localization fell within 1 mm. Compared with traditional endoscopic surgery, operation times were not noticeably different. No complications occurred. CONCLUSIONS: The image-guided system was able to identify borders and critical anatomical structures in real-time, especially of those with distorted anatomical markers. It provided a powerful means for a safer and less invasive endoscopic sinus surgery. PMID- 12890395 TI - The relationship between catecholamines levels in mother and fetus, and pathogenesis of pregnancy-induced hypertension. AB - OBJECTIVE: To study the relationship between pregnancy-induced hypertension (PIH) and catecholamine levels. METHODS: Catecholamines levels in maternal and fetal blood were determined in 116 patients with PIH and 40 normal control subjects using high performance liquid chromatography. The normal control subjects and PIH cases were selected from patients at term pregnancy receiving elective cesarean section. RESULTS: Plasma norepinephrine (NE) levels were significantly higher in patients with severe PIH than those in control subjects (P < 0.05). Both patients and control subjects had higher NE levels in the umbilical artery blood than in the umbilical vein blood (P < 0.05). NE levels in the umbilical artery blood were five times higher than those in the maternal blood. CONCLUSION: The pathogenesis of PIH may relate to catecholamine concentrations in fetus. PMID- 12890396 TI - Ethnic-specific splicing mutation of the carnitine-acylcarnitine translocase gene in a Chinese neonate presenting with sudden unexpected death. PMID- 12890397 TI - A tiny dural arteriovenous fistula. PMID- 12890398 TI - Highly efficient expression, purification of recombinant LTB protein and its activity against mucosal immunoadjuvant by nasal immunization. AB - OBJECTIVE: To develop an efficient expression, purification system of recombinant Escherichia coli heat-labile enterotoxin B subunit (rLTB) and study its activity against mucosal immunoadjuvant by nasal immunization. METHODS: A recombinant, pMMB68-LTB was generated by cloning the LTB cDNA fragment into an expression vector (pMMB68) and transformed it into the host strain marine vibrio VSP60. The relevant target protein was identified using SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot. Sephacryl S-100 gel filtration chromatography was carried out for purification of rLTB in engineering bacteria VSP60. BALB/c mice received hen egg lysozyme (HEL) alone or combined with rLTB by nasal administration. After three times immunization, IgG and IgA antibody levels in serum or small intestine wash samples were determined using ELISA. RESULTS: rLTB protein was highly expressed in VSP60. After gel filtration with Sephacryl S 100, the purity of rLTB reached 98.1%, the yield rate was about 52%. After immunization, IgG and IgA antibody responses specific to HEL in system and mucosa of HEL + rLTB groups were significantly increased, compared with the HEL alone group (P < 0.001). CONCLUSIONS: A set of protocols for large-scale rLTB preparation has been established, which is simple, efficient and applicable. The rLTB protein we prepared was proved to be a powerful mucocal adjuvant, which could greatly enhance systemic and mucosal immune responses to nasally co administered antigen. PMID- 12890399 TI - [Early diagnosis of airway inflammation in smokers: still a challenge]. PMID- 12890400 TI - [Community-acquired pneumonia requiring hospitalization in immunocompetent elderly patients: clinical features, prognostic factors and treatment]. AB - BACKGROUND: Community-acquired pneumonia (CAP) is the leading cause of death from infectious disease among the elderly. This study was carried out to examine the clinical manifestations, etiology, prognostic factors and treatment of CAP in immunocompetent elderly patients requiring hospitalization. RESULTS: A total of 306 elderly (80 +/- 7 years) patients were evaluated: 54% were male, 89% had concurrent diseases (principally cardiovascular and neurological disease, chronic obstructive lung disease and diabetes), and 97% were treated with second-or third generation cephalosporins. Mean hospital stay was 10 days, and mortality was approximately 10% in hospital and around 13% on follow up at 30 days. As compared to younger CAP patients, multiple comorbidity, altered mental status, hypoxemia, high serum urea nitrogen on hospital admission were more frequent in the elderly. Intermediate care and intensive care unit admissions were also more frequent in the elderly. Hospital length of stay as well as mortality in the hospital and at 30 days were higher in elderly patients. The pathogen was identified as part of routine care in around 25% of cases. The most frequent pathogens were Streptococcus pneumoniae (10.5%), enteric Gram negative bacilli (5.2%), Staphylococcus aureus (4.2%) and Haemophilus influenzae (3.9%). In multivariate analysis the prognostic factors on admission associated with in-hospital mortality were advanced age (> 83 years), absence of cough, low blood pressure and hyperphosphatemia. CONCLUSION: CAP in elderly patients is a prevalent disease with specific clinical and epidemiological characteristics, clinical course and prognosis. PMID- 12890401 TI - [Multicenter, prospective study comparing enoxaparin with unfractionated heparin in the treatment of submassive pulmonary thromboembolism]. AB - OBJECTIVE: To compare the safety and efficacy of enoxaparin and unfractionated heparin in the treatment of submassive pulmonary thromboembolism (PTE). MATERIAL AND METHODS: Fifty-six patients with PTE who did not need fibrinolytic treatment were enrolled prospectively. The patients were randomly assigned to 2 treatment groups: Group A received enoxaparin (1 mg/kg every 12 hours) and Group B received adjusted doses of unfractionated heparin. The oral anticoagulant therapy was started on confirmation of the diagnosis and continued for 6 months. Incidences of recurrence of thromboembolism and of severe bleeding were assessed at the end of this period. RESULTS: Six patients were withdrawn from the study. Twenty-nine of the 50 remaining patients were in Group A (enoxaparin) and 21 in Group B (unfractionated heparin). A recurrence of thromboembolism was diagnosed in 3 patients from Group A (10.7%) and 2 patients from Group B (9.5%). There were no significant differences. Two patients died, one death being attributed to bleeding secondary to the oral anticoagulant treatment (Group A) and the other to a process unrelated to PTE. CONCLUSIONS: Enoxaparin seems to be as effective and safe as unfractionated heparin in the initial treatment of PTE. PMID- 12890402 TI - [Individually tailored medical counseling for pregnant smokers]. AB - OBJECTIVES: To identify the characteristics of smoking addiction in a group of pregnant women, to evaluate the efficacy of medical counseling tailored to the patient's stage in the cessation process and to examine factors that might affect the ability of a woman to quit smoking during pregnancy. PATIENTS AND METHODS: One hundred sixteen women (mean age 29.71 5.44 years; range 15-41) were referred by obstetricians to a respiratory medicine specialist if they continued smoking after being advised to quit. The information each woman received was appropriate to her stage in the cessation process and covered the risks that smoking posed for her child and the benefits that would come from quitting. Each woman was provided with guidelines for quitting and helped to choose the first day to start the cessation process. The women received follow-up counseling sessions after the first and third months. RESULTS: Twenty-seven women (23.3%) quit smoking, 18 (15.52%) advanced a stage in the cessation process and 30 (25.86%) smoked less. Overall, the program benefited 60 women (51.7%) who quit smoking, reduced the number of cigarettes they smoked or progressed to a new stage. CONCLUSIONS: The results reveal that most pregnant smokers need the intervention of specialists with knowledge of smoking addiction to facilitate the cessation process. PMID- 12890403 TI - [Prognostic factors of early morbidity and mortality after lung transplantation]. AB - OBJECTIVES: Despite years of experience with lung transplantation, the rate of perioperative mortality remains high. The objective of this study was to look at our experience in the early postoperative period following lung transplantation in an effort to identify possible pre-, intra- and postoperative risk factors associated with mortality. PATIENTS AND METHODS: A retrospective study of 68 consecutive patients receiving lung transplants over a period of 56 months. The conditions that led to transplantation were obstructive disease (40%), interstitial disease (33%) and suppurative disease (27%). Pre-, intra- and postoperative characteristics of donors and recipients were analyzed for their relation to morbidity and mortality. Statistical studies were done using SPSS 10.0 software. A p-value less than.05 was considered significant. Univariate analysis identified variables associated with the incidence of mortality in the postoperative recovery unit, and the variables with statistically significant associations were entered into multivariate analysis, using a logistic regression model to calculate odds ratio (OR) and 95% confidence intervals (CI). RESULTS: No donor variables correlated with mortality. Patients with suppurative lung disease had a lower mortality rate (0% vs 30%; P = 0.04). Mortality was related to ischemic time longer than 300 minutes (OR = 2) and the use of extracorporeal circulation (OR = 4). A PaO2/FiO2 ratio less than 150 during the first 24 hours following transplantation (OR = 5) and reoperation due to bleeding (OR = 12) were the variables showing the highest correlations with mortality during the early postoperative period. CONCLUSIONS: The mortality rate during the early postoperative period in our series was 22%. The survival rate was better in patients with suppurative lung disease. Bleeding that required reoperation and early graft dysfunction (defined in part by a PaO2/FiO2 ratio less than 150 during the first 24 hours) were the variables that best predicted death in the early postoperative period following lung transplantation. PMID- 12890404 TI - [Respiratory muscle disorders in chest wall diseases]. PMID- 12890405 TI - [Dr. Antonio Corbella Fondebila's treatise on chest diseases]. PMID- 12890406 TI - [Chronic thromboembolic pulmonary hypertension associated with endomyocardial fibrosis of the right ventricle]. AB - Chronic thromboembolic pulmonary hypertension is a rare sequela to an acute untreated or recurrent pulmonary embolism. The mechanisms that underlie the failure to resolve the thrombus are still uncertain. As most patients are not diagnosed until a relatively late stage, little is known about the course of their illness. We report the case of a 51-year-old woman who had previously been diagnosed with and operated on for endomyocardial fibrosis of the right ventricle and who developed chronic thromboembolic pulmonary hypertension several years later. PMID- 12890407 TI - [Stevens-Johnson syndrome associated with atypical pneumonia]. AB - Stevens-Johnson syndrome is characterized by generalized exanthema associated with high fever, catarrhal symptoms and mucositis. Various etiologies have been implicated, particularly numerous medications and certain agents of atypical pneumonia. Stevens-Johnson syndrome leads to death in up to 5% of cases. We describe the case of a 30-year-old woman with bilateral atypical pneumonia accompanied by severe generalized exanthema that required hospitalization in the serious burns unit of our hospital. She was diagnosed with Stevens-Johnson syndrome following atypical pneumonia caused by Mycoplasma pneumoniae with microbiological and pathological confirmation. PMID- 12890408 TI - [Disseminated tuberculosis due to Mycobacterium africanum]. PMID- 12890409 TI - [One name, one disease: chronic obstructive pulmonary disease]. PMID- 12890410 TI - [Chronic obstructive pulmonary disease in 2002]. PMID- 12890411 TI - Asthma: will the therapeutic strategy change? PMID- 12890412 TI - Is Helicobacter pylori infection associated with chronic idiopathic urticaria? AB - BACKGROUND: Chronic idiopathic urticaria (CIU) is one of the most frequent skin diseases, however its causes remain unknown in the vast majority of cases. There is increasing evidence for systemic effects of gastric Helicobacter pylori infection, which may result in extra gastrointestinal disorders. Although CIU can result from several causes, a possible relationship between chronic urticaria and Helicobacter pylori has been recently suggested. OBJECTIVE: The aim of this study is to determine the prevalence of Helicobacter pylori infection in a series of patients with CIU, and measure the effectiveness of eradication therapy on the skin disease. METHOD: We assess Helicobacter pylori infection by 13C urea breath test (UBT) in 21 CIU patients. Amoxicillin, clarithromycin, and omeprazole were given to infected patients for seven days. The results of therapy were assessed by urea breath test one month after therapy. Urticaria and gastrointestinal symptoms were assessed on enrolment and for six months after eradication. RESULTS: Prevalence of Helicobacter pylori infection was of 71.4 % (15/21). There were no differences concerning age (31.83 vs. 33.82 years) and duration of symptoms (38.40 vs. 35.38 months) in patients with respectively positive and negative UBT. Helicobacter pylori eradication rate was of 80 % (12/14). Three patients had clinical improvement with total resolution of urticaria starting immediately after eradication therapy, being able to completely withdrawn antihistamine and corticosteroids therapy without complaints. In relation to the rest of the patients, these three had smaller duration of urticaria disease (14.3 vs. 43.7 months; p = 0.038) and greater titters in the UBT results (42.39 vs. 25.81; p = 0.073). DISCUSSION: Although some authors found convincing evidence of the involvement of Helicobacter pylori as one possible cause of chronic urticaria, our results have failed to confirm the existence of this etiological association. The most remarkable finding was that those patients who had clinical remission of disease were the ones with greater UBT titters suggesting a role for the amount of colonization by Hp in the pathogenesis of urticaria disease. PMID- 12890413 TI - [Hypersensitivity pneumonitis induced in guinea pigs by antigens to bat feces]. AB - A bat feces extract was able to induce the synthesis of specific IgG in rabbits and IgE in atopics. These findings were corroborated by skin tests, RAST and RAST inhibition. Nowadays, data concerning the experimental induction of hypersensitivity pneumonitis using a glycoprotein of bat feces are presented. This antigen was aerosolized for inhalation by adult guinea pigs for 12 weeks to detect specific serum IgM, IgG and IgE antibodies as well as sensitized T-CD4 cells. Histopathological studies of the lungs showed interstitial infiltrates of macrophages and lymphocytes, cellular bronchiolitis and single non-necrotizing granulomas from the seventh to the ninth weeks. From the tenth week to the end of the experiment the lesions of the lungs progressively worsened. The results from this animal model suggest that the chronic contact with the bat feces'antigen can induce other inflammatory lung reactions than those IgE dependant. PMID- 12890414 TI - Effects of specific immunotherapy on the development of new sensitisations in monosensitised patients. AB - BACKGROUND: Specific immunotherapy (SIT) is the only treatment that interferes with the basic pathophysiological mechanisms of allergic disease and is widely used in the management of clinically significant respiratory IgE-mediated diseases. Nevertheless, until recently, information on the influence of SIT on the development of new allergic sensitisations has been scant. METHODS: One hundred consecutive patients (45 males and 55 females, aged 6 to 69 years) with respiratory allergic diseases and attending the allergy unit of a general hospital were selected. All had been diagnosed by clinical history and skin prick tests of allergic rhinitis and/or asthma, were monosensitised (71 to Dermatophagoides spp, 22 to Parietaria judaica pollen and 7 to grass pollen) and had been followed up as outpatients between 1990-98. Sixty-six patients had been treated with conventional SIT for at least 3 years, while thirty-four followed only environmental measures and drug treatment. Family atopy status (first-degree relatives), smoking, family pets (cat and/or dog), rhinitis and/or asthma symptom score and inhalant skin prick tests to the same aeroallergens were compared between baseline and after 3 to 5 years of treatment. RESULTS: No statistically significant differences in the development of new sensitisations were observed between the two groups (36.4 % of SIT-treated patients versus 38.2 % in control group, RR = 0.97, CI 95 %: 0.72-1.3). Smoking, family atopy history and pets did not appear to be risk factors for the development of neosensitisations (p < 0.05). Nevertheless, SIT-treated patients presented a better clinical score than the control group, with improvements of 89.4 % and 61.8 %, respectively (p = 0.007). CONCLUSIONS: Three-year SIT did not protect against development of new sensitisations in monosensitised allergic rhinitis or asthma. Smoking, family atopy history and pets were not associated with development of new sensitisations. Clinical score improved significantly in the SIT-treated group compared with drug-treated patients. PMID- 12890415 TI - Retrospective evaluation of epidermal skin prick tests in patients living in Aegean region. AB - Sensitization to aeroallergens of Aegean region is not well decumented. In this study we evaluated the epidermal skin prick test results of the patients who applied to allergy outpatient department retrospectively. Epidermal skin prick test of the 5055 patients were evaluated. Of these patients 2638 (52 %) were female, 2417 (48 %) male, 1213 (24 %) adult and 3842 (76 %) pediatric patient, 1163 (23 %) patients were allergic rhinitis, 2477 (49 %) were bronchial asthma, 505 (10 %) were allergic rhinitis with bronchial asthma, 556 (11 %) were chronic urticaria, 253 (5 %) were wheezy infant and 101 (2 %) patients were atopic dermatitis, 2932 (58 %) had atopy history in their first and second degree relatives. Patients were aged between 3.5 months and 79 years (mean 14.1 3.2 years and median 11 years). In epidermal skin prick tests sensitization to house dust mites (D. farinae, D. pteroniyssinus), pollens (grass, cereals and trees), moulds, animal danders, foods (especially in early childhood) and cockroach were evaluated. Our data indicate that allergens that may be the cause of the high prevalence of allergic diseases in Izmir are probably produced by pollens and mites. PMID- 12890416 TI - Asthma camp. Quality of life questionnaires. AB - Asthma is one of the most common chronic chilhood disease. Asthmatic children need a self-management educational program, besides the medical treatment and clinical control. Camps are the ideal place to complete the educational program, to get the right control of the disease and to build the child confidence. Madrid Castilla la Mancha Allergy Society organizes every year a summer camp in a town, near Madrid. Fifty-four children stay for one week with certified allergists, fellows-in-training, nurses and cheer-leaders.Fifty-four children from 8 to 14 years old, with moderate to severe asthma participate in the camp. The camp includes educational, athletic and social activities. Campers participate daily in instructive and didactic teaching sessions. We evaluate quality of life with a questionaire filled in the camp and two months later. Results are analysed into two age groups, one between 8-10 years old and the other from 11 to 14 years old. The first group get a good management and improve their vision of the disease, quite the contrary, adolescents face up to a chronic disease with daily treatment, feeling asthma as a restriction of their autonomy. Even so, they identify crisis and learn to control them in an special risky life period. Asthma camps is a good experience for both groups and also for sanitary equipment. PMID- 12890417 TI - PFAPA syndrome: with regard to a case. AB - BACKGROUND: PFAPA syndrome (Periodic Fever, Aphtas, Pharyngitis and cervical Adenopathies) is one of the causes of periodic fever in pediatrics and it is characterised by high fever, pharyngitis, cervical adenitis and aphtous stomatitis. Its etiopathogeny is unknown. The diagnosis is clinical and the findings of laboratory are unspecified. One or two doses (1 mg/kg) of oral prednisone are enough for a fast resolution of the clinic. It is a benign syndrome and no sequels have been noticed after its disappearance, usually in four years from its beginning. CLINICAL CASE: We present the case of a 10-year old patient who has been diagnosed of PFAPA syndrome after 3 years and a half of characteristic clinical bouts, with the fulfilment of diagnostic criteria and after having excluded other entities of similar presentation. CONCLUSIONS: Periodic episodes of high fever, pharyngitis and cervical adenitis with a bad response to the conventional treatment should alert us to the PFAPA syndrome. The recognition of this entity will help us to improve the diagnostic and therapeutical focusing, lowering also the anxiety that these cases produce. PMID- 12890418 TI - [Exercise-induced anaphylaxis after apple intake]. AB - Exercise-induced anaphylaxis frequently is related to food ingestion, so that it receives the name of "food-dependent exercise-induced anaphylaxis". The food identification is difficult in some patients. We report two patients with apple dependent exercise-induced anaphylaxis. STUDY: hematimetries, biochemistries, VMA and 5-HIAA in urine. Complement levels, antinuclear antibodies and viruses serology. ALLERGOLOGIC STUDY: skin prick test with a common inhalant. Skin prick test and prick by prick with different foods. Total Ig E level and specific Ig E determination by the CAP-Pharmacia System. Oral challenge test with apple, exercise test and exercise challenge test after eating an apple. Prick by prick test with fresh apple was positive. Specific Ig E determination by the CAP Pharmacia System revealed a positive result. We get to reproduce the episode with an exercise challenge test after eating an apple in the same conditions of usual exercise in both patients. PMID- 12890419 TI - [Sublingual immunotherapy in children. Immunotherapy Committee of the Spanish Society for Clinical Immunology and Pediatric Allergology]. AB - Sublingual immunotherapy is currently attracting growing interest because of its ease of administration and, according to previous studies, its infrequent and mild adverse effects. However, at least in children, the efficacy of this therapy has not been completely demonstrated. In addition, the mechanisms of action remain to be elucidated since few studies have been published and the results have been contradictory and sometimes inconclusive. For this reason, we performed a literature review through the MEDLINE database, selecting double-blind studies carried out in children. Only 10 studies meeting these requirements were retrieved. All the studies were performed by European researchers and nine were published in European journals. Efficacy was evaluated by clinical parameters and by reduction in medication use. The results on efficacy are not homogeneous, although most support the utility of this route of administration. Moreover, reports of allergens other than those used in these studies dust mites and grass pollens are lacking. In conclusion, further studies evaluating the efficacy of this therapy in children are required. Among the general population, if the efficacy of sublingual immunotherapy in the treatment of sensitization to hymenoptera venoms were demonstrated, as has been the case with subcutaneous immunotherapy, the utility of this route of administration would be definitively confirmed. Finally, sublingual immunotherapy could be used in children who have shown systemic reactions to subcutaneous immunotherapy or who refuse to undergo injections. PMID- 12890421 TI - Risk characterisation of chemicals in food and diet. PMID- 12890422 TI - Anti-inflammatory properties of pro-inflammatory interferon-gamma. AB - Production of interferon-gamma (IFNgamma) in response to infection is a hallmark of innate and adaptive immunity. In addition to the pivotal role of IFNgamma in host defense, its excessive release has been associated with the pathogenesis of chronic inflammatory and autoimmune diseases. In fact, knockout models reveal that IFNgamma plays a key role in mediating a number of pathological processes related to chronic immune activation. On the other hand, evidence has been accumulated in recent years that supports the concept of a dual role of IFNgamma in inflammation. Here, we review anti-inflammatory aspects of IFNgamma in the regulatory network of cytokine biology. These include induction of anti inflammatory molecules such as interleukin (IL)-1 receptor antagonist (IL-1Ra) and IL-18 binding protein (IL-18BP), modulation of pro-inflammatory cytokine production, activation of apoptosis, and interference with the signal transduction machinery by induction of suppressors of cytokine signaling (SOCS). PMID- 12890423 TI - Effect of montelukast on polymorphonuclear leukocyte functions in asthmatic patients. AB - Leukotriene receptor antagonists are being used widely in the treatment of bronchial asthma. They have been shown to possess anti-inflammatory properties, but there is no sufficient data about their effects on polymorphonuclear leukocyte functions. The aim of this study was to investigate the effects of montelukast, a specific cysteinyl leukotriene-1 receptor antagonist, on human polymorphonuclear leukocyte (PMN) functions (phagocytic and intracellular killing activity) in asthmatic patients. Fifteen mild to moderate asthmatic patients were included in the study. They were treated with montelukast (10 mg/day per os) in addition to their previous medications for 2 weeks. Whole blood samples of patients were taken before and after this treatment period. Phagocytic activities and intracellular killing activities of polymorphonuclear leukocytes isolated from whole blood samples were tested by using appropriate technics. Phagocytic and intracellular killing activities of PMNs were significantly increased (p<0.001, p<0.05) by montelukast compared to those before treatment. These results show that montelukast has an enhancing effect on PMN functions in asthmatic patients. PMID- 12890424 TI - Differential effects of synthetic indoloquinolizines on in vitro rat lymphocyte and macrophage functions. AB - Indoloquinolizines are natural alkaloid indole products grouped as beta carbolines. These compounds are commonly associated with neurological activities, but little is known about their role as immunomodulating agents. The present study was undertaken to evaluate the effects of synthetic indoloquinolizines on in vitro parameters of rat lymphocyte and macrophage functions. It was observed that proliferation of thymic lymphocytes was significantly (p<0.05) increased (20 30% increase) by dihydro-indoloquinolizinium chloride (2). dihydro indoloquinolizinyl-ethanone (3). and dimeric dihydro-indoloquinolizinium dichloride (6). whereas dimeric indoloquinolizine (7). caused up to 40% increase in lymphoproliferation at concentrations ranging from 10(-11) to 10(-5) M, compared with untreated control. In contrast, indoloquinolizinium chloride (4) and indoloquinolizine (5). were toxic for lymphocytes at concentrations from 10( 9) to 10(-5) M, and compounds 6 and 7 were toxic at 10(-5) M. In addition, nitric oxide production by LPS-treated peritoneal macrophages was significantly (p<0.05) increased (up to 30% increase) by compounds 4 and 5 at concentrations of 10(-11) to 10(-5) M, and 10(-5) M, respectively; however, compounds 6 and 7 were toxic for macrophages at all concentrations tested. Furthermore, TNF-alpha production was also significantly increased (p<0.01) by compounds 4 and 5 (up to 30-fold increase) compared with untreated control. These novel synthetic indoloquinolizines could serve as immunotherapeutic agents by selectively increasing the pool of activated T lymphocytes or stimulating macrophage functions, with potential use in the treatment of infectious diseases including AIDS and cancer. PMID- 12890425 TI - Inhibition of induction of myofibroblasts by interferon gamma in a human fibroblast cell line. AB - Interferon gamma (IFNgamma) has been reported as a possible therapeutic agent for contractile diseases in clinical trials and in vitro studies. It is not yet clear, however, whether IFNgamma simply inhibits myofibroblast generation or downregulates alpha smooth muscle actin (alphaSMA) production in myofibroblasts. In this study, we attempted to clarify how IFNgamma acts in the generation of myofibroblasts, and the production of alphaSMA by myofibroblasts, using immunofluorescence staining, cell capture enzyme immunoassay (CC-EIA) and the reverse transcription polymerase chain reaction (RT-PCR) for alphaSMA. We examined whether IFNgamma could block the TGFbeta1-promoted changes in myofibroblasts or the generation of myofibroblasts by TGFbeta1. IFNgamma strongly blocked the generation of myofibroblasts and moderately inhibited the production of alphaSMA in TGFbeta1-promoted myofibrobasts. These findings indicate that IFNgamma may be effective in the early stage of contractile diseases to prevent the progression of contractile lesions. PMID- 12890426 TI - Proteoglycan isolated from Phellinus linteus activates murine B lymphocytes via protein kinase C and protein tyrosine kinase. AB - Medicinal mushrooms are increasingly used to treat a wide variety of disease processes. Aqueous extract from the fruiting body or mycelia of Phellinus linteus has been reported to produce antitumor and immunomodulatory activities in vivo and in vitro. However, the therapeutic mechanism has not been known. In the present study, we investigated whether proteoglycan (PL) isolated from P. linteus has an effect on the immunomodulatory activities of the murine splenic lymphocytes (MSLs). Treatment with PL caused a four-fold augmentation in [3H]thymidine incorporation compared to untreated control group in MSLs. Flow cytometric analysis indicated that the affected cell population was mainly CD19(+) cells, but not CD3(+) cells. These data suggested that the main target of PL was the B cells, but not T cells. PL also enhanced the expression of co stimulatory molecules, CD80 and CD86, in murine B cells in a time-dependent manner. Accordingly, we investigated if intracellular [Ca(2+)] and reactive oxygen intermediates (ROI) were the principal downstream components that contributed to PL-induced activation, with respect to both increases of proliferation and induction of co-stimulatory molecules. However, PL has no influence on the [Ca(2+)] concentration and the ROI formation in murine B cells, whereas the genistein, protein tyrosine kinase (PTK) inhibitor or staurosporine, protein kinase C (PKC), blocked the proliferation and the induction of co stimulatory molecules, CD80 and CD86, in B cells stimulated with PL. Taken together, these data suggest that PL is a biological response modifier that stimulates proliferation and expression of co-stimulatory molecules in B cells, probably by regulating PTK and PKC signaling pathways. PMID- 12890427 TI - In vitro anti-inflammatory activity of Phlebodium decumanum. Modulation of tumor necrosis factor and soluble TNF receptors. AB - The immunomodulatory activity of a standardized water soluble fraction of the fern Phlebodium decumanum (EXPLY-37) previously shown to have "in vivo" anti inflammatory activity was analyzed "in vitro". This extract inhibited tumor necrosis factor (TNF) production by macrophages activated with lipopolysaccharide (LPS) or LPS plus interferon (IFN)-gamma. In contrast, nitric oxide (NO) and interleukin (IL)-1beta production were not affected in the same cultures, whereas IL-6 production was partially inhibited. More interestingly, EXPLY-37 increased the release of soluble TNF-receptor 2 (sTNFR2) and of IL-1R antagonist (IL-1Ra) but not of sTNFR1, by activated macrophages. EXPLY-37 had no effect on T lymphocyte activation, measured as proliferation as well as expression of early and late cell surface antigens CD69, CD25 (IL-2R-alpha) and CD71 (transferrin receptor) at the cell membrane. At the molecular level, EXPLY-37 did not inhibit the activation of the nuclear factor kappa B (NF-kappaB) transcription factor by TNF. In summary, EXPLY-37 has two anti-inflammatory activities "in vitro": it decreases TNF production and increases IL-1Ra and sTNFR2, which may be able to neutralize IL-1 and TNF activity, respectively. PMID- 12890428 TI - Toll-like receptor-mediated activation of B cells and macrophages by polysaccharide isolated from cell culture of Acanthopanax senticosus. AB - We investigated the mechanism of the immunomodulatory action of polysaccharide (ASP) isolated from a cell culture of Acanthopanax senticosus. ASP was found to directly increase the proliferation and differentiation of B cells, and the cytokine production of macrophage, but not the proliferation and cytokine production of T cells. Since ASP cannot penetrate the cell membrane due to its large molecular mass, such cellular activation may be caused by the surface binding of ASP to receptors expressed on B cells and macrophages. The possibility that TLRs, which are known to be involved in immune-related responses, may be the receptor(s) of ASP was investigated. The immunomodulating activities of ASP on the B cells and macrophages of C3H/HeJ mice, expressing a defective toll-like receptor (TLR)-4, were decreased versus the corresponding cells from C3H/HeN mice. In addition, the activities of ASP on B cells and macrophages were significantly reduced by treating the cells with antibodies to TLR4 and TLR2 prior to ASP, suggesting that both of them are the possible receptors of ASP. The ligation of TLRs induced by ASP was able to activate mitogen-activated protein kinases (MAPKs), such as Erk1/2, p38 and JNK, and the transcription factor NF kappaB. Although ASP was shown to activate the TLR signaling cascades in the same manner as lipopolysaccharide (LPS), these two could be differentiated by the finding that polymyxin B (PMB), a specific inhibitor of LPS, did not significantly affect the activities of ASP on B cells and macrophages. Taken together, our results demonstrate that ASP, isolated from a cell culture of A. senticosus, activates B cells and macrophages by interacting with TLRs and leading to the subsequent activation of mitogen-activated protein kinases and NF kappaB. PMID- 12890429 TI - Oral administration of royal jelly inhibits the development of atopic dermatitis like skin lesions in NC/Nga mice. AB - We have shown previously that in addition to IL-4, IL-5 and IL-10, antigen specific interferon-gamma (IFN-gamma) production by spleen cells from ovalbumin (OVA)/Alum-immunized mice is inhibited by the administration of royal jelly (RJ). Since it has been shown that both Th1 and Th2 cytokines play pathogenic roles in the generation of atopic dermatitis (AD), we have examined whether RJ suppresses the development of AD-like skin lesions in NC/Nga mice induced by repeated application of picryl chloride (PiCl) under specific pathogen-free (SPF) conditions. Oral administration of RJ to the PiCl-treated NC/Nga mice inhibited the development of AD-like skin lesions in these mice as exemplified by the significant decrease in the total skin severity scores and the decrease in hypertrophy, hyperkeratosis, and infiltration of the epidermis and corium by inflammatory cells. IFN-gamma production by spleen cells from PiCl-treated NC/Nga mice in response to TNP-KLH was partially but significantly inhibited by the oral administration of RJ, while IFN-gamma production by Con A-stimulated spleen cells was not affected. Since inducible nitric oxide (NO) synthase (iNOS)-derived NO has been suggested as an important immunoregulatory mediator in inflammatory autoimmune diseases, we have also examined the expression of iNOS in the dorsal skin lesions of PiCl-treated NC/Nga mice. Interestingly, the expression of iNOS was significantly increased in the skin lesions of RJ-administered mice compared with those of control PBS-administered mice. Thus, our results suggest that RJ suppresses the development of AD-like skin lesions in PiCl-treated NC/Nga mice, possibly by a combination of down-regulating TNP-specific IFN-gamma production and up-regulating iNOS expression. PMID- 12890430 TI - Comparison of the efficacy of IGIV-C, 10% (caprylate/chromatography) and IGIV-SD, 10% as replacement therapy in primary immune deficiency. A randomized double blind trial. AB - A novel method of large-scale chromatography has been developed to improve recovery and purity of immunoglobulin G (IgG) from pooled plasma. The current study compares safety, toxicity and efficacy of two intravenous immunoglobulin products: a novel formulation, IGIV caprylate/chromatography (IGIV-C; Gamunex, 10%) and a licensed solvent/detergent-treated product, Gamimune N, 10% (IGIV-SD). The study, a randomized, double-blind, parallel group, therapeutic equivalence trial, was conducted at 25 treatment centers in Canada and the United States. Patients (n=172) having confirmed chronic primary immunodeficiency (PID), aged 1 75 years, and receiving IGIV therapy were enrolled. For 9 months, patients were treated with IGIV-C or IGIV-SD in accordance with the patient's individualized treatment regimen utilized before study entry. The primary endpoint was the proportion of patients with >or=1 validated acute sinopulmonary infection during the treatment period. Secondary endpoints included the proportion of patients with all infections, time to first infection, annual infection rates, lung function parameters, infusion-related safety and viral safety. The annual validated infection rate in the IGIV-C group was 0.18 compared to 0.43 in the IGIV-SD group (p=0.023). Nine patients receiving IGIV-C experienced validated infections, compared to 17 patients in IGIV-SD group (p=0.06). Acute sinusitis (validated plus clinically defined) was less frequent in the IGIV-C group (p=0.012). Presence of bronchiectasis did not affect efficacy. Adverse reactions were similar in frequency and severity in both groups. No evidence of viral transmission was observed. IGIV-C appears to be superior to IGIV-SD in preventing validated sinopulmonary infections, especially acute sinusitis, in patients with PID. PMID- 12890431 TI - Nafamostat mesilate suppresses NF-kappaB activation and NO overproduction in LPS treated macrophages. AB - Nafamostat mesilate (NM), a clinically used serine protease inhibitor, suppressed the overproduction of nitric oxide (NO) and the expression of inducible nitric oxide synthase (iNOS) in RAW264.7 murine macrophages treated with lipopolysaccharide (LPS, 100 ng/ml); however, it had little effect on endothelial NOS (eNOS) in human umbilical vein endothelial cells (HUVEC). Electrophoretic mobility shift assay (EMSA) revealed that LPS activated nuclear factor-kappaB (NF kappaB) in RAW264.7 cells and that this activation was suppressed by nafamostat mesilate. Western blotting showed that nafamostat mesilate suppressed the phosphorylation and degradation of inhibitor kappaB-alpha (IkappaB-alpha), which holds NF-kappaB in the cytoplasm in an inactivated state. Our observations suggest that nafamostat mesilate is a candidate agent for various diseases such as ischemia-reperfusion, graft rejection, inflammatory diseases, and autoimmune diseases, in which iNOS and/or NF-kappaB are upregulated. PMID- 12890433 TI - Bridging the gap across fields of practice. PMID- 12890432 TI - Serotonin, serotonin 5-HT(1A) receptors and dopamine in blood peripheral lymphocytes of major depression patients. AB - There are increasing evidences of cell markers present in the immune and the nervous systems. These include neurotransmitter receptors and transporters. Serotonin receptor subtypes are related to depression and also have been shown to be present in certain cells of the immune system. In the present report, we determined the presence of 5-HT(1A) receptors by the binding of the selective agonist 8-hydroxy-2-(di-n-propyl-amino)tetralin in lymphocytes of peripheral blood isolated by Ficoll/Hypaque gradients from controls and depressed patients. The capacity of these receptors was around 24 fmol/10(6) cells in both groups of subjects, without significant difference among them. The affinity was in the nM range and either differ between controls and patients. Serotonin, 5 hydroxyindoleacetic acid, dopamine and 3,4-dihydroxyphenylacetic acid were determined by HPLC with electrochemical detector. There were no significant differences between controls and major depression patients in the values obtained for rich and poor platelet plasma or in the isolated cells. However, there was a reduction in serotonin turnover rate indicated by an increase in the ratio serotonin/5-hydroxyindoleacetic acid, but not in that of dopamine, in lymphocytes of major depression patients. Thus, there is a serotonergic dysfunction in immune circulating cells of major depression patients, without changes in the number of 5-HT(1A) receptors, although the coupling of these receptors to transduction mechanisms could be affected and may be related to the alteration of 5-HT turnover rate. PMID- 12890434 TI - Lateral epicondylalgia: a musculoskeletal physiotherapy perspective. AB - Tennis elbow or lateral epicondylalgia (LE) is a challenging musculoskeletal condition to treat. This is largely due to the lack of research-based evidence of the clinical efficacy of the myriad of treatment approaches espoused in the literature. In view of this, successful rehabilitation of LE is based on choosing treatments that address the physical impairments found during clinical examination. The primary physical impairment in LE is a deficit in grip strength predominately due to pain and its consequences on motor function. Hence the mainstay of successful management of this condition is therapeutic exercise, providing it is not pain provocative. Adjunctive procedures such as manipulative therapy and sports taping techniques have recently been shown to provide substantial initial pain relief. Early relief of pain in the rehabilitation program helps accelerate recovery and most importantly motivates the client to persist with the therapeutic exercise program. The manipulative therapy and taping treatments presented in this masterclass warrant consideration in the clinical best practice management of LE, and serve as a model for other similar musculoskeletal conditions. PMID- 12890435 TI - A systematic review of physiotherapy for spondylolysis and spondylolisthesis. AB - The purpose of this systematic review was to assess the evidence concerning the effectiveness of physiotherapy intervention in the treatment of low back pain related to spondylolysis and spondylolisthesis. A literature search of published and unpublished articles resulted in the retrieval of 71 potential studies on the subject area. Fifty-two of the 71 articles were studies, and these studies were reviewed using preset relevance criteria. Given the inclusion and exclusion criteria chosen for this systematic review, there were very few acceptable studies and only two studies met the relevance criteria for the critical appraisal. Both studies provide evidence to suggest that specific exercise interventions, alone or in combination with other treatments, have a positive effect on low-back pain due to spondylolysis and spondylolisthesis; however, the type of exercise used was different in the two studies. In this review, very few prospective studies were found that examined the efficacy of physiotherapy on the topic area; therefore, few conclusions can be made, and further research is warranted. PMID- 12890436 TI - Relationship between cranio-cervical flexion range of motion and pressure change during the cranio-cervical flexion test. AB - The purpose of this study was to quantify the sagittal angular displacement of the head (cranio-cervical flexion) for the five incremental stages of the cranio cervical flexion test (CCFT). Range of cranio-cervical flexion during the CCFT was measured using a digital imaging method in 20 healthy volunteer subjects. The intra-and inter-rater reliability of the digital imaging technique for the assessment of this movement were also examined. The results of this study demonstrated a linear relationship between the incremental pressure targets of the CCFT and the percentages of full range cranio-cervical flexion range of motion (ROM) measured in the supine lying position of the test using a digital imaging technique. A mean of 22.9% full range cranio-cervical flexion was used to reach the first pressure target of the CCFT followed by linear increments up to 76.6% for the last stage of the test. An increasing amount of cranio-cervical flexion ROM was used to achieve the five successive stages of the CCFT reflecting an increasing contractile demand on the deep cervical flexor muscles. Excellent inter-rater (ICC=0.994) and intra-rater reliability (ICC=0.988-0.998) were demonstrated for the angular measurements using this digital imaging technique. PMID- 12890437 TI - Quantitative analysis of traction in the glenohumeral joint. In vivo radiographic measurements. AB - PURPOSE: To analyse change in distance between the humeral head and the glenoid fossa during traction in the maximally loose-packed position (MLPP) and the maximally closed-packed position (MCPP) under standardized conditions. SUBJECTS: Six healthy subjects (three male and three female) with a mean age of 40.5 years, volunteered to participate in this study. MATERIALS AND METHODS: Subjects were placed with the right shoulder in a modified shoulder brace (Otto Bock Armabduktions-Orthese in Modular Bauweise in 45 degrees abduction in the plane of the scapula with neutral rotation (MLPP). A standard anterior-posterior radiograph of the glenohumeral joint was made. A 14 kg traction force was applied for 40s, and a second radiograph was made. The same procedure was repeated with the shoulder placed in the MCPP, which was 90 degrees abduction and 90 degrees external rotation. A radiologist, blinded for the variable traction or no traction, performed all radiographic measurements. Measurements were made on the same radiographs on two separate occasions (O1 and O2) with a 2-month interval. RESULTS: No significant differences were found in mean distance between the humeral head and the glenoid fossa during traction in the MLPP compared to traction in the MCPP (O1: P=1.00) and (O2: P=0.63). CONCLUSIONS: Application of a 14 kg force does not result in a significant increase of distance between the humeral head and the glenoid fossa. No significant difference was found between the change in distance of the humeral head and the glenoid fossa after traction in the MLPP compared to traction in the MCPP. PMID- 12890438 TI - The effect of cervical rotation on blood flow in the contralateral vertebral artery. AB - Twenty asymptomatic volunteers (mean age 33 years, range 26 - 54 years) underwent investigation using duplex Doppler ultrasound with real-time imaging and colour flow enhancement. With the subjects seated, peak velocity at C1-2 and volume flow rate at C5-6 were measured in the artery contralateral to the direction of rotation, in the four positions of neutral, 45 degrees and end range rotation, plus a subsequent neutral position. No change in peak velocity at C1-2 between the initial neutral measurement and the measurements at 45 degrees and end range rotation was found (P>0.05). Peak velocity was less in both vertebral arteries on return to the neutral position as compared with end range rotation, however the difference was significant for the left vertebral artery only (P=0.005). This lends support for the rest period, which is taken between cervical movement tests when conducting pre-manipulative testing, to allow for any latent effect on blood flow of the tests themselves. There was no change in volume flow rate between any of the test positions (P=0.349). There was no indication of a cumulative effect of the test procedure (P>0.05). PMID- 12890440 TI - Validating clinical reasoning: a question of perspective, but whose perspective? AB - This paper explores the notion of validity from different perspectives and addresses its integration with clinical reasoning. Whilst valuing the evidence based practice (EBP) perspective in our quest for validation of practice, the authors posit that other perspectives may provide added value through their potential for offering validity in differing circumstances. The reality of the EBP aspiration is discussed and nuances within the evolved term 'research enhanced practice' are shared. Validity theory applied through different perspectives may, it is argued help to get to grips with the balancing act of validating manual therapy practice. PMID- 12890439 TI - Adherence to rehabilitation in patients with low back pain. AB - This study investigated adherence to low back pain rehabilitation in the clinical setting. Adherence was assessed in 105 (71 male and 34 female) patients attending private physiotherapy clinics for rehabilitation of low back pain. Three aspects of adherence were measured over the 4-week study period: attendance at clinic based rehabilitation sessions, adherence to a home exercise component of rehabilitation, and adherence to activities and advice during clinic-based rehabilitation. Rehabilitation outcome was measured using the Patients' Measure of Perceived Rehabilitation (McDonald & Hardy 1990) and the Physiotherapists' Measure of Perceived Rehabilitation (McDonald & Hardy 1990). It was found that patients attended 87.7% of their scheduled physiotherapy rehabilitation appointments and reported completing 71.6% of their prescribed home exercises. In relation to adherence to clinic-based rehabilitation activities (as measured by the Sport Injury Rehabilitation Adherence Scale, Brewer et al. 2000), patients scored an average of 11.6/15. Although no gender differences were found, compensable patients adhered significantly less to clinic-based rehabilitation activities than did their non-compensable counterparts. Further, it was found that higher levels of adherence to clinic-based activities significantly predicted both the patients' and physiotherapists' perception of degree of rehabilitation at the end of the 4-week rehabilitation period. These findings are discussed in relation to rehabilitation strategies for physiotherapists. PMID- 12890441 TI - False-negative extension/rotation pre-manipulative screening test on a patient with an atretic and hypoplastic vertebral artery. PMID- 12890442 TI - The morphometric relationship of Upper Cave 101 and 103 to modern Homo sapiens. AB - Since the discovery of the Homo sapiens crania from the Upper Cave of Zhoukoudian in northern China (UC 101, UC 102, and UC 103), no clear consensus has arisen regarding their affinities with modern populations. We use linear craniofacial measurements to compare UC 101 and UC 103 to a worldwide sample of H. sapiens that includes Paleoamericans and Archaic Indians, and employ Mahalanobis distance analysis and associated unweighted, unrestricted canonical variate analysis for the comparisons. Analyses indicate that UC 101 has consistent affinities with Easter Island and European populations, whereas UC 103 has more tenuous similarities with Australo-Melanesian groups. Both fossils exhibit some similarities to certain Paleoamerican and Archaic Indian individuals, but rarely cluster together. Upper Cave 103 is more of an outlier to modern populations than is UC 101. The fossils are not representative of any group to which they have been compared, but may be part of the generalized population that was ancestral to Paleoamericans. PMID- 12890443 TI - Early Holocene human skeletal remains from Santana do Riacho, Brazil: implications for the settlement of the New World. AB - In this study we compare the cranial morphology of several late Paleoindian skeletons uncovered at Santana do Riacho, Central Brazil, with worldwide human cranial variation. Mahalanobis Distance and Principal Component Analysis are used to explore the extra-continental morphological affinities of the Brazilian Paleoindian sample. Santana do Riacho is a late Paleoindian burial site where approximately 40 individuals were recovered in varying states of preservation. The site is located at Lagoa Santa/Serra do Cipo, State of Minas Gerais. The first human activities in this rockshelter date back to the terminal Pleistocene, but the burials are bracketed between circa 8200 and 9500BP. The collection contains only six skulls well-enough preserved to be measured. The Santana do Riacho late Paleoindians present a cranial morphology characterized by long and narrow neurocrania, low and narrow faces, with low nasal apertures and orbits. The multivariate analyses show that they exhibit strong morphological affinities with present day Australians and Africans, showing no resemblance to recent Northern Asians and Native Americans. These findings confirm our long held opinion that the settlement of the Americas was more complicated in terms of biological input than has been widely assumed. The working hypothesis is that two very distinct populations entered the New World by the end of the Pleistocene, and that the transition between the cranial morphology of the Paleoindians and the morphology of later Native Americans, which occurred around 8-9ka, was abrupt. This, in our opinion, is a more parsimonious explanation for the diversity detected than a long, local microevolutionary process mediated by selection and drift. The similarities of the first South Americans with sub Saharan Africans may result from the fact that the non-Mongoloid Southeast Asian ancestral population came, ultimately, from Africa, with no major modification in the original cranial bau plan of the first modern humans. PMID- 12890444 TI - The inverse relationship between species diversity and body mass: do primates play by the "rules"? AB - Evolutionary biologists have long commented on a seemingly universal "rule" of nature-that in large taxonomic assemblages from groups as diverse as bacteria, plants, insects, marine invertebrates, fish, reptiles, amphibians, birds, and mammals, there exists a frequency distribution of body sizes among species that is highly skewed to the right (positive skewness). This distribution reflects the strong inverse, or negative, relationship often noted between mean body size of taxa and the number of species they contain--i.e., the observation that small body size is often associated with high species diversity (speciosity). This is sometimes "explained" by recourse to the idea that smaller-bodied taxa are able to subdivide their environments more finely than larger-bodied taxa. With but few exceptions, the applicability of this "rule" to the Order Primates has not been studied in any detail. In this study I address the following questions of (paleo)anthropological interest: (1) How speciose is the Order Primates? (2) Does this biological "rule" characterize the Order Primates (at any taxonomic level) in any meaningful way? (3) Does the association between speciosity and body mass within the Order Primates provide any useful models for interpreting and/or predicting speciosity in the fossil primate record? Using phylogenetically independent contrasts methods, I conclude that the answers to those three questions are: (1) not very; (2) no; and (3) not particularly (with the possible exception of larger-bodied taxa). PMID- 12890445 TI - Functional and phylogenetic implications of molar flare variation in Miocene hominoids. AB - Comparative analyses of molar shape figure prominently in Miocene hominoid evolutionary studies, and incomplete understanding of functional and phylogenetic influences on molar shape variation can have direct consequences for the interpretation of fossil taxa. Molar flare is a shape trait whose polarity, phylogenetic distribution, and functional significance have been sources of contention. To clarify the determinants of molar flare variation in the hominoid radiation, a combination of statistical methods was employed to investigate the effects of diet, phylogeny, and geologic age upon several measures of molar shape, to identify interactions among these factors, and to estimate their relative influence. Classic indices of molar crown shape and cusp relief are highly significantly associated with diet and show no clear phylogenetic or temporal patterning. Correlations with diet are insignificant when phylogenetic effects are controlled, a result which is interpreted as an artifact of the distribution of folivory in the Miocene hominoid radiation. Possession of pronounced molar flare was found to be the primitive condition for Miocene hominoids, but molar flare reduction cannot be considered a crown hominoid synapomorphy. Molar flare is strongly correlated with geologic age but differs significantly among dietary categories when the effects of time are controlled. Among contemporaneous taxa, hard-object feeders consistently show the highest levels of flare. Molar flare reduction is hypothesized to arise from realignment of cusp positions to maximize molar shearing and increase working occlusal surface area, while variation in flare among contemporaneous taxa may be due, at least in part, to enamel thickness variation. The pronounced molar flare of Otavipithecus is interpreted as a primitive retention, although alternative dietary and phylogenetic interpretations cannot be excluded. A dramatic reversal of molar flare reduction in Mio-Pliocene hominins is interpreted as a synapomorphy of the crown hominin clade, thus supporting the hominin status of the Lukeino hominine. The last common ancestor of the Pan-Homo clade is predicted to have possessed relatively non-flaring molars, and implications of this hypothesis for early hominin recognition are discussed. PMID- 12890446 TI - Enamel microstructure--a truly three-dimensional structure. PMID- 12890447 TI - Aubesier 11 is not evidence of Neanderthal conspecific care. PMID- 12890448 TI - Plasticity of IL-2 and IL-2 receptor chains in rat lymphoid tissues in situ after stimulation with staphylococcal enterotoxin A. AB - Although the effects of mitogens on the synthesis of interleukin-2 (IL-2) and IL 2 receptor (IL-2r) have been described, a detailed in situ analysis of the spatio temporal changes of the expression of the IL-2 gene and the three IL-2r components in lymphoid tissues is still missing. Therefore, we analyzed the IL-2 and IL-2r expression after a staphylococcal enterotoxin A (SEA)-induced T cell activation on a cellular and anatomical basis in the Wistar rat. SEA caused a rapid induction of IL-2 mRNA in T cells of spleen, lymph node, and thymus, followed by the appearance of high systemic IL-2 serum levels (5 ng/ml), and a significant increase of CD25 on CD4+ and CD8+ lymphocytes. The histotopographic analysis of the IL-2r chains revealed a strong upregulation of IL-2r alpha (alpha) and IL-2r beta (beta) mRNAs in similar T cell specific compartments of spleen, lymph node, and thymus as seen for IL-2 mRNA. The abundant constitutive expression of IL-2r gamma (gamma) mRNA was unaffected by SEA. The parallel upregulation of IL-2, IL-2ralpha, and beta chains in conjunction with the continuous presence of the IL-2rgamma chain predominantly in T cell regions of immune organs suggests that the biological effects of IL-2 are essentially limited to T cells, at least after superantigen stimulation. PMID- 12890449 TI - Vascular endothelial growth factor is involved in neoangiogenesis in Hirudo medicinalis (Annelida, Hirudinea). AB - Vascular endothelial growth factor (VEGF) is fundamental in vertebrates for correct development of blood vessels. However, there are only few data about the presence of VEGF in invertebrates. In this study the role of VEGF in neovessel formation is investigated in Hirudo medicinalis. The leech is able to respond to administration of human VEGF by formation of new vessels. The response of H. medicinalis to this growth factor is explained by the presence of two specific VEGF-like receptors (Flt-1/VEGFR-1 and Flk-1/VEGFR-2) as demonstrated by immunohistochemistry and biochemical analysis. The VEGF-like produced by this annelid following surgical stimulation determines not only blood vessel formation, proliferation of vascular endothelial cells but also an increase of cytoplasmic calcium levels. The administration of specific VEGF receptor antibodies can inhibit angiogenesis in leeches previously stimulated with VEGF. PMID- 12890450 TI - Nutritional effects on NF-kappaB/IkappaB activity in weight-gaining and -losing rats. AB - BACKGROUND: We had previously reported metabolic abnormalities in TNF-treated weight-gaining (WG) rats, associated with increased soluble and membrane TNF receptors. OBJECTIVE: On the basis of these findings, we hypothesized that stabilization of TNF in circulation in WG animals by these receptors augmented the action of TNF. TNF influences cellular function through the activation of NF kappaB, by its release from IkappaBalpha and translocation to the nucleus where it regulates gene transcription. This study was designed to determine whether NF kappaB binding was increased in WG rats in comparison with weight-losing (WL) rats. DESIGN: Twenty two male Wistar rats weighing 200-220 g were fed a liquid defined formula diet for 10 days and divided as follows: (i) WG rats (n = 11) receiving an energy density of 0.35 MJ per 60 ml and (ii) WL rats (n = 11) receiving an energy density of 0.09 MJ per 60 ml. Nuclear extracts from heart, liver, lung, spleen and gastrocnemius were used for NF-kappaB measurements by electrophoretic mobility shift assay. Tissue lysates from heart and lung were used for IkappaBalpha measurements by Western blot analysis. RESULTS: There was increased NF-kappaB binding in the lung and heart extracts of the WG rats as compared with WL rats. There was no difference in the binding activity between WG and WL rats in the spleen, liver and gastrocnemius extracts. In lung and heart lysates, IkappaBalpha protein expression was decreased in WG rats as compared with the WL rats. We conclude that there is increased organ-specific TNF activity in WG rats because of increased binding of NF-kappaB to the nucleus and reduced IkappaBalpha levels in the lung and heart. CONCLUSIONS: The findings are consistent with increased TNF activity resulting in the release of NF-kappaB from the NF-kappaB/IkappaBalpha complex and translocation to the nucleus. The nuclear translocation of NF-kappaB observed in the heart and lung suggests that nutrition may influence the action of TNF in these vital organs in sepsis. This finding is potentially of great significance, as pulmonary and cardiac failure are the main causes of death in sepsis. PMID- 12890451 TI - Impact of PGE1 on cyclosporine A induced up-regulation of TGF-beta1, its receptors, and related matrix production in cultured mesangial cells. AB - Transforming growth factor-beta1 (TGF-beta1) plays a major role in cyclosporine A (CsA) induced glomerulosclerosis. We have recently shown that CsA up-regulates the expression of TGF-beta1 and its receptors type I (TbetaR-I) and type II (TbetaR-II) in rat mesangial cells (MCs). Prostaglandins of the E series (PGEs) are known to exert substantial anti-fibrotic effects. Here, we assessed the effect of PGE1 on CsA induced up-regulation of TGF-beta1, TbetaR-I, TbetaR-II and related matrix production in MCs. Co-incubation with PGE1 reduced CsA induced up regulation of TGF-beta1 and TbetaR-II at the mRNA and protein level. Alike, PGE1 reduced TbetaR-I protein expression, which is posttranscriptionally up-regulated by CsA. Whereas a low PGE(1) concentration decreased CsA induced production of fibronectin (FN) and plasminogen activator inhibitor type-1 (PAI-1), a higher PGE1 concentration did not change FN production, but further increased PAI-1 production. In vivo studies will show, whether treatment with PGE1 analogues will be useful in preventing CsA induced glomerulosclerosis. PMID- 12890452 TI - The involvement of IL-6 and IL-8 in acute invasive gastroenteritis of children. AB - The involvement of the proinflammatory cytokines, interleukin 8 (IL-8) and 6 (IL 6), was studied during the first 72 h of acute invasive gastroenteritis. Study population included 33 infants and young children aged six months to six years and seven age-matched controls. As a group, patients with acute invasive gastroenteritis had an increased serum level of IL-8 and IL-6 as compared with healthy controls (p < 0.002 and p < 0.001, respectively). Subjects were then divided into two groups based on stool cultures (proven and non-proven bacterial cultures). Patients with bacterial-proven acute invasive gastroenteritis tended to have increased IL-8 serum concentrations (p < 0.07) as compared with those with non-proven bacterial etiologies and IL-6 levels were only detected in subjects with positive bacterial cultures (p < 0.05). When dividing each sub group into early and late blood drawing with respect to disease onset, no statistical differences were found in each group but subjects with bacterial proven etiologies had significant higher IL-6 levels as compared with non-proven etiologies at the two time points (p < 0.019 and p < 0.015, respectively). In conclusion, the proinflammatory cytokines, IL-6 and IL-8, are involved in acute invasive gastroenteritis. The difference in IL-6, and to a lesser degree IL-8, between proven and non-proven bacterial etiologies, needs further investigation. PMID- 12890453 TI - Serum IL-1beta levels in health and disease: a population-based study. 'The InCHIANTI study'. AB - Interleukin-1 plays a role in normal homeostasis and in the inflammatory response which is deemed to be responsible for the development of major chronic diseases that are highly prevalent in the elderly. Aim of this study is to evaluate the factors influencing the serum levels of Interleukin-1 beta, in a large and representative population. Data were from the InCHIANTI project, a study of factors contributing to the decline of mobility in late life, which sampled people living in two sites in the surroundings of Florence. Blood samples were obtained from 1,292 participants and frozen aliquots were stored at -80 degrees C. The serum levels of several cytokines were measured by enzyme linked immunosorbent assay using an ultrasensitive commercial kit. Interleukin-1 beta serum levels were associated with congestive heart failure (p > 0.001) and angina (p = 0.02), with Ca2+ serum levels (p = 0.02), and with a history of dyslipidemia (p = 0.05). We found no association between serum IL-1beta level and age, sex, consumption of cardioactive drugs and serum levels of IL-1Ra, IL-6, sIL-6R, IL-10 and TNF-alpha. Our data could lend support to the hypothesis that IL-1beta is mainly involved in the functional alterations of cardiomyocytes under conditions marked by mononuclear cell infiltration and by downregulation of calcium. PMID- 12890454 TI - Mechanisms some purchasing authorities in the United Kingdom are using to restrict access to 'cosmetic' surgery within the National Health Service. PMID- 12890455 TI - Clinical decision guidelines for NHS cosmetic surgery: analysis of current limitations and recommendations for future development. AB - Because of increasing demand for publicly funded elective cosmetic surgery, clinical decision guidelines have been developed to select those patients who should receive it. The aims of this study were to identify: the main characteristics of such guidelines; whether and how they influence clinical decision making; and ways in which they should be improved. UK health authorities were asked for their current guidelines for elective cosmetic surgery and, in a single plastic surgery unit, we examined the impact of its guidelines by observing consultations and interviewing surgeons and managers. Of 115 authorities approached, 32 reported using guidelines and provided sufficient information for analysis. Guidelines mostly concerned arbitrary sets of cosmetic procedures and lacked reference to an evidence base. They allowed surgery for specified anatomical, functional or symptomatic reasons, but these indications varied between guidelines. Most guidelines also permitted surgery 'exceptionally' for psychological reasons. The guidelines that were studied in detail did not appreciably influence surgeons' decisions, which reflected criteria that were not cited in the guidelines, including cost of the procedure and whether patients sought restoration or improvement of their appearance. Decision guidelines in this area have several limitations. Future guidelines should: include all cosmetic procedures; be informed by a broad range of evidence; and, arguably, include several nonclinical criteria that currently inform surgeons' decision making. PMID- 12890456 TI - Perception and reality-a study of public and professional perceptions of plastic surgery. AB - A questionnaire survey of the perception of plastic surgery amongst 1567 members of the public, general practitioners and medical students is presented. Closed ended format questions were designed to assess understanding of the range of conditions managed by plastic surgeons. Respondents were asked to match nine surgical specialists with 40 conditions or procedures. To investigate understanding of the multidisciplinary nature of some surgery, respondents were asked which type of surgeon might have a supplementary role. Completed questionnaires from 1004 members of the public, 335 general practitioners, and 228 medical students are presented (responses rate>65%). Significant differences were identified between public respondents and other groups. Plastic surgery was associated with reconstruction for trauma and cancer and procedures with a strong aesthetic element by all three groups. The public were poorly informed about some core plastic surgery including burns, melanoma and hand surgery. General practitioner and student respondents had a better understanding of the diversity of the specialty. However, both groups considered orthopaedic surgeons and not plastic surgeons to be hand surgeons. The strengths and weaknesses of this study are discussed together with potential areas for education and promotion. PMID- 12890457 TI - Exogenous leukaemia inhibitory factor enhances nerve regeneration after late secondary repair using a bioartificial nerve conduit. AB - The clinical outcome of peripheral nerve injuries remains disappointing, even in the ideal situation of a primary repair performed with optimal microsurgical techniques. Primary repair is appropriate for only about 85% of injuries, and outcome is worse following secondary nerve repair, partly owing to the reduced regenerative potential of chronically axotomised neurons. Leukaemia inhibitory factor (LIF) is a gp-130 neurocytokine that is thought to act as an 'injury factor', triggering the early-injury phenotype within neurons and potentially boosting their regenerative potential after secondary nerve repair. At 2-4 months after sciatic nerve axotomy in the rat, 1 cm gaps were repaired using either nerve isografts or poly-3-hydroxybutyrate conduits containing a calcium alginate and fibronectin hydrogel. Regeneration was determined by quantitative immunohistochemistry 6 weeks after repair, and the effect of incorporating recombinant LIF (100 ng/ml) into the conduits was assessed. LIF increased the regeneration distance in repairs performed after both 2 months (69%, P=0.019) and 4 months (123%, P=0.021), and was statistically comparable to nerve graft. The total area of axonal immunostaining increased by 21% (P>0.05) and 63% (P>0.05), respectively. Percentage immunostaining area was not increased in the 2 months group, but increased by 93% in the repairs performed 4 months after axotomy. Exogenous LIF, therefore, has a potential role in promoting peripheral nerve regeneration after secondary repair, and can be effectively delivered within poly 3-hydroxybutyrate bioartificial conduits used for nerve repair. PMID- 12890458 TI - Hidradenitis suppurativa: pathogenesis and management. AB - Hidradenitis suppurativa (HS) is a chronic disease manifested by recurrent abscesses, sinus tracts and scarring. It arises most commonly, however, not exclusive from apocrine gland bearing skin. This review article sets out to clarify the importance of early diagnosis, the pathogenesis and aetiology of HS, and evidence for medical and surgical therapies for this debilitating disease.HS is caused primarily by follicular occlusion with secondary involvement of the apocrine glands. The aetiology is still poorly understood. There is a genetic component with probable hormonal influence on gene expression. Shearing forces from obesity and tight clothing contribute to its development. Management should be appropriately tailored for the severity and distribution of HS as well as quality of life of the patient. Medical management with appropriate antibiotics, if initiated early, can be successful in mild to moderate severity HS as well as improving disease control prior to attempted curative surgery in severe HS. Other helpful measures include advice on lifestyle changes, intralesional steroids, systemic retinoids, hormonal manipulation, and a revival of interest in the use of radiotherapy for HS. While there is a place for 'conservative' surgical procedures (including CO(2) laser) in selected cases of mild to moderate HS, radical excision of all apocrine-bearing tissue is the definitive treatment. We advocate close interdisciplinary collaboration as well as a cautionary approach to timing and planning of surgery to minimise recurrence rates. PMID- 12890459 TI - Better cosmetic results and comparable quality of life after skin-sparing mastectomy and immediate autologous breast reconstruction compared to breast conservative treatment. AB - Preoperative chemotherapy (PCT) can be used in large primary breast cancer to facilitate breast conservative surgery (BCS). Cosmetic results of BCS are influenced by the size of the residual tumour, relative to the size of the breast. After mastectomy, immediate breast reconstruction (IBR) with autologous tissue provides excellent cosmetic outcome and has proven to be safe in breast cancer patients. Besides improving overall and disease free survival, Quality of Life (QoL), body image and cosmetic outcome are also important issues after treatment for breast cancer. In this study, Health-Related-Quality of Life (HRQL) and body image were evaluated, in patients treated with PCT, followed by BCS, or skin-sparing mastectomy (SSM) and perforator-flap breast reconstruction. Additionally, clinical observers assessed cosmetic outcome. All participants were evaluated by the Medical Outcomes Study (MOS) 36-item Short Form Health Status Survey (SF-36, 36 items) and a study-specific questionnaire. An external panel evaluated standardised photographs of the breasts. For all patients, norm-based scores of physical and mental health state are comparable with the general population, except for vitality (VT) score, which is somewhat lower. No significant differences can be observed between both groups. The majority of the patients were satisfied with the appearance of their breasts. The cosmetic results, assessed by the clinical team, were significantly better for patients having IBR, compared to BCS. The mean score was 7.5/10 for IBR, versus 6.0/10 for BCS (p<0.0001).Breast conserving treatment or mastectomy with reconstruction may yield comparable results of QoL, but cosmetic outcome is better after SSM and perforator-flap reconstruction. Patients must be offered both options, and clinicians should stress that both are equally effective. PMID- 12890460 TI - The pedicled occipital artery scalp flap for salvage surgery of the neck. AB - A small group of patients with complex head and neck cancer present with problems of wound healing following radiotherapy and reconstructive surgery. Provision of skin cover to the neck in these cases is often required and presents a challenge to the reconstructive surgeon. We present the use of a pedicled scalp flap based on the occipital artery for such defects. This flap is an axial patterned scalp flap incorporating hair-bearing skin. It may be up to 15 cm wide and can reach beyond the midline of the chin. The anatomy of the flap is described and its use illustrated in three cases. This flap is a useful addition to the options for reconstruction of neck defects in patients with head and neck cancer. PMID- 12890461 TI - Concept for postoperative analgesia after pedicled TRAM flaps: continuous wound instillation with 0.2% ropivacaine via multilumen catheters. A report of two cases. AB - Pedicled TRAM flap surgery is a complex procedure characterised by an extensive wound site. We present two patients with efficient postoperative pain relief by continuous wound instillation of ropivacaine 0.2% via two multilumen catheters. The catheters were placed subcutaneously before the wound closure through the umbilicus into the abdominal wound, and under the autologous flap into the breast. Each multilumen catheter provides even distribution for local anaesthetics over 12.5 cm. At the end of surgery, patients received a single shot dose of local anaesthetic via the pain catheters. After surgery the continuous infusion of ropivacaine 0.2% was commenced at a rate of 10 ml/h per catheter. Pain scores at rest and on coughing were low on the first postoperative day, and later zero. No medication for breakthrough pain was required throughout the recovery period, and the patients experienced no adverse events linked to the analgesia scene. Patient satisfaction was excellent, and quality of recovery score was superior. PMID- 12890462 TI - Spermatogenesis after scrotal reconstruction. AB - Objective. To explore the effect of scrotal reconstruction following avulsion on spermatogenesis. Methods. Three patients were followed-up after having their scrotums reconstructed. Two scrotums were reconstructed by skin flap. Another patient's testicles were buried in thigh pockets. The follow-up continued from 3 months to 4 years after operation, and we examined the sperm quality, sexual function, sexual hormone, and testis biopsy. Results. The follow-up data showed that the spermatogenesis was not altered in the early stage (up to three months), but was substantially abnormal in the late stage (after two years). Conclusions. Using the thick skin flaps, or burying the testicle for scrotum reconstruction are not recommended for patients who wish to remain fertile. PMID- 12890463 TI - Single-stage muscle flap reconstruction of major scrotal defects: report of two cases. AB - Major scrotal defect with exposed testes and/or spermatic cords are a challenge for the reconstructive surgeon. The bacterial flora of the perineum, difficulty of immobilisation and the contour of the testes make testicular cover a difficult task [Br. J. Plast. Surg. 41 (1988) 190]. Traditional approaches have used simple skin grafts or if not feasible, multi-staged procedures with initial burying of the testes under delayed medial thigh flaps. Better techniques then evolved to permit early single-staged coverage using flaps rather than skin grafts in these usually contaminated and unsuitable wounds to improve the cosmetic outcome of the reconstruction and reduce patient discomfort and hospital stay. Muscle flaps represent an excellent reconstruction option in the contaminated perineum especially in patients with impaired ability to deal with infection such as diabetic or toxic patients. We present two cases of reconstruction of the scrotum using simple, reliable single-stage muscle flap techniques with good aesthetic results and review the literature. PMID- 12890464 TI - Tie-overs under pressure. AB - Dressings tied over skin grafts reduce the size of dead space, prevent haematoma formation, and immobilize the grafts [McCarthy, Plastic Surgery, 1990]. Their mechanism of action was thought to include the application of pressure to the bed and stabilization of the graft and dressing. A quantitative analysis of the pressure exerted by these dressings has never been reported [Wolf at al., Ann Plast Surg 40 (1998) 149]. This study measures the pressure exerted by tie-over dressings at the graft-bed interface and finds that no significant pressure is exerted. PMID- 12890465 TI - Adhesive retention dressings are more comfortable than alginate dressings on split-skin-graft donor sites. AB - Painful split-skin-graft donor sites remain a common problem for patients. We undertook a prospective randomised trial to examine the comparative comfort and ease of care of two different donor-site dressings. One dressing is the alginate Kaltostat, the standard plastic-surgical dressing in the UK and abroad, and the other is the adhesive retention tape Mefix, a novel use of a readily available dressing. We randomised 50 patients requiring split-skin grafts to receive either alginate (Kaltostat) or retention (Mefix) donor-site dressings. Dressings were assessed by interview and questionnaire at 24, 72 h and 2 weeks, and by wound review at 2 weeks. Retention dressings were found to be more comfortable, required less nursing intervention and allowed patients easier mobility with a greater range of daily activities, especially washing, without compromising wound healing. We recommend adhesive retention dressings as cost-effective comfortable dressings, which readily conform to any donor site. PMID- 12890466 TI - Replantation of multi-level fingertip amputation using the pocket principle (palmar pocket method). AB - Two cases of multi-level fingertip amputation are presented. In each case, replantation was achieved in a two-stage procedure, involving reattachment, de epithelialisation and insertion into a palmar pocket in stage 1, followed by removal from the palmar pocket 16 days later. The cases are described and the technique is discussed. PMID- 12890467 TI - Thumb reconstruction for aesthetic reasons using an exteriorised microvascular pedicle. AB - Thumb reconstruction following amputation is usually performed in order to restore function. Nevertheless, the reconstruction should be cosmetically acceptable in order to be fully functional, and, in some cases, reconstructive surgery may be justified for purely aesthetic reasons. The most satisfying aesthetic results in adults are obtained with microsurgical partial great-toe transfer. The technique that we use for thumb reconstruction is illustrated by two case reports: that of a 26-year-old female patient and that of a 35-year-old male patient. Both patients had a distal thumb amputation with destroyed nail apparatus, and both sought thumb reconstruction for cosmetic reasons. Aesthetic reconstruction was performed in both cases with a partial ipsilateral great-toe transfer, composed of nail apparatus, underlying bone and custom-made pulp tissue. The vascular anastomosis was done at the snuff-box, through a small incision, with an exteriorised pedicle. The result was satisfactory in both cases, with minimal donor-site sequelae. Partial toe transfer has proven to be a reliable technique for thumb reconstruction. It is an evolving technique. Many modifications have been introduced to optimise the aesthetic result and to reduce donor-site morbidity. Our technique allows us to restore thumb length, replace the missing nail and reconstruct the pulp, with acceptable sequelae at the donor toe. The exteriorised-pedicle technique prevents pedicle compression and twisting and reduces scarring and stiffness. It does, however, require delicate postoperative care and a second procedure for pedicle division. PMID- 12890468 TI - Massive plexiform neurofibroma with associated meningo-encephalocoele and occipital bone defect presenting as a cervical mass. AB - The commonest skull manifestations in neurofibromatosis involve the orbit, with very few reports about occipital defects. We report a rare case of a 54-year-old lady with a massive plexiform neurofibroma extending from the auricular region down her left neck and into her shoulder, with an associated large left occipital and left petrous bone defect and extensive cerebellar meningo-encephalocoele, which presented with a relatively asymptomatic cervical mass and was treated with resection of the neurofibroma and advancement and rotational skin flaps. PMID- 12890469 TI - Leakage and silicone lymphadenopathy with cohesive breast implant. PMID- 12890470 TI - Histological appearances of Trilucent breast implant capsules. PMID- 12890471 TI - Composition and function of the eukaryotic N-terminal acetyltransferase subunits. AB - Saccharomyces cerevisiae contains three N-terminal acetyltransferases (NATs), NatA, NatB, and NatC, composed of the following catalytic and auxiliary subunits: Ard1p and Nat1p (NatA); Nat3p and Mdm20p (NatB); and Mak3p, Mak10, and Mak31p (NatC). The overall patterns of N-terminally acetylated proteins and NAT orthologous genes suggest that yeast and higher eukaryotes have similar systems for N-terminal acetylation. The differential expression of certain NAT subunits during development or in carcinomas of higher eukaryotes suggests that the NATs are more highly expressed in cells undergoing rapid protein synthesis. Although Mak3p is functionally the same in yeast and plants, findings with TE2 (a human Ard1p ortholog) and Tbdn100 (a mouse Nat1p ortholog) suggest that certain of the NAT subunits may have functions other than their role in NATs or that these orthologs are not functionally equivalent. Thus, the vertebrate NATs remain to be definitively identified, and, furthermore, it remains to be seen if any of the yeast NATs contribute to other functions. PMID- 12890472 TI - alpha 2B-Adrenoceptor levels govern agonist and inverse agonist responses in PC12 cells. AB - Receptor density is an important determinant of cellular effector responses to receptor activation. We analysed cytosolic Ca(2+) responses to alpha(2) adrenergic agents in PC12 cells expressing human alpha(2B)-adrenergic receptors (AR) at two densities (3.8 and 1.3 pmol/mg protein). The efficacy (E(max)) of agonists was greater in cells with higher receptor expression; while the potency (EC(50)) of norepinephrine and oxymetazoline was independent of alpha(2B)-AR levels. Several classical alpha(2)-AR antagonists behaved as either partial or inverse agonists in a receptor density-dependent fashion. No apparent structural similarities were found among the inverse agonists, precluding simple predictions of inverse agonist activity. Transfected PC12 cells expressing alpha(2B)-AR at relatively high density would be a useful approach to screen inverse agonists for this class of receptors. Our results further indicate that receptor density significantly influences the properties of ligands, not only of partial agonists as predicted by classical receptor theory, but also of antagonists and full agonists. PMID- 12890473 TI - A peptide mimetic of Gal-alpha 1,3-Gal is able to block human natural antibodies. AB - The carbohydrate of Gal-alpha1,3-Gal is thought to be the major antigenic epitope present on pig vascular endothelium. The peptides that mimic the binding of antigenic epitope (Gal-alpha1,3-Gal) to lectin BS-I-B4 were identified from screening a filamentous phage-displayed random library. A phage bearing the peptide NCVSPYWCEPLAPSARA has been identified to bind the lectin strongly. Melibiose was able to inhibit the binding of the human natural anti-alpha Gal antibody to the peptide competitively. Our experiments show that the peptide mimetic of Gal-alpha1,3-Gal is able to inhibit the agglutination of pig RBCs by human natural antibody or lectin BS-I-B4. The peptide inhibitor of human natural antibodies may prove useful in pig-to-human xenotransplantation. PMID- 12890474 TI - Neuregulin-1 suppresses muscarinic receptor expression and acetylcholine activated muscarinic K+ channels in cardiac myocytes. AB - The neuregulin-1 family of growth factors regulates nicotinic acetylcholine receptor synthesis in skeletal muscle, but its role in cardiac myogenesis remains unclear. Here, we investigate the involvement of neuregulins in the development of cardiac cholinergic responsiveness. Treatment of chick cardiac myocytes with neuregulin-1 inhibited mRNA expression of the M4 muscarinic receptor, but not the M2 receptor. In addition, mRNA levels of GIRK1 were reduced in myocytes by treatment with neuregulin-1. Activation of cholinergic receptors in cultured chick atrial myocytes by carbachol produced an outward potassium current (I(K(ACh))), which was attenuated by 24-48-h pre-treatment with neuregulin-1. These data suggest that neuregulins can regulate cardiac parasympathetic tone and may be involved in the pathogenesis of cardiac arrhythmias and heart failure. PMID- 12890475 TI - Calorie restriction in mice does not affect LDL reverse cholesterol transport in vivo. AB - Calorie restriction (CR) prolongs life in animals, but may reduce plasma HDL, important in reverse cholesterol transport (RCT). The effect of CR, 60% of an ad libitum (AL) diet, on cholesterol removal from rectus femoris muscle injected with cationized LDL, was studied in C57BL male mice. RCT in vivo, on CR and AL diet, and cholesterol efflux from macrophages exposed to CR or AL sera, was similar, despite a 22% reduction in plasma HDL-cholesterol (HDL-C). In CR fed mice total cholesterol (TC) and phospholipid (T-PL) decreased by 32% and 38%, while HDL-C and HDL-PL decreased by 22% and 16% only, resulting in increased HDL PL/T-PL ratio, which enhanced RCT. Partial re-feeding (CR-RF, 70% of AL) induced normalization of plasma lipids (excluding triglycerides), while HDL-PL/T-PL remained elevated. Thus, as CR did not interfere with RCT in vivo, it could possibly be beneficial to patients at risk for coronary heart disease. PMID- 12890476 TI - Aluminum-induced distortion in calcium signaling involving oxidative bursts and channel regulation in tobacco BY-2 cells. AB - Trivalent cations such as those of Al, La, and Gd are phytotoxic. Our previous works showed that addition of LaCl(3) or GdCl(3) to tobacco cells triggers the generation of superoxide (O(2)*-). Here, we show that AlCl(3) at normal physiological pH (5.8) induces much greater production of O(2)*- (detected with a specific chemiluminescence probe), indicating that these trivalent cations similarly induce the oxidative bursts. It was shown that NADPH oxidase is involved in the generation of O(2)*- and the yield of O(2)*- was dose-dependent (ca. 6mM Al, optimal). Following the acute spike of O(2)*-, a gradual increase in cytosolic-free Ca(2+) concentration ([Ca(2+)](c)) was detected with the luminescence of recombinant aequorin over-expressed in the cytosol. Interestingly, a O(2)*- scavenger and a Ca(2+) chelator significantly lowered the level of [Ca(2+)](c) increase, indicating that the Al-induced O(2)*- stimulates the influx of Ca(2+). Compared to the induction of O(2)*- generation, the [Ca(2+)](c) elevation was shown to be maximal (340 nM) at relatively lower Al concentrations (ca. 1.25 mM). Thus, the Al concentration optimal for O(2)*- is too much (inhibitory) for [Ca(2+)](c). In addition, high concentrations of Al were shown to be inhibitory to the H(2)O(2)-induced Ca(2+) influx. This explains the ineffectiveness of high Al concentration in the oxidative burst-mediated induction of [Ca(2+)](c) increase. It is likely that Al-induced [Ca(2+)](c) elevation is manifested from the finely geared balance between the O(2)*- mediated driving force and the channel inhibition-mediated brake. Furthermore, it is note-worthy that Al (< or =10mM) showed no inhibitory effect on the hypo osmolarity-induced Ca(2+) influx, implying that Al may be a selective inhibitor of redox-responsive Ca(2+) channels. Possible target channels of Al actions are discussed. PMID- 12890477 TI - Expression of Class III facilitative glucose transporter genes (GLUT-10 and GLUT 12) in mouse and human adipose tissues. AB - We have examined whether GLUT-10 and GLUT-12, members of the Class III group of the recently expanded family of facilitative glucose transporters, are expressed in adipose tissues. The mouse GLUT-12 gene, located on chromosome 10, comprises at least five exons and encodes a 622 amino acid protein exhibiting 83% sequence identity and 91% sequence similarity to human GLUT-12. Expression of the GLUT-12 gene was evident in all the major mouse adipose tissue depots (epididymal, perirenal, mesenteric, omental, and subcutaneous white; interscapular brown). The GLUT-10 gene is also expressed in mouse adipose tissues and as with GLUT-12 expression occurred in the mature adipocytes as well as the stromal vascular cells. 3T3-L1 adipocytes express GLUT-10, but not GLUT-12, and expression of GLUT 12 was not induced by insulin or glucose. Both GLUT-10 and GLUT-12 expression was also found in human adipose tissue (subcutaneous and omental) and SGBS adipocytes. It is concluded that white fat expresses a wide range of facilitative glucose transporters. PMID- 12890478 TI - Small molecule inhibitors of the RNA-dependent protein kinase. AB - The RNA-dependent protein kinase (PKR) is an interferon-induced serine/threonine protein kinase that phosphorylates the alpha subunit of the eukaryotic initiation factor 2 in response to viral infection. Classical genetic approaches for studying the role of PKR in cell signaling have their limitations due to overlapping but non-redundant pathways. Small molecule inhibitors of PKR will be useful in this regard. We report here, the discovery of a small molecule inhibitor of the kinase reaction of PKR. The inhibitor was discovered by screening a library of 26 different ATP-binding site directed inhibitors of varying structure. We also describe the development of a high-throughput assay for screening a large number of compounds for a PKR inhibitor using a rabbit reticulocyte lysate system and luciferase mRNA. The assay takes advantage of the fact that the reticulocyte lysate is rich in components of the translational machinery, of which PKR is an integral part. This assay can be carried out with added exogenous human PKR to study the effect of various compounds in their ability to rescue the translational block imposed by human PKR. PMID- 12890479 TI - Development of transgenic fish for ornamental and bioreactor by strong expression of fluorescent proteins in the skeletal muscle. AB - In the present study, new applications of the transgenic technology in developing novel varieties of ornamental fish and bioreactor fish were explored in a model fish, the zebrafish (Danio rerio). Three "living color" fluorescent proteins, green fluorescent protein (GFP), yellow fluorescent protein (YFP), and red fluorescent protein (RFP or dsRed), were expressed under a strong muscle-specific mylz2 promoter in stable lines of transgenic zebrafish. These transgenic zebrafish display vivid fluorescent colors (green, red, yellow, or orange) visible to unaided eyes under both daylight and ultraviolet light in the dark. The level of foreign protein expression is estimated between 3% and 17% of total muscle proteins, equivalent to 4.8-27.2mg/g wet muscle tissue. Thus, the fish muscle may be explored as another useful bioreactor system for production of recombinant proteins. In spite of the high level of foreign protein expression, the expression of endogenous mylz2 mRNAs was not negatively affected. Furthermore, compared to the wild-type fish, these fluorescent transgenic fish have no advantage in survival and reproduction. PMID- 12890480 TI - Effect of green tea polyphenols on angiogenesis induced by an angiogenin-like protein. AB - Angiogenesis is a fundamental process by which new blood vessels are formed. The angiogenesis process is induced by several growth factors. Among them angiogenin is the most potent blood vessel inducer known. In this paper, we have investigated the effect of green tea polyphenols, mainly the catechins, on an angiogenin-like protein induced angiogenesis process. The angiogenin-like protein was isolated from goat serum and the effect of green tea components was tested by the chicken chorioallantoic membrane (CAM) assay. The results show that green tea components are capable of reducing the vascularization on CAM that is induced by the angiogenin-like protein. PMID- 12890481 TI - Structural determinant for cold inactivation of rodent L-xylulose reductase. AB - L-Xylulose reductase (XR) is a homotetramer belonging to the short-chain dehydrogenase/reductase family. Human XR is stable at low temperature, whereas the enzymes of mouse, rat, guinea pig, and hamster are rapidly dissociated into their inactive dimeric forms. In order to identify amino acid residues that cause cold inactivation of the rodent XRs, we have here selected Asp238, Leu242, and Thr244 in the C-terminal regions of rodent XRs and performed site-directed mutagenesis of the residues of mouse XR to the corresponding residues (Glu, Trp, and Cys) of the human enzyme. Cold inactivation was prevented partially by the single mutation of L242W and the double mutation of L242W/T244C, and completely by the double mutation of D238E/L242W. The L242W and L242W/T244C mutants existed in both tetrameric and dimeric forms at low temperature and the D238E/L242W mutant retained its tetrameric structure. No preventive effect was exerted by the mutations of D238E and T244C, which were dissociated into their dimeric forms upon cooling. Crystallographic analysis of human XR revealed that Glu238 and Trp242 contribute to proper orientation of the guanidino group of Arg203 of the same subunit to the C-terminal carboxylate group of Cys244 of another subunit through the neighboring residues, Gln137 and Phe241. Thus, the determinants for cold inactivation of rodent XRs are Asp238 and Leu242 with small side chains, which weaken the salt bridges between Arg203 and the C-terminal carboxylate group, and lead to cold inactivation. PMID- 12890482 TI - Kinetic behavior of Desulfovibrio gigas aldehyde oxidoreductase encapsulated in reverse micelles. AB - We report the kinetic behavior of the enzyme aldehyde oxidoreductase (AOR) from the sulfate reducing bacterium Desulfovibrio gigas (Dg) encapsulated in reverse micelles of sodium bis-(2-ethylhexyl) sulfosuccinate in isooctane using benzaldehyde, octaldehyde, and decylaldehyde as substrates. Dg AOR is a 200-kDa homodimeric protein that catalyzes the conversion of aldehydes to carboxylic acids. Ultrasedimentation analysis of Dg AOR-containing micelles showed the presence of 100-kDa molecular weight species, confirming that the Dg AOR subunits can be dissociated. UV-visible spectra of encapsulated Dg AOR are indistinguishable from the enzyme spectrum in solution, suggesting that both protein fold and metal cofactor are kept intact upon encapsulation. The catalytic constant (k(cat)) profile as a function of the micelle size W(0) (W(0)=[H(2)O]/[AOT]) using benzaldehyde as substrate showed two bell-shaped activity peaks at W(0)=20 and 26. Furthermore, enzymatic activity for octaldehyde and decylaldehyde was detected only in reverse micelles. Like for the benzaldehyde kinetics, two peaks with both similar k(cat) values and W(0) positions were obtained. EPR studies using spin-labeled reverse micelles indicated that octaldehyde and benzaldehyde are intercalated in the micelle membrane. This suggests that, though Dg AOR is found in the cytoplasm of bacterial cells, the enzyme may catalyze the reaction of substrates incorporated into a cell membrane. PMID- 12890483 TI - Active repression of organizer genes by C-terminal domain of PV.1. AB - PV.1, a homeotic protein, ventralizes dorsal mesoderm and inhibits neuralization by mediating BMP-4 signaling in Xenopus embryo. In our previous report antimorphic PV.1 causes a secondary axis by inducing the ectopic organizer. We analyzed the structure of this transcription factor through domain level assessment. In a phenotype-inducing test, half of the N-terminus at the N terminal side was unessential for inducing ventralization of embryos. We examined the transacting activity of several regions of PV.1 utilizing GAL4 hybrid system. The C-terminal region/GAL4DBD (DNA binding domain) exhibited strong repressive activity on a reporter gene (operator/promoter/reporter; Gal4-TK-luc) as much as the whole polypeptide/GAL4DBD, whereas the N-terminal region/GAL4DBD showed only modest repression. The results suggest that PV.1 functions as a transcriptional repressor and this repressive activity is localized mostly to the C-terminal region. Additional characterizations of N- and C-terminus with respect to the effects on the expression of other genes are described. PMID- 12890484 TI - Function and structure of recombinant single chain calcineurin. AB - Calcineurin (CN) is a Ca(2+)/calmodulin(CaM)-dependent serine/threonine protein phosphatase which is a heterodimer composed of a 61 kDa catalytic subunit (CNA) and a 19 kDa regulatory subunit (CNB). The enzyme is critical for several important intracellular signal-transducing pathways, including T-cell activation. Its crystal structure reveals that the C-terminal of CNB lies in close vicinity of the N-terminal of CNA and each end has a long arm not involved in the active site. After fusing two subunits, it was determined that folding and function of the protein were not affected by the fusion. We amplified a fused gene of A and B subunits using a pair of linker primers including six codons of glycine. A single chain calcineurin was constructed and purified to near-homogeneity. The recombinant enzyme was fully soluble, displayed high specific activity with substrate, and exhibited biochemical properties and kinetic parameters similar to those of the native enzyme from the bovine brain. It was still activated by Ca(2+)/calmodulin but was not regulated by extra CNB and was still strongly stimulated by Mn(2+) and Ni(2+) divalent metal ions. The solution conformations of both recombinant enzyme and bovine calcineurin were assayed under the same conditions using intrinsic fluorescence spectroscopy and circular dichroism spectropolarimetry, and results showed their graphs are approximately identical. Our findings suggested that the fusion of A and B subunits of calcineurin does not affect their folding pathways and structural changes involved in their function, furthermore, they are bound to the correct binding site. PMID- 12890485 TI - Large-scale purification of an antibody directed against hepatitis B surface antigen from transgenic tobacco plants. AB - The application of bioengineering to plants for production of biological products for human and animal use has expanded in recent years. The reasons for this expansion are several and include advances in the technology for novel production systems and the need for very large quantities of therapeutic proteins. The process of growing pharmaceutical proteins in plants, extracting, and purifying is a hard task considering the lack of available information concerning these topics. In this work, a recombinant murine monoclonal antibody specific for the hepatitis B surface antigen, expressed in stably transformed transgenic Nicotiana tabacum plants, was purified by means of a recombinant protein A Streamline chromatography as the main purification step. The antibody expression level varied with the age of the plants and the number of harvests from 40 to 15microg/ml and the maximum process yield was about 25mg of plantibody/kg of biomass. Protein A Streamline chromatography was successfully used in the purification process yielding a recovery of about 60% and a plantibody SDS-PAGE purity of over 90% but unexpectedly, previous clarification steps could not be totally avoided. The amino acid sequence recognized by this affinity purified plantibody was similar to its murine counterpart verifying the potentiality of plants to replace animals or bioreactors for large-scale production of this monoclonal antibody. PMID- 12890486 TI - Localization of Tie2 and phospholipase D in endothelial caveolae is involved in angiopoietin-1-induced MEK/ERK phosphorylation and migration in endothelial cells. AB - Angiopoietin-1 (Ang1) and its receptor, Tie2, play critical roles in blood vessel formation. Ang1 triggers a variety of signaling events in endothelial cells leading to vasculogenic and angiogenic processes. However, the underlying mechanism for Ang1/Tie2 signaling is not fully understood. Here, we show that Tie2 and phospholipase D (PLD) are localized in the caveolae, specialized subdomains of the endothelial cell plasma membrane enriched with signaling molecules. Interestingly, Ang1 increased PLD activities in a dose- and time dependent manner. Ang1-induced MEK/ERK activation was abrogated when PLD was inhibited, suggesting that PLD mediates Ang1-induced MEK/ERK activation. Moreover, PLD inhibitor, 1-butanol, inhibited Ang1-induced endothelial cell migration. Our results indicate that: (1) caveolae may be the platform for Tie2/PLD association in endothelial cells; (2) PLD is a new mediator of Ang1/Tie2 induced signaling pathway, and it participates in MAPK activation and endothelial cell migration. PMID- 12890487 TI - A novel HECT-type E3 ubiquitin ligase, NEDL2, stabilizes p73 and enhances its transcriptional activity. AB - Expression of p73, a p53 family member regulating cell growth and apoptosis, is maintained at low levels in mammalian cells, and cellular activation of p73 is usually controlled at the protein level. However, the precise molecular mechanisms by which p73 stability is regulated are unclear. During the search for interacting molecules with the COOH-terminal proline-rich region of p73, we identified a novel NEDD4-related protein (termed as NEDL2) which contains a C2 domain at its NH(2)-terminus, two WW domains, and a HECT domain at its COOH terminus. As expected, NEDL2 catalyzed the ubiquitination of bacterial cellular proteins in vitro. Reciprocal co-immunoprecipitation experiments and in vitro pull-down assays revealed that NEDL2 bound to p73, which carries two putative PY motifs. p73 was efficiently ubiquitinated but stabilized in a NEDL2-dependent manner. Accordingly, p73 decayed at faster rates in the absence of NEDL2 than in its presence. Consistent with the NEDL2-mediated stabilization of p73, NEDL2 enhanced the p73-dependent transcriptional activation. Thus, our results suggest that NEDL2 activates the function of p73 by increasing its stability. PMID- 12890488 TI - The endogenous inhibitor of NCX1 does not resemble the properties of digitalis compound. AB - In our previous study, we ware successful in isolation and purification of an endogenous inhibitor of the Na/Ca exchanger (NCX1) from the calf ventricle extracts. The purified factor has characterized to have strong positive inotropic effect on isometric contractions of isolated ventricle strips of guinea pig. A possibility is that besides the NCX1 the endogenous factor may also interact with other ion-transport systems (e.g., Na,K-ATPase) involved in modulation of muscle contractility-relaxation. Therefore, a primary goal of the present study was to detect a possible effect of newly found NCX1 inhibitor on Na,K-ATPase and Ca ATPase activities. The preparations of isolated sarcolemma vesicles were used for this goal. Although the crude extracts of calf ventricles can inhibit both the Na/Ca exchange and Na,K-ATPase, these two inhibitory activities can be separated on the Sephadex G-10 column, meaning that different molecular entities might be responsible for inhibition of Na/Ca exchange and Na,K-ATPase. Addition of 100 U of purified endogenous factor to the assay medium results in nearly complete inhibition of forward (Na(i)-dependent Ca-uptake) and reverse (Na(o)-dependent Ca efflux) modes of Na/Ca exchange. On the other hand, no effect was detected on activities of Na,K-ATPase and Ca-ATPase even in the presence of 500 U of purified factor in the assay medium. In light of the present data, it is concluded that the endogenous inhibitor of NCX1 does not resemble the targeting properties of digitalis like compound. Obviously, more systematic studies are required in the future for resolving a possible interaction of the endogenous inhibitor of NCX1 with other ion-transport systems involved in calcium homeostasis and action potential. PMID- 12890489 TI - PI3K inhibitors changed the p53-induced response of Saos-2 cells from growth arrest to apoptosis. AB - p53 is activated by stress leading to oncogenic alteration, which induces either cell cycle arrest or apoptosis, although the mechanism involved in this decision has not been fully clarified as yet. This work was undertaken to change the cellular response by inducing apoptosis with PI3K inhibitors to Saos-2 cells that had been growth-arrested in both G1 and G2/M by the wild-type activity of temperature-sensitive (ts) p53. We found that the PI3K/Akt inhibitors LY294002 and wortmannin, but not the MEK inhibitor U0126, were capable of inducing apoptosis in growth-arrested Saos-2 cells, as assessed by an increase in the sub G1 population, pyknotic nuclei, and DNA ladder formation. We detected the cleavage of caspases 9 and 3, and PARP after LY294002 addition, accompanied by a loss of cytochrome c from the mitochondria, and observed Bax translocation to the mitochondria and down-regulation of phospho-Akt, suggesting that blocking of survival signals triggered the apoptotic signal through the mitochondrial apoptotic pathway. It is thus suggested that the PI3K/Akt pathway played an important role in determining cell fate between growth arrest and apoptosis. PMID- 12890490 TI - A novel murine PDGF-D splicing variant results in significant differences in peptide expression and function. AB - Platelet-derived growth factor (PDGF) is a potent mesenchymal cell mitogen and chemoattractant involved in the pathogenesis of fibroproliferative diseases. There are four known PDGF ligand isoforms designated A-D, two of which, C and D, were only recently discovered. We have identified a splicing variant in the PDGF D isoform that occurs in mice, but not in humans. The presence of the splicing variant in murine PDGF-D appears to be due to an aberration in the splicing site at the junction of exons 5 and 6. The splicing variant results in a deletion predicted to have significant effects on peptide activity since it results in the deletion of bases within the cysteine knot domain that are important for peptide dimerization and receptor binding. It is important to appreciate differences between murine and human PDGF gene expression because PDGF is a key mitogen in the pathogenesis of fibrosis and mice are commonly employed as models for human disease. PMID- 12890491 TI - PCR-based subtraction analyses for upregulated gene transcription in cadmium exposed rat lung type 2 epithelial cells. AB - The aim of this study was to gain insight into early events in the lung epithelial cells following acute Cd exposure. We adopted the polymerase chain reaction (PCR)-based subtraction technique and found several genes that were upregulated in immortalized rat lung type 2 epithelial cells (SV40T2). The upregulation of those genes was confirmed by Northern blot analysis and categorized into three groups (highly, moderately, and weakly inducible genes). Heme oxygenase-1 (HO-1), HSP 72, hepatic steroid hydroxylase/CYPIIA2, and Cd inducible gene 1 (cdig1, a new gene, Accession Nos. AB086233 and AB086234) were highly inducible genes, testosterone-repressed prostate message 2 mRNA was moderately inducible, and collagen-binding protein and cdig2 (another new gene, Accession No. AB086193) mRNAs were weakly inducible. The expression of cdig1 increased linearly with time up to 9h, while that of HO-1 reached the maximum value at 4h in response to 10 microM Cd. PMID- 12890492 TI - Identification of a short PIASx gene promoter that directs male germ cell specific transcription in vivo. AB - PIASx gene encodes two SUMO E3 ligases that are highly expressed in the testis. We have isolated and analyzed the promoter of the murine PIASx gene. Electrophoretic mobility shift assays with testicular nuclear extracts showed that the proximal promoter forms a major DNA-protein complex containing Sp1, Sp2, and Sp3 transcription factors. Reporter gene assays in cultured cells indicated that a fragment comprising nucleotides from -168 to +76 relative to transcription start site is sufficient for basal promoter activity in cultured cells, but these in vitro assays failed to reveal clear differences in promoter activity between testis- and non-testis-derived cell lines. Interestingly, however, the proximal promoter encompasses the elements necessary for a testis-specific transcription in vivo, as it directed beta-galactosidase expression exclusively to male germ cells in transgenic mice. In conclusion, we have characterized the minimal PIASx promoter that can be used for highly specific targeting of transgene expression to male germ cells. PMID- 12890493 TI - Binding mechanism of coronavirus main proteinase with ligands and its implication to drug design against SARS. AB - In order to stimulate the development of drugs against severe acute respiratory syndrome (SARS), based on the atomic coordinates of the SARS coronavirus main proteinase determined recently [Science 13 (May) (2003) (online)], studies of docking KZ7088 (a derivative of AG7088) and the AVLQSGFR octapeptide to the enzyme were conducted. It has been observed that both the above compounds interact with the active site of the SARS enzyme through six hydrogen bonds. Also, a clear definition of the binding pocket for KZ7088 has been presented. These findings may provide a solid basis for subsite analysis and mutagenesis relative to rational design of highly selective inhibitors for therapeutic application. Meanwhile, the idea of how to develop inhibitors of the SARS enzyme based on the knowledge of its own peptide substrates (the so-called "distorted key" approach) was also briefly elucidated. PMID- 12890494 TI - Shear stress-induced collagen XII expression is associated with atherogenesis. AB - Fluid shear stress has been shown to modulate various endothelial functions. We selected a shear stress-specific clone, identified as collagen XII, from a bovine aortic endothelial cell (BAEC) cDNA library. We confirmed that shear stress induces collagen XII expression at both the mRNA and protein levels in cultured BAECs and human umbilical vein ECs (HUVECs) by stimulating transcription. When HUVECs were exposed to shear stress, they secreted collagen XII protein and it was deposited underneath them. Strong expression of collagen XII was found in the intima of human aortic wall lacking atherosclerotic lesions, whereas weak expression was seen in the intima of atherosclerotic plagues. Furthermore, the downstream portion of atherosclerotic plaques showed apparently weak collagen XII expression compared with the upstream portion. These results suggest that collagen XII expression induced by fluid shear stress may play a role in stabilizing the vascular structure and preventing the formation of atherosclerotic lesions. PMID- 12890495 TI - Myosin-Va proteolysis by Ca2+/calpain in depolarized nerve endings from rat brain. AB - Myosin-Va is a molecular motor that may participate in synaptic vesicle cycling. Calpain cleaves myosin-Va in vitro at methionine 1141 in the tail domain. We show that intracellular proteolysis of myosin-Va occurs in rat cortical synaptosomes depolarized in the presence of calcium, evidenced by the formation of an 80 k polypeptide that co-migrates in SDS-PAGE with the 80 k fragment produced by the in vitro proteolysis of myosin-Va by calpain. Anti-myosin-Va antibody recognized this polypeptide in Western blots and immunoprecipitated it from synaptosome extracts. Calpastatin, a calpain-specific inhibitor, or leupeptin, a general cysteine protease inhibitor, suppressed or blocked formation of the 80 k polypeptide depending on membrane permeability. We conclude that myosin-Va undergoes intracellular proteolysis by endogenous calpain, when synaptosomes are depolarized in the presence of calcium, at the same cleavage site previously identified in vitro, thus, making it a target for calcium signaling during synaptic activation. PMID- 12890496 TI - Surface trapping and AFM detection of DNA topological intermediates generated from an oxidative chemical nuclease. AB - Direct observation of DNA topological intermediates generated from a 'chemical nuclease' treatment has been made by atomic force microscopy (AFM). The intermediates were trapped at the mica-water interface and imaging was carried out in the dynamic force mode. Complete conversion from supercoiled circular state to relaxed circular/linear state has been observed over a time scale of 8 min. Implication of such studies in complementing gel electrophoresis data has been predicted. PMID- 12890497 TI - CHD1 associates with NCoR and histone deacetylase as well as with RNA splicing proteins. AB - CHD1 is one of a family of nuclear proteins containing two chromodomains, a SWI/SNF-like helicase/ATPase domain and a DNA binding domain. We found that CHD1 co-immunoprecipitates with histone deacetylase (HDAC) activity and that CHD1 also associates with NCoR, a transcriptional corepressor, in yeast two-hybrid and in vitro pull-down assays. NCoR is known to associate with HDACs to effect its repressive activity, suggesting that the predicted chromatin remodeling activity of CHD1 plays a role in this repression. Yeast two-hybrid assays also showed that CHD1 interacts with splicing proteins mKIAA0164, Srp20, and SAF-B. Splicing assays show that CHD1 overexpression can affect alternative splicing. These results suggest that CHD1 may function in both chromatin mediated transcriptional repression and RNA splicing. PMID- 12890498 TI - AT1A-deficient mice show less severe progression of liver fibrosis induced by CCl(4). AB - The renin-angiotensin system has been shown to contribute to fibrogenesis in varieties of organs, including the liver. Here, we investigated whether the angiotensin II type 1A receptor (AT1A) is implicated in the development of liver fibrosis, using AT1A-deficient and wild-type (WT) mice. After single dose of carbon tetrachloride (CCl(4)), there were no significant differences between two groups with regard to hepatic inflammation and necrosis. After 4 weeks of treatment with CCl(4), histological examination revealed that AT1A-deficient mice showed less infiltration of inflammatory cells and less severe progression of liver fibrosis compared with WT mice. These findings were accompanied by the hepatic content of hydoxyproline and the expression of alpha-smooth muscle actin (alpha SMA). The level of transforming growth factor-beta 1 (TGF-beta 1) messenger RNA was markedly higher in WT mice when compared with AT1A-deficient mice. These results confirm that signaling via AT1A plays a pivotal role in hepatic fibrogenesis. PMID- 12890499 TI - Zath3, a neural basic helix-loop-helix gene, regulates early neurogenesis in the zebrafish. AB - We have isolated a basic helix-loop-helix (bHLH) gene homologous to the Drosophila proneural gene atonal, termed zath3, from zebrafish. zath3 is expressed in neurons of the central nervous system and in subsets of cranial ganglia. Zebrafish mindbomb (mib) mutants have a higher density of zath3 expressing cells and narrowminded (nrd) mutants lack zath3 expression in a domain corresponding to primary sensory neurons showing that the expression of zath3 is regulated by both mib and nrd. Injection of synthetic zath3 RNA into zebrafish embryos expands the neural plate size, promotes ectopic expression of neuronal markers, and partially rescues the deficit of sensory neurons seen in nrd mutants. Interfering with zath3 function using antisense morpholino oligonucleotides (MO) has no significant effect on early neurogenesis. However, a double knock down of zath3 and neurogenin1 (ngn1), another atonal homologue, with morpholinos (MOs) leads to more severe defects in neurogenesis than are seen with ngn1 MO alone: a subtle reduction of motor and inter-neurons, and an almost complete loss all cranial ganglia. This study suggests that zath3 and ngn1 have partially overlapping roles in early neurogenesis. PMID- 12890500 TI - A novel method for viral display of ER membrane proteins on budded baculovirus. AB - The baculovirus expression system has been used to express large quantities of various proteins, including membrane receptors. Here, we reveal a novel property of this expression system to be that certain membrane proteins can be displayed on the budded virus itself. We introduced the genes encoding sterol regulatory element-binding protein-2 (SREBP-2) or SREBP cleavage-activating protein (SCAP), important integral membrane proteins of the endoplasmic reticulum (ER) and/or the Golgi apparatus related to cellular cholesterol regulation, into a baculovirus vector. When insect cells were infected with SREBP-2 or SCAP recombinant viruses, it was found that these ER membrane proteins appeared on the budded baculovirus in addition to the host cell membrane fraction. Compared to proteins expressed on the cell membrane, membrane proteins displayed on virus exhibited both less aggregation and less degradation upon immunoblotting. Using this viral displayed SCAP as the screening antigen, we then generated a new monoclonal antibody specific against SCAP, which was useful for immunological localization studies. This system, which takes advantage of the viral display of membrane proteins, should prove to be a powerful additional tool for postgenomic protein analysis. PMID- 12890501 TI - Profiling genes related to mitochondrial function in mice treated with N-methyl-4 phenyl-1,2,3,6-tetrahydropyridine. AB - Since mitochondrial dysfunction plays an important role in the pathogenesis of dopaminergic neurodegeneration in Parkinson's disease, we determined the expression of genes related to mitochondrial function in the substantia nigra of mice treated with N-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) using a cDNA array. MPTP treatment significantly depleted striatal dopamine, but did not result in apparent neuronal loss in the substantia nigra at 3 and 18 days post treatment. We also examined changes in genes in the hypothalamus, a region containing dopaminergic neurons that are relatively resistant to MPTP. Finally, we confirmed those genes identified by microarrays as differentially expressed in the substantia nigra but not in the hypothalamus using in situ hybridization. Our results demonstrated that MPTP significantly changed the expressions of six genes in nigral neurons, four of which were related to the mitochondrial electron transport chain: the NADH-ubiquinone oxidoreductase 13 kDa B subunit, the NADH ubiquinone oxidoreductase MNLL subunit, cytochrome c, and the cytochrome c oxidase Va subunit. Two other differentially expressed genes were the dihydropyridine-sensitive L-type calcium channel alpha-2 subunit precursor and type III alpha-1 procollagen. None of these six genes are encoded by mitochondrial DNA. The potential significance of these gene alterations in the context of Parkinson's disease is discussed. PMID- 12890503 TI - Gene regulation of melatonin and dopamine receptors during eye development. AB - To gain insight into the role of melatonin and dopamine in retinal development, gene expression of two melatonin receptors, MT1 and MT2, as well as five dopamine receptors, D1, D2, D3, D4 and D5, in the rat eye was analyzed by reverse transcription-polymerase chain reaction across various developmental stages. MT1 transcript levels reached maximum levels at embryonic day (E) 16 and then decreased gradually until reaching adult levels by postnatal day (P) 14. MT2 transcript levels similarly peaked at E16, but then decreased dramatically until birth to its lowest levels, which were maintained throughout the postnatal period. Thus, gene expression of both the MT1 and MT2 receptors showed a striking inverse correlation with maturation of the eye. In contrast to melatonin receptors, gene expression of all dopamine receptor subtypes, except for D3, showed only an increase as development proceeds with highest levels in adulthood. The D3 message was not detected throughout the developmental period examined. Gene expression of D1-like receptors, D1 and D5, showed a substantial increase to adult levels during the fetal period at E16 and E20, respectively. Transcript levels of D2-like receptors, D2 and D4, on the other hand, were not detected before birth but increased significantly to adult levels by P7 and P14, respectively. The present findings suggest the presence of unique developmental mechanisms by which transcription of various G protein-coupled receptors are regulated in the eye. PMID- 12890502 TI - Identification of neuronal plasma membrane microdomains that colocalize beta amyloid and presenilin: implications for beta-amyloid precursor protein processing. AB - Alzheimer's disease (AD) is associated with the accumulation of extracellular deposits of the beta-amyloid protein (Abeta). Abeta is a result of misprocessing of the beta-amyloid precursor protein (APP). Gamma-secretase is involved in APP misprocessing and one hypothesis holds that this secretase is identical to PS1. We tested this hypothesis by determining whether PS is co-localised with Abeta in situ. Using confocal analyses and a sensitive immunogold procedure we show that PS and Abeta are co-localised within discrete microdomains of neuronal plasma membranes in AD patients and in aged dogs, an established model of human brain aging. Our data indicate that APP misprocessing occurs in discrete plasma membrane domains of neurons and provide evidence that PS1 is critically involved in Abeta formation. PMID- 12890504 TI - The distribution of the orphan bombesin receptor subtype-3 in the rat CNS. AB - Bombesin receptor subtype 3 (BRS-3) is an orphan G-protein coupled receptor that shares between 47 and 51% homology with other known bombesin receptors. The natural ligand for BRS-3 is currently unknown and little is known about the mechanisms regulating BRS-3 gene expression. Unlike other mammalian bombesin receptors that have been shown to be predominantly expressed in the CNS and gastrointestinal tract, expression of the BRS-3 receptor in the rat brain has previously not been observed. To gain further understanding of the biology of BRS 3, we have studied the distribution of BRS-3 mRNA and protein in the rat CNS. The mRNA expression pattern was studied using reverse transcription followed by quantitative polymerase chain reaction. Using immunohistological techniques, the distribution of BRS-3 protein in the rat brain was investigated using a rabbit affinity-purified polyclonal antiserum raised against an N-terminal peptide. The BRS-3 receptor was found to be widely expressed in the rat brain at both mRNA and protein levels. Particularly strong immunosignals were observed in the cerebral cortex, hippocampal formation, hypothalamus and thalamus. Other regions of the brain such as the basal ganglia, midbrain and reticular formation were also immunopositive for BRS-3. In conclusion, our neuroanatomical data provide evidence that BRS-3 is as widely expressed in the rat brain as other bombesin like peptide receptors and suggest that this receptor may also have important roles in the CNS, mediating the functions of a so far unidentified ligand. PMID- 12890505 TI - Effect of anti-nerve growth factor treatment on pituitary adenylate cyclase activating polypeptide expression in adult sensory neurons exposed to adjuvant induced inflammation. AB - Expression of pituitary adenylate cyclase activating polypeptide (PACAP) is increased in sensory neurons exposed to adjuvant induced peripheral inflammation. Local elevation in expression of the neurotrophin nerve growth factor (NGF) is a main factor contributing to the neuronal response to inflammation. This study examines the role of endogenous NGF in inflammation-associated increases in PACAP expression using the adjuvant-induced peripheral inflammation model with or without systemic administration of antibodies against NGF. Quantitative in situ hybridization was used to detect changes in neuronal PACAP mRNA expression and to correlate this expression with neuronal mRNA expression of the NGF receptor tyrosine kinase (trk) A. The results from this study show that inflammation triggered increases in PACAP expression occurs in small- to medium-sized dorsal root ganglion (DRG) neurons that also express trkA, and that this elevation in PACAP expression is prevented by systemic injection of anti-NGF. This supports a role for NGF as a positive regulator of PACAP expression during inflammation. PMID- 12890506 TI - Inhibitory cotransmission or after-hyperpolarizing potentials can regulate firing in recurrent networks with excitatory metabotropic transmission. AB - Recurrent networks of neurons communicating via excitatory connections are common in the nervous system. In the absence of mechanisms to control firing (collectively termed negative feedback), these networks are likely to be bistable and unable to meaningfully encode input signals. In most recurrent circuits, negative feedback is provided by a specialized subpopulation of interneurons, but such neurons are absent from some systems, which therefore require other forms of negative feedback. One such circuit is found within the enteric nervous system of the intestine, where AH/Dogiel type II neurons are interconnected via excitatory synapses acting through metabotropic receptors to produce slow excitatory postsynaptic potentials (slow EPSPs). Negative feedback in this recurrent network may come from either inhibitory postsynaptic potentials arising from the terminals that produce slow EPSPs or from the after hyperpolarizing potentials (AHPs) characteristic of these neurons. We have examined these possibilities using mathematical analysis, based on the Wilson-Cowan model, and computer simulations. Analysis of steady states showed that, under appropriate conditions, both types of negative feedback can provide robust regulation of firing allowing the networks to encode input signals. Numerical simulations were performed using large, anatomically realistic networks with realistic models for metabotropic transmission and suppression of the AHP. In the presence of constant exogenous input, parameters controlling aspects of synaptic events were varied, confirming the analytical results for static stimuli. The simulated networks also responded to time varying inputs in a manner consistent with known physiology. In addition, simulation revealed that neurons in networks with inhibitory contransmission fired in erratic bursts, a phenomenon observed in neurons in unparalysed tissue. Thus, either inhibitory contransmission or AHPs, or both, can allow recurrent networks of AH/Dogiel type II neurons to encode ongoing inputs in a biologically useful way. These neurons appear to be intrinsic primary afferent neurons (IPANs), which implies that the IPANs in a region act in a coordinated fashion. PMID- 12890507 TI - Localization of KCNQ5 in the normal and epileptic human temporal neocortex and hippocampal formation. AB - The KCNQ family of voltage-dependent non-inactivating K+ channels is composed of five members, four of which (KCNQ2-5) are expressed in the CNS and are responsible for the M-current. Mutations in either KCNQ2 or KCNQ3 lead to a hereditary form of dominant generalized epilepsy. Using specific antisera to the KCNQ2, KCNQ3 and KCNQ5 subunits, we found that KCNQ3 co-immunoprecipitated with KCNQ2 and KCNQ5 subunits, but no association was detected between KCNQ2 and KCNQ5. Intense KCNQ5 immunoreactivity was found to be widely distributed throughout the temporal neocortex and the hippocampal formation. In these structures, both pyramidal and non-pyramidal neurons and a population of glial cells in the white matter expressed the KCNQ5 subunit. In the sclerotic areas of the CA fields of epileptic patients, a marked loss of KCNQ5 immunoreactive pyramidal neurons was found in relation with the loss of neurons in these regions. However, in the regions adjacent to the sclerotic areas, the distribution and intensity of KCNQ5 immunostaining was apparently normal. The widespread distribution of KCNQ5 subunits, its persistence in pharmacoresistant epilepsy, along with the significant role of the M-current in the control of neuronal excitability, makes this protein a possible target for the development of anticonvulsant drugs. PMID- 12890508 TI - Acetoacetate protects hippocampal neurons against glutamate-mediated neuronal damage during glycolysis inhibition. AB - Glucose is the main substrate that fulfills energy brain demands. However, in some circumstances, such as diabetes, starvation, during the suckling period and the ketogenic diet, brain uses the ketone bodies, acetoacetate and beta hydroxybutyrate, as energy sources. Ketone body utilization in brain depends directly on its blood concentration, which is normally very low, but increases substantially during the conditions mentioned above. Glutamate neurotoxicity has been implicated in neurodegeneration associated with brain ischemia, hypoglycemia and cerebral trauma, conditions related to energy failure, and to elevation of glutamate extracellular levels in brain. In recent years substantial evidence favoring a close relation between glutamate neurotoxic potentiality and cellular energy levels, has been compiled. We have previously demonstrated that accumulation of extracellular glutamate after inhibition of its transporters, induces neuronal death in vivo during energy impairment induced by glycolysis inhibition. In the present study we have assessed the protective potentiality of the ketone body, acetoacetate, against glutamate-mediated neuronal damage in the hippocampus of rats chronically treated with the glycolysis inhibitor, iodoacetate, and in hippocampal cultured neurons exposed to a toxic concentration of iodoacetate. Results show that acetoacetate efficiently protects against glutamate neurotoxicity both in vivo and in vitro probably by a mechanism involving its role as an energy substrate. PMID- 12890509 TI - The differentiation potential of precursor cells from the mouse lateral ganglionic eminence is restricted by in vitro expansion. AB - We have investigated whether the differentiation potential of attached cultures derived from the mouse lateral ganglionic eminence (LGE) is influenced by in vitro expansion. Primary neuronal cultures derived from the LGE give rise to neurons expressing the striatal projection neuron markers Islet1 (ISL1) and dopamine and cAMP-regulated phosphoprotein of 32 kilodaltons (DARPP-32) as well as the olfactory bulb interneuron marker Er81. Our previous results showed that after expansion in vitro, LGE precursor cells can be induced to differentiate into neurons which exhibit molecular characteristics of the LGE, such as the homeobox transcription factors DLX and MEIS2. We show here that while attached LGE cultures maintain Er81 expression through five passages, they lose the ability to generate ISL1- or dopamine and cAMP-regulated phosphoprotein of 32 kilodaltons-expressing neurons already after the first passage. This indicates that the expansion of LGE precursor cells restricts their differentiation potential in vitro. Interestingly, the undifferentiated LGE cultures retain the expression of both the Isl1 and Er81 genes, suggesting that precursor cells for both striatal projection neurons and olfactory bulb interneurons are present in these cultures. Thus the restriction in differentiation potential of the expanded LGE cultures likely reflects deficiencies in the differentiation conditions used. PMID- 12890510 TI - Developmental regulation of the A-type potassium-channel current in hippocampal neurons: role of the Kvbeta 1.1 subunit. AB - The rapidly inactivating A-type K+ current (IA) is prominent in hippocampal neurons; and the speed of its inactivation may regulate electrical excitability. The auxiliary K+ channel subunit Kvbeta 1.1 confers fast inactivation to Shaker related channels and is postulated to affect IA. Whole-cell patch clamp recordings of rat hippocampal pyramidal neurons in primary culture showed a developmental decrease in the time constant of inactivation (tau(in)) of voltage gated K+ currents: 17.9+/-1.5 ms in young neurons (5-7 days in vitro; n=53, mean+/-S.E.M.); 9.9+/-1.0 ms in mature neurons (12-15 days in vitro; n=72, mean+/ S.E.M., P<0.01). During the same developmental time, the level of Kvbeta 1.1 transcript increased more than two-fold in vitro and in vivo, determined by semi quantitative reverse transcriptase-polymerase chain reaction for hippocampus. The hypothesis that up-regulation of Kvbeta 1.1 led to the changes in tau(in) was tested in vitro, using antisense knockdown. Kvbeta 1.1-specific antisense DNA was introduced with a modified herpes virus co-expressing enhanced green fluorescent protein and knockdown of Kvbeta 1.1 was verified by immunocytochemistry. Following transduction with the antisense virus, mature neurons reverted to tau(in) values characteristic of young neurons: 18.3+/-2.4 ms (n=20). The effect of antisense knockdown on electrical excitability was tested using current-clamp protocols to induce repetitive firing. Treatment with the antisense virus increased the interspike interval over a range of membrane depolarization (baseline membrane potential=-40 to +20 mV). This effect was most pronounced at 40 mV, where the ISI of the first pair of action potentials was nearly doubled. These data indicate that Kvbeta 1.1 contributes to the developmental control of IA in hippocampal neurons and that the magnitude of effect is sufficient to regulate electrical excitability. Viral-mediated antisense knockdown of Kvbeta 1.1 is capable of decreasing the electrical excitability of post-mitotic hippocampal neurons, suggesting this approach has applicability to gene therapy of neurological diseases associated with hyperexcitability. PMID- 12890511 TI - Alzheimer's disease proteins in cerebellar and hippocampal synapses during postnatal development and aging of the rat. AB - Alzheimer's dementia may be considered a synaptic disease of central neurons: the loss of synapses, reflected by early cognitive impairments, precedes the appearance of extra cellular focal deposits of beta-amyloid peptide in the brain of patients. Distinct immunocytochemical patterns of amyloid precursor proteins (APPs) have previously been demonstrated in the synapses by ultrastructural analysis in the cerebellum and hippocampus of adult rats and mice. Now we show that during postnatal development and during aging in these structures, the immunocytochemical expression of APPs increases in the synapses in parallel with the known up-regulation of total APPs brain levels. Interestingly, as shown previously in the adult rodents, the presenilins (PSs) 1 and 2, which intervene in APPs metabolism, exhibit a synaptic distribution pattern similar to that of APPs with parallel quantitative changes throughout life. In the brain tissue, single and double immunocytochemistry at the ultrastructural level shows co localisation of APPs and PSs in axonal and dendritic synaptic compartments during postnatal synaptogenesis, adulthood and aging. In addition, double-labelling immunocytofluorescence detects these proteins close to synaptophysin at the growth cones of developing cultured neurons. Thusly, the brain expression of APPs and PSs appears to be regulated synchronously during lifespan in the synaptic compartments where the proteins are colocated. This suggests that PS-dependent processing of important synaptic proteins such as APPs could intervene in age induced adjustments of synaptic relationships between specific types of neurons. PMID- 12890512 TI - An investigation of dopaminergic metabolites in the striatum and in the substantia nigra in vivo utilising radiolabelled L-DOPA and high performance liquid chromatography: a new approach in the search for transmitter metabolites. AB - Although the major routes of dopamine metabolism seem to be established, at least in terminal regions such as the striatum, it is important to search for previously unknown metabolites and to investigate the relevance of previously suggested minor alternative pathways. An urgent issue is to verify and quantify the transformation of dopamine to putative toxic species, another is to further explore metabolism of dopamine located in cell bodies/dendrites, e.g. in the substantia nigra. We have developed a new method in order to widen the search for alternative metabolites of dopamine. The method is based on systemic injection of tritiated L-DOPA to rats in vivo. Brain tissue was homogenised and centrifuged and the resulting supernatant fractioned following passage through a liquid chromatography system. The radioactivity of each fraction was measured using a scintillation system. By identifying fractions containing major catecholamines and metabolites, according to a standard solution, novel metabolites can be searched for in the remaining fractions. It was possible to obtain sufficient radioactivity in separate fractions of supernatant of homogenised tissue, even from such a small brain nucleus as substantia nigra. Radioactivity was obtained in those fractions that contained the major catecholamines and their metabolites, as well as in other fractions where it may represent previously unknown metabolites of L-DOPA/dopamine. The method was used to evaluate the possibility that cytochrome P450 2E1 is involved in the metabolism of dopamine in the substantia nigra. Significant changes in the radioactivity pattern were induced by inhibition of the enzyme but conclusions about whether cytochrome P450 2E1 is involved in the metabolism of dopamine or not requires further studies. The method can be used to study the metabolism of dopamine and can be extended, by using other radiolabelled precursors, also to evaluate metabolism of other transmitters, e.g. serotonin. PMID- 12890513 TI - Neurological phenotype and synaptic function in mice lacking the CaV1.3 alpha subunit of neuronal L-type voltage-dependent Ca2+ channels. AB - Neuronal L-type calcium channels have been implicated in pain perception and neuronal synaptic plasticity. To investigate this we have examined the effect of disrupting the gene encoding the CaV1.3 (alpha 1D) alpha subunit of L-type Ca2+ channels on neurological function, acute nociceptive behavior, and hippocampal synaptic function in mice. CaV1.3 alpha 1 subunit knockout (CaV1.3 alpha 1(-/-)) mice had relatively normal neurological function with the exception of reduced auditory evoked behavioral responses and lower body weight. Baseline thermal and mechanical thresholds were unaltered in these animals. CaV1.3 alpha 1(-/-) mice were also examined for differences in N-methyl-D-aspartate (NMDA) receptor dependent (100 Hz tetanization for 1 s) and NMDA receptor-independent (200 Hz in 100 microM DL-2-amino-5-phosphopentanoic acid) long-term potentiation within the CA1 region of the hippocampus. Both NMDA receptor-dependent and NMDA receptor independent forms of long-term potentiation were expressed normally. Radioligand binding studies revealed that the density of (+)[3H]isradipine binding sites in brain homogenates was reduced by 20-25% in CaV1.3 alpha 1(-/-) mice, without any detectable change in CaV1.2 (alpha 1C) protein levels as detected using Western blot analysis. Taken together these data indicate that following loss of CaV1.3 alpha 1 subunit expression there is sufficient residual activity of other Ca2+ channel subtypes to support NMDA receptor-independent long-term potentiation and some forms of sensory behavior/function. PMID- 12890514 TI - Growth hormone secretagogue receptors in rat and human gastrointestinal tract and the effects of ghrelin. AB - The peptide hormone ghrelin is known to be present within stomach and, to a lesser extent, elsewhere in gut. Although reports suggest that gastric function may be modulated by ghrelin acting via the vagus nerve, the gastrointestinal distribution and functions of its receptor, the growth hormone secretagogue receptor (GHS-R), are not clear and may show signs of species-dependency. This study sought to determine the cellular localisation and distribution of GHS-R immunoreactivity (-Ir) using immunofluorescent histochemistry and explore the function of ghrelin in both human and rat isolated gastric and/or colonic circular muscle preparations in which nerve-mediated responses were evoked by electrical field stimulation. The expression of GHS-R-Ir differed to a greater extent between species than between gut regions of the same species. Both the human and rat gastric and colonic preparations (n=3 each) expressed GHS-R-Ir within neuronal cell bodies and fibres, cells associated with gastric glands and putative entero-endocrine and/or mast cells. Smooth muscle cells and epithelia were devoid of GHS-R-Ir and only rat preparations expressed GHS-R-Ir on nerve fibres associated with the muscle layers. GHS-R-Ir was fully competed in all cases in pre-adsorption studies and antiserum specificity was confirmed using a cell line transiently expressing the rat GHS-R. In rat isolated forestomach circular muscle, ghrelin 0.1-10 microM had no effect on smooth muscle tension but concentration-dependently facilitated the amplitude of contractions evoked by excitatory nerve stimulation (n=4-7; P<0.05 for each concentration versus vehicle; n=18). When examined under similar conditions, in both rat distal colon (n=4-6, P>0.05 each) and human ascending (n=3) and sigmoid (n=1) colon, these concentrations of ghrelin were without effect (P>0.05 each). The data suggest that ghrelin has the potential to profoundly affect gastrointestinal functions in both species and at least one of these functions is to exert a gastric prokinetic activity. Moreover, we suggest that this activity of ghrelin is mediated via the enteric nervous system, in addition to known vagus nerve-dependent mechanisms. PMID- 12890515 TI - Modulation of electrically evoked acetylcholine release through cannabinoid CB1 receptors: evidence for an endocannabinoid tone in the human neocortex. AB - Cannabinoids are known to inhibit neurotransmitter release in the CNS through CB1 receptors. The present study compares the effects of synthetic cannabinoids on acetylcholine (ACh) release in human and mice neocortex. We further investigated a possible endocannabinoid tone on CB1 receptors in human neocortex caused by endogenous agonists like anandamide or 2-arachidonylglycerol. Brain slices, incubated with [3H]-choline, were superfused and stimulated electrically under autoinhibition-free conditions to evoke tritium overflow assumed to represent ACh release. The first series of experiments was performed with 26 pulses, 60 mA, at 0.1 Hz. In mice neocortical slices, the cannabinoid receptor agonist WIN55212-2 decreased ACh release (pIC50=6.68, I(max)=67%). In the human neocortex the concentration-response curve of WIN55212-2 was bell-shaped and flat (I(max observed) approximately 30%). The estimated maximum possible inhibition, however, was much larger: I(max derived)=79%. Lec, the negative logarithm (lg) of the biophase concentration of endocannabinoids in 'WIN55212-2 units,' was -6.52, the pKd of WIN55212-2 was 7.47. The CB1 receptor antagonist/inverse agonist SR141716 enhanced ACh release in the human neocortex (by 38%) and prevented the inhibitory effect of WIN55212-2. The concentration-response curve of WIN55212-2 was changed in its shape including a shift to the right due to the presence of SR141716. A pA2 of this antagonist between 11.60 and 11.18 was obtained. SR141716 alone had no effect in mice neocortical slices. A partial agonist without inverse agonistic activity, O-1184, enhanced ACh release in the human neocortex. The endocannabinoid uptake-inhibitor AM404 decreased ACh release in human, but not in mice, neocortical slices. Change of the stimulation parameters (eight trains of pseudo-one-pulse bursts (4 pulses, 76 mA, 100 Hz), spaced by 45 s intervals) led to a stronger inhibitory effect of WIN55212-2, and abolished the disinhibitory effect of SR141716 and O-1184. The results show that activation of CB1 cannabinoid receptors leads to inhibition of ACh release in the human and mouse neocortex. The endocannabinoid tone is high in the human, but not in the mouse neocortex and is dependent on neuronal activity. SR141716 acts as a competitive CB1 receptor antagonist. PMID- 12890516 TI - Pro-nociceptive effects of neuromedin U in rat. AB - The neuropeptide neuromedin U (NMU) has been shown to have significant effects on cardiovascular, gastrointestinal and CNS functions. The peptide was first isolated from the porcine spinal cord and later shown to be present in spinal cords of other species. Little is known about the distribution of neuromedin U receptors (NMURs) in the spinal cord and the spinal action of the peptide. Here we report on the expression of NMURs and a potential role in nociception in the rat spinal cord using a combination of behavioral and electrophysiological studies. Receptor autoradiography showed that NMU-23 binding was restricted to the superficial layers of spinal cord, a region known to be involved in the control of nociception. In situ hybridization analysis indicated the mRNA of NMUR2 was located in the same region (laminae I and IIo) as NMU-23 binding, while the mRNA for NMU receptor 1 was observed in a subpopulation of small diameter neurons of dorsal root ganglia. Intrathecal (i.t.) administration of neuromedin U 23 (0.4-4.0 nmol/10 microl) dose-dependently decreased both the mechanical threshold to von Frey hair stimulation and the withdrawal latency to a noxious thermal stimulus. Mechanical allodynia was observed between 10 and 120 min, peaking at 30 min and heat hyperalgesia was observed 10-30 min after i.t. administration of NMU-23. A similar mechanical allodynia was also observed following i.t. administration of NMU-8 (0.4-4 nmol/10 microl). A significant enhancement of the excitability of flexor reflex was induced by intrathecal administration of NMU-23 (4 nmol/10 microl). Evoked responses to touch and pinch stimuli were increased by 439+/-94% and 188+/-36% (P<0.01, n=6) respectively. The behavioral and electrophysiological data demonstrate, for the first time, a pro nociceptive action of NMU. The restricted distribution of NMU receptors to a region of the spinal cord involved in nociception suggests that this peptide receptor system may play a role in nociception. PMID- 12890517 TI - Neuroprotective activity of antazoline against neuronal damage induced by limbic status epilepticus. AB - Imidazoline drugs exert neuroprotective effects in cerebral ischaemia models. They also have effects against mouse cerebellar and striatal neuronal death induced by N-methyl-D-aspartate (NMDA) through the blockade of NMDA currents. Here, we investigated the effects of antazoline on NMDA toxicity and current in rat hippocampal neuronal cultures, and on an in vivo model of status epilepticus. In hippocampal cultures, antazoline (30 microM) decreased NMDA-mediated neurotoxicity and also blocked the NMDA current with voltage-dependent and fast reversible action (inhibition by 85+/-3% at -60 mV). Status epilepticus was induced by injecting pilocarpine (200 nmol) directly into the right pyriform cortex of male adult rats. The rats then received immediately three consecutive i.p. injections at 30-min intervals of either PBS (control group) or antazoline at 10 mg/kg (low-dose group) or at 45 mg/kg (high-dose group). During the 6-h recording, status epilepticus lasted more than 200 min in all groups. In the high dose group only, seizures completely ceased 1 h after the third injection of antazoline, then started again 1 h later. Rats were killed 1 week later, and Cresyl Violet-stained sections of their brain were analysed for damage quantification. On the ipsilateral side to the pilocarpine injection, pyriform cortex and hippocampal CA1 and CA3 areas were significantly protected in both antazoline-treated groups, whilst prepyriform and entorhinal cortices were only in the high-dose group. On the contralateral side to the pilocarpine injection, only the hippocampal CA3 area was significantly protected in the low-dose group, but all investigated structures were in the high-dose group. In conclusion, antazoline is a potent neuroprotective drug in different models of neuronal primary culture, as previously shown in striatal and cerebellar granule neurons [Neuropharmacology 39 (2000) 2244], and here in hippocampal neurons. Antazoline is also neuroprotective in vivo in the intra-pyriform pilocarpine-induced status epilepticus model. PMID- 12890518 TI - Brain afferents to the lateral caudal ventrolateral medulla: a retrograde and anterograde tracing study in the rat. AB - The ventrolateral medulla (VLM) modulates autonomic functions, motor reactions and pain responses. The lateralmost part of the caudal VLM (VLMlat) was recently shown to be the VLM area responsible for pain modulation. In the present study, the brain sources of VLMlat afferent fibers were determined by tract-tracing techniques. Following injection of cholera toxin subunit B into the VLMlat, retrogradely labeled neurons in the forebrain occurred at the somatosensory, insular, motor, limbic and infralimbic cortices, and at the central amygdaloid nucleus. Retrogradely labeled neurons in diencephalic regions were observed in the lateral hypothalamus, posterior hypothalamus and paraventricular nucleus. In the brainstem, retrograde labeling occurred at the periaqueductal gray, red nucleus, parabrachial area, nucleus raphe magnus, nucleus tractus solitarii, lateral reticular nucleus and dorsal and ventral medullary reticular formation. In the cerebellum, retrogradely labeled neurons occurred at the lateral nucleus. Following injections of the anterograde tracer biotinylated dextran amine (BDA) into the lateral hypothalamus or paraventricular nucleus, anterogradely labeled fibers were mainly observed in the VLMlat. Injections of BDA into the periaqueductal gray, red nucleus or lateral nucleus of the cerebellum resulted in anterograde labeling in the VLMlat and lateral reticular nucleus. The present study gives an account of the brain regions putatively involved in triggering the modulatory actions elicited from the VLMlat. These include areas committed to somatosensory processing, autonomic control, somatic and visceral motor activity and affective reactions. The findings suggest that the VLMlat may play a major homeostatic role in the integration of nociception with other brain functions. PMID- 12890519 TI - A role for peripheral somatostatin receptors in counter-irritation-induced analgesia. AB - Our hypothesis is that peripheral somatostatin (SRIF) has a role in counter irritation-induced analgesia. Our paradigm involves the reduction of nociceptive behaviors produced by primary noxious stimuli (formalin or complete Freund's adjuvant [CFA] in the rat hind paw) by a counter-irritating stimulus (capsaicin [CAP] in the tail or muzzle). Activation of peripheral SRIF receptors is key since an SRIF receptor antagonist cyclo-somatostatin (c-SOM) and SRIF antibodies in the hind paw attenuate the counter-irritation-induced analgesia of both formalin and more persistent CFA nociception. Specificity of c-SOM is shown by reversal of its effects with octreotide, a SRIF analog. Injection of formalin in one hind paw and c-SOM in the other does not reduce the counter-irritation analgesia demonstrating local action of the c-SOM. Approximately 33% of peripheral sensory axons contain SRIF, which could release the peptide to activate SRIF receptors on cutaneous axons. Intraplantar naloxone has no effect on the counter-irritation analgesia indicating that SRIF is not activating opioid receptors. These results indicate that in addition to the classic central descending noxious inhibitory control systems that underlie counter-irritation induced analgesia, there is a peripheral contribution arising from activation of SRIF receptors. Identifying a peripheral contribution of SRIF to mechanisms of counter-irritation analgesia offers opportunities for peripheral therapy. PMID- 12890520 TI - Modulation of theta rhythmicity in the medial septal neurons and the hippocampal electroencephalogram in the awake rabbit via actions at noradrenergic alpha2 receptors. AB - The modulation of the firing discharge of medial septal neurons and of the hippocampal electroencephalogram (EEG) mediated by actions on alpha2 adrenoreceptors (ARs) was investigated in awake rabbits. Bilateral i.c.v. infusion of a relatively low dose (0.5 microg) of the alpha2-AR agonist clonidine produced a reduction in the theta rhythmicity of both medial septal neurons and the hippocampal EEG. In contrast, a high dose of clonidine (5 microg) increased the percentage and degree of rhythmicity of theta bursting medial septal neurons as well as the theta power of the hippocampal EEG. On the other hand, administration of alpha2-AR antagonist idazoxan produced the opposite dose dependent effect. While a low dose of the antagonist (20 microg) produced an increase in both the theta rhythmicity of medial septal neurons and the theta power of the hippocampal EEG, a high dose (100 microg) caused a reduction of theta rhythmicity in both the medial septum and hippocampus. These results suggest that low doses of alpha2-ARs agents may act at autoreceptors regulating the synaptic release of noradrenaline, while high doses of alpha2-ARs drugs may have a predominant postsynaptic action. Similar results were observed after local injection of the alpha2-AR drugs into the medial septum suggesting that the effects induced by the i.c.v. infusion were primarily mediated at the medial septal level. We suggest that noradrenergic transmission via the postsynaptic alpha2-ARs produces fast and strong activation of the septohippocampal system in situations that require urgent selective attention to functionally significant information (alert, aware), whereas the action via the presynaptic alpha2-ARs allows a quick return of the activity to the initial level. PMID- 12890521 TI - Gonadal hormones differentially modulate cocaine-induced conditioned place preference in male and female rats. AB - There is accumulating evidence that suggests there are sex differences in behavioral and subjective responses to cocaine. However, it is not known whether differences in cocaine reward contribute to sex differences in these responses or whether gonadal hormones affect the rewarding properties of cocaine. In the present study, conditioned place preference (CPP), a measure of non-contingent reward, was used to determine the effects of endogenous gonadal hormones and of estrogen and progesterone replacement on cocaine reward. Neurochemical measurements were also taken to identify monoaminergic substrates which underlie the behavioral phenotype. Although both intact and gonadectomized male and female rats showed a significant CPP for cocaine, ovariectomy attenuated the magnitude of CPP. These alterations coincided with a decrease in serum levels of corticosterone. In ovariectomized rats, pretreatment with progesterone inhibited cocaine CPP while estrogen plus progesterone potentiated the magnitude of CPP. Additionally, gonadectomy and ovarian hormone replacement in female rats affected serotonin/dopamine levels and turnover ratios in the ventral tegmental area and nucleus accumbens shell. While no effects of castration were observed, ovariectomy decreased levels of dopamine and serotonin in the ventral tegmental area. In females, progesterone replacement increased levels of serotonin and dopamine in the ventral tegmental area, while estrogen plus progesterone replacement increased dopamine levels in the nucleus accumbens. Collectively, these results indicate that ovarian hormones may influence cocaine reward by altering monoaminergic systems, which, in turn, may contribute to the current sex disparities in overall cocaine use. PMID- 12890522 TI - Impaired long-term potentiation in obese zucker rats: possible involvement of presynaptic mechanism. AB - Electrophysiological investigation of basal synaptic transmission and synaptic plasticity in the CA1 region of the hippocampus was carried out in anesthetized obese Zucker rats (OZR). Comparison of the input/output curves of basal field excitatory postsynaptic potential indicates that these are similar in both the OZR and its lean counterpart suggesting that basal synaptic transmission is intact in the OZR. However, high frequency stimulation evokes long-term potentiation (LTP) in the lean rat but not in the OZR. Since post-tetanic potentiation and paired pulse facilitation, forms of short-term potentiation of presynaptic origin, are also severely impaired in the OZR, the results imply that impairment of CA1 hippocampal LTP in these obese rats may be due, in part, to impaired presynaptic function. The results emphasize the potential deleterious effect of obesity on learning and memory functions of the CNS. PMID- 12890523 TI - Gender differences in the regulation of 3 alpha-hydroxysteroid dehydrogenase in rat brain and sensitivity to neurosteroid-mediated stress protection. AB - The enzyme 3 alpha-hydroxysteroid dehydrogenase (3 alpha-HSD) is involved in the generation of neuroactive steroids through ring-A-reduction of hormonal precursors. We examined the developmental regulation of, gender differences in, and effects of hormonal manipulations on the expression of 3 alpha-HSD in the rat hippocampus. High levels of 3 alpha-HSD mRNA were found on postnatal day 7, coinciding with the stress hyporesponsive period in the rat. Gender differences in 3 alpha-HSD expression were documented during puberty, but not in adulthood. Adrenalectomy and gonadectomy, and supplementation with individual steroid hormones influenced 3 alpha-HSD expression in a gender-specific mode. We also demonstrate that the manifestation of behavioral and endocrine consequences of early life stress depends on the individual's gender and gonadal status. Males are liable to aftereffects of neonatal maternal deprivation, regardless of their adult gonadal status. In females, however, anxiogenic aftereffects of neonatal stress become apparent only after gonadectomy. These data suggest that (i) transient increase of neurosteroid biosynthesis may contribute to stress hyporesponsiveness during early infancy; (ii) gonadal steroids regulate 3 alpha HSD expression in the hippocampus in a sex-specific mode; (iii) physiological sex steroid secretions in females may mask behavioral consequences of adverse early life events, and (iv) concomitant treatment with the neurosteroid THP counteracts behavioral and endocrine dysregulation induced by neonatal stress in both genders. PMID- 12890524 TI - Environmental context and drug history modulate amphetamine-induced c-fos mRNA expression in the basal ganglia, central extended amygdala, and associated limbic forebrain. AB - The context in which amphetamine is administered modulates its ability to induce both behavioral sensitization and immediate early gene expression. When given in a novel test environment amphetamine produces greater levels of c-fos and arc mRNA expression in many brain regions relative to when it is given in the home cage. The purpose of the current study was to determine if environment and drug history interact to influence amphetamine-induced c-fos mRNA expression. Rats with a unilateral 6-hydroxydopamine lesion were treated for 7 days with saline or 0.5 mg/kg of d-amphetamine (i.v.) in a distinct and relatively novel test environment (Novel), or in their home cage (Home). Following a 10-12-day withdrawal period, a challenge injection of either saline or 0.5 mg/kg d amphetamine was administered. In situ hybridization histochemistry was used to examine c-fos mRNA expression in several regions of the basal ganglia, the central extended amygdala, and limbic forebrain. In most brain regions amphetamine given in the Novel environment produced greater c-fos mRNA expression than when given it was given at Home, and drug history had no effect on amphetamine-induced c-fos mRNA expression. However, within the subthalamic nucleus, substantia nigra reticulata, and central nucleus of the amygdala prior experience with amphetamine in the Novel but not Home environment enhanced the effect of an amphetamine challenge injection on c-fos mRNA expression. In contrast, there was a decrease in c-fos mRNA expression in amphetamine-pretreated animals, regardless of environmental context, in the ventral portion of the far caudal striatum. Reexposure to an environment previously paired with amphetamine produced a conditioned increase in c-fos mRNA expression in portions of the caudate-putamen, the subthalamic nucleus, the nucleus accumbens shell and a conditioned decrease in c-fos mRNA expression in the central nucleus of the amygdala. We conclude that environmental context and drug history interact to alter the basal ganglia and central extended amygdala circuitry engaged by subsequent exposure to amphetamine, or exposure to an environment previously paired with amphetamine. PMID- 12890525 TI - Organization of projections from the raphe nuclei to the vestibular nuclei in rats. AB - Previous anatomic and electrophysiological evidence suggests that serotonin modulates processing in the vestibular nuclei. This study examined the organization of projections from serotonergic raphe nuclei to the vestibular nuclei in rats. The distribution of serotonergic axons in the vestibular nuclei was visualized immunohistochemically in rat brain slices using antisera directed against the serotonin transporter. The density of serotonin transporter immunopositive fibers is greatest in the superior vestibular nucleus and the medial vestibular nucleus, especially along the border of the fourth ventricle; it declines in more lateral and caudal regions of the vestibular nuclear complex. After unilateral iontophoretic injections of Fluoro-Gold into the vestibular nuclei, retrogradely labeled neurons were found in the dorsal raphe nucleus (including the dorsomedial, ventromedial and lateral subdivisions) and nucleus raphe obscurus, and to a minor extent in nucleus raphe pallidus and nucleus raphe magnus. The combination of retrograde tracing with serotonin immunohistofluorescence in additional experiments revealed that the vestibular nuclei receive both serotonergic and non-serotonergic projections from raphe nuclei. Tracer injections in densely innervated regions (especially the medial and superior vestibular nuclei) were associated with the largest numbers of Fluoro-Gold-labeled cells. Differences were observed in the termination patterns of projections from the individual raphe nuclei. Thus, the dorsal raphe nucleus sends projections that terminate predominantly in the rostral and medial aspects of the vestibular nuclear complex, while nucleus raphe obscurus projects relatively uniformly throughout the vestibular nuclei. Based on the topographical organization of raphe input to the vestibular nuclei, it appears that dense projections from raphe nuclei are colocalized with terminal fields of flocculo nodular lobe and uvula Purkinje cells. It is hypothesized that raphe-vestibular connections are organized to selectively modulate processing in regions of the vestibular nuclear complex that receive input from specific cerebellar zones. This represents a potential mechanism whereby motor activity and behavioral arousal could influence the activity of cerebellovestibular circuits. PMID- 12890527 TI - Quorex Pharmaceuticals, Inc. Taking the bull by the horns. PMID- 12890528 TI - New insights into drug resistance in cancer. AB - New findings by Kavallaris et al. characterize mechanisms of resistance to epothilones and paclitaxel in cell lines. One significant discovery is a novel tubulin point mutation positioned outside the paclitaxel binding site, which confers a high degree of drug resistance. PMID- 12890529 TI - RNA as a transcriptional activator. AB - Two recent reports demonstrate that in vivo selection can isolate novel RNAs that activate transcription when tethered to a gene promoter. This highlights the structural plasticity that allows RNA to fulfill many functions normally carried out by proteins. PMID- 12890530 TI - Death, taxes, and the genetic code? AB - The genetic code is an enduring feature of biology: only rare circumstances result in changes to translation of the code, at least in nature. Researchers are devising methods to engineer ribosome-synthesized polypeptides containing novel and potentially useful amino acids. PMID- 12890531 TI - Screening for inducers of kinase degradation. AB - Targeted small molecule-induced protein degradation is a promising approach to inhibit signaling within kinase cascades. In this issue, researchers describe a simple assay for the rapid, high-throughput identification of novel agents that promote degradation of the kinases Her2 and EGFR. PMID- 12890532 TI - Sequence-universal recognition of duplex DNA by oligonucleotides via pseudocomplementarity and helix invasion. AB - The well-known Watson-Crick complementarity rules, which were discovered 50 years ago, elegantly direct the specific pairing of two DNA single strands. On the contrary, once formed, the double-stranded (ds) DNA lacks such a simple and sequence-universal recognition principle, since most of the characteristic chemical groups of nucleobases are now buried deep inside the double helix, the major DNA form. We report a promising versatile approach for highly selective recognition of designated sites within dsDNA featuring considerable practical potential for a variety of molecular-biological, biotechnological, gene therapeutic, and diagnostic applications. It may also have implications for prebiotic evolution of genetic machinery at the primordial stages of the origin of life. Our design synergistically employs the robust helix-invasion ability of recently developed DNA mimics and analogs, pseudocomplementary peptide nucleic acids and pseudocomplementary oligonucleotides, thus enabling the sequence unrestricted recognition of chosen DNA duplexes by nucleobase oligomers. Using this basically general approach, we selectively tagged a unique mixed-base site on the target dsDNA fragment with streptavidin and/or multiply labeled this site with fluorophores via the primer-extension reaction. PMID- 12890533 TI - Microtubule alterations and mutations induced by desoxyepothilone B: implications for drug-target interactions. AB - Epothilones, like paclitaxel, bind to beta-tubulin and stabilize microtubules. We selected a series of four leukemia sublines that display increasing levels of resistance to the epothilone analog desoxyepothilone B (dEpoB). The dEpoB cells selected in 30-140 nM were approximately 15-fold cross-resistant to paclitaxel, while 300 nM selected cells were 467-fold resistant to this agent. The dEpoB selected cells are hypersensitive to microtubule destabilizing agents, and express increased levels of class III beta-tubulin and MAP4. A novel class I beta tubulin mutation, A231T, that affects microtubule stability but does not alter paclitaxel binding, was identified. The 300 nM selected cells acquired a second mutation, Q292E, situated near the M loop of class I beta-tubulin. These cells fail to undergo drug-induced tubulin polymerization due to dramatically reduced drug binding. The dEpoB-resistant leukemia cells provide novel insights into microtubule dynamics and, in particular, drug-target interactions. PMID- 12890534 TI - Mitochondrial-targeted fatty acid analog induces apoptosis with selective loss of mitochondrial glutathione in promyelocytic leukemia cells. AB - Some fatty acids and derivatives are known to induce cell death in cancer cells. Mitochondria may have important roles in the death process. Therefore, we investigated the mitochondrial contribution in cell death induced by a modified fatty acid, tetradecylthioacetic acid (TTA), which cannot be beta-oxidized. TTA treatment induced apoptosis in IPC-81 leukemia cells via depolarization of the mitochondrial membrane potential (deltapsi) and early release of cytochrome c, accompanied by depletion of mitochondrial glutathione. Caspase-3 activation and cleavage of poly (ADP-ribose) polymerase (PARP) occurred at a late stage, but the broad-spectra caspase inhibitor zVAD-fmk did not block TTA-induced apoptosis. Overexpression of Bcl-2 partially prevented TTA-induced apoptosis, whereas cAMP induced cell death was completely blocked. In conclusion, TTA seems to trigger apoptosis through mitochondrial-mediated mechanisms and selective modulation of the mitochondrial redox equilibrium. PMID- 12890535 TI - Anatomy of glycosynthesis: structure and kinetics of the Humicola insolens Cel7B E197A and E197S glycosynthase mutants. AB - The formation of glycoconjugates and oligosaccharides remains one of the most challenging chemical syntheses. Chemo-enzymatic routes using retaining glycosidases have been successfully harnessed but require tight kinetic or thermodynamic control. "Glycosynthases," specifically engineered glycosidases that catalyze the formation of glycosidic bonds from glycosyl donor and acceptor alcohol, are an emerging range of synthetic tools in which catalytic nucleophile mutants are harnessed together with glycosyl fluoride donors to generate powerful and versatile catalysts. Here we present the structural and kinetic dissection of the Humicola insolens Cel7B glycosynthases in which the nucleophile of the wild type enzyme is mutated to alanine and serine (E197A and E197S). 3-D structures reveal the acceptor and donor subsites and the basis for substrate inhibition. Kinetic analysis shows that the E197S mutant is considerably more active than the corresponding alanine mutant due to a 40-fold increase in k(cat). PMID- 12890536 TI - Microtiter cell-based assay for detection of agents that alter cellular levels of Her2 and EGFR. AB - Overexpression of the transmembrane tyrosine kinases Her2 and EGFR is associated with aggressive malignancies, and several therapeutic strategies targeting the two receptors are now in various stages of clinical development. Most of the known agents block the activation or inhibit the activity of the kinases; however, a more significant therapeutic outcome may result from degrading these oncoproteins. Here, we report the development of a microtiter cell-based assay that sensitively detects cellular levels of Her2 and EGFR. The assay is useful in identifying small molecules that alter cellular levels of these kinases and in quantifying their effect. The method gives results comparable to Western blot, but it is faster, less labor intensive, and amenable to high throughput. PMID- 12890537 TI - DNA binding hairpin polyamides with antifungal activity. AB - Eight-ring hairpin polyamides containing N-methylimidazole (Im) and N methylpyrrole (Py) amino acids have been shown to bind with subnanomolar affinity to discrete DNA sites and to modulate a variety of DNA-dependent biological processes. We show here that addition of a second positive charge at the C terminus of an 8-ring hairpin polyamide confers activity against a number of clinically relevant fungal strains in vitro, and activity against Candida albicans in a mouse model. Control experiments indicate that the observed antifungal activity results from a DNA binding mechanism-of-action that does not involve DNA damage or disruption of chromosomal integrity. Hairpin activity is shown to be proportional to yeast DNA content (ploidy). Transcriptional interference is proposed as the likely explanation for fungal cytotoxicity. Experiments with sensitized yeast strains indicate the potential for discrete sites of action rather than global effects. PMID- 12890538 TI - RNA LEGO: magnesium-dependent formation of specific RNA assemblies through kissing interactions. AB - The high affinity and specificity of nucleic acid base complementarity has been proven to be a powerful method for constructing specific molecular assemblies. On the other hand, recent structural studies of RNA have revealed the wide range of tertiary interactions utilized in RNA folding, which may potentially be used as tools for the design of specific macromolecular assemblies. Here, RNA building blocks containing two hairpin loops, based on the dimerization initiation site (DIS) of HIV RNA, connected by a short linker were used to construct large RNA assemblies through hairpin loop-loop ("kissing") interactions. We show that specific linear and circular assemblies can be constructed in a magnesium dependent manner using several non-self-complementary loop-loop interactions designed in this study. These results show that the use of RNA tertiary interactions may broaden the repertoire of nucleic acid-based nanostructures. PMID- 12890539 TI - A versatile tRNA aminoacylation catalyst based on RNA. AB - Aminoacyl-tRNA synthetase (ARS) ribozymes have potential to develop a novel genetic coding system. Although we have previously isolated such a ribozyme that recognizes aromatic amino acids, it could not be used as a versatile catalyst due to its limited ability of aminoacylation to a particular tRNA used for the selection. To overcome this limitation, we used a combination of evolutionary and engineering approaches to generate an optimized ribozyme. The ribozyme, consisting of 45 nucleotides, displays a broad spectrum of activity toward various tRNAs. Most significantly, this ribozyme is able to exhibit multiple turnover activity and charge parasubstituted Phe analogs onto an engineered suppressor tRNA (tRNA(Asn)(CCCG)). Thus, it provides a useful and flexible tool for the custom synthesis of mischarged tRNAs with natural and nonnatural amino acids. PMID- 12890541 TI - Glycolysis and pyruvate oxidation in cardiac hypertrophy--why so unbalanced? AB - Cardiac hypertrophy, induced by chronic pressure or volume overload, is associated with abnormalities in energy metabolism as well as characteristic increases in muscle mass and alterations in the structure of the heart. Hypertrophied hearts display increased rates of glycolysis and overall glucose utilization, but rates of pyruvate oxidation do not rise in step with rates of pyruvate generation. Glycolysis and glucose oxidation, therefore, become markedly less 'coupled' in hypertrophied hearts than in non-hypertrophied hearts. Because the pyruvate dehydrogenase complex (PDC) contributes so powerfully to the control of glucose oxidation, we set out to test the hypothesis that the function of PDC is impaired in cardiac hypertrophy. In this review we describe evidence indicating that the alterations in glucose metabolism in hypertrophied hearts cannot be explained simply by changes in PDC expression or control. Additional mechanisms that may lead to an altered balance of pyruvate metabolism in cardiac hypertrophy are discussed, with commentaries on possible changes in pyruvate transport, NADH shuttles, lactate dehydrogenase, and amino acid metabolism. PMID- 12890540 TI - Preferred RNA binding sites for a threading intercalator revealed by in vitro evolution. AB - In pursuit of small molecules capable of controlling the function of RNA targets, we have explored the RNA binding properties of peptide-acridine conjugates (PACs). In vitro evolution (SELEX) was used to isolate RNAs capable of binding the PAC Ser-Val-Acr-Arg, where Acr is an acridine amino acid. The PAC binds RNA aptamers selectively and with a high degree of discrimination over DNA. PAC binding sites contain the base-paired 5'-CpG-3' sequence, a known acridine intercalation site. However, RNA structure flanking this sequence causes binding affinities to vary over 30-fold. The preferred site (K(D) = 20 nM) contains a base-paired 5'-CpG-3' step flanked on the 5' side by a 4 nt internal loop and the 3' side by a bulged U. Several viral 5'- and 3'-UTR RNA sequences that likely form binding sites for this PAC are identified. PMID- 12890542 TI - Sub-lethal plasma ammonia accumulation and the exercise performance of salmonids. AB - The proposal that plasma ammonia accumulation might impair the swimming performance of fish was first made over a decade ago, and has now proven to be the case for a number of salmonid species. The first experimental evidence was indirect, when a negative linear relationship between plasma ammonia concentrations and maximum sustainable swimming speed (U(crit)) was found following the exposure of brown trout (Salmo trutta) to sub-lethal concentrations of copper in soft acidic water. Since then, negative linear relationships between plasma ammonia concentration and U(crit) have been demonstrated following exposure of brown trout, rainbow trout (Oncorhynchus mykiss) and coho salmon (Oncorhynchus kisutch) to elevated water ammonia. For brown trout, the relationships between plasma ammonia and U(crit) were remarkably similar following either exposure to elevated water ammonia or to sub-lethal copper. This indicates that the impairment of swimming performance resulting from exposure to sub-lethal concentrations of heavy metals may be attributable in large part to an accumulation of endogenous ammonia. The negative relationship between plasma ammonia concentration and U(crit) was similar in size-matched rainbow and brown trout but, under similar regimes of ammonia exposure, rainbow trout were able to maintain a significantly lower plasma ammonia concentration, revealing inter specific differences in ammonia permeability and/or transport. One primary mechanism by which ammonia accumulation may impair exercise performance is a partial depolarisation of membrane potential in tissues such as the brain and white muscle. This may prejudice the co-ordination of swimming movements and reduce or abolish the development of muscle tension, thus, compromising swimming efficiency and performance at the top end of the range. PMID- 12890543 TI - The effect of copper on osmoregulation in the freshwater amphipod Gammarus pulex. AB - The influence of copper on osmoregulation in the freshwater amphipod Gammarus pulex was determined from the analysis of water permeability, haemolymph sodium concentration, sodium influx and gill Na(+)/K(+) ATPase and Mg(2+) ATPase activity. Exposure to nominal copper concentrations of 100 microg l(-1) or greater caused a significant reduction in both haemolymph sodium concentration and sodium influx within 4 h. Measurements of water permeability, expressed as the half-time of exchange of body water (t(1/2)), excluded structural gill damage as the cause of this fall in haemolymph sodium. Copper at 10 microg l(-1) or above in the assay solution significantly reduced gill Na(+)/K(+) ATPase activity. In contrast gill Mg(2+) ATPase activity was markedly less affected by copper. These differences in enzyme sensitivity were considered with respect to the potential mechanisms of copper toxicity. PMID- 12890544 TI - Oxidative stress in streptozotocin-induced diabetic rats: effects of garlic oil and melatonin. AB - In the present study, oxidative stress in diabetic model and the effect of garlic oil or melatonin treatment were examined. Streptozotocin (60 mg/kg body weight, i.p.)-induced diabetic rats, showed a significant increase of plasma glucose, total lipids, triglyceride, cholesterol, lipid peroxides, nitric oxide and uric acid. Concomitantly, significant decreases in the levels of antioxidants ceruloplasmin, albumin and total thiols were found in the plasma of diabetic rats. Lipid peroxide levels were significantly increased in erythrocyte lysate and in homogenates of liver and kidney, while superoxide dismutase (SOD) activities were decreased in tissue homogenates of liver and kidney. Treatment of diabetic rats with garlic oil (10 mg/kg i.p.) or melatonin (200 microg/kg i.p.) for 15 days significantly increased plasma levels of total thiol, ceruloplasmin activities, albumin. Lipid peroxides, uric acid, blood glucose, total lipid, triglyceride and cholesterol were decreased significantly after treatment with garlic oil or melatonin. Nitric oxide levels were decreased significantly in rats treated with melatonin only. In erythrocytes lysate, glutathione S-transferase (GST) activities were increased significantly in rats treated with garlic oil or melatonin, while lipid peroxides decreased significantly and total thiol increased significantly in melatonin or garlic oil treatment, respectively. In liver homogenates of rats treated with garlic or melatonin, lipid peroxides were decreased significantly, and GST activities increased significantly, while SOD activities were increased significantly in liver and kidney after garlic or melatonin treatment. The results suggest that garlic oil or melatonin may effectively normalize the impaired antioxidants status in streptozotocin induced diabetes. The effects of these antioxidants of both agents may be useful in delaying the complicated effects of diabetes as retinopathy, nephropathy and neuropathy due to imbalance between free radicals and antioxidant systems. Moreover, melatonin may be more powerful free radical scavenger than garlic oil. PMID- 12890545 TI - Chemical characterization of the oligosaccharides in milk of high Arctic harbour seal (Phoca vitulina vitulina). AB - Carbohydrates were extracted from high Arctic harbour seal milk, Phoca vitulina vitulina (family Phocidae). Free neutral oligosaccharides were separated by gel filtration and preparative thin layer chromatography, while free sialyl oligosaccharides were separated by gel filtration and then purified by ion exchange chromatography, gel filtration and high performance liquid chromatography. Oligosaccharide structures were determined by 1H-NMR spectroscopy. The structures of the neutral oligosaccharides were as follows: lactose, 2'-fucosyllactose, lacto-N-neotetraose, lacto-N-neohexaose, monofucosyl lacto-N-neohexaose and difucosyl lacto-N-neohexaose. Thus, all of the neutral saccharides contained lactose or lacto-N-neotetraose or lacto-N-neohexaose as core units and/or non-reducing alpha(1-2) linked fucose. These oligosaccharides have also been found in hooded seal milk. The structures of the silalyl oligosaccharides were: monosialyl lacto-N-neohexaose, monosialyl monofucosyl lacto-N-neohexaose, monosialyl difucosyl lacto-N-neohexaose and disialyl lacto-N neohexaose. These oligosaccharides contained lacto-N-neohexaose as core units, and one or two alpha(2-6) linked Neu5Ac, and/or non-reducing alpha(1-2) linked Fuc. The Neu5Ac residues were found to be linked to GlcNAc or penultimate Gal residues. The acidic oligosaccharides are the first to have been characterized in the milk of any species of seal. PMID- 12890546 TI - Scrotal heating stimulates panting and reduces body temperature similarly in febrile and non-febrile rams (Ovis aries). AB - It is known that heating the ram scrotum stimulates heat loss resulting in a decrease in body temperature and that during fever core temperature increases, but local scrotal thermoeffectors operate to maintain normal scrotal temperature. We have investigated whether scrotal warming influences core body temperature and the panting effector during fever generation. We measured rectal temperature, intrascrotal temperature, scrotal skin temperature and respiratory frequency in four adult Merino rams following intravascular injection of saline or lipopolysaccharide at an ambient temperature of 18-20 degrees C while scrotal skin temperature was maintained at 33 degrees C or elevated to 41 degrees C. Compared to maintaining normal scrotal temperature, heating the scrotum increased respiratory frequency and reduced rectal temperature by a similar amount following LPS as following saline. Fever was associated with decreased respiratory frequency compared to saline at both 33 and 41 degrees C scrotal temperature, suggesting that the fever was generated mainly by decreasing respiratory heat loss. We conclude that scrotal thermal afferent stimulation resulted in an offset for the set-point of body temperature regulation in both normothermic and febrile rams. PMID- 12890547 TI - Effects of hypotonic shock on intracellular pH in bovine articular chondrocytes. AB - Chondrocytes inhabit an unusual environment, in which they are repeatedly subjected to osmotic challenges as fluid is expressed from the extracellular matrix during static joint loading. In the present study, the effects of hypotonic shock on intracellular pH, pH(i), have been studied in isolated bovine articular chondrocytes using the pH-sensitive fluroprobe BCECF. Cells subjected to a 50% dilution rapidly alkalinised, by approximately 0.2 pH units, a sustained plateau being achieved within 300 s. The effect was not altered by inhibitors of pH regulators, such as amiloride, bafilomycin and SITS, but was absent when cells were subjected to hypotonic shocks in solutions in which Na(+) ions were replaced by NMDG(+). The response was found to be sensitive to Gd(3+) ions, blockers of stretch-activated cation channels. Alkalinisation was also inhibited by treatment with Zn(2+) ions, at a concentration reported to block voltage-activated H(+) channels (VAHC). Depolarisation using high K(+) solutions supplemented with valinomycin also induced intracellular alkalinisation. Measurements using a membrane potential (E(m)) fluorescent dye showed that E(m) was approximately -44 mV, but was depolarised by over 50 mV following HTS. The depolarisation was also inhibited by Na(+) substitution with NMDG(+) or treatment with Gd(3+). We conclude that in response to HTS the opening of a stretch-activated cation channel leads to Na(+) influx, which results in a membrane depolarisation. Subsequent activation of VAHC permits H(+) ion efflux along the prevailing electrochemcial gradient, leading to the alkalinisation, which we record. PMID- 12890548 TI - Developmental and diel profiles of plasma corticosteroids in the bullfrog, Rana catesbeiana. AB - Corticosteroids synergize with the thyroid hormone (TH) at late metamorphic stages and might have a role in the hormonal regulation of amphibian metamorphosis. This role could be influenced by diel fluctuations, particularly if the peak of the plasma corticoids changed in relation to the TH peaks. Diel variation in plasma corticosteroids was studied in Rana catesbeiana prometamorphic and climax tadpoles on 18:6, 12:12 and 6:18 light:dark (LD) cycles. Cortisol (hydrocortisone; HC) and aldosterone (ALDO) exhibited different, but LD cycle-specific, circadian fluctuations at prometamorphosis, whereas corticosterone (CORT) was undetectable (less than 1.18 ng/ml). HC, ALDO and CORT rhythms became synchronous at early metamorphic climax on all LD cycles, although the cosinor-derived acrophases, which occurred around the time of the dark:light transition, shifted approximately 6 h earlier from 18L:6D to 6L:18D. On both 18L:6D and 12L:12D, the acrophase of HC changed little from prometamorphosis to climax, whereas that of ALDO underwent a major phase shift. On 6L:18D, both the ALDO and the HC acrophases shifted at climax. These LD cycle-specific phase shifts of the diel rhythms placed the acrophases of the corticoids in different phase relationships to that of the previously determined thyroxine (T(4)) acrophase at climax, and may partially explain the influence of the light regimen on metamorphic timing. The pronounced diel variations in the corticoid concentrations from the troughs to the peaks show that hormone levels are a function of the time of day and the environmental lighting regimen, which need to be taken into account in measuring the level of plasma hormones in amphibians. The 24-h means calculated from the data of all the sampling times showed that only plasma ALDO and CORT, but not HC, rose markedly at climax, although there were significant LD cycle-related differences in the mean levels of both HC and ALDO at prometamorphosis, and in HC at climax. Additional work sampling at mid light showed that plasma CORT peaked at Stage XXIII, decreased at the end of climax, and remained low in the postmetamorphic froglet at 2.1 ng/ml. In the adult bullfrog, CORT was clearly the predominant corticosteroid at 34.3 ng/ml, whereas HC and ALDO levels were only approximately 1.3 ng/ml. PMID- 12890549 TI - Effects of experimental anemia on blood ion and acid-base status of turtles during submergence in aerated water at 3 degrees C. AB - The importance of blood hemoglobin to aquatic oxygen uptake by turtles (Chrysemys picta bellii) submerged in aerated water at 3 degrees C was tested by comparing the responses of anemic turtles (hematocrit approximately 6%) to turtles with normal hematocrits (hematocrit approximately 33%). All turtles were submerged for 42 days and blood samples were collected at 0, 7, 21, 32 and 42 days. Blood was analyzed for pH, PCO(2), PO(2), hematocrit, hemoglobin concentration ([Hb]) and plasma was analyzed for concentrations of lactate, glucose, Na(+), K(+), Ca(2+) and Mg(2+). Plasma [HCO(3)(-)] was calculated. [Hb] correlated closely with hematocrit levels. [Lactate] reached higher final values in anemic turtles (34.5+/-5.3 mmol l(-1)) than in normal turtles (14.5+/-4.6 mmol l(-1)) indicating a greater reliance of the anemic animals on anaerobic metabolism. Both groups compensated for acidosis by reduced PCO(2) and anemic turtles also had increased [Ca(2+)] and [Mg(2+)]. Blood pH fell significantly in the anemic turtles but not in the controls. Although the data indicate that the anemic turtles relied more on anaerobic metabolism than the controls, the effect was much less than expected on the basis of the reduced blood O(2) carrying capacity. Possible compensatory mechanisms utilized by the anemic turtles to minimize anaerobic metabolism are discussed. PMID- 12890550 TI - Swimming of pregnant rats at different water temperatures. AB - We studied the chronic effect of exercise during water immersion, associated with thermal stress (water temperature at 22, 35 and 40 degrees C) at an intensity of 80% of maximal work load supported in pregnant rats (P) and non-pregnant female rats (NP). P and NP were subdivided into three subgroups according to water temperature during exercise (P22 and NP22; P35 and NP35; P40 and NP40). The animals were submitted to daily swimming sessions of 10-15 min, for 19 days of pregnancy (P) or experimental conditions (NP). Plasma concentration of triglycerides, cholesterol, glucose, total protein, albumin and corticosterone were determined 24 h after the last exercise session. Weight gain and rectal temperature pre- and post-swimming session were also determined. The offspring were examined just after caesarian section on the 20th day of pregnancy to check weight, length and litter size. Pregnant rats showed an increase of triglycerides, reduction of glycemia, total protein and albumin and cholesterol (at 35 degrees C) when compared to non-pregnant animals. Such effects probably lead to an adequate delivery of substrate to the fetus and prepare the mother for lactation. Daily thermal stress did not modify metabolic responses to exercise in pregnant rats. Results also show a deleterious effect on offspring when the mother is exposed daily to extreme temperatures during swimming. These results suggest that water temperature (cold and hot) in swimming have to be considered to avoid damage in fetal development. PMID- 12890551 TI - Recombinant goldfish growth hormones (gfGH-I and -II) expressed in Escherichia coli have similar biological activities. AB - Complementary DNA regions coding for two different mature goldfish growth hormones (gfGH-I and gfGH-II) with four and five cysteine residues were cloned into the bacterial expression vector, pRSETA. The recombinant gfGH-I (five cysteines) and -II (four cysteines) were produced in Escherichia coli as the fusion proteins carrying N-terminal 6XHis tag, which facilitates purification by using metal chelating affinity chromatography under denaturing condition with urea. The recombinant hormones were further refolded by gradually removing the urea. Native gfGH was also purified from goldfish pituitary glands and served as a positive control in the present study. The native and recombinant hormones were tested in goldfish hepatic radioreceptor assay and in vitro Spi 2.1 promoter activation assay. Our results showed that the two recombinant gfGHs are biologically active, and they have similar biological activities despite their having different cysteine contents. PMID- 12890552 TI - Comparative water relations of adult and juvenile tortoise beetles: differences among sympatric species. AB - Relative abundance of two sympatric tortoise beetles varies between drought and 'wet' years. Differing abilities to conserve water may influence beetle survival in changing environments. Cuticular permeability (CP), percentage of total body water (%TBW), rate of water loss and percentage of body lipid content were determined for five juvenile stages and female and male adults of two sympatric species of chrysomelid beetles, the golden tortoise beetle, Charidotella bicolor (F.) and the mottled tortoise beetle, Deloyala guttata (Olivier). There were significant differences in %TBW and lipid content among juvenile stages. Second instars had the greatest difference in CP (37.98 and 11.13 microgcm(-2)h(-1)mmHg( 1) for golden and mottled tortoise beetles, respectively). Mottled tortoise beetles had lower CP and greater %TBW compared with golden tortoise beetles, suggesting that they can conserve a greater amount of water and may tolerate drier environmental conditions. This study suggests that juvenile response to environmental water stress may differentially affect the survival of early instars and thus affect the relative abundance of adult beetles in the field. This is supported by the low relative abundance of golden tortoise beetle larvae in a drought year and the higher abundance in two 'wet' years. PMID- 12890553 TI - The effect of an acute phase response on tissue carotenoid levels of growing chickens (Gallus gallus domesticus). AB - Plasma, liver and skin carotenoids decrease following infectious disease challenges. Since these challenges often involve substantial host pathology and chronic immune responses, the mechanism underlying altered carotenoid deposition is unclear. Therefore, changes in tissue carotenoid levels were examined during an acute phase response induced by lipopolysaccharide (LPS) or interleukin-1 (IL 1). In two experiments, chicks were hatched from carotenoid-deplete eggs (n=28, n=64, respectively) and fed 0, 8 or 38 mg carotenoids (lutein+canthaxanthin)/kg diet. For chicks fed 38 mg carotenoids, but not those fed 0 or 8 mg, LPS generally reduced plasma lutein, canthaxanthin and total carotenoids (P<0.05), and liver lutein, zeaxanthin, canthaxanthin and total carotenoids (P<0.05). Additionally, LPS reduced thymic total carotenoids (P=0.05) and increased thymocyte lutein (P=0.07), zeaxanthin (P=0.07) and total carotenoids (P=0.07). Finally, LPS increased bursal canthaxanthin (P<0.01), but had no effect on shank carotenoids (P>0.5). In chicks hatched from carotenoid-replete eggs (n=36) and fed dietary lutein (38 mg/kg diet), LPS reduced plasma and liver zeaxanthin and liver total carotenoids (P<0.05); IL-1 reduced plasma and liver lutein, zeaxanthin and total carotenoids (P<0.05). Therefore, an acute phase response plays a role in reduced tissue carotenoids during infectious disease. PMID- 12890554 TI - Identification of a second member of the ponticulin gene family and its differential expression pattern. AB - We have identified a homologue (ponB) of the ponticulin gene (ponA), an F-actin binding protein, in the expressed sequence tag library generated to mRNA isolated from fusion-competent cells of Dictyostelium discoideum. PonB is predicted to have many of the same characteristics as ponticulin. Both proteins are predicted to possess a cleaved signal peptide, a glycosyl anchor, an amphipathic beta strand structure and six conserved cysteines. Because of the sequence similarity and predicted conserved structures, this gene constitutes the second member of a ponticulin gene family. Unlike ponticulin, ponB is not expressed in axenically grown cells or during the asexual reproductive phase of D. discoideum. PonB is expressed by cells grown on bacterial lawns and by cells induced to be fusion competent, i.e., gametes. The expression of ponB correlates with the appearance of a new F-actin binding activity in cell lysates of bacterially grown ponA(-) cells. By immunofluorescence microscopy, ponB appears to be localized to vesicles and to the plasma membrane of bacterially grown cells. Because ponticulin is the major high-affinity link between the plasma membrane and the cytoskeleton, the ponticulin gene family is likely to be part of the redundant system of proteins involved in connecting the cytoskeleton to the plasma membrane. PMID- 12890555 TI - The human kallikrein protein 5 (hK5) is enzymatically active, glycosylated and forms complexes with two protease inhibitors in ovarian cancer fluids. AB - The kallikrein family is a group of 15 serine protease genes clustered on chromosome 19q13.4. Binding of kallikreins to protease inhibitors is an important mechanism for regulating their enzymatic activity and may have potential clinical applications. Human kallikrein gene 5 (KLK5) is a member of this family and encodes for a secreted serine protease (hK5). This kallikrein was shown to be differentially expressed at the mRNA and protein levels in diverse malignancies. Our objective was to study the enzymatic activity and the interaction of recombinant hK5 protein with protease inhibitors. Recombinant hK5 protein was produced in yeast and mammalian expression systems and purified by chromatography. HPLC fractionation, followed by ELISA-type assays, immunoblotting and radiolabeling experiments were performed to detect the possible interactions between hK5 and proteinase inhibitors in serum. Enzymatic deglycosylation was performed to examine the glycosylation pattern of the protein. The enzymatic activity of hK5 was tested using trypsin and chymotrypsin-specific synthetic fluorogenic substrates. In serum and ascites fluid, in addition to the free ( approximately 40 kDa) form, hK5 forms complexes with alpha(1)-antitrypsin and alpha(2)-macroglobulin. These complexes were detected by hybrid ELISA-type assays using hK5-specific coating antibodies and inhibitor detection antibodies. The ability of hK5 to bind to these inhibitors was further verified in vitro. Spiking of serum samples with 125I-labeled hK5 results in the distribution of the protein in two higher molecular mass (bound) forms, in addition to the unbound form. The hK5 mature enzyme is active and shows trypsin, but not chymotrypsin-like, activity. The pro-form of hK5 is not active. Recombinant hK5 shows a higher than predicted molecular mass due to glycosylation. hK5 is partially complexed with alpha(1)-antitrypsin and alpha(2)-macroglobulin in serum and ascites fluid of ovarian cancer patients. The recombinant protein is glycosylated and its mature form shows trypsin-like activity. PMID- 12890556 TI - Structural and functional characterization of two human V-ATPase subunit gene promoters. AB - The vacuolar-ATPase (V-ATPase) is a multi-subunit enzyme that couples ATP hydrolysis to proton pumping across membranes. V-ATPase genes are considered to be housekeeping genes and are expressed in human neoplastic tissue and in cell lines. We have isolated and characterized several genomic clones containing the 5'-end of the human V-ATPase genes. DNA sequence analysis of the promoters of two V-ATPase subunit genes, encoding C (ATP6C) and c (ATP6F), reveals GC-rich regions in the region of the first exon. Neither TATA- nor CCAAT-boxes were found in these promoters, but both GC-boxes and E-boxes were identified. Transient transfection analysis, using a series of 5' nested deletions of promoter luciferase constructs in human cancer cells, demonstrated that a positive cis acting regulatory region was present in these TATA-less promoters. The regions between -79 and -40 of the ATP6C promoter and between -245 and -99 of the ATP6F promoter were identified as being likely to be extremely important for basal promoter activity. Electrophoretic mobility shift assays (EMSA) of these cis regulatory regions revealed the basal promoter to be highly complex, with cooperative binding of several transcription factors, including Sp family members. These data identify the critical regulatory regions for both the ATP6C and ATP6F basal promoters and stress the functional importance of multiple protein complexes, involving the Sp family of transcription factors, in regulating gene expression. PMID- 12890557 TI - The chromosome 21 transcription factor ETS2 transactivates the beta-APP promoter: implications for Down syndrome. AB - The gene that codes for beta-amyloid precursor protein (beta-APP), a protein centrally involved in senile plaque formation in Down syndrome (DS) and Alzheimer's disease (AD), is located on chromosome 21. In DS beta-APP expression is three- to fourfold higher than what is expected from the 1.5-fold increased gene load, suggesting that other genes on chromosome 21 directly or indirectly can further up-regulate beta-APP. Here we show that the chromosome 21 transcription factor ETS2 transactivates the beta-APP gene via specific Ets binding sites in the beta-APP promoter and, in this respect, cooperates with the transcription factor complex AP1. We further show that brains and primary neuronal cultures from Ets2 transgenic mice, as well as 3T3 fibroblasts that overexpress ETS2, display molecular abnormalities also seen in DS, such as elevated expression of beta-APP protein, an increase in presenilin-1 and increased beta-amyloid production. We conclude that ETS2 is a transcriptional regulator of beta-APP and that overexpression of ETS2 in DS may play a role in the pathogenesis of the brain abnormalities in DS and possibly AD. PMID- 12890558 TI - Characterization of ciprofloxacin binding to the linear single- and double stranded DNA. AB - The binding of ciprofloxacin to natural and synthetic polymeric DNAs was investigated at different solvent conditions using a combination of spectroscopic and hydrodynamic techniques. In 10 mM cacodylate buffer (pH 7.0) containing 108.6 mM Na(+), no sequence preferences in the interaction of ciprofloxacin with DNA was detected, while in 2 mM cacodylate buffer (pH 7.0) containing only 1.7 mM Na(+), a significant binding of ciprofloxacin to natural and synthetic linear double-stranded DNA was observed. At low ionic strength of solution, ciprofloxacin binding to DNA duplex containing alternating AT base pairs is accompanied by the largest enhancement in thermal stability (e.g. DeltaT(m) approximately 10 degrees C for poly[d(AT)].poly[d(AT)]), and the most pronounced red shift in the position of the maximum of the fluorescence emission spectrum (lambda(max)). Similar red shift in the position of lambda(max) is also observed for ciprofloxacin binding to dodecameric duplex containing five successive alternating AT base pairs in the row. On the other hand, ciprofloxacin binding to poly[d(GC)].poly[d(GC)], calf thymus DNA and dodecameric duplex containing a mixed sequence is accompanied by the largest fluorescence intensity quenching. Addition of NaCl does not completely displace ciprofloxacin bound to DNA, indicating the binding is not entirely electrostatic in origin. The intrinsic viscosity data suggest some degree of ciprofloxacin intercalation into duplex. PMID- 12890559 TI - Molecular cloning and characterisation of two genes encoding asparagine synthetase in barley (Hordeum vulgare L.). AB - Two different cDNA clones encoding asperagine synthetase (AS: EC 6.3.5.4.) were cloned from barley (Hordeum vulgare L. cv. Alexis). The corresponding genes were designated HvAS1 (GenBank no AF307145) and HvAS2 (GenBank no AY193714). Chromosomal mapping using wheat-barley addition lines revealed that the HvAS1 gene is located on the long arm of barley chromosome 5, while the HvAS2 gene maps to the short arm of chromosome 3. Both genes are expressed in barley leaves according to RT-PCR analysis but only the HvAS1 gene expression can be detected in roots. Northern blots show no expression of HvAS1 in plants grown under a normal 16 h light/8 h dark cycle but after 10 h of continuous darkness, transcript appears and mRNA accumulates over a 48-h period of dark treatment. In roots, low-level expression of HvAS1 could be detected and the expression level appears to be unaffected by light. A polyclonal antibody was raised against the HvAS1 protein and used in Western blot analysis. The AS protein accumulated during a 48-h period of dark treatment, following the increase in HvAS1 transcript. PMID- 12890560 TI - Smooth muscle gamma-actin promoter regulation by RhoA and serum response factor signaling. AB - Smooth muscle gamma-actin (SMGA) is both an early marker of smooth muscle cell differentiation, which demonstrates an expression pattern restricted to smooth muscle and the post meiotic spermatocyte. Serum response factor (SRF) DNA-binding is an important regulator of muscle differentiation, including SMGA expression during smooth muscle cell differentiation. RhoA, a low molecular weight GTPase protein, can regulate cardiac, skeletal, and smooth muscle differentiation through SRF-dependent mechanisms. This study's purpose was to examine RhoA expression during smooth muscle cell development, and determine if the SMGA promoter activity is sensitive to RhoA-mediated signaling through SRF. Additionally, the study identified the promoter regulation modifying SMGA expression by RhoA signaling. Western blot analysis of embryonic chick gizzard whole protein extracts during 5 to 14 days of development demonstrated a large induction of RhoA (10-fold) and beta1 integrin expression at day 8, which corresponds to the time SMGA expression and differentiation are occurring. Transient transfections in CV-1 fibroblast cells demonstrated that co overexpression of SRF and RhoA could induce a 40-fold induction of -176 bp SMGA promoter activity. Mutational analysis demonstrated that serum response element (SRE)-1, but not SRE2, was necessary for RhoA/SRF activation of the SMGA promoter. Deletion analysis revealed that although SRE1 was necessary for SMGA promoter activation by RhoA and SRF, it was not sufficient, implicating a possible obligatory role of additional promoter sequences in the response. Overexpression of a mutated SRF protein that was unable to bind DNA demonstrated that the 40-fold RhoA/SRF activation was largely dependent on SRF binding to the SMGA promoter. Thus, as the SMGA promoter appears to be a target of RhoA-mediated transcriptional regulation, the uncovering of these signaling mechanisms effecting SMGA promoter activity should provide a regulatory paradigm that can then be examined during the regulation of other smooth muscle genes. PMID- 12890561 TI - A novel Xenopus laevis SRY-related gene, xSox33. AB - The sex-determining region Y (SRY) gene and its related Sox genes encode transcriptional regulatory factors. In this study, we isolated and sequenced a novel Sox cDNA from African clawed frog (Xenopus laevis). The Sox gene was named xSox33. xSox33 was revealed to encode 244 amino acids. Reverse transcription polymerase chain reaction (RT-PCR) showed that xSox33 was expressed at very low levels in some frog tissues including lung, ovary, skeletal muscle, testis, brain and heart. Its embryonic expression was also studied by RT-PCR. After the mid blastula transition, the zygotic expression was initiated during gastrulation and the level was elevated as the embryogenesis proceeded. Electrophoretic mobility shift assay (EMSA) indicated that a recombinant xSox33 polypeptide was capable of binding to the nucleotide sequence AACAAT. PMID- 12890562 TI - Murine granulosa cell morphology and function are regulated by a synthetic Arg Gly-Asp matrix. AB - Extracellular matrix (ECM) proteins are established regulators of granulosa cell survival, morphology, and differentiation. In this study, the roles of ECM adhesion peptide density on murine granulosa cell adhesion, morphology, and steroid secretion were probed using synthetic matrices. The synthetic matrix was fabricated from the polysaccharide alginate, which does not inherently support cell adhesion but can be modified with controlled densities of adhesion peptides (10(-4) to 2 x 10(-1) ng/cm(2)). GRM02, a murine granulosa cell line, and primary murine granulosa cells were cultured on alginate matrices modified by coupling of synthetic peptide sequences containing the Arg-Gly-Asp motif common to ECM proteins. Cells cultured on these peptide-modified surfaces (0.02, 0.2 ng/cm(2)) attached and spread, with morphologies specific to the peptide identity and density. Additionally, progesterone and estradiol secretion was a function of peptide density, with up to threefold increases compared to controls. These results indicate that the density and identity of adhesion peptides regulate granulosa cell function. This system provides a mechanism to examine the granulosa cell-ECM interactions that occur during follicle maturation. PMID- 12890563 TI - Activation of the cAMP-dependent protein kinase signaling pathway by luteinizing hormone in trout theca layers. AB - In the fish ovary, LH is the main factor regulating the production of steroids during the periovulatory period and its effects are believed to be mediated, at least partially, through the cAMP-dependent protein kinase (PKA) signaling pathway. However, there is no direct evidence for the presence of PKA in the fish ovary nor on the regulation of its activity by fish LH. Here, we show the identification of regulatory (R) and catalytic (C) subunits of PKA in trout theca cells by immunoblotting. DEAE-cellulose chromatography of theca cell extracts indicated the presence of PKA type I and II and showed that trout theca cells display PKA-specific phosphotransferase and cAMP-binding activities. Salmon LH (sLH) stimulated PKA activity and increased the levels of immunoreactive RIIalpha, RIIbeta and C subunits in trout theca layers. These observations, coupled with the sLH-dependent decrease in the half-life of the C subunit, as shown by pulse-chase experiments, strongly suggest that sLH activates PKA in trout theca cells. Furthermore, our results suggest that ovarian PKA activity and its regulation by LH has been well conserved from fish to humans. PMID- 12890564 TI - Androgens stimulate coordinated lipogenic gene expression in normal target tissues in vivo. AB - In prostate cancer cell lines in culture androgens cause a marked and coordinated upregulation of the expression of several lipogenic genes. Here, using castrated male Wistar rats as an experimental paradigm, we investigated whether coordinated androgen stimulation of lipogenic gene expression represents a more general physiological regulation in non-cancerous androgen-responsive cells as well. In typical target tissues for androgen action such as the ventral prostate and the lacrimal gland, androgen deprivation resulted in a marked reduction in the mRNA and protein levels of genes involved in fatty acid (fatty acid synthase and acetyl-CoA-carboxylase) and cholesterol synthesis (HMG-CoA-reductase and farnesyl diphosphate synthase). Readministration of testosterone immediately following orchidectomy restored the expression of all four genes. Substitution of testosterone by the non-aromatizable androgen dihydrotestosterone gave rise to comparable changes in the mRNA and protein levels of the lipogenic genes under investigation, confirming the involvement of the androgen receptor in the observed effects. In support of the coordinate nature of this regulation, androgen-induced upregulation of lipogenic gene expression is accompanied by an increase in the nuclear content of SREBP, a key lipogenic transcription factor. Taken together, these findings provide evidence for a coordinate regulation of lipogenic gene expression not only in prostate cancer cell lines in culture but also in non-cancerous androgen-responsive tissues in vivo. PMID- 12890565 TI - Expression of the mouse homologue for the human GCDFP-15/PIP gene during pre- and early post-natal development. AB - The function of the mouse submaxillary gland/prolactin inducible protein (mSMGP/mPIP), the homologue of the human gross cystic disease fluid protein 15 (GCDFP-15)/prolactin inducible protein (hPIP) remains unknown. The human gene, normally expressed in apocrine glands of healthy individuals, is aberrantly expressed in human breast cancers where it is regulated by hormones including androgens, and in prostate cancers. We have previously reported that in the adult mouse and rat, gene expression is tissue-specific for the salivary and lacrimal glands, and is hormonally regulated. In this study, we examine the endogenous pattern of mouse SMGP/PIP (mSMGP/mPIP) gene expression in mid- and late embryonic, and in early postnatal development. Gene expression was analyzed by RT PCR followed by Southern blot analysis, and by in situ hybridization. Gene expression was detected in the submandibular gland as early as embryonic day 14 (E14), a period that coincides with the initiation of submandibular gland development in the embryo, suggesting that mSMGP/mPIP may have a functional role in the developing gland. Nearing the end of gestation, E18, mSMGP/mPIP transcripts were localized in the proacinar cells of the gland, and gene expression continued to be maintained following birth. In addition, during early postnatal development, mSMGP/mPIP gene expression was detected in the other two major salivary glands, the sublingual and parotid, as well as in the lacrimal gland and in reproductive tissues. In the prostate, gene expression was turned off by 10 weeks of age. The spatial and temporal pattern of the mSMGP/mPIP gene expression, in addition to our recent demonstration that mSMGP/mPIP is found in mouse saliva and can bind bacteria, suggest that this protein may have a protective role in the mouse. PMID- 12890566 TI - Orexin-A expression in human peripheral tissues. AB - Orexin-A is a neuropeptide present in the brain and is known to regulate feeding and sleeping. In this study, we examined the systemic distribution of orexin-A in human tissues. Immunoreactivity for orexin-A was detected in ganglion cells of the thoracic sympathetic trunk, myenteric plexuses and endocrine cells of the gastrointestinal tract, islet cells of the pancreas and syncytiotrophoblasts and decidual cells of the placenta. In the gastrointestinal tract, orexin-A immunoreactivity was detected in the myenteric plexuses from 26 gestational weeks to birth. In double immunostaining in the pancreas, a great majority of insulin positive cells was simultaneously positive for orexin-A. mRNA expression for prepro-orexin was also detected in the kidney, adrenal gland, pancreas, placenta, stomach, ileum, colon and colorectal epithelial cells. These results suggest the production of orexin-A in various human peripheral tissues and orexin-A may also play important roles in some peripheral organs. PMID- 12890567 TI - Four naturally occurring mutations in the human GnRH receptor affect ligand binding and receptor function. AB - In the present study, we performed functional analyses of four mutations in the human GnRH receptor (GnRHR) gene, identified in patients with idiopathic hypogonadotropic hypogonadism. These mutations result in amino acid substitutions in the extracellular N-terminal domain (Thr32Ile), second extracellular loop (Cys200Tyr), third intracellular loop (Leu266Arg) and sixth transmembrane helix (Cys279Tyr). Immunocytochemical analysis of cells transfected with HA-tagged GnRHR constructs revealed that all four mutant receptors were present on the cell surface. However, all four mutant receptors failed to exhibit measurable specific GnRH binding and, except for Thr32Ile, any significant inositol phosphate accumulation after GnRH stimulation. In addition, Leu266Arg and Cys279Tyr receptors were unable to stimulate gonadotropin subunit or GnRHR gene promoter activity in response to GnRH. Interestingly, the Cys200Tyr mutant was able to stimulate gonadotropin subunit and GnRHR promoter activity, albeit with a higher EC(50) and a markedly reduced maximal response compared to wild type receptor. The Thr32Ile mutant was also able to stimulate gonadotropin subunit and GnRHR promoters, but with a further significant increase in EC(50). Similarly, this mutant partially retained the ability to activate extracellular signal-regulated kinase 1 and stimulate CRE-luciferase activity with an identical shift in EC(50). Taken together, the studies suggest that the Thr32Ile mutation reduces hGnRHR function primarily by reducing ligand binding affinity, and the Cys200Tyr mutation reduces cell surface receptor expression. All four amino acid substitutions interfered with ligand binding, and affected signal transduction and stimulation of gonadotropin and GnRHR gene expression in response to GnRH. PMID- 12890568 TI - Compartment-specific phosphorylation of rat thyroid hormone receptor alpha1 regulates nuclear localization and retention. AB - The thyroid hormone receptor alpha1 (TRalpha1) is a transcription factor, which can activate or repress gene expression in response to thyroid hormone. In addition, some of its actions, including DNA binding and transcriptional activation, are thought to be regulated by phosphorylation. Results presented here, using Xenopus oocyte microinjection assays, demonstrate that a phosphorylated form of rat TRalpha1 is present in the nucleus, whereas unphosphorylated TRalpha1 remains cytoplasmic. Changes in the phosphorylation state of TRalpha1 occur rapidly and point to the possibility that phosphorylation occurs in the nucleus. Furthermore, increasing the overall phosphorylation state of the cell leads to enhanced nuclear retention of TRalpha1, suggesting that compartment-specific phosphorylation regulates nuclear localization of TRalpha1. Enhanced nuclear retention of TRalpha1 is not dependent on phosphorylation of serine 12, a well-characterized casein kinase II site, nor is phosphorylation of this site necessary for import of TRalpha1 into the Xenopus oocyte nucleus. Similarly, mutational analysis in mammalian cells shows that nuclear localization and partitioning of TRalpha1 to the nuclear matrix are independent of serine 12 phosphorylation. Taken together, these studies suggest that phosphorylation of one or more sites in TRalpha1, excluding serine 12, enhances nuclear retention and/or inhibits nuclear export but is not directly involved in nuclear import. PMID- 12890569 TI - Mouse calbindin-D(9k) gene expression in the uterus during late pregnancy and lactation. AB - Calbindin-D(9k) (CaBP-9k) is a cytosolic calcium binding protein mainly expressed in duodenum, placenta and uterus. In order to understand the expression pattern and regulation of uterine CaBP-9k gene, the expression of CaBP-9k mRNA and its regulation by estrogen (E2) and progesterone (P4) were investigated in the mouse uterus during late pregnancy (from day 12 to 18) and lactation. The expression levels of uterine CaBP-9k, estrogen receptor alpha (ERalpha) and progesterone receptor (PR) mRNAs were measured by Northern blot analysis. The expression levels of mouse uterine CaBP-9k mRNA gradually increased from pregnancy day 16 (P16), peaked at P18 (6.0-fold vs. P12) and declined at birth and during lactation. The expression levels of ERalpha and PR mRNAs indicated a similar fluctuation as CaBP-9k mRNA, suggesting the role of sex steroids/receptors in the regulation of CaBP-9k gene. To investigate effect of steroid hormone on CaBP-9k mRNA expression, three groups of animals were injected (s.c) with steroid hormone antagonists (RU486, tamoxifen, and ICI182780), respectively. RU486, a P4 antagonist, induced a significant decrease in CaBP-9k mRNA expression at 48 (3.2 fold) and 72 h (3.8-fold). However, tamoxifen and ICI182780, E2 antagonists, had no effect on CaBP-9k mRNA expression. Combined treatment with RU486 and ICI182780 did not further decrease the expression level of CaBP-9k mRNA when compared with RU486 treatment at 48 and 72 h. In addition, the treatment with RU40555, a glucocorticoid/progesterone antagonist, resulted in a decrease at 48 and 72 h following treatment. These results indicate that E2 is not likely involved in the regulation of CaBP-9k gene in the mouse uterus during late pregnancy and lactation. In conclusion, the present results suggest that P4, not E2 is a key regulator of CaBP-9k mRNA expression during late pregnancy and lactation. PMID- 12890570 TI - Differential G protein coupling preference of mammalian and nonmammalian gonadotropin-releasing hormone receptors. AB - Recently, we have identified three distinct types of gonadotropin-releasing hormone receptor (GnRHR) in the bullfrog (designated bfGnRHR-1, bfGnRHR-2, and bfGnRHR-3). In the present study, we compared G protein coupling preference of mammalian and nonmammalian GnRHRs. In a transient expression system, stimulation of either bfGnRHRs or rat GnRHR by GnRH significantly increased both inositol phosphates (IP) and cAMP productions, but ratios of IP to cAMP induction levels were quite different among the receptors, indicating differential G protein coupling preference. Using cAMP-dependent protein kinase A (PKA)-specific (CRE luc) or protein kinase C (PKC)-specific reporter (c-fos-luc) systems, we further examined G(s) and G(q/11) coupling preference of these GnRHRs. Since activities of CRE-luc and c-fos-luc were highly dependent on cell types, GnRH-induced CRE luc or c-fos-luc activity was normalized by forskolin-induced CRE-luc or 12-O tetradecanoylphenol-13-acetate (TPA)-induced c-fos-luc activity, respectively. This normalized result indicated that bfGnRHR-2 couples to G(s) more actively than G(q/11), while bfGnRHR-1 and -3 couple to G(s) and G(q/11) with similar strength. However, the rat GnRHR appeared to couple to G(q/11) more efficiently than G(s). This study was further confirmed by an experiment in which GnRH augmented CRE-driven luciferase activity in alphaT3-1 cells when CRE-luc was cotransfected with bfGnRHRs but not with vehicle or rat GnRHR. Collectively, these results indicate that mammalian and nonmammalian GnRHRs may induce diverse cellular and physiological responses through differential activation of PKA and PKC signaling pathways. PMID- 12890571 TI - Insulin-like growth factor-1 prevents loss of electrochemical gradient in cardiac muscle mitochondria via activation of PI 3 kinase/Akt pathway. AB - Insulin-like growth factor-1 (IGF 1) suppresses myocardial apoptosis and improves myocardial function in experimental models of cardiomyopathy. Apoptosis is triggered by mitochondria dysfunction and subsequent activation of caspases. We had previously shown that IGF 1 inhibited cardiomyocyte apoptosis via suppression of caspase, however, how IGF 1 and its signaling pathway modulates mitochondria function in cardiac muscle is not yet known. In this study we investigated how IGF 1 signaling modulates mitochondria membrane depolarization in the cardiomyocytes treated with doxorubicin. Doxorubicin rapidly induced loss of mitochondria electrochemical gradient and triggered mitochondria depolarization in primary cardiomyocytes, whereas addition of IGF 1 restored mitochondria electrochemical gradient. The effects of IGF 1 was blocked by a chemical inhibitor of PI 3 kinase and a dominant negative Akt, suggesting that IGF 1 signaling to mitochondria involves the PI 3 kinase-Akt pathway. Transducing cardiomyocytes with constitutive active PI 3 kinase partially restored the mitochondria electrochemical gradient in doxorubicin-treated cells. These findings provide direct evidence that IGF 1 modulation of mitochondria function is mediated through activation of PI 3 kinase and Akt. Additional experiments using agonist and antagonist of mitochondria K(ATP) channel suggest that IGF 1 signaling to mitochondria membrane does not directly involve K(ATP) channel. These findings suggest that cytosolic signaling to mitochondria may play a fundamental role in the cardiotoxic actions of doxorubicin and cardioprotective actions of IGF 1. PMID- 12890572 TI - Thyroid hormone upregulates ecto-5'-nucleotidase/CD73 in C6 rat glioma cells. AB - Thyroid hormones have profound effects on the central nervous system, such as proliferation, secretion of growth factors and gene expression regulation. Ecto NTPDases and ecto-5'-nucleotidase can control the extracellular ATP/adenosine levels, which have been described as proliferation factors. Here, we investigated the influence of T(3) on the enzyme cascade which catalyzes interconversion of purine nucleotides in rat C6 glioma cells. Exposure of C6 cells to T(3) caused a dose dependent increase of 30% in the AMP hydrolysis up to 0.25 nM, which was suppressed by actinomycin. No significant alteration was observed on ATP/ADP hydrolysis and T(4) at higher concentrations (10-1000 nM) promoted an increase in AMP hydrolysis that was not dose dependent. T(3) treatment also increased the expression of CD73 mRNA. Besides the importance of the ecto-5'-NT in the cell proliferation and differentiation, its overexpression can enhance extracellular adenosine levels, which could also be an important proliferation signal. PMID- 12890573 TI - Insulin attenuates leptin-induced STAT3 tyrosine-phosphorylation in a hepatoma cell line. AB - Leptin, the 16 kDa protein product of the ob gene, is secreted by adipocytes. The long form leptin receptor (ObRb) is expressed at high levels in the hypothalamus, and regulates appetite and energy expenditure. The fact that serum concentration of leptin is correlated with body mass index (BMI) suggests reduced sensitivity to leptin. Even though hyperinsulinemia and hyperleptinemia could coexist in obese humans, little is known about the interaction of insulin and leptin. In this study, we examined the effect of insulin on leptin signaling using Huh 7 cells transiently transfected with ObRb cDNA. Insulin inhibits leptin-induced STAT3 phosphorylation in a time- and dose-dependent manner without affecting Janus tyrosine kinases (JAKs) JAK2 phosphorylation. Okadaic acid prevents the inhibitory effect of insulin on leptin-induced STAT3 activation. PMID- 12890574 TI - Transcriptional regulation of the human Runx2/Cbfa1 gene promoter by bone morphogenetic protein-7. AB - It is well established that core binding factor Runx2/Cbfa1 is required for osteoblast recruitment and differentiation from mesenchymal stem cells. Transcriptional regulation of the Runx2/Cbfa1 gene by osteogenic factors such as bone morphogenetic proteins (BMPs) plays an important role in the stimulation of bone formation by these cytokines. BMP7 (also termed OP-1) is a member of the transforming growth factor beta (TGF-beta) superfamily and induces osteoblast differentiation from mesenchymal precursor stem cells in vitro as well as bone formation in vivo. This study examines the effects of BMP7 on markers of osteoblast differentiation and specifically on human Runx2/Cbfa1 gene transcription in a mouse C2C12 myoblast cell line where it induces expression of both alkaline phosphatase (ALP) and endogenous Runx2/Cbfa1. To further understand the mechanisms of human Runx2/Cbfa1 transcriptional regulation by BMP7, we cloned 3.0 kb of the human Runx2/Cbfa1 gene 5'-upstream flanking region and created a series of promoter deletions cloned into luciferase-based reporter vectors (Runx2/Cbfa1/Luc). Sequence data revealed six copies of the osteoblastic cis acting element (OSE2) in the proximal promoter region. In C2C12 cells transiently transfected with Runx2/Cbfa1/Luc deletion constructs, transcriptional activity of Runx2/Cbfa1 was upregulated up to 2-fold after 24 h of BMP7 treatment. Mutational analysis demonstrated that the minimal responsive promoter region for BMP7 regulated transcription maps to a proximal -74 OSE2 site. Electromobility shift assays with C2C12 cellular extracts indicate that BMP7 increases binding of OSE2 promoter sequences, and supershift assays with anti-Runx2/Cbfa1 antibodies demonstrate that Runx2/Cbfa1 is part of the nucleoprotein complex binding OSE2. Together, these data indicate BMP7 can upregulate Runx2/Cbfa1 gene expression in C2C12 myoblast cells, and suggest that Runx2/Cbfa1 may bind to OSE2 elements within its own promoter to autoregulate gene transcription in differentiating osteoblasts. PMID- 12890576 TI - Modulation of the Na(+)-H(+) antiport activity by adrenaline on erythrocytes from normal and obese individuals. AB - The effect of adrenaline on normal and obese human Na(+)-H(+) antiport (NHE 1) erythrocyte activity has been studied. Adrenaline increased both intracellular pH (pHi) and Na(+) influx in erythrocyte suspensions. This effect of adrenaline was inhibited by amiloride or EIPA, indicating that adrenaline stimulated NHE 1. Phorbol myristicate ester (PMA), a protein kinase C (PKC) stimulator, increased the activity of NHE 1 whereas calphostin C, a PKC inhibitor, partially inhibited NHE 1 activation induced by adrenaline. The effect of adrenaline to NHE 1 was counteracted by prazocin and by propranolol as well indicating the involvement of both alpha and beta 2 adrenergic receptors. The effect of adrenaline on erythrocyte NHE 1 activity was significantly more profound in obese compared to normal subjects. These data indicate that adrenaline induces an increase of pHi and Na(+) uptake of human erythrocytes through stimulation of NHE 1 activity. The significantly more profound stimulation of NHE 1 activity by adrenaline in obese as compared to normal subjects is discussed. PMID- 12890575 TI - Expression pattern and functional studies of matrix degrading proteases and their inhibitors in the mouse corpus luteum. AB - The formation of the corpus luteum (CL) is accompanied with angiogenesis and tissue remodeling and its regression involves tissue degradation. Matrix degrading proteases such as plasminogen activators (PAs) and matrix metalloproteinases (MMPs) are thought to play important roles in such controlled proteolytic processes. In this study, in situ hybridization has been used to examine the regulation and expression pattern of mRNAs coding for proteases and protease inhibitors belonging to the PA- and MMP-systems during the life cycle of the CL in an adult pseudopregnant mouse model. Of the nine proteases and five protease inhibitors that were studied, the majority were found to be temporally expressed during the formation and/or the regression of the CL. However, the mRNAs coding for urokinase type PA (uPA), membrane-type 1 MMP (MT1-MMP), and tissue inhibitor of metalloproteinases type-3 (TIMP-3) were constantly expressed in the mouse CL throughout its whole life span. To study the functional role of uPA in the CL, we analyzed luteal formation and function in uPA deficient mice. Our results revealed no significant difference in ovarian weight, serum progesterone levels, and blood vessel density in the functional CL between uPA deficient and wild type control mice. The temporal and spatial expression pattern of proteases and protease inhibitors during the CL life span suggests that members of the PA- and MMP-systems may play important roles in the angiogenesis and tissue remodeling processes during CL formation, as well as in the tissue degradation during luteal regression. However, the absence of reproductive phenotypes in mice lacking uPA and several other matrix degrading proteases indicates that there are redundancies among different matrix degrading proteases or that tissue remodeling in the ovary may involve other additional unique elements. PMID- 12890577 TI - Inhibition of IL-1beta-dependent prostaglandin E2 release by antisense microsomal prostaglandin E synthase 1 oligonucleotides in A549 cells. AB - The metabolism of arachidonic acid through the cyclooxygenase pathway is a highly regulated cellular process that results in the formation of PGH2. This unstable intermediate can be enzymatically metabolized to PGE2 by the actions of a microsomal 17 kDa PGE synthase (mPGES1). Treatment of A549 cells with IL-1beta for 24 h resulted in a twofold increase in mPGES1 mRNA, protein expression, and PGES specific activity. To understand the relationship between expression of mPGES1 and PGE2 formation, IL-1beta treated cells were incubated with increasing concentrations of antisense oligonucleotides (ASO) and their effects compared to cells treated with reverse sense oligonucleotides (RSO) designed against the ATG translation initiation codon of mPGES1. Incubation with ASO resulted in a 44% reduction in mRNA expression level as compared to RSO-treated cells. Microsomal preparations isolated from ASO- and RSO-treated cells were analyzed for their ability to convert PGH2 to PGE2 in the presence 2.5 mM reduced glutathione. An approximate 50% reduction (ASO: 1.8 nmol/min/mg, RSO: 3.7 nmol/min/mg) in PGES activity, protein expression by immunodetection, and extracellular PGE2 release was detected in these samples. As a control in these studies, the protein levels of COX2 and secreted IL-8 were quantified; no change in these levels was observed. These results demonstrate the direct association between mPGES1 expression, its enzymatic activity, and total PGE2 production following an inflammatory stimulus. PMID- 12890578 TI - Prothoracicotropic hormone stimulated extracellular signal-regulated kinase (ERK) activity: the changing roles of Ca(2+)- and cAMP-dependent mechanisms in the insect prothoracic glands during metamorphosis. AB - The synthesis of ecdysteroids by the lepidopteran prothoracic gland is regulated by a brain neuropeptide hormone, prothoracicotropic hormone (PTTH). In Manduca sexta glands, PTTH stimulates several events including Ca(2+) influx, Ca(2+) dependent cAMP generation and the activation of several protein kinases. In the present study, the path by which PTTH stimulates extracellular signal-activated regulated kinase (ERK) phosphorylation was investigated using PTTH and second messenger analogs. The results indicate that Ca(2+)-dependent processes, other than cAMP generation, play the major role in PTTH stimulation of ERK phosphorylation in larval prothoracic glands, that cAMP-dependent events increase in importance during later development and that PTTH-stimulated ERK phosphorylation is highest in larval glands. The decline in PTTH-stimulated ERK phosphorylation associated with metamorphosis results from decreased ERK levels and an increased basal rate of ERK phosphorylation. The data suggest that the role or importance of components of the PTTH signal transduction cascade are not fixed and can change during development. PMID- 12890579 TI - Peroxisome proliferator-activated receptor alpha induces rat sterol carrier protein x promoter activity through two peroxisome proliferator-response elements. AB - Sterol carrier protein x (SCPx) plays a critical role in the peroxisomal oxidation of fatty acids. It has been previously demonstrated in streptozotocin induced diabetic rats that SCPx expression is induced in association with an elevation in serum fatty acid and triglyceride levels. To elucidate the mechanisms underlying the expression of this gene during diabetes, the rat SCPx promoter was cloned and analyzed for regulatory motifs. Sequence analysis of this TATA-less promoter revealed two putative peroxisomal-proliferator-response element (PPRE) binding motifs at positions -134 and -869 relative to the translation start site. To examine peroxisomal-proliferator-activated receptor alpha (PPARalpha) effects on this gene, 935 bp of the SCPx promoter containing both PPRE motifs was cloned in front of the chloramphenicol acetyl-transferase gene or the luciferase gene and co-transfected into HTB-9 cells with vectors that encoded for PPARalpha and retinoid X receptor (RXR). The results indicate that PPARalpha was able to induce SCPx promoter activity in both cases, an effect that was enhanced by RXR and clofibrate. In addition, mutational analysis studies demonstrated that both PPREs contributed to the PPARalpha/RXRalpha-dependent activation of the SCPx promoter. Mobility shift assays and supershift analysis showed that nuclear extracts containing PPARalpha bound to the two PPRE motifs. This investigation indicates that similar to other genes involved in beta oxidation, SCPx transcription may be controlled by fatty acid levels via PPARalpha. PMID- 12890580 TI - Effect of thyroid state on H2O2 production by rat liver mitochondria. AB - It has been suggested that activation of mitochondrial respiration by thyroid hormone results in oxidative tissue injury secondary to increased reactive oxygen species production. In order to throw light on this subject, the effects of thyroid state on O2 consumption and H2O2 release by rat liver mitochondria were investigated. Hypothyroidism decreased the rates of O2 consumption and H2O2 release by succinate or pyruvate/malate-supplemented mitochondria during both State 4 and State 3 respiration, whereas hyperthyroidism increased such rates. Conversely, with both substrates and during either respiration phase, the percentage of O2 released as H2O2 was not significantly affected by thyroid state. On the other hand, the capacity of mitochondria to remove H2O2 increased by about 17% in hyperthyroid rats and decreased by about 35% in hypothyroid ones. This result indicates that the ratio between H2O2 production and release and so the percentage of O2 turned into H2O2 instead of being reduced to water increase in the transition from hypothyroid to hyperthyroid state. In light of previous observations that mitochondrial content of cytochromes and ubiquinone also increases in such a transition, the modifications of H2O2 production appear to be due to a modulation by thyroid hormone of the mitochondrial content of the autoxidisable electron carriers. This view is supported by measurements of H2O2 release in the presence of respiratory inhibitors, which show that the thyroid state-linked changes in H2O2 production occur at H2O2 generator sites of both Complex I and Complex III. PMID- 12890581 TI - Involvement of MAP kinase in the mediation of GH action on ovarian granulosa cells. AB - Growth hormone (GH), prostaglandins F (PGF) and prostaglandins E (PGE) are important regulators of ovarian function. Therefore, interrelationships between GH and these substances and their intracellular mechanisms might be of physiological significance in the ovary. The aims of this study on cultured porcine ovarian granulosa cells were to determine the effect of GH on the secretion of oxytocin (OT), PGF and PGE and whether MAP kinase could be involved in the mediation of GH action. Experiments were carried out with cultured porcine granulosa cells to investigate the effects of exogenous pGH (1-100 ng/ml) on the expression of MAP kinase (ERK-1, -2) and of PGH (1-100 ng/ml) and the MAP kinase blocker PD 98059 (1 microg/ml) on the secretion of PGF, PGE and OT. The cellular content of ERK-1 and -2 was analyzed by Western immunoblotting and immunocytochemistry, whilst PGF, PGE and OT accumulation in the medium was measured by RIA. Addition of GH to culture medium significantly altered the pattern of ovarian ERK MAP kinase on SDS-PA gels: the 44 and 42 kDa bands were reduced and additional 50 and 48 kDa bands appeared. Moreover, there was an increase in the percentage of cells containing ERK MAP kinase. GH stimulated the secretion of PGF (at a concentration of 1 ng GH per ml medium) and OT (100 ng GH per ml), but not PGE. The MAP kinase blocker alone did not affect PGF, PGE and OT secretion but did prevent the stimulatory effects of GH on PGF and induced stimulatory action of GH (10 ng/ml) on PGE. GH-stimulated OT secretion was unaffected. These observations confirm the role of GH in regulating porcine ovarian PGF, PGE and OT secretion and the presence of ERK MAP kinase in porcine granulosa cells. Furthermore, our studies demonstrate that MAP kinase-dependent intracellular mechanisms are dependent on GH, and that these mechanisms are involved in the mediation of GH action on ovarian PGF and PGE but not OT secretion. PMID- 12890582 TI - Deja vu: angiotensin and stress. PMID- 12890583 TI - The voltage sensor of ion channels revealed. PMID- 12890589 TI - Subclinical hypothyroidism: the case for treatment. PMID- 12890590 TI - Subclinical hypothyroidism: the case against treatment. PMID- 12890591 TI - Current concepts and novel applications of LH activity in ovarian stimulation. AB - Luteinizing hormone (LH) is a crucial physiological regulator of the human menstrual cycle. LH activity is also contained in many medications used to treat anovulation and to stimulate multiple folliculogenesis for assisted reproduction techniques. However, LH activity had previously been regarded as just a contaminant of follicle-stimulating hormone (FSH)-containing products and deemed potentially detrimental for reproductive function. Novel experimental and clinical evidence now suggests that the administration of pharmacological amounts of LH activity, instead of being harmful, is therapeutically advantageous, particularly in the support and modulation of ovarian folliculogenesis. The aim of this article is to provide an overview of the effects of LH activity administration in ovarian stimulation and to outline novel unconventional gonadotropin regimens that might improve the efficacy, safety and convenience of ovulation induction. PMID- 12890592 TI - The intrarenal renin-angiotensin system and diabetic nephropathy. AB - The renin-angiotensin system (RAS) is a coordinated cascade of proteins and peptide hormones, the principal effector of which is angiotensin II (ANG II). Evidence now indicates that the kidney regulates its function via a self contained RAS in a paracrine fashion. In diabetic nephropathy, the intrarenal generation of ANG II is increased, in spite of suppression of the systemic RAS. This increase can contribute to the progression of diabetic nephropathy via several hemodynamic, tubular and growth-promoting actions. ANG II induces insulin resistance. ANG II type-1 (AT(1)) and type-2 (AT(2)) receptors are downregulated in chronic diabetes, but decreased AT(2) receptor expression might contribute to early diabetic nephropathy by reducing AT(2) receptor-mediated beneficial actions that are counter-regulatory to those of the AT(1) receptor. AT(2) receptor stimulation might account for part of the renal protection seen with AT(1) receptor blockade. A rat model of accelerated diabetic nephropathy is the (mREN 2) 27 renin transgenic rat treated with streptozotocin in which both the intrarenal and extrarenal RAS is activated. PMID- 12890593 TI - Naturally occurring mutations of the extracellular Ca2+-sensing receptor: implications for its structure and function. AB - The extracellular Ca2+-sensing receptor is a member of the G-protein-coupled receptor family 3 in which agonists bind to a dimeric Venus-flytrap domain in the extracellular portion of the receptor. How agonist binding to this domain leads to activation of the seven-transmembrane domain is a major unresolved question. Information derived from the three-dimensional structure of the Venus-flytrap domain of the related metabotropic glutamate type 1 receptor, and from naturally occurring mutations of the Ca2+-sensing receptor identified in subjects with familial hypocalciuric hypercalcemia and autosomal dominant hypocalcemia offers new insights into the mechanism of receptor activation, and into the mechanism of action of allosteric modulators of the receptor. PMID- 12890594 TI - Sex hormones and male health: effects on components of the frailty syndrome. AB - Physicians are seeing an increasing number of older male patients with chronic diseases and conditions. However, the potential relevance of low levels of circulating endogenous androgens in connection with these diseases and conditions is generally poorly understood. Research findings have suggested that androgens play a distinct role in bone metabolism, body composition such as muscle and fat mass and fat distribution, cognitive functioning, mood and well being. The aim of this paper is to summarize the currently available data on the association between endogenous androgens and the intermediate or clinically manifest indicators of chronic conditions in men that might contribute to the phenomenon "frailty". The evidence that reductions in endogenous androgens play a role in age-related health problems is circumstantial. Therefore, large-scale randomized trials are needed to establish whether aging males with low serum androgen levels benefit from androgen supplementation. PMID- 12890596 TI - Parental monitoring, negotiated unsupervised time, and parental trust: the role of perceived parenting practices in adolescent health risk behaviors. AB - PURPOSE: To compare two different parenting practices (parental monitoring and negotiated unsupervised time) and perceived parental trust in the reporting of health risk behaviors among adolescents. METHODS: Data were derived from 692 adolescents in 9th and 10th grades (x = 15.7 years) enrolled in health education classes in six urban high schools. Students completed a self-administered paper based survey that assessed adolescents' perceptions of the degree to which their parents monitor their whereabouts, are permitted to negotiate unsupervised time with their friends and trust them to make decisions. Using gender-specific multivariate logistic regression analyses, we examined the relative importance of parental monitoring, negotiated unsupervised time with peers, and parental trust in predicting reported sexual activity, sex-related protective actions (e.g., condom use, carrying protection) and substance use (alcohol, tobacco, and marijuana). RESULTS: For males and females, increased negotiated unsupervised time was strongly associated with increased risk behavior (e.g., sexual activity, alcohol and marijuana use) but also sex-related protective actions. In males, high parental monitoring was associated with less alcohol use and consistent condom use. Parental monitoring had no affect on female behavior. Perceived parental trust served as a protective factor against sexual activity, tobacco, and marijuana use in females, and alcohol use in males. CONCLUSIONS: Although monitoring is an important practice for parents of older adolescents, managing their behavior through negotiation of unsupervised time may have mixed results leading to increased experimentation with sexuality and substances, but perhaps in a more responsible way. Trust established between an adolescent female and her parents continues to be a strong deterrent for risky behaviors but appears to have little effect on behaviors of adolescent males. PMID- 12890597 TI - Examining the relationship between adolescent sexual risk-taking and perceptions of monitoring, communication, and parenting styles. AB - PURPOSE: To examine the relationship between adolescent sexual risk-taking and perception of parental monitoring, frequency of parent-adolescent communication, and parenting style. The influences of gender, age, and ethnicity are also of interest. METHODS: Data were collected from 7th-12th grade students in six rural, ethnically diverse school located in adjacent counties in a Southeastern state. A 174-item instrument assessed adolescent perceptions, behaviors and attitudes. Youth who had engaged in sexual intercourse (n = 1160) were included in the analyses. Logistic regression analyses were conducted to identify parenting practices that predicted high versus low-risk sex (defined by number of partners and use of condoms). Variables included parental monitoring, parent-adolescent communication, parenting style, parenting process interaction effects and interaction effects among these three parenting processes and gender, age and ethnicity. Analyses included frequencies, cross-tabulations and logistic regression. RESULTS: Parental monitoring, parental monitoring by parent adolescent communication and parenting style by ethnicity were significant predictors of sexual risk-taking. No gender or age interactions were noted. CONCLUSION: Parental monitoring, parent-adolescent communication and parenting style are all important variables to consider when examining sexual risk-taking among adolescents. PMID- 12890598 TI - Do adolescents want to avoid pregnancy? Attitudes toward pregnancy as predictors of pregnancy. AB - PURPOSE: To document the extent to which adolescents feel ambivalent towards getting pregnant and to examine the relationship between pregnancy attitudes and the occurrence of a pregnancy one year later. Demographic correlates of pregnancy attitudes also were examined. METHODS: This was a prospective study using a subsample of the National Longitudinal Study of Adolescent Health (Add Health) data base. The sample was 4869 adolescent females in grades 9 through 11 who completed two interviews at a one year interval. Logistic regression analyses were used to predict occurrence of a pregnancy at wave 2 from attitudes toward pregnancy at wave 1. RESULTS: A substantial number of adolescent females (15% 30%) reported some degree of ambivalence toward becoming pregnant relative to their peers. Adolescent females' attitudes towards pregnancy were predictive of the occurrence of a pregnancy one year later. Additionally, demographic correlates of the pregnancy attitude were identified, including differences due to ethnicity, age, relationship status, mother's education level, and whether the adolescent came from a one or a two parent home. CONCLUSIONS: Results suggest that a significant minority of adolescents have some ambivalence toward pregnancy relative to their peers and that these attitudes are predictive of the occurrence of pregnancy. PMID- 12890599 TI - Maternal discussions about pregnancy and adolescents, attitudes toward pregnancy. AB - PURPOSE: To identify aspects of maternal-adolescent relationships that are associated with high ambivalence toward pregnancy. METHODS: This was a cross sectional study of 350 inner-city, African-American females between the ages of 14 and 17 years residing in the Philadelphia metropolitan area. Adolescent females and their resident female caretaker (usually the biological mother) were both interviewed. Multiple regression analyses were used to predict adolescents' pregnancy attitudes from maternal-adolescent discussions about pregnancy, perceived maternal disapproval of pregnancy, and relationship satisfaction. RESULTS: Greater levels of discussion about negative consequences of pregnancy, stronger levels of perceived maternal disapproval, and higher levels of satisfaction all were associated with more negative attitudes towards pregnancy on the part of the adolescent. CONCLUSIONS: Maternal communication about the negative consequences of experiencing a pregnancy and the quality of the mother daughter relationship more generally may impact on adolescent attitudes toward pregnancy. PMID- 12890600 TI - Ethnic variation in parenting characteristics and adolescent smoking. AB - PURPOSE: To examine ethnic variation in parenting characteristics and in associations among parenting characteristics and adolescent smoking, and to assess the strength of those associations separately by ethnic subgroup. METHODS: Data were collected from a diverse sample (48.4% Hispanic/Latino, 22.9% Asian, 12.1% Non-Hispanic white, and 16.6% Multiethnic) of 1846 Southern California 6th graders (mean age 11.3 years). Lifetime smoking and parenting characteristics (parental smoking status, adolescents' perceptions of parent-child communication, and parental monitoring) were assessed. Across ethnic groups, descriptive statistics for parenting and smoking variables were calculated and compared. Logistic regression analyses controlled for demographics (socioeconomic status, generation status, family structure, age, and gender) and interaction terms (parenting characteristics x ethnicity), and were also run separately by ethnicity. RESULTS: Asian adolescents reported less parental monitoring than Latino/Hispanics and less parental communication than other ethnic subgroups. Odds ratios indicated parental monitoring (OR =.63; CI =.53-.74; p <.001) and parental communication (OR =.73; CI =.62-.86; p <.001) were protective whereas parental smoking (OR = 1.48; CI = 1.18-1.87; p <.01) was a risk factor for smoking. Relative to Latinos, parental monitoring (OR =.50; CI =.26-.95; p <.05) was more protective, and parental communication (OR = 2.44; CI = 1.15-5.17; p <.05) less protective for white adolescents. All parental characteristics were significantly associated with Latino/Hispanics smoking. Parental monitoring was significantly associated with Multiethnic and white smoking. No parenting characteristics were significant for Asians. CONCLUSION: Ethnic differences in parenting characteristics and adolescent smoking should be considered when designing prevention curriculum. Future research should investigate the nature of parent-child communication among Asian immigrant populations. PMID- 12890601 TI - The role of mother-daughter sexual risk communication in reducing sexual risk behaviors among urban adolescent females: a prospective study. AB - PURPOSE: To prospectively examine the relationship between mother-daughter communication about sex and selected sexual risk behaviors among inner-city adolescent females. METHODS: Participants were 219 sexually experienced females, 12 to 19 years of age, recruited from an inner-city adolescent medicine clinic in Philadelphia, PA, and randomly assigned to the control group of an HIV-risk reduction intervention study. Analyses were limited to data from control group participants to avoid confounding intervention effects. Poisson regression was employed to model three self-reported sexual risk behaviors: number of male sexual partners, number of episodes of sexual intercourse, and number of episodes of unprotected intercourse. Mediation effects were evaluated using variables from the Theory of Planned Behavior. Data were analyzed using Poisson regression. RESULTS: Higher levels of mother-daughter sexual risk communication were associated with fewer episodes of sexual intercourse and unprotected intercourse at 3-month follow-up. There was evidence that the relationship of communication to unprotected intercourse was mediated by condom use self-efficacy. Mother daughter sexual risk communication was not significantly associated with adolescents' reports of numbers of male sexual partner. CONCLUSIONS: This prospective study supports the notion that mothers who communicate with their daughters about sex can affect their daughters' sexual behaviors in positive ways. These findings lend support for the design and implementation of family based approaches to improve parent-adolescent sexual risk communication as one means of reducing HIV-related sexual risk behaviors among inner-city adolescent females. PMID- 12890602 TI - Relative influences of perceived parental monitoring and perceived peer involvement on adolescent risk behaviors: an analysis of six cross-sectional data sets. AB - PURPOSE: To assess: (a) the relative impact of monitoring and peer involvement among six cohorts of African-American youth in their mid-adolescent years, over a decade of research in one urban area, and (b) the consistency of the impact of the two influences over time. METHODS: Baseline data were collected from six cohorts involving 1279 low income African-American youth aged 13 to 16 years involved in community based studies conducted over a decade in an urban area. Self-reported behaviors, and perceptions of parental monitoring and peer risk involvement were assessed through structured questions. Data were analyzed by frequency distribution, one-way ANOVA, and multiple logistic regression. RESULTS: There was a rapid increase in sexual activity and substance use behaviors during mid-adolescence. Monitoring had a protective influence on substance use behaviors and sexual activity, but had no impact on condom use or drug trafficking. Peer involvement influenced all evaluated risk behaviors. The influences overall did not statistically change over time. CONCLUSIONS: Despite the marked increase in risk behaviors during mid-adolescence, monitoring and peer involvement both influenced adolescent behaviors across each cohort. PMID- 12890603 TI - Acculturation and parental attachment in Asian-American adolescents' alcohol use. AB - PURPOSE: To test whether the degree of acculturation predicts subsequent alcohol use among Asian-American adolescents, and to test the moderating effect of parental attachment. METHODS: This was a prospective study using a subsample of the National Longitudinal Adolescent Health data set. A nationally representative sample of 714 Asian-American boys (n = 332) and girls (n = 382) in grades 7-12 was analyzed. In-home self-report data were collected on two types of acculturation status, alcohol use, demographics, and parental attachment. After controlling for acculturation status and background variables at Wave I, logistic regression analysis was used to estimate the odds ratios to assess the association between acculturation and alcohol use at Wave II for adolescents. RESULTS: Asian-American adolescents with the highest level of acculturation (English use at home, born in the United States) were identified as the highest risk group. For adolescents with low parental attachment, the odds of alcohol use were 11 times greater in the highly acculturated group than in the least acculturated group. However, the odds of alcohol use for adolescents with moderate or high levels of parental attachment did not vary across acculturation groups. CONCLUSIONS: Overall, a greater level of acculturation was associated with greater alcohol use. However, when parental attachment was taken into account, highly acculturated adolescents with moderate or high parental attachment had no greater risk than adolescents with same levels of parental attachment who were less acculturated. Thus, it appears that acculturation per se was not a risk factor unless it was accompanied by a low level of parental attachment. PMID- 12890604 TI - Parental monitoring among adolescents in Beijing, China. PMID- 12890605 TI - Crossing the threshold. PMID- 12890606 TI - Comparison of hCMV immediate early and CaMV 35S promoters in both plant and human cells. AB - Cauliflower mosaic virus 35S promoter, widely used in transgenic crop plants, is known to be recognized in widely differing kinds of cells. Its activity in human cells may have impact on the risk assessment for the environmental release of genetically modified plants. In this study, transient expression of several constructs containing beta-glucuronidase (GUS) gene driven by cauliflower mosaic virus 35S promoter or by immediate early promoter of human cytomegalovirus (pCMV) was tested in both potato leaf protoplasts and cultured human cells. The results showed very low but measurable activity of 35S promoter in human 293T-cells (0.01% of that revealed when using pCMV) and in 293 cells that do not produce SV40 T antigen this activity was even lower. On the other hand, in potato protoplasts, pCMV displayed nearly 1% activity seen with p35S. PMID- 12890607 TI - Engineering cyclodextrin glycosyltransferase into a starch hydrolase with a high exo-specificity. AB - Cyclodextrin glycosyltransferase (CGTase) enzymes from various bacteria catalyze the formation of cyclodextrins from starch. The Bacillus stearothermophilus maltogenic alpha-amylase (G2-amylase is structurally very similar to CGTases, but converts starch into maltose. Comparison of the three-dimensional structures revealed two large differences in the substrate binding clefts. (i) The loop forming acceptor subsite +3 had a different conformation, providing the G2 amylase with more space at acceptor subsite +3, and (ii) the G2-amylase contained a five-residue amino acid insertion that hampers substrate binding at the donor subsites -3/-4 (Biochemistry, 38 (1999) 8385). In an attempt to change CGTase into an enzyme with the reaction and product specificity of the G2-amylase, which is used in the bakery industry, these differences were introduced into Thermoanerobacterium thermosulfurigenes CGTase. The loop forming acceptor subsite +3 was exchanged, which strongly reduced the cyclization activity, however, the product specificity was hardly altered. The five-residue insertion at the donor subsites drastically decreased the cyclization activity of CGTase to the extent that hydrolysis had become the main activity of enzyme. Moreover, this mutant produces linear products of variable sizes with a preference for maltose and had a strongly increased exo-specificity. Thus, CGTase can be changed into a starch hydrolase with a high exo-specificity by hampering substrate binding at the remote donor substrate binding subsites. PMID- 12890608 TI - Functional analysis of expressed peptides that bind yeast STE proteins. AB - Peptides are potentially useful for target validation and other reverse genetic applications. For instance, if a specific protein is susceptible to peptide inhibition, it may have a higher probability of being vulnerable to small molecules. We used the yeast two-hybrid technique to identify and study peptide binders for three yeast proteins involved in pheromone response: Ste11p, Ste18p, and Ste50p. A subset of peptide binders was shown to inhibit pheromone response in cells using two different functional assays. In addition, we utilized a variant of the yeast two-hybrid method to examine relative binding affinities based on competitive interactions in yeast. Our results suggest that binding affinity and inhibitory potency of peptides do not correlate perfectly and that peptide-protein interactions can be complex and unpredictable. Taken together these results suggest that while peptides are useful as in vivo inhibitors of protein function, caution must be exercised when choosing peptides for further studies and when inferring affinities from expression phenotypes. PMID- 12890609 TI - A new variant activator involved in the degradation of phenolic compounds from a strain of Pseudomonas putida. AB - A new variant type of regulatory activator and relevant promoters (designated capR, Pr and Po) involved in the metabolism of phenolic compounds were cloned from Pseudomonas putida KCTC1452 by using PCR. The deduced amino acid sequence of CapR revealed a difference in nine amino acids from the effector binding domain of DmpR. To measure effector specificity, plasmids were constructed in such a way that the expression of luc gene for firefly luciferase or lacZ for beta galactosidase as a reporter was under the control of capR. When Escherichia coli transformed with the plasmids was exposed to phenol, dramatic increases in the activity of luciferase or beta-galactosidase were observed in a range of 0.01-1 mM. Among various phenolic compounds tested, other effective compounds included catechol, 2-methylphenol, 3-methylphenol, 4-methylphenol, 2-chlorophenol, 4 chlorophenol, 2-nitrophenol, resorcinol, and 2, 5-dimethylphenol. The results indicate that CapR has effector specificity different from other related activators, CatR and DmpR. Waste water and soil potentially containing phenolic compounds were also tested by this system and the results were compared with chemical and GC data. The present results indicate that the biosensor consisting of capR and the promoters may be utilized for the development of a phenolic compounds-specific biosensor in monitoring the environmental pollutant. PMID- 12890610 TI - Integration of distributed multi-analyzer monitoring and control in bioprocessing based on a real-time expert system. AB - A computer system solution for integration of a distributed bioreactor monitoring and control instrumentation on the laboratory scale is described. Bioreactors equipped with on-line analyzers for mass spectrometry, near-infrared spectroscopy, electrochemical probes and multi-array gas sensors and their respective software were networked through a real-time expert systems platform. The system allowed data transmission of more than 1800 different signals from the instrumentation, including signals from gas sensors, electrodes, spectrometer detectors, balances, flowmeters, etc., and were used for processing and carrying out a number of computational tasks such as partial least-square regression, principal component analysis, artificial neural network modelling, heuristic decision-making and adaptive control. The system was demonstrated on different cultivations/fermentations which illustrated sensor fusion control, multivariate statistical process monitoring, adaptive glucose control and adaptive multivariate control. The performance of these examples showed high operational stability and reliable function and meet typical requirements for production safety and quality. PMID- 12890611 TI - Molecular engineering of biotin-glutamic acid decarboxylase 65 fusion protein (Biotin-GAD65) for non-radioactive GAD65 antibody assay. AB - We constructed biotinylated fusion proteins that linked to three detection tags to GAD65 at the N-terminus, and expressed them in an E. coli expression system. The Biotin14-GAD65 protein exhibited the strongest binding to both the GAD65 antibody and the streptavidin among the three constructs. We describe the optimal conditions using a Biotin14-GAD65-based immunoassay for the detection of GAD65 antibody. PMID- 12890612 TI - Identifying and modulating disulfide formation in the biopharmaceutical production of a recombinant protein vaccine candidate. AB - Structural conversion of the serotype A recombinant botulinum neurotoxin heavy chain fragment (rBoNTA(Hc)) produced intracellularly in Pichia pastoris yeast was observed and characterized during purification development efforts. A pH screening study captured the transformation stages of the original recovered species into its derived counterpart and a number of analytical tools such as peptide mapping by LC/MS confirmed the formation of a disulfide bond, especially in samples of neutral to basic pH. A cation exchange chromatographic method proved useful in following the incidence of the reaction in various rBoNTA(Hc) samples. The disulfide formation kinetics were characterized using a one-quarter quadratic factorial design, following the investigation and development of controlled oxidation conditions using cysteine and cystamine as the redox pair. Temperature, pH and concentration of the redox pair had a significant effect on the yield and rate of the disulfide formation. This controlled reaction was eventually introduced as a functional unit operation in the purification process. The summation of preliminary scale-up and potency data showed scalability and robustness in the production of an active disulfide-bonded form of a recombinant botulism vaccine candidate. The presence of the disulfide bond did not effect the vaccine potency and it enhanced the molecule's thermal stability. PMID- 12890613 TI - Bioorganosolve pretreatments for simultaneous saccharification and fermentation of beech wood by ethanolysis and white rot fungi. AB - Ethanol was produced by simultaneous saccharification and fermentation (SSF) from beech wood chips after bioorganosolve pretreatments by ethanolysis and white rot fungi, Ceriporiopsis subvermispora, Dichomitus squalens, Pleurotus ostreatus, and Coriolus versicolor. Beech wood chips were pretreated with the white rot fungi for 2-8 weeks without addition of any nutrients. The wood chips were then subjected to ethanolysis to separate them into pulp and soluble fractions (SFs). From the pulp fraction (PF), ethanol was produced by SSF using Saccharomyces cerevisiae AM12 and a commercial cellulase preparation, Meicelase, from Trichoderma viride. Among the four strains, C. subvermispora gave the highest yield on SSF. The yield of ethanol obtained after pretreatment with C. subvermispora for 8 weeks was 0.294 g g(-1) of ethanolysis pulp (74% of theoretical) and 0.176 g g(-1) of beech wood chips (62% of theoretical). The yield was 1.6 times higher than that obtained without the fungal treatments. The biological pretreatments saved 15% of the electricity needed for the ethanolysis. PMID- 12890614 TI - Enzymatic extraction of chondroitin sulfate from skate cartilage and concentration-desalting by ultrafiltration. AB - Skate cartilage is a fishery by-product, which contains chondroitin sulfate (CS), a glycosaminoglycan well known for its chondroprotective effect. Here described is a low-cost two-step process producing CS in non-denaturing conditions, consisting of an enzymatic extraction followed by tangential filtration to concentrate and purify CS. The performances of UF and MF membranes were compared in terms of flux and selectivity. The 0.1 microm-pore size membrane appeared to be the most efficient to separate CS from the other compounds. PMID- 12890615 TI - Human papillomavirus type 45 propagation, infection, and neutralization. AB - The organotypic (raft) culture system has allowed the study of the entire differentiation-dependent life cycle of human papillomaviruses (HPVs), including virion morphogenesis. We introduced linearized HPV45 genomic DNA into primary keratinocytes, where it recircularized and maintained episomally at a range of 10 50 copies of HPV genomic DNA. Following epithelial stratification and differentiation in organotypic culture, virion morphogenesis occurred. HPV45 virions were purified from raft cultures and were able to infect keratinocytes in vitro. By testing a panel of HPV VLP antisera, we were able to demonstrate that the infection was neutralized not only with human HPV45 VLP-specific antiserum, but also with human HPV18 VLP-specific antiserum, demonstrating serological cross reactivity between HPV18 and HPV45. PMID- 12890616 TI - Enhanced viral pathogenesis associated with a virulent mutant virus or a virulent satellite RNA correlates with reduced virion accumulation and abundance of free coat protein. AB - SatC, a chimeric satellite RNA associated with Turnip crinkle virus (TCV), intensifies the symptoms of TCV on all symptomatic hosts yet attenuates the symptoms of a TCV mutant that expresses low levels of a defective coat protein (CP). We now report that TCV virion levels were substantially reduced by the presence of satC or when two amino acids were inserted at the N-terminus of the CP, resulting in similarly enhanced symptoms. Since the TCV CP is a suppressor of RNA silencing, increased levels of resultant free CP could augment silencing suppression, resulting in enhanced colonization of the plant. PMID- 12890617 TI - Cellular expression of gH confers resistance to herpes simplex virus type-1 entry. AB - Entry of herpes simplex virus-1 (HSV-1) into cells requires a concerted action of four viral glycoproteins gB, gD, and gH-gL. Previously, cell surface expression of gD had been shown to confer resistance to HSV-1 entry. To investigate any similar effects caused by other entry glycoproteins, gB and gH-gL were coexpressed with Nectin-1 in Chinese hamster ovary (CHO) cells. Interestingly, cellular expression of gB had no effect on HSV-1(KOS) entry. In contrast, entry was significantly reduced in cells expressing gH-gL. This effect was further analyzed by expressing gH and gL separately. Cells expressing gL were normally susceptible, whereas gH-expressing cells were significantly resistant. Further experiments suggested that the gH-mediated interference phenomenon was not specific to any particular gD receptor and was also observed in gH-expressing HeLa cells. Moreover, contrary to a previous report, gL-independent cell surface expression of gH was detected in stably transfected CHO cells, possibly implicating cell surface gH in the interference phenomenon. Thus, taken together these findings indicate that cellular expression of gH interferes with HSV-1 entry. PMID- 12890618 TI - The cytoplasmic tails of infectious bronchitis virus E and M proteins mediate their interaction. AB - Virus-like particle (VLP) formation by the coronavirus E and M proteins suggests that interactions between these proteins play a critical role in coronavirus assembly. We studied interactions between the infectious bronchitis virus (IBV) E and M proteins using in vivo crosslinking and VLP assembly assays. We show that IBV E and M can be crosslinked to each other in IBV-infected and transfected cells, indicating that they interact. The cytoplasmic tails of both proteins are important for this interaction. We also examined the ability of the mutant and chimeric E and M proteins to form VLPs. IBV M proteins that are missing portions of their cytoplasmic tails or transmembrane regions were not able to support VLP formation, regardless of their ability to be crosslinked to IBV E. Interactions between the E and M proteins and the membrane bilayer are likely to play an important role in VLP formation and virus budding. PMID- 12890619 TI - The potato virus X TGBp2 protein association with the endoplasmic reticulum plays a role in but is not sufficient for viral cell-to-cell movement. AB - Potato virus X (PVX) TGBp1, TGBp2, TGBp3, and coat protein are required for virus cell-to-cell movement. Plasmids expressing GFP fused to TGBp2 were bombarded to leaf epidermal cells and GFP:TGBp2 moved cell to cell in Nicotiana benthamiana leaves but not in Nicotiana tabacum leaves. GFP:TGBp2 movement was observed in TGBp1-transgenic N. tabacum, indicating that TGBp2 requires TGBp1 to promote its movement in N. tabacum. In this study, GFP:TGBp2 was detected in a polygonal pattern that resembles the endoplasmic reticulum (ER) network. Amino acid sequence analysis revealed TGBp2 has two putative transmembrane domains. Two mutations separately introduced into the coding sequences encompassing the putative transmembrane domains within the GFP:TGBp2 plasmids and PVX genome, disrupted membrane binding of GFP:TGBp2, inhibited GFP:TGBp2 movement in N. benthamiana and TGBp1-expressing N. tabacum, and inhibited PVX movement. A third mutation, lying outside the transmembrane domains, had no effect on GFP:TGBp2 ER association or movement in N. benthamiana but inhibited GFP:TGBp2 movement in TGBp1-expressing N. tabacum and PVX movement in either Nicotiana species. Thus, ER association of TGBp2 may be required but not be sufficient for virus movement. TGBp2 likely provides an activity for PVX movement beyond ER association. PMID- 12890620 TI - The genome of bacteriophage phiKMV, a T7-like virus infecting Pseudomonas aeruginosa. AB - The complete DNA sequence of a new lytic T7-like bacteriophage phiKMV is presented. It is the first genome sequence of a member of the Podoviridae that infects Pseudomonas aeruginosa. The linear G + C-rich (62.3%) double-stranded DNA genome of 42,519 bp has direct terminal repeats of 414 bp and contains 48 open reading frames that are all transcribed from the same strand. Despite absence of homology at the DNA level, 11 of the 48 phiKMV-encoded putative proteins show sequence similarity to known T7-type phage proteins. Eighteen open reading frame products have been assigned, including an RNA polymerase, proteins involved in DNA replication, as well as structural, phage maturation, and lysis proteins. Surprisingly, the major capsid protein completely lacks sequence homology to any known protein. Also, the strong virulence toward many clinical P. aeruginosa isolates and a short replication time make phiKMV attractive for phage therapy or a potential source for antimicrobial proteins. PMID- 12890621 TI - Reduced expression of glutamate transporter EAAT2 and impaired glutamate transport in human primary astrocytes exposed to HIV-1 or gp120. AB - L-Glutamate is the major excitatory neurotransmitter in the brain. Astrocytes maintain low levels of synaptic glutamate by high-affinity uptake and defects in this function may lead to neuronal cell death by excitotoxicity. We tested the effects of HIV-1 and its envelope glycoprotein gp120 upon glutamate uptake and expression of glutamate transporters EAAT1 and EAAT2 in fetal human astrocytes in vitro. Astrocytes isolated from fetal tissues between 16 and 19 weeks of gestation expressed EAAT1 and EAAT2 RNA and proteins as detected by Northern blot analysis and immunoblotting, respectively, and the cells were capable of specific glutamate uptake. Exposure of astrocytes to HIV-1 or gp120 significantly impaired glutamate uptake by the cells, with maximum inhibition within 6 h, followed by gradual decline during 3 days of observation. HIV-1-infected cells showed a 59% reduction in V(max) for glutamate transport, indicating a reduction in the number of active transporter sites on the cell surface. Impaired glutamate transport after HIV-1 infection or gp120 exposure correlated with a 40-70% decline in steady-state levels of EAAT2 RNA and protein. EAAT1 RNA and protein levels were less affected. Treatment of astrocytes with tumor necrosis factor-alpha (TNF alpha) decreased the expression of both EAAT1 and EAAT2, but neither HIV-1 nor gp120 were found to induce TNF-alpha production by astrocytes. These findings demonstrate that HIV-1 and gp120 induce transcriptional downmodulation of the EAAT2 transporter gene in human astrocytes and coordinately attenuate glutamate transport by the cells. Reduction of the ability of HIV-1-infected astrocytes to take up glutamate may contribute to the development of neurological disease. PMID- 12890622 TI - Antigenic sites of coxsackie A9 virus inducing neutralizing monoclonal antibodies protective in mice. AB - A panel of murine IgG monoclonal antibodies (MAbs) was produced against coxsackievirus A9 (CAV9). Fifty-nine MAbs reactive in ELISA with purified CAV9 were identified. Eighteen of them could efficiently inhibit infection by CAV9 but not coxsackieviruses B. Neutralization-resistant CAV9 variants to four different MAbs were isolated and tested for resistance to neutralization by other MAbs of the panel. Three groups of reactivity including 10, 7, and 1 MAbs were thus identified. Sequencing of neutralization-escape virus mutants showed that neutralization by one MAb group was affected by change of VP3 amino acids 62 or 69. For the second group of reactivity, mutations included amino acids 154 or 165 of VP2. The only MAb of the third group selected for a change at residue 70 of VP2. Protection studies in a newborn mouse model of myositis suggested that either epitopes in VP2 or in VP3 mediate protection from CAV9 infection in vivo. PMID- 12890623 TI - High-throughput gene expression profiling indicates dysregulation of intestinal cell cycle mediators and growth factors during primary simian immunodeficiency virus infection. AB - During primary simian immunodeficiency virus (SIV) infection, CD4+ T cells are severely depleted in gut-associated lymphoid tissue (GALT), while CD8+ T-cell numbers dramatically increase. To gain an understanding of the molecular basis of this disruption in T-cell homeostasis, host gene expression was monitored in longitudinal jejunum tissue biopsies from SIV-infected rhesus macaques by DNA microarray analysis. Transcription of cyclin E1, CDC2, retinoblastoma, transforming growth factor (TGF), fibroblast growth factor (FGF), and interleukin 2 was repressed while cyclins B1 and D2 and transcription factor E2F were upregulated, indicating a complex dysregulation of growth and proliferation within the intestinal mucosa. Innate, cell-mediated, and humoral immune responses were markedly upregulated in animals that significantly reduced their viral loads and retained more intestinal CD4+ T cells. We conclude that the alterations in intestinal gene expression during primary SIV infection were characteristic of a broad-range immune response, and reflective of the efficacy of viral suppression. PMID- 12890624 TI - Relevance of Akt phosphorylation in cell transformation induced by Jaagsiekte sheep retrovirus. AB - Expression of the JSRV envelope (Env) is sufficient to transform immortalized rodent fibroblasts. A putative docking site for the PI3-K kinase (Y(590)-X-X M(593)) in the cytoplasmic tail of the transmembrane domain of the JSRV Env is a major determinant of viral-induced cell transformation. Akt is constitutively phosphorylated in rodent fibroblasts transformed by the JSRV Env. However, recent data suggest that Y590 and M593 are not necessary for JSRV Env-induced transformation of the immortalized chicken fibroblasts cell line DF-1. In this study we found that JSRV-induced transformation of DF-1 cells is Akt-independent. In addition, a replication-competent avian vector expressing the JSRV Env (RCASBP(A)+JE) was also able to induce transformation of primary chicken embryo fibroblasts (CEF). Vectors expressing JSRV Env Y590 mutants were still able to induce CEF cells transformation but not as efficiently as the vectors expressing the wild-type Env. In CEF cells, as in DF-1 cells, only the expression of the wild-type Env induced constitutive phosphorylation of Akt. Thus, in chicken cells, the degree of transformation induced by the JSRV Env is maximum in the presence of Y590 and Akt phosphorylation. We addressed the significance of Akt phosphorylation in rat 208F cells transformed by the JSRV Env and showed that Akt is indeed activated and shows kinase activity. Inhibitors of the PI-3K/Akt pathway reproducibly decreased the transformation efficiency of the JSRV Env. In vivo, we found phosphorylated Akt only in nasal tumors induced by the enzootic nasal tumor virus (ENTV), a JSRV-related beta-retrovirus. No evidence of Akt phosphorylation was found in lung tumor sections of sheep affected by pulmonary adenocarcinoma. As a whole, these results suggest that the activation of the PI 3K/Akt pathway contributes to the process of JSRV-induced cell transformation but most likely is not the primary determinant both in vitro and in vivo. PMID- 12890625 TI - Diversity of DNA beta, a satellite molecule associated with some monopartite begomoviruses. AB - DNA beta molecules are symptom-modulating, single-stranded DNA satellites associated with monopartite begomoviruses (family Geminiviridae). Such molecules have thus far been shown to be associated with Ageratum yellow vein virus from Singapore and Cotton leaf curl Multan virus from Pakistan. Here, 26 additional DNA beta molecules, associated with diverse plant species obtained from different geographical locations, were cloned and sequenced. These molecules were shown to be widespread in the Old World, where monopartite begomoviruses are known to occur. Analysis of the sequences revealed a highly conserved organization for DNA beta molecules consisting of a single conserved open reading frame, an adenine rich region, and a region of high sequence conservation [the satellite conserved region (SCR)]. The SCR contains a potential hairpin structure with the loop sequence TAA/GTATTAC; similar to the origins of replication of geminiviruses and nanoviruses. Two major groups of DNA beta satellites were resolved by phylogenetic analyses. One group originated from hosts within the Malvaceae and the second from a more diverse group of plants within the Solanaceae and Compositae. Within the two clusters, DNA beta molecules showed relatedness based both on host and geographic origin. These findings strongly support coadaptation of DNA beta molecules with their respective helper begomoviruses. PMID- 12890626 TI - Rhesus monkey rhadinovirus (RRV): construction of a RRV-GFP recombinant virus and development of assays to assess viral replication. AB - Rhesus monkey rhadinovirus (RRV) is a gamma-2-herpesvirus that is closely related to Kaposi's sarcoma-associated herpesvirus (KSHV/HHV-8). Lack of an efficient culture system to grow high titers of virus, and the lack of an in vivo animal model system, has hampered the study of KSHV replication and pathogenesis. RRV is capable of replicating to high titers on fibroblasts, thus facilitating the construction of recombinant rhadinoviruses. In addition, the ability to experimentally infect naive rhesus macaques with RRV makes it an excellent model system to study gamma-herpesvirus replication. Our study describes, for the first time, the construction of a GFP-expressing RRV recombinant virus using a traditional homologous recombination strategy. We have also developed two new methods for determining viral titers of RRV including a traditional viral plaque assay and a quantitative real-time PCR assay. We have compared the replication of wild-type RRV with that of the RRV-GFP recombinant virus in one-step growth curves. We have also measured the sensitivity of RRV to a small panel of antiviral drugs. The development of both the recombination strategy and the viral quantitation assays for RRV will lay the foundation for future studies to evaluate the contribution of individual genes to viral replication both in vitro and in vivo. PMID- 12890627 TI - First demonstration of a lack of viral sequence evolution in a nonprogressor, defining replication-incompetent HIV-1 infection. AB - It is universally acknowledged that genetic diversity is a hallmark of HIV-1 infection, and it is one of the traits that has considerably hampered the development of an effective vaccine. In a study of full-length HIV-1 genomic sequences (>9 kb), we show unique evidence for complete absence of viral evolution in an individual with truly nonprogressive infection. Gross gene defects were not detected, but the state of replication incompetence was attributed to the presence of stop codons in the structural genes gag p17 and p24 and in pol RT, which emerged as a consequence of G-A hypermutation. These inactivating mutations may have occurred early, soon after infection, during the clonal stage of primary viral replication, since these are the sole archival strains present today. This genetic homogeneity, with <1% variation between strains over an 8-year period, suggests that only limited proviral integration events occurred in this patient. Further study on the antigenic properties of this strain may assist in the development of HIV vaccines and therapeutics. PMID- 12890628 TI - Overlapping subdeletions within a 348-bp in the 5' exon of the LAT region that facilitates epinephrine-induced reactivation of HSV-1 in the rabbit ocular model do not further define a functional element. AB - A previous study identified a 348-bp region at the 5' end of the 8.5-kb latency associated transcript (LAT) of HSV-1 strain 17Syn+ that is necessary for maximum adrenergically induced reactivation following transcorneal iontophoresis of epinephrine (D.C. Bloom et al., 1996, J. Virol. 70, 2449-2459). In that study, the construct with complete deletion of the 348-bp region, 17delta348, failed to achieve the high reactivation frequency demonstrated by the parent (17Syn+) and rescued (17delta348R) viruses. To further characterize the function of the 348-bp region, we analyzed two genetic constructs with partial deletions in the same 348 bp region, 17delta201 and 17delta207, in the rabbit model. Both constructs exhibited the same high reactivation frequencies demonstrated by the parent 17Syn+ and the rescued 17delta348R viruses. These results suggest that the control of reactivation is distributed over a large portion of the 348-bp region, rather than being confined within a smaller, more discrete region. To assess whether the low reactivation phenotype of the 17delta348 construct was caused by a requirement for proper spacing of elements outside the 348-bp region, we constructed a virus (17delta348St) that contained a 360-bp stuffer fragment of heterologous DNA (lacZ) to maintain the proper spacing. The 17delta348St construct also displayed a low reactivation phenotype, similar to that of 17delta348, suggesting that the effect of deleting this segment of the 5' exon of LAT is obtained through a mechanism other than the disruption of spacing. PMID- 12890629 TI - Characterization of the low-copy HERV-Fc family: evidence for recent integrations in primates of elements with coding envelope genes. AB - In a previous search based on the envelope gene, we had identified two related proviral elements that could not be included in identified ERV families. An in silico database screening associated with an in vivo polymerase chain reaction search using primers in the reverse transcriptase domain, now allowed identification of a series of related elements, found at a limited number in simians. A phylogenetic analysis led to their inclusion in a new family of endogenous retroviruses with limited expansion, which we named ERV-Fc, and which is part of the enlarged ERV-F/H family. The human genome comprises only six HERV Fc, among which two possess full-length coding envelope genes. A complete provirus was identified in the baboon, also disclosing a fully open envelope gene. Cloning of the sites orthologous to the envelope-coding human proviruses demonstrated presence of the integrated proviruses in chimpanzee and gorilla, but not in orangutan. For the baboon element, the orthologous locus was found empty even in the phylogenetically most closely related macaque, again suggesting, together with the complete identity of its LTRs, "recent" integration. The data presented are compatible with an evolutionary scheme in which the ERV-Fc proviruses would be the endogenous traces of an active retroviral element, possibly acting as an infectious retrovirus with low endogeneization potency, with evidence for integrations at two distinct periods of primate evolution. PMID- 12890630 TI - Influence of CCR7 ligand DNA preexposure on the magnitude and duration of immunity. AB - The CC chemokine receptor (CCR) 7 ligands CCL21 and CCL19 were recently described as essential elements for establishing the microenvironment needed to initiate optimal immune responses in secondary lymphoid tissues. In the present study we have kinetically investigated the primary responses of naive DO11.10 TCR transgenic CD4+ T cells (OVA323-339 peptide specific) adoptively transferred into normal BALB/c mice given plasmid DNA encoding CCR7 ligands. The primary responses of CD4+ Tg-T cells in CCR7 ligand DNA recipients occurred more promptly, reaching levels higher than those observed in vector controls. In line with enhanced specific immunity, the T-cell population in CCR7 ligand recipients underwent more in vivo cell division following Ag stimulation, and a higher percentage of Ag specific T cells expressed an activation phenotype. Moreover, the enhanced primary responses of naive CD4+ T cells appeared to act via affects on migration and maturation of CD11c+ dendritic cells in the draining lymph nodes. In addition following mucosal challenge of herpes simplex virus-immune mice with virus, those that had received CCL21 or CCL19 during priming contained a higher frequency of responding CD4 T cells in lymph nodes and the site of infection. Moreover, CCL21- and CCL19-treated mice showed less severe disease and better survival following challenge. Our results are discussed in terms of the relevance of CCR7 ligand preimmunization to improve vaccine. PMID- 12890631 TI - Prior DNA immunization enhances immune response to dominant and subdominant viral epitopes induced by a fowlpox-based SIVmac vaccine in long-term slow-progressor macaques infected with SIVmac251. AB - A therapeutic vaccine for individuals infected with HIV-1 and treated with antiretroviral therapy (ART) should be able to replenish virus-specific CD4+ T cells and broaden the virus-specific CD8+ T-cell response in order to maintain CD8+ T-cell function and minimize viral immune escape after ART cessation. Because a combination of DNA and recombinant poxvirus vaccine modalities induces high levels of virus-specific CD4+ T-cell response and broadens the cytolytic activity in naive macaques, we investigated whether the same results could be obtained in SIVmac251-infected macaques. The macaques studied here were long-term nonprogressors that naturally contained viremia but were nevertheless treated with a combination of antiviral drugs to assess more carefully the effect of vaccination in the context of ART. The combination of a DNA expressing the gag and pol genes (DNA-SIV-gp) of SIVmac239 followed by a recombinant fowlpox expressing the same SIVmac genes (FP-SIV-gp) was significantly more immunogenic than two immunizations of FP-SIV-gp in SIVmac251-infected macaques treated with ART. The DNA/FP combination significantly expanded and broadened Gag-specific T cell responses measured by tetramer staining, ELISPOT, and intracellular cytokine staining and measurement of ex vivo cytolytic function. Importantly, the combination of these vaccine modalities also induced a sizeable expansion in most macaques of Gag-specific CD8-(CD4+) T-cells able to produce TNF-alpha. Hopefully, this modality of vaccine combination may be useful in the clinical management of HIV-1-infected individuals. PMID- 12890632 TI - Binding of ALX40-4C to APJ, a CNS-based receptor, inhibits its utilization as a co-receptor by HIV-1. AB - APJ, a G protein-coupled seven-transmembrane receptor, has been shown to serve as a co-receptor for the entry of human immunodeficiency virus type 1 (HIV-1), and it is dramatically expressed in central nervous system (CNS)-based cells. ALX40 4C was identified as a small-molecule antagonist of the chemokine receptor CXCR4, which can specifically inhibit HIV-1 entry via this co-receptor. In this study, we demonstrated that ALX40-4C inhibited both APJ- and CXCR4/APJ-mediated cell membrane fusion in a dose-dependent manner. In competitive binding assays, (125)I Apelin13 was replaced by ALX40-4C with an IC(50) of 2.9 microM, as compared with an IC(50) of 0.2 nM for Apelin13. Furthermore, ALX40-4C could block ligand induced APJ internalization and signaling. ALX40-4C, as an antagonist to APJ, directly binds to and prevents use of APJ as a HIV-1 co-receptor. Thus, ALX-4C has potential utility for further elucidation of HIV-1 neuropathogenesis and therapy of HIV-1-induced encephalopathy. PMID- 12890633 TI - Adenovirus type 5 DNA-protein complexes from formaldehyde cross-linked cells early after infection. AB - We report here the properties of viral DNA-protein complexes that purify with cellular chromatin following formaldehyde cross-linking of intact cells early after infection. The cross-linked viral DNA fractionated into shear-sensitive (S) and shear- resistant (R) components that were separable by sedimentation, which allowed independent characterization. The R component had the density and sedimentation properties expected for DNA-protein complexes and contained intact viral DNA. It accounted for about 50% of the viral DNA recovered at 1.5 h after infection but less than 20% by 4.5 h. The proportion of R component was independent of multiplicity of infection, even at less than one particle per cell. Viral hexon and protein VII, but not protein VI, were detected in the fractions containing the R component. These properties are consistent with those of partially uncoated virions associated with the nuclear envelope. A substantial proportion of the S component viral DNA had the same density as cellular chromatin. Protein VII was the most abundant viral protein present in gradient fractions that contained the S component. Complexes containing USF transcription factor cross-linked to the adenovirus major late promoter were detected by viral chromatin immunoprecipitation of the fractions containing S component. The S component probably contained uncoated nuclear viral DNA that assembles into early viral transcription complexes. PMID- 12890634 TI - Characterization of IgA response among women with incident HPV 16 infection. AB - Previous studies have characterized the prevalence and duration of serum IgG antibodies to human papillomavirus type 16 (HPV 16) in a well-studied cohort of college women, using viruslike particle- (VLP) based ELISAs. In this study IgA antibodies in cervical secretions and sera were examined using a newly developed capsomer-based ELISA and the patterns observed for serum IgG, serum IgA, and cervical IgA antibodies were compared. The median time to antibody detection from the first detection of HPV 16 DNA was 10.5 months for IgA in cervical secretions and 19.1 months for serum IgA. Serum IgA antibody conversion was observed less frequently and occurred later than IgA conversion in cervical secretions (P = 0.011) or serum IgG conversion (P = 0.051). The median time to antibody reversion, following seroconversion, was 12.0 months for IgA in cervical secretions and 13.6 months for serum IgA, whereas approximately 20% of women with serum IgG antibodies reverted within 36 months. Thus, the duration of IgA in cervical secretions and sera was shorter than the duration of serum IgG (P = 0.007 and 0.001). PMID- 12890635 TI - A trade-off in replication in mosquito versus mammalian systems conferred by a point mutation in the NS4B protein of dengue virus type 4. AB - An acceptable live-attenuated dengue virus vaccine candidate should have low potential for transmission by mosquitoes. We have identified and characterized a mutation in dengue virus type 4 (DEN4) that decreases the ability of the virus to infect mosquitoes. A panel of 1248 mutagenized virus clones generated previously by chemical mutagenesis was screened for decreased replication in mosquito C6/36 cells but efficient replication in simian Vero cells. One virus met these criteria and contained a single coding mutation: a C-to-U mutation at nucleotide 7129 resulting in a Pro-to-Leu change in amino acid 101 of the nonstructural 4B gene (NS4B P101L). This mutation results in decreased replication in C6/36 cells relative to wild-type DEN4, decreased infectivity for mosquitoes, enhanced replication in Vero and human HuH-7 cells, and enhanced replication in SCID mice implanted with HuH-7 cells (SCID-HuH-7 mice). A recombinant DEN4 virus (rDEN4) bearing this mutation exhibited the same set of phenotypes. Addition of the NS4B P101L mutation to rDEN4 bearing a 30 nucleotide deletion (Delta30) decreased the ability of the double-mutant virus to infect mosquitoes but increased its ability to replicate in SCID-HuH-7 mice. Although the NS4B P101L mutation decreases infectivity of DEN4 for mosquitoes, its ability to enhance replication in SCID HuH-7 mice suggests that it might not be advantageous to include this specific mutation in an rDEN4 vaccine. The opposing effects of the NS4B P101L mutation in mosquito and vertebrate systems suggest that the NS4B protein is involved in maintaining the balance between efficient replication in the mosquito vector and the human host. PMID- 12890636 TI - Establishment and maintenance of HSV latent infection is mediated through correct splicing of the LAT primary transcript. AB - To study the effect of the splicing of HSV-1 latency-associated transcript (LAT) on viral latency, we constructed two mutant viruses (FHlambda+ and FHlambda-) in which the 168-bp HpaI-HpaI fragment within the 2-kb LAT intron was replaced by a 447-bp bacteriophage lambda sequence. The lambda DNA was inserted in opposite orientations in FHlambda+ and FHlambda-. The mutation in FHlambda+ disrupted the splicing of LAT primary transcript and altered both LAT exon and intron, whereas the mutation in FHlambda- virus preserved the wild-type splice sites and the wild type exon. Quantitative PCR analysis revealed that during latency there was a reduction in the number of viral genomes in mouse trigeminal ganglia infected with FHlambda+ but not in those infected with FHlambda-. The decrease in the latent genome numbers was not due to a defect in viral replication during the acute stage of infection. Furthermore, trigeminal ganglia from mice latently infected with FHlambda+ displayed a slower reactivation kinetics compared to those infected with the parental strain. To elucidate the mechanism, we examined the antiapoptotic properties of these LAT constructs. A plasmid containing the pHlambda+ construct was found to be less protective for cells against apoptosis than plasmid containing the wild-type or pHlambda- constuct. These results suggest that the splicing of LAT primary transcript, and thus the correctly spliced exon product, play an important role in promoting the establishment and/or maintenance of viral latency. PMID- 12890637 TI - Hepatitis C virus core protein expression leads to biphasic regulation of the p21 cdk inhibitor and modulation of hepatocyte cell cycle. AB - Hepatitis C virus (HCV) Core protein is implicated in viral pathogenesis by the modulation of hepatocyte gene expression and function. To determine the effect of Core protein on the cell-cycle control of hepatocytes, a HepG2 cell line containing a Flag-tagged Core under the control of an inducible promoter was generated. Initial Core protein expression included the presence of unprocessed (191 aa) and processed (173 aa) forms of the Core proteins with the processed form becoming dominant later. Expression of the 191 aa form of Core protein corresponded to an increase in the expression of the p21, a decrease in cdk2 dependent kinase activity, and a decrease in the percentage of cells in S-phase along with an accumulation of cells in the G(0)/G(1) phase of the cell cycle. As the processed form accumulated, the p21 levels started to decline, suggesting that Core protein regulates p21 expression in a biphasic manner. These findings implicate Core protein in potentially modulating hepatocyte cell cycle differentially in the early stages of infection through biphasic regulation of p21 cdk kinase inhibitor. PMID- 12890638 TI - Vascular dementia: a diagnostic conundrum for the clinical neuropsychologist. AB - The contribution of cerebrovascular disease (CVD) to cognitive impairment in older persons is an important determination rendered by clinical neuropsychologists. However, it is often difficult to determine the relationship between CVD and severe cognitive dysfunction in this population. As a result, whether a patient meets diagnostic criteria for vascular dementia (VaD) or not is difficult to establish with reasonable clinical certainty. Our position is that five clinical myths regarding VaD complicate the diagnostic process for clinical neuropsychologists. Specifically, beliefs regarding a stepwise course of decline, patchy neuropsychological profile, prominent memory deficit, specificity of neuroimaging findings, and distinctions between VaD and Alzheimer's disease all represent factors that potentially confuse rather than clarify the diagnostic process. In this article we review these factors and offer suggestions for future research. PMID- 12890639 TI - Examination of "postconcussion-like" symptoms in a healthy sample. AB - The post-concussion syndrome (PCS) is relatively common following mild traumatic brain injury (MTBI). However, the factors that cause and maintain this syndrome continue to be debated. The purpose of this investigation was to examine the prevalence of postconcussion-like symptoms in a sample of healthy individuals. Participants (N = 104) completed the British Columbia Postconcussion Symptom Inventory-Short Form (BC-PSI-Sf), a test designed to measure both the frequency and intensity of ICD-10 criteria for PCS, and the Beck Depression Inventory (2nd ed.). Specific endorsement rates of postconcussion-like symptoms ranged from 35.9% to 75.7% for any experience of the symptoms in the past 2 weeks, and from 2.9% to 15.5% for the experience of more severe symptoms. Symptoms reported on the BC-PSI-Sf also showed a moderately high correlation with self-reported depressive symptoms [r (102) =.76, p <.01]. This study illustrates that the presence of postconcussion-like symptoms: (a). are not unique to mild head injury and are commonly found in healthy individuals, and (b). are highly correlated with depressive symptoms. PMID- 12890640 TI - Traumatic brain injury brief outcome interview. AB - There is much evidence that deficits in physical and psychological functioning persist long after traumatic brain injury occurs. This paper presents a brief outcome interview (BOI) that can be administered in person or over the telephone, with evaluation of change in functioning in three areas: (a). occupational status, (b). mobility/activities of daily living (ADL), and (c). social relationships. Forty-four traumatic brain injury participants were evaluated at an average of 6.2 years postinjury with the present BOI as well as with the Glasgow Outcome Scale and Karnofsky Performance Scale (KPS). The BOI demonstrated strong concurrent validity with both scales, as well as strong test-retest reliability. IQ and memory scores obtained at an average of 4.1 months postinjury suggested that the injury was moderately severe. The average score on the GPS suggested "good recovery" and the average score on the KPS suggested "normal activity with effort, some signs or symptoms." These descriptions matched the BOI for the mobility/ADL dimension, for which all respondents reported some form of independent mobility, and 88.6% of the respondents reported no need for any kind of assistance in daily life functioning. However, significant long-term issues were seen for social and occupational functioning. Fifty-four percent said that they did not socialize as much as before their injury, and half of the participants reported not being involved in a romantic relationship in spite of an average age of 32 years. In terms of occupational status, 40.9% reported not working at all at any kind of job. Compared to before their injury, 47.7% said this was less time, 40.9% said that it was for a lower salary, and 54.5% said that their responsibilities were less. The stability of these social and occupational changes was indicated by high test-test reliabilities for the overall BOI score and the three subscale scores (r's ranged from.97 to 1.0). These stable long-term changes are consistent with some other studies and suggest that the social and occupational needs of this population warrant more attention. PMID- 12890641 TI - Premorbid intellectual functioning, education, and brain size in traumatic brain injury: an investigation of the cognitive reserve hypothesis. AB - Cognitive reserve theories have been postulated in an attempt to explain individual differences in functional outcome following cerebral insult or disease. These theories suggest that higher education and psychometric intelligence may preserve functional capacity regardless of injury or disease severity. This study investigated cognitive reserve in 25 participants with traumatic brain injury (TBI) using high-resolution magnetic resonance imaging (MRI) analyses. We examined the relationships between total intracranial volume (TICV), ventricle-tobrain ratio (VBR), education level, and standardized testing obtained prior to injury with post-injury cognitive outcome. Participants with lower post-injury IQ scores had significantly lower TICV values, irrespective of injury severity, and experienced significantly greater change in IQ from pre- to post-injury. TICV and education correctly predicted participants' post-injury IQ category ( Y 90 or < 90). However, premorbid standardized testing (PST) scores did not predict cognitive outcome. The results of this study suggest that larger premorbid brain volume and higher education level may decrease vulnerability to cognitive deficits following TBI, consistent with the notion of a cognitive reserve. PMID- 12890642 TI - Clinical utility of the trail making test ratio score. AB - It has been suggested that a ratio score of part B to part A of the Trail Making Test (TMT) may have clinical utility as a measure of cognitive efficiency and/or dissimulation. We investigated the clinical utility of the TMT B/A ratio score in the evaluation of patients with traumatic brain injury. Part B of the TMT demonstrated a statistically significant difference between groups with mild (n = 59) versus moderate-severe (n = 41) injuries. However, the B/A ratio score did not demonstrate sensitivity to injury severity and also failed to discriminate reliably individuals who had independent psychometric indicators of possible dissimulation from those who did not. We conclude that in individuals with traumatic brain injury, the B/A ratio score does not appear to enhance the clinical utility of the TMT and that interpretation of performance on this test should instead involve comparing subtest completion times to established normative data. PMID- 12890643 TI - Quantitative EEG diagnostic confirmation of expressive aprosodia. AB - Quantitative electroencephalography (QEEG) was used to confirm predictions emanating from the neuropsychological examination of an adolescent with expressive aprosodia. Previous research has indicated that expressive deficits in prosody can be caused by cerebral damage to the right hemisphere region homologous to Broca's area. The neuropsychological evaluation of this individual indicates probable right anterior cerebral dysfunction. Taken together, a priori hypotheses included a relative deficit in cerebral arousal of right-relative to left-frontotemporal regions. EEG results confirm the hypothesis and provide case study evidence for both: (a). right anterior mediation of prosodic expression as well as (b). support for the inclusion of QEEG as part of a clinical neuropsychological evaluation. PMID- 12890644 TI - Symptoms and cognitive dysfunction in patients with the late whiplash syndrome. AB - The aim of the study was to record the severity of symptoms in patients with the late whiplash syndrome and to assess how the symptom load affected the performance on a simple psychometric test. The Rivermead Post-Concussion Symptoms Questionnaire (RPQ) with an additional question on neck pain, modified RPQ (mRPQ), and the Short Test of Mental Status (STMS) were applied on 40 chronic whiplash patients. Symptoms reported by the whiplash group were compared to those of 50 normal controls. The patients scored significantly higher than the controls on all symptoms listed in the mRPQ. The most frequent symptoms reported. PMID- 12890645 TI - A role for PYK2 in ANG II-dependent regulation of the PHAS-1-eIF4E complex by multiple signaling cascades in vascular smooth muscle. AB - Regulation of the PHAS-1-eukaryotic initiation factor-4E (eIF4E) complex is the rate-limiting step in the initiation of protein synthesis. This study characterized the upstream signaling pathways that mediate ANG II-dependent phosphorylation of PHAS-1 and eIF4E in vascular smooth muscle. ANG II-dependent PHAS-1 phosphorylation was maximal at 10 min (2.47 +/- 0.3 fold vs. control). This effect was completely blocked by the specific inhibitors of phosphatidylinositol 3-kinase (PI3-kinase, LY-294002), mammalian target of rapamycin, and extracellular signal-regulated kinase 1/2 (ERK1/2, U-0126) or by a recombinant adenovirus encoding dominant-negative Akt. PHAS-1 phosphorylation was followed by dissociation of eIF4E. Increased ANG II-induced eIF4E phosphorylation was observed at 45 min (2.63 +/- 0.5 fold vs. control), was maximal at 90 min (3.38 +/- 0.3 fold vs. control), and was sustained at 2 h. This effect was blocked by inhibitors of the ERK1/2 and p38 mitogen-activated protein (MAP) kinase pathways, but not by PI3-kinase inhibition, and was dependent on PKC, intracellular Ca2+, and tyrosine kinases. Downregulation of proline-rich tyrosine kinase 2 (PYK2) by antisense oligonucleotides led to a near-complete inhibition of PHAS-1 and eIF4E phosphorylation in response to ANG II. Therefore, PYK2 represents a proximal signaling intermediate that regulates ANG II-induced vascular smooth muscle cell protein synthesis via regulation of the PHAS-1-eIF4E complex. PMID- 12890646 TI - Regulation of elastin gene transcription by interleukin-1 beta-induced C/EBP beta isoforms. AB - We previously showed that interleukin (IL)-1beta decreases elastin gene transcription through activation of the NF-kappaB subunit p65 in neonatal rat lung fibroblasts. The present study was undertaken to further explore the molecular mechanisms responsible for the inhibitory effect of IL-1beta on elastin gene transcription. We found that cycloheximide blocked IL-1beta-induced downregulation of elastin mRNA but did not inhibit IL-1beta-induced translocation of p65 into the nucleus. IL-1beta treatment increased CCAAT/enhancer-binding protein (C/EBP)beta mRNA and protein levels including liver-enriched activating protein (LAP) and liver-enriched inhibitory protein (LIP), which was cycloheximide sensitive. C/EBPbeta isoforms bound a GCAAT-containing sequence in the proximal elastin promoter as determined by electrophoretic gel shift studies and confirmed by using specific anti-C/EBPbeta antibodies and by competition studies with oligonucleotides. Transient transfection of LIP expression vectors strongly decreased the transcriptional activity of the cotransfected elastin promoter and decreased levels of endogenous elastin mRNA. We demonstrated that IL 1beta-induced downregulation of elastin mRNA is dependent on NF-kappaB activation and C/EBPbeta expression. These results indicate that IL-1beta treatment activates NF-kappaB, which subsequently induces LIP expression and inhibition of elastin gene transcription. PMID- 12890647 TI - Distribution and functional properties of human KCNH8 (Elk1) potassium channels. AB - The Elk subfamily of the Eag K+ channel gene family is represented in mammals by three genes that are highly conserved between humans and rodents. Here we report the distribution and functional properties of a member of the human Elk K+ channel gene family, KCNH8. Quantitative RT-PCR analysis of mRNA expression patterns showed that KCNH8, along with the other Elk family genes, KCNH3 and KCNH4, are primarily expressed in the human nervous system. KCNH8 was expressed at high levels, and the distribution showed substantial overlap with KCNH3. In Xenopus oocytes, KCNH8 gives rise to slowly activating, voltage-dependent K+ currents that open at hyperpolarized potentials (half-maximal activation at -62 mV). Coexpression of KCNH8 with dominant-negative KCNH8, KCNH3, and KCNH4 subunits led to suppression of the KCNH8 currents, suggesting that Elk channels can form heteromultimers. Similar experiments imply that KCNH8 subunits are not able to form heteromultimers with Eag, Erg, or Kv family K+ channels. PMID- 12890648 TI - Apoptosis by Cd2+ or CdMT in proximal tubule cells: different uptake routes and permissive role of endo/lysosomal CdMT uptake. AB - The mechanisms of cadmium-metallothionein (CdMT) uptake and toxicity in proximal tubule (PT) cells are not well understood. The effects of 10 microM CdCl2 or Cd7MT-1 (MT-1 saturated with 10 microM CdCl2) on 109Cd2+ uptake, viability, and MT levels of cultured rat PT cells were investigated. Apical 109Cd2+ uptake was measured in confluent monolayers, apoptosis was assessed with Hoechst 33342, and intracellular MT levels were monitored by immunofluorescence and quantitative morphometry. 109Cd2+ uptake into PTC increased over time and plateaued at 24 h. 109Cd7MT-1 uptake was delayed but reached a similar magnitude after 40 h. With Cd2+, apoptosis occurred within 4 h, peaked at 24 h, and declined at 48-72 h. Cd7MT-1 induced apoptosis after 24-36 h, reaching similar levels as with Cd2+ after 48 h. Cd2+ and Cd7MT-1 significantly increased intracellular MT immunoreactivity after 20 and 4 h, respectively. The weak base chloroquine and the inhibitor of phosphatidylinositol 3-kinases, LY-294002, selectively inhibited the effects of Cd7MT-1 on MT immunoreactivity and apoptosis. PT cells accumulated 109Cd7MT-1 in membrane vesicles associated with the late endo/lysosomal marker LAMP1 but less with the early endosomal marker Rab5a, which was abolished by chloroquine or LY-294002. Thus development of apoptosis followed the uptake kinetics of Cd2+ and Cd7MT-1. Endo/lysosomal inhibitors prevented uptake of Cd7MT 1 into endo/lysosomes and apoptosis but had no effect on these parameters with Cd2+, suggesting that apoptosis of PT cells is triggered by free cytosolic Cd2+, either by direct apical transport or by translocation of free Cd2+ from endo/lysosomes after endocytosis of Cd7MT-1. PMID- 12890649 TI - K+ depolarization induces RhoA kinase translocation to caveolae and Ca2+ sensitization of arterial muscle. AB - KCl causes smooth muscle contraction by elevating intracellular free Ca2+, whereas receptor stimulation activates an additional mechanism, termed Ca2+ sensitization, that can involve activation of RhoA-associated kinase (ROK) and PKC. However, recent studies support the hypothesis that KCl may also increase Ca2+ sensitivity. Our data showed that the PKC inhibitor GF-109203X did not, whereas the ROK inhibitor Y-27632 did, inhibit KCl-induced tonic (5 min) force and myosin light chain (MLC) phosphorylation in rabbit artery. Y-27632 also inhibited BAY K 8644- and ionomycin-induced MLC phosphorylation and force but did not inhibit KCl-induced Ca2+ entry or peak ( approximately 15 s) force. Moreover, KCl and BAY K 8644 nearly doubled the amount of ROK colocalized to caveolae at 30 s, a time that preceded inhibition of force by Y-27632. Colocalization was not inhibited by Y-27632 but was abolished by nifedipine and the calmodulin blocker trifluoperazine. These data support the hypothesis that KCl caused Ca2+ sensitization via ROK activation. We discuss a novel model for ROK activation involving translocation to caveolae that is dependent on Ca2+ entry and involves Ca2+-calmodulin activation. PMID- 12890650 TI - Alteration in expression of myosin isoforms in detrusor smooth muscle following bladder outlet obstruction. AB - Partial urinary bladder outlet obstruction (PBOO) in men, secondary to benign prostatic hyperplasia, induces detrusor smooth muscle (DSM) hypertrophy. However, despite DSM hypertrophy, some bladders become severely dysfunctional (decompensated). Using a rabbit model of PBOO, we found that although DSM from sham-operated bladders expressed nearly 100% of both the smooth muscle myosin heavy chain isoform SM-B and essential light chain isoform LC17a, DSM from severely dysfunctional bladders expressed as much as 75% SM-A and 40% LC17b (both associated with decreased maximum velocity of shortening). DSM from dysfunctional bladder also exhibited tonic-type contractions, characterized by slow force generation and high force maintenance. Immunofluorescence microscopy showed that decreased SM-B expression in dysfunctional bladders was not due to generation of a new cell population lacking SM-B. Metabolic cage monitoring revealed decreased void volume and increased voiding frequency correlated with overexpression of SM A and LC17b. Myosin isoform expression and bladder function returned toward normal upon removal of the obstruction, indicating that the levels of expression of these isoforms are markers of the PBOO-induced dysfunctional bladders. PMID- 12890651 TI - Galpha12 regulates epithelial cell junctions through Src tyrosine kinases. AB - Regulation and assembly of the epithelial cell junctional complex involve multiple signaling mechanisms, including heterotrimeric G proteins. Recently, we demonstrated that Galpha12 binds to the tight junction scaffolding protein ZO-1 through the SH3 domain and that activated Galpha12 increases paracellular permeability in Madin-Darby canine kidney (MDCK) cells (Meyer et al. J Biol Chem 277: 24855-24858, 2002). In the present studies, we explore the effects of Galpha12 expression on tight and adherens junction proteins and examine downstream signaling pathways. By confocal microscopy, we detect disrupted tight and adherens junction proteins with increased actin stress fibers in constitutively active Galpha12 (QLalpha12)-expressing MDCK cells. The normal distribution of ZO-1 and Na-K-ATPase was altered in QLalpha12-expressing MDCK cells, consistent with loss of polarity. We found that the tyrosine kinase inhibitor genistein and the Src-specific inhibitor PP-2 reversibly abrogated the QLalpha12 phenotype on the junctional complex. Junctional protein localization was preserved in PP-2- or genistein-treated QLalpha12-expressing cells, and the increase in paracellular permeability as measured by transepithelial resistance and [3H]mannitol flux was prevented by the inhibitors. Src activity was increased in QLalpha12-expressing MDCK cells as assessed by Src autophosphorylation, and beta-catenin tyrosine phosphorylation was also increased, although there was no detectable increase in Rho activity. Taken together, these results indicate that Galpha12 regulates MDCK cell junctions, in part through Src tyrosine kinase pathways. PMID- 12890652 TI - Occupational asthma: research, change and the 30% target. PMID- 12890653 TI - Scientific principles and pragmatic solutions for the measurement of exposure to inhalable dust. AB - In order to understand and control the risks presented by inhalation of dust at work, research over many years has been focused on understanding how dust present in the air enters the human nose and mouth during the act of breathing. For health-related dust exposure monitoring, sampling devices are needed that collect the same 'inhalable fraction' of dust as the human head. Mark and Vincent's 1986 paper presented a study that has contributed more than any other to the practical realization of this inhalable dust concept. The authors developed a simple solution--the IOM personal inhalable dust sampler--to what we now know is an extremely complex problem. Although scientific understanding has grown in the years since this paper was published, very few other dust sampling instruments have emerged as being able to meet both the scientific criteria and the practical need for inhalable dust measurement. Both authors have continued to build on this work and have made further contributions to our theoretical and practical understanding in this field. PMID- 12890654 TI - Technical, occupational health and environmental aspects of metal degreasing with aqueous cleaners. AB - Aqueous cleaners used for metal degreasing are detergent formulations containing surfactants (such as linear alkylbenzene sulphonates, alkylphenol ethoxylates or alcohol ethoxylates), builders (such as hydroxides, phosphates or silicates), sequestrants (such as EDTA or NTA), anti-corrosive agents (such as ethanolamines), solvents (such as glycol ethers or d-limonene) and other specialty additives. Generally sold as concentrates, they are typically diluted between 3 and 20 times in water, leading to solutions containing only a few per cent active products. The cleaning efficiency depends on physicochemical phenomena such as wetting, solubilization, emulsification, dispersion, sequestration and saponification, and is enhanced by thermal and mechanical energy. Cleaning equipment is based on spraying or immersion of the parts and may include drying and rinsing steps. Because of the complexity and variability of the mixtures, the occupational health and environmental evaluation of aqueous cleaners is based on the study of their components. Aqueous cleaners are generally believed to present a low risk to workers' health and to the environment. However, some anionic surfactants and strong alkalis are skin and eye irritants, ethanolamines are allergenic and several glycol ethers of the ethylene glycol family are proven systemic toxicants that are easily absorbed through the skin. Although most components of aqueous cleaners are biodegradable and of low ecotoxicity, alkylphenol ethoxylates degrade into persistent and toxic compounds. Phosphates, if released directly into the environment, may cause eutrophication of rivers and lakes. Waste recycling or treatment by specialized facilities is usually required for spent solutions containing contaminants such as oils and heavy metals. From a technical, toxicological and environmental standpoint, aqueous cleaners can be used successfully to replace traditional organic solvents used in metal degreasing. PMID- 12890655 TI - Expert judgment and occupational hygiene: application to aerosol speciation in the nickel primary production industry. AB - In many situations characterized by sparse data, occupational hygienists have used subjective judgments that are claimed to be derived from their experience and knowledge. While this practice is widespread, there has been no systematic study of 'expert judgment' or the 'art' of occupational hygiene. Indeed, there is a need to address the question of whether there is such a thing as 'expert opinion' in occupational hygiene that is broadly shared by practicing professionals. This research, employing 11 experts who estimate an exposure parameter (the percentages of four nickel species) in 12 workplaces in a nickel primary production industry, provides a large dataset from which useful inferences can be drawn about the quality of expert judgments and the variability among the experts. A well-designed questionnaire that provided succinct information about the processes and baseline data served to calibrate the experts. The Bayesian framework has been used in this work to develop posterior means and standard deviations of the percentages of the four nickel species in the 12 workplaces of interest in the company. These estimates of the nickel speciation are at least as precise as--and most of the time more precise than- those provided by the sparse measurement data. There was a very high degree of agreement among the experts. A majority of the experts agreed among themselves 92% of the time, while almost two-thirds agreed 73% of the time. This, coupled with the fact that the experts came from varied backgrounds, seems to suggest that there is indeed some broad body of specialized knowledge that the experts are drawing on to reach similar judgments. It also seems that one type of expert is not necessarily any better than any other kind, and expertise does not necessarily require intimate familiarity with the workplace. In this example, the expert judgment exercise has indeed enhanced the quality of our knowledge of the exposure 'fingerprints' for the nickel industry workplaces studied and the combination of expert judgment and sparse data is better than the sparse data alone. For occupational hygiene exposure assessment, our experience suggests that such expert judgment methods can provide a cost-effective means to improve and refine information about workplace hazards. However, more study is warranted for situations where the domain of the quantity of interest has a much wider range of values, e.g. actual exposure values. PMID- 12890656 TI - Validation of a semi-quantitative job exposure matrix at a Soderberg aluminum smelter. AB - OBJECTIVES: We tested the validity of a job exposure matrix (JEM) for coal tar pitch volatiles (CTPV) at a Soderberg aluminum smelter. The JEM had been developed by a committee of company hygienists and union representatives for an earlier study of cancer incidence and mortality. Our aim was to test the validity and reliability of the expert-based assignments. METHODS: Personal CTPV exposure measurements (n = 1879) overlapped 11 yr of the JEM. The arithmetic mean was calculated for 35 job/time period combinations (35% of the exposed work history), categorized using the original exposure intervals, and compared with the expert based assignments. RESULTS: The expert-based and the measurement-based exposure assignments were only moderately correlated (Spearman's rho = 0.42; weighted kappa = 0.39, CI 0.10-0.69). Only 40% of the expert-based medium category assignments were correctly assigned, with better agreement in the low (84%) and high (100%) categories. Pot operation jobs exhibited better agreement (rho = 0.60) than the maintenance and pot shell repair jobs (rho = 0.25). The mid-point value of the medium category was overestimated by 0.3 mg/m(3). CONCLUSIONS: The expert-based exposure assignments may be improved by better characterizing the transitions between exposure categories, by accounting for exposure differences between pot lines and by re-examining the category mid-point values used in calculating the cumulative exposure. Lack of historical exposure measurements often requires reliance on expert knowledge to assess exposure levels. Validating the experts' estimates against available exposure measurements may help to identify weaknesses in the exposure assessment where improvements may be possible, as was shown here. PMID- 12890657 TI - Occupational characteristics of cases with asbestos-related diseases in The Netherlands. AB - OBJECTIVE: To describe the occupational background of cases with an asbestos related disease and to present overall mesothelioma risks across industries with historical exposure to asbestos. METHODS: For the period 1990-2000, cases were collected from records held by two law firms. Information on jobs held, previous employers, activities performed and specific products used were obtained from patients themselves or next of kin. Branches of industry and occupations were coded and the likelihood of asbestos exposure was assessed. For each branch of industry, the overall risk of mesothelioma was calculated from the ratio of the observed number of mesothelioma cases and the cumulative population-at-risk in the period 1947-1960. In order to compare mesothelioma risks across different industries, risk ratios were calculated for the primary asbestos industry and asbestos user industries relative to all other branches of industry. RESULTS: In total, 710 mesotheliomas and 86 asbestosis cases were available. The average latency period was approximately 40 yr and the average duration of exposure was 22 yr. Ship building and maintenance contributed the largest number of cases (27%), followed by the construction industry (14%), the insulation industry (12%), and the navy and army, primarily related to ship building and maintenance (5%). In the insulation industry, the overall risk of mesothelioma was 5 out of 100 workers, and in the ship building industry, 1 out of 100 workers. The construction industry had an overall risk comparable with many other asbestos using industries (7 per 10,000 workers), but due to its size claimed many mesothelioma cases. CONCLUSION: The majority of cases with asbestos-related diseases had experienced their first asbestos exposure prior to 1960. For cases with first asbestos exposure after 1960, a shift was observed from the primary asbestos industry towards asbestos-using industries, such as construction, petroleum refining, and train building and maintenance. Due to the long latency period, asbestos exposure from 1960 to 1980 will cause a considerable number of mesothelioma cases in the next two decades. PMID- 12890658 TI - Occupational toluene exposure and auditory function: results from a follow-up study. AB - The ototoxicity of occupational exposure to toluene below 50 p.p.m. was investigated in a longitudinal study over 5 yr with four repeated examinations starting with 333 male workers from rotogravure printing plants. Past lifetime weighted average exposures (LWAE) to toluene and noise were determined from individual work histories; recent individual exposures were measured 10 times during the study (toluene, active sampling; noise, stationary measurements). The auditory thresholds were measured with pure tone audiometry. The mean LWAE exposures to toluene and noise were 45 +/- 17 p.p.m. plus 82 +/- 7 dB(A) for printers (high toluene intensity) and 10 +/- 7 p.p.m. plus 82 +/- 4 dB(A) for end processors (low toluene intensity). The mean current exposures to toluene and noise during the study were 26 +/- 20 p.p.m. plus 81 +/- 4 dB(A) for printers and 3 +/- 3 p.p.m. plus 82 +/- 4 dB(A) for end-processors. Repeated measurement analyses (grouping factors: toluene intensity, exposure duration and noise intensity) and logistic regressions did not reveal significant effects of toluene intensity, of exposure duration and of interactions between toluene intensity and noise intensity. The stratification dependent on noise intensity itself [79 +/- 3 versus 84 +/- 1 dB(A)] was significantly associated with the auditory thresholds. Regarding the missing toluene effects, it was concluded that the threshold level for developing a hearing loss as a result of occupational toluene exposure might be above the actual limit of 50 p.p.m. PMID- 12890659 TI - Biological monitoring for selected herbicide biomarkers in the urine of exposed custom applicators: application of mixed-effect models. AB - Metabolites and/or parent compounds of the herbicides atrazine, alachlor, metolachlor, cyanazine and the 2-ethylhexyl ester of 2,4-dichlorophenoxyacetic acid (2,4-D) were measured in the urine of 15 custom applicators who each provided from five to seven 24 h urine samples during a 6 week period (n = 87). Each applicator provided a pre-season urine sample and a reference population (n = 46) provided first-morning urine samples. Urinary biomarkers were measured by either immunoassay or gas chromatography. During the spraying season, the geometric mean amount of alachlor mercapturate equivalents (eq.), atrazine eq., 2,4-D and metolachlor mercapturate eq. excreted in 24 h was 17, 19, 110 and 22 nmol, respectively. Mixed-effect models were used to determine predictors of the amount of atrazine eq. and 2,4-D excreted in 24 h. The specific days of herbicide spraying associated with increased biomarker excretion varied for the two analytes, and included one or more days prior to urine collection. This confirms the importance of collecting covariate information on day(s) most relevant to the biomarker of interest. The within-worker variance component, expressed as a geometric standard deviation ((W)GSD range: 2.5-2.9), was substantially larger than the between-worker component ((B)GSD range: 1.3-1.5) for the modeled biomarkers. Alachlor mercapturate eq. and metolachlor mercapturate eq. were detected in more than half of the applicator pre-season urine samples. All biomarkers were detected infrequently in the reference population. Evaluation of non-spray exposure determinants was limited by inclusion of prior day spraying, adjustment for time and the small sample size. PMID- 12890660 TI - IOHA/ICOH Declaration on Occupational Hygiene. PMID- 12890661 TI - Composition and function of PDZ protein complexes during cell polarization. AB - Complexes consisting of PDZ proteins have been implicated in a variety of cellular processes. In recent years, it has become increasingly clear that PDZ proteins play essential roles during the establishment of spatial asymmetry in various metazoan cell types such as epithelial cells. Epithelial cells possess asymmetry with respect to the apicobasal axis reflected by the differential distribution of proteins and lipids in the apical and basolateral surfaces. In Drosophila, three PDZ protein complexes have been shown to play crucial functions during the establishment of cell-cell adhesions and epithelial cell polarity: Bazooka/Dm-Par6/DaPKC, Crumbs/Stardust/Discs Lost, and Scribble/Discs Large/Lethal Giant Larvae. In this review, we focus primarily on our current knowledge of the localization and function of these complexes in Drosophila epithelia. We also discuss recent data that enhance our understanding of the homologous protein complexes and their roles during junctional assembly and polarization of mammalian epithelial cells. PMID- 12890662 TI - Role of Na-K-ATPase in the assembly of tight junctions. AB - Na-K-ATPase, also known as the sodium pump, is a crucial enzyme that regulates intracellular sodium homeostasis in mammalian cells. In epithelial cells Na-K ATPase function is also involved in the formation of tight junctions through RhoA GTPase and stress fibers. In this review, a new two-step model for the assembly of tight junctions is proposed: step 1, an E-cadherin-dependent formation of partial tight junction strands and of the circumferential actin ring; and step 2, active actin polymerization-dependent tethering of tight junction strands to form functional tight junctions, an event requiring normal function of Na-K-ATPase in epithelial cells. A new role for stress fibers in the assembly of tight junctions is proposed. Also, implications of Na-K-ATPase function on tight junction assembly in diseases such as cancer, ischemia, hypomagnesemia, and polycystic kidney disease are discussed. PMID- 12890663 TI - Inhibition of heme oxygenase decreases sodium and fluid absorption in the loop of Henle. AB - We previously demonstrated that carbon monoxide (CO) stimulates the apical 70-pS K+ channel in the thick ascending limb (TAL) of the rat kidney (Liu HJ, Mount DB, Nasjletti A, and Wang WH. J Clin Invest 103: 963-970, 1999). Because the apical K+ channel plays a key role in K+ recycling, we tested the hypothesis that heme oxygenase (HO)-dependent metabolites of heme may affect Na+ transport in the TAL. We used in vivo microperfusion to study the effect of chromium mesoporphyrin (CrMP), an inhibitor of HO, on fluid absorption (Jv) and Na+ absorption (JNa) in the loop of Henle and renal clearance methods to examine the effect of CrMP on renal sodium excretion. Microperfusion experiments demonstrated that addition of CrMP to the loop of Henle decreased Jv by 13% and JNa by 20% in animals on normal rat chow and caused a decrease in Jv (39%) and JNa (40%) in rats on a high-K+ (HK) diet. The effect of CrMP is the result of inhibition of HO because addition of MgPP, an analog of CrMP that does not inhibit HO, had no effect on Jv. Western blot analysis showed that HO-2 is expressed in the kidney and that the level of HO-2 was significantly elevated in animals on a HK diet. Renal clearance studies demonstrated that the infusion of CrMP increased the excretion of urinary Na+ (ENa) and volume (UV) without changes in glomerular filtration rate. The effect of CrMP on ENa and UV was larger in HK rats than those kept on normal chow. We conclude that HK intake increases HO-2 expression in the kidney and that HO dependent metabolites of heme, presumably CO, play a significant role in the regulation of Na+ transport in the loop of Henle. PMID- 12890664 TI - Acute application of TNF stimulates apical 70-pS K+ channels in the thick ascending limb of rat kidney. AB - TNF has been shown to be synthesized by the medullary thick ascending limb (mTAL) (21). In the present study, we used the patch-clamp technique to study the acute effect of TNF on the apical 70-pS K+ channel in the mTAL. Addition of TNF (10 nM) significantly stimulated activity of the 70-pS K+ channel and increased NPo [a product of channel open probability (Po) and channel number (N)] from 0.20 to 0.97. The stimulatory effect of TNF was observed only in cell-attached patches but not in excised patches. Moreover, addition of TNF had no effect on the ROMK like small-conductance K+ channels in the TAL. The dose-response curve of the TNF effect yielded a Km value of 1 nM, a concentration that increased channel activity to 50% maximal stimulatory effect of TNF. The concentrations required for reaching the plateau of the TNF effect were between 5 and 10 nM. The stimulatory effect of TNF on the 70-pS K+ channel was observed in the presence of N(omega)-nitro-L-arginine methyl ester. This indicated that the effect of TNF was not mediated by a nitric oxide-dependent pathway. Also, inhibition of PKA did not affect the stimulatory effect of TNF. In contrast, inhibition of protein tyrosine kinase not only increased activity of the 70-pS K+ channel but also abolished the effect of TNF. Moreover, inhibition of protein tyrosine phosphatase (PTP) blocked the stimulatory effect of TNF on the 70-pS K+ channel. The notion that the TNF effect results from stimulation of PTP activity is supported by PTP activity assay in which treatment of mTAL cells with TNF significantly increased the activity of PTP. We conclude that TNF stimulates the 70-pS K+ channel via stimulation of PTP in the mTAL. PMID- 12890665 TI - [3H]Inulin as a marker for glomerular filtration rate. PMID- 12890666 TI - Crystal structure of carbapenam synthetase (CarA). AB - Carbapenam synthetase (CarA) is an ATP/Mg2+-dependent enzyme that catalyzes formation of the beta-lactam ring in (5R)-carbapenem-3-carboxylic acid biosynthesis. CarA is homologous to beta-lactam synthetase (beta-LS), which is involved in clavulanic acid biosynthesis. The catalytic cycles of CarA and beta LS mediate substrate adenylation followed by beta-lactamization via a tetrahedral intermediate or transition state. Another member of this family of ATP/Mg2+ dependent enzymes, asparagine synthetase (AS-B), catalyzes intermolecular, rather than intramolecular, amide bond formation in asparagine biosynthesis. The crystal structures of apo-CarA and CarA complexed with the substrate (2S,5S)-5 carboxymethylproline (CMPr), ATP analog alpha,beta-methyleneadenosine 5' triphosphate (AMP-CPP), and a single Mg2+ ion have been determined. CarA forms a tetramer. Each monomer resembles beta-LS and AS-B in overall fold, but key differences are observed. The N-terminal domain lacks the glutaminase active site found in AS-B, and an extended loop region not observed in beta-LS or AS-B is present. Comparison of the C-terminal synthetase active site to that in beta-LS reveals that the ATP binding site is highly conserved. By contrast, variations in the substrate binding pocket reflect the different substrates of the two enzymes. The Mg2+ coordination is also different. Several key residues in the active site are conserved between CarA and beta-LS, supporting proposed roles in beta-lactam formation. These data provide further insight into the structures of this class of enzymes and suggest that CarA might be a versatile target for protein engineering experiments aimed at developing improved production methods and new carbapenem antibiotics. PMID- 12890667 TI - Candida tropicalis expresses two mitochondrial 2-enoyl thioester reductases that are able to form both homodimers and heterodimers. AB - Here we report on the cloning of a Candida tropicalis gene, ETR2, that is closely related to ETR1. Both genes encode enzymatically active 2-enoyl thioester reductases involved in mitochondrial synthesis of fatty acids (fatty acid synthesis type II) and respiratory competence. The 5'- and 3'-flanking (coding) regions of ETR2 and ETR1 are about 90% (97%) identical, indicating that the genes have evolved via gene duplication. The gene products differ in three amino acid residues: Ile67 (Val), Ala92 (Thr), and Lys251 (Arg) in Etr2p (Etr1p). Quantitative PCR analysis and reverse transcriptase-PCR indicated that both genes were expressed about equally in fermenting and ETR1 predominantly respiring yeast cells. Like the situation with ETR1, expression of ETR2 in respiration-deficient Saccharomyces cerevisiae mutant cells devoid of Ybr026p/Etr1p was able to restore growth on glycerol. Triclosan that is used as an antibacterial agent against fatty acid synthesis type II 2-enoyl thioester reductases inhibited growth of FabI overexpressing mutant yeast cells but was not able to inhibit respiratory growth of the ETR2- or ETR1-complemented mutant yeast cells. Resolving of crystal structures obtained via Etr2p and Etr1p co-crystallization indicated that all possible dimer variants occur in the same asymmetric unit, suggesting that similar dimer formation also takes place in vivo. PMID- 12890668 TI - JUMONJI, a critical factor for cardiac development, functions as a transcriptional repressor. AB - JUMONJI (JMJ) is a nuclear factor that is critical for normal cardiovascular development, evidenced by the analysis of jmj homozygous mutant mice. However, the molecular function of JMJ remains to be elucidated. In the present study, we investigated whether JMJ is a transcriptional modulator. Reporter gene assays using the GAL4-DNA binding domain fused to JMJ and a reporter gene consisting of the GAL4 binding sites upstream of a luciferase reporter gene indicated that JMJ functions as a powerful transcriptional repressor. The DNA binding motif of JMJ was determined using CASTing experiments by incubating a random oligonucleotide library with the GST-JMJ fusion protein coupled to agarose beads. Among the selected binding oligonucleotides, the high affinity DNA binding sequences were identified by gel retardation assays. JMJ repressed expression of the reporter genes containing the high affinity JMJ binding sequences, indicating that JMJ is a DNA-binding transcriptional repressor. The domains for transcriptional repression, DNA binding, and nuclear localization signal were mapped by mutational analyses using reporter gene assays, gel retardation assays, and immunostaining experiments, respectively. The present data demonstrate for the first time that JMJ functions as a DNA-binding transcriptional repressor. Therefore, JMJ may play a critical role in transcription factor cascade to regulate expression of heart-specific genes and normal cardiac development. PMID- 12890670 TI - Protein kinase C alpha phosphorylates and negatively regulates diacylglycerol kinase zeta. AB - Diacylglycerol kinase (DGK) terminates diacylglycerol (DAG) signaling by phosphorylating DAG to produce phosphatidic acid, which also has signaling properties. Thus, precise control of DGK activity is essential for proper signal transduction. We demonstrated previously that a peptide corresponding to the myristoylated alanine-rich C kinase substrate (MARCKS) phosphorylation site domain (PSD) in DGK zeta was phosphorylated in vitro by an active fragment of protein kinase C (PKC). In the present study, we tested full-length DGK zeta and found that PKC alpha phosphorylated DGK zeta on serines within the MARCKS PSD in vitro and in vivo. DGK zeta also coimmunoprecipitated with PKC alpha, suggesting that they reside in a regulated signaling complex. We then tested whether phosphorylation affected DAG kinase activity. We found that a mutant (DGK zeta S/D) in which serines within the MARCKS PSD were altered to aspartates (to mimic phosphorylation) had lower activity compared with wild-type DGK zeta or a control mutant (DGK zeta S/N) in which the same serines were changed to asparagines. Furthermore, activation of PKC alpha by phorbol 12-myristate 13-acetate inhibited the activity of wild-type DGK zeta, but not DGK zeta S/D, in human embryonic kidney 293 cells. These results suggest that by phosphorylating the MARCKS PSD, PKC alpha attenuates DGK zeta activity. Supporting this, we found that cells expressing DGK zeta S/D had higher DAG levels and grew more rapidly compared with cells expressing DGK zeta S/N that could not be phosphorylated. Taken together, these results indicate that PKC alpha phosphorylates DGK zeta in cells, and this phosphorylation inhibits its kinase activity to remove cellular DAG, thereby affecting cell growth. PMID- 12890669 TI - Early growth response gene 1 modulates androgen receptor signaling in prostate carcinoma cells. AB - The transcription factor early growth response gene 1 (EGR1) has been implicated in diverse roles in the regulation of cell growth, apoptosis, and differentiation. Previous studies suggest that the effects of EGR1 on tumorigenesis are critically dependent on the cellular context. In a majority of prostate cancers, EGR1 is overexpressed and promotes prostate tumor progression. In contrast, in other tumor types such as breast cancers and glioblastomas, EGR1 is expressed at low levels and when overexpressed can inhibit tumor growth. To explore the role of EGR1 in prostate tumorigenesis, we examined the impact of EGR1 expression on the androgen receptor (AR) signaling pathway. We show here that EGR1 binds to the AR in prostate carcinoma cells, and an EGR1-AR complex can be detected by chromatin immunoprecipitation at the enhancer of an endogenous AR target gene. Overexpression of EGR1 enhanced AR-mediated transactivation, whereas EGR1 knockdown by small interfering RNA inhibited AR signaling pathway activity. Furthermore, Western blot and immunocytochemical analyses showed that constitutive overexpression of EGR1 promotes the translocation of AR from the cytoplasm to the nucleus. These results indicate that EGR1 may promote prostate cancer development by modulating the androgen receptor signaling pathway. PMID- 12890671 TI - The phosphatase MKP1 is a transcriptional target of p53 involved in cell cycle regulation. AB - The tumor suppressor p53 protein suppresses cell growth by inducing cell cycle arrest or apoptosis. Despite the fact that p53-dependent p21-mediated G1 arrest induced by DNA damage is well defined, the role of p53 in the cell cycle in response to the MAKP signaling remains to be determined. Here we show that MKP1, a member of the dual specificity protein phosphatase family capable of inactivating MAPKs, is a transcriptional target of p53. MKP1 mRNA and protein levels were increased upon p53 activation in several well defined p53-regulated cell systems. p53 bound to a consensus p53 binding site located in the second intron of the MKP1 gene and transactivated MKP1 in reporter gene assays. Inhibition of phosphatase activity impaired p53-mediated G1 arrest in arrested human glioblastoma GM cells in response to growth factor stimuli. Importantly conditional expression of MKP1 prevented arrested human cancer cells from entering into the cell cycle. Thus, these results provide a novel mechanism by which p53 controls the cell cycle in response to the MAPK signaling in the absence of DNA damage and suggest that p53 may negatively control the MAKP pathway via MKP1. PMID- 12890672 TI - NeuroD1/E47 regulates the E-box element of a novel zinc finger transcription factor, IA-1, in developing nervous system. AB - IA-1 is a novel zinc finger transcription factor with a restricted tissue distribution in the embryonic nervous system and tumors of neuroendocrine origin. The 1.7-kilobase 5'-upstream DNA sequence of the human IA-1 gene directed transgene expression predominantly in the developing nervous system including forebrain, midbrain, hindbrain, spinal cord, retina, olfactory bulb, and cerebellum, which recapitulated the expression patterns of neuroendocrine tissues and childhood brain tumors. The IA-1 promoter deletion reporter gene constructs revealed that the sequence between -426 and -65 bp containing three putative E boxes (approximately 361 bp) upstream of the transcription start site was sufficient to confer tissue-specific transcriptional activity. Further mutation analysis revealed that the proximal E-box (E3) closest to the start site is critical to confer transcriptional activity. Electrophoretic mobility shift assay and transient transfection studies demonstrated that the NeuroD1 and E47 heterodimer are the key transcription factors that regulate the proximal E-box of the IA-1 promoter. Therefore, we concluded that the IA-1 gene is developmentally expressed in the nervous system and the NeuroD1/E47 transcription factors up regulate IA-1 gene expression through the proximal E-box element of the IA-1 promoter. PMID- 12890673 TI - UUUUUNU stimulation of vaccinia virus early gene transcription termination. Oligonucleotide sequence and structural requirements for stimulation of premature termination in vitro. AB - Vaccinia virus early genes are unique in that transcription terminates in a signal- and factor-dependent manner. Recent results from this laboratory demonstrated that a 22-mer RNA oligonucleotide containing a central U9 sequence exhibited sequence- and concentration-dependent stimulation of premature transcription termination and transcript release in trans. In an effort to better understand the different aspects of the U5NU stimulation of premature termination, we evaluated the activity of various oligonucleotides in vitro. Neither RNA containing a mutant U5NU signal nor single-stranded DNA containing T5NT was able to stimulate premature termination, demonstrating both sequence specificity and a requirement for ribose. Furthermore, neither oligonucleotide was able to compete with U5NU, demonstrating that each failed to bind to the U5NU recognition factor. Substitution of the U9 signal with either BrU9 or BrdU9 inhibited normal termination but did not stimulate premature termination. The addition of BrdU5NdU inhibited U5NU stimulation of premature termination, demonstrating that both oligonucleotides bind to the same site on the U5NU recognition factor. Finally, U5NU containing RNA as short as nine bases served as an effective stimulator of premature termination. These observations impact directly on the development of oligonucleotide based anti-poxvirus therapeutic agents. PMID- 12890674 TI - Dissection of the contribution of individual domains to the ATPase mechanism of Hsp90. AB - Hsp90 is a dimeric, ATP-regulated molecular chaperone. Its ATPase cycle involves the N-terminal ATP binding domain (amino acids (aa) 1-272) and, in addition, to some extent the middle domain (aa 273-528) and the C-terminal dimerization domain (aa 529-709). To analyze the contribution of the different domains and the oligomeric state on the progression of the ATPase cycle of yeast Hsp90, we created deletion constructs lacking either the C-terminal or both the C-terminal and the middle domain. To test the effect of dimerization on the ATPase activity of the different constructs, we introduced a Cys residue at the C-terminal ends of the constructs, which allowed covalent dimerization. We show that all monomeric constructs tested exhibit reduced ATPase activity and a decreased affinity for ATP in comparison with wild type Hsp90. The covalently linked dimers lacking only the C-terminal domain hydrolyze ATP as efficiently as the wild type protein. Furthermore, this construct is able to trap the ATP molecule similar to the full-length protein. This demonstrates that in the ATPase cycle, the C terminal domain can be replaced by a cystine bridge. In contrast, the ATPase activity of the artificially linked N-terminal domains remains very low and bound ATP is not trapped. Taken together, we show that both the dimerization of the N terminal domains and the association of the N-terminal with the middle domain are important for the efficiency of the ATPase cycle. These reactions are synergistic and require Hsp90 to be in the dimeric state. PMID- 12890675 TI - Akt activity negatively regulates phosphorylation of AMP-activated protein kinase in the heart. AB - In the heart, insulin stimulates a variety of kinase cascades and controls glucose utilization. Because insulin is able to activate Akt and inactivate AMP activated protein kinase (AMPK) in the heart, we hypothesized that Akt can regulate the activity of AMPK. To address the potential existence of this novel signaling pathway, we used a number of experimental protocols to activate Akt in cardiac myocytes and monitored the activation status of AMPK. Mouse hearts perfused in the presence of insulin demonstrated accelerated glycolysis and glucose oxidation rates as compared with non-insulin-perfused hearts. In addition, insulin caused an increase in Akt phosphorylation and a decrease in AMPK phosphorylation at its major regulatory site (threonine 172 of the alpha catalytic subunit). Transgenic mice overexpressing a constitutively active mutant form of Akt1 displayed decreased phosphorylation of cardiac alpha-AMPK. Isolated neonatal cardiac myocytes infected with an adenovirus expressing constitutively active mutant forms of either Akt1 or Akt2 also suppressed AMPK phosphorylation. However, Akt-dependent depression of alpha-AMPK phosphorylation could be overcome in the presence of the AMPK activator, metformin, suggesting that an override mechanism exists that can restore AMPK activity. Taken together, this study suggests that there is cross-talk between the AMPK and Akt pathways and that Akt activation can lead to decreased AMPK activity. In addition, our data suggest that the ability of insulin to inhibit AMPK may be controlled via an Akt-mediated mechanism. PMID- 12890676 TI - Expression of the yeast PIS1 gene requires multiple regulatory elements including a Rox1p binding site. AB - The PIS1 gene is required for de novo synthesis of phosphatidylinositol (PI), an essential phospholipid in Saccharomyces cerevisiae. PIS1 gene expression is unusual because it is uncoupled from the other phospholipid biosynthetic genes, which are regulated in response to inositol and choline. Relatively little is known about regulation of transcription of the PIS1 gene. We reported previously that PIS1 transcription is sensitive to carbon source. To further our understanding of the regulation of PIS1 transcription, we carried out a promoter deletion analysis that identified three regions required for PIS1 gene expression (upstream activating sequence (UAS) elements 1-3). Deletion of either UAS1 or UAS2 resulted in an approximately 45% reduction in expression, whereas removal of UAS3 yielded an 84% decrease in expression. A comparison of promoters among several Saccharomyces species shows that these sequences are highly conserved. Curiously, the UAS3 element region (-149 to -138) includes a Rox1p binding site. Rox1p is a repressor of hypoxic genes under aerobic growth conditions. Consistent with this, we have found that expression of a PIS1-cat reporter was repressed under aerobic conditions, and this repression was dependent on both Rox1p and its binding site. Furthermore, PI levels were elevated under anaerobic conditions. This is the first evidence that PI levels are affected by regulation of PIS1 transcription. PMID- 12890677 TI - Ceramide enhances cholesterol efflux to apolipoprotein A-I by increasing the cell surface presence of ATP-binding cassette transporter A1. AB - It is widely accepted that functional ATP-binding cassette transporter A1 (ABCA1) is critical for the formation of nascent high density lipoprotein particles. However, the cholesterol pool(s) and the cellular signaling processes utilized by the ABCA1-mediated pathway remain unclear. Sphingomyelin maintains a preferential interaction with cholesterol in membranes, and its catabolites, especially ceramide, are potent signaling molecules that could play a role in ABCA1 regulation or function. To study the potential role of ceramide in this process, we treated a variety of cell lines with 20 microM C2-ceramide and examined apolipoprotein-mediated cholesterol efflux to lipid-free apoA-I. We found that cell lines expressing ABCA1 displayed 2-3-fold increases in cholesterol efflux to apoA-I. Cell lines not expressing ABCA1 were unaffected by ceramide. We further characterized the cholesterol efflux effect in Chinese hamster ovary cells. Ceramide treatment did not cause significant cytotoxicity or apoptosis and did not affect cholesterol efflux to non-apolipoprotein acceptors. Raising endogenous ceramide levels increased cholesterol efflux to apoA-I. Using a cell surface biotinylation method, we found that the total cellular ABCA1 and that at the plasma membrane were increased with ceramide treatment. Also ceramide enhanced the binding of fluorescently labeled apoA-I to Chinese hamster ovary cells. These data suggest that ceramide may increase the plasma membrane content of ABCA1, leading to increased apoA-I binding and cholesterol efflux. PMID- 12890678 TI - Activation of p53 by oxidative stress involves platelet-derived growth factor beta receptor-mediated ataxia telangiectasia mutated (ATM) kinase activation. AB - Phosphorylation of the p53 tumor suppressor protein is a critical event in the up regulation and activation of p53 during cellular stress. In this study, we characterized the signaling pathway linking oxidative stress to p53 through the platelet-derived growth factor beta (PDGF beta) receptor and the ataxia telangiectasia mutated (ATM) kinase. In response to H2O2, we observed phosphorylation of p53 specifically at serine 15, but not serine 9, 20, or 392. Phosphorylation of Ser-15 was correlated with enhanced induction and functional activation of p53 manifest as transcription of the p53 target p21CIP/WAF. We found that H2O2 induced phosphorylation of the PDGF beta receptor and increased ATM kinase activity, two events integral to p53 activation as either AG1433 (a PDGF beta receptor inhibitor) or caffeine (an ATM kinase inhibitor) inhibited Ser 15 phosphorylation. Similarly, p53 activation by H2O2 was inhibited by kinase inactive forms of the PDGF beta receptor or ATM. Inhibition of ATM kinase had no effect on H2O2-induced PDGF beta receptor tyrosine phosphorylation, whereas PDGF beta receptor suppression with RNA interference impaired H2O2-induced ATM activation, indicating that ATM lies downstream to the PDGF beta receptor in this signaling cascade. Functionally, inhibition of the PDGF beta receptor abrogated the inhibition of cell proliferation, and promotion of apoptosis due to H2O2 treatment. Thus, these data link PDGF beta receptor transactivation to H2O2 induced p53 phosphorylation and suggest a functional role for growth factor receptors in modulation of p53 function. PMID- 12890679 TI - Roles for homotypic interactions and transautophosphorylation in IkappaB kinase beta IKKbeta) activation [corrected]. AB - The nuclear factor kappaB (NF-kappaB)/Rel family of transcription factors participates in a wide range of biological activities including inflammation, immunity, and apoptosis. NF-kappaB is kept inactive in the cytoplasm in unstimulated cells by virtue of the masking of its nuclear localization sequence by bound IkappaB protein. Cellular stimuli trigger the destruction of IkappaB proteins and the liberation of NF-kappaB to enter the nucleus and activate gene expression. A multisubunit IkappaB kinase complex (IKK) phosphorylates IkappaB proteins and mediates the activation of NF-kappaB by proinflammatory stimuli such as tumor necrosis factor alpha. Phosphorylation of IkappaB proteins triggers their polyubiquitination and their subsequent recognition and degradation by the proteasome. The IKK complex contains two catalytic subunits, IKKalpha and IKKbeta, and a noncatalytic subunit, NF-kappaB essential modifier/IKKgamma. IKK activation depends upon the phosphorylation of residues in the activation loop of IKKbeta and the subsequent activation of IKKbeta kinase activity. However, the events contributing to IKKbeta phosphorylation are not well understood. Here, we present evidence that the activation of IKKbeta depends on its ability to form homotypic interactions and to transautophosphorylate. We find that an intact leucine zipper in IKKbeta is necessary for homotypic interactions, kinase activation, and phosphorylation on its activation loop. Enforced oligomerization of an IKKbeta mutant defective in forming homotypic interactions restores kinase activation. Homotypic interactions allow IKKbeta molecules to transautophosphorylate one another on their activation loops. Finally, the oligomerization of IKKbeta is stimulated by tumor necrosis factor alpha in cultured cells. Our findings support a model whereby ligand-induced homotypic interactions between IKKbeta molecules result in IKKbeta phosphorylation and consequently IKK activation. PMID- 12890680 TI - An Sp1-NF-Y/progesterone receptor DNA binding-dependent mechanism regulates progesterone-induced transcriptional activation of the rabbit RUSH/SMARCA3 gene. AB - Steroids regulate alternative splicing of rabbit RUSH/SMARCA3, an SWI/SNF-related transcription factor. Transactivation was evaluated in 2057 bp of genomic sequence. Truncation analysis identified a minimal 252-bp region with strong basal promoter activity in transient transfection assays. The size of the 5' untranslated region (233 bp) and the transcription start site were determined by primer extension analysis. The transcription start site mapped to a consensus initiator (Inr) element in a TATA-less region with a downstream promoter element (+29). These elements were authenticated by mutation/deletion analysis. The Inr/downstream promoter element combination is conserved in the putative core promoter (-35/+35) of the human ortholog, suggesting that transcription initiation is similarly conserved. Two Sp1 sites that are also conserved in the putative promoter of human SMARCA3 and a RUSH binding site (-616/-611) that is unique to the rabbit promoter repress basal transcription. These sites were variously authenticated by gel shift and chromatin immunoprecipitation assays. Analysis of the proximal promoter showed the -162/+90 region was required for progesterone responsiveness in transient transfection assays. Subsequent mutation/deletion analysis revealed a progesterone receptor half-site mediated induction by progesterone. An overlapping Y-box (in the reverse ATTGG orientation) repressed basal transcription and progesterone-induced transcriptional activation in the presence of the Sp1 sites. The specificity of progesterone receptor and transcription factor NF-Y binding were authenticated by gel shift assays. Chromatin immunoprecipitation assays confirmed the Y-box effects were mediated in a DNA binding-dependent fashion. This represents a unique regulatory scenario in which ligand-dependent transactivation by the progesterone receptor is subject to Sp1/NF-Y repression. PMID- 12890681 TI - Aggrecan protects cartilage collagen from proteolytic cleavage. AB - The matrix components responsible for cartilage mechanical properties, type II collagen and aggrecan, are degraded in osteoarthritis through proteolytic cleavage by matrix metalloproteinases (MMPs) and aggrecanases, respectively. We now show that aggrecan may serve to protect cartilage collagen from degradation. Although collagen in freeze-thawed cartilage depleted of aggrecan was completely degraded following incubation with MMP-1, collagen in cartilage with intact aggrecan was not. Using interleukin-1-stimulated bovine nasal cartilage explants where aggrecan depletion occurs during the first week of culture, followed by collagen loss during the second week, we evaluated the effect of selective MMP and aggrecanase inhibitors on degradation. A selective MMP inhibitor did not block aggrecan degradation but caused complete inhibition of collagen breakdown. Similar inhibition was seen with inhibitor addition following aggrecan depletion on day 6-8, suggesting that MMPs are not causing significant collagen degradation prior to the second week of culture. Inclusion of a selective aggrecanase inhibitor blocked aggrecan degradation, and, in addition, inhibited collagen degradation. When the inhibitor was introduced following aggrecan depletion, it had no effect on collagen breakdown, ruling out a direct effect through inhibition of collagenase. These data suggest that aggrecan plays a protective role in preventing degradation of collagen fibrils, and that an aggrecanase inhibitor may impart overall cartilage protection. PMID- 12890682 TI - Involvement of phospholipase D2 in lysophosphatidate-induced transactivation of platelet-derived growth factor receptor-beta in human bronchial epithelial cells. AB - Lysophosphatidate (LPA) mediates multiple cellular responses via heterotrimeric G protein coupled LPA-1, LPA-2, and LPA-3 receptors. Many G protein-coupled receptors stimulate ERK following tyrosine phosphorylation of growth factor receptors; however, the mechanism(s) of transactivation of receptor tyrosine kinases are not well defined. Here, we provide evidence for the involvement of phospholipase D (PLD) in LPA-mediated transactivation of platelet-derived growth factor receptor-beta (PDGF-R beta). In primary cultures of human bronchial epithelial cells (HBEpCs), LPA stimulated tyrosine phosphorylation of PDGF-R beta and threonine/tyrosine phosphorylation of ERK1/2. The LPA-mediated activation of ERK and tyrosine phosphorylation of PDGF-R beta was attenuated by tyrphostin AG 1296, an inhibitor of PDGF-R kinase, suggesting transactivation of PDGF-R by LPA. Furthermore, LPA-, but not PDGF beta-chain homodimer-induced tyrosine phosphorylation of PDGF-R beta was partially blocked by pertussis toxin, indicating coupling of LPA-R(s) to Gi. Exposure of HBEpCs to LPA activated PLD. Butan-1-ol, which acts as an acceptor of phosphatidate generated by the PLD pathway, blocked LPA-mediated transactivation of PDGF-R beta. This effect was not seen with butan-3-ol, suggesting PLD involvement. The role of PLD1 and PLD2 in the PDGF-R beta transactivation by LPA was investigated by infection of cells with adenoviral constructs of wild type and catalytically inactive mutants of PLD. LPA activated both PLD1 and PLD2 in HBEpCs; however, infection of cells with cDNA for wild type PLD2, but not PLD1, increased the tyrosine phosphorylation of PDGF-R beta in response to LPA. Also, the LPA-mediated tyrosine phosphorylation of PDGF-R beta was attenuated by the catalytically inactive mutant mPLD2-K758R. Infection of HBEpCs with adenoviral constructs of wild type hPLD1, mPLD2, and the inactive mutants of hPLD1 and mPLD2 resulted in association of PLD2 wild type and inactive mutant proteins with the PDGF-R beta compared with PLD1. These results show for the first time that transactivation of PDGF-R beta by LPA in HBEpCs is regulated by PLD2. PMID- 12890683 TI - Transcriptional regulation of mouse delta-opioid receptor gene by CpG methylation: involvement of Sp3 and a methyl-CpG-binding protein, MBD2, in transcriptional repression of mouse delta-opioid receptor gene in Neuro2A cells. AB - Opioid receptors are expressed in a cell type-specific manner. Here we show that the mouse delta-opioid receptor (mDOR) gene is regulated by promoter region CpG methylation. The mDOR promoter containing a putative CpG island is highly methylated in Neuro2A cells, correlating with the repression of this gene in these cells. This is in contrast with the unmethylated state of the mDOR promoter in NS20Y cells, which express a high level of mDOR. Repression of mDOR transcription in Neuro2A cells could be partially relieved by chemically induced demethylation with 5-aza-2'-deoxycytidine. In addition, in vitro methylation of the luciferase reporter gene driven by the mDOR promoter resulted in an inhibition of transcription in NS20Y cells. Methyl-CpG-binding protein complex 1 (MeCP1) has been implicated in methylation-mediated transcriptional repression of several genes. Electrophoretic mobility shift assays showed that fully methylated, but not unmethylated, mDOR promoter fragment formed a MeCP1-like protein complex that contained methyl-CpG-binding domain protein 2 (MBD2) and Sp3. Furthermore, the expression level of Sp3 was decreased when Neuro2A cells were demethylated with 5-aza-2'-deoxycytidine, and increasing Sp3 levels in Schneider's Drosophila line 2 cells led to the repression of mDOR promoter activity when the promoter was methylated. These results demonstrate that Sp3 and MBD2 are involved in the transcriptional repression of mDOR in Neuro2A cells through binding to the methylated CpG sites in the promoter region and may play a role in the cell type-specific expression of mDOR. PMID- 12890684 TI - Protein kinase C theta modulates nuclear receptor-corepressor interaction during T cell activation. AB - Transcriptional repression by nuclear receptor corepressors plays a critical role in T cell development. However, the role of these corepressors in T cell activation is poorly understood. We report that T cell activation silenced transcription driven by nuclear receptors retinoic acid receptor, retinoid X receptor, and thyroid hormone receptor and induced silencing mediator of retinoic acid and thyroid hormone receptors (SMRT)-receptor interaction. Whereas the expression of a dominant active mutant of protein kinase C theta(PKC theta) induced strong SMRT-receptor interaction in the absence of T cell activation, a dominant negative mutant of PKC theta decreased the interaction. Loss of PKC theta expression by induction of "RNA interference" resulted in the attenuation of basal and activation-induced SMRT-receptor interaction. We suggest that T cell activation silences nuclear receptor-dependent transactivation in part through PKC theta-dependent enhancement of SMRT-receptor interaction. PMID- 12890685 TI - Xeroderma pigmentosum group C protein possesses a high affinity binding site to human centrin 2 and calmodulin. AB - Human centrin 2 (HsCen2), a member of the EF-hand superfamily of Ca2+-binding proteins, is commonly associated with centrosome-related structures. The protein is organized in two domains, each containing two EF-hand motifs, but only the C terminal half exhibits Ca2+ sensor properties. A significant fraction of HsCen2 is localized in the nucleus, where it was recently found associated with the xeroderma pigmentosum group C protein (XPC), a component of the nuclear excision repair pathway. Analysis of the XPC sequence (940 residues), using a calmodulin target recognition software, enabled us to predict two putative binding sites. The binding properties of the two corresponding peptides were investigated by isothermal titration calorimetry. Only one of the peptides (P1-XPC) interacts strongly (Ka = 2.2 x 10(8) m-1, stoichiometry 1:1) with HsCen2 in a Ca2+ dependent manner. This peptide also binds, with a similar affinity (Ka = 1.1 x 10(8) m-1) to a C-terminal construct of HsCen2, indicating that the interaction with the integral protein is mainly the result of the contribution of the C terminal half. The second peptide (P2-XPC) failed to show any detectable binding either to HsCen2 or to its C-terminal lobe. The two peptides interact with different affinities and mechanisms with calmodulin. Circular dichroism and nuclear magnetic resonance were used to structurally characterize the complex formed by the C-terminal domain of HsCen2 with P1-XPC. PMID- 12890686 TI - Transglycosidase activity of chitotriosidase: improved enzymatic assay for the human macrophage chitinase. AB - Chitotriosidase is a chitinase that is massively expressed by lipid-laden tissue macrophages in man. Its enzymatic activity is markedly elevated in serum of patients suffering from lysosomal lipid storage disorders, sarcoidosis, thalassemia, and visceral Leishmaniasis. Monitoring of serum chitotriosidase activity in Gaucher disease patients during progression and therapeutic correction of their disease is useful to obtain insight in changes in body burden on pathological macrophages. However, accurate quantification of chitotriosidase levels by enzyme assay is complicated by apparent substrate inhibition, which prohibits the use of saturating substrate concentrations. We have therefore studied the catalytic features of chitotriosidase in more detail. It is demonstrated that the inhibition of enzyme activity at excess substrate concentration can be fully explained by transglycosylation of substrate molecules. The potential physiological consequences of the ability of chitotriosidase to hydrolyze as well as transglycosylate are discussed. The novel insight in transglycosidase activity of chitotriosidase has led to the design of a new substrate molecule, 4-methylumbelliferyl-(4-deoxy)chitobiose. With this substrate, which is no acceptor for transglycosylation, chitotriosidase shows normal Michaelis-Menten kinetics, resulting in major improvements in sensitivity and reproducibility of enzymatic activity measurements. The novel convenient chitotriosidase enzyme assay should facilitate the accurate monitoring of Gaucher disease patients receiving costly enzyme replacement therapy. PMID- 12890687 TI - The mouse APG10 homologue, an E2-like enzyme for Apg12p conjugation, facilitates MAP-LC3 modification. AB - Autophagy is a process for the bulk degradation of cytosolic compartments by lysosomes/vacuoles. The formation of autophagosomes involves a dynamic rearrangement of the membrane for which two ubiquitin-like modifications (the conjugation of Apg12p and the modification of a soluble form of MAP-LC3 to a membrane-bound form) are essential. In yeast, Apg10p is an E2-like enzyme essential for Apg12p conjugation. The isolated mouse APG10 gene product interacts with mammalian Apg12p dependent on mammalian Apg7p (E1-like enzyme), and facilitates Apg12p conjugation. The interaction of Apg10p with Apg12p is dependent on the carboxyl-terminal glycine of Apg12p. Mutational analysis of the predicted active site cysteine (Cys161) within mouse Apg10p shows that mutant Apg10pC161S, which can form a stable intermediate with Apg12p, inhibits Apg12p conjugation even in the presence of Apg7p, while overexpression of Apg7p facilitates formation of an Apg12p-Apg5p conjugate. Furthermore, the coexpression of Apg10p with Apg7p facilitates the modification of a soluble form of MAP-LC3 to a membrane-bound form, a second modification essential for autophagy. Mouse Apg10p interacts with MAP-LC3 in HEK293 cells, while no mutant Apg10pC161S forms any intermediate with MAP-LC3. Direct interaction between Apg10p and MAP-LC3 is also demonstrated by yeast two-hybrid analysis. The inability of mutant Apg10pC161S to form any intermediate with MAP-LC3 has ruled out the possibility that MAP-LC3 interacts with Apg10p as a substrate. PMID- 12890688 TI - The BRCA1/BARD1 heterodimer assembles polyubiquitin chains through an unconventional linkage involving lysine residue K6 of ubiquitin. AB - The BRCA1 tumor suppressor forms a heterodimer with the BARD1 protein, and the resulting complex functions as an E3 ubiquitin ligase that catalyzes the synthesis of polyubiquitin chains. In theory, polyubiquitination can occur by isopeptide bond formation at any of the seven lysine residues of ubiquitin. The isopeptide linkage of a polyubiquitin chain is a particularly important determinant of its cellular function, such that K48-linked chains commonly target proteins for proteasomal degradation, while K63 chains serve non-proteolytic roles in various signaling pathways. To determine the isopeptide linkage formed by BRCA1/BARD1-dependent polyubiquitination, we purified a full-length heterodimeric complex and compared its linkage specificity with that of E6-AP, an E3 ligase known to induce proteolysis of its cellular substrates. Using a comprehensive mutation analysis, we found that E6-AP catalyzes the synthesis of K48-linked polyubiquitin chains. In contrast, however, the BRCA1/BARD1 heterodimer directs polymerization of ubiquitin primarily through an unconventional linkage involving lysine residue K6. Although heterologous substrates of BRCA1/BARD1 are not known, BRCA1 autoubiquitination occurs principally by conjugation with K6-linked polymers. The ability of BRCA1/BARD1 to form K6-linked polyubiquitin chains suggests that it may impart unique cellular properties to its natural enzymatic substrates. PMID- 12890690 TI - Nicotinamide: a potential addition to the anti-psoriatic weaponry. AB - Psoriasis is an inflammatory disorder characterized by a T helper type 1 cell cytokine pattern. Increased expression of adhesion molecules, prominent neutrophil accumulation, and increased production of nitric oxide are characteristics of this disorder. Moreover, histamine and proteases are supposed to participate in the pathogenesis of psoriasis. Nicotinamide is an inhibitor of poly (ADP-ribose) polymerase-1 (PARP-1) that, through enhancement of nuclear kappa B-mediated transcription, plays a pivotal role in the expression of inflammatory cytokines, chemokines, adhesion molecules, and inflammatory mediators. Through interaction with CD38 and inhibition of IL-1, IL-12, and TNF alpha production, nicotinamide produces a mild TH2 bias. Nicotinamide is a potent phosphodiesterase inhibitor and suppresses neutrophil chemotaxis and mast cell histamine release. It inhibits nitric oxide synthase mRNA induction and suppresses antigen-induced lymphocyte transformation. Nicotinamide increases the biosynthesis of ceramides, which upon degradation produce sphingosine. Sphingosine inhibits protein kinase C (PKC) and decreases basal cell proliferation dependent on PKC. Taken together, it can be reasoned that nicotinamide could be a useful addition to anti-psoriatic armamentarium. The combination of nicotinamide and thalidomide or methotrexate provided a powerful synergistic inhibition of murine collagen-induced arthritis. Nicotinamide decreased the methotrexate-induced hepatotoxicity. The above combinations may prove to have a powerful anti-psoriatic effect as well. As PARP inhibitors could exert anti-retroviral effect, nicotinamide could also be of special value in the treatment of HIV-infected psoriatics. PMID- 12890691 TI - Cyclic AMP compartmentation due to increased cAMP-phosphodiesterase activity in transgenic mice with a cardiac-directed expression of the human adenylyl cyclase type 8 (AC8). AB - Hearts from AC8TG mice develop a higher contractility (LVSP) and larger Ca2+ transients than NTG mice, with (surprisingly) no modification in L-type Ca2+ channel current (ICa,L) (1). In this study, we examined the cardiac response of AC8TG mice to beta-adrenergic and muscarinic agonists and IBMX, a cyclic nucleotide phosphodiesterase (PDE) inhibitor. Stimulation of LVSP and ICa,L by isoprenaline (ISO, 100 nM) was twofold smaller in AC8TG vs. NTG mice. In contrast, IBMX (100 microM) produced a twofold higher stimulation of ICa,L in AC8TG vs. NTG mice. IBMX (10 microM) increased LVSP by 40% in both types of mice, but contraction and relaxation were hastened in AC8TG mice only. Carbachol (10 microM) had no effect on basal contractility in NTG hearts but decreased LVSP by 50% in AC8TG mice. PDE assays demonstrated an increase in cAMP-PDE activity in AC8TG hearts, mainly due to an increase in the hydrolytic activity of PDE4 and PDE1 toward cAMP and a decrease in the activity of PDE1 and PDE2 toward cGMP. We conclude that cardiac expression of AC8 is accompanied by a rearrangement of PDE isoforms, leading to a strong compartmentation of the cAMP signal that shields L type Ca2+ channels and protects the cardiomyocytes from Ca2+ overload. PMID- 12890692 TI - Hypothalamic levels of NPY, MCH, and prepro-orexin mRNA during pregnancy and lactation in the rat: role of prolactin. AB - Pregnancy and lactation provide excellent models of physiological hyperphagia and hyperprolactinemia. To identify possible factors associated with the increased feeding in these situations, we measured hypothalamic mRNA levels of three orexigenic neuropeptides--NPY, MCH, and orexins--in nonpregnant, pregnant, and lactating rats by in situ hybridization. NPY mRNA content in the arcuate nucleus was significantly increased during pregnancy and lactation. However, MCH and prepro-orexin expression was decreased in both states. 48 or 72 h of fasting in pregnant and lactating rats further elevated NPY mRNA levels and increased the low MCH mRNA content. Surprisingly, no effect was observed in prepro-orexin mRNA levels. Finally, we investigated the possible effect of high PRL levels on these orexigenic signals using a model of hyperprolactinemia induced by pituitary graft. NPY mRNA content was unchanged, but MCH and prepro-orexin mRNA levels were significantly decreased. Our results suggest that the increased NPY expression might be partly responsible for the hyperphagia observed during pregnancy and lactation. MCH and prepro-orexin may be involved in the adaptation of other homeostatic mechanisms and their decreased levels in these physiological settings could be mediated by the elevated circulating PRL levels. PMID- 12890693 TI - Release and interconversion of P2 receptor agonists by human osteoblast-like cells. AB - Nucleotides, acting as agonists at P2 receptors, are important extracellular signaling molecules in many tissues. In bone they affect both bone-forming osteoblast and bone-resorbing osteoclast cell activity. The presence of nucleotides in the extracellular microenvironment is largely determined by their release from cells and metabolism by ecto-enzymes, both of which have scarcely been studied in bone. We have investigated adenosine 5'-triphosphate (ATP) release from SaOS-2 osteoblastic cells and the activities of cell surface ecto enzymes on ATP metabolism. ATP, but not LDH, was detected in SaOS-2 cell conditioned medium, suggesting these cells were actively releasing ATP. Introduction of ADP resulted in increased ATP concentrations in the medium, which was found not to be receptor mediated. Nucleotide inhibition and substrate specificity studies revealed an ecto-nucleoside diphosphokinase (ecto-NDPK) was responsible for the ADP-->ATP conversion; PCR and immunocytochemistry confirmed its presence. Analysis of ATP metabolism over time demonstrated overall ATP degradation was increased by inhibiting ecto-NDPK activity; confirming that the combined action of multiple osteoblast-expressed ecto-enzymes affected extracellular nucleotide concentration. The data establish the coexistence of ATP consuming, and for the first time, ATP-generating activities on the osteoblast cell surface, the discovery of which has significant implications for studies involving P2 receptor subtypes in bone. PMID- 12890694 TI - The sphingosine kinase 1/sphingosine-1-phosphate pathway mediates COX-2 induction and PGE2 production in response to TNF-alpha. AB - In this study we addressed the role of sphingolipid metabolism in the inflammatory response. In a L929 fibroblast model, tumor necrosis factor-alpha (TNF) induced prostaglandin E2 (PGE2) production by 4 h and cyclooxygenase-2 (COX 2) induction as early as 2 h. This TNF-induced PGE2 production was inhibited by NS398, a COX-2 selective inhibitor. GC-MS analysis revealed that only COX-2 generated prostanoids were produced in response to TNF, thus providing further evidence of COX-2 selectivity. As sphingolipids have been implicated in mediating several actions of TNF, their role in COX-2 induction and PGE2 production was evaluated. Sphingosine-1-phosphate (S1P) induced both COX-2 and PGE2 in a dose responsive manner with an apparent ED50 of 100-300 nM. The related sphingolipid sphingosine also induced PGE2, though with much less efficacy. TNF induced a 3.5 fold increase in sphingosine-1-phosphate levels at 10 min that rapidly returned to baseline by 40 min. Small interfering RNAs (siRNAs) directed against mouse SK1 decreased (typically by 80%) SK1 protein and inhibited TNF-induced SK activity. Treatment of cells with RNAi to SK1 but not SK2 almost completely abolished the ability of TNF to induce COX-2 or generate PGE2. By contrast, cells treated with RNAi to S1P lyase or S1P phosphatase enhanced COX-2 induction leading to enhanced generation of PGE2. Treatment with SK1 RNAi also abolished the effects of exogenous sphingosine and ceramide on PGE2, revealing that the action of sphingosine and ceramide are due to intracellular metabolism into S1P. Collectively, these results provide novel evidence that SK1 and S1P are necessary for TNF to induce COX-2 and PGE2 production. Based on these findings, this study indicates that SK1 and S1P could be implicated in pathological inflammatory disorders and cancer. PMID- 12890695 TI - Bystander effect induced by counted high-LET particles in confluent human fibroblasts: a mechanistic study. AB - The possible mechanism of a radiation-induced bystander response was investigated by using a high-LET heavy particle microbeam, which allows selected cells to be individually hit with precise numbered particles. Even when only a single cell within the confluent culture was hit by one particle of 40Ar (approximately 1260 keV/microm) or 20Ne (approximately 380 keV/microm), a 1.4-fold increase of micronuclei (MN) was detected demonstrating a bystander response. When the number of targeted cells increased, the number of MN biphasically increased; however, the efficiency of MN induction per targeted cell markedly decreased. When 49 cells in the culture were individually hit by 1 to 4 particles, the production of MN in the irradiated cultures were approximately 2-fold higher than control levels but independent of the number and LET of the particles. MN induction in the irradiated-culture was partly reduced by treatment with DMSO, a scavenger of reactive oxygen species (ROS), and was almost fully suppressed by the mixture of DMSO and PMA, an inhibitor of gap junctional intercellular communication (GJIC). Accordingly, both ROS and GJIC contribute to the above-mentioned bystander response and GJIC may play an essential role by mediating the release of soluble biochemical factors from targeted cells. PMID- 12890696 TI - Differential regulation of steroid 5alpha-reductase isozymes expression by androgens in the adult rat brain. AB - The enzyme 5alpha-reductase (5alpha-R) is present in many mammalian tissues, including the brain. The physiological importance of 5alpha-R in the brain derives from its capability to convert testosterone (T) to a more potent androgen, dihydrotestosterone (DHT), and to convert progesterone and deoxycorticosterone (DOC) to their respective 5alpha-reduced derivatives, precursors of allopregnanolone and tetrahydroDOC, potent allosteric modulators of the gamma-aminobutyric acid receptor (GABA(A)-R). 5alpha-R occurs as two isoforms, 5alpha-R type 1 (5alpha-R1) and 5alpha-R type 2 (5alpha-R2). We studied the effects of T and DHT on the mRNA levels of both 5alpha-R isozymes in the prefrontal cortex of the adult rat, using an accurate and precise method that combines the high specificity of one-step quantitative RT-PCR with the sensitivity of capillary electrophoresis. Our results demonstrate that both isozymes of 5alpha-R are expressed in the cerebral cortex of adult rats. The gene expression of 5alpha-R type 2 is under the positive control of T and DHT. The gene that codes for 5alpha-R type 1 is not constitutive, because its expression is negatively regulated by T and DHT. These results open up a new research line that may lead to a better understanding of the role of 5alpha-R isozymes in the physiology of the central nervous system. PMID- 12890697 TI - A novel cellular marker of insulin resistance and early atherosclerosis in humans is related to impaired fat cell differentiation and low adiponectin. AB - The epidemic increase in type 2 diabetes can be prevented only if markers of risk can be identified and used for early intervention. We examined the clinical phenotype of individuals characterized by normal or low IRS-1 protein expression in fat cells as well as the potential molecular mechanisms related to the adipose tissue. Twenty-five non-obese individuals with low or normal IRS-1 expression in subcutaneous abdominal fat cells were extensively characterized and the results compared with 71 carefully matched subjects with or without a known genetic predisposition for type 2 diabetes. In contrast to the commonly used risk marker, known heredity for diabetes, low cellular IRS-1 identified individuals who were markedly insulin resistant, had high proinsulin and insulin levels, and exhibited evidence of early atherosclerosis measured as increased intima media thickness in the carotid artery bulb. Circulating levels of adiponectin were also significantly reduced. Gene analyses of fat cells in a parallel study showed attenuated expression of several genes related to fat cell differentiation (adiponectin, aP2, PPARgamma, and lipoprotein lipase) in the group of individuals characterized by a low IRS-1 expression and insulin resistance. A low IRS-1 expression in fat cells is a marker of insulin resistance and risk for type 2 diabetes and is associated with evidence of early vascular complications. Impaired adipocyte differentiation, including low gene expression and circulating levels of adiponectin, can provide a link between the cellular marker and the in vivo phenotype. PMID- 12890698 TI - HIP/PAP stimulates liver regeneration after partial hepatectomy and combines mitogenic and anti-apoptotic functions through the PKA signaling pathway. AB - The HIP/PAP (=human Reg-2) C-type lectin encoding gene is activated in primary liver cancers. In normal liver, the protein is undetectable in normal mature hepatocytes and found only in some ductular cells, representing potential hepatic progenitor cells. The aim of this study was to examine the consequences of human HIP/PAP expression in the liver of transgenic mice. We demonstrated that HIP/PAP stimulated liver regeneration after partial hepatectomy. To further investigate the enhanced liver regeneration observed in vivo, primary cultures of hepatocytes were used to evaluate the mitogenic and anti-apoptotic properties of HIP/PAP. HIP/PAP increased hepatocyte DNA synthesis and protected hepatocytes against TNF alpha plus actinomycin-D-induced apoptosis. HIP/PAP anti-apoptotic effects against TNF-alpha were clearly demonstrated when protein kinase A activity was specifically inhibited by KT5720, and HIP/PAP stimulated PKA-dependent phosphorylation of the proapoptotic Bad protein at Ser-112, suggesting that HIP/PAP may compete with cAMP to stimulate PKA activity. Overall, our results led us to propose a new role for a C-type lectin, HIP/PAP, as a hepatic cytokine that combines mitogenic and anti-apoptotic functions regarding hepatocytes, and consequently acts as a growth factor in vivo to enhance liver regeneration. PMID- 12890699 TI - Alterations in the vascular extracellular matrix of granulocyte macrophage colony stimulating factor (GM-CSF) -deficient mice. AB - GM-CSF takes part in the cytokine network regulating the metabolism of extracellular matrix (ECM) during atherogenesis. Since data also point to an effect of GM-CSF on the vascular ECM in general, the vascular collagenous matrix was studied in wild-type and GM-CSF-deficient mice. Histological examination revealed a disorganized vascular ECM in GM-CSF-deficient mice involving the collagenous matrix and elastic fiber system. As shown by electron microscopy, collagen bundles were disrupted and reduced. The diameter of fibrils varied widely. mRNA expression of collagens and related molecules was studied. Fibrillar collagens were markedly reduced, alpha1(I)procollagen to 16.5% of control levels alpha1(III)procollagen was abolished whereas the expression level of network forming alpha1(VIII)procollagen was not altered. As shown by in situ hybridization, the number of collagen-expressing cells was reduced. Matrix metalloproteinases and their inhibitor 1 were not affected by GM-CSF deficiency. Our studies demonstrate that GM-CSF plays a major role in the cytokine network regulating the metabolism of vascular collagens. GM-CSF deficiency leads to an altered composition of the vascular collagenous matrix, i.e., reduced amount of fibrillar collagen, altered ratio of fibrillar and network-forming collagen, and failures in the fibrillogenesis. We suggest that GM-CSF is a basic requirement for the maintenance of vessel wall integrity and resilience. PMID- 12890700 TI - Platelet endothelial cell adhesion molecule-1 modulates endothelial cell motility through the small G-protein Rho. AB - Platelet endothelial cell adhesion molecule-1 (PECAM-1), an immunoglobulin family vascular adhesion molecule, is involved in endothelial cell migration and angiogenesis (1, 2). We found that endothelial cells lacking PECAM-1 exhibit increased single cell motility and extension formation but poor wound healing migration, reminiscent of cells in which Rho activity has been suppressed by overexpressing a GTPase-activating protein (3). The ability of PECAM-1 to restore wound healing migration to PECAM-1-deficient cells was independent of its extracellular domain or signaling via its immunoreceptor tyrosine-based inhibitory motif. PECAM-1-deficient endothelial cells had a selective defect in RhoGTP loading, and inhibition of Rho activity mimicked the PECAM-1-deficient phenotype of increased chemokinetic single cell motility at the expense of coordinated wound healing migration. The wound healing advantage of PECAM-1 positive endothelial cells was not only Rho mediated but pertussis toxin inhibitable, characteristic of migration mediated by heterotrimeric G-protein linked seven-transmembrane receptor signaling such as signaling in response to the serum sphingolipid sphingosine-1-phosphate (S1P) (4, 5). Indeed, we found that the wound healing defect of PECAM-1 null endothelial cells is minimized in sphingolipid-depleted media; moreover, PECAM-1 null endothelial cells fail to increase their migration in response to S1P. We have also found that PECAM-1 localizes to rafts and that in its absence heterotrimeric G-protein components are differentially recruited to rafts, providing a potential mechanism for PECAM 1-mediated coordination of S1P signaling. PECAM-1 may thus support the effective S1P/RhoGTP signaling required for wound healing endothelial migration by allowing for the spatially directed, coordinated activation of Galpha signaling pathways. PMID- 12890701 TI - Does metabolic radiolabeling stimulate the stress response? Gene expression profiling reveals differential cellular responses to internal beta vs. external gamma radiation. AB - DNA microarray analyses were used to investigate the effect of cell-incorporated 35S-methionine on human colorectal carcinoma cells. This beta-radiation-induced gene expression profile was compared with that induced by external gamma radiation. The extent of DNA fragmentation was used as a biomarker to determine the external gamma dose that was bioequivalent to that received by cells incubated in medium containing 35S-methionine. Studies showed that 35S-methionine at 100 microCi/mL induced a much more robust transcriptional response than gamma radiation (2000 cGy) when evaluated 2 h after the labeling or irradiation period. The cellular response to internal beta-radiation was greater not only with respect to the number of genes induced, but also with respect to the level of gene induction. Not surprisingly, the induced genes overlapped with the set of gamma-responsive genes. However, a distinct beta-gene induction profile that included a large number of cell adhesion proteins was also observed. Taken together, these studies demonstrate that metabolic incorporation of a low energy beta-emitter, such as 35S-methionine, can globally influence a diverse set of cellular activities that can, in turn, affect the outcome of many experiments by altering the cell cycle, metabolic, signaling, or redox status (set point) of the cell. Additional studies of the mechanism of beta-induced proliferation arrest and cell death and of the significance of its differential gene induction/repression profile in comparison to pulsed gamma-irradiation may lead to new insights into the ways in which ionizing radiation can interact with cells. PMID- 12890702 TI - Increased peripheral resistance in heart failure: new evidence suggests an alteration in vascular smooth muscle function. AB - Increased peripheral resistance is a hallmark of chronic heart failure and has been primarily attributed to neurohumoral pathways involving both the renin angiotensin and sympathetic nervous systems. The increased resistance is thought to serve as a compensatory mechanism to help maintain perfusion to the vital organs by sustaining blood pressure in the fate of a failing heart. Local mechanisms, and in particular endothelial dysfunction, have also been shown to be important contributors in regulating arterial resistance and vascular remodeling in this disease. In this issue of the British Journal of Pharmacology, Gschwend et al. (2003) present new data suggesting that in the absence of a functional endothelium, myogenic constriction of small pressurized mesenteric arteries, an intrinsic property of vascular smooth muscle cells, is enhanced in a coronary artery ligation-induced myocardial infarction model of congestive heart failure (CHF) in the rat. The increased myogenic tone appears to be tightly linked to angiotensin II type 1 receptors (AT(1)). The possibility that CHF-induced stimulation of myogenic constriction is due to the local release of preformed angiotensin II or constitutive upregulation of the AT(1) receptor signaling pathways are discussed along with other potential cellular and molecular mechanisms previously suggested to play a role in myogenic reactivity. PMID- 12890703 TI - Acute effects of oestrogen receptor subtype-specific agonists on vascular contractility. AB - This study shows for the first time that both the putatively selective oestrogen receptor alpha and oestrogen receptor beta agonists PPT (4,4',4"-(4-propyl-[(1)H] pyrazole-1,3,5-triyl) tris-phenol) and DPN (2,3-bis(4-hydroxyphenyl) propionitrile) can acutely relax precontracted isolated rat mesenteric arteries at pharmacological (i.e. micro M) concentrations. When compared to responses observed to similar concentrations of 17beta-oestrogen obtained on the same tissues, PPT had a significantly greater vasodilatory effect, while DPN had a significantly smaller effect. All responses were rapid being complete within 5 min exposure time. Thus, both PPT and DPN can acutely relax isolated mesenteric arteries with the relative potency of PPT>17beta-oestrogen>DPN. PMID- 12890704 TI - Structural requisites of 2-(p-chlorophenoxy)propionic acid analogues for activity on native rat skeletal muscle chloride conductance and on heterologously expressed CLC-1. AB - (1) The 2-(p-chlorophenoxy)propionic acid (CPP) modulates in a stereoselective manner the macroscopic chloride conductance (gCl), the electrical parameter sustained by the CLC-1 channel, of skeletal muscle. In order to determine the structural requirements for modulating native gCl and to identify high-affinity ligands, the effects of newly synthesised CPP analogues have been evaluated on gCl of rat EDL muscle fibres by means of the two-microelectrode current-clamp technique. (2) Each type of the following independent modification of CPP structure led to a three- to 10-fold decrease or to a complete lack of gCl blocking activity: replacement of the electron-attractive chlorine atom of the aromatic ring, substitution of the oxygen atom of the phenoxy group, modification at the chiral centre and substitution of the carboxylic function with a phosphonate one. (3) The analogues bearing a second chlorophenoxy group on the asymmetric carbon atom showed a significant gCl-blocking activity. Similar to racemate CPP, the analogue with this group, spaced by an alkyl chain formed by three methylenic groups, blocked gCl by 45% at 100 micro M. (4) These latter derivatives were tested on heterelogously expressed CLC-1 performing inside-out patch-clamp recordings to further define how interaction between drug and channel protein could take place. Depending on the exact chemical nature of modification, these derivatives strongly blocked CLC-1 with K(D) values at -140 mV ranging from about 4 to 180 micro M. (5) In conclusion, we identified four molecular determinants pivotal for the interaction with the binding site on muscle CLC-1 channels: (a) the carboxylic group that confers the optimal acidity and the negative charge; (b) the chlorophenoxy moiety that might interact with a hydrophobic pocket; (c) the chiral centre that allows the proper spatial disposition of the molecule; (d) an additional phenoxy group that remarkably stabilises the binding by interacting with a second hydrophobic pocket. PMID- 12890705 TI - Diadenosine-5-phosphate exerts A1-receptor-mediated proarrhythmic effects in rabbit atrial myocardium. AB - (1) Diadenosine polyphosphates have been described to be present in the myocardium and exert purinergic- and nonreceptor-mediated effects. Since the electrophysiological properties of atrial myocardium are effectively regulated by A(1) receptors, we investigated the effect of diadenosine pentaphosphate (Ap(5)A) in rabbit myocardium. (2) Parameters of supraventricular electrophysiology and atrial vulnerability were measured in Langendorff-perfused rabbit hearts. Muscarinic potassium current (I(K(ACh/Ado))) and ATP-sensitive potassium current (I(K(ATP))) were measured by using the whole-cell voltage clamp method. (3) Ap(5)A prolonged the cycle length of spontaneously beating Langendorff perfused hearts from 225+/-14 (control) to 1823+/-400 ms (Ap(5)A 50 micro M; n=6; P<0.05). This effect was paralleled by higher degree of atrio-ventricular block. Atrial effective refractory period (AERP) in control hearts was 84+/-14 ms (n=6). Ap(5)A>/=1 micro M reduced AERP (100 micro M, 58+/-11 ms; n=6). (4) Extrastimuli delivered to hearts perfused with Ap(5)A- or adenosine (>/= micro M)-induced atrial fibrillation, the incidence of which correlated to the concentration added to the perfusate. The selective A(1)-receptor antagonist CPX (20 micro M) inhibited the Ap(5)A- and adenosine-induced decrease of AERP. Atrial fibrillation was no longer observed in the presence of CPX. (5) The described Ap(5)A-induced effects in the multicellular preparation were enhanced by dipyridamole (10 micro M), which is a cellular adenosine uptake inhibitor. Dipyridamole-induced enhancement was inhibited by CPX. (6) Ap(5)A (5 mm diameter) (P < 0.001) and was not correlated with healthy status. To assess the biological activity of ovarian PPARgamma, ovine granulosa cells were transfected with a reporter construct driven by PPARgamma-responsive elements. Addition of rosiglitazone, a PPARgamma ligand, stimulated reporter gene expression, showing that endogenous PPARgamma is functional in ovine granulosa cells in vitro. Moreover, rosiglitazone inhibited granulosa cell proliferation (P < 0.05) and increased the secretion of progesterone in vitro (P < 0.05). This stimulation effect was stronger in granulosa cells from small than from large follicles. In contrast, rosiglitazone had no effect on LH, FSH, prolactin and growth hormone secretion by ovine pituitary cells in vitro. Overall, these data suggest that PPARgamma ligands might stimulate follicular differentiation in vivo likely through a direct action on granulosa cells rather than by modulating pituitary hormone secretion. PMID- 12890737 TI - Cell cycle duration at the time of maternal zygotic transition for in vitro produced bovine embryos: effect of oxygen tension and transcription inhibition. AB - Early embryonic cleavages are mostly regulated by maternal components then control of development progressively depends on newly synthesized zygotic products. The timing of the first cleavages is a way to assess embryo quality. The goal of this study was to evaluate the duration of the fourth cell cycle, at the time of maternal-to-zygotic transition (MZT) in in vitro-produced bovine embryos by means of cinematographic analysis. We found that 75% of the embryos displayed a long fourth cycle (43.5 +/- 5.4 h) whereas the remaining embryos had a very short fourth cell cycle (8.9 +/- 2.9 h). Both groups did not differ in cleavage rhythm up to the eight-cell stage and timing of cavitation and blastocyst expansion was identical. However, embryos with a short fourth cell cycle had a better blastocyst rate than embryos with a long cycle (59% versus 38%, P < 0.01). Total cell number, inner cell mass (ICM):total cell ratio, and hatching rate were identical for blastocysts produced from embryos with either a long or a short fourth cell cycle. In a second experiment, we showed that increasing the oxygen tension, from 5% to 20%, decreased the percentage of embryos with a short fourth cell cycle, from 25% to 11% (P < 0.01), indicating that suboptimal culture conditions can influence the length of this cycle. Finally, we investigated whether fourth cell cycle duration could be influenced by transcription inhibition. With alpha-amanitin added at 18 h postinsemination (HPI), cleavage was reduced (66% versus 79%) and, at 70 HPI, the 9- to 16-cell rate increased (50% versus 25%) concomitantly with a 5- to 8-cell rate decrease (16% versus 47%). A similar pattern was observed when the drug was added at 6 HPI or 42 HPI but not at 0 HPI. Cinematographic analysis revealed that alpha-amanitin increased the first cell cycle duration whereas the second and third cell cycles were not affected. With the drug, one third of the embryos could develop up to the 9- to 16-cell stage and they all had a short fourth cell cycle (11.2 +/- 3.7 h) with a good synchrony of cleavage between blastomeres. These results suggest that duration of the fourth cell cycle of bovine embryo, during the MZT, is under a zygotic transcriptional control that can be affected by oxidative conditions. PMID- 12890738 TI - Normospermic versus teratospermic domestic cat sperm chromatin integrity evaluated by flow cytometry and intracytoplasmic sperm injection. AB - Teratospermia (>60% of morphologically abnormal spermatozoa) is well documented in felids. Even morphologically normal spermatozoa from teratospermic ejaculates have reduced ability to undergo tyrosine phosphorylation, acrosome react, and bind and penetrate oocytes compared with normospermic (<40% abnormal spermatozoa) counterparts. However, it is unknown whether fertilization deficiencies originate at a nuclear level. This study examined whether fertilization failure also was attributable to abnormal sperm chromatin, using the sperm chromatin structure assay (SCSA), in vitro fertilization (IVF), and intracytoplasmic sperm injection (ICSI). Aliquots of unprocessed and swim-up-processed (to isolate morphologically normal spermatozoa) spermatozoa from teratospermic and normospermic domestic cats were analyzed by the flow cytometric SCSA. Swim-up-processed sperm were incubated with in vivo-matured oocytes or used for ICSI. Teratospermic ejaculates expressed more (P < 0.05) chromatin heterogeneity (abnormal chromatin structure) than their normospermic counterparts, both in unprocessed and swim-up-processed samples. Fertilization success in vitro was higher (P < 0.05) from normo- compared with teratospermic inseminates. Similar (P > 0.05) proportions of oocytes fertilized after ICSI using spermatozoa from normo- and teratospermic cats. Results reveal that teratospermia in the cat is expressed at the nuclear level as increased sperm chromatin heterogeneity, but ICSI showed that this does not apparently affect fertilization rates if the zona pellucida and oolemma can be bypassed. PMID- 12890739 TI - Emerging roles of BRCA1 alternative splicing. AB - Germline mutations of the BRCA1 gene predispose individuals mainly to the development of breast and/or ovarian cancer. However, the exact function of the gene is still unclear, although the encoded proteins are involved in various cellular processes, including transcriptional regulation and DNA repair pathways. Several BRCA1 splice variants are found in different tissues, but in spite of intense investigations, their regulation and possible functions are poorly understood at the moment. This review summarises current knowledge on the roles of these splice variants and the mechanisms responsible for their formation. Because alternative splicing is now widely accepted as an important source of genetic diversity, elucidating the functions of the BRCA1 splice variants would help in the understanding of the exact role(s) of this tumour suppressor. This should help to resolve the current paradox that, despite its seemingly vital cellular functions, mutations of this gene are associated with tissue specific tumour formation predominantly in the breast and the ovary. PMID- 12890740 TI - Demystified...tissue microarray technology. AB - Several "high throughput methods" have been introduced into research and routine laboratories during the past decade. Providing a new approach to the analysis of genomic alterations and RNA or protein expression patterns, these new techniques generate a plethora of new data in a relatively short time, and promise to deliver clues to the diagnosis and treatment of human cancer. Along with these revolutionary developments, new tools for the interpretation of these large sets of data became necessary and are now widely available. Tissue microarray (TMA) technology is one of these new tools. It is based on the idea of applying miniaturisation and a high throughput approach to the analysis of intact tissues. The potential and the scientific value of TMAs in modern research have been demonstrated in a logarithmically increasing number of studies. The spectrum for additional applications is widening rapidly, and comprises quality control in histotechnology, longterm tissue banking, and the continuing education of pathologists. This review covers the basic technical aspects of TMA production and discusses the current and potential future applications of TMA technology. PMID- 12890741 TI - Molecular detection of the G(-248)A BAX promoter nucleotide change in B cell chronic lymphocytic leukaemia. AB - BACKGROUND: A novel single nucleotide polymorphism (SNP), G(-248)A, in the 5' untranslated region of the BAX promoter and its association with reduced protein expression, progression beyond Rai stage 0, and treatment resistance in chronic lymphocytic leukaemia (CLL) has been reported previously. AIM: To develop a restriction enzyme analysis (REA) based method for routine detection of BAX promoter SNP in a clinical laboratory. METHODS: The BAX promoter was analysed in duplicate by REA and sequencing in 90 samples (from 45 patients with CLL, 43 controls, and two cell lines). The promoter region was amplified, digested with restriction endonucleases (Aci I and Tau I), and separated by gel electrophoresis. RESULTS: After digestion, the normal GG genotype samples produced three distinct bands. The homozygous AA replacement abolished the cleavage site, resulting in a single band. Although the heterozygous samples produced three bands, the two smaller visible bands were reduced in intensity (> 50%). The test characteristics of Aci I REA were better than those of Tau I REA, in terms of sensitivity (100% v 77.8%), specificity (98.6% v 92.3%), positive predictive value (95.03% v 87.4%), and negative predictive value (100% v 85.83%). CONCLUSIONS: REA using Aci I is a highly sensitive and specific method for detecting the BAX G(-248)A SNP in CLL. PMID- 12890742 TI - Expressive genomic hybridisation: gene expression profiling at the cytogenetic level. AB - AIMS: To describe a cytogenetic technique suitable for the rapid assessment of global gene expression that is based on comparative genomic hybridisation (CGH), and to use it to understand the relation between genetic amplifications and gene expression. METHODS: Whereas traditional CGH uses DNA as test and reference in hybridisations, expressive genomic hybridisation (EGH) uses globally amplified mRNA as test and normal DNA as reference. EGH is a rapid and powerful tool for localising and studying global gene expression profiles and correlating them with loci of genetic amplifications using traditional CGH. RESULTS: EGH was used to correlate genetic amplifications detected by CGH with the expression profile of two independent cell lines-Colo320 and T47D. Although many amplifications resulted in overexpression, other amplifications were partially or completely silenced at the cytogenetic level. CONCLUSION: This technique will assist in the analysis of overexpressed genes within amplicons and could resolve a controversial issue in cancer cytogenetics; namely, the relation between genetic amplifications and overexpression. PMID- 12890743 TI - Differential expression of E-cadherin and beta catenin in primary and metastatic Wilms's tumours. AB - BACKGROUND: The E-cadherin-catenin adhesion complex is crucial for intercellular adhesiveness and maintenance of tissue architecture. Its impairment is associated with poorly differentiated phenotype and increased invasiveness of carcinomas. AIMS: To evaluate E-cadherin, beta catenin, gamma catenin, and ezrin expression and its relation to histopathological features of primary and metastatic Wilms's tumours. METHODS: Immunohistochemistry was used to determine the expression and cellular distribution of E-cadherin, beta catenin, gamma catenin, and ezrin in primary and metastatic Wilms's tumours. Western blotting was used to determine polypeptide size and expression of E-cadherin and beta catenin in Wilms's tumours compared with normal kidney. RESULTS: Moderate expression of E-cadherin was found mainly in cytoplasm and occasionally cell membranes of dysplastic tubules, whereas low expression was seen in cytoplasm of blastemal cells. Primary and metastatic tumours showed moderate to high beta catenin expression in blastemal and epithelial cells, with predominantly membranous and cytoplasmic staining. Occasional nuclear staining was noted in metastatic tumours. Low to high gamma catenin and ezrin expression was seen in cytoplasm of blastemal and epithelial cells of primary and metastatic tumours. Higher amounts of 92 kDa beta catenin were detected in tumours than in normal kidney. Low expression of 120 kDa E cadherin was seen in moderately differentiated tumours, whereas expression was lacking in poorly differentiated tumours. CONCLUSIONS: Compared with primary tumours, metastatic tumours showed lower expression of E-cadherin and gamma catenin, with nuclear staining for beta catenin. Low E-cadherin was associated with poorly differentiated tumours. These results suggest that abnormal expression of adhesion proteins correlates with the invasive and metastatic phenotype in Wilms's tumours. PMID- 12890744 TI - Analysis of the tumour suppressor genes, FHIT and WT-1, and the tumour rejection genes, BAGE, GAGE-1/2, HAGE, MAGE-1, and MAGE-3, in benign and malignant neoplasms of the salivary glands. AB - AIMS: Molecular genetic changes involved in tumorigenesis and malignant transformation of human tumours are novel targets of cancer diagnosis and treatment. This study aimed to analyse the expression of putative tumour suppressor genes, FHIT and WT-1, and tumour rejection genes, BAGE, GAGE-1/2, MAGE 1, MAGE-3, and HAGE (which are reported to be important in human cancers), in salivary gland neoplasms. METHODS: Gene expression was analysed by reverse transcription polymerase chain reaction (RT-PCR) in normal salivary gland tissue and 44 benign and malignant salivary gland tumours. RESULTS: Aberrant FHIT transcripts were found in one of 38 normal salivary glands, three of 28 adenomas, and two of 16 carcinomas. WT-1 mRNA was detectable in two adenomas and five carcinomas. Immunoblotting showed that WT-1 mRNA expression was associated with raised WT-1 protein concentrations. RT-PCR for detection of BAGE, GAGE, and MAGE gene expression was positive in two adenomas and nine carcinomas, but negative in normal salivary gland tissue. HAGE mRNA was found in two normal salivary glands, 11 benign, and eight malignant tumours. CONCLUSIONS: FHIT mRNA splicing does not appear to be involved in the genesis of salivary gland neoplasms. The upregulation of WT-1 mRNA in tumours of epithelial/myoepithelial phenotype may imply a potential role of WT-1 in the genesis and/or cellular differentiation of these salivary gland tumours. The tumour rejection genes were more frequently, but not exclusively, expressed in malignant salivary gland tumours than in benign neoplasms, although none was suitable as a diagnostic marker of malignancy in salivary gland neoplasms. PMID- 12890745 TI - Isolation and partial characterisation of a new antiproliferative substance from human leucocytes inhibiting growth of Candida albicans. AB - AIM: To purify and partially characterise a fraction from human leucocytes containing a substance cytotoxic to Candida albicans. METHODS: Leucocytes were isolated from the buffy coats of healthy blood donors. The cytotoxic factor (CF) was isolated from the soluble fraction of the cells. A cell lysate was passed through a filter with a cut off value of 3 kDa, and the filtrate was processed by anionic exchange chromatography and gel filtration. The purified CF was analysed for its chemical and biological properties. The cytotoxicity of CF was tested on C albicans grown on agar plates. RESULTS: Mass spectrometry showed a molecular mass of 2.148 kDa. CF was found in polymorphonuclear neutrophilic cells only. No amino acids were detected, and a low ultraviolet absorbance at 260 nm and resistance to nuclease indicate the absence of nucleic acids. An anthrone test was positive for carbohydrate. The substance was soluble in water. CF showed a dose related cytotoxicity in the range of 0.1-1 mg/ml. The cytotoxic effect was abrogated by zinc ions. Preliminary testing indicated that CF also had cytotoxic effects against some bacteria. CONCLUSIONS: This report describes a factor from isolated human leucocytes that is cytotoxic to C albicans. The substance contains a carbohydrate moiety, whereas no amino acids were detected. The cytotoxicity can be abrogated by zinc ions in vitro. This substance is probably part of the repertoire by which leucocytes prevent infections. PMID- 12890746 TI - Efficiency and cost effectiveness: PAGE-SSCP versus MDE and Phast gels for the identification of unknown beta thalassaemia mutations. AB - BACKGROUND: Prenatal diagnosis for beta thalassaemia has proved to be very effective in preventing the birth of an affected child and hence in controlling the disease. The success of prenatal diagnosis depends on the delineation of the underlying mutations in the population at risk. Each population carries a limited number of frequent defects (89-91%) and a variable number of rare alleles (4-5%), whereas 2-3% of alleles remain uncharacterised. To offer prenatal diagnosis when the parental mutation is unknown, the application of a non-specific detection method (such as single stranded conformational polymorphism (SSCP)) to localise the mutation, followed by direct sequencing of the amplified gene sequence, is required. With this objective in mind, this study was designed to devise the best protocol and system of SSCP for the rapid screening of unknown mutations in the beta globin gene. METHODS: To detect mutations in this disease, three different systems-Phast gels, MDE gels, and polyacrylamide gels-were used under varying conditions. RESULTS: Polyacrylamide gels were found to be the most efficient, both in terms of resolution and cost. CONCLUSION: Polyacrylamide gels are the most rapid, efficient, reliable, and cost effective means for DNA mutation analysis of the beta globin gene. PMID- 12890747 TI - Improved resolution by mounting of tissue sections for laser microdissection. AB - BACKGROUND: Laser microbeam microdissection has greatly facilitated the procurement of specific cell populations from tissue sections. However, the fact that a coverslip is not used means that the morphology of the tissue sections is often poor. AIMS: To develop a mounting method that greatly improves the morphological quality of tissue sections for laser microbeam microdissection purposes so that the identification of target cells can be facilitated. METHODS: Fresh frozen tissue and formalin fixed, paraffin wax embedded tissue specimens were used to test the morphological quality of mounted and unmounted tissue. The mounting solution consisted of an adhesive gum and blue ink diluted in water. Interference of the mounting solution with DNA quality was analysed by the polymerase chain reaction using 10-2000 cells isolated by microdissection from mounted and unmounted tissue. RESULTS: The mounting solution greatly improved the morphology of tissue sections for laser microdissection purposes and had no detrimental effects on the isolation and efficiency of amplification of DNA. One disadvantage was that the mounting solution reduced the cutting efficiency of the ultraviolet laser. To minimise this effect, the mounting solution should be diluted as much as possible. Furthermore, the addition of blue ink to the mounting medium restores the cutting efficiency of the laser. CONCLUSIONS: The mounting solution is easy to prepare and apply and can be combined with various staining methods without compromising the quality of the DNA extracted. PMID- 12890748 TI - Concentrations of circulating matrix metalloproteinase 9 inversely correlate with autoimmune antibodies to double stranded DNA: implications for monitoring disease activity in systemic lupus erythematosus. AB - AIMS: To compare circulating matrix metalloproteinase (MMP) concentrations with antibodies to single and double stranded DNA (ssDNA and dsDNA) to determine their relation in inflammatory arthritic diseases, such as systemic lupus erythematosus (SLE). METHODS: Fibroblast MMP-2 and neutrophil MMP-9 were resolved by gelatin zymography and measured by densitometry. Anti-ssDNA and anti-dsDNA were determined by enzyme immunoassay and samples grouped on antibody content as follows: low anti-ssDNA/low anti-dsDNA antibodies (group 1); high anti-ssDNA/low anti-dsDNA antibodies (group 2); and high anti-ssDNA/high anti-dsDNA antibodies (group 3). RESULTS: Group 3 samples contained significantly lower amounts of MMP 9 when compared with group 1 samples. Higher molecular weight MMP-9 forms (130 and 225 kDa) were virtually absent. Group 2 samples contained intermediate MMP-9 concentrations. Fibroblast MMP-2 was unchanged in all groups. Mean complement C3 and C4 concentrations showed a consistent, but variably significant, decrease with increasing anti-ssDNA and anti-dsDNA antibodies. The mean erythrocyte sedimentation rate was raised in all patient groups. CONCLUSIONS: Neutrophil MMP 9, an inflammatory marker, inversely correlates with anti-dsDNA antibodies, which are a specific marker for SLE, and may be important in monitoring disease activity during antibody deposition in tissues. PMID- 12890749 TI - Does leptin resistance contribute to infections in patients with diabetes? PMID- 12890750 TI - Potential viral pathogenic mechanism for new variant inflammatory bowel disease. PMID- 12890751 TI - Bcl-2 expression decreases cadherin-mediated cell-cell adhesion. AB - Bcl-2, a member of the apoptosis-regulating family of proteins confers a survival advantage on cells by inhibiting apoptosis. Bcl-2 expression is estrogen responsive and high in various tumors. Overexpression of Bcl-2 has been associated with the loss of contact inhibition, unregulated growth and foci formation in culture. In this study, we have examined the effects of bcl-2 overexpression and expression on cell-cell adhesion in MCF-7 and MDCK epithelial cell lines respectively. Overexpression of Bcl-2 in estrogen receptor-positive MCF-7 mammary carcinoma cells led to decreased cell surface E-cadherin and the disruption of junctional complexes concurrent with intracellular redistribution of their components. Particularly noticeable, was the partial nuclear localization of the tight junction-associated protein ZO-1 which coincided with upregulation of ErbB2. The expression of this EGF co-receptor is regulated by the ZO-1-associated transcription factor ZONAB. Growth in estrogen-depleted media led to downregulation of Bcl-2 expression and upregulation and membrane localization of all junctional proteins. Similar disruption in junctions, accompanied by decreased transepithelial resistance, was observed when Bcl-2 was expressed in MDCK cells. These results strongly suggest that Bcl-2 expression decreases the level of functional E-cadherin thereby interfering with junction formation. The inhibition of junction formation decreases cell-cell adhesion leading to the loss of contact inhibition, which, in vivo, can lead to unregulated growth and tumorigenesis. PMID- 12890752 TI - Cross-linking of actin filaments by myosin II is a major contributor to cortical integrity and cell motility in restrictive environments. AB - Cells are frequently required to move in a local environment that physically restricts locomotion, such as during extravasation or metastatic invasion. In order to model these events, we have developed an assay in which vegetative Dictyostelium amoebae undergo chemotaxis under a layer of agarose toward a source of folic acid [Laevsky, G. and Knecht, D. A. (2001). Biotechniques 31, 1140 1149]. As the concentration of agarose is increased from 0.5% to 3% the cells are increasingly inhibited in their ability to move under the agarose. The contribution of myosin II and actin cross-linking proteins to the movement of cells in this restrictive environment has now been examined. Cells lacking myosin II heavy chain (mhcA-) are unable to migrate under agarose overlays of greater than 0.5%, and even at this concentration they move only a short distance from the trough. While attempting to move, the cells become stretched and fragmented due to their inability to retract their uropods. At higher agarose concentrations, the mhcA- cells protrude pseudopods under the agarose, but are unable to pull the cell body underneath. Consistent with a role for myosin II in general cortical stability, GFP-myosin dynamically localizes to the lateral and posterior cortex of cells moving under agarose. Cells lacking the essential light chain of myosin II (mlcE-), have no measurable myosin II motor activity, yet were able to move normally under all agarose concentrations. Mutants lacking either ABP-120 or alpha-actinin were also able to move under agarose at rates similar to wild-type cells. We hypothesize that myosin stabilizes the actin cortex through its cross-linking activity rather than its motor function and this activity is necessary and sufficient for the maintenance of cortical integrity of cells undergoing movement in a restrictive environment. The actin cross-linkers alpha actinin and ABP-120 do not appear to play as major a role as myosin II in providing this cortical integrity. PMID- 12890753 TI - Interphase and monoastral-mitotic phenotypes of overexpressed MAP4 are modulated by free tubulin concentrations. AB - The microtubule-associated protein 4 (MAP4) has recently been shown to counteract destabilization of interphase microtubules caused by catastrophe promotion but not by tubulin sequestering. To address how MAP4 discriminates between destabilization of microtubules by these two mechanisms, we have evaluated the combined phenotypes of MAP4 coexpressed with Op18/stathmin family member derivatives with either catastrophe-promoting or sequestering activities. This approach relies on the finding that overexpression of MAP4 alone stabilizes microtubules during all phases of the cell cycle in human leukemia cells, and causes a potent mitotic block and a dramatic, previously unobserved, phenotype characterized by large monoastral spindles. Coexpression of either catastrophe promoting or tubulin-sequestration-specific Op18 derivatives was found to modulate the activity of ectopic MAP4 during mitosis, but with differential functional outcome. Interestingly, the tubulin-sequestering derivative suppressed the monoastral mitotic phenotype of MAP4 (i.e. coexpression facilitated the formation of functional spindles). To evaluate whether this phenotypic suppression could be explained by tubulin-sequestration-dependent modulation of MAP4 activity, a plasma-membrane-targeted, tubulin-sequestering chimera was constructed to decrease the cytosolic free tubulin concentration substantially. This chimera likewise suppressed the monoastral phenotype caused by overexpression of MAP4, suggesting a direct downregulation of MAP4 activity by reduced free tubulin concentrations. PMID- 12890754 TI - Microtubule and Rac 1-dependent F-actin in growth cones. AB - Extracellular cues control the rate and direction of growth of neuronal processes in large part by regulating the cytoskeleton of the growth cone. The actin filament network of the peripheral region is thought to be the primary target for these cues, with consequences for the advance and organization of microtubules. Binding of laminin to integrin receptors is a cue that accelerates the growth of processes from many types of neurons. It was applied acutely to sympathetic neurons in culture to study its effects on the cytoskeleton of the growth cone. Microtubules advance to the edge of the growth cone and bundle in response to laminin, and it was found that small veils of membrane appear near the ends of some of those microtubules. To examine more clearly the relationship between the microtubules and the appearance of actin-rich structures at the periphery, a low dose of cytochalasin D was used to deplete the peripheral region of the growth cone of pre-existing F-actin. The subsequent addition of laminin resulted in the bundling of ends of dynamic (tyrosinated) microtubules at the distal edge of the growth cone, most of which were associated with foci of F-actin. Observations of labeled actin within living growth cones confirmed that these foci formed in response to laminin. Suppression of microtubule dynamics with drugs eliminated the actin foci; washout of drug restored them. Rac 1 did not co-concentrate with F-actin in the peripheral region of the growth cone in the absence of laminin, but did co-concentrate with the foci of F-actin that formed in response to laminin. Inhibition of Rac 1 functioning prevented the formation of the foci and also inhibited laminin-induced neurite growth with or without cytochalasin. These results indicate that extracellular cues can affect actin in the growth cone via microtubules, as well as affect microtubules via actin. They also point to the mediation of microtubule-dependent accumulation of F-actin at the front of the growth cone as a role of Rac 1 in neurite growth. PMID- 12890755 TI - The C-terminal end of R-Ras contains a focal adhesion targeting signal. AB - R-Ras promotes cell adhesion and activation of integrins through a process that is yet unknown. We show here that active R-Ras (38V) promotes the formation of focal adhesions and a spread cell shape. By contrast, the dominant-negative mutant of R-Ras (43N) reduces the number of focal adhesions, leading to the formation of refractile cells. In adherent cells wild-type R-Ras, activated (38V) R-Ras and endogeous R-Ras were preferentially targeted to focal adhesions, whereas the dominant-negative mutant (43N) of R-Ras was excluded from these structures. Activated mutants of H-Ras and K-Ras were not found in focal adhesions. We dissected R-Ras to find out the determinants that are important for the targeting process. The outermost region in the N-terminus of R-Ras, as well as the intact proline-rich sequence in the C-terminus of RRas that mediates binding to Nck, were not essential. Mutating the potential palmitoylation site (C213A) of RRas results in depalmitoylation and accumulation of R-Ras in the Golgi. Using H-Ras/R-Ras, R-Ras/H-Ras and RRas/K-Ras hybrid molecules we showed that the C-termini (175-218 amino acids) of R-Ras contains the signal for focal adhesions targeting. Exchanging the hypervariable region of H-Ras to R-Ras inhibited the targeting of R-Ras to focal adhesions, whereas H-Ras obtained the ability to localize to focal adhesions after receiving the hypervariable region of R-Ras. This indicates that R-Ras targeting is mediated both by the nucleotide binding status as well as through a specific region in the C-terminus of R-Ras. These results indicate that targeting and activation of R-Ras are linked processes in the formation of focal adhesions. PMID- 12890756 TI - KSHV vFLIP binds to IKK-gamma to activate IKK. AB - When expressed in heterologous cells, the viral FLIP protein (vFLIP) of Kaposi's sarcoma-associated herpesvirus (KSHV) has been reported both to block Fas mediated apoptosis and to activate the NF-kappaB activation pathway by interaction with IkappaB kinase (IKK). In a yeast-two-hybrid screen, we identified IKKgamma as an interacting partner of vFLIP. We expressed fragments of IKKgamma in mammalian cells and bacteria, and identified the central CCR3/4 (amino acids 150-272) as the vFLIP binding region. To investigate the proteins interacting with vFLIP in a KSHV-infected primary effusion lymphoma (PEL) cell line, we immunoprecipitated vFLIP and identified four associated proteins by mass spectrometry: IKK components IKKalpha, beta and gamma, and the chaperone, Hsp90. Using gel filtration chromatography, we demonstrated that a single population of vFLIP in the cytoplasm of PEL cells co-eluted and co-precipitated with an activated IKK complex. An inhibitor of Hsp90, geldanamycin, inhibited IKK's kinase activity induced by vFLIP and killed PEL cells, suggesting that vFLIP activation of IKK contributes to PEL cell survival. PMID- 12890757 TI - Architectural defects in pronuclei of mouse nuclear transplant embryos. AB - Reprogramming somatic nuclear function by transplantation of nuclei into recipient oocytes is associated with a morphological remodeling of the somatic nucleus. Successful cloning of animals by nuclear transplantation (NT) demonstrates that reprogramming somatic cell function is possible. However, low pregnancy rates and high frequencies of lethal abnormalities in animals born suggest that reprogramming is rarely complete. To address this issue, we tested the hypothesis that nuclear transplantation leads to nuclear remodeling deficiencies. We report the identification of several markers of morphological remodeling, or lack thereof, of mouse cumulus cell nuclei after transplantation into oocytes. Notably, nuclear transplant mouse embryos exhibit nuclear assembly of the differentiated cell-specific A-type lamins at the one-cell stage, as a result of misregulation of lamin A gene expression. The transplanted nuclei also display enhanced concentration of the nuclear matrix-associated protein NuMA as a result of translation from maternal mRNA and de novo transcription. The A-kinase anchoring protein 95 (AKAP95), a marker of the nuclear envelope-chromatin interface, is of somatic origin. Furthermore, greater resistance of AKAP95 and DNA to in situ extractions of one-cell stage NT embryos with non-ionic detergent, DNase, RNase and NaCl reflects an enhanced proportion of heterochromatin in these embryos. Passage through first embryonic mitosis does not rescue the defects detected in one-cell stage embryos. We propose that somatic nuclear reprogramming deficiencies by NT might emanate from, at least in part, failure to remodel the somatic nucleus morphologically into a functional embryonic nucleus. PMID- 12890758 TI - Glycogen synthase kinase-3 regulates formation of long lamellipodia in human keratinocytes. AB - During wound healing, keratinocytes initiate migration from the wound edge by extending lamellipodia into a fibronectin-rich provisional matrix. While lamellipodia-like structures are also found in cultured keratinocytes exposed to epidermal growth factor (EGF), the signaling pathway that regulates the formation of these structures is not defined. In cultured human keratinocytes seeded on fibronectin, we found that protein-serine/threonine kinase inhibitors including staurosporine, induced concentration-dependent formation of extended lamellipodia (E-lams). The formation of E-lams was inhibited by the proteintyrosine kinase inhibitors herbimycin A and genistein and augmented by the protein-tyrosine phosphatase inhibitor sodium orthovanadate. Staurosporine treatment induced relocation of tyrosine phosphorylated phospholipase C-gamma1 (PLC-gamma1) to the tips of lamellipodia where actin assembly was initiated. Consistent with an involvement of PLC-gamma1 in E-lam formation, intracellular free calcium (Ca2+) was elevated during the formation of E-lams and conversely, E-lam formation was blocked by intracellular Ca2+ chelation with BAPTA/AM, but not by extracellular reduction of Ca2+ by EGTA. Notably, glycogen synthase kinase-3alpha/beta (GSK 3alpha/beta) was activated by staurosporine as evidenced by reduced phosphorylation on Ser-21/9. Suppression of GSK-3 activity by LiCl2 or by a specific chemical inhibitor, SB-415286, blocked E-lam formation but without altering cell spreading. Furthermore, GSK-3 inhibitors blocked both staurosporine and EGF-induced keratinocyte migration in scratch-wounded cultures. We propose that GSK-3 plays a crucial role in the formation of long lamellipodia in human keratinocytes and is potentially a central regulatory molecule in epithelial cell migration during wound healing. PMID- 12890759 TI - Semaphorin 3F is critical for development of limbic system circuitry and is required in neurons for selective CNS axon guidance events. AB - Little is known about the role of class 3 semaphorins in the development of CNS circuitry. Several class 3 semaphorins, including semaphorin 3F (Sema3F) bind to the receptor neuropilin-2 to confer chemorepulsive responses in vitro. To understand the role of Sema3F in the establishment of neural circuitry in vivo, we have generated sema3F null and sema3F conditional mutant mice. Inspection of the peripheral nervous system in sema3F null mice reveals that Sema3F is essential for the proper organization of specific cranial nerve projections. Analysis of the CNS in sema3F null mice reveals a crucial role for Sema3F in the rostral forebrain, midbrain, and hippocampus in establishing specific Npn-2 (neuropilin-2)-expressing limbic tracts. Furthermore, we identify Sema3F and Npn 2 as the first guidance cue-receptor pair shown to be essential for controlling the development of amygdaloid circuitry. In addition, we provide genetic evidence in vertebrates for a neuronal requirement of a soluble axon guidance cue in CNS axon guidance. Our data reveal a requirement for neuronal Sema3F in the normal development of the anterior commissure in the ventral forebrain and infrapyramidal tract in the hippocampus. Thus, our results show that Sema3F is the principal ligand for Npn-2-mediated axon guidance events in vivo and is a critical determinant of limbic and peripheral nervous system circuitry. PMID- 12890760 TI - Temporal resolution of ensemble visual motion signals in primate retina. AB - Recent studies have examined the temporal precision of spiking in visual system neurons, but less is known about the time scale that is relevant for behaviorally important visual computations. We examined how spatiotemporal patterns of spikes in ensembles of primate retinal ganglion cells convey information about visual motion to the brain. The direction of motion of a bar was estimated by comparing the timing of responses in ensembles of parasol (magnocellular-projecting) retinal ganglion cells recorded simultaneously, using a cross-correlation approach similar to standard models of motion sensing. To identify the temporal resolution of motion signals, spike trains were low-pass filtered before estimating the direction of motion. The filter time constant that resulted in most accurate motion sensing was in the range of 10-50 msec for a range of stimulus speeds and contrasts and approached a lower limit of approximately 10 msec at high speeds and contrasts. This time constant was, on average, comparable to the length of interspike intervals. These findings suggest that cortical neurons could filter their inputs on a time scale of tens of milliseconds, rather than relying on the precise times of individual input spikes, to sense motion most reliably. PMID- 12890761 TI - Brain-derived neurotrophic factor val66met polymorphism affects human memory related hippocampal activity and predicts memory performance. AB - BDNF plays a critical role in activity-dependent neuroplasticity underlying learning and memory in the hippocampus. A frequent single nucleotide polymorphism in the targeting region of the human BDNF gene (val66met) has been associated with abnormal intracellular trafficking and regulated secretion of BDNF in cultured hippocampal neurons transfected with the met allele. In addition, the met allele has been associated with abnormal hippocampal neuronal function as well as impaired episodic memory in human subjects, but a direct effect of BDNF alleles on hippocampal processing of memory has not been demonstrated. We studied the relationship of the BDNF val66met genotype and hippocampal activity during episodic memory processing using blood oxygenation level-dependent functional magnetic resonance imaging and a declarative memory task in healthy individuals. Met carriers exhibited relatively diminished hippocampal engagement in comparison with val homozygotes during both encoding and retrieval processes. Remarkably, the interaction between the BDNF val66met genotype and the hippocampal response during encoding accounted for 25% of the total variation in recognition memory performance. These data implicate a specific genetic mechanism for substantial normal variation in human declarative memory and suggest that the basic effects of BDNF signaling on hippocampal function in experimental animals are important in humans. PMID- 12890762 TI - Rapid critical period induction by tonic inhibition in visual cortex. AB - Mice lacking a synaptic isoform of glutamic acid decarboxylase (GAD65) do not exhibit ocular dominance plasticity unless an appropriate level of GABAergic transmission is restored by direct infusion of benzodiazepines into the brain. To better understand how intracortical inhibition triggers experience-dependent changes, we dissected the precise timing requirement for GABA function in the monocular deprivation (MD) paradigm. Diazepam (DZ) or vehicle solution was infused daily before and/or during 4 d of MD in GAD65 knock-out mice. Extracellular single-unit recordings from the binocular zone of visual cortex were performed at the end of deprivation. We found that a minimum treatment of 2 d near the beginning of MD was sufficient to fully activate plasticity but did not need to overlap the deprivation per se. Extended delay after DZ infusion eventually led to loss of plasticity accompanied by improved intrinsic inhibitory circuit function. Two day DZ treatment just after eye opening similarly closed the critical period prematurely in wild-type mice. Raising wild-type mice in complete darkness from birth delayed the peak sensitivity to MD as in other mammals. Interestingly, 2 d DZ infusion in the dark also closed the critical period, whereas equally brief light exposure during dark-rearing had no such effect. Thus, enhanced tonic signaling through GABA(A) receptors rapidly creates a milieu for plasticity within neocortex capable of triggering a critical period for ocular dominance independent of visual experience itself. PMID- 12890763 TI - Impaired NMDA receptor-mediated postsynaptic function and blunted NMDA receptor dependent persistent pain in mice lacking postsynaptic density-93 protein. AB - Modification of synaptic NMDA receptor (NMDAR) expression influences NMDAR mediated synaptic function and associated persistent pain. NMDARs directly bind to a family of membrane-associated guanylate kinases (MAGUKs) that regulate surface and synaptic NMDAR trafficking in the CNS. We report here that postsynaptic density-93 protein (PSD-93), a postsynaptic neuronal MAGUK, is expressed abundantly in spinal dorsal horn and forebrain, where it colocalizes and interacts with NMDAR subunits NR2A and NR2B. Targeted disruption of the PSD 93 gene reduces not only surface NR2A and NR2B expression but also NMDAR-mediated excitatory postsynaptic currents and potentials, without affecting surface AMPA receptor expression or its synaptic function, in the regions mentioned above. Furthermore, mice lacking PSD-93 exhibit blunted NMDAR-dependent persistent pain induced by peripheral nerve injury or injection of Complete Freund's Adjuvant, although they display intact nociceptive responsiveness to acute pain. PSD-93 appears to be important for NMDAR synaptic targeting and function and to be a potential biochemical target for the treatment of persistent pain. PMID- 12890764 TI - A two-stage unsupervised learning algorithm reproduces multisensory enhancement in a neural network model of the corticotectal system. AB - Multisensory enhancement (MSE) is the augmentation of the response to sensory stimulation of one modality by stimulation of a different modality. It has been described for multisensory neurons in the deep superior colliculus (DSC) of mammals, which function to detect, and direct orienting movements toward, the sources of stimulation (targets). MSE would seem to improve the ability of DSC neurons to detect targets, but many mammalian DSC neurons are unimodal. MSE requires descending input to DSC from certain regions of parietal cortex. Paradoxically, the descending projections necessary for MSE originate from unimodal cortical neurons. MSE, and the puzzling findings associated with it, can be simulated using a model of the corticotectal system. In the model, a network of DSC units receives primary sensory input that can be augmented by modulatory cortical input. Connection weights from primary and modulatory inputs are trained in stages one (Hebb) and two (Hebb-anti-Hebb), respectively, of an unsupervised two-stage algorithm. Two-stage training causes DSC units to extract information concerning simulated targets from their inputs. It also causes the DSC to develop a mixture of unimodal and multisensory units. The percentage of DSC multisensory units is determined by the proportion of cross-modal targets and by primary input ambiguity. Multisensory DSC units develop MSE, which depends on unimodal modulatory connections. Removal of the modulatory influence greatly reduces MSE but has little effect on DSC unit responses to stimuli of a single modality. The correspondence between model and data suggests that two-stage training captures important features of self-organization in the real corticotectal system. PMID- 12890765 TI - Differential properties of astrocyte calcium waves mediated by P2Y1 and P2Y2 receptors. AB - Intercellular spread of Ca2+ waves is the primary manifestation of cell-to-cell communication among astrocytes. Ca2+ waves propagate via the release of a diffusible extracellular messenger that has been identified as ATP. In dorsal spinal astrocytes, Ca2+ waves are mediated by activation of two functionally distinct subtypes of metabotropic purinoceptor: the P2Y1 receptor and a receptor previously classified as P2U. Here, we show that the P2U receptor is molecularly and pharmacologically identical to the cloned P2Y2 receptor. Both P2Y1 and P2Y2 receptors are necessary for full Ca2+ wave propagation in spinal astrocytes. Conversely, heterologous expression of either P2Y1 or P2Y2 receptors is sufficient for Ca2+ waves, and expressing these receptor subtypes together recapitulates the characteristics of Ca2+ waves in spinal astrocytes. Thus, P2Y1 and P2Y2 receptors are both necessary and sufficient for propagation of Ca2+ waves. Furthermore, we demonstrate that there are dramatic differences in the characteristics of Ca2+ waves propagating through each receptor subtype: Ca2+ waves propagating via P2Y2 receptors travel faster and further than those propagating via P2Y1 receptors. We find that the nucleotidase apyrase selectively blocks Ca2+ wave propagation through P2Y2 receptors but accelerates Ca2+ waves propagating through P2Y1 receptors. Taking our results together with those from the literature, we suggest that mediation of Ca2+ waves by ATP leading to activation of two subtypes of receptor, P2Y1 and P2Y2, may be a general principle for gliotransmission in the CNS. Thus, processes that alter expression or function of these receptors may control the rate and extent of astrocyte Ca2+ waves. PMID- 12890767 TI - Making memories without trying: medial temporal lobe activity associated with incidental memory formation during recognition. AB - Structures in the medial portions of the human temporal lobes (MTL) play a vital role in the ability to learn new facts and events, whether such learning is intentional or incidental. We examined neural activity in the MTL both while participants studied pictures of novel scenes and while they attempted to recognize which scenes had been previously presented. In a second surprise test we assessed participants' memory for items that were presented only during the previous recognition memory test. We present a novel approach to cross participant alignment of neuroimaging data that provides more precise localization and enhanced statistical power within regions such as the MTL. Using this technique, we observed that the amount of MTL activity predicted participants' ability to subsequently remember scenes not only during the intentional study task, but also during the first memory retrieval test when only incidental encoding occurred. This encoding-related activity during memory retrieval was in the same subregions of the MTL as encoding-related activity during intentional study and is hypothesized to be one of the primary reasons why retrieval-related activity is often difficult to observe with neuroimaging techniques. PMID- 12890766 TI - Beta-amyloid regulation of presynaptic nicotinic receptors in rat hippocampus and neocortex. AB - Alteration by beta-amyloid (Abeta) of signaling via nicotinic acetylcholine receptors (nAChRs) has been implicated in the early stages of Alzheimer's disease. nAChRs function both post- and presynaptically in the nervous system; however, little is known about the functional consequence of the interaction of Abeta with these receptors, particularly those on presynaptic nerve terminals. In view of the strong correlation between loss of synaptic terminals and dementia, together with the reduction in nAChRs in Alzheimer's disease, the possibility exists that presynaptic nAChRs may be targets for Abeta. To explore this possibility, we assessed the effect of Abeta peptides on nicotine-evoked changes in presynaptic Ca2+ level via confocal imaging of isolated presynaptic nerve endings from rat hippocampus and neocortex. Abeta1-42 appeared to inhibit presynaptic nAChR activation by nicotine. Surprisingly, picomolar Abeta1-42 was found to directly evoke sustained increases in presynaptic Ca2+ via nAChRs, revealing that the apparent inhibitory action of Abeta1-42 was the result of an occlusion of nicotine to further stimulate the receptors. The direct effect of Abeta was found to be sensitive to alpha-bungarotoxin, mecamylamine, and dihydro beta-erythroidine, indicating involvement of alpha7-containing nAChRs and non alpha7-containing nAChRs. Prior depolarization strongly attenuated subsequent Abeta-evoked responses in a manner dependent on the amplitude of the initial presynaptic Ca2+ increase, suggesting that nerve activity or Ca2+ channel density may control the impact of Abeta on presynaptic nerve terminal function. Together, these results suggest that the sustained increases in presynaptic Ca2+ evoked by Abeta may underlie disruptions in neuronal signaling via nAChRs in the early stages of Alzheimer's disease. PMID- 12890768 TI - Post-training intra-basolateral amygdala infusions of norepinephrine enhance consolidation of memory for contextual fear conditioning. AB - Post-training infusions of drugs, including noradrenergic agonists and antagonists, into the basolateral amygdala (BLA) influence the consolidation of memory for training in several tasks, including inhibitory avoidance. There is, however, conflicting evidence concerning whether post-training intra-BLA drug infusions modulate the consolidation of contextual fear conditioning (CFC). In the present study, norepinephrine (NE) was infused bilaterally into the BLA of male Sprague Dawley rats immediately after training on two CFC tasks: a Y-maze and a straight alley. Post-training intra-BLA infusions enhanced memory of CFC training in the Y-maze, as assessed by percentage of time spent freezing and shock arm entrance latencies. Post-training intra-BLA infusions of NE enhanced 48 hr retention of CFC training in the straight alley, as assessed by shock compartment entrance latencies and the number of shocks required to learn to avoid entering the shock compartment. These findings indicate that the consolidation of memory for CFC, like that for inhibitory avoidance training, is influenced by post-training neuromodulatory influences within the BLA. Thus, the findings provide additional evidence consistent with the hypothesis that the BLA has a general role in modulating memory consolidation. PMID- 12890769 TI - Reversible suppression of glutamatergic neurotransmission of cerebellar granule cells in vivo by genetically manipulated expression of tetanus neurotoxin light chain. AB - We developed a novel technique that allowed reversible suppression of glutamatergic neurotransmission in the cerebellar network. We generated two lines of transgenic mice termed Tet and TeNT mice and crossed the two transgenic lines to produce the Tet/TeNT double transgenic mice. In the Tet mice, the tetracycline controlled reverse activator (rtTA) was expressed selectively in cerebellar granule cells by the promoter function of the GABA(A) receptor alpha6 subunit gene. In the TeNT mice, the fusion gene of tetanus neurotoxin light chain (TeNT) and enhanced green fluorescent protein (EGFP) was designed to be induced by the interaction of doxycycline (DOX)-activated rtTA with the tetracycline-responsive promoter. The Tet/TeNT mice grew normally even after DOX treatment and exhibited a restricted DOX-dependent expression of TeNT in cerebellar granule cells. Along with this expression, TeNT proteolytically cleaved the synaptic vesicle protein VAMP2 (also termed synaptobrevin2) and reduced glutamate release from granule cells. Both cleavage of VAMP2/synaptobrevin2 and reduction of glutamate release were reversed by removal of DOX. Among the four genotypes generated by heterozygous crossing of Tet and TeNT mice, only Tet/TeNT mice showed DOX dependent reversible motor impairments as analyzed with fixed bar and rota-rod tests. Reversible suppression of glutamatergic neurotransmission thus can be manipulated with spatiotemporal accuracy by DOX treatment and removal. These transgenic mice will serve as an animal model to study the cerebellar function in motor coordination and learning. PMID- 12890770 TI - Gap junctional coupling underlies the short-latency spike synchrony of retinal alpha ganglion cells. AB - We examined whether coupling between neighboringalpha-type ganglion cells (alpha GCs) in the rabbit retina underlies their synchronous spike activity. Simultaneous recordings were made from arrays of alpha-GCs to determine the synchrony of both spontaneous and light-evoked spike activity. One cell within each array was then injected with the biotinylated tracer Neurobiotin to determine which of the cells were coupled via gap junctions. Cross-correlation analyses indicated that neighboring off-center alpha-GCs maintain short-latency (approximately 2.5 msec) synchronous spiking, whereas the spontaneous spike activities of on-centeralpha-GC neighbors are not correlated. Without exception, those off-centeralpha-GCs showing synchronous spiking were found to be tracer coupled to both amacrine cells and neighboring off-centeralpha-GCs. In contrast, on-center alpha-GCs were never tracer coupled. Furthermore, whereas spikes initiated in an off-center alpha-GC with extrinsic current injection resulted in short-latency synchronized spiking in neighboring off-center alpha-GCs, this was never seen between on-center alpha-GCs. These results indicate that electrical coupling via gap junctions underlies the short-latency concerted spike activity of neighboring alpha-GCs. PMID- 12890771 TI - The relay of high-frequency sensory signals in the Whisker-to-barreloid pathway. AB - The present study investigated the operational features of whisker-evoked EPSPs in barreloid cells and the ability of the whisker-to-barreloid pathway to relay high rates of whisker deflection in lightly anesthetized rats. Results show that lemniscal EPSPs are single-fiber events with fast rise times (<500 microsec) that strongly depress at short inter-EPSP intervals. They occur at short latencies (3.84 +/- 0.96 msec) with little jitters (<300 microsec) after electrical stimulation of the whisker follicle. Waveform analysis indicates that one to three lemniscal axons converge on individual barreloid cells to produce EPSPs of similar rise times but different amplitudes. When challenged by high rates of whisker deflection, cells in the whisker-to-barreloid pathway demonstrate a remarkable frequency-following ability. Primary vibrissa afferents could follow in a phase-locked manner trains of sinusoidal deflections at up to 1 kHz. Although trigeminothalamic cells could still faithfully follow deflection rates of 200-300 Hz, the actual frequency-following ability of individual cells depends on the amplitude, velocity, and direction of displacements. The discharges of trigeminothalamic cells induce corresponding phase-locked EPSPs in barreloid cells, which trigger burst discharges at stimulus onset. During the following cycles of the stimulus train, few action potentials ensue because of the strong synaptic depression at lemniscal synapses. It is concluded that the whisker-to barreloid pathway can relay vibratory inputs with a high degree of temporal precision, but that the relay of this information to the cerebral cortex requires the action of modulators, and possibly phase-locked discharges among an ensemble of relay cells. PMID- 12890772 TI - App gene dosage modulates endosomal abnormalities of Alzheimer's disease in a segmental trisomy 16 mouse model of down syndrome. AB - Altered neuronal endocytosis is the earliest known pathology in sporadic Alzheimer's disease (AD) and Down syndrome (DS) brain and has been linked to increased Abeta production. Here, we show that a genetic model of DS (trisomy 21), the segmental trisomy 16 mouse Ts65Dn, develops enlarged neuronal early endosomes, increased immunoreactivity for markers of endosome fusion (rab5, early endosomal antigen 1, and rabaptin5), and endosome recycling (rab4) similar to those in AD and DS individuals. These abnormalities are most prominent in neurons of the basal forebrain, which later develop aging-related atrophy and degenerative changes, as in AD and DS. We also show that App, one of the triplicated genes in Ts65Dn mice and human DS, is critical to the development of these endocytic abnormalities. Selectively deleting one copy of App or a small portion of the chromosome 16 segment containing App from Ts65Dn mice eliminated the endosomal phenotype. Overexpressing App at high levels in mice did not alter early endosomes, implying that one or more additional genes on the triplicated segment of chromosome 16 are also required for the Ts65Dn endosomal phenotype. These results identify an essential role for App gene triplication in causing AD related endosomal abnormalities and further establish the pathogenic significance of endosomal dysfunction in AD. PMID- 12890773 TI - N-type calcium channel alpha1B subunit (Cav2.2) knock-out mice display hyperactivity and vigilance state differences. AB - Differential properties of voltage-dependent Ca2+ channels have been primarily ascribed to the alpha1 subunit, of which 10 different subtypes are currently known. For example, channels that conduct the N-type Ca2+ current possess the alpha1B subunit (Cav2.2), which has been localized, inter alia, to the piriform cortex, hippocampus, hypothalamus, locus coeruleus, dorsal raphe, thalamic nuclei, and granular layer of the cortex. Some of these regions have been previously implicated in metabolic and vigilance state control, and selective block of the N-type Ca2+ channel causes circadian rhythm disruption. In this study of Cav2.2-/- knock-out mice, we examined potential differences in feeding behavior, spontaneous locomotion, and the sleep-wake cycle. Cav2.2-/- mice did not display an overt metabolic phenotype but were hyperactive, demonstrating a 20% increase in activity under novel conditions and a 95% increase in activity under habituated conditions during the dark phase, compared with wild-type littermates. Cav2.2-/- mice also displayed vigilance state differences during the light phase, including increased consolidation of rapid-eye movement (REM) sleep and increased intervals between non-REM (NREM) and wakefulness episodes. EEG spectral power was increased during wakefulness and REM sleep and was decreased during NREM sleep in Cav2.2-/- mice. These results indicate a role of the N-type Ca2+ channel in activity and vigilance state control, which we interpret in terms of effects on neurotransmitter release. PMID- 12890774 TI - Coherent oscillations in neuronal activity of the supplementary motor area during a visuomotor task. AB - Neural activity recorded in behaving animals is nonstationary, making it difficult to determine factors influencing its temporal patterns. In the present study, rhesus monkeys were trained to produce a series of visually guided hand movements according to the changes in target locations, and multichannel single neuron activity was recorded from the caudal supplementary motor area. Coherent oscillations in neural activity were analyzed using the wavelet cross-spectrum, and its statistical significance was evaluated using various methods based on surrogate spike trains and trial shuffling. A population-averaged wavelet cross spectrum displayed a strong tendency for oscillatory activity in the gamma frequency range (30 approximately 50 Hz) to synchronize immediately before and after the onset of movement target. The duration of synchronized oscillations in the gamma frequency range increased when the onset of the next target was delayed. In addition, analysis of individual neuron pairs revealed that many neuron pairs also displayed coherent oscillations in the beta frequency range (15 30 Hz). Coherent beta frequency oscillations were less likely to be synchronized than gamma frequency oscillations, consistent with the fact that coherent beta frequency oscillations were not clearly seen in the population-averaged cross spectrum. For a given neuron pair, the time course and phase of coherent oscillations were often similar across different movements. These results are consistent with the proposal that synchronized oscillations in the gamma frequency range might be related to the anticipation of behaviorally relevant events and the contextual control of cortical information flow. PMID- 12890775 TI - The human immunodeficiency virus-1 protein Tat and its discrete fragments evoke selective release of acetylcholine from human and rat cerebrocortical terminals through species-specific mechanisms. AB - The effect of the human immunodeficiency virus-1 protein Tat was investigated on neurotransmitter release from human and rat cortical nerve endings. Tat failed to affect the release of several neurotransmitters, such as glutamate, GABA, norepinephrine, and others, but it evoked the release of [3H]ACh via increase of cytosolic [Ca2+]. In human nerve terminals, the Tat effect partly depends on Ca2+ entry through voltage-sensitive Ca2+ channels, because Cd2+ halved the Tat-evoked release. Activation of group I metabotropic glutamate receptors (mGluR) and mobilization of Ca2+ from IP3-sensitive intraterminal stores are also involved, because the Tat effect was prevented by mGluR antagonists 2-methyl-6 (phenylethynyl)pyridine hydrochloride and 7-(hydroxyimino)cyclopropa[b]chromen-1a carboxylate ethyl ester and by the IP3 receptor antagonists heparin and xestospongin C. Furthermore, the group I selective mGlu agonist (RS)-3,5 dihydroxyphenylglycine enhanced [3H]ACh release. In rat nerve terminals, the Tat evoked release neither depends on external Ca2+ ions entry nor on IP3-mediated mechanisms. Tat seems to cause mobilization of Ca2+ from ryanodine-sensitive internal stores because its effect was prevented by both 8-bromo-cyclic adenosine diphosphate-ribose and dantrolene. The Tat-evoked release from human synaptosomes was mimicked by the peptide sequences Tat 32-62, Tat 49-86, and Tat 41-60. In contrast, the Tat 49-86 and Tat 61-80 fragments, but not the Tat 32-62 fragment, were active in rat synaptosomes. In conclusion, Tat elicits Ca2+-dependent [3H]ACh release by species-specific intraterminal mechanisms by binding via discrete amino acid sequences to different receptive sites on human and rat cholinergic terminals. PMID- 12890776 TI - Brain activity evoked by the perception of human walking: controlling for meaningful coherent motion. AB - Many functional neuroimaging studies of biological motion have used as stimuli point-light displays of walking figures and compared the resulting activations with those evoked by the same display elements moving in a random or noncoherent manner. Although these studies have established that biological motion activates the superior temporal sulcus (STS), the use of random motion controls has left open the possibility that coordinated and meaningful nonbiological motion might activate these same brain regions and thus call into question their specificity for processing biological motion. Here we used functional magnetic resonance imaging and an anatomical region-of-interest approach to test a hierarchy of three questions regarding activity within the STS. First, by comparing responses in the STS with animations of human and robot walking figures, we determined (1) that the STS is sensitive to biological motion itself, not merely to the superficial characteristics of the stimulus. Then we determined that the STS responds more strongly to biological motion (as conveyed by the walking robot) than to (2) a nonmeaningful but complex nonbiological motion (a disjointed mechanical figure) and (3) a complex and meaningful nonbiological motion (the movements of a grandfather clock). In subsequent whole-brain voxel-based analyses, we confirmed robust STS activity that was strongly right lateralized. In addition, we observed significant deactivations in the STS that differentiated biological and nonbiological motion. These voxel-based analyses also revealed regions of motion-related positive activity in other brain regions, including MT or V5, fusiform gyri, right premotor cortex, and the intraparietal sulci. PMID- 12890778 TI - Neurokinin-1 projection cells in the rat dorsal horn receive synaptic contacts from axons that possess alpha2C-adrenergic receptors. AB - Thealpha2C subclass of adrenergic receptor (alpha2C-AR) mediates some of the antinociceptive actions of norepinephrine in the spinal cord. Axon terminals, which possess this receptor, are concentrated in the superficial dorsal horn and originate from spinal interneurons. We performed a series of combined tract tracing and immunocytochemical studies to determine whether alpha2C-AR immunoreactive axons target projection neurons that possess the neurokinin-1 (NK 1) receptor because such cells are likely to transmit nociceptive information to the brain. Spinomedullary neurons were labeled by stereotaxic injection of the B subunit of cholera toxin (CTb) into the caudal ventrolateral medulla of three anesthetized adult rats. After 3 d, the animals were anesthetized again and fixed by perfusion. Sections were cut from midlumbar segments and reacted with antibodies to reveal alpha2C-ARs, CTb, and NK-1 receptors. Retrogradely labeled neurons possessing the NK-1 receptor (n = 45) were examined with confocal microscopy to investigate their relationship with alpha2C-AR-immunoreactive axons. Numerous alpha2C-AR axons were apposed to cell bodies and proximal dendrites of cells in lamina I and also to distal dendrites that originate from labeled cell bodies in lamina III/IV. A combined confocal and electron microscopic method confirmed that these appositions were synaptic. Additional experiments showed that virtually all alpha2C-AR terminals in contact with labeled cells are also immunoreactive for the vesicular glutamate transporter 2 and therefore are glutamatergic. These data suggest that norepinephrine can modulate excitatory synaptic transmission from spinal interneurons to projection cells by acting at alpha2C-ARs. This could be one of the mechanisms that underlie the antinociceptive actions of norepinephrine. PMID- 12890777 TI - Enhanced expression of a specific hyperpolarization-activated cyclic nucleotide gated cation channel (HCN) in surviving dentate gyrus granule cells of human and experimental epileptic hippocampus. AB - Changes in the expression of ion channels, contributing to altered neuronal excitability, are emerging as possible mechanisms in the development of certain human epilepsies. In previous immature rodent studies of experimental prolonged febrile seizures, isoform-specific changes in the expression of hyperpolarization activated cyclic nucleotide-gated cation channels (HCNs) correlated with long lasting hippocampal hyperexcitability and enhanced seizure susceptibility. Prolonged early-life seizures commonly precede human temporal lobe epilepsy (TLE), suggesting that transcriptional dysregulation of HCNs might contribute to the epileptogenic process. Therefore, we determined whether HCN isoform expression was modified in hippocampi of individuals with TLE. HCN1 and HCN2 expression were measured using in situ hybridization and immunocytochemistry in hippocampi from three groups: TLE with hippocampal sclerosis (HS; n = 17), epileptic hippocampi without HS, or non-HS (NHS; n = 10), and autopsy material (n = 10). The results obtained in chronic human epilepsy were validated by examining hippocampi from the pilocarpine model of chronic TLE. In autopsy and most NHS hippocampi, HCN1 mRNA expression was substantial in pyramidal cell layers and lower in dentate gyrus granule cells (GCs). In contrast, HCN1 mRNA expression over the GC layer and in individual GCs from epileptic hippocampus was markedly increased once GC neuronal density was reduced by >50%. HCN1 mRNA changes were accompanied by enhanced immunoreactivity in the GC dendritic fields and more modest changes in HCN2 mRNA expression. Furthermore, similar robust and isoform selective augmentation of HCN1 mRNA expression was evident also in the pilocarpine animal model of TLE. These findings indicate that the expression of HCN isoforms is dynamically regulated in human as well as in experimental hippocampal epilepsy. After experimental febrile seizures (i.e., early in the epileptogenic process), the preserved and augmented inhibition onto principal cells may lead to reduced HCN1 expression. In contrast, in chronic epileptic HS hippocampus studied here, the profound loss of interneuronal and principal cell populations and consequent reduced inhibition, coupled with increased dendritic excitation of surviving GCs, might provoke a "compensatory" enhancement of HCN1 mRNA and protein expression. PMID- 12890779 TI - Evidence for chelatable zinc in the extracellular space of the hippocampus, but little evidence for synaptic release of Zn. AB - Zinc colocalizes with glutamate in the synaptic vesicles of certain glutamatergic vesicles in the mammalian brain. Here, I introduce a method for detecting Zn in the extracellular space of brain slices and another method for detecting the passage of Zn out of the slice. In both cases, the fluorimetric Zn probe FluoZin 3 is used in conjunction with a slow Zn chelator, Ca-EDTA, to reduce background fluorescence. In addition, a new Zn chelator, ethylenediiminodi-2-pentanedioic acid, with little affinity for Ca or Mg is introduced. These tools are then used to show that little Zn (approximately 2 nm) is released during the course of synaptic transmission into the extracellular space. However, when hippocampal slices are subjected to a high potassium stimulus (50 mM) combined with an increase in osmolarity, Zn is externalized in the Timm's-stained areas (approximately 6 nm). This stimulus also leads to even greater Zn elevations in area CA1 that is only weakly stained by the Timm's method. Nevertheless, even under these conditions, little if any Zn makes its way out of the slices. I present evidence for a layer of Zn in the extracellular space that maps onto the Timm's stained region of the hippocampus. This Zn veneer appears to be loosely associated with molecules in the extracellular space and may be the raison d'etre for vesicular Zn. PMID- 12890780 TI - Brain-derived neurotrophic factor is required for the maintenance of cortical dendrites. AB - Brain-derived neurotrophic factor (BDNF) is thought to be involved in neuronal survival, migration, morphological and biochemical differentiation, and modulation of synaptic function in the CNS. In the rodent cortex, postnatal BDNF expression is initially low but subsequently increases to reach maximal levels around weaning. Thus, BDNF expression peaks at a time when both structural and functional maturation of cortical circuitry occurs. Although the function of BDNF has been probed using many approaches, its requirements during this phase of life have not previously been examined genetically. To test the in vivo requirements for BDNF during this important phase of development we generated early-onset forebrain-specific BDNF mutant mice. Although these mice undergo forebrain restricted deletion of BDNF by Cre-mediated recombination during embryogenesis, they are healthy, and we did not detect the loss of specific cortical excitatory or inhibitory neurons. However, the neocortex of 5-week-old mice was thinner, attributable at least partly to neuronal shrinkage. Importantly, although visual cortical layer 2/3 neurons in the mutants initially developed normal dendrite structure, dendritic retraction became apparent by 3 weeks of age. Thus, our observations suggest that cortically expressed BDNF functions to support the maintenance of cortical neuron size and dendrite structure rather than the initial development of these features. This is consistent with a role for BDNF in stabilizing the "survival" of circuitry during the phase of activity-dependent reorganization of cortical connectivity. PMID- 12890781 TI - Axonal dopamine receptors activate peripheral spike initiation in a stomatogastric motor neuron. AB - We studied the effects of dopamine on the stomatogastric ganglion (STG) of the lobster, Homarus americanus. The two pyloric dilator (PD) neurons are active in the pyloric rhythm, have somata in the STG, and send axons many centimeters to innervate muscles of the stomach. Dopamine application to the stomatogastric nervous system when the PD neurons were rhythmically active evoked additional action potentials during the PD neuron interburst intervals. These action potentials were peripherally generated at a region between the STG and the first bilateral branch, approximately 1 cm away from the STG, and traveled antidromically to the neuropil and orthodromically to the pyloric dilator muscles. Focal applications of dopamine to the nerves showed that spikes could be initiated in almost the entire peripheral axon of the PD neurons. Dopamine also evoked spikes in isolated peripheral axons. The concentration threshold for peripheral spike initiation was at or below 10-9 m dopamine. Thus, the peripheral axon can play an important role in shaping the output signaling to the muscles by the motor neuron. PMID- 12890782 TI - Development and subunit composition of synaptic NMDA receptors in the amygdala: NR2B synapses in the adult central amygdala. AB - NMDA receptors are well known to play an important role in synaptic development and plasticity. Functional NMDA receptors are heteromultimers thought to contain two NR1 subunits and two or three NR2 subunits. In central neurons, NMDA receptors at immature glutamatergic synapses contain NR2B subunits and are largely replaced by NR2A subunits with development. At mature synapses, NMDA receptors are thought to be multimers that contain either NR1/NR2A or NR1/NR2A/NR2B subunits, whereas receptors that contain only NR1/NR2B subunits are extrasynaptic. Here, we have studied the properties of NMDA receptors at glutamatergic synapses in the lateral and central amygdala. We find that NMDA receptor-mediated synaptic currents in the central amygdala in both immature and mature synapses have slow kinetics and are substantially blocked by the NR2B selective antagonists (1S, 2S)-1-(4-hydroxyphenyl)-2-(4-hydroxy-4 phenylpiperidino)-1-propano and ifenprodil, indicating that there is no developmental change in subunit composition. In contrast, at synapses on pyramidal neurons in the lateral amygdala, whereas NMDA EPSCs at immature synapses are slow and blocked by NR2B-selective antagonists, at mature synapses their kinetics are faster and markedly less sensitive to NR2B-selective antagonists, consistent with a change from NR2B to NR2A subunits. Using real-time PCR and Western blotting, we show that in adults the ratio of levels of NR2B to NR2A subunits is greater in the central amygdala than in the lateral amygdala. These results show that the subunit composition synaptic NMDA receptors in the lateral and central amygdala undergo distinct developmental changes. PMID- 12890784 TI - Presynaptic calcium stores modulate afferent release in vestibular hair cells. AB - Hair cells, the mechanoreceptors of the acoustic and vestibular system, are presynaptic to primary afferent neurons of the eighth nerve and excite neural activity by the release of glutamate. In the present work, the role played by intracellular Ca2+ stores in afferent transmission was investigated, at the presynaptic level, by monitoring changes in the intracellular Ca2+ concentration ([Ca2+]i) in vestibular hair cells, and, at the postsynaptic level, by recording from single posterior canal afferent fibers. Application of 1-10 mm caffeine to hair cells potentiated Ca2+ responses evoked by depolarization at selected Ca2+ hot spots, and also induced a graded increase in cell membrane capacitance (DeltaCm), signaling exocytosis of the transmitter. Ca2+ signals evoked by caffeine peaked in a region located approximately 10 microm from the base of the hair cell. [Ca2+]i increases, similarly localized, were observed after 500 msec depolarizations, but not with 50 msec depolarizations, suggesting the occurrence of calcium-induced calcium release (CICR) from the same stores. Both Ca2+ and DeltaCm responses were inhibited after incubation with ryanodine (40 microm) for 8-10 min. Consistent with these results, afferent transmission was potentiated by caffeine and inhibited by ryanodine both at the level of action potentials and of miniature EPSPs (mEPSPs). Neither caffeine nor ryanodine affected the shape and amplitude of mEPSPs, indicating that both drugs acted at the presynaptic level. These results strongly suggest that endogenous modulators of the CICR process will affect afferent activity elicited by mechanical stimuli in the physiological frequency range. PMID- 12890783 TI - Response suppression in v1 agrees with psychophysics of surround masking. AB - When a target stimulus is embedded in a high contrast surround, the target appears reduced in contrast and is harder to detect, and neural responses in visual cortex are suppressed. We used functional magnetic resonance imaging (fMRI) and psychophysics to quantitatively compare these physiological and perceptual effects. Observers performed a contrast discrimination task on a contrast-reversing sinusoidal target grating. The target was either presented in isolation or embedded in a high-contrast surround. While observers performed the task, we also measured fMRI responses as a function of target contrast, both with and without a surround. We found that the surround substantially increased the psychophysical thresholds while reducing fMRI responses. The two data sets were compared, on the basis of the assumption that a fixed response difference is required for correct discrimination, and we found that the psychophysics accounted for 96.5% of the variance in the measured V1 responses. The suppression in visual areas V2 and V3 was stronger, too strong to agree with psychophysics. The good quantitative agreement between psychophysical thresholds and V1 responses suggests V1 as a plausible candidate for mediating surround masking. PMID- 12890785 TI - AII amacrine cells express L-type calcium channels at their output synapses. AB - AII amacrine cells play a critical role in the high-fidelity signal transmission pathways involved with nighttime vision. The temporal properties of the light responses strongly depend on the transfer function at different synaptic stages and consequently on presynaptic calcium influx. AII light responses are complex waveforms generated by graded input, they comprise Na+-based spikes as well as a sustained component, and they are transferred to graded cone bipolar cells. It is, therefore, of interest to determine the properties of AII voltage-dependent calcium channels (VDCCs) to establish whether these cells express N-type and/or P/Q-type VDCCs, characteristic of spiking neurons, or whether they are more like graded neurons, which mostly use L-type VDCCs. We combined electrophysiological, molecular biological, and imaging techniques to characterize calcium currents and their sites of origin in mouse AII amacrine cells. Calcium currents activated at potentials more positive than -60 mV (maximally between -50 and -20 mV) and inactivated slowly. These currents were blocked by dihydropyridine (DHP) antagonists and were enhanced by the DHP agonist BayK 8644. Single-cell RT-PCR analysis of mRNA encoding for different calcium channel alpha subunits in AIIs revealed a consistent expression of the alpha1-D subunit. Calcium imaging of AII cells showed that the greatest change in intracellular calcium occurred in the lobular appendages, with minor changes being observed in the arboreal dendrites. Depolarization-induced calcium rises were also modulated by DHPs, suggesting that a particular kind of L-type VDCC, mainly localized to the lobular appendages, enables these spiking-capable neurons to release neurotransmitter in a sustained manner onto OFF-cone bipolar cells. PMID- 12890786 TI - DNA synthesis and neuronal apoptosis caused by familial Alzheimer disease mutants of the amyloid precursor protein are mediated by the p21 activated kinase PAK3. AB - Apoptotic pathways and DNA synthesis are activated in neurons in the brains of individuals with Alzheimer disease (AD). However, the signaling mechanisms that mediate these events have not been defined. We show that expression of familial AD (FAD) mutants of the amyloid precursor protein (APP) in primary neurons in culture causes apoptosis and DNA synthesis. Both the apoptosis and the DNA synthesis are mediated by the p21 activated kinase PAK3, a serine-threonine kinase that interacts with APP. A dominant-negative kinase mutant of PAK3 inhibits the neuronal apoptosis and DNA synthesis; this effect is abolished by deletion of the PAK3 APP-binding domain or by coexpression of a peptide representing this binding domain. The involvement of PAK3 specifically in FAD APP mediated apoptosis rather than in general apoptotic pathways is suggested by the facts that a dominant-positive mutant of PAK3 does not alone cause neuronal apoptosis and that the dominant-negative mutant of PAK3 does not inhibit chemically induced apoptosis. Pertussis toxin, which inactivates the heterotrimeric G-proteins Go and Gi, inhibits the apoptosis and DNA synthesis caused by FAD APP mutants; the apoptosis and DNA synthesis are rescued by coexpression of a pertussis toxin-insensitive Go. FAD APP-mediated DNA synthesis precedes FAD APP-mediated apoptosis in neurons, and inhibition of neuronal entry into the cell cycle inhibits the apoptosis. These data suggest that a normal signaling pathway mediated by the interaction of APP, PAK3, and Go is constitutively activated in neurons by FAD mutations in APP and that this activation causes cell cycle entry and consequent apoptosis. PMID- 12890787 TI - Long memory in song learning by zebra finches. AB - Young songbirds use memorized tutor songs as templates to shape their own songs. This process requires control of voice by auditory feedback. We prevented zebra finches from hearing their own vocalizations by exposure to loud noise after 35 d of age, before which they had been reared with song tutors from birth. When the noise stopped at 102-200 d of age, the birds sang unstable and noisy song syllables that did not resemble the tutor syllables. The similarity to the tutor syllables steadily increased until the time of song crystallization approximately 30 d later. These findings show that the memory of tutor syllables survives auditory perturbations during the period when it is normally recalled and that zebra finches can use the memory well after the normal period of song development. The temporal order of syllables resembled the tutor model only in birds released from the noise before 80 d of age but not in older birds. Thus, different schedules and processes may govern the learning of syllable phonology and syntax. PMID- 12890788 TI - Response to contrast of electrophysiologically defined cell classes in primary visual cortex. AB - Information processing in the visual cortex is critically dependent on the input output relationships of its component neurons. The transformation of synaptic inputs into spike trains depends in turn on the host of intrinsic membrane properties expressed by neurons, which define established electrophysiological cell classes in the neocortex. Here we studied, with intracellular recordings in vivo, how the electrophysiological cell classes in the primary visual cortex transform an increasing input, represented by stimulus contrast, into membrane depolarization and trains of action potentials. We used contrast as input because, regardless of their stimulus selectivity, primary visual cortical cells increase their firing rates in response to increases in luminance contrast. We found that both the spike rate response and the membrane potential response are best described by the hyperbolic ratio function when compared with linear, power, and logarithmic functions. In addition, both responses show similar parameter values and similar residual variance from the fits to all four functions. We also found that changes in membrane potential are similar, but firing rates differ strongly, between the established electrophysiological cell classes: fast spiking neurons show the highest firing rates, followed by fast rhythmic bursting, and regular spiking (RS) cells. In addition, among complex cells, RS cells from supragranular layers fired at higher rates than RS cells from infragranular layers. Finally, we show that the differences in firing rates between cell classes arise from differences in the slope of the relationship between membrane potential and spike rate. PMID- 12890789 TI - Olfactory bulb mitral-tufted cell plasticity: odorant-specific tuning reflects previous odorant exposure. AB - Olfactory system second-order neurons, mitral-tufted cells, have odorant receptive fields (ORFs) (molecular receptive ranges in odorant space for carbon chain length in organic odorant molecules). This study quantified several dimensions of these excitatory odorant receptive fields to novel odorants in rats and then examined the effects of passive odorant exposure on the shape of the ORF tuning curve. ORFs for carbon chain length of novel ethyl esters (pure odorants that the animals had not been exposed to previously) were determined before and after a 50 sec prolonged exposure to one of the odorants. In response to novel odorants, quantitative analysis of mitral-tufted cell excitatory ORFs revealed that the median ORF width spanned 3-4 carbons, generally with a single-most excitatory odorant. Exposure to either the most excitatory odorant (ON-PEAK) or an odorant that was two carbons longer (OFF-PEAK) for 50 sec produced whole ORF suppression immediately after the end of the prolonged exposure, with the ON-PEAK exposure producing the greatest suppression. These results are consistent with a feature-detecting function for mitral-tufted cells. Redetermination of the ORF 15 and 60 min after the exposure revealed that OFF-PEAK exposure produced a reduction in responsiveness to the best odorant and an increase in responsiveness to the exposed odorant. In contrast, exposure to the ON-PEAK odorant or no odorant did not affect ORFs. Given that mitral-tufted cells receive exclusive excitatory input from olfactory receptor neurons expressing identical receptor proteins, it is hypothesized that experience-induced mitral-tufted cell ORF changes reflect modulation of lateral and centrifugal olfactory bulb circuits. PMID- 12890790 TI - Lack of huntingtin-associated protein-1 causes neuronal death resembling hypothalamic degeneration in Huntington's disease. AB - Huntington's disease (HD) is caused by a polyglutamine expansion in the disease protein huntingtin. The polyglutamine expansion causes huntingtin to interact abnormally with a number of proteins. However, it is unclear whether, and how, huntingtin-associated proteins are involved in the neurodegeneration in HD. Here, we show that huntingtin-associated protein-1 (HAP1), which is involved in intracellular trafficking of epidermal growth factor receptor (EGFR), is highly expressed in the hypothalamus. Mice lacking HAP1 die after birth because of depressed feeding activity. Terminal deoxynucleotidyl transferase-mediated biotinylated UTP nick end labeling staining and electron microscopic examination revealed the degeneration in hypothalamic regions that control feeding behavior. Hypothalamic degeneration was also observed in HD transgenic mice that have a significant loss of body weight. Inhibition of HAP1 expression decreases EGFR signaling and cell viability, whereas overexpression of HAP1 enhances this signaling activity and inhibits mutant huntingtin-mediated cytotoxicity. These results suggest that the effect of mutant huntingtin on HAP1 and EGFR signaling may contribute to the hypothalamic neurodegeneration and loss of body weight in HD. PMID- 12890791 TI - Promiscuous and reversible blocker of presynaptic calcium channels in frog and crayfish neuromuscular junctions from Phoneutria nigriventer spider venom. AB - Peptide channel blockers found in venoms of many predators are useful pharmacological tools and potential therapeutic agents. The venom of the Brazilian spider Phoneutria nigriventer contains a fraction, omega-phonetoxin-IIA (omega-Ptx-IIA, 8360 MW), which blocks Ca2+ channels. At frog neuromuscular junctions (NMJ) bathed in normal Ca2+ (1.8 mM) saline, omega-Ptx IIA did not affect spontaneous transmitter release but reversibly reduced evoked transmitter release by 75 and 95% at 12 and 24 nM, respectively. In contrast, toxin effects were irreversible in low-Ca2+ (0.5 mM) saline. Ca2+ imaging in normal-Ca2+ saline showed that omega-Ptx-IIA partially blocked stimulus-dependent presynaptic Ca2+ signals, and the blockade was almost completely reversible. Increases in spontaneous release frequency induced by high extracellular K+ were blocked by omega-Ptx-IIA. Therefore omega-Ptx-IIA blocks N-type Ca2+ channels, which admit Ca2+ that triggers transmitter release at the frog NMJ. Additional evidence predicts that omega-Ptx-IIA binds to N-type Ca2+ channels at a different site from that of omega-Conotoxin-GVIA. omega-Ptx-IIA also gave a low-affinity partial blockade of transmitter release and presynaptic Ca2+ signals at crayfish NMJs where P-type channels are blocked by omega-agatoxin-IVA. The Ca2+-dependent reversibility and promiscuity of this toxin may make it highly useful experimentally and therapeutically. PMID- 12890792 TI - Effects of amplitude modulation on the coding of interaural time differences of low-frequency sounds in the inferior colliculus. I. Response properties. AB - Most sounds in the natural environment are amplitude-modulated (AM). To determine if AM alters the neuronal sensitivity to interaural time differences (ITDs) in low-frequency sounds, we tested neuronal responses to a binaural beat stimulus with and without modulation. We recorded from single units in the inferior colliculus of the unanesthetized rabbit. We primarily used low frequency ( approximately 25 Hz) modulation that was identical at both ears. We found that modulation could enhance, suppress, or not affect the discharge rate. In extreme cases, a neuron that showed no response to the unmodulated binaural beat did so when modulation was added to both ears. At the other extreme, a neuron that showed sensitivity to the unmodulated binaural beat ceased firing with modulation. Modulation could also affect the frequency range of ITD sensitivity, best ITD, and ITD tuning width. Despite these changes in individual neurons, averaging across all neurons, the peak and width of the population ITD function remained unchanged. Because ITD-sensitive neurons also time-locked to the modulation frequency, the location and sound attributes are processed simultaneously by these neurons. PMID- 12890793 TI - Effects of amplitude modulation on the coding of interaural time differences of low-frequency sounds in the inferior colliculus. II. Neural mechanisms. AB - In our companion paper, we reported on interaural time difference (ITD)-sensitive neurons that enhanced, suppressed, or did not change their response when identical AM was added to both ears. Here, we first examined physical factors such as the difference in the interaural correlation, spectrum, or energy between the modulated and unmodulated signals. These were insufficient to explain the observed enhancement and suppression. We then examined neural mechanisms by selectively modulating the signal to each ear, varying modulation depth, and adding background noise to the unmodulated signal. These experiments implicated excitatory and inhibitory monaural inputs to the inferior colliculus (IC). These monaural inputs are postulated to adapt to an unmodulated signal and adapt less to a modulated signal. Thus enhancement or suppression is created by the convergence of these excitatory or inhibitory inputs with the inputs from the binaural comparators. Under modulation, the role of the monaural input is to shift the threshold of the IC neuron. Consistent with this role, background noise mimicked the effect of modulation. Functionally, enhancement and suppression may serve in detecting the degree of modulation in a sound source while preserving ITD information. PMID- 12890794 TI - Serotonin reduces the hyperpolarization-activated current (Ih) in ventral tegmental area dopamine neurons: involvement of 5-HT2 receptors and protein kinase C. AB - Dopaminergic neurons of the ventral tegmental area (VTA) have been implicated in the rewarding properties of drugs of abuse and in the etiology of schizophrenia; serotonin modulation of these neurons may play a role in these phenomena. Whole cell patch-in-the-slice recording in rat brain slices was used to investigate modulation of the hyperpolarization-activated cationic current Ih by serotonin in these neurons. Serotonin (50-500 microM) reduced the amplitude of Ih in a concentration-dependent manner; this effect was reversible after prolonged washout of serotonin. This effect was mimicked by the 5-HT2 agonist alpha methylserotonin (25 microM) and reversed by the 5-HT2 antagonist ketanserin (25 microM). Serotonin reduced the maximal Ih current and conductance (measured at 130 mV) and caused a negative shift in the voltage dependence of Ih activation. The serotonin-induced reduction in Ih amplitude was antagonized by intracellular administration of the nonspecific protein kinase inhibitor H-7 (75 microM) and the selective protein kinase C inhibitor chelerythrine (25 microM). The protein kinase C activator phorbol 12, 13 diacetate (PDA, 2 microM) reduced Ih amplitude; when PDA and serotonin were applied together, the effect on Ih was less than additive. These data support the conclusion that serotonin reduces Ih in dopaminergic VTA neurons by acting at serotonin 5-HT2 receptors, which activate protein kinase C. This reduction of Ih may be physiologically important, as the selective inhibitor of Ih, ZD7288, significantly increased dopamine inhibition of firing rate of dopaminergic VTA neurons, an effect that we previously demonstrated with serotonin. PMID- 12890795 TI - Passive transport disrupts directional path integration by rat head direction cells. AB - A subset of neurons in the rat limbic system encodes head direction (HD) by selectively discharging when the rat points its head in a preferred direction in the horizontal plane. The preferred firing direction is sensitive to the location of landmark cues, as well as idiothetic or self-motion cues (i.e., vestibular, motor efference copy, proprioception, and optic flow). Previous studies have shown that the preferred firing direction remains relatively stable (average shift +/- 18 degrees ) after the rat walks from a familiar environment into a novel one, suggesting that without familiar landmarks, the preferred firing direction can be maintained using idiothetic cues, a process called directional path integration. This study repeated this experiment and manipulated the idiothetic cues available to the rat as it moved between the familiar and novel environment. Motor efference copy/proprioceptive cues were disrupted by passively transporting the animal between the familiar and novel environment. Darkening the room as the animal moved to the novel environment eliminated optic flow cues. HD cell preferred firing directions shifted in the novel environment by an average of 30 degrees after locomotion from the familiar environment with the room lights off; by an average of 70 degrees after passive transport from the familiar environment with the room lights on; and by an average of 67 degrees after passive transport with the room lights off. These findings are consistent with the view that motor efference copy/proprioception cues are important for maintaining the preferred firing direction of HD cells under conditions requiring path integration. PMID- 12890796 TI - Two distinct types of corticothalamic EPSPs and their contribution to short-term synaptic plasticity. AB - The lateral posterior nucleus (LPN) is innervated by two different morphological types of cortical terminals that originate from cortical layers V and VI. Here we describe two distinct types of excitatory postsynaptic potentials (EPSPs) that were recorded in the LPN after stimulation of corticothalamic fibers. These types of EPSPs differed in amplitude, latency, rise time, and response to increasing levels of stimulus intensity. The most frequently encountered EPSP, type I, displayed a longer latency and slower rise time than the less frequently encountered type II EPSP. Type I EPSPs also showed a graded increase in amplitude with increasing levels of stimulation, whereas type II EPSPs showed an all-or none response. In response to repetitive stimulation (0.5-20 Hz), type I EPSPs displayed frequency-dependent facilitation, whereas type II EPSPs displayed frequency-dependent depression. Further details of these distinct forms of short term synaptic plasticity were explored using paired-pulse stimuli. Pharmacology experiments revealed that both N-methyl-d-aspartate (NMDA) and non-NMDA glutamate receptors are involved in corticothalamic synaptic transmission in the LPN and contribute to both synaptic facilitation and depression. Taken together with the results of our previous anatomical studies, these results suggest that type I EPSPs arise from stimulation of layer VI afferents, whereas type II EPSPs arise from stimulation of layer V inputs. Moreover, type I and II EPSPs in the LPN may be functionally similar to corticogeniculate and retinogeniculate EPSPs, respectively. PMID- 12890797 TI - GABA(A) receptor activation modulates corneal unit activity in rostral and caudal portions of trigeminal subnucleus caudalis. AB - Corneal nociceptors terminate at the trigeminal subnucleus interpolaris/caudalis (Vi/Vc) transition and subnucleus caudalis/upper cervical spinal cord (Vc/C1) junction regions of the lower brain stem. The aims of this study were to determine if local GABAA receptor activation modifies corneal input to second order neurons at these regions and if GABAA receptor activation in one region affects corneal input to the other region. In barbiturate-anesthetized male rats, corneal nociceptors were excited by pulses of CO2 gas, and GABAA receptors were activated by microinjections of the selective agonist muscimol. Local muscimol injection at the site of recording inhibited all Vi/Vc and Vc/C1 units tested and was reversed partially by bicuculline. To test for ascending intersubnuclear communication, muscimol injection into the caudal Vc/C1 junction, remote from the recording site at the Vi/Vc transition, inhibited the evoked response of most corneal units, although some neurons were enhanced. Injection of the nonselective synaptic blocking agent, CoCl2, remotely into the Vc/C1 region inhibited the evoked response of all Vi/Vc units tested. To test for descending intersubnuclear communication, muscimol was injected remotely into the rostral Vi/Vc transition and enhanced the evoked activity of all corneal units tested at the caudal Vc/C1 junction. These results suggest that GABAA receptor mechanisms play a significant role in corneal nociceptive processing by second-order trigeminal brain stem neurons. GABAA receptor mechanisms act locally at both the Vi/Vc transition and Vc/C1 junction regions to inhibit corneal input and act through polysynaptic pathways to modify corneal input at multiple levels of the trigeminal brain stem complex. PMID- 12890798 TI - Postlesional vestibular reorganization improves the gain but impairs the spatial tuning of the maculo-ocular reflex in frogs. AB - The ramus anterior (RA) of N.VIII was sectioned unilaterally. Two months later we analyzed in vivo responses of the ipsi- and of the contralesional abducens nerve during horizontal and vertical linear acceleration in darkness. The contralesional abducens nerve had become responsive again to linear acceleration either because of a synaptic reorganization in the vestibular nuclei on the operated side and/or because of a reinnervation of the utricular macula by regenerating afferent nerve fibers. Significant differences in the onset latencies and in the acceleration sensitivities allowed a separation of RA frogs in a group without and in a group with functional utricular reinnervation. Most important, the vector orientation for maximal abducens nerve responses was clearly altered: postlesional synaptic reorganization resulted in the emergence of abducens nerve responses to vertical linear acceleration, a response component that was barely detectable in RA frogs with utricular reinnervation and that was absent in controls. The ipsilesional abducens nerve, however, exhibited unaltered responses in either group of RA frogs. The altered spatial tuning properties of contralesional abducens nerve responses are a direct consequence of the postlesional expansion of signals from intact afferent nerve and excitatory commissural fibers onto disfacilitated 2nd-order vestibular neurons on the operated side. These results corroborate the notion that postlesional vestibular reorganization activates a basic neural reaction pattern with more beneficial results at the cellular than at the network level. However, given that the underlying mechanism is activity-related, rehabilitative training after vestibular nerve lesion can be expected to shape the ongoing reorganization. PMID- 12890799 TI - Role of the electrogenic Na/K pump in disinhibition-induced bursting in cultured spinal networks. AB - Disinhibition-induced bursting activity in cultures of fetal rat spinal cord is mainly controlled by intrinsic spiking with subsequent recurrent excitation of the network through glutamate synaptic transmission, and by autoregulation of neuronal excitability. Here we investigated the contribution of the electrogenic Na/K pump to the autoregulation of excitability using extracellular recordings by multielectrode arrays (MEAs) and intracellular whole cell recordings from spinal interneurons. The blockade of the electrogenic Na/K pump by strophanthidin led to an immediate and transient increase in the burst rate together with an increase in the asynchronous background activity. Later, the burst rate decreased to initial values and the bursts became shorter and smaller. In single neurons, we observed an immediate depolarization of the membrane during the interburst intervals concomitant with the rise in burst rate. This depolarization was more pronounced during disinhibition than during control, suggesting that the pump was more active. Later a decrease in burst rate was observed and, in some neurons, a complete cessation of firing. Most of the effects of strophanthidin could be reproduced by high K+-induced depolarization. During prolonged current injections, spinal interneurons exhibited spike frequency adaptation, which remained unaffected by strophanthidin. These results suggest that the electrogenic Na/K pump is responsible for the hyperpolarization and thus for the changes in excitability during the interburst intervals, although not for the spike frequency adaptation during the bursts. PMID- 12890800 TI - Beta-amyloid peptide activates non-alpha7 nicotinic acetylcholine receptors in rat basal forebrain neurons. AB - Alzheimer's disease (AD) is a progressive neurodegenerative condition characterized by profound deficits in memory and cognitive function. Neuropathological hallmarks of the disease include a loss of basal forebrain cholinergic neurons and the deposition of beta-amyloid peptide (Abeta) in neuritic plaques. At a cellular level, considerable attention has focused on a study of Abeta interactions with the neuronal nicotinic acetylcholine receptor (nAChR) subtypes. In this study, using cell-attached and outside-out single channel recordings from acutely dissociated rat basal forebrain neurons, we report that Abeta and nicotine activate nAChRs with two distinct levels of single channel conductance. Whole cell recordings from these neurons reveal Abeta and nicotine, in a concentration-dependent and reversible manner, evoke brisk depolarizing responses and an inward current. The effects of Abeta on both single channel and whole cell are blocked by the noncompetitive nAChR antagonist mecamylamine and competitive nAChR antagonist dihydro-beta-erythroidine, but not the specific alpha7-selective nAChR antagonist methyllycaconitine, indicating that Abeta activated non-alpha7 nAChRs on basal forebrain neurons. In addition, the non-alpha7 nAChR agonists UB-165, epibatidine, and cytisine, but not the selective alpha7 agonist AR-R17779, induced similar responses as Abeta and nicotine. Thus non-alpha7 nAChRs may also represent a novel target in mediating the effects of Abeta in AD. PMID- 12890801 TI - Best Practice No 172: pituitary gland pathology. AB - This article reviews published evidence on the diagnosis and classification of pituitary gland tumours and the relevance of histological and genetic features to prognosis. Much of the literature is devoted to the histological, ultrastructural, and immunocytochemical classification of pituitary adenomas (extensively supported by multicentre studies), with little consensus on the identification of prognostic features in adenomas, particularly in relation to invasion. There is a lack of correspondence between clinical and pathological criteria to identify and classify invasion, and a need to reassess the nomenclature and diagnostic criteria for invasive adenomas and carcinomas. Recent cytogenetic, genetic, and molecular biological studies have identified no consistent abnormalities in relation to pituitary tumour progression, although many genes are likely to be involved. In light of these uncertainties, an approach to the diagnosis and classification of pituitary adenomas is suggested, based on robust criteria from earlier studies and incorporating provisional data that require reassessment in large prospective studies with an adequate clinicopathological database. PMID- 12890804 TI - Increase of bone marrow lymphocytes in systemic mastocytosis: reactive lymphocytosis or malignant lymphoma? Immunohistochemical and molecular findings on routinely processed bone marrow biopsy specimens. AB - AIMS: To clarify the nature (reactive or neoplastic) of lesional, perifocally aggregated lymphocytes in bone marrow infiltrates of systemic mastocytosis (SM), the histopathology of which can resemble malignant lymphoma with focal bone marrow involvement, particularly low grade malignant B cell lymphoma of lymphoplasmacytic immunocytoma subtype, which frequently exhibits increased mast cell (MC) numbers. METHODS: Thirteen cases of SM and three of lymphoplasmacytic immunocytoma with predominant focal bone marrow infiltration were investigated. Immunostaining of formalin fixed, paraffin wax embedded bone marrow specimens was performed using antibodies against CD2, CD5, CD20, CD23, and CD25; kappa and lambda immunoglobulin light chains; and MC markers chymase, tryptase, and CD117 (KIT). Monoclonal rearrangements of IgH and TCRgamma were studied using seminested polymerase chain reaction (PCR). c-kit point mutation Asp816-Val was detected by PNA mediated PCR clamping and hybridisation probes. RESULTS: The lymphocytic clusters in SM contained nearly equal numbers of mature T and B cells, the latter with no coexpression of aberrant antigens, such as CD5 or CD23. Most MCs in SM cases constantly coexpressed tryptase, CD25, and CD117. No monoclonal rearrangements were seen for IgH or TCRgamma. In contrast, B cells from immunocytomas showed light chain restriction and monoclonal rearrangement for IgH, confirming their neoplastic nature. c-kit point mutation Asp816-Val was found in ten of 13 SM cases, but in none of the three immunocytomas. CONCLUSIONS: Focal accumulations of lymphocytes in the bone marrow of SM are reactive in nature and could be termed lymphocytosis. A diagnosis of SM-AHNMD/immunocytoma should not be made. PMID- 12890803 TI - Expression of folate receptors and heterogeneous nuclear ribonucleoprotein E1 in women with human papillomavirus mediated transformation of cervical tissue to cancer. AB - AIMS: Folate receptors (FRs) mediate cellular uptake of folates in many cancer cells and in folate deficiency heterogeneous nuclear ribonucleoprotein E1 (hnRNP E1) mediates translational upregulation of FR in cultured cervical cancer cells. hnRNP-E1 can also interfere with human papillomavirus 16 (HPV-16) viral capsid protein synthesis (and thereby HPV proliferation) in vitro. This study aimed to evaluate prospectively the relevance of FR and hnRNP-E1 expression in the normal cervix, cervical dysplasia, and cancer. METHODS: Cervical tissues from 12 women with normal histology and 69 consecutive women with varying grades of cervical dysplasia and cancer were prospectively evaluated for immunohistochemical expression of FR, hnRNP-E1, proliferating cell nuclear antigen (PCNA), and HPV. There were 22 women with low grade squamous intraepithelial lesions (LGSIL), 22 with high grade squamous intraepithelial lesions (HGSIL), and 25 with invasive cervical carcinoma. RESULTS: Among normal subjects, 100% and 92% expressed hnRNP E1 and FR, respectively. FR expression decreased from 91% in LGSIL to 68% and 64% in women with HGSIL and cancer, respectively. Similarly, hnRNP-E1 expression decreased from 86% in LGSIL to 68% and 40% in HGSIL and cancer, respectively. There was a highly significant positive correlation between the extent of FR and hnRNP-E1 expression, and an inverse correlation between HPV infection and hnRNP E1 expression during progression of cervical dysplasia to cancer. CONCLUSION: These results are consistent with a hypothesis that reduced hnRNP-E1 expression may be permissive for HPV proliferation and progression to cervical cancer, and support the need for prospective longitudinal studies of hnRNP-E1 expression in HPV-16 infected women. PMID- 12890805 TI - Prognostic significance of grading (MIB-1 system) in patients with myxoid liposarcoma. AB - AIMS: To determine the relation between clinical outcome and tumour grade defined by a MIB-1 (Ki-67) score based grading system. METHOD: The clinical and pathological features of 50 patients with myxoid liposarcoma were evaluated, and MIB-1 immunostaining was performed to grade these patients' tumours. Univariate and multivariate analyses were conducted to evaluate survival. Clinical follow up details were available for all patients (median, 46.5 months; range, 9-408). RESULTS: Univariate analysis revealed that the tumour site (p < 0.05), round cell component content (p < 0.01), necrosis (p < 0.01), mitosis (p < 0.01), MIB-1 labelling index (p < 0.001), and tumour grade (p < 0.001) had a significant impact on overall survival. Multivariate analysis showed that, of the variables evaluated, the tumour grade defined by a MIB-1 score based grading system was the most significant adverse prognostic factor. CONCLUSION: Tumour grade determined by the grading system using the MIB-1 score (MIB-1 system) is a very strong prognostic factor in patients with myxoid liposarcoma. PMID- 12890806 TI - Enhanced telomere shortening in transformed lymphoblasts from patients with X linked dyskeratosis. AB - AIM: Dyskeratosis congenita (DC) is characterised by the failure of those tissues that are rapidly dividing in the adult, particularly the skin, mucosae, and haemopoietic system. The X linked form of the disease is caused by mutations of the DKC1 gene, which encodes dyskerin, a protein that is necessary for the function of telomerase. Cultured DC lymphoblastoid cells are characterised by a reduced expansion of the cell population because of the progressive increase in apoptosis compared with the number of cell divisions. This report aimed to verify whether this is caused by a defect in telomerase function. METHODS: Variations in telomere length over time were evaluated in two cultured lymphoblastoid cell lines derived from patients with X linked DC and control cells derived from a non affected individual. In addition, the effect of inhibiting poly (ADP-ribose) polymerase (PARP), which is involved in the cellular response to excessive telomere shortening, was assessed. One DC cell line and the control cells were treated with the specific PARP inhibitor 1,5-dihydroxyquinoline (IQ). RESULTS: In DC cells the increase in cell death was associated with progressive telomere shortening, and this was not seen in the control cells. Treatment with IQ delayed the increase of apoptosis in DC cells. CONCLUSIONS: These observations indicate that the reduced expansion that characterises cultured cells obtained from patients with X linked DC is caused by premature telomere shortening. PMID- 12890807 TI - p27 Kip1 protein expression in Hashimoto's thyroiditis. AB - AIMS: Hashimoto's thyroiditis (HT) is an autoimmune disease in which both proliferation and apoptosis are enhanced. p27(Kip1) protein protects tissues from disease mechanisms that involve excessive cell proliferation and apoptosis. This study investigated whether there is loss of p27(Kip1) expression in HT and whether p27(Kip1) immunoreactivity has any relation to the proliferative indicator Ki-67. Because p27(Kip1) is regulated through either degradation, mediated by the S phase kinase associated protein 2 (Skp2), or sequestration, via D3 cyclin, the expression of these proteins was also investigated. METHODS: Immunohistochemistry was used to assess p27(Kip1), Ki-67, Skp2, and cyclin D3 expression in 19 cases of HT and in 10 normal thyroids. The results were evaluated by image analysis and reported as labelling indices (LIs) in both groups. RESULTS: The p27(Kip1) LI was lower in HT than in normal thyroid (28% v 75%; p < 0.001), whereas Ki-67 (1.13% v 0.13%), Skp2 (0.74% v 0.15%), and cyclin D3 (1.56% v 0.00%) LIs were higher in HT than in normal thyroids (p < 0.001). There was no correlation between p27(Kip1) and the expression of Ki-67, Skp2, and cyclin D3. CONCLUSIONS: p27(Kip1) downregulation is not exclusive to tumours but occurs also in HT, independently of the proliferative status and of changes in Skp2 and cyclin D3 expression. Further investigation is required to understand the mechanisms leading to p27 deregulation because these observations suggest that the regulation of p27(Kip1) expression in epithelial thyroid cells may play a role in HT pathogenesis. PMID- 12890808 TI - Outcome of women with inadequate cervical smears followed up for five years. AB - BACKGROUND: The clinical and prognostic significance of "inadequate" cervical smear is unknown, even though women with repeated inadequate smears are referred for colposcopy in the National Health Service (NHS) Cervical Screening Programme. AIM: To follow up a cohort of women with inadequate cervical smears over the following five years to examine outcomes, including detection of high grade cervical intraepithelial neoplasia (CIN). METHODS: The study comprised 1972 women with an inadequate cervical smear reported at Walsall Hospitals NHS Trust between 1 April 1995 and 31 March 1996. Results of cervical smears and biopsies taken over the following five years were reviewed to confirm the outcome. FINDINGS: Within five years, 2.2% of women with an inadequate cervical smear developed histologically confirmed high grade CIN, which was higher than the 1.3% seen among all women with cervical smear tests reported at the same laboratory over the same period, although the difference was not significant at the 95% level of confidence. Where inadequacy resulted from or was contributed to by "polymorphs obscuring", the risk of subsequent development/detection of high grade CIN was 2.6%. CONCLUSIONS: Women with inadequate cervical smears had an increased risk of detection of high grade CIN in the following five years compared with "all women". This increased risk was not significant, although if a larger number of women had been studied significance may have been reached, so that further studies are needed. The increased risk appeared to be at least partially dependent on the reason for inadequacy. PMID- 12890809 TI - Unconjugated bilirubin in human bile: the nucleating factor in cholesterol cholelithiasis? AB - AIMS: To investigate the concentrations of bilirubin, bilirubin conjugates, phospholipid, and cholesterol in the gall bladder bile obtained at surgery from patients with and without cholesterol gallstones. METHODS: Gall bladder bile was collected during surgery, by puncture, from 20 patients with gallstones undergoing routine cholecystectomy and from eight patients with normal liver blood tests. Concentrations of bilirubin, bilirubin conjugates, phospholipid, and cholesterol were measured using standard procedures. RESULTS: The proportion of total bilirubin that was unconjugated was significantly higher in the bile from patients with stones than in bile from control patients, whether or not the bile from either group was saturated with cholesterol or not. Indeed, the mean concentration of cholesterol was significantly higher in control bile samples. CONCLUSION: The presence of stones was more closely related to the proportion of unconjugated bilirubin than to the degree of saturation of bile with cholesterol. Bilirubin and its metabolites probably play an important part in the formation of cholesterol gallstones. PMID- 12890810 TI - Insulin-like growth factor system gene expression in women with type 2 diabetes and breast cancer. AB - BACKGROUND/AIMS: A twofold increased risk for breast cancer has been reported recently for women with late onset diabetes. Most studies showed that there were differences in serum concentrations of insulin-like growth factors and related proteins between women with and without diabetes who have breast cancer. This study investigated the expression of these markers at the cellular level in a cohort of women with and without type 2 diabetes who underwent biopsy because of a breast lump. METHODS: Relative quantitative analysis of specific mRNA sequences was performed after extraction and reverse transcription polymerase chain reaction amplification from formalin fixed and paraffin wax embedded tissues. Sixty seven breast surgical specimens from women with and without diabetes who did not have cancer and from women with and without diabetes who did have cancer were studied for insulin-like growth factor I (IGF-I), the IGF-I receptor (IGF IR), insulin-like growth factor binding protein 3 (IGFBP-3), and oestrogen receptor 1 gene expression. RESULTS: The expression of IGF-I and IGF-IR was significantly lower in the cancer groups, whereas there was no significant difference for IGFBP-3 between women with and without cancer. Moreover, there was a good correlation between the expression of IGF-I and IGF-IR in women without cancer: this link was still present in breast tissue from patients with diabetes and cancer, whereas it was lost in patients without diabetes but with cancer. CONCLUSIONS: These differences in IGF-I/IGF-IR expression could contribute to the increased risk for breast cancer in women with type 2 diabetes. PMID- 12890811 TI - External quality assessment for warfarin dosing using computerised decision support software. AB - AIM: To establish and evaluate an external quality assessment scheme for warfarin dosing for users of a computerised decision support system, BAP-PC. DESIGN: Analysis of 12 months of clinical data from 10 primary care centres using BAP-PC within an oral anticoagulation clinic. Data were analysed for individual centres and compared with aggregated data for all practices. Individual feedback forms were provided to participating centres. RESULTS: A total patient population of 367 (range, 17-65/centre) was analysed. On average, patients spent 69% of time in the therapeutic range (range, 60-76%). Patients were seen on average every 27 days (range, 24-30). The average point prevalence was 86% (range, 76-100%). In total, 33 adverse events were reported (0-13/practice). Serious adverse events ranged from 0 to 1 for each practice. This translates into a serious adverse event rate of 1.6/100 patient years. CONCLUSIONS: Practices were successful in maintaining good therapeutic international normalised ratio control, with centres achieving 60% or higher time in range. There are some doubts about the quality of data collection at a practice level because there were no reported events in half of the participating centres. The observed event rates do concur with previously reported data, however. Further cycles of the scheme are necessary to establish it as a useful research and benchmarking tool. PMID- 12890812 TI - A comparison of the performance of commercially available chromogenic agars for the isolation and presumptive identification of organisms from urine. AB - AIMS: To compare four media-UTI medium, BBL CHROMagar, CPS ID2, and Harlequin CLED-using a collection of fully characterised organisms and subsequent "field trial". METHODS: Seven hundred and eighty seven fully characterised isolates (730 Gram negative bacteria, 47 Gram positive bacteria, and 10 yeasts) were used to test for accuracy of organism identification. To assess isolation rates and ability to detect mixed cultures, 1435 urine samples were cultured in the three best performing chromogenic media (UTI medium, BBL CHROMagar, and CPS ID2) and CLED. RESULTS: The chromogenic agars differed in their accuracy of identification, with BBL CHROMagar performing best and Harlequin CLED performing least well. Similarly, BBL CHROMagar achieved a higher overall isolation rate than UTI medium and CPS ID2. When mixed growth was defined as greater than two organism types, BBL CHROMagar detected more mixed cultures than did UTI medium and CPS ID2, although the differences were not significant. When mixed growth was defined as greater than one organism type the increased number of mixed growths detected by BBL CHROMagar became significant, largely because of differences in enterococcal isolation rates. CONCLUSION: The use of BBL CHROMagar, UTI medium, or CPS ID2 chromogenic agar as a replacement for CLED agar would improve the detection rate of contaminated urine samples. Enhanced identification helps to distinguish different species, facilitating the monitoring of bacterial resistance in support of the national antibiotic strategy. BBL CHROMagar gave the highest overall organism recovery rates, greatest ability to detect mixed cultures, and the most accurate identification of organisms. PMID- 12890813 TI - Optimisation of acid fast smears for the direct detection of mycobacteria in clinical samples. AB - AIMS: Despite its long history, the acid fast smear remains unstandardised. Technical variations in both the preparation of clinical material and subsequent staining mean that smear sensitivity relative to culture may vary from 50% to over 80%. This study assessed the sensitivity of acid fast microscopy at each of five stages of sample preparation and by both commonly used staining methods. METHODS: Sputum samples thought for varying reasons to be highly likely to be culture positive were used to prepare a series of smears in which the effects of digestion (liquefaction), concentration (centrifugation), and decontamination (sodium hydroxide) could be assessed, together with a comparison of staining by the auramine/phenol and Ziehl-Neelsen techniques. RESULTS: The most effective method for the demonstration of acid fast organisms in sputum was found to be an auramine phenol stain applied to a liquefied, concentrated sample and examined before the decontamination process. CONCLUSIONS: The auramine phenol stain applied to a liquefied, concentrated sample and examined before the decontamination process is the most effective method for the demonstration of acid fast organisms in sputum. PMID- 12890814 TI - Associations between Mycoplasma genitalium, Chlamydia trachomatis and pelvic inflammatory disease. AB - OBJECTIVE: To evaluate the association between Mycoplasma genitalium, Chlamydia trachomatis, and pelvic inflammatory disease (PID) METHODS: A case-control methodology was used. Swab eluates were processed using the QIAamp DNA mini kit. Polymerase chain reaction (PCR) for M genitalium was carried out using a real time in-house 16S based assay. An endocervical swab was taken and tested for the presence of C trachomatis (ligase chain reaction, Abbott Laboratories), and a high vaginal swab was taken and tested for the presence of Neisseria gonorrhoeae and bacterial vaginosis. RESULTS: Of the PID cases 13% (6/45) had evidence of M genitalium infection compared to none of the controls (0/37); 27% (12/45) of the cases had C trachomatis infection compared to none of the controls; and 16% (7/45) of cases only had serological evidence of C trachomatis infection compared to 5% (2/37) of controls. Cases were more likely to present with M genitalium and/or C trachomatis than controls (p<0.001). CONCLUSIONS: This study indicates that there may be an association between M genitalium and PID, and that this relation is largely independent of C trachomatis. Future studies need to investigate the pathological basis of the relation between M genitalium and PID using samples from women with PID diagnosed using laparoscopy and endometrial biopsy. Little is known about the epidemiology of M genitalium: large scale epidemiological investigations are needed to determine the prevalence, incidence, and factors associated with this emerging infection. PMID- 12890815 TI - Correction of uneven illumination (vignetting) in digital microscopy images. AB - BACKGROUND: Many digital microscopy images suffer from poor illumination at the peripheries (vignetting), often attributable to factors related to the light path between the camera and the microscope. A commonly used method for correcting this has been to use the illumination of an empty field as a correction filter (white shading correction). AIMS: To develop an alternative shading correction method that does not require this additional image. METHODS: / RESULTS: This report outlines an alternative shading correction method that is based upon the intrinsic properties of the image, which are revealed through Gaussian smoothing. The technique can be implemented within Adobe Photoshop or other graphics editing software and works well with both haematoxylin and eosin and immunohistochemical images. CONCLUSIONS: This technique provides an effective means of optimising digital microscopy image appearances for printing, image analysis, or telepathology. PMID- 12890816 TI - Measurement of 5-hydroxy-2-aminovaleric acid as a specific marker of metal catalysed oxidation of proline and arginine residues of low density lipoprotein apolipoprotein B-100 in human atherosclerotic lesions. AB - gamma-Glutamyl-semialdehyde (gammaGSA) is a major product of the metal catalysed oxidation of apolipoprotein B-100 (apoB-100) proline and arginine residues. On reduction, gammaGSA forms 5-hydroxy-2-aminovaleric acid (HAVA). This report describes the application of HAVA measurement to characterise the formation of gammaGSA in low density lipoprotein (LDL) recovered from human atherosclerotic lesions. HAVA concentrations were greatly increased in LDL from early (mean, 10.25; SD, 3.49 mol/mol apoB-100; p < 0.01), intermediate (mean, 11.18; SD, 2.37 mol/mol apoB-100; p < 0.01), and advanced (mean, 9.91; SD, 2.15 mol/mol apoB-100; p < 0.01) lesions, when compared with LDL from normal aortic tissue (mean, 0.05; SD, 0.01 mol/mol apoB-100). These findings support the hypothesis that pathways involving metal catalysed oxidation of LDL apoB-100 are of pathological importance in atherogenesis. PMID- 12890817 TI - The effect of calpain 3 deficiency on the pattern of muscle degeneration in the earliest stages of LGMD2A. AB - Limb girdle muscular dystrophy type 2A (LGMD2A) is caused by mutations in the calpain 3 gene. In a large family affected by LGMD2A with four severely affected members, three additional asymptomatic relatives had very high serum creatine kinase concentrations. All were homozygous for the R110X mutation and showed a total absence of calpain 3 in the muscle. Histological analysis of muscle in these three rare preclinical cases showed a consistent but unusual pattern, with isolated fascicles of degenerating fibres in an almost normal muscle. This pattern was also seen in one patient with early stage LGMD2A who had a P82L missense mutation and a partial deficiency of calpain 3 in the muscle, but was not seen in early stage patients affected by other forms of LGMD. These findings suggest that a peculiar pattern of focal degeneration occurs in calpainopathy, independently of the type of mutation or the amount of calpain 3 in the muscle. PMID- 12890819 TI - Late pulmonary metastasis in uterine PEComa. AB - Perivascular epithelioid cell tumours (PEComas) other than angiomyolipoma, clear cell "sugar" tumour of the lung, and lymphangioleiomyomatosis are very rare mesenchymal tumours. The uterus seems to be the most prevalent site of involvement, but only 13 cases of uterine PEComa have been described. Three of these cases exhibited local aggressive behaviour and only one showed metastasis. Because of the extremely small number of cases, PEComas are considered tumours of uncertain malignant potential. This report describes a 68 year old woman, who presented with multiple pulmonary lesions seven years after the initial diagnosis of well differentiated endometrial stromal sarcoma. Histological and immunohistochemical analysis of the pulmonary lesions, in addition to re evaluation of the primary uterine tumour, led to the final diagnosis of metastatic uterine PEComa. The findings indicate that any PEComa might have malignant potential. Spreading of this tumour to other organs might become evident even several years after primary manifestation. PMID- 12890820 TI - Intestinal goblet cell autoantibody associated enteropathy. AB - This report describes a case of refractory enteropathy with circulating intestinal goblet cell autoantibodies (IGA). A 19 year old man with hyperthyroidism had suffered from protracted diarrhoea for nearly 10 years. Histological examination showed evidence of collagenous enterocolitis. The diarrhoea did not improve despite fasting under total parenteral nutrition. An immunofluorescence assay demonstrated IGA without anti-enterocyte autoantibodies, the hallmark of autoimmune enteropathy, although other criteria were fulfilled. None of 109 controls, including 55 cases of inflammatory bowel disease and one of lymphocytic colitis, had IGA. This case is considered to be a variant of autoimmune enteropathy, and might be a distinct entity. PMID- 12890821 TI - Hepatosplenic T cell lymphoma with no expression of cytotoxic molecules. AB - Hepatosplenic T cell lymphoma is defined as an extranodal and systemic neoplasm derived from cytotoxic T cells. This report describes a postmortem case of T cell lymphoma that showed histological features of hepatosplenic T cell lymphoma but did not express cytotoxic molecules. The patient was a 57 year old man who presented with severe icterus and hepatosplenomegaly, followed by an aggressive clinical course. The liver and spleen were enlarged, weighing 2000 g and 360 g, respectively. Histologically, the liver, spleen, and bone marrow were entirely affected by lymphoma, comprising pleomorphic small and large cells, which displayed sinusoidal infiltration in the liver, diffuse infiltration in the splenic cord, and interstitial/diffuse infiltration with fibrosis in the bone marrow. Lymphoma cells showed positivity for CD3 epsilon, CD8, and CD45RO and clonal rearrangement of the TCRgamma gene by the polymerase chain reaction on paraffin wax embedded sections. However, they were negative for TIA-1 and granzyme B, in addition to betaF1, CD4, and CD56. Few neoplastic cells were stained for Epstein-Barr virus encoded mRNA 1. These findings indicate that this case might represent a variant of hepatosplenic T cell lymphoma despite the absence of cytotoxic molecules. PMID- 12890822 TI - Primary venous dissecting aneurysm arising during pregnancy: a case report and review of the literature. AB - This report describes a case of venous dissecting aneurysm presenting as a popliteal mass, in a 33 year old woman. A 1 x 1 x 0.5 cm lump developed early in the course of a second pregnancy. Primary venous aneurysms are rare vascular abnormalities that can affect either the superficial or deep veins, and have been described throughout the venous system. Most commonly found in the neck and central thoracic veins, they have also been found in visceral veins and extremities. There is a tendency for vascular disturbances to occur during pregnancy. The haemodynamic changes and hormonal milieu may be the cause of vascular alterations, which can lead to new aneurysm formation, or weakening of pre-existing aneurysms. This is the first reported case of a dissecting venous aneurysm, and has the added interest that it occurred during pregnancy. PMID- 12890823 TI - Fungus colonisation of pulmonary rheumatoid nodule. AB - This report describes a 69 year old woman, suffering from active rheumatoid arthritis since the age of 60 and presenting with severe dyspnoea and cough. A computed tomography scan of the chest showed multiple bilateral pulmonary nodules, sometimes cavitated, associated with reticular opacities and pleural effusion. A videothoracoscopic excision of a cavitated nodule was performed. Seven days after surgery, a right pneumothorax developed, and the patient died of septicaemia one month later. Microscopically, the excised nodule was composed of necrotic fibrinoid material with a peripheral rim of palisaded histiocytes, extending to the pleural surface and containing several fungal hyphae morphologically consistent with aspergillus. A diagnosis of pulmonary rheumatoid nodule with fungus colonisation was made. In the lung, fungus colonisation is a rare complication of rheumatoid nodules. The most important differential diagnostic considerations are briefly discussed. PMID- 12890824 TI - Caution should be taken in using CD31 for distinguishing the vasculature of lymph nodes. PMID- 12890825 TI - An early lesion of pulmonary carcinosarcoma: possible diagnostic problem in frozen section interpretation. PMID- 12890826 TI - Cirrhosis with steatohepatitis following longterm stilboestrol treatment. PMID- 12890827 TI - 'The hand-arm vibration syndrome'--a prevention challenge or a price to pay? PMID- 12890828 TI - A medical assessment process for a large volume of medico-legal compensation claims for hand-arm vibration syndrome. AB - BACKGROUND: The judgement on preliminary issues of nine lead cases of suspected hand-arm vibration syndrome (HAVS) in former coal miners in the UK ruled that there was evidence of damage and breach of duty of care (Armstrong and Others v. British Coal, 1996). In anticipation of > 120 000 cases and at significant cost, a handling agreement was prepared in 1999. This recommended a single medical assessment process (MAP) to determine general damages for which almost 200 doctors attended 2 days of training. AIMS: This paper outlines the assessment process and the results to date. METHODS: Seventeen test centres across the UK were commissioned. Standardization of the performance by both the doctors and technicians within individual examination centres and across all centres was necessary. A pragmatic solution using 'best available assessment techniques', whilst at the same time coping with the large number of claimants, was required. Doctors were trained to administer questionnaires for clinical symptoms, past medical history and occupational history, and a standardized clinical assessment pro-forma. Three standardized tests were used: vibrotactile thresholds, thermal aesthesiometry and cold water provocation testing. A modification of the Stockholm Workshop Scales and scoring system was adopted. At time of writing, 52 490 claimants had been assessed by the MAP. RESULTS: Analysis of results showed that 5% were assessed at 0SN, 15% at 1SN, 18% at 2SN (early), 28% at 2SN (late), 33% at 3SN, 21% at 0V, 13% at 1V, 38% at 2V and 28% at 3V. CONCLUSION: It is concluded that the MAP is a practical and time-efficient tool for assessing a large volume of claimants with suspected HAVS. Further analysis of the process and staging is required to confirm its validity as a medico-legal examination. PMID- 12890829 TI - The objective diagnosis of early tennis elbow by magnetic resonance imaging. AB - OBJECTIVE: To identify the salient magnetic resonance imaging (MRI) features of tennis elbow. An objective diagnosis is important when managing work-related incapacity due to ill-defined lateral arm pain. METHOD: Twenty-three symptomatic and 17 asymptomatic elbows in 20 patients with tennis elbow, no evidence of other pathology and no previous treatment were imaged using established MRI sequences. RESULTS: In the symptomatic elbows, the common extensor origin (CEO) showed signs of oedema in 23, thickening in 19, peri-tendon oedema in 3 and tears in 13 cases. More extensive abnormalities were demonstrated in only two elbows. Six out of 17 asymptomatic elbows also showed oedema in the CEO. CONCLUSIONS: The CEO is confirmed as the primary site of MRI changes in tennis elbow. Oedema was commonly found in asymptomatic elbows, necessitating the presence of thickening or tears in the CEO tendon to objectively diagnose tennis elbow on MRI. PMID- 12890830 TI - Towards epidemiological criteria for soft-tissue disorders of the arm. AB - BACKGROUND: The lack of universally agreed criteria has hampered population studies of the prevalence and causation of soft-tissue disorders of the upper limb. OBJECTIVES: To establish core variables for classification of the commonest disorders seen in population samples. METHODS: Consecutive new cases seen in clinical practice in five different centres were evaluated with respect to 30 variables shown to have discriminatory value in univariate analysis. Multivariate analysis using logistic regression modelling was carried out with these as the independent variables and with the clinical diagnosis as the dependent variable. RESULTS: A total of 1382 cases of soft-tissue disorder were recorded and only those diagnostic groups with 50 or more cases were included. In multivariate logistic regression, significant variables positively discriminating for each disorder were identified for carpal tunnel syndrome (n = 56), lateral epicondylitis (n = 87), tenosynovitis (n = 63), shoulder tendonitis (n = 157), non-specific upper limb disorder (n = 458), fibromyalgia (n = 124) and inflammatory arthritis (n = 100), which was used for comparison purposes. Significant discrimination for each model was demonstrated by the construction of receiver operating characteristic (ROC) curves and appropriate area under the curve statistics. CONCLUSIONS: This approach to classification criteria is based on multivariate modelling rather than on a consensus statement. This includes the effects of negative as well as positive associations. Further work is required on both the reproducibility of the clinical signs and the application of the criteria to other datasets. PMID- 12890831 TI - Hypothenar hammer syndrome: a discrete syndrome to be distinguished from hand-arm vibration syndrome. AB - BACKGROUND: Hypothenar hammer syndrome (HHS) is a cause of vascular insufficiency to the hand and may be manifest as Raynaud's phenomenon. The cause is trauma to the vulnerable portion of the ulnar artery as it passes over the hamate bone, which may result in thrombosis, irregularity or aneurysm formation. AIM: This review was undertaken in order to clarify the features of HHS that may differentiate it from hand-arm vibration syndrome. METHODS: A tiered review of world literature was undertaken using Medline and EMBase as the primary search engines. Fifty-two relevant articles were critically reviewed. CONCLUSION: Colour and temperature changes occur more diffusely in HHS than in classical Raynaud's phenomenon and the absence of the triphasic colour change may alert clinicians to the diagnosis, which may be confirmed by Allen's test. Doppler or arteriographic studies are required for confirmation. It is important to recognize the possibility of HHS in the occupational setting as a potentially curable cause of Raynaud's phenomenon, distinct from hand-arm vibration syndrome. The possibility exists of HHS occurring as a result of repeated hypothenar trauma from vibrating tools, in which case the nature and magnitude of the individual episode of trauma may be more important than the weighted acceleration level of vibration exposure. PMID- 12890832 TI - A critique of a UK standardized test of finger rewarming after cold provocation in the diagnosis and staging of hand-arm vibration syndrome. AB - BACKGROUND: Accurate diagnosis and staging of hand-arm vibration syndrome (HAVS) is important in health surveillance of vibration-exposed workers and the substantial number of related medico-legal cases. The measurement of the rewarming rate of fingers after cold provocation to the hands (CPT) has been suggested as a useful test in diagnosing HAVS. AIM: To investigate the diagnostic value of a standardized version of the CPT test using a 15 degrees C cold challenge for 5 min applied in the recent compensation assessment of UK miners. METHODS: Analysis of a subset of UK miners assessed at our unit, together with data from a small repeatability study of the standardized CPT in normal subjects. RESULTS: Rewarming time in the CPT was significantly lower in those subjects classified as vascular Stockholm stage 0 compared with Stockholm stages 1-3 combined, but did not discriminate between the stages of abnormality. Using the suggested cut-off in the CPT test, the sensitivity and specificity were calculated as 43 and 78%, respectively. Receiver operator characteristic analysis suggested that the rewarming time of highest accuracy gave a sensitivity of 66% and specificity of 59%. In 10 miners who reported unilateral hand blanching, there was no significant difference in CPT measurements between blanching and non blanching hands. Repeat CPT measurements in normal subjects suggested mean differences of 52 and 107 s for each hand, and the Bland-Altman coefficient of repeatability was approximately 600 s for all fingers. CONCLUSION: Single application of this standardized CPT test may have limited value in diagnosing the vascular component of HAVS in an individual. PMID- 12890833 TI - Thoracic outlet syndrome--aspects of diagnosis in the differential diagnosis of hand-arm vibration syndrome. AB - BACKGROUND: Thoracic outlet syndrome (TOS) is a cause of vascular and neurological compromise to the arm and hand, and may manifest as Raynaud's phenomenon. It may be under-diagnosed. AIM: This review was undertaken in order to clarify the diagnostic and investigative features of TOS that may differentiate it from hand-arm vibration syndrome. METHODS: A tiered review of the world literature was undertaken using Medline and Embase as the primary search engines. CONCLUSION: Thoracic outlet syndrome most commonly presents with neurological symptoms in the arm. Vascular symptoms, including Raynaud's phenomenon, may occur in approximately 10% of cases. Careful clinical assessment by history and examination may reveal the elements of forearm and upper arm symptoms with postural exacerbation, which distinguish this condition from hand arm vibration syndrome. The usefulness of investigation is unclear, but Doppler and neuroelectric studies may be valuable. Magnetic resonance imaging scan is the investigation of choice. PMID- 12890834 TI - The clinical assessment of hand-arm vibration syndrome. PMID- 12890835 TI - Raynaud's phenomenon in a pneumatic tool worker. 1953. PMID- 12890836 TI - Comments on Coughlin et al.'s paper regarding cold thermography in the objective diagnosis of the hand-arm vibration syndrome. PMID- 12890837 TI - Monitor: effect of lifestyle on health. PMID- 12890838 TI - The von Hippel-Lindau protein, vascular endothelial growth factor, and kidney cancer. PMID- 12890839 TI - Cochlear implants. PMID- 12890840 TI - Do bisphosphonates make children's bones better or brittle? PMID- 12890841 TI - A randomized trial of bevacizumab, an anti-vascular endothelial growth factor antibody, for metastatic renal cancer. AB - BACKGROUND: Mutations in the tumor-suppressor gene VHL cause oversecretion of vascular endothelial growth factor by clear-cell renal carcinomas. We conducted a clinical trial to evaluate bevacizumab, a neutralizing antibody against vascular endothelial growth factor, in patients with metastatic renal-cell carcinoma. METHODS: A randomized, double-blind, phase 2 trial was conducted comparing placebo with bevacizumab at doses of 3 and 10 mg per kilogram of body weight, given every two weeks; the time to progression of disease and the response rate were primary end points. Crossover from placebo to antibody treatment was allowed, and survival was a secondary end point. RESULTS: Minimal toxic effects were seen, with hypertension and asymptomatic proteinuria predominating. The trial was stopped after the interim analysis met the criteria for early stopping. With 116 patients randomly assigned to treatment groups (40 to placebo, 37 to low dose antibody, and 39 to high-dose antibody), there was a significant prolongation of the time to progression of disease in the high-dose--antibody group as compared with the placebo group (hazard ratio, 2.55; P<0.001). There was a small difference, of borderline significance, between the time to progression of disease in the low-dose--antibody group and that in the placebo group (hazard ratio, 1.26; P=0.053). The probability of being progression-free for patients given high-dose antibody, low-dose--antibody, and placebo was 64 percent, 39 percent, and 20 percent, respectively, at four months and 30 percent, 14 percent, and 5 percent at eight months. At the last analysis, there were no significant differences in overall survival between groups (P>0.20 for all comparisons). CONCLUSIONS: Bevacizumab can significantly prolong the time to progression of disease in patients with metastatic renal-cell cancer. PMID- 12890842 TI - Risk of bacterial meningitis in children with cochlear implants. AB - BACKGROUND: In June 2002, the Food and Drug Administration received reports of bacterial meningitis in patients with cochlear implants for treatment of hearing loss. Implants that included a positioner (a wedge inserted next to the implanted electrode to facilitate transmission of the electrical signal by pushing the electrode against the medial wall of the cochlea) were voluntarily recalled in the United States in July 2002. METHODS: We identified patients with meningitis and conducted a cohort study and a nested case-control investigation involving 4264 children who had received cochlear implants in the United States between January 1, 1997, and August 6, 2002, and who were less than six years of age when they received the implants. We calculated the incidence of meningitis in the cohort and assessed risk factors for meningitis among patients and among 199 controls, using data from interviews with parents and abstracted from medical records. RESULTS: We identified 26 children with bacterial meningitis. The incidence of meningitis caused by Streptococcus pneumoniae was 138.2 cases per 100,000 person-years--more than 30 times the incidence in a cohort of the same age in the general U.S. population. Postimplantation bacterial meningitis was strongly associated with the use of an implant with a positioner (odds ratio, 4.5 [95 percent confidence interval, 1.3 to 17.9], with adjustment for medical, surgical, and environmental factors) and with the joint presence of radiographic evidence of a malformation of the inner ear and a cerebrospinal fluid leak (adjusted odds ratio, 9.3 [95 percent confidence interval, 1.2 to 94.5]). The incidence of meningitis among patients who had received an implant with a positioner remained higher than the incidence among those whose implants did not have a positioner for the duration of follow-up (24 months from the time of implantation). CONCLUSIONS: Parents and health care providers should ensure that all children who receive cochlear implants are appropriately vaccinated and are then monitored and treated promptly for any bacterial infections after receiving the implant. PMID- 12890843 TI - Survival of patients undergoing hemodialysis with paricalcitol or calcitriol therapy. AB - BACKGROUND: Elevated calcium and phosphorus levels after therapy with injectable vitamin D for secondary hyperparathyroidism may accelerate vascular disease and hasten death in patients undergoing long-term hemodialysis. Paricalcitol, a new vitamin D analogue, appears to lessen the elevations in serum calcium and phosphorus levels, as compared with calcitriol, the standard form of injectable vitamin D. METHODS: We conducted a historical cohort study to compare the 36 month survival rate among patients undergoing long-term hemodialysis who started to receive treatment with paricalcitol (29,021 patients) or calcitriol (38,378 patients) between 1999 and 2001. Crude and adjusted survival rates were calculated and stratified analyses were performed. A subgroup of 16,483 patients who switched regimens was also evaluated. RESULTS: The mortality rate among patients receiving paricalcitol was 3417 per 19,031 person-years (0.180 per person-year), as compared with 6805 per 30,471 person-years (0.223 per person year) among those receiving calcitriol (P<0.001). The difference in survival was significant at 12 months and increased with time (P<0.001). In the adjusted analysis, the mortality rate was 16 percent lower (95 percent confidence interval, 10 to 21 percent) among paricalcitol-treated patients than among calcitriol-treated patients. A significant survival benefit was evident in 28 of 42 strata examined, and in no stratum was calcitriol favored. At 12 months, calcium and phosphorus levels had increased by 6.7 and 11.9 percent, respectively, in the paricalcitol group, as compared with 8.2 and 13.9 percent, respectively, in the calcitriol group (P<0.001). The two-year survival rate among patients who switched from calcitriol to paricalcitol was 73 percent, as compared with 64 percent among those who switched from paricalcitol to calcitriol (P=0.04). CONCLUSIONS: Patients who receive paricalcitol while undergoing long term hemodialysis appear to have a significant survival advantage over those who receive calcitriol. A prospective, randomized study is critical to confirm these findings. PMID- 12890844 TI - Bisphosphonate-induced osteopetrosis. PMID- 12890845 TI - Images in clinical medicine. Massive calcification in pseudohypoparathyroidism. PMID- 12890846 TI - Clinical practice. Selecting asymptomatic patients for coronary computed tomography or electrocardiographic exercise testing. PMID- 12890847 TI - Drug-induced hepatotoxicity. PMID- 12890848 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 24-2003. A 10-year-old girl with recurrent bouts of abdominal pain. PMID- 12890849 TI - Paricalcitol as compared with calcitriol in patients undergoing hemodialysis. PMID- 12890850 TI - Intellectual impairment and blood lead levels. PMID- 12890851 TI - Obesity and cancer. PMID- 12890852 TI - Fibroblast growth factor 23 in oncogenic osteomalacia and X-linked hypophosphatemia. PMID- 12890853 TI - ZAP-70 in chronic lymphocytic leukemia. PMID- 12890854 TI - Fatal aspergillosis in a patient with SARS who was treated with corticosteroids. PMID- 12890855 TI - Profile of specific antibodies to the SARS-associated coronavirus. PMID- 12890856 TI - Images in clinical medicine. Carotid-artery thrombosis secondary to basal skull fracture. PMID- 12890857 TI - Mouse and human cells versus oxygen. AB - Mice and humans are at opposite ends of the mammalian spectrum of longevity. A major question in biology is whether this difference can be accounted for by differences in the properties of cells from these two species. A new publication from Judith Campisi's lab reports that human cells in culture are more resistant than mouse cells to the damaging effects of 20% oxygen. The greater burden of DNA damage sustained by mouse cells causes them to rapidly enter a phase of culture in which most cells enter permanent growth arrest (replicative senescence). However, some mouse cells usually escape from senescence and then grow into an immortal cell line. This never happens in human fibroblast cell cultures. Human cells also eventually enter replicative senescence in culture, but this phenomenon is caused by shortening of telomeres and not by DNA damage of the type responsible for mouse cell senescence. Human fibroblasts never spontaneously escape from senescence. This Perspective reviews differences between mouse and human cells that could account for these differences in behavior. Some evidence indicates that human cells are generally more resistant than mouse cells to oxidative damage to DNA, but more needs to be done to confirm this finding and to understand the underlying mechanisms. Whether or not there are differences in the amount of DNA damage caused by oxygen or in the early phase of repair, there may be important differences in the later consequences of DNA damage. Mouse cells appear to be able to continue to divide with DNA damage that has not been repaired or has been misrepaired, and becomes fixed in the form of chromosomal abnormalities. The checkpoints that cause cells to stop dividing when chromosomes develop abnormalities (aberrations or shortened telomeres) appear to operate more efficiently in human cells. Much more work is needed to understand the basis for these differences and the implications for aging and cancer. PMID- 12890858 TI - Salivary gland involvement in rheumatoid arthritis and its relationship to induced oxidative stress. AB - OBJECTIVES: The objective of the present study was to analyse salivary gland and free radical involvement in rheumatoid arthritis (RA). METHODS: Thirty-four consenting RA patients (rheumatoid factor-positive) and 18 healthy controls, matched in age and gender, participated in the study. Plasma and saliva samples were harvested and subjected to compositional analysis and various free radical related tests. RESULTS: The mean salivary flow rate was lower in the RA patients than in the control group, whereas all plasma and salivary antioxidants were increased. Mean values of plasma malondialdehyde and ceruloplasmin were higher in the RA patients. CONCLUSIONS: The effects of RA on salivary gland flow rates and antioxidant compositional parameters may be of great importance for the further elucidation of the role of free radicals in RA pathogenesis and for its general diagnosis and evaluation. The demonstrated correlation between the altered salivary parameters and the severity of the disease may indicate that evaluation of the salivary status of RA patients is warranted. PMID- 12890859 TI - Acupuncture for chronic low back pain in older patients: a randomized, controlled trial. AB - OBJECTIVE: To determine if acupuncture is an effective, safe adjunctive treatment to standard therapy for chronic low back pain (LBP) in older patients. METHODS: The inclusion criteria for subjects were: (i) LBP > or =12 weeks and (ii) age > or =60 yr; the exclusion criteria were (i) spinal tumour, infection or fracture and (ii) associated neurological symptoms. The subjects were randomized to two groups. The control group of subjects continued their usual care as directed by their physicians, i.e. NSAIDs, muscle relaxants, paracetamol and back exercises. Subjects in the acupuncture group in addition received biweekly acupuncture with electrical stimulation for 5 weeks. Outcome was measured by the modified Roland Disability Questionnaire (RDQ) at weeks 0, 2, 6 and 9. The primary outcome measure was change in RDQ score between weeks 0 and 6. RESULTS: Fifty-five patients were enrolled, with eight drop-outs. Twenty-four subjects were randomized to the acupuncture group and 23 were randomized to the control group. Acupuncture subjects had a significant decrease in RDQ score of 4.1 +/- 3.9 at week 6, compared with a mean decrease of 0.7 +/- 2.8 in the control group (P = 0.001). This effect was maintained for up to 4 weeks after treatment at week 9, with a decrease in RDQ of 3.5 +/- 4.4 from baseline, compared with 0.43 +/- 2.7 in the control group (P = 0.007). The mean global transition score was higher in the acupuncture group, 3.7 +/- 1.2, indicating greater improvement, compared with the score in the control group, 2.5 +/- 0.9 (P < 0.001). Fewer acupuncture subjects had medication-related side-effects compared with the control group. CONCLUSIONS: Acupuncture is an effective, safe adjunctive treatment for chronic LBP in older patients. PMID- 12890860 TI - Interleukin 1alpha, interleukin 1beta and interleukin 1 receptor gene polymorphisms in psoriatic arthritis. AB - OBJECTIVES: To investigate polymorphisms of interleukin (IL) 1alpha, IL-1beta and IL-1 receptor R1 genes in patients with psoriatic arthritis (PsA), their relationship to the age of onset of psoriasis and the pattern of joint involvement. METHODS: One hundred and forty well-characterized patients with PsA were studied. One hundred healthy controls were recruited from primary care. All were genotyped for single-nucleotide polymorphisms in the genes for IL-1alpha (position -889), IL-1beta (position +3953) and IL-1R1 (position +970). The frequencies of the respective variants were compared between patients and controls and in relation to age of onset of psoriasis, to clinical subsets of the disease and to the presence of erosions. RESULTS: All three polymorphisms were in Hardy-Weinberg equilibrium in both patients and controls. The frequency of IL 1alpha -889 CC homozygotes was significantly increased in PsA patients compared with normal controls [58 vs 40%, odds ratio (OR) 2.06, 95%, confidence interval (CI) 1.22-3.47]. The frequency of the IL-1alpha -889 C allele was significantly increased in PsA patients compared with controls (75 vs 65%, OR 1.65, 95% CI 1.11 2.45). In subset analysis there were no other significant differences in allelic frequencies for the IL-1alpha -889 C/T, IL-1beta +3953 C/T and IL-1R1 +970 C/T polymorphisms. CONCLUSIONS: The IL-1 gene complex may play a role in the development of PsA and/or psoriasis or act as a marker for other genes on chromosome 2q12 to 2q13. PMID- 12890861 TI - Modelling the cost-effectiveness of etanercept in adults with rheumatoid arthritis in the UK. AB - OBJECTIVES: This model examines the cost-effectiveness of etanercept monotherapy under British Society for Rheumatology guidelines, i.e. adults previously failing two disease-modifying anti-rheumatic drugs (DMARDs). It compares a DMARD sequence with etanercept third line against the same sequence excluding etanercept. METHOD: The 6-monthly trend in Health Assessment Questionnaire (HAQ) disability score is simulated for 10 000 patients' lifetimes using clinical trial data and published literature. Switching to the next treatment is triggered by lack of response, loss of efficacy or adverse events. Patient mortality depends on rheumatoid arthritis life-tables and on epidemiological evidence relating reduced risk to HAQ improvement. Regression of HAQ/EuroQol (EQ-5D) utility provides quality-adjusted life years (QALY) gained. Primary analysis includes drug costs, monitoring and hospitalizations. RESULTS: The central estimate cost per QALY is pound 16 330. Sensitivity analyses ( pound 7800 to pound 42 000) showed long-term HAQ progression (etanercept, DMARDs, non-responders) as most sensitive variables. The inclusion of potential avoided nursing home admissions and indirect costs/lost employment further improves the cost-effectiveness. CONCLUSIONS: For adults in the UK, the results suggest that etanercept is cost-effective when compared with non-biologic agents. The National Institute for Clinical Excellence has accepted that etanercept is cost-effective and recommended its availability for use in patients who have failed at least two DMARDs. This model was an important component of that decision. The model is further suitable for use for a wide range of other cost-effectiveness questions in rheumatoid arthritis. PMID- 12890862 TI - The Behcet's disease activity index. AB - OBJECTIVE: To identify a subset of clinical features of Behcet's disease (BD) that can be summated to form an overall index of disease activity appropriate for clinical and research use internationally. METHODS: Completed Behcet's Disease Current Activity Forms were collected from a total of 524 patients with BD from five countries. The data from 14 questions on the form were subjected to Rasch analysis to establish whether these items form a hierarchical and unidimensional scale of disease activity, both within and between countries. RESULTS: The data showed a good fit to the Rasch model within three countries using a dichotomous scoring function. However, when the data from these three countries were pooled, the fit to the model was poor. Cross-cultural differential item functioning (DIF) was found in seven items in the pooled data. When the items with DIF by country were separated and two items were removed, the resulting 26-item scale showed a good fit to the Rasch model. CONCLUSIONS: Within Turkey, Korea and the UK, the 14 items can be summated to give an index of disease activity. Analysis of the pooled data confirmed that the index is not suitable for comparison between countries or for pooling of data in the raw form, but after fitting the data to the Rasch model such comparisons can be made. This gives a scaling tool that is quick and easy to use in the clinical situation. PMID- 12890863 TI - The effect of transforming growth factor beta1 gene polymorphisms in ankylosing spondylitis. AB - OBJECTIVES: To determine whether genetic polymorphisms in or near the transforming growth factor beta1 (TGFB1) locus were associated with susceptibility to or severity of ankylosing spondylitis (AS). METHODS: Five intragenic single-nucleotide polymorphisms (SNP) and three microsatellite markers flanking the TGFB1 locus were genotyped. Seven hundred and sixty-two individuals from 184 multiplex families were genotyped for the microsatellite markers and two of the promoter SNPs. One thousand and two individuals from 212 English and 170 Finnish families with AS were genotyped for all five intragenic SNPs. A structured questionnaire was used to assess the age of symptom onset, disease duration and disease severity scores, including the BASDAI (Bath Ankylosing Spondylitis Disease Activity Index) and BASFI (Bath Ankylosing Spondylitis Functional Index). RESULTS: A weak association was noted between the rare TGFB1 +1632 T allele and AS in the Finnish population (P = 0.04) and in the combined data set (P = 0.03). No association was noted between any other SNPs or SNP haplotype and AS, even among those families with positive non-parametric linkage scores. The TGFB1 +1632 polymorphism was also associated with a younger age of symptom onset (English population, allele 2 associated with age of onset greater by 4.2 yr, P = 0.05; combined data set, allele 2 associated with age of onset greater by 3.2 yr, P = 0.02). A haplotype of coding region SNPs (TGFB1 +869/+915+1632 alleles 2/1/2) was associated with age of symptom onset in both the English parent-case trios and the combined data set (English data set, haplotype 2/1/2 associated with age of onset greater by 4.9 yr, P = 0.03; combined data set, haplotype 2/1/2 associated with greater age of onset by 4.2 yr, P = 0.006). Weak linkage with AS susceptibility was noted and the peak LOD score was 1.3 at distance 2 cM centromeric to the TGFB1 gene. No other linkage or association was found between quantitative traits and the markers. CONCLUSION: This study suggests that the polymorphisms within the TGFB1 gene play at most a small role in AS and that other genes encoded on chromosome 19 are involved in susceptibility to the disease. PMID- 12890864 TI - Characterization of myotubularin-related protein 7 and its binding partner, myotubularin-related protein 9. AB - Myotubularin-related protein 7 (MTMR7) is a member of the myotubularin (MTM) family. The cDNA encoding the mouse MTMR7 contains 1,983 bp, and the predicted protein has a deduced molecular mass of 75.6 kDa. Northern and Western blot analyses showed that MTMR7 is expressed mainly in brain and mouse neuroblastoma N1E-115 cells. Recombinant MTMR7 dephosphorylated the D-3 position of phosphatidylinositol 3-phosphate and inositol 1,3-bisphosphate [Ins(1,3)P2]. The substrate specificity of MTMR7 is different than other MTM proteins in that this enzyme prefers the water-soluble substrate. Immunofluorescence showed that MTMR7 is localized in Golgi-like granules and cytosol, and subcellular fractionation showed both cytoplasmic and membrane localization of MTMR7 in N1E-115 cells. An MTMR7-binding protein was found in an anti-MTMR7 immunoprecipitate from N1E-115 cells and identified as MTM-related protein 9 (MTMR9) by tandem mass spectrometry. The coiled-coil domain of MTMR9 was sufficient for binding to MTMR7. The binding of MTMR9 increased the Ins(1,3)P2 phosphatase activity of MTMR7. Our results show that MTMR7 forms a complex with MTMR9 and dephosphorylates phosphatidylinositol 3-phosphate and Ins(1,3)P2 in neuronal cells. PMID- 12890865 TI - Modeling the optimum duration of antibiotic prophylaxis in an anthrax outbreak. AB - A critical consideration in effective and measured public health responses to an outbreak of inhalational anthrax is the optimum duration of antibiotic prophylaxis. We develop a competing-risks model to address the duration of antibiotic prophylaxis and the incubation period that accounts for the risks of spore germination and spore clearance. The model predicts the incubation period distribution, which is confirmed by empirical data. The optimum duration of antibiotic prophylaxis depends critically on the dose of inhaled spores. At high doses, we show that exposed persons would need to remain on antibiotic prophylaxis for at least 4 months, and considerable morbidity would likely occur before antibiotic prophylaxis could even be initiated. At very low doses, 60 days of antibiotic prophylaxis is adequate. Exposure doses can be estimated from the cumulative attack rate up to the point antibiotic prophylaxis begins. The model explains that whereas < or =60 days of antibiotics were enough to protect persons in the 2001 U.S. outbreak, because doses were very low, at moderate or high doses considerably longer durations would be necessary to adequately protect exposed populations. PMID- 12890866 TI - Manganese activation of superoxide dismutase 2 in Saccharomyces cerevisiae requires MTM1, a member of the mitochondrial carrier family. AB - Manganese-containing superoxide dismutase (SOD2) plays a critical role in guarding against mitochondrial oxidative stress and is essential for survival of many organisms. Despite the recognized importance of SOD2, nothing is known regarding the mechanisms by which this nuclear-encoded protein is converted to an active enzyme in the mitochondrial matrix. To search for factors that participate in the posttranslational activation of SOD2, we screened for yeast genes that when mutated lead to SOD2 inactivation and identified a single ORF, YGR257c. The encoded protein localizes to the mitochondria and represents a member of the yeast mitochondrial carrier family. YGR257c was previously recognized as the homologue to human CGI-69, a widely expressed mitochondrial carrier family of unknown function. Our studies suggest a connection with SOD2, and we have named the yeast gene MTM1 for manganese trafficking factor for mitochondrial SOD2. Inactivation of yeast MTM1 leads to loss of SOD2 activity that is restored only when cells are treated with high supplements of manganese, but not other heavy metals, indicative of manganese deficiency in the SOD2 polypeptide. Surprisingly, the mitochondrial organelle of mtm1 Delta mutants shows no deficiency in manganese levels. Moreover, mtm1 Delta mutations do not impair activity of a cytosolic version of manganese SOD. We propose that Mtm1p functions in the mitochondrial activation of SOD2 by specifically facilitating insertion of the essential manganese cofactor. PMID- 12890867 TI - Expression of BCR/ABL and BCL-2 in myeloid progenitors leads to myeloid leukemias. AB - Chronic myelogenous leukemia is a myeloproliferative disorder (MPD) that, over time, progresses to acute leukemia. Both processes are closely associated with the t(9;22) chromosomal translocation that creates the BCR/ABL fusion gene in hematopoietic stem cells (HSCs) and their progeny. Chronic myelogenous leukemia is therefore classified as an HSC disorder in which a clone of multipotent HSCs is likely to be malignantly transformed, although direct evidence for malignant t(9;22)+ HSCs is lacking. To test whether HSC malignancy is required, we generated hMRP8p210BCR/ABL transgenic mice in which expression of BCR/ABL is absent in HSCs and targeted exclusively to myeloid progenitors and their myelomonocytic progeny. Four of 13 BCR/ABL transgenic founders developed a chronic MPD, but only one progressed to blast crisis. To address whether additional oncogenic events are required for progression to acute disease, we crossed hMRP8p210BCR/ABL mice to apoptosis-resistant hMRP8BCL-2 mice. Of 18 double-transgenic animals, 9 developed acute myeloid leukemias that were transplantable to wild-type recipients. Taken together, these data indicate that a MPD can arise in mice without expression of BCR/ABL in HSCs and that additional mutations inhibiting programmed cell death may be critical in the transition of this disease to blast-crisis leukemia. PMID- 12890868 TI - The impact of cutting balloon angioplasty for the treatment of diffuse in-stent restenosis. AB - OBJECTIVE: To determine the comparable safety and efficacy of cutting balloon angioplasty (CBA) and balloon angioplasty (BA) for diffuse in-stent restenosis (ISR). BACKGROUND: Previous studies have found that diffuse-type ISR predicts recurrence of ISR. The efficacy of CBA in this clinical setting is incompletely documented. METHODS AND RESULTS: One-hundred and seven consecutive diffuse ISR lesions (length > 10 mm) in 104 patients treated by CBA or BA were retrospectively analyzed. CBA was used in 61 lesions and BA in 46 lesions. Initial success (residual restenosis 30% with no major complications) was achieved in 99% of cases in each group. Follow-up angiography was performed in all patients at 131 93 days. Although the percent diameter stenosis after CBA was lower than after BA (19 12% versus 27 12%; p = 0.001), intravascular ultrasound (IVUS) or other QCA data were similar in the 2 groups. In follow-up angiography, recurrent restenosis rate was not statistically different between CBA (34%) and BA (52%); however, recurrent diffuse ISR was lower in the CBA group (20%) than the BA group (48%; p = 0.01). Lesion length at follow-up in the CBA group was smaller compared to the BA group (15.9 7.3 mm versus 24.2 14.4 mm; p < 0.05), while lesion length before procedure was similar. CONCLUSION: CBA for ISR is safe and effective with favorable long-term outcomes. This effect is mainly due to shortening lesion length. PMID- 12890869 TI - Cutting or not cutting: more than 10 years later the cutting balloon is around and still looking for the indication! PMID- 12890870 TI - Comparison of three-year clinical outcome of the multi-link stent with the Palmaz Schatz stent in Japanese patients with coronary artery disease: a case control study. AB - Results of trials using the ACS Multi-Link (ML) stent, one of the new generation stents, were similar to or slightly better than those of trials using the Palmaz Schatz (PS) stent. The purpose of this study was to compare long-term (3-year) clinical outcomes of patients with coronary artery disease treated with the ML stent to those treated with the PS stent. The present study consisted of 52 patients who underwent successful coronary ML stent implantation (ML group) and 52 matched control patients who underwent successful coronary PS stent implantation (PS group) from October 1997 to September 1999. During follow-up periods, cardiac events occurred in 11 patients (21%) in the ML group and 14 patients (27%) in the PS group, respectively (p = NS). Angiographic restenosis rates of American College of Cardiology/American Heart Association (ACC/AHA) lesion type A or B1 were 8.3% in both groups, and ACC/AHA lesion type B2 or C were 39.3% in the ML group and 35.7% in the PS group, respectively (p = NS). In addition, angiographic restenosis rates of ACC/AHA lesion type A or B1 were significantly lower than those of lesion type B2 or C in both groups. The results of the present study suggest that 6-month angiographic and 3-year clinical outcomes in patients with coronary artery disease treated by coronary stenting with the ML stent were comparable to those with the PS stent. PMID- 12890871 TI - Balloon sizing of pulmonary branch stenosis: a useful method to guide stent implantation. AB - Among patients with congenital heart disease, pulmonary branch stenosis is a common indication for stent implantation. Selective calibrated angiography is the standard method of vessel sizing to guide angioplasty balloon and stent selection. Our aim was to compare vessel dimensions from standard calibrated selective angiography with those obtained using a compliant sizing balloon catheter. METHODS: 9 patients with 11 pulmonary branch stenoses underwent selective calibrated angiography. Amplatzer sizing balloon catheters positioned across the stenoses were inflated with dilute contrast agent. Digital angiograms were repeated in the same projections. Measurements from both methods were analyzed statistically. Minimum, maximum proximal and maximum distal vessel diameters were all significantly larger (p < 0.01) when measured by the sizing balloon method. Angioplasty balloons and stent diameters were chosen according to the sizing balloon measurements. In 7 of 8 stented lesions, larger angioplasty balloon diameters were selected for stent implantation than would have been chosen by standard angiography. Calibrated selective angiography may undersize vessel diameters. Use of a compliant sizing balloon appears to offer an accurate method to guide stent implantation in pulmonary branch stenosis. PMID- 12890872 TI - Coronary intervention with a heparin-coated stent and aspirin only. AB - OBJECTIVES: To determine the safety and efficacy of a post-stenting anti-platelet regimen of aspirin without additional ticlopidine or clopidogrel after successful heparin-coated stent implantation. METHODS: A prospective, non randomized, multi-center pilot study of patients undergoing percutaneous coronary intervention, including those with acute coronary syndromes and small vessels with one-month clinical follow-up, was undertaken. Patients received a heparin coated stent and were treated with aspirin only. RESULTS: Over a period of 6 months, a total of 122 patients were recruited in 6 centers. Their mean age was 57.2 10.0 years, 79% were male and 31% had unstable angina. Most (75%) had single-vessel disease, predominantly of the left anterior descending artery (51%), with a mean reference diameter of 2.44 mm 0.44 mm at baseline and 2.48 0.41 mm post stenting. At a 1-month clinical follow-up, no major adverse cardiovascular events (including subacute stent thrombosis) had occurred. Five patients were readmitted to hospital for symptoms unrelated to the interventional procedure. CONCLUSIONS: Heparin-coated stent implantation using an antiplatelet regimen of aspirin only, appears to be safe and feasible. A randomized trial of a larger number of patients appears warranted. PMID- 12890873 TI - Preventing subacute stent thrombosis--is there a role for heparin-coated stents? PMID- 12890874 TI - Defining the healthcare market--a critical precursor to your strategic plan. PMID- 12890875 TI - Successful percutaneous coronary angioplasty via a solitary coronary ostium in the aorta (single coronary artery). PMID- 12890876 TI - Distal embolization during directional coronary atherectomy. AB - We report 2 cases of directional coronary atherectomy performed by using the Percu-Surge System to prevent distal embolization. A discrete amount of material was aspirated after the procedure. No clinical sequelae occurred in the immediate follow-up. These cases underline the problem of distal embolization during directional atherectomy, and the possibility to prevent it by using protection devices. PMID- 12890877 TI - Iatrogenic deep musculocutaneous radiation injury following percutaneous coronary intervention. AB - Radiation-induced skin injury has been reported for multiple fluoroscopic procedures. Previous studies have indicated that prolonged fluoroscopic exposure during even a single percutaneous coronary intervention (PCI) may lead to cutaneous radiation injury. We document a novel case of deep muscle damage requiring wide local debridement and muscle flap reconstruction in a 59-year-old man with a large radiation-induced wound to the lower thoracic region following 1 prolonged PCI procedure. The deep muscular iatrogenic injury described in this report may be the source of significant morbidity. Recommendations to reduce radiation-induced damage include careful examination of the skin site before each procedure, minimized fluoroscopy time, utilization of pulse fluoroscopy, employment of radiation filters, and collimator s and rotation of the location of the image intensifier. PMID- 12890878 TI - ICUS-guided obliteration of a complex coronary artery aneurysm with a PTFE covered stent. PMID- 12890879 TI - Complex transradial three vessel brachytherapy in a single session. AB - BACKGROUND: We report the case of a patient who underwent transradial brachytherapy in 3 different coronary vessels during a single session. She initially presented with unstable angina 4 months after the index procedure; control angiography showed severe and diffuse in-stent restenosis in the LAD, Cx and Mg arteries. METHODS: After successful dilatation of the three vessels, we performed vascular brachytherapy using the Novoste Beta-Rail system and a 60 mm length source train of 90Sr/Y radioactive seeds. No further stent was implanted. The patient left the hospital the next day. Follow-up angiography revealed widely patent vessels with no restenosis. CONCLUSION: Transradial multivessel brachytherapy can be done during the same session. PMID- 12890880 TI - Low-molecular-weight heparins and glycoprotein IIb/IIIa inhibitors with percutaneous coronary intervention in acute coronary syndromes. PMID- 12890881 TI - Treatment of bifurcation in-stent restenotic lesions with beta radiation using strontium 90 and sequential positioning pullback technique: procedural details and clinical outcomes. AB - BACKGROUND: In-stent restenotic lesions have been problematic for many patients with the need for multiple repeat percutaneous coronary interventions (PCI). The need for repeat PCI has been significantly reduced in patients since the advent of vascular brachytherapy. In-stent restenosis resulting in bifurcation presents even more of a challenge. The use of radiation therapy for the treatment of this kind of lesion has not yet been reported. The purpose of this paper is to present five cases of radiation therapy in bifurcation in-stent restenotic lesions using the intraluminal beta radiation catheter delivery system (Beta-Cath System, Novoste Corporation, Norcross, Georgia). METHODS: We reviewed the database of patients enrolled in our Compassionate Use Registry between August 1999 and April 2002. The data is reported for 5 patients who received radiation in both branches of bifurcation lesions with the Beta-Cath catheter system. RESULTS: The mean diameter of the vessels was 3.1 mm 0.5 mm. The dose administered was from 18.3 to 23 Gy, with an overlap of 3.3 to 10.3 mm; the hinge angle between the branches went from 43.3 to 65.4 . Angiographic follow-up was obtained at 6 months in 4 patients, with a single patient showing a focal (< 5 mm) edge lesion treated by balloon angioplasty (TVR no TLR). No aneurysms or zones of ectasia were noted. CONCLUSION: Beta radiation with the Beta-Cath catheter system appears to be safe, secure and clinically useful in in-stent restenotic bifurcation lesions. PMID- 12890882 TI - Macroprolactinemia: a new cause of hyperprolactinemia. AB - Hyperprolactinemia is not only seen in pregnancy but also in several pathological conditions such as prolactin (PRL) secreting pituitary adenoma (prolactinoma), intracranial tumors compressing the pituitary stalk or hypothalamus, and PRL stimulative drugs. However, some patients with hyperprolactinemia are diagnosed as having idiopathic hyperprolactinemia because the causes are unknown. They are subjected to repeated radiological examinations to find a microadenoma, to a long term treatment with bromocriptine, and even to a surgical intervention. There is accumulating evidence that macroprolactinemia, in which most circulating PRL forms large protein complexes (more than 150 kDa), is a major cause of idiopathic hyperprolactinemia. The patients with macroprolactinemia are clinically characterized by the lack of hyperprolactinemia-related symptoms such as amenorrhea and galactorrhea. We found that anti-PRL autoantibody is a leading cause of macroprolactinemia that might be heterogeneous in nature. Most patients with anti-PRL autoantibodies were symptom-free and pregnancy was possible despite a marked hyperprolactinemia. Identification of macroprolactinemia is clinically important to prevent unnecessary examinations and treatments in patients with idiopathic hyperprolactinemia. PMID- 12890883 TI - Current research on methamphetamine-induced neurotoxicity: animal models of monoamine disruption. AB - Methamphetamine (METH)-induced neurotoxicity is characterized by a long-lasting depletion of striatal dopamine (DA) and serotonin as well as damage to striatal dopaminergic and serotonergic nerve terminals. Several hypotheses regarding the mechanism underlying METH-induced neurotoxicity have been proposed. In particular, it is thought that endogenous DA in the striatum may play an important role in mediating METH-induced neuronal damage. This hypothesis is based on the observation of free radical formation and oxidative stress produced by auto-oxidation of DA consequent to its displacement from synaptic vesicles to cytoplasm. In addition, METH-induced neurotoxicity may be linked to the glutamate and nitric oxide systems within the striatum. Moreover, using knockout mice lacking the DA transporter, the vesicular monoamine transporter 2, c-fos, or nitric oxide synthetase, it was determined that these factors may be connected in some way to METH-induced neurotoxicity. Finally a role for apoptosis in METH induced neurotoxicity has also been established including evidence of protection of bcl-2, expression of p53 protein, and terminal deoxynucleotidyl transferase mediated dUTP nick-end labeling (TUNEL), activity of caspase-3. The neuronal damage induced by METH may reflect neurological disorders such as autism and Parkinson's disease. PMID- 12890884 TI - Pharmacological evidence for involvement of phospholipase D, protein kinase C, and sodium-calcium exchanger in alpha-adrenoceptor-mediated negative inotropy in adult mouse ventricle. AB - The intracellular signalling pathway for alpha-adrenoceptor-mediated negative inotropy was studied pharmacologically in isolated adult mouse ventricle. The negative inotropy was inhibited by GF-109203X, a nonselective protein kinase C inhibitor. Phorbol 12-myristate 13-acetate also produced sustained negative inotropy, which was inhibited by KB-R7943, a Na(+)/Ca(2+) exchanger inhibitor. The alpha-adrenoceptor-mediated negative inotropy was augmented by RHC-80267, a diacylglycerol lipase inhibitor, but was inhibited either by C(2)-ceramide, a phospholipase D inhibitor, and high concentration of propranolol (50 micro M), which inhibits phosphatidate phosphohydrolase. The inotropy was not affected by U 73122, a phospholipase C inhibitor. Lavendustin-A, a tyrosine kinase inhibitor, also inhibited the negative inotropy. These findings suggest that alpha adrenoceptor-mediated negative inotropy in adult mouse ventricle is mediated by activation of tyrosine kinase, the phospholipase D-phosphatidate phosphohydrolase pathway, and protein kinase C. PMID- 12890885 TI - The epoxy group of pramanicin is required for the optimal endothelium-dependent relaxation of rat aorta. AB - The vascular effects of a newly discovered anti-fungal agent, pramanicin (PMC), and its two analogues, PMC-A, in which the epoxy group is replaced by a - HC = CH - bond, and PMC-B, on which the diene is converted to the saturated (CH(2))(4) derivative, respectively, were investigated in rat aorta. All three compounds caused an initial endothelial-dependent relaxation, which is prevented either by removal of endothelium or inclusion of the nitric oxide synthase inhibitor L NAME. Upon prolonged incubation with the aortic rings, they also caused endothelial cell dysfunction characterized as reduced relaxation to carbachol (CCh). These effects were the strongest for PMC, being completely inhibitory at 20 microM after 30 min incubation, whereas those of PMC-A and PMC-B were smaller and comparable with each other, causing 30 - 40% inhibition at 20 micro M. PMC and its analogues had no effect on KCl-induced contraction and also had no effect on relaxation induced by sodium nitroprusside, suggesting that these compounds had no effect on the basic mechanisms of the contractile elements. Phenylephrine (PE)-induced contraction, however, was significantly reduced in the presence of these compounds, the inhibitory effect being strongest with PMC, but this inhibitory action was rapidly reversible and not of the competitive mode with respect to PE. We conclude that the epoxy group in PMC is required for the optimal vascular effects. We have discussed and speculated upon the possible mechanisms of action of PMC. The potent, selective, and irreversible inhibitory effect of PMC on the endothelial function points to its potential development into an anti-angiogenic drug. PMID- 12890886 TI - Effects of muscarinic receptor antagonists with or without M2 antagonist activity on cholinergic reflex bronchoconstriction in ovalbumin-sensitized and -challenged mice. AB - To investigate whether the inhibition of muscarinic M(2) receptors results in the enhancement of reflex bronchoconstriction under airway hyperresponsiveness, we evaluated the effects of muscarinic antagonists with or without M(2) antagonist activity on methacholine (MCh)- and SO(2)-induced airway responses in ovalbumin (OVA)-sensitized and -challenged mice. In this model, similar airway hyperresponsiveness to MCh (12 mg/ml) was observed on Days 31 and 37 (2.2-fold and 2.7-fold, respectively). However, airway hyperresponsiveness to SO(2) (0.05 l/min) on Day 37 was less than that on Day 31 (4.0- and 2.7-fold on Days 31 and 37), indicating reflex bronchoconstriction was enhanced on Day 31 in comparison to Day 37. Ipratropium (0.03 - 0.3 mg/ml, inhalation) and Compound A (0.1 - 3 mg/kg, p.o.) inhibited MCh-induced responses on Days 31 and 37. Although ipratropium (0.03 - 1 mg/ml) dose-dependently inhibited SO(2)-induced responses on Day 31, ipratropium at a dose of 0.1 mg/ml significantly increased SO(2) induced responses on Day 37 (162.2% of the corresponding control). On the other hand, Compound A (0.03 - 0.3 mg/kg, p.o.) inhibited SO(2)-induced responses without any increases on Days 31 and 37. These results suggest that two different conditions of reflex bronchoconstriction are presented in this model: 1) SO(2) induced responses are enhanced by dysfunctional M(2) receptors on Day 31; 2) the dysfunctional M(2) receptors are partially restored on Day 37. In addition, the inhibition of the restored M(2) receptors further enhance reflex bronchoconstriction. PMID- 12890887 TI - Pharmacological characterization of vasorelaxant effects of BMS-180448, a novel cardioselective ATP-sensitive potassium channel opener, in rat aorta. AB - This study was designed to characterize vasorelaxant effects of BMS-180448 ((3S trans)-N-(4-chlorophenyl)-N'-cyano-N"-(6-cyano-3,4-dihydro-3-hydroxy-2,2-dimethyl 2H-1-benzopyran-4-yl)), a prototype cardioselective ATP-sensitive potassium channel opener, in rat aorta. BMS-180448 relaxed phenylephrine-precontracted endothelium-intact aortic rings (IC(50): 0.97 +/- 0.29 micro M), the effect being significantly attenuated by removal of functional endothelium (IC(50): 1.95 +/- 0.23 micro M) and pretreatment with N(G)-nitro-L-arginine methyl ester (L-NAME) or methylene blue. BMS-180448 completely relaxed endothelium-denuded aorta contracted with phorbol 12,13-dibutyrate, PGF(2)(alpha), and U46619 with a significantly greater potency (IC(50): 0.069 +/- 0.002, 0.055 +/- 0.002, and 0.068 +/- 0.008 micro M, respectively, P<0.05) than that contracted with phenylephrine (1.95 +/- 0.23 micro M) or KCl (0.25 +/- 0.08 micro M), indicating potency change with the type of vasoconstrictor. BMS-180448 (1 - 3 micro M) inhibited Ca(2+) (0.5 - 2.5 mM)-induced contraction of endothelium-denuded aorta evoked in the presence of high KCl (65.4 mM), but had no effect on contraction induced by phenylephrine in Ca(2+)-free buffer. BMS-180448 (10 micro M) increased cAMP level in aorta by approximately two-fold compared with the control, comparable to forskolin, an adenylate cyclase activator. These findings suggest that cardioselective BMS-180448 still exerts significant vasorelaxant activity in rat aorta contracted with various vasoconstrictors via multiple mechanisms including the blockade of extracellular Ca(2+) influx through voltage-dependent channels and activation of the adenylate cyclase and nitric oxide pathway, with the possibility of hemodynamic implications in certain clinical conditions such as myocardial infarction and hypertension. PMID- 12890888 TI - Possible involvement of both endoplasmic reticulum-and mitochondria-dependent pathways in thapsigargin-induced apoptosis in human neuroblastoma SH-SY5Y cells. AB - Recently, it has been shown that endoplasmic reticulum (ER) stress causes apoptosis. However, the mechanism of the ER stress-dependent pathway is not fully understood. In human neuroblastoma SH-SY5Y cells, we detected a caspase-12-like protein that has a molecular mass (approximately 60 kDa) similar to that of mouse caspase-12. Thapsigargin, an inhibitor of ER-associated Ca(2+)-ATPase, induced the degradation of caspase-12-like protein. In addition, the degradation of caspases-9 and -3, cleavage of poly(ADP-ribose) polymerase, DNA fragmentation, and cell death were also observed. Pretreatment with phorbol-12-myristate-13 acetate, which induces the expression of antiapoptotic Bcl-2, inhibited thapsigargin-induced degradation of caspases-9 and -3, but not caspase-12-like protein degradation. A caspase inhibitor, benzyloxycarbonyl-Val-Ala-Asp(OCH(3)) CH(2)F, inhibited the degradation of caspase-12-like protein, but not that of caspases-9 and -3. These results suggest that thapsigargin may induce the activation of both ER- and mitochondria-dependent pathways in human SH-SY5Y cells. PMID- 12890889 TI - The influence of contextual cue on antinociceptive tolerance and facilitation of memory with morphine. AB - The response latencies of the biting response in the tail-pinch test and step through response in the passive avoidance test were measured in mice. 1) Scopolamine decreased the latency of the step-through response in the passive avoidance test, but diazepam did not. 2) Morphine (5 mg/kg, i.p.) was given 30 min before the test for 4 consecutive days in novel environments by sessions. The decrease in biting response latency was attenuated and delayed in the groups in which contexts were changed daily compared to the groups that were maintained in the same context throughout the conditioning period. The latency of step-through response was increased with morphine in both groups regardless of the change or lack of change in context. 3) Morphine was repeatedly administered to animals in the same environment and then the context was changed. The decreased latency in the tail-pinch test was significantly reversed by the change in context, but the response in the passive avoidance test maintained a longer latency. These findings indicate that morphine develops associative and nonassociative antinociceptive tolerance, indicating that antinociceptive tolerance to morphine has contextual specificity, but the facilitation of memory does not. The results indicate that the facilitation of memory with morphine may contribute to associative learning in antinociceptive tolerance to morphine. PMID- 12890890 TI - Expression of group I metabotropic glutamate receptors in rat hippocampal cells in culture and their characterization by intracellular calcium ion dynamics. AB - The distribution of group I metabotropic glutamate receptors in rat hippocampal cells in culture was examined by calcium imaging and immunocytochemistry. To distinguish different cell types in the culture, the effects of t-ACPD ((1S,3R)-1 aminocyclopentane-1,3-dicarboxylic acid) and of NMDA (N-methyl-D-aspartate) were examined. About 40% of the cultured cells showed either a transient increase or a sustained or oscillatory increase in the intracellular calcium concentration ([Ca(2+)](i)) during t-ACPD administration, while about 60% of the cells showed a sustained [Ca(2+)](i) increase in response to NMDA. Cells that showed an oscillatory [Ca(2+)](i) change during t-ACPD administration did not respond to NMDA administration, while cells that showed a sustained [Ca(2+)](i) increase during NMDA administration did not show any oscillatory response to t-ACPD. After pharmacological examination using those two agonists, the cultured cells were subjected to immunocytochemistry using anti-GFAP and ant-MAP-2 antibodies to distinguish, respectively, astrocytes and neurons. All cells responding to NMDA with a sustained [Ca(2+)](i) increase were MAP-2-positive, whereas all cells showing either oscillatory or sustained [Ca(2+)](i) increase in response to t ACPD were GFAP-positive. The present results show that, in these cultures, group I metabotropic glutamate receptors are mainly expressed on glial cells and contribute to dynamic [Ca(2+)](i) changes in astrocytes. PMID- 12890891 TI - A novel action of palmitoyl-L-carnitine in human vascular endothelial cells. AB - Palmitoyl-L-carnitine (palcar), which accumulates in ischemic heart, affects cellular functions of vascular endothelium in the ischemic area. The aim of this study was to examine the effects of palcar on intracellular Ca(2+) concentration ([Ca(2+)](i)) in vascular endothelial cells in comparison with those of sphingosine-1-phosphate (S1P) and to investigate the underlying mechanisms. Application of palcar at a concentration range between 0.3 and 3 micro M elevated [Ca(2+)](i) in huvecs, and its potency was about 30 times lower than that of S1P. When human umbilical vein endothelial cells (huvecs) were treated with 100 ng/ml pertussis toxin (PTX) for 15 h, they failed to respond to palcar or S1P, but did respond to 3 micro M histamine (His), suggesting that the response induced by palcar as well as S1P is mediated by a PTX-sensitive GTP binding protein, Gi. Although the sensitivity to palcar and S1P varied widely among huvecs from individuals, response to 3 micro M palcar in each huvec clearly paralleled that to 0.3 micro M S1P (r = 0.79, P<0.001). On the other hand, pre-treatment of huvecs with palcar abolished subsequent S1P-induced elevation of [Ca(2+)](i), but not the His-induced elevation. Our data indicate that palcar has a novel action on huvecs as a potential agonist of receptors for S1P. Effective inhibition of the response to S1P by palcar suggests that palcar affects functions regulated by S1P. PMID- 12890892 TI - Orexin receptor type-1 couples exclusively to pertussis toxin-insensitive G proteins, while orexin receptor type-2 couples to both pertussis toxin-sensitive and -insensitive G-proteins. AB - Signal transduction pathways of orexin receptors were examined using a nerve-like cell line transfected with orexin receptor type-1 (OX1R) and orexin receptor type 2 (OX2R). Forskolin-stimulated cyclic adenosine 3,5-monophosphate (cAMP) accumulation in OX2R-expressing cells was inhibited by orexin in a dose-dependent manner, and the effect was abolished by pretreatment with pertussis toxin (PTX). The inhibitory effect of orexin on forskolin-stimulated cAMP accumulation was not observed in OX1R-expressing cells. Administration of orexin to these cells resulted in a transient increase of intracellular calcium concentration ([Ca(2+)](i)). Orexin-stimulated increases in [Ca(2+)](i) in OX1R- or OX2R expressing cells were not affected by the PTX pretreatment. These observations suggest that OX1R couples exclusively to PTX-insensitive G-proteins, while OX2R couples to both PTX-sensitive and -insensitive G-proteins. To examine the relative contributions of these G-proteins in OX2R-mediated activation of neurons, we used histaminergic tuberomammillary nucleus neurons, in which OX2R is abundantly expressed. We found that a phospholipase C (PLC)-inhibitor, U73122, inhibits orexin-mediated neuronal activation, but PTX showed no effect on it. This suggests that although OX2R couples to multiple G-proteins, activation of neurons by orexins through OX2R is mediated via a PTX-insensitive, PLC dependent pathway. PMID- 12890893 TI - High-glucose enhances a thromboxane A2-induced aortic contraction mediated by an alteration of phosphatidylinositol turnover. AB - The effect of the thromboxane A(2) analogue U46619 (9,11-dideoxy-11alpha,9alpha epoxymethanoprostaglandin F(2)(alpha)) on sustained contraction in the mouse aorta was investigated. U46619 induced concentration-dependent (1 - 100 nM) increases in contraction. These contractile responses were enhanced significantly under high-glucose-physiological salt solution (HG-PSS) (2-fold greater than normal-PSS) conditions. This hyperactivation may be associated with aortic dysfunction in diabetes. However, the mechanisms remain unclear. HG-PSS enhanced U46619-induced accumulation of endogenous diacylglycerol (DG). Phospholipase C inhibitor (U73122) suppressed DG accumulation under normal conditions; however, suppression was not observed under high-glucose conditions. The HG-PSS-induced enhancement of contraction was inhibited by protein kinase C (PKC) inhibitor (calphostin C). This result indicated that accumulated DG might increase PKC activity, which then stimulates DG kinase activation as a feedback mechanism. DG kinase inhibition also suppressed HG-PSS-induced enhancement of contraction. Increased myo-inositol incorporation was detected under high-glucose conditions, indicating an acceleration of phosphatidylinositol (PI)-turnover. Moreover, rho kinase inhibitor (Y27632) suppressed U46619-induced contraction exclusively in normal-PSS. These findings indicated that HG-PSS treatment increases DG synthesis derived from incorporated glucose, PKC and DG kinase activation, and enhances the U46619-induced contraction via acceleration of PI-turnover. This series of responses may be involved in the dysfunction of aorta under high-glucose conditions occurring in association with diabetes. PMID- 12890894 TI - Selective inhibition of mammalian DNA polymerase alpha by vitamin D2 and D3. AB - As described previously (H. Togashi et al. Biochem Pharmacol. 1998;56:583-590), the irradiated products of provitamin D(2) (ergosterol) inhibit the activities of eukaryotic DNA polymerases. In this report, therefore, we investigated whether vitamin D and its related compounds inhibited the activities of DNA polymerases. As expected, vitamin D(2) and vitamin D(3) were found to be selective inhibitors of mammalian DNA polymerase alpha (pol alpha) with IC(50) values of 123 and 96 micro M, respectively. On the other hand, provitamin D(2), provitamin D(3), and the active form of vitamin D(3) such as 1alpha,25-dihydroxyvitamin D(3) could not influence any of the DNA polymerase activities. Interestingly, vitamin D(3)-3beta sulfate was a much stronger pol alpha inhibitor with an IC(50) value of 7.1 micro M. Vitamin D(2), vitamin D(3), and vitamin D(3)-3beta-sulfate could prevent the growth of NUGC-3 human gastric cancer cells with LD(50) values of 133, 77, and 44 micro M, respectively, but provitamin D(2) and provitamin D(3) could not. The cells were halted at the G1 phase in the cell cycle by these compounds. PMID- 12890895 TI - Anti-allergic substances contained in the pollen of Cryptomeria japonica possess diverse effects on the degranulation of RBL-2H3 cells. AB - Following prolonged exposure to some of the flavonoids with RBL-2H3 cells, secretion of hexosaminidase, a granule constituent, stimulated by an immunologic was enhanced. RBL-2H3 cells do not normally respond to polybasic secretagogues, but as reported here, they do so after prolonged exposure. Effect of flavonoids on secretion of hexosaminidase was also investigated. Of the thirteen flavonoids, quercetin and fisetin were the most potent inhibitors. A structure-activity study indicated that the position, number, and substitution of the hydroxy group of the B ring and saturation of the C2-C3 bond are important factors affecting flavonoid inhibition of secretary granules in RBL-2H3 cells. PMID- 12890896 TI - Use-dependent removal of the 4-aminopyridine-induced block of the transient K+ current in rat dorsal root ganglia. AB - Effects of 4-aminopyridine (4-AP) on the transient K(+) current (I(A)) was studied in rat sensory neurons using the whole cell patch-clamp technique. The amplitude of I(A) was reduced by 4-AP. The steady-state inactivation curve for I(A) was shifted in the positive direction by 4-AP, suggesting that the blocking action of 4-AP may be attenuated by membrane depolarization. When two I(A)s were evoked with variable intervals, the peak amplitude of the I(A) induced by the second pulse was augmented in the presence of 4-AP. These results indicate that the action of 4-AP can be modulated by concurrent neuronal activities. PMID- 12890897 TI - [Role of chymase in vascular diseases and the efficacy of chymase inhibitor]. AB - In vascular tissues, angiotensin II is cleaved from angiotensin I by chymase and angiotensin converting enzyme (ACE). In the normal state, chymase is stored in mast cells and has no angiotensin II-forming activity, while chymase is activated immediately where mast cells have been activated by local stimuli. A clinical trial of an angiotensin receptor blocker (ARB) for preventing restenosis after percutaneous transluminal coronary angioplasty was successful, but that of an ACE inhibitor was not. After balloon injury in dog vessels, chymase activity was significantly increased in the injured artery, and a chymase inhibitor and an ARB were effective in preventing the vascular proliferation, but an ACE inhibitor was ineffective. In dog grafted veins, intimal area, chymase activity, and angiotensin II concentration were significantly increased after the operation, while they were significantly suppressed by a chymase inhibitor. However, the chymase inhibitor, unlike ACE inhibitor and ARB, did not affect blood pressure. These reports indicate that local angiotensin II production by chymase is involved only in the injured vessels. Therefore, a chymase inhibitor may be useful for preventing vascular disorders without affecting blood pressure. PMID- 12890898 TI - [Pharmacological study on the effects of the adenosine uptake inhibitor KF24345 on inflammatory diseases]. AB - Adenosine protects against cellular damage and dysfunction under several adverse conditions, including inflammation. We examined the effects of KF24345, a novel adenosine uptake inhibitor, on inflammatory diseases to investigate whether the adenosine uptake inhibition is useful for the treatment of inflammation. KF24345 inhibited adenosine uptake into washed erythrocytes (in vitro) and sampled blood cells from mice after its oral administration (in vivo). KF24345 significantly suppressed lipopolysaccharide-induced tumor necrosis factor-alpha production and leukopenia in mice, and the effects of KF24345 were abolished by the treatment with a non-selective or an A(2A)-selective adenosine receptor antagonist. In the experimental glomerulonephritis induced in mice by anti-glomerular basement membrane antiserum, KF24345 significantly inhibited proteinuria and glomerular damage without exhibiting the side effects observed following the treatment with prednisolone and cyclophosphamide. In addition, KF24345 ameliorated the severity of experimental acute pancreatitis induced by cerulein or choline-deficient and ethionine-supplemented diet in mice, and it decreased mortality accompanying severe acute pancreatitis. The anti-pancreatitis effects of KF24345 were abolished by the treatment with a non-selective or an A(2A)-selective adenosine receptor antagonist. These results suggest that KF24345 and adenosine uptake inhibitors can be a new therapeutic approach for various inflammatory diseases, including glomerulonephritis and acute pancreatitis. PMID- 12890899 TI - [Brain microdialysis of isolation-reared mice under freely-moving conditions]. AB - Brain microdialysis has become a well established and a widely-used technique for in vivo measurement of extracellular levels of neurotransmitters. However, a single neurotransmitter in the perfusates has been measured in most studies. The present paper describes a simultaneous measurement of serotonin and dopamine release in the prefrontal cortex of mice. Using this technique, we examined the modulation by serotonin(1A) receptors of serotonin, dopamine, and noradrenaline release in the cortex of isolation-reared mice. The isolation-reared mouse is a useful model of complicated mood disorders including phenotypes of anxiety, depression, and aggression. The study shows that isolation rearing selectively decreases the sensitivity of serotonin(1A) receptors to increase dopamine release. PMID- 12890900 TI - [Profile of the anti-tumor effects of amrubicin, a completely synthetic anthracycline]. AB - Amrubicin is a completely synthetic anthracycline derivative. In contrast, however, the anthracyclines used clinically thus far have been produced by fermentation or semisynthesis. Amrubicin is structurally distinguishable from other anthracyclines by the amino group at the 9-position and its unique sugar moiety. Amrubicinol, the C-13 hydroxy- metabolite of amrubicin, is associated with a 5 to 200 times greater cytotoxicity than amrubicin. Amrubicin exhibited superior in vivo antitumor activity to doxorubicin in the human tumor xenograft model. Using this model, the level of amrubicinol (active metabolite) was shown to be higher than that of doxorubicin in tumor tissues, but lower in normal tissues. These results suggest potent therapeutic activity for amrubicin because of the selective distribution of its highly active metabolite, amrubicinol, in tumors. These anti-tumor effects of amrubicin are considered to be induced by DNA topoisomeraseII inhibition. In clinical studies, amrubicin has demonstrated potent single agent activity as compared to a standard regimen in untreated patients with extensive small cell lung cancer. Its major toxicity was myelosuppression (especially neutropenia). PMID- 12890901 TI - [Pharmacological profile of a specific neutrophil elastase inhibitor, Sivelestat sodium hydrate]. AB - Imbalance between neutrophil elastase (NE) and its endogenous protease inhibitors has been considered to be one of possible mechanisms by which NE causes lung tissue destruction. It has been shown that the amount and/or activity of NE is increased in blood and bronchoalveolar lavage fluid in patients with acute lung injury. Accordingly, animals undergoing acute lung injury have increased NE activity such as in blood and bronchoalveolar lavage fluid. Sivelestat sodium hydrate (Sivelestat) is a synthetic inhibitor of NE with highly specificity to NE. Many studies have indicated that Sivelestat treatment improves inflammatory and edematous changes of lungs and survival as well as increased NE activity in several animal models of acute lung injury. Clinical studies have demonstrated that Sivelestat improves this injury that is associated with systemic inflammatory response syndrome. As compared with endogenous protease inhibitors that have high molecular mass, Sivelestat, a synthetic and low molecular weight elastase inhibitor, may be delivered to the inflammatory sites more easily and effectively and is considred to improve typical symptoms of acute lung injury. Clinical use of Sivelestat would further clarify the usefulness of this compound in clinical acute lung injury. PMID- 12890902 TI - [Pharmacological properties and expected clinical role of an injectable new quinolone antibiotic, pazufloxacin mesilate]. AB - Pazufloxacin mesilate (PZFX: Pasil INJECTION, Pazucross INJECTION) is a novel injectable quinolone antibiotic that was discovered by Toyama Chemical Co., Ltd. and codeveloped by Toyama Chemical Co. Ltd. and Mitsubishi Pharma Corporation. Laboratory studies indicated that PZFX reached high plasma levels quickly after intravenous administration, and it exhibited weak convulsion inducing activity, low local irritant effect, and less hypotensive activity, all of which are generally recognized as side effects associated with other injectable quinolones. PZFX has potent antibacterial and bactericidal activities against cephalosporin-, carbapenem-, and aminoglycoside-resistant strains. These favorable antibacterial and bactericidal activities gave PZFX superior therapeutic effects, compared to injectable cephalosporin antibiotics, on experimental animal infection models caused by those resistant strains. Clinical studies also revealed PZFX was as safe and effective as the injectable cephalosporin, ceftazidime, against various moderate to severe infections. Furthermore, PZFX showed good clinical effects on the infections resistant to chemotherapy by the other antimicrobial agents. These results indicate that PZFX is a viable choice for various bacterial infections. In this review, results of laboratory and clinical studies are summarized and the clinical role of PZFX among the injectable antimicrobials is discussed. PMID- 12890903 TI - Recent advances in the treatment of arrhythmias. AB - Advances in endocardial mapping techniques and ablation have greatly increased the indications for catheter-ablation of supraventricular arrhythmias. Rate or rhythm control is a valid treatment option for patients with atrial fibrillation; however, all patients with one or more risk factors should be treated with oral anticoagulants. The early success rate and long-term cure of atrial fibrillation by radiofrequency catheter ablation continues to increase. The number of centers offering this treatment option has increased substantially. Implantable defibrillator-cardioverters are the primary treatment modality for patients with ventricular tachycardia and their role in primary prevention is also being defined. Future advances in arrhythmia management will include improvements in catheter design and energy sources for ablation, and greater monitoring capacity of implantable devices. PMID- 12890904 TI - Men's Health Study: epidemiology of erectile dysfunction and cardiovascular disease. AB - The present study collected data about 6,112 Japanese male patients from 447 outpatient clinics. Of those who underwent medical examination by a general practitioner on an outpatient basis, up to 81% had some degree of erectile dysfunction (ED), ranging from mild to severe. ED was noted to be predominant among patients affected by cardiovascular disease (CVD) or diabetes mellitus (DM), and the presence of CVD increased the risk of ED. In an aging society, patients undergoing treatment for ED as part of their routine medical care are highly likely to have concomitant CVD. As shown in the present survey, clinicians need to be aware of the high incidence of ED among such patients, because ED represents a symptom originating from damage to the vascular endothelium. A total of 41% of ED patients are either willing to receive pharmacotherapy for ED or will consider treatment. Active treatment of ED with sildenafil is suitable for patients with CVD. PMID- 12890905 TI - Serum concentrations of vascular endothelial growth factor and monocyte-colony stimulating factor in peripheral arterial disease. AB - Vascular endothelial growth factor (VEGF) strongly promotes angiogenesis, and monocyte-colony stimulating factor (M-CSF) regulates the differentiation, proliferation, and survival of monocytes. Both VEGF and M-CSF are expressed in atherosclerotic lesions. The present study was performed to clarify the role of VEGF and M-CSF in the development of peripheral artery disease (PAD). The serum VEGF and M-CSF concentrations were determined in patients with arteriosclerosis obliterans (ASO) and thromboangitis obliterans (TAO). In both patient groups the serum VEGF concentrations were significantly higher than those in the control subjects. In contrast, the serum M-CSF concentrations in the ASO patients were significantly higher than those in both the TAO patients and control subjects, but there were no differences in the M-CSF concentrations between the TAO patients and control subjects. There was no correlation between the serum concentrations of VEGF and M-CSF. In conclusion, the serum VEGF concentration was increased in ASO and TAO patients, but increased concentration of M-CSF was seen only in ASO patients. These results may reflect a difference between ASO and TAO in disease pathogenesis. PMID- 12890906 TI - Efficacy of iodine-123-15-(p-iodophenyl)-3-R, S-methylpentadecanoic acid single photon emission computed tomography imaging in detecting myocardial ischemia in children with Kawasaki disease. AB - To evaluate its efficacy in detecting myocardial ischemia in children, iodine-123 labeled 15-(p-iodophenyl)-3-R, S-methylpentadecanoic acid (BMIPP) myocardial single photon emission computed tomography (SPECT) imaging was performed in 16 pediatric patients with Kawasaki disease (KD, 11 male, 5 female; mean age and range: 13 years 8 months and 8 years 11 months to 17 years 7 months). Five children with chest pain and no cardiac disease were studied as controls (2 male, 3 female; mean age and range: 13 years 4 months and 9 years 4 months to 17 years 11 months). Selective coronary angiography was also performed in the 16 patients to evaluate the location of coronary stenosis and coronary aneurysms. The SPECT images were expressed as polar maps (Bull's eye maps) and the 'defect' area was defined as where the uptake of BMIPP was less than the standardized BMIPP images of the 5 control children. In the 16 patients, 33 segments had coronary aneurysms and 10 (10/33: 30.3%) had significant coronary stenosis on selective coronary angiography. Nine of the 10 (90%) segments with significant coronary stenosis showed a defect on the BMIPP image whereas only 6 of the 23 (26.1%) segments without coronary stenosis showed a defect on BMIPP imaging. The sensitivity of BMIPP SPECT imaging for detection of coronary stenosis was 90% (9/10) and its specificity was 73.9% (17/23), whereas the sensitivity of (201)Tl SPECT imaging was 80% (8/10) and its specificity was 60% (14/23). There was no significant difference between the BMIPP and 201Tl SPECT images in either the sensitivity or specificity for the detection of coronary stenosis. In the present series, only one case had discordant BMIPP uptake (BMIPP uptake < (201)Tl uptake) in which there was a large coronary aneurysm and re-canalization after complete obstruction at segment 1 of the right coronary artery. This discordant BMIPP uptake reflects the possibility of ischemic but viable myocardium after re canalization of a large aneurysm in KD. In conclusion, BMIPP SPECT imaging is useful for detecting the areas of ischemic myocardium caused by coronary artery stenosis in children with KD. PMID- 12890907 TI - Prediction of no-reflow phenomenon after successful percutaneous coronary intervention in patients with acute myocardial infarction: intravascular ultrasound findings. AB - Plaque characterization by intravascular ultrasound (IVUS) before percutaneous coronary intervention (PCI) was evaluated in 81 consecutive patients with acute myocardial infarction (AMI) to establish if IVUS can predict the occurrence of the 'no-reflow' phenomenon. Angiographic no-reflow was defined as TIMI flow grade 1 or 2 without any mechanical obstruction in the epicardial artery. Patients were divided into 2 groups according to the post-PCI angiograms: normal flow (group R, n=60) and no-reflow (group NR, n=21). Although the incidence of either soft or noncalcified plaque was not statistically different between the groups, positive vessel remodeling was more frequent in group NR than in group R (57.1% vs 31.6%, p<0.05). Lipid core was also more frequently found in group NR than in group R (61.9% vs 25.0%, p<0.01). Positively remodeled vessels with lipid-rich plaques as characterized by IVUS before PCI predicted the occurrence of angiographic no reflow with a sensitivity of 43% and a specificity of 60%. PMID- 12890908 TI - Eccentric aortic regurgitation in patients with right coronary cusp prolapse complicating a ventricular septal defect. AB - To analyze the clinical significance of eccentric aortic regurgitation (AR) complicating the right coronary cusp prolapse associated with a ventricular septal defect (VSD), the Doppler echocardiograms of 129 patients were reviewed. In 102 patients, AR was classified as mild and in 27 patients it was classified as moderate. Eccentric AR was defined as the jet distributing in an eccentric direction. In 15 patients of the moderate group, AR was already moderate at the initial examination and of these, the AR was eccentric in 14 and central in 1. In 12 patients who initially had mild AR, it became moderate during follow-up. In 7 patients with mild, central AR, 6 worsened to central moderate AR and 1 evolved to eccentric moderate AR. Eccentric mild AR patients all developed eccentric moderate AR. Within the mild AR group, 5 of 9 patients with eccentric AR progressed from mild to moderate, whereas only 7 of 105 patients with central AR did so (p<0.01). In conclusion, eccentric AR may be an advanced finding of the AR associated with right coronary cusp prolapse in some patients, but in others eccentric AR is highly likely to progress and is malignant. PMID- 12890909 TI - Actual state of radiation exposure during coronary angioplasty: a multicenter study in the nationwide database for cost analysis of percutaneous transluminal coronary angioplasty in Japan. AB - Percutaneous coronary intervention (PCI) has been associated with excessive radiation exposure, so the present study was designed to investigate the determinants of fluoroscopic time during PCI among characteristics inherent to the patient, procedure and hospital in 388 consecutive patients between November 1996 and March 1999 in 11 hospitals included in the Nationwide Database for Cost Analysis of percutaneous transluminal coronary angioplasty in Japan. Fluoroscopic time, which reflects radiation exposure, was used as the dependent variable in a multiple linear regression analysis. The mean fluoroscopic time was 27+/-15 min. The negative value of partial correlation of the hospital code (r=-0.374) and acute myocardial infarction (-0.163) indicated that these were explanatory variables that decreased fluoroscopic time. The positive value of the number of intraaortic balloon pump catheters (r=0.144), the number of balloons (r=0.304) and hospital stay (r=0.147) indicated these were explanatory variables that increased fluoroscopic time. The absolute value of r, (r=0.374), of the hospital code was the highest. In the final R(2) (=0.304) of the model, which is the partial R-square that determined the dependent variable, the hospital code showed the highest value. In conclusion, there was an inter-hospital difference in fluoroscopic time. Although the hospital code was found to be the most powerful determinant of fluoroscopic time, the R(2) (=0.304) of the model showed so lower value that other hospital characteristics that were not included in the model may influence the fluoroscopic time. PMID- 12890910 TI - Left ventricular performance during pregnancy in patients with repaired tetralogy of Fallot: prospective evaluation using the Tei index. AB - Little is known about the influence of hemodynamic changes on ventricular performance during pregnancy in patients with congenital heart disease. A prospective evaluation was made of 17 pregnancies in 11 women with repaired tetralogy of Fallot (ToF). Twenty age-matched non-pregnant women and 41 stable pregnant women without heart disease were studied as controls. Blood pressure (BP), heart rate (HR), left ventricular end-diastolic dimension (LVEDD) and ejection fraction (LVEF), and the Tei index were measured before pregnancy; during the 1st, 2nd, and 3rd trimesters; and after delivery. BP and HR gradually increased during the pregnancy in both groups. The LVEDD in the controls increased throughout the gestation period, then decreased after delivery. In contrast, the LVEDD in the ToF was significantly smaller than that of the controls, and no such increase was observed. The LVEF did not change throughout pregnancy or after the delivery in either group. Although the Tei index of the controls did not change throughout the pregnancy, in the ToF group it was significantly increased in the 2nd and 3rd trimester and then decreased after delivery. These findings suggest that subclinical left ventricular intolerance for volume overload, probably because of the smaller LVEDD, may exist during pregnancy in patients with a repaired ToF. PMID- 12890911 TI - Specific findings of the standard 12-lead ECG in patients with 'Takotsubo' cardiomyopathy: comparison with the findings of acute anterior myocardial infarction. AB - The clinical course of 'Takotsubo' cardiomyopathy closely resembles that of acute myocardial infarction (AMI) and coronary angiography (CAG) is usually performed to distinguish the 2 conditions during the acute phase. The present study was designed to determine whether the standard 12-lead electrocardiogram (ECG) findings could help to distinguish 'Takotsubo' cardiomyopathy from anterior AMI. The study group comprised 13 patients with 'Takotsubo' cardiomyopathy and 13 consecutive patients with anterior AMI. Patients with 'Takotsubo' cardiomyopathy had abnormal Q waves less frequently than patients with anterior AMI (15% vs 69%, p=0.008). No reciprocal changes were seen in the inferior leads in patients with 'Takotsubo' cardiomyopathy (p=0.0003). The ratio of ST-segment elevation in leads V(4-6) to V(1-3) (SigmaSTeV(4-6)/V(1-3)) was significantly higher in patients with 'Takotsubo' cardiomyopathy (1.55+/-0.53 vs 0.57+/-0.58, p=0.0004). The QTc interval was significantly longer in patients with 'Takotsubo' cardiomyopathy. The absence of reciprocal changes, absence of abnormal Q waves, and a SigmaSTeV(4 6)/V(1-3) >/=1 all showed a high sensitivity and specificity for diagnosing 'Takotsubo' cardiomyopathy. Furthermore, the combination of the absence of reciprocal changes and a SigmaSTeV(4-6)/V (1-3) >/=1 had a greater specificity (100%) and overall accuracy (91%) than either criteria. Therefore, the standard 12-lead ECG on admission can help to distinguish 'Takotsubo' cardiomyopathy from anterior AMI. PMID- 12890912 TI - Differential increase in natriuretic peptides in elite dynamic and static athletes. AB - The echocardiographic measures and plasma concentrations of either atrial natriuretic peptide (ANP) or brain natriuretic peptide (BNP) were compared in elite judo practitioners (static athletes), elite marathon runners (dynamic athletes) and healthy controls to investigate the relationship between the different types of left ventricular (LV) hypertrophy and plasma concentrations of natriuretic peptides in athletes. The LV mass and LV wall thickness of marathon runners and judo practitioners were significantly greater than those of controls. The LV end-diastolic dimension index was significantly larger in the marathon group, but smaller in the judo group. The left atrial dimension (LAD) index was significantly larger only in marathon runners. Plasma BNP concentrations were higher in both the judo and marathon groups than in controls, and positively correlated with LV mass as well as with deceleration time. Plasma ANP concentrations were significantly higher in marathon runners than in the controls and judo groups, and positively correlated with the LAD index, but negatively correlated with ejection fraction. Multivariate analyses showed that the type of athlete and LAD index were independent predictors of plasma BNP and ANP concentrations, respectively. Thus, there is an intimate link between plasma concentrations of natriuretic peptides and cardiac morphology in different types of athletes. PMID- 12890913 TI - Timing of anti-platelet effect after oral aspirin administration in patients with sympathetic excitement. AB - Aspirin is used in percutaneous coronary interventions (PCI) for acute myocardial infarction (AMI) to prevent thrombosis. It is reported that the aspirin concentration in blood reaches its peak approximately 20 min after oral administration in healthy volunteers, but the absorption and bioavailability of aspirin in AMI may be quite different. In the present study patients undergoing coronary angiogram for the first time were enrolled as a model of sympathetic excitement and the timing of the antiplatelet effect after oral aspirin administration was investigated. Aspirin (162 mg) was administered to the patients in a catheter laboratory. Platelet count, aspirin concentration, and platelet aggregation were measured at scheduled timepoints before and up to 120 min. Ticlopidine was administered in the same procedure, and platelet count and platelet aggregation were evaluated at 0 and 120 min. There was no significant change in the platelet count. Aspirin concentration in blood had not reached its peak by 120 min. Platelet aggregation induced by collagen or ADP began to be inhibited 45 min after aspirin administration. No significant inhibition of platelet aggregation was observed up to 120 min following ticlopidine administration. During sympathetic excitement, aspirin absorption and its antiplatelet effect were significantly delayed in these patients. Ticlopidine did not show any antiplatelet effect by 120 min. For PCI performed in a patient with a high level of sympathetic excitement, aspirin should be administered at least 45 min before the first balloon dilatation. PMID- 12890914 TI - Effect of beta-blockers on insulin resistance in patients with dilated cardiomyopathy. AB - The aim of this study was to evaluate the effect of beta-blockers on insulin resistance in patients with dilated cardiomyopathy (DCM). A secondary aim was to determine the effect of this treatment on plasma concentrations of tumor necrosis factor-alpha (TNF-alpha) and to investigate the relationships between this adipocytokine and insulin resistance. Insulin resistance determined using the Homeostatic Model Assessment (HOMA), echocardiographic measurements and analysis of plasma TNF-alpha concentration were carried out in 47 patients with DCM without diabetes mellitus before and after 6 months of beta-blocker therapy. A reduction in left ventricular dimensions and an associated increase in ejection fraction occurred with beta-blocker. The treatment resulted in a significant decrease in insulin resistance (HOMA index: Baseline, 2.73+/-3.36 vs, Month 6, 1.58+/-1.33, p=0.0347). Beta-blockade was also associated with a decrease in plasma TNF-alpha concentration although no significant relationship between this change and the improvement in insulin resistance was observed. Beta-blocker therapy in patients with DCM improved not only cardiac function, but also insulin resistance. The mechanism of the change in insulin function remains unclear, but may be related to improvements in left ventricular function or an attenuation of the inhibitory effect of reduction in TNF-alpha on insulin signaling. PMID- 12890915 TI - Beneficial effects of angiotensin-converting enzyme inhibition on sarcoplasmic reticulum function in the failing heart of the Dahl rat. AB - Inhibition of angiotensin-converting enzyme (ACE) retards the process of myocardial remodeling and contractile dysfunction that leads to heart failure. However, the intracellular mechanisms by which ACE inhibition preserves myocardial contractility are largely unclear. Using a model of heart failure induced by hypertension in Dahl salt-sensitive (DS) rats, the mechanisms by which ACE inhibitors (ACEI) exert a beneficial effect on myocardial contractility were studied. Dahl salt-resistant (DR) rats, DS rats not given temocapril (DS/T-), and DS rats treated with temocapril (10 mg/kg per day from 10 to 17 weeks of age, DS/T+) were fed an 8% NaCl diet from 8 to 17 weeks of age (n=8, each group). Echocardiography, hemodynamic measurement, histology, contraction of isolated skinned papillary muscle, and Western blot analysis were carried out. At an elevated final blood pressure similar to that of the DS/T- rats, DS/T+ rats exhibited (1) a decrease in left ventricular (LV) mass associated with decreases in both cardiomyocyte size and interstitial fibrosis; (2) improvement of both systolic and diastolic LV function; and (3) an increase in caffeine contraction after constant Ca(2+)-loading with 8-bromo-cAMP into the sarcoplasmic reticulum (SR) associated with an increase in Ser16-phosphorylated phospholamban, as compared with the DS/T- rats. In addition to inhibition of myocardial remodeling, a restoration of the Ca(2+)-handling ability of the SR by normalized phosphorylated phospholamban may contribute to the improved LV contractile function achieved by chronic treatment with an ACEI. PMID- 12890916 TI - Nifekalant hydrochloride, a novel class III antiarrhythmic agent, suppressed postoperative recurrent ventricular tachycardia in a patient undergoing coronary artery bypass grafting and the Dor approach. AB - A patient with 3-vessel coronary artery disease and left ventricular aneurysm underwent coronary artery bypass grafting combined with the Dor approach. Five days later, ventricular tachycardia following short-coupled ventricular premature contractions suddenly occurred and was not responsive to class IB drugs (lidocaine and mexiletine), requiring frequent electrical cardioversion. After the administration of a novel class III drug, nifekalant hydrochloride, this electrical storm of ventricular tachycardia was completely suppressed together with the disappearance of ventricular premature contractions. Nifekalant hydrochloride (MS-551), a pure K(+) channel blocker, might be effective for postoperative recurrent ventricular tachyarrhythmias that are refractory to other antiarrhythmic agents. PMID- 12890917 TI - Combined atriofascicular and fasciculoventricular connections. AB - A 32-year-old man with a 20-year history of palpitation showed a short PQ interval and delta wave on 12-lead ECG. An electrophysiological study showed that the atrio-His (AH) and His-ventricular intervals were not influenced by rapid right atrial pacing or by bolus injection of adenosine triphosphate (ATP). The AH interval was shortened by coronary sinus ostium (CSos) pacing. Ventricular stimulus to the atrial activation interval was not affected by ATP but shortened by para-Hisian pacing and orthodromic His bundle activation was followed by atrial activation. The tricuspid - inferior vena cava isthmus was blocked by CSos pacing, but Ith conduction existed during low lateral right atrial pacing. PMID- 12890918 TI - Usefulness of a second temporary vena cava filter for preventing acute pulmonary thromboembolism. AB - The usefulness of a temporary inferior vena cava filter (t-IVC-f) for the prevention of acute pulmonary thromboembolism (APTE) associated with venous thromboembolism (VTE) has been established, but not the requirement for an additional therapeutic method when the t-IVC-f has captured a thrombus. A woman underwent implantation of a t-IVC-f just caudal to the bifurcation of the renal vein immediately before cesarean section (C/S) for VTE that had occurred during the third trimester of pregnancy. After receiving a combination of anticoagulant and fibrinolytic therapies after the C/S, the t-IVC-f was found to contain a thrombus and in order to prevent its dissemination during the removal of the closed filter, another filter was inserted and expanded proximal to the first. Insertion of an additional t-IVC-f may prevent occurrence of APTE during removal of the primary filter containing a friable thrombus after fibrinolytic therapy, and the technique proved useful in the present case of VTE during pregnancy. PMID- 12890919 TI - 'Tako-Tsubo cardiomyopathy' associated with syndrome malin: reversible left ventricular dysfunction. AB - A 66-year-old man developed a fever and had a syncopal attack during treatment with imipramine and amantadine for depression and Parkinson's disease. His muscular enzyme levels were very high, so he was diagnosed with incomplete syndrome malin and given hydration therapy. The electrocardiogram recorded an ST segment elevation like acute myocardial infarction in most leads, and the echocardiogram revealed left ventricular dysfunction with severe hypokinesis to dyskinesis of the anterior and apical wall regions, and hyperkinesis of the basal wall. One month from onset, the left ventricular contractility had not changed despite normal coronary arteries. Thallium-201((201)Tl) myocardial scintigraphy showed a perfusion defect and there was no accumulation of iodine-123((123)I) metaiodobenzylguanidine (MIBG) in the entire apex of the heart. Left ventricular function returned to normal and repeat (201)Tl scintigraphy showed recovery by the 4th month. However, there was still an absence of cardiac MIBG uptake. There are a number of reports from Japan of a syndrome demonstrating such reversible left ventricular dysfunction, called 'tako-tsubo cardiomyopathy', but the present case is the first to be associated with syndrome malin. A coronary microvascular abnormality and cardiac sympathetic denervation probably both play an important role in tako-tsubo cardiomyopathy. PMID- 12890920 TI - Loss of intracellular dystrophin: a potential mechanism for myocardial reperfusion injury. AB - Because the absence of sarcolemmal dystrophin renders cardiomyocytes vulnerable to mechanical force, the present study investigated whether sarcolemmal membrane fragility upon reperfusion is associated with the loss of membrane dystrophin. Dystrophin was distributed exclusively in the sarcolemmal membrane of buffer perfused rat cardiomyocytes, but was translocated to the myofibrils during 30 min of ischemia and then lost during reperfusion. Upon reperfusion, the membrane impermeable dye, Evans blue (EB), accumulated in cardiomyocytes depleted of dystrophin. Reperfusion with the contractile blocker 2,3-butanedione monoxime (BDM) resulted in no accumulation of EB in cardiomyocytes despite the loss of dystrophin. Upon withdrawal of BDM, however, EB accumulated in dystrophin depleted cardiomyocytes. Loss of sarcolemmal dystrophin may be involved in the mechanism of contractile force-induced reperfusion injury. PMID- 12890921 TI - Mapping quantitative trait loci using multiple phenotypes in general pedigrees. AB - The use of correlated phenotypes may dramatically increase the power to detect the underlying quantitative trait loci (QTLs). Current approaches for multiple phenotypes include regression-based methods, the multivariate variance of components method, factor analysis and structural equations. Issues with these methods include: 1) They are computation intensive and are subject to problems of optimization algorithms; 2) Existing claims on the asymptotic distribution of the likelihood ratio statistic for the multivariate variance of components method are contradictory and erroneous; 3) The dimension reduction of the parameter space under the null hypothesis, a phenomenon that is unique to multivariate analyses, makes the asymptotic distribution of the likelihood ratio statistic more complicated than expected. In this article, three cases of varying complexity are considered. For each case, the efficient score statistic, which is asympotically equivalent to the likelihood ratio statistic, is derived, so is its asymptotic distribution [correction]. These methods are straightforward to calculate. Finite sample properties of these score statistics are studied through extensive simulations. These score statistics are for use with general pedigrees. PMID- 12890922 TI - Linkage analysis of quantitative trait loci in the presence of heterogeneity. AB - Variance component modeling for linkage analysis of quantitative traits is a powerful tool for detecting and locating genes affecting a trait of interest, but the presence of genetic heterogeneity will decrease the power of a linkage study and may even give biased estimates of the location of the quantitative trait loci. Many complex diseases are believed to be influenced by multiple genes and therefore genetic heterogeneity is likely to be present for many real applications of linkage analysis. We consider a mixture of multivariate normals to model locus heterogeneity by allowing only a proportion of the sampled pedigrees to segregate trait-influencing allele(s) at a specific locus. However, for mixtures of normals the classical asymptotic distribution theory of the maximum likelihood estimates does not hold, so tests of linkage and/or heterogeneity are evaluated using resampling methods. It is shown that allowing for genetic heterogeneity leads to an increase in power to detect linkage. This increase is more prominent when the genetic effect of the locus is small or when the percentage of pedigrees not segregating trait-influencing allele(s) at the locus is high. PMID- 12890924 TI - Number of SNPS loci needed to detect population structure. AB - The study of the association of polymorphic genetic markers with common diseases is one of the most powerful tools in modern genetics. Interest in single nucleotide polymorphisms (SNPs) has steadily grown over the last decade. SNPs are currently the most developed markers in the human genome because they have a number of advantages over other marker types. One of the critical problems responsible for 'spurious' association findings in case-control studies is population stratification. There are many statistical approaches developed for detecting population heterogeneity. However the power to detect population structure by known methods is highly dependent on the number of loci utilised. We performed an analysis of SNPs data available in the public domain from The Single Nucleotide Consortia Ltd. (TSCL). Three populations, Afro-American, Asian and Caucasian, were compared. Estimation of the minimum number of SNPs loci necessary for detection of the population structure was performed. Two clustering approaches, distance-based and model-based, were compared. The model-based approach was superior when compared with the distance-based method. We found more than 65 random SNPs loci are required for identifying distinct geographically separated populations. Increasing the number of markers to over 100 raises the probability of correct assignment of a particular individual to an origin group to over 90%, even with conventional clustering methods. PMID- 12890923 TI - Choosing haplotype-tagging SNPS based on unphased genotype data using a preliminary sample of unrelated subjects with an example from the Multiethnic Cohort Study. AB - We describe an approach for picking haplotype-tagging single nucleotide polymorphisms (htSNPs) that is presently being taken in two large nested case control studies within a multiethnic cohort (MEC), which are engaged in a search for associations between risk of prostate and breast cancer and common genetic variations in candidate genes. Based on a preliminary sample of 70 control subjects chosen at random from each of the 5 ethnic groups in the MEC we estimate haplotype frequencies using a variant of the Excoffier-Slatkin E-M algorithm after genotyping a high density of SNPs selected every 3-5 kb in and surrounding a candidate gene. In order to evaluate the performance of a candidate set of htSNPS (which will be genotyped in the much larger case-control sample) we treat the haplotype frequencies estimate above as known, and carry out a formal calculation of the uncertainty of the number of copies of common haplotypes carried by an individual, summarizing this calculation as a coefficient of determination, R2h. A candidate set of htSNPS of a given size is chosen so as to maximize the minimum value of R2h over the common haplotypes, h. PMID- 12890925 TI - Pleiotropy and heterogeneity in the expression of atherogenic lipoproteins: the IRAS Family Study. AB - OBJECTIVE: Dyslipidemia is an important determinant of coronary disease. Phenotypic correlations between atherogenic lipids are well established, but the contribution of common genetic influences is less clear. METHODS: This study investigates the pair-wise genetic (rhog) and environmental (rhoe) correlations between apoB, LDL-C, HDL-C, and triglyceride (Tg) from Hispanic and African American families of the IRAS Family Study. RESULTS: Heritability estimates (h2) indicate significant genetic effects on apoB (h2=0.46+/-0.05), LDL-C (h2=0.40+/ 0.05), HDL-C (h2=0.47+/-0.05), and Tg (h2=0.35+/-0.05) (all p<0.001). Genetic and environmental correlations were strong for apoB-LDL-C (rhog=0.87, rhoe=0.84), apoB-Tg (rhog=0.38, rhoe=0.65), and HDL-C-Tg (rhog=-0.42, rhoe=-0.46). Environmental correlations were strong for apoB-HDL-C (rhoe=-0.40), LDL-C-HDL-C (rhoe=-0.24), and Tg-LDL-C (rhoe=0.33) with weak genetic correlations for these pairs (rhog=-0.09, 0.10, 0.09 respectively). CONCLUSIONS: These results suggest multiple pathways leading to atherogenic dyslipidemia. There are common genetic and environmental influences contributing to variations in apoB and LDL-C as well as apoB and Tg. In addition, the inverse relation between Tg and HDL-C appears to have both genetic and environmental basis. Identifying genes involved in atherogenic dyslipidemia will require careful dissection of the genetic architecture of these pathways. PMID- 12890926 TI - Sex-specific mutation rates for x-linked disorders: estimation and application. AB - Emerging human molecular data are adding to our knowledge about the frequency and pattern of genetic mutations. This not only gives important insight into the biological processes underlying mutation, but also provides data which must be incorporated in the clinical setting. An example is the assumption of equal mutation probability in the male and female germ lines. This is a key assumption in Bayesian risk calculation for families segregating an X-linked recessive disorder. For some disorders, data are now available that demonstrate that the mutation probability in males differs from that in females. In this paper, we review the estimation of the male-female mutation rate ratio, including the construction of confidence intervals, and apply sex-specific mutation rates to carrier risk calculation in a variety of pedigree structures. In several instances, the difference in risk is substantial. PMID- 12890928 TI - FLJ14813 missense mutation: a candidate for autosomal dominant thrombocytopenia on human chromosome 10. AB - The gene for a novel nonsyndromic autosomal dominant thrombocytopenia has been previously mapped to a region on human chromosome 10p11-12 (THC2, OMIM number *188000). This disorder is characterized by moderate thrombocytopenia and incomplete differentiation of megakaryocytes. We report here a novel missense mutation in the human gene FLJ14813 that segregates perfectly with thrombocytopenia in our kindred of 51 family members. The mutation is not detected in 94 random unrelated and unaffected individuals, nor is it reported in the Entrez single nucleotide polymorphism (SNP) database. A substitution of cytosine for guanidine (G to C) at nucleotide position 565 was present in all thrombocytopenic family members, causing a predicted substitution of aspartic acid for glutamic acid (E167D) in exon four. PMID- 12890927 TI - Estimation and tests of haplotype-environment interaction when linkage phase is ambiguous. AB - In the study of complex traits, the utility of linkage analysis and single marker association tests can be limited for researchers attempting to elucidate the complex interplay between a gene and environmental covariates. For these purposes, tests of gene-environment interactions are needed. In addition, recent studies have indicated that haplotypes, which are specific combinations of nucleotides on the same chromosome, may be more suitable as the unit of analysis for statistical tests than single genetic markers. The difficulty with this approach is that, in standard laboratory genotyping, haplotypes are often not directly observable. Instead, unphased marker phenotypes are collected. In this article, we present a method for estimating and testing haplotype-environment interactions when linkage phase is potentially ambiguous. The method builds on the work of Schaid et al. [2002] and is applicable to any trait that can be placed in the generalized linear model framework. Simulations were run to illustrate the salient features of the method. In addition, the method was used to test for haplotype-smoking exposure interaction with data from the Childhood Asthma Management Program. PMID- 12890929 TI - Localization of a novel autosomal recessive non-syndromic hearing impairment locus (DFNB38) to 6q26-q27 in a consanguineous kindred from Pakistan. AB - For autosomal recessive nonsyndromic hearing impairment over 30 loci have been mapped and 19 genes have been identified. DFNB38, a novel locus for autosomal recessive nonsyndromic hearing impairment, was localized in a consanguineous Pakistani kindred to 6q26-q27. The affected family members present with profound prelingual sensorineural hearing impairment and use sign language for communications. Linkage was established to microsatellite markers located on chromosome 6q26-q27 (Multipoint lod score 3.6). The genetic region for DFNB38 spans 10.1 cM according to the Marshfield genetic map and is bounded by markers D6S980 and D6S1719. This genetic region corresponds to 3.4 MB on the sequence based physical map. PMID- 12890930 TI - Peritoneal fluid concentrations of ciliary neurotrophic factor, a gp130 cytokine, in women with endometriosis. AB - Ciliary neurotrophic factor (CNTF) is a gp130 neuroregulatory cytokine belonging to the interleukin-6-type cytokine superfamily. Several lines of evidence suggest that the concentrations of interleukin-6 are elevated in the peritoneal fluid of endometriosis patients. To our knowledge, no study on whether this might also be the case for CNTF levels has been published previously. The CNTF concentrations in the peritoneal fluid were measured by ELISA in 51 women with (n = 31) and without (n = 20) endometriosis. Surgery was scheduled during the proliferative or the secretory phase of the menstrual cycle. CNTF was detectable in the peritoneal fluid of 43% of the women tested. The concentrations of CNTF showed no correlation with the presence of endometriosis or the phase of the menstrual cycle. We found no evidence to suggest that CNTF is involved in the pathogenesis of pelvic endometriosis. PMID- 12890931 TI - Nutritionally regulated hormonal factors in prolonged postnatal growth retardation and its associated adverse neurodevelopmental outcome in extreme prematurity. AB - Recent published data show that at hospital discharge, most infants born at <30 weeks of gestation would not achieve the median birth weight of the reference fetus at the same postconceptional age, and many would be less than the 10th centile. Estimating from the current recommendations of calorie and protein intakes, these infants accrue large deficits in intakes of protein and calorie during the first weeks of life. Postnatal growth retardation over a prolonged period of time is related to neurodevelopmental delays. While a total energy intake of 120 kcal/kg/day has generally been considered adequate, protein requirement in low gestation infants remains a matter for debate. Increasing the dietary protein:calorie ratio has previously been proposed as a strategy to enhance growth and to achieve a body composition similar to that of the reference fetus. Previous study data reveal that serum insulin-like growth factor I (IGF-I) concentration is positively correlated with protein intake, and nitrogen retention, in turn, is positively correlated with serum IGF-I concentration. Remarkably, elevated serum growth hormone but low serum IGF-I concentrations have been reported in low gestation infants and in infants with intrauterine growth retardation, suggesting IGF-I being a nutritionally regulated hormonal factor in the postnatal growth retardation. As neurodevelopment in extreme prematurity is likely affected by multiple factors, we hypothesize that a combined strategy of the previously proposed hormonal supplement with hydrocortisone and tri iodothyronine together with increased dietary protein intake (progressively increasing from 1.5 g/kg/day intravenously administered amino acids immediately after birth, then 3.6 g/100 kcal at approximately 125 kcal/kg/day when enterally fed till the infant reaches a body weight of >or=1.8 kg and at >or=50th centile weight of the reference fetus at the same postconceptional age) would likely be synergistic and more effective in improving neurodevelopmental outcome. PMID- 12890932 TI - Effects of antenatal antibiotics on the incidence and bacteriological profile of early-onset neonatal sepsis. A retrospective study over five years. AB - BACKGROUND: Recommendations for the use of antenatal antibiotics have been widely implemented in the past few years, notably to prevent group B streptococcal disease or to prolong pregnancy in the case of preterm premature rupture of the membranes. OBJECTIVES: We designed a retrospective study to assess the potential effects of this increasing use of antibiotics on the incidence and bacteriological profile of early-onset neonatal sepsis (EONS). METHODS: All neonates referred to our department for suspected EONS from January 1 1995 through December 31 1999 were included. Antenatal antibiotic exposure together with clinical and microbiological data from the neonatal period were gathered and analyzed on a yearly basis. RESULTS: Of the 485 newborns who met the inclusion criteria, there were 101 cases of culture-confirmed sepsis; 339 cases of suspected sepsis and 69 cases of confirmed sepsis involved children born in the hospital, among a total of 16,627 live births registered in our center over the study period. The overall incidence of EONS dropped from 6.8 to 0.6/1,000 births between 1995 and 1999 (p < 0.001), but the rate of group B streptococcal infection decreased much more rapidly than that of non-group B streptococcal infection. We observed a trend towards the emergence of ampicillin-resistant Escherichia coli strains, which were isolated in seven cases. Among E. COLI infections, ampicillin resistance was statistically linked with antenatal antibiotic use (p = 0.025). We also delineated several risk factors associated with these infections. CONCLUSION: In our center, antenatal antibiotic treatment was effective in reducing the incidence of EONS, but this benefit may come at the cost of favoring the emergence of ampicillin-resistant organisms causing severe neonatal infections. Antenatal and postnatal antibiotic treatment strategies should take this adverse effect into account. PMID- 12890933 TI - Predictive power of initial severity of pulmonary disease for subsequent development of bronchopulmonary dysplasia. AB - Using retrospectively acquired data from 138 mechanically ventilated premature infants, logistic regression was used to determine the relationships between the risk of bronchopulmonary dysplasia (BPD) and indices of initial respiratory disease severity [oxygen index (OI) and alveolar-arterial pO(2) difference (A-a DO(2))]. Indices were calculated from the first arterial blood gas analysis after initial surfactant administration. Infants were also classified as having mild [OI <4 cm H(2)O x mm Hg(-1), A-a DO(2) <150 mm Hg (20 kPa)] or severe [OI >or=10 cm H(2)O x mm Hg(-1), A-a DO(2) >or=300 mm Hg (40 kPa)] respiratory disease, and the ability of this classification to predict subsequent BPD risk was calculated. OI and A-a DO(2) were significantly higher in the BPD group. Logistic regression analysis showed that BPD risk increased linearly with both OI (9%/cm H(2)O x mm Hg(-1)) and A-a DO(2) [16%/50 mm Hg (16%/6.7 kPa)]. However, the predictive power (receiver-operator characteristic) of these models was modest. Unexpectedly, 29% of infants with mild initial disease developed BPD. These data suggest that, while BPD prediction in infants with severe disease is straightforward, the identification of those few infants with mild to moderate disease destined to develop BPD remains problematic. PMID- 12890934 TI - Prevention of Candida colonization prevents infection in a neonatal unit. AB - This study represents a 1-year surveillance period using our epidemiology-based principles published and successfully followed since 1979: weekly culture for yeasts of oral and anal swabs, treatment with oral nystatin of all colonized newborns, and good hygiene/handwashing. Colonization was demonstrated in 23 out of 791 newborns admitted from October 1998 to September 1999. Twenty-two strains of Candida were identified: 16 C. albicans, 2 C. parapsilosis, 3 C. glabrata, and 1 C. tropicalis. Symptoms were erythema of the buttocks in 6 colonized newborns. No other culture positive for Candida could be found. Previous contamination was the main source (previous stay in an intensive care unit, rarely maternal origin). Contamination in the unit was unlikely. Eradication of Candida could be observed within 1 week. These good results, controversial in the literature, were obtained following epidemiological conclusions and support our guidelines. PMID- 12890935 TI - Blood volume assessment with hemoglobin subtype analysis in preterm infants. AB - The aim of the study was to assess plasma volume (PV) and red cell volume (RCV) in preterm infants with a liquid chromatographic hemoglobin subtype method (HbST) based on dilution of autologous fetal hemoglobin (HbF) by donor adult hemoglobin and measurement of the amount of HbF. PV determination with the indocyanine green dye dilution technique (ICG) was used as a reference method. Eight infants, median gestational age 29.8 weeks (range 27.6-30.9 weeks) and median birth weight 1,300 g (range 1,030-1,760 g), were studied at a median age of 3.0 days (range 1 6 days). RCV was 33.6 +/- 12 ml/kg measured with ICG and 32.1 +/- 5.2 ml/kg with HbST. PV was 47.0 +/- 18 and 40.2 +/- 6.6 ml/kg, respectively. There was a close correlation between the RCVs measured with the two techniques (Pearson correlation 0.83, p < 0.05). In conclusion, HbST provides a reliable and safe determination of RCV in preterm infants. PMID- 12890936 TI - Maternal diet with diverse omega-6/omega-3 ratio affects the brain docosahexaenoic acid content of growing chickens. AB - Eggs with diverse omega-6/omega-3 ratio produced by feeding breeder hens a wheat soybean meal-basal diet containing 5% (wt/wt) sunflower oil (H(omega)6), 5% fish oil (H(omega)3) or 2.5% sunflower oil plus 2.5% fish oil (M(omega)3omega6) were incubated. The hatched chicks were fed a docosahexaenoic acid (DHA)-deficient diet up to 6 weeks of age. The fatty acid composition of chick brain was determined on 0, 2, 4 and 6 weeks and brain weight was taken on day 0 and day 42. The omega-6/omega-3 ratios were 37.12, 4.21 and 0.98 for the maternal diet; 28.36, 2.83 and 0.89 for the egg yolk; 1.94, 0.48 and 0.18 for hatched chick brain (p < 0.05). At 2 weeks of age, the omega-6/omega-3 ratios were 1.88, 0.81 and 0.60 for chicks hatched from hens fed H(omega)6, M(omega)3omega6 and H(omega)3 diets, respectively. The brain DHA contents at 0 and 2 weeks of age were Homega3 > M(omega)3omega6 < H(omega)6 (p < 0.05) and at 4 and 6 weeks of age H(omega)3 = M(omega)3omega6 > H(omega)6. Dietary C18:3omega3 in the starter and finisher diet did not increase brain DHA (p > 0.05). The significant increase in the content of C22:5omega3 at 6 weeks of age in group 1 birds with a concomitant reduction in DHA suggests a weak delta-4 desaturation but an effective delta-6 and delta-5 desaturation similar to human infants. Considering the role of DHA in early brain development and growth, the maternal supply of DHA during growth might be of importance when fed a DHA-deficient neonatal diet. PMID- 12890937 TI - Positive end-expiratory pressure modulates perfluorochemical evaporation from the lungs. AB - To study the effects of positive end-expiratory pressure (PEEP) level on perfluorochemical (PFC) elimination profiles (E(L)), 6 ml/kg of perflubron were instilled into healthy anesthetized rabbits. The ventilation strategy was to maintain constant minute ventilation (300 ml/kg/min) and mean airway pressure (7 8 cm H(2)O) while randomly changing the PEEP levels from 5 to 0, 1, 3, and 10 cm H(2)O, each for a period of 15 min. The PFC content in the expired gas was measured and the E(L) was calculated. There was a significant reduction in the E(L) when decreasing the PEEP levels from 5 to 0 cm H(2)O, but no differences were seen when the PEEP was increased from 5 to 10 cm H(2)O. The results indicate that PEEP levels influence PFC elimination profiles; therefore, the measurement of the E(L) and PEEP levels should be considered when optimizing supplemental PFCs during partial liquid ventilation. PMID- 12890938 TI - Ontogenetic expression of estrogen and progesterone receptors in the mouse lung. AB - In the mammalian lung, estrogen and progesterone seem to be import for the morphological and functional maturation. Despite this, only sparse information is available on the onset and ontogenic expression of estrogen receptors (ER) and progesterone receptors (PR) in the perinatal lung. The expression patterns of the two known nuclear isoforms ER-alpha and ER-beta and the PR were analyzed in the pre- and postnatal lungs of BALB/c mice. Steroid receptor mRNA expression of all three receptors were highest in the prenatal lung and declined thereafter to significantly lower levels in the postnatal and adult lung. The ontogenetic pattern of ER and PR expression supports the view that both gonadal steroids are pivotal for prenatal lung maturation and development. PMID- 12890939 TI - Prolonged mechanical ventilation induces pulmonary inflammation in preterm infants. AB - Lung inflammation plays an important role in the pathogenesis of chronic lung disease in preterm infants. To test the hypothesis that prolonged mechanical ventilation induces pulmonary inflammation, we analyzed pro- and anti inflammatory mediators in bronchoalveolar lavage fluid obtained from ventilated preterm infants having respiratory distress syndrome. Our results show a strong correlation between the duration of mechanical ventilation and the amount of proinflammatory mediators. However, the anti-inflammatory cytokine interleukin 10 remained stable during the whole period of mechanical ventilation. These data support the hypothesis that prolonged mechanical ventilation contributes to the development of chronic lung disease by the induction of lung inflammation without adequate stimulation of the counterregulatory cytokine interleukin 10 in preterm infants with respiratory distress syndrome. PMID- 12890940 TI - Maternal and umbilical cord serum leptin concentrations in small-for-gestational age and in appropriate-for-gestational-age neonates: a maternal, fetal, or placental contribution? AB - Leptin is secreted during pregnancy by the placenta and by the maternal and fetal adipose tissues. The leptin levels mainly reflect the amount of fat stored and thus are indicative of the energy balance, i.e., small-for-gestational-age (SGA) neonates represent the negative metabolic balance of in utero starved babies. We chose to compare maternal and umbilical cord leptin levels in pregnancies complicated by asymmetrical SGA versus those with appropriate-for-gestational-age (AGA) neonates as well as a model of multifetal growth concordant gestations in order to establish through the 'leptin link' the relative contributions of mother, fetus, and placenta to fetal weight. We found that the maternal leptin levels at delivery correlated poorly with the maternal weight gain/body mass index and with neonatal birth weight. Furthermore, the umbilical cord leptin levels correlated well with neonatal and placental weights in the AGA group but not in the SGA group. As in AGA singleton pregnancies, in multifetal uncomplicated pregnancies, the umbilical cord leptin levels correlated well with the birth weight of individuals, regardless of the status of the twin or triplet in the set. Thus, we speculated that in SGA neonates the birth weight represents the lean body weight and the low adipose tissue content (as opposed to the AGA neonates who have a substantial adipose tissue content) and, therefore, reflects mainly the basic placental contribution. PMID- 12890941 TI - Resuscitation of premature infants: what are we doing wrong and can we do better? AB - Neonatal resuscitation is based on experience with little evidence to support the methods advocated. Current guidelines make no distinction between the techniques for term and very premature infants. The guidelines support the use of 100%, cold, dry oxygen delivered with devices that provide variable peak inspiratory pressures and tidal volumes with no positive end-expiratory pressure (PEEP). It is possible that these techniques damage the lungs. Self-inflating resuscitation bags give no indication about leaks, produce variable inflating pressures, do not provide PEEP and cannot deliver prolonged inflations. Flow-inflating bags will not work if there is leak at the facemask and also have variable inflating pressures. Although they can provide PEEP and deliver prolonged inflations, they require considerable skill to use. The Neopuff is relatively easy to use, provides PEEP and steady inflating pressure and does not achieve the set pressures if there is a mask leak. Continuous positive airway pressure and PEEP are used in the neonatal intensive care unit to maintain lung volume. It is surprising they are not routinely recommended for resuscitation when establishing the lung volume is paramount. Volutrauma is a potential problem in neonatal resuscitation and yet none of the devices give any indication of the tidal volume delivered. There is considerable potential for improvement in techniques of neonatal resuscitation through the application of evidence already available and much scope for further research in this field. PMID- 12890942 TI - Genetic control of lung development. AB - Lung organogenesis is a developmental process that starts in human 4-5 weeks after conception and continues during the first years of life. It can be subdivided in six different stages: embryonic, pseudoglandular, canalicular, saccular and alveolar stage and stage of vascular maturation. In each of these periods, multiple molecules like transcription factors, growth factors and other signaling molecules and their respective receptors control and coordinate the course of events by a distinct expression and activity over space and time. Epithelial-mesenchymal interactions, physiological mechanical forces as well as humoral factors modulate some of these expression patterns. Although numerous key players and their mode of action have been discovered, many wait to be unveiled. Herein, we will summarize the current concepts of lung development with special consideration of the genetic control of lung genesis, growth and maturation. PMID- 12890943 TI - Interaction between surfactant and ventilatory support in newborns with primary surfactant deficiency. AB - The interaction between exogenous surfactant and various modes of ventilatory support in terms of timing, quality and quantity can influence both short- and long-term outcomes of immature infants. Alterations to the pulmonary surfactant system can occur with all forms of mechanical ventilation. Experimental data suggest possible interaction between ventilatory support and exogenous surfactant even during the first breaths in the delivery room. The adverse effect on surfactant function at this time can increase the need for and duration of ventilatory support. The logical approach to ventilatory support is to be minimally aggressive with optimal recruitment of the lungs to avoid ventilator induced lung injury. Nasal continuous positive airway pressure (CPAP) in combination with early prophylactic surfactant administration may be an effective and less damaging method capable of reducing the need of artificial ventilation, but its benefit has not been proven in extremely preterm infants less than 28 weeks' gestation. Because of unproven efficacy of nasal CPAP in extremely premature infants, the population most at risk for adverse pulmonary and neurologic outcome, this paper focuses on the comparison of conventional and high frequency oscillatory ventilation (HFOV) with respect to alteration of surfactant function, and short- and long-term outcomes, in both human and experimental trials. Though the two most recent large clinical trials provide reassurance with respect to the safety of first-intention high-frequency ventilation, the reduction in the risk of chronic lung disease appears to be only modest or absent. Recent laboratory investigations suggest that the key element of HFOV, namely optimization of volume, can, under some circumstances, be replicated with low tidal volume conventional ventilation and high positive end-expiratory pressure. Recent introduction of patient-triggered volume-targeted conventional ventilation into clinical practice offers the promise of a practical means of providing gentle conventional ventilation capable of minimizing ventilator induced lung injury. Ultimately, well-designed comparative clinical trials with long-term outcomes are essential to accurately quantify risks and benefits of any new approach to mechanical ventilation. Without such data, these experimental results should not be extrapolated into clinical practice, because of the multifactorial pathophysiology of the development of chronic pulmonary disease in extremely premature infants and the risk of unanticipated adverse effects. PMID- 12890944 TI - Current dilemmas in postnatal steroid therapy for chronic lung disease in preterm infants. AB - Postnatal steroid therapy has been widely used for the prevention and treatment of chronic lung disease in preterm infants. Recently, evidence has accumulated to suggest that the risks of therapy may outweigh the possible benefits. In 2002, the American Academy of Pediatrics and the Canadian Paediatric Society issued guidelines for steroid use. While clarifying a confusing situation, these guidelines have given rise to much controversy. This review will summarize current information on the clinical trials of steroids and also consider the issues surrounding the guidelines. PMID- 12890945 TI - Impaired alveolar development and abnormal lung elastin in preterm lambs with chronic lung injury: potential benefits of retinol treatment. PMID- 12890946 TI - Trials of nitric oxide. PMID- 12890947 TI - Lymph node staging in clinically localized prostate cancer. PMID- 12890948 TI - Port-site metastases: fact or fiction? PMID- 12890949 TI - Relook percutaneous nephrolithotomy: a simple technique to re-enter the pelvicalyceal system. AB - We describe a simple technique of reentry into the pelvicalyceal system for removal of residual fragments that is simple, rapid and cheap. PMID- 12890950 TI - Natural killer T cells in patients with prostatic carcinoma. AB - Recent interest has focused on natural killer T (NKT) cells as effector cells in a possible new form of immunotherapy. NKT cells express T cell and natural killer cell receptors. They are CD1d-dependent and can be stimulated by alpha galactosylceramide. We have studied peripheral blood levels of NKT cells simultaneously with CD8+ cells and natural killer cells in 54 patients with prostatic carcinoma. Levels of NKT cells in patients with androgen withdrawal and stable PSA levels were 211.8 +/- 138.4 cells/microl and in patients with rising PSA levels were 27.0 +/- 22.1 cells/microl. A similar but less pronounced difference in CD8+ cells between these groups was found (656.4 vs. 212.0 cells/microl). Comparing all 54 patients studied, a positive correlation was found between the levels of NKT cells and CD8+ cells (r = 0.83). Our results suggest an important and regulatory role of NKT cells in patients with prostatic carcinoma. PMID- 12890951 TI - Detection of prostate cancer and changes in prostate-specific antigen (PSA) six months after surgery for benign prostatic hyperplasia in patients with elevated PSA. AB - OBJECTIVE: To evaluate early postoperative results of patients with elevated prostate-specific antigen (PSA) levels who underwent surgery due to benign prostatic hyperplasia (BPH). PATIENTS AND METHODS: 64 patients who had lower urinary tract symptoms (LUTS), normal digital rectal examinations (DRE), elevated PSA levels and prostate biopsies reported as being benign pathologically in specimens obtained by transrectal ultrasound (TRUS)-guided biopsies, were included in the study. Patients were assessed in accordance with PSA density, free/total PSA ratio and uroflowmetric studies. Patients had no cancer pre- and postoperatively (according to operative specimens). Six months postoperatively, 32 patients were accepted for re-evaluation for all PSA parameters, routine tests and prostatic biopsies. RESULTS: 64 patients with a mean age of 66.8 (SD 6.72) were included in the study. Total PSA average value was 14.38 (SD 7.49) ng/ml. Free PSA average value was 2.11 (SD 1.43) ng/ml. Average PSA density and free/total PSA ratio were 25.19 SD (14.12) ng/ml/cm(3) and 14.53% (SD 5.35%) respectively. 56 patients had BPH, 7 had chronic prostatitis and 1 had prostatic intraepithelial neoplasia (PIN) preoperatively with biopsies. Re-biopsy of the patient with PIN was reported as BPH. In pathologic examination with resected tissues postoperatively, 49 patients had BPH, 14 had chronic prostatitis and 1 had PIN. In the sixth month, average values of free/total PSA were 0.45 (SD 0.26) and 3.71 (SD 4.96) ng/ml respectively. Average PSA density and free/total PSA ratio were 12.41 (SD 13.8) ng/ml/cm(3) and 19.59% (SD 10.33%) respectively. There were significant decreases in PSA densities (p < 0.001) and increases in free/total PSA ratios (p = 0.004). Seven patients still had elevated PSA levels 6 months postoperatively. Three of 7 patients were reported as chronic prostatitis. One of them was indicated as prostatic carcinoma who was reported as PIN preoperatively. All other patients were stated as BPH at re-biopsies. CONCLUSION: If pretreatment biopsies are negative and operative specimens are also benign in patients with high PSA values, these patients can be followed up like usual BPH patients, but long-term follow-up is still unclear. PMID- 12890952 TI - Prostate-specific antigen levels and density in the internal and external glands of the prostate in benign prostatic hyperplasia patients with normal or gray-zone PSA levels. AB - OBJECTIVE: In patients with benign prostatic hyperplasia (BPH) showing normal prostate-specific antigen (PSA) levels (normal PSA group) and those with BPH showing gray-zone PSA levels (gray-zone PSA group), we assessed PSA levels secreted from the internal and external glands. MATERIALS AND METHODS: We performed transurethral enucleation of the prostate (TUE) in 102 BPH patients with normal PSA and 59 BPH patients with gray-zone PSA at our department from 1999 to 2001. Preoperatively and approximately 6 months postoperatively, we measured serum PSA levels and determined prostatic volumes via transrectal ultrasonography (TRUS) to calculate PSA levels secreted from the internal and external glands as well as various PSA density (PSAD) values. RESULTS: The total PSA level was 1.8 and 6.1 ng/ml in the normal and gray-zone PSA groups, respectively. The PSA level of the external gland was 0.6 and 0.8 ng/ml and the PSAD of the external gland was 0.07 and 0.08 ng/ml/cm(3) in the normal and gray zone PSA groups, respectively. The internal gland PSA was 1.3 and 5.4 ng/ml and the internal gland PSAD value was 0.11 and 0.30 ng/ml/cm(3) in the normal and gray-zone PSA groups, respectively. CONCLUSIONS: Our results demonstrated that increased PSA levels in BPH cases with gray-zone PSA were attributable to increased PSA secreted from the internal gland rather than from the external gland. In our opinion, the determination of PSA and PSAD of the internal and external gland may be clinically significant in the future. PMID- 12890953 TI - Increased prostate-specific antigen in subclinical prostatitis: the role of aggressiveness and extension of inflammation. AB - OBJECTIVES: Subclinical prostatitis is a very frequent histologic finding in pathological examinations of prostate biopsy and prostate surgery material. In this study, we tried to investigate the correlation between the morphological parameters of histological prostatitis and total serum prostate-specific antigen (PSA)-PSA density (PSAD) to determine if either the extent or aggressiveness of inflammation might affect serum PSA. METHODS: 269 patients who had undergone TURP or transvesical prostatectomy with pathological diagnosis of BPH and prostatitis were included in the study. We retrospectively reviewed and scored the extent and aggressiveness of inflammation in prostate specimens of BPH, according to the scale that has been reported by Irani et al. and then correlated those scores with PSA and PSAD. RESULTS: When the inflammation grades correlated with PSA and PSAD, the extent of the inflammation did not show a significant correlation with total PSA and PSAD (p > 0.05). However, there was a statistically significant correlation between aggressiveness grades and total PSA and PSAD (p < 0.001). Median PSA levels in grades 0, 1 and 2 of aggressiveness of inflammation were 3.2, 4.2 and 5.8 respectively. CONCLUSION: Aggressiveness grade of the inflammation in subclinical prostatitis is the most important morphological factor that is responsible for PSA elevation. We believe that it should be a more accurate guide for the clinician if pathologists report on the aggressiveness grades of the inflammation, especially on initial prostate biopsies, in order to help for timing of the further biopsy. PMID- 12890954 TI - Periprostatic nerve blockade before transrectal ultrasound-guided prostate biopsy: the Ankara Numune experience. AB - OBJECTIVES: To assess the efficacy and morbidity of periprostatic local anesthesia before transrectal ultrasound-guided biopsy of the prostate. METHODS: From August 2001 to December 2001, 58 patients underwent transrectal ultrasound guided prostate biopsy at the 2nd Department of Urology, Ankara Numune Education and Research Hospital. Fifty patients who fulfilled the inclusion criteria were randomized into 2 groups of 25 patients each. Group 1 received periprostatic local anesthesia with 1% lidocaine while group 2 received no local anesthesia. Pain scale responses were analyzed for each aspect of the biopsy procedure with a visual analog scale. RESULTS: There was no difference between the 2 groups in pain scores during digital rectal examination, intramuscular injection and probe insertion. The mean pain scores during needle insertion in group 1 receiving periprostatic nerve block and in group 2 receiving no local anesthesia were 3.00 +/- 2.22 and 6.16 +/- 2.85, respectively, and were found to be significantly different (p < 0.001), but morbidity after the biopsy was not statistically different between the 2 groups. CONCLUSION: Periprostatic local anesthesia before prostate biopsy is a safe and easy method to increase patient comfort during the procedure. PMID- 12890955 TI - Patient-controlled epidural analgesia after major urologic surgeries. A comparison of tramadol with or without bupivacaine. AB - The efficiency and safety of patient-controlled epidural analgesia by using tramadol alone and combined with bupivacaine were investigated for postoperative pain treatment after major urological surgeries. For PCEA: in group I (n = 17) a loading dose of 20 mg tramadol with a continuous infusion of 1 mg/ml tramadol at a rate of 8 ml/h was given. In group II (n = 17), patients received an initial loading dose of 20 ml bupivacaine 0.125% and a supplemental continuous infusion of 8 ml/h. In group III (n = 17), a loading dose of 20 mg tramadol with 20 ml bupivacaine 0.125% were given and a supplemental infusion of 1 mg/ml tramadol in 20 ml bupivacaine 0.125% combination was begun with a rate of 8 ml/h. A demand epidural bolus dose of 5 ml with a lockout time of 30 min was also used in all patients. VAS for pain intensity, vital signs, sedation scale and side effects was monitored at 0, 15, 30 min and 1, 2, 3, 4, 8, 12, and 24 h of the postoperative period. Statistical significance was determined using Kruskal Wallis, Fisher's exact, analysis of variance for repeated measurements and Tukey tests. The hemodynamic values and sedation scales were insignificantly different (p > 0.05). The adequate analgesia was provided in all patients. However VAS values were significantly lower in group III than in groups I and II at every measurement (p < 0.05). The incidence of side effects in all three groups was low (p > 0.05). In conclusion, we suggested that a combination of tramadol with bupivacaine can provide the most effective and safe postoperative analgesia with minimal risk for side effects. PMID- 12890956 TI - How reliable is conventional urinary cytology in post-transplant patients? AB - INTRODUCTION AND OBJECTIVES: Factors like cold, flushing solutions, ischemia and reperfusion may alter the microscopic appearance of transitional cells leading to falsely positive results of urinary cytology in patients after kidney transplantation. After seeing 1 patient presenting with two consecutive highly suspicious cytology specimens 3 days after transplantation and no sign of urothelial tumor at retrograde urography, we analyzed the cytological picture of transitional cells in post-transplant patients. MATERIAL AND METHODS: We investigated 31 urine specimens of 11 patients undergoing kidney transplantation preoperatively (if possible) and on days 1, 3 and 9 postoperatively. Microscopic cytology was performed by using Papanicolaou's criteria: 0 - no cytology possible (no cells), I+II - negative cytology, III - doubtful, IV - suspicious for tumor, V - tumor cells. All microscopic examinations were performed by one experienced senior pathologist. RESULTS: Mean patient age was 55.8 (+/- 17.5) years, mean residual diuresis 856 (+/- 636) ml, mean cold ischemia time 13.6 (+/- 6.4) h, mean creatinine level on day 1: 582 microM/l, day 3: 533 microM/l and day 9: 259 microM/l. None of the urinary cytology results were suspicious for malignant transformation (Papanicolaou I+II). No patient presented signs of urothelial malignancy after a mean follow-up of 3 months. CONCLUSION: Although microscopic urinary cytology may be falsely positive in 1-12% of non-transplanted patients due to urothelia atypia, inflammation or radiation/chemotherapy, the present study suggests that conventional microscopic cytology examinations in post transplant patients are not heavily altered and do not lead to an increased false positive rate. PMID- 12890957 TI - Pyogenic psoas abscess: difficulty in early diagnosis. AB - AIM: To report on the clinical features, diagnosis, and treatment of psoas abscess (PA) with special attention to the presence of septic shock. PATIENTS AND METHODS: This study included 17 patients (mean age 66.2, range 43-81 years) with PA. Treatment consisted of intravenous administration of antibiotics and abscess drainage, either surgical or percutaneous with ultrasound guidance. RESULTS: The typical patients presented with fever >38 degrees C (16/17, 94%), pain in back, flank, or abdomen (15/17, 88%), hip flexion contracture with pain extension (14/17, 82%), and mass felt in the flank (5/17, 29%). All 8 patients without septic shock (100%) had the clinical triad (fever, pain in back, flank, or abdomen, and hip flexion contracture) as compared with 4 of 9 patients with septic shock (44%) (p = 0.012). The duration of symptoms before hospitalization was significantly shorter in the patients with septic shock (median 2, range 1-5 days) than in those without septic shock (median 18.5, range 11-63 days; (p = 0.0005). The mortality rates were 33% (3 of 9) and 0% (0 of 8) in the patients with and without septic shock, respectively (p = 0.071). CONCLUSIONS: PA patients with septic shock had a tendency to have nonspecific symptoms and an occult clinical course as compared with those without septic shock. A delay in diagnosis and treatment can result in a worse clinical outcome (death or totally disabled state). Increased awareness of this condition should lead to earlier diagnosis and treatment with improved outcomes. PMID- 12890958 TI - Surgical enucleation for the treatment of renal tumors. AB - INTRODUCTION: We analyzed our institutional experience with surgical enucleation for the primary treatment of small renal tumors. MATERIALS AND METHODS: Patient demographics, histological features, effect of different types of vascular control and outcome from surgery were analyzed in 45 patients. A majority of the tumors (67%) were diagnosed incidentally. RESULTS: All were stage T(1), 77% were low grade (I-II) and 23% were high grade (III-IV). Complication occurred in 12% of patients. At a mean follow-up of 34 months (range 7-97), 28 of 33 patients (84%) with malignant tumors were alive without evidence of disease. One patient with a solitary kidney developed recurrent tumor after enucleation that required nephrectomy. Mean operative time was significantly lower with the compression technique versus direct vascular control (164 +/- 12 min vs. 233 +/- 15 min, p = 0.002). There were no differences in outcome between the two techniques. CONCLUSIONS: Surgical enucleation is a safe and viable procedure for the treatment of small renal tumors. Manual compression of the kidney appears to be at least as effective as clamping of the renal vessels in obtaining vascular control during the procedure and is more expeditious. PMID- 12890959 TI - Outcome of nephron-sparing surgery: elective versus imperative indications. AB - INTRODUCTION: The increase in the detection of renal tumors incidentally in earlier stages has enhanced the enthusiasm for nephron-sparing surgery (NSS). PATIENTS AND METHODS: We performed NSS in 76 patients (53 male, 23 female) with a mean age 52.3 between December 1988 and September 2001. Patients were sub classified into 2 groups as elective or imperative indication group. They were compared regarding surgical technique, time of surgery, pathological analysis, complications, and disease free status. RESULTS: Elective indication group (group I) with a normal contralateral kidney consisted of 50 patients, whereas there were 26 patients in the imperative indication group (group II). Tumors were incidentally detected in 63%; 74% in group I and 42% in group II. Although the mean diameter of the tumor was slightly higher in group II (39.1 vs. 36.3 mm), this difference did not reach statistical significance (p > 0.05). The partial nephrectomy was performed more frequently compared to enucleation in group I (90 vs. 69%, p = 0.050). However, the mean operation time as well as the mean clamping time did not differ significantly between the two groups (p > 0.05). In the histological evaluation tumors were benign in 16 (21%) and malignant in 60 patients. All of the patients but one with renal cell carcinoma had stage T1-T2 disease. Major complications were observed in 14 (18%) and 12 were from group II. Complication rate was significantly higher in group II (p = 0.000). Of 60 patients with renal cell carcinoma, 2 died of unrelated causes. One patient died with multiple visceral metastases. One patient was lost to follow-up. In the remaining 56 patients with a mean follow-up of 37.1 months (1-152), local recurrence or distant metastases were not detected. Serum creatinine levels have remained almost the same compared to preoperative levels (1.2 +/- 0.6 vs. 1.5 +/- 0.9). Overall and cancer-specific survivals were 100 and 100% in group I, 85 and 95% in group II, and 94 and 98% for the entire patient population, respectively. CONCLUSION: NSS is an effective and reliable treatment in low stage renal tumors. It prevents unnecessary nephrectomy in benign lesions that could not be diagnosed preoperatively. However, the patients who underwent NSS with elective indication outcome with better results, compared to those with imperative indication. PMID- 12890960 TI - Renal cysts: can percutaneous ethanol injections be considered an alternative to surgery? AB - AIM: To evaluate the efficacy of a new sclerotization technique with pure ethanol in the treatment of symptomatic renal cysts. PATIENTS AND METHODS: Fourteen patients having renal cysts with a meant diameter of 10 (range 5-15) cm were treated. Our technique includes: ultrasound-guided percutaneous puncture with an 18-gauge needle, positioning of a 5-Fr catheter, complete cyst fluid aspiration, injection of pure alcohol equal to 15% of the initial cyst volume, and alcohol aspiration after 90 min. The procedure was repeated eight times within 5 days. The patients were followed up by ultrasound and/or CT scan for 1 year. RESULTS: All patients became symptom free. Follow-up showed a progressive reduction of the cyst diameter in all cases. Three cysts only (in 2 patients; cyst diameter <2 cm) persisted after 12 months. No significant complications were observed. CONCLUSIONS: In our experience, injections of pure ethanol in renal cysts, repeated after some days, were effective in eliminating recurrences and related symptoms. The procedure was not associated with significant complications. Our findings suggest that it be considered the first-choice procedure in the treatment of renal cysts, due to the good results and the low cost of ethanol. PMID- 12890961 TI - Open pyeloplasty in children: experience with an improved stenting technique. AB - OBJECTIVE: To describe the use of an alternative and improved nephrostostent in open pyeloplasty procedures. MATERIALS AND METHODS: A series of 27 consecutive patients (between 4 months and 14 years of age) undergoing an open pyeloplasty procedure had their urinary system drained via a nephrostostent which passes from the ureteric anastomosis to the skin and which unlike other described nephrostostents does not cross the vesico-ureteric junction. All children received an ultrasound and MAG3 scan 3 months post-operatively and were followed up for 18-24 months. RESULTS: The use of this stent was associated with no serious complications. There was a decreased incidence of vesico-ureteric reflux, post-operative urinary tract infection and stent displacement compared to previously described methods of drainage. CONCLUSION: This method of open pyeloplasty drainage, which has not been described before, could be associated with fewer complications than existing stenting techniques. PMID- 12890962 TI - Endopyeloureterotomy using the holmium:YAG laser for the management of adult ureteral and ureteropelvic junction obstructions. AB - AIM: To determine the efficacy of endopyeloureterotomy using a holmium:YAG laser for the management of adult benign ureteral and ureteropelvic junction obstructions. PATIENTS AND METHODS: We reviewed the clinical outcomes of eight primary procedures performed in 5 patients. The causes of the strictures were idiopathic in five and stone impaction in three procedures. All strictures except one were approached in a retrograde fashion with either a rigid (8.0 Fr) or flexible (9.3 Fr) ureteroscope. The only energy source employed was the laser, and no balloon dilation was performed. In all cases, a 4.8-Fr ureteral stent was left indwelling, and retrograde ureteropyelography was performed within 6-8 weeks to remove the catheter. RESULTS: Follow-up was performed in all patients for a mean period of 14.9 (minimum 10) months. Two strictures in 1 patient failed to be stent free, while six strictures were patent on postoperative radiographic imaging. There were no perioperative complications. CONCLUSIONS: Endopyeloureterotomy using a holmium:YAG laser is safe and effective. Its relative noninvasiveness and its retrograde approach make it a first-line alternative for the management of benign upper urinary tract strictures in adults. PMID- 12890963 TI - Low-dose low-osmolar urography with iohexol. AB - OBJECTIVES: To study the diagnostic efficacy of low-dose iohexol (90 mg iodine/kg body weight) for intravenous urography. MATERIALS AND METHODS: Intravenous urography was performed using low-dose iohexol in 50 patients with various urinary complaints. Patients with elevated blood urea and serum creatinine levels were excluded. The urographic films were evaluated with regard to diagnostic information about renal parenchyma, calyces, collecting system and overall image. Patients were also observed for adverse reactions. RESULTS: A satisfactory diagnosis could be reached in 94% of patients with low-dose iohexol. Only a minor reaction in the form of heat sensation was noted in 1 patient, which did not require any treatment. CONCLUSION: Iohexol is safe and very useful diagnostically in a low dose for routine intravenous urography. PMID- 12890964 TI - Gastrocystoplasty in pigs and effect of selective antral vagotomy on acid secretion of the reconstructed bladder. AB - INTRODUCTION: The aim of this study was to investigate the outcome of gastrocystoplasty and the effects of selective antral vagotomy (SAV) on the postprandial gastrin secretion from the antrum as well as on the acid secretion from the augmented bladder. MATERIALS AND METHODS: In this study on 12 male pigs, we applied subtotal cystectomy plus gastric augmentation plus SAV to the study group and the same procedure without SAV to the control group. The animals were followed up for 3 months with respect to feeding, weight, and urine output. The urine pH levels and the gastrin levels of the pigs in the two groups were then followed up and compared. RESULTS: The use of gastric segments in bladder reconstruction was found to be appropriate in terms of both gastric function and urinary system function. Nevertheless, regarding the effect of SAV, the differences between either the urinary pH levels or the gastrin levels of the pigs in the two groups were statistically significant. CONCLUSIONS: Although gastric segments in the bladder reconstruction were found to be appropriate in terms of both gastric function and urinary system function, SAV did not prevent postprandial gastrin secretion and the resulting increase of the urine acidity. PMID- 12890965 TI - Seromuscular gastrocystoplasty in dogs. AB - INTRODUCTION: The aim of this study was to investigate the feasibility of seromuscular gastrocystoplasty (SGCP) in an animal model and to compare it to conventional gastrocystoplasty (CGCP). MATERIALS AND METHODS: CGCP and SGCP (using gastric segments without mucosa) were each performed in 6 dogs. In both procedures, two-thirds of the dome of the bladder were excised and the gastric segment anastomosed to the bladder remnant. Cystography, cystomanometry, measurements of urine pH, and gross and microscopic pathological studies were carried out preoperatively, and postoperatively, at 6 and 12 weeks. RESULTS: All seromuscular gastric segments proved viable, and 6 weeks after the operation they were covered by a thin layer of transitional epithelium, which had gradually thickened by the end of the 12-week follow-up. There was no difference in bladder capacity and compliance between the two groups, however, fasting urinary pH values were higher (less acidic) in the SGCP group. CONCLUSIONS: Stripping off the mucosa of the gastric segment appears to stop hydrochloric acid secretion, thereby lessening the possible risk of ulceration, perforation, dysuria haematuria, metaplasia and malignancy. The uroepithelium overgrowth of the seromuscular gastric segments might provide a more physiological neo-bladder than when using full-thickness gastrocystoplasty. PMID- 12890966 TI - SRY-positive 46,XX male with cryptorchidism as the only presenting clinical feature. AB - The SRY gene, located on the short arm of the Y chromosome, is responsible for differentiation of the testis from the undifferentiated gonad. We report a 4-year old patient with male phenotype and female karyotype (46,XX) with cryptorchidism as the only presenting clinical abnormality. Fluorescent in situ hybridization analysis, using Y- and X-specific (whole chromosome painting WCP Y WCP X) DNA and SRY probes, detected a small Y chromosome fragment, including the SRY gene, transferred to the short arm of the X chromosome. PMID- 12890967 TI - Unexpected presentation of syringocele. Acontractile bladder. AB - Cowper's syringocele is an uncommon deformity in the male urethra that is a distention of the duct of the Cowper's gland. Symptoms attributed to Cowper's syringocele in adults consist of lower urinary tract symptoms. To our knowledge there are no data on acontractile bladder due to Cowper's syringocele. We describe a patient who had acontractile bladder and a Cowper's syringocele. PMID- 12890968 TI - Leiomyoma of the female urethra. A case report. AB - We report a case of leiomyoma of the female urethra and a review of the literature. A mass protruding from the urethral meatus caused voiding difficulty and dysuria. Uroflowmetry showed an obstructive pattern with low maximum urine flow. Cystourethroscopy revealed a mass arising from lateral urethral wall extending from meatus up to 2 cm proximally. Histopathological studies confirmed urethral leiomyoma. Surgery completely resolved the original symptoms. PMID- 12890969 TI - Infected hydrocoele of the canal of Nuck in a neonate. AB - Hydrocoele of the canal of Nuck is an uncommon presentation in the neonate and complications are rare. A 6-day-old girl presented with a right groin swelling from birth. The swelling remained static until age 3 days when it increased rapidly in size. There was no fever or gastrointestinal symptoms. Physical examination showed a large nontender mass extending from the right groin into the ipsilateral labium majus with distortion of the external genitalia. Needle aspiration yielded cloudy fluid, which gave a growth of Klebsiella on culture. A hydrocoele of the canal of Nuck was confirmed at surgery and was excised. Postoperative course was uneventful and there has been no recurrence at 2 years of follow-up. PMID- 12890970 TI - Clear cell adenocarcinoma of the bladder in a male patient: clinicopathologic analysis of a case. AB - Clear cell adenocarcinoma arising in the lower urinary tract is unusual. We report a case of clear cell adenocarcinoma of the bladder in a 54-year-old male. Cystoscopic examination revealed a tumour arising from the dome of the urinary bladder. Histologically, the tumour was chiefly composed of tubulocystic and papillary glands lined by glycogen-rich, cubical or hobnail cells with clear to eosinophilic cytoplasm. The tumour infiltrated into the inner muscular layer. The patient was treated by partial cystectomy. PMID- 12890971 TI - Benign retroperitoneal schwannoma presenting as a giant adrenal tumor. AB - We report a 51-year-old man with a primary adrenal tumor presenting as a giant juxta-adrenal benign schwannoma. The patient was initially admitted for persistent fevers and underwent a sepsis workup. Abdominal ultrasound initially revealed an incidental right suprarenal mass, which measured 10 x 10 x 11 cm, estimated by computed tomography. Following a successful laparotomy, histologic examination confirmed that this giant tumor was a benign retroperitoneal schwannoma. Postoperatively, the patient remains disease-free, undergoing rehabilitation. PMID- 12890972 TI - A false-negative diuretic radionuclide renography in a patient with urinary ascites due to retroperitoneal fibrosis. AB - In this case report we present a rare manifestation of a false-negative diuretic radionuclide renography in a patient with urinary ascites due to retroperitoneal fibrosis. Diuretic renography may lead to false-negative results and should be carefully evaluated in this group of patients. PMID- 12890973 TI - Placement of deep brain stimulators into the subthalamic nucleus or Globus pallidus internus: technical approach. AB - Deep brain stimulation (DBS) represents a major advance in the treatment of Parkinson's disease (PD). As more neurosurgeons enter this field, technical descriptions of implantation techniques are needed. Here we present our technical approach to subthalamic nucleus (STN) and globus pallidus internus (GPi) DBS implantation, based on 180 STN implants and 75 GPi implants. The essential steps in DBS implantation are magnetic resonance imaging (MRI)-guided stereotactic localization, confirmation of the motor territory of the target nucleus with microelectrode mapping, and intra-operative test stimulation to determine voltage thresholds for stimulation-induced adverse effects. Lead locations are documented by postoperative MRI in all cases. PMID- 12890974 TI - Motor cortex stimulation in the treatment of deafferentation pain. I. Localization of the motor cortex. AB - MRI and electrophysiological techniques to localize the primary motor cortex (MC) were performed on patients considered for MC stimulation for the treatment of deafferentation pain. The representation and trajectory of the rolandic fissure (RF) were accurately localized by external cranial landmarks and radiopaque fiducials superimposed on oblique MRI sections. In addition, the scalp distribution of the corticocortical responses elicited by acute epidural stimulation [motor cortex (MC) in frontal and sensory cortex (SC) in parietal scalp regions], and analgesic responses at the topographical representation of the painful periphery elicited by subacute epidural stimulation were found to be simple and reliable procedures to localize MC, SC and RF. PMID- 12890975 TI - Technical difficulties and perioperative complications of retrogasserian glycerol rhizotomy for trigeminal neuralgia. AB - In 139 patients, 260 consecutive retrogasserian glycerol rhizotomies for trigeminal neuralgia were retrospectively analyzed regarding technical surgical difficulties and immediate and early complications. Technical obstacles occurred in 47.3%. In 21 cases (8.1%), the surgical procedure had to be interrupted due to circumstances such as vasovagal reactions, cardiac arrest, or difficulties to find the trigeminal cistern. Complications or side effects, being either transient or persistent, occurred in 67.3%. In the vast majority, those unwanted effects were related to mild sensory deficits. However, in 28.1% the complications were other than mild affection of facial sensibility. These slightly graver complications included labial herpes (3.8%), anesthesia dolorosa (0.8%), moderate or severe affection of sensibility (18.8%), dysesthesia (22.7%), chemical meningitis (1.5%) and infectious meningitis (1.5%). In 5 patients (1.9%) hearing was affected. In one of them, this condition was also brought about by tinnitus, and in another patient a preexisting tinnitus deteriorated. Although the frequency of surgical difficulties was high, the success of the glycerol injection was hampered only in a minor number of procedures. The frequency of complications and side effects was high, but they were mostly mild due to their nature and non-disabling for the patient. However, long-lasting disabling side effects occurred, and this should not be neglected when informing patients preoperatively. The surgical training needed to perform the procedure is stressed, and the use of prophylactic antibiotics when accidentally penetrating the oral bucca is recommended. We consider retrogasserian glycerol rhizotomy to be a good surgical option for patients with trigeminal neuralgia not suitable for microvascular decompression and when pharmacological therapy is not sufficient or is not tolerated. PMID- 12890976 TI - Interventional MRI-guided frameless stereotaxy in pediatric patients. AB - INTRODUCTION: We prospectively reviewed our experience with intraoperative MRI (iMRI)-guided stereotactic procedures in pediatric patients. METHODS: All procedures were performed within the magnet bore of the General Electric Signa SP MRI system, which allows for either continuous real-time or periodic imaging. The internal optical tracking system was used to plan and monitor target localization and instrument trajectory. RESULTS: Fifteen patients underwent 16 frameless stereotactic procedures, consisting of 4 tumor biopsies and 12 cyst aspirations and stereotactic catheter placements (average age 6 years, range 6 weeks to 18 years). There were no hemorrhagic, neurologic or infectious complications. CONCLUSION: iMRI is an important component in expanding the horizon of minimally invasive neurosurgery for pediatric patients. Thus far, we have found this technology to be safe, reliable and extremely useful for frameless stereotactic procedures. PMID- 12890977 TI - Stereotactic target localization accuracy in interventional magnetic resonance imaging. AB - OBJECTIVE: To compare stereotactic target determination, based on images obtained from interventional MRI (iMRI), conventional closed MR and CT. METHODS: Stereotactic coordinates for 55 targets in an artificial scull were derived from iMRI scans and compared using CT as the standard. Stereotactic coordinates were also derived from iMRI scans in a series of patients and compared using iMRI fused with CT as the standard. RESULTS: The mean difference between targets in the skull phantom determined from iMRI and CT images was 0.90 +/- 0.28 mm, with a maximum difference of 1.57 mm. The mean difference between targets in the patients derived from iMRI alone and interventional MR fused with CT was 1.39 +/- 0.54 mm, with a maximum difference of 2.47 mm. DISCUSSION: The results indicate that iMRI can be used for stereotactic target localization. PMID- 12890978 TI - Stereotactic interstitial radiosurgery with a miniature X-ray device in the minimally invasive treatment of selected tumors in the thalamus and the basal Ganglia. AB - The aim of this study was to evaluate the role of interstitial radiosurgery (IR) using the photon radiosurgery system (PRS) in the treatment of selected tumors within the thalamus and the basal ganglia. The PRS is a miniature X-ray generator that was developed for interstitial irradiation. This series included 14 patients (5 with glioblastomas, 4 with low-grade astrocytomas and 5 with metastases) harboring spheroidal lesions with dimensions ranging from 13 to 42 mm (mean 30 mm). After stereotactic biopsy, a radiation dose ranging from 6 to 15.4 Gy (mean 11.3 Gy) was delivered at the target volume margins. Follow-up varied from 3 to 26 months (mean 10.2 months). In the group of glioblastomas, 3 patients died (3 12 months after the procedure) because of tumor progression, while the remaining had tumor control. Two patients with metastases died from systemic disease (4-9 months after the treatment), and 3 were alive and well at the end of the study. Local control was achieved in all metastases. Patients with low-grade astrocytomas were well and imaging studies showed tumor control PRS IR is a minimally invasive procedure for the treatment of selected glial or secondary brain tumors. Compared to conventional radiosurgery (brachytherapy and external radiosurgery), PRS IR presents dose delivery characteristics useful for the treatment of tumors in the thalamus and basal ganglia, without inconveniences such as handling radioisotopes, the need of expensive facilities and radiation protection measures. Although the clinical value needs further investigations, PRS IR seems to be effective in metastases while it provides less benefit in malignant gliomas. PRS IR could have a major role in the treatment of low-grade astrocytomas. PMID- 12890979 TI - Long-term efficacy of globus pallidus stimulation for the treatment of Parkinson's disease. AB - OBJECTIVE: To determine the long-term efficacy and safety of globus pallidus internus (GPi) stimulation for Parkinson's disease (PD). BACKGROUND: We previously reported 3-month data for 5 patients who underwent GPi stimulation for PD. We now report long-term data on these 5 patients and 4 additional patients. METHODS: Nine PD patients, 5 men and 4 women, with an average age of 49 years and disease duration of 10 years, underwent GPi stimulation. Six patients had staged bilateral implants and 3 patients had unilateral implants. The mean follow-up was 48.5 months. All patients were evaluated with the Unified Parkinson's Disease Rating Scale (UPDRS) and completed 2-day diaries before and after surgery. RESULTS: There was a 21% improvement in UPDRS Part II (activities of daily living; ADL) scores and a 37% improvement in UPDRS Part III (motor) scores when the longest follow-up in the 'stimulation-on/medication-off' state was compared to the 'medication-off' state at baseline. The UPDRS Part II (ADL) scores improved by 30% and the UPDRS Part III (motor) scores improved by 39% when the longest follow-up in the 'stimulation-on/mediation-on' state was compared to the 'medication-on' state at baseline. As measured by patient diaries, 'on' time increased from 25 to 59% and 'on with dyskinesia' decreased from 42 to 15%. Surgical- and device-related complications included transient hemiparesis in the operating room, postoperative seizures, and implantable pulse generator and lead problems. There were seven device-related events requiring additional surgical procedures. CONCLUSIONS: GPi stimulation continues to be effective for the long term treatment of the disabling symptoms of PD; however, the physician and patient should be aware that device-related problems are not uncommon and additional surgery may be necessary. PMID- 12890980 TI - Bilateral pallidal stimulation for cervical dystonia. An optimal paradigm from our experiences. AB - We sought to identify optimal paradigms of bilateral globus pallidus internus (GPi) stimulation in 3 subsequent patients with severe cervical dystonia. At low frequency stimulation (50-60 Hz) with wide pulse width (500 micros) and high amplitude (4.5-8.0 V), we observed immediate and consistent improvement of dystonia and dystonia-associated pain. Stimulation of the posteroventral portion of the GPi led to pronounced alleviation of dystonia; stimulation of the anterodorsal portion or at the dorsal border of the GPi resulted in significant worsening of symptoms. The therapeutic benefit obtained by using the optimal stimulation parameters continues and has lasted for at least 1 year in each patient. PMID- 12890981 TI - Hardware-related complications after placement of thalamic deep brain stimulator systems. AB - Deep brain stimulation (DBS) has become a common therapeutic approach to patients with movement disorders. We evaluated results from two centers in a large metropolitan area where DBS systems are implanted into the thalamus for patients with tremor. Although all implanted systems led to an improvement in tremor, morbidity related to the hardware system occurred. A total of 23 hardware problems were noted in 66 patients undergoing implantation of 66 DBS systems (27% of patients). The most common problem included breakage of the electrode lead in its extracranial location, system infection, battery or connector problems, lead migration, and hemorrhage. Management was tailored to the specific hardware related problem. Modifications in both surgical technique and hardware design should reduce the incidence of complications. Surgeons who place DBS systems should be aware of the spectrum of problems that can be associated with the device and its placement. PMID- 12890982 TI - Anterior capsulotomy for treatment of refractory obsessive-compulsive disorder: results in a young and an old patient. AB - The objective of this case report was to assess the effect of anterior capsulotomy for obsessive-compulsive disorder (OCD) in 2 patients beyond extremes of age ranges of published radiofrequency capsulotomy. The youngest patient developed OCD at age 10 with increasing symptoms of tension and worry. The symptoms were refractory to medications and behavioral therapy. He underwent anterior capsulotomy at age 18. The older patient was 64 at the time of surgery. His OCD began about age 17 with checking and counting rituals. His obsessions extended into other areas such as fear of injuring people while driving. His work performance was greatly compromised. Despite medication trials his rituals and obsessions intensified. After 47 years of severe symptoms he underwent surgery. The youngest patient returned to high school full-time and graduated. He was able to read and comprehend without obsessing about the meaning of words. His thinking and behavior became symptom free and he married 4.5 years after surgery. His score on the Yale-Brown Obsessive Compulsive Scale (Y-BOCS) after surgery on no medication is zero. The older patient improved gradually without medication. He could play with and touch his grandchildren for the first time. He could drive a car again and his Y-BOCS dropped from 30 preoperatively to 8. Twenty-four months after surgery he is essentially free of obsessions, compulsions and anxiety. We conclude that treatment-refractory OCD may be alleviated by anterior radiofrequency capsulotomy in the young and the old patient. This study expands the documented age range of response from 18 to 64. PMID- 12890983 TI - Linac radiosurgery for brain metastasis of melanoma. AB - PURPOSE: To evaluate the efficacy and toxicity of stereotactic radiotherapy in the treatment of brain metastases of melanoma. PATIENTS AND METHODS: From 1994 to 2001, 25 patients presenting with 61 metastases of cutaneous melanoma were treated with radiosurgery. Median age was 47 years (range: 25-73 years) and median Karnofski performance status 80 (range: 50-100). Twenty patients had one radiosurgery, 5 had two or three. Median metastasis diameter was 21 mm (range: 6 54.4 mm), and median metastasis volume was 1.7 cm(3) (range: 0.4-25.6 cm(3)). Irradiation was delivered by a linear accelerator. Median minimal dose was 14.1 Gy (range: 10-19.4 Gy), and median maximal dose was 20.5 Gy (range: 16-48 Gy). RESULTS: Mean follow-up was 12.6 months (range: 1-85 months). Five metastases progressed (9.8%), 2-12 months after radiosurgery. Three-, 6- and 12-month local control rates were 95 +/- 3, 90 +/- 5 and 84 +/- 7%, respectively. By univariate analysis, only absence of extracranial tumor was a prognostic factor of local control. Three-, 6- and 12-month brain-disease-free survival rates were 75 +/- 9, 68 +/- 11 and 38 +/- 13%, respectively. According to univariate analysis, only the Score Index for Radiosurgery in brain metastases (SIR) was a prognostic factor of brain-event-free survival (p = 0.03). Median survival was 8 months. Three-, 6- and 12-month overall survival rates were 75 +/- 9, 53 +/- 10, and 29 +/- 10%, respectively. According to univariate analysis, extracranial controlled disease status (p = 0.03), and SIR (p = 0.04) were prognostic factors for overall survival. According to multivariate analysis, none was an independent prognosticator for overall survival. Complications were minimal. CONCLUSION: Radiosurgical treatment of brain metastases of melanoma is effective and accurate. The use of radiosurgery alone is an appropriate management strategy for many patients with brain metastases of melanoma. PMID- 12890984 TI - Frameless stereotaxy without rigid pin fixation during awake craniotomies. AB - OBJECTIVE: The accuracy and precision of resection with awake brain mapping is augmented when combined with frameless stereotaxy. Frameless stereotactic assisted surgery, however, typically involves immobilization in Mayfield pins. Rigid fixation, however, may be problematic for individuals undergoing awake craniotomy. We describe an alternate method of frameless stereotaxy without the use of pin fixation. METHODS: The MRI fiducials are placed in proximity to the proposed incision and prepared and draped in the operative field. After craniotomy, the epidural skull clamp (standardly used to support the electrode holder during corticography) is attached. Using the Stealth Navigational System, a spinal reference arc is then clamped onto the skull clamp base. After the fiducials and arc are registered, the probe is used for frameless guidance. RESULTS: In 14 of 15 cases (93%), this technique was successful. One case failed because of close proximity of the fiducials to the incision. The accuracy of the 14 successful cases was less than 4 mm. The error within the lesion itself was always less than 2 mm. Overall, these results are comparable to those achieved using a Mayfield head holder. In all tumor cases, postoperative imaging concurred with intraoperative assessment of a total versus subtotal resection. CONCLUSIONS: By affixing the spinal arc to the skull, the complications and discomfort associated with pin fixation are avoided completely. The patient is free to move without affecting the accuracy. The spine arc, in combination with the skull clamp, provides an efficient, well-tolerated, and accurate method of frameless navigation for the awake patient undergoing craniotomy. PMID- 12890985 TI - Gamma knife radiosurgery for brain metastases: do patients benefit from adjuvant external-beam radiotherapy? An 18-month comparative analysis. AB - OBJECTIVE: To analyze 18 months of results of gamma knife stereotactic radiosurgery in the treatment of brain metastases and determine factors affecting outcome by examining the effectiveness of additional external-beam radiotherapy (XRT). MATERIALS AND METHODS: Between January 2000 and September 2001, 61 patients with 103 tumors diagnosed as cerebral metastases were treated with gamma knife. Mean patient age was 57 years (range = 36-81). Lung carcinoma (55.7%) was the most common primary cancer, followed by melanoma (14.8%) and breast carcinoma (11.5%). Mean KPS of the patients was 70 (range = 50-90). Twenty-seven patients had solitary metastases while 34 had multiple tumors. Forty-three patients (59 tumors in total) received only radiosurgery, while 18 patients (44 tumors in total) had prior XRT. Tumor volume ranged from 0.5 to 33 cm(3) (mean = 9.74 cm(3)). Mean marginal dose prescription to the tumor was 15 Gy (range = 11-21 Gy). RESULTS: Median follow-up was 11 months. Twenty-one patients (34.4%) were alive at last follow-up and 40 (65.6%) had died. Seventeen deaths (42.5%) were reported to be due to progressive brain disease, while 23 deaths (57.5%) were due to uncontrolled primary. Control of the treated lesions was achieved in 45 patients (73.8%) and 84 tumors (81.6%). Mean overall survival of the patients is 8 months (range = 1-19 months). The actuarial 12-month tumor control rate using the Kaplan-Meier method for this series is 68.2 +/- 0.06%. Results of the log rank test revealed that younger age (<55 years), small tumor volume (<10 cm(3)), and increasing tumor dose (>15 Gy) correlated with improved brain disease-free survival (p < 0.05). Overall survival, local tumor control rate and the freedom from brain disease period (based on the appearance of new brain tumors after radiosurgery) were analyzed separately for the groups receiving radiosurgery alone and those with prior XRT to detect any additional benefit of XRT. No statistically significant difference was found between the two groups for any of the considered outcomes. CONCLUSION: Gamma knife stereotactic radiosurgery is a safe and effective treatment option for patients with cerebral metastases. It provides survival benefits and improves quality of life by achieving excellent control of the brain disease, irrespective of patients' age or number of brain tumors. The addition of XRT in younger patients with small brain metastases does not improve survival and/or control of the brain disease. PMID- 12890986 TI - Co-registration of function and anatomy in frameless stereotaxy by contour fitting. AB - We investigated a co-registration algorithm using a contour-fitting procedure to integrate functional data from magnetoencephalography (MEG) and functional magnetic resonance imaging (fMRI) for frameless stereotaxy. In fMRI the shape of the head was reconstructed from anatomical images, in MEG it was scanned using an electromagnetic sensor position indicator. Functional information was transferred to the 3D-MR image set used for frameless stereotaxy by fitting the digitized (MEG) and reconstructed head shape (fMRI) to the 3D-MR images. The mean residual error of the contour fit was 2.3 mm for the MEG and 1.3 mm for the fMRI registration. According to computer simulations, the achievable transformation error is 0.75 and 0.5 mm, respectively. This method enables independent recording of functional and anatomical measurements with a co-registration accuracy better than 2 mm. PMID- 12890987 TI - Neurovascular compression associated with trigeminal neuralgia and systemic arterial hypertension: surgical treatment. AB - OBJECTIVE AND IMPORTANCE: To highlight arterial hypertension as an additional factor favoring surgical indication in neurovascular compression syndromes such as trigeminal neuralgia, hemifacial spasm, and glossopharyngeal neuralgia. CLINICAL PRESENTATION: A 52-year-old woman with trigeminal neuralgia concomitant with systemic arterial hypertension, submitted initially to unsatisfactory conservative treatment, presents tortuousness and enlargement of the vertebral artery (VA) topography on magnetic resonance imaging (MRI). A microsurgical neurovascular decompression of the trigeminal nerve and the medulla was performed, revealing that the compression was due to the superior cerebellar artery instead of the VA. Both the neuralgia and the hypertension were controlled with no need of medication during a 1-year follow-up. CONCLUSION: Surgical indication in neurovascular compression disorders should be reinforced in the presence of concomitant arterial hypertension and a compatible MRI examination. PMID- 12890988 TI - Psychosomatic aspects of pancreatic cancer. PMID- 12890989 TI - Fundamentals of transcription factors and their impact on pancreatic development and cancer. AB - Transcription factors are proteins that regulate gene expression by modulating the synthesis of messenger RNA. Since this process, known as gene transcription, is often the dominant control point in the production of many proteins, transcription factors are key regulators of numerous cellular functions, including secretion, proliferation, differentiation, and apoptosis. Most transcription factors are also the final effectors of signaling pathways that transduce signals from the cell membrane to the nucleus. Therefore alterations in the activity or expression of some transcription factors have a significant impact on the biology of human cells and may lead to the development of diseases. In this article we review this field of research with a particular emphasis on the role of transcription factors in pancreatic development and cancer. PMID- 12890990 TI - Control of pancreatic exocrine secretion via muscarinic receptors: which subtype(s) are involved? A review. AB - The present report gives an overview of the experimental, pharmacological and molecular investigations that have been undertaken during the past two decades to characterize and identify the muscarinic receptor subtype(s) involved in the cholinergic control of pancreatic exocrine secretion in humans and different animal species. The results published in the literature clearly indicate that both M1 and M3 receptors contribute to the regulation of pancreatic enzyme secretion, although contradictory conclusions have been drawn from secretory studies using specific M1 and M3 receptor antagonists in vivo and in vitro. Binding studies using specific M1 and M3 receptor antagonists have supported the existence of both M1 and M3 receptors on pancreatic acinar cells, which was confirmed by the demonstration of specific mRNA for both receptor subtypes in rat pancreatic acinar cells. In addition, experimental evidence exists that nonacinar (possibly presynaptic) M1 receptors also contribute to the control of pancreatic enzyme secretion. The role of the different muscarinic receptor subtypes in the control of pancreatic fluid and bicarbonate output, however, still needs to be clarified. Future research should cover the evaluation of the relative contribution of the different receptor subtypes to the regulation of pancreatic exocrine function, the localization of the receptors involved as well as possible species differences. PMID- 12890991 TI - Effect of FHIT gene replacement on growth, cell cycle and apoptosis in pancreatic cancer cells. AB - The human FHIT gene is altered or lost in many cancers and FHIT has been shown to be a tumor suppressor. However, the mechanism of tumor suppression by the FHIT gene remains unclear. FHIT expression is lost in primary pancreatic cancer and human pancreatic cancer cell lines. To gain insight into the function of FHIT gene, we replaced the FHIT gene in a FHIT-null pancreatic cancer cell line, and established stable fhit-expressing clones. Expression of the exogenous fhit was at similar levels as in other cultured cell lines and fhit protein was found predominantly associated with perinuclear area. fhit replacement resulted in reduced cell proliferation in transfected Panc-1 cells. Cell cycle distribution analysis indicated increased accumulation of G(0)/G(1) phase cells in transfected clones indicating a retardation of cell cycle progression. We observed specific up-regulation of cdc2 and cyclin D3 upon fhit replacement. Furthermore, Bcl-2 family members Bad, Bak, and Bcl-xS protein levels were increased in FHIT transfected clones when compared with Panc-1 cells. Multiplex RT-PCR of apoptosis pathway related genes revealed that Bcl-2 is absent and Bcl- xS message increases in FHIT transfected clones. Our data suggested that exogenous expression of FHIT in Panc-1 cells affects genes regulating cell cycle arrest and apoptosis, and these molecular changes may contribute to the tumor suppressor activity of the FHIT gene. PMID- 12890992 TI - A prospective evaluation of pancreatic exocrine function in patients with acute pancreatitis: correlation with extent of necrosis and pancreatic endocrine insufficiency. AB - AIMS: The aim of this prospective study was to assess pancreatic exocrine function in patients recovering from a first attack of acute pancreatitis, and to evaluate its relationship to severity of attack, extent of pancreatic necrosis and severity of pancreatic endocrine insufficiency. METHODS: Between December 2000 and November 2001, 23 patients were prospectively evaluated. Pancreatic exocrine function was measured by the faecal elastase-1 test and insufficiency was classified as moderately impaired or severely impaired. Pancreatic necrosis was determined by contrast-enhanced CT scan, and its extent was categorised according to Balthazar's classification. The severity of pancreatic endocrine insufficiency was categorised according to insulin dependence. Attacks were classified as mild (n = 16) or severe (n = 7) according to the Atlanta criteria. RESULTS: Pancreatic exocrine insufficiency was significantly more frequent in patients recovering from severe attacks than mild (n = 6, 86% vs. n = 2, 13%; p = 0.002), and in those who developed pancreatic necrosis or pseudocyst than those who did not (6 of 7 patients vs. 2 of 16 patients, and 5 of 5 patients vs. 3 of 18 patients respectively; p = 0.002). The development of exocrine insufficiency correlated strongly with the extent of pancreatic necrosis (r = -0.754, p < 0.001), and the severity of pancreatic endocrine insufficiency (n = 4, r = 0.453, p = 0.03). CONCLUSION: Pancreatic exocrine insufficiency is a common occurrence in patients recovering from severe acute pancreatitis, and its severity correlates with the extent of pancreatic necrosis and the severity of concomitant pancreatic endocrine insufficiency. PMID- 12890993 TI - Predicting fatal outcome in the early phase of severe acute pancreatitis by using novel prognostic models. AB - BACKGROUND/AIMS: Survival in acute pancreatitis and particularly in severe acute and necrotizing pancreatitis is a combination of therapy-associated and patient related factors. There are only few relevant methods for predicting fatal outcome in acute pancreatitis. Scores such as Ranson, Imrie, Blamey, and APACHE II are practical in assessing the severity of the disease, but are not sufficiently validated for predicting fatal outcome among patients with severe acute pancreatitis. The aim of this study was to construct a novel prediction model for predicting fatal outcome in the early phase of severe acute pancreatitis (SAP) and to compare this model with previously reported predictive systems. METHODS: Hospital records of 253 patients with SAP were retrospectively analyzed. 234 patients with adequate data were included to the test set to construct five logistic regression and three artificial neural network (ANN) models. Two models were tested in an independent prospective validation set of 60 consecutive patients with SAP and compared with previously reported predictive systems. RESULTS: The prediction model considered optimal was a logistic model with four variables: age, highest serum creatinine value within 60-72 h from primary admission, need for mechanical ventilation, and chronic health status. In the validation set, the predictive accuracy, determined by the area under the receiver operating characteristic curve value, was 0.862 for the chosen model, 0.847 for the ANN model using eight variables, 0.817 for APACHE II, 0.781 for multiple organ dysfunction score, 0.655 for Ranson, and 0.536 for Imrie scores. CONCLUSIONS: Ranson and Imrie scores are inaccurate indicators of the mortality in SAP. A novel predictive model based on four variables can reach at least the same predictive performance as the APACHE II system with 14 variables. PMID- 12890994 TI - Non-occlusive mesenteric ischemia and its associated intestinal gangrene in acute pancreatitis. AB - BACKGROUND/AIMS: Non-occlusive mesenteric ischemia (NOMI) has been defined as diffuse intestinal ischemia that often results in intestinal gangrene in the presence of a patent arterial trunk. The prevalence and nature of NOMI in acute pancreatitis was investigated. METHODS: A total of 120 consecutive patients with acute pancreatitis managed in the Department of Surgery II, Kumamoto University Medical School, from April 1992 through December 2002, were investigated retrospectively. Among them, 60 patients had the severe form. RESULTS: The overall mortality of acute pancreatitis patients was 8.3% (10/120). The prevalence and mortality of acute pancreatitis associated with NOMI were 6.7% (8/120) and 63% (5/8), respectively, while those of patients with NOMI-associated intestinal gangrene were 4.2% (5/120) and 100% (5/5), respectively. The mortality of patients with severe acute pancreatitis who did not develop NOMI was 10% (5/52). All patients with NOMI-associated intestinal gangrene quickly progressed and subsequently died of multiple organ failure. Plasma creatine phosphokinase (CPK) and lactate levels were elevated significantly in patients with NOMI. CONCLUSION: Acute pancreatitis associated with NOMI was extremely severe. If the plasma CPK and lactate levels are extremely high, NOMI should be suspected. PMID- 12890995 TI - Alterations of systemic endotoxemia over the course of acute edematous pancreatitis. correlation to the advent of an infection? AB - BACKGROUND/AIMS: To define whether bacterial translocation occurs over the course of acute edematous pancreatitis and to correlate its presence to the advent of an infection since data in humans are lacking. METHODS: Thirty-three patients hospitalized over the period January 2000-January 2001 were subjected to venipuncture at regular time intervals for the collection of blood samples for blood culture and for determination of endotoxins and of C-reactive protein. Endotoxins were measured by the Limulus assay and C-reactive protein by nephelometry. RESULTS: A wide range of concentrations of endotoxins was observed over the first 3 days of the disease. Mean (+/-SE) of endotoxins was 4.01 +/- 1.36 and 2.42 +/- 0.95 EU/ml 3 and 6 h, respectively, after admission of afebrile patients. Respective values 3 and 6 h after admission of febrile patients were 3.03 +/- 1.14 and 5.84 +/- 2.28 EU/ml (normal <0.1 EU/ml); these values gradually decreased after the second day. No correlation was found between endotoxins and C reactive protein. Endotoxins were increased as a result of the occurrence of an infection on the third day. CONCLUSIONS: A significant level of endotoxemia is observed over the course of acute edematous pancreatitis, which might be correlated to the advent of the systemic inflammatory response. PMID- 12890996 TI - Nitric oxide regulates bacterial translocation in experimental acute edematous pancreatitis. AB - BACKGROUND/AIMS: The role of nitric oxide (NO) in bacterial translocation (BT) associated with acute pancreatitis is controversial. We investigated the effects of the NO synthase substrate, L-arginine, and the NO synthase inhibitor, N-nitro L-arginine methyl ester (L-NAME), on BT in caerulein-induced acute pancreatitis in rats. METHODS: Acute pancreatitis was induced by subcutaneous injections of caerulein (12 microg/kg) at 6-hour intervals for 2 days. Subcutaneous injections of L-arginine (100 mg/kg) or L-NAME (10 mg/kg) were administeredonce daily for 2 days. At 48 h, pancreatic injury and BT to the mesenteric lymph nodes (MLN), liver, and peritoneum were assessed. RESULTS: Compared with controls, rats that received caerulein injections alone had increased BT to the MLN and pancreatic inflammatory changes. L-Arginine significantly reduced the inflammation and BT caused by caerulein. L-NAME did not significantly alter pancreatic inflammation. Although caerulein + L-NAME-treated rats had increased BT to the peritoneum, MLN, and liver compared with controls, rates of BT did not significantly differ between caerulein alone- and caerulein + L-NAME-treated rats. CONCLUSION: In acute edematous pancreatitis, BT is increased and is regulated by NO. NO substrates limit BT and pancreatic inflammation associated with acute pancreatitis, probably by their bactericidal actions and ability to improve pancreatic blood flow. PMID- 12890997 TI - Vigilin and enzyme expression in isolated pancreatic acini after mellitin and gamma-interferon treatment. AB - BACKGROUND/AIMS: Pancreatitis goes along with changes in exocrine enzyme synthesis and secretion in pancreatic acini. The multi-KH domain protein vigilin is supposed to play an important role in t-RNA trafficking especially in cells with high protein synthesis rates and may reflect the degree of stimulation of translational machinery during pathological processes. In relation to these phenomena we explored in this connection the impact of two different inflammation mediators in a system of isolated rat pancreatic acini. METHODS: Acini were prepared from male Sprague-Dawley rats by collagenase digestion and incubated with mellitin or gamma interferon. Secretion and cytosolic cell content of pancreatic trypsin and amylase as well as the expression of vigilin were determined. RESULTS: The phospholipase A(2) activator mellitin caused morphological alterations and increased release of trypsin and amylase, while vigilin expression and the intracellular content of these enzymes decreased. Gamma-interferon, a cytokine which is involved at different steps in inflammation processes, selectively inhibits the release of trypsin(ogen) while not affecting amylase secretion and vigilin expression. CONCLUSION: Mellitin as well as gamma interferon causes alterations in pancreatic enzyme secretion. Additionally, mellitin seems to influence the expressed gene pattern of pancreatic acini while interferon-gamma has no effect on protein synthesis but enzyme secretion. PMID- 12890999 TI - Differentiation of a pancreatic metastasis of a renal cell carcinoma from a primary pancreatic carcinoma by echo-enhanced power Doppler sonography. AB - In a 70-year-old patient who had been treated for a renal cell carcinoma, a pancreatic mass was detected on CT scan. To differentiate a pancreatic metastasis of the renal cell carcinoma from a pancreatic carcinoma, an echo-enhanced power Doppler sonography was performed. The pancreatic mass demonstrated a strong echo enhancement, proving its hypervascularization. This behaviour favoured the diagnosis of a pancreatic metastasis of the renal cell carcinoma which was confirmed by histology. The principles and the role of echo-enhanced power Doppler sonography in the differential diagnosis between a primary pancreatic carcinoma and a metastasis of a renal carcinoma in the pancreas are discussed. We conclude that this technique can provide an important contribution to the diagnosis in this special instance. However, histology is the standard in the differential diagnosis of pancreatic tumours. PMID- 12890998 TI - Inhibitory effects of human and porcine alpha-amylase on CCK-8-stimulated lipase secretion of isolated rat pancreatic acini. AB - Previously we have demonstrated inhibitory effects of the plant lectin wheat germ agglutinin (WGA) on (125)I-CCK-8 binding to pancreatic AR42J cells as well as on CCK-8-stimulated Ca(2+) release and alpha-amylase secretion of rat pancreatic acini or acinar cells. Therefore, it is entirely conceivable that alpha-amylase having several lectin-like carbohydrate recognition domains can modulate the CCK 8 stimulated lipase secretion. Human alpha-amylase, purified from pancreatic juice by affinity chromatography to homogeneity, and commercial porcine pancreatic alpha-amylase inhibit CCK-8-stimulated lipase secretion of rat pancreatic acini in a concentration-dependent manner. Acarbose, a specific inhibitor of alpha-amylase, was without effect on CCK-8-induced cellular lipase secretion. The data presented here provide evidence for a regulatory function of alpha-amylase in CCK-8-stimulated pancreatic secretion. PMID- 12891000 TI - Primitive neuroectodermal tumor of the pancreas. A case report of an extremely rare tumor. AB - Primitive neuroectodermal tumor (PNET) of the pancreas is extremely rare. Although the diagnosis of PNET is suggested by the light microscopic appearance of the tumor, it should be confirmed by the immunohistochemical evaluation of the c-myc expression and if possible, further determination of the particular chromosome translocation, t(11;22)(q24,q12). In this report, we present a male patient with pancreatic PPNET who had been followed up for 50 months. The related literature is also reviewed. In our case, the pathologic diagnosis was based on the positive immunoreactivity for CD99 in many of the tumor cells. The complementary cytogenetic studies were not possible in the private setting of the patient's treatment. The patient was 31 years old when first operated. Within 4 months of the first operation he had local recurrence. In the third year of his follow-up he had been discovered to have pulmonary metastases and another metastatic tumor in his lung was diagnosed the year after. The metastatic foci were primarily treated by surgical resections. He had chemotherapy after each resection of pulmonary metastatic foci. After 50 months of the initial surgical intervention, he succumbed to widespread thoracic and bone metastases. Because of the extreme rarity of PPNET in the pancreas, and its rather protracted course, we think our case may further contribute to the ever expanding database for this particular entity. PMID- 12891001 TI - Effect of chronic heart rate reduction with ivabradine on carotid and aortic structure and function in normotensive and hypertensive rats. AB - BACKGROUND: A reduction of heart rate (HR) by surgical means or pharmacological agents affects the progression and/or regression of atherosclerotic lesions. Nevertheless, the effect of bradycardia per se on large artery structure and function has never been investigated in rat models of hypertension. METHODS: Four groups of Wistar-Kyoto (WKY) rats and spontaneously hypertensive rats (SHRs) were treated for 28 days either by placebo or by the selective HR-reducing agent ivabradine (8.4 mg/kg/day), a novel compound devoid of inotropic or vasodilating effects and without direct action on the autonomic nervous system. At the end of the follow-up period, intra-arterial blood pressure, carotid pulsatile arterial hemodynamics (echo tracking techniques) and the medial cross-sectional area (MCSA) of the aorta and the carotid artery were determined. RESULTS: In conscious animals, chronic administration of ivabradine significantly reduced HR by 26-30% with no change in tail systolic blood pressure. In anesthetized animals, the decrease in HR and the subsequent increase in the diastolic period were responsible for a decrease in diastolic blood pressure. At the site of the large arteries, ivabradine produced a decrease in the MCSA of the thoracic but not of the abdominal aorta, as well as an increase in pulsatile change of the carotid diameter without change in the isobaric distensibility and MCSA. The changes in pulsatile diameter were significantly larger in WKY rats than in SHRs. CONCLUSION: In normotensive and mainly in SHRs, selective chronic HR reduction by ivabradine is associated with alterations in large arteries involving an aortic antihypertrophic effect. PMID- 12891002 TI - Regulation of matrix metalloproteinase (matrixin) genes in blood vessels: a multi step recruitment model for pathological remodelling. AB - Matrix metalloproteinases (MMPs; matrixins) are a family of structurally related enzymes that collectively promote turnover of all components of the extracellular matrix. Matrix turnover is required for vascular repair, but, if excessive, leads to pathologies that include aneurysm formation and atherosclerotic plaque instability. We review the positive and negative regulation of metalloproteinase gene induction. We propose that multiple steps of gene induction recruit a wider spectrum of MMPs, which may ultimately lead to a transition from matrix turnover to matrix destruction. Studying the detailed mechanisms involved may suggest possibilities for intervening selectively against pathological MMP induction. PMID- 12891003 TI - Vasodilator reactivity to calcitonin gene-related peptide is increased in mesenteric arteries of rats during early pregnancy. AB - The objective of the present study was to determine the effect of early pregnancy on the sensitivity to, and endogenous production of calcitonin gene-related peptide (CGRP). Contractile responses of arteries of 10-day pregnant and nonpregnant rats were studied in myographs. During contractions induced by 40 mmol/l K(+), exogenous CGRP elicited an approximately 30% stronger relaxation in mesenteric arteries in pregnancy, an effect not seen in renal and uterine arteries. Capsaicin treatment during K(+)-induced contractions caused a persistent potentiation of the contractile response in mesenteric arteries, indicating that K(+) stimulates the endogenous release of CGRP. This potentiation was similar in the pregnant and nonpregnant state (+81 +/- 23% and +82 +/- 23%, respectively), suggesting no effect of pregnancy on the endogenous CGRP release. The latter was paralleled by comparable CGRP content in the arteries of both groups, indicating similar tissue CGRP availability. The results of this study support the concept that early pregnancy is associated with a rise in the vascular sensitivity to CGRP in selected areas of the vascular bed without concomitant increase in the vascular CGRP production and release. PMID- 12891004 TI - In vitro study of Starling's hypothesis in a cultured monolayer of bovine aortic endothelial cells. AB - Starling's hypothesis that fluid movement across the microvascular wall is determined by the transmural differences in hydrostatic and osmotic pressures was tested using an in vitro model comprised of bovine aortic endothelial cells grown on a porous support. In all experiments, a 1% bovine serum albumin (BSA) solution was maintained in the abluminal reservoir and the luminal reservoir contained either a 1 or a 5.5% BSA solution. The global osmotic pressure difference across the endothelial layers was thus either 0 or 20.3 cm H2O. When the luminal concentration of BSA was changed from 1 to 5.5% at a hydrostatic pressure differential of 5, 10 or 20 cm H2O, no reverse flow (in the reabsorption direction) was observed even though the hydrostatic pressure differential was far below the global osmotic pressure differential. In another case, the hydrostatic pressure differential was dropped quickly from 20 to 5 cm H2O, while a constant osmotic pressure differential was maintained by 5.5% BSA in the luminal reservoir. A strong transient reabsorption flow was observed over a 30-second period which diminished to undetectable levels within 2.5 min; then a sustained steady-state filtration flow was observed after 20 min. These in vitro experiments support other studies in capillaries showing transient reabsorption that decays to steady-state filtration at longer times. PMID- 12891005 TI - Induction of angiogenesis in omentum with vascular endothelial growth factor: influence on the viability of encapsulated rat pancreatic islets during transplantation. AB - Transplantation of pancreatic islets is proposed as a treatment for type 1 diabetes, but insufficient blood supply can cause the loss of viable grafted islets. In the present study, we investigated the influence of vascular endothelial growth factor (VEGF) on the angiogenesis of omentum during encapsulated islet allotransplantation and consequently on islet survival. Fifty rat islets, cultured for 24 h, were encapsulated in the presence or absence of human VEGF and implanted in the peritoneal cavity of rats (n = 6). After 7, 14 and 28 days of implantation, encapsulation devices with surrounding omentum were removed. Histological analysis of this tissue was performed. Cellular adhesion at the membrane surface was characterized by a phagocytosis test. The morphological aspect of the islets was analyzed and their functionality was evaluated by measuring insulin secretion. At each step of the study, there was a two-fold increase in the number of vessels in the presence of VEGF. In addition, VEGF increased the vessel diameter and the surface area of the angiogenic pedicle. Moreover, the presence of VEGF significantly decreased the distance between the devices and vessels (16.2 +/- 5.6 vs. 51.6 +/- 10.1 microm, p < 0.001). Membrane surface analysis showed a decrease in macrophage adhesion in the presence of VEGF. Furthermore, islet structure and functionality was preserved in the presence of VEGF. Stimulation of angiogenesis of omentum induced by VEGF is associated with preservation of islet viability. Local delivery of VEGF proved to be a relevant approach to ameliorate the outcome of islet transplantation. PMID- 12891006 TI - CD34+ blood cells accelerate vascularization and healing of diabetic mouse skin wounds. AB - Diabetes is characterized by poor circulation and impaired angiogenesis, which appear to contribute to the frequent skin lesions and poor wound healing common in diabetic patients. Therapies to improve circulation commonly improve wound healing in diabetic patients. Administration of circulating CD34+ cells, cells that can function as endothelial cell progenitors, accelerates blood flow restoration to ischemic limbs of diabetic mice. We have investigated the potential of these cells to accelerate revascularization and healing in full thickness skin wounds of hypoinsulinemic (streptozotocin-treated) diabetic mice. Wounds were injected with human CD34+ or CD34- peripheral blood mononuclear cells or no cells, and analyzed for vascularity and healing at various times thereafter. Treatment with CD34+ enriched cells decreased wound size by 4 days after treatment, accelerated epidermal healing, and rapidly and dramatically accelerated revascularization of the wounds compared to controls. Initially increased vascularization was mediated principally by an increase in vessel diameter, but later, both an increase in vascular size and number were observed. These findings indicate that blood-derived progenitors may have therapeutic potential in the treatment of skin lesions in the setting of diabetes, and give insights into how bone marrow cells exert their effects on neovascularization. PMID- 12891008 TI - Study On Metagonimus Yokogawai(Katsurada, 1912) In Korea Vii. Susceptibility Of Various Strains Of Mice To Metagonimus Infection And Effect Of Prednisolone. AB - An experimental study was undertaken to observe the difference in susceptibility of mouse strains to Metagonimus yokogawai infection by estimating it from worm recovery rate and dimension of worms. It was also studied the effects of prednisolone injection on the chronological pattern of worm recovery in ICR mice. The metacercariae were obtained from sweetfish and 300 in each number were given to 5 strains (CBH, A, DBA, C(57)BL and KK) of mice, and after 7 days period, the worms were collected from their intestine. Prednisolone at the dose of 10 mg/kg was injected to ICR mice every other day from 7 days prior to infection until sacrificed at 6 hours to 35th post-infection day. ICR mice infected with M.yokogawai but untreated were used for controls. The success rate in infection of mice ranged 25.0-83.3% by strains, the worm recovery rate 1.2-18.9%, and the average size of worms 0.554-0.683 mm long and 0.214-0.244 mm wide. The higher rates and larger size of worms were observed in KK and C(57)BL strains than others and the difference was statistically significant. In ICR mice for control, the worm recovery rate until 1 day after infection was relatively high (38-66%) but it became much lower (less than 0.7%) during 1-35 days. However, prednisolone injection brought about persistently high recovery rates (16-80%) until 21 days. It was concluded that the susceptibility to M. yokogawai infection is different by strains of mice but it can be elevated by prednisolone injection probably due to suppression of immune responses in ICR mice. PMID- 12891007 TI - Signal transduction in matrix contraction and the migration of vascular smooth muscle cells in three-dimensional matrix. AB - The interaction of vascular smooth muscle cells (SMCs) and extracellular matrix plays important roles in vascular remodeling. We investigated the signaling pathways involved in SMC-induced matrix contraction and SMC migration in three dimensional (3D) collagen matrix. Matrix contraction is inhibited by the disruption of actin filaments but not microtubules. Therefore, we investigated the roles of signaling pathways related to actin filaments in matrix contraction. SMC-induced matrix contraction was markedly blocked (-80%) by inhibiting the Rho p160ROCK pathway and myosin light chain kinase, and was decreased to a lesser extent (30-40%) by a negative mutant of Rac and inhibitors of phosphatidylinositol 3-kinase (PI 3-kinase) or p38 mitogen-activated protein kinase (MAPK), but it was not affected by the inhibition of Ras and Cdc42-Wiskott Aldrich syndrome protein (WASP) pathways. Inhibition of extracellular-signal regulated kinase (ERK) decreased SMC-induced matrix contraction by only 15%. The migration speed and persistence of SMCs in the 3D matrix were decreased by the inhibition of p160ROCK, PI 3-kinase, p38 MAPK or WASP to different extents, and p160ROCK inhibitor had the strongest inhibitory effect. Our results suggest that the SMC-induced matrix contraction and the migration of SMCs in 3D matrix share some signaling pathways leading to force generation at cell-matrix adhesions and that various signaling pathways have different relative importance in the regulations of these processes in SMCs. PMID- 12891009 TI - [Epidemiological studies on Metagonimus yokogawai infection in Samcheok-gun, Kangwon-do, Korea] AB - An epidemiological study on Metagonimus yokogawai infection was performed in Samcheok-gun, Kangwon province during the period from November 1983 to October 1984. Formalin-ether concentration technique and Stoll's dilution egg counting method were employed to figure out the prevalence and infection intensity of M. yokogawai infection. The results are summarized as follows: The prevalence of M. yokogawai infection was 13.3 % among 1,172 examined; 18.0 % (114 out of 632) in male and 7.8 % (42 out of 540) in female. The prevalence rates were different by stream; 15.5 % in Ohsip-chon (stream), 11.8 % in Maeup-chon and 1.6 percent in Gagok-chon. Along the basin of Oship-chon; 8.2 % at the lower reaches in Samcheok eup, 25.9 % at the middle reaches in Miro-myon and 6.2 % at the upper reaches in Dogeoe-eup, respectively. By age group, the highest rate (18.3~20.2 %) was observed in 30~49 years of age (male : 24.1~28.7 %, female 11.2~13.2 %). By social strata, the positive rate was 15.0 % (120 out of 799) in general inhabitants and 9.7 % (36 out of 373) in school children. Intensity by EPG count in feces was 419 in average; the light infection (less than 400 EPG) was 70.4 %, moderate infection (401~1,000 EPG) 19.4 % and heavy infection (more than 1,001 EPG) 10.2 percent respectively. All the 67 sweetfish (length 10.3~18.5 cm) which were caught from Ohsip-stream were infected with metacercariae of M. yokogawai and the number of metacercariae detected were 382 per fish in average (323 in June and 382 in September). Experiences of intake of raw fresh-water fish [Plecoglossus altivelis and Leuciscus (Tribolodon) sp.] were present in 50.3 % (354 out of 704) of the inhabitants along Ohsip-stream according to questionnaire study. The results of this survey indicated that the middle reaches of Ohsip stream in Miro-myon, Samcheok-gun is the endemic region for M. yokogawai infection. PMID- 12891010 TI - Comparative Morphology Of Eggs Of Heterophyids And Clonorchis Sinensis Causing Human Infections In Korea. AB - In order to provide some clues for differential diagnosis of trematode infections in fecal examination, the comparative morphology of eggs of 5 kinds of heterophyid flukes (Metagonimus yokogawai, Heterophyes heterophyes nocens, Heterophyopsis continua, Stellantchasmus falcatus and Pygidiopsis summa) and Clonorchis sinensis was studied. The eggs were obtained from distal portion of uteri of worms which were recovered from men after treatment. The characteristic shape and apperance of each kind of eggs were observed in detail under light microscope, and their length and width measured and compared one another. The results are as follows: Eggs of C. sinensis are elongated ovoidal in shape with attenuated anterior end, 25.3-33.2 (28.3 in average) micrometer long and 14.2 17.4(15.9) micrometer wide with length /width ratio of 1.60-2.00 (1.78). They differ from all heterophyid eggs in that they have prominent wrinkling (muskmelon pattern) at their shell surface. P. summa eggs are ovoid to pyriform in shape and characterized by the smallest size of all kinds examined, 19.8-22.9(21.6) micrometer long and 11.1-13.4 (12.1) micrometer wide and the ratio 1.63-1.99 (1.78). Eggs of S. falcatus are elongated ovoidal and most slender form, 25.3 29.2 (27.2) micrometer long and 11.1-13.4 (12.5) micrometer wide with the ratio of 2.00-2.57 (2.17). Eggs of M. yokogawai are ellipsoid to elliptical in shape with round both ends, 26.9-31.6 (28.5) micrometer long and 14.2-18.2 (16.8) micrometer wide with the ratio of 1.48-2.11(1.70). H. continua eggs are oval in shape, sometimes similar to M. yokogawai or H. h. nocens eggs, however, the relative breadth is broadest among all kinds, with maximum width at posterior half portion. They are 23.7-27.7 (25.0) micrometer long, 15.8-18.9 (16.4) micrometer wide with the ratio of 1.33-1.75 (1.53). Eggs of H. h. nocens are ellipsoid to ovoid in shape but sometimes more slender than M. yokogawai and have slightly pointed both ends. They are 23.7-29.2 (25.7) micrometer long, 14.2-15.8 (15.4) micrometer wide, and the ratio 1.50-2.06 (1.67). From the results, it is concluded that eggs of 5 kinds of heterophyids and C. sinensis can be morphologically differentiated one another, however, careful observation and measurement on sufficient number of eggs are needed. PMID- 12891011 TI - [Studies On Intestinal Trematodes In Korea Xiv. Infection Status Of Loaches With Metacercariae Of Echinostoma Cinetorchis And Their Development In Albino Rats] AB - The metacercariae of Echinostoma cinetorchis (Trematoda: Echinostomatidae) were found infected in the loaches (Misgurnus anguillicaudatus) purchased from two local markets in Korea. Identification of the metacercariae was done after obtaining adult worms from experimentally infected albino rats. Brief course of worm development in rats was also observed up to 28th post-infection day. The results are as follows: The metacercariae of E. cinetorchis were detected from 5 (10.9 %) out of 46 loaches examined and the average metacercarial burden per infected loach was 34.4. They were always found from the posterior abdominal wall. The worm recovery rate from 8 rats was 24.6 % in average. It was observed that the rate decreased according to age of infection; from 40.7 % after 6 days to 15.0 % after 28 days. The adult worms were morphologically characterized by the presence of head crown with dorsally uninterrupted collar spines, 37-38 in number, and the tendency of migration or disappearance of testes. So that they were identified as Echinostoma cinetorchis Ando et Ozaki, 1923. The development of worm was very rapid during 6-10 days after infection and became much slower thereafter. During the rapid worm growth the development of genital organs was more prominent than that of non-genital organs. From the results it is concluded that the loach is involved in the life cycle of E. cinetorchis in this country and may take a role for infection source to animals and man. It is also confirmed that migration or loss of testes is the distinct specific character of this echinostomatid fluke. PMID- 12891012 TI - [Survey On Encysted Cercaria Of Trematodes From Fresh-Water Fishes In Tongjin Riverside Areas In Korea] AB - In an attempt to clarify the epidemiological feature of distomiasis in Tongjin riverside area, the prevalence of distomiasis in the residents and infection rates of the metacercariae in fresh-water fishes were investigated at the upper, middle and lower reaches of the river from January to April 1984. The results obtained were summarized as follows: Out of a total of 931 fresh-water fishes which composed of 33 different species, 611 fishes (65.6 %) of 31 species were found positive with digenetic trematode metacercariae of 16 different species, and there were some differences in infection rates of the metacercariae among the fishes in the 3 parts of the river; 53.8 percent in upper, 80.7 % in middle, and 61.0 % in lower reaches, respectively. Infection rates of the metacercariae of Exorchis oviformis, Metagonimus yokogawai, Echinochasmus japonicus, Metorchis orientalis and Clonorchis sinensis in the fishes were 48 %, 29 %, 11 %, 7.9 % and 6.3 %, respectively. The average number of the encysted larvae of Clonorchis found in fish body/gram showed 4.44 in Pseudorasbora parva, Gnathopogon coreanus (1.2), Microphysogobio yaluensis(0.76), Abbottina springeri(0.4), Acanthorhodeus asmussi (0.21) and Cultriculus eigenmanni (0.17), respectively. The average number of the metacercariae of Metagonimus found in fish body/gram disclosed 34.01 in Zacco platypus, Zacco temmincki (16.46), Carassius carassius (5.35), Moroco oxycephalus (1.54), Aphyocypris chinensis (1.5) and etc., respectively. Detection rates of the eggs of Clonorchis and Metagonimus among residents were 1.1 % and 0.8 %, respectively, out of a total 923 persons. PMID- 12891013 TI - [Immunological Responses By Soluble Egg Antigen Of Schistosoma Mansoni In Mice] AB - This experiment shows cellular and humoral immune responses induced by soluble egg antigen of Schistosoma mansoni, that is, change of the number of peripheral blood eosinophil, delayed hypersensitivity measured by the degree of ear swelling, granulomatous change of liver tissue and elevation of serum antibody titer by ELISA. SEA was given continuously by the insertion of a mini-pump into peritoneal cavity of mouse. In control group, same pump with HGG was inserted. New pump was exchanged once in two weeks and followed the result until 9 weeks after mini-pump insertion. 1. Highest peripheral blood eosinophil level was recorded at 2-3 weeks after SEA pump insertion. 2. Maximum ear swelling was observed at 2 weeks and then decreased gradually. 3. In liver tissue, several granulomas without egg were formed at 4 weeks. 4. Serum antibody titer was elevated from 4 weeks after SEA pump insertion. PMID- 12891014 TI - [Prevalence Of Intestinal Helminthic Infections And Skin Tests For Paragonimus And Clonorchis In Tuberculosis Patients] AB - This study was conducted to evaluate prevalence of intestinal helminthic infections and skin tests and Paragonimus and Clonorchis in tuberculosis(TB) patients in Korea. Stool examinations by formalin-ether technique and intradermal tests were carried out in 2 areas; Masan TB hospital and Kong-Ju TB hospital. 1. The positive rate of intestinal helminths of any kind was 51.7 % among 329 patients in Masan hospital, but 18.1 % among 215 patients in Kong-Ju hospital. 2. The infection rates of Trichocephalus trichiurus and Clonorchis sinensis were 20.7 % and 17.6 % respectively in Masan hospital patients, but the infection rates of T. trichiurus and C. sinensis were 6.5 % and 6.0 % respectively in Kong Ju hospital patients. 3. In skin tests, positive reactions to Paragonimus and Clonorchis antigens were 22.0 % and 37.6 % respectively in Masan hospital patients. On the contrary, Kong-Ju hospital patients showed the positive rates of 15.2 % for Paragonimus and 27.2 % for Clonorchis respectively. PMID- 12891015 TI - A Study On The Parasitic Helminths Of Domestic Duck(Anas Platyrhynchos Var. Domestica Linnaeus) In Korea. AB - During February 1982 to Septemner 1983, a total of 105 domestic ducks (Anas platyrhynchos var. domestica Linnaeus) were collected from Ichon Gun, Puasn City, Chunchon City, Yanggu Gun, Taejeon City and unknown areas in Korea. In the results, six trematode and one nematode species of helminth parasites were obtained and described as follows: Amphimerus anatis Yamaguti, 1933, Echinostoma miyagawai Ishii, 1932, Echinochasmus japonicus Tanabe, 1926, Cryptocotyle sp., Notocotylus attenuatus (Rudolphi, 1809) Kossack, 1911, Apatemon sp, and nematode species, Heterakis gallinarum Schrank, 1788. From this collection, Amphimerus anatis Yamaguti, 1933, Echinochasmus japonicus Tanabe, 1926, Genus Cryptocotyle Luhe, 1899 and genus Apatemon Szidat, 1928 were firtstly recorded in Korea. PMID- 12891016 TI - Serological Diagnosis Of Human Sparganosis By Means Of Micro-ELISA. AB - Seven cases of surgically proven sparganosis were serologically tested by means of micro ELISA for their specific IgG antibody levels. For that purpose, crude saline extract of spargana from snake, Natrix tigrina lateralis was prepared and used as antigen. The sparganosis sera were also tested with Paragonimus and Cysticercus antigens to observe the cross reactivity. A total of 71 sera from normal control, ectopic and pulmonary paragonimiasis, clonorchiasis, cysticercosis and Taenia saginata cases were also included. Except for one case of old calcified infection, all of 6 human sparganosis showed higher serum levels of specific IgG antibody when the differential point of positive reaction was set at the absorbance value of 0.25 (the sensitivity being 85.7%). In control and other helminthic infections, all except 3 cases of T. saginata infection showed negative reaction to sparganum antigen (the specificity being 95.7%). None of sparganosis cases showed cross reactivity to Paragonimus and Cysticercus antigens. Undiluted cerebrospinal fluid also showed high levels of antibody when central nervous system was invaded. The serologic diagnosis by means of micro ELISA could be a useful tool in epidemiological study of human sparganosis in susceptible population, as well as in individual diagnosis. PMID- 12891017 TI - [Migration And Distribution Of Spargana In Body Of Experimentally Infected Mice] AB - The migration and distribution pattern of spargana in mouse body was observed after experimental infection through mouth. The spargana were obtained from the snake, Natrix tigrina lateralis, caught in Hoengseong-gun, Kangwon-do. A total of 28 male mice (ICR strain), 21~25g in body weight, were fed each with 5 scolices (and necks) of spargana and killed after 10 minutes to 14 days. Systemic autopsy was performed on each mouse to recover the spargana. The results are as follows: The spargana were found to penetrate into the stomach or duodenal wall of mice as early as 10 minutes after infection. They completed the penetration within 30 minutes and appeared in abdominal cavity. It was observed that spargana did not migrate tangentially along the gut wall but directly perforated the wall. After 1 hour to 1 day the majority of spargana distributed in abdominal cavity of mice except few which migrated to muscles or subcutaneous tissues. It was within 7 days that nearly all of the spargana migrated to subcutaneous tissues. Out of total 28 in number found from subcutaneous tissues, 13 distributed around neck region, 12 around trunk and other 3 on head of mice and the most common sites were submandibular and subscapular areas. There was nearly no host tissue reaction to migrating spargana. The initial length of spargana given was 4 mm in average but it increased to 12 mm after 7 days and to 35 mm after 14 days. The results suggest that spargana orally given to mice penetrate the gut wall within 30 minutes followed by escaping into abdominal cavity, and after passing through thoracic cavity or abdominal wall they finally localize in subcutaneous tissues chiefly around neck region within 7 days. PMID- 12891018 TI - Distribution of cestodes in the digestive tract of Indian Hill-stream fishes. AB - The distribution of Bothriocephalus sp., Guptaia sp., Mackiewiczia sp., Polyonchobothrium sp., Ptychobothrium sp., and Senga sp. in the alimentary tract of nine Indian hill-stream fishes are described. Though the region around pyloric sphincter was preferred by most cestodes, Senga sp. enabled its existence even in the latter part of intestine apparently because of its well developed adhesive apparatus on scolex. PMID- 12891019 TI - Nematode fauna of high altitude avian hosts in Garhwal Himalayan Ecosystems I. Eustrongylides spinispiculum n. sp. and revised key to the species of genus Eustrongylides Jagerskiold (1909). AB - Analysis of variance has been applied as a new tool for precise substantiation of taxometric differences between Eustrongylides spinispiculum n. sp. and close species, as indicated by the polythetic divisive classificatory system. A revised key to the species of genus Eustrongylides Jagerskiod (1909) has been presented. PMID- 12891020 TI - A human case infected by the larva of Terranova type A in Korea. AB - A human case infected with Terranova type A larva was found in Korea. The patient was a 23-year old soldier of the Korean Army and the chief complaint was acute abdominal pain. The pain was chiefly at right lower quadrant. Appendectomy was performed under the clinical impression of acute appendicitis. However, during the surgery, a nematode larva was found moving on the serosal surface of terminal ileum. The worm was 25.76mm long and 0.66mm wide, and had the intestinal cecum reaching to anterior one-third level of ventriculus and a mucron at posterior end. Therefore, it was diagnosed as Terranova type A larva. This is the first human case of Terranova type A larva infection in Korea. PMID- 12891021 TI - [Experimental meningoencephalitis by Naegleria fowleri in mice] AB - Experimentally, primary amoebic meningoencephalitis (PAM) is induced by Naegleria fowleri in mouse and development of PAM may be influenced by the strain, weight and sex of mouse, and inoculum size of N. fowleri trophozoite. In this paper, the effect of these factors on PAM development of mouse was studied. N. fowleri trophozoites, strain 0359, were introduced into mouse intranasally under secobarbital anesthesia (0.05 mg/g). PAM was developed more frequently in BALB/c mouse than ICR mouse. The survival time of mouse with PAM was influenced by the weight, that is, it was shorter in 15 g mouse than in the heavier groups. No difference was observed on PAM development according to sex. In case of inoculated amoeba, PAM incidence of 0.5 x 10(4) was markedly decreased. PMID- 12891022 TI - [Ultrastructural observation of Naegleria fowleri trophozoite in mouse brain and axenic culture] AB - Present study was undertaken to elucidate the changes of the ultrastructure of Naegleria fowleri trophozoite in brain tissue of mice and culture medium. Naegleria fowleri, 0359 strain, which used in this study was cultured in axenic liquid medium, CGVS medium. Each mouse was inoculated with 1 x 10(5) amoebas intranasally under secobarbital anesthesia, and sacrificed on 7th day after the infection. Comparative observation of the ultrastructure of the amoebas in axenic culture and experimentally infected mice brain was done with transmission electron microscope. The results are summarized as follows: 1. The amoebas in mouse brain tissue were round in outline, whereas those of amoebas from axenic culture showed irregular appearance. 2. Mitochondria in the amoebas from axenic culture was oval, round and cylindrical shape and darkly stained, whereas those of the amoebas from mouse brain tissue showed dumbbell shape together with above forms. The stain was not unique, but light and/or dark. 3. Rough endoplasmic reticulum of amoebas in brain tissue was tubular, but from culture it was vesicular or tubular in shape. 4. Empty vacuoles were demonstrated in amoebas from culture, while food vacuoles with myelinated structures were abundant in those from tissue, suggesting a strong phagocytic activity. 5. Mouse brain tissue infected were extensively destroyed, and polymorphonuclear leukocytes were infiltrated predominantly with inflammatory lesion. Amoebas were observed in the vicinity of the capillary. PMID- 12891023 TI - An outbreak of malaria in penguins at the Farm-land Zoo. AB - In June of 1983 authors had investigated the occurrence of the avian malaria in the Humboldt penguins imported from Japan to the Farm-land Zoo in central district of Korea. The disease was characterized by acute course and high mortality. The peripheral blood smears from the affected penguins demonstrated different developmental stages of Plasmodium sp. in the mature erythrocytes. The predominant gross lesions noticed were pulmonary and epicardial edema and hepato splenomegaly. Microscopically the lesions were characterized by extensive reticuloendothelial cell hyperplasia with striking feature of exoerythrocytic schizogony affecting a variety of tissues. The report also signifies the first description of a disastrous epizootic of avian malaria in Korea. PMID- 12891024 TI - [Head Louse Infestation Among The Students In Yongyang-Gun, Kyongsangbuk-To] AB - The purpose of this study is to know the present situation of head louse infestation among the students in Yongyang-gun, Kyongsangbuk-to From May 11 to June 20, 1984, a total of 5,937 students in 23 primary and 3 middle schools were examined to identify the adult worms of head louse and/or their nits on the hair. The results are as follows: 1. The overall infestation rate was 44.5 percent and the rates ranged from 19.6 percent to 88.6 percent by each school. 2. In primary schools, the rate was 53.0 percent and in middle schools, 27.1 percent. 3. The school boys' rate was 33.2 percent and the school girls' 55.6 percent, and the difference was statistically significant (p<0.001). 4. The rates were different by grades in middle schools. 5. The rates were different by areas (Myons) (p<0.001). 6. The adult worms collected from the students were Pediculus humanus var. capitis. PMID- 12891025 TI - [Study on Metagonimus yokogawai(Katsurada, 1912) in Korea VII. Electron microscopic observation on the tegumental structure] AB - This study was performed to observe the chronological changes in the tegumental structure of M. yokogawai using scanning electron microscope. The subjected worms were excysted metacercariae obtained from sweetfish, and 2-day, 1-week and 4-week old worms experimentally reared in albino rats. The results are as follows: The tegument of excysted metacercariae showed many transverse shallow rugae, which were more remarkable in posterior half body, i.e., posterior to ventral sucker. The whole surface was armed with many scale-like spines; 7~8 pointed ones on anterior body and 2~3 pointed on posterior body. The ciliated knob-like papillae (Type I) were abundant around oral and ventral suckers, which grouped 2, 3 or 4 in number in most cases. A few round swellings of tegument (Type II) were observed only on oral sucker. The tegumental surface of 2-day old worms showed deeper rugae, and the anterior half covered with knob-like processes of distal cytoplasm and the posterior half with cobblestone-like ones. Interspinous space became more wide and 9 pointed spines appeared on anterior dorsal surface. The sensory papillae enlarged but not changed in their distribution. The tegument of 1-week old worms revealed knob-like cytoplasmic processes in posterior half body and velvety ones around oral sucker. The scale-Iike spines of anterior half body changed remarkably to the slender ones of posterior body at the level of ventral sucker. In dorsal surface, the arrangement of the Type I papillae were bilaterally symmetrical. The tegument of 4-week old worms were finely differentiated and the posterior tegument covered with velvety cytoplasmic processes. The spines had remarkably grown in length and width but the density remained nearly unchanged. The papillae also became larger but their morphology and distribution were not different from younger worms. However, the round elevation of cytoplasmic ridges (Type III papilla) appeared bilaterally on inner wall of oral sucker, approximately 8 in number. From the above results, it is considered that the tegument of juvenile M. yokogawai continued to differentiate until 4 weeks after infection. PMID- 12891026 TI - [Study on sensory papillae of Metagonimus yokogawai cercaria] AB - A number of studies on the papillae of cercariae of trematodes reported that the papillar patterns (or chaetotaxy) of cercariae might be an excellent method to attain better understanding of the digenetic trematodes (Richard, 1971; Short and Cartrett, 1973; Bayssade-Dufour, 1979). The present study was aimed to determine the number, distribution pattern and structure of the sensory papillae of Metagonimus yokogawai cercariae, and to elucidate the chaetotaxy of this digenetic trematode. M. yokogawai cercariae were pipetted from a vial in which infected snails (Semisulcospira libertica) had been kept for 3 hours. The snails were collected from an endemic area of M. yokogawai, Boseong river in west southern part of Korea. Observations of papillae were based on light microscopy of those stained with silver nitrate, and on scanning electron microscopy. The results are summarized as follows: All papillae observed were uniciliated. Cilia in anterior tip were shorter than the others in other portions. The body papillae were arranged in essentially symmetrical patterns. Total number of the papillae was 126(63 pairs) in average; anterior tip 40(20 pairs), ventral 20(10 pairs), lateral 42(21 pairs), and caudal 8(4 pairs). The chaetotaxy of M. yokogawai cercaria was; C(I) cycle (3+3C(I)V, 2+2C(I)L, 2+3C(I)D), C(II) cycle(2C(II)V, 1C(II)L, 2C(II)D), C(III) cycle (1+lC(III)V, 1C(III)L), C(IV) cycle (1C(IV)V, 1C(IV)L) in cephalic region: A(I)(lA(I)V, l+2A(I)L, 1A(I)D), A(II)(1A(II)V, 1+3A(II)L, lA(II)D), A(III) {lA(III)V, l+lA(III)L, lA(III)D) and A(IV)(lA(IV)V, 2A(IV)L) in antacetabular region: 1M(I)V and 2M(I)L in median: 1+lP(I)L, 1P(II)L, lP(II)D lP(III)L, lP(IV)L and 1P(IV)D in postacetabular region: 2-2-2-2 in caudal region. PMID- 12891027 TI - [Studies on intestinal trematodes in Korea X. Scanning electron microscopic observation on the tegument of Fibricola seoulensis] AB - A scanning electron microscopic study was performed to observe the tegumental surface of adult Fibricola seoulensis. The adult worms were collected from the small intestine of mice 5 days to 3 weeks after experimental infection with the metacercariae. The metacercariae were obtained from the viscera of the snakes, Natrix tigrina lateralis, by artificial digestion technique. The results were as follows: The tegument of anterior body was covered with cobblestone-like cytoplasmic processes and that of posterior body showed finger-like processes. The posterior body had 4-5 large transverse wrinklings which formed many discontinued shallow rugae. The entire surface of anterior body was regularly arranged with the spines of which tips diverged into 3 to 4 points. They were densely packed in anterior mid-median portion of dorsal surface where appeared a few spines indented upto 5 points. Farther laterally and posteriorly from this portion, the pointed spines were more sparse and became single tipped and extended to anterior one-third of posterior body. The posterior surface of oral sucker was armed with 50-60 spines having 2-3 tips and ventral sucker also covered with such spines. On anteriormost dorsal surface arranged 60-70 spade shaped spines. The tribocytic organ was armed with many stout recurved pile-like spines arranged radially. There were 3 types of sensory papillae. The ciliated knob-like (Type I) papillae were almost bilaterally symmetrical in ventral and dorsal surfaces of anterior body, and abundant especially around bases of oral and ventral suckers, tribocytic organ, and in lateral margins of anterior body. About 24 non-ciliated round swellings (Type II) were observed around each lip of oral and ventral suckers. The plate-1ike elevated papilla without cilium (Type III) was found to distribute only in posterior body. These 3 types of papillae seem to be tangoreceptive and/or rheoreceptive in function when their morphology and distributions are considered. PMID- 12891028 TI - [A study on the fine structure of Clonorchus sinensis, a liver Fluke V. The mature spermatozoa] AB - An ultrastructural study on the mature spermatozoa of Clonorchis sinensis was carried out. For this study, the liver flukes were collected from the livers of rabbits and rats artificially infected with the metacercariae obtained from the fresh water fish, Pseudorasbora parva. Six-month old worms were used. The collected liver flukes were washed with 0.85 percent saline solution and then immediately moved to cold 2 percent glutaraldehyde buffered with 0.l M Millonig's phosphate buffer (pH 7.4). The materials were dissected into appropriate pieces in the fixative about 30 minutes after beginning of the fixation. Two hours later the materials containing the seminal receptacle were rinsed several times with the buffer and were secondarily fixed with cold, buffered 1 percent osmium tetroxide for 2 hours. The fully fixed tissue blocks were dehydrated in a series of graded concentrations of acetone and were embedded in Epon 812 mixture. Thin sections obtained from LKB-5 ultramicrotome were stained with uranyl acetate and Reynold's lead citrate. Observations of the sections were carried out with JEM 100CX II electron microscope. In general, the mature sperm was long thread-like form with a sickle-shaped head. According to the longitudinal sectioned view of the sperm tail, the nucleus seemed to be spirally coiled and run a little far along the tail. The acrosome was not observed. The cytoplasm of the tail was biflagellated as usual in trematodes. Unlike other platyhelminth spermatozoa, the sperm tail of Clonorchis sinensis showed the [9+2] pattern in the microtubular arrangement. The mitochondria with poorly developed cristae were observed throughout the middle piece. The middle piece of the tail showed dull ladder or triangular shapes with the two flagella at the bottom. But, the principal piece of the tail was slightly flattened cylindrical shape with two flagella within the cytoplasm. The end piece was uniflagellated. It was not clearly identified whether the end piece was subdivided into two by flagellum or the lengths of the two flagella were different. The glycogen granules were rich in the cytoplasm throughout the length of the spermatozoa. These granules might be the energy source for the movement of the spermatozoa. PMID- 12891029 TI - Studies on intestinal trematodes in Korea XI. Two cases of human infection by Heterophyes heterophyes nocens. AB - Two human cases of Heterophyes heterophyes nocens infection were proved by identifying adult worms after treatment with bithionol or praziquantel in 1983 in Korea. They are 37 (Case 1) and 24-year old (Case 2) males whose residence or native village is a southern coastal area in Kohung-gun, Chollanam-do. The Case 1 had the gastrointestinal symptoms such as epigastric pain and indigestion, and the Case 2 heart problems such as arrhythmia and ventricular premature beat in EKG and digestive symptoms such as diarrhea and abdominal pain. After the treatments, 35 and 67 specimens of H. heterophyes nocens respectively were collected from the diarrheal stools through stereomicroscopy. The Case 1 was concommitantly infected with Clonorchis sinensis and the case 2 with 3 other kinds of heterophyid flukes and with Diphyllobothrium latum. The cases said to have eaten raw brackish water fish such as mullets (Mugil cephalus), perches (Lateolabrax japonicus) and gobies (Acanthogobius flavimanus) which are the intermediate hosts of H. heterophyes nocens in Korea. PMID- 12891030 TI - Studies on intestinal trematodes in Korea [XII. Two cases of human infection by Stellantchasmus falcatus. AB - Two cases of human infection by Stellantchasmus falcatus (Heterophyidae) were identified by collection of adult worms during the treatment of some tapeworm infections in Korea. The cases were 24 (Case 1) and 55-year old (Case 2) males residing in Seoul. The Case 1 had gastrointestinal troubles such as diarrhea and abdominal pain, and heart problems such as palpitation and arrhythmia, revealing the eggs of Diphyllobothrium latum in the feces. The Case 2 complained indigestion and discharge of tapeworm (Taenia saginata) segments. Praziquantel at the dose of 15-20 mg/kg body weight and 30 g magnesium salt were given to them for treatment of the tapeworm infection and the discharged strobilae were identified. However, concommitantly with the tapeworms, 188 and 5 specimens of S. falcatus were collected from Case 1 and Case 2 respectively through stereomicroscopy of the diarrheal stools. The Case 1 was infected also with 3 other kinds of heterophyid flukes. They said to have eaten raw brackish water fish such as mullets which are considered to be the source of heterophyid fluke infections. PMID- 12891031 TI - Studies on intestinal trematodes in Korea XIII. Two cases of natural human infection by Heterophyopsis continua and the status of metacercarial infection in brackish water fishes. AB - Two cases of natural human infection by Heterophyopsis continua (Heterophyidae) were identified by collection of adult worms in 1983 in Korea. And in order to know the source of infection a study on fish intermediate hosts was performed along the southwestern coastal areas. The cases were 24 and 50-year old males residing in southern coastal areas. They had the clinical complaints of non specific gastrointestinal symptoms such as epigastric pain and one of them also complained the discharge of tapeworm segments. Praziquantel in single dose of 10~15 mg/kg was given followed by purgation with mangnesium salt and 2 and 46 specimens of H. continua were collected from the diarrheal stools. They said to have eaten raw flesh of several kinds of brackish water fishes. The results of fish examination for metacercarial infection were as follows: Seventeen (63.0%) out of 27 perches (Lateolabrax japonicus) and 10 (29.4%) out of 34 gobies (Acanthogobius flavimanus) harboured the metacercariae but none of 42 mullets (Mugil cephalus) were found to harbour them. The average metacercarial density in perches was 55.4 and 18.7 per fish respectively. And the majority of the metacercariae were collected from the muscle of body portions in these fishes. After the metacercariae were experimentally fed to two rats and one young dog, two adult worms were recovered from the dog while none from the rats. These worms were also indentified as H. continua. From the results it was concluded that the brackish water fishes which the human cases said to have eaten were the source of infection. PMID- 12891032 TI - Fifteen human cases of Fibricola seoulensis infection in Korea. AB - A mass fecal examination was performed in Seoul, Korea from December 1983 to March 1984 and 15 egg positive cases of Fibricola seoulensis were discovered. They were treated with praziquantel(Distocide(R)) 20 mg/kg single dose and magnesium salt for purgation. The adult worms of F. seoulensis were collected from 6 of them. The number of the flukes ranged 1-78 by the case. The cases had no clinical mainfestation at all. They were all Korean males in twenties, and had the histories of eating the roast or raw snakes or frogs. Fibricoliasis is expected to be prevalent among the snake-eaters in Korea. Further cases will be detected by sedulous differentiation of thin-shelled immature operculated eggs of 0.081-0.102mmn length. Praziquantel seems to be an excellent chemotherapeutic for fibricoliasis. PMID- 12891033 TI - Two cases of human infection by adult of Spirometra erinacei. AB - Two cases of human infection by adult Spirometra erinacei were proved by collection of worms in Korea. The patients were 24 and 21-year old males residing in a mountainous village in Kangwon-do. They had the clinical complaints of abdominal pain and epigastric discomfort, revealing the diphyllobothriid eggs, 53~59 x 37~42 micrometer in size, in their stools. They were treated with 500 mg atabrine and purgated with magnesium salts, and 3~4 hours later several chains of tapeworm segments were recovered from the diarrheal stools. The recovered worms revealed the morphological characteristics of spiral-form or coiled uteri, separated vaginal opening from the cirrus sac, incorporated seminal vesicle into the cirrus sac, distribution of testes at the junction between proglottids, asymmetrical eggs with one pointed eggs, etc. and were identified as Spirometra erinacei(Rudolphi, 1819). The cases had the history of eating raw flesh of the snakes and these are considered the source of infection. PMID- 12891034 TI - [A case of congenital malaria] AB - A case of congenital malaria infection has been studied in a 46-day old female Korean infant. Her mother suffered from malaria infection during pregnancy in Uppervolta, Africa, and returned to Korea at the 9th month of gestation for delivery. At 39 days of age, the clinical features characterized by fever, irritability, pallor, jaundice and hepatosplenomegaly were developed. The laboratory data revealed a hemolytic anemia with thrombocytopenia, hyperbilirubinemia and increased hepatic enzyme values. A peripheral blood smear demonstrated intraerythrocytic malarial parasites snd gametocytes of Plasmodium falcifarum. She was successfully treated with quinine sulfate (25 mg/kg/day in three doses for 5 days) and trimethoprim/sulfamethoxazole (8 mg/kg/day in two doses for 5 days) orally, and repeated blood smear had been negative for malaria. This report also signifies the frst description of congenital malaria in Korea imported from Uppervolta in Africa. A brief review of related literature was made. PMID- 12891035 TI - [Infestation of larval trematodes from fresh-water fish and brackish-water fish in River Hyungsan, Kyungpook Province, Korea] AB - In order to determine the infestation patterns for metacercaria of digenetic trematodes, several kinds of fresh-water and brackish-water fish were caught in River Hyungsan, Kyungpook Province, Korea and examined from March to October in 1983. They were collected by netting, fishig with rod and line, or using the bait of crushed oil cake in transparent plastic bowls, and were dissected into the flesh, scales and fins with tail, and then examined for the presence and infestation density of larval trematodes. Of the 11 kinds of fish examined, the encysted larvae of Exorchis oviformis were found all of them, Metagonimus yokogawai from 9 kinds, Cyathocotyle orientalis, Metacercaria hasegawai and Metorchis orientalis from 6 kinds, and Clonorchis sinensis from 4 kinds in the present study. In the intensity of infestation with Clonorchis sinensis, the Korean shiner, Gnathopogon atromaculatus Nichols et Pope, was the most heavily infected species, the average number of metacercariae per gram of fish flesh being 45.5, followed by the striped shiner, Puntungia herzi Herzenstein 11.6, and Southern top-mouthed minnow, Pseudorasbora parva (Temminck et Shlegel), 9.7. The intensity of infestation with the Metagonimus larvae in 8 species of fresh-water fish was relatively low, and the average number per gram of flesh varied from 0.23 to 9.1. In comparison, the brackish-water fish were more heavily infected and the mean number of cysts per gram of flesh in the sweetfish, Plecoglossus altivelis, was 32.9. The results of this study generally indicate that Clonorchis sinensis and Metagonimus yokogawai infection may occur by consuming raw fresh water fish caught in the river Hyungsan, Kyungpook Province, Korea. PMID- 12891036 TI - [Study on the cestodes in Theragra chalcrogamma] AB - This study was performed to investigate the intestinal parasites (especially cestoda) in the guts of Theragra chalcogramma in Korea. The cestode larvae collected were morphologically observed and measured, and at the same time, microscopical study was done with the section slides of the cestode larvae. They were identified on the basis of the descriptions by Schmidt and Yamaguti. The results obtained in this study are as follows: The cestode larvae were identified as Khawia sinesis, Eulacistorhynchus chiloscyllius, Tentacularia coryphaenea, Pseudonybelinia odontacantha and Nybelinia lingualis. These are new species reported in Korea. The characteristics of Khawia sinesis are its scolex lacking loculi and broad, flat and fimbriate shape. Its neck is not separated from the body but a little constricted. Eulacistorhynchus chiloscyllius is characterized by fairly long and acraspedote scolex. The bulbs are long; retractor muscles are attached to bases of the bulbs. The two bothridia are oval or round in shape and lacking posterior notch. The tentacles are long and poeciloacanthus; double chainette present. Tentacularia coryphaenea is characterized by its long craspedote and subcylindrical scolex. The bothridia are separated, without free borders and spinous. The tentacles are short, slender, armed with solid hooks in spirals, similar except the base of tentacle. The tentacle sheaths are not twisted. The bulbs are ellipsoidal. Pseudonybelinia odontacantha is characterized by its craspedote scolex. The tentacles are inserted near anterior margin of bothridia and armed with hooks in spirals ascending left to right. The hooks are similar, throughout tentacle, with feeble curve and provided with a tooth-like protuberance on the ventral side. The bulbs are three times longer than its width. The posterior margins of bothridia have a pair of eversible ciliated pits; or fossettes. Nybelinia lingualis is characterized by its short and craspedote scolex. The bothridia are separated, with free boarders. The tentacles are cylindrical, armed with solid and similar hooks in quincunxes. PMID- 12891037 TI - A new cestode Raillietina (Skrjabinia) doggaddaensis n. sp. from Gallus gallus domesticus (L.) from India. AB - Anew poultry cestode Raillietina(Skrjabinia) doggaddaensis n.sp.has been described from the domestic fowl Gallus gallus domesticus (L.). The new species has been compared with R.(S.) dhuncheta Sharma (1943) and R.(S.) magnicoronata(Fuhrmann, 1908). PMID- 12891038 TI - On A New Reptilian Cestode Oochoristica Pauriensis N.Sp. From Hemidactylus Brooki (Gray) And Hemidactylus Flaviviridis (Ruppell) From Garhwal Hills, U.P., India. AB - Oochoristica pauriensis n.sp. has been described and figured from Hemidactylus brooki(Gray) and H. flaviviridis(Ruppell) on size of strobila, scolex, suckers, testes, ovary, eggs and oncosphere, and number of testes. The new species has been compared with close species. PMID- 12891039 TI - Collection Of Adult And Larval Mosquitoes In U.S. Army Compounds In The Republic Of Korea During 1979~1983. AB - This paper is the result of adult female mosquito and larval collection in U.S. Army Installations in Korea from 1979 to 1983. New Jersey light traps ere operated for ault collection from May to October. The primary concern of this surveillance is to determine when to recommend insecticide spraying for mosquito control in the Army areas. The 5th Preventive Medicine Unit have developed an "index" level of female mosquitoes in a light trap similar to other U.S.Army Agencies in other parts of the world. When 10 female mosquitoes are reached on two consecutive trap-night, or 5 known vector females are collected, fogging is recommended in the trap areas. Mosquito collections were conducted in 12 U.S. Army areas by operating 39 New Jersey light traps. Mosquitoes collected from the areas were identified to be 17 species comprising 3 genera. Anopheles sinensis(40%). Culex tritaeniorhyncus(31%), Aedes vexans nipponii(19%), and Culex pipiens pallens(10%) appeared to be the most common species in the area. The species, population density and monthly appearance of adult mosquitoes were found to be almost the same in the all provinces involved. And Japanese encephalitis vector mosquitoes, Culex tritaeniorhyncus, showed their seasonal fluctuation from July to September with a peak in August each year. 3. Larval habitats confirmed in the Army areas were categorized into 16 types as shown in Table The mosquito larvae collected in those habitats were identified to be 15 species representign 4 genera. Most breeding sites in the Army areas were those which are activated during the wet season. More mosquitoes were collected from the Kyungki province than from the other provinces. The reason for more collection of mosquitoes from military installations in the Kyungki provinces appears to be the geographic characteritics surrounded by rice fields, marshes and other stagnant water areas. PMID- 12891040 TI - Recent patterns of intestinal helminth infections among the residents in Taegu City, Korea. AB - In order to estimate the recent pattens of intestinal helminth infections among the residents in Taege City, Korea, a survey based on discovery of helminth eggs by formalin-ether sedimentation and modified Graham's scotch-tape anal swab techniques, were performed during the period from March to September in 1983. A total of 1,697 fecal specimens were collected from 854 male and 843 female residents from 5 villages(Dong), each of whom were selected randomly to represent 1,000 persons referred to the census in 1982. Among the specimens examined, one or more species of helminth parasites were found in 473, revealing the overall positive rate of 27.9 pre cent. Of them, Trichuris trichiura was found most frequently, in 13.2 percent, followed by Enterobius vermicularis in 7.0 per cent. Hookworm was the least prevalent. The sex-specific rate of overall helminth infections was a little higher in females than in males, while in cases of Clonorchis sinensis, Metagonimus yokogawai and Taenia species, the infection rates in males was higher than in females. T.trichiura was most prevalent in the 20~29 age group and showed about the same prevalence in males and females of all age groups. C. sinensis infection rate increased with age while those of Ascaris lumbricoides decreased. Single infections were 82.5 per cent of all positive cases. Of cource, T. trichiura revealed the highest prevalence in 31.7 per cent followed by E. vermicularis in 20.1 per cent and A. lumbricoides in 14.8 per cent. Of the double infections, the percentage of T. trichiura in combination with other intestinal helminths was higher than that of A. lumbricoides. It was concluded that although the infection rates of intestinal helminths among the residents in Taegu City, Korea is still high, eradication of these parasitic diseases seems to be possible with twice-a-year administration of specific anthelmintics, in combination with extensive public health education and improvement of the dietary life. PMID- 12891041 TI - [Epidemiological Studies On Protozoan Infection In Gyeong-Gi Do And Jeonra Bug Do] AB - The present study was designed to elucidate the recent prevalence of the intestinal protozoa and to demonstrate the relationship on prevalence of the surveyed population with their academic career and living conditions. During the period from May to July 1981, total 2,735 fecal specimens were collected from the inhabitants of 11 areas in Gyeong-gi Do (province) near by Seoul City, and 6 areas in Jeonra Bug Do, southwestern part of Korea. The samples were fixed with SAF solution and were examined by formalin-ether sedimentation technique. The results are summarized as follows: Overall positive rate of intestinal protozoa was 11.1 %: Gyeong-gi Do, 10.7 %; Jeonra Bug Do, 11.7 %. Five species of intestinal protozoa were detected: Entamoeba histolytica 0.69 %, Entamoeba coli 5.52 percent, Endolimax nana 2.71 %, Giardia lamblia 2.23 %, Iodamoeba butschlii 0.l8 %. Highest positive rates among the examined areas were recognized: Incheon City of Gyeong-gi Do, 26.7 %; Im-sil Gun(county) of Jeonra Bug Do, 19.9 %. By species, E. nana showed the highest prevalence rate (13.74 %) in Incheon City, and E. histolytica (2.37 %) and E. coli (9.95 %) in Im-sil Gun. Prevalence of E. histolytica in rural areas was higher than in urban areas. No significant difference in the prevalence rate by sex and age was recognized, although the group under 10 years and over 60 years were lower than in other age groups. The highest positive rate of E. histolytica was observed in pauper group and significant linearity of prevalence rate of E. histolytica by monthly income was observed. The inhabitants who used conventional latrine showed higher prevalence, 11.5 %, than the group of inhabitants who used modified privy. Although no significant difference of positive rate was recognized by academic career, but the lowest prevalence rate was observed in college graduates. Over half of the surveyed population, 64.8 % has no experience on anthelmintic practice. Number of anthelmintic application resulted no difference in prevalence. In conclusion, prevalences of intestinal protozoa in Gyeong-gi Do and Jeonra Bug Do decreased remarkably in comparison with those of one decade before. However, economic level, academic career and anthelmintic administration seemed no essential reasons for such declination. PMID- 12891043 TI - Seasonal Effects on Clonorchicidal Substances from Epidermal Mucus of Cyprinus carpio, Ophicephalus argus and Parasilurus asotus. PMID- 12891042 TI - [The Wormicidal Substance Of Fresh Water Fishes On Clonorchis Sinensis V. Purification And Chemical Characterization Of Clonorchicidal Substance From Epidermal Mucus Of Cyprinus Carpio] AB - As a series of studies to clarify clonorchicidal substances in body surface mucus of some fresh-water fishes, the substance in the epidermal mucus of Cyprinus carpio was isolated by silica gel column and thin layer chromatography and analyzed for its chemical nature. Wormicidal trial was done in vitro, and the results obtained are summarized as follows: The mucus was extracted by ethyl ether and separated into 4 fractions by column chromatography using benzene as solvent. The second fraction with yellowish red colour among them showed the strongest clonorchicidal effect. The yellowish red fraction obtained by column chromatography was then fractionated into 6 spots by thin layer chromatography with petrol. ether/chloroform(30/70, v/v), and the Rf. 0.714 spot among the 6 spots showed the strongest effect. The Rf. 0.714 spot was further fractionated into 6 spots by thin layer chromatography with benzene/acetone (90/10, v/v), and the Rf. 0.800 spot among the later 6 spots revealed the strongest effect. The Rf. 0.800 spot was chromatographed on column with benzene and 2 fractions were obtained. The second fraction of light brown colour represented the final purified fraction. By these purification procedures, clonorchicidal substance was purified 15-fold with 0.03 percent yield from the mucus of C. carpio, and 10mg of the final fraction killed the cercaria in 26 min, the metacercaria in 115 min, and the adult in 443 min. Infra red and nuclear magnetic resonance spectrometric analysis of the purified substance revealed that the substance belongs to an ethyl ester of unsaturated fatty acid with 2 double bonds, 15 methylene groups and l methyl group. PMID- 12891044 TI - [Status of Enterobius vermicularis Iifection in primary school children, Yongyang Kun, Kyongbuk Province] AB - An epidemiological study was undertaken to estimate the prevalence of Enterobius vermicularis infection among the primary (and infant) school children in Yongyang gun, Kyongsangbuk-do. During the period from 2 to 14 December 1983, a total of 2,227 school children in 13 schools were examined one time by means of scotch tape anal swab technique with a simple questionaire study on perianal itching, enuresis and school record. The results are as follows:The overall egg positive rate was 64.1 percent and the rate ranged from 46.6 to 86.7 percent by schools. The egg positive rates of boys and girls were 60.5 and 68.0 percent respectively and the difference was statistically significant (p<0.005). There was noted a tendency of decrease in prevalence by increase of the age of children. It was revealed that perianal itching and school record were significantly correlated with E. vermicularis infection (p<0.05) while not in case of enuresis (p>0.l). PMID- 12891045 TI - [Head Louse Infestation Among Primary School Children In Seosan-Gun, Chungnam Province] AB - The head louse infestation had been no problem since 1960s in Korea. However, the present study revealed highest infestation rate among the primary school children in Seosan-gun, Chungnam Province in July 1983. The cases of louse positive were determined by identifying the adult worms and/or their nits on scalp and hairs. The overall louse positive rate among 615 children examined was 73.5 percent. The rate was higher in girls (78.8 percent) than in boys (67.6 percent) and was highest in 3rd-4th school grades. It was observed that the more the number of family members the higher the infestation rate of children. The positive children were treated with 20 percent benzyl benzoate solution after the test for louscidal effect in petri dish. However, establishment of intensive control measure is needed to prevent further infestation. PMID- 12891046 TI - Bibliography. Current world literature. Adolescent medicine. PMID- 12891047 TI - New contraceptive methods: update 2003. AB - The decline in adolescent pregnancy rates noted in the 1990s has been attributed to more teenagers choosing abstinence, the availability of long-acting contraceptive options, and the increased use of condoms. Many adolescents remain at risk of unplanned pregnancy, however, because of method failure, inconsistent or incorrect use, or discontinuation of a method without choosing another method. Practitioners who care for adolescents and young women have access to several new methods in 2003. Recent additions include a contraceptive transdermal patch, a hormone-releasing intravaginal ring, new formulations of pills, and a new intrauterine device. Additionally, the World Health Organization has updated its guidelines to provide the practitioner with evidence-based recommendations to assist in selecting the most appropriate contraceptive method for each patient. PMID- 12891048 TI - Hypertension in adolescents: a review of diagnosis and management. AB - Long-term hypertension contributes to significant cardiovascular and renal morbidity and mortality. Although chronic hypertension is much rarer in the adolescent population than in adults, identifying the hypertensive adolescent and intervening with risk factors such as obesity that may promote hypertension is important for the clinician treating adolescents. Since both primary and secondary causes of hypertension may exist in the adolescent, a thorough and sequential clinical and diagnostic evaluation must be undertaken, including screening urinalysis, blood chemistries, and renal sonography. There are pitfalls in interpreting casual blood pressure measurements in adolescents, and the role of ambulatory blood pressure monitoring is evolving. Lifestyle modifications, including diet, exercise, and limitation of sodium intake, remain the foundation of treatment. Commonly used medications include calcium channel blockers, angiotensin receptor blockers and converting enzyme inhibitors, beta blockers, and diuretics. When considering medication in the hypertensive adolescent, potential complications of therapy must be reviewed in light of the physical and psychosocial changes ongoing in this age group. PMID- 12891049 TI - Postexposure prophylaxis: an intervention to prevent human immunodeficiency virus infection in adolescents. AB - Postexposure prophylaxis is an effective intervention to prevent transmission of the human immunodeficiency virus (HIV). Administration of antiretroviral medications within 72 hours of a percutaneous or mucous membrane exposure to HIV can prevent infection. The use of postexposure prophylaxis for occupational and perinatal HIV exposures is well established. Its efficacy for nonoccupational exposures has only recently been studied. To date in the United States, there are no national guidelines regarding postexposure prophylaxis for nonoccupational exposures. Decisions regarding its use should be made after weighing the risks of seroconversion for a given exposure, the risks of HIV in the source, and the potential toxicities of the antiretroviral medications. Several state guidelines may also serve as important resources. Clinicians should consider initiating postexposure prophylaxis in adolescents for any oral, anal, or vaginal exposure to definitely or possibly HIV-infected blood or body fluids. Adolescents treated should be followed closely for medication toxicity, acute seroconversion, and risk-reduction counseling. Further studies are needed to characterize the use and efficacy of postexposure prophylaxis in an adolescent population. PMID- 12891050 TI - Helping adolescents cope with stress during stressful times. AB - Increasing levels of stress in the daily lives of adolescents is an important health concern. Adolescents experience a variety of stressful situations and use a wide range of coping strategies to help effectively manage stress. Recent research on adolescent coping with stressful situations is reviewed. Findings support an association between coping and adolescent health problems, chronic physical illness, and mental health. More research is needed to investigate the long-term benefits of coping with stress on adolescent health and well-being. Specific recommendations for helping adolescents cope with stressful situations are discussed. Health care providers are encouraged to assess how adolescents cope with acute and chronic stressors and provide adolescents with information about coping with stressful situations. PMID- 12891051 TI - Sexually transmitted diseases treatment guidelines. AB - Sexually transmitted diseases (STDs) are a major health problem for adolescents. Health care providers for adolescents play a critical role in preventing and treating STDs. In May 2002, the Centers for Disease Control and Prevention published the Sexually Transmitted Diseases Treatment Guidelines 2002. These evidence-based guidelines are based on a systematic literature review focusing on information that had become available since the 1998 Guidelines for Treatment of STDs. This article reviews the new STD treatment guidelines for gonorrhea, chlamydia, bacterial vaginosis, trichomonas, vulvovaginal candidiasis, pelvic inflammatory disease, genital warts, herpes simplex virus infection, syphilis, and scabies. Although these guidelines emphasize treatment, prevention strategies and diagnostic recommendations also are discussed. PMID- 12891052 TI - Bibliography. Current world literature. Dermatology. PMID- 12891053 TI - Infection in atopic dermatitis. AB - PURPOSE OF REVIEW: Atopic dermatitis is a chronic inflammatory skin disease commonly seen in children, but it also occurs in adults. This skin disease is often triggered by bacterial, fungal, or viral skin infections. The mechanisms resulting in this increased propensity for skin infections have been an area of active investigation. RECENT FINDINGS: Recent studies suggest that the skin of patients with atopic dermatitis has increased avidity for binding to Staphylococcus aureus and is deficient in its ability to generate antimicrobial peptides needed to eradicate infectious agents. SUMMARY: The current review will examine recent literature on the etiology and pathogenesis of infection in atopic dermatitis. These observations are likely to have importance implications for management of atopic dermatitis. PMID- 12891054 TI - Update on pathogenesis and treatment of acne. AB - Acne vulgaris is a common cutaneous disorder of the pilosebaceous follicle. More than 45 million people are affected with acne vulgaris. In addition, 80% of people in the United States report that they suffered from acne at one point in life. It is estimated that US consumers spend $1.2 billion each year for the treatment of acne. Typically, lesions are pleomorphic and range from open and closed comedones to inflammatory papules, pustules, cysts, and nodules, and scarring may result. The disease is important, with a significant effect on patients' self-esteem. The pathogenesis is only partially understood and is multifactorial. Successful management of acne requires an understanding of the pathophysiology of disease. The first half of this review discusses new discoveries in the pathogenesis of acne, and the second half introduces new and time-honored therapies. PMID- 12891055 TI - Type 2 diabetes: an epidemic disease in childhood. AB - Type 2 diabetes mellitus in the pediatric population is now a public health problem. It represents 8 to 45% of all diabetes reported among children and adolescents. Concurrently, childhood obesity has become an epidemic in the United States. Epidemiologic risk factors and diagnostic criteria have largely been established: The incidence of type 2 diabetes increases with age and obesity. Children and adolescents diagnosed with type 2 diabetes usually have a first- or second-degree relative with the disorder. Familial clustering and twin studies have suggested a genetic component for type 2 diabetes. However, the molecular mechanisms that promote diabetes in susceptible individuals, the best treatment plans, and methods of prevention of this disorder are not yet established. PMID- 12891056 TI - Thyroid hormone deiodinases: physiology and clinical disorders. AB - Thyroid hormone action is achieved through the binding of 3,5,3'-triiodothyronine to its nuclear receptor, which results in alterations in gene expression. An impairment in thyroid hormone action during vertebrate development results in severe, irreversible abnormalities in tissue growth, maturation, and function. The deiodinases are a family of selenoproteins expressed in a number of fetal and adult tissues that catalyze the activation and inactivation of thyroid hormones. Their unique biochemical characteristics and tissue and developmental expression patterns suggest that deiodinases may control the concentration of active thyroid hormone available to specific tissues or cell types at certain stages of development. The deiodinases thus appear to play an important role in regulating thyroid hormone action at a prereceptor level. Current research focusing on a better understanding of the biochemistry, regulation, and physiologic role of these enzymes is the focus of this review. PMID- 12891057 TI - Immunizations, neonatal jaundice, and animal-induced injuries. AB - This article reviews recent advances and recommendation changes in the areas of immunization, neonatal hyperbilirubinemia, and animal-induced injury. Physician education of families and care of patients in these three topic areas are important public health measures. First, pediatricians are always actively promoting public health through immunization, so the 2003 vaccination recommendations are critiqued along with perceived barriers to proper immunization. New information about smallpox vaccine is included in this year's review because of increased concerns about the use of smallpox as a weapon of bioterrorism since the world events in recent years. Next, the continued study of the management of hyperbilirubinemia in preventing kernicterus is examined. Finally, the management and prevention of animal-induced injuries is reviewed. In each of these areas, the pediatrician plays a prominent role in promoting the health and well being of children. PMID- 12891058 TI - Human herpesvirus 8 epidemiology: what we do and do not know. PMID- 12891059 TI - Dendritic cells generated in the presence of granulocyte-macrophage colony stimulating factor and IFN-alpha are potent inducers of HIV-specific CD8 T cells. AB - OBJECTIVE: To investigate the ability of granulocyte-macrophage colony stimulating factor (GM-CSF) and IFN-alpha to induce the differentiation of peripheral monocytes into dendritic cells (DC) and their ability to trigger an HIV-specific CD8 T-cell response. METHODS: Monocytes isolated from both seronegative controls and HIV-infected individuals were differentiated into DC using GM-CSF with either IL-4 or IFN-alpha for 7 days. We assessed the phenotypic characteristics and IL-12 production by flow cytometry. The ability of DC to trigger CD8 T-cell responses was assessed by means of ELISpot and cytotoxicity assays. In addition, HIV-1-RNA levels were measured in culture supernatants. RESULTS: Compared with control DC generated in the presence of GM-CSF and IL-4, DC generated in the presence of GM-CSF and IFN-alpha expressed higher levels of MHC class I molecules and produced similar or higher levels of IL-12 after CD40 ligation or Staphyloccus aureus Cowan stimulation. GM-CSF/IFN-alpha DC expressed low levels of CD4, CXCR4 and DC-SIGN and did not produce detectable virus during the differentiation period. Pulsed GM-CSF/IFN-alpha DC were found to prime CD8 T cells from HIV-negative controls to exert cytotoxic activity against target cells expressing HIV antigens. HIV peptide-pulsed GM-CSF/IFN-alpha DC promote specific IFN-gamma production by autologous CD8 T cells from HIV-seronegative donors. Furthermore, GM-CSF/IFN-alpha DC from HIV-seropositive patients efficiently present HIV peptides to autologous CD8 T lymphocytes. CONCLUSION: GM-CSF and IFN alpha allow the generation of DC with high CD8 T-cell stimulating abilities. Therefore, this strategy may represent a novel approach to therapeutic vaccination in HIV disease. PMID- 12891060 TI - Virological rebound after suppression on highly active antiretroviral therapy. AB - OBJECTIVE To determine the rate of virological rebound and factors associated with rebound among patients on highly active antiretroviral therapy (HAART) with previously undetectable levels of viraemia. DESIGN An observational cohort study of 2444 patients from the EuroSIDA study. METHODS Patients were followed from their first viral load under 400 copies/ml to the first of two consecutive viral loads above 400 copies/ml. Incidence rates were calculated using person-years of follow-up (PYFU), Cox proportional hazards models were used to determine factors related to rebound. RESULTS Of 2444 patients, 1031 experienced virological rebound (42.2%). The incidence of rebound decreased over time; from 33.5 in the first 6 months after initial suppression to 8.6 per 100 PYFU at 2 years after initial suppression (P < 0.0001). The rate of rebound was lower for treatment naive compared with treatment-experienced patients. In multivariate models, patients who changed treatment were more likely to rebound, as were patients with higher viral loads on starting HAART. Treatment-naive patients were less likely to rebound. Among pretreated patients, those who were started on new nucleosides were less likely to rebound. CONCLUSION The rate of virological rebound decreased over time, suggesting that the greatest risk of treatment failure is in the months after initial suppression. Treatment-naive patients were at a lower risk of rebound, but among drug-experienced patients, those who added new nucleosides had a lower risk of rebound, as were patients with a good immunological response. PMID- 12891062 TI - Patterns of adherence to antiretroviral medications: the value of electronic monitoring. AB - OBJECTIVE: To investigate the patterns of intra-subject (between medication) adherence to antiretroviral therapy. DESIGN: A prospective, observational, 3 month study of adherence to antiretroviral therapy at an inner-city clinic in 40 HIV-infected subjects. METHODS: Adherence was monitored monthly by the use of medication event monitoring system (Aprex) caps placed on each antiretroviral drug in a subject's regimen. Agreement between different drug classes and dosing schedules, for each subject, was quantified by estimating the mean difference in adherence, with 95% limits of agreement. An analysis of variance model was used to estimate the variance of the differences. Individual dosing calendars were examined for each subject. RESULTS: The dosing schedule was a strong predictor of intra-subject adherence. Regardless of the subject's overall adherence rate, high or low, when subjects missed a dose of one medication, they missed a dose of both medications taken at that dosing time. Conversely, when medications were scheduled to be taken together, regardless of the drug class, the medications were taken at the same times. The majority of the subjects took medications at obviously incorrect times. Problematical adherence was related to thrice-daily dosing and food restrictions. CONCLUSION: This is the first report objectively to quantify intra-subject adherence to antiretroviral therapy and report the findings in detail. We observed clear patterns of drug-taking behavior among the subjects in our study. To the extent that medication scheduling is a controllable factor, our report provides an insight into specific patterns of behavior that may be targets for adherence counseling. PMID- 12891061 TI - Expression of adipogenic transcription factors, peroxisome proliferator-activated receptor gamma co-activator 1, IL-6 and CD45 in subcutaneous adipose tissue in lipodystrophy associated with highly active antiretroviral therapy. AB - OBJECTIVE: To determine the expressions of multiple genes in the subcutaneous adipose tissue of HIV-positive, highly active antiretroviral therapy (HAART) treated patients with and without lipodystrophy. DESIGN AND METHODS: Real-time polymerase chain reaction was used to measure gene expressions in this cross sectional study. RESULTS: The messenger RNA concentrations of adipose transcription factors (peroxisome proliferator-activated receptor (PPAR) gamma and delta and sterol regulatory element binding protein 1c) were all significantly lower in the lipodystrophic than the non-lipodystrophic group. The mRNA concentration of PPAR-gamma co-activator 1 (PGC-1), which regulates mitochondrial biogenesis, was lower in the lipodystrophic than the non lipodystrophic group. The mRNA expression of lipoprotein lipase, acyl coenzyme A synthase and glucose transport protein 4 were significantly lower in the lipodystrophic than the non-lipodystrophic group, but the mRNA concentrations of fatty acid transport and binding proteins were similar in both groups. The mRNA concentrations of IL-6 and CD45 (a common leukocyte marker) were significantly higher in the lipodystrophic than the non-lipodystrophic group. CONCLUSION: Multiple alterations characterize gene expression in the subcutaneous adipose tissue of patients with HAART-associated lipodystrophy compared with HIV positive, HAART-treated patients without lipodystrophy. The low expression of transcription factors inhibits adipocyte differentiation. The low expression of PGC-1 may contribute to mitochondrial defects. In addition, IL-6 and CD45 expressions are increased, the latter implying an excessive number of cells of leukocyte origin in lipodystrophic adipose tissue. Mitochondrial injury and an excess of proinflammatory cytokines may lead to increased apoptosis. All these changes may contribute to the loss of subcutaneous fat in HAART-associated lipodystrophy. PMID- 12891063 TI - Persistent mitochondrial dysfunction in HIV-1-exposed but uninfected infants: clinical screening in a large prospective cohort. AB - BACKGROUND: Antiretroviral prevention of mother to child HIV-1 is established but tolerance remains to be assessed. AIM To determine the risk for persistent mitochondrial dysfunction in HIV-uninfected children born to seropositive mothers. METHOD: An exhaustive study in a large prospective cohort with predetermined algorithm of the unexplained symptoms compatible with mitochondrial dysfunction. A total of 2644 of 4392 children were exposed to antiretrovirals. Complementary investigations were carried out on a case-by-case basis using classification with a diagnostic probability scale, based on experience with constitutional diseases. A spontaneous notification register for children not included in the cohort was created. RESULTS: Good circumstantial evidence of mitochondrial dysfunction was found for twelve children. Seven were from the cohort. All presented neurological symptoms, often associated with abnormal magnetic resonance image (10 of 12) and/or a significant episode of hyperlactatemia (seven of 12). All had either a profound deficit in one of the respiratory chain complexes (11 of 12) and/or a typical histological pattern (two of 12). All were perinatally exposed to antiretrovirals. None of them had perinatal morbidity that could explain this symptomatology. The 18-month incidence was 0.26% (95% confidence interval, 0.10-0.54) in exposed children, in comparison with the general figure of 0.01% for paediatric neuro-mitochondrial diseases in the general population. Fourteen other children in the cohort, all exposed to antiretrovirals, had unexplained symptoms, mostly neurological, for which one of the possible differential diagnoses was mitochondrial dysfunction. Close similarities in clinical, neuroradiological and histological findings strongly suggest a common pathological process in all these 26 children. CONCLUSION: Children exposed to nucleoside analogues during the perinatal period are at risk of a neurological syndrome associated with persistent mitochondrial dysfunction. PMID- 12891064 TI - Survival is prolonged by highly active antiretroviral therapy in AIDS patients with primary central nervous system lymphoma. AB - OBJECTIVE: To determine the effects of highly active antiretroviral therapy (HAART) on survival in AIDS-related primary central nervous system lymphoma (PCNSL). METHODS: Survival in consecutive patients with PCNSL at a large county teaching hospital from 1995 to 2001 were analyzed by the log rank test and Cox proportional hazards ratios (HR) were calculated for factors potentially affecting survival. RESULTS: During the study period, 25 patients were diagnosed with PCNSL: 19 definite and 6 probable. At diagnosis, median CD4 cell count was 12 x 10(6) cells/l (range 1-151) and median HIV viral load was 5.3 log(10) copies/ml (range 3.9-5.9). Sixteen patients died (median survival 87 days; range, 0 to > 2112). Longer survival was noted for patients who received HAART after diagnosis [HR for death, 0.06; 95% confidence interval (CI), 0.01-0.48]. Six of seven HAART-treated patients were alive versus 0/18 untreated patients at a median follow-up time of 667 days (P = 0.0007 by log rank test). A survival benefit was seen for patients who had >/= 0.5 log(10) copies/ml decrease in HIV viral load after diagnosis (n = 6; HR, 0.07; 95% CI, 0.01-0.55) and for patients with a significant CD4 cell rebound (increase >/= 50 x 106 cells/l) in response to HAART (n = 6): all survived versus 0/19 survived (P = 0.0003). Cranial radiation therapy (n = 13) prolonged survival (HR, 0.20; 95% CI, 0.07-0.58). Median survival was only 29 days for 11 patients who received neither radiation nor HAART. CONCLUSIONS: Receipt of HAART after diagnosis is associated with a significantly longer survival in patients with AIDS-related CNS lymphoma. PMID- 12891065 TI - Clinically relevant interpretation of genotype for resistance to abacavir. AB - OBJECTIVE: To develop a stepwise methodology for the development and validation of clinically relevant genotypic score for resistance to antiretroviral drugs and to apply this approach to the genotypic resistance to abacavir. METHODS: All patients having received abacavir during the Narval trial were included in this study. The impact of each nucleoside analogue resistance mutation on the virologic response to abacavir was studied in a univariate analysis. Mutations with a P value < 0.20 and those selected by abacavir were retained. According to the number of mutations three levels of resistance were defined. A multivariate analysis accounting for confonding variables assessed whether the genotypic score was an independent predictor of the response. The robustness of the score was analysed using the bootstrap resampling method. RESULTS: In the 175 patients exposed to abacavir, the strongest association between the decrease in viral load and the number of mutations was observed with a set of six mutations at codons 41, 67, 210, 215, 74 and 184 of the reverse transcriptase gene. In patients with fewer than four mutations (no evidence of resistance) the median decrease in viral load was -1.64 log(10) copies/ml while it was -0.69 log(10) and -0.19 log(10) in those with four (possible resistance) and five or six (resistance) mutations respectively. In the multivariate analysis this score was an independent predictor of the response. The bootstrap analysis showed the robustness of the score. CONCLUSIONS: We developed a new strategy for the analysis of correlation between genotype profile at baseline and virologic response. PMID- 12891066 TI - Mortality due to hepatitis C-related liver disease in HIV-infected patients in France (Mortavic 2001 study). AB - OBJECTIVE: To determine mortality due to end-stage liver disease (ESLD) in a nationwide cohort of HIV-infected patients 5 years after the introduction of highly active antretroviral therapy (HAART) and to compare this with that observed before and during the early years of HAART. DESIGN: and methods: All departments of internal medicine and infectious diseases from the GERMIVIC Study Group prospectively recorded all deaths in HIV-infected patients during 2001. Sixty-five departments, following a total of 25 178 HIV-infected patients, participated in the study. Results were compared with those of previous surveys conducted using similar methodology in 1995 and 1997. RESULTS: Among 265 deaths observed during 2001, 129 (48.7%) were related to AIDS, 38 (14.3%) to ESLD, and 98 (36.7%) to other causes. Mortality due to ESLD represented 28% of non AIDS related deaths; 36 of the 38 patients (95%) dying from ESLD had chronic hepatitis C virus (HCV) infection. In 2001, deaths due to ESLD (14.3%) were significantly more frequent than in 1995 (1.5%; P < 0.01) and 1997 (6.6%; P < 0.01). During this interval, the prevalence of hepatocellular carcinoma as a cause of death increased (1995, 4.7%; 1997, 11%; 2001, 25%; P < 0.05), as did alcohol consumption (P < 0.01). CONCLUSIONS: In the post-HAART era, ESLD due to HCV is a growing cause of mortality in HIV-infected patients. Increased longevity attributable to HAART, and a higher prevalence of alcohol consumption, are probably involved in this trend. PMID- 12891067 TI - Prevalence of HIV and other sexually transmitted infections, and risk behaviours in unregistered sex workers in Dakar, Senegal. AB - OBJECTIVES: To estimate the prevalence rates of HIV and other sexually transmitted infections (STI) among unregistered sex workers, and to describe their sociodemographic characteristics and sexual behaviours, and the reasons why they were not officially registered as sex workers, in order to design specific public health interventions. METHODS: A one-stage cluster-sample survey was conducted in Dakar in 2000. Unregistered sex workers were interviewed in randomly selected establishments (official and clandestine bars, brothels and nightclubs), and blood, endocervical and vaginal samples were collected for laboratory diagnosis. RESULTS: A total of 390 women with a median age of 29 years were recruited. One-seventh of them were under the legal age for prostitution in Senegal (21 years). The median length of prostitution was 24 months and 73.5% of the women stated regular prostitution. Three-quarters of the women were found to have markers for at least one infection. The prevalence rates were as follows: HIV-1, 6.0%; HIV-2, 3.6%; HIV-1+2, 0.4%; syphilis, 23.8%; gonorrhea, 22.0%; chlamydial infection, 20.0%; trichomoniasis, 22.4%; candidiasis, 19.0%; and bacterial vaginosis, 28.8%. The main reported reason for non-registration was ignorance of the legal system and its procedures (19.4%); 18.9% of the women refused to register. One-third of the women reported that their clients used condoms inconsistently or never. CONCLUSION: This survey suggests that a multidimensional public health response is needed in Senegal, comprising legal information, downwards revision of the legal age for prostitution, and specific medical follow-up based on education, condom promotion and management of STI for non-registered sex workers. PMID- 12891068 TI - Trends and measurement of HIV prevalence in northern Malawi. AB - BACKGROUND: Most data on HIV prevalence in Malawi come from antenatal clinic (ANC) surveillance and are, therefore, subject to bias. OBJECTIVES: HIV prevalence and risk factors were measured using population-based data to assess the accuracy of ANC surveillance and changes in prevalence and risk factors for HIV over time. METHODS: HIV prevalence was measured in 1988-1993 and 1998-2001 in community controls from case-control studies of mycobacterial disease in Karonga District, Malawi. ANC surveillance studies in the district began in 1999. RESULTS: Age and area-standardized HIV prevalence in women aged 15-49 years in the community was 3.9% in 1988-1990, 12.5% in 1991-1993 and 13.9% in 1998-2001. For men, HIV prevalence was 3.7%, 9.2% and 11.4% in the same periods. In 1988 1993, HIV positivity was associated with occupations other than farming, with increased schooling and being born outside Karonga District. In 1998-2001, non farmers were still at higher risk but the other associations were not seen. The age- and area-adjusted HIV prevalence in the ANC in 1999-2001 was 9.2%. The underestimate can be explained largely by marriage and mobility. Reduced fertility in HIV-positive individuals was demonstrated in both ANC and community populations. A previously recommended parity-based adjustment gave an estimated female HIV prevalence of 15.0%. CONCLUSIONS: HIV prevalence has increased and continues to be higher in non-farmers. The increase is particularly marked in those with no education. ANC surveillance underestimated HIV prevalence in the female population in all but the youngest age group. Although there were differences in sociodemographic factors, a parity-based adjustment gave a reasonable estimate of female HIV prevalence. PMID- 12891069 TI - Child survival in relation to mother's HIV infection and survival: evidence from a Ugandan cohort study. AB - OBJECTIVE: To analyse the contribution of maternal survival and HIV status to child (under-5 years) mortality in a rural population cohort in South-west Uganda. METHODS: Approximately 10 000 people residing in 15 neighbouring villages were followed between 1989 and 2000 using annual censuses and serological surveys to collect data on births, deaths, and adult HIV serostatus. Mother-child records were linked, child mortality risks (per 1000 births) and hazard ratios (HRs) for child mortality according to maternal HIV serostatus were computed, allowing for time-varying covariates. RESULTS: A total of 3727 children were born, of whom 415 died during 14 110 child years of follow-up. Mother's HIV status at birth was ascertained unambiguously for 3004 children, of whom 218 were born to HIV positive mothers. Infant mortality risk was higher for HIV seropositive than seronegative mothers (225 versus 53) as was child mortality risk (313 versus 114). Child mortality risk was also higher for mothers who died (571) than for surviving mothers (128). After controlling for child's age and sex, independent predictors of mortality in children were: mother's terminal illness or death (HR = 3.8); mother being HIV positive (HR = 3.2); child being a twin (HR = 2.0); teenage motherhood (HR = 1.7) and maternal absence (HR = 1.7). CONCLUSION: Maternal survival and HIV status are strong predictors of child survival. The higher mortality in HIV-infected women compounds mortality risks for their children, regardless of children's HIV status. Programmes aimed at the welfare of children should take into account the independent effect of mothers' HIV and vital status. PMID- 12891070 TI - HIV prevalence in 72 000 urban and rural male army recruits, Ethiopia. AB - BACKGROUND: The recent estimate of around 2 million HIV-infected people in Ethiopia derives from data that are sparse, especially in the rural areas where the majority (> 85%) of the population lives. We assessed HIV prevalence in almost 72 000 army recruits who resided in urban and rural areas prior to recruitment. METHODS: Rapid HIV tests, HIVSPOT and Determine, were conducted on blood samples drawn at enrolment from almost 10 000 urban recruits, in 1999, and 62 000 rural recruits, in 2000. Socio-demographic data from recruits were available. RESULTS: In urban recruits, overall HIV prevalence was 7.2%, ranging from 4.3 to 10.5% depending on region. In rural recruits, overall HIV prevalence was 3.8%, but the majority were farmers (57%) and students (18%) with an HIV prevalence of 2.7% and 2.6%, respectively. (Higher) level of education in rural recruits was associated with HIV infection. Rural recruits of the Muslim religion were less likely to be HIV infected than recruits of the Orthodox Christian religion (odds ratio: 0.7; 95% confidence interval, 0.65-0.84). Urban and rural residents of Amhara region were at higher risk of HIV infection. CONCLUSION: Prevalence in both rural and urban army recruits is below previous estimates. Geographic distribution of HIV is uneven. The impact of religion, education, and region on HIV prevalence suggests avenues for targeting HIV prevention efforts in Ethiopia. PMID- 12891071 TI - Regional differences in HIV trends in The Gambia: results from sentinel surveillance among pregnant women. AB - OBJECTIVE: To monitor HIV-1 and HIV-2 trends in The Gambia, West Africa. METHODS: In 1993-1995 a nationwide survey among 29 670 pregnant women attending eight antenatal clinics estimated the seroprevalence of HIV-1 at 0.6%, and of HIV-2 at 1.1%. Five years later, sentinel surveillance in pregnant women was established, using unlinked anonymous testing in four clinics. A dried blood spot on filter paper was obtained and tested for HIV antibodies. RESULTS: Between May 2000 and August 2001, 8054 analysable samples were collected at four sites. The prevalence of HIV-1 rose sharply in one rural area from 0.6 to 3.0% (P < 0.0001), but the increase was small and non-significant in two other rural sites and in the urban site. The prevalence of HIV-2 did not change significantly at any of the sites. The overall prevalence of HIV-1 was 1.0% [95% confidence interval (CI) 0.8-1.3%], and of HIV-2 0.8% (CI 0.6-1.0%). Site, nationality and higher age were significantly associated with HIV-1 infection, and higher parity and site were significantly associated with HIV-2 infection. CONCLUSION: Fifteen years after the first case of HIV-1 was described in The Gambia, the epidemic is still at a low level. There is heterogeneity within the country, with one rural area experiencing a fivefold increase in 6 years. The prevalence of HIV-2 in The Gambia is stable. PMID- 12891072 TI - Detection and quantification of Kaposi's sarcoma-associated herpesvirus to predict AIDS-associated Kaposi's sarcoma. AB - OBJECTIVE: To identify immunologic and virologic predictors of AIDS-associated Kaposi's sarcoma (KS). DESIGN: Nested case-control analysis of KS risk in a cohort of 132 HIV-infected homosexual men in New York and Washington, DC, USA. METHODS: For each KS case, we selected two HIV-infected controls, matched for CD4 cell count and Kaposi's sarcoma-associated herpesvirus (human herpesvirus 8; KSHV) serostatus (enzyme immunoassay for antibody to KSHV protein K8.1). Cell associated KSHV and Epstein-Barr virus (EBV) viral loads were measured with quantitative real-time PCR assays on samples collected 1 year (median) before KS diagnosis. RESULTS: Thirty-one men developed AIDS-associated KS (incidence 3.1 per 100 person years). Among HIV-infected men, KS incidence was higher among those with K8.1 seropositivity (5.0 versus 1.4 per 100 person years; P = 0.004), low CD4 cell count [hazard ratio (HR), 1.49; 95% confidence interval (CI), 1.24 1.79 per 100 x 10(6) cells/l decline), or high HIV RNA level (HR, 3.96; 95% CI, 2.19-7.16 per log(10)). In the case-control analysis, nine of 70 evaluated subjects had KSHV viremia, generally low level (median viral load 180 copies per 1 x 10(6) cells). KSHV viremia was associated with increased KS risk (unadjusted odds ratio, 9.1; 95% CI, 1.7-48; odds ratio, 11.7; 95% CI, 1.8-76 after adjustment for K8.1 serostatus, CD4 cell count, and HIV RNA). Among K8.1 seropositive subjects, KS incidence was tenfold higher in those with KSHV viremia (30.3 per 100 person years versus 3.4 per 100 person years in those without viremia). Also, EBV viral loads were higher in cases than in controls (P = 0.07). CONCLUSIONS: Among individuals with HIV-KSHV coinfection, KSHV viremia identifies a subgroup with extremely high risk for developing KS. PMID- 12891073 TI - Vertical transmission of HIV in Rio de Janeiro, Brazil. AB - A cohort of 297 HIV-infected pregnant women was followed from January 1996 to December 2001. The overall transmission rate was 3.57% and remained constant over time. Low birth-weight was independently associated with a higher risk of vertical transmission (P=0.0072), whereas a longer duration of antiretroviral drugs during pregnancy was independently associated with a lower risk of transmission (P=0.0084). Further decreases in transmission should be obtained by initiating prophylaxis earlier in pregnancy. PMID- 12891074 TI - Comparison of metabolic abnormalities 48 weeks after switching from highly active antiretroviral therapy containing a non-nucleoside reverse transcriptase inhibitor to Trizivir versus continued highly active antiretroviral therapy. AB - Forty patients participating in the TRIZAL study were treated with a non nucleoside reverse transcriptase inhibitor (NNRTI)-containing regimen before being randomly assigned either to continue their baseline therapy or to switch to a triple nucleoside regimen. No difference was observed in treatment efficacy between the two groups, and total cholesterol was observed to improve significantly in the switch group. Switch maintenance may be an appropriate strategy in patients treated with an NNRTI. PMID- 12891075 TI - Anterior neck fat deposition in lipodystrophy syndrome; a new variant on a theme? PMID- 12891077 TI - M36I, protease gene, HIV-1: resistant mutation or genetic polymorphism? PMID- 12891078 TI - A starting point to a better detection of active and latent tuberculosis infection in HIV-positive individuals. PMID- 12891079 TI - Technology can't replace compassion in health care. PMID- 12891080 TI - Atrial fibrillation: beyond "irregularly irregular". PMID- 12891081 TI - The latest in cardiac nursing. PMID- 12891082 TI - The latest in trauma nursing. PMID- 12891083 TI - The latest in respiratory nursing. PMID- 12891084 TI - ACLS pointers: acute pulmonary edema, hypotension, and shock algorithm. PMID- 12891085 TI - What can you give back to nursing? PMID- 12891086 TI - How animal-assisted therapy affects discharge teaching: a pilot study. PMID- 12891087 TI - Different ways of being aware of a psychiatric disability: a multifunctional narrative approach to insight into mental disorder. AB - Many people with a psychiatric disability appear to exhibit varying degrees of lack of awareness of the mental disorder that is implied by their diagnosis. Furthermore, there is considerable controversy regarding the significance of being aware of and acknowledging a psychiatric disorder. Different responses to the issues underlying this controversy can be found in the professional and research literature. This article describes and compares some of these responses. On the basis of this description and comparison, a multifunctional narrative approach to insight into mental disorder is developed, and implications of this approach for psychiatric rehabilitation research and for psychiatric rehabilitation intervention are explored. PMID- 12891088 TI - Paranoia in a nonclinical population of college students. AB - The present study examined the incidence of paranoid ideation in a nonclinical population. A sample of 324 college students completed a questionnaire assessing their personal experiences of paranoia, with an emphasis on the cognitive, behavioral, and affective components of their experience. They also completed a general measure of paranoia in nonclinical samples, the Fenigstein and Vanable Paranoia Scale, and the Rosenberg Self-Esteem Scale. A total of 153 participants reported an experience of paranoia, which included a clear statement of planned intention to harm. This group scored significantly higher on the Paranoia Scale than those who reported no experience of paranoia. Furthermore, greater levels of paranoid ideation were associated with lower self-esteem. The present findings suggest that paranoia is a common human experience, and are consistent with the idea of continuity between normal and abnormal experience. PMID- 12891089 TI - Belief in exposure to terrorist agents: reported exposure to nerve or mustard gas by Gulf War veterans. AB - September 11 brought increased awareness that even the threat of chemical and biological terrorism can overwhelm this country's health care system. Belief in exposure to toxic agents, even when none is documented, is not uncommon in crisis and merits vigilant health care evaluation and services. This study examined risk factors (demographics, physical symptoms, clinical diagnosis, exposures, and health status) for belief in exposure to potential terrorist agents (nerve or mustard gas) using a large sample of Gulf War veterans who reported belief in exposure to nerve or mustard gas. We found that females, nonwhites, and those who were older (age 32 to 61 years) were more likely to report exposure. When adjusting for demographics and military service, these veterans reported more exposures (nonnerve or mustard gas) to potentially toxic agents and traumatic events (odds ratio [OR], 6.80; p<.001), reported more physical symptoms during the Gulf War (OR, 2.38; p<.001), were more likely to be diagnosed with a mental disorder (OR, 1.72; p<.001), and reported poorer current health status (OR, 3.47 to 1.22; p<.001). Not unlike previously reported studies of disasters, traumatic exposures, or risk exposures, belief in exposure to toxic agents suggests that certain people are at a greater health care risk. This knowledge will aid in better responding to rapid demands that may be placed on our health care delivery systems in times of potential terrorist activity. PMID- 12891090 TI - Eating disorders and suicide risk factors in adolescents: an Italian community based study. AB - In a mixed male-female sample of 1000 adolescents age 15 to 19 years in a northeastern area of Italy, we investigated the links between eating disorders and suicidal tendencies by means of self-compiled measures, including the Eating Attitudes Test (EAT), the Bulimic Investigatory Test of Edinburgh (BITE), the Body Attitudes Test (BAT), and the SCL-90-R. More females than males reported abnormal eating patterns suggesting eating disorders: we found that 100 females (15.8%) and 8 males (2.8%) achieved scores above the suggested cutoff on EAT (cutoff = 30), 26 females (4.1%) and 1 male (.3%) achieved scores above the suggested cutoff on BITE (cutoff = 20), and 287 females (45.5%) and 24 males (8.6%) achieved scores above the suggested cutoff on BAT (cutoff = 36). More females than males also reported symptoms of hopelessness (44.3% vs. 30.5%) and suicidal ideation (30.8% vs. 25.3%). Both males and females reporting suicidal ideation achieved significantly higher scores on the eating disorders inventories, with no independent contribution by age, socioeconomic status, or body mass index. Although caution is required when drawing conclusions from self reported measures, studies on mood disorders and suicidality in youth clearly need to include measures of eating disorders. PMID- 12891091 TI - Hopelessness and suicidal ideation in outpatients with treatment-resistant depression: prevalence and impact on treatment outcome. AB - Depression and hopelessness are risk factors for suicide. The purpose of this study was to examine the extent of suicidal ideation and hopelessness in outpatients with treatment-resistant depression (TRD) and to study the impact of suicidal ideation and hopelessness on treatment with nortriptyline (NT). The degree of suicidal ideation and hopelessness was assessed during the screen visit with the use of items #3 and #30 of the Hamilton Depression Rating Scale (HAM-D) in 89 patients with TRD who entered a 6-week open trial of NT. Forty of these patients also completed the Beck Hopelessness Index (BHI) during the screen visit. In separate logistic regressions, the scores from the BHI and the two HAM D items were then tested as predictors of clinical response to the 6-week trial with NT, controlling for the severity of depression. More than half of patients reported thoughts or wishes of death to self and significant hopelessness. A greater degree of hopelessness before treatment in completers, reflected by the score on the HAM-D item #30, predicted response to NT. More than half of patients with prominent hopelessness who completed the trial responded. Patients with TRD are more likely than not to report prominent suicidal ideation and hopelessness. Furthermore, a full 6-week trial of NT, a relatively noradrenergic tricyclic antidepressant, may be particularly useful in patients who have failed to respond to several antidepressants and also report significant hopelessness. PMID- 12891093 TI - Clinical symptomatology of posttraumatic stress disorder-diagnosed Australian and United States Vietnam combat veterans: an MMPI-2 comparison. AB - The authors compared MMPI-2 scores of 95 Australian and 96 US Vietnam combat veterans diagnosed with posttraumatic stress disorder (PTSD) from structured PTSD clinical interviews. Groups were strikingly similar on the MMPI-2 clinical and validity scales but were different on two content scales, with higher scores on FRS (fears) and BIZ (bizarre mentation) for the US sample. Employment status was included as a factor, because it too discriminated groups, but it did not interact with the veteran group variable to produce scale differences. The roles of employment status and disability payments are considered in accounting for differences in the psychiatric presentations of the groups. Results suggest that American and Australian Vietnam combat PTSD samples are very similar to each other, with implications for the treatment outcome literature. PMID- 12891092 TI - Studying Chinese suicide with proxy-based data: reliability and validity of the methodology and instruments in China. AB - This study examined the reliability and validity of the instruments as used in the psychological autopsy method in China. With data from 130 informants on 66 completed suicides and 130 informants on 66 normal community controls and 66 controls themselves, the validity was examined by comparing the responses of informants and the responses (gold standards) of the target participants in the control group. All the tested instruments were shown to be reliable, and proxy respondents were generally good judges of targets' suicidal intention, social support, depression, life events, personality traits, and mental disorders. Additionally, interrater reliabilities of the five interviewers were very good on selected scales. This study has laid a partial foundation for future psychological autopsy projects to be held in Chinese culture. PMID- 12891094 TI - High psychiatric comorbidity in spasmodic torticollis: a controlled study. AB - Disturbed body image and negative self-referent cognitions caused by the postural disfigurement of the head are regarded as the main reason for elevated depression scores in spasmodic torticollis (ST), but this factor was never controlled for. We therefore compared 48 patients with ST and 48 patients with alopecia areata (AA) who were matched for age, sex, and body image dissatisfaction. Psychiatric diagnoses were based on a structured psychiatric interview (SCID-I). Results of patients with ST and AA were compared with a matched sample of the representative German population. Odds ratios to develop psychiatric comorbidity for patients with ST compared with patients with AA were significantly increased throughout nearly all assessed DSM-IV categories. Logistic regression analysis showed that (1) depressive coping and (2) belonging to the group of patients with ST correlated with a significantly higher rate of current psychiatric diagnosis. We conclude that high psychiatric comorbidity in ST is unlikely to be a mere consequence of chronic disease and disfigurement. PMID- 12891095 TI - Screening for poststroke depression in a Hong Kong rehabilitation hospital: impact of different raters. PMID- 12891096 TI - Psychotropic drug use and expenditures among medicaid beneficiaries with and without other mental health or substance abuse services. PMID- 12891097 TI - Sexual relationship difficulties among borderline patients and axis II comparison subjects. PMID- 12891098 TI - Bright cyclic rearing protects albino mouse retina against acute light-induced apoptosis. AB - PURPOSE: Previous studies have shown that albino rats born and raised in bright cyclic light are protected from light-induced apoptosis. The present study was designed to determine if bright cyclic rearing provides protection against retinal degeneration caused by acute light exposure in albino mice. METHODS: BALB/c mice were born in dim cyclic light (5 lux, 12 h ON/OFF). At 1 week of age, half of the litters were moved into 400 lux cyclic light. At 5 weeks of age, mice raised in the dim or bright cyclic conditions were divided into two groups. One group was placed in constant light (3,000 lux for 72 h) and the other was maintained in its original cyclic light environment. Control and constant light stressed mice were dark-adapted for 24 and 48 h, respectively, after which their eyes were removed immediately for morphologic evaluation or preparation of rod outer segment (ROS) membranes. ROS lipids were extracted and fatty acid methyl esters were analyzed by gas-liquid chromatography. Eyes used for TUNEL (terminal deoxynucleotidyl transferase mediated dUTP nick end labeling) and DNA fragmentation assays were enucleated immediately after the 72 h light exposure. RESULTS: Measurement of outer nuclear layer (ONL) thickness indicated there was no difference in the number of viable photoreceptor cells in the dim-reared controls compared to bright-reared controls. Constant light exposure significantly reduced the ONL thickness in dim- and bright-reared groups, with the largest change occurring in the dim-reared mice. TUNEL assay showed no apoptotic photoreceptor cells in either control group; however, apoptotic nuclei could be detected in both exposed groups, with the largest number found in the dim-reared mice. After light exposure, DNA fragmentation was prominent in dim reared mice, but was not present in bright-reared animals. There was no significant difference in the fatty acid composition of ROS membranes in the dim- and bright-reared control mice. However, constant light exposure resulted in a greater loss of docosahexaenoic acid (22:6n-3) in the ROS of dim-reared animals. CONCLUSIONS: Mice raised in a bright cyclic light environment are protected against light-induced apoptosis. We suggest that the protection is due to the up regulation of cell survival pathways or the down-regulation of pathways that are vulnerable to acute cell stress. PMID- 12891100 TI - Open surgical and endovascular treatment of superior vena cava syndrome caused by nonmalignant disease. AB - OBJECTIVES: The purpose of this study was to evaluate the role of endovascular and open surgical reconstructions in patients with superior vena cava (SVC) syndrome caused by nonmalignant disease. METHODS: Clinical data from 32 consecutive patients who underwent endovascular or open surgical reconstruction of central veins because of symptomatic benign SVC syndrome between November 1983 and June 2001 were retrospectively reviewed. RESULTS: The study included 17 male and 15 female patients (mean age, 38 years; range, 5-69 years). Presenting symptoms were head fullness (n = 26), dyspnea or orthopnea (n = 23), headache (n = 17), or dizziness (n = 11); physical signs were head swelling (n = 31), chest wall collateral vessels (n = 29), facial cyanosis (n = 18), or arm swelling (n = 17). Etiologic factors included mediastinal fibrosis (n = 19), indwelling catheter (n = 8), idiopathic thrombosis (n = 4), or post-surgery (n = 1). Two patients were heterozygous for factor V Leiden; 1 patient had antithrombin III deficiency. Twenty-nine patients underwent surgical reconstruction with 31 bypass grafts: spiral saphenous vein (n = 20), superficial femoral vein (n = 4), human allograft (n = 1), or expanded polytetrafluoroethylene (ePTFE, n = 6). Eleven patients underwent percutaneous transluminal angioplasty or stenting; 3 primary and 8 secondary endovascular procedures were performed to treat graft stenosis (n = 7) or occlusion (n = 1). There were no early deaths. Five early graft failures in 3 ePTFE grafts and 2 bifurcated vein grafts (thrombosis, n = 4; stenosis, n = 1) were successfully treated with open surgical revision. Over a mean follow-up of 5.6 years (range, 0.4-16.6 years) in surgical patients, 17 additional secondary interventions were performed in 8 patients, 14 endovascular and 3 surgical. Primary, assisted primary, and secondary patency rates of surgical bypass grafts were 63%, 79%, and 85%, respectively, at 1 year, and 53%, 68%, and 80%, respectively, at 5 years. Graft patency was significantly higher in vein grafts compared with ePTFE grafts (P =.02). Mean follow-up after percutaneous transluminal angioplasty or stenting was 3.1 years (range, 1 day-11.7 years). Twelve secondary endovascular interventions were performed in 6 patients (primary group, 3 of 3; secondary group, 3 of 9 grafts in 8 patients) to maintain patency in 11 of 12 reconstructions. Mean follow-up in the entire patient cohort was 5.3 years (range, 0.4-16.6 years). In 79% of patients symptoms had resolved or were significantly improved at last follow-up. CONCLUSIONS: Surgical treatment of benign SVC syndrome is effective over the long term, with secondary endovascular interventions to maintain graft patency. Straight spiral saphenous vein graft remains the conduit of choice for surgical reconstruction, with results superior to those with bifurcated vein and ePTFE. Endovascular treatment is effective over the short term, with frequent need for repeat interventions. It does not adversely affect future open surgical reconstruction and may prove to be a reasonable primary intervention in selected patients. Patients who are not suitable for or who fail endovascular intervention merit open surgical reconstruction. PMID- 12891099 TI - Prospective randomized study of endovenous radiofrequency obliteration (closure procedure) versus ligation and stripping in a selected patient population (EVOLVeS Study). AB - PURPOSE: This study was designed as a prospective multicenter randomized comparison of procedure-related complications, patient recuperation, and quality of-life outcomes between patients undergoing vein stripping with high ligation and patients undergoing great saphenous vein (GSV) obliteration with temperature controlled radiofrequency ablation without adjunctive high ligation (Closure procedure). METHODS: Eighty-five patients (86 limbs) from five sites (France, 2; Austria, 1; United States, 2) were randomly allocated to undergo radiofrequency obliteration (RFO) or stripping and high ligation (S&L). Final analysis included data for 44 limbs in the RFO group and 36 limbs in the S&L group. Follow-up examinations were performed at 72 hours, 1 week, 3 weeks, and 4 months. All patients completed the CIVIQ2 quality-of-life (QOL) questionnaire and underwent clinical and ultrasound examinations at each follow-up visit. RESULTS: Immediate success on the day of treatment was reported for 95% (42 of 44) of limbs in the RFO group and 100% (36 of 36) of limbs in the S&L group. In seven RFO limbs (16.3%) a scan obtained 72 hours after the procedure showed flow in the proximal GSV. Five of these segments had reflux in the open segment. At 1 week two of these closed, and an additional segment closed at 3 weeks. In no cases did flow reappear after complete occlusion of the GSV. Time to return to normal activities was significantly less in the RFO group (mean, 1.15 days; 95% confidence interval [CI], 0.05-2.34) compared with the S&L group (mean, 3.89 days; CI, 2.67-5.12; P =.02). In the RFO group, 80.5% of patients returned to routine activities of daily living within 1 day, compared with 46.9% of patients in the S&L group (P <.01). Patients in the RFO group were able to return to work in 4.7 days (CI, 1.16-8.17), compared with 12.4 days (CI, 8.66-16.23) for the S&L group (P <.05). Analysis of the QOL surveys showed statistically significant differences in favor of the RFO group for global score and pain score during follow-up. The magnitude of the difference, however, progressively decreased between 1 week and 4 months. CONCLUSIONS: In the absence of significant complications, such as deep vein thrombosis and pulmonary embolism, severe neuritic sequelae, and skin burns, there are significant early advantages to endovascular obliteration of the GSV compared with conventional vein stripping. PMID- 12891101 TI - Validation of the new venous severity scoring system in varicose vein surgery. AB - OBJECTIVES: We performed this observational study to validate the three components of a new venous severity scoring (VSS) system, ie, venous clinical severity score (VCSS), venous segmental disease score (VSDS), and venous disability score (VDS), and to evaluate VCSS, VDS, and CEAP clinical class and score in quantifying outcome of varicose vein surgery. Patients and methods The study included 45 patients who underwent superficial venous surgery in 48 legs with primary varicose veins. Venous color duplex scanning, clinical examination, and a questionnaire were used preoperatively and at 6 weeks and 6 months postoperatively to assign VSS and CEAP clinical class and score. RESULTS: CEAP clinical score, VCSS, and VDS demonstrated a linear association with CEAP clinical class (P <.001, P <.001, P =.002, respectively). Good correlation among all severity scores was found, particularly between CEAP clinical score and VCSS (r = 0.94; P <.001). CEAP clinical score was also highly correlated with CEAP clinical class (r = 0.84; P <.001) and VDS (r = 0.70; P <.001). Similarly, VCSS correlated with CEAP clinical class (r = 0.83; P <.001) and also VDS (r = 0.72; P <.001). The anatomic severity marker VSDS demonstrated a weak correlation with clinical severity indicators VCSS (r = 0.29; P =.048) and VDS (r = 0.31; P =.03) but not with age, gender, or CEAP clinical class and score. Six months after surgery the median (interquartile range) percent change in VCSS (73%; range, 50% 100%) and CEAP clinical score (70%; range, 50%-100%) were both significantly greater (P <.001) than the corresponding change in CEAP clinical class (17%; range, 0%-50%). In legs with high VDS at baseline, median (interquartile range) percent change in VDS was 100% (range, 50%-100%), significantly greater (P <.001) than the corresponding change in CEAP clinical class (0%; range, 0%-17%). CONCLUSIONS: Venous severity scores are significantly higher in advanced venous disease, demonstrating correlation with anatomic extent. Both venous clinical severity scores, VCSS and CEAP clinical score, are equally sensitive and significantly better for measuring changes in response to superficial venous surgery than is the already in use CEAP clinical class. VDS demonstrated comparable and even better performance. Although the assignment of CEAP clinical class might be adequate for daily clinical purposes, venous severity scoring systems should be used in clinical studies to quantify venous outcome. PMID- 12891102 TI - A multicenter study of permanent hemodialysis access patency: beneficial effect of clipped vascular anastomotic technique. AB - OBJECTIVE: There is an urgent and compelling need to reduce the morbidity and expense of maintaining hemodialysis vascular access patency. This large, long term, retrospective, multicenter study, which compared access patency of autogenous arteriovenous fistulas (AVF) and synthetic bridge grafts (AVG) created with conventional sutures or nonpenetrating clips, was undertaken to resolve conflicting results from previous smaller studies. DESIGN: Patency data for 1385 vascular access anastomoses (clipped or sutured) was obtained from 17 hospitals and dialysis centers (Appendix). Five hundred eighteen AVF (242 clip, 276 suture) and 827 AVG (440 clip, 384 suture) were analyzed. Statistical comparisons were made with Kaplan-Meier survival analysis, log-rank test, two-sample t test, and X(2) test. The Cox proportional hazards model was used to confirm Kaplan-Meier analysis. RESULTS: Access patency (primary, secondary, overall, and intention to treat) was significantly improved in access anastomoses constructed with clips. In the intention-to-treat group, primary patency at 24 months was 0.54 for clipped AVF and 0.34 for sutured AVF, and was 0.36 for clipped AVG and 0.17 for sutured AVG. At 24 months, primary patency rate for AVF successfully used for dialysis was 0.67 for clips and 0.48 for sutures, and for AVG was 0.39 for clips and 0.19 for sutured constructs. Interventions necessary to maintain patency were significantly fewer in clipped anastomoses. CONCLUSION: Replacing conventional suture with clips significantly reduces morbidity associated with maintaining permanent hemodialysis vascular access. This beneficial effect may be due to the biologic superiority of interrupted, nonpenetrating vascular anastomoses. PMID- 12891103 TI - Timing and frequency of complications after carotid artery stenting: what is the optimal period of observation? AB - OBJECTIVE: Currently our standard of practice is that patients undergoing carotid artery stenting (CAS) may be safely discharged on the first day post-procedure. However, many patients are completely independent on the evening of procedure. Therefore we sought to establish the safety and feasibility of same-day discharge by assessing frequency and time of complications in the first 30 days after CAS. METHOD: Case records for 208 consecutive patients who had undergone CAS from October 1999 to October 2002 were retrospectively reviewed. Excluded were four cases in which combined CAS and carotid endarterectomy was performed to treat synchronous stenosis. Of the remaining 204 cases, involving 201 patients (three patients underwent staged bilateral CAS), 173 (84.8%) were symptomatic and 31 (15.2%) were asymptomatic. RESULT: Thirty-eight major events or death (inclusive of all neurologic events and any complications that required treatment) were recorded in the first 30 days after CAS, for an event rate of 18.6% per case. These were 7 (3.4%) major access site complications; 18 (8.8%) neurologic events, of which 10 (4.9%) were transient ischemic events and 8 (3.9%) were strokes (including minor, major, and fatal stroke); 8 (3.9%) cardiovascular complications; and five (2.5%) other events. Twenty (52.6%) events occurred in the first 6 hours after CAS, 2 (5.3%) between 6 and 12 hours, 3 (7.9%) between 12 and 24 hours, and 13 (34.2%) 24 hours to 30 days post procedure. Four (2.0%) deaths were recorded in this period, 1 secondary to stroke, 1 from a perforated myocardium from a temporary pacing wire, and 2 from unrelated causes. The 30-days stroke and death rate was 5.4% (11 of 204 patients). CONCLUSION: Time of complications suggests that outpatient performance of CAS is feasible and safe in selected patients. PMID- 12891104 TI - Carotid artery revascularization through a radiated field. AB - OBJECTIVE: Extracranial carotid stenosis is a complication of external head and neck irradiation. The safety and durability of carotid artery revascularization through a radiated field has been debated. We describe the immediate and long term results in a series of 27 consecutive patients who received treatment over 12 years. METHODS: From May 1990 to May 2002, 27 consecutive patients underwent 30 primary carotid artery revascularization procedures. All patients had received previous radiation therapy within a mean interval of 10 years (range, 1-26 years), with average radiation dose of 62 Gy (range, 50-70 Gy). Moderate to severe scarring of the skin or radiation fibrosis was present in three fourths of patients. Thirteen patients (48%) had undergone radical neck dissection, and 2 patients had a permanent tracheotomy. The indications for carotid surgery included high-grade (>70%) symptomatic stenosis in 18 patients (60%) and high grade asymptomatic stenosis in 12 patients (40%). General anesthesia with systematic shunting was used in 18 patients (60%), and regional anesthesia with selective shunting was used in 12 patients (40%). Operations included standard carotid endarterectomy (n = 20), with patch angioplasty (n = 12) or direct closure (n = 8); carotid interposition bypass grafting (n = 7); and subclavian to carotid bypass grafting (n = 3). Primary closure of the surgical wound was performed in all procedures without any special muscular or skin flaps. All patients were followed up for a mean of 40 months (range, 3-99 months). RESULTS: There was one (3.3%) perioperative death, from massive intracerebral hemorrhage; and 1 patient had a transient ischemic attack. In-hospital complications included neck hematoma in 2 patients, which required surgical drainage in 1 patient. There was neither delayed wound healing nor infection. Twelve patients died during follow-up, of causes not related to treatment. None of the surviving patients had further stroke, and all remained asymptomatic. Follow-up duplex scans showed asymptomatic recurrent stenosis greater than 60% in 3 patients, 2 of whom with stenosis greater than 80% underwent repeat operation. Risk for recurrent stenosis greater than 60% at 18 months was 16.6%. Recurrent stenosis occurred in 2 of these patients after saphenous vein bypass, and in 1 patient after endarterectomy with vein patch angioplasty. CONCLUSION: The clinical results and sustained freedom from symptoms and stroke over 40-month follow-up suggests that carotid revascularization through a radiated field is a safe and durable procedure in patients at high surgical risk, despite a marked incidence of recurrent stenosis. PMID- 12891105 TI - Postoperative infection associated with polyester patch angioplasty after carotid endarterectomy. AB - OBJECTIVES: Postoperative infection is one of the most dreaded complications associated with use of synthetic patches for carotid endarterectomy. Although polyester patches were used extensively for carotid patch angioplasty throughout the last decade, few reports detail cases of deep patch infection. We describe our experience with polyester patch infections after carotid endarterectomy. Patients and methods From January 1996 through December 2001 we treated polyester patch infections after carotid endarterectomy in 10 patients. RESULTS: The interval from primary carotid endarterectomy to presentation with infection ranged from 11 days to 30 months. All patients underwent repeat operation that involved tissue debridement, excision of the polyester patch, and either interposition grafting or patch angioplasty with autologous vein. No perioperative stroke or death occurred; however, 1 patient had transient hoarseness, and in 1 patient a pseudoaneurysm developed that required additional surgical repair with a saphenous vein interposition graft. All patients remain well and free of infection with follow-up as long as 56 months. CONCLUSIONS: Infection is a serious and rare complication of carotid patch angioplasty with polyester material. Nonetheless, it can be treated successfully with good results and acceptable morbidity with soft tissue debridement, prosthetic patch excision, and either patch angioplasty or interposition grafting with autologous vein. PMID- 12891106 TI - Neurologic outcome after penetrating extracranial arterial trauma. AB - PURPOSE: We undertook this study to determine factors that adversely affect outcome in patients with penetrating injury to the extracranial cerebral vasculature. Patients and methods Medical records were reviewed for all patients who had undergone surgical intervention to treat penetrating injury to the extracranial cerebral arteries between January 1989 and December 1999. Forensic autopsy findings were also reviewed for all patients who died as a result of their injury. RESULTS: One hundred fifty-one patients with injury to the brachiocephalic artery (n = 21), common carotid artery (n = 98), or internal carotid artery (n = 32) were identified. Overall mortality was 21.2%, and stroke rate in surviving patients was 15.1%. Twenty-five of 32 deaths (78.1%) were stroke-related. Brachiocephalic artery injury was associated with the highest mortality (38.1%), and survivor stroke rate was highest in patients with internal carotid injuries (22.7%). Hemodynamic instability at presentation led to both higher mortality (30.7%) and stroke rate (19.2%). Preoperative angiography did not influence mortality or stroke rate in hemodynamically stable patients. Procedural mortality associated with arterial ligation was 45% (9 of 20 patients), and no surviving patient experienced a change in pre-ligation neurologic state. Nine patients remained neurologically intact after ligation, and 2 patients with preoperative localized neurologic deficit were unchanged postoperatively. In 131 patients, mortality after arterial repair was 17.6%, and in 5 surviving patients (5.4%) an ischemic neurologic deficit developed. Twelve of 15 surviving patients (80%) with preoperative neurologic deficit who underwent arterial repair had improved neurologic status. Cerebral infarcts were confirmed at autopsy in 23 patients; 18 infarcts were ischemic (10, repair; 8, ligation), and 5 infarcts were hemorrhagic (all, repair). No factor was identified that was predictive of ischemic versus hemorrhagic infarction in patients undergoing repair. CONCLUSIONS: The presence of hypovolemic shock, internal carotid artery injury, complete vessel transection, and arterial ligation are associated with unfavorable outcome. Penetrating injury to the brachiocephalic, common carotid, or internal carotid artery should be repaired rather than ligated when technically possible. Subsequent ischemic or hemorrhagic cerebral infarction is unpredictable, but overall outcome is superior to that with ligation of the injured artery. PMID- 12891107 TI - Percutaneous arterial closure in peripheral vascular disease: a prospective randomized evaluation of the Perclose device. AB - BACKGROUND: Patients with peripheral vascular disease have been excluded from initial studies of percutaneous suture-mediated closure devices (SMCD) despite representing a significant proportion of those requiring endovascular intervention. We sought to determine whether these devices could be safely used in patients with peripheral vascular disease. METHODS: Patients were stratified into two groups and five subgroups on the basis of indication for arteriography, and they were prospectively randomized at the end of the procedure to receive either the SMCD or manual compression. Ankle-brachial index was determined and duplex ultrasound scanning of the accessed femoral artery was performed, before and after the procedure. Ultrasound data included peak systolic velocity, minimum intraluminal vessel diameter, and presence or absence of calcified plaque. Time to hemostasis, ambulation, and discharge were recorded, and major or minor complications were noted. RESULTS: Of 102 patients included in the study, 52 patients were randomized to receive the SMCD. There was no difference in ankle brachial index, minimum intraluminal vessel diameter, or peak systolic velocity in the accessed vessel after closure with SMCD or manual compression. Time to hemostasis, ambulation, and discharge was significantly less in the SMCD group (P =.001). Presence of calcified plaque was not associated with complications (P =.146). In the SMCD group, hemostasis was achieved with 49 of 52 devices (94.2%). There were six complications (5.9%), two of which were major and required operative intervention. All complications were hemorrhagic and not occlusive. There was no difference in overall complication rate between SMCD (7.7%) and manual compression (4.0%) groups (P =.678). No infection was noted in any of the 102 patients. CONCLUSIONS: Suture-mediated percutaneous arterial closure can be safely performed in patients with peripheral vascular disease, even in the presence of calcified plaque. This closure technique enables shorter time to hemostasis, ambulation, and hospital discharge. There are observed differences in minor, but not major, complication rates for MC versus percutaneous arterial closure in patients with peripheral vascular disease, but these differences did not achieve statistical significance in this small series. PMID- 12891108 TI - Iliac artery stenting versus surgical reconstruction for TASC (TransAtlantic Inter-Society Consensus) type B and type C iliac lesions. AB - OBJECTIVE: The TransAtlantic Inter-Society Consensus (TASC) document did not define the best treatment for moderately severe iliac artery lesions, ie, TASC type B and type C iliac lesions, because of insufficient solid evidence to make firm recommendations. The purpose of this study was to evaluate the influence of risk factors on outcome of iliac stenting and operative procedures used to treat TASC type B and type C lesions. METHODS: Over the 5 years from 1996 to 2001, 188 endovascular and direct aortoiliac surgical reconstruction procedures were performed in 87 women and 101 men with TASC type B and type C iliac lesions and chronic limb ischemia. The criteria prepared by the Ad Hoc Committee on Reporting Standards (Society for Vascular Surgery/International Society for Cardiovascular Surgery (SVS/ISCVS) were followed to define variables. Both univariate analysis (Kaplan-Meier method) and multivariate analysis (Cox proportional hazards model) were used to determine the association between variables, cumulative patency rate, limb salvage, and survival. RESULTS: Indications for revascularization were disabling claudication (73%), limb salvage (25%), and blue toe syndrome (2%). Patients in the surgery group (n = 52) had significantly higher primary patency rates compared with patients in the stent group (n = 136) at univariate analysis (Kaplan-Meier method, log-rank test; P =.015). Primary patency rates at 1, 3, and 5 years were 85%, 72%, and 64% after iliac stenting, and 89%, 86%, and 86% after surgical reconstruction, respectively. Univariate and multivariate Cox regression analysis enabled identification of poor runoff (ie, runoff score >5 for unilateral procedures or >2.5 for bilateral outflow procedures; relative risk, 2.5; 95% confidence interval [CI], 1.4-4.2; P =.001) as the only independent predictor of decreased primary patency in all patients. However, stratified analysis including only patients with poor runoff revealed that patients undergoing iliac stenting had significantly lower primary patency rates compared with those undergoing surgical reconstruction (Kaplan-Meier method, log-rank test; P =.05). External iliac artery disease and female gender were also identified as independent predictors of decreased primary stent patency. CONCLUSIONS: Poor infrainguinal runoff is the main risk factor for decreased primary patency after surgical reconstruction and iliac stenting to treat TASC type B and type C iliac lesions. However, primary patency is less affected by poor runoff in patients undergoing surgical procedures. The presence of poor runoff, external iliac artery disease, and female gender are independent predictors of poor outcome after iliac stenting, and therefore these risk factors should determine the need for surgical reconstruction. PMID- 12891109 TI - Outcome of revascularization procedures for peripheral arterial occlusive disease in Ontario between 1991 and 1998: a population-based study. AB - PURPOSE: We describe the outcome of revascularization procedures used to treat peripheral arterial occlusive disease (PAOD), using population-based administrative data. METHODS: A retrospective population-based cohort study utilizing administrative databases in Ontario, Canada, was conducted for fiscal years 1991 to 1998 to identify patients who underwent arterial bypass surgery and percutaneous transluminal angioplasty to treat PAOD. The Kaplan-Meier method was used to calculate cumulative survival rate and amputation-free survival rate. To analyze factors that affect these rates, multivariate analysis was performed with Cox proportional hazard models. RESULTS: Over the study period 15,824 patients underwent bypass operations and 11,548 underwent angioplasty. For patients who underwent bypass surgery, 5-year cumulative survival rate was 61.5% and major amputation-free survival rate was 83.4%, compared with 69% and 92.2%, respectively, for patients who underwent angioplasty. Male sex, older age, diabetes, and heart disease were associated with increased risk for death after revascularization procedures. Increased risk for major amputation after revascularization procedures was associated with male sex, older age, and diabetes, whereas hypertension was linked to decreased risk. CONCLUSION: To evaluate the long-term outcome of revascularization procedures for PAOD at the population level, survival and major amputation-free survival rates should be used, because they provide more clinically accepted estimates compared with the correlation between utilization rates for revascularization and amputation procedures, which have been used to describe outcome in previously published reports in the literature. PMID- 12891110 TI - Use of interventional procedures for peripheral arterial occlusive disease in Ontario between 1991 and 1998: a population-based study. AB - PURPOSE: Although peripheral arterial occlusive disease (PAOD) is a public health issue in the elderly population, limited population-based data are available on use of interventional procedures in Canada. We describe trends in use of interventional procedures to treat PAOD in Ontario over the past decade. METHODS: A retrospective population-based cohort study was conducted for fiscal years 1991 to 1998 with Ontario administrative databases to identify all arterial bypass surgeries, angioplasty procedures, and amputations performed. RESULTS: A total of 19,332 bypass operations, 16,334 angioplasty procedures, and 17,534 amputations were identified. Population-based rates showed that angioplasty use peaked at about 110 per 100,000 at age 65 to 74 years, arterial bypass surgery use peaked at 129 per 100,000 at age 75 to 84 years, and amputation use peaked at 138 per 100,000 at age 85 years or older. All types of interventional procedures to treat PAOD were performed more frequently in men than in women. Age-adjusted and sex adjusted rate of arterial bypass surgery decreased significantly, from 77 to 61 per 100,000 population aged 45 years or older (P =.0002, linear regression analysis), whereas rate for PTA increased significantly, from 59 to 75 per 100,000 population aged 45 years or older (P =.0005). The overall major amputation rate declined slightly over the study period, influenced by the decreased rate in patients aged 85 years or older. The revascularization rate in patients aged 85 years or older increased (P =.055). CONCLUSION: Reduced use of arterial bypass surgery and increased use of angioplasty procedures has occurred over the past decade and may reflect a change in the practice pattern of vascular surgeons in Ontario, who have become more conservative in treating localized disease and reserve surgical interventions for more severe forms of PAOD. The slight reduction in overall major amputation rate, driven by decreased rate in patients aged 85 years or older, may reflect a trend toward a more aggressive revascularization approach in this age group. PMID- 12891111 TI - Evaluating effects of method of administration on Walking Impairment Questionnaire. AB - BACKGROUND: Intermittent claudication resulting from peripheral arterial disease (PAD) can substantially impair walking function. The Walking Impairment Questionnaire (WIQ) assesses patient self-reported difficulty in walking. Currently this questionnaire is validated for interviewer administration only. Since this can be burdensome in a large clinical trial, we examined the effects of alternative methods of administration on patient responses on the WIQ. METHODS: The WIQ, which consists of four subscales (pain severity, distance, speed, stairs), was modified to be self-administered or interviewer-administered by telephone. Patients with PAD were recruited from two sites and randomized into two groups: in group 1 the WIQ was self-administered, then telephone administered; in group 2 the WIQ was telephone-administered, then self administered. The two administrations occurred 4 to 7 days apart. Additional measures (SF-36, EQ-5D, and PAD symptom scale) and clinical data were included to further assess the validity of the WIQ and symptoms in patients with claudication. Telephone interviews were conducted by trained interviewers using standardized scripts. Two-week test-retest reliability was assessed for both the self-administered WIQ (group 1) and the telephone-administered WIQ (group 2). RESULTS: Sixty patients were recruited at two sites (n = 30 per group). Seventy eight percent were men; mean patient age was 67.1 years; and 83% of patients were white. Mean duration of PAD symptoms was 6.8 years. No significant differences were observed in WIQ subscale scores between self administration and telephone administration. No interaction effects between order and method of administration were detected. Cronbach alpha for distance, speed, and stair-climbing subscales ranged from 0.82 to 0.94. Correlations among WIQ subscales and the symptom scale were good (r = -0.34 to -0.57). Correlations of WIQ subscales with physical health subscales of the SF-36 (r = 0.24-0.59) were higher than for mental health related subscales (r = 0.08-0.26). CONCLUSIONS: The modified WIQ demonstrated good reliability and validity with both methods of administration. These results suggest that the self-administered and telephone-administered versions of the WIQ can be used reliably and efficiently in clinical trials. PMID- 12891112 TI - Biologic degeneration of vein grafts after thrombotic occlusion: thrombectomy within 3 days results in better indices of viability. AB - OBJECTIVES: To clarify the mechanism for poor patency of vein grafts after thrombectomy and the time limit for successful salvage operation, we investigated the time course of biologic degenerative changes in thrombosed vein grafts. Materials and methods The right femoral artery was replaced with a femoral vein graft in 25 mongrel dogs. After 3 months, grafts were explanted in 5 dogs (control grafts), and the remaining 20 dogs underwent femoral artery ligation to create a thrombosed graft. Of the 20 grafts, 5 were explanted at 3 days after ligation (group I-3) and 5 were explanted at 5 days after ligation (group I-5). Of the remaining 10 grafts, 5 underwent thrombectomy at 3 days after ligation (group II-3) and 5 underwent thrombectomy at 5 days after ligation, and were reimplanted into the left femoral artery, then explanted 28 days after reimplantation. The grafts were assessed with immunohistochemistry and prostaglandin (PG) I(2) assay (6-keto-PGI(1alpha)). RESULTS: Of the 25 grafts, occlusion recurred in 3 in group II-5 within 28 days after reimplantation. There were significant differences between group I-5 and group I-3 or control grafts for percentage of areas positive for alpha-actin, total number of cells per field, and proliferating cell nuclear antigen (PCNA)-positive cells in layer of thickened intima and atrophied media (I/M), and for total cell and PCNA- positive cell numbers per field in the adventitia. Mean 6-ketoPGF(1alpha) was 40 +/- 14.1 pg/mg/min in control dogs, 84 +/- 18.9 pg/mg/min in group I-3, and 15.4 +/- 7.7 pg/mg/min in group I-5, demonstrating a significant reduction in group I-5 (P =.009). CONCLUSION: Graft wall cell viability and PGI(2) production in thrombosed vein grafts are well preserved for up to 3 days. Therefore graft salvage operations no later than 3 days after thrombotic occlusion may provide acceptable long-term patency of salvaged grafts. PMID- 12891113 TI - Safety of gadolinium contrast angiography in patients with chronic renal insufficiency. AB - OBJECTIVE: To prevent iodinated contrast medium-induced nephrotoxicity, gadolinium has been used increasingly for magnetic resonance angiography (MRA) or conventional digital subtraction angiography (DSA) to visualize arterial anatomy in patients undergoing vascular surgery who are considered at high risk because of chronic renal insufficiency. We assessed the safety of gadolinium-based contrast medium as a substitute for iodinated contrast medium-enhanced examinations. We determined the incidence of gadolinium-induced nephrotoxicity in a clinical setting and searched for contributing risk factors. Patients and methods In a single-center retrospective study from December 1999 to January 2001, 218 inpatients underwent MRA and 42 inpatients underwent DSA, with gadolinium as the sole contrast agent. Patient comorbid conditions, indications for vascular imaging, contrast dose, urine output, baseline and post-procedure serum creatinine concentration (SCr), and outcome were recorded for all patients in whom gadolinium-induced renal failure developed. RESULTS: Of 260 patients who received gadolinium-based contrast agents, at a dose of 0.25 mmol/kg or more, 195 patients (75%) had pre-test baseline chronic renal insufficiency. In 7 of 195 patients (3.5%) acute renal failure developed after gadolinium-based contrast medium administration, for MRA (n = 153) in 3 patients (1.9%) and DSA (n = 42) in 4 patients (9.5%). Average baseline SCr in the 195 patients with chronic renal insufficiency was 38.2 +/- 1.6 mL/min/1.73 m(2), and in the 7 patients in whom acute renal failure developed, baseline SCr was 32.5 +/- 7.8 mL/min/1.73 m(2) (P =.33). Respective intravenous and intra-arterial gadolinium doses in these 7 patients ranged from 0.31 to 0.41 mmol/kg for MRA and 0.27 to 0.42 mmol/kg for DSA. Acute renal failure did not develop in any of 65 patients with normal baseline SCr. CONCLUSION: Despite reports of negligible nephrotoxicity, rarely gadolinium-based contrast agents can cause acute renal failure in patients with underlying chronic renal insufficiency. Estimation of creatinine clearance alone does not enable prediction of which patients are likely to have acute renal failure. Patients at high-risk should be identified, and prophylactic measures should be taken to reduce the risk for nephrotoxicity. PMID- 12891115 TI - Aortoiliac morphologic correlations in aneurysms undergoing endovascular repair. AB - PURPOSE: The feasibility of endovascular aneurysm repair depends on morphologic characteristics of the aortoiliac segment. Knowledge of such characteristics is relevant to safe deployment of a particular device in a single patient and to development of new devices for use in patients with a broader spectrum of anatomic variations. METHODS: We evaluated findings on computed tomography scans for 277 patients being considered for endovascular aneurysm repair. Aortic neck length and angulation estimates were generated with three-dimensional trigonometry. Specific centerline points were recorded, corresponding to the aorta at the celiac axis, lowest renal artery, cranial aspect of the aneurysm sac, aortic terminus, right hypogastric artery origin, and left hypogastric origin. Aortic neck thrombus and calcium content were recorded, and neck conicity was calculated in degrees. Statistical analysis was performed with the Spearman rank correlation. Data are expressed as median and interquartile range. RESULTS: Median diameter of the aneurysms was 52 mm (interquartile range, 48-59 mm) in minor axis and 56 mm (interquartile range, 51-64 mm) in major axis, and median length was 88 mm (interquartile range, 74-103 mm). Median proximal aortic neck diameter was 26 mm (interquartile range, 22-29 mm), and median neck length was 30 mm (interquartile range, 18-45 mm). The common iliac arteries were similar in diameter (right artery, 16 mm [interquartile range, 13-20 mm]; left artery, 15 mm [interquartile range, 11-18 mm]) and length (right, 59 mm [interquartile range, 50-69 mm]; left, 60 mm [interquartile range, 49-70 mm]). Median angulation of the infrarenal aortic neck was 40 degrees (interquartile range, 29-51 degrees), and median angulation of the suprarenal segment was 45 degrees (interquartile range, 36-57 degrees). By gender, sac diameter, proximal neck diameter, and iliac artery diameter were significantly larger in men. Significant linear associations were identified between sac diameter and sac length, neck angulation, and iliac artery diameter. As the length of the aneurysm sac increased the proximal aortic neck length decreased. Conversely, as the sac length decreased sac eccentricity increased. Mural thrombus content within the neck increased with increasing neck diameter. CONCLUSIONS: There is considerable variability in aortoiliac morphologic parameters. Significant associations were found between various morphologic variables, links that are presumably related to a shared pathogenesis for aberration in aortoiliac diameter, length, and angulation. Ultimately this information can be used to develop new endovascular devices with broader applicability and improved long-term results. PMID- 12891114 TI - Ruptured thoracoabdominal aortic aneurysm treatment in the United States: 1988 to 1998. AB - OBJECTIVE: Thoracoabdominal aortic aneurysm (TAAA) rupture usually results in death. The outcome remains poor for patients who reach the operating room. The objective of this investigation was to define contemporary surgical experience with ruptured TAAA in the United States. Patients and methods Clinical data derived from the Nationwide Inpatient Sample on patients who underwent repair of a ruptured TAAA from 1988 to 1998 were analyzed. Age, sex, race, nature of admission, comorbid conditions, and provider volume were abstracted from the database. In-hospital mortality, postoperative complications, and length of stay were the principal outcome measures. RESULTS: Three hundred twenty-one patients were identified for the study. Mean age was 71.5 years; men outnumbered women (63% vs 37%). Crude overall surgical mortality was 53.8% and did not improve over time. Operative mortality was most likely (51%) to occur within the first 24 hours postoperatively. Median length of stay for surviving patients was 16 days. Renal failure (28%) and cardiac complications (18%) were the most common complications. In a logistic regression model, age greater than 77 years was predictive of death (odds ratio [OR], 2.5; P =.005), and nonwhite race appeared protective (OR, 0.53; P =.013). CONCLUSIONS: Mortality after surgical treatment of ruptured TAAA is high. Surviving patients experience many postoperative complications and have lengthy hospital stays. Given the lack of significant improvement in contemporary surgical practice, new techniques of repair deserve the attention of clinicians. PMID- 12891116 TI - Smokers' relative risk for aortic aneurysm compared with other smoking-related diseases: a systematic review. AB - OBJECTIVE: Aortic aneurysm has traditionally been considered a manifestation of atherosclerosis, but recent evidence suggests an independent pathogenesis, possibly similar to that of chronic obstructive pulmonary disease (COPD). Further insight into the pathogenesis of aortic aneurysm might be obtained by comparing its association with smoking with that of other smoking-related diseases. STUDY DESIGN: We conducted a systematic review of studies providing relative risk associated with smoking for both aortic aneurysm and other smoking-related diseases. RESULTS: We identified 10 eligible studies, which included more than 3 million subjects. The events reported in 9 studies were death from target diseases; the tenth study reported new diagnoses. Relative risk for aortic aneurysm-related events in current smokers was generally 3 to 6, compared with 1 to 2 for coronary artery disease or cerebrovascular disease and 5 to 12 for COPD. For each category of smoking in each study, relative risk associated with smoking was substantially greater for aortic aneurysm than for coronary artery disease or cerebrovascular disease. Our pooled estimates indicate that, in men, the association of ever smoking with aortic aneurysm is 2.5 times greater than the association of ever smoking with coronary artery disease (95% confidence interval [CI], 2.2, 2.8) and 3.5 times greater than the association of ever smoking with cerebrovascular disease (95% CI, 2.4, 5.3), but only 0.56 as great as the association of ever smoking with COPD (95% CI, 0.36, 0.86). CONCLUSIONS: The difference in magnitude of these associations with smoking is consistent with a non-atherosclerotic cause for aortic aneurysm and/or a stronger effect of smoking on vascular disease in the peripheral arteries. PMID- 12891117 TI - Factors that predict prolonged length of stay after aortic surgery. AB - OBJECTIVES: In this era of managed health care, third-party payers insist that surgeons minimize hospital stay even after major operations such as aortic surgery. We attempted to identify risk factors that predict prolonged hospital length of stay (LOS) so that realistic expectations can be established for these patients who frequently are at high-risk. METHODS: In 1994 a clinical pathway for aortic surgery was implemented at our hospital. Between January 1, 1994, and December 31, 2000, data including identifiable risk factors and LOS were reviewed for 240 patients who underwent elective infrarenal aortic surgery to treat aneurysmal (n = 179) or occlusive (n = 61) disease. Risk factors were analyzed to determine their effect on LOS. Data for patients who underwent endovascular, emergency, or concomitant cardiac surgery were excluded from analysis. RESULTS: In-hospital mortality was 0.4% (1 of 240 patients), and morbidity was 18% (44 of 240 patients). Mean LOS was 8.2 +/- 5.7 days for all patients, 6.9 +/- 2.9 days for those without complications, and 13.8 +/- 6.7 days for patients with complications (P <.0001). Factors that predicted prolonged LOS (Kaplan-Meier method) included age older than 75 years (P =.0004), chronic obstructive pulmonary disease (COPD; P =.0351), intraoperative blood loss more than 500 mL (P =.0006), duration of surgery more than 5 hours (P <.0001), wound infection (P =.0311), and postoperative complications overall (P <.0001). Remaining factors associated with prolonged LOS (Cox regression analysis) included age older than 75 years (P =.0050), COPD (P =.0445), and complications overall (P =.0094). CONCLUSION: The only identifiable preoperative risk factors that correlated with increasing LOS after elective infrarenal aortic surgery (multivariate analysis) were increasing age and COPD. Third-party payers should allow longer hospitalization for patients older than 75 years and for patients with significant pulmonary disease. PMID- 12891118 TI - Videodensitometric blood flow analysis of abdominal aortic aneurysm and intravascular coagulation. AB - OBJECTIVE: We investigated the relation between flow pattern in abdominal aortic aneurysm (AAA) and intravascular coagulopathy characterized by increased fibrin degradation product d-dimer (FDP-DD) or thrombin-antithrombin complex (TAT). Materials and methods The ratio of AAA maximum endoluminal diameter (diameter of flow channel) (n = 23) to diameter of the aorta between the superior mesenteric and renal arteries (R ratio) was measured with three-dimensional computed tomography angiography. Digital subtraction angiography was performed with 20 mL (10 mL/s) of contrast agent injected from the suprarenal portion of the abdominal aorta. The duration between the time when average gray scale in the AAA reached maximum and average region of interest gray scale decreased to half-maximum (bolus transit time in AAA [BTT(AAA)]) was calculated. RESULTS: Single correlation coefficient with statistic significance was detected between R ratio and BTT(AAA) (BTT(AAA) = 2.54 x R ratio + 3.65; r(2) =.30; P =.042). Among the three-dimensional morphologic and videodensitometric variants, BTT(AAA) was the most determinant factor associated with FDP-DD (FDP-DD = - 8.647 + 2.029 x BTT(AAA); r(2) =.448; P =.005). The most efficient predictors for TAT were maximum AAA endoluminal diameter (R(endomax)) and BTT(AAA) (TAT = - 14.007 + 2.102 x BTT(AAA) + 0.296 x R(endomax), r(2) =.360; P =.0069). CONCLUSIONS: Our findings suggest a close link between abnormal flow pattern in AAA and activation of the coagulation-fibrinolysis system. Videodensitometric blood flow analysis can be useful in investigation of the pathophysiology of phenomena related to abnormal flow field in AAA. PMID- 12891119 TI - Atrophy of the abdominal wall muscles after extraperitoneal approach to the aorta. AB - OBJECTIVE: We retrospectively assessed computed tomography (CT) scans to determine degree of anterolateral abdominal muscle atrophy in patients who underwent infrarenal aortic repair with 2 kinds of incisions for the extraperitoneal approach. METHODS: CT scans obtained before surgery and final scans obtained 2 to 100 months after surgery were assessed in 12 patients with paramedian incision (PM group) and 27 patients with flank incision (F group) who could be followed up at our hospital. We considered muscle thickness before surgery on the incision side to be 100% thickness (baseline value), and we calculated, by measuring the incision side after surgery, the corrected percent thickness (CPT%), which represents percentage of remaining muscle thickness that has escaped incision-induced atrophy. CT scans obtained at the level of the third (L3) and fifth (L5) lumbar vertebrae and the center of the sacrum (S) were selected for CPT% measurement. RESULTS: Duration from surgery to final CT scan was 2 to 65 months (mean +/- SD, 34.33 +/- 21.38 months) in PM group and 3 to 96 months (27.85 +/- 20.74 months) in F group. In PM group, mean CPT% values of the rectus abdominis muscle were 55.83 +/- 21.65% at L3, 35.50 +/- 10.79% at L5, and 31.92 +/- 11.00% at S; these values were statistically much smaller than baseline (P <.01). Mean CPT% values of the lateral abdominal muscles were not statistically different from baseline. In F group, mean CPT% values of the rectus abdominis muscle were 82.19 +/- 23.15% at L5 and 64.41 +/- 31.34% at S; these values were statistically smaller than baseline (P <.01). Mean CPT% values of the lateral abdominal muscles were 87.59 +/- 22.30% at L3 and 84.59 +/- 26.90% at L5; these values were statistically smaller than baseline (P <.05). CONCLUSIONS: Paramedian incision induced severe rectus abdominis muscle atrophy. Although flank incision induced various degrees of atrophy in both muscles, some patients had no muscle atrophy. These data indicate that further anatomic investigation into the relation between flank incision and abdominal wall innervation may contribute to prevention of muscle atrophy after flank incision. PMID- 12891120 TI - Prostaglandin E2 receptors in abdominal aortic aneurysm and human aortic smooth muscle cells. AB - BACKGROUND: Prostaglandin (PG) E(2) (PGE(2)) appears to have a role in stimulating production of interleukin-6 (IL-6) and apoptosis of smooth muscle cells in diseased aortic tissue. These actions are mediated by cellular receptors for PGE(2) EP receptors. OBJECTIVE: This study was undertaken to identify EP receptors associated with production of IL-6 by aortic explants. METHODS: Biopsy specimens of abdominal aortic aneurysm were used for explant culture and preparation of messenger RNA. The presence of EP1, EP2, EP3, and EP4 receptors in tissue and cells was investigated with reverse-transcriptase polymerase chain reaction. IL-6 and cyclic adenosine monophosphate were measured with an enzyme linked immunosorbent assay. RESULTS: PGE(2) or 11-deoxy-PGE(1) (EP 2/3/4 agonist) reversed partially the indomethacin suppression of IL-6 secretion from explant cultures, whereas butaprost (EP2 receptor agonist) and sulprostone (EP 1/3 receptor agonist) had no effect. Aortic biopsy specimens expressed EP2, EP3-III, and EP4 receptors. Aortic smooth muscle cells expressed EP2 receptor and four variants of EP3 receptor, ie, EP3-Ib, EP3-II, EP3-III, and EP3-IV, but PGE(2) did not stimulate secretion of IL-6. In contrast, PGE(2) or 11-deoxy-PGE(1) stimulated secretion of IL-6 from aortic macrophages. CONCLUSIONS: In aortic explants, PGE(2) stimulates IL-6 secretion by activation of EP4 receptors, present in macrophages. PMID- 12891121 TI - Inducible nitric oxide synthase is present in human abdominal aortic aneurysm and promotes oxidative vascular injury. AB - OBJECTIVE: Nitric oxide (NO), catalyzed by inducible NO synthase (iNOS), may be important in the pathophysiologic characteristics of many vascular diseases. Although there is indirect evidence to support the presence of iNOS in abdominal aortic aneurysm (AAA) in human beings, no definitive study has confirm this finding. The present study was designed to assess expression of iNOS in AAA in human beings. Furthermore, the activity of iNOS and the oxidative vascular injury initiated by iNOS were assessed with detection of nitrotyrosine, which is a marker indicative of formation and activity of the NO-derived oxidant peroxynitrite. METHODS: We studied 25 patients with AAA and 10 patients with normal abdominal aortas. In situ hybridization and immunohistochemistry were used in tissue sections to localize iNOS messenger RNA (mRNA) and protein. Double staining with a combination of in situ hybridization and immunohistochemistry was used to simultaneously demonstrate iNOS mRNA expression and its cellular localization. The presence of peroxynitrite was indirectly assessed with immunostaining with anti-nitrotyrosine antibodies. RESULTS: In situ hybridization and immunohistochemistry confirmed the presence of iNOS in media and adventitia of AAA in all 25 patients. Specific cell markers identified iNOS mRNA-positive cells mainly as T and B lymphocytes, macrophages, and smooth muscle cells. Positive immunostaining for nitrotyrosine was present in macrophages and smooth muscle cells. Normal abdominal aorta demonstrated virtually no iNOS or nitrotyrosine expression. CONCLUSION: Stimulated expression of iNOS is associated with degeneration of AAA in human beings, and the activity of this enzyme under such conditions preferentially promotes formation and activity of peroxynitrite and further contributes to oxidative tissue and cellular injury in AAA. This may be important in the pathogenesis of AAA. PMID- 12891122 TI - Graft-preserving treatment for vascular graft infected with Staphylococcus aureus with antibiotic-releasing porous apatite ceramic in the rabbit. AB - OBJECTIVE: This study was undertaken to investigate whether infection of a vascular graft with Staphylococcus aureus can be treated in situ by applying antibiotic-loaded porous apatite ceramic, in a rabbit model. METHODS: Teicoplanin (TEIC) was loaded onto a beta-tricalcium phosphate (TCP) block, a type of porous apatite ceramic. The activity of TEIC released from the antibiotic-loaded TCP block was examined in vivo. A vascular graft was patched onto the abdominal aorta in 24 rabbits, and S aureus was applied directly on it. Seven days postoperatively, each rabbit underwent repeat laparotomy, and retroperitoneal abscess around the prosthetic vascular patch was debrided. Animals were divided into four groups of 6 rabbits each. In group 1 only debridement was carried out. In groups 2 and 3, solution containing 40 or 60 mg of TEIC, respectively, was applied to the prosthetic vascular patch. In group 4, an antibiotic-loaded TCP block (63 +/- 6.6 mg of TEIC) was placed around the graft. Three weeks after the second operation, the graft, the tissue around it, and arterial blood were collected and cultured. RESULTS: TEIC activity was maintained for 28 days in vivo. In group 1, bacterial cultures of the prosthetic vascular graft and the tissue around it were positive in 5 animals and negative in 1 animal (infection rate, 83%). In both groups 2 and 3, cultures were positive in 3 animals and negative in 3 animals (infection rate, 50%). In group 4, cultures were negative in all animals (infection rate, 0%). Blood cultures were negative in all animals. Infection rate in group 4 was significantly lower than that in group 1 (P =.03), and was also lower than that in groups 2 and 3, but the difference was not significant. CONCLUSIONS: Use of slow-release antibiotic loaded onto a TCP block, along with debridement, may control infection in vascular grafts in situ, averting the necessity to remove the graft. PMID- 12891123 TI - Endothelial dysfunction in peripheral arterial disease is related to increase in plasma markers of inflammation and severity of peripheral circulatory impairment but not to classic risk factors and atherosclerotic burden. AB - OBJECTIVE: We undertook this study to evaluate in patients with peripheral arterial disease (PAD) the relationship of endothelial dysfunction, which is directly related to progression and clinical complications of atherosclerosis, with variables including classic risk factors, inflammation, severity of peripheral circulatory impairment, and atherosclerotic burden. METHODS: This cross-sectional study included outpatients seen in an academic angiologic unit. Eighty-eight consecutive patients with PAD (ankle/brachial index [ABI] < 0.90) were studied. The control group consisted of 30 age-matched and sex-matched healthy subjects. Main outcome measures were endothelial function in the form of brachial artery flow-mediated dilation (FMD), plasma levels of C-reactive protein (CRP) and fibrinogen, severity of PAD according to ABI, and atherosclerotic burden, ie, atherosclerosis in one leg or in two or more other sites. RESULTS: Compared with patients with FMD greater than 6.2% (ie, 5th percentile of FMD in control subjects), patients with FMD less than 6.2% had a similar prevalence of classic risk factors but higher median levels of CRP (1.6 vs 6.0 mg/L; P <.01) and fibrinogen (200 vs 374 mg/dL; P <.01). The two inflammatory markers were negatively correlated with FMD (P <.01). ABI was higher in patients with FMD greater than 6.2% than in those with worse endothelial function (0.72 +/- 0.15 vs 0.62 +/- 16; P <.01); there was no difference with respect to atherosclerotic burden. Multivariate analysis showed that the association of CRP, fibrinogen, and ABI with FMD less than 6.2% was unrelated to classic risk factors. In a second model, which included CRP, fibrinogen, and ABI, all three variables were independently related to FMD less than 6.2%. CONCLUSION: Inflammation and severity of circulatory impairment are implicated in the pathophysiology of dysfunctional endothelium in PAD. PMID- 12891124 TI - Proximal aortic stent-graft displacement with type I endoleak due to Heimlich maneuver. AB - The Heimlich maneuver has saved countless lives by relieving foreign body obstruction from the respiratory tract. Complications related to this life-saving technique, particularly involving the abdominal aorta, are extremely rare. We report the case of a patient who underwent successful endovascular abdominal aortic aneurysm (AAA) repair, with AAA reduction at postoperative surveillance. Endograft displacement after performance of the Heimlich maneuver resulted in a proximal type I endoleak. This case underscores both the potential for aortic trauma from the Heimlich maneuver and the risk for clinical failure as a result of abdominal compression after successful endovascular AAA repair. PMID- 12891125 TI - Endovascular stent-graft repair for penetrating atherosclerotic ulcer in the infrarenal abdominal aorta. AB - PURPOSE: Penetrating atherosclerotic ulcer (PAU) is an ulceration of an atherosclerotic plaque penetrating through the intima, which may lead to intramural hematoma, aneurysm formation, or rupture. This disease is predominantly found in the thoracic aorta and is uncommon in the infrarenal aorta. The effectiveness of endovascular repair of PAU in the infrarenal aorta was retrospectively investigated. METHODS: From 1999 to 2002, PAU was diagnosed with computed tomography and magnetic resonance imaging in the abdominal aorta in four patients. All patients were men; their average age was 78 years. All four patients had hypertension, and two patients had concomitant coronary artery disease. Three patients had abdominal pain or lumbago. RESULTS: All patients underwent endovascular grafting with a Gianturco Z-stent covered with thin-wall woven Dacron graft. Indications for endovascular intervention were aneurysm formation with or without intramural hematoma in two patients and contained rupture with extraaortic hematoma in two patients. The postoperative course was uneventful in all cases, and no endoleak or aneurysm expansion was recognized during follow-up (4-32 months; average, 14 months). CONCLUSIONS: Infrarenal aortic lesions caused by PAU were generally localized, and endovascular grafting appears to be a feasible alternative to surgical repair. PMID- 12891126 TI - New approach to preoperative vascular exclusion for carotid body tumor. AB - We report a new approach to preoperative vascular exclusion of a carotid body tumor. Before surgery, covered stents were placed in the external carotid artery, resulting in vascular exclusion of the tumor. Subsequent surgical excision was uneventful, with operative blood loss less than 200 mL and no neurologic complications postoperatively. This technique deserves further consideration as a reasonable alternative to conventional embolization. PMID- 12891127 TI - Spontaneous arterial thrombosis in a patient with human immunodeficiency virus infection: successful treatment with pharmacomechanical thrombectomy. AB - Patients with human immunodeficiency virus (HIV) have various coagulation abnormalities as well as increased risk for development of clinical thrombosis and subsequent embolic events. We report acute lower leg ischemia caused by spontaneous atheroembolism with no identifiable source in a young patient with HIV infection. Treatment included percutaneous mechanical thrombectomy and thrombolysis, which reversed the arterial ischemia. Physicians should be aware of thromboembolic disease as a possible complication of HIV. PMID- 12891128 TI - Successful stent-graft exclusion of a bovine patch-related carotid artery pseudoaneurysm. PMID- 12891129 TI - Salvage of a difficult situation: method for conversion of failed endograft. AB - Complications of endovascular aneurysm repair can be difficult to manage. One of the more difficult situations is conversion to open surgery because of a failed endograft. We describe a technique for conversion that allows the proximal attachment system to remain intact. It may also enable infrarenal clamping of the aorta during the operation. The anastomosis is performed by incorporating the proximal attachment system of the endograft. This technique simplifies both vascular control of the aorta and the necessary reconstruction during creation of the proximal anastomosis. We believe this technique has important advantages when conversion of a failed endograft is required. PMID- 12891130 TI - Technique to decrease lower extremity and pelvic ischemia during thoracoabdominal aortic aneurysm repair. PMID- 12891131 TI - Current modifications to totally laparoscopic "apron technique". AB - Since our original description in 1997 of a totally laparoscopic technique for treatment of aortoiliac disease, this type of minimally invasive procedure has been used both in the United States and abroad. We describe improvements that should make this technique more easily reproducible. This modified procedure was offered to six patients, one of whom received a tube graft for treatment of aneurysm disease. PMID- 12891132 TI - Ethics of institutional marketing: role of physicians. PMID- 12891142 TI - Web training: start early. PMID- 12891143 TI - Healthy eating and exercise in the rural midwest. PMID- 12891144 TI - Scholarship and the CDR recertification (Professional Development Portfolio) process. PMID- 12891145 TI - The Lenna Francis Cooper Memorial Lecture--The future of clinical dietetics: evidence, outcomes, and reimbursement. PMID- 12891146 TI - The association between vitamin C and vitamin E supplement use before hematopoietic stem cell transplant and outcomes to two years. AB - OBJECTIVE: To examine the prevalence of supplement use in persons before receiving hematopoietic stem cell transplant (HSCT) and the association of select supplements with outcomes. DESIGN: This observational cohort study included a questionnaire on supplement use before HSCT. Nonrelapse mortality, recurrence/relapse, and mortality or relapse (the inverse of disease-free survival) were followed to two years. Subjects/Setting Persons receiving HSCT at the Fred Hutchinson Cancer Research Center between September 1994 and December 1997 were eligible (N=1,182). Statistical Analyses Performed Descriptive statistics and univariate and Cox regression analyses were conducted. RESULTS: Sixty-six percent of patients used supplements (31% vitamin C, 19% vitamin E, and 20% herbs or others preparations). Vitamin C at > or =500 mg/day was inversely associated with recurrence among persons with breast cancer (RR=0.11; 95% CI, 0.02-0.89; P=.03). However, among persons with acute leukemia, vitamin C at > or =500 mg/day was positively associated with nonrelapse mortality (RR=2.25; 95% CI, 1.33-3.83; P=.01) and mortality or relapse (RR=1.63; 95% CI, 1.09-2.44; P=.01), respectively. Vitamin E at > or =400 IU/day was positively associated with mortality or relapse (RR=1.77; 95% CI, 1.06 -2.96; P=.02). Applications/Conclusions Though this work was observational, the results suggest supplemental vitamin C before therapy may be beneficial in persons with breast cancer but both vitamin C and vitamin E may increase risk in persons with acute leukemia receiving HSCT. Practitioners should document supplement use in subjects receiving therapy for cancer. PMID- 12891147 TI - Folate intake assessment: validation of a new approach. AB - OBJECTIVE: To describe the performance characteristics and validate a folate specific focused recall approach to estimating folate intake. DESIGN: In a cross sectional study, folate-specific recalls were used to estimate subjects' average daily folate intake from seven days of dietary recall over a one-month period. This estimate was compared to an estimate of the average daily folate intake obtained using the Fred Hutchinson Cancer Research Center Food Frequency Questionnaire (FFQ) and to plasma folate and homocysteine concentrations. SUBJECTS: Subjects were women aged 21 to 47 years and were recruited from the university community. Thirty-three subjects were recruited, and 28 subjects completed the study. INTERVENTION: After an initial clinic visit, during which a baseline blood sample was collected for measurement of plasma folate and total homocysteine concentrations, subjects participated in focused dietary recall interviews by telephone and/or Internet (e-mail). At the end of the month, subjects completed the FFQ. Main outcome measures The main outcome measures were to describe how well the focused recalls were able to describe relative dietary folate intake as compared to the FFQ and if the focused recall method estimate correlated with plasma folate and homocysteine concentrations. Statistical analyses Estimated folate intakes were ranked by tertiles using the two dietary assessment approaches and the rankings were compared. The Spearman correlation test was used to compare the focused recall and the FFQ dietary data to plasma folate and homocysteine concentrations. RESULTS: Estimated folate intake inclusive of supplements was 411.4+/-192.8 (mean+/-SD) microg/day based on focused recalls and 458.5+/-221.5 microg/day based on the FFQ. Exclusive of supplements, estimated folate intake from the diet was 327.8+/-30.2 microg/day based on focused recalls and 373.5+/-155.6 microg/day based on the FFQ. The ranking comparison of focused recalls vs the FFQ demonstrated a concordance of 44% without supplements and 67% with supplements for the highest tertile, and 56% without supplements and 67% with supplements for the lowest tertile. Estimated folate intake based on focused recalls was correlated significantly with plasma homocysteine (r=-0.443, P<.02) and was marginally significantly correlated with plasma folate (r=0.354; P=.06) concentrations. APPLICATIONS: A focused dietary recall approach is a useful method for collecting information on folate intake in women. Estimated folate intake based on the focused recall approach is reasonably correlated with plasma folate and total homocysteine concentrations even in this small sample of women. Accurate data on folate intake can form the basis of dietary counseling to increase intake of this important micronutrient in the target population of young women. PMID- 12891148 TI - Comparison of estimated renal net acid excretion from dietary intake and body size with urine pH. AB - OBJECTIVE: The aim of this study was to examine whether estimated net acid excretion (NAE) from two dietary assessment instruments can predict urine pH in healthy volunteers. DESIGN: The Observing Protein and Energy Nutrition (OPEN) study, which was designed to assess dietary measurement error, included men and women aged 40 to 69 years. Participants were asked to complete two food frequency questionnaires (FFQs) and two 24-hour recalls at approximately three-month intervals, and to return two 24-hour urine samples during the same period. NAE was estimated using a previously described equation. The estimates obtained from the different dietary assessment measures were correlated with actual urine pH. SUBJECTS/SETTING: Four hundred eighty-four subjects were enrolled in the OPEN study. All participants were included in this study on urine pH, given that they had all completed at least one dietary assessment and one urine sample. The study took place in Montgomery County, MD, between September 1999 and March 2000. RESULTS: Correlation coefficients between NAE estimates using the FFQ data and the average of two urine pH measures were -0.34 and -0.39 for women and men, respectively. Removing within-person variance in the urine pH levels resulted in a deattenuated overall correlation of -0.50 for the FFQ. Correlations between the 24-hour recall estimates of NAE were -0.41 and -0.32 for women and men, respectively. For the 24-hour recalls, the deattenuated overall correlation was 0.50. APPLICATION/CONCLUSIONS: Our findings suggest that FFQs provide adequate dietary data to estimate renal NAE and could be used as a surrogate for urinary pH levels in large epidemiologic studies. PMID- 12891149 TI - Ethnic differences in the nutrient intake adequacy of premenopausal US women: results from the Third National Health Examination Survey. AB - OBJECTIVE: To examine the adequacy of dietary intake of calcium; folate; and vitamins C, D, E, B-6, and B-12 in premenopausal US women of differing ethnicity. DESIGN: Analyses of single and duplicate 24-hour recalls were conducted to determine dietary intake during the Third National Health and Nutrition Examination Survey. SUBJECTS: Three thousand five hundred eighty-five randomly selected women aged 20 to 50 years from across the United States who were not pregnant or lactating were examined between 1988 and 1994. STATISTICAL ANALYSES: Usual nutrient intake distributions were estimated using the Iowa State University method for adjustment of the distribution. The Estimated Average Requirement cut-point method was used to determine the proportion of women with inadequate intake for each nutrient in each ethnic group. RESULTS: More than 75% of women irrespective of ethnic group had usual intakes of calcium lower than the new Adequate Intake. More than 90% of the women had inadequate intakes of folate and vitamin E from food sources alone. More than half of smokers had inadequate intakes of vitamin C. Intakes of vitamins B-6 and B-12 were low in less than 10% of these women. APPLICATIONS/CONCLUSIONS: This article provides evidence that a high proportion of premenopausal US women are underconsuming a variety of nutrients. Dietary intakes alone are not currently adequate to meet the new recommended intakes. Nutritional supplement use is widespread and effective, but does not eliminate the concerns for at-risk populations. Awareness of the general inadequacies in intakes of vitamin E and folic acid at large, and in many women vitamin C as well, can help direct individual dietary recommendations and place the emphasis in group counseling on nutrients that are of widespread concern. In addition, foods rich in vitamins B-6 and of general nutritional benefit should be emphasized among African American women in the United States as a substantial proportion of this group is still showing inadequate intakes from foods. PMID- 12891150 TI - Dietary intake and food sources of whole grains among US children and adolescents: data from the 1994-1996 Continuing Survey of Food Intakes by Individuals. AB - OBJECTIVE: This study characterizes whole grain consumption among a nationally representative sample of US children and adolescents. DESIGN: Data used in this study were collected as part of the 1994-1996 US Department of Agriculture Continuing Survey of Food Intakes by Individuals (CSFII). SUBJECTS/SETTING: CSFII was designed to obtain a nationally representative sample of noninstitutionalized persons of all ages residing in the United States. Analyses reported in this article are limited to participants aged 2 to 18 years with two days of dietary recall data (n=4,802). Foods reported in the survey were quantified in servings as defined by the Food Guide Pyramid using the US Department of Agriculture Pyramid Servings Database, which contains reference data for each food reported in CSFII in servings per 100 g for 30 Pyramid food groups, including whole grain and total grain. STATISTICAL ANALYSES: Means, frequencies, and logistic regression analyses were conducted as appropriate. RESULTS: Average whole grain intake ranged from 0.8 servings per day for preschool-aged children to 1.0 servings per day for adolescents. Ready-to-eat cereals, corn and other chips, and yeast breads were found to be the major food sources of whole grains accounting for 30.9%, 21.7%, and 18.1% of whole grain intake respectively among those aged two to 18 years. APPLICATIONS/CONCLUSIONS: Given the apparent low level of whole grain intake among most children and adolescents in the United States, interventions are needed to increase intake of whole-grain foods. PMID- 12891152 TI - Introducing a problem-based unit into a lifespan nutrition class using a randomized design produces equivocal outcomes. AB - Problem-based learning (PBL) is a tutorial, student-centered, problem-driven educational strategy adopted by medical and allied health educators to positively influence self-directed learning, critical thinking, and learning behavior. PBL was examined in dietetics education through random assignment of 32 undergraduate dietetics students for two weeks to either a problem-based or a lecture-based case format for the infant and elderly units of a 16-week lifespan nutrition course. Random assignment followed stratification for gender and earlier course examination performance. Expert validation of PBL curricular components and noted differences in discussion structure and information resources verified curriculum distinctiveness. Main outcome measures were pre- and post-Cognitive Behavior Survey scores for memorization, reflection, and positive learning experience scales, unit and course evaluations and unit examination scores. Students in problem-based modules demonstrated greater gains in reflective thinking with stable memorization, suggesting improved critical thinking skills. Tenets that problem-based learning promotes knowledge retention and provides a more positive learning experience were not upheld. Knowledge acquisition was not hindered by a problem-based approach. We conclude that gains in reflective thinking and evidence of increased self-directed learning argue for inclusion of PBL in dietetics curricula and that a problem-based education will help dietitians successfully respond to professional development needs. PMID- 12891153 TI - Continuous improvement in dietetics education with a regional advisory board: a model that works. AB - Continuous improvement has been a focus of business and health care for years. The Commission on Accreditation for Dietetics Education has also identified the need for continuous improvement in dietetics education programs and requires them to seek outside counsel to help accomplish it. Most dietetics education programs develop advisory boards to provide this outside counsel, but finding individuals to commit to an ongoing advisory board can be challenging. Dietetics internship directors from dietetics practicum programs throughout Arizona created the Arizona Dietetic Practicum Advisory Committee (AzDPAC), which uses suggestions from preceptors, interns, and committee members to generate ideas for improvement. Since the conception of AzDPAC both tangible and intangible outcomes have occurred. Tangible outcomes include better coordination among programs, development of a combined annual affiliate meeting, standardization of forms, creation of a combined "Clinical Bootcamp," development of a Web page describing all programs, and better networking among interns from different programs. In addition, AzDPAC improved cooperation and sharing of expertise and created an available group of peers for new directors. An advisory committee of regional program directors is a model that works in providing dietetics education programs with ongoing outside counsel and ideas for continuous improvement. PMID- 12891155 TI - Prevalence of outsourcing and perception of clinical nutrition managers on performance of health care dietetics services. AB - A nationwide survey of clinical dietitians and clinical nutrition managers was conducted to assess the prevalence of outsourcing in health care dietetics services and to evaluate perceived performance of dietetics services. A questionnaire was developed, validated by an expert panel, and pilot tested prior to data collection. Members of the Clinical Nutrition Management Dietetic Practice Group (N=1,668) were selected as the study sample. Of 431 respondents, 152 (35.3%) indicated that management of both patient and cafeteria foodservices was outsourced. When mean scores of perceived performance ratings were compared using t test, respondents from self-operated facilities rated several items related to patient and cafeteria food quality and material and human resource utilization higher than respondents at contract-managed facilities. No significant differences were found in performance related to decision-making process, buying power, or training programs. Results suggest that careful weighing of advantages and disadvantages of outsourcing is needed before making decisions regarding outsourcing dietetics services. PMID- 12891154 TI - Nutritional cofactor treatment in mitochondrial disorders. AB - Mitochondrial disorders are degenerative diseases characterized by a decrease in the ability of mitochondria to supply cellular energy requirements. Substantial progress has been made in defining the specific biochemical defects and underlying molecular mechanisms, but limited information is available about the development and evaluation of effective treatment approaches. The goal of nutritional cofactor therapy is to increase mitochondrial adenosine 5' triphosphate production and slow or arrest the progression of clinical symptoms. Accumulation of toxic metabolites and reduction of electron transfer activity have prompted the use of antioxidants, electron transfer mediators (which bypass the defective site), and enzyme cofactors. Metabolic therapies that have been reported to produce a positive effect include Coenzyme Q(10) (ubiquinone); other antioxidants such as ascorbic acid, vitamin E, and lipoic acid; riboflavin; thiamin; niacin; vitamin K (phylloquinone and menadione); creatine; and carnitine. A literature review of the use of these supplements in mitochondrial disorders is presented. PMID- 12891156 TI - Prevention of childhood obesity: sociocultural and familial factors. AB - This study examined sociocultural and familial factors related to the prevention of childhood obesity. Primary caregivers of 6- to 10-year-old children representing several ethnic populations in Saipan participated in 4 focus groups (N=32). Trained moderators used semi-structured interviews and qualitative methods were used in data analysis. A central theme with several related factors emerged. The theme was a conflict expressed by the primary caregiver between sociocultural values, family expectations, traditional dietary beliefs and attitudes, and knowledge about food and disease. These findings have important implications for designing culturally sensitive interventions for prevention of childhood obesity. PMID- 12891157 TI - Position of the American Dietetic Association: Addressing world hunger, malnutrition, and food insecurity. AB - It is the position of the American Dietetic Association (ADA) that access to adequate amounts of safe, nutritious, and culturally appropriate food at all times is a fundamental human right. Hunger continues to be a worldwide problem of staggering proportions. The Association supports programs and encourages practices that combat hunger and malnutrition, produce food security, promote self-sufficiency, and are environmentally and economically sustainable. The Association is aware that hunger exists in a world of plenty and that poverty, gender inequity, ethnocentrism, racism, and the lack of political will are key constraints to solving the problems of global hunger and malnutrition. Recognizing that simplistic approaches are inadequate, the ADA identifies sustainable development as the long-term strategy to ending world hunger and achieving food security. Sustainable development requires political, economic, and social changes that include empowering the disenfranchised, widening access to assets and other resources, narrowing the gap between rich and poor, and adjusting consumption patterns so as to foster good stewardship of nature. Additionally, because the health status of future generations is related to the well-being of their mothers, achieving food security will also require increased access for women to education, adequate health care and sanitation, and economic opportunities. This position paper reviews the complex issues of global food insecurity and discusses long-term solutions for achieving world food security. Achieving the end of world hunger has been and is now within our grasp. There is sufficient food to feed everyone, and solutions can be realized now that will benefit all of humanity. As noted in the paper, most people who examine the costs of ending versus not ending world hunger are bewildered by the question of why humanity did not solve the problem a long time ago. The Association supports programs and encourages practices that combat hunger and malnutrition, produce food security, promote self-sufficiency, respect local cultures, and are environmentally and economically sustainable. The ADA recognizes that decisions and actions that dietetics professionals make as practitioners and consumers can help reduce the extent of poverty and hunger both here and abroad. This paper provides information, resources, and strategies to assist dietetics professionals in improving the public's understanding of key issues, becoming advocates of the poor, and influencing the political will to end world hunger. PMID- 12891158 TI - Sugars and health: is there an issue? PMID- 12891159 TI - Nutrition Care Process and Model: ADA adopts road map to quality care and outcomes management. PMID- 12891160 TI - Sex-related differences in heart failure. AB - Heart failure (HF) is a complex syndrome that is generally defined as cardiac output not adequate to meet the circulatory demands of the body. HF is at the end of the continuum of cardiovascular disease and preceded by an initiating event such as myocardial infarction, untreated hypertension, idiopathic cause, congenital heart disease, or pulmonary hypertension. In recent years, research has revealed differences in various aspects of HF between men and women including risk factors, pathophysiology, clinical manifestations, and response to treatment. Therefore, the purpose of this review is to review these sex-related differences between men and women who live with HF. PMID- 12891162 TI - Pilot study of a Web-based compliance monitoring device for patients with congestive heart failure. AB - BACKGROUND: Web-based home care monitoring systems can assess medication compliance, health status, quality of life, and physiologic parameters. They may help overcome some of the limitations associated with current congestive heart failure management models. OBJECTIVES: This pilot study compared the effects of a self-care and medication compliance device, linked to a Web-based monitoring system, to the effects of usual care alone on compliance with recommended self care behaviors; medication taking; quality of life; distance walked during a 6 minute walk test; and New York Heart Association Functional Class. We also assessed patient experiences living with the compliance device. METHODS: We enrolled 18 patients with Functional Class II-III congestive heart failure in an urban VA Medical Center. The patients were randomized into 2 groups. Group A received usual care plus the compliance device. Group B (controls) received usual care only. Data were collected using the compliance device, the Heart Failure Self-Care Behavior Scale, pill counts, 6-minute walk test, and the Minnesota Living with Heart Failure Questionnaire at baseline and at 3 months follow-up. RESULTS: At baseline and at 3 months, there were no differences between the compliance device group and the usual care group in self-care behaviors, pill counts, 6-minute walk-test distance, or Functional Class. However, quality of life improved significantly from baseline to 3-month follow-up (ANOVA, P =.006). This difference was due to an improvement in quality of life for the monitor group (P =.002) but not the usual care only group (P =.113). Patients in the compliance device group had a 94% medication compliance rate, 81% compliance with daily blood pressure monitoring, and 85% compliance with daily weight monitoring as compared to 51% for blood pressure monitoring and 79% for weight monitoring in the usual care group (P = NS). CONCLUSION: These are promising pilot results that, if replicated in a larger sample, may significantly improve care and outcomes for patients with heart failure. PMID- 12891163 TI - Psychological therapies for recipients of implantable cardioverter defibrillators. AB - Implantable Cardioverter Defibrillators (ICDs) are an increasingly common therapy for the treatment of ventricular arrhythmias. Whereas most ICD recipients adjust well to living with the implant, a substantial minority suffer anxiety, depression, and anger that appears to be related to the ICD. A large number of studies have reported on the incidence of psychopathology and common psychosocial problems among patients with ICDs, however very few psychological interventions with this cohort have been reported. In an extensive literature search we identified only a handful of studies that described outcomes of psychological interventions, and most of these were in pilot form. Only one randomized controlled study with a homogenous sample of ICD patients has been reported to date. Given the unique situation of patients with ICDs and the particular vulnerability of those who experience frequent shocks, the evaluation of interventions that may improve psychological adjustment within this group would appear a worthwhile endeavor. PMID- 12891164 TI - A descriptive study of early nonspecific chest pain after PTCA: important area for the acute health care personnel. AB - BACKGROUND: Many patients report chest pain of varying intensity at various locations during the first hours after percutaneous transluminal coronary angioplasty (PTCA). OBJECTIVES: The aim of the study was to increase knowledge regarding differentiating between harmless chest pain versus ischemic chest pain, focusing on patients description of their pain. METHODS: A total of 192 patients after elective PTCA were interviewed twice. In addition patients experiencing chest pain within 6 hours after the procedure completed the McGill Pain Questionnaire (MPQ). RESULTS: Nonspecific chest pain occurred in 34 patients (18%) and ischemic chest pain in 6 (3%), whereas 152 (79%) did not report early chest pain after PTCA. The nonspecific pain group reported statistically significant less pain intensity (VAS P =.001), used fewer (P =.006) and qualitatively weaker (P =.008) words compared to the ischemic pain group. No predisposing factors that could predict chest pain were identified. CONCLUSIONS: Discriminators appear to be the pain intensity and the word descriptors. MPQ combined with a VAS could be valuable clinical tools with regard to patients' description of pain. PMID- 12891165 TI - Meaning and life purpose: the perspectives of post-transplant women. AB - PURPOSE: A descriptive, exploratory study was conducted to examine perceptions and meanings assigned to the experience of end-stage heart failure and transplant surgery in female recipients. METHODS: Data was collected from 33 female heart transplant recipients from a heart transplant clinic using quantitative and qualitative methods. Women completed the Life Attitudes Profile and Multiple Affect Adjective Checklist and participated in a semi-structured interview. RESULTS: Women on average had a meaning score of 30.64 +/- 8.32 (range, 10-43) and a life purpose score of 40.12 +/- 13.78 (range, 17-63). The participants reported high levels of anxiety, depression, and hostility. Women expressed feelings of fear and lack of control related to their heart disease and transplant; however, they also described optimism, faith, acceptance, altruism, self-transcendence, self-fulfillment, and changing life goals as strong motivators for recovery. CONCLUSION: Women experience emotional distress as much as and beyond 5 years after heart transplantation. Their sense of meaning and life purpose is relatively high and is positively informed by several motivating factors that serve as coping resources throughout the recovery process. PMID- 12891167 TI - Case report: Acute myocardial infarction in a 32-year-old white male found to have antiphospholipid antibody syndrome and MTHFR mutation homozygosity. AB - We report the diagnosis and management of a 32-year-old Hungarian male, whose only known risk factor for coronary artery disease was smoking, who presented with an acute thrombotic anterolateral wall myocardial infarction requiring percutaneous transluminal coronary angioplasty (PTCA) stenting of his proximal left anterior descending coronary artery. He arrived to the emergency room with an abnormally prolonged partial thromboplastin time (PTT) that subsequently did not correct by mixing with normal plasma. This was suggestive of an underlying coagulopathy. An extensive coagulopathy work up found him to have the antiphospholipid antibody syndrome with antibodies positive for anticardiolipin, lupus anticoagulant and false-positive VDRL. Genetic typing found him to be homozygous for a mutation in the methylenetetrahydrofolate reductase (MTHFR A1298C) gene, which, in the presence of additional thrombophilic factors, may have increased his risk of myocardial infarction. He was discharged on high dose coumadin. PMID- 12891166 TI - Aerobic exercise and the post myocardial infarction patient: a review of the literature. AB - Meta analyses of randomized controlled tests of cardiac rehabilitation after myocardial infarction demonstrate that regular exercise reduces the risk of overall mortality and cardiovascular mortality. In patients with established coronary artery disease, exercise is associated with improved activity tolerance, modification of risk factors, and improvement in quality of life. Randomized controlled tests demonstrate that whereas older patients after coronary events are substantially less fit than younger patients, they obtain a similar relative improvement of aerobic capacity with a graded conditioning program. However, older adults are enrolled in such programs at a lower rate than other age groups. Despite similar clinical profiles to men, women are less likely to participate in exercise rehabilitation. In this article we discuss the principles of program development, guidelines for monitoring of patients, and facilitation of exercise programs in the Australian context. PMID- 12891168 TI - Native aortic valve enterococcal endocarditis with myocardial paravalvular abscess. PMID- 12891169 TI - Description of anxiety levels by individual differences and clinical factors in patients receiving mechanical ventilatory support. AB - BACKGROUND: Though anxiety is a common experience for patients receiving mechanical ventilatory support, little is known about how it may vary among patients on the basis of individual or clinical factors. There is an absence of data objectively describing anxiety levels in ventilated patients on the basis of salient factors that could be useful in designing and tailoring interventions. PURPOSE: The purpose of this study was to describe anxiety levels in a sample of mechanically ventilated patients by individual differences (eg, gender or ethnicity) and clinical factors (eg, medical indication for and length of mechanical ventilation). SAMPLE: Two hundred alert, mechanically ventilated adult patients were recruited from 9 intensive care units in the urban Midwest. METHODS: This study was a secondary analysis of existing data that used a descriptive design. Anxiety was assessed via the 20-item Spielberger State Anxiety Inventory. RESULTS: Whereas state anxiety varied widely, participants receiving mechanical ventilatory support reported moderate anxiety (mean = 49.2) with comparable levels by gender and ethnicity. Patients receiving ventilatory support for greater than 22 days tended to report slightly higher state anxiety (mean = 54.2) compared with those chronically ventilator dependent (mean = 45.8). Those participants with primarily respiratory diagnoses reported the highest levels (50.5) among the diagnostic groups. Findings from this study document the individual, variable nature of state anxiety. Additional research is needed to further elucidate whether these and other important clinical factors, such as illness severity or dyspnea, affect state anxiety ratings in ventilated patients to guide the researcher and clinician in appropriately testing and tailoring interventions. PMID- 12891170 TI - Effects of age and race on skin condition and bacterial counts on hands of neonatal ICU nurses. AB - OBJECTIVE: The purpose of this study was to assess the relationship between demographic factors such as age and race and skin condition and bacterial counts on hands of nurses. METHODS: Nurses (n = 111) working in 1 of 2 neonatal ICUs in New York City were surveyed regarding reported hand care practices and demographics, the condition of their hands was assessed by a trained observer and by themselves using validated instruments, and a hand culture was obtained. RESULTS: There were no significant differences in any skin care practices by race, but nurses who wore powder-free gloves were significantly younger than those who did not (P =.004). There were no significant differences in bacterial counts on hands of black or white nurses (mean log colony-forming units 3.49 and 3.61 respectively, P =.63) and no significant correlation between age and microbial counts (r = 0.04, P =.72). In a logistic regression analysis, race, but not age, was a significant predictor of skin health. By both observer (P =.02) and self-assessment (P =.004) black nurses had healthier skin. CONCLUSION: Physiochemical differences in skin associated with demographic factors such as age and race may be exacerbated among those in disciplines such as nursing, for whom the skin of the hands is continually stressed by occupational practices such as frequent hand hygiene. Such demographic factors need be considered when assessing skin condition and when advocating for appropriate strategies to maximize skin health. The recommendations of the new CDC Hand Hygiene Guideline for Healthcare Settings(23) regarding maintaining skin health and providing moisturizers and products that are milder to the skin are timely and should be followed. PMID- 12891178 TI - Vegetations in endocarditis: big is bad, but is there more to it? PMID- 12891179 TI - Low molecular weight heparins for acute coronary syndrome: tackling the issues head-on. PMID- 12891180 TI - Heart failure and the elderly: disease diversity, data, and delivery of care. PMID- 12891181 TI - Are heart rate and blood pressure responses to mental stress predictive of clinical cardiovascular events? The answer varies by coronary artery disease syndrome. PMID- 12891182 TI - Inflammation in atherosclerosis: causal or casual? The need for randomized trials. PMID- 12891183 TI - Does race matter in heart failure? PMID- 12891184 TI - Is geography destiny? Illuminating the survival advantage of elderly patients in New England after acute myocardial infarction. PMID- 12891185 TI - Subclinical diabetes mellitus: is it really "sub-clinical"? PMID- 12891186 TI - Percutaneous coronary interventions in patients with renal insufficiency: a high risk, under-studied cohort. PMID- 12891187 TI - Hippocrates and prognosis in heart failure: the challenge of static and dynamic measurements. PMID- 12891188 TI - Endothelial dysfunction: clinical strategies for treating oxidant stress. AB - BACKGROUND: A growing body of evidence has demonstrated that oxidants play a critical role in the pathogenesis of endothelial dysfunction. Pathologic processes fundamental to development and progression of endothelial dysfunction such as the oxidation of LDL, the loss of bioavailable nitric oxide, and the vascular inflammatory response are all modulated by oxidant stress. Therapeutic strategies to reverse endothelial dysfunction have begun to focus on agents with the ability to ameliorate oxidant stress. METHODS: Preclinical and clinical studies evaluating the actions of antioxidants as well as traditional cardiovascular therapies in ameliorating oxidative stress and endothelial dysfunction were reviewed through the use of a MEDLINE search of English language articles published between the years of 1992 and 2002. RESULTS: Antioxidants appear to be an attractive candidate therapy, yet despite compelling preclinical evidence supporting their benefits, efforts to validate the use of vitamins C and E in a clinical setting have been conflicting. In contrast, conventional cardiovascular therapies such as ACE inhibitors, statins, insulin-sensitizing agents, and estrogens have been shown to alleviate endothelial dysfunction, often independent of their effects on systemic disease processes. CONCLUSIONS: These agents restore endothelial function through their salutary effects on pathologic vascular oxidative processes. PMID- 12891189 TI - Role of lipid and lipoprotein profiles in risk assessment and therapy. AB - Although low-density lipoprotein cholesterol (LDL-C) remains the primary target for coronary heart disease (CHD) prevention in the latest guidelines of the National Cholesterol Education Program, many individuals who have CHD do not have substantially elevated LDL-C but have derangement of other lipid fractions, most commonly low levels of high-density lipoprotein cholesterol (HDL-C). In the guidelines, HDL-C is important in risk stratification in primary prevention, influencing the need for and intensity of treatment of LDL-C, and both HDL-C and triglyceride are defined as risk factors for the metabolic syndrome, a secondary target of therapy. Triglyceride level also determines in which individuals non HDL-C should be a secondary target of therapy. Risk assessment that takes into account the entire lipid profile will identify more high-risk individuals than evaluating LDL-C alone. Some epidemiologic data suggest that instead of measuring the cholesterol in LDL or HDL, measuring their respective apolipoproteins, apolipoprotein (apo) B-100 and apo A-I, may improve CHD risk assessment, and in some observational and interventional studies, ratios of lipids and/or apolipoproteins have been better predictors of CHD risk than levels of any one lipid fraction. Trials of lipid-modifying therapy also suggest that apolipoproteins and ratios may provide improved targets for therapy beyond LDL-C, but optimal values have not been established. Because lipid-modifying therapy affects multiple components of the lipid profile, the effect on all lipid parameters should be considered when selecting the most appropriate agent. Therapies with beneficial effects across the lipid profile would be expected to improve CHD risk reduction. PMID- 12891190 TI - Danish multicenter randomized study on fibrinolytic therapy versus acute coronary angioplasty in acute myocardial infarction: rationale and design of the DANish trial in Acute Myocardial Infarction-2 (DANAMI-2). AB - BACKGROUND: Randomized trials have indicated that primary coronary angioplasty performed in patients admitted directly to highly-experienced angioplasty centers offers certain advantages over intravenous fibrinolytic therapy. However, the large majority of patients with acute myocardial infarction are submitted to hospitals without a catheterization laboratory. This means that additional transportation will be necessary for many patients if a strategy of acute coronary angioplasty is to be introduced as routine treatment. The delay of treatment caused by transportation might negate (part of) the benefits of primary angioplasty compared to fibrinolytic therapy given immediately at the local hospital. STUDY DESIGN: The DANish trial in Acute Myocardial Infarction-2 (DANAMI 2) is the first large-scale study to clarify, in a whole community, which of the 2 treatment strategies is best. A total of 1900 patients with ST-elevation myocardial infarction are to be randomized: 800 patients will be admitted to invasive hospitals and 1100 patients will be admitted to referral hospitals. Half of the 1100 patients admitted to referral hospitals will immediately be transferred to an invasive center to be treated with primary angioplasty. IMPLICATIONS: If acute transfer from a local hospital to an angioplasty center is the superior strategy, primary angioplasty should be offered to all patients as routine treatment on a community basis. PMID- 12891191 TI - Regional variation in the treatment and outcomes of myocardial infarction: investigating New England's advantage. AB - BACKGROUND: Previous studies have reported that myocardial infarction (MI) treatment in New England differs from that of other regions of the United States. We sought to determine whether regional differences in MI treatment were independent of regional differences in patient, hospital, or physician characteristics, and whether the New England region's practice pattern was associated with better outcomes than those of patients in other regions. METHODS: We evaluated 167,180 patients aged > or =65 years who were hospitalized with MI between 1994 to 1996 to assess regional variations in quality of care. Patients were evaluated for the use of reperfusion therapy, aspirin, and beta-blockers on admission and 30-day mortality rate. Hierarchical logistic regression models were used to determine whether practice patterns specific to New England were independent of regional variations in patient, physician, hospital, or other geographic characteristics. RESULTS: New England had the highest use of beta blockers (72% vs 52% other regions, P <.001), and aspirin (80% vs 76% other regions, P <.001), a lower use of reperfusion therapy (61% vs 67% other regions, P <.001), and the lowest risk-standardized 30-day mortality rate (15% vs 19% other regions, P <.001). These differences persisted after adjusting for patient, physician, and hospital characteristics. CONCLUSIONS: Patients with MI in New England have higher rates of medical therapy use and lower 30-day mortality rates than patients in other US regions. This pattern is independent of patient or provider characteristics, suggesting other factors likely contribute to better short-term outcomes in New England. PMID- 12891193 TI - Mortality trends for 23,505 Medicare patients hospitalized with heart failure in Northeast Ohio, 1991 to 1997. AB - BACKGROUND: Clinical trials have identified major therapeutic advances for heart failure (HF), but the degree to which survival has improved among the general population of patients with HF is not known. This study analyzed mortality trends from 1991 to 1997 for 23,505 Medicare patients hospitalized with a first admission for HF at 29 Northeast Ohio hospitals. METHODS: We linked databases from the Cleveland Health Quality Choice (CHQC) program and Medicare to allow identification of first admissions for HF and death date. We adjusted for changes in admission illness severity using chart data from CHQC (eg, vital signs, do-not resuscitate status, comorbid conditions, and laboratory results). Logistic regression was used to analyze trends in risk-adjusted mortality. RESULTS: At baseline (1991), crude inhospital, 30-day and 1-year mortality rates were 6.4%, 8.6% and 36.5%, respectively. Between 1991 and 1997, mean length of stay declined steeply from 9.2 days to 6.6 days (P <.001 for trend). Risk-adjusted inhospital mortality also declined markedly (absolute-decline -3.7%, 95% CI -4.3 to -3.0), a 52.8% relative decrease. However, the decline in 30-day mortality was only -1.4% (95% CI -2.5 to -0.1, P <.05), a 15.3% relative decrease. The 1-year mortality declined -5.3% (95% CI -3.2 to -7.4, P <.001), a 14.6% relative decrease. CONCLUSIONS: Long-term mortality for patients hospitalized with HF improved from 1991 to 1997, although mortality remains very high. The 30-day mortality declined far less than inhospital mortality, indicating that mortality shortly after discharge increased. This raises concerns that the marked reduction in length of stay is causing adverse consequences. PMID- 12891192 TI - Most hospitalized older persons do not meet the enrollment criteria for clinical trials in heart failure. AB - BACKGROUND: Although it is widely accepted that clinical trials in heart failure may not apply to older populations, the magnitude of the discrepancy between trial populations and patients seen in community-based practice are not known. Our objective was to determine the proportion of older persons meeting enrollment criteria of randomized controlled trials of agents that prolong life in heart failure. METHODS: We conducted a cross-sectional study of Medicare beneficiaries >64 years old with the principal diagnosis of heart failure who were discharged from acute care hospitals in the United States between April 1998 and March 1999. Enrollment criteria of the Studies of Left Ventricular Dysfunction (SOLVD), Metroprolol CR/LX Randomized Intervention Trial in Congestive Heart Failure (MERIT-HF), and Randomized Aldactone Evaluation Study (RALES) trials were applied to the population, and the proportions meeting the criteria were determined by subgroups of age and sex. RESULTS: Of the 20,388 patients studied, 18%, 13%, and 25% met the enrollment criteria of the SOLVD, MERIT-HF, and RALES trials, respectively. Although trial eligibility was less than a third for any sex or age group, significantly fewer women than men met trial criteria (13% vs 23% for SOLVD, 11% vs 17% for MERIT-HF, and 21% vs 32% for RALES, P <.0001 for all). The oldest patients were also less likely to fulfill enrollment criteria. The proportion of all patients not included because of preserved left ventricular systolic function was twice as large as the proportion meeting the inclusion criteria for any trial. CONCLUSIONS: A minority of hospitalized older persons with heart failure fit the profile of populations of clinical trials. There is an urgent need for research in heart failure for typical heart failure patients, including the very old, women, and patients with preserved left ventricular systolic function. PMID- 12891194 TI - Survival rates are similar between African American and white patients with heart failure. AB - BACKGROUND: The clinical characteristics of heart failure differ significantly between African American patients and white patients, apparently as a result of differences in the pathobiology of the condition in the races. We investigated the hypothesis that race also influences the survival of patients with heart failure. METHODS: Data from the University of North Carolina Heart Failure Database were analyzed for 853 patients (44% African American, 32% women) who had symptomatic heart failure (New York Heart Association class 2.8 +/- 0.02 [mean +/ SEM]) with a reduced left ventricular ejection fraction of 26% +/- 0.5% and a body mass index of 27 +/- 0.2. Data on vital status were available in 96.4% of these patients, with a mean length of follow-up of 3.8 +/- 0.1 years. RESULTS: An unadjusted univariate proportional-hazards analysis suggested similar survival rates between African American patients and white patients in the study population (relative risk, 0.90; 95% CI, 0.73-1.10; P =.293). Adjusted analysis, taking into account the characteristics shown to be of prognostic importance, demonstrated no difference in survival rate between African American patients and white patients (relative risk,1.12; 95% CI, 0.89-1.42; P =.336). The adjusted relative risk of all-cause mortality in the respective races among patients with heart failure caused by ischemic heart disease was 1.21 (95% CI, 0.80-1.84; P =.367). CONCLUSION: African American and white patients with symptomatic heart failure had similar survival rates in our database. PMID- 12891195 TI - Heart rate and blood pressure responses to mental stress and clinical cardiovascular events in men and women after coronary artery bypass grafting: the Post Coronary Artery Bypass Graft (Post-CABG) biobehavioral study. AB - BACKGROUND: The Post-Coronary Artery Bypass Graft (CABG) biobehavioral study measured the heart rate (HR) and blood pressure (BP) responses to mental stress testing (MST) after surgery and prospectively observed the clinical events in patients who had undergone recent CABG surgery. To the extent that CABG surgery restores myocardial blood flow and prevents myocardial ischemia, patients who have recently recovered from CABG surgery may have nearly normal HR and BP responses to MST. METHODS: A study population of 521 patients (351 men,170 women) from the cohort of 759 patients in the Post-CABG biobehavioral Study was monitored during a mirror tracing test (MTT) and a speech test task (STT) at the 6-month post-CABG surgery follow-up. Medical status for as long as 3 years after CABG surgery was ascertained from questionnaires and medical records. RESULTS: The HR and BP responses during MTT and STT were similar to those reported in other studies of healthy individuals and patients without myocardial ischemia during MST. In contrast to results from other studies of patients with coronary artery disease, the quartile of Post-CABG biobehavioral study patients with the greatest responses in HR, systolic BP (SBP) and diastolic BP (DBP) during MST had half the rate of clinical events (10% in 3 years) as the quartile (20%) with the lowest changes in HR and BP (HR, P =.01; SBP, P =.07; DBP, P =.01). CONCLUSIONS: Vigorous HR and BP responses to MST may be markers for a lower risk of incidence of clinical cardiovascular events among patients who have undergone recent CABG surgery. PMID- 12891196 TI - Improvement of peripheral endothelial dysfunction by acute vitamin C application: different effects in patients with coronary artery disease, ischemic, and dilated cardiomyopathy. AB - BACKGROUND: Endothelial dysfunction has been described in patients with coronary artery disease (CAD) or chronic heart failure (CHF). Vitamin C administration leads to an improvement of endothelial function by reducing elevated levels of reactive oxygen species. It remains unclear, however, whether the degree of endothelial dysfunction caused by oxidative stress differs between CAD and CHF because of ischemic (ICM) or dilated cardiomyopathy (DCM). METHODS: In patients with CAD (n = 9; left ventricular ejection fraction [LVEF], 64% +/- 3%), ICM (n = 9; LVEF, 25% +/- 4%), DCM (n = 9; LVEF, 25% +/- 3%), and healthy subjects (HS; n = 5; LVEF, 66% +/- 5%) a change in internal radial artery diameter in response to acetylcholine (Ach; 15 and 30 microg/min) was measured with high-resolution ultrasound scanning during a co-infusion of normal saline or vitamin C (25 mg/min). RESULTS: Ach-mediated vasodilation was blunted in patients with CHF (DCM, 90 +/- 20 microm; ICM, 86 +/- 20 microm) and patients with CAD (336 +/- 20 microm) as compared with HS (496 +/- 43 microm; P <.05 vs patients with DCM, ICM, CAD). Vitamin C co-infusion increased Ach-mediated vasodilation by 180 +/- 35 microm (to 270 +/- 30 microm) in DCM (P <.05 vs CAD, HS) and by 294 +/- 40 microm (to 380 +/- 20 microm) in ICM (P <.05 vs DCM, CAD, HS). In patients with CAD, vitamin C increased Ach-mediated vasodilation by 146 +/- 35 microm to normal values, whereas vascular diameter remained unchanged in HS (14 +/- 20 microm; P = not significant). CONCLUSIONS: Acute vitamin C administration restored peripheral endothelial function in patients with CAD to normal values, whereas endothelial function remained attenuated in CHF, in particular in patients with DCM. These results suggest that in patients with CHF, factors other than oxidative stress (eg, cytokines) contribute to the pathologic endothelial function. PMID- 12891197 TI - Predictors of mortality in younger and older patients with heart failure and preserved or reduced left ventricular ejection fraction. AB - BACKGROUND: Although half of elderly patients with heart failure have preserved left ventricular ejection fraction (LVEF), little is known about predictors of mortality in this group. METHODS: We reviewed the charts of 400 patients hospitalized at an academic medical center in 1999 with a principal discharge diagnosis of heart failure. Patients were divided into 4 groups on the basis of age > or =75 or <75 years and the presence of preserved or reduced LVEF. Vital status was ascertained as of October 2001. RESULTS: A total of 373 patients (mean age 69.1 years, 56.0% female, 47.5% nonwhite) underwent echocardiography to assess LVEF. Of these, 216 patients were <75 years of age (81 with preserved LVEF [group 1, 21.7%] and 135 with reduced LVEF [group 2, 36.2%]), and 157 were > or =75 years of age (81 with preserved LVEF [group 3, 21.7%] and 76 with reduced LVEF [group 4, 19.6%]). After a mean follow-up of 25 months, independent predictors of mortality among the 4 groups differed substantially: group 1, male sex, prescription of a calcium-channel blocker, and diuretic dose at discharge; group 2, blood urea nitrogen (BUN), lower hemoglobin level, and not being prescribed a beta-blocker at discharge; group 3, BUN; and group 4, older age, history of myocardial infarction, severity of reduced LVEF, and diuretic dose. CONCLUSION: In patients with heart failure, predictors of mortality vary by age and by the presence of preserved or reduced LVEF. Traditional predictors of mortality in patients with reduced LVEF may not apply to elderly patients with preserved LVEF. PMID- 12891198 TI - Neurohormones and oxidative stress in nonischemic cardiomyopathy: relationship to survival and the effect of treatment with amlodipine. AB - OBJECTIVES: The purpose of this study was to assess the effects of amlodipine on neurohormones and oxidative stress in nonischemic cardiomyopathy, and determine the relationship between baseline and posttreatment levels of these markers with survival. BACKGROUND: Neurohormones and oxidative stress are important in the pathophysiology of heart failure. Calcium-channel blockers are associated with poor outcomes in patients with heart failure, in part due to neurohormonal activation. In contrast, amlodipine, a second-generation dihydropyridine, has a more favorable clinical profile. METHODS: In the Prospective Randomized Amlodipine Survival Evaluation 2 (PRAISE-2) trial, a subset of 181 patients with nonischemic cardiomyopathy were randomized to amlodipine (10 mg/day) or placebo. Blood samples were evaluated at baseline, 2 weeks and 26 weeks for norepinephrine, epinephrine, angiotensin II, dopamine, N-terminal pro-atrial natriuretic peptide (Nt-pro-ANP), brain natriuretic peptide (BNP), adrenolutin and malondialdehyde. RESULTS: There was no difference in levels of neurohormones or oxidative stress markers between the amlodipine and placebo groups at the different times. Both Nt-pro-ANP and BNP decreased at 2 weeks and at 26 weeks. Baseline Nt-pro-ANP correlated with survival in multivariate analysis (P =.001). A strong relationship was found between a reduction in BNP at 26 weeks and survival, with a hazard ratio of 0.153 (95% CI 0.051-0.461, P =.017). No relationship was found between markers of oxidative stress and survival. CONCLUSIONS: We conclude that amlodipine does not affect circulating neurohormones and oxidative stress markers in patients with nonischemic cardiomyopathy treated with angiotensin-converting enzyme inhibitors, digoxin and diuretics. In addition, low circulating Nt-pro-ANP and a reduction in BNP over time confers a good prognosis. PMID- 12891199 TI - Serial versus isolated assessment of clinical and instrumental parameters in heart failure: prognostic and therapeutic implications. AB - BACKGROUND: In heart failure (HF), it is not known whether analysis of serial changes in prognostic parameters provides incremental information with respect to comprehensive isolated clinical and instrumental assessments. METHODS: We analyzed time-related changes in a period > or =6 months in a broad panel of clinical and instrumental (electrocardiographic, echocardiographic, hemodynamic, and cardiopulmonary) parameters in 105 patients with HF (age, 53 +/- 10 years; 88% men; 55% New York Heart Association classification III-IV; EF, 24% +/- 6%). RESULTS: Among the time-related parameters, QRS widening (adjusted RR per 10 ms, 1.21; 95% CI, 1.10-1.48; P =.003) and peak oxygen uptake (pVO2) decrease (adjusted RR per mL/Kg/min, 1.11; 95% CI, 1.01-1.22; P =.034) provided independent, incremental information for predicting cardiac death/need for heart transplantation (CD/HT) with respect to the entire panel of isolated readings. The overall rate of CD/HT-free survival after 12 months was 60% +/- 5%. Patients who were clinically stable with QRS widening and pVO2 decrease values of <10% had a better CD/HT event-free survival rate at 1 year (92% +/- 5% vs 50% +/- 6%; P <.001). CONCLUSIONS: This study indicates that analysis of time-related changes in prognostic parameters provides relevant incremental prognostic information and may help in the risk stratification of patients with HF and the selection of candidates for HT. In particular, patients who were clinically stable and had QRS widening and a pVO2 decreases <10% in a period > or =6 months appear to be characterized by a good prognosis and may not be suitable candidates for HT. PMID- 12891200 TI - Enoxaparin versus tinzaparin in non-ST-segment elevation acute coronary syndromes: the EVET trial. AB - BACKGROUND: Low-molecular weight heparins have different pharmacokinetic and pharmacodynamic characteristics and may vary in efficacy. We compared the efficacy of enoxaparin with that of tinzaparin in the management of non-ST segment elevation acute coronary syndromes (NSTACS). METHODS: A total of 438 patients with NSTACS were randomized to receive subcutaneous treatment with enoxaparin, 100 IU/kg twice daily (equivalent to 1 mg/kg twice daily; n = 220), or tinzaparin, 175 IU/kg once daily, (n = 218) for as long as 7 days. The primary composite end point was recurrent angina, myocardial infarction (or reinfarction), or death at day 7. Secondary end points were the primary end point at day 30 and the occurrence of individual events at days 7 and 30. RESULTS: The incidence of the primary end point was 12.3% in the enoxaparin group and 21.1% in the tinzaparin group (P =.015). At day 7, the rate of recurrent angina was lower with enoxaparin than with tinzaparin (11.8% vs 19.3%). At day 30, the incidences of the composite end point, recurrent angina, and myocardial infarction were also lower with enoxaparin, 17.7% vs 28.0% (P =.012), 17.3% vs 26.1% and 0.5% vs 2.8%, respectively. The rate of revascularization was lower in the enoxaparin group, 8.6% vs 17.9% (P =.010) at day 7 and 16.4% vs 26.1% (P =.019) at day 30. Rates of bleeding complications were similar in the 2 treatment groups. CONCLUSIONS: This study indicates a benefit of enoxaparin (100 IU/kg twice daily) as compared with tinzaparin (175 IU/kg once daily) in the treatment of patients with NSTACS, which is sustained for at least 30 days. PMID- 12891201 TI - Risk factors for "major" embolic events in hospitalized patients with infective endocarditis. AB - BACKGROUND: Infective endocarditis often is complicated by embolic events after hospital admission. Identifying patients at higher risk may improve the disease outcome. This study was aimed at identifying predictors of embolic risk among the clinical and laboratory data obtained on hospital admission in patients diagnosed as having definite infective endocarditis according to the Duke criteria. METHODS: Ninety-four patients were enrolled in a prospective study. The results of hematologic, echocardiographic, and microbiological investigations were analyzed, using statistical methods as appropriate. Multivariate analysis was applied to variables significantly associated with embolism in univariate analysis. RESULTS: Forty-six percent of patients had a major embolic complication after admission. No association was found between embolism and sex, site of infection, or microorganism involved. Patients with embolism were significantly younger, had larger vegetation, and showed a significantly higher level of serum C-reactive protein and lower albumin concentrations than those without embolism. Young age, larger vegetation size, and high levels of C-reactive protein were the independent variables associated with an increased incidence of embolic events in the multivariate logistic regression analysis. CONCLUSIONS: Our data indicate that patients with infective endocarditis with young age and/or with large vegetation and/or with high serum levels of C-reactive protein are at increased risk of major embolic complications during the in-hospital course of the disease. PMID- 12891202 TI - Reduction in obesity and coronary risk factors after high caloric exercise training in overweight coronary patients. AB - BACKGROUND: The majority of patients with coronary heart disease (CHD) are overweight. However, little weight loss occurs with participation in a standard cardiac rehabilitation (CR) program. METHODS: Fifteen overweight patients (average body mass index of 31.0 kg/m2) with CHD completed a 4-month exercise training program in a CR program. The exercise program consisted primarily of walking long duration (60-90 minutes per session) 5 to 7 days per week at a relatively low intensity of 50% to 60% of peak VO2. Measures of body composition by dual-energy x-ray absorptiometry, body fat distribution by computed tomography, plasma lipid-lipoprotein, glucose and insulin concentrations, and peak VO2 were obtained before and after the exercise intervention. Patients maintained an isocaloric diet throughout the study. RESULTS: Patients had reductions in total body weight (-4.6 kg), fat mass (-3.6 kg), percent body fat ( 2.9%), and waist circumference (-5.6 cm) (all P <.001) while maintaining fat-free mass. Subcutaneous adipose tissue was reduced by 12% (P <.001) and visceral adipose tissue was lowered by 14% (P <.001). There were favorable changes in the lipid-metabolic profile with reductions in triglyceride levels (-23.7%), total cholesterol/HDL-C ratio (-14.3%), and fasting insulin levels (-22.3%) (all P <.05). Peak VO2 increased by 21.2% (P <.001). CONCLUSIONS: The present pilot study results suggest that a high caloric training exercise training program in the CR setting may be effective in promoting weight loss and improving coronary risk factors in overweight coronary patients. Although additional research with randomized control patients is needed, this alternative to traditional CR may be considered to maximize weight loss as part of a secondary prevention program. PMID- 12891203 TI - Joint effects of Chlamydia pneumoniae infection and classic coronary risk factors on risk of acute myocardial infarction. AB - BACKGROUND: Although not in itself strongly predictive of coronary heart disease, Chlamydia pneumoniae infection could interact with classic risk factors in determining risk of acute myocardial infarction (AMI). METHODS: We assessed C pneumoniae immunoglobulin (Ig) G and IgA titers and classic risk factors in 618 patients with AMI and in 967 controls. RESULTS: IgG titers were not related to AMI, but a significant association was seen between IgA titers and AMI. Excess risk of AMI was noted mainly among patients with the highest IgA titers, such as those beyond 2.88 (the 95th percentile cutoff point in control subjects), showing a 1.8-fold increase in risk (odds ratio 1.75, 95% CI 1.04-2.92). Classic risk factors did not differ between subjects with IgA titers above and below the 95th percentile cutoff. However, in multivariate analyses, models incorporating both IgA titers and a classic risk factor such as obesity, hypercholesterolemia, or smoking predicted risk more effectively than single-parameter models. For example, the odds ratio for AMI among subjects with the highest IgA titers plus hypercholesterolemia was greater than the product of individual risks associated with these high IgA titers and with hypercholesterolemia. CONCLUSIONS: Interactions with classic risk factors (ie, obesity, hypercholesterolemia, and smoking), increased the predictive value of C pneumoniae IgA antibody titers in determining risk of AMI. PMID- 12891204 TI - Risk factors for myocardial infarction in Brazil. AB - BACKGROUND: Approximately three-quarters of cardiovascular disease deaths in the world come from developing countries, and acute myocardial infarction (AMI) is an important cause of death. Brazil is one of the largest countries in Latin America and the contemporary evaluation of risk factors for AMI is crucial for a more efficacious disease management. METHODS: The Acute Myocardial Infarction Risk Factor Assessment in Brazil (AFIRMAR) study is a case-control, hospital-based study involving 104 hospitals in 51 cities in Brazil, designed to evaluate risk factors for a first ST-segment elevation AMI. RESULTS: A total of 1279 pairs, matched by age (+/- 5 years) and sex, were enrolled. The conditional multivariable analysis of 33 variables showed the following independent risk factors for AMI: > or =5 cigarettes per day (odds ratio [OR] 4.90, P <.00001); glucose > or =126 mg/dL (OR 2.82, P <.00001); waist/hip ratio > or =0.94 (OR 2.45, P <.00001); family history of CAD (OR 2.29, P <.00001), low-density lipoprotein-cholesterol 100 to 120 mg/dL (OR 2.10, P <.00001); reported hypertension (OR 2.09, P <.00001); <5 cigarettes per day (OR 2.07, P =.0171); low density lipoprotein-cholesterol >120 mg/dL (OR 1.75, P <.00001); reported diabetes mellitus (OR 1.70, P =.0069); waist/hip ratio 0.90 to 0.93 (OR 1.52, P =.0212); alcohol intake (up to 2 days/week) (OR 0.75, P <.0309); alcohol intake (3-7 days/week) (OR 0.60, P =.0085); family income R$600 to R$1200 and college education (OR 2.92, P =.0499); family income >R$1200 and college education (OR 0.68, P = 0.0239) CONCLUSIONS: The independent risk factors for AMI in Brazil showed a conventional distribution pattern (smoking, diabetes mellitus and central obesity among others) with different strengths of association; most of them being preventable by implementation of adequate policies. PMID- 12891205 TI - Endocardial pacemaker or defibrillator leads with infected vegetations: a single center experience and consequences of transvenous extraction. AB - BACKGROUND: Removal of infected endovascular leads if often required for cure of systemic infection, but the perceived risk of embolic events in the presence of large (>10 mm) vegetations has been considered a relative contraindication to transvenous removal. Surgical removal of pacemaker leads has been suggested in this situation to avoid occurrence of pulmonary embolization. METHODS: Of 38 patients with infection of implanted pacemaker or cardioverter-defibrillator devices, those with evidence for systemic infection underwent transesophageal echocardiography to assess for the presence of vegetations. RESULTS: Vegetations on endocardial leads or right-sided cardiac structures ranging in size from 10 mm to 38 mm in their largest dimension were detected in 9 patients. All patients underwent successful transvenous removal of endocardial leads. Five of 9 patients (55%) had evidence of pulmonary embolism. However, all 5 patients made a full recovery with antibiotic treatment and anticoagulation. Among patients with endocardial vegetations, there was no difference in hospitalization periods between those with or without pulmonary embolism (14.6 +/- 0.8 days vs 18.0 +/- 4.5 days, P =.7). CONCLUSIONS: Transvenous removal of infected pacemaker leads is an alternative to open-thoracotomy removal of infected leads. Fifty-five percent of patients with vegetations on endocardial leads in our series experienced pulmonary embolism, but neither survival nor length of hospital stay were affected by this complication. PMID- 12891206 TI - Safety of abciximab in patients with chronic renal insufficiency who are undergoing percutaneous coronary interventions. AB - BACKGROUND: Patients with chronic renal insufficiency (CRI) have worse outcomes during and after percutaneous coronary interventions (PCI). Abciximab reduces complications, but may cause excessive bleeding in patients with CRI. Therefore, we sought to determine the safety of abciximab in patients with CRI. METHODS: Patients (n = 4158) undergoing PCI at the Mayo Clinic since abciximab became available were analyzed according to their estimated creatinine clearance (> or =70, 50-69, or <50 mL/min) or need for dialysis. Major bleeding was defined as a cerebrovascular bleed or a decrease in the hematocrit level >15%. Minor bleeding was defined as a decrease in the hematocrit level of 10% to 15% with an identifiable site of bleeding. RESULTS: CRI was associated with increased bleeding in patients who received abciximab and patients who did not. However, there was only a trend toward an interaction between creatinine clearance and major bleeding with abciximab (odds ratio [OR], 1.18; P =.06) and no interaction with minor bleeding (OR, 1.01; P =.94) or any bleeding (OR, 1.10; P =.15). CONCLUSION: CRI is associated with an increased risk of bleeding complications after PCI. Although abciximab increases the risk of bleeding in all patients, the increase in relative risk is not significantly greater in patients with CRI. Thus, abciximab may be given safely in patients with CRI who are undergoing PCI. PMID- 12891207 TI - Effect of fasting glucose levels on mortality rate in patients with and without diabetes mellitus and coronary artery disease undergoing percutaneous coronary intervention. AB - BACKGROUND: Diabetes mellitus (DM) is predictive of increased mortality for patients with coronary artery disease (CAD). To what extent this risk extends below the diabetic threshold (fasting glucose level [FG] <126 mg/dL) is uncertain. METHODS: The study objective was to determine the risk associated with FG in a prospectively assembled cohort of 1612 patients with CAD who were undergoing percutaneous coronary intervention (PCI) and had a FG measured or a clinical diagnosis of DM (CDM). Patients were grouped as: CDM; no CDM, but FG > or =126 mg/dL (ADA-DM); impaired FG, 110-125 mg/dL (IFG); or normal FG, <110 mg/dL (NFG). Survival was assessed for 2.8 +/- 1.2 years. RESULTS: The average patient age was 62 +/- 12 years; 74% of the patients were men. Diagnostic frequencies were: CDM, 24%; ADA-DM, 18%; IFG, 19%; and NFG, 39%. Mortality rates were greater for patients in the CDM (44/394 [11.2%], P <.0001), ADA-DM (27/283 [9.5%], P <.001), and IFG (20/305 [6.6%], P =.04) groups than patients in the NFG group(12/630 [1.9%]). Independent receiver operating characteristic analysis chose FG > or =109 mg/dL as the best cutoff for increased risk (sensitivity, 81%; specificity, 51%). After adjustment with Cox regression analysis, CDM (hazard ratio [HR] = 5.0; 95% CI, 2.6-9.6; P <.001), ADA-DM (HR, 4.1; 95% CI, 2.1-8.2; P <.001), and IFG status (HR, 3.2; 95% CI, 1.5-6.5; P =.002) remained independent predictors of mortality. CONCLUSIONS: Prognostically significant abnormalities of FG are much more prevalent (61%) than expected in patients with CAD who are undergoing PCI. Despite revascularization, the associated mortality risk of even mild elevations in FG is substantial, emphasizing the importance of early detection and treatment of glycemia-related risk. PMID- 12891208 TI - Quantitative clinical assessment of chronic anterior myocardial infarction with delayed enhancement magnetic resonance imaging and QRS scoring. AB - BACKGROUND: Both the regional and global myocardial extent of chronic myocardial infarction (MI) are important prognostic factors for length and quality of life and also crucial for the choice of therapy in patients with ischemic heart disease. Our aim was to develop and validate techniques for comparison between regional and global size of remote anterior MI in the left ventricle quantified with both magnetic resonance imaging (MRI) and electrocardiogram (ECG). METHODS: Delayed-enhancement (DE) MRI was used as a clinical "gold standard" for MI size to evaluate the extent of MI estimated with the commonly available standard 12 lead ECG. A method for comparing global and regional quantifications of MI with DE-MRI and ECG was developed. The Selvester QRS-scoring system was used for estimating MI size electrocardiographically. RESULTS: Twenty-five patients with chronic single anterior MI, documented with DE-MRI, were studied. The best agreement for mean % MI per regional segment of the left ventricle was found in the middle third (26% vs 27%), whereas the most significant discrepancy was found in the apex (56% vs 30%). The global MI size of the left ventricle averaged 21 +/ 9% with DE-MRI and 22% +/- 12% with ECG, with a correlation of r = 0.40 (P <.05). CONCLUSIONS: The current Selvester QRS scoring system performs well for quantifying anterior MI in the mid-regions of the left ventricle. The diagnostic performance of the Selvester QRS-scoring system for quantifying MI in the other regions, particularly the left ventricular apex, can potentially be improved, with DE-MRI as the gold standard. PMID- 12891209 TI - Vocal cord paralysis after transcatheter coil embolization of patent ductus arteriosus. AB - BACKGROUND: In patients with patent ductus arteriosus (PDA) after Gianturco coil embolization, vocal cord paralysis (VCP) had not been previously described. This study investigates the risk factors of coil embolization associated with VCP. METHODS: We reviewed the medical records of all patients who had undergone transcatheter closure of PDA with a Gianturco coil between March 1998 and May 2001, and 75 patients (age range, 6 months to 55 years; mean age, 5.5 years) were identified. The procedure was performed with local anesthesia and with sedation in some young children. No patient required general anesthesia or endotracheal intubation. RESULTS: Three of the 75 patients had hoarseness caused by VCP after coil embolization. Patients with VCP had a longer ductus length (P <.01) and a smaller ductus diameter (P <.01) than patients without VCP. The pathogenesis of VCP may be caused by tense stretching and angulation of the ductus arteriosus induced by the implanted coil, which leads to compression injury of the adjacent left recurrent laryngeal nerve. CONCLUSIONS: A long ductus length (> or =12 mm) and a small ductus diameter (<1 mm) are significant risk factors of VCP after coil embolization in children <1 year of age. We recommend that coil embolization be performed very carefully in patients with PDA. PMID- 12891210 TI - Prognostic significance of morphometric endomyocardial biopsy analysis in patients with idiopathic dilated cardiomyopathy. AB - BACKGROUND: To date, considerable controversy exists on the prognostic significance of morphometric endomyocardial biopsy findings in patients with idiopathic dilated cardiomyopathy (IDC). METHODS: Quantitative analyses of interstitial structured tissue, myofibril volume fraction, and myocytic fiber diameters of left ventricular endomyocardial biopsy specimens were performed in 124 patients with IDC. RESULTS: During 51 +/- 22 months follow-up after left ventricular endomyocardial biopsy, major arrhythmic events, defined as sustained ventricular tachycardia (VT), ventricular fibrillation (VF), or sudden cardiac death, were observed in 24 patients (19%). Death from any cause or heart transplant was observed in 39 patients (31%). The amount of interstitial structured tissue, myofibril volume fraction, and myocytic fiber diameters determined from left ventricular endomyocardial biopsy specimens did not differ significantly between patients with and patients without major arrhythmic events or between patients with and patients without transplant-free survival during follow-up. CONCLUSIONS: Quantitative analysis of the amount of interstitial structured tissue, myofibril volume fraction, and myocytic fiber diameters in left ventricular endomyocardial biopsy specimens does not appear to be useful for predicting arrhythmic events and transplant-free survival in IDC. PMID- 12891211 TI - Effect of n-3 fatty acids on heart rate variability and baroreflex sensitivity in middle-aged subjects. AB - BACKGROUND: N-3 fatty acids may reduce the risk of sudden death by preventing life-threatening cardiac arrhythmias. Heart rate variability (HRV) and baroreflex sensitivity (BRS) reflect cardiac autonomic regulation; reduced values predict arrhythmic events and mortality. Effects of n-3 fatty acids on these risk indicators of arrhythmia have not been widely studied. We investigated the effect of supplemental intake of n-3 fatty acids on HRV and BRS in apparently healthy subjects aged 50 to 70 years. METHODS: After a run-in period of 4 weeks, 84 subjects were randomized to receive capsules with either 3.5 g of fish oil or placebo oil daily for 12 weeks. Before and after intervention, electrocardiograms and blood pressure were recorded for 10 minutes with standardized respiration of 15 breaths/min. The SD of the duration of all normal RR intervals (SDNN10) and the root mean square successive differences (RMSSD10) were calculated from the 10 minute recordings. We also computed low frequency power (LF) and high frequency power (HF). BRS was computed by integrating the spectral systolic blood pressure to interbeat-interval transfer function in the 0.05- to 0.15-Hz band. RESULTS: The different measures of HRV and BRS did not significantly improve with the intake of n-3 fatty acids. SDNN10 decreased by 3.05 ms or 7.7% (95% CI, -8.91 2.82 ms), and BRS decreased by 0.92 ms/mm Hg or 0.1% (95% CI, -2.66-0.81 ms/mm Hg) in the fish oil group compared with the placebo group. CONCLUSIONS: N-3 fatty acids have no effect on HRV from short-term recordings and BRS in apparently healthy subjects. PMID- 12891212 TI - Pioglitazone reduces neointimal tissue proliferation after coronary stent implantation in patients with type 2 diabetes mellitus: an intravascular ultrasound scanning study. AB - BACKGROUND: It has been reported that pioglitazone reduces neointimal hyperplasia after balloon-induced vascular injury in an experimental model. METHODS: To determine whether pioglitazone reduces neointimal tissue proliferation after coronary stent implantation in patients with type 2 diabetes mellitus, we studied 44 stented lesions in 44 patients with diabetes mellitus who underwent successful coronary stent implantation. Study patients were randomized into 2 groups: the pioglitazone group (23 patients with 23 lesions) and the control group (21 patients with 21 lesions). All patients underwent serial quantitative coronary angiography and serial intravascular ultrasound scanning studies. With a motorized pullback system, multiple image slices within the stent were obtained at every 1 mm. The stent area and lumen area were measured, and the neointimal area was calculated. Measurements were averaged over the number of selected image slices. The neointimal index was calculated as the averaged neointimal area divided by the averaged stent area multiplied by 100 (%). RESULTS: After 6 months of treatment, angiographic in-stent restenosis (17% vs 43%, respectively, P =.0994) and target lesion revascularization (13% vs 38%, respectively, P =.0835) were less frequent in the pioglitazone group than the control group; however, these differences did not reach significance. The intravascular ultrasound scanning study demonstrated that the neointimal index in the pioglitazone group was significantly smaller than that in the control group (28% +/- 9% vs 48% +/- 15%, respectively, P <.0001). CONCLUSION: A serial intravascular ultrasound scanning assessment demonstrated that pioglitazone reduces neointimal tissue proliferation after coronary stent implantation in patients with type 2 diabetes mellitus. PMID- 12891213 TI - Diabetes mellitus and acute myocardial infarction: more data supporting a poorer microvasculature reperfusion. PMID- 12891214 TI - Don't shoot the messenger: the 6-minute walk test is a useful outcome measure in exercise trials. PMID- 12891217 TI - A primer for nuclear terrorism. AB - Mass exposure to radiologic substances presents a unique challenge to the entire response effort, which includes health care professionals, law enforcement personnel, and other first responders. Recognition of signs and symptoms of exposure, and focus on removal and decontamination are priorities of management. Radiation injuries require specialized equipment and access to experts. Patients can have complex patterns of injury, ranging from trauma and the immediate results of an explosion or exposure, to progressive damage associated with radiation sickness. Both conventional injury and radiation illness may require critical care management. Remembering the essentials of first response, that is, treat the patient, not the poison, by addressing the ABCs of airway, breathing, and circulation, is critical to appropriate treatment of radiation exposure. Understanding the basic science of radiologic agents will aid the provider in managing affected patients and preventing further casualties. PMID- 12891219 TI - Clinical Pharmacology & Therapeutics annual report 2002. PMID- 12891220 TI - Side effects of opioids during short-term administration: effect of age, gender, and race. AB - OBJECTIVE: Little is known about risk factors that increase the risk of development of opioid side effects. Our objective was to evaluate the effect of the type of opioid, age, gender, and race on the incidence of side effects from short-term opioid use. METHODS: A secondary analysis of a retrospective cohort study in 35 community-based and tertiary hospitals was done. There were 8855 black or white subjects aged 16 years and older. Patients received meperidine (INN, pethidine), morphine, or fentanyl as part of their treatment. Measurements were made to assess the presence of nausea and vomiting and respiratory depression. RESULTS: Of the patients, 26% had nausea and vomiting and 1.5% had respiratory depression after opioid administration. After adjustment for opioid dose, route of administration, age, gender, and race, meperidine produced less nausea and vomiting (odds ratio [OR] = 0.7; 95% confidence interval [CI], 0.5 0.8) and less respiratory depression (OR = 0.6; 95% CI, 0.2-0.9) than morphine. The risk of respiratory depression increased with age. Compared with patients aged between 16 and 45 years, those aged between 61 and 70 years had 2.8 times the risk of development of respiratory depression (95% CI, 1.2-6.6); those aged between 71 and 80 years had 5.4 times the risk (95% CI, 2.4-11.8); and those aged older than 80 years had 8.7 times the risk (95% CI, 3.8-20.0). Men had less nausea and vomiting than women (OR = 0.5; 95% CI, 0.4-0.6). White subjects had more nausea and vomiting than black subjects (OR = 1.4; 95% CI, 1.1-1.7). CONCLUSIONS: Meperidine produced fewer side effects than morphine during short term use. The risk of respiratory depression increases substantially after 60 years of age. Women have nausea and vomiting more often than men. The effect of race deserves further investigation. PMID- 12891221 TI - Peripheral and central antihyperalgesic effects of diclofenac in a model of human inflammatory pain. AB - AIM: Experimental evidence suggests that the antihyperalgesic effect of nonsteroidal anti-inflammatory drugs may include both peripheral (inflammatory site) and central sites of action. The aim of this study was to assess peripheral and central antihyperalgesic effects of diclofenac in a human experimental pain model. METHODS: This was a randomized, double-blind, placebo-controlled study designed to compare the antihyperalgesic efficacy of topical (65 mg) and oral (93 mg) diclofenac via estimates of mechanical pain thresholds obtained at the site of induced inflammation with von Frey hairs. The dose of the 2 diclofenac formulations was calculated to achieve similar target concentrations, monitored in the inflammatory tissue by intradermal microdialysis. Simultaneous serial blood samples were collected to determine the systemic concentrations of the drug. RESULTS: Diclofenac was superior to placebo 1 hour after topical application (P <.002) and 1 to 3 hours after oral intake (P <.016). Topical diclofenac was more effective than oral diclofenac 1 hour after dosing and produced higher tissue concentrations (46.1 ng/mL versus 11.4 ng/mL, P <.02), whereas the compound was not detectable in plasma. Oral diclofenac had a higher antihyperalgesic efficacy at later observation periods (2-2.5 hours after dosing), when tissue concentrations of diclofenac for the 2 treatments did not differ significantly. The overall pain relief over a 3-hour postdose period was 1.7-fold greater with oral diclofenac than with topical diclofenac. However, the total tissue area under the curve after oral diclofenac did not exceed that for the topical formulation (32.2 ng. h. mL(-1) versus 40.7 ng. h. mL(-1)). CONCLUSION: The higher antihyperalgesic efficacy of oral diclofenac as compared with topical diclofenac at comparable tissue concentrations suggests that not only peripheral but also central mechanisms are involved in the antihyperalgesic effects of systemically administered diclofenac. PMID- 12891222 TI - Time course of recovery of cytochrome p450 3A function after single doses of grapefruit juice. AB - BACKGROUND: Components of grapefruit juice may impair the activity of intestinal cytochrome P450 (CYP) 3A enzymes, sometimes resulting in clinically important drug interactions. The time course of recovery from CYP3A inhibition after a single exposure to grapefruit juice is not clearly established. METHODS: Healthy volunteer subjects (N = 25) received a single 6-mg oral dose of the CYP3A substrate midazolam in the control condition without exposure to grapefruit juice. Two days later, midazolam was administered 2 hours after 300 mL of regular strength grapefruit juice. Subjects were then randomly assigned to 3 different groups, receiving a third midazolam challenge at 26, 50, or 74 hours after exposure to grapefruit juice. The capacity of 6'7'-dihydroxybergamottin and bergamottin to inhibit human CYP3A was studied in vitro using human liver microsomes. RESULTS: The area under the plasma concentration curve (AUC) for midazolam increased by a factor of 1.65 (ratio compared with control) when midazolam was given 2 hours after grapefruit juice. At 26, 50, and 74 hours after grapefruit juice, the AUC ratios (mean AUC value at the indicated time divided by the mean control AUC on day 1) were 1.29, 1.29, and 1.06, respectively. The relationship of time after grapefruit juice exposure versus AUC increase over control indicated a recovery half-life estimated at 23 hours. The midazolam elimination half-life did not change significantly from the control value at any time after grapefruit juice exposure. 6'7'-Dihydroxybergamottin inhibited midazolam alpha-hydroxylation in vitro, with a mean 50% inhibitory concentration of 4.7 micro mol/L; preincubation of microsomes with 6'7'-dihydroxybergamottin greatly reduced the 50% inhibitory concentration to 0.31 micro mol/L, consistent with mechanism-based inhibition. Bergamottin itself had much weaker inhibitory potency compared to 6'7'-dihydroxybergamottin. CONCLUSIONS: A usual single exposure to grapefruit juice appears to impair the enteric, but not the hepatic, component of presystemic extraction of oral midazolam. Recovery is largely complete within 3 days, consistent with enzyme regeneration after mechanism-based inhibition. 6'7'-Dihydroxybergamottin was verified as a potent mechanism-based inhibitor of midazolam alpha-hydroxylation by CYP3A in vitro. PMID- 12891223 TI - Celecoxib inhibits metabolism of cytochrome P450 2D6 substrate metoprolol in humans. AB - OBJECTIVE: In vitro data have shown that celecoxib inhibits the metabolism of cytochrome P450 (CYP) 2D6 substrates. However, very limited data are available on the influence of cyclooxygenase 2 inhibitors on the disposition of CYP2D6 substrates in humans. Therefore the objective of this study was to examine the effect of celecoxib and rofecoxib on the pharmacokinetics of the clinically relevant CYP2D6 substrate metoprolol. METHODS: An open, randomized, 3-period crossover study was performed in 12 healthy male volunteers. Metoprolol (50 mg) was given in all 3 periods without or after 7 days of pretreatment with celecoxib (200 mg twice daily) or rofecoxib (25 mg daily) to achieve steady-state conditions of cyclooxygenase 2 inhibitors in periods 2 and 3. RESULTS: Celecoxib significantly increased the area under the plasma concentration-time curve of metoprolol from 271 to 414 micro g. h/L (64% +/- 57%, P <.001) and by more than 200% in 1 volunteer. The extent of this drug interaction was more pronounced in volunteers with 2 fully functional alleles compared with volunteers with 1 fully functional allele (103% +/- 75% versus 36% +/- 23%, P <.05). After administration of celecoxib, the area under the plasma concentration-time curve from 0 to 24 hours of alpha-hydroxymetoprolol decreased significantly from 474 to 387 micro g. h/L (P <.01). Rofecoxib caused no significant effects on the pharmacokinetics of metoprolol. CONCLUSION: We conclude that celecoxib inhibits the metabolism of the CYP2D6 substrate metoprolol but that rofecoxib does not. Clinically relevant drug interaction may occur between celecoxib and CYP2D6 substrates, particularly those with a narrow therapeutic index. PMID- 12891224 TI - Pharmacokinetic drug interactions in children taking oxcarbazepine. AB - OBJECTIVE: Our objective was to evaluate the drug-drug interactions of oxcarbazepine with coadministered antiepileptic drugs in children. METHODS: In a clinical trial, pediatric patients receiving an oxcarbazepine dose titrated to 30 to 46 mg. kg(-1). d(-1) given twice daily had 1 to 4 blood samples collected per patient for population pharmacokinetic analysis of oxcarbazepine's major bioactive 10-monohydroxy metabolite. With the use of NONMEM, 7 concomitant antiepileptic drugs and 12 additional covariates were examined for their effects on the pharmacokinetics of 10-monohydroxy metabolite. In addition, for each concomitant antiepileptic drug, the ratio of its mean concentration with coadministration of oxcarbazepine to that without coadministration at baseline was calculated to evaluate the effect of oxcarbazepine on the coadministered antiepileptic drugs. RESULTS: The population pharmacokinetic data for 10 monohydroxy metabolite consisted of a total of 376 observations from 109 patients, aged 3 to 17 years. Body surface area and 3 antiepileptic drugs (carbamazepine, phenobarbital, and phenytoin) were significant predictors of the apparent clearance of 10-monohydroxy metabolite, whereas height was a significant predictor of apparent volume. Weight-normalized clearance of 10-monohydroxy metabolite was higher in young children than in older children and adults. Carbamazepine, phenobarbital, or phenytoin administered with oxcarbazepine increased the apparent clearance of 10-monohydroxy metabolite by 31% to 35%, whereas carbamazepine levels decreased by 15% and phenobarbital levels increased by 14%. CONCLUSIONS: Oxcarbazepine has a low propensity to inhibit or induce oxidative enzymes. Young children could be given higher milligrams-per-kilogram oxcarbazepine doses than older children and adults to achieve the same mean steady-state concentration of 10-monohydroxy metabolite. The adjustment is based simply on body size. PMID- 12891225 TI - Effects of aprepitant on cytochrome P450 3A4 activity using midazolam as a probe. AB - BACKGROUND: Aprepitant is a neurokinin(1) receptor antagonist that enhances prevention of chemotherapy-induced nausea and vomiting when added to conventional therapy with a corticosteroid and a 5-hydroxytryptamine(3) (5-HT(3)) antagonist. Because aprepitant may be used with a variety of chemotherapeutic agents and ancillary support drugs, which may be substrates of cytochrome P450 (CYP) 3A4, assessment of the potential of this drug to inhibit CYP3A4 activity in vivo is important. The effect of aprepitant on in vivo CYP3A4 activity in humans with oral midazolam used as a sensitive probe of CYP3A4 activity was evaluated in this study. METHODS: In this open-label, randomized, single-period study, 16 healthy male subjects were enrolled. Subjects received one of two oral aprepitant regimens for 5 days (8 subjects per regimen): (1) 125 mg aprepitant on day 1 and then 80 mg/d on days 2 to 5 or (2) 40 mg aprepitant on day 1 and then 25 mg/d on days 2 to 5. All subjects also received a single oral dose of midazolam, 2 mg, at prestudy (3 to 7 days before aprepitant treatment) and on days 1 and 5 (1 hour after aprepitant administration). RESULTS: Coadministration of midazolam and 125/80 mg aprepitant increased the midazolam area under the plasma concentration time curve by 2.3-fold on day 1 (P <.01) and by 3.3-fold on day 5 (P <.01), as compared with midazolam alone (prestudy). The 125/80-mg regimen of aprepitant also increased the midazolam maximum observed concentration by 1.5-fold on day 1 (P <.05) and by 1.9-fold on day 5 (P <.01). The midazolam half-life values increased from 1.7 hours (prestudy) to 3.3 hours on both day 1 and day 5. Coadministration of 40/25 mg aprepitant and midazolam did not result in significant changes in the midazolam area under the plasma concentration-time curve, maximum observed concentration, and half-life at either day 1 or day 5. CONCLUSIONS: The 5-day 125/80-mg regimen of aprepitant produced moderate inhibition of CYP3A4 activity in humans, as measured with the use of midazolam as a probe drug. PMID- 12891226 TI - Pharmacokinetics and preliminary clinical data of the novel chemoprotectant BNP7787 and cisplatin and their metabolites. AB - INTRODUCTION: BNP7787 (disodium 2,2'-dithio-bis-ethane sulfonate) is currently undergoing development as a chemoprotective agent to prevent common and serious cisplatin-induced side effects. In the kidneys, intestine, and liver, BNP7787 is believed to undergo intracellular conversion into 2-mercaptoethane sulfonate (mesna), which can locally inactivate toxic platinum species. Methods and objectives In a phase I trial, 25 patients with advanced solid tumors received a 1-hour intravenous infusion of 75 mg/m(2) cisplatin immediately preceded by a 15 minute intravenous infusion of BNP7787 every 3 weeks. For pharmacokinetic investigation of BNP7787 and mesna and a possible mutual pharmacokinetic interaction between BNP7787 and cisplatin, cisplatin and BNP7787 were also administered as single agents in 14 of 25 patients. The dose of BNP7787 was escalated from 4.1 to 41 g/m(2). Patients were also monitored for tumor response and possible side effects from BNP7787. RESULTS: The maximum plasma concentration of mesna was reached approximately 1.7 hours after the start of the BNP7787 infusion. The maximum plasma concentration and area under the curve to infinity (AUC( infinity )) of BNP7787 and mesna increased linearly with the dose. The mean volume of distribution of BNP7787 (+/-SD) was approximately 0.26 +/- 0.08 L/kg. The mean normalized AUC( infinity ) of mesna was only approximately 8% of the normalized AUC( infinity ) of BNP7787. The pharmacokinetic profile of mesna was unaffected by cisplatin and its metabolites. None of the dose levels of BNP7787 (4.1-41 g/m(2)) administered appeared to influence the pharmacokinetic profile of total platinum, unbound platinum, or monohydrated cisplatin. The observed effects regarding a possible mutual interaction between BNP7787 and intact cisplatin were minor, and none were statistically significant at BNP7787 dose levels of 18.4 to 41 g/m(2). The confidence intervals for the pharmacokinetic parameters of BNP7787 and intact cisplatin, however, were relatively broad. Overall, BNP7787 was well tolerated at all dose levels (4.1-41.0 g/m(2)). The most frequently reported event related to BNP7787 was local intravenous site discomfort; the majority of events were mild (grade 1). Side effects of BNP7787 at the highest dose level of 41 g/m(2) were more prominent and included nausea and vomiting, as well as a warm feeling or flushing (grade 2 or lower). Partial tumor responses and stable disease were measured in 12 of 25 patients. CONCLUSION: BNP7787 was relatively nontoxic at doses up to 41 g/m(2). The combination of BNP7787 with cisplatin did not alter the pharmacokinetic profiles of mesna or the cisplatin metabolites. At the higher dose levels of BNP7787 (18.4 to 41 g/m(2)), there appeared to be no mutual interaction between BNP7787 and intact cisplatin, which needs to be confirmed in a larger number of patients. The absence of a mutual interaction between BNP7787 and intact cisplatin is consistent with the observation that several patients had objective tumor responses with BNP7787 and cisplatin administration. PMID- 12891227 TI - Effects of garlic (Allium sativum L.) supplementation on cytochrome P450 2D6 and 3A4 activity in healthy volunteers. AB - Garlic (Allium sativum L.) is a commonly used food and herbal supplement. The objective of this study was to assess in healthy volunteers (N = 14) the influence of a garlic extract on the activity of cytochrome P450 (CYP) 2D6 and 3A4. Probe substrates dextromethorphan (CYP2D6) and alprazolam (CYP3A4) were administered orally at baseline and again after treatment with garlic extract (3 x 600 mg twice daily) for 14 days. Urinary dextromethorphan/dextrorphan ratios and alprazolam plasma concentrations were determined by HPLC at baseline and after garlic extract treatment. The ratio of dextromethorphan to its metabolite was 0.044 +/- 0.48 at baseline and 0.052 +/- 0.095 after garlic supplementation. There were no significant differences between the baseline and garlic phases (P > or =.05). For alprazolam, there were no significant differences in pharmacokinetic parameters at baseline and after garlic extract treatment (all P values > or =.05; maximum concentration in plasma, 27.3 +/- 2.6 ng/mL versus 27.3 +/- 4.8 ng/mL; time to reach maximum concentration in plasma, 1.9 +/- 1.4 h versus 2.4 +/- 1.8 h; area under the time-versus-concentration curve, 537 +/- 94 h. ng. mL(-1) versus 548 +/- 159 h. ng. mL(-1); half-life of elimination, 13.7 +/ 4.4 h versus 14.5 +/- 4.3 h). Our results indicate that garlic extracts are unlikely to alter the disposition of coadministered medications primarily dependent on the CYP2D6 or CYP3A4 pathway of metabolism. PMID- 12891228 TI - Pharmacokinetics and pharmacodynamics of pravastatin in children with familial hypercholesterolemia. AB - BACKGROUND: Pravastatin is a widely used statin in adults, but its pharmacokinetics in children is not known. Our aim was to determine the single dose pharmacokinetics and the lipid-lowering effect and safety of short-term administration of pravastatin in children. METHODS: Twenty children (age range, 4.9-15.6 years) with heterozygous familial hypercholesterolemia ingested a single dose of 10 mg pravastatin. Plasma concentrations of pravastatin were measured for up to 10 hours. The patients then took 10 mg pravastatin orally once daily for 8 weeks. The concentration of serum lipids and safety laboratory parameters were measured before and after 8 weeks of treatment. RESULTS: The mean peak plasma concentration (C(max)) of pravastatin was 15.7 ng/mL (range, 1.6-55.0 ng/mL), and the mean time to reach C(max) was 1.4 hours (range, 0.5-4 hours). The mean elimination half-life of pravastatin was 1.6 hours (range, 0.85-4.2 hours). The area under the plasma concentration-time curve of pravastatin ranged from 5.7 to 58.9 ng. h/mL (mean value, 26.6 ng. h/mL). By 8 weeks of treatment, the serum concentration of total cholesterol had decreased 18% (P <.0001); low-density lipoprotein cholesterol, 21% (P <.0001); and triglycerides, 18% (not significant, P =.18). The concentration of high-density lipoprotein cholesterol had increased 8% (not significant, P =.13). Few transient adverse events occurred. No increases in serum alanine aminotransferase, creatine kinase, or creatinine level were observed. CONCLUSIONS: The pharmacokinetic and pharmacodynamic profile of pravastatin in children is similar to that reported for adults. In the short term, the daily dose of 10 mg pravastatin was well tolerated and moderately effective in decreasing the serum cholesterol concentration. However, further studies are needed on the long-term safety and efficacy of pravastatin in children. PMID- 12891229 TI - Influence of CYP2C9 polymorphisms on the pharmacokinetics and cholesterol lowering activity of (-)-3S,5R-fluvastatin and (+)-3R,5S-fluvastatin in healthy volunteers. AB - INTRODUCTION: In vitro data indicate that biotransformation of the synthetic 3 hydroxy-3-methylglutaryl-coenzyme A reductase inhibitor fluvastatin is catalyzed by the cytochrome P450 (CYP) enzyme 2C9. The consequences of CYP2C9 genetic polymorphisms on fluvastatin pharmacokinetics and on its efficacy have not been investigated in humans thus far. METHODS: Twenty-four healthy heterozygous or homozygous carriers of the CYP2C9 variants Arg144Cys (*2) and Ile359Leu (*3) and 2 individuals with the deficient CYP2D6 genotype *4/*4 took 40 mg racemic fluvastatin daily for 14 days. All subjects had also been genotyped for CYP2C8, CYP2C19, and CYP2D6 polymorphisms. Pharmacokinetics was analyzed after the first fluvastatin administration. Serum lipid concentrations were measured before fluvastatin intake and on day 15. Plasma concentrations of (+)-3R,5S-fluvastatin and of (-)-3S,5R-fluvastatin were quantified by enantiospecific HPLC. RESULTS: Pharmacokinetics of both enantiomers showed statistically significant differences according to the number of CYP2C9*3 alleles (P <.0001, F test). Mean (and SD) values for area under the curve of the active (+)-3R,5S-fluvastatin in carriers of the genotype CYP2C9*1/*1, *1/*3, and *3/*3 were 173 (85) micro g. L(-1). h, 231 (85) micro g. L(-1). h, and 533 (120) micro g. L(-1). h, respectively. The corresponding values for area under the curve of (-)-3S,5R-fluvastatin were 227 (133) micro g. L(-1). h, 360 (103) micro g. L(-1). h, and 1126 (311) micro g. L( 1). h for CYP2C9*1/*1, *1/*3, and *3/*3, respectively. The CYP2C9*2 variant did not have any significant influence on fluvastatin kinetics, nor did the CYP2C8*3 allele, which was tightly linked with CYP2C9*2. Total serum cholesterol and low density lipoprotein cholesterol concentrations decreased significantly during the 14-day treatment period (P <.001), but no correlation with the CYP2C9 genotype was found. CONCLUSIONS: The pharmacokinetics of both enantiomers of fluvastatin depended on the CYP2C9 genotype, with a 3-fold group mean difference in the active enantiomer and even greater differences in the inactive enantiomer, but differences in plasma concentrations were not reflected in cholesterol lowering after 14 days of fluvastatin intake in healthy volunteers. PMID- 12891230 TI - Rational prescribing of essential medicines. PMID- 12891231 TI - MDR1 and CYP3A4 polymorphisms among African, Indian, and white populations in KwaZulu-Natal, South Africa. PMID- 12891232 TI - [Diagnostic value of heart fatty-acid binding protein in early hospitalized patients with non ST elevation acute coronary syndrome]. AB - BACKGROUND: Heart fatty-acid-binding protein (FABP) is supposed to be the most sensitive biomarker of myocardial necrosis in patients with Q-wave myocardial infarction (MI) and non-diagnostic ECG during first hours after onset of symptoms. However, diagnostic value of FABP in patients with non-ST elevation acute coronary syndrome (NSTEACS) is not well established. AIM: To elucidate diagnostic value of FABP in patients with NSTEACS hospitalized within time interval considered to be too early for a majority of biochemical tests. MATERIAL AND METHODS: FABP levels were measured by immunofluorometry (HyTest, Finland) in 44 patients (26 men, mean age 69+/-8.9 years) at admission within 6 hours (median - 2 h) from onset of index attack of angina and in 6, 12, 24 hours after onset of pain. Cut off FABP level was 12 ng/ml. Serum cardiac troponin I was measured for diagnosis of MI on admission and twice during first 24 hours of hospital stay. Cut off TnI level was 0.4 ng/ml. RESULTS: Acute MI was diagnosed by TnI above cut off in 31 patients (70.5%). There were no new-Q-wave MIs. Average ratio of observed serum FABP level to diagnostic cut off value on admission and in 6, 12, 24 hours after onset of pain was higher in patients with MI than in patients with unstable angina (1.01, 1.53, 0.81, 0.66 and 0.78, 0.51, 0.65, 0.56, respectively). The difference was maximally significant in 6 hours after onset of pain (p=0.018). Among patients with MI admission FABP compared with admission TnI more frequently exceeded diagnostic level (in 18 vs 9 patients, respectively, p=0.009). Sensitivity and specificity of admission levels of FABP and TnI for diagnosis of MI were 58 and 85%, 29% and 100%, respectively. CONCLUSION: In patients with NSTEACS during first 6 hours after pain onset FABP compared with TnI has greater sensitivity for detection of MI and sufficient specificity. FABP can be used as additional diagnostic tool for MI detection in early admitted patients with NSTEACS. PMID- 12891233 TI - [Diabetic dyslipidemia and pathogenesis of ischemic heart disease]. AB - BACKGROUND: Type II diabetes is associated with high risk of ischemic heart disease (IHD), diabetes and IHD have similar risk factors, abdominal obesity as well as interrelated disturbances of lipid and carbohydrate metabolism play important roles in pathogenesis of both diseases. AIM: To elucidate characteristics of metabolism and transport of lipids and carbohydrates in patients with type II diabetes and IHD with normal and excessive body mass with abdominal type of fat distribution. MATERIAL AND METHODS: Parameters of carbohydrate metabolism, blood serum levels of free fatty acids, lipid and apoprotein (apo-) parameters of lipoprotein spectrum were evaluated in subjects without diabetes or IHD (group 1), patients with IHD and normal body mass (group 2), patients with IHD combined with diabetes with normal body mass (group 3), patients with IHD, diabetes and abdominal obesity (group 4). RESULTS: Patients with combination of diabetes and IHD compared with those of groups 1 and 2 in addition to higher blood glucose had lower growth hormone, higher triglycerides, lower high density lipoprotein cholesterol (CH) and apo-A1, higher low/high density lipoprotein CH and apo-B/apo-A1 ratios. These atherogenic changes were more pronounced in patients of group 4 who had highest levels of insulin, free fatty acids, low density lipoprotein CH and apo-B and lowest glucose/insulin ratio and level of apo-A1. CONCLUSION: Insulin resistance and hyperinsulinemia are most important features of disturbed carbohydrate and lipid metabolism. Insulin resistance and hyperinsulinemia are most pronounced in subjects with abdominal obesity in whom they create conditions for impaired glucose utilization, development of atherogenic dyslipidemia and eventually emergence of IHD. PMID- 12891234 TI - [Quantitative assessment of right ventricular contractility by contrast angiography in healthy persons and patients with ischemic heart disease]. AB - Absence of special programs for quantitative analysis of right ventricular volumes and function precludes precise evaluation of right ventriculography data. We used standard programs designed for analysis of left ventriculograms for evaluation of right ventricular volumes and function in 32 patients without and 20 patients with coronary artery disease. Left ventriculography and right angiography were carried out in 30- and 60-degree right and left anterior oblique projections, respectively. Average correction factors obtained by comparison of left and right ventricular stroke volumes were 0.9243+/-0.2887 and 0.8758+/ 0.2232 for patients without and with coronary artery disease, respectively. Such conversion was considered to be simple and adequate method of adaptation of existing programs of quantitative analysis of angiograms for evaluation of right ventricular volumes and function. PMID- 12891235 TI - [Medical care of persons aged 25-84 years with acute myocardial infarction in Kaunas in 1996 (ischemic heart disease register data)]. AB - AIM: To evaluate peculiarities of medical care provided for patients with acute myocardial infarction (AMI) depending on sex and age in Kaunas in 1996. METHODS: Kaunas community-based ischemic heart disease register conducted according to the WHO MONICA Project methodology was the source of the data. RESULTS: Number of diagnosed acute myocardial infarctions among Kaunas population aged 25-84 years in 1996 was 1269. Only 28 day AMI survivors were considered in the present study (n=626). During first 4 hours from onset of chest pain male patients sought for medical assistance as frequently as female AMI patients: 38.1% and 34.0%, respectively. A mobile team provided first medical aid in 62.3% of cases. Thrombolytics were used in each 11th men and in 3% of women. Percutaneous coronary angioplasty was carried out in 4.9 and 1.2% of men and women, respectively. Coronary angiography and angioplasty were more often performed in men (13.1 and 4.9%, respectively) than in women (2.7 and 1.2, respectively). PMID- 12891236 TI - [Target organs involvement in arterial hypertension. The role of heredity and environment (study of twins)]. AB - AIM: To elucidate relationship between hereditary and environmental factors in development of target organ damage in hypertension. MATERIAL AND METHODS: The state of myocardium, central nervous system, blood vessels (including funduscopic examination) was assessed in 64 pairs of mono- and dizygous hypertensive twins (mean age 57.2+/-3.2 years) and clinical and genetic analysis of qualitative and quantitative signs was carried out. RESULTS: Hypertensive twins had similar qualitative features of target organs involvement irrespective of duration of hypertension. Concordance of signs of target organ damage in homozygous twins was more than twice higher than that of same signs in dizygous twin pairs. Left ventricular myocardial mass and intima-media thickness were mainly determined by hereditary factors. Their contribution into overall phenotypic variability of a sign exceeded 70%. PMID- 12891237 TI - [Heart rate variability in patients with hypertension and type 2 diabetes treated with long acting calcium antagonists]. AB - BACKGROUND: Enhanced sympathetic activity is a potential cause of increased risk of cardiovascular complications during treatment of hypertension and type 2 diabetes. MATERIAL AND METHODS: Heart rate variability (HRV) was assessed before and after 12-16 weeks of treatment with calcium antagonists in 89 patients with mild and moderate hypertension. RESULTS: Changes of majority of parameters of 24 hour blood pressure monitoring were in general favorable however lowering of magnitude and rate of morning blood pressure rise was observed only in verapamil treated patients. Analysis of HRV showed that treatment with all long acting calcium antagonists under study was associated with enhancement of sympathetic activity. In supine position nifedipine treated compared with verapamil treated patients exhibited greater sympathetic activity. In upright position changes of HRV parameters characteristic for increased sympathetic activity were also more pronounced in dihydropyridine treated compared with verapamil treated patients. CONCLUSION: Effect of calcium antagonists on parameters of HRV is negative and this should be taken into consideration when antihypertensive treatment of patients with hypertension and type 2 diabetes is selected. PMID- 12891238 TI - [Polymorphisms of angiotensin converting enzyme and angiotensin II type 1 receptors genes and the state of regional hemodynamics in borderline arterial hypertension in young men]. AB - Relationship between polymorphisms of ACE and ATR1 genes to morphological and functional states of cardiovascular system was studied in young men with borderline hypertension living in north-west region of Russia. Prevalences of ACE DD and ATR1 AC genotypes were 46 and 54%, respectively. There were no significant correlations between genotypes studied and parameters of central, intracardiac, ophthalmic hemodynamics, as well as presence and severity of albuminuria. Compared with other gene combinations simultaneous carriers of DD and CC genotypes had significantly more pronounced changes of target organs. PMID- 12891239 TI - [Efficacy and safety of simvastatin in patients with hypercholesterolemia (results of a Multicenter Clinical Study)]. AB - AIM: To assess efficacy and safety of a generic preparation of simvastatin Vasilip (KRKA) in ischemic heart disease patients with hypercholesterolemia or combined hyperlipidemia. MATERIAL: Patients (n=167, age 36-73 years) with ischemic heart disease and moderate hypercholesterolemia or combined hyperlipidemia. METHODS: Duration of simvastatin administration was 12 weeks. All patients received 20 mg/day for 6 weeks. Then those patients who did not achieve target low density lipoprotein (LDL) cholesterol level (3.0 mmol/l) were given 40 mg/day for 6 more weeks. Other patients continued to take 20 mg/day. RESULTS: The use of Vasilip was associated with lowering of total, LDL cholesterol and triglycerides (by 28, 39 and 10%, respectively) and 18% elevation of high density lipoprotein cholesterol. Target LDL CH level was achieved in 107 patients (66.9%). In 47 patients (29.4%) LDL CH concentration remained higher than target level but decrease of LDL CH exceeded 10%. Six patients (3.8%) did not respond to therapy. Treatment was stopped because of adverse effects in 2.4% of patients. CONCLUSION: A generic preparation of simvastatin Vasilip in a 12-week open noncomparative study demonstrated substantial lipid lowering activity and did not induce serious adverse reactions. PMID- 12891240 TI - [Hemodynamics, electrophysiological parameters of the heart and differential therapy of attacks of atrial fibrillation in patients with subclinical thyroid dysfunction]. AB - Thirty patients with subclinical thyrotoxicosis and 32 with subclinical hypothyroidism with attacks of atrial fibrillation were investigated. At levels of thyroid-stimulating hormone below 0.1 or above 10.0 mIU/l average triiodothyronine and thyroxine levels in patients with subclinical thyrotoxicosis were 89.6 and 73.2%, respectively, higher than in patients with subclinical hypothyroidism. Some relationships were found between levels of thyroid hormones and hemodynamic and electrophysiological parameters of the heart. Differential therapy of attacks of atrial fibrillation was effective in 80 and 62.5% of patients with subclinical thyrotoxicosis or hypothyroidism, respectively. PMID- 12891241 TI - [Effect of enteral feeding on exercise tolerance and clinical picture in patients with NYHA class III-IV chronic heart failure]. AB - AIM: To elucidate efficacy of enteral feeding and the role of small intestine in pathogenesis of cardiac cachexia in patients with heart failure and reduced body mass. MATERIAL: Patients with NYHA class III-IV chronic heart failure and deficient body mass (n=34). Control group comprised 32 patients. METHODS: In intervention group standard therapy was supplemented for 24 weeks with balanced nutritional mixture constituting 25% of daily energy requirements. Efficacy of treatment was assessed by 6 minute walk test, evaluation of lean body mass. Studies of intestinal functional activity and biopsies of small intestinal mucosa with morphometrical analysis of bioptates were also performed. RESULTS: Results of 6-minute walk test improved in intervention and control groups by 46.8 and 10.3%, respectively. Average lean body mass increased in intervention group (by 6.7%) and decreased in controls (by 4.9%, p=0.03). Morphometrical study of bioptates revealed augmentation of relative collagen area up to 35-37% (normal value - 10-15%) and pronounced villus atrophy. PMID- 12891242 TI - [Amiodarone-induced keratopathy]. AB - Amiodarone is an iodinated benzofuran derivate class III antiarrhythmic that is a highly effective agent for the prophylaxis and treatment of various cardiac arrhythmias. Amiodarone induced corneal deposits are the most common reversible side effects (70-100%) in humans. This study was conducted to determine ocular effects of long-term therapy with amiodarone. We studied 298 patients who received oral amiodarone. The patients were 146 males and 152 females (mean age 48,0+/-0,8 years). During follow-up for 1-122 months (mean 27,3+/-1,5 months) signs of keratopathy were found in 280 patients (93,9%). Severity of keratopathy depended on cumulative drug dose and duration of administration. Neither type of deposits impaired visual activity, and their presence was not considered as indication for interruption of amiodarone treatment. Ophthalmological monitoring of long-term oral amiodarone therapy did not reveal other ocular side effects. PMID- 12891243 TI - [Microcirculation in hypertension]. AB - Changes of microcirculatory vascular bed play important role in pathogenesis of essential hypertension. All components of the system of microcirculation including vessels and circulating blood are involved into pathological process. Resistance to blood flow is mostly formed in this system. Resistance depends on length, diameter of microvessels and viscosity of blood. In the process of the disease development the following signs of remodeling at the level of microcirculation emerge: decrease of density of vasculature, modification of the media/lumen ratio, increase of blood viscosity, slowing of angiogenesis, impairment of endothelial function. These phenomena can be both cause and consequence of hypertensive disease. Their heterogeneity depends on a variety of factors (duration, stage of the disease, etc.). It seems feasible to differentiate therapy in accordance with character and severity of changes in the system of microcirculation. PMID- 12891244 TI - [Fibrin glue in cardiovascular surgery]. AB - A variant of fibrin glue has been designed. This glue can be prepared on the basis of cryoprecipitation in departments of blood transfusion (fibrinogen concentration 60-90 g/l). The glue has been used in urgent abdominal and cardiac surgery in particular for facilitation of hemostasis after sternotomy and suturing of the aorta, pulmonary artery and atria. PMID- 12891245 TI - [Results of endovascular revascularization with the use of stents after coronary bypass surgery in patients with ischemic heart disease]. AB - AIM: To compare results of stenting in patients with and without prior coronary artery bypass grafting (CABG). METHODS: In 21 patients after CABG stents were implanted both in venous grafts and native vessels (secondary revascularization) and in 25 patients without history of CABG native vessels were stented (primary revascularization). RESULTS: Primary revascularization: there was 1 periprocedural non Q wave myocardial infarction (4%), angina disappeared in 88% and recurred in 1 year in 24% of patients; one year restenosis rate - 18.2%. Secondary revascularization: 1 periprocedural non Q wave (4.8%) and 1 Q wave (4.8%) myocardial infarction. Angina disappeared after stenting in 90.5% and recurred in 1 year in 19% of patients; one year restenosis rate - 17.6%. CONCLUSION: Both primary and secondary revascularizations were highly effective. However secondary stenting was associated with more frequent acute complications. PMID- 12891246 TI - [Safety of exercise and dobutamine stress echocardiography (results of 2383 tests)]. AB - AIM: To elucidate complications and unfavorable side effects of diagnostic exercise and dobutamine stress echocardiography. MATERIAL: Results of exercise (n=1969) and dobutamine (n=414, 34% with the use of atropine) stress echocardiography conducted in 2 centers for diagnosis of coronary heart disease. Mean age of patients 54,4+/-9,4 years. RESULTS: There were no deaths or life threatening complications. Disturbances of cardiac rhythm and conduction (31% during exercise and 61% during dobutamine stress tests) were most frequent events. One of 13 exercise and of 5 dobutamine stress tests was stopped because of complications or side effects. Of 15 cases of ventricular tachycardia and 16 of hypotension (lowering of blood pressure by 20 mm Hg) 9 and 13, respectively, occurred during dobutamine tests. There were differences between centers in frequency of atropine use during dobutamine tests and therapy of stress induced chest pain. CONCLUSION: The data confirmed that both exercise and dobutamine/atropine stress echocardiography could be safely used in diagnosis of coronary heart disease. PMID- 12891247 TI - [Clinical symptoms of erosive and ulcerous lesions of gastroduodenal zone in patients with unstable ischemic heart disease and the significance of endoscopy in diagnosis]. AB - We studied 103 patients with unstable ischemic heart disease and clinical signs of gastroduodenal injury. Among them 62 (60.2%) patients had moderate gastric dyspepsia and abdominal pain during first 1-5 days of hospitalization. In 17 (16.5%) cases these clinical symptoms were accompanied with symptoms of gastroduodenal hemorrhage. Twenty four patients (23.3%) with history of ulcer disease or chronic gastritis, were asymptomatic. In 96 (93.2%) patients endoscopy performed mainly on days 20-24 of hospital stay detected gastroduodenal erosions and ulcers: acute ulcers were found in 19.4, acute erosions - in 40.8, exacerbations of peptic ulcer - in 33.0% of patients. Clinical symptoms of gastroduodenal ulcers and erosions often did not reflect character, severity and dynamics of pathological process. Therefore gastro- duodenoscopy had decisive importance for diagnosis and assessment of effect of treatment. Endoscopy should be carried out with due consideration of concomitant coronary pathology. PMID- 12891248 TI - [Combination therapy with fluvastatin and fenofibrate in ischemic heart disease patients with combined hyperlipidemia and type 2 diabetes]. AB - AIM: To assess efficacy of combination therapy with fluvastatin and fenofibrate in ischemic heart disease (IHD) patients with combined hyperlipidemia and type 2 diabetes. MATERIAL: Patients with IHD and combined hyperlipidemia with (n=56)) or without type 2 diabetes (n=30). METHODS: After 8-week diet period the patients were randomized to 4 weeks monotherapy with either fluvastatin (40 mg/day) or micronized fenofibrate (200 mg/day). In patients whose low-density lipoprotein cholesterol (LDL CH) remained > 2,6 mmol/1 and triglycerides (TG) > 2.3 mmol/1 combination of fluvastatin 40 mg/day and fenofibrate 200 mg/day was used for the next 12 weeks. RESULTS: Target levels of LDL CH and TG were achieved in 75 and 88%, respectively, of diabetics, and in 73 and 88%, respectively, of non diabetics. CONCLUSION: The use of combination of fluvastatin and fenofibrate was more effective then monotherapy for correction of lipid abnormalities in combined hyperlipidemia both in diabetics and non-diabetics with IHD. PMID- 12891249 TI - [Syndrome of impaired tolerance to dietary fats in coronary heart disease]. AB - Changes of main lipid and apolipoprotein parameters of lipid transport system during postprandial hyperlipidemia were studied in subjects without (n=72) and with (n=167) overt coronary heart disease (CHD) including patients with noninsulin dependent diabetes (n=48). Tolerance to dietary fat was evaluated by oral fat load test (consumption of 639+/-7 ml of 20% cream after fast). In subjects without CHD reaction of lipid transport system to fat load was characterized by lowered total, low density lipoprotein cholesterol, apolipoprotein apo B and apo B/apo A-1 ratio, lowered or unchanged triglycerides, stable high density lipoprotein cholesterol level at the background of apo A-1 elevation by 6-th hour after load. Patients with CHD exhibited signs of impaired tolerance to dietary fat: elevation of triglycerides, apo B and apo B/apo A-1 ratio, lowering of high density lipoprotein cholesterol, increase of percentage of subfractions of small high density lipoproteins particles, increase of triglyceride content in low and high density lipoprotein fraction during 6 hours after fat load. PMID- 12891250 TI - [Selection of protection regimen of dual-chamber cardiac pacing against paroxysmal supraventricular tachycardia]. AB - Criteria of selection of a type of protection against paroxysmal supraventricular tachycardia were studied in 38 patients with permanent dual-chamber cardiac pacing. Three types of protection were distinguished. Criteria for their selection were presence of "pacemaker syndrome", state of chronotropic function of the heart, frequency of attacks of supraventricular tachycardia, results of modeling of pathological atrial activity. During follow-up after programming of optimal type of protection of dual-chamber pacing (mean duration 5.1 years) chronic supraventricular tachycardia developed in 13.5% of patients, survival was 89.2%. PMID- 12891251 TI - [Spatial vector electrocardiography: technique, perspectives of use]. AB - Potentials of the use of computer synthesis of integral electrical vector of the heart D0 are described. Calculation of spatial angular vector velocity and linear velocity of its movement along trajectory can be carried out in a framework of biophysical dipole model. Spatial presentation of vector is realized and its behavior in accordance with established pathologies discussed. Possible diagnostic value of obtained results and utility of their introduction into clinical practice are stressed. PMID- 12891252 TI - [Mobilization of bone marrow stem cells in the management of patients with heart failure. Protocol and first results of ROT FRONT trial]. AB - Mobilization of bone marrow stem cells by granulocyte-colony-stimulating factor (G-CSF) is considered to be an alternative to invasive transplantation of autologous myoblasts or stem cells directly into injured cardiac tissue. We have started a 24 week randomized open study in order to elucidate effects of G-CSF (filgrastim) on clinical, hemodynamic and neurohumoral status of patients with NYHA class II-IV chronic heart failure due to ischemic heart disease with zones of nonviable myocardium and left ventricular ejection fraction <40% as well as to assess safety of addition of G-CSF to standard therapy with ACE inhibitors and beta-blockers. It is planned to include 20 patients into each filgrastim (5 mg/kg/day) and control (0.9% NaCl) groups. Methods to be used: dobutamine stress echocardiography for detection of myocardial viability, magnetic resonance tomography, 6-minute walk test, quality of life questionnaire. By the present time 5 patients were included (4 in filgrastim and 1 in control group) and passed 3-6 months points. A control patient died suddenly on 11th week. All patients in filgrastim group are alive (1 experienced obvious improvement, 2 remained stable, and 1 deteriorated and required urgent hospitalization). None of the patients had signs of appearance of 'regenerated' myocardial zones. The patient with positive clinical dynamics was characterized by young age (48 years), moderately severe heart failure (NYHA class II) and pronounced leukocyte reaction to filgrastim (12 fold increase in white blood cell count with appearance of myelocytes and myeloblasts ). In contrast patients without improvement were older than 60 years, had NYHA class III heart failure and experienced just 6-8 fold increases in leukocyte count. These factors are suggested to be predictors of clinical efficacy of G-CSF in patients with heart failure. PMID- 12891253 TI - [Effect of combined antihypertensive and lipid lowering therapy on the level of coronary risk and tissue insulin resistance in patients with metabolic syndrome]. AB - AIM: To assess effect of combined antihypertensive and lipid lowering therapy on main characteristics of metabolic syndrome and total coronary risk. MATERIAL: Seventy five patients with metabolic syndrome aged 40-59 years were included into an open study with parallel groups in which the following combinations of lipid lowering and antihypertensive drugs were used for sweeksinch: atorvastatin + perindopril (n=17, group 1); simvastatin + atenolol (n=17, group 2); fenofibrate + atenolol (n=21, group 3), and simvastatin + indapamide (n=20, group 4). Systolic and diastolic blood pressures (BP), blood serum concentrations of total cholesterol (CH), low density lipoprotein (LDL) and high density lipoprotein (HDL) CH, glucose and immunoreactive insulin were measured before and after treatment. Total coronary risk was calculated according to the PROCAM model. RESULTS: Treatment was associated with decreases of systolic and diastolic BP (from 11% in group 1 to 18,4% in group 2), lowering of total CH (by one third in groups 1 and 2, by 20% in groups 3 and 4), and triglycerides (by 2 times in group 3 and by 7-21% in other groups). Increases of HDL CH were observed in groups 2-4. There were no significant changes of markers of insulin resistance. Baseline total coronary risk was high (37%). Treatment was accompanied with lowering of total risk by 57.4-70%. CONCLUSION: Combined antihypertensive and lipid lowering therapy leads to pronounced lowering of high coronary risk in metabolic syndrome. PMID- 12891254 TI - [QT interval dispersion and hypertension in different variants of left ventricular geometry (population study)]. AB - AIM: To assess relationship between QT dispersion, hypertension, and different variants of left ventricular geometry (concentric or eccentric hypertrophy, concentric remodeling). MATERIAL AND METHODS: Blood pressure measurement, electro and echocardiography were used in the study of a random sample of Tallinn population (321 men and 396 women) aged 35-59 years. Corrected QT dispersion (QT(c)D) equal to or exceeding 70 ms was considered increased. RESULTS: Increase of QT(c)D was associated with left ventricular hypertrophy, especially with its concentric variant. QT(c)D prolongation was significantly more frequent in subjects with hypertension. PMID- 12891255 TI - [Therapy of affective disorders with tianeptine in patients with essential hypertension]. AB - Tianeptine was given to 30 patients aged 39-54 years with essential hypertension and masked, preclinical (n=29) or clinically evident (n=1) depression. This was associated with improvement of clinical psychiatric parameters and data of standardized questionnaire in 29 patients, and with decrease in frequency of hypertensive crises. PMID- 12891256 TI - [Clinical manifestations and characteristics of vascular bed involvement in atherosclerosis of lower extremities in patients with type 2 diabetes]. AB - Clinical manifestations of arterial insufficiency and characteristics of vascular damage were compared in 68 patients (54 men, 14 women, mean age 64.5+/-1.1 years) with atherosclerosis of arteries of lower extremities including 40 patients with type 2 diabetes. Color duplex scanning was used for assessment of severity of vascular wall involvement. Ankle blood pressure was measured by Doppler technique with calculation of ankle-brachial index. Atherosclerosis of arteries of lower extremities in patients with diabetes manifested clinically as neuropathy while intermittent claudication was more characteristic of patients without diabetes. Severity of medial calcinosis and vascular wall rigidity were related to duration of diabetes. Ankle-brachial index in patients with diabetes was 20-30% and 1.5-2 times higher compared with those without diabetes with similar occlusive and nonocclusive changes of arteries of lower extremities, respectively. Color duplex scanning did not reveal differences between patients with and without diabetes in frequency of large multiple atherosclerotic plaques in abdominal aorta, iliac, popliteal arteries. PMID- 12891257 TI - [Physical exercise and atherosclerosis: dynamic high intensity exercise as a factor inducing exogenous dyslipidemia]. AB - Effect of dynamic exercise of various intensity (maximal 100% and submaximal -- 80, 70 and 60% of individual maximally tolerated work load) on blood lipids, lipo and apolipoproteins (apo) was studied in healthy subjects and patients with coronary heart disease. Both in healthy persons and patients high intensity (100 and 80%) exercise was associated with atherogenic changes of lipid transport system: increases of levels of total and low density lipoprotein cholesterol, triglycerides, apolipoprotein B and apo B/apo-A1 ratio. On the contrary, changes after exercise of moderate (60%) intensity (lowering of apo-B containing lipoproteins in healthy subjects and patients and elevation of apo-A1 concentration in healthy subjects) were antiatherogenic. PMID- 12891258 TI - [Effect of atherogenic risk factors on the state of complex intima-media of common carotid artery]. AB - AIM: To elucidate relationship between presence of atherogenic risk factors (dyslipidemia, hypertension, smoking, etc.) and common carotid artery intima media thickness. MATERIAL AND METHODS: Color duplex carotid artery scanning and determination of blood lipids, lipo- and apolipoproteins were carried out in 160 patients aged 34-73 (mean 57.4+/-8.2) years. RESULTS: Dyslipidemia, hypertension (diastolic blood pressure > or = 95 mm Hg), smoking and intima-media thickness > 1 mm were present in 57, 49, 54 and 65% of patients, respectively. Intima-media thickness increased with age and in the presence of hypercholesterolemia, hypertension and smoking. Significant (p<0.05) correlations existed between intima-media thickness and levels of total cholesterol (r=0.512), triglycerides (r=0.411), apolipoprotein B (r=0.497), high density lipoprotein cholesterol (r= 0.404), apolipoprotein-A1 (r=-0.386), duration of hypertension (r=0.488). CONCLUSION: Ultrasound study of intima-media thickness allowed to assess harmful effect of various atherogenic risk factors (mechanical - hypertension, biochemical - dyslipidemia, toxic - smoking) on the state of arterial wall. PMID- 12891259 TI - [Risk factors of ischemic heart disease, data of exercise tests, 24-hour ECG monitoring and coronary angiography in young women with history of myocardial infarction]. AB - Most frequent risk factors in a group of 40 premenopausal women with history of myocardial infarction were smoking (70%) and hypertension (62.5%). Reaction to bicycle exercise tests was negative in 42% and ischemic (mainly pain combined with ST-segment changes) in 50% of these women. Ischemic ECG changes were found during both exercise test and 24-hour ECG monitoring in 30% and were not found at all in 40% of women. Angiography revealed coronary artery stenoses in 58% of women. PMID- 12891260 TI - [Assessment of coronary reserve in patients after myocardial infarction]. AB - Clinical data were compared with results of stress tests (veloergometry and transesophageal cardiac pacing) in 320 patients 1.5-2 months after myocardial infarction. Severity of angina was classified as functional class 0, I, II, III and IV in 78 (24.4%), 162 (50.6%), 64 (20%), 12 (3.75%), and 4 (1.25%) patients, respectively. Characteristics of coronary reserve by exercise test and transesophageal pacing considerably differed mostly in patients with class 0 and I and were practically identical in patients with class III and IV angina. Pearson's coefficient of correlation was the highest between clinical data and results of transesophageal pacing. It was lower between results of pacing and veloergometry and the lowest between clinical data and results of veloergometry. Values of Pearson's coefficient of correlation rose with increase of functional class of angina from 0.38 to 0.88. All methods analyzed in this study were reliable for estimation of degree of lowering of coronary reserve in patients with high functional class of angina. However in patients without angina or with class I angina additional methods were required and transesophageal cardiac pacing appeared to be a preferable technique for assessment of level of coronary reserve in these patients. PMID- 12891261 TI - [Is there an alternative to traditional stress tests? (One more time about transesophageal cardiac pacing)]. AB - AIM: To assess diagnostic potential of transesophageal cardiac pacing in patients with ischemic heart disease. MATERIAL: One hundred patients with overt or suspected ischemic heart disease (IHD) unable to perform or complete standard exercise test. METHODS: In addition to clinical investigation and transesophageal cardiac pacing stress Doppler echocardiography was carried out in 60 and coronary angiography in 30 patients. RESULTS: Transesophageal pacing was completed in 95% of patients. Sensitivity of the test for diagnosis of IHD was 85%, specificity- 77%, predictive value of positive and negative results--95 and 73%, respectively. Combined use of transesophageal pacing and Doppler echocardiography increased sensitivity of diagnosis of IHD up to 92%. Blood pressure increment during transesophageal pacing did not exceed 10 mm Hg. CONCLUSION: Transesophageal cardiac pacing is a highly informative, simple, safe and widely available selective stress test. Results of this test does not depend on concomitant pathology and physical fitness of a patient. Therefore it presents an alternative to standard stress tests and is a method of choice in many diagnostic situations. PMID- 12891262 TI - [Some unsolved problems of assessment of blood pressure level]. AB - The paper deals with blood pressure level assessment and selection of individual optimal blood pressure by means of determination of critical frequency of fusion of flashes. The problem of "white coat" and office hypertension as well as encoding of these states in the context of WHO International Classification of Diseases is also discussed. PMID- 12891263 TI - [Blocker of angiotensin-1 receptor antagonist losartan. Part III. Experience in therapy for chronic heart failure and after acute myocardial infarction]. AB - This is the third part in a series of papers dealing with various aspects of clinical pharmacology of the first AT(1)-receptor antagonist losartan and its therapeutic use in hypertension, diabetic nephropathy, chronic heart failure, and acute phase of myocardial infarction. This part contains review of literature data concerning the use of losartan for the treatment of chronic heart failure and high risk patients after myocardial infarction. Efficacy and safety of losartan in these conditions was demonstrated in two major randomized trials. In one of these trials effects of the drug were studied in patients with chronic heart failure and in the second one therapy with losartan was started in acute period of myocardial infarction. PMID- 12891264 TI - [Arterial switch operation in reconstructive surgery]. AB - Arterial switch is a relatively novel surgical approach aimed at augmentation of blood flow at the account of less important arterial branches. The method consists in suturing vessels of different diameter end to end and end to side. Arterial switch is used for perfusion of donor organs, kidney in particular. The authors suggested a variant of arterial switch--intrapericardial subclavian to pulmonary arterial anastomosis--in the presence of stenosis of right ventricular outflow tract in tetralogy of Fallot. This anastomosis has several advantages over classical methods of surgical correction of Fallot's tetralogy. Coronary artery bypass surgery using mammary artery grafts is based on arterial switch principle and represents the most effective technique of treatment of coronary artery stenoses. Multiple experimental and clinical methods of arterial switch having been proposed and investigated by the authors since 1955 have found wide application in reconstructive surgery. PMID- 12891265 TI - [Structural and functional changes of the myocardium after coronary artery bypass surgery in patients with postinfarction left ventricular remodeling]. AB - In patients with left ventricular end diastolic dimensions >5--< or =5.5 cm myocardial revascularization was associated with significant decreases of diastolic and systolic left ventricular dimensions in immediate postoperative period. This adaptational phenomenon was called passive reducing left ventricular remodeling. In patients with left ventricular end diastolic dimensions <5 and >5.5 cm revascularization was associated with dilation of the left ventricle. Restoration of myocardial blood supply produced overall positive effect on left ventricular functional capacity in patients with various severity of its postinfarction remodeling. Pathways of realization of this positive effect could be different. PMID- 12891266 TI - [Clinical and hemodynamic characteristics of ischemic heart disease in patients with deficient body mass]. AB - AIM: To elucidate clinical and hemodynamic characteristics of patients with ischemic heart disease and low body mass. MATERIAL: Patients with stable angina (n=162) divided into 3 groups according to body mass index (below 25, 25-27 kg/m2 and above 27 kg/m2). RESULTS: Patients with low body mass index compared with those with intermediate values had worse clinical (arrhythmias, derangements of conduction, recurrent myocardial infarctions, cases with heart failure), hemodynamic (lower stroke volume and cardiac index, higher total peripheral resistance), and cardiometric (lower ejection fraction, larger diastolic volumes, greater relative myocardial mass) characteristics. CONCLUSION: These findings constitute a pathophysiological substrate for increased risk of death in patients with ischemic heart disease and deficient body mass. PMID- 12891267 TI - [Effect of restoration of coronary blood flow on dynamics of myocardial perfusion and function in patients with acute myocardial infarction. Data of gated technetium-99m-sestamibi SPECT]. AB - Effects of coronary blood flow restoration on dynamics of myocardial perfusion and function were studied in 50 patients with acute myocardial infarction during 6-month follow-up. Gated SPECT with 99mTc-MIBI (G-SPECT) was performed before onset of reperfusion procedures, in 24 hours, 10 days, 1 and 6 months. In patients with successful reperfusion (n=36) perfusion defect (PD) size decreased by 6.78+/-0.97% (p<0.001) and its severity by 221.3+/-85 std (p<0.001) in 24 hours. In 10 days PD size decreased by 3.9+/-0.88% (r<0.05) and severity by 149+/ 39 std (p<0.05), whereas in 1 month PD size decreased by 2,18+/-1,4% (p<0.05), PD severity by 146.3+/-67 std (p<0.05). Successful reperfusion was associated with improvement of contractility: left ventricular ejection fraction increased by 3% (p<0.05) on day 10 of the disease. In patients with failed reperfusion PD size and severity decreased in 10 days by 4.8+/-4.2%, p<0.05, and 276+/-75 std, p<0.05, respectively, in 6 months PD size decreased by 3.1+/-0.9%, p<0.05, left ventricular ejection fraction did not change during follow-up. We conclude that successful reperfusion therapy promotes restoration of myocardial perfusion and improvement of its contractility. PMID- 12891268 TI - [Dynamics of parameters of left ventricular diastolic function in elderly patients with myocardial infarction]. AB - Dynamics of parameters of intraventricular filling flows were studied in 83 men (46 older and 37 younger than 60 years) admitted within 12 hours after onset of Q wave myocardial infarction. Echocardiography was carried out on days 1, 3, 5, and 21 of hospitalization with registration of standard characteristics of left ventricular systolic and diastolic function as well as velocity and time of propagation of intraventricular filling flow. At initial study older patients more often had cardiac failure and ventricular dysfunction. During treatment younger patients had more noticeable decrease of active myocardial relaxation and increase of E/FPV reflecting elevation of intraventricular pressure. Left ventricular remodeling (increases of ventricular diameter and sphericity index) and dilatation were more pronounced in older patients. PMID- 12891269 TI - [Efficacy of combined use of fosinopril and propranolol in acute myocardial infarction]. AB - Echocardiography, Doppler echocardiography and ECG mapping were used in 40 patients aged 30-70 years with first anterior Q-wave myocardial infarction (MI) for the study of effects of combination of fosinopril and propranolol (20 patients) in comparison with propranolol alone (20 patients) on hemodynamics and size of myocardial damage. The use of fosinopril and propranolol was associated with lowering of systolic and diastolic blood pressure without development of hypotension, reduction of sum, number of leads with, and amplitude of ST-segment elevations, and improvement of parameters of left ventricular function. In propranolol treated patients favorable changes of all these parameters were less pronounced. Inhospital clinical course of the disease was also better in patients treated with fosinopril and propranolol. PMID- 12891270 TI - [Physical exercise and atherosclerosis: proatherogenic effects of high and moderate intensity static exercise on blood lipid transport system]. AB - Effects of high (100 and 80 kg) and moderate (60 kg) intensity static leg exercise on blood serum lipoproteins and apolipoproteins (apo) A1 and B were studied in healthy subjects (n=11) and patients with coronary heart disease and class I angina (n=11). Static leg exercise with loads exceeding 60 kg were associated with atherogenic changes of blood lipid transport system: elevation of levels of triglycerides, apoprotein B and apo B/A ratio both in healthy subjects and patients, and of total and low density lipoprotein cholesterol in patients. These post exercise changes were more pronounced in the presence of fasting hyperlipidemia and their severity increased with increase in duration of exercise. Static leg exercise did not increase concentration of high density lipoprotein cholesterol. For prevention of post exercise atherogenic dyslipidemia it is expedient to supplement strength training programs with dynamic exercise of moderate intensity. PMID- 12891271 TI - [Clinical and metabolic effects of cardioselective beta-adrenoblockers nebivolol and metoprolol in patients with hypertension and ischemic heart disease associated with type 2 diabetes]. AB - AIM: To compare efficacy and safety of cardioselective beta-adrenoblockers nebivolol and metoprolol in patients with hypertension and ischemic heart disease (IHD) combined with type 2 diabetes. MATERIAL AND METHODS: Patients with IHD, class II-III angina and moderately severe compensated or subcompensated type II diabetes (n=35, 29 with grade 1-2 hypertension according to WHO 1999 classification) were divided into 2 groups with similar clinical and laboratory characteristics. Patients of group 1 received metoprolol (50-100 mg/day), of group 2--nebivolol (5-7.5 mg/day) for 8 weeks. Data of physical investigation, exercise tests, 24-hour blood pressure and heart rate monitoring, analysis of lipid spectrum and glycemic profile were used for assessment of treatment efficacy. RESULTS: Decrease of number of anginal attacks and nitroglycerine requirement was more pronounced in nebivolol treated patients. The use of both drugs was associated with lowering of blood pressure and heart rate. There were no unfavorable changes of parameters of carbohydrate and lipid metabolism. Pronounced (by 19.2%) lowering of triglycerides was observed in nebivolol treated patients. Both drugs were well tolerated. CONCLUSION: Nebivolol and metoprolol are effective and safe antianginal agents in patients with IHD and hypertension combined with type 2 diabetes. However in doses used nebivolol produced more favorable metabolic and hemodynamic effects. PMID- 12891272 TI - [Polymorphic markers I/D and G7831A of angiotensin-I-converting enzyme gene and left ventricular hypertrophy in patients with essential hypertension]. AB - AIM: To elucidate possible association of angiotensin-I-converting enzyme (ACE) gene polymorphic markers I/D and G7831A with left ventricular hypertrophy (LVH) in patients with essential hypertension. MATERIAL: Patients with essential hypertension (n=123, 37 with and 86 without LVH, mean age 59.15+/-1.19 years). METHODS: Left ventricular (LV) mass was determined echocardiographically by Devereux method. Alleles and genotypes of ACE gene polymorphic markers were identified by polymerase chain reaction. RESULTS AND CONCLUSION: There was no association between I/D marker of ACE gene and LVH. Carriers of A allele compared with carriers of G allele of G7831A marker had significantly higher LV mass (284.1+/-10.20 g, and 248.5+/-14.42 g, respectively, p=0.033) and LV mass index (151.7+/-5.23 g/m2 and 131.0+/-6.74 g/m2, respectively; p=0.02). Among patients with LVH frequency of A allele was significantly higher than among patients without LVH (0.401 and 0.230, respectively; p=0.0065, OR=2.116 [1.197-3.7481]). Using binary logistic regression model we have found that presence of LVH was linked with age, sex and maximal systolic blood pressure (BP). Such factors as smoking, maximal diastolic BP, ordinary systolic and diastolic BP, duration of hypertension, coronary artery disease and diabetes were not related to LV mass index. Using multifactorial logistic regression model we have found that the presence of A allele of G7831A polymorphic marker of ACE gene, age and maximal systolic BP could be considered as independent risk factors of LVH. PMID- 12891273 TI - [Twenty-four hour blood pressure rhythm in patients with hypertension and ischemic heart disease]. AB - AIM: To elucidate relationship between presence of ischemic heart disease and 24 hour blood pressure (BP) profile. MATERIAL AND METHODS: Twenty four hour BP index characterizing degree of nocturnal blood pressure fall was measured in patients with hypertension without coronary heart disease (n=65, group 1), with hypertension and effort angina (n=35, group 2), with effort angina and normal BP (n=25, group 3). RESULTS: Values of 24-hour BP systolic and diastolic index were in group 1 12.7+/-1.3 and 16.0+/-1.1%, respectively, in group 2 5.8+/-2.4 and 7.4+/-3.6%, respectively, in group 3, 6.0+/-2.2 and 6.0+/-3.0%, respectively (p<0.05, groups 2 and 3 vs group 1). CONCLUSION: Decreased nocturnal BP lowering could be a compensatory reaction aimed at maintenance of adequate blood flow through stenosed coronary vessels. PMID- 12891274 TI - [Effect of nebivolol on the state of pituitary-gonadal system and lipid peroxidation in young and middle aged men with hypertension]. AB - Nebivolol (5-10 mg/day) was given for 12 weeks to 32 patients aged 30-65 years with mild and moderate hypertension. Parameters of pituitary-gonadal system, lipid peroxidation and antioxidant system were determined before nebivolol, and after 1 and 3 months. Normalization of diastolic blood pressure effect was achieved in 87.5% of patients with maximal blood pressure lowering (by 18-20%) after 3 months. Administration of nebivolol was associated with decreases of follicle-stimulating and luteinizing hormones, significant elevation of testosterone concentration (p<0.001), suppression of processes of lipid peroxidation and improvement of activity of antioxidant system. The drug was well tolerated. PMID- 12891275 TI - [Electrocardiographic picture in patients with congenital defect of atrial septum operated upon in adult age]. AB - Electrocardiograms from 30 patients with congenital atrial septal defect operated upon in adult age were analyzed. One of frequent variants of QRS configuration in standard leads was SIqIII type. It was observed in 50% of patients. Most frequent (in 67% of patients) variants of QRS configuration in lead V1 were rSr' and rSR'. Signs of right ventricular hypertrophy were most often found in a group of patients with high pulmonary hypertension. Changes of terminal portion of ventricular complex such as negative or biphasic T waves were found in 29 of 30 patients (in 63% of cases these changes were localized in leads V1-V3). PMID- 12891276 TI - [Anti-arrhythmic therapy: diagnostic possibilities of signal-averaged electrocardiography and heart rate variability]. AB - Effects of monotherapy with class IC, II and III antiarrhythmic drugs on parameters of signal averaged (SA) ECG and heart rate variability were studied in 88 patients (mean age 45.6+/-7.8 years). Class IC drugs (ethacizine, disopyramide) caused worsening of qualitative parameters of SA ECG and appearance of ventricular late potentials. Therapy with beta-adrenoblockers, amiodarone and sotalol in patients with ventricular arrhythmias was associated with improvement of parameters of SA ECG, lowering of sympathetic and augmentation of parasympathetic activity without sings of arrhrythmogenic and negative inotropic effects. Combination of noninvasive diagnostic methods including SA ECG, temporal and spectral analysis of heart rate variability, Holter ECG monitoring can facilitate selection of appropriate antiarrhythmic therapy and control of its efficacy. PMID- 12891277 TI - [Stent thrombosis in patients with myocardial infarction]. AB - The problem of stent thrombosis in patients with myocardial infarction is reviewed with consideration of possible mechanisms and causes of in-stent thrombus formation during first 30 days after the procedure. Roles of the patient factor, factor of the stented vessel, stent in self, stent deployment technique, and management after stenting are discussed. PMID- 12891278 TI - [Angiotensin I receptor antagonist losartan. Part II. Effects in arterial hypertension and diabetic nephropathy]. AB - This is the second part in a series of papers dealing with various aspects of clinical pharmacology of the first AT1-receptor antagonist losartan and its therapeutic use in hypertension, diabetic nephropathy, chronic heart failure, and acute phase of myocardial infarction. This part contains review of literature data concerning the use of losartan for the treatment of hypertension and diabetic nephropathy including results of two major randomized trials which for the first time demonstrated ability of losartan to improve long term prognosis in patients with hypertension and diabetic nephropathy. PMID- 12891279 TI - [Romano-Ward syndrome initially diagnosed as epilepsy]. AB - A report of a case of Romano-Ward syndrome presenting as epilepsy is accompanied with discussion of symptoms characteristic of the syndrome and possible causes of erroneous primary diagnosis. PMID- 12891280 TI - [Concept of critical stenosis in cardial surgery]. AB - When stenosis of a large artery reaches about 75% of its lumen blood flow distal to obstruction abruptly falls and ceases to play decisive role for the region it supplies. This region survives due to opening of collaterals. This allows to cross-clamp coronary artery for 20-30 minutes and to create distal anastomosis during aorto-coronary venous or mammary coronary artery bypass grafting without artificial circulation. Another example is mitral stenosis. There exists a critical value of an orifice area (1 cm2 or 25% of the lumen) when abrupt decompensation develops. Similar cut off values of inflow (nourishing) vessel stenoses can be demonstrated in other regions of cardiovascular system. Thus conception of a 'critical stenosis' is universal for the arterial part of circulation and finds wide application in clinical practice. PMID- 12891281 TI - [Markers of inflammation and platelet aggregation in patients with non ST elevation acute coronary syndrome treated with atorvastatin or pravastatin]. AB - AIM: To find out whether early use of atorvastatin and pravastatin in patients with non-ST elevation acute coronary syndrome is associated with rapid changes of platelet aggregation and plasma levels of markers of inflammation. MATERIAL AND METHODS: Ninety patients (<24h from pain onset, age 64+/-10 years) treated with aspirin and heparin were randomized to open atorvastatin 10 mg/day (n=30), atorvastatin 40 mg/day (n=29) or pravastatin 40 mg/day (n=31). Spontaneous and ADP induced platelet aggregation (light transmission), plasma levels of interleukin 6 (IL-6) and C-reactive protein (CRP) (immunoassay) were assessed at baseline, on days 7 and 14. RESULTS: Baseline clinical characteristics, platelet aggregation parameters, CRP and IL-6 levels were similar in all groups. In all groups levels of total and low-density lipoprotein (LDL) cholesterol (CH) were lowered by days 7 (p<0.01) and 14 (p<0.01 vs. baseline and for both atorvastatin groups vs. day 7). Spontaneous platelet aggregation decreased by 15% from baseline, p<0.01, on day 14 in patients receiving atorvastatin 40 and was unchanged in other groups. Changes of ADP induced platelet aggregation, IL-6 and CRP levels were not significant in all groups. However combination of 2 atorvastatin groups (n=59) revealed decrease of CRP by 18% from baseline on day 14 (from 6.94+/-0.97 to 4.76+/-0.76 mg/l, p=0.028). No correlations were found between changes of LDL CH and those of other parameters. CONCLUSION: In otherwise conventionally treated patients with non-ST elevation acute coronary syndrome early use of atorvastatin was associated with rapid (in 14 days) decrease of CRP level. Higher dose of atorvastatin (40 mg/day) induced favorable changes of spontaneous platelet aggregation. There were no significant changes of parameters studied in pravastatin treated patients. PMID- 12891282 TI - [Structural-geometric changes of the myocardium and the state of central hemodynamics during left ventricular remodeling after myocardial infarction]. AB - Intergroup analysis of parameters of central hemodynamics in the process of increase of left ventricular dimensions has shown that changes of pump function of the heart correspond to stages of left ventricular remodeling after myocardial infarction. Decreases of ejection fraction and cardiac output occur in patients with postinfarction aneurysm. At various stages of pathogenesis of remodeling multiple factors facilitate adaptation to novel conditions of functioning. They include structural reorganization of ventricular wall, hypertrophy of myocytes, and augmentation of muscular mass without wall thickening (exocentric hypertrophy). At other equal conditions characteristic for the patients studied starting point of maladaptational left ventricular remodeling and consequent development of cardiac failure can be value of left ventricular diastolic volume 182.7+/-3.28 ml. This volume according to data obtained by the authors represents a threshold over which 'rescue' for the body Staring law is not able to maintain adequate stroke volume and cardiac output. PMID- 12891283 TI - [Clinical effectiveness and safety of citalopram in patients with depression after myocardial infarction]. AB - Citalopram (20 mg/day for 42 days) was given to 20 patients (mean age 54.4+/-2.5 years) with depression after myocardial infarction. This was associated with substantial antidepressive effect (50% and more lowering of the total score of the Beck depression questionnaire) in 89% of patients, reduction of number and severity of somatic complaints, and improvement of parameters of quality of life. Citalopram did not affect blood pressure and according to Holter ECG monitoring data produced no arrhythmogenic or proischemic effects. Overall tolerability of citalopram was good however 21% of patients experienced slight drowsiness, dizziness or sweating. PMID- 12891284 TI - [Relationship between changes of endothelial vasodilator function, vascular and cardiac structure and derangements of blood pressure 24-hour profile]. AB - AIM: To assess relationship between 24-hour blood pressure (BP) rhythm and structural and functional state of the heart and blood vessels. MATERIAL AND METHODS: Echocardiography, sonography of carotid artery intima-media, and assessment of endothelium dependent (flow mediated) and endothelium independent (nitroglycerine) brachial artery vasodilatation were used in the study of 83 patients with stage I-II essential hypertension (mean age 49.2+/-6.2 years). Seventy four patients received indapamide and enalapril for 24 weeks. RESULTS: According to results of 24-hour BP monitoring the patients were divided into 2 groups with normal (dippers) and abnormal (non-dippers) BP rhythm. Non-dippers compared with dippers had more pronounced lowering of endothelium-dependent autoregulation of arterial tone, bigger left ventricular mass thicker carotid artery wall. Linear correlation was found between changes of 24-hour BP rhythm and structural and functional parameters of the heart and vessels. More pronounced regression of their derangements occurred during antihypertensive therapy in those patients in whom normalization of 24-hour rhythm was achieved. PMID- 12891285 TI - [Blood pressure 24-hour rhythms and variability in adolescents with hypertension]. AB - Twenty-four hour blood pressure (BP) monitoring was used for the study of BP 24 hour rhythms and variability in adolescents with hypertension and their normotensive counterparts. The data obtained evidenced for active participation of sympatho-adrenal system in genesis of juvenile hypertension: hypertension was predominantly systolic, BP elevation as a rule was associated with increased heart rate and pulse pressure. Episodes of BP elevation were registered mostly during day time. Desynchronization of 24-hour rhythms due to pronounced BP lowering during sleep characteristic for this age group found its reflection in values of nocturnal BP dipping. More than 1/3 of adolescents with episodes of elevated BP had 'white coat' hypertension. PMID- 12891286 TI - [Association of a polymorphic marker C825T of the beta(3) subunit of G-protein with myocardial hypertrophy in patients with hypertensive disease]. AB - AIM: To elucidate association of a polymorphic marker C825T of the beta(3) subunit of G-protein with myocardial hypertrophy in hypertension. MATERIAL AND METHODS: Alleles of polymorphic marker C825T of the CNB3 gene were identified by polymerase chain reaction with subsequent cleavage of the product by BseDI restrictase in 135 patients with hypertensive disease (essential hypertension). Left ventricular mass was determined by echocardiography. RESULTS: Patients with and without left ventricular hypertrophy had similar frequencies of T allele of the gene studied. Moreover patients with CC, CT and TT genotypes had similar left ventricular mass index (122.3+/-29.8, 118.8+/-29.9, and 115.2+/-18.3 g/m(2), respectively). CONCLUSION: No association exists between polymorphic marker C825T of the CNB3 gene and left ventricular hypertrophy in patients with essential hypertension. PMID- 12891287 TI - [Medico-ecological monitoring of cardiovascular diseases in the urbanized north]. AB - Aim of the study was to elucidate relationship between complex effect on organism of unfavorable ecological factors of high altitudes and course of ischemic heart disease (IHD) and hypertensive disease in Surgut (Khanty-Mansi Autonomous District). Five year medico-ecological monitoring established direct relation between dynamics of average monthly hospital admissions of patients with IHD and hypertension, level of atmospheric pressure and concentration of phenol in the air. Complex of nature conservation measures resulted in decreases of frequency of hospitalizations as well as cardiovascular morbidity with temporary loss of working capacity. PMID- 12891288 TI - [Effects of verapamil on atrial fibrillation spontaneous initiation in the intact canine heart]. AB - L-type Ca(2+) current (I(Ca,L)) has been shown to play a crucial role in initiation of early after depolarization (EAD) in cardiomyocytes. To study the possible role of EAD in spontaneous initiation of atrial fibrillation (AF), we tested the effects of L-type Ca(2+) channel blocker verapamil in canine models of cholinergic-dependent AF. In anesthetized open-chested dogs (n=13) spontaneous AF was induced by two methods: (1) perfusion with acetylcholine (ACh) in normal Tyrode solution at 9 ml/min into the sinus node artery (SNA) and (2) tonic stimulation of the right cervical vagus nerve (5 sec train). In the control, AF was induced in all dogs by perfusion with ACh (2.9+/-0.8 microM, mean+/-SEM) in 96+/-4% of attempts and by vagal stimulation (VS, 59+/-8 Hz) in 74+/-9% of attempts. Verapamil (0.2 mg/kg i.v.) did not alter the AF inducibility both during ACh perfusion and during VS (93+/-4% and 77+/-13%, NS, respectively) in dogs that retained sinus rhythm (n=8). However, verapamil significantly decreased AF inducibility to 50+/-4% and 21+/-13%, respectively, in dogs that passed to AV rhythm (n=5). Verapamil increased duration of both ACh- and vagally-mediated AF from 15+/-2 sec and 15+/-2 sec to 34+/-6 sec and 23+/-4 see (p<0.05 vs. control), respectively. The activation mapping (112 unipolar electrodes) during the initiation of AF did not reveal a difference in epicardial activation patterns before and after verapamil treatment. Inhibiting the I(Ca,L) by verapamil resulted in significant (p<0.05 vs. control) decrease in systolic and diastolic blood pressure, PQ interval prolongation and slowing down of the sinus rate. Verapamil did not affect atrial effective refractory period (AERP) and conduction velocity in the right atria. The reduction of AERP and the deceleration of heart rate by VS (8 Hz) remained unchangeable after verapamil treatment in comparison to control. Thus, the data suggest that the mechanism of spontaneous AF initiation during increased cholinergic activity is not related to EAD in atria. PMID- 12891289 TI - [Effects of metabolic substrates and mannitol on efficiency of cardioplegic protection in isolated rat heart]. AB - The aim of this work was to study rationality of addition of aspartic acid, phosphocreatine, mannitol and tris(bydroxymethyl) aminomethane (trisamine) to a sanguineous cardioplegic solution. Isolated perfused rat hearts were subjected to 40-min normothermic total ischemia and 30-min reperfusion. Cardioplegic solutions were infused for 5 min prior to ischemia. A modified Ringer solution with 25 mM KCI was used as control. Osmolarity and pH of cardioplegic solutions were 340+/-5 mOms and 7.6+/-0.1 at 22 degreesC, respectively. Efficiency of myocardial protection was evaluated by recovery of contractile and pump function during reperfusion. The optimal solution contained aspartic acid (21.5 mM), mannitol (20.0 mM) and trisamine (5 mM). By the end of reperfusion the heart protected by this solution showed almost complete recovery of coronary flow (98+/-3% of the initial value vs. 77+/-3% in the control), and 2.6-fold higher recovery of stroke volume compared to the control. As a result, recovery of external cardiac work index, calculated as cardiac output-mean perfusion pressure, was 64+/-1% of the initial value vs. 24+/-5% in the control. Increase in buffer capacity of this cardioplegic solution by trisamine (up to 20.0 mM) as well as addition of phosphocreatine (10.0 mM) did not result in further augmentation of cardiac function recovery. The results suggest promising perspectives for development of medicinal form of this solution. PMID- 12891290 TI - [Hereditary factors and left ventricular hypertrophy]. AB - Left ventricular hypertrophy (LVH) is an independent risk factor for morbidity and mortality from cardiovascular disease in men and women with hypertension and in asymptomatic subjects with normal blood pressure. In hypertensive patients it is a stronger coronary risk factor than casual blood pressure readings. Correlation between levels of high blood pressure, duration of hypertension and left ventricular mass is poor. Epidemiological studies suggest that left ventricular hypertrophy may be influenced by genetic factors. In our review we present study groups of genes contributing to the development of left ventricular hypertrophy: 1) genes that encode components of hormonal pathways, 2) genes of key sympathetic and parasympathetic receptors, 3) genes that modify intracellular ion homeostasis, 4) genes that modify energy metabolism, 5) genes that modify motor unit composition and regulation. Angiotensinogen gene, angiotensin converting enzyme gene, angiotensin receptor type 1 gene, aldosterone synthase gene, nitric oxide synthase gene, type A natriuretic peptide receptor gene, beta(2)-adrenergic receptor gene, G-protein beta(3) subunit gene are associated with left ventricular hypertrophy. PMID- 12891291 TI - [Angiotensin1-receptor antagonist losartan. Part I. Basic clinical pharmacology]. AB - This is the initial part in a series of papers dealing with various aspects of clinical pharmacology of the first AT(1)-receptor antagonist losartan and its therapeutic use in hypertension, diabetic nephropathy, chronic heart failure, and acute phase of myocardial infarction. Mechanisms of action, pharmacological effects of losartan and other AT(1)-receptor antagonists, as well as data on pharmacokinetics of losartan, its long term tolerability and safety are reviewed in this communication. PMID- 12891292 TI - [Possibilities of stress-echocardiography in diagnostics of pathology of mitral valve. Part II]. AB - The paper deals with different aspects of application of stress-echocardiography for assessment of mitral valve disease (acquired stenosis or insufficiency, mitral regurgitation due to ischemic heart disease and mitral valve prolapse). Continuity equation is preferable to pressure half-time and resistance for estimation of hemodynamic significance of mitral stenosis. In some patients studies at rest reveal only slight mitral regurgitation while during stress it becomes hemodynamically significant and associated with pronounced elevation of pulmonary artery pressure. Deterioration of left ventricular function in this pathology often occurs before appearance of symptoms and even before increase of left ventricular dimensions and lowering of ejection fraction can be registered by echocardiography at rest. In patients with mitral valve prolapse stress induced mitral regurgitation has high predictive power for unfavorable clinical course. It is necessary to remember that in this group of patients abnormalities of left ventricular function during stress can be found in the absence of ischemic heart disease or pronounced regurgitation. PMID- 12891293 TI - [Non ST elevation acute coronary syndrome. Parameters of fibrinolysis during short term use of ticlopidine or clopidogrel]. AB - Whether thienopyridines (ticlopidine or clopidogrel) produce similar effects on fibrinolysis in patients with non ST elevation acute coronary syndrome (NSTEACS) was not well elucidated. AIM: To study changes of parameters of fibrinolysis during short term use of ticlopidine and clopidogrel in NSTEACS patients treated with aspirin and antithrombin. MATERIAL: Patients with NSTEACS treated with aspirin and unfractionated heparin (UFH, n=37) or enoxaparin (n=19). METHODS: The UFH treated patients were randomized to ticlopidine (1000 mg/day for 2 and then 500 mg/day for 5 days, n=19) or no ticlopidine (n=18). Enoxaparin treated patients were randomized to either clopidogrel (300 mg/day for 1 and then 75 mg/day for 6 days, n=10) or no clopidogrel (n=9). Levels of tissue plasminogen activator (TPA) antigen, D-dimer, and activity of plasminogen activator inhibitor (PAI) were measured before and in 24 hours, 3, 7, 14 days after randomization. RESULTS: At baseline values of parameters studied were similar in each pair (thienopiridine - control) of patients groups. Compared with their controls ticlopidine treated patients on day 7 had less pronounced lowering of PAI activity (13.6 and 8.2 U/l, p<0.05) and lower D-dimer concentration (515 and 770 ng/ml, respectively, p<0.05). Clopidogrel treated patients on days 3, 7 and 14 had higher levels of TPA both compared with controls (25.7 and 20.2, p<0.05; 26.5 and 12.9, p<0.01; 24.6 and 15.7 ng/ml, p<0.01; respectively) and baseline. D dimer levels in these patients on same time points were also higher than in controls (969 and 702, p<0.05, 970 and 575, p<0.01, 806 and 484 ng/ml, p<0.01, respectively). CONCLUSION: Compared with controls the use of ticlopidine in patients with NSTEACS treated with aspirin and UFH was associated with less pronounced lowering of PAI activity and lower level of D-dimer. This could be interpreted as consequence of inhibition of fibrinolysis. The use of clopidogrel in similar patients treated with aspirin and enoxaparin was associated with elevated levels of TPA and D-dimer what presumably reflected augmentation of fibrinolytic activity. PMID- 12891294 TI - [Autonomic tests in survivors of acute phase of myocardial infarction. Feasibility, tolerance, and safety]. AB - In order to assess practicability and safety of simple autonomic tests in survivors of acute phase of myocardial infarction (MI) attempts to carry out active orthostatic test, tests with controlled breathing (6 and 15 breaths per min), and Valsalva maneuver were performed in 210 patients on days 4-11 of MI. All patients had no signs of severe heart failure, were in sinus rhythm and did not interrupt standard therapy which included beta blockers. Tests were not completed in 14 patients (4.8%): orthostatic test in 6 due to hypotension, Valsalva maneuver in 3 because they did not reach required pressure in respiratory airways, and controlled breathing in 6 because of shortness of breath and substernal pain. All symptoms disappeared spontaneously and none of them was accompanied by ECG changes. All other tests were successfully completed without complications. Thus simple autonomic tests used in this study can be safely carried out in most stable patients on days 4-11 of MI. PMID- 12891295 TI - [Assessment of antiischemic efficacy of balloon angioplasty, coronary bypass surgery and conservative therapy after myocardial infarction in patients with single coronary artery disease. Three year prospective study]. AB - Efficacy of balloon angioplasty, coronary bypass surgery and conservative therapy in postmyocardial infarction patients with single coronary artery disease was assessed in 3 year prospective study. Positive clinical effect persisted after coronary artery bypass grafting (CABG) in 78 and 73% of patients and after balloon angioplasty in 60 and 40% of patients in 1 and 3 years, respectively. Significant decreases of number of anginal attacks (by 94.1 and 74.1%, respectively), nitroglycerin consumption (by 93.6 and 74.8%, respectively) and dimensions of ischemic perfusion defects (by 68.8 and 33.7%, respectively) were observed 1 year after both CABG and angioplasty. Lowering of exercise tolerance (by 11.1%), increase of nitroglycerin consumption (by 15.8%) and worsening of myocardial perfusion (by 9%) were observed after 3 years in conservatively treated patients. PMID- 12891296 TI - [Effect of calcium antagonists verapamil and amlodipine on the risk of development of atrial fibrillation after coronary artery bypass grafting]. AB - AIM: To assess effect of calcium antagonists amlodipine and verapamil on the risk of development of atrial fibrillation after coronary artery bypass surgery. MATERIAL: Of 74 patients subjected to mammary artery and venous coronary bypass grafting with the use of cardiopulmonary bypass 19 received amlodipine and 21 - verapamil. RESULTS AND CONCLUSION: Attacks of atrial fibrillation during 24 hour ECG monitoring were registered in 22.9% of patients mostly on days 2 and 3 after surgery. In verapamil treated patients atrial fibrillation occurred 1.5-2 times more often than in amlodipine treated patients or in patients receiving no calcium antagonists. This could possibly be explained by pronounced slowing of atrio-ventricular conduction by verapamil at the background of postoperative general "irritation" of the atrial myocardium. PMID- 12891297 TI - [Surgical treatment of acquired defects and ruptures of interventricular septum]. AB - Surgical repair of posttraumatic ventricular septal defects, aortic-right ventricular fistulas and postinfarction ruptures of interventricular septum was carried out in 15 patients aged 18-75 years. Methods of radical correction included suturing, plastic repair with synthetic and autopericardial patches, and amputation of cardiac apex. Procedures were performed 14 days - 12 months after myocardial infarction and 1-12 months after penetrating knife wounds of the chest and the heart. Duration of follow-up was from 1 month to 9.5 years. There were no cases of defect recanalization and deaths. PMID- 12891298 TI - [Out-of-hospital sudden coronary death: separate acute coronary syndrome or the mode of death due to ischemic (coronary) heart disease?]. AB - BACKGROUND: The existing data on the morphological substrate of out-of-hospital sudden death due to ischemic (coronary) heart disease (IHD) and its relationship to other acute coronary syndromes are not sufficient and even controversial. MATERIAL AND METHODS: We analyzed clinical and pathological data from 170 men who died suddenly (within 6 h) of IHD out of hospital, as well as 54 men who died in hospital a documented acute myocardial infarction (AMI). RESULTS AND CONCLUSIONS: Majority (92,9%) of out-of-hospital sudden coronary deaths were due to AMI and 7,1%t - due to disseminated myocardial micronecroses. Death occurred mostly at early stages of AMI, and predominantly in the presence of morphological signs of acute left ventricular failure (alveolar pulmonary edema). Ruptures of atherosclerotic plaques or endothelial erosions with the intraplaque and superimposed luminal thrombosis were found in 88,8% of AMI cases. In 73,6% of cases plaque ruptures or erosions were located in 75% or greater stenoses mostly in proximal segments of coronary arteries. Acute coronary artery pathology could be identified by examination of serial histological sections, while early irreversible ischemic myocardial injury (early infarction) was detected by light and polarized light microscopy of histotopogrammes of the whole myocardium with due consideration of the complex of signs of cardiomyocyte alteration as well as early inflammatory reaction. PMID- 12891299 TI - [Antiischemic efficacy of trimetazidine in patients with intermittent claudication and effort angina]. AB - Male patients (n=42) with atherosclerosis obliterans and stage IIA-III ischemia of lower extremities combined with class II-III angina pectoris were divided into 2 groups. Patients of group 1 (n=21) were given simvastatin (10-20 mg/day), clopidogrel (75 mg/day) and trimetazidine (60 mg/day) for 6 months, those of group 2 - simvastatin and clopidogrel also for 6 months. Statistically significant decreases of frequency of attacks of angina, nitroglycerine consumption and increases of walking distance occurred only in group 1. PMID- 12891300 TI - [Shortening of PR interval, bradycardia and polymorphic ventricular tachycardia- clinico-electrocardiografical syndrome with high risk of sudden death in children]. AB - AIM: To elucidate clinical and electrocardiographical characteristics of children and adolescents with malignant idiopathic polymorphic ventricular tachycardia. MATERIAL: Patients aged 6-14 years (n=16) with polymorphic bidirectional tachycardia registered either on standard 12-lead ECG (n=6), or during treadmill exercise test (n=2) and Holter ECG monitoring (n=8). None of the patients had overt heart or coronary artery disease, chronic extracardiac pathology or electrolyte disturbances. RESULTS: There was no QT, QT(s) prolongations, or ST segment elevations in right precordial leads. Eleven patients (68.8%) had shortening of PR interval -110 ms without signs of Wolf-Parkinson-White syndrome. All patients with shortened PR had history of syncopal states (9.1+/-9.2 per year), 4 patients (36.4%) had family history of sudden death in young age. Among patients without PR shortening 2 (40%) had history of syncopal attacks (1-2 or 0.6+/-0.89 per year). All patients with short PR had bradycardia while in sinus rhythm (55.5+/-9.1 bpm compared with 78.1+/-2.8 bpm in patients without PR shortening). CONCLUSION: Shortened PR and bradycardia were found to be associated with frequent attacks of syncope in this series of children with polymorphic tachycardia. Therefore combination of short PR, bradycardia and polymorphic ventricular tachycardia was suggested to comprise clinico-electrocardiographic syndrome with high risk of malignant ventricular tachyarrhythmia and sudden death. PMID- 12891301 TI - [Humoral markers of endothelial dysfunction in essential hypertension]. AB - AIM: To elucidate potential markers of endothelial dysfunction and relationships between humoral and vasoregulatory responses of the endothelium in patients with essential hypertension. MATERIAL: Thirty four male patients (mean age 54.4+/-9.2) with I-II degree hypertension and left ventricular hypertrophy. METHODS: Endothelial function was assessed non-invasively by registration of flow-mediated brachial artery dilatation. Serum level of vascular adhesive molecule 1S (VCAM 1S) was measured by enzyme immunoassay, levels of aldosterone and insulin-like growth factor-1 (IGF-1) - by radioimmunoassay. RESULTS: Brachial artery flow mediated dilatation was 2.0+/-7.46%. Patients with hypertension compared with healthy subjects had significantly higher mean levels of VCAM-1S (713.9+/-123.2 and 590.8+/-51.8 pg/ml, respectively, p<0.001), aldosterone (105.41+/-50.17 and 42.7+/-13.16 ng/ml, respectively, p<0.001), and IGF (209.7+/-49.1 and 137+/-21 pg/ml, respectively, p<0.001). Direct significant correlations were found between humoral and functional markers of endothelial dysfunction in the group of patients with essential hypertension. CONCLUSION: These results can be interpreted as indicative of pathogenetic role of IGF-1 and aldosterone in the development of the endothelial dysfunction and myocardial remodeling in essential hypertension PMID- 12891302 TI - [Characteristics of hemodynamics and metabolism and structural polymorphisms of angiotensin converting enzyme and angiotensin II type I receptors genes in women of childbearing]. AB - Parameters of systemic and intracardiac hemodynamics, characteristics of metabolism, and structural polymorphisms of angiotensin converting enzyme (ACE) and angiotensin II type 1 receptor (ATR(1)) genes were studied in women of childbearing age and men of the same age both with first degree hypertension. Women had lower mean 24-hour blood pressure (BP), greater BP variability, preponderance of "dipper" variant of 24-hour systolic and diastolic BP profile, higher prevalence of left ventricular hypertrophy, lower blood levels of cholesterol, triglycerides, urea and potassium. Among women A allele of ATR(1) gene was significantly more frequent than C allele. Comparison of ATR(1) gene polymorphisms in women and men showed that frequencies of AA genotype and A allele were higher in women. Presence of DD genotype of ACE gene in women was related to parameters of both hemodynamics and metabolism, whereas that of II genotype of ACE gene as well as AA and AC genotypes of ATR(1) gene were linked with parameters of hemodynamics. PMID- 12891303 TI - [PRKAR1A gene mutations in two patients with myxoma syndrome (Carney complex)]. AB - Carney complex is an autosomic dominant disorder initially described as the association of cardiac myxomas, spotty skin pigmentation and endocrine overactivity and considered as a multiple neoplasia and lentiginosis syndrome. Mutations in the tumor suppressor gene PRKAR1A, coding for the type 1-alpha regulatory subunit of cAMP-depended protein kinase A have been previously identified in about half of the Carney complex kindreds. In this paper we report identification of the molecular defect in PRKARIA gene in two Carney complex patients. A new mutation (403delAC) located in a 3rd exon of PRKARIA gene has been observed in one case, and a previously described mutation in exon 7 (847delTC) in the second case. PMID- 12891304 TI - [Lyme disease with cardiac involvement]. AB - A case of Lyme disease with cardiac involvement in a patient with congenital kidney pathology is presented. Carditis is one of manifestations of Lyme borreliosis. That is why when myocarditis of obscure etiology is detected in an area endemic for tick borreliosis special laboratory investigation for exclusion of Lyme disease should be carried out. PMID- 12891305 TI - [Non ST elevation acute coronary syndrome: early use of pravastatin or atorvastatin is associated with divergent changes of parameters of hemostasis]. AB - PURPOSE: To find out whether pravastatin and atorvastatin rapidly (in 1-2 weeks) and similarly affect hemostasis in patients with non-ST elevation acute coronary syndrome (NSTEACS). METHODS: Ninety aspirin and heparin treated patients with NSTEACS were randomized <24 hours from pain onset to open pravastatin 40 mg/day (n=31), atorvastatin 10 mg/day (n=30) or atorvastatin 40 mg/day (n=29). At baseline, on days 7, 14 plasma thrombin-antithrombin complex (TAT), prothrombin fragments 1+2 (F1+2), D-dimer, von Willebrand factor (vWF) were measured by ELISA. Results were compared with data from controls (n=18) of another randomized study on similarly treated patients. RESULTS: In all treatment groups levels of low-density lipoprotein cholesterol (LDLCH) were lowered by days 7 (p<0,01) and 14 (p<0,01 vs. baseline and for both atorvastatin groups vs. day 7). In pravastatin group levels of TAT and F1+2 decreased, while vWF level increased. In atorvastatin groups levels of TAT and F 1+2 increased while level of vWF decreased. Contrary to pravastatin group changes in atorvastatin treated patients more resembled those in controls not receiving lipid lowering drugs. Changes of both LDLCH and TCH directly correlated only with changes of vWF (r=0.23, p=0.03 and r=0.25, p=0.02, respectively). No consistent changes of D-dimer occurred. CONCLUSION: Early use of atorvastatin and pravastatin in patients with NSTEACS was associated with rapid divergent changes of some hemostatic parameters. Except lowering of von Willebrand factor changes in atorvastatin treated patients more resembled those in controls not receiving lipid lowering drugs. Von Willebrand factor was the only parameter which changes weakly but significantly correlated with changes of CH and LDL CH levels. PMID- 12891306 TI - [Effect of dipyridamole on blood levels of fibroblast growth factor and vascular endothelial growth factor in patients with ischemic heart disease]. AB - Addition if dipyridamole to standard therapy in patients with ischemic heart disease was associated with improvement of functional class of angina and elevation of blood levels of fibroblast growth factor and Vascular Endothelial Growth Factor. PMID- 12891307 TI - [Prognostication of hemodynamic efficacy of surgical correction of chronic coronary insufficiency based on stress test with nitroglycerine]. AB - In our previous studies we demonstrated possibility of the use of stress echocardiography with sublingual administration of nitroglycerine for detection of viable myocardium. Basing on data about myocardial and cerebrovascular reserves obtained during nitroglycerine tests we created mathematical model for prediction of hemodynamic effectiveness of coronary bypass surgery. Stress echocardiography, Doppler echography of middle cerebral and internal carotid arteries with nitroglycerine were carried out in 80 patients with coronary artery disease before myocardial revascularization. Quantitative equivalent characterizing degree of myocardial reserve preservation in early postoperative period was used as basis for discrimination of patients into groups with positive and negative result of surgery, and coefficients and constants of discriminant function were calculated for these groups. According to results of clinical testing sensitivity, specificity and diagnostic effectiveness of the model were 76.9, 75 and 76%, respectively. PMID- 12891308 TI - [Russian study of efficacy and safety of an angiotensin converting enzyme inhibitor quinapril in patients with hypertension]. AB - Efficacy and safety of an angiotensin converting enzyme inhibitor quinapril were assessed in a 12 week study on 258 patients with 1-2 degree hypertension and various concomitant pathology. The use of quinapril as monotherapy (10-40 mg/day) or in combination with hydrochlorothiazide (12.5 mg/day) was associated with blood pressure (BP) normalization (<140/90 mm Hg) or more then 10% BP lowering in 88% of patients. In the end of the study average blood pressure was 134/84 mm HG. Quinapril was well tolerated. These results allow to recommend the scheme of quinapril use utilized in the study for wide application for the treatment of outpatients with hypertension. PMID- 12891309 TI - [Isradipine-SRO: antihypertensive efficacy, central and peripheral hemodynamic effects in patients with hypertensive disease]. AB - AIM: To assess antihypertensive efficacy of isradipine, to study its effects on central and peripheral hemodynamics, and to investigate relationships between state of microcirculation and response to treatment. MATERIAL AND METHODS: Isradipine was given for 12 weeks to 30 patients (16 men, 14 women, mean age 50.2+/-9.8 years) with I-II stage mild and moderate hypertension. Twenty four hour blood pressure monitoring, assessment of parameters of central hemodynamics and state of microcirculation were carried out before and after 12 weeks of isradipine administration. RESULTS: Administration of isradipine was associated with improved sense of well-being and significant lowering of 24-hour, diurnal and nocturnal systolic/diastolic blood pressure (by 6.4/12.7, 7.6/9.4, 10.5/13.4%, respectively). Average 24-hour heart rate increased mainly during daytime (by 10.6%). Mean hemodynamic blood pressure was lowered by 16.3% due to reduction of total peripheral vascular resistance. There occurred redistribution of types of microcirculation with increase of proportion of normocirculatory and decrease - of spastic types. Changes of parameters of microcirculation at rest and during functional tests evidenced for resolution of baseline arteriolar spasm, stasis and venous congestion in microcirculatory vascular bed. Patients who responded to isradipine more often had spastic (50%) and congestive (35%) types of microcirculation. Patients with hyperemic type did not respond to isradipine. Most frequent side effect was flushing (23.3%) because of which 2 patients stopped taking the drug. PMID- 12891310 TI - [Ramipril in elderly patients with mild and moderate hypertension. Clinical efficacy, effect on cerebral blood flow and intellectual functioning]. AB - AIM: to study different ramipril dosage regimens on cerebral blood flow and cognitive function in elderly patients with mild to moderate hypertension. MATERIAL: 30 out-patient hypertensive patients (13 men, 17 women, age 55-75 years, mean 62+1,5). METHODS: After withdrawal of previous antihypertensive therapy patients were randomized to receive gradual (group 1) and more active (group 2) ramipril regimen. The starting dose (2,5 mg in group 1 and 5 mg in group 2) was increased at 2-week intervals to 10 mg daily. If target blood pressure (BP) level was not achieved hydrochlorothiazide (25 mg/day) was added. The treatment duration was 8 weeks. 24-hour BP monitoring, duplex scanning of brachiocephalic arteries were performed at baseline, after the first dose of ramipril, and after 8 weeks of therapy. Intellectual functioning was evaluated with 4 tests of memory and attention assessment. RESULTS AND CONCLUSIONS: Both single dose and long term ramipril treatment provided stable and steady hypertensive effect throughout 24 hours. There was no case of first dose hypotension in elderly patients. Both starting doses (2,5 mg and 5 mg) were similarly safe. Neither first dose of ramipril nor long term treatment caused impairment of cerebral blood flow. 8-week ramipril treatment in elderly hypertensive patients was associated with improvement of some characteristics of intellectual functioning. PMID- 12891311 TI - [Effect of metoprolol CR/XL on remodeling of the heart and cardiac rhythm disturbances after myocardial infarction in patients with chronic heart failure]. AB - Metoprolol succinate was given to 60 patients with class I-III heart failure and ejection fraction <45% after myocardial infarction. Forty seven patients (mean age 59.3+/-9.8 years) completed 6 months of therapy. Dynamics of ventricular arrhythmias was assessed by Holter ECG monitoring, registration of parameters of high resolution ECG and heart rate variability. and echocardiography. Functional class of heart failure decreased in 93%, that of angina - in 55% of patients. Antiarrhythmic effect was achieved in 72% of patients. This was associated with diminishment of left ventricular volumes and increased ejection fraction, shortening of TotQRSF, and improvement of autonomic regulation of cardiac action. No dynamics of myocardial contractility and electrophysiological remodeling occurred in patients without antiarrhythmic effect. PMID- 12891312 TI - [Effect of drug therapy on frequency of adequate discharges of implanted cardioverter defibrillator]. AB - Effect of concomitant antiarrhythmic therapy (amiodarone, sotalol, beta adrenoblockers, calcium antagonists) on frequency of adequate discharges of implantable cardioverter defibrillators was studied in 149 patients. Amiodarone and sotalol significantly decreased frequency of life threatening arrhythmias (p=0.018) and defibrillator discharges(p=0.028). Beta-blockers and calcium antagonists did not affect significantly frequency of discharges. None of the antiarrhythmic drugs caused modification of ventricular tachycardia sufficient to prevent its recognition. These results can be utilized in recommendations of antiarrhythmic drug therapy in patients with implanted cardioverter defibrillators. PMID- 12891313 TI - [Mast cells in normal and diseased heart]. AB - Mast cells, producing different cytokines, chemokines, proteases, vasoactive substances and other mediators, are multifunction cells which play an important role in physiological reactions, as well as in pathological situations. In the human heart mast cells have been identified at the site of sarcolemma, in perivascular tissue, tunica adventitia of vessels, and also in coronary atheroma. Heart mast cells have a number of immunological and functional features that make them distinct from other mast cells, the features are believed to be due to microenvironmental influences on phenotypic and cytochemical characteristics. Mast cells play complex and considerable role in development of different pathological processes in the heart. Surface receptors to lgE (Fc epsilon RI) and anaphylotoxine G5a determine their participation in development of systemic and cardiac anaphylactic reactions. Direct activation of heart mast cells by intravenously injected substances used in therapy and diagnostics can also result in development of anaphylactoid reactions. It was shown that quantity and density of mast cells is much higher in patients with atherosclerosis, myocarditis, ischemic and dilated cardiomyopathy, than in humans without any cardiac pathology. Thus, the data allow to suggest that heart mast cells play an important role in regulation of a functional state of the myocardium both under normal and pathological conditions. PMID- 12891315 TI - Dangers of privatization. PMID- 12891314 TI - Diversity in food technology. PMID- 12891316 TI - Expert panel retreats from major restructuring in blueprint for NIH. PMID- 12891317 TI - Strategy for climate research gets cool response. PMID- 12891318 TI - Maize map sees geneticists split over choice of direction. PMID- 12891319 TI - Russian scientists face long fight for justice. PMID- 12891320 TI - Anger grows over plan to uproot Indian crop institute. PMID- 12891321 TI - US researchers fear job losses from privatization drive. PMID- 12891322 TI - Satellites aim to shake up quake predictions. PMID- 12891324 TI - US bombshell hits revamp of Russian weapons centres. PMID- 12891323 TI - Whale genetics study leaves conservationists all at sea. PMID- 12891326 TI - Palaeobiology: setting the record straight. PMID- 12891327 TI - Computing: quantum bits and silicon chips. PMID- 12891328 TI - Impact factors: letting everyone have their say. PMID- 12891329 TI - Impact factors reward and promote excellence. PMID- 12891335 TI - Radiation: a dose of the bomb. PMID- 12891336 TI - Global change: south dials north. PMID- 12891338 TI - Materials science: the road to diamond wafers. PMID- 12891337 TI - Developmental biology: how to turn inside out. PMID- 12891339 TI - Theoretical biology: safeguards and spurs. PMID- 12891340 TI - Neurobiology: motor proteins branch out. PMID- 12891341 TI - Condensed-matter physics: singular behaviour. PMID- 12891342 TI - Cell biology: moving inside membranes. PMID- 12891345 TI - Biomechanics: froghopper insects leap to new heights. PMID- 12891346 TI - Fibre optics: surgery by sunlight on live animals. PMID- 12891348 TI - Transformation and control of ultra-short pulses in dispersion-engineered photonic crystal fibres. AB - Photonic crystal fibres (PCFs) offer greatly enhanced design freedom compared to standard optical fibres. For example, they allow precise control of the chromatic dispersion (CD) profile--the frequency dependence of propagation speed--over a broad wavelength range. This permits studies of nonlinear pulse propagation in previously inaccessible parameter regimes. Here we report on spectral broadening of 100-fs pulses in PCFs with anomalously flat CD profiles. Maps of the spectral and spatio-temporal behaviour as a function of power show that dramatic conversion (to both longer and shorter wavelengths) can occur in remarkably short lengths of fibre, depending on the magnitude and shape of the CD profile. Because the PCFs used are single-mode at all wavelengths, the light always emerges in a fundamental guided mode. Excellent agreement is obtained between the experimental results and numerical solutions of the nonlinear wave equation, indicating that the underlying processes can be reliably modelled. These results show how, through appropriate choice of CD, nonlinearities can be efficiently harnessed to generate laser light at new wavelengths. PMID- 12891349 TI - The break-up of heavy electrons at a quantum critical point. AB - The point at absolute zero where matter becomes unstable to new forms of order is called a quantum critical point (QCP). The quantum fluctuations between order and disorder that develop at this point induce profound transformations in the finite temperature electronic properties of the material. Magnetic fields are ideal for tuning a material as close as possible to a QCP, where the most intense effects of criticality can be studied. A previous study on the heavy-electron material YbRh2Si2 found that near a field-induced QCP electrons move ever more slowly and scatter off one another with ever increasing probability, as indicated by a divergence to infinity of the electron effective mass and scattering cross section. But these studies could not shed light on whether these properties were an artefact of the applied field, or a more general feature of field-free QCPs. Here we report that, when germanium-doped YbRh2Si2 is tuned away from a chemically induced QCP by magnetic fields, there is a universal behaviour in the temperature dependence of the specific heat and resistivity: the characteristic kinetic energy of electrons is directly proportional to the strength of the applied field. We infer that all ballistic motion of electrons vanishes at a QCP, forming a new class of conductor in which individual electrons decay into collective current-carrying motions of the electron fluid. PMID- 12891350 TI - Superconductivity phase diagram of Na(x)CoO2*1.3H2O. AB - The microscopic origin of superconductivity in the high-transition-temperature (high-T(c)) copper oxides remains the subject of active inquiry; several of their electronic characteristics are well established as universal to all the known materials, forming the experimental foundation that all theories must address. The most fundamental of those characteristics, for both the copper oxides and other superconductors, is the dependence of the superconducting T(c) on the degree of electronic band filling. The recent report of superconductivity near 4 K in the layered sodium cobalt oxyhydrate, Na(0.35)CoO2*1.3H2O, is of interest owing to both its triangular cobalt-oxygen lattice and its generally analogous chemical and structural relationships to the copper oxide superconductors. Here we show that the superconducting T(c) of this compound displays the same kind of behaviour on chemical doping that is observed in the high-T(c) copper oxides. Specifically, the optimal superconducting T(c) occurs in a narrow range of sodium concentrations (and therefore electron concentrations) and decreases for both underdoped and overdoped materials, as observed in the phase diagram of the copper oxide superconductors. The analogy is not perfect, however, suggesting that Na(x)CoO2*1.3H2O, with its triangular lattice geometry and special magnetic characteristics, may provide insights into systems where coupled charge and spin dynamics play an essential role in leading to superconductivity. PMID- 12891351 TI - A recyclable catalyst that precipitates at the end of the reaction. AB - Homogeneous catalysts--which exist in the same (usually liquid) phase as reactants and products--are usually more selective than heterogeneous catalysts and far less affected by limitations due to slow transport of reactants and products, but their separation from reaction products can be costly and inefficient. This has stimulated the development of strategies that facilitate the recycling of homogeneous catalysts. Some of these methods exploit the preference of a catalyst for one of two solvents with thermoregulated miscibility; others exploit a dramatic decrease in catalyst solubility as one reagent is consumed or temperature changed after completion of the reaction. Here we describe a tungsten catalyst for the solvent-free hydrosilylation of ketones that retains its activity until essentially all of the liquid substrate is converted to liquid products, which we can then simply decant to separate the catalyst that precipitates from the products of the reaction. We attribute the ability of the catalyst to retain its solubility and hence activity until completion of the reaction to the transient formation of liquid clathrate that contains a few molecules of the substrate per molecule of the otherwise solid catalyst. Insights into the fundamental processes controlling the formation of this liquid clathrate might help to tailor other catalysts and substrates, so as to develop efficient and solvent-free schemes for reactions of practical interest. PMID- 12891352 TI - Southern Ocean origin for the resumption of Atlantic thermohaline circulation during deglaciation. AB - During the two most recent deglaciations, the Southern Hemisphere warmed before Greenland. At the same time, the northern Atlantic Ocean was exposed to meltwater discharge, which is generally assumed to reduce the formation of North Atlantic Deep Water. Yet during deglaciation, the Atlantic thermohaline circulation became more vigorous, in the transition from a weak glacial to a strong interglacial mode. Here we use a three-dimensional ocean circulation model to investigate the impact of Southern Ocean warming and the associated sea-ice retreat on the Atlantic thermohaline circulation. We find that a gradual warming in the Southern Ocean during deglaciation induces an abrupt resumption of the interglacial mode of the thermohaline circulation, triggered by increased mass transport into the Atlantic Ocean via the warm (Indian Ocean) and cold (Pacific Ocean) water route. This effect prevails over the influence of meltwater discharge, which would oppose a strengthening of the thermohaline circulation. A Southern Ocean trigger for the transition into an interglacial mode of circulation provides a consistent picture of Southern and Northern hemispheric climate change at times of deglaciation, in agreement with the available proxy records. PMID- 12891353 TI - Possible thermal and chemical stabilization of body-centred-cubic iron in the Earth's core. AB - The nature of the stable phase of iron in the Earth's solid inner core is still highly controversial. Laboratory experiments suggest the possibility of an uncharacterized phase transformation in iron at core conditions and seismological observations have indicated the possible presence of complex, inner-core layering. Theoretical studies currently suggest that the hexagonal close packed (h.c.p.) phase of iron is stable at core pressures and that the body centred cubic (b.c.c.) phase of iron becomes elastically unstable at high pressure. In other h.c.p. metals, however, a high-pressure b.c.c. form has been found to become stabilized at high temperature. We report here a quantum mechanical study of b.c.c.-iron able to model its behaviour at core temperatures as well as pressures, using ab initio molecular dynamics free-energy calculations. We find that b.c.c.-iron indeed becomes entropically stabilized at core temperatures, but in its pure state h.c.p.-iron still remains thermodynamically more favourable. The inner core, however, is not pure iron, and our calculations indicate that the b.c.c. phase will be stabilized with respect to the h.c.p. phase by sulphur or silicon impurities in the core. Consequently, a b.c.c.-structured alloy may be a strong candidate for explaining the observed seismic complexity of the inner core. PMID- 12891354 TI - Measuring fast neutrons in Hiroshima at distances relevant to atomic-bomb survivors. AB - Data from the survivors of the atomic bombs serve as the major basis for risk calculations of radiation-induced cancer in humans. A controversy has existed for almost two decades, however, concerning the possibility that neutron doses in Hiroshima may have been much larger than estimated. This controversy was based on measurements of radioisotopes activated by thermal neutrons that suggested much higher fluences at larger distances than expected. For fast neutrons, which contributed almost all the neutron dose, clear measurement validation has so far proved impossible at the large distances (900 to 1,500 m) most relevant to survivor locations. Here, the first results are reported for the detection of 63Ni produced predominantly by fast neutrons (above about 1 MeV) in copper samples from Hiroshima. This breakthrough was made possible by the development of chemical extraction methods and major improvements in the sensitivity of accelerator mass spectrometry for detection of 63Ni atoms (refs 8-11). When results are compared with 63Ni activation predicted by neutron doses for Hiroshima survivors, good agreement is observed at the distances most relevant to survivor data. These findings provide, for the first time, clear measurement validation of the neutron doses to survivors in Hiroshima. PMID- 12891355 TI - Niche lability in the evolution of a Caribbean lizard community. AB - Niche conservatism--the tendency for closely related species to be ecologically similar--is widespread. However, most studies compare closely related taxa that occur in allopatry; in sympatry, the stabilizing forces that promote niche conservatism, and thus inhibit niche shifts, may be countered by natural selection favouring ecological divergence to minimize the intensity of interspecific interactions. Consequently, the relative importance of niche conservatism versus niche divergence in determining community structure has received little attention. Here, we examine a tropical lizard community in which species have a long evolutionary history of ecological interaction. We find that evolutionary divergence overcomes niche conservatism: closely related species are no more ecologically similar than expected by random divergence and some distantly related species are ecologically similar, leading to a community in which the relationship between ecological similarity and phylogenetic relatedness is very weak. Despite this lack of niche conservatism, the ecological structuring of the community has a phylogenetic component: niche complementarity only occurs among distantly related species, which suggests that the strength of ecological interactions among species may be related to phylogeny, but it is not necessarily the most closely related species that interact most strongly. PMID- 12891356 TI - Constrained circulation at Endeavour ridge facilitates colonization by vent larvae. AB - Understanding how larvae from extant hydrothermal vent fields colonize neighbouring regions of the mid-ocean ridge system remains a major challenge in oceanic research. Among the factors considered important in the recruitment of deep-sea larvae are metabolic lifespan, the connectivity of the seafloor topography, and the characteristics of the currents. Here we use current velocity measurements from Endeavour ridge to examine the role of topographically constrained circulation on larval transport along-ridge. We show that the dominant tidal and wind-generated currents in the region are strongly attenuated within the rift valley that splits the ridge crest, and that hydrothermal plumes rising from vent fields in the valley drive a steady near-bottom inflow within the valley. Extrapolation of these findings suggests that the suppression of oscillatory currents within rift valleys of mid-ocean ridges shields larvae from cross-axis dispersal into the inhospitable deep ocean. This effect, augmented by plume-driven circulation within rift valleys having active hydrothermal venting, helps retain larvae near their source. Larvae are then exported preferentially down-ridge during regional flow events that intermittently over-ride the currents within the valley. PMID- 12891357 TI - Evolutionary capacitance as a general feature of complex gene networks. AB - An evolutionary capacitor buffers genotypic variation under normal conditions, thereby promoting the accumulation of hidden polymorphism. But it occasionally fails, thereby revealing this variation phenotypically. The principal example of an evolutionary capacitor is Hsp90, a molecular chaperone that targets an important set of signal transduction proteins. Experiments in Drosophila and Arabidopsis have demonstrated three key properties of Hsp90: (1) it suppresses phenotypic variation under normal conditions and releases this variation when functionally compromised; (2) its function is overwhelmed by environmental stress; and (3) it exerts pleiotropic effects on key developmental processes. But whether these properties necessarily make Hsp90 a significant and unique facilitator of adaptation is unclear. Here we use numerical simulations of complex gene networks, as well as genome-scale expression data from yeast single gene deletion strains, to present a mechanism that extends the scope of evolutionary capacitance beyond the action of Hsp90 alone. We illustrate that most, and perhaps all, genes reveal phenotypic variation when functionally compromised, and that the availability of loss-of-function mutations accelerates adaptation to a new optimum phenotype. However, this effect does not require the mutations to be conditional on the environment. Thus, there might exist a large class of evolutionary capacitors whose effects on phenotypic variation complement the systemic, environment-induced effects of Hsp90. PMID- 12891358 TI - Organization of cell assemblies in the hippocampus. AB - Neurons can produce action potentials with high temporal precision. A fundamental issue is whether, and how, this capability is used in information processing. According to the 'cell assembly' hypothesis, transient synchrony of anatomically distributed groups of neurons underlies processing of both external sensory input and internal cognitive mechanisms. Accordingly, neuron populations should be arranged into groups whose synchrony exceeds that predicted by common modulation by sensory input. Here we find that the spike times of hippocampal pyramidal cells can be predicted more accurately by using the spike times of simultaneously recorded neurons in addition to the animals location in space. This improvement remained when the spatial prediction was refined with a spatially dependent theta phase modulation. The time window in which spike times are best predicted from simultaneous peer activity is 10-30 ms, suggesting that cell assemblies are synchronized at this timescale. Because this temporal window matches the membrane time constant of pyramidal neurons, the period of the hippocampal gamma oscillation and the time window for synaptic plasticity, we propose that cooperative activity at this timescale is optimal for information transmission and storage in cortical circuits. PMID- 12891359 TI - Role of the prolyl isomerase Pin1 in protecting against age-dependent neurodegeneration. AB - The neuropathological hallmarks of Alzheimer's disease and other tauopathies include senile plaques and/or neurofibrillary tangles. Although mouse models have been created by overexpressing specific proteins including beta-amyloid precursor protein, presenilin and tau, no model has been generated by gene knockout. Phosphorylation of tau and other proteins on serine or threonine residues preceding proline seems to precede tangle formation and neurodegeneration in Alzheimer's disease. Notably, these phospho(Ser/Thr)-Pro motifs exist in two distinct conformations, whose conversion in some proteins is catalysed by the Pin1 prolyl isomerase. Pin1 activity can directly restore the conformation and function of phosphorylated tau or it can do so indirectly by promoting its dephosphorylation, which suggests that Pin1 is involved in neurodegeneration; however, genetic evidence is lacking. Here we show that Pin1 expression is inversely correlated with predicted neuronal vulnerability and actual neurofibrillary degeneration in Alzheimer's disease. Pin1 knockout in mice causes progressive age-dependent neuropathy characterized by motor and behavioural deficits, tau hyperphosphorylation, tau filament formation and neuronal degeneration. Thus, Pin1 is pivotal in protecting against age-dependent neurodegeneration, providing insight into the pathogenesis and treatment of Alzheimer's disease and other tauopathies. PMID- 12891360 TI - Essential role for the peroxiredoxin Prdx1 in erythrocyte antioxidant defence and tumour suppression. AB - Reactive oxygen species are involved in many cellular metabolic and signalling processes and are thought to have a role in disease, particularly in carcinogenesis and ageing. We have generated mice with targeted inactivation of Prdx1, a member of the peroxiredoxin family of antioxidant enzymes. Here we show that mice lacking Prdx1 are viable and fertile but have a shortened lifespan owing to the development beginning at about 9 months of severe haemolytic anaemia and several malignant cancers, both of which are also observed at increased frequency in heterozygotes. The haemolytic anaemia is characterized by an increase in erythrocyte reactive oxygen species, leading to protein oxidation, haemoglobin instability, Heinz body formation and decreased erythrocyte lifespan. The malignancies include lymphomas, sarcomas and carcinomas, and are frequently associated with loss of Prdx1 expression in heterozygotes, which suggests that this protein functions as a tumour suppressor. Prdx1-deficient fibroblasts show decreased proliferation and increased sensitivity to oxidative DNA damage, whereas Prdx1-null mice have abnormalities in numbers, phenotype and function of natural killer cells. Our results implicate Prdx1 as an important defence against oxidants in ageing mice. PMID- 12891361 TI - Machinery for protein sorting and assembly in the mitochondrial outer membrane. AB - Mitochondria contain translocases for the transport of precursor proteins across their outer and inner membranes. It has been assumed that the translocases also mediate the sorting of proteins to their submitochondrial destination. Here we show that the mitochondrial outer membrane contains a separate sorting and assembly machinery (SAM) that operates after the translocase of the outer membrane (TOM). Mas37 forms a constituent of the SAM complex. The central role of the SAM complex in the sorting and assembly pathway of outer membrane proteins explains the various pleiotropic functions that have been ascribed to Mas37 (refs 4, 11-15). These results suggest that the TOM complex, which can transport all kinds of mitochondrial precursor proteins, is not sufficient for the correct integration of outer membrane proteins with a complicated topology, and instead transfers precursor proteins to the SAM complex. PMID- 12891362 TI - Dimers of the N-terminal domain of phytochrome B are functional in the nucleus. AB - A plant modulates its developmental processes in response to light by several informational photoreceptors such as phytochrome. Phytochrome is a dimeric chromoprotein which regulates various aspects of plant development from seed germination to flowering. Upon absorption of red light, phytochrome translocates from the cytoplasm to the nucleus, and regulates gene expression through interaction with transcription factors such as PIF3 (refs 5-7). The phytochrome polypeptide has two domains: the amino-terminal photosensory domain with a chromophore and the carboxy-terminal domain which contains signalling motifs such as a kinase domain. The latter is widely believed to transduce the signal to downstream components. Here we show that the C-terminal domain of Arabidopsis phytochrome B (phyB), which is known as the most important member of the phytochrome family, is not directly involved in signal transduction. The N terminal domain isolated from phyB, when dimerized and localized in the nucleus, triggered full phyB responses with much higher photosensitivity than the full length phyB. These findings indicate that the C-terminal domain attenuates the activity of phyB rather than positively transducing the signal. PMID- 12891363 TI - Processivity of the single-headed kinesin KIF1A through biased binding to tubulin. AB - Conventional isoforms of the motor protein kinesin behave functionally not as 'single molecules' but as 'two molecules' paired. This dimeric structure poses a barrier to solving its mechanism. To overcome this problem, we used an unconventional kinesin KIF1A (refs 5, 6) as a model molecule. KIF1A moves processively as an independent monomer, and can also work synergistically as a functional dimer. Here we show, by measuring its movement with an optical trapping system, that a single ATP hydrolysis triggers a single stepping movement of a single KIF1A monomer. The step size is distributed stochastically around multiples of 8 nm with a gaussian-like envelope and a standard deviation of 15 nm. On average, the step is directional to the microtubule's plus-end against a load force of up to 0.15 pN. As the source for this directional movement, we show that KIF1A moves to the microtubule's plus-end by approximately 3 nm on average on binding to the microtubule, presumably by preferential binding to tubulin on the plus-end side. We propose a simple physical formulation to explain the movement of KIF1A. PMID- 12891365 TI - Proteomics technologies: probing the proteome. PMID- 12891366 TI - Mass spectrometry goes mainstream. PMID- 12891367 TI - Look, no hands. PMID- 12891368 TI - When the chips are down. PMID- 12891370 TI - Redressing the balance. PMID- 12891371 TI - Life beyond synthesis. PMID- 12891372 TI - Success in an uneven market. PMID- 12891373 TI - Comment on 'CFTR gene mutations in sarcoidosis'. PMID- 12891374 TI - Computer-based recognition of dysmorphic faces. AB - Genetic syndromes often involve craniofacial malformations. We have investigated whether a computer can recognize disease-specific facial patterns in unrelated individuals. For this, 55 photographs (256 x 256 pixel) of patients with mucopolysaccharidosis type III (n=6), Cornelia de Lange (n=12), fragile X (n=12), Prader-Willi (n=12), and Williams-Beuren (n=13) syndromes were preprocessed by a Gabor wavelet transformation. By comparing the feature vectors at 32 facial nodes, 42/55 (76%) of the patients were correctly classified. In another four patients (7%), the correct and an incorrect diagnosis scored equally well. Clinical geneticists who were shown the same photographs achieved a recognition rate of 62%. Our results prove that certain syndromes are associated with a specific facial pattern and that this pattern can be described in mathematical terms. PMID- 12891375 TI - Genetic information and life insurance: a 'real' risk? AB - Public concern about genetic discrimination, particularly access to insurance following genetic testing, has been reported in the literature. This paper aims to separate myths from realities regarding genetic discrimination in life insurance and to underline the positive aspects of allowing insurers access to relevant genetic information for underwriting purposes. We present a review of the literature pertinent to discrimination in life insurance and a comparative analysis of industries guidelines. There are few reported cases in the literature of validated genetic discrimination. However, the benefits to be gained by allowing insurers access to relevant genetic data could justify fostering a more active role in the use of genetic information by insurance companies. PMID- 12891376 TI - Medical and lay attitudes towards genetic screening and testing in Finland. AB - The purpose of this study was to compare physicians', midwives' and lay people's attitudes towards genetic screening and testing to find out whether medical education and experience influence attitudes of genetic screening and testing. The study was based on comparison of answers to joint questions in three different cross-sectional postal surveys between October 1996 and April 1998 in Finland. Target groups were physicians (study base n=772, response rate 74%, including gynaecologists, paediatricians, general practitioners and clinical geneticists), midwives and public health nurses (collectively referred to as midwives in the following; n=800, response rate 79%), and lay people (n=2000, response rate 62%). Midwives were more worried about the consequences of genetic testing and stressed the autonomy of the customer more strongly than lay people did. Furthermore, professionals considered that lay peoples' expectations as regards to genetic testing are too high. Having more medical education was related to having less 'cannot say' and missing responses. Our results do not suggest that major conflicts about the direction of genetic testing and screening would arise in near future. However, different positions and interests should be considered. Reporting in public about new prospects and developments in medical genetics should pay more attention also to concerns for balancing promises and drawbacks. PMID- 12891377 TI - Multilocus statistics to uncover epistasis and heterogeneity in complex diseases: revisiting a set of multiple sclerosis data. AB - New statistics are developed to gather the contribution of many alleles at different loci to common diseases. Both inferential and descriptive statistics are included in order to uncover epistatic effects as well as heterogeneity. The problem of multiple testing is circumvented by considering a global null hypothesis. Global testing is supplemented by descriptive methods that make use of measures like odds ratio or the P-value of individually tested allele combinations. Visualization helps to reflect complex data sets. The methods described here have been scrutinized by statistical simulations, and we show that power gains can be substantial as compared to single locus statistics. Typing data of multiple sclerosis patients and controls are investigated, representing an example of larger scale information in screening candidate genes for their impact on complex diseases. New insights emerge from this data set demonstrating genetic heterogeneity and evidence for epistasis. PMID- 12891378 TI - Juvenile hemochromatosis locus maps to chromosome 1q in a French Canadian population. AB - Juvenile hemochromatosis (JH) is a rare autosomal recessive disorder that causes iron overload. In the French Canadian region of Saguenay Lac-Saint-Jean the worldwide largest cohort of JH cases has been identified. Here, we report the mapping of this large cohort of cases to the HFE2 locus on chromosome 1q. A maximum multipoint location score of 7.02 was observed with marker D1S2344. A common ancestral haplotype, showing the presence of a founder effect, was identified. The analysis of recombinants allowed us to confirm the JH candidate region. PMID- 12891379 TI - Indication of linkage and genetic heterogeneity for asthma and atopy on chromosomes 8p and 12q in 107 French EGEA families. AB - Using the sample of 107 families with at least two asthmatic siblings, as part of the EGEA study, we have investigated linkage to asthma (or atopy) and genetic heterogeneity according to the presence/absence of atopy (or asthma) using two approaches: (1) the triangle test statistic (TTS), which considers the identical by descent (IBD) distribution among affected sib-pairs discordant for another associated phenotype (eg asthmatic sib-pairs discordant for atopy) and (2) the predivided sample test (PST), which compares the IBD distribution of marker alleles between affected sib-pairs concordant and discordant for the associated phenotype. Two regions, 8p and 12q, already reported to be linked to both asthma and atopy, were examined here. A total of 20 asthmatic sib-pairs discordant for atopy and 24 atopic pairs discordant for asthma were analyzed by both TTS and PST methods and 83 pairs with atopic asthma by PST. Some evidence for linkage was observed for two markers in the 8p23.3-p23.2 region; D8S504 for asthma with genetic heterogeneity according to the presence/absence of atopy and D8S503 for atopy with genetic heterogeneity according to the presence/absence of asthma. In the 12q14.2-q21.33 region, there was also some evidence of linkage to two markers, D12S83 and D12S95, for atopy and asthma, respectively, with genetic heterogeneity according to the presence/absence of the associated trait. Provided the small distance between the two markers on either 8p (16 cM) or 12q (21 cM), it is unclear whether one or two genetic factors are involved in either region. PMID- 12891380 TI - Manic depressive illness in a founder population. AB - Manic depressive illness (MDI) segregates within a founder population originating from S, a mountain village in Southern Italy. Identity by descent of affected persons cannot be established by direct genealogical methods. A 56 000 persons family reconstruction data base encompasses the whole population of S in the 17th and 18th centuries, and part of the population of S and neighbouring villages in the 19th and 20th centuries. We selected 10 MDI probands who were members of the S population and not evident close relatives of each other. A total of 10 other MDI probands not evidently related to the S population formed a first control group. A second control group was formed with 10 not closely related persons originating within the S population. We determined the founders common ancestors to all the probands of one group but not ancestors to the probands of other groups, and computed the genetic contribution of each founder to each proband. The distance between probands in the S MDI group was calculated with respect to all the founders. In all, 17 founders present in the ascendancy of all individuals of the S MDI group are not present in the ascendancy of the control groups: MDI patients are derived from a subpopulation within S. Two S MDI probands are far from one another in respect of the 'nonspecific' founders, but are very close in respect of the specific founders: a putative MDI trait originated from specific founders of the MDI subpopulation, and hence is identical by descent in the S population. PMID- 12891381 TI - Genetic variation at the human tissue-type plasminogen activator (tPA) locus: haplotypes and analysis of association to plasma levels of tPA. AB - Tissue-type plasminogen activator (tPA) plays a key role in thrombus dissolution and plasma levels of tPA have been associated with cardiovascular disease. We have previously resequenced regulatory and coding regions of the human tPA gene (PLAT) and identified eight single-nucleotide polymorphisms (SNPs). In a small experimental study, four common variants were associated with invasively determined vascular tPA release rates. The aim of the present study was to investigate whether there is an association between genetic variants at this locus and plasma levels of tPA. To this end, 240 Swedish individuals without cardiovascular disease were typed for the eight SNPs and an Alu insertion polymorphism at the PLAT locus, as well as for a polymorphism in the plasminogen activator inhibitor type 1 (PAI-1) promoter (PAI-1 -675 4G>5G). Stepwise regression analysis, with established predictors of plasma tPA including plasma PAI-1 and genetic variants, showed that neither genotypes nor haplotypes were major contributors to plasma tPA. The results also showed that the level of linkage disequilibrium was high at the PLAT locus, as demonstrated by the fact that only three haplotypes had a frequency above 5%. In conclusion, in the present study neither genetic variation at the PLAT locus nor the PAI-1 -675 4G>5G polymorphism was strong predictors of plasma tPA levels, which suggests that variations in other genes contribute to the heritability of this phenotype. The results also show that three haplotypes at the PLAT locus accounted for nearly 90% of the chromosomes and that they could be defined by typing only two SNPs. PMID- 12891383 TI - Promoter polymorphism influences the effect of dexamethasone on transcriptional activation of the LTC4 synthase gene. AB - The molecular mechanisms of corticosteroid action in asthma are gradually being elucidated. The LTC4S gene encodes for LTC(4) synthase, the terminal enzyme in the generation of cysteinyl-leukotrienes (cys-LTs), which are key mediators in the pathogenesis of asthma. We have identified a novel promoter polymorphism in LTC4S at position -1072 (G/A) and a -444 (A/C) polymorphism has previously been reported. We hypothesised that the LTC4S gene promoter may be a potential site of regulation by corticosteroids and that genetic polymorphism may determine their effects at this locus. Using in vitro transfection of promoter-reporter constructs, dexamethasone was shown to increase transcription of LTC4S by more than 50% for the -1072G/-444A, A-C and G-C haplotype constructs (P&<0.02), but to have no effect on the A-A haplotype (P=0.27). These data identify an interesting phenomenon that requires validation in a human study examining ex vivo production of LTC(4) in cells from genotyped asthmatic and nonasthmatic subjects. The 9% of the Caucasian asthmatic population with the A-A haplotype may have genetically predetermined lower cys-LT levels in the presence of corticosteroids compared to other patients. These findings have potential implications in the evaluation of combined corticosteroid and antileukotriene therapy in asthma. PMID- 12891382 TI - Multiple primary malignancies in osteosarcoma patients. Incidence and predictive value of osteosarcoma subtype for cancer syndromes related with osteosarcoma. AB - The overall incidence of osteosarcoma is low. However, the occurrence of osteosarcoma in a setting of multiple primary tumours is not infrequent, although population-based incidence numbers are unknown. The occurrence of osteosarcoma and other malignancies is frequently related to treatment, and can also be the result of genetic predisposition as in patients with retinoblastoma, Li-Fraumeni syndrome, Werner syndrome and Rothmund-Thomson syndrome. The aim of our study is to establish the incidence of osteosarcoma associated with other malignancies in a populationwide study and to find out if these osteosarcomas have a specific subtype, that could draw attention to a genetic predisposition to malignancy. A list of all patients registered in the Dutch National Pathology Register, named PALGA, with a diagnosis of osteosarcoma between 1975 and May 2000 was retrieved. All patients with another malignancy besides osteosarcoma were selected. All patients registered in the same period with a tonsillectomy served as a control for the occurrence of malignancy in a normal population. In a second step, only osteosarcoma patients with a history of retinoblastoma or a malignancy before the age of 46 years, since these are most probable to have a hereditary cancer syndrome, were retained for further analysis. The osteosarcomas were subtyped as common, chondroblastic, fibroblastic, teleangiectatic, anaplastic, osteoclast rich or small cell. As a control for osteosarcoma subtypes the data of 570 patients entered in two studies from the European Osteosarcoma Intergroup (EORTC/MRC) were used. Of all 938 patients registered with the diagnosis of osteosarcoma, 66 had a history of multiple primary tumours. Four patients had a surface osteosarcoma, three an extraskeletal osteosarcoma and 59 had intramedullar high-grade osteosarcoma. Of this last group, one patient was known with Rothmund-Thomson syndrome, one had retinoblastoma and 30 had their malignancies before the age of 46. Of these 32 patients, 17 had osteosarcoma of the long bones. Especially women seem to be more susceptible for the development of multiple primaries. In nine patients, the histological subtype could be assessed by revision of available histological slides. All of these patients had an osteosarcoma subtype other than common as opposed to 29% in the control group of the European Osteosarcoma Intergroup. It is concluded that although the incidence of osteosarcoma is low, the occurrence of another malignancy in osteosarcoma patients is higher than in the normal population. Specifically, osteosarcoma patients have a relative risk of 2.4 (95% confidence interval 1.88 3.07) to develop another malignancy. A noncommon subtype of osteosarcoma should draw attention to a possible genetic predisposition of the patient involved. PMID- 12891384 TI - A locus for hereditary hypotrichosis localized to human chromosome 18q21.1. AB - Hereditary hypotrichosis is a rare autosomal recessive condition characterized clinically by alopecia. Three consanguineous kindreds with multiple affected individuals were ascertained from different regions of Pakistan. A novel hypotrichosis locus was mapped to a 5.5 cM region on chromosome 18q21.1. A maximum two-point LOD score of 5.25 was obtained at marker D18S36 (theta=0.0). Three genes each for desmoglein and desmocollin proteins are located in this region. The expression in epidermal desmosomes and their connection to the keratin intermediate filaments make these genes excellent candidates for recessive hypotrichosis. PMID- 12891386 TI - Possible consequences of applying guidelines to healthy women with a family history of breast cancer. AB - Possible effects of consistently applying published guidelines on healthy women with breast cancer in their family history were analysed. We investigated 1060 unrelated breast cancer patients and calculated the numbers of first-degree relatives that would be referred to a familial cancer clinic if the guidelines were consistently applied. A first-degree relative was considered a candidate for referral if she was female, without breast cancer at the moment of the interview, alive and over the age of 24. The criteria for referral were based on one Dutch and two British guidelines. According to the Dutch guideline, for one affected woman with breast cancer, 0.25 (95% CI 0.22-0.28) healthy first-degree female relatives should be offered a consultation at a familial cancer clinic (FCC). Application of the British guidelines would lead to a similar number of referrals. Of all healthy first-degree female relatives, who should be referred to an FCC, 34-37% had an index case among their family who was already known at a genetic department. If current guidelines are consistently applied, a sharp increase in referrals to FCCs may be expected. These guidelines, however, are arbitrary and only limited data are available on the efficacy of this surveillance for high-risk healthy women. PMID- 12891387 TI - [The practice of medicine in the medico-social context of the year 2000]. PMID- 12891385 TI - SCA17 caused by homozygous repeat expansion in TBP due to partial isodisomy 6. AB - An expanded polyglutamine domain in the TATA-binding protein (TBP) has been described in patients with spinocerebellar ataxia type 17 (SCA17) characterized by cerebellar ataxia associated with dementia. TBP is a general transcription initiation factor that regulates the expression of most eukaryotic genes transcribed by RNA polymerase II. SCA17, as an autosomal dominantly inherited progressive neurodegenerative disorder, is caused by heterozygous expansion of a CAG repeat coding for glutamine. Alleles with 27 to a maximum of 44 glutamine residues were found as the normal range, whereas expansions above 45 repeat units were considered pathological. Here, we present a patient with a very severe phenotype with a late onset but rapidly progressing ataxia associated with dementia and homozygous 47 glutamine residues caused by an apparent partial isodisomy 6. This extraordinary case has important implications for the insights of TBP and SCA17. The expanded polyglutamine domain in both TBP copies is not correlated with embryonic death indicating that the normal function of the protein is not disrupted by this kind of mutation but may account for the dementia seen in this patient. PMID- 12891388 TI - [Gastric cancer and "Helicobacter pylori": epidemiological evidence and perspectives for prevention]. PMID- 12891389 TI - [Child obesity in Navarra: evolution, tendency and relation between child and adult obesity. Pecna study]. AB - INTRODUCTION: Excess of weight at an adult age is linked to an increased risk of dying. Obesity during childhood has been increasing in recent years in the developed countries. The aim of this study is to determine the prevalence of obesity in an infant-youth population of Navarra and its follow up over six years. MATERIAL AND METHODS: Longitudinal study of a cohort formed by 1,164 children of both sexes of 4, 10 and 17 years of age at the start of the study, with data collection in 1987 and 1993. Obesity was calculated using the Index of Corporal Mass or Quetelet Index (QI) higher than percentile 90 for their age and sex. RESULTS: The rate of response obtained was 63.3% (n=737). Between 1987 and 1993 there was an increase of 5% in the obese population (p<0.01). There is a significant correlation (r=0.72) between the values of the QI at the start of the study and those obtained at its conclusion. Some 61.4% of the individuals belonging to the higher quintile of the sample at the start of the study continued belonging to this quintile after six years of follow up. CONCLUSIONS: There was an increase in obesity, in correspondence with the findings of similar studies. There is good "tracking" between obesity during childhood and its presence at an adult age. The strategies for prevention of obesity must start from infancy. PMID- 12891390 TI - [Program for the early detection of hyperphenylalaninemia: 11 years on]. AB - In 1980 the Neonatal Screening Program was set in motion in Navarra. One of its aims is the early detection of hyperphenylalaninemias. In 1987 the quantification of phenylalanine came to be carried out in the Clinical Chemistry Department of the Hospital of Navarra. From then until December 1997, 51,815 newborn babies have been studied, which represents 99.24% of the infants born in Navarra. The quantification of phenylalanine in dried blood spots during 1989 enabled us to obtain the reference interval for this amino acid that is contained between 69.5 +/- 0.01 micromol/L and 139.9 +/- 0.03 micromol/L. During these 11 years we detected 2 cases of phenylketonuric hyperphenylalaninemia and one case of transitory hyperphenylalaninemia. From the clinical point of view, the children are well at present, following prescription of adequate treatment, which is based on a diet with a low phenylalanine content. PMID- 12891391 TI - [Lipidic changes in subclinical hypothyroidism. Evolution according to whether or not corrections are made of TSH levels]. AB - INTRODUCTION: The repercussion of subclinical hypothyroidism and its treatment on the levels of circulating lipids has received scarce definition. With the aim of obtaining our own data for evaluating the need of treatment in this pathology, we designed a study protocol in two phases. MATERIAL AND METHODS: In the first phase we determined the plasma values of total cholesterol, HDL-cholesterol, LDL cholesterol and triglycerides, Apo A-I, Apo B-100 and Lp(a) in two groups of patients: a control group formed by 123 healthy women from a Pamplona Health Area and a second group formed by 29 women affected by subclinical hypothyroidism who were attending our surgeries. In the second phase we included 42 women with subclinical hypothyroidism, on whom we carried out a comparative longitudinal study for three months, dividing them into two groups. One that was formed by 30 women, who were treated until their levels of TSH were normalised, and another in which the women were left in subclinical hypothyroidism. In both groups we have evaluated the lipidic profile and Apo A-I, Apo B100 and Lp (a). RESULTS: 1st phase: The total cholesterol and the LD-cholesterol were significantly higher in the group with SH than in the control group. There were no differences in the HDL cholesterol nor in the triglycerides. 2nd phase: Group 1: The figures of total cholesterol, LDL-cholesterol, HDL-cholesterol, triglycerides and Lp (a) were not modified on normalisation of the levels of TSH. The Apo A-I increased significantly and the Apo B-100 showed a tendency to diminish although without significant differences. Group 2: None of the parameters changed significantly as they remained in subclinical hypothyroidism. On comparing the final parameters between groups 1 and 2 (after 3 months), only the Apo A-I and the Lp (a) increased significantly on correction of SH. CONCLUSIONS: Subclinical hypothyroidism is accompanied by a modification of the lipidic profile of an atherogenic character although its treatment, according to our data, does not achieve significant variations. PMID- 12891392 TI - [Bacterial infections in patients infected by HIV]. AB - Individuals infected by the human immuno deficiency are more prone to suffering certain bacterial infections in the course of their clinical evolution. The agents involved in these infections are: Streptococcus pneumoniae, Haemophilus influenzae, Pseudomonas aeruginosa, Salmonella spp and Campylobacter spp, which occasion an increase in morbidity and mortality. With a lower frequency, but with an equal morbidity and mortality, infections have been found caused by Rhodococcus equi, Nocardia spp and Bartonella spp. Even though all of them account for a selective impairment of immunodeficiency in one way or another, their incidence varies. Other factors such as degree of immuno suppression, habits, social and geographic living environment could be important. In general, there is a lack of chemoprophylactic strategies for their prevention. Early diagnosis and correct treatment could have important advantages for the quality of life and survival of these patients PMID- 12891393 TI - [Neonatal screening of congenital Hypothyroidism: result of the Navarra program]. AB - The incidence of congenital Hypothyroidism is estimated at one case per 3,000 4,000 newly born. It is a proven fact that early diagnosis and treatment of this problem prevents mental retardation, which has led to the design and utilisation of methods of systematic detection in the newly born. The implantation of the screening program is relatively recent and progressive in some autonomous communities. With respect to the achievements that are being obtained by the program outstanding is the shorter time with respect to initiation of the treatment, but there still remain some shortcomings such as that of carrying out a longitudinal follow up of these infants, with respect to the evaluation of the development quotient, hence the need to have available hospital psychologists to carry out these evaluations. The expectations of achieving a mental development within normality in the children affected by congenital Hypothyroidism detected by the neonatal screening is a fact confirmed by many authors, for which reason it is highly encouraging to view the results of early detection of this neonatal screening. PMID- 12891394 TI - [Outpatient rehabilitation in the Foral community of Navarra: 1972-1997]. AB - Rehabilitation care for persons with disabilities who live in zones distant from the hospital of reference represents a challenge even for the Health Services of the most advanced countries. The requirements of the principles of "Health for All in the Year 2000" of the 1981 WHO Assembly, fully assumed by Foral Law 10/1990 (integral conception of health, efficiency, equity, decentralisation, quality and humanisation in the service) have stimulated the initiatives proceeding from the Rehabilitation Service of the "Virgen del Camino" Hospital orientated towards the pragmatic application of those principles to the reality our Community. The result has been the creation of an outpatient care network with units in both the Specialties Units and the Health Centres. As in other neighbouring Communities, we are witnessing a growing increase in the demand for rehabilitation treatment; to a great extent this is due to the increasing age of the population. This must be regulated in order to guarantee access to quality rehabilitation for every person who requires it. In order to achieve the complete fulfillment of the objectives of the WHO we propose the co-ordination between levels of attention (Primary care and Specialised care), consolidation of the figure of the rehabilitation doctor as a specialised individual providing support to Primary care in the whole community, and the realisation of certain protocols for rehabilitation referral agreed upon with the Primary care doctors PMID- 12891395 TI - [Epidemiology of HIV infection and AIDS in Navarra]. AB - The incidence of AIDS in the countries of Western Europe has been declining since 1996. The new antiretrovirals delay the progression of AIDS. Navarra has a double system for the information of HIV infections and of AIDS cases. Up until December 1997, 605 cases of AIDS were notified in Navarra, 40 of which corresponded to the final year, 1997. The annual rate of incidence was 76.4 cases per million inhabitants in 1997, and Navarra occupied the tenth place by autonomous community. In 1996 and 1997, the incidence of AIDS cases diminished in Navarra with respect to previous years. At the European level, the rates of Navarra, as is the case with the rates for Spain, are higher than those observed in all of the countries of the European Union. The most frequent category of transmission is the use of intravenous drugs in 73.2% of the cases. 13.4% are due to transmission by heterosexual contact and 6.1% to homosexual practices between males. Between 1985 and December 1997, 2,239 people were diagnosed with HIV infection in Navarra. HIV infection presents two high points in the years 1987 (249 cases) and 1991 (236 cases). Starting from the year 1992 the diagnoses diminish: 210, 173, 146, 137, 95 and 82 cases in the year 1997. We have observed a decline in the number of cases amongst users of intravenous drugs and an increase of the cases of sexual transmission, especially amongst women. PMID- 12891396 TI - [Maternal education in a rural health centre. Study of a group of pregnant women]. AB - INTRODUCTION: The basic aim in carrying out this study was to gain knowledge about the behaviour and attitude of a group of expectant women who were receiving maternal education at our Health Centre in Lesaka. This is an observational, retrospective and statistical study by the Maternal Education and Preparation for Birth Program and has been carried out without interruption since the year 1990. MATERIAL AND METHODS: The number of expectant women who have formed part of this study is 200, which corresponds to the total number who received maternal education in our Centre in the years 1991 to 1995. The changes in frequency of the utilisation of services and of lactation type have been analysed with the statistics Chi squared and Pearson and Spearman's coefficients. RESULTS: The average age of the pregnant women oscillated between 24 and 35 years. In 1991, 82.1% of the women used the services of the private tocologist and 17.9% used the public service. During 1992 a new tocological service was established in the basic zone and its use was modified in successive years to approximately 50% (Pearson's r=0.15; Chi squared= 11.2; p=0.008). In 1991 natural lactation was started with 68% of the newborn, and in 1995 this was done with 79%. CONCLUSIONS: The maternal education received by the mothers is a good instrument for achieving suitable behaviour on their part towards their children PMID- 12891397 TI - [Medicine based on the evidence of the North American legal system]. PMID- 12891398 TI - [Scientific evidence, levels of care and efficiency]. PMID- 12891399 TI - [Tungiasis: traveller parasitosis]. PMID- 12891400 TI - [Obstructive bronchiolitis with organizing pneumonia. A case that started with "Constitutional syndrome"]. PMID- 12891401 TI - [The future of Primary Health Care]. PMID- 12891402 TI - [Advances and perspectives of cancer gene therapy]. PMID- 12891403 TI - [Five years experience in the treatment of urinary lithiasis with the extracorporal lithotriptor MFL-5000]. AB - We present a review of the treatments of the urinary calculations carried out in the Lithotrity Unit in its first five years of working with the Dornier lithotriptor MFL-5000. We describe the location and size of the calculations, distribution by age and sex of the patients, energy applied and time employed, and we compare our retreatment index and the number of sessions per calculation with those published by other authors. PMID- 12891404 TI - [Lipidic profile: evolution, tendency and tracking from infancy to adulthood. PECNA Study]. AB - Cardiovascular diseases start during infancy, although the clinical manifestations show themselves during adulthood. Since the year 1987 a (PECNA) study has been underway in Navarra to analyse the epidemiology of the cardiovascular risk factors of the infant-juvenile population. This paper presents the evolution of the lipidic profile from 4 to 23 years of age, the tendency from 1987 to 1993, the evolution of the prevalence of hypercholesterolaemia seric cholesterol >200 mg/dl and the tracking from infancy to adult age. Outstanding amongst the results are the differences between the lipidic profile in both sexes, as well as the observed fall in the average levels of total seric cholesterol and in the prevalence of hypercholesterolaemia. With regard to the tracking, it is concluded that between 50% and 55% of individuals belonging to the extreme quintile of the distribution of the lipidic variables persist at this level six years later. PMID- 12891405 TI - [Eccrine pore carcinoma: an infrequent skin tumour with a high incidence in Navarra]. AB - Eccrine pore carcinoma is an infrequent malign skin tumour that derives from the eccrine sweat glands, it fundamentally affects persons of advanced age, its growth is slow and it is predominantly located in the lower extremities. Its high malignancy and its potentiality for giving rise to distant metastasis makes it necessary to carry out aggressive surgical treatments with lymphadenectomies of the ganglion chains. There are no more than 300 tumours of this type described in the literature. Our study examines 9 cases detected in Navarra in the last ten years and analyses its clinical and histopathological behaviour. The chosen surgical treatment was extirpation with safety margins of 2 cm; we did not carry out prophylactic lymphadenectomies. At present the survival rate of patients is 100% and only in one of the series has a lymphatic metastasis been detected. PMID- 12891406 TI - [The study of the inflammation of the airway in bronchial asthma by cytological and chemical analysis in the fluid phase of samples taken by sputum induction]. AB - In recent years the study of samples of bronchial biopsies and bronchoalveolar lavage fluid has made it possible to show the existence of significant airway mucosal inflammation, even in cases of mild bronchial asthma. This has led to a renewed interest in the use of sputum to assess airway inflammation non invasively. Sputum induction has recently been proposed as a non-invasive alternative to bronchoscopy for the collection of airway secretions from asthmatic subjects. Sputum induction yields satisfactory samples of sputum in the great majority of asthmatic subjects, and both differential cell counts and the measurement of molecular markers of inflammation in the sputum fluid phase are feasible. In this paper we review studies where these measurements have been taken from subjects who were asthmatic, affected by other respiratory pathologies or healthy, and we assess their validity. Finally, we consider possible future clinical applications of sputum measurement of airway inflammation in asthma. PMID- 12891407 TI - [Oxidative stress and Parkinson's disease]. AB - The etiology of Parkinson's disease (PD) is still unknown. Genetic factors and environmental exposure have been suggested in the etiopathogenesis of the disease. But the cause of dopaminergic cell loss in patients with PD remains unknown. During the last decade studies of the pathogenesis of PD have centred on the oxidative damage to the substantia nigra pars compacta. An increased free radical production and an inadequate antioxidant defence system have been reported, which could contribute to the biopathology of PD. The GSH levels in the brain are decreased, and iron levels in the substantia nigra are elevated. Moreover there are changes in SOD with an increased activity in the substantia nigra. The possible implication of oxidative stress in cell loss has encouraged research into the role of certain antioxidant agents, such as dietary compounds and drugs, as protective agents against PD. PMID- 12891408 TI - [Recent tendencies and reforms of the Health systems. Internal markets with and without competition and integrated health systems]. PMID- 12891409 TI - [Evolution of mortality amongst the youth of Navarra: 1985-1995]. AB - Recent publications show that mortality rates amongst young people and adolescents in some industrialised countries have increased in recent years. The addition of new diseases such as AIDS, which principally affect the young population, to those inevitable deaths brought about through causes such as traffic accidents has increased interest in this public health problem. The number of deaths and the adjusted global mortality rates amongst men and women of 15-34 years did not increase in Navarra in the 1985-1995 period. These are situated around 70 per 100,000. Changes have taken place in the pattern of causes, similar to those observed in other industrialised areas, with an increase of deaths through overdose and AIDS, and a decline in mortality due to traffic accidents in recent years. Traffic accidents were the first cause of death amongst youths until 1993. From this year onwards deaths from AIDS became the first cause amongst women, while amongst men the number of deaths from AIDS is equal to those caused by traffic accidents. PMID- 12891410 TI - [Situation of Diseases of Compulsory Declaration (DCDs) in Navarra. 1997]. AB - The system of Diseases of Compulsory Declaration (DCDs) encompasses, until the end of 1997, the notification of 41 transmittable infectious diseases, to which are added epidemic outbreaks of any etiology and cause. In Navarra, the DCDs are declared to the Vigilance and Epidemiological Control Section of the Public Health Institute on a weekly basis by the doctors of Primary and Specialised Care. Subsequently, the information is sent to the National Centre of Epidemiology where the data from the autonomous communities is centralised. The only disease that has shown an epidemic index higher than 1.25 is varicella, with 3,817 declared cases (EI:1.68), with a clear seasonal component--onset at the end of February (week 7) and conclusion towards mid-June (week 24). Between these two dates 80% of annual cases are declared. During 1997 there were 17 cases of Meningococcal Disease, (3.27 cases per 100,000 inhabitants), identical to the number declared in 1996 and the second lowest in the last 25 years after the rate of 2.29 in 1994. With respect to the causative serogroup, on 5 occasions serogroup C was isolated and was responsible for the death of a girl aged 3 months. On 9 occasions Neisseria meningitis serogroup B was isolated, also responsible for the death of a woman of 65. On three occasions grouping was not possible. PMID- 12891412 TI - [About the guide "Help to give up smoking" ]. PMID- 12891411 TI - [Use of cryotherapy in the Rochapea Health Centre]. AB - An analysis is made of the results obtained with the application of cryotherapy to 128 lesions: 86 common warts and 42 flat papillomas, of the 203 diagnosed in the Rochapea Health Centre in the period between October 1996 and June 30th 1997. 55.28% corresponded to warts located on the upper extremities, 34.78% were located on the head and neck, 8.07% on the lower extremities and 1.24% on the thorax. The percentage of flat papillomas was 20.68%. 2.96 sessions were needed to cure each lesion, with an average length of treatment of 42 days. No cases of postcryotherapy infection were registered and there was only one retarded complication, which was the appearance of a keloid scar. We conclude from the present study that cryotherapy is a good treatment for common warts and flat papillomas. PMID- 12891413 TI - [Intrapartum sepsis and disseminated intravascular coagulation]. PMID- 12891414 TI - [Myxoma of the left auricular simulating a leukocytoclastic vasculitis]. PMID- 12891415 TI - [The need for pharmaceutical services]. PMID- 12891416 TI - [Continued medical education]. PMID- 12891417 TI - [Tolerance to interferon beta-1b amongst patients with remittent recurrent multiple sclerosis after more than one year of treatment]. AB - INTRODUCTION: Interferon beta-1b modifies the natural history of the remittent recurrent forms of multiple sclerosis. An analysis was made of its efficiency, tolerance and adverse effects on patients with over one year of treatment. MATERIAL AND METHODS: 16 patients were studied (10 women, 6 men) with ages between 19 and 51 years, incapacity scale (EDSS) 2.61 +/- 1.07 and an annual rate of outbreaks before treatment of 1.65 +/- 0.25, who received 4 MUI/48 h/s.c. of interferon beta-1b x 15 days, afterwards continuing with 8 MUI/48 h/s.c. Corticoids were associated in four cases. Biochemical controls and mass neurological evaluations were carried out as well as a watch being kept for clinical and analytical secondary effects. RESULTS: The annual rate for outbreaks was 0.68 +/- 0.29. Amongst the most frequent adverse effects the pseudoflu syndrome was notable (87%), with an average duration of 10.46 +/- 1.4 weeks, well tolerated with paracetamol. The local reactions (87%) were light and not related to the zone of injection. The patients experienced a sensation of fatigue following the injection in 50% of the cases, although in only one case did this reach a moderate intensity. Analytical alterations were found to be 43.7%, always within the margins of grade 1 of the clinical toxicity scale. With one patient developed an acute depressive syndrome, during which treatment was interrupted, later restarted at half the maintenance dosage. CONCLUSIONS: In our series, just as in other studies, pseudoflu syndrome and local reactions were the most frequent secondary effects. Both complications are light, of brief duration and well tolerated by the patients. PMID- 12891418 TI - [Effects of a dietary intervention on the nutritional status of hemodialyzed patients: differences between men and women]. AB - The growing interest in the nutritional status of dialyzed patients arises from the fact that it plays an important role in the evolution, morbidity and mortality of renal patients. In the current study the effects have been assessed of a dietary intervention lasting 6 months, based on a food exchange list, on the nutritional status of a group of 49 hemodialysis patients as well as the different effects between men and women. The initial and final evaluation of the nutritional status was performed by estimating energy and nutrient intake with a 24 hour recall, determination of body composition by anthropometry, bioelectrical impedance and infrared interactance and biochemical measurements related to the nutritional status of the patients undergoing hemodialysis. At the end of the study, a significant increase was seen in the energy and macronutrient intake, which was more prominent in females. The body composition analysis showed a favourable evolution as compared to other studies; which was better in women who maintained the muscle mass stable and increased fat mass. The biochemical profile showed an increase of the C3 complement, while other markers such as amino acid profile and IGF I remained unchanged at the end of the study, being both similar in males and females. The dietary intervention was found to be useful in the prevention of the progression of malnutrition in these patients, apparently with a more beneficial effect on females. PMID- 12891419 TI - [Pheochromocytoma and paraganglioma]. AB - Pheochromocytoma and paraganglioma are neoplasias of the chromaffin cells that manifest themselves in some 40-60% of cases by symptoms of episodic freeing of catecholamines. They are the cause of stable and malign HTA in some 0.1-1% of cases and must be discounted in a study of the same. They most frequently affect males and their incidence increases with age. The symptomatic episodes are basically characterised by high blood pressure, migraines, sweating and palpitations, although they can have other multiple manifestations and even take a silent form in upto 40-50% of cases. They are frequently associated (10%) with multiple endocrine neoplasias and other neuroectodermic syndromes. For their diagnosis it is essential to measure the catecholamines and their metabolites, both plasmatic and urinary, and to locate the tumour by means of image tests, especially magnetic resonance. In the handling of these tumours it must be born in mind that the carrying out of physical maneuvers on the tumour and the employment of drugs that free catecholamines can induce hypertensive crises. The treatment chosen is always surgical, curative in cases of benign pheochromocytoma or producing a reduction of the tumourous mass and associated symptoms in cases of malign pheochromocytoma (10%); in this latter case treatment is complemented with chemotherapy or drugs that control the tumourous symptoms. Untreated pheochromocytoma can be a mortal disease, hence the importance of its detection and early treatment. PMID- 12891420 TI - [Quality of Life of cancer patients: studies by the Oncology Department of the Hospital of Navarra in the Quality of Life Group of the EORTC]. AB - The Quality of Life of cancer patients and its assessment are of great important nowadays. They are useful in offering a treatment that is better adapted to the characteristics of the patient and the development of his/her illness. Patients have to evaluate their Quality of Life through measurement instruments. The European Organisation of Research and Treatment of Cancer-EORTC is an international body devoted to research in cancer treatment. One of its divisions is working on the study of the Quality of Life. They have developed a core questionnaire for Quality of Life measurement and modules for different types of tumour and treatment to complement this. The Oncology Department of the Hospital of Navarra has been collaborating in this group since 1992. The Department has participated in the creation or validation of the core questionnaire and different modules. It is measuring the Quality of Life in different clinical studies. All members of the department are collaborating in these studies. PMID- 12891421 TI - [A protocol of psychological treatment (cognitive-behavioural group therapy) in patients with chronic pain in Navarra]. AB - The present conceptualisation of chronic pain as a complex experience, in which sensorial, affective-emotional and cognitive aspects are associated, holds important implications for treatment, since it poses the need for including multi disciplinary therapeutic proposals within which medico-pharmaceutical and psychological treatments are integrated. In the present paper we explain in a detailed way a protocol of psychological treatment, cognitive-behavioural group therapy, which for the next two years will be carried out with patients with chronic pain in the Pain Unit of the Hospital of Navarra and in the Traumatology service of the Ubarmin Clinic. PMID- 12891422 TI - [Incidence of cancer in Navarra. 1988-92]. AB - AIM. An exposition is given of the data on the incidence of malign tumours in Navarra in the five year period 1988-92, by age and sex groups. MATERIAL AND METHODS. The collection of data of the 1988-92 five year period was carried out in an active form in the information sources. The procedures for collection, control of duplicates and processing of previous years were maintained. Both the indicators of quality and the calculations of the different rates were obtained following the recommendations proposed by the International Agency of Cancer Research (IARC). RESULTS. Between 1988 and 1992 11,608 incident cases of malign tumours were registered in Navarra. 58% were found in men. Excluding other skin cancer, the rate adjusted to the world population was of 274.1 per 100,000 for men and 182.9 per 100,000 for women. The localisations where the adjusted rates were higher for men excluding other skin cancer were lung, prostate and bladder, and for women the localisations were breast, uterine corpus and colon. Amongst men an increasing tendency was noted of cancer of the colon and bladder, and of breast cancer amongst women. The decrease of stomach cancer in both sexes was notable. CONCLUSIONS. Navarra is situated at an intermediate level of incidence with respect to other European countries. In relation to other registers was of note the high rate of larynx cancer in men and the low rate of cervical cancer in women. PMID- 12891423 TI - [Health advertising]. AB - The purpose of this work is to make a brief review of the recent history of health advertising, of the legal basis of its control, of experience accumulated and of future perspectives in this field. Reference is made to the most frequent problems relating to the content of the advertising messages and the need is underlined for the Foral Community of Navarra to be able to legislate on the health advertising referring to the Health Centres, Services and Establishments, explicitly delimiting the criteria that will guarantee fulfillment of the principles of truthfulness, exactness, transparency, loyal competition and health protection. PMID- 12891424 TI - [Dyspepsia and infection by Helicobacter pylori: the view of the digestologist]. PMID- 12891425 TI - [Reconstruction of the amputated ear by means of auricular prosthesis stabilised with implants of osteointegrated titanium]. PMID- 12891426 TI - [Virchow-Troisier ganglion. Diagnosis of malignant of unfiliated ovarian neoplasia]. PMID- 12891428 TI - [Science and power, today and tomorrow]. PMID- 12891427 TI - [Purchasing for health care]. PMID- 12891429 TI - [Retrospective analysis of a large number of patients over 70 years old, subject to percutaneous coronary angioplasty or to surgery]. AB - INTRODUCTION: The number of patients of advanced age with symptomatic coronary illness that need surgical or percutaneous revascularisation is increasing. The aim of the present paper is to gain knowledge of the evolution of patients over 70 years of age subjected to myocardial revascularisation. MATERIAL AND METHODS: We compare two groups of patients, in a non-aleatory manner, subjected to angioplasty (N=65) and to surgery (N=75). The average age was 74.12 +/- 3.5 (70 85) and 71.97 +/- 1.9 (70-79) years respectively. The number of unhealthy vessels was greater in the surgical group (1.84 +/- 0.8 vs 2.65 +/- 0.8). RESULTS: The number revascularised vessels is greater and revascularisation more complete in the surgical group (1.14 +/- 0.4 vs 2.59 +/- 0.9) and (76% vs 49%). There is no difference in mortality in both groups. The functional degree during the evolution is worse for the angioplasty group and besides more antianginous medicines are used in treatment (1.92 +/- 1.2 vs 1.26 +/- 0.8) and they are subjected more frequently to a new revascularisation. CONCLUSIONS: Both the sick rate and the mortality of the procedure was similar in both groups. Both procedures are valid therapeutic alternatives and are acceptably safe with these patients. Surgery is more complete and is carried out on a group of patients with greater coronary distress. There are no differences in life expectancy amongst the patients treated with either of the techniques. During the follow-up period the functional degree of the surgical group is better and there are more patients treated with monotherapy. PMID- 12891430 TI - [Logistical function and quality control in a hospital service]. AB - The characteristics of Logistical Function are studied, and are applied to controlling the quality of attention in a Medical service of a Hospital. Three variables were studied: number of annual admissions, average stay and DRG coefficients. The unit of time employed was the period of one year. Within the parameters that define this Function, the value K denotes the maximum capacity of yield of the Service in relation to the studied variable. The exponential factor b is the indicator of the ascending or descending course of the variable, which is qualified by the absolute value of the same, as much in one sense as in the other. It is essential, to be able to apply this procedure, that the human and material resources of the service under study do not vary during the entire period of observation. PMID- 12891431 TI - [Acute cholecystitis: analysis of the different therapeutic approaches in a General Surgery Service during the years 1994-1996]. AB - INTRODUCTION: The commonly accepted approach in the treatment of acute cholecystitis is urgent cholecystectomy. Laparoscopy has renewed interest in the option of "cooling" the inflammatory process, in order to subsequently carry out a programmed cholecystectomy through the technique mentioned. We present an analysis of the data collected on hospital stay and medico-surgical complications in both therapeutic options. The aim of this work is to reflect on the results obtained in the treatment of acute cholecystitis and to evaluate the results of the application of laparoscopic cholecystectomy in the emergency area. MATERIAL AND METHODS: 152 patients diagnosed with acute cholecystitis are studied. They are distributed in 3 groups, analysing the simple and accumulated hospital stay, the index of medical and surgical complications and mortality. RESULTS: Of the 152 patients with a diagnosis of acute cholecystitis, 91 (59.8%) were operated on in their first admission, 47 (30.9%) were treated using "cooling" of the process to be operated on in a second admission and 14 (9.3%) by means of percutaneous cholecystostomy. A total of 29 patients were readmitted, 4 for relapse of acute cholecystitis and 25 for programmed operations. Urgent conventional cholecystectomy shows a greater rate of infections. The cholecystectomy on second admission supposes a moderate increase of the accumulated average stay. 17% of these patients were operated on using open surgery. CONCLUSIONS: Urgent surgery seems the most suitable approach in acute cholecystitis. The employment of laparoscopic surgery in these cases probably reduces the rate of the medical and surgical complications due to laparotomy in infectious acute pathology, and increases the comfort of the patient PMID- 12891432 TI - [Reconstruction of a breast following mastectomy]. AB - Breast reconstruction has become an available option for most patients undergoing mastectomy. Breast reconstruction has been shown to lessen the psychological disturbances associated with mastectomy. Reconstruction with a silicone breast prosthesis is selected when the preserved tissues after subcutaneous mastectomy, total mastectomy or modified radical mastectomy, have an adequate quantity and quality for insertion of the prosthesis. Frequently these tissues are quantitatively deficient. In these cases the use of tissue expanders allows the expansion of deficient soft tissues. Anatomically shaped expanders produce breasts with a natural appearance, adequate ptosis and a well-defined submammary fold. When the skin has poor quality or has been irradiated, or when a radical mastectomy has been performed, additional autogenous tissue needs to be brought to the breast area with a flap. The two most commonly used musculocutaneous flaps are the latissimus dorsi and the transverse rectus abdominis flap (TRAM). To get a good result nipple-areola reconstruction is necessary, and it is often necessary to modify the contralateral breast. PMID- 12891433 TI - [The utilisation of questionnaires for the psycho-social evaluation of low back pain]. AB - Three types of questionnaire are presented that are used for the study of the psycho-social conditions in cases of low back pain. By means of psychological tests such as MMPI applied in the cases of low back pain, it is possible to obtain information about the personality of the patient and some characteristics of the low back pain although these are not conclusive (differentiation between organic and functional, possibilities of its becoming chronic, result of orthopaedic treatment, possibilities of litigation, experience of the process) and the association of more than one psychological test is required. The employment of pain tests makes it possible to evaluate the affective, sensitive and intensity characteristics as a control mechanism or the forms of influencing the painful experience in the personality. The patient's work activity can be evaluated by means of overall questionnaires about the work place (such as Lest or RNUR) or by questionnaires specific to the tasks, some static (postures), others dynamic (lifting loads or repetitiveness), and they should be considered as initial factors triggering off a work-related pathology. PMID- 12891434 TI - [The need for an antibiotics policy in Primary Health Care]. PMID- 12891435 TI - [State of infectious disease notification (I.D.N.'s) in Navarra. 1996]. AB - The System of Infectious Disease Notification (I.D.N.) encompasses the notification of 41 infectious transmissible diseases to which are added epidemic outbreaks of any etiology or cause. In Navarra, the I.D.N.'s are reported to the Section of Vigilance and Epidemiological Control of the Public Health Institute. A sharp increase of alimentary toxic-infections is observed due to two community outbreaks produced by Salmonella enteritidis that affected 410 persons. The incidence of pulmonary tuberculosis, 15.37 cases per 100,000 inhabitants has remained at the same levels as previous years and with lower rates than those published in neighbouring Autonomous Communities such as La Rioja and the Autonomous Community of the Basque Country. In the group of Exanthematic Diseases, an outbreak of German measles is noteworthy that affected males between the ages of 16 and 20. During the year 1996 there were 17 cases of Meningococcal Disease that meant a rate of 3.27 per 100,000 inhabitants, the second lowest rate in the last 25 years after the rate of 2.29 of 1994. With respect to the causative serogroup, serogroup C was isolated on 7 occasions, serogroup B on four occasions and on 6 it could not be grouped. Hydatidosis through reports of I.D.N.'s and active search in the hospitals remained steady, with 26 cases, remaining within the regular values of recent years. An important decline with respect to recent years was observed in cases of hepatitis A, hepatitis B and brucellosis. PMID- 12891436 TI - [State of the meningococcal disease in Navarra]. AB - The incidence of meningococcal disease in Navarra and Spain is determined for the 1991-1997 period, with a detected density of incidence of 5.46 per 100,000 in the 1987-1991 period. In Navarra the incidence in the 1991-1997 period was of 3.26 per 100,000 inhabitants, of which 1.91 per 100,000 corresponded to serogroup B, 0.77 per 100,000 corresponded to serogroup C, and a rate of 0.57 per 100,000 were non-grouped. During the year 1997 (weeks 1 to 35) the accumulated incidence in the period of MD serogroup C was of 0.38 per 100,000 (2 cases), the Ratio of Rates Navarra/Spain was 0.21. Considering that the last case of MD occurred in week 8, in an adult woman of 39 years and nonexistence of serogroup A in Navarra, it can bi concluded that there is no justification for a strategy of mass or routine vaccination with respect to meningococcus C. PMID- 12891438 TI - [An appraisal of the pros and cons of mass vaccination]. PMID- 12891437 TI - [Public Health Nursing in Spain and Navarra throughout the present century]. AB - The evolution of humanity over the course of history, political reforms and the reforms of the health services have brought the different professions related with health to take part in this evolution. The professionals of nursing have not remained outside this evolution, changing both their functions and their formation. With respect to the field of public health, it seems that the task developed by these professionals is sometimes overlooked and lost, which is why this article attempts to present the work realised by community nurses throughout this century, paying special attention to their development in Navarra and a proposal for the future. PMID- 12891439 TI - [Myoblastoma of granular cells of endobronchial localization]. PMID- 12891440 TI - [Cutaneous metastasis as a diagnostic manifestation of malign pleural mesothelioma]. PMID- 12891441 TI - [Advancing towards better health care: decisions based on scientific evidence]. PMID- 12891442 TI - [Lewy body disease: first figures on its frequency in Spain]. AB - Lewy body disease (LBD) as a separate nosologic entity causing dementia in the elderly is being firmly established. To know its prevalence and characterization, we reviewed 549 consecutive autopsied brains in our Department. The age of death was 60 years or older in 391 subjects. Immunohistochemical staining with ubiquitin antibodies facilitated the identification of LB. Their specific density was measured (number per 100xfield) following a protocol in the predilection neocortical sites, entorhinal cortex, hippocampal gyrus, diencephalon and brainstem. We assessed the clinical features according to LB findings. Twelve brains (2.1%) had neocortical LB. Nine of them were diagnosed as diffuse Lewy body disease (DLBD). One more brain had nigral and neocortical LB leading to a pathological diagnosis of PD. In the remaining 2 cases, the finding of neocortical LB seems to be either incidental or asymptomatic or preclinical. Cognitive decline was mild to moderate in all subjects which had neocortical LB in 4 or more areas. However, the density of these LB does not correlate with the severity of dementia. Dementia was associated with minor parkinsonian symptoms and psychiatric features in the most of patients with DLBD. Thirty eight cases of the 391 (9.7%) older than 60 years in these series had been clinically diagnosed as senile dementia. Using accepted neuropathologic criteria, diagnoses were AD (63.1%), DLBD (21.05%) and vascular dementia (13.1%). These observation suggest that on consultant diagnosis of senile dementia, DLBD must be taken in account. PMID- 12891443 TI - [Study of meniscal regeneration following total meniscectomy and the replacement of meniscus by different autologous plasties]. AB - In order to study meniscal regeneration, and the behaviour of different autologous plasties used as meniscal substitutes, the knees of 45 adult White New Zealand rabbits were operated on. The 45 knees were divided into three groups with fifteen knees in each group. The meniscectomy group was given the letter M, the patellar group R, and the cartilage group C. In group M a total medial meniscectomy was performed; in group R, after the total medial meniscectomy, a strip of the patellar tendon, harvested from the same rabbit, was implanted on the site of the original meniscus; in group C, after the total medial meniscectomy, a fragment of cartilage, also harvested from the same animal, was implanted on the site of the original meniscus. We reviewed 5 knees from each group 6, 12 and 24 weeks after surgery, and the newly formed menisci were histologically studied. Twenty-four weeks after surgery we found fibrocartilaginous menisci in the three groups; the metaplasiant sequence from fibrous tissue to fibrocartilage was similar in the three groups. We therefore conclude that: 1. Meniscal regeneration occurs. 2. The autografts we have used could be valid meniscal substitutes. PMID- 12891444 TI - [Administering toxins with antithyroids. Does this prevent recidivism of Graves disease? ]. AB - There are three possibilities in the treatment of Graves-Basedow disease: antithyroids drugs, sub-total thyroidectomy and I131. In the USA there is a clear preference for definitive therapy with I131 once the thyrotoxicosis has been controlled. In Europe and Japan, however, the preference is for trying a conservative treatment, in the hope of inducing a permanent remission without recourse to radical methods. The most usual conservative pattern involves starting with high doses of antithyroids which are progressively reduced over the course of one year. The high rate of recurrence obtained with this method has fired the imagination of the endocrinologists in the search for other patterns that would provide more satisfactory results. One of these alternative patterns consists of combining thyroxine with antithyroids drugs. In this paper the characteristics of this combined pattern are reviewed. Following the justification, the clinical and experimental foundations on which it is based are outlined. Thirdly, details are given of clinical experiences taken from the medical literature and, finally, our own clinical experience is described after a five years follow up. It is concluded that: 1. The combined treatment delays the appearance of recurrence but does not reduce its frequency. 2. The addition of thyroxine alone following a course of combined treatment is not justified and 3. Prospective studies are needed in which hyperthyroid patients are grouped according to the variables that can affect recurrence. PMID- 12891445 TI - [Etiology of schizophrenia: review of genes-ambient interaction]. AB - Schizophrenia is a disorder of extraordinary importance and complexity. Heterogeneity in the clinical presentation of schizophrenia is certain, which probably reflects heterogeneity in pathophysiology and etiology. Examination of various risk factors can give clues to the causes. Only genetic risk factors come anywhere near the effect size expected for a strong causal agent in schizophrenia. However, the existence of so many environmental risk factors cannot be ignored. Both psychosocial and biological environmental factors may interact with genetic vulnerability to increase the risk of schizophrenia. PMID- 12891446 TI - [Giving up the tobacco habit through the application of the Smoker's Aid Program in the Chantrea Health Centre]. AB - INTRODUCTION: This is a prospective descriptive study of intervention without aleatory assignation, carried out in the Chantrea Health Centre. Its aim is to determine the characteristics of the smokers who are taking part in two interventions by the Smokers' Aid Program (Programa de Ayuda al Fumador-PAF), and to evaluate the factors that influence the latter's success. MATERIAL AND METHODS: The characteristics of 100 smokers were analyzed. They were offered one of the two PAF interventions: Minimal Intervention 1 (MN1): they are given a support brochure in the first consultations, contacted by telephone after one month and called to consultation after 6 months to measure CO. Minimal Intervention 2 (MN2): the smoker makes 5 consultations of support in giving up smoking (+ or - nicotine patches) with the doctor, nurse or social worker indiscriminately. The factors that were influential in abandoning the habit with 65 subjects were evaluated. RESULTS: 60% of the subjects were male with an average age of 41 (DE 29). 29% showed a pathology related to tobacco, with no relation found between this and the success of the interventions. Those who chose MN2 (38%) had started tobacco consumption at an earlier age and were more dependent on nicotine. Of the 65 smokers who completed the program, 37% continued not to smoke after 6 months. Those who managed to give up smoking were of a greater average age, had spent more years smoking and belonged above all to the MN1 intervention. DISCUSSION: Notable successes are achieved if the actions are carried out by the First Aid Team. We find no significant differences between the characteristics of those who give up smoking and those who do not. Not even a serious pathology, related to tobacco, is predictive of success. The smoker's dependence on nicotine must be taken into account in the intervention. PMID- 12891447 TI - [Study of the demand for home care by the Pamplona Extrahospital Emergency Services in 1995]. AB - The present paper is a retrospective descriptive study, carried out in the Special Casualty Service (Servicio Especial de Urgencias) of Pamplona. It evaluates 8,411 cases of calls from homes, extracted from an aleatory sample stratified by day of the week and season of the year, with a N=647. The diagnoses were codified according to the ICHPPC-2 of the WONCA and the statistical study with SPSS/PC+V 4.0. The monthly average was 701 calls from homes, with a daily average of 19 (from Monday to Friday), with 38 on Sundays and holidays. With respect to distribution by time, on working days 66.5% were distributed between 15.00 and 22.00 hours, while on Sundays and holidays 51% fell between 8.00 and 13.00 hours. The overall rate of use was situated at 32.71/1,000 inhabitants and year (with extreme rates in Mendillorri=7.42 and II Ensanche=57.35). In 74% of the cases a doctor or nurse visited the home. The most frequent cause for home care are illnesses of the respiratory apparatus with 23.5%. Some 40.9% of the patients attended were over 75 years of age. The conclusions are: 1. There is a low rate of home care. 2. Those requiring such visits are of an advanced age. 3. Illnesses of the respiratory apparatus are the most frequent cause of demand. 4. The zones of most demand are those where a Health Centre has not been set up as such. PMID- 12891448 TI - [Leishmaniasis in Navarra: review of activities]. AB - It has been recently reported a new case of human leishmaniasis in Navarra. Therefore, in this paper we have reviewed in a systematic and aggregated manner the data relating to this disease that is available in the Institute of Public Health of Navarra and that concerns different activities from 1976 to 1996, the date of notification at the Centre of the first case in humans. It includes the data about the known cases, since they were reported to the Epidemiological Surveillance System, together with that obtained by active searching (a figure that is three times higher). This review also includes the actions relating to the localisation of the transmitter vector, detected in different population centres of Navarra, as well as the degree of seroprevalence in the canine population on the census of the Foral Community. The figures make it possible to conclude that this illness is not sufficiently well-known in our area and that there is an underestimation in reporting to the Epidemiological Vigilance System. It possibly shows a growing incidence, which would call for profound reflection and the development of a program for controlling the disease, with activities directed basically against the reservoir and the transmitter vector PMID- 12891449 TI - [Evaluation of the retention of dental fissure cleft sealants three years after their application to 13-14 year old schoolchildren]. AB - The application of dental cleft sealant in the infant population is an efficient measure in the prevention of occlusal caries and has been adopted by the Program of Infant Dental Attention (Programa de Atencion Dental Infantil-PADI) in Navarra. It involves placing a polymerizable resin over the furrows and clefts of the recently emerged and healthy molars. The high levels of retention of the sealant make it possible to preserve the teeth from caries, contributing to a reduction in their incidence. The present descriptive study aims to gain knowledge of and evaluate the permanence of the sealants three years after their application. The work was carried out in 1996 in the Oral Health Service of Navarra on a sample of 160 boys and girls out of a total of 1,372 aged between 16 and 17 years in the Pamplona and Estella health area, who had been treated with cleft sealant at the age of 13-14 years. We found that after 3 years there was retention of the cleft sealant in 39% of those implanted (30% with partial retention and 9% total). In the rest of the sealants (61%) retention was nil. These results differ from those described in other studies in which the percentage of retention is higher. This leads us to affirm the need for revising and improving the technique of application and for carrying out prospective studies for subsequent checks to determine the causes of low retention. PMID- 12891450 TI - [Pamplona, tobacco free-city]. PMID- 12891451 TI - [Adipose tumours of the mediastinum. Diagnosis by axial computerized tomography. On the subject of two cases]. PMID- 12891452 TI - [Visceral leishmaniasis in the immunocompetent patient]. PMID- 12891453 TI - [Not Available]. PMID- 12891454 TI - [Present and future perspective of research on AIDS]. PMID- 12891455 TI - [Epidemiology of neurosensorial deafness in the Foral Community of Navarra]. AB - Congenital deafness is a relatively frequent disorder that shows a heterogeneous aetiology. Knowledge and the control of some risk factors have decreased the incidence of acquired deafness and increased the relative importance of genetic cause. In an attempt to investigate the clinical relevance of congenital deafness and the mechanisms of prevention in our population, a clinical and genetic study of cases with neurosensorial deafness born in Navarra between 1975 and 1990 was carried out. A total of eighty-one cases were identified, giving an incidence of 0.8 per thousand. Thirty per cent of the cases showed deafness associated with defects. Cause of deafness was identified in seventy per cent of the cases with whom a detailed clinical and genetic study could be performed (n=50). Genetic factors were responsible for the disorder in more than half of these cases. The most frequent hereditary factor was of the autosomic recessive type. This was associated with a more severe form of hearing loss. PMID- 12891456 TI - [Effects of selective vegetative denervation on the urinary bladder of the adult rat]. AB - The present work is a study of the plastic capacity of the peripheral nervous system subjected to different aggressions in the case of 28 rats of the Wistar breed. They were divided into four groups: 1) sympathetic deafness (n=9); 2) parasympathetic deafness (n=9); 3) evaluation of regeneration (n=9); 4) control group (n=5). An image analyser was used to study the acetylcholinesterase (AChE) and Tyrosine Hydroxylase (TH) positive ganglionic neurones of the pelvic ganglion (GP) as well as the dorsal ganglions (GRD) compared with the control group. With group 3 a study was also made of the possible plasticity of the transacted axons using a wheat germ agglutinin conjugated-horseradish peroxidase (WGA-HRP) neurotracer. The statistical study was carried out by means of the analysis of variance (ANOVA), Fisher test and Scheffe method, with a p<0.05 taken as significant. The results show the predominant role of the pelvic nerve in the modulation of the plastic changes produced at the ganglionic level, with a lesser influence of the hypogastric nerve. Further studies are needed in order to define the specific role of each of these in the act of miction. PMID- 12891457 TI - [Tobacco and health professionals]. AB - INTRODUCTION: This work reviews current literature on the subject published in Spanish journals that study the attitudes, understanding and behaviour of health professionals concerning tobacco consumption. MATERIAL AND METHODS: 15 papers were selected that had been carried out using surveys similar to the WHO questionnaire for health professionals. RESULTS: A classification was made of the characteristics and results obtained, there has been a significant decline in the number of consumers, with no significant difference between professions. Differences are found by sex and by age, with a greater number of women smoking. This prevalence is higher in hospitals than in primary health care. CONCLUSIONS: A positive evolution in tobacco consumption amongst professionals, with figures that are lower than those for groups of the same age amongst the general population. The need to improve understanding in dealing with individuals who smoke. PMID- 12891458 TI - [Utilisation of organs proceeding from donors with heart failure: review of the pathogenesis of organic damage and the possible mechanisms of prevention]. AB - There is an evident imbalance between the number of patients awaiting a kidney transplant and the availability of organs proceeding from donors with brain death. A high number of patients die each day from heart failure, whose organs could be used for transplants if specific care is employed. Although centres do exist where these methods of extraction are established, the problems of organic damage have yet to be resolved, since one third of the organs are still lost, besides the increase in the need for early dialysis, and the number of dysfunctioning grafts two years after the transplant, when this type or organ is employed. There is increasingly detailed knowledge of the pathogenesis of organic damage following heart failure and reanimation, as well as of the damage following the conservation and reimplantation of the kidney. Knowledge of the maximum time of hot ischemic that an organ can withstand is of crucial importance if organs are not to be unduly discarded. Besides, the increasing understanding of the physiopathology of oxidative stress could make it possible for us, through the use of antioxidants, to attempt to improve the utilisation of the organs and diminish the incidence of dysfunctions and rejections. PMID- 12891459 TI - [Aims and methods of the ISAAC study (International Study of Asthma and Allergy in Childhood)]. AB - The International Study of Asthma and Allergies in Childhood (ISAAC) is a multicentric, transversal, epidemiological study for investigating the epidemiology of asthma using a standardised methodology. In the case of Spain, centres in Almeria, Barcelona, Bilbao, Cadiz, Cartagena, Castellon, Valladolid and Valencia are collaborating in the study. The aim of the study is to obtain knowledge of the prevalence and seriousness of asthma amongst children living in different cities so as to compare the results between different Spanish cities and with other countries. This will make it possible to elaborate hypotheses on the aetiology of asthma. The ISAAC protocol studies two groups: children of 6 to 7 years of age, and adolescents of 13 to 14 years. The parents of the children of 6 to 7 years were sent questionnaires on asthma, atopic dermatitis, rhinitis and conjunctivitis, while the children in the 13 to 14 year old age group received the same written questionnaire together with a video questionnaire. A minimum of 3,000 children have been studied in each centre and age group. The information obtained from the ISAAC study, besides facilitating the study of the aetiology of asthma, will provide a baseline measure for studying the world evolution of the illness. PMID- 12891460 TI - [Study of the hospitalisations cost of complications in diabetes]. AB - In all the five hospitals of Navarra Province, with a catchment area of 525,000 inhabitants, all the hospital stays during one year (from 1/9/95 to 31/8/96) have been studied. They have been classified by the 3M, PC Grouper software. The cost of stay was calculated by the RDG weight and its relative cost in each hospital. 5.57% of admissions were diabetic patients. They made up 8.3% of the stays and accounted for 8.07% of the total expenses of hospitalisation. Ischemic cardiopathy was the main complication in the length of hospitalisation and costs. Diabetes comorbidity is responsible for a large amount of hospital expenses. Preventive care may bring a decrease in its complications and hospital costs. Further studies may show the repercussions of preventive care profits in diabetes on hospital admission rates. PMID- 12891461 TI - [Influence of postural changes on the variations of blood volume in hemodialysis and reincidence post-dialysis blood volume]. AB - The aim of the present work is to study the possible increment of blood volume, in the seated as well as the lying position, and to analyse the influence of changes of posture on the changes of blood volume during the hemodialysis. 17 patients were studied, evaluating the variations of the hematocrit in the extracorporeal circuit on entry into the dialyzer, by means of a photometric system (Crit-Line Instrument, Izasa). Every 15 minutes the values of the hematocrit were taken and heir repercussion in the blood volume in different changes of posture. Initially all of the patients improved their blood volume, both seated and lying down. During the session of hemodialysis and ultrafiltration, with the patient in a seated position, an important drop in the blood volume was noticed; this was the case in the 1st, 2nd and 3rd hours. This drop improved when the patient adopted a lying position and there was a "reincidence of post-dialysis blood volume" at the end. According to these results, the supine posture improves the refill, confirming that the patients position influences the vascular refill. PMID- 12891462 TI - [Dyspepsia and infection by Helicobacter pylori. A challenge for Primary Health Care]. PMID- 12891463 TI - [Signet-ring cells carcinoma of the colon]. PMID- 12891464 TI - [Determination of the incidence of Parkinson's disease (332, ICD 9th Rev.) in Navarra]. PMID- 12891465 TI - [Nuchal translucency measurement--an effective method for early recognition of fetal disease?]. AB - BACKGROUND: In recent years, an increasing amount of experience has been collected in measuring the nuchal translucency (NT) of the fetus in early pregnancy. While all fetuses develop a measurable collection of fluid in the area of the neck between the 11th and 14th weeks of pregnancy, the fact that fetuses with chromosomal disorders, cardiac defects, and syndromal diseases, in particular, reveal an above-average incidence of increased NT has been noticed. OBJECTIVE: By processing our own patient data from the past two years, we intend to elucidate the question of whether NT measurement is a sensible investigation for the early detection of fetal problems. PATIENTS AND METHOD: NT measurements were carried out in 199 fetuses; these measurements were standardized according to the guidelines of the Foetal Medicine Foundation in London, in whose multicentric study we are participating. The patients were under the care of our prenatal diagnosis and therapy department and were referred to us from external sources. RESULTS: NT within the reference range was determined in 152 fetuses; NT exceeded the reference value in 47 fetuses. Of those fetuses with increased NT, 7 fetuses revealed a chromosomal anomaly, 1 foetus was suffering from a cardiac defect, 3 fetuses were suffering from other organ abnormalities, and 3 fetuses were determined to be suffering from syndromal disease. None of the fetuses whose NT measurements were within the reference range was discovered to be suffering from any of the above-mentioned problems. CONCLUSION: Even this relatively small group of patients reveals that NT measurement is a very effective filter for detecting certain fetal diseases during the early fetal period. PMID- 12891466 TI - [Umbilical blood pH, Apgar scores, and early neonatal mortality]. AB - BACKGROUND: For several decades Apgar scores and umbilical blood pH sampling have been routinely used in the assessment of newborns. The aim of this study was to examine the prognostic value of neonatal survival using population-based data and to put these analyses into context with international studies. MATERIAL AND METHODS: Data from 513,135 live births from the Perinatal Birth Register of Hesse, 1990 - 1999, were used. Death in the first week of life (early neonatal death) was used as endpoint of analyses. Receiver operator characteristic analyses were employed to compare the prognostic value of Apgar scores and umbilical blood pHs independently of cutoffs used. RESULTS: Scores and umbilical blood pH showed a strong association with early neonatal morality. The relative risk of early neonatal mortality in term infants with scores 0 - 3 yields a relative risk of 1193 (95 % confidence interval = 801 - 1778) compared with babies born with scores 7 - 10. Scores at 1, 5, and 10 minutes were superior in predicting early neonatal mortality compared to umbilical blood pH, independent of cutoffs used. CONCLUSIONS: Scores remain essential in the prediction of early neonatal survival. Umbilical blood pH is also strongly and significantly associated with early neonatal death, even though it is much less pronounced than the score. DISCUSSION: The poor mortality prediction of umbilical blood pH sampling in Hesse compared with umbilical blood pH sampling in a large American delivery unit with a highly standardized measuring routine may be explained by a lower proportion of extremely acidotic infants or may be attributable to a decreased proportion of or to a less unified or less reliable measurement of the umbilical blood pH in Hesse. PMID- 12891467 TI - [Changes of fibrinolysis during labour with and without aprotinin application]. AB - OBJECTIVE: The aim of this study was to determine (a) if alpha-2-antiplasmin, plasminogen and plasminogen-activator-inhibitors (PAI 1 - 4) change in the peripartal period, (b) if these changes are explainable by the detachment of the placenta and the release of coagulation factors, and (c) if aprotinin could affect postpartal fibrinolysis when given shortly before delivery. PATIENTS AND METHODS: 84 patients have been examined (41 normal deliveries, 43 cesarean sections). After informed consent and randomization, 30 of these patients were administered 1 million KIU aprotinin (Trasylol(R)) at the latest 15 minutes before delivery (15 normal deliveries, 15 cesarean sections). The results of PAI (1 - 4), alpha-2-antiplasmin and plasminogen were collected shortly before and after delivery and 30 and 120 min after detachment of the placenta. RESULTS: Normal deliveries and cesarean sections without aprotinin showed an evident decrease of PAI (1 - 4) after delivery while alpha-2-antiplasmin and plasminogen values did not change. After administration of aprotinin the activity of PAI (1 - 4) increased slightly after labour in normal deliveries and cesarean sections and then slowly decreased later. alpha-2-Antiplasmin levels increased post partum and then decreased to the original values. Plasminogen did not change. There were no significant differences between normal deliveries and cesarean sections with regard to the three parameters studied in this work. DISCUSSION AND CONCLUSION: Changes in PAI (1 - 4) and alpha-2-antiplasmin after administration of aprotinin could be the result of a minimal demand on the fibrinolytic system. Finally, aprotinin can reduce the consumption of coagulation and fibrinolysis factors induced by detachment of the placenta. This could be used in therapy and prophylactic treatment in high-risk patients (e.g., pre-eclampsia, HELLP syndrome, etc.). PMID- 12891468 TI - [Plasma VEGF levels are increased in women with severe preeclampsia or HELLP syndrome]. AB - AIM: To investigate vascular endothelial growth factor (VEGF) serum levels in severe preeclampsia (PE) and HELLP syndrome. PATIENTS AND METHODS: Serum concentrations of VEGF, progesterone, estradiol and estriol were measured in 16 patients with PE and 14 patients with HELLP syndrome and in 30 well-matched normotensive pregnant controls. Determination of VEGF was performed by a commercially available immunoassay (Quantikine(R), R&D Systeme, Wiesbaden), those of sex steroids by a radioimmunoassay (Fa. Biermann, Bad Nauheim). RESULTS: Serum VEGF levels were significantly higher in the study than in the control group (172.0 +/- 98.9 pg/ml versus 41.4 +/- 30.5 pg/ml, U-Test: p < 0.001). In patients with HELLP syndrome mean serum VEGF concentrations were increased when compared with healthy controls but serum levels were significantly lower than in patients with PE (109.2 +/- 68.5 pg/ml versus 219.0 +/- 72.9, U-Test: p < 0.05). We could demonstrate a positive correlation between VEGF and estradiol serum concentrations in the study and control patients (Spearman rang correlation: p < 0.05). CONCLUSIONS: It is concluded that patients suffering from PE or HELLP syndrome have either an increased placental expression of VEGF as a result of hypoxia or show an increased extraplacental production of this growth factor such as in maternal or fetal endothelial cells, macrophages or smooth muscle tissue. PMID- 12891469 TI - Pruritic urticarial papules and plaques of pregnancy (PUPPP)--a case report. AB - Pruritic urticarial papules and plaques of pregnancy (PUPPP) is a rare dermatosis of unknown etiology that is most frequently seen in primiparas and twin/multiple pregnancies. The prognosis is favorable. We report a case of PUPPP in a primipara and review the clinical signs, differential diagnosis, possible etiologic factors, diagnosis, and therapy. PMID- 12891470 TI - [Investigation of a fetal heart-rate pattern that shows a reduced oscillation amplitude]. AB - A fetal heart-rate pattern that has a reduced oscillation amplitude may indicate a physiological fetal dormant period but could also be an indication of fetal hypoxemia. In some rare cases such a fetal heart rate-pattern can be an indicator of cerebral or cardial fetal malformation or of an intoxication caused by sedative drugs. Our patient is a 32-year-old Para III in the phase of 29 weeks and 3 days gestation. Upon admission to the clinic, the fetal heart-rate pattern showed a reduced oscillation amplitude, and there were no signs of fetal movement. The ultrasound examination gave us no reason to suspect fetal malformation, and the results of the Doppler ultrasonography were also normal. However, although the patient had denied taking any medication at all, the results of an toxicological blood test confirmed our suspicion of benzodiazepine intoxication. Throughout the night the fetal heart-rate pattern was continuously measured, and in the early hours of the morning, after breaking down of the oxazepam medication, a normalization of the fetal heart-rate pattern was observed. This case report definitively demonstrates that Doppler ultrasonography is a valuable method for assessing any uncertainty regarding a fetal heart-rate pattern. PMID- 12891471 TI - [CMV infections of the neonate: prevalence, diagnosis, therapy]. AB - Infections with cytomegalovirus (CMV) are still a challenge for obstetricians as well as pediatricians. CMV is the most common congenital virus infection. In the case of a primary infection, the fetus of a seronegative pregnant woman is exposed to the risk of permanent organ damage. The intrauterine transmission rate in CMV-seropositive women is low but postnatal reactivation and transmission by breast milk occur more frequently, especially in premature babies. In this review we discuss the consequences of intrauterine and postpartum transmission from a neonatological point of view. With regard to the severity of the symptoms and possible long-term consequences, the time of infection with CMV is of great importance. Prevalence, diagnostic tools, and possible options for treatment or prevention differ for each mode of CMV infection. PMID- 12891472 TI - [Faulty printing will be punished by the reader]. PMID- 12891473 TI - [Health services research: subject matter and objectives of a field at the intersection of economics, medicine and ethics]. AB - In analogy to the famous saying that war is too serious to leave it to the generals, one could say that the provision of medical care is too serious and too costly to leave it to doctors. This, however.would give the impression of an incongruity which does not exist as an actual factor or as a standpoint in research on the healthcare system. Having said that, there are differences in the methods of observation used: doctors always base their observations on their experiences at the doctor/patient level. In these situations, they are influenced by the therapeutic imperative of wanting to help and their convictions do not allow them to take into account competing requirements or to feel dependent on the higher-level conditions; and the realm of research on the health care system bases its observations more or less on findings derived by means of analysing what is often highly aggregated data, be it of a medical, economic or epidemiological nature. Even though the insights gained at the macrolevel ultimately have to be applied at the microlevel of the doctor/patient relationship, there remains an internal distance from the problems of care which is conducive to proper judgement. From this distant vantage point, researchers attempt to prove the relationship between effort and yield, to indicate weak spots which individual doctors cannot recognise and to define capacity, structure and incentive conditions which guarantee maxi-mum medical care quality. There is, for instance, evidence of astonishing dependence between the medical care provided and incentives for the number of doctor/patient contacts, use of medicines, surgery rates and number of diagnoses. This evidence can be used to draw up recommendations for self-governing bodies in the health care sector and for the political sphere as to how the health care system could be enhanced to achieve a high level of efficiency and quality. PMID- 12891474 TI - [Opportunities and risks of managed care]. AB - AIM: The present paper aims at analysing and discussing managed care with its opportunities and risks. This analysis should be a further basis for a reasonable discussion concerning the implementation of managed care elements in the German Health Care system. METHOD: On the basis of an update literature research the relevant international experiences with managed care in several health care systems--especially in the United States and Switzerland--are analysed and described. Most relevant opportunities and risks of managed care are deduced from this analysis. RESULTS: The most important opportunities of managed care are the stabilisation of health care costs, an improvement of health care processes and quality and a stronger consideration of preventive measures. The possibility of choosing between several health care models and more convenient health insurance fees are opportunities for the patients. Relevant risks of managed care involve the potential withholding of medical care with the necessity of a comprehensive quality assurance and negative influences on the physicians' autonomy. Furthermore, managed care may have negative effects on the relationship between patients and physicians or between general practitioners and medical specialists. CONCLUSIONS: Managed care has proven advantages in respect of cost stabilisation and quality improvement compared with traditional health care systems. If the risks and known problems of managed care are realised and avoided, the available opportunities could be an important option to reform the German Health Care system in respect of costs and quality. PMID- 12891475 TI - [Medical counseling of public health-insurances in questions of occupational diseases in regard to economical benefits--project of cooperation between AOK Hesse and the Medical Advisory and Expertising Service Hesse]. AB - Since September 1998 exists a project of cooperation and consultation between the AOK Hesse and the Medical Advisory and Expertising Service Hesse with the aim to identify occupational diseases and to survey decisions of the Employer's Liability Insurance Association. The procedure is based on a computer-added recognition-system, a profound preparation of the single cases by the employees of the health-insurance and a very intensively carried out deliberation by Medical Doctors of occupational medicine. In a period of four and a half year 8391 cases have been reviewed of which 4859 have already been determined. An approval as occupational disease by the Employer's Liability Insurance Association has been determined in 1954 cases, in 2905 cases the acknowledgement has not been determined. Regarding the determined cases a recourse of 10,078,922.27 EUR has been realized. In regard to the invested small resources of personnel the procedure has proved itself as highly effective to discover and to assert recourses. Beside the economical aspects for the public health-insurance, other results of the project were the assurance of the entitlement to benefits of people coming down with occupational diseases or their relatives. New insights about the actual development of occupational diseases in Germany als well as their prevention can be proceeded. PMID- 12891476 TI - [Mammography screening--neglected aspects of radiation risks]. AB - It is generally assumed that the radiation risk from mass screening for breast cancer may be neglected. This is questionable in view of the high biological effectiveness of low energy X-rays used in radiography of the breasts. The hormone replacement therapy for elderly women must be considered as a further cause for unexpected high rates of radiation-induced breast cancer cases. Each normal population of women, moreover, contains a very radiosensitive subgroup: the women bearing a mutation of the gene BRCA 1 or BRCA 2. In these, repeated X ray use must be definitely avoided. PMID- 12891477 TI - ["Breathing freely" in schools--results and model approaches concerning the air quality in classrooms]. AB - Seven schools in Lower Saxony participated in a scientific re-search project measuring the concentration of carbon dioxide (CO2), indoor air temperatures and the relative air humidity in naturally ventilated classrooms on 58 days during the period of one year. The results are as follows: The hygienic reference value of 1500 ppm CO2 (in accordance with the official German regulation DIN 1946-2) was exceeded on 7 days in summer when measurements were taken (32% of all samples in summer) and on 32 days in winter (89% of all samples in winter). The mean CO2 concentration was 1316 ppm, 766 ppm and 1652 ppm in all samples (58 measuring days), summer samples (22 measuring days) and winter samples (36 measuring days), respectively. The mean indoor air temperature was 21.7 degrees C, 22.9 degrees C and 20.9 degrees C. The mean relative humidity was 50.7%, 54.3% and 48.5%. The mean number of persons per classroom (including the teacher) was 25.1, corresponding to a mean volume of 8.5 m3 for each person and a mean area of 2.9 m2 per person. A statistical association of CO2 concentration with season, outdoor temperature and the age of the windows was found. A software programme has been developed for calculating the concentration of CO2 depending on the number of persons and the ventilation rate in a classroom. A 3-level scheme based on CO2 concentration was developed to assess the air quality. The results showed that the classrooms were not ventilated regularly but depending on the outdoor temperature and the season. Deficits concerning all 3 parameters were found. It is very likely that the deficits are not limited to the schools participating in this project. Ventilation and indoor climate deficits might cause health problems and therefore lead to costly measurement programmes. To enable schools to take their own health-related measurements, measuring instruments were purchased which can be loaned to schools in Lower Saxony. PMID- 12891478 TI - [Exposition and sensitisation to indoor allergens, house dust mite allergen and cat allergens]. AB - The study examined the exposure to biological indoor air agents and their possible role for allergies and respiratory tract illnesses of children. It was conducted as a case control study (atopic vs non-atopic children) at the four surveillance public health departments in Baden-Wurttemberg in the winter season 1999/2000 and included 379 children of the fourth class. The concentrations of the house dust mite antigens Der F1, Der p1, and Der Gr2 as well as cat allergen Fel d1 were determined in the children's bedrooms on the ground and in the mattress. Specific IgE-antibodies against allergens from house dust, mites and cat were determined in the serum of the children. For mite allergens the following medians ( micro g/g) were estimated in floor dust: Der p1 = 0.6, Der f1 = 2.3, Gr2 = 0.1; in mattresses: Der p1 = 1.2, Der f1 = 3.4, Gr2 = 0.3. The median of Fel d1 in floor dust was 0.2 microg/g, in mattresses 0.1 microg/g. Sensitisation to dust mite allergen was found to be more prevalent than sensitisation to cat. The distribution of sensitisation among the cases and controls is different. Among the cases, more subjects were sensitised to dust mites (32.9 %) and cat (13.1 %). Among the controls, 17.1 % were sensitised to dust mites and 4.1 % to cat. The results showed no direct association between the prevalence of allergies or respiratory tract illnesses and the indoor concentrations of the allergens. Possible reasons for these findings are discussed. PMID- 12891479 TI - [Influence exercised by physicians on the vaccination rate]. AB - This paper deals with the question whether physicians promote vaccinations among their patients. Besides this, we analyse which determinants influence the acceptance of vaccination among physicians. The investigations are based on data of a project granted by the DFG (Deutsche Forschungsgemeinschaft; "Munich Vaccination Study 2001"). Within this study physicians as well as parents have been interviewed. The vaccination rate of the physicians' patients are influenced by attitudes of the physician and the attitudes he/she believes his/her patients have, but not so much by aspects of his/her behaviour. How many pamphlets and posters about vaccination are available in the doctors' practice or how long consultations about vaccination last on average has only marginal effects on the rate of vaccination among the doctors' patients. If it is a political aim to enlarge vaccination rates steps should concentrate on changing relevant attitudes. PMID- 12891480 TI - Venovenous versus venoarterial extracorporeal membrane oxygenation in congenital diaphragmatic hernia. AB - BACKGROUND: Extracorporeal membrane oxygenation (ECMO) has a significant role as a final rescue modality in severe respiratory failure of the newborn with congenital diaphragmatic hernia (CDH). The objective of this study was to compare the efficiency of venovenous (VV) versus venoarterial (VA) ECMO in newborns with CDH. METHODS: A retrospective report of 11 years experience (1990 through 2001) of a single center, comparing VV and VA ECMO is given. VV ECMO was the preferred rescue modality for respiratory failure unresponsive to maximal medical therapy. Only when the placement of a VV ECMO 14F catheter was not possible, VA ECMO was used. Forty-six patients met ECMO criteria; 26 were treated with VV ECMO and 19 with VA ECMO. One patient underwent conversion from VV to VA ECMO. RESULTS: Before ECMO, there was no difference between VV and VA ECMO patients in mean oxygenation index (83 v 83), mean airway pressure (18.4 v 18.9 cm H(2)O), ECMO cannulation age (28 v 20 hours), or in the percentage of patients who needed dopamine and dobutamine (100% v 100%). From November 1994, nitric oxide (NO) was available; before ECMO, 11 of 14 (79%) VV ECMO patients received NO versus 9 of 10 (90%) patients in the VA group. VV ECMO patients were larger (3.34 v 2.77 kg; P <.05) and of advanced gestational age (39.0 v 36.9 wk; P <.05) compared with VA ECMO patients. There was no significant difference between VV and VA ECMO patients in survival rate (18 of 26, 69% v 13 of 19, 68%), ECMO duration (152 v 150 hours), time of extubation (32.0 v 33.5 days), age at discharge (73 v 81 days), or incidence of short-term intracranial complications (3.8% v 10.5%) or myocardial stun (3.8% v 15.8%). CONCLUSIONS: The authors conclude that VV ECMO is as reliable as VA ECMO in newborns with CDH in severe respiratory failure who need ECMO support and who can accommodate the VV double-lumen catheter. Because of its potential advantages, VV ECMO may be the preferred ECMO method in these infants. PMID- 12891481 TI - Meso-portal bypass in children with portal vein thrombosis: rapid increase of the intrahepatic portal venous flow after direct portal hepatic reperfusion. AB - BACKGROUND/PURPOSE: In children with portal vein (PV) thrombosis, hepatopetal portal flow can be restored by an innovative surgical procedure, the meso-portal bypass (MPB), if the umbilical portion of the intrahepatic left PV and the superior mesenteric vein are patent. This is associated with resolution of symptoms related to extrahepatic portal hypertension (EHPH). However, no data are available yet on intrahepatic hemodynamic changes after MPB. The aim of this study was to evaluate morphologic adaptation and flow characteristics of the intrahepatic PV branches (ihPVb) after MPB. METHODS: Prospective follow-up Doppler scans of the ihPVb were performed at 0.5 to 1, 3 to 6 and 12 months after MPB in 13 consecutive patients (2000-2002) and compared with preoperative findings. RESULTS: Only small ihPVb were detected preoperatively on Doppler in 8 of 13 cases. Postoperatively (median follow up 12; range, 6 to 24 months), all 13 patients had patent MPB with hepatopetal flow, and ihPVb were easy to detect with satisfactory vessel diameters and flow velocities. CONCLUSIONS: The preoperatively small ihPVb increase rapidly in diameter and hepatopetal flow velocity in patients benefiting from MPB. This correlates well with progressive resolution of their symptoms related to EHPH and reflects rapid adaptation of ihPVb. PMID- 12891482 TI - Are there any structural alterations in a neovascularized jejunal segment by omentoenteropexy? A histologic and immunohistochemistry study. AB - BACKGROUND/PURPOSE: Although the bowel neovascularization by omentoenteropexy seems to be a good alternative to bowel lengthening, there were no rigorous analyses about its histologic structure and intrinsic innervation. The aim of this study was a histologic and immunohistochemical analysis of a neovascularized jejune segment (NJS) by omentoenteropexy. METHODS: An NJS was done by omentoenteropexy in 13 Wistar rats. Seven weeks afterward, the mesentery of the NJS was ligated and divided. The NJS was separated from the normal jejunum by proximal and distal complete sections. End-to-end anatomosis was done between NJS and normal jejunum to restore bowel continuity. Twenty-six rats were sham operated. At 10 weeks, specimens were taken, and H&E, S-100 protein, and neuron specific enolase staining were done. The muscle layer thickness, mucosae layer, and myenteric plexus were analyzed. The statistical tests were the Mann-Whitney test, Friedman's test, and Spearman's correlation. RESULTS: There was a significant increase in NJS diameter. The muscle layers and mucosa were greater in the study group. The average number of neurons per intestinal ganglion was significantly fewer in the NJS. CONCLUSIONS: Omentoenteropexy provoked an increase in the diameter of NJS and contributed toward a reduction of the number of neurons in the myenteric plexus of NJS. PMID- 12891483 TI - Fetal liver magnetic resonance imaging in anterior body wall defects: a study of specimens from the museum of pathology. AB - PURPOSE: The purpose of the study was to evaluate the appearance of liver anatomy in fetuses with complex anterior body wall defects (ABWD) using magnetic resonance imaging (MRI). METHODS: Nineteen specimens from the National Museum of Pathological Anatomy (Vienna) with a wide range of ABWD were studied on a 1.5-T MR scanner (T1- and T2-weighted sequences) with special focus on the liver and the relationship between the large lesion and the adjacent structures. RESULTS: The contrast of T2-weighted sequences was superior to that of the T1-weighted images. The liver was found in an intraabdominal location in 3 cases and in a completely extraabdominal location in 11 fetuses. Five fetuses had a very special configuration or position of the liver. Furthermore, the images showed an anteriorly located confluence of hepatic veins into the inferior vena cava in 5 specimens. Associated malformations of the complexly structured malformations involved the urogenital tract, the central nervous system (CNS), the cardiac system and the musculoskeletal system. Prenatal fast magnetic resonance imaging (MRI) in one fetus confirmed the liver in a both inside/outside position, whereas the diagnosis at fetal ultrasound scan (US) in this case had been uncertain. CONCLUSIONS: These results show that MRI may play an important part in antenatal diagnosis of ABWD, complementary to prenatal US. Prenatal fast MRI should be considered if fetal US yields ambiguous findings. PMID- 12891484 TI - Liver transplantation in children with cystic fibrosis: a long-term longitudinal review of a single center's experience. AB - BACKGROUND: Improved long-term survival in cystic fibrosis (CF) has led to an increased incidence of extrapulmonary complications of this disease. Of these, end-stage liver disease is a significant cause of morbidity and mortality with liver transplantation being the only effective therapy. METHODS: Records of all CF pediatric liver transplant recipients were reviewed. RESULTS: Twelve children with CF were the recipients of 16 allografts. The 1- and 5-year survival was 91.6% and 75%, respectively. There were 5 deaths at a mean interval of 6.8 +/- 6.3 years. All of these deaths were related to pulmonary disease. Pulmonary function improved or remained stable in 8 of 9 patients tested. Despite an 83% incidence of positive sputum cultures, there was only one early mortality related to pulmonary sepsis in the setting of primary liver allograft nonfunction. CONCLUSIONS: Liver transplantation is acceptable treatment for children with CF and end-stage liver disease. Long-term survival is comparable to liver transplantation performed for other indications. Although posttransplant morbidity and mortality is related to lung disease, the authors speculate that as therapeutic improvements prolong the survival in CF, it is expected that longer survival after liver transplantation in this patient population may also be anticipated. PMID- 12891485 TI - A new algorithm for primary hypospadias repair based on tip urethroplasty. AB - BACKGROUND/PURPOSE: Decision making in hypospadias repair potentially can be simplified by tubularized incised plate (TIP) urethroplasty. The authors report management and outcomes in a consecutive series of primary hypospadias repairs in which the intent was to perform TIP. METHODS: Records of 106 consecutive boys undergoing hypospadias repair by 1 surgeon were reviewed. Position of the meatus, degree and management of curvature, technical details of the urethroplasty, and postoperative complications were recorded. RESULTS: Curvature was noted in 24 (23%) of patients, but could be corrected with preservation of the urethral plate in all but 3. In another boy, the incised plate was thought "unhealthy" for tubularization. The remaining 102 underwent TIP, of whom, 75 had distal and 27 had proximal hypospadias. Complications, primarily fistulas, occurred in 14 (13%) of these patients. The other 4 boys underwent staged repairs that utilized TIP for the glanular urethra at the second operation. CONCLUSIONS: The authors found decision making was no longer determined by meatal location as in the past, but by severity of curvature and appearance of the incised urethral plate. Because severe curvature requiring plate transection or an "unhealthy" incised plate are uncommonly encountered, TIP repair can be performed for most hypospadias operations. PMID- 12891486 TI - Pediatric trauma care: an overview of pediatric trauma systems and their practices in 18 US states. AB - PURPOSE: The aim of this study was to describe the state of pediatric trauma system development in the United States in 1997 and 1998 and to characterize the hospitalization patterns of injured children in states with different types of pediatric trauma systems. The authors also investigated the impact of sociodemographic, injury, and geographic characteristics on those hospitalization patterns. METHODS: The authors combined statewide hospital discharge data on hospitalized trauma patients less than 15 years old with data from the American Hospital Association, the Area Resource File, the Office of Management and Budget, the states' Departments of Health, and the US Census. Besides conducting descriptive analyses, the authors evaluated the role of several parameters in determining the likelihood of treatment in trauma and nontrauma centers using multivariate multinomial logistic regression models. RESULTS: There were 15 states with adult and pediatric trauma designation systems; 9 of them had statewide hospital discharge data available. In these 9 states, 77% of the discharges were from trauma centers with no pediatric designation. More severely injured children and children with injuries to the head, face, thorax, and abdomen were more likely to be discharged from trauma centers, although large percentages of these children were treated in nontrauma centers. Older children and children with commercial insurance were less likely to be treated at trauma centers, even when injury severity, body region injured, and other factors were accounted for. CONCLUSIONS: Even in states with trauma systems, a large proportion of severely injured children are treated in nontrauma center facilities. PMID- 12891487 TI - Bronchoscopy for evaluation of foreign body aspiration in children. AB - PURPOSE: The aim of this study was to present the entire spectrum of pediatric bronchoscopy performed for foreign body aspiration (FBA), with emphasis on accuracy of diagnostic tools, technical aspects, and predictors of complications. METHODS: Records of patients who underwent bronchoscopy for evaluation of FBA in our unit from 1991 to 2000, inclusive, were reviewed retrospectively. Age, sex, history of FBA, symptoms, results of radiologic studies, bronchoscopy findings, complications, and outcome were recorded and statistically analyzed to find out the most accurate diagnostic tool. RESULTS: A total of 740 bronchoscopies were done in 663 children (402 boys, 261 girls) presenting with a mean age of 3.1 +/- 0.1 years. FBA was confirmed in 563 (85%) patients, whereas normal bronchoscopic findings, signs of pulmonary infection, and endobronchial mass were noted in 43 (6%), 54 (8%), and 3 (0.4%) patients, respectively. There was significant difference between patients with and without FBA with regard to presence of definite history (91% v 54%), normal physical examination findings (14% v 46%), and normal radiologic findings (13% v 31%). However, none of these parameters or their associations were found to be reliable to predict the presence of FBA or clinical complications. Presence of history was the most sensitive (91%), accurate (84%), and specific (46 %) diagnostic tool. Bronchoscopic removal of foreign bodies was succeeded in 558 (99%) children. Worsening of respiratory tract infection (n = 13), cardiac arrest (n = 6), laryngeal edema (n = 5), pneumothorax (n = 5), pneumomediastinum (n = 2), tracheal laceration (n = 2), and bronchospasm (n = 2) were the life-threatening complications observed in 21 (4%) patients with FBA and 14 (14%) patients without FBA (P <.05). There were 5 (0.8%) deaths. Of these, 2 patients presented with cardiopulmonary arrest immediately after FBA. Although foreign bodies were removed as quickly as possible, cardiac arrest was irreversible. The remaining 3 patients died of complications of bronchoscopy as irreversible cardiac arrest during bronchoscopy (n = 1) and worsening of respiratory tract infection after bronchoscopy (n = 2). CONCLUSIONS: The third year of the life carries the highest risk for FBA. There are no specific symptoms and signs to make a clear-cut differential diagnosis between FBA and respiratory tract infection. Bronchoscopy is invariably indicated on the basis of reliable history alone even when symptoms are minimal, and imaging studies are negative. Secondary bronchoscopy should be done in patients with persistent signs and symptoms to rule out overlooked organic foreign body particles or to remove persistent granulation tissue to avoid long-term complications necessitating lobectomy. The long duration of the procedure, presence of dense granulation tissue, and type of foreign body are important predictors of complications. Bronchoscopy should be regarded as an expert procedure and done with great care to avoid lethal complications. Differential diagnosis of respiratory tract infection by various diagnostic tools is of utmost important to avoid morbidity and mortality related to needless bronchoscopy. PMID- 12891488 TI - Interventional treatment of infantile hepatic hemangioendothelioma. AB - BACKGROUND/PURPOSE: Hemangioendothelioma is the most frequent liver tumor in infancy. Untreated symptomatic patients with heart failure have a high mortality rate. Symptomatic forms may request nonoperative treatment, because surgery is burdened with high risks in patients with heart failure. The authors report their experience with interventional coil occlusion of infantile hepatic hemangioendothelioma (IHE). METHODS: Four patients (age range, 2 to 146 days; mean, 53 days) suffering from IHE associated with heart failure were treated by endovascular coil occlusion of arterial feeders. Catheter intervention was performed via an arterial (n = 2) or venous (n = 2) approach. RESULTS: Signs of heart failure resolved within 2 to 8 days after occlusion in 3 patients. Tumor regression could be observed sonographically within 4 weeks postinterventionally. In 3 children, tumor size was reduced from a mean of 544 mL (65 to 1,350) to a mean of 4 mL (2 to 6); Mean systolic peak velocity in the hepatic artery was decreased from 170 cm/s (140 to 200) before occlusion to 45 cm/s (36 to 70) during follow-up. In the fourth patient, endovascular intervention could not control a rapidly progressing hemangioendotheliomatosis, and finally a liver transplantation had to be performed. CONCLUSIONS: Interventional occlusion of feeding arteries in symptomatic IHE is a safe and effective alternative to early open surgery. The efficacy of endovascular intervention in multifocal tumors seems questionable. PMID- 12891489 TI - Low-fat diet impairs postresection intestinal adaptation in a rat model of short bowel syndrome. AB - BACKGROUND: Low-fat diets (LFD) are utilized frequently in patients with short bowel syndrome (SBS). The purpose of this study was to investigate the effects of LFD on intestinal adaptation, enterocyte proliferation, and enterocyte cell death in a rat model of SBS. METHODS: Adult male Sprague-Dawley rats were divided into 3 experimental groups: Sham-NC rats underwent bowel transection and reanastomosis and were fed normal chow (NC), SBS-NC rats underwent 75% small bowel resection and were fed NC, and SBS-rats were fed a low-fat diet (SBS-LFD). Parameters of intestinal adaptation, enterocyte proliferation, and enterocyte apoptosis were determined on day 14 after operation. RESULTS: SBS-NC rats showed a significant increase (v Sham-NC) in jejunal and ileal bowel and mucosal weight, mucosal DNA and protein, villus height, and crypt depth. A significant 67% increase in crypt cell proliferation rate and 265% increase in villus enterocyte apoptosis was seen in the ileum of SBS-NC rats compared with control animals (P <.05). SBS-LFD animals showed lower ileal mucosal weight (29%; P <.05), jejunal crypt depth (20%; P <.05), and ileal villus height (21%; P <.05). A significant decrease in villus apoptosis in jejunum (74%; P <.05) and ileum (67%; P <.05) and a decrease in cell proliferation in ileum (35%; P <.05) was seen also after exposure to LFD compared with SBS-NC. CONCLUSIONS: In a rat model of SBS, early LFD appears to inhibit parameters of intestinal adaptation. A possible mechanisms for this effect may be decreased cell proliferation. Decreased enterocyte loss via apoptosis, found in this study, may reflect a reduced number of enterocyte. PMID- 12891490 TI - Fetal tissue engineering: chest wall reconstruction. AB - BACKGROUND/PURPOSE: This study was aimed at applying fetal tissue engineering to chest wall reconstruction. METHODS: Fetal lambs underwent harvest of elastic and hyaline cartilage specimens. Once expanded in vitro, fetal chondrocytes were seeded onto synthetic scaffolds, which then were placed in a bioreactor. After birth, fetal cartilage constructs (n = 10) were implanted in autologous fashion into the ribs of all lambs (n = 6) along with identical, but acellular scaffolds, as controls (n = 6). Engineered and acellular specimens were harvested for analysis at 4 to 12 weeks postimplantation. Standard histology and matrix specific staining were performed both before implantation and after harvest on all constructs. RESULTS: Regardless of the source of chondrocytes, all fetal constructs resembled hyaline cartilage, both grossly and histologically, in vitro. In vivo, engineered implants retained hyaline characteristics for up to 10 weeks after implantation but remodeled into fibrocartilage by 12 weeks postoperatively. Mononuclear inflammatory infiltrates surrounding residual PGA/PLLA polymer fibers were noted in all specimens but most prominently in the acellular controls. CONCLUSIONS: Engineered fetal cartilage can provide structural replacement for at least up to 10 weeks after autologous, postnatal implantation in the chest wall. Fetal tissue engineering may prove useful for the treatment of severe congenital chest wall defects at birth. PMID- 12891491 TI - Pelviureteric junction obstruction: how much is the extent of the upper ureter with defective innervation needing resection? AB - BACKGROUND/PURPOSE: It is well realized that Anderson-Hyne's pyeloplasty fails to improve the drainage pattern and renal functional parameters in a few cases of hydronephrosis because of pelviureteric junction (PUJ) obstruction. The purpose of this study is to delineate the lower extent of the abnormally innervated pelviureteric junction by S-100 immunohistochemistry, which requires surgical removal. METHODS: Thirty pelviureteric junction (PUJ) specimens were analyzed by histology and by S-100 immunohistochemistry after serial sectioning of the resected ureter and compared with the variables age, calyceal separation, parenchymal thickness, differential renal function, glomerular filtration rate (GFR), and the measured constricted ureteral segment on gross examination. RESULTS: No significant correlation was noted (P >.05) when the histology was analyzed to find any association with any of the variables studied. The length of the visible constricted segment ranged from 2 mm to 15 mm (mean, 5.37 mm). The abnormally innervated segment was much longer than the length of the visible constricted segment in 24 and of the same length in 5 specimens. However, in 1 case, the abnormal innervation segment was shorter than the macroscopic constriction by 3 mm. The maximum difference in length between the visible constriction and the lower limit of defective innervation was 8 mm. CONCLUSIONS: The abnormally innervated ureteral segment below the PUJ in hydronephrosis is longer than the visible constricted segment at the time of surgery, and the ureter should be excised at least 8 mm or more beyond the visible lower limit of the constricted segment in cases of Anderson-Hyne's pyeloplasty. PMID- 12891493 TI - Urachal anomalies: ultrasonography and management. AB - BACKGROUND: Fifty-six children with anomalies of the urachus remnant identified by ultrasound scan have been encountered in the authors' hospital over the last 4 years. METHODS: Twenty of these 56 cases were symptomatic urachal remnants, whereas the urachus remnants were seen incidentally by ultrasound scanning in the other 36 patients. Symptomatic cases were treated with antibiotics or observation. Then, symptomatic cases were divided into 2 groups. One group, surgical group, was treated with surgical resection of the urachal remnant. The other group, observation group, was followed up without its surgical resection. Forty-four patients, 11 cases of symptomatic urachal remnant and 33 asymptomatic cases, were followed up, excluding patients who had surgical treatment and who were lost to follow-up. RESULTS: Thirty patients underwent periodical ultrasonographic examination during follow-up. In 9 cases, including 2 symptomatic cases, urachal remnants have disappeared during the follow-up period spontaneously. No symptom had developed during follow-up from asymptomatic cases. CONCLUSIONS: The patients with asymptomatic urachal remnants do not require follow-up, and urachal remnants, especially those under 1 year of age, do not require surgical resection unless the patient has multiple episodes. PMID- 12891492 TI - Is a routine chest x-ray necessary for children after fluoroscopically assisted central venous access? AB - PURPOSE: The aim of this study was to determine in a pediatric population whether a routine chest x-ray after central venous access is necessary when the central venous catheter is placed with intraoperative fluoroscopy. METHODS: This was a retrospective review of the charts of all patients at Children's Hospital in Denver, Colorado who underwent subclavian or internal jugular central venous catheter placement from January 1, 1998 through December 31, 2001. Age, sex, primary reason for access, access site, number of venipuncture attempts, type of catheter, intraoperative fluoroscopy results, chest x-ray results, location of the tip of the catheter, and complications were analyzed. RESULTS: There were 1,039 central venous catheters placed in 824 patients, 92.6% in the subclavian vein and 7.4% in the internal jugular vein. There were 604 (58.1%) children who had both fluoroscopy and a postprocedure chest x-ray, there were 308 (29.6%) who had only fluoroscopy, there were 117 (11.3%) who had only a postprocedure chest x ray, and there were 10 (1.0%) who had neither fluoroscopy nor chest x-ray. On completion of the procedure, there were 12 (1.1%) children with misplaced central venous catheters, only 1 (0.1%) when intraoperative fluoroscopy was used. There were 17 (1.6%) complications; 9 (0.9%) were pulmonary (pneumothorax, hemothorax, or an effusion). All children with pulmonary complications experienced clinical signs and symptoms suggestive of the complication after their central venous catheter insertion but before their postprocedure chest x-ray. CONCLUSIONS: The number of complications encountered in children who had central venous access of the subclavian vein or internal jugular central vein with intraoperative fluoroscopy was infrequent, the number of misplaced catheters was minimized with intraoperative fluoroscopy, and all children with pulmonary complications showed clinical signs suggestive of pulmonary complications before postoperative chest x ray. Therefore, children who have had central venous access of the subclavian and internal jugular vein with intraoperative fluoroscopy do not appear to require a routine chest x-ray after catheter placement unless clinical suspicion of a complication exists. PMID- 12891494 TI - Swenson revisited: a one-stage, transanal pull-through procedure for Hirschsprung's disease. AB - BACKGROUND: Hirschsprung's disease is a common cause of neonatal intestinal obstruction. A variety of operations have been used to treat this neurogenic form of bowel obstruction. This report describes an experience with a 1-stage abdominal and transanal pull-through operation in 15 patients with rectosigmoid disease. METHODS: Fifteen infants with Hirschsprung's disease included 11 boys and 4 girls. Median age at diagnosis was 3 days, and median age of operation was 30 days. Diagnosis was confirmed by rectal biopsy. Biopsies to confirm the transition zone were performed intraabdominally through a left lower quadrant incision. A Swenson pull-through was performed via transanal dissection. RESULTS: There were no instances of anastomotic leaks, wound infections or postoperative bowel obstructions. All patients survived. Two had postoperative enterocolitis. Twelve of 15 patients did well and had 2-3 bowel motions per day. One with Down's syndrome had enterocolitis and required a colostomy. Two were lost to follow up. CONCLUSIONS: A one-stage transanal pull-through procedure in infancy is a safe alternative to staged procedures for Hirschsprung's disease. PMID- 12891495 TI - Impact of AMICAR on hemorrhagic complications of ECMO: a ten-year review. AB - PURPOSE: Preliminary studies have shown aminocaproic acid (AMICAR), an inhibitor of fibrinolysis, reduced the incidence of intracranial hemorrhage and significant surgical site bleeding in patients on extracorporeal membrane oxygenation (ECMO). The purpose of this analysis is to determine if these benefits remain when AMICAR is used in a large population. METHODS: ECMO patients from a single pediatric institution, with routine use of AMICAR for "high-risk" patients, were evaluated retrospectively from 1991 to 2001. Data including diagnosis, duration of support, significant complications, and survival were recorded. These variables were compared with those of the Extracorporeal Life Support Organization (ELSO), an international ECMO registry, using a chi2 test. P less than.05 was deemed significant. RESULTS: ECMO was used 431 times during the 10-year study period. A total of 298 patients received AMICAR, most frequently for surgical procedures. The survival rate was not statistically different in the study group when compared with the ELSO Registry (P =.06). The rate of neonatal intracranial hemorrhage was not significantly different between the 2 groups (P =.133); however, the rate of surgical site bleeding was significantly reduced in the study population (P =.005). Decrease in surgical site bleeding was particularly evident in cardiac patients (P <.001). CONCLUSIONS: In this large experience, use of AMICAR for high-risk patients on ECMO did not appear to alter the rate of neonatal intracranial hemorrhage, but did significantly reduce the incidence of surgical site bleeding. AMICAR remains a valuable tool for the prevention of hemorrhage in patients undergoing operation prior to or while on ECMO. PMID- 12891496 TI - Morphology and mucosal biochemistry of gastroschisis intestine in urine-free amniotic fluid. AB - BACKGROUND/PURPOSE: The aim of this study was to determine the role of urine exposure in gastroschisis on the pathologic and biochemical aspects. METHODS: The intestines of fetal rabbits with gastroschisis (group G), gastroschisis and urethral ligation (group GL), and normal controls (group C) were studied by measuring weigh and length, intestinal diameter and wall thickness, and thickness of each intestinal layer. Number and length of villi and villi edema were evaluated. Total protein and DNA were measured in intestinal homogenate. Lactase activity and alkaline phosphatase activity were analyzed in isolated microvilli membranes. RESULTS: Intestinal length, diameter, and wall thickness were significantly different in groups G and GL compared with C but not between groups G and GL. The same was true for the thickness of the internal muscular and serosa, villi length, and villi edema. Serosal reaction was milder in group GL than in group G, and absent in C. Total protein, lactase activity, and alkaline phosphatase activity were also significantly different in G and GL compared with C but not between G and GL. CONCLUSIONS: Urine in amniotic fluid causes an increased serosal reaction but does not account for the mucosal enzyme dysfunction. PMID- 12891497 TI - Extracorporeal life support for posttraumatic acute respiratory distress syndrome at a children's medical center. AB - BACKGROUND: Primary traumatic injury was considered previously a contraindication for institution of extracorporeal life support because of high risk for persistent or new bleeding. Published experience in adults suggests that extracorporeal membrane oxygenation (ECMO) can successfully support trauma victims with pulmonary failure. The authors reviewed their experience with the use of ECMO in pediatric and adult trauma patients with acute respiratory distress syndrome (ARDS) at a children's medical center. METHODS: ECMO Center records from 1991 through 2001 (76 children, 8 adults) were reviewed to identify all patients with a primary or secondary ICD-9 diagnostic code of posttraumatic ARDS in addition to documented trauma. RESULTS: Five children and 3 adults with traumatic injury and ARDS received ECMO support. Seven patients were injured in motor vehicle collisions; one patient suffered a gunshot wound to the chest. Patient ages ranged from 21 months to 29 years (pediatric median, 4 years; range, 21 months to 18 years). Four patients had pre-ECMO laparotomies, including 3 who required splenectomy. Four patients had liver lacerations, 3 had pulmonary contusions, and 1 had a renal contusion. Median ventilation before ECMO was 6 days (range, 2 to 10). Seven of 8 patients were placed on venovenous (VV) ECMO. Seven patients had significant bleeding on ECMO. Patients were treated with blood product replacement, epsilon-aminocaproic acid (EACA), and aprotinin infusions. Surgical intervention was not required for bleeding. Six patients received hemofiltration. Median time on ECMO was 653 hours (range, 190 to 921 hours). Six of 8 patients overall survived (75%). Four of 5 pediatric patients survived. CONCLUSIONS: Children and adults with severe posttraumatic ARDS can be treated successfully on VV extracorporeal support. Hemorrhage occurs frequently but is manageable. PMID- 12891498 TI - Treatment of Crigler-Najjar type 1 disease: relevance of early liver transplantation. AB - BACKGROUND: Crigler-Najjar syndrome type 1 (CNS1) is characterized by severe unconjugated hyperbilirubinemia from birth, caused by total failure of UDP glucuronyltransferase activity. Only orthotopic liver transplantation (OLT) offers the prospect of cure. However, because the onset of neurologic deficits is unpredictable, timing of OLT remains difficult. METHODS: In our transplant center, 3 patients underwent early OLT for CNS1. Two of them (7 yr, 12 yr) showed mild to moderate neurologic deficits only few weeks before OLT, another patient (4 yr) had no signs of bilirubin encephalopathy. All patients required extensive phototherapy to control bilirubin levels. Thus, OLT was performed shortly after the onset of neurologic symptoms or as a prophylactic procedure, respectively. RESULTS: OLT was uneventful in all recipients. One of the symptomatic patients (7 yr) completely recovered from neurologic deficits at 36 months on OLT, whereas the other patient (12 yr) significantly improved symptoms at 27 months of OLT. These patients, including the 4-year-old boy, attend school at appropriate grades now. CONCLUSIONS: Irreversible brain damage (kernicterus) may occur in the course of CNS1 disease. Because no alternative treatment options are available at this time, OLT should be performed as a preventive procedure to counteract severe CNS related complications. PMID- 12891499 TI - Colonic fistula complicating electric burns--a case report. AB - Intestinal fistula is an uncommon complication of electric burns. The authors report the case of an 11-year-old child who sustained accidental burns from a high-voltage electric current and was admitted to their hospital with full thickness burns of both the upper limbs and deep burns of the anterior abdominal wall. Staged surgery was required, and the authors were involved in the management of the burn of the anterior abdominal wall. Laparotomy was done because omentum was found prolapsing out of the abdominal wound. Three scattered areas of subserosal burn of the small bowel were noticed, which required no intervention. The rest of the viscera all were normal. During the course of his stay, he had a fecal fistula on the 19th postburn day, which was found to be from the transverse colon. Conservative management of the fistula with total parenteral nutrition for 23 days resulted in complete healing. PMID- 12891500 TI - Tissue plasminogen activator for the treatment of intraabdominal abscesses in a neonate. AB - Fibrinolytic agents have been used successfully in the management of loculated empyema; however, their use in the treatment of intraabdominal abscesses is limited. The authors describe the case of a 4-week-old girl with intraabdominal abscesses secondary to intestinal perforation that were not amenable to percutaneous drainage, but were managed successfully with intracavitary administration of tissue-plasminogen activator. This case represents the first report in a human, in which tissue-plasminogen activator was used to facilitate percutaneous drainage of an intraabdominal abscess. It is also the first time a fibrinolytic agent has been used for this purpose in a child. PMID- 12891501 TI - A child with adrenocortical carcinoma who underwent percutaneous ethanol injection therapy for liver metastasis. AB - The authors encountered a 2-year-old-girl with adrenocortical carcinoma who underwent percutaneous ethanol injection therapy (PEIT) for liver metastasis. The patient had functional adrenocortical carcinoma diagnosed and underwent excision of the tumor in the right adrenal gland. Because liver metastasis was detected 11 months after surgery, the patient underwent PEIT under general anesthesia. After the treatment, the size of the metastatic tumor was reduced with calcification and then disappeared. The patient was in a good condition 3 years, 3 months after the occurrence of liver metastasis. PMID- 12891502 TI - Gastric volvulus after laparoscopic Nissen fundoplication with gastrostomy. AB - The authors present the case report of a 13-year-old boy in whom organoaxial gastric volvulus developed four months after a laparoscopic Nissen fundoplication and placement of gastrostomy. Intraoperative findings were significant for volvulus about an axis defined by 2 fixation points, one at the fundoplication site and the other in the area of the gastrostomy tube, positioned close to the pylorus. Of the 142 pediatric cases of gastric volvulus reported to date, only 7 describe it as a complication subsequent to gastric surgery, and just one case has been reported after laparoscopic Nissen fundoplication. This case shows that fundoplication and gastrostomy, a recommended treatment for gastric volvulus, does not always preclude its development. Increasing numbers of pediatric surgeons are performing laparoscopic Nissen fundoplication procedures. The authors advise surgeons to consider this potentially fatal complication, both intraoperatively when selecting a location for gastrostomy and postoperatively when evaluating complaints. PMID- 12891503 TI - Methylene blue: dangerous dye for neonates. AB - Methylene blue is a basic dye commonly used in histologic microbiologic, and tissue staining. This report describes an instance of methylene blue toxicity in a premature neonate. PMID- 12891504 TI - Axillary lipoblastoma--tumor recurrence in the right atrium. AB - This case report describes, for the first time, vascular invasion and recurrence of a lipoblastoma 6 months after the macroscopically complete excision of the initial cervico-axillary tumour. The importance of cytogenetics in the diagnosis of lipoblastoma is emphasized, as is the need to be wary of the diagnosis of lipoma in infancy. PMID- 12891505 TI - The cremasteric reflex: a useful but imperfect sign in testicular torsion. AB - The cremaster reflex has been reported to be absent in 100% of cases of testicular torsion, making it a useful sign in this difficult diagnosis. The authors report a case of surgically confirmed testicular torsion in which the cremasteric reflex clearly was present at presentation. The background of this sign and its utility in the evaluation of the acute scrotum are discussed. PMID- 12891506 TI - Multiple intestinal atresia with apple peel syndrome: successful treatment by five end-to-end anastomoses, jejunostomy, and transanastomotic silicone stent. AB - The association of apple peel bowel with multiple intestinal atresias is a rare event. The authors report a case of a 2.200-g newborn affected by this uncommon condition. The interruptions of small bowel continuity were treated successfully with 5 end-to-end anastomoses and with the construction of 2 ostomies. A gastroduodenal silicone tube (Vygon S.A., Ecouen, France) was used as an intraluminal stent. PMID- 12891507 TI - Congenital epigastric evisceration: a case report. AB - A rare case of abdominal wall defect at the epigastric midline is presented. This newborn boy had only the greater omentum eviscerated, and no other abnormalities could be detected. This case does not seem to relate to the abdominal wall defects reported so far in the literature. This singular case contributes to enlarge the spectrum of congenital defects of the abdominal wall. PMID- 12891508 TI - The association of jejunal atresia and cystic lymphangioma in the same location. AB - Cystic lymphangiomas, which are localized on the mesentery of the intestine, are rarely seen. The presence of intestinal atresia together with cystic lymphangioma in the same location has never been described before. In this study, a premature girl weighing 2,630 g who had jejunal atresia, cystic lymphangioma localized in the mesentery of the distal atretic jejunal segment, and multiple rib anomalies is presented. The association of jejunal atresia and cystic lymphangioma may be coincidental, or jejunal atresia may be caused by an intrauterine vascular compression of volvuled cystic lymphangioma. Such an association is being reported for the first time in the literature. PMID- 12891510 TI - Pericardial cysts in children: surgical or conservative approach? AB - The authors present a case of an 11-year-old boy with a history of mild asthma and cough who underwent radiographic examination of the chest. The radiograph showed a round, discrete mass in the right cardiophrenic angle. The patient underwent thoracoscopic excision, and histologic examination found a unilocular, pericardial cyst. The possible causes of mediastinal masses and an argument for their surgical removal is presented. PMID- 12891509 TI - Clinical management and molecular cytogenetic characterization in a 45,X/46,X,idic(Yp) patient with severe hypospadia. AB - Cryptorchidism and proximal hypospadia in a newborn are highly suspicious for an intersex disorder, and proper investigations should be planned immediately after birth. In some hypospadic patients, the presence of a palpable gonad in the scrotum may induce to assign the male sex, whereas the anatomy of internal and external genitalia could be extremely complex, requiring an accurate evaluation before any definitive attribution of gender. The authors present a case of an infant, referred to the hospital for surgical treatment of a proximal hypospadia, who showed ambiguous external genitalia, absence of the right gonad, a partially dysgenetic left testis, and presence of both mullerian and wolffian structures. Cytogenetic analysis detected a mosaicism with a cell line showing an isodicentric Yp chromosome and a second one, a 45, X chromosomal complement. Because the baby had been assigned previously to male gender, he underwent a staged masculinizing correction of the genital anomalies. The authors discuss the necessity of a careful evaluation of these patients at birth by a multispecialistic team, for appropriate sex assignment and for the assessment of the risk of neoplastic degeneration. PMID- 12891511 TI - A surviving infant with sirenomelia (Mermaid syndrome) associated with absent bladder. AB - The authors report a case of a surviving infant with sirenomelia (Mermaid syndrome). The child is now 4 years of age. The authors believe that this is only the fourth reported case of an infant with sirenomelia surviving beyond the neonatal period and the first associated with absent bladder. The abnormal distal aorta shown in this case supports the theory that sirenomelia is an extreme form of caudal dysgenesis rather than occurring secondary to vascular steal. PMID- 12891512 TI - Congenital adrenal hyperplasia: surgical considerations required to repair a 46,XX patient raised as a boy. AB - 21-hydroxylase deficiency (P450 CYP21) accounts for 90% of cases of congenital adrenal hyperplasia (CAH), which is associated with abnormally low cortisol and high production of androgen precursors and is the most common cause of ambiguous genitalia. Increased androgen causes in utero virilization of the fetus, consisting of clitoral enlargement, an urogenital sinus, and labioscrotal enlargement and fusion. This is the first case in an experience covering more than 30 years, of a 46,XX patient raised as a boy. The authors report a case of a Pakistani patient born of a consanguineous union, who came to medical attention at age 3 because of severe genital ambiguity; genetic analyses showed that the child was a compound heterozygote for CAH. The surgical management of this patient consisted of (1) staged hypospadias repairs preceded by testosterone therapy, (2) creation of a bladder graft neourethra, (3) removal of mullerian structures, (4) correction of bifid prepenile scrotum, and (5) insertion of testicular prostheses. The commitment to raise a 46,XX child as a boy is a very rare event. With a series of staged complex surgical procedures and careful steroid replacement, normal secondary sexual characteristics can be achieved in these children. PMID- 12891513 TI - Metastatic testicular rhabdomyosarcoma--a report of two cases. AB - Rhabdomyosarcoma is the most common type of soft tissue sarcoma in children. The tumor spreads by local extension, to regional lymph nodes, or by distant metastases. Metastatic spread to the testicle has been rarely described. The authors describe 2 boys who were found to have intratesticular metastases after presenting with primary tumors in their extremities. The first patient, an 11 year-old boy presented with primary disease in his left foot and an enlarged testicle. Rhabdomyosarcoma was found histologically in both the foot and the testicle. A second boy 17 years of age had a primary tumor involving the left upper extremity treated with amputation, chemotherapy, and radiotherapy. A relapse was noted 2 years later in the left testicle and was treated with orchiectomy. The authors discuss the implications and the management of this rare presentation of metastatic rhabdomyosarcoma. PMID- 12891514 TI - Tumoral calcinosis of the gluteal region in a child: case report with overview of different soft-tissue calcifications. AB - Tumoral calcinosis is a distinct clinical and histologic entity that is characterized by a large deposition of calcium that resembles a neoplasm and is found predominantly in adolescents and young adults in the periarticular tissues of large joints. The authors describe the clinical presentation of a 15-year-old boy admitted at our pediatric day-surgery center for surgical management of tumoral calcinosis of the left gluteal region. Complete surgical excision is the treatment of choice. PMID- 12891515 TI - Nephrogenic adenoma of the urethra: an unusual cause of hematuria in the child. AB - The authors describe a 9-year-old boy who had an accident with his bicycle. He presented with hematuria a few weeks later, and cystoscopy results showed a polypod lesion near the veru montanum. The lesion was resected, and histologic examination showed a nephrogenic adenoma (NA), which recurred 6 years later with hematuria. NA is a rare lesion in a child's urethra and can be a source of hematuria. PMID- 12891516 TI - The urinary bladder: An extremely rare location of pediatric neuroblastoma. AB - Pediatric malignant tumors in the urinary bladder are rare with a high prevalence of rhabdomyosarcomas. A 15-month-old patient was referred to the authors' center because of a urinary bladder tumor. Imaging studies disclosed a solid pelvic mass in the dome of the bladder confirmed by a cystoscopy. Surprisingly, the biopsy done during this procedure confirmed a neuroblastoma with a favorable Shimada classification. This tumor had no bad prognostic factors. But, vessel compression and local infiltration led to delayed surgery, and neoadjuvant chemotherapy was initiated. After chemotherapy, a complete surgical resection was accomplished. Currently, this patient is in complete continuous remission. Only 5 other cases have been reported. Thus, urinary neuroblastoma seems to be a very rare pediatric tumor, but it should be considered in the differential diagnoses of urinary bladder tumor. PMID- 12891517 TI - Congenital membranous colonic stenosis--case report of an extremely rare anomaly. AB - Congenital colonic stenosis is a rare anomaly. Congenital membranous colonic stenosis is more rare. The authors experienced a case of congenital membranous colonic stenosis that was diagnosed and treated successfully. To the authors' knowledge, congenital membranous colonic stenosis has not been reported previously in the literature. PMID- 12891518 TI - Pfannenstiel incision for incarcerated inguinal hernia in neonates. AB - The authors report on 2 neonates with irreducible inguinal hernia in whom a Pfannenstiel skin incision followed by lower abdominal midline fasciotomy were used to open the abdomen during emergency operation. By using this approach, the inguinal canal could also be opened and the hernia sac ligated. Bowel resection followed by anastomosis in one case and appendectomy in the other case could be performed safely intraabdominally because of the better exposure provided by using our approach compared with an inguinal incision alone. A Pfannenstiel incision followed by a midline fasciotomy decreases the risks associated with surgical intervention by enhancing exposure and contributing to good outcome by improving wound cosmesis. PMID- 12891519 TI - Octreotide for treatment of chylothorax after repair of congenital diaphragmatic hernia. AB - Chylothorax is a recognized cause of morbidity after repair of congenital diaphragmatic hernia (CDH). Management may include prolonged hospitalization with cessation of enteral feedings, repeated aspiration, chest tube drainage, total parenteral nutrition, and introduction of a medium chain triglyceride (MCT) diet as the effusion resolves. The authors report that the successful deployment of octreotide, a somatostatin analogue, hastened resolution of a postoperative chylothorax in a newborn infant with CDH. PMID- 12891523 TI - A key role for FDA GI drug advisory committee in expedited reviews. PMID- 12891524 TI - Survey highlights myths surrounding HCV. PMID- 12891520 TI - Esophageal atresia and tracheo-esophageal fistula in a patient with Digeorge syndrome. AB - DiGeorge Syndrome (DGS) is a congenital disorder that affects the thymus, parathyroid glands, and heart and brain. Thymus involvement in DGS may vary between absence/hypoplasia of thymus to various forms of reduced T cell function. TBX1 deficiency causes a number of distinct vascular and heart defects, suggesting multiple roles in cardiovascular development, specifically, formation and growth of the pharyngeal arch arteries, growth and septation of the outflow tract of the heart, interventricular septation, and conal alignment. Here the authors describe a case of DGS presenting with severe combined immunodeficiency, esophageal atresia, and tracheoesophageal fistula (TEF). DGS is an important differential diagnosis in TEF. PMID- 12891525 TI - Discovery-based research and fishing. PMID- 12891526 TI - Image of the month. Bourne test, enterovesical fistulas. PMID- 12891527 TI - Resistance to adefovir dipivoxil therapy associated with the selection of a novel mutation in the HBV polymerase. AB - BACKGROUND & AIMS: Adefovir dipivoxil effectively inhibits both hepatitis B virus (HBV) replication and disease activity in patients with chronic hepatitis B. Resistance to treatment was not observed in 2 recent large placebo-controlled 48 week studies with this drug. The aim of this study was to characterize adefovir resistance in a patient who developed clinical and virologic evidence of breakthrough during a 96-week course of treatment. METHODS: HBV DNA was PCR amplified and sequenced. Phenotypic studies used patient-derived HBV as well as specific mutations created by site-directed mutagenesis of a HBV/baculovirus recombinant. RESULTS: Following the commencement of treatment with adefovir dipivoxil, the patient initially responded with a 2.4 log(10) decrease in serum HBV DNA and normalization of alanine aminotransaminase levels by week 16. During the second year of treatment, however, serum HBV DNA rose progressively, eventually returning to near-pretreatment levels. This increase in viral replication was associated with a marked increase in alanine aminotransferase and mild changes in bilirubin, albumin, and prothrombin time. Comparison of pretreatment and posttreatment HBV DNA by polymerase chain reaction sequencing identified a novel asparagine to threonine mutation at residue rt236 in domain D of the HBV polymerase. In vitro testing of a laboratory strain encoding the rtN236T mutation and testing of patient-derived virus confirmed that the rtN236T substitution caused a marked reduction in susceptibility to adefovir. CONCLUSIONS: The development of this novel mutation in the HBV polymerase confers resistance to adefovir dipivoxil. The patient responded to subsequent lamivudine therapy, achieving normalization of alanine aminotransferase and a significant decrease in serum HBV DNA. PMID- 12891528 TI - 6-thioguanine can cause serious liver injury in inflammatory bowel disease patients. AB - BACKGROUND & AIMS: Thioguanine (6-TG) has been studied as an alternative thiopurine in inflammatory bowel disease (IBD). Short-term safety and efficacy data were favorable. Experience with 6-TG in patients with acute lymphoblastic leukemia raised long-term safety concerns when implicated in nodular regenerative hyperplasia (NRH) of the liver and portal hypertension. The aim of this study was to describe the association between 6-TG and NRH in IBD. METHODS: Liver chemistries and complete blood counts were monitored, and patients were encouraged to undergo liver biopsy. Clinical data were collected by chart review, and associations were tested by univariate and multivariable analyses. Patients were classified based on the presence (group 1) or absence (group 2) of laboratory abnormalities. RESULTS: Laboratory abnormalities occurred in 29 of 111 patients (26%). Elevations of liver enzymes and a decrease in platelet counts (<200,000) were most commonly observed. Male gender (odds ratio, 2.9; 95% CI, 1.1 7.3; P < 0.03) and preferential 6-methylmercaptopurine production on 6 mercaptopurine/azathioprine (odds ratio, 3.0; 95% CI, 1.2-7.4; P < 0.04) were independently associated with laboratory abnormalities. No association was seen with duration of 6-TG treatment, cumulative dose, or 6-TG nucleotide levels. The median increase in alanine aminotransferase, aspartate aminotransferase, and alkaline phosphatase levels was 39, 30, and 75 U/L, respectively, in group 1, and the median decrease in platelet count was 115,000 in group 1 versus 7000 in group 2 (P < 0.001). NRH occurred in 76% of patients undergoing biopsy in group 1 and 33% in group 2. CONCLUSIONS: NRH is a common finding in 6-TG-treated patients with IBD. The progression or reversibility of NRH remains unknown. Our findings suggest that 6-TG should not be considered as therapy for patients with IBD. PMID- 12891529 TI - Virtual colonoscopy using oral contrast compared with colonoscopy for the detection of patients with colorectal polyps. AB - BACKGROUND & AIMS: Virtual colonoscopy using abdominal spiral computed tomography scanning allows total colonic evaluation with minimal invasiveness. Two dimensional images and selective 3-dimensional images of the colon are used to detect colorectal lesions. This trial used conventional colonoscopy with segmental unblinding to determine the ability of virtual colonoscopy to identify patients with colorectal lesions who need conventional colonoscopy. METHODS: We studied 205 patients with virtual colonoscopy using oral iodinated contrast preceding conventional colonoscopy. Colonic lavage was achieved with an oral sodium phosphosoda preparation and colonic distention with a carbon dioxide electronic insufflator. RESULTS: The overall sensitivity and specificity of virtual colonoscopy in identifying patients with colorectal lesions was 61.8% and 70.7%, respectively. Virtual colonoscopy was more accurate in identifying patients with lesions >/=6 mm (sensitivity 84.4% and specificity 83.1%) and those with lesions >/=10 mm (sensitivity 90% and specificity 94.6%). The negative predictive value of virtual colonoscopy was 95% for a 6-mm cutoff size and 98.9% for a 10-mm cutoff. Using a 10-mm cutoff, virtual colonoscopy precludes the need for conventional colonoscopy in 86% of patients with a 1% false-negative rate (68% with a 3.4% false-negative rate when using a 6-mm cutoff). CONCLUSIONS: Virtual colonoscopy has a high sensitivity and specificity for detecting patients with significant colorectal lesions. Its high negative predictive value may help reduce the number of negative screening colonoscopies. Further studies are needed to determine what lesion cutoff size is clinically acceptable and the appropriate interval time for repeat virtual colonoscopy when it detects lesions below this cutoff size. PMID- 12891530 TI - Prospective blinded evaluation of computed tomographic colonography for screen detection of colorectal polyps. AB - BACKGROUND & AIMS: This study used a low lesion prevalence population reflective of the screening setting to estimate the sensitivity and specificity of computerized tomographic (CT) colonography for detection of colorectal polyps. METHODS: This prospective, blinded study comprised 703 asymptomatic persons at higher-than-average risk for colorectal cancer who underwent CT colonography followed by same-day colonoscopy. Two of 3 experienced readers interpreted each CT colonography examination. RESULTS: Overall lesion prevalence for adenomas >/=1 cm in diameter was 5%. Seventy percent of all lesions were proximal to the descending colon. With colonoscopy serving as the gold standard, CT colonography detected 34%, 32%, 73%, and 63% of the 59 polyps >/=1 cm for readers 1, 2, 3, and double-reading, respectively; and 35%, 29%, 57%, and 54% of the 94 polyps 5-9 mm for readers 1, 2, 3, and double-reading, respectively. Specificity for CT colonography ranged from 95% to 98% and 86% to 95% for >1 cm and 5-9-mm polyps, respectively. Interobserver variability was high for CT colonography with kappa statistic values ranging from -0.67 to 0.89. CONCLUSIONS: In a low prevalence setting, polyp detection rates at CT colonography are well below those at colonoscopy. These rates are less than previous reports based largely on high lesion prevalence cohorts. High interobserver variability warrants further investigation but may be due to the low prevalence of polyps in this cohort and the high impact on total sensitivity of each missed polyp. Specificity, based on large numbers, is high and exhibits excellent agreement among observers. PMID- 12891531 TI - Corticosteroids and immunomodulators: postoperative infectious complication risk in inflammatory bowel disease patients. AB - BACKGROUND & AIMS: Many patients with inflammatory bowel disease receive corticosteroids and 6-mercaptopurine/azathioprine during elective bowel surgery. We investigated the postoperative infection risk for patients undergoing elective bowel surgery who were receiving corticosteroids and/or 6 mercaptopurine/azathioprine before surgery compared with patients not receiving these medications. METHODS: A retrospective cohort study was conducted on 159 patients with inflammatory bowel disease who underwent elective bowel surgery. There were 56 patients receiving corticosteroids alone, 52 patients receiving 6 mercaptopurine/azathioprine alone or with corticosteroids, and 51 patients receiving neither corticosteroids nor 6-mercaptopurine/azathioprine. Postoperative infectious complications to time of discharge were categorized into major and minor complications. RESULTS: Patients receiving corticosteroids had an adjusted odds ratio for any and major infectious complications of 3.69 (95% confidence interval [CI], 1.24-10.97) and 5.54 (95% CI, 1.12-27.26), respectively. The adjusted odds ratio for patients receiving 6 mercaptopurine/azathioprine for any and major infectious complications was 1.68 (95% CI, 0.65-4.27) and 1.20 (95% CI, 0.37-3.94), respectively. CONCLUSIONS: Preoperative use of corticosteroids in patients with inflammatory bowel disease who are undergoing elective bowel surgery is associated with an increased risk of postoperative infectious complications. 6-Mercaptopurine/azathioprine alone and the addition of 6-mercaptopurine/azathioprine for patients receiving corticosteroids was not found to significantly increase the risk of postoperative infectious complications. PMID- 12891532 TI - Kaposi of Kaposi's sarcoma. PMID- 12891533 TI - Daily soluble aspirin and prevention of colorectal adenoma recurrence: one-year results of the APACC trial. AB - BACKGROUND & AIMS: Epidemiologic and experimental studies have suggested that aspirin intake reduces the risk for colorectal carcinogenesis. However, the available data are not sufficient to serve as the basis for firm recommendations. METHODS: We randomly assigned 272 patients with a history of colorectal adenomas (at least one more than 5 mm in diameter, or more than 3) to daily lysine acetylsalicylate (160 or 300 mg/day) or placebo for 4 years. The primary end points were adenoma recurrence after 1 and 4 years. These results are those of the year 1 colonoscopy. RESULTS: Among the 238 patients who completed the year 1 colonoscopy, at least one adenoma was observed in 38 patients of the 126 (30%) in the aspirin group and in 46 of the 112 (41%) in the placebo group; relative risk was 0.73 (95% confidence interval [CI]: 0.52-1.04; P = 0.08). At least one adenoma of more than 5 mm diameter was observed in 13 patients (10%) in the aspirin group and 26 (23%) in the placebo group (P = 0.01). The corresponding numbers for adenomas more than 10 mm in diameter were one (1%) and 7 (6%) (P = 0.05). Stepwise regression showed that independent factors associated with lower adenoma recurrence are aspirin treatment (adenoma >5 mm, P = 0.01), absence of personal history of adenoma before the entry colonoscopy (P = 0.01), and initial adenomatous polyp burden less than 10 mm (P = 0.001). CONCLUSIONS: Daily soluble aspirin is associated with a reduction in the risk for recurrent adenomas found at colonoscopy 1 year after starting treatment. PMID- 12891534 TI - Intestinal T-cell responses to high-molecular-weight glutenins in celiac disease. AB - BACKGROUND & AIMS: The chronic, small intestinal inflammation that defines celiac disease is initiated by a HLA-DQ2 restricted T-cell response to ingested gluten peptides after their in vivo deamidation by tissue transglutaminase (TG2). To date, celiac disease can only be treated by a lifelong abstinence from foods that contain wheat, rye, or barley; better therapeutic options are hence needed. An attractive target would be to identify nontoxic wheat cultivars or components thereof with intact baking qualities. Because these qualities are mainly determined by the high molecular weight (HMW) glutenin proteins of gluten, it is critical to know if these proteins are toxic or, more specifically, if they will trigger the activation of T cells in the celiac lesion. METHODS: Different, highly purified HMW glutenins were isolated from wheat cultivars or expressed as recombinant proteins. The proteins were first tested for recognition by a large panel of gluten-specific T-cell lines established from celiac lesions and then applied during ex vivo challenges of celiac biopsies to allow for a direct identification of HMW specific T cells. RESULTS: Intestinal T-cell responses to TG2-deamidated HMW glutenins but not the corresponding native proteins were detectable in 9 of the 22 adult and childhood celiac disease patients tested. CONCLUSIONS: T cells within celiac lesions frequently recognize deamidated HMW glutenin proteins. This finding questions the possibility of implementing these proteins in novel food items destined to be nontoxic for celiac disease patients. PMID- 12891535 TI - Human small-intestinal epithelium contains functional natural killer lymphocytes. AB - BACKGROUND & AIMS: CD3(-) non-T lymphocytes constitute the second most abundant lymphoid subset in the human small-bowel epithelium, and these CD3(-) intraepithelial lymphocytes are virtually absent in active celiac disease. Phenotypically, they resemble natural killer cells and have been termed natural killer-like intraepithelial lymphocytes. Because of the limited availability of appropriate human samples, functional studies have not yet been reported, and it is not yet clear whether these are true natural killer cells. METHODS: We used magnetic bead-based purification and flow cytometry to study several aspects of normal human small-bowel natural killer-like intraepithelial lymphocytes: intracellular cytokine content (basally and after activation); ability to lyse natural killer-sensitive K562 target cells; and expression of perforins, Fas ligand, and other functional markers. RESULTS: CD3(-) intraepithelial lymphocytes cultured in interleukin-2 showed a higher lymphokine-activated killer activity than CD3(+) intraepithelial lymphocytes (48%-83% lysis exerted by CD3(-) intraepithelial lymphocytes at an effector-target cell ratio of 2:1 vs. 8%-18% by CD3(+) intraepithelial lymphocytes). Perforin content correlated with this lytic potential (75% +/- 4% in CD3(-) vs. 5% +/- 4% in CD3(+) intraepithelial lymphocytes). Both CD3(-) and CD3(+) cells displayed a type I cytokine profile (interferon-gamma > tumor necrosis factor-alpha > interleukin-2; undetectable interleukin-4 and interleukin-10). In addition to their activated phenotype, subsets of natural killer-like intraepithelial lymphocytes expressed CD8alphaalpha and intracellular CD3epsilon chain, showing the existence of heterogeneity within this cell lineage. CONCLUSIONS: This is the first demonstration of functional natural killer cells within the human gut epithelium. These cells might play an important role in innate mucosal immunity (host defense and tumor surveillance) and tolerance. PMID- 12891536 TI - Childhood constipation: longitudinal follow-up beyond puberty. AB - BACKGROUND & AIMS: Sparse data exist about the prognosis of childhood constipation and its possible persistence into adulthood. METHODS: A total of 418 constipated patients older than 5 years at intake (279 boys; median age, 8.0 yr) participated in studies evaluating therapeutic modalities for constipation. All children subsequently were enrolled in this follow-up study with prospective data collection after an initial 6-week intensive treatment protocol, at 6 months, and thereafter annually, using a standardized questionnaire. RESULTS: Follow-up was obtained in more than 95% of the children. The median duration of the follow-up period was 5 years (range, 1-8 yr). The cumulative percentage of children who were treated successfully during follow-up was 60% at 1 year, increasing to 80% at 8 years. Successful treatment was more frequent in children without encopresis and in children with an age of onset of defecation difficulty older than 4 years. In the group of children treated successfully, 50% experienced at least one period of relapse. Relapses occurred more frequently in boys than in girls (relative risk 1.73; 95% confidence interval, 1.15-2.62). In the subset of children aged 16 years and older, constipation still was present in 30%. CONCLUSIONS: After intensive initial medical and behavioral treatment, 60% of all children referred to a tertiary medical center for chronic constipation were treated successfully at 1 year of follow-up. One third of the children followed up beyond puberty continued to have severe complaints of constipation. This finding contradicts the general belief that childhood constipation gradually disappears before or during puberty. PMID- 12891537 TI - A proinflammatory genetic profile increases the risk for chronic atrophic gastritis and gastric carcinoma. AB - BACKGROUND & AIMS: Pro-inflammatory polymorphisms within the genes interleukin (IL)-1B and IL-1RN are associated with risk for gastric carcinoma (GC) in Helicobacter pylori-infected individuals. We aimed to determine the association between variation of the tumor necrosis factor (TNF)-alpha gene and the risk for chronic atrophic gastritis (CAG) and GC. We also investigated the extent to which the combined effect of proinflammatory genetic polymorphisms (IL-1B, IL-1RN, and TNF-alpha), and the combined effect of TNF-alpha and bacterial genotypes each influence such a risk. METHODS: In a case-control study including 306 controls, 221 individuals with chronic gastritis, and 287 GC patients, the TNF-alpha-308 and IL-1B-511 bi-allelic polymorphisms, the IL-1RN variable number of tandem repeats (VNTR), and the H. pylori genes vacA (s and m regions) and cagA were genotyped. RESULTS: We found that carriers of the TNF-alpha-308*A allele are at increased risk for GC development with an odds ratio (OR) of 1.9 (95% confidence interval [CI], 1.3-2.7). For both CAG and GC, the odds of developing disease increased with the number of high-risk genotypes. Individuals carrying high-risk genotypes at the 3 loci are at increased risk for CAG and GC with an OR of 5.8 (95% CI, 1.1-31.0) and 9.7 (95% CI, 2.6-36.0), respectively. The risk for GC was not affected significantly by the combination of bacterial and TNF-alpha-308 genotypes. CONCLUSIONS: These findings show that a proinflammatory polymorphism in the TNF-alpha gene is associated with increased risk for GC, and that it is possible to define a specific genetic profile associated with highest risk for CAG and GC. PMID- 12891538 TI - Rates of dyspepsia one year after Helicobacter pylori screening and eradication in a Danish population. AB - BACKGROUND & AIMS: Helicobacter pylori (Hp) is strongly correlated with peptic ulcer and is a risk factor for gastric cancer. The aim of this study was to assess whether screening and eradication of Hp in a general population would reduce the prevalence of dyspepsia and the incidence of peptic ulcer and thus save health care resources and improve quality of life. METHODS: Twenty thousand individuals aged 40 to 65 years were randomized to screening and eradication for Hp or to the control group. Hp status was assessed by a whole blood Hp test, a positive result confirmed by a (13)C-urea breath test. Hp-positive individuals were offered Hp eradication therapy. The prevalence of dyspepsia and the quality of life were assessed through a mailed questionnaire. Information on the use of endoscopies and the use of prescription medication was obtained from registers. RESULTS: The response rate was 62.6%. The prevalence of Hp was 17.5%. The Hp eradication rate was 95%. In the intervention group, the prevalence of dyspepsia decreased from 24.3% at inclusion to 20.5% at 1-year follow-up. The reduction was similar in Hp-negative and Hp-positive persons. In the control group, dyspepsia increased from 20.3% to 21.5%. Gastroesophageal reflux symptoms improved slightly in Hp-eradicated participants. Except for a decreased consultation rate for dyspepsia, there were no visible savings in health care. CONCLUSIONS: Dyspepsia was modestly reduced after the screening and treatment procedure, and the result was not sufficiently extensive to have an effect on the use of health care or to improve quality of life. PMID- 12891539 TI - Tacrolimus for the treatment of fistulas in patients with Crohn's disease: a randomized, placebo-controlled trial. AB - BACKGROUND & AIMS: This study determined the effectiveness of tacrolimus for the treatment of Crohn's disease fistulas. METHODS: The study was a randomized, double-blind, placebo-controlled, multicenter clinical trial. Forty-eight patients with Crohn's disease and draining perianal or enterocutaneous fistulas were randomized to treatment with oral tacrolimus 0.2 mg. kg(-1). day(-1) or placebo for 10 weeks. The primary outcome measure was fistula improvement as defined by closure of >/=50% of particular fistulas that were draining at baseline and maintenance of that closure for at least 4 weeks. A secondary outcome measure was fistula remission as defined by closure of all fistulas and maintenance of that closure for at least 4 weeks. RESULTS: Forty-three percent of tacrolimus-treated patients had fistula improvement compared with 8% of placebo treated patients (P = 0.004). Ten percent of tacrolimus-treated patients had fistula remission compared with 8% of placebo-treated patients (P = 0.86). Adverse events significantly associated with tacrolimus, including headache, increased serum creatinine level, insomnia, leg cramps, paresthesias, and tremor, were managed with dose reduction. CONCLUSIONS: Oral tacrolimus 0.2 mg. kg(-1). day(-1) is effective for fistula improvement, but not fistula remission, in patients with perianal Crohn's disease. Adverse events associated with tacrolimus can be managed by dose reduction. Lower doses of tacrolimus should be evaluated. PMID- 12891540 TI - Gastrointestinal health care resource utilization with chronic use of COX-2 specific inhibitors versus traditional NSAIDs. AB - BACKGROUND & AIMS: Cyclooxygenase 2 (COX-2)-specific inhibitors (coxibs) decrease gastrointestinal (GI) events in controlled trials, but results in clinical practice are unknown. We assessed GI-related resource use and costs in patients switching from chronic nonsteroidal anti-inflammatory drug (NSAID) therapy to chronic coxib therapy and in patients starting chronic NSAID therapy vs. chronic coxib therapy in a U.S. administrative claims database of >8 million lives. METHODS: "Switchers" (n = 2246) were assessed in the 12-month periods before and after switching from chronic NSAID therapy to coxib therapy. "New NSAID" (n = 25,989) and "new coxib" (n = 2125) groups were assessed for the 12-month periods before and after the initial prescription. Proportions of patients with GI resource use (odds ratio [OR] adjusted for relevant covariates) and costs were compared. RESULTS: The adjusted OR for any GI resource use (coxib vs. NSAID period) among switchers was 0.86 (0.74-0.99). The decrease was due to less GI cotherapy (OR = 0.82 [0.69-0.97]). Costs were not significantly lower after switching to coxibs (mean difference, -$19; 95% CI: -$139, $55), although after adding NSAID/coxib costs, the total cost in the coxib period was significantly higher (mean increase, $377; $271, $488). Adjusted OR for GI resource use for new coxib vs. new-NSAID was 1.04 (0.92-1.16), but GI costs were significantly lower in new-NSAID patients. CONCLUSIONS: Patients switching from chronic NSAID therapy to chronic coxib therapy had a slight decrease in the proportion using GI-related resources but not in GI costs. When NSAID/coxib drug costs were included, costs were significantly less with NSAIDs than with coxibs. The potential GI-related cost savings suggested in coxib clinical trials may not be fully realized in "real-world" settings. PMID- 12891541 TI - A randomized trial comparing heater probe plus thrombin with heater probe plus placebo for bleeding peptic ulcer. AB - BACKGROUND & AIMS: This multicenter, double-blind, controlled trial compared the efficacy of combined endoscopic hemostatic treatment using the heater probe plus thrombin injection with that of the heater probe plus placebo injection as treatment for peptic ulcers with active bleeding or nonbleeding visible vessels. Efficacy was defined in terms of primary hemostasis, prevention of rebleeding, and need for urgent surgery. METHODS: Two hundred forty-seven patients presenting with major peptic ulcer bleeding were randomized to heater probe plus thrombin or to heater probe plus placebo. The groups were well matched for all risk categories including age, endoscopic stigmata, shock, and severity of comorbid diseases. Endoscopic therapy was applied using the heater probe followed by injection of thrombin or placebo. RESULTS: Successful primary hemostasis was achieved in 97% of patients. Rebleeding developed in 19 (15%) of thrombin plus heater probe patients and 17 (15%) of placebo plus heater probe patients. Emergency surgery was necessary in 16 and 13 patients, respectively. Eight patients in the thrombin group had adverse events compared with 4 in the placebo group. Eight (6%) of thrombin plus heater probe patients and 14 (12%) of placebo plus heater probe patients died (P = 0.21). CONCLUSIONS: The combination of thrombin and the heater probe does not confer an additional benefit over heater probe and placebo as endoscopic treatment for bleeding peptic ulcer. Our trial does not support the use of this combination of hemostatic therapy. PMID- 12891542 TI - The cyclooxygenase-2-selective inhibitors rofecoxib and celecoxib prevent colorectal neoplasia occurrence and recurrence. AB - BACKGROUND & AIMS: Colorectal cancer is one of the leading causes of cancer death. Most colorectal cancers are believed to develop from colorectal adenomas. We examined the effect of the selective cyclooxygenase-2 inhibitors rofecoxib and celecoxib, nonselective nonsteroidal anti-inflammatory drugs, aspirin, and acetaminophen on colorectal neoplasia (colorectal cancer, colorectal adenoma, or both). METHODS: This was a nested case-control study, which used data from a government insurance database on patients 65 years and older who underwent a diagnostic test or procedure for colorectal neoplasia between January and June 2001. Logistic regression models were used to determine the effect of exposure to the drugs of interest for at least 3 months on the occurrence or recurrence of colorectal neoplasia. RESULTS: The control group included 2568 patients found to be free of colorectal neoplasia; 730 patients were diagnosed with colorectal adenoma, and 179 were diagnosed with colorectal cancer. Patients more likely to have colorectal adenoma (odds ratio, 95% confidence interval) were those diagnosed with colorectal adenoma (4.12, 3.27-5.18) or colorectal cancer (3.74, 2.32-6.03) in the previous 1-3 years and those with hemorrhage of the rectum or unspecified anemia in the prior month (3.19, 2.46-4.12). Exposures to rofecoxib (0.67, 0.46-0.98) and nonselective nonsteroidal anti-inflammatory drugs (0.41, 0.21-0.83) reduced the risk of colorectal adenoma. Rofecoxib, celecoxib, and nonselective nonsteroidal anti-inflammatory drugs were all protective against both neoplasias (0.64, 0.45-0.91; 0.73, 0.54-0.99; and 0.47, 0.26-0.86, respectively). CONCLUSIONS: Rofecoxib, celecoxib, and nonselective nonsteroidal anti-inflammatory drugs seem to protect against the development of colorectal neoplasia. PMID- 12891543 TI - Clostridium difficile toxin B is an inflammatory enterotoxin in human intestine. AB - BACKGROUND & AIMS: Clostridium difficile causes antibiotic-associated diarrhea and pseudomembranous colitis, diseases afflicting millions of people each year. Although C. difficile releases 2 structurally similar exotoxins, toxin A and toxin B, animal experiments suggest that only toxin A mediates diarrhea and enterocolitis. However, toxin A-negative/toxin B-positive strains of C. difficile recently were isolated from patients with antibiotic-associated diarrhea and colitis, indicating that toxin B also may be pathogenic in humans. METHODS: Here we used subcutaneously transplanted human intestinal xenografts in immunodeficient mice to generate a chimeric animal model for C. difficile toxin induced pathology of human intestine. RESULTS: We found that intraluminal toxin B, like equivalent concentrations of toxin A, induced intestinal epithelial cell damage, increased mucosal permeability, stimulated interleukin (IL)-8 synthesis, and caused an acute inflammatory response characterized by neutrophil recruitment and tissue damage. Laser capture microdissection and real-time quantitative reverse-transcription polymerase chain reaction (RT-PCR) showed that intestinal epithelial cell-specific IL-8 gene expression also was increased significantly after luminal exposure to C. difficile toxins in vivo. CONCLUSIONS: We conclude that C. difficile toxin B, like toxin A, is a potent inflammatory enterotoxin for human intestine. Future therapeutic or vaccine strategies for C. difficile infection therefore need to target both toxins. PMID- 12891544 TI - Gastric electrical stimulation for medically refractory gastroparesis. AB - BACKGROUND & AIMS: This study investigated the efficacy of gastric electrical stimulation for the treatment of symptomatic gastroparesis unresponsive to standard medical therapy. METHODS: Thirty-three patients with chronic gastroparesis (17 diabetic and 16 idiopathic) received continuous high frequency/low-energy gastric electrical stimulation via electrodes in the muscle wall of the antrum connected to a neurostimulator in an abdominal wall pocket. After implantation, patients were randomized in a double-blind crossover design to stimulation ON or OFF for 1-month periods. The blind was then broken, and all patients were programmed to stimulation ON and evaluated at 6 and 12 months. Outcome measures were vomiting frequency, preference for ON or OFF, upper gastrointestinal tract symptoms, quality of life, gastric emptying, and adverse events. RESULTS: In the double-blind portion of the study, self-reported vomiting frequency was significantly reduced in the ON vs. OFF period (P < 0.05) and this symptomatic improvement was consistent with the significant patient preference (P < 0.05) for the ON vs. OFF period determined before breaking the blind. In the unblinded portion of the study, vomiting frequency decreased significantly (P < 0.05) at 6 and 12 months. Scores for symptom severity and quality of life significantly improved (P < 0.05) at 6 and 12 months, whereas gastric emptying was only modestly accelerated. Five patients had their gastric electrical stimulation system explanted or revised because of infection or other complications. CONCLUSIONS: High-frequency/low-energy gastric electrical stimulation significantly decreased vomiting frequency and gastrointestinal symptoms and improved quality of life in patients with severe gastroparesis. PMID- 12891545 TI - Fracture risk in people with celiac disease: a population-based cohort study. AB - BACKGROUND & AIMS: People with celiac disease are at risk of developing osteoporosis, but the extent of any increased fracture risk is unclear. We performed a population-based cohort study by using the General Practice Research Database to quantify the fracture risk in people with celiac disease compared with the general population. METHODS: We identified 4732 people with celiac disease, of whom 1589 were "incident" cases, and 23,620 age- and sex-matched control subjects. We used Cox regression to estimate the hazard ratios for any fracture, hip fracture, and ulna or radius fracture in the celiac disease cohort compared with the general population. RESULTS: In the incident subjects with celiac disease, the mean age at diagnosis was 44 years, and 67% were women. The overall hazard ratio for any fracture was 1.30 (95% confidence interval, 1.16 1.46), for hip fracture was 1.90 (95% confidence interval, 1.20-3.02), and for ulna or radius fracture was 1.77 (95% confidence interval, 1.35-2.34). The absolute difference in the overall fracture rate was 3.20 per 1000 person-years and for hip fracture it was 0.97 per 1000 person-years in those older than 45 years. In 1589 incident subjects, the excess fracture risk was slightly lower compared with the "prevalent" subjects (hazard ratio for any fracture, 1.19 vs. 1.40, respectively). CONCLUSIONS: There were small increases in both the absolute and relative fracture risks in people with celiac disease; the excess risks were slightly lower in those with a more recent diagnosis. Our data indicate that concerns regarding a markedly increased fracture risk in celiac disease are unwarranted. PMID- 12891546 TI - Hepatocyte apoptosis and fas expression are prominent features of human nonalcoholic steatohepatitis. AB - BACKGROUND & AIMS: The pathogenesis of nonalcoholic steatohepatitis (NASH) remains poorly understood. Although apoptosis is a common mechanism of liver injury, the extent and clinical significance of apoptosis in NASH has not been examined. Thus, the aims of this study were to quantify hepatocyte apoptosis in NASH, correlate it with disease severity, and identify possible mechanisms of apoptosis induction. METHODS: Hepatocyte apoptosis was assessed in NASH, simple steatosis, alcoholic hepatitis, and controls without liver disease using the TUNEL assay and immunohistochemistry for activated caspases 3 and 7. Liver specimens were also graded according to the magnitude of inflammation and fibrosis. RESULTS: TUNEL-positive cells were significantly increased in liver biopsy specimens from patients with NASH compared with simple steatosis and controls. Unexpectedly, TUNEL-positive cells were also greater in NASH vs. alcoholic hepatitis. Immunohistochemistry demonstrated active caspases 3 and 7 in NASH specimens, confirming the occurrence of apoptosis in this disease. A positive correlation was observed between hepatocyte apoptosis and hepatic fibrosis and inflammatory activity, respectively. The Fas receptor was strongly expressed in hepatocytes in liver specimens from NASH patients as compared with controls. CONCLUSIONS: Hepatocyte apoptosis is significantly increased in patients with NASH and correlates with disease severity, suggesting that antiapoptotic therapy may be useful in this syndrome. PMID- 12891547 TI - Hepatitis B virus genotypes in the United States: results of a nationwide study. AB - BACKGROUND & AIMS: Hepatitis B virus (HBV) genotypes may be related to severity of liver disease and treatment response. The aims of this nationwide study were to determine the prevalence of HBV genotypes in the United States and the association between HBV genotypes and patient demographics, mode of infection, and clinical status. METHODS: A total of 694 consecutive chronic HBV-infected patients seen in 17 U.S. liver centers during a 1-year period were enrolled. Demographic, clinical, and laboratory data were collected. Sera were tested for HBV genotyping, precore, and core promoter variants by line-probe assays. RESULTS: All 7 HBV genotypes (A-G) were found, with genotypes A and C the most common. The prevalence of HBV genotypes was different in different regions of the United States. A strong correlation was found between HBV genotypes and ethnicity. HBV genotype A was prevalent among white and black patients, whereas genotypes B and C were most common among Asian patients. The predominant genotype among patients born in the United States, Europe, the Far East, and Southeast Asia were A, D, C, and B, respectively. Genotypes A and C were associated with a higher prevalence of hepatitis B e antigen. Precore variant was detected in 27% of patients and core promoter variant in 44% of patients. CONCLUSIONS: Our study suggests that the epidemiology of HBV infection in the United States may have changed over time as a result of immigration from countries with a high prevalence of HBV infection. HBV genotypes may account for the heterogeneity in disease manifestations among patients with chronic HBV infection. PMID- 12891548 TI - ABCB4 gene mutation-associated cholelithiasis in adults. AB - BACKGROUND & AIMS: We recently put forward arguments in favor of ABCB4 gene (adenosine triphosphate-binding cassette, subfamily B, member 4) defects as a risk factor for symptomatic cholelithiasis in adults. In this study, we characterized ABCB4 gene mutations in a series of patients with symptomatic cholelithiasis to determine the genetic basis and the clinical phenotype of ABCB4 gene mutation-associated cholelithiasis. METHODS: We analyzed the entire ABCB4 gene coding sequences in a first group of 32 patients who had a clinical history compatible with the syndrome previously described, in a second group of 28 patients who presented with a classic gallstone disease that justified a cholecystectomy, and in a third group of 33 patients without a history of cholelithiasis. RESULTS: We identified both heterozygous and homozygous ABCB4 gene point mutations in 18 of 32 (56%) patients who presented with clinical criteria of the syndrome, whereas no mutation was detected in the 2 other groups of patients (P < 0.001). Three independent clinical features were strongly associated with point mutations: recurrence of symptoms after cholecystectomy (odds ratio, 8.5); intrahepatic hyperechoic foci, intrahepatic sludge, or microlithiasis (odds ratio, 6.1); and age <40 years at the onset of symptoms (odds ratio, 3.0). ABCB4 gene point mutations were detected exclusively in the patients who showed 2 or 3 of these clinical features. CONCLUSIONS: Our results show that ABCB4 gene mutations represent a major genetic risk factor in a symptomatic and recurring form of cholelithiasis in young adults. PMID- 12891549 TI - Heme oxygenase-1 is an antifibrogenic protein in human hepatic myofibroblasts. AB - BACKGROUND & AIMS: Hepatic myofibroblasts play a key role in the development of liver fibrosis associated with chronic liver diseases. We have shown that oxidative stress is a messenger of 15-deoxy-delta-12,14-prostaglandin J2 (15-d PGJ2) in human hepatic myofibroblasts. The aim of the present study was to investigate the role of a stress-inducible protein, heme oxygenase-1 (HO-1), in the action of 15-d-PGJ2. METHODS: Expression of HO-1 was characterized in biopsy specimens of normal human liver and active cirrhosis by immunohistochemistry, and in cultured human hepatic myofibroblasts by Northern and Western blot analysis. Functional studies also were performed in cultured human hepatic myofibroblasts. RESULTS: Immunohistochemistry showed that in biopsy specimens from normal livers, HO-1 protein expression was restricted to Kupffer cells. Biopsy specimens from cirrhotic patients displayed HO-1 protein both in macrophages and in myofibroblasts within fibrotic septa. HO-1 messenger RNA (mRNA) and protein also were detected in cultured human hepatic myofibroblasts and increased in response to 15-d-PGJ2 in a time- and dose-dependent manner. Induction of HO-1 in human hepatic myofibroblasts mediated 2 major antifibrogenic properties of 15-d-PGJ2, namely, inhibition of proliferation and of procollagen I mRNA expression. These effects were ascribed to bilirubin, one of the products of HO-1-mediated heme degradation. CONCLUSIONS: This study shows that HO-1 is expressed in human hepatic myofibroblasts and induced during chronic liver injury. Moreover, these data unravel HO-1 as a major antifibrogenic pathway. PMID- 12891550 TI - Diarrhea caused by enterotoxigenic Escherichia coli infection of humans is inhibited by dietary calcium. AB - BACKGROUND & AIMS: In several rat infection experiments, we have shown that dietary calcium inhibits intestinal colonization and translocation of invasive salmonella. The aim of the present study was to find out whether calcium is also protective against enterotoxigenic Escherichia coli (ETEC) infection. This was first tested in our rat model and subsequently verified in a human infection study. METHODS: Rats were fed a purified diet with either a low or a high amount of calcium phosphate and orally infected with ETEC. In addition, a parallel, double-blind, placebo-controlled intervention study of 3 weeks was performed with 32 healthy men. Subjects largely maintained their habitual diet and consumed either regular milk products (calcium supply, 1100 mg/day) or placebo milk products (calcium supply, 60 mg/day). On day 10, subjects ingested a live but attenuated ETEC strain (strain E1392/75-2A), able to induce mild although short lived symptoms. Primary outcomes studied were infection-induced diarrhea (total fecal output and relative fecal dry weight) and fecal mucin excretion. RESULTS: In humans, ETEC induced diarrhea in both groups, in that total fecal output doubled and mean relative fecal dry weight dropped from 25% to 20%. Additionally, fecal mucin excretion was increased in both groups. All these fecal parameters were completely normalized in the calcium group on the second infection day, in contrast to the placebo group, which recovered on the third infection day. Likewise, supplemental calcium inhibited ETEC colonization and diarrhea in rats. CONCLUSIONS: Calcium in milk products improves human resistance to ETEC infection as it inhibits infectious diarrhea. PMID- 12891551 TI - Identification of a quantitative trait locus for ileitis in a spontaneous mouse model of Crohn's disease: SAMP1/YitFc. AB - BACKGROUND & AIMS: The SAMP1/Yit mouse strain develops spontaneous ileitis with histologic features of Crohn's disease. Disease expression in the SAMP1/YitFc subline (SAMP1/Fc) is partially inhibited by outcross to C57BL/6J (B6) mice, suggesting complex genetic control of disease susceptibility with both dominant and recessive determinants. We performed a genetic analysis of a (B6 x SAMP1/Fc)F(2) cross to localize the genes regulating intestinal inflammation in this model. METHODS: A genome-wide scan was performed using a panel of microsatellite loci determined to be informative for this cross. Quantitative trait loci were identified with Map Manager QT using a serial regression approach. Positional candidate genes were selectively sequenced at the genomic level to identify potential susceptibility genes for functional screening. RESULTS: A genome-wide scan of (B6 x SAMP1/Fc)F(2) mice identified a SAMP-derived quantitative trait loci with additive effects on chromosome 9 in a region likely to have been inherited from the AKR mouse strain. The candidate interval contains several genes of interest because of their potential role in either immune system function, intestinal epithelial function, or both. Suggestive evidence for additional loci was also observed on chromosomes 6 and X. CONCLUSIONS: The SAMP1/Fc allele for a locus, designated Ibdq1, promotes inflammation-associated epithelial damage in these mice. Consistent with persistent mild ileitis in (B6 x SAMP1/Fc)F(1) mice, this locus appears to function in an additive fashion. Two genes in this interval, encoding the interleukin 10 receptor alpha chain and interleukin 18, are excellent candidates for Ibdq1. PMID- 12891552 TI - CCR5 mediates specific migration of Toxoplasma gondii-primed CD8 lymphocytes to inflammatory intestinal epithelial cells. AB - BACKGROUND & AIMS: Toxoplasma gondii, an obligate intracellular parasite, can invade intestinal epithelial cells and elicit a robust Th1 immune response. In this model of intestinal inflammation, CD8(+) intraepithelial lymphocytes (IELs) secrete transforming growth factor (TGF)-beta, which appears necessary for the maintenance of homeostasis in the intestine. However, the mechanism responsible for the IEL migration to the inflamed intestine is still unclear. METHODS: An in vitro coculture cell system was used to quantify the IEL attraction by an infected intestinal epithelial cell line (m-IC(cl2)). We used CCR5-deficient mice to determine which chemokine receptor-chemokine interaction could be responsible for the recruitment of antigen-specific CD8(+) IELs to the small intestine for the promotion of parasite clearance and host recovery. RESULTS: We observed increased expression of several chemokine receptors (CCR1, CCR2, CCR5, CXCR3) in the infected ileum. In particular, CCR5 expression was markedly increased in antigen-primed CD8(+) IELs. Experiments using recombinant chemokines as well as blocking antibodies showed that macrophage inflammatory protein (MIP)-1alpha and MIP-1beta were critical for their homing. CD8(+) IELs isolated from CCR5 deficient mice (CCR5-/-), despite their high production of TGF-beta and overexpression of activation markers, were impaired in their ability to migrate in vitro to the m-IC(cl2) monolayer or in vivo to the inflamed intestine after adoptive transfer. CONCLUSIONS: Our data emphasize the biologic role of CCR5 as an important component in the migration of intraepithelial CD8(+) T cells and the regulation of the inflammatory response following parasite infection. PMID- 12891553 TI - Pathogenicity of the hereditary colorectal cancer mutation hMLH1 del616 linked to shortage of the functional protein. AB - BACKGROUND & AIMS: Hereditary nonpolyposis colorectal cancer is associated with mismatch repair deficiency. Most predisposing mutations prevent the production of functional mismatch repair protein. Thus, when the wild-type copy is also inactivated, the cell becomes mismatch repair deficient, and this leads to a high degree of microsatellite instability in tumors. However, tumors linked to nontruncating mutations may display positive or partly positive immunohistochemical staining of the mutated protein and low or atypical microsatellite instability status, which suggests impaired functional activity but not a total lack of mismatch repair. We found human mutL homology (hMLH) 1 del616, one of the most widespread recurring mutations in hereditary nonpolyposis colorectal cancer, segregating in a large hereditary nonpolyposis colorectal cancer family. Because the predicted coding change is a deletion of only 1 amino acid, the pathogenicity of the mutation was evaluated. METHODS: Many analyses were performed to assess the pathogenicity of hMLH1 del616 and to study the expression and function of the mutated messenger RNA and protein. RESULTS: Genetic and immunohistochemical evidence supported hMLH1-linked cancer predisposition in this family. Microsatellite instability varied from low to high, and the hMLH1 protein was lost in 2 tumors but was partly detectable in 1 tumor. Whereas similar optimal amounts of mutated hMLH1 del616 and wild-type hMLH1 proteins were equally functional in an in vitro mismatch repair assay, the amount of in vivo-expressed hMLH1 del616 was much lower than the amount of wild type protein; this suggests that the deletion imparts instability to the mutant protein. CONCLUSIONS: Our results suggest that the pathogenicity of hMLH1 del616 is not linked to nonfunctionality, but to shortage of the functional protein. PMID- 12891554 TI - Transcriptional regulation of the human trefoil factor, TFF1, by gastrin. AB - BACKGROUND & AIMS: This study aimed to identify gastrin-sensitive genes that may mediate the effects of this hormone on gastric epithelial architecture. METHODS: Gastrin-sensitive genes were identified by messenger RNA (mRNA) differential display of the gastric fundus from gastrin-deficient (GAS-KO) or wild-type mice. Gastrin-stimulated expression of the trefoil peptide TFF1 in mouse fundus and in the gastric cancer cell line AGS-G(R) was determined by Northern blot and real time polymerase chain reaction. Transcriptional regulation of TFF1 in AGS-G(R) cells was studied using promoter-reporter assays and electrophoretic mobility shift assay. Expression of TFF1 and the cholecystokinin(B) receptor in response to gastric mucosal injury was determined by immunohistochemistry. RESULTS: mRNA differential display identified TFF1 as a gastrin-regulated gene. TFF1 mRNA was reversibly reduced in GAS-KO mice and increased in a hypergastrinemic transgenic strain versus respective background strains. TFF1 mRNA expression was rapidly and potently induced by gastrin in a gastric cancer cell line that expresses the gastrin/cholecystokinin(B) receptor. Gastrin responsiveness of the human TFF1 promoter mapped to a G-C rich region 300 base pairs upstream of the transcriptional start site. This region bound the transcription factors SP3 and MAZ. Gastrin activated transcription through a Raf-, Mek- and Erk-dependent but Ras-independent pathway. TFF1 expression was induced both directly and by transactivation between neighboring cells. Neither direct nor indirect gastrin induced TFF1 expression required activation of the epidermal growth factor receptor. CONCLUSIONS: Gastrin exerts tonic control of TFF1 expression but also has the potential for rapid up-regulation of this trefoil factor. TFF1 is a potential candidate to counterbalance the proliferative effects of gastrin. PMID- 12891555 TI - Transduction of the liver with activated Akt normalizes portal pressure in cirrhotic rats. AB - BACKGROUND & AIMS: Portal hypertension in cirrhosis is secondary to an increase in hepatic resistance that occurs mainly through collagen deposition. However, recent evidence points to a major contribution by other factors, such as an intrahepatic reduction in nitric oxide production. Akt is a major activator of the endothelial nitric oxide synthase (eNOS) enzyme, but its potential role in intrahepatic resistance in cirrhosis is unknown. For this reason the aims of the present study were to determine whether there is an impaired Akt activation in cirrhotic livers and how this phenomenon relates to the decrease in NO production associated with portal hypertension. METHODS: Cirrhosis was induced in rats by carbon tetrachloride inhalation. Protein abundance and phosphorylation status of Akt and eNOS were examined by Western blotting. The role of Akt in the liver of cirrhotic rats was investigated through infection with adenoviruses encoding either beta-galactosidase (beta-gal) or constitutively active Akt (myr-Akt). RESULTS: The liver of cirrhotic animals showed a significant reduction in Akt and eNOS phosphorylation. Adenoviral delivery of myr-Akt restored eNOS phosphorylation and increased the intrahepatic concentration of guanosine 3',5' cyclic monophosphate. These events were associated with normalization in portal pressure and a significant increase in mean arterial pressure after 3 days of adenoviral infection. In contrast, transduction of livers with beta-gal did not produce any change in these hemodynamic parameters. CONCLUSIONS: myr-Akt gene therapy restored Akt activation and NO production in the cirrhotic liver, suggesting that this therapy may be useful for the treatment of portal hypertension. PMID- 12891556 TI - Lack of gp130 expression in hepatocytes promotes liver injury. AB - BACKGROUND & AIMS: Interleukin 6 (IL-6) contributes via its signal transducer gp130 to the acute phase response (APR) in hepatocytes. Recent studies indicated that IL-6 is involved in the regulation of different pathophysiologic conditions of the liver. To define the IL-6-dependent intracellular pathways more specifically, we generated a hepatocyte-specific gp130 knockout mouse. METHODS: Hepatocyte-specific gp130-deficient mice were generated using the Cre-loxP system. Expression of the Cre recombinase was under the control of a hepatocyte specific control element. Adult mice were challenged with IL-6, oncostatin M (OSM), and LPS. RESULTS: Cre expression started at day 10.5 postconception, and a complete deletion of gp130 in hepatocytes was found at day 14 during liver development. The adult liver of these mice showed no abnormalities; however, after IL-6 and OSM stimulation, gp130-dependent pathways (STAT3, APR gene expression) were completely blocked in the liver of these animals. Additionally, challenging hepatocyte-specific gp130 knockout animals with lipopolysaccharides (LPS) lead to an onset of acute liver injury with an increase of hepatocyte apoptosis associated with elevated tumor necrosis factor alpha (TNF-alpha) serum levels and reduced nuclear factor kappaB (NF-kappaB) activation in hepatocytes. CONCLUSIONS: Our findings demonstrate that gp130 is of minor relevance for embryonal development of hepatocytes. However, the molecule has an essential role in controlling acute phase gene expression and provides hepatocellular protection after LPS challenge. PMID- 12891557 TI - Farnesoid X receptor agonists suppress hepatic apolipoprotein CIII expression. AB - BACKGROUND & AIMS: Increased serum triglyceride levels constitute a risk factor for coronary heart disease. Apolipoprotein CIII (Apo CIII) is a determinant of serum triglyceride metabolism. In this study, we investigated whether activators of the nuclear farnesoid X receptor (FXR) modulate Apo CIII gene expression. METHODS: The influence of bile acids and synthetic FXR activators on Apo CIII and triglyceride metabolism was studied in vivo by using FXR wild-type and FXR deficient mice and in vitro by using human primary hepatocytes and HepG2 cells. RESULTS: In mice, treatment with the FXR agonist taurocholic acid strongly decreased serum triglyceride levels, an effect associated with reduced Apo CIII serum and liver messenger RNA levels. By contrast, no change was observed in FXR deficient mice. Incubation of human primary hepatocytes and HepG2 cells with bile acids or the nonsteroidal synthetic FXR agonist GW4064 resulted in a dose dependent down-regulation of Apo CIII gene expression. Promoter transfection experiments and mutation analysis showed that bile acid-activated FXR decrease human Apo CIII promoter activity via a negative FXR response element located in the I(4) footprint between nucleotides -739 and -704. Chromatin immunoprecipitation experiments showed that bile acid treatment led to binding of FXR/retinoid X receptor heterodimers to and displacement of HNF4alpha from this site. Bile acid treatment still repressed liver Apo CIII gene expression in hepatic HNF4alpha-deficient mice, suggesting an active rather than a competitive mechanism of Apo CIII repression by the FXR. CONCLUSIONS: We identified bile acid and synthetic activators of the nuclear FXR as negative regulators of Apo CIII expression, an effect that may contribute to the triglyceride-decreasing action of FXR agonists. PMID- 12891558 TI - JTE-522, a cyclooxygenase-2 inhibitor, is an effective chemopreventive agent against rat experimental liver fibrosis1. AB - BACKGROUND & AIMS: The aim of this study was to assess the effects of cyclooxygenase (COX)-2 inhibition on rat experimental liver fibrogenesis. METHODS: We investigated the inhibitory effects of a selective COX-2 inhibitor, JTE-522, on liver fibrosis induced by a choline-deficient, l-amino acid-defined diet (CDAA). Inhibitory effect was also tested in a second model of thioacetamide (TAA)-induced liver fibrosis. RESULTS: CDAA induced liver fibrosis and preneoplastic foci at 12 weeks and cirrhosis at 36 weeks. Hepatocellular carcinoma was noted in 13 of 15 rats (87%). JTE-522 significantly inhibited fibrosis and development of preneoplastic lesions in a dose-dependent manner and completely inhibited generation of cirrhosis and hepatocellular carcinoma at both low and high doses (10 and 30 mg/kg body wt/day, respectively). JTE-522 administrated only from 12 weeks to 36 weeks also prevented cirrhosis and formation of hepatocellular carcinoma. JTE-522 itself did not cause local or systemic gross or histopathologic changes at 36 weeks. Mechanistic studies indicated that the CDAA model displayed up-regulation of several biomarkers, including COX-2, arachidonate metabolite (prostaglandin E(2)), serum aspartate aminotransferase, and c-myc expression. The model also showed an increased proportion of activated hepatic stellate cells, proliferating cell nuclear antigen index, and CD45-positive inflammatory cells in the liver. JTE-522 effectively diminished these changes. JTE-522 exhibited similar antifibrosis effects in the TAA model. CONCLUSIONS: Our results suggest that COX-2 is involved in CDAA- and TAA-induced liver fibrosis. Our data also indicate that JTE-522 is a potent chemopreventive agent of rat liver fibrosis with low toxicity. PMID- 12891559 TI - Insulin signaling through insulin receptor substrate 1 and 2 in normal liver development. AB - BACKGROUND & AIMS: The insulin growth factor signal transduction pathway is an important regulator of adult hepatocyte proliferation. The purpose of this study was to determine the roles of the insulin receptor substrate (IRS-1 and IRS-2) mediated growth cascades in rapidly growing fetal rat liver. METHODS: We determined the expression and tyrosyl phosphorylation of the insulin receptor beta subunit (IRbeta), IRS-1 and IRS-2, the binding of phosphatidylinositol 3 kinase (PI3K), and activation of the mitogen-activated protein kinase (MAPK) pathway in the presence or absence of insulin stimulation in vivo during development and in the adult liver. In addition, activation of other downstream components including PI3K, Akt, GSK3beta, Bad, and p70S6 kinase was studied. RESULTS: We observed reduced expression and tyrosyl phosphorylation of IRS-1 in the fetal liver compared with the adult liver. These developmental changes resulted in a lack of sensitivity to insulin stimulation and subsequent downstream activation of the PI3K and MAPK cascades until the postneonatal period. In contrast, there was a high level of IRS-2 expression and insulin stimulated tyrosyl phosphorylation as early as embryonic day 15 with robust PI3K binding and activation, which may enhance hepatocyte survival during the rapid growth phase of the liver. CONCLUSIONS: The IRS-1 signal transduction pathway does not play a major role in fetal liver growth because IRS-2 functions as the major insulin responsive molecule in early development. However, insulin-mediated IRS-1/MAPK cascade activation contributes to growth in the adult. PMID- 12891560 TI - Gastric intestinal metaplasia on routine endoscopic biopsy. PMID- 12891561 TI - Pain as a complication of use of opiate antagonists for symptom control in cholestasis. AB - Controlled trials have suggested that opiate antagonist therapy may be effective for the treatment of the symptoms of cholestasis. The oral opiate antagonist naltrexone in particular has started to enter into routine clinical use for amelioration of cholestatic itch. Attention regarding the side effects of opiate antagonist therapy has, to date, largely focused on an opiate withdrawal-type reaction (which can be controlled effectively by titrated therapy introduction regimens). Here we describe 3 cases of a further clinically important side effect, loss of control of pain resulting from other pathologies, which in each case necessitated the withdrawal of hitherto clinically effective opiate antagonist therapy. Of the 14 patients treated by our unit with opiate antagonist agents for the control of cholestatic symptoms, 13 (93%) showed resolution of, or significant improvement in, symptoms. Of the 13 patients showing a clinical response, 7 (54%) subsequently had to discontinue therapy because of side effects (including the 3 patients with uncontrolled pain). It is our experience that in the routine clinical setting, opiate antagonists are highly effective for the treatment of cholestatic symptoms. In practice, however, their usefulness is limited by their side-effect profile. PMID- 12891564 TI - Is virtual colonoscopy ready for widespread application? PMID- 12891562 TI - Neutralizing antibodies in hepatitis C virus infection: a review of immunological and clinical characteristics. PMID- 12891565 TI - 6-mercaptopurine beats a bum rap. PMID- 12891566 TI - Will an aspirin a day keep the endoscope away? PMID- 12891567 TI - Iron and HFE mutations in nonalcoholic steatohepatitis: innocent bystanders or accessories to the crime? PMID- 12891568 TI - Cholecystectomy after treatment of choledocholithiasis: on firmer ground. PMID- 12891569 TI - The role of PPARgamma in colon cancer: is it as simple as APC? PMID- 12891570 TI - Combo cap takes a big bite, yet a smoothie to swallow. PMID- 12891571 TI - Portopulmonary hypertension: an increasingly important complication of cirrhosis. PMID- 12891576 TI - Discussion on the effect of chronic hypergastrinemia on human enterochromaffin like cells: insights from patients with sporadic gastrinomas. PMID- 12891577 TI - The prognosis for life in an HCV hyperendemic area. PMID- 12891578 TI - Reversal of cirrhosis: evidence-based medicine? PMID- 12891580 TI - Successful closure of ileal pouch-vaginal fistulas with infliximab. PMID- 12891581 TI - Differential p38 mitogen-activated protein kinase target phosphorylation in responders and nonresponders to infliximab. PMID- 12891584 TI - Hypnotherapy for functional dyspepsia: do the results add up? PMID- 12891585 TI - Selective cyclooxygenase-2 inhibitors and relapse of inflammatory bowel disease. PMID- 12891587 TI - The QUADRATE study: a proposal for a change in the reporting of pharmaceutical supported trials. PMID- 12891599 TI - Does gender affect the clinical outcome of patients with acute myocardial infarction complicated by cardiogenic shock who undergo percutaneous coronary intervention? AB - This study sought to determine the impact of female gender on clinical outcome in patients with acute myocardial infarction (AMI) complicated by cardiogenic shock (CS) due to predominant ventricular failure undergoing percutaneous coronary intervention (PCI). We analyzed gender-related differences in procedural, angiographic, and clinical outcomes in 208 consecutive patients with AMI complicated by CS. Out of 208 patients with CS, 65 were women and 143 men. Women were older than men (74 +/- 10 years vs. 66 +/- 12 years; P < 0.001) and had a greater incidence of a history of hypertension (43% vs. 29%; P = 0.041). The 6 month mortality rate was 42% in women and 31% in men (P = 0.157). There were no differences between groups in reinfarction rate and target vessel revascularization rate. Multivariate analysis showed age as the only variable independently related to the 6-month mortality, while female gender was not related to the risk of death. The benefit of early PCI is similar in women and men, and any potential referral bias in the use of PCI based on gender differences should be avoided. PMID- 12891600 TI - Coronary stenting in patients with depressed left ventricular function: acute and long-term results in a selected population. AB - Percutaneous coronary angioplasty (PTCA) in patients with depressed left ventricular ejection fraction (LVEF) is associated with increased acute and late mortality; in contrast to plain PTCA, results of stenting in these patients have not been characterized. To assess the current outcome of stenting in patients with LV dysfunction, results from 80 patients procedures were analyzed. Intervention for acute myocardial infarction (MI) was excluded; 21% of patients had unstable angina and 30% had a recent MI. Mean LVEF was 40 +/- 9% (range, 25 45%). Multivessel revascularization was done in 25 patients (31%), with a total of 114 lesions treated. Prophylactic intra-aortic balloon pump was used in only two patients. Angiographic and clinical success was achieved in 79/80 patients (99%). There were no in-hospital deaths, one patient (1%) had a non-Q-wave MI, and no patients required emergency bypass surgery (CABG). All patients completed at least 6 months follow-up (mean, 30 +/- 14 months): 64 patients (80%) remained asymptomatic, 4 (5%) had acute MI, and 5 (6%) died. In-stent restenosis occurred in five patients (6%); of these, three required repeat PTCA, three patients (4%) underwent subsequent elective CABG. Including patients with repeat intervention, 67 patients (84%) are clinically improved; actuarial event-free survival was 87% at 56-month follow-up. Thus, stenting in patients with impaired LVEF is associated with excellent outcome and lower mortality than previously reported for balloon angioplasty alone. Whether coronary stenting may be a therapeutic strategy equivalent to surgery in selected patients needs to be investigated in prospective randomized trials. PMID- 12891601 TI - Stenting in patients with depressed LVEF: expanding the territorial boundaries of PCI. PMID- 12891603 TI - Radial loop and extreme vessel tortuosity in the transradial approach: advantage of hydrophilic-coated guidewires and catheters. AB - Between October 2000 and October 2001, all transradial cases performed by the author necessitating radial and upper arm angiography because of difficult advancement of standard guidewires or catheters were analyzed retrospectively. Fourteen of 594 (2.4%) transradial cases met the study criteria. Radial loops or stenosis and tortuosity in the subclavian or innominate artery were responsible for the difficult access. Several examples of patients with access problems are presented. A technique using a hydrophilic-coated guidewire and a new hydrophilic coated guiding catheter is described. PMID- 12891602 TI - Clinical outcomes following stent implantation in internal mammary artery grafts. AB - We evaluated our experience with percutaneous coronary intervention (PCI) of internal mammary artery (IMA) grafts. From the institution's database we identified 288 patients with 311 IMA lesions. Of these, 82 (26.4%) had stents placed during PCI. Angiographic success was 92%. Mortality at 1 month was 1.7%, myocardial infarction (MI) 15.7%, and target lesion revascularization (TLR) 0.4%. Cumulative 1-year event rates were mortality 6.4%, MI 20.4%, and TLR 8.0%. TLR rates were significantly higher in the stented lesions than lesions treated with angioplasty alone (19.2% vs. 4.9%; P = 0.004). The higher TLR rate in stented lesions was most apparent at the anastomotic site (25.0% vs. 4.2%; P = 0.006). Percutaneous revascularization of IMA grafts can be performed safely with high procedural success and excellent short- and long-term results. Stenting, particularly at the anastomotic site, was associated with significantly greater rates of TLR than angioplasty alone. PMID- 12891604 TI - Direct stenting with a combined intravascular ultrasound-coronary stent delivery platform: a feasibility trial. AB - IVUS is considered as the most accurate tool for the assessment of optimal stent deployment. Direct stenting has shown to be a safe, efficient, and resource saving procedure in selected patients. In a prospective 1-month feasibility trial, a new combined IVUS-coronary stent delivery platform (Josonics Flex, Jomed, Helsingborn, Sweden) was evaluated during direct stenting in consecutive patients considered eligible for direct stenting. The feasibility endpoint was successful stent deployment without any clinical adverse event, while the efficacy endpoint was strategic adaptation according to standard IVUS criteria for optimal stent deployment at the intermediate phase (after a result considered angiographically optimal) and at the end of the intervention (after optimization according to IVUS standards). A total of 16 patients were successfully treated with this device without any major clinical complication. At the intermediate phase, optimal stent deployment was achieved in four patients only, while at the end only one patient had nonoptimal IVUS stent deployment. In particular, the minimal in-stent cross-section area increased from 6.3 +/- 1.2 to 8.3 +/- 2.5 mm(2). These preliminary data demonstrate the feasibility of direct stenting with a combined IVUS-stent catheter in selected patients and confirm the results from larger randomized trials on the impact of IVUS on strategic adaptations during coronary stent placement. PMID- 12891605 TI - Safety and effectiveness of stent implantation without predilation for small coronary arteries. AB - The purpose of this study was to examine the safety and effectiveness of stenting without predilation (direct stenting) in small (< 2.5 mm) coronary arteries. Clinical and angiographic data from the multicenter randomized PREDICT trial was examined. Direct stenting was performed on 42 small and 158 large vessels. Predilation was performed prior to stenting for 37 small vessels. In the small vessel group, mean reference vessel diameter was 2.24 +/- 0.19 mm for direct stenting vs. 2.24 +/- 0.22 mm for predilation (P = NS). Mean reference vessel diameter was 3.17 +/- 0.48 mm in the large-vessel group. Device success for direct stenting was similar for small vs. large vessels (90.5% vs. 92.4%; P = NS). At 180 days, a trend toward increased MACE between small vs. large vessels in the direct-stenting group was observed (26.8% vs.16.7%; P = NS), and significantly higher target lesion revascularizations (TLRs) in the small-vessel group (19.5% vs.7.7%; P = 0.03) were observed. In small vessels, differences in device success rate (90.5% vs. 100 %; P = NS) and lesion success rate (100% vs. 100%; P = NS) for direct stenting vs. predilation were not significant. At 180 days, no differences between direct stenting and predilation for MACE (26.8% vs. 29.7%; P = NS) or TLR (22.0% vs. 21.6%; P = NS) were seen in small vessels, although binary angiographic restenosis rate tended to be higher in direct stenting group (38.2% vs. 18.5%; P = NS). Direct stenting of small vessels can be performed safely and successfully in selected cases. Restenosis rates may be higher for small vessel treated by direct stenting compared to predilation. PMID- 12891606 TI - Platelet glycoprotein IIb/IIIa inhibition with eptifibatide: prolongation of inhibition of aggregation in acute renal failure and reversal with hemodialysis. AB - We report the time course to the restoration of normal platelet aggregation in three patients who were treated with eptifibatide in the setting of severe renal failure. There was prolonged platelet inhibition in the first patient who developed acute renal failure. The second patient with acute on chronic renal failure developed an intracerebral hemorrhage. Normal platelet aggregation was restored after acute hemodialysis. In a third patient with end-stage renal disease, platelet function returned to the normal range following hemodialysis. There is a prolonged time course to the restoration of normal platelet aggregation in patients with severe renal dysfunction who are treated with eptifibatide. Acute renal hemodialysis may reverse the inhibitory effect of eptifibatide on platelet aggregation in these patients. PMID- 12891607 TI - Disassembly of a rotablator: getting out of a trap. AB - The rotablator burr rarely becomes trapped within calcified lesions. Manual traction can be ineffective and dangerous. We report a case that illustrates a novel technique involving use of a percutaneous snare in conjunction with partial disassembly of the rotablator device to remove a trapped burr without need for open surgical intervention. PMID- 12891608 TI - Kokesi, a Japanese wooden doll. PMID- 12891609 TI - Complex decision-making for percutaneous coronary intervention in a patient with coronary artery bypass grafting: use of FFR in multivessel lesion selection. PMID- 12891610 TI - Percutaneous intervention for unprotected left main disease prior to explantation of a left ventricular assist device. AB - Percutaneous coronary intervention of unprotected left main coronary arterial disease is an alternative to surgical revascularization in selected patients. In this report, a patient with an implanted left ventricular assist device (LVAD) underwent successful coronary intervention prior to its planned removal. The implanted LVAD clearly assisted the technical performance of the intervention. PMID- 12891611 TI - The present is favorable but the future remains grim: hopefully not for so long! PMID- 12891612 TI - Rotaglide-facilitated stent delivery: mission accomplished. AB - Percutaneous coronary interventions are performed with a high success rate, though failures still occur. Inability to deliver stents to the target lesion is the most common cause for failures. We present two cases using Rotaglide applied on the stents to enable delivery, showing that Rotaglide is effective in improving stent delivery. PMID- 12891613 TI - New technique for superior guiding catheter support during advancement of a balloon in coronary angioplasty: the anchor technique. AB - To get superior guiding catheter support, we tried a new method called the anchor technique. By inflating a balloon in a nontarget vessel and holding its shaft with backward force while advancing another balloon, the anchor effect for the guiding catheter could be obtained and it appeared to be helpful for a balloon or a stent to cross the target lesion. PMID- 12891614 TI - Conflicting functional assessment of stenoses in patients with previous myocardial infarction. AB - The utility of fractional flow reserve, absolute and relative flow reserve, and intravascular ultrasound may have an impact on decision-making for percutaneous coronary intervention in patients with previous myocardial infarction and microvascular dysfunction. The role for fractional flow reserve, absolute and relative flow reserve, and intravascular ultrasound is discussed. PMID- 12891615 TI - Assessment of the trackability, flexibility, and conformability of coronary stents: a comparative analysis. AB - The efficacy and safety of coronary stent implantation depend on the mechanical features of these devices when deployed in atheromatous lesions of various morphologies. We evaluated the trackability, flexibility, and conformability of 17 coronary stents using specific mechanical bench tests. The quantifications used a dynamometer for assessment of trackability (maximal strength) and flexibility (stiffness) and a 3D optical gauging machine for assessment of conformability (distance between stent and arterial wall in a curvature). The maximal strength (measuring the trackability) ranged respectively from 0.24 +/- 0.06 and 0.38 +/- 0.03 N (Seaquest) to 1.31 +/- 0.42 and 1.34 +/- 0.35 N (Carbostent), concerning respectively curvatures of 90 degrees (P < 0.0001) and 135 degrees (P < 0.0001). The stiffness (measuring the flexibility) ranged from 0.53 +/- 0.16 (Seaquest) to 1.28 +/- 0.10 N/mm (NIR Royal; P < 0.0001). The mean distance between stent and external curvature (external conformability) ranged from 0.15 +/- 0.06 mm (S7) to 0.57 +/- 0.4 mm (NIR Royal; P < 0.0001). The mean distance between stent and internal curve (internal conformability) ranged from 0.26 +/- 0.13 (S7) to 0.44 +/- 0.12 mm (S670; P < 0.0001). These results may influence the choice of a particular stent adapted to a specific coronary anatomy. PMID- 12891616 TI - Late stent thrombosis in the nonbrachytherapy population: a real phenomenon? PMID- 12891617 TI - Retrograde left ventricular hemodynamic assessment across bileaflet prosthetic aortic valves: the use of a high-fidelity pressure sensor angioplasty guidewire. PMID- 12891618 TI - The VB-1 catheter: a novel catheter for peripheral arterial revascularization. AB - Despite recent advances in catheter-based technologies for peripheral arterial revascularization, adequate guide catheter positioning and support remains a leading procedural challenge. This report describes the adaptation of the Behar Internal Mammary VB-1 catheter for peripheral revascularization procedures in patients with complex anatomy not amendable to other conventional peripheral guide catheters. PMID- 12891619 TI - Preliminary experience with a new 18 mm Amplatzer PFO occluder for small persistent foramen ovale. AB - We report our initial experience with a new 18 mm Amplatzer PFO device for the closure of small PFO. Patients were selected by a semiquantitative sizing procedure using the circle of a diagnostic pigtail catheter instead of a sizing balloon. In all eight patients, the device was easy to implant without problems. Due to its size, it has the potential to minimize the risk of inadvertent atrial erosion or perforation after deployment and thus will enhance the safety of interventional PFO closure. PMID- 12891620 TI - Transcatheter closure of very large (>or= 25 mm) atrial septal defects using the Amplatzer septal occluder. AB - Between June 1999 and September 2002, 45 patients (age, 34 +/- 13 years; mean shunt ratio, 2.6 +/- 0.6) underwent transcatheter atrial septal defect (ASD) closure at our institution with the Amplatzer septal occluder (mean device size, 31.4 +/- 3 mm). Patients were selected by transesophageal echocardiography. The mean ASD dimension was 25.3 +/- 3.7 mm and 33 (73%) patients had deficient anterior rim. Specific procedural details included the use of 13 or 14 Fr introducer sheaths and the right upper pulmonary vein approach if the conventional approach failed. There were two procedural failures, with device embolization in both (surgical retrieval in one, catheter retrieval in one). During follow-up (3-30 months; median, 16 months), one patient (59 years) with previous atrial flutter had pulmonary embolism and was managed with anticoagulation. Two patients developed symptomatic atrial flutter. Fluoroscopy time was 31.6 +/- 19.5 min for the first 22 cases and 19.6 +/- 11.4 min for the rest (P = 0.04). Transcatheter closure of large ASDs is technically feasible but careful long-term follow-up is needed to document its safety. PMID- 12891621 TI - Aortic dissection after stent dilatation for coarctation of the aorta: a case report and literature review. AB - A case of stenting for native coarctation is described in a 65-year-old female with a fatal dissection after implantation. The histology of the aorta in coarctation and in the elderly is described. The experience of stenting in older patients is reviewed and discussed. PMID- 12891622 TI - Catheter intervention for adult aortic coarctation: be very careful! PMID- 12891623 TI - Clinical course and management strategies for hemolysis after transcatheter closure of patent arterial ducts. AB - Residual flows following transcatheter coil or device closure of the patent ductus arteriosus (PDA) can result in hemolysis. Of 611 patients who underwent transcatheter PDA closure at our institution, 5 patients (age, 6-63 years) developed overt hemolysis (after coil occlusion in 4 and Amplazter device closure in 1). All had ducts > 3 mm and residual flows after the procedure. In one patient, hemolysis occurred 3 months after coil occlusion following a period of uncontrolled hypertension. The occurrence of hemolysis correlated significantly with both age as well as duct size (P < 0.00001). Hemolysis was associated with a fall in hemoglobin of 3-6 g/100 ml (n = 3), jaundice (n = 2), and renal failure (n = 1). Hemolysis subsided spontaneously in one patient and four patients required flow elimination. Deploying additional coils in three patients eliminated residual flows. In one patient (after Amplatzer device closure for 12.5 mm duct with aneurysm), flow persisted after 25 additional coils, transient balloon occlusion, and gel foam instillation. Flow elimination was eventually achieved through thrombin instillation after balloon occlusion of the ampulla. All patients recovered completely and were well on follow-up. Although hemolysis after duct occlusion is rare (0.8% in this series), residual flow at the end of the procedure merits careful monitoring. Aggressive elimination of residual flows is often necessary to control hemolysis. PMID- 12891624 TI - Percutaneous stenting of anastomotic stenoses in tubular interposition grafts used to repair aortic coarctation in adults. AB - Stenting is the usual treatment for recoarctation following resection with direct end-to-end anastomosis. We describe for the first time stenting to relieve anastomotic stenoses in tubular interposition grafts, which are used to repair approximately 10% of coarctations. Success in these two adults expands further the spectrum of large conduits that may be relieved of stenosis by stenting. PMID- 12891625 TI - The stent. PMID- 12891626 TI - Effect of a single dose of polychlorinated biphenyls on hepatic cell proliferation and the DNA binding activity of NF-kappaB and AP-1 in rats. AB - Polychlorinated biphenyls (PCBs) are environmental pollutants that, because of their persistence and biomagnification, raise concerns about the health consequences of long-term exposure. PCB mixtures induce hepatocellular carcinomas in rodents, but the mechanism of their promoting activity is not clear. Previous studies have shown that oxidative stress occurs after PCB administration, with the induction of lipid peroxidation and oxidative DNA damage, which may contribute to their promoting activity. In this study, we examined whether the oxidative stress-sensitive transcription factors NF-kappaB or AP-1 were activated by PCBs in the liver. Male Sprague-Dawley rats were injected i.p. with corn oil, 2,2',4,4',5,5'-hexachlorobiphenyl (PCB-153, 30, 150, or 300 micromol/kg), 3,3',4,4'-tetrachlorobiphenyl (PCB-77, 30, 150, or 300 micromol/kg), or both PCBs (each 30 or 150 micromol/kg). Rats were euthanized 2, 6, or 24 h, or 2, 6, and 10 d after the PCB injection. Electrophoretic mobility shift assays (EMSAs) were performed to determine NF-kappaB and AP-1 DNA binding activities. The highest NF kappaB DNA binding activity was observed in rats receiving higher doses of PCB 153 (150 and 300 micromol/kg), with peak activation occurring 2 d after injection. AP-1 activation was not detected at any timepoint. Hepatocyte proliferation, as measured by the labeling index, was increased only in groups receiving the highest dose of PCB-153 or the combination of two PCBs (150 micromol/kg each) at day 2, and not by any other PCB treatment at any timepoint. These results show that PCB-153, but not PCB-77, can induce hepatocyte proliferation and hepatic NF-kappaB activation after a single dose. PMID- 12891627 TI - Gene expression changes following androgen receptor elimination in LNCaP prostate cancer cells. AB - We have shown recently that inhibition of androgen receptor (AR) expression with an antisense AR oligonucleotide (ODN) inhibits LNCaP prostate tumor cells in vitro as well as in vivo. In this study, we investigated gene expression changes that occur after AR signaling blockade, either through AR elimination by antisense treatment or through complete androgen receptor inhibition by androgen deprivation combined with the antiandrogen bicalutamide, in order to search for genes that are directly or indirectly regulated through the AR. Gene expression changes were investigated with cDNA NIH 10K gene microarrays in response to treatment over 48 h. Expression of selected genes was further analyzed by real time reverse transcriptase (RT)-polymerase chain reaction (PCR), Western blotting, and radioimmunoassay. A comparison of antisense-treated and androgen deprived cells revealed several concordances such as significant downregulation of prostate-specific genes, cell-cycle regulatory genes, genes of the cholesterol biosynthesis pathway, and several cytoskeletal genes. However, there were also several genes that were differentially regulated. Among the genes that were exclusively changed by treatment with the antisense AR ODN were the insulin-like growth factor binding protein 2 (IGFBP2) and the phosphatidylinositol-4-phosphate 5-kinase type I alpha (PIP5KIA). On the other hand, complete androgen receptor blockade induced changes in the expression of the prostate overexpressed gene 1 and the S100 calcium binding protein P. In summary, we identified a cohort of interesting genes whose expression was highly affected by elimination of the AR in LNCaP prostate cancer cells. Further investigations are warranted to clarify their role in the AR signaling pathway and their susceptibility as a target for the treatment of prostate cancer. PMID- 12891628 TI - Reduced gap junctional intercellular communication and altered biological effects in mouse osteoblast and rat liver oval cell lines transfected with dominant negative connexin 43. AB - Gap junctional intercellular communication (GJIC) maintains normal growth and differentiation of cells in a tissue. The intercellular molecules traversing gap junctions are largely unknown, but the molecular weight (MW) cutoff is normally 1200 Da. No differences in dye transfer were observed in normal or vector controls of WB-F344 rat liver epithelial or mouse osteoblastic MC3T3-E1 cells with either Lucifer Yellow (LY) with a MW of 457 Da (LY-457) or LY with a MW of 649 Da (LY-649). Transfection of a dominant negative-connexin 43 (Cx43) gene decreased GJIC (>50%) when LY-649 was used, however, normal GJIC was observed in both cell lines when LY-457 was used. Therefore, the MW cut off in these clones was considerably less than the wild type. The dominant negative clones of the MC3T3-E1 cells exhibited over 90% less alkaline phosphatase (ALPase) activity and calcium deposition after the induction of differentiation. Similarly, dominant negative Cx43 inhibited gene expression of ALPase and bone sialoprotein but not osteocalcin in MC3T3-E1. WB-F344 cells normally exhibit a biphasic response to 12 O-tetradecanoylphorbol-13-acetate (TPA) where inhibition of GJIC recovers after 2 h, but the dominant negative clones showed no recovery from inhibition of GJIC by TPA. Dominant negative Cx43 also inhibited the formation of network-like structures by WB-F344 cells on Matrigel. These results demonstrate that the dominant negative gene transfected into cell types containing the wild-type connexins result in diminished channel sizes, thus allowing the determination of whether distinct biological endpoints, i.e., differentiation, are dependent upon either small or high MW intercellular signals. PMID- 12891629 TI - Polymorphisms of estrogen receptor alpha in prostate cancer. AB - Estrogen receptor (ER) alpha polymorphisms have been shown to be involved in the oncogenesis of several organs. We hypothesize that polymorphisms of the ERalpha gene are risk factors for prostate cancer. The genotypic distributions of six different loci (codons: 10 T-->C, 87 G-->C, 243 C-->T, 325 C-->G, 594 G-->A, and intron 1 C-->T) of the ERalpha gene were analyzed in prostate cancer tissues. The DNA from 115 cases of prostate cancer (Japanese population) was analyzed by sequence-specific polymerase chain reaction (PCR) and direct sequencing to determine the genotypic and allelic frequencies of the six different polymorphic loci of ERalpha. The relative risk of variant genotype was calculated by comparison with 200 healthy controls. Results of this study showed that the frequency of the variant genotype (C/C) on codon 10 was significantly higher in prostate cancer patients. The odds ratio (OR) was calculated as 3.26 compared to wild-type (T/T) with a 95% confidence interval (CI) of 1.58-6.73. Allele frequency at codon 10 also differed between groups. No association was found between codon 10 polymorphism and the stage of cancer. Polymorphism was not observed in codon 87, and frequencies of genotypes and alleles at other loci (intron 1, codons 243, 325, and 594) were not statistically different between cancer and controls. The present study suggests that polymorphism in codon 10 of ERalpha may be a risk factor for prostate cancer. These results are important in understanding the role of ERalpha polymorphism in the pathogenesis of prostate cancer. PMID- 12891630 TI - Microarray analysis of xenograft-derived cancer cell lines representing multiple experimental models of human prostate cancer. AB - Expression analysis of 7129 transcripts was carried out in five human prostate cancer cell lines derived from orthotopic xenografts after one to five passages in nude mice and primary cultures of human normal prostate epithelial (NPE) cells. These experiments identified a consensus class of 214 genes (43 up- and 171 downregulated transcripts), expression of which was altered at least twofold in the same direction in all the cell lines relative to NPE cells. To validate the relevance of altered expression behavior of these genes for human prostate cancer, their expression pattern was evaluated in multiple additional experimental and clinical settings. Expression of 170 of these 214 genes (79%) was altered in the same direction in vivo in experimental human prostate tumors in mice. Similarly, the expression of 151 of the 214 genes (71%) was altered in the same direction in M12 cells, a variant of an SV40 large T antigen transformed normal human prostate epithelial cell line selected for increased malignancy in vivo. In clinical samples of human prostate tumors, the changes in transcript expression levels of majority of these genes (85% of downregulated and 76% of upregulated transcripts) are consistent with alterations of their expression pattern in xenograft-derived cancer cell lines. These results imply that the expression pattern of a large class of genes is consistently altered in multiple experimental models and clinical samples of human prostate cancer and underscore the potential relevance of the xenograft models and cell lines derived from them for expression analysis studies relevant to human cancer. PMID- 12891631 TI - Introduction: Stigma and discrimination against older people with mental disorders. PMID- 12891632 TI - Reducing stigma and discrimination against older people with mental disorders: a technical consensus statement. AB - This technical consensus statement is jointly produced by the Old Age Psychiatry section of the World Psychiatric Association and the World Health Organization, with the collaboration of several NGOs and the participation of experts from different Regions. It is intended to be a tool for (i) promoting debate at all levels on the stigmatisation of older people with mental disorders; (ii) outlining the nature, causes and consequences of this stigmatisation; and (iii) promoting and suggesting policies, programmes and actions to combat this stigmatisation. PMID- 12891633 TI - Stigma and discrimination against older people with mental disorders in Europe. AB - BACKGROUND AND OBJECTIVES: The European Office of the World Health Organization (WHO/EURO) has established a Task Force On Destigmatization. Upon the release of the Technical Statement on Old Age Psychiatry 'Reducing stigma and discrimination against older people with mental disorders', the Task Force run a small survey in two European subregions to investigate the extent of stigma and discrimination with reference to the elderly person affected from a psychiatric disorder in the countries of those subregions. METHODS: WHO/EURO mental health counterparts completed a short questionnaire inquiring on the inclusion of epidemiological studies on the elderly in their country's research program; the time devoted to the teaching to old age psychiatry during psychiatric residence; the degree of coverage of services for the elderly person and their caregivers; and the extent of stigma present in society with regard to three psychiatric disorders. RESULTS: Stigma and discrimination seem to be present both in the health sector and among the public at large. CONCLUSION: The development of effective health and social facilities to support older persons with mental disorders should be a high priority of any strategy to reduce stigma and discrimination. These facilities should coordinate their efforts with those of other sectors of society to reach the highest possible impact. PMID- 12891634 TI - Training in old age psychiatry. PMID- 12891635 TI - Skill-based objectives for specialist training in old age psychiatry. PMID- 12891636 TI - Teaching and training in old age psychiatry: a general survey of the World Psychiatric Association member societies. AB - OBJECTIVE: One of the main goals of the World Psychiatric Association (WPA) section on old age psychiatry is to promote its discipline and consequently to help member societies to promote and develop locally specific medical education programmes in old age psychiatry. In order to target its support to the needs expressed by the member societies, the section conducted a survey within the WPA during the year 2001. METHOD: A questionnaire was sent to each of the 116 WPA member societies in 93 countries. It assessed the level of development of medical education in old age psychiatry in each country, as well as the perceived needs in educational material. RESULTS: Answers were obtained from 50 WPA member societies (response rate 54%) from 48 countries (response rate 52%). The existence of specific old age psychiatry services was reported in 40 (83%) countries, but the discipline is recognized as a subspecialty in only 13 (27%). Formal teaching in old age psychiatry was reported at the undergraduate level in 44 (92%) countries. Specific learning objectives for postgraduate training were reported in 30 (86%) of the countries where the discipline is not yet recognized as a subspecialty. Specific Continuing Medical Education (CME) programs were however reported in only 50% of the countries. Support for the development of educational material was mostly thought to be useful at the postgraduate level. Depression, dementia, psychopharmacology and psychotherapy were among the most frequently cited areas in which educational material might be usefully provided. CONCLUSION: Support for the development of postgraduate training is seen as the most pressing need by WPA member societies. Such support could help to increase the level of recognition of the discipline locally and hence provide more effective support to older people with mental disorders. PMID- 12891637 TI - Opioid treatment for agitation in patients with advanced dementia. AB - BACKGROUND: Some patients with advanced dementia cannot convey the experience of pain verbally and may react to pain with aggressive and agitated behaviors. We hypothesized that unrecognized pain could contribute to agitation and that low dose opioid therapy might reduce agitation by reducing pain. We therefore attempted to determine the effect of opioids on agitation in demented patients. METHODS: We administered placebo for 4 weeks and a long-acting opioid for another 4 weeks to nursing home patients with advanced dementia and severe agitation despite treatment with psychotropic drugs. Patients and study nurses did not know if the medication administered was placebo or opioid. We measured the Cohen Mansfield Agitation Inventory (CMAI) score at baseline and every two weeks. RESULTS: Among 47 patients who entered the study, 25 completed the two phases. The median age for the 25 patients was 85.5 years. Analyses of the data of these 25 patients and of the patients <85 years-old showed no significant differences in agitation level between the placebo and opioid phases. However, among the 13 patients who completed the study and were > or =85 years old, the agitation level at the end of the opioid phase was significantly lower than at the end of the placebo phase (mean change in CMAI score: -6.4; 95% confidence interval (CI): 10.96, -1.8). The decrease in agitation in the patients > or =85 years old persisted after adjusting for sedation. The results remained unchanged when we expanded the analyses to include four > or =85 patients who dropped out of the study after the second week of the opioid phase. CONCLUSION: Low dose, long acting opioids can lessen agitation that is difficult to control in very old (> or =85) patients with advanced dementia. PMID- 12891638 TI - Can IQCODE detect poststroke dementia? AB - BACKGROUND: Little is known about the performance of the Informant Questionnaire on Cognitive Decline in the Elderly (IQCODE) in the screening of post-stroke dementia (PSDE). METHODS: At 3 months after the index stroke, a research assistant administered the IQCODE to relatives of 189 Chinese patients with acute stroke who were consecutively admitted to a general hospital. A psychiatrist, who was blind to the IQCODE scores, interviewed all 189 patients and made DSM-IV diagnosis of dementia, which served as the benchmark for judging the performance of IQCODE in screening PSDE. RESULTS: The optimal cut-off point of IQCODE was 3.40. The sensitivity, specificity, and positive and negative predictive values of IQCODE, and the area under the receiver operating characteristic curve, were 88%, 75%, 33%, 98%, and 0.88, respectively. CONCLUSIONS: When used as a sole instrument, IQCODE does not appear to be useful in screening PSDE in Chinese elderly. PMID- 12891639 TI - Prevalence of dementia in a semi-urban population in Sri Lanka: report from a regional survey. AB - BACKGROUND: The prevalence of dementia in Sri Lanka, which has a rapidly ageing population, is unknown. OBJECTIVE: This study aimed to determine the prevalence of Alzheimer's disease (AD) and other dementias in a semi-urban elderly Sinhala speaking population in Ragama, Sri Lanka. METHODS: The study was conducted in two phases. Phase I: After informed consent 703 subjects aged > or =65 years from the study area (population 15 828) were screened for cognitive impairment using the Sinhalese Mini Mental State Examination. Subjects scoring < or =17 were regarded as suspected dementia cases. Phase II: All subjects who screened positive in phase I were included in phase II for detailed evaluation for dementia according DSM IV and NINCDS-ADRDA criteria which included structured neuropsychiatric assessment, laboratory investigations, an axial CT scan of the brain and an informant interview. RESULTS: In the study sample, 61% were female and 86% were between 65-75 years. 42 subjects screened positive in phase I. Of these, 28 subjects were diagnosed as having dementia, giving an overall prevalence rate of 3.98% (95% Confidence Intervals (CI) =2.6-5.7%). Of these, 20 (71.4%) had probable AD, four had vascular dementia (14.3%), two had mixed (vascular and AD) dementia (7.1%), one had Lewy body dementia, and one had dementia due to syphilis. Greater age, illiteracy and female gender were associated with higher prevalence of dementia. CONCLUSION: Comparison with other community studies performed in North India suggests that dementia prevalence is higher in Sri Lanka. This may be due to regional differences in disease incidence. PMID- 12891640 TI - A UK survey of psychiatric services for older people in general hospitals. AB - BACKGROUND: Psychiatric illness is common in older people in general hospitals, but little is known of the service models operating in the UK, or of the views of old age psychiatrists regarding service provision in this area. We set out to determine the range of UK old age psychiatry service models for older people in general hospital wards, and the opinions of clinicians on future service priorities and development. METHOD: A postal questionnaire survey of old age psychiatrists providing psychiatric services to older people in general hospital wards. RESULTS: 73% of services were provided through a generic, sector-based, consultation psychiatry model. The remaining 27% employed a range of general hospital-based liaison psychiatry services for older people, involved in proactively seeking referrals and educating general hospital staff. Those providing a generic sector-based model were significantly slower at responding to referrals. 89% of respondents were unhappy with their service to older people in general hospital wards, with only 11% preferring the generic sector-based model. Organisational barriers to change identified included the management of mental health care and physical care by different organisations. Training, both of psychiatric staff in this specialist area, and of general hospital staff in the detection and basic management of common psychiatric conditions in the general hospital setting, was felt to be necessary. CONCLUSIONS: The management of co morbid psychiatric and physical illness in older people is an important issue for health services. Old age psychiatrists are unhappy with the prevalent, reactive, consultation-based model, preferring a range of liaison psychiatry models based in the general hospital. The most important barriers to service development in this area were the separate managerial arrangements for psychiatric and physical care services, and a lack of evidence for effective old age psychiatry services in this setting. PMID- 12891641 TI - Gender, kinship and caregiver burden: the case of community-dwelling memory impaired seniors. AB - BACKGROUND: This study examined whether there were gender and kinship (spouse, child, more distant relative) differences in caregiver burden. It further examined the constellation of gender and kinship by examining whether adult daughter caregivers experienced greater burden compared to wives, husbands, sons, and other more distant relatives. METHODS: The sample consisted of 305 family caregivers of memory-impaired individuals who were age 70 years or older and resided in non-institutional settings in Arkansas. A cross-sectional design was employed using validated measures to assess both the memory-impaired elders' and family caregivers' self-reported physical and memory status. RESULTS: After controlling for the age and health status characteristics of the memory-impaired elder, sociodemographic and health status characteristics of the family caregiver, and the caregiver coping response (measured by the sense of coherence), multiple regression analyses found kinship, but not gender differences in caregiver burden. Adult children experienced more caregiver burden than more distant relatives. There were no significant differences in caregiver burden between adult children and spouses. Adult daughters had greater caregiver burden scores compared to more distant relatives, but had comparable scores to wives, sons, and husbands. Other significant correlates of burden included caregiver personal characteristics (age and ethnicity) and the sense of coherence. CONCLUSIONS: The study discusses the practice implications of adult children and adult daughters' propensity to suffer burden when caring for their memory-impaired parents living in the community. It also discusses the relevance of caregiver personal characteristics and the sense of coherence as correlates of burden. PMID- 12891642 TI - Performance on the CERAD Word List Memory task: a comparison of university-based and community-based groups. AB - BACKGROUND: Evaluation of patients for Alzheimer's disease often compares an individual's performance on cognitive tests to established norms. The purpose of this study was to compare performance on the CERAD Word List Memory tasks in normal controls from an Alzheimer's disease registry and in community volunteers. METHODS: Scores on Word List Memory tasks were evaluated in cognitively intact participants enrolled in a university-based Alzheimer's disease registry (n=103) and in a sample of community volunteers (n=51). Scores for the two samples were also compared with previously published data from registry-based normal controls and from a representative community-based sample. RESULTS: University-based participants outperformed community volunteers, with most marked differences on Delayed Recall and on a Savings score that contrasted immediate to delayed recall. University-based participants performed similarly to previously published scores for normal controls from another university-based Alzheimer's disease registry, while community volunteers were consistent with published scores available from a representative community sample. CONCLUSIONS: Accurate neuropsychological assessment of Alzheimer's disease may require consideration of potentially subtle differences between older adults tested at university centers and those tested in the community. PMID- 12891643 TI - Assessment of health economics in Alzheimer's disease (AHEAD): treatment with galantamine in the UK. AB - OBJECTIVE: To assess the long-term health and economic impact of treating mild to moderate Alzheimer's disease (AD) with galantamine (16 mg or 24 mg per day) compared to no cholinesterase therapy in the UK. METHODS: The long-term costs and outcomes were assessed using a model developed from longitudinal data on a cohort of AD patients. The model predicts the time until patients require full-time care, defined as the consistent requirement for a significant amount of care and supervision each day. Efficacy data were obtained from three clinical trials comparing galantamine with placebo, forecasts were made for ten years. Costs were determined in 2001 British pounds and discounted at 6% per annum, while outcomes such as time to full-time care were discounted at 1.5%. RESULTS: Without pharmacological treatment, patients are expected to incur costs of 28,134 British pounds over ten years, 70% of costs accrue from providing full-time care. Galantamine (16 mg per day) is predicted to reduce the duration of the full-time care state by 12%; approximately five patients need to be treated to avoid one year of full-time care. The ten-year incremental costs per month of full-time care avoided average pound 192 British pounds per patient and 8,693 British pounds per QALY. Savings (1380 British pounds) are predicted for patients who continue treatment beyond six months and whose cognitive function is maintained or improved. Comparable results were estimated for the 24 mg dose. CONCLUSION: In addition to the clinical benefits associated with galantamine treatment, the savings predicted from delaying when full-time care is needed may offset the treatment costs. PMID- 12891644 TI - Negative symptoms in Alzheimer's disease: a confirmatory factor analysis. AB - OBJECTIVE: To examine different conceptual models of negative symptoms in Alzheimer's disease. DESIGN: Confirmatory factor analysis of cross-sectional data. SUBJECTS: Alzheimer patients (n=281) admitted to a psychogeriatric observation ward. MATERIALS: Nurses' Behavioural observation scale for psychogeriatric inpatients (GIP). Global clinical ratings of severity of dementia and depression based on the Cambridge Examination for Mental Disorders of the Elderly-Dutch version (CAMDEX-N). RESULTS: A unidimensional model of dementia fitted the data poorly. Multidimensional models produced better results. In two- and three-factor models negative symptoms were separated from cognitive impairment and mood disturbances. The more severe the memory impairment, the more socially withdrawn patients were. In this sense negative symptoms may have been secondary to cognitive decline. However, no association was found between negative symptoms and mood disturbances. CONCLUSIONS: Negative symptoms are a prominent clinical feature of Alzheimer's disease and they are related to memory impairment but not to mood disturbances. PMID- 12891645 TI - The psychological health and quality of life of older lesbians and gay men: a snowball sampling pilot survey. PMID- 12891646 TI - Treating the dementia of Iris with a cholinesterase inhibitor. PMID- 12891647 TI - Foreword. PMID- 12891648 TI - GABA, gamma-hydroxybutyric acid, and neurological disease. AB - gamma-Aminobutyric acid (GABA) is the primary inhibitory neurotransmitter in the central nervous system. GABA is converted from glutamic acid by the action of glutamic acid decarboxylase (GAD) of which two isoforms exist GAD65 and GAD67. GABA then is broken down, both within the cell and in the synaptic cleft by GABA transaminase to form succinic semialdehyde. In turn, succinic semialdehyde is converted either to succinic acid by succinic semialdehyde dehydrogenase or into gamma-hydroxybutyric acid (GHB) by succinic semialdehyde reductase. Because GABA modulates the majority of inhibition that is ongoing in the brain, perturbations in GABAergic inhibition have the potential to result in seizures. Therefore, the most common disorder in which GABA is targeted as a treatment is epilepsy. However, other disorders such as psychiatric disease, spasticity, and stiff person syndrome all have been related to disorders of GABAergic function in the brain. This review covers the roles of GABAergic neurotransmission in epilepsy, anxiety disorders, schizophrenia, stiff-person syndrome, and premenstrual dysphoric disorder. In the final section of this review, the GABA metabolite GHB is discussed in terms of its physiological significance and its role in epilepsy, sleep disorders, drug and alcohol addiction, and an inborn error of GABA metabolism, succinic semialdehyde dehydrogenase deficiency. PMID- 12891649 TI - The lumbar puncture for diagnosis of pediatric neurotransmitter diseases. AB - The investigation of infants and children with suspected pediatric neurotransmitter diseases affecting serotonin and catecholamine metabolism is complicated because the measurement of metabolites in peripheral fluids is generally uninformative. Disorders that affect catecholamine (dopamine and norepinephrine) and serotonin neurotransmission, and that do not present with hyperphenylalaninemia, require that a lumbar puncture be performed and that specific metabolites be assessed in the collected cerebrospinal fluid. This review will discuss the disorders affecting catecholamine and serotonin biosynthesis, sample collection and handling, diagnostic methods and expected profiles, problems with diagnosis, and as yet to be described conditions that might be detected using current diagnostic methodologies. PMID- 12891650 TI - Approach to the diagnosis of neurotransmitter diseases exemplified by the differential diagnosis of childhood-onset dystonia. AB - We present our approach to the diagnosis of pediatric neurotransmitter diseases exemplified by the differential diagnosis of children presenting with dystonia. This approach is based upon the primary aim of early diagnosis of treatable conditions and the need for a logical series of investigations. We have tried to be comprehensive with our coverage but are aware that "new" pediatric neurotransmitter diseases continue to be delineated and that, similarly, a proportion of children presenting with dystonia remain undiagnosed. If this is the case, all of the investigations suggested here may need to be performed regardless of age and presentation. However, of more value is a careful clinical reevaluation. PMID- 12891651 TI - Magnetic resonance spectroscopy of neurotransmitters in human brain. AB - Magnetic resonance spectroscopy (MRS) is a noninvasive method that permits measurement of the concentration of specific biochemical compounds in the brain and other organ systems in precisely defined regions guided by MR imaging (MRI). Recently, MRS methods have been developed to measure specific neurotransmitters in the brain. More advanced MRS methods have been developed to measure the synthesis rates and turnover of specific neurotransmitters. These turnover rates can provide measures of brain metabolism similar to radioisotope techniques. Also, investigations of the relationship of brain metabolism and specific neurotransmitter systems are now possible using MRS. Here, we review the MRS techniques and studies of neurotransmitters in the human brain. A discussion of the potential use of these techniques in the context of certain pediatric neurotransmitter disorders will be presented. PMID- 12891652 TI - Autosomal dominant guanosine triphosphate cyclohydrolase I deficiency (Segawa disease). AB - Autosomal dominant guanosine triphosphate cyclohydrolase I (GCH-I) deficiency (Segawa disease) is a dopa-responsive dystonia caused by mutation of the GCH-I gene located on 14q22.1-q22.2. Neurohistochemical examination revealed a decrease of the tyrosine hydroxylase protein as well as its activity in the striatum and decrease of dopamine content, particularly in its ventral portion rich in D1 receptors (striatal direct pathways). Neuroimaging, clinical neurophysiological, and biochemical studies showed preservation of the structure and function of the terminal of the nigrostriatal DA neuron. Clinical neurophysiological studies showed no progressive decrement of DA activities. As the enzymatic activity of pteridine metabolism is highest in the early developmental course, it may modulate dopamine receptors maturing early in the developmental course. Its product, tetrahydrobiopterin, has higher affinity to tyrosine hydroxylase among hydroxylases. Thus, partial deficiency of tetrahydrobiopterin caused by heterozygous mutation of the GCH-I gene decreases dopamine activity rather selectively. This affects the DA receptors that mature early and demonstrates characteristic symptoms age-dependently along with the developmental decrement of the tyrosine hydroxylase activities at the terminals and the maturational processes of the projecting neurons of the basal ganglia. A difference in the ratio of mutant/wild-type GCH-I mRNA that depends on the locus of mutation may explain intrafamilial and interfamilial variation of phenotype. PMID- 12891653 TI - The hph-1 mouse: a model for dominantly inherited GTP-cyclohydrolase deficiency. AB - Dominantly inherited guanosine triphosphate (GTP)-cyclohydrolase deficiency, otherwise known as Segawa's disease or dopa-responsive dystonia, has a wide spectrum of phenotypic expression ranging from asymptomatic to very severe. Penetrance is more frequent in women as compared with men, and there is a variable occurrence of diurnal variation in symptom intensity. Biochemical characterization of the disease has demonstrated lower cerebrospinal fluid levels of tetrahydrobiopterin (BH4), neopterin, and homovanillic acid and low levels of tyrosine hydroxylase protein in the striatum. To investigate the pathophysiology, we have begun to characterize biogenic amine and BH4 metabolism in the GTP cyclohydrolase deficient hph-1 mouse. The data show low brain levels of BH4, catecholamines, serotonin, and their metabolites together with low levels of tyrosine hydroxylase protein within the striatum. The hph-1 mouse therefore provides a good model system in which to study the human disease. PMID- 12891654 TI - Aromatic L-amino acid decarboxylase deficiency: overview of clinical features and outcomes. AB - In this paper, we provide a brief update of diagnostic considerations and biochemical phenotype in L-amino acid decarboxylase deficiency. We review clinical features and outcome data in 11 affected patients, including 7 previously unreported cases. All had onset of the characteristic movement disorder by 6 months of age. The phenomenology of the movement disorder is identical to that previously reported, and includes intermittent oculogyric crises and limb dystonia, generalized athetosis, and impaired voluntary movement in all patients. Autonomic dysfunction is characterized by a significant impairment of sympathetic regulation of heart rate and blood pressure, as documented via detailed studies with spectral analysis techniques in two patients. Functional clinical outcomes as a group remain poor, in spite of a variety of attempted treatment interventions, with marked impairment in motor abilities as well as in speech and communication; however, outcome was quite variable from patient to patient and covered a broad spectrum of neurological disability. Much further work remains to identify and refine the best treatment options for patients with L-amino acid decarboxylase deficiency. PMID- 12891655 TI - Tyrosine hydroxylase deficiency causes progressive encephalopathy and dopa nonresponsive dystonia. AB - Tyrosine hydroxylase (TH) is the key enzyme in the biosynthesis of the catecholamines dopamine, epinephrine, and norepinephrine. Recessively inherited deficiency of TH was recently identified and incorporated into recent concepts of genetic dystonias as the cause of recessive Dopa-responsive dystonia or Segawa's syndrome in analogy to dominantly inherited GTP cyclohydrolase I deficiency. We report four patients with TH deficiency and two with GTP cyclohydrolase I deficiency. Patients with TH deficiency suffer from progressive infantile encephalopathy dominated by motor retardation similar to a primary neuromuscular disorder, fluctuating extrapyramidal, and ocular and vegetative symptoms. Intellectual functions are mostly compromised. Prenatally disturbed brain development and postnatal growth failure were observed. Treatment with levodopa ameliorates but usually does not normalize symptoms. Compared with patients with dominantly inherited GTP cyclohydrolase I deficiency, catecholaminergic neurotransmission is severely and constantly impaired in TH deficiency. In most patients, this results not in predominating dystonia, a largely nondegenerative condition, but in a progressive often lethal neurometabolic disorder, which can be improved but not cured by L-dopa. Investigations of neurotransmitter defects by specific cerebrospinal fluid determinations should be included in the diagnostic evaluation of children with progressive infantile encephalopathy. PMID- 12891656 TI - Vigabatrin and newer interventions in succinic semialdehyde dehydrogenase deficiency. AB - Succinic semialdehyde dehydrogenase (SSADH) deficiency is a rare disorder characterized by an inborn error of the catabolism of the inhibitory neurotransmitter GABA. Because of the deficiency of SSADH, the final enzyme of the GABA degradation pathway, the substrate, succinic semialdehyde, is shunted towards production of 4-hydroxybutyric acid (gamma-hydroxybutyric acid). Elevations of gamma-hydroxybutyric acid can be detected in the physiologic fluids of patients with SSADH deficiency, and forms the mainstay of diagnosis. The clinical features of SSADH deficiency include nonspecific neurologic manifestations such as mental retardation/developmental delay, absent speech, hypotonia, nonprogressive ataxia, features of autism or pervasive developmental delay, developmental language delay (dyspraxia, receptive, and expressive delays), and occasionally, seizures. Although the metabolic pathway has been established, it is not known whether insufficient GABA and/or excess gamma hydroxybutyric acid contribute to the disease phenotype. Pharmacological therapy in patients with this disorder has been limited to vigabatrin, an anticonvulsant that blocks GABA transaminase. This review will discuss therapeutic options in SSADH deficiency, on the basis of patient experience, and preliminary work using a murine model. Finally, a discussion of adjunctive therapies will be included. PMID- 12891657 TI - Succinic semialdehyde dehydrogenase deficiency in children and adults. AB - Succinic semialdehyde dehydrogenase deficiency is a rare disorder of the degradation pathway of gamma-aminobutyric acid. The disorder is detected when 4 hydroxybutyric aciduria is present on urine organic acid analysis, and is subsequently confirmed by enzyme measurement on leucocytes. The disorder has been identified in approximately 350 individuals worldwide. We review the clinical features in 60 patients. The most common characteristics are developmental delay maximally involving expressive language, hypotonia, mental retardation, ataxia, and behavioral problems. Seizures occur in approximately half of patients, and include tonic-clonic, absence, and myoclonic seizures, including status epilepticus. Electroencephalographic findings are background slowing and generalized and focal epileptiform discharges. Magnetic resonance imaging typically reveals increased T2-weighted signal of the globus pallidus bilaterally, with variable involvement of white matter and the cerebellar dentate nucleus. Preliminary human cerebrospinal fluid measurements are consistent with neurometabolic aberrations documented in the murine animal model, with elevations in gamma-aminobutyric acid, gamma-hydroxybutyrate, and homocarnosine, and low glutamine. Succinic semialdehyde dehydrogenase deficiency may be an underrecognized neurometabolic disorder with a nonspecific and wide phenotypic spectrum, and carries implications for a comprehensive fundamental understanding of interrelations between multiple neurotransmitter systems. PMID- 12891658 TI - Murine succinate semialdehyde dehydrogenase deficiency. AB - Inherited succinic semialdehyde dehydrogenase (SSADH) deficiency (gamma hydroxybutyric aciduria) is one of the few neurogenetic disorders of GABA metabolism, and one in which tonic-clonic seizures associate with increased central nervous system GABA and gamma-hydroxybutyrate (GHB). To explore pathomechanisms and develop new preclinical treatment approaches, we developed a murine knockout model of SSADH deficiency. In the absence of intervention, SSADH( /-) mice suffer 100% mortality at week 3 to 4 of life from generalized tonic clonic seizures. In this report, we summarize earlier studies indicating disruption of the GABA/glutamine axis in SSADH(-/-) mouse brain, effective pharmacotherapeutic approaches, preliminary gene-therapy results, and electrophysiological analyses of mutant mice. We also present new evidence for oxidative stress in SSADH(-/-) mice, significant alterations of dopamine metabolism, and abnormal neurosteroid levels in brain, potentially implicating the GABA(A) receptor in pathogenesis. In SSADH deficiency, the accumulation of two neuroactive species, GABA and GHB, is significant because GABA is one of the earliest transmitters expressed in mammals, with key roles in synaptogenesis and myelination, whereas GHB displays a vast array of pharmacological actions. The SSADH(-/-) mouse may represent a useful model in which to explore the effect of GABA and GHB accumulation on central nervous system development and function. PMID- 12891659 TI - Transgenic mouse models of dopamine deficiency. AB - The dopamine system is implicated in several neurological and psychiatric disorders. Genetic mutations or variations that affect dopamine system functions either directly cause or contribute to these disorders, even though other genetic and environmental factors may contribute significantly to some of these disorders as well. Transgenic mice increasingly become important tools in revealing functions of genes that are essential components of the dopamine system as well as in modeling human genetic disorders. We have reviewed a comprehensive list of those genes and compared genetic mutations/variations in humans and transgenic mouse models. The significance and limitations of these animal models as well as future directions are discussed. PMID- 12891660 TI - Potential of gene therapy for pediatric neurotransmitter diseases: lessons from Parkinson's disease. AB - Gene therapy methods have continued to develop rapidly, and many initial limitations that hampered clinical application have been overcome. Thus serious consideration of clinical application of gene therapy is warranted for selected disorders in which the pathogenesis is well defined. Parkinson's disease has been the most extensively studied target of gene therapy for central nervous system disorders and shares many features with pediatric neurotransmitter diseases. Neurotransmitter replacement therapy using catecholamine-synthesizing genes and delivery of neurotrophic factors such as glial cell line-derived neurotrophic factors has been successful in animal models of Parkinson's disease. Application of gene therapy for pediatric neurotransmitter diseases will require delineating the optimal set of genes to correct the consequences of the deficiencies. The optimal anatomical targets and proper timing of the gene replacement must be understood. Safety of gene therapy vehicles and the ability to regulate gene expression will be essential for eventual clinical application. PMID- 12891663 TI - Cyclooxygenase-2 and stroke: the long and short of it. PMID- 12891664 TI - Alzheimer's disease: genetic evidence points to a single pathogenesis. PMID- 12891665 TI - Chronically mad as a hatter: anticholinergics and Alzheimer's disease pathology. PMID- 12891666 TI - Toward a molecular neuropsychiatry of neurodegenerative diseases. AB - Quantitative neuropsychiatry has provided increasingly precise descriptions of behavioral phenotypes associated with neurodegenerative disorders. Degenerative diseases of the brain are disturbances of protein metabolism, with failure of protein degredation by the ubiquitin-proteosome system, production of neurotoxic peptide oligomers, and accumulation of intracellular protein deposits. Abnormalities of amyloid beta peptide, alpha-synuclein protein, and hyperphosphorylated tau protein account for more than 90% of degenerative dementias. Functionally related neuroanatomical systems have shared metabolic characteristics and common vulnerabilities to protein dysmetabolism, providing the basis for phenotypes that reflect the underlying proteotype. Patients with alpha-synuclein disorders are particularly prone to hallucinations, delusions, and rapid eye movement sleep behavior disorder. Patients with tauopathies manifest disproportionate disinhibition and apathy, and may exhibit compulsions. Alzheimer's disease is a triple proteinopathy with abnormalities of A-beta, tau, and alpha-synculein leading to a complex behavioral phenotype. This molecular approach to neuropsychiatry may assist in understanding the mechanisms of degenerative diseases, provide insight into the pathophysiology of neuropsychiatric symptoms, and contribute to monitoring disease-modifying therapies. PMID- 12891667 TI - Neuronal overexpression of cyclooxygenase-2 increases cerebral infarction. AB - Increases in COX-2 enzymatic activity and prostaglandin production have been associated with neuronal injury in both acute and age-related degenerative neurological diseases. In this study, we tested the effects of increased COX-2 activity in a model of transient focal ischemia using a transgenic mouse model in which human COX-2 is constitutively expressed selectively in neurons of the striatum, cerebral cortex, and hippocampus. These COX-2 transgenic mice harbor elevated levels of PGE(2) that are 10-fold higher than nontransgenic levels. A significant increase in infarct volume was observed after middle cerebral artery occlusion with 4 days of reperfusion in COX-2 transgenic mice as compared with nontransgenic littermates. Pretreatment of nontransgenic mice with the selective COX-2 inhibitor SC58236 resulted in a significant reduction of infarct volume in nontransgenic mice, consistent with previous pharmacological studies. However, transgenic COX-2 mice treated with SC58236 did not show a significant reduction. This suggests that chronic increases in COX-2 expression and enzymatic activity, which can occur in aging and in pathological states characterized by oxidative stress and chronic inflammatory processes, can lead to downstream cellular changes that have a negative impact on neuronal survival in cerebrovascular disease. PMID- 12891668 TI - Apolipoprotein Eepsilon4 modifies Alzheimer's disease onset in an E280A PS1 kindred. AB - We previously have identified a large kindred from Colombia in which Alzheimer's disease (AD) is caused by the E280A presenilin 1 (PS1) mutation. The objective of this study was to examine whether environmental and genetic factors are responsible for variation in the phenotypic expression of the E280A PS1 mutation. We genotyped coding and promoter polymorphisms of the APOE gene in carriers of the E280A PS1 mutation. Kaplan-Meier product-limit and Cox proportional hazard models were used in the statistical analyses. DNA was available from 114 carriers of the E280A PS1 mutation, including 52 subjects with AD. APOE epsilon 4 allele carriers were more likely to develop AD at an earlier age than subjects without the epsilon 4 allele (hazard ratio, 2.07; 95% confidence interval, 1.07-3.99; p = 0.030). Subjects with low education were more likely to develop AD later than those with higher education (hazard ratio, 0.476; 95% confidence interval, 0.26 0.87). Low educational level was associated with rural residence (p < 0.001). Promoter APOE variants did not influence either the onset or the duration of the disease. This study is the first to our knowledge to demonstrate that genetic and environmental factors influence age of onset in a kindred with a familial AD mutation. PMID- 12891669 TI - Alcohol consumption and the incidence of Parkinson's disease. AB - Cigarette smoking and caffeine consumption are associated with a decreased incidence of Parkinson's disease (PD). This inverse association may result from neuroprotective effects of cigarette smoke and caffeine, or from a disinclination of future PD patients to engage in habituating behaviors. We investigated the association between consumption of alcoholic beverages, another potentially habituating behavior, and risk of PD in two large prospective cohorts Nurses' Health Study, and Health Professionals' Follow-up Study. We detected 415 new cases of PD during follow-up. Compared with nondrinkers at baseline, the relative rate (95% confidence interval) was 1.0 (0.8-1.3) for drinkers of less than 5 gm/day, 1.0 (0.8-1.4) for 5 to less than 15 gm/day, 1.1 (0.8-1.6) for 15 to less than 30 gm/day, and 0.7 (0.5-1.2) for 30 gm/day or more (p for trend = 0.45). Consumption of wine or liquor was not associated with the incidence of PD. Compared with those who consumed beer less than once per month, the relative rate (95% confidence interval) was 0.7 (0.5-0.9) for one to three drinks of beer per month, and 0.7 (0.5, 0.9) for one or more drinks of beer per week. The risk of PD was similar in individuals who usually consume moderate amounts of alcohol and in abstainers. PMID- 12891670 TI - How much phenotypic variation can be attributed to parkin genotype? AB - To establish phenotype-genotype correlations in early-onset parkinsonism, we have compared the phenotype of a large series of 146 patients with and 250 patients without parkin mutations. Although no single sign distinguished the groups, patients with mutations had significantly earlier and more symmetrical onset, dystonia more often at onset and hyperreflexia, slower progression of the disease, and a tendency toward a greater response to levodopa despite lower doses. After forward stepwise multiple logistic regression analysis, dystonia at onset and brisk reflexes were not longer significantly different but were correlated with age at onset rather than the presence of the parkin mutation. Age at onset in carriers of parkin mutations varied as did the rate of progression of the disease: the younger the age at onset the slower the evolution. The genotype influenced the phenotype: carriers of at least one missense mutation had a higher United Parkinson's Disease Rating Scale motor score than those carrying two truncating mutations. The localization of the mutations was also important because missense mutations in functional domains of parkin resulted in earlier onset. Patients with a single heterozygous mutation had significantly later and more asymmetrical onset and more frequent levodopa-induced fluctuations and dystonia than patients with two mutations. PMID- 12891672 TI - Quantification of myelinated endings and mechanoreceptors in human digital skin. AB - We used immunohistochemistry and confocal microscopy applied to fingertip punch biopsy to study glabrous skin innervation in 14 healthy subjects. In addition to epidermal nerve fibers, we quantified mechanoreceptors and their myelinated afferents. Using digital images and dedicated software, we calculated caliber, internodal and nodal length, and G-ratio of the last four internodes of the myelinated endings. In our skin samples, we found a mean density of 59.0 +/- 29.3 myelinated endings per square millimeter with a mean diameter of 3.3 +/- 0.5 microm and an internodal length of 79.1 +/- 13.8 microm. These findings indicate that Abeta fibers undergo drastic changes in their course from the nerve trunk to the target organ, with repeated branching and consequent tapering and shortening of internodal length. Our work demonstrates that skin biopsy can give information on the status of large myelinated endings as well as unmyelinated sensory and autonomic nerves. Since distal endings are primarily involved in distal axonopathy, skin biopsy can be more suitable than sural nerve biopsy to detect early abnormalities. In addition to diagnostic applications, this technique allows clarification of the mode of termination of Abeta fibers and their relationship with mechanoreceptors, leading to relevant electrophysiological speculations. PMID- 12891671 TI - Minocycline and doxycycline are not beneficial in a model of Huntington's disease. AB - Huntington's Disease (HD) is an inherited neurological disorder causing movement impairment, personality changes, dementia, and premature death, for which there is currently no effective therapy. The modified tetracycline antibiotic, minocycline, has been reported to ameliorate the disease phenotype in the R6/2 mouse model of HD. Because the tetracyclines have also been reported to inhibit aggregation in other amyloid disorders, we have investigated their ability to inhibit huntingtin aggregation and further explored their efficacy in preclinical mouse trials. We show that tetracyclines are potent inhibitors of huntingtin aggregation in a hippocampal slice culture model of HD at an effective concentration of 30 microM. However, despite achieving tissue levels approaching this concentration by oral treatment of R6/2 mice with minocycline, we observed no clear difference in their behavioral abnormalities, or in aggregate load postmortem. In the light of these new data, we would advise that caution be exercised in proceeding into human clinical trials of minocycline. PMID- 12891673 TI - Olfactory biopsies demonstrate a defect in neuronal development in Rett's syndrome. AB - Rett's Syndrome (RTT) is a neurodevelopmental disorder resulting from mutation in the mecp2 gene that encodes methyl CpG binding protein 2, a transcriptional repressor. Because this disease primarily affects neurons, tissue is not available during active disease. We used the olfactory system as a model to investigate abnormalities in neuronal development in RTT, because olfactory receptor neurons (ORNs) are replaced throughout life by ongoing postnatal neurogenesis. Thus, even in the adult, the olfactory epithelium contains neurons at various developmental stages. We obtained biopsies of nasal epithelium containing ORNs from RTT patients and age-matched controls to study the status of the neuronal population using antibodies to stage-specific developmental markers. There were no postprocedure complications. Compared with age-matched controls, there were far fewer mature ORNs, as defined by olfactory marker protein expression, and significantly greater numbers of immature neuron-specific tubulin positive ORNs present. In RTT biopsies, olfactory marker protein-positive neurons displayed abnormal structure. These results suggest that dysfunction of MeCP2 results in decreased survival of mature ORNs with a compensatory increase in neurogenesis, or a failure of immature neurons to mature. Our study indicates that olfactory biopsies provide a method to study neuronal developmental diseases in adults and children. PMID- 12891674 TI - Anticonvulsant properties of acetone, a brain ketone elevated by the ketogenic diet. AB - The ketogenic diet (KD), a treatment for drug-resistant epilepsy, elevates brain acetone. Acetone has been shown to suppress experimental seizures. Whether elevation of acetone is the basis of the anticonvulsant effects of the KD and whether acetone, like the KD, antagonizes many different types of seizures, however, is unknown. This study investigated the spectrum of the anticonvulsant effects of acetone in animal seizure models. Rats were injected with acetone intraperitoneally. Dose-response effects were measured in four different models: (1) the maximal electroshock test, which models human tonic-clonic seizures; (2) the subcutaneous pentylenetetrazole test, which models human typical absence seizures; (3) the amygdala kindling test, which models human complex partial seizures with secondary generalization; and (4) the AY-9944 test, which models chronic atypical absence seizures, a component of the Lennox-Gastaut syndrome. Acetone suppressed seizures in all of the models, with the following ED(50)'s (expressed in mmol/kg): maximal electroshock, 6.6; pentylenetetrazole, 9.7; generalized kindled seizures, 13.1; focal kindled seizures, 26.5; AY-9944, 4.0. Acetone appears to have a broad spectrum of anticonvulsant effects. These effects parallel the effects of the KD. Elevation of brain acetone therefore may account for the efficacy of the KD in intractable epilepsy. PMID- 12891675 TI - Frequency and mechanisms of stroke recurrence after cryptogenic stroke. AB - The purpose of this study was to better understand the frequency and mechanisms of stroke recurrence after the stroke with no determined cause (NC). We prospectively studied consecutive patients with acute cerebral infarcts. We divided the patients into five groups (large artery disease [LAD], cardioembolism [CE], small artery disease [SAD], two or more causes [TMC], and NC) and registered recurrent strokes and prognosis for 1 year. Those in the NC group were compared with other subtypes. A total of 204 patients were included; 56 LAD, 22 CE, 62 SAD, 27 TMC, and 37 NC. During follow-up, there were 7 deaths and 31 first recurrent strokes. Patients of the NC group showed a significantly higher rate (30%) of recurrent stroke than those of other subtypes (LAD 16%; CE 14%; SAD 2%), and it was associated with the existence of mild stenosis ( 0.05). By the analysis of the results of visual acuity it could be concluded that it was significantly improved 6 months after the surgery on cyclovertical muscles. This change was more significant in the sample group of patients (p < 0.01) than in the children from the control group (p < 0.05), who were much older. CONCLUSION: Objective refraction in patients was in the range of low values of hypermetropia (+2.00 Dsph--t 3.00 Dsph). Significant improvement of visual acuity occurred 6 months after the surgery performed on vertical muscles of the eye. These results were achieved by the repeated measuring of objective refraction and the precise correction of every new accommodative element. PMID- 12891725 TI - [Clinical course and prognosis in hypoxic-ischemic encephalopathy]. AB - BACKGROUND: Establishing the value of neurological examination, and additional diagnostic methods (ultrasonography and magnetic resonance imaging of the brain) in the diagnosis and prognosis of hypoxic-ischemic encephalopathy and its treatment, tracking the clinical course, and making the prognosis of neurological development in newborn infants with hypoxic-ischemic encephalopathy. METHODS: The group of 40 term newborn infants with suspected intrauterine asphyxia was examined. All the infants were prospectively followed until the 3rd year of age at the Clinic for Neurology and Psychiatry for Children and Youth in order to estimate their neurological development and to diagnose the occurrence of persistent neurological disorders. All the infants were analyzed by their gestational age and Apgar score in the 1st and the 5th minute of life. They were all examined neurologically and by ultrasonography in the first week of life and, repeatedly, at the age of 1, 3, 6, 9, 12, 18, as well as in the 24th month of life. They were treated by the standard methods for this disease. Finally, all the infants were examined neurologically and by magnetic resonance imaging of the brain in their 3rd year of age. On the basis of neurological finding infants were divided into 3 groups: infants with normal neurological finding, infants with mild neurological symptomatology, and infants with severe neurological disorders. RESULTS: It was shown that neurological finding, ultrasonography and magnetic resonance imaging of the brain positively correlated with the later neurological development of the infants with hypoxic-ischemic encephalopathy. CONCLUSION: Only the combined use of these techniques had full diagnostic and prognostic significance, emphasizing that the integrative approach was very important in the diagnosis of brain lesions in infants. PMID- 12891726 TI - [Importance of cytologic examination of urine in the diagnosis of renal transplant function disorders]. AB - BACKGROUND: This paper presents our experience with cytologic examination of urine in diagnosing renal allograft dysfunction. METHODS: The study group included 23 patients with renal allograft dysfunction, selected from 56 patients who underwent renal transplantation. Etiologic diagnosis was made according to the clinical picture, histological findings during allograft biopsy, and cytologic examination of urine. Urine sediment was obtained in cytocentrifuge and was air dried and stained with May Grunwald Giemsa. RESULTS: Out of 23 patients with allograft dysfunction in 18 (78.3%) patient it was caused by acute rejection, and in 5 (8.9%) patients by allograft infarction, cyclosporine nephrotoxicity, acute tubular necrosis and chronic nephropathy. In eighteen patients (78.3%) cytologic examination of urine was pathologic, while in 16 (70%) clinical and histology findings coincided with urine cytology findings. Out of 18 patients with acute allograft rejection in 15 patients cytologic examination of urine coincided with acute rejection. Out of 7 patients with expressed cyclosporine nephrotoxicity, in 5 cytologic examination of urine confirmed the cause of allograft dysfunction, as well as in one of 2 patients with acute tubular necrosis. Cytologic examination of urine indicated parenchymal damage in 2 patients with recurrent disease (membranoproliferative and focal sclerosing glomerulonephritis). In 4 of 5 patients suffering from chronic rejection in a year's monitoring period, urine sediment periodically consisted of lymphocytes, neutrophilic leucocytes, monocyte/macrophages, tubular cells and cylindres, without the predominance of any cell type. In 3 patients allograft dysfunction was caused by infective agents (bacteria, fungus, cytomegalovirus). CONCLUSION: Cytologic examination of urine might be an alternative to histological in diagnosing acute allograft rejection and acute tubular necrosis or nephrototoxicity. Also it might indicate parenchymal disease while the importance of urine cytology in chronic allograft nephropathy needs to be investigated further. PMID- 12891727 TI - [The soldier's quality of life during the years of military service]. AB - Quality of life of soldiers is the ability to participate in professional activities during the military service in accordance with intrapsychic state. The aim of this study was to establish the categories of quality of life, which are important for soldiers' adjustment during the military service. The investigation was performed on the subjects of the two squads of soldiers of Yugoslav Armed Forces, divided into three groups according to the period of investigations: after the first month, after the basic training, and at the end of military service. The applied instruments of the investigation were sociodemographic questionnaire and Quality of Life Scale (QLS). Statistical analysis of the obtained data was made by t-test. Investigation results showed that quality of life of soldiers during the military service depended on intrapsychic state, interpersonal relations, and soldiers' adjustment to common objects and activities, and did not depend on their occupational role. PMID- 12891728 TI - [Fibrinogen as a risk factor in ischemic heart disease]. PMID- 12891729 TI - [Current trends in the development of clinical electromyography]. PMID- 12891730 TI - [Marking the route of lymphatic spread of melanoma and sentinel lymph node biopsy]. PMID- 12891731 TI - [GABA-A receptors: molecular substrate for the development of new anxiolytic agents]. PMID- 12891732 TI - [Fulminant hepatitis B]. AB - Fulminant hepatitis, or fulminant hepatic failure, is defined as a clinical syndrome of severe liver function impairment, which causes hepatic coma and the decrease in synthesizing capacity of liver, and develops within eight weeks of the onset of hepatitis. Several independent factors influence the survival of patients: age, the cause of liver disease, the degree and the duration of encephalopathy in relation to the onset of the disease, and the prevention of complications. Over the years many intensive treatments have been practiced. Liver transplantation is expensive, and patients who survive transplantation require life-long immunosuppression, clinical care and complications management. Without transplantation fulminant hepatitis and hepatic failure might be completely recovered spontaneously, and the patient could expect a normal life. Two cases of fulminant B hepatitis with intensive care treatment, and their survival despite unfavorable prognosis are presented in this paper. The management of patients with fulminant hepatitis required intensive monitoring and therapeutic measures, including corticosteroids. The prognosis for survival without transplantation in fulminant hepatitis is limited by the measures of medical treatment and new specific therapeutic modalities which must be developed through basic research. PMID- 12891733 TI - [Bigeminy--the result of interaction between digoxin and St. John's wort]. AB - A case of an by digoxin under unusual circumstances is reported. An 80-year-old man, previously on long-term digoxin treatment, started consuming St John's wort herbal tea (2,000 ml/daily) because of frequent episodes of depression. After the cessation of consuming herbal tea containing Hypericum perforatum, digoxin poisoning developed in our patient. Electrocardiography revealed nodal bradicardia 36/min and bigeminy. Manifested symptoms were the consequence of interaction between digoxin and Hypericum perforatum which were consumed simultaneously, and the cessation of consuming St John's wort herbal tea afterwards. Therapy was the same as in the standard digitalis poisoning. Consumers of St John's wort combined with medical products are advised not to discontinue tea consumption on their own, without consulting their physician. PMID- 12891734 TI - [Surgical and prosthetic treatment of large mandibular cysts]. AB - This paper presents a combined surgical-prosthetic procedure of reconstructing mandibular bone defect in a 53 year old patient, following enucleation of a mandibular cyst (Cystectomy Partsch II). After a thorough diagnostic evaluation, a surgical procedure was planned with the particular attention to the nature of the disease, patient's condition, size and extension of the cyst, tissue loss, and the possibilities of prosthetic management of a mandibular bone defect with partial postresection dental prosthesis. It is of great importance to point to the significance of teamwork of a maxillofacial surgeon and a specialist in prosthodontics. This kind of cooperation provided very effective and less risky soft tissue, as well as bone tissue regeneration (osteogenesis). The patient's recovery was fast, and he could return to his daily activities and work without significant changes regarding quality of life after surgery and prosthetic treatment. PMID- 12891735 TI - [The founding and development of the Neurology Clinic at the Academy of Military Medicine 1923-2003]. PMID- 12891736 TI - [History of the Academy of Military Medicine: VII--Department of Military Hygiene (part II)]. PMID- 12891737 TI - [Wartime military surgery in Serbia during the first and second Serbian uprising]. PMID- 12891738 TI - [Local staging of hypopharyngeal tumors using ultrasonography]. PMID- 12891739 TI - [Significance of dynamic studies in the diagnosis of thoracic foci formation]. PMID- 12891740 TI - [Unclear ascites]. PMID- 12891741 TI - [Malaria: between deception and hope]. PMID- 12891742 TI - [Use of satellites for public health purposes in tropical areas]. AB - The epidemiological hallmark of the new millennium has been the emergence or recrudescence of transmissible diseases with high epidemic potential. Disease tracking is becoming an increasingly global task requiring implementation of more and more sophisticated control strategies and facilities for sustainable development. A promising initiative involves the use of satellite technology to monitor and forecast the spread of disease. The Health Early Warning System (HEWS) was designed based on successful application of satellite data in food programs as well as in other areas (e.g. weather, farming and fishing). The HEWS integrates data from communications, remote-sensing and positioning satellites. The purpose of this review is to present the main studies containing satellite data on public health in tropical areas. Satellite data has allowed development of more reactive epidemiological tracking networks better suited to increasing population mobility, correlation of environmental factors (vegetation index, rainfall and ocean surface color) with human, animal and insect factors in epidemiological studies and assessment of the role of such factors in the development or reappearance of disease. Satellite technology holds great promise for more efficient management of public health problems in tropical areas. PMID- 12891743 TI - [Indigenous medical assistance in Madagascar (1898-1950)]. AB - The foundations of local health services in Madagascar were laid when the island was taken over by the French in 1896. Medical care was a major priority for the first colonial governor named General Gallieni. Local health services expanded greatly from 1896 to 1950 thanks notably to institutions such as the Tananarive Medical School and Pasteur Institute. These services played a crucial role in the fight against smallpox and bubonic plague. However they were also used for political purposes by both colonial and independent governments. In sum the history of local health services in Madagascar can be described as a battle between political power and scientific knowledge. The overall result is positive for some and controversial for others. PMID- 12891744 TI - [Osseous tuberculosis]. PMID- 12891745 TI - [Zygomycoses and blastomycoses]. PMID- 12891746 TI - [Severe acute respiratory syndrome spreads over the planet]. PMID- 12891747 TI - [Jet lag]. PMID- 12891748 TI - [Management of American cutaneous leishmaniasis. Outcome apropos of 326 cases treated with high-dose pentamidine isethionate]. AB - The purpose of this retrospective analysis was to evaluate the outcome of pentamidine isethionate treatment (4 mg/kg of Pentamidine by the intramuscular route on Days 1 and 3) of cutaneous leishmaniasis in 326 cases that occurred during an outbreak among French military personnel in French Guyana from 1998 to 1999. A great difference was found between the 205 patients treated in French Guyana (series G) and 32 patients treated at the Laveran Military Hospital in Marseille, France (series L). Failure rate, i.e. 25% in series L versus 5% in series G, was significantly correlated with the delay to treatment which was much longer in series L. Extensive rhabdomyolysis was observed in all cases tested: this side-effect has not been reported. Based on these findings and a review of the literature on pentamidine isethionate, the authors recommend prompt treatment using lower doses. Other treatment alternatives for American cutaneous leishmaniasis are also presented including two of the latest developments in the field, i.e., oral treatment using miltefosine and topical treatment using agents such as paromomycine. PMID- 12891749 TI - [Effectiveness of syndromic approach for management of urethral discharge in Senegal]. AB - The World Health Organization has proposed the syndromic approach for management of sexually transmissible diseases (STD) in countries where diagnostic laboratory tests are not consistently available. The purpose of this study was to evaluate the effectiveness of this approach for treatment of ureteral discharge in Senegal. Twenty seven men presenting ureteral discharge underwent two-week treatment using a combination of cotrimoxazole plus tetracycline for suspected gonococcal and a chlamydial infections. Ureteral samples were collected before and after treatment to detect Neisseria gonorrhoeae by culture and Chlamydia trachomatis by direct immunofluorescence and ELISA. Results demonstrated successful treatment of all patients presenting gonococcal and chlamydial infections i.e. 84.6% of cases. Neither germ was detected in 15.4% of cases. Before treatment, Neisseria gonorrhoeae, Chlamydia trachomatis or both were found respectively in 53.9%, 5.1% and 25.6% of samples respectively. Based on these findings we conclude that the syndromic approach was effective in 84.6% of cases but treatment was in adequation with STD biologically documented only with 25.6% of cases. PMID- 12891750 TI - [Quadriceps fibrosis following intramuscular injections into the thigh: apropos of 92 cases at the Ignace Deen Central University Hospital in Conakry]. AB - Analysis of 92 cases of quadriceps fibrosis following intramuscular injection into the thigh showed that the most affected age group was 3- to 7-year-old preschool children with a male predominance (68.7%). The most implicated drug (81.7% of cases) was quinine salts after of mean of 2 injections. Stiffness of the knee was the most outstanding clinical sign. In our series, the efficient technique for prompt functional recovery consisted of lengthening the quadriceps tendon by isolated "Z" plasty (56.5% of cases). Administration of drugs in oral, rectal or intravenous forms is the most effective means of prevention. PMID- 12891751 TI - [Trachoma in Senegal: results of a national survey]. AB - A national survey was conducted to determine the prevalence of trachoma in Senegal between February and July 2000. The methodology consisted of a cross sectional survey based on random sampling with two levels of stratification in five areas or area groups. A total of 12,328 children under 10 years of age and 8753 women over the age of 14 years were examined from 150 clusters (30 clusters per stratum). Excluding the Dakar stratum, the estimated prevalence of active trachoma (TF/TI) and severe trachoma (TI) in children under 10 years of age was 10.8% (IC 95%: 9.2-11.4) and 1.2% (IC 95%: 0.5-2.3) respectively. The corresponding prevalence for the areas around Dakar were 3.3% et 0.3%. Prevalence rates were highest between the ages of 2 and 3 years (17.4%). However these rates concealed major differences between strata since prevalence ranged from 3.3% in Dakar to 14.2% in Thies-Diourbel. Excluding Dakar, the estimated prevalence of entropion/trichiasis and corneal opacity among women over the age of 14 years was 2.6% (IC 95%: 1.9-3.4) and 1.4% (IC 95%: 0.9-1.8). The prevalence of trichiasis was over 1% in every area except Tambacouna and reached 4% in Thies-Diourbel. Trichiasis was observed in 17.1% of women over the age of 70. Based on extrapolation of these data to the whole country, it can be deduced that 272,020 children have active trachoma requiring medical treatment and that 115,000 persons over 14 years of age present trichiasis requiring surgical treatment. PMID- 12891752 TI - [Acute encephalitis concurrent with primary infection by Schistosoma mansoni]. AB - A 24-year-old man returning from a trip to Mali was hospitalized for acute encephalitis and fever in association with acute primary infection by Schistosomiasis mansoni. Bilharziasis was suspected from the epidemiological context and presence of eosinophilia. Diagnosis was confirmed by serological testing. Specific treatment using praziquantel and corticotherapy was successful. Central nervous system involvement attributable to embolization of eggs or ectopic migration of adult worms has been reported in association with chronic Schistosomiasis by S. japonicum or S. mansoni. There have been few reports of acute neuroschistosomiais during the acute primary phase of infestation by S. mansoni. Etiology probably involves immunoallergic mechanisms. PMID- 12891753 TI - [Abdominal aortic aneurysm manifested by hematemesis in a 32 year old patient infected by HIV]. AB - Aortic aneurysm is an uncommon in young patients. The purpose of this report is to describe a case of dissecting aortic aneurysm in a 32-year-old man with human immunodeficiency virus (HIV) infection. The presenting symptom was recurrent massive hematemesis. Diagnosis was suspected on the basis of clinical cardiovascular assessment and confirmed by ultrasonography and computed tomography. The clinical course was complicated by superior vena cava syndrome and kidney failure. Outcome in the absence of surgical treatment was fatal. Perusal of the literature indicates a rising incidence of large vessel disease in young patients with HIV infection. The underlying pathophysiological mechanisms is unclear. Assessment for arterial lesions should be performed in all patients with HIV infection since clinical signs may be misleading. Definitive diagnosis can be achieved by ultrasonography, Doppler ultrasound or computed tomography. PMID- 12891755 TI - [Correlation measures in epidemiology]. AB - When a significant correlation has been noted between the occurrence of a disease and exposure to a risk factor, it becomes necessary to measure the strength of the correlation. This can be done using either relative risk or odds ratio techniques depending on the type of study. The purpose of article this article is to present guidelines for calculation and interpretation of these indicators using examples taken from the Medecine Tropicale journal. PMID- 12891754 TI - [Polyradiculoneuritis and Campylobacter jejuni: clinical and physiopathological aspects]. AB - Several explanations have been proposed to explain the relationship between axonal forms of acute auto-immune inflammatory polyradiculoneuritis and Campylobacter jejuni. The major hypothesis involving molecular imitation is based on the existence of common antigenic determinants (epitopes) in the lipopolysaccharides of the infectious agent and gangliosides, i.e. glycosphingolipides on the surface of the nervous system cells, especially peripheral nervous system cells. The purpose of this literature review is to improve understanding of the rather complex physiopathological mechanisms underlying Guillain-Barre syndrome. PMID- 12891756 TI - [Malaria prophylaxis and treatment: problems, recent developments and perspectives]. AB - Rapid development of significant resistance to antimalarials has been a major force driving research to identify and develop new drugs. Recent progress in this field promises to lead to a much greater range of antimalarial agents. The availability of a broader battery of drugs should provide a partial solution to the dilemma faced by malarial control agencies, i.e., distributing antimalarial drugs as widely as possible without enhancing resistance. Avenues of research for development of new antimalarials include lipid metabolism, degradation of hemoglobin and proteins, interaction with molecule transport, iron metabolism, apicoplasty and signal transduction. Throughout the course of evolution, micro organisms have thwarted traps set by the environment including those designed by man. One can but hope that the different approaches now being implemented on a worldwide bases will overcome the defense mechanisms that Plasmodium have deployed in their long co-evolution with humans and anopheles. PMID- 12891758 TI - Focusing on communication. PMID- 12891757 TI - [Behavior of the young woman faced with contraceptive methods in the Lome community ( Togo)]. PMID- 12891759 TI - Communication disorders in children. PMID- 12891760 TI - A 6-week-old infant with irreducible inguinal hernia. PMID- 12891761 TI - What pediatricians should know about normal language development: ensuring cultural differences are not diagnosed as disorders. AB - The roles and responsibilities of speech-language pathologists and pediatricians have become greater with the changing population demographics in the United States. In some states, the majority of the population belongs to a national cultural minority, eg, New Mexico. Even a state such as Iowa, with only a 5% nonmajority population, has a school-aged population that is almost 10% nonmajority. This growth of diversity is likely to continue. Rather than viewing sensitivity to the influence of culture on language learning and other developmental areas as an "add-on" to a practice, it may be wiser to recognize that approaching all clients with as few assumptions about their behaviors as possible will guarantee nonbiased service delivery for all. Without nonbiased service delivery, incorrect diagnoses and provision of inappropriate therapy become more likely. Fortunately, many resources are available to assist pediatricians and speech-language pathologists in learning about various cultures. Institutional review boards have become more vigilant about the inclusion of a cross-section of subject populations as participants in research studies in addition to protecting the rights of all participants. Funding agencies also have expressed as a priority the inclusion of research subjects from minority populations to add to the information available about the incidence and prevalence of disorders across the range of our potential patients. In a society in which cultural differences are not just defined by race or ethnicity, but by gender, sexual orientation, age, geographic region, and religion, belief systems about disease, disability, and treatment are dynamic entities for health professionals to take into consideration. It is a challenge that speech-language pathologists and pediatricians must meet if they are to provide the best and most appropriate services for their patients. PMID- 12891762 TI - Developmental stuttering. AB - Pediatricians often are the first health care professional parents turn to when they are concerned their child may be stuttering. At this point, the pediatrician can direct parents to a number of available resources for more information and also can provide referral to a speech-language pathologist who has undergone graduate training and received certification in speech-language pathology. PMID- 12891763 TI - Early childhood hearing loss: a frequently overlooked cause of speech and language delay. AB - The explosion of information regarding the genetics of hearing loss, the extraordinary effectiveness of early intervention, and the widespread practice of universal newborn hearing screening make for exciting times for those who serve young children who are deaf or hard-of-hearing and their families. These rapid changes in the knowledge base and practice standards also require the cooperation and help of pediatricians to enable children to take full advantage of available opportunities to optimize development of early communication. PMID- 12891764 TI - Augmentative and alternative communication. AB - With the help of a knowledgeable interdisciplinary team, individuals can find the best AAC system to meet their needs and enrich their quality of life. A new-found ability to communicate often significantly improves a person's overall prognosis and may prevent serious problems in the areas of behavior, learning, reading, and social development. However, because an AAC system is just one component of multimodal communication, it should not inhibit the use of speech by individuals who have some usable or functional speech. It is important to understand that AAC is a process and that an AAC system will evolve over time to meet the changing preferences, dislikes, and abilities of the user as that individual strives to accomplish communication goals. PMID- 12891765 TI - Speech and language development disorders in infants and children with cleft lip and palate. PMID- 12891766 TI - Nurturing a precious commodity. Trustee recruits. AB - Today's trustees have to be sharper and work harder than ever before. Where will the next crop of health care trustees come from? And how can hospital boards find, recruit and grow the increasingly more sophisticated level of talent they need? PMID- 12891767 TI - Trustee workbook 3. Best practices in action: orientation. PMID- 12891768 TI - Avoiding conflict of interest. How boards can withstand close scrutiny. AB - Recent corporate scandals have focused attention on both for-profit and not-for profit boards. To avoid even the appearance of impropriety, boards need to be above reproach--starting with conflict of interest. PMID- 12891769 TI - Community Health 'R' Us. AB - This year's Foster McGaw winner accepts responsibility for its community's health and well-being as a natural part of its mission. And it's making a huge impact. PMID- 12891770 TI - Governance. Conflict of interest: questions and answers for trustees. PMID- 12891771 TI - [Brain central amygdaloid nucleus: cytoarchitechtonics, neuronal organization, connections]. AB - This review describes different aspects of structural and functional organization of amygdaloid central nucleus (CN) in association with the findings indicating its involvement in the development of stress reactions and adaptive behavior in animals. The data on the distribution of neuropeptides, neurotransmitters and neuromodulators in CN are discussed. It is emphasized that CN appears at the earliest stages of amygdaloid formation, undergoes remodeling together with it and reflects the evolution of the whole amygdaloid. The detailed data are presented on the cytoarchitectonic of amygdaloid CN, its heteromorphism and subdivision into zones (subnuclei), based on the different methods of investigation and evaluation criteria. The neuronal organization of CN and its subnuclei is discussed, and the detailed description of neurons of different types is given according to their topography, cell body dimensions and shape, dendrite orientation and branching pattern, organization of spine apparatus and axon structure. The developmental features of amygdaloid CN in mammalian animal and human ontogenesis are considered. The analysis covering the literature data and the results of authors' own investigations indicates that CN functions not only as intraamigdaloid integrative center, but also as one of the major channels providing both afferent and efferent connections of amygdaloid with other brain structures. PMID- 12891772 TI - [Structural and functional organization of the cerebral cortex neurons in rats with various resistance to emotional stress following administration of delta sleep-inducing peptide]. AB - In Wistar rats with different resistance to emotional stress (ES), subjected to stress exposure (SE), the structural organization of neurons in layer V of sensomotor cortex was studied quantitatively in brain sections stained using Nissl's cresyl violet method. One group of animals was injected with delta sleep inducing peptide (DSIP) 1 hr before SE. In rats of control group predisposed to ES, the amount of normochromatic and moderately hypochromatic neurons was decreased as compared to the rats resistant to ES. After SE, normochromatic neurons were not demonstrated. In rats predisposed to ES, the contents of moderately hypo- and hyperchromatic neurons was found to fall dramatically with a simultaneous increase in the number of extremely hypo- and hyperchromatic neurons, ghost cells and ischemically changed cells. After DSIP infusion before SE, ischemically damaged cells were not found in any group, while the degree of shrinkage of extremely hyperchromatic neurons was lower in rats predisposed to ES as compared to rats resistant to ES. It is suggested that brain hypoxia plays an important role in cortex disorganization during ES, whereas DSIP, possesses both antistress and antihypoxic effects. PMID- 12891773 TI - [The structure of internal intraneuronal connections in the area 17 of the cat cerebral cortex]. AB - Spatial distribution of horizontal intrinsic neuronal connections in area 17 of cat cerebral cortex was studied following HRP microion tophoretic injections into the single cortical columns. Cluster analysis of labelled cell distribution in superficial layers was performed in the tangential cortical plane. Area 17 was found to contain 7 +/- 1 clusters each consisting of 1-5 cells. Clusters were arranged in two rows, separated by a distance of 1.2 +/- 0.3 mm. The distance between the centers of the clusters that form the rows was equal to 0.8 +/- 0.3 mm. The spatial characteristics of cell clusters sending axons to cortical column, found in this study, were compared with the known data on the optical imaging of the activity of neurons of orientation and ocular dominance columns of visual cortex. It is suggested that area 17 cortical column receives inputs from cells of 6-8 hypercolumns that have similar ocular dominance and orientation preference. PMID- 12891774 TI - [NO-ergic rat brain commissural neurons in the norm and during opiate administration]. AB - Using histochemical reaction demonstrating NADPH-diaphorase (NADPH-d), the dynamics of NO synthesis was studied in the rat brain raphe nuclei following intravenous injection of morphine hydrochloride. In normal conditions NADPH-d activity was demonstrated in neurons of all raphe nuclei. Acute and chronic administration of morphine in different doses (0.5 mg/kg and 5 mg/kg) was found to inhibit NO-ergic activity of the major part of raphe nuclei neurons. The depression of NADPH-d activity was unequal in different nuclei. The NO-ergic changes are caused by an activation of opiate receptors, as they depend on morphine dose, while the application of opiate antagonist naloxone restores the NO-ergic function of raphe neurons. Formation of tolerance to opiate analgetic effect is accompanied by a significant, though short-lasting increase of NO synthesis activity. It is suggested that the changes in NO-ergic function of raphe neurons may influence brain serotonin balance after opiate administration. PMID- 12891775 TI - [Changes in the ultrastructure of the rat bulbar nuclei caused by kainic acid administration]. AB - The effect of kainic acid (KA) injection into the commissural area of nucleus tractus solitarii (NTS) of rats was studied to evaluate its influence on the ultrastructure of neurons and gliocytes at the site of injection and in the ventrolateral medulla (VM). 30 min after KA injection, in the perikarya of neurons in rostral and caudal NTS and VM the increased number of ribosomes, lysosomes, lipofuscin granules and edema of mitochondria were noted. Giant mitochondria were found in terminals and axons of myelin fibers. 14 days after KA injection, the increased volume of Golgi complex cisterns was found in single neurons in NTS rostral parts, while the neurons of caudal NTS demonstrated pronounced degenerative and destructive changes. During this period, the volume of mitochondria and of vacuoles was increased in dendrites within caudal NTS and VM, where degenerating processes of the nervous cells were found. In gliocytes of caudal NTS, the hypertrophy of cytoplasm was noted with the accumulation of dilated cisterns of endoplasmic reticulum and redistribution of nuclear chromatin. In the same areas of NTS the reaction of endotheliocytes and pericytes, perivascular edema and accumulation of microgliocytes were found. The results obtained indicate the reactive changes in VM in response to intracentral injection of neurotoxin. PMID- 12891776 TI - [The ultrastructure of desmosome-like contacts in mixed synapses of Mauthner cells followed long-term potentiation]. AB - Using electron microscopic morphometry, the effect of long-term potentiation on the structure of transverse fibrillar bridges localized in a cleft of desmosome like contacts in mixed synapses of goldfish Mauthner neurons, was studied. The number of bridges was shown to increase with the augmentation of the potentiation level of electrotonic conductivity. In contrast to control (incubation), the structure of bridges was also changed after potentiation. The paired bridges appeared that could have an arrangement within the cleft different from that one in control. On the basis of the present results in conjunction with earlier findings, the possible actin nature of these bridges was suggested. The bridge seems to represent a channel containing actin, arranged in a pattern similar to that one in nanotubules or plasmodesmas, what can, possibly, explain the stability of bridge structure observed after application of [symbol: see text] D and other extrinsic damaging factors. PMID- 12891777 TI - [Morphological and cytochemical properties of the human glioblastoma T98G cells during the monolayer culture]. AB - The study was carried out with the aim of determination of morphological and cytochemical properties of human glioblastoma T98G cells in monolayer culture. Using light microscopic and immunocytochemical techniques, the glioblastoma cell line studied was shown to have low susceptibility to the effects of environmental factors and changes in culture conditions as well as high proliferative activity. All T98G cells were demonstrated to have positive nuclear reaction for p53 protein. An ability of T98G cells for spontaneous caspase 3-mediated apoptosis was shown using the antibodies against activated caspase 3. The data obtained suggest that glioblastoma T98G cell line could be used as a convenient model for testing the capacity of natural substances and of pharmaceutical agents that are being developed for the selective induction of caspase 3-mediated apoptosis. PMID- 12891778 TI - [Morphological characteristics of Lugaro cells of the cerebellar cortex]. AB - Two types of Lugaro cells--fusiform and triangular--were found at different levels of granular layer in the sections of cat cerebellar cortex, stained with silver nitrate using Golgi-Kopsch method. Their processes are oriented horizontally, vertically or obliquely to the folium axis, while their axons never leave the limits of cerebellar cortex, therefore these cells should be considered as interneurons. The processes of Lugaro cells have very large spatial expansion, due to which these cells form numerous axosomatic and axodendritic connections with all the neurons and fibers of the cerebellar cortex. Structural and topographic characteristics of Lugaro cells, as well as the peculiarities of their contacts with the other cells of cerebellar cortex, in combination with the data on their neurotransmitter content, indicate that these cells play the role of inhibitory interneurons. PMID- 12891779 TI - [Computer morphometric analysis of dendrites of the type II Dogiel neurons]. AB - Dendrites of type II Dogiel neurons (the local receptors) stained with silver nitrate impregnation by Bilschovsky-Gross method, were analyzed with computer morphometry in cat large intestine. The following parameters have been measured: an area of dendritic receptor field, total area of all receptor elements and total length of all receptor branches. Significant individual variability was noted in the values of these parameters: the maximum value exceeded the minimum one by a factor of 41, 29 and 45, respectively. No correlation between these three parameters has been found. It is suggested that the morphological diversity of dendrites found in this study may serve as a basis of the functional variability of type II Dogiel neurons. PMID- 12891780 TI - [Characteristics of the cell proliferation in the developing human retina]. AB - The retinal primordia were studied in human embryos at developmental weeks 5-8 to examine the expression of proliferation-associated proteins (Ki67 and PCNA), the number of nucleoli in the nuclei of neuroepithelial cells (in respect to their distance to the apical surface) and the distribution of DNA-synthesizing cells after a short-term (20 min) incubation in vitro in serum-free medium containing BrdU. In 5-week embryos, retinal primordium contains the neuroepithelium with a typical structure. BrdU-positive nuclei and nuclei with small number of nucleoli were found in the basal portion of the ventricular zone. However, this organization was disturbed in the initial period of the formation of inner nuclear layer (week 6). At this stage DNA-synthesizing cells were seen even at the apical surface. In retinal primordia of embryos at weeks 6-7 there appeared an additional area of cell proliferation in the Chievitz layer and in the inner nuclear layer. In embryos at week 8 the dividing cells were concentrated in the outer nuclear layer, which has regained the organization, typical for a neuroepithelium. PMID- 12891781 TI - [Distribution of the labeled lipoproteins in the myocardium during perfusion of the isolated rat heart and adrenaline administration]. AB - The distribution of different classes of colloidal gold-labeled lipoproteins (LP) was studied at the ultrastructural level in rat myocardium using the isolated heart perfusion following the adrenaline stimulation. The distribution of label was examined in endotheliocytes, macrophages and in the interstitial space. The addition of adrenaline to Krebs solution during 30 min of heart perfusion in vitro was shown to increase the amount, but to decrease the velocity of penetration of labeled LP of very low and low density into the myocardium. The intensive utilization of lipid component from LP in the conditions of stimulation provides for the enhanced metabolic requirements of the cardiomyocytes and good preservation of their ultrastructure. It is possible that colloidal gold associated protein component of LP is incorporated by the endothelium more slowly. The label penetrates into the interstitium and is accumulated in the activated macrophages. If high density LP are added together with adrenaline into the perfusion medium the label does not leave the capillary wall, while macrophages and cardiomyocytes acquire the ultrastructural features of high functional activity. PMID- 12891782 TI - [Structural and functional changes in the histohematic barrier of the rat myocardium in the postischemic period]. AB - The aim of this study was to examine the myocardial histohematic barriers during the rehabilitation period after a cardiac arrest for 10 min. The electron histochemical analysis was applied to evaluate the permeability of histohematic barriers, to assess their ultrastructure and calcium localization. During 24 hours post resuscitation the destructive and compensatory-adaptive changes were found in the microcirculatory bed, accompanied by a pericapillary space edema, selective elevation of sarcolemmal permeability and calcium redistribution in cardiomyocytes. The structural and functional disturbances of histohematic barriers following the total ischemia are important factors of myocardial reperfusion lesions and of cardiodepression. PMID- 12891783 TI - [Morphofunctional state of the blood microcirculatory bed in the injured gastrocnemius muscle in rats during pharmacological stimulation]. AB - The state of hemomicrocirculatory bed and of local peripheral circulation was studied after the operational injury of rat gastrochnemius muscle without pharmacologic treatment and following a combined administration of alpha tocopherol and methyluracil. Local circulation in the regenerate of the treated animals was shown to increase not only as a result of increase of capillary density, but also due to the elevation in the number of functioning capillaries and veins. PMID- 12891784 TI - [C-cells of the thyroid gland in experimental osteoporosis]. AB - The population of thyroid C-cells of female rabbits with ovariectomy-induced osteoporosis was studied using immunohistochemical method with antibodies against calcitonin (CT) and morphometrically. The development of osteoporosis was confirmed using radiology and densitometry. 8 weeks after the operation, after the detection of hypercalcemia and hypercalciuria, some of the animals were given a correcting treatment with calcium-containing drug and vitamin D3 or with a combination of calcium drug with hormonal estrogen-containing drug. 4 weeks after the treatment was started, thyroid C-cells in animals that received the drugs, were significantly larger than in the cells of control and ovariectomized rabbits that received no treatment. Following 12 and 22 weeks, in the animals treated with hormonal drug, both calcium concentrations in blood and urine and C-cell dimensions were normalized. Large C-cells with an intensive reaction to CT were observed at weeks 12 and 22 (weeks 20 and 30 after the operation) in the thyroid gland of the animals that received no treatment and of the animals that received calcium with food. In the last case, the morphological changes of C-cells were more pronounced, despite the insignificant differences in blood calcium concentrations in both groups. These results permit to suggest that the intensity of CT synthesis by C-cells may be influenced by calcium concentrations not only in the blood, but also in the intestinal lumen. PMID- 12891785 TI - [Endocrine cell of the cow colonic mucosa]. AB - The epithelium of colon mucosa of the adult cow was studied using the methods of light and electron microscopy to demonstrate the endocrine cells. Individual fluctuations of the endocrine cell content was noted as well as an increase in the total number of endocrine cells in the rectal epithelium, that was more pronounced tan in other vertebrates. Four types of endocrine cells were demonstrated: EC, L, D, D1. The variability of dimensions and shape of granules in EC-cells was found to be less tan in other vertebrate species. In some D1 cells the mucous granules were observed along with endocrine ones, thus indicating the presence of "mixed" exo-endocrine cells. Undifferentiated endocrine cells were also detected that were found at the bottom of the crypts. The features indicated above are the specific peculiarities of endocrine apparatus of the mucosal epithelium of cow colon. PMID- 12891786 TI - [Morphometric characteristics of the kidney nephrogenic zone in newborn rats]. AB - The aim of this work was to give a quantitative characteristic of nephronogenesis and to select the morphometric criteria, which adequately reflect the dynamics of morphogenetic processes. As the study object, the nephrogenic zone of a newborn rat kidney was used, that was examined in sections stained using histological and histochemical methods. The development of nephron structures and of the stroma was followed, and it was established that grading the degree of differentiation of the structures, comparison the nephrogenic zone, reflects the system of current concepts on the inter-inductive influences, which are realized between the distal portions of collecting ducts and nephrogenic cells, epitheliocytes of developing tubules and stromal cells. The morphometric parameters obtained could be used for interpretation of the data on the velocity of nephron formation and tubulogenesis, as well as on the intensity and quality of vascular glomeruli formation. PMID- 12891787 TI - [The relief of the mineralization front of the natural surfaces and characteristics of the structure of the lacunar-canalicular system in the human mandible osseous tissue]. AB - Using the material from 12 corpses of males aged 19-44 years, the structural elements of mandibular osseous tissue were studied with scanning electron microscopy. The mosaic pattern was established in different areas which characterized the functional activity of particular region. In the bone surface, the resorbing, forming and formed (or inactive) areas were detected. Nutrition of the osseous tissue is provided by the lacunar-canalicular system, which is characterized by permanent remodelling of the bed. The development of osteocytic lacunae included the sequential formation of their bottom, walls and roof. These findings should be considered during preparation, conservation and selection of plastic material; they could also serve as a basis for theoretical and practical substantiation of the methods for optimization of reparative osteogenesis in the development of the new ways of directed influence on the processes of bone formation and resorption. PMID- 12891788 TI - [Correlation between the dimensions of the human paranasal sinuses and the skull type]. AB - To detect the dependence of the dimensions of maxillar and frontal paranasal sinuses on the age, sex, and the skull type in population inhabiting Karelya, 1566 maxillar sinuses and 1404 frontal sinuses were studied in radiographs of practically healthy subjects aged 3-85 years. The correlation between age, sex, dimensions of maxillar and frontal sinuses, and the skull type was established that is revealed in that the latitudinal dimensions of the sinuses prevail in brachiomorphous skull type, while the altitude dimensions are prevalent in dolichomorphous type regardless the sex and age. Total dimensions of maxillar sinuses are larger in dolichomorphous skull type: in males that is especially pronounced for maxillar sinuses, while in females--for frontal sinuses. PMID- 12891789 TI - [Unmyelinated nerve fibers]. AB - The paper presents a critical review of various current concepts of the structure and kinetics of unmyelinated nerve fiber. A classification of nerve fibers, different from the earlier ones, is proposed, that demonstrates not only the morphological fiber types, but also the kinetics of their reversible transition stages from non-glial to myelinated fiber. Evidence is presented to show the erroneousness of conceptions, still appearing in many publications, that consider the unmyelinated nerve fiber as "the Remak's cable type fiber". According to the current data, "Remak's fiber" is a glial-neurite complex, i.e. a bundle of unmyelinated nerve fibers covered with a single glial cell. Using the electron microscope, it was demonstrated that comparable glial-neurite complexes of myelinated nerve fibers, formed in CNS in a similar way by a single oligodendrocyte, cannot be named a single fiber. Cutting the nerves makes visible that the single fibers forming "the Remak's fiber" stem from different cells, therefore they cannot be a single "fiber". It has been shown for the first time experimentally, that in extreme situations, as a result of contraction of gliocyte processes, unmyelinated fibers may "leave" the glial-neurite complexes and become the nonglial fibers. Some data are presented that may serve as criteria for differentiation of unmyelinated fiber with a stratified sheath from developing myelinated fiber. PMID- 12891790 TI - [The histaminergic brain neuron system]. AB - This review describes the current concepts of distribution, morphological peculiarities and functions of histaminergic structures in human and animal brain. In adult mammals and humans, the perikarya of histaminergic neurons are localized exclusively in the hypothalamus, mainly in its tuberomamillary nuclei, where 5 subgroups are distinguished. It is only there that the syntheses of neuronal histamine takes place and histamine-synthesizing enzyme histidine decarboxylase and corresponding mRNA were found. The processes of histaminergic neurons innervate practically all the brain regions. Histamine action is mediated by three types of receptors: H1, H2 (postsynaptic) and H3 (presynaptic), which are widely distributed in the brain. Histaminergic brain system is involved in the regulation of various functions, systems and reactions of the organism and participates in the pathogeneses of many disturbances and diseases. The review provides the detailed description of the localization of histaminergic neuron perikarya in mammals, the existing classifications of histaminergic neurons of rat brain, the distribution of histaminergic fibers in various brain regions, the organization of human histaminergic brain system, histaminergic neuron ultrastructure, afferent innervation and regulation. PMID- 12891791 TI - Ready for the transactions rule? Get started with code sets. PMID- 12891792 TI - Coders today: where they work, what they earn. PMID- 12891793 TI - The power of persuasion. Proven strategies inspire physicians to improve documentation. PMID- 12891794 TI - What happened to PEPP? QIOs plan for hospital payment monitoring program. PMID- 12891795 TI - AHIMA project offers insights into SNOMED, ICD-9-CM mapping process. PMID- 12891796 TI - The 1970s: "we must be able to talk both languages". PMID- 12891797 TI - Preparing for suspension of disclosure accounting. PMID- 12891798 TI - Saving time: a physician's perspective. PMID- 12891799 TI - Stealth software deters PHI theft. PMID- 12891800 TI - Practice brief. Managing and improving data quality (updated). PMID- 12891801 TI - OIG ambulance guidance applies to all healthcare sectors. PMID- 12891802 TI - International refined DRGs globalize coding. PMID- 12891803 TI - SARS tops healthcare concerns. Coding role vital to reporting deadly disease. PMID- 12891804 TI - Deeply affected. As the nation ages, Alzheimer's will strike more people close to us. PMID- 12891805 TI - New horizons. Early diagnosis of Alzheimer's means new implications for care. PMID- 12891806 TI - 'Sea-ing' results. An aquarium can help boost Alzheimer's patients' appetites. PMID- 12891807 TI - How 'sweet' it is. A holistic approach to Alzheimer's nurtures patient and caregiver. PMID- 12891808 TI - Making connections. Silverado builds relationships with residents, relatives and staff. PMID- 12891809 TI - New age care. It's time to focus on the need-driven, market niche of Alzheimer's and dementia specialty care. PMID- 12891810 TI - [Cancer detection: do not close the debate...]. PMID- 12891811 TI - [Breast cancer detection and medicalization of public health]. AB - The aim of reducing breast cancer mortality announced in the report on health in France published in 1994 strongly implicated the promotion of screening and early surgical treatment. However, this mortality has not evolved significantly in France, although more and more cancers have been found and operated upon. The increased prevalence rates of histological diagnoses of breast cancer obtained from the studies of autopsies published has shown that the screening has discovered non-terminal, benign cancers. It has been estimated that 120 of the surgical operations conducted per day in France in 1999 have been carried out to remove non-terminal, benign cancers. Public health has an ethical responsibility to face in terms of the allocation of resources and must remain vigilant in order to avoid excessive screening and unnecessary interventions. PMID- 12891812 TI - [Medical-social study of women incarcerated in the Fleury-Merogis prison in 2000 and 2001]. AB - This socio-medical study was conducted in the women's penitentiary of Fleury Merogis in 2000 and 2001 and includes a total of about 500 patients of whom 70% are between the ages of 18 and 39. Their level of education varies; 63% of them are foreigners, of which 60% do not speak any French. Their physical state of health is satisfactory in general, but more than half of them suffer from minor psychological problems often secondary to the prison's conditions. Major psychiatric illness is rare. Among their addictions, the one noted most frequently is smoking; alcoholism and drug addiction are less widespread. The author underlines the need for a comprehensive care structure for the prisoners, on the socio-economic and family levels as well as the physical and psycho pathological levels. He advocates in order that the medical care become the activity of a single service of Internal Medicine, and no longer as it exists in the present system in France with two distinctly separate departments--one in charge of caring for the body, and the other in charge of caring for the mind. Finally, he emphasises the need for a true sense of team work among all of the actors involved in the care of these women in difficult situations. PMID- 12891813 TI - [Towards a daily social ethic]. PMID- 12891814 TI - [Health in schools or healthy schools?]. AB - Comparing the existing documentation on the European Network of Health Promoting Schools (ENHPS, in French: REES) and on the French School Health Service, many differences as well as similarities are appearing. In Switzerland, the ENHPS/REES is presently well developed. Facing new attitudinal and behavorial problems in schools, a recent experience in Geneva (Switzerland) includes health promotion as one of its objectives. PMID- 12891815 TI - [Today's youth who are giving us a scare...]. PMID- 12891816 TI - [Morbidity diagnosed in the general medicine public structures in Tunisia]. AB - The study of morbidity in general medicine is very useful in order to adapt training curricula to the reality of medical practice in the first degree. The objective of this work was to describe the morbidity charted in general medicine in Tunisia's Sousse region. It consisted of a prospective and descriptive study involving six basic health centres in Sousse and was conducted over the course of 30 randomly selected days during the year 2000. There were 4022 consultations included in the study which were described according to the SOAP plan. The coding of the charted diagnoses was carried out according to the International Classification of Primary Care (CISP). There were 98% of the consultants who were self-referred and 84% of the consultations corresponded to new cases. The consultants' sex-ratio was 0.5 in favour of females with an average age of 27 years. There were 4,597 diagnoses noted utilising 336 codes from the CISP classification system. The "Respiratory" chapter took the first place on the list (39.6%) followed by the "Circulatory" chapter. The "Top 30" list of the most often charted diagnoses totaled 72% of the consultations among which the three most frequent health problems were pharyngitis (14.4%), acute bronchitis (8.3%) and arterial hypertension (7.4%). This study demonstrates that the practice of general medicine is characterised by instinctive first solutions and versatility. The predominance of cardiovascular and respiratory illnesses illustrates the double burden of morbidity which is faced by Tunisian general medicine practitioners. The "Top 30" list should serve as basis for planning pregraduate and continuing medical training. PMID- 12891817 TI - [Research in general medicine, role of the medical thesis]. AB - General practice doctors are naturally presented as being actors on the front lines of public health. The ability of general practitioners to be aware of and deal with public health questions largely depends upon their training. Perhaps one could consider the general practitioner's thesis as a preparatory analytical work within the area of their future field of practice. Do these theses serve to provide food for thought on general practice and its contribution to public health, and if not, could they? An analysis conducted within four medical schools in western France demonstrates that the work produced for general medicine identified as such only constitutes a fraction of the overall number of medical (non-university) theses (approximately 5%). Two possible explanations may be put forward. First, on the one hand, the theses highly depend on the context of the training and the work produced is a direct result of this. Second, on the other hand, the current methods of indexing notes in the university databases does not enable general medicine to be clearly seen and visibly recognised as an academic discipline. Two questions then remain: Can medical theses be considered as research? Should research in general medicine be carried out solely by general practitioners? It is vital that resources from outside the medical field be raised and mobilised for general practice research, whose themes are multi disciplinary and not only clinical. PMID- 12891818 TI - [Quality of care in psychiatric hospitals: literature review and perspectives]. AB - Within hospitals, a plan for quality improvement consists of numerous steps to be taken by an inclusive management team, implicating all of the actors involved. The establishment of an instrument panel of indicators is the outcome of such a participatory process. The existence of best practice referentials allows for the construction of a coherent plan to improve the quality of care. A query of the francophone bibliographic databases reveals very few publications from internal audits on the relevance of care. Nevertheless, external audits demonstrate the significance of evaluation of the quality of care in psychiatric hospitals. The authors are therefore surprised to note the existing hiatus between the wealth of references and their lack of being used in the publications of the professionals concerned. The analysis of the relevance of the medical decision is not widely accepted. Clinical practice is still considered as an art form even though it has become a technique which now calls on the collective experience of peers. PMID- 12891819 TI - [Sexually transmitted diseases, AIDS and prostitution: a case study in Guinea Conakry]. AB - In Africa, many public health interventions related to the fight against HIV/AIDS are aimed at women commercial sex workers. The practices of sexual labour and prostitution are not universal, and considering them within their specific cultural context is vital to understanding these women's needs and the prevention of HIV/AIDS. An exploratory qualitative study inspired by well-rooted theory was conducted with 14 women working within formally renowned prostitution sites in Guinea-Conakry. It aimed at identifying the context and general preoccupations of these women, within which are embedded sanitary concerns. The study showed that their sexual behaviours are mostly influenced by collective, relational and economic aspects which are in turn evaluated by the women within the framework of their continuous and changing life cycle. PMID- 12891821 TI - Food-consumption patterns in central West Africa, 1961 to 2000, and challenges to combating malnutrition. AB - We discuss food-consumption patterns in Central West Africa from 1961 to 2000 and some implications for combating malnutrition. The availability of food in the region improved in the 1960s, declined sharply in the 1970s and the early 1980s, and has shown a modest positive trend since the mid-1980s. Notwithstanding obvious progress over the past decades and in the region as a whole, food availability today remains below the required levels for large parts of the population and appears unstable over time, particularly in the Sahelian zone. On average, diets in this zone contain fewer than 2,200 kcal, compared with almost 2,500 kcal in the coastal zone. Conversely, protein deficiency is more common in the coastal zone, where a typical diet contains only 45 g of protein, compared with 60 g in the Sahelian zone. Furthermore, consumption is showing a dietary shift toward cereals, while yield growth lags far behind population growth. The associated import dependency and pressure on land seem to gain significance regardless of the region's agro-ecological capacity to increase and to substitute cereal imports for locally produced food. Moreover, food consumption appears responsive to income changes (calorie-income elasticity ranges from 0.25 to 0.62), while, in turn, it has a significant impact on nutritional outcomes (stunting-calorie elasticity of -1.42). We conclude that combating malnutrition requires first broad-spectrum income growth, and next specific policies that promote the yield and the contribution to diets of nutritious food produced within the region. PMID- 12891820 TI - Malnutrition and dietary protein: evidence from China and from international comparisons. AB - Malnutrition, as measured by anthropometric status, is a powerful risk factor for illness and elevated death rates throughout life. Understanding the relative importance of disease, dietary quantity, and dietary quality in causing malnutrition is therefore of major importance in the design of public policy. This paper contributes to the understanding of the relative importance of quantity and quality of diet by utilizing aggregate data to complement previously reported individual-level studies. Three compilations of anthropometric data--one involving subjects from 13 provinces in China, another involving subjects from 64 counties in China, and a third involving 41 populations in 40 countries--are used to examine the relative importance for human growth of inadequacies of dietary energy and protein. The analysis involves regressing average adult heights and weights against estimates of average energy and protein availability (by province, county, or country) and per capita incomes. We use protein availability in part as a marker for overall quality of the diet, while recognizing that protein is far from perfectly correlated with dietary fat or micronutrient availability. The paper discusses issues of both data quality and statistical methodology, and points to relevant resulting caveats to our conclusions. Subject to these limitations, all three analyses suggest that, at the levels of dietary intake in these populations, lower protein intake is related to growth failure whereas lower levels of energy availability are not. The protein effect appears stronger for males than for females. PMID- 12891822 TI - An evaluation of the national food and nutrition policy of Bangladesh. AB - Food and nutrition policy activities directed toward improvement of the nutritional status of the people of Bangladesh began in the 1980s. The government formulated a national food and nutrition policy and approved it in 1997. Qualitative methods, including observational techniques, in-depth interviews of the key informants, and focus group discussions, were used to collect information on the strengths, weaknesses, opportunities, and threats (SWOT) of the policy. The information obtained has been transcribed and analyzed using this model. The strengths of the policy are that it is a consensus document that emphasizes human rights, was formulated by a multisectoral approach, complements other government policies, and has broad goals and wide-ranging objectives. The weaknesses include lack of implementation, monitoring, and evaluation guidelines; lack of strong government commitment; inadequate support of policy makers; perhaps an excessively ambitious target; and ignorance of past lessons learned. The opportunities include the scope of social mobilization, the wide scope of the policy, suggested programs and measures to improve nutritional status, a congenial policy environment, and the ability to modify the scope of the policy as needed. The threats to the policy are lack of knowledge of the policy, lack of resources to implement the policy, tension between technical people and bureaucrats, vested business interests, and, possibly, discontinuity of political commitment. The key to reducing the weaknesses of the food and nutrition policy of Bangladesh and minimizing the threats to it is for the stakeholders in the policy to coordinate efforts to use the strengths and opportunities of the policy to effectively implement it. PMID- 12891823 TI - Timed activity studies for assessing the energy expenditure of women from an urban slum in south India. AB - Time-disposition studies are necessary for computing energy requirements of populations. This study captures the rich information on the timed activity pattern of adult women from poor households engaged in home-based work. We studied 34 women beedimakers (cigarette makers), 21 tailors, and 34 homemakers. Data were collected by direct observation of the women's activities on a typical day. Time spent on related activities was pooled and classified as sleep, household work, child care, occupational work, and residual work. These were further categorized on the basis of our published work on the energy cost of women's activities and the World Health Organization (WHO) classification of occupational activities as sedentary, moderate, and heavy. Most of the household activities could be classified as moderate to heavy (> 2.2 times basal metabolic rate [BMR]). Childcare activities were distributed on a scale from sedentary to heavy, whereas occupational activities, such as beedimaking and tailoring, were sedentary (< 2.2 BMR). Homemakers spent significantly more time on moderate to heavy work (p < .05) than beedimakers and tailors. Women working for income spent only four to six hours on occupational work, which was possible because they reduced the time spent on heavy work (i.e., housework), and reduced the time on personal care. Still, more than 80% of women could not put in eight hours of paid work. Thus, women in the home-based sector constantly negotiate among time spent on heavy household work, child care, and occupational work in order to continue in the labor market. PMID- 12891824 TI - Combating iodine and iron deficiencies through the double fortification of fish sauce, mixed fish sauce, and salt brine. AB - Two iodine and seven iron compounds were tested for use in the fortification of pure fish sauce, mixed fish sauce, and salt brine for cooking as a means to combat iodine and iron deficiencies. Ferrous sulfate, sodium iron ethylenediaminetetraacetic acid, ferric ammonium citrate, and ferrous lactate were combined with potassium iodide with no effect on sensory quality. Product shelf-life testing revealed that no iron or iodine losses occurred during a three month storage period. Although the color of most products darkened, the color was not significantly different from that of nonfortified products after two to three months. Sensory home-use tests revealed that the fortified products were acceptable to highly acceptable, with only 1.2% to 8.2% of the dishes cooked using the fortified products being reported as discolored. The cost of fortification was minimal, at 0.13 to 2.73 baht per bottle (750 ml) (42 baht = US$1). Consequently, these products show a potential for inclusion in national programs for the prevention of micronutrient deficiencies in Asian countries where fish sauce and its products are routinely consumed. PMID- 12891825 TI - Red palm oil supplementation: a feasible diet-based approach to improve the vitamin A status of pregnant women and their infants. AB - This double-blinded, randomized, controlled study was designed to study the effect of dietary supplementation with red palm oil during pregnancy on maternal and neonatal vitamin A status. A total of 170 women were recruited at 16 to 24 weeks of gestation and randomly assigned to an experimental group that received red palm oil to supply approximately one recommended dietary amount (RDA) (2,400 micrograms) of beta-carotene or to a control group that received an equivalent volume of groundnut oil. The women received the oils for a period of 8 weeks, starting at 26 to 28 weeks of gestation and extending to 34 to 36 weeks of gestation. The mean postintervention (34 to 36 weeks) levels of serum retinol were 1.20 +/- 0.22 (SD) mumol/L (95% CI, 1.15-1.25) in women receiving red palm oil and 0.73 +/- 0.15 mumol/L (95% CI, 0.69-0.77) in their infants; these levels were significantly higher than those in women receiving groundnut oil (1.07 +/- 0.26 mumol/L; 95% CI, 1.01-1.13; p < .01) and their infants (0.62 +/- 0.17 mumol/L; 95% CI, 0.57-0.67; p < .001). A significantly lower proportion of women in the red palm oil group than in the control group had vitamin A deficiency (serum retinol levels < 0.7 mumol/L) after intervention (1.5% vs. 9.7%). The proportion of women having anemia was significantly lower (p < .01) in the red palm oil-supplemented group (80.6%) than in the control group (96.7%). The mean birthweight and gestational age of the infants did not differ significantly between the two groups. An increased risk of low birth-weight (p = .003) and preterm delivery (p = .000) was observed with decreasing serum retinol levels in the third trimester of pregnancy. These results show that red palm oil supplementation significantly improved maternal and neonatal vitamin A status and reduced the prevalence of maternal anemia. Maternal vitamin A status in the later part of pregnancy is significantly associated with fetal growth and maturation. Hence red palm oil, a rich source of bioavailable vitamin A, could be used as a diet-based approach for improving vitamin A status in pregnancy. PMID- 12891826 TI - A multinutrient package of iron, vitamin A, and iodine improved the productivity and earnings of women tea pickers in south India. AB - Tea-picking is a highly skilled activity that is usually performed by women. This study, conducted on the Balanoor Plantations, India, from 1996 to 1998, was successful in empowering 339 women pickers and their families to take iron (60 mg of elemental iron two times a week) and vitamin A (1,600 IU) once a week, and to purchase subsidized iodized salt (30 ppm) from the plantation ration shop. The average hemoglobin level of the pickers rose significantly (p < .001) from 11.0 g/dl at baseline to 11.9 g/dl at the end of the nine months of intervention. The average amount of tea per picker increased significantly (p < .001) from 22.9 to 25.6 kg. There was a significant decrease in the number of "moderate pickers," who picked between 14 and 25 kg per day, and a significant increase in the number of "good pickers," who picked more than 25 kg per day, and the earnings of the majority of the pickers increased. The management of the estate where the intervention occurred benefited from a decrease in the number of pickers needed during the supplement period from 2,857 to 2,763, with no significant change in the yield per hectare in the two years. The yield per hectare on the control estate was not significantly different from that on the intervention estate, and the average amount of tea picked per worker was the same for the two periods (20.8 and 20.7 kg). PMID- 12891827 TI - A method of preserving and testing the acceptability of gac fruit oil, a good source of beta-carotene and essential fatty acids. AB - Gac fruit (Momordica cochinchinensis Spreng) is indigenous to Vietnam and other countries in Southeast Asia. Its seed pulp contains high concentrations of carotenoids, especially the provitamin A, beta-carotene. In northern Vietnam, gac fruits are seasonal and are mainly used in making a rice dish called xoi gac. The purpose of this study was to develop a method to collect and preserve gac fruit oil, to evaluate the nutritional composition of the oil, and to assess the acceptability of the gac oil by typical Vietnamese homemakers. One hundred women participated in training to learn how to prepare the fruits and operate the oil press. The women also participated in a survey of gac fruit use and their habitual use of animal fat and vegetable oil. Among all the participants in the training and surveys, 35 women actually produced oil from gac fruits grown in the village, using manual oil presses and locally available materials. The total carotene concentration in gac fruit oil was 5,700 micrograms/ml. The concentration of beta-carotene was 2,710 micrograms/ml. Sixty-nine percent of total fat was unsaturated, and 35% of that was polyunsaturated. The average daily consumption of gac fruit oil was estimated at 2 ml per person. The daily beta carotene intake (from gac fruit oil) averaged approximately 5 mg per person. It was found that gac oil can be produced locally by village women using manual presses and locally available materials. The oil is a rich source of beta carotene, vitamin E, and essential fatty acids. Although the beta-carotene concentration declines with time without a preservative or proper storage, it was still high after three months. The oil was readily accepted by the women and their children, and consumption of the oil increased the intake of beta-carotene and reduced the intake of lard. PMID- 12891828 TI - Is dietary diversity an indicator of food security or dietary quality? A review of measurement issues and research needs. PMID- 12891829 TI - Response to the FNB Supplement on the Positive Deviance Approach to Improve Health Outcomes. PMID- 12891830 TI - [Prevention of bacteremia from central venous catheters in hemodialysis: value of application of a mixture of rifampicin and protamine at the insertion site]. AB - During the last 8.5 years, authors observed only 3 bacteremias out of 249 catheters in place in different sites corresponding to a total of 10,063 days with central venous catheterization in hemodialysis (0.30/1000 catheter days). This good result seems to be correlated mainly with the preventive application, on catheter insertion-site, of a rifampin and protamine sulphate mixture. This protocol appears worthy to be known because, at the present time, whatever the new preventive strategies, infection rates reported in the literature are generally more consequent and it is necessary to consider all the possibilities of decreasing them. PMID- 12891831 TI - [Hantavirus infections in Ile-de-France]. AB - Hantavirus infections, Puumala serotype, is a well-known disease in the northeast of France, but not in the Paris area, despite regularly diagnosed cases. A retrospective study was performed from January 1999 to December 2000 to assess the clinical and epidemiological characteristics of this disease in the "Region Ile-de-France". Fourteen cases were diagnosed. All required hospitalisation. Patients presented usually with high fever, pain, renal failure, moderate thrombocytopenia and, sometimes, transitional acute myopia witch is pathognomonic of the disease. In each case, a contact with a forest was found. Twelve patients recovered completely. One patient with pre-existing chronic renal and hepatic failure died and another developed a persistent arterial hypertension. Invasive procedures were often used before the diagnosis. Hantavirus infections does exist in the "Region Ile-de-France". Failure to recognise this disease in this area lead to unnecessary invasive procedures and hospitalizations. PMID- 12891832 TI - [Retroperitoneal fibrosis, an unrecognized inflammatory disease. Clinical observations and review of the literature]. AB - Retroperitoneal fibrosis (RF) is a rare disease, typically with an insidious clinical course. The peak incidence is seen in patients 40 to 60 years of age and mostly in man. The characteristic finding in this disease is a periaortic fibrous mass that often surrounds the ureters. Although usually regarded as an obstructive uropathy, there has been growing recognition of the condition as a generalized disease. It may have a wide variety of manifestations including mediastinitis, thyroiditis and sclerosing cholangitis. The most common mode of presentation remains abdominal or flank pain with uremia, anemia and a high sedimentation rate. Although ultrasound and renal scintigraphy may contribute to the general evaluation of patients with RF, CT-scanner is the preferred imaging method. The multiplanar imaging capability of magnetic resonance may facilitate assessment of disease extent. The pathogenesis of the disease remains unknown. Steroids and, more recently tamoxifen, appear to be effective in the treatment of the RF. In most instances, RF does not lead to long-term morbidity or affect survival. The three cases of RF reported herein illustrate the varied mode of presentation and the response to the treatment. PMID- 12891833 TI - [Acute renal insufficiency secondary to interferon]. AB - Alpha interferon is increasingly used in the treatment of malignancies and viral hepatitis. Renal involvement after its use consist in more cases on proteinuria. We report one case of acute renal failure without nephrotic syndrome but with tubulointerstitiel infiltration by mononuclear cells in 65 old man who had before interferon therapy 151 mumol/l of serum creatinine. Interferon was administrated for digestive carcinoid neoplasia and bone and liver metastasis. Outcome was improvement of renal function after withdrawn of interferon, dialysis and steroid treatment. PMID- 12891834 TI - Not "just a nurse". PMID- 12891835 TI - Current controversies in the understanding of necrotizing enterocolitis. Part 1. AB - Necrotizing enterocolitis (NEC) has widespread implications for neonates. While mostly affecting preterm neonates, full-term neonates, especially those with congenital heart disease, are also at risk. Although the exact pathogenesis of NEC remains elusive, three major factors, a pathogenic organism, enteral feedings, and bowel compromise, coalesce in at-risk neonates to produce bowel injury. Initiation of the inflammatory cascade likely serves as a common pathway for the disorder. Clinical signs and symptoms range from mild feeding intolerance with abdominal distension to catastrophic disease with bowel perforation, peritonitis, and cardiovascular collapse. Vigilant assessment of at-risk neonates is crucial. When conservative medical management fails to halt injury, surgical intervention is often needed. Strategies to decrease the incidence and ultimately prevent NEC loom on the horizon, such as exclusive use of human breastmilk for enteral feedings and administration of probiotics. PMID- 12891836 TI - A case series of five infants with scimitar syndrome. AB - Scimitar syndrome is a rare association of congenital cardiopulmonary anomalies consisting of a partial anomalous pulmonary venous connection of the right lung to the inferior vena cava, right lung hypoplasia, dextroposition of the heart, and anomalous systemic arterial supply to the right lung. To date, this syndrome has not been reported or discussed in the nursing literature. In this article, 5 patients are presented to illustrate the variable presentation of this syndrome, ranging from infants who are asymptomatic to those with heart failure and severe pulmonary hypertension. The clinical signs and symptoms, diagnostic dilemmas, and medical and surgical management are discussed. The triad of respiratory distress, right lung hypoplasia, and dextroposition of the heart should alert the clinician to the possibility of this syndrome. PMID- 12891837 TI - A critical analysis of the routine testing of newborn stools for occult blood and reducing substances. AB - Stool tests for occult blood or reducing substances were introduced in the neonatal intensive care unit (NICU) as potential aids in the early recognition of necrotizing enterocolitis (NEC) in high-risk neonates, and have been recommended by some as routine nursing procedures. Neither the performance characteristics of these tests with respect to NEC, nor their indirect impact, were evaluated formally before widespread adoption into clinical care. The published evidence suggests that these tests are not useful as diagnostic or screening tools. There is no evidence that routine stool screening for occult blood or reducing substances predicts NEC or decreases the rate or severity of this disease. The direct costs of the tests are significant. A greater concern is their potential unintended consequences, which include the cost of secondary tests, restricted nutritional intake, and the accumulation of distracting, useless data. The logistics of maintaining quality control, the demands on nursing time, and the cost of testing are increasingly important considerations. This installment of Focus on the Physical diverges from a step-by-step systematic physical assessment by addressing the utility of testing neonatal stools for occult blood and reducing substances as aids in the early diagnosis or prevention of NEC. Using the information from these tests requires a framework for understanding their rationale, the test performance characteristics in the NICU setting, and the potential benefits, costs, and risks of their routine use. PMID- 12891838 TI - A descriptive study of bottle-feeding opportunities in preterm infants. AB - PURPOSE: The purposes of this study were to describe patterns of bottle feeding opportunities in preterm infants and to analyze the relationships between feeding opportunities and outcomes. SUBJECTS: The smaple was composed of 25 preterm infants who were bottle fed. DESIGN: Retrospective, correlational. METHODS: Data were collected from medical records and analyzed descriptively. Correlation coefficients among variables were computed. MAIN OUTCOME MEASURES: Postconceptional age (PCA) at the time of full bottle feedings and transition time from the first to full bottle feedings. PRINCIPAL RESULTS: The number of bottle feedings received per day during the transition from first to full bottle feedings was inversely related to length of transition to full bottle feedings. There was no relationship between the number of bottle-feeding opportunities and PCA at full bottle feedings. CONCLUSIONS: The achievement of full bottle feedings may be facilitated by increased bottle-feeding opportunities. More research about the relationships among feeding readiness, outcomes, and opportunities is needed. PMID- 12891839 TI - Parent education after newborn congenital heart surgery. AB - The moment an infant is diagnosed with a congenital heart defect, parents experience a mixture of shock, disbelief, fear, anger, and often a profound sense of sadness. In the midst of these emotions they must learn to provide for the special needs of their infant. Providing parents with the knowledge and skills to care for their infant during this stressful time requires the concerted effort of a multidisciplinary team who can provide clear, concise, and consistent communication. This article provides a step-by-step guide to the information needs of families who are preparing to take their infants home after cardiac surgery. Parents must understand the underlying cardiac defect and the goals and objectives of the surgical repair. In addition to normal newborn care, parents need to learn about care of the incision, nutritional support, and how to safely administer prescribed medications. Potential complications and when to call their health care provider or seek emergency care are an important focus of teaching. Information about infant development, challenges specific to their infant, and pragmatic strategies to support normal development are of prime interest to parents. Written reference materials, a Web resource list, and a family discharge teaching tool provide tangible resources for the family after discharge. PMID- 12891840 TI - Caring for your baby after heart surgery. PMID- 12891841 TI - [What is the cause of sunlight-induced rash?]. PMID- 12891842 TI - [Returning home sick from vacation. Harmless fever or malaria?]. PMID- 12891843 TI - [Interview with Prof. Dr. Erich Kroger, Dusseldorf. A question about travel should become an anamnesis standard]. PMID- 12891844 TI - [Specialty societies want to reform the embryo protection law. Producing, selecting, freezing in case of future need?]. PMID- 12891845 TI - [Confusing the broad-leaf garlic plant and meadow saffron]. PMID- 12891846 TI - [Functional food: special additives to make foods healthier. Margarine instead of statins?]. PMID- 12891847 TI - [More coordination and transparency in therapy of breast cancer. Every second woman requests breast centers]. PMID- 12891848 TI - [Value of radiotherapy in breast cancer. Breast preservation rises to 80%]. AB - The indication for radiotherapy for mamma carcinoma is determined in particular by the macroscopic and microscopic tumor findings, the number of lymph nodes identified, receptor status, the surgical report and (neo)adjuvant chemotherapy. Irradiation is applied in particular in conjunction with breast-preserving first line treatment in the case of ductal in situ carcinomas, and postoperatively following breast conservation in the case of invasive mamma carcinoma. The indication for irradiation following mastectomy is controversial and is determined by tumor size and location, together with further factors (see the Guidelines of the German Society of Senology). The indications applying to the irradiation of regional lymph nodes are determined by the sites of the respective metastases. The combination of radio- and chemotherapy is carried out in close cooperation between oncologists and radiologists. PMID- 12891849 TI - [Surgical therapy of breast carcinoma. Much of the fright is lost]. AB - Ever since breast cancer has become looked upon as a local and simultaneously a systemic disease, local treatment has become ever less radical. Today, some 70% of all mamma carcinomas can be treated with breast-preserving techniques. Following pre-operative downstaging by primary neoadjuvant chemotherapy, even the majority of large tumors can be managed with conservative surgery. With the exception of ductal carcinoma in situ (DCIS) axillary lymph node clearance is currently standard practice. For an assessment of nodal status, at least 10 lymph nodes must be obtained and examined histologically. The sentinel lymph node biopsy performed under study conditions should be left to experienced specialized centers. Postoperative irradiation of the preserved breast is an element of the overall concept of breast-preserving surgery. The spectrum of surgical options covers breast-preserving procedures, skin-preserving mastectomy and amputation, which cannot always be avoided. With regard to plastic reconstruction, a differentiation should be made between the possibilities for reconstruction during primary surgery and reconstruction at a later date. Since postoperative radiotherapy can considerably diminish the results of reconstruction, current tendency is towards secondary reconstruction. PMID- 12891850 TI - [Health care deficiencies in therapy of breast cancer. Are breast centers the solution?]. PMID- 12891851 TI - [Prevention of coronary heart disease with statins. Which patients profit?]. AB - Statins are generally considered as safe drugs with a very favorable cost efficacy-ratio. Calculation of health expenses limits the prescription of statins in primary prevention to persons on high risk (i.e. 20% risk of a coronary infarction within the next 10 years). Prescription of statins in secondary prevention of atherosclerosis is mandatory. Advanced age, impairment of renal function or polypharmacotherapy increase the incidence of severe side-effects, especially myopathy and rhabdomyolysis. These patients should be given statins with lesser risk of these side-effects. Combination of statins with fibrates has attracted public concern, but may be indicated in persons with otherwise not treatable hyperlipidemia. These patients need intensive monitoring, just as patients on other drugs that are metabolized via the enzyme CYP3A4. Patients on statins should get dietary counseling, as an appropriate diet increases the effect of statins. PMID- 12891852 TI - [Manifestations of organic disorders can often lead to diagnostic errors. Panic attack is often misdiagnosed]. PMID- 12891853 TI - [Acute pancreatitis? What is the cause?]. PMID- 12891854 TI - [First fusion inhibitor is available. No entry for HIV-1 viruses]. PMID- 12891855 TI - [Study examines prescriptions for hypertension. Physicians adhering to guidelines very well]. PMID- 12891856 TI - [Aggressive and early combined drug therapy. Antidiabetics for prevention of myocardial infarct]. PMID- 12891857 TI - [A new osteoporosis drug. Recombinant parathyroid hormone rebuilds bone]. PMID- 12891858 TI - [Concentration problems, delayed reaction. "Diagnosis at a glance" points to hepatic encephalopathy]. PMID- 12891859 TI - [Back aches often become a chronic problem. Sodium channel block against chronification]. PMID- 12891860 TI - [NMR shows how beta blockers change the failing ventricle. Decreased size and better output performance]. PMID- 12891861 TI - "Shaping" population health: a challenge for public health. PMID- 12891862 TI - My personal perspective on the obesity epidemic. PMID- 12891863 TI - Controlling an epidemic: the problem of overweight in children and adolescents. PMID- 12891864 TI - Obesity epidemic: we have to spend to save. PMID- 12891865 TI - [Increased obesity--decreased interest?]. PMID- 12891866 TI - Issues of causality in the history of occupational epidemiology. AB - Occupational epidemiology has its roots in classical medicine. However, it became a quantitative discipline only in the 20th century, through the pioneering work of individuals such as Case, Lloyd, and Selikoff and organizations such as the Division of Occupational Health of the U.S. Public Health Service. Studies of chemical dye workers, bituminous coal miners, smelting workers, and uranium miners have been especially important sources of innovations in methodology and in development of logical reasoning leading to acceptance of causal relationships of occupational exposures that lead to respiratory diseases and cancer. The cooperation of labor unions, such as those of steel and asbestos workers, has often been a crucial factor in providing essential data. PMID- 12891867 TI - Does overweight in infancy persist through the preschool years? An analysis of CDC Pediatric Nutrition Surveillance System data. AB - OBJECTIVE: To determine whether overweight in infancy (0-11 months) and young childhood (12-35 months) persists through the preschool years. METHODS: Analysis of longitudinal surveillance data for 380 518 low-income children monitored in the U.S. Pediatric Nutrition Surveillance System from birth to age 59 months. Overweight was defined as weight-for-height > or = 95th percentile. We determined the proportion of the children (overweight vs non-overweight) above or below the 95th percentile of weight-for-height at the later ages. RESULTS: The relative risk (RR) for overweight among overweight infants (vs non-overweight infants) at 1, 2, 3 and 4 years old was 4.3, 3.5, 3.3, and 2.9, respectively. 62.5% of overweight 3-year-old was still overweight a year later, but only 4.1% non overweight 3-year-old became overweight a year later (RR = 15.2). However, low birth weight children had the highest RR to remain overweight after they became overweight compared to normal and high birth weight children. CONCLUSIONS: Overweight during infancy persists through the preschool years. Tracking of overweight appears to become stronger as children get older and is more pronounced among low birth weight children than normal or high birth weight children. Monitoring preschoolers' height and weight status should be a strategy for preventing of obesity in adolescence and adulthood. PMID- 12891868 TI - Alarming trends in pediatric overweight in the United States. AB - OBJECTIVES: A comprehensive review of the pediatric overweight literature was undertaken to examine secular changes in the prevalence of pediatric overweight, the current magnitude and scope of pediatric overweight, and the identity of high risk groups in the U.S. METHODS: Articles published in the last two decades and studies of nationally representative numbers of children were highlighted. RESULTS: An unprecedented three-fold increase in the prevalence of pediatric overweight has occurred in recent decades in the U.S. and evidence suggests that this trend is continuing unabated. While no sociodemographic or racial group has escaped this trend, particularly at risk are African American girls, Hispanic girls and boys, and children from low-income house-holds. CONCLUSION: The rising prevalence of pediatric overweight in the U.S. is a harbinger of increases in diabetes, cardiovascular disease and numerous other health problems. Furthermore, disparities in pediatric overweight along ethnic and socio-economic lines are expected to further exacerbate current disparities in rates of chronic disease. Strategies and programs to prevent overweight among children are urgently needed. PMID- 12891869 TI - Overweight and obesity in adults and risk of renal cell carcinoma in Canada. AB - OBJECTIVES: To assess the role of overweight and obesity in renal cell carcinoma (RCC) risk in Canada. METHODS: Mailed questionnaires were used to obtain data on 1,279 (691 male and 588 female) newly diagnosed, histologically confirmed RCC cases and 5,370 population controls, between 1994 and 1997, in eight Canadian provinces. Data were collected on socio-economic status, height, weight, smoking habits, alcohol use, diet, and residential and occupational histories. Weight was expressed as body mass index (BMI). Odds ratios (ORs) and 95% confidence intervals (CIs) were derived using unconditional logistic regression. RESULTS: The study found an increased risk of RCC associated with overweight and obesity among both male and female adults; the test for trend was statistically significant (p < 0.0001 for both sexes). Compared with normal BMI, the adjusted ORs for obese class III (BMI > or = 40.00) were 3.7 (95% CI = 1.5-9.4) and 3.8 (95% CI = 2.3-6.4) among males and females, respectively. CONCLUSIONS: These findings indicate that overweight and obesity play an important role in the etiology of renal cell among both males and females. PMID- 12891870 TI - Characteristics of an Italian diabetic population and long-term changes in glycaemic control at varying body weight categories. AB - OBJECTIVES: To describe the characteristics of a diabetic population in a primary care setting, and to evaluate changes in glycaemic control at varying body weight categories. METHODS: We collected 4,610 valid baseline and three years follow-up paired data sheets sent from 77% of the general practitioners adhering to our shared-care program (257 GPs out of 521) for people with type 2 diabetes. Three conventional body weight categories were defined: "normal" (BMI, < or = 25 kg/m2), "overweight" (BMI 25.0-29.9), "obesity" (BMI > or = 30). According to weight status we calculated descriptive statistics and follow-up paired changes. RESULTS: A fifth of the diabetic patients had a "normal" BMI, about 45% was "overweight" and 34% was "obese". At follow-up, the glycaemic control significantly improved both in "normal" and "overweight" people, while it significantly worsened in "obese" ones. CONCLUSIONS: Our data confirm the crucial role played by body weight and its changes in diabetic patients. PMID- 12891871 TI - Body mass index, waist hip ratio, and waist circumference: which measure to classify obesity? AB - OBJECTIVES: To determine the proportion of a representative population sample of adults in South Australia who have a body mass index (a measure of overall obesity) classified as normal or underweight, but who also have a waist circumference or waist hip ration (measures of central obesity) that indicates obesity. METHODS: A representative population sample of adults aged 18 years and over living in the north west region of Adelaide (n = 2523) were recruited to the study. Clinical measures of height, weight, waist and hip circumference were obtained and used to determine body mass index, waist hip ratio and waist circumference. RESULTS: Among women with a normal body mass index, 19.0% had a high waist circumference (> or = 80 cm) and 8.5% had a high waist hip ratio (> 0.85). Among males with a normal body mass index, 3.4% had a high waist circumference (> or = 95 cm) and 0.1% had a high waist hip ratio (> 1.0). CONCLUSIONS: Body mass index, waist hip ratio and waist circumference all have a role in the identification of those who are obese or overweight. PMID- 12891872 TI - Appropriateness of methadone maintenance treatment for opiate addiction: evaluation by an expert panel. AB - With some 30,000 dependent persons, opiate addiction constitutes a major public health problem in Switzerland. The Swiss Federal Office of Public Health (FOPH) has long played a leading role in the prevention and treatment of opiate addiction and in research on effective means of containing the epidemic of opiate addiction and its consequences. Major milestones on that path have been the successive "Methadone reports" published by that Office and providing guidance on the care of opiate addiction with substitution treatment. In view of updating the recommendations for the appropriateness of substitution treatment for opiate addiction, in particular for the prescription of methadone, the FOPH commissioned a multi-component project involving the following elements. A survey of current attitudes and practices in Switzerland related to opiate substitution treatment Review of Swiss literature on methadone substitution treatment Review of international literature on methadone substitution treatment National Methadone Substitution Conference Multidisciplinary expert panel to evaluate the appropriateness of substitution treatment. The present report documents the process and summarises the results of the latter element above. The RAND appropriateness method (RAM) was used to distil from literature-based evidence and systematically formulated expert opinion, areas where consensus exist on the appropriateness (or inappropriateness) of methadone maintenance treatment (MMT) and areas where disagreement or uncertainty persist and which should be further pursued. The major areas which were addressed by this report are Initial assessment of candidates for MMT Appropriate settings for initiation of MMT (general and special cases) Appropriateness of methadone supportive therapy Co treatments and accompanying measures Dosage schedules and pharmacokinetic testing Withdrawal from MMT Miscellaneous questions Appropriateness of other (non methadone) substitution treatment Summary statements for each of the above categories are derived from the panel meeting and presented in the report. In the "first round", agreement was observed for 31% of the 553 theoretical scenarios evaluated. The "second round" rating, following discussion of divergent ratings, resulted in a much higher agreement among panellists, reaching 53% of the 537 scenarios. Frank disagreement was encountered for 7% of all scenarios. Overall 49% of the clinical situations (scenarios) presented were considered appropriate. The areas where at least 50% of the situations were considered appropriate were "initial assessment of candidates for MMT", the "appropriate settings for initiation of MMT", the "appropriate settings for methadone supportive treatment" and "Appropriateness of other (non-methadone) substitution treatment". The area where there was the least consensus on appropriateness concerned "appropriateness of withdrawal from MMT" (6%). The report discusses the implications and limitations of the panel results and provides recommendations for the dissemination, application, and future use of the criteria for the appropriateness of MMT. The RAND Appropriateness Method proved to be an accepted and appreciated method to assess the appropriateness of methadone maintenance treatment for opiate addicts. In the next step, the results of the expert panel process must now be combined with those of the Swiss and international literature reviews and the survey of current attitudes and practices in Switzerland, to be synthesized into formal practice guidelines. Such guidelines should be disseminated to all concerned, promoted, used and rigorously evaluated for compliance and outcome. PMID- 12891873 TI - Science-based practice for methadone treatment: another step forward. PMID- 12891874 TI - Going Swiss? PMID- 12891875 TI - Appropriateness of methadone maintenance treatment--from individual practice to evidence-based medicine. PMID- 12891876 TI - Appropriateness of methadone maintenance treatment for opiate addiction: treatment goals and effectiveness. PMID- 12891878 TI - [Medicine and the university]. PMID- 12891877 TI - Studying methadone's appropriateness: enough already! PMID- 12891879 TI - [The American health care system: origins and developments of managed care organizations]. AB - The first Health Maintenance Organizations (HMOs) appeared in the 70's in the context of care rationing. These organizations have rapidly become responsible for delivering care to the general population. Cost/effectiveness studies and other quality control programs rapidly appeared, followed by other sophisticated cost-control devices. The following article gives information on the current difficulties encountered by HMOs. Other aspects of the care given by the HMOs are also addressed. The economical and legal difficulties to which these companies were confronted have prompted them to withdraw from some market segments. Patients have become more vulnerable, despite limited attempts of reforms. PMID- 12891880 TI - [Psoriasis: pathogenesis and treatment]. AB - Psoriasis is a T-cell mediated disease. Its pathogenesis is linked to the activation of T-cells by antigen-presenting cells and secretion of cytokines able to modulate keratinocyte proliferation and differentiation. Antigens involved in psoriasis triggering are unidentified. Several new biological immunomodulatory treatments able to block T-cell activation and epidermal cells modifications are currently investigated. PMID- 12891881 TI - [Importance of allergic systems in infant consultations in the French-speaking community of Belgium]. AB - We carried out an investigation towards more than 300 families in the preventive consultations of the French-speaking Community of Belgium (O.N.E.). More specifically we chose children aged from 11 to 15 month. We used a questionnaire managed by a nurse for the parents to measure the prevalence of asthma and allergic symptoms in these children and to study their risk factors of allergies. 58% of the children have at least a family antecedent of allergy signalled and 82.1% who have a mother who presents asthma antecedents. 55.1% had an allergic symptom during there first year of life. Only 2.9% of the children had a diagnosis of asthma. Moreover the study confirms association between respiratory complaints, cigarette smoking among parents and the presence of domestic animals in the environment of the child. In the families with allergic antecedents, 80.8% respond positive on the fact of take antiallergic measures against 20.2% in the families without allergic antecedents. This work confirms the importance of allergies in preventive consultations of children and shows the necessity to follow regularly the evolution of these health problems in order to inform parents and to carry out preventive actions adapted to the knowledge and the perception of risk of each family. PMID- 12891882 TI - [Importance of echocardiography and MRI in periventricular leukomalacia]. AB - Periventricular leukomalacia (PVL) consists of focal coagulation necrosis and of more diffuse and mild white matter involvement as gliosis and lesions of immature oligodendrocytes. PVL affects essentially the premature newborn and has become the dominant form of perinatal neurological insults. Its diagnosis remains difficult to assess as neurological examination is poorly contributive. Diagnostic imaging plays an important role as it completes clinical, biological and electrophysiological data in depicting the cerebral lesions. Cerebral ultrasound remains the first imaging modality to perform in order to investigate and follow ill neonates. However ultrasound has a low sensitivity and specificity in depicting non cavitated PVL. Magnetic resonance imaging (MRI), in the acute stage, provides additional information about the extension and the depiction of hemorrhagic component of the lesions. Furthermore, diffusion-weighted MRI appears useful for the early identification of the diffuse component of PVL. MRI in the chronic stage is the only imaging method to study the progress of myelination and precise the extension of sequellar lesions. PMID- 12891883 TI - [Meglitinide analogs: new insulinotropic agents for the treatment of non-insulin dependent diabetes]. AB - In 1995, several new molecules under study as potential insulinotropic agents for the treatment of non-insulindependent diabetes mellitus were identified as analogs of meglitinide, previously known as the non-sulfonylurea moiety of glibenclamide. Three of these molecules, namely repaglinide, nateglinide and mitiglinide are or will be soon available for administration to diabetic patients. The present report aims at reviewing both preclinical studies and clinical investigations concerning the latter three meglitinide analogs. Their insulinotropic action seems attributable, like that of hypoglycaemic sulfonylureas, to a primary effect on the ATP-sensitive K+ channels of pancreatic insulin-producing cells. These meglitinide analogs differ from one another, however, by their potency as insulinotropic agents and by the time course of their biological effects, especially in terms of the reversibility of such effects. PMID- 12891884 TI - [Erectile dysfunction and phosphodiesterase type 5 inhibitors]. AB - Erectile dysfunction affects 150 millions of men and its prevalence increases with age. The improvement of life expectancy will increase the worldwide prevalence to 300 million in 2025. Oral treatments are nowadays the first line therapy for the vast majority of people as they have a good reliability and tolerance and restore more spontaneity. The authors relate the widespread interest in phosphodiesterase type 5 inhibitors with the advent of sildenafil for the treatment of erectile dysfunction and present characteristics of 2 new phosphodiesterase type 5 inhibitors in Belgium, tadalafil and vardenafil. PMID- 12891885 TI - [Sudden fainting in a young woman]. AB - A 37-year old woman (without personal medical records) is admitted in hospital for a sudden syncope (duration 15 minutes) with postcritical condition. Her family records show the sudden death of the mother (at 39). Complementary exams do not indicate anything abnormal except a hypercholesterolemia and the electrocardiogram reveals a lengthening of the QT interval. The cardiac check-up of the family members also reveals a lengthening of the QT interval of the two patient's girls. The audiometry is normal and genetic analysis shows mutations of genes coding for a cardiac ionic canal. The diagnosis of "Romano-Ward" syndrome (syndrome of long congenital QT with dominant autosomal transmission, but without deafness) is thus retained. This syndrome as well as the other form of long congenital QT (syndrome of Jervell and Lange Nielsen with recessive autosomal transmission and deafness) are caused by mutations of genes coding for cardiac ionic canals, leading to a prolonged repolarization. The basic treatment is based on betablockers medication. A pacemaker is necessary in case of severe bradycardia, whether spontaneous or resulting from betablockers medication. An implantable cardioverter defibrillator can be a resort in refractory cases. PMID- 12891886 TI - [Frederic Bremer (1892-1982): a pioneer in sleep research]. PMID- 12891887 TI - [Dental feats 1. An implant-supported orbital prosthesis after ablative tumor surgery]. AB - Extraoral implants are used with increasing frequency, in order to provide retention for facial prostheses. Facial prostheses can replace lost extraoral hard and soft tissues using acrylic or silicone materials. The surgeon, maxillofacial prosthodontist and dental technician are facing particular treatment considerations and practical problems. Some of these are demonstrated and discussed in this article by means of a case-report. A patient received an orbital prosthesis which obtains its retention from implants, placed in the lateral orbital rim. PMID- 12891888 TI - [The effectiveness of a school-based fluoride rinsing program for dental risk groups]. AB - In the Netherlands there are still groups of children whose dental health is worse compared with the majority of children. Mostly these children are from lower socio-economic strata in the larger Dutch cities. The Municipal Health Departments in Rotterdam and Amsterdam have developed a prevention programme for those risk groups. This article concentrates on the effects of fluoride mouthrinsing in children in Rotterdam who can be considered at risk as far as dental health is considered. For a period of 5 years these children participated in a fluoride mouthrinsing program at school. At three moments in time dental examinations took place: when the children were 5-6 years old, when they were 9 10 years old and finally when they were 11-12 years old. Compared to a controlgroup it appears that after 5 years the increase of dental caries in the experimental group is significantly lower than in the controlgroup. Fluoride mouthrinsing at primary schools therefore seems to be an effective form of dental prevention for groups at risk as far as dental health is considered. PMID- 12891889 TI - [Musculoskeletal disorders of the masticatory system. Consensus concerning diagnosis and therapy in gnathology]. AB - Diagnosis and management of musculoskelatal disorders of the mandibular locomotor system have changed throughout the past decades. The Dutch Society of Temporomandibular Disorders and Prosthetic Dentistry initiated a consensus on this subject. The Utrecht Medical Center and the dental schools of the universities in Amsterdam, Groningen and Nijmegen produced a consensus statement, which eventually may lead to guidelines for the Dutch dental profession. The bi axial somatic and phychosocial diagnostic approach is endorsed. Assessment, diagnostic subgroups, and management strategies, including occlusal appliances, physiotherapy, psychological treatment, and surgery are described. Counseling and reversible treatments are first choice options, although minimal invasive procedures may be considered in the early phase approach. PMID- 12891890 TI - [Dental professionals in the future: flexible team players. The vision of dental professionals on future professional structure and occupation]. AB - The project 'From care-demand for care to a social dental occupational and educational structure' was carried out to reach an agreement about the organization of the future social occupational and educational structure and the future performance of one's profession. A descriptive analysis of data, obtained by literature search and consensus-meetings was used. All relevant professional associations participated in this project. The project resulted in profiles for the future dental professionals and for the future dental team. In this project the dental professionals come to an agreement about the future professional performance. However, the discussion has not yet been finished. Further collaboration between the professional associations is recommended. PMID- 12891891 TI - [Oral health care, priorities and strategies]. PMID- 12891892 TI - [Enduring oral dryness after acne treatment]. AB - Medication influences the salivary flow rate frequently. In this paper a 26-year old patient is described, who used a systemic retinoid (a vitamin A derivate) when he was 18 years old. Since then, irreversible xerostomia was present. The oral complaints have been monitored during three years. Saliva was collected to assess the salivary flow rate and pH. The visco-elasticity of unstimulated whole saliva was high. This indicates a relatively low contribution of the gl. parotidea and a high mucin concentration in the collected saliva. Furthermore, parafilm only slightly stimulated the salivary flow rate. On the other hand, application of a 4% citric acid solution raised the flow rate to normal levels, without any delay. The medical history revealed no factors which could explain the the severe oral dryness and low salivary flow rate in rest. It was concluded that the low salivary flow rates and xerostomia might be related to the previous use of isotretinoin (Roaccutane). It is suggested to register and monitor the use of medication in patients with sudden oral health changes. PMID- 12891893 TI - [Glaucoma]. AB - Glaucoma is a group of afflictions that have loss of eyesight in common. This impairment of vision is caused by damage to the optic nerve as a result of high intraocular pressure. The patient's optical fields narrow and at last only a tiny rest is left. The mechanisms of the disease are well known but it is in most cases unexplained why a person develops glaucoma. This review discusses the several forms of glaucoma, the diagnosis and therapy, and the epidemiology and prevention. PMID- 12891894 TI - [Enantioseparation of quinazolone derivatives using an AGP-based liquid chromatography stationary phase]. AB - An AGP-based chiral stationary phase has been applied successfully for enantioseparation of 4(3H)-quinazolone derivatives. Chiral-AGP is proved to be an excellent selector, as optimized chromatographic conditions allow with one exception baseline resolution for the enantiomers of the potential cholecystokinin antagonist compounds (alpha = 1.19-1.85). Retention and enantioselectivity could be modified to a large extent by varying the eluent pH and adding organic solvents with different types i.e. acetonitrile and 2-propanol to the buffered mobile phase. It was established that by increasing the eluent pH from 6.0 to 7.0 the retention factors of of the model compounds bearing no protonable groups are increased in the presence of 7 v/v % (1.33 M) acetonitrile. However further increasing the acetonitrile content up to 10 v/v % or addition of 2-propanol in equimolar concentration (1.33 M) no similar changes could be detected with the same modification in the eluent pH. These observations are explained by changes in the sorption properties of the selector determined simultaneously by the type and concentration of organic modifier and also the eluent pH. The experimental data give further insight into the chromatographic mechanism on a Chiral-AGP column. PMID- 12891895 TI - [Fast liquid chromatographic method to determine compounds binding to human serum albumin]. AB - A fast gradient HPLC method (cycle time 15 min) has been developed to determine Human Serum Albumin (HSA) binding of discovery compounds using chemically bonded protein stationary phases. The 2- propanol concentration is increased from 0 to 25% in 3 minutes, thus allowing strongly bond compounds to elute within the 15 minutes. The HSA binding values were derived from the logarithmic values of the gradient retention times that were converted to logarithm of the equilibrium constants (logK HSA) using a calibration set of molecules. The logK HSA values for the calibration set of molecules have been derived from literature values of the % plasma protein binding. The method is fully automated and it has been used for lead optimization in more than 20 projects. The obtained HSA binding data on more than 4000 compounds were suitable to set up global and project specific quantitative structure binding relationships that helped compound design in the early drug discovery settings. The method has been validated using literature plasma protein binding data of 70 known drug molecules. It was shown that compound lipophilicity dominates the HSA binding, however compounds binding to a specific binding site show stronger HSA binding that can be expected from their lipophilicity values. The solvation equation approach has been used to characterize the non-specific lipophilic binding ability of HSA. Based on our approach we can identify compounds with specific and non-specific binding. PMID- 12891896 TI - [Capillary electrochromatography for chiral separation and purity testing of pharmaceutical drug substances]. AB - Chiral separation of D,L-Norgestrel was successfully performed by cyclodextrin modified capillary electrochromatographic (CEC) method. Optimized CEC conditions for purity testing were achieved: Hypersil C-18 packed capillary and 10 mM 2-[N morpholino]-ethanesulfonic acid buffer (MES) and 40 v/v % methanol as background electrolyte containing 10 mM gamma-cyclodextrin at pH = 5.5. The resolution was found to be strongly dependent on the concentration of chiral selector gamma cyclodextrin, applied voltage and temperature of the capillary. Our results represent a simple, fast and reproducible method for the separation of D,L Norgestrel and purity testing of D-Norgestrel. It can be concluded that cyclodextrin modified HPLC, HPCE and CEC offer inexpensive, attractive and reliable chiral separation methods. PMID- 12891897 TI - [The multidrug resistance modulators heterocondensed quinazolones]. AB - Exploration for new MDR-modulators utilizing pyrazino[2,1-b]quinazolones as scaffolds disclosed after systematic synthetic investigation highly hydrophobic N substituted derivatives as a readily accessible active tricyclic compounds. A versatile synthesis of 2-substituted-1,2,3,4-tetrahydro-6H-pyrazino[2,1 b]quinazoline-3,6-diones is presented starting from 2,3-substituted quinazolines. The new compounds have been characterized by elemental analyses, 1H nmr and in some cases by 13C ruler, and X-ray investigations. PMID- 12891899 TI - [The effect of physical aging on the structural behavior of amorphous poly(vinylpyrrolidone) polymers]. AB - The purpose of the present study was to examine the physical ageing of amorphous polymers commonly applied in the pharmaceutical technology. Powder samples of polyvinylpyrrolidone were stored under different storage conditions (relative humidity, temperature). Positron annihilation lifetime spectroscopy (PALS) was applied for the analysis of the free volume changes of polymer samples as a function of the storage conditions. By the means of calculations on simplified model molecules it was determined that polymer chains and water molecules are able to form a hydrogen bound "network" under certain humidity conditions. Along with the ageing process the chemical changes of the examined polymers are interrelated in a dynamic and complex manner which can be tracked by the combination of SEM, ab initio calculations and positron lifetime measurements. PMID- 12891898 TI - [Light- and temperature-induced degradation of fumagillin]. AB - The antibiotic fumagillin with amebicidal and fungicidal effects isolated from Aspergillus fumigatus is the only presently known agent for the treatment of life threatening serious microsporidiosis occurring in patients with AIDS. Fumagillin and its degradation products were measured by HPLC at given time intervals after storage under defined conditions (temperature, relative humidity). Significant degradation took place even in samples stored in freezer; therefore fumagillin drug substance should be stored below -60 degrees C. Light also induced a degradation process in fumagillin, thus it is proposed to be stored and transported in brown glass. PMID- 12891900 TI - [Synthesis of dicarboxylic acid monoamides]. AB - The pharmaceutical and biological importance of the amide moiety is briefly surveyed. Relationships between the electron density and chemical reactivity of the amide site are shortly described. Synthetic methods for the selective formation of monoamino-dicarboxylic acid alpha and beta monoamides are summarised. The three major selective synthesis routes for the preparation of monoamides are introduced. The first class of synthetic methods consists of non selective formation of a- and b-esters, followed by their separation on the basis of different solubilities. Amidation of these esters results in the alpha- or beta-amides respectively. The second class of the reactions utilises the orientating capacity of the amino (ammonium) site, producing first various cyclic anhydrides, lactames, lactones, which are then decomposed in hydrolytic, aminolytic, etc. reactions, resulting selectively in the alpha- or beta-amides or esters. Reactants in the third class of the reactions are dicarboxylic acids with carbon-carbon pi-bonds, and ammonia (or alkyl-amine) which form the appropriate compound in addition reactions. Reactivities and selectivities are interpreted in terms of inductive effects, acidity differences and electronic effects of the various protecting groups. Some important analytical properties of monoamino dicarboxylic acids and their monoamides are compiled. PMID- 12891901 TI - [Development of percutaneous coronary intervention in Denmark]. PMID- 12891902 TI - [Percutaneous coronary intervention in Denmark from 1989 to 1998. Results from the Danish PTCA registry]. AB - INTRODUCTION: During the ten-year period 1989 to 1998, all percutaneous coronary interventions (PCI) in Denmark were recorded in the Danish PTCA Registry. The purpose was quality control based on treatment activity, severity of illness, technique employed, success rate, and complications. MATERIAL AND METHODS: For each PCI procedure a number of 144 parameters were recorded in a national database. The present analysis of these data was carried out using parametric statistics and logistic regression analysis. RESULTS: From 1989 to 1998 the annual number of PCI procedures increased from 240 to 3840. The database contains 13,868 procedures of which 10,804 are first time interventions. Men accounted for three fourths of the patients. The mean age was 60.7 years for women and 57.9 years for men. During the ten-year period a rise of two years on average was seen for both sexes with a doubling of patients being older than 70 years. More patients with acute coronary syndrome and multivessel disease had PCI. The use of coronary stents increased from 2.0% in 1992 to 90.7% in 1998 and the primary success rate rose from 81% to 90%. Serious complications decreased from 5.4% to 3.0%, mainly due to a significant fall in procedurally related acute bypass surgery. Mortality remained unchanged around 0.6%. Age was found to be the only independent predictor for major procedurally related complications. DISCUSSION: From 1989 to 1998 a sixteenfold increase in the number of PCI was observed. In parallel to this was seen an increase in the use of coronary stents and in the procedural success rate, while the frequency of serious complications decreased. This progress is similar to that of several other European countries. PMID- 12891903 TI - [Surgical services in Denmark--time for reorganization?]. AB - Denmark is a small country (about 5 million inhabitants) and has a unique registration of hospital admissions, surgical procedures and specific person numbers in national registers. However, there is currently no system to monitor the overall quality of surgical care. Also, surgical services are provided in a large number of hospitals. In order to provide a rational basis of professional and administrative strategies to monitor and improve the quality of surgical care, establishment of a nation-wide monitoring system and quality improvement programme is required as well as regionalisation of major procedures. PMID- 12891904 TI - [Vocational rehabilitation, a descriptive follow-up study]. AB - INTRODUCTION: Only few studies of the long-term occupational situation of clients after stay in a vocational rehabilitation clinic have been published. The aim of this study was to describe the clients in a specific vocational rehabilitation clinic, to investigate the employment situation 6-10 months after their stay, and to find out whether a doctor's estimate can point out the clients who are able to return to employment, and who are not. MATERIAL AND METHODS: All 254 clients that commenced a stay in the rehabilitation clinic from May 1996 to April 1998 were included. A doctor estimated whether he thought the client could return to employment or not. After 6-10 months the clients received a mailed questionnaire. A total of 210 persons (82.7%) answered the questionnaire. RESULTS: Eighty per cent of the clients were women. About two thirds of the clients were unskilled. Musculoskeletal disease was the reason of vocational disability in 63% of the clients. At the follow-up 50% were awarded disability pension, 20% were either employed or still in a rehabilitation process, whereas 30% still did receive passive support. If the doctor estimated that the person could not return to employment, the positive predictive value was 94.1-98-8%. If the doctor estimated that the person could return to employment the positive predictive value was 36.1 63.9%. DISCUSSION: Many of the clients that were later awarded disability pension could at an early time be pointed out by a doctor. These clients might not need to go through a stay in a vocational rehabilitation clinic. The methods used in vocational rehabilitation clinics should be evaluated and developed in order to make more clients return to employment. PMID- 12891905 TI - [Lactation--a risk factor for elevated plasma homocysteine?]. AB - INTRODUCTION: Women have an increased need of folate in the breastfeeding period and, as a consequence, may be in risk of having a negative folate balance. Elevated total homocysteine in plasma (P-tHcy) is a sensitive marker for deficiency of folate, B6, and B12 vitamins and a risk factor for developing cardiovascular disease. We examined whether there is a difference in P-tHcy among breastfeeding mothers depending on folic acid supplement and compared these results to women from the background population. MATERIAL AND METHODS: A total of 20 breastfeeding mothers had their plasma homocysteine content and their erythrocyte folate content measured through a standard blood sampling procedure in their homes. Half of the mothers took a folic acid supplement. The background population was age-matched, non-pregnant, and non-lactating women from a previous Danish study. RESULTS: The group of breastfeeding mothers who did not take folic acid supplements had a higher prevalence of elevated P-tHcy (70%) both than the group of breastfeeding mothers who took folic acid supplements (30%) and than the background population (33%). The difference between P-tHcy in the group of breastfeeding mothers who did not take folic acid supplements and the group of breastfeeding mothers who took folic acid supplements was non-significant (p = 0.12). However, we found a statistically significant difference between the group of breastfeeding mothers who did not take folic acid supplements and the background population (p = 0.02). DISCUSSION: Our study suggests that there is a significant fraction of Danish breastfeeding mothers in negative folate balance resulting in elevated P-tHcy compared to the background population. PMID- 12891907 TI - [Continuing education meeting]. PMID- 12891906 TI - [Thromboembolic complication following temporary discontinuance of anticoagulant therapy in a patient with mechanical heart valve]. AB - Patients with mechanical heart valves are treated with oral anticoagulant therapy in order to minimize the risk of thromboembolism. During invasive procedures it is recommended to temporarily discontinue the oral anticoagulant therapy and substitute with injections of low-molecular-weight heparin. We describe a case where a patient discontinued the oral anticoagulant therapy and had no substitution with low-molecular-weight heparin. The patient had an embolism to the coronary arteries causing progressive heart failure, and the patient was subsequently heart transplanted. PMID- 12891908 TI - [Stem cells and ethics: the new law proposal is not extensive enough]. PMID- 12891909 TI - [The prostate cancer in the community of Madrid in 2000 I.- Incidence]. AB - OBJECTIVE: To know the incidence in the year 2000 of prostate cancer in the Autonomous Community of Madrid and its breakdown by Health Areas. MATERIAL AND METHOD: Study of histologically confirmed prostate cancer case reports and retrospective data acquisition for 2000 in the Autonomous Community of Madrid, both from Public and Private Health Care hospitals. RESULTS: Gross incidence of prostate cancer in the Autonomous Community of Madrid was 100.4 cases per 100,000 males. The incidence adjusted for the Spanish, European and Worldwide population was 120.1, 103.5 and 68.6 cases per 100,000 males, respectively. Mean age at diagnosis was 70 +/- 7.8 (40-94) years, median of 70 years. The age group with higher incidence was 70 to 79 years. CONCLUSIONS: The incidence of prostate cancer in the Autonomous Community of Madrid is lower than that in the US but higher than in most countries or regions in the EU. The different way of using PSA testing in the Health Areas of the Autonomous Community may explain the differences seen in terms of incidence by Area. PMID- 12891910 TI - [The prostate cancer in the community of Madrid in 2000. II Presentation and diagnosis]. AB - OBJECTIVE: To know the presentation form, diagnostic method and clinical stage at the time of diagnosis in subjects with prostate cancer (PC) in the Autonomous Community of Madrid in 2000. MATERIAL AND METHOD: Data from 1745 patients with histologically confirmed prostate cancer obtained from 15 Hospitals participating in the study was analysed. The variables studied were: associated disease, reason for visiting the hospital, digital rectal examination (DRE), PSA, diagnostic method, graded Gleason score, tests performed in the tumoral extension study and tumour staging. The qualitative variables are given in percentages of the overall number and the quantitative variables are expressed as the median, standard deviation, maximum and minimum values and 25%, 50% (median) and 75% percentiles. RESULTS: 67% cases had an associated disease. In most (75%) patients the reason for visiting the hospital was prostatic syndrome. DRE revealed that 42.7% has no tumour. At the time of diagnosis half the patients had PSA levels lower than or equal to 11 ng/ml. Transrectal ultrasound-guided biopsy was used for diagnosis in 93% subjects. The most commonly reported Gleason scores were 6 (31.3%) and 7 (28.7%). In 75% subjects the disease was considered to be clinically limited to the prostate, in 12.5% locally advanced and in 12.5% metastatic. CONCLUSIONS: Most patients came to the hospital because of symptoms not related to PC. Transrectal ultrasound-guided biopsy is confirmed as the choice technique for PC diagnosis. When a comparison is made to historical series in our Autonomous Community a pattern of earlier diagnosis can be seen. PMID- 12891911 TI - [Clinical and biological progression of incidental prostatic cancer ( Stage T 1a)]. AB - OBJECTIVES: To analyze the biological and clinical progression of the prostate cancer stage T1a. MATERIAL AND METHODS: Retrospective study of 44 patients diagnosed of T1a prostate carcinoma between 1985 and 2001. We value biological and clinical progression, time up to the progression, mortality for tumour reason and survival, with the following stratification: patients without initial treatment and patients treated by means of external radiotherapy or radical prostatectomia. RESULTS: Of all 44 patients biological progression was observed in 5 (11.36%) and clinical progression in 4 (9.09%). The mortality to 5 years for tumour reason was of 2 (4.54%). Of all 38 patients without initial treatment biological progression was observed in 5 (13.15%), in an average time of 25.8 months and clinical progression in 4 (10.52%), in an average time of 34.5 months. The mortality to 5 years was of 2 (5.26%). In all 6 patients to whom radical treatment carried out them progression was not observed and they all live. There are no statistically significant differences between both groups of patients (p = NS). CONCLUSIONS: The biological and clinical progression of the T1a prostate cancer is low, 11.36% and 9.09%, respectively. The mortality to 5 years is of 4.54%. Differences of survival do not exist, statistically significant, between treated and not treated. PMID- 12891912 TI - [Cystectomy in superficial bladder tumours. Analysis of criterion include and prediction factors]. AB - INTRODUCTION: Though usually the treatment of a superficial bladder tumour consists in transurethral resection, exceptionally because of several clinical conditions, in this kind of patients a radical cystectomy may be suitable. PATIENTS AND METHOD: From june 1986 through december 2001, 127 radical cystectomies were performed. A series of 25 patients with a radical cystectomy performed in superficial bladder tumours were analysed. Indications, anatomo pathological correlation and clinical evolution with survival curves are analysed. RESULTS: The mean time of follow-up was 47 months. Extensive GIIIpT1 tumours (40%) were the chief indication. The remaining of the series consisted in carcinoma uncontrollable by endoscopy and refractory to chemotherapy. Anatomo pathological correlation coincided in 48% of patients, existing supra and understaging in 25% and 28% respectively. A period of 54 months was without relapse. Seven patients died during the evolution (28%), and 18 patients are alive without disease. 128,48 months were the mean actuarial survival. CONCLUSIONS: GIIIpT1 tumours were the chief indication of our series. The significant percentage of understaging, poor morbidity and good survival curves are emphasized. Because of these results we consider that radical cystectomies are a viable choice for selected patients with superficial vesical tumour. PMID- 12891913 TI - [Predictive model for prostate cancer in patients with biopsy indication]. AB - OBJECTIVE: Attemp to determine the probability of developing prostate carcinoma taking into acc age, digital rectal examination and PSA once a transrectal biopsy has been indicated, so that both doctors and patients have mor information to face such pathology. MATERIAL AND METHODS: Retrospective study of 633 biopsies, taken into acc the patient's age, digital rectal examination, PSA level and histology. The data were included in a database created with Access and were put a logistic regression by mens the software program SPSS. RESULTS: Once the biopsy is indicated, digital rectal examination is the parameter offesing a higher discriminatory valuer with an odd ratio of 5.9 (CI 95%, 3.9-8.9). The mathematical model obtained shows a sensitivity level of 57% and a level of specificity of 84%. Pre-test probability is 36%, the probability post-test increasing up to 70%, and a negative predictive value of 77% and a positive predictive value of 67%. CONCLUSIONS: The mathematical model obtained individually determines the probability of suffering from prostatic carcinoma. Moreover, using this model the probabilities obtained re more precise than those derived from the fact of fulfilling the criteria for a prostatic biopsy. Once a biopsy is indicated, the rectal examination becomes the parameter with a higher predictive value of PC, irrespective of PSA and age. The PPV of the model is higher than of the PSA or the digital recta examination used separately. PMID- 12891914 TI - [Effects of water diuresis on the urinary ph of patients under relapsing lithiasis]. AB - INTRODUCTION: It has been observed that in healthy individuals the increase in urinary flow induced by water overload may be enough to alter the urinary Ph to obtain values considered safe in the prophylaxis of lithogenesis. The aim of the present paper is to determine the effects of water diuresis on the urinary Ph of patients suffering from relapsing lithiasis. MATERIAL AND METHOD: 26 patients were included in the present study. The urinary Ph of all patients was measured along two consecutive days, at 8.30 and again at 12.30, under normal conditions (the first day) and once they had received a water load equivalent to 1.5% of their weight (the second day). RESULTS: The altered mean value of the Ph induced on the first day by the circadian rhythm showed a mean of 0.13 units of Ph, which lacks statistical significance (p > 0.05). However, on the second day we obtained a value of 0.42, which is statistically significant (p < 0.01). The normal tendency after the water overload was towards a positive increase of the Ph in those patients whose urine showed a preload Ph value < 6.1, whereas those individuals with a preload Ph value > 6.48 suffered negative increases. CONCLUSIONS: The increase of diuresis induced by water load proved enough to provoke an increase of urinary Ph when its baseline value is < 6.1, or either a decrease when its baseline value is > 6.48. In both cases, Ph values ranged between 6-6.5 which are considered safe values in the prophylaxis of lithogenesis. The more separated the baseline urinary Ph is from the 6.1-6.48 range, the greater the effect of the water load. PMID- 12891915 TI - [Laparoscopic radical prostatectomy. Preliminary experience]. AB - OBJECTIVE: Laparoscopic technique has been developed with the aim to decrease the morbidity of the open radical prostatectomy. MATERIAL AND METHODS: From january 2002 to may 2002, 8 patients were treated for prostate cancer with laparoscopic radical prostatectomy. Unilateral linfadenectomy has been carried out in only one patient. We usually employ the transperitoneal technique published by the Montsouris Institute, with some modifications. RESULTS: The main surgical time was 356 minutes (540-240). Transfusion wasn't needed in any case. Intraoperative complications were: 2 bladder injuries, 1 bleeding of the epigastric artery. Postoperative complications were: 1 ileus, 2 compressive neurapraxia, 4 anastomotic fistutas. All complications were treated conservatively. No patient were converted to open surgery. Surgical limits were negatifs in all cases, and the PSA rate was less than 0.1 ng/ml in the first month follow up. CONCLUSION: Laparoscopic radical prostatectomy is a difficult technique, but we think that, the learning curve is getting lower and lower than in the first series. Oncologic results with this technique is similar to the open one. However, laparoscopic approach shows us some benefits such as less bleeding, less time catheterisation, less hospital stay, better continence, better sexual function, better stetic result, less postoperative pain, and finally an earlier back to work. PMID- 12891916 TI - [Benign retroperitoneal schwannoma in patient with antecedents of Willm's tumor in the infancy]. AB - We report a case of benign retroperitoneal schwannoma diagnosed casual in a 23 years old woman treated of a Wilm's tumour twenty years ago. A review of the on this unusual tumour is included. PMID- 12891917 TI - [Dedifferentiated liposarcoma of spermatic cord: degeneration of lipoma previously resected]. AB - Spermatic cord sarcomas are rare tumours. Dedifferentiated liposarcoma accounts for only 10% of all spermatic cord sarcomas. These are usually large-sized tumours histologically characterised for being well-differentiated liposarcomas with some high grade sarcoma areas. Volume, location, mass homogeneity as well as presence of pelvic and retroperitoneal adenopathies are reported by CT and ultrasound techniques. These are useful for post-treatment follow-up. This paper presents one spermatic cord, dedifferentiated liposarcoma from which lipomas from the same spermatic cord had been previously removed in three occasions. We believe this is a degeneration of the earlier resected lipoma. Management is by extended inguinal radical orchiectomy. Value of adjuvant radio- and chemotherapy is uncertain. Post-surgery local relapses are common, and haematogenous and pelvic nodes metastasis likely. Survival at 5 and 10 years is 75% and 63% respectively. PMID- 12891918 TI - [Renal metastasis of hepatocellular carcinoma]. AB - OBJECTIVE: To report the case of a solitary renal metastasis secondary to hepatocellular carcinoma. METHODS: We report the case of a 51 year old patient who on abdominal ultrasonography was revealed a left renal tumour and a hepatic mass incidentally. A TAC showed the left renal tumor measuring 17 cm in size, possible involvement of left renal vein and a tumour mass in the right lobe of the liver. A TAC guided fine needle punction aspiration biopsy demonstrated a malignant hepatic lesion compatible with hepatocarcinoma, and malignant renal cells compatible with renal or adrenal carcinoma. Left radical nephrectomy and right hepatectomy was performed. RESULTS: Histopathologic study confirmed the diagnosis of moderately differentiated trabecular hepatocarcinoma with lymph node and left renal metastasis. PMID- 12891919 TI - [Hypernephroma in native kidney of patient with renal transplant]. AB - The uremic patients, specially in periodic dialysis, may present bilateral kidneys cytes with high frequency. This pathology, known as acquired cyst renal disease, is found associated in its evolution with the emergence renal malignant disease. A patient who undergoing chronic dialysis and renal transplant subsequently developed an asymptomatic renal neoplasm. The epidemiological, clinical, diagnostic aspects and therapeutic of the acquired cysts renal and your association with renal cancer are analyzed. PMID- 12891920 TI - [Bladder endometriosis. Diagnostic and treatment]. AB - Endometriosis is a common gynecologic disease in which endometrial tissue is deposited outside the normal confines of the uterine cavity. Rarely endometriosis involves urinary tract. The bladder is the most frequent organ affected into this tract. A delay in detection can result in months or years of morbidity. PMID- 12891921 TI - Prevalence of microscopic colitis in patients with symptoms suggesting irritable bowel syndrome. AB - GOALS/BACKGROUND: Irritable bowel syndrome is a common disorder affecting 20% of the general population. It shows certain characteristics with organic bowel diseases. Definition of lymphocytic and collagenous colitis has created a new approach towards chronic idiopathic diarrheas. We searched for the frequency of lymphocytic and collagenous colitis in patients with irritable bowel syndrome. STUDY: The study group consisted of 30 irritable bowel patients and 20 controls. Multiple biopsies from cecum; ascendant, transverse and descendent colon; sigmoid and rectum were taken sequentially in all patients. RESULTS: We diagnosed 7 out of 30 irritable bowel patients as having lymphocytic colitis (23.3%) but none as having collagenous colitis. In the control group 1 out of 20 patients had lymphocytic colitis (5%) and none had collagenous colitis. Irritable bowel patients had higher rate of lymphocytic colitis association (p < 0.05). CONCLUSIONS: Functional disorders of the bowel should be searched for possible lymphocytic colitis, especially in cases refractory to classical therapies. PMID- 12891922 TI - An ultrastructural investigation in stomach epithelial cells of mice during pregnancy and early lactation. AB - In this study, stomach epithelial cells from adult nonpregnant, pregnant lactating mice were examined by histochemical, immunohistochemical and ultrastructural methods. Tissue samples taken from different groups were stained with haematoxylin-eosin and PAS-alcian blue and semithin sections were stained with toluidin blue for light microscopic examination. For ultrastructural examination ultrathin sections were stained with uranyl acetate and lead citrate. Additionally, by applying immunoperoxidase method, the distribution of EGF receptors of all groups was studied. We concluded that in the pregnant mice, stomach surface had lower mucus than the nonpregnant group. The acid content was balanced in active and inactive parietal cells. Examination results of lactating mice were similar to the results obtained from nonpregnant and pregnant mice. PMID- 12891923 TI - Features of chronic inflammation at the gastric cardia and the relationship with Helicobacter pylori infection and oesophagitis. AB - BACKGROUND: The etiopathogenesis of chronic inflammation at the gastric cardia is still debated. It is suggested that carditis may be a finding of gastro oesophageal reflux disease (GORD) or it may occur as a result of the gastritis caused by Helicobacter pylori (H. pylori) infection. AIM: To examine morphological features of carditis, as well as the associations of carditis with Helicobacter pylori gastritis and oesophagitis as a marker of gastro-oesophageal reflux disease. PATIENTS AND METHODS: Endoscopic biopsy specimens obtained systematically from oesophagus, cardia, corpus and antrum of 135 dyspeptic patients were retrospectively evaluated. In biopsies, we have searched for any correlations between clinical, endoscopic, and histological features. RESULTS: Carditis was detected in 123 (91.1%) of the cases. The mean age of the carditis group was 47.9 years and the male-to-female ratio was 1.08:1. The relation of carditis with age and sex was not significant (p = 0.19 and p = 0.24, respectively). All cases of the carditis group had concomitant chronic gastritis. In these cases, chronic inflammation, degree of neutrophil-mediated activity and H. pylori colonisation were significantly correlated in cardia, corpus and antrum (p < 0.001). Intestinal metaplasia was observed in 14 cases (11.3%) and, was associated with H. pylori colonisation (p < 0.001). Microscopic oesophagitis detected in 37.7% cases also showed correlation with reflux symptoms and endoscopic oesophagitis but not carditis. When all cases with carditis were evaluated for H. pylori infection and oesophagitis, which are presumed risk factors for carditis, H. pylori infection appeared to be an independent risk factor for carditis (p = 0.012), while oesophagitis did not. CONCLUSIONS: This study suggests that carditis is commonly found in patients presenting with dyspepsia and the histological features of carditis were similar to those seen in H. pylori gastritis in antrum and corpus. In addition, our data have also shown that carditis was significantly associated with H. pylori infection but not with symptoms or signs of GORD. PMID- 12891924 TI - An update on acute colonic pseudo-obstruction (Ogilvie's syndrome). PMID- 12891925 TI - Diagnosis and therapeutic problems of primary sclerosing cholangitis. AB - Primary sclerosing cholangitis (PSC) leads to a progressive destruction of the intra- and extrahepatic bile ducts. The cause is unknown but genetic and immunological mechanisms may play a role. The median survival time from diagnosis to death is about 12 years. MRCP is almost equal to ERCP for diagnosing PSC and shows the typical localised or multifocal strictures and interfering segments of ectatic bile ducts. Liver histology can be helpful in making the diagnosis but is often unspecific and there is a large sampling variability. The treatment of PSC is disappointing. The combination of ursodeoxycholic acid with endoscopic dilatation is probably the best treatment. Patients with cirrhosis and/or recurrent cholangitis should be evaluated for liver transplantation as the outcome after liver transplantation is good, especially if there is no cholangio carcinoma present and if the Child-Pugh score is not too high. There is also a need to treat the complication of PSC such as osteoporosis, cholangitis and the development of cholangiocarcinoma. PMID- 12891926 TI - Biopsy of focal liver lesions: guidelines, comparison of techniques and cost analysis. AB - When a focal liver lesion is discovered, differentiation between a benign and malignant nature and further characterization are mandatory to guide further treatment. Histology remains the golden standard. Improving imaging techniques such as contrast enhanced Doppler ultrasonography, spiral CT and new MRI procedures are promising, but not 100% accurate. When there is any doubt, biopsy should be performed. Fine Needle Aspiration Biopsy (FNAB) has a high sensitivity and specificity (90-95%) in experienced hands, but has a high insufficient sampling rate (up to 15%). In a series of 245 Fine Needle Tru-cut Biopsies (FNTCB) of focal solid liver lesions performed at our institution, sensitivity and specificity for the diagnosis of malignancy were 86% and 100% respectively, with an overall accuracy of 88%. Positive predictive value was 100%, but negative predictive value was rather low (56%). Insufficient sampling rate was low (2.5%), and a more accurate histological characterization was possible compared to FNAB. Finally, the cost-analysis of different biopsy techniques is presented for the Belgian situation according to used materials, pathology procedures and hospitalization. PMID- 12891927 TI - Pathophysiology of acute pancreatitis: a multistep disease. PMID- 12891928 TI - Early severity indexes in acute pancreatitis. PMID- 12891929 TI - Early multi-system organ failure associated with acute pancreatitis: a plea for a conservative therapeutic strategy. AB - The mortality of severe acute pancreatitis still ranges between 10 and 20%. Nowadays, infected pancreatic necrosis is the leading cause of death. Despite advances in intensive care therapy, however, early and worsening multi-system organ failure remains a source of substantial morbidity and still accounts for 20 to 50% of the deaths. In recent years, the systemic inflammatory response syndrome and the relevant cascades of inflammatory mediators have been implicated as the key factor in the emergence of remote tissue damage. Early multi-system organ failure that supervenes in the first week is typically associated with a sterile necrotizing process. There are no pathophysiological, clinical or economical data to support the practice of debridement of sterile necrosis to prevent or to control early multi-system organ failure. This issue has never been addressed in a controlled study. Besides intensive care support, non-surgical therapeutic modalities including urgent endoscopic sphincterotomy for impacted stones, antibiotic prophylaxis for the prevention of pancreatic infection and early jejunal nutrition have been specifically developed hopefully to attenuate multiple organ failure, to obviate the need of surgical drainage and to improve survival. Fine needle aspiration of necrotic areas must be incorporated in any conservative therapeutic strategy in order to identify and not to jeopardize those with infected necrosis that remains an absolute indication for drainage. A specific treatment of acute pancreatitis is still lacking, so far. However, there is ample experimental and pathophysiological evidence in favour of immunomodulatory therapy in severe acute pancreatitis. The administration of one or several antagonists of inflammatory mediators possibly combined with a protease inhibitor may at last provide the opportunity to interfere with the two major determinants of prognosis: the severity of multiple organ failure and the extent of necrotic areas that creates the culture medium for bacterial superinfection. These benefits remain to be substantiated in a controlled study, however. PMID- 12891930 TI - Endoscopic therapy of chronic pancreatitis. PMID- 12891931 TI - Primary squamous cell carcinoma of the stomach: a case report. AB - Primary squamous cell carcinoma (SCC) of the stomach is a very rare tumour. We report a case of SCC of the stomach in a 56-year old woman who had metastases to the liver and abdominal lymph nodes. PMID- 12891932 TI - Ischaemic necrosis of the rectum and sigmoid colon complicating systemic lupus erythematosus. AB - The case of a 37-year-old Caucasian female with a history of systemic lupus erythematosus, admitted to hospital due to progressively worsening abdominal pain and arthralgia. During hospitalisation, signs of acute abdomen developed. Laparotomy revealed perforation of the rectum, accompanied by necrosis of the rectosigmoid. Histologic examination revealed vasculitis involving small- and medium-sized vessels. This case report emphasizes the point that colonic and especially rectal involvement from vasculitis, though unusual, may present with profound and possibly life-threatening manifestations and stresses the difficulties in clinical and histological differential diagnosis from other causes of systemic lupus erythematosus-associated abdominal pain. PMID- 12891933 TI - [Dent disease. History and genetic causes of a tubulopathy]. PMID- 12891934 TI - [The functions of the community nephrologist and its importance in health care system]. PMID- 12891935 TI - [Clinical trials in renal transplantation: many and few. Analysis of the evidence]. PMID- 12891936 TI - [Study of the renal acidification capacity in children diagnosed of idiopathic hypercalciuria]. AB - OBJECTIVE: To study the capacity of renal acidification in a group of children diagnosed of idiopathic hypercalciuria. PATIENT AND METHODS: 36 children were studied, to those that were determined the pCO2 (UpCO2) maximum urinary with two different stimuli, acetazolamide and sodium bicarbonate (NaHCO3). At 33 of them, was performed an acidification test with frusemide stimulus. We studied a control group of 13 healthy children so much for the first one as the second tests and other 14 healthy children for the acidification test with frusemide. RESULTS: In the tests performed with NaHCO3 and acetazolamide stimulus, they were not proven differences in the values of UpCO2 neither in the urinary concentration of HCO3- (UHCO3-) than control children. Nevertheless, the UpCO2 and the concentration of UHCO3- in the patients were significantly lower with acetazolamide with regard to the NaHCO3 stimulus. In the acidification test with frusemide, significantly lower values of titratable acid and ammonium were obtained than control children. CONCLUSIONS: In children with idiopathic hypercalciuria, the capacity of secretion of H+ is normal, what is evidenced, especially, when studying the maximum UpCO2 after stimulus with NaHCO3. When diuretics are used as stimuli, exists more negative results that can be due to a certain partial resistance to the action of the same ones or to that are less potent to induce the secretion of H+. PMID- 12891937 TI - [Prognostic markers determining the start of dialysis]. AB - BACKGROUND: It is widely known that renal disease progresses towards the terminal stage regardless of the cause. The aim of this study is to identify prognostic factors in the progression that determine the start of dialysis. METHODS: From january 1998 until december 1999, 76 patients diagnosed with renal failure were monitored, 52 of whom started dialysis treatments. Clinical and analytical variables were studied. An actuarial analysis was carried out following the methodology of Kaplan-Meier to determine the likelihood of the need for dialysis and the Cox proportional risk analysis was also used. RESULTS: The patients enrolled in this study were between 20 and 78 years of age. 49 (64.5%) of the subjects were men and 27 (35.5%) were women. The most prevalent pathology was nephroangiosclerosis (26.3%), followed by diabetes (25.0%). During the monitoring program, 68.4% of the patients began dialysis treatments (table I). Systolic blood pressure, haemoglobin and the total proteins were univariately identified as prognostic variables. 89.47% and 71.58% of the patients with a systolic blood pressure reading of under 140 mmHg had not begun dialysis after 12 and 24 months respectively. 87.02% and 53.63% of the patients with a systolic blood pressure reading of over 140 mmHg had not begun dialysis after 12 and 24 months respectively (p = 0.025) (fig. 1). With a haemoglobin level higher than 10 gr/dl after 12 months 92.7% had not started dialysis, dropping to 72.33% after 24 months. With a haemoglobin level of under 10 g/dl 78.6% and 37.59% had not started dialysis after 12 and 24 months respectively (p = 0.0008) (fig. 2). Taking into account the mean values of different variables in the two year period prior to starting or not starting dialysis, it was found that haemoglobin significantly affected the risk of the need to start dialysis treatments [RR = 0.729; 95% CI = (0.554;0.959)], while systolic blood pressure was on the borderline of statistical significance (table IV). CONCLUSIONS: Haemoglobin levels significantly affect the risk of the need to start dialysis treatments, while other variables were identified as possible prognostic factors. PMID- 12891938 TI - [Influence of early or late referral to nephrologist over morbidity and mortality in hemodialysis]. AB - OBJECTIVE: We studied the influence of early vs late referral to nephrologist of patients with chronic renal failure over clinical situation at the onset of hemodialysis and outcome. SUBJECTS AND METHODS: From january 1994 to december 1998, 139 patients started hemodialysis for end-stage renal disease at the Hospital General de Albacete, all of them included in the study and clinical follow-up concluded in december 2001. Patients with rapidly progressive glomerulonephritis were excluded. Early (ER) and late referral (LR) were defined by the time of first nephrology encounter greater than or less than 6 months respectively, before iniciation of hemodialysis. RESULTS: 106 patients (76.25%) were referred early; mean follow-up time 6.3 +/- 4.5 years. 33 patients (23.74%) had late referral, follow-up time was less then six months, 18 patients were followed during less than 4 weeks. There were no differences in demographic data and comorbid conditions between LR and ER patients (age, cardiac and vascular disease, diabetes, neoplasia...). Mean plasma concentration of creatinine and urea was significantly greater, whereas hematocrit and albumin were less in the LRA than the ER group. Emergency dialysis through central vein catheterisation was more frequent in the LR group. Number of admissions and duration of hospital stay were higher in the LR group. No significant differences in nutrition, dialysis doses or anemia were found between the two groups after 6 and 12 months of hemodialysis. Long term outcome was similar in both groups: no significant differences were found in percentage of patients transplanted or deceased after 3 years of treatment. Survival analysis failed to show a difference between ER and LR groups (mean survival time was 73.6 +/- 4.3 months and 73.0 +/- 6 months respectively). CONCLUSION: Late referral to the nephrologist is associated with increased early morbidity vs early referral, although long term outcome is not worse if predialysis comorbid conditions are comparable and dialysis care achieve equal results in dialysis doses, nutrition and anemia in both groups in the first months of treatment. Improvement of outcome of patients referred early to the nephrologist will depend on the adoption of preventive measures over comorbidity factors that should be applied in early stages of renal disease. PMID- 12891940 TI - [Hypophosphatemia in dialysis units]. AB - Hypophosphatemia (Hf) is infrequently reported in chronic hemodialysis patients. The objective of this report is to describe the incidence, etiology, symptoms and treatment of Hf in a Dialysis Unit (defined as phosphorus < 2.5 mg/dL). In a retrospective study over a period of three years, we identified 22 cases of Hf, occurring on 11 among 149 patients. A two-groups distribution was made: Group A, patients with more than one episode (n = 3, 14 episodes of Hf) and Group B, patients with only one isolated episode of Hf (n = 8, 8 episodes of Hf). Plasma Ca, P, Albumin and nPCR were significant lower in group A (p < 0.05). Only two patients of group B had symptoms. Cases of Hf were: Group A: low-protein diet and alcoholism, Group B: decreased dietary intake due to non-digestive problems (n = 2) or due to digestive problems plus antacids (n = 4), phos-phate binders (n = 1) and dietary phosphorus restriction (n = 1). Three patients had secondary hyperparathyroidism. Treatment consisted on oral supplementarion by diet and changes in oral calcium salts. Intravenous supplementation was required acutely to raise serum P in a patient with auricular fibrilation. Two group A patients who has plasma 1.25 vitamin D < 5 pg/mL received vitamin D, and the third oral supplements of P. In all the cases, Hf resolved with these measures. We concluded that Hf is not so infrequent in hemodialysis. In patients with low-protein diet and low vitamin D concentration, Hf can be sustained. On the other hand, a decreased dietary intake maintaining similar phosphate binder's supplementation is the most frequent cause of occasional and symptomatic Hf, even in patients with secondary hyperparathyroidism. PMID- 12891939 TI - [Influence of different pharmacological agents in the ex vivo proliferation of mesothelial cells obtained from the peritoneal effluent of patients treated with peritoneal dialysis]. AB - Mesothelial cells (MC) are the first peritoneal membrane barrier in contact with dialysate. The aim of this study was to analyze the in vitro capacity of different pharmacological agents to modify the ex vivo proliferation of MC obtained from the peritoneal effluent of patients treated with peritoneal dialysis (PD). MATERIAL AND METHODS: Thirty cultures of MC taken from nocturnal peritoneal effluent were performed. After identification, MC are subcultured in 24 multi-well plates, adding the different exogenous agents. Proliferative capacity and cell morphology were estimated on day 16th of culture. The agents evaluated were insulin, IGF-1, tamoxifen, labetalol, carvedilol, enalapril and losartan. RESULTS: Insulin shows a dose-dependent effect on MC growth, with a limit that is stimulated by the addition of fetal bovine serum (FBS). Concentrations higher than 100 micrograms/ml, are not associated with further growth, even with cell damage. In contrast, the wide range of IGF-1 dose used did not affect to MC proliferation. Tamoxifen causes negative effects on MC growth just a very high doses, not resembling doses in clinical practice. Labetalol does not modify MC proliferation used under therapeutic calculated range. However, concentrations higher than 40 micrograms/ml showed a negative influence on growth, behaving as lethal doses that over 100 micrograms/ml. The addition of FBS attenuates this effect. These effects were very similar to that caused by carvedilol addition. Enalapril and losartan act as antiproliferative agents for MC. This effect is potentiated with angiotensin II, reaching lethal concentrations increasing the dose. In conclusion, mesothelial cell growth ex vivo taken from nocturnal peritoneal effluent on PD patients is an useful tool to explore the effects of any pharmacological agent on the biology of the cell of the peritoneum. The agents used had any influence in the proliferation capacity of mesothelial cells. PMID- 12891941 TI - [Ischemic colitis after renal transplantation:etiology and pathogenesis]. AB - Ischemic colitis is a well-recognized complication occurring in renal transplant recipients. It has often been associated with cytomegalovirus (CMV) vasculitis. However, the diagnosis of this pathology in the absence of CMV suggests that other etiological factors might be involved. Drugs inducing mesenteric vasoconstriction, such as non-steroidal anti-inflamatory drugs (NSAIDs) and cyclosporine could be related to this entity. PMID- 12891942 TI - [The spectrum of cystic kidney disease in adulthood: differential diagnosis and complications]. AB - Simple renal cysts are the most common renal masses, accounting for roughly 65 to 70% of cases. They most often occur in patients over the age of 50 as determined from post-mortem examination or renal ultrasonography. The major concern with simple renal cysts is differentiating them from more serious disorders, such as polycystic kidney disease and solid masses such as a renal carcinoma or abscess. Renal arteriovenous malformations may present with ultrasound picture mimicking simple parapelvic cyst. Ultrasound, doppler ultrasound, computed tomography and magnetic resonance imaging are effective in documenting the underlying lesions non-invasively. Arteriography may be useful to characterise vascular lesion. We report here the spectrum of cystic kidney disease in adulthood in a group of patient with different disorders. The differential diagnosis, complications and associated process are discussed. PMID- 12891943 TI - [Pure red blood cell aplasia associated with neutralizing antibodies against erythropoietin induced by epoetin alfa:a new form of acquired erythroblastopenia in auremic patients]. AB - We report the case of a woman with end-stage renal disease on maintenance hemodialysis and treated with recombinant human erythropoietin (epoetin alfa) administered subcutaneously, who developed a pure red cell aplasia secondary to the development of neutralizing antibodies anti-erythropoietin after 8 months of treatment. Despite epoetin withdrawal and immunosuppressive treatment with corticosteroids and gammaglobulins the patient still has high red blood cell transfusion requirements and undetectable plasma erythropoietin levels. Pure red cell aplasia secondary to the development of neutralizing antibodies anti erythropoietin is a rare but severe complication associated with the use of recombinant human erythropoietin in uremic patients. In recent years, the incidence of this complication has sharply increased, specially associated with the use of epoetin alfa administered subcutaneously. For this reason, the Spanish Drug Agency has recently contraindicated treating uremic patients with epoetin alfa administered subcutaneously. PMID- 12891944 TI - [Age and risk of mortality in renal transplanted]. PMID- 12891945 TI - [Purpura de Schonlein-Henoch in patient with fibrosis qistica]. PMID- 12891946 TI - [Nephrotoxicidad for imatinib]. PMID- 12891947 TI - CMS move threatens to put orphan drugs out in cold. PMID- 12891948 TI - 'Have I got a magic lamp for you!'. PMID- 12891949 TI - Compensation monitor. Where physician executives can expect the best pay. PMID- 12891950 TI - Predictive modeling, sharp lens on near future. PMID- 12891951 TI - A little something for the physicians. PMID- 12891952 TI - Money pit: is accreditation always worth the cost? PMID- 12891953 TI - A conversation with Gail R. Wilensky, PhD. Adviser to Presidents considers politics and reform. Interview by Patrick Mullen. PMID- 12891954 TI - Managing perinatal outcomes: the clinical benefit and cost-effectiveness of pharmacologic treatment of recurrent preterm labor. AB - PURPOSE: To compare the clinical benefit and cost-effectiveness of utilizing continuous subcutaneous terbutaline versus oral tocolytics following recurrent preterm labor. DESIGN: Retrospective, 1:1 matched cohort. METHODOLOGY: From prospectively collected data in a nationwide, perinatal database of women receiving outpatient services, we identified singleton gestations having recurrent preterm labor, stabilized during hospitalization, and subsequently treated with oral tocolytics (PO group) or continuous subcutaneous terbutaline infusion (SQ group). Those without medically indicated delivery were eligible for inclusion. Each woman in the PO group was matched 1:1 by gestational age at recurrent preterm labor to a woman in the SQ group. A standardized cost model was applied to compare total antepartum hospital, nursery, and outpatient charges. Wilcoxon Signed Rank, paired t, and McNemar's C2 test statistics were used for comparisons. PRINCIPAL FINDINGS: 558 women were studied (279 per group). The PO group had less gestational gain following recurrent preterm labor than the SQ group (28.4 +/- 19.8 days vs. 33.9 +/- 19.0 days, respectively, P < .001). The SQ group had less per patient charges ($) for antepartum hospitalization (3,986 +/- 6,895 vs. 5,495 +/- 7,131, P = .009), and nursery (7,143 +/- 20,048 vs. 15,050 +/ 32,648, P < .001). Outpatient charges were less for the PO group (1,390 +/- 1,152 vs. 5,520 +/- 3,292, P < .001). Overall costs for those in the SQ group were $5,286 less per pregnancy compared to the PO group. CONCLUSION: In this population, continuous subcutaneous terbutaline infusion was both a clinically beneficial and cost-effective treatment following recurrent preterm labor. PMID- 12891955 TI - Drug-eluting stents to transform cardiovascular medicine. PMID- 12891956 TI - HMO players continue earnings roll: how long will employers put up with it? PMID- 12891957 TI - National drug bill to continue rising by 14%-18%. PMID- 12891958 TI - Service enhancement to a dual-diagnosis population: mental illness/substance abuse (MISA). AB - This study details how the Sinnissippi Center, Inc., identified, eliminated, or modified barriers to accessing services for MISA (mentally ill and substance abusing) consumers to improve their level of functioning. Barriers and impediments to MISA consumers receiving the full range of services were identified; specific systems changes and modification and clinician help were implemented; and process measures, functional measures, and comparison measures were taken, as was fidelity to the improved service model. Comparison measures showed a significantly lowered dropout rate (5.5% versus 65%) for the MISA specific group and improved functioning over the comparison group. Process measures showed a significantly lowered use of emergency services, reduced emergency psychiatric hospitalizations, increased sobriety, decreased global assessment of functioning ranges, and an increased length of service. Functional measures showed a 199% increase in the 11 functional areas when averaged. Fidelity testing showed an average of 72% fidelity to the revised MISA services model. This study concludes that improved access to MISA-specific services significantly increases these consumers' quality of life. PMID- 12891959 TI - The GDAHA hospital performance reports project: a successful community-based quality improvement initiative. AB - During the past decade there has been increasing distribution of hospital performance information but few examples of how this information is affecting the quality of health care delivery. This article describes the methods of implementation and factors influencing a successful community-based quality improvement initiative in Dayton, Ohio, involving a collaborative of five competing hospitals in partnership with the business community and local and state hospital associations. The initiative contributed to a 36% reduction in acute myocardial infarction mortality over a 3-year period by changing reperfusion patterns in patients with ST segment elevated myocardial infarction. Identification of an opportunity gap, root cause analysis, and development of process measures used to facilitate health care provider change are summarized. The driving and restraining forces that have shaped this initiative from a report card to a quality improvement program are outlined and a list of five contributors to success are presented. These factors can serve as a basis for how other communities can benefit from this collaborative model. PMID- 12891960 TI - Quality improvement and the need for IRB review. AB - Improving health care quality is increasingly recognized as a national priority in the United States. As a result, more and more health care organizations can be expected to undertake quality improvement (QI) initiatives. A question being raised with increasing frequency is: "Which QI activities need review by an institutional review board (IRB)?" Structured data collection and analysis is a common characteristic of most QI activities. For some QI projects, the fundamental goal is improved understanding of phenomena presumed to be generalizable to settings other than those directly studied. These activities are research. For other projects, the fundamental goal is improvement in specific processes and systems within specific organizations. These activities are not research. This article proposes that this difference in intent and the fundamental nature of the activity is crucial in deciding which QI initiatives need IRB review. The article presents test questions and markers to distinguish research from other types of QI activities. Those that are not research do not require IRB review. However, because such activities may still put patients at risk, some other review may be necessary. The article proposes five levels of risk and makes recommendations for review by an entity other than the IRB. PMID- 12891961 TI - The coming third health care revolution: personal empowerment. AB - The financing and organization of health care in the United States has been rapidly evolving in the last 30 years. Managed care and capitation have largely replaced fee-for-service as a way to pay providers. Cost-control initiatives were developed by payers. These crude cost-control methods proved insufficient and new approaches were developed. These might be referred to as disease management that includes evidence-based medicine and outcomes measurement. It is proposed that a third revolution, patient empowerment, is just starting. The potential far reaching consequences are described, discussed, and analyzed here, including their cost consequences. PMID- 12891962 TI - Patient satisfaction with nursing care: a multilevel analysis. AB - Although prior research has suggested that satisfaction with nursing care is affected by multilevel factors (e.g., patient characteristics, episode-of-care, the institution providing care), these studies typically focused only on a single level of analysis. The present study examines three levels of influence simultaneously to assess the relative effect each has on satisfaction. Results suggest that satisfaction is determined primarily by the patient and the episode of care; organization-level factors explained almost no additional variance. PMID- 12891964 TI - [Practical evidence-based medicine]. PMID- 12891963 TI - QMHC interview. Interview with Thomas C. Shields, JD. PMID- 12891965 TI - [Evidence-based treatment and education programs--evaluation of complex interventions]. AB - Patient education and treatment programmes are complex interventions that require a multiphased stepwise evaluation. The continuum of increasing evidence of effectiveness of structured treatment and teaching programmes for diabetes and hypertension as implemented in Germany is presented, and the indispensable components of the program and the conditions of their effective implementation are described. PMID- 12891966 TI - [Evidence-based medicine as a model of decision making in clinical practice]. AB - INTRODUCTION: The way how we apply the principles of evidence-based medicine in the real clinical world is based on a number of implicitly assumed theoretical models. The aim of the present article is to identify these theories and use their impact to explain their importance for the application of evidence-based medicine. METHODS: Philosophical theories were analysed with regard to their impact on the present investigation, i.e. their ability to explain the implicit theoretical background to this way of practising evidence-based medicine. In a second step, the theories identified were used to discuss the question of clinical decision making. The model of the hermeneutic approach to clinical casework was used to introduce a strategy for everyday application of evidence based medicine in the clinical field. RESULTS AND CONCLUSION: In the context of these theories, there are three archetypal models of clinical decision making. These include the intuitive, the hypothesis-based and the evidence-based approach. In this sense, evidence-based medicine is a hermeneutic model for making clinical decisions. The mean impact of evidence-based medicine is the introduction of the best external evidence as a basis of the hermeneutic clinical casework. In the model of the "best external evidence", the clinician experiences a great change in clinical decision making. This change is generated by crossing the border from applying scientific clinical hypotheses to systematically identified and reviewed results of high-quality research work. In this sense, hermeneutic casework demonstrates that there is a contradictory unity between evidence-based clinical data and the clinician's understanding of a clinical case -a contradiction that may only be solved by the clinician's internal evidence. PMID- 12891967 TI - [Evidence-based pharmacology in community and hospital pharmacies--a vision of the future?]. AB - Pharmacists are academically trained drug and medication specialists. Also in the eyes of the law, pharmacists are more than just specialised sales people for medication. As laid out in the regulations governing pharmacies, pharmacists see patients and experts in the fields of medicine, dentistry and veterinary science to whom they have to offer advice. The pharmacist's advisory activities, though, must not interfere with an existing doctor-patient relationship. All pharmaceutical activities in community and hospital pharmacies should therefore be based--similar to the medical faculty--on an evidence-based practice in the sense of an evidence-based pharmacy adopting all principles of Evidence-based Medicine. The challenge for pharmacists and doctors should therefore be to work together to increasingly develop common strategies in order to achieve optimal treatment for the benefit of the patient. Especially in the treatment of chronically ill patients, synergistic effects could be reached through open exchange and a closer linkage between medical and pharmaceutical activities. Such closer contacts between the two disciplines could be made at interdisciplinary colloquia or special subject-specific round table discussions. Pharmaceutical Care (PhC), i.e. the optimisation of medicational therapy of chronically ill patients through accompanying intensive pharmaceutical advice is desirable from the patient's point of view and could take some load off medical practitioners. Any questions as to the use of medication, potential side-effects or undesirable interaction with other medications, but also regarding additional preventive measures can be posed to qualified personnel by the patient collecting his/her medication in the pharmacy. Also, as community pharmacists see themselves as active medication specialists and valuable qualified advisors to the patient with regard of self-medication, increased investment in internal communication between colleagues and external communication with patients is desirable. In the hospital setting, the pharmacy is to be regarded as the central medication information office and should participate in interdisciplinary working groups for the development of clinical guidelines and substitution possibilities. Furthermore, hospital pharmacy should be consulted on pharmaco-economic questions and on planning and conducting clinical studies. PMID- 12891968 TI - [The practice of evidence-based medicine in surgery--evidence and experience]. AB - INTRODUCTION: The important role of technical expertise, experience and intuition in the operative specialties may hamper both clinical research and evidence-based medicine (EBM). We assessed whether the results of EBM practice projects actually varied between surgery and medicine. METHODS: EBM practice projects were identified through a search in Medline. We examined the proportion of questions, for which evidence was found and which led to a change in clinical management. At the same time, the proportion of randomised controlled trials (RCTs) in surgical (95) and general internal medicine (84) journals was compared. RESULTS: Of the 13 EBM practice projects identified, only two dealt with surgical patients. While the proportion of answerable questions was similarly high in all projects (61% to 96%), clinical impact was lower in surgical (9% and 18%) than in medical projects (20% to 48%). Compared to medical journals, surgical journals contained fewer RCTs (3.6% vs. 2.5%; p = 0.168). CONCLUSION: The clinical impact of EBM seems to be lower in the operative specialties. This is probably due to the slightly poorer quality of surgical studies or to the difficulties of applying research results in one's own practice. PMID- 12891969 TI - [Evidence-based medicine in the real world--an experimental evaluation in a hospital care system]. AB - In Germany, effective tools for the implementation of evidence-based medicine (EbM) in the hospital setting have been rarely recorded. In the model project of the Park-Klinik Weissensee in Berlin, the so-called Park-Klinik EbM project, a multi-dimensional integrational approach is being put to the test. The Berlin hospital regards recommendations for actions based on EbM in the form of text modules (EbM bits of information) employed for discharge letters of the Department of Internal Medicine and locally tailored clinical practice guidelines as appropriate means for the implementation and application of EbM. Here we present conceptual developments and the first results of quantitative and qualitative evaluation. The benefits of EbM efforts are critically (positively) appraised. The dissemination and implementation of clinical practice guidelines into everyday evidence-based practice must still be improved, though. EbM bits of information are successful and effective instruments for both clinicians and general practitioners. PMID- 12891970 TI - [An experimental evaluation of a hospital's internal clinical guideline system. Randomized, controlled crossover pilot study on the efficacy of Intranet-based guidelines for General Internal Medicine at the Park Clinic Weissenssee in Berlin]]. AB - Randomised, controlled crossover pilot study on the efficacy of Intranet-based guidelines for General Internal Medicine at the Park-Klinik Weissensee in Berlin. As part of the evaluation programme for the intranet-based internal guideline system at the Park-Klinik Weissensee in Berlin, an experimental randomised, controlled trial was conducted in which 12 medical students in their final year were asked to solve 10 clinical vignettes each. A number of written and electronic sources of medical information was available. In addition, the students were allowed to use the internal guideline system for one half of the vignettes in a crossover fashion. There was a sample size-related non-significant trend in favour of the guideline system concerning the primary endpoint "time to right solution" (mean 2.8 min; 95% CI -1.6 to 7.3). A per-protocol-analysis revealed a great variability in the actual use of the guidelines available. There was an inverse "dose-response relationship" between the intensity of guideline usage and the endpoint. PMID- 12891971 TI - [Pedagogic reflexions on the evidence-based medicine curriculum of the German Central Agency for Quality in Medicine and the German Network for Evidence-Based Medicine]. AB - The German Agency for Quality in Medicine (AQuMed) and the Network for Evidence based Medicine (EbM) have recently published the draft of an EbM Curriculum for continuing medical education. This standardised concept might lead to an improvement of the quality of EbM courses for physicians. Critical appraisal of the curriculum from a didactic point of view reveals both a substantial lack of teaching methods and teaching aids, and a lack of appropriate tools for the evaluation of the actual increase in professional competence. This includes specialised subject knowledge, methodological considerations as well as highly developed communication and social skills. We recommend incorporation of these components into the curriculum design to facilitate its successful implementation. PMID- 12891972 TI - [How many standards can variety take?]. AB - The recently published "Curriculum Evidence-based Medicine" of the German Network Evidence-based Medicine and the German Agency for Quality in Medicine has been criticised for shortcomings in teaching methods and the teaching process. In this article, authors of the curriculum provide a replique to the criticisms. PMID- 12891973 TI - [Evidence-based medicine in professional training and education in practice? The integrated evidence-based medicine curriculum at the Medical School at the University of Witten/Herdecke]]. AB - Applying the principles of evidence-based medicine in clinical practice is becoming more and more important for healthcare professionals in Germany. In this context Witten/Herdecke University has implemented an EbM Curriculum for medical students that is closely related to problem-based learning and clinical rotations within the scope of the six-year medical reform curriculum. Starting with the first semester, students will be able to acquire the necessary skills to search medical databases, to critically review the information retrieved according to EbM standards, to gather evidence for answering diagnostic or therapeutic questions during their clinical education and to ultimately participate in the development of clinical guidelines during their last year. PMID- 12891974 TI - [A new measure. New studies require new criteria for critical appraisal]. AB - Over the last ten years the quality of published clinical trials has improved considerably. Randomised controlled design, concealed allocation, blinding, clinically relevant outcomes, large sample sizes and almost complete follow-up are standard features of most trials. As a result, standard evidence-based medicine criteria can no longer be used to separate the "wheat from the chaff". Here, we discuss recent studies which reveal only marginal superiority of new treatments over well-established and effective therapies. Nowadays, readers of reports should be particularly aware of the definition of control therapies, of clinical relevance when results are statistically significant, of the external validity and whether a trial serves a pragmatic or explanatory purpose. PMID- 12891975 TI - [Evidence-based medicine in 2025--what will remain?]. AB - The reader is invited to attend a symposium on the long-term impact of Evidence based Medicine (EBM) held in 2025. When EBM emerged for the first time, there had been great expectations, not all of which were fulfilled. Against the background of many vested interests evidence from clinical studies was not always welcome. Nor had EBM been able to abolish controversy and conflict within the health care system. But EBM had been associated with a substantial change of clinical mentality with practitioners becoming more cooperative, more self-critical and open for a scientific basis of their work. Although the reasoning of doctors has remained within a narrative framework, EBM has provided them with a critical tool for diagnosis, causal relations and intervention. PMID- 12891976 TI - [Methods in evidence-based medicine consensus findings--a case study in trauma surgery]. AB - The methods and processes of a guideline development programme including a structured consensus conference for the employment of Diagnostic Imaging in Trauma Surgery are presented and critically reviewed. International standards and methodical concepts described in the Guideline Manual by the Agency for Quality in Medicine (AQuMed) and the Association of the Scientific Medical Societies in Germany (AWMF) have been implemented. Experiences with this project reveal that even a small multidisciplinary panel group is potentially able to cope with the comprehensive panel work if they are methodologically supported. The establishment of an internal quality management proved to be a useful tool for the production of high-quality results in the face of limited project resources. As expected, for most sub-questions knowledge of evidence and medical experience correlated well. However, the lack of a method for ensuring that consensus recommendations finally obtained by panel work and discussions during the conference were always based on evidence was the weak spot of this evidence-based consensus conference. PMID- 12891977 TI - [German Society for General Medicine--guidelines for urinary incontinence: content and development]. AB - In this article, the authors of the "General Practitioners' Guideline to the Diagnosis and Treatment of Urinary Incontinence" describe the historical development of this evidence-based GPs' guideline for the management of urinary incontinence. The guideline's history is focused around the following points: 1. reasons for choosing the subject matter, 2. material and personnel expenses of the development process, 3. specific considerations regarding recommendations that will be applied to the primary care setting. The whole guideline project was based on the hypothesis that developing and implementing a clinical guideline for general practitioners in Germany would lead to better detection and treatment of cases of urinary incontinence. Hopefully, the guideline will contribute to a better quality of life of the persons affected and to a reduced number of "silent sufferers". Specifically, the targets to be addressed include the following: identifying and providing treatment to previously undiscovered sufferers of urinary incontinence implementing a universal standard of scientifically approved diagnostic strategies in family physician's offices enhancing the therapeutic options available to patients suffering from urinary incontinence in the community aiding primary care physicians with the diagnosis and treatment of people with urinary incontinence improving the interprofessional relationships with other carers in the field of urinary incontinence. The authors wish to thank the German Cochrane Centre and the Deutsche Krankenversicherung for organising the Cochrane Prize and thus promoting guideline development. PMID- 12891978 TI - [Value of evidence-based medicine in education and continuing education]. PMID- 12891979 TI - [Integrating problem-based learning into traditional medical courses: the PBL approach to Pediatrics in Berlin]. AB - Integrating a problem-based learning (PBL) approach into the classical curriculum for medical students is often considered difficult because of fundamental differences in the two teaching methods. At the Charite, the medical school of Berlin's Humboldt University, students with a regular medical curriculum are offered to choose between a regular and a PBL class for their course in Paediatrics. During the three-week course in PBL Paediatrics medical students spend the morning in different clinics of the "Otto Heubner Centre for Paediatrics". They are strongly involved in the medical care of 3-4 patients with typical paediatric diseases. Each afternoon, one of the students presents a patient. Patient problems are discussed according to the seven steps of PBL. Subsequently, students use different tools and sources through self-studies to obtain information on the specific disease. By integrating the principles of "Evidence-based medicine" students are trained to critically appraise the medical literature. At the end of the course students are asked to evaluate the quality of the medical teacher, thus allowing for continuous improvement of the teaching methods. We conclude that the principles of PBL can be integrated into a conventional curriculum in medical school. PMID- 12891980 TI - [Change in the nursing process: in 5 work relevant steps to nursing practice]. PMID- 12891981 TI - [Physical therapy in obstructive respiratory tract diseases: when dyspnea increases]. PMID- 12891982 TI - [Pneumonia and atelectasis prevention in general practice: efficiency comes with engagement]. PMID- 12891983 TI - [Noninvasive positive pressure ventilation (NIPPV): many advantages, but not without risk]. PMID- 12891984 TI - [Nursing care of patients with thoracic drainage: safe respiration]. PMID- 12891985 TI - [Nursing patients with mucoviscidosis: living with a limited future]. PMID- 12891986 TI - [Treatment pathways for chronically ill patients--: Holistic support by theoretical background]. PMID- 12891987 TI - [New nursing regulation: a chance for the future?]. PMID- 12891988 TI - [What can a professional nursing association accomplish? "From nothing comes nothing"]. PMID- 12891989 TI - [Concept for training new coworkers in artificial ventilation: knowledge brings security]. PMID- 12891990 TI - [Nicotine withdrawal in hospital coworkers: opportunities instead of prohibitions]. PMID- 12891991 TI - [Counseling in the service center for respiratory therapies: competence provides trust]. PMID- 12891992 TI - [Personal stories: I want to be your pain]. PMID- 12891993 TI - [Evidence-based nursing educational center in Bologna: research to improve patient outcome]. PMID- 12891994 TI - [From case to case: idleness is the source of all troubles]. PMID- 12891995 TI - [Independent nursing career: "Flexibility is necessary"]. PMID- 12891996 TI - [Wound care, 7--Patients with temporary tracheostomy: knowledge and careful nursing are essential]. PMID- 12891997 TI - [Prevention of decubitus ulcer: what is taught, what will be done, what is evident?]. PMID- 12891998 TI - Treatment of myoclonus. AB - Myoclonus is a clinical term meaning a sudden, quick, involuntary muscle jerk, irregular or rhythmic, arising in the central nervous system. The most important initial step in the treatment of myoclonus is to try to subclassify the type of myoclonus and identify the underlying disease process. Metabolic derangements or other underlying conditions, if found, need to be treated. Offending drugs causing myoclonus should be removed. A number of different drugs have been used for the symptomatic control of myoclonus. They include sodium valproate, clonazepam, some other antiepileptic drugs, piracetam, and levetiracetam. PMID- 12891999 TI - Cost of acute stroke. A review. AB - The annual incidence of cerebrovascular diseases in Belgium is between 200 and 230/100,000 inhabitants. Mortality after stroke is about 21% and approximately 30% of stroke patients will be dependent on others. Cost is mainly related to the length of hospital stay but also to outpatient care. Length of stay is dependent on stroke severity at entry, stroke location (total infarct in the anterior circulation costs twice as much as small lacunar infarcts), and the social status (patients living alone or in a nursing home have a longer length of stay gene rating a higher cost). Stroke units by using rationalized acute stroke therapy (including thrombolysis) reduce death and disability and decrease the length of stay by acting on stroke severity and co-morbidities. Stroke units also facilitate early discharge by coordinating outpatient care with the general practitioners, rehab centers, and nursing home. A global stroke-based prospective payment should be implemented in Belgium. PMID- 12892000 TI - Anti-epileptogenesis research: the clinical relevance. AB - In recent years, different research lines have examined the epileptogenic process in order to understand the different stages in this process, and with the hope that early recognition and intervention could prevent chronic epilepsy in patients with epileptic seizures. In animals, acquired epilepsy is studied most commonly with kindling models, status epilepticus models and traumatic brain injury models. Molecular genetic studies substantially help to understand age specific channel and receptor abnormalities. Major progress has been made in recent years and we are now waiting for the first large scale multi-center clinical trials that test the possible anti-epileptogenic properties of anti epileptic drugs or other compounds in well defined patient groups. In clinical practice, a structured diagnostic work-up in all patients with recurrent seizures is a first and necessary step in the recognition of patients at risk for developing chronic and refractory epilepsy. PMID- 12892001 TI - Abnormal psychophysical visual perception in Parkinson's disease patients. AB - Several visual dysfunctions in Parkinson's disease (PD) are described. Most of them are subtle or only demonstrated by stimulus-specific electrophysiologic or psychophysical testing. However, these minor deficits are thought to be of clinical relevance as they are related to direct or indirect complaints. Special emphasis is laid on visual hallucinations. These are most likely of multifactorial origin. The relation between hallucinations in PD and in dementia with Lewy bodies has to be elaborated further. Visual loss, as a possible and reversible cause of visual hallucinations should be actively sought and corrected as far as possible. An underlying role of dopaminergic retinal cells in visual dysfunction of PD patients is widely recognised. However, whether the basic abnormality resides also in the visual cortex remains to be elucidated. Other neurotransmitters may also be involved. It has not been answered whether visual dysfunction might distinguish PD from other forms of parkinsonism. PMID- 12892002 TI - De novo cryptogenic refractory multifocal febrile status epilepticus in the young adult: a review of six cases. AB - BACKGROUND: Status epilepticus (SE) is a common neurological emergency. In around 10 percent, the etiology of SE is not clear, i.e. cryptogenic SE. AIM: To describe six young adult patients with de novo cryptogenic multifocal febrile SE with poor outcome. METHODS: Retrospective chart review. RESULTS: We describe six patients who presented with de novo SE. All were young adults with an unremarkable previous medical history. In all except one, the family history for epilepsy was negative. All had a febrile illness in the week preceding SE. Investigations were unable to reveal the underlying etiology. SE proved refractory to conventional antiepileptic drug treatment (AED) and anesthesia was required to control seizures. Seizures during SE or ensuing epilepsy were multifocal. Brain pathology in four patients was normal in two and showed changes secondary to seizures in two. Outcome was poor in all cases: one patient died during the course of SE, another patient remained with severe mental and physical disability as well as refractory epileptic seizures, the other four developed refractory multifocal epilepsy. One of these patients died a sudden unexpected death in epilepsy (SUDEP) and one drowned during a seizure. CONCLUSIONS: Six patients were reviewed who presented with de novo cryptogenic multifocal refractory febrile SE. The possible etiologies are discussed. Based on the similarities in these cases, a distinct refractory SE syndrome might be assumed. Further prospective studies will, however, be required to validate this hypothesis as well as to elucidate its etiology and optimize treatment. PMID- 12892003 TI - Suspected lamotrigine-induced toxic epidermal necrolysis. AB - Toxic epidermal necrolysis and Stevens-Johnson syndrome are rare, life treating cutaneous reactions. Most cases of toxic epidermal necrolysis are drug induced. The drugs with the highest estimated incidence include co-trimoxazloe (trimethoprim-sulfamethoxazole), sulfadoxine-pyrethamine, and carbamazepine. Among other drugs, the reported reaction rates are relatively low for lamotrigine and sulbactam-ampicillin. We describe a patient who developed toxic epidermal necrolysis after either administration of lamotrigine or of ampicillin. PMID- 12892004 TI - Recurrent transient ischemic attacks in a 15-year-old boy with beta-thalassemia minor and thrombophilia. Contribution of perfusion SPECT to clinical diagnosis. AB - beta-Thalassemic patients exhibit an increased frequency of thrombotic events but most patients with heterozygous beta-thalassemia minor are asymptomatic and no single case with beta-thalassemia minor and concurrent stroke was reported. We present a 15-year-old boy with heterozygous beta-thalassemia minor who developed recurrent transient ischemic attacks as documented with repeated brain SPECTs whereas structural neuro-imaging was not contributory. The patient exhibited resistance to activated protein C due to heterozygosity for factor V Leiden as well as slightly decreased plasma levels of protein C and S. This unique association of risk factors might have caused clinically significant thrombophilia resulting in recurrent cerebrovascular events. This case report underlines the thrombogenic risk of heterozygous beta-thalassemia minor in children heterozygous for factor V Leiden mutation. We therefore suggest to screen for thrombophilia in children with beta-thalassemia minor when thromboembolism-related phenomena occur. This case also demonstrates that brain perfusion SPECT is a useful and sensitive tool for detecting cerebrovascular events in patients with hemoglobinopathies. PMID- 12892005 TI - Acquired cerebellar cavernous angioma following childhood radiotherapy in a patient with neurofibromatosis type 1. AB - We report the unusual case of a patient with neurofibromatosis type I, who was irradiated 16 years previously for a mesencephalic glioma causing hydrocephalus, and who developed a cerebellar haemorrhage caused by a histologically confirmed cavernous angioma, that was invisible on several earlier MRI scans. The different hypotheses concerning the de novo formation of cavernous angiomas (venous obstructive disease and genetic abnormalities) are succinctly reviewed. PMID- 12892006 TI - [Two patients with fever, chills and pain in the right upper abdominal quadrant accompanied by hyperbilirubinaemia: pylephlebitis]. AB - Two male patients aged 55 and 77 years, respectively, presented to the casualty department with fever, chills and right abdominal upper quadrant tenderness. They also had hyperbilirubinaemia. Based on CT scan findings and blood cultures yielding Bacteroides fragilis, a diagnosis of pylephlebitis (septic thrombophlebitis of the mesenteric veins and/or the portal vein) was made. This is a condition with a mortality rate of 10-70%. Primary sources such as diverticulitis are often seen in patients with pylephlebitis, in which bacteria are drained by the mesenteric veins and cause a thrombus in the portal system. In the two patients no primary focus was detected. They were treated with intravenous antibiotic therapy followed by oral antibiotics, and were discharged in good health. Pylephlebitis can be complicated by liver abscesses. Treatment consists of broad-spectrum antibiotics which are adjusted based on the blood cultures results. The duration of treatment is between two and six weeks, depending on the presence of liver abscesses. In patients with abscesses that cannot be drained, longer treatment may be indicated. PMID- 12892007 TI - [Comments on the Dutch Institute for Health Care Improvement's standard code of practice entitled, "Donation of fetal tissue"]. AB - Under the auspices of the Kwaliteitsinstituut voor de Gezondheidszorg CBO [Dutch Institute for Healthcare Improvement] a standard code of practice was developed as a template for local institutional codes to implement the Law on Foetal Tissue. It is a useful model code, but arguments should have been outlined more explicitly, notably in instances where the code adopts a somewhat stricter position than the Law. The following remarks pertain to the model code: 1. It may be argued that the inclusion or exclusion of 12-15-year-old pregnant girls should be relative to the privacy-related sensitivity of the use of foetal tissue. 2. Transplantation requires additional tests for the safety of the recipient, including testing for HIV/AIDS. A pregnant woman's permission for such testing should not be taken for granted. 3. The abortion technique may be modified in view of the subsequent use of foetal tissue if the woman consents to the modification, with the prerequisites that the modification does not harm the woman and that any potential sensation of pain by the foetus must be minimised. PMID- 12892008 TI - [Nerve sparing radical hysterectomy in the case of cervical cancer]. AB - Autonomous pelvic nerves are essential for bladder and rectum function as well as sexuality. These nerves are usually permanent damaged during radical oncological surgery and this results in urological, sexual and proctological morbidity. Japanese surgeons have paved the way for surgical approaches to dissect these nerves during surgery and leave them intact. At the Department of Gynaecology of the Leiden University Medical Center, the Netherlands, a new surgical technique has been developed to spare the autonomous pelvic nerves during radical hysterectomy. In this procedure the lateral parts of the sacro-uterine ligament, through which the N. hypogastricus passes bilaterally, are left intact. The initial results of this nerve-sparing surgical technique during radical hysterectomy seem to be promising. The operation was easy to perform, with almost no extra blood loss and an extra operating time of just 10-15 minutes. Postoperative recovery of the bladder function occurred after a mean period of 9 days. PMID- 12892009 TI - [The effects of physical exercise on the immune system]. AB - Physical exercise has numerous effects on the human body, including the immune system. After strenuous exercise, athletes pass through a period of impaired immune resistance. During this period, athletes are theoretically more susceptible to upper respiratory tract infections, although a causal relation has never been demonstrated. Moderate exercise seems to have a beneficial effect on the immune function, which could protect against upper respiratory tract infections. Exercise has effects on both the humoral and the cellular immune system. Doping products, except glucocorticoids, only have modest effects on the immune system, although erythropoietin may, in rare cases, cause severe side effects. Glutamine and vitamin C could, hypothetically, prevent the negative effects of strenuous exercise on the immune function, but further studies are needed to demonstrate and explain these effects. PMID- 12892010 TI - [Dutch Institute for Health Care Improvement's standard code of practice entitled, "Donation of fetal tissue"]. AB - The Dutch Law on Foetal Tissue (Wet Foetaal Weefsel) sets out conditions and regulations concerning the donation, storage and permissibility of use of foetal tissue. Each institution where foetal tissue becomes available has to formulate an in-house code of conduct describing how the law will be applied within that institution. A committee, including abortion physicians and gynaecologists, has formulated a number of rules of conduct within the standard code of conduct under the auspices of the Kwaliteitsinstituut voor de Gezondheidszorg CBO [Dutch Institute for Healthcare Improvement]. Complying with this standard code ensures that the law is upheld. In clinical practice, one may deviate from this standard code in case of solid reasons only. Generally, all women can be asked to donate foetal tissue. A basic prerequisite for the inclusion of non-Dutch women is that oral and written communication is possible. The committee considers the age of 16 as a minimum threshold and mental incompetence as an exclusion criterion. Much attention should be paid to providing the women with information and to requesting permission. The basic principle is that the decision to terminate a pregnancy should be strictly separated from the decision to donate foetal tissue. As a general rule, foetal tissue should be stored anonymously. Adjustment of the abortion method in view of the intended scientific research is not permitted. PMID- 12892011 TI - [Diagnostic image (147) A man with gustatory sweating. Frey's syndrome]. AB - A 73-year-old man suffered from gustatory sweating after submandibular gland removal. Frey's syndrome was diagnosed and successfully treated with intracutaneous injections with botulinum A toxin. PMID- 12892012 TI - [A diagnostic decision rule for managing children with signs of meningeal irritation]. AB - OBJECTIVE: To develop a diagnostic decision rule based on clinical features to predict the risk of bacterial meningitis in children with signs of meningeal irritation. DESIGN: Retrospective and prospective. METHOD: Predictors for bacterial meningitis were identified by collecting clinical data from the records of 360 patients (aged 1 month to 15 years) who consulted the Casualty Department, Sophia Children's Hospital, Rotterdam, the Netherlands, with signs of meningeal irritation during the period 1988-1998. The diagnostic decision rule derived was prospectively validated on 226 similar children who consulted the casualty departments of four hospitals in the Netherlands during the period 1999-2001. RESULTS: Predictors for bacterial meningitis were the main complaint and vomiting (in the history) persisting for a long time, the presence of meningeal irritation, cyanosis, petechiae, disturbed consciousness (during physical examination), and a high serum concentration of C-reactive protein. Liquor analysis parameters with an added diagnostic value were the total polymorphonuclear cell count in the liquor and the liquor/blood glucose ratio. The use of these patient characteristics in a decision rule accurately predicted the chance of bacterial meningitis. The rule can also be used to refine the indication for lumbar puncture and empirical antibiotic treatment. PMID- 12892013 TI - [Prevalence of diabetes mellitus and cardiovascular disease among immigrants from Turkey and Morocco and the indigenous Dutch population]. AB - OBJECTIVE: To gain insight into the prevalence of diabetes mellitus and cardiovascular disease among Turkish and Moroccan immigrants in the Netherlands. DESIGN: Secondary analysis of data from a cross-sectional study in 1999/'00. METHOD: The prevalence of self-reported diabetes and cardiovascular disease was studied in a sample of 743 Turkish, 641 Moroccan and 537 ethnic Dutch persons aged 35 to 74 years from the population of Amsterdam, the Netherlands, using an oral questionnaire. Ethnic differences in the prevalence of diabetes and cardiovascular disease were studied by logistic regression analysis controlling for age, sex and educational level. Further, in the case of diabetes, we examined whether or not the differences could be explained by overweight and physical inactivity, and in the case of cardiovascular disease we investigated the relationship with smoking and diabetes. In addition, the association between diabetes and cardiovascular disease was investigated. RESULTS: The Turkish and Moroccan population reported diabetes more often than did the ethnic Dutch (12.3, 12.4 and 3.0%, respectively). The differences were still present after controlling for sex, age, educational level, overweight and physical inactivity. Cardiovascular disease was more prevalent among the Turks (10.6%) than among the indigenous Dutch population (5.0%), but this was not true for the Moroccans (5.4%). The difference between Turkish and ethnic Dutch people still persisted after controlling for educational level, overweight, physical inactivity, smoking and diabetes. The association between diabetes and cardiovascular disease was consistent for all groups studied. CONCLUSIONS: The prevalence of diabetes was higher among Turkish and Moroccan immigrants than among the indigenous Dutch population. Cardiovascular disease was more prevalent among the Turks, but not among the Moroccans. The association between diabetes and cardiovascular disease was consistent for all three ethnic groups. PMID- 12892014 TI - [Mycobacterium haemophilum as the cause of lymphadenitis in the neck in an otherwise healthy boy]. AB - In an 8.5-year-old boy with severe recurring unilateral swelling of the neck, infection with Mycobacterium haemophilum was established. In addition to the more usual causative agents, cervical lymphadenopathy in children can, in rare cases, be caused by M. haemophilum. The skin tests did not differentiate between Mycobacterium avium and Mycobacterium tuberculosis infection. There was no evidence of specific immune deficiencies. The swelling did not respond to treatment with four tuberculostatic drugs (rifampicin, isoniazid, pyrazinamide and ethambutole), nor to repeated surgical excision. Finally, M. haemophilum grew on the culture despite sub-optimal culture conditions. Thereupon the patient was treated with co-trimoxazole and recovered in two weeks. This is the first patient with cervical lymphadenopathy caused by M. haemophilum to be described in the Netherlands. M. haemophilum grows on an iron-rich medium at a relatively low temperature (30-32 degrees C). In cases of cervical lymphadenopathy, it is advisable to consider M. haemophilum as a cause and to adjust the culture conditions accordingly. PMID- 12892015 TI - [Withdrawal symptoms in a neonate following exposure to venlafaxine during pregnancy]. AB - Withdrawal symptoms occurred in a male neonate after maternal use of venlafaxine for depression during pregnancy. The symptoms were restlessness, hypertonia, jitteriness, irritability and poor feeding. The diagnosis was confirmed by a temporary improvement after administration of a low dose (1 mg) of venlafaxine to the boy. Eventually the symptoms began to decline spontaneously, and ceased after 8 days. Exposure to venlafaxine and other antidepressants which inhibit serotonin reuptake during the third trimester of pregnancy carries the risk of a neonatal withdrawal syndrome. PMID- 12892016 TI - [A case of fraud in a neurological pharmaceutical clinical trial]. AB - This paper describes the laborious and lengthy path to clarification and disclosure in a case of fraud in a neurological pharmaceutical clinical trial in the Netherlands. A Dutch neurologist was suspected of irregularities within the context of the 'European stroke prevention study 2' (ESPS-2), a multicentre study into medicinal prophylaxis in patients who had suffered a stroke. The Netherlands Society of Neurology (NVN) established an independent inquiry committee for further investigation of the case. The identity of 425 of the 438 patients (97%) included in the trial by the neurologist could be retrieved. The majority of these patients were known to the neurologist with cerebral infarct. For a sample of 115 patients, the general practitioners (GPs) were contacted by means of a questionnaire. Ninety percent of the responding GPs were unaware of their patients' participation in the pharmaceutical clinical trial. A total of forty patients were asked by their GP about participation: 36 (90%; 95%-CI: 76-97) indicated that they had not participated in the trial, and 4 could not remember. The committee concluded that the neurologist had committed fraud, in the sense that he had used the names of existing patients without these patients actually being enrolled in the study. The report of the independent committee was not made public; the committee and the NVN board differed in opinion on the interpretation and implications of the agreements regarding this subject. Following prolonged legal action, the regional Disciplinary Board suspended the neurologist from practice for one year and the court of law sentenced him to 180 days imprisonment or a fee of 130,000 Euro. Based on the experience gained from this case, recommendations in case of suspicion of fraud are discussed, such as the timely appointment of an independent inquiry committee and the establishment of unambiguous agreements regarding the disclosure of the results of the investigation. Possible legal implications should be considered in advance by the organisations involved; statutes should provide regulations for procedural rules. In the Netherlands there now exists a National Body for Scientific Integrity and a committee for the Scientific Integrity of Healthcare Research to prevent scientific misconduct and to stimulate reporting and appropriate handling of this problem. PMID- 12892017 TI - [A man with Campylobacter endocarditis, treatable as Campylobacter fetus following identification]. PMID- 12892018 TI - [Surgical treatment of tumor metastases in the lungs, brain or liver]. PMID- 12892019 TI - [Complications following liposuction]. PMID- 12892020 TI - [From gene to disease; Wilson disease: copper storage due to mutations in ATP7B]. PMID- 12892021 TI - [The value of the pulmonary-artery catheter: not ruled out, but not proven either]. PMID- 12892022 TI - JCAHO unveils medication-management standards. PMID- 12892023 TI - Pharmacists play vital role on USNS comfort. PMID- 12892024 TI - WHI findings another blow to hormone replacement therapy. PMID- 12892025 TI - The human factor in medication-error reduction. PMID- 12892026 TI - Risks and benefits of long-term hormone replacement therapy. AB - Studies of long-term hormone replacement therapy (HRT) are reviewed, and treatment recommendations based on the results are advanced. HRT is a popular therapy for postmenopausal symptoms and osteoporosis prevention in women. However, women also use hormones for unlabeled indications, such as cardiovascular disease prevention. In the past, observational and case-controlled trials have suggested that HRT confers a benefit through an improvement in lipid profiles and has relatively few adverse effects. However, no randomized controlled trial had proven this before HRT became popular. More recently, the results of additional observational studies and large, double-blind, placebo controlled trials have been published that indicate that HRT may not be as beneficial or risk free as first thought. If a woman wishes to begin or continue HRT for short-term menopausal symptoms, it is crucial to evaluate her individual risk of breast cancer, coronary heart disease, venous thromboembolism, and stroke before recommending therapy. Otherwise, HRT should not be recommended for treatment durations of more than five years, and treatment should be discontinued in women at risk of complications. Recent large, randomized, placebo-controlled trials have shown substantial risks and limited benefits in the long-term use of HRT. PMID- 12892027 TI - Alfuzosin hydrochloride for the treatment of benign prostatic hyperplasia. AB - The chemistry, pharmacology, pharmacokinetics, clinical efficacy, and adverse effects of alfuzosin hydrochloride in the treatment of benign prostatic hyperplasia (BPH) are discussed. Alfuzosin is a functionally uroselective alpha 1 adrenergic antagonist indicated for the management of moderate to severe BPH. It can improve urinary voiding symptoms and increase urinary flow rates while causing few cardiovascular adverse effects. When administered as an immediate release (IR) formulation, alfuzosin must be administered twice or thrice daily. The extended-release (ER) formulations of alfuzosin for once- or twice-daily administration are associated with small variations in peak and trough serum drug levels, which may contribute to the lower frequency of cardiovascular adverse effects reported with ER versus IR alfuzosin. Alfuzosin has been shown to improve patients' perception of quality of life, allowing patients to increase their physical activities and improve their ability to handle day-to-day activities. Less significant improvements in patients' sense of well-being and improved sexual functioning have been reported. The usual dose of alfuzosin for patients with BPH is 2.5 mg twice or thrice daily of the IR formulation or 5 mg of ER alfuzosin twice daily or 10 mg of ER alfuzosin once daily. The Food and Drug Administration is currently reviewing the ER 10-mg formulation for once-daily administration. IR alfuzosin is similar to all other second-generation alpha 1 adrenergic antagonists in mechanism of action, clinical efficacy, and adverse effects. No dosage titration is needed for ER alfuzosin, and its onset of peak action is within days of the start of treatment. PMID- 12892028 TI - Enterobacter cloacae bloodstream infections in pediatric patients traced to a hospital pharmacy. AB - The sources of an outbreak of Enterobacter cloacae bloodstream infections in a pediatric hospital were investigated, as were the risk factors for acquiring the infection: Two retrospective case-control studies were conducted. The study sample included all patients admitted to the general pediatric wards from February 5 through March 30, 2001, who had a positive blood culture for E. cloacae. Pediatric ward and pharmacy infection-control practices were reviewed, personnel and environmental cultures were obtained, and pulsed-field gel electrophoresis (PFGE) molecular typing of the bloodstream isolates was conducted. Four subjects were identified. These infants were more likely than control patients to receive i.v. ranitidine (p < 0.01). Among patients receiving i.v. ranitidine, subjects were more likely than controls to receive i.v. ranitidine prepared by a pharmacist. No environmental or personnel cultures yielded E. cloacae. Patients' E. cloacae isolates had four different PFGE patterns, suggesting environmental rather than point-source contamination. Ranitidine multidose vials were kept connected to an automatic compounding machine for up to 48 hours at room temperature after the first dose was drawn, contrary to manufacturer recommendations. Further, preparation of ranitidine infusions was not conducted in accordance with recommendations for risk level 2 sterile i.v. products. The use of contaminated ranitidine multidose vials was the most likely cause of an outbreak of E. cloacae. However, a combination of other factors such as inadequate hand-washing techniques, presence of E. cloacae in the environment, noncompliance with guidelines for the preparation of sterile infusions and medications, and a susceptible population may have contributed to the infections. PMID- 12892029 TI - Impact of emerging technologies on medication errors and adverse drug events. AB - Published evidence on the effects of computerized physician order entry (CPOE), automated dispensing machines (ADMs), bar coding, and computerized medication administration records (CMARs) on medication errors and adverse drug events (ADEs) were reviewed. Emerging technologies have been recommended as potential mechanisms for reducing medication errors. Critical evaluations of the impact of these new technologies on medication errors and other adverse outcomes are lacking. PubMed was searched to identify all peer-reviewed publications linking four technologies (CPOE, ADMs, bar coding, and CMARs) with reductions in medication errors and ADEs and secondary endpoints. All controlled studies that assessed the impact of the technologies were evaluated. The appropriateness of the use of these technologies was also examined. Few studies were identified that evaluated the technologies' impact on these endpoints. Of the evaluated technologies, CPOE was the most studied; however, investigations were limited to selected medical centers. The appropriateness of use of the technologies was evaluated even more infrequently. A literature review revealed a paucity of controlled, generalizable studies confirming the benefits of technologies intended to reduce medication errors and ADEs. Very little evidence on the appropriateness of the use of these technologies was found. PMID- 12892030 TI - Evaluation of a schizophrenia medication algorithm in a state hospital. AB - Provider's practice behaviors before and after physician and staff training in the use of a schizophrenia medication algorithm and the effects of education on physician adherence to the algorithm were evaluated. Medical records of 30 patients admitted between September 1 and November 30, 1999, and 30 patients admitted from September 1 to November 30, 2000, with an admitting and discharge diagnosis of schizophrenia and a minimum length of stay of 14 days were randomly selected and analyzed. Clinical data, including prescribed psychotropic medications and dosages, documentation of target symptoms and severity, adverse drug effects, appropriate clinical ratings, patient's response to treatment, and reason for medication change, were collected and compared with the recommendations in the schizophrenia medication algorithm. Efforts to implement the schizophrenia algorithm included staff education and uniform documentation. Progress notes were evaluated before and after training. After physician and staff training, only 5 of 359 progress notes were written using the recommended documentation form. The number of progress notes containing no documentation of symptoms decreased from 66 to 41, and those documenting three to five target symptoms increased from 74 to 140. Documentation of physician assessment of the presence or absence of adverse effects and their severity decreased from 35.2% to 18.7% and from 22.3% to 17.0%, respectively. Physicians increased the documentation of their clinical global impressions from 12.1% to 20.3%. The recording of medication changes increased twofold, but the difference was not significant. Physician and staff education alone did not significantly alter providers' practice behavior. Inadequate and inconsistent documentation of clinical outcomes made it difficult to assess physician adherence to the treatment algorithm. PMID- 12892031 TI - Unclaimed prescriptions in a clinic with computerized prescriber order entry. PMID- 12892032 TI - Designing a hospital-based pneumococcal vaccination program. PMID- 12892033 TI - Quality assurance in the prescribing of patient-controlled analgesia and long acting opioids. PMID- 12892034 TI - Guidelines for liposomal amphotericin B. PMID- 12892035 TI - Aspirin and cyclooxygenase-2 inhibitors. PMID- 12892036 TI - [Long-term prognosis in patients with single-vessel or double-vessel coronary artery disease: does successful revascularization achieved by coronary angioplasty improve late outcome?]. AB - OBJECTIVES: It is not clear whether revascularization by coronary angioplasty improves long-term prognosis in patients with stable angina, single- or double vessel coronary artery disease. This study investigated the extent of late revascularization achieved by percutaneous transluminal coronary angioplasty(PTCA) and evaluated the long-term effect of revascularization in patients with coronary artery disease. METHODS: This study reviewed 178 patients with single-vessel or double-vessel disease and significant coronary diameter stenosis(American Heart Association classification, 75% or more stenosis), but excluded 35 patients with failed revascularization of the target vessels because of initial failed angioplasty(n = 14) or restenosis(n = 21). The remaining 143 patients were available for assessment. Revascularization was successful at follow-up angiography after the last angioplasty in 105 patients(Group A). The remaining 38 patients received conservative medical therapy(Group B). Kaplan Meier survival curves were used to examine absolute survival difference. RESULTS: One hundred forty-three patients(mean age 64 +/- 9 years) were followed-up for 3.8 +/- 1 years. The patients were 70.6% male, 59.4% had myocardial infarction and 76.9% had single-vessel disease. Anatomical complete revascularization in the chronic phase was accomplished in 80 patients with single-vessel disease(97.6%) and 11 patients with double-vessel disease(47.8%). The event-free survival rate was not significantly different between Group A and Group B(88.6% vs 84.2%, p = NS). Cardiac survival rate decreased significantly in Group B compared to Group A(89.5% vs 99.0%, p < 0.01). This difference in survival rate was due to sudden cardiac death in Group B(7.9% vs 0%, p < 0.05). Non-fatal myocardial infarctions occurred in seven patients(6.7%) with revascularization and in one patient(2.6%) with conservative medical therapy, but with no significant difference. Late successful revascularization improved cardiac survival rate in patients with proximal left anterior descending coronary artery involvement or single-vessel disease. CONCLUSIONS: Cardiac events decreased and long-term prognosis appeared to be a good possibility in patients with single- or double-vessel coronary artery disease if late successful revascularization of target vessel was accomplished. PMID- 12892037 TI - Preventive effect of an antiallergic drug, pemirolast potassium, on restenosis after stent placement: quantitative coronary angiography and intravascular ultrasound studies. AB - OBJECTIVES: The preventive effect of pemirolast against restenosis after coronary stent placement was evaluated. METHODS: Eighty-four patients with 89 de novo lesions who underwent successful coronary stenting were assigned to the pemirolast group(40 patients, 45 lesions) and the control group(44 patients, 44 lesions). Administration of pemirolast(20 mg/day) was initiated from the next morning after stenting and continued for 6 months of follow-up. Quantitative coronary angiography was performed immediately after stenting and at follow-up. Angiographic restenosis was defined as diameter stenosis > or = 50% at follow-up. Intravascular ultrasound study conducted at follow-up angiography was used to measure vessel cross-sectional area(CSA), stent CSA, lumen CSA, neointima CSA(stent CSA--lumen CSA), and percentage neointima CSA(neointima CSA/stent CSA x 100%) at the minimal lumen site. RESULTS: There were no significant differences in baseline characteristics between the two groups. Restenosis rate was significantly lower in the pemirolast group than in the control group(15.0% vs 34.1% of patients, 13.3% vs 34.1% of lesions, p < 0.05, respectively). The intravascular ultrasound study at follow-up(36 lesions in the pemirolast group, 33 in the control group) found no significant differences in vessel CSA and stent CSA between the two groups(17.3 +/- 2.2 vs 16.8 +/- 2.4 mm2, 8.6 +/- 1.9 vs 8.4 +/- 1.7 mm2, respectively). However, lumen CSA was significantly larger in the pemirolast group than in the control group(5.5 +/- 1.3 vs 4.4 +/- 1.1 mm2, p < 0.05). Moreover, neointima CSA and percentage neointima CSA were significantly smaller in the pemirolast group(3.1 +/- 1.1 vs 4.0 +/- 1.2 mm2, p < 0.05 and 36.2 +/- 15.9% vs 47.4 +/- 15.6%, p < 0.01). CONCLUSIONS: Pemirolast has a preventive effect against restenosis after stent placement, possibly by inhibiting neointimal hyperplasia. PMID- 12892038 TI - [Transient retinopathy after acute myocardial infarction treated by reperfusion therapy: clinical background and manifestation]. AB - OBJECTIVES: The severe inflammatory reaction caused by acute myocardial infarction and reperfusion affects both the heart and other remote organs. The occurrence of retinopathy was evaluated in patients with acute myocardial infarction who underwent reperfusion therapy. METHODS: We investigated 29 patients with first acute myocardial infarction who underwent successful reperfusion therapy within 24 hr of the onset. Ophthalmic examinations including visual acuity test and ocular fundoscopy were performed within 3 days, 2 weeks, and then monthly up to 3 months after the onset of acute myocardial infarction. Plasma levels of intercellular adhesion molecule-1(ICAM-1), interleukin-6 and high sensitivity C-reactive protein were measured on admission. RESULTS: Soft exudates around the optic disc appeared in 17(58.6%) of the 29 patients, among whom 5 also developed superficial hemorrhages(17.2%). The retinopathy became most remarkable between 1 to 2 months after the onset of acute myocardial infarction and then faded away without any specific treatment. None of the patients had impairment of visual acuity, although 4 of the 17 patients with retinopathy complained of either blurred vision or metamorphopsia. Hypertension and/or diabetes mellitus tended to be more common in the retinopathy group than in the non-retinopathy group(59% vs 33%, p = 0.096). Plasma ICAM-1 levels were significantly higher than in the non-retinopathy group than in the retinopathy group(p = 0.017). There was no significant difference in plasma levels of interleukin-6 and high sensitivity C-reactive protein between the two groups. CONCLUSIONS: Retinopathy may occur after reperfusion for acute myocardial infarction. The dominant manifestation is transient soft exudates reflecting spotty retinal ischemia, probably due to microvascular obstruction. PMID- 12892039 TI - [Contribution of monocyte chemoattractant protein-1 and c-fms/macrophage colony stimulating factor receptor to coronary artery disease: analysis of human coronary atherectomy specimens]. AB - OBJECTIVES: Primary coronary atherosclerotic lesions(de novo lesions) are a type of inflammatory vascular disease. Restenotic lesions after percutaneous coronary intervention mainly consist of proliferative vascular smooth muscle cells. Recent studies have demonstrated that locally synthesized cytokines, including chemokines, are important in both these coronary lesions. Monocyte chemoattractant protein(MCP)-1 and macrophage colony-stimulating factor(M-CSF) are two of the associated chemokines, but their role in coronary artery disease has not been sufficiently clarified. This study investigated the expression of MCP-1 and c-fms/M-CSF receptor in human coronary tissues. METHODS: Histological and immunohistochemical studies used samples obtained from patients who underwent directional coronary atherectomy(28 de novo lesions and 16 restenotic lesions). The following primary antibodies were used: anti-MCP-1, anti-c-fms, anti macrophages and anti-alpha-smooth muscle actin. RESULTS: Focal accumulation of macrophage-derived foam cells, thrombus, cholesterol clefts and calcification tended to be more frequent in de novo lesions than in restenotic lesions. On the other hand, restenotic lesions mainly consisted of stellate vascular smooth muscle cells and extracellular matrix. The expression of MCP-1-positive cells almost coincided with the macrophages. In contrast, staining for MCP-1 was little seen in the stellate vascular smooth muscle cells. Expression of c-fms was found in both macrophages and stellate vascular smooth muscle cells. Expression patterns of MCP-1 and c-fms exhibited no difference between the two lesion types. CONCLUSIONS: Both MCP-1 and the M-CSF/c-fms system are involved in the atherogenesis of de novo lesions. However, the M-CSF/c-fms system, rather than MCP-1, is more important in the late stage of restenosis. PMID- 12892040 TI - Impedance cardiography and quantitative tissue Doppler echocardiography for evaluating the effect of cardiac resynchronization therapy: a case report. AB - An 83-year-old woman presented with dilated cardiomyopathy. Cardiac resynchronization therapy was performed. Two weeks later, cardiac output and ventricular wall motion were estimated using impedance cardiography and tissue Doppler echocardiography with and without pacing. Cardiac output increased from 3.5 to 4.5 l/min during biventricular pacing with a 120 msec atrioventricular interval. Intraventricular phase difference for contraction decreased from 190 to 150 msec. When the atrioventricular interval was 180 msec, cardiac output and phase difference became 4.6 l/min and 170 msec. These assessments were performed rapidly and non-invasively. New impedance cardiography and tissue Doppler echocardiography are useful to evaluate the effect of cardiac resynchronization therapy. PMID- 12892041 TI - A 64-year-old normotensive woman with right ventricular dilation and hypokinesis. Acute submassive pulmonary thromboembolism with fatal outcome. PMID- 12892042 TI - Evidence that antioxidants prevent the inhibition of Na+,K(+)-ATPase activity induced by octanoic acid in rat cerebral cortex in vitro. AB - The objective of the present study was to investigate the in vitro effects of octanoic acid, which accumulates in medium-chain acyl-CoA dehydrogenase (MCAD) deficiency and in Reye syndrome, on key enzyme activities of energy metabolism in the cerebral cortex of young rats. The activities of the respiratory chain complexes I-IV, creatine kinase, and Na+,K(+)-ATPase were evaluated. Octanoic acid did not alter the electron transport chain and creatine kinase activities, but, in contrast, significantly inhibited Na+,K(+)-ATPase activity both in synaptic plasma membranes and in homogenates prepared from cerebral cortex. Furthermore, decanoic acid, which is also increased in MCAD deficiency, and oleic acid strongly reduced Na+,K(+)-ATPase activity, whereas palmitic acid had no effect. We also examined the effects of incubating glutathione and trolox (alpha tocopherol) alone or with octanoic acid on Na+,K(+)-ATPase activity. Tested compounds did not affect Na+,K(+)-ATPase activity by itself, but prevented the inhibitory effect of octanoic acid. These results suggest that inhibition of Na+,K(+)-ATPase activity by octanoic acid is possibly mediated by oxidation of essential groups of the enzyme. Considering that Na+,K(+)-ATPase is critical for normal brain function, it is feasible that the significant inhibition of this enzyme activity by octanoate and also by decanoate may be related to the neurological dysfunction found in patients affected by MCAD deficiency and Reye syndrome. PMID- 12892043 TI - Numerical and experimental analysis of periodic patterns and sedimentation of lysozyme. AB - This paper deals with experimental investigation, mathematical modelling and numerical simulation of the crystallization processes induced by counter diffusion method of a precipitant agent in a lysozyme protein solution. Novel mathematical strategies are introduced to simulate the experiments and in particular to take into account the kinetics of the growth process and the motion of the crystals due to the combined effect of gravitational force and viscous drag if the sedimenting process is allowed (protein chamber free of gel). Comparison between experimental observations and numerical simulations in the presence of convection and sedimentation and without them provides a validation of the model. The crystal formation in gel results modulated in space. If the gel matrix is not present, convective cells arise in the protein chamber due to local inversions in the density distribution associated to nucleation phenomena. As time passes, these vortex cells migrate towards the top of the protein chamber exhibiting a different wave number according to the distance from the gel interface. The sedimentating particles produce a wake due to depletion of protein from the surrounding liquid. The models and the experiments may represent a useful methodology for the determination of the parameters and conditions that may lead to protein crystallization. PMID- 12892044 TI - Treating the wheezing infant. AB - The management of infants and small children with asthma is a challenging task because of the many issues unique to this age group that deserve special consideration. The diagnosis of asthma is limited by inherent difficulties in obtaining objective measures of lung function and airway inflammation. In persistently symptomatic patients, the decision to initiate controller therapy is not as great an issue as it is in infants and young children with recurrent episodic wheeze in whom early intervention may allow a window of opportunity potentially to alter the course of the disease. The reality is that even if atopy has been consistently implicated in the development of persistent asthma, there is not a well-established set of criteria by which patients who are likely to benefit from early intervention controller therapy can be identified. Hence, large prospective studies need to be performed evaluating the impact of early pharmacologic intervention on the natural history of infantile asthma. Many areas needing investigation involve what medications to use, how best to deliver the medications, and how to monitor the response to treatment. Only a few medications have been approved for use in this population. Long-term studies evaluating available drugs such as inhaled glucocorticoids, LABAs, and the leukotriene modifying agents in young children still need to be performed. PMID- 12892045 TI - [Reversibility of the effects of cadmium on the growth and nitrogen metabolism in the tomato(Lycopersicon esculentum)]. AB - In order to better understand the effects of heavy metals on the growth of plants, we decided to perform recovering experiments by following both chemical and physiological parameters in cadmium pre-stressed tomato seedlings after cadmium had been removed from the nutrient solution. The work shows that cadmium suppression results in resumption of growth activity. The biomass of leaves and stems rose steadily. The increase in root biomass exceeded those of leaves and stems. At the same time, nitrate content was increased to reach the level obtained with unstressed controls. In all the organs studied, the activities of the enzymes involved in the anabolic nitrogen primary assimilation pathways (nitrate reductase (NR), nitrite reductase (NiR) and glutamine synthetase (GS) soared after that cadmium had been removed. While NAD(+)-dependent glutamate dehydrogenase (GDH-NAD+) activity also rose progressively during the recovering time, the cognate NADH-dependent glutamate dehydrogenase (GDH-NADH) activity decreased. This result allows us to propose that the ammonia produced by the stress-induced protein catabolism is detoxified and re-assimilated by the GDH NADH isoenzyme. On the basis of these results, we will discuss the ability of the plant to dilute the effects of pollutants during the recovering period. An important outcome of this work is that a transient contamination of the culture medium by pollutants is not necessarily followed by a significant depreciation in product yield or quality. PMID- 12892046 TI - Characterization of a developmentally regulated oocyst protein from Eimeria tenella. AB - Changes in proteins during sporulation of Eimeria tenella oocysts were investigated. Unsporulated E. tenella oocysts collected from cecal tissue at 7 days postinoculation were sporulated in aerated media at 28 C for 0-48 hr. Gel analysis of soluble protein extracts prepared from oocysts from their respective time points indicated the presence of 2 prominent bands with relative molecular weight (Mr) in the range of 30 kDa and making up 20% of the total protein. These 2 bands, designated as major oocyst proteins (MOPs), were absent or barely detectable by 21 hr of sporulation. MOP bands were weakly reactive with glycoprotein stain but showed no mobility shift on deglycosylation. By gel analysis it was shown that the purified MOPs consisted of 2 bands of Mr 28.7 and 30.1 kDa. However, by matrix-assisted laser deabsorption-time of flight analysis it was shown that masses were about 17% lower. Internal sequence analysis of the 28.7-kDa protein generated 2 peptides of 17 and 14 amino acids in length, consistent with a recently described protein coded by the gam56 gene and expressed in E. maxima gametocytes. Rabbit antibodies made against MOPs were localized to outer portions of sporocysts before excystment and to the apical end of in vitro-derived sporozoites. These same antibodies were found to react with bands of Mr 101 and 65 kDa by Western blot but did not recognize MOPs in soluble or insoluble sporozoite extracts. The data suggest that the MOPs are derived from part of a gametocyte protein similar to that coded by gam56 and are processed during sporulation into sporocyst and sporozoite proteins. Alternatively, the binding of anti-MOP to 101- and 65-kDa proteins may result from alternatively spliced genes as the development of parasite proceeds. PMID- 12892047 TI - How should we select health professionals for studies? AB - The investigation of healthcare professionals' practice patterns has increased sharply, in part driven by the development of practice guidelines; however, the optimal way to select providers is not known. We evaluated three distinct sources of physician specialists for completeness and potential biases. Professional society directories, which are frequently used to identify providers, provided biased populations. A national registry, the American Medical Association master file, produced the most comprehensive, least-biased single source. PMID- 12892048 TI - Vitamin E treatment for tardive dyskinesia. Veterans Affairs Cooperative Study #394 Study Group. AB - BACKGROUND: Several short-term, controlled trials have documented the efficacy of vitamin E in treating tardive dyskinesia. However, the persistent nature of the disease prompted us to perform a multicenter, longer-term trial of vitamin E. METHODS: The study was a prospective, randomized, 9-site trial of up to 2 years of treatment with d-vitamin E (1600 IU/d) vs matching placebo. One hundred fifty eight subjects with tardive dyskinesia who were receiving neuroleptic medications were enrolled. The blinded assessments performed were clinical (Abnormal Involuntary Movements Scale, Barnes Akathisia Scale, and Modified Simpson-Angus [for Extrapyramidal Symptoms] Scale) and electromechanical assessments of movement disorders, psychiatric status (Brief Psychiatric Rating Scale), and functioning (Global Assessment of Functioning). There were no significant differences in baseline demographic characteristics or in study assessments between the group that received vitamin E and the group that received placebo. RESULTS: Vitamin E was well tolerated and subject compliance with medication was good and similar between treatment groups. One hundred seven subjects (70% of those receiving vitamin E and 66% of subjects receiving placebo) completed at least 1 year of treatment. There were no significant effects of vitamin E on total scores or subscale scores for the AIMS, electromechanical measures of dyskinesia, or scores from the other 4 scales. CONCLUSION: This long-term, randomized trial of vitamin E vs placebo found no evidence for efficacy of vitamin E in the treatment of tardive dyskinesia. PMID- 12892049 TI - FIZZ1 and Ym as tools to discriminate between differentially activated macrophages. AB - Although it is well-established that macrophages can occur in distinct activation states, the molecular characteristics of differentially activated macrophages, and particularly those of alternatively activated macrophages (aaMphi), are still poorly unraveled. Recently, we demonstrated that the expression of FIZZ1 and Ym is induced in aaMphi as compared with classically activated macrophages (caMphi), elicited in vitro or developed in vivo during infection with Trypanosoma brucei brucei. In the present study, we analyzed the expression of FIZZ1 and Ym in caMphi and aaMphi elicited during Trypanosoma congolense infection and show that the use of FIZZ1 and Ym for the identification of aaMphi is not limited to T. b. brucei infection and is independent of the organ sources from which macrophages are obtained. We also demonstrate that FIZZ1 can be used to discriminate between different populations of aaMphi. Furthermore, we studied the effects of various stimuli, and combinations thereof, on the expression of FIZZ1 and Ym in macrophages from different mouse strains and demonstrate that regulation of the expression of FIZZ1 and Ym in macrophages is not dependent on the mouse strain. Finally, we show that these genes can be used to monitor the macrophage activation status without the need to obtain pure macrophage populations. PMID- 12892050 TI - [Dichloroacetate treatment for adult patients with mitochondrial disease]. AB - We report beneficial and adverse effects of sodium dichloroacetate (DCA) in three adult Japanese patients with mitochondrial disease: a 21-year-old male with involuntary movements, optic atrophy, hearing loss, and convulsions (patient 1), a 28-year-old man with mental deterioration, hemianopia, hearing disturbance, and convulsions (patient 2), and a 50-year-old woman with hearing disturbance, generalized muscle atrophy, and insulin dependent diabetes mellitus (patient 3). A3243G mutation was found in patient 2 and patient 3. Oral administration of DCA improved consciousness level and gait disturbances in patient 1, and ameliorated headaches, easy fatiguability, and muscle cramps in patient 2 and patient 3. DCA normalized high levels of lactate and pyruvate in blood and cerebrospinal fluids in all three patients. In patient 3, daily insulin needs decreased from 38 to 24 units, and urine C peptide increased from an undetectable level to 16 micrograms/day. In patient 1, DCA 23 mg/kg/day had been beneficial without adverse effects and he became free of convulsions for more than 32 months. However, despite of normal lactate and pyruvate, unsteady gait and lethargy developed after 50 mg/kg/day treatment for two months and one month in patient 2 and patient 3, respectively. In both patients, deep tendon reflexes disappeared and Romberg sign became positive. Nerve conduction studies confirmed sensory dominant polyneuropathy and electroencephalogram showed diffuse slow basic activities. Cessation of DCA resulted in recovery of gait and consciousness, but sensory nerve action potentials did not recover in one month. Long term treatment of 50 mg/kg/day DCA may affect adversely the peripheral and central nervous systems in adult patients. Although effective plasma DCA concentration was previously reported as 25-160 micrograms/ml in patients under 18 years old, plasma DCA concentration of 10.2 micrograms/ml was sufficient in patient 1. We recommend lower dose of DCA in adult patients than in child patients. PMID- 12892051 TI - Epidemiology of sporadic bloody diarrhea in rural Western Kenya. AB - We conducted laboratory-based surveillance and a case-control study to characterize the epidemiology of bloody diarrhea in rural Western Kenya. From May 1997 through April 2001, we collected stool from 451 persons with bloody diarrhea presenting to four rural clinics. Cultures of 231 (51%) specimens yielded 247 bacterial pathogens: 198 Shigella (97 S. flexneri, 41 S. dysenteriae type 1, 39 S. dysenteriae type non-1, 13 S. boydii, 8 S. sonnei), 33 Campylobacter, 15 non typhoidal Salmonella, and 1 Vibrio cholerae O1. More than 90% of the isolates (excluding Campylobacter) were resistant to trimethoprim-sulfamethoxazole and tetracycline, and more than 80% were resistant to ampicillin. Most (74%) ill persons received medication to which their isolate was resistant. Drinking Lake Victoria water and sharing latrines between multiple households increased risk of bloody diarrhea. Washing hands after defecating was protective. Providing safe drinking water and more latrines, and promoting hand washing could reduce the burden of illness from bloody diarrhea while limiting injudicious antimicrobial use. PMID- 12892053 TI - Severe acute respiratory syndrome (SARS) virus. PMID- 12892052 TI - cAMP-mediated suppression of a Th1 clone associated with an alteration of the intracellular redox environment. AB - We have shown previously that the elevation of intracellular cAMP in antigen or anti-CD3-activated murine Th1 clones in the absence of antigen inhibits antigen induced proliferation and the production of IL-2 by H2O2-mediated oxidation of p56lck and inhibits antigen-induced production of interferon-gamma by the induction of intracellular nitric oxide. Moreover, activated Th1 clones are resistant to cAMP-induced suppression. These results suggest that the immunosuppression of Th1 cells mediated by elevated intracellular cAMP is associated with an alteration in the intracellular oxidation/reduction environment. Here we report that the culture of an antigen or anti-CD3-activated murine Th1 clone with the adenylcyclase agonist forskolin (FSK) in the absence of antigen reduces the activity of intracellular catalase, and diminishes levels of intracellular reduced glutathione (GSH). Resting cells resistant to cAMP-induced suppression have higher intracellular GSH levels than antigen-activated cells susceptible to cAMP-induced suppression. The results provide further evidence that cAMP-induced suppression of Th1 clones is mediated by profound alterations in the intracellular redox environment and may be used to selectively inactivate Th1 cells activated by antigen. PMID- 12892054 TI - Congress "fixed the mistake"--now, they must "fix the formula.". PMID- 12892055 TI - Isolations of yellow fever virus from Haemagogus leucocelaenus in Rio Grande do Sul State, Brazil. AB - Following howling monkey (Alouatta caraya) deaths and yellow fever (YF) antigen detection by immunohistochemistry in the liver sample of a dead monkey in April and May 2001 in the municipalities of Garruchos and Santo Antonio das Missoes, Rio Grande do Sul State, Brazil, epidemiological field investigations were initiated. Two strains of YF virus were isolated in suckling mice from 23 Haemagogus (Conopostegus) leucocelaenus Dyar & Shannon mosquitoes collected from the study sites. The YF virus was isolated from this species in the 1930s in Brazil and in the 1940s in Colombia. No human cases were reported during the current epizootic outbreak. The YF virus isolation and the absence of Hg. (Haemagogus) janthinomys Dyar from the area suggest that Hg. leucocelaenus may be a secondary YF vector and play an important role in the epidemiology of this disease in the Southern Cone. PMID- 12892056 TI - [A review of the study on swainsonine--toxic elements of locoweed]. PMID- 12892057 TI - [Is radiotherapy indicated for ductal carcinoma of the breast?]. PMID- 12892058 TI - [Terahertz rays: at the threshold for clinical practice]. PMID- 12892059 TI - [Fowl plague: a potential danger also for humans]. PMID- 12892060 TI - Spreading of HIV-1 subtype G and envB/gagG recombinant strains among injecting drug users in Lisbon, Portugal. AB - We have evaluated the genetic diversity of HIV-1 strains infecting injecting drug users (IDUs) in Lisbon, Portugal. Heteroduplex mobility assay and/or phylogenetic analysis revealed that env (C2V3C3 or gp41) subtype B is present in 63.7% of the 135 viral samples studied, followed by subtypes G (23.7%), A (6.7%), F (5.2%), and D (0.7%). Similar analysis of gag (p24/p7) performed on 91 of the specimens demonstrated that 49.5% of the infections were caused by subtype G viruses; other gag subtypes identified were B (39.5%), F (3.3%), A and D (1.1.% each), and the recombinant circulating form CRF02_AG (5.5%). Discordant env/gag sub-types were detected in 34.1% of the strains and may reflect the presence of dual infections and/or recombinant viruses. The presumptive B/G recombinant form was highly predominant (21 of 31). The genetic pattern of HIV-1 subtype B and G strains is suggestive of multiple introductions and recombination episodes and of a longstanding presence of both subtypes in the country. C2V3C3 amino acid sequences from IDU-derived subtype G viruses presented highly significant signatures, which distinguish the variants from this transmission group. The unusually high prevalence of subtype G sequences (34.1%), independent of the geographic origin of the infected individuals, makes this IDU HIV-1 epidemic unique. PMID- 12892061 TI - Brazilian HTLV type 2a strains from intravenous drug users (IDUs) appear to have originated from two sources: Brazilian Amerindians and European/North American IDUs. AB - In Brazil, HTLV-2 has been detected in blood donors, in intravenous drug users (IDUs) from urban areas, and in Amerindians living in the Amazon basin. Of the three main HTLV-2 subtypes (2a, 2b, and 2d) only subtype 2a has been detected in Brazil. However, a molecular variant of subtype 2a (also called HTLV-2c) characterized by an extended Tax protein has been isolated from Brazilian blood donors, IDUs, and Indians. Here, we analyzed HTLV-2 isolates from 10 IDUs and a Chilean woman living in Salvador, Bahia, Brazil. Sequencing of env, pX, and long terminal repeat (LTR) genes demonstrated that 10 of the isolates are related to the Brazilian subtype 2a molecular variant described previously. We show that most HTLV-2a Brazilian strains comprise a phylogenetic group harboring a considerable degree of diversity within the env region but not within the LTR region. Interestingly, we demonstrated for the first time in Brazil the presence of a subtype 2a in IDUs that is closely related to the prototype Mo but distinct from the Brazilian 2a molecular variant. PMID- 12892062 TI - Natural polymorphisms of protease in protease inhibitor-naive HIV-1 infected patients in Korea: a novel L63M in subtype B. AB - To establish a baseline for monitoring resistance mutation to protease inhibitors (PI), we determined protease(PR) sequences in peripheral blood mononuclear cells obtained from 43 PI-naive Korean HIV-1 infected patients. Interestingly, phylogenetic analysis revealed that 41 of the sequences belonged to subtype B, one belonged to subtype A, and one was unique, not clustering with any subtype. Thirty-one (76%) of the 41 sub-type B sequences formed a subclade within subtype B, a so-called "Korean B cluster." Polymorphisms were observed at 34 (34.3%) of the PR codons. One patient (2.3%) harbored a primary resistance-conferring mutation, L90M along with L63P and A71V, and all 43 strains showed some secondary associated with drug resistance. The percentage of patients with 7, 5, 4, 3, 2, and 1, resistance mutations were 2%, 2%, 14%, 23%,37%, and 9%, respectively. A novel polymorphism in subtype B, L63M was detected in two patients. Another patient showed a gross deletion (257 bp) after codon 91. The average genetic distance of the 41 subtype B sequences to the HXB2 sequence was 3.0% (range, 1.0 5.1%). Six hemophiliacs were infected with a domestic strain of HIV-1 subtype B, while the other two hemophiliacs were infected with nondomestic subtype B and had lived outside Korea. Although this is the first report on the molecular nature of PR in Korea, there is also a need to establish baselines for nonsubtype B HIV-1. PMID- 12892063 TI - On arteriogenesis--a reply. PMID- 12892064 TI - Syndromic surveillance: a local perspective. PMID- 12892065 TI - [Accident videos lower risk of injury and promote proper behavior]. PMID- 12892066 TI - [Nordic walking. Also suitable in rehabilitation]. PMID- 12892067 TI - [Athletic injuries in middle-distance and long-distance runners]. PMID- 12892068 TI - Orthopaedic proceedings. 2001, 2002. PMID- 12892069 TI - ISEH 2003. Abstracts of the 32nd Annual Meeting of the International Society for Experimental Hematology. Paris, France, July 5-8, 2003. PMID- 12892070 TI - Neoepitopes as biomarkers of cartilage catabolism. AB - Progressive degradation of articular cartilage is a central feature of arthritis and a major determinant of long term joint dysfunction. There are no treatments able to halt the progression of cartilage destruction presently available, and monitoring the benefit of potential therapies is hampered by our inability to measure the "health" of articular cartilage. Serial radiographic assessment of joint space narrowing, the current gold standard, requires measurements over a prolonged time (1-5 years) and is prone to technical difficulties. Other strategies for evaluating cartilage degradation are needed to enable both short and long term monitoring of disease progression and response to therapy. One avenue that holds promise is the use of biomarkers that accurately reflect the degradative state of the articular cartilage. Antibodies that recognise terminal amino acid sequences generated by proteolysis at specific sites in the core protein of both aggrecan and type II collagen (neoepitope antibodies) have become available in recent years. These antibodies have been invaluable for identifying the proteinases responsible for cartilage breakdown both in vitro and in vivo. The presence of neoepitope sequences generated by specific metalloenzyme cleavage of aggrecan and type II collagen correlates well with the progression of cartilage degeneration, both in vitro and in mouse models of arthritis. Preliminary results with quantitative assays of type II collagen neoepitopes suggest that they may be useful markers of joint disease in humans. Long term studies correlating neoepitope concentration with clinical and radiographic disease are now required to validate the utility of neoepitopes as surrogate markers of cartilage degeneration and joint disease. PMID- 12892071 TI - Best evidence in anesthetic practice. Goal-directed therapy with the pulmonary artery catheter is not better than standard therapy. PMID- 12892073 TI - Abstracts of the Society for Technology in Anesthesia 2003 Meeting, the 3rd Annual International Meeting on Medical Simulation (2003), and Computers in Anesthesia XXIII (2002). PMID- 12892072 TI - The Tg.AC mouse model passes test by failing to respond. PMID- 12892074 TI - Summary of notifiable diseases--United States, 2001. AB - The MMWR Summary of Notifiable Diseases, United States, 2001 contains, in tabular and graphic form, the official statistics for the reported occurrence of nationally notifiable diseases in the United States for 2001. These statistics are collected and compiled from reports sent by state health departments to the National Notifiable Diseases Surveillance System (NNDSS), which is operated by CDC in collaboration with the Council of State and Territorial Epidemiologists (CSTE). PMID- 12892076 TI - Abstracts of the XXIst International Conference on Yeast Genetics and Molecular Biology. Gothenburg, Sweden, 7-12 July 2003. PMID- 12892075 TI - [Professional health in China's township enterprises: development and challenge]. PMID- 12892077 TI - [Heat stress protein: progress and prospects]. PMID- 12892079 TI - Ten-year cumulative index, 1993-2002. PMID- 12892078 TI - Summaries of notifiable diseases in the United States, 2001. PMID- 12892080 TI - Graphs and maps for selected notifiable diseases in the United States. PMID- 12892081 TI - Historical summaries of notifiable diseases in the United States, 1970-2001. PMID- 12892083 TI - The Women's Health Data Book. A profile of women's health in the United States. PMID- 12892082 TI - Abstracts of the 11th British Academic Conference in Otolaryngology. Birmingham, United Kingdom, 2-5 July 2003. PMID- 12892084 TI - No ischemic preconditioning in heterozygous connexin 43-deficient mice--a further in vivo study. PMID- 12892086 TI - Coping with sudden incidents of public health, new ordinances were issued. PMID- 12892085 TI - RCM evidence to the Pay Review Body. PMID- 12892087 TI - Scotland's think-tank day. PMID- 12892088 TI - [Environment and health: towards integration. Views from environmental agencies]. PMID- 12892089 TI - Abstracts of the 4th International Conference on Homocysteine Metabolism. Basel, Switzerland, June 29-July 3, 2003. PMID- 12892091 TI - Notifiable diseases annual summary, 2002. PMID- 12892090 TI - SARS treatment: experience from a team in Guangdong, China. PMID- 12892092 TI - Measures against SARS took effect in China. PMID- 12892093 TI - Who's who in security. PMID- 12892094 TI - Caffeine intake and the risk of spontaneous abortion. PMID- 12892095 TI - [Planned immunology in China: establishment and improvement]. PMID- 12892096 TI - Congenital and acquired long QT syndromes. PMID- 12892097 TI - Chalk bones and pathological fractures: case report and review of the literature. PMID- 12892098 TI - [Injury: a problem threatening the health of young generation]. PMID- 12892099 TI - Intellectual impairment and blood lead levels. PMID- 12892100 TI - Intellectual impairment and blood lead levels. PMID- 12892101 TI - Obesity and cancer. PMID- 12892102 TI - Obesity and cancer. PMID- 12892103 TI - Obesity and cancer. PMID- 12892104 TI - Obesity and cancer. PMID- 12892106 TI - [The recent state of study on salivary gland diseases in China]. PMID- 12892105 TI - Fibroblast growth factor 23 in oncogenic osteomalacia and X-linked hypophosphatemia. PMID- 12892107 TI - Adenosine-induced ventricular flutter? PMID- 12892108 TI - Validation of publication of new names and new combinations previously effectively published outside the IJSEM. PMID- 12892109 TI - Notification that new names and new combinations have appeared in volume 53, part 2, of the IJSEM. PMID- 12892110 TI - Phylogenetic taxonomy of the family Chlorobiaceae on the basis of 16S rRNA and fmo (Fenna-Matthews-Olson protein) gene sequences. AB - A new taxonomy of the green sulfur bacteria is proposed, based on phylogenetic relationships determined using the sequences of the independent 16S rRNA and fmo (Fenna-Matthews-Olson protein) genes, and supported by the DNA G + C content and sequence signatures. Comparison of the traditional classification system for these bacteria with their phylogenetic relationship yielded a confusing picture, because properties used for classification (such as cell morphology, photosynthetic pigments and substrate utilization) do not concur with their phylogeny. Using the genetic information available, strains and species assigned to the genera Chlorobium, Pelodictyon and Prosthecochloris are considered, and the following changes are proposed. Pelodictyon luteolum is transferred to the genus Chlorobium as Chlorobium luteolum comb. nov. Pelodictyon clathratiforme and Pelodictyon phaeoclathratiforme are transferred to the genus Chlorobium and combined into one species, Chlorobium clathratiforme comb. nov. The name Pelodictyon will become a synonym of Chlorobium. Strains known as Chlorobium limicola subsp. thiosulfatophilum that have a low DNA G + C content (52-52.5 mol%) are treated as strains of Chlorobium limicola; those with a high DNA G + C content (58.1 mol%) are transferred to Chlorobaculum gen. nov., as Chlorobaculum thiosulfatiphilum sp. nov. Chlorobium tepidum is transferred to Chlorobaculum tepidum comb. nov., and defined as the type species of the genus Chlorobaculum. Strains assigned to Chlorobium phaeobacteroides, but phylogenetically distant from the type strain of this species, are assigned to Chlorobium limicola and to Chlorobaculum limnaeum sp. nov. Strains known as Chlorobium vibrioforme subsp. thiosulfatophilum are transferred to Chlorobaculum parvum sp. nov. Chlorobium chlorovibrioides is transferred to 'Chlorobaculum chlorovibrioides' comb. nov. The type strain of Chlorobium vibrioforme is phylogenetically related to Prosthecochloris, and is therefore transferred to this genus as Prosthecochloris vibrioformis comb. nov. Consequently, the name Chlorobium vibrioforme will become a synonym of Prosthecochloris vibrioformis, and other strains that were assigned to this species are now considered to belong to Chlorobium luteolum, Chlorobium phaeovibrioides and 'Chlorobaculum chlorovibrioides', according to their phylogenetic relatedness. PMID- 12892111 TI - Seven novel species of Acinetobacter isolated from activated sludge. AB - Thirteen isolates of Acinetobacter were obtained from activated sludge plants in Victoria, Australia. Earlier 16S-23S rDNA genomic fingerprinting and partial 16S rDNA sequence data had suggested that these isolates might contain previously undescribed species. This view was confirmed here. A polyphasic taxonomic approach involving phenotypic characterization, near-complete 16S rDNA sequence data and DNA-DNA hybridization analyses support the view that seven novel genomic species can be differentiated in this group of isolates. However, when fluorescence in situ hybridization (FISH) studies were performed with a 16S-rRNA targeted probe specific for the genus Acinetobacter, used to identify Acinetobacter in activated sludge plants, all these strains responded positively. This suggests that these isolates would not have been missed in earlier FISH studies where their role as polyphosphate-accumulating bacteria has been questioned. This report describes these isolates and proposes that they be named Acinetobacter baylyi (type strain B2T = DSM 14961T = CIP 107474T), Acinetobacter bouvetii (type strain 4B02T = DSM 14964T = CIP 107468T), Acinetobacter grimontii (type strain 17A04T = DSM 14968T = CIP 107470T), Acinetobacter tjernbergiae (type strain 7N16T = DSM 14971T = CIP 107465T), Acinetobacter towneri (type strain AB1110T = DSM 14962T = CIP 107472T), Acinetobacter tandoii (type strain 4N13T = DSM 14670T = CIP 107469T) and Acinetobacter gerneri (type strain 9A01T = DSM 14967T = CIP 107464T). PMID- 12892112 TI - Eubacterium pyruvativorans sp. nov., a novel non-saccharolytic anaerobe from the rumen that ferments pyruvate and amino acids, forms caproate and utilizes acetate and propionate. AB - Two similar gram-positive rods were isolated from 10(-6) dilutions of ruminal fluid from a sheep receiving a mixed grass hay/concentrate diet, using a medium containing pancreatic casein hydrolysate as sole source of carbon and energy. The isolates did not ferment sugars, but grew on pyruvate or trypticase, forming caproate as the main fermentation product and valerate to a lesser extent. Acetate and propionate were utilized. One of these strains, I-6T, was selected for further study. Strain I-6T was a non-motile coccal rod, 1.2 x 0.4 microm, with a gram-positive cell wall ultrastructure and a G + C content of 56.8 mol%. No spores were visible, and strain I-6T did not survive heating at 80 degrees C for 10 min. Its rate of NH3 production was 375 nmol (mg protein)(-1) min(-1), placing it in the 'ammonia-hyperproducing' (or HAP) group of ruminal bacteria. 16S rDNA sequence analysis (1296 bases) indicated that it represents a novel species within the 'low-G + C' gram-positive group, for which the name Eubacterium pyruvativorans sp. nov. is proposed. Among cultivated bacteria, strain I-6T was most closely related (89% identity) to other asaccharolytic Eubacterium isolates from the mouth and the rumen. It was 98% identical to uncultured bacterial sequences amplified by others from ruminal digesta. PMID- 12892113 TI - Alkaliphilus crotonatoxidans sp. nov., a strictly anaerobic, crotonate dismutating bacterium isolated from a methanogenic environment. AB - Two bacterial strains were isolated from methanogenic butyrate-oxidizing mixed cultures. The cells were straight to slightly curved, gram-positive rods that were motile by means of multiple flagella and formed endospores. Growth was observed in the temperature range 15-45 degrees C (optimum 37 degrees C) and pH range 5.5-9.0 (optimum pH 7.5). The novel isolates were strictly anaerobic chemo organotrophs capable of utilizing yeast extract, peptone, tryptone and a variety of sugars and organic acids, but not glucose. None of the accessory electron acceptors tested (elemental sulfur, thiosulfate or fumarate) improved growth, except crotonate, which was dismutated to butyrate and acetate. The G + C content of the DNA of one of the isolates, strain B11-2T, was 30.6 mol%. Phylogenetic analysis based on 16S rDNA sequence similarity between strain B11-2T and some other strictly anaerobic, spore-forming bacteria indicated that the novel isolates represented a species in cluster XI within the low-GC gram-positive bacteria, being most closely related to Alkaliphilus transvaalensis JCM 10712T. DNA-DNA relatedness between strain B11-2T and A. transvaalensis JCM 10712T was 21%. On the basis of physiological and molecular properties, and cellular fatty acid and cell wall compositions, the novel isolates are proposed to represent a novel species of the genus Alkaliphilus, for which the name Alkaliphilus crotonatoxidans is proposed (type strain B11-2T=AS 1.2897T=JCM 11672T). PMID- 12892114 TI - Leifsonia rubra sp. nov. and Leifsonia aurea sp. nov., psychrophiles from a pond in Antarctica. AB - Two unique psychrophilic strains (CMS 76rT and CMS 81yT) were isolated from a cyanobacterial mat sample from a pond in Wright Valley, McMurdo, Antarctica. Both isolates were assigned to the genus Leifsonia, since they were gram-positive, curved rods, non-motile, catalase-positive, contained DL-2,4-diaminobutyric acid, menaquinone MK-11, phosphatidylglycerol and diphosphatidylglycerol, had a high content of anteiso- and iso-branched fatty acids and had a DNA G + C content of 64-66 mol%. In addition, both isolates were related to the five reported species of Leifsonia at a level of about 95-96% 16S rDNA sequence similarity and differed from one another by 2.5%. Strains CMS 76rT and CMS 81yT also differed from one another in many other phenotypic characteristics and exhibited only 30% relatedness at the DNA-DNA level, thus indicating that they represent two different species. Furthermore, these two isolates also showed many distinct differences with respect to the reported species of Leifsonia in terms of their phenotypic characteristics, biochemical properties, chemotaxonomic features, sensitivity to various antibiotics and 16S rDNA similarity, clearly indicating that strains CMS 76rT (= MTCC 4210T = DSM 15304T = CIP 107783T) and CMS 81yT (= MTCC 4657T = DSM 15303T = CIP 107785T) represent the type strains of two novel species of Leifsonia, for which the names Leifsonia rubra sp. nov. and Leifsonia aurea sp. nov. are proposed. PMID- 12892115 TI - Rhodoglobus vestalii gen. nov., sp. nov., a novel psychrophilic organism isolated from an Antarctic Dry Valley lake. AB - A novel, psychrophilic, gram-positive bacterium (designated strain LV3T) from a lake near the McMurdo Ice Shelf, Antarctica, has been isolated and characterized. This organism formed red-pigmented colonies, had an optimal growth temperature of 18 degrees C and grew on a variety of media between -2 and 21 degrees C. Scanning electron micrographs of strain LV3T that showed small rods with unusual bulbous protuberances during all phases of growth were of particular interest. The G + C content of the genomic DNA was approximately 62 mol%. The cell walls contained ornithine as the diamino acid. The major fatty acids were anteiso-C15:0, iso C16:0 and anteiso-C17:0. Cells grown at -2 degrees C contained significant amounts of anteiso-C15:1. The major menaquinones found in strain LV3T were MK-11 and MK-12. Phylogenetic analysis of the 16S rRNA gene sequence indicated that strain LV3T was a member of the family Microbacteriaceae and related to, but distinct from, organisms belonging to the genera Agreia, Leifsonia and Subtercola. In addition, alignments of 16S rRNA sequences showed that the sequence of strain LV3T contained a 13 bp insertion that was found in only a few related sequences. Based on the low growth temperature, unusual cell shape, distinct 16S rRNA gene sequence and structure and cell-wall amino acid and menaquinone compositions, Rhodoglobus vestalii gen. nov., sp. nov. is proposed, with the type strain LV3T (= ATCC BAA-534T = CIP 107482T). PMID- 12892116 TI - Reclassification of ATCC 9341 from Micrococcus luteus to Kocuria rhizophila. AB - Strain ATCC 9341, currently known as Micrococcus luteus, has been designated as a quality-control strain in a number of applications. It is also cited as the standard culture in several official methods and manuals, as well as the Code of Federal Regulations. Over the years, it has become apparent that ATCC 9341 does not resemble other M. luteus strains; however, its phenotypic characteristics alone were ambiguous. Recently, a polyphasic study was performed in which molecular data were combined with cytochemical properties and physiological characteristics. The results clearly indicate that ATCC 9341 is a member of the genus Kocuria. Thus, it is proposed to reclassify ATCC 9341 as Kocuria rhizophila and to alert users worldwide of this name change. PMID- 12892117 TI - Nocardia caishijiensis sp. nov., a novel soil actinomycete. AB - A soil isolate, strain F829T, which had provisionally been assigned to the genus Nocardia, was subjected to a polyphasic taxonomic study. An almost complete 16S rDNA sequence was determined for this strain; the sequence was aligned with available sequences for nocardiae and phylogenetic trees were inferred using three tree-making algorithms. The organism showed a combination of phenotypic properties typical of nocardiae and formed a distinct phyletic line within the evolutionary radiation of species of the genus Nocardia, being most closely related to Nocardia asteroides ATCC 19247T. Strain F829T was readily distinguished from representatives of species of Nocardia with validly published names on the basis of phenotypic data, notably from the type strain of N. asteroides. It is proposed that the organism be recognized as a novel species of Nocardia, Nocardia caishijiensis sp. nov. The type strain is F829T (=AS 4.1728T=JCM 11508T). PMID- 12892118 TI - Transfer of Hongia koreensis Lee et al. 2000 to the genus Kribbella Park et al. 1999 as Kribbella koreensis comb. nov. AB - The taxonomic status of Hongia koreensis was investigated by molecular systematic methods. On the basis of 16S rDNA phylogeny, H. koreensis was closely associated with Kribbella flavida and Kribbella sandramycini and formed a monophyletic clade with these species. These three taxa shared over 98% 16S rDNA sequence similarity and many chemotaxonomic properties, which strongly indicates that they belong to the same genus. DNA-DNA pairing was employed to elucidate the genomic relatedness among these taxa. H. koreensis represented a distinct genomic species that can be differentiated from members of the genus Kribbella. Physiological characteristics and phospholipid and cellular fatty acid compositions can be also used to separate H. koreensis from species of the genus Kribbella. On the basis of data presented in this and earlier studies, it is proposed that H. koreensis Lee et al. 2000 should be transferred to the genus Kribbella Park et al. 1999 as Kribbella koreensis comb. nov. PMID- 12892119 TI - Corynebacterium sphenisci sp. nov., isolated from wild penguins. AB - Six unidentified gram-positive, rod-shaped organisms recovered from the cloacae of apparently healthy wild penguins were characterized by phenotypic and molecular taxonomic methods. Chemotaxonomic investigations revealed the presence of a cell wall based on meso-diaminopimelic acid and long-chain cellular fatty acids of the straight-chain saturated and monounsaturated types, consistent with the genus Corynebacterium. Corynomycolic acids, which are characteristic of the genus, were also detected, albeit in small amounts. Comparative 16S rRNA gene sequencing studies showed that the unidentified organisms were phylogenetically related to corynebacteria and represent a novel subline associated with a small subcluster of species that includes Corynebacterium xerosis, Corynebacterium amycolatum and Corynebacterium freneyi. The unknown isolates were readily distinguished from their closest phylogenetic relatives and all other Corynebacterium species with validly published names by using a combination of biochemical and chemotaxonomic criteria. Based on both phenotypic and 16S rRNA gene sequence considerations, it is proposed that the unknown isolates recovered from penguins be classified as a novel species in the genus Corynebacterium, Corynebacterium sphenisci sp. nov. The type strain is CECT 5990T (= CCUG 46398T). PMID- 12892120 TI - Zooshikella ganghwensis gen. nov., sp. nov., isolated from tidal flat sediments. AB - Two red pigment-producing bacterial strains with a metallic green sheen were isolated from a sediment sample of getbol, the Korean tidal flat. Phylogenetic analysis based on 16S rDNA sequences showed that these isolates represent a phyletic lineage within the gamma-Proteobacteria that is distantly related to the genus Hahella. No bacterial species with validly published names showed > or = 92% 16S rRNA similarity with the getbol isolates. The strains were gram-negative, chemo-organotrophic, aerobic and required NaCl (1-7%) for growth. They produced pigments with maximum absorption at 540 nm, which indicated the presence of prodigiosin, a well-known red pigment previously detected in Serratia marcescens. The major isoprenoid quinone was ubiquinone-9. The predominant cellular fatty acids were saturated and monounsaturated straight-chain fatty acids. The DNA G + C contents ranged from 40 to 42 mol%. The combination of physiological, biochemical and chemotaxonomic data clearly separated the test strains from other phylogenetically related genera in the gamma-Proteobacteria. On the basis of polyphasic evidence from this study, it is proposed that the two getbol isolates should be classified in a novel genus, Zooshikella gen. nov., as Zooshikella ganghwensis sp. nov. PMID- 12892121 TI - Characterization of novel psychrophilic clostridia from an Antarctic microbial mat: description of Clostridium frigoris sp. nov., Clostridium lacusfryxellense sp. nov., Clostridium bowmanii sp. nov. and Clostridium psychrophilum sp. nov. and reclassification of Clostridium laramiense as Clostridium estertheticum subsp. laramiense subsp. nov. AB - Taxonomic studies were performed on four strains (D-1/D-an/IIT, C/C-an/B1T, A-1/C an/C1T and A-1/C-an/IT) of anaerobic, gram-positive, spore-forming bacteria originally isolated from a mat sample retrieved from a shallow, moated area around Lake Fryxell, an Antarctic freshwater lake. Phylogenetic analyses based on 16S rRNA gene sequence data indicated that these strains are affiliated with cluster I clostridia and form a coherent group with Clostridium estertheticum and Clostridium laramiense. Similarity values among 16S rRNA gene sequences within this assemblage ranged between 96.7 and 99.8%. Despite the close phylogenetic relationship, several distinguishing phenotypic traits were found among the novel strains using a polyphasic approach. All strains were psychrophilic, but the temperature optimum for growth differed markedly, ranging from 4 to 16 degrees C. In addition, substrate utilization patterns, fermentation end products, cellular fatty acid profiles and morphological traits enabled a clear differentiation between the strains. DNA-DNA hybridization experiments revealed that each of the four novel strains represents a distinct species, with DNA-DNA similarity values to related strains in the range 16-62%. In contrast, the type strains of C. estertheticum and C. laramiense shared 79% DNA-DNA similarity, indicating a close relationship at the species level. On the basis of genetic and phenotypic properties, it is proposed to designate four novel species of the genus Clostridium to harbour the newly isolated strains: Clostridium frigoris sp. nov. (type strain D-1/D-an/IIT=DSM 14204T=ATCC BMAA-579T), Clostridium lacusfryxellense sp. nov. (type strain C/C-an/B1T=DSM 14205T=ATCC BAA-580T), Clostridium bowmaniisp. nov. (type strain A-1/C-an/C1T=DSM 14206T=ATCC BAA-581T) and Clostridium psychrophilum sp. nov. (type strain A-1/C-an/IT=DSM 14207T=ATCC BAA-582T). It is also proposed to unite C. laramiense and C. estertheticum under C. estertheticum. The subspecies C. estertheticum subsp. laramiense subsp. nov. is established, represented by strain ATCC 51254T (=DSM 14884T). The type strain of C. estertheticum subsp. estertheticum remains NCIMB 12511T (=DSM 8809T). PMID- 12892122 TI - Parvularcula bermudensis gen. nov., sp. nov., a marine bacterium that forms a deep branch in the alpha-Proteobacteria. AB - Two bacterial strains, HTCC2503T and HTCC2517, were isolated from the Bermuda Atlantic Time Series Station in the western Sargasso Sea, Atlantic Ocean, by new high-throughput culture methods that rely on dilution to extinction in very-low nutrient media. Characterization of the two strains by polyphasic approaches revealed that they belonged to the same species. These isolates are gram negative, strictly aerobic, chemoheterotrophic, slightly motile short rods with a single flagellum. The temperature, pH and NaCl concentration ranges for growth were 10-37 degrees C, 6.0-9.0 and 0.75-20% (w/v), respectively. Colonies on marine agar were very small (0.3-0.8 mm in diameter), yellowish-brown and very hard. Carotenoid pigments were synthesized but bacteriochlorophyll a was not. Several kinds of pentose, hexose, sugar alcohol, oligosaccharide and amino acid were utilized as sole carbon sources. Oxidase was produced, but catalase was not. All cellular fatty acids were even-numbered monounsaturated or saturated fatty acids and the major fatty acid was cis-7-octadecenoic acid (73.3%). The DNA G + C content of strain HTCC2503T was 60.8 mol%. Phylogenetic analyses of 16S rRNA gene sequences clearly indicated that the strains formed a distinct lineage, allied with activated sludge environmental clone H9, in the alpha-Proteobacteria. The clade containing strains HTCC2503T and HTCC2517 and clone H9 could not be phylogenetically associated with any of the six known orders of the alpha Proteobacteria. From this polyphasic evidence, it is proposed that the novel strains should be classified as Parvularcula bermudensis gen. nov., sp. nov. The type strain is HTCC2503T (= ATCC BAA-594T = KCTC 12087T) and the reference strain is HTCC2517. PMID- 12892123 TI - 'Candidatus phytoplasma ziziphi', a novel phytoplasma taxon associated with jujube witches'-broom disease. AB - Phylogenetic relationships of five jujube witches'-broom (JWB) phytoplasma isolates from four different districts, and other phytoplasmas, were investigated by 16S rDNA PCR amplification and sequence analysis. The 16S rDNA sequences of any pair of the five isolates of JWB phytoplasmas were > 99.5% similar. The JWB phytoplasma 16S rDNA sequences were most closely related to that of the elm yellows (EY) phytoplasma in 16S-group VIII. Phylogenetic analysis of the 16S rDNA sequences from the JWB phytoplasma isolates, together with sequences from most of the phytoplasmas archived in GenBank, produced a tree in which the JWB isolates clustered as a discrete subgroup. The uniqueness of the JWB phytoplasma appears to be correlated with a specific insect vector (Hishimonus sellatus) and the host plant (Zizyphus jujuba), or with a specific geographical distribution. The unique properties of the JWB phytoplasma sequences clearly indicate that it represents a novel taxon, 'Candidatus Phytoplasma ziziphi'. PMID- 12892124 TI - Clostridium caminithermale sp. nov., a slightly halophilic and moderately thermophilic bacterium isolated from an Atlantic deep-sea hydrothermal chimney. AB - A strictly anaerobic, slightly halophilic and moderately thermophilic, sporulating rod designated strain DVird3T was isolated from deep-sea hydrothermal vent samples collected at a depth of approximately 800 m on the Atlantic Ocean Ridge. Strain DVird3T possessed a few laterally inserted flagella, had a DNA G + C content of 33.1 mol% and grew optimally at pH 6.6 and at 45 degrees C. Growth was observed at temperatures between 20 and 58 degrees C and at pH values between 5.8 and 8.2. The optimum NaCl concentration for growth was 3% sea salt (30 g l( 1)); no growth was observed in the presence of 15 or 60 g sea salt l(-1). Strain DVird3T is heterotrophic and utilizes some sugars and various single amino acids. Acetate was the main fatty acid detected from carbohydrate fermentation, together with H2 and CO2. Gelatin was used as an energy source. It performed the Stickland reaction. Phylogenetically, strain DVird3T branched with members of cluster XI of the order Clostridiales, with Clostridium halophilum as its closest relative (similarity of 94.6%). On the basis of its phenotypic, genotypic and phylogenetic characteristics, strain DVird3T (= DSM 15212T = CIP 107654T) is proposed as the type strain of a novel species of the genus Clostridium, Clostridium caminithermale sp. nov. PMID- 12892125 TI - Paenibacillus agarexedens sp. nov., nom. rev., and Paenibacillus agaridevorans sp. nov. AB - Twenty-two agarolytic, aerobic, spore-forming strains were characterized taxonomically by DNA-DNA reassociation experiments, riboprint analyses, 16S rDNA sequencing and phenetic similarity analyses. Based on riboprint analyses, the strains formed eight ribogroups, six of which contained 2-6 strains and two encompassed single strains. Within the multi-strain ribogroups, similarities ranged from 91-99%. Phylogenetic analyses of representatives of the eight groups by 16S rDNA sequence analysis showed that the strains were affiliated to the genus Paenibacillus, but relatedness to described Paenibacillus species was only moderate (<97.8% sequence similarity). Published DNA-DNA similarity values for most of the agarolytic strains, supplemented with new data, supported the distinctiveness of the eight ribogroups. Intragroup DNA-DNA similarity values ranged from 80 to 104%, while intergroup DNA-DNA similarities were <35%. Based on genomic distinctiveness and supported by the presence of distinguishing phenotypic properties, multi-strain groups 1 and 2 are proposed as novel species, Paenibacillus agarexedens sp. nov., nom. rev. (type strain, DSM 1327T=CIP 107437T) and Paenibacillus agaridevorans sp. nov. (type strain, DSM 1355T=CIP 107436T). PMID- 12892126 TI - Halobacillus karajensis sp. nov., a novel moderate halophile. AB - A moderately halophilic, gram-positive, spore-forming bacterium was isolated from surface saline soil of the Karaj region, Iran. The strain, designated MA-2T, was strictly aerobic with rod-shaped cells that occurred singly, in pairs or short chains. It contained L-Om-D-Asp-type peptidoglycan and the major respiratory lipoquinone was MK-7. It was non-motile and had an ellipsoidal endospore located centrally or subterminally. Growth occurred at 10-49 degrees C and in the pH range 6.0-9.6. Strain MA-2T grew at salinities of 1-24% (w/v) NaCl, showing optimal growth at 10% (w/v). The DNA G + C content was 41.3 mol%. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain MA-2T was associated with Bacillus rRNA group 1. The micro-organisms showing the closest phylogenetic relationship to strain MA-2T were Halobacillus litoralis and Halobacillus trueperi. On the basis of phenotypic and chemotaxonomic characteristics, 16S rRNA gene sequence analysis and DNA-DNA similarity data, it is proposed that strain MA 2T (= DSM 14948T = LMG 21515T) should be placed in the genus Halobacillus as the type strain of a novel species, Halobacillus karajensis sp. nov. PMID- 12892127 TI - Corynebacterium atypicum sp. nov., from a human clinical source, does not contain corynomycolic acids. AB - An unusual gram-positive, facultatively anaerobic, catalase-positive, diphtheroid shaped organism originating from an unknown human clinical source was characterized by biochemical, molecular chemical and molecular phylogenetic methods. Based on its morphological and biochemical characteristics and the presence of a murein based on meso-diaminopimelic acid, the unidentified organism was tentatively assigned to the genus Corynebacterium. However, the unknown organism was found to lack the distinctive, short-chain corynomycolic acids that are considered to be characteristic of this genus. Despite the absence of these characteristic lipids, comparative 16S rRNA gene sequencing showed that the unknown bacterium was phylogenetically a member of the genus Corynebacterium and was distinct from all currently known species. Based on both phenotypic and 16S rRNA sequence considerations, it is proposed that the unknown organism be classified as a novel species, Corynebacterium atypicum sp. nov. The type strain of C. atypicum is strain R2070T (=CCUG 45804T=CIP 107431T). PMID- 12892128 TI - Description of Enterococcus canis sp. nov. from dogs and reclassification of Enterococcus porcinus Teixeira et al. 2001 as a junior synonym of Enterococcus villorum Vancanneyt et al 2001. AB - Strains from anal swabs and chronic otitis externa in dogs were shown to be phylogenetically related to the Enterococcus faecium species group. They shared a number of phenotypic characteristics with these species, but they could be easily differentiated by biochemical reactions. In addition, the canine strains were unusual in their nearly complete failure to grow on sodium azide-containing enterococci-selective media and in their Voges-Proskauer reactions (usually negative). By using 16S rRNA sequencing and DNA-DNA hybridization of representative strains, as well as tDNA interspacer gene PCR and SDS-PAGE of whole-cell proteins, the group of canine strains was shown to constitute a novel enterococcal species. The name Enterococcus canis sp. nov. is proposed for this species, with LMG 12316T (= CCUG 46666T) as the type strain. Concurrently, the taxonomic situation and nomenclatural position of Enterococcus porcinus were investigated. As no phenotypic or genotypic differences were found between this species and Enterococcus villorum, the name E. porcinus is considered to be a junior synonym of E. villorum. PMID- 12892129 TI - Classification of Ralstonia pickettii-like isolates from the environment and clinical samples as Ralstonia insidiosa sp. nov. AB - Thirteen Ralstonia pickettii-like isolates from the environment (water, soil and activated sludge) and human clinical samples (including respiratory secretions of cystic fibrosis patients) were investigated in a polyphasic taxonomic study that employed 16S rDNA sequence analysis, DNA-DNA hybridization, determination of DNA base composition, whole-cell protein analysis, biochemical characterization and PCR-based assays. All isolates were classified as a novel Ralstonia species, for which the name Ralstonia insidiosa sp. nov. is proposed. The type strain, LMG 21421T (= CCUG 46789T), was isolated from the sputum of a patient with acute lymphoblastic leukaemia. R. insidiosa can be differentiated from other species of the genus Ralstonia and phenotypically similar species (including the Burkholderia cepacia complex and Achromobacter xylosoxidans) by a variety of biochemical tests, whole-cell protein analysis and several PCR-based assays. Some outstanding issues in the taxonomy of the genus Ralstonia are also discussed. PMID- 12892130 TI - Alicyclobacillus sendaiensis sp. nov., a novel acidophilic, slightly thermophilic species isolated from soil in Sendai, Japan. AB - An acidophilic, slightly thermophilic bacterium, designated strain NTAP-1T, that produces a thermostable extracellular acid collagenase activity with potential industrial applications was isolated from soil of Aoba-yama Park, Sendai, Japan. The temperature range for growth was 40-65 degrees C, with an optimum at 55 degrees C, and the pH range for growth was 2.5-6.5, with an optimum at pH 5.5. Analysis of the 16S rDNA sequence of strain NTAP-1T showed that it is most closely related to strains of the genus Alicyclobacillus. Consistently, the major constituents of the cell-membrane lipid of strain NTAP-1T were omega-alicyclic fatty acids. However, DNA-DNA reassociation studies showed only low similarities (less than 33%) to any type strain of Alicyclobacillus. On the basis of the phenotypic and genotypic properties, a novel species is proposed, Alicyclobacillus sendaiensis sp. nov., represented by strain NTAP-1T (=JCM 11817T=ATCC BAA-609T). PMID- 12892131 TI - Sterolibacterium denitrificans gen. nov., sp. nov., a novel cholesterol oxidizing, denitrifying member of the beta-Proteobacteria. AB - A bacterial strain (Chol-1S(T)) that is able to oxidize cholesterol to CO2 and reduce nitrate to dinitrogen was enriched and isolated from an upflow sludge bed (USB) anoxic reactor that treats sanitary landfill leachate from the city of Montevideo, Uruguay. Cells of strain Chol-1S(T) were gram-negative, rod-shaped to slightly curved, measured 0.5-0.6 x 1.0-1.3 microm and were motile by a single polar flagellum. Strain Chol-1S(T) grew optimally at 30-32 degrees C and pH 7.0, with a doubling time of 44-46 h when cholesterol was used as the sole carbon and energy source. The metabolism of strain Chol-1S(T) was strictly respiratory, with oxygen or nitrate as the terminal electron acceptor. The presence of ubiquinone Q 8 as the sole respiratory lipoquinone indicated that strain Chol-1S(T) belonged to the beta-subclass of the Proteobacteria. Phosphatidylethanolamine was the predominant polar lipid and the G + C content of the DNA was 65.3 mol%. The fatty acid profile of strain Chol-1S(T), cultivated under denitrifying conditions by using a defined mineral medium supplemented with cholesterol, was characterized by the following major components: summed feature 4 (C16:1 omega7c and/or iso C15:0 2-OH), C16:0, C18:1 omega7c and hydroxy acid C10:0 3-OH. Minor components included C10:0, C11:0, C12:0, C14:0, C15:0, C19:0, C19:0 10-methyl and hydroxylated acids C8:0 3-OH and C16:0 3-OH. Analysis of the 16S rDNA sequence showed that strain Chol-1S(T) represents a separate lineage within the Thauera, Azoarcus, Zoogloea and Rhodocyclus assemblage of the beta-Proteobacteria. Strain Chol-1S(T) had highest sequence similarity (96.5%) with strain 72Chol, a denitrifying beta-Proteobacterium. On the basis of polyphasic evidence, strain Chol-1S(T) (=DSM 13999T=ATCC BAA-354T) is proposed as the type strain of Sterolibacterium denitrificans gen. nov., sp. nov. PMID- 12892132 TI - Characterization of several Psychrobacter strains isolated from Antarctic environments and description of Psychrobacter luti sp. nov. and Psychrobacter fozii sp. nov. AB - Eleven psychrophilic bacteria isolated from Antarctic coastal marine environments were subjected to a polyphasic taxonomic study. The isolates were oxidase positive, halotolerant, gram-negative, non-motile coccobacilli with a strictly oxidative metabolism. The DNA G + C content ranged from 44 to 47 mol%. DNA-DNA hybridization experiments showed six homology groups, two of them related at the species level to the type strain of Psychrobacter immobilis, LMG 7203T (70-83%). The highest DNA relatedness of two other groups to known Psychrobacter species was found to the type strain of Psychrobacter glacincola, LMG 21282T (51-57%), and no significant similarity was found between Psychrobacter type strains and the last two groups. The predominant cellular fatty acids detected were typical of the genus Psychrobacter and included 18:1omega9c, 16:1omega7c and 17:1omega8c. 16S rRNA gene sequence analysis confirmed that the strains isolated belonged to the genus Psychrobacter. The results of the study assigned five isolates to P. immobilis, three isolates to P. glacincola and three isolates to novel Psychrobacter species. The names Psychrobacter luti sp. nov. (type strain NF11T=LMG 21276T=CECT 5885T) and Psychrobacter fozii sp. nov. (type strain NF23T=LMG 21280T=CECT 5889T) are proposed for these organisms. PMID- 12892133 TI - Streptococcus oligofermentans sp. nov., a novel oral isolate from caries-free humans. AB - Five streptococcal strains were isolated from dental plaque and saliva of caries free humans. The cells were gram-positive, non-spore-forming, non-motile cocci that were arranged in short chains. The strains were catalase-negative, facultatively anaerobic and produced lactic acid exclusively from glucose fermentation. Biochemical analysis that used both conventional methods and the commercial API 20 Strep system showed that the five strains fermented only a few kinds of sugar. The mean DNA G + C content of the five novel strains was 39.5 +/- 0.8 mol%. Phylogenetic analysis based on 16S rDNA sequence homology indicated that the new isolates represented a novel member of the mitis group of the genus Streptococcus, related most closely to the recently described species Streptococcus sinensis. DNA-DNA relatedness between novel strain LMG 21535T and type strains of phylogenetically related species of oral streptococci was 7.1 16.4%. Therefore a novel Streptococcus species, Streptococcus oligofermentans sp. nov., is proposed. The type strain is LMG 21535T=AS 1.3089T. PMID- 12892134 TI - Assignment of 'Alteromonas marinoglutinosa' NCIMB 1770 to Pseudoalteromonas mariniglutinosa sp. nov., nom. rev., comb. nov. AB - The taxonomic position of the marine bacterium 'Alteromonas marinoglutinosa' NCIMB 1770 was investigated in a polyphasic study. Analysis of 16S rDNA sequence and DNA-DNA reassociation values confirmed the phylogenetic position of strain NCIMB 1770 within the genus Pseudoalteromonas as a separate species, distinct from all Pseudoalteromonas species with validly described names. On the basis of physiological and molecular properties, it is proposed that strain NCIMB 1770 is classified as Pseudoalteromonas mariniglutinosa sp. nov., nom. rev., comb. nov., with the type strain NCIMB 1770T (=KMM 3635T). PMID- 12892135 TI - Rhodocista pekingensis sp. nov., a cyst-forming phototrophic bacterium from a municipal wastewater treatment plant. AB - A novel bacterial species, Rhodocista pekingensis sp. nov., was isolated from a municipal wastewater treatment plant and characterized by polyphasic taxonomy. Cells of R. pekingensis were gram-negative, motile by a single polar flagellum, vibrioid to spiral, 0.6-0.8 microm in width and 0.8-1.5 microm in length. R bodies were not observed. Phototrophically grown cells contained lamellar photosynthetic membranes and bacteriochlorophyll a. Cell growth was anaerobically phototrophic or aerobically chemoheterotrophic. Anaerobically grown cultures were pink-reddish. Thiamin and vitamin B12, but not biotin, were required for growth and 0.05% yeast extract stimulated growth. Acetate, lactate, pyruvate and succinate supported growth. Cysts were formed when butyrate was used as the sole carbon source. Molecular hydrogen (H2), but not sulfide or thiosulfate, was used as an electron donor. The major cellular quinone was Q-9. The DNA G + C content of cells was 68.8 mol%. The type strain of Rhodocista pekingensis is 3-pT (=AS 1.2194T=JCM 11689T). PMID- 12892136 TI - Aurantimonas coralicida gen. nov., sp. nov., the causative agent of white plague type II on Caribbean scleractinian corals. AB - A bacterium previously isolated from a diseased colony of the scleractinian coral Dichocoenia stokesi (common name elliptical star coral) was subjected to a detailed polyphasic taxonomic characterization. The isolate, designated WP1T, was halophilic and strictly aerobic and formed golden-orange-pigmented colonies after prolonged incubation. Cells of WP1T were gram-negative, rod-shaped and showed a characteristic branching rod morphology. Chemotaxonomically, WP1T was characterized by having Q-10 as the major respiratory lipoquinone and sym homospermidine as the main component of the cellular polyamine content. The predominant constituent in the cellular fatty acid profile was C18:1 omega7c, along with C19:0 cyclo omega8c and C16:0. Other fatty acids present in smaller amounts were C17:0, C18:0, C16:1 omega7c, C20:1 omega7c and C18:1 2-OH. The major polar lipids were phosphatidylethanolamine, phosphatidylglycerol and phosphatidylcholine. Minor amounts of diphosphatidylglycerol, phosphatidylmonomethylethanolamine and phosphatidyldimethylethanolamine were present. The G + C content of the genomic DNA was 66.3 mol%. Phylogenetic analysis of the 16S rRNA gene sequence showed that WP1T represents a separate subline of descent within the order 'Rhizobiales' of the 'Alphaproteobacteria'. The new line of descent falls within the group of families that includes the Rhizobiaceae, Bartonellaceae, Brucellaceae and 'Phyllobacteriaceae', with no particular relative within this group. The 16S rRNA gene sequence similarity to all established taxa within this group was not higher than 92.0% (to Mesorhizobium mediterraneum). To accommodate this emerging coral pathogen, the creation of a new genus and species is proposed, Aurantimonas coralicida gen. nov., sp. nov. (type strain WP1T = CIP 107386T = DSM 14790T). PMID- 12892137 TI - Leuconostoc inhae sp. nov., a lactic acid bacterium isolated from kimchi. AB - Six strains of a hitherto unknown bacterium isolated from kimchi, a fermented vegetable food produced in Korea, were characterized by using phenotypic methods, phylogenetic analysis and DNA-DNA hybridization. The novel strains were gram positive, non-spore-forming, heterofermentative and spherical or lenticular lactic acid bacteria. Comparative 16S rRNA gene sequencing and DNA relatedness demonstrated that the unknown strains represented a novel clade within the genus Leuconostoc and were close to, but distinct from, Leuconostoc gelidum. The unknown strains were clearly distinguished from all described members of the genus Leuconostoc by using RFLP pattems of genus-specific 16S rRNA gene PCR products with a single endonuclease, BsmAI. Based on the polyphasic evidence, the unknown isolates are classified as Leuconostoc inhae sp. nov. The type strain is strain IH003T (= KCTC 3774T = DSM 15101T). PMID- 12892138 TI - Reclassification of Desulfobacterium macestii as Desulfomicrobium macestii comb. nov. AB - Phylogenetic, chemotaxonomic and metabolic data obtained for Desulfobacterium macestii indicate that this species is not a member of the genus Desulfobacterium, but of the genus Desulfomicrobium. Phylogenetically, it is closely related to Desulfomicrobium baculatum and Desulfomicrobium norvegicum, but it can be differentiated from these species by its metabolic properties. It is therefore proposed to reclassify Desulfobacterium macestii as Desulfomicrobium macestii comb. nov. PMID- 12892139 TI - Tepidibacter thalassicus gen. nov., sp. nov., a novel moderately thermophilic, anaerobic, fermentative bacterium from a deep-sea hydrothermal vent. AB - A moderately thermophilic, anaerobic, endospore-forming bacterium (strain SC 562T) was isolated from a hydrothermal vent chimney located at 13 degrees N on the East-Pacific Rise at a depth of 2650 m. Cells of strain SC 562T were straight to slightly curved rods, which were 0.7-0.9 microm in diameter and 3.5-6.0 microm in length with peritrichous flagella. Strain SC 562T formed round, refractile endospores in terminally swollen sporangia. The temperature range for growth was 33-60 degrees C, with an optimum at 50 degrees C. The pH range for growth was 4.8 8.5, with an optimum at pH 6.5-6.8. Growth of strain SC 562T was observed at NaCl concentrations ranging from 1.5 to 6% (w/v). The substrates utilized by strain SC 562T included casein, peptone, albumin, yeast extract, beef extract, alanine plus proline and starch. Glucose, maltose, pyruvate, valine and arginine each slightly stimulated growth in the presence of yeast extract. The products of glucose fermentation were ethanol, acetate, H2 and CO2. Strain SC 562T reduced elemental sulfur to hydrogen sulfide. The G + C content of the DNA of strain SC 562T was 24 mol%. 16S rDNA sequence analysis revealed that the isolated organism belonged to cluster XI of the Clostridium subphylum. On the basis of its physiological properties and phylogenetic analyses, it is proposed that strain SC 562T represents the sole species of a novel genus, Tepidibacter; the name Tepidibacter thalassicus is proposed for strain SC 562T (=DSM 15285T=UNIQEM 215T). PMID- 12892140 TI - Isolation of Corynebacterium falsenii and description of Corynebacterium aquilae sp. nov., from eagles. AB - Biochemical, molecular chemical and molecular genetic studies were performed on seven unidentified gram-positive, rod-shaped organisms recovered from eagles. The strains were provisionally identified as Corynebacterium jeikeium with the commercial API Coryne system, but they were able to grow under anaerobic conditions and were non-lipophilic. Comparative 16S rRNA gene sequencing studies demonstrated that the isolates belonged phylogenetically to the genus Corynebacterium. Three strains were identified genotypically as Corynebacterium falsenii; the remaining four strains corresponded to a hitherto unknown lineage within the genus Corynebacterium, associated with a small subcluster of species that included Corynebacterium diphtheriae and its close relatives. The unknown bacterial strains were readily distinguished from these and other species of the genus by biochemical tests. Based on both phenotypic and phylogenetic evidence, it is proposed that the unknown bacterial strains from eagles should be classified as Corynebacterium aquilae sp. nov. (type strain is S-613T = CECT 5993T = CCUG 46511T). PMID- 12892141 TI - Dechlorosoma suillum Achenbach et al. 2001 is a later subjective synonym of Azospira oryzae Reinhold-Hurek and Hurek 2000. AB - In order to clarify the taxonomic position of Dechlorosoma suillum, which shares 99.9% 16S rDNA sequence identity (1433 of 1435 bp) with Azospira oryzae, we compared the two species in a polyphasic taxonomic approach. Results of 122 physiological and biochemical tests for D. suillum DSM 13638T and Azospira oryzae 6a3T were identical, except for the lack of growth of Azospira oryzae 6a3T with perchlorate as the terminal electron acceptor. Presence of a nifH gene and nitrogenase activity, a key feature of Azospira, were also detected in D. suillum by Southern hybridization and by the acetylene reduction assay, respectively. Whole-cell SDS-PAGE profiles of SDS-soluble proteins of strains DSM 13638T and 6a3T were almost identical. DNA-DNA hybridization studies showed more than 90% binding between D. suillum and two strains of Azospira oryzae. These data provide evidence that the two bacteria belong to the same species and that D. suillum is a later subjective synonym of Azospira oryzae. PMID- 12892142 TI - Vulcanithermus mediatlanticus gen. nov., sp. nov., a novel member of the family Thermaceae from a deep-sea hot vent. AB - A novel thermophilic, microaerophilic, facultatively chemolithoheterotrophic bacterium designated strain TR(T) was isolated from a sample of a deep-sea hydrothermal chimney collected at the Rainbow vent field on the Mid-Atlantic Ridge (36 degrees 14'N). Gram-negative, non-spore-forming, non-motile rods occurred singly or in pairs. The organism grew in the temperature range 37-80 degrees C with an optimum at 70 degrees C and at pH 5.5-8.4 with an optimum around 6.7. The NaCl range for growth was 10-50 g l(-1) with an optimum of 30 g l(-1). Strain TR(T) grew chemoorganoheterotrophically with carbohydrates, proteinaceous substrates, organic acids and alcohols using oxygen or nitrate as electron acceptors. The isolate was able to grow at oxygen concentrations from 0.5 to 21%. Oxygen concentrations that promoted fastest growth ranged from 4 to 8% under agitation. The novel isolate was able to grow lithoheterotrophically with molecular hydrogen as the energy source. The G + C content of the genomic DNA was 68.4 mol%. Phylogenetic analysis of the 16S rDNA sequence placed strain TR(T) within the phylum Deinococcus-Thermus of the Bacteria. On the basis of phenotypic and phylogenetic data, it is proposed that this isolate should be described as a member of a novel species of a new genus as Vulcanithermus mediatlanticus gen. nov., sp. nov. The type strain is TR(T) (= DSM 14978T = VKM B 2292T = JCM 11956T). PMID- 12892143 TI - Caldisphaera lagunensis gen. nov., sp. nov., a novel thermoacidophilic crenarchaeote isolated from a hot spring at Mt Maquiling, Philippines. AB - Four novel, thermoacidophilic, crenarchaeotic cocci that grew anaerobically and heterotrophically were isolated from an acidic hot spring in the Philippines; two representative strains were characterized in detail. Most cells were regular cocci, 0.8-1.1 microm in width, which occurred singly or in pairs. They were non motile and grew at 45-80 degrees C (optimum 70-75 degrees C) and pH 2.3-5.4 (optimum 3.5-4.0). They utilized starch, glycogen, gelatin, beef extract, yeast extract and peptone as carbon and energy sources. Growth was stimulated by the presence of sulfur as an electron acceptor. The lipid fraction contained cyclic and acyclic tetraether core lipids. The DNA G + C content was 31 mol%; phylogenetic analysis based on 16S rDNA sequences showed that the novel cocci represent an independent lineage in the phylum Crenarchaeota, distantly related to Acidilobus aceticus and an allied strain, NC12. Caldisphaera lagunensis gen. nov., sp. nov. is proposed to accommodate the four strains. The type strain is IC 154T (=JCM 11604T=MCC-UPLB 1331T=ANMR 0165T). PMID- 12892144 TI - Gemmatimonas aurantiaca gen. nov., sp. nov., a gram-negative, aerobic, polyphosphate-accumulating micro-organism, the first cultured representative of the new bacterial phylum Gemmatimonadetes phyl. nov. AB - A phylogenetically novel aerobic bacterium was isolated from an anaerobic-aerobic sequential batch reactor operated under enhanced biological phosphorus removal conditions for wastewater treatment. The isolation strategy used targeted slowly growing polyphosphate-accumulating bacteria by combining low-speed centrifugations and prolonged incubation on a low-nutrient medium. The isolate, designated strain T-27T, was a gram-negative, rod-shaped aerobe. Cells often appeared to divide by budding replication. Strain T-27T grew at 25-35 degrees C with an optimum growth temperature of 30 degrees C, whilst no growth was observed below 20 degrees C or above 37 degrees C within 20 days incubation. The pH range for growth was 6.5-9.5, with an optimum at pH 7.0. Strain T-27T was able to utilize a limited range of substrates, such as yeast extract, polypepton, succinate, acetate, gelatin and benzoate. Neisser staining was positive and 4,6 diamidino-2-phenylindole-stained cells displayed a yellow fluorescence, indicative of polyphosphate inclusions. Menaquinone 9 was the major respiratory quinone. The cellular fatty acids of the strain were mainly composed of iso C15:0, C16:1 and C14:0. The G + C content of the genomic DNA was 66 mol%. Comparative analyses of 16S rRNA gene sequences indicated that strain T-27T belongs to candidate division BD (also called KS-B), a phylum-level lineage in the bacterial domain, to date comprised exclusively of environmental 16S rDNA clone sequences. Here, a new genus and species are proposed, Gemmatimonas aurantiaca (type strain T-27T=JCM 11422T=DSM 14586T) gen. nov., sp. nov., the first cultivated representative of the Gemmatimonadetes phyl. nov. Environmental sequence data indicate that this phylum is widespread in nature and has a phylogenetic breadth (19% 16S rDNA sequence divergence) that is greater than well known phyla such as the Actinobacteria (18% divergence). PMID- 12892145 TI - Schlegelella thermodepolymerans gen. nov., sp. nov., a novel thermophilic bacterium that degrades poly(3-hydroxybutyrate-co-3-mercaptopropionate). AB - A novel thermophilic bacterium, strain K14T, capable of degrading poly(3 hydroxybutyrate) as well as copolymers containing 3-hydroxybutyrate and 3 mercaptopropionate linked by thioester bonds, was isolated. 16S rDNA sequence analysis showed that strain DhA-71, a dehydroabietic acid-degrading bacterium, was the nearest phylogenetic neighbour and that both strains should be placed as members of a newly created genus, Schlegelella gen. nov., in the Rubrivivax subgroup of the beta-Proteobacteria. Strain K14T (= LMG 21644T = DSM 15344T) is proposed as the type strain of Schlegelella thermodepolymerans gen. nov., sp. nov. Its phylogenetic, morphological, biochemical and chemotaxonomic characteristics are described in detail. PMID- 12892146 TI - Erythrobacter flavus sp. nov., a slight halophile from the East Sea in Korea. AB - Two gram-negative, motile, non-spore-forming, yellow-pigmented and slightly halophilic strains (SW-46T and SW-52) were isolated from sea water of the East Sea, Korea, and subjected to a polyphasic taxonomic study. Strains SW-46T and SW 52 were characterized chemotaxonomically by having ubiquinone-10 (Q-10) as the predominant respiratory lipoquinone and C18:1 omega7c as the major fatty acid. Their DNA G + C content was 64.0-64.1 mol%. Strains SW-46T and SW-52 showed 1 bp difference in their 16S rDNA sequences and a mean DNA-DNA relatedness level of 94.4%. Phylogenetic analysis based on 16S rDNA sequences showed that strains SW 46T and SW-52 fall within the alpha-subclass of the Proteobacteria and form a coherent cluster with Erythrobacter longus, Erythrobacter litoralis and Erythrobacter citreus. Levels of 16S rDNA similarity between strains SW-46T and SW-52 and the type strains of these three Erythrobacter species were 96.5-97.9%. Levels of DNA-DNA relatedness between strains SW-46T and SW-52 and the type strains of E. longus, E. litoralis and E. citreus were 3.6-14.7%. Therefore, on the basis of phenotypic properties, phylogeny and genomic data, strains SW-46T and SW-52 should be placed in the genus Erythrobacter as a novel species, for which the name Erythrobacter flavus sp. nov. is proposed. The type strain is SW 46T (= KCCM 41642T = JCM 11808T). PMID- 12892147 TI - Phylogeny of the photosynthetic euglenophytes inferred from the nuclear SSU and partial LSU rDNA. AB - Previous studies using the nuclear SSU rDNA have indicated that the photosynthetic euglenoids are a monophyletic group; however, some of the genera within the photosynthetic lineage are not monophyletic. To test these results further, evolutionary relationships among the photosynthetic genera were investigated by obtaining partial LSU nuclear rDNA sequences. Taxa from each of the external clades of the SSU rDNA-based phylogeny were chosen to create a combined dataset and to compare the individual LSU and SSU rDNA datasets. Conserved areas of the aligned sequences for both the LSU and SSU rDNA were used to generate parsimony, log-det, maximum-likelihood and Bayesian trees. The SSU and LSU rDNA consistently generated the same seven terminal clades; however, the relationship among those clades varied depending on the type of analysis and the dataset used. The combined dataset generated a more robust phylogeny, but the relationships among clades still varied. The addition of the LSU rDNA dataset to the euglenophyte phylogeny supports the view that the genera Euglena, Lepocinclis and Phacus are not monophyletic and substantiates the existence of several well supported clades. A secondary structural model for the D2 region of the LSU rDNA was proposed on the basis of compensatory base changes found in the alignment. PMID- 12892148 TI - Three new combinations from the Cryptococcus laurentii complex: Cryptococcus aureus, Cryptococcus carnescens and Cryptococcus peneaus. AB - Fifteen strains of the Cryptococcus laurentii complex were reclassified based on sequence analyses of 18S rDNA, the D1/D2 region of the 26S rDNA and the internal transcribed spacer regions, as well as physiological and biochemical properties. The strains were divided into phylogenetic groups I and II. The type strain of C. laurentii (CBS 139T=ATCC 18803T=JCM 9066T=MUCL 30398T=NRRL Y-2536T) was in phylogenetic group I. Phylogenetic group II, which was phylogenetically distant from phylogenetic group I, clustered with Cryptococcus dimennae and Bullera globispora. In phylogenetic group I, the type strain of Torula aurea (CBS 318T=ATCC 32063T=IFO 0372T=NRRL Y-1582T) appeared to be a separate species from C. laurentii, and the designation Cryptococcus aureus comb. nov. is proposed for Torula aurea. Cryptococcus flavescens (formerly Torula flavescens, type strain CBS 942T=ATCC 10668T=DBVPG 6007T=MUCL 30414T) was treated as a synonym of C. laurentii; however, this is a distinct species. The type strains of Torulopsis carnescens (CBS 973T=ATCC 32064T=MUCL 30641T=NRRL Y-1503T), Rhodotorula peneaus (CBS 2409T=ATCC 13546T=MUCL 30643T=NRRL Y-2005T) and Cryptococcus victoriae belonged to phylogenetic group II. Two new combinations, Cryptococcus carnescens comb. nov. (type strain CBS 973T=ATCC 32064T=MUCL 30641T=NRRL Y-1503T) and Cryptococcus peneaus comb. nov. (type strain CBS 2409T=ATCC 13546T=MUCL 30643T=NRRL Y-2005T), are proposed from this group. PMID- 12892149 TI - Emended phenotypic characterization of Prototheca zopfii: a proposal for three biotypes and standards for their identification. AB - A representative selection of Prototheca zopfii strains isolated from different environmental habitats or clinical cases was characterized in a polyphasic approach in order to assess their intraspecies taxonomic position. Recently, the recognition of distinct phenotypic clusters has been reported as the assignment of 'variants'. In the present study, 11 strains were compared by a number of phenotypic and genetic criteria, including growth characteristics, biochemical reactions and serotyping results. Based on emended standards for biotype identification, P. zopfii strains showed auxanographic differences and distinct assimilation patterns with respect to utilization of amino acids and glycerol. Serotyping by means of immunoblotting revealed that all isolates of variant II obtained from clinical cases, i.e. isolates from bovine mastitis or from human enteropathia, showed specific antigen patterns. They were found to be different from strains assigned to the other two variants with respect to their immunogenic antigens. Furthermore, comparison of partial 18S rDNA sequences confirmed distinct differences between the former variants. Based on these results, it is proposed that P. zopfii merits classification as a species comprising three biotypes. PMID- 12892150 TI - Phylogenetic analysis identifies the 'megabacterium' of birds as a novel anamorphic ascomycetous yeast, Macrorhabdus ornithogaster gen. nov., sp. nov. AB - An organism commonly referred to as 'megabacterium' colonizes the gastric isthmus of many species of birds. It is weakly gram-positive and periodic acid-Schiff positive and stains with silver stains. Previous studies have shown that it has a nucleus and a cell wall similar to those seen in fungi. Calcofluor white M2R staining suggests that the cell wall contains chitin, a eukaryote-specific substance, and rRNA in situ hybridization demonstrates that it is a eukaryote. To characterize this organism phylogenetically, DNA was extracted from purified cells. rDNA was readily amplified by PCR with pan-fungal DNA primer sets and primer sets derived from the newly determined sequence, but not with bacteria specific primer sets. Specific primer sets amplified rDNA from isthmus scrapings from an infected bird, but not from a non-infected bird or other control DNA. The sequence was confirmed to derive from the purified organism by in situ rRNA hybridization using a specific probe. Phylogenetic analysis of sequences of the 18S rDNA and domain D1/D2 of 26S rDNA showed the organism to be a previously undescribed anamorphic ascomycetous yeast representing a new genus. The name Macrorhabdus ornithogaster gen. nov., sp. nov. is proposed for this organism. The type material is CBS 9251T (= NRRL Y-27487T). PMID- 12892151 TI - Description of new Ensifer strains from nodules and proposal to transfer Ensifer adhaerens Casida 1982 to Sinorhizobium as Sinorhizobium adhaerens comb. nov. Request for an opinion. AB - A group of four diverse rhizobial isolates and two soil isolates that are highly related to Ensifer adhaerens were characterized by a polyphasic approach. On the basis of DNA-DNA hybridizations and phenotypic features, these strains cannot be distinguished clearly form Ensifer adhaerens, a soil bacterium that was described in 1982, mainly on the basis of phenotypic characteristics. Phylogenetically, Ensifer and Sinorhizobium form a single group in the 16S rDNA dendrogram of the alpha-Proteobacteria, as well as in an analysis of partial recA gene sequences. They may therefore be regarded as a single genus. Because Sinorhizobium was proposed in 1988, according to the Bacteriological Code (1990 Revision) the older name, Ensifer, has priority. However, there are several reasons why a change from Sinorhizobium to Ensifer may not be the best solution and making an exception to Rule 38 may be more appropriate. We therefore propose the species Sinorhizobium adhaerens comb. nov. and put forward a Request for an Opinion to the Judicial Commission regarding the conservation of Sinorhizobium adhaerens over Ensifer adhaerens. PMID- 12892152 TI - Why do we help a micropreemie to live? AB - Helping a "micropreemie" to live by aggressive interventions may sometimes seem unnatural. However, utilitarian assessment of benefits derived from lifesaving efforts for a micropreemie is considered inappropriate. The goal in treating premature infants has advanced from fetal salvage to achieving "intact survival", which represents a new therapeutic target. In this way, the record for lifesaving in extremely low-birthweight infants is continually being broken. Why do we help a micropreemie to live? Moral and ethical emotions are the underlying reasons for the aggressive care devoted to premature infants, including micropreemies. Such human feelings might even be considered the purpose of life. Human emotion is the impetus for aggressive efforts to improve the survival prospects of premature infants. The beautiful and delicate nature of a newborn is compelling. The high order emotion of empathy for another's misfortune is also important. Most human emotions are related to an awareness of death, and micropreemies are near death. In Oriental thought, a human being is a growing product of nature. Forces of nature and changes in a living being follow nature's rules. First and foremost, an individual life is part of the long chain of existence beginning before the self and continuing beyond it. An immature human being, even a newborn, is simultaneously a complete entity and part of nature's long chain of being, which has a wholeness of its own that affirms a micropreemie's right to life. A fetus is a member of human society in the sense that there is an overall reverence for life as a quality that lives on. CONCLUSION: The limit that bioethics must not exceed is the sanctity of life. We believe that the birth of a micropreemie is an important and serious event. We profoundly wish that a micropreemie might live and thrive, because we on earth must live with the continual presence and imminence of death. PMID- 12892153 TI - "Maturational lag" hypothesis of attention deficit hyperactivity disorder: an update. AB - This article discusses the relationship of maturation to ADHD and hyperkinetic disorder (HKD), with an emphasis on current research in ADHD and HKD, persistence and remission of ADHD symptoms over time and brain maturational trajectories. CONCLUSION: ADHD is a broad, heterogeneous syndrome and only a subgroup of subjects has a diagnosis of HKD, which is a subset of individuals with severe ADHD combined subtype. Children showing symptoms above the threshold for a diagnosis of ADHD are at risk of developing comorbid conditions and increasing stress in both parents and teachers. In some subjects, ADHD symptoms can improve over time during maturation and development. These children with a diagnosis of ADHD could be viewed as showing variants of normal childhood behaviour with maturational trajectories that are lagging behind but will catch up. ADHD could therefore represent a continuum from normality at one extreme to a severe disorder, HKD according to ICD-10, at the other extreme. PMID- 12892154 TI - Compliance of present recommendations of fatty acids in formulas for term infants with the actual human milk fatty acid composition in different populations. AB - Fats are the only dietary agents possibly modulating the individual membrane composition, which otherwise is "genetically" determined. In infancy, human milk represents the first natural source of fats. Although the fat intake of breastfed infants is quantitatively and qualitatively unique, it is not known to what extent their varying requirements are met by the fat composition in the milk of their mothers. CONCLUSION: The fat composition of arbitrarily chosen human milk is not always the best standard for that in infant formula. PMID- 12892155 TI - The challenges of fostering infants and children. AB - There are various reasons for and ways of fostering infants and children in different parts of the world. In a traditional society, for example among Australian Aboriginals, children who have been subjected to catastrophes such as maternal death are taken care of in their extended families, whereas children in the Western world are fostered mainly because of adverse psychosocial factors such as abuse or neglect. In transitional societies and urban slums, where there are few protective mechanisms, the sufferings of orphans may be extreme and they run a great risk of being exploited in various ways or of becoming involved in criminal street gangs. CONCLUSION: The fostering of infants and children around the world creates many yet unanswered challenges. This rather neglected area is in urgent need of much more attention, especially in transitional societies. PMID- 12892156 TI - Fatty acids in formulae for term infants: compliance of present recommendations with the actual human milk fatty acid composition of geographically different populations. AB - AIM: Recommendations for formula fatty acids (FA) are largely based on the mature human milk FA composition. This study aimed to investigate whether current recommendations for formula FA for term infants comply with the actual breast milk FA composition of geographically distinct populations and to provide more realistic grounds for future recommendations. METHODS: 455 mature breast-milk samples were collected in different countries over 25 y. Recommendations of different organizations were projected on their FA data. FA interrelationships were calculated with Spearman's rank tests. FA compositions of 30 formulae were compared with those of breast milk. RESULTS: Many samples from non-Western communities did not meet the recommendations for formula 12:0, 14:0 and 18:2omega6, since these are mainly based on breast milk of mothers living in Western countries. Recommendations for 18:3omega3, 18:2omega6/18:3omega3, 20:4omega6 and 22:6omega3 were not met by many milk samples, which may point to the poorly developed recommendations for long-chain polyunsaturated FA. Most of the investigated breast-milk FA (12:0, 14:0, 16:0, 18:0, 18:3omega3, 22:6omega3, 18:2omega6, 20:4omega6, 18:1omega9) were either positively or negatively interrelated. Many formulae had FA compositions that were not consistent with the physiological interrelationships of FA in breast milk. CONCLUSION: Future recommendations, if based on human milk, should derive from its FA balance, as indicated by the FA interrelationships. A "humanized" formula FA composition would in this sense be any composition that cannot be distinguished from that of breast milk by techniques such as principal component analysis. PMID- 12892157 TI - Vitamin D supplementation to healthy children does not affect serum osteocalcin or markers of type I collagen turnover. AB - AIM: New serum markers have recently been introduced in the assessment of bone turnover. Such measures are osteocalcin, the C-terminal propeptide of type I procollagen (PICP), the N-terminal propeptide of type I procollagen (PINP) and the C-terminal pyridinoline cross-linked telopeptide of type I collagen (ICTP). This study aimed to determine whether supplementation with vitamin D3 to healthy children during the winter affects bone turnover in healthy children measured by serum osteocalcin, PICP, PINP or ICTP. METHODS: 12 girls and 8 boys aged 6.2-13.7 (mean 9.8) y, all proven healthy by medical examination and history, were enrolled in a double-blind, randomized, placebo-controlled, cross-over study with two 4 wk treatment periods and 2 wk washout. Vitamin D3 600 IU was given in one tablet of ABCDin daily. On the last day of the 4 wk periods blood was sampled for assessment of serum osteocalcin, PICP, PINP, ICTP, 25-OH-vitamin D, 1,25-diOH vitamin D and parathyroid hormone (PTH). RESULTS: During supplementation and placebo periods serum osteocalcin (mean +/- SEM) was 53.9 +/- 5.7 and 54.4 +/- 3.8 microg l(-1) (p = 0.70), PICP was 437+/- 44 and 429 +/- 41 microg l(-1) (p = 0.73), PINP was 579 +/- 56 and 619 +/- 64 microg l(-1) (p = 0.33) and ICTP was 13.4 +/- 0.9 and 13.6 +/- 0.7 microg l(-1) (p = 0.52), respectively. Mean +/- SEM serum 25-OH-vitamin D was 47.0 +/- 2.3 and 33.0 +/- 3.0 nmol l(-1) during vitamin D3 supplementation and placebo (p < 0.001, t = 8.10, 95% CI = 10.3 to 17.6 nmol l(-1)), 1,25-diOH-vitamin D and PTH were 87.5 +/- 4.3 and 92.0 +/- 5.3 pmol l(-1) (p = 0.38), and 3.97 +/- 0.5 and 4.21 +/- 0.4 micromol l(-1) (p = 0.37), respectively. CONCLUSION: Supplementation with 600 IU vitamin D3 to healthy children in the winter does not affect bone turnover as measured by serum osteocalcin, PICP, PINP or ICTP. Vitamin D supplementation to healthy children may not be recommended on the ground of concern for bone turnover. PMID- 12892159 TI - Disaccharidase activities in Belgian children: reference intervals and comparison with non-Belgian Caucasian children. AB - AIM: To establish reference values for disaccharidase activities in Belgian children and to compare enzyme activities with those of non-Belgian Caucasian children. METHODS: Data from Belgian children who had undergone endoscopic jejunal biopsies (1994-2000) for suspected malabsorption were reviewed. The patients were divided into three groups based on histology: (A) normal (n = 201), (B) moderate changes (n = 58) and (C) (sub)total atrophy (n = 14). The 95% reference limits for disaccharidase activities (U/g protein) were calculated for group A after exclusion of patients with a positive hydrogen breath test, a history of lactose intolerance or coeliac disease (final population: n = 151, 0.1 12 y). Values were compared with those of 34 non-Belgian Caucasian children with normal histology (28 of Mediterranean origin). RESULTS: The reference limits (90% confidence interval) were 86 (65-111)-423 (366-494) for maltase, 9 (6-12)-91 (78 122) for lactase and 24 (18-30)-155 (120-184) for sucrase. No gender-related differences in enzyme activities were found. Lactase levels showed a slight decrease with increasing age. Disaccharidase activities of children with histologically confirmed mucosal injury were significantly lower than those of children with normal histology: median values for groups A, B and C were 208, 181 and 96, respectively, for maltase, 40, 28 and 7, respectively, for lactase and 69, 54 and 25, respectively, for sucrase. Median disaccharidase activities in biopsies with normal histology were lower in non-Belgian children, the difference being only statistically significant for lactase, 33 versus 40. CONCLUSION: The reference values for Belgian children are well in line with other reported values from Caucasian children. Although enzyme activities are lower in children with histologically confirmed mucosal damage, they do not allow differentiation between histology groups. Lower lactase values were found in non-Belgian children. PMID- 12892158 TI - Weekly oral vitamin K prophylaxis in Denmark. AB - AIM: To evaluate oral vitamin K prophylaxis at birth by giving 2 mg phytomenadione, followed by weekly oral vitamin K prophylaxis; 1 mg was administered by the parents until 3 mo of age. METHODS: A total of 507850 live babies were born in Denmark during the study period, November 1992 to June 2000. Of these infants, 78% and 22% received oral and intra-muscular prophylaxis, respectively; i.e. about 396000 neonates received oral prophylaxis at birth. Weekly oral prophylaxis was recommended for all infants as long as they were mainly breastfed. A survey of possible cases of vitamin K deficiency bleeding (VKDB) was carried out by repeated questionnaires to all Danish paediatric departments and by checking the National Patient Register. RESULTS: No cases of VKDB were revealed, i.e. the incidence was 0-0.9:100000 (95% CI). The questionnaires were used to evaluate compliance with the regimen. Parents of 274 infants participated. A dose of vitamin K was regarded as having been given if the infant received a drop of vitamin K or was mostly formula-fed that week, and the prophylaxis was regarded as completed if the infant had received at least 9 doses. Compliance was good, with 94% of the infants completing the course of prophylaxis. CONCLUSION: Weekly oral vitamin K supplementation during the first 3 mo of life was an efficient prophylaxis against VKBD. Parental compliance with the regimen was good. PMID- 12892160 TI - Psychosocial distress during pregnancy and the risk of infantile colic: a follow up study. AB - AIM: To examine the association between psychosocial exposures during pregnancy and the risk of infantile colic. METHODS: The study included 378 infants and was conducted as a substudy of the Danish National Birth Cohort from 1997 to 1999, with prenatal data collected twice during pregnancy. A diary with a record for postpartum weeks 4-8 was used to quantify the amount of the infants' crying and fussing. RESULTS: The cumulative incidence proportion of infantile colic was 8.2%. A threefold increased risk of infantile colic (OR = 3.7; 95% CI: 1.1-13.2) was found for mothers who reported distress during pregnancy. Close to a twofold increased risk of IC was found for the women who scored higher than 8 on the psychological distress scale (adjusted OR = 1.9; 95% CI: 0.5-7.2). CONCLUSION: The results indicate that general distress during pregnancy influences the risk of infantile colic. Whether or not this relationship is causal requires further investigations. PMID- 12892161 TI - Thymic size in preterm neonates: a sonographic study. AB - AIM: To assess the variation in size of the thymus in vivo in preterm neonates and to identify relations between thymic size and gestational age (GA), birthweight, occurrence of postnatal infections and maternal alcohol and tobacco intake during pregnancy. METHODS: Eighty preterm neonates with a GA between 24 and 36 wk, and a birthweight between 490 and 4110 g were examined between days 0 and 19 after birth. The thymic size was assessed by sonography as a volume estimate, the so-called thymic index (Ti). The median Ti was 5.2 (1.2-17.9). Ti was positively correlated with birthweight and GA and negatively correlated with occurrence of postnatal infection (p < 0.01, p = 0.03, p = 0.05, R2 = 0.68). A correlation between thymic size and maternal alcohol and tobacco intake was not demonstrated. CONCLUSION: It is possible to assess the size of the thymus by sonography in very low-birthweight and preterm neonates. A normal range for Ti in preterm neonates has been established. The sonographic method is a safe and effective technique for measuring the size of the thymus in preterm infants. PMID- 12892162 TI - Early use of Nasal-BiPAP in two infants with Congenital Central Hypoventilation syndrome. AB - AIM: To reduce the problems caused by prolonged artificial ventilation in babies with Congenital Central Hypoventilation syndrome (CCHS). METHODS: Two term infants with CCHS, weighing 4030 g and 3100 g, respectively, at the beginning of treatment and aged 53 and 31 d, respectively, were successfully ventilated with a Nasal Bilevel Positive Airway Pressure (N-BiPAP) device. RESULTS: In the first patient the tcPO2 recordings (mean +/- SD) during sleep were 46 +/- 12 mmHg before using N-BiPAP and 58 +/- 13 mmHg after using the device, while those for tcPCO2 were 75 +/- 9 mmHg and 49 +/- 11 mmHg, respectively. In the second patient tcPO2 during sleep was 42 +/- 3 mmHg before, and 55 +/- 5 after N-BiPAP, and for tcPCO2 the recordings were 119 +/- 24 mmHg and 55 +/- 6 mmHg, respectively, showing a significant improvement. One infant had persistent gastro-oesophageal reflux, and frontal skin abrasion caused by the face mask. Nevertheless, these complications did not necessitate the discontinuation of N-BiPAP ventilation, thus precluding prolonged use of intubation and tracheotomy. CONCLUSION: In infants with CCHS, early use of non-invasive, positive-pressure ventilation with N-BiPAP, in association with careful monitoring, can decrease problems caused by prolonged intubation and tracheotomy. PMID- 12892163 TI - Short- and long-term effects of neonatal glucocorticoid therapy: is hydrocortisone an alternative to dexamethasone? AB - AIM: To compare short-term effects and neurodevelopmental outcome of neonatal glucocorticoid therapy between two centres. METHODS: A retrospective study was performed in two centres using a tapering course of either 5 to 1 mg kg(-1) hydrocortisone (HC; 22 d) or 0.5 to 0.1 mg kg(-1) dexamethasone (DEX; 21 d). In both centres glucocorticoid-treated infants and control patients were matched for gestational age, birthweight, severity of infant respiratory distress syndrome and periventricular-intraventricular haemorrhage. The following short-term glucocorticoid-induced effects were investigated in 25 HC-treated and 25 control patients in centre A, and in 23 DEX-treated and 23 control patients in centre B: oxygen dependency (inspiratory oxygen fraction), arterial pressure, blood glucose and urea concentrations, weight gain and head circumference before, during and after therapy (in treated infants), or at an interval comparable to treated infants (in control infants). Neurological outcome, psychomotor development and school performance at 5-7 y of age was evaluated in all groups. RESULTS: HC and DEX were equally potent in reducing oxygen dependency. Mean arterial pressure as well as blood glucose and urea concentrations were significantly increased during DEX, but not during HC treatment. Weight gain stopped during DEX therapy, but not during HC. Head circumference in both treatment groups was decreased after therapy compared with controls. Neonatally DEX-treated children needed special school education significantly more often (p < 0.01) than controls at 5-7 y of age. No differences between neonatally HC-treated children and controls on neurodevelopmental outcome were found at 5-7 y of age. CONCLUSION: Neonatal HC therapy has fewer short- and long-term adverse effects than neonatal DEX therapy. PMID- 12892164 TI - Hypercontractile heart failure caused by catecholamine therapy in premature neonates. AB - Catecholamine treatment for hypotension is common practice in neonatal intensive care units. In the presence of left ventricular hypertrophy, the positive inotropic action of catecholamines with mainly beta-receptor specificity can lead to excessive hypercontractility and paradoxical hypotension. This reports presents two cases of hypercontractile heart failure during beta-agonist treatment in very low birthweight (VLBW) infants. Both patients (27 wk, 5 d; and 26 wk, 6 d of gestation) underwent surgical arterial duct ligation and coarctation repair. After operation they developed arterial hypotension that failed to respond to epinephrine (adrenaline) or dobutamine treatment. Echocardiography demonstrated a complete systolic obliteration of the left ventricular cavity. Epinephrine and dobutamine were stopped, and norepinephrine (noradrenaline) was successfully used to control the hypotension. CONCLUSION: Treatment of hypotension with beta-agonists in VLBW infants with left ventricular hypertrophy can lead to hypercontractility and left ventricular obliteration with paradoxical hypotension. Careful echocardiographic monitoring is indicated in such cases. PMID- 12892165 TI - Characteristics of young foster children in the urban slums of Bangladesh. AB - AIM: To identify the reasons for the fostering of children, and to describe their feeding practices and assess their health and nutritional status. METHODS: Forty six pairs of mothers and their young foster children (up to 24 mo of age) were interviewed in a cross-sectional study in the urban slums of Dhaka, Bangladesh. Another 82 pairs of mothers and their biological children of a similar age group were interviewed for comparison. RESULTS: Sixty-five percent of the children were fostered because of the death of their biological mothers. Eighty-three percent of the biological mothers died because of complications during delivery or the immediate postpartum period and the remainder died after the postpartum period of diseases unrelated to their pregnancies. More than half (52%) of the foster mothers were childless and a quarter (24%) fostered children for reasons of gender preference. Almost a quarter (24%) of the children were placed in foster homes because of extreme parental poverty. Divorce or separation of the biological parents accounted for only 7% of children fostered. Approximately 90% of the foster children were given animal and/or formula milk in their first month of life while all children in the comparison group were given breast milk. Among the foster children, 58% were given semisolid or solid food before completion of 4 mo compared with 14% in the comparison group. Significantly more children in the foster group suffered from diarrhoea and acute respiratory infection than in the comparison group. CONCLUSION: Maternal death, poverty and childlessness and child gender preference of foster mothers were important factors in fostering of children in the study group. Inappropriate feeding practices compounded by diseases may have contributed to the poor nutritional status of the foster children. PMID- 12892166 TI - Factors affecting length of hospitalization of infants and children for recurrent gastroenteritis in Western Australia. AB - AIM: Aboriginal infants in Western Australia (WA) have much higher rates of hospitalization for gastroenteritis than do their non-Aboriginal peers. The aim of this 7-y retrospective study was to investigate the factors affecting the duration of hospitalizations due to recurrent gastroenteritis in WA. METHODS: All infants born in WA in 1995 and 1996 who had indexed gastroenteritis admissions during their first year of life (n = 1009) were followed until 31 May 2002. Linked hospitalization and birth records were retrieved to derive the outcome measures, associated co-morbidities and other demographic variables. A proportional hazards frailty model was then used to determine factors influencing discharge from hospital for the study cohort. RESULTS: Aboriginal patients were readmitted to hospital more frequently and stayed in hospital more than twice as long as non-Aborigines. Factors that slowed the rate of discharge from hospital included Indigenous status and the presence of coexisting conditions such as dehydration, sugar intolerance, failure to thrive and iron-deficiency anaemia. CONCLUSION: Gastrointestinal infections are common illnesses in infants and children, particularly for those who live in overcrowded and unhygienic conditions. Coexisting morbidities including undernutrition, nutrient deficiencies, other infections and gastrointestinal carbohydrate intolerance contribute to prolonged and recurrent hospitalization. Multifaceted clinical, disease prevention and health/hygiene promotion are needed to lessen the burden of these infections. PMID- 12892167 TI - Increasing prevalence of overweight in young schoolchildren in Umea, Sweden, from 1986 to 2001. AB - AIM: To investigate the current prevalence and trend of overweight in young schoolchildren in Umea, Sweden. METHODS: Two cross-sectional samples of children were studied: 1115 randomly selected children from preschool class to grade 6 (aged 6-13 y) attending school in 2001 and a matched sample of 507 schoolchildren from grades 0, 1 and 4 (aged 6-11 y) attending school in 1986. Overweight was defined according to age- and gender-specific body mass index (BMI) cut-off values, corresponding to BMI values of 25 kg/m2 (level 1) and 30 kg/m2 (level 2) at late adolescence. RESULTS: In the population from 2001, the prevalence of overweight was 23% (18% at level 1, 5% at level 2). The prevalence differed with age, with a higher prevalence at the ages of 6 and 13 y. Comparing children from grades 0, 1 and 4, attending school in 1986 and 2001, respectively, the total prevalence of overweight was twice as high in 2001 as in 1986. Regarding severe overweight (level 2), the difference was even larger (five times). In the 1986 sample, the prevalence of overweight did not differ between girls and boys, while significantly more girls than boys were overweight in 2001. CONCLUSION: In this sample of young children from Sweden, the prevalence of overweight doubled over the past 15 y and severe overweight increased even more, suggesting a need for intensified preventive efforts in young schoolchildren. PMID- 12892168 TI - Seven-year review of paediatric bacteraemias diagnosed in a Spanish university hospital. AB - This study analysed the clinical and bacteriological patterns of paediatric bacteraemia in a university hospital, by a review of 213 episodes over a period of 7 y. Streptococcus pneumoniae was the most frequent aetiological agent after the neonatal period and Streptococcus agalactiae in neonatal sepsis. Almost half of pneumococci and meningococci were penicillin non-susceptible. Four neonatal deaths attributed to bacteraemia were recorded. CONCLUSION: Streptococcus pneumoniae is the leading cause of community-acquired bacteraemia. Mortality due to bacteraemia in children without underlying conditions is rare. PMID- 12892169 TI - Pulmonary vein stenosis mimicking chronic lung disease. AB - The presence of recurrent respiratory symptoms and right heart enlargement in an ex-premature infant is suggestive of chronic lung disease. Pulmonary vein stenosis is a rare, progressive disorder that produces similar symptoms and signs. A case is reported in which pulmonary vein stenosis was revealed by Doppler echocardiography, and this application is recommended in similar cases. Pulmonary vein stenosis is an extremely rare and progressive disorder, which, if left untreated, is usually fatal. The possibility of this diagnosis may not be considered during clinical examination and may be overlooked during routine echocardiography. CONCLUSION: This report describes a patient with pulmonary vein stenosis in whom the diagnosis was delayed as she had symptoms and signs more commonly associated with chronic lung disease. PMID- 12892170 TI - Epstein-Barr virus-associated haemophagocytic lymphohistiocytosis in Wiskott Aldrich syndrome. AB - A patient with Wiskott-Aldrich syndrome who developed Epstein-Barr virus associated haemophagocytic lymphohistiocytosis (EBV-HLH) is described in this study. At 4 mo of age the patient developed fever associated with bicytopenia and splenomegaly. Analysis of a bone marrow specimen revealed extensive haemophagocytosis, and in situ hybridization for EBV of the bone marrow specimen using an EBV-encoded RNA probe was positive. Diagnosis of EBV-HLH was established and immunotherapy with HLH-94 protocol was started. HLH has been described in patients with other well-defined primary immunodeficiencies such as X-linked lymphoproliferative syndrome, Chediak-Higashi syndrome and Griscelli disease. Also, HLH was reported recently in severe combined immunodeficiency and DiGeorge syndrome. CONCLUSION: The possibility of an underlying primary immunodeficiency should be considered in paediatric patients who present with HLH during infancy. PMID- 12892171 TI - A concurrent occurrence of cutis laxa, Dandy-Walker syndrome and immunodeficiency in a girl. AB - We report on a 17-y-old girl with inherited cutis laxa, immunodeficiency and Dandy-Walker syndrome. Immunodeficiency manifested itself by decreased and fluctuating levels of IgG, IgA and IgM and intermittent leucopenia causing increased susceptibility to respiratory tract infections. Dandy-Walker syndrome (agenesis of the cerebellar vermis with a large posterior fossa cyst communicating with an enlarged 4th ventricle) was shown on a CT scan but with the exception of macrocrania, no typical signs or symptoms were observed at the age of 17. Loose hyperextensible skin with pendulous skinfolds as a manifestation of cutis laxa was observed from birth. Anomalies of the right pulmonary artery, abnormal branching of the left arteria subclavia (arteria lusoria) from the left aortic arch and bicuspidal aortic valve were also present. CONCLUSION: The combination of the rare disorders cutis laxa, Dandy-Walker syndrome and immunodeficiency is reported here for the first time. PMID- 12892172 TI - Partial trisomy 22 resulting from rearrangements between chromosomes 11/22 and 16/22: a report of two cases. AB - Two male infants with partial trisomy 22 resulting from a rearrangement between chromosomes 11/22 and 16/22 were admitted to the Children's Hospital of the University of Leipzig within the space of two months. The characteristic phenotype of the infants is described and compared with the data on liveborn infants with trisomy 22, as reported in the literature. One of the infants reported here showed a prenatally detected hygroma colli. To the best of our knowledge this is the first description of a hygroma colli in this chromosomal disorder. CONCLUSION: Infants with trisomy 22 can present with variable phenotypes. It is important to bear the phenotype of chromosome 22 infants in mind. PMID- 12892173 TI - Hallermann-Streiff syndrome associated with small cerebellum, endocrinopathy and increased chromosomal breakage. AB - Hallermann-Streiff syndrome (HSS) is a rare clinic entity of unknown aetiology. Further clinical and metabolic-genetic evaluations are indicated. A 2-mo-old female baby presented with ocular abnormalities and severe failure to thrive since birth. The clinical features were compatible with the diagnosis of HSS. Further imaging, metabolic and cytogenetic examinations were performed. Features characteristic of HSS were dyscephaly with mandibular and nasal cartilage hypoplasia, microphthalmia, bilateral cataracts with congenital glaucoma, natal teeth and proportionate dwarfism. Rare anomalies such as choanal atresia and small cerebellum, very low insulin-like growth factor I level, hypothyroidism, generalized organic aciduria were also noticed. An increased chromosomal breakage rate is suggestive of the existence of some DNA repair defects in HSS patients. CONCLUSION: The associated anomalies in this patient may broaden the clinical spectrum of HSS. Underlying conditions of organic aciduria, growth factor deficiency and impaired DNA repair are likely to contribute to the progeria-like facies, congenital cataracts and growth failure. PMID- 12892174 TI - Summary of confirmed cases of atypical pneumonia (SARS) in Hong Kong: daily update. PMID- 12892175 TI - Predictors of outcome. Comment on the article by Eriksson et al. PMID- 12892176 TI - No evidence for complete catch-up in final height in psychosocial short stature. PMID- 12892177 TI - Intra-pelvic sacrococcygeal teratoma. PMID- 12892178 TI - Trisomy 5 in acute lymphoblastic leukaemia. PMID- 12892179 TI - Diagnosing acute compartment syndrome. PMID- 12892180 TI - The physiology of the disc and its clinical relevance. PMID- 12892181 TI - Can we improve the results of revision arthroplasty for infected total hip replacement? PMID- 12892182 TI - Cementless total hip arthroplasty in patients older than 80 years of age. AB - We reviewed 80 patients (87 hips) who were older than 80 years of age at the time of cementless total hip arthroplasty. An Alloclassic SL stem had been implanted in all patients. A variety of cementless acetabular components was used. After a mean follow-up of 69.3 months (39.2 to 94.1) 48 hips in 43 patients were analysed clinically and radiologically. One patient had sustained a traumatic periprosthetic fracture of the femur with subsequent exchange of the stem 73 months after operation. Thirty-two patients (34 hips) had died and five patients (five hips) were unavailable for follow-up because of health reasons (four patients) or lack of co-operation (one patient). If the endpoint is defined as removal of the prosthesis because of aseptic loosening, the survival rate was 100% for the cup and stem after 78 months. The mean Harris hip score was 81.9 points. Radiolucent lines and osteolysis were seldom found. PMID- 12892183 TI - The pharmacokinetics of Simplex-tobramycin bone cement. AB - We prospectively investigated a consecutive series of ten patients undergoing a cemented primary total hip replacement (THR) for osteoarthritis in order to establish the elution characteristics of Simplex-tobramycin bone cement (Howmedica, Limerick, Ireland). Specimens of blood, urine and drainage fluid were collected for 72 hours postoperatively. Very high concentrations of tobramycin were found in the drainage fluid, with mean levels at one hour of 103 mg/l, which steadily declined to 15.1 mg/l after 48 hours. The mean serum tobramycin levels reached a peak of 0.94 mg/l at three hours and declined rapidly to 0.2 mg/l by 48 hours. The mean urinary tobramycin levels peaked at 57.8 mg/l at 12 hours with a rapid decline to 12.6 mg/l by 24 hours. There was a direct correlation between the amount of tobramycin bone cement which was implanted and the amount of tobramycin systemically absorbed. Excellent local delivery was achieved with minimal systemic concentrations. Simplex-tobramycin bone cement is an efficient and safe method for the delivery of antibiotics after THR. PMID- 12892184 TI - Dislocation after total hip replacement in relation to metal-on-metal bearing surfaces. AB - Metal-on-metal (MOM) is a commonly used bearing notable for its 'suction fit' when lubricated. In this study, we examined the capacity for MOM bearings to protect against dislocation after total hip replacement (THR). We undertook a clinical investigation to compare the rate of dislocation of MOM bearings with those of ceramic-on-polyethylene (COP) bearings and found that one MOM bearing dislocated in a series of 109 hips (0.9%) compared with nine of 145 hips (6.2%) in the COP group (p = 0.02). We also performed an in vitro investigation comparing the peak forces generated during forced separation of the two bearings of the same dimensions at velocities from 1 to 50 cm/s. This revealed that the MOM bearing generated significant resistance to separation at all velocities (maximum mean 24 N), whereas the COP did not (maximum mean 1.9 N, p < 0.001). We conclude that MOM bearings are more stable to dislocation than COP bearings as a result of the interfacial forces provided by a thin, lubricating fluid. PMID- 12892185 TI - Effect of alumina femoral heads on polyethylene wear in cemented total hip arthroplasty. Old versus current alumina. AB - We examined the behaviour of alumina ceramic heads in 156 cemented total hip arthroplasties, at a minimum follow-up of eight years. They were divided into three groups according to the size of the femoral head; 22, 26, and 28 mm. We measured polyethylene wear radiologically using a computer-aided technique. The linear wear rate of polyethylene sockets for the 28 mm heads was high (0.156 mm/year), whereas those for the 22 and 26 mm heads were relatively low (0.090 and 0.098 mm/year, respectively). Moreover, the surface roughness data of retrieved femoral heads clearly showed maintenance of an excellent surface finish of the current alumina. We conclude that the alumina ceramic femoral heads currently used are associated with a reduced rate of polyethylene wear. PMID- 12892186 TI - Incidence and natural history of deep-vein thrombosis after total hip arthroplasty. A prospective and randomised clinical study. AB - There are many reports concerning the aetiology and prophylaxis of deep-vein thrombosis (DVT) but little is known about its natural history. The purpose of our study was to identify the incidence and site of DVT, the risk factors for pulmonary embolism and the natural history of DVT after total hip replacement (THR) in patients who do not receive any form of prophylactic or therapeutic treatment for DVT. Two hundred patients who had a primary THR were included: 100 had one-staged bilateral THR and 100 had unilateral THR and 150 implants were cemented and 150 cementless. Coagulation assays, a full blood count, blood typing and serum chemical profile tests were performed for all patients on three separate occasions. Bilateral simultaneous or unilateral venograms were performed on the sixth or seventh postoperative day and perfusion lung scans preoperatively and on the seventh or eighth postoperative day. Further venograms were performed in all patients who had thrombi six months later. In the patients with bilateral THR, 52 (26%) venograms were positive for thrombi, while in the patients with unilateral THR 20 (20%) were positive (p = 0.89). In the patients with a cemented THR, 31 venograms (20.7%) were positive for thrombi, while in those with a cementless THR 41 (27.3%) were positive (p = 0.654). Further venograms in all 72 patients who had thrombi at six months after operation showed that they resolved completely and spontaneously regardless of their site and size. No patients had symptoms of pulmonary emboli and none were seen on the perfusion lung scans. Two patients died from unrelated causes. Although the prevailing opinion is that patients with proximal venous thrombosis should be treated with anticoagulants, our study has shown that all thrombi regardless of their site and size resolve spontaneously without associated pulmonary embolism. PMID- 12892187 TI - Arthrodesis of the shoulder after septic arthritis. Long-term results. AB - We analysed the long-term results of arthrodesis of the shoulder after infection in 15 patients. At the time of operation, 14 cultures were positive for Staphylococcus aureus. The mean follow-up was 8.3 years (3 to 14) and 90% of the patients were satisfied with the outcome. There were complications in five patients (33%); in three there was nonunion with loosening of the implant. One patient had a sound bony union but with a persistent sinus six years after arthrodesis and another had a sinus which healed after the metal was removed. Four of these five patients (80%) were heavy smokers (> 20 cigarettes/day). Cancellous bone grafting did not affect the incidence of complications. The mean age of the patients with complications was 58.6 v 48.6 years for those without (p = 0.2808; not significant). Those with complications had had more previous operations (6.4 v 2.5, p < 0.05). Antibiotics, as determined by the bacteriological cultures, were administered for six weeks. The complication rate was higher in patients with active sepsis but the younger the patient and the fewer number of previous operations (< 50 years, < four previous operations), the better was the outcome. Considering the rate of complications, we recommend early surgery in these patients. PMID- 12892188 TI - Posterior cruciate ligament at total knee replacement. Essential, beneficial or a hindrance? AB - We report the results of a prospective randomised trial which assessed the role of the posterior cruciate ligament (PCL) following total knee replacement (Genesis I; Smith and Nephew, Memphis, Tennessee). Over a four-year period, 211 patients underwent total knee replacement by the senior author (TJW). They were randomised at surgery to have the PCL either retained, excised or substituted with a posterior stabilised insert. If it was not possible to retain the ligament due to soft-tissue imbalance, it was released from its tibial insertion until suitable tension was obtained. This created a fourth group, those who were intended preoperatively to have the ligament retained, but in whom it was partially released as a result of findings at the time of surgery. All patients were evaluated using the Knee Society rating system (adapted from Insall). A total of 188 patients (212 knees) was available for follow-up at a mean of 3.5 years after surgery. Preoperatively, there was a varus deformity in 191 knees (90%) and a valgus deformity in 21 (10%). There were no statistical differences in the knee or function scores or the range of movement between the excised, retained and substituted groups. There were, however, significantly worse knee and function scores in the group in whom the PCL was released (p = 0.002). PMID- 12892189 TI - Secondary resurfacing of the patella for persistent anterior knee pain after primary knee arthroplasty. AB - Out of a total of 623 patients who, over a ten-year period, underwent primary total knee replacement (TKR) without patellar resurfacing, 20 underwent secondary resurfacing for chronic anterior knee pain. They were evaluated pre- and postoperatively using the clinical and radiological American Knee Society score. The mean follow-up was 36.1 months (12 to 104). The mean knee score improved from 46.7 to 62.2 points and the mean functional score from 44.7 to 52.2 points. Only 44.4% of the patients, however, reported some improvement; the remainder reported no change or deterioration. The radiographic alignment of the TKR did not influence the outcome of secondary resurfacing of the patella. Complications were noted in six of the 20 patients including fracture and instability of the patella and loss of movement. Anterior knee pain after TKR remains difficult to manage. Secondary resurfacing of the patella is not advocated in all patients since it may increase patient dissatisfaction and hasten revision. PMID- 12892190 TI - Oxidative stress in bilateral total knee replacement, under ischaemic tourniquet. AB - Free radicals, such as reactive oxygen species (ROS) which are released abruptly after deflation of an ischaemic tourniquet, cause reperfusion injuries. Ischaemic precondition (IPC), however, can reduce the injury. In clinical practice, the sequential application and release of tourniquets is often used in bilateral total knee replacement (TKR) to obtain a clearer operative field, but the effects on the production of free radicals and lipid peroxidation have not been studied. In this study, we have observed the production of free radicals and the subsequent lipid peroxidation in bilateral TKR with sequential application of a tourniquet to examine the effect of IPC. Patients undergoing elective TKR under intrathecal anaesthesia were studied. Blood samples were obtained after spinal anaesthesia, one minute before and five and 20 minutes after release of each tourniquet. We used the lucigenin chemiluminescence analysis and the phosphatidylcholine hydroperoxide (PCOOH) assay to measure the production of ROS and lipid peroxidation. Our results showed that production of ROS significantly increased at five and 20 minutes after release of the first tourniquet and at five minutes after release of the second tourniquet, but returned to normal at 20 minutes after the second reperfusion. The peak production of ROS was at 20 minutes after the first reperfusion; lipid peroxidation did not change significantly. We conclude that in spite of significant production of ROS after the release of tourniquet, the IPC phenomenon occurs during bilateral TKR with sequential application of a tourniquet. PMID- 12892191 TI - The management of brachymetatarsia. AB - We treated 35 brachymetatarsal rays of 18 feet in 12 patients by one-stage lengthening with interpositional bone grafts or by gradual lengthening with callotasis combined with shortening of the adjacent metatarsals and phalanges. Definition of the two parabolas which connect the metatarsal heads and the tips of the toes, and recognition of three patterns of metatarsal length, were helpful guides in treatment. In total, 36 excisions of the phalanges and/or the metatarsals were undertaken. The mean shortening was 8 mm. The radiological results were satisfactory. The mean values were as follows: one-stage lengthening, length gain, 1.3 cm; healing index, 1.3 months/cm; percentage increase, 30%; gradual lengthening, length gain, 2.0 cm; healing index, 2.0 months/cm; percentage increase, 50%. Associated shortening of an adjacent bone can avoid the disadvantages of one-stage lengthening which may not achieve target length and can result in neurovascular complications. Reduction of the target length enables the surgeon to carry out one-stage instead of gradual lengthening. It also shortens the length of treatment in the group undergoing callotasis and improves cosmesis. PMID- 12892192 TI - Outcome of complex fractures of the tibial plateau treated with a beam-loading ring fixation system. AB - Fine-wire external fixation is accepted as a minimally invasive technique, which can provide better outcomes than traditional open methods in the management of complex fractures of the tibial plateau. Available fixators vary in their biomechanical stability, and we believe that a stable beam-loading system is essential for consistently good outcomes. We assessed, prospectively, the clinical, radiological and general health status of 20 of 21 consecutive patients with complex fractures of the tibial plateau who had been treated using a standard protocol, with percutaneous screw fixation and a neutralisation concept with a fine wire beam-loading fixator allowing early weight-bearing. Bony union was achieved in all patients, with 85% having good or excellent results. Full weight-bearing started during the first six weeks in 60% of patients. The general health status assessment correlated well with the knee scores and reflected a satisfactory outcome. PMID- 12892193 TI - Heterotopic ossification prophylaxis with indomethacin increases the risk of long bone nonunion. AB - Indomethacin is commonly administered for the prophylaxis of heterotopic ossification (HO) after the surgical treatment of acetabular fractures. Non steroidal anti-inflammatory drugs such as indomethacin, have been associated with delayed healing of fractures and mechanically weaker callus. Our aim was to determine if patients with an acetabular fracture, who received indomethacin for prophylaxis against HO, were at risk of delayed healing or nonunion of any associated fractures of long bones. We reviewed 282 patients who had had open reduction and internal fixation of an acetabular fracture. Patients at risk of HO were randomised to receive either radiation therapy (XRT) or indomethacin. Of these patients, 112 had sustained at least one concomitant fracture of a long bone; 36 needed no prophylaxis, 38 received focal radiation and 38 received indomethacin. Fifteen patients developed 16 nonunions. When comparing patients who received indomethacin with those who did not, a significant difference was noted in the rate of nonunion (26% v 7%; p = 0.004). Patients with concurrent fractures of the acetabulum and long bones who receive indomethacin have a significantly greater risk of nonunion of the fractures of the long bones when compared with those who receive XRT or no prophylaxis. PMID- 12892194 TI - Development and testing of a reduced WOMAC function scale. AB - We used prospective data from 862 total knee and 716 total hip replacements three years after surgery in order to derive and validate a reduced Western Ontario and McMasters University Osteoarthritis Index (WOMAC) function scale. The reduced scale was derived using the advice of clinical experts as well as analysis of data. The scale was tested for validity, reliability and responsiveness. Items which were retained included: ascending stairs, rising from sitting, walking on the flat, getting in or out of a car, putting on socks, rising from bed, and sitting. The reduced and full scales had comparable, moderate correlations with other measures of function, confirming convergent validity. Cronbach's alpha was high (alpha > 0.85) with the reduced scale confirming reliability. Responsiveness was greater for the reduced scale (full = 1.4, reduced = 1.6). This reduced version of the WOMAC function scale provides a practical, valid, reliable and responsive alternative to the full function scale for use after total joint replacement. Further work is needed to demonstrate its wider applicability. PMID- 12892195 TI - Acrylic cement added with antiblastics in the treatment of bone metastases. Ultrastructural and in vitro analysis. AB - An increased long-term survival of patients with alignant tumours also increases the possibility of the development of skeletal metastases and pathological fractures. The management of bone metastases includes the removal of gross disease and the administration of local adjuvants. We have investigated the possibility of adding antiblastic drugs to acrylic cement. Cylinders of acrylic cement were manufactured containing three different antiblastic drugs, methotrexate, cisplatin and doxorubicin. We performed in vitro analysis on MCF-7 human breast cancer cells in order to evaluate the biological effect of the mixtures and surface analysis of the acrylic cement-cisplatin cylinders using energy-dispersive x-ray analysis (EDAX). All drugs were released in an active form from the cement. Each drug had a different effect on cell viability. Doxorubicin had the greatest effect on breast cancer cells. Surface analysis showed that antiblastic drugs were present in the form of granules. These results confirm the potential of antiblastic-loaded cement as a possible adjuvant in the local treatment of bone metastases. Further studies should be undertaken to determine whether the release of antiblastic drugs from cement is elution or if they are only released from the surface. PMID- 12892196 TI - Endoprosthetic replacement of the proximal humerus. Long-term results. AB - We studied 100 patients who had undergone endoprosthetic replacement of the proximal humerus between 1976 and 1998. The outcome was assessed with regard to the survivorship of the patients, the salvaged limbs and the prostheses. Function was determined in the 47 surviving patients, of whom 30 were assessed using the Musculo-Skeletal Tumour Society (MSTS) rating scale and 38 completed the Toronto Extremity Salvage Score (TESS) questionnaire. The median age of the patients was 34 years (10 to 80). The mean follow-up period for surviving patients was nine years (2 to 20). The mean MSTS score at follow-up was 79% and the mean TESS score was 72%. The length of bone which was resected influenced the functional outcome. Abduction of the shoulder was to 45 degrees in most patients. The overall survival was 42% at ten years and that of the limb without amputation 93%. The survival of the prostheses using mechanical failure as the endpoint was 86.5% at 20 years. Endoprosthetic replacement of the proximal humerus is a predictable procedure providing reasonable function of the arm with a low rate of complications at long-term follow-up. PMID- 12892197 TI - Intramuscular neurilemoma. AB - In this retrospective study, we analysed the clinical features of neurilemoma when it is located in muscle. Twelve patients had an intramuscular neurilemoma as shown on magnetic resonance (MR) scans and confirmed at operation. In six it was located in the upper limb, in five in the lower limb, and in one in the back. The mean age of the patients was 41 years (12 to 58). Nine complained only of a palpable mass and the other three of a mass with slight tenderness. None had neurological symptoms or signs, such as radicular pain, a positive Tinel sign, or motor weakness. There were no postoperative complications or recurrence of the tumour after a mean follow-up of two years (1 to 10). PMID- 12892198 TI - Ultrasonographic screening for developmental dysplasia of the hip in infants. Reproducibility of assessments made by radiographers. AB - We studied the reproducibility of ultrasonographic screening examination of the hip when read by diagnostic radiographers. In order to determine interobserver variability, 200 ultrasonograms were classified according to Graf's method by five observers (four radiographers and one radiologist). The kappa values for interobserver variability indicated moderate agreement (kappa 0.47) for the exact Graf classification and substantial agreement (kappa 0.65) for the classification of normal (type I) versus abnormal (type IIa-IV). Agreement was significantly different for normal, immature and abnormal hips. Comparison of the findings in our interobserver study with existing information based on other examinations and treatment revealed that only a small number of infants with mildly dysplastic hips would have been typed as normal by some observers as a result of observer variability. In conclusion, the interobserver agreement on the ultrasound assessment of the hip was good enough for screening purposes. Observer variability did not result in any severe cases being missed. PMID- 12892199 TI - Instability after total knee replacement with a mobile-bearing prosthesis in a patient with multiple sclerosis. AB - We describe a patient with multiple sclerosis (MS), who developed recurrent dislocations after total knee arthroplasty. She had both knees replaced using similar mobile-bearing knee prostheses, but the outcome was worse in the leg which was more severely affected with MS. PMID- 12892200 TI - Histoplasmosis as the cause of a pathological fracture. AB - We report the case of an 82-year-old man with a pathological fracture of the hip caused by infection with Histoplasma capsulatum var capsulatum. He was treated by a hemiarthroplasty and with oral itraconazole. PMID- 12892201 TI - Candida septic arthritis of the hip in a young patient without predisposing factors. AB - We present a case of septic arthritis of the hip caused by Candida albicans in a 24-year-old woman. This is the first report of a Candida infection of the hip in a patient without any predisposing factors. She underwent a two-stage total hip arthroplasty three years after the onset of the infection. An elevated plasma level of beta-D-glucan suggested Candida as the infecting organism and the diagnosis might have been made earlier if this test had been undertaken at presentation. PMID- 12892202 TI - Pencil-core granuloma of the distal radio-ulnar joint: an unusual presentation as soft-tissue sarcoma after 45 years. AB - A case of pencil-gore granuloma occurring 45 years after he initial injury is described. The clinical and radiological presentation was suggestive of a soft tissue sarcoma. The diagnosis was confirmed by biopsy, which also revealed foreign body fragments which, on spectometric analysis, had high concentrations of carbon and aluminium. PMID- 12892203 TI - Tissue-engineered cartilage and bone using stem cells from human infrapatellar fat pads. AB - Multipotential processed lipoaspirate (PLA) cells extracted from five human infrapatellar fat pads and embedded into fibrin glue nodules, were induced into the chondrogenic phenotype using chondrogenic media. The remaining cells were placed in osteogenic media and were transfected with an adenovirus carrying the cDNA for bone morphogenetic protein-2 (BMP-2). We evaluated the tissue-engineered cartilage and bone using in vitro techniques and by placing cells into the hind legs of five severe combined immunodeficient mice. After six weeks, radiological and histological analysis indicated that the PLA cells induced into the chondrogenic phenotype had the histological appearance of hyaline cartilage. Cells transfected with the BMP-2 gene media produced abundant bone, which was beginning to establish a marrow cavity. Tissue-engineered cartilage and bone from infrapatellar fat pads may prove to be useful for the treatment of osteochondral defects. PMID- 12892204 TI - Influence of peg design and cement mantle thickness on pull-out strength of glenoid component pegs. AB - Fixation of the glenoid component is critical to the outcome of total shoulder arthroplasty. In an in vitro study, we analysed the effect of surface design and thickness of the cement mantle on the pull-out strength of the polyethylene pegs which are considered essential for fixation of cemented glenoid components. The macrostructure and surface of the pegs and the thickness of the cement mantle were studied in human glenoid bone. The lowest pull-out forces, 20 +/- 5 N, were for cylindrical pegs with a smooth surface fixed in the glenoid with a thin cement mantle. The highest values, 425 +/- 7 N, were for threaded pegs fixed with a thicker cement mantle. Increasing the diameter of the hole into which the peg is inserted from 5.2 to 6.2 mm thereby increasing the thickness of the cement mantle, improved the mean pull-out force for the pegs tested. PMID- 12892205 TI - The origin of cells that repopulate patellar tendons used for reconstructing anterior cruciate ligaments in man. AB - Cryopreserved patellar tendon allografts are often recommended for reconstruction of anterior cruciate ligaments (ACLs) because living donor fibroblasts are thought to promote repair. Animal studies, however, indicate that ligaments regenerate from recipient rather than donor cells. If applicable to man, these observations suggest that allograft cell viability is unimportant. We therefore used short tandem repeat analysis with polymerase chain reaction (PCR) amplification to determine the source of cells in nine human ACLs reconstructed with cryopreserved patellar tendon allografts. PCR amplification of donor and recipient DNA obtained before operation and DNA from the graft obtained two to ten months after transplantation revealed the genotype of cells and showed only recipient cells in the graft area. Rather than preserve the viability of donor cells, a technique is required which will facilitate the introduction of recipient cells into patellar tendon allografts. PMID- 12892206 TI - Immunological changes in patients with primary osteoarthritis of the hip after total joint replacement. AB - We aimed to assess whether the immunological abnormalities which have been observed in patients with loose total hip replacements (THRs) are present in patients with a well-fixed prosthesis. We examined blood samples from 39 healthy donors, 22 patients before THR and 41 with well-fixed THRs of different types (15 metal-on-metal, 13 metal-on-polyethylene, 13 ceramic-on-ceramic). Before THR, the patients showed a decrease in leukocytes and myeloid cells in comparison with healthy donors, and a prevalence of type-1 T lymphocytes, which was confirmed by the increase in ratio of interferon-gamma to interleukin 4. Moreover, patients with metal-on-metal or metal-on-polyethylene implants showed a significant decrease in the number of T lymphocytes and a significant increase in the serum level of chromium and cobalt, although no significant correlation was observed with the immunological changes. In the ceramic-on-ceramic group, leukocytes and lymphocyte subsets were not significantly changed, but a significant increase in type-2 cytokines restored the ratio of interferon-gamma to interleukin 4 to normal values. We conclude that abnormalities of the cell-mediated immune response may be present in patients with a well-fixed THR, and that the immunological changes are more evident in those who have at least one metal component in the articular coupling. PMID- 12892207 TI - The meniscofemoral ligaments: secondary restraints to the posterior drawer. Analysis of anteroposterior and rotary laxity in the intact and posterior cruciate-deficient knee. AB - We have tested the hypothesis that the meniscofemoral ligaments make a significant contribution to resisting anteroposterior and rotatory laxity of the posterior-cruciate-ligament-deficient knee. Eight cadaver human knees were tested for anteroposterior and rotatory laxity in a materials-testing machine. The posterior cruciate ligament (PCL) was then divided, followed by division of the meniscofemoral ligaments (MFLs). Laxity results were obtained for intact, PCL deficient, and PCL-MFL-deficient knees. Division of the MFLs in the PCL-deficient knee increased posterior laxity between 15 degrees and 90 degrees of flexion. Force-displacement measurements showed that the MFLs contributed 28% to the total force resisting posterior drawer at 90 degrees of flexion in the intact knee, and 70.1% in the PCL-deficient knee. There was no effect on rotatory laxity. This is the first study which shows a function for the MFLs as secondary restraints to posterior tibial translation. The integrity of these structures should be assessed during both imaging and arthroscopic studies of PCL-injured knees since this may affect the diagnosis and management of such injuries. PMID- 12892208 TI - Extracorporeal shock-wave therapy for tendonitis of the rotator cuff. PMID- 12892209 TI - Arthroplasty of the hip--leg length is not important. PMID- 12892210 TI - The role of angiography in the management of haemorrhage from major fractures of the pelvis. PMID- 12892211 TI - The type of cement and failure of total hip replacements. PMID- 12892212 TI - Intraneural mucoid pseudocysts: a report of ten cases. PMID- 12892213 TI - Development of remote control and monitoring system for the FE gun in HVEM. AB - This paper describes a control and monitoring system for the field emission (FE) gun in the high-voltage electron microscope (HVEM), where the whole FE gun system is located inside a pressure vessel and is floated on a high voltage of 1 MV. The operating condition of the FE gun system is controlled and monitored mainly by a bi-directional communication system between ground and high potential through optical fibres. A mechanical driving system by combination of insulator rods and stepping motors is partially used as a control system for FE gun operation. The monitoring system developed here also functions as a safety circuit that detects abnormal situations, such as an abnormal deterioration of vacuum, and protects the FE tip against fatal damage. The fundamental performance of the microscope has been tested for transmission electron microscope (TEM) and scanning TEM operation modes. PMID- 12892214 TI - Characterization of polymer-adhesive interfaces on a nanometre scale by elemental mapping and image EELS using EFTEM. AB - Energy-filtering transmission electron microscopy (EFTEM) was applied for investigating interfaces between a polymer and an adhesive. The sample employed in this work is polybutylene terephtharate (PBT) sheets laminated with an epoxy adhesive. It was found that heat aging of the PBT at 180 degrees C in air for > 9 h prior to adhesion decreases the adhesion strength drastically. To investigate this unfavourable aging effect on the adhesion strength, we performed elemental mapping and image EELS using EFTEM. A weak boundary layer with a thickness of < 50 nm was visualized at the PBT-adhesive interface by elemental mapping in the sample subjected to the heat aging and image EELS revealed the origin of this layer. Thus, we clearly correlated the nanoscale interfacial structure with the adhesion strength by EFTEM. PMID- 12892215 TI - Connection structures between type-I and type-II defects in neutron irradiated beta-Si3N4. AB - High-resolution electron microscopy (HREM) observation clarified that various types of interstitial dislocations were induced into beta-Si3N4 by high-dose neutron irradiation, and two of them have been analysed and reported in our previous papers. These two defective structures are called type-I and type-II. They are interstitial dislocation loops introduced into [100] and [110] planes, respectively. In this study, it was found that some of these dislocation loops connected with each other. The connections of type-I-L2-type-II-B and type-I-R2 type-II-A dislocations were observed by HREM. Tetrahedral arrangements of the connected parts are proposed, based on the simplified crystal structure model of beta-Si3N4. In addition, a new type of defect, type-III, was found. Type-III is an interstitial dislocation loop introduced into [100] planes. It was also found that type-I-A and type-III dislocations connected with each other. PMID- 12892216 TI - Microdomain structure of agar gels observed by mechanical-scanning probe microscopy. AB - The inhomogeneous structure of agar gels was examined by means of mechanical scanning probe microscopy. Several domains were observed in the elasticity images, while such domains could not be seen in the height images. The domain size decreased with increases in agar concentration. We found that the histograms of the logarithm of the local elastic modulus were described well by a single normal distribution. As the agar concentration increased, the peak values of the histograms increased, while the half-value width remained constant. These results imply that the gelation process of agar gels has a common mechanism, despite its complexity. PMID- 12892217 TI - Elemental mapping of DNA chains by energy-filtering TEM. AB - DNA chains were detected by phosphorus mapping based on energy-filtering transmission electron microscopy (EF-TEM). The three-window method and the image merging system were used to make phosphorus-mapping images. It was impossible to observe DNA chains without the image-merging system, because of the low signal levels of single images. It became easy to assign DNA molecules when the typical superstructure of plasmids was preserved. To preserve the structure during specimen preparation, rapid-freezing and freeze-drying methods were used. Mapping images can be obtained only when carbon-supporting films are extremely thin, as thick supporting films weaken the signals. The thickness of the supporting film, which was estimated to be < 2 nm, was the most important factor in this study. In the three-window method, the calculation to remove the background absolutely includes a few errors, because the precise spectrum was not observed. To obtain higher quality mapping images, several improvements in the hardware are suggested. PMID- 12892218 TI - Comparative study of depth and lateral distributions of electron excitation between scanning ion and scanning electron microscopes. AB - In order to study the contrast difference between scanning ion microscopes (SIM) and scanning electron microscopes (SEM), the depth and lateral distributions of secondary electrons escaped from surfaces of 17 metals with atomic numbers, Z2, of 4-79 were calculated for bombardment with 30 keV Ga ions and for 10 keV electrons. For both projectiles, the excitation depth generally decreased with increasing Z2, while showing the same periodic change as the secondary-electron yield. However, an opposite trend in Z2 dependence between the Ga ion and electron bombardments was calculated with the lateral distribution of secondary electrons escaped from the surface. Except for low Z2 metals, the lateral distribution, which is much narrower for 30 keV Ga ions than for 10 keV electrons, indicates that the spatial resolution of the secondary-electron images is better for SIM than for SEM, if zero-sized probe beams are assumed. Furthermore, the present calculation reveals important effects of electron excitation by recoiled material atoms and reflected electrons on the lateral distribution, as well as the secondary-electron yield, for the Ga ion and electron bombardments, respectively. PMID- 12892219 TI - The study of Al-L23 ELNES with resolution-enhancement software and first principles calculation. AB - Resolution-enhancement software and first-principles calculation for electron energy-loss spectroscopy (EELS) were applied for the study of aluminium (Al) coordination in alumina. Al-L23 energy-loss near-edge structures (ELNES) of alpha and gamma-aluminas were observed using a field-emission transmission electron microscope and advanced software for EELS. The inherent energy resolution of a cold field-emission gun (FEG) of approximately 0.3 eV was realized using drift correction software. The energy spread of the cold FEG was deconvoluted by means of maximum-entropy or Richardson-Lucy algorithms and the energy resolution of the deconvoluted spectrum became comparable with that obtained using a monochromator, whose energy resolution was < 0.2 eV. Fine structures in Al L23-edge were observed, such as 0.5 eV splitting between L3 and L2 peaks in alpha-alumina (i.e. spin-orbit splitting). The difference in Al coordination was clearly observed in Al-L23 ELNES and the major structures near the threshold were assigned using first-principles calculations. PMID- 12892220 TI - Analytical electron microscopy study of nanometre-scale oxide formed in contact hole-bottom Si surfaces. AB - Nanometre-scale interfacial oxides formed in contact-hole-bottom Si surfaces were investigated by analytical electron microscopy coupled with a field-emission transmission electron microscope. The results showed that the chemical state of the residual oxide formed during reactive-ion etching was mostly changed from the suboxide of Si2+ or Si3+ to the oxide of Si4+ by the following light-etch treatment. Consequently, the process of removing the residual oxide by light-etch treatment was improved and it contributed to the accomplishment of lower contact resistance. PMID- 12892221 TI - Microstructural investigation of ceria-zirconia solid solution with oxygen vacancies. AB - The microstructural change in CeO2-ZrO2 solid solution (Ce2Zr2O7 + x; 0 < or = x < or = 1) with an ordered arrangement of Ce and Zr associated with the absorption/release of oxygen was observed by transmission electron microscopy (TEM). The oxidation of Ce2Zr2O7 was observed in TEM by electron diffraction analysis. The change in the electron diffraction pattern shows that Ce2Zr2O7 was easily oxidized in the TEM and that the oxidation was completed, giving an intermediate phase Ce2Zr2O7.5. Three different lattice images corresponding to x = 0, 0.5 and 1.0 were observed in high-resolution photographs of Ce2Zr2O7 + x. This difference in contrast image may be used for determining the local amount of oxygen absorbed. PMID- 12892222 TI - Ultrastructural basis for the transition of cell death mode from apoptosis to necrosis in menadione-treated osteosarcoma 143B cells. AB - Time-dependent ultrastructural changes of menadione-treated human osteosarcoma 143B cells were correlated with those in their stainability to Annexin V and propidium iodide (PI). Populations of both apoptotic (Annexin V(+)/PI(-)) and necrotic (Annexin V(+)/PI(+)) cells, judged by flow cytometry, began to increase at 2 h after menadione treatment. The former reached a maximum at 6 h followed by abrupt decreases thereafter, while the latter continued to increase. Electron microscopically, cells obtained at 6 h after the menadione treatment consisted of mixed populations of cells with typical apoptotic features and those with a mixture of apoptotic and necrotic features, while cells obtained at 8-24 h consisted exclusively of cells with a mixture of apoptotic and necrotic features. Thus, necrotic cells, as judged by flow cytometry, were in a transitional state of cell death mode from apoptosis to necrosis and are thus designated as 'intermediate cells'. Lack of apoptotic bodies, judged by flow cytometric analysis on sub-G1 nuclei and by electron microscopy in menadione-treated cells, suggested that the transition of cell death mode from apoptosis to necrosis occurred before the apoptotic processes were completed. Effects of N acetylcysteine and Z-VAD-fmk on menadione-induced ultrastructural changes were also studied. PMID- 12892223 TI - A novel morphological technique to investigate a single climbing fibre synaptogenesis with a Purkinje cell in the developing mouse cerebellum: DiI injection into the inferior cerebellar peduncle. AB - To study morphologically the relationship between climbing fibre and Purkinje cell in the developing mouse cerebellum, we established a novel tract tracing using injection of 1,1'-dioctodecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiI) into the inferior cerebellar peduncle, the half point of olivocerebellar projection. In this tracing method, only a certain number of climbing fibres were labelled with DiI and they revealed the single-fibre resolution, individually. These technical advantages enabled us to follow the projection of a climbing fibre to a Purkinje cell at the light microscopic level. To further investigate how a single labelled olivocerebellar axon interacts with a Purkinje cell, we introduced a photoconversion method into this tracing method and successfully observed the photo-oxidized climbing fibre terminals at the electron microscopic level. At postnatal days 7 and 9, a single DiI-labelled climbing fibre arborized around some adjacent Purkinje cell bodies in a distinguishable nest. At this pericellular nest stage, we first demonstrated that the terminal arborization stemmed from a single climbing fibre formed synapses simultaneously on both a soma and dendrites of a Purkinje cell. This finding suggests that the pericellular nest may be such an efficient form that a single climbing fibre innervates a Purkinje cell at both perisomatic and peridendritic sites. Thus, we succeeded in establishing an effective tracing method to investigate a single climbing fibre synaptogenesis with a Purkinje cell both at the light and electron microscopic levels. PMID- 12892224 TI - Ultrastructural organization of the transverse tubules and the sarcoplasmic reticulum in a fish sound-producing muscle. AB - The ultrastructural basis for the extremely rapid contraction-relaxation cycle (up to 300 s(-1)) in the swim-bladder muscle (SBM) of a scorpionfish (Sebastiscus marmoratus), producing characteristic sounds for communication, was investigated by electron microscopy. The SBM fibres contained well-developed sarcoplasmic reticulum (SR) showing triadic contacts with well-organized transverse tubules (T tubules). It was newly found that different types of triadic contacts were present within the single SBM fibre. In the middle region of the fibre (approximately 54% of the fibre length), the triadic contacts were located around the level of boundary between the A- and I-bands (AI-type triad). However in the two end regions of the fibre (approximately 21% and approximately 12% of the fibre length), the triadic contacts were seen around the level of the Z-band (Z type triad). Between the middle and end regions of the fibre, T tubule-SR contacts exhibited the form of pentads composed of a pair of T tubules and three SR elements, and newly found heptads composed of three T tubules and four SR elements. The fractional volume of SR relative to the fibre volume was estimated to be approximately 26% in the middle region of the fibre with the AI-type triads and approximately 15% in the fibre ends with the Z-type triads. These results are discussed in connection with the mechanism, by which the mechanical activity of the SBM muscle is neurally controlled. PMID- 12892225 TI - Elements and organic substances in epiretinal proliferative tissue excised during vitreous surgery: analysis by time-of-flight secondary-ion mass spectrometry. AB - We examined with time-of-flight secondary-ion mass spectrometry (TOF-SIMS) various substances in vitreous proliferative tissues excised from patients with proliferative diabetic retinopathy or proliferative vitreoretinopathy. Amounts of elements, vitamins and fatty acids were measured with TOF-SIMS and then the average ratio of the count of spattered ions of each element and organic substance to the total count of spattered ions was calculated. The average ratios of Fe, Ca, Al, Zn and Cu ion counts to the total count of spattered ions were significantly larger in proliferative diabetic retinopathy than in proliferative vitreoretinopathy. The average ratios of each of vitamin A fragment-1 and palmitic acid ion counts to the total count of spattered ions were significantly larger in proliferative diabetic retinopathy than in proliferative vitreoretinopathy. The fact that diabetic and non-diabetic preretinal proliferative tissues contain organic substances suggests that these proliferative tissues are derived from the retina, especially the deep retina, probably the photoreceptor cell and/or retinal pigment epithelium. The presence of Zn and Cu also suggest the same process. Diabetic proliferative tissues rich in organic substances are thought to grow more than do non-diabetic tissues, because vitamins A and E, and fatty acids are necessary for the maintenance of cell structure and the proliferation of tissue. Analysis of biological specimens by TOF-SIMS is expected to provide valuable information. PMID- 12892226 TI - Support film in transmission electron microscopy: experiences with polyvinyl butyral Pioloform BM 18. AB - Microscopic work with single-slot grids requires high-quality support films to span the relatively large gap. The imminent unavailability of the polyvinyl formal Pioloform FN 65, which to date has been used as the standard polyvinyl formal for the generation of support films in transmission electron microscopy (TEM), has necessitated the finding of a substitute material to produce such films. Therefore, we compared the polyvinyl butyral Pioloform BM 18 with the polyvinyl formal Pioloform FN 65 for the production of TEM support films, using operational criteria for assessment. Pioloform BM 18 with the solvent chloroform resulted in support films of unacceptable quality compared with Pioloform FN 65. Adding the softener dibutyl phthalate to the chloroform solvent for Pioloform BM 18 markedly improved the film quality, resulting in support films with high transparency and flexibility, and even greater stability in the electron beam when compared with films of Pioloform FN 65. Pioloform FN 65 also had the disadvantage of requiring highly toxic 1,2-dichloroethane as a solvent, whereas Pioloform BM 18 can be used with chloroform. PMID- 12892227 TI - Payment of research subjects involved in clinical trials is unethical. PMID- 12892228 TI - Imaging of experimental rat gliomas using a clinical MR scanner. AB - BACKGROUND: Studies of brain tumor development in experimental animal models have to date mostly been based on post-mortem histological examinations. The use of magnetic resonance imaging (MRI) may provide a non-invasive technique for studying tumor growth and treatment effects in such animal models. However, most of these studies have been performed on purpose-dedicated small bore magnetic resonance (MR) systems, of high cost and limited availability. The purpose of this study was thus to obtain high-resolution images of experimental gliomas in the rat brain, using a clinical 1.5 T MR scanner. METHODS: Anesthesized rats bearing BT4C brain tumors were positioned into a specially designed immobilizing device, and a small circular coil was positioned onto the skulls. Two T1 weighted series were acquired before and after subcutaneous contrast injections. A T2 weighted series was also obtained. The rats were then sacrified, the brains removed, and the histological tumor volumes were compared to the volumes obtained on MRI. RESULTS: There were visible tumors in 10 of 13 animals scanned on MR. The rim of the tumors were visualized on T1 weighted series without contrast. On T1 images with contrast, the tumors were seen as high signal intensity areas. The T2 weighted images showed peritumoral edema. No necrosis or cystic parts of the tumors were detected. There was a consistency between the MR and the histology findings, showing a high degree of correlation between the two volume determination methods. CONCLUSIONS: High-resolution images of experimental rat gliomas can be obtained using a clinical MR scanner and a commercially available RF coil. This MRI technique may also be expanded to extraneural rat tumor models, for studies of tumor development and treatment. PMID- 12892229 TI - Preoperative proton-MR spectroscopy of gliomas--correlation with quantitative nuclear morphology in surgical specimen. AB - A comparison between data from proton-MR spectroscopy (1HMRS) and quantitative histomorphology of tumor cell nuclei in gliomas has not been reported up to now. Therefore, the question must be answered, if there are any significant correlations between histomorphology of gliomas and quantitative data from 1HMRS concerning tissue metabolites. Surgical glioma specimen (glioblastomas, astrocytomas, oligodendrogliomas) from 46 patients with tumor grades II-IV according to WHO have been evaluated by means of a digital image analysis system using Ki-67-immunostained paraffin sections. Nuclear density, Ki-67-proliferation index, nuclear area and shape variables (roundness factor, Fourier-amplitudes) have been determined from 200 randomly selected tumor cell nuclei in each tumor specimen. These data have been correlated with preoperative data from 1HMRS. A positive correlation between Fourier-amplitudes, choline peak and lipide peak was observed, as well as a negative correlation between these variables and the nuclear roundness factor. This result indicates higher choline and lipide peaks with increasing irregularity of nuclear outlines. Proliferation index Ki-67 was positively correlated with the lipide peak, nuclear density showed a positive correlation with the choline peak. Glioblastomas (n = 29) showed an additional positive correlation between mean nuclear size and total creatine. Anaplastic gliomas (n = 12) showed a positive correlation between lactate peak and the standard deviation of the nuclear roundness factor. Further multivariate analyses have shown, that for the present collective of 46 cases, histometric variables have a higher significance than spectroscopic data for the differentiation of the different tumor grades. These results verify a significant correlation between preoperative data from 1HMRS and histomorphology of tumor cell nuclei in gliomas, supporting the biological significance of both histomorphometry and 1HMRS for the evaluation of these tumors. PMID- 12892230 TI - Radioimmunoscintigraphy of intracranial glioma xenograft with a technetium-99m labeled mouse monoclonal antibody specifically recognizing type III mutant epidermal growth factor receptor. AB - The type III mutant epidermal growth factor receptor (EGFR) is expressed on the cell surface of a subset of glioma, but not of normal tissues. In this study, we investigated the in vivo kinetics of 3C10 mouse monoclonal antibody (mAb), specifically recognizing the type III mutant EGFR (EGFRvIII), using athymic nude mice bearing the intracranial glioma xenograft overexpressing the EGFRvIII. Human glioma cell line, U87MG, expressing the wild type EGFR and the transfectant, named U87MG x deltaEGFR, expressing the EGFRvIII, were transplanted subcutaneously or intracranially to nude mice. 3C10 mAb labeled with a technetium 99m (99mTc) was intravenously injected into these nude mice and then the mice were sacrificed at 24 h later, and the 99mTc-uptake by xenografts and major normal organs was measured to determine the biodistribution of mAb. Furthermore, at 3, 6 and 24 h after injection of 99mTc-labeled 3C10 mAb, whole-body scintigraphy was obtained with a gamma camera to localize the tumor site. 3C10 mAb significantly accumulated to U87MG x deltaEGFR xenografts transplanted subcutaneously or intracranially in nude mice, showing high tumor-to-blood ratio of 10.30 and 4.01, respectively. In contrast, uptake of control antibody in the intracranial tumor was as low as 0.43. In scintigrams, intracranially transplanted U87MG x deltaEGFR xenografts were detectable at 3 h after injection of 99mTc-labeled 3C10 mAb. These results suggest that intravenously injected 3C10 mAb specifically accumulated to the subcutaneous or intracranial glioma xenograft expressing the EGFRvIII and 3C10 mAb is a potential diagnostic and therapeutic agent for patients with gliomas expressing the EGFRvIII. PMID- 12892231 TI - Molecular analysis of the rhabdoid predisposition syndrome in a child: a novel germline hSNF5/INI1 mutation and absence of c-myc amplification. AB - The authors report a case of the rhabdoid predisposition syndrome (RPS) secondary to a germline hSNF5/INI1 mutation, whose brain tumor was originally unclassified but finally diagnosed as an atypical teratoid/rhabdoid tumor (AT/RT) by molecular analysis. A 7-month-old infant presented with hydrocephalus secondary to a huge pineal tumor and subsequently developed a renal rhabdoid tumor. The histology of the brain tumor was initially undetermined; however, an AT/RT was strongly suspected because of her clinical course. Mutational screening of the hSNF5/INI1 gene by heteroduplex and direct sequence analysis detected a missense mutation at codon 53 (CGA --> TGA, arginine --> stop) in both tumors, as well as in normal tissue of the kidney. Polymerase chain reaction (PCR)-based microsatellite analysis showed in both tumors allelic loss on chromosome arm 22q to which the hSNF5/INI1 gene maps. c-myc amplification was examined by differential PCR but not detected. Histologic review of the brain tumor by immunohistochemistry confirmed focal expression of epithelial membrane antigen and smooth muscle actin. These findings suggest that the brain tumor was really an AT/RT as a component of RPS secondary to a germline hSNF5/INI1 mutation. The present mutation has never been reported in the literature. PMID- 12892232 TI - Pneumocystis carinii pneumonia in HIV negative patients with primary brain tumors. AB - BACKGROUND: Pneumnocystis carinii pneumonia (PCP) is an opportunistic infection with rather adverse outcomes. An unexpected increase in cases of PCP was noted in the brain tumor population at the Johns Hopkins Hospital (JHH) in 2000. This prompted the present review of the clinical features and risk factors for PCP in the human immunodeficiency virus (HIV) negative brain tumor population. METHODS: The study was located at the JHH. A retrospective review of medical records was done to identify patients with discharge diagnosis of PCP from 1980 to 2001. Patients who were HIV positive were excluded. A detailed analysis was done of patients with brain tumors. RESULTS: From 1980 to 2001, 468 cases of PCP were identified, diagnosed histologically or clinically, of which 110 were patients with an underlying malignancy. Of the 110 cases 15 were seen in the brain tumor population. Of these, 6 patients were seen in 2000 and one in early 2001. Three of these had primary central nervous system (CNS) lymphoma (PCNSL) on high dose methotrexate. Eight of the fifteen episodes (53.3%) were fatal despite institution of antibiotics and supportive therapy. CONCLUSION: The incidence and mortality due to Pneumocystis carinii among the brain tumor population is increasing. While corticosteroids are known immunosupressants, prescribing patterns for these medications has not changed lately. However, high dose methotrexate is now being used in PCNSL and could be a complicating factor. Since effective prophylaxis exists, it should be considered in patients with brain tumors receiving high dose steroids, high dose methotrexate or with lymphopenia. PMID- 12892233 TI - Safety of multiple stereotactic radiosurgery treatments for multiple brain lesions. AB - BACKGROUND: Stereotactic radiosurgery (SRS) is a widely used therapy for multiple brain lesions, and studies have clearly established the safety and efficacy of single-dose SRS. However, as patient survival has increased, the recurrence of tumors and the development of metastases to new sites within the brain have made it desirable to repeat treatments over time. The cumulative toxicity of multi isocenter, multiple treatments has not been well defined. We have retrospectively studied 10 patients who received multiple SRS treatments for multiple brain lesions to assess the cumulative toxicity of these treatments. METHODS: In a retrospective review of all patients treated with SRS using the X-knife (Radionics, Burlington, MA) at Westchester Medical Center/New York Medical College between December 1995 and December 2000, 10 patients were identified who received at least two treatments to at least 3 isocenters and had a minimum follow-up period of 6 months. Image fusion technique was used to determine cumulative doses to targeted lesions, whole brain and critical brain structures. Toxicities and complications were identified by chart and radiological review. RESULTS: The average of the maximum doses (cGy) to a point within the whole brain was 2402 (range 1617-3953); to the brainstem, 1059 (range 48-4126); to the right optic nerve, 223 (range 14-1012); to the left optic nerve, 159 (range 17-475); and to the optic chiasm, 219 (range 15-909). There were no focal neurological toxicities, including visual disturbances, cranial nerve palsies, or ataxia in any of the 10 patients. There were also no global toxicities, including cognitive decline or secondary tumors. Only one patient developed seizures that were difficult to control in association with radiation necrosis. CONCLUSIONS: Multiple SRS treatments at the cumulative doses used in our study are a safe therapy for patients with multiple brain lesions. PMID- 12892235 TI - Tolerance of nitrosurea-based multiagent chemotherapy regime for low-grade pediatric gliomas. AB - The aim of this study was to compare tolerance of a nitrosurea-based regime with 'standard' therapy of vincristine (VCR) and carboplatin for low-grade gliomas. Ten children with low-grade gliomas received second line therapy consisting of thioguanine, procarbazine, CCNU and vincristine (TPCV). Two groups were identified, i.e. patients who had either experienced significant toxicity with carboplatin (reaction group) or had re-growth of their tumor (re-growth group) following first line therapy. Patients were evaluated for toxicity. Data was available on nine patients. Patients in the reaction group completed a mean of 3 cycles of TPCV (range 2-4). One patient stopped after 2 cycles of TPCV due to tumor progression and died 3 months later and one remained on therapy at the time of analysis. Patients in the re-growth group received a mean of 5.5 cycles of TPCV (range 4-8). Treatment was discontinued in one patient after 4 cycles due to hematological toxicity, one experienced tumor progression after 4 cycles and one stopped after 6 cycles because of neurological toxicity. There was no difference in the incidence of grade 3/4 neutropenia or thrombocytopenia, transfusion requirements or delays in chemotherapy between TPCV and VCR/carboplatin in either group. There were no serious infections or toxic deaths. Seven of nine patients had stable disease at a mean of 13 months of follow up. TPCV therapy is a well tolerated regime with comparable bone marrow toxicity to VCR/carboplatin. Significant disease stabilization was observed with TPCV and hence this regime may be used as second line therapy. PMID- 12892234 TI - Psychosocial and behavioral functioning among pediatric brain tumor survivors. AB - OBJECTIVE: To describe the psychosocial and behavioral functioning, as described by patient, parent and teacher, of a cohort of adolescents who have been previously treated for a brain tumor. METHODS: A cohort of 32 patients, 12-18 years old, were evaluated between 1 and 5 years post-treatment for brain tumor during the patient's regularly scheduled follow-up clinic appointment at the Dana Farber Cancer Institute. The Self-Report questionnaire and the Parent-Report of the Behavioral Assessment System for Children (BASC) were administered to the patient and to one of the patient's parents, respectively. In addition, the BASC Teacher-Report was completed by the patient's teacher. Descriptive statistics were generated; binomial distribution analyses were carried out to assess whether the proportion of individuals with impaired performance on each measure exceeded normative expectations. RESULTS: Comparison of the proportion of patients with elevated scores to normative expectations indicated no excess of elevated scores on any of the BASC scales of the Self-Report. However, parents endorsed items in the areas of attention problems and leadership; teachers endorsed items concerning learning problems; and both parents and teachers endorsed items indicative of somatization behaviors. CONCLUSIONS: Parent and teacher feedback indicate some level of psychosocial and behavioral morbidity for adolescents treated for a brain tumor; this finding contrasts with adolescent Self-Report indicating no difficulties in behavioral and psychosocial functioning. The extent to which these vulnerabilities impact quality of life and the discrepancy between reporters should be assessed in follow-up studies with a larger cohort of patients. PMID- 12892236 TI - Leptomeningeal metastases from recurrent squamous cell cancer of the skin. AB - We report two patients with leptomeningeal metastases from squamous cell cancer of the skin. In one patient cauda equina metastases appeared 6 years after initial presentation; the tumor from radical neck dissection demonstrated perineural invasion suggesting a possible route for subarachnoid dissemination. The other patient presented 2 years after initial diagnosis with confusion and cranial nerve seven and twelve palsies. Both patients died from complications of leptomeningeal metastases. PMID- 12892237 TI - Primary leptomeningeal primitive neuroectodermal tumor. AB - Leptomeningeal metastases are a common complication of medulloblastomas and other primitive neuroectodermal tumors (PNETs). Much rarer are PNETs apparently arising in the leptomeninges. An 8-year-old boy presented with headache and vomiting, due to neoplastic meningitis from primary neuroectodermal tumor without an identifying mass. After craniospinal irradiation and chemotherapy (carboplatin, vincristine, cyclophosphamide and lomustine) the boy was in remission for 14 months, then suffered several relapses despite various chemotherapy regimens. After the initial presentation of 3.5 years the boy began to suffer from prolonged refractory non-convulsive status epilepticus and later expired from progression of primary leptomeningeal PNET. PMID- 12892238 TI - Role of early radiotherapy in the treatment of supratentorial WHO Grade II astrocytomas: long-term results of 97 patients. AB - OBJECT: To determine principal prognostic factors and the effect of timing of radiotherapy (RT) on disease-specific survival (DSS) and progression-free survival (PFS) in WHO Grade II astrocytomas. METHODS: Histologic slides of 166 consecutive patients with the original tissue diagnosis of low-grade, non pilocytic astrocytoma were reviewed. One-hundred and six were selected where two additional certified neuropathologist agreed on the grading of WHO Grade II astrocytoma. In 97 out of 106 cases follow-up informations were available. Early postoperative RT was given to 36 out of 97 patients (37%). The two groups of patients (early vs. delayed RT) were well balanced in respect to extent of surgery and other main clinical prognostic factors. Median follow-up of surviving patients was 79 months. The 5- and 10-year PFS was 52.2% and 30.7% with early RT and 39.5% and 12.4% with delayed RT (p = 0.0388). In respect to DSS, there was no significant difference in the 5- and 10-year actuarial survival rate according to the timing of RT (60.5% and 26.5% vs. 66.6% and 23.7%; p = 0.7545). Age (p = 0.0145) and extent of surgery (p = 0.0473) were significant prognostic variables in respect to DSS. Subdividing the irradiated group based on the extent of surgery, early RT in the subtotal group significantly improved 5-year PFS (60.0% vs. 12.4%; p = 0.0036) and DSS (66.7% vs. 49.8%; p = 0.0389). However, postoperative RT had no influence on PFS (p = 0.6812) and DSS (p = 0.3987) in the group with extensive resection. CONCLUSION: Early postoperative RT in subtotally resected, Grade II astrocytomas significantly improves both progression-free and disease-specific survival. Early RT does not benefit patients with extensive resection, RT should be withheld in these patients until progression. PMID- 12892239 TI - Nervous system involvement by pancreatic cancer. AB - From 1,229 consecutive patients with pancreatic cancer, seven patients who developed nervous system metastasis were evaluated retrospectively. Nervous system metastasis frequently antedated the diagnosis of the primary cancer (five out of seven patients). Four patients had cerebral metastasis. Brain computed tomography or magnetic resonance imaging revealed single or multiple cyst-like lesions with ring enhancement in half of these patients. Three patients had spinal metastasis which caused myelopathy in two. The metastatic lesions were localized to the thoracic level. Radiation therapy directed to the metastatic sites could not change the clinical course. PMID- 12892240 TI - Cancer rehabilitation: particularly with aspects on physical impairments. AB - Cancer can cause multiple impairments, activity limitations and participation restrictions. According to individual case findings and needs, rehabilitation treatment is varied. The review mainly focuses on specific problems. Because of functional deficits cancer patients suffer from persistent emotional and social distress and a reduced quality of life (QOL). QOL encompasses at least the four dimensions of physical, emotional, social and cognitive function, which may be positively influenced by physical exercise. Physical exercise also has been shown to prevent or minimise inactivity/ disuse problems and to reduce fatigue. The management of sexuality dysfunction has to begin with a thorough history taking and a consequent sexuality counselling. The goals of rehabilitation procedures under palliative care are not only to control physical pain but also to help with mental, social and spiritual pain, together with other symptoms. Rehabilitation problems in head and neck cancer, sexuality, lung cancer, prostate cancer, breast cancer and lymphedema can be improved by rehabilitation. The review mainly focuses on impairment and activity limitation. Social, psychological and vocational aspects are left aside in this review. PMID- 12892241 TI - Efficacy of dynamic lumbar stabilization exercise in lumbar microdiscectomy. AB - OBJECTIVE: The aim of this study was to determine the efficacy of dynamic lumbar stabilization exercises in patients with lumbar microdiscectomy. DESIGN: A prospective, randomized, controlled study. SUBJECTS: Forty-two patients who were diagnosed as having lumbar disc herniation and had been operated on using the microdiscectomy method were divided randomly into 3 groups. METHODS: Dynamic lumbar stabilization exercises were set for the first group and a home exercise programme for the second. The third group given no exercises was considered as a control group. All patients were examined twice, once before the exercise programme and once 8 weeks later. RESULTS: Improvement in the first group was highly significant after the treatment (p < 0.0001). The second group improved significantly more in some parameters (pain, functional disability, lumbar Schober, progressive isoinertial lifting evaluation (neck), trunk endurance (flexion-extension)) than did the third group. The third group of patients showed some improvement in fingertip-floor distance, functional disability, modified lumbar Schober and left rotation in 8 weeks, but there were no significant improvements in the other parameters. CONCLUSION: Dynamic lumbar stabilization exercises are an efficient and useful technique in the rehabilitation of patients who have undergone microdiscectomy. They relieve pain, improve functional parameters and strengthen trunk, abdominal and low back muscles. PMID- 12892242 TI - Course of back pain in primary care: a prospective study of physical measures. AB - OBJECTIVE: To describe physical measures used in patients with back pain when no specific treatment is given, to examine associations between change over time in these measures and changes in pain and back-related disability, and to study the value of physical measures at baseline and at a 4-week follow-up to predict outcome at 12 months. DESIGN: A prospective consecutive study. SUBJECTS: Forty four patients presenting with low back pain in primary care. METHODS: The patients underwent a physical examination at baseline and at 4 weeks. Follow-up was carried out using questionnaires until 12 months. Linear regression was used to identify predictors. RESULTS: Most measures had improved significantly at the 4-week follow-up. Thoracolumbar rotation, isometric endurance back extensors, and fingertip-to-floor distance at 4 weeks were significant predictors for pain intensity and back-related disability at the 12-month follow-up. Eighteen out of 44 patients reported an increase in pain after the assessment of the physical measures at baseline. This group of patients improved more in physical measures between baseline and the 4-week follow-up. CONCLUSION: Physical measures assessed at the 4-week follow-up, but not at baseline, could provide important additional information for identifying those patients at risk for worse outcome in pain or back-related disability at 12 months. PMID- 12892243 TI - Recovery after total hip joint arthroplasty in elderly patients with osteoarthritis: positive effect of upper limb interval-training. AB - OBJECTIVE: To evaluate the influence of an interval training program for the upper limbs on cardiorespiratory fitness and walking ability in elderly patients after total hip joint arthroplasty. DESIGN: A randomized controlled trial. After surgery, control and training groups started general rehabilitation. Training group combined it with an arm-interval exercise program (3 sessions of 30 minutes per week, for 6 weeks). SUBJECTS: Patients were assigned randomly to control (n = 7) and training groups (n = 7). METHODS: Incremental exercise tests were carried out until exhaustion on an arm crank ergometer 1 month before and 2 months after surgery. A 6-minute walk test was also performed 2 months after surgery. RESULTS: VO2 peak increased significantly in the training group (p = 0.0424) and did not change in the control group. The difference in VO2 peak change between the groups was significant (p = 0.0362, +19.2% in the training group and -3.5% in the control group). In the walking test the training group covered a significantly longer distance than did the control group (p = 0.0055, 396.4 metres and 268.1 metres, respectively). CONCLUSION: These results stress the importance of physical training in a rehabilitation program after total hip joint arthroplasty and this should be considered for improving the current practices in rehabilitation. PMID- 12892244 TI - Sitting up and transferring to a chair: two functional tests for patients with stroke. AB - OBJECTIVE: The aim of the study was to evaluate the reliability of two clinical tests of mobility for stroke patients. DESIGN: The study took place in a university hospital with a case control design. SUBJECTS: A sample of convenience including 31 subjects: a group of patients with prior stroke (n = 19); a control group (n = 12). METHODS: The time(s) for sitting up from a supine position and transferring from an examination table to a chair were measured and inter- and intra-rater reliability were assessed. The source of variation in the test results as well as intra-class correlations were estimated. RESULTS: The largest source of variation in the supine position was between subjects, and the between tester variability was very low, with similar results for the transfer from table to chair. The intra-class correlations are all high (range 0.77-0.98). This indicates that it is of little importance which tester is performing the tests. CONCLUSION: Sitting up on an examination table and transferring from an examination table to a chair seem to be reliable timed clinical tests of mobility in stroke research. PMID- 12892245 TI - Trunk muscles in persons with hemiparetic stroke evaluated with computed tomography. AB - OBJECTIVES: To analyse side difference in bilateral trunk muscles in patients with hemiparetic stroke, to relate it with impairment and disability variables and to evaluate longitudinal changes. METHODS: In a sample of 83 inpatients with hemiparetic stroke undergoing rehabilitation, we measured the cross-sectional area of the paravertebral muscle and thigh muscles using computed tomography at admission and discharge. Classifying them by paravertebral muscle side difference (group I: contralateral > ipsilateral; II: contralateral = ipsilateral; III: contralateral < ipsilateral) we analysed group difference in the Stroke Impairment Assessment Set, the Functional Independence Measure and walk velocity. RESULTS: In contrast to thigh muscles, the paravertebral muscle cross-sectional area was significantly greater on the side contralateral to the brain lesion. Discharge paravertebral muscle cross-sectional area increased significantly from admission values. The Stroke Impairment Assessment Set, Functional Independence Measure and walk velocity were significantly lower in group I. CONCLUSION: The contralateral paravertebral muscle cross-sectional area was larger than the ipsilateral ones, and this was related to the degree of impairment and functional limitations. PMID- 12892246 TI - Test-retest intra-rater reliability of grip force in patients with stroke. AB - OBJECTIVE: Coefficients of repeatability and reproducibility can be guides in differentiating between real changes and measurement error. The aim was to evaluate test-retest intra-rater reliability of a clinical procedure measuring grip force with Grippit in stroke patients, to assess relationship between grip force of the hands and between sustained and peak grip force. PATIENTS AND METHODS: Eighteen patients were tested using the Grippit at two occasions one hour apart. Each occasion comprised three consecutive trials per hand. RESULTS: The paretic hand needs to score a 50 N change within and between occasions to exceed the measurement error in 95% of the observations, irrespective of calculation method. Expressed by CV(within) the measurement error was 10%. There was no learning or fatigue effect during measuring. There was a wide variation between subjects but the mean ratio between sides was 0.66. The mean ratio between sustained and peak grip force was 0.80-0.84. CONCLUSION: The measurement errors were acceptable and the instrument can be recommended for the use in stroke patients at a department of rehabilitation medicine. PMID- 12892247 TI - Functional recovery after concomitant fractures of both hip and upper limb in elderly people. AB - OBJECTIVE: To investigate functional recovery after concomitant fractures of both hip and upper limb in elderly people. DESIGN: Survey study. SETTING: Rehabilitation hospital in Italy. PARTICIPANTS: 586 consecutive in-patients with hip fracture. MAIN OUTCOME MEASURES: Functional recovery was evaluated by using Barthel index score. RESULTS: 4.1% of patients (i.e. 24/586) suffered from a concomitant fracture of an upper limb, involving proximal humerus (n = 8) or distal radius (n = 16). After adjustment for 9 prognostic factors, a significant reduction in the Barthel index score on admission but not on discharge was found in the patients with an upper limb fracture. The length of stay was not significantly associated with the presence of the concomitant upper limb fracture. CONCLUSIONS: In a sample of hip-fractured patients, neither the functional recovery at the end of a course of rehabilitation nor the length of stay were influenced by the presence of a concomitant fracture involving an upper limb. PMID- 12892248 TI - Magnetic resonance imaging of the fifth metatarsophalangeal joint compared with conventional radiography in patients with early rheumatoid arthritis. AB - OBJECTIVE: To evaluate if magnetic resonance imaging (MRI) is superior to conventional radiography for detection of erosions in the fifth metatarsophalangeal (MTP5) joint. METHODS: Within one year from the onset of rheumatoid arthritis (RA) (baseline), one and three years thereafter MRI and conventional radiographs of the MTP5 joint were performed in 23 patients. RESULTS: MRI revealed erosions in 10 patients at baseline, in 15 after one year and in 15 patients after 3 years. On conventional radiography, there were erosions in 10 patients at baseline, 16 after one year as well as after 3 years. The agreement between the two imaging methods was fair to good at baseline and after one and three years (kappa 0,65, 0,51 and 0,51 respectively). The number of patients with clinical evidence of synovitis decreased considerably over time although the number of patients with MRI-synovitis was unchanged and the number of patients with erosions increased. CONCLUSIONS: MRI was not superior to conventional radiography in detecting erosions in MTP5 joints in patients with early RA. Most erosions developed during the first year of observation. Synovitis on MRI may be a marker of future development of erosions in the MTP5 joint. PMID- 12892249 TI - Muscle metabolism in fibromyalgia studied by P-31 magnetic resonance spectroscopy during aerobic and anaerobic exercise. AB - OBJECTIVE: To investigate mechanisms underlying the reduced work capacity of fibromyalgia (FM) patients were compared to healthy controls at specified workloads, using P-31 magnetic resonance spectroscopy (MRS). METHODS: The forearm flexor muscle group was examined with MRS at rest, at sub maximal and at maximal controlled dynamic work as well as at maximal isometric contraction. Aerobic fitness was determined by bicycle ergonometry. RESULTS: Metabolite concentrations and muscle pH were similar for patients and controls at lower workloads. At maximal dynamic and static contractions the concentration of inorganic phosphate was lower and at static contractions the pH decrease was smaller in patients. The performed work by patients was only 50% compared to controls and the patients experienced more pain. Maximal oxygen uptake was lower in the fibromyalgia group. Expired gas-analysis in this group showed ventilatory equivalents at similar relative levels of maximal work capacity. CONCLUSION: Fibromyalgia patients seem to utilise less of the energy rich phosphorous metabolites at maximal work despite pH reduction. They seemed to be less aerobic fitted and reached the anaerobic threshold earlier than the controls. PMID- 12892250 TI - Bone mineral density in fibromyalgia patients--correlation to disease activity. AB - OBJECTIVE: To compare bone mass (BMD) in women with fibromyalgia (FM) with healthy females, and to evaluate whether self-reported pain and lack of functional capacity correlate to reduced BMD in FM patients. METHODS: Thirty-one FM patients (20 pre- and 11 postmenopausal) and fourty-one healthy women (30 pre- and 10 postmenopausal) were enrolled in the study. BMD of the lumbar spine and the femoral neck was measured by a DEXA (Norland) scanner. Self reported pain was measured on a Visual Analog Scale (VAS). The Activity of Daily Living (ADL) component of the Fibromyalgia Impact Questionnaire (FIQ-ADL) was used as measure for physical capacity. RESULTS: BMD-lumbar spine and BMD-femoral neck did not differ significantly between FM patients and controls, though premenopausal FM patients tended to have lower BMD-femoral neck (p = 0.09). Self reported pain and FIQ-ADL among FM patients correlated with BMD-femoral neck (r(s) = -0.52, p = 0.003); (r(s) = -0.31, p = 0.09). CONCLUSION: Premenopausal FM patients tended to have lower BMD of hip than controls. Self reported pain correlated negatively to BMD. Thus, the severity of FM might have a negative impact on bone mass. PMID- 12892251 TI - Profiles of the acute-phase reactants C-reactive protein and ferritin related to the disease course of patients with systemic lupus erythematosus. AB - OBJECTIVE: To obtain insight in the acute-phase response in SLE. METHODS: The clinical history, SLEDAI, CRP and ferritin concentrations were analysed throughout the disease course of 10 SLE patients. RESULTS: During a mean follow up of 4.8 years, 10 exacerbations (SLEDAI > or = 11) occurred. Throughout the disease course, CRP and SLEDAI correlated positively in 5 patients, whereas the correlation between SLEDAI and ferritin was positive in 7 patients. However, elevated CRP concentrations together with elevated ferritin levels were only observed during 4 exacerbations. Ferritin concentrations were exceptionately high (> 1500 microg/L) during 4 flare-ups. CRP and ferritin levels remained normal during 5 exacerbations. CONCLUSION: SLE is characterised by highly variable and unusual CRP and ferritin responses that do not always reflect the extent of inflammation in individual patients. Despite severe disease activity, ferritin levels can remain well within the normal range, limiting its clinical usefulness as a marker for disease activity. PMID- 12892252 TI - Serum levels of cartilage oligomeric matrix protein. A predicting factor and a valuable parameter for disease management in rheumatoid arthritis. AB - OBJECTIVE: To examine whether cartilage oligomeric matrix protein (COMP) correlates with inflammation and/or joint destruction of patients with rheumatoid arthritis (RA) and to test COMP as predicting factor for the outcome of patients with established RA. METHODS: Serum levels of COMP were measured in sera of 62 patients, suffering from RA according to the ACR criteria and treated in intervals in our department, over a period of 5 years. A commercially available sandwich--type ELISA-kit developed by AnaMar Medical AB, Sweden, was used. The results of serum COMP were compared with the Disease Activity Score (DAS), the Larsen Score, and clinical and laboratory parameters. RESULTS: We found a positive correlation between serum levels of COMP at baseline and deterioration of Larsen score even after 5 years (p < 0.007; r = 0.34). To confirm serum COMP as an independent predicting factor for patients with RA we looked at a subgroup of patients (n = 17) with elevated serum levels of COMP (mean 11,7 U/l) and low clinical prognostic factors. In this subgroup we also found a significant correlation with delta Larsen score (p < 0.01; r = 0.59) after 5 years. CONCLUSION: Serum levels of COMP is known to reflect increased cartilage turnover. The results indicate that serum COMP may be used as a prognostic marker of cartilage degradation in a patient group with established RA. PMID- 12892253 TI - Gastro-protective treatment in patients using NSAIDs. Development of appropriateness criteria by a multidisciplinary expert panel. AB - OBJECTIVE: To refine the appropriate indications for gastro-protective treatment in patients using non-steroidal anti-inflammatory drugs (NSAIDs). METHODS: Using the RAND Appropriateness Method, a multidisciplinary expert panel was asked to rate the appropriateness of gastro-protection for 4608 different cases. Logistic regression was used to construct a decision framework to identify patients for whom gastro-protective medication should be considered. RESULTS: Complete consensus existed on the appropriate use of gastro-protection in patients with a history of (un)complicated peptic ulcer. For other patients. agreement was found for 39% over 1536 cases. Logistic regression demonstrated strong consistency of the panel ratings (Hosmer Lemeshow coefficient 0.92), allowing the development of a comprehensive decision support model. CONCLUSIONS: Using the RAND Appropriateness Method, we were able to develop a clear and internally consistent decision framework for the appropriate use of gastro-protection in patients taking NSAIDs. The validity of this model should be tested in further studies and practice. PMID- 12892254 TI - Mandibular osteotomies in patients with juvenile rheumatoid arthritic disease. AB - OBJECTIVE: This retrospective study evaluates the results after orthognathic surgery in a group of patients with juvenile rheumatoid arthritis. METHODS: The material comprised sixteen patients where genioplasty with or without bilateral sagittal split (BSSO) had been performed during a 10-year period between 1991 and 2000. The patients were recalled for follow-up examination and the clinical records and radiographs of the patients were analysed. RESULTS: All patients reported an improved facial esthetics. Sixty-two% reported altered neurosensory dysfunction in the inferior alveolar nerve, but no patients reported altered feelings to interfere with function. Two patients reported reduction in pain in the TMJ from a score 10 and 7 in the VAS-scale preoperatively, to 0 after the orthognathic surgery. Eight of the patients reported this to be less uncomfortable compared to other surgical procedures because of their JRA. Ninety four% noted a positive social change after the operation. CONCLUSION: Orthognathic surgical treatment of the JRA patient improves the facial profile. The described procedures are safe and serious complications were not seen in our study. PMID- 12892255 TI - Relationship between functional ability and physical fitness in juvenile idiopathic arthritis patients. AB - OBJECTIVE: To determine the relationship between aerobic and anaerobic physical fitness and functional ability in children with juvenile idiopathic arthritis (JIA). METHODS: Eighteen children with JIA (age 7 to 14 yr., 3 male/15 female) performed a maximal aerobic exercise test and a Wingate anaerobic exercise test. Functional ability was concurrently assessed using the Childhood Health Assessment Questionnaire (CHAQ). RESULTS: A low relationship between aerobic fitness and functional ability was found (r = 0.0 to 0.4, p > 0.05, except for eating r = 0.46, p < 0.05). The correlations between anaerobic physical fitness and functional ability in JIA patients were strong (r = 0.5 to 0.75, p < 0.05). This indicated a good relationship between anaerobic fitness and functional ability. CONCLUSION: The strong association between anaerobic physical fitness and functional ability showed the importance of anaerobic physical fitness for children with JIA. PMID- 12892256 TI - Insufficiency fractures of the sternum. AB - Insufficiency fractures occur within weakened bones that are unable to withstand the stress of every day normal activities. The spine, pelvis, and lower long-bone extremities are common sites of insufficiency fractures. Spontaneous sternal insufficiency fracture (SIF) has rarely been reported in elderly patients. To recognise a sternal insufficiency fracture is important in selecting the adequate diagnostic procedures. PMID- 12892257 TI - Successful treatment of dermatomyositis-related rapidly progressive interstitial pneumonitis with sequential oral cyclophosphamide and azathioprine. AB - Interstitial lung disease (ILD) is a major complication of idiopathic inflammatory myopathies (IIM). We report the successful use of oral cyclophosphamide, followed by azathioprine maintenance, in the treatment of a patient with dermatomyositis-related rapidly deteriorating ILD, resistant to steroid. Prompt recognition and early aggressive immunosuppressive therapy may improve the outlook of this condition. PMID- 12892258 TI - Coexistence of ankylosing spondylitis and Behcet's disease. Two cases with atypical presentation and course. AB - The prevelance of ankylosing spondylitis or sacroiliitis in Behcet's disease has been the subject of debate for some time. Two cases with atypical presentation and course are discussed. The first patient was a young woman with Behcet's disease whose primary complaint was neck pain. X-ray examination showed severe cervical involvement, characteristic of the type seen in women with ankylosing spondylitis. She subsequently developed anterior uveitis. The second patient was a man suffering from back pain and limited movement of his neck and back. When he was referred to our outpatient clinic, his severe anemia became the subject of the whole investigation. He was diagnosed with Behcet's disease with ankylosing spondylitis accompanied with amyloidosis and an end-stage renal failure. PMID- 12892259 TI - Systemic sclerosis could mask the presentation of psoriasis in a patient with symptomatic and bilateral sacroiliitis. AB - Psoriasis is rarely associated with systemic sclerosis (SSc), and sacroiliitis associated to Connective Tissue Diseases is also rather rare. In this report we describe a case of a young woman with SSc who developed symptomatic and bilateral sacroiliitis. The clinical pattern of sacroiliac involvement of this patient resembles that of psoriatic sacroiliitis. HLA typing was compatible with both SSc and psoriasis. Psoriatic sacroiliitis could not be diagnosed, but, on the basis of these observations, the patient described could represent a case of PsA without psoriasis. and psoriatic cutaneous involvement masked by the presence of SSc. PMID- 12892260 TI - Spastic quadriparesis: an unusual early manifestation of systemic lupus erythematosus. AB - Systemic lupus erythematosus (SLE) is an uncommon immunological disorder with multisystemic involvement. Neurological and neuropsychiatric manifestations in this disease are multifactorial and can involve any part of this system. We describe one patient presenting with spastic quadriparesis as an early clinical manifestation of SLE. This disease should be kept in mind in such a setting, especially if abnormalities in hemogram and urine analysis are seen. Early diagnosis and aggressive therapy may improve the neurological outcome. PMID- 12892261 TI - Intestinal pseudo-obstruction in systemic lupus erythematosus. AB - In order to document intestinal pseudo-obstruction (IPO) as a recently recognized manifestation of systemic lupus erythematosus (SLE), we report the case or a woman with SLE who presented with IPO and we review 21 other previously reported cases from an English literature search. In 41% of the cases, IPO was the initial manifestation of their underlying lupus. The clinical and laboratory features were not significantly different from those reported in large series of patients with SLE, except for an apparent association with an urinary tract involvement (ureterohydronephrosis and interstitial cystitis). The pathogenic mechanism of this complication is not fully understood, but seems to be heterogeneous. IPO responded readily to high dose steroid therapy in all patients, but in some cases this complication evolved regardless of the underlying disease activity. A high level of awareness of this complication is needed to avoid unnecessary surgical intervention. PMID- 12892263 TI - DEFRA outlines its animal health and welfare strategy. PMID- 12892264 TI - Vets to be allowed to dispense other vets' prescriptions. PMID- 12892262 TI - Genetics and susceptibility to RA--more than HLA. PMID- 12892265 TI - Identification of a novel bluetongue virus vector species of Culicoides in Sicily. AB - The vectors of bluetongue virus are certain species of Culicoides biting midges, and in the Mediterranean area Culicoides imicola has long been considered to be the only field vector. In Sicily an entomological and serological surveillance programme has been in operation since the autumn of 2000, which has shown that the prevalence and abundance of C. imicola is lower than in many other Italian regions. Moreover, in 2002, there were outbreaks of bluetongue in the absence of C. imicola, and in these regions bluetongue viral RNA was detected by means of a nested reverse-transcriptase PCR in wild-caught, non-blood-engorged, parous Culicoides pulicaris. Furthermore, bluetongue virus serotype 2 was isolated on five occasions from extracts of non-blood-engorged parous C. pulicaris by using embryonated hens eggs and BHK-21 cells as assay systems. These findings suggest that in parts of Italy and possibly in other areas of Europe, where C. imicola is absent or rare, C. pulicaris may act as a fully competent vector of bluetongue virus. PMID- 12892266 TI - Survey of Campylobacter species, VTEC O157 and Salmonella species in Swedish wildlife. AB - Samples collected from 791 wild animals (Canada geese, roe deer, hares, moose, wild boar and gulls) shot during hunting were examined for verocytotoxin producing Escherichia coli (VTEC) O157, and thermophilic Campylobacter and Salmonella species. With the exception of one positive isolate from a wild boar, VTEC O157 was not isolated from any of the animals. Salmonella species were isolated only from the gulls, of which 4 per cent were estimated to be positive. Thermophilic Campylobacter species were commonly isolated from all the species except deer. PMID- 12892267 TI - Congenital anorectal abnormalities in six dogs. AB - Congenital anorectal abnormalities were diagnosed in three male and three female dogs. One dog had anal stenosis, three had a persistent anal membrane, and the other two had an imperforate anus associated with a rectovaginal fistula. Five of the dogs were treated surgically, and four of them which were followed up for periods ranging from one to five years continued to pass faeces normally. PMID- 12892268 TI - Reovirus-like hepatitis in farmed halibut (Hippoglossus hippoglossus). PMID- 12892269 TI - K-ras mutations in canine pancreatic cancers. PMID- 12892270 TI - Intra-articular injections with high molecular weight sodium hyaluronate as a therapy for canine arthritis. PMID- 12892271 TI - Signet-ring squamous cell carcinoma in a dog. PMID- 12892272 TI - L-2-hydroxyglutaric aciduria in Staffordshire bull terriers. PMID- 12892273 TI - Culling of wild red deer. PMID- 12892274 TI - Supply of POMs. PMID- 12892275 TI - Coprophagy in dogs. PMID- 12892276 TI - Coronoid fractures: operative treatment using an anteromedial approach. PMID- 12892277 TI - Peripheral and central nervous system mechanisms of joint protection. AB - Although there are many risk factors (such as age, family history, and obesity) associated with the development of osteoarthritis (OA), only trauma is known to cause OA. The neuromuscular system controls the amount and kind of movement occurring at a joint, so it is this system that is uitimately responsible for ensuring that joint tissues are not damaged to the point of developing OA during normal day-to-day activities. In the present paper, we review and critically evaluate some of the current concepts of the role of peripheral and central nervous system mechanisms that might protect joints from excessive excursion and joint tissues from excessive loading and OA. We conclude that a neuromuscular protective model based on central pattern generators (CPGs--subconscious motor programs) can best reconcile much heretofore ambiguous information pertaining to the development and progression of OA in stable and unstable joints. PMID- 12892278 TI - Are 2 cages needed with instrumented PLIF? A comparison of 1 versus 2 interbody cages in a military population. AB - Bovine biomechanical data have demonstrated adequate stability of a single threaded interbody cage when combined with a unilateral facet screw during posterior lumbar disc excision and interbody fusion (PLIF). Instrumented PLIF surgery using 1 versus 2 interbody cages was studied in 35 military men with disc disease and chronic low back pain. All patients underwent instrumented PLIF with bilateral diskectomy and partial facetectomy, pedicle screws, autogenous iliac crest bone graft, 1 or 2 interbody cages, and posterior lateral fusion. After an average of 15 months' follow-up, the 2-cage group had a higher rate of dural tear, but rates of other complications, hospital stay, fusion rates, pain levels, functional outcomes, and patient satisfaction were good and did not differ between groups. Costs were higher for the 2-cage group by 1728 dollars per patient. PMID- 12892279 TI - Transforaminal lumbar interbody fusion: unilateral versus bilateral disk removal- an in vivo study. AB - We conducted a study to quantify the attainable cross-sectional area available for fusion and the mean volume of disk removed with a unilateral transforaminal lumbar interbody fusion (TLIF). Starting with the unilateral approach and completing the procedure with a bilateral approach, we removed the maximal amount of disk material. Postoperative computed tomography (CT) images were used to estimate the cross-sectional area/vertebral endplate available for bone grafting and fusion. Results showed that, using the unilateral approach, we removed 11.8 cm3 (range, 7-25 cm3) of disk material--or more than 69% of the total volume of resected disk. Postoperative CT images confirmed that more than 56% of the endplate cross-sectional area is available for fusion using the unilateral approach. We conclude that unilateral TLIF removes sufficient disk material for achieving a solid and stable arthrodesis while minimizing neural retraction and dural exposure. PMID- 12892280 TI - The management of anterior dislocation of the shoulder with proximal humerus fracture, bony Bankart lesion, and associated glenoid labrum avulsion: a report of 2 cases. AB - Proximal humerus fractures are common in the older population. These fractures can be associated with dislocation of the shoulder. Four-part fracture dislocations and head-splitting 3-part fractures are generally indicated for shoulder hemiarthroplasty in the older population. When such surgery is warranted, inspection of the glenoid during the procedure is indicated, as loss of evaluation may lead to postoperative shoulder instability. There is value in alerting orthopedic surgeons to the possibility of anterior glenoid rim fracture in association with proximal shoulder fracture-dislocation. We focus on this issue and advise that the lesion can be visualized and confirmed at open reconstruction and does not need multiple radiographic views. PMID- 12892281 TI - The risks of overly effective postoperative epidural analgesia. AB - Continuous epidural analgesia is frequently used to provide supplemental postoperative pain control. Epidural analgesia has the potential to mask the early symptoms that signal impending complications after even routine surgical procedures. We report a case of sciatic nerve palsy following epidural anesthesia after an uncomplicated leg length correction. Good epidural anesthesia may remove a patient's normal protective sensation, allowing pain and other signs of nerve compression from prolonged unchanged postoperative positioning to go unnoticed. This case highlights the need for heightened awareness of potential neurologic compromise in the setting of epidural analgesia. We recommend closely monitoring the patient's neurologic condition and frequently evaluating the patient's position in bed. PMID- 12892282 TI - Efficacy of telemedicine in the initial management of orthopedic trauma. AB - In order to determine the clinical usefulness of teleradiology in the initial treatment of musculoskeletal injury, a panel of orthopedic trauma surgeons developed a survey that was presented to participants at the annual meeting of the Orthopedic Trauma Association. Selected orthopedic trauma cases from a level 1 trauma and a tertiary care center were presented orally (including a description and interpretation of the radiographic findings) to volunteer participants. Questions related to treatment and patient management were then asked prior to and following the participants' viewing of digital radiographic images. Overall, 90% of respondents found telemedicine useful. They disagreed with the verbal radiographic report 49% of the time. Respondents changed their mind with regard to the need for admission 17% of the time; surgical indications, 22% of the time; and need for more information, 30% of the time. PMID- 12892283 TI - Dynamic hip screw removal using a blade plate extraction device. AB - Dynamic hip screw implants are extracted by removing the side plate and compression screws. Occasionally, ingrown bone will prevent easy plate removal. A common solution is to use an osteotome as a levering device; however, the stress riser on the proximal femur caused by this maneuver could result in a catastrophic fracture. A blade plate extraction device fits into the dynamic hip screw side plate and allows relatively atraumatic removal of the implant, preventing the use of excessive force and decreasing the risk of an intraoperative fracture. PMID- 12892284 TI - Gender and veterinary medicine. PMID- 12892285 TI - An ethicist's commentary on released time for voting, etc. PMID- 12892286 TI - A comparison of the analgesic effects of butorphanol with those of meloxicam after elective ovariohysterectomy in dogs. AB - This study was designed to compare the analgesic effects of butorphanol with those of meloxicam following ovariohysterectomy. Fifteen dogs were premedicated with 0.05 mg/kg body weight (BW) of acepromazine by intramuscular (IM) injection, plus 0.2 mg/kg BW of meloxicam by subcutaneous (SC) injection. Fifteen dogs were premedicated with 0.05 mg/kg BW of Acepromazine, IM, plus 0.2 mg/kg BW of butorphanol, IM. Anesthesia was induced with thiopental, and dogs were maintained on halothane. All pain measurements were performed by 1 experienced individual, blinded to treatment. Pain scores and visual analogue scales (VAS) were performed at 2, 3, 4, 6, 8, 12, and 24 hours postpremedication. An analgesiometer was used to determine the pressure required to produce an active avoidance response to pressure applied at the incision line. Pain scores, VAS, and analgesiometer scores were analyzed by using a generalized estimating equations method. A significance level of P < 0.05 was considered significant. Animals that received meloxicam demonstrated significantly lower pain scores and VAS than did animals that received butorphanol in the first 12 hours after surgery. Results of this study suggest that meloxicam will produce better postoperative analgesia than will butorphanol. Mucosal bleeding times were performed on cooperative animals in the study group (11 butorphanol, 13 meloxicam). Bleeding times were performed prior to premedication, 6 hours following premedication, and 24 hours after premedication. The 6- and 24-hour readings were compared with baseline bleeding times by using a paired t-test with a Bonferroni correction (a significance level of P < 0.025). Bleeding times did not change significantly over time. PMID- 12892287 TI - Growth performance of pigs fed diets with and without tylosin phosphate supplementation and reared in a biosecure all-in all-out housing system. AB - Three hundred and eighty-four pigs, mean initial live weight of 20.8 kg, were assigned randomly to groups of 24 (12 females, 12 castrated males). Each group was randomly assigned to 1 of 2 dietary treatments consisting of the same commercial barley-based diet, with or without the addition of tylosin phosphate. The barn where the animals were housed operates as an all-in all-out facility, and all pigs arrived on the same day as part of a group of 960 pigs. No new pigs were introduced into the facility during the period of this trial and pigs were sent to market over a 4-week period upon achieving a live weight of 110 kg. The pigs were weighed at the beginning of the trial and when they left the facility for slaughter. Feed consumption and incidence of disease, mortality, or both were recorded daily. At slaughter, carcass backfat depth over the last rib, 6.5 cm ventral to the dorsal midline (P-2 site); loin depth; carcass weight; predicted lean yield; and grade index were recorded. The sow herd supplying pigs to the unit was known to be free of the major swine diseases such as swine influenza, mycoplasma pneumonia, porcine reproductive and respiratory syndrome (PRRS), necroproliferative enteritis, and ascarids. A strict biosecurity protocol was employed to minimize the risk of introducing disease organisms into the unit. Prior to this study, no subtherapeutic antibiotics had been used in this facility. Tylosin phosphate supplementation had no significant effect on final weight, days on test, total gain, and daily gain. In both treatments, the pigs reached a mean market weight of 110.2 kg within 94.1 days, resulting in daily gains of the order of 950 grams per day. Due to the design of the trial, it was difficult to measure significant feed consumption effects. Feed consumption and conversion appeared to be similar for pigs in both treatment groups. At slaughter, tylosin phosphate supplementation appeared to significantly increase lean muscle content of the carcass as measured by loin muscle depth (P = 0.04). Mortality rates and the number of underweight pigs sent to market were low for this trial. Mortality was similar for both treatments; however, more of the control pigs than of the tylosin phosphate fed pigs were underweight when sent to market. From the results of this study, it appears that pigs of fast growing genotypes fed adequate diets and housed in a biosecure environment do not require dietary tylosin phosphate supplementation in order to maximize growth. There is some indication that tylosin phosphate supplementation may improve lean content of the carcass in pigs housed in such an environment. PMID- 12892288 TI - A field trial of autogenous Moraxella bovis bacterin administered through either subcutaneous or subconjunctival injection on the development of keratoconjunctivitis in a beef herd. AB - The primary purpose of these experiments was to evaluate an autogenous Moraxella bovis bacterin administered through 2 separate routes of inoculation. An autogenous bacterin was manufactured by using M. bovis recovered from the herd. The bacterin was administered by subcutaneous injection or subconjunctival injection. In each experiment, unvaccinated animals served as controls. Random selection methods were used to place calves into a vaccination or control group. There was no statistical difference in development of infectious keratoconjunctivitis between the vaccinated and unvaccinated calves. There was a statistically significant difference between the sexes; heifers had a higher rate of keratoconjunctivitis. PMID- 12892290 TI - Laparoscopic colopexy in a horse. AB - An 11-year-old Trakehner gelding required 2 ventral midline celiotomies for correction of a large colon volvulus and a large colon displacement, respectively. Laparoscopic colopexy was performed 50 days following the 2nd celiotomy. Delayed laparoscopic colopexy is minimally invasive and does not disrupt the ventral midline incision following abdominal exploration. PMID- 12892289 TI - Efficacy of a paste formulation of omeprazole for the treatment of naturally occurring gastric ulcers in training standardbred racehorses in Canada. AB - The efficacy of a paste formulation of the H+, K+, -ATPase inhibitor omeprazole was evaluated in standardbred racehorses for the treatment and prevention of gastric ulcers. Twenty standardbred racehorses in training, aged 2 to 9 years, were enrolled from 2 training centres in this field trial. Endoscopic examinations confirmed the presence of gastric ulcers in all horses, prior to allocation and treatment and on day 0. Lesions were scored on a scale of 0 to 3 (intact epithelium to extensive ulceration). Replicates were formed, based on training level and location. Within replicates, 1 horse was assigned to group 1 and 3 horses were assigned to group 2, randomly. Horses in group 1 were sham dosed controls. Horses in group 2 were given omeprazole paste orally at 4 mg/kg bodyweight (BW)/day from day 0 to day 27 and 2 mg/kg BW/day of omeprazole paste orally from day 28 to day 57. Follow-up endoscopies were conducted on post treatment days 28 and 58 or 59. Physical examinations, including BWs, were conducted on all horses prior to treatment and on days 13 or 14, 28, 42 or 43, and 58 or 59. Horses treated with omeprazole had significantly (P < 0.01) more improvement in gastric lesion scores than did controls at day 28 and at study termination on days 58 or 59. All of the omeprazole-treated horses were improved relative to baseline ulcer score at both examinations, and 73.3% were healed (lesion score of 0) at both examinations. None of the controls improved at any point during the study. When the dose was reduced to 2 mg/kg BW, 80% of the horses showed no recurrences or worsening in gastric ulcers. It was concluded that omeprazole paste at 4 mg/kg BW orally, once daily is highly effective in healing gastric ulcers in standardbred racehorses in training and that a dose of 2 mg/kg BW orally, once daily, effectively prevents the recurrence of gastric ulcers in most horses. PMID- 12892291 TI - Ovarian hematoma in an 11-year-old Thoroughbred-Hanovarian mare. AB - An aggressive mare, presented for prebreeding examination, was found to have a significantly enlarged ovary (soccer ball size). A granulosa thecal cell tumor was initially suspected. Following laboratory and repeated clinical examination, the mare was diagnosed with an ovarian hematoma, which regressed with treatment. PMID- 12892292 TI - Insulin-secreting beta cell neoplasia in a 10-year-old dog. AB - A 10-year-old collie-cross presented with intermittent episodes of pelvic limb weakness and collapse. Hypoglycemia, secondary to an insulin-secreting beta cell tumor, was suspected on the basis of clinical signs and serum biochemistry. Conservative dietary therapy consisted of multiple small meals, limited exercise, and avoidance of foods containing simple sugars. PMID- 12892293 TI - Osteomyelitis associated with an orthopedic implant. PMID- 12892294 TI - Cutaneous fibropapilloma in a cat (feline sarcoid). PMID- 12892295 TI - Diagnostic ophthalmology. PMID- 12892296 TI - Hunting for evidence in the era of molecular pathology--perils and pitfalls of the problematic chain reaction. PMID- 12892297 TI - Influence of drugs on thyroid function in dogs. AB - Several drugs can affect thyroid function test results in humans and eventually lead to an erroneous evaluation of thyroid function. These medications can alter the synthesis, secretion, transport, or metabolism of thyroid hormones. Some drugs also directly inhibit the hypothalamic-pituitary-thyroid axis. The effects of drugs on thyroid function in dogs have long been underestimated and have most likely contributed to the overdiagnosis of hypothyroidism in this species. This manuscript 1st reviews pertinent thyroid physiology followed by an overview of drugs for which the effects on canine thyroid function have been studied. The effects of glucocorticoids, propranolol, sulfonamides, phenobarbital, potassium bromide, and nonsteroidal anti-inflammatory agents (NSAIDs) on canine thyroid function are summarized here. Knowledge of the potential effect of these medications on thyroid function should contribute to a more reliable interpretation of thyroid function test results in dogs. PMID- 12892298 TI - Structural and functional basis for the long QT syndrome: relevance to veterinary patients. AB - Long QT syndrome (LQTS) is a condition characterized by prolongation of ventricular repolarization and is manifested clinically by lengthening of the QT interval on the surface ECG. Whereas inherited forms of LQTS associated with mutations in the genes that encode ion channel proteins are identified only in humans, the acquired form of LQTS occurs in humans and companion animal species. Often, acquired LQTS is associated with drug-induced block of the cardiac K+ current designated I(Kr). However, not all drugs that induce potentially fatal ventricular arrhythmias antagonize I(Kr), and not all drugs that block I(Kr), are associated with ventricular arrhythmias. In clinical practice, the extent of QT interval prolongation and risk of ventricular arrhythmia associated with antagonism of I(Kr) are modulated by pharmacokinetic and pharmacodynamic variables. Veterinarians can influence some of the potential risk factors (eg, drug dosage, route of drug administration, presence or absence of concurrent drug therapy, and patient electrolyte status) but not all (eg, patient gender/genetic background). Veterinarians need to be aware of the potential for acquired LQTS during therapy with drugs identified as blockers of HERG channels and I(Kr). PMID- 12892299 TI - Association between diabetes mellitus, hypothyroidism or hyperadrenocorticism, and atherosclerosis in dogs. AB - The objective of this study was to determine whether dogs with atherosclerosis are more likely to have concurrent diabetes mellitus, hypothyroidism, or hyperadrenocorticism than dogs that do not have atherosclerosis. A retrospective mortality prevalence case-control study was performed. The study group included 30 dogs with histopathological evidence of atherosclerosis. The control group included 142 dogs with results of a complete postmortem examination, a final postmortem examination diagnosis of neoplasia, and no histopathological evidence of atherosclerosis. Control dogs were frequency matched for age and year in which the postmortem examination was performed. Proportionate changes in the prevalence of diabetes mellitus, hypothyroidism, and hyperadrenocorticism were calculated by exact prevalence odds ratios (POR), 95% confidence intervals (95% CI), and P values. Multiple logistic regression analysis was used to examine the combined effects of prevalence determinants while controlling for age and year of postmortem examination. Dogs with atherosclerosis were over 53 times more likely to have concurrent diabetes mellitus than dogs without atherosclerosis (POR = 53.6; 95% CI, 4.6-627.5; P = .002) and over 51 times more likely to have concurrent hypothyroidism than dogs without atherosclerosis (POR = 51.1; 95% CI, 14.5-180.1; P < .001). Dogs with atherosclerosis were not found to be more likely to have concurrent hyperadrenocorticism than dogs that did not have atherosclerosis (POR = 1.8; 95% CI, 0.2-17.6; P = .59). Diabetes mellitus and hypothyroidism, but not hyperadrenocorticism, are more prevalent in dogs with atherosclerosis compared to dogs without atherosclerosis on postmortem examination. PMID- 12892301 TI - Polypoid cystitis in 17 dogs (1978-2001). AB - Polypoid cystitis is a rare disease of the urinary bladder in dogs characterized by inflammation, epithelial proliferation, and development of a polypoid mass or masses without histopathologic evidence of neoplasia. Medical records of 17 dogs with polypoid cystitis were reviewed to determine the clinical and laboratory features of this disorder and to assess treatment and outcome. Most affected dogs (15/17) were female and presented for evaluation of hematuria or recurrent urinary tract infection (UTI). Proteus spp were the most common bacterial isolates (12/52 or 23.1%) identified when all urine samples obtained for culture at any time during the course study were considered. Other commonly isolated organisms included Escherichia coli , Staphylococcus spp, and Enterococcus spp. Several dogs (7/17) also had cystic calculi at some time during the course of their disease. Most of the masses (11/14) were located cranioventrally in the bladder as opposed to transitional cell carcinoma, which has a predilection for the bladder neck or trigone area. It is unknown whether persistent or recurrent UTI predisposes to polyp formation or if polyps predispose to UTI. Surgery and removal of all polyps was the most efficacious treatment in dogs of this study. The question of whether or not polyps represent preneoplastic lesions remains unanswered and constitutes an area for future investigation. PMID- 12892300 TI - Prevalence of mucopolysaccharidosis type VI mutations in Siamese cats. AB - Mucopolysaccharidosis type VI (MPS VI), a lysosomal storage disease, is one of the more prevalent inherited diseases in cats and is commonly found in cats with Siamese ancestry. The prevalence of 2 known MPS VI mutations in cats was investigated in 101 clinically normal Siamese cats, in 2 cats with clinical signs of MPS VI, and in 202 cats from 4 research colonies. The mutation L476P which causes a severe clinical phenotype, was present on both alleles in the known MPS VI cats from Italy and North America and was present in all research colonies that originated from North America. However, LA76P was not detected in the Siamese population screened. In contrast, the mutation D520N, which causes a mild clinical phenotype, was identified in 23 of 202 (11.4%) alleles tested in Siamese cats from 3 continents, 2 of which were homozygous for D520N. Thus, the D520N mutation was widespread, and it is likely that cats inheriting both mutations (LA76P/D520N compound heterozygotes) would be in the general Siamese population, particularly in North America. Practitioners should note the high incidence of degenerative joint disease in these animals. PMID- 12892302 TI - Peak mean myocardial velocities and velocity gradients measured by color M-mode tissue Doppler imaging in healthy cats. AB - We sought to assess the feasibility of recording the myocardial velocity gradients (MVGs) and mean myocardial velocities (MMVs) measured by color M-mode tissue Doppler imaging (TDI) in the free wall of unsedated normal cats (n = 18) with a 7.4-MHz probe equipped to record TDI images. The peak MVG and MMV values during the different phases of the cardiac cycle corresponded to certain color velocity patterns occurring in the left ventricular free wall (LVFW). Biphasic shifts were recorded in the tracings of both the MVG and MMV during early diastole (E1 and E2) as well as during the isovolumic relaxation (IVR) and isovolumic contraction (IVC) phases. Stepwise regression analysis showed that age was the only significant predictor for the peak MVG values during the 2nd phase of early diastole (E2) (r = -0.79, r2 = 0.63, and P < .001). The peak late diastolic MVG values were associated positively with age (r = 0.50, r2 = 0.25, and P < .05). The peak MMV values showed a negative association with age during E2 (r = -0.71, r2 = 0.50, and P < .001) as well as during early systole (Se) (r = -0.55, r2 = 0.30, and P < .05) and late systole (SI) (r = -0.62, r2 = 0.39, and P < .01). A positive association was found between age and the peak MMV values during late diastole (r = 0.54, r2 =- 0.29, and P < .05). The MVG values showed cyclic variations consistent with wall thickness changes. The accuracy of velocity determination and the spatial resolution of the system used were validated with a phantom. To our knowledge, this study is the 1st report of the application of this technique to the myocardium of cats,providing insights into the physiology of myocardial motion. It provides reference ranges of the peak MVG and MMV values for future studies of feline myocardial diseases. PMID- 12892303 TI - Use of pericardial fluid pH to distinguish between idiopathic and neoplastic effusions. AB - Pericardial effusion (PE) resulting from neoplasia usually is associated with a poor prognosis, whereas idiopathic PE frequently has a good prognosis. This study examined the utility of pH measurement to distinguish between these 2 etiologies. Dogs were classified as having idiopathic PE (n = 12) if pericarditis was diagnosed on histopathology (n = 4) or if no historical, physical, or echocardiographic evidence of recurrent PE was present for at least 6 months after pericardiocentesis (n = 8). Dogs were classified as having neoplastic PE (n = 25) if pericardial or myocardial neoplasia was detected on histopathology (n = 11) or a discrete mass associated with the right atrium, right ventricle, or the aorta was visualized on echocardiography (n = 14). Samples of PE were centrifuged and the supernatant pH was measured with a portable pH meter. The lowest pH (6.40) was found in a dog with idiopathic PE and the highest pH (7.85) was found in a dog with neoplastic PE. However, data from the 2 groups overlapped in 33 out of 37 (89%) instances, and median pH from the idiopathic and neoplastic groups was not significantly different (7.40 and 7.47, respectively; P = 0.28; difference in medians = -0.7; 95% CI, -0.26-0.06). Because of the degree of overlap, our data provide little justification for the use of pH measurement as a diagnostic test in cases of PE. PMID- 12892304 TI - Thoracolumbar spinal cord compression due to vertebral process degenerative joint disease in a family of Shiloh Shepherd dogs. AB - Five young Shiloh Shepherd Dogs (4 males and 1 female) related by a common sire were studied because of progressive pelvic limb weakness and incoordination. All dogs had a spastic paraparesis and pelvic limb ataxia consistent with an upper motor neuron and general proprioceptive lesion between spinal cord segments T3 and L3. Proliferative lesions involving one or more of the articular processes from the 11th thoracic vertebrae to the 2nd lumbar vertebra were observed on radiographs of the thoracolumbar vertebrae. Dorsal compression of the spinal cord was identified during imaging studies at these sites. Abnormalities of the synovial joints and bony proliferation of the involved articular processes were identified at postmortem examination in 2 dogs. The articular processes and associated vertebral arches protruded into the vertebral canal, indenting the dorsal surface of the spinalcord. Degenerative joint disease (DJD) was identified histologically. A compressive myelopathy was diagnosed in the spinal cord. These dogs were affected by a compressive myelopathy as a consequence of vertebral process DJD that likely has a geneticcomponent. The DJD could have been caused by a primary vertebral malformation or an injury to the processes at a young age causing malarticulation. PMID- 12892305 TI - Polymerase chain reaction (PCR) amplification of parvoviral DNA from the brains of dogs and cats with cerebellar hypoplasia. AB - Cerebellar hypoplasia in cats is caused most commonly by an in utero or perinatal infection with feline panleukopenia virus (parvovirus). Cerebellar hypoplasia has been reported infrequently in dogs, but no viral etiology has been identified to date. DNA was extracted from archival, paraffin-embedded, cerebellar tissue from 8 cats and from 2 canine littermates with cerebellar hypoplasia, 2 canine littermates with cerebellar cortical abiotrophy, 6 dogs with congenital cerebellar vermal defects, 1 dog with congenital hydranencephaly, and 15 dogs and cats with various encephalitdes. The DNA extracted from each cerebellum was subject to polymerase chain reaction (PCR) amplification by 3 primer pairs specific for parvovirus DNA. Sequence analysis of PCR products from each of the 8 cats and 2 dogs with cerebellar hypoplasia confirmed their identity with parvoviral DNA. The 6 dogs with cerebellar vermal defects, 2 dogs with cortical abiotrophy, 1 dog with congenital hydranencephaly, and all control samples were PCR negative for parvovirus. Parvoviral structural proteins were not identified by immunohistochemistry in either dog with cerebellar hypoplasia. This study shows that parvoviral DNA can be amplified from feline and canine archival brain tissue and that cerebellar hypoplasia in dogs might be associated with in utero parvovirus infection. PMID- 12892306 TI - Evaluation of jitter by stimulated single-fiber electromyography in normal dogs. AB - Single-fiber electromyography (SFEMG), a technique used to investigate neuromuscular transmission, has been described previously in the pelvic limb of dogs. Because preferential involvement of isolated muscle groups can occur in disorders of neuromuscular transmission, SFEMG was done in the peroneus longus (PL), extensor carpi radialis (ECR), and orbicularis oculi (OO) muscles of 10 adult, clinically normal dogs. Jitter was calculated as the mean absolute value of the consecutive differences in latency of 50 single muscle fiber action potentials after stimulation of intramuscular nerve bundles at the level of the motor point in at least 20 muscle fibers per muscle. Bilateral recordings were performed in 3 dogs. Mean jitter values were determined for each muscle, and differences among muscle groups and among dogs were compared. The upper limits of mean consecutive difference (mean plus 3 standard deviations) for the PL, ECR, and OO muscles were 21.94, 22.53, and 23.39 micros, respectively, and the upper limit of mean consecutive difference for individual muscle fibers in the respective fiber pools was 28.62, 36.39, and 35.68 micros. Jitter values for the ECR and OO were significantly higher than the jitter value for the PL muscle (P < .05). Significant differences among muscles or dogs or between sides were not observed for the ECR. Significant differences among dogs were observed for OO jitter values and were attributed to extremely low jitter values in 1 dog. Significant differences were demonstrated between sides for the PL and were attributed to small sample size. Results of this study provide normative data that can be used in the application of the stimulated SFEMG technique to dogs with suspected disorders of neuromuscular transmission. PMID- 12892307 TI - L-2-Hydroxyglutaric aciduria in Staffordshire Bull Terriers. AB - L-2-Hydroxyglutaric aciduria is an inborn error of metabolism, which has been recognized in humans since 1980. The metabolic defect responsible for the disease is unknown, but the disorder can be diagnosed in humans by elevations of the organic acid, L-2-hydroxyglutaric acid in the cerebrospinal fluid (CSF), plasma, and urine of affected patients. The disorder produces a variety of clinical neurological defects in humans including psychomotor retardation, seizures, and ataxia. There have previously been no recognized animal models of the disease. However, 6 Staffordshire Bull Terriers were recently identified with the disorder. The animals presented with a variety of clinical signs, most notably seizures, ataxia, dementia, and tremors. They were all screened for organic acid abnormalities in urine, and CSF and plasma (when available). Levels of L-2 hydroxyglutaric acid were elevated in all body fluids evaluated. The clinical, clinicopathologic, and magnetic resonance imaging (MRI) characteristics associated with L-2-hydroxyglutaric acid in Stafforshire Bull Terriers is reported herein and represents the first veterinary model of this inborn error of metabolism. PMID- 12892308 TI - Environmental and lifestyle risk factors for oral squamous cell carcinoma in domestic cats. AB - Oral squamous cell carcinoma (SCC) is a common malignancy in cats, but little currently is known about its etiology. We examined the relationship between risk of oral SCC and factors such as environmental tobacco smoke, flea control products, and diet in 36 domestic cats with histologically confirmed oral SCC and 112 renal disease control cats presented to a large veterinary referral hospital between 1994 and 2000. Questionnaires were mailed to owners of all study and control cats to assess demographic characteristics, lifestyle factors, and level of chemical exposures 2 years before diagnosis. Multivariate relative risks (RR) were used to estimate the relationships between the various factors and the risk of oral SCC. Flea control product use and diet were significantly associated with risk of oral SCC. Cats that wore a flea collar had 5 times the risk of oral SCC as nonusers, after adjustment for other factors (RR = 5.3; P = .002). In contrast, use of flea shampoo substantially reduced risk. Compared to cats eating mostly dry food, those with high canned food intake had a 3-fold increase in risk (RR = 3.6; P = .014); canned tuna fish intake was independently associated with risk (RR = 4.7; P = .004). Exposure to household environmental tobacco smoke was associated with a nonsignificant 2-fold increase in risk (P = .11). Results of this study suggest that flea control products, diet, and perhaps environmental tobacco smoke might be associated with risk of oral SCC and indicate that further investigation into these relationships is warranted. PMID- 12892310 TI - Bile acids in gastric fluids from llamas and alpacas with and without ulcers. AB - To investigate the relationship between duodenogastric reflux and ulceration, gastric fluid was collected from the 1st (n = 26) and 3rd (n = 45) gastric compartments of llamas and alpacas during postmortem examination. Gastric fluids were analyzed for the presence of bile acids. Additionally, the 3rd compartment was examined grossly for the presence of erosions or ulcers, and the intestines were examined for evidence of fluid distention. Detectable bile acids were found in the 3rd compartment fluid of 1 of 28 camelids without ulcers, 7 of 14 camelids with ulcers, and 3 of 3 camelids with intestinal distention. Third compartment bile content was significantly higher in camelids with ulcers (median = 2.9 micromol/L) or intestinal distention (median = 371.5 micromol/L) than those with neither (median = 0 micromol/L). Bile acids were detected in the 1st gastric compartment in only 2 camelids. These findings suggest a reflux of duodenal contents, including bile acids, into the 3rd compartment in camelids with ulcers, similar to what is seen in camelids with poor intestinal emptying. Whether bile reflux was the cause of the ulcers or occurred after or concurrent to their development remains unknown. PMID- 12892309 TI - Effects of topical perineural capsaicin in a reversible model of equine foot lameness. AB - Capsaicin is a local substance P depleter with dramatic analgesic effects. We tested the hypothesis that the topical application of capsaicin ointment over the palmar digital nerves would attenuate the clinical effects of a reversible model of equine foot lameness. Seven healthy adult horses shod unilaterally with adjustable heart bar shoes were studied in a crossover design for 2 weeks. Grade 5.0/5.0 lameness (nonweight bearing) was induced by tightening the adjustable heart bar shoe. One hour later, capsaicin ointment was applied over the medial and lateral palmar digital nerves 3 cm proximal to the coronary band, or horses were left untreated. One week later, treatment assignments were reversed, and the experiment was repeated. The heart rate was markedly lower in treated than in untreated trials at 20 and 40 minutes after capsaicin and between 1.6 and 3.6 hours after capsaicin (P < .05). The lameness score was markedly decreased in capsaicin-treated horses at 40 minutes and from 1.3 to 4 hours after treatment (P < .05). We conclude that the topical application of capsaicin ointment over the palmar digital nerves provided measurable pain relief for up to 4 hours after treatment (P < .05). The clinical application of this analgesic technique in horses with spontaneous clinical or induced laminitis or other sources of foot pain remains to be shown. PMID- 12892311 TI - Myoelectric activity in the intestine of cows with strangulating obstruction of the distal small intestine. AB - Myoelectric activity in 2 cows instrumented with permanent electrodes in the ileum, cecum, proximal loop of the ascending colon (PLAC), and spiral colon was analyzed after an obstruction developed in the distal small intestine. Results were compared with patterns from a group of 7 normal cows. Myoelectric activity in the ileum immediately orad to the occlusion was characterized by abolition of the migrating myoelectric complex (MMC) and a constant pattern of strong spike bursts of long duration. Cyclic activity was present in all parts of the large intestine, and propagation of phase III activity was evident from proximal to distal. A slight degree of disorganization in phase III propagation was restricted to the spiral colon. Activity cycles tended to be shorter in the cecum and PLAC of both cows with colic than in normal cows, and the intensity of spiking activity was generally lower. Changes in duration of the MMC in the spiral colon (bovine colonic MMC, bcMMC) were inconsistent, but the intensity of spiking activity tended to be lower in phases I and II of both cows compared to controls. The organization of phase III in several spindles typical of the bovine spiral colon was not disrupted, but phase IV of the bcMMC occurred only infrequently. Organized cyclic activity occurred in the large intestine of both cows despite complete disruption of the small intestinal MMC, indicating the presence of mechanisms able to initiate and regulate coordinated myoelectric patterns in the large intestine independent of the small intestine. PMID- 12892312 TI - Parachute left atrioventricular valve causing stenosis and regurgitation in a Thoroughbred foal. PMID- 12892313 TI - Suppurative cholangiohepatitis and enteritis in adult horses. PMID- 12892314 TI - Cardiac amyloidosis in a horse. PMID- 12892315 TI - Plasma thyroid hormone concentrations in dogs competing in a long-distance sled dog race. AB - Plasma thyroxine (T4), 3,5,3'-triiodothyronine (T3), total protein, and albumin concentrations were measured in 15 dogs both before and after completion, and in an additional 16 dogs before and 24 dogs after completion, of a long-distance sled dog race. The plasma T4 concentration (mean +/- SD) decreased significantly from 18.2 +/- 5.4 nmol/L before to 14.3 +/- 3.5 nmol/L after the race in dogs evaluated at both times and decreased significantly from 21.8 +/- 10.5 nmol/L before to 15.8 +/- 4.9 nmol/L after the race in dogs sampled only before or only after the race. The mean plasma T3 concentrations in dogs measured twice decreased significantly from 1.20 +/- 0.48 nmol/L before to 0.74 +/- 0.42 nmol/L after the race, as well as in dogs measured either before (1.28 +/- 0.36 nmol/L) or after (0.69 +/- 0.28 nmol/L) the race, respectively. Plasma total protein and albumin concentrations decreased significantly after completion of the race. No significant change was noted in 4 control dogs that did not compete in the race and were tested during a similar time period. The plasma concentrations of T4 and T3 were lower than the normal reference range established for this laboratory in 23 and 39%, respectively, of Alaskan sled dogs tested before the race. Plasma thyroid hormone concentrations frequently are below normal in conditioned Alaskan sled dogs and are further reduced after prolonged submaximal exercise. PMID- 12892316 TI - Cardiac troponin I in pastured and race-training Thoroughbred horses. AB - Cardiac troponin I (cTnI), a myocardial polypeptide, is a highly sensitive and specific biomarker of myocardial injury in people and dogs. The structure of cTnI is highly conserved across species, and equine myocardium has high reactivity with human immunoassays. The purpose of this study was to describe cTnI concentrations in normal pastured and race-training Thoroughbred horses. Ten horses on pasture and 10 horses in race training were studied. Horses were considered normal on the basis of physical examination, training performance, electrocardiography (ECG), and echocardiography. Serum cTnI concentrations were determined with a colorimetric immunoassay. The assay has an analytical sensitivity of 0.04 ng/mL. Serum cTnI concentrations in race-training horses were not significantly different from those of pastured horses. When groups were combined, mean cTnI concentration (+/- SD) was 0.047 +/- 0.085 ng/mL. and the median was 0 (range, 0-0.35 ng/mL). The 90th percentile for both groups combined was 0.11 ng/mL. This study establishes a preliminary reference range for serum cTnI in normal Thoroughbred horses. PMID- 12892317 TI - Dipeptidyl-peptidase IV from bench to bedside: an update on structural properties, functions, and clinical aspects of the enzyme DPP IV. AB - Dipeptidyl-peptidase IV/CD26 (DPP IV) is a cell-surface protease belonging to the prolyloligopeptidase family. It selectively removes the N-terminal dipeptide from peptides with proline or alanine in the second position. Apart from its catalytic activity, it interacts with several proteins, for instance, adenosine deaminase, the HIV gp120 protein, fibronectin, collagen, the chemokine receptor CXCR4, and the tyrosine phosphatase CD45. DPP IV is expressed on a specific set of T lymphocytes, where it is up-regulated after activation. It is also expressed in a variety of tissues, primarily on endothelial and epithelial cells. A soluble form is present in plasma and other body fluids. DPP IV has been proposed as a diagnostic or prognostic marker for various tumors, hematological malignancies, immunological, inflammatory, psychoneuroendocrine disorders, and viral infections. DPP IV truncates many bioactive peptides of medical importance. It plays a role in glucose homeostasis through proteolytic inactivation of the incretins. DPP IV inhibitors improve glucose tolerance and pancreatic islet cell function in animal models of type 2 diabetes and in diabetic patients. The role of DPP IV/ CD26 within the immune system is a combination of its exopeptidase activity and its interactions with different molecules. This enables DPP IV/CD26 to serve as a co-stimulatory molecule to influence T cell activity and to modulate chemotaxis. DPP IV is also implicated in HIV-1 entry, malignant transformation, and tumor invasion. PMID- 12892318 TI - Pseudohyperaldosteronism: pathogenetic mechanisms. AB - Pseudohyperaldosteronism is characterized by a clinical picture of hyperaldosteronism with suppression of plasma renin activity and aldosterone. Pseudohyperaldosteronism can be due to a direct mineralocorticoid effect, as with desoxycorticosterone, fluorohydrocortisone, fluoroprednisolone, estrogens, and the ingestion of high amounts of glycyrrhetinic acid. A block of 11 hydroxysteroid-dehydrogenase type 2 (11HSD2), the enzyme that converts cortisol into cortisone, at the level of epithelial target tissues of aldosterone, is involved in other cases. This mechanism is related either to a mutation of the gene, which encodes 11HSD2 (apparent mineralocorticoid excess syndrome and some cases of low renin hypertension) or to an acquired reduction of the activity of the enzyme due to glycyrrhetinic acid, carbenoxolone, and grapefruit juice. In other cases saturation of 11HSD2 may be involved as in severe Cushing's syndrome and chronic therapy with some corticosteroids. Recently, an activating mutation of the mineralocorticoid receptor gene has been described. Another genetic cause of pseudohyperaldosteronism is the syndrome of Liddle, which is due to a mutation of the gene encoding for beta and gamma subunits of the sodium channels. PMID- 12892319 TI - Calcium and cardiac arrhythmias: DADs, EADs, and alternans. AB - Rapid progress has been made in understanding the molecular mechanisms by which calcium ions mediate certain cardiac arrhythmias. Principal advances include imaging of cytosolic calcium in isolated cells and in intact tissues, use of fluorescent indicators and monophasic action potentials to record membrane potentials in isolated tissue, and sequencing of the genes that encode critical ion channel proteins. In this review, five types of arrhythmias are discussed where calcium ion currents, or currents controlled by calcium, appear to be responsible for arrythmogenesis. These include: (1) the delayed afterpotential that occurs in conditions of intracellular calcium overload such as digitalis toxicity; (2) the early afterdepolarization that occurs when action potential duration is prolonged; (3) the slowly conducted calcium-dependent action potential (the slow response) in the SA and AV nodes; (4) the phenomenon of calcium transient alternans during ischemia, which is related to action potential duration alternans and t-wave alternans; (5) catecholamine-induced cardiac arrhythmias in families with mutations of the sarcoplasmic reticulum calcium release channel. For each type of arrhythmia, the clinical implications of emerging knowledge are discussed. An especially important issue is whether ventricular fibrillation during acute coronary artery occlusion is due to calcium transient alternans. Ventricular fibrillation due to acute ischemia is an important subset of the 400,000 sudden cardiac deaths that occur annually in the U.S. Certain drugs, including beta blockers, fish oils, verapamil, and diltiazem, seem to specifically prevent ventricular fibrillation in this setting, and in most cases an effect of the drug on cytosolic calicum appears to be involved. PMID- 12892320 TI - Electrode systems for measuring cardiac impedances using optical transmembrane potential sensors and interstitial electrodes--theoretical design. AB - The cardiac electrical substrate is a challenge to direct measurement of its properties. Optical technology together with the capability to fabricate small electrodes at close spacings opens new possibilities. Here, those possibilities are explored from a theoretical viewpoint. It appears that with careful measurements from a well-designed set of electrodes one can obtain structural conductivities, separating intracellular from interstitial values, and longitudinal from transverse. Resting membrane resistance also can be obtained. PMID- 12892321 TI - A resampling method for estimating the signal subspace of spatio-temporal EEG/MEG data. AB - Source localization using spatio-temporal electroencephalography (EEG) and magnetoencephalography (MEG) data is usually performed by means of signal subspace methods. The first step of these methods is the estimation of a set of vectors that spans a subspace containing as well as possible the signal of interest. This estimation is usually performed by means of a singular value decomposition (SVD) of the data matrix: The rank of the signal subspace (denoted by r) is estimated from a plot in which the singular values are plotted against their rank order, and the signal subspace itself is estimated by the first r singular vectors. The main problem with this method is that it is strongly affected by spatial covariance in the noise. Therefore, two methods are proposed that are much less affected by this spatial covariance, and old and a new method. The old method involves prewhitening of the data matrix, making use of an estimate of the spatial noise covariance matrix. The new method is based on the matrix product of two average data matrices, resulting from a random partition of a set of stochastically independent replications of the spatio-temporal data matrix. The estimated signal subspace is obtained by first filtering out the asymmetric and negative definite components of this matrix product and then retaining the eigenvectors that correspond to the r largest eigenvalues of this filtered data matrix. The main advantages of the partition-based eigen decomposition over prewhited SVD is that 1) it does not require an estimate of the spatial noise covariance matrix and 2b) that it allows one to make use of a resampling distribution (the so-called partitioning distribution) as a natural quantification of the uncertainty in the estimated rank. The performance of three methods (SVD with and without prewhitening, and the partition-based method) is compared in a simulation study. From this study, it could be concluded that prewhited SVD and the partition-based eigen decomposition perform equally well when the amplitude time series are constant, but that the partition-based method performs better when the amplitude time series are variable. PMID- 12892322 TI - Robust estimation of fetal heart rate variability using Doppler ultrasound. AB - This paper presents a new measure of heart rate variability (HRV) that can be estimated using Doppler ultrasound techniques and is robust to variations in the angle of incidence of the ultrasound beam and the measurement noise. This measure employs the multiple signal characterization (MUSIC) algorithm which is a high resolution method for estimating the frequencies of sinusoidal signals embedded in white noise from short-duration measurements. We show that the product of the square-root of the estimated signal-to-noise ratio (SNR) and the mean-square error of the frequency estimates is independent of the noise level in the signal. Since varying angles of incidence effectively changes the input SNR, this measure of HRV is robust to the input noise as well as the angle of incidence. This paper includes the results of analyzing synthetic and real Doppler ultrasound data that demonstrates the usefulness of the new measure in HRV analysis. PMID- 12892323 TI - Differences between pelvic skin and bone landmark identification in different seated positions on spinal-cord injured subjects. AB - The purpose of this paper was to determine the differences between internal and external pelvic landmark locations in different seating positions. A computer tool developed for the registration of two series of images was used to obtain the internal geometry. First, images of the pelvis were acquired by magnetic resonance imaging (MRI) for each subject, in a supine position; internal landmarks were then identified on the images. Second, ultrasound images of the iliac crests were acquired in four seated positions. A registration algorithm was applied to obtain the transformation matrix between the two image reference systems. The MRI anatomical landmarks were, therefore, transferred into the ultrasound referential, to obtain their three-dimensional (3-D) location in the different seating positions. The external landmarks in those seated positions were identified with a 3-D digitizer. The results revealed that generally the internal and external coordinates of corresponding landmarks are statistically different. The differences are not only due to soft tissue thickness but also to different interpretations of the landmarks' locations between the supine and the seated postures. However, these differences generally did not affect significantly the accuracy with which orientation indexes can be estimated (pelvic tilt, obliquity, transverse rotation). Correlations were found between the internal and external coordinates, implying that linear regressions can be established. PMID- 12892324 TI - Ex vivo assessment of trabecular bone structure from three-dimensional projection reconstruction MR micro-images. AB - Magnetic resonance (MR) imaging has recently been proposed for assessing osteoporosis and predicting fracture risks. However, accurate acquisition techniques and image analysis protocols for the determination of the trabecular bone structure are yet to be defined. The aim of this study was to assess the potential of projection reconstruction (PR) MR microscopy in the analysis of the three-dimensional (3-D) architecture of trabecular bone and in the prediction of its biomechanical properties. High-resolution 3-D PR images (41 x 41 x 82 microm3 voxels) of 15 porcine trabecular bone explants were analyzed to determine the trabecular bone volume fraction (Vv), the mean trabecular thickness (Tb.Th), and the mean trabecular separation (Tb.Sp) using the method of directed secants. These parameters were then compared with those derived from 3-D conventional spin echo microimages. In both cases, segmentation of the high-resolution images into bone and bone marrow was obtained using a spatial adaptive threshold. The contemporary inclusion of Vv, Tb.Th and 1/Tb.Sp in a multiple regression analysis significantly improved the prediction of Young's modulus (YM). The parameters derived from the PR spin-echo images were found to be stronger predictors of YM (R2 = 0.94, p = 0.004) than those derived from conventional spin-echo images (R2 = 0.79, p = 0.051). Our study indicates that projection reconstruction MR microscopy appears to be more accurate than the conventional Fourier transform method in the quantification of trabecular bone structure and in the prediction of its bioimechanical properties. The proposed PR approach should be readily adaptable to the in vivo MRI studies of osteoporosis. PMID- 12892325 TI - Computer-aided method for quantification of cartilage thickness and volume changes using MRI: validation study using a synthetic model. AB - The primary objective of this study was to develop a computer-aided method for the quantification of three-dimensional (3-D) cartilage changes over time in knees with osteoarthritis (OA). We introduced a local coordinate system (LCS) for the femoral and tibial cartilage boundaries that provides a standardized representation of cartilage geometry, thickness, and volume. The LCS can be registered in different data sets from the same patient so that results can be directly compared. Cartilage boundaries are segmented from 3-D magnetic resonance (MR) slices with a semi-automated method and transformed into offset-maps, defined by the LCS. Volumes and thickness are computed from these offset-maps. Further anatomical labeling allows focal volumes to be evaluated in predefined subregions. The accuracy of the automated behavior of the method was assessed, without any human intervention, using realistic, synthetic 3-D MR images of a human knee. The error in thickness evaluation is lower than 0.12 mm for the tibia and femur. Cartilage volumes in anatomical subregions show a coefficient of variation ranging from 0.11% to 0.32%. This method improves noninvasive 3-D analysis of cartilage thickness and volume and is well suited for in vivo follow up clinical studies of OA knees. PMID- 12892326 TI - Assessment of the 3-d reconstruction and high-resolution geometrical modeling of the human skeletal trunk from 2-D radiographic images. AB - This paper presents an in vivo validation of a method for the three-dimensional (3-D) high-resolution modeling of the human spine, rib cage, and pelvis for the study of spinal deformities. The method uses an adaptation of a standard close range photogrammetry method called direct linear transformation to reconstruct the 3-D coordinates of anatomical landmarks from three radiographic images of the subject's trunk. It then deforms in 3-D 1-mm-resolution anatomical primitives (reference bones) obtained by serial computed tomography-scan reconstruction of a dry specimen. The free-form deformation is calculated using dual kriging equations. In vivo validation of this method on 40 scoliotic vertebrae gives an overall accuracy of 3.3 +/- 3.8 mm, making it an adequate tool for clinical studies and mechanical analysis purposes. PMID- 12892327 TI - A wavelet-based method for action potential detection from extracellular neural signal recording with low signal-to-noise ratio. AB - We present a method for the detection of action potentials, an essential first step in the analysis of extracellular neural signals. The low signal-to-noise ratio (SNR) and similarity of spectral characteristic between the target signal and background noise are obstacles to solving this problem and, thus, in previous studies on experimental neurophysiology, only action potentials with sufficiently large amplitude have been detected and analyzed. In order to lower the level of SNR required for successful detection, we propose an action potential detector based on a prudent combination of wavelet coefficients of multiple scales and demonstrate its performance for neural signal recording with varying degrees of similarity between signal and noise. The experimental data include recordings from the rat somatosensory cortex, the giant medial nerve of crayfish, and the cutaneous nerve of bullfrog. The proposed method was tested for various SNR values and degrees of spectral similarity. The method was superior to the Teager energy operator and even comparable to or better than the optimal linear detector. A detection ratio higher than 80% at a false alarm ratio lower than 10% was achieved, under an SNR of 2.35 for the rat cortex data where the spectral similarity was very high. PMID- 12892328 TI - Characterization and imaging of compositional variation in tissues. AB - The diffuse surface reflectance profiles of the goat's isolated heart, spleen, and adipose tissues by multiprobe laser reflectometer are measured. The normalized backscattered intensity values for adipose, heart, and spleen tissues at source-detector separation 0.2 cm, are 0.060, 0.021, and 0.003, respectively. The optical parameters of these tissues are determined by the best fit (chi2(0.99)) of their spatial profiles with that as obtained by Monte Carlo simulation by iterative procedure. As the optical parameters of these vary over a wide range, adipose and spleen tissues are treated as inhomogeneity of diameter 0.1, 0.2, or 0.3 cm, and placed inside the control (heart) tissue at different depths. Anisotropic simulation of light backscattering or photon depth distribution is significantly different for various tissues. The surface intensity profiles vary depending on the changes in tissue composition. From the horizontal scans of the subtracted images, the photon backscattering simulated images of control and combination of tissues are obtained. By analysis of peak intensity and full-width at half maximum, the type, location, and size of the tissue compositional variation are determined. PMID- 12892329 TI - Differentiation among basal cell carcinoma, benign lesions, and normal skin using electric impedance. AB - This paper presents a preliminary study showing the diagnostic potential of electrical impedance to detect basal cell carcinoma (BCC). Electrical impedance was measured in vivo from 1 kHz to 1 MHz on 24 human subjects over BCC (19 lesions), over benign tumors (11 lesions), and over normal skin (all 24 patients). Lesions ranged from 2-15 mm in diameter. Indexes based on the magnitude (MIX), phase (PIX), real-part (RIX) and imaginary-part (IMIX) of impedance were calculated for each measurement. Significant differences were found between measurements over BCC, benign lesions and normal skin for indexes MIX, PIX, and IMIX (P = 0.04 to P = 7 x 10(-7)). Indexes were generally smaller for measurements of BCC than for benign lesions or normal skin. Differences were not a result of differences in the patient's age or the measurement location. The large size of our measurement electrode (10 mm) probably limited our ability to differentiate lesions because significant amounts of normal skin were included in each lesion measurement. A linear regression fit of data with tumor size suggests that a smaller probe or more sophisticated analysis techniques may improve differentiation. Results suggest that electrical impedance could be used to provide rapid and noninvasive differentiation of BCC from similar looking benign lesions. PMID- 12892330 TI - Optical transmission of blood: effect of erythrocyte aggregation. AB - The influence of red blood cell (RBC) aggregation on transparency of blood in the red-near infrared spectral range is investigated. We argue that for relatively thin blood layers the light diffraction on aggregates becomes the dominant phenomenon. The nature of pulsatile changes of blood transparency is explained by pulsations of RBC aggregate size. For another case of over-systolic vessel occlusion the following time evolution of blood transparency strongly depends on light wavelength. This dependence may serve as a basis for an alternative approach to noninvasive blood tests: occlusion spectroscopy. Theoretical results well correspond to both in vivo and in vitro measurements reproducing pulsatile blood flow and long occlusion as well. PMID- 12892331 TI - Comments on "an efficient coding algorithm for the compression of ECG signals using the wavelet transform". AB - The author proposed an effective wavelet-based ECG compression algorithm (Rajoub, 2002). The reported extraordinary performance motivated us to explore the findings and to use it in our research activity. During the implementation of the proposed algorithm several important points regarding accuracy, methodology, and coding were found to be improperly substantiated. This paper discusses these findings and provides specific subjective and objective measures that could improve the interpretation of compression results in these research-type problems. PMID- 12892332 TI - Myocardial kinematics from tagged MRI based on a 4-D B-spline model. AB - Current research investigating the modeling of left ventricular dynamics for accurate clinical assessment of cardiac function is extensive. Magnetic resonance (MR) tagging is a functional imaging method which allows for encoding of a grid of signal voids on cardiac MR images, providing a mechanism for noninvasive measurement of intramural tissue deformations, in vivo. We present a novel technique of employing a four-dimensional (4-D) B-spline model which permits concurrent determination of myocardial beads and myocardial strains. The method entails fitting the knot planes of the 4-D B-spline model for fixed times to a sequence of triplets of orthogonal sets of tag surfaces for all imaged volumetric frames within the constraints of the model's spatio-temporal internal energy. From a three-dimensional (3-D) displacement field, the corresponding long and short-axis Lagrangian normal, shear, and principal strain maps are produced. As an important byproduct, the points defined by the 3-D intersections of the triplets of orthogonal tag planes, which we refer to as myocardial beads, can easily be determined by our model. Displaying the beads as a movie loop allows for the visualization of the nonrigid movement of the left ventricle in 3-D. PMID- 12892333 TI - Logic and teleologic. PMID- 12892334 TI - The Air Force Dental Service--cross into the blue. AB - The Air Force Dental Service was founded shortly after World War II in order to provide oral health care to active duty, retired, and active reserve Air Force personnel, their families while overseas, and designated others. Grouped with medical facilities and organized by functional commands, the Air Force Dental Service enjoys a strong reputation in special operations, including education, forensics, expeditionary services (including combat and humanitarian efforts), the Dental Investigation Service, and the Air Force Inspection Agency. PMID- 12892335 TI - Providing care for America's Army. AB - The Army Dental Corps' three-part mission is to maintain soldiers fit for combat, promote health, and ensure the Dental Corps ability deploy and deliver in the field. Consistent with this mission, the corps is developing innovative dental delivery systems and promoting tobacco cessation, sealants, mouth guard use, cancer detection, and identification of child, elder, and other abuse. The corps' training programs include options and benefits at the dental student, postdoctoral residency, and specialty levels. Recent technology innovations include light-weight field equipment, an integrated computer database to manage treatment, rapid ordering and delivery of supplies, and distance education. PMID- 12892336 TI - Navy Dental Corps: ninety years ... and forward. AB - The Navy Dental Corps is responsible for ensuring the readiness of America's sailors and marines and optimizing their oral health. This article traces the history from the 1912 Act of Congress authorizing thirty "assistant dental surgeons" as the first Navy Dental Corps through service around the world. Navy dentists have seen service in every war and action in the past ninety years, reaching a peak of seven thousand officers and eleven thousand technicians in World War II. The Navy Dental Corps has served in the Korean and Vietnam Wars, Beirut, Somalia, Haiti, 9/11, Desert Storm, Desert Shield, and Operation Iraqi Freedom. PMID- 12892337 TI - Promoting the public's oral health: the Department of Health and Human Services, U.S. Public Health Service, and the U.S. Public Health Service Commissioned Corps. AB - The story of the Department of Health and Human Services (DHHS), its United States Public Health Service (US PHS), and the US PHS Commissioned Corps is comprised of people and programs aimed at protecting and promoting the nation's health, including oral health. The federal precursors of these organizations focused on clinical services for federal beneficiaries, and with time grew to include federal support for community and state programs for underserved and institutionalized populations; biomedical and behavioral research conduct: drug, device, and food regulatory activities; and, most recently, an enhanced response to biodefense and emergency readiness, among other activities. An essential component of the workforce addressing these activities is the US PHS Commissioned Corps, directed by the Surgeon General of the US PHS. This corps is a mobile, uniformed health service assigned to programs throughout the DHHS, as well as to other departments and agencies as needed. Dentistry has been a critical part of these programs and of the corps since their inception. PMID- 12892339 TI - Integrity and compromise in dental ethics. PMID- 12892338 TI - Dentistry in the Department of Veterans Affairs. AB - The dental mission of the Department of Veterans Affairs includes care, typically hospital care, developing an educated cadre of oral health care professionals, and research. Its innovations emphasize the local nature of care requirements, and it enjoys a tradition of partnering with other branches of the federal services, the American Dental Association, and education. PMID- 12892340 TI - Errors. AB - Errors are outcomes or patterns of outcomes you don't want to see. They are basically of two types: those that occur naturally as rare outcomes of effective processes and those that signal that the process is not working. The error rate in American medicine is known to be alarmingly large; it has not been looked at in dentistry. A topology of error is presented and it is shown how understanding the conditions that cause error, and thus the prevention of error, can only be developed by recognizing and analyzing error itself. PMID- 12892341 TI - Sexual history taking should be taught in medical school. PMID- 12892342 TI - Should pelvic examination be continued after hysterectomy? PMID- 12892343 TI - Diagnostic testing for HIV infection. PMID- 12892344 TI - JNC 7 express: new thinking in hypertension treatment. PMID- 12892345 TI - Should we offer routine breast cancer screening with mammography? PMID- 12892346 TI - Management of carpal tunnel syndrome. AB - Carpal tunnel syndrome affects approximately 3 percent of adults in the United States. Pain and paresthesias in the distribution of the median nerve are the classic symptoms. While Tinel's sign and a positive Phalen's maneuver are classic clinical signs of the syndrome, hypalgesia and weak thumb abduction are more predictive of abnormal nerve conduction studies. Conservative treatment options include splinting the wrist in a neutral position and ultrasound therapy. Orally administered corticosteroids can be effective for short-term management (two to four weeks), but local corticosteroid injections may improve symptoms for a longer period. A recent systematic review demonstrated that nonsteroidal anti inflammatory drugs, pyridoxine, and diuretics are no more effective than placebo in relieving the symptoms of carpal tunnel syndrome. If symptoms are refractory to conservative measures or if nerve conduction studies show severe entrapment, open or endoscopic carpal tunnel release may be necessary. Carpal tunnel syndrome should be treated conservatively in pregnant women because spontaneous postpartum resolution is common. PMID- 12892347 TI - Information from your family doctor. Carpal tunnel syndrome. PMID- 12892349 TI - Information from your family doctor. Syphilis. PMID- 12892348 TI - Diagnosis and management of syphilis. AB - Syphilis is a sexually transmitted disease with varied and often subtle clinical manifestations. Primary syphilis typically presents as a solitary, painless chancre, whereas secondary syphilis can have a wide variety of symptoms, especially fever, lymphadenopathy, rash, and genital or perineal condyloma latum. In latent syphilis, all clinical manifestations subside, and infection is apparent only on serologic testing. Late or tertiary syphilis can manifest years after infection as gummatous disease, cardiovascular disease, or central nervous system involvement. Neurosyphilis can develop in any stage of syphilis. The diagnosis of syphilis may involve dark-field microscopy of skin lesions but most often requires screening with a nontreponemal test and confirmation with a treponemal-specific test. Parenterally administered penicillin G is considered first-line therapy for all stages of syphilis. Alternative regimens for nonpregnant patients with no evidence of central nervous system involvement include doxycycline, tetracycline, ceftriaxone, and azithromycin. In pregnant women and patients with neurosyphilis, penicillin remains the only effective treatment option; if these patients are allergic to penicillin, desensitization is required before treatment is initiated. Once the diagnosis of syphilis is confirmed, quantitative nontreponemal test titers should be obtained. These titers should decline fourfold within six months after treatment of primary or secondary syphilis and within 12 to 24 months after treatment of latent or late syphilis. Serial cerebrospinal fluid examinations are necessary to ensure adequate treatment of neurosyphilis. PMID- 12892350 TI - Vaccinations in pregnancy. AB - Adult immunization rates have fallen short of national goals partly because of misconceptions about the safety and benefits of current vaccines. The danger of these misconceptions is magnified during pregnancy, when concerned physicians are hesitant to administer vaccines and patients are reluctant to accept them. Routine vaccines that generally are safe to administer during pregnancy include diphtheria, tetanus, influenza, and hepatitis B. Other vaccines, such as meningococcal and rabies, may be considered. Vaccines that are contraindicated, because of the theoretic risk of fetal transmission, include measles, mumps, and rubella; varicella; and bacille Calmette-Guerin. A number of other vaccines have not yet been adequately studied; therefore, theoretic risks of vaccination must be weighed against the risks of the disease to mother and fetus. Inadvertent administration of any of these vaccinations, however, is not considered an indication for termination of the pregnancy. PMID- 12892351 TI - Necrotizing (malignant) external otitis. AB - Necrotizing (malignant) external otitis, an infection involving the temporal and adjacent bones, is a relatively rare complication of external otitis. It occurs primarily in immunocompromised persons, especially older persons with diabetes mellitus, and is often initiated by self-inflicted or iatrogenic trauma to the external auditory canal. The most frequent pathogen is Pseudomonas aeruginosa. Patients with necrotizing external otitis complain of severe otalgia that worsens at night, and otorrhea. Clinical findings include granulation tissue in the external auditory canal, especially at the bone-cartilage junction. Facial and other cranial nerve palsies indicate a poor prognosis; intracranial complications are the most frequent cause of death. Diagnosis requires culture of ear secretions and pathologic examination of granulation tissue from the infection site. Imaging studies may include computed tomographic scanning, technetium Tc 99m medronate bone scanning, and gallium citrate Ga 67 scintigraphy. Treatment includes correction of immunosuppression (when possible), local treatment of the auditory canal, long-term systemic antibiotic therapy and, in selected patients, surgery. Family physicians and others who provide medical care for immunocompromised patients should be alert to the possibility of necrotizing external otitis in patients who complain of otalgia, particularly if they have diabetes mellitus and external otitis that has been refractory to standard therapy. Susceptible patients should be educated to avoid manipulation of the ear canal (i.e., they should not use cotton swabs to clean their ears) and to minimize exposure of the ear canal to water with a high chloride concentration. Appropriate patients should be referred to an otolaryngologist. PMID- 12892352 TI - Necrotizing soft tissue infections: a primary care review. AB - Patients with necrotizing soft tissue infections often present initially to family physicians. These infections must be detected and treated rapidly to prevent loss of limb or a fatal outcome. Unfortunately, necrotizing soft tissue infections have no pathognomonic signs. Patients may present with some evidence of cellulitis, vesicles, bullae, edema, crepitus, erythema, and fever. They also may complain of pain that seems out of proportion to the physical findings; as the infection progresses, their pain may decrease. Magnetic resonance imaging and laboratory findings such as acidosis, anemia, electrolyte abnormalities, coagulopathy, and an elevated white blood cell count may provide clues to the diagnosis. No single organism or combination of organisms is consistently responsible for necrotizing soft tissue infections. Most infections are polymicrobial, with both anaerobic and aerobic bacteria frequently present. Fungal infections also have been reported. Generally, bacterial and toxin-related effects converge to cause skin necrosis, shock, and multisystem organ failure. Aggressive debridement of infected tissues is critical to management. Antimicrobial therapy is important but remains secondary to the removal of diseased and necrotic tissues. PMID- 12892354 TI - Information from your family doctor. Heart failure. PMID- 12892353 TI - Screening for gestational diabetes mellitus: recommendation and rationale. PMID- 12892355 TI - Information from your family doctor. Heart attack: warning signs and tips on prevention. PMID- 12892356 TI - Information from your family doctor. Heart attack: getting back into your life after a heart attack. PMID- 12892357 TI - NHLBI releases new high blood pressure guidelines. PMID- 12892358 TI - Critical review of non-invasive respiratory monitoring in medical care. AB - Respiratory failure can be difficult to predict. It can develop into a life threatening condition in just a few minutes, or it can build up more slowly. Thus continuous monitoring of respiratory activity should be mandatory in clinical, high-risk situations, and appropriate monitoring equipment could be life-saving. The review considers non-invasive methods and devices claimed to provide information about respiratory rate or depth, or gas exchange. Methods are categorised into those responding to movement, volume and tissue composition detection; air flow; and blood gas concentration. The merits and limitations of the methods and devices are analysed, considering information content and their ability to minimise the rate of false alarms and false non-alarms. It is concluded that the field of non-invasive respiratory monitoring is still in an exploratory phase, with numerous reports on specific device solutions but less work on evaluation and adaptation to clinical requirements. Convincing evidence of the clinical usefulness of respiratory monitors is still lacking. Devices responding only to respiratory rate, and lacking information about actual gas exchange, will have limited clinical value. Furthermore, enhancement in specificity and sensitivity to avoid false alarms and non-alarms will be necessary to meet clinical requirements. Miniature CO2 sensors are identified as one route towards substantial improvement. PMID- 12892359 TI - Bio-engineering inslulin-secreting cells from embryonic stem cells: a review of progress. AB - According to the Edmonton protocol, human islet transplantation can result in insulin independency for periods longer than 3 years. However, this therapy for type 1 diabetes is limited by the scarcity of cadaveric donors. Owing to the ability of embryonic stem cells to expand in vitro and differentiate into a variety of cell types, research has focused on ways to manipulate these cells to overcome this problem. It has been demonstrated that mouse embryonic stem cells can differentiate into insulin-containing cells, restoring normoglycaemia in diabetic mice. To this end, mouse embryonic stem cells were transfected with a DNA construct that provides resistance to neomycin under the control of the regulatory regions of the human insulin gene. However, this protocol has a very low efficiency, needing improvements for this technology to be transferred to human stem cells. Optimum protocols will be instrumental in the production of an unlimited source of cells that synthesise, store and release insulin in a physiological manner. The review focuses on the alternative source of tissue offered by embryonic stem cells for regenerative medicine in diabetes and some key points that should be considered in order for a definitive protocol for in vitro differentiation to be established. PMID- 12892360 TI - Dialysate-side urea kinetics. Neural network predicts dialysis dose during dialysis. AB - Determination of the adequacy of dialysis is a routine but crucial procedure in patient evaluation. The total dialysis dose, expressed as Kt/V, has been widely recognised to be a major determinant of morbidity and mortality in haemodialysed patients. Many different factors influence the correct determination of Kt/V, such as urea sequestration in different body compartments, access and cardiopulmonary recirculation. These factors are responsible for urea rebound after the end of the haemodialysis session, causing poor Kt/V estimation. There are many techniques that try to overcome this problem. Some of them use analysis of blood-side urea samples, and, in recent years, on-line urea monitors have become available to calculate haemodialysis dose from dialysate-side urea kinetics. All these methods require waiting until the end of the session to calculate the Kt/V dose. In this work, a neural network (NN) method is presented for early prediction of the Kt/V dose. Two different portions of the dialysate urea concentration-time profile (provided by an on-line urea monitor) were analysed: the entire curve A and the first half B, using an NN to predict the Kt/V and compare this with that provided by the monitor. The NN was able to predict Kt/V is the middle of the 4h session (B data) without a significant increase in the percentage error (B data: 6.69% +/- 2.46%; A data: 5.58% +/- 8.77%, mean +/- SD) compared with the monitor Kt/V. PMID- 12892361 TI - K-means clustering method for auditory evoked potentials selection. AB - Surface auditory evoked potentials are generally recorded using a headset of 32, 64 or 128 electrodes, but the quality of the responses is quite heterogeneous on the scalp surface. In some contexts, such as the analysis of auditory evoked potentials recorded in radio-frequency fields, the signal quality is essential, and it appears pertinent to consider only a limited number of electrodes. Therefore, before analysing signals influenced by radio-frequency fields, it is necessary to consider the preliminary step of selecting the channels where auditory activity is strong. This step is often realised by human visual selection and can take a considerable time. In this paper, a simple k-means clustering method is proposed, to select automatically the important channels, and the results are compared with traditional methods of selection. The method detected channels that showed a concordance rate of 86.5% with the visual selection (performed by five individuals) and gave the same final selection (only two extra electrodes in the automatic case). Moreover, the time needed for this automatic selection was 100 times less than that for the visual selection, and also human variability was avoided. PMID- 12892362 TI - Changes in tissue optical properties due to radio-frequency ablation of myocardium. AB - The optical properties of pig heart tissue were measured after in vivo ablation therapy had been performed during open-heart surgery. In vitro samples of normal and ablated tissue were subjected to measurements with an optically integrating sphere set-up in the region 470-900 nm. Three independent measurements were made: total transmittance, total reflectance and collimated transmittance, which made it possible to extract the absorption and scattering coefficients and the scattering anisotropy factor g, using an inverse Monte Carlo model. Between 470 and 700 nm, only the reduced scattering coefficient and absorption could be evaluated. The absorption spectra were fitted to known tissue chromophore spectra, so that the concentrations of haemoglobin and myoglobin could be estimated. The reduced scattering coefficient was compared with Mie computations to provide Mie equivalent average radii. Most of the absorption was from myoglobin, whereas haemoglobin absorption was negligible. Metmyoglobin was formed in the ablated tissue, which could yield a spectral signature to distinguish the ablated tissue with a simple optical probe to monitor the ablation therapy. The reduced scattering coefficient increased by, on average, 50% in the ablated tissue, which corresponded to a slight decrease in the Mie equivalent radius. PMID- 12892363 TI - Evaluation of electrocardiogram T-wave dispersion measurement methods. AB - Methods for measuring the T-wave dispersion are difficult to improve, because the exact result is unknown. This study describes a flexible and cheap method for analysing the measurement problems. The analysis consisted of measuring the T wave dispersion of electrocardiograms and vector loops with simulated dispersion. The vector loops were simulated using two primary T-waves that were identical, quasi-monophasic and located in orthogonal leads. The dispersion was the time delay between the primary T-waves. The electrocardiograms were projections of the vector loops on approximations to the orientations of leads in the standard 12 lead ECG. This simulation model was validated by matching simulated ECGs and vector loops with measured ECGs and vector loops drawn from a population of 247 patients with acute myocardial infarction. The analysis of the measurement problems was demonstrated by measuring an approximation to the classical QT dispersion in ECGs with simulated dispersion in the interval 0-100 ms, in steps of 2 ms. This showed that the QT-dispersion was unable to measure the simulated dispersion accurately in the clinically relevant interval of 50-100 ms. The results indicated that the low prognostic accuracy of the QT-dispersion could be due to an inaccurate measurement method. PMID- 12892364 TI - Development and validation of implantable sensors for monitoring function of prosthetic heart valves: in vitro studies. AB - The development of a 'smart' heart valve prosthesis, with the intrinsic ability to monitor thrombus formation, mechanical failure and local haemodynamics and to relay this information externally, would be of significant help to clinicians. The first step towards such a valve is development of the sensors and examination of whether sensor output provides predictive information on function. Custom-made piezo-electric sensors were mounted onto the housing of mechanical valves with various layers of simulated thrombus and bioprosthetic valves with normal and stiffened leaflets. Sensor output was examined using joint time-frequency analysis. Sensors were able to detect leaflet opening and closing with high fidelity for all types of valve. The frequency content of the closing sounds for the mechanical valves contained several peaks between 100 Hz and 10 kHz, whereas closing sounds for the bioprosthetic valve contained energy in a lower frequency range (<1 kHz). A frequency peak of 47 +/- 15 Hz was seen for the normal bioprosthetic valve; this peak increased to 115 +/- 12 Hz for the valve with visibly stiffened leaflets. Total low-frequency (80-3500 Hz) energy content diminished predictably with increasing levels of thrombus for the mechanical valves. Lastly, closing sound intensity correlated well with closing pressure dynamics (dp/dt) (y = 190x - 443; r = 0.90), indicating that the sensors also provide information on haemodynamics. These studies provide initial evidence regarding the use of embedded sensors to detect prosthetic valve function. Efforts to encapsulate these sensors with telemetry into a custom valve are currently underway. PMID- 12892365 TI - Adaptive edge localisation approach for quantitative coronary analysis. AB - Lack of reliability, user dissatisfaction and errors in determining coronary vessel wall characteristics are challenging issues in quantitative coronary analysis (QCA). A new approach is proposed for QCA that tackles these issues. The proposed approach extracts the coronary vessel edges by applying dynamic programming techniques that use human-based decision criteria, adaptive edge detection and feature-based cost minimisation. This approach uses image gradient, image intensity, boundary continuity and adaptive thresholding to gain maximum quality assurance. The validation of this approach was conducted through modelled phantoms and real X-ray angiograms. The results show that the accuracies obtained were 0.0116mm and 0.06mm, respectively, and the precisions were 0.0263mm, and 0.04mm, respectively. The proposed approach is reliable, reproducible and user friendly and provides high precision compared with recently published results. Furthermore, the significance of the proposed approach and its limitations are also discussed. PMID- 12892366 TI - Very high frequency oscillations in the heart rate and blood pressure of heart transplant patients. AB - The authors studied the recently reported very high frequency (VHF) peaks in the heart rate (HR) and blood pressure (BP) power spectra of heart transplant (HT) patients. These VHF peaks appear at frequencies much higher than the respiratory frequency, in addition to the typical low-frequency and high-frequency peaks. Twenty-five recordings obtained from 13 male HT patients (0.5-65 months following surgery) were compared with recordings from 14 normal male subjects. The ECG, continuous BP and respiration were recorded during 45min of supine rest. Eight recordings from HT patients were excluded owing to arrhythmias. Spectral analysis was performed on all other recordings. VHF peaks were found in the spectra of both BP and HR in nine recordings obtained from six HT patients. In some cases, the power in the VHF peaks was markedly higher than that of the high-frequency peak. No VHF peaks were observed in eight recordings obtained from four HT patients or in recording from any of the normal subjects. No correlation was found between the incidence of VHF peaks and time after transplant. It was proved that the VHF peaks were not artifactual, and their significance within the framework of the theory of communication systems is discussed. The presence of those peaks was attributed to vagal denervation. PMID- 12892367 TI - Simplified oscillation method for assessing nasal obstruction non-invasively and under spontaneous ventilation: a pilot study. AB - The clinical application of the current methods of measuring nasal obstruction has been limited by complicated, invasive and stressful procedures that require the full co-operation of the patient. A pilot study is described where a simple way of evaluating nasal obstruction, based on oscillation methods, was investigated. The technique did not disturb spontaneous breathing and required little co-operation and comprehension. Significant differences were obtained when clinically classified normal (5.2 +/- 1.8 cmH2O l(-1) s) and patient (10.6 +/- 5.9 cmH2O l(-1) s) groups were evaluated (p<0.01). A significant reduction (p<0.02) was also observed in impedance results before (8.5 +/- 1.1 cmH2O l(-1) s) and after (5.2 +/- 1.7 cmH2O l(-1) s) clinically successful nasal surgery, closely reflecting the clinical conditions of the subjects. This simple forced oscillation technique showed good potential for future clinical applications in the pre-screening of nasal patients and the evaluation of therapeutic surgery. PMID- 12892368 TI - Discrimination analysis of discontinuous breath sounds using higher-order crossings. AB - The paper evaluates the performance of an automatic discrimination analysis (DA) method used to discriminate efficiently the types of discontinuous breath sound (DBS), i.e. fine crackles (FCs), coarse crackles (CCs) and squawks (SQs); this may lead to more accurate characterisation of the pulmonary acoustical changes due to the related pathology. Based on higher-order crossings (HOCs), the proposed method, HOC-DA, captured the differences in the oscillatory patterns of FCs, CCs and SQs, which are only exposed when higher (> 1) crossings are employed. Prior to HOC-DA, wavelet-based de-noising of DBSs was employed to eliminate the effects of the vesicular sound (background noise) from their oscillatory pattern. The HOC-DA was applied to 157 discontinuous breath sounds corresponding to 16 cases included in three lung sound databases. Results showed that the HOC-DA efficiently separated FCs from CCs, SQs from CCs (both with an accuracy of 100%), and SQs from FCs (accuracy of 80%), with the optimum order ranging from 9 to 11. When compared with other classification tools, the HOC-DA resulted in high discrimination accuracy without involving high computational complexity. Owing to its simplicity, it could be implemented in a real-time context and be used in clinical medicine as a module of an integrated intelligent patient evaluation system. PMID- 12892369 TI - Screen-printed transcutaneous oxygen sensor employing polymer electrolytes. AB - A disposable, transcutaneous oxygen sensor has been designed and implemented using screen-printing technology for all fabrication stages. The sensor incorporates an integral heating element to promote transcutaneous diffusion of blood gases so that a reliable estimation of arterial blood gas concentration can be obtained. The oxygen sensing part of the device consists of a screen-printed Clark cell implemented as electrodes, electrolyte and membrane. A three-electrode configuration is employed with gold working and counter electrodes and a silver/silver chloride reference electrode. Several different polymer electrolyte and membrane materials were evaluated in the construction of the device, and their performances were compared. A fully automated gas testing rig was constructed to enable oxygen levels to be varied under computer control. Cyclic voltammetry and static analysis of the sensors were carried out at different oxygen concentration levels and in various test environments. Linear relationships were established with an averaged sensitivity level of 0.02 microA(mmHg)(-1) and high regression coefficients of 0.99. The prototype covered with a polytetrafluoroethylene membrane gave the experimental result of I (microA) = -0.025PO2 (mmHg) - 0.085. Several factors influenced the performance of the sensors. The investigations have greatly contributed towards an understanding of the suitability of the materials in achieving a viable, low-cost sensor. PMID- 12892370 TI - Assessment of a procedure for detecting minute levels of tooth erosion. AB - Over a period of some years, the components of a system for discerning erosion on children's teeth have been progressively developed, for use in an extensive project seeking correlations between erosion and various perceived risk factors. The aim was the detection of minute levels of erosion, based on mappings of the palatal surfaces of the maxillary central incisors in children. Significant challenges were encountered, the primary problem being the impracticality of placing control marks that would aid the realignment of successive measurements. The paper describes the erosion detection system and initial experiences based on the results of the first 100 subjects measured after 9 months. The procedures detected the occurrence of erosion of 50 microm magnitude on about one-quarter of the teeth over the 9 month period, at a precision estimated to be +/- 15 microm. The occurrence of some anomalous incidents prevented the procedure from being fully automatic, and it necessitated human examination of a graphical diagram derived from the surface matching program, but it was nevertheless superior to current practices of examining impressions or replicas entirely by eye. PMID- 12892371 TI - The use of a 'phantom scalp' to assess the possible direct pickup of mobile phone handset emissions by electroencephalogram electrode leads. AB - A 'Phantom Scalp' was constructed from a conducting foam mat to form a layer under a 62-electrode electroencephalogram cap closely approximating the electrical properties of a human scalp. The mat was placed over a polystyrene manikin head to preserve a correct anatomical arrangement. Electrical signals were recorded with a Global System for Mobile (GSM) communications mobile phone handset in place against the side of the 'head'. Amplitude spectra were compared for three phone conditions: 'off', 'standby' and 'transmit'. At 217 Hz, significant differences were obtained between 'transmit' and the other two conditions, but no significant differences were noted for the physiologically important range 0.5-30 Hz. An anomalous difference was noted for one electrode in the range 30-45 Hz. PMID- 12892372 TI - Non-linear registration for brain images by maximising feature and intensity similarities with a Bayesian framework. AB - The objective of this work was to provide a new, precise registration of the cortical mantle with a non-linear transformation. Image registration is broadly classified into two types, using intensity similarity and feature similarity. Whereas the former approach has merit in global brain matching, the latter provides a fast registration centred on a region of interest. The hybrid registration proposed in this paper was achieved using a Bayesian framework, which consisted of a likelihood model including intensity similarity and a prior model including feature information and a smoothing constraint. In this approach, each voxel was spatially transformed, so that the distance between corresponding features was shortened and also so that the intensity correlation was maximised. The result of the hybrid method clearly showed a good match of global brain (r = 0.930) by including intensity similarity. Moreover, this method compensated for the approximated sulcus of the feature-based method with intensity information, so that the geometric shape and thickness of the sulcus at the feature-defined region was likely to be registered. The accuracy in the feature-defined area was improved by 33.4% and 7.5% compared with feature-based and intensity-based methods, respectively. PMID- 12892373 TI - Correlation between ciliary beat frequency and metachronal wave disorder using image analysis method. AB - Ciliary beating and metachronal waves are fundamental to effective mucociliary transport. The ciliary beat frequencies (CBFs) and metachronal wave directions of multiple cilia beating in culture media were measured simultaneously using digital microscopic images. The degree of synchronisation between ciliary beats was determined by the correlation between ciliary signals at two different locations. The wave propagation directions of cilia were determined from a two dimensional correlation map by a principal axis method. The standard deviation of measured wave directions in a region of interest was defined as a measure of metachronal wave disorder (MWD). Considerable variation was found in the beat frequencies and metachronal wave directions of cilia beating on epithelium. The pooled mean of MWDs was 23.4 +/- 8.8 degrees, and the pooled mean of CBFs was 10.1 +/- 1.9 Hz on 120 cells from five healthy subjects. The means of the MWD and the CBF from subjects were highly correlated (correlation = -0.83). The higher the CBF, the lower the level of the MWD. PMID- 12892374 TI - Measurement device for ankle joint kinematic and dynamic characterisation. AB - The paper describes a measurement device for obtaining the kinematic characterisation and isometric loading of ankle joints under different working conditions. Non-invasive, in vivo experiments can be conducted with this experimental apparatus, the potential of which could be usefully exploited in basic biomedical research, prosthesis design, clinical applications, sports medicine and rehabilitation. The device determines the 3D movement of the foot with respect to the shank and evaluates the torques and moments around the three articular axes in relation to any desired angular position of the ankle complex. When integrated with superficial electromyographic techniques and electrical stimulation, it allows the assessment of the functionality of the lower leg in both mechanical and myo-electrical terms. The paper reports the main mechanical and electronic features of the device (high linearity; maximum moment ranges +/- 300 Nm for flexion-extension, +/- 35 Nm for both pronation-supination and internal-external rotation; angular ranges: +/- 100 degrees of dorsi-plantar flexion, +/- 50 degrees of internal-external rotation and prono-supination; linear ranges: +/- 25 mm along each axis). Results from a healthy volunteer, under voluntary or stimulated conditions, helped in testing its operatability, reliability, robustness, repeatability and effectiveness. Preliminary simplified protocols have been also applied to 20 healthy volunteers, and the main results were 80.8 +/- 11.9 degrees of internalexternal rotation, 46.2 +/- 9.1 degrees of prono-supination and 74.6 +/- 13.1 degrees of flexion-extension. Torques and moments were normalised with respect to a body mass index of 30. The maximum plantar flexion moment (57.5 + 21.3 Nm) was measured with the foot at 150 of dorsal flexion; the maximum dorsal flexion moment (50.2 + 20.3 Nm) was measured with the foot at 150 of plantar flexion. PMID- 12892375 TI - Tool-assisted mesh generation based on a tissue-growth model. AB - An heuristic mesh generation technique is proposed that is based on the model of forced particle motion, an edgewise cell-splitting algorithm and a moving tool concept. The method differs from conventional mesh generators in that it uses outward growth of the mesh, in contrast to the inward growth used in traditional meshing techniques. The method does not require prior meshing and patching of two dimensional (2D) boundary surfaces. Instead, it uses a pre-defined skeleton of one-dimensional segments, or an arbitrary tool motion in three-dimensional (3D) space. In this respect, the technique can be considered as a 3D extension of a 2D drawing tool and can find applications in virtual reality systems. The method also guarantees the smoothness of the outer boundary of the mesh at each step of mesh generation, which is not the case with traditional propagating-front methods. The approach is based on the model of tissue growth and is suitable for meshing complex networks of bifurcating branches commonly found in biological structures: blood vessels, lungs, neural networks, plants etc. The generated meshes were used in solving unsteady flow and particle transport problems in lungs. PMID- 12892376 TI - Monitoring the biomechanical response of individual cells under compression: a new compression device. AB - Skeletal muscle cells are sensitive to sustained compression, which can lead to the development of pressure sores. Although it is known that this type of tissue breakdown depends on the magnitude and duration of the applied load, the exact relationship between cell deformation and damage remains unclear. To gain more insight into this process, a method has been developed, that incorporates the use of a new loading device and confocal microscopy. The loading device is able to compress individual cells, either statically or dynamically, while measuring the resulting forces. Experiments can be performed under ideal environmental conditions, comparable with those of a CO2 incubator. First compression experiments on C2C12 mouse myoblasts showed the shape changes that cells undergo during static compression by the loading device. Calculations using the three dimensional confocal images showed no change in volume and an increase in the surface area of the cell as a result of compression. The device presented here provides a useful way to monitor the biomechanical response of skeletal muscle cells during long-term compression experiments. Therefore it will contribute to the knowledge about strain-induced cell damage, as seen in pressure sores and other mechanically induced clinical conditions. PMID- 12892377 TI - Composite lipid polyelectrolyte capsules templated on red blood cells: fabrication and structural characterisation. AB - Charged phospholipids and mixtures of charged phospholipids with zwitterionic lipids were adsorbed onto polyelectrolyte capsules templated on erythrocytes. The assembly was proved by means of electrophoretic mobility measurements, confocal laser scanning microscopy and flow cytometry. Freeze-fracture electron microscopy proved that the phospholipids assemble as bilayers or multilayers. Single particle light scattering showed that bilayers composed of anionic lipids can be intercalated between subsequent polyelectrolyte inter-layers in a regular manner. Neutral lipids can form multilayers. A pronounced decrease in capsule permeability for small polar dyes upon lipid adsorption was followed by confocal laser scanning microscopy. PMID- 12892378 TI - Suppression of neutrophil and endothelial activation by substance P receptor blockade in the Mg-deficient rat. AB - The regulatory role of substance-P (SP) on neutrophil and endothelium activation as well as nitric oxide (NO) production induced by Mg-deficiency was examined. Male Sprague-Dawley rats (180 g) were fed either a Mg-deficient (MgD) or Mg sufficient (MgS) diet for 3 weeks. Enriched neutrophil fractions (> 85%) isolated from whole blood of the Mg-deficient rats displayed an 11-fold (p < 0.001) higher basal superoxide anion producing activity (assayed as SOD-inhibitable cytochrome c reduction) compared to that obtained from the MgS rats. Treatment of the MgD rats with the specific SP-receptor (SPR) blocker, L-703,606 (1 mg/kg/day as s.c. implanted sustained-release pellets) attenuated the superoxide anion producing activity by 75% (p < 0.025). In parallel, circulating prostacyclin (PGI2) level (assayed as 6-keto-PGF-1alpha) was elevated 13-fold in the MgD rats, but was reduced 90% by L-703,606 treatment. Concomitantly, plasma NO products (nitrate + nitrite), which increased 2.2-fold during Mg-deficiency, were completely suppressed by the SPR blockade. When the isolated hearts were subjected to ischemia/ reperfusion stress, NO products were elevated 2.4-fold in the effluent of the MgD group compared to MgS; such heightened NO release was also attenuated after in vivo treatment with the SPR blocker. In conclusion, SP plays a direct role in promoting activation of the neutrophil and endothelium as well as induction of NO production; these processes might participate in the oxidative stress that contributes to the depletion of blood glutathione and cardiac pathology. PMID- 12892379 TI - Protective effect of magnesium and MK 801 on hypoxia-induced hair cell loss in new-born rat cochlea. AB - Hypoxia is a pathogenetic factor in various inner ear diseases, and increasing importance is attached to the protection of the cochlea from traumatic influences. It was recently demonstrated in guinea pigs that magnesium can significantly reduce ischemia- and impulse noise-induced hearing loss. The aim of this study was to evaluate if magnesium has a protective effect on hypoxia induced hair cell loss using an in vitro model of the new-born rat cochlea In view of the NMDA receptor-antagonistic action of magnesium, we tested MK 801, a highly potent and selective non-competitive NMDA receptor antagonist. Organotypic cochlea cultures were exposed to hypoxia (pO2 = 10-20 mm Hg at 37 degrees C) in DMEM medium containing magnesium (0.75 or 3.0 mmol/l) or MK801 (1 or 10 micromol/l) for 24 or 36 h. The cultures were phalloidin-labeled for counting the number of outer and inner hair cells (OHC/IHC). The mean damage in normoxic controls was 1-4%. IHC revealed a significantly higher susceptibility to hypoxia than OHC. In the normal magnesium group (0.75 mmol/l), 36-hour exposure to hypoxia caused a mean loss of about 25% OHC and 60% IHC. In the groups treated with either 3.0 mmol magnesium or 10 microm MK 801, the damage was significantly reduced to about 10% in OHC and 35% in IHC. This study supports previous in vivo observations in the guinea pig demonstrating the protective effects of magnesium on noise-induced impairment of inner ear oxygenation. PMID- 12892380 TI - The influence of alpha-zearalenol on magnesium concentration in cellular subfractions of certain tissues of female rats and their progeny. AB - The purpose of the research work was to study the influence of alpha-zearalenol on magnesium concentration in cellular subfractions of certain tissues of female rats and their progeny. Pregnant female rats received alpha-zearalenol at the doses of: 0.05, 0.5 or 1 mg x kg(-1) b.w. After delivery, brain, liver and kidneys of female rats and their progeny were removed and magnesium concentration was measured in cellular subfractions of these tissues. Alpha-zearalenol reduced Mg-level in all the cellular subfractions of female rat tissues. In progeny tissues, alpha-zearalenol decreased magnesium concentration in nuclear subfraction only. In mitochondrial and cytosol subfractions it increased Mg concentration. The changes in magnesium concentration were depended on alpha zearalenol doses and tissue nature. The magnesium concentration decrease was higher in female rats than in progeny tissues. PMID- 12892381 TI - Enhanced release of IL-1beta and TNF-alpha following endotoxin challenge from rat alveolar macrophages cultured in low-mg(2+) medium. AB - Our previous data have demonstrated that LPS-stimulated alveolar macrophages produce higher levels of IL-1beta and TNF-alpha mRNA in low-Mg(2+) medium than in normal-Mg(2+) medium. In this study, we examined whether the increased mRNA levels are correlated with the release of both cytokines. LPS-stimulated alveolar macrophages released higher amounts of IL-1beta and TNF-alpha in low-Mg(2+) medium than in normal-Mg(2+) medium. The enhanced release of IL-1beta was completely suppressed by pretreatment with verapamil (a calcium entry blocker), U73122 (a phospholipase C inhibitor), W-7 (a calmodulin inhibitor), and curcumin (an activator-protein [AP]-1 inhibitor), and weakly suppressed by dexamethasone (which inhibits nuclear factor [NF]-kappaB and AP-1). On the other hand, the enhanced release of TNF-alpha was completely suppressed by U73122, and strongly suppressed by TMB-8 (which inhibits calcium release from the endoplasmic reticulum) and W-7, and weakly suppressed by pyrrolidine dithiocarbamate (a NF kappaB inhibitor). From these results, we conclude that the enhanced release of IL-1beta and TNF-alpha from LPS-stimulated alveolar macrophages in low-Mg(2+) medium depends partly on the enhanced synthesis of both cytokines, and occurs partly via identical, and partly via different, signaling pathways. PMID- 12892382 TI - Extracellular matrix disturbances in acute myocardial infarction: relation between disease severity and matrix metalloproteinase-1, and effects of magnesium pretreatment on reperfusion injury. AB - PURPOSE: The purpose of this paper is to clarify the relationship between cytokines, matrix metalloproteinase-1 (MMP-1) and severity of acute myocardial infarction (AMI). Additionally, to investigate whether magnesium (Mg) sulfate pretreatment inhibits myocardial damage in coronary reperfusion therapy for patients with AMI. SUBJECTS: At first, 34 patients with AMI were enrolled. Then, the patients were classified into 2 groups with or without congestive heart failure (CHF) (C group and NC group, respectively). Interleukin 6 (IL-6), MMP-1 and the hemodynamic parameters were measured. Second, 36 AMI patients treated with coronary reperfusion therapy were enrolled. Patients were divided into 2 groups (18 patients each) as the non-pretreated group (Control group) and the group pretreated with intravenous Mg sulfate (0.27 mmol/kg) (Mg group). IL-6, MMP 1 and the indexes of reperfusion injury were evaluated. RESULTS: There were positive correlations between peak MMP-1 level, and peak creatine kinase value and pulmonary capillary wedge pressure and peak IL-6 level (r = 0.43, r = 0.70, and r = 0.60, respectively) in all patients. There were negative correlations between peak MMP-1 level and left ventricular ejection fraction and cardiac index (r = - 0.52 and r = - 0.55, respectively). The peak blood IL-6 and MMP-1 level increased in AMI, particularly in patients with CHF (C group vs NC group; 130 vs 51 pg/mL and 37 vs 18 ng/mL, both p < 0.01). Additionally, peak IL-6 and peak MMP 1 in the Mg group were lower than those of the control group (39 vs 92 pg/mL and 16 vs 20 ng/mL, p < 0.05 and p = 0.09, respectively). The incidence of reperfusion injury including reperfusion arrhythmia and transient exacerbation of ST elevation in the Mg group was lower than that of control group (17 vs 78% and 2.5 vs 4.7 mm, p < 0.01 and p = 0.08, respectively). CONCLUSION: These results may suggest that the severity of AMI is reflected by the blood IL-6 and MMP-1 levels and that pretreatment with Mg administration protects the myocardium of patients with AMI from reperfusion injury induced by IL-6 and MMP-1. PMID- 12892383 TI - Lower whole blood ionized magnesium concentrations in hypocalcemic infants of gestational diabetic mothers. AB - Infants of insulin dependent (class B and above) diabetic mothers (IDM's) have a high rate of neonatal hypocalcemia (NHC) and hypomagnesemia. We carried out this study to test the hypotheses that: (1) infants of gestational diabetic (class A) mothers (IGDM's) are also at risk for NHC and (2) NHC in IGDM's relates to decreased whole blood Mg(2+) concentration. Thirty one term infants born to gestational diabetic mothers of classes A1 (diet controlled, n = 23) and A2 (requiring insulin, n = 8) of White's classification, were compared at 24 +/- 2 hours of age to 32 healthy, appropriate for gestational age controls, born after uncomplicated pregnancy, labor and delivery. Whole blood Mg(2+) and Ca(2+) were measured using an ion-specific electrode (Nova 8, Nova biomedical, Waltham, MA). The rate of NHC was higher in the IGDM group as compared to the control group (9 out of 31 [29%] vs. 1 out of 32 [3.1%] infants; p < 0.01). Whole blood Mg(2+) was lower in the IGDM group than in controls (p < 0.05). In multiple regression analysis, when Ca(2+) was used as the dependent variable and Mg(2+), diabetes class, gestational age, macrosomia and one minute Apgar scores were the independent variables, only Mg(2+) and diabetes class were significant (R2 = 0.4374; p < 0.01). Our results are consistent with the theory that Mg deficiency plays a role in NHC encountered in IGDM's, similar to what occurs in infants of insulin-dependent diabetic mothers. PMID- 12892384 TI - Magnesium-orotate supplementation for idiopathic infertile male patients: a randomized, placebo-controlled clinical pilot study. AB - A randomized, placebo-controlled clinical pilot study was performed in order to examine the effect of magnesium-orotate in male idiopathic infertility. Ten males were treated daily for 90 consecutive days with 3000 mg magnesium-orotate (Magnerot) tablets (Group M). As a control, ten other males were treated in the same way with placebo (Group P). Conventional microscopic sperm characteristics (sperm concentration, motility ratio, total number of motile sperm cells, normal morphology ratio), plus total and ionized magnesium levels in seminal plasma and blood serum were evaluated both prior to treatment and on day 90, at the conclusion of the study. No significant changes in sperm characteristics, blood ionized or total Mg, or ejaculate total Mg levels were detected. However, ejaculate ionized Mg levels increased in Group M from 0.18 +/- 0.05 to 0.30 +/- 0.05 (mmol/l; mean +/- SD, p < 0.05). Within the observation period of 3 months, one pregnancy occurred in the partner of a male from Group M. In conclusion, magnesium-orotate treatment at a dose of 3000 mg/day leads neither to a significant improvement of sperm variables nor does it increase the pregnancy rates of female partners of treated males as compared to those of controls. Thus, magnesium-orotate treatment was not shown to be effective therapy for idiopathic male infertility. PMID- 12892385 TI - Antiphospholipid syndrome: a consensus for treatment? PMID- 12892386 TI - Systemic antiphospholipid syndrome. AB - The antiphospholipid syndrome (APS) was reported in the early 1980s as a triad of manifestations. Since than it became one of the most systemic conditions. Almost any organ and tissue may be involved in the disease, including the brain, the heart, the placenta, the endocrine system, the blood, the kidneys and many more. In the article it is suggested to add to the syndrome the word systemic in analogy to its sister condition, systemic lupus erythematosus. PMID- 12892387 TI - Prophylaxis of the antiphospholipid syndrome: a consensus report. AB - Hypothetical circumstances that may require prophylaxis for a potential antiphospholipid syndrome (primary prophylaxis), or in some instances when there already had been some manifestations ofthe syndrome (secondary prophylaxis), were presented to a panel of experts for their consideration on potential prophylactic intervention. These were subsequently presented to the participants in the First International Consensus on Treatment of the Antiphospholipid Syndrome. In most instances there was consensus in adding low dose aspirin, an exception being aspirin allergy when other antiaggregants could be used in nonpregnant subjects. General measures to prevent thrombosis and other vasoprotective actions should also be provided. Higher risk of fetal loss or thrombosis called for anticoagulation with coumadin in nonpregnant subjects or subcutaneous low molecular weight heparin in pregnant ones. When indicated, prophylaxis of the antiphospholipid syndrome should be provided in systemic lupus erythematosus patients who are being treated for their disease. In no instance should corticosteroids or immunosuppresants be given as prophylactic of an antiphospholipid syndrome. PMID- 12892388 TI - Venous thromboembolism in the antiphospholipid syndrome: management guidelines for secondary prophylaxis. AB - Venous thromboembolism (VTE) in patients suffering from the antiphospholipid syndrome (APS) has been reported in almost any location of the vessel tree and the risk of recurrences has been found in several studies to be more closely associated with the presence of lupus anticoagulant than with the positivity for anti-cardiolipin antibodies. The thrombophilic state of APS raises the problem of the secondary prophylaxis to avoid VTE recurrences. For APS patients with VTE, published data appear to support a longer warfarin treatment if compared with the standard management of antiphospholipid (aPL)-negative patients with VTE. The question of how long oral anticoagulant treatment should be continued for APS patients, however, remains unanswered. Concerning the intensity of anticoagulation, several authors recommend a target international normalized ratio (INR) between 3.0 and 4.0 to efficiently protect from VTE recurrences. A recent decision analysis study does support such a suggestion. On the contrary, in a few prospective studies regimens with lower target INRs appear to be effective, and some authors therefore recommend a target INR of between 2.0 and 3.0. Specific large and prospective trials are needed to address this question. Until such information becomes available, individualized treatment according to the patient's individual risk factors for both bleeding and thrombosis is the general practice. PMID- 12892389 TI - Stroke and the antiphospholipid syndrome: consensus meeting Taormina 2002. AB - Ischaemic stroke is the only neurological manifestation accepted as a clinical diagnostic criterion for the antiphospholipid syndrome (APS). This association is reasonably well established in patients first diagnosed with APS but is less clear in randomly selected stroke patients who test positive on one occasion for antiphospholipid antibodies and who have no other evidence of systemic autoimmune disease. We propose a grading system that posits stroke to be definitely, likely or possibly associated with antiphospholipid antibodies (aPL). Further, there are limited prospective data to determine appropriate treatment. There is controversy as to whether the presence of aPL even increases risk of a recurrent stroke or other thromboembolic event, although data point to persistent medium-high titre aCL and/or LA as risk factors for recurrence. In the absence of data to guide clinicians on the best treatment, we cannot make strong recommnendations as to optimal therapy, nor can we propose clear consensus treatment guidelines. PMID- 12892390 TI - Non-stroke neurological syndromes associated with antiphospholipid antibodies: evaluation of clinical and experimental studies. AB - Although many types of neurological disorders and events have been described in association with antiphospholipid antibodies (aPL) and the antiphospholipid syndrome (APS), only ischaemic stroke is reasonably well established and accepted as a diagnostic criterion for the syndrome. We propose to evaluate, classify and rank the association of other neurological manifestations as possible, probable, or definite according to the data available from clinical studies and animal models. By these criteria, none of the neurological disorders or events such as epilepsy, psychiatric disease, dementia, transverse myelitis, multiple sclerosis like disease, chorea, migraine, Guillian-Barre syndrome, and sensory-neural hearing loss, can be definitely associated with aPL or APS. PMID- 12892391 TI - Cardiac disease in the antiphospholipid syndrome: recommendations for treatment. Committee consensus report. AB - The Committee reviewed cardiac involvement in the antiphospholipid antibody syndrome. The Committee's recommendations are: Valve abnormalities: anticoagulation is recommended for symptomatic patients with valvulopathy. Prophylactic antiplatelet therapy may be appropriate for asymptomatic patients (recommended by 13/17 experts in an independent review). Committee members disagreed whether corticosteroid therapy is helpful, but agree that distinguishing among presumptive valvulitis (valve thickening on echocardiogram), valve deformity and vegetations is important, as treatment implications may differ. Occlusive arterial disease (angina, myocardial infarction): the Committee recommends aggressive treatment of all risk factors for atherosclerosis (hypertension, hypercholesterolaemia, smoking) and liberal use of folic acid, B vitamins and cholesterol-lowering drugs (preferably statins). Hydroxychloroquine for cardiac protection in APS patients may be considered. The Committee also recommends warfarin anticoagulation for those who have suffered thrombosis in the absence of atherosclerosis, but recognizes that developing data may support the use of antiplatelet agents instead. Intracardiac thrombi: the Committee recommends intensive warfarin anticoagulation, and consultation with cardiac surgeons when appropriate. Ventricular dysfunction: the Committee has no recommendations on this aspect of cardiac disease. Pulmonary hypertension: the Committee recommends intensive anticoagulation with warfarin and clinical trials of bosentan, epoprostenol and other new agents. PMID- 12892392 TI - Treatment of pregnant patients with antiphospholipid syndrome. AB - Antiphospholipid Syndrome (APS) has been widely recognized as a risk factor for the recurrence of both thrombosis and pregnancy losses; however the optimal treatment of patients is debatable. The aim of this paper was to establish a consensus among experts on the treatment of APS in pregnancy. A questionnaire that described possible different clinical situations was sent to the International Advisory Board of the 10th International Congress on Antiphospholipid Antibodies. Sixteen experts from different medical branches and different geographic areas sent their replies. The consensus was that treatment for APS pregnant patients should be low molecular weight heparin (LMWH) and low dose aspirin (LDA). The dosage, and frequency of LMWH depends on different situations, including the body weight and past history. Patients with previous thromboses usually receive two injections per day. Warfarin can also be used from 14 to 34 weeks, for patients with previous stroke or severe arterial thromboses. The use of intravenous immunoglobulin (IVIG) seems to be restricted to patients with pregnancy losses despite conventional treatment. The experts usually advised barrier methods of contraception, intrauterine device (if the patient is not taking corticosteroids) or progestins. Oral contraception with oestrogens was usually avoided. PMID- 12892393 TI - Catastrophic antiphospholipid syndrome: international consensus statement on classification criteria and treatment guidelines. AB - The term 'catastrophic' antiphospholipid syndrome (APS) is used to define an accelerated form of APS resulting in multiorgan failure. Although catastrophic APS patients represent less than 1% of all patients with APS, they are usually in a life-threatening medical situation that requires high clinical awareness. The careful and open discussion of several proposals by all participants in the presymposium workshop on APS consensus, held in Taormina on occasion of the 10th International Congress on aPL and chaired by Munther A Khamashta and Yehuda Shoenfeld (29 September 2002), has allowed the acceptation of a preliminary set of classification criteria. On the other hand, the optimal management of catastrophic APS must have three clear aims: to treat any precipitating factors (prompt use of antibiotics if infection is suspected, amputation for any necrotic organ, high awareness in patients with APS who undergo an operation or an invasive procedure), to prevent and to treat the ongoing thrombotic events and to suppress the excessive cytokine 'storm'. Anticoagulation (usually intravenous heparin followed by oral anticoagulants), corticosteroids, plasma exchange, intravenous gammaglobulins and, if associated with lupus flare, cyclophosphamide, are the most commonly used treatments for catastrophic APS patients. PMID- 12892394 TI - Activation of complement mediates antiphospholipid antibody-induced pregnancy loss. AB - Although it is clear that the specific antigenic reactivity of antiphospholipid (aPL) antibodies is critical to their effect, the pathogenic mechanisms that result in injury in vivo are incompletely understood. We hyphothesized that aPL antibodies targeted to the placenta activate complement locally, generating split products that mediate placental injury and lead to foetal loss and growth retardation. To test this hypothesis, we used a murine model of APS in which pregnant mice are injected with human IgG containing aPL antibodies. Mice treated with inhibitors of complement activation and mice deficient in complement components were protected from aPL antibody-induced foetal damage. Although the cause of tissue injury in this disease is probably multifactoral, we have shown that complement activation is an absolute requirement for foetal loss and growth restriction and, therefore, thatthis pathway acts upstream of other important effector mechanisms. Identification of complement activation as a mechanism that is necessary for aPL-induced tissue damage and definition ofthe complement components necessary to trigger such injury is likely to lead to a better understanding of the pathogenesis of vascular and tissue injury in SLE and to new and improved treatments. PMID- 12892395 TI - Probing antiphospholipid-mediated thrombosis: the interplay between anticardiolipin antibodies and endothelial cells. AB - The association of antiphospholipid (aPL) antibodies with thrombosis in patients with antiphospholipid syndrome (APS) is well documented in humans and in animal studies. However, the mechanisms by which aPL antibodies induce thrombosis are the subject of much current study. It has been suggested that aPL may activate endothelial cells (ECs), thus creating a hypercoagulable state that precedes and contributes to thrombosis in patients with APS. Several studies have shown that aPL upregulate ECs' adhesion molecules (CAMs): intercellular cell adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1) and E-selectin (E sel) or induce tissue factor (TF) in monocytes in vitro. Similarly, the incubation of EC with antibodies reacting with beta2glycoprotein I (beta2GPI) has been shown to induce EC activation with concomitant upregulation of CAMs, IL-6 production and alteration of prostaglandin metabolism. Our group has shown that aPL-mediated upregulation of adhesion molecules on ECs correlates with an increased adhesion of leukocytes to endothelium in the microcirculation of mouse cremaster muscle, a n indication of EC activation in vivo, andwith enhanced thrombosis in vivo. In another series of studies, investigators have shown that upregulation of expression of adhesion molecules by some murine monoclonal anti beta2glycoprotein I (anti-beta2GPI) antibodies correlated with fetal resorption in mice in vivo. More recently, one study showed that the anti hypercholesterolaemic drug fluvastatin inhibited the aPL-mediated enhanced adhesion of monocytes to ECs in vitro. Data from our laboratories indicate that fluvastatin also reverses thrombus formation and activation of EC induced by aPL in an in vivo mouse model. As additional support for the hypothesis that aPL antibodies activate ECs and may create an hypercoagulable state in APS patients, two recent studies indicated that levels of soluble ICAM-1 and VCAM-1 were significantly increased in the plasma of patients with APS and recurrent thrombosis. Furthermore, studies utilizing knockout mice and specific monoclonal anti-VCAM-1 antibodies have demonstrated that expression of ICAM-1, P-selectin, E selectin and VCAM-1 are important in in vivo aPL-mediated thrombosis and EC activation in mice. Recent data suggests that aPL antibodies also induce expression of TF not only in monocytes but in ECs. Hence, the interference of aPL with the TF mechanism may be another important mechanism by which these antibodies create a hypercoagulable state and prone patients to thrombosis. Specifically, how aPL alters EC activation state and the molecular and intracellular mechanisms involved have not yet been defined. APL may interact with specific cell surface receptors (proteins and/or lipids) induce signals that have consequences downstream, and that ultimately will result in upregulation of cell surface proteins (i.e., CAMs and TF) and subsequently induce EC activation. In that regard, our group recently showed that aPL-mediated upregulation of adhesion molecules in ECs is preceded by activation of the nuclear factor kappa B (NFkappaB). Other intracellular mechanisms triggered by aPL are not completely understood and are the subject of current investigation. In conclusion, studies suggest that activation of ECs by aPL is an important mechanism that may precede thrombus formation in patients with APS. Hence, the interplay between aPL antibodies and ECs is important inthe pathogenesis of thrombosis in APS. PMID- 12892396 TI - TNFalpha DNA vaccination prevents clinical manifestations of experimental antiphospholipid syndrome. AB - Naked DNA encoding TNFalpha was introduced to BALB/c mice with experimental antiphospholipid syndrome (APS) induced by beta2GPI. Administration of naked DNA encoding TNFalpha resulted in the generation of immunological memory to its gene product, associated with elevated circulating anti-TNFalpha antibodies. Enriched IgG fraction of the mouse anti-TNFalpha was biologically active since it prevented endothelial cell activation by TNFalpha e.g., inhibition of monocyte adhesion to activated endothelial cells (HUVEC). Mice immunized with beta2GPI, vaccinated with TNFalpha DNA at an early stage of disease development, showed decreased titres of circulating anti-beta2GPI antibodies as compared to the group of mice vaccinated with a control naked DNA. The reduction of antiphospholipid antibody production was followed by amelioration of the foetal loss, increased platelet count to normal values as well as normalization of the prolonged aPTT. APS mice which were introduced to the TNFalpha DNA vector at a later stage of the disease development, showed less improvement in their clinical manifestations. The current study suggests a way in which a DNA vaccine can be employed for induction of a protective immunity in experimental APS. PMID- 12892397 TI - Oxidized low-density lipoprotein as a risk factor of thrombosis in antiphospholipid syndrome. AB - Beta2-Glycoprotein I (beta2-GPI) is a major antigen for anticardiolipin antibodies (aCL, Abs) present in patients with antiphospholipid syndrome (APS). We recently reported that oxidized LDL(oxLDL) is subsequently targeted by beta2 GPI and anti-beta2-GPI auto-Abs and that-carboxyl variants of 7-ketocholesteryl esters, such as 7-ketocholesteryl-9-carboxynonanoate (oxLig-1) and 7 ketocholesteryl-12-carboxy (keto) octadodecanoate (oxLig-2), are ligands for beta2-GPI (J Lipid Re 2001; 42: 697; J Lipid Res 2002; 43: 1486). These beta2-GPI ligands provide an electrostatic interaction between oxLDL and beta2-GPI followed by forming stable complexes (such as Schiff base adducts). The omega-carboxyl function in these ligand is responsible for beta2-GPI binding to oxLDL and the oxLDL-beta2-GPI complexes are anti-beta2-GPI auto-Ab-dependently taken up by macrophages (i.e., by phagocytosis). Our recent observations are consistent with the evidence that beta2-GPI co-localizes with lymphocytes and mononuclear cells in human athero-plaques. Thus, autoimmune thrombogenesis (atherogenesis) is linked to interaction of anti-beta2-GPI Abs with the beta2-GPI-oxLDL complexes. We propose an alternative idea, that an immune response against the beta2-GPI oxLDL complexes may be involved in mechanisms in the development of atherosclerosis, which has been explained by the theory of 'the response to injury'. PMID- 12892398 TI - Antiphospholipid antibodies and renal transplantation: a risk assessment. AB - Data from 110 transplanted patients show that the presence of antiphospholipid antibodies (aPL) at the time of transplantation is an important risk factor for early renal allograft failure. Sera were tested for IgG, IgM and IgA to CL, PS, PE and PC. Haemodialysis patients had a significantly higher incidence of aPL compared to patients who did not receive haemodialysis (P = 0.0171). aPL-positive ESRD patients on peritoneal dialysis (CAPD) or who had never received haemodialysis were at maximal risk; 100% failure (P = 0.0022). aPL-positive patients receiving haemodialysis were not at such risk. Biopsy findings from the failed kidneys show abundant fibrin deposition in the microvasculature. Serial blood samples from transplanted patients showed aPL titres to decrease immediately after transplant and increase after removal of the failed graft, indicating that aPL specifically target the allografts. To confirm this, we were able to isolate aPL from a failed graft after transplant nephrectomy. Ninety seven per cent of the aPL-positive patients' historic pre-transplant serum samples demonstrated the presence of the same aPL specificity detected in the final crossmatch sera. The exposure to heparin during haemodialysis suggested to us that heparin reduces the risk of clotting in aPL positive transplant candidates. To lessen the risk of graft loss in aPL positive kidney transplant patients (including CAPD), subcutaneous heparin was administered peri- and post operatively. To date, none of the heparin-treated aPL-positive transplanted patients suffered an early graft loss. Further, they experienced fewer rejection episodes requiring biopsy and thus are prescribed less steroid therapy than patients not treated with heparin. PMID- 12892399 TI - HLA-DPB1 alleles association of anticardiolipin and anti-beta2GPI antibodies in a large series of European patients with systemic lupus erythematosus. AB - Our objective was to determine the HLA-DPB1 allele associations of anticardiolipin (aCL) and anti-beta2GPI (a(beta)2GPI) antibodies, and of clinical manifestations of the antiphospholipid syndrome (APS), in systemic lupus erythematosus (SLE). We studied 577 European patients with SLE. aCL and a(beta)2GPI antibodies were measured by ELISA. Molecular typing of HLA-DPB1 locus was performed by polymerase chain reaction-sequence specific oligonucleotide probe (PCR-SSOP) method. aCL showed positive association with -DPB1*1501 (P = 0.005, OR = 7.4), and -DPB1*2301 (P = 0.009, OR = 3.3). a(beta)2GPI showed positive association with -DPB1*0301 (P = 0.01, OR = 1.9), and -DPB1*1901 (P = 0.004, OR = 8.1). In addition, livedo reticularis was associated with -DPB1*1401, and Raynaud's phenomenon with -DPB1*2001. In conclusion, HLA-DPB1 locus may contribute to the genetic predisposition to develop antiphospholipid antibodies and clinical manifestations of the APS in patients with SLE. PMID- 12892400 TI - Primary anetoderma: a cutaneous sign of antiphospholipid antibodies. AB - Although a few reports in recent years have suggested that patients with antiphospholipid antibodies (aPL) are prone to developing primary anetoderma (PA), it is still unclear how often aPL are detected in unselected PA patients. We studied nine consecutive PA patients for the presence of autoimmune antibodies and disorders in general and the presence of aPL in particular. Six of the nine patients had clinical evidence of associated autoimmune disorders (Graves'disease and autoimmune haemolysis in one, systemic scleroderma in one, Hashimoto's thyroiditis in one, alopecia areata in one) and/or signs of hypercoagulability (recurrent fetal loss in two, recurrent stokes in one, recurrent deep vein thrombosis in one). In four ofthese six patients the onset of PA preceded these signs. Positive aPL was found in all: anticardiolipin (aCL) in six, anti-beta2 glycoprotein-I (a(beta)2GPI) in six and lupus anticoagulant (LAC) in four. The most frequent isotype was IgA. Among other autoantibodies found the most frequently was antinuclear antibodies. Four ofthe nine patients fulfilled the criteria for antiphospholipid syndrome (APS). It is concluded that PA is an important cutaneous sign for autoimmune disorders in general and the presence of aPL in particular. Hence, the work-up of these patients should include testing for LAC as well as for all different isotypes ofaCL and a(beta)2GPI. We recommend that PA be added to the list of the cutaneous manifestations of APS. PMID- 12892401 TI - Adrenal involvement in the antiphospholipid syndrome. AB - Although Addison disease and hypoadrenalism are rare in patients with systemic lupus erythematosus (SLE), early reports of the association suggested the presence of antiphospholipid antibodies (aPL) in these patients. Data from literature reveal that adrenal failure was present in between 10 and 26% of patients with catastrophic APS and that one-third of patients presented with adrenal involvement during the course of catastrophic APS. Adrenal involvement may be the first clinical manifestation of this syndrome, whereas a few patients may have a history of Addison's disease in the past. The pathological mechanisms involved in the production of adrenal insufficiency in APS are still not clearly understood, but the hypercoagulable state in these patients supports the concept that adrenal haemorrhagic infarction may possibly be related to adrenal vein thrombosis. In the present article,we review the pathogenic mechanisms and main clinical, laboratory and treatment features of patients suffering adrenal involvement with aPL to support the idea that APS leads to the development of adrenal insufficiency. PMID- 12892402 TI - The effects of antiphospholipid antibodies obtained from women with SLE/APS and associated pregnancy loss on rat embryos and placental explants in culture. AB - Recurrent fetal loss occurs in approximately 1% of women. Autoimmune causes have been suggested as a factor in some of these cases. High rates of intrauterine fetal growth retardation and increased incidence of prematurity is associated with systemic lupus erythematosus (SLE) and the antiphospholipid syndrome (APS). We found in previous studies that sera from SLE/APS patients when used as a culture medium for rat embryos were found to reduce embryonic growth and development, induce a high rate of embryonic anomalies and death and damage the yolk sac morphologically and functionally. In order to investigate the direct effect of IgG purified from women with SLE/APS on the growth and viability of embryos, we cultured 11.5-day-old rat embryos in their yolk sacs in the presence of IgG purified from SLE/APS patients with recurrent pregnancy loss (RPL). The IgG affected directly the embryo and yolk sac, reducing their growth. The purified IgG positive for anticardiolipin/anti-DNA antibodies reduced yolk sac and embryonic growth more than sera negative for these antibodies but positive for antiphosphatydilserine and for antilaminin. Monoclonal antiphosphatydilserine reduced yolk sac growth but the embryos remained intact. Following the observed damage to the yolk sac we cultured human placental explants at 5.5-8 weeks of pregnancy in sera from SLE/APS patients for 96 hours and found that these sera reduced placental trophoblastic cell growth, reduced their proliferation rate and increased their rate of apoptosis. Successful treatment of the women resulted in a correction of the damage induced in the cultured rat embryos and in the cultured placental explants. PMID- 12892404 TI - Histochemical alterations of re-innervated rat extensor digitorum longus muscle after end-to-end or graft repair: a comparative histomorphological study. AB - Changes in the histochemical profile of 43 rat extensor digitorum longus muscles undergoing de-innervation and re-innervation were recorded. Assessment of fibre type composition and muscle fibre cross-sectional area was performed at 15, 30, 90 and 180 days post operative (p.o.) after either primary end-to-end repair or autologous graft repair of the common peroneal nerve (n = 5 per time point and type of repair). The size and histochemical profile of single muscle fibres were analysed by computer-assisted quantification on the basis of their myofibrillar ATPase (pH 4.3) and succinate dehydrogenase (SDH) activities in serial, whole muscle cross-sections. Accordingly, four muscle-fibre types could be functionally identified: (1) slow oxidative (SO, type I); (2) fast-oxidative glycolytic (FOG, type IIA); (3) fast glycolytic (FG, type IIB); and (4) succinate dehydrogenase intermediate (SDH-INT). At 15 days following end-to-end repair, the SDH-INT muscle fibre type was observed. By contrast, 15 days following graft repair, no changes in fibre type composition were observed (vs. control). At 30 days p.o. in the group that received end-to-end repair, type SDH-INT reached its maximum and was significantly higher than in the group that underwent graft repair. At 90 days p.o., the amount of SDH-INT fibres declined after end-to-end repair, but it was still significantly higher than in the group treated with a nerve graft. The increase of the SDH-INT fibre type was mirrored by a proportional disappearance of FG and FOG fibres. These changes were time-dependent, not reversible at 180 days p.o and largely blunted after nerve graft. Muscle-fibre size decreased at 15 and 30 days after both types of nerve repair. This decrease was transient and reversible within 90 days p.o. These findings reflect the fact that the reorganization of the histochemical profile in re-innervated muscles is both time dependent and long lasting. The degree of this reorganization is significantly higher after end-to-end repair than after graft repair. PMID- 12892405 TI - Ultrastructural distribution of glycinergic and GABAergic neurons and axon terminals in the rat dorsal cochlear nucleus, with emphasis on granule cell areas. AB - A knowledge of neurotransmitters in the neurons of the rat cochlear nuclear complex is of importance in understanding the function of auditory circuits. Using post-embedding ultrastructural immunogold labelling, the distribution of glycinergic and GABAergic neurons and axonal terminals has been studied in the molecular, fusiform and polymorphic layers of the rat dorsal cochlear nucleus (DCN). This technique is not limited by the penetration of antibodies into the nervous tissue as in pre-embedding methods, and allows a fine neurochemical mapping of the nervous tissue. Numerous glycinergic and GABAergic axon terminals contain pleomorphic and flat synaptic vesicles, and are present in all layers (1, 2, 3) of the dorsal cochlear nucleus. Glycine and GABA-negative large terminals (mossy fibres) are mainly seen in granule cell areas of layer 2 (fusiform layer). Mossy fibres contact the dendrites of GABA- and glycine-negative granule cells and of the few unipolar brush cells (excitatory neurons). The least common cells in the granule cell areas are GABAergic and glycinergic Golgi-stellate neurons. In unipolar brush cells, aggregations of vesicles seem to be the origin of their characteristic ringlet-bodies. Golgi-stellate cells send their inhibitory terminals to the dendrites of granule and unipolar brush cells, occasionally directly to mossy fibres. Small or (less frequently) large GABAergic terminals contact the soma or the main dendrite of unipolar brush cells. The circuit of a hypothetical functional unit of neurons in the DCN is proposed. The inputs from auditory tonotopic or non-auditory non-tonotopic mossy fibres eventually reach pyramidal cells through axons from the granule cells or unipolar brush cells. Pyramidal cells convey an excitatory signal from the DCN to higher mesencephalic nuclei for further elaboration of the acoustic signal. PMID- 12892406 TI - Magnetic resonance images of the brain of a dwarf sperm whale (Kogia simus). AB - Cetacean (dolphin, whale and porpoise) brains are among the least studied mammalian brains because of the difficulty of collecting and histologically preparing such relatively rare and large specimens. Among cetaceans, there exist relatively few studies of the brain of the dwarf sperm whale (Kogia simus). Magnetic resonance imaging (MRI) offers a means of observing the internal structure of the brain when traditional histological procedures are not practical. Therefore, MRI has become a critical tool in the study of the brain of cetaceans and other large species. This paper represents the first MRI-based anatomically labelled three-dimensional description of the dwarf sperm whale brain. Coronal plane sections of the brain of a sub-adult dwarf sperm whale were originally acquired and used to produce virtual digital scans in the other two orthogonal spatial planes. A sequential set of images in all three planes has been anatomically labelled and displays the proportions and positions of major neuroanatomical features. PMID- 12892407 TI - Roles of HNK-1 carbohydrate epitope and its synthetic glucuronyltransferase genes on migration of rat neural crest cells. AB - HNK-1 carbohydrate epitope is localized on the surface of avian neural crest cells (NCCs), and is necessary for their migration. However, it is still disputed whether the epitope works in similar ways in mammalian embryos. In this study, we found that HNK-1 carbohydrate epitope was specifically detected in some of the cranial ganglia, migrating trunk NCCs and some non-NCC derivatives in the rat embryo. Two genes encoding glucuronyltransferases that synthesize the HNK-1 epitope in vitro (GlcAT-P and GlcAT-D) were recently identified in the rat. Interestingly, the NCCs in the cranial ganglia expressed the GlcAT-D gene, whereas the migrating trunk NCCs expressed the GlcAT-P gene. To investigate in vivo functions of the GlcATs in the NCC migration further, we overexpressed GlcAT genes by electroporation in the cranial NCCs in cultured rat embryos. Transfection of both GlcAT genes resulted in efficient synthesis of the HNK-1 epitope in the NCCs. GlcAT-P overexpression increased distance of cranial NCC migration, whereas GlcAT-D overexpression did not show this effect. Our data suggest that the HNK-1 epitope synthesized by different GlcATs is involved in migration in the sublineages of the NCCs in the rat embryo, and that GlcAT-P and GlcAT-D mediate different effects on the NCC migration. PMID- 12892403 TI - Neurotrophins and the immune system. AB - The neurotrophins are a family of polypeptide growth factors that are essential for the development and maintenance of the vertebrate nervous system. In recent years, data have emerged indicating that neurotrophins could have a broader role than their name might suggest. In particular, the putative role of NGF and its receptor TrkA in immune system homeostasis has become a much studied topic, whereas information on the other neurotrophins is scarce in this regard. This paper reviews what is known about the expression and possible functions of neurotrophins and their receptors in different immune tissues and cells, as well as recent data obtained from studies of transgenic mice in our laboratory. Results from studies to date support the idea that neurotrophins may regulate some immune functions. They also play an important role in the development of the thymus and in the survival of thymocytes. PMID- 12892409 TI - Force- and moment-generating capacities of muscles in the distal forelimb of the horse. AB - A detailed musculoskeletal model of the distal equine forelimb was developed to study the influence of musculoskeletal geometry (i.e. muscle paths) and muscle physiology (i.e. force-length properties) on the force- and moment-generating capacities of muscles crossing the carpal and metacarpophalangeal joints. The distal forelimb skeleton was represented as a five degree-of-freedom kinematic linkage comprised of eight bones (humerus, radius and ulna combined, proximal carpus, distal carpus, metacarpus, proximal phalanx, intermediate phalanx and distal phalanx) and seven joints (elbow, radiocarpal, intercarpal, carpometacarpal, metacarpophalangeal (MCP), proximal interphalangeal (pastern) and distal interphalangeal (coffin)). Bone surfaces were reconstructed from computed tomography scans obtained from the left forelimb of a Thoroughbred horse. The model was actuated by nine muscle-tendon units. Each unit was represented as a three-element Hill-type muscle in series with an elastic tendon. Architectural parameters specifying the force-producing properties of each muscle tendon unit were found by dissecting seven forelimbs from five Thoroughbred horses. Maximum isometric moments were calculated for a wide range of joint angles by fully activating the extensor and flexor muscles crossing the carpus and MCP joint. Peak isometric moments generated by the flexor muscles were an order of magnitude greater than those generated by the extensor muscles at both the carpus and the MCP joint. For each flexor muscle in the model, the shape of the maximum isometric joint moment-angle curve was dominated by the variation in muscle force. By contrast, the moment-angle curves for the muscles that extend the MCP joint were determined mainly by the variation in muscle moment arms. The suspensory and check ligaments contributed more than half of the total support moment developed about the MCP joint in the model. When combined with appropriate in vivo measurements of joint kinematics and ground-reaction forces, the model may be used to determine muscle-tendon and joint-reaction forces generated during gait. PMID- 12892408 TI - Muscle satellite (stem) cell activation during local tissue injury and repair. AB - In post-mitotic tissues, damaged cells are not replaced by new cells and hence effective local tissue repair mechanisms are required. In skeletal muscle, which is a syncytium, additional nuclei are obtained from muscle satellite (stem) cells that multiply and then fuse with the damaged fibres. Although insulin-like growth factor-I (IGF-l) had been previously implicated, it is now clear that muscle expresses at least two splice variants of the IGF-I gene: a mechanosensitive, autocrine, growth factor (MGF) and one that is similar to the liver type (IGF IEa). To investigate this activation mechanism, local damage was induced by stretch combined with electrical stimulation or injection of bupivacaine in the rat anterior tibialis muscle and the time course of regeneration followed morphologically. Satellite cell activation was studied by the distribution and levels of expression of M-cadherin (M-cad) and related to the expression of the two forms of IGF-I. It was found that the following local damage MGF expression preceded that of M-cad whereas IGF-IEa peaked later than M-cad. The evidence suggests therefore that an initial pulse of MGF expression following damage is what activates the satellite cells and that this is followed by the later expression of IGF-IEa to maintain protein synthesis to complete the repair. PMID- 12892410 TI - The peri-albumen layer: a novel structure in the envelopes of an avian egg. AB - The present paper describes a novel structure, termed the peri-albumen layer, in the egg-envelopes of the quail Coturnix japonica. It reacts with Alcian blue and exists between the egg white and the shell membrane. Ultrastructurally, it is of fine granular structure and forms a fenestrate sheet, the width of which is 190 nm or less. Isolated materials of the peri-albumen layer include an Alcian-blue positive polysaccharide of 260 kDa, and three glycoproteins of 160, 108 and 52 kDa. The layer is supplied to an egg when it passes through the magnum-isthmus junction, the normalized length of which is 0.62-0.63 of the oviduct. The mucosa of the junction consists exclusively of a luminal epithelium. It is apparently distinct from the mucosa of the magnum and the isthmus, which consist of a luminal epithelium and tubular glands. The luminal epithelium of the magnum isthmus junction stains prominently with Alcian blue and consists of alternately distributed ciliated cells and granular cells. Immunohistochemistry with an antiserum raised against the materials of the peri-albumen layer revealed the staining of the peri-albumen layer of the egg, and secretory cells of the luminal epithelium at the magnum-isthmus junction. It was concluded that the materials of the peri-albumen layer are produced by secretory cells at the magnum-isthmus junction of the oviduct. PMID- 12892411 TI - The detection of mature T- and B-cells during development of the lymphoid tissues of the tammar wallaby (Macropus eugenii). AB - The distribution of T- and B-cells in the developing lymphoid and immunohaematopoietic tissues of the tammar wallaby were investigated using antibodies to the mature cell surface markers, CD3, CD5 and CD79b. In the thymus, CD3- and CD5-positive T-cells were first observed at day 12 postpartum whilst rare B-cells were first detected at day 23. Both T- and B-lymphocytes were first stained on day 21 postpartum in the spleen and day 24 in lymph nodes. In one sample from a 7-day-old animal, rare CD79b-positive (CD79b+) lymphocytes were observed in the gut-associated lymphoid tissues. However, CD3+ cells were not apparent until day 12 and CD5+ cells were not detected until day 74 postpartum. No lymphocytes were detected in liver or bone marrow samples and no bronchus associated lymphoid tissues were observed. The pattern of development and the distribution of T- and B-cells in the lymphoid and immunohaematopoietic tissues were similar to those observed in eutherian mammals and in limited studies of other metatherians. However, the detection of apparently mature T- and B-cells in the thymus and gut-associated lymphoid tissues (GALT) at the same postnatal age highlights the need for a more substantial study of the development of GALT. This is, at present, limited by availability of marsupial-specific antibodies. PMID- 12892412 TI - Measurement of the linear dynamics of the descent of the bovine fetal testis. AB - Measurements were made on 86 male bovine fetuses collected from abattoirs in the vicinity of Sydney, Australia. The fetal body length was used to calculate the approximate day of gestational age (DGA); most fetuses were between 60 and 150 DGA. The distances from the caudal pole of the kidney (metanephros) to, respectively, the tip of the scrotum, the distal end of the testis and the internal ring of the inguinal canal were measured, as well as the dimensions of the testis and gubernaculum testis. Distances of (1) testis to inguinal canal, (2) inguinal canal to scrotum, (3) testis to scrotum and (4) gubernaculum to scrotum were calculated from these measurements, which were made on both left and right sides. The total length of the gubernaculum testis increased during transabdominal passage and during transinguinal passage of the testis. Furthermore, the gubernaculum appeared to maintain the testis at a relatively fixed distance from the scrotum during transabdominal passage so that the inguinal canal appeared to move towards the testis. The greatest distance between the testis and the tip of the scrotum was found during the transinguinal passage of the testis and was 2.8 cm for the left testis and 2.3 cm for the right. When located within the scrotum, each testis was still 1.6-1.7 cm from the tip of the scrotum, so the distance to be traversed was only 0.6-1.2 cm. Following passage of the testis through the inguinal canal, the gubernaculum became shorter and its distal tip was displaced toward the distal end of the scrotum. Traction by the gubernaculum could account for the final transposition of the testis from the external inguinal ring to the scrotum. Other factors involved in displacement of the testis include differential growth patterns as well as increases in the dimensions of the testis itself. PMID- 12892413 TI - Qualitative and quantitative determination of yohimbine in authentic yohimbe bark and in commercial aphrodisiacs by HPLC-UV-API/ MS methods. AB - The development and validation of a rapid qualitative and quantitative method based on an HPLC-UV-MS technique with atmospheric pressure chemical ionisation and electrospray ionisation for the analysis of yohimbine in a number of commercial aphrodisiac products is reported. HPLC with multiple-stage mass spectrometry experiments allowed the identification of the target compound and increased the selectivity of complex analyses such as those involved with multi botanical preparations. The precision and the robustness of the method were improved by the use of two internal standards: codeine for UV detection and deuterium-labelled yohimbine for MS detection. Twenty commercial aphrodisiac preparations were analysed and the amount of yohimbine measured and expressed as the maximal dose per day suggested on product labels ranged from 1.32 to 23.16 mg. PMID- 12892414 TI - Fractionation of grape tannins and analysis by matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry. AB - Polymeric tannins, extracted from grape berries (Gamay variety), were fractionated according to their mean degree of polymerisation (mDP) on a styrene divinylbenzene phase eluted with a gradient of methanol:chloroform. Increasing the percentage of methanol led to the solubilisation of higher molecular weight tannins. The mean mDP of each collected fraction was determined by acid-catalysed degradation in the presence of a nucleophilic reagent. The fractionation method produced a linear gradient of mDP varying between 1.84 and 19.34. The fractions were partially characterised by matrix assisted laser desorption/ionisation time of-flight mass spectrometry (MALDI-TOF/MS). The spectra showed a complex mixture of proanthocyanidins and galloylated proanthocyanidins up to 4000 amu. PMID- 12892415 TI - Isolation of high-quality RNA from white spruce tissue using a three-stage purification method and subsequent cloning of a transcript from the PR-10 gene family. AB - Isolation of PinmIII cDNA homologues from white spruce tissues required a rigorous RNA extraction protocol developed following assessment of three previously reported conifer RNA extraction protocols. Total RNA was extracted via several purification steps designed to minimize binding of phenolics to nucleic acids and was then subjected to caesium chloride ultra-centrifugation. This procedure produced consistently high-quality, intact RNA from both needles and roots with spectrophotometric ratios of approximately 2.0 for both 260/280 nm and 260/230 nm. Total RNA was obtained from the roots of cold-hardened white spruce seedlings for cDNA library construction. More than 2 million recombinant phage particles were generated from 5 microg of a poly(A)+RNA fraction, and ca. 1.3 million cDNA particles were amplified for storage. Approximately 500,000 primary recombinant clones were screened with an heterologous PinmIII cDNA sequence yielding a unique clone, picgl, that was very similar to members of the PR10 gene family. PMID- 12892416 TI - Chemiluminescence inhibition assay for folic acid using flow injection analysis. AB - A new flow injection method for the determination of folic acid is described. A fast oxidation reaction occurred when folic acid was mixed with potassium ferricyanide generating ferrocyanide which then inhibited the chemiluminescent reaction of ferricyanide and luminol in alkaline medium. The decrease of chemiluminescence intensity was correlated with the folic acid concentration in the range 0.1-21 microg/mL; the detection limit for the assay was 0.03 microg/mL (3sigma). A complete analysis of folic acid, including sampling and washing, could be performed within 2 min with a relative standard deviation of less than 4.0%. The proposed method has been applied successfully to the determination of folic acid in pharmaceutical preparations. PMID- 12892417 TI - Supercritical fluid extraction of naringin from the peel of Citrus paradisi. AB - The highest yield (14.4 g/kg) of naringin, the major flavonoid from the peel of Citrus paradisi L., that could be achieved by supercritical fluid extraction was obtained using supercritical carbon dioxide modified with 15% ethanol and fresh (rather than dried) peels at 95 bar and 58.6 degrees C. This yield is higher than that attained by the conventional technique of maceration, and close to those obtained by reflux and Soxhlet methods. Furthermore, supercritical fluid extraction consumes less solvent and provides a shorter extraction time than conventional extraction methods. PMID- 12892418 TI - A simple differential pulse polarographic method for the determination of thymoquinone in black seed oil. AB - A reliable and simple differential pulse polarographic method is described for the determination of thymoquinone in black seed oil. The polarographic behaviour of thymoquinone was examined in various buffer systems over the pH range 5.0 10.0. Thymoquinone is reduced in a single, reversible peak at the dropping mercury electrode. The differential pulse polarogram showed a distinct peak in Sorensen buffer:methanol (3:7, v/v; pH 8.5) at a peak potential of -0.095 V (vs. silver/silver chloride electrode), and a plot of peak height against concentration was found to be linear over the range 0.2-15.0 microg/mL (R = 0.9998). The limit of detection was calculated to be 0.054 microg/mL. The polarographic method has been applied to determine thymoquinone in two black seed oil preparations available on the Austrian pharmaceutical market. PMID- 12892419 TI - Chemical polymorphism of the essential oils from populations of Thymus caespititius grown on the islands Pico, Faial and Graciosa (Azores). AB - The compositions of the essential oils isolated from the aerial parts of 11 populations of Thymus caespititius collected during the flowering phase on Pico, Faial and Graciosa (Azores) were studied by GC and GC-MS. The monoterpene fraction was dominant in all the oils analysed (55-90%) and consisted mainly of oxygen-containing compounds (44-79%). Sesquiterpenes represented an important fraction of the oils from the populations grown on Graciosa (13-28%). In contrast, this fraction was rather small in the oils from the populations grown on Pico and Faial (6-11%). Despite this, oxygen-containing compounds (4-18%) were always dominant. Cluster analysis of all identified oil components grouped the oils into three main clusters that corresponded with their main components. The oils from the 11 populations studied showed a clear chemical polymorphism that, in some cases, was more evident among populations growing on the same island than among those from different islands. PMID- 12892420 TI - Monitoring seasonal variation in apple fruit volatile emissions in situ using solid-phase microextraction. AB - Emissions of volatiles from apple fruits (Malus domestica Borkh.) were monitored in situ over the course of a growing season (from early June to mid September) for two apple varieties, Golden Delicious and Maigold. Results indicate a characteristic time-course of volatile emissions as the sampling date was a statistically significant factor for nine of the 13 compounds considered. The amounts of volatiles collected were greatest early and late in the season. The temporal effect on emissions was generally much larger than the effect of variety, which was significant for only four of the 13 compounds considered. The possible sources of variation which are not explained by the statistical models are discussed, and it is considered that they are most likely related to differences in the emissions from individual fruits. PMID- 12892421 TI - Enantiomeric differentiation of bornyl acetate by 13C-NMR using a chiral lanthanide shift reagent. AB - The enantiomeric differentiation of bornyl acetate was carried out by 13C-NMR spectroscopy using a chiral lanthanide shift reagent. The technique was successfully applied to the determination of the enantiomer of bornyl acetate present in the essential oil of Inula graveolens. PMID- 12892422 TI - Seasonal variability of phytoplankton at Varna Bay (Black Sea). AB - Intrinsic fluorescence and SDS-PAGE analysis were employed to study the seasonal qualitative and quantitative changes of phytoplankton composition at Varna Bay (Black Sea). Variation in the maximum emission wavelength (lambda(max)) of the phytoplankton proteins (398 nm in the summer and 340 nm in the spring) was observed. In addition, a decrease in fluorescence intensity, and a shift in lambda(max) as a result of changes in phytoplankton protein stability, according to the season, was noted. Similarly, SDS-PAGE analysis showed different protein patterns for each season, for example in summertime the major protein constituents were of 14, 37, 48 and 70 kDa, while in the springtime the sizes ranged between 38 and 48 kDa. In general, higher carbohydrate and protein contents correlated with larger phytoplankton biomass found during the summer. The dominant species, the Bacillariophyceae and Dinophyceae, were found to be present in the water body in an alternate pattern. All of these changes could be accounted for by the adaptation of the organism to seasonal variations that modify the sea environment at Varna Bay. PMID- 12892423 TI - Lipids of the hexane extract from the roots of medicinal boraginaceous species. AB - The chemical compositions of hexane extracts of the lipid fraction of the roots of the medicinal Boraginaceous species Alkanna tinctoria, Onosma heterophylla, Macrotomia densiflora and Onosma hispidium are presented and their phytochemical relevance evaluated. The predominating fatty acids in all of the root lipids were stearic, palmitic, oleic, linoleic and gamma-linolenic acids, while the latter and stearidonic acid predominated in the seeds and leaves of various Boraginaceous species. The indigenous presence of methyl, ethyl and isopropyl esters of fatty acids, reported for the first time in the roots of higher plants, is considered to be of particular importance in the biosynthesis of fatty substances. The results suggest the use of fatty acids as chemotaxonomic markers for Boraginaceous species and the utilisation of Boraginaceous species as new commercial sources for fatty acids with valuable medicinal and nutritional properties. PMID- 12892424 TI - Preparative isolation of swainsonine from locoweed: extraction and purification procedures. AB - The trihydroxy indolizidine alkaloid swainsonine, a plant toxin with potent alpha mannosidase-inhibitory activity and chemotherapeutic potential, was isolated in gram quantities from locoweed (Astragalus lentiginosus). The key isolation and purification step was a continuous liquid/liquid extraction procedure using dichloromethane to extract a basified aqueous methanol solution obtained after isolation of the polar base fraction by ion-exchange. The concentration of swainsonine was increased from ca. 7% in the polar base material to 68% using the liquid/liquid extraction procedure. Pure swainsonine was then obtained by recrystallisation from ammonia-saturated chloroform or by sublimation. Small samples of swainsonine were also purified by formation of the chloroform-soluble methylboronate derivative, from which the alkaloid could be regenerated easily by hydrolysis. PMID- 12892425 TI - Seeing--by art and science. PMID- 12892426 TI - Do intersections serve as basic features in visual search? AB - The status of 'intersection' as a basic feature in visual search tasks has been controversial. Under some circumstances, a target possessing this attribute (eg a plus) 'pops out' of a display of distractors that lack the attribute (eg Ls). However, those cases may be artifacts of other features such as relative size or number of line terminators. We report two sets of experiments with stimuli intended to control for these factors. Search for the presence or absence of intersections is very inefficient with these stimuli. The results suggest that intersection should not be included among the list of salient features that support efficient search through visual displays. PMID- 12892427 TI - Visual context integration is not fully developed in 4-year-old children. AB - Long-range horizontal interactions supporting contour integration were found to be weaker in children than in adults (Kovacs et al, 1999 Proceedings of the National Academy of Sciences of the USA 96 12204-12209). In the present study, integration on a larger scale, between a target and its context was investigated. Contextual modulation of the percept of a local target can be directly measured in the case of geometric illusions. We compared the magnitude of a size contrast illusion (Ebbinghaus illusion or Titchener circles) in children and adults. 4 year-old children and adults performed 2AFC size comparisons between two target disks in the classical Ebbinghaus illusion display and in two other modified versions. We found that the magnitude of the illusion effect was significantly smaller in children than in adults. Our interpretation is that context integration is not fully developed in 4-year-old children. Closer-to-veridical size estimations by children demonstrate that the perception of the local target is less affected by stimulus context in their case. We suggest that immature cortical connectivity is behind the reduced contextual sensitivity in children. PMID- 12892428 TI - Is color an intrinsic property of object representation? AB - The role of color in object representation was examined by using a variation of the Stroop paradigm in which observers named the displayed colors of objects or words. In experiment 1, colors of color-diagnostic objects were manipulated to be either typical or atypical of the object (eg a yellow banana versus a purple banana). A Stroop-like effect was obtained, with faster color-naming times for the typical as compared to the atypical condition. In experiment 2, naming colors on words specifying these same color-diagnostic objects reversed this pattern, with the typical condition producing longer response times than the atypical condition. In experiment 3, a blocked condition design that used the same words and colors as experiment 2 produced the standard Stroop-like facilitation for the typical condition. These results indicate that color is an intrinsic property of an object's representation at multiple levels. In experiment 4, we examined the specific level(s) at which color-shape associations arise by following the tasks used in experiments 1 and 2 with a lexical-decision task in which some items were conceptually related to items shown during color naming (eg banana/monkey). Priming for these associates was observed following color naming of words, but not pictures, providing further evidence that the color-shape associations responsible for the differing effects obtained in experiments 1 and 2 are due to the automatic activation of color-shape associations at different levels of representation. PMID- 12892429 TI - Perceptual switching, eye movements, and the bus paradox. AB - According to a widely cited finding by Ellis and Stark (1978 Perception 7 575 581), the duration of eye fixations is longer at the instant of perceptual reversal of an ambiguous figure than before or after the reversal. However, long fixations are more likely to include samples of an independent random event than are short fixations. This sampling bias would produce the pattern of results also when no correlation exists between fixation duration and perceptual reversals. When an appropriate correction is applied to the measurement of fixation durations, the effect disappears. In fact, there are fewer actual button-presses during the long intervals than would be expected by chance. Moving-window analyses performed on eye-fixation data reveal that no unique eye event is associated with switching behaviour. However, several indicators, such as blink frequency, saccade frequency, and the direction of the saccade, are each differentially sensitive to perceptual and response-related aspects of the switching process. The time course of these indicators depicts switching behaviour as a process of cascaded stages. PMID- 12892430 TI - Timing accuracy in motion extrapolation: reverse effects of target size and visible extent of motion at low and high speeds. AB - By varying target size, speed, and extent of visible motion we examined the timing accuracy in motion extrapolation. Small or large targets (0.2 or 0.8 deg) moved at either 2.5, 5, or 10 deg s(-1) across a horizontal path (2.5 or 10 deg) and then vanished behind an occluder. Observers responded when they judged that the target had reached a randomly specified position between 0 and 12 deg. With higher speeds, the timing accuracy (the reverse of absolute error) was better for small than for large targets, and for long than for short visible extents. With low speed, these effects were reversed. In addition, while long visible extents yielded a greater accuracy at high than at low speeds, for short extents the accuracy was much better with the low speed. The findings suggest that, when extrapolating motion with targets and visible extents of different sizes, the visual system implements different scaling algorithms depending on target speed. At higher speeds, processing of visible and occluded motion is likely to share a common scaling mechanism based on velocity transposition. Reverse effects for target size and extent of visible motion at low and high speeds converge with the assumption of two distinct speed-tuned motion-processing mechanisms in human vision. PMID- 12892431 TI - Distance perception for points at equiconvergence and equidistance loci. AB - Subjects viewed luminescent targets placed at an equidistance (ED) locus or on a comparable equiconvergence locus, the Vieth-Muller circle (VMC). In darkness, subjects moved the index finger to the apparent distance of the visible target. This was repeated at leftward eccentricities of 0 degrees (straight ahead), 15 degrees, 22.5 degrees, 30 degrees, 37.5 degrees, and 45 degrees. Contrary to theoretical expectations, apparent distance to points on the VMC did not describe an equidistance locus, but diminished significantly with increasing eccentricity, whereas ED settings increased slightly but significantly. Binocular and monocular groups performed equivalently. Veridicality of results appears to be the outcome of an accommodation-convergence conflict with a dominant role for accommodation. PMID- 12892432 TI - A fair test of the effect of a shadow-incompatible luminance gradient on the simultaneous lightness contrast. AB - Shadow-compatibility of simultaneous lightness contrast is discussed by Alexander D Logvinenko and Paola Bressan, with examples claiming to provide a test of the hypothesis. PMID- 12892433 TI - Invalid cues impair auditory motion sensitivity. AB - Compelling lateral motion can be experienced when intensity differences between the two cars change over time. Whether our sensitivity to this dynamic interaural stimulation could be influenced by directional cues was the focus of the present study. On each trial, amplitude-modulated pure tones were presented either diotically (no-motion condition) or dichotically (motion condition), and participants indicated whether lateral motion was present or absent. Randomly across trials, the stimuli were preceded by a valid directional cue, an invalid directional cue, or no cue, while the motion to be detected was identical across these cue conditions. The data indicate that motion sensitivity was comparable in the valid-cue and no-cue conditions. Relative to each of those conditions, however, motion sensitivity was significantly lower in the invalid-cue condition, and motion was reported significantly less often. The results provide evidence that our sensitivity to dynamic interaural intensity differences can be significantly affected by a non-sensory factor, namely cue validity. PMID- 12892434 TI - Measuring and modeling real-time responses to music: the dynamics of tonality induction. AB - We examined a variety of real-time responses evoked by a single piece of music, the organ Duetto BWV 805 by J S Bach. The primary data came from a concurrent probe-tone method in which the probe tone is sounded continuously with the music. Listeners judged how well the probe tone fit with the music at each point in time. The process was repeated for all probe tones of the chromatic scale. A self organizing map (SOM) [Kohonen 1997 Self-organizing Maps (Berlin: Springer)] was used to represent the developing and changing sense of key reflected in these judgments. The SOM was trained on the probe-tone profiles for 24 major and minor keys (Krumhansl and Kessler 1982 Psychological Review 89 334-368). Projecting the concurrent probe-tone data onto the map showed changes both in the perceived keys and in their strengths. Two dynamic models of tonality induction were tested. Model 1 is based on pitch class distributions. Model 2 is based on the tone transition distributions; it tested the idea that the order of tones might provide additional information about tonality. Both models contained dynamic components for characterizing pitch strength and creating pitch memory representations. Both models produced results closely matching those of the concurrent probe-tone data. Finally real-time judgments of tension were measured. Tension correlated with distance away from the predominant key in the direction of keys built on the dominant and supertonic tones, and also correlated with dissonance. PMID- 12892435 TI - Mach bands change asymmetrically during solar eclipses. AB - Observations made during two partial eclipses of the Sun show that the Mach bands on shadows cast by the Sun disappear and reappear asymmetrically as an eclipse progresses. These changes can be explained as due to changes in the shape of the penumbras of shadows as the visible portion of the Sun forms crescents of different orientation. PMID- 12892436 TI - Junk science: appearances can be deceiving. PMID- 12892437 TI - Prehistory and the third molar. PMID- 12892438 TI - Tooth whitening. PMID- 12892439 TI - More about tooth whitening. PMID- 12892440 TI - Formation of mucogingival defects associated with intraoral and perioral piercing: case reports. AB - BACKGROUND: The authors provide clinical findings in five patients wearing oral jewelry to illustrate the risks of experiencing periodontal injury associated with body piercing involving intraoral and perioral sites. They also present a literature review of other adverse dental and medical consequences attributed to oral piercing. CASE DESCRIPTIONS: Five young adult patients with tongue and lip piercing sought dental care. Each patient exhibited some degree of gingival recession and mucogingival defects in proximity of their oral jewelry. Three of these patients had probing depths ranging from 5 to 8 millimeters in the affected areas. CLINICAL IMPLICATIONS: Intraoral and perioral jewelry may be associated with the development of significant mucogingival deformities. Because severe attachment loss can develop even when gingival recession is minimal, it is critical that patients with oral piercing routinely undergo comprehensive periodontal assessment. The authors urge clinicians to educate patients about the potential risks regarding the practice of oral piercing. PMID- 12892441 TI - Noncarious cervical lesions and abfractions: a re-evaluation. AB - BACKGROUND: The concept of abfraction is controversial. The authors present the fundamental basis of abfraction and review studies that describe the condition. TYPES OF STUDIES REVIEWED: The authors used data on masticatory forces, enamel and dentin properties, as well as stress studies related to abfraction, for background information. They also analyzed the genesis of the abfraction theory, experimental evidence, case presentations, clinical investigations and restorative studies. RESULTS: The theory of abfraction is based primarily on engineering analyses that demonstrate theoretical stress concentration at the cervical areas of teeth. While some recent stress studies support earlier findings, others have provided significant deviating information. Few controlled studies exist that demonstrate the relationship between occlusal loading and abfraction lesions. The role of occlusal loading in noncarious cervical lesions (as shown by clinical data) appears to be part of a multifactorial event that may not necessarily follow the proposed classic abfraction mechanism, and other mechanisms or factors may explain cervical restoration failure just as well. CLINICAL IMPLICATIONS: There is little direct evidence supporting the theory of abfraction as the primary factor in the formation of noncarious cervical lesions. Controlled clinical trials are necessary to elucidate more fully the etiology of those lesions. PMID- 12892442 TI - Isolation of an unusual fungus in treated dental unit waterlines. AB - BACKGROUND: Numerous organisms have been identified in dental unit waterlines, or DUWLs. Decontamination of DUWLs focuses on maintaining heterotrophic, mesophilic bacteria below 200 colony-forming units per milliliter as recommended by the ADA. METHODS: The authors conducted a study to test the efficacy of a continuous-use, stabilized chlorine dioxide proprietary compound to decrease the number of bacteria in DUWLs. The authors used three dental units with self-contained water systems to test the product and three similar units as controls. They aseptically collected water samples weekly according to recommended methods, plated the samples on R2A agar and incubated them for seven days. RESULTS: The authors isolated heterotrophic, mesophilic bacteria from treatment and control units for eight weeks. In the ninth week, the predominant isolates from one of the treatment units changed in appearance to small, dark, shiny colonies that the authors tentatively identified as fungal. The authors then isolated similar colonies from the source tap water and ultrasonic and handpiece lines. They added three additional dental units from the same clinic in the sixth week of the study and isolated similar fungal colonies from them after five weeks of treatment. The authors performed DNA sequencing with an automated sequencer and identified the organism Exophiala mesophila. CONCLUSIONS: The authors did not observe fungal isolates in the control units, which suggests that continuous waterline treatment may cause proliferation of a fungus present in small amounts in source water. CLINICAL IMPLICATIONS. The findings of this study indicate the need to monitor water quality regularly when treating waterlines with continuous-use chemical cleaners. PMID- 12892443 TI - The overexpression of cyclo-oxygenase-2 in chronic periodontitis. AB - BACKGROUND: The objective of this prospective cross-sectional study was to determine if cyclo-oxygenase-2, or COX-2, is overexpressed in the inflamed gingival tissue of patients diagnosed as having moderate-to-severe chronic periodontitis, or CP. METHODS: The authors evaluated clinical measures, crevicular fluid and gingival biopsy specimens from patients with moderate or severe CP (n = 16) and from healthy volunteers (n = 8). Patients were diagnosed as having CP based on clinical attachment loss, or CAL, of at least 5 millimeters at two sites in each quadrant and on evidence of alveolar bone loss as assessed from standard periapical or bite-wing radiographs. Healthy patients exhibited no sites with CAL of more than 2 mm and no evidence of alveolar bone loss. The authors used standard techniques to perform biochemical measures. RESULTS: Levels of interleukin-1 beta, or IL-1beta, in crevicular fluid were more than doubled in the CP group (P < .05). The amounts of COX-2 mRNA and protein also were elevated in gingival tissues from subjects with CP compared with those from healthy subjects. To gain further mechanistic insights, the authors conducted in vitro studies. The results showed that lipopolysaccharide and tumor necrosis factor alpha, or TNF-alpha, induced COX-2 in macrophages, while IL-1beta and TNF-alpha induced COX-2 in oral epithelial cells. CONCLUSIONS: Taken together, these results suggest that levels of COX-2 in gingivae reflect clinical measures of periodontitis and gingival inflammation. CLINICAL IMPLICATIONS: The discovery of increased levels of COX-2 in inflamed gingival tissue suggests that COX-2 represents a pharmacological target for the prevention or treatment of CP. PMID- 12892444 TI - Injection pain: comparison of three mandibular block techniques and modulation by nitrous oxide:oxygen. AB - BACKGROUND: The authors conducted this study to compare the pain on injection of the three mandibular block techniques and to determine the effect that the delivery of nitrous oxide:oxygen, or N2O:O2, had on this pain. METHODS: Based on sample size calculation, the authors randomly assigned 60 subjects to receive either local anesthetic alone or local anesthetic with N2O:O2 titrated to effect. Each subject received two mandibular block injections bilaterally from three possible pairings: Gow-Gates/standard block, standard block/Vazirani-Akinosi or Gow-Gates/ Vazirani-Akinosi. Subjects scored their pain using a 100-millimeter visual analog scale immediately after each injection. The authors analyzed data using analysis of variance, Student t tests and multiple regression analyses. RESULTS: There were no significant differences in pain on injection among the three injection techniques. The subjects in the group that received N2O:O2 and the local anesthetic demonstrated a statistically significant reduction in pain on injection compared with subjects in the group that received the local anesthetic only (P < .05). When N2O:O2 was used, there was a statistically significant decrease in pain with the first injection (P < .0005), an effect not seen with the second injection. CONCLUSIONS: There was no significant difference in pain among the three mandibular block techniques. N2O:O2 can reduce pain on the first injection given, but this effect is not seen subsequently. CLINICAL IMPLICATIONS: The decision to select one of the three mandibular block techniques should be based on factors other than pain on injection. N2O:O2 reduces pain on the first injection only. PMID- 12892445 TI - Maxillary odontogenic keratocyst: a common and serious clinical misdiagnosis. AB - BACKGROUND: The authors conducted this study to determine the most common location of odontogenic keratocysts, or OKCs, in the maxilla and the implication of this for dental practice. METHODS: The authors reviewed all cases diagnosed as an OKC by the Oral and Maxillofacial Pathology Diagnostic Laboratory at University of Florida College of Dentistry from July 1, 1993, to June 30, 2001. They determined the location for each cyst and recorded relevant clinical information such as age and sex. In addition, the authors noted the practitioners' clinical diagnoses for the cases in the most common location in the maxilla and compared them with the final diagnosis. RESULTS: A total of 398 OKCs occurred in 393 patients. Of the 398 cysts, 266 (66.8 percent) were in the mandible, and 132 (33.2 percent) were in the maxilla. The most common location for OKCs was the third molar and ramus area of the mandible, with 137 cysts. The canine region of the maxilla, with 54 cysts, was the second most common location. Submitting clinicians mentioned OKC as a diagnostic possibility in only 31.5 percent of the maxillary canine region cases. CONCLUSION: The most common maxillary location for OKCs is the canine region where they commonly are mistaken for an apical inflammatory lesion or lateral periodontal cyst. Accurate diagnosis is essential for proper patient therapy and follow-up. CLINICAL IMPLICATION: A diagnosis based solely on clinical information can be problematic. In many cases, histologic examination of surgically removed tissue is necessary to establish a definitive diagnosis. That is particularly true for cases that do not resolve when treated by conservative endodontic therapy. PMID- 12892446 TI - A technique for direct placement of transitional anterior resin veneers. PMID- 12892447 TI - Avoiding complications in local anesthesia induction: anatomical considerations. AB - BACKGROUND: Dentists administer thousands of local anesthetic injections every day with few reports of serious complications. However, misjudging the anatomy involved during local anesthetic administration can result not only in inadequate or incomplete anesthesia, but in other complications such as paresthesia, bleeding or hematoma formation, or in serious systemic complications. OVERVIEW: The authors discuss anatomical considerations that dentists should keep in mind when administering local anesthetic injections. In particular, they discuss ways in which dentists can minimize the risk of nerve or vascular injury, as well as systemic complications. CONCLUSIONS AND CLINICAL IMPLICATIONS: Even the most experienced practitioner can benefit from a periodic review of the anatomy associated with local anesthesia. This article offers dentists the opportunity to consider needle placement with regard to location of nerves, blood vessels and glands, and to review injection protocols that can minimize the risk of complications. PMID- 12892448 TI - Antibiotic prophylaxis for dental patients with total joint replacements. AB - BACKGROUND AND OVERVIEW: In 1997, the American Dental Association and the American Academy of Orthopaedic Surgeons convened an expert panel of dentists, orthopaedic surgeons and infectious disease specialists and published their first Advisory Statement on Antibiotic Prophylaxis for Dental Patients with Prosthetic Joints. This represented the first time that national health organizations had gone on record on this topic. This 2003 advisory statement is the first periodic update of the 1997 statement. In addition, the organizations have created a new patient handout (included at the end of the statement) that dentists may share with their patients. The 1997 Advisory Statement has been well-used by dentists and orthopaedic surgeons. Following their standard protocols for periodic review of existing advisory statements, the ADA and AAOS and their expert consultants recently reviewed the 1997 statement. CONCLUSIONS AND CLINICAL IMPLICATIONS: The 2003 statement includes some modifications of the classification of patients at potential risk and of the incidence stratification of bacteremic dental procedures, but no changes in terms of suggested antibiotics and antibiotic regimens. The statement concludes that antibiotic prophylaxis is not indicated for dental patients with pins, plates or screws, nor is it routinely indicated for most dental patients with total joint replacements. However, it is advisable to consider premedication in a small number of patients who may be at potential increased risk of experiencing hematogenous total joint infection. PMID- 12892449 TI - Antiseptic antimicrobial hand washes. PMID- 12892450 TI - Hand hygiene. PMID- 12892451 TI - Are shareholders employees of professional corporations? PMID- 12892452 TI - Ensuring retention for crowns and fixed prostheses. AB - Crowns and fixed prostheses are well-proven, accepted and routinely used restorations. However, they occasionally come loose from tooth preparations. Many things can cause these failures. In this article, I have discussed the following reasons for lack of adequate retention of crowns and fixed prostheses: inadequate tooth preparation; too much trust in dentin bonding agents and lack of adequate tooth buildup; tooth preparations that lack irregularities; improper selection of cements; and lack of postoperative occlusal adjustment. PMID- 12892453 TI - The value of new patients. AB - Make no mistake: dental practices are businesses. Dentists as CEOs need to focus on key performance indicators and statistical tracking in order to understand what is happening in the practice over its life span. Average production per new patient is a key factor to be tracked because it is an essential driver of the financial performance of any dental practice. The health of this factor will have a dramatic effect on the financial well-being of the dentist, and on the timing and character of his or her retirement. At the same time, it is essential to balance financial factors with exceptional quality of care for every patient. Without both, neither will be successful in the long-term. PMID- 12892454 TI - For the dental patient. Dental sealants. Protecting your teeth. PMID- 12892455 TI - Influences of thoracic duct blockage on early enteral nutrition for patients who underwent esophageal cancer surgery. AB - OBJECTIVES: We have previously reported the beneficial effects of immediate enteral nutrition (EN) after esophageal cancer surgery. This randomized control study was conducted to determine whether immediate EN is beneficial or not for patients whose thoracic ducts were ligated, as well as those whose thoracic ducts were preserved. PATIENTS AND METHODS: Thirty-nine patients who underwent radical resection of the esophageal cancer entered this trial. After stratifying into two groups--patients whose thoracic ducts were preserved [D(+)] and those whose thoracic ducts were ligated [D(-)], they were randomly divided into two groups- the patients who received early EN and those who received parenteral nutrition (PN) followed by delayed enteral feeding. Thus, the number of patients in the D(+)-EN group, D(+)-PN group, D(-)-EN group and D(-)-PN group were 13, 12, 7 and 7, respectively. The mortality and morbidity rates, and several blood chemistries were compared between the EN groups and the PN groups. RESULTS: Total lymphocyte count showed a significant early increase and serum c-reactive protein (CRP) was significantly decreased in the D(+)-EN group compared to the D(+)-PN group. However those differences were not observed between the D(-) groups. Serum total bilirubin was significantly decreased in the both EN groups compared to the PN groups. The mortality and morbidity rates were not different between the EN group and the PN group in the D(+) patients and also in the D(-) patients. CONCLUSIONS: Patients whose thoracic ducts were ligated did not obtain any other benefit from early enteral feeding except for bilirubin metabolism. Early enteral feeding is not recommended for patients whose thoracic ducts are ligated during radical resection of a cancer in the thoracic esophagus. PMID- 12892457 TI - Sequential bypass using the right gastroepiploic artery for coronary artery bypass grafting. AB - OBJECTIVE: Since 1989, we have applied the right gastroepiploic artery (RGEA) as a third arterial conduit for coronary artery bypass grafting (CABG) and started to use sequential RGEA in 1992. We evaluated the feasibility and efficacy of sequential RGEA grafting in CABG. METHODS: From December 1990 to January 2000, 46 patients underwent CABG with sequential RGEA. There were 42 male and 4 female patients with a mean age of 59 +/- 8.1 years. Mean postoperative follow-up period was 70 months. RESULTS: The mean number of anastomoses was 3.7 per patient. Mean luminal diameter of the RGEA was 2.2 +/- 0.4 mm by preoperative angiography and 2.3 +/- 0.6 mm by intraoperative measurement. Patency of the sequential RGEA was 92%; proximal anastomosis 100%, distal anastomosis 86% (p = 0.01). The 5-year actuarial survival and cardiac event-free rate were 91% and 93%, respectively. CONCLUSIONS: Sequential bypass using the RGEA is feasible, with excellent early and long-term results. The indication for sequential RGEA, however, needs careful anatomical consideration of both the luminal diameter of the RGEA and proximal stenosis of the target coronary arteries. PMID- 12892456 TI - Immunohistochemical study of the expression of drug-resistant proteins in large cell neuroendocrine carcinoma of the lung. AB - OBJECTIVES: In 1999, the World Health Organization categorized pulmonary large cell neuroendocrine carcinoma as a variant of large cell carcinoma. However, an optimal treatment for large cell neuroendocrine carcinoma has not been established yet. Recently, multimodality therapy combining both surgery and adjuvant chemotherapy has been reported as a useful treatment for large cell neuroendocrine carcinoma, but the effect of chemotherapy on it has not yet been fully investigated. Thus, we evaluated immunohistochemical data of the expression of drug-resistant proteins in large cell neuroendocrine carcinoma. METHODS: We identified 10 large cell neuroendocrine carcinomas (1.2%) out of 850 primary lung cancers that had been surgically resected. We examined the immunohistochemical staining of three drug-resistant proteins, namely, P-glycoprotein, metallothionein and glutathione S-transferase-pi to compare large cell neuroendocrine carcinoma with other histological types of lung cancer. RESULTS: The mean tumor cell positivity rates for P-glycoprotein, metallothionein and glutathione S-transferase-pi in large cell neuroendocrine carcinoma were 0%, 2.4 +/- 3.6% and 35.0 +/- 37.5%, respectively. The positivity rates for P glycoprotein and glutathione S-transferase-pi were significantly lower than those in adenocarcinoma (P = 0.0003, P = 0.0009). The positivity rate for glutathione S transferase-pi was also lower than that in squamous cell carcinoma (P = 0.0387). These drug-resistant proteins showed similar expression pattern in both large cell neuroendocrine carcinoma and small cell carcinoma except glutathione S transferase-pi. CONCLUSION: Immunohistochemical expression of drug-resistant proteins in large cell neuroendocrine carcinoma was lower than that in adenocarcinoma and squamous cell carcinoma, and differences exist in drug resistance between large cell neuroendocrine carcinoma and small cell carcinoma. PMID- 12892458 TI - Late results of mitral valve repair for mitral regurgitation. AB - OBJECTIVE: This study was undertaken to evaluate the long-term results of mitral valve repair for mitral regurgitation. METHODS: Between 1991 and 2000, 301 patients with mitral regurgitation underwent mitral valve repair. There were 167 men and 134 women whose mean age was 56 +/- 14 years. The patients were comprised of 7 patients in Carpentier's type I, 277 patients in type II, and 17 patients in type III. Chordal replacement with expanded polytetrafluoroethylene sutures had been prospectively applied to repair the anterior mitral leaflet prolapse. Ring annuloplasty was performed in 230 patients (76%). The follow-up was complete and mean follow-up was 67 +/- 33 months, for a cumulative follow-up of 1,624 patient years. RESULTS: There were 5 hospital deaths and 11 late deaths (2 cardiac and 9 noncardiac). All survivors except those with stroke were in the New York Heart Association (NYHA) functional class I or II. At 10 years, the actuarial survival was 90 +/- 3%, the freedom from embolism was 86 +/- 4%, the freedom from reoperation was 96 +/- 2%, and the freedom from valve-related events was 77 +/- 4%. At 10 years, the freedom from reoperation in the patients with anterior leaflet prolapse was 90 +/- 5%. CONCLUSIONS: Mitral valve repair is feasible in most patients with mitral regurgitation and is associated with low mortality and low rates of valve related events. Chordal replacement with expanded polytetrafluoroethylene sutures is effective, safe, and durable at long-term follow-up for patients with anterior leaflet prolapse. PMID- 12892459 TI - Surgery for pulmonary metastases from colorectal cancer: the importance of prethoracotomy serum carcinoembryonic antigen as an indicator of prognosis. AB - OBJECTIVE: Several investigators have analyzed prognostic factors of surgical treatment for pulmonary metastases from colorectal cancer, but the results remain inconclusive. This study was performed to determine the prognostic implications of the prethoracotomy serum level of carcinoembryonic antigen (CEA) in relation to the postthoracotomy recurrent pattern among patients with this disease. METHODS: A retrospective analysis of prognostic factors was undertaken in 100 patients who had consecutively undergone initial surgical resection for pulmonary metastases of colorectal origin. RESULTS: The overall 3- and 5-year survival rates were 62.2% and 49.4%, respectively. Univariate analysis revealed that the prethoracotomy serum CEA level and operative curability were strongly associated with prognosis, while in multivariate analysis, only the prethoracotomy serum CEA level was a significant prognostic indicator. Patients with a high level of prethoracotomy serum CEA more frequently exhibited recurrence in extrathoracic sites, especially in the brain. CONCLUSION: Before thoracotomy for pulmonary metastases from colorectal cancer, the serum CEA level was the most useful prognostic factor. Patients with elevated serum CEA level should undergo a careful prethoracotomy systemic survey and postthoracotomy follow-up for extrathoracic metastases, in particular brain metastases, and an appropriate combined therapeutic modality should be considered. PMID- 12892460 TI - Independent predictive value of the overall number of metastatic N1 and N2 stations in lung cancer. AB - OBJECTIVE: The number of metastatic N2 stations is a known prognostic factor in patients with non-small-cell lung cancer (NSCLC). However, involvement of N1 stations as well as that of N2 stations seems to be important in the prognosis of these patients. We therefore attempt to clarify the significance of the total number of metastatic stations in pathologic N1 and N2 NSCLCs. METHODS: Patients with either pathologic N1 (n = 51) or N2 (n = 96) NSCLC who had undergone major pulmonary resection with complete mediastinal dissection were included in this retrospective study. All positive nodes were characterized by location according to the TNM classification system. The hilar station was included with the N2 stations. RESULTS: The total number of metastatic stations in patients with N2 disease ranged from 1 to 8 (average 2.5), whereas that in patients with N1 disease ranged from 1 to 3 (average 1.2). The incidence of multiple-station metastasis (> or = 3 metastatic stations) among N2 patients (35%) was significantly higher than that among N1 patients (2%) (p < 0.001). Multivariate analysis of survival showed pathologic N1 status (relative risk = 0.443, p = 0.013) and < or = 2 metastatic stations (relative risk = 0.515, p = 0.020) to be significant and independent prognostic factors. Age, sex, cell type, resected lobe, and pathological T status were statistically insignificant determinates of survival. CONCLUSIONS: The total number of metastatic stations (< or = 2 vs > or = 3) is an independent prognostic indicator in patients with completely resected pathologic N1 or N2 NSCLC. The number of metastatic stations will be useful as a stratification factor in prospective clinical trials of these patients. PMID- 12892461 TI - The limitation of staged repair in the surgical management of congenital complex heart anomalies with aortic arch obstruction. AB - OBJECTIVE: Severe aortic arch obstruction including an interrupted aortic arch in congenital complex heart anomalies remains a challenge in surgical management. METHODS: Treatment and outcomes in 75 consecutive patients who underwent an aortic arch repair as the first step of the staged repair protocol between 1975 and 2000 were reviewed. Their ages at repair ranged from 1 day to 8.5 months. RESULTS: Cross-sectional postoperative follow-up data were available in all the patients. The follow-up period ranged from 0 to 27.6 years (mean: 7.3 +/- 7.3 years). There were 20 postoperative hospital deaths (27%) and 7 late deaths. The Kaplan-Meier estimate of survival was 81.3% +/- 4.5% at 1 month, 68.0% +/- 5.4% at 1 year, 65.0% +/- 5.5% at 5 years, 63.1% +/- 5.7% at 10 years, 63.1% +/- 5.7% at 20 years. By Cox regression analysis, body weight of 2.5 kg or less is the only independent determinant of postoperative mortality (p = 0.04, multivariable odds ratio: 2.50, [95% confidence interval: 1.02-6.1]). The aortic arch morphology, the primary cardiac lesion, or date of operation did not reach a statistically significant level to show correlation with mortality. Reintervention to reconstruct the aortic arch was performed at 9 occasions in 8 of the 55 patients who survived the primary operation (14.5%). The Kaplan-Meier estimate of the reintervention-free rate was 91.3% +/- 4.2% at 5 years, 85.5% +/- 5.6% at 10 years, 75.6% +/- 8.2% at 20 years. Using multivariable Cox regression analysis, interrupted aortic arch (versus aortic coarctation) was the only independent predictor of a shorter time to reintervention (p = 0.001, multivariable odds ratio: 16.1, [95% confidence interval: 3.2-80.2]). CONCLUSIONS: The staged repair protocol was associated with significant limitations in patient survival and with the development of recurrent aortic arch obstruction. Thus, a primary repair protocol may serve as an alternate approach, especially in patients with low weight or with an interrupted aortic arch. PMID- 12892462 TI - Use of a larynyngeal mask airway during aortic valve replacement. AB - We report the elective use of a laryngeal mask airway during cardiac surgery for congenital tracheal stenosis. A 53-year-old woman with severe aortic valve stenosis was scheduled for aortic valve replacement. During anesthesia induction, the anesthesiologists attempted conventional intubation but failed. Fiberoptic tracheal examination and computed tomography showed a tracheal stenosis with 5 mm minimal diameter. A laryngeal mask airway was used at the patient's rescheduled surgery. The laryngeal mask airway use did not lead any surgical complication. We concluded that the laryngeal mask airway may be considered as an alternative to conventional intubation in patients with tracheal stenosis. PMID- 12892463 TI - A unique collision of epithelioid hemangioendothelioma and bronchioloalveolar carcinoma in a 54-year-old Japanese woman. AB - We present a unique collision of epithelioid hemangioendothelioma and bronchioloalveolar carcinoma. A 54-year-old Japanese woman seen for a small nodular shadow on chest X-ray in December 1989 was found in chest computed tomography to have multiple small nodules in bilateral lung fields. At thoracotomy through a median sternotomy, 9 nodules from 0.3 cm to 1.2 cm in diameter detected were all resected. The pathological diagnosis was epithelioid hemangioendothelioma of the lung. A focus of bronchioloalveolar carcinoma 1.4 cm in diameter existed adjacent to the focus of epithelioid hemangioendothelioma in the lingular portion of the left lung. The initial surgical margin was not clear, and a complete lingulectomy was conducted through a left lateral thoracotomy. The patient is currently alive with no evidence of recurrence at 13 years after the first appearance. PMID- 12892464 TI - Surgical treatment for cervical aortic arch with aneurysm formation. AB - Cervical aortic arch is an unusual malformation. Cervical aortic arch with aneurysm formation is very rare. We report a case of cervical aortic arch associated with a saccular aneurysm in a 59-year-old Japanese man. The aneurysm protruded caudally and was located between the left common carotid and left subclavian arteries. Cardiopulmonary bypass and deep hypothermic circulatory arrest was applied as adjunct methods. A Dacron graft was sutured just distal to the left common carotid artery, with the patient in the Trendelenburg position. The proximal site was left open while oxygen-saturated venous blood was supplied in a retrograde manner to perfuse the lower body during occlusion of the descending aorta. Distal anastomosis to the descending aorta was performed during rewarming. The left subclavian artery was reconstructed by using a branch of the graft. This procedure is simple and useful for distal arch operations, especially in patients with Haughton D type aneurysms. PMID- 12892465 TI - Ruptured pseudoaneurysm of the aorta with encapsulated mediastinal abscess after coronary artery bypass grafting. AB - Rupture of pseudoaneurysm of the aorta with chronic active mediastinitis after cardiac surgery is a catastrophic complication. We report a case of one-stage operation of urgent repair for ruptured pseudoaneurysm under deep hypothermic circulatory arrest, debridement of mediastinum, and omental transfer, with an uneventful postoperative course. PMID- 12892466 TI - Successful replacement of aortic root with valve-sparing technique and proximal arch in a patient with myelodysplastic syndrome. AB - Cardiac surgery in a patient with myelodysplastic syndrome (MDS) increases the risk of bleeding and infection. Here we report a case of a 70-year-old man with MDS who underwent successful replacement of the aortic root with the valve sparing technique and proximal arch for aneurysmal dilatation from the aortic root to ascending aorta with moderate aortic valve regurgitation. Perioperatively, a transfusion of red blood cells and an infusion of a grannulocyte colony-stimulating factor were required for his serious erythrocytopenia and leukocytopenia. Bleeding tendency was so severe that re exploration to control postoperative surgical bleeding was performed and a large amount of blood cells were transfused. There was no infection on the postoperative course. Perioperative management for cardiac surgery in patients with MDS must be carefully programmed by a co-operative team consisting of cardiovascular surgeons and hematologists. PMID- 12892467 TI - Long-term survival in adult mediastinal neuroblastoma. AB - Neuroblastoma rarely occurs in adults, in whom prognosis is poor. Here we report on the effective use of multimodal therapy to achieve long-term survival in adult mediastinal neuroblastoma. In a 33-year-old male with severe pain radiating from the left hypochondrium to the back area, no abnormal shadow was detected on first examination; the shadow appeared in chest X-ray only 10 months later when the severe pain recurred. The patient was then referred to our department for further examination. Chest X-rays and chest CT scans revealed a posterior mediastinal mass beside the thoracic vertebrae. Diagnosis was confirmed after surgical resection, which was followed by outpatient adjuvant therapy through radiation and chemotherapy. The patient has survived 8 years and 8 months without recurrence or metastasis. PMID- 12892468 TI - Solitary cardiac metastasis of rectal adenocarcinoma. AB - Solitary cardiac metastasis is rarely recognized. We report a case of solitary cardiac metastasis from a rectal adenocarcinoma that was manifested as superior vena cava (SVC) syndrome. Invasion of the deep cardiac structures was so severe that only its palliative resection and right atrial reconstruction were performed using a cardiopulmonary bypass to release the SVC obstruction. The postoperative course was uneventful and the patient was discharged home without any complications. Adjuvant chemotherapy could not be performed because of the renal dysfunction. Eleven months later, the patient died from multiple metastasis of adenocarcinoma. Even though the surgery was not curative, it might be effective for release from symptoms and for elongating life expectancy. PMID- 12892469 TI - Primary tissue failure of the Freestyle stentless aortic root bioprosthesis. AB - We present a case of early tissue failure of the Medtronic Freestyle stentless bioprosthesis used in the aortic position which led to reoperation 27 months after implantation. Almost one half of the left coronary cusp of the prosthetic valve had torn away along the annulus. The tear started from the left-non coronary commissure and reached the midpoint of the base of the left coronary cusp. The prosthetic valve was easily excised and replaced with a new mechanical valve. There was no sign of calcification or vegetation in the excised valve. The cause of the valve dysfunction was thought to be primary tissue failure. Long term function of the Freestyle stentless valve should be carefully followed up. PMID- 12892470 TI - Techno-economic evaluation of producing ethanol from softwood: comparison of SSF and SHF and identification of bottlenecks. AB - The aim of the study was to evaluate, from a technical and economic standpoint, the enzymatic processes involved in the production of fuel ethanol from softwood. Two base case configurations, one based on simultaneous saccharification and fermentation (SSF) and one based on separate hydrolysis and fermentation (SHF), were evaluated and compared. The process conditions selected were based mainly on laboratory data, and the processes were simulated by use of Aspen plus. The capital costs were estimated using the Icarus Process Evaluator. The ethanol production costs for the SSF and SHF base cases were 4.81 and 5.32 SEK/L or 0.57 and 0.63 USD/L (1 USD = 8.5SEK), respectively. The main reason for SSF being lower was that the capital cost was lower and the overall ethanol yield was higher. A major drawback of the SSF process is the problem with recirculation of yeast following the SSF step. Major economic improvements in both SSF and SHF could be achieved by increasing the income from the solid fuel coproduct. This is done by lowering the energy consumption in the process through running the enzymatic hydrolysis or the SSF step at a higher substrate concentration and by recycling the process streams. Running SSF with use of 8% rather than 5% nonsoluble solid material would result in a 19% decrease in production cost. If after distillation 60% of the stillage stream was recycled back to the SSF step, the production cost would be reduced by 14%. The cumulative effect of these various improvements was found to result in a production cost of 3.58 SEK/L (0.42 USD/L) for the SSF process. PMID- 12892471 TI - Hypothermic preservation of hepatocytes. AB - This paper presents a review of recent research on the hypothermic storage of hepatocytes. The first focus is on the diversity of methodologies currently employed in this area. The cell damage caused by hypothermic preservation and its possible mechanism are then investigated on both morphological and molecular biology. Later, the gene expressions on a mRNA level or enzyme level after hypothermic preservation are further discussed. Finally, the improvement of hypothermic storage by preconditioning, such as by increasing temperature, is explored. PMID- 12892472 TI - Kinetic modeling of the autotrophic growth of Pavlova lutheri: study of the combined influence of light and temperature. AB - The optimization and control of biochemical processes require the previous establishment of mathematical models that can describe the effect of process variables on their actual kinetics. Environmental temperature is a modulating factor to which the algal cells respond continuously by adjusting their rates of cellular reactions, their nutritional requirements, and, consequently, their biomass composition. Light intensity is an exhaustible resource, indispensable to autotrophic organisms. The effects of light intensity and temperature on growth of the microalga Pavlova lutheri, which have hardly been considered to date in a simultaneous fashion, were experimentally assessed using a factorial experimental design; in this way, the effects of each variable independently and their interactions could be quantified, using maximum biomass (X(max)) or maximum specific growth rate (mu(max)) as objective functions. The preliminary results produced indicated that light intensity plays a more important role on mu(max) than temperature; in the case of X(max), both temperature and, to a lesser extent, light intensity do apparently play a role. The highest values of X(max) were associated with low temperatures and high light intensities; a similar behavior could be observed for mu(max) concerning light intensity, although the dependency on temperature did not seem to be as important. A more complex mechanistic model was then postulated, incorporating light and temperature as input variables, which was successfully fitted to the experimental data generated during batch cultivation of P. lutheri. PMID- 12892473 TI - Metabolic control analysis of xylose catabolism in Aspergillus. AB - A kinetic model for xylose catabolism in Aspergillus is proposed. From a thermodynamic analysis it was found that the intermediate xylitol will accumulate during xylose catabolism. Use of the kinetic model allowed metabolic control analysis (MCA) of the xylose catabolic pathway to be carried out, and flux control was shown to be dependent on the metabolite levels. Due to thermodynamic constraints, flux control may reside at the first step in the pathway, i.e., at the xylose reductase, even when the intracellular xylitol concentration is high. On the basis of the kinetic analysis, the general dogma specifying that flux control often resides at the step following an intermediate present at high concentrations was, therefore, shown not to hold. The intracellular xylitol concentration was measured in batch cultivations of two different strains of Aspergillus niger and two different strains of Aspergillus nidulans grown on media containing xylose, and a concentration up to 30 mM was found. Applying MCA showed that the first polyol dehydrogenase (XDH) in the catabolic pathway of xylose exerted the main flux control in the two strains of A. nidulans and A. niger NW324, but the flux control was exerted mainly at the first enzyme of the pathway (XR) of A. niger NW 296. PMID- 12892474 TI - An adaptive strategy for single- and multi-cluster gene assignment. AB - Strict assignment of genes to one class, dimensionality reduction, a priori specification of the number of classes, the need for a training set, nonunique solution, and complex learning mechanisms are some of the inadequacies of current clustering algorithms. Existing algorithms cluster genes on the basis of high positive correlations between their expression patterns. However, genes with strong negative correlations can also have similar functions and are most likely to have a role in the same pathways. To address some of these issues, we propose the adaptive centroid algorithm (ACA), which employs an analysis of variance (ANOVA)-based performance criterion. The ACA also uses Euclidian distances, the center-of-mass principle for heterogeneously distributed mass elements, and the given data set to give unique solutions. The proposed approach involves three stages. In the first stage a two-way ANOVA of the gene expression matrix is performed. The two factors in the ANOVA are gene expression and experimental condition. The residual mean squared error (MSE) from the ANOVA is used as a performance criterion in the ACA. Finally, correlated clusters are found based on the Pearson correlation coefficients. To validate the proposed approach, a two way ANOVA is again performed on the discovered clusters. The results from this last step indicate that MSEs of the clusters are significantly lower compared to that of the fibroblast-serum gene expression matrix. The ACA is employed in this study for single- as well as multi-cluster gene assignments. PMID- 12892475 TI - Chiral resolution function with immobilized food proteins. AB - We confirmed that an NAD(P)+-dependent secondary alcohol dehydrogenase (NAD(P)-E) can be easily and effectively isolated from pea, soybean, and wheat proteins immobilized with calcium alginate gel (IPP, ISP, and IWP, respectively). The estimated molecular mass of NAD(P)-E is 138.7 kDa, and the concentrations of NAD(P)-E in solution are 36.2 (IPP), 53.9 (ISP), and 93.7 (IWP) microg/mL. The NAD(P)-E oxidizes only (R)-isomers highly enantioselectively; thus, greater than 99% ee(s) of (S)-isomers can be obtained from corresponding rac-aryl methyl carbinols (1, 2a-6a, and 2b-7b). The amount of food protein needed for 1 g of substrate (B/S ratio) is approximately 20. Thus, in comparison to current biocatalysts, certain food proteins can serve as asymmetric reagent bases, providing easily obtained, low-cost natural catalysts with stereoselectivity, regioselectivity, and substrate specificity that work under mild conditions for asymmetric synthesis of organic compounds. Moreover, this "fourth" function of food may help build a sustainable society by synthesizing optically active secondary alcohols in an environmentally friendly manner. PMID- 12892476 TI - Bioaugmentation strategies for remediating mixed chemical effluents. AB - Operationally exhausted metal working fluids are chemically mixed, produced in large quantities (400 000 tonnes year in the U.K.), and potentially environmentally toxic. It is essential to develop more reliable and economical approaches for their disposal. We investigated the effectiveness of a defined bacterial consortium, constructed specifically for treating metal-working fluid (MWF), and contrasted its performance to that of undefined inocula from activated sludge. Construction of the consortium was based on knowledge of the diversity of bacterial communities that naturally colonize MWF and determination of their catabolic abilities and tolerance to the chemical constituents. Chemical analysis of the inoculated MWF bioreactor revealed that, after 100 h at 28 degrees C, the defined inoculum reduced the pollution load by over 80% from an initial chemical oxygen demand of approximately 48 000 mg L(-)(1). The inocula performance was approximately 50% more effective than that of the undefined microbial community from the activated sludge. Furthermore, the performance of the constructed consortium was more reproducible than that of an undefined community, an essential feature for bioaugmentation treatment of industrial wastes. PMID- 12892478 TI - Conjugation of penicillin acylase with the reactive copolymer of N isopropylacrylamide: a step toward a thermosensitive industrial biocatalyst. AB - Conjugation of penicillin acylase (PA) to poly-N-isopropylacrylamide (polyNIPAM) was studied as a way to prepare a thermosensitive biocatalyst for industrial applications to antibiotic synthesis. Condensation of PA with the copolymer of NIPAM containing active ester groups resulted in higher coupling yields of the enzyme (37%) compared to its chemical modification and copolymerization with the monomer (9% coupling yield) at the same NIPAM:enzyme weight ratio of ca. 35. A 10 fold increase of the enzyme loading on the copolymer resulted in 24% coupling yield and increased by 4-fold the specific PA activity of the conjugate. Two molecular forms of the conjugate were found by gel filtration on Sepharose CL 4B: the lower molecular weight fraction of ca. 10(6) and, presumably, cross-linked protein-polymer aggregates of MW > 10(7). Michaelis constant for 5-nitro-3 phenylacetamidobenzoic acid hydrolysis by the PA conjugate (20 microM) was found to be slightly higher than that of the free enzyme (12 microM), and evaluation of V(max) testifies to the high catalytic efficiency of the conjugated enzyme. PolyNIPAM-cross-linked PA retained its capacity to synthesize cephalexin from d phenylglycin amide and 7-aminodeacetoxycephalosporanic acid. The synthesis hydrolysis ratios of free and polyNIPAM-cross-linked enzyme in cephalexin synthesis were 7.46 and 7.49, respectively. Thus, diffusional limitation, which is a problem in the industrial production of beta-lactam antibiotics, can be successfully eliminated by cross-linking penicillin acylase to a smart polymer (i.e., polyNIPAM). PMID- 12892477 TI - Porcine pancreatic alpha-amylase hydrolysis of native starch granules as a function of granule surface area. AB - Porcine pancreatic alpha-amylase activity on native starch granules is more accurately described as a function of surface area of the granules rather than of substrate concentration. The apparent K(m) of alpha-amylolysis of native starch from potato, maize, and rice expressed as a function of substrate concentration was largest for potato with a single value of V(max). However, the ratio of the slope of a Lineweaver-Burk plot to that of rice for enzymatic hydrolysis of native potato and maize starch were 7.78 and 2.58, respectively, which were very close to the ratio of surface area per mass of the two starch granules to that of rice. Therefore, the reciprocal of initial velocity was a linear function of the reciprocal of surface area for each starch granule. Surface area was calculated assuming the starch granules were spherical. The values obtained by this calculation were in good agreement with the value obtained by the photomicrographic method. By comparing enzymatic digestion of native maize granules to that of rice granules, it was concluded that the presence of pores in maize granules appeared to significantly affect overall rate of digestion after sufficient reaction time, but not at the very initial stage of hydrolysis. PMID- 12892479 TI - Kinetics of whole serum and prepurified IgG digestion by pepsin for F(ab')2 manufacture. AB - An alternative route for the production of polyclonal F(ab')(2) fragments that might be adopted for the facile preparation of antivenoms is assessed in this work. The method involves the digestion of whole serum by free pepsin, which results in reduction of the number of processing steps commonly in use, because it avoids the initial purification of IgG's prior to their proteolytic cleavage by the enzyme. Digestion kinetics of whole serum and caprylic acid prepurified IgG using free pepsin were monitored with SDS-PAGE followed by densitometric analysis and antigen binding activity assay of the digested samples. It was observed that with equal units of pepsin activity, caprylic acid prepurified IgG was digested more rapidly than whole serum but that the overall retention of antigen binding activity was significantly greater in the latter case. The estimated first-order digestion rate parameters were 11.8 and 4.42 microM min( )(1) for pure IgG and whole serum, respectively. The K(m) value obtained for whole serum digestion was 33 microM and that for pure IgG digestion was 43.5 microM. Calibration with undigested whole serum and pure IgG samples of known concentrations was performed using SDS-PAGE followed by image analysis. A linear relationship was observed between the protein concentration and the respective band intensity within the range of concentrations investigated (0.63-31.2 microM IgG concentration). This technique proved to be relatively rapid, reproducible, and more precise than size-exclusion chromatography as a result of its F(ab')(2)/IgG resolving power. Staining and destaining protocols were reproduced in terms of staining and destaining times, volumes added, and compositions. Furthermore, all digestion experiments were performed in duplicate sets to monitor the extent of variation of the digestion kinetic parameters measured by this method. The results obtained from this technique confirm and quantify previous observations that pepsin digestion of whole serum is slower and easier to control than digestion of pure IgG and results in higher recovery of antigenic binding activity. PMID- 12892480 TI - Experimental and theoretical analysis of tubular membrane aeration for Mammalian cell bioreactors. AB - A combination of experimental and theoretical approaches was used to characterize the dynamics of oxygen transfer in a membrane-aerated bioreactor. Pressure profiles along the length of the membrane at varying entrance and exit pressures were determined by actual experimental measurements, unlike most previous studies that have relied solely on theoretical descriptions of the pressure profile in the tubing. The mass transfer coefficient, k(L)a, was also determined under these conditions and was found to be essentially independent of tubing exit pressure. Measurement of the tubing pressure profile coupled with estimation of k(L)a allowed for computation of the oxygen transfer rate (OTR) along the length of the tubing. A mathematical model that incorporated friction pressure loss and losses due to tubing bending was developed to describe the pressure and hence OTR characteristics of membrane-aerated systems. The applicability of the model was verified by testing it on experimentally measured pressure data, and in all cases the model accurately described experimental data. When tubing properties are known, the mathematical model presented in this study allows for a priori estimation of OTR profiles along the length of the tubing. This information is vital for optimal design and scale-up of membrane-aerated bioreactors for mammalian cell culture. PMID- 12892481 TI - Productivity improvement in xanthan gum fermentation using multiple substrate optimization. AB - A novel and more comprehensive formulation of the optimal control problem that reflects the operational requirements of a typical industrial fermentation has been proposed in this work. This formulation has been applied to a fed-batch bioreactor with three control variables, i.e., feed rates of carbon source, nitrogen source, and an oxygen source, to result in a 148.7% increase in product formation. Xanthan gum production using Xanthomonas campestris has been used as the model system for this optimization study, and the liquid-phase oxygen supply strategy has been used to supply oxygen to the fermentation. The formulated optimization problem has several constraints associated with it due to the nature of the system. A robust stochastic technique, differential evolution, has been used to solve this challenging optimization problem. The infinite dimensional optimization problem has been approximated to a finite dimensional one by control vector parametrization. The state constraints that are path constraints have been addressed by using penalty functions and by integrating them over the total duration to ensure a feasible solution. End point constraints on final working volume of the reactor and on the final residual concentrations of carbon and nitrogen sources have been included in the problem formulation. Further, the toxicity of the oxygen source, H(2)O(2), has been addressed by imposing a constraint on its maximum usable concentration. In addition, the initial volume of the bioreactor contents and feed concentrations have been handled as decision variables, which has enabled a well-grounded choice for their values from the optimization procedure; adhoc values are normally used in the industry. All results obtained by simulation have been validated experimentally with good agreements between experimental and simulated values. PMID- 12892482 TI - Effect of shear stress on intrinsic CHO culture state and glycosylation of recombinant tissue-type plasminogen activator protein. AB - Shear stress in suspension culture was investigated as a possible manipulative parameter for the control of glycosylation of the recombinant tissue-type plasminogen activator protein (r-tPA) produced by recombinant Chinese hamster ovary (CHO) cell culture, grown in protein-free media. Resulting fractions of partially glycosylated, Type II, and fully glycosylated, Type I, r-tPA protein were monitored as a direct function of the shear characteristics of the culture environment. The shear-induced response of CHO culture to levels of low shear stress, where exponential growth was not obtained, and to higher levels of shear stress, which resulted in extensive cell death, were examined through manipulation of the bioreactor stirring velocity. Both apparent and intrinsic cell growth, metabolite consumption, byproduct and r-tPA production, and r-tPA glycosylation, from a variable site-occupancy standpoint, were monitored throughout. Kinetic analyses revealed a shear-stress-induced alteration of cellular homeostasis resulting in a nonlinear dependency of metabolic yield coefficients and an intrinsic cell lysis kinetic constant on shear stress. Damaging levels of shear stress were used to investigate the shear dependence of cell death and lysis, as well as the effects on the intrinsic growth rate of the culture. Kinetic models were also developed on the basis of the intrinsic state of the culture and compared to traditional models. Total r-tPA production was maximized under moderate shear conditions, as was the viable CHO cell density of the culture. However, Type II r-tPA production and the fraction of Type II glycoform production ratio was maximized under damaging levels of shear stress. Analyses of biomass production yield coefficients coupled with a plug-flow reactor model of glycan addition in the endoplasmic reticulum (ER) were used to propose an overall mechanism of decreased r-tPA protein site-occupancy glycosylation with increasing shear stress. Decreased residence time of r-tPA in the ER as a result of increased protein synthesis related to shear protection mechanisms is proposed to limit contact of site Asn184 with the membrane-bound oligosaccharyltransferase enzyme in the ER. PMID- 12892483 TI - Xylitol production from sugarcane bagasse hydrolyzate in fluidized bed reactor. Effect of air flowrate. AB - Cells of Candida guilliermondii immobilized onto porous glass spheres were cultured batchwise in a fluidized bed bioreactor for xylitol production from sugarcane bagasse hemicellulose hydrolyzate. An aeration rate of only 25 mL/min ensured minimum yields of xylose consumption (0.60) and biomass production (0.14 g(DM)/g(Xyl)), as well as maximum xylitol yield (0.54 g(Xyt)/g(Xyl)) and ratio of immobilized to total cells (0.83). These results suggest that cell metabolism, although slow because of oxygen limitation, was mainly addressed to xylitol production. A progressive increase in the aeration rate up to 140 mL/min accelerated both xylose consumption (from 0.36 to 0.78 g(Xyl)/L.h) and xylitol formation (from 0.19 to 0.28 g(Xyt)/L.h) but caused the fraction of immobilized to total cells and the xylitol yield to decrease up to 0.22 and 0.36 g(Xyt)/g(Xyl), respectively. The highest xylitol concentration (17.0 g(Xyt)/L) was obtained at 70 mL/min, but the specific xylitol productivity and the xylitol yield were 43% and 22% lower than the corresponding values obtained at the lowest air flowrate, respectively. The concentrations of consumed substrates and formed products were used in material balances to evaluate the xylose fractions consumed by C. guilliermondii for xylitol production, complete oxidation through the hexose monophosphate shunt, and cell growth. The experimental data collected at variable oxygen level allowed estimating a P/O ratio of 1.35 mol(ATP)/mol(O) and overall ATP requirements for biomass growth and maintenance of 3.4 mol(ATP)/C mol(DM). PMID- 12892484 TI - Modeling stability of photoheterotrophic continuous cultures in photobioreactors. AB - Continuous cultures of the purple non-sulfur bacterium Rhodospirillum rubrum were grown in a cylindrical photobioreactor in photoheterotrophic conditions, using acetate as carbon source. A new kinetic and stoichiometric knowledge model was developed, and its ability to simulate experimental results obtained under varying incident light fluxes and residence times is discussed. The model accurately predicts the stable, unstable, or oscillating behavior observed for the reactor productivity. In particular, the values of residence time corresponding to a subcritical bifurcation with a typical hysteresis effect are calculated and analyzed. The robustness of the proposed model allows the engineering operating domain of the photobioreactor function to be set and offers a promising tool for the design and control of such photoheterotrophic processes. PMID- 12892485 TI - Modeling the kinetics of enzymic reactions in mainly solid reaction mixtures. AB - There is currently considerable interest in using mainly solid reaction mixtures for enzymic catalysis. In these reactions starting materials dissolve into, and product materials crystalize out of, a small amount of liquid phase in which the catalytic reaction occurs. An initial mathematical model for mass transfer effects in such systems is constructed using some physically reasonable approximations. The model equations are solved numerically to determine how the reactant concentrations vary with time and position. To evaluate the extent to which mass transfer limits the overall rate of product formation, an effectiveness factor is defined as the ratio of the observed total reaction rate to the total reaction rate in the reaction limited limit. As expected, the value of the effectiveness factor in steady state is strongly dependent on the Thiele modulus. However, it is also observed that the effectiveness factor can vary widely as a result of changes in the other dimensionless groups characterizing the system. For example, there are situations with Thiele modulus equal to unity in which the value of the effectiveness factor varies between approximately 0.1 and 0.8 as the other parameters are varied in physically reasonable ranges. Analytical asymptotic solutions that provide good approximations to the numerically calculated results in various physically important limiting cases are also presented. PMID- 12892486 TI - Oxidation of ABTS by silicate-immobilized cytochrome c in nonaqueous solutions. AB - Cytochrome c can be readily adsorbed onto mesoporous silicates at high loadings of up to 10 mmol g(-)(1) of silicate. The adsorbed protein retains its peroxidative activity, with no diffusional limitations being observed. The protein can be adsorbed onto the external surface of the silicate or, provided that the pore diameter is sufficiently large, into the channels. In aqueous buffer, the catalytic activity of the adsorbed protein (for the oxidation of ABTS) decreased with increasing temperature, with the decrease being less marked for cytochrome c held within the silicate channels. Similar results were obtained in 95% methanol. Analysis of kinetic data showed that significant increases in k(cat)/K(M) occurred in methanol, ethanol, and formamide, with slight decreases occurring in 1-methoxy-2-propanol. The observed increases were primarily a result of substantial increases in k(cat), while the results in 1-methoxy-2-propanol can be ascribed to increases in K(M). Resonance Raman spectroscopy indicated that the structure of the heme environment of the adsorbed protein was essentially unchanged, in aqueous buffer and in the nonaqueous solvents, methanol, 1-methoxy 2-propanol, and ethanol. In addition, Raman spectra of the lyophilized protein indicated that there were no apparent changes in the heme structure. PMID- 12892487 TI - Modeling and analysis of biooxidation of gold bearing pyrite-arsenopyrite concentrates by Thiobacillus ferrooxidans. AB - The results of modeling the biooxidation of a mixed sulfidic concentrate by Thiobacillus ferrooxidans is reported here. A kinetic model, which accounts for the dissolution of sulfide matrix due to both bacterial attachment onto the mineral surface and indirect leaching, has been proposed. A comprehensive system approach is employed for modeling the complex biooxidation process by (a) the decomposition of the complete system into several subsystems, (b) modeling individual systems, and (c) integrating the subsystems model in a final system model. The model for subsystems was developed by writing mass balance equations for the different species involved. The bacterial balance accounts for its growth, both on solid substrate and in solution, and for the attachment to and detachment from the surface. The kinetic parameters of the model were determined by designing the experiments in such a manner that only one subsystem was operational. This model was tested in both laboratory scale batch and continuous biooxidation processes. The model predictions agreed with the experimental data reasonably well. A further analysis of the model was carried out to predict the conditions for efficient biooxidation. Studies on the effect of residence time and pulp density on steady-state behavior showed that there is a critical residence time and pulp density below which washout conditions occur. Operation at pulp densities lower than 5% and residence times lower than 72 h was found unfavorable for efficient leaching. PMID- 12892488 TI - Lipase-catalyzed esterification of conjugated linoleic acid with sorbitol: a kinetic study. AB - The kinetics of esterification of conjugated linoleic acid (CLA) with sorbitol in acetone was investigated. An immobilized lipase from Candida antarctica (Chirazyme L-2) was used as the biocatalyst. A 2(2) x 3 factorial design was employed to find an experimental region in which one obtains a high rate of formation of the diester product. Best results were obtained at 10 degrees C using a CLA to sorbitol molar ratio of 5 and a biocatalyst loading of 150 mg/mL of acetone. Under these conditions, in 72 h one obtains a nearly quantitative yield (ca. 98%) of the diester of sorbitol with CLA. To minimize formation of products with degrees of esterification greater than two, the reaction should be carried out at 10 degrees C. A kinetic model developed using the King-Altman method was employed to fit the data. Use of the steady-state approximation for the monoester and an assumption that the concentration of sorbitol was constant and equal to its solubility limit permit one to minimize the number of parameters necessary to model the reaction network. Nonlinear regression analysis based on either two or three parameters provides very good fits of the multiresponse data in the presence or absence of triesters, respectively. PMID- 12892489 TI - Design and optimization of hot-filling pasteurization conditions: Cupuacu (Theobroma grandiflorum) fruit pulp case study. AB - Cupuacu (Theobroma grandiflorum) is an Amazonian tropical fruit with a great economic potential. Pasteurization, by a hot-filling technique, was suggested for the preservation of this fruit pulp at room temperature. The process was implemented with local communities in Brazil. The process was modeled, and a computer program was written in Turbo Pascal. The relative importance among the pasteurization process variables (initial product temperature, heating rate, holding temperature and time, container volume and shape, cooling medium type and temperature) on the microbial target and quality was investigated, by performing simulations according to a screening factorial design. Afterward, simulations of the different processing conditions were carried out. The holding temperature (T(F)) and time (t(hold)) affected pasteurization value (P), and the container volume (V) influenced largely the quality parameters. The process was optimized for retail (1 L) and industrial (100 L) size containers, by maximizing volume average quality in terms of color lightness and sensory "fresh notes" and minimizing volume average total color difference and sensory "cooked notes". Equivalent processes were designed and simulated (P(91)( degrees )(C) = 4.6 min on Alicyclobacillus acidoterrestris spores) and final quality (color, flavor, and aroma attributes) was evaluated. Color was slightly affected by the pasteurization processes, and few differences were observed between the six equivalent treatments designed (T(F) between 80 and 97 degrees C). T(F) >/= 91 degrees C minimized "cooked notes" and maximized "fresh notes" of cupuacu pulp aroma and flavor for 1 L container. Concerning the 100 L size, the "cooked notes" development can be minimized with T(F) >/= 91 degrees C, but overall the quality was greatly degraded as a result of the long cooling times. A more efficient method to speed up the cooling phase was recommended, especially for the industrial size of containers. PMID- 12892490 TI - Analysis of the phase behavior of the aqueous poly(ethylene glycol)-Ficoll system. AB - The PEG-Ficoll polymer phase system is one that has been overlooked in the past for biotechnology applications because of the stability of its emulsions. However, new applications, such as emulsion coating of cells, are appearing that rely on this very property. Ficoll is highly polydisperse and multimodal with three distinct Ficoll peaks in gel permeation chromatography. As a result, the transition between one-phase and two-phase systems is blurred and the binodials obtained through turbidometric titration and tie-line analysis differ significantly. Moreover, since the three Ficoll peaks partition differently, tie line analysis cannot be described by a simple model of the aqueous two-phase system. A simple modification to the model allowed for excellent fit, and this modification may prove well-suited for the many practical cases where aqueous two phase systems fail to display parallel tie-lines as implicitly assumed in the simpler model. PMID- 12892491 TI - Food-grade chemical and biological agents permeabilize red beet hairy roots, assisting the release of betalaines. AB - Hairy root cultures of red beet, Beta vulgaris L., were permeabilized under the functions of food-grade chemical and biological agents cetyl trimethylammonium bromide (CTAB), Triton X-100, Tween-80, Lactobacillus helveticus, Saccharomyces cereviseae, and Candida utilis, as well as cell fractions of L. helveticus, for the recovery of betalaines with or without oxygen stress. Tween-80 (0.15%), Triton X-100 (0.2%), and CTAB (0.05%), in combination with oxygen stress, released 45%, 70%, and 90% pigment into the medium, respectively, with significantly lesser levels in agitated cultures receiving similar treatments. The release was rapid (1 h) in CTAB treatment with a much slower release in Tween 80. CTAB (0.002%) was found to be also useful in effluxing betalaines (80%) from hairy roots grown in a bubble column reactor. Viability of permeabilized hairy roots, tested on agar medium, was not affected by any level of CTAB treatment and was significantly retarded at higher levels of Triton X-100 and Tween-80. An altogether new approach of pigment release using biological agents such as live cells of food-grade microbes was used where C. utilis, L. helveticus, and S. cereviseae released 60%, 85%, and 54% betalaines, respectively, in 24 h, though lower level treatments also released similar levels of pigment by 48 h. Dried whole cell powder of L. helveticus, its total insoluble carbohydrate, and free lipid fractions released 10%, 0%, and 85% pigment, respectively. An extended study with a bubble column reactor using the free lipid fraction of L. helveticus showed 50% and 84% pigment release in 8 and 12 h, respectively, exhibiting good viability when plated on agar medium. Even in the bioreactor, replenishment of medium 8 h after treatment with free lipid of L. helveticus allowed regrowth of hairy roots. The high level of pigment release recorded here, using CTAB or lipid of L. helveticus, appears useful for developing processes for in situ recovery of betalaines. The live microbes, applicable only for batch cultures, are expected to impart improved sensory/nutraceutical effects to the recovered pigment and hence may add value to the product receiving the red beet pigment thus produced. PMID- 12892492 TI - Use of a diffusion model for mono- and bicomponent anion-exchange of two isoenzymes of glucoamylase from Aspergillus niger in a fixed bed. AB - The adsorption of glucoamylases I and II (GA I and GA II, respectively) from Aspergillus niger on the anion exchanger DEAE-Toyopearl 650 was studied in fixed bed experiments, and the effect of temperature, flowrate, inlet concentration, bed length, and particle size on the process was characterized. The anion exchanger showed a higher adsorption capacity for the more active isoenzyme GA I in all experimental conditions studied. A mathematical model accounting for external and pore diffusion and nonlinear equilibrium isotherm (for GA I) was used to fit the experimental breakthrough curves, showing very accurate fittings in all of the operating conditions. The values of the pore diffusion coefficient at 15, 20, and 25 degrees C were, respectively, 1.25 x 10(-)(11), 1.46 x 10( )(11) and 1.83 x 10(-)(11) (for GA I) and 1.82 x 10(-)(11), 2.44 x 10(-)(11) and 2.73 x 10(-)(11) (for GA II) m(2)/s. Bicomponent adsorption experiments showed no significant interference effects between GA I and GA II, and so the mathematical model was again used to fit these experiments, yielding very satisfactory results. PMID- 12892493 TI - Purification and characterization of a novel thermo-alkali-stable catalase from Thermus brockianus. AB - A novel thermo-alkali-stable catalase from Thermus brockianus was purified and characterized. The protein was purified from a T. brockianus cell extract in a three-step procedure that resulted in 65-fold purification to a specific activity of 5300 U/mg. The enzyme consisted of four identical subunits of 42.5 kDa as determined by SDS-PAGE and a total molecular mass measured by gel filtration of 178 kDa. The catalase was active over a temperature range from 30 to 94 degrees C and a pH range from 6 to 10, with optimum activity occurring at 90 degrees C and pH 8. At pH 8, the enzyme was extremely stable at elevated temperatures with half lives of 330 h at 80 degrees C and 3 h at 90 degrees C. The enzyme also demonstrated excellent stability at 70 degrees C and alkaline pH with measured half-lives of 510 h and 360 h at pHs of 9 and 10, respectively. The enzyme had an unusual pyridine hemochrome spectrum and appears to utilize eight molecules of heme c per tetramer rather than protoheme IX present in the majority of catalases studied to date. The absorption spectrum suggested that the heme iron of the catalase was in a 6-coordinate low spin state rather than the typical 5 coordinate high spin state. A K(m) of 35.5 mM and a V(max) of 20.3 mM/min.mg protein for hydrogen peroxide was measured, and the enzyme was not inhibited by hydrogen peroxide at concentrations up to 450 mM. The enzyme was strongly inhibited by cyanide and the traditional catalase inhibitor 3-amino-1,2,4 triazole. The enzyme also showed no peroxidase activity to peroxidase substrates o-dianisidine and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid), a trait of typical monofunctional catalases. However, unlike traditional monofunctional catalases, the T. brockianus catalase was easily reduced by dithionite, a characteristic of catalase-peroxidases. The above properties indicate that this catalase has potential for applications in industrial bleaching processes to remove residual hydrogen peroxide from process streams. PMID- 12892494 TI - Development of rigid bidisperse porous microspheres for high-speed protein chromatography. AB - Development of a high-performance stationary phase is an essential demand for high-speed separation of proteins by liquid chromatography. Based on a novel porogenic mode, that is, using superfine granules of calcium carbonate as solid porogen and a mixture of cyclohexanol and dodecanol as liquid porogen, a rigid spherical biporous poly(glycidyl methacrylate-co-ethylene dimethacrylate) matrix has been prepared by radical suspension-polymerization. The epoxide groups of the matrix were modified with diethylamine to afford the ionizable weak base 1-N,N diethylamino-2-hydeoxypropy functionalities that are required for ion exchange chromatography. Results from scanning electron microscopy and mercury intrusion porosimetry measurements revealed that the matrix contained two families of pores, that is, micropores (10-90 nm) and macropores (180-4000 nm). Furthermore, the biporous medium possesses specific surface area as high as 91.3 m(2)/g. Because of the presence of the macropores that provided convective flow channels for the mobile phase, the dynamic adsorption capacity was found to be as high as 54.6 mg/g wet bead at 300 cm/h, approximately 63.2% of its static capacity. In addition, the column efficiency and dynamic binding capacity decreased only slightly with mobile-phase flow rate in the range of 300-3000 cm/h. These properties made the packed bed with the bidisperse porous matrix suitable for high-speed protein chromatography. PMID- 12892495 TI - Catalysis of protein folding by an immobilized small-molecule dithiol. AB - The isomerization of non-native disulfide bonds often limits the rate of protein folding. Small-molecule dithiols can catalyze this process. Here, a symmetric trithiol, tris(2-mercaptoacetamidoethyl)amine, is designed on the basis of criteria known to be important for efficient catalysis of oxidative protein folding. The trithiol is synthesized and attached to two distinct solid supports via one of its three sulfhydryl groups. The resulting immobilized dithiol has an apparent disulfide E degrees ' = -208 mV, which is close to that of protein disulfide isomerase (E degrees ' = -180 mV). Incubation of the dithiol immobilized on a TentaGel resin with a protein containing non-native disulfide bonds produced only a 2-fold increase in native protein. This dithiol appeared to be inaccessible to protein. In contrast, incubation of the dithiol immobilized on styrene-glycidyl methacrylate microspheres with the non-native protein produced a 17-fold increase in native protein. This increase was 1.5-fold greater than that of a monothiol immobilized on the microspheres. Thus, the choice of both the solid support and thiol can affect catalysis of protein folding. The use of dithiol-decorated microspheres is an effective new strategy for preparative protein folding in vitro. PMID- 12892496 TI - Quantifying process tradeoffs in the operation of chromatographic sequences. AB - A method for the rapid representation of key process tradeoffs that need to be made during the analysis of chromatographic sequences has been proposed. It involves the construction of fractionation and maximum purification factor versus yield diagrams, which can be completed easily on the basis of chromatographic data. The output of the framework developed reflects the degree of tradeoff between levels of yield and purity and provides a fast and precise prediction of the sample fraction collection strategy needed to meet a desired process specification. The usefulness of this approach for the purposes of product purification and contaminant removal in a single chromatographic step has been successfully demonstrated in an earlier paper and it is now extended by application to a chromatographic sequence: the separation of a hypothetical three component protein system by hydrophobic interaction chromatography (HIC) followed by size exclusion chromatography (SEC). The HIC operation has a strong impact upon the subsequent SEC step. The studies show how the analysis of performance in such a chromatographic sequence can be carried out easily and in a straightforward fashion using the fractionation diagram approach. The methodology proposed serves as a useful tool for identifying the process tradeoffs that must be made during operation of a sequence of chromatographic steps and indicates the impact on further processing of the cut-point decisions that are made. PMID- 12892497 TI - Biological reduction of nitric oxide in aqueous Fe(II)EDTA solutions. AB - The reduction of nitric oxide (NO) in aqueous solutions of Fe(II)EDTA is one of the core processes in BioDeNOx, an integrated physicochemical and biological technique for NO(x)() removal from industrial flue gases. NO reduction in aqueous solutions of Fe(II)EDTA (20-25 mM, pH 7.2 +/- 0.2) was investigated in batch experiments at 55 degrees C. Reduction of NO to N(2) was found to be biologically catalyzed with nitrous oxide (N(2)O) as an intermediate. Various sludges from full-scale denitrifying and anaerobic reactors were capable to catalyze NO reduction under thermophilic conditions. The NO reduction rate was not affected by the presence of ethanol or acetate. EDTA-chelated Fe(II) was found to be a suitable electron donor for the biological reduction of nitric oxide to N(2), with the concomitant formation of Fe(III)EDTA. In the presence of ethanol, EDTA chelated Fe(III) was reduced to Fe(II)EDTA. This study strongly indicates that redox cycling of FeEDTA plays an important role in the biological denitrification process within the BioDeNOx concept. PMID- 12892498 TI - Imaging polarimetry for high throughput chiral screening. AB - Imaging polarimetry was demonstrated as a highly parallel method of determining optical rotation of biochemical samples. The imaging polarimeter utilized a bright, uniform light source wavelength-filtered to near the sodium D line, a sample array flanked by inlet and analyzing polarizers, and a CCD camera fitted with an equal-perspective telecentric lens. The prototype apparatus was demonstrated to have an optical resolution better than 0.08 degrees. The potential for high throughput screening was demonstrated by imaging chiral solutions in 1536-well microtiter plates and by real-time monitoring of 30 simultaneous chiral enzymatic reactions. Improvements in polarizer and CCD technology may broadly expand the technique's applicability to fields such as directed evolution and combinatorial chemistry, where screening throughput is currently limiting for chiral applications. PMID- 12892499 TI - Monitoring and controlling the dissolved oxygen (DO) concentration within the high aspect ratio vessel (HARV). AB - A probe-type oxygen sensor was developed utilizing a radioluminescent (RL)-based light source and a ruthenium-based sensing chemistry for monitoring the dissolved oxygen (DO) concentration in a modified version of the NASA-designed high aspect ratio vessel (HARV), a batch rotating wall vessel. This sensor provided the means to monitor the DO concentration in the HARV without influencing the flow pattern, thereby retaining the low shear HARV environment conducive to the formation of 3 dimensional cell aggregates. This sensor lost significant signal as a result of exposure to the first three autoclave cycles, but only minimal change in signal was observed following exposure to subsequent autoclave cycles. A new calibration model requiring only one fitted parameter was developed that accurately fit data over the entire range from 0% to 100% oxygen saturation. The ability for DO concentration control within the vessel was demonstrated by using this sensor to monitor the DO concentration inside the HARV. PMID- 12892500 TI - Comparison of morphological characteristics of Streptomyces natalensis by image analysis and focused beam reflectance measurement. AB - A morphological interpretation is presented for data collected during growth of a filamentous organism, using a focused beam reflectance measurement (FBRM) system. The morphology of the organism was also obtained using conventional semiautomatic image analysis to support the interpretation of the FBRM data. The model organism employed is the filamentous soil-borne actinomycete Streptomyces natalensis, which produces the antifungal agent pimaricin. The organism was cultivated both in shake flasks and in a bench-scale stirred tank bioreactor. It was found that FBRM could be used to track changes in the morphology of the organism throughout the course of its growth on both scales. These changes were highlighted using both the median chord length and length-weighted mean chord length obtained from the chord length distribution measured with the FBRM probe. The ability of the FBRM probe to respond to changes in both the size and morphology of mycelial aggregates was supported by standard image analysis parameters, including equivalent diameter, convex area, and compactness. PMID- 12892501 TI - Measuring adsorption of a hydrophobic probe with a surface plasmon resonance sensor to monitor conformational changes in immobilized proteins. AB - Conformational changes of proteins immobilized on solid matrices were observed by measuring the adsorption of Triton X-100 (TX), a nonionic detergent, as a hydrophobic probe with BIACORE, a biosensor that utilizes the phenomenon of surface plasmon resonance (SPR). Two kinds of proteins, alpha-glucosidase and lysozyme, were covalently attached to dextran matrices on the sensor surface in the flow cell and then exposed to various concentrations of TX solution. We measured SPR signal changes derived from adsorption of TX to the immobilized proteins and calculated the monolayer adsorption capacity using the Brunauer Emmett-Teller (BET) equation. The results demonstrated that monolayer adsorption capacity is proportional to the amount of immobilized proteins. Further, the unfolding process of immobilized proteins on the sensor surface induced by guanidine hydrochloride was investigated by monitoring SPR signal increases due to the adsorption of TX to the exposed hydrophobic region of the protein. Results strongly suggested that the increase in the SPR signal reflected the formation of the agglutinative unfolded state. We expect our measuring method using the SPR sensor and TX adsorption will be a novel tool to provide conformational information regarding various proteins on solid matrices. PMID- 12892502 TI - Potential of on-line CIMS for bioprocess monitoring. AB - Chemical-ionization mass spectrometry (CIMS) using flow reactors is an emerging method for on-line monitoring of trace concentrations of organic compounds in the gas phase. In this study, a flow-reactor CIMS instrument, employing the H(3)O(+) cation as the ionizing reagent, was used to simultaneously monitor several volatile metabolic products as they are released into the headspace during bacterial growth in a bioreactor. Production of acetaldehyde, ethanol, acetone, butanol, acetoin, diacetyl, and isoprene by Bacillus subtilis is reported. Ion signal intensities were related to solution-phase concentrations using empirical calibrations and, in the case of isoprene, were compared with simultaneous gas chromatography measurements. Identification of volatile and semivolatile metabolites is discussed. Flow-reactor CIMS techniques should be useful for bioprocess monitoring applications because of their ability to sensitively and simultaneously monitor many volatile metabolites on-line. PMID- 12892503 TI - Production of tripeptide from gelatin using collagenase-immobilized porous hollow fiber membrane. AB - Tripeptide was produced during the permeation of a gelatin solution through the pore of a collagenase-immobilized porous hollow-fiber membrane. Gelatin was obtained via hydrolysis of fish collagen. First, an epoxy-group-containing monomer was graft-polymerized onto an electron-beam-irradiated porous hollow fiber membrane. Second, the 2-hydroxyethylamino group was introduced into the epoxy group to bind collagenase on the basis of electrostatic interaction. Third, adsorbed collagenase was cross-linked with glutaraldehyde to prevent leakage of the enzyme. Gelatin solution (10-50 g/L) was forced to permeate across the collagenase-immobilized porous hollow-fiber membrane with a density of immobilized collagenase of 52 mg/g at various residence times of the gelatin solution ranging from 0.13 to 20 min. Fourteen percent in weight of 10 g/L gelatin solution was hydrolyzed into tripeptide at a residence time of 20 min. PMID- 12892504 TI - Validation of the Hsp150 polypeptide carrier and HSP150 promoter in expression of rat alpha2,3-sialyltransferase in yeasts. AB - Heterologous glycoproteins usually do not fold properly in yeast cells and fail to leave the endoplasmic reticulum. Here we show that the Hsp150Delta polypeptide carrier promoted proper folding and secretion of the catalytic ectodomain of rat alpha2,3-sialyltransferase (ST3Ne) in Pichia pastoris. The efficiency of the Hsp150Delta carrier in P. pastoris and Saccharomyces cerevisiae was at least as high as that of the MFalpha carrier. Most of Hsp150Delta-ST3Ne and MFalpha-ST3Ne remained noncovalently attached to the cell wall via the ST3Ne portion. The strength of the HSP150 promoter was found to be comparable to that of the GAL1 promoter. PMID- 12892505 TI - Bioremediation of textile azo dyes by an aerobic bacterial consortium using a rotating biological contactor. AB - The degradation of an azo dye mixture by an aerobic bacterial consortium was studied in a rotating biological reactor. Laterite pebbles of particle size 850 microm to 1.44 mm were fixed on gramophone records using an epoxy resin on which the developed consortium was immobilized. Rate of degradation, BOD, biomass determination, enzymes involved, and fish bioassay were studied. The RBC has a high efficiency for dye degradation even at high dye concentrations (100 microg/mL) and high flow rate (36 L/h) at alkaline pH and salinity conditions normally encountered in the textile effluents. Bioassays (LD-50) using Thilapia fish in treated effluent showed that the percentage mortality was zero over a period of 96 h, whereas the mortality was 100% in untreated dye water within 26 h. Fish bioassay confirms that the effluent from RBC can be discharged safely to the environment. PMID- 12892506 TI - Monitoring of recombinant protein production using bioluminescence in a semiautomated fermentation process. AB - On-line optimization of fermentation processes can be greatly aided by the availability of information on the physiological state of the cell. The goal of our "BioLux" research project was to design a recombinant cell capable of intracellular monitoring of product synthesis and to use it as part of an automated fermentation system. A recombinant plasmid was constructed containing an inducible promoter that controls the gene coding for a model protein and the genes necessary for bioluminescence. The cells were cultured in microfermenters equipped with an on-line turbidity sensor and a specially designed on-line light sensor capable of continuous measurement of bioluminescence. Initial studies were done under simple culture conditions, and a linear correlation between luminescence and protein production was obtained. Such specially designed recombinant bioluminescent cells can potentially be applied for model-based inference of intracellular product formation, as well as for optimization and control of recombinant fermentation processes. PMID- 12892507 TI - Effect of low culture temperature on specific productivity and transcription level of anti-4-1BB antibody in recombinant Chinese hamster ovary cells. AB - Lowering the culture temperature has been suggested as a useful tool for improving the production of recombinant proteins in Chinese hamster ovary (CHO) cells. In an effort to improve anti-4-1BB antibody production in recombinant CHO (rCHO) cells, rCHO cells producing anti-4-1BB antibody (LGA31-56) were cultivated at three different temperatures, 30, 33, and 37 degrees C. Lowering the culture temperature led to suppressed cell growth, cell cycle arrest in G(0)/G(1) phase, and improved cell viability for a longer period. However, antibody production and q(Ab) were not increased at low culture temperature. The maximum antibody concentration and q(Ab) at 37 degrees C were 110.6 +/- 2.6 microg mL(-)(1) and 0.43 +/- 0.03 microg (10(6) cells h)(-)(1), respectively, whereas those at 30 degrees C were 28.3 +/- 3.8 microg mL(-)(1) and 0.44 +/- 0.07 (10(6) cells h)( )(1), respectively. Northern blot analysis revealed that lowering the culture temperature did not increase the transcription level of heavy and light chains. These results were quite in contrast with the improved production of erythropoietin, which is expressed in the same CHO host and driven by the same CMV promoters, by lowering the temperature. Taken together, the results obtained imply that the beneficial effect of low culture temperature on recombinant protein production in rCHO cells is cell-line-specific. PMID- 12892508 TI - Unstructured model for L-lysine fermentation under controlled dissolved oxygen. AB - An unstructured model was developed for batch cultivation of Corynebacterium lactofermentum (ATCC 21799) under controlled dissolved oxygen. The model is capable of predicting batch experiments performed at various initial substrate concentrations. By extending the batch culture model to a fed-batch model and using a heuristic approach to optimize the fed-batch cultivation, it is shown that fed-batch cultivation is superior to batch operation due to increased productivity at high substrate concentrations. PMID- 12892510 TI - Photodynamic therapy for the treatment of skin disease. AB - The efficacy of photodynamic therapy for the treatment of a large number of skin diseases, in particular non-malignant skin cancer, has been reported by a number of studies. It has been demonstrated to be particularly beneficial for large or multiple lesions, especially where good cosmesis is required. However for PDT to become established in routine clinical practice further comparative multi-center studies with currently accepted standard techniques are required. PMID- 12892511 TI - Optical detection and eradication of dysplastic Barrett's esophagus. AB - Dysplastic Barrett's esophagus is a condition that offers multiple diagnostic and therapeutic challenges. The diagnosis of dysplasia within Barrett's esophagus currently relies on periodic endoscopic surveillance with multiple biopsies, a methodology limited by random sampling error, inconsistent histopathologic interpretation and delay in diagnosis. Optical spectroscopic and imaging techniques have the potential to identify dysplastic or early neoplastic lesions in real-time. These diagnostic modalities are needed to enhance the endoscopic surveillance of Barrett's esophagus in the future as well as help to define lesions for endoscopic therapy. Esophagectomy has been the standard of care for Barrett's esophagus with high-grade dysplasia although it is a procedure associated with significant morbidity and mortality. Minimally invasive endoscopic ablative therapies are attractive and less morbid alternatives to esophagectomy, with promising results obtained from the use of light-activated drugs (i.e., photodynamic therapy). The combination of novel optical diagnostic techniques and therapies will provide the endoscopist with much needed tools that can considerably enhance the management of patients with Barrett's esophagus. This article reviews the current status and future prospects of optical-based modalities for diagnosis and therapy of dysplastic Barrett's esophagus. PMID- 12892512 TI - New light on the brain: The role of photosensitizing agents and laser light in the management of invasive intracranial tumors. AB - Invasive intracranial tumors, particularly malignant gliomas, are very difficult to eradicate surgically and carry a dismal prognosis. The vast majority relapse locally indicating that their cure is dependent on radical and complete local excision. However, their ability to invade and hide among normal brain tissue, our inability to visualize and detect them, the low tolerance of brain tissue to ionizing radiation and the presence of the blood brain barrier are the main causes of our failure to eradicate them. Photodynamic detection with 100% specificity and more than 80% sensitivity offers an excellent chance of visualizing camouflaged tumor nests. Also, photodynamic therapy offers a very good chance of targeted destruction of the remaining tumor cells safely following surgical excision and may double the survival of patients harboring these awful tumors. More work needs to be done to refine this promising technology to exploit it to its full potential. PMID- 12892513 TI - Clinical outcomes of photodynamic therapy for head-and-neck cancer. AB - Head-and-neck cancers not only carry poor prognoses, but also reduced quality of life for the patients. Disease control is often achieved at the expense of substantial functional loss and disfigurement. Photodynamic therapy (PDT) is particularly well suited to the treatment of head-and-neck-tumors because it has little effect on underlying functional structures and has an excellent cosmetic outcome. Studies in the past decades have shown that PDT is of similar efficacy as traditional measures in the treatment of early-stage head-and-neck cancers with an overall response rate of 85%-100% with up to 75% of the complete responses sustained at 2 years after PDT. For advanced head-and-neck cancers, studies were also conducted to evaluate the palliative effects of PDT. Overall, 58%-70% palliative benefit can be observed in these patients. Using interstitial PDT, median survival of the patients with recurrent unresectable head-and-neck cancers can be improved to 14 months (cf. 226 days by using surface illumination PDT). PDT is thus a therapeutic option that may prove a useful addition to the armamentarium of the integrated head and neck oncology team. PMID- 12892514 TI - Photodynamic therapy (PDT) in esophageal cancer: a surgical view of its indications based on 14 years experience. AB - The aim of this paper is to present the updated experience of the Yorkshire Laser Centre in PDT for esophageal cancer and to identify its role in specific subsets of patients. Also, in the light of this experience, to compare and contrast the results of PDT with appropriate subsets of patients treated in my esophageal surgery practice. 102 consecutive patients; 84 with advanced (Group A) and 18 with early (Group E) stage esophageal cancer undergoing endoscopic PDT were entered into a prospective study. Every patient had standard work up including clinical staging. PDT protocol was intravenous administration of Photofrin 2mg/kg body weight followed 24-72 hours later by endoscopic illumination using 630 nm laser light. Assessment of results was made on the basis of mortality, morbidity, patient satisfaction to treatment, symptom relief and survival. For comparison of PDT role with non PDT treated patients, reference is made to 3 previous publications comprising over 1100 patients [Moghissi, K., Br. J. Surg. 79, 935 937 (1992) (ref. 1); Sawant, D., Moghissi, K. Eur. J. Cardio-Thorac. Surg. 8, 113 117 (1994) (ref. 2); Sharpe, D. A. C., Moghissi, K. Eur. J. Cardiothorac. Surg. 10, 359-364 (1996) (ref. 3)]. There was no mortality associated with PDT. All patients expressed satisfaction to treatment. Post PDT complications consisted of photosensitivity skin reaction (sunburn) in 5 patients (5%) and esophageal stricture in 8 (8%) patients. Group A: There was significant symptom and dysphagia grade improvement. Mean survival was 9.5 months. Group E: There were no significant symptoms pre or post PDT and mean survival was 60.5 months. Comparison of PDT results in Group A with results of other palliative treatment methods, indicates that palliation can be achieved in all intraluminal cancer using PDT which is at least as good as other treatments. There is, in addition, advantage over other methods in patients with cervical esophageal cancer and in cases with re-growth of tumor obstructing previously placed stents. In early cases PDT appears capable of replicating surgical results in selected cases. PDT is an effective and safe treatment method in esophageal cancer. In advance disease it improves swallowing. In early stage disease it offers long survival and the prospect of cure in some patients. At present the role of PDT in early stage cancer should be limited to patients who are unsuitable for surgical resection. Therefore, PDT should be considered as a valid oncological option to be applied in selected cases. PMID- 12892515 TI - Raman spectral mapping in the assessment of axillary lymph nodes in breast cancer. AB - This paper is the first reported description of Raman Spectroscopy in the assessment of axillary lymph nodes in breast cancer patients. Raman Spectroscopy is an inelastic scattering spectroscopic technique appropriate for the assessment of unprocessed complex biological tissues. Spectra represent biochemical signatures of the tissue under scrutiny. The described method of Raman spectral mapping produces false-color spectral images of lymph node sections. These can be compared with standard histopathology slides and white light images of nodal tissue. This method has the potential to allow the detailed biochemical assessment of heterogeneous lymph node features, and to contribute towards the development of an optical diagnostic tool for use in a clinical setting. PMID- 12892516 TI - Standardizing dosimetry in esophageal PDT: an argument for use of centering devices and removal of misleading units. AB - A simple standardization of esophageal photodynamic therapy light dosimetry is proposed. Calculations of the effect on local treatment dose of using non centered diffusing fibers have been made and the methods of calculating light energy dose to the treatment area within a centering balloon are discussed. A requirement for centering devices and standard units of Joules per cm(2) of treatment area are indicated. PMID- 12892517 TI - Salvage reimplantation in patient with local recurrent prostate carcinoma after brachytherapy with three dimensional computed tomography-guided permanent pararectal implant. AB - Thirty-one [31] patients with local recurrent or residual adenocarcinoma of the prostate, with no evidence of distant metastases, were treated with second permanent implant using a stereotactic three dimensional system and posterior pararectal CT-guided method. All patients had extensive under local anesthesia or IV sedation pararectal CT-guided biopsy of the prostate and seminal vesicles. All patients except one had 3 month neoadjuvant androgen ablation prior to salvage reimplantation. Twenty [20] of these patients had local recurrence in the prostate and eleven [11] patients had seminal vesicle invasion which was not diagnosed nor treated with the initial brachytherapy. In addition, the patients had PSA failure or local palpable disease, "cold spots" with CT imaging or areas of dosage less than 80% of the prescribed dose with DVH (dose volume histogram). Initial prescribed dose was 120 Gy with (103)Pd loose seeds in 26 patients and 144 Gy with (125)I loose seeds in 5 patients. For the reimplant the dosage in the recurrent site was 100-144 Gy with (125)I seeds in strand in 24 patients (77%) and 100-120 Gy with (103)Pd loose seeds in 7 patients (23%). The preference of (125)I seeds in the second treatment was because only (125)I in strand was available at the time of the reimplant. Eleven [11] patients had second implant twelve to twenty-four months after the initial implant and 20 patients had after twenty-five to eighty-seven months and median follow-up was thirty months. A high level of biochemical control (87%) was achieved in all of these patients who are recognized as high risk due to local recurrence. Four [4] patients experienced grade 2 or 3 GI or GU complications and two [2] patients experienced grade 4 GI complications. Patients with local recurrent prostate cancer following initial brachytherapy including those with seminal vesicle invasion can be successfully treated with pararectal stereotactic CT-guided reimplantation. Assessment of seminal vesicle status is an essential part of staging for local recurrence. PMID- 12892518 TI - Assessment of the accuracy of a conventional simulation for radiotherapy of head and skull base tumors. AB - In this prospective study we investigated the absolute accuracy of the conventional simulation in head and skull base tumors. 41 isocenters in 40 consecutive patients with tumors of the head and skull base were included. In all cases a rigid stereotactic mask system was used for non-invasive fixation. The stereotactic ("calculated") coordinates of the isocenter were defined by the treatment planning computer. Each patient underwent a physical simulation using exclusively anatomical reference points to define the "preliminary" isocenter. The displacement between its coordinates and those of the stereotactic target point was recorded in X-, Y- and Z-direction with help of the targeting device, and the spatial error was calculated. Additionally, the patients were stratified by basal or calvarial tumor site to estimate the importance of the basal bone structures in the simulation accuracy. The influence of the learning effect on simulation accuracy was also determined. The results showed an accuracy of set-up at the linac within 1 mm in all three directions as calculated from orthogonal portal films. Mean shift of the isocenter coordinates obtained from physical simulation compared to the calculated stereotactic coordinates was 2.15 mm, 2.54 mm, and 2.69 mm for X-, Y-, and Z-direction, respectively. Mean spatial displacement amounted 5.06 mm, and the median was 4.50 mm. No significant difference could be noted between basal and calvarial location of the isocenter. A significant "learning effect" was observed with a decrease in spatial shift with increasing patient numbers. This effect was stronger in basal lesions, whereas calvarial lesions showed only a minor, insignificant effect. In conclusion, a physical simulation requires a safety margin of 5 mm in PTV definition in addition to other factors, e.g. organ movement. PMID- 12892519 TI - Establishment and characterization of a continuous cell line from pupal ovaries of Japanese oak silkworm Antheraea yamamai Guerin-Meneville. AB - Pupal ovaries of the wild oak silkworm Antheraea yamamai Guerin-Meneville were cultured in MGM-448 (Modified Grace Medium-448) medium containing 10% fetal bovine serum. After the primary culture was set up in 1988, a continuous cell line was obtained in 1991, designated as NISES-Anya-0611 (Anya-0611). The population doubling time was 54 hrs. and 19 min. at 96 passages and 88 hrs. and 29 min. at 387 passages. Spindle-shaped and spherical cells coexisted in the cell group. The cell line karyotype line was typical of lepidopteran cell lines, consisting of numerous small chromosomes. The cell line was distinguished from other lepidopteran cell lines by comparing malic enzyme, phosphoglucose isomerase, phosphoglucose mutase, and isocitric dehydrogenase isozyme patterns. The cell line was highly infected to the Antheraea yamamai nuclear polyhedrosis virus (Anya NPV). The luciferase gene of recombinant Bm NPV (BmNPVP6ETL) was able to express in the cell line, too, so that luciferase recombinant products were able to be detected in the cell body and in supernatant. The Anya NPV clone group was isolated on the cell seat using plaque purification. PMID- 12892520 TI - Enhancement of osteoblast proliferative capacity by growth factor-like molecules in bear serum. AB - The use of animal serum in cell culture is vital for providing the nutrient factors required to promote proliferation and function. Fetal calf serum has become the preferred choice because of its abundance, reasonable cost, and ability to sustain human cells in vitro. Although a wide variety of serum sources have been tested and used, little is known about the ability of serum obtained from the American black bear (Ursus americanus) to support human cell growth in culture. The American black bear, an animal comparable in size to humans, is unique in that it hibernates for mo at a time but does not experience extensive bone loss normally associated with extended immobility. The aim of this study was to analyze the effect of bear serum on human osteoblast cultures. We discovered that three of the eight bear serum samples induced significantly higher proliferation rates in osteoblasts than did fetal calf serum over a 24-h period. Osteoblasts incubated in bear serum displayed higher messenger ribonucleic acid levels for phenotype markers osteocalcin and type I collagen than did those incubated in fetal calf serum. The mitogenic activity of the bear serum was reduced when heated at 56 degrees C for 30 min before use in culture. The molecular weight of the mitogenic factors was found to be primarily greater than 50 kDa. The present work demonstrates the capability of serum from American black bears to support human osteoblast proliferation in vitro. PMID- 12892521 TI - Effects of a fat body extract on larval midgut cells and growth of lepidoptera. AB - Treatment with fat body extract (FBX) from pupae of the tobacco hornworm, Manduca sexta, caused mortality in larvae of two pest lepidopterans, the gypsy moth, Lymantria dispar, and the cotton leafworm, Spodoptera littoralis. In FBX-treated larvae, the feeding rate was depressed, causing reduced weight gain and then larval death. Their midgut showed formation of multicellular layers of midgut epidermis, indicating stem-cell hyperplasia. Hence, the integument of FBX-treated larvae had a double cuticle, indicating induction of premature molting. But radioimmunoassay measurements confirmed that the amount of ecdysteroids in FBX was too low to be responsible for the molt-inducing effects observed after treatment with FBX. With midgut stem cell cultures in vitro, addition of FBX to the culture medium stimulated cell proliferation and differentiation in a concentration-dependent manner. This effect was compared with those of insect molting hormones, ecdysone and 20-hydroxyecdysone; an ecdysteroid agonist, RH 2485; and a purified protein from FBX (multiplication factor). This article describes the mode of action of FBX and possible interplay between fat body factor(s) and insect hormones in the development and metamorphosis of the insect midgut. PMID- 12892522 TI - Maintenance of liver functions in rat hepatocytes cultured as spheroids in a rotating wall vessel. AB - Rat hepatocytes were cultured initially as spheroids on culture plates and then transferred into a rotating wall vessel (high-aspect ratio vessel [HARV]) for further culturing. Morphological evaluation based on electron microscopy showed that hepatocyte spheroids cultured for 30 d in the HARV had a compact structure with tight cell-cell junctions, numerous smooth and rough endoplasmic reticulum, intact mitochondria, and bile canaliculi lined with microvilli. The viability and differentiated properties of the hepatocytes cultured in the HARV were further substantiated by the presence of both phase I oxidation and phase II conjugation drug-metabolizing enzyme activities, as well as albumin synthesis. Homogenates prepared from freshly isolated hepatocytes and hepatocytes cultured in the HARV showed similar cytochrome P450 2B activities measured as pentoxyresorufin-O dealkylase and testosterone 16beta-hydroxylase. Further, intact hepatocytes cultured in the HARV were found to metabolize chlorzoxazone to 6 hydroxychlorzoxazone; dextromethorphan to dextrorphan, 3-methoxymorphinan, and 3 hydroxymorphinan; midazolam to 1-hydroxymidazolam and 4-hydroxymidazolam; and 7 hydroxycoumarin to its glucuronide and sulfate conjugates. In conclusion, we found that hepatocyte spheroids could be cultured in a HARV to retain cellular and physiological properties of the intact liver, including drug-metabolizing enzyme activities, plasma protein production, and long-term (1 mo) maintenance of viability and cellular function. PMID- 12892523 TI - Dilute culture media as an environmental or physiological simulant in cultured gill epithelia from freshwater rainbow trout. AB - The electrophysiological and ion-transporting properties of cultured gill epithelia from freshwater (FW) rainbow trout were examined in the presence of dilute cell culture media as an environmental or physiological simulant. Gill epithelia were cultured on cell culture inserts under symmetrical conditions (L15 apical-L15 basolateral) for 6-7 d. The following experiments were then conducted. (1) To mimic a gradual lowering of environmental salinity, apical L15 medium was progressively diluted with FW (first to 2/3 L15 for 8 h and then to 1/3 L15 for 6 h) before the introduction of apical FW (FW apical-L15 basolateral, analogous to a fish in a natural FW environment). Dilute apical media had no significant effect on the electrophysiological properties of preparations compared with symmetrical culture conditions, and no evidence for active Na(+) or Cl(-) transport was observed. Preparations subsequently exposed to apical FW exhibited a negative transepithelial potential and evidence of active Cl(-) uptake and slight Na(+) extrusion. (2) To mimic the extracellular fluid dilution that occurs in euryhaline fish after abrupt transfer from saline to FW, the osmolality or ionic strength (or both) of basolateral media was reduced by 20-40% (using either FW or FW + mannitol) while simultaneously replacing apical media with FW. Under these conditions, Na(+) and Cl(-) influx rates were low compared with efflux rates, while the Ussing flux ratio analysis generally indicated active Cl(-) uptake and Na(+) extrusion. The Na(+)-K(+) adenosine triphosphatase activity was not affected by alterations in basolateral osmolality. Our studies indicate that cultured trout gill epithelia are tolerant of media dilution from both the apical and the basolateral direction; however, neither treatment alone appeared to increase ion influx rates or stimulate active Na(+) uptake in cultured trout gill epithelia. PMID- 12892524 TI - Characterization of a white bass (Morone chrysops) embryonic cell line with epithelial features. AB - The genus Morone is an important one for U.S. aquaculture, but there has been no available cell line from this genus. We report here a cell line (the WBE line) derived from white bass embryos that has been grown for more than 80 passages over 21 mo in Dulbecco modified Eagle medium supplemented with fetal bovine serum. The WBE line showed epithelial features with positive immunohistochemical staining for cytokeratin and intercellular junctions when observed by electron microscopy. The doubling time, transformation characteristics, response to cryopreservation, and karyotype were determined. The WBE line was also tested by polymerase chain reaction to verify the species of origin and to screen for mycoplasma infection. This epithelial cell line may prove useful for the study of host-pathogen interactions that occur at the epithelial surface in this commercially important fish species. PMID- 12892525 TI - Comparison of antiproliferative effects of experimental and established antipsoriatic drugs on human keratinocytes, using a simple 96-well-plate assay. AB - Pharmacological treatments for psoriasis are generally based on antiproliferative, anti-inflammatory, or differentiation-modifying activity, or a combination of two or more of these actions. Potentially new drugs for treatment of psoriasis, which act on proliferation, can be identified by screening large compound libraries in a cell proliferation model that allows for characterization of drug effects on in vitro growth of normal human keratinocytes. High-throughput programs based on biological testing of diverse collections of compounds can rapidly identify leads for potential drug candidates in the treatment of psoriasis. In this study, we describe nonradioactive measurement of keratinocyte proliferation in the exponential growth phase in a 96-well format, using a sensitive deoxyribonucleic acid-binding dye to analyze drugs that are pharmacologically active in growth inhibition. Release of lactate dehydrogenase was used to exclude cytotoxic effects. We examined a number of compounds in a test range of 10(-7) to 10(-5) M, including known antipsoriatic drugs, and experimental drugs that are potentially useful in the treatment of psoriasis. We found strong concentration-dependent growth inhibition by dithranol, an antipsoriatic compound that is presumed to target the epidermal compartment. Methotrexate, cyclosporin A, and all-trans retinoic acid did not significantly affect proliferation at therapeutically relevant concentrations. The p38 mitogen activated protein kinase inhibitor, SB220025, and curcumin, a natural phytochemical, inhibited keratinocyte proliferation at 10(-5) M. We conclude that this assay, in combination with the previously developed assays for psoriatic differentiation, provides a useful tool for identification of antipsoriatic drugs. PMID- 12892526 TI - An efficient, nonenzymatic method for isolation and culture of murine aortic endothelial cells and their response to inflammatory stimuli. AB - Given the utility of murine models and the physiological and pathological significance of the aortic endothelium, we developed a simplified, nonenzymatic method for isolation and culture of murine aortic endothelial cells (MAECs). Aortic explants were initially cultured on fibronectin-coated plastic. Murine aortic endothelial cells migrated from the explants and proliferated. This expansion allowed for cultures to be established from the aortas of one or three mice. Murine aortic endothelial cells were then purified from expanded cultures by fluorescence-activated cell sorting for the uptake of 1,1'-dioctadecyl 3,3,3',3'-tetramethyl-indocarbocyanine perchlorate-labeled acetylated low-density lipoprotein. The majority of the cells in expanded cultures were as positive as human umbilical vein endothelial cells labeled in the same way. The most positive half of the labeled MAEC population was placed back in culture, and the cells formed "cobblestone" monolayers at confluence. Smooth muscle alpha-actin, which was present in aortic tissue and to a lesser extent in explant cultures before sorting, was not detected in selected MAECs. Western blotting and immunostaining also demonstrated the presence of the endothelial markers, platelet endothelial cell adhesion molecule-1, factor VIII-related antigen, and Bandeiraea simplicifolia lectin 1 binding. Murine aortic endothelial cells retained expected inflammatory functions: vascular cell adhesion molecule-1 protein was induced by bacterial endotoxin, and NO production was synergistically induced by the combination of endotoxin and interferon-gamma. Our simple, efficient method will facilitate investigations of aortic endothelial cell function in vitro using murine models. PMID- 12892527 TI - Effects of growth surface on differentiation of cultures of human tracheal epithelium. AB - Optical measurements from epithelial cells grown on clear solid surfaces (e.g., coverslips, petri dishes) are often compared with other measurements (e.g., short circuit current; I(sc)) obtained from cells grown on opaque porous surfaces (inserts). However, the relative levels of differentiation of cells grown under the two conditions are usually unknown. To address this issue, we grew primary cultures of human tracheal epithelium on solid surfaces or on porous inserts and compared their total levels of protein and deoxyribonucleic acid, electrical properties in Ussing chambers, and ultrastructure. To measure ion transport across cells grown on solid supports, cells were grown on inserts placed on parafilm. Later, separation of insert from parafilm allowed the cells' I(sc) to be measured in Ussing chambers. Four different media were used. Cells grown in one medium showed very low levels of differentiation on all growth supports. In the other media, growth on inserts markedly enhanced differentiation as compared with solid supports. Baseline I(sc) of cells grown on either clear or opaque inserts was at least 30 times greater than that of cells grown on solid supports, though I(sc) with clear inserts averaged approximately 30% lower than that with opaque inserts. We conclude that though differentiation of cells may vary slightly depending on the insert used, cells on any type of insert are much better differentiated than cells grown on solid surfaces. Thus, it is both possible and desirable to make all functional measurements on cells grown on clear porous supports. PMID- 12892528 TI - Effects of media on differentiation of cultured human tracheal epithelium. AB - The purpose of the this study was to find media that supported high levels of differentiation in primary cultures of human tracheal epithelium. We tested six previously described, partially defined media and three nondefined media. Cells were grown with an air interface on porous-bottomed inserts, and differentiation was assessed from electrophysiological properties, levels of total protein and deoxyribonucleic acid, and histology. In all media, cells polarized and developed tight junctions, as assessed from transepithelial electrical resistance and were better differentiated at 14 d after plating than at 7 d. The partially defined media described previously by Gray et al. (Am. J. Respir. Cell. Mol. Biol. 14:104 112; 1996) and Matsui et al. (J. Clin. Invest. 102:1125-1131; 1998) and an undefined medium containing Ultroser G serum substitute produced the most highly differentiated epithelial cells, as revealed by a high short-circuit current (I(sc)) and a ciliated, pseudostratified appearance. In other media, cells tended to be either squamous or stratified squamous, with I(sc) levels <25% of those obtained with the three optimal media. Though no key factor in the composition of the partially defined media could be identified, two of the four media with high concentrations of retinoic acid produced good differentiation. In contrast, the two media with the lowest [Ca] (0.11 mM) produced poorly differentiated cells, as did the two partially defined media with low or no retinoic acid concentration. PMID- 12892529 TI - Evidence for the isolation, growth, and characterization of malignant cells in primary cultures of human tumors. AB - Isolation and growth of malignant cells from solid tumors have often met with disappointing results. Consequently, we have developed a cell culture methodology based on ex vivo explantation of tumor tissue, with subsequent monolayer cell outgrowth. In an attempt to assess methods for detection of malignant cells in these cultures, we analyzed and compared the results of cytopathology, growth in soft agar, and detection of telomerase activity with those of standard immunohistochemistry (IHC) techniques for the detection of cytokeratins, tumor marker p53, and proliferation marker Ki-67. The sensitivity of detection of malignant cells was 85% (22/26) for cytopathological examination, 30% (3/10) for soft agar growth, and 100% (12/12) for detection of telomerase activity. From these data, we concluded that both cytopathological examination and assessment of telomerase activity contribute to the detection of malignant cells in primary cultures of human solid tumors, whereas growth in soft agar was not a good indicator of malignant cells. Although not specific for malignant cells per se, IHC detection for epithelial cell cytokeratins showed a high degree of sensitivity (100%, 23/23), whereas the sensitivity for detection of tumor marker p53 and proliferation marker Ki-67 was 30% (7/23) and 70% (16/23), respectively. These data also provide proof that malignant tumor cells, derived from a diverse number of human solid tumors, can be isolated and grown in primary cell culture. PMID- 12892530 TI - Proliferation and differentiation of Xenopus A6 cells under hypergravity as revealed by time-lapse imaging. AB - Xenopus laevis A6 cells, which are cloned epithelial cells from the Xenopus kidney, differentiate into a dome structure when the cells reach confluence. We investigated the gravitational responses of A6 cellular motility during normal differentiation and differentiation under hypergravity conditions using centrifugation (1-100 x g). Progression to dome formation was analyzed by time lapse micrography. Dome formation and increased expression of Na(+)/K(+) adenosine triphosphatase were used as markers of differentiation. Interestingly, a high rate of cellular proliferation was observed at a low level of hypergravity (5 x g). Despite this, there was no difference in the time to dome formation between the control cells at primary cell density and those that differentiated under hyper- or hypogravity conditions. In conclusion, this experiment on amphibian cells revealed that the proliferation of A6 cells was strongly affected by gravity conditions, but the differentiation step appears to be controlled by an intra- or intercellular clock. PMID- 12892531 TI - Taste receptor T1R3 is an essential molecule for the cellular recognition of the disaccharide trehalose. AB - Recently, a sweet taste receptor family, the T1R family, that recognizes some carbohydrates including sucrose was identified. Although the T1R3 molecule is known to participate in heterodimers that are used as sweet- and umami-tasting receptors, there is no evidence that T1R3 alone recognizes similar ligands. We demonstrate for the first time that the candidate sweet taste receptor T1R3 is essential for the recognition and response to the disaccharide trehalose. Our system is a valuable tool not only for understanding the relationship between sweeteners and their receptors but also for exploring the diversities of their receptors, resulting in the design of new high-potency sweeteners. PMID- 12892532 TI - Cell differentiation and p38(MAPK) cascade are inhibited in human osteoblasts cultured in a three-dimensional clinostat. AB - A three-dimensional (3D) clinostat is a device for multidirectional G force generation. By controlled rotation of two axes, a 3D clinostat cancels the cumulative gravity vector at the center of the device and produces an environment with an average of 10(-3) G over time. We cultured a human osteoblast cell line in a 3D clinostat and examined the growth properties and differentiation of the cells, including morphology, histological detection of calcification, and mitogen activated protein kinase (MAPK) cascades. In a normal 1 G condition, alkaline phosphatase (AlPase) activity was detected on day 7 of culture, bone nodules were formed on day 12, and calcium deposits were seen on day 20. In the 3D clinostat, the cells looked larger and bulged. AlPase activity was detected on day 10 of culture. However, neither bone nodules nor calcification was found in the 3D clinostat up to day 21. The expression levels of core-binding factor A1 (a transcription factor for bone formation) and osteocalcin (a bone matrix protein) increased in the control culture but decreased in culture in 3D clinostat. Phosphorylation of p38(MAPK) (p38) was repressed in culture in 3D clinostat, whereas total p38 as well as total and phosphorylated forms of extracellular signal-regulated kinases and stress-activated protein kinase/jun N-terminal kinase were not changed in the 3D clinostat. When a p38 inhibitor, SB 203580, was added to the culture medium in a normal 1 G environment, AlPase activity and formation of bone nodules and calcium deposits were strongly inhibited. On the other hand, they were inhibited only partially by a MAPK kinase inhibitor, U 0126. On the basis of these results, it is concluded that (1) osteoblast differentiation is inhibited in culture in a 3D clinostat and (2) this inhibition is mainly due to the suppression of p38 phosphorylation. PMID- 12892535 TI - Mechanism of action of newer anticonvulsants. AB - Bipolar disorder, like epilepsy, is episodic in nature. It should not be surprising, then, that anticonvulsants such as carbamazepine and valproate have proven efficacy as mood stabilizers. The newer anticonvulsants-agents like lamotrigine, gabapentin, topiramate, oxcarbazepine, and zonisamide-may also be effective treatments for bipolar disorder. Identifying an anticonvulsant for use in bipolar disorder should take into account not only the pathophysiology of bipolar disorder but also the mechanism of action of the anticonvulsant. This article will explore the mechanisms of action of the newer anticonvulsants and their relationship to the pathophysiology of bipolar disorder in an attempt to determine which of these agents might make effective mood stabilizers. PMID- 12892533 TI - Homocysteine oxidation and apoptosis: a potential cause of cleft palate. AB - Cleft palate is the most common craniofacial anomaly. Affected individuals require extensive medical and psychosocial support. Although cleft palate has a complex and poorly understood etiology, low maternal folate is known to be a risk factor for craniofacial anomalies. Folate deficiency results in elevated homocysteine levels, which may disturb palatogenesis by several mechanisms, including oxidative stress and perturbation of matrix metabolism. We examined the effect of homocysteine-induced oxidative stress on human embryonic palatal mesenchyme (HEPM) cells and demonstrated that biologically relevant levels of homocysteine (20-100 microM) with copper (10 microM) resulted in dose-dependent apoptosis, which was prevented by addition of catalase but not superoxide dismutase. Incubation of murine palates in organ culture with homocysteine (100 micro) and CuSO(4) (10 microM) resulted in a decrease in palate fusion, which was not significant. Gelatin gel zymograms of HEPM cell-conditioned media and extracts of cultured murine palates, however, showed no change in the expression or activation of pro-matrix metalloproteinase-2 with homocysteine (20 microM-1 mM) with or without CuSO(4) (10 microM). We have demonstrated that biologically relevant levels of homocysteine in combination with copper can result in apoptosis as a result of oxidative stress; therefore, homocysteine has the potential to disrupt normal palate development. PMID- 12892536 TI - Efficacy of newer anticonvulsant medications in bipolar spectrum mood disorders. AB - BACKGROUND: More treatment options for all phases of bipolar disorder are needed. While lithium, valproate, and carbamazepine remain the standard of care for treatment of bipolar disorder, many patients do not respond adequately to these treatments. Some new antiepileptic medications such as lamotrigine, gabapentin, topiramate, oxcarbazepine, tiagabine, and zonisamide are beginning to be used to treat bipolar disorder. DATA SOURCES: Evidence for effectiveness of these novel antiepileptic drugs in treating acute mania and depression as well as in preventing the recurrence of mania and depression is reviewed. A MEDLINE search (1966-2001) was performed for clinical trials that were published in English using the keywords lamotrigine, gabapentin, topiramate, oxcarbazepine, tiagabine, and zonisamide, plus the terms bipolar disorder and mania. Evidence for effectiveness of monotherapy is presented first when it is available. Data from augmentation treatment studies and open case series in which standard ratings of symptoms were employed are presented when these are the only available data. DATA SYNTHESIS: Twenty-eight reports of the efficacy of novel antiepileptic medications in bipolar disorder are reviewed. Evidence is strongest for lamotrigine monotherapy in patients with bipolar depression, in some patients with rapid-cycling bipolar disorder, and as prophylaxis. Evidence for the efficacy of topiramate in acute and refractory mania is promising but comes predominantly from open trials. Although some very small studies have found that oxcarbazepine and zonisamide may have some effectiveness for treating mania, these data are very preliminary. Results are mixed from the 2 small open trials of tiagabine. Although gabapentin is widely used in bipolar disorder, controlled data do not support the use of gabapentin as an antimanic medication or mood stabilizer. CONCLUSION: More controlled trials are needed to assess the effectiveness of novel antiepileptic medications in bipolar disorder. PMID- 12892537 TI - The efficacy of atypical antipsychotics in bipolar disorders. AB - Current treatment options for patients with bipolar spectrum disorders are expanding. New evidence in the treatment of various phases of illness of bipolar disorder is emerging. Atypical antipsychotic medications, used as monotherapy or as adjunctive therapy with mood stabilizers, have shown efficacy as well as well tolerated side effect profiles, thus becoming valuable new resources in the treatment of bipolar disorder. PMID- 12892538 TI - Weight gain in the treatment of mood disorders. AB - Overweight and obesity have become an urgent public health problem in the United States: approximately 61% of the adult population (97 million adults) are overweight or obese, where overweight is defined as a body mass index (BMI) >/= 25 and obesity is defined as a BMI >/= 30. Overweight and obesity increase the risk for developing many serious chronic diseases such as cardiovascular disease, type 2 diabetes, hypertension, dyslipidemia, and certain cancers. Increased morbidity due to obesity-related disorders begins within the normal weight range. Weight gain in adulthood per se, even in individuals who are normal weight, has deleterious health effects. Medications, particularly those commonly used in psychiatry and neurology, are a significant iatrogenic source of overweight and obesity. The weight gain potential of prescription medications should be considered in order to enhance patient compliance and reduce the risk of metabolic sequelae of weight gain. This article provides an overview of the weight-gain potential of several classes of drugs commonly used in psychiatric practice and considerations for clinicians in prescribing these medications. PMID- 12892539 TI - Newer anticonvulsants: dosing strategies and cognition in treating patients with mood disorders and epilepsy. AB - BACKGROUND: Anticonvulsants are employed to treat a variety of disorders. The most common adverse side effects of anticonvulsants are mediated via the central nervous system. Examples include sedation, dizziness, psychomotor slowing, and impairment of attention, memory, and other cognitive functions. Since multiple new anticonvulsants have been introduced in recent years, the question arises as to the frequency and magnitude of their cognitive side effects. METHODS: Experimental design flaws in assessing the cognitive effects of anticonvulsants were reviewed. A MEDLINE search of the medical literature was conducted, cross referencing terms related to cognition and anticonvulsants. Research articles were selected based on their relevance to the topic and adherence to methodological criteria. RESULTS: Incomplete information is available on the new anticonvulsants, but the present data suggest that some of the newer anticonvulsants possess favorable cognitive profiles. Also, the importance of dosing regimens and titration speed at drug initiation is discussed. CONCLUSION: All anticonvulsants possess some cognitive side effects, but differential effects can be seen. The cognitive effects of several newer anticonvulsants have been examined, but additional studies are needed to fully establish the cognitive effects of these agents. Dosage, titration rate at initiation, comedications, individual sensitivity, and underlying disease processes may influence cognitive side effects. Understanding these factors is important to maximize the benefits of anticonvulsant therapy. Cognitive side effects are one of the factors that physicians should consider in drug choice and monitoring of their patients. PMID- 12892540 TI - Decision tree for the treatment of bipolar disorder. AB - Clinicians managing patients with bipolar disorder confront a myriad of complex treatment decisions. This complexity limits the practicality of treatment guidelines, which attempt to be comprehensive. A user-friendly guide can, however, be constructed by considering only the most common early critical decision points likely to be encountered in the management of bipolar patients: new onset of an acute manic or mixed episode, interepisode treatment entry, and initial treatment for acute bipolar depression. Three general treatment principles, i.e., use proven treatments first, use a mood stabilizer in every phase of the illness, and use a multiphase treatment strategy to link current assessment with an appropriate treatment plan, can be applied to guide decision making at critical decision points that follow entry into clinical care. To guide the selection of appropriate therapeutic agents, a simple grading system can be used to evaluate the weight of evidence supporting use of various options. Multiple high-quality studies with positive results support the use of lithium, divalproex, carbamazepine, olanzapine, and haloperidol as initial intervention for acute mania; other agents with positive results in one double-blind mania trial are reasonable first-line alternatives. In the absence of high-quality evidence to guide treatment selection for nonacutely ill bipolar patients, guidelines recommend maintenance mood-stabilizer treatment. Standard antidepressant medications do not appear to add statistically significant benefit beyond that of mood stabilizers alone; lithium and lamotrigine have shown some benefit, and promising preliminary data have been presented on the antidepressant benefit of divalproex and topiramate as well. PMID- 12892549 TI - [Chronic obstructive pulmonary disease (COPD)]. PMID- 12892550 TI - [Quality of inpatient care and risk of early readmission in acute exacerbation of COPD]. AB - BACKGROUND: Chronic Obstructive Pulmonary Disease (COPD) is one of the diseases that causes more readmissions in our hospitals. The objective of our study was to establish the influence of quality of inpatient care on the risk of early readmission in the acute exacerbation of COPD. MATERIAL AND METHODS: DESIGN: case and controls study. SETTING: general acute care hospital. SUBJECTS OF STUDY: random sample of admissions with acute exacerbation of COPD and discharged alive. CASES: patients who were readmitted within 30 days with a related diagnosis. CONTROLS: patients who were not readmitted and finished alive the mentioned period. INTERVENTIONS: audit of clinical charts with evaluation of clinical severity and adherence to explicit criteria of quality of inpatient care in OCFA. Bivariate and multivariate logistic regression (LR) analysis. RESULTS: 45 cases and 45 controls were analyzed. LR model detected the risk of readmission within 1 month was explained by the higher severity of patients (Apache III: OR 1,03) and their principal diagnosis(emphysema: OR 6,9 and bronchiectasias: OR 2,2, respect to chronic bronchitis). The less score of quality of care scale was predictive of the risk of readmission in the subgroup of patients who were readmitted within 1 week (OR 0,9). CONCLUSIONS: Risk of readmission within 1 month in the acute exacerbation of COPD were explained mainly by the clinical characteristics of patients. Otherwise, readmissions within 1 week were related to quality of care, so it could be a valid indicator of quality of inpatient care. PMID- 12892551 TI - [Early hospital mortality due to infectious diseases]. AB - BACKGROUND: Infectious diseases are an important health problem. Early hospital mortality (EHM) (first 48 hours after hospital admission) give us information about the etiology and the focus of infection. This study was designed because no articles have been found about this subject. MATERIAL AND METHODS: We reviewed the medical records coded by the ICD-9-CM of all patients that suffered from EHM due to infectious diseases during the period 1992 to 1999. RESULTS: Of all the patients analyzed, 0.7% died of EHM, and of theses, 6.9% were due to an infectious disease. Median age was 73.2 years; 56.1% were men. Index of comorbidity was higher than 1 in 59,9%, and 70,7% never has been admitted to the hospital before. At admission, fever was present in 43.9%. The illness severity was 60.9% sepsis, 24.4% severe sepsis, 13.4% septic shock and 1.2% multiorgan failure. Causes of death were respiratory (76.8%; pneumonia 58.5%). Pneumonia was more frequent among aged 65 years and older (p = 0.03). In 69.5% no microbiological techniques were performed with independence of the clinical severity or the presence or absence of fever. In 85.4% the casual agent was unidentified, but in the case of isolation, gram positive was the most frequent microorganism. CONCLUSIONS: Infections are an important cause of EHM, and community-acquired respiratory tract infection (mainly pneumonia) the most frequent cause of EHM. Patients were admitted to the hospital with sepsis in 60.9%, perhaps due to a diagnostic or therapeutic delay. Among aged 65 years and older, microbiological diagnostic procedures were rarely employed. PMID- 12892552 TI - [Smoking of in-patients]. AB - OBJECTIVE: To asses the knowledge and attitude about smoking habit of in-patients in order to detect the sanitary activities that could help to decrease the prevalence of smoking. MATERIAL AND METHODS: Cross-sectional observational study of random sample (n=395) of in-patients of University Hospital Virgen Macarean, Seville, Spain. Participation rate was 89% (n=350) and the interviewer administered questionnaire was completed with epidemiological and clinical additional data. RESULTS: Sixty per cent of in-patients were smokers or former smokers. There were statistically significant differences in smoking distribution by age, gender, education, socio-economic level and admission diagnosis. Etiologic relationships between tobacco smoking and lung cancer or chronic lung disease were known by more than 90% of in-patients but other smokings health effects were not so well known: larynx (86%), oesophagus (86%) and oral cavity (81%) cancer, coronary heart disease (82%) and low birthweight (78%). Forty-five per cent of smokers began to smoke before the age of 15 and an additional 40% at 15-19 year. Forty-six per cent of smokers tried to leave this habit without success in spite of receiving medical advice of leaving smoking (61%) and the recommendations of family and friends (60%). CONCLUSIONS: Preventive campaigns against tobacco smoking must include pharmacological and psicological treatment for current smokers because an important proportion of them try to leave this habit without success. PMID- 12892553 TI - [Irritable bowel syndrome in patients under age 55 with dyspeptic syndrome]. AB - The presence of IBS was studied in 289 patients under 55 years of age with dyspeptic syndrome. The subjects were divided into two groups: group I (240 patients with DS) and group II (49 patients with DS and IBS). The mean duration of follow%shy;up was 10.8 years (130 months). IBS was diagnosed in 49 patients (17%). Females accounted for 64% and 77% of groups I and II, respectively (p < 0.05). There were no differences in mean age or the type of FDS. The patients in group II consulted more often and earlier (92% returned for consultation, and took 27 months in doing so) than those in group I (52% and 40 months)(p < 0.05). The number of diagnostic tests requested was greater in group II. Psychiatric pathology was in turn more common in group II than in group I (20 versus 7%)(p < 0.05). PMID- 12892554 TI - [Diagnosis with helical CT and echocardiography of patients with suspected chronic thromboembolic pulmonary hypertension]. AB - Chronic Thromboembolic Pulmonary Hypertension (CTEPH) is a serious disease with a very high mortality after variable periods of time. Early diagnosis of CTEPH is critical because thromboendarterectomy can improve survival, hemodimamics and functional capacity. Based on our experience we have performed a review of diagnostic techniques. Echocardiography, pulmonary angiography, fiberoptic angioscopy, helical CT an MRI. For each diagnostic procedure we have analysed its accuracy, invasiveness and easiness of use. We conclude that currently the most useful techniques are echocardiography (to asses pulmonary hypertension), and helical CT which is a good alternative to pulmonary angiography and can detect very specific findings such as dilatation of central pulmonary arteries, eccentric localized thrombi and mosaic attenuation of the pulmonary parenchyma. Even more accurate findings can be expected by the use of multislice CT, which could allow to asses the degree of narrowing of the pulmonary vessels and bronchial circulation. PMID- 12892555 TI - [Indifferentiated rheumatism and/or epileptic flatulence]. AB - A clinical case study of a 50 year-old woman brought to the Rhematology Department is presented, who showed clinical dry syndrome with immune disorders (not currently defined) and developed tumors (benign at present) and analytical autoimmunity with positive antitopoisomerase type I (formerly Scl70) confirmed. The clinical case is commented and the metheorism causes are discussed, among it, a possible variation of abdominal epilepsy as ictal flatulence, secondary to a meningioma of the right brain convexity. PMID- 12892556 TI - [Opsoclonus-myoclonus syndrome in patient with ovarian cancer]. AB - The opsoclonus-myoclonus syndrome is a rare entity as a paraneoplastic disorder usually associated to neuroblastoma in children and breast cancer or oat-cell lung carcinoma in adults. The association of opsoclonus-myoclonus syndrome and ovarian carcinoma is very unusual, to our knowledge there is only two cases reported in the literature. In both of them the opsoclonus-myoclonus syndrome preceded the neoplasy, improving with its treatment. In our patient opsoclonus began after ovarian cancer diagnosis, after chemotherapy and radiotherapy, improving with corticoid and clonazepan therapy. PMID- 12892557 TI - Expression of androgen receptor and 5alpha-reductase types 1 and 2 in early gestation fetal lung: a possible correlation with branching morphogenesis. AB - The bioactive and potent androgen 5alpha-dihydrotestosterone (DHT) has been postulated to be involved in the development of branching morphogenesis in the human fetal lung, but its expression has not been examined. We therefore examined the expression of the androgen receptor (AR) and 5alpha-reductases (type 1 and type 2), which catalyse the conversion of testosterone into DHT, in the human fetal lung using immunohistochemistry and reverse transcription-PCR (RT-PCR). Immunoreactive AR was detected predominantly in the nuclei of epithelial cells of the budding component of the early gestational fetal lung. 5alpha-Reductase type 1 immunoreactivity was detected in the cytoplasm of epithelial cells, whereas immunoreactivity for 5alpha-reductase type 2 was not detected in the samples of human fetal lung examined. RT-PCR also confirmed the presence of AR and 5alpha reductase in all fetal lung and epithelial cell lines. The results of our present study suggest that DHT may play an important role in epithelial cells, which might include precursor cells, in which both AR and 5alpha-reductases are expressed during branching morphogenesis of the human fetal lung. PMID- 12892558 TI - To b'EET or not to b'EET? That is the question! AB - Since the discovery of endothelium-derived hyperpolarizing factor (EDHF), several different candidates and pathways have been proposed as mediators of endothelium dependent hyperpolarization of vascular smooth muscle. In particular, there has been considerable support for a role for the cytochrome P450 metabolites, the epoxyeicosatrienoic acids (EETs). However, more recently, this hypothesis has come under severe scrutiny. In this issue of Clinical Science, Passauer et al. demonstrate that an EET cannot be EDHF in the human forearm, and add further to the growing belief that an EET is not a candidate for a ubiquitous EDHF. PMID- 12892559 TI - Structural basis for UCN-01 (7-hydroxystaurosporine) specificity and PDK1 (3 phosphoinositide-dependent protein kinase-1) inhibition. AB - PDK1 (3-phosphoinositide-dependent protein kinase-1) is a member of the AGC (cAMP dependent, cGMP-dependent, protein kinase C) family of protein kinases, and has a key role in insulin and growth-factor signalling through phosphorylation and subsequent activation of a number of other AGC kinase family members, such as protein kinase B. The staurosporine derivative UCN-01 (7-hydroxystaurosporine) has been reported to be a potent inhibitor for PDK1, and is currently undergoing clinical trials for the treatment of cancer. Here, we report the crystal structures of staurosporine and UCN-01 in complex with the kinase domain of PDK1. We show that, although staurosporine and UCN-01 interact with the PDK1 active site in an overall similar manner, the UCN-01 7-hydroxy group, which is not present in staurosporine, generates direct and water-mediated hydrogen bonds with active-site residues. Inhibition data from UCN-01 tested against a panel of 29 different kinases show a different pattern of inhibition compared with staurosporine. We discuss how these differences in inhibition could be attributed to specific interactions with the additional 7-hydroxy group, as well as the size of the 7-hydroxy-group-binding pocket. This information could lead to opportunities for structure-based optimization of PDK1 inhibitors. PMID- 12892560 TI - Alkylation damage repair protein O6-alkylguanine-DNA alkyltransferase from the hyperthermophiles Aquifex aeolicus and Archaeoglobus fulgidus. AB - AGT (O6-alkylguanine DNA alkyltransferase) is an important DNA-repair protein that protects cells from killing and mutagenesis by alkylating agents. The AGT genes from two extremely thermophilic organisms, the bacterium Aquifex aeolicus and the archaeon Archaeoglobus fulgidus were PCR-derived and cloned into an expression vector. The nucleotide sequence of the Aq. aeolicus AGT encodes a 201 amino-acid protein with a molecular mass of 23000 Da and Ar. fulgidus AGT codes for a 147-amino-acid protein with a molecular mass of 16718 Da. The Aq. aeolicus and Ar. fulgidus AGTs were expressed at high levels in Escherichia coli fused to an N-terminal polyhistidine tag that allowed single-step isolation and purification by metal-affinity chromatography. Both AGTs formed inclusion bodies and were not soluble under native purification conditions. Therefore AGT isolation was performed under protein-denaturation conditions in the presence of 8.0 M urea. Soluble AGT was obtained by refolding the AGT in the presence of calf thymus DNA. Both AGTs were active in repairing O6-methylguanine and, at a lower rate, O4-methylthymine in DNA. They exhibited thermostability and optimum activity at high temperature. The thermostable AGTs, particularly that from Aq. aeolicus, were readily inactivated by the low-molecular-mass inhibitor O6 benzylguanine, which is currently in clinical trials to enhance cancer chemotherapy. PMID- 12892561 TI - Direct interaction of beta-dystroglycan with F-actin. AB - Dystroglycans are essential transmembrane adhesion receptors for laminin. Alpha dystroglycan is a highly glycosylated extracellular protein that interacts with laminin in the extracellular matrix and the transmembrane region of beta dystroglycan. Beta-dystroglycan, via its cytoplasmic tail, interacts with dystrophin and utrophin and also with the actin cytoskeleton. As a part of the dystrophin-glycoprotein complex of muscles, dystroglycan is also important in maintaining sarcolemmal integrity. Mutations in dystrophin that lead to Duchenne muscular dystrophy also lead to a loss of dystroglycan from the sarcolemma, and chimaeric mice lacking muscle dystroglycan exhibit a severe muscular dystrophy phenotype. Using yeast two-hybrid analysis and biochemical and cell biological studies, we show, in the present study, that the cytoplasmic tail of beta dystroglycan interacts directly with F-actin and, furthermore, that it bundles actin filaments and induces an aberrant actin phenotype when overexpressed in cells. PMID- 12892562 TI - Stable dimeric assembly of the second membrane-spanning domain of CFTR (cystic fibrosis transmembrane conductance regulator) reconstitutes a chloride-selective pore. AB - Structural information is required to define the molecular basis for chloride conduction through CFTR (cystic fibrosis transmembrane conductance regulator). Towards this goal, we expressed MSD2, the second of the two MSDs (membrane spanning domains) of CFTR, encompassing residues 857-1158 in Sf9 cells using the baculovirus system. In Sf9 plasma membranes, MSD2 migrates as expected for a dimer in non-dissociative PAGE, and confers the appearance of an anion permeation pathway suggesting that dimeric MSD2 mediates anion flux. To assess directly the function and quaternary structure of MSD2, we purified it from Sf9 cells by virtue of its polyhistidine tag and nickel affinity. Reconstitution of MSD2 into liposomes conferred a 4,4'-di-isothiocyanostilbene-2,2'-disulphonate-inhibitable, chloride-selective electrodiffusion pathway. Further, this activity is probably mediated directly by MSD2 as reaction of its single cysteine residue (Cys866) with the thiol modifying reagent, N(alpha)(3-maleimidylpropionyl)biocytin, inhibited chloride flux. Only MSD2 dimers were labelled by N(alpha)(3 maleimidylpropionyl)biocytin, supporting the idea that only dimeric MSD2 can mediate anion flux. As a further test of this hypothesis, we conducted a second purification procedure, wherein purified dimeric and monomeric MSD2 proteins were reconstituted separately. Only proteoliposomes containing stable MSD2 dimers mediated chloride electrodiffusion, providing direct evidence that dimeric MSD2 mediates chloride channel function. In summary, we have shown that the second membrane domain of CFTR can be purified and functionally reconstituted as a chloride channel, providing a tool for probing the structural basis of chloride conduction through CFTR. PMID- 12892564 TI - Correction: The effects of aging and activity on muscle blood flow. PMID- 12892563 TI - Investigating interactions of the pentraxins serum amyloid P component and C reactive protein by mass spectrometry. AB - The oligomeric state of human SAP (serum amyloid P component) in the absence and presence of known ligands has been investigated using nanoelectrospray ionization MS. At pH 8.0, in the absence of Ca2+, SAP has been shown to consist of pentameric and decameric forms. In the presence of physiological levels of Ca2+, SAP was observed to exist primarily as a pentamer, reflecting its in vivo state. dAMP was shown not only to promote decamerization, but also to lead to decamer stacking involving up to 30 monomers. A mechanism for this finding is proposed. CRP (C-reactive protein), a pentraxin closely related to SAP, exists as a pentamer in the presence or absence of Ca2+. Pentamers of CRP and SAP were shown to form mixed decamers in Ca2+-free buffer; however, in the presence of Ca2+, this interaction was not observed. Furthermore, no exchange of monomeric subunits was observed between the SAP and CRP oligomers, suggesting a remarkable stability of the individual pentameric complexes. PMID- 12892565 TI - Cost of mental and behavioural disorders in Kenya. AB - BACKGROUND: The health and economic impact of mental and behavioural disorders (MBD) is wide-ranging, long-lasting and large. Unfortunately, unlike in developed countries where studies on the economic burden of MBD exist, there is a dearth of such studies in the African Region of the World Health Organization. Yet, a great need for such information exists for use in sensitizing policy-makers in governments and civil society about the magnitude and complexity of the economic burden of MBD. The purpose of this study was to answer the following question: From the societal perspective (specifically the families and the Ministry of Health), what is the total cost of MBD patients admitted to various public hospitals in Kenya? METHODS: Drawing information from various secondary sources, this study used standard cost-of-illness methods to estimate: (a) the direct costs, i.e. those borne by the health care system and the family in directly addressing the problem of MBD; and (b) the indirect costs, i.e. loss of productivity caused by MBD, which is borne by the individual, the family or the employer. The study was based on Kenyan public hospitals, either dedicated to care of MBD patients or with a MBD ward. RESULTS: The study revealed that: (i) in the financial year 1998/99, the Kenyan economy lost approximately US$13,350,840 due to institutionalized MBD patients; (ii) the total economic cost of MBD per admission was US$2,351; (iii) the unit cost of operating and organizing psychiatric services per admission was US$1,848; (iv) the out-of-pocket expenses borne by patients and their families per admission was US$51; and (v) the productivity loss per admission was US$453. CONCLUSIONS: There is an urgent need for research in all African countries to determine: national-level epidemiological burden of MBD, measured in terms of the prevalence, incidence, mortality, and, probably, the disability-adjusted life-years lost; and the economic burden of MBD, broken down by different productive and social sectors and occupations of patients and relatives. PMID- 12892567 TI - American physicians and dual loyalty obligations in the "war on terror". AB - BACKGROUND: Post-September 11, 2001, the U.S. government has labeled thousands of Afghan war detainees "unlawful combatants". This label effectively deprives these detainees of the protection they would receive as "prisoners of war" under international humanitarian law. Reports have emerged that indicate that thousands of detainees being held in secret military facilities outside the United States are being subjected to questionable "stress and duress" interrogation tactics by U.S. authorities. If true, American military physicians could be inadvertently becoming complicit in detainee abuse. Moreover, the American government's openly negative views towards such detainees could result in military physicians not wanting to provide reasonable care to detainees, despite it being their ethical duty to do so. DISCUSSION: This paper assesses the physician's obligations to treat war detainees in the light of relevant instruments of international humanitarian law and medical ethics. It briefly outlines how detainee abuse flourished in apartheid South Africa when state physicians became morally detached from the interests of their detainee patients. I caution U.S physicians not to let the same mindset befall them. I urge the U.S. medical community to advocate for detainee rights in the U.S, regardless of the political culture the detainee emerged from. I offer recommendations to U.S physicians facing dual loyalty conflicts of interest in the "war on terror". SUMMARY: If U.S. physicians are faced with a conflict of interest between following national policies or international principles of humanitarian law and medical ethics, they should opt to adhere to the latter when treating war detainees. It is important for the U.S. medical community to speak out against possible detainee abuse by the U.S. government. PMID- 12892566 TI - UK Back pain Exercise And Manipulation (UK BEAM) trial--national randomised trial of physical treatments for back pain in primary care: objectives, design and interventions [ISRCTN32683578]. AB - BACKGROUND: Low back pain has major health and social implications. Although there have been many randomised controlled trials of manipulation and exercise for the management of low back pain, the role of these two treatments in its routine management remains unclear. A previous trial comparing private chiropractic treatment with National Health Service (NHS) outpatient treatment, which found a benefit from chiropractic treatment, has been criticised because it did not take treatment location into account. There are data to suggest that general exercise programmes may have beneficial effects on low back pain. The UK Medical Research Council (MRC) has funded this major trial of physical treatments for back pain, based in primary care. It aims to establish if, when added to best care in general practice, a defined package of spinal manipulation and a defined programme of exercise classes (Back to Fitness) improve participant-assessed outcomes. Additionally the trial compares outcomes between participants receiving the spinal manipulation in NHS premises and in private premises. DESIGN: Randomised controlled trial using a 3 x 2 factorial design. METHODS: We sought to randomise 1350 participants with simple low back pain of at least one month's duration. These came from 14 locations across the UK, each with a cluster of 10 15 general practices that were members of the MRC General Practice Research Framework (GPRF). All practices were trained in the active management of low back pain. Participants were randomised to this form of general practice care only, or this general practice care plus manipulation, or this general practice care plus exercise, or this general practice care plus manipulation followed by exercise. Those randomised to manipulation were further randomised to receive treatment in either NHS or private premises. Follow up was by postal questionnaire one, three and 12 months after randomisation. The primary analysis will consider the main treatment effects before interactions between the two treatment packages. Economic analysis will estimate the cost per unit of health utility gained by adding either or both of the treatment packages to general practice care. PMID- 12892568 TI - Quantifying errors without random sampling. AB - BACKGROUND: All quantifications of mortality, morbidity, and other health measures involve numerous sources of error. The routine quantification of random sampling error makes it easy to forget that other sources of error can and should be quantified. When a quantification does not involve sampling, error is almost never quantified and results are often reported in ways that dramatically overstate their precision. DISCUSSION: We argue that the precision implicit in typical reporting is problematic and sketch methods for quantifying the various sources of error, building up from simple examples that can be solved analytically to more complex cases. There are straightforward ways to partially quantify the uncertainty surrounding a parameter that is not characterized by random sampling, such as limiting reported significant figures. We present simple methods for doing such quantifications, and for incorporating them into calculations. More complicated methods become necessary when multiple sources of uncertainty must be combined. We demonstrate that Monte Carlo simulation, using available software, can estimate the uncertainty resulting from complicated calculations with many sources of uncertainty. We apply the method to the current estimate of the annual incidence of foodborne illness in the United States. SUMMARY: Quantifying uncertainty from systematic errors is practical. Reporting this uncertainty would more honestly represent study results, help show the probability that estimated values fall within some critical range, and facilitate better targeting of further research. PMID- 12892569 TI - Does anybody read "evidence-based" articles? AB - BACKGROUND: The electronic version of the British Medical Journal (eBMJ) has a unique feature in that it provides an electronic record of the number of times an article has been viewed ("hits") in the week after its publication. We sought to compare the relative popularity of primary research and "evidence-based" papers against that of narrative reviews and editorials. We surveyed four broad groupings of articles in 2001: Editorials, Clinical Reviews (which are narrative reviews), Education and Debate, and Papers (which are original research articles and systematic reviews). Clinical Reviews were the most frequently viewed articles, with an average of 4148 hits per article, while Papers were less popular (average of 1168 hits per article). Systematic reviews (23 articles, average of 1190 hits per article) were visited far less often than narrative reviews. Editorials (average of 2537 hits per article) were viewed much more frequently than Papers, even where the editorial was written as an accompanying piece with a direct link to the paper. DISCUSSION: Narrative reviews and editorials are accessed more frequently than primary research papers or systematic reviews in the first week after their publication. These findings may disappoint those who believe that it is important for readers to critically appraise the primary research data. Although the technical quality of journal articles may have been helped by recommendations on structured reporting, the readability of such articles has received little attention. Authors and journal editors must take steps to make research articles and systematic reviews more attractive to readers. This may involve using simpler language, as well as innovative use of web resources to produce shorter, snappier papers, with the methodological or technical details made available elsewhere. CONCLUSION: Primary research and "evidence-based" papers seem to be less attractive to readers than narrative reviews and editorials in the first week after publication. Authors and editors should try to improve the early appeal of primary research papers. PMID- 12892570 TI - Long-term potentiation in bone--a role for glutamate in strain-induced cellular memory? AB - BACKGROUND: The adaptive response of bone cells to mechanical strain is a primary determinant of skeletal architecture and bone mass. In vivo mechanical loading induces new bone formation and increases bone mineral density whereas disuse, immobilisation and weightlessness induce bone loss. The potency of mechanical strain is such that a single brief period of loading at physiological strain magnitude is able to induce a long-lasting osteogenic response that lasts for days. Although the process of mechanotransduction in bone is incompletely understood, observations that responses to mechanical strain outlast the duration of stimulation necessitate the existence of a form of cellular memory through which transient strain episodes are recorded, interpreted and remembered by bone cells. Recent evidence supports the existence of a complex multicellular glutamate-signalling network in bone that shares functional similarities to glutamatergic neurotransmission in the central nervous system. In neurones, these signalling molecules coordinate synaptic communication required to support learning and memory formation, through a complex process of long-term potentiation. PRESENTATION OF THE HYPOTHESIS: We hypothesise that osteoblasts use a cellular mechanism similar or identical to neuronal long-term potentiation in the central nervous system to mediate long-lasting changes in osteogenesis following brief periods of mechanical strain. TESTING THE HYPOTHESIS: N-methyl-D aspartate (NMDA) receptor antagonism should inhibit the saturating response of mechanical strain and reduce the enhanced osteogenicity of segregated loading to that of an equivalent period of uninterrupted loading. Changes in alpha-amino-3 hydroxy-5-methyl-isoxazole propionate (AMPA) receptor expression, localisation and electrophysiological responses should be induced by mechanical strain and inhibited by modulators of neuronal long-term potentiation. IMPLICATIONS OF THE HYPOTHESIS: If true, this hypothesis would provide a mechanism through which the skeleton could be pharmacologically primed to enhance or retrieve the normal osteogenic response to exercise. This would form a basis through which novel therapies could be developed to target osteoporosis and other prevalent bone disorders associated with low bone mass. PMID- 12892573 TI - Small renal tumors: natural history, observation strategies and emerging modalities of energy based tumor ablation. AB - With further advances in the technology of non-invasive imaging modalities as well as their utilization, diagnosis of incidental renal tumors has increased considerably. A large proportion of these renal tumors have been small (<4 cm) masses, for which nephron sparing surgery has been proven to be effective for. The trend toward minimally invasive options in the management of renal tumors has prompted interest in energy-based ablation techniques as a possible alternative to radical or partial nephrectomy in select patients. This article will review the natural history of small renal neoplasms and the emerging modalities of energy based energy ablation such as cryoablation, radiofrequency ablation, interstitial photon radiation, interstitial laser technology, microwave ablation, and Cyberknife extracorporeal renal tissue ablation. PMID- 12892571 TI - Comparison of mode estimation methods and application in molecular clock analysis. AB - BACKGROUND: Distributions of time estimates in molecular clock studies are sometimes skewed or contain outliers. In those cases, the mode is a better estimator of the overall time of divergence than the mean or median. However, different methods are available for estimating the mode. We compared these methods in simulations to determine their strengths and weaknesses and further assessed their performance when applied to real data sets from a molecular clock study. RESULTS: We found that the half-range mode and robust parametric mode methods have a lower bias than other mode methods under a diversity of conditions. However, the half-range mode suffers from a relatively high variance and the robust parametric mode is more susceptible to bias by outliers. We determined that bootstrapping reduces the variance of both mode estimators. Application of the different methods to real data sets yielded results that were concordant with the simulations. CONCLUSION: Because the half-range mode is a simple and fast method, and produced less bias overall in our simulations, we recommend the bootstrapped version of it as a general-purpose mode estimator and suggest a bootstrap method for obtaining the standard error and 95% confidence interval of the mode. PMID- 12892574 TI - Trends in the incidence of bladder cancer in Nova Scotia: a twenty-year perspective. AB - INTRODUCTION: Bladder cancer is the most common malignant tumor of the urinary system. Tobacco smoking has been implicated as a major risk factor for the development of bladder cancer and Nova Scotia has some of the highest smoking rates in Canada. We examined trends in the incidence of bladder cancer in Nova Scotia between 1980 and 1999. MATERIALS AND METHODS: Data on incident cases of bladder cancer diagnosed in Nova Scotia over a twenty-year period (1980 - 1999) were obtained from the Nova Scotia Cancer Registry. The age- standardized incidence and mortality due to bladder cancer was calculated for both genders. Trends in the incidence of bladder cancer during the study period were analyzed for three different age groups in each gender as an estimate of birth cohort. The average annual percent change (AAPC) in incidence of bladder cancer was calculated. RESULTS: Between 1980 and 1999, 3569 cases of bladder cancer were reported (male: female = 2.9:1). The overall incidence of bladder cancer increased in both males (27.5 to 39.5 cases per 100 000) and females (7.0 to 10.7 cases per 100 000). Mortality rates were stable. There was a trend towards an increase in bladder cancer rates for all age groups analyzed, with a substantial rise occurring in females less than 65 years of age. The AAPC in incidence of bladder cancer was +1.5 for males and +2.6 for females. CONCLUSIONS: We hypothesize that the rising incidence of bladder cancer in Nova Scotia, particularly in individuals less than 65 years of age, is related to changes in cigarette smoking practices during the past century. As the population ages, we are likely to see an increased incidence of bladder cancer in females. PMID- 12892575 TI - Implementation of a PDA based program to quantify urology resident in-training experience. AB - INTRODUCTION: There currently is no simple and reliable mechanism for Residency program directors to assess how well their trainees are being exposed to all spheres of their specialty. We report on the use of hand-held personal digital assistants (PDA's) to document all clinical and academic activities of urology residents at one academic institution. MATERIALS AND METHODS: Software was developed to create customized pick lists allowing residents to record all activities on their individual PDA's. Categories included Adult Ambulatory, Pediatric Ambulatory, Adult operative, Pediatric operative, and Academic. Activities were subcategorized into detailed pick lists and time-tracking fields. Residents synchronized with a central database on a standalone hotsync server. RESULTS: In the first 8 months, 21 178 resident-hours and 5333 activities were recorded. Preliminary observations can be made regarding how residents spend the majority of their time: 28% operative, 20% self-study, 19% ward work, 10% Academics, 6% ER consultations, 5% clinic, and 4% inpatient consultations. The most common adult diagnoses encountered while attending to clinic, ward, or ER consultations were lower urinary tract symptoms, urolithiasis and hematuria. Similarly for Pediatrics: neurogenic bladder, antenatal hydronephrosis, infection, and hypospadias were most often reported. Residents reported 5,333 activities, relating to the following spheres of Urology: academics (23%), endourology (18%), oncology (15%), lower urinary tract symptoms (10%), congenital anomalies (5%), urolithiasis (5%), reconstruction (5%), and infection (3%). CONCLUSIONS: This tool provides an objective assessment of resident experience as it relates to selection of rotations, and for addressing curriculum weaknesses. It is applicable at a national level for the study of regional differences in training experience, and trends in graduate Urological education. With minimal effort it could be modified for application to other specialty training programs. PMID- 12892576 TI - Delay in surgical therapy for clinically localized prostate cancer and biochemical recurrence after radical prostatectomy. AB - BACKGROUND: In Canada, waiting times for cancer care have been increasing, particularly for patients with genitourinary malignancies. We examined whether delay from diagnosis for patients undergoing surgery for clinically localized prostate cancer affects cancer cure rates. METHODS: We conducted a historical cohort study among 645 patients who underwent radical prostatectomy between 1987 and 1997, using biochemical recurrence (PSA elevation) and metastasis as endpoints. We examined whether patients who underwent surgery >/= months (delayed surgery group) from the date of diagnosis had reduced recurrence-free survival, compared to patients who had surgery <3 months (early surgery group) from the date of diagnosis, adjusting for grade, stage and PSA level at diagnosis. RESULTS: The crude 10-year recurrence-free and metastasis-free survival rates for all patients were 71.1% (95% C.I.: 64.9% - 77.3%) and 95.3% (95% C.I.: 91.3% - 99.3%), respectively. Of the 645 patients, 189 (29.3%) had surgery >/= months after diagnosis. The median time from the date of diagnosis to surgery was 68 days (range 15 to 951 days). The 10-year recurrence-free survival was higher for patients who underwent early surgery (74.6%, 95% C.I.: 67.9% - 81.4%) compared to patients in the delayed surgery group (61.3%, 95% C.I.: 46.7% - 76.0%, p=0.05). The crude and adjusted hazard ratios for developing biochemical recurrence for patients in the delayed surgery group were 1.58 (95% C.I.: 1.0 - 2.4, p=0.04) and 1.46 (95% C.I.: 0.9 - 2.3, p=0.09), respectively, compared to patients who underwent early surgery. CONCLUSIONS: There may exist a possible relationship between delays from diagnosis for radical prostatectomy and prostate cancer cure rates. These findings may have many biases that could not be properly accounted in this retrospective analysis and larger cohort analyses will be required to confirm these findings. PMID- 12892577 TI - Malignancy in horseshoe kidneys: review and discussion of surgical implications. AB - OBJECTIVE: Horseshoe kidney is one of the most common congenital anomalies of renal structure. Not infrequently, surgical management of both benign and malignant disorders is required in patients with horseshoe kidney due to the susceptibility to certain conditions. The literature suggests a greater proclivity to certain renal tumors with this anomaly. We present three cases of malignancy in horseshoe kidneys. The unique technical challenges presented by these cases and the surgical approaches are discussed. METHODS: Three patients with tumors involving horseshoe kidneys are reviewed and their management discussed. RESULTS: Two patients were found to have renal cell carcinoma (RCC) and one had transitional carcinoma (TCC). Computed tomography (CT) and angiography were used in the work-up and preoperative planning of these cases. One patient with RCC received pre-operative renal artery embolization. Partial nephrectomy was performed in each patient with an aim at early vascular control of the tumors, identification of the collecting systems and ureters, as well as ensuring a 1 cm surgical margin. No patient required dialysis post-operatively. One patient died in the early post-operative period of a myocardial infarction; one patient developed brain metastases 18 months post-operatively, received palliative radiation and is alive 42 months after surgery; the other patient was free of disease for approximately 36 months but recently developed osseous metastases to her pelvis. CONCLUSIONS: Techniques developed for partial nephrectomy may be used in the treatment of tumors in horseshoe kidneys. Survival is related to the grade and stage of disease. PMID- 12892578 TI - Gone in 24 hours: the feasibility of performing pubovaginal sling surgery with an overnight hospital stay. AB - INTRODUCTION: Numerous management strategies are available for stress urinary incontinence, a significant problem for many women. The pubovaginal sling (PVS) with rectus fascia has proven efficacy with minimal morbidity. Historically, patients were admitted for an average of 4 days at the Ottawa Hospital. Since November 2001, patients have spent one night in hospital thereby cutting their total length of stay to less than 24 hours. This study assesses the feasibility of this approach. MATERIALS AND METHODS: Ethics approval was obtained. All patient charts were reviewed retrospectively. Exclusion criteria included women booked for combined procedures or patient refusal. Each case was reviewed to determine if admission to hospital following surgery was required or if subsequent visits to emergency occurred. The Financial Department of the Ottawa Hospital provided financial information to estimate cost savings. Each patient was contacted and administered a standard satisfaction questionnaire. RESULTS: The length of stay was less than 24 hours in all but one case. One patient required a 48-hour stay secondary to post operative nausea. No patient returned to the ER for assessment following discharge. The estimated cost savings were $2709 per case. Patients tolerated having this procedure performed on a daycare basis. CONCLUSIONS: PVS may be performed safely as short stay surgery with a high degree of patient satisfaction. This results in increased efficiency and improved utilization of increasingly limited healthcare resources. PMID- 12892579 TI - Urinary ascites secondary to forniceal rupture in a child with the Prune Belly Syndrome. AB - Despite adequate bladder catheterization, a neonate with Prune Belly Syndrome developed urinary ascites secondary to forniceal rupture. Treatment consisted of bilateral cutaneous pyelostomies. Even though most children with Prune Belly Syndrome respond to lower urinary tract drainage, a cutaneous pyelostomy may be necessary when the ureters are tortuous and do not drain adequately following bladder decompression. PMID- 12892580 TI - Herpes zoster infection: a rare cause of acute urinary retention. AB - Herpes zoster (HZ) infection has been reported as a rare cause of acute urinary retention. HZ infection involving sacral, thoracolumbar, and rarely high thoracic dermatomes is believed to occasionally cause motor and sensory neuropathy of the bladder. This is specifically achieved by the interruption of the detrusor reflex causing subsequent bladder atonia. As the course and management of this entity is quite benign, HZ should remain a diagnostic consideration in the management of urinary retention. We report a case of acute urinary retention of approximately 2.5 liters associated with HZ infection and review the proposed pathogenesis and therapeutic considerations in the management of this entity. PMID- 12892581 TI - [Early diagnosis of pancreatic cancer is essential to improving the effect of treatment]. AB - OBJECTIVES: To explore the relation between tumor TNM of pancreatic cancer and treatment, and to summarize the experience in diagnosis and treatment of pancreatic cancer. METHODS: The data on 321 patients with pancreatic cancer treated at our hospital between January 1990 and December 2000 were reviewed retrospectively. Radical resection and survival for various TNM stages were analyzed. RESULTS: The rates of radical resection for stage I, stage II, stage III, stage IVA, and stage IVB were 100%, 100%, 82.6%, 12.3% and 2.1% respectively. The 1-, 3- and 5-year survivals of stage I, II were 73.3%, 40.0% and 30.0% respectively, and higher than those of other groups. CONCLUSIONS: The patients with pancreatic cancer diagnosed early can undergo radical resection. The elevation of early diagnosis rate is significant for the improvement of survival. PMID- 12892582 TI - [Clinical significance of regional lymphadenectomy in radical resection of ductal adenocarcinoma in the pancreatic head]. AB - OBJECTIVE: To prospectively evaluate the long-term effect of pancreaticoduodenectomy with regional lymphadenectomy. METHODS: One hundred and twenty-one patients with ductal adenocarcinoma in the pancreatic head treated from 1996 to 2001 were studied prospectively. The enrollment of the patients was dependent on 7 criteria. The patients were divided into two groups: regional lymphadenectomy (group A, n = 50) and routine Whipple procedure (group B, n = 71). Their pre- and postoperative conditions, clinicopathological data, survival rates were studied. RESULTS: It was comparable between the 2 groups in age, sex, preoperative risk factors, operative management, and postoperative complication. Clinicopathological results showed no difference in tumor size and plexus invasion; but the frequency of lymph node involvement and the amount of resected lymph node in group A were significantly higher than those in group B. The rate of local recurrence was significantly higher in group A than in group B. The survival rates of 1-, 3-, 5-year in group A were 70.8%, 31.4%, 20.9%, respectively, which were higher than those in group B. No direct relations were observed between nodal involvement and survival rate. CONCLUSION: Lymphadenectomy in radical pancreaticoduodenectomy could remove lymph nodes effectively and sufficiently, and reduce the rate of local recurrence so as to improve the long term survival rate. PMID- 12892583 TI - [Preferential expression of cartilage oligomeric matrix protein in degenerating acinar cells in chronic pancreatitis and in chronic pancreatitis-like lesions]. AB - OBJECTIVES: To study cartilage oligomeric matrix protein (COMP) mRNA and protein expression in normal pancreas, chronic pancreatitis (CP), and pancreatic cancer tissues. METHODS: Tissues from 15 cases of normal pancreas, 14 cases of chronic pancreatitis and 14 cases of pancreatic cancer were analyzed by Northern blot, Western blot, in situ hybridization and immunohistochemistry. RESULTS: COMP mRNA signals and immunoreactivity were strongly present in the cytoplasm of degenerating acinar cells in CP tissues as well as in CP-like lesions in pancreatic cancer tissues. In contrast, COMP expression was weak to absent in the cytoplasm of cancer cells in pancreatic cancer tissues, and in ductal cells and islet cells in normal pancreatic tissues. CONCLUSION: COMP is preferentially expressed in degenerating acinar cells in CP and in CP-like areas in pancreatic cancer, suggesting a potential role of this molecular in acinar cell dysfunction in CP. PMID- 12892584 TI - [Combined detection of K-ras mutation and CA 19-9 level in plasma of patients with pancreatic cancer]. AB - OBJECTIVE: To confirm the effect of combined detection of K-ras gene mutation and CA 19-9 level in plasma in the diagnosis of pancreatic carcinoma. METHODS: In 33 patients without pancreatic carcinoma and 15 patients with pancreatic carcinoma, the value of K-ras mutation and CA 19-9 level were examined by enriched polymerase chain reaction (PCR) restriction fragment length polymorphism (RFLP) and radioimmunoassay respectively. The data were analysed with chi(2) test, Fisher's exact test and Kruekal-wall's test. All patients were confirmed pathologically. RESULTS: Eleven of the 15 patients with pancreatic carcinoma and 2 of 10 patients with chronic pancreatitis were found K-ras mutation. CA 19-9 level in patients with pancreatic cancer was significantly higher than that in those with benign disease but with a low specificity. The sensitivity, specificity of combined detection of K-ras gene mutation and CA 19-9 level in the plasma of patients with pancreatic cancer were 66.67% and 97%. There was a significant difference between patients with pancreatic cancer and those without pancreatic cancer. Pathological examination showed atypical hyperplasia in one patient with chronic pancreatitis who had K-ras mutation and higher CA 19-9 level. CONCLUSIONS: Combined detection of K-ras gene mutation and CA 19-9 can be used for adjuvant diagnosis of pancreatic cancer. It is more specific than detection of single K-ras gene mutation or CA 19-9 level in the plasma. The method is complementary to single detection of K-ras gene mutation or CA 19-9 level because it decreases the rate of false-positive diagnosis. PMID- 12892586 TI - [Complications of operative treatment of acetabular fractures]. AB - OBJECTIVE: To improve the effect of operative management of acetabular fractures. METHODS: One hundred and seventy eight acetabular fractures were treated operatively from August 1993 to December 2000. Their functional results and complications were analyzed. RESULTS: One hundred and twelve hips were followed up for an average of 45.7 months. Heterotopic ossification was noted in 26 hips, post-operative osteoarthritis in 22 hips, avascular necrosis of the femoral head in 8 hips, and sciatic nerve injury in 7 hips after operations. No death and infection were found in this series. CONCLUSIONS: Ectopic bone formation develops at extended ilio-femoral and Kocher-Langeneck approaches. Cartilaginous injury of the femoral head is contributable to post-operative osteoarthritis, and imperfect reduction is an important factor in the genesis of osteoarthritis. PMID- 12892585 TI - [The role of nitric oxide in lung injury associated with acute necrotizing pancreatitis]. AB - OBJECTIVE: To discuss the role of nitric oxide (NO) in lung injury associated with acute necrotizing pancreatitis (ANP). METHODS: One hundred and twenty SD rats were randomized into five groups: control group, ANP group, L-arginine (L arg) pretreatment group, L-NAME pretreatment group, and mixed pretreatment group (n = 24 for each group). Rat ANP model was induced by intraductal administration of 3% sodium taurocholate. Alveolar macrophages (AMs) were obtained by bronchoalveolar lavage. The protein content of bronchoalveolar lavage fluids (BALF), the myeloperoxidase (MPO) of lung tissue and generation of tumor necrosis factor alpha (TNFalpha)and NO by alveolar macrophages were evaluated. The expression of TNFalpha mRNA and iNOS mRNA was also measured. RESULTS: Lung injury was aggravated gradually with progression of the disease. The level of MPO of lung tissue and the protein content of BALF showed a steady increase with time and peaked at the 12(th) hour (10.8 +/- 0.6 U/g for MPO and 2,011.0 +/- 105.5 micro g/ml for protein, respectively). TNFalpha and NO secreted by AMs were elevated gradually and peaked at the 6(th) hour (1,624.2 +/- 149.2 pg/ml and 88.8 +/- 6.5 micro mol/L respectively) but decreased at the 12(th) hour. The expression of TNFalpha mRNA and iNOS mRNA was similar with the change of TNFalpha and NO. The parameters of the groups of L-arg, L-NAME and the mixed pretreatment were similar to those of ANP group. The parameters compared with those of the control group showed a significant difference (P < 0.05). The parameters of groups of L-Arg and L-NAME pretreatment in comparison with those of the ANP group showed significant difference (P < 0.05). CONCLUSIONS: Over production of NO mediated by iNOS aggravates lung injury caused by acute necrotizing pancreatitis. Administration of exogenous NOS substrate would worsen lung injury, whereas administration NOS inhibitor would alleviate lung injury. PMID- 12892588 TI - [Application of intervertebral carbon fiber cage in degenerated lumbar disorders]. AB - OBJECTIVES: To evaluate the effect of intervertebral carbon fiber cages in the treatment of degenerated lumbar disorders. METHODS: One hundred and twelve patients of degenerated lumbar disease were analyzed retrospectively with regard to the effect of posterior lumbar interbody fusion (PLIF) using intervertebral carbon fiber cage. RESULTS: In the 112 patients, disc height was 7 mm (3 to 11 mm) on average before operation and 11.2 mm (8 - 14 mm) after operation. Cobb angle was 32 degrees (21 degrees to 42 degrees ) on average before operation and 18.9 degrees (13 degrees to 21 degrees ) after operation. After follow-up for 7.8 months (4 to 13 months) on average, all operative disc spaces showed solid arthrodesis through X-ray images. Disc height was 10.9 mm (8 - 14 mm) on average, and Cobb angle was 18.5 degrees (13 degrees to 21 degrees ) on average. During the follow-up, endplate absorbing, pseudo-articulation, cage displacement, subsidence, and breaking were not found. CONCLUSION: Implantation of intervertebral carbon fiber cages is an effective method for the treatment of lumbar degenerative disease. Accurate indications and skilled techniques are required to avoid complication. PMID- 12892587 TI - [Close needling for the treatment of calcifying tendinitis]. AB - OBJECTIVE: To study the effect of close needling in the treatment of calcifying tendonitis. METHODS: Seventeen patients with calcifying tendonitis who had received close needling treatment were followed-up for 9.3 months on average. An 18-gauge or 16-gauge needle was used during needling. X-ray examination was given 4, 6, 8 and every 4 weeks after needling. SST (simple shoulder test) questionnaire, ASES (American Shoulder & Elbow Surgeon) score, constant-Murley score and UCLA score were adopted for evaluation before and after treatment. RESULTS: Calcium deposition disappeared within 4 - 20 weeks mean 9.4 weeks in 15 of the 17 patients after needling for 1 - 3 times. Average ASES score before treatment was 47.7 (34 - 59), forward elevation was 90 degrees (70 degrees - 100 degrees ), external rotation was 15 degrees (0 degrees - 30 degrees ), and internal rotation was L3-4 (L1-buttock). Average constant-Murley score before treatment was 44.6 (34 - 54), UCLA score was 11.6 (8 - 15), and numbers of questions for "yes" in SST questionnaire was 3.4 (2 - 5). After needling the average ASES score was 87.1 (72 - 91), forward elevation was 143.5 degrees (120 degrees - 160 degrees ), external rotation was 40 degrees (30 degrees - 50 degrees ), internal rotation was T(8)-T(9), constant-Murley score was 87.8 (64 - 94), UCLA score was 29.5 (19 - 33), and numbers of questions for "yes" in SST questionnaire was 9.1 (6 - 12). Significant difference was found between before and after needling (ASES: P < 0.01, forward elevation, external rotation and internal rotation: P < 0.01, constant-Murley: P < 0.01, UCLA: P < 0.01, SST: P < 0.01). CONCLUSIONS: Close needling is an effective method for the treatment of calcifying tendonitis. Symptoms and disability are greatly relieved non surgically in most of patients. Care should be taken in differential diagnosis of degenerative calcification in chronic rotator cuff disease. PMID- 12892589 TI - [Bone density in osteoarthritic femoral heads: quantitative assessment by histomorphologic and histomorphometric analysis]. AB - OBJECTIVE: To determine whether bone density is related to osteoarthritis and to compare osteoarthritis with osteoporotic fracture of the femoral neck. METHODS: All 165 femoral heads removed at joint replacement surgery were divided into 4 groups according to radiographic features of hip (osteophytes, subchondral sclerosis, cysts and femoral head deformity). The individual femoral head was divided into 5 zones histologically. Sections were studied histomorphologically, and quantitation was performed using the computer-assisted system to determine the bone density of the femoral head. RESULTS: The values of bone density in weight-bearing, super lateral non weight-bearing (SL) and inferior medial non weight-bearing (IM) areas after femoral neck fracture in patients with osteoarthritis were lower than normal. The values of bone density in weight bearing area were increased in patients with osteoarthritis in all four groups but lower than normal in one group (I), and higher than normal in other 2 groups (III, IV). The values of bone density in SL and IM areas except IM area in group IV were lower than normal (P > 0.05). The values of bone density in the central area in the 4 groups were relatively consistent and normal. The density of Haversian canals was increased in the 4 groups indicating osteoporosis of the femoral head. The average age of patients with femoral neck fracture associated with OA (group I) was significantly higher than that of group II, III, IV (P < 0.05). CONCLUSIONS: Bone density is different in the femoral head and neck. Dividing the weight bearing joint into distinct zones is a method for laboratory and clinical study. Femoral neck fracture associated with OA was caused by osteoporosis which is related to the age of the patient. The data of this study can be regarded as a potential indicator of implant/host bone relations with morphological, morphometric implications. PMID- 12892590 TI - [Treatment of intractable epilepsy with hemispherectomy]. AB - OBJECTIVE: To explore the clinical effectiveness of hemispherectomy for patients with intractable epilepsy. METHODS: Thirty-six patients with intractable epilepsy were treated by modified hemispherectomy and their clinical results were followed up after operation. RESULTS: The seizures of the 36 patients were effectively controlled and 34 (94.4%) were thoroughly controlled. Meanwhile, the dysfunction of the central nervous system in 30 patients (83.3%) was aggravated, and 5 patients (13.9%) showed better improvement than before. Neuropsychological assessment revealed much improvement in the patients. CONCLUSIONS: Modified hemispherectomy is advisable for patients with wide-range lesion of the half brain associated with intractable epilepsy. PMID- 12892591 TI - [Causes of ineffectiveness of microvascular decompression for hemifacial spasm]. AB - OBJECTIVE: To study causes of ineffectiveness of microvascular decompression (MVD) in treatment of hemifacial spasm (HFS). METHODS: Reoperative MVD was performed in 23 HFS patients with previous ineffective MVD. In the patients, the main causes of ineffectiveness included misjudgment of compressing vessels (7 patients), improper insertion of decompressing grafts (9), improper selection of grafts (5) and small grafts (2). RESULTS: Symptoms of HFS disappeared immediately after the second MVD in 21 patients and delayed in 2 patients (after 2 weeks, 6 weeks). No recurrence of HFS was noted during the follow-up period of 1.0 - 6.0 years (mean 3.4 years). CONCLUSIONS: MVD is an effective microsurgical method for treating HFS. Accurate judgement of compressing vessels and proper decompression are the key to surgical effectiveness. PMID- 12892592 TI - [Treatment of glandular cystitis]. AB - OBJECTIVE: To improve the therapeutic results of patients with glandular cystitis. METHODS: One hundred and twenty-seven patients with glandular cystitis were treated according to their different etiological factors. The therapeutic methods included anti-infection, obstruction relief, bladder irrigation, transurethral resection, partial cystectomy, total cystectomy. RESULTS: The patients who were associated stones and foreign bodies (50%) were cured spontaneously with an effective rate of 94%. The effective rate in patients with low urinary tract obstruction was 84%. Patients with simple urinary tract infection (53%) were cured spontaneously after anti-infection therapy. The effective rate, recurrence rate and malignancy rate in patients without associated diseases were 71%, 46% and 21% respectively. CONCLUSIONS: Cystitis glandularis should be treated according to different etiological factors. PMID- 12892593 TI - [Lung squamous cell carcinoma-related genes cloned by using suppression subtractive hybridization]. AB - OBJECTIVE: To identify genes differentially expressed in human lung squamous cell carcinoma (LSCC). METHODS: A subtracted cDNA library of human LSCC constructed by using suppression subtracted hybridization (SSH) method was screened. Clones representing mRNAs that were truly differentially expressed in LSCC but not in its adjacent non-cancerous tissues were identified by semi-quantitative RT-PCR in 12 patients with LSCC. Partial novel genes were detected by Northern blot. RESULTS: Ten differentially expressed gene cDNA fragments of LSSC were obtained by SSH. Among them six were known genes; two sequences were identified but their functions were unknown (hypothetical protein); two were novel (GenBank accession numbers AF363068 and AY032661, respectively). The results from semiquantitative RT-PCR showed that the transcription expression level of PPP1CB, calumenin, S100A2, HSNOV1, OCIA and AY032661 was down-regulated in some LSCC cases, while the transcription of HSP90, ferritin, gp96 and AF363068 was up-regulated in others. CONCLUSIONS: Six known genes identified by SSH technique have been implicated in the pathogenesis of lung carcinogenesis, or they are involved in immunological defense mechanism in human body. Two hypothetical proteins probably also play an important role in the pathogenesis of lung cancer. The function of the two novel genes in lung carcinogenesis are under investigation. PMID- 12892594 TI - [Single-dose ceftriaxone versus multiple-dose cefuroxime for prophylaxis of surgical site infection]. AB - OBJECTIVE: To compare the effects and pharmacoeconomics of single-dose of ceftriaxone versus 3-day cefuroxime prophylaxis in patients undergoing gastric or colorectal resection. METHODS: Three hundred and five consecutive patients with gastric or colorectal cancer from 5 medical centers were randomly divided into ceftriaxone group (n = 153, receiving intravenously 1 g ceftriaxone 0.5 - 1 h prior to operation only) and cefuroxime group (n = 152, receiving 0.75 g cefuroxime preoperatively and the same dose q8h for 3 d). The patients' intra- and postoperative status, adverse responses and infectious complications were observed and documented, and pharmacoeconomic parameters were analyzed. RESULTS: The disease distribution, operative procedures and patients' conditions in the 2 groups were comparable. No adverse responses to the test antibiotics were observed. Postoperative infectious complications occurred in 7 cases in the ceftriaxone group (4.58%) and 14 cases in the cefuroxime group (9.21%), respectively (P = 0.992), among which, 12 cases were surgical site infections (incisional, intra-abdominal): 2 cases in the ceftriaxone group (1.31%), and 10 cases in the cefuroxime group (6.58%), (chi(2) = 5.607, P = 0.018). The direct cost related to prevention and treatment of surgical site infections was 283.5 RMB in the ceftriaxone group and 811.1 RMB in the cefuroxime group (Z = 14.51, P = 0.000). CONCLUSION: Both ceftriaxone and cefuroxime are safe and effective for prevention of surgical site infections. Single-dose ceftriaxone prophylaxis is sufficient for gastric and colorectal operations, with a better cost effectiveness ratio. PMID- 12892595 TI - [Effectiveness of para-aortic lymph node dissection for advanced gastric cancer]. AB - OBJECTIVE: To assess whether any correlation exists between survival and the extent of lymph-node dissection. METHODS: D(2), D(2)(+) or D(3) radical resection was performed on 158 patients with advanced gastric cancer. Among them, 73 were subjected to para-aortic lymph node dissection, (PALD group), and the other 85 were not subjected to the operation (non-PALD group). There was no significant difference in age, gender, site of tumors, type of Borrmann and histology between the two groups. The PALD group showed more advanced diseases in term of tumor invasion, lymph node metastasis and clinicopathological stage. RESULTS: Average operation time was longer in the PALD group than in the non-PALD group [(280 +/- 93) min VS. (245 +/- 91) min, (P < 0.01)]. Blood transfusion volume was (693 +/- 324) ml in the PALD group, and (460 +/- 375) ml in the non-PALD group (P < 0.01). No anastomotic leaks, pancreatic fistula, abdominal abscess and ileus were observed in both groups. No significant complications were found except for a higher incidence of diarrhea in the PALD group. Significant difference was found in survival curve, mean and median survival time between the two groups. The difference in survival was also found between patients with positive and negative lymph node metastasis in the PALD and non-PALD groups. Again, there was significant difference in survival between positive and negative No. 16 lymph node in the PALD group. CONCLUSION: Para-aortic lymph node dissection offers a significant survival benefit to curable patients with advanced gastric cancer. It is similar to limited lymphadenectomy in morbidity and mortality. PMID- 12892596 TI - [Early superselective angiography and transarterial embolization for massive bleeding after gastrectomy]. AB - OBJECTIVE: To evaluate the efficacy of early superselective angiography and embolization in the diagnosis and treatment of massive bleeding after gastrectomy. METHODS: The clinical data of 28 patients with massive bleeding after surgery from 1980 to 2001 were retrospectively analysed. All patients underwent emergency angiography and 27 of them were treated by transcatheter embolization. RESULTS: Bleeding was controlled in 26 of the 28 patients (93%), recurrent bleeding occurred in 1, an recognized bleeding in 1, and abdominal pain in 1. There was no death. CONCLUSIONS: Transarterial embolization for massive bleeding after gastrectomy is safe and effective. It is suggested that early emergency angiography should be considered in all patients with massive gastrointestinal bleeding after gastrectomy. PMID- 12892597 TI - [Trans-sacral local resection for rectal cancer]. AB - OBJECTIVES: To evaluate the effect and the indication of local resection for rectal cancer below the peritoneal reflection. METHODS: One hundred and thirty four patients with early rectal carcinoma whose masses were >or= 5 cm or 8.0 mm. However, 6 eyes were enucleated because of the severe endophthalmitis. Most of patients sought medical care following a same procedure: village and towns medical service, county hospital, city medical center. The time interval between the first medical contact and presentation to our center was 22.9 days on average. 78% (54/69) patients attended to county hospitals, only 24% (13/54) patients got correct diagnosis of fungal keratitis. Smear examination had been undertaken only in 22% (12/54) of cases, the positive rate was 50%. In 30 patients who initially contact to city medical centers, 63% (19/30) was diagnosed as fungal keratitis, 73% (22/30) was given smear examination with positive result of 73% (16/22). The positive rate of corneal scraping for fungus was 93% (64/69). 46% (35/69) patients were prescribed antifungal agents of fluoconazole. CONCLUSION: Several factors, including ignoring traumatic history and basic examination techniques, lacking of effective ophthalmic antifungal agents, cornea donor, without qualified ophthalmologists, lead to inappropriate management in fungal keratitis. PMID- 12892603 TI - [The application of digital photography with retroillumination for lens in cataract study]. AB - OBJECTIVE: To evaluate the reliability of digital photography with retroillumination for lens as a tool to assess the degree of lens opacity, and to study the morphological features of some lens diseases using the digital images. METHODS: The cataract screener (Neitz CT-S) was connected to a computer capable of acquiring digital images of lens. The digital lens retroillumination and slit lamp images were collected in 64 eyes. Three independent separate examiners took three sets of different CT-S digital photographs in four cataract subjects. The area of lens opacification in the images was measured and analyzed using threshold adjustment in PhotoShop software and a customized program in Matlab software. RESULTS: The distinct digital images of lens cortical opacity, posterior subcapsular opacity and other lens diseases were obtained with CT-S computer digital photography, which images were much better than slit lamp sectional digital images. The sensitivity and specificity of measuring lens opacification with CT-S digital images were 90% and 94% respectively. However, the digital lens retroillumination images were not as sensitive as the slit lamp images for nuclear cataracts. The variation coefficients of lens opacity area in the three sets of CT-S digital images in the same subject, which were taken by the independent examiners, were from 2.23% to 10.86%. The cortical opacification in CT-S digital images was linear, cuniform or clustered aggregate shadow configuration. The posterior subcapsular opacification was discal vacuolar or granular appearance. CONCLUSIONS: The reproducibility of the CT-S digital images taken by the independent examiners is excellent. This technique can objectively show cortical or posterior subcapsular opacities, but nuclear cataracts. PMID- 12892604 TI - [Changes of lens capsule after phacoemulsification]. AB - OBJECTIVE: To observe the changes in the lens capsule after phacoemulsification and its effects on the position of intraocular lens (IOL) and to demonstrate the best position of IOL in relation to the anterior capsule. METHODS: The present series comprised 141 eyes in 127 patients underwent phacoemulsification and foldable lens implantation with 5 mm continuous curvilinear capsulorhexis. The changes in the lens capsule and the position of IOL with dilated pupil were observed using slit lamp microscope 3 months after the operation. RESULTS: Capsular bag shrinkage and white Soemmering ring along the margin of capsulorhexis were observed in all cases. Three different kinds of relationship between anterior capsule opening and the IOL optic surface were found, including the non-capture (68 eyes), partial-capture (52 eyes) and total-capture (21 eyes) relationships. The incidence of central posterior capsule opacification (PCO) was significantly higher in the total-capture group (47.6%) than that in the non capture (11.7%) and partial-capture (21.2%) groups (P < 0.01). IOL in well centered position was significantly higher in the non-capture (91.2%) and total capture (81.0%) groups than that in the partial-capture group (42.3%) (P < 0.01). CONCLUSION: In the present study, various changes in the lens capsule were observed after phacoemulsification and foldable lens implantation. It is suggested that keeping the IOL in a non-capture position is a key point to avoid the occurrence of PCO and IOL decentration. PMID- 12892605 TI - [The recovery of binocular vision in the early post-operative stage of senile cataract patient]. AB - OBJECTIVE: To study the recovery of binocular vision and the risk factors that affect the recovery in the early post-operative stage of senile cataract patients. METHODS: Patients undergone extracapsular cataract extraction with intraocular lens implantation (ECCE group) or cataract phacoemulsification with intraocular lens implantation (PHACO group) were examined for their corrected visual acuity, refractive power, simultaneous perception, fusion, near and far distance stereoacuity. RESULTS: Before operation, the visual acuity was worse in the ECCE group than that in the PHACO group (chi(2) = 9.769, P < 0.05); After operation, the visual acuity between ECCE group and PHACO group was not statistically significant (chi(2) = 0.52 for operated eyes, P > 0.05, chi(2) = 3.52 for non-operated eyes, P > 0.05). The cylinder anisometropia in ECCE group was worse than the PHACO group (chi(2) = 12.496, P < 0.01). All patients obtained simultaneous perception and fusion sense. There was more or less far-distance stereoacuity 83.3% in ECCE group and 94.7% in PHACO group, which was no statistical significant (chi(2) = 1.456, P > 0.05). Foveal near stereoacuity was established in ECCE group (2.8%) and in PHACO group (21.1%), which were statistically significant (chi(2) = 5.029, P < 0.05). CONCLUSIONS: There is incomplete recovery of binocular vision in the early post-operative stage of senile cataract patients. The vision depression, especially monocular depression before surgery and the anisometropia after operation may affect the recovery of binocular vision. PMID- 12892606 TI - [Selection of IOL power formula for patients with cataract and high myopia]. AB - OBJECTIVE: To compare the accuracy of two IOL power formulas (SRK-T and SRK-II) in order to improve the accuracy of predicative IOL powers for patients with cataract and high myopia. METHODS: (1) Patient's axial lengths of the eye, K readings and A-constants were input to the SRK-T and SRK-II formulas. The differences in predicative refraction between the two formulas were compared. (2) Eight six patients (130 eyes) with cataract and high myopia underwent phacoemulsification and posterior chamber IOL implantation were studied. These patients were randomly divided into two groups and received IOL whose power was determined by either SRK-T or SRK-II. Three months after IOL implantation, the postoperative refractions were recorded and compared with predictive refractions. Patients were followed-up for 3 to 24 months. RESULTS: (1) When K-reading was 43.00 D, there was no statistically significant difference between the results from these two formulas (P > 0.05) and there was no correlation between the difference and the axial lengths (P > 0.05). In eyes with K-reading at 39.00 D, 41.00 D, 45.00 D, or 47.00 D, there was statistically significant difference between the results from these two formulas (P < 0.05). The difference between the two formulas was in proportional to the axial length (P < 0.05). (2) The mean error between the postoperative refraction and predictive refraction determined by SRK-T and SRK-II formula was 0.08 D and -0.79 D, respectively. The difference between these two formulas was statistically significant (P < 0.05). CONCLUSION: Selection of IOL power formula should consider the axial length, K-reading and other related factors. SRK-T formula is appropriate for high myopia eyes with axial length longer than 26.0 mm. In this group, SRK-T formula is much more accurate than the SRK-II formula. PMID- 12892607 TI - [Effect of the different diameter of posterior capsulotomy on visual field]. AB - OBJECTIVE: The purpose of this study is to determine the influence of two different diameter of posterior capsulotomy on visual field. METHODS: Forty patients with posterior capsule opacification (PCO) of grade II after phacoemulsification and intraocular lens implantation were randomly divided into two groups. 20 cases received laser capsulotomy with 2 mm diameter. Other 20 cases received laser capsulotomy with 4 mm diameter. One and half months after laser capsulotomy, vision field of every patient was examined using the automated perimetry for central 30 degrees and peripheral 60 degrees. Visual field in 21 patients with a transparent posterior capsule after cataract surgery was also evaluated by the same automated perimetry. RESULTS: There were significant reductions in the total visual field sensitivity, the mean deviation (MD) and the pattern standard deviation (PSD) in 2 mm diameter capsulotomy group compared with 4 mm diameter capsulotomy group (P < 0.001). There were no significant difference in visual field between 4 mm capsulotomy group and the transparent posterior capsule group (P > 0.05). CONCLUSIONS: Better visual acuity could be obtained after laser capsulotomy with 2 mm capsulotomy, however the generalized sensitivity reduction and irregular localized depression of visual island might occurred. 4 mm diameter capsulotomy may be a better choice in laser capsulotomy in patients with PCO. PMID- 12892608 TI - [Study on the experimental model of krypton laser-induced choroidal neovascularization in the rats]. AB - OBJECTIVE: To evaluate the feasibility of krypton laser-induced choroidal neovascularization (CNV) model in the Brown Norway rats in order to establish the foundation for studying the mechanism of CNV and for the development of treatment for CNV. METHODS: Experimental eyes in 33 rats were received a series of 10 krypton red laser (647 nm) lesions per eye (360 mW, 50 micro m, 0.05 s). Fundus fluorescein angiography (FFA) and indocyanine green angiography (ICGA) were performed, then the rats were sacrificed immediately, the eyes were enucleated and processed for histopathologic examination and transmission electron microscopy on days 3, 7, 14, 21, 28 and 56. RESULTS: CNV was firstly appeared on day 7 after photocoagulation, reaching the peak on day 21. The incidence of CNV was 76.0%. Disciform leakage staining appeared in the FFA. ICGA showed that the disciform area was filled with CNV. CNV was ascertained by light microscopy and electron microscopy. The thickness of laser-induced CNV was increased from day 7 to day 21 (P < 0.01) and no significant progress occurred after day 21 (P > 0.05). CONCLUSIONS: The present studies demonstrated that krypton laser photocoagulation could be used to produce choroidal neovascularization experimental model in the Brown Norway rat. CNV can be induced rapidly by the method, persists for a long period and at a high successful rate. PMID- 12892609 TI - [Neuroprotection of nerve growth factor in experiment retinal detachment]. AB - OBJECTIVE: To study the neuroprotection by nerve growth factor (NGF) in retina in experimental retinal detachment (RD). METHODS: Thirty one Sprague-Dawley rats were used as RD animal model by injected 0.1% sodium hyaluronate into the sub neuroretina space. Those rats were divided into NGF experimental group, placebo group, normal control groups and pathologic control group. NGF 1 g/L/eye was injected into vitreous every 4 days in NGF experimental group and 5 micro l of PBS intravitreal injection was used as control after RD. On postoperative time 1/2 d, 1 d, 2 d, 4 d, 8 d, 16 d, 32 d, rats were sacrificed and eyes were enucleated. The effect of NGF on retina was assessed by electron, light microscopy and cell counters. RESULTS: Histology studies showed markedly changes in photoreceptor cells, bipolar cells and ganglion cells after RD, included depletion and shorten of inner and outer segments, disturbance and thinness of inner and outer nuclear layer and swelling of ganglion cells in NGF treated group and control. Changes in the microscopic examination in NGF group showed significantly less than that in control groups, especially in inner and outer segments of photoreceptor cells. In reattached retina, the tissue structure and cell morphology showed more close to normal in NGF group than that in RD control groups. Cell counters showed the cell numbers decreased both in NGF groups and RD control groups. When retina reattached, the cell number in experimental groups showed significantly lower than that in normal control group (P < 0.05). Nuclear number of outernuclear layer and ganglion cell layer in NGF groups was greater than that in RD control groups (P < 0.05). CONCLUSION: Intravitreal injection exogenous NGF partly protects retinal cells from degeneration in experimental RD and help recovery of retinal cells after retina reattached. PMID- 12892610 TI - [Prevalence of obstructive sleep apnea-hypopnea syndrome in Chinese adults aged over 30 yr in Shanghai]. AB - OBJECTIVE: To investigate the prevalence of obstructive sleep apnea-hypopnea syndrome (OSAHS) in adults aged over 30 yr in Shanghai, to provide data for estimating the burden of this disease and making strategies for treatment and prevention of this disorder. METHODS: 9,600 subjects (aged >or= 30 yr) were derived from a random sample of the population living in six streets, four districts in Shanghai. They were asked to answer the questions from a questionnaire concerning snoring, daytime sleepiness, and habits such as smoking and drinking, etc. According to the degree of snoring, 150 moderate and severe snorers were selected to be tested by portable PSG for whole night data. The prevalence of OSAHS was conservatively estimated from the result. RESULTS: 8,081 subjects (84.18%) had answered the questions, among them 6,826 (84.47%) were available for evaluation. The prevalence of ever snoring was 57.11%, moderate and severe ones 23.73%. Before the age of 70, the prevalence of snoring increased with age, and higher in males than females, particularly for moderate and severe snoring. Smoking and drinking were associated with the severity of snoring. The prevalence was higher in women post-menopause and with irregular menstruation. ESS was significantly different among snorers with various severities. When AHI >or= 5 was taken as the cut-point, the prevalence of OSAHS was 20.39%. After excessive daytime sleepiness (EDS) definited as ESS >or= 9 was added as a criterian, the conservatively estimated prevalence of OSAHS was 3.62%. CONCLUSIONS: The estimated prevalence of OSAHS in adults over 30 yr old was 3.62%. This can be a great burden to public health and implies that the disease needs better understanding and study. PMID- 12892611 TI - [Epidemiologic study on sleep apnea-hypopnea syndrome by home investigation in Chengde city]. AB - OBJECTIVE: To investigate the prevalence of sleep apnea-hypopnea syndrome (SAHS) among adults aged over 30 yr in Chengde city of Hebei province. METHODS: 1 168 subjects (over 30 yr) were derived from a random sample of the population among the Shuangqiao district in Chengde city. All subjects were asked at home to answer questions about their snoring, daytime sleepiness, and habits of smoking and drinking. According to the degree of snoring, 127 moderate and severe snorers were measured by portable PSG for a whole night and the prevalence of SAHS was estimated according to AHI and ESS scores. RESULTS: 1 168 subjects (95.42%) answered the questions. The prevalence of snoring was 53.76%. The prevalence of moderate and severe snoring (>or= second degree) was 28.25%. The prevalence of snoring increased with age before the age of 70. The prevalence of snoring in males was significantly higher than that in females. Smoking and drinking groups were associated with a higher prevalence of snoring. The prevalence of snoring was higher in drivers (42.00%) than that in other populations. The estimated prevalence of SAHS according to both the AHI > 5 and the ESS >or= 9 was 4.63%. CONCLUSIONS: The estimated prevalence of SAHS among adults aged over 30 yr in Chengde city was 4.63%. It means that SAHS needs better understanding and more study. PMID- 12892612 TI - [Immediate intraoral adaptation of mandibular advancing appliances of thermoplastic material for the treatment of obstructive sleep apnea-hypopnea syndrome]. AB - OBJECTIVE: To appraise the effect of immediate intraoral adaptation of mandibular advancing appliances (MAA) consisting of thermoplastic material for the treatment of obstructive sleep apnea-hypopnea syndrome (OSAHS). METHODS: 45 patients with OSAHS was treated with MAA that was directly moulded intraorally before sleep. Polysomnography (PSG) was performed and questionnaires recorded prior to and 4 weeks after treatment with MAA. RESULTS: The efficiency of MAA for OSAHS was 82%. The medial apnea-hypopnoea index decreased from 38 +/- 22 to 11 +/- 9 events per hour (P < 0.001); the medial duration of apnea decreased from 22 +/- 8 to 15 +/- 11 second (P < 0.001); the minimum SaO(2) increased from (72 +/- 11)% to (83 +/- 9)% (P < 0.001), and the medial snore index decreased from 304 +/- 105 to 108 +/- 113 events per hour (P < 0.001). The patients' daytime sleepiness was alleviated, and the majority of the patients could acclimatize themselves to MAA in 3 to 7 days. CONCLUSIONS: The MAA consisting of thermoplastic material improved OSAHS. It is immediately adaptable and less expensive, and should be considered as a feasible medical treatment. PMID- 12892613 TI - [Retrospective analysis of the studies on sleep apnea-hypopnea syndrome in the past twenty years in our country]. AB - OBJECTIVE: To summarize and evaluate the studies on sleep apnea-hypopnea syndrome (SAHA) in our country. METHOD: Articles on the study of SAHS published in major journals of our country from January, 1982 to June, 2002 were evaluated retrospectively. RESULTS: 262 articles were recruited. More papers covering more aspects of this disorder were getting published in the past 20 years. However, basic studies, multidisciplinary studies, more effective therapies and large size, multicenter, and prospective trials were lacking. The study of SAHS in different provinces did not reach the same level. CONCLUSION: Large sampled, multicenter and multidisciplinary prospective trials are needed to study more effective and acceptable therapies as well as the basic mechanism of SAHS. PMID- 12892614 TI - [The role of the static P-V curve under zero end-expiratory pressure in sustained inflation in patients with acute respiratory distress syndrome]. AB - OBJECTIVE: To evaluate the value of static P-V curve under zero end-expiratory pressure (ZEEP) in predicting the effect of sustained inflation (SI) on hemodynamics, oxygenation and alveolar recruitment in patients with acute respiratory distress syndrome (ARDS). METHODS: Static P-V curve was measured under positive end-expiratory pressure (PEEP) in all the patients 2 h afer PEEP was applied. Patients who experienced more than 20% increase in PaO(2)/FiO(2) were considered as responders to SI. RESULTS: (1) The static P-V curves in responders consistently showed a concave pattern with c-2d >or= 0 cm H(2)O (1 cm H(2)O = 0.098 kpa) and c >or= 18 cm H(2)O, while those in non-responders showed a convex pattern with c-2d < 0 cm H(2)O or c < 18 cm H(2)O. (2) After SI, decrease of Q(s)/Q(t) (P = 0.006) was found in responders, but not in non-responders (P = 0.339). The amount of recruited volume was significantly higher in responders than in non-responders after SI [(241 +/- 111) ml vs (29 +/- 46) ml, P = 0.036]. CONCLUSION: The static P-V curves under ZEEP exhibited different patterns in responders and non-responders to SI in ARDS patients, and may be of value in predicting the response to SI. PMID- 12892615 TI - [A study on the association of 3'UTR polymorphisms of NRAMP1 gene with susceptibility to tuberculosis in Hans]. AB - OBJECTIVE: To determine whether 3'UTR polymorphisms of the NRAMP1 gene are associated with tuberculosis in Hans. METHODS: 3'UTR polymorphisms of NRAMP1 gene were typed by PCR-RFLP among 147 patients with active tuberculosis and 145 healthy individuals. The relationship between 3'UTR polymorphisms and susceptibility to tuberculosis was studied, and cases were grouped according to genotypes. RESULTS: In the tuberculosis patients, genotype TGTG/TGTG, TGTG/TGTG deleted, and TGTG deleted/TGTG deleted were observed in 95, 50 and 2 cases respectively, while the genotypes of the healthy controls were TGTG/TGTG in 115, TGTG/TGTG deleted in 29 and TGTG deleted/TGTG deleted in 1 case. The frequency of the genotype TGTG/TGTG was found more often among controls than that in patients (chi(2) = 7.79, P < 0.01). The frequency of allele TGTG and the frequency of variant allele were 0.85 and 0.15 respectively. CONCLUSIONS: 3'UTR polymorphisms of NRAMP1 gene are associated with susceptibility to tuberculosis in Hans. The variant allele observed in Hans is more common than that in Caucasians. These observations might explain in part why Hans have greater susceptibility to tuberculosis than Caucasians. PMID- 12892616 TI - [The effect of murine interferon-gamma transgene expression on bleomycin-induced pulmonary fibrosis in mice]. AB - OBJECTIVE: To investigate the effect of murine interferon-gamma (IFN-gamma) transgene expression on bleomycin-induced pulmonary fibrosis in mice. METHODS: 6 8 week aged pathogen-free male ICR mice were treated with bleomycin (3 mg/kg weight in 70 micro l) by intranasal instillation on day 0 in the model group. Adenoviral vector with murine IFN-gamma cDNA (AdCMVmIFN-gamma) 5 x 10(8) plague forming unit (pfu) was administrated via nastril instillation 48 h before bleomycin treatment in the preventive therapeutic group, but 72 h after bleomycin treatment in the therapeutic group. Mice treated with a same volume of normal saline (NS) and a same dosage of sham recombinant adenoviruses (AdCMVNull) served as controls. The animals were weighted on day 0, 7, 14, and sacrificed on day 14. Bronchial alveolar lavage fluid (BALF) and lungs were recovered. The right lung was stained with either hematoxylin-eosin or masson. The left lung was weighted, and its content of hydroxyproline (HYP) was measured by HCl acid hydrosis method. The IFN-gamma concentration in BALF was determined with enzyme-linked immunosorbent assay. RESULTS: The concentrations of IFN-gamma in BALF from the AdCMVmIFN-gamma treated groups were remarkably increased, (21,250 +/- 6,497) pg/ml and (21,000 +/- 5,451) pg/ml in the IFN-gamma transgenetic preventive therapeutic group and the therapeutic group respectively. IFN-gamma was undetectable in BALF from other groups. Mice in the two groups treated with AdCMVmIFN-gamma had statistically significant weight loss (P < 0.05), higher HYP content (P < 0.05), and tended to have more severe alveolitis and pulmonary fibrosis (P = 0.07) as compared with those in other groups. CONCLUSIONS: (1) mIFN gamma could be overexpressed in airway and alveolar epithelium by locally administrated AdCMVmIFN-gamma. (2) Early mIFN-gamma transgene expression via adenoviral vector in this bleomycin model aggravated alveolitis and fibrosis to some degree. PMID- 12892617 TI - [Electrical cardioversion for atrial fibrillation. Are there indications left after the AFFIRM study?]. PMID- 12892618 TI - [Acute coronary syndromes and multivessel coronary artery disease]. PMID- 12892619 TI - [Current status of secondary prevention and cardiac rehabilitation programs in Spain]. PMID- 12892620 TI - [Importance of severe lesions left untreated in patients with acute coronary syndromes and angioplasty of the culprit lesion]. AB - INTRODUCTION AND OBJECTIVES: Patients with acute coronary syndromes may have significantly stenotic nonculprit lesions that do not show complex lesion morphology. We investigated whether these lesions were prone to become unstable since they exist within a prothrombotic and inflammatory systemic milieu. PATIENTS AND METHOD: We evaluated the clinical course of 150 patients after successful angioplasty of a culprit lesion: 75 patients with a severely stenotic but uncomplicated nonculprit lesion (group A) and 75 patients without these lesions (group B). RESULTS: In group A, 1 patient (1.3%) required angioplasty of an initially nonculprit lesion, and in group B, 2 patients (2.6%) died in cardiogenic shock. After 1 year of follow-up, in group A, 4 patients (5.3%) died (cardiac deaths), 1 patient (1.3%) had a myocardial infarction, and 10 patients (13.3%) underwent a repeat revascularization procedure, which in 6 cases (8%) was angioplasty of an initially nonculprit lesion. In all 6 patients with angioplasty of the initially nonculprit lesion, revascularization was done within the first 4 months and was indicated for unstable angina. In group B, 1 patient (1.3%) died (noncardiac death) and 2 patients (2.6%) underwent a repeat revascularization procedure because of restenosis. Survival curves were significantly different between both groups. Belonging to group A was the only independent predictor for events, and within this group location of the lesion in the left anterior descending artery was the main predictor. CONCLUSIONS: The presence of nonculprit lesions of uncomplicated morphology at the time of a percutaneous revascularization procedure for a culprit lesion in patients with acute coronary syndrome is a short- and middle-term predictor of a moderate rate of recurrent events when these initially innocuous lesions become unstable. PMID- 12892621 TI - [Differences in clinical presentation between subjects with a phenotype of familial hypercholesterolemia determined by defects in the LDL-receptor and defects in Apo B-100]. AB - INTRODUCTION AND OBJECTIVES: Familial hypercholesterolemia and familial defective Apo B-100 are phenotypically indistinguishable. At present they can be distinguished by genetic analysis. PATIENTS AND METHODc We compared the clinical features of 13 subjects with familial defective Apo B-100 and 39 subjects with familial hypercholesterolemia. We used data from first degree relatives to compare morbidity and mortality between the two groups. RESULTS: We found statistically significant differences in total cholesterol and LDL cholesterol, which were lower in the familial defective Apo B-100 group (TC = 357 37.3 mg/dl vs 415 79.7 mg/dl and LDLc = 270 34.2 mg/dl vs 355 72.4 mg/dl). We found no differences in xanthomas, corneal arcus, smoking status, vascular events, blood pressure, BMI or waist/hip ratio. There were no differences between the two groups in the proportions of patients with cardiovascular disease or patients who died. We found statistically significant differences between the groups (p = 0.023) in the mean age at first vascular event (familial hypercholesterolemia and first degree relatives: 45.3 19.9 years; familial defective Apo B-100 and first degree relatives: 51.5 20.8 years). CONCLUSIONS: We conclude that familial defective Apo B-100 results in clinically milder hypercholesterolemia than familial hypercholesterolemia, and that discerning between them could be helpful to stratify the risk in persons with hereditary hypercholesterolemia. PMID- 12892622 TI - [Incorporation of cardiac rehabilitation programs and their characteristics in the Spanish National Health Service]. AB - OBJECTIVES: To estimate the degree of incorporation of cardiac rehabilitation in the Spanish National Health Service, to describe the characteristics of the programs, and to report on the opinions of those responsible for them regarding their progress. PATIENTS AND METHOD: Cardiac rehabilitation centers were identified from different sources. A questionnaire which included items about coverage, resources, activities and services, selection of patients, and opinions was mailed to the heads of all units. RESULTS: Twelve public centers with cardiac rehabilitation programs were identified. Cardiac rehabilitation was offered to 53% of all eligible patients. All units treated patients with myocardial infarction, 64% treated those with heart failure; and 60% high risk patients. Approximately 10-19% of all patients were women. The physicians involved most frequently in programs were cardiologists; nonmedical professionals who participated most often were physiotherapists, and 64% of all units had a staff psychologist. Phase II rehabilitation was provided by all units, and phase III treatment was provided mainly by units that operated in coordination with out patient services (45%). All units provided physical exercise training and counseling about the disease and risk factors, and 73% of them provided psychological support. The main reasons cited for providing rehabilitation were its efficacy and ability to prevent illness; and the main barriers to more widespread use were lack of resources and support. About three-fourths (73%) of all doctors interviewed thought that primary health care centers could play an important role in rehabilitation programs. CONCLUSIONS: Cardiac rehabilitation is poorly implemented in the Spanish National Health Service. The most significant differences between programs were related to the inclusion of high risk patients and with a diagnosis other than myocardial infarction, coordination with out patient services, and provision of phase III rehabilitation. PMID- 12892623 TI - [Prevalence of main cardiovascular risk factors in women from Biscay]. AB - INTRODUCTION AND OBJECTIVES: The aim of this study was to analyze the prevalence of principal cardiovascular risk factors in the female population from Biscay (northern Spain). PATIENTS AND METHOD: We selected a random representative sample of 1,317 women aged between 16 and 65 years from this province. For each participant we recorded the following parameters: weight and height, physical activity, smoking, blood pressure, glycemia, total cholesterol, triglycerides, HDL-cholesterol and LDL-cholesterol. RESULTS: A total of 1,100 women (mean age 39.83 14 years) participated. Regarding physical activity, 31.9% of the women had a sedentary lifestyle and 48.4% did not exercise during leisure time. The prevalence of smoking was 31.9%. We found a mean body mass index of 24.9 4.6 kg/m2, and 42.4% of the women were overweight. The prevalence of hypertension was 13.1%, hypertension being defined as a mean systolic blood pressure > or = 160 mmHg, diastolic blood pressure > or = 95 mmHg, current antihypertensive treatment, or any combination of these criteria. When a cutoff value of > or = 140/90 mmHg was used, the prevalence increased to 26.7%. Total cholesterol values were > or = 240 mg/dl in 26.2%, triglyceride levels were > or = 200 mg/dl in 2.6%, LDL-cholesterol was > or = 160 mg/dl in 26.8%, and HDL-cholesterol values were < 45 mg/dl in 12.2%. The prevalence of hyperglycemia was 3.3%. CONCLUSIONS: The prevalences of main cardiovascular risk factors were similar to those in other Spanish studies. Except for smoking, the rest of these risk factors increased with age. Long-term measures should be adopted to modify dietary habits and lifestyles to obtain improvements in the cardiovascular risk profile. PMID- 12892624 TI - [Effect of time elapsed from spontaneous syncope on the diagnostic performance of the head-up tilt table test]. AB - OBJECTIVES: The propensity for spontaneous and tilt-induced neurocardiogenic syncope may exhibit temporal variations. Therefore the diagnostic performance of the head-up tilt test could be improved if it were performed soon after the occurrence of spontaneous syncope. The objective of this study was to assess whether the time interval between the last syncopal episode and tilt table testing influenced the outcome of the test. PATIENTS AND METHOD: Three hundred and fifteen patients undergoing diagnostic tilt table testing potentiated with nitroglycerin for suspected neurocardiogenic syncope were included in the study. The time between the last spontaneous syncope and the tilt table test was recorded and its relationship with the results of the test was analyzed. RESULTS: The tilt table test was positive in 211 patients (67.0%). The time from syncope to test was similar for patients with positive and negative tilt table test results: 28 (1-500) vs 32 (2-700) days (NS). No significant relation was observed between the results of the test and the occurrence of spontaneous syncope during the week, the month or the three months previous to the procedure. However, in men and in patients older than 50 years a higher rate of positive tests was observed if the tilt test was performed within the first month after the last spontaneous syncope. CONCLUSIONS: The time from the last previous spontaneous syncope to the head-up tilt test does not have a significant impact on test outcome in the overall population with suspected neurocardiogenic syncope. However, the rate of positivity might decrease in men and patients older than 50 years if the test is performed later than one month after the spontaneous syncopal episode. PMID- 12892625 TI - [Infective endocarditis in patients with chronic liver disease: clinical and prognostic assessment]. AB - INTRODUCTION AND OBJECTIVES: Chronic liver disease increases the susceptibility to bacterial infections and infective endocarditis. Our aim was to determine the clinical and microbiological features and the prognosis in patients with chronic liver disease who also had infective endocarditis. PATIENTS AND METHOD: One hundred and seventy-four consecutive inpatients at our institution were recruited and followed. Thirty of them had chronic liver disease. Clinical, microbiological and echocardiographic variables were analyzed and, in some cases, histological variables were also recorded. RESULTS: Patients with chronic liver disease were younger (36 11 vs 54 18 years; p < 0.01) and had a larger proportion of intravenous drug users (73 vs 16%; p < 0.01), HIV infection (47 vs 10%; p < 0.01), right valve involvement and spleen enlargement, but heart failure appeared less often (7 vs 34%; p = 0.003). Thirty percent of the patients with and 51% of patients without chronic liver disease underwent surgery for infective endocarditis. Total mortality among patients with and without chronic liver disease was 40% and 31%, respectively. After adjustment for age and for the incidence of congestive heart failure, chronic liver disease doubled mid-term mortality with a RR = 2.45 (p = 0.015). CONCLUSIONS: Chronic liver disease has a significant impact on the prognosis in patients with infective endocarditis, and these patients should therefore be considered a high risk group. PMID- 12892626 TI - [Consensus document for the management of patients with atrial fibrillation in hospital emergency departments]. AB - Atrial fibrillation (AF) is the most prevalent arrhythmia in hospital emergency departments and is a serious disease associated with a twofold increase in morbidity and a high mortality rate. However, the management of AF in this scenario is variable and frequently inadequate. This is probably a consequence of the diverse clinical aspects and therapeutic options to consider in the management of patients with AF. Therefore, implementation of specific, coordinated management strategies by the different care providers involved is needed to improve the quality of care and optimize the use of human and material resources. This document presents the guidelines recommended by the Spanish Society of Cardiology (SEC) and the Spanish Society of Emergency Medicine (SEMES) for the management of AF in hospital emergency departments. These guidelines are based on published scientific evidence and are applicable to most emergency departments in Spain. Specific management strategies are proposed for the conversion and maintenance of sinus rhythm, heart rate control during AF, prophylaxis for thrombi and emboli, and hospital admission and discharge protocols. PMID- 12892627 TI - [Right aortic arch with left aberrant subclavian artery]. PMID- 12892628 TI - [Transition from intravenous to subcutaneous prostacyclin in pulmonary hypertension]. AB - Treatment of arterial pulmonary hypertension with epoprostenol (intravenous prostacyclin) improves survival and quality of life, but the need for an implanted central venous catheter is associated with frequent complications, that often (as in the case of infection or dislodgment) are serious and require catheter replacement. Treprostinil is a prostacyclin analogue suitable for continuous subcutaneous administration. We report the successful transition from intravenous epoprostenol to subcutaneuos treprostinil in four patients with severe pulmonary hypertension who suffered from serious complications associated with the epoprostenol infusion system. PMID- 12892629 TI - [Guided transcatheter valvulotomy in pulmonary atresia with intact ventricular septum]. AB - Transcatheter valvulotomy in pulmonary atresia with an intact ventricular septum can be used as a first step to create biventricular circulation and to stimulate further development of the hypoplastic right ventricle. We describe our experience in a case of a neonate with this congenital cardiac defect who underwent successful transcatheter perforation of the atretic pulmonary valve. This report highlights the utility of a special technique based on the use of a gooseneck snare positioned just above the atretic valve to guide the advance of a coronary guidewire. Other therapeutic alternatives are considered. PMID- 12892630 TI - [Closure of a Blalock-Taussig shunt with an Amplatzer sevice after the Fontan operation]. PMID- 12892631 TI - [Improvement of graft vasculopathy following mycophenolate mofetil administration in a child]. PMID- 12892632 TI - Does endocytosis occur in fungal hyphae? AB - The evidence and arguments for and against the occurrence of endocytosis in fungal hyphae are summarized. The balance of evidence is in favour of the existence of endocytosis. This is supported by an analysis of the recently sequenced Neurospora genome which strongly suggests that this fungus possesses the complex protein machinery required to conduct endocytosis. PMID- 12892634 TI - Functional analysis of mutations in the human carnitine/acylcarnitine translocase in Aspergillus nidulans. AB - Deficiency of the carnitine/acylcarnitine translocase (CACT), the most severe disorder of fatty acid beta-oxidation, is usually lethal in both humans and animals, precluding the development of animal models of the disease. In contrast, CACT deficiency is conditionally lethal in the fungus Aspergillus nidulans, since loss-of-function mutations in acuH, the translocase structural gene, do not prevent growth on carbon sources other than ketogenic compounds, such as fatty acids. Here, we describe the molecular characterization of extant acuH alleles and the development of a fungal model for CACT deficiency based on the ability of human CACT to fully complement, when expressed at physiological levels, the growth defect of an A. nidulans DeltaacuH strain on acetate and long-chain fatty acids. By using growth tests and in vitro assays this model enabled us to carry out a functional characterization of human CACT mutations showing that it may be useful for distinguishing potentially pathogenic human CACT missense mutations from neutral, single residue substitution-causing polymorphisms. PMID- 12892633 TI - Functional and genetic characterization of calmodulin from the dimorphic and pathogenic fungus Paracoccidioides brasiliensis. AB - Calmodulin (CaM) modulates intracellular calcium signalling and acts on several metabolic pathways and gene expression regulation in many eukaryotic organisms including human fungal pathogens, such as Candida albicans and Histoplasma capsulatum. The temperature-dependent dimorphic fungus Paracoccidioides brasiliensis is the aetiological agent of paracoccidioidomycosis (PCM). The mycelium (M) to yeast (Y) transition has been shown to be essential for establishment of the infection, although the precise molecular mechanisms of dimorphism in P. brasiliensis are still unknown. In this work, several inhibitory drugs of the Ca(2+)/calmodulin signalling pathway were tested to verify the role of this pathway in the cellular differentiation process of P. brasiliensis. EGTA and the drugs calmidazolium (R24571), trifluoperazine (TFP), and W7 were able to inhibit the M-Y transition. We have cloned and characterized the calmodulin gene from P. brasiliensis, which comprises 924 nucleotides and five introns that are in a conserved position among calmodulin genes. PMID- 12892635 TI - Entry into the stationary phase is associated with a rapid loss of viability and an apoptotic-like phenotype in the opportunistic pathogen Aspergillus fumigatus. AB - When the opportunistic pathogen Aspergillus fumigatus entered the stationary phase, there was a rapid loss in cell viability which was associated with the appearance of markers characteristic of apoptosis, namely annexin V-FITC binding to the cytoplasmic membrane, demonstrating exposure of phosphatidylserine to the outer leaflet of the membrane; and TUNEL (terminal deoxynucleotidyl transferase mediated dUTP nick end labeling) staining of the nuclei, indicating DNA fragmentation. This was followed later by a loss of membrane integrity as revealed by propidium iodide staining. The development of the apoptotic phenotype was blocked when the protein synthesis inhibitor cycloheximide was added to the culture 1h prior to the onset of the stationary phase, demonstrating active participation of the cell. In addition, intracellular activity against substrates specific for caspase-1 and -8 also increased on stationary phase entry and the development of the apoptotic phenotype was blocked when the cell permeant caspase inhibitor Z-FAD-fmk was present in the medium. Cell death in A. fumigatus during the stationary phase therefore appears to share similarities to apoptotic cell death in higher eukaryotes and to be dependent on a caspase-like activity. PMID- 12892636 TI - Haploid fruiting in Cryptococcus neoformans is not mating type alpha-specific. AB - Under appropriate conditions, haploid Cryptococcus neoformans cells can undergo a morphological switch from a budding yeast form to develop hyphae and viable basidiospores, which resemble those produced by mating. This process, known as haploid fruiting, was previously thought to occur only in MATalpha strains. We identified two new strains of C. neoformans var. neoformans serotype D that are MATa type and are able to haploid fruit. Further, a MATa reference strain, B 3502, also produced hyphae and fruited after prolonged incubation on filament agar. Over-expression of STE12a dramatically enhanced the ability of all MATa strains tested to filament. Segregation analysis of haploid fruiting ability confirmed that haploid fruiting is not MATalpha-specific. Our results indicate that MATa cells are intrinsically able to haploid fruit and previous observations that they do not were probably biased by the examination of a small number of genetically related isolates that have been maintained in the laboratory for many years. PMID- 12892637 TI - Single-strand-conformation polymorphism of ribosomal DNA for rapid species differentiation in genus Phytophthora. AB - Single-strand-conformation polymorphism (SSCP) of ribosomal DNA of 29 species (282 isolates) of Phytophthora was characterized in this study. Phytophthora boehmeriae, Phytophthora botryosa, Phytophthora cactorum, Phytophthora cambivora, Phytophthora capsici, Phytophthora cinnamomi, Phytophthora colocasiae, Phytophthora fragariae, Phytophthora heveae, Phytophthora hibernalis, Phytophthora ilicis, Phytophthora infestans, Phytophthora katsurae, Phytophthora lateralis, Phytophthora meadii, Phytophthora medicaginis, Phytophthora megakarya, Phytophthora nicotianae, Phytophthora palmivora, Phytophthora phaseoli, Phytophthora pseudotsugae, Phytophthora sojae, Phytophthora syringae, and Phytophthora tropicalis each showed a unique SSCP pattern. Phytophthora citricola, Phytophthora citrophthora, Phytophthora cryptogea, Phytophthora drechsleri, and Phytophthora megasperma each had more than one distinct pattern. A single-stranded DNA ladder also was developed, which facilitates comparison of SSCP patterns within and between gels. With a single DNA fingerprint, 277 isolates of Phytophthora recovered from irrigation water and plant tissues in Virginia were all correctly identified into eight species at substantially reduced time, labor, and cost. The SSCP analysis presented in this work will aid in studies on taxonomy, genetics, and ecology of the genus Phytophthora. PMID- 12892638 TI - Characterization of the genomic organization of the region bordering the centromere of chromosome V of Podospora anserina by direct sequencing. AB - A Podospora anserina BAC library of 4800 clones has been constructed in the vector pBHYG allowing direct selection in fungi. Screening of the BAC collection for centromeric sequences of chromosome V allowed the recovery of clones localized on either sides of the centromere, but no BAC clone was found to contain the centromere. Seven BAC clones containing 322,195 and 156,244bp from either sides of the centromeric region were sequenced and annotated. One 5S rRNA gene, 5 tRNA genes, and 163 putative coding sequences (CDS) were identified. Among these, only six CDS seem specific to P. anserina. The gene density in the centromeric region is approximately one gene every 2.8kb. Extrapolation of this gene density to the whole genome of P. anserina suggests that the genome contains about 11,000 genes. Synteny analyses between P. anserina and Neurospora crassa show that co-linearity extends at the most to a few genes, suggesting rapid genome rearrangements between these two species. PMID- 12892639 TI - Investigating the role of a Verticillium fungicola beta-1,6-glucanase during infection of Agaricus bisporus using targeted gene disruption. AB - Studies on the mycopathogen Verticillium fungicola have shown the up-regulation of beta-1,6-glucanases when grown in the presence of host cell walls and host cell wall components including chitin. These cell-wall-degrading enzymes are hypothesized to contribute to the pathogenic ability of mycopathogens. A beta-1,6 glucanase gene, VfGlu1, showing high similarity to beta-1,6-glucanase genes from Hypocrea virens, Neotyphodium sp., and Trichoderma harzianum, was isolated using degenerate PCR from V. fungicola, a serious mycopathogen of the cultivated mushroom Agaricus bisporus. Agrobacterium-mediated transformation of V. fungicola using homologous DNA from VfGlu1 resulted in homologous integration at the VfGlu1 locus in 75% of transformants, generating mutants disrupted in the VfGlu1 gene. VfGlu1 mutants displayed reduced virulence and diminished ability to utilize chitin as a carbon source, implicating VfGlu1 in the disease process. Agrobacterium-mediated transformation affords an efficient technique for the disruption of genes associated with disease symptom development in the complex V. fungicola-A. bisporus interaction. PMID- 12892640 TI - Molecular cloning, characterization, and expression of a cDNA encoding an endochitinase gene from the mycoparasite Stachybotrys elegans. AB - Stachybotrys elegans is a mycoparasite of the soilborne plant pathogenic fungus Rhizoctonia solani. The mycoparasitic activity of S. elegans is correlated with the production of cell wall degrading enzymes such as chitinases. This report details the cloning by RACE-PCR and characterization of a full-length cDNA clone, sechi44, that appears to encode an extracellular endochitinase. An analysis of the sechi44 sequence indicates that this gene contains a 1269-bp ORF and encodes a 423-aa polypeptide. The SECHI44 protein has a calculated molecular weight of 44.1kDa and pI of 5.53. Since the SECHI44 protein also appears to encode a signal peptide, an extracellular location for the corresponding protein is predicted. Comparison of SECHI44 sequence with known sequences of fungal endochitinases revealed that SECHI44 is grouped with endochitinases from other mycoparasites. Real-time quantitative RT-PCR analysis showed an elevated level of expression of sechi44 (21-fold) in chitin-rich (induced) as compared to no-carbon (non-induced) culture conditions. In dual culture, the temporal expression of sechi44 increased after 2 days of contact with R. solani, reaching a 10-fold increase after 9 days, followed by a decrease to basic expression level at 12 days. Interestingly, inhibition of sechi44 expression was observed when S. elegans hyphae were in close proximity with R. solani hyphae. PMID- 12892641 TI - The pH-induced glycosylation of secreted phosphatases is mediated in Aspergillus nidulans by the regulatory gene pacC-dependent pathway. AB - In this communication, we show that the pacC(c)14 mutation drastically reduced the mannose and N-acetylglycosamine content of the pacA-encoded acid phosphatase secreted by the fungus Aspergillus nidulans when grown at 22 degrees C, pH 5.0, compared to a control strain. The staining after PAGE was not observed for the pacA-encoded acid phosphatase, while the palD-encoded Pi-repressible alkaline phosphatase had an altered electrophoretic mobility. In addition, the secreted acid phosphatase also had a reduced number of isoforms visualized by staining after IEF and glycosylation had a protective effect against its heat inactivation. We also show that a full-length version of gene pacC-1 cloned from Neurospora crassa complemented the pacC(c)14 mutation of A. nidulans, including the remediation of both the acid and alkaline Pi-repressible phosphatases secreted at pH 5.0, which indicates that glycosylation of secreted phosphatases is mediated in A. nidulans by the conserved PacC pathway that governs pH responsive gene expression. PMID- 12892642 TI - Transcriptional control of neuronal diversification in the retina. AB - During embryonic development, the array of vastly different neuronal types that are incorporated into the functional architecture of the mature neuroretina derives from a common population of multipotent retinal progenitor cells (RPCs). Retinogenesis proceeds in a precise chronological order, with the seven principal cell classes generated in successive phases. Cell biological experiments established that this histogenetic order, at least in part, reflects intrinsic changes within the RPC pool. In recent years a number of molecules controlling various aspects of cell fate specification from RPCs have been identified. However, few attempts have been made to integrate previous concepts that emerged from cell biological studies and more recent results based on molecular genetic experiments. This review aims at providing an overview of recent advances in our understanding of the cellular and molecular mechanisms underlying retinal neuronal diversification, with a particular focus on cell-intrinsic factors. PMID- 12892643 TI - The assessment of L- and M-cone specific electroretinographical signals in the normal and abnormal human retina. AB - Electroretinography (ERG) is a non-invasive method that can contribute to a description of the functional organization of the human retina under normal and pathological circumstances. The physiological and pathophysiological processes leading to an ERG signal can be better understood when the cellular origins of the ERG are identified. The ERG signal recorded at the cornea is initiated by light absorption in the photoreceptors which leads to activity in the photoreceptors and in their post-receptoral pathways. Light absorption in distinct photoreceptor types may lead to different ERG responses caused either by differences between the photoreceptors or between their post-receptoral pathways. The description of contributions of the different photoreceptor types to the ERG may therefore give more detailed insight in the origins of the ERG. Such a description can be obtained by isolating the responses of a single photoreceptor type. Nowadays, careful control of differently colored light sources together with the relatively well-known cone and rod fundamentals enables a precise description and control of photoreceptor excitation. Theoretically, any desired combination of photoreceptor excitation modulation can be achieved, including conditions in which the activity in only one photoreceptor type is modulated (silent substitution). In this manner the response of one photoreceptor type is isolated without changing the state of adaptation. This stimulus technique has been used to study the contribution of signals originating in the different photoreceptor types to the human ERG. Furthermore, by stimulating two or more photoreceptor types simultaneously, the interaction between the different signals can be studied. With these new techniques results of measurements in healthy subjects and patients with retinal diseases can be compared. This approach should ultimately help to develop better diagnostic tools and result in a fuller description of the changes and the pathophysiological mechanisms in retinal disorder. Finally, data obtained with cone and rod specific stimuli may lead to a reinterpretation of the standard ERG used in a clinical setting. PMID- 12892644 TI - Neural remodeling in retinal degeneration. AB - Mammalian retinal degenerations initiated by gene defects in rods, cones or the retinal pigmented epithelium (RPE) often trigger loss of the sensory retina, effectively leaving the neural retina deafferented. The neural retina responds to this challenge by remodeling, first by subtle changes in neuronal structure and later by large-scale reorganization. Retinal degenerations in the mammalian retina generally progress through three phases. Phase 1 initiates with expression of a primary insult, followed by phase 2 photoreceptor death that ablates the sensory retina via initial photoreceptor stress, phenotype deconstruction, irreversible stress and cell death, including bystander effects or loss of trophic support. The loss of cones heralds phase 3: a protracted period of global remodeling of the remnant neural retina. Remodeling resembles the responses of many CNS assemblies to deafferentation or trauma, and includes neuronal cell death, neuronal and glial migration, elaboration of new neurites and synapses, rewiring of retinal circuits, glial hypertrophy and the evolution of a fibrotic glial seal that isolates the remnant neural retina from the surviving RPE and choroid. In early phase 2, stressed photoreceptors sprout anomalous neurites that often reach the inner plexiform and ganglion cell layers. As death of rods and cones progresses, bipolar and horizontal cells are deafferented and retract most of their dendrites. Horizontal cells develop anomalous axonal processes and dendritic stalks that enter the inner plexiform layer. Dendrite truncation in rod bipolar cells is accompanied by revision of their macromolecular phenotype, including the loss of functioning mGluR6 transduction. After ablation of the sensory retina, Muller cells increase intermediate filament synthesis, forming a dense fibrotic layer in the remnant subretinal space. This layer invests the remnant retina and seals it from access via the choroidal route. Evidence of bipolar cell death begins in phase 1 or 2 in some animal models, but depletion of all neuronal classes is evident in phase 3. As remodeling progresses over months and years, more neurons are lost and patches of the ganglion cell layer can become depleted. Some survivor neurons of all classes elaborate new neurites, many of which form fascicles that travel hundreds of microns through the retina, often beneath the distal glial seal. These and other processes form new synaptic microneuromas in the remnant inner nuclear layer as well as cryptic connections throughout the retina. Remodeling activity peaks at mid-phase 3, where neuronal somas actively migrate on glial surfaces. Some amacrine and bipolar cells move into the former ganglion cell layer while other amacrine cells are everted through the inner nuclear layer to the glial seal. Remodeled retinas engage in anomalous self-signaling via rewired circuits that might not support vision even if they could be driven anew by cellular or bionic agents. We propose that survivor neurons actively seek excitation as sources of homeostatic Ca(2+) fluxes. In late phase 3, neuron loss continues and the retina becomes increasingly glial in composition. Retinal remodeling is not plasticity, but represents the invocation of mechanisms resembling developmental and CNS plasticities. Together, neuronal remodeling and the formation of the glial seal may abrogate many cellular and bionic rescue strategies. However, survivor neurons appear to be stable, healthy, active cells and given the evidence of their reactivity to deafferentation, it may be possible to influence their emergent rewiring and migration habits. PMID- 12892645 TI - Redox regulation in the lens. AB - The high content of glutathione (GSH) in the lens is believed to protect thiols in structural proteins and enzymes for proper biological functions. The lens has both biosynthetic and regenerating systems for GSH to maintain its large pool size. However, ageing lenses or lenses under oxidative stress show an extensively diminished size of GSH pool with some protein thiols being S-thiolated by oxidized non-protein thiols to form protein-thiol mixed disulfides, either as protein-S-S-glutathione (PSSG) or protein-S-S-cysteine (PSSC) or protein-S-S gamma-glutamylcysteine. It was shown in an H(2)O(2)-induced cataract model that PSSG formation precedes a cascade of events before cataract formation, starting with protein disulfide crosslinks, protein solubility loss and high molecular weight aggregation. Furthermore, this early oxidative damage in protein thiols can be spontaneously reversed in H(2)O(2) pretreated lenses if the oxidant is removed in time. This dethiolation process appears to have mediated through a redox-regulating enzyme, thioltransferase (TTase), which is ubiquitously present in microbial, plant and animal tissues, including the lens. The GSH-dependent, low molecular weight (11.8 kDa) cytosolic enzyme plays an important role in oxidative defense and can modulate key metabolic enzymes in the glycolytic pathway. The enzyme repairs oxidatively damaged proteins/enzymes through its unique catalytic site with a vicinal cysteine moiety, which can specifically dethiolate protein-S-S-glutathione and restore protein free SH groups for proper enzymatic or protein functions. Most importantly, it has been demonstrated that thioltransferase has a remarkable resistance to oxidation (H(2)O(2)) in cultured human and rabbit lens epithelial cells under oxidative stress conditions when other oxidation defense systems of GSH peroxidase and GSH reductase are severely inactivated. A second repair enzyme, thioredoxin (TRx), which is NADPH-dependent, is widely found in many lower and higher life forms of life. It can dethiolate protein disulfides and thus is an extremely important regulator for redox homeostasis in the cells. Thioredoxin has been recently found in the lens and has been shown to participate in the repair process of oxidatively damaged lens proteins/enzymes. These two enzymes may work synergistically to regulate and repair thiols in lens proteins and enzymes, keeping a balanced redox potential to maintain the function of the lens. PMID- 12892646 TI - Vitamin A metabolism in the retinal pigment epithelium: genes, mutations, and diseases. AB - Mutations in the genes necessary for the metabolism of vitamin A (all-trans retinol) and cycling of retinoids between the photoreceptors and retinal pigment epithelium (RPE) (the visual cycle) have recently emerged as an important class of genetic defects responsible for retinal dystrophies and dysfunctions. Research into the causes and treatment of diseases resulting from defects in retinal vitamin A metabolism is currently the subject of intense interest, since disorders affecting the RPE are, in principle, more accessible to therapeutic intervention than those affecting the proteins of photoreceptor cells. This chapter presents an overview of the visual cycle, as well as the function of the RPE genes involved in the conversion of vitamin A to 11-cis retinal, the chromophore of the visual pigments. The identification of disease-causing mutations in this group of genes is described as well as the associated phenotypes that range from stationary night blindness to childhood-onset severe visual handicap. Consideration is also given to alternative genetic paradigms potentially relevant to defects in vitamin A metabolism, including a discussion of the relationship of this pathway to age-related macular degeneration, a non Mendelian disease of late onset. Finally, progress and prospects for targeted therapeutic intervention in vitamin A metabolism are presented, including retinoid and gene replacement therapy. On the basis of early successes in animal models, and plans underway for Phase I/II clinical trials, it is hoped that the near future will bring effective therapies for many retinal dystrophy patients with defects in vitamin A metabolism. PMID- 12892647 TI - Cortical alpha-adrenoceptor downregulation by tricyclic antidepressants in the rat brain. AB - The aim of the present study was to examine the effect of chronic tricyclic antidepressants (TCAs) treatment on the density of alpha-adrenoceptors in the rat brain. Density of alpha1- and alpha2-adrenoceptors was measured in cortex and hippocampus of rats treated with imipramine (IMI, 5mg/kg body weight), desipramine (DMI, 10mg/kg body weight), clomipramine (CMI, 10mg/kg body weight) and amitriptyline (AMI, 10mg/kg body weight), for 40 days, using [3H]prazosin and [3H]clonidine, respectively. The density of cortical alpha1-adrenoceptors was significantly decreased with IMI (46%), DMI (21%), CMI (50%) and AMI (67%) treatment, without altering the affinity of the receptor. The density of cortical alpha2-adrenoceptors was also significantly decreased with DMI (69%), CMI (81%) and AMI (80%) treatment, without affecting the affinity for [3H]clonidine. The density of hippocampal alpha1-adrenoceptors was significantly decreased only with AMI treatment (47%), without affecting the affinity for [3H]prazosin. However, no change in hippocampal alpha2-adrenoceptor density was observed with any of these TCAs. The results suggest that chronic antidepressant (AD) treatment downregulates the cortical, but not hippocampal, alpha1- and alpha2-adrenoceptors in rat brain. The region-specific downregulation of alpha1- and alpha2 adrenoceptors density, which occur after prolonged AD treatment, may underline the therapeutic mechanism of action. PMID- 12892648 TI - Effects of desipramine on regional serotonin synthesis in the rat brain: acute and chronic autoradiographic studies. AB - Various studies have implicated the involvement of noradrenaline (NA) and/or serotonin (5-hydroxytryptamine (5-HT)) in the pathogenesis and treatment of depression. The aim of the present study was to investigate the effects of acute and 7 days of administration of desipramine, a NA re-uptake inhibitor, on the rate of 5-HT synthesis in the rat brain. The study was done by an autoradiographic method using alpha-[14C]-methyl-L-tryptophan as a tracer. The acute (10mg/kg, i.p., 2h before i.v. infusion of the tracer) or 7 days of desipramine (10mg/kg per day, i.p.) did not affect plasma tryptophan (Trp) concentrations, as compared to control (saline treated) rats. Acute treatment with desipramine decreased the rate of 5-HT synthesis in the brain regions that contain 5-HT cell bodies between 19 and 28%, and increased the rate of 5-HT synthesis in the majority of areas containing 5-HT terminals between 21 and 65%. In contrast to the acute treatment, a 7-day administration increased 5-HT synthesis rates in the dorsal raphe (24%), but decreased it in raphe magnus (35%), superior olive (45%), caudate (31%), superior (38%) and inferior (53%) colliculus, and in the auditory cortex (35%). This suggests that the effect of desipramine on 5-HT synthesis rate is time-dependent and differs in the cell bodies and structures containing 5-HT nerve terminals. PMID- 12892649 TI - Extracellular adenine nucleotides metabolism in astrocyte cultures from different brain regions. AB - Primary astrocyte cultures from hippocampus, cortex and cerebellum presented different extracellular pattern of adenine nucleotide hydrolysis. The ATP/ADP hydrolysis ratio was 8:1 for hippocampal and cortical astrocytes and 5:1 for cerebellar astrocytes. The AMP hydrolysis in cerebellar astrocytes was seven-fold higher than in cortical or hippocampal cells. No accumulation of extracellular adenosine in all structures studied was observed. Dipyridamol increased significantly inosine levels in the extracellular medium of hippocampal and cortical, but not in cerebellar astrocytes medium. A higher expression of ecto-5' nucleotidase was identified by RT-PCR in cerebellum. The differences observed may indicate functional heterogeneity of nucleotides in the brain. PMID- 12892650 TI - The pentylenetetrazole-kindling model of epilepsy in SAMP8 mice: glial-neuronal metabolic interactions. AB - Recently, a new experimental model of epilepsy was introduced by the authors [Neurochem. Int. 40 (2002) 413]. This model combines pentylenetetrazole (PTZ) kindling in senescence-accelerated mice P8 (SAMP8), a genetic model of aging. Since imbalance of glutamate and GABA is a major cause of seizures, the study of glial-neuronal interactions is of primary importance. Nuclear magnetic resonance spectroscopy (NMRS) is an excellent tool for metabolic studies. Thus, we examined whether NMRS when combined with administration of [1-13C]glucose and [1,2 13C]acetate might give valuable insights into neurotransmitter metabolism in this new model of epilepsy and aging. The 2- and 8-month-old SAMP8 were kindled with PTZ alone, received PTZ and phenobarbital (PB), or served as controls. In older animals, PTZ-kindling decreased labeling in glutamate C-4 from [1-13C]glucose, whereas, in the younger mice, labeling in glutamine C-4 was decreased both from [1-13C]glucose and [1,2-13C]acetate. It could be concluded that PTZ-kindling affected astrocytes in younger and glutamatergic neurons in older animals. In the presence of PTZ, phenobarbital decreased labeling of most metabolites in all cell types, except GABAergic neurons, from both labeled precursors in the younger animals. However, in older animals only GABAergic neurons were affected by phenobarbital as indicated by an increase in GABA labeling. PMID- 12892651 TI - Gene expression of glutamate receptors GluR1 and NR1 is differentially modulated in striatal neurons in rats after 6-hydroxydopamine lesion. AB - In the present study, we attempted to address the modulation of the gene expression of N-methyl-D-aspartate (NMDA) and alpha-amino-3-hydroxy-5-methyl-4 isoxazole-propionate (AMPA) glutamate receptors in the neostriatum of the 6 hydroxydopamine (6-OHDA)-lesioned rat, an animal model of Parkinson's disease. After 2 weeks of lesion, reverse transcriptase-polymerase chain reactions (RT PCRs) revealed significant reduction in GluR1 mRNA expression but a significant enhancement of NR1 mRNA expression in the striatal tissues of the lesioned side. No modulation in the mRNA expression of GluR2, GluR3, GluR4 and NR2B were found. Immunofluorescence with digital imaging analysis also demonstrated a significant reduction in GluR1 immunoreactivity in the lesioned neostriatum. Interestingly, the reduction in GluR1 immunoreactivity was primarily observed in presumed striatal medium spiny neurons but not in parvalbumin-labeled striatal GABAergic interneurons. Immunoreactivity for GluR2, GluR2/3, GluR4, NR1 and NR2B was unchanged in neurons of the neostriatum of the lesioned side. The present results indicate that there is an opposite trend in modulation in the gene expressions of GluR1 and NR1 in the neostriatum of 6-OHDA-lesioned rats after dopamine denervation. Modulation of GluR1 mRNA and immunoreactivity is likely to be limited in the striatal projection neurons. These findings have implications for the use of NMDA and AMPA receptor antagonists in the treatment of Parkinson's disease. PMID- 12892652 TI - Fasting plasma ghrelin levels in subtypes of anorexia nervosa. AB - Ghrelin has a role in regulating eating behavior and energy metabolism in the central nervous system, and has been reported to play an important role in the pathophysiology of anorexia nervosa (AN). The aim of the present study was to compare fasting plasma ghrelin levels in different subtypes of untreated AN patients. The subjects included 39 female AN patients and 11 female controls. The patients were then divided into two subtypes as follows: 19 AN patients with restricting (AN-R) and 20 AN patients with binge-eating/purging (AN-BP) form of the illness. Blood samples from subjects after an overnight fast were used to analyze plasma ghrelin concentrations. Plasma ghrelin concentrations in both AN-R and AN-BP were negatively correlated with body mass index (BMI). The mean plasma ghrelin levels in both AN-R and AN-BP were significantly higher than that in controls. The mean ghrelin level in AN-BP was significantly higher than that in AN-R. However, mean BMI and serum potassium in both groups were not significantly different. These results suggest that both BMI and the presence of binge eating/purging may have some influence on fasting plasma ghrelin levels in patients with AN. PMID- 12892653 TI - Neuroendocrine alterations in a high-dose, extended-access rat self administration model of escalating cocaine use. AB - One approach for studying cocaine addiction has been to permit escalating patterns of self-administration (SA) by rats by prolonging daily drug availability. Rats provided long access (LgA) to high cocaine doses, but not rats provided shorter cocaine access (ShA), progressively escalate their cocaine intake and display characteristics of human addiction. The purpose of the present study was to investigate the effects of 14 days of ShA or LgA, high-dose cocaine SA on plasma corticosterone (CORT), prolactin (PRL), and related mRNAs. Acutely, cocaine SA increased plasma CORT and reduced plasma PRL levels. SA training produced circadian increases in CORT that appeared to occur in anticipation of cocaine availability. With repeated LgA, high-dose SA, the daily CORT area under the curve (AUC) progressively decreased, apparently due to tolerance to cocaine's effects on CORT and a reduction in basal CORT levels. In contrast, the daily CORT AUC in ShA rats increased across testing despite constant rates of SA. When measured 12 days after SA testing, pro-opioimelanocortin and glucocorticoid receptor mRNA levels in the anterior pituitary were lower in LgA rats than in ShA rats. The effects of SA on PRL remained constant across SA testing in LgA rats, but increased in duration in ShA rats. Anterior pituitary dopamine D2 receptor mRNA levels were lower in LgA rats than in ShA rats. These findings indicate that the transition to escalating patterns of SA may be associated with altered levels of hormones and gene expression within neuroendocrine systems. Such changes may underlie the onset of human addictive disease. PMID- 12892654 TI - Andropause and psychopathology: minor symptoms rather than pathological ones. AB - This study examined the psychological symptomatology of men diagnosed with andropause and the association between calculated free testosterone (T) and depressed mood, anxiety and quality of life. Subjects were 153 men, aged 50-70 years, who participated in a screening of andropause. Total testosterone, FSH, LH and SHBG levels were measured. Depressed mood was assessed with the Carroll Rating Scale, anxiety with the "anxiety-insomnia" dimension of the General Health Questionnaire, and quality of life with the World Health Organisation Quality of Life questionnaire. The results showed that levels of free T decreased with age, whereas FSH and LH increased. Carroll Rating Scale scores were higher among hypogonadal subjects, but the mean score was low and not pathological. A negative correlation was observed between severity of depression as assessed by the Carroll Rating Scale and free T levels. However, subjects with a significant score on this scale did not exhibit different free T levels compared to subjects with a non-significant depressive score. Anxiety and quality of life did not differ between hypogonadal and eugonadal subjects. The present study therefore suggests that andropause is not characterised by specific psychological symptoms, but may be associated with "depressive symptoms" that are not considered as pathological. PMID- 12892655 TI - Serotonergic activation stimulates the pituitary-adrenal axis and alters interleukin-1 mRNA expression in rat brain. AB - Interactions between neurotransmitters and immunomodulators within the central nervous system may be functionally relevant for communication between the immune system and the brain. Previous studies indicate that cytokines such as interleukin-1 (IL-1) alter activity of the serotonergic system at multiple levels. This study tested the hypothesis that serotonergic activation modulates cytokine mRNA expression in brain. Serotonergic activation was induced by injecting rats intraperitoneally (i.p.) prior to dark onset with the serotonin precursor L-5-hydroxytryptophan (5-HTP; 100 mg/kg). Cytokine mRNA expression in discrete brain regions at selected time points was determined by means of ribonuclease protection assay. Plasma corticosterone concentrations were also measured to determine if the hypothalamic-pituitary-adrenal axis is activated in response to this treatment, which potentially could exert feedback regulating cytokine message expression in brain. Plasma corticosterone was elevated for 4 h after 5-HTP administration. At this time IL-1alpha mRNA expression was reduced in the hippocampus, hypothalamus, and brainstem, and IL-1beta mRNA was reduced in the hippocampus. Six hours after 5-HTP injection, IL-1beta mRNA increased in the hypothalamus. These results show that activation of the serotonergic system affects cytokine message expression in rat brain, possibly by actions of corticosterone. PMID- 12892656 TI - Estradiol selectively affects processing of conspecifics' faces in female rhesus monkeys. AB - Estrogen deficiency following ovariectomy or menopause increases the risk of developing diseases such as osteoporosis and may also lead to memory impairment. Although estrogen replacement therapy (ERT) alleviates many symptoms associated with estrogen loss, it is not clear whether it also benefits cognitive function. The effect of estrogens upon cognition can best be studied in an animal model of human menopause, in which estrogen levels can be experimentally manipulated. Six young ovariectomized female rhesus monkeys (6-9 years old) were tested on a battery of touchscreen-based cognitive tasks, including the Matching-to-Sample (MTS) task with mixed delays and the spatial, object, and face conditions of the Delayed Recognition Span Test (DRST). Monkeys were tested 5 days a week, one task per week, for a total of 8 months, while undergoing treatments with placebo and ethinyl estradiol (EE2) in alternating 28-days blocks. Blood samples were collected to verify EE2 levels. We also observed the monkeys by video monitor during test sessions and recorded locomotor activity and response topology. Performance on the face-DRST, a task that involved selecting the new face in an increasing array of rhesus monkey faces, was impaired by EE2 treatment, as compared to placebo. Other tasks were unaffected by EE2. There was no clear evidence of EE2 effects upon motor activity or anxiety. In order to test the reliability of our findings, we conducted an additional experiment in which the monkeys were again given the face-DRST with different categories of face stimuli for 4 months, while receiving placebo and EE2 in alternating 7-days blocks. They performed each task 4-5 days/week for 4 weeks with (1) the same rhesus monkey faces as in the first experiment, (2) human faces, (3) chimpanzee faces, and (4) novel rhesus monkey faces. Face-DRST performance did not vary as a function of treatment when human or chimpanzee faces were used as stimuli. In contrast, periods of EE2 treatment were associated with a lower performance for both sets of rhesus monkey faces. These findings suggest that EE2 treatment has a detrimental effect on processing faces of conspecifics by female rhesus monkeys. We speculate that estrogens may produce this effect by enhancing emotional reactivity to socially relevant stimuli. PMID- 12892657 TI - Sex-sensitive cognitive performance in untreated patients with early onset gender identity disorder. AB - BACKGROUND: We explored whether the cognitive performance of gender identity disorder patients (GID) was comparable to that of their biological sex or skewed towards that of their gender identity. METHOD: We tested four potentially sex sensitive cognitive factors (rotation, visualization, perception, and verbalization) as well as two neutral factors (logic and arithmetic) in GID patients from Norway (GID-N, n = 33) or the USA (GID-US, n = 19) and in a control group (C, n = 29). The testing was undertaken prior to cross sex hormone treatment. Four-way ANOVA was applied in the final analysis of the cognitive performance and its dependency on different predictors (age, biological sex, education, group). RESULTS: In both GID groups as well as in the control group (C) males excelled in visualization and rotation, also when controlling for potential confounders (biological sex, group, age and education). No female advantage was detected. Furthermore, no interaction between biological sex and group assignment was revealed in the samples. CONCLUSION: In this study the cognitive pattern of GID patients is consistent with that of their biological sex and not that of their gender identity. PMID- 12892658 TI - Two formulas for computation of the area under the curve represent measures of total hormone concentration versus time-dependent change. AB - Study protocols in endocrinological research and the neurosciences often employ repeated measurements over time to record changes in physiological or endocrinological variables. While it is desirable to acquire repeated measurements for finding individual and group differences with regard to response time and duration, the amount of data gathered often represents a problem for the statistical analysis. When trying to detect possible associations between repeated measures and other variables, the area under the curve (AUC) is routinely used to incorporate multiple time points. However, formulas for computation of the AUC are not standardized across laboratories, and existing differences are usually not presented when discussing results, thus causing possible variability, or incompatibility of findings between research groups. In this paper, two formulas for calculation of the area under the curve are presented, which are derived from the trapezoid formula. These formulas are termed 'Area under the curve with respect to increase' (AUCI) and 'Area under the curve with respect to ground' (AUCG). The different information that can be derived from repeated measurements with these two formulas is exemplified using artificial and real data from recent studies of the authors. It is shown that depending on which formula is used, different associations with other variables may emerge. Consequently, it is recommended to employ both formulas when analyzing data sets with repeated measures. PMID- 12892659 TI - Association of testosterone and dihydrotestosterone with externalizing behavior in adolescent boys and girls. AB - BACKGROUND: While an association between androgens and different types of aggression has been well documented in male offenders, the influence of androgens on externalizing behavior in adolescents at risk for antisocial behavior has not been investigated so far. METHODS: Plasma levels of the main androgen metabolites testosterone (T) and 5alpha-dihydrotestosterone (DHT) were measured in N = 87 fourteen-year-old (36 boys, 51 girls) from a prospective longitudinal study of children at risk. Externalizing behavior at age 8, 11 and 14 was assessed using the Achenbach Child Behavior Checklist (CBCL) and Teacher Report Form (TRF). RESULTS: Significant higher androgen levels (T, DHT) were found in male, but not in female adolescents with elevated scores of externalizing behavior. Moreover, boys with persistent externalizing behavior exhibited the highest levels of plasma androgens. CONCLUSIONS: There is a link between T, DHT and externalizing behavior in male adolescents at risk for psychopathology. Due to the findings of highest androgen levels in boys with persistent externalizing behavior, a role of androgens in the development of disruptive or later antisocial disorders can be hypothesized. PMID- 12892660 TI - Attentionally modulated effects of cortisol and mood on memory for emotional faces in healthy young males. AB - Heightened cortisol levels due to stress or acute administration seem to enhance memory for emotional material, independently of emotional valence. An arousal driven neurobiological mechanism involving the amygdala has been proposed. The relation between pre-task salivary measures of cortisol (by convention named 'basal levels') and emotionally modulated memory has not been investigated yet. Given the association between higher basal levels of cortisol and indices of low mood, valence-specific effects on emotionally modulated memory could be expected (e.g. mood-congruent or stimulus-specific forms of processing). This study was designed to investigate the relationship between basal levels of salivary cortisol, self-reported mood and spatial memory for neutral, happy and angry facial expressions in healthy young volunteers (N=31). Memory performance was indexed using a modified version of a computerized object-relocation task, using emotional facial expressions as stimuli. Results showed a significant relation between cortisol and depressive mood. More importantly, both the levels of cortisol and depressive mood were inversely related to the memory performance for the happy facial expressions, while a similar relationship between cortisol and memory performance on angry faces neared significance. An explanation in terms of the down-regulation of social behavior by elevated basal cortisol levels is postulated. PMID- 12892661 TI - Competition kinetics using the UV/H2O2 process: a structure reactivity correlation for the rate constants of hydroxyl radicals toward nitroaromatic compounds. AB - The rate constants for hydroxyl radical reaction toward a set of nitroaromatic substrates kS, have been measured at 25 degrees C using competition experiments in the UV/H2O2 process. For a given pair of substrates S1 and S2, the relative reactivity beta (defined as kS1/kS2) was calculated from the slope of the corresponding double logarithmic plot, i.e., of ln[S1] vs. ln[S2]. This method is more accurate and remained linear for larger conversions in comparison with the plots of ln[S1] and ln[S2] against time. The rate constants measured ranged from 0.33 to 8.6 x 10(9) M(-1)s(-1). A quantitative structure-reactivity relationship was found using the Hammett equation. Assuming sigma values to be additive, a value of -0.60 was obtained for the reaction constant rho. This value agrees with the high reactivity and the electrophilic nature of HO* radical. PMID- 12892662 TI - Remediation of contaminated soil by a solvent/surfactant system. AB - This study investigates a new approach using a solvent/surfactant-aided soil washing process to improve the performance of conventional surfactant-aided soil remediation. Three surfactants (Brij 35, Tween 80, and SDS) and three organic solvents (acetone, triethylamine, and squalane) were used to evaluate the desorption performances of 4,4'-dichlorobiphenyl (DCB) out of three soils with different sorption characteristics. The performance improvement is likely due to better dissolution of the hydrophobic contaminants from the soil assisted by the solvent, and the formation of solvent-incorporated surfactant micelles, which increases both the size (i.e. capacity) and affinity of micelles for more effective contaminant extraction. The foc of soils were found to be important in determining the performance of a solvent/surfactant-aided soil-washing process. Judging from the experimental data and as verified by the two constants in the proposed soil-washing model, as the organic solvent is coexisting with the surfactant micelles, both the marginal soil-washing performance (right after the use of a very small amount of solvent compared to that of none) and the final soil-washing capacity are increased compared to those of a pure surfactant-aided washing process. PMID- 12892663 TI - Modeling VOCs adsorption onto activated carbon. AB - The activated carbon adsorption process is affected by the characteristics of adsorbent, adsorbate and environmental conditions. In this study, both adsorption and desorption processes are assumed to occur simultaneously and a numerical model was developed with a non-linear driving force in conjunction with the Langmuir model for predicting the overall adsorption process. The numerical model provides both adsorption and desorption rate constants and activation energies. The resultant equilibrium constants are of the same order of magnitude as reported by other studies. Results show that the model could well predict the adsorption isotherms and breakthrough curves under various conditions. PMID- 12892664 TI - Characterization of atmospheric PM10 and related chemical species in southern Taiwan during the episode days. AB - The concentrations of atmospheric PM10 on days with episodes of pollution were examined at four different sampling sites (CC, DL, LY, and HK) in southern Taiwan. The related to particulates water-soluble ionic species (Na+, K+, Mg2+, Ca2+, NH4+, Cl-, NO3-, SO4(2-)), carbonaceous species (EC and OC) and metallic species (Zn, Ni, Pb, Fe, Mn, Al, Si, V) were also analyzed. On the episode days of this study, the PM10 mass concentration ranged from 155 to 210 microgm(-3), from 150 to 208 microgm(-3), from 182 to 249 microgm(-3), and from 166 to 228 microgm(-3) at CC, DL, LY, and HK, respectively. The results indicate that the dominant water-soluble species were SO4(2-), NO3-, NH4+, and Cl- at the four sampling sites on these days. Moreover, the high sulfate and nitrate conversion values (SOR and NOR) presented herein suggest that secondary formations from SO2 to SO4(2-) and from NO2 to NO3- are present in significant quantities in the atmosphere of southern Taiwan on episode days. In particular, high SOR and NOR verified that both SO4(2-) and NO3- dominated the increase of atmospheric PM10 concentration in southern Taiwan on episode days. PMID- 12892665 TI - Ammonia inhibition on thermophilic anaerobic digestion. AB - This study evaluated both chronic and acute toxicity of ammonia in thermophilic anaerobic digestion of synthetic wastewater over a range of acclimation concentrations. The inhibition effects of ammonia, in terms of total ammonia nitrogen (TAN), under various pH values and acclimation conditions on thermophilic aceticlastic methanogens were investigated. Completely mixed thermophilic anaerobic reactors operated at a chemical oxygen demand (COD) loading rate of 4 g/lday and a solid retention time (SRT) of 7 days were subjected to TAN concentrations of 0.40, 1.20, 3.05, 4.92, and 5.77 g/l. The reactor operations presented a case of chronic inhibition and it was observed that TAN concentrations of 4.92 and 5.77 g/l caused a drop in methane production by as much as 39% and 64%, respectively with respect to control. Batch anaerobic toxicity assays (ATA) were also performed to evaluate the acute toxicity effects of TAN and pH on methanogenesis at thermophilic condition. Modeling based on the results of ATA indicated that aceticlastic methanogens acclimated to high concentrations of TAN were less sensitive to increase in TAN and could tolerate wider pH ranges. TAN concentration causing 100% inhibition occurred in the range of 8-13 g/l, depending on acclimation condition and system pH. PMID- 12892666 TI - Comparison of modified montmorillonite adsorbents. Part II: The effects of the type of raw clays and modification conditions on the adsorption performance. AB - This paper builds on the preceding researches to study the effects of the type of clays (montmorillonites K10, KSF) and modifying conditions on the structure and adsorption behavior of resulting clay adsorbents. The raw clays were modified by polymeric Al/Fe species, hexadecyl-trimethylammonium (HDTMA) surfactant and a complex of polymeric Al/Fe-HDTMA. X-ray diffraction spectra was applied to analyze the structure of the raw and modified clays. After modification, the basal spacing of the clays varied, depending on the types of raw clay and modification conditions. Copper and phenol were selected as adsorbates for evaluating the adsorption performance of various clays, which was affected significantly by the types of raw clay and modification conditions. In general the inorganic contaminant (e.g., Cu) tend to be adsorbed by the polymeric Al/Fe modified clay and the organic impurities (e.g., phenol) will be preferably captured by the surfactant modified clay; both due to the specific surface properties resulting from introducing the modifiers. The complex modified clays possessed the ability of adsorbing both inorganic and organic contaminants. In addition, the d 0 0 1 spacing of modified KSF was greater than that of K10; the adsorption performance with modified KSF was thus greater than that with the modified K10. Finally, the ratio of modifiers to the clay (metal:surfactant:clay) has been observed to affect the adsorption performance; the optimal conditions have been defined. PMID- 12892667 TI - Studies on photo-degradation of 2,4-dinitro toluene in aqueous phase. AB - The efficiency of different photo-degradation processes was evaluated for degrading 2,4-dinitro toluene (DNT) in aqueous phase. The rate and extent of DNT degradation and removal of total organic carbon (TOC) and total nitrogen (TN) contents were compared for direct photolytic and photo-oxidative reactions using various concentrations of H2O2 and Fenton's reagent with a 125 W medium pressure UV lamp. DNT was degraded rapidly under photo-oxidative conditions. Complete destruction was obtained using Fenton's reagent, wherein 100 ppm of DNT was degraded within 60 min of irradiation time. Removal of TOC and TN contents in the photo-Fenton system was 96% and 57%, respectively, after 2 h of UV irradiation. Degradation of DNT followed first order reaction kinetics. Photo-Fenton oxidation is found to be the most suitable technique to degrade DNT in aqueous phase. PMID- 12892668 TI - Disinfection of surfaces by photocatalytic oxidation with titanium dioxide and UVA light. AB - Particularly in microbiological laboratories and areas in intensive medical use, regular and thorough disinfection of surfaces is required in order to reduce the numbers of bacteria and to prevent bacterial transmission. The conventional methods of disinfection with wiping are not effective in the longer term, cannot be standardized, are time- and staff-intensive and use aggressive chemicals. Disinfection with hard ultraviolet C (UVC) light is usually not satisfactory, as the depth of penetration is inadequate and there are occupational medicine risks. Photocatalytic oxidation on surfaces coated with titanium dioxide (TiO2) might offer a possible alternative. In the presence of water and oxygen, highly reactive OH-radicals are generated by TiO2 and mild ultraviolet A (UVA). These radicals are able to destroy bacteria, and may therefore be effective in reducing bacterial contamination. Direct irradiation with UVC however can produce areas of shadow in which bacteria are not inactivated. Using targeted light guidance and a light-guiding sheet (out of a UVA-transmittant, Plexiglas, for example), as in the method described in the present study, bacterial inactivation over the entire area is possible. The effectiveness of the method was demonstrated using bacteria relevant to hygiene such as Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus and Enterococcus faecium. For these bacteria, a reduction efficiency (RE) more than 6log10 steps in 60 min was observed. Using Candida albicans, a RE of 2log10 steps in 60 min was seen. Light and scanning electron microscopic examinations suggest that the germ destruction achieved takes place through direct damage to cell walls caused by OH-radicals. PMID- 12892669 TI - Ozonisation coupled with biological degradation for treatment of phenolic pollutants: a mechanistically based study. AB - Phenolic acids constitute an important group of pollutants which are reluctant to biological treatment. Solutions containing a mixture of cinnamic acid, p-coumaric acid, caffeic acid and ferulic acid were submitted to ozonisation. Then, the changes in biodegradability along the process were studied by means of respirometry. There is an optimum ozone dosage in the interval 3-5 min of treatment which allows to achieve the maximum increase in biodegradability (more than 10 times) and a high efficiency of the ozonisation process (COD decreases to a half of its initial value). Further ozonisation does not help to increase biodegradability and is clearly disadvantageous. Similar results are obtained with actual samples of olive oil wastewaters. This behaviour is explained based on the formation of highly biodegradable benzaldehydes as key ozonisation intermediates, in the early reaction stages. PMID- 12892670 TI - The influence of FeCl3 on the photocatalytic degradation of dissolved azo dyes in aqueous TiO2 suspensions. AB - The study concerned decolouration of solutions of azo, anionic (Acid Orange 7, Reactive Red 45, Acid Yellow 23) and cationic (Basic Blue 41 and Basic Orange 66) dyes during illumination with UV (lambdamax 366 nm) irradiation in the presence of TiO2 and FeCl3. The process of decolouration during illumination of the solutions studied containing FeCl3 underwent significant intensification in the case of anionic dyes and unfavourable inhibition in case of cationic dyes. It was also observed that FeCl3 had a diverse influence on the adsorption of the dyes studied on TiO2. The adsorption of anionic dyes and decolouration of solutions before the illumination was observed only in the presence of FeCl3. In case of cationic dyes the addition of FeCl3 caused elimination of these phenomena. An additional cause of decolouration of anionic dyes solutions before illumination was the precipitation of their poorly soluble compounds from Fe3+. The processes of degradation and mineralization of the dye that accompanied decolouration of Acid Orange 7 solutions were also observed. It was stated that similarly to the case of Acid Orange 7, the decolouration of the studied anionic dyes' solutions can depend on the concentration of FeCl3, the amount of TiO2 and the initial concentration of the dye in its solution. PMID- 12892671 TI - Improvement of thermal hydrolysis rate of dichloroacetic acid using alcohol. AB - Dichloroacetic acid (DCAA) is produced during the oxidation of trichloroethylene. It is also produced in drinking water treatment as a disinfection by-product. DCAA is a problem material, because of its toxicity. The objective of this research is to find the final products and the reaction pathway of the DCAA decomposition by hydrolysis, and to increase the hydrolysis rate. The removal of both chlorine atoms in DCAA structure was achieved with hydrolysis at around 75 degrees C, and the final products were oxalic acid and glycolic acid. The reaction pathway was the production of oxalic acid and glycolic acid from two glyoxylic acid molecules by Cannizzaro reaction after the glyoxylic acid production from dechlorination of DCAA with hydrolysis. The hydrolysis rate of DCAA was increased with the use of 90% ethanol solution as solvent. The activation energy of DCAA was about 80 kJ/mol in it, while it was around 105 kJ/mol in water. PMID- 12892672 TI - Anaerobic biodegradation of weathered polychlorinated biphenyls (PCBs) in contaminated sediments of Porto Marghera (Venice Lagoon, Italy). AB - The biodegradation of weathered polychlorinated biphenyls (PCBs) (mono and di chlorinated biphenyls along with PCBs partially ascribed to Aroclor 1242 and 1254) occurring at 1.5-2.5 mg/kg in three different sediments collected from the Porto Marghera contaminated area of Venice Lagoon (Italy) was reported in this study. Strictly anaerobic, slurry microcosms consisting of sediments suspended (at 25% v/v) in a marine salt medium, lagoon water or lagoon water supplemented with NaHCO3 and Na2S were developed and monitored for PCB transformation, sulfate consumption and methane (CH4) production for 6 months. A marked depletion of highly chlorinated biphenyls along with the accumulation of low-chlorinated, often ortho-substituted biphenyls was observed in the biologically active microcosms, where a remarkable consumption of sulfate and/or a significant production of CH4 were also detected. Notably, a more extensive PCB transformation was observed in the microcosms developed with site water (both without or with NaHCO3 plus Na2S), where both the initial concentration of sulfate and sulfate consumption were five fold-higher than in the corresponding microcosms with salt medium. These data indicate that weathered PCBs of the three contaminated sediments of Porto Marghera utilized in this study can undergo reductive dechlorination, probably mediated by indigenous sulfate-reducing and/or methanogenic bacteria. PMID- 12892673 TI - Plant uptake and translocation of air-borne chlordane and comparison with the soil-to-plant route. AB - In order to assess fully the impact of persistent organic pollutants (POPs) on human health, pollutant exchange at the interface between terrestrial plants, in particular food crops, and other environmental compartments must be thoroughly understood. In this regard, transfers of multicomponent and chiral pollutants are particularly informative. In the present study, zucchini (Cucurbita pepo L.) was planted in containerized, uncontaminated soil under both greenhouse and field conditions and exposed to air-borne chlordane contamination at 14.0 and 0.20 ng/m(3) (average, greenhouses), and 2.2 ng/m(3) (average, field). Chiral gas chromatography interfaced to an ion trap mass spectrometer was used to determine the chiral (trans-chlordane, TC, and cis-chlordane, CC) and achiral (trans nonachlor, TN) chlordane components in vegetation, air, and soil compartments. The chlordane components of interest were detected in all vegetation tissues examined--root, stem, leaves, and fruits. When compared with the data from a soil to-plant uptake study, the compositional profile of the chlordane components, i.e. the component fractions of TC, CC, and TN, in plant tissues, showed significantly different patterns between the air-to-plant and soil-to-plant pathways. Changes in the enantiomer fractions of TC and CC in plant tissues relative to the source, i.e. air or soil, although observed, were not markedly different between the two routes. This report provides the first comprehensive comparison between two distinct plant uptake routes for POPs and their subsequent translocation within plant tissues. PMID- 12892674 TI - Polycyclic aromatic hydrocarbons in weathered bottom ash from incineration of municipal solid waste. AB - Polycyclic aromatic hydrocarbons (PAHs) were analysed in weathered bottom ash from different municipal solid waste incinerators. The sum of the 16 "USEPA" PAHs was found to vary from 480 to 3590 microg/kg. The amounts of carcinogenic PAHs were between 89 and 438 microg/kg ash. The maximum levels of carcinogenic PAHs exceed the Swedish generic guidelines for sensitive land use. The results showed that the amount of PAHs in fresh and weathered bottom ash are similar, indicating that the PAHs are strongly bound to the ash and only limited amounts are released to the surroundings. PMID- 12892675 TI - Temporal and local trends of PCDD/F levels in cow's milk in Switzerland. AB - Levels of polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/F) were determined in 30 Swiss cow's milk samples collected at dairy farms in the vicinity to point sources, in rural/alpine areas distant to known sources, and from tanks in large industrial milk processing plants. The contaminant concentrations in samples collected in 2001 were compared to data from analyses conducted in 1984 and 1990/1991 at the same sites. In 2001, the PCDD/F levels in milk from farms near point sources (0.63+/-0.26 ng I-TEQ/kg milk fat) are slightly but significantly higher in than milk from remote areas (0.36+/-0.09 ng I-TEQ/kg milk fat). Consumer milk collected at the processing plants had intermediary levels (0.51+/-0.19 ng I-TEQ/kg milk fat). However, milk in 2001 was significantly less contaminated than the samples collected in 1990/1991 and 1984. This trend is particularly pronounced near point sources but is also apparent in consumer milk and milk from remote areas. No geographical gradient in the atmospheric input of PCDD/F in Switzerland was found. The reduction in PCDD/F levels in dairy milk is paralleled by and correlated to the remediation of known PCDD/F emitting industries, as enforced by federal authorities. PMID- 12892676 TI - Extraction and cleanup methods of dioxins in house dust from two cities in Japan using accelerated solvent extraction and a disposable multi-layer silica-gel cartridge. AB - A simple and rapid method for the extraction and cleanup of dioxins from house dust was developed using an accelerated solvent extraction (ASE) method and a multi-layer silica-gel cartridge. It was found that the WHO-TEQ levels of dioxins extracted from the house dust obtained by both a conventional soxhlet extraction and the ASE were almost equal, when the data obtained by both methods were compared. As for the cleanup method, a multi-layer silica-gel cartridge yielded higher dioxin recoveries than the alkaline digestion method. The average values of the dioxins in house dust from Kumagaya city and Sendai city in Japan (Sendai city is bigger than Kumagaya city with respect to the population and industry), were 15.6 pg TEQ/g (8.6-26.0 pg TEQ/g, n=5, Kumagaya city) and 16.0 pg TEQ/g (5.9 30.5 pg TEQ/g, n=5, Sendai city), respectively. PMID- 12892677 TI - The PCB pollution of Lake Iseo (N. Italy) and the role of biomagnification in the pelagic food web. AB - Several models of varying complexity have been used to predict pollutant concentrations in the higher levels of the food web from those in lower levels, but the role of the biomagnification process in aquatic food chains is still controversial. We used the fugacity-based approach to verify the transfer of PCBs through the pelagic food chain of Lake Iseo (N. Italy), sampling several zebra mussel specimens and some fish belonging of different trophic levels. The zebra mussel seems to be a suitable starting species for modelling the bioaccumulation process through the trophic web, not only because its physiological characteristics and population size do not change much with time (as do algae and zooplankton) but also because it takes up toxicants exclusively from the water, as shown by the application of two predictive trophic models commonly used. The data provided by one of those models were in good agreement with our experimental data on fish in Lake Iseo, that show a not negligible uptake from food for the top predator species (pike and perch) with an increase of about three times in comparison with the PCB levels measured in the zebra mussel specimens. PMID- 12892678 TI - The behavior of PCDD/Fs, PCBs, chlorobenzenes and chlorophenols in wet scrubbing system of municipal solid waste incinerator. AB - Though some researchers have already reported polychlorinated dibenzo-p-dioxins and polychlorinated dibenzofurans (PCDD/Fs) concentration in flue gas at inlet and outlet of wet scrubber (WS) in municipal solid waste incinerator (MSWI), there are few investigations about scrubbing water, suspended solid (SS) in scrubbing water and sludge. In order to understand the behavior of PCDD/Fs in WS and to make clear the effect of the residence time of scrubbing water in WS on the PCDD/Fs concentrations in scrubbing water and at the outlet gas of WS, PCDD/Fs, polychlorinated biphenyls (PCBs), chlorobenzenes (CBzs) and chlorophenols (CPs) concentrations in various media around WS were investigated at two MSWIs. PCDD/Fs concentration in flue gas at outlet of WS was higher than that at inlet of WS in both MSWIs. In WS A, it was observed that the homologue profiles changed considerably and the concentration of higher chlorinated compounds such as H7CDD/Fs and O8CDD/F increased remarkably at outlet gas of WS A. Only CPs were transferred from flue gas to scrubbing water in both WSs. As for scrubbing water, a large amount of their chlorinated aromatic compounds was detected in SS particles. According to the results, it was found that not only flue gas conditions but also operational and structural conditions in WS had great influence on minimizing the memory of chlorinated aromatic compounds in WS. PMID- 12892679 TI - Study on PCBs, PCDD/Fs, organochlorine pesticides, heavy metals and arsenic content in freshwater fish species from the River Turia (Spain). AB - In this study samples of common trout, European eel and barbel have been analysed for the levels of heavy metals such as lead (Pb), cadmium (Cd), zinc (Zn), copper (Cu), and the metalloid arsenic (As). Polychlorinated biphenyls (PCBs), including three non-ortho substituted PCBs, 2,3,7,8-substituted polychlorinated dibenzo-p dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs) and DDT and its two main metabolites were also analysed in some selected samples, in order to evaluate the extent of contamination of the River Turia basin. All samples presented detectable levels of the elements analysed, presenting zinc the highest concentrations. European eel was the species found to highly accumulate these contaminants. Regarding organochlorinated compounds, seven samples exhibited detectable levels of the most toxic PCDD/Fs congener, the 2,3,7,8-TCDD. Non-ortho PCBs were not detected or not quantified in almost all samples and the sum of the individual PCB congeners determined were low compared to other aquatic ecosystems with similar fish species. PMID- 12892680 TI - Chlorinated hydrocarbon contaminants in blood of black and turkey vultures from Savannah River Site, South Carolina, USA. AB - Blood of adult and juvenile black and turkey vultures in the Savannah River Site of South Carolina, USA was analyzed for the presence of 2,3,7,8-chlorine substituted polychlorinated dibenzo-p-dioxins (2,3,7,8-PCDDs), dibenzofurans (2,3,7,8-PCDFs), -dioxin-like and -di-ortho polychlorinated biphenyls. Concentration ranges of 2,3,7,8-PCDD/DFs in blood of black and turkey vultures were 14.2-34.6 and 8.7-56.2 pg/ml wet wt., respectively. Dioxin-like PCBs were in the ranges of 815-4627 and 753-3611 pg/ml wet wt. respectively, in black and turkey vultures. Considerable concentrations of two congeners of di-ortho PCBs were noticed in the ranges of 1415-10325 and 663-7500 pg/ml respectively, in black and turkey vultures. Comparatively, greater toxic equivalency (TEQ) were observed in blood of turkey vultures with the ranges of 3.2-20, whereas black vulture contained 1.8-8.4 pgTEQ/ml wet wt. basis. The species-specific accumulation profiles of PCDD/DFs and dioxin-like PCBs may reflect the different feeding habits, ecology, metabolic capacity and migratory movements of these two scavenging species. PMID- 12892681 TI - Six interaction profiles for simple mixtures. AB - The Agency for Toxic Substances and Disease Registry (ATSDR) has a program for chemical mixtures that encompasses research on chemical mixtures toxicity, health risk assessment, and development of innovative computational methods. ATSDR prepared a guidance document that instructs users on how to conduct health risk assessment on chemical mixtures (Guidance Manual for the Assessment of Joint Toxic Action of Chemical Mixtures). ATSDR also developed six interaction profiles for chemical mixtures. Two profiles were developed for persistent environmental chemicals that are often found in contaminated fish and also can be detected in human breast milk. The mixture included chlorinated dibenzo-p-dioxins, hexachlorobenzene, dichlorodiphenyl dichloroethane, methyl mercury, and polychlorinated biphenyls. Two profiles each were developed for mixtures of metals and mixtures of volatile organic chemicals (VOCs) that are frequently found at hazardous waste sites. The two metal profiles dealt with (a) lead, manganese, zinc, and copper; and (b) arsenic, cadmium, chromium, and lead; the two VOCs mixtures dealt with (a) 1,1,1-trichloroethane, 1,1-dichloroethane, trichloroethylene, and tetrachloroethylene; and (b) benzene, ethylbenzene, toluene, and xylenes (BTEX). Weight-of-evidence methodology was used to assess the joint toxic action for most of the mixtures. Physiologically based pharmacokinetic modeling was used for BTEX. In most cases, a target-organ toxicity dose modification of the hazard index approach is recommended for conducting exposure-based assessments of noncancer health hazards. PMID- 12892682 TI - Costing the cascade: estimating the cost of increased obstetric intervention in childbirth using population data. AB - OBJECTIVE: To estimate the cost of "the cascade" of obstetric interventions introduced during labour for low risk women. DESIGN: A cost formula derived from population data. SETTING: New South Wales, Australia. POPULATION: All 171,157 women having a live baby during 1996 and 1997. METHODS: Four groups of interventions that occur during labour were identified. A cost model was constructed using the known age-adjusted rates for low risk women having one of three birth outcomes following these pre-specified interventions. Costs were based on statewide averages for the cost of labour and birth in hospital. MAIN OUTCOME MEASURES: The outcome measure is an "average cost unit per woman" for low risk women, predicted by the level of intervention during labour. Obstetric care is classified as either private obstetric care in a private or public hospital, or routine public hospital care. RESULTS: The relative cost of birth increased by up to 50% for low risk primiparous women and up to 36% for low risk multiparous women as labour interventions accumulated. An epidural was associated with a sharp increase in cost of up to 32% for some primiparous low risk women, and up to 36% for some multiparous low risk women. Private obstetric care increased the overall relative cost by 9% for primiparous low risk women and 4% for multiparous low risk women. CONCLUSIONS: The initiation of a cascade of obstetric interventions during labour for low risk women is costly to the health system. Private obstetric care adds further to the cost of care for low risk women. PMID- 12892683 TI - Levels of antibodies against cytomegalovirus and Chlamydophila pneumoniae are increased in early onset pre-eclampsia. AB - OBJECTIVE: The origins of pre-eclampsia/eclampsia lie in a mismatch between feto placental demands and utero-placental supply, a situation that also arises in normotensive intrauterine growth restriction (IUGR). Could reactivated chronic infection be both a trigger for these differential maternal responses to the same underlying pathology and a link between pre-eclampsia and its attendant lifelong risks of atherosclerosis? DESIGN: Nested case-control study. SETTING: Tertiary obstetric centre. POPULATION: Cases of pre-eclampsia, normotensive IUGR and controls. METHODS: A nested case-control study of serum from a population-based bank was performed. Seroprevalence and levels of anti-cytomegalovirus (CMV) and Chlamydophila pneumoniae immunoglobulin G (IgG) were compared (non parametrically) between women with early onset pre-eclampsia (<34 weeks of gestation, n = 9), late onset pre-eclampsia (> or =34 + 0 weeks of gestation, n = 29), normotensive IUGR (birthweight less than third centile, n = 33) and matched normal pregnancy (n = 113, up to 2 per case). RESULTS: There was a significant difference in both anti-CMV and Chl. pneumoniae antibodies between groups (Kruskal-Wallis test, P < 0.05). Women with early onset pre-eclampsia had higher anti-CMV levels (median: 79, 95% confidence interval [95% CI] = 47, 164) than women with late onset pre-eclampsia (26 [95% CI = 22, 82], P < 0.05), normotensive IUGR (40 [95% CI = 31, 72], P < 0.05) and normal pregnancy (49 [95% CI = 45, 70], P < 0.05). Women with normotensive IUGR had significantly lower anti-Chl. pneumoniae antibodies (0.10 [95% CI = 0.08, 0.38]) than did normal pregnancy controls (0.21 [95% CI = 0.20, 0.28], P <0.05). CONCLUSIONS: The anti CMV and anti-Chl. pneumoniae antibodies were higher in early onset pre-eclampsia than in late onset pre-eclampsia, normotensive IUGR and normal pregnancy. This may provide a pathophysiological link between pre-eclampsia and the known increased risk for subsequent atherosclerosis. PMID- 12892684 TI - The in vitro effect of dual combinations of ritodrine, nicardipine and atosiban on contractility of pregnant rat myometrium. AB - OBJECTIVE: To compare the tocolytic potency of ritodrine, nicardipine and atosiban, used alone and in dual combinations, to see whether combinations of these drugs, which act via different pathways, could improve inhibition of uterine contractility. DESIGN: Study on myometrial contractility in vitro. SETTING: Laboratory of physiology, Lyon, France. SAMPLE: Longitudinal myometrial strips from non-labouring timed pregnant Wistar rats (18 gestational days). METHODS: Strips were simultaneously exposed to EC(25), EC(50) or EC(75) of dual combinations of either ritodrine and nicardipine, ritodrine and atosiban or nicardipine and atosiban (n = 10/group). Basal contractile activity and contractile activity after addition of each combination was measured using the 10 min integral of activity. Changes were expressed as percentage from the basal 10 min integral activity. The observed percentage inhibition of activity was compared with the expected percentage inhibition in an additive pharmacological model. When no significant difference occurred, the combination was deemed simply to have an additive tocolytic effect. When inhibition of activity was significantly greater compared with the expected percentage inhibition, the combination was deemed to have a synergistic effect. MAIN OUTCOME MEASURE: Changes in integral contractile activity in response to tocolytic combinations. RESULTS: Ritodrine and atosiban inhibited integral activity to a greater extent than expected [e.g. using EC(50): observed inhibition 88.9% (13.8%) vs expected inhibition 75%; P < 0.015]. Actual inhibition by nicardipine/ritodrine [78.55% (20.4%) vs 75%; P = n.s.] and nicardipine/atosiban [78.94% (17.8%) vs 75%; P = n.s.] was not significantly different from expected. CONCLUSIONS: A combination of ritodrine plus atosiban exhibits a synergistic inhibition for myometrial activity, thus allowing the use of lower concentrations of each drug to achieve the same effect compared with each drug used alone. Combination of nicardipine plus ritodrine and nicardipine plus atosiban achieves only an additive effect. The potential for decreasing side effects (beta-mimetics) and costs (atosiban) when using a combination in clinical practice needs to be evaluated. PMID- 12892685 TI - The effect of systemic administration of lipopolysaccharide on cerebral haemodynamics and oxygenation in the 0.65 gestation ovine fetus in utero. AB - OBJECTIVE: To investigate the effect of intravenous lipopolysaccharide on systemic and cerebral haemodynamics and oxygenation in the preterm ovine fetus. DESIGN: Prospective observational study. SETTING: Research centre for perinatal brain injury. SAMPLE: Nine fetal sheep at circa 93 days of gestation (0.65). METHODS: Fetal sheep were chronically instrumented with arterial and venous catheters and a flow probe in the carotid artery. Near-infrared spectroscopy was used to measure changes in cerebral oxygenation and total haemoglobin concentration. Three days after surgery, each fetus was given 100 ng/kg Escherichia coli lipopolysaccharide. Observations were continued for 48 hours post-injection and compared with baseline control values. MAIN OUTCOME MEASURES: Fetal heart rate, mean arterial pressure, carotid blood flow. RESULTS: Three fetuses died after administration of the lipopolysaccharide. In the survivors fetal heart rate rose from 193 (SEM 7) to a mean maximal level of 226 (SEM 31 bpm) (P = 0.01) after 6.5 (SEM 1.0) hours. The mean arterial pressure decreased from 40.5 (SEM 4.2) to 29.4 (SEM 1.6) mmHg (P < 0.05) after 7.0 (SEM 2.0) hours, and carotid blood flow increased from 29.6 (SEM 1.6) to 45.8 (SEM 5.7) mL/min (P = 0.0002) at 12 (SEM 3) hours. All values returned to control levels by 48 hours. Histological assessment showed evidence of periventricular leucomalacia in three out of six brains studied. CONCLUSION: These data do not suggest that cerebral ischaemia is the main aetiological factor in endotoxin-related fetal brain injury. Fetal tachycardia and cerebral vasodilation may indicate endotoxaemia in fetuses exposed to prenatal infection. PMID- 12892686 TI - Midwives' perceptions and experiences of routine enquiry for domestic violence. AB - OBJECTIVE: To examine midwives' perceptions and experiences of routine enquiry for domestic violence. DESIGN: Focus groups and semi-structured interviews. SETTING: The study was conducted in the maternity services of Guy's and St Thomas' NHS Hospital Trust, an inner London teaching hospital. SAMPLE: Twenty eight midwives were interviewed. METHODS: One hundred and sixteen out of 145 (80%) midwives were trained to screen routinely pregnant women for domestic violence at booking, at 34 weeks of gestation and postpartum. Twenty midwives attended one of four focus groups and eight semi-structured interviews were conducted. Interviews were transcribed and analysed using content analysis. MAIN OUTCOME MEASURES: Midwives' views of routine enquiry for domestic violence. RESULTS: Midwives felt that domestic violence was an important issue to be addressed. However, practical and personal difficulties were encountered during the study. These included: time constraints, lack of confidential time, safety issues, staff shortages, low staff morale and midwives' personal experiences of domestic violence. The study produced a raised awareness about domestic violence within the maternity unit. There was considerable support for the idea of a specialist domestic violence midwife. CONCLUSIONS: Routine enquiry for domestic violence cannot be implemented effectively without ensuring that in-depth training, resources, staff support and policies to ensure that screening can be conducted safely and confidentially are in place. PMID- 12892687 TI - The effect of a novel vaginal ring delivering oestradiol acetate on climacteric symptoms in postmenopausal women. AB - OBJECTIVE: Evaluate the efficacy and acceptability of a novel vaginal ring delivering oestradiol acetate (Menoring) versus oral oestradiol for relief of climacteric symptoms in postmenopausal women. DESIGN: Prospective, multicentre, randomised, double-blind, comparator-controlled, parallel group study. SETTING: Twenty-one centres in the United Kingdom. POPULATION: Postmenopausal women, aged <65 years experiencing > or =20 hot flushes/night sweats per week for two consecutive weeks. METHODS: Patients received a vaginal ring releasing oestradiol acetate at a rate equivalent to 50 microg/day in addition to placebo tablets (vaginal ring group) or oral oestradiol 1 mg/day in addition to placebo vaginal ring (oral group) for 24 weeks. For patients with inadequate control of vasomotor symptoms, dosage was doubled at 12 weeks. Non-hysterectomised women received norethisterone 1 mg/day for 12 days in each 28 day cycle. MAIN OUTCOME MEASURES: Efficacy was assessed by change in climacteric symptoms measured by the Greene Climacteric Scale, as well as mean change in frequency of hot flushes/night sweats. RESULTS: A total of 159 patients were enrolled (84 vaginal ring, 75 oral). Significant improvement (P < 0.05) in mean total Greene Climacteric Scale scores in both treatment groups was observed at 12 and 24 weeks and for every subscale score including anxiety, depression and sexual dysfunction (P < 0.05). In both groups, frequency of hot flushes/night sweats was significantly reduced (P < 0.001) at 12 and 24 weeks. No significant between-group differences were noted at 12 or 24 weeks. CONCLUSIONS: The oestradiol vaginal ring significantly improved climacteric symptoms as measured by Greene Climacteric Scale scores and reduced the frequency of hot flushes/night sweats. Efficacy and safety of vaginal ring oestradiol were comparable with those of oral therapy. Patient evaluations of oestradiol vaginal ring tolerability and acceptability were excellent. PMID- 12892688 TI - LH levels in women with polycystic ovarian syndrome: have modern assays made them irrelevant? AB - OBJECTIVE: To determine whether using newer monoclonal rather than polyclonal assays for measuring luteinising hormone (LH) alters the predictive value of LH and LH/follicle-stimulating hormone (FSH) ratios for polycystic ovarian syndrome. DESIGN: Prospective cohort study. SETTING: Fertility and Reproductive Endocrinology Clinic within a New Zealand Teaching hospital. POPULATION: Seventy eight women presenting with oligomenorrhoea or hirsutism and polycystic ovaries on pelvic ultrasound and 59 volunteer controls with ultrasonically normal ovaries and a regular menstrual cycle. METHODS: Serum LH concentrations were measured using a polyclonal radio-immunoassay (Amerlex-M, Johnson & Johnson) and two monoclonal immunometric assays (Immulite, DPC; Cobas Core, Hoffman La Roche). The proportion of women with an elevated serum LH concentration in each group was calculated using both current local laboratory reference intervals and a new reference range derived from our control group. The LH/FSH ratios for women in both groups were also calculated using the three different LH assays. MAIN OUTCOME MEASURES: LH concentrations and LH:FSH ratios measured using polyclonal and monoclonal immunoassays. RESULTS: Using the local laboratory normal range, a significantly higher proportion of women had an elevated LH when measured with a polyclonal assay (23.1%) than when measured with a monoclonal assay (12.8% Core, 6.4% Immulite) (P < 0.05). LH/FSH ratios were significantly lower when monoclonal assay was used and receiver-operator characteristic curves suggest that LH/FSH ratios of 1 or lower provide the most reliable separation of women with polycystic ovarian syndrome from controls when these assays are used. CONCLUSIONS: Clinicians should be aware that the use of monoclonal LH assays will result in significantly lower measured LH levels and LH/FSH ratios in women with polycystic ovarian syndrome than previously used polyclonal assays. Account should be taken of the assay type used, when using endocrinological parameters in the diagnosis of polycystic ovarian syndrome, or the identification of women who have LH hypersecretion. PMID- 12892689 TI - Improved fertility following conservative surgical treatment of ectopic pregnancy. AB - OBJECTIVE: To evaluate fertility after salpingectomy or tubotomy for ectopic pregnancy. DESIGN: Retrospective cohort study. SETTING: Clinical University Center, Hvidovre Hospital, Copenhagen. POPULATION: Two hundred and seventy-six women undergoing salpingectomy or tubotomy for their first ectopic pregnancy between January 1992 and January 1999 and who actively attempted to conceive were followed for a minimum of 18 months. METHODS: Retrospective cohort study combined with questionnaire to compare reproductive outcome following salpingectomy or tubotomy for ectopic pregnancy. Cumulative probabilities of pregnancy for each group were calculated by the Kaplan-Meier estimator and compared by Cox regression analysis to control for potential confounders. MAIN OUTCOME MEASURES: Intrauterine pregnancy rates and recurrence rates of ectopic pregnancy after surgery for ectopic pregnancy. RESULTS: The cumulative intrauterine pregnancy rate was significantly higher after tubotomy (88%) than after salpingectomy (66%) (log rank P < 0.05) after correction for confounding factors. No difference was found in the recurrence rate of ectopic pregnancy between the treatments (16% vs 17%). In patients with contralateral tubal pathology, the chance of pregnancy was poor (hazard ratio 0.463) and the risk of recurrence was high (hazard ratio 2.25), assessed with Cox regression. The rate of persistent ectopic pregnancy was 8%. CONCLUSION: Conservative surgery is superior to radical surgery at preserving fertility. Conservative surgery is not followed by an increased risk of repeat ectopic pregnancy, but by the risk of persistent ectopic pregnancy, which should be taken into account when deciding on the operative procedure. Management in case of contralateral tubal pathology is disputed and should ideally be addressed in a randomised clinical trial. PMID- 12892690 TI - The influence of pad test loss on management of women with urodynamic stress incontinence. AB - Members of the International Continence Society (UK section) were randomised to receive a scenario comprising clinical and urodynamic data of a woman with urodynamic stress incontinence, including a one-hour pad loss of either 42 g (large loss) or 7 g (small loss). Members were asked to indicate their initial management choice from a list of four options. Three hundred and fifteen of 440 (72%) members responded. Gynaecologists and urologists were significantly more likely to choose surgical treatment for patients with a large pad loss (24% vs 8%; difference 15.8%, 95% CI 2.9-28.7), despite published data demonstrating the poor reproducibility and reliability of the pad test. PMID- 12892691 TI - Reproducibility and reliability of pressure flow parameters in women. AB - OBJECTIVE: The aim of this study was to evaluate the intra- and inter-rater reliability of pressure flow parameters in women. DESIGN: A prospective study. SETTING: Tertiary referral unit in a London teaching hospital. POPULATION: Urodynamic traces of 621 women were studied to assess the reproducibility and reliability of pressure flow parameters. METHODS: Women with lower urinary tract symptoms were studied prospectively. Opening detrusor pressure (ODP), detrusor pressure at maximum flow rate P(detQ(max)), maximum flow rate (Q(max)) and closure detrusor pressure (CDP) were measured by the same observer twice, one week apart. Pressure flows were compared using different media for filling (saline and contrast). All parameters were also measured in nine women from multiple voids during ambulatory urodynamics to assess their reproducibility and finally were remeasured by a second observer. RESULTS: There was good intra observer agreement for all pressure flow parameters except detrusor pressure at maximum flow rate and good inter-observer agreement for opening detrusor pressure, closure detrusor pressure and detrusor pressure at maximum flow rate. There was no significant difference between parameters using saline or contrast (P > 0.05, independent samples t test). CONCLUSION: Pressure flow parameters appear to be reproducible and consistent urodynamic measurements in women. PMID- 12892692 TI - Interaction between hormone replacement therapy preparations and oral anticoagulant therapy. AB - There is limited information regarding the interaction of hormone replacement therapy (HRT) and oral anticoagulants. Following the acute over-anticoagulation of two women shortly after the initiation of tibolone, we have undertaken a retrospective review of anticoagulated women to determine if the dose of oral anticoagulants and quality of anticoagulant control are affected by the introduction of HRT. We demonstrate that acute over-anticoagulation consistently occurs following the commencement of tibolone and requires anticoagulant dose modification to re-establish target international normalised ratio (INR). In contrast, non-tibolone HRT preparations do not consistently alter anticoagulant control or dose requirements. PMID- 12892693 TI - Occupational risk of Pasteurella multocida septicaemia and premature labour in a pregnant vet. PMID- 12892694 TI - A case of failed fibroid embolisation due to an unusual vascular supply. PMID- 12892695 TI - The unfacts of "request" caesarean section. PMID- 12892696 TI - Secondary colposuspension: results of a prospective study from a tertiary centre. PMID- 12892697 TI - Accuracy of outpatient biopsy in the diagnosis of endometrial cancer: a systematic quantitative review. PMID- 12892698 TI - Posterior fossa haemorrhage in a preterm infant following vacuum assisted delivery. PMID- 12892699 TI - The value of symphysiotomy. PMID- 12892700 TI - Can pre-operative computed tomography predict resectability of ovarian carcinoma at primary laparotomy? PMID- 12892701 TI - Changes in vaginal flora after 2% clindamycin vaginal cream in women at high risk of preterm birth. PMID- 12892702 TI - Listening to patients with unexplained menstrual symptoms: what do they tell the gynaecologist? PMID- 12892703 TI - Successful microwave ablation of endometrial carcinoma. PMID- 12892705 TI - Slit-Robo: neuronal guides signal in tumor angiogenesis. AB - Slit and Roundabout (Robo) are well-characterized for neuron and leukocyte guidance. Their governing roles have now been expanded to control tumor endothelial cell communication and mediate tumor-induced angiogenesis. PMID- 12892706 TI - A bad rap: Rap1 signaling and oncogenesis. AB - In the paper by Ishida et al. in this issue of Cancer Cell, the authors report the results of targeted inactivation of a Rap1-specific GTPase-activating protein (GAP) gene, called SPA-1, in mice. Rap1 hyperactivation was observed in hematopoietic cells, which led over time to features associated with symptoms typical of human myeloid dyslastic and myeloid proliferative diseases. The authors present additional data showing that the level of Rap1 activation is important for regulating myelopoiesis and that, in the right context, can deliver an oncogenic signal. PMID- 12892707 TI - Evidence emerges for early metastasis and parallel evolution of primary and metastatic tumors. AB - Tumor progression to metastasis usually is assumed to occur through clonal genomic and epigenetic evolution. However, present evidence that challenges this paradigm. They show that genomic aberrations in tumor cells disseminated in the bone marrows of patients with no clinical evidence of metastasis generally do not resemble the aberrations in the primary tumors from which they arose. They interpret this to mean that tumor cells disseminate very early and evolve to metastatic disease independent from the primary tumor. Their model suggests that adjuvant therapies should be targeted to lesions in the disseminated cells rather than lesions found in primary tumors. PMID- 12892708 TI - Getting oncolytic virus therapies off the ground. AB - An international meeting was held on the development and application of replicating viruses for cancer therapy this past March in Banff, Alberta. In this review, using the presentations at this meeting as a backdrop, we discuss how recent scientific and clinical findings are reshaping the development of oncolytic virus therapeutics. Here we identify some of the obstacles that these therapeutics face and discuss evolving strategies, both preclinically and clinically, that are facilitating oncolytic virus development. PMID- 12892709 TI - Histone deacetylase inhibitors in cancer therapy: is transcription the primary target? PMID- 12892710 TI - Induction of tumor angiogenesis by Slit-Robo signaling and inhibition of cancer growth by blocking Robo activity. AB - Slit is a secreted protein known to function through the Roundabout (Robo) receptor as a chemorepellent in axon guidance and neuronal migration, and as an inhibitor in leukocyte chemotaxis. Here we show Slit2 expression in a large number of solid tumors and Robo1 expression in vascular endothelial cells. Recombinant Slit2 protein attracted endothelial cells and promoted tube formation in a Robo1- and phosphatidylinositol kinase-dependent manner. Neutralization of Robo1 reduced the microvessel density and the tumor mass of human malignant melanoma A375 cells in vivo. These findings demonstrate the angiogenic function of Slit-Robo signaling, reveal a mechanism in mediating the crosstalk between cancer cells and endothelial cells, and indicate the effectiveness of blocking this signaling pathway in treating cancers. PMID- 12892711 TI - Selective inhibition of tumor microvascular permeability by cavtratin blocks tumor progression in mice. AB - Tumor vasculature is hyperpermeable to macromolecules compared to normal vasculature; however, the relationship between tumor hyperpermeability and tumor progression is poorly understood. Here we show that a cell-permeable peptide derived from caveolin-1, termed cavtratin, reduces microvascular hyperpermeability and delays tumor progression in mice. These antipermeability and antitumor actions of cavtratin occur in the absence of direct cytostatic or antiangiogenic effects. Cavtratin blocks microvascular permeability by inhibiting endothelial nitric oxide synthase (eNOS), as the antipermeability and antitumor actions of cavtratin are markedly diminished in eNOS knockout mice. Our results support the concepts that hyperpermeability of tumor blood vessels contributes to tumor progression and that blockade of eNOS may be exploited as a novel target for antitumor therapy. PMID- 12892712 TI - Identification of potential anticancer drug targets through the selection of growth-inhibitory genetic suppressor elements. AB - To identify human genes required for tumor cell growth, transcriptome-scale selection was used to isolate genetic suppressor elements (GSEs) inhibiting breast carcinoma cell growth. Growth-inhibitory GSEs (cDNA fragments that counteract their cognate gene) were selected from 57 genes, including known positive regulators of cell growth or carcinogenesis as well as genes that have not been previously implicated in cell proliferation. Many GSE-cognate genes encode transcription factors (such as STAT and AP-1) and signal transduction proteins. Monoclonal antibodies against a cell surface protein identified by GSE selection, neural cell adhesion molecule L1CAM, strongly inhibited the growth of several tumor cell lines but not of untransformed cells. Hence, selection for growth-inhibitory GSEs allows one to find potential targets for new anticancer drugs. PMID- 12892713 TI - Myeloproliferative stem cell disorders by deregulated Rap1 activation in SPA-1 deficient mice. AB - SPA-1 (signal-induced proliferation-associated gene-1) is a principal Rap1 GTPase activating protein in hematopoietic progenitors. SPA-1-deficient mice developed a spectrum of myeloid disorders that resembled human chronic myelogenous leukemia (CML) in chronic phase, CML in blast crisis, and myelodysplastic syndrome as well as anemia. Preleukemic SPA-1-deficient mice revealed selective expansion of marrow pluripotential hematopoietic progenitors, which showed abnormal Rap1GTP accumulation. Overexpression of an active form of Rap1 promoted the proliferation of normal hematopoietic progenitors, while SPA-1 overexpression markedly suppressed it. Furthermore, restoring SPA-1 gene in a SPA-1-deficient leukemic blast cell line resulted in the dissolution of Rap1GTP accumulation and concomitant loss of the leukemogenicity in vivo. These results unveiled a role of Rap1 in myeloproliferative stem cell disorders and a tumor suppressor function of SPA-1. PMID- 12892714 TI - ERK-MAPK signaling coordinately regulates activity of Rac1 and RhoA for tumor cell motility. AB - We describe two signaling events downstream of ERK-MAP kinase contributing to cell motility in colon carcinoma cells. The Fos family member Fra-1 is expressed in an ERK-dependent manner. Silencing of Fra-1 expression with short interfering RNAs leads to losses of cell polarization, motility, and invasiveness in vitro. These effects of ablating Fra-1 are a consequence of activation of a RhoA-ROCK pathway by beta1-integrin, leading to an increase in the amount of stress fibers and stabilization of focal adhesions. We propose that Fra-1 promotes cell motility by inactivating beta1-integrin and keeping RhoA activity low. This depression of RhoA activity is necessary to permit a second ERK-dependent signaling event via uPAR, the receptor for urokinase-type plasminogen activator, to activate Rac and to drive motility through polarized lamellipodia extension. PMID- 12892715 TI - Hurling-related hand injuries. AB - Hurling is a contact sport, associated with significant morbidity. We have identified specific hand injuries sustained by participants and quantified the functional and financial implications of these injuries. Over a 3-month period, all hand injuries seen in the fracture clinic of our regional trauma unit were studied prospectively. Of the 123 consecutive injuries, 41 (33%) were sustained during hurling matches. Metacarpal (47%) and proximal phalangeal (37%) fractures were the most frequent. Eight hurlers (20%) required surgical intervention. Only four (10%) of the injured players were wearing hand protection. The mean cost of injury to the player was pound 615. We suggest the introduction of the mandatory use of hand protection for hurling. PMID- 12892716 TI - Which tissue adhesive for wounds? AB - We studied the three available tissue adhesives comparing their ease of technique, wound healing, satisfaction, merits and complications when treating childhood lacerations. Children presenting with uncomplicated wounds <2.5cm and <6h since the injury were studied. There were 17 children in each group. Results were compared for the individual tissue adhesive and the technique-contact and non-contact. The application was considered pain free in 82% of the non-contact technique and 56% for the contact technique-pain in 18 and 44%, respectively (P=0.062). Parents were satisfied in 88 and 94% for the contact and non-contact techniques, respectively (P=0.505) and the authors in 76 and 94% (P=0.119). The glove stuck to the wound in nine instances and was damaged once while breaking the container. The scab persisted in all scalp applications for 9-25 days. The adhesive effect was similar in all three. Indermil was considered to be the best among the three. Non-contact, droplet instillation (rather than contact application as was suggested for Dermabond and Histoacryl) was felt more comfortable. PMID- 12892717 TI - Acute lateral dislocation of the patella: correlation of ultrasound scanning with operative findings. AB - Acute lateral dislocation of the patella has been associated with disruption of the medial restraints of the patella. Following non-operative management there is a re-dislocation rate of up to 44%. The purpose of this study was to test whether sonography is a reliable method of assessing the medial retinaculum after acute dislocation of the patella. Ten patients following acute patellar dislocation had an ultrasound scan (USS) performed by an experienced musculoskeletal radiologist. Each patient subsequently had an examination under anaesthetic, arthroscopy, and repair of the ruptured structures. The ultrasound reports were compared to the surgical findings to determine the accuracy of this investigation. USS located deficiencies in the ligamentous attachments to the medial border of the patella and the presence of avulsed bony fragments, all of which were confirmed at operation. The sonographic diagnosis of haematoma or torn fibres in the vastus medialis obliquus (VMO) corresponded with our operative findings. The most significant findings were the correlation of free fluid around the medial collateral ligament (MCL) with avulsion of the femoral attachment of the medial patellofemoral ligament (MPFL) and the presence of avulsed fragments of bone from the medial border of the patella. PMID- 12892722 TI - The density and protein content of calcium oxalate crystals precipitated from human urine: a tool to investigate ultrastructure and the fractional volume occupied by organic matrix. AB - One of the key debates in biomineralisation studies is the extent to which components of the organic matrix become occluded into the crystal lattice during growth. Here, the relationship between protein content and density of calcium oxalate crystals grown in human urine has been investigated in order to determine which fraction of crystal volume is non-mineral. The density of crystals varied from 1.84 to 2.08 g/cm3 while the protein content ranged from 0.1 to 2.1% (w/w). There was an inverse relationship between measured density and protein content which was qualitatively and quantitatively consistent with predictions based on reasonable densities for the mineral and non-mineral components. The coefficients of the fitted equation suggest that, at 2% protein (w/w), the volume of non mineral would be 5.0% (v/v). The density values we observed are incompatible with fractional volumes of 20%. The results confirm that the occlusion of a small but possibly significant amount of protein into a crystal lattice is possible, but cast doubt on the hypothesis that protein acts as a major intracrystalline ultrastructural element. Moreover, the methodology developed for this study offers a simple and robust method for interrogating organic/inorganic associations in a range of biological and medical systems. PMID- 12892721 TI - Anti-cementoblastoma-derived protein antibody partially inhibits mineralization on a cementoblastic cell line. AB - The effect of human anti-cementoblastoma-derived protein antibody during cementogenesis in vitro was investigated by using human cementoblastoma-derived cells. Cultures treated with 5 microg/ml of CP antibody from day 1 to day 15 revealed a significant decrease in alkaline phosphatase activity (ALP) 40% (p < 0.005), 44% (p < 0.001), 49% (p < 0.1), and 45% (p < 0.02) at 9, 11, 13, and 15 days, respectively. Immunoexpression of osteopontin revealed that in cultures treated with anti-CP antibody, the positive number of cementoblastoma cells was reduced by 87, 83, 69, and 52% at 5, 7, 9, and 11 days, respectively. Bone sialoprotein immunoexpression showed a decrease in positive cells of 82, 51, 60, 80, 83, and 87% at 5, 7, 9, 11, 13, and 15 days, respectively, as compared to controls. The Ca/P ratio of the mineral-like tissue deposited in vitro by cementoblastoma cells revealed that control cultures had a Ca/P ratio of 1.45 and 1.61 at 5 and 15 days, whereas experimental cultures revealed a Ca/P ratio of 0.50 and 0.79 at 5 and 15 days, respectively. Electron diffraction patterns showed inner double rings representing D-spacing that were consistent with those of hydroxyapatite in both control and experimental cultures. Examination of the crystallinity with high resolution transmission electron microscopy showed homogeneous and preferential spatial arrangement of hydroxyapatite crystallites in control and experimental cultures at 15 days. Atomic force microscopy images of control cultures at 5 and 15 days revealed small granular particles and grain agglomeration that favored the formation of crystalline plaques with a lamellar like pattern of the mineral-like tissue. Experimental cultures at 5 and 15 days showed tiny and homogeneous granular morphology. The agglomerates maintained spherical morphology without organization of needle-like crystals to form plaque like structures. Based on these findings, it is hypothesized that cementoblastoma derived protein may be associated to crystal growth, compositional and morphological features during the mineralization process of cementum in vitro. PMID- 12892723 TI - Focus gradient correction applied to tilt series image data used in electron tomography. AB - The resolution in 3D reconstructions from tilt series is limited to the information below the first zero of the contrast transfer function unless the signal is corrected computationally. The restoration is usually based on the assumption of a linear space-invariant system and a linear relationship between object mass density and observed image contrast. The space-invariant model is no longer valid when applied to tilted micrographs because the defocus varies in a direction perpendicular to the tilt axis and with it the shape of the associated point spread function. In this paper, a method is presented for determining the defocus gradient in thin specimens such as sections and 2D crystals, and for restoration of the images subsequently used for 3D reconstruction. The alignment procedure for 3D reconstruction includes area matching and tilt geometry refinement. A map with limited resolution computed from uncorrected micrographs is compared to a volume computed from corrected micrographs with extended resolution. PMID- 12892724 TI - Revisiting the structure of Alvinella pompejana hemoglobin at 20A resolution by cryoelectron microscopy. AB - The hemoglobin of the polychaete worm Alvinella pompejana was reconstructed at 20A resolution from frozen-hydrated samples observed by electron microscopy according to the random conical tilt series method. This three-dimensional reconstruction was mirror-inverted with respect to a previous volume published by de Haas et al. in 1996. In order to explain this handedness discrepancy, various 3D reconstructions using different reference volumes were carried out showing that the choice of the first volume was the keystone during the refinement process. The 3D reconstruction volume of A. pompejana Hb presented structural features characteristic of annelid Hbs with two hexagonal layers each comprising six hollow globular subassemblies and a complex of non-heme linker chains. Moreover, the eclipsed conformation of the two hexagonal layers and a HGS architecture similar to that described for Arenicola marina Hb led to the conclusion that A. pompejana Hb belonged to the architectural type II according to the definition of Jouan et al. (2001). A comparison between this cryo-EM volume and X-ray crystallography density maps of Lumbricus terrestris type-I Hb (Royer et al., 2000) showed that the triple stranded coiled coil structures of linker chains were different. Based on this observation, a model was proposed to explain the eclipsed conformation of the two hexagonal layers of type-II Hbs. PMID- 12892725 TI - The mechanical properties of simple epithelial keratins 8 and 18: discriminating between interfacial and bulk elasticities. AB - The abundance and cytoplasmic organization of keratin filaments enables them to contribute to the maintenance of structural integrity in epithelial tissues. Co polymers of the type II keratin 8 and type I keratin 18 form the major intermediate filament network in simple epithelia. We investigated the mechanical properties of K8-K18 filament suspensions using rheological assays in conjunction with light and electron microscopy. Suspensions of K8-K18 filaments behave like a viscoelastic solid under standard assembly conditions. Bulk elasticity is weakly dependent on deformation frequency but is very sensitive to the concentration (G' approximately C1.5) and size of individual keratin polymers, in agreement with recent models of semiflexible-polymer physics. K8-K18 filaments can self-organize to form a bundled network that exhibits gel-like mechanical properties. In all cases the mechanical properties of the suspensions correlate with the structural features of individual polymers, as seen under light and electron microscopy. Importantly, these bulk viscoelastic properties of K8-K18 filaments are revealed only when interfacial elastic effects are minimized by the application of phospholipids at the air-liquid interface. Suspensions of K5-K14 and vimentin filaments also exhibit interfacial elasticity, which distorts the interpretation of the viscoelastic moduli as determined by standard rheometry. The potential for modulation of mechanical properties through self-organization may be a general property of keratin polymers and contribute to their organization and function in vivo. PMID- 12892726 TI - Concentric intermediate filament lattice links to specialized Z-band junctional complexes in sonic muscle fibers of the type I male midshipman fish. AB - Type I male midshipman fish produce high-frequency hums for prolonged durations using sonic muscle fibers, each of which contains a hollow tube of radially oriented thin and flat myofibrils that display extraordinarily wide ( approximately 1.2 microm) Z bands. We have revealed an elaborate cytoskeletal network of desmin filaments associated with the contractile cylinder that form interconnected concentric ring structures in the core and periphery at the level of the Z bands. Stretch and release of single fibers revealed reversible length changes in the elastic desmin lattice. This lattice is linked to Z bands via novel intracellular desmosome-like junctional complexes that collectively form a ring, termed the "Z corset," around the periphery and within the core of the cylinder. The junctional complex consists of regularly spaced parallel approximately 900-nm-long cytoskeletal rods, or "Z bars," interconnected with slender (3-4 nm) plectin-positive filaments. Z bars are linked to the Z band by plectin filaments and on the opposite side to a dense mesh of desmin filaments. Adjacent Z bands are linked by slender filaments that appear to suspend sarcotubules. We propose that the highly reinforced elastic desmin cytoskeleton and the unique Z band junctions are structural adaptations that enable the muscles' high-frequency and high-endurance activity. PMID- 12892727 TI - Nucleosome gaping supports a functional structure for the 30nm chromatin fiber. AB - The biological functions played by the nucleus of eukaryotic cells and especially those involved in cellular differentiation not only depend on the genomic sequence but also on all the proteins which form the nucleo-protein complex named chromatin. The tridimensional organization of this huge polymer involves many structural levels, the most basic one being the nucleosome. Nucleosomes further organize into the so-called 30nm fiber, which, according to recent works, is likely to be the main functional level of chromatin. We wish here to propose a plausible structure for the 30nm chromatin fiber that could explain its functional role. In our model, silenced chromatin is locked by nucleosome stacking interactions. This is achieved by a conformational transition within the nucleosome core particle (NCP) which allows nucleosomes to stack along two helices without bending the DNA linkers. We used molecular modeling to check that this conformational transition was plausible. Then we proposed to modify the well known two-angle model according to these atomic level results. The emerging picture is an allosteric behavior of the nucleosomes induced by their collective organization within the 30nm chromatin fiber. PMID- 12892728 TI - Lignin distribution in wood cell walls determined by TEM and backscattered SEM techniques. AB - The lignin distribution in cell walls of spruce and beech wood was determined by high-voltage transmission-electron-microscopy (TEM) in sections stained with potassium permanganate as well as by field-emission-scanning-electron-microscopy (FE-SEM) combined with a back-scattered electron detector on mercurized specimens. The latter is a new technique based on the mercurization of lignin and the concomitant visualization of mercury by back-scattered electron microscopy (BSE). Due to this combination it was possible to obtain a visualized overview of the lignin distribution across the different layers of the cell wall. To our knowledge, this combined method was used the first time to analyse the lignin distribution in cell walls. In agreement with previous work the highest lignin levels were found in the compound middle lamella and the cell corners. Back scattered FE-SEM allows the lignin distribution in the pit membrane of bordered pits as well as in the various cell wall layers to be shown. In addition, by using TEM as well as SEM we observed that lignin closely follows the cellulose microfibril orientation in the secondary cell wall. From these observations, we conclude that the polymerisation of monolignols is affected by the arrangement of the polysaccharides which constitute the cell wall. PMID- 12892729 TI - Nucleation and growth of macrofibrils in trichocyte (hard-alpha) keratins. AB - The intermediate filaments (IF) in trichocyte (hard-alpha) keratin form ordered aggregates that are infiltrated by sulfur-rich and tyrosine-rich proteins during fiber development to give a filament-matrix texture, which is stabilized in the later stages by the formation of disulfide linkages. Two polymorphic forms of macrofibril are found in the cortical cells of fine Merino wool. In the first the packing of the IF in the macrofibril is quasi-hexagonal whilst in the second the IF are packed in cylindrical sheets around a central core. In hairs the second type generally predominate. In the present contribution specific models for the mechanisms of nucleation and growth are developed for the two types of macrofibril and their applicability tested by analyzing transmission electron micrographs of wool and hair. Evidence is presented which supports the idea that sheet formation plays an important role in both types of macrofibril assembly and it is suggested that differing intersheet interactions are responsible for the differences between the ortho- and para-types. It is shown that the increase in IF tilt with radius in the ortho-type can be related to the surface lattice of the IF as determined from X-ray diffraction studies. Two possible types of intersheet interaction in the ortho-type are discussed, the first leading to an increase of around 0.5 degrees in IF tilt per layer and the second leading to a much larger tilt of 9.4 degrees per layer. A crude estimate based on the decrease in visibility of the IF with increasing radius in cross-section yielded a value of 0.35 degrees -0.7 degrees. PMID- 12892731 TI - Immunization with streptozotocin-treated NOD mouse islets inhibits the onset of autoimmune diabetes in NOD mice. AB - In this study, we determined whether a single intraperitoneal injection of NOD islets exposed to streptozotocin (STZ; 5 mmol/l) in vitro could prevent onset of diabetes in female NOD mice. Pre-diabetic female NOD mice were injected with saline or islets exposed to either STZ or citrate buffer alone. Single injection of STZ-exposed islets significantly (P<0.03) decreased the incidence of diabetes in pre-diabetic NOD mice compared to control groups. At 40 weeks of age, the onset of diabetes in NOD mice injected with STZ-treated islets was 16% (3/19) compared to 88% (14/16) in mice that received islets exposed to citrate buffer and 84% (26/31) in those mice injected with saline. Histological examination of the pancreases from normoglycemic mice given STZ-treated islets revealed numerous intact islets devoid of mononuclear cell infiltration while pancreases from control groups contained few intact islets infiltrated with mononuclear cells. This study demonstrates that immunization of pre-diabetic female NOD mice with syngeneic islets exposed to STZ prevents insulitis and onset of autoimmune diabetes. Our data suggest that exposure of islets to STZ may possibly induce the release of soluble antigens and/or cause an antigenic change in pancreatic beta cells that subsequently results in immunization of pre-diabetic NOD mice. PMID- 12892730 TI - Induction of lupus autoantibodies by adjuvants. AB - Exposure to the hydrocarbon oil pristane induces lupus specific autoantibodies in non-autoimmune mice. We investigated whether the capacity to induce lupus-like autoimmunity is a unique property of pristane or is shared by other adjuvant oils. Seven groups of 3-month-old female BALB/cJ mice received a single intraperitoneal injection of pristane, squalene (used in the adjuvant MF59), incomplete Freund's adjuvant (IFA), three different medicinal mineral oils, or saline, respectively. Serum autoantibodies and peritoneal cytokine production were measured. In addition to pristane, the mineral oil Bayol F (IFA) and the endogenous hydrocarbon squalene both induced anti-nRNP/Sm and -Su autoantibodies (20% and 25% of mice, respectively). All of these hydrocarbons had prolonged effects on cytokine production by peritoneal APCs. However, high levels of IL-6, IL-12, and TNFalpha production 2-3 months after intraperitoneal injection appeared to be associated with the ability to induce lupus autoantibodies. The ability to induce lupus autoantibodies is shared by several hydrocarbons and is not unique to pristane. It correlates with stimulation of the production of IL-12 and other cytokines, suggesting a relationship with a hydrocarbon's adjuvanticity. The potential to induce autoimmunity may complicate the use of oil adjuvants in human and veterinary vaccines. PMID- 12892732 TI - Idiotype-anti-idiotype circuit in non-autoimmune mice after immunization with the epitope and complementary epitope 289-308aa of La/SSB: implications for the maintenance and perpetuation of the anti-La/SSB response. AB - BACKGROUND: Antibodies to La/SSB are usually found in sera of patients with Sjogren's Syndrome (SS) and Systemic Lupus Erythematosus (SLE). Recent work from our laboratory (Mol Med 2002;8:293-305) revealed that an active idiotypic network involving antibodies to epitopes of La/SSB and their anti-idiotypes exist in human sera. The anti-idiotypic antibodies were detected using complementary peptides to B-cell epitopes of the autoantigen. The principle of the complementary peptides is based on the 'molecular recognition' theory. According to this theory, translation of two complementary RNA strands (coding and non coding strand) into protein, generate a pair of peptides, which bind each other with specificity and high affinity. AIM: To investigate antibody production and T cell responses in non-autoimmune-susceptible animal strains which were immunized with the epitope 289-308aa of La/SSB as well as its complementary epitope. MATERIALS AND METHODS: Balb/c mice were immunized with a peptide corresponding to epitope 289-308aa (pep) or its complementary (cpep) peptide (5 animals/group). The sera were tested for the presence of antibodies to pep and cpep as well as for epitope spreading to recombinant human La/SSB and a major B-cell epitope of La/SSB spanning the region 349-364aa. Another group of animals was sacrificed on day 10 and T-cell responses against pep and cpep were evaluated in cells from lymph nodes and spleen. RESULTS: Immunizations with either pep or cpep led to the appearance of antibodies against the immunogen peptide by day 31 which subsequently was followed by antibody production to its complementary peptide by day 55. In two out of five animals immunized with the epitope 289-308aa, a spreading of the immune response to epitope 349-364aa was observed. In the remaining three animals, negative for antibodies to pep349-364, a specific treatment of sera, using cpep349-364 revealed that anti-idiotypic antibodies masked antibodies to pep349-364. In all immunization experiments high T-cell proliferative responses to both pep and cpep peptides were detected. CONCLUSIONS: Complementary peptides to epitopes of La/SSB can be utilized as probes to study the development of an idiotypic-anti-idiotypic network towards the major autoantigen. The ability of pep and cpep peptides to induce both B-cell and T cell responses may provide useful insights into understanding further the initiation and maintenance of autoimmune response and create new tools for therapeutic intervention. PMID- 12892733 TI - Migration of T cells from nearby inflammatory foci into antibody bound tissue: a relay of T cell and antibody actions in targeting native autoantigen. AB - We have reported that binding of antibody to native autoantigen is prerequisite for T cells to target the native antigen in murine autoimmune ovarian disease model (AOD). As a result, ovarian follicles, with target antigen ZP3 (Zona Pellucida 3), are destroyed. In this study, AOD was induced by co-transfer of ZP3 specific CD4(+)T cells and ZP3 antibody. ZP3 CD4(+)T cells, labeled with CFSE, were found to target macrophages in degenerated follicles to form inflammatory foci, which were composed of mainly endogenous CD4(+)T cells (85%). Only endogenous T cells in the foci, with increased CD69(+)expression, further migrated into antibody bound follicles. No F4/80 or MHC II(+)cells were found to co-migrate with the T cells or in follicles. Co-transfer of ZP3 T cell and antibody also induced (1) a transient PMN influx at early stage and (2) a dramatic increase in IL-1 beta expression coincident with the migration in the ovary. These results suggest that ZP3 antibody binding, only in the presence of ZP3 T cells, may cause an inflammatory change in follicles, which attract endogenous T cells from nearby inflammation. Thus, through a relay between T cell and antibody mediated mechanisms, native autoantigen is targeted and destroyed. This mechanism may explain the requirement of antibody in several T cell mediated autoimmune diseases. PMID- 12892734 TI - Infiltration of the inflamed eye by NKT cells in a rat model of experimental autoimmune uveitis. AB - NK and NKT cells play important roles in both the pathogenesis and regulation of autoimmune disease. In this study, NK and NKT cells in the periphery and in the inflamed eyes of Lewis rats developing uveitis were quantified and evaluated for function. The number and phenotype of NK/NKT cells in the inflamed eyes and spleens were determined. The cytotoxic and suppressive activity of the eye infiltrating NK/NKT cells was also studied. In rats with interphotoreceptor retinoid-binding protein (IRBP)-induced uveitis, the number of NK/NKT cells declined in the periphery, but increased in the eyes. The eye-infiltrating NK/NKT cells inhibited cytokine production by autoreactive T cells in vitro and had a weak cytolytic effect on NK-sensitive YAC-1 cells and NKT-sensitive U937 cells in vitro. Our study shows that the number of NK and NKT cells decreases in the periphery of rats with autoimmune uveitis. It is discussed whether the changing number of NK/NKT cells allows the escape from immune surveillance of uveitogenic (i.e. autoreactive) T cells and the development of intraocular inflammation, and the increased infiltration of the inflamed eye by suppressive NK/NKT cells favors rapid recovery from disease. PMID- 12892735 TI - Linear epitopes of two different autoantigens-La/SSB and myelin basic protein- with a high degree of molecular similarity, cause different humoral immune responses. AB - The region 147-154aa of La/SSB presents 83% sequence similarity with the 139 146aa region of the human myelin basic protein (MBP). The aim of this study was to investigate the prevalence and significance of antibodies against both epitopes in sera from patients with systemic autoimmune diseases, and to compare the humoral responses produced after rabbit immunization. Peptides 147-154aa of La/SSB and 139-146aa of the MBP were attached on tetrameric sequential oligopeptide carriers and used for immunizations of New Zealand White rabbits. Antibodies to immunizing peptides, as well as to the peptides corresponding to other previously defined La/SSB epitopes (289-308aa, 349-364aa), to the intact human MBP (hMBP) and to the recombinant human La/SSB (rechLa) proteins, were identified using specific ELISA assays. Sera from 45 patients with Sjogren's syndrome (pSS), 49 with Systemic Lupus Erythematosus (SLE), and 18 with Rheumatoid Arthritis (RA) were tested against the two peptides and the hMBP. Twenty-two per cent of sera from pSS patients, 27% of SLE patients, and none from RA sera reacted with the La epitope; 27% from pSS sera, 22% of SLE sera, and 17% of RA sera gave a positive reaction against the MBP peptide. Finally, 19% of pSS, 30% of SLE, and 38% of RA sera reacted with the hMBP. Thirty-five days after immunization of rabbits with the La epitope, antibodies were produced against all three La/SSB peptides, the MBP peptide, and the hMBP and rechLa proteins. Rabbits immunized with the MBP peptide produced antibodies against the immunizing peptide and the mimicking peptide of La shortly after immunization, whilst antibodies against the other La epitopes and the two intact proteins were produced later. Inhibition experiments in rabbit sera with high reactivity against hMBP, using the MBP peptide as inhibitor, revealed that 80% of serum reactivity was abolished. In conclusion, a significant proportion of human autoimmune sera reacted with both La and MBP derived peptides, as well as with hMBP. La 147-154aa peptide, when used for animal immunizations, induces a fast epitope spreading involving both La and MBP. In contrast, the mimicking MBP epitope induces a delayed response against the other La epitopes. Thus, despite the fact that these peptides present molecular similarity, they induce different immune responses. PMID- 12892736 TI - Tumor necrosis factor-alpha promoter polymorphisms in Mexican patients with rheumatic heart disease. AB - The major histocompatibility genes (MHC) have been associated with the genetic susceptibility to rheumatic heart disease (RHD). Results have been inconsistent and new genes located on the MHC region such as tumor necrosis factor (TNF-alpha) need to be analyzed. TNF-alpha polymorphisms (positions -238 and -308) were determined in 87 RHD Mexican Mestizo patients and 101 healthy controls. Patients were classified into mitral valve damage (MVD) and multivalvular lesion (MVL) categories. TNF-238 G allele and GG genotype were increased in patients when compared to healthy controls (pC=0.001, OR=14.1 and pC=0.003, OR=14.1, respectively). Also, decreased frequencies of TNF-238 A allele (pC=0.001) and AG genotype (pC=0.003) were found. TNF-308 polymorphism analysis showed increased frequencies of T2 (A) allele (pC<10(-3), OR=10.8) and T1T2 (AG) genotype (pC<10( 3), OR=9.85) and decreased frequencies of T1 (G) allele and T1T1 (GG) genotype (pC<10(-3)). When comparing valvular damage to healthy controls, patients with MVD showed increased frequencies of -238 GG (pC=0.03, OR=ND), -308 T1T2 (AG) (pC<10(-3), OR=14) and -308 T2 (A) (pC<10(-3), OR=11.7). Also, this group showed decreased frequencies of T1 (G) allele and T1T1 (GG) genotype (pC<10(-3)). Patients with MVL presented increased frequency of -308 T2 (A) allele (pC=0.0003, OR=8.65) and decreased frequencies of -308 T1 (G) allele and -308 T1T1 (GG) genotype (pC=0.0003 and pC=0.006, respectively). Distribution of -238 and -308 polymorphisms were similar between MVD and MVL. The data demonstrate that RHD is associated with TNF-alpha polymorphisms in the Mexican population; however, these polymorphisms do not have relation with the valve damage. PMID- 12892737 TI - Cluster analysis of human autoantibody reactivities in health and in type 1 diabetes mellitus: a bio-informatic approach to immune complexity. AB - Informatic methodologies are being applied successfully to analyze the complexity of the genome. But beyond the genome, the immune system reflects the state of the body in health and disease. Traditionally, immunologists have reduced the immune system, where possible, to one-to-one relationships between particular antigens and particular antibodies or T-cell clones. Autoimmune diseases, caused by an immune attack against a body component, are usually investigated by following the response to single self-antigens. In this study, we apply informatics to analyze patterns of autoantibodies rather than single species of autoantibodies. This study was designed not to replace traditional approaches to immune diagnosis, but to test whether meaningful patterns of autoantibodies might exist. Using an unbiased solid-phase ELISA antibody test, we detected serum IgG and IgM antibodies in the sera of 20 healthy persons and 20 persons with type 1 diabetes mellitus binding to an array of 87 different antigens, mostly self-antigens. The healthy subjects manifested autoantibodies to a variety of self-antigens, many known to be associated with autoimmune diseases. We investigated the patterns of these autoantibodies using a coupled two-way clustering algorithm developed for analyzing data from gene arrays. We now report that the reactivity patterns of autoantibodies to particular subsets of self-antigens exhibited non-trivial structure, which significantly discriminated between healthy persons and persons with type 1 diabetes. The results show that despite the wide prevalence of autoantibodies, the patterns of reactivity to defined subsets of self-antigens can provide information about the state of the body. PMID- 12892738 TI - Low frequency of anti-SLA/LP autoantibody in Japanese adult patients with autoimmune liver diseases: analysis with recombinant antigen assay. AB - Anti-soluble liver antigen/liver pancreas (SLA/LP) autoantibody has been proposed to be one of the autoantibodies characterizing autoimmune hepatitis (AIH). Recently, one of the autoantigens to anti-SLA/LP was identified as a UGA suppressor tRNA-associated protein. Although the function of this protein remains unknown, the recombinant protein has been prokaryotically expressed. Using this protein as an antigen, a recombinant immunoassay for anti-SLA/LP autoantibody has been established and the frequency and significance of this autoantibody have been discussed in European countries. So, in the present study, we investigated anti-SLA/LP autoantibodies in Japanese patients with autoimmune liver diseases using the recombinant antigen ELISA and Western blot assay. Seventy-five patients with AIH type 1, 5 with AIH type 2, 46 with primary biliary cirrhosis, 10 with primary sclerosing cholangitis, 47 with chronic hepatitis C, 48 with systemic lupus erythematosus, 3 with cryptogenic hepatitis, and 40 normal controls were the subjects of the present study. Anti-SLA/LP autoantibodies were detected in only 5 of 75 (6.7%) patients with AIH type 1, but in none of the other 159 patients or 40 normal controls. The clinicopathologic features of anti-SLA/LP positive AIH type 1, including carriers of HLA DR locus variations, were not significantly different from anti-SLA/LP-negative patients except for the mortality rate. Anti-SLA/LP autoantibody was detected at a low frequency in Japanese patients with AIH type 1 and did not significantly influence clinical features. However, since it has high disease-specificity to AIH type 1, further analysis of SLA/LP may contribute to help clarify the pathogenesis of AIH type 1. PMID- 12892739 TI - Expression of glucocorticoid induced TNF receptor family related protein (GITR) on peripheral T cells from normal human donors and patients with non-infectious uveitis. AB - The expression of the glucocorticoid induced TNF receptor family related gene (GITR) in subsets of T lymphocytes from human peripheral blood was studied. In normal human peripheral blood mononuclear cells, GITR expression on the resting CD4+ T cells was low but markedly increased after activation. The percentage of GITR+ T cells in the CD4+CD25+ T cell subpopulation (15.1%) was significantly higher than that in the CD4+CD25- T cell subpopulation (5.2%, P<0.01), suggesting a preferential co-expression of GITR with CD25. In a group of patients with non infectious uveitis, a proposed T helper cell mediated autoimmune ocular disease, the GITR expression on the CD4+ T cells in both the active patients (34.5%) and the inactive patients (19.6%) was significantly higher as compared to that in the normal donors (10.7%; P<0.01 vs. active, P<0.05 vs. inactive). This increased GITR expression in T cells was only seen in the CD4 positive T helper cell subpopulation but not in the CD4 negative T cell subpopulation. GITR expression on the CD4+ T cells decreased when the patients became clinically quiescent. Therefore, GITR is an activation marker for the CD4+ T cells and preferentially co-expressed with CD25 on the CD4+ T cells in human peripheral blood. Its expression correlates with the clinical course of non-infectious uveitis. PMID- 12892740 TI - Identification of a scavenger receptor homologue on nonspecific cytotoxic cells and evidence for binding to oligodeoxyguanosine. AB - In mammals scavenger receptors (SR) are expressed by monocytic-macrophage lineage cells and B-cells. Studies of various teleost species have indirectly demonstrated the presence of SR receptors on phagocytic or endothelial cells by showing the uptake of SR ligands (i.e. derivatised (acetylated) lipoproteins) by these cells. In the present study, nonspecific cytotoxic cells (NCC) were examined for membrane expression of an SR-like protein. Approximately 15-25% of purified NCC expressed scavenger receptor class A (SR-A) demonstrated by binding by a monoclonal (2F8) specific for mouse SR-A (types I, II). Flow cytometric analysis determined that SR binding cells had the same size and 'side scatter' characteristics as NCC. Two colour flow analysis of NCC demonstrated that only a subset of NCC expressed the SR-A-like protein and non-NCC were SR-A negative. Membrane expression of SR on NCC was confirmed by fluorescence microscopy. Analysis of the tissue distribution of SR bearing cells demonstrated that in both catfish and tilapia, SR-A was expressed by NCC in the peripheral blood, spleen and anterior kidney. Experiments were also done to determine if the ligands known to bind mammalian SR-A had a similar specificity for the teleost receptor. Cold competition binding experiments determined that anti-SR-A antibody competed with and reduced biotinylated polyguanosine 20-mer binding to NCC by approximately 40%. Two other types of ligands known to bind (mammalian) SR-A (i.e. polyvinyl sulphate and dextran sulphate) likewise decreased anti-SR-A antibody binding to NCC by 40%. These studies for the first time demonstrated that NCC express the teleost orthologue of mammalian SR-A, suggesting that NCC may participate in physiologic regulation of lipid metabolism in addition to functions of innate immunity. PMID- 12892741 TI - A thraustochytrid protist isolated from Mercenaria mercenaria: molecular characterization and host defense responses. AB - A previously undescribed thraustochytrid protist, designated C9G, was isolated from the gills of a clam, Mercenaria mercenaria, collected from the Bay of Fundy, Canada. Sequence data analysis showed C9G to be related to the clam pathogen QPX, quahog parasite unknown; however, it is not enveloped by secreted mucoid material as is the case for QPX. Clam hemocytes recognized and phagocytized C9G in vitro in the absence of plasma recognition factors. Hemocytes were also capable of killing ingested C9G, as shown by the use of a tetrazolium reduction viability assay. The mechanisms underlying intracellular antimicrobial activity are not yet established, but no detectable cytotoxic reactive oxygen species were generated during phagocytosis of C9G. Clam plasma proteins were shown to inhibit C9G growth at concentrations similar to those in unfractionated hemolymph. PMID- 12892742 TI - Bivalve immunity: comparisons between the marine mussel (Mytilus edulis), the edible cockle (Cerastoderma edule) and the razor-shell (Ensis siliqua). AB - Much of the current knowledge concerning bivalve immunology and immunotoxicology has come from studies on the mussel genus, Mytilus, or from the American oyster, Crassostrea virginica. Following a major oil spill, it was observed that the marine mussel, Mytilus edulis, underwent significant immunosuppression but no oil induced mortalities, while in contrast, mass mortalities were noted in the edible cockle, Cerastoderma edule, and the razor-shell, Ensis siliqua. A study comparing immune cells and functions in these three species was initiated (i) to assess whether M. edulis was a representative model species and (ii) to provide baseline data on immunity in two common species, which had previously received little or no attention in this respect. While all three species shared similar cell types, their lectin-binding and enzyme cytochemistry differed considerably. M. edulis had significantly different proportions of haemocytes binding with the lectins concanavalin A, wheatgerm agglutinin and Helix pomatia agglutinin and stained positive for eight enzymes, compared with only five in C. edule and three in E. siliqua. In terms of immune function, M. edulis haemocytes were much more active in phagocytosis and superoxide generation than haemocytes of the other two species. The results show that immune cells and functions differed extensively in these three closely related species, with M. edulis showing a much higher level of immunological vigour that may be linked to its considerable resilience to adverse environmental conditions. This suggests that M. edulis may not be particularly representative of the bivalves in terms of immune reactivity and that a wider range of species should be included in studies of molluscan immunotoxicology. PMID- 12892743 TI - Cloning and characterisation of a putative ST2L homologue from Atlantic salmon (Salmo salar). AB - The ST2L receptor is a member of the interleukin-1 (IL-1) receptor family and has previously been cloned from human, mouse, rat and chicken. This orphan receptor has no known physiological role but has been implicated in T helper cell type 2 effector function. We describe in this report the cloning and characterisation of a cDNA encoding a homologue of ST2L in Atlantic salmon (Salmo salar). The salmon ST2L cDNA is 2364bp in length and has an open reading frame encoding a polypeptide of 582 amino acids. Similar to other members of the IL-1 receptor (IL 1R) family, the predicted protein has a potential signal peptide, extracellular immunoglobulin-like domains, a short transmembrane region and a characteristic cytoplasmic Toll-IL-1R domain. The predicted protein shows 33% identity and 44% similarity to the chicken ST2L homologue. Phylogenetic analyses cluster the putative salmon ST2L with the chicken and the mammalian ST2L homologues, away from the other members of the IL-1R family. Salmon ST2L is constitutively expressed in brain, white and red blood cells, head kidney, liver, gills and muscle, with highest level of expression in spleen. In vivo stimulation of salmon with lipopolysaccaride does not appear to have a significant effect on expression of the ST2L homologue. PMID- 12892744 TI - Measurement of Crassostrea gigas hemocyte oxidative metabolism by flow cytometry and the inhibiting capacity of pathogenic vibrios. AB - A flow cytometric method to measure the production of oxidative metabolism products was adapted for use with Crassostrea gigas hemocytes. The method is based upon the oxidation, by hydrogen peroxide (H2O2), of intracellular 2',7' dichlorofluorescin (DCFH) to green-fluorescent dichlorofluorescein. Activation of the respiratory burst (RB) was tested using phorbol myristate acetate with no success. By contrast, activation by zymosan particles increased oxidation of DCFH in C. gigas hemocytes, mainly granulocytes, and optimization tests showed a good response with 20 zymosan particles per hemocyte. Anti-aggregant solution, used to prevent hemocytes from clumping during bleeding, inhibited the RB activity measured by DCFH oxidation. The flow cytometric method developed during this work was used to evaluate the DCFH oxidation-inhibiting capacity of four strains of vibrio bacteria, known or suspected to be pathogenic for bivalves. PMID- 12892745 TI - Toxicity of nitric oxide and peroxynitrite to Photobacterium damselae subsp. piscicida. AB - Virulent strains of Photobacterium damselae subsp. piscicida (Pdp) were grown in media with or without glucose supplementation (to enhance polysaccharide capsule formation) and the bactericidal action of nitric oxide (NO) and peroxynitrites was evaluated in a cell-free assay. Treatment with the NO-donor S-nitroso-acetyl penicillamine (SNAP) induced a dose-and time-dependent decrease in Pdp survival. This effect was greater for strains grown without glucose supplementation (C forms) than for their counterparts grown with glucose supplementation (C(+) forms). Addition of superoxide anion (O2(-)) generating systems (Xanthine/Xanthine oxidase, glucose/glucose oxidase) to the culture media further enhanced the bactericidal effect of NO. A similar bactericidal effect, with the same pattern of sensitivity, was observed when C+ and C forms of the bacteria were treated with 3-morpholino-sydonimide hydrochloride (SIN-1), a compound which simultaneously generates NO and O2(-). Addition of superoxide dismutase (SOD) or SOD plus catalase (CAT) did not fully reverse the toxic action of SIN-1 and the bactericidal effect was similar for both C and C(+) forms suggesting that while NO alone is sufficient to cause damage in all strains of the pathogen tested, growth in glucose supplemented medium enhanced protection to reactive oxygen intermediates rather than NO. PMID- 12892746 TI - Structural and functional characterisation of the blood cells of the bivalve mollusc, Scrobicularia plana. AB - Light and electron microscopical studies were carried out in order to characterise the blood cells of the bivalve mollusc, Scrobicularia plana. Three types of haemocytes were recognised: eosinophilic granular haemocytes, basophilic granular haemocytes and basophilic agranular haemocytes. The eosinophilic granulocytes were vesicular and contained large granules whereas the basophilic granulocytes were found to contain small granules and glycogen 'lakes'. The basophilic agranular haemocytes were significantly smaller than the granular haemocytes and had a high nucleus to cytoplasm ratio. Functional characterisation of the blood cells identified activity for the lysosomal enzymes: acid phosphatase, beta-glucuronidase, non-specific esterase and arylsulphatase. There was also a weak staining reaction for phenoloxidase and peroxidase activities. Phagocytosis of Gram-positive bacteria was demonstrated by the haemocytes and antibacterial activity was shown by cell-free haemolymph. Assays to determine release of reactive oxygen species from the haemocytes did not detect any reactive oxygen generation. PMID- 12892747 TI - Molecular cloning of G type lysozyme cDNA in common carp (Cyprinus carpio L.). PMID- 12892748 TI - Molecular cloning and sequencing of the silver chimaera (Chimaera phantasma) interleukin-8 cDNA. PMID- 12892749 TI - Metalloproteinase with factor X activating and fibrinogenolytic activities from Vipera berus berus venom. AB - We have previously shown that Vipera berus berus venom contains several factor X activating enzymes. In the present study we have investigated one of them. The enzyme was separated from venom by gel filtration on Sephadex G-100 superfine and chromatography on agarose HPS-7 and phenyl-agarose. The enzyme is a glycosylated metalloproteinase containing hexoses, hexosamines and neuraminic acid. The purified factor X activating enzyme consists of two equal chains (59 kDa). The specificity studies have shown that enzyme is nonspecific factor X activating proteinase hydrolysing also proteins such as azocasein, gelatin and fibrinogen. The enzyme hydrolyses oxidized insulin B-chain at the positions Ala(14)-Leu(15) and Tyr(16)-Leu(17) but it is inactive on fibrin, plasminogen and prothrombin. We used 8-10 amino acid residues containing peptides, which reproduce the sequence around the cleavage sites in factor X, factor IX and fibrinogen, as potential substrates for enzyme. Cleavage products of peptide hydrolysis were determined by MALDI-TOF MS. The peptide Asn-Asn-Leu-Thr-Arg-Ile-Val-Gly-Gly-factor X fragment was cleaved by enzyme at positions Leu(3)-Thr(4) and Arg(5)-Ile(6). The fibrinogen peptide fragment Glu-Tyr-His-Thr-Glu-Lys-Leu-Val-Thr-Ser was hydrolysed at position Lys(6)-Leu(7). PMID- 12892750 TI - Phenoloxidases in ascidian hemocytes: characterization of the pro-phenoloxidase activating system. AB - The phenoloxidase (PO) activity of the hemocytes lysate supernatant from three ascidians species, assayed by means of 3-methyl-2-benzothiazolinone hydrazone hydrochloride, have been compared. PO-containing hemocytes were identified by a cytochemical reaction and the enzymatic activity measured by a spectrophotometric assay of lysate supernatant from hemocyte populations separated on a discontinuous Percoll density gradient. In Styela plicata, the enzyme appeared to be contained in morula cells only. In Ciona intestinalis, PO activity was shown in univacuolar refractile granulocyte and granular hemocyte. In Phallusia mammillata both compartment cell and granular hemocytes were positive. Enzymatic assay following electrophoretic analysis on polyacrylamide gel electrophoresis (PAGE) or SDS-PAGE indicated that hemocyte lysate presented orthodiphenoloxidase (catecholase) activity. The enzymes from the three species differed in molecular size, activating substances and trypsin sensitivity. PMID- 12892751 TI - Molecular cloning and cDNA sequence analysis of carboxypeptidases A1, A2 and B from the Japanese flounder Paralichthys olivaceus. AB - Although pancreatic serine proteases have been cloned in teleosts, no sequence data are currently available on members of the carboxypeptidase (CP) family. Here, we cloned cDNAs coding for two preproCPAs, corresponding to mammalian preproCPA1 and preproCPA2, and one preproCPB from a pancreatic cDNA library of the Japanese flounder, Paralichthys olivaceus. The activation peptides of flounder proCPs completely retained the sequences for inhibition of enzymatic activity of proCPs just like mammalian proCPs. Of 306-309 amino acids in total, 95 amino acids are completely conserved between bovine CPA1 and CPB and flounder CPs. Notably, amino acid residues for Zn(2+) ligands, catalysis and substrate anchoring are completely conserved between flounder and bovine CPs. Three species of flounder preproCPs are all expressed in the pancreas of first feeding larvae. PMID- 12892752 TI - Palmitoylation is required for membrane association of activated but not inactive invertebrate visual Gqalpha. AB - The invertebrate visual G protein, iGqalpha plays a central role in invertebrate phototransduction by relaying signals from rhodopsin to phospholipase C leading to membrane depolarization. Previous studies have shown reversible association of iGqalpha with rhabdomeric membranes regulated by light. To address the mechanism of membrane association we cloned iGqalpha from a Loligo pealei photoreceptor cDNA library and expressed it in HEK293T cells. Mutations were introduced to eliminate putative sites for palmitoylation at cysteines in positions 3 and 4. Membrane and soluble fractions were prepared from cells where iGqalpha was either activated or maintained in the GDP-bound form, followed by identification of iGqalpha through immunoblot analysis. The wild-type iGqalpha was entirely membrane-bound and shown to be post-translationally modified by palmitoylation. The mutant iGqalpha (C3,4A) was not palmitoylated yet it was found to be membrane associated in the inactive state, however, approximately half of the protein became soluble when activated. These results suggest that palmitoylation is not required for membrane association of iGqalpha in the inactive state but is important in maintaining the stable membrane association of activated iGqalpha GTP. The mechanism by which iGqalpha moves away from the membrane into the cytosol in response to prolonged light-stimulation in the native squid eye appears, therefore, to involve both activation and depalmitoylation processes. PMID- 12892753 TI - Replacement of fish oil with sunflower oil in feeds for Atlantic salmon (Salmo salar L.): effect on growth performance, tissue fatty acid composition and disease resistance. AB - Dietary sunflower oil (SFO) was used to gradually replace fish oil (FO) in six diets (which also contained fish meal) for Atlantic salmon parr (initial mass: 21.7 g). The effect on growth performance, tissue fatty acid profiles and disease resistance was monitored after 63 days. At the conclusion of the trial, no significant differences were detected in growth between any of the feeds. Fatty acid composition of whole carcass, dorsal muscle and liver generally reflected that of the diets. Forty percent of the FO could be replaced by SFO before tissue 22:6n-3 was significantly reduced, although other essential and non-essential fatty acids were more susceptible to change. Significant differences were detected in cumulative mortality of Atlantic salmon challenged with Vibrio anguillarum at the trials conclusion, although this was not correlated to the inclusion level of SFO. Despite the changes observed to the tissue fatty acid profile, there was no significant effect on growth suggesting that SFO is a suitable alternative to FO in diets for Atlantic salmon parr when fish meal is also included. PMID- 12892754 TI - Calcium-regulated GTPase activity in the calcium-binding protein calexcitin. AB - Calexcitin (CE) is a calcium-binding protein, closely related to sarcoplasmic calcium-binding proteins, that is involved in invertebrate learning and memory. Early reports indicated that both Hermissenda and squid CE also could bind GTP; however, the biochemical significance of GTP-binding and its relationship to calcium binding have remained unclear. Here, we report that the GTPase activity of CE is strongly regulated by calcium. CE possessed a P-loop-like structure near the C-terminal similar to the phosphate-binding regions in other GTP-binding proteins. Site-directed mutagenesis of this region showed that Gly(182), Phe(186) and Gly(187) are required for maximum affinity, suggesting that the GTP-binding motif is G-N-x-x-[FM]-G. CE cloned from Drosophila CNS possessed a similar C terminal sequence and also bound and hydrolyzed GTP. GTPase activity in Drosophila CE was also strongly regulated by Ca(2+), exhibiting over 23-fold higher activity in the presence of 0.3 microM calcium. Analysis of the conserved protein motifs defines a new family of Ca(2+)-binding proteins representing the first example of proteins endowed with both EF-hand calcium binding domains and a C-terminal, P-loop-like GTP-binding motif. These results establish that, in the absence of calcium, both squid and Drosophila CE bind GTP at near-physiological concentrations and hydrolyze GTP at rates comparable to unactivated ras. Calcium functions to increase GTP-binding and GTPase activity in CE, similar to the effect of GTPase activating proteins in other low-MW GTP-binding proteins. CE may, therefore, act as a molecular interface between Ca(2+) cytosolic oscillations and the G protein-coupled signal transduction. PMID- 12892755 TI - Characterization of the glutamate dehydrogenase gene and its regulation in a euryhaline copepod. AB - Glutamate dehydrogenase (GDH) plays a key role in the metabolism of free amino acids (FAA) in crustaceans and other metazoans. Glutamate synthesized by GDH via reductive amination is the amino group donor for alanine synthesis and the precursor required for proline synthesis. Since both proline and alanine are important intracellular osmolytes in many marine invertebrates, GDH has been widely implicated as playing a central role in response to hyperosmotic stress in these organisms. We have isolated the gene encoding a GDH homolog from the euryhaline copepod Tigriopus californicus and examined the regulation of GDH under salinity stress. The gene encodes a protein of 557 residues with 76% amino acid identity with Drosophila melanogaster GDH. The gene encodes an N-terminal mitochondrial signal sequence peptide. Only a single intron of 71 bp was found in the GDH gene in T. californicus when genomic sequences and cDNA sequences were compared. The levels of GDH mRNA do not increase during hyperosmotic stress in this copepod. The effects of salt and hyperosmotic stress on GDH enzyme activity were also investigated. GDH activities decrease with increasing NaCl concentrations in in vitro enzyme assays, while live animals exposed to hyperosmotic stress showed no change in GDH enzyme activities. Combined, these results indicate that GDH transcription and enzyme activity do not appear to function in the regulation of alanine and proline accumulation during hyperosmotic stress in T. californicus. The manner in which this important physiological process is regulated remains unknown. PMID- 12892756 TI - Molecular cloning of preprocathepsin E cDNA from the stomach of bullfrog Rana catesbeiana. AB - A cDNA library was constructed from a poly(A)(+) RNA fraction of the gastric mucosa of bullfrog Rana catesbeiana. We cloned a cDNA encoding preprocathepsin E (Pre-Pro-CE) from the library. The present study is the first demonstration of the Pre-Pro-CE cDNA of lower vertebrate such as amphibian. Amino acid sequence deduced from the cDNA was compared with partial amino acid sequence determined by Edman degradation, suggesting that the cDNA comprises an open reading frame encoding a signal peptide (16 amino acids), a pro-sequence (33 amino acids) and a mature protein region (348 amino acids). Two consensus tri-peptide sequences (FDT and VDT) as active site and positions of seven cysteine residues were conserved in this amphibian CE. Although the bullfrog CE was deduced to contain one potential N-linked glycosylation site, its position (Asn(139)-Leu(140)-Thr(141)) was different from that of mammalian CEs. Molecular phylogenetic analysis showed that the bullfrog Pro-CE belongs to the typical Pro-CE group among various aspartic proteinases. PMID- 12892757 TI - IMP-GMP 5'-nucleotidase in reptiles: occurrence in a turtle, a tortoise and three species of snakes. AB - IMP-hydrolyzing activity, which is reactive with goose anti-pig lung IMP-GMP 5' nucleotidase (c-N-II: EC.3.1.3.5) serum, was detected in extracts from liver, heart, kidney, spleen, stomach, skeletal muscle and lung from several species of reptiles: The values found in liver (U/mg protein) of one animal were: 4.5 for an ammono-ureotelic turtle (Trionyx sinensis japonicus); 3.7 for an ureo-uricotelic tortoise (Testudo elegans); 13-23 for three species of uricotelic snakes: Elaphe quadrivirgata, Elaphe conspicillata and Elaphe climacophora. These findings suggest that in the liver of snakes, c-N-II may participate in the production of uric acid as an end product of amino acid metabolism. PMID- 12892758 TI - Differential channelling of liver lipids in relation to susceptibility to hepatic steatosis in two species of ducks. AB - In the human, hepatic steatosis can be associated with an imbalance between synthesis, secretion and storage of hepatic lipids, and exhibits a genetic susceptibility. The effect of overfeeding on hepatic lipid channelling was investigated in two genotypes of ducks that differ in their susceptibility to fatty liver, i.e. the common duck, Anas platyrhynchos, and the Muscovy duck, Cairina moschata. Before overfeeeding, the Muscovy duck exhibited a lower subcutaneous adiposity and a higher muscular development, whereas hepatic composition was similar in both genotypes (>5% lipids and triglycerides accounting for 6-10% lipids). In the plasma lipoprotein profile, HDL predominated (5.5-7.8 g/l) over VLDL (0.09-0.25 g/l) and LDL (0.65-1.06 g/l). All lipid and lipoprotein concentrations were lower in the Muscovy duck. In response to overfeeding, the Muscovy duck exhibited a higher degree of hepatic steatosis (62 vs. 50% lipids), and a lower increase in adiposity and in the concentration of plasma triglycerides (6-fold vs. 10-fold) and VLDL (23-fold vs. 34-fold). Thus, certain genotypes may be more responsive to the dietary induction of fatty liver because of a less efficient channelling of hepatic lipids towards secretion into plasma and adipose storage, and the duck may represent a suitable model in which to study the development of hepatic steatosis and its pathogenesis. PMID- 12892759 TI - Myonuclear apoptosis occurs during early posthatch starvation. AB - Apoptosis is a naturally occurring process; it is important for the final shape and size of developing tissues, and it is characterized by some morphological features such as plasma membrane blebbing, nuclear breakdown, chromosomal fragmentation and apoptotic bodies followed by phagocytosis. The objective of the study was to evaluate the occurrence of apoptosis in chickens immediately posthatch under fed and starved conditions. Male broiler chickens were or were not provided feed for the first 3 days posthatch. Chickens were killed immediately after hatch, at 1 day of age, at 2 days of age and at 3 days of age. The Pectoralis thoracicus was removed, fixed, dehydrated, cleared and embedded in paraffin. Muscle sections were labeled using terminal deoxynucleotidyl transferase (TdT)-mediated dUTP Nick-End Labeling (TUNEL) for detection of apoptotic nuclei. Body weights were lower (P<0.05) in the starved compared to the fed group at 2 and 3 days posthatch. Myofiber cross-sectional area was only smaller (P<0.05) in the starved compared to the fed birds at 3 days posthatch. TUNEL-positive nuclei were present at all days for the fed and starved groups. The proportion of TUNEL-positive nuclei was higher (P<0.05) for the starved group at day 2 and day 3 posthatch compared to the fed group at 3 days posthatch. Apoptosis is a mechanism that contributes to the smaller myofiber size observed at 3 days posthatch. PMID- 12892760 TI - Characteristics of NADH-dependent lipid peroxidation in sarcoplasmic reticulum of white shrimp, Litopenaeus vannamei, and freshwater prawn, Macrobrachium rosenbergii. AB - The iron-catalyzed NADH-dependent lipid peroxidation system in sarcoplasmic reticulum (SR) of cultured white shrimp, Litopenaeus vannamei and freshwater prawn, Macrobrachium rosenbergii was characterized. Production of thiobarbituric acid reactive substances was used to measure the activity of lipid peroxidation. In both species, the system preferred NADH to NADPH as the reducing agent. Lipid peroxidation activities of SR from both species increased when reaction temperatures increased from 6 to 26 degrees C. At 66 degrees C, the reaction was no longer NADH-dependent. Acidic pH amplified the lipid peroxidation activity. Sarcoplasmic reticular lipid peroxidation activity in white shrimp was always greater than in freshwater prawn. Fatty acid composition of SR lipids could be a major factor for this outcome. The proportion of n-3 highly unsaturated fatty acids, such as C20:5 and C22:6, in sarcoplasmic reticular lipids of white shrimp was twice of that in freshwater prawn. The results of this study provide important tools required for anti-oxidative nutrient study at sub-cellular level. PMID- 12892761 TI - Lutein-based plumage coloration in songbirds is a consequence of selective pigment incorporation into feathers. AB - Many birds obtain colorful carotenoid pigments from the diet and deposit them into growing tissues to develop extravagant red, orange or yellow sexual ornaments. In these instances, it is often unclear whether all dietary pigments are used as integumentary colorants or whether certain carotenoids are preferentially excluded or incorporated into tissues. We examined the carotenoid profiles of three New World passerines that display yellow plumage coloration-the yellow warbler (Dendroica petechia), common yellowthroat (Geothlypis trichas) and evening grosbeak (Coccothraustes vespertinus). Using high-performance liquid chromatography, we found that all species used only one carotenoid-lutein-to color their plumage yellow. Analyses of blood carotenoids (which document those pigments taken up from the diet) in two of the species, however, revealed the presence of two dietary xanthophylls-lutein and zeaxanthin-that commonly co-occur in plants and animals. These findings demonstrate post-absorptive selectivity of carotenoid deposition in bird feathers. To learn more about the site of pigment discrimination, we also analyzed the carotenoid composition of lipid fractions from the follicles of immature yellow-pigmented feathers in G. trichas and D. petechia and again detected both lutein and zeaxanthin. This suggests that selective lutein incorporation in feathers is under local control at the maturing feather follicle. PMID- 12892762 TI - Pressure effects on the GTPase activity of brain membrane G proteins of deep living marine fishes. AB - In marine fishes, heterotrimeric guanyl nucleotide binding proteins (G proteins), which couple cell surface membrane receptors to their effector elements, are sensitive to hydrostatic pressure. The intrinsic high affinity GTPase activity of the alpha subunits of G proteins in three signaling systems coupled to adenylyl cyclase, the A(1) adenosine receptor, the muscarinic cholinergic receptor and the beta-adrenergic receptor, was tested at pressures up to 340 atm. Brain membrane preparations from four members of the deep-sea teleost fish family Macrouridae were studied. Coryphaenoides armatus, C. filifer, C. rupestris and Macrourus berglax have depth distributions which together span 100-5810 m. Increased pressure inhibited basal GTPase activity only in M. berglax, which of the four species has the shallowest center of abundance. Increased hydrostatic pressure did not alter the response of GTPase activity to the beta-adrenergic receptor agonist isoproterenol. Increased pressure decreased the stimulation of GTPase activity by the A(1) adenosine receptor agonist cyclopentyladenosine (CPA) in C. armatus and M. berglax, and by the muscarinic cholinergic receptor agonist carbamyl choline in C. armatus, C. filifer and M. berglax. Decreased agonist stimulation of the GTPase activity at elevated pressure may result from pressure induced changes in conformational states or inhibition of agonist binding. The binding of the non-hydrolyzable GTP analog guanosine 5'-[gamma-thio]triphosphate (GTP[S]) in response to CPA was determined at 5 degrees C and atmospheric pressure. Six macrourid species and a morid were studied. The halftime (t(1/2)) values for GTP[S] binding, ranging from 20.8 to 40.9 min, are similar to values previously reported for two other cold-adapted fishes. PMID- 12892763 TI - Citrate synthase and pyruvate kinase activities during early life stages of the shrimp Farfantepenaeus paulensis (Crustacea, Decapoda, Penaeidae): effects of development and temperature. AB - Energy metabolism in early life stages of the shrimp Farfantepenaeus paulensis subjected to temperature reduction (26 and 20 degrees C) was determined using the activities of citrate synthase (CS) and pyruvate kinase (PK). At both temperatures, weight-specific activity of CS decreased throughout the ontogenetic development from protozoea II (PZ II) to postlarva XII-XIV (PL XII-XIV). PK activity reached a pronounced peak in PL V-VI, followed by a further decrease in PL XII-XIV. Temperature reduction produced variation in oxygen consumption rates (QO(2)), ammonia-N excretion and in enzyme activities. Ammonia-N excretion was higher at 20 degrees C in mysis III (M III), PL V-VI and PL XII-XIV, resulting in substantially lower O:N ratios in these stages. QO(2) was increased in protozoea II (PZ II) and mysis I (M I) at 26 degrees C, while no difference in QO(2) was detected in the subsequent stages at either temperature. This fact coincided with higher CS and PK activities in M III, PL V-VI and PL XII-XIV at 20 degrees C compared with 26 degrees C. Regressions between individual enzyme activities and dry weight exhibited slope values of 0.85-0.92 for CS and 1.1-1.2 for PK and temperature reduction was reflected by higher slope values at 20 than at 26 degrees C for both enzymes. Weight-specific CS activity was positively correlated with QO(2) at 20 and 26 degrees C, and may thus be used as an indicator of aerobic metabolic rate throughout the early stages of F. paulensis. The variation in enzyme activities is discussed in relation to possible metabolic adaptations during specific ontogenetic events of the F. paulensis life cycle. Here, the catalytic efficiency of energy-metabolism enzymes was reflected in ontogenetic shifts in behaviour such as larval settlement and the adoption of a benthic existence in early postlarvae. In most cases, enhanced enzyme activities appeared to counteract negative effects of reduced temperature. PMID- 12892765 TI - Improved outcome with novel device for low-pressure PTCA in de novo and in-stent lesions. AB - PURPOSE: Complex lesion morphology requiring the use of high pressure to effect lumen expansion and in-stent restenosis (ISR) remain two indications that challenge conventional PTCA balloons. We report on a new PTCA device that is designed to provide precise, low-pressure dilatation of both de novo and in-stent lesions. METHODS: The FX miniRAIL catheter (FX) has an integral wire positioned external to a dilating balloon and a short, 12-mm guidewire lumen distal to the balloon. The balloon inflates against the guidewire and the external wire to prevent slippage and to introduce high focal longitudinal stresses at low inflation pressures. In this initial study, the FX was used in 37 lesions (25 de novo, 12 in-stent; vessel reference diameter=2.73+/-0.49 mm) in 30 patients. A stepwise inflation protocol and QCA were used to determine the balloon pressure at which the stenosis was resolved (stenosis resolution pressure, SRP). RESULTS: All lesions (100%) were easily reached, crossed and dilated without complication. The SRP was 4.5+/-2.9 atm, and no balloon slippage was observed. Residual stenosis after FX was 26.39+/-13.29%. Minor dissections (Types A and B) were observed in eight lesions (21.6%). Target lesion revascularization (TLR) and target vessel revascularization (TVR) at follow-up (8.1+/-1.5 months) were 8.3% and 12.5%, respectively. CONCLUSION: The design of the FX is versatile and appears to provide for a safe, effective and improved low-pressure PTCA technique in de novo and in-stent lesions. PMID- 12892766 TI - Predictors of late cardiac events following treatment with Sr-90 beta-irradiation for instent restenosis. AB - BACKGROUND: Intracoronary radiation therapy (IRT) with Sr-90 using the Novoste Beta-Cath system has been shown to be an effective therapy for instent restenosis (ISR), but the temporal occurrence of cardiac events and the predictors of late complications require further investigation. METHODS: We analyzed the demographics, lesion characteristics and clinical outcomes of 138 consecutive patients with ISR treated with IRT from September 1998 to March 2002. Major adverse cardiac events (MACE) were defined as death, myocardial infarction (MI) or target vessel revascularization (TVR). Characteristics of early (< or =8 months) and late (>8 months) failures were analyzed. RESULTS: Thirty-two (23.1%) of 138 patients had MACE on follow-up; 25% (8/32) of failures occurred late after treatment with IRT. A comparison of the clinical and angiographic profile of early and late failures using univariate analysis indicates no correlations to late failure following IRT. Duration to failure after IRT was 14.25+/-3.69 months in the late group compared to 4.63+/-2.86 months in the early group (P<.001). CONCLUSIONS: Late MACE after IRT with Sr-90 for ISR occur beyond the traditional period for clinical restenosis in 25% of cases and are difficult to predict. Further study is warranted to identify patients at risk for the development of late complications after IRT. PMID- 12892767 TI - Acute procedural complications and in-hospital events after percutaneous coronary interventions: eptifibatide versus abciximab. AB - BACKGROUND: Glycoprotein IIb/IIIa antagonists reduce peri-angioplasty ischemic complications and improve in-hospital outcome in patients undergoing percutaneous coronary interventions (PCI). Prior studies have demonstrated favorable results with both eptifibatide and abciximab. The purpose of this study was to assess whether there are any differences in rates of acute procedural complications and in-hospital events with the use of these two agents. METHODS: A retrospective review of 359 elective PCIs from June 1998 to August 2000 identified 152 PCIs treated with eptifibatide (bolus 180 microg/kg, infusion 2 microg/kg/min for 12 48 h) and 205 PCIs treated with abciximab (bolus 0.25 mg/kg, infusion 10 microg/min for 12 h). All patients received IIb/IIIa antagonists at the initiation of the intervention. RESULTS: The clinical demographics, the angiographic morphology, the indications, and the procedural details were similar in both groups. In the eptifibatide group, the maximum ACT was lower (235+/-45 vs. 253+/-40, P<.0001). The incidence of major procedural and in-hospital events was compared. Eptifibatide and abciximab had similar rates of major complications (death or myocardial infarction) (1.4% vs. 2.9%), repeat PTCA (3.4% vs. 1.9%), and major bleeding (3.3% vs. 4.3%). CONCLUSIONS: Eptifibatide is comparable to abciximab in regards to acute procedural complications and in-hospital events after PCI. PMID- 12892768 TI - Beta-radiation therapy for long lesions in native coronary vessels: a matched comparison between de novo and in-stent restenotic lesions. AB - OBJECTIVE: The purpose of this study was to evaluate effectiveness and to compare clinical outcome of intracoronary beta-radiation to treat long lesions (>20 mm) in patients with de novo stenosis vs. patients with in-stent restenosis (ISR). METHODS: A matched comparison of 44 patients with 63 de novo lesions and 48 patients with 63 ISR lesions (>20 mm) treated with intracoronary beta-radiation was performed. RESULTS: Stents were implanted in 65.1% of de novo and 19% of ISR lesions (P=.001). Radiation doses delivered were 17.2+/-3.0 vs. 20.3+/-3.0 Gy at 2 mm from the source center for de novo and ISR lesions. There was no difference in the incidence of in-hospital events. Clinical follow-up at 16.4+/-6.7 months showed no difference in major adverse cardiac events (MACE) between de novo and ISR patients (27.3% vs. 25%, P=.8). Late total occlusions (LTOs) occurred in eight patients (four in each group) treated with stents at the time of radiation and after discontinuation of ticlopidine. By multivariate analysis, stent implantation was the only predictor of late occlusions (OR 8.25, 95% CI 1.73 38.46, P<.008). Restenosis rates were similar for de novo and ISR lesions (29.3% vs. 23.2%, P=.46), as well as target lesion revascularization (TLR) and target vessel revascularization (TVR) rates (22.7% vs. 22.9% and 29.5% vs. 29.2%, respectively). CONCLUSIONS: Intracoronary beta-radiation gives comparable results when used to treat de novo or ISR lesions provided new stent implantation can be avoided. Long-term combined antiplatelet therapy is mandatory for patients who receive new stents at the time of radiation treatment. PMID- 12892769 TI - Antioxidants attenuate atherosclerotic plaque development in a balloon-denuded and -radiated hypercholesterolemic rabbit. AB - BACKGROUND: Oxidation of lipoproteins is considered to be a key contributor to atherogenesis. Antioxidants are potential antiatherogenic agents because they can inhibit lipoprotein oxidation. Radiation has been shown to increase oxidative stress leading to increased atherogenesis. This study is designed to test the potential of antioxidants to inhibit atherosclerotic plaque progression in balloon-denuded and -radiated rabbits. METHODS AND RESULTS: Two groups of New Zealand white rabbits (n=36) were fed with 1% cholesterol diet (control diet) or with 1% cholesterol diet containing a mixture of various antioxidants for 1 week. Iliac arteries in all the animals were balloon denuded and continued to fed with 0.15% cholesterol diet or 0.15% cholesterol diet containing antioxidants (antioxidant diet). Four weeks after balloon denudation one iliac artery in 12 animals from each group was radiated and all the animals were continued to be fed with the same diet. Four weeks after radiation animals were sacrificed and morphometric analysis of iliac arteries (n=12) in nonradiated and radiated animals were performed. Plaque area (PA) in the rabbits that were fed with cholesterol diet is 0.2+/-0.12 mm2, and it is increased by 2.75-fold (P<.05) in the radiated arteries of animals fed with cholesterol diet. Plaque area in the animals fed with antioxidant diet is 50% less then the one in the animals fed with cholesterol diet. Similarly, plaque area in radiated arteries of the animals fed with antioxidant diet is 50% less then the animals fed with cholesterol diet. CONCLUSION: Antioxidants significantly attenuate atherosclerotic plaque progression in balloon-injured and -radiated hypercholesterolemic rabbits. PMID- 12892770 TI - Embolization of experimental aneurysms using a heparin-loaded stent graft with micropores. AB - PURPOSE: For percutaneous transluminal angioplasty (PTA), a heparin-loaded stent graft, composed of a commercially available metallic stent with a microporous and surface-modified thin film, has been developed. Early controlled endothelialization is promoted by a regular array of micropores produced by an excimer laser ablation technique. Early thrombus is prevented by a drug delivery system established by impregnation of photoreactive gelatin with heparin. Our stent grafts were used for embolization of experimental carotid aneurysms with an autologous external jugular vein patch in dogs. MATERIALS AND METHODS: At 1 month after formation, the aneurysms were occluded with stent grafts. Affected arteries were removed with the aneurysms, immediately (two aneurysms in one dog), 1 week (four aneurysms in two dogs), 1 month (three aneurysms in two dogs) and 3 months (four aneurysms in two dogs) after embolization, and were studied histologically to evaluate patency and endothelialization over the intraluminal surface of the thin film. RESULTS: Treated carotid arteries were all patent with occluded aneurysms completely at any periods. Even at 1 week after embolization, endothelialization was confirmed on the surface of the stent graft on the lumen side. At 1 and 3 months, all treated aneurysms with enough patent parent arteries were filled with organized tissues and completely occluded. CONCLUSION: Our developed stent graft appears to be promising for the treatment of aneurysms, especially with respect to immediate termination of blood inflow and early endothelialization in the neck of the aneurysm. PMID- 12892771 TI - Oral rapamycin inhibits growth of atherosclerotic plaque in apoE knock-out mice. AB - INTRODUCTION: Inflammatory and immunological responses of vascular cells are known to play significant roles in atherosclerotic plaque development. Rapamycin with antiinflammatory, immunosuppressive and antiproliferative properties has been shown to reduce neointima formation when coated on stents. This study is designed to test the potential of oral rapamycin to inhibit atherosclerotic plaque development. METHODS: Eight-week-old apoE knock-out mice were fed with 0.25% cholesterol supplemented diet (control diet), control diet containing 50 microg/kg rapamycin (low-dose rapamycin) or 100 microg/kg rapamycin (high-dose rapamycin) for 4 or 8 weeks. Subsets of mice from each group (n=10) were weighed and euthanized. Whole blood rapamycin levels were determined using HPLC-MS/MS, and histological analyses of atherosclerotic lesions in the aortic root were performed. RESULTS: Mice fed with high-dose rapamycin did not gain weight (18.5+/ 1.5 vs. 20.6+/-0.9 g, P=.01). Blood levels of rapamycin 117+/-7 pg/ml were detected in the blood of mice fed with high-dose rapamycin for 8 weeks. The plaque area in mice fed with high dose oral rapamycin is significantly less as compared to control (0.168+/-0.008 vs. 0.326+/-0.013 mm2, P=.001 at 4 weeks; 0.234+/-0.013 vs. 0.447+/-0.011 mm2, P=.001 at 8 weeks). Lumen area was inversely proportional to the plaque area. CONCLUSIONS: The results indicate that oral rapamycin is effective in attenuating the progression of atherosclerotic plaque in the mice. PMID- 12892772 TI - Cellular cardiomyoplasty: a preliminary clinical report. AB - BACKGROUND: Cellular cardiomyoplasty is the method of transplanting myogenic cells into injured myocardium to restore the lost heart muscle cells and to improve ventricular function. METHOD: Three patients, all with a history of coronary heart disease, underwent coronary artery bypass grafting and implantation of autologous satellite cells. A muscle biopsy of 2-4 g from the right vastus lateralis muscle was obtained for satellite cell (myogenic stem cell from skeletal muscle) isolation and proliferation before implanted into the donor's heart. The cells were suspended in serum-free medium and injected into 30 40 sites at and around the ischemic areas just before reversing the hypothermic cardioplegia to eliminate arrhythmia and to improve retention. After recovery, each patient was maintained at the intensive care unit for 3-4 days with ECG monitoring before transferring to the patient floor. RESULTS: All patients survived the procedure with an uneventful recovery and were discharged from the hospital. At 3-4 months follow-up examination, increased left ventricular ejection fraction of 11% (35-46%), 5.4% (40-45.4%) and 1% (40-41%) and decreased left ventricular diastolic diameter of 4, 2 and 9 mm were observed for the patients, respectively. Arrhythmia was not detected during the follow-up evaluation by ECG. Improved perfusion (99mTC-MIBI) and increased metabolic activity (18F-deoxyglucose) were found at the sites of satellite cell implantation. Significant increase of wall thickness and movement at the areas of cell injection was also observed using 2D-echo. CONCLUSION: Cellular cardiomyoplasty using autologous satellite cells is a safe procedure with encouraging beneficial outcomes in patients. PMID- 12892773 TI - Cellular cardiomyoplasty for a patient with heart failure. AB - BACKGROUND: A 73-year-old man with a history of myocardial infarction and hypertension for 5 years suffered heart failure (NYHA III-IV). METHODS: 2D echo indicated hypokinesia at septal, left ventricular anterior wall and apical regions. Coronary angiograms demonstrated 60% stenosis in distal left main and 99% stenosis in proximal and distal left anterior descending coronary arteries (LAD). Both proximal artery and middle left circumflex coronary artery (LC) had 90% stenosis, and diffuse stenosis of right coronary artery (RC) was found. Myocardial perfusion imaging using 99mTc-MIBI indicated defective perfusion of left ventricular apex, anterior wall and septal region and severe reduced perfusion of posterior inferior wall. Myocardial metabolic activities (18F deoxyglucose) also showed comparable reductions. After exposing the heart, LAD, LC, and RC were all completely occluded and bypass procedure could not be completed. Autologous satellite cells were implanted without any complication and the patient had an uneventful recovery. RESULTS: During the first 2 months, he remained in heart failure, and by the third month, he gradually improved and reached NYHA II. At fifth month after the procedure, significant increased ejection fraction (37.1-48.6%) and wall movement with modest reduction of left ventricular systolic diameter (48-45 mm) were observed. Imaging with 18F deoxyglucose showed dramatic improvement in myocardial metabolic activity with similar improvement in myocardial perfusion (99mTc-MIBI). CONCLUSION: This is the first successful case of cellular cardiomyoplasty without any conjunctional procedure for patient with severe coronary heart disease and heart failure. PMID- 12892774 TI - Focused force angioplasty: theory and application. AB - Focused force angioplasty is a technique in which the forces resulting from inflating an angioplasty balloon in a stenosis are concentrated and focused at one or more locations within the stenosis. While the technique has been shown to be useful in resolving resistant stenoses, its real value may be in minimizing the vascular trauma associated with balloon angioplasty and subsequently improving the outcome. PMID- 12892776 TI - Mechanisms of membrane deformation. AB - Membrane traffic requires the generation of high-curvature lipid-bound transport carriers represented by tubules and vesicles. The mechanisms through which membranes are deformed has gained much recent attention. A major advance has been the demonstration that direct interactions between cytosolic proteins and lipid bilayers are important in the acquisition of membrane curvature. Rather than being driven only by the formation of membrane-associated structural scaffolds, membrane deformation requires physical perturbation of the lipid bilayer. A variety of proteins have been identified that directly bind and deform membranes. An emerging theme in this process is the importance of amphipathic peptides that partially penetrate the lipid bilayer. PMID- 12892777 TI - Lipids in endocytic membrane transport and sorting. AB - Protein complexes associated to specific membrane lipids and protein-lipid domains contribute to regulate protein sorting and membrane dynamics in the endocytic pathway. It is also becoming apparent that different lipid territories are distributed along the pathway, and that some lipids segregate into specialised microdomains. PMID- 12892778 TI - Lipids and the exocytotic machinery of eukaryotic cells. AB - The molecular machines that drive protein transport through the secretory pathway function exert their activities on the surfaces of membrane bilayers. It is now clear that the various lipid components of these bilayers play direct and versatile roles in modulating the activity of proteins that either themselves constitute core components of the membrane trafficking machinery, or represent proteins that regulate such core components. PMID- 12892779 TI - Arf and its many interactors. AB - Arf GTP-binding proteins regulate membrane traffic and actin remodeling. Similar to other GTP-binding proteins, a complex of Arf-GTP with an effector protein mediates Arf function. Arf interacts with at least three qualitatively different types of effectors. First, it interacts with structural proteins, the vesicle coat proteins. The second type of effector is lipid-metabolizing enzymes, and the third comprises those proteins that bind to Arf-GTP but whose biochemical or biological functions are not yet clearly defined. Arf interacts with two other families of proteins, the exchange factors and the GTPase-activating proteins. Recent work examining the functional relationships among the diverse Arf interactors has led to reconsideration of the prevailing paradigms for Arf action. PMID- 12892780 TI - Golgins in the structure and dynamics of the Golgi apparatus. AB - Golgins are a family of coiled-coil proteins associated with the Golgi apparatus necessary for tethering events in membrane fusion and as structural supports for Golgi cisternae. Recent work has shown that golgins such as GM130, golgin-45 and p115 bind to Rab GTPases via their coiled-coil domains, and that GM130, rather than being part of a static structural matrix, is in dynamic exchange between the membrane surface and the cytoplasm. Golgins such as bicaudal-D1 and -D2 bind to Rab6, but, rather than tethering membranes together, link vesicles to the cytoskeleton, thus adding a new function for this class of proteins. Other golgins containing the Golgi targeting GRIP domain, rather than binding Rabs, interact with and are recruited to membranes by another class of GTPase, the Arls. Current evidence therefore suggests that golgins function in a variety of membrane-membrane and membrane-cytoskeleton tethering events at the Golgi apparatus, and that all these are regulated by small GTPases of the Rab and Arl families. PMID- 12892781 TI - Insider information: what viruses tell us about endocytosis. AB - Viruses have long served as tools in molecular and cellular biology to study a variety of complex cellular processes. Currently, there is a revived interest in virus entry into animal cells because it is evident that incoming viruses make use of numerous endocytic pathways that are otherwise difficult to study. Besides the classical clathrin-mediated uptake route, viruses use caveolae-mediated endocytosis, lipid-raft-mediated endocytic pathways, and macropinocytosis. Some of these are subject to regulation, involve novel endocytic organelles, and some of them connect organelles that were previously not known to communicate by membrane traffic. PMID- 12892782 TI - The apical compartment: trafficking pathways, regulators and scaffolding proteins. AB - Defects in the trafficking of apical membrane proteins in polarized epithelial cells are often associated with diseases, including cystic fibrosis, Liddle's syndrome, nephrogenic diabetes insipidus and Dubin-Johnson syndrome. In recent years, we have learned much about the specialized apical trafficking pathways in polarized cells. Many laboratories have identified signals that direct proteins within these pathways and have defined protein interactions that mediate specific steps in the sorting and stabilization of these proteins. In addition, many cytosolic proteins, including lipid kinases, GTPases, ATPases and scaffolding/adaptor proteins that lack enzymatic activity, regulate the trafficking of proteins through these pathways. Recent advances in the field include the role of small GTPases, unconventional myosins and lipid kinases in apical endocytosis and transcytosis, and the identification of PDZ proteins that regulate apical membrane trafficking of receptors, transporters and ion channels. PMID- 12892783 TI - Spatial control of exocytosis. AB - During many key biological processes, exocytosis is confined to distinct regions of the plasma membrane. Spatial control of exocytosis correlates with altered membrane skeleton dynamics and assembly of local membrane microdomains. These domains act as local stages for the assembly and the regulation of molecular complexes (targeting patches) that mediate vesicle-membrane fusion. Furthermore, local activation of signaling pathways reinforces formation of these patches and might effect global repositioning of the secretory pathway toward sites of localized exocytosis. PMID- 12892784 TI - Retrograde traffic in the biosynthetic-secretory route: pathways and machinery. AB - In the secretory pathway, the forward (anterograde) membrane flow is compensated by retrograde transport of proteins and lipids. Membrane recycling is required for the maintenance of organelle homeostasis and the re-use of components of the transport machineries for the generation of new transport intermediates. However, the molecular mechanisms and other cellular functions of retrograde traffic are still poorly understood. In recent years, a multitude of protein factors that function in the secretory pathway have been discovered, most of them originally suggested to play a role in forward trafficking. However, in many cases subsequent studies have revealed that these proteins participate (also) in retrograde traffic. It is likely that this shift will continue, reflecting the fact that the two pathways are intimately connected. PMID- 12892785 TI - Protein sorting into multivesicular endosomes. AB - Multivesicular endosomes are important as compartments for receptor downregulation and as intermediates in the formation of secretory lysosomes. Work during the past year has shed light on the molecular mechanisms of protein sorting into multivesicular endosomes and yielded information about the machinery involved in multivesicular endosome formation. Monoubiquitination functions as a signal for sorting transmembrane proteins into intraluminal vesicles of multivesicular endosomes and subsequent delivery to lysosomes. A molecular machinery that contains the ubiquitin-binding protein Hrs/Vps27 appears to be central in this sorting process. Three conserved multisubunit complexes, ESCRT-I, -II and -III, are essential for both sorting and multivesicular endosomes formation. Enveloped RNA viruses such as HIV can redirect these complexes from multivesicular endosomes to the plasma membrane to facilitate viral budding. PMID- 12892786 TI - Death from within: apoptosis and the secretory pathway. AB - Recent studies have highlighted the importance of the secretory pathway in stress induced apoptotic signaling. Sensing stress at the endoplasmic reticulum and Golgi might first trigger recovery mechanisms, followed by apoptosis if repair is unsuccessful. Cleavage of endoplasmic-reticulum- or Golgi-resident proteins can signal repair or apoptosis and promote organelle disassembly during apoptosis. Initiation of apoptosis from the secretory pathway requires components of the death machinery localized to these membranes. Extensive trafficking between compartments of the secretory pathway might allow the cell to integrate signals and to determine the proper response to a particular stress. PMID- 12892787 TI - Cell-cycle-specific Golgi fragmentation: how and why? AB - The Golgi membranes, in the form of stacks of cisternae, are contained in the pericentriolar region of mammalian cells. During mitosis, these membranes are fragmented and dispersed throughout the cell. Recent studies are revealing the significance of pericentriolar position of the Golgi apparatus and mechanism by which these membranes are fragmented during mitosis. PMID- 12892788 TI - Understanding the cell biology of antigen presentation: the dendritic cell contribution. AB - The study of the cell biology of antigen processing and presentation has greatly contributed to our understanding of the immune response. The work of many immunologically inclined cell biologists has also permitted us to gain new insights on cellular mechanisms shared by many cell types. Dendritic cells are master regulators of the immune system and consequently have received a lot of attention in recent years. With the aim of controlling antigen processing and presentation, the solutions used by dendritic cells to respond to environmental changes are numerous and surprising. In the presence of pathogens, dendritic cells regulate strongly their endocytic pathway by interfering with uptake, proteolysis, membrane dynamics and transport in and out of the lysosome to become the most potent antigen-presenting cells known. PMID- 12892789 TI - Modulation of developmental signals by endocytosis: different means and many ends. AB - Recent advances have highlighted the importance of endocytic processes in regulating the activity and distribution of developmental signals. Classically, signalling is downregulated by endocytosis and subsequent trafficking to lysosomes (e.g. Notch, Hedgehog, Roundabout). However, endocytosis can also have a positive role in signalling. For example, endocytosis of Delta, the ligand of Notch, is needed for activation of the signal. In the case of signalling by Hedgehog, endocytic trafficking segregates an inhibitory receptor (Patched) from the positive effector (Smoothened). Endosomes could also be the site where signalling is activated (e.g. transforming growth factor beta). Finally, endocytosis could power the transport of morphogens along epithelia. PMID- 12892790 TI - Staying in aerobic shape: how the structural integrity of mitochondria and mitochondrial DNA is maintained. AB - The structure and integrity of the mitochondrial compartment are features essential for it to function efficiently. The maintenance of mitochondrial structure in cells ranging from yeast to humans has been shown to require both ongoing fission and fusion. Recent characterization of many of the molecular components that direct mitochondrial fission and fusion events have led to a more complete understanding of how these processes take place. Further, mitochondrial fragmentation observed when cells undergo apoptosis requires mitochondrial fission, underlying the importance of mitochondrial dynamics in cellular homeostasis. Mitochondrial structure also impacts mitochondrial DNA inheritance. Recent studies suggest that faithful transmission of mitochondrial DNA to daughter cells might require a mitochondrial membrane tethering apparatus. PMID- 12892791 TI - Peroxisome biogenesis: advances and conundrums. AB - Investigations of peroxisome biogenesis in diverse organisms reveal new details of this unique process and its evolutionary conservation. Interactions among soluble receptors and the membrane peroxins that catalyze protein translocation are being mapped. Ubiquitination is observed. A receptor enters the organelle carrying folded cargo and recycles back to the cytosol. Tiny peroxisome remnants vesicles and tubules - are discovered in pex3 mutants that lack the organelle. When the mutant is transfected with a good PEX3 gene, these protoperoxisomes acquire additional membrane peroxins and then import the matrix enzymes to reform peroxisomes. Thus, de novo formation need not be postulated. Dynamic imaging of yeast reveals dynamin-dependent peroxisome division and regulated actin-dependent segregation of the organelle before cell division. These results are consistent with biogenesis by growth and division of pre-existing peroxisomes. PMID- 12892792 TI - Phagocytosis: latex leads the way. AB - Phagocytosis is the process that cells have evolved to internalise large particles such as mineral debris, which they store, or apoptotic cells and pathogens, which they have the capacity to kill and degrade. However, several important pathogens can suppress these killing functions and survive and multiply within phagosomes, causing disease. Recent advances in phagosome biology have been made possible largely by a model system that uses inert latex beads. The ability to purify latex bead-containing phagosomes has opened the door to allow comprehensive biochemical analyses and functional assays to study the molecular mechanisms governing phagosome function. These approaches have led to unique insights directly relevant for the understanding of the biology of intracellular pathogens and the ways by which they subvert their hosts. PMID- 12892794 TI - Does means of access affect the incidence of small bowel obstruction and ventral hernia after bowel resection? Laparoscopy versus laparotomy. AB - BACKGROUND: Laparotomy for bowel resection is causally related to the development of small bowel obstruction (SBO) and ventral hernia, with incidences approaching 12% to 15% each. This report attempts to define the incidence of these access related complications in a large group of patients undergoing laparoscopic assisted bowel resection (LABR) and open bowel operation (OPEN). STUDY DESIGN: A retrospective cohort of 716 consecutive patients undergoing either LABR (n = 211) or OPEN (n = 505) procedures between January 1995 and July 2000 was identified and selected from a prospective registry. RESULTS: Index LABR (n = 211) and OPEN (n = 505) cases included segmental colectomy in 146 LABR and 408 OPEN patients; subtotal colectomy with or without stoma in 18 LABR and 6 OPEN patients; ileocolectomy in 37 LABR and 85 OPEN patients; and small bowel resection in 10 LABR and 6 OPEN patients. The mean followup periods in the LABR and OPEN groups were 2.71 years and 2.42 years, respectively. The incidence of wound hernia was significantly higher in OPEN cases (n = 65) compared with LABR (n = 5) (p < 0.05). The incidence of surgical repair of ventral hernia was also significantly higher in the OPEN group (28) compared with LABR (4) (p < 0.05). Postoperative SBO requiring hospitalization with conservative management occurred significantly less frequently in LABR patients (n = 4) compared with OPEN patients (n = 31) (p = 0.016). The need for surgical release of SBO was similar between the OPEN and LABR groups (n = 4 versus n = 11). The overall reoperation rate for these two complications was two times higher in the OPEN group than in the LABR group (7.7% versus 3.8%). CONCLUSIONS: The data demonstrate that laparoscopic access for bowel operation significantly reduces the incidence of ventral hernia and SBO rates compared with laparotomy. This reduces the need for readmission to the hospital and additional surgical procedures, providing a potential source of decreased morbidity. It should be considered as a means of cost savings associated with laparoscopic bowel operations. PMID- 12892795 TI - Lymph node surgery in papillary thyroid carcinoma. AB - BACKGROUND: The impact of nodal disease remains controversial in papillary thyroid carcinoma (PTC). STUDY DESIGN: One surgeon treated 159 unselected patients, who were followed up for 1 to 27 years. We present a retrospective analysis with respect to nodal disease. Occult nodal disease was investigated, including metachronous nodal disease (mpN(1)) in primarily node negative patients (pN(0), clinical [c]N(0)). RESULTS: Therapeutic lymphadenectomies, prophylactic lymphadenectomies, or no lymphadenectomy were carried out in 42 (cN(1)), 29 (cN(0)), and 88 (cN(0)) patients, respectively, with stage pN(1) in 41 (98%), in 5 (17%), and in 2 (2.3%) patients, respectively (17% versus 2.3% p < 0.005). Sensitivity and specificity of clinical staging were 85% and 99%, respectively. More frequent prophylactic lymphadenectomy during the study period (p = 0.002) led to a nonsignificant increase in stage pN(1) (26% versus 30%). Immunohistochemistry led to upstaging of only 3% of histologically negative nodes and one (4%) pN(0) patient. Nodal recurrence occurred in 8 of 156 patients (5%) treated for cure, in 12% of pN(1) versus 3% of pN(0) cN(0) tumors (p = 0.009), in 15% of TNM high-versus 3% of low-risk patients (p = 0.006), and in 5% each of patients, younger than 45 and 45 years or more. In TNM high-risk patients, tumor related survival was 50% for stage pN(1) versus 86% for stage pN(0), cN(0) (p = 0.03) (100% and 100% in low-risk patients). CONCLUSIONS: The rate of occult nodal disease might be relatively low, and it does not frequently progress to clinical recurrent disease. Clinical nodal status might be valid for deciding the extent and radicality of node dissection. Prophylactic (central) lymphadenectomy should be carried out without radicality-associated morbidity. Macroscopic nodal disease warrants more rigorous, compartment-oriented lymphadenectomy. There is no rationale for detection of occult disease and micrometastasis by frozen section or immunohistochemistry. PMID- 12892796 TI - Prognostic indicators of outcomes in patients with distant metastases from differentiated thyroid carcinoma. AB - BACKGROUND: Distant metastasis is uncommon in differentiated thyroid cancer and the prognosis is unclear. This study aims to evaluate outcomes and to define independent variables that are associated with tumor-related mortality in patients with distant metastasis from thyroid carcinoma. STUDY DESIGN: A retrospective review of the thyroid cancer research database identified 336 patients with distant metastasis from differentiated thyroid carcinoma treated at a single institution between 1941 and 2000. After excluding patients with local or regional recurrence, distant disease was either the first site of recurrence or was detected at the time of diagnosis of the primary tumor in 242 patients (72%). Patient, tumor, and treatment-related factors were analyzed for their relation to disease-specific survival (DSS) using multivariate Cox regression and the log-rank test. RESULTS: Median survival was 4.1 years and 10-year DSS was 26%. Distant disease was synchronous with the primary diagnosis in 97 of 242 (40%) patients. The site of metastasis was lung only in 103 (43%) patients, bone only in 80 (33%), other sites in 14 (6%), and more than one organ system in 45 (19%). Multivariate analysis identified age 45 years or more, symptoms, site other than lung only or bone only, and no radioactive iodine treatment for the metastasis as predictors of poor outcome with 13%, 11%, 16%, and 12% 10-year DSS, respectively. This compares with age less than 45 years, asymptomatic presentation, metastasis only in the lung or bone, and radioactive iodine treatment with 10-year DSS rates of 58%, 45%, 32%, and 33%, respectively (all p < 0.0001). Radioactive iodine treatment was more often given in patients who were less than 45 years of age, asymptomatic, and with metastasis only in the lung or bone only (p = 0.03, 0.11, 0.01). CONCLUSIONS: Longterm survival is possible in patients with distant metastasis from differentiated thyroid cancer. This retrospective study found that age of 45 years or more, site other than lung only or bone only, and symptoms at the time of diagnosis are associated with poorer outcomes. PMID- 12892797 TI - Lymph node involvement in epithelial ovarian cancer: analysis of 276 pelvic and paraaortic lymphadenectomies and surgical implications. AB - BACKGROUND: The purpose was to determine the factors influencing nodal involvement and topography of pelvic and paraaortic node involvement in ovarian carcinoma. STUDY DESIGN: Between 1985 and 2001, 276 women with epithelial ovarian carcinoma underwent systematic bilateral pelvic and paraaortic lymphadenectomy. RESULTS: The overall frequency of lymph node involvement was 44% (122 of 276). The frequency of pelvic and paraaortic metastases were 30% (82 of 276) and 40% (122 of 276), respectively. The frequency of lymph node metastases according to the stage of the disease (stages I, II, and III + IV) were: 20% (17 of 85), 40% (6 of 15), and 55% (99 of 176), respectively. In patients with stage IA, IB, and IC disease, the rates of nodal involvement were 13% (8 of 60), 33% (4 of 12), and 38% (5 of 13), respectively. None of 15 patients with stage IA grade 1 disease had nodal involvement. None of the 20 patients with mucinous tumors confined to the ovary(ies) (stage I disease) had nodal involvement. When paraaortic nodes were involved, the left paraaortic chain above the level of the inferior mesenteric artery was the most frequently involved site (70 patients, 63%). One of nine patients (11%) with a macroscopic stage I unilateral tumor and paraaortic involvement had contralateral metastases. CONCLUSIONS: Lymphadenectomy should be performed even in patients with stage IA disease. This procedure could be omitted in patients with mucinous apparent stage I disease and stage I grade 1 tumor. Lymphadenectomy should involve the whole pelvic and paraaortic chain up to the level of the left renal vein. A bilateral dissection should be performed even in cases of patients with a unilateral tumor. PMID- 12892798 TI - Percutaneous cholecystostomy in patients with acute cholecystitis: experience of 45 patients at a US referral center. AB - BACKGROUND: Standard treatment for acute cholecystitis is cholecystectomy, but some patients are at high risk for immediate surgery. Percutaneous cholecystostomy might be the procedure of choice in this group. We reviewed the experience of percutaneous cholecystostomy in a large tertiary center population. STUDY DESIGN: We performed a retrospective analysis of patients who underwent percutaneous cholecystostomy, and recorded indications for cholecystostomy, duration of tube placement, clinical outcome, death within 30 days of procedure, complications, bacteriology of aspirated bile, gallbladder contents, and performance of interval cholecystectomy. RESULTS: Forty-five patients (mean age 63 years) had cholecystostomy tubes placed from July 1999 to March 2002. All had confirmed or presumed acute cholecystitis. Mean duration of tube insertion was 54.3 days. Thirty-six patients improved clinically within 5 days. Nine patients died within 30 days; only one death was directly related to gallbladder sepsis. Nine patients subsequently had laparoscopic cholecystectomy, eight had open cholecystectomy, and two had cholecystoenterostomy. Cholecystectomy was planned in another five patients. Cholecystostomy tubes leaked in two patients, blocked in four, and dislodged in one. One patient developed a hemoperitoneum. Bile aspirated at cholecystostomy was culture positive in 12 patients, negative in 16, and not sent or recorded in 17. Twenty-two patients had gallstones, 10 had sludge, 9 had both, and 4 had neither. CONCLUSIONS: In experienced hands, percutaneous cholecystostomy is easy to perform, with low complication and high success rates. It is the procedure of choice in patients with acute cholecystitis unfit for emergency surgery. Patients often improve clinically, so that cholecystectomy can be done electively. PMID- 12892799 TI - Magnitude of serosal changes predicts peritoneal recurrence of gastric cancer. AB - BACKGROUND: Peritoneal dissemination is the most frequent mode of recurrence in patients with gastric cancer. We tried to identify factors that predict peritoneal recurrence with high sensitivity. STUDY DESIGN: Clinical and pathologic data from 587 consecutive patients with gastric cancer were reviewed retrospectively. The stepwise Cox proportional hazards regression model was used to assess the prognostic significance of the magnitude of serosal changes. Multiple stepwise logistic regression analysis was used to determine factors associated with peritoneal recurrence in 375 patients who underwent curative resection. RESULTS: The 5-year survival rate of patients with S2 disease (greatest dimension of macroscopic serosal changes >/= 2.5 cm) was 18%, which was worse than S0 (no serosal changes) and S1 disease (macroscopic serosal changes < 2.5 cm)(p < 0.001). Patients with S0 tumors who underwent curative resection had the best 5-year survival rate. Multivariate analyses indicated that the magnitude of serosal changes was an independent prognostic factor for survival both overall and after curative resection. Logistic regression analysis showed that peritoneal recurrence was more than four times as likely with S2 than with S0 or S1 tumors. The sensitivity for predicting peritoneal recurrence was 79%; the sensitivity of cytologic examination was 38%. CONCLUSIONS: Magnitude of serosal changes is easy to measure intraoperatively and predicts peritoneal recurrence of gastric cancer with greater sensitivity than conventional peritoneal lavage cytology. PMID- 12892800 TI - Zero mortality after 152 consecutive pancreaticoduodenectomies with pancreaticogastrostomy. AB - BACKGROUND: Since 1968, there have been three published reports in the United States literature of 41, 118, and 145 consecutive patients undergoing pancreaticoduodenectomy without mortality. In all of these series, the pancreatic remnant was anastomosed to the jejunum. STUDY DESIGN: This study was designed to review 152 consecutive patients who underwent pancreaticoduodenectomy in whom the pancreatic remnant was anastomosed to the stomach (pancreaticogastrostomy). RESULTS: A total of 152 patients underwent pancreaticoduodenectomy with pancreaticogastrostomy between July 1992 and May 2002. There were 85 men and 67 women with a mean age of 65.7 years (range 31 to 90 years). Of the patients, 87 were less than 69 years of age and 65 were more than 69 years. A total of 114 patients had a malignant neoplasm and the remaining 38 had either cystic neoplasms or benign disease. When the two groups were compared, the patients who were more than 69 years of age had a significantly high incidence of hypertension, previous cancer, atrial fibrillation, and coronary artery disease. In addition, patients more than 69 years of age had a significantly high incidence of jaundice and placement of preoperative stents. Patients more than 69 years of age had significantly less operative time but there was no between-group difference in estimated blood loss, transfusion, number of units transfused, and postoperative length of stay. There was no postoperative mortality [corrected] in this series. Pancreatic leak and fistulae were the most common complications, followed by intraabdominal abscess, wound infection, and delayed gastric emptying. CONCLUSIONS: In this study, 152 consecutive patients underwent pancreaticoduodenectomy with pancreaticogastrostomy without postoperative mortality. Morbidity was mostly because of pancreatic leaks and fistulae, which were successfully treated nonoperatively. With proper selection, careful preoperative preparation, and proper intraoperative conduct of operation, the Whipple procedure can be performed without postoperative mortality. PMID- 12892803 TI - Simultaneous liver and colorectal resections are safe for synchronous colorectal liver metastasis. AB - BACKGROUND: The optimal surgical strategy for the treatment of synchronous resectable colorectal liver metastasis has not been defined. The aims of this study were to review our experience with synchronous colorectal metastasis and to define the safety of simultaneous versus staged resection of the colon and liver. STUDY DESIGN: From September 1984 through November 2001, 240 patients were treated surgically for primary adenocarcinoma of the large bowel and synchronous hepatic metastasis. Clinicopathologic, operative, and perioperative data were reviewed to evaluate selection criteria, operative methods, and perioperative outcomes. RESULTS: One hundred thirty-four patients underwent simultaneous resection of a colorectal primary and hepatic metastasis in a single operation (Group I), and 106 patients underwent staged operations (Group II). Simultaneous resections tend to be performed for right colon primaries (p < 0.001), smaller (p < 0.01) and fewer (p < 0.001) liver metastases, and less extensive liver resection (p < 0.001). Complications were less common in the simultaneous resection group, with 65 patients (49%) sustaining 142 complications, compared with 71 patients (67%) sustaining 197 complications for both hospitalizations in the staged resection group (p < 0.003). Patients having simultaneous resection required fewer days in the hospital (median 10 days versus 18 days, p = 0.001). Perioperative mortality was similar (simultaneous, n = 3; staged, n = 3). CONCLUSIONS: Simultaneous colon and liver resection is safe and efficient in the treatment of patients with colorectal cancer and synchronous liver metastasis. By avoiding a second laparotomy, the overall complication rate is reduced, with no change in operative mortality. Given its reduced morbidity, shorter treatment time, and similar cancer outcomes, simultaneous resection should be considered a safe option in patients with resectable synchronous colorectal metastasis. PMID- 12892806 TI - Prospective randomized study of a protein-based tissue adhesive used as a hemostatic and structural adjunct in cardiac and vascular anastomotic repair procedures. AB - BACKGROUND: The purpose of this study was to determine whether adjunctive use of the bovine serum albumin and glutaraldehyde tissue adhesive BioGlue (BioGlue Surgical Adhesive; CryoLife, Inc) could reduce the rate of anastomotic bleeding in patients undergoing cardiac and vascular repair procedures when compared with a standard repair control. This was a prospective multicenter, randomized, controlled clinical trial conducted in accordance with the IRB at each participating institution. STUDY DESIGN: A total of 151 patients consented to participation and were randomly assigned to standard repair plus BioGlue (n = 76) or standard repair alone (n = 75). These two groups were statistically homogeneous for age, gender, race, procedure, and number of anastomoses. Patients underwent cardiac procedures (n = 49), aortic procedures (n = 105), or peripheral vascular procedures (n = 48). RESULTS: Anastomotic bleeding was significantly reduced in the BioGlue group (18.8% of anastomoses) compared with the control group (42.9% of anastomoses, p < 0.001). Pledget use was reduced in the BioGlue group (26.2%) compared with the control group (35.9%, p = 0.047). Days in the ICU and total days in the hospital were slightly higher in the control group. Adverse event profiles were equivalent between the two groups except for occurrence of neurological defects, which were threefold less in the BioGlue group (p = 0.009). CONCLUSIONS: This study demonstrates that using BioGlue as an adjunct to standard repair methods is safe and significantly reduces the occurrence of intraoperative anastomotic site bleeding in cardiac and vascular repair patients. Using BioGlue along suture lines reinforces anastomoses, thus minimizing pledget use. PMID- 12892809 TI - Natural killer cell activity and distant metastasis in rectal cancers treated surgically with and without neoadjuvant chemoradiotherapy. AB - BACKGROUND: We investigated whether impaired preoperative natural killer (NK) cell activity correlates with asynchronous distant metastasis after curative surgery for rectal cancers. In addition, we examined if preoperative chemoradiotherapy for rectal cancers impairs NK cell activity and contributes to the induction of distant metastasis. STUDY DESIGN: Preoperative NK cell activity was examined in 174 rectal cancer patients. All patients were enrolled in this study and followed until asynchronous distant metastasis occurred. RESULTS: The mean NK activity in patients with stage IV disease (n = 20) was significantly lower than seen in other stages. There were no differences among stage I to stage III patients. In stage III patients, the cumulative distant metastasis-free rate after curative surgery was significantly lower in cases with NK activity of 25% or less than those with more than 25%. Preoperative chemoradiotherapy for stage I to III patients significantly impaired NK cell activity (n = 39), and the metastasis-positive ratio significantly increased among patients with stage II or stage III diseases (n = 30). Multivariate analysis indicated that dichotomized NK cell activity was a significant risk factor that is associated with distant metastasis as well as nodal involvement. CONCLUSIONS: In primary rectal cancers, NK cell activity is not necessarily impaired in accordance with the disease progression. It is considered an important background factor for developing asynchronous distant metastases in stage III rectal cancers. Neoadjuvant chemoradiotherapy impaired NK cell activity in selected patients, suggesting the necessity of concurrent immunotherapy for better outcomes. PMID- 12892810 TI - The nitric oxide donor molsidomine improves survival and reduces hepatocyte apoptosis in cholestasis and endotoxemia. AB - BACKGROUND: Cholestasis and endotoxemia have been demonstrated to cause hepatocyte apoptosis through caspase-mediated pathways. In vitro nitric oxide (NO) donors reduce hepatocyte apoptosis and caspase activation in several models. The nitric oxide donor molsidomine improves survival in an in vivo model of endotoxemia. We tested the effect of molsidomine on survival and hepatocyte apoptosis in a model of obstructive jaundice and endotoxemia. STUDY DESIGN: Sprague-Dawley rats underwent common bile duct ligation on day 1. On day 3, animals were given either 100 mg/kg of molsidomine or an equivalent volume of saline, and 30 minutes later they were given endotoxin 3 mg/kg or 10 mg/kg intravenously. Animals were sacrificed 4 or 16 hours after endotoxin injection. Serum samples were analyzed for alanine aminotransferase and frozen liver samples were analyzed for caspase 3 activity. Paraffin-embedded liver sections were assayed for apoptosis using the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling assay. Survival was measured in a separate experiment in which animals underwent the same protocol, but were given three different doses of endotoxin and were observed for 72 hours before sacrifice. RESULTS: At endotoxin 3 mg/kg, the 72-hour survival in saline-treated animals was 92%, which decreased to 45% at 10 mg/kg and to 29% at 15 mg/kg. All of the molsidomine-treated animals survived all endotoxin doses. Alanine aminotransferase was reduced in molsidomine-treated animals compared with those treated with saline. Apoptosis was attenuated in molsidomine-treated animals. Caspase 3 activity was decreased in molsidomine-treated animals compared with those given saline. CONCLUSIONS: Molsidomine attenuates caspase activation and hepatocyte apoptosis and improves survival after cholestatic endotoxic injury. PMID- 12892811 TI - Poly(ADP-ribose) polymerase inhibition improves postischemic myocardial function after cardioplegia-cardiopulmonary bypass. AB - BACKGROUND: Poly(ADP-ribose) polymerase activation has been shown to contribute to the pathogenesis of myocardial ischemia-reperfusion injury. We hypothesized that a novel poly(ADP-ribose) polymerase inhibitor, INO-1001, provides myocardial protection and improves cardiac function after regional ischemia and cardioplegia cardiopulmonary bypass (CPB). STUDY DESIGN: Pigs were subjected to 30 minutes of regional ischemia by distal left anterior descending coronary artery ligation followed by CPB (60 minutes) with hyperkalemic cardioplegia (45 minutes). The myocardium then was reperfused post-CPB for 90 minutes. After 15 minutes of ischemia, the treatment group (n = 6) received an INO-1001 bolus (1mg/kg) before a continuous infusion (1mg/kg/hour). Control pigs (n = 6) received vehicle solution. Left ventricular pressure was monitored, from which the maximum, positive first derivative of left ventricular pressure over time (+dP/dt) was calculated. Regional myocardial function in the ischemic area was determined by sonomicrometric analysis. Infarct size was measured as the percent of the ischemic area by tetrazolium staining. Myocardial sections were immunohistochemically stained for poly(ADP-ribose) as a measure of poly(ADP ribose) polymerase activity and inhibition. RESULTS: Pigs treated with INO-1001 showed improvements in the +dP/dt at 60 and 90 minutes of post-CPB reperfusion (both p = 0.03) and percent segmental shortening at 30, 60, and 90 minutes of post-CPB reperfusion (p = 0.03, 0.009, and 0.03, respectively). Infarct size was decreased in the treatment group (18.5 +/- 5.7% versus 52.0 +/- 7.7%, INO-1001 versus control, p = 0.03). Poly(ADP-ribose) was reduced in myocardial sections from INO-1001-treated animals compared with controls. CONCLUSIONS: These results suggest that INO-1001 provides myocardial protection by reducing the extent of infarction and improves cardiac function after regional ischemia and cardioplegia CPB. PMID- 12892812 TI - What's new in pediatric surgery. PMID- 12892813 TI - Teaching evidence-based medicine to surgical subspecialty residents. AB - BACKGROUND: Curricula for the teaching of evidence-based medicine to residents have been the subject of reports, analyses, and commentary. Specific programs for teaching evidence-based medicine principles to surgical subspecialty residents have not been identified by the authors. The objective of this article is to report our experience in establishing a teaching program in the principles of evidence-based medicine to surgical subspecialty residents. STUDY DESIGN: We established a teaching program in the principles of evidence-based medicine for neurosurgical residents in a busy neurosurgical training program. Two hours were set aside every other week, replacing traditional professors' rounds with sessions led jointly by a neurosurgeon and an epidemiologist, but based on case presentations from patients currently being treated. From these presentations, searchable clinical questions were developed, and the literature was searched, critically analyzed, and summarized. Results of several cycles on this process are reported. RESULTS: The group developed a repository of Internet-based resources for evidence-based education and practice. Using these resources, the group analyzed six topics in the first 2 years of the program. These included the "best" way to clinically grade patients after subarachnoid hemmorhage, considerations in the biopsy and treatment of enhancing intracerebral mass lesions in patients with acquired immunodeficiency syndrome, the use of prophylactic anticonvulsants in patients with primary brain tumors, the identification of cervical spine injuries in the emergency department, the grading of the fractures of the odontoid process, and the value of removing retained bullets from the spinal canal. The outcomes ranged from finding insufficient evidence to reach a conclusion, through the identification of well conducted and well-reported critical syntheses of the topic already available in literature, to the development of the detailed algorithm for cervical spine clearance that was accepted by the institution's emergency department. CONCLUSIONS: By dedicating some specific time and using resources readily available in most academic health centers, it is possible to incorporate the teaching of the principles of evidence-based practice into the ongoing education of residents on a busy surgical subspecialty service. PMID- 12892815 TI - Treatment of iatrogenic false aneurysms. PMID- 12892816 TI - Should cholecystectomy be performed for asymptomatic cholelithiasis in transplant patients? PMID- 12892817 TI - Winston S Churchill's (1874-1965) inguinal hernia repair by Thomas P Dunhill (1876-1957). PMID- 12892818 TI - Legal considerations for palliative care in surgical practice. PMID- 12892819 TI - Clostridial infection of the abdominal aorta. PMID- 12892820 TI - Epignathus teratoma. PMID- 12892821 TI - Ablative techniques in the treatment of benign and malignant breast disease. PMID- 12892822 TI - Tissue expansion-assisted closure of massive ventral hernias. PMID- 12892823 TI - Restricting fresh frozen plasma in hepatic resections. PMID- 12892825 TI - Enhanced effectiveness of last generation antiblastic compounds vs. cisplatin on malignant pleural mesothelioma cell lines. AB - The purpose of this study was to examine the antiproliferative potentialities of a pool of new generation compounds (Paclitaxel, Docetaxel, Gemcitabine, Topotecan, SN-38) together with fenretinide, a synthetic derivative of retinoic acid, in comparison with the current first choice treatment cisplatin molecule, on a pool of human malignant pleural mesothelioma cell lines derived from either bioptic and pleural effusions samples. To evaluate the chemosensitivity features of malignant mesothelioma cells in vitro, we resorted to a rapid and reproducible colorimetric assay, a useful widely recognized tool for preclinical drug screening. In addition, by DNA content analysis and cellular morphologic assessment, we focused on the apoptosis as a potential mechanism of drug activity. The main results clearly indicate that, in all the models of malignant mesothelioma we handled in vitro, each tested antineoplastic agent is more powerful than cisplatin in inhibiting cell proliferation. Moreover, on experimental evidences basis, we can assume that the cytotoxic activity of tested compounds could be related, at least partially, to the drug-induced programmed cell death. This experimental study gives substance to the expected pharmacologic worth of the second generation antineoplastic drugs even if, in order to afford the most satisfactory biopharmacological approach, allowing to bypass the refractoriness to chemotherapy of this highly lethal tumour, further investigations at preclinical level are required. PMID- 12892826 TI - Effects of U50,488H on transient outward and ultra-rapid delayed rectifier K+ currents in young human atrial myocytes. AB - The effects of trans-(+/-)-3,4-dichloro-N-methyl-N-[2-(1-pyrrolidinyl) cyclohexyl]-benzeneacetamide methanesulfonate salt (U50,488H), a selective kappa opioid receptor agonist, on transient outward K+ current (Ito1) and ultra-rapid delayed rectifier K+ current (IKur) in young human atrial myocytes were evaluated with a whole-cell patch-clamp technique. At +10 mV, U50,488H decreased Ito1 in a concentration-dependent manner (IC50=12.4+/-3.5 microM), while at +50 mV, U50,488H produced biphasic effects on Ito1-increasing and decreasing the current at 1-3 and 10-30 microM, respectively. U50,488H at 10 microM shifted the midpoint (V0.5) of Ito1 activation in a depolarizing direction by approximately 5 mV, accelerated the inactivation, and slowed the recovery from inactivation of Ito1. In addition, U50,488H inhibited IKur in a concentration-dependent manner (IC50=3.3+/-0.6 microM). The effects of U50,488H on the two types of K+ currents were not antagonized by either 5 microM nor-binaltorphimine or 300 nM naloxone. These results indicate that U50,488H affects both Ito1 and IKur in young human atrial myocytes in an opioid receptor-independent manner. PMID- 12892828 TI - Emodin induces apoptosis of human cervical cancer cells through poly(ADP-ribose) polymerase cleavage and activation of caspase-9. AB - Emodin (1,3,8-trihydroxy-6-methylanthraquinone) is an active herbal component traditionally used in China for treating various ailments. Emodin exerts antiproliferative effects in many cancer cell lines and the actual molecular mechanism of which is still not clear. Since apoptosis could be a potential mechanism to explain these effects, we tested whether emodin induces cell death in human cervical cancer cells. Our results suggest that emodin exerts antiproliferative effects in human cervical cancer cells. Emodin inhibited DNA synthesis and induced apoptosis as demonstrated by increased nuclear condensation, annexin binding and DNA fragmentation in Bu 25TK cells in the presence of emodin. Moreover, we demonstrate for the first time in human cervical cancer cells that the apoptotic pathway involved in emodin-induced apoptosis is caspase-dependent and presumably through the mitochondrial pathway, as shown by the activation of caspases-3, -9 and cleavage of poly(ADP-ribose) polymerase. PMID- 12892827 TI - The beta6/alpha5 regions of Galphai2 and GalphaoA increase the promiscuity of Galpha16 but are insufficient for pertussis toxin-catalyzed ADP-ribosylation. AB - Replacement of beta6/alpha5 region at the C-terminus on Galpha16 with Galphaz specific residues has been shown to broaden the promiscuity of Galpha16. Here, we substituted the last 44 residues of Galpha16 with the corresponding region from either Galphai2 or GalphaoA (16i44 and 16o44). 16i44 and 16o44 chimeras were more effective than Galpha16 at coupling to Gi-linked delta-opioid, mu-opioid, and Xenopus melatonin MT1c receptors when coexpressed in green monkey fibroblast (COS 7) cells. 16i44, but not 16o44, also enhanced the formyl peptide-induced stimulation of phospholipase C activity. Both chimeras were resistant to pertussis toxin-catalyzed [32P]ADP-ribosylation, despite the fact that pertussis toxin partially inhibited the chimera-mediated stimulation of phospholipase Cbeta. The use of Galphat1 as a Gbetagamma scavenger revealed that the pertussis toxin-sensitivity can be attributed to endogenous Gbetagamma subunits released from G(i/o). Although incorporation of a Galphai-like beta6/alpha5 region into the C-terminus of Galpha16 increases its promiscuity, this region is not sufficient to support recognition by pertussis toxin. PMID- 12892829 TI - Characterization of binding kinetics of [3H]Tyr-D-Arg2-Phe-Sar4 at opioid receptors. AB - The dermorphin-derived, MOP receptor-selective tetrapeptide Tyr-D-Arg2-Phe-Sar4 (TAPS) exhibits a high antinociceptive potency and stimulates respiration in rats. The receptor binding kinetics of [3H]TAPS were investigated using crude calf thalamic membrane preparations. Saturation studies showed binding of [3H]TAPS at two binding sites (0.4 and 3.2 nM). In the presence of MgSO4, [3H]TAPS binding occurred with high affinity at a single site only. The high affinity binding component was reduced following the addition of K2-EDTA. The increase of the apparent dissociation constant was due to an enhanced dissociation rate (P<0.05), while association rates remained unchanged. Addition of 5'-guanylylimidodiphosphate (Gpp(NH)p) resulted in a reduction of affinity which was augmented in the presence of Na+. Thus, [3H]TAPS, depending on the presence of divalent cations, serves as ligand at two MOP receptor binding sites in the calf thalamus, which may represent distinct affinity states of the same receptor or receptor subtypes thereof. PMID- 12892830 TI - Heme oxygenase is expressed in human pulmonary artery smooth muscle where carbon monoxide has an anti-proliferative role. AB - Heme oxygenase is the rate-limiting enzyme in the catabolism of heme to carbon monoxide, bilirubin and free iron. Many cell types express heme oxygenase-2 constitutively while heme oxygenase-1 is induced at sites of inflammation and oxidative stress. In systemic blood vessels, carbon monoxide may have an important homeostatic role where, like its better-studied counterpart nitric oxide, it is emerging as a vasodilator and an inhibitor of proliferation. However, much less is known regarding the role of heme oxygenase and carbon monoxide in the pulmonary circulation where vascular responses are very different. Here, using primary cultures of human pulmonary artery smooth muscle cells, we present novel data showing that this cell type expresses heme oxygenase 2 constitutively and, in the presence of oxidants, can induce heme oxygenase-1. We also show that the carbon monoxide-releasing molecule, tricarbonyldichlororuthenium (II) dimer, potently and profoundly inhibits proliferation of human pulmonary artery smooth muscle cells. Pulmonary hypertension is a disease characterised by abnormal vascular smooth muscle cell growth and remodelling of the pulmonary vasculature. Our observations support the growing evidence that the heme oxygenase/carbon monoxide system may play a role in the pathology of pulmonary hypertension. PMID- 12892831 TI - Antinociceptive properties of oxymorphazole in the mouse. AB - Oxymorphazole (17-methyl-6,7-dehydro-3,14-dihydroxy-4,5 alpha-epoxy-6,7:3',4' pyrazolomorphinan), a hydrophilic opioid, given intracerebroventricularly (2.5-50 nmol) or intrathecally (0.3-5 nmol) dose-dependently produced tail-flick inhibition in male CD-1 mice. However, oxymorphazole given subcutaneously even at high doses (10-80 mg/kg) produced weak tail-flick inhibition. Oxymorphazole given intraperitoneally (0.1 to 10 mg/kg) dose-dependently inhibited abdominal constriction response induced by intraperitoneally injection of 0.6% acetic acid. Oxymorphazole given intracerebroventricularly (25 nmol) or intrathecally (5 nmol) induced tail-flick inhibition was blocked by pretreatment with the mu-opioid receptor antagonist D-Phe-Cys-Tyr-D-Orn-Thr-Pen-Thr-NH2, but not kappa-opioid receptor antagonist nor-binaltrophimine. The delta-opioid receptor antagonist, naltrindole, blocked the tail-flick inhibition induced by oxymorphazole given intrathecally but not intracerebroventricularly. The inhibition of the abdominal constriction response by oxymorphazole given intraperitoneally was blocked by intraperitoneally pretreatment with naloxone, but not naltrindole or nor binaltrophimine. Thus, oxymorphazole given systemically produces antinociception only with the abdominal constriction test, but not the tail-flick test, suggesting that it produces the antinociception at the peripheral sites when administered systemically. The oxymorphazole-induced antinociception is mainly mediated by the stimulation of mu-opioid receptors when given either centrally or systemically and also the delta-opioid receptors when given intrathecally. The lack of central antinociceptive effect of oxymorphazole given systemically may have interesting clinical implications. PMID- 12892832 TI - Drosophila metabolize 1,4-butanediol into gamma-hydroxybutyric acid in vivo. AB - A solvent, 1,4-butanediol, is also abused as a recreational drug. In mammals, 1,4 butanediol is metabolized into gamma-hydroxybutyric acid (GHB), which stimulates metabotropic gamma-aminobutyric acid (GABA) GABAB and putative GHB receptors. Here we show that in vivo injection of 1,4-butanediol into adult Drosophila leads to GHB synthesis (GHB was detectable 5 min after 1,4-butanediol injection and increased dramatically 1-2 h later). This synthesis of GHB was accompanied by an impairment of locomotor activity that was mimicked by a direct injection of GHB into flies. We propose Drosophila as a model to study the molecular actions of 1,4-butanediol and GHB. PMID- 12892833 TI - Serotonin in the dorsal periaqueductal gray modulates inhibitory avoidance and one-way escape behaviors in the elevated T-maze. AB - The dorsal periaqueductal gray has been implicated in the modulation of escape behavior, a defensive behavior that has been related to panic disorder. Intra dorsal periaqueductal gray injection of serotonin or drugs that mimic its effects inhibits escape induced by electrical or chemical stimulation of this brainstem area. In this study, we investigate whether intra-dorsal periaqueductal gray injection of 5-HT receptor agonists attenuates escape generated by an ethologically based model of anxiety, the elevated T-maze. This test also allows the measurement of inhibitory avoidance, which has been related to generalized anxiety disorder. The effects of the 5-HT receptor agonists were compared in animals with or without a previous exposure to the open arms of the elevated T maze. In these two test conditions, intra-dorsal periaqueductal gray injection of the endogenous agonist serotonin or the 5-HT(2B/2C) receptor agonist m chlorophenylpiperazine (mCPP) enhanced inhibitory avoidance, suggesting an anxiogenic effect. The 5-HT1A receptor agonist 8-hydroxy-2-(di-n-propylamino) tetralin (8-OH-DPAT) impaired this response, suggesting an anxiolytic effect, and the preferential 5-HT2A receptor agonist 2,5-dimethoxy-4-iodoamphetamine (DOI) was ineffective. All these agonists inhibited escape behavior. Apart from mCPP, the effect on escape was detected only in animals pre-exposed to the open arm. None of the drugs tested affected locomotion in the open-field test. Taken altogether, our findings suggest that 5-HT1A and 5-HT2c receptors in the dorsal periaqueductal gray exert opposed control on inhibitory avoidance, implicating these receptors in anxiety conditioning. As previously observed in tests employing the aversive stimulation of the dorsal periaqueductal gray, 5-HT1A and 5-HT2A receptors in this brain area are involved in escape inhibition. Therefore, in different animal models, the activation of these two subtypes of receptors in the dorsal periaqueductal gray consistently attenuates the expression of a panic related behavior. PMID- 12892834 TI - Inhibition of arterial thrombosis by a protease-activated receptor 1 antagonist, FR171113, in the guinea pig. AB - The antiplatelet and antithrombotic effects of FR171113, 3-(4-chlorophenyl)-2 (2,4-dichlorobenzoylimino)-5-(methoxycarbonyl methylene)-1,3-thiazolidin-4-one, a non-peptide protease-activated receptor 1 (PAR1) antagonist, were evaluated in guinea pigs. FR171113 inhibited Ser-Phe-Leu-Leu-Arg-Asn-NH2 (a synthetic PAR1 agonist peptide)-induced and thrombin-induced aggregation of guinea pig platelets in a concentration-dependent manner in vitro (IC50=1.5 and 0.35 microM, respectively). Subcutaneous administration of FR171113 (0.1-3.2 mg/kg) produced a dose-dependent inhibition of platelet aggregation ex vivo. The ED50 value of FR171113 for platelet aggregation was 0.49 mg/kg s.c. However, FR171113 did not have an inhibitory effect on ADP- or collagen-induced platelet aggregation in vitro and ex vivo. One hour after FR171113 treatment at 1.0 mg/kg s.c., significant inhibition of arterial thrombosis without a prolongation of thrombin time or coagulation time was seen in the FeCl3-induced carotid artery thrombosis model in guinea pigs. Furthermore, FR171113 did not prolong bleeding time even at 32 mg/kg s.c., which is a much higher dose than that required in the thrombosis model. These observations indicate that FR171113 has desirable antiplatelet effects both in vitro and in vivo and that its in vivo antithrombotic activity is efficacious without causing a prolongation of bleeding time. PMID- 12892835 TI - Therapeutic effects of ghrelin on endotoxic shock in rats. AB - We investigated the effects of ghrelin in a rat endotoxic shock model, and also observed the direct role of endotoxin on ghrelin generation in gastric mucosa. About 55% (11/20) of rats treated with lipopolysaccharide (5 mg/kg i.v.) alone died within 24 h of endotoxin injection. However, administration of ghrelin either at the same time as lipopolysaccharide injection (early treatment) or 12 h after lipopolysaccharide injection (late treatment) significantly decreased the mortality rate and ameliorated the hypotension seen in rats with endotoxic shock. Early and late treatment with ghrelin increased markedly the plasma glucose concentration and decreased the plasma lactate concentration. Early treatment with ghrelin attenuated significantly the deficiency in myocardial ATP content, but late treatment with ghrelin had no effect on myocardial ATP content. The plasma ghrelin level was significantly increased in the rats with endotoxin shock, and it increased further after ghrelin administration. Exposure of rat gastric mucosa in vitro to lipopolysaccharide (1.0 to 100 microg/ml) triggered the release of ghrelin from mucosa tissue in a dose- and time-dependent manner, meaning that lipopolysaccharide stimulated directly gastric mucosa to synthesize and secrete ghrelin. The results suggest that ghrelin could have therapeutic value for endotoxic shock. PMID- 12892836 TI - Rat liver ischemia-reperfusion-induced apoptosis and necrosis are decreased by FK506 pretreatment. AB - The aim of this study was to demonstrate that tacrolimus (FK506) has a hepatoprotective effect by reducing ischemia-reperfusion-induced apoptosis and necrosis, both of which lead to post-surgical liver dysfunction. An ischemia reperfusion model and primary cultured rat hepatocytes subjected to hypoxic and reoxygenation phases, mimicking the surgical process, were used. c-Jun N-terminal kinase 1/stress-activated protein kinase 1 (JNK1/SAPK1) activation leads to caspase 3 activation, a trigger of apoptosis. The activation status of JNK1/SAPK1 was evaluated by immunoprecipitation or Western-blotting experiments. Apoptosis was assessed by measuring caspase activation and by TUNEL (terminal deoxynucleotidyltransferase-mediated deoxyuridine triphosphate-biotin nick-end labeling) reaction. Necrosis was assessed histologically. Tacrolimus improved the survival rate of rats subjected to ischemia-reperfusion. After FK506 pretreatment, the liver necrosis rate was reduced, and ischemia-reperfusion induced JNK1/SAPK1 activation and apoptosis were significantly decreased. In hypoxia-reoxygenation-subjected hepatocytes, tacrolimus reduced JNK1/SAPK1 and caspase 3 activation. In the liver, tacrolimus prevented ischemia-reperfusion induced apoptosis and necrosis. PMID- 12892837 TI - Dopamine D1 receptor-dependent inhibition of NaCl transport in the rat thick ascending limb: mechanism of action. AB - Our previous in vitro microperfusion studies established that dopamine inhibits sodium chloride transport in the rat medullary thick ascending limb. The present study was designed to determine the intracellular signaling pathway mediating this response. The dopamine D1 receptor agonist fenoldopam (1 microM) inhibited sodium chloride transport in the thick ascending limb by 42+/-5%. The dopamine D1 receptor antagonist R-(+)-7-Chloro-8-hydroxy-3-methyl-1-phenyl-2,3,4,5-tetrahydro 1H-3-benzazepine-HCl (SCH-23390) completely blocked this effect of fenoldopam. Suppression of protein kinase A activity using either myristoylated protein kinase inhibitor (PKI) or N-[2-(p-Bromocinnamylamino)ethyl]-5 isoquinolinesulfonamide.2HCl (H-89), as well as suppression of phospholipase C activity using 1-(6-((17 beta-3-methoxyestra-1,3,5(10)-trien-17-yl)amino)hexyl) 1H-pyrrole-2,5-dione (U-73122), had no effect on fenoldopam-dependent inhibition of transport. In contrast, inhibition of phospholipase A2 activity using E-6 (Bromomethylene)tetrahydro-3-(1-naphthalenyl)-2H-pyran-2-one (HELSS) significantly attenuated the effect of fenoldopam by 74%. The cytochrome P-450 monooxygenase inhibitor 17-octadecynoic acid (17-ODYA) and the protein kinase C inhibitor staurosporine both significantly attenuated the effects of fenoldopam by 67%. Exposure to 20-Hydroxy-(5Z, 8Z, 11Z, 14Z)-eicosatetraenoic acid (20-HETE) inhibited transport by 31+/-5%, and this effect was significantly attenuated by 66% in the presence of staurosporine. We propose a signaling pathway in which dopamine activates a calcium-independent phospholipase A2 in the medullary thick ascending limb. Released arachidonic acid is then metabolized to 20-HETE which subsequently increases protein kinase C activity that acts as a final transport effector. PMID- 12892838 TI - Enhancement of natural killer cell function by titanocenes in mice bearing Ehrlich ascites tumour. AB - In the present work, we studied the effects of two titanocenes, biscyclopentadienyldichlorotitanium IV, (DDCT) and its derivative, biscyclopentadienylditiocianatetitanium IV (BCDT), on the activity of natural killer (NK) cells in Ehrlich ascites tumour (EAT)-bearing BALB/c mice. In order to investigate a more direct effect of these compounds on NK cell function, we performed experiments with severe combined immunodeficiency (SCID) mice, which exhibit a normal NK cell response in the absence of T and B cells. The treatment consisted of intraperitoneal (i.p.) administration of 15 mg/kg/day of DDCT for 2 days or 10 mg/kg/day of BCDT for 3 days. In addition, to verify whether the effects produced by the titanocenes were compound specific or related to a direct antitumour effect, we also investigated the effects of a 3-day treatment with 100 mg/kg of cyclophosphamide cyclophosphamide on NK cell activity. Our results demonstrated that, in BALB/c and SCID mice, NK cell function declined to subnormal levels after inoculation of the tumour. In these animals, although treatment with DDCT and BCDT significantly enhanced NK cell function, only DDCT restored NK cell activity to normal values in all stages studied. Conversely, treatment with cyclophosphamide reduced NK cell function in nontumour bearing SCID mice and was also unable to restore the decreased NK activity of tumour bearing SCID mice, thus demonstrating that the enhancement of NK cell function by titanocenes is compound specific. The same effect of cyclophosphamide was observed with BALB/c mice. In the present study, the up-modulatory effects of these two compounds on NK cell function reveal a new aspect of the mechanism of antitumoural action of titanocenes. PMID- 12892839 TI - Interleukin-1beta, interleukin-6, tumour necrosis factor-alpha and interferon gamma released by a viral infection and an aseptic inflammation reduce CYP1A1, 1A2 and 3A6 expression in rabbit hepatocytes. AB - Inflammation reduces activity and expression of hepatic cytochrome P450 (P450) and therefore diminishes drug biotransformation. This study aimed to identify the serum mediators triggered by a viral infection and an aseptic inflammation that downregulate P450 isoforms. Incubation of hepatocytes with serum from rabbits with a turpentine-induced inflammation or humans with a viral infection decreased the amount of cytochrome 1A1 (CYP1A1), 1A2 and 3A6 mRNA and apoproteins. By serum fractionation and immuno-neutralization, we showed that in the aseptic inflammation, interleukin-6 and, to a lesser degree, interleukin-1beta are involved in the downregulation of all three isoforms. In serum from humans with a viral infection, interleukin-1beta, interleukin-6, interferon-gamma and tumour necrosis factor-alpha contribute to the downregulation of P450 isoforms. CYP1A1 and 1A2 are regulated by serum mediators at the transcriptional level, while the expression of CYP3A6 appears to be under the control of pre- and posttranscriptional mechanisms. PMID- 12892840 TI - Ischemic preconditioning reduces the severity of ischemia/reperfusion-induced pancreatitis. AB - In various organs, including heart, kidneys, brain, liver and stomach, preconditioning by brief exposure to ischemia protects the organ against damage evoked by subsequent severe ischemia. This study has been undertaken to check whether two brief ischemic periods protect the pancreas against severe ischemia/reperfusion-induced pancreatitis and, if so, what is the role of sensory and vagal nerves in this phenomenon. In male Wistar rats, the ischemic preconditioning of the pancreas was performed by clamping of celiac artery (2 x 5 min with 5 min interval). Thirty minutes after preconditioning or sham operation, the ischemia/reperfusion-induced pancreatitis was evoked by clamping of inferior splenic artery for 30 min using microvascular clips, followed by 1 h reperfusion. Sensory nerves ablation was induced 10 days before final experiments by capsaicin. Truncal vagotomy was performed 1 week before the experiment. Exposure to regular 30-min pancreatic ischemia followed by 1 h reperfusion led to the development of acute hemorrhagic pancreatitis. Ischemic preconditioning, applied prior to induction of pancreatitis, caused the reduction in plasma lipase, plasma interleukin-1beta and histological signs of pancreatic damage, as well as attenuated the reduction in pancreatic blood flow and DNA synthesis. Ablation of sensory nerves by capsaicin caused an aggravation of ischemia/reperfusion-induced pancreatic damage and attenuated a protective effect of ischemic preconditioning. Noxious effect of sensory nerves ablation on the pancreas was accompanied by the reduction in pancreatic blood flow and an increase in plasma interleukin-1beta. Similar but less pronounced deleterious effect on the pancreas was observed after vagotomy. We conclude that: (1) pancreatic ischemic preconditioning reduces the severity of ischemia/reperfusion-induced pancreatitis; (2) this effect seems to be related, at least in part, to the improvement of pancreatic blood flow and the reduction in the release of proinflammatory interleukin-1beta; (3) sensory and vagal nerves are involved in protective effect of ischemic preconditioning against pancreatic damage. PMID- 12892841 TI - Extrahypothalamic corticotropin-releasing hormone mediates (-)-nicotine-induced elevation of plasma corticosterone in rats. AB - (-)-Nicotine activates the hypothalamic-pituitary-adrenal axis via an activation of the brainstem catecholaminergic neurons in rats. The present study was undertaken to clarify the mechanisms involved in the (-)-nicotine-induced activation of brainstem catecholaminergic neurons in anesthetized rats. Physostigmine (a cholinesterase inhibitor) (0.31 and 0.77 micromol/animal, i.p.) dose-dependently elevated plasma corticosterone in the presence of scopolamine (a muscarinic receptor antagonist) (2.3 micromol/animal, i.p.). (-)-Nicotine (250 and 500 nmol/animal, i.c.v.) dose-dependently elevated plasma corticosterone with concomitant noradrenaline release in the hypothalamic paraventricular nucleus. The (-)-nicotine (500 nmol/animal, i.c.v.)-induced elevation of corticosterone was abolished by phentolamine (an alpha-adrenoceptor antagonist) (0.66 micromol/animal, i.c.v.), and attenuated by (+/-)-sotalol (a beta-adrenoceptor antagonist) (0.97 micromol/animal, i.c.v.). The (-)-nicotine-induced increases of plasma corticosterone and hypothalamic noradrenaline release were abolished either by hexamethonium (a nicotinic acetylcholine receptor antagonist) (1.8 micromol/animal, i.c.v.), CP-154,526 (butyl-ethyl-[2,5-dimethyl-7-(2,4,6 trimethylphenyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl]amine) (a selective CRF-1 receptor antagonist) (1.3 micromol/animal, i.c.v.) or indomethacin (a cyclooxygenase inhibitor) (1.2 micromol/animal, i.c.v.). These results suggest that (-)-nicotine elevates plasma corticosterone by CRF-1 receptor- and prostaglandin-mediated noradrenaline release in the paraventricular nucleus in rats. PMID- 12892842 TI - Ethanol rapidly induces steroidogenic acute regulatory protein expression and translocation in rat adrenal gland. AB - Acute ethanol exposure increases GABAergic neuroactive steroids in plasma and brain by releasing these steroids or their precursors from the adrenal glands. The present study showed that ethanol administration rapidly increases the expression of steroidogenic acute regulatory protein (StAR) in the cytosolic and mitochondrial fractions of adrenal glands. The increased StAR protein expression paralleled increases in plasma pregnenolone, progesterone and corticosterone levels. The rapid synthesis and translocation of StAR protein in adrenals likely represent the mechanism of ethanol-induced increases in neuroactive steroids. PMID- 12892843 TI - EMD 281014, a new selective serotonin 5-HT2A receptor antagonist. AB - The 5-HT2A receptor ligand 7-[4-[2-(4-fluoro-phenyl)-ethyl]-piperazine-1 carbonyl]-1H-indole-3-carbonitrile HCl (EMD 281014) selectively binds to human (h) and rat 5-HT2A receptors (IC50 values 0.35 and 1 nM, respectively; vs. 1334 nM for h5-HT2C) and inhibited 5-HT-stimulated [35S]guanosine 5'-O-3 thiotriphosphate (GTPgammaS)-accumulation in h5-HT2A transfected Chinese hamster ovary cells (IC50 9.3 nM). EMD 28014 counteracted the N-ethoxycarbonyl-2-ethoxy 1,2-dihydroquinoline (EEDQ)-induced decrease of [3H]ketanserin binding in rat frontal cortex (ID50 0.4 mg/kg p.o.) and R-(-)-1-(2,5-dimethoxy-4-iodophenyl) aminopropane (DOI)-induced head-twitch behaviour in mice (ID50 0.01 mg/kg s.c., 0.06 mg/kg p.o.), demonstrating unique selectivity and efficacy. PMID- 12892844 TI - Universal stress proteins and Mycobacterium tuberculosis. AB - Mycobacterium tuberculosis expresses universal stress proteins (USPs) when its growth is retarded by oxygen depletion. This class of proteins is emerging as being important in the resistance of bacteria to stress and prolonged growth arrest. Here we assess the properties of USPs and their relevance to mycobacteria. PMID- 12892845 TI - Soil microbial activities and carbon and nitrogen fixation. AB - Soil microbial activity that reflects microbiological processes of soil microorganisms is the potential indicator of soil quality, as plants rely on soil microorganisms to mineralize organic nutrients for growth and development. Soil microorganisms also process plant litter and residues into soil organic matter, a direct and stable reservoir of carbon and nitrogen that consists of living and dead organic materials subject to rapid biological decomposition. In natural systems, the action of soil microorganisms is a major determinant of efficient nutrient cycling. This paper reviews the current state of knowledge on the fate of soil microorganisms in terms of carbon and nitrogen fixation. PMID- 12892846 TI - Oxalotrophic bacteria. AB - Oxalic acid and its salts are widespread in nature, as they are produced by many species of plants, algae and fungi. The bacteria, which are capable of using oxalate as a sole carbon and energy source, are described as being "oxalotrophic". Oxalotrophic bacteria do not constitute a homogeneous taxonomic group, but they do constitute a well-defined physiological group. A limited number of aerobic bacteria which are able to utilize oxalate as sole carbon and energy source have been completely described. Most of them are facultative methylotrophs and/or facultative hydrogen-oxidizing chemolithoautotrophs. In this review, the current status of the taxonomy and biodiversity of oxalotrophic bacteria in various environments, and aspects of their biotechnological potential, are briefly summarized. PMID- 12892847 TI - Metal tolerance and biosorption capacity of Bacillus circulans strain EB1. AB - A heavy-metal-resistant bacterium Bacillus sp., strain EB1 was isolated from heavy-metal-contaminated soil in the southeast region of Turkey. Based on 16S ribosomal DNA sequencing, the microorganism was closely related to Bacillus circulans. Minimal inhibitory concentrations of metals (MICs) for the bacterium were determined. Bacillus EB1 exhibited high MIC values for metals and a large spectrum of antibiotic resistance. The order of toxicity of the metals to the bacterium was Cd=Co>Cu>Ni>Zn>Mn in solid media. The effects of increasing metal concentrations to the growth rate were determined in order to obtain precise patterns of resistance in liquid cultures. From the results of heavy metal toxicity, inhibitory concentrations in solid media were higher than those in liquid media. Metal biosorption was determined during the course of growth. B. circulans strain EB1 was capable of removing 90% of Mn, 68% of Zn, 65% of Cu, 45% of Ni and 40% of Co during the active growth cycle with a specific biosorption capacity of 25, 22, 20, 13 and 12 mg/l, respectively. Since Bacillus cells could grow in the presence of significant concentrations of metals and due to high metal biosorption capacity in aerobic conditions, this bacterium may be potentially applicable in in situ bioremediation of heavy-metal-contaminated aqueous systems. PMID- 12892848 TI - Envelope glycosylation determined by lectins in microscopy sections of Acinetobacter venetianus induced by diesel fuel. AB - It was suggested in a previous study that cells of Acinetobacter venetianus VE-C3 adhere to diesel fuel by synthesizing a capsular polysaccharide containing glucose and/or mannose. To study the fine structure of cells and localization of bacterial polysaccharide in the presence of diesel fuel, two lectins were used: ConA, an agglutinin from Canavalia ensiformis specific for mannose and/or glucose residues, and PNA, an agglutinin from Arachis hypogaea, for terminal galactose residues. The lectins were conjugated with electron dense ferritin for transmission electron microscopy (TEM) and with fluorescein isothiocyanate (FITC) for scanning confocal laser microscopy (SCLM). Samples were prepared by freeze substitution, which allows glycosylation to be determined in situ in thin sections of specimens. The distribution of glycosylation was imaged with and without treatment of specimens with their specific hapten (glucose and galactose). The glycosylation activity produced a polysaccharide capsule. Emulsified diesel fuel nanodroplets were observed at the cell envelope perimeter. Fine structure of vesicles consisted of polysaccharide and diesel fuel nanodroplets. Lectin blotting analysis showed ConA-positive glycoprotein with an apparent molecular mass of 22 kDa in the outer membrane. Its production was induced by diesel fuel. This glycoprotein was probably responsible for bioemulsifying activity at the cell envelope. Several other glycoproteins were positive for PNA lectin, the main constituent migrating with an apparent molecular weight of 17.8 kDa. However, they were all constitutive and probably involved in cell biofilm formation at the oil surface. PMID- 12892849 TI - Genetic engineering of Enterobacter aerogenes with the Vitreoscilla hemoglobin gene: cell growth, survival, and antioxidant enzyme status under oxidative stress. AB - Hemoglobins in unicellular organisms, like the one here in the bacterium Vitreoscilla, have greater chemical reactivity than their homologues in multicellular organisms. They can catalyze redox reactions and may protect cells against oxidative stress. The ability of Vitreoscilla hemoglobin to complement deficiencies of terminal cytochrome oxidases in Escherichia coli also suggests that this hemoglobin can receive electrons during respiration. In this study, a recombinant strain of Enterobacter aerogenes engineered to produce the Vitreoscilla Hb was investigated with regard to its susceptibility to oxidative stress. The culture response to oxidative stress produced by exogenously applied hydrogen peroxide was characterized in terms of cell growth, survival and the activities of two key antioxidant enzymes (catalase and superoxide dismutase). The influence of the physiological state of the cells and different media upon these culture dynamics was determined. Results showed that the hemoglobin expressing strain is quite distinct in terms of growth/survival properties and activity of antioxidant enzymes from that of non-hemoglobin counterparts. PMID- 12892850 TI - Exopolysaccharide synthesis in Rhizobium leguminosarum bv. trifolii is related to various metabolic pathways. AB - Rhizobium leguminosarum bv. trifolii synthesizes extracellular polysaccharide (EPS) that is postulated to be a biologically active signalling molecule in clover symbiosis. A group of seven exopolysaccharide-deficient (Exo), non nitrogen-fixing mutants of R. leguminosarum bv. trifolii strain 24.1 isolated by transposon mutagenesis were complemented to mucoid phenotype by a low-copy plasmid carrying the pssA gene encoding the first glucosyl-IP-transferase. Some of these mutants were not corrected in their symbiotic defect by the pssA gene. Precise localization of Tn5 insertion sites by subcloning and sequencing the adjacent genomic DNA in the Exo mutants identified the disrupted genes and their possible functions. Only one mutant (Rt74) was mutated in pssA gene; others were mutated in diverse genes that were not directly involved in EPS biosynthesis. The suppression of EPS deficiency in these mutants by additional copies of pssA indicated a possible connection between exopolysaccharide biosynthesis and various metabolic pathways. PMID- 12892851 TI - Vir90, a virulence-activated gene coding for a Bordetella pertussis iron regulated outer membrane protein. AB - Bordetella pertussis undergoes phenotypic changes modulated by the bvgAS locus, which regulates the expression of many genes related to virulence and immunogenicity. We previously reported the N-terminal sequence of a 90 kDa bvg regulated outer membrane protein (OMP) of B. pertussis (SWISS-PROT accession No. p81549), a novel potential virulence factor that we named Vir90. The open reading frames (ORFs) which potentially code for Vir90 in B. pertussis, B. parapertussis and B. bronchiseptica were identified by computer analysis of the genomic sequences available for the three Bordetella species. Nucleotide sequence analysis of the vir90 upstream region revealed the presence of a putative promoter, a BvgA binding site and a putative Fur binding site. The B. pertussis Vir90 protein showed significant homology with ferrisiderophore receptors from Gram-negative bacteria. An antiserum raised against Vir90His recombinant protein recognized the 90-kDa protein in immunoblots of OMPs from these three virulent Bordetella species. The accumulation of the Vir90 protein increased 4-fold under low iron growth conditions. Therefore, the vir90 gene is expressed in the tested species and its expression is regulated positively by the BvgAS system and negatively under high iron concentration, likely by Fur. PMID- 12892852 TI - A new mini-transposon for in vivo protein epitope tagging: application to Burkholderia multivorans. AB - A short amino acid sequence coding for the mature Pseudomonas aeruginosa OprI lipoprotein was fused to a mini-Tn5 plasposon (mini-transposon with an origin of replication) with tetracycline resistance in order to generate in-frame fusion proteins after transposition. After conjugative transfer to Burkholderia multivorans, clones reacting with an anti-OprI mab were selected. In-frame OprI tagged proteins were detected and identified for six clones. The six C-tagged proteins were detected by immunoblot. The different mutants had insertions into a histone H1-like coding gene, cspD, encoding a cold-shock protein, dsbC, encoding a putative outer membrane lipoprotein involved in thiol-disulfide exchange, paaE, a ferredoxin-NADPH reductase gene, a gene for the catabolism of propionate, and one encoding an unknown protein. PMID- 12892853 TI - Cognitive therapy for schizophrenia: a preliminary randomized controlled trial. AB - BACKGROUND: The aim of the current study was to assess whether patients with a DSM-IV diagnosis of schizophrenia and experiencing persistent positive and negative symptoms improve with the addition of cognitive-behavioural therapy to enriched standard treatment. METHODS: A controlled study was completed with 42 patients randomized to either cognitive-behavioural therapy plus enriched treatment-as-usual (CBT-ETAU) (n = 24) or enriched treatment-as-usual only (ETAU) (n = 18). Enriched treatment-as-usual comprised comprehensive treatment within specialised schizophrenia treatment services. Cognitive-behavioural therapy was conducted on an individual basis for 6 months (20 sessions). Clinical assessments were done at pretreatment, posttreatment and at 6-month follow-up by raters blind to group allocation. RESULTS: Significant clinical effects were observed for positive, negative and overall symptom severity for patients treated in CBT-ETAU, although there were no statistically significant differences between the treatment groups at posttreatment. The most pronounced effect of CBT-ETAU in comparison to ETAU in this study was in the reduction of negative symptoms at follow-up. CONCLUSION: These results show promise for the impact of CBT on negative symptoms when explicitly targeted in treatment. PMID- 12892854 TI - Antisaccade performance is abnormal in schizophrenia patients but not in their biological relatives. AB - Numerous studies have replicated the finding that schizophrenia patients make an increased number of errors on an antisaccade task. Some studies have reported that relatives of schizophrenia patients also make an increased number of antisaccade errors, a finding that has been interpreted to support the usefulness of compromised antisaccade performance as an index of genetic liability for schizophrenia. We examined performance on an antisaccade task in schizophrenia patients, nonpsychiatric controls, first-degree relatives of schizophrenia patients and first-degree relatives of nonpsychiatric controls. Schizophrenia patients made significantly more errors than did nonpsychiatric controls, but relatives of schizophrenia patients did not differ from relatives of controls or from all controls. Increased antisaccade errors on the standard version of the antisaccade task are associated with schizophrenia, but do not seem to be a co familial trait for schizophrenia. PMID- 12892855 TI - Sensitivity and specificity of select biological indices in characterizing psychotic patients and their relatives. AB - BACKGROUND: Although studies have detailed biological abnormalities in schizophrenia patients and their first-degree biological relatives, few studies have directly compared the utility of biological indices in these individuals. METHODS: Measures of global smooth-pursuit ocular motor (OM) function, low frequency and alpha band electroencephalogram (EEG) power, and nonspecific fluctuations (NSF) in electrodermal activity and visibility of the plexus in the nailfold were collected from 136 schizophrenia patients and 67 of their first degree biological relatives, 71 affective disorder psychotic patients and 68 of their first-degree biological relatives, and 169 nonpsychiatric comparison subjects. We conducted receiver operator characteristic (ROC) analyses to determine how well each index differentiated the patient groups and the groups of first-degree relatives. RESULTS: Smooth-pursuit ocular motor function, low frequency and alpha band EEG power, and nailfold plexus visibility differentiated schizophrenia patients from nonpsychiatric comparison subjects. Nailfold plexus visibility was the only measure that significantly differentiated schizophrenia patients from both nonpsychiatric controls and affective patients. Smooth-pursuit ocular motor function and the number of electrodermal nonspecific fluctuations differentiated relatives of schizophrenia patients from nonpsychiatric comparison subjects. CONCLUSION: Increased nailfold plexus visibility may mark a process associated with abnormal brain development leading to schizophrenia. Smooth pursuit dysfunction may mark genetic vulnerability that is relatively specific to schizophrenia. PMID- 12892856 TI - Components of the smooth pursuit function in deficit and nondeficit schizophrenia. AB - The diagnosis of schizophrenia likely encompasses a heterogeneous group of disorders, complicating the search for its causes. Studies of deficit schizophrenia represent an attempt to reduce this heterogeneity by identifying biologically distinct subgroups. Supplementing clinical phenotypes with biological markers of risk (e.g., eye-tracking and sensory-gating deficits) have also been used to reduce disease heterogeneity. In this study, we examined smooth pursuit eye movements in healthy controls (n = 37), and deficit (n = 18) and nondeficit (n = 32) patients with schizophrenia to determine what aspects of abnormal smooth pursuit are associated with the two patient groups, and which, if any, specifically mark the deficit phenotype. A small sample of relatives of deficit (n = 12) and nondeficit (n = 35) patients was also examined. Positive symptoms were equally present in deficit and nondeficit patients. Subtle, psychotic-like positive traits were also equally present in the relatives of both deficit and nondeficit probands, whereas negative symptoms were significantly more prevalent among the relatives of deficit probands. Deficits in predictive pursuit were present in both patient groups and both groups of relatives. Deficit patients showed significantly lower initiation acceleration. A similar pattern of results was seen in our pilot sample of relatives of deficit patients. These findings suggest that predictive smooth pursuit abnormality is associated with positive symptoms in schizophrenia, and that initiation abnormalities may be associated with the deficit syndrome. PMID- 12892857 TI - Cognitive predictors of medication adherence among middle-aged and older outpatients with schizophrenia. AB - OBJECTIVE: Medication nonadherence presents a considerable problem in patients with schizophrenia. There are limited and conflicting data on the association of cognitive impairment with antipsychotic nonadherence. In this study, we evaluated the correlation of patients' scores on Mattis' Dementia Rating Scale (DRS; total and subscale scores) with scores on the Medication Management Ability Assessment (MMAA), a performance-based measure of medication management. METHODS: Participants included 110 outpatients with schizophrenia or schizoaffective disorder. Each was evaluated using the MMAA role-play tasks and the DRS. Patients also completed the Drug Attitude Inventory (DAI), and the PANSS (Positive And Negative Syndrome Scale). RESULTS: Age, DAI score, and DRS scores were all correlated with MMAA performance. In a stepwise regression analysis, only DRS scores were predictive of MMAA performance. Among the DRS subscales, conceptualization and memory were the best statistical predictors of MMAA performance. CONCLUSION: Cognitive functions, especially conceptualization and memory, were the strongest patient-related predictors of his or her ability to manage medications, over and above the effects of age, gender, education level, symptom severity, and attitudes toward medications. These results suggest a need for intervention studies focused on improving, or at least compensating for, specific cognitive deficits such as those in memory and conceptualization among patients with schizophrenia in order to improve their ability to manage medications. PMID- 12892858 TI - Differences in smooth pursuit eye movement between posttraumatic stress disorder with secondary psychotic symptoms and schizophrenia. AB - Studies show high comorbidity between post-traumatic stress disorder and psychotic symptoms. Despite this fact, there has been only one published study of the neurobiology of this enigmatic disorder. This preliminary study examines the relationship between psychotic symptoms in post-traumatic stress disorder (PTSD) and schizophrenia by measuring smooth pursuit eye movement (SPEM) in subjects with PTSD and secondary psychotic symptoms, schizophrenia, and healthy controls. The results show that PTSD with secondary psychotic symptoms is associated with a SPEM deficit that is different from the SPEM deficit associated with schizophrenia. PMID- 12892859 TI - Source distribution of neuromagnetic slow-wave activity in schizophrenic patients -effects of activation. AB - When slow waves in the EEG delta and theta frequency range appear in the waking state, they may indicate pathological conditions including psychopathology. The generators of focal slow waves can be mapped using magnetic source imaging. The resulting brain maps may possibly characterize dysfunctional brain areas. The present study examined the stability of the density and distribution of MEG slow waves during three conditions-rest, mental arithmetic and imagery-in 30 schizophrenic patients and 17 healthy controls. Schizophrenic patients displayed a higher density of delta and theta generators primarily in temporal and parietal areas. The group difference was not affected by the particular conditions. The focal concentration of delta and theta slow waves did not differ between patients with and without neuroleptic medication, whereas the prominence of theta dipoles in the temporal area correlated with neuroleptic dosage. The relative amount of temporal slow waves was correlated with the negative symptoms score (PANSS-N) suggesting that temporal dysfunction may be related to negative symptomatology.Results suggest that the distribution of slow-wave activity, measured in a standardized setting, might add diagnostic information about brain abnormalities in schizophrenia. PMID- 12892861 TI - Apathy in schizophrenia: clinical correlates and association with functional outcome. AB - Apathy is considered one of the negative symptoms of schizophrenia, but its natural history and relationship to other clinical characteristics have not been systematically studied. The purpose of this cross-sectional study was to measure the level of apathy in schizophrenia and its relation to other symptoms and functional outcome. Twenty-eight patients with schizophrenia, and receiving antipsychotic treatment, were assessed with the Apathy Evaluation Scale (AES). The mean level of apathy of patients with schizophrenia, as rated by the AES, was significantly higher than that of matched healthy control subjects. In the patients, apathy was not significantly correlated with positive symptoms or depressive symptoms. It was significantly correlated with the item "emotional withdrawal" on the negative subscale of the Positive and Negative Syndrome Scale (PANSS), but was not correlated with the overall negative subscale score. Apathy was more highly associated with functional outcome than were other symptom measures, and it was independently associated with functional outcome above and beyond other negative symptoms. It was not associated with observed interest in playing a video game or performance on a simulated clerical task. PMID- 12892860 TI - Low maternal vitamin D as a risk factor for schizophrenia: a pilot study using banked sera. AB - OBJECTIVE: Evidence from epidemiology suggests that low maternal vitamin D may be a risk factor for schizophrenia. METHOD: Based on sera taken during the third trimester, we compared the level of 25 hydroxyvitamin D3 in mothers of individuals with schizophrenia or schizoaffective disorders versus mothers of unaffected controls. For each case, we selected two controls matched on race, gender and date of birth of the offspring. RESULTS: There was no significant difference in third trimester maternal vitamin D in the entire sample (cases = 26, controls = 51). Within the subgroup of black individuals (n = 21), there was a trend level difference in the predicted direction. CONCLUSIONS: Maternal vitamin D does not operate as a continuous graded risk factor for schizophrenia, however, the results in the black subgroup raise the possibility that below a certain critical threshold, low levels of maternal vitamin D may be associated with an increased risk of schizophrenia. PMID- 12892862 TI - Sex differences in symptoms of psychosis in a non-selected, general population sample. AB - BACKGROUND: Little is known about sex differences in psychosis beyond the borders of clinical disorder. METHODS: A general population sample of 7,076 subjects was assessed using the Composite International Diagnostic Interview, in order to explore sex differences in the prevalence of any positive and negative symptoms of psychosis, and to examine to what degree any differences could be explained by differences in level of affective symptoms. RESULTS: Male sex was associated with higher prevalence of negative symptoms (OR = 1.6, 95% CI = 1.0, 2.5), independent of differences in affective symptoms and presence of DSM-III-R psychotic disorder. Women had higher rates of positive psychotic experiences (OR = 0.8, 95% CI = 0.7, 0.9), but this difference disappeared after adjustment for depressive symptoms (adjusted OR = 1.2, 95% CI = 0.9, 1.5). CONCLUSION: The sex differences in psychopathology that are seen in schizophrenia are expressed beyond the clinical phenotype, suggesting sex-dependent continuous and normal variation of several psychosis dimensions. The higher rates of positive psychotic experiences seen in women may be secondary to differences in the rate of affective symptoms. PMID- 12892863 TI - The effects of atypical antipsychotic medications on psychosocial outcomes. AB - This review examines the question--Do atypical antipsychotic medications improve psychosocial outcomes?--by examining studies that compared the effects of atypicals to conventional antipsychotics or to other atypical medications. The authors reviewed randomized clinical trials of atypical antipsychotic medication that included psychosocial variables as outcomes. Findings from 31 published studies on more than 12,000 individuals generally showed that atypical medications led to significant improvements in negative symptoms compared to conventional antipsychotics. Effects on global assessment of psychosocial functioning and on the quality of life were mixed with only about half the studies reporting significant improvements. Olanzapine yielded the best results on psychosocial functioning; remoxipride was found to yield few significant changes on these variables. This review provides evidence that some atypical antipsychotics may have direct effects on some of the psychosocial disabilities that result from serious mental illness. PMID- 12892864 TI - A comparison between an interview and a self-report method of insight assessment in chronic schizophrenia. AB - We examined the relationship between a researcher-rated and a self-report method of insight assessment in patients with schizophrenia. Previous research has established that a moderate correlation exists between several researcher-rated and self-report insight scales. Hence, we wanted to investigate this association using another combination of assessment techniques. Accordingly, 34 patients with chronic schizophrenia were administered the Scale to Assess Unawareness of Mental Disorder (SUMD) and the Birchwood self-report Insight Scale (IS). We found a significantly lower report of insight on the Birchwood IS, regardless of administration order. These findings suggest that inherent bias within the patient-examiner interaction may be responsible for this discrepancy. Overall, a significant positive correlation was observed between total scores on the SUMD and the Birchwood self-report IS. Unexpectedly, there was a remarkable difference in correlation with respect to the order of test administration. The "self-report first" group's scores were strongly correlated, whereas the "researcher-rated first" group's scores showed little relation. These results demonstrate the importance of investigating the effects of administration order, and also question the reliability of other correlational studies between researcher-rated and self-report insight scales that have failed to employ a counterbalanced research design. PMID- 12892865 TI - Future employability, a new approach to cost-effectiveness analysis of antipsychotic therapy. AB - OBJECTIVE: Schizophrenia patients pose a substantial burden in terms of indirect costs, much of which is attributable to loss of employment. We present a new approach to assess the cost-effectiveness (CE) of risperidone vs. haloperidol using employability as an outcome measure. METHODS: A decision analytic CE model was developed to compare the two treatments over a 1-year period including all direct medical costs and the number of employable persons as a measure of effectiveness. A measure of executive functioning, the Wisconsin Card Sorting Test Category score (WCST-Cat score), was used as an intermediate endpoint from which employability was modeled. A Monte Carlo procedure, using WCST-Cat score sampling distributions from clinical trials, simulated the WCST-Cat score distribution for a cohort of 10,000 patients. A clinically stable patient, with a Positive and Negative Syndrome Scale (PANSS) score increase of at least 20% and a WCST-Cat score of > or =3.5 was assumed to be employable. Sensitivity analysis was performed for key values. RESULTS: The base case per-patient cost of risperidone and haloperidol was 6,422 US dollars and 4,989 US dollars, respectively, and the number of employable persons was 3,258 (32.58%) and 2,517 (25.17%), respectively, which translates to an incremental CE ratio of 19,609 US dollars per employable person for risperidone. Risperidone remained cost increasing and had higher number of employable persons over all the ranges used in the sensitivity analysis. The incremental CE ratio ranged from a high of 1,000,000 US dollars to a low of 3,000 US dollars per employable persons when the rates of clinical stability for risperidone and haloperidol therapy were varied. CONCLUSION: Gains from earning rates for employed persons with schizophrenia, savings in informal caregiver costs and other human benefits could justify an incremental cost of 19,609 US dollars for each additional employable person prescribed risperidone. PMID- 12892866 TI - Cognitive behavioural group treatment for social anxiety in schizophrenia. AB - Anxiety symptoms reported by individuals with schizophrenia have been traditionally seen as symptoms associated with the principal disorder and therefore not requiring special attention. The primary aim of this paper is to therapeutically target social anxiety symptoms in individuals with schizophrenia in order to determine the effectiveness of the cognitive behavioural group treatment model as an intervention for social anxiety in this participant group. Thirty-three individuals with schizophrenia and co-morbid social anxiety were allocated to a group-based cognitive behaviour (CBGT) intervention or waitlist control (WLC). Baseline, completion and follow-up ratings consist of measures of social anxiety: the Brief Social Phobia Scale (BSPS), Brief Fear of Negative Evaluation scale (BFNE) and the Social Interaction Anxiety Scale (SIAS); measures of general psychopathology: the Calgary Depression Scale for Schizophrenia (CDSS) and Global Severity Index (GSI) from the Brief Symptom Inventory (BSI); and the Quality of Life, Enjoyment and Satisfaction Questionnaire (QLESQ). Pre- and post treatment measures were subjected to statistical evaluation. All outcome measures displayed statistical improvement in the intervention group compared with no change in the control group. These treatment gains were maintained at follow-up. CBGT for social anxiety in schizophrenia was demonstrated to be effective as an adjunctive treatment for this population. PMID- 12892867 TI - No relationship between risk of schizophrenia and prenatal exposure to stress during the Six-Day War or Yom Kippur War in Israel. AB - AIM: Maternal stress during pregnancy is a possible risk factor for schizophrenia in the offspring. Using data from the Israel Psychiatric Registry we examined the impact of wars in Israel. METHOD: Retrospective birth cohort study. RESULTS: Relative risks for cohorts exposed to Six-Day War and Yom Kippur War were 0.98 (95% CI: 0.85-1.13) and 1.00 (0.86-1.16). CONCLUSION: The evidence for maternal stress as a risk factor for schizophrenia remains insufficient. PMID- 12892868 TI - Development and validation of a patient-based health-related quality of life questionnaire in schizophrenia: the S-QoL. AB - We developed a self-administered instrument to assess health-related quality of life (HRQL) among people with schizophrenia. The S-QoL, based on Calman's approach to the subject's point of view, is a multidimensional instrument that is sensitive to change. The scale is a 41-item questionnaire with eight subscales (psychological well-being, self-esteem, family relationships, relationships with friends, resilience, physical well-being, autonomy and sentimental life) and a total score. In-depth interviews with patients determined the pertinent issues for item development. The validation study, performed with 207 patients, showed high internal consistency reliability, reproducibility and responsiveness. Construct validity was confirmed using established clinical and HRQL measures. S QoL covers domains that differ from areas tapped in other measures, with greater responsiveness. The S-QoL is an efficient instrument for the measurement of the impact of schizophrenia on individuals' lives. PMID- 12892869 TI - Neuroleptic withdrawal in treatment-resistant patients with schizophrenia: tardive dyskinesia is not associated with supersensitive psychosis. AB - The objective of this retrospective study was to determine whether tardive dyskinesia (TD) represents a risk factor for supersensitive psychosis (SS) by assessing the effect of medication withdrawal on ratings of psychopathology for 30 days following discontinuation of antipsychotic medication in patients with and without TD. The subjects were 101 treatment-resistant patients with schizophrenia who had been admitted to the inpatient service of Neuroscience Research Hospital (NRH), National Institute of Mental Health, between 1982 and 1994 to undergo studies involving discontinuation of antipsychotic medication. Patients were rated independently on a daily basis on the 22-item Psychiatric Symptom Assessment Scale (PSAS), an extended version of the Brief Psychiatric Rating Scale (BPRS). The overall frequency of TD was 35.6%. Tardive dyskinesia patients were older (p < 0.0006) and had suffered from schizophrenia for a longer time (p < 0.003) than No-TD patients. Repeated measure ANOVA revealed a "time" effect for all subgroups studied. The interaction TD x time, however, was not statistically significant for any of the clusters. Within-group analysis revealed significant differences against baseline for measures of positive symptoms, negative symptoms and abnormal involuntary movements in the No-TD group 3 and 4 weeks after antipsychotic withdrawal. In the TD group, however, the changes were observed only at 4 weeks following antipsychotic discontinuation in just two of the positive symptoms cluster. Between-group analyses revealed that, at baseline, the Mannerisms cluster (abnormal involuntary movements) was significantly higher in the TD group (p < 0.05). No significant differences were observed between any of the remaining clusters at baseline or at different times following drug withdrawal. In conclusion, the relationship between SS and TD could not be confirmed in a cohort of patients with treatment-resistant schizophrenia. In the present study, patients with no TD seemed to deteriorate faster than patients with TD in terms of psychopathology and abnormal involuntary movements. It is possible that both group of patients may undergo supersensitive receptor changes, and that these changes may be more pronounced but potentially reversible in the group without TD. PMID- 12892871 TI - The evolution of symptoms in the early course of non-affective psychosis. AB - Most previous studies investigating the factor structure of psychosis have focussed on chronic samples. First episode samples with longitudinal follow up are few. To investigate the stability and validity of symptom factors, a sample of 257 patients with DSM IV nonaffective psychoses were assessed using the PANSS during the acute first episode and at 3- and 18-month follow up. Exploratory factor analysis of the changes in PANSS item scores over time gave a five-factor solution. This was consistent with the solutions to factor analyses at the initial assessment and each of the follow-ups. However, there was progression over follow-up. Confirmatory factor analysis demonstrated that symptom ratings at 18-month follow-up fitted the models from existing research, in relatively chronic samples, better than the ratings at the initial assessment. A psychomotor poverty factor showed most stability over time and a positive symptom factor most change. Factors showed different associations with demographic and external variables, further supporting their validity. PMID- 12892872 TI - Neurological soft-signs and minor physical anomalies in schizophrenia: differential transmission within families. AB - Markers of vulnerability have been identified in schizophrenia, and among them, neurological soft-signs (NSS) and minor physical anomalies (MPAs) also seem to occur in biological relatives. The similarities of these developmental markers within families may depend on either genetic or non-genetic factors. The aim of the study was to investigate the intra-familial similarities of NSS and MPAs within 18 nuclear families (18 probands with schizophrenia and 36 of their non psychotic parents). A general linear model showed similarities within families for NSS (intra-class coefficient [ICC] = 0.64; F = 2.6; df = 17.17; p = 0.02) but not for MPAs (ICC = -0.10; F = 0.7; df = 17.17; ns). We thus found a direct evidence for the intra-familial transmission of NSS but not of MPAs, suggesting that this morphological phenotypic trait could be more dependent on epigenetic influences. PMID- 12892870 TI - The InterSePT scale for suicidal thinking reliability and validity. AB - BACKGROUND: The InterSePT Scale for Suicidal Thinking (ISST) is a 12-item instrument for the assessment of current suicidal ideation in patients with schizophrenia and schizoaffective disorders. We report the psychometric characteristics of this new scale based on two studies. METHOD: In Study 1, 22 inpatients with schizophrenia and schizoaffective disorders, who had recently attempted suicide or engaged in suicidal ideation, were rated by three trained independent raters to examine interrater reliability. In Study 2, a total of 980 patients with schizophrenia or schizoaffective disorder with a history of suicidal ideation in the past 36 months were enrolled in a 2-year industry sponsored suicide prevention study. At baseline, these patients were administered the ISST and the Clinical Global Impression Scale for Severity of Suicidality (CGI-SS) by the Principal Investigator (PI) and by a blinded rater (BR), who also administered the Positive and Negative Symptom Scale (PANSS), the Calgary Depression Scale (CDS), and the Scale of Functioning (SOF). Indices of internal reliability, construct and discriminant validity were examined. RESULTS: The intraclass correlation coefficient (ICC) for the total ISST score for the 22 subjects in Study 1 was 0.90 and mean weighted item kappa coefficients ranged from 0.66 to 0.92. In Study 2, internal reliability (Cronbach alpha) was high, ranging from 0.86 to 0.89 for the individual items, and the overall Cronbach alpha coefficient for all items was 0.88. The ISST (PI) total score was highly correlated with the CGI-SS by the blind rater (r = 0.61, p < 0.0001). ISST total scores significantly differentiated the different levels of CGI-SS (F = 519.2; p < 0.0001). Results of construct and discriminant validity analyses are also presented. CONCLUSION: The ISST is a reliable and valid instrument for the assessment of current suicidal thinking in patients with schizophrenia and schizoaffective disorder by both clinicians and researchers. PMID- 12892873 TI - Acute and maintenance ECT with flupenthixol in refractory schizophrenia: sustained improvements in psychopathology, quality of life, and social outcomes. AB - OBJECTIVE: To determine the effects of ECT combined with antipsychotic medication therapy on psychopathology, quality of life, and social functioning in patients with refractory schizophrenia. METHOD: An open acute (Phase I) and maintenance (Phase II) study of the combination of ECT and flupenthixol in the treatment of 46 schizophrenic patients who were nonresponsive to antipsychotic medication from at least two different classes. Scales used: the Brief Psychiatric Rating Scale (BPRS), the Quality of Life Scale (QLS), Social and Occupational Functioning Assessment Scale (SOFAS), Global Assessment of Functioning (GAF), and Mini-Mental State Exam (MMSE). The duration of Phase II was 1 year. RESULTS: In Phase I, there were marked reductions in the BPRS scores, and substantial increases in the QLS, SOFAS, GAF, and MMSE scores. During Phase II, the BPRS negative symptoms worsened but remained improved from baseline. Changes in other outcome measures were negligible. CONCLUSION: ECT and MECT combined with flupenthixol were effective in improving psychopathology in patients refractory to antipsychotic medication alone. Ratings of psychopathology, quality of life, and social functioning all improved in Phase I and were generally sustained during Phase II in patients who had remitted. PMID- 12892874 TI - Evaluating sexual function in patients with treatment-resistant schizophrenia. PMID- 12892875 TI - Dopamine receptor hetero-oligomerization in the pathogenesis of schizophrenia: qualitative as opposed to quantitative notions of dopaminergic receptors. PMID- 12892876 TI - An oxidized form of apomorphine fails to induce stereotypy. PMID- 12892877 TI - Bacteria that express lateral flagella enable dissection of the multifunctional roles of flagella in pathogenesis. AB - Flagella are much more than organelles of locomotion and have multiple roles that contribute to pathogenesis. Bacteria, such as Vibrio parahaemolyticus and Aeromonas spp., that possess two distinct flagellar systems (a polar flagellum for swimming in liquid and lateral flagella for swarming over surfaces) are relatively uncommon and provide ideal models for the independent investigation of the contributions of these different types of motility and other flagellar functions to virulence and how they are controlled. Studies with the above organisms have already increased our understanding of how bacteria sense and colonize surfaces forming biofilms that enable them to survive and persist in hostile environments. These insights are helping to identify possible new targets for novel antimicrobials that will both prevent or disrupt these processes and enhance the effectiveness of existing antibiotics. Aeromonas lateral flagella, in addition to mediating swarming motility, appear to be adhesins in their own right, contribute to microcolony formation and efficient biofilm formation on surfaces, and possibly facilitate host cell invasion. It is, therefore, likely that the ability to express lateral flagella is a significant virulence determinant for the Aeromonas strains able to cause persistent and dysenteric infections in the gastrointestinal tract, but further work is needed to establish this. PMID- 12892878 TI - Subcellular localization of 14-3-3 proteins in Toxoplasma gondii tachyzoites and evidence for a lipid raft-associated form. AB - A polyclonal antibody was raised against a Toxoplasma gondii 14-3-3-gluthatione S transferase fusion protein obtained by cloning a 14-3-3 cDNA sequence determined from the T. gondii database. This antibody specifically recognized T. gondii 14-3 3 without any cross-reaction with mammalian proteins. Immunofluorescence microscopy studies of the tachyzoites or the T. gondii-infected cells suggested cytosolic and membranous localizations of 14-3-3 protein. Different subcellular fractions were prepared for electrophoresis analysis and immunodetection. 14-3-3 proteins were found in the cytosol, the membrane fraction and Triton X-100 resistant membranes. Two 14-3-3 isoforms were detected. The major one was mainly cytoplasmic and to a lesser extent membrane-associated, whereas the minor isoform was associated with the detergent-resistant lipid rafts. PMID- 12892879 TI - A mutant Bacillus subtilis gamma-glutamyltranspeptidase specialized in hydrolysis activity. AB - gamma-Glutamyltranspeptidase (GGT) catalyzes the hydrolysis of gamma-glutamyl compounds and the transfer of their gamma-glutamyl moieties to amino acids and peptides. The transpeptidation activity of Bacillus subtilis GGT is about 10-fold higher than its hydrolysis activity. In B. subtilis GGT, substitution of Asp-445 with Ala abolished its transpeptidation activity. The specific activity for hydrolysis of D445A GGT was 40.2% of that of the wild-type GGT. The K(m) value for L-glutamine was 15.3 mM. D445A GGT was salt tolerant like the wild-type GGT. These results indicate that D445A GGT will be highly useful as a 'glutaminase' in food industry. PMID- 12892880 TI - Helicobacter pylori strain ATCC700392 encodes a methyl-accepting chemotaxis receptor protein (MCP) for arginine and sodium bicarbonate. AB - Helicobacter pylori ATCC43504 responds chemotactically to aspartic acid and serine, but not to arginine, nor to sodium bicarbonate. In contrast, H. pylori ATCC700392 (strain 26695) shows chemotaxis to all four attractants. Open reading frame HP0099 from H. pylori 26695 is predicted to encode one of three methyl accepting chemotaxis receptor proteins (MCPs). When Escherichia coli is transformed with a plasmid carrying HP0099 from strain 26695, the recombinants acquire chemotaxis to arginine, bicarbonate, and urea. In H. pylori 43504, the HP0099 gene is interrupted with a mini-IS605 insertion, which accounts for its inability to recognize arginine and bicarbonate as attractants. Together, these results argue that the H. pylori HP0099 gene encodes an MCP for arginine and bicarbonate. PMID- 12892882 TI - New integration vector using a cellulase gene as a screening marker for Lactobacillus. AB - The new integration vector for Lactobacillus, pJC4, was developed using the extracellular endoglucanase A gene (celA) of Clostridium thermocellum as a screening marker. pJC4 was transformed into four Lactobacillus species, Lb. johnsonii, Lb. gasseri, Lb. bulgaricus, and Lb. plantarum. In each species, the pJC4 integrants were easily and accurately detected by the appearance of a clear halo on a cellulase screening plate without any false transformants. Polymerase chain reaction and Southern hybridization indicated that all transformants with clear halos contained pJC4 in their chromosomal DNAs. The celA gene could be a useful screening marker for other lactic acid bacteria. PMID- 12892881 TI - Kakadumycins, novel antibiotics from Streptomyces sp NRRL 30566, an endophyte of Grevillea pteridifolia. AB - An endophytic streptomycete (NRRL 30566) is described and partially characterized from a fern-leaved grevillea (Grevillea pteridifolia) tree growing in the Northern Territory of Australia. This endophytic streptomycete produces, in culture, novel antibiotics - the kakadumycins. Methods are outlined for the production and chemical characterization of kakadumycin A and related compounds. This antibiotic is structurally related to a quinoxaline antibiotic, echinomycin. Each contains, by virtue of their amino acid compositions, alanine, serine and an unknown amino acid. Other biological, spectral and chromatographic differences between these two compounds occur and are given. Kakadumycin A has wide spectrum antibiotic activity, especially against Gram-positive bacteria, and it generally displays better bioactivity than echinomycin. For instance, against Bacillus anthracis strains, kakadumycin A has minimum inhibitory concentrations of 0.2-0.3 microg x ml(-1) in contrast to echinomycin at 1.0-1.2 microg x ml(-1). Both echinomycin and kakadumycin A have impressive activity against the malarial parasite Plasmodium falciparum with LD(50)s in the range of 7-10 ng x ml(-1). In macromolecular synthesis assays both kakadumycin A and echinomycin have similar effects on the inhibition of RNA synthesis. It appears that the endophytic Streptomyces sp. offer some promise for the discovery of novel antibiotics with pharmacological potential. PMID- 12892883 TI - The thiocarbamate-inducible Rhodococcus enzyme ThcF as a member of the family of alpha/beta hydrolases with haloperoxidative side activity. AB - Purified thiocarbamate-inducible ThcF of Rhodococcus erythropolis NI86/21, overexpressed in Escherichia coli, displayed several characteristics of the HASH family of enzymes that groups prokaryotic proteins of the alpha/beta hydrolase superfamily possessing serine-dependent hydrolase and/or haloperoxidase activity. Kinetic analysis of bromination and ester hydrolysis revealed a low affinity of ThcF for model substrates. Sulfoxidation of thiocarbamates was demonstrated but probably represents a side activity due to peroxoacid generation by the enzyme. The thcF-linked thcG gene, encoding a LAL-type regulator, triggers expression of thcF in Rhodococcus. The tandem gene organization thcG-thcF is conserved in the thiocarbamate-degrading strain Rhodococcus sp. B30. It is proposed that HASH enzymes may be involved in the metabolism of plant-derived compounds. PMID- 12892884 TI - Description of five mutants of the cyanobacterium Anabaena sp strain PCC 7120 affected in heterocyst differentiation and identification of the transposon tagged genes. AB - When growing on N(2) as sole nitrogen source, the filamentous cyanobacterium Anabaena sp. PCC 7120 forms N(2) fixing heterocysts in a semi-regular pattern. To identify genes involved in heterocyst differentiation we characterised five transposon-generated mutants that were not able to form mature heterocysts. After recovering the transposon together with the flanking region of the Anabaena chromosome the affected genes were identified. Four of the genes could be involved in formation of the heterocyst-specific envelope: alr2887, encoding a probable outer membrane efflux protein, alr3698, a glycosyl transferase, all4388, a putative periplasmic polysaccharide export protein and alr5357, the formerly described gene hglB/hetM, encoding a fatty-acid synthetase. Another gene, all0049/mutS2, may be important in one of the genome rearrangements that occur during heterocyst differentiation. By transcriptional fusion to reporter genes luxAB differential expression of alr2887, alr3698 and alr5357 could be monitored during heterocyst differentiation. PMID- 12892885 TI - Characterisation of a new species of Pythium isolated from a wheat field in northern France and its antagonism towards Botrytis cinerea causing the grey mould disease of the grapevine. AB - A new species, Pythium bifurcatum, isolated from soil samples taken from a wheat field in Lille in northern France is described here. The oomycete occurred thrice out of 50 samples. The type specimen is F-91, which is a slow-growing saprophyte living on vegetable debris and which can be recognised by its antheridial as well as oogonial characteristics, which are different from other known species of Pythium. When grown together with Botrytis cinerea, the causal agent of the grey mould disease of the grapevine, Pythium bifurcatum shows a pronounced antagonism and suppresses its growth. Morphological features of this new species, its antagonism to B. cinerea, the sequences of the ITS region of its nuclear ribosomal DNA, and its comparison with related species are discussed in this article. PMID- 12892886 TI - Detection of bacteriophage VSH-1 svp38 gene in Brachyspira spirochetes. AB - VSH-1 is a mitomycin C-inducible, non-lytic, phage-like agent that packages random 7.5-kb fragments of the Brachyspira hyodysenteriae genome. VSH-1 is the first recognized mechanism for gene transfer between B. hyodysenteriae cells. To analyze the distribution of VSH-1 among spirochetes, a 344-bp probe for gene svp38, encoding the VSH-1 major head protein, was amplified by polymerase chain reaction and used in Southern blot hybridizations with genomic DNA from various spirochete genera. The svp38 probe hybridized to a 40-kb SalI-SmaI fragment of the B. hyodysenteriae B78(T) chromosome, indicating VSH-1 DNA insertion into the chromosome at a unique site. Restriction endonuclease digested DNAs of 27 spirochete strains representing six Brachyspira species (B. hyodysenteriae, B. innocens, B. pilosicoli, B. murdochii, B. intermedia, B. alvinipulli) contained a single fragment hybridizing with the svp38 probe. DNAs from spirochete species of the genera Treponema, Spirochaeta, Borrelia, and Leptospira did not hybridize with the probe. VSH-1-like agents appear to be widely distributed among Brachyspira species and, as has been demonstrated for B. hyodysenteriae, may serve as useful gene transfer agents for those other species. PMID- 12892887 TI - Molecular cloning of a phospholipase D gene from Aspergillus nidulans and characterization of its deletion mutants. AB - We cloned a gene pldA encoding a protein containing phospholipase D (PLD) motifs from a filamentous fungus Aspergillus nidulans. The deduced protein product of pldA consists of 833 amino acids and contains four conserved regions of a PLD gene family. Deletion mutants of pldA grew and formed conidia in a normal manner. Although PLD and transphosphatidylation activities against phosphatidylcholine of the mutant cell extract did not change, the Ca(2+)-dependent PLD activity against phosphatidylethanolamine was significantly reduced, but not in the wild-type cell extract. This activity was markedly enhanced by high osmotic growth conditions in the wild-type cells, and pldA of A. nidulans likely encodes a Ca(2+)-dependent phosphatidylethanolamine-hydrolyzing PLD. PMID- 12892888 TI - Identification of the CysB-regulated gene, hslJ, related to the Escherichia coli novobiocin resistance phenotype. AB - The cysB gene product is a LysR-type regulatory protein required for expression of the cys regulon. cysB mutants of Escherichia coli and Salmonella, along with being auxotrophs for the cysteine, exhibit increased resistance to the antibiotics novobiocin (Nov) and mecillinam. In this work, by using lambdaplacMu9 insertions creating random lacZ fusions, we identify a gene, hslJ, whose expression appeared to be increased in cysB mutants and needed for Nov resistance. Measurements of the HSLJ::lacZ gene fusion expression demonstrated that the hslJ gene is negatively regulated by CysB. In addition we observe the negative autogenous control of HslJ. When the control imposed by CysB is lifted in the cysB mutant, the elevation of Nov resistance can be achieved only in the presence of wild-type hslJ allele. A double cysB hslJ mutant restores the sensitivity to Nov. Overexpression of the wild-type HslJ protein either in a cysB(+) or a cysB(-) background increases the level of Nov resistance indicating that hslJ product is indeed involved in accomplishing this phenotype. The HSLJ::OmegaKan allele encodes the C-terminaly truncated mutant protein HslJ Q121Ter which is not functional in achieving the Nov resistance but when overexpressed induces the psp operon. Finally, we found that inactivation of hslJ does not affect the increased resistance to mecillinam in cysB mutants. PMID- 12892889 TI - Diverse effects of ascorbic acid and palmitoyl ascorbate on Helicobacter pylori survival and growth. AB - Among many antioxidants used in the food, pharmaceutical and cosmetic industries, ascorbic acid (AA) is one of the most important. AA has been suggested to decrease the risk of gastric disease (gastritis, duodenal ulcer, and carcinoma) by direct action on Helicobacter pylori. However, there are limited studies on the possible role of AA and its derivatives such as palmitoyl ascorbate (PA) on the growth and survival of H. pylori. In the present study it was demonstrated in vitro that AA in the concentration range 10-20 mg x ml(-1) (50-100 mM) inhibited H. pylori growth in liquid medium under microaerophilic conditions. In contrast, under aerobic conditions AA in the concentration range 2-20 mg x ml(-1) (10-100 mM) significantly increased the survival of H. pylori presumably eliminating the toxic effect of reactive oxygen species on bacterial cells. The hydrophobic derivative of AA, PA (a food antioxidant), demonstrated a strong antibacterial effect, under both aerobic and microaerophilic conditions in the concentration range 0.04-0.4 mg x ml(-1) (0.1-1.0 mM). This effect was also tested on other bacterial strains: Escherichia coli, Proteus vulgaris, Proteus mirabilis, Pseudomonas aeruginosa, Enterococcus faecalis, Bacillus cereus, Bacillus subtilis, Staphylococcus aureus, Staphylococcus epidermidis, Clostridium sporogenes and Campylobacter jejuni. Among these bacterial strains, PA showed a similar inhibitory effect on B. cereus and B. subtilis as observed with H. pylori. The results suggest that PA may be considered an important AA derivative in eradication of H. pylori in vitro and in vivo and to decrease the risk for gastric diseases. PMID- 12892890 TI - Nitrogenase activity of Herbaspirillum seropedicae grown under low iron levels requires the products of nifXorf1 genes. AB - Herbaspirillum seropedicae strains mutated in the nifX or orf1 genes showed 90% or 50% reduction in nitrogenase activity under low levels of iron or molybdenum respectively. Mutations in nifX or orf1 genes did not affect nif gene expression since a nifH::lacZ fusion was fully active in both mutants. nifX and the contiguous gene orf1 are essential for maximum nitrogen fixation under iron limitation and are probably involved in synthesis of nitrogenase iron or iron molybdenum clusters. PMID- 12892891 TI - Molecular characterization of the Candida albicans LYS5 gene and site-directed mutational analysis of the PPTase (Lys5p) domains for lysine biosynthesis. AB - The LYS2 and LYS5 genes of the pathogenic yeast Candida albicans are required for the alpha-aminoadipate reductase (AAR) reaction in the lysine biosynthetic pathway. The LYS2 encodes an apo-AAR (Lys2p) and the LYS5 encodes a phosphopantetheinyl transferase (PPTase) for the post-translational activation of AAR. Our cloned C. albicans LYS5 gene encodes a 38.4 kDa PPTase which is 27% identical and 43% similar to the Saccharomyces cerevisiae Lys5p. Sequence alignment of Lys5p with other PPTases reveals highly conserved putative PPTase domains including the Core 3, WXXKESXXK (residues 194-202). Recombinant Lys5p expressed in Escherichia coli activates C. albicans Lys2p for the AAR activity and also activates AARs from S. cerevisiae and to a lesser extent Schizosaccharomyces pombe. Site-directed mutational analyses reveal glutamic acid 198 in the Lys5p Core 3 as essential for the activation of recombinant Lys2p AAR activity. Other conserved amino acids were also analyzed for their influence on Lys5p PPTase activity. Our results demonstrate cloning of the LYS5 gene, expression of Lys5p, in vitro Lys2p activation model and characterization of the functional motifs of the C. albicans PPTase. PMID- 12892892 TI - A study on microbial diversity in different cultivars of Brassica napus in relation to its wilt pathogen, Verticillium longisporum. AB - Oilseed rape (Brassica napus) is one of the major oilseed crops in the world but is vulnerable to attack by many pathogens and insect pests. In addition to the host plant genotype, micro-organisms present in the rhizosphere and within plant tissues affect the susceptibility to plant pathogens. While rapid progress has been achieved concerning the concept of plant resistance genes, information on the role of the microbial community in plant protection is less apparent. We have studied the endophytic bacterial populations present in different tissues of oilseed rape and also analysed several cultivars (Express, Libraska, Maluka and Westar), which differ in their susceptibility to the wilt pathogen Verticillium longisporum. The population diversity was studied using agar plating assay, fatty acid methyl ester analysis and functional characterisation of isolated strains. Our work shows that already in the seeds there exists diversity in populations as well as in the total microbial load between two of the four tested cultivars. About 50% of the strains isolated from cultivars Express and Libraska showed moderate to strong direct inhibition of V. longisporum. The diversity of the endophytic flora isolated from oilseed rape and its implications in crop protection are discussed. PMID- 12892893 TI - Conversion of Shigella flexneri serotype 2a to serotype Y in a shigellosis patient due to a single amino acid substitution in the protein product of the bacterial glucosyltransferase gtrII gene. AB - Conversion of serotype from 2a to Y was demonstrated with five Shigella flexneri isolates recovered from an infected patient. When introduced into the serotype Y isolate, the glucosyltransferase (gtr) II gene of the serotype 2a isolate is capable of inducing the conversion from serotype Y to 2a. In contrast, the gtrII of the serotype Y isolate lacks the capacity to change serotype, resulting from a Cys-->Tyr substitution in its predicted protein sequence. The protein product of the gtrII gene was detected. This is the first report of serotype conversion of S. flexneri in humans, and successful detection of the protein product from a gtr gene. PMID- 12892894 TI - Purification and characterization of a novel lignin peroxidase from white-rot fungus Phanerochaete sordida YK-624. AB - We characterized kinetics and substrate oxidation of a novel lignin peroxidase (YK-LiP) isolated from white-rot fungus Phanerochaete sordida YK-624. YK-LiP enzyme was identified and purified to homogeneity by anion-exchange chromatography and gel permeation chromatography. The molecular mass of YK-LiP was approximately 50 kDa, and the absorption spectrum of YK-LiP was almost the same as that of the LiP (Pc-LiP) from Phanerochaete chrysosporium. Steady-state kinetics of veratryl alcohol oxidation by YK-LiP (unlike that by Pc-LiP) revealed a bi-reactant sequential mechanism, although reactivity of YK-LiP to various monomeric substituted aromatic compounds was similar to that of Pc-LiP. Degradation of dimeric lignin model compounds was more effective by YK-LiP than by Pc-LiP, and the oxidation rate of sinapyl alcohol oligomer by YK-LiP was much faster than that by Pc-LiP. PMID- 12892895 TI - msDNA-St85, a multicopy single-stranded DNA isolated from Salmonella enterica serovar Typhimurium LT2 with the genomic analysis of its retron. AB - Bacterial reverse transcriptase is responsible for the production of a small satellite DNA-RNA complex called multicopy single-stranded DNA (msDNA) that has been found in a wide variety of Gram-negative bacteria. Here we describe the isolation and characterization of a novel msDNA, msDNA-St85, from Salmonella enterica serovar Typhimurium LT2. We determined the nucleotide sequence of msDNA St85 and the location of retron-St85 on the chromosome that is responsible for msDNA-St85 production by analyzing the complete genomic sequence of S. typhimurium LT2. It was found that the G+C content and the codon usage of retron St85 were significantly different from those of the S. typhimurium genome, indicating that retron-St85 was probably acquired recently in this bacterium. This is the first report for identification of an msDNA in the genus Salmonella with the complete description and analysis of its retron. PMID- 12892896 TI - Chitin synthase III activity, but not the chitin ring, is required for remedial septa formation in budding yeast. AB - Chitin is a minor but essential component of the Saccharomyces cerevisiae cell wall. In wild-type, chitin synthase II is required for the formation of primary septa and chitin synthase III (CSIII) is not essential. However, in chs2 mutants CSIII becomes essential for the formation of aberrant septa. We examined which of two CSIII functions, the formation of a chitin ring at bud emergence or of chitin in the remedial septa, was required for viability. By using cell cycle synchronization in combination with nikkomycin Z, a specific inhibitor of CSIII, we inhibited chitin synthesis in a chs2 mutant, during formation of either the ring or the remedial septa. The results show that only synthesis of the chitin during aberrant septa formation is essential for viability. Thus, the unique function of the chitin ring seems to be maintenance of the integrity of the mother-bud neck, as we recently found, and the importance of chitin in septum closure, both in normal and abnormal situations, is underlined. PMID- 12892897 TI - Controlled intra- or extracellular production of staphylococcal nuclease and ovine omega interferon in Lactococcus lactis. AB - A system for controlled targeting of heterologous protein was developed in the food-grade bacterium Lactococcus lactis. It is composed of the L. lactis strain NZ9000 and of two broad host range expression vectors pCYT:Nuc and pSEC:Nuc for, respectively, cytoplasmic and secreted staphylococcal nuclease (Nuc) nisin inducible production. The level of intracellular production of Nuc measured with pCYT:Nuc (3 mg x l(-1)) is significantly lower than the one obtained with pSEC:Nuc ( approximately 20 mg x l(-1)). The secretion efficiency (SE) of Nuc is estimated to be approximately 70%, corresponding to approximately 15 mg of secreted Nuc x l(-1). Furthermore, we established that Nuc production continued in L. lactis 10 h after a 1-h nisin-pulse induction. This system was then used for intra- and extracellular production of a protein of therapeutical interest in L. lactis, the ovine interferon-omega (IFN-omega). The SE and the quantity of secreted active IFN-omega were evaluated respectively to be approximately 70% and approximately 1 mg x l(-1) ( approximately two-fold higher than the cytoplasmic form). PMID- 12892898 TI - G-CSF immunotherapy for treatment of acute disseminated murine melioidosis. AB - Burkholderia pseudomallei, the causative agent of melioidosis, is an important pathogen in tropical regions of Australia and Southeast Asia. Antibiotic therapy can be ineffective in patients with acute septicaemic melioidosis. It has been proposed that adjunctive immunotherapy using granulocyte colony stimulating factor (G-CSF) combined with antibiotics may provide an alternative approach to antibiotics alone. We have developed a murine model for melioidosis that allows novel treatment approaches to be investigated. This study looked at the potential for murine G-CSF therapy both alone and as an adjunct in the treatment of acute disseminated B. pseudomallei infection in BALB/c mice. A number of therapeutic variables involving ceftazidime and recombinant murine G-CSF were studied. Surviving mice were sacrificed and splenic bacterial loads were determined. Combining recombinant murine G-CSF with ceftazidime offered no advantage over ceftazidime alone. Pre-treatment with recombinant murine G-CSF did not demonstrate a significant benefit. This would suggest that adjunct immunotherapy using G-CSF is of limited benefit. PMID- 12892899 TI - Differential expression of the natural antimicrobials, beta-defensins 3 and 4, in human endometrium. AB - beta-Defensins are small cationic molecules that have antimicrobial actions against bacteria, fungi and viruses and contribute to mucosal immune responses at epithelial sites. The female reproductive tract is an important site of defensin production and innate defences are crucial to the preservation of fertility and successful pregnancy. This study details the expression of the recently characterized defensins, HBD3 and 4, in human endometrium. Using real-time quantitative RT-PCR, we have shown that HBD3 mRNA expression is highest during the secretory phase of the menstrual cycle while HBD4 mRNA levels peak in the proliferative phase. Both antimicrobials are expressed by endometrial epithelium. Exogenous steroid hormones in the form of the combined oral contraceptive pill (COCP) alter expression of both defensins in vivo, while treatment of endometrial explants with progesterone in vitro does not alter expression of HBD3 or HBD4. In in vitro cultures of primary endometrial epithelial cells, HBD3 mRNA expression is upregulated by treatment with inflammatory molecules including IL-1 beta+TNF alpha, IFN gamma and phorbol ester. HBD4 mRNA was not expressed in these primary cell cultures. These results show that the human endometrium expresses both HBD3 and HBD4 in a cycle-dependent manner. These natural antimicrobials will contribute to innate defences present in human endometrium protecting against uterine infection. Expression is altered as a result of hormonal contraceptive use and this may contribute to differential infection rates in COCP users relative to non-users. In addition, expression of HBD3 will be upregulated during infection allowing an increased innate immune response at this time. PMID- 12892900 TI - 17 beta-estradiol induces L-type Ca2+ channel activation and regulates redox function in macrophages. AB - 17 beta-estradiol induces rapid effects on cells of the immune system via plasma membrane surface receptor but the ways in which delayed signals involving intracellular receptors affect the same functions are not well understood. To study the delayed but sustained events in estradiol signaling, we have investigated macrophage Ca(2+) signaling, detected specific Ca(2+) ion channel activated and found a relationship between intracellular calcium [Ca(2+)](i) and macrophage release of reactive oxygen species (ROS) and nitric oxide (NO) during delayed 17 beta-estradiol activity. We found evidence of additional effect of estradiol on capacitative entry of Ca(2+), Ca(2+) entry through L-type channel and a direct relationship between [Ca(2+)](i) and generation of ROS and NO. This study demonstrates that 17 beta-estradiol exhibits a delayed phase of Ca(2+) influx involving L-type channel and regulates macrophage immune redox function. PMID- 12892901 TI - Elevated C-reactive protein levels during first trimester of pregnancy are indicative of preeclampsia and intrauterine growth restriction. AB - C-reactive protein (CRP) is a marker of tissue damage and inflammation. Maternal levels of CRP are elevated in overt preeclampsia, but there is still debate about its use as a predictive marker for preeclampsia during the first and second trimesters of pregnancy. In this study, we measured CRP levels during the first trimester of pregnancy in women who later developed preeclampsia or gave birth to a growth-restricted baby. In total, 107 women from a low-risk population participated in the study, six women developed preeclampsia and nine gave birth to a growth-restricted baby. Although there is a large overlap in measured CRP levels between the three groups, mean CRP levels were significantly elevated in women who later developed preeclampsia (P=0.031) or delivered a growth-restricted baby (P=0.041) when compared with women from the control group, matched for maternal and gestational age, parity, and gravidity. This study shows that in a low-risk population, CRP levels are already elevated between weeks 10 and 14 in pregnant women who develop preeclampsia or deliver a growth-restricted baby. PMID- 12892902 TI - Total white blood cell counts and LPS-induced TNF alpha production by monocytes of pregnant, pseudopregnant and cyclic rats. AB - Pregnancy in the rat may be associated with an activated innate immune system. Therefore, we investigated monocyte function as well as total white blood cell (WBC) counts during the follicular phase of the ovarian cycle, pregnancy and pseudopregnancy in the rat. Rats were equipped with a permanent jugular vein cannula, and 0.43 ml blood samples were taken from this cannula during the 4 days of the regular oestrus cycle of the rat (n=12). Thereafter, six rats were rendered pregnant, and the other six rats were rendered pseudopregnant according to standard methods. Blood samples were withdrawn from the cannula on days 4, 7 and 11 of pseudopregnancy and on days 4, 7, 11 and 20 of pregnancy. From each blood sample, 0.4 ml was stimulated with lipopolysaccharide (LPS) and monocyte intracellular cytokine production measured using flow cytometry. 30 microl of the blood was used to measure WBC counts and differential WBC counts. The results showed that the number of WBC was significantly increased only on day 11 of pregnancy compared with the follicular phase, and that this was due to the increased numbers of polymorphonuclear (PMN) cells. The percentage of TNF alpha producing monocytes was increased on all days of pseudopregnancy and on day 11 of pregnancy. The fact that the percentage of monocytes producing TNF alpha upon an LPS stimulus was increased during the post-implantation phase of pregnancy and during pseudopregnancy as compared to the follicular phase may indicate that these conditions are proinflammatory conditions. For the post-implantation phase of pregnancy, this is once more stressed by the increased numbers of WBC and PMN. PMID- 12892903 TI - Production of the antiviral proteins 2'5'oligoadenylate synthetase, PKR and Mx in interstitial cells and spermatogonia. AB - We report an in vitro analysis of the spatial pattern of production of three antiviral proteins (2'5'oligoadenylate synthetase, 2'5'AS; double-stranded RNA activated protein kinase, PKR; and Mx protein, Mx) in the rat testis, in basal conditions and following stimulation with interferon (IFN) or Sendai virus. The two major constituents of interstitial tissue--Leydig cells and macrophages- constitutively produce 2'5' oligoadenylate synthetase (2'5'AS), PKR and Mx. Production of an isoform of 2'5'AS was induced following Leydig cells stimulation by the Sendai virus. The most immature germ cells, spermatogonia, were devoid of 2'5'AS whatever the type of stimulation, whereas IFN treatment induced Mx production and increased PKR production in this cell type. IFN stimulation strongly increased PKR production in all three cell types. This new set of data extends our previous investigations and demonstrates that the testis possesses an anti-viral defense system involving IFNs and IFN-induced anti-viral proteins. PMID- 12892904 TI - Expression of growth-regulated oncogene beta in an endometrial epithelial cell line, HHUA, and cultured human endometrial cells. AB - It has been demonstrated that human endometrial epithelial cells (EEC) and stromal cells (ESC) produce a variety of chemokines in vivo and in vitro. Growth regulated oncogene (GRO)beta, which belongs to the CXC chemokine family, is a potent chemoattractant for neutrophils. To evaluate the regulation of GRO beta expression in the endometrium, the production of GRO beta by an EEC line, HHUA, and cultured ESC stimulated with various inflammatory mediators was examined by using an enzyme-linked immunosorbent assay. Unstimulated HHUA and ESC constitutively secreted GRO beta. Interleukin-1 beta, tumor necrosis factor-alpha and interferon-gamma significantly stimulated the expression of GRO beta by HHUA and ESC. Lipopolysaccharide also stimulated the expression of GRO beta by ESC, but not by HHUA. It is suggested that, in the human endometrium, the regulation of GRO beta expression is distinct from that of other CXC chemokines expressed in the endometrium, such as GRO alpha and interleukin-8. The modulation of the GRO beta concentration in the endometrium by inflammatory mediators may contribute to the normal and pathological processes of human reproduction by regulating the trafficking of neutrophils into the endometrium. PMID- 12892905 TI - Identification of photocatalytic degradation products of diazinon in TiO2 aqueous suspensions using GC/MS/MS and LC/MS with quadrupole time-of-flight mass spectrometry. AB - The photocatalytic degradation of the organophosphorus insecticide diazinon in aqueous suspensions has been studied by using titanium dioxide as a photocatalyst. The degradation of the insecticide was a fast process and included the formation of several intermediates that were identified using GC/ion-trap mass spectrometry with EI or CI in positive and negative ionization mode and HPLC/electrospray-QqTOF mass spectrometry. Since primarily hydroxy derivatives were identified in these aqueous suspensions, the mechanism of degradation was probably based on hydroxyl radical attack. The initial oxidative pathways of the degradation of diazinon involved the substitution of sulfur by oxygen on the Pz.dbnd6;S bond, cleavage of the pyrimidine ester bond, and oxidation of the isopropyl group. Exact mass measurements of the derivatives allowed the elemental formula of the molecules to be determined confidently. Similarities to the metabolic pathways occurring in living organisms were observed. PMID- 12892906 TI - Analytical approach for description of ion motion in quadrupole mass spectrometer. AB - Implementation of the analytical method of the solution of the Mathieu equation in conjunction with the algebraic presentation of Mathieu functions is discussed in this work. This approach is used for the analytical expression of fundamental properties of the quadrupole field such as ion trajectory stability and transmission. Extensive comparison with the matrix method is presented with demonstration of the fundamental advantages of the analytical method. However, contrary to the matrix method, the analytical method is limited to the cos trapping waveforms. PMID- 12892907 TI - Sequence confirmation of modified oligonucleotides using IRMPD in the external ion reservoir of an electrospray ionization Fourier transform ion cyclotron mass spectrometer. AB - Modified oligonucleotides continue to play an important role as antisense compounds that inhibit the expression of genes associated with metabolic disorders, cancer, and infectious diseases. Because the majority of modifications render these molecules refractory to standard enzymatic sequencing techniques, alternative sequencing methods which are fast and reliable are needed. In this work we explore how sugar and backbone modifications affect fragmentation patterns observed from oligonucleotides which are fragmented by infrared multiple photon dissociation in the external reservoir of an electrospray ionization Fourier transform ion cyclotron mass spectrometer. The modifications influence which fragment types (i.e., a(n)-B versus c(n)) dominate and the ease with which the oligonucleotides are fragmented. General observations for confirming the sequence of oligonucleotides are described. PMID- 12892908 TI - MS2Assign, automated assignment and nomenclature of tandem mass spectra of chemically crosslinked peptides. AB - In a previous report (Young et al., Proc. Natl. Acad. Sci. U.S.A. 2000, 97, 5802 5806), we provided a proof-of-principle for fold recognition of proteins using a homobifunctional amine-specific chemical crosslinking reagent in combination with mass spectrometry analysis and homology modeling. In this current work, we propose a systematic nomenclature to describe the types of peptides that are generated after proteolysis of crosslinked proteins, their fragmentation by tandem mass spectrometry, and an automated algorithm for MS/MS spectral assignment called "MS2Assign." Several examples are provided from crosslinked peptides and proteins including HIV-integrase, cytochrome c, ribonuclease A, myoglobin, cytidine 5-monophosphate N-acetylneuraminic acid synthetase, and the peptide thymopentin. Tandem mass spectra were obtained from various crosslinked peptides using post source decay MALDI-TOF and collision induced dissociation on a quadrupole-TOF instrument, along with their automated interpretation using MS2Assign. A variety of possible outcomes are described and categorized according to the number of modified lysines and/or peptide chains involved, as well as the presence of singly modified (dead-end) lysine residues. In addition, the proteolysis and chromatographic conditions necessary for optimized crosslinked peptide recovery are presented. PMID- 12892909 TI - Application of self-ionization for enhancing stereochemical and positional effects from arylglycosides under electron ionization conditions in an ion trap mass spectrometer. AB - Ion trap mass spectrometry has been used to structurally characterize and differentiate positional and stereo isomers of arylglycosides having potential antioxidant properties. The use of the self-ionization (SI) technique has allowed to evidence a strong reactivity of fragment ions produced from dissociations of the molecular ion towards the molecules introduced into the trap. Specific structural effects due to positional isomers and anomers have been also envisaged through the occurrence of bimolecular processes inside the ion trap analyzer. Under self-ionization conditions, even-electron ions are produced. The charge is retained on the sugar moiety, in agreement with its proton affinity higher than that of the substituted phenol moiety. Most of the fragmentation pathways involve elimination of acetic acid that protects the hydroxylic groups of the glycoside. SI also produces adduct ions, likely as covalent species, having higher m/z values than the molecular ion. The reaction site is mainly the double bond present in the pyranosidic ring. Even if some fragment ions have lost the initial stereochemistry, their formation can be related to the structure of the parent neutrals introduced into the cell. Collision-induced dissociation (CID) experiments, carried out on ions formed by ion-molecule reactions, have allowed to obtain further information on gas phase ion structures. The study of mass selected ion-molecule reactions and their kinetics have evidenced a spectacularly different reactivity of the ion at m/z 111 towards the two anomers 2alpha and 2beta, with the latter showing a much more pronounced reactivity. The approach developed in this work revealed to be an useful tool in structural characterization, as well as in stereo and regiochemical differentiation of arylglycosides. PMID- 12892910 TI - Separation and identification of DMPO adducts of oxygen-centered radicals formed from organic hydroperoxides by HPLC-ESR, ESI-MS and MS/MS. AB - Many electron spin resonance (ESR) spectra of 5,5-dimethyl-1-pyrroline N-oxide (DMPO) radical adducts from the reaction of organic hydroperoxides with heme proteins or Fe(2+) were assigned to the adducts of DMPO with peroxyl, alkoxyl, and alkyl radicals. In particular, the controversial assignment of DMPO/peroxyl radical adducts was based on the close similarity of their ESR spectra to that of the DMPO/superoxide radical adduct in conjunction with their insensitivity to superoxide dismutase, which distinguishes the peroxyl adducts from the DMPO/superoxide adduct. Although recent reports assigned the spectra suggested to be DMPO/peroxyl radical adducts to the DMPO/methoxyl adduct based on independent synthesis of the adduct and/or (17)O-labeling, (17)O-labeling is extremely expensive, and both of these assignments were still based on hyperfine coupling constants, which have not been confirmed by independent techniques. In this study, we have used online high performance liquid chromatography (HPLC or LC)/ESR, electrospray ionization-mass spectrometry (ESI-MS) and tandem mass spectrometry (MS/MS) to separate and directly characterize DMPO oxygen-centered radical adducts formed from the reaction of Fe(2+) with t-butyl or cumene hydroperoxide. In each reaction system, two DMPO oxygen-centered radical adducts were separated and detected by online LC/ESR. The first DMPO radical adduct from both systems showed identical chromatographic retention times (t(R) = 9.6 min) and hyperfine coupling constants (a(N) = 14.51 G, a(H)(beta) = 10.71 G, and a(H)(gamma) = 1.32 G). The ESI-MS and MS/MS spectra demonstrated that this radical was the DMPO/methoxyl radical adduct, not the peroxyl radical adduct as was thought at one time, although its ESR spectrum is nearly identical to that of the DMPO/superoxide radical adduct. Similarly, based on their MS/MS spectra, we verified that the second adducts (a(N) = 14.86 G and a(H)(beta) = 16.06 G in the reaction system containing t-butyl hydroperoxide and a(N) = 14.60 G and a(H)(beta) = 15.61 G in the reaction mixture containing cumene hydroperoxide), previously assigned as DMPO adducts of t-butyloxyl and cumyloxyl radical, were indeed from trapping t-butyloxyl and cumyloxyl radicals, respectively. PMID- 12892911 TI - Analysis of unsaturated compounds by Ag+ coordination ionspray mass spectrometry: studies of the formation of the Ag+/lipid complex. AB - Coordination ionspray mass spectrometry (CIS-MS) is a useful tool in the detection and identification of cholesterol ester and phospholipid hydroperoxides and diacyl peroxides. Extensive studies of a series of cholesterol esters using CIS-MS revealed the following: (1) Cholesterol esters with equal number of double bonds as the internal standard showed a linear relative response in the mass spectrometer while compounds with non-equal numbers of double bonds gave a nonlinear relative response. (2) Complex adducts containing cholesterol ester, silver ion, AgF, AgBF(4), and 2-propanoxide form when silver is in molar excess of cholesterol esters, reducing the [M + Ag](+) signal. (3) In a mixture of cholesterol esters where silver is limiting, Ch22:6 and Ch20:4 bind to silver at the expense of Ch18:2 and have a higher signal in the mass spectrometer. (4) In a mixture of cholesterol esters where silver concentration is twofold greater than total cholesterol ester concentration, Ch22:6 and Ch20:4 form large complex adducts more frequently than Ch18:2 and have a lower signal in the mass spectrometer. PMID- 12892913 TI - Irradiation effects in MALDI, ablation, ion production, and surface modifications. Part II. 2,5-dihydroxybenzoic acid monocrystals. AB - Irradiation effects at low and high laser fluence on 2,5-dihydroxybenzoic acid large crystals were investigated. Contrary to what was observed for matrices as cinnamic acid derivatives, no chemical degradation of matrix is evidenced and continuous ablation as well as ion production resulted of extended irradiation in all the fluence range corresponding to classical matrix-assisted laser desorption /ionization. Ripples are formed on the base of the crater for a limited number of laser shots under moderate fluence. For extended irradiation, conical shape craters are formed with the axis of the crater oriented along the incident direction of the laser beam. A study of the craters showed that ablation through the ablated volume slowly varied with the laser fluence when a strong increase of ion production (matrix and analyte) was recorded. Ablation volume was found to vary non-linearly with the number of laser shots. On a same spot, the ablated volume and the ion production were measured as a function of the laser energy. With an increasing laser energy (or fluence), the ablated volume slowly increases when the ion production strongly increases. This gives evidence of a decoupling between ablation and ionization. Interaction of the plume with the incoming beam is thus probable. PMID- 12892912 TI - A mass spectrometry study of tirapazamine and its metabolites. insights into the mechanism of metabolic transformations and the characterization of reaction intermediates. AB - Tandem mass spectrometry methods were used to study the sites of protonation and for identification of 3-amino-1,2,4-benzotriazine 1,4-dioxide (1, tirapazamine), and its metabolites (3-amino-1,2,4-benzotriazine 1-oxide (3), 3-amino-1,2,4 benzotriazine 4-oxide (4), 3-amino-1,2,4-benzotriazine (5), and a related isomer 3-amino-1,2,4-benzotriazine 2-oxide (6). Fragmentation pathways of 3 and 5 indicated the 4-N-atom as the most likely site of protonation. Among the N-oxides studied, the 4-oxide (4) showed the highest degree of protonation at the oxygen atom. The differences in collision-induced dissociation of isomeric protonated 1 , 2- and 4-oxides allowed for their identification by LC/MS/MS. Gas phase and liquid phase protonation of tirapazamine occurred exclusively at the oxygen in the 4-position. A loss of OH radical from these ions (2(+)) resulted in ionized 3. Neutralization-reionization mass spectrometry (NR MS) experiments demonstrated the stability of the neutral analogue of protonated tirapazamine in the gas phase in the micro s time-frame. A significant portion of the neutral tirapazamine radicals (2) dissociated by loss of hydroxyl radical during the NR MS event, which indicates that previously proposed mechanisms for redox-activated DNA damage are reasonable. The activation energy for loss of hydroxyl radical from activated tirapazamine (2) was estimated to be approximately 14 kcal mol(-1). Stable neutral analogues of [3 + H](+) and [5 + H](+) ions were also generated in the course of NR MS experiments. Structures of these radicals were assigned to the molecules having an extra hydrogen atom at one of the ring N-atoms. Quantum chemical calculations of protonated 1, 3, 4 and 5 and the corresponding neutrals were performed to assist in the interpretation of experimental results and to help identify their structures. PMID- 12892914 TI - Termolecular ion-molecule reactions in Titan's atmosphere. IV. A search made at up to 1 micron in pure hydrocarbons. AB - The results of a study of ion-molecule reactions occurring in pure methane, acetylene, ethylene, ethane, propyne, propene, propane, and diacetylene at pressures up to 40 microns of pressure are reported. A variety of experimental methods are used: The standard double resonance in an ICR, for determination of the precursor ions and the modulated double resonance ejection in an ICR, for the determination of the daughter ions. The FA-SIFT technique was used for validation and examination of termolecular reactions with rate coefficients that are less than 10(-26) cm(6) s(-1). An extensive database of reaction kinetics already exists for many of these reactions. The main point of this study was the determination of the accuracy of this database and to search for any missing reactions and reaction channels that may have been omitted from earlier investigations. A specific objective of this work was to extend the study to the highest pressures possible to find out if there were any important termolecular reaction channels occurring. A new approach was used here. In the pure hydrocarbon gases the mass spectra were followed as a function of the pressure changes of the gas. An initial guess was first made using the current literature as a source of the reaction kinetics that were expected. A model of the ion abundances was produced from the solution of the partial differential equations in terms of reaction rate coefficients and initial abundances. The experimental data was fitted to the model for all of the pressures by a least squares minimization to the reaction rate coefficients and initial abundances. The reaction rate coefficients obtained from the model were then compared to the literature values. Several new channels and reactions were discovered when the modeled fits were compared to the actual data. This is all explained in the text and the implications of these results are discussed for the Titan atmosphere. PMID- 12892915 TI - Investigation of ion/molecule reactions as a quantification method for phosphorylated positional isomers. an FT-ICR approach. AB - A rapid and accurate method of quantifying positional isomeric mixtures of phosphorylated hexose and N-acetylhexosamine monosacchrides by using gas-phase ion/molecule reactions coupled with FT-ICR mass spectrometry is described. Trimethyl borate, the reagent gas, reacts readily with the singly charged negative ions of phosphorylated monosaccharides to form two stable product ions corresponding to the loss of one or two neutral molecules of methanol from the original adduct. Product distribution in the ion/molecule reaction spectra differs significantly for isomers phosphorylated in either the 1- or the 6 position. As a result, the percents of total ion current of these product ions for a mixture of the two isomers vary with its composition. In order to determine the percentage of each isomer in an unknown mixture, a multicomponent quantification method is utilized in which the percents of total ion current of the two product ions for each pure monosaccharide phosphate and the mixture are used in a two-equation, two-unknown system. The applicability of this method is demonstrated by successfully quantifying mock mixtures of four different isomeric pairs: Glucose-1-phosphate and glucose-6-phosphate; mannose-1-phosphate and mannose-6-phosphate; galactose-1-phosphate and galactose-6-phosphate; N acetylglucosamine-1-phosphate and N-acetylglucosamine-6-phosphate. The effects of mixture concentrations and ion/molecule reaction conditions on the quantification are also discussed. Our results demonstrate that this assay is a fast, sensitive, and robust method to quantify isomeric mixtures of phosphorylated monosaccharides. PMID- 12892917 TI - Proceedings of the 23rd International Specialized Symposium on Yeasts (ISSY-23). August 26-29, 2003. Budapest, Hungary. PMID- 12892918 TI - Adaptive evolution of wine yeast. AB - Alcoholic fermentation is one of the main phases in wine production. It is usually conducted by yeasts belonging to the species Saccharomyces cerevisiae. Industrial S. cerevisiae strains are highly specialized organisms, which have evolved to utilize to their full potential the different environments or ecological niches. So, during the alcoholic fermentation, the yeast has been adapted to different kinds of stress conditions; this adaptation is call "domestication". In this review, we describe the different mechanisms involved in the adaptive evolution of wine yeast strains. PMID- 12892919 TI - Yeast interactions and wine flavour. AB - Wine is the product of complex interactions between fungi, yeasts and bacteria that commence in the vineyard and continue throughout the fermentation process until packaging. Although grape cultivar and cultivation provide the foundations of wine flavour, microorganisms, especially yeasts, impact on the subtlety and individuality of the flavour response. Consequently, it is important to identify and understand the ecological interactions that occur between the different microbial groups, species and strains. These interactions encompass yeast-yeast, yeast-filamentous fungi and yeast-bacteria responses. The surface of healthy grapes has a predominance of Aureobasidium pullulans, Metschnikowia, Hanseniaspora (Kloeckera), Cryptococcus and Rhodotorula species depending on stage of maturity. This microflora moderates the growth of spoilage and mycotoxigenic fungi on grapes, the species and strains of yeasts that contribute to alcoholic fermentation, and the bacteria that contribute to malolactic fermentation. Damaged grapes have increased populations of lactic and acetic acid bacteria that impact on yeasts during alcoholic fermentation. Alcoholic fermentation is characterised by the successional growth of various yeast species and strains, where yeast-yeast interactions determine the ecology. Through yeast bacterial interactions, this ecology can determine progression of the malolactic fermentation, and potential growth of spoilage bacteria in the final product. The mechanisms by which one species/strain impacts on another in grape-wine ecosystems include: production of lytic enzymes, ethanol, sulphur dioxide and killer toxin/bacteriocin like peptides; nutrient depletion including removal of oxygen, and production of carbon dioxide; and release of cell autolytic components. Cell-cell communication through quorum sensing molecules needs investigation. PMID- 12892920 TI - Spoilage yeasts in the wine industry. AB - Yeasts play a central role in the spoilage of foods and beverages, mainly those with high acidity and reduced water activity (a(w)). A few species are capable of spoiling foods produced according to good manufacturing practices (GMPs). These can survive and grow under stress conditions where other microorganisms are not competitive. However, many of the aspects determining yeast spoilage have yet to be clarified. This critical review uses the wine industry as a case study where serious microbiological problems are caused by yeasts. First, the limitations of the available tools to assess the presence of spoilage yeasts in foods are discussed. Next, yeasts and factors promoting their colonisation in grapes and wines are discussed from the ecological perspective, demonstrating that a deeper knowledge of vineyard and winery ecosystems is essential to establish the origin of wine spoilage yeasts, their routes of contamination, critical points of yeast infection, and of course, their control. Further, zymological indicators are discussed as important tools to assess the microbiological quality of wines, although they are rarely used by the wine industry. The concepts of the susceptibility of wine to spoilage yeasts and wine stability are addressed based on scientific knowledge and industrial practices for monitoring yeast contamination. A discussion on acceptable levels of yeasts and microbiological criteria in the wine industry is supported by data obtained from wineries, wholesalers, and the scientific literature.Finally, future directions for applied research are proposed, involving collaboration between scientists and industry to improve the quality of wine and methods for monitoring the presence of yeast. PMID- 12892921 TI - The role of interaction between yeasts and lactic acid bacteria in African fermented milks: a review. AB - Yeasts are present in indigenous African fermented milks in numbers up to log 8 cfu g(-1), together with a varied lactic acid bacteria (LAB) flora, and therefore potentially contribute to product characteristics. However, interaction between yeasts and LAB in these products has received little notice. In studies of indigenous fermented milk in Zimbabwe and Uganda, many samples contained more than one species of yeast, but Saccharomyces cerevisiae was most commonly isolated. Other frequent isolates were other species of Saccharomyces and several species of Candida. Most yeast isolates were lactose-negative but usually galactose-positive. Some strains assimilated lactate and citrate. The growth in milk of strains of yeasts and LAB, isolated from naturally soured milk in Zimbabwe, and their interaction when selected pairs of strains were grown together has been studied. Interactions were shown by the significantly different amounts of certain metabolites produced, such as acetaldehyde and malty aldehydes, when co-cultures were compared to pure cultures. Preliminary sensory acceptance tests did not show, however, that milks made from a co-culture with Candida kefyr and LAB were preferable to the pure LAB culture. Further work is still needed to elucidate the reactions that may be taking place in fermented milk between varying LAB and yeast populations. The potential for use as starter cultures depends on various aspects, including the final product being prepared. The role of other microorganisms in naturally fermented milk also needs to be studied. PMID- 12892922 TI - Evaluation of ribosomal RNA and actin gene sequences for the identification of ascomycetous yeasts. AB - Highly similar gene sequences of the 5' region of the large subunit (LSU) are commonly interpreted to predict the organism's identity. However, it was recognised that closely related taxa do not always show sufficiently diverged D1/D2 LSU sequences to differentiate between them. The effectiveness of species separation using D1/D2 LSU sequences, small subunit (SSU) sequences and actin gene sequences was determined by pair-wise comparisons. The LSU data showed coinciding similarities among and within species. The actin data resolved all investigated species. Examples strengthened the value of almost complete SSU sequences for species separation. The larger number of differences in the highly conserved actin gene, compared to the overall more variable LSU gene, is due to the tolerance of protein coding genes to synonymous nucleotide changes. In contrast, the pairing in secondary structures of the rRNA, ensuring the functionality of the molecule, relies on longer and uninterrupted sequence sections. In conclusion, D1/D2 LSU sequences are not specific enough to identify closely related taxa. The actin gene is a better marker in these cases. However, because of the availability of a large database of fungal D1/D2 LSU sequences, this gene region is currently still the preferred target for sequence-based identification. PMID- 12892923 TI - Ura- host strains for genetic manipulation and heterologous expression of Torulaspora delbrueckii. AB - Recently, the industrial and academic interest in the yeast Torulaspora delbrueckii has increased notably due to its high resistance to several stresses. This characteristic has made of this organism a very attractive model to study the molecular basis of the stress response in yeast. However, very little is known about the physiology and genetics of this yeast, and the tools for its manipulation have not been developed. Here, we have generated Ura(-) strains of the baker's yeast T. delbrueckii IGC5323 by either 5-FOA-aided selection or transformation with a PCR-based disruption cassette, natMX4, which confers nourseothricin resistance. Furthermore, the mutant and disruptant strains were used as recipient of a plasmid containing the xlnB cDNA from Aspergillus nidulans. Our results indicate that Torulaspora transformants produce active recombinant protein at a similar level to that found for Saccharomyces. PMID- 12892924 TI - The microbial ecology of cocoa bean fermentations in Indonesia. AB - Cocoa beans are the principal raw material of chocolate manufacture. The beans are subject to a microbial fermentation as the first stage in chocolate production. The microbial ecology of bean fermentation (Forastero and Trinitario cultivars) was investigated at three commercial fermentaries in East Java, Indonesia by determining the populations of individual species at 12-h intervals throughout the process. The first 2-3 days of fermentation were characterised by the successional growth of various species of filamentous fungi, yeasts, lactic acid bacteria and acetic acid bacteria. The principal species found were Penicillium citrinum, an unidentified basidiomycete, Kloeckera apis, Saccharomyces cerevisiae, Candida tropicalis, Lactobacillus cellobiosus, Lactobacillus plantarum and Acetobacter pasteurianus. The later stages of fermentation were dominated by the presence of Bacillus species, mostly, Bacillus pumilus and Bacillus licheniformis. Glucose, fructose, sucrose and citric acid of the bean pulp were utilised during fermentation, with the production of ethanol, acetic acid and lactic acid that diffused into the beans. The filamentous fungi were notable for their production of polygalacturonase activity and probably contributed to the degradation of bean pulp. PMID- 12892925 TI - Proteolytic yeasts isolated from raw, ripe tomatoes and metabiotic association of Geotrichum candidum with Salmonella. AB - Metabiotic associations between food-borne fungi and bacteria capable of causing human diseases are a public health concern. A survey of decayed and damaged, uncooked, ripe tomatoes was done to determine the presence and prevalence of yeasts capable of increasing the pH pulp tissue, thus creating a more favorable environment for survival and growth of enteric pathogens. Sixty-two of the 371 (16.7%) fungi isolated from 215 decayed or damaged tomatoes; 12 of the 62 (19.4%) yeasts showed proteolytic activity on gelatin agar (GA) and/or standard methods caseinate (SMC) agar. The pH of tomato pericarp (pulp) tissue from which 9 of the 12 yeasts were isolated ranged from 4.3 to 7.5 (mean=5.3) compared to 4.2-5.1 (mean=4.8) for sound pulp tissue in the same tomatoes. The 12 proteolytic yeasts consisted of four strains of Cryptococcus albidus, two strains each of Debaryomyces hansenii and Trichosporon pullulans, and one strain each of Cryptococcus humicolus, Cryptococcus laurentii, Geotrichum candidum, and Sporidiobolus pararoseus. Survival and growth characteristics of a five-serotype mixture of Salmonella co-inoculated with G. candidum into sound (not chill injured) and chill-injured tomatoes were studied. Storage of sound tomatoes at 15 degrees C for 10 days resulted in an increase in population of 7.6 log(10) cfu of Salmonella/g of a 2-g sample of co-infected pulp tissue. Increases were less in tissue inoculated with Salmonella only, Salmonella on day 0 followed by G. candidum on day 3, or G. candidum on day 0 followed by Salmonella on day 3. Trends were similar in sound inoculated tomatoes stored at 25 degrees C. Growth of Salmonella was enhanced in chill-injured tomatoes compared to sound tomatoes; a population of 10 log(10) cfu/g of chill-injured pulp tissue was reached within 10 days at 25 degrees C. Results clearly show that growth of a proteolytic, alkalinizing yeast such as G. candidum in raw tomatoes enhances conditions for growth of Salmonella. The removal of tomatoes infected with proteolytic yeasts and other fungi from lots intended for minimally processed tomato products is an essential step in reducing the risk of human diseases caused by pathogenic bacteria favored by increased pH of decayed pulp tissue. PMID- 12892926 TI - Analysis of the inhibition of food spoilage yeasts by vanillin. AB - The antimicrobial potential of vanillin, the major component of vanilla flavour, was examined against the growth of three yeasts associated with food spoilage, Saccharomyces cerevisiae, Zygosaccharomyces bailii and Zygosaccharomyces rouxii. Minimum inhibitory concentration (MIC) values of 21, 20 and 13 mM vanillin were determined for the three yeast strains, respectively. The observed inhibition was found to be biostatic. During fermentation, the bioconversion of sub-MIC levels of vanillin in the culture medium was demonstrated. The major bioconversion product was identified as vanillyl alcohol, however low levels of vanillic acid were also detected. Neither the vanillyl alcohol nor the vanillic acid was found to be antagonistic to yeast cell growth. The results indicate the importance of the aldehyde moiety in the vanillin structure regarding its antimicrobial activity and that the bioconversion of vanillin could be advantageous for the yeasts, but only at levels below MIC. These bioconversion activities, presumably catalysed by non-specific dehydrogenases, were shown to be expressed constitutively. It was observed that increased vanillin concentrations inhibited its own bioconversion suggesting that the activity required intact cells with metabolic capacity. PMID- 12892927 TI - Characterization of yeast isolates originating from Hungarian dairy products using traditional and molecular identification techniques. AB - Yeast isolates collected from various Hungarian dairy products were identified using simplified identification system (SIM) and restriction fragment analysis of PCR-amplified 18S rDNA with the neighbouring ITS1 region (ITS-PCR; ribotyping). The isolates were grouped into 26 species; Geotrichum candidum, Debaryomyces hansenii, Yarrowia lipolytica, Kluyveromyces lactis and Candida catenulata were found as the predominant species. SIM and ITS-PCR proved to be useful and convenient taxonomic tools for rapid identification at species level. Two most frequent species, G. candidum and D. hansenii, were further characterized by randomly amplified polymorphic DNA (RAPD-PCR) analysis. RAPD-PCR using M13 primer resulted in discrimination between most strains of the same species and allowed a certain degree of intraspecific typing. PMID- 12892928 TI - Yeasts as adjunct starters in matured Cheddar cheese. AB - Debaryomyces hansenii and Yarrowia lipolytica are typical foodborne yeast species frequently associated with dairy products and capable of predominating the yeast composition in such systems. The two species fulfil a number of criteria to be regarded as co-starters for cheesemaking. They are known for their proteolytic and lipolytic activity as well as their compatibility and stimulating action with the lactic acid starter cultures when co-inoculated. Recent studies indicated that yeasts could be included as part of starter cultures for the manufacturing of cheese, enhancing flavour development during the maturation. The potential of D. hansenii and Y. lipolytica as agents for accelerated ripening of matured Cheddar cheese has been evaluated during four cheese treatments. The interaction between the two yeast species and the lactic acid bacteria was surveyed incorporating (i) D. hansenii, (ii) Y. lipolytica, (iii) both species as adjuncts to the starter culture and (iv) a control cheese without any additions for the production of matured Cheddar cheese. The physical and chemical properties of the cheeses were monitored in order to evaluate the contribution of the yeasts to cheese maturation. The yeasts grew in association with the lactic acid bacteria without any inhibition. The yeasts species when individually added contributed to the development of bitter flavours despite accelerated development of strong Cheddar flavours. When both species were incorporated as part of the starter culture, the cheese, however, had a good strong flavour after a reduced ripening period. The cheese retained this good flavour and aroma after 9 months of production. The simultaneous application of D. hansenii and Y. lipolytica as part of the starter culture for the production of matured Cheddar cheese is proposed. PMID- 12892929 TI - Purification and properties of an arginyl aminopeptidase from Debaryomyces hansenii. AB - A metallo arginyl aminopeptidase (EC 3.4.11.6) activated by Co(2+) was isolated from Debaryomyces hansenii CECT 12487. The enzyme was purified after precipitation with protamine sulphate, followed by a weak anion exchange chromatography, gel filtration chromatography and a strong anion exchange chromatography. The arginyl aminopeptidase (AAP) was purified 337 folds, with a 18% recovery. The AAP appeared to be a dimer with a molecular mass of 101 kDa. The enzyme was active in the pH range from 6 to 9. The optimal activity was detected at pH 7.0 and at 37 degrees C. AAP activity was inhibited by typical aminopeptidase inhibitors (puromycin and bestatin), reducing agents (DTT), chelating agents (EDTA, EGTA and phenantroline) and sulphydryl groups reagents (iodoacetate). Ca(2+), Mn(2+) and Co(2+) activated the enzyme, while Cu(2+), Cd(2+), Hg(2+) and Mg(2+) inhibited it. The K(m) values calculated for Arg-AMC (7 amido-4-methylcoumarin) and Leu-AMC were 0.071 and 0.094 mM, respectively. The enzyme showed maximum specificity for basic amino acids (Arg and Lys), but was also able to hydrolyze non-charged amino acids (Leu, Met and Ala) and, at a minor rate, aromatic amino acids (Phe and Tyr). AAP showed higher activity when an acid residue was located at the C-terminal position of dipeptides. The described purification of an arginyl aminopeptidase from the yeast D. hansenii can contribute to the lack of knowledge about the exopeptidase activity in one of the yeasts more frequently isolated in sausage and to understand its role during the ripening of a fermented sausage. PMID- 12892930 TI - Changes in wine yeast storage carbohydrate levels during preadaptation, rehydration and low temperature fermentations. AB - The metabolism of glycogen and trehalose was analysed in a wine yeast strain fermenting at 25 and 13 degrees C. Trehalose and glycogen degradation were completed during the lag phase of fermentation. Ammonia was taken up rapidly and once it had been reduced to negligible amounts, the synthesis of trehalose started. Glycogen followed a similar pattern. If trehalose synthesis was taken as a stress indicator, the fermentation at 13 degrees C could not be considered stressful because the maximum concentrations are similar at both temperatures. In industrial fermentations, and after a preadaptation in grape must for several hours at 18 degrees C, the lag phase was reduced significantly, and this may be why trehalose and glycogen were completely depleted at the beginning of the low temperature fermentation. Various preadaptation conditions were tested so that their influence on trehalose and glycogen degradation could be determined. The presence of fermentable carbon sources, such as glucose or fructose, triggered the mobilisation and use of trehalose. However, just increasing the osmotic pressure did not reduce the trehalose content. No such differences were observed in glycogen metabolism. PMID- 12892931 TI - The species-specific ratios of 2,3-butanediol and acetoin isomers as a tool to evaluate wine yeast performance. AB - The isomers of 2,3-butanediol [R,R; S,S; R,S (meso-form)] and of acetoin (R,S) were determined in laboratory wine fermentations carried out by 50 yeast strains, 10 for each of the following species, Saccharomyces cerevisiae, Kloeckera apiculata, Candida stellata, Metschnikowia pulcherrima and Brettanomyces bruxellensis, in order to evaluate if such parameters might be used to differentiate wines obtained with different yeast species. According to analysis of variance (ANOVA), the strains of the same species behaved similarly, whereas the five yeast species behaved differently so that species-specific profiles were recognized. Moreover, the discriminant analysis grouped the wines into five groups, each including the 10 wines obtained by the 10 yeast strains of the same species. Trials were also included where musts partially fermented by non Saccharomyces species were inoculated with a selected strain of S. cerevisiae to complete fermentation, and the content in 2,3-butanediol and acetoin isomers was again determined and statistical analysis was performed. Although the final values of these parameters resembled those obtained in pure fermentation with S. cerevisiae, statistical analysis discriminated wines according to the yeast species performing the first fermentation phase. PMID- 12892932 TI - Function of yeast species and strains in wine flavour. AB - The diversity and the composition of the yeast micropopulation significantly contribute to the sensory characteristics of wine. The growth of each wine yeast species is characterized by a specific metabolic activity, which determines concentrations of flavour compounds in the final wine. However, it must be underlined that, within each species, significant strain variability has been recorded. The wide use of starter cultures, mainly applied to reduce the risk of spoilage and unpredictable changes of wine flavour, can ensure a balanced wine flavour, but it may also cause a loss of characteristic aroma and flavour determinants. Thus, the beneficial contribution from the yeast increases when starter cultures for winemaking are selected on the basis of scientifically verified characteristics and are able to complement and optimise grape quality and individual characteristics. Here we report the characterization of a large number of strains of different wine yeast species, isolated from spontaneous wine fermentations and included in the culture collection of the Basilicata University. PMID- 12892933 TI - Acetate ester formation in wine by mixed cultures in laboratory fermentations. AB - Two non-Saccharomyces wine yeast strains, Hanseniaspora guilliermondii 11104 and Pichia anomala 10590, selected as good producers of acetate esters when grown on synthetic microbiological medium, have been tested in wine fermentations as mixed cultures together with Saccharomyces cerevisiae. Wines produced using mixed cultures showed levels of acetaldehyde, acetic acid, glycerol and total higher alcohols within the ranges described for wine, whereas an increase in acetate ester concentrations was found. Ethyl acetate was the main ester produced, and isoamyl acetate and 2-phenylethyl acetate made up the next largest group of ester compounds in the wines analysed. H. guilliermondii 11104 was found to be a strong producer of 2-phenylethyl acetate in both pure and mixed cultures whereas S. cerevisiae was the best producer of ethyl esters. Mixed cultures did not influence ethyl ester levels at all. PMID- 12892934 TI - CHROMagar Candida medium as a practical tool for the differentiation and presumptive identification of yeast species isolated from salads. AB - CHROMagar Candida medium was used to study the diversity of yeast biota of salad samples, and to presumptively identify the isolates. This medium was originally developed for the selective isolation and presumptive identification of some clinically important yeast species such as Candida albicans, Candida tropicalis, Candida krusei, and Candida glabrata on the basis of differences in colour and surface of colonies. Ninety three yeast strains representing 33 species from the culture collection and 39 fresh isolates from different mayonnaise-based mixed salads showed a wide range of hue of colony colours ranging from white to yellow, orange, red, pink, purple, blue, green, etc., as well as different morphological appearances on the CHROMagar Candida medium. Therefore, CHROMagar Candida medium facilitates the detection of mixtures of yeast species from different samples on a single isolation plate and this medium can be a practical method for the differentiation and rapid presumptive identification of many yeast species occurring frequently in different kind of foods. PMID- 12892935 TI - Yeast associated with spontaneous fermentations of white wines from the "Txakoli de Bizkaia" region (Basque Country, North Spain). AB - The microbiota of eight spontaneous fermentation of white wine from different grape varieties and different wineries from the "Txakoli de Bizkaia" region (Basque country, North Spain), in 1996 and 1997 campaigns was studied. The yeast population was higher in grapes harvested in 1997, in which late summer and early autumn was warmer and drier. Eight species belonging to five genera were identified in total. The most frequent genera in grapes were Rhodotorula in 1996 and Kloeckera in 1997. Saccharomyces bayanus was the most frequent species during vigorous and final fermentation, and it was occasionally isolated from grapes and must. Only another Saccharomyces spp., i.e., S. kluyvery, was identified in some samples from 1997. PMID- 12892937 TI - Synthesis of a mannose nonasaccharide existing in the exopolysaccharide of Cryphonectria parasitica. AB - alpha-D-Manp-(1-->2)-alpha-D-Manp-(1-->2)-alpha-D-Manp-(1-->6)[alpha-D-Manp-(1- >3)-alpha-D-Manp-(1-->2)-alpha-D-Manp-(1-->2)]-alpha-D-Manp-(1-->6)-[alpha-D-Manp (1-->2)]-alpha-D-Manp, existing in the exopolysaccharide of Cryphonectria parasitica was synthesized as its allyl glycoside in a regio- and stereoselective manner. PMID- 12892938 TI - Synthesis of a hexasaccharide, the repeating unit of O-deacetylated GXM of C. neoformans serotype A. AB - beta-D-GlcpA-(1-->2)-alpha-D-Manp-(1-->3)-[beta-D-Xylp-(1-->2)]-alpha-D-Manp-(1- >3)[-beta-D-Xylp-(1-->2)]-alpha-D-Manp, the repeating unit of the exopolysaccharide from Cryptococcus neoformans serovar A, was synthesized as its allyl glycoside. Thus, 3-O-selective acetylation of allyl 4,6-O-benzylidene-alpha D-mannopyranoside afforded 2, and subsequent glycosylation of 2 with 2,3,4-tri-O benzoyl-D-xylopyranosyl trichloroacetimidate furnished the beta-(1-->2)-linked disaccharide 4. Debenzylidenation followed by benzoylation gave allyl 2,3,4-tri-O benzoyl-beta-D-xylopyranosyl-(1-->2)-3-O-acetyl-4,6-di-O-benzoyl-alpha-D mannopyranoside (5), and selective 3-O-deacetylation gave the disaccharide acceptor 6. Coupling of 6 with 2-O-acetyl-3,4,6-tri-O-benzoyl-alpha-D mannopyranosyl trichloroacetimidate yielded the trisaccharide 8, and subsequent deallylation and trichloroacetimidation gave 2,3,4-tri-O-benzoyl-beta-D xylopyranosyl-(1-->2)-[2-O-acetyl-3,4,6-tri-O-benzoyl-alpha-D-mannopyranosyl-(1- >3)]-4,6-di-O-benzoyl-alpha-D-mannopyranosyl trichloroacetimidate (9). Condensation of the trisaccharide donor 9 with the disaccharide acceptor 6 gave the pentasaccharide 10 whose 2-O-deacetylation gave the acceptor 11. Glycosylation of 11 with methyl 2,3,4-tri-O-acetyl-alpha-D-glucopyranosyluronate trichloroacetimidate and subsequent deprotection gave the target hexasaccharide. PMID- 12892939 TI - Synthesis of mannose-containing analogues of (1-->6)-branched (1-->3) glucohexaose (I). AB - alpha-D-Manp-(1-->3)-[alpha-D-Manp-(1-->6)]-alpha-D-Glcp-(1-->3)-beta-D-Glcp-(1- >3)-[alpha-D-Manp-(1-->6)]-D-Glcp and alpha-D-Manp-(1-->3)-[beta-D-Glcp-(1-->6)] alpha-D-Glcp-(1-->3)-beta-D-Glcp-(1-->3)[-alpha-D-Manp-(1-->6)]-D-Glcp were synthesized in a regio- and stereoselective way as the mannose-containing analogues of the immunomodulating beta-D-Glcp-(1-->3)-[beta-D-Glcp-(1-->6)]-alpha D-Glcp-(1-->3)-beta-D-Glcp-(1-->3)-[beta-D-Glcp-(1-->6)]-D-Glcp. PMID- 12892940 TI - Novel autoxidation and Michael addition of a butenolide-containing sugar leading to a C-branched-chain glucopyranosidulose, and X-ray structure of intermediates. AB - A butenolide-containing sugar available from the aldol condensation of methyl 4,6 O-benzylidene-alpha-D-glucopyranosid-2-ulose with diethyl malonate is autoxidized at the C-3 position into the corresponding alpha,beta-unsaturated gamma-lactone sugar by air, which subsequently undergoes 1,4-conjugate (Michael) addition of hydroxide ion (or water) leading to a C-branched-chain glucopyranosidulose. The autoxidations are also performed in weakly basic, neutral and weakly acidic medium, respectively. PMID- 12892941 TI - Complexes of sodium vanadate(V) with methyl alpha-D-mannopyranoside, methyl alpha and beta-D-galactopyranoside, and selected O-methyl derivatives: a 51V and 13C NMR study. AB - The hydroxyl group stereochemistry of complexation of sodium vanadate(V) with Me alpha-Manp, Me alpha- and beta-Galp and selected O-methyl derivatives in D(2)O was determined by 51V, 1D and 2D 13C NMR spectroscopy at pD 7.8. The 51V approach served to show the extent of complexation and the minimum number of esters formed. That of Me alpha-Manp gave rise mainly to a 51V signal at delta -515, identical with that of its 4,6-di-O-methyl derivative, which had only a 2,3-cis diol exposed. The 13C NMR spectra contained much weaker signals of the complexes, but both glycosides showed strong C-2 and C-3 alpha-shifts of +17.3 and +10.8 ppm, respectively. As expected, Me 2,3-Me(2)-alpha-Manp, which contains a 4,6 diol, did not complex. Me Galp anomers and their derivatives showed more diversity in the structure of its oxyvanadium derivatives. Me alpha-Galp, with its 3,4-cis-diol, complexed to give rise to 51V signals at delta -495 (9%), -508 (10%), and -534 (4%). These shifts and proportions were maintained with Me beta Galp and Me 6Me-alpha-Galp. 51V NMR spectroscopy showed that Me 3Me-beta-Galp, with its possibly available 4,6-diol, did not complex. Similarly, Me alpha Galp+vanadate gave a 13C DEPT spectrum that did not contain an inverted signal at delta >71.4, as would be expected of a C-6 resonance suffering a strong downfield alpha-shift. Me 2,6-Me(2)-alpha-Galp, with a 3,4-cis-diol group, gave rise to two 51V signals of complexes at delta -492 (9%) and -508 (9%), showing more than one structure of oxyvanadium derivatives. PMID- 12892942 TI - Inclusion complexation behavior of dyestuff guest molecules by a bridged bis(cyclomaltoheptaose)[bis(beta-cyclodextrin)] with a pyromellitic acid diamide tether. AB - A novel bridged bis(beta-cyclodextrin) with a pyromellitic acid 2,5-diamide tether (2) has been synthesized by reaction of 6(I)-(2-aminoethyleneamino)-6 deoxycyclomaltoheptaose [mono 6-(2-aminoethyleneamino)-6-deoxy-beta-cyclodextrin] with 1,2,4,5-benzenetetracarboxylic dianhydride. Its inclusion complexation behavior with some representative dyestuffs, i.e., Acridine Red (AR), Rhodamine B (RhB), Neutral Red (NR), Brilliant Green (BG), was studied by using UV absorption, fluorescence, and 2D NMR spectroscopy. Fluorescence titrations have been performed at 25 degrees C in pH 7.2 buffer solution to calculate the binding constants of resulting complexes. These results obtained indicated that bis(beta cyclodextrin) 2 exhibits the strongly enhanced binding ability with all dye molecules examined compared with natural cyclodextrins. The binding modes of 2 with dye molecules have been deduced by 2D NMR experiments to establish the correlations between molecular conformations and binding constants of inclusion complexation. It is found that the improved binding ability and molecular selectivity of 2 could be attributed to double-cavity cooperative inclusion interaction and the size/shape matching between the host and guest. PMID- 12892943 TI - Biodegradation of poly(epsilon-caprolactone)/starch blends and composites in composting and culture environments: the effect of compatibilization on the inherent biodegradability of the host polymer. AB - The biodegradability of poly(epsilon-caprolactone) (PCL) was studied in blends and composites of modified and granular starch. Four types of PCL-starch compositions were prepared: (i) PCL-granular starch blends; (ii) hydrophobic coating of starch particles by n-butylisocyanate (C(4) starch) and octadecyltrichlorosilane (C(18) starch), followed by melt blending with PCL; (iii) PCL-starch blends compatibilized by PCL-g-dextran grafted copolymer (PGD); and (iv) PCL-grafted starch particles (PGS) as obtained by in situ ring-opening polymerization of caprolactone (CL) initiated directly from hydroxyl functions at the granular starch surface. Biodegradability of these materials was measured by monitoring the percentage of weight loss in composting and the rate of fungal colonization when samples were used as a sole carbon source for fungus (A. niger). Intrinsic viscosity [eta] of host PCL chains was measured after extraction of composted samples in boiled chloroform. SEM was used to study the surface morphology after compost incubation of the samples. The inherent biodegradability of host polymer was enhanced with surface compatibilization during composting for longer incubation. It was observed that the weight loss during composting increased with the decrease in interfacial tension between filler and polymer. In general, it was concluded that inherent biodegradability does not depend very significantly on the concentration of starch in the polyester matrix, but on the compatibilization efficiency. The effect of the PCL fraction in the graft copolymer, when used as compatibilizer, was also studied on the biodegradability of the host polymer. PMID- 12892944 TI - Dilute liquid crystals used to enhance residual dipolar couplings may alter conformational equilibrium in oligosaccharides. AB - The solution structures of a trisaccharide and a pentasaccharide containing the Lewis(x) motif were determined by two independent approaches using either dipolar cross-relaxation (NOE) or residual dipolar coupling (RDC) data. For the latter, one-bond 13C[bond](1)H RDC enhanced by two different mineral liquid crystals were used alone. Home-written programs were employed firstly for measuring accurately the coupling constants in the direct dimension of non-decoupled HSQC experiments, secondly for transforming each RDC data set into geometrical restraints. In this second program, the complete molecular structure was expressed in a unique frame where the alignment tensor is diagonal. Assuming that the pyranose rings are rigid, their relative orientation is defined by optimizing the glycosidic torsion angles. For the trisaccharide, a good agreement was observed between the results of both approaches (NOE and RDC). In contrast, for the pentasaccharide, strong discrepancies appeared, which seem to result from interactions between the pentasaccharide and the mesogens, affecting conformational equilibrium. This observation is of importance, as it reveals that using simultaneously NOE and RDC can be hazardous as the former represent 99% of the molecules free in solution, whereas the latter correspond to less than 1% of the structure bound to the mesogen. PMID- 12892945 TI - Synthesis of a novel N[bond]O-interglycosidic disaccharide. AB - The carbohydrate subunits carrying an N-O-interglycosidic bond play a very important role in the biological activity of the enediyne antibiotics. Condensation of O-(alpha- and beta-D-glucopyranosyl)hydroxylamine (5a and 5b) with the hex-3-ulopyranoside (6) furnished methyl 4,6-O-benzylidene-2,3-dideoxy-3 (2,3,4,6-tetra-O-benzyl-alpha-D-glucopyranosyloxy)imino-alpha- and beta-D-erythro hexopyranoside (7a and 7b). Stereoselective reduction of the Cz.dbnd6;N bond of 7a and 7b with sodium cyanoborohydride resulted in the formation of the required protected N-O-interglycosidic disaccharides (8a and 8b). Finally, catalytic hydrogenation of 8a afforded methyl 2,3-dideoxy-3-(alpha-D glucopyranosyloxy)amino-alpha-D-ribo-hexopyranoside (9a). Under similar conditions the beta anomer 8b underwent decomposition. PMID- 12892946 TI - Synthesis and characterization of a sulfated pentasaccharide containing the Lewisx motif. AB - We report the synthesis of a sulfated pentasaccharide containing the Lewis(x) motif used for an NMR study described in Carbohydr. Res. 2003, 338, this issue, see following communication: doi:10.1016/S0008-6215(03)00243-X, using the dibutylstannylene acetal methodology. PMID- 12892947 TI - Synthetic methyl hexagalacturonate hapten inhibitors of anti-homogalacturonan monoclonal antibodies LM7, JIM5 and JIM7. AB - A range of synthetic methyl hexagalacturonates were used as potential hapten inhibitors in competitive-inhibition enzyme-linked immunosorbent assays (ELISAs) with anti-homogalacturonan monoclonal antibodies LM7, JIM5 and JIM7. The selective inhibition of these antibodies by different haptens provides insight into the structures of the partially methyl-esterified pectin epitopes of these widely used monoclonal antibodies. PMID- 12892948 TI - Structure of the biological repeating unit of the O-antigen of Pseudomonas aeruginosa immunotype 4 containing both 2-acetamido-2,6-dideoxy-D-glucose and 2 acetamido-2,6-dideoxy-D-galactose. AB - A phosphorylated core-lipid A backbone oligosaccharide that carries a disaccharide remainder of the first O-antigen repeating unit was derived by strong alkaline degradation following mild hydrazinolysis of the lipopolysaccharide of Pseudomonas aeruginosa immunotype 4 (serogroup O-1). The structure of the oligosaccharide was determined using ESI MS and NMR spectroscopy and it was demonstrated that 2-acetamido-2,6-dideoxy-D-glucose is the first monosaccharide of the O-polysaccharide that is linked to the LPS core. These data define the structure of the biological repeating unit of the O-antigen. PMID- 12892949 TI - What price competitiveness in the drugs industry? PMID- 12892950 TI - Smoking in movies: a major problem and a real solution. PMID- 12892951 TI - Human genetic variation and disease. PMID- 12892952 TI - TRIPS: whose interests are being served? PMID- 12892953 TI - New biology of the oestrogen receptor. PMID- 12892954 TI - Child survival: a global health challenge. PMID- 12892955 TI - Newly discovered coronavirus as the primary cause of severe acute respiratory syndrome. AB - BACKGROUND: The worldwide outbreak of severe acute respiratory syndrome (SARS) is associated with a newly discovered coronavirus, SARS-associated coronavirus (SARS CoV). We did clinical and experimental studies to assess the role of this virus in the cause of SARS. METHODS: We tested clinical and postmortem samples from 436 SARS patients in six countries for infection with SARS-CoV, human metapneumovirus, and other respiratory pathogens. We infected four cynomolgus macaques (Macaca fascicularis) with SARS-CoV in an attempt to replicate SARS and did necropsies on day 6 after infection. FINDINGS: SARS-CoV infection was diagnosed in 329 (75%) of 436 patients fitting the case definition of SARS; human metapneumovirus was diagnosed in 41 (12%) of 335, and other respiratory pathogens were diagnosed only sporadically. SARS-CoV was, therefore, the most likely causal agent of SARS. The four SARS-CoV-infected macaques excreted SARS-CoV from nose, mouth, and pharynx from 2 days after infection. Three of four macaques developed diffuse alveolar damage, similar to that in SARS patients, and characterised by epithelial necrosis, serosanguineous exudate, formation of hyaline membranes, type 2 pneumocyte hyperplasia, and the presence of syncytia. SARS-CoV was detected in pneumonic areas by virus isolation and RT-PCR, and was localised to alveolar epithelial cells and syncytia by immunohistochemistry and transmission electron microscopy. INTERPRETATION: Replication in SARS-CoV-infected macaques of pneumonia similar to that in human beings with SARS, combined with the high prevalence of SARS-CoV infection in SARS patients, fulfill the criteria required to prove that SARS-CoV is the primary cause of SARS. PMID- 12892956 TI - Estimates of global and regional potential health gains from reducing multiple major risk factors. AB - BACKGROUND: Estimates of the disease burden due to multiple risk factors can show the potential gain from combined preventive measures. But few such investigations have been attempted, and none on a global scale. Our aim was to estimate the potential health benefits from removal of multiple major risk factors. METHODS: We assessed the burden of disease and injury attributable to the joint effects of 20 selected leading risk factors in 14 epidemiological subregions of the world. We estimated population attributable fractions, defined as the proportional reduction in disease or mortality that would occur if exposure to a risk factor were reduced to an alternative level, from data for risk factor prevalence and hazard size. For every disease, we estimated joint population attributable fractions, for multiple risk factors, by age and sex, from the direct contributions of individual risk factors. To obtain the direct hazards, we reviewed publications and re-analysed cohort data to account for that part of hazard that is mediated through other risks. RESULTS: Globally, an estimated 47% of premature deaths and 39% of total disease burden in 2000 resulted from the joint effects of the risk factors considered. These risks caused a substantial proportion of important diseases, including diarrhoea (92%-94%), lower respiratory infections (55-62%), lung cancer (72%), chronic obstructive pulmonary disease (60%), ischaemic heart disease (83-89%), and stroke (70-76%). Removal of these risks would have increased global healthy life expectancy by 9.3 years (17%) ranging from 4.4 years (6%) in the developed countries of the western Pacific to 16.1 years (43%) in parts of sub-Saharan Africa. INTERPRETATION: Removal of major risk factors would not only increase healthy life expectancy in every region, but also reduce some of the differences between regions. The potential for disease prevention and health gain from tackling major known risks simultaneously would be substantial. PMID- 12892957 TI - Herpes simplex encephalitis. PMID- 12892958 TI - Effect of viewing smoking in movies on adolescent smoking initiation: a cohort study. AB - BACKGROUND: Exposure to smoking in movies has been linked with adolescent smoking initiation in cross-sectional studies. We undertook a prospective study to ascertain whether exposure to smoking in movies predicts smoking initiation. METHOD: We assessed exposure to smoking shown in movies in 3547 adolescents, aged 10-14 years, who reported in a baseline survey that they had never tried smoking. Exposure to smoking in movies was estimated for individual respondents on the basis of the number of smoking occurrences viewed in unique samples of 50 movies, which were randomly selected from a larger sample pool of popular contemporary movies. We successfully re-contacted 2603 (73%) students 13-26 months later for a follow-up interview to determine whether they had initiated smoking. FINDINGS: Overall, 10% (n=259) of students initiated smoking during the follow-up period. In the highest quartile of exposure to movie smoking, 17% (107) of students had initiated smoking, compared with only 3% (22) in the lowest quartile. After controlling for baseline characteristics, adolescents in the highest quartile of exposure to movie smoking were 2.71 (95% CI 1.73-4.25) times more likely to initiate smoking compared with those in the lowest quartile. The effect of exposure to movie smoking was stronger in adolescents with non-smoking parents than in those whose parent smoked. In this cohort, 52.2% (30.0-67.3) of smoking initiation can be attributed to exposure to smoking in movies. INTERPRETATION: Our results provide strong evidence that viewing smoking in movies promotes smoking initiation among adolescents. PMID- 12892959 TI - Protection from natural infections with enterotoxigenic Escherichia coli: longitudinal study. AB - BACKGROUND: Enterotoxigenic Escherichia coli (ETEC) are an important cause of diarrhoea and diarrhoeal deaths in children living in developing countries and of travellers' diarrhoea. During the past 25 years, vaccine development efforts have been focused on induction of protective immunity against surface colonisation factors (CFs) and the heat-labile enterotoxin. Although vaccines that induce immunity to heat-labile toxin offer protection against diarrhoea from ETEC that produce this toxin, the benefit of including CF antigens remains uncertain. We aimed to estimate the protection that natural ETEC infections induce against new infections. METHODS: In Guinea-Bissau, we followed up 200 neonates until up to age 2 years, most of whom were breastfed throughout the study. We collected stool specimens from the children every week irrespective of whether they had diarrhoea. As a measure of protection, we used Cox regression models to estimate the change in infection rates after a primary ETEC infection. We thus estimated the protection attributable to CFs, toxins, and to any other factors that could be shared by ETEC with the same toxin-CF profile. FINDINGS: ETEC infections induced a 47% (95% CI 12 to 69) protection against new infections with ETEC that had the same toxin-CF profile; the corresponding estimate attributable to CFs was -1% (-40 to 27). Infection with heat-labile toxin-positive ETEC conferred a 45% ( 1 to 70) protection against symptomatic infections with ETEC positive for this toxin. INTERPRETATION: For breastfed children living in endemic areas, other antigens are substantially more important than CFs for induction of protective immunity against ETEC infection. PMID- 12892960 TI - Tinnitus after cycling. PMID- 12892961 TI - Treatment of SARS with human interferons. AB - Effective antiviral agents are needed to treat severe acute respiratory syndrome associated coronavirus (SARS-CoV) infection. We assessed the antiviral potential of recombinant interferons against two clinical isolates of SARS-CoV--FFM-1, from Frankfurt patients, and Hong Kong--replicated in Vero and Caco2 cells. Interferon beta was five to ten times more effective in Caco2 cells. Interferon alpha effectively inhibited SARS-CoV replication, but with a selectivity index 50-90 times lower than that for interferon beta. Interferon gamma was slightly better than interferon alpha in Vero cell cultures, but was completely ineffective in Caco2 cell cultures. Interferon beta could be useful alone or in combination with other antiviral drugs for the treatment of SARS. PMID- 12892962 TI - Neuropathological assessment of artemether-treated severe malaria. AB - In animals, high doses of intramuscular artemether and artemotil have been shown to cause an unusual pattern of selective damage to certain brainstem nuclei, especially those implicated in hearing and balance. We aimed to investigate whether a similar pattern arises in human adults. We examined the brainstems of adults who died after treatment with high dose artemether or quinine for severe falciparum malaria for evidence of a pattern of selective neuronal damage. Neuropathological findings were similar in recipients of quinine (n=15) and artemether (n=6; total artemether doses received 4-44 mg/kg). No evidence was recorded for artemether-induced neurotoxic effects. PMID- 12892963 TI - Gastric cancer. AB - The past decade has seen many advances in knowledge about gastric cancer. Notably, tumour biology and lymphatic spread are now better understood, and treatment by surgical and medical oncologists has become more standardised. Since refrigerators have replaced other methods of food conservation, Helicobacter pylori has become a factor in the cause of gastric cancer. Cancers that arise at the oesophagogastric junction might be further examples of wealth-associated disease. To tailor treatment better, the western hemisphere needs to borrow from the East by establishing screening programmes for early diagnosis, through careful surgical resection, and through detailed analysis of tumour spread. In Europe and the USA, most patients reach treatment with cancers already at an advanced stage. For these patients, three important randomised trials are underway that evaluate combined therapy. Cytostatic drugs, especially angiogenesis inhibitors have proved disappointing; however, basic research efforts to detect familial gastric cancers and to assess minimally residual disease look more hopeful. PMID- 12892964 TI - B-type natriuretic peptide in cardiovascular disease. AB - Natriuretic peptide hormones, a family of vasoactive peptides with many favourable physiological properties, have emerged as important candidates for development of diagnostic tools and therapeutic agents in cardiovascular disease. The rapid incorporation into clinical practice of bioassays to measure natriuretic peptide concentrations, and drugs that augment the biological actions of this system, show the potential for translational research to improve patient care. Here, we focus on the physiology of the natriuretic peptide system, measurement of circulating concentrations of B-type natriuretic peptide (BNP) and the N-terminal fragment of its prohormone (N-terminal BNP) to diagnose heart failure and left ventricular dysfunction, measurement of BNP and N-terminal BNP to assess prognosis in patients with cardiac abnormalities, and use of recombinant human BNP (nesiritide) and vasopeptidase inhibitors to treat heart failure. PMID- 12892965 TI - Knowledge into action for child survival. AB - The child survival revolution of the 1980s contributed to steady decreases in child mortality in some populations, but much remains to be done. More than 10 million children will die this year, almost all of whom are poor. Two-thirds of these deaths could have been prevented if effective child survival interventions had reached all children and mothers who needed them. Translation of current knowledge into effective action for child survival will require leadership, strong health systems, targeted human and financial resources, and modified health system to ensure that poor children and mothers benefit. A group of concerned scientists and policy-makers issues a call to action to leaders, governments, and citizens to translate knowledge into action for child survival. PMID- 12892966 TI - Can health programmes lead to mistreatment of sex workers? PMID- 12892967 TI - Can health programmes lead to mistreatment of sex workers? PMID- 12892968 TI - Mammography service screening and mortality. PMID- 12892969 TI - Acute shortage of nurses in India. PMID- 12892970 TI - Drug use during pregnancy: are risk classifications more dangerous than the drugs? PMID- 12892971 TI - New WHO chief promises greater commitment to HIV/AIDS. Lee names HIV/AIDS as his top priority and unveils the team that will help carry WHO's work forward. PMID- 12892972 TI - Improving health services: looking at the wrong link. PMID- 12892973 TI - Why the rise in Haemophilus influenzae type b infections? PMID- 12892974 TI - Why the rise in Haemophilus influenzae type b infections? PMID- 12892975 TI - UK nuclear-test veterans. PMID- 12892976 TI - ABO mismatching errors in heart transplants. PMID- 12892977 TI - Blood transfusions: a hidden source of lead exposure. PMID- 12892978 TI - Asia-Pacific and the millennium health targets. PMID- 12892979 TI - Brain natriuretic peptide in Chagas' disease: further insights. PMID- 12892980 TI - Otzi had a wound on his right hand. PMID- 12892981 TI - Europe refuses to match US cash for ailing Global Fund. PMID- 12892982 TI - Dannie Abse. PMID- 12892983 TI - Resistance to antiretrovirals is a growing concern. PMID- 12892984 TI - Feeding risk cut for HIV-infected women. PMID- 12892985 TI - Peter Lachmann. PMID- 12892986 TI - Money fever. PMID- 12892987 TI - The prevalence, impairment, impact, and burden of premenstrual dysphoric disorder (PMS/PMDD). AB - Currently it is estimated that 3-8% of women of reproductive age meet strict criteria for premenstrual dysphoric disorder (PMDD). Assessment of published reports demonstrate that the prevalence of clinically relevant dysphoric premenstrual disorder is probably higher. 13-18% of women of reproductive age may have premenstrual dysphoric symptoms severe enough to induce impairment and distress, though the number of symptoms may not meet the arbitrary count of 5 symptoms on the PMDD list. The impairment and lowered quality of life for PMDD is similar to that of dysthymic disorder and is not much lower than major depressive disorder. Nevertheless, PMS/PMDD is still under-recognized in large published epidemiological studies, as well as assessments of burden of disease. It is demonstrated here that the burden of PMS/PMDD as well as the disability adjusted life years (DALY) lost due to this repeated-cyclic disorder is in the same magnitude as major recognized disorders. Appropriate recognition of the disorder and its impact should lead to treatment of more women with PMS/PMDD. Efficacious treatments are available. They should reduce individual suffering and impact on family, society, and economy. PMID- 12892988 TI - Premenstrual syndrome and premenstrual dysphoric disorder: definitions and diagnosis. AB - Because of the prevalence, chronicity and distress caused by premenstrual symptoms (PMS), diagnosis and effective treatments are important information for clinicians. The DSM-IV requires at least five specified symptoms for premenstrual dysphoric disorder (PMDD), a severe dysphoric form of PMS, while the ICD-10 requires only one distressing symptom for a diagnosis of PMS. Many women who seek treatment fall between these two diagnostic approaches, and standard diagnostic criteria for clinically significant PMS are needed. A diagnosis of PMS consists of determining the timing of the symptoms in relation to menses, meaningful change between post- and premenstrual symptom severity and a clinically significant severity of the symptoms. A differential diagnosis to distinguish PMS from other medical and psychiatric conditions is important for appropriate treatment. No hormone or laboratory test indicates a PMS diagnosis. The current diagnostic standard requires confirmation of subjective symptom reports by prospective daily diaries. Diagnostic criteria for PMS must recognize the broad range of symptoms, the temporal pattern of the symptoms and the critical issue of symptom severity, which differentiates clinically significant PMS from normal menstrual cycle changes. PMID- 12892989 TI - A review of treatment of premenstrual syndrome and premenstrual dysphoric disorder. AB - Severe premenstrual syndrome (PMS) and, more recently, premenstrual dysphoric disorder (PMDD) have been studied extensively over the last 20 years. The defining criteria for diagnosis of the disorders according to the American College of Obstetricians and Gynecologists (ACOG) include at least one moderate to severe mood symptom and one physical symptom for the diagnosis of PMS and by DSM IV criteria a total of 5 symptoms with 1 severe mood symptom for the diagnosis of PMDD. There must be functional impairment attributed to the symptoms. The symptoms must be present for one to two weeks premenstrually with relief by day 4 of menses and should be documented prospectively for at least two cycles using a daily rating form. Nonpharmacologic management with some evidence for efficacy include cognitive behavioral relaxation therapy, aerobic exercise, as well as calcium, magnesium, vitamin B(6) L-tryptophan supplementation or a complex carbohydrate drink. Pharmacologic management with at least ten randomized controlled trials to support efficacy include selective serotonin reuptake inhibitors administered daily or premenstrually and serotonergic tricyclic antidepressants. Anxiolytics and potassium sparing diuretics have demonstrated mixed results in the literature. Hormonal therapy is geared towards producing anovulation. There is good clinical evidence for GnRH analogs with addback hormonal therapy, danocrine, and estradiol implants or patches with progestin to protect the endometrium. Oral contraceptive pills prevent ovulation and should be effective for the treatment of PMS/PMDD. However, limited evidence does not support efficacy for oral contraceptive agents containing progestins derived from 19-nortestosterone. The combination of the estrogen and progestin may produce symptoms similar to PMS, such as water retention and irritability. There is preliminary evidence that a new oral contraceptive pill containing low-dose estrogen and the progestin drospirenone, a spironolactone analog, instead of a 19 nortestosterone derivative can reduce symptoms of water retention and other side effects related to estrogen excess. The studies are in progress, however, preliminary evidence suggests that the drospirenone-containing pill called Yasmin may be effective the treatment of PMDD. PMID- 12892990 TI - The etiology, biology, and evolving pathology of premenstrual syndromes. AB - Menstrually related symptoms and disorders are multidimensional and affect diverse physiologic systems. Elucidation of the pathophysiologic mechanisms of these disorders should allow for a more precise diagnosis, and provide direction for targeted therapeutic interventions. Several biologic mechanisms that underlie menstrually related symptoms have been proposed. They focus mostly on gonadal hormones, their metabolites and interactions with neurotransmitters and neurohormonal systems, such as serotonin, GABA, cholecystokinin, and the renin angiotensin-aldosterone system. Altered responses of these systems to gonadal hormone's fluctuations during the menstrual cycle, as well as an increased sensitivity to changes in gonadal hormones may contribute to menstrually related symptoms in vulnerable women. Disrupted homeostasis and deficient adaptation may be core underlying mechanisms. Future directions for clinically-relevant progress include identification of specific subgroups of menstrually-related syndromes, assessment of the genetic vulnerability and changes in vulnerability along the life cycle, the diversified mechanisms by which vulnerability is translated into pathophysiology and symptoms, the normalization process as well as syndromes based and etiology-based clinical trials. PMID- 12892992 TI - Basic aspects of the biochemical reactivity of 4-hydroxynonenal. AB - 4-hydroxynonenal (HNE), a major lipid peroxidation product of n-6 polyunsaturated fatty acids, which was discovered by the late Hermann Esterbauer, is a remarkable trifunctional molecule. Both the hydroxy group and the conjugated system consisting of a C=C double bond and a carbonyl group contribute to the high reactivity of HNE. Most of the biochemical effects of HNE can be explained by its rapid reactions with thiol and amino groups. Among the primary reactants for HNE are the amino acids cysteine, histidine and lysine, which--either free or protein bound--undergo readily Michael additions to the C=C bond. After this primary reaction, which confers rotational freedom to the C2-C3 bond, secondary reactions may occur involving the carbonyl and the hydroxy group. Primary amines may alternatively react with the carbonyl group to form Schiff bases. Reactions which do not fit into this scheme are the oxidation and the reduction respective of the carbonyl group and the epoxidation of the C=C double bond. Examples will be presented for the interaction of HNE with various classes of biomolecules such as proteins and peptides, lipids and nucleic acids and the biochemical consequences will be discussed. PMID- 12892993 TI - Genotoxicity of HNE. AB - Since previous investigations on the genotoxicity of 4-hydroxynonenal (HNE) were carried out with prokaryotic systems or eukaryotic cell lines which may not adequately reflect the response of cells in vivo due to differences in the metabolism, the genotoxic potential of HNE was further evaluated in primary cells (hepatocytes) and cell clones of cerebral endothelial cells expressing specific functions, i.e. blood-brain barrier (BBB) and capillary formation associated phenotypes. Treatment of hepatocytes with HNE induced statistically significant levels of SCE at concentrations >/=0.1 microM, micronuclei at concentrations >/=1 microM and chromosomal aberrations at a concentration of 10 microM. Treatment of cloned cerebral microvascular endothelial cells induced significantly elevated levels of chromosomal aberrations at concentrations >/=1 microM and micronuclei at concentrations >/=10 microM in both cEC phenotypes, compared to the controls. Additionally, cytotoxicity was observed at a concentration of 50 microM HNE and was significantly higher in type II cells. These results indicate that cells expressing differentiated functions representative for the in vivo situation react more sensitive to HNE than cell lines, and may reflect the sensitivity of the target cells. The different response with respect to the endpoints of genotoxicity tested most probably depends on the different metabolizing capacities and thus the action of different metabolites of HNE. PMID- 12892994 TI - Intracellular metabolism of 4-hydroxynonenal. AB - 4-hydroxynonenal (HNE) is a major aldehydic product of lipid peroxidation known to exert a multitude of biological, cytotoxic, and signal effects. Mammalian cells possess highly active pathways of HNE metabolism. The metabolic fate of HNE was investigated in various mammalian cells and organs such as hepatocytes, intestinal enterocytes, renal tubular cells, aortic and brain endothelial cells, synovial fibroblasts, neutrophils, thymocytes, heart, and tumor cells. The experiments were carried out at 37 degrees C at initial HNE concentrations between 1 microM--that means in the range of physiological and pathophysiologically relevant HNE levels--to 100 microM. In all cell types which were investigated, 90-95% of 100 microM HNE were degraded within 3 min of incubation. At 1 microM HNE the physiological blood serum level of about 0.1-0.2 microM was restored already after 10-30 s. As primary products of HNE in hepatocytes and other cell types the glutathione-HNE-1:1-conjugate, the hydroxynonenoic acid and the corresponding alcohol of HNE, the 1,4 dihydroxynonene, were identified. Furthermore, the beta-oxidation of hydroxynonenoic acid including the formation of water was demonstrated. The quantitative share of HNE binding to proteins was low with about 2-8% of total HNE consumption. The glycine-cysteine-HNE, cysteine-HNE adducts and the mercapturic acid from glutathione-HNE adduct were not formed in the most cell types, but in kidney cells and neutrophils. The rapid metabolism underlines the role of HNE degrading pathways in mammalian cells as important part of the secondary antioxidative defense mechanisms in order to protect proteins from modification by aldehydic lipid peroxidation products. PMID- 12892995 TI - Fate of 4-hydroxynonenal in vivo: disposition and metabolic pathways. AB - Due to the cytotoxicity of 4-hydroxynonenal (HNE), and to the fact that this major product of lipid peroxidation is a rather long-living compound compared with reactive oxygen species, the capability of organisms to inactivate and eliminate HNE has received increasing attention during the last decade. Several recent in vivo studies have addressed the issue of the diffusion, kinetics, biotransformation and excretion of HNE. Part of these studies are primarily concerned with the toxicological significance of HNE biotransformation and more precisely with the metabolic pathways by which HNE is inactivated and eliminated. The other aim of in vivo metabolic study is the characterisation of end metabolites, especially in urine, in order to develop specific and non-invasive biomarkers of lipid peroxidation. When HNE is administered intravenously or intraperitoneally, it is mainly excreted into urine and bile as conjugated metabolites, in a proportion that is dependent on the administration route. However, biliary metabolites undergo an enterohepatic cycle that limits the final excretion of faecal metabolites. Only a very low amount of metabolites is found to be bound to macromolecules. The main urinary metabolites are represented by two groups of compounds. One comes from the mercapturic acid formation from (i) 1,4 dihydroxynonene-glutathione (DHN-GSH) which originates from the conjugation of HNE with GSH by glutathione-S-transferases and the subsequent reduction of the aldehyde by a member of aldo-keto reductase superfamily; (ii) the lactone of 4 hydroxynonanoic-GSH (HNA-lactone-GSH) which originates from the conjugation of HNE followed by the oxidation of the aldehyde by aldehyde dehydrogenase; (iii) HNA-GSH which originates from the hydrolysis of the corresponding lactone. The other one is a group of metabolites issuing from the omega-hydroxylation of HNA or HNA-lactone by cytochromes P450 4A, followed eventually, in the case of omega oxidized-HNA-lactone, by conjugation with GSH and subsequent mercapturic acid formation. Biliary metabolites are GSH or mercapturic acid conjugates of DHN, HNE and HNA. Stereochemical aspects of HNE metabolism are also discussed. PMID- 12892996 TI - HNE--signaling pathways leading to its elimination. AB - The oxidation of polyunsaturated fatty acids results in the production of HNE, which can react through both non-enzymatic and enzyme catalyzed reactions to modify a number of cellular components, including proteins and DNA. Multiple pathways for its enzyme catalyzed elimination include oxidation of the aldehyde to a carboxylic acid, reduction of the aldehyde to an alcohol, and conjugation of the carbon-carbon double bond to glutathione (GSH). Interestingly, the enzymes that result in HNE elimination are induced by HNE itself although the chemical mechanism for signaling is not well understood. One of the striking effects of HNE is that after a transient decrease in GSH, synthesis of GSH is elevated through induction of glutamate cysteine ligase (GCL), which catalyzes the first step in de novo synthesis of GSH. GCL has two subunits, which are transcriptionally regulated by a wide variety of agents, including oxidants and electrophiles, such as HNE, which elevates both. The transcriptional regulation of GCL has been the subject of many investigations yielding a complex picture in which the pathways for up-regulation of the subunits appear to be independent and vary with inducing agent and cell type. We have found that in human bronchial epithelial cells, HNE acts through AP-1 activation with signaling through the JNK pathway, and that neither the ERK nor p38(MAPK) pathways is involved. With these results we review what is currently known about the signaling mechanisms for removal of HNE, focusing principally on conjugation mechanisms involving GSH. PMID- 12892997 TI - The proteasomal system and HNE-modified proteins. AB - Metabolic processes and environmental conditions cause the constant formation of oxidizing species over the lifetime of cells and organisms. This leads to a continuous oxidation of intracellular components, including lipids, DNA and proteins. During the extensively studied process of lipid peroxidation, several reactive low-molecular weight products are formed, including reactive aldehydes as 4-hydroxynonenal (HNE). These aldehydic lipid peroxidation products in turn are able to modify proteins. The degradation of oxidized and oxidatively modified proteins is an essential part of the oxidant defenses of cells. The major proteolytic system responsible for the removal of oxidized cytosolic and nuclear proteins is the proteasomal system. The proteasomal system by itself is a multicomponent system responsible for the degradation of the majority of intracellular proteins. It has been shown that some, mildly cross-linked, HNE modified proteins are preferentially degraded by the proteasome, but extensive modification with this cross-linking aldehyde leads to the formation of protein aggregates, that can actually inhibit the proteasome. This review summarizes our knowledge of the interactions between lipid peroxidation products, proteins, and the proteasomal system. PMID- 12892998 TI - Role of PKC-dependent pathways in HNE-induced cell protein transport and secretion. AB - The beta isoforms of protein Kinase C (PKC) are closely involved in the regulation of cell protein transport and secretion. We have shown in different cellular types that treatment with HNE in a concentration range detectable in many pathophysiological conditions is able to induce selective activation of betaPKCs through direct interaction between the aldehyde and these isoenzymes. In isolated rat hepatocytes this specific isoenzyme activation plays a key role in the transport of procathepsin D from the trans-Golgi network to the endosomal lysosomal compartment and in the exocytosis of mature cathepsin D. In NT2 neurons, HNE-mediated betaPKC activation induces an increase in intracellular amyloid beta production, without affecting full-length amyloid precursor protein expression. In a mouse macrophage-like cell line, the same beta isoform activation increases the release of the MCP-1 chemokine. Thus, pathophysiological HNE concentrations (0.1-1 microM) derived from a slight imbalance of the redox state are able to alter protein trafficking through beta PKC activation. These results suggest that mild oxidative stress and the PKC signal transduction pathway are closely involved in the pathophysiology of many diseases caused by changes in protein trafficking and release. PMID- 12892999 TI - 4-hydroxy-2-nonenal as a COX-2 inducer. AB - 4-hydroxy-2-nonenal (HNE) activates a variety of signaling pathways. We have recently evaluated the effect of oxidized fatty acid metabolites on cyclooxygenase-2 (COX-2) induction in rat liver epithelial RL34 cells and found that, among the compounds tested, HNE most dramatically induced COX-2. A p38 mitogen-activated protein kinase (p38 MAPK) pathway has been shown to play a key role in the mechanism of the HNE-induced COX-2 expression. It appears that the HNE-induced activation of p38 MAPK leads to the stabilization of COX-2 mRNA. PMID- 12893000 TI - Role of 4-hydroxynonenal in stress-mediated apoptosis signaling. AB - In this mini review we summarize recent studies from our laboratory, which show the involvement of 4-hydroxynonenal (4-HNE) in cell cycle signaling. We demonstrate 4-HNE induced apoptosis in various cell lines is accompanied with c Jun-N-terminal kinase and caspase-3 activation. Cells exposed to mild, transient, heat or oxidative stress acquire capacity to exclude intracellular 4-HNE at a faster rate by inducing hGST5.8 which conjugate 4-HNE to GSH, and RLIP76 which mediates the ATP-dependent transport of the GSH-conjugate of 4-HNE. The cells preconditioned with mild transient stress acquire resistance to H(2)O(2) and 4 HNE induced apoptosis by excluding intracellular 4-HNE at an accelerated pace. Furthermore, a decrease in intracellular concentration of 4-HNE achieved by transfecting cells with mGSTA4-4 or hGSTA4-4 results in a faster growth rate. These studies strongly suggest a role of 4-HNE in stress mediated signaling. PMID- 12893001 TI - 4-hydroxynonenal triggers multistep signal transduction cascades for suppression of cellular functions. AB - 4-hydroxynonenal (HNE), an aldehyde product of membrane lipid peroxidation, has been suggested to mediate a number of oxidative stress-linked pathological events in humans, including cellular growth inhibition and apoptosis induction. Because HNE is potentially reactive to a number of both cell surface and intracellular proteins bearing sulfhydryl, amino and imidazole groups, it seems that there are multiple signal transduction cascades. Here we briefly review the HNE-triggered signal transduction cascades that lead to suppression of cellular functions and to cell death, based mainly on our own recent study results. We first showed that formation of HNE-cell surface protein adducts, which mimicked ligand-cell surface receptor binding, induced activation of receptor-type protein tyrosine kinases such as epithelial growth factor receptor (EGFR) and that this caused growth inhibition through a cascade of activation of EGFR, Shc and ERK. Next, we showed that HNE-mediated scavenging of cellular glutathione led to activation of caspases and to DNA fragmentation through a Fas-independent and mitochondria linked pro-apoptotic signal pathway. More recently, we have obtained evidence that the HNE-triggered signal cascade for caspase activation encounters complex positive feedback regulatory mechanisms that are linked to the inhibition of anti apoptotic signals and are dependent on caspase activity. Underlying multiple regulatory mechanisms, including mechanisms of activation of Akt dephosphorylating PP2A activity, activities of protein tyrosine kinases have been shown to be biphasically controlled by HNE. In addition, we have obtained results suggesting that HNE inhibits phosphorylation of IkappaB, possibly by targeting some elements upstream of IkappaB, which might downregulate the NF-kappaB mediated cellular responses, including serum deprivation-induced iNOS expression and generation of anti-apoptotic signals. These results suggest that HNE reacts with multiple cell surface and intracellular sites for triggering a network of signal transduction that is ultimately focused on suppression of cellular functions. PMID- 12893002 TI - Cholesteryl ester oxidation products in atherosclerosis. AB - Lipid oxidation products are formed at sites of increased oxidant stress and have been shown to accumulate in atherosclerotic lesions. Although recent studies have focused on the formation and metabolism of oxidized lipids, very little is known about their biological activities and possible (patho)physiological functions. Oxidation of cholesteryl esters containing unsaturated fatty acids leads to the formation of hydroperoxides that are either reduced to alcohols or degrade into biologically active "core-aldehydes". In this review, the mechanisms of formation and metabolic fate of oxidized cholesteryl esters, their occurrence, as well as possible biological activities are discussed. Based on the current knowledge, cholesteryl ester oxidation leads to the formation of biologically active substances, which could actively contribute to the progression of atherosclerotic lesions and their resulting complications. PMID- 12893003 TI - Oxidized LDL and 4-hydroxynonenal modulate tyrosine kinase receptor activity. AB - Among the diverse risk factors involved in atherosclerosis, LDL are thought to become atherogenic after undergoing oxidative modifications, characterized by oxidized lipid formation and structural alterations of apoB. Oxidized LDL alter various signaling pathways and exhibit a broad range of biological responses including inflammation, gene expression, cell proliferation or apoptosis. The biological effects of oxidized LDL are related to the presence of peroxidation products such as hydroperoxides, lysophosphatidylcholines, oxysterols and aldehydes.4-Hydroxynonenal (HNE) is one of the most abundant aldehydes formed during the oxidation of polyunsaturated fatty acids in LDL and in membranes. It is able to react with thiols and free amino group residues of proteins. HNE is involved in apoB modifications that alter LDL metabolism and cell protein-adduct formation which may mediate in part the biological effects of oxidized LDL. We report here that HNE delivered to cells by oxidized LDL reacts with cellular proteins, for instance with tyrosine kinase receptors (RTK) such as EGFR and PDGFR. HNE induces in vitro derivatization and tyrosine phosphorylation of RTK (the fine molecular mechanism and conformational changes remain to be elucidated). In intact living cells, oxidized LDL (and pure HNE) trigger HNE adduct formation and activation of PDGFR and EGFR, through an antioxidant insensitive and reactive oxygen species independent mechanism. The presence of HNE-PDGFR adducts in atherosclerotic areas lead one to hypothesize that oxidized lipids may also react in vivo with membrane RTK, thereby disturbing their cellular functions. PMID- 12893004 TI - 4-hydroxynonenal from pathology to physiology. AB - 4-hydroxynonenal is a major product of lipid peroxidation. It was firstly studied under the point of view of its toxicity, as it is an easily diffusable substance, thought to be able to explain the "far damages" seen in conditions of increased lipid peroxidation. Really, when used at concentration from 10 microM to 1 mM, usually referred to as high concentrations, the aldehyde is able to produce strong inhibitions of several enzymatic activities. When used, however, at concentration of 1 microM or lower, it displays a lot of activities regarding especially cell multiplication and differentiation. As the concentrations indicated above are usually found in normal tissues, these effects may be considered as physiological. As a low level of lipid peroxidation exists in normal tissues, the aldehyde displays signalling activities in normal cells. Among them, it is to consider the stimulation of neutrophil chemotaxis, the strong activation of plasmamembrane adenylate kinase, the strong activation of membrane phospholipase C, both in hepatocytes and neutrophils, the block in the expression of the oncogene c-myc in human leukemic cells, accompanied by differentiation of the same cells, the effects on the cyclins and the activity of E2F transcription factor, the strong increase of the expression of the gene for procollagen alfa1(I), occurring due to the activation of the c-jun/junkinases/AP 1 pathway. Moreover, it is able to block the activity of the PDGF-beta receptor. The last facts allow to think that a hydroxynonenal pathway works in the production of fibrosis. PMID- 12893005 TI - 4-hydroxynonenal and transforming growth factor-beta1 expression in colon cancer. AB - In vivo studies on human colon adenocarcinoma showed decreased transforming growth factor-beta1 (TGF-beta1) antiproliferative cytokine content in tumour tissue related to malignancy progression, with a corresponding decrease in lipid peroxidation aldehydic end-product, 4-hydroxynonenal (HNE). The tumour mechanism to escape TGF-beta1-mediated growth inhibition may be due to an altered TGF-beta1 receptor system. Subsequent in vitro analyses showed a differential distribution of TGF-beta1 receptors depending on the human colon cancer cell line considered (CaCo-2 or HT-29): compared to HT-29 cells, CaCo-2 cells showed a decrease of the two main TGF-beta1 receptors, RI and RII. Notwithstanding their partial TGF-beta1 RI and RII deficiency, treatment of CaCo-2 cells with adequate doses of the cytokine (10 ng/ml) was able to induce apoptosis. Of note, co-treatment of these cells with 1 microM HNE increased the apoptotic effect. The constant low concentration of TGF-beta1 in the tumour mass may be related to the low content of antiproliferative HNE observed in colon cancer: the latter phenomenon, which reduces TGF-beta1 production in the tumour area, may represent a favourable condition for neoplastic progression. The enhancement of TGF-beta1-induced apoptosis by HNE in CaCo-2 cells supports this hypothesis. The different transcriptional components regulated by the distinct signaling pathways of these two molecules might be proposed; in particular, crosstalk between the MAPK and the Smad pathway could modulate and co-operate in the transcription of target genes involved in regulation of cell proliferation. PMID- 12893006 TI - 4-hydroxynonenal as a bioactive marker of pathophysiological processes. AB - The review is focused on the currently major aspect of 4-hydroxynonenal (HNE) research--studies that combine biological activities of the aldehyde together with the methods of its identification in cells and tissues. Because there were some excellent reviews on HNE published in recent years, starting in 1990 and 1991 with supreme reviews done by Hermann Esterbauer, who discovered the aldehyde, and colleagues from the Institute of Biochemistry in Graz, this article pays most of attention to the most recent articles, published in the last 15 months. Additionally, an overview on the relevance of HNE is given with respect to the research and publication trends in the period of 10 years (1993-2002) according to the data in the Current Contents and Medline data bases. It is obvious that HNE started in 1993 as a "toxic product of lipid peroxidation" and "second toxic messenger of free radicals", to become in 2002 a reliable marker of oxidative stress, a possible causative agent of several diseases (such as Alzheimer's disease), growth modulating factor and a signaling molecule. Novel analytical methods developed suitable pathways for HNE to become a clinically applicable marker of lipid peroxidation on one side and on the other a standardized parameter of food quality control. As it is also present physiologically in various cells and tissues, it is likely that HNE will soon become one of the most attractive factors for those who search for a small and reactive molecular link between genomics and proteomics. PMID- 12893007 TI - 4-hydroxynonenal and neurodegenerative diseases. AB - The development of oxidative stress, in which production of highly reactive oxygen species (ROS) overwhelms antioxidant defenses, is a feature of many neurological diseases: ischemic, inflammatory, metabolic and degenerative. Oxidative stress is increasingly implicated in a number of neurodegenerative disorders characterized by abnormal filament accumulation or deposition of abnormal forms of specific proteins in affected neurons, like Alzheimer's disease (AD), Pick's disease, Lewy bodies related diseases, amyotrophic lateral sclerosis (ALS), and Huntington disease. Causes of neuronal death in neurodegenerative diseases are multifactorial. In some familiar cases of ALS mutation in the gene for Cu/Zn superoxide dismutase (SOD1) can be identified. In other neurodegenerative diseases ROS have some, usually not clear, role in early pathogenesis or implications on neuronal death in advanced stages of illness. The effects of oxidative stress on "post-mitotic cells", such as neurons may be cumulative, hence, it is often unclear whether oxidative damage is a cause or consequence of neurodegeneration. Peroxidation of cellular membrane lipids, or circulating lipoprotein molecules generates highly reactive aldehydes among which one of most important is 4-hydroxynonenal (HNE). The presence of HNE is increased in brain tissue and cerebrospinal fluid of AD patients, and in spinal cord of ALS patients. Immunohistochemical studies show presence of HNE in neurofibrilary tangles and in senile plaques in AD, in the cytoplasm of the residual motor neurons in sporadic ALS, in Lewy bodies in neocortical and brain stem neurons in Parkinson's disease (PD) and in diffuse Lewy bodies disease (DLBD). Thus, increased levels of HNE in neurodegenerative disorders and immunohistochemical distribution of HNE in brain tissue indicate pathophysiological role of oxidative stress in these diseases, and especially HNE in formation of abnormal filament deposites. PMID- 12893008 TI - Hydroxynonenal, toxic carbonyls, and Alzheimer disease. AB - Cytoskeletal disruption is one of the distinguishing characteristics of the vulnerable neurons in Alzheimer disease (AD). It has been suggested that these cytoskeletal changes occur secondarily to covalent modifications of the protein components. Despite the abundance and probable importance of these changes, there has been very little data regarding the identity of the modified proteins or the precise chemistry of the modifications. Here we review a specific type of modification, namely carbonylation of proteins, which has been shown to be a common result of cellular oxidative stress. Hopefully, the following discussion will help elucidate the relationship between oxidative stress, protein modification and the pathogenesis of AD. PMID- 12893010 TI - Resolving public conflict in site selection process-a risk communication approach. AB - In Japan, conflicts regarding the siting of waste disposal facilities occur frequently. In particular, siting of incinerators and landfills has become increasingly difficult because the public is highly concerned about the dioxin issues. Inefficient siting of waste disposal facilities causes several social problems, such as the shortage of waste treatment and disposal facilities, the rising of waste management costs and an increase in the consumption of resources. While dealing with a similar situation, the Chemical Society of Japan adopted a risk communication technique successfully. Hence, the pragmatic use of a risk communication technique is proposed to avoid conflicts and for a smooth information exchange to seek cooperation in waste management. In order to achieve this, a study was conducted to resolve conflicts between residents and the municipality for the selection of site for a solid waste treatment and disposal facility. This study aims to discuss the subject of risk communication for the waste disposal system in Japan. This study is performed through personal interviews and a questionnaire covering opposing parties in the town. As a result of the survey, a risk communication approach for a waste treatment and disposal system is presented in the paper addressing issues such as building of social trust, pragmatic use of the communication process, installation of credible information sources, and environmental education and awareness. PMID- 12893011 TI - Environmental management by the learning curve. AB - This is a futuristic appreciation of waste management challenges and their solution by means of good management models. A literature review, administrative initiatives, research results, and experiences from practice are combined in this study to render an evolutionary picture of the change in paradigm relative to municipal solid waste possible to occur between 2000 and 2025. The principal stages of progress in the 25 years studied were: the correct characterization of municipal solid waste as a function of geographical location and recycling potential, the divided collection model and its corresponding learning curve, correct opportunity cost accounting tools, and the generally admitted and accepted changeover of municipal solid waste treatment from a technical to a management problem. It is reported that as a result of this progress, the municipal landfill is a species in extinction. Regional landfills with long life spans are the rule in 2025. PMID- 12893012 TI - Assessment of the environmental impact of management measures for the biodegradable fraction of municipal solid waste in Sao Paulo City. AB - There is increasing concern about landfilling of biodegradable wastes. Therefore, biological treatment processes such as composting and biogasification have been considered as alternative strategies for managing those wastes. In this work, life cycle assessment was employed to compare the environmental impacts of landfilling, composting, and biological treatment of municipal solid waste in Sao Paulo City, Brazil. Energy consumption, recovered resources, and emissions to air and water were quantified and analyzed in terms of their potential contribution to global warming, acidification, and nutrient enrichment impact. The results demonstrated that processes that require high levels of energy consumption, such as wastewater treatment, play an important role in the outcome of environmental impact potentials. It was found that the landfilling of all waste is generally the worst strategy from an environmental point of view. However, significant reductions in the resulting impacts can be accomplished through biogasification and composting of the biodegradable fraction. Regarding composting, the application of a biofilter for gas treatment reduced significantly the gaseous emissions. PMID- 12893013 TI - Thickness of the buffer layer in the 'reaction front' model of solid-state digestion. AB - Conventional models of anaerobic digestion assume dispersed reaction, in the aqueous phase and at solid-liquid interfaces. However, a second, localized mechanism might operate in a bed of nutrient-rich wastes, such as occurs in many landfills and some waste digesters. A 'reaction front' (comprising two distinct reaction zones separated by a buffer zone) might develop around 'seed bodies' (local concentrations of methanogenic inoculum) then propagate through the waste. The initial conditions, notably the mode of seeding, would determine whether which mechanism is dominant. This paper presents a method of predicting the thickness of such a front and indicates a thickness of the order of 3.5 cm. This would suggest that the seed material must be present in the form of 'seed bodies' of over 7 cm diameter or layers over 7 cm thick. The probability of the formation of a reaction front is discussed, as are the implications for operational practice. PMID- 12893014 TI - The effect of lignin and sugars to the aerobic decomposition of solid wastes. AB - A series of experimental runs were conducted from 1995 to 1999 in Madison (WI, USA) with the goal to investigate the biodegradation process of seven (7) solid waste components and mixtures of them under near optimal aerobic conditions. It was shown that substrates with high initial lignin contents or high initial HWSM contents were observed to have relatively low and high degradation extents, respectively. Two linear equations were derived that correlate degradation extent (as indicated by the volatile solids reduction) to initial lignin and initial HWSM contents separately. The lignin equation was compared to a similar equation previously developed for anaerobic environments by Chandler et al. (Predicting methane fermentation biodegradability. In: Biotechnology and Bioengineering Symposium No. 10 (1980) New York: John Wiley & Sons). With comparison to the Chandler formula, lignin was found to be less inhibitory to the overall substrate decomposition in aerobic environments compared to anaerobic ones. Cellulose loss contributed to a higher than 50% to the overall dry mass loss for all substrates studied. In addition, the cellulose to lignin (C/L) ratio appeared to be a relatively accurate compost maturity indicator, since it reduced to a value less than 0.5 for most substrates that had reached their degradation extent. PMID- 12893015 TI - Solid waste management practices and review of recovery and recycling operations in Turkey. AB - This paper provides a general overview of solid waste data and management practices employed in Turkey during the last decade. Municipal solid waste statistics and management practices including waste recovery and recycling initiatives have been evaluated. Detailed data on solid waste management practices including collection, recovery and disposal, together with the results of cost analyses, have been presented. Based on these evaluations basic cost estimations on collection and sorting of recyclable solid waste in Turkey have been provided. The results indicate that the household solid waste generation in Turkey, per capita, is around 0.6 kg/year, whereas municipal solid waste generation is close to 1 kg/year. The major constituents of municipal solid waste are organic in nature and approximately 1/4 of municipal solid waste is recyclable. Separate collection programmes for recyclable household waste by more than 60 municipalities, continuing in excess of 3 years, demonstrate solid evidence for public acceptance and continuing support from the citizens. Opinion polls indicate that more than 80% of the population in the project regions is ready and willing to participate in separate collection programmes. The analysis of output data of the Material Recovery Facilities shows that, although paper, including cardboard, is the main constituent, the composition of recyclable waste varies strongly by the source or the type of collection point. PMID- 12893016 TI - Combustion studies of high moisture content waste in a fluidised bed. AB - The combustion of three high moisture content waste materials in a fluidised bed combustor has been investigated and a comparison with co-firing of these materials with coal in the same combustor has been made. Waste materials burnt were olive oil waste, municipal solid waste and potato, which is representative of vegetable waste. Mixtures of up to 20% mass concentration water in the waste were fed to the combustor. Above that value the moisture content was too high to sustain combustion without addition of coal. Measurements of CO, NOx, SO2 temperatures were made and the carbon combustion efficiency evaluated. Co-firing with coal resulted in markedly higher combustion efficiencies with an increase of approximately 10-80% when burning the simulated MSW. However, this was much lower than the value of 93% when coal was burnt on its own. It was also much lower than the value obtained, average 90%, when co-firing potato and olive oil waste with coal and there was little difference in the combustion efficiency between the two types of waste and with increasing moisture content. It was concluded that the high ash content of the simulated MSW 26%, compared with 5% in the other two waste materials resulted in slower burning and consequently the char particles were elutriated from the bed without being fully burnt. In term of gaseous emissions during co-combustion, CO emission is relatively insensitive to change in waste fraction. While emission of SO2 can be reduced as the waste fraction increases as a result of fuel-S dilution. But in terms of percent fuel-S converted, it is actually increased by increasing waste fraction. Emissions of NO and N2O increase slightly with MSW fraction. PMID- 12893017 TI - Advanced physico-chemical treatment experiences on young municipal landfill leachates. AB - In this study, Membrane Filtration (UF+RO), Struvite (MAP) precipitation and ammonia stripping alternatives were studied on biologically pre-treated Landfill Leachate. The results indicated that the system including the Upflow Anaerobic Sludge Blanket Reactor (UASBR) and Membrane Reactors (UF+RO) has been offered as an appropriate treatment alternative for young landfill leachates. This system provided high removals of COD, colour and conductivity (>98-99%). For ammonia removal, struvite precipitation was applied at the stoichiometric ratio (Mg:NH4:PO4=1:1:1) to anaerobically pre-treated raw landfill leachate effluent having an influent ammonium concentration of 2240 mg/l. Maximum ammonium nitrogen removal was observed as 85% at pH of 9.2. In ammonia stripping following 2 h of aeration, the removal was 72% at pH=12 while the removals were around 20% at pH=10 and pH=11. When membrane reactor, and struvite precipitation or ammonia stripping was applied to anaerobically pre-treated effluents, the results indicated that each system could be used as an appropriate post-treatment option for young landfill leachates. In economic aspect, ammonia stripping was found as the cheapest alternative with high ammonium removal. However, when both high COD and ammonium removals were to be achieved membrane technology such as UF+RO (SW) could be considered as the most appropriate system due to the fact that COD removal could be obtained very low by ammonia stripping. PMID- 12893018 TI - Low-cost treatment of landfill leachate using peat. AB - The EU Landfill Directive obliges member states to collect and treat leachate from landfill sites. In regions of high population density, this is commonly achieved through discharge of the leachate to the municipal sewerage system. In Ireland, rural landfills can be a long distance from a suitable sewerage system, resulting in high transportation costs. On-site treatment systems, when used elsewhere, are mainly aerobic treatment systems, which are costly to construct and operate. There is a particular need for low-cost, low-maintenance leachate treatment systems for small low-income landfills, and for closed landfills, where long-term running costs of aerobic systems may be unsustainable. In 1989, this research work was initiated to investigate the use of local peat for the treatment of leachate from a small rural landfill site. In 1997, following the award of grant-aid under the EU LIFE Programme, a full-scale leachate treatment plant was constructed, using local un-drained peat as the treatment medium. When the LIFE Project ended in February 2001, leachate treatment research continued at the site using a pre-treated peat as the treatment medium. The treatment levels achieved using both types of peat are discussed in this paper. It is concluded that landfill leachate may be successfully treated using a low-cost peat bed to achieve almost 100% removal of both BOD and ammonia. PMID- 12893019 TI - How vital is the "lack of funding" in effective environmental management in Turkey? AB - The conflict experienced in Turkey between adequate legal and legislative arrangements with weak capabilities of policy implementation and enforcement is a typical scenario experienced in developing countries. The difficulties faced in environmental management is increased by funding issues. Although environmental management is directly influenced by problems associated with the distribution of public funds, lack of funding is not the sole reason responsible for the mismanagement of resources. The paper seeks to answer whether the lack of funding allocated to ameliorate environmental degradation and pollution is the main reason for an environmental crisis situation in Turkey. Turkey needs to ensure implementation of the basic obligations stated in its environmental policy with respect to national and international issues. Specific obstacles Turkey needs to overcome for more effective environmental management will be examined within this context. PMID- 12893020 TI - Management of heparin-induced thrombocytopenia: a critical comparison of lepirudin and argatroban. AB - Heparin-induced thrombocytopenia (HIT) is a transient hypercoagulability state initiated, paradoxically, by the anticoagulant, heparin. It is characterized by antibody-induced activation of platelets, leading to thrombin generation. Many patients with HIT develop thrombosis; even when heparin is stopped because of "isolated HIT" detected during routine platelet count monitoring, 25-50% of patients subsequently develop symptomatic thrombosis. Thus, an alternative anticoagulant should be substituted for heparin when HIT is strongly suspected. Two direct thrombin inhibitors (DTIs), lepirudin and argatroban, have been studied for prevention and treatment of thrombosis in HIT patients. Lepirudin is a polypeptide that binds irreversibly to the fibrin-binding and catalytic sites on thrombin (bivalent inhibitor). In contrast, argatroban is a synthetic, small molecule DTI that binds reversibly to the catalytic site alone (univalent inhibitor). Results of historically controlled clinical trials suggest both agents are effective for preventing and treating thrombosis in HIT. However, these agents have not been compared directly, and important differences in study design limit conclusions from indirect comparison. For example, lepirudin was given for 12-14 days (mean) in treatment studies of thrombosis complicating HIT, whereas argatroban was given only for 6-7 days, a difference that could explain apparent lower thrombosis rates (and greater bleeding) with lepirudin. Recently, the transition from DTI therapy to oral anticoagulation in patients with deep venous thrombosis (DVT) complicating HIT has been identified as a risk period for coumarin-induced venous limb gangrene. Thus, the DTI should be given alone during acute HIT, with oral anticoagulants deferred until substantial resolution of the thrombocytopenia has occurred. PMID- 12893021 TI - Myeloproliferative syndromes and the associated risk of coronary artery disease. AB - INTRODUCTION: Of the major myeloproliferative syndromes (MPS) [polycythemia vera (PV), essential thrombocythemia (ET), chronic myeloid leukemia (CML) and myelofibrosis (MF)], PV and ET are reported to be associated with increased thrombotic complications. However, the relationship between these myeloproliferative syndromes and coronary artery disease (CAD) is unclear. METHODS: We performed a retrospective chart review to evaluate the prevalence of CAD in patients with diagnosed with MPS between 1991 and 2001. RESULTS: One hundred and eighty-one patients (100 males, 81 females) with a mean age of 72.5 years were included. Twenty-nine patients, 19 males and 10 females (16%, 95% CI: 12.0-24.0) had CAD. These included 6/53 (11.3%, 95% CI: 1.5-20.2) patients with CML, 1/26 (3.8%, 95% CI: -4.4 to 12.8) patients with PV, 5/30 (16.7%, 95% CI: 2.5 30.8) patients with ET, 3/7 (42.9%, 95% CI: 12.3-87.7) patients with MF and 14/65 (21.5%, 95% CI: 13.1-37.8) patients with co-existent MPS. Comparing the risk of CAD with CML as a baseline, MF had an OR of 8.2 (p < 0.01, 95% CI: 1.7-39), PV 0.4 (p = 0.4, 95% CI: 0.04-3.2), ET-1.6 (p = 0.7, 95% CI: 0.43-6.2) and patients with co-existent MPS-2.8 (p=0.07, 95% CI: 0.91-8.6). However, after adjusting for age, sex, dyslipidemia, diabetes, hypertension and tobacco use, the difference in the prevalence of CAD between the various categories of MPS was not significant. CONCLUSION: Contrary to conventional belief, we did not find an increased prevalence of CAD in patients with either PV or ET. In fact, patients with MF had a significantly higher prevalence of CAD. However, this difference appears to be due to the increased age at diagnosis of MF. The conventional risk factors for CAD appear to be the major determinants of CAD among patients with MPS. PMID- 12893022 TI - The 2756A>G variant in the gene encoding methionine synthase: its relation with plasma homocysteine levels and risk of coronary heart disease in a Dutch case control study. AB - INTRODUCTION: Elevated plasma homocysteine levels have been associated with increased risk of cardiovascular disease. A 2756A>G polymorphism has been found in the gene (MTR) coding for methionine synthase, an enzyme catalyzing remethylation of homocysteine to methionine. MATERIALS AND METHODS: In a Dutch case-control study comprising 123 cases with coronary heart disease (CHD) and 540 controls, we evaluated whether the MTR 2756A>G polymorphism was associated with plasma homocysteine, vitamin B12, folate concentrations, and CHD risk. RESULTS AND CONCLUSIONS: The polymorphism was not associated with fasting or post methionine load homocysteine concentrations. Individuals with the GG genotype had 30% lower vitamin B12 concentrations than individuals with AA or AG genotype (P < 0.05). After adjustment for CHD risk factors, the odds ratio (OR) of CHD was 4.0 (95% CI 1.4-11.6) for the GG genotype and 0.7 (95% CI 0.4-1.2) for the AG genotype, when compared to the AA genotype. In conclusion, despite the absence of an association with plasma homocysteine, the GG genotype represented a four-fold increased risk of CHD when compared to the AA genotype. Before putting effort in additional epidemiological studies, it needs to be established first whether this polymorphism has functional consequences for enzyme activity. PMID- 12893023 TI - Endothelial markers and HIV infection in the era of highly active antiretroviral treatment. AB - Many circumstances can induce activation and/or injury of the endothelium that plays a role in the development of vascular complications. Raised plasma levels of endothelial markers such as von Willebrand factor (vWF), soluble thrombomodulin (sTM) and soluble vascular cell adhesion molecule-1 (sVCAM-1) have a prognostic and/or diagnostic value. Human immunodeficiency virus-infected patients (HIV+) have a clustering of conditions that activate or injure the endothelium. Highly active antiretroviral treatment produces adverse effects such as dyslipemia, insulin resistance (IR) and body fat changes (named lipodystrophy syndrome) which may contribute to aggravate their endothelial perturbation. The aim of this study was to measure lipid profile, insulin resistance status, and endothelial markers in 38 HIV+ naive of antiretroviral treatment and 63 HIV+ under highly active antiretroviral treatment (33 with lipodystrophy syndrome and 30 without it). Body fat distribution was also evaluated by dual-energy X-ray absorptiometry (DEXA) analysis. Thirty-one HIV negative subjects were used as controls. We looked for association between variables. Insulin resistance status was a common finding in the four groups. Lipodystrophic patients presented an atherothrombotic lipid profile [elevated levels of triglycerides (TG), low density lipoprotein cholesterol (LDL-chol) and apolipoprotein-B (APO-B)] and a strong loss of fat in legs and arms (lipoatrophy). All endothelial markers evaluated in our naive patients were higher as compared to control group. sVCAM-1 in HIV+ under therapy without lipodystrophy syndrome showed significantly decreased levels as compared to naive group (487 vs. 666 ng/ml) and vWF and sTM tended to diminish although they did not show a significant difference (130% vs. 170%, 41 vs. 45 ng/ml, respectively). Lipodystrophic patients showed a tendency to increased levels of endothelial activation markers (sVCAM-1: 500 ng/ml and vWF: 154%) together with significantly increased levels of an endothelial injury marker (sTM: 50 ng/ml) with respect to HIV+ under therapy without lipodystrophy syndrome. Plasma levels of sTM, as an endothelial injury marker, correlated with peripheral lipoatrophy (rho = -0.357) in lipodystrophic patients. In conclusion, despite the beneficial immunology effect of highly active antiretroviral treatment and the apparent decrease in the endothelial perturbation, the patients who develop lipodystrophy present altered endothelial markers and other risk factors, such as IR and dyslipemia, which turn them into a high atherothrombotic risk group. PMID- 12893024 TI - Differential effect of the GPIIb/IIIa antagonist orbofiban on human platelet aggregate formation in vitro. AB - Platelet microaggregates have been demonstrated in the systemic circulation of patients with atherosclerotic cardiovascular diseases. Glycoprotein IIb/IIIa antagonists have been reported to play roles in platelet activation, which may be associated with pro-thrombotic events. We report the effect of the orally active GPIIb/IIIa antagonist, orbofiban, on human platelet microaggregate formation in vitro. Laser light scatter (LSS) technology was used to monitor small, medium and large platelet aggregates formed in platelet-rich plasma in response to ADP or thrombin receptor activating peptide (TRAP)-6. ADP at 0.5 microM induced only small platelet aggregates that were prevented by orbofiban in a concentration dependent manner. Likewise, orbofiban dissolved small aggregates after their formation. In marked contrast, in the presence of strong agonist stimulation (20 microM ADP or 3 microM TRAP-6) which generated primarily large platelet aggregates, orbofiban blocked the large aggregates while significantly augmenting small aggregates by three- to sixfold. The data suggest that GPIIb/IIIa antagonists do not induce platelet microaggregation directly but may augment small platelet microaggregate formation indirectly at conditions of strong stimuli. The percentage of the microaggregate population expressing P-selectin remained the same in the presence of orbofiban, indicating that these small microaggregates remain activated, although the mean intensity of expression was diminished, possibly reflecting the reduced size of the particles or density of P selectin molecules. In summary, an increase in small platelet microaggregates might have contributed to pro-thrombotic events in orbofiban-treated patients. PMID- 12893025 TI - Isosorbide-5-mononitrate treatment prevents cyclosporin A-induced platelet hyperactivation and the underlying nitric oxide-cyclic guanosine-3',5' monophosphate disturbances. AB - INTRODUCTION: The clinical use of cyclosporin A (CsA) is commonly associated with the development of hypertension and increased risk of thromboembolic events. Decreased endothelium-dependent relaxation and increased platelet activation seems to be involved on those side effects, but the underlying mechanisms are not yet elucidated. The present study aimed to evaluate the CsA effect on the platelet NO-cyclic guanosine-3',5'-monophosphate (cGMP) pathway and the putative benefits of concomitant isosorbide-5-mononitrate (IS-5-MN) administration on CsA induced hypertension and on platelet hyperactivation. MATERIALS AND METHODS: Blood pressures, platelet NO synthase activity and cGMP content, intracellular free calcium concentration ([Ca2+]i) and whole blood platelet aggregation were assessed in three rat groups orally treated, during 7 weeks, with the following diets: orange juice (control group), 5 mg/kg/day of CsA (CsA group) and 150 mg/kg/day, b.i.d., of IS-5-MN for 2 weeks and IS-5-MN plus 5 mg/kg/day of CsA for 7 weeks (IS-5-MN+CsA group). RESULTS: IS-5-MN treatment has prevented hypertension development obtained in the solely CsA-treated rats. CsA treatment has inhibited NOS activity, which was reverted by the concomitant IS-5-MN and CsA administration. On the contrary, platelets from CsA-treated rats had cGMP content increased when compared with the control rats. The variation obtained when ISMN was present was less predominant. Therefore, the organic nitrate treatment has prevented platelet hyperactivation, namely, by decreasing thrombin-evoked [Ca2+]i and collagen-evoked platelet aggregation, when compared with the solely CsA treated group. The preventive effect of IS-5-MN was reinforced by electron microscopy studies of platelet activation. CONCLUSIONS: By increasing [Ca2+]i and aggregation, CsA induces platelet hyperactivation and simultaneously increases cGMP content, which might represent a compensatory inhibitory mechanism. The concomitant IS-5-MN treatment prevents the above-mentioned platelet hyperreactivity and tends to normalize the NO-cGMP pathway as well as the development of hypertension. PMID- 12893026 TI - Role of ascorbic acid in the modulation of inhibition of platelet aggregation by polymorphonuclear leukocytes. AB - OBJECTIVES: We investigated the modulatory effect of ascorbate on the inhibition of platelet aggregation response by polymorphonuclear leukocytes (PMNs) and characterized the mechanism of the inhibitory response. BACKGROUND: PMNs have been reported to play a significant role in vascular homeostasis by releasing various factors including short-lived reactive oxygen species (ROS) and nitric oxide (NO). NO prevents the activation of circulating platelets and plays a significant role in hemostasis. In addition, PMNs also have the capacity to store very high concentrations of ascorbate. The physiological implications of storing such high concentrations of an antioxidant by a cell-releasing free radicals is unknown, viz. a viz. hemostatic regulation. METHODS: ADP-induced aggregation in human, monkey and rat platelet-rich plasma (PRP) was monitored in the presence of PMNs treated with varying concentrations of ascorbate/dehydroascorbate. NO generation from rat and human PMNs treated with ascorbate was monitored on a FACS Calibur flow cytometer and intraplatelet cyclic guanosine 3',5'-monophosphate (cGMP) levels was also measured. RESULTS: PMNs induced a cell number and time dependent inhibition of ADP-induced aggregation. The PMNs dependent inhibition was enhanced significantly at 30 min by ascorbate (300 microM). Ascorbate seemed to exert its effects through its oxidized product, dehydroascorbate, as the effects was prevented in the presence of D-glucose (10 mM). Dehydroascorbate elicited significant potentiation of the PMNs induced inhibitory responses and these effects were mediated by the release of NO and subsequent activation of platelet guanylyl cyclase. Flow cytometry experiments with human and rat PMNs confirmed the release of NO and the elevated platelet cGMP levels confirmed NO mediated activation of guanylyl cyclase. CONCLUSIONS: Ascorbate in circulation seems to prevent the activation of platelets by enhancing the release of antiaggregatory NO, from neighbouring or cohabitant PMNs. The ascorbate effect is mediated through its conversion to dehydroascorbate, subsequently, gets taken up by the cell and converted back to ascorbate. Intracellular ascorbate potentiates the release of NO from the PMNs and subsequently activates guanylyl cyclase in the platelets. PMID- 12893027 TI - Anti-thrombotic activity of PDR, a newly synthesized L-Arg derivative, on three thrombosis models in rats. AB - The possibility of a newly synthesized L-arginine derivative, polyaspartoyl-L arginine (PDR), as a novel anti-thrombotic agent and its mode of action were investigated. The anti-platelet effects of PDR in rats ex vivo, anti-thrombotic effects in three thrombosis models in rats and its effect on some autacoids (nitric oxide [NO], thromboxane [TXA2] and prostacyclin [PGI2]) were studied. PDR (i.g.) significantly inhibited ADP-, collagen- or thrombin-induced rat platelet aggregation. In arteriovenous shunt model and ferric chloride-induced arterial thrombosis model in rats, PDR (i.g.) significantly reduced the thrombus weight. In electrical stimulation-induced arterial thrombosis in rats, PDR (i.v.) dose dependently prolonged the thrombus occlusion time (OT). PDR increased the concentration of NO in plasma. In contrast with aspirin (ASA), PDR did not influence on the TXA2 and PGI2 levels in plasma. In conclusion, PDR is provided with significant inhibitory effect on platelet aggregation and prevention effect on platelet related thrombosis, which is probably attributed to its inhibition on platelet function by L-arginine-NO pathway. The results demonstrate that PDR is a novel, oral and venous effective platelet aggregation inhibitor and has a possibility used as an anti-thrombotic agent. PMID- 12893028 TI - Effects of armagnac extracts on human platelet function in vitro and on rat arteriovenous shunt thrombosis in vivo. AB - INTRODUCTION: The "French paradox", a low cardiovascular mortality compared to the prevalent risk factors, has been attributed to the regular use of red wine, and to the polyphenols it contains. These have among other effects an antioxidant and antithrombotic effect. The French paradox is maximal in southwest France, a region which is the region of production of armagnac, an oak cask aged spirit also rich in polyphenols. METHOD: We tested the effects of a freeze-dried extract of 12-year-old armagnac (EA88) on in vitro human platelet adhesion, and on aggregation induced by collagen or ADP, in the presence or absence of hypoxanthine-xanthine oxidase (HX/XO), at concentrations ranging from 5 x 10(-9) to 5 x 10(-3) g/l, after 15-60 min incubation. We also tested the effects of 2 week oral treatment with 1, 5 and 25 mg/kg EA88 in a rat arteriovenous shunt thrombosis model. RESULTS: EA88 inhibited ADP-induced but not collagen-induced human platelet aggregation in vitro in a concentration- and incubation time dependent manner, which was greater in the presence of HX/XO. In vivo, giving rats a daily oral dose of EA88 for 2 weeks inhibited thrombus formation in a dose dependent manner, for doses consistent with the habitual human use of armagnac. CONCLUSION: Armagnac extract EA88 had an antiplatelet and antithrombotic effect that if confirmed in man could contribute to explain the intensity of the French paradox in southwest France. PMID- 12893029 TI - Tissue factor pathway inhibitor production by human proximal tubular epithelial cells in culture. AB - Fibrin deposition in the peritubular capillaries and along the tubular basement membrane is commonly observed in several renal diseases and suggests the involvement of blood coagulation in tubulointerstitial damage. It has been demonstrated that tissue factor (TF) is present in tubular epithelial cells of animal models of nephritis. Tissue factor pathway inhibitor (TFPI) regulates the extrinsic pathway of blood coagulation through its ability to inhibit TF activity and it is now thought to be produced mainly by the vascular endothelial cells. We examined whether human proximal tubular epithelial cells (PTEC) could produce TFPI and attempted to clarify the regulatory factors affecting TFPI production. Cultured human PTEC were used. The procoagulant activity (PCA) in PTEC lysate was quantified by measurement of the one-stage recalcification time. TFPI in the cell supernatants was measured by ELISA. The mRNA of TF and TFPI in PTEC was analyzed by reverse transcriptase polymerase chain reaction (RT-PCR). PCA which is compatible with TF activity was present in the PTEC lysate. TF mRNA and TFPI mRNA were detected in PTEC. The amount of TFPI increased over time in the cell supernatants. Immnoblot analysis revealed 40 kD protein of TFPI, and TFPI antigen was demonstrated in PTEC by immunofluorescence. The concentration of TFPI was significantly increased following incubation with thrombin and heparin in a dose- and time-dependent manner, although the amount of TFPI mRNA was not changed. Our study showed that TFPI is produced in cultured PTEC and added one more cell type that produced TFPI other than endothelial cells. Thrombin and heparin stimulated TFPI secretion from PTEC. TFPI of PTEC may act against generation of thrombin and tubular fibrin formation induced by tissue factor activation. The augmentation of TFPI secretion by heparin may play an important role in the modulation of anticoagulant properties of PTEC. PMID- 12893030 TI - Noninvasive transcutaneous ultrasound augments thrombolysis in the left circumflex coronary artery--an in vivo canine study. AB - BACKGROUND: Ultrasound has the potential to augment chemical thrombolysis. METHODS AND RESULTS: Thrombotic occlusions in the left circumflex artery (LCx) were induced in 27 dogs. Sixty minutes later, tissue-type plasminogen activator (t-PA) was given intravenously over 90 min. Thrombotic occlusions (n = 20) were treated with concomitant transcutaneous low frequency (27 kHz), continuous wave (CW) (n = 10) or pulsed wave (PW) (n = 10) ultrasound. Tissue-type plasminogen activator plus ultrasound (n = 20) vs. tissue-type plasminogen activator alone (n=7) resulted in more frequent Thrombolysis in Myocardial Infarction (TIMI) 3 flow (90% vs. 43%, P = 0.024) and less reocclusion (11% vs. 67%, P = 0.080). At 60 min, median TIMI grade flow for tissue-type plasminogen activator alone was 2 (mean: 1.43 +/- 1.40) compared to 3 (mean: 2.70 +/- 0.95) for tissue-type plasminogen activator plus continuous as well as pulsed wave ultrasound (P = 0.035). Continuous wave and pulsed wave ultrasound were equally effective in augmenting thrombolysis. Histologically, no ultrasound-mediated injury to the myocardium or coronary arteries occurred. CONCLUSION: Both transcutaneous low frequency continuous wave ultrasound and pulsed wave ultrasound enhance tissue type plasminogen activator-mediated thrombolysis of the posterior circulation with higher TIMI 3 flow rates and less reocclusion than with tissue-type plasminogen activator alone. In addition, at the energy levels used, low frequency ultrasound appears safe. PMID- 12893031 TI - Urine and plasma levels of fibrinopeptide B in patients with deep vein thrombosis and pulmonary embolism. AB - BACKGROUND: Many patients with pulmonary thromboembolism remain undiagnosed, possibly because of the difficulty clinicians have in determining which patients merit work-up with accurate (but expensive) imaging techniques. OBJECTIVES: We present the first prospective clinical study of pulmonary embolism (PE) and deep venous thrombosis (DVT) detection using the FPBtot assay, which measures fibrinopeptide B and its first derivative, des-arginine fibrinopeptide B. METHODS: Twenty three patients with signs or symptoms of PE or DVT were enrolled in the study prior to the performance of definitive testing. Using a novel immunoassay, FPBtot levels were measured in urine and plasma samples from patients as well as from healthy controls. Urine and plasma FPBtot levels were compared to the diagnostic results, as blindly adjudicated by one of the investigators. Patients were excluded if they withdrew (n =1), had inconclusive diagnostic testing (n = 7), or did not give samples (n = 2 for urine, n = 3 for plasma). RESULTS: The mean FPBtot concentration in the urine of the 'DVT/PE positive' group was 78.4 +/- 35.2 ng/ml and 2.7 +/- 1.9 ng/ml in the 'DVT/PE negative' group (p = 0.03). The urine FPB(tot) concentrations in the 'DVT/PE negative' group were not significantly different from those in the healthy control group (2.2 +/- 0.4 ng/ml, p = 0.40). The area under the ROC curve for urine FPB(tot) concentrations was 97.3 +/- 3.8%, suggesting a high degree of diagnostic accuracy. Plasma FPB(tot) concentrations were not significantly different between groups. CONCLUSIONS: Urine FPBtot levels may help detect patients with PE and DVT. PMID- 12893032 TI - Size and power required for motion with implication for the evolution of early hominids. AB - The fossil record of early hominids (early human ancestors) suggests that their stature and weight had a tendency to increase, but their robusticity (the proportion of radius to length) to decrease. Using a simple musculo-skeletal model, this paper explores possible relationships between size, power required for motion (PRM) and cycle-time, deriving relationships which indicate that PRM per unit of mass and velocity is proportional to robusticity, but inversely proportional to stature. The results derived appear to be in general agreement with published data from physiological experiments. If the material properties of early hominids were similar to those of modern humans and the achievement of minimum PRM was the selective criterion, human stature might tend to increase slightly in human evolution (and, if selective pressures are not removed, might do so in the future but at lower rate). If mobility and stability under loading are the selective criteria, however, human size should not substantially increase in the future. PMID- 12893033 TI - The implant thread as a retention element in cortical bone: the effect of thread size and thread profile: a finite element study. AB - Assuming that high stress peaks in the bone can trigger bone resorption a screw shaped bone implant should be given such a design that the peak stresses arising in the bone, as a result of a certain load, are minimized. Using idealized assumptions the aim of the study was to analyse the effect of variations of the size and the profile of the thread of an axially loaded, screw-shaped, bone implant upon the magnitude of the stress peaks in cortical bone. The investigation was performed by means of axisymmetric finite element analysis. It was found that the shape of the thread profile has a profound effect upon the magnitude of the stresses in the bone and that very small threads of a favourable profile can be quite effective. PMID- 12893034 TI - Ultrasound indentation of bovine knee articular cartilage in situ. AB - We have earlier developed a handheld ultrasound indentation instrument for the diagnosis of articular cartilage degeneration. In ultrasound indentation, cartilage is compressed with the ultrasound transducer. Tissue thickness and deformation are calculated from the A-mode ultrasound signal and the stress applied is registered with the strain gauges. In this study, the applicability of the ultrasound indentation instrument to quantify site-dependent variation in the mechano-acoustic properties of bovine knee cartilage was investigated. Osteochondral blocks (n=6 per site) were prepared from the femoral medial condyle (FMC), the lateral facet of the patello-femoral groove (LPG) and the medial tibial plateau (MTP). Cartilage stiffness (dynamic modulus, E(dyn)), as obtained with the ultrasound indentation instrument in situ, correlated highly linearly (r=0.913, p<0.01) with the values obtained using the reference material-testing device in vitro. Reproducibility (standardized coefficient of variation) of the ultrasound indentation measurements was 5.2%, 1.7% and 3.1% for E(dyn), ultrasound reflection coefficient of articular surface (R) and thickness, respectively. E(dyn) and R were site dependent (p<0.05, Kruskall-Wallis H test). E(dyn) was significantly higher (p<0.05, Kruskall-Wallis Post Hoc test) in LPG (mean+/-SD: 10.1+/-3.1MPa) than in MTP (2.9+/-1.4MPa). In FMC, E(dyn) was 4.6+/ 1.3MPa. R was significantly (p<0.05) lower at MTP (2.0+/-0.7%) than at other sites (FMC: 4.2+/-0.9%; LPG: 4.4+/-0.8%). Cartilage glycosaminoglycan concentration, as quantified with the digital densitometry, correlated positively with E(dyn) (r=0.678, p<0.01) and especially with the equilibrium Young's modulus (reference device, r=0.874, p<0.01) but it was not associated with R (r=0.294, p=0.24). We conclude that manual measurements are reproducible and the instrument may be used for detection of cartilage quality in situ. Especially, combined measurement of thickness, E(dyn) and R provides valuable diagnostic information on cartilage status. PMID- 12893035 TI - Cavitation phenomena in mechanical heart valves: the role of squeeze flow velocity and contact area on cavitation initiation between two impinging rods. AB - In this study, the closing dynamics of two impinging rods were experimentally analyzed to simulate the cavitation phenomena associated with mechanical heart valve closure. The purpose of this study was to investigate the cavitation phenomena with respect to squeeze flow between two impinging surfaces and the parameter that influences cavitation inception. High-speed flow imaging was employed to visualize and identify regions of cavitation. The images obtained favored squeeze flow as an important mechanism in cavitation inception. A correlation study of the effects of impact velocities, contact areas and squeeze flow velocity on cavitation inception showed that increasing impact velocities results in an increase in the risk of cavitation. It was also shown that for similar impact velocities, regions near the point of impact were found to cavitate later for those with smaller contact areas. It was found that the decrease in contact areas and squeeze flow velocities would delay the onset and reduce the intensity of cavitation. It is also interesting to note that the squeeze flow velocity alone does not provide an indication if cavitation inception will occur. This is corroborated by the wide range of published critical squeeze flow velocity required for cavitation inception. It should be noted that the temporal acceleration of fluid, often neglected in the literature, can also play an important role on cavitation inception for unsteady flow phenomenon. This is especially true in mechanical heart valves, where for the same leaflet closing velocity, valves with a seat stop were observed to cavitate earlier. Based on these results, important inferences may be made to the design of mechanical heart valves with regards to cavitation inception. PMID- 12893036 TI - Gender dependent cervical spine segmental kinematics during whiplash. AB - Clinical and epidemiological studies have frequently reported that female occupants sustain whiplash injuries more often than males. The current study was based on the hypothesis that segmental level-by-level cervical intervertebral motions in females are greater than in males during rear impact. The hypothesis was tested by subjecting 10 intact human cadaver head-neck complexes (five males, five females) to rear impact loading. Intervertebral kinematics were analyzed as a function of spinal level at the time of maximum cervical S-curve, which occurred during the loading phase. Segmental angles were significantly greater (p<0.05) in female specimens at C2-C3, C4-C5, C5-C6, and C6-C7 levels. Because greater angulations are associated with stretch in the innervated components of the cervical spinal column, these findings may offer a biomechanical explanation for the higher incidence of whiplash-related complaints in female patients secondary to rear impact acceleration. PMID- 12893037 TI - A method to estimate in vivo dynamic articular surface interaction. AB - This paper describes a method to calculate and visualize the proximity of subchondral bone surfaces during dynamic movement. This method combines high speed biplane radiographic image data and three-dimensional (3D) bone surface information derived from computed tomography to determine subchondral bone motion during dynamic activities. Knowledge of in vivo subchondral bone motion may be useful in the study of osteoarthritis, in biomechanical modeling, and in identifying normal and pathological joint mechanics. This method can be used to identify the regions of close contact during dynamic motion, to calculate the surface area of subchondral bone within close contact, and to determine the changing position of the close contact area during dynamic activities. None of this informations can be obtained using other currently available 3D motion analysis techniques. Example applications showing dynamic in vivo tibio-femoral bone surface motion during canine gait and human one-legged hopping are presented. PMID- 12893038 TI - Instant center of rotation estimation using the Reuleaux technique and a Lateral Extrapolation technique. AB - A mathematical model of a rolling wheel was used to investigate the errors encountered when the Reuleaux technique is employed to estimate planar instant centers of rotation (ICRs). The investigation showed that large errors can result when this pole measurement technique is applied to objects rotating more than 12 degrees. The investigation also showed that these errors can be substantially reduced by applying a new Lateral Extrapolation technique to the pole data. When the Reuleaux technique is applied to marks on a 10cm radius wheel, the resulting offset errors from the ICR are 3.96cm for a 45 degrees roll and 1cm for a 12 degrees roll. Following lateral extrapolation, these offset errors reduce to 0.52cm for the 45 degrees roll and less than 0.04cm for the 12 degrees roll. Thus, the extrapolation technique is over seven times more accurate for a 45 degrees roll, and over 25 times more accurate for a 12 degrees roll. The extrapolation technique has been validated with the model for joints that exhibit both slip and roll, such as the knee. As joint ICR pathway measurement can be used to detect pathology, these accuracy improvements offer potential benefits for clinical applications. PMID- 12893039 TI - Stretch-induced, steady-state force enhancement in single skeletal muscle fibers exceeds the isometric force at optimum fiber length. AB - Stretch-induced force enhancement has been observed in a variety of muscle preparations and on structural levels ranging from single fibers to in vivo human muscles. It is a well-accepted property of skeletal muscle. However, the mechanism causing force enhancement has not been elucidated, although the sarcomere-length non-uniformity theory has received wide support. The purpose of this paper was to re-investigate stretch-induced force enhancement in frog single fibers by testing specific hypotheses arising from the sarcomere-length non uniformity theory. Single fibers dissected from frog tibialis anterior (TA) and lumbricals (n=12 and 22, respectively) were mounted in an experimental chamber with physiological Ringer's solution (pH=7.5) between a force transducer and a servomotor length controller. The tetantic force-length relationship was determined. Isometric reference forces were determined at optimum length (corresponding to the maximal, active, isometric force), and at the initial and final lengths of the stretch experiments. Stretch experiments were performed on the descending limb of the force-length relationship after maximal tetanic force was reached. Stretches of 2.5-10% (TA) and 5-15% lumbricals of fiber length were performed at 0.1-1.5 fiber lengths/s. The stretch-induced, steady-state, active isometric force was always equal or greater than the purely isometric force at the muscle length from which the stretch was initiated. Moreover, for stretches of 5% fiber length or greater, and initiated near the optimum length of the fiber, the stretch-enhanced active force always exceeded the maximal active isometric force at optimum length. Finally, we observed a stretch-induced enhancement of passive force. We conclude from these results that the sarcomere length non-uniformity theory alone cannot explain the observed force enhancement, and that part of the force enhancement is associated with a passive force that is substantially greater after active compared to passive muscle stretch. PMID- 12893040 TI - Focal cryogen insults for inducing segmental osteonecrosis: computational and experimental assessments of thermal fields. AB - Orthopaedic management of femoral head osteonecrosis is a common clinical problem for which there presently is no good solution. Current animal models are inappropriate to study potential new solutions, since it has been difficult to replicate the natural history of structural collapse seen in the human disorder. Recently, progression to collapse was obtained for cryogenically induced osteonecrosis in emus, although the lesions involved were imprecisely controlled in terms of size or location. A new cryo-insult probe is here reported for the purpose of delivering well-prescribed local thermal insults in this new animal model, while minimizing damage to non-targeted regions. Finite element analysis was used to elucidate the influence of operator-controlled parameters upon the temporal/spatial variation of the thermal field. The numerical formulation includes convective heat transfer attributable to tissue bed perfusion. The computational results agreed closely with the results of thermocouple recordings in a companion bench-top experiment. The cryo-insult probe successfully produced segmental lesions in the emu model of sizes comparable to the computed freeze front diameters. PMID- 12893041 TI - Predicting the dynamic postural control response from quiet-stance behavior in elderly adults. AB - Human postural sway, as measured by fluctuations of the center of pressure (COP) under the feet of a quietly standing individual, can be characterized as a stochastic process. The fluctuation-dissipation theorem (FDT) provides a linear relationship between the fluctuations of a quasi-static, stochastic system to the same system's relaxation to equilibrium following a perturbation. We applied a similar linear relationship, based on the FDT, to the human postural control system to explore whether anterior-posterior (AP) fluctuations of the COP during quiet stance can be used to predict the AP response of the postural control system to a weak posteriorly directed mechanical perturbation (tug or pull at the waist). We tested 10 healthy elderly (mean age of 69yr) and 10 healthy young (mean age of 25yr) adult subjects. We found that this linear relationship was applicable to the postural control system of all 10 young and eight of the 10 elderly adult subjects. These results suggest that it is possible to predict an individual's dynamic response to a mild perturbation using quiet-stance data, regardless of age. The existence of this FDT-based linear relationship with respect to the human postural control system suggests that, for a given individual, the postural control system may use the same control mechanisms during quiet stance and mild-perturbation conditions, regardless of age. PMID- 12893042 TI - Influence of body segment parameters and modeling assumptions on the estimate of center of mass trajectory. AB - This study sought to determine the effect of inaccuracies in body segment parameters and modeling assumptions on the estimate of antero-posterior center of mass (COM) trajectory. Four different methods, one based on segmental kinematics, and three methods based on kinetic recordings were compared via simulation. Kinematic patterns (quiet stance, ankle-related sway, hip-ankle-related sway, sit up and sit-up-sit-down) were tested with a 2D four-link model of the body and the ground reaction force vector was obtained by inverse dynamics. Errors in the estimation of body segment parameters were simulated by applying a +/-10% variation to one or more parameters at a time. These errors propagated differently to the COM estimated location between methods, between parameters within the same method, and between tasks. The kinematics-based method was the most sensitive to body segment parameters, with special regards to segment lengths and head-arms-trunk parameters. Root mean square error between estimated and simulated COM location reached 19mm in balance-related tasks and 38.3mm in sit-up-sit-down. The kinetics-based methods were largely less sensitive to inaccuracies in body segment parameters. In particular, the technique proposed by Zatsiorsky and King (J. Biomech. 31 (1998) 161), was completely insensitive to segment parameters. On the other hand the kinetics-based methods showed an intrinsic estimation error, due to the underlying model assumptions. The methods based on the double integration of horizontal force had better outcomes with tasks challenging such assumptions, with a maximal error in COM location of 15mm in the sit-up-sit-down. The method proposed by Shimba (J. Biomech. 17 (1984) 53) showed the best trade-off between sensitivity to body segment parameters and estimation performances given the ideal test conditions. PMID- 12893043 TI - A rate-dependent microcrack-bridging model that can explain the strain rate dependency of cortical bone apparent yield strength. AB - Although there are empirical correlations between strain rate, cortical and cancellous bone apparent stiffness, apparent yield strength, apparent ultimate strength and cortical bone fracture toughness, a mechanistic description for these phenomena is lacking. Microcracking is a major mechanism in cortical and cancellous bone failure, however, microdamage content alone cannot explain the strain rate dependence of bone strength without considering time-dependent behavior of the crack. Using a rate-dependent model of a fiber-bridged microcrack and data from the literature, we demonstrate that the experimental apparent yield strength of bone can be predicted directly from measurements of apparent moduli of elasticity of bone constituents and failure strain of the collagenous matrix. Yield strength predictions for estrogen depleted bone were made using the model and data from ovariectomized sheep. It was predicted that the yield strength of estrogen-deficient bone is comparable to that of normal bone within strain rates associated with physiological activities. For high strain rates, however, the strength of estrogen-depleted bone was predicted to be much weaker than normals suggesting a higher fracture risk due to impact from falls, for individuals with estrogen-depleted bones such as in post-menopausal osteoporosis. PMID- 12893044 TI - Obesity is not associated with increased knee joint torque and power during level walking. AB - While it is widely speculated that obesity causes increased loads on the knee leading to joint degeneration, this concept is untested. The purpose of the study was to identify the effects of obesity on lower extremity joint kinetics and energetics during walking. Twenty-one obese adults were tested at self-selected (1.29m/s) and standard speeds (1.50m/s) and 18 lean adults were tested at the standard speed. Motion analysis and force platform data were combined to calculate joint torques and powers during the stance phase of walking. Obese participants were more erect with 12% less knee flexion and 11% more ankle plantarflexion in self-selected compared to standard speeds (both p<0.02). Obese participants were still more erect than lean adults with approximately 6 degrees more extension at all joints (p<0.05, for each joint) at the standard speed. Knee and ankle torques were 17% and 11% higher (p<0.034 and p<0.041) and negative knee work and positive ankle work were 68% and 11% higher (p<0.000 and p<0.048) in obese participants at the standard speed compared to the slower speed. Joint torques and powers were statistically identical at the hip and knee but were 88% and 61% higher (both p<0.000) at the ankle in obese compared to lean participants at the standard speed. Obese participants used altered gait biomechanics and despite their greater weight, they had less knee torque and power at their self selected walking speed and equal knee torque and power while walking at the same speed as lean individuals. We propose that the ability to reorganize neuromuscular function during gait may enable some obese individuals to maintain skeletal health of the knee joint and this ability may also be a more accurate risk indicator for knee osteoarthritis than body weight. PMID- 12893045 TI - Mechanosensitivity of bone cells to oscillating fluid flow induced shear stress may be modulated by chemotransport. AB - Fluid flow has been shown to be a potent physical stimulus in the regulation of bone cell metabolism. In addition to membrane shear stress, loading-induced fluid flow will enhance chemotransport due to convection or mass transport thereby affecting the biochemical environment surrounding the cell. This study investigated the role of oscillating fluid flow induced shear stress and chemotransport in cellular mechanotransduction mechanisms in bone. Intracellular calcium mobilization and prostaglandin E(2) (PGE(2)) production were studied with varying levels of shear stress and chemotransport. In this study MC3T3-E1 cells responded to oscillating fluid flow with both an increase in intracellular calcium concentration ([Ca(2+)](i)) and an increase in PGE(2) production. These fluid flow induced responses were modulated by chemotransport. The percentage of cells responding with an [Ca(2+)](i) oscillation increased with increasing flow rate, as did the production of PGE(2). In addition, depriving the cells of nutrients during fluid flow resulted in an inhibition of both [Ca(2+)](i) mobilization and PGE(2) production. These data suggest that depriving the cells of a yet to be determined biochemical factor in media affects the responsiveness of bone cells even at a constant peak shear stress. Chemotransport alone will not elicit a response, but it appears that sufficient nutrient supply or waste removal is needed for the response to oscillating fluid flow induced shear stress. PMID- 12893046 TI - Fibril reinforced poroelastic model predicts specifically mechanical behavior of normal, proteoglycan depleted and collagen degraded articular cartilage. AB - Degradation of collagen network and proteoglycan (PG) macromolecules are signs of articular cartilage degeneration. These changes impair cartilage mechanical function. Effects of collagen degradation and PG depletion on the time-dependent mechanical behavior of cartilage are different. In this study, numerical analyses, which take the compression-tension nonlinearity of the tissue into account, were carried out using a fibril reinforced poroelastic finite element model. The study aimed at improving our understanding of the stress-relaxation behavior of normal and degenerated cartilage in unconfined compression. PG and collagen degradations were simulated by decreasing the Young's modulus of the drained porous (nonfibrillar) matrix and the fibril network, respectively. Numerical analyses were compared to results from experimental tests with chondroitinase ABC (PG depletion) or collagenase (collagen degradation) digested samples. Fibril reinforced poroelastic model predicted the experimental behavior of cartilage after chondroitinase ABC digestion by a major decrease of the drained porous matrix modulus (-64+/-28%) and a minor decrease of the fibril network modulus (-11+/-9%). After collagenase digestion, in contrast, the numerical analyses predicted the experimental behavior of cartilage by a major decrease of the fibril network modulus (-69+/-5%) and a decrease of the drained porous matrix modulus (-44+/-18%). The reduction of the drained porous matrix modulus after collagenase digestion was consistent with the microscopically observed secondary PG loss from the tissue. The present results indicate that the fibril reinforced poroelastic model is able to predict specifically characteristic alterations in the stress-relaxation behavior of cartilage after enzymatic modifications of the tissue. We conclude that the compression-tension nonlinearity of the tissue is needed to capture realistically the mechanical behavior of normal and degenerated articular cartilage. PMID- 12893047 TI - The effect of roughness on biophysical stimuli at the bone-cartilage interface. AB - Numerical simulations are often used to investigate the effect of mechanical environment on fracture healing. Although these models exhibit biologically relevant mechanical parameters at the bone-callus interface, this interface is modelled as perfectly smooth when in fact it is rough. In this study, a macro micro-two-scale finite element model was used to determine if roughness significantly alters calculated local mechanical parameters. An idealized fracture healing poroelastic model with a small micro-modelled sub-domain of cartilaginous callus adjacent to rough bone was subjected to cyclic loading. The shear stress, tangential fluid velocity, and pore pressure were investigated. With roughness similar to that at the growth plate, solid matrix shear stress differed substantially with interface roughness, whereas interstitial fluid velocity and pore pressure were only slightly affected. Hence, when modelling local micro-mechanical environments near hard-soft tissue interfaces, interface roughness should be considered. PMID- 12893048 TI - Biomechanical properties of oesophagus wall under loading. AB - In this investigation, firstly, the biomechanical properties of different parts of oesophagus were determined. Oesophagus stress and strain are the greatest in the cervical part for all age groups. The human oesophagus deforms unevenly, depending on the direction of load in relation to the organ's axis, it exhibits anisotropical behaviour. With the age the values of mechanical parameters of the oesophagus wall reduce, in particular beginning from 45 years of age, but the modulus of elasticity increases. Biomechanical properties of the oesophagus depend on the architecture of its structure. By loading the organ in the circumferential direction, microfibrilae rupture and deformation of the muscular fibres occurs. With increase of load, collagenous fibres straighten and microruptures in collagenous fibrilae occur. With stretching of oesophagus longitudinally, collagenous fibres partially preserve their wavy and helical configuration. Therefore, higher resistance of the oesophageal wall occurs in the longitudinal direction. PMID- 12893049 TI - A novel method for measuring medial compartment pressures within the knee joint in-vivo. AB - A novel method for the measurement of knee joint forces in-vivo is described. A thin (0.2mm) flexible electronic pressure sensor was inserted through a narrow arthroscopic portal into the osteoarthritic medial compartment of the knee joint. The sensor partially covered the load bearing area. The surgery was performed under local anaesthetic during normal arthroscopic examination following patient consent. Results are presented for 11 patients. The method was used in a pilot study to assess the effects of four valgus knee braces on medial compartment forces. An analysis of variance could not detect un-loading by any brace although there were large variations in force output. These variations may be attributable to shifts in the sensor position. In-vivo measurement of joint force is technically feasible. PMID- 12893050 TI - A laser-aligned method for anthropometry of hands. AB - We have developed a laser-aligned measuring device to facilitate the measurement of linear hand dimensions. Our objective has been to make a simple, clinically applicable device for anthropometry of the hands in patients with arthritis in order to gauge the progression of hand deformities. The system provides an alternative to the use of rules, tapes, photogrammetry and least-squares devices which have previously been applied to hand measurements. The system delivers results which are not significantly different from those measured with rules and callipers and has a similar level of accuracy. The use of laser pointers for alignment facilitates measurements which necessitate picking the highest point of the hand. The device is portable and simple to use in the clinic. We have used it to measure hand dimensions in normal subjects and patients with arthritis. However, the system could equally well be used to facilitate measurement of other objects with an irregular surface profile. PMID- 12893051 TI - Statistical methods in finite element analysis by Dar FH, Meakin JR and Aspden RM J Biomech, 2002, Vol. 35, pp. 1155-1161. PMID- 12893052 TI - Spatial distribution of hip capsule structural and material properties. PMID- 12893053 TI - Measuring body segment parameters: X-ray versus gamma scanning. PMID- 12893054 TI - Cryptic clues as to how water-soluble protein toxins form pores in membranes. AB - Pore-forming protein toxins possess the remarkable property that they can exist either in a stable water-soluble state or as an integral membrane pore. In order to convert from the water-soluble to the membrane state, the toxin must undergo large conformational changes. Recent work on a class of pore-forming toxins that are rich in beta-sheet content suggests a common mechanism of membrane insertion may exist despite these toxins possessing very different primary, tertiary and quaternary structures. PMID- 12893055 TI - Complex yessotoxins profile in Protoceratium reticulatum from north-western Adriatic sea revealed by LC-MS analysis. AB - While the occurrence of yessotoxin (YTX) has been reported worldwide from Protoceratium reticulatum, the biogenetic origin of some YTX analogues is still unknown, thus raising an issue whether they are metabolites of YTX formed in mussels or true products of different dinoflagellate species. Findings reported herein suggest that P. reticulatum from the north-western Adriatic sea is responsible for production, together with YTX (1), of homoYTX (2), 45-OHYTX (3), carboxyYTX (5), and noroxoYTX (7). YTX and its analogues have been determined by high performance liquid chromatography coupled with electrospray ion trap mass spectrometry (HPLC-MS and HPLC-MSn experiments). The result is the first to confirm production of these YTX analogues from this species and indicate it as a producing organism of homoYTX, 45-OHYTX, carboxyYTX, and noroxoYTX found in shellfish. PMID- 12893056 TI - Huwentoxin-V, a novel insecticidal peptide toxin from the spider Selenocosmia huwena, and a natural mutant of the toxin: indicates the key amino acid residues related to the biological activity. AB - A neurotoxin peptide (named Huwentoxin-V) was purified from the venom of the Chinese bird spider Selenocosmia huwena by a combination of ion exchange chromatography and reverse phase HPLC. HWTX-V has 35 amino acid residues, and is in perfect agreement with the molecular mass 4111.4 Da identified by mass spectrometry. A natural mutant of the toxin (called mHuwentoxin-V) was also isolated from the venom. mHWTX-V was only truncated two amino acid residues from the C-terminus of HWTX-V, and its molecular weight is 3877.1 Da determined by mass spectrometry. The six cysteine residues in each sequence of the two peptides suggest three disulfide bridges, the present of which was demonstrated by mass spectrometry after dithiothreiotol reduce and S-carboxymethylation. The primary structure of the two toxins exhibits sequence identity with other spider toxins such as ProTx-I (64%), SGTx (57%), SNX-482 (55%), and Hanatoxin (54%). HWTX-V can reversibly paralyze locusts and cockroaches for several hours with a ED50 value as 16 +/- 5 microg/g to locusts, and a larger dose of the toxin can cause death. However, mHWTX-V shows no significant effect on locusts and cockroaches. The structure-activity relationship indicates that the residues Phe34 and Ser35 in the C-terminus of HWTX-V are the key residues of the biological activity. PMID- 12893057 TI - The purification and partial characterisation of two novel metalloproteinases from the venom of the West African carpet viper, Echis ocellatus. AB - Separation of previously uncharacterised Echis ocellatus venom by phenyl-Superose FPLC (Fast Liquid Protein Chromatography) yielded eight protein fractions. Three of these displayed high proteolytic activity when assayed by in vivo and in vitro assays (including enzyme linked immunosorbant assay), and were further separated using Superdex 75 and Mono-Q FPLC. This resulted in the purification of a non haemorrhagic 24 kDa metalloproteinase (EoVMP1, pI 7.0), and a haemorrhagic 56 kDa metalloproteinase (EoVMP2, pI 5.5). Following tryptic digest, short amino acid sequences of EoVMP1 and EoVMP2 were obtained using Edman degradation. Both sequences displayed homology when aligned with existing snake venom metalloproteinases (SVMPs). The strong homology observed among previously well characterised SVMPs suggests that principles governing the interaction of substrates and inhibitors are likely to be similar for EoVMP1, EoVMP2 and all members of the reprolysin family. PMID- 12893058 TI - Ostreopsis sp., a possible origin of palytoxin (PTX) in parrotfish Scarus ovifrons. AB - A clone of toxic dinoflagellate Ostreopsis sp. and six specimens of a parrotfish Scarus ovifrons were collected in October 1997 at Tokushima Prefecture, Japan. Ostreopsis sp. was cultured in ESM medium for 16 days, and after rearing the cell pellet (about 4.0x10(5) cells) was extracted with 50% methanol, partitioned between an aqueous layer and 1-butanol layer, and biochemically tested. Similarly, the crude toxin from S. ovifrons was extracted, and tested. The mice injected with each 1-butanol layer from Ostreopsis sp. and S. ovifrons showed the common symptoms of convulsion, drowsiness and collapse, and died within 48 h. The lethal potency of Ostreopsis sp. was calculated to be 1.0x10(-4) MU/cell. All specimens of S. ovifrons were found to be toxic, where the highest potency was determined as 2 MU/g in muscle of one specimen. After being injected with toxins, the serum creatine phosphokinase levels of mice were found to be elevated. Toxins from Ostreopsis sp. and S. ovifrons showed delayed haemolytic activity with mouse and human erythrocytes, which was inhibited by an anti-palytoxin (PTX) antibody antibody and ouabain. Toxins from Ostreopsis sp. and S. ovifrons thus resembled each other, and strongly suggested to be PTX or its akin substance. Additionally, a considerable number of adherent Ostreopsis sp. was found in the gut contents of S. ovifrons during the heavy occurrence of Ostreopsis sp. in October 1997 at Tokushima Prefecture. From the above results, it can be strongly postulated that the dinoflagellate Ostreopsis sp. is the origin of PTX which is sequestered by the parrotfish S. ovifrons through food chain. PMID- 12893059 TI - A new Pan African polyspecific antivenom developed in response to the antivenom crisis in Africa. AB - Currently there is a crisis in the supply of antivenom for treatment of snake bite in sub-Saharan Africa. Commercial pressures have resulted in the reduction or even cessation of production of antivenom by European manufacturers while continued production of antivenom in Africa has been threatened by the privatisation of the only remaining company based in Africa. As a consequence, there has been an increase in snake bite morbidity and mortality in many African countries. Two Latin American antivenom manufacturers have agreed to produce antivenom suitable for Africa, using venoms from the species which are of the greatest medical importance in sub-Saharan Africa. Preclinical in vivo assays of neutralising potency demonstrated that a new Pan African antivenom produced in Colombia compared favourably with the existing commercial monospecific and polyspecific antivenoms. This new antivenom, and a similar product being manufactured in Costa Rica, are now candidates for clinical testing at an appropriate site in Africa. PMID- 12893060 TI - A novel conotoxin inhibiting vertebrate voltage-sensitive potassium channels. AB - Toxins from cone snail (Conus species) venoms are multiple disulfide bonded peptides. Based on their pharmacological target (ion channels, receptors) and their disulfide pattern, they have been classified into several toxin families and superfamilies. Here, we report a new conotoxin, which is the first member of a structurally new superfamily of Conus peptides and the first conotoxin affecting vertebrate K+ channels. The new toxin, designated conotoxin ViTx, has been isolated from the venom of Conus virgo and comprises a single chain of 35 amino acids cross-linked by four disulfide bridges. Its amino acid sequence (SRCFPPGIYCTSYLPCCWGICCSTCRNVCHLRIGK) was partially determined by Edman degradation and deduced from the nucleotide sequence of the toxin cDNA. Nucleic acid sequencing also revealed a prepropeptide comprising 67 amino acid residues and demonstrated a posttranslational modification of the protein by releasing a six-residue peptide from the C-terminal. Voltage clamp studies on various ion channels indicated that the toxin inhibits the vertebrate K+ channels Kv1.1 and Kv1.3 but not Kv1.2. The chemically synthesized product exhibited the same physiological activity and identical molecular mass (3933.7 Da) as the native toxin. PMID- 12893061 TI - Structural and biological characterization of a crotapotin isoform isolated from Crotalus durissus cascavella venom. AB - Envenoming by Crotalus durissus subspecies leads to coagulation disorders, myotoxicity, neurotoxicity and acute renal failure. The most serious systemic alteration and primary cause of death after snakebite is acute renal failure. In this work, we isolated crotapotin, an acid component (Crtp) of crotoxin from Crotalus durissus cascavella venom and we investigated its bactericidal and pro inflammatory activities as well as its renal effects in rat isolated perfused kidneys. Crtp was bactericidal to the Gram-negative species Xanthomonas axonopodis pv. passiflorae, but was less effective against the Gram-positive Claribacteri ssp, probably because of differences in the cell wall composition. Crtp showed a high amino acid sequence homology with other Crtps described in the literature (around of 90%) and its A and B chains had high conserved regions corresponding to the calcium-binding loop, catalytic site and helix 3 of PLA2. The Crtp showed moderate pro-inflammatory activity and increased significantly the inflammation evoked by PLA2 when co-injected or co-incubated with PLA2. The renal parameters evaluated included the perfusion pressure (PP), renal vascular resistance (RVR), urinary flow (UF), glomerular filtration rate (GFR) and percent of sodium tubular transport (%TNa+). Crotapotin (5 microg/ml) significantly increased the PP and RVR, whereas the GFR, UF and %TNa+ were unaffected. These results suggest that crotoxin is the main venom component responsible for nephrotoxicity and crotapotin contributes little to this phenomenom. The biological and bactericidal actions of Crtp also suggest that this protein may have functions other than simply acting as a chaperone for PLA2. PMID- 12893062 TI - Receptor-mediated toxicity of pahutoxin, a marine trunkfish surfactant. AB - Pahutoxin (PHN, choline chloride ester of 3-acetoxypalmitic acid) is a natural fish-killing (ichthyotoxic) agent derived from the defensive secretions of trunkfish. In spite of its obvious structural resemblance to synthetic cationic long-chain quaternary ammonium detergents, we show that PHN's action does not rely on its surfactant properties and is in fact, receptor-mediated. The above conclusion is supported by the following data: 1. Ichthyotoxicity is not related to its detergency or surfactivity, as indicated by the fact that the lethal concentration is about 1.5 orders of magnitude below its critical micelle concentration value (69 microM) and its liposomal/seawater partition coefficient is low (62-85); 2. The trunkfish is tolerant to its own pahutoxin; 3. Ichthyotoxicity occurs only upon application to the surrounding water, suggesting the existence of externally located receptors; 4. The receptor hypothesis was supported by the aid of equilibrium saturation binding assays revealing the presence of specific binding sites to PHN on the fish gill membranes; 5. The PHN tolerant trunkfish was shown to be devoid of PHN-binding sites. Some chemo ecological, and environmental implications are discussed. PMID- 12893063 TI - Antivenom serotherapy and volume resuscitation partially improve peripheral organ ischemia in dogs injected with scorpion venom. AB - We tested the hypothesis that fluid resuscitation combined with antivenom serotherapy given after injection of scorpion venom may increase cardiac output (CO) and blood pressure (BP) and prevent the decline in bicarbonate, pH and gastric perfusion. Seventeen anesthetized, mechanically ventilated dogs were given 0.1 mg/kg i.v. venom of the scorpion Leiurus quinquestriatus. The dogs were randomized into three groups: six dogs were given venom alone; three dogs were given 6 ml of antivenom 1 minute before venom injection; eight dogs were given 6 ml of antivenom and 20 ml/kg of synthetic colloid solution, 20 min after venom injection. Parameters reflecting respiratory and circulatory functions were determined at baseline and 120 min after venom injection. Scorpion venom caused a decrease in CO, BP, pH and HCO3-. Gastric mucosal perfusion was severely affected as assessed by mucosal pH (pHi) and the gradient between mucosal and arterial pCO2 (delta pCO2). Antivenom given before venom injection prevented all the effects induced by the venom. Antivenom and fluid given 20 min after venom injection caused a marked increase in CO and BP, but had no effect on pH and HCO3 decline (compared with venom alone). Gastric perfusion slightly improved as the increase in delta pCO2 was attenuated. The combination therapy of antivenom and fluid in this dog model is superior to the therapy of each of them alone. The marked and long-standing improvement of CO is promising and may suggest improvement in HCO3- and pH with time. PMID- 12893064 TI - Injuries caused by scorpionfishes (Scorpaena plumieri Bloch, 1789 and Scorpaena brasiliensis Cuvier, 1829) in the Southwestern Atlantic Ocean (Brazilian coast): epidemiologic, clinic and therapeutic aspects of 23 stings in humans. AB - Fishes of the family Scorpaenidae are responsible for severe injuries and occasionally deaths in humans around the world. The more venomous fishes on the Brazilian coast and in the Southwestern Atlantic region are classified in the genus Scorpaena (family Scorpaenidae). However, there are few studies on the venomous apparatus, the effects of the venom, or clinical aspects of human envenoming provoked by Atlantic scorpionfishes. In this communication, the authors present 23 accidents caused by scorpionfishes of the genus Scorpaena among fishermen, and report the species that provoked the injuries, the circumstances of contacts, the clinical aspects observed and the therapeutic measures utilized for control of the symptoms of the victims. The intense pain and the systemic findings observed in the patients were very frequent and we think that the injuries provoked by scorpionfishes should be considered the most important manifestations caused by venomous fishes of the East Atlantic Ocean. PMID- 12893065 TI - Responses of antioxidant systems in the hepatocytes of common carp (Cyprinus carpio L.) to the toxicity of microcystin-LR. AB - The freshwater, bloom-forming cyanobacterium (blue-green alga) Microcystis aeruginosa produces a peptide hepatotoxin, which causes the damage of animal liver. Recently, toxic Microcystis blooms frequently occur in the eutrophic Dianchi Lake (300 km2 and located in the South-Western of China). Microcystin-LR from Microcystis in Dianchi was isolated and purified by high performance liquid chromatography (HPLC) and its toxicity to mouse and fish liver was studied (Li et al., 2001). In this study, six biochemical parameters (reactive oxygen species, glutathione, superoxide dismutase, catalase, glutathione peroxide and glutathione S-transferase) were determined in common carp hepatocytes when the cells were exposed to 10 microg microcystin-LR per litre. The results showed that reactive oxygen species (ROS) contents increased by more than one-time compared with the control after 6 h exposure to the toxin. In contrast, glutathione (GSH) levels in the hepatocytes exposed to microcystin-LR decreased by 47% compared with the control. The activities of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxide (GSH-Px) increased significantly after 6 h exposure to microcystin-LR, but glutathione S-transferase (GST) activity showed no difference from the control. These results suggested that the toxicity of microcystin-LR caused the increase of ROS contents and the depletion of GSH in hepatocytes exposed to the toxin and these changes led to oxidant shock in hepatocytes. Increases of SOD, CAT and GSH-Px activities revealed that these three kinds of antioxidant enzymes might play important roles in eliminating the excessive ROS. This paper also examined the possible toxicity mechanism of microcystin-LR on the fish hepatocytes and the results were similar to those with mouse hepatocytes. PMID- 12893066 TI - Implication of brevetoxin B1 and PbTx-3 in neurotoxic shellfish poisoning in New Zealand by isolation and quantitative determination with liquid chromatography tandem mass spectrometry. AB - Brevetoxin B1 (BTX-B1) was isolated from Austrovenus stutchburyi following the 1992-1993 outbreak of neurotoxic shellfish poisoning (NSP) in New Zealand. We report here the first isolation of PbTx-3 from the same shellfish and the development of a procedure for quantitative determination of PbTx-3 and BTX-B1. PbTx-3 was isolated by chromatography on columns of SiO2, ODS, and LH-20, followed by reverse-phase HPLCs. In mass spectrometry (MS) with an electrospray ionization (ESI) interface operating in the positive or negative ion mode, the abundant protonated ion [M+H]+ of PbTx-3 (m/z 897) and the de-sodiated ion [M-Na] of BTX-B1 (m/z 1016) were generated, respectively. These served as precursor ions for collision-induced dissociation, and the product ions of m/z 725 from PbTx-3 and m/z 80 from BTX-B1 were identified, allowing unambiguous confirmation of these toxins by selected reaction monitoring liquid chromatography-tandem mass spectrometry (SRM LC-MS/MS) analysis. The determination limits were 0.4 and 2 ng/g for BTX-B1 and PbTx-3 at a signal-to-noise ratio of five, respectively. This LC-MS/MS method was successfully applied to determine BTX-B1 and PbTx-3 in the NSP-associated toxic shellfish. BTX-B1 was found in both A. stutchburyi and Perna canaliculus, but not in Crassostrea gigas, while PbTx-3 was found in all three. PMID- 12893068 TI - Bibliography of toxinology. PMID- 12893067 TI - The first identification of azaspiracids in shellfish from France and Spain. AB - Incidents of human intoxications throughout Europe, following the consumption of mussels have been attributed to Azaspiracid Poisoning (AZP). Although first discovered in Ireland, the search for the causative toxins, named azaspiracids, in other European countries has now led to the first discovery of these toxins in shellfish from France and Spain. Separation of the toxins, azaspiracid (AZA1) and analogues, AZA2 and AZA3, was achieved using isocratic reversed-phase liquid chromatography coupled, via an electrospray ionisation source, to an ion-trap mass spectrometer. Azaspiracids were identified in mussels (Mytilus galloprovincialis), 0.24 microg/g, from Galicia, Spain, and scallops (Pecten maximus), 0.32 microg/g, from Brittany, France. Toxin profiles were similar to those found in the equivalent shellfish in Ireland in which AZA1 was the predominant toxin. PMID- 12893070 TI - Immunohistochemical analysis of cyclooxygenase-2 expression in premalignant and malignant esophageal glandular and squamous lesions in Cixian, China. AB - Increased cyclooxygenase-2 (COX-2) expression has been observed in both squamous cell carcinoma (SCC) and adenocarcinoma (AC) in Western countries, and COX-2 inhibitors have been considered as potential chemopreventive agents for esophageal cancers. Since chemoprevention often targets the premalignant lesions in high-risk population, it is worthwhile to study COX-2 expression in a spectrum of premalignant and malignant lesions obtained from the high-risk populations. In this study, biopsy samples were taken from 240 subjects identified by screening of the high-risk population in Cixian, China, including 27 normal, 29 with squamous hyperplasia, 84 with dysplasia (31 low grade and 53 high grade), 30 with carcinoma in situ, and 70 with invasive carcinoma (60 SCC and 10 AC). For comparison, tissue samples were also collected from He Lon Jiang Province, a low risk population in China, including 10 patients with invasive SCC, 20 patients with AC, and 17 patients with Barrett's esophagus. The COX-2 protein expression was examined by immunohistochemistry. Using 10% staining as a threshold, 9 of 10 (90%) invasive SCC from low-risk population were COX-2 positive. However, no positive COX-2 staining was seen on normal, hyperplastic, dysplastic, and in situ squamous lesions from the high-risk population, and only 4 of 60 (6%) invasive SCC exhibited positive COX-2 staining. For glandular lesions, 6 of 10 (60%) AC from high-risk area and 15 of 20 (75%) from low-risk area showed positive COX-2 staining, and 12 of 17 (70%) premalignant Barrett's esophagus were also positive. Our findings show that COX-2 expression various in squamous lesions from high- and low-risk areas, but not in glandular lesions. Additional studies are needed to fully explore the mechanisms that are associated with the different COX-2 immunohistochemical staining patterns in esophageal squamous lesions from low- and high-risk populations. PMID- 12893072 TI - Atypical Carney's triad with coincidental clear cell renal carcinoma in an 84 year old patient-a finding at autopsy. AB - Carney's triad is a syndrome of unknown etiology, representing a combination of gastrointestinal stromal tumors, bronchial chondromas and vagal, adrenal or paraadrenal paragangliomas. Two of the Carney's triad components-the paragangliomas and the gastrointestinal stromal tumors-are potentially lethal. Since its first description in 1977, 79 cases have been reported so far. We report an 84-year-old male patient, who died of a hypertensive cerebral hemorrhage. Well-differentiated clear cell carcinoma of the right kidney, chondroma of the right bronchus and multiple jejunal stromal tumors were detected at autopsy. To our knowledge, this is the first report of a coincidental clear cell renal carcinoma in a patient with an atypical Carney's triad. PMID- 12893071 TI - Similar BCL-X but different BCL-2 levels in the two age groups of north African nasopharyngeal carcinomas. AB - Nasopharyngeal carcinomas (NPCs) are consistently associated with the Epstein Barr virus (EBV). As Bcl-2 and Bcl-X are co-expressed in EBV-transformed B lymphocytes, we attempted to determine their status in malignant NPC cells. A retrospective series of 100 NPC specimens from untreated Tunisian patients was investigated by immuno-histochemistry. Twenty seven of the patients were below 30 years old and therefore classified in the "juvenile" form of north African NPCs. Bcl-2 and Bcl-X expression was assessed semi-quantitatively using a score based on the percentage of positive cells and staining intensity. Intense Bcl-X expression was detected in malignant cells of 100% biopsy samples with similar scores for patients below 30 years or those aged 30 or over. Bcl-2 was detected in 89% biopsies but its expression differed considerably between the samples. The average Bcl-2 score was much lower for patients under 30 years (4.4+/-1.5 compared to 6.5+/-2 for older patients; P<10(-6)). Multivariate analysis demonstrated that no other clinical parameter, except the primary tumor size, was correlated to the Bcl-2 score. Bcl-X and Bcl-2 are co-expressed in 89% of NPCs whereas their expression is mutually exclusive in other head and neck carcinomas (particularly squamous cell carcinomas, SCC). The constantly high expression of Bcl-X is consistent with it being induced by the EBV protein Epstein-Barr nuclear antigen 1 (EBNA1), as recently reported in a murine model. The contrasted levels of Bcl-2 expression in the two age groups strengthen the hypothesis that these clinical forms result from distinct oncogenic mechanisms. PMID- 12893073 TI - Prevalence of proximal adenomas after an adenoma is found on flexible sigmoidoscopy. AB - BACKGROUND: Adenomatous polyps are a precursor of colorectal cancer and a frequent finding on screening flexible sigmoidoscopy (FS). Performance of colonoscopy when a diminutive (<6mm) adenoma is found on FS has been the subject of considerable debate. METHODS: We retrospectively reviewed the data from our colorectal cancer screening program for patients with adenoma(s) found on FS. Patients were divided into three groups based on FS findings: (1) an adenoma <6mm in size, (2) multiple non-advanced adenomas or an adenoma 6-10mm in size, or (3) advanced adenoma defined as an adenoma >10mm or with villous histology or high grade dysplasia or cancer. A comparison of the proximal findings was then made. RESULTS: 5291 FS reports were reviewed with 606 (12%) patients having at least one adenoma. Colonoscopy reports were available in 550 patients. Of the 258 patients with a diminutive distal adenoma, 69 (27%) had a proximal adenoma and 13 (5%) had an advanced proximal adenoma on colonoscopy. Of the 164 patients with an adenoma 6-10mm or multiple non-advanced adenomas, 59 (36%) had a proximal adenoma and 13 (8%) had an advanced proximal adenoma. Of the 128 patients with a distal advanced adenoma, 58 (45%) had a proximal adenoma and 15 (12%) had an advanced proximal adenoma. The increase in proximal adenomas across the three groups was significant (P=0.001), and there was a trend for increased prevalence of advanced adenomas (P=0.061). CONCLUSIONS: The prevalence of proximal adenomas increased significantly with more advanced lesions found distally at FS, and there was a trend towards a higher prevalence of advanced proximal adenomas. Based on current guidelines, flexible sigmoidoscopy is a screening option that can be used to identify average-risk patients at increased risk of proximal neoplasia. PMID- 12893074 TI - Secular stability and reliability of measurements of the percentage of dense tissue on mammograms. AB - Elevated mammographic density is associated with increased risk of breast cancer. We conducted a reliability study on mammographic density assessments to determine their potential usefulness for projecting individual breast cancer risk. We used baseline screening mammograms from 7251 women in the Breast Cancer Detection Demonstration Project (BCDDP). Repeated measurements from the same images were used to assess measurement variability by an experienced evaluator. Intraclass correlations of assessments over time usually exceeded 0.9, indicating usefulness for prospective applications. Data also indicated it may be reasonable to include cases identified in the first year of screening together with other cases in developing a risk model. Older ages and increased weight were associated with decreased mammographic density. The density of the right breast slightly exceeded that of the left. Among women who developed breast cancer, the baseline mammographic density of the ipsilateral (diseased) breast was 0.53 (95% confidence interval (CI) 0.20-0.86) percentage units higher than in the contralateral breast. PMID- 12893075 TI - The role of various biomarkers in the evaluation of styrene genotoxicity. AB - We evaluated our data on the occupational exposure to styrene in lamination workers. The battery of parameters included markers of external and internal exposure and biomarkers of biological effects and susceptibility. DNA repair capacities have been determined in both exposed and control groups. Styrene workplace concentration significantly correlated with styrene concentration in blood, exhaled air and urinary mandelic acid. Haemoglobin and O(6)-styrene oxide (SO)-guanine DNA adducts were significantly higher in exposed subjects as compared to controls and correlated with exposure parameters. In styrene-exposed workers 1-SO-adenine DNA adducts were detected (2.6 per 10(9) dNp), while in controls these adducts were below the detection limit. 1-SO-adenine adduct levels were affected by both acute and cumulative exposure (P=0.001, F=86.0 and P=0.017, F=59.0, respectively) and associated with cytochrome P450 2E1 (CYP2E1) polymorphisms (R(2)=0.442). Mutant frequencies (MF) at the hypoxanthine-guanine phosphoribosyltransferase (HPRT) locus appeared to accumulate with exposure over time and were associated with glutathione S-transferase P1 (GSTP1) polymorphism. DNA repair capacity increased with the exposure, except for the group exposed to the highest styrene concentration. In this particular group, increased DNA repair capacity to remove oxidative DNA damage was found. PMID- 12893076 TI - Antibodies to HSP70 and HSP90 in serum in non-small cell lung cancer patients. AB - Heat shock proteins (HSPs) are components of a physiologic stress response that are also over-expressed in various cancers including non-small cell lung cancer (NSCLC). During NSCLC serum-antibody screening of a NSCLC cDNA T7 phage library for immunogenic proteins we isolated HSP70 and HSP90 proteins. Isolation of these proteins suggested that corresponding antibodies could be elevated in NSCLC patient sera, a novel finding that could pilot their use as markers of NSCLC. We showed histochemically that patient sera were more reactive with each phage expressed protein than normal sera. Antibody affinity for each phage-expressed protein was confirmed by limiting the dilution of individual sera assayed by Ab enzyme-linked immunosorbent assay (ELISA). Sera from 49 NSCLC patients assayed by Ab ELISA and normalized to 40 controls showed that HSP70 antibodies were significantly greater in patient sera than in normals (P=0.0002), while HSP90 antibodies were not significantly different (P=0.11). Analysis of the results with logistic regression and receiver operating characteristics (ROC) curves showed that HSP70 antibodies were modest markers of NSCLC (sensitivity 0.74 and specificity 0.73; area under the curve or AUC=0.731), while HSP90 antibodies appeared to be poor in both criteria with an AUC of 0.602. Further evaluation of HSP70 antibodies as potential markers of disease may be rational. PMID- 12893077 TI - Microphysiological testing for chemosensitivity of living tumor cells with multiparametric microsensor chips. AB - A constraint in the reliability of predictive chemosensitivity assays is linked to the fact that they analyze only a single cellular or biochemical parameter. A multiparametric test system using microsensor chips has been developed which can detect online microphysiological changes in living cells. Tumor cells were grown directly on glass- or silicon-based electronic sensor chips. Changes in extracellular pH and pO(2), reflecting metabolic activities, and changes in impedance, reflecting morphological properties, were monitored. In this study, colon and breast cancer cells as well as doxorubicin-sensitive and doxorubicin resistant sarcoma cell lines were exposed to cytochalasin B, chloroacetaldehyde, or doxorubicin. Results show (1) reduction in medium acidification, (2) marked and rapid changes in O(2) consumption, and (3) modulations in impedance correlating with morphological changes observed in the microscope. Drug-resistant cells do not show these changes. Therefore, this microphysiological monitoring is a versatile tool for chemosensitivity testing of tumor cells. PMID- 12893079 TI - Influence of parotid-sparing radiotherapy on xerostomia in head and neck cancer patients. AB - Radiotherapy (RT) for head and neck cancers causes permanent salivary gland dysfunction (SGD) and xerostomia. We have previously demonstrated the effectiveness of parotid-sparing RT on salivary function. The aim of this was to characterize the relationship between radiation dosages to parotid glands, SGD, xerostomia, and impaired quality of life (QOL). Ninety-three patients received unilateral (n=38) and bilateral (n=44) neck RT with parotid-sparing techniques, or standard three-field technique RT (n=11). Unstimulated and stimulated parotid saliva was collected pre-RT and 1 year post-RT. Assessment of QOL and xerostomia was conducted with three questionnaires. The results demonstrated that reduced radiation dosages to parotid glands were strongly associated with percentage of baseline parotid flow rates measured at 1 year post-RT. Unilateral and bilateral neck RT with parotid-sparing techniques were successful in preserving salivary output, compared to standard three beam RT techniques. Lower radiation dose to contralateral parotid glands was associated with greater percentage of baseline salivary flow rates at 1 year post-RT, fewer xerostomic complaints, and an enhanced QOL. PMID- 12893078 TI - Folic acid mediated attenuation of loss of heterozygosity of DCC tumor suppressor gene in the colonic mucosa of patients with colorectal adenomas. AB - Loss of heterozygosity (LOH) and/or inactivation of tumor suppressor genes are implicated in the initiation and progression of many malignancies, including colorectal cancer. Although accumulating evidence suggests a chemopreventive role for folate in colorectal cancer, regulatory mechanisms are poorly understood. The primary objective of the current investigation was to determine whether folic acid would prevent LOH of the three tumor suppressor genes, deleted in colorectal cancer (DCC), adenomatous polyposis coli (APC) and p53 in macroscopically normal appearing rectal mucosa of patients with adenomatous polyps. In addition, the effect of folic acid on rectal mucosal proliferation was determined. Twenty patients were randomized in a double-blind study to receive either folic acid 5mg once daily or identical placebo tablets for 1 year. Genomic DNA and total protein were extracted from the rectal mucosa at baseline and after 1 year of treatment and analyzed for LOH and protein levels of APC, DCC and p53 genes. In addition, paraffin-embedded mucosal specimens were analyzed for proliferating cell nuclear antigen (PCNA) immunoreactivity, as a measure of cellular proliferative activity. Folate supplementation prevented LOH of DCC gene in five out of five (100%) patients who demonstrated baseline heterozygosity, whereas two out of four (50%) placebo-treated patients with baseline heterozygosity demonstrated allelic loss. Mucosal protein levels of DCC were also reduced in 7 of 10 (70%) placebo-treated patients compared to only 2 of 10 (20%) of patients treated with folate. Levels increased, however, in eight and three patients in the folic acid and placebo groups, respectively (P<0.02). Folic acid caused no change in allelic status of either APC or p53 gene. Folate supplementation caused a small, but not statistically significant, 16% reduction in mucosal proliferation, whereas placebo treatment resulted in a 88% (P<0.05) increase in this parameter, when compared with the corresponding baseline values. Our results indicate that folic acid prevents an increase in proliferation and arrests LOH of DCC gene and also stabilizes its protein in normal appearing rectal mucosa of patients with colorectal adenomas. Taken together, our data suggest that one of the ways folate may exert its chemopreventive effect is by stabilizing certain tumor suppressor gene(s) and preventing further increases in proliferation. PMID- 12893081 TI - Impact factors-and all that jazz. PMID- 12893080 TI - Development and reliability of a brief skin cancer risk assessment tool. AB - This study aimed to develop and pilot test a brief skin cancer risk assessment tool (BRAT), a self-administered instrument that can be reliably used to assess skin cancer risk. To develop the BRAT, we critically reviewed published literature on risk factors; formulated a draft questionnaire; pilot tested the questionnaire; and retested 1 month later. The BRAT items address the key risk factors for melanoma and other keratinocyte skin cancers: ethnicity, personal and family history of skin cancer, mole count, freckles, childhood residence, sunburn history, and sun sensitivity factors (skin color, natural hair color, ease of sunburning and tanning). One hundred sixty-five persons completed the initial BRAT pilot study, and 52 additional people at moderate- or high-risk completed a second BRAT pilot study. Results were as follows: using a dichotomous risk measure, about 90% of subjects would be correctly classified at baseline and follow-up. Weighted kappa for the total BRAT score (0.41-0.68) and for individual items (0.57-0.99) were fair to good, as were correlation coefficients. The BRAT has acceptable to good reproducibility. Reliability statistics compared favorably with those reported in the literature for similar measures. PMID- 12893082 TI - Structural characterization of the Fpg family of DNA glycosylases. AB - Until recently, the Fpg family was the only major group of DNA glycosylases for which no structural data existed. Prototypical members of this family, found in eukaryotes as well as prokaryotes, have now been crystallized as free proteins and as complexes with DNA. In this review, we analyze the available structural information for formamidopyrimidine-DNA glycosylase (Fpg) and endonuclease VIII (Nei). Special emphasis is placed on mechanisms by which these enzymes recognize and selectively excise cognate lesions from oxidatively damaged DNA. The problem of lesion recognition is considered in two parts: how the enzyme efficiently locates a single lesion embedded in a vast excess of DNA; and how the lesion is accommodated in a pocket near the active site of the enzyme. Although all crystal structures reported to date for the Fpg family lack the damaged base, functionally important residues that participate in DNA binding and enzyme catalysis have been clearly identified and other residues, responsible for substrate specificity, have been inferred. PMID- 12893083 TI - Initiation of repair of A/G mismatches is modulated by sequence context. AB - The efficiency of DNA glycosylases to initiate base excision repair (BER) has been demonstrated to be modulated by the precise sequence context in which the lesion or mismatch is located. In the case of DNA containing an A/G mismatch, in which the recognition and excision of adenine from the mismatch is mediated by the Escherichia coli MutY enzyme, not only does the local sequence context affect the strength of base stacking interactions, but it also modulates the syn/anti conformation around the glycosyl bond of the bases in the mispair. Utilizing prior NMR data to identify DNA sequence contexts that adopt either an anti/anti or a syn/anti configuration at an A/G mismatch, we tested the hypothesis that the initial equilibrium of the mismatched base orientations would modulate the overall efficiency of glycosyl bond scission. By systematically varying the sequence context around a central A/G mismatch within a 30-mer duplex DNA, significant kinetic differences were observed that were consistent with this hypothesis. Since the relative efficiency of the kinetics fell into only two groupings, a NMR study was conducted on a DNA sequence context of unknown syn/anti conformation. These data established that the relative syn/anti conformation did not correlate with the excision efficiency, as well as there being a lack of correlation between kinetics and thermal stability of these DNAs. PMID- 12893084 TI - Modification of the alkaline Comet assay to allow simultaneous evaluation of mitomycin C-induced DNA cross-link damage and repair of specific DNA sequences in RT4 cells. AB - The alkaline Comet assay is a simple, sensitive method for measuring the extent of DNA strand breaks in individual cells. Several modifications to the original assay have been developed to increase its applications. One such modification allows the measurement of DNA cross-links by assessing the relative reduction in DNA migration induced by a strand-breaking agent. Another modification includes the application of fluorescent in situ hybridisation (FISH) to investigate the localisation of specific gene domains within a cell. Although several studies have used these approaches separately, no report to date has combined these two versions of the Comet assay. The current study describes the modification of the Comet assay, to allow both measurement of mitomycin C (MMC)-induced cross-links and the subsequent application of FISH to study repair in the TP53 gene region. RT4 human bladder cancer cells were treated with 0, 5, 50 and 200 microg/ml MMC to study dose response, whilst for cross-link repair studies, they were treated with 50 microg/ml MMC and allowed to repair for up to 24 h. A clear dose response to MMC was displayed, demonstrable by a marked reduction in DNA migration, whilst repair studies showed that MMC-induced cross-links take at least 24 h to repair fully in RT4 cells. For Comet-FISH experiments, the number and location of TP53 hybridisation spots was also recorded for each cell. In dose response experiments, the number of spots per cell, and per Comet tail, decreased as MMC dose increased. In repair experiments, the number of spots, particularly in the Comet tail, increased as repair time increased. Furthermore, our results suggest that repair of the TP53 gene region is most rapid within the first 4 h following MMC treatment. We conclude that the novel experimental protocol presented here has considerable potential in evaluating DNA damage and sequence-related repair responses to cross-linking agents. PMID- 12893085 TI - Inhibition of p53, p21 and Bax by pifithrin-alpha does not affect UV induced apoptotic response in CS-B cells. AB - Cockayne syndrome (CS) is a human autosomal recessive disorder characterized by many neurological and developmental abnormalities. CS cells are defective in transcription coupled repair (TCR) pathway that removes DNA damage from the transcribed strand of active genes. In spite of a TCR deficiency at the cellular level, CS patients do not develop cancer. The lack of cancer incidence in CS patients may be due to the selective elimination of cells by an apoptotic pathway. In order to verify the role of p53-associated pathway in ultraviolet (UV) induced apoptosis in human CS-B cells, the expression of p53 and p53 responsive genes was analysed in UV irradiated human cells after treatment with pifithrin-alpha (PFTalpha). PTFalpha effectively inhibited the induction of p53, p21 and Bax after UV treatment without affecting the apoptotic response in CS-B cells. Our results indicate that the p53-associated pathway involving p21 and Bax does not largely contribute to UV induced apoptosis in TCR defective human CS-B cells. PMID- 12893086 TI - From genotype to phenotype: correlating XRCC1 polymorphisms with mutagen sensitivity. AB - This study correlated the extent of induced in vitro chromosomal damage, assessed by the mutagen sensitivity assay, with genotypes of the X-ray repair cross complementing group 1 (XRCC1) gene, which encodes for a base excision repair protein. There are two common polymorphisms that cause amino acid substitutions in XRCC1, one at codon 194 in exon 6 and another at codon 399 in exon 10. We genotyped these two polymorphisms in 524 healthy subjects and performed mutagen sensitivity assays using both bleomycin and benzo[a]pyrene-diol-epoxide (BPDE) as challenge mutagens. Our results showed that individuals with the wildtype exon 6 Arg/Arg exhibited significantly higher values of chromosomal breaks per cell (b/c) than those with one or two variant Trp alleles (P=0.005 for bleomycin and P=0.05 for BPDE). For the exon 10 polymorphism, subjects who were Gln/Gln homozygotes had higher b/c than did those with other genotypes, with evidence of a gene dosage effect. When we combined the two polymorphic sites and used the exon 6 Arg/Trp and Trp/Trp and exon 10 Arg/Arg genotypes as the reference category, these differences were enhanced for bleomycin sensitivity (P for trend = 0.032), but not for BPDE sensitivity (P for trend = 0.821). These data are biologically plausible since codon 399 is located within the BRCA1 C-terminus functional domain and codon 194 is in the linker region of the XRCC1 N-terminal functional domain. To our knowledge, this is the largest study conducted evaluating the functional relevance of these polymorphisms. PMID- 12893087 TI - Translesion replication in cisplatin-treated xeroderma pigmentosum variant cells is also caffeine-sensitive: features of the error-prone DNA polymerase(s) involved in UV-mutagenesis. AB - Patients with xeroderma pigmentosum variant (XP-V) have a higher risk to skin cancer and XP-V cells are extremely mutable by ultraviolet (UV). The defective gene encodes a DNA polymerase (Poleta) which catalyzed relatively accurate translesion synthesis past the cyclobutane dimer of UV-lesions instead of the replicative polymerase(s) that stalled just before the lesion. Pulse-chase studies have shown that translesion replication in XP-V cells is delayed, but does not completely cease. Taking these results together, error-prone polymerase(s) are plausively involved in the UV-mutagenesis in XP-V devoid of Poleta. However, less is known about the polymerase(s) in vivo. Using an alkaline sucrose density gradient centrifugation (ASDG) technique, translesion replication is detected in the two XP-V strains XP30RO and XP115LO. As reported by Lehmann et al. [Proc. Natl. Acad. Sci. U.S.A. 72 (1975): 219] in XP-V; (i) smaller replication products were accumulated after UV irradiation; (ii) the elongation of these products was delayed; (iii) the elongation was markedly inhibited by caffeine. XP-V cells UV-irradiated at mid-S phase were normally S-arrested, and no "override" by caffeine (i.e. abrogation of the S-checkpoint) was observed by flow cytometry, suggesting that caffeine does not act via cdc kinase here; (iv) butylphenyldeoxyguanosine (BuPGdR) inhibited elongation of replication products only in UV-irradiated XP-V cells; (v) dideoxycytidine or dideoxyinosine had no effect on this process in either normal or XP-V cells. Next, similar phenomena to UV (all of above i to v) were observed also in cisplatin-treated XP-V cells. Pol eta was indicated to participate in cisplatin-induced translesion replication in normal cells. Summing up the above results, the polymerase(s) which work in translesion replication in XP-V are probably BuPGdR-sensitive, insensitive to dideoxynucleotides and can bypass also cisplatin-lesions. To date, several polymerases capable of lesion-bypass synthesis have been isolated. The features presented here are quite useful for identifying the error-prone polymerase(s) involved in UV-mutagenesis. PMID- 12893088 TI - Complementary functions of the Saccharomyces cerevisiae Rad2 family nucleases in Okazaki fragment maturation, mutation avoidance, and chromosome stability. AB - Rad2 family nucleases, identified by sequence similarity within their catalytic domains, function in multiple pathways of DNA metabolism. Three members of the Saccharomyces cerevisiae Rad2 family, Rad2, Rad27, and exonuclease 1 (Exo1), exhibit both 5' exonuclease and flap endonuclease activities. Deletion of RAD27 results in defective Okazaki fragment maturation, DNA repair, and subsequent defects in mutation avoidance and chromosomal stability. However, strains lacking Rad27 are viable. The expression profile of EXO1 during the cell cycle is similar to that of RAD27 and other genes encoding proteins that function in DNA replication and repair, suggesting Exo1 may function as a back up nuclease for Rad27 in DNA replication. We show that overexpression of EXO1 suppresses multiple rad27 null mutation-associated phenotypes derived from DNA replication defects, including temperature sensitivity, Okazaki fragment accumulation, the rate of minichromosome loss, and an elevated mutation frequency. While generally similar findings were observed with RAD2, overexpression of RAD2, but not EXO1, suppressed the MMS sensitivity of the rad27 null mutant cells. This suggests that Rad2 can uniquely complement Rad27 in base excision repair (BER). Furthermore, Rad2 and Exo1 complemented the mutator phenotypes and cell cycle defects of rad27 mutant strains to differing extents, suggesting distinct in vivo nucleic acid substrates. PMID- 12893090 TI - Mood disorders and medical illness: a major public health problem. PMID- 12893091 TI - Recognizing and meeting the needs of patients with mood disorders and comorbid medical illness: a consensus conference of the Depression and Bipolar Support Alliance. PMID- 12893092 TI - Depression and comorbid medical illness: the National Institute of Mental Health perspective. PMID- 12893093 TI - Depression in cardiovascular disease: can the risk be reduced? PMID- 12893094 TI - Mood disorders and cancer: a National Cancer Institute perspective. PMID- 12893095 TI - Comorbid mood disorders and medical illness: a Food and Drug Administration perspective. PMID- 12893096 TI - Mood disorders and allostatic load. AB - The brain controls both the physiologic and the behavioral coping responses to daily events as well as major stressors, and the nervous system is itself a target of the mediators of those responses through circulating hormones. The amygdala and hippocampus interpret what is stressful and regulate appropriate responses. The amygdala becomes hyperactive in posttraumatic stress disorder (PTSD) and depressive illness, and hypertrophy of amygdala nerve cells is reported after repeated stress in an animal model. The hippocampus expresses adrenal steroid receptors. It undergoes atrophy in several psychiatric disorders and responds to repeated stressors with decreased dendritic branching and reduction in number of neurons in the dentate gyrus. Stress promotes adaptation ("allostasis"), but a perturbed diurnal rhythm or failed shutoff of mediators after stress ("allostatic state") leads, over time, to wear and tear on the body ("allostatic load"). Neural changes mirror the pattern seen in the cardiovascular, metabolic, and immune systems, that is, short-term adaptation versus long-term damage. Allostatic load leads to impaired immunity, atherosclerosis, obesity, bone demineralization, and atrophy of nerve cells in brain. Allostatic load is seen in major depressive illness and may also be expressed in other chronic anxiety disorders such as PTSD and should be documented. PMID- 12893097 TI - Social and economic burden of mood disorders. AB - Social and economic effects of mood disorders include functional impairment, disability or lost work productivity, and increased use of health services. Evidence for these impacts includes cross-sectional studies, longitudinal studies, and true experiments (randomized trials of specific treatments or treatment programs). With respect to unipolar depression, strong evidence demonstrates that depression is associated with significant functional impairment and that effective treatment helps to restore function. Studies of the effect of depression on work disability and health care costs show strong cross-sectional associations (i.e., greater disability and higher costs among those with depression) and longitudinal associations (i.e., improvement in depression is associated with reduced disability and lower costs). All of these findings regarding unipolar depression seem as consistent in the subgroup of patients with comorbid chronic medical illness as in the total population with depressive disorders. Fewer data are available regarding social and economic burden of bipolar disorder, but available data show cross-sectional associations between mood symptoms and functional impairment, disability, and health care costs. Taken together, these data describe the substantial social and economic burden of mood disorders and the potential benefits of more effective treatment. We must recall, however, that economic benefits of treatment for mood disorders are secondary to the principal objective of relieving human suffering. PMID- 12893098 TI - Clinical and health services relationships between major depression, depressive symptoms, and general medical illness. AB - Patients with chronic medical illness have a high prevalence of major depressive illness. Major depression may decrease the ability to habituate to the aversive symptoms of chronic medical illness, such as pain. The progressive decrements in function associated with many chronic medical illnesses may cause depression, and depression is associated with additive functional impairment. Depression is also associated with an approximately 50% increase in medical costs of chronic medical illness, even after controlling for severity of physical illness. Increasing evidence suggests that both depressive symptoms and major depression may be associated with increased morbidity and mortality from such illnesses as diabetes and heart disease. The adverse effect of major depression on health habits, such as smoking, diet, over-eating, and sedentary lifestyle, its maladaptive effect on adherence to medical regimens, as well as direct adverse physiologic effects (i.e., decreased heart rate variability, increased adhesiveness of platelets) may explain this association with increased morbidity and mortality. PMID- 12893099 TI - Epidemiology of comorbid coronary artery disease and depression. AB - This article reviews the epidemiology of comorbid coronary artery disease and unipolar depression. Both major depression and subsyndromal depressive symptoms will be considered; unless otherwise specified, the term depression will be used to designate all depressive states, including major depressive disorder, minor depression, dysthymia, and other subsyndromal forms of depression. While 17% to 27% of patients with coronary artery disease have major depression, a significantly larger percentage has subsyndromal symptoms of depression. Patients with coronary artery disease and depression have a twofold to threefold increased risk of future cardiac events compared to patients without depression, independent of baseline cardiac dysfunction. The relative risk for the development of coronary artery disease conferred by depression in patients initially free of clinical cardiac disease is approximately 1.5, independent of other known risk factors for coronary disease. In the discussion, special attention will be paid to the interactions of both gender and age with depression and coronary artery disease risk. Scrutiny of the role of confounding risk factors is presented, such as global burden of comorbid medical illness and modification of traditional risk factors, which may, in part, mediate the effect of depression on coronary artery disease. PMID- 12893100 TI - Depression, mortality, and medical morbidity in patients with coronary heart disease. AB - There is substantial evidence that depression is a risk factor for cardiac morbidity and mortality, both for patients without clinical evidence of coronary heart disease at index examination and for patients with established coronary disease. The relationship is most apparent for patients with a recent acute myocardial infarction. Many questions about the impact of depression on heart disease remain unresolved. PMID- 12893101 TI - Depression and cardiovascular disease: mechanisms of interaction. AB - This article explores the relationship between depression and cardiovascular disease from a mechanistic standpoint. Depression and cardiovascular disease are two of the most prevalent health problems in the United States and are the two leading causes of disability both in the United States and worldwide. Although depression is a known risk factor for the development of cardiovascular disease, as well as an independent predictor of poor prognosis following a cardiac event, the mechanistic relationship between the two remains unclear. Depression is associated with changes in an individual's health status that may influence the development and course of cardiovascular disease, including noncompliance with medical recommendations, as well as the presence of cardiovascular risk factors such as smoking and hypertension. In addition, depression is associated with physiologic changes, including nervous system activation, cardiac rhythm disturbances, systemic and localized inflammation, and hypercoagulability, that negatively influence the cardiovascular system. Further, stress may be an underlying trigger that leads to the development of both depression and cardiovascular disease. This article reviews seven potential mechanisms for the relationship between depression and cardiovascular disease and presents the available evidence surrounding each mechanism. Finally, future directions for research are discussed. PMID- 12893102 TI - Treatment of depression in patients with heart disease. AB - There is a growing body of evidence that depression significantly and adversely affects cardiovascular health. Perhaps the most prominent finding is the documented increase in mortality rate in patients with depression after myocardial infarction. The critical questions of interest to both the clinician and researcher are whether there are safe and effective treatments for depression in patients with heart disease and whether treatment of depression reduces the increased risk of cardiac morbidity and mortality. Though the data are limited and are primarily from open or comparator trials, the tricyclic antidepressants (TCAs), selective serotonin reuptake inhibitors (SSRIs), and specific psychotherapies appear to be effective for treatment of depression in patients with ischemic heart disease (IHD), and response rates are comparable to those reported in depressed patients without heart disease; however, there has been only one placebo-controlled trial to date, and therefore it is premature to come to definitive conclusions regarding the efficacy of antidepressant therapies in this patient population. With respect to safety, the TCAs are associated with documented adverse cardiovascular effects, including increases in heart rate, orthostatic hypotension, and conduction delays. Use of TCAs in patients with IHD carries a proven increased risk of cardiac morbidity and perhaps of mortality as well. The SSRIs appear to be relatively safe and effective treatment for depression in patients with comorbid IHD. PMID- 12893103 TI - Depression and cancer: mechanisms and disease progression. AB - Depression and cancer commonly co-occur. The prevalence of depression among cancer patients increases with disease severity and symptoms such as pain and fatigue. The literature on depression as a predictor of cancer incidence is mixed, although chronic and severe depression may be associated with elevated cancer risk. There is divided but stronger evidence that depression predicts cancer progression and mortality, although disentangling the deleterious effects of disease progression on mood complicates this research, as does the fact that some symptoms of cancer and its treatment mimic depression. There is evidence that providing psychosocial support reduces depression, anxiety, and pain, and may increase survival time with cancer, although studies in this latter area are also divided. Psychophysiological mechanisms linking depression and cancer progression include dysregulation of the hypothalamic-pituitary-adrenal axis, especially diurnal variation in cortisol and melatonin. Depression also affects components of immune function that may affect cancer surveillance. Thus, there is evidence of a bidirectional relationship between cancer and depression, offering new opportunities for therapeutic intervention. PMID- 12893104 TI - Depression in cancer: new developments regarding diagnosis and treatment. AB - Considerable data demonstrate the high prevalence of symptoms of depression in patients with a wide variety of neoplastic disorders. Moreover, the dire consequences of these depressive symptoms in cancer patients have been well documented. Recent conceptual developments in the potential contributing mechanisms include increasing appreciation of the possibility that behavioral alterations in cancer patients may represent a "sickness syndrome" that results from activation of the inflammatory cytokine network. This sickness syndrome, which has been well documented in patients and laboratory animals exposed to inflammatory cytokines, includes symptoms that overlap with those seen in major depression. Conceptualizing these symptoms as components of cytokine-mediated sickness behavior has several important, and potentially novel, implications, including 1) an expansion of the neurobehavioral symptoms that are relevant to diagnosis and treatment; and 2) an increased appreciation of the potential diagnostic utility of peripheral markers of inflammation, as well as cytokine related neurocircuitry alterations as defined by brain imaging. Treatment implications focus on the pathways by which inflammatory cytokines influence behavior, including therapeutic targets such as the inflammatory cytokines themselves, corticotropin-releasing hormone, and monoaminergic neurotransmitters and their precursors. Finally, recent data suggest that aggressive treatment strategies initiated before inflammation-inducing cancer treatments might prevent behavioral alterations, including depression, before they occur. PMID- 12893105 TI - HIV disease progression: depression, stress, and possible mechanisms. AB - There is much interest in whether depression and stress may explain the wide variability in the disease course of patients infected with human immunodeficiency virus (HIV). This article summarizes the large body of evidence examining whether depression and stress may have an impact on immune- and disease related parameters in HIV disease. Furthermore, we review what is known about the underlying biological mechanisms of HIV disease, such as alterations in glucocorticoids and catecholamines, which may help explain these psychoimmune relationships. Our review of the literature finds substantial evidence that chronic depression and stressful events may affect HIV disease progression. We know little, however, regarding the biological mechanisms that may account for these relationships. More research is warranted to investigate how depression and stress might impact HIV disease progression and what types of interventions might mitigate the negative impact of chronic depression and trauma. PMID- 12893106 TI - Prevalence, diagnosis, and pharmacological treatment of mood disorders in HIV disease. AB - Human immunodeficiency virus seropositive (HIV+) individuals are at a heightened risk of developing mood disorders and related syndromes. Over the past several decades, increased rates of mood disorders, including depression and mania, have been reported among HIV+ individuals. Because alterations in mood may impact on quality of life and perhaps reduce adherence to antiretroviral treatment regimens that are critical for preventing disease progression, recognition and effective treatment of mood disorders is essential. There are accumulating data showing that antidepressants and mood stabilizers, as well as other novel agents, might benefit HIV+ individuals suffering from a concomitant mood disturbance. This review highlights the relevant studies that have examined prevalence rates of mood disorders in HIV+ individuals, characteristics of HIV disease that influence the diagnosis and psychopharmacologic treatment of mood disorders, including complex interactions with antiretroviral medications, as well as the available evidence regarding the efficacy of agents used to treat depression and mania in the context of HIV disease. PMID- 12893107 TI - Relationship of depression to diabetes types 1 and 2: epidemiology, biology, and treatment. AB - This article reviews the rapidly accumulating literature on the relationship between mood disorders and diabetes mellitus. Recent studies have demonstrated that depression and its associated symptoms constitute a major risk factor in the development of type 2 diabetes and may accelerate the onset of diabetes complications. Since the mid-1980s, multiple longitudinal and cross-sectional studies have scrutinized the association of diabetes with depressive symptoms and major depression. Utilizing the search terms depressive disorders, psychiatry, diabetes, and pathophysiology in MEDLINE searches (1966-2003), this article reviews studies investigating pathophysiological alterations related to glucose intolerance and diabetes in depressed patients. The few randomized, controlled studies of treatment of depression in patients with diabetes are also described. Short-term treatment of depression in patients with diabetes improves their dysphoria and other signs and symptoms of depression. Future research will confirm whether response to psychotherapy and/or psychopharmacologic treatment improves glucose control, encourages compliance with diabetes treatment, and perhaps even increases longevity. PMID- 12893108 TI - Depression and obesity. AB - The prevalence of depression (10%) and overweight (65%) indicates that there is a probability that they will co-occur, but are they functionally related? This report used the moderator/mediator distinction to approach this question. Moderators, such as severity of depression, severity of obesity, gender, socioeconomic status (SES), gene-by-environment interactions and childhood experiences, specify for whom and under what conditions effects of agents occur. Mediators, such as eating and physical activity, teasing, disordered eating and stress, identify why and how they exert these effects. Major depression among adolescents predicted a greater body mass index (BMI = kg/m(2)) in adult life than for persons who had not been depressed. Among women, obesity is related to major depression, and this relationship increases among those of high SES, while among men, there is an inverse relationship between depression and obesity, and there is no relationship with SES. A genetic susceptibility to both depression and obesity may be expressed by environmental influences. Adverse childhood experiences promote the development of both depression and obesity, and, presumably, their co-occurrence. As most knowledge about the relationship between these two factors results from research devoted to other topics, a systematic exploration of this relationship would help to elucidate causal mechanisms and opportunities for prevention and treatment. PMID- 12893109 TI - Neuroimaging studies of mood disorder effects on the brain. AB - Studies of early-onset recurrent depression, late life depression associated with neurologic disorders, and bipolar illness have revealed structural brain changes within a neuroanatomical circuit. This circuit, originally described by, has been termed the limbic-cortical-striatal-pallidal-thalamic tract and is comprised of structures which are extensively interconnected. In three-dimensional magnetic resonance imaging studies of affective illness, many of the structures that comprise this tract have been found to have volume loss or structural abnormalities. Mechanisms proposed to explain volume loss in depression include glucocorticoid neurotoxicity, decreased brain-derived growth factor, decreased neurogenesis, and loss of plasticity. PMID- 12893110 TI - Depression in Alzheimer's disease: heterogeneity and related issues. AB - Several studies have demonstrated that the great majority of Alzheimer's disease (AD) patients suffer "noncognitive" neuropsychiatric symptoms. Depression is one of the most frequent neuropsychiatric comorbidities of AD. Affecting up to 50% of AD patients, depression in AD is associated with serious negative consequences for patients and their caregivers. Yet available studies on the natural course, etiology, and treatment of depression in AD have been few and equivocal. Heterogeneity in research methodology and etiology of depression in AD might have contributed to inconsistent findings across studies. Recently, the National Institute of Mental Health (NIMH) convened the Depression of Alzheimer's Disease Workgroup, which proposed provisional diagnostic criteria for depression of Alzheimer's Disease (NIMH-dAD). These criteria may provide a framework for future studies to clarify the unresolved issues in nosology, etiology, and treatment of depression in AD. A longitudinal cohort study of depression in incident AD cases may provide further syndrome refinement that would facilitate investigation of the etiology and treatment of depression in AD. PMID- 12893111 TI - Prevalence, etiology, and treatment of depression in Parkinson's disease. AB - Parkinson's disease (PD) is primarily a disease of elderly individuals with a peak age at onset of 55 to 66 years. It is characterized by bradykinesia, rigidity, tremor, and postural instability; and affects approximately 1 million individuals in the US and is the second most common neurodegenerative disease next to Alzheimer's disease. The motor symptoms of PD are the focus of pharmacotherapy, yet the nonmotor symptoms (e.g., dementia, psychosis, anxiety, insomnia, autonomic dysfunction, and mood disturbances) can be the most disturbing, disabling, and misunderstood aspects of the disease. Depressive symptoms occur in approximately half of PD patients and are a significant cause of functional impairment for PD patients. There is accumulating evidence suggesting that depression in PD is secondary to the underlying neuroanatomical degeneration, rather than simply a reaction to the psychosocial stress and disability. The incidence of depression is correlated with changes in central serotonergic function and neurodegeneration of specific cortical and subcortical pathways. Understanding comorbid depression in PD may therefore add to the understanding of the neuroanatomical basis of melancholia. PMID- 12893112 TI - Poststroke depression: prevalence, diagnosis, treatment, and disease progression. AB - In recent years, poststroke depression has attracted worldwide interest. This review focuses on the major research themes that have emerged. Pooled data from studies conducted throughout the world have found prevalence rates for major depression of 19.3% among hospitalized patients and 23.3% among outpatient samples. The diagnosis of poststroke depression is most appropriately based on a structured mental state exam and DSM-IV criteria for depression due to stroke with major depressive-like episode or depressive features. Rarely, poststroke patients may also develop bipolar mood disorder. The treatment of poststroke depression has been examined in several placebo-controlled randomized clinical trials with both nortriptyline and citalopram showing efficacy. The progression of recovery following stroke can be altered by treating depression, which has been shown to improve recovery in activities of daily living and cognitive impairment and to decrease mortality. In addition, two studies have demonstrated that poststroke depression can be prevented using antidepressant medication, which also decreases the frequency of associated physical illness. Furthermore, two studies have shown that premorbid depression can significantly increase the risk of stroke over the subsequent 10-15 years. The mechanisms underlying the association of cerebrovascular diseases and mood disorder are important areas for future investigation. PMID- 12893113 TI - Depression in epilepsy: prevalence, clinical semiology, pathogenic mechanisms, and treatment. AB - Depression is the most frequent comorbid psychiatric disorder in epilepsy. Its lifetime prevalence has been estimated at between 6% and 30% in population-based studies and up to 50% among patients followed in tertiary centers. The risk of suicide has been estimated to be 10 times higher than that in the general population. Although no one questions that epilepsy is a risk for depression, recent studies have also revealed that a history of depression is associated with a 4- to 6-fold greater risk of developing epilepsy. These data suggest either a possible "bi-directional" relationship between these two disorders or the presence of common pathogenic mechanisms that facilitate the occurrence of one in the presence of the other. The clinical presentation of depressive disorders in epilepsy can be identical to that of nonepileptic patients and can include major depression, bipolar and dysthymic disorders, and minor depression. In a significant percentage of cases, however, the clinical features of depression in epilepsy fail to meet any of the DSM-IV Axis I categories. Depression in epilepsy may be iatrogenically induced with various antiepileptic drugs used to treat the seizure disorder or after surgical treatment of intractable epilepsy. Despite its relatively high prevalence, depression remains unrecognized and untreated, and unfortunately its treatment is based on empirical and uncontrolled data. PMID- 12893114 TI - Persistent pain and depression: a biopsychosocial perspective. AB - This review highlights recent research findings on the relationship between persistent pain and depression and discusses the implications of these findings for future research in persons who suffer from both pain and depression. First, we briefly discuss advances in theories of pain that underscore the important role that depression can play in the chronic pain experience. Second, we discuss depression in persons suffering from chronic pain from a biopsychosocial perspective that takes into account both biological and psychosocial mechanisms linking pain and depression. Third, we address biomedical, psychosocial, and combined medical-psychosocial approaches to treatment in persons with persistent pain and depression. We conclude by highlighting future directions for research related to screening and diagnosis of depression in persons having persistent pain, treatment of comorbid pain and depression, and individual and subgroup differences in the experience of persistent pain and depression. PMID- 12893116 TI - Inductive bias strength in knowledge-based neural networks: application to magnetic resonance spectroscopy of breast tissues. AB - The integration of symbolic knowledge with artificial neural networks is becoming an increasingly popular paradigm for solving real-world applications. The paradigm provides means for using prior knowledge to determine the network architecture, to program a subset of weights to induce a learning bias which guide network training, and to extract knowledge from trained networks. The role of neural networks then becomes that of knowledge refinement. It thus provides a methodology for dealing with uncertainty in the prior knowledge. We address the open question of how to determine the strength of the inductive bias of programmed weights; we present a quantitative solution which takes the network architecture, the prior knowledge, and the training data into consideration. We apply our solution to the difficult problem of analyzing breast tissue from magnetic resonance spectroscopy (MRS); the available database is extremely limited and cannot be adequately explained by expert knowledge alone. PMID- 12893117 TI - A model for single and multiple knowledge based networks. AB - The inherent black-box nature of neural networks is an important drawback with respect to the problem of explanation of neural network responses. Although several articles have tackled the problem of rule extraction from a single neural network, just a few papers have investigated rule extraction from several combined neural networks. In this article we describe how to translate symbolic rules into the Discretized Interpretable Multi-Layer Perceptron (DIMLP) and how to extract rules from one or several combined neural networks. Our approach consists of characterizing discriminant hyperplane frontiers. Unordered rules are extracted in polynomial time with respect to the size of the problem and the size of the network. Moreover, the degree of matching between extracted rules and neural network responses is 100% on training examples. We applied single DIMLP networks to 17 data sets related to medical diagnosis and medical prognosis problems. Results based on 10-fold cross-validation showed that the DIMLP model was on average as accurate as standard multi-layer perceptrons (MLP). Furthermore, DIMLP networks were significantly more accurate than CN2 on eight problems, whereas only on one problem CN2 was better than DIMLP. Finally, a non Hodgkin lymphoma diagnosis problem based on classification of electrophoresis gels was defined. It turned out that ensembles of DIMLP networks were significantly more accurate than CN2 (96.1% +/- 1.4 versus 82.7% +/- 4.0). Finally, symbolic rules revealed the presence of five important spots for the discrimination of the class of Lymphocyte Leukemia/Chronic Lymphoid Leukemia (Lc/LLc), and the class of Centrocytic Lymphoma (Cc). PMID- 12893118 TI - Evolving connectionist systems for knowledge discovery from gene expression data of cancer tissue. AB - Microarray techniques have made it possible to observe the expression of thousands of genes simultaneously. They have recently been applied to study gene expression patterns in tissue samples. This may lead to highly desirable improvements in the diagnosis and treatment of human diseases. Statistical and machine learning methods have recently been used to classify cancer tissue based on gene expression data. Although some of these methods have achieved a high degree of accuracy, they generally lack transparency in their classification process. This, however, is crucial for the application in the medical field. In order to overcome this obstacle, we used knowledge-based neurocomputing (KBN), since KBN seeks to gain knowledge that is comprehensible to humans. In particular, we applied evolving fuzzy neural networks (EFuNNs) to classify cancer tissue, which is illustrated on the case studies of leukaemia and colon cancer. EFuNNs belong to the evolving connectionist system paradigm (ECOS) that has been recently introduced. They are well suited for adaptive learning and knowledge discovery. Fuzzy logic rules can be extracted from the trained networks and offer knowledge about the classification process in an easily accessible form. These rules point to genes that are strongly associated with specific types of cancer and may be used for the development of new tests and treatment discoveries. PMID- 12893119 TI - Explaining the output of ensembles in medical decision support on a case by case basis. AB - The use of ensembles in machine learning (ML) has had a considerable impact in increasing the accuracy and stability of predictors. This increase in accuracy has come at the cost of comprehensibility as, by definition, an ensemble model is considerably more complex than its component models. This is of significance for decision support systems in medicine because of the reluctance to use models that are essentially black boxes. Work on making ensembles comprehensible has so far focused on global models that mirror the behaviour of the ensemble as closely as possible. With such global models there is a clear tradeoff between comprehensibility and fidelity. In this paper, we pursue another tack, looking at local comprehensibility where the output of the ensemble is explained on a case by-case basis. We argue that this meets the requirements of medical decision support systems. The approach presented here identifies the ensemble members that best fit the case in question and presents the behaviour of these in explanation. PMID- 12893120 TI - Knowledge-based approach to septic shock patient data using a neural network with trapezoidal activation functions. AB - In this contribution we present an application of a knowledge-based neural network technique in the domain of medical research. We consider the crucial problem of intensive care patients developing a septic shock during their stay at the intensive care unit. Septic shock is of prime importance in intensive care medicine due to its high mortality rate. Our analysis of the patient data is embedded in a medical data analysis cycle, including preprocessing, classification, rule generation and interpretation. For classification and rule generation we chose an improved architecture based on a growing trapezoidal basis function network for our metric variables. Our results extend those of a black box classification and give a deeper insight in our patient data. We evaluate our results with classification and rule performance measures. For feature selection we introduce a new importance measure. PMID- 12893121 TI - Detection of change in shape: an advantage for concavities. AB - Shape representation was studied using a change detection task. Observers viewed two individual shapes in succession, either identical or one a slightly altered version of the other, and reported whether they detected a change. We found a dramatic advantage for concave compared to convex changes of equal magnitude. Observers were more accurate when a concavity along the contour was introduced, or removed, compared to a convexity. This result sheds light on the underlying representation of visual shape, and in particular the central role played by part boundaries. Moreover, this finding shows how change detection methodology can serve as a useful tool in studying the specific form of visual representations. PMID- 12893122 TI - People with Williams syndrome process faces holistically. AB - This study compared the performance of 47 adolescents and adults with Williams syndrome to 39 age-matched controls on a face recognition task. Using the whole part paradigm developed by Tanaka and his colleagues, we found that although performance overall was lower in the participants with Williams syndrome, both groups showed similar patterns of performance across the different conditions. Both groups performed significantly better in the whole-face than in the isolated part test condition for upright faces, but not for inverted faces. The whole-face advantage only in the upright condition provides strong evidence that people with Williams syndrome encode and recognize faces holistically in the same way as normal controls, suggesting the use of similar underlying neurocognitive mechanisms. These findings contradict earlier reports in the literature that people with Williams syndrome process faces abnormally. PMID- 12893123 TI - Limits on theory of mind use in adults. AB - By 6 years, children have a sophisticated adult-like theory of mind that enables them not only to understand the actions of social agents in terms of underlying mental states, but also to distinguish between their own mental states and those of others. Despite this, we argue that even adults do not reliably use this sophisticated ability for the very purpose for which it is designed, to interpret the actions of others. In Experiment 1, a person who played the role of "director" in a communication game instructed a participant to move certain objects around in a grid. Before receiving instructions, participants hid an object in a bag, such that they but not the director would know its identity. Occasionally, the descriptions that the director used to refer to a mutually visible object more closely matched the identity of the object hidden in the bag. Although they clearly knew that the director did not know the identity of the hidden object, they often took it as the referent of the director's description, sometimes even attempting to comply with the instruction by actually moving the bag itself. In Experiment 2 this occurred even when the participants believed that the director had a false belief about the identity of the hidden object, i.e. that she thought that a different object was in the bag. These results show a stark dissociation between an ability to reflectively distinguish one's own beliefs from others', and the routine deployment of this ability in interpreting the actions of others. We propose that this dissociation indicates that important elements of the adult's theory of mind are not fully incorporated into the human comprehension system. PMID- 12893124 TI - Children's reasoning about physics within and across ontological kinds. AB - Reasoning about seven physics principles within and across ontological kinds was examined among 188 5- and 7-year-olds and 59 adults. Individuals in all age groups tended to appropriately generalize what they learned across ontological kinds. However, children also showed sensitivity to ontological kind in their projections: when learning principles with reference to people they were more likely to assume that the principles apply to another person than to an inanimate object, and when learning with reference to an inanimate object they were more likely to assume that the principles apply to another inanimate object than to a person. Five-year-olds, but not 7-year-olds, projected concepts learned about people to a greater extent than principles learned about inanimate objects, closely paralleling the findings of Carey for the biological domain (Carey, S. (1985). Conceptual change in childhood. Cambridge, MA: MIT Press). Results from a separate sample of 22 5-year-olds suggest that the primary findings cannot be explained by response perseveration. The present findings indicate that children understand physics principles that apply to both animate and inanimate objects, but distinguish between these ontological kinds. PMID- 12893125 TI - Assignment of reference to reflexives and pronouns in picture noun phrases: evidence from eye movements. AB - Most structural Binding Theories predict a complementary distribution between reflexives and pronouns in picture noun phrases containing possessors (e.g. "Ken's picture of himself/him"). In two head-mounted eye-tracking experiments, listeners frequently violated Binding Theory predictions for reflexives, often interpreting the reflexives as taking an antecedent outside of the binding domain, and violating complementarity assumptions. Moreover, the pattern and timing of the eye movements showed Binding Theory violations for reflexives during the earliest moments of reference resolution. The results demonstrate that either binding constraints must be reformulated to decouple pronouns and reflexives or all reflexives in picture noun phrases must be treated as logophors, and thus exempt from structural Binding Theory. PMID- 12893126 TI - Numerosity discrimination in infants: evidence for two systems of representations. AB - Two experiments investigated numerosity discrimination in 6-month-old infants, comparing their performance on both large numbers (4 vs. 8 elements) and small numbers (2 vs. 4 elements) with both total filled area and total contour length controlled for. These studies provide the first direct comparison between discrimination of small and large numbers in infants with the same methodology, the same type of stimuli, and the same continuous variable controls. Results showed that infants succeeded in discriminating 4 from 8 elements but failed to discriminate 2 from 4 elements, providing evidence for the existence of two systems of number representations in infancy. PMID- 12893127 TI - Processing correlates of lexical semantic complexity. AB - This paper explores how verb meanings that differ in semantic complexity are processed and represented. In particular, we compare eventive verbs, which denote causally structured events, with stative verbs, which denote facts without causal structure. We predicted that the conceptually more complex eventive verbs should take longer to process than stative verbs. Two experiments, a lexical decision task and a self-paced reading study, confirmed this prediction. The findings suggest that (a) semantic complexity is reflected in processing time, (b) processing verb meanings involves activating properties of the event structure beyond participants' roles, and (c) more generally, lexical event structures, which subsume thematic roles, may mediate between syntactic knowledge and semantic interpretation. PMID- 12893128 TI - Eye contact does not facilitate detection in children with autism. AB - Eye contact is crucial in achieving social communication. Deviant patterns of eye contact behavior are found in individuals with autism, who suffer from severe social and communicative deficits. This study used a visual oddball paradigm to investigate whether children with high functioning autism have difficulty in detecting mutual gaze under experimental conditions. The results revealed that children with autism were no better at detecting direct gaze than at detecting averted gaze, which is unlike normal children. This suggests that whereas typically developing children have the ability to detect direct gaze, children with autism do not. This might result in altered eye-contact behavior, which hampers subsequent development of social and communicative skills. PMID- 12893129 TI - What is the magnitude of the subpopulation effect? AB - The effect of population subdivision on estimated match probabilities has been raised [Nature 339 (1989) 501; Am. J. Hum. Genet. 48 (1991) 819; Science 254 (1991) 1921]. Previous work [J. Forensic Sci. 39 (1994) 319; J. Forensic Sci. 39 (1994) 988; Am. J. Hum. Genet. 55 (1994) 533] has compared product rule estimates from differing databases and found that the "subpopulation" error may be of the order of a factor of 10. This approach compares an estimate with an estimate. This paper uses simulation to extend these studies by allowing a comparison to a 'true match probability' and supports the conclusion that subpopulation effects are mild. In addition the performance of recommendations 4.1 and 4.2 of NRC II [National Research Council and C.O.D.F. Science, The Evaluation of Forensic DNA Evidence, National Academy Press, Washington, DC, 1996]. PMID- 12893130 TI - Post-mortem SNP analysis of CYP2D6 gene reveals correlation between genotype and opioid drug (tramadol) metabolite ratios in blood. AB - Tramadol is an opioid drug metabolised in phase I by cytochrome P450 (CYP) enzymes, of which CYP2D6 is mainly responsible for the O-demethylation of tramadol, but is not involved in N-demethylation. Defects in the genes encoding drug metabolising enzymes (DMEs) may lead to adverse drug effects, even to death. To aid interpretation of the forensic toxicology results, we studied how the genetic variation of the CYP2D6 gene is reflected in tramadol metabolite ratios found in post-mortem samples. In 33 Finnish autopsy cases where tramadol was found, we analysed both the CYP2D6 genotype and the concentrations of tramadol and its metabolites O- and N-demethyltramadol. As expected, we found a correlation between the number of functional CYP2D6 alleles and the ratio of tramadol to O-demethyltramadol. We also found a correlation between the number of functional alleles and the ratio of tramadol to N-demethyltramadol. This can be explained by the complementary nature of the two main tramadol demethylation pathways. No known CYP2D6 inhibitors were associated with exceptional metabolic ratios. Furthermore, no accidental tramadol poisonings were associated with a defective CYP2D6 gene. Our results on the tramadol are among the first to demonstrate that genetic variation in drug metabolising enzymes can be analysed in post-mortem blood, and that it correlates well with the parent drug to metabolite ratios. The results also suggest that genetic factors play, in general, a dominant role over other factors in the metabolism of individual drugs. PMID- 12893131 TI - The use of cyclohexanone as a "derivatizing" reagent for the GC-MS detection of amphetamines and ephedrines in seizures and the urine. AB - A GC-MS method has been developed for the detection of amphetamine, methamphetamine, and the ephedrines, in seizures and the urine, based on on-GC condensation (derivatization) with cyclohexanone. The method is simple: the dried seizure material or the urine extract was mixed with cyclohexanone and injected into the GC-MS. The method was found to be superior to the methods based on acyl and trimethylsilyl (TMS) derivatization. Unlike for the acyl and TMS derivatives, the molecular and fragment ions of the cyclohexanone condensation products (cyclohexanone derivatives) were of substantial abundance, a useful property in unambiguous compound characterization. Furthermore, the high stability of the "derivatizing" reagent, cyclohexanone, compared with acyl and TMS derivatizing reagents, is a useful property in method development. The present method has proved selective and, tentatively, sensitive enough in the following areas (where methods based on acyl and TMS derivatization, as tested in this laboratory, have failed): (a) detection of amphetamine as a metabolite of methamphetamine; (b) detection of norpseudoephedrine as a metabolite of pseudoephedrine; (c) detection of amphetamine as an impurity of methamphetamine; (d) detection of cathine (norephedrine) as a constituent of Khat leaves; and (e) differentiation of Khat use from phenylpropanolamine use. PMID- 12893132 TI - Flies eggs: a new method for the estimation of short-term post-mortem interval? AB - Three modeling methods were used to estimate the time for laying of Lucilia sericata eggs (Diptera, Calliphoridae) after measurements of hatching times at several constant temperatures. These models were tested first under controlled conditions with 19 profiles of fluctuating temperatures, and on the other hand under field conditions on mice. All three models were allowed to determine the time of laying within a period of about 2h, and consequently this determines the time of death, as this species lays immediately after death under favorable conditions. Rearing of eggs sampled on a corpse may therefore contribute to the determination of a short post-mortem interval (PMI). PMID- 12893133 TI - Characterisation of variant alleles at the HumD21S11 locus implies unique Australasian genotypes and re-classification of nomenclature guidelines. AB - Several variant alleles of the HumD21S11 locus have only been reported in Australasian population samples. Fifteen such alleles were observed in Caucasian and Australian Aborigine sub-population databases compiled from residents of the state of Western Australia. Each variant was sequenced to authenticate the allelic designation and determine the structural conformation. Nine novel structural variants are described. The structure of the repeat region of these rare alleles combined with the STR designation brings aspects of the HumD21S11 nomenclature guidelines into question, in particular the designation of common incomplete repeats (or "0.2's"). The conformation of the sequences provides evidence in support of a genetic relationship between the Australian Aborigine and the Papuan people. PMID- 12893134 TI - Methamphetamine impurity profiling using a 0.32 mm i.d. nonpolar capillary column. AB - Classification of seized methamphetamine by impurity profiling can provide very useful information in criminal investigations of drug traffic routes, sources of supply and relationships between seizures. The aim of this study is to improve and develop an analytical method for detecting impurities such as starting materials and by-products in illegally prepared methamphetamine.HCl samples. A 50mg sample of methamphetamine.HCl was dissolved in 1 ml of buffer solution (four parts 0.1M phosphate buffer pH 7.0 and one part 10% Na2CO3). Impurities were extracted with 0.5 ml of ethyl acetate containing four internal standards (ISs) (n-decane, n-pentadecane, n-nonadecane and n-hexacosane) and analyzed by gas chromatography (GC) using a flame ionization detector (FID) on a DB-5 capillary column (0.32 mmi.d. x 30 m, film thickness 1.0 microm). The use of a middle-bore column offered better separation of the impurity peaks. The correction of the retention times of impurity peaks with four ISs made peak identification very accurate for subsequent data processing. Twenty-four characteristic peaks were selected for comparison and similarity and/or dissimilarity between samples, and the data were evaluated by the Euclidean distance of the relative peak areas after logarithmic transformation. The results indicate that the present method would be useful for methamphetamine impurity profiling. PMID- 12893135 TI - Different informativeness of the three hypervariable mitochondrial DNA regions in the population of Bologna (Italy). AB - Mitochondrial DNA (mtDNA) sequence variations at hypervariable regions HVI, HVII and HVIII were analysed in 100 unrelated Italians from Bologna. The values of the statistical parameters are in agreement with the range of European populations. We suggest that the less informative HVIII region may be useful to distinguish HVI-HVII identical sequences in forensic analysis especially when nuclear DNA cannot be investigated. PMID- 12893136 TI - Characterization of the explosive triacetone triperoxide and detection by ion mobility spectrometry. AB - The improvised explosive triacetone triperoxide (TATP) was synthesized and characterized by 1H-nuclear magnetic resonance (NMR), 13C-NMR, Raman and infrared (IR) spectroscopy. Triacetone triperoxide was subsequently analyzed by ion mobility spectrometry (IMS) in positive ion mode, and detected as a cluster of three peaks with a drift time of the most intense peak at 13.06 ms. Triacetone triperoxide was then analyzed after dissolution in toluene, where a dramatic increase in peak intensity was observed, at a flight time of 12.56 ms (K0=2.71 cm2V(-1)s(-1)). Triacetone triperoxide was subsequently analyzed by coupling the ion mobility spectrometer to a triple quadrupole mass spectrometer, where a single peak at m/z of 223 atomic mass units identified the species present in the ion mobility spectra as being triacetone triperoxide. PMID- 12893137 TI - Allele frequencies for 15 STR loci in Van-Agri districts of the Eastern Anatolia region of Turkey. AB - Allele frequencies of 13 tetrameric short tandem repeat (STR) loci (D3S1358, TH01, D21S11, D18S51, D5S818, D13S317, D7S820, D16S539, CSF1PO, vWA, D8S1179, TPOX, FGA) and 2 pentameric short tandem repeat loci (Penta E and Penta D) included in the PowerPlex 16 kit were obtained from a sample of 116 unrelated individuals in Van-Agri districts of the Eastern Anatolia region of Turkey. The expected performance of these loci for personal identification and paternity testing in this population was estimated. PMID- 12893138 TI - Genetic profile of the Ecuadorian Mestizo population (Ecuador-South America) by using the Power Plex 16 System Kit. AB - Allele frequency data for the 15 STR systems and Amelogenine were determined in a population sample of healthy Mestizo unrelated individuals. All loci met Hardy Weinberg expectations and the high discrimination power of combined system showed the forensic efficiency of these genetic markers. There is a lack of information on Ecuadorian population from a genetic point of view and therefore no previous publications on the distribution of STRs on Mestizos is available. Mestizos are descendents of Spanish and Amerindian people, however, significant differences were found between the former and the populations from Spanish peninsula, that have been analyzed for these genetic markers. PMID- 12893139 TI - Genetic data on 12 STRs (F13A01, F13B, FESFPS, LPL, CSF1PO, TPOX, TH01, vWA, D16S539, D7S820, D13S317, D5S818) from four ethnic groups of Sao Paulo, Brazil. AB - Allelic frequencies for 12 short tandem repeats (STRs) (F13A01, F13B, FESFPS, LPL, CSF1PO, TPOX, TH01, vWA, D16S539, D7S820, D13S317 and D5S818) were estimated, also as forensic parameters, from a sample of 916 unrelated Brazilian subjects classified into four ethnic groups: European-derived, African-derived, Brazilian Mulattos and Asian-derived. PMID- 12893140 TI - Population data of nine STR loci, D3S1358, vWA, FGA, TH01, TPOX, CSF1PO, D5S818, D13S317 and D7S820, in Bangladeshis and Indonesians. AB - Allele frequencies of nine short tandem repeat (STR) loci, D3S1358, vWA, FGA, TH01, TPOX, CSF1PO, D5S818, D13S317 and D7S820, were determined for 127 unrelated Bangladeshi individuals and 105 unrelated Indonesian individuals using the AmpFLSTR Profiler Kit. The genotype frequency distributions of the nine STR loci were in the Hardy-Weinberg equilibrium for both populations. PMID- 12893141 TI - STR frequencies for the PowerPlex 16 System Kit in a population from Northeast Spain. AB - The aim of this paper, was to obtain the frequencies for the 15 STR loci included in the PowerPlex 16 System Kit on a population of 204 unrelated Caucasian individuals living in Northeast of Spain in order to use for forensic purposes. Statistical analyses were performed using the programs GENEPOP version 3.3 and Powerstats version 1.2. The results showed that all the loci met Hardy-Weinberg expectations. PMID- 12893142 TI - Anatomy and biomechanics of the medial patellofemoral ligament. AB - The medial patellofemoral ligament (MPFL) is a band of retinacular tissue connecting the femoral medial epicondyle to the medial edge of the patella. The MPFL is approximately 55 mm long, and its width has been reported to range from 3 to 30 mm. The MPFL is overlaid by the distal part of vastus medialis obliquus to a variable extent, and fibres of MPFL merge into the deep aspect of the muscle. Despite the MPFL being very thin, it had a mean tensile strength of 208 N, and has been reported to be the primary passive restraint to patellar lateral displacement. Lateral patellar displacement tests in vitro showed that the patella subluxed most easily at 20 degrees knee flexion. The contribution of the MPFL to resisting patellar lateral subluxation was greatest in the extended knee. This finding was linked to the retinaculae being tightest in full knee extension, and slackening with flexion. PMID- 12893143 TI - The anatomy and reconstruction of the medial patellofemoral ligament. AB - INTRODUCTION: Reconstruction of the MPFL for recurrent and acute lateral dislocation of the patella is becoming more accepted. This study aimed to further define the medial patellofemoral ligament (MPFL) anatomy and to describe the most suitable sites for graft attachment when reconstructing the MPFL. MATERIALS AND METHODS: Anatomical dissection was performed on 25 embalmed knee specimens. Four separate specimens were used to assess graft fixation points. Sites were tested for isometricity from 0 to 120 degrees of knee flexion. Length patterns were considered isometric if there was less than 5 mm of length change throughout range of motion. RESULTS: The MPFL attaches from the posterior part of the medial epicondyle, to the superomedial patella, the under-surface of vastus medialis, and the quadriceps tendon. Optimal attachment points for an MPFL graft were the superior patella and the femoral attachment points of the MPFL (just distal to the adductor tubercle) and points posterior and inferior to this. CONCLUSION: We have defined the anatomy and kinematics of the MPFL with reconstruction of this ligament in mind. PMID- 12893144 TI - Regeneration of degenerated articular cartilage after high tibial valgus osteotomy for medial compartmental osteoarthritis of the knee. AB - The aim of the current study was to document regeneration of the articular cartilage after correction of varus deformity for osteoarthritis of the knee. The repair of articular cartilage after high tibial valgus osteotomy for medial compartmental osteoarthritis was observed in 146 knees of 115 patients. The mean age of the patients at osteotomy was 65+/-7 years (range 47-80). Observations were made through an arthrotomy at the time of removal of the blade plate, an average of 2 years after the initial osteotomy. Regeneration of the degenerated cartilage was classified into three stages. There were no regenerative findings (Stage A) in 13 knees; partial regeneration with fibrocartilaginous tissue (Stage B) in the previous degenerated area (Stage B) in 86 knees; and total coverage by new regenerated fibrocartilage or hyaline-like cartilage (Stage C) in 47 knees. Mature regeneration (Stage C) was observed more frequently in the knees with increased width of the medial joint space after high tibial osteotomy than in those with unchanged width (P<0.01), and in the knees with more than 5 degrees of anatomical valgus angulation after osteotomy than in those with less than 5 degrees (P=0.05). Even the knees with eburnation of subchondral bone in the weight-bearing portion showed Stage C regeneration in 26 out of 80, in contrast with no regeneration in two knees with degenerated cartilage remaining. O'Driscoll's histological and histochemical grading score was higher in the Stage C group (P<0.02). The weight-bearing portion of the medial femoral condyle with exposed bone was concluded to be partially or entirely covered by newly regenerated cartilage after 1 year after adequate correction of varus deformity by high tibial osteotomy without antologous chondrocyte implantation. PMID- 12893145 TI - Varus/valgus alignment of the femoral component in total knee arthroplasty. AB - The position of the femoral component in 362 total knee replacements was assessed radiologically. A subgroup of 32 knees, 18 of which had perfect alignment and 14 with imperfect alignment underwent CT scout scans of the femur from which the mechanical axis of the femur could be measured. Radiologically 92% of all components were implanted within 3 degrees of the target value. There was close correlation between the CT and X-ray measurements in the subgroup. Deviation from the mechanical axis was 1.16 degrees (range -2.5 to +2) in the perfectly aligned knees, validating both surgical technique and radiological assessment. Causes for inaccuracy in femoral placement and future likely developments are discussed. PMID- 12893146 TI - Varus/valgus alignment of the tibial component in total knee arthroplasty. AB - The varus/valgus alignment of the tibial components of 350 total knee arthroplasties was assessed radiologically. All the tibial components were implanted using an extramedullary guide under the supervision of the senior author. Of components, 96.3% were implanted within 2 degrees of the perpendicular to the longitudinal axis of the tibia. In order to validate our X-ray assessment, a subgroup of 40 knees was re-assessed using a CT scanogram. Analysis of this subgroup showed a close correlation between the results using the two different methods (mean difference 0.88 degrees, S.D. 0.75). We believe that with anticipated future advances in robotic and computer assisted surgery it is important to set the benchmark for what can be achieved with current technology. PMID- 12893147 TI - Computer based method for the three-dimensional kinematic analysis of combined posterior cruciate ligament and postero-lateral complex reconstructions on cadaver knees. AB - The aim of this study is to evaluate the effect of combined posterior cruciate ligament (PCL) and postero-lateral corner (PLC) reconstruction on laxity and three-dimensional kinematics of cadaver knees. We performed anatomical double bundle PCL reconstruction, and functional one bundle 'over-the-bottom' PCL reconstruction combined with one type of PLC reconstruction, running from the postero-lateral tibia to an isometric point near the lateral epicondyle of the femur. Our results showed that combined reconstruction was necessary to restore rotatory laxity. PLC reconstruction, according to the technique described, invariably created a shift towards internal rotation of the kinematic curves, compared to the intact knee. PMID- 12893148 TI - Drilling from the intercondylar area for treatment of osteochondritis dissecans of the knee joint. AB - We demonstrate a new method in which a drilling is made from the intercondylar space, and its efficacy in treating osteochondritis dissecans (OCD) of the knee in skeletally immature patients with relatively stable lesions with an intact articular surface, in cases where there was failure of initial non-operative management. The lesions of 16 knees of 12 patients with OCD of the femoral condyles failed to heal by conservative treatment for more than 3 months (average 5-6 months) and thereafter were arthroscopically treated with drilling from not the transarticular but the intercondylar bare area without damaging the articular surface. Eight lesions involved the medial femoral condyle, and eight involved the lateral femoral condyle. The average follow-up was 16 months. All lesions healed after drilling, and the average time of healing was 4 months by X-ray and 7 months by MRI. The average Lysholm score improved from an average of 70.4 points at preoperation to an average of 97.8 points after operation. The results of the Hughston Rating Scale were similar: 10 of the 12 patients showed excellent results and the remaining two patients good results. We advocate our new and less invasive procedure of drilling from the bare area of the intercondylar space for OCD in the knee joint of skeletally immature patients who have had failure of initial non-operative management. PMID- 12893149 TI - Biomechanics of passive knee joint in drawer: load transmission in intact and ACL deficient joints. AB - A non-linear 3D finite element model of the passive human tibiofemoral knee joint consisting of two bony structures and their articular cartilage layers, menisci, and four principal ligaments was used to investigate the detailed response of the unconstrained joint under up to 100 N posterior femoral force at different flexion angles from 0 to 90 degrees. The analysis was repeated after the transection of the anterior cruciate ligament (ACL). The boundary conditions were selected to assure a stable and unconstrained response of the joint throughout the range of motion. The results indicated the ACL as the primary structure to resist the drawer load throughout the range of flexion considered and that the joint primary and coupled laxities substantially increased in its absence. At full extension under drawer, forces in collateral ligaments increased significantly resulting in larger overall contact forces as the ACL was transected. In the ACL-deficient joint, such large forces in collateral ligaments, however, diminished as flexion angle varied from 0 to 90 degrees. At full extension or flexion angles up to approximately 30 degrees, the medial meniscus and adjacent medial tibial and femoral cartilage layers were subjected to substantially larger loads and stresses following the transection of the ACL. Adequate consideration of such couplings is important in avoiding further damage to joint structures subsequent to an injury and restoring adequate function following injuries to primary components. PMID- 12893150 TI - Synergy of medial and lateral hamstrings at three positions of tibial rotation during maximum isometric knee flexion. AB - Rotation of the knee has been used to isolate the strength of the medial and lateral hamstrings during manual testing of the knee flexors. The purpose of this study was to determine if medial and lateral rotation of the knee during manual knee flexor strength testing increased the electromyographic activity of the respective hamstrings. Twenty-three women between 22 and 36 years old with no history of lower extremity injury or disease participated in the study. Indwelling fine wire electrodes were used to record EMG activity of the medial (semitendinosus and semimembranosus) and lateral (long and short heads of the biceps femoris) hamstring muscles during maximally resisted knee flexion with neutral, medial, and lateral rotation of the knee. Repeated measures analysis of variance with post hoc Bonferroni adjustments were used to compare EMG activity across the three tests. EMG activity increased significantly for the target hamstrings during ipsilateral rotation (P<0.05). The semitendinosus had a mean activity of 109% Max. during medial rotation as opposed to 95% Max. during lateral rotation. The semimembranosus averaged 107 and 89% Max. in medial and lateral rotation respectively. Conversely, both the long and short head of the biceps muscle showed significantly higher activity (P<0.05) during lateral compared to medial rotation (110 and 108% compared to 93 and 97%, respectively). Even though the differences are statistically significant they ranged from 2 to 13% only of maximum activity, the clinical importance of this small change in EMG activity is questionable. PMID- 12893151 TI - Arthroscopically assisted ACL reconstruction. Is a drain necessary? AB - We report on the results of a prospective randomised controlled trial to evaluate the use of an intra-articular drain following arthroscopically assisted ACL reconstruction using patellar tendon autograft. Forty-nine patients were recruited for the trial and randomised into receiving or not receiving a drain following surgery. An independent observer who was blinded for the use of the drain assessed the knees for swelling, range of movement (ROM) and pain at 2, 4 and 6 weeks postoperatively. Muscle strength was assessed at 12 weeks following surgery using the KIN COM III isometric dynamometer. We found that the knees that were drained following surgery initially had less swelling and a better ROM. However, at 4 weeks this difference had disappeared. At 3 and 6 months, there was no functional difference between the two groups. We recommend that no drain be used following ACL reconstruction as removal of the drain is uncomfortable and carries theoretical and avoidable risks. PMID- 12893152 TI - Is bilateral knee arthroscopy suitable as a day-case procedure? AB - A retrospective analysis of 100 out of 108 cases has been audited for suitability of bilateral knee arthroscopy as a day-case procedure. Fifty-three female and 47 male patients with age range of 15-78 years underwent bilateral knee arthroscopy. All patients had general anaesthetic and were ASA grade 1 or 2. Eight patients had more than one diagnosis and procedures. Post-operatively 68% could walk without walking aids, 1% had 1 stick and 31% needed 2 sticks and were given elbow crutches. One patient stayed overnight due to poor pain control and 1 attended A&E due to effusion. There were 5 (2.5%) major and 62 (31%) minor complications in 200 knees. Mild post-operative pain was the commonest minor complication in 52 (26%) out of 62 knees and was attributed to high number of bilateral arthritis cases in this study. Complication rate of bilateral arthroscopy is similar to unilateral arthroscopy. It is concluded that bilateral arthroscopy is safe as a day-case procedure. PMID- 12893153 TI - A prospective randomised trial comparing intra-articular Hyalgan injection and arthroscopic washout for knee osteoarthritis. AB - Thirty-eight patients with symptomatic knee osteoarthritis without mechanical symptoms were randomised after informed consent to receive either a course of intra-articular Hyalgan injections or an arthroscopic washout. The patients were prospectively assessed pre-intervention, 6 weeks, 3 months, 6 months and 1 year using a 10 cm visual analogue pain score, the Knee Society function score and the Lequesne index. There was no significant difference between the two groups at 6 weeks, 3 months, 6 months or 1 year. The use of intra-articular Hyalgan injections in patients with knee osteoarthritis without mechanical symptoms gives results comparable with arthroscopic washout. Hyalgan is an alternative to arthroscopy in this patient group. Further study is needed to confirm these findings and improve patient selection. PMID- 12893154 TI - Bone marrow edema syndrome of the knee: a case report and review of the literature. AB - The authors present a 45-year-old man with bone marrow edema syndrome (BMES) of the medial condyle of the knee. Early diagnosis of BMES was based upon clinical, bone scintigraphy and magnetic resonance (MRI) findings. After failure of the conservative treatment, core decompression was performed. Histological examination showed no evidence of osteoporosis or osteonecrosis. Within 2 months after the operation, the patient was pain-free and he resumed his previous activities. At 1-year follow-up examination, MRI showed complete resolution of all signal abnormalities; at 3-year follow-up the patient remained symptom-free and with no functional limitations. PMID- 12893157 TI - Trace element, toxin and drug elimination in hair with particular reference to the horse. PMID- 12893158 TI - Noxious stimulation during orthopaedic surgery results in EEG 'arousal' or 'paradoxical arousal' reaction in isoflurane-anaesthetised sheep. AB - The effects of surgical stimuli on haemodynamic and electroencephalographic (EEG) variables were determined in 25 adult ewes undergoing an experimental orthopaedic procedure in isoflurane anaesthesia. Data were recorded after 15 min of constant end-tidal concentration of approximately 2.2% isoflurane (SS: steady state=baseline), during skin disinfection (DIS), incision (INC), drilling of the first hole through the tibia (DRI) and insertion of a threaded pin (PIN) for external fixation. Stimulation resulted in a significant increase in systolic and mean arterial pressure above SS at INC, DRI and PIN. Haemodynamic changes were accompanied by either significant increases or decreases in EEG median frequency (MF) and 80% spectral edge frequency (SEF80) above or below SS at all four stimulation time points suggesting 'arousal' or 'paradoxical arousal' reaction, respectively. We conclude, that either type of EEG activation pattern could be elicited dependent on stimulation intensity and level of anaesthetic depth. PMID- 12893159 TI - Biochemical characterisation of navicular hyaline cartilage, navicular fibrocartilage and the deep digital flexor tendon in horses with navicular disease. AB - The study hypothesis was that navicular disease is a process analogous to degenerative joint disease, which leads to changes in navicular fibrocartilage and in deep digital flexor tendon (DDFT) matrix composition and that the process extends to the adjacent distal interphalangeal joint. The objectives were to compare the biochemical composition of the navicular articular and palmar cartilages from 18 horses with navicular disease with 49 horses with no history of front limb lameness, and to compare navicular fibrocartilage with medial meniscus of the stifle and collateral cartilage of the hoof. Cartilage oligomeric matrix protein (COMP), deoxyribonucleic acid (DNA), total glycosaminoglycan (GAG), metalloproteinases MMP-2 and MMP-9 and water content in tissues were measured. Hyaline cartilage had the highest content of COMP and COMP content in hyaline cartilage and tendon was higher in lame horses than in sound horses (p<0.05). The concentration of MMP-2 amount in hyaline cartilage was higher in lame horses than in sound horses. The MMP-2 amounts were significantly higher in tendons compared to other tissue types. Overall, 79% of the lame horses with lesions had MMP-9 in their tendons and the amount was higher than in sound horses (p<0.05). In horses with navicular disease there were matrix changes in navicular hyaline and fibrocartilage as well as the DDFT with potential implications for the pathogenesis and management of the condition. PMID- 12893160 TI - Ultrasound imaging of the canine tympanic bulla. AB - A procedure for imaging the canine tympanic bulla, external ear canal and adjacent structures using currently available ultrasound equipment was established. Lateral and ventral transducer positions were identified for this purpose and a 6.5 MHz curvilinear transducer was considered to be optimal. The sonographic appearance of these structures in cadavers and live dogs unaffected by ear disease is documented. Fluid was introduced into the tympanic bullae of the cadavers and its presence could be identified through the bony wall of the bulla. The ability of ultrasound to differentiate between gas and fluid within the bulla has important clinical implications as this is a common occurrence in dogs with middle ear disease. Ultrasound has several advantages over other imaging modalities and the examination procedure was well tolerated by unsedated dogs. PMID- 12893161 TI - Piecemeal degranulation (PMD) morphology in feline circulating eosinophils. AB - Buffy coat preparation from six cats with 600-4560 circulating eosinophils/microL was collected by either blood centrifugation or sedimentation, fixed in 2.5% glutaraldehyde, post-fixed in either 1% osmium or in 1.5% potassium ferrocyanide reduced osmium, ultra-sectioned and examined by transmission electron microscopy. Ultrastructural changes of piecemeal degranulation (PMD), which is a mechanism of eosinophil granule contents release indicative of eosinophil activation, were observed in specific granules from all the samples examined. The spectrum of PMD included coarsening of the granule matrix, budding vesicles, fragmented granule cores and lucent granules. The number of presumably activated eosinophils with ultrastructural evidence of PMD did not correlate with the level of eosinophilia. The lack of correlation suggested that, analogously with humans, blood eosinophil count might not represent the best criterion to evaluate the contribution of eosinophils to tissue damage in certain feline eosinophil-associated diseases. PMID- 12893162 TI - Lipopolysaccharide and interferon gamma activate nuclear factor kappa B and induce cyclo-oxygenase-2 in equine vascular smooth muscle cells. AB - Equine endotoxaemia is an important cause of morbidity and mortality in horses caused by the interaction of bacterial lipopolysaccharide (LPS) with cells such as macrophages and vascular smooth muscle. In this study we isolated equine vascular smooth muscle from a variety of vessels and stimulated it with LPS and human interferon (hIFN)-gamma. Using reverse transcriptase polymerase chain reaction (rt-PCR) and Western blot analysis we show that cyclooxygenase-2 (COX-2) is readily expressed in equine vascular smooth muscle. Vascular smooth muscle cells produced prostaglandin E2 in response to LPS and hIFNgamma. Using similar approaches we saw very limited expression of inducible nitric oxide synthase (iNOS) in only one vascular smooth muscle preparation. LPS and IFNgamma caused translocation of the transcription factor nuclear factor kappa B (NfkappaB) to the nucleus in equine cells suggesting the limited iNOS production seen in our cells is not due to deficits in this signal transduction pathway. These data suggest that in equine vascular smooth muscle COX-2 and NfkappaB are likely to play important roles in the pathogenesis of equine endotoxaemia. PMID- 12893163 TI - Effects of poppy seed meal on egg production and hatching results in quail (Coturnix coturnix japonica). AB - This study was carried out to determine the effects of poppy seed meal (PSM) on the egg production and hatching of quail. Two experiments were undertaken. In the first experiment, 240 quail, reared under the same conditions, were divided into four groups of 60 birds each (which were, in turn, subdivided into four subgroups of 15 each for experimental replicates). The first, second, and third experimental groups were fed ratios containing 5%, 15%, and 25% PSM, respectively, over 12 weeks. The control group received no PSM. Feed and water were supplied ad libitum and artificial light was provided for 16 h a day. During the first experiment, egg production, egg weight, and feed conversion ratio (FCR) were measured. In the second experiment, 240 female and 48 male quail were divided into four groups, as in the preceding experiment, and the birds were housed in cages at a ratio of one male to five females. The diets and their presentation to the quails were the same as in the preceding experiment. The results of the two experiments showed that the addition of PSM to the feed ratios increased egg production, feed consumption, and feed conversion per kg of eggs and per dozen eggs. However, PSM addition at all percentages significantly reduced (P<0.001) hatchability, mainly because of increased infertility and embryonic deaths. PMID- 12893164 TI - Effect of dietary fat source and exercise on odorant-detecting ability of canine athletes. AB - Eighteen male English Pointers (2-4 years of age, 23.94+/-0.54 kg body weight) were allotted to three diet and two physical conditioning groups to evaluate the effect of level and source of dietary fat on the olfactory acuity of canine athletes subjected to treadmill exercise. Diet groups (6 dogs/diet) consisted of commercially prepared diets (minimum of 26% crude protein) containing 12% fat as beef tallow (A), 16% fat provided by equivalent amounts of beef tallow and corn oil (B), or 16% fat provided by equivalent amounts of beef tallow and coconut oil (C). This dietary formulation resulted in approximately 60% of the total fatty acid being saturated for diets A and C, while approximately 72% of the total fatty acids were unsaturated in diet B. One-half of the dogs within each dietary group were subjected to treadmill exercise 3 times per week for 30 min (8.05 km/h, 0% grade) for 12 weeks. All dogs were subjected to a submaximal exercise stress test (8.05 km/h, 10% slope for 60 min) every four weeks beginning at week 0. Olfactory acuity was measured utilizing behavioral olfactometry before and after each physical stress test. Non-conditioned (NON) dogs displayed a greater decrease (P<0.05) in olfactory acuity following exercise, while physically conditioned (EXE) dogs did not show a change from pre-test values. A diet by treatment interaction (P<0.10) was detected over the course of the study. NON dogs fed coconut oil had decreased odorant-detecting capabilities when week 4 values were compared with week 12 values. Feeding a diet that is predominately high in saturated fat may affect the odorant-detecting capabilities of working dogs. Additionally, these data indicate that utilization of a moderate physical conditioning program can assist canine athletes in maintaining olfactory acuity during periods of intense exercise. PMID- 12893165 TI - Sources of sampling variation in saliva cortisol in dogs. AB - The main advantage of collecting saliva cortisol as opposed to plasma cortisol is that it is non-invasive and therefore it is now widely used in stress measurement studies on farm animals and dogs. Although a plasma cortisol response to handling associated with blood collection generally occurs at 3 min from the commencement of handling, there is no information in the literature on the time course of the response of salivary cortisol concentration to handling. The aims of these experiments were to (1). determine if there is a response to up to 4 min handling that affects cortisol concentration in saliva and (2). determine the main causes of variation in saliva cortisol in dogs over time. In experiment 1, saliva was collected from six Kelpies at 0 min then 2, 3 or 4 min after the commencement of restraint. There was no handling effect found in up to 4 min sampling time. In experiment 2, saliva was collected from six Labrador Retrievers five times in 2 h (14:00-16:00), three days a week for four weeks. Some of the sources of variation in saliva cortisol over time included between dog variation that varied over a period of days and variation between occasions that affected the group of dogs as a whole. PMID- 12893166 TI - Genome re-arrangements associated with loss of pathogenicity of the gamma herpesvirus alcelaphine herpesvirus-1. AB - The alcelaphine herpesvirus 1 (AlHV-1) causes malignant catarrhal fever in ruminants. Previous work had shown that serial passage of AlHV-1 in culture resulted in genome alterations that are associated with a loss in pathogenicity. Here we have analysed the re-arrangements that occur in more detail. None of the observed re-arrangements was entirely consistent. However, they did all involve translocation of a similar region of DNA from around the centre of the genome to areas either next to or in between terminal repeat elements at either end of the genome. There was also a concomitant loss of the wild-type locus. These re arrangements appeared to be associated with the loss of virulence and the appearance of cell-free virus. PMID- 12893167 TI - Histone modifications. PMID- 12893168 TI - Protein mass analysis of histones. AB - Posttranslational modification of chromatin-associated proteins, including histones and high-mobility-group (HMG) proteins, provides an important mechanism to control gene expression, genome integrity, and epigenetic inheritance. Protein mass analysis provides a rapid and unbiased approach to monitor multiple chemical modifications on individual molecules. This review describes methods for acid extraction of histones and HMG proteins, followed by separation by reverse-phase chromatography coupled to electrospray ionization mass spectrometry (LC/ESI-MS). Posttranslational modifications are detected by analysis of full-length protein masses. Confirmation of protein identity and modification state is obtained through enzymatic digestion and peptide sequencing by MS/MS. For differentially modified forms of each protein, the measured intensities are semiquantitative and allow determination of relative abundance and stoichiometry. The method simultaneously detects covalent modifications on multiple proteins and provides a facile assay for comparing chromatin modification states between different cell types and/or cellular responses. PMID- 12893169 TI - Measurement of histone acetyltransferase and histone deacetylase activities and kinetics of histone acetylation. AB - Dynamic histone acetylation has a role in chromatin remodeling and in the regulation of transcription. Histone deacetylases (HDACs) and histone acetyltransferases (HATs) catalyze reversible histone acetylation. HATs and HDACs exist as multiprotein complexes that have coactivator and corepressor activities, respectively. The steady-state level of acetylation at a chromatin site is determined by the local net activities of these enzymes. Here we describe methods to isolate different subcellular fractions (cytosol, nuclei, tightly bound nuclear, loosely bound nuclear, immunoprecipitated multiprotein complexes, and nuclear matrix) to determine the subcellular distribution of HAT and HDAC activities. Procedures to assay the activities of these enzymes and to measure the kinetics of histone acetylation and deacetylation are presented. PMID- 12893170 TI - Expression, purification, and analysis of MOZ and MORF histone acetyltransferases. AB - Histone acetylation is one major mechanism by which chromatin structure and function are regulated. Besides histones, many nonhistone proteins are also acetylated in vivo. Aberrant acetylation has been linked to the development of various human diseases. Through acetylating histone and nonhistone proteins, histone acetyltransferases (HATs) play fundamental roles in regulating chromatin remodeling, transcription, and other nuclear processes. Known HATs belong to several groups, including the GCN5/PCAF, p300/CBP, and MYST families. ESA1, SAS3, MOF, TIP60, HBO1, MOZ, and MORF are the MYST family members with demonstrated HAT activity. The MOZ and MORF genes are rearranged by chromosome abnormalities associated with several types of leukemia, so these two HATs have been implicated in leukemogenesis. Compared with p300, CBP, and PCAF, much less is known about MOZ and MORF. To elucidate the function and regulation of these two interesting HATs, we have conducted their initial characterization. Here we describe the expression, purification, and activity analysis of MOZ and MORF. For comparison, we also include the procedure for expression and purification of PCAF. These methods are useful not only for functional characterization of MOZ, MORF, PCAF, and other HATs, but also for preparation of HAT proteins to screen compound libraries and obtain inhibitors with potential therapeutic value. PMID- 12893171 TI - Enzymatic assays for NAD-dependent deacetylase activities. AB - The NAD-dependent deacetylases are a new class of enzymes responsible for the removal of acetyl groups from lysines on proteins. Instead of water, the NAD dependent deacetylases use a highly reactive ADP-ribose intermediate as a recipient for the acetyl group. The products of the reaction are nicotinamide, acetyl-ADP-ribose, and a deacetylated substrate. Many assays have been developed for the measurement of NAD-dependent deacetylase activity. In this review we present assays based on each of the two reactions catalyzed by these enzymes, deacetylation and NAD hydrolysis. First we describe methods for the production of acetylated protein and peptide substrates for use in deacetylation reactions. Then we describe four methods for assaying deacetylation, three of which directly measure the loss of acetyl groups from a protein or peptide substrate, and one that measures acetate production. We also describe two indirect methods for following enzyme activity, NAD hydrolysis and a novel NAD-nicotinamide exchange reaction. Finally, a quantitative method using a monoacetylated peptide as a substrate and HPLC to measure products is described. PMID- 12893172 TI - Histone phosphorylation: how to proceed. AB - Among all posttranslational modifications that occur on histone tails, phosphorylation is the one that establishes a direct link between chromatin remodeling and intracellular signaling pathways. Specific, conserved serine residues are present on the N-terminal tails of each histone. These are phosphoacceptor sites for a number of kinases, whose identification is essential to decipher the transduction routes leading to various physiological responses. In the case of histone H3, phosphorylation at the Ser10 residue may lead to either activated gene expression or chromatin condensation during mitosis. In addition, phosphorylation at specific sites may be coupled to other distinct modifications, such as acetylation and methylation, generating the so-called "histone code" which postulates that well defined combinatorial modifications at histone tails correspond to specific physiological responses. Here we describe a number of experimental methodologies that are essential for the study of histone phosphorylation. While chromatin immunoprecipitation is useful in recognizing gene targets, the in-gel kinase assay is a first, essential step in establishing the identity of the kinase(s) that operates in response to a specific signaling pathway. The subsequent use of in vitro kinase assays is helpful in validating the implication of a candidate kinase. These powerful approaches are important as identification of the signaling transduction routes leading to chromatin remodeling is critical to an understanding of all cellular processes. PMID- 12893173 TI - Methods and tips for the purification of human histone methyltransferases. AB - Recently developed biochemical techniques have enabled researchers to study histone modifications more easily and accurately. One of these modifications, histone lysine methylation, has been shown to be highly stable and to represent an epigenetic alteration. Extensive biochemical analyses have led to discoveries about the nature and functions of this modification, thus accelerating our understanding of this crucial epigenetic event. Here we describe basic methods for purification and biochemical analysis of lysine-directed, histone methyltransferases from HeLa cell-derived extracts. In the section on substrate preparation, we describe a simple method for the preparation of recombinant substrates, although we recommend using native substrates for initial detection of the activities. The purification protocols for several histone methyltransferases have been streamlined so that those researchers with a basic understanding of biochemistry can perform them. We also describe many tips and provide suggestions to avoid common pitfalls in the biochemical analysis of histone methyltransferases. PMID- 12893174 TI - In vivo assays to study histone ubiquitylation. AB - The importance of histone acetylation, phosphorylation, and methylation in transcription and other DNA-mediated processes is now well established. Histones are also ubiquitylated, but in contrast to the majority of ubiquitylated proteins, ubiquitylated histones are not generally targeted for degradation and may play roles similar to those of other histone modifications. Antibodies against acetylated histones have provided unique insights into the regulation, distribution, and cellular roles of these modified histones. In this report, we describe methods to identify ubiquitylated histones in budding yeast and HeLa cells. We provide protocols to detect ubiquitylated histones that are based on a combination of in vivo genetic and immunological assays. These methods should provide relatively simple and useful tools to study the global regulation of this important but poorly understood histone modification. PMID- 12893175 TI - Chromatin immunoprecipitation: a tool for studying histone acetylation and transcription factor binding. AB - The function of a protein in gene expression can often be explained, in part, by the location of that protein along a specific gene sequence. In recent years, the chromatin immunoprecipitation (ChIP) assay has been developed to study the association of proteins located within 2 A of DNA such as transcription factors and modified histones. Numerous important findings have been published using the ChIP assay and many questions about transcription have been answered. In this article, we present the ChIP assay currently used in our lab and discuss the various ways to optimize this assay for one's own use. PMID- 12893176 TI - Immunoprecipitation of native chromatin: NChIP. AB - Chromatin immunoprecipitation (ChIP) is widely used in many fields to analyze the distribution of specific proteins, or their modified isoforms, across defined DNA domains. ChIP procedures fall into two main categories, namely, those that use native chromatin prepared by nuclease digestion (designated NChIP), and those that use chromatin in which DNA and proteins are crosslinked, either chemically or with UV light (designated XChIP). Each procedure has its own advantages and drawbacks. Here, we outline the methods currently in use in our laboratory to isolate and immunoprecipitate native chromatin from cultured cells, and to isolate and analyze immunoprecipitated protein and DNA. PMID- 12893177 TI - Genomewide histone acetylation microarrays. AB - Histone acetylation and methylation are important regulators of gene activity. Chromatin immunoprecipitation (ChIP or ChrIP) has made it possible to examine not only the state of histone acetylation at a gene but also that of histone methylation and may soon be extended to other histone modifications such as phosphorylation and ubiquitination. In principle such studies are possible as long as an antibody is available to the particular histone modification. Once a target gene is identified it is instructive to see the effect of mutating putative enzymes responsible for the modification to determine how a particular enzyme is responsible for altering chromatin of that gene. Although specific target genes have been studied that contain such modifications recent technical advances have made it possible to study histone modifications genomewide. This not only allows for alternate views of particular paradigms to be investigated, but also uncovers chromosomal patterns of histone modification that would be missed in analyzing individual genes. We describe here an approach to rapidly study histone modifications genomewide by combining chromatin immunoprecipitation and DNA microarrays. PMID- 12893178 TI - In vivo protein-protein and protein-DNA crosslinking for genomewide binding microarray. AB - Chromatin immunoprecipitation (ChrIP or ChIP) has commonly been used to map protein-DNA interaction sites at specific genomic loci through use of formaldehyde-induced crosslinking. However, formaldehyde alone has proved inadequate for crosslinking of certain proteins such as the yeast histone deacetylase Rpd3. We report here a modified crosslinking procedure that includes a protein-protein crosslinking agent in addition to formaldehyde. Using this double crosslinking method, we have successfully mapped Rpd3 binding sites in vivo. We also describe the use of ChrIP in combination with DNA microarrays (ChrIP-array) to determine the pattern of Rpd3 binding genomewide. This approach couples the versatility of ChrIP with that of microarrays to identify binding patterns that would otherwise be hidden in a gene-by-gene survey. PMID- 12893179 TI - Histone deposition and chromatin assembly by RSF. AB - It is becoming clear that the structure of cellular chromatin is dynamic and capable of undergoing rapid changes to respond to the metabolic requirements of the cell. These changes have a direct impact on gene expression and, therefore, the chromatin context must be considered when biochemical reactions that involve DNA are studied. Over the past several decades, a number of methods for assembling chromatin in vitro have been described. Some of them use chemical compounds to deposit histone octamers onto the DNA. Others take advantage of cellular protein complexes that have the ability to assemble chromatin. Some of these complexes have been identified and purified. This article focuses on one of these factors, RSF (remodeling and spacing factor), which was identified in our laboratory. We describe how the chromatin assembly reaction is performed and how it can be monitored to evaluate its efficiency. PMID- 12893181 TI - Early ovarian cancer--time for a rethink on stage? PMID- 12893180 TI - Coupling tandem affinity purification and quantitative tyrosine iodination to determine subunit stoichiometry of protein complexes. AB - Rapid protein purification methodologies, such as strategies involving the tandem affinity purification module, have resulted in the identification of a tremendous number of multisubunit protein complexes. Furthermore, in this modern genomic age, mass spectrometry methods are often coupled with affinity purification to identify the genes that encode each protein subunit. However, simple methodologies to determine the stoichiometry of individual subunits within a multisubunit complex have not received much attention. In this article we describe a procedure to rapidly and efficiently determine the stoichiometry of subunits within multisubunit complexes using a combination of tandem affinity purification and quantitative 125I labeling of subunit tyrosines. PMID- 12893182 TI - MDM2 sensitizes a human ovarian cancer cell line. AB - OBJECTIVES: The purpose of this study was to determine whether overexpression of MDM2 could sensitize the ovarian cancer cell line A2780. METHODS: The wild-type p53-expressing cell line A2780 was stably transfected with pCMV-MDM2 (A2780-MDM2) or pCMV (A2780-V) as control. MTT assay and clonogenic survival assay were used to measure the cisplatin sensitivity. FACS and host cell (CAT) reactivation assay were used to estimate the change of cell cycle and ability of repairing cisplatin induced DNA damage. RESULTS: Parental A2780 and A2780-V had similar cisplatin sensitivities, whereas A2780-MDM2 was two- to threefold more sensitive to cisplatin. Repair of cisplatin-induced DNA damage was reduced in A2780 cells overexpressing MDM2, compared to A2780 cells in which wild-type p53 function was intact. After cisplatin treatment, A2780-MDM2 cells showed a pronounced S-phase arrest; however, A2780 cells with intact wild-type p53 arrested primarily in G2/M phase. CONCLUSIONS: MDM2 overexpression can increase cisplatin cytotoxicity in A2780, with loss of G1/S checkpoint control and decreased cisplatin-DNA adduct repair. This suggests that ovarian cancers that overexpress MDM2 may be amenable to treatment with platinum compounds. PMID- 12893183 TI - Technical modification of the nerve-sparing laparoscopy-assisted vaginal radical hysterectomy type 3 for better reproducibility of this procedure. AB - OBJECTIVE: The goal of this work is to make the technique of nerve-sparing laparoscopy-assisted vaginal radical hysterectomy (LAVRH) type 3 easier and less time consuming. METHODS: Compared with the original LAVRH type 3, laparoscopic preparation of the cardinal ligament is extended to the transection of the upper part of the rectovaginal ligament. In vaginal approach, the preparation begins not as originally with the bladder pillar and ureter, but with transection of the rest of the rectovaginal ligament followed by dorsal eversion of the uterus and, lastly, transection of the bladder pillar with preparation of the ureters. RESULTS: Thirty-two consecutive patients underwent the modified nerve-sparing LAVRH type 3. The mean operative time for the vaginal part of the procedure was 54.3 min, whereas for the original technique for LAVRH type 3, the mean operative time was 143.6 min. CONCLUSION: Prior transection of the rectovaginal ligament in the vaginal approach with dorsal eversion of the uterine fundus makes preparation of the ureter and transection of the bladder pillar easier and significantly less time consuming than in the original LAVRH type 3. PMID- 12893184 TI - AlphaV- and beta1-integrin subunits are commonly expressed in malignant effusions from ovarian carcinoma patients. AB - OBJECTIVE: The objective was to study expression of alphav- and beta1-integrin subunits in effusions, primary tumors, and solid metastases of ovarian carcinoma patients, as well as to evaluate its potential association with previously studied metastasis-associated molecules and clinicopathologic parameters. METHODS: Sections from 121 malignant effusions and 30 corresponding primary and metastatic lesions were evaluated for protein expression of the alphav- and beta1 integrin subunits using immunohistochemistry (IHC). A subset of effusions was additionally studied using immunoblotting (IB) and flow cytometry (FCM). mRNA in situ hybridization (ISH) was performed in 58 effusions and 30 biopsies. RESULTS: Protein expression of alphav- and beta1-integrin subunits was detected in carcinoma cells in 116/121 (96%) and 113/121 (93%) effusions, respectively. alphav protein expression was limited to carcinoma cells. IB and FCM confirmed IHC results. mRNA for alphav- and beta1-integrin subunits was detected in carcinoma cells in 37/58 (64%) and 33/58 (57%) effusions, respectively. Both protein and mRNA expression were higher in peritoneal effusions, significantly for alphav mRNA (P = 0.042) and beta1 protein (P = 0.023). beta1 protein expression in effusions was more frequently detected in better-differentiated tumors (P = 0.006). alphav-integrin subunit expression correlated with that of the previously studied matrix metalloproteinase-9 (MMP-9) (P = 0.006) and the MMP inducer EMMPRIN (P = 0.001). Expression of beta1-integrin subunit showed an association with that of EMMPRIN (P = 0.029), basic fibroblast growth factor (P < 0.001), and the MMP inhibitor TIMP-2 (P = 0.025). In carcinoma cells of solid lesions, alphav protein was uniformly present, while beta1 expression was limited to 15/30 (50%) specimens. As in effusions, protein expression of alphav subunit was cancer-specific, while beta1 protein was detected also in stromal fibroblasts and endothelial cells. CONCLUSIONS: The alphav- and beta1-integrin subunits are frequently expressed in ovarian carcinoma cells in effusions, and the alphav integrin subunit is a powerful diagnostic marker for carcinoma cells. The reduced expression of the beta1-integrin subunit in solid lesions may be attributed to the role of other subunits at these stages, such as the beta3 subunit as part of the alphavbeta3-vitronectin receptor. The high expression of integrin subunits with a role of binding mesothelium, invasion, and angiogenesis in carcinoma cells in both peritoneal and pleural effusions suggests that cells at both sites have metastatic potential. PMID- 12893185 TI - The detection of stage I ovarian cancer by three-dimensional sonography and power Doppler. AB - OBJECTIVE: The objective was to determine the diagnostic accuracy of three dimensional (3D) sonography and 3D power Doppler imaging, used together with standard 2D transvaginal grayscale and color/power Doppler modalities, for preoperative sonographic assessment of suspected ovarian lesions. METHODS: Five year retrospective analysis was performed by our experts on ultrasonography and surgery on the reports from 43 referred patients with suspected stage I ovarian cancer. All patients were evaluated during the week prior to surgery at our department. Preoperative sonographic assessment included careful examination of ovarian volume, morphology, and vascularity by four complementary sonographic methods. Scoring systems combining morphological and Doppler parameters were adopted for 2D and 3D sonographic examinations. Final diagnosis was confirmed by a histopathologist. RESULTS: Out of the 43 stage I ovarian cancers, 42 cases were successfully detected preoperatively by four complementary sonographic methods. Only 30 (69.8%) and 37 (86.1%) cases of stage I ovarian cancer were detected by 2D grayscale and combined 2D grayscale and color Doppler sonography, respectively. Morphological analysis obtained by 3D sonography alone detected 32 of 43 ovarian malignancies, reaching a diagnostic rate of 74.4%. Qualitative analysis of tumor vascularity architecture by 3D power Doppler significantly improved the sonographic management process and successfully detected 41 cases of stage I ovarian cancer (95.4%). When morphological features obtained by 3D sonography were added to 3D power Doppler findings, we achieved an even higher diagnostic accuracy of 97.7%. We found a statistically significant difference (P 0.50). However, the authors agreed upon the diagnosis of small cell carcinomas and undifferentiated carcinomas only in 2/3 of these high-risk diagnoses. Patients with high-risk diagnoses showed significantly lower overall survival than patients with non-high-risk diagnoses (P < 0.001). This inferior survival was independent of whether the reviewers had agreed on the high-risk diagnosis or not. CONCLUSION: Clinicians should be aware of the potential inconsistencies of histopathologic diagnoses. No histopathological classification system will ever be perfectly reproducible. Future histopathologic classification of the uterine cervix should emphasize the distinction between groups of particularly low or high prognostic risks. PMID- 12893189 TI - Laparoscopic detection of sentinel lymph nodes followed by lymph node dissection in patients with early stage cervical cancer. AB - OBJECTIVE: The purpose of this study was to investigate the feasibility of sentinel node detection through laparoscopy in patients with early cervical cancer. Furthermore, the results of laparoscopic pelvic lymph node dissection were studied, validated by subsequent laparotomy. METHODS: Twenty-five patients with early stage cervical cancer who planned to undergo a radical hysterectomy and pelvic lymph node dissection received an intracervical injection of technetium-99m colloidal albumin as well as blue dye. With a laparoscopic gamma probe and with visual detection of blue nodes, the sentinel nodes were identified and separately removed via laparoscopy. If frozen sections of the sentinel nodes were negative, a laparoscopic pelvic lymph node dissection, followed by radical hysterectomy via laparotomy, was performed. If the sentinel nodes showed malignant cells on frozen section, only a laparoscopic lymph node dissection was performed. RESULTS: One or more sentinel nodes could be detected via laparoscopy in 25/25 patients (100%). A sentinel node was found bilaterally in 22/25 patients (88%). Histological positive nodes were detected in 10/25 patients (40%). One patient (11%) had two false negative sentinel nodes in the obturator fossa, whereas a positive lymph node was found in the parametrium removed together with the primary tumor. In seven patients (28%), the planned laparotomy and radical hysterectomy were abandoned because of a positive sentinel node. Bulky lymph nodes were removed through laparotomy in one patient, and in six patients only laparoscopic lymph node dissection and transposition of the ovaries were performed. These patients were treated with chemoradiation. In two patients, a micrometastasis in the sentinel node was demonstrated after surgery. Ninety-two percent of all lymph nodes was retrieved via laparoscopy, confirmed by laparotomy. Detection and removal of the sentinel nodes took 55 +/- 17 min. Together with the complete pelvic lymph node dissection, the procedure lasted 200 +/- 53 min. CONCLUSION: Laparoscopic removal of sentinel nodes in cervical cancer is a feasible technique. If radical hysterectomy is aborted in the case of positive lymph nodes, sentinel node detection via laparoscopy, followed by laparoscopic lymph node dissection, prevents potentially harmful and unnecessary surgery. PMID- 12893190 TI - Three-dimensional coculture of endometrial cancer cells and fibroblasts in human placenta derived collagen sponges and expression matrix metalloproteinases in these cells. AB - OBJECTIVE: Collagen gel constitutes a valuable tool for the study of cell-matrix interactions; however, it is sometimes difficult to use the gel, in which tumor and stromal cells are cocultured, for immunohistochemistry, because it is easily broken during the process of fixation and embedding in paraffin, especially after long-term culture. METHODS: To examine the interaction between endometrial cancer cells and fibroblasts in tumor invasion, we carried out three-dimensional (3-D) coculture of various endometrial cancer cell lines and fibroblasts in human placenta derived collagen sponges and analyzed the expression and localization of matrix metalloproteinases (MMP) and plasminogen activators (PA) in these cells by immunohistochemistry. RESULTS: After 4 weeks of culture on the collagen sponges, endometrial cancer cells composed stratiform or glandular structures on the layer of extracellular matrix, which was composed from fibloblasts and extracellular matrix. Compared to Ishikawa cells, which were rarely invasive, HEC-1A and HEC 1BE and cocultured fibroblasts showed high invasiveness and strong expression of some proteins. In cell line HEC-1BE, MMP-1, -7, and -9, MT1-MMP, tissue inhibitors of metalloproteinases 2, and uPA showed intensive staining in both cancer cells and fibroblasts by immunohistochemistry. HEC-1A cells and cocultured fibroblasts showed expression patterns similar to those of HEC-1BE. CONCLUSION: These results suggested that expression of MMPs and uPA was accelerated in fibroblasts surrounded by cancer cells. We believe that our 3-D coculture system has merit in that the interaction between cancer cells and stromal cells can be visually analyzed by immunohistochemistry and that experiments for a long period, at least 2 weeks, are possible. Furthermore, it is expected that some animal, e.g., nude mouse, experiments can be replaced by experiments using this culture system. PMID- 12893191 TI - Frozen section examination of the endocervical margin of cervical conization specimens. AB - OBJECTIVE: We conducted this retrospective study to determine accuracy of frozen section examination of endocervical margin during cold knife conization. METHODS: Between June 1993 and June 2001, 310 consecutive patients underwent cervical conization for squamous intraepithelial lesion or stage IA1 cervical cancer. Before 1997, the surgical specimens of 149 patients were processed following a standard pathological procedure (historical group). After 1997, a frozen section of the upper endocervical margin was processed during surgery for 161 patients. If the upper endocervical margin was involved with intraepithelial neoplasia, the surgeon performed a second resection if possible. Results of the frozen section examination were compared with the final diagnoses to determine sensitivity, specificity, and positive and negative predictive values. The usefulness of this procedure was evaluated by comparison of positive margin status rate with the one of the historical control group. RESULTS: For the diagnosis of intraepithelial neoplasia involving the endocervical margin, the sensitivity, specificity, and positive and negative predictive values of frozen section were 91%, 100%, 100%, and 98%, respectively. Eleven patients had definitive positive endocervical margin in the frozen section group (three false negatives, six patients without additional resection, and two patients with intraepithelial neoplasia involving the upper margin of the additional resection) and 17 patients in the historical group (P =.16). CONCLUSION: Frozen section examination of the endocervical margin of cervical specimen obtained during cold knife conization is highly accurate. Its clinical relevance has to be demonstrated in a multicenter study. PMID- 12893192 TI - Association between high-risk human papillomavirus DNA load and precursor lesions of cervical cancer in Mexican women. AB - OBJECTIVE: Our objective was to determine the association between viral load of high risk human papilloma virus (HPV) using the Hybrid Capture II (HC II) system and cervical intraepithelial neoplasia (CIN) lesion stage. METHODS: A total of 182 consecutive women with confirmed diagnoses of CIN 1-3 and 182 healthy women with negative Pap were included. All subjects underwent structured interviews focused on socioeconomic and reproductive factors. HC II testing was used to detect human papilloma virus (HPV) DNA. Viral load was measured by light measurements expressed as relative lights unit (RLU) ratio (specimens/control). Log(10)RLU ratios were categorized for analysis into four groups: negative (50% carcinoma cells (n = 21, 20%), >50% sarcoma cells (n = 13, 12%), or sarcoma cells only (n = 2, 2%). The microscopic analysis demonstrated a preponderance of epithelial cells in the primary setting and suggested the epithelial component to drive the tumor, a finding consistent with the monoclonal theory. Biopsies derived from surgery for recurrent disease (n = 8) consisted of carcinoma cells only (0%), >50% carcinoma cells (n = 1, 13%), >50% sarcoma cells (n = 4, 50%), or sarcoma cells only (37%). Since sarcomatous cells dominated the tumorigenic cell population in the recurrent setting, this analysis revealed a change of the composition of metastatic lesions in time when compared to the data in the primary setting. This change was supported by the observation of a threefold higher incidence of sarcoma-dominated metastatic lesions at interval debulking when compared to primary debulking (24 vs 8%, respectively). The potential of a phenotypic change during ovarian cancer progression was further highlighted by the detection of two cases of carcinosarcoma that presented as a recurrence of epithelial ovarian carcinoma. CONCLUSION: Our results are consistent with the monoclonal theory of ovarian carcinosarcoma histogenesis, but suggest that there is a tendency toward a sarcomatous differentiation during disease progression. These data are important to understand the tumor biology and might have implications for a tailored treatment of ovarian carcinosarcoma. PMID- 12893203 TI - Role of KRAS and BRAF gene mutations in mucinous ovarian carcinoma. AB - OBJECTIVES: The goals of this study were to perform a comprehensive assessment of the prevalence of KRAS oncogene mutations in invasive epithelial ovarian carcinomas of various histologic subtypes, and for any subgroup(s) in which KRAS mutation was found to be common, to address the hypothesis that those tumors without KRAS mutation had sustained alternative activation of this signaling pathway through mutation of the BRAF oncogene. METHODS: A total of 104 primary, invasive epithelial ovarian carcinomas from a 10-year period at this institution were selected for study based on histologic classification. The histologic cell type was serous in 21 cases, endometrioid in 30 cases, clear cell in 31 cases, and mucinous in 22 cases. Additional clinical and pathological information was abstracted from patient records, and pathology review was performed for all cases. Direct sequence analysis of exon 2 of the KRAS gene, containing codon 12, was performed using DNA isolated from all tumor specimens. Sequence analyses of exons 11 and 15 of the BRAF gene were performed for the 22 cases of mucinous ovarian carcinoma. RESULTS: Activating KRAS mutations were more common in mucinous tumors (50%) than in all other histologic types combined (5%; P < 10( 7)). Mutation of KRAS was more common in stage I tumors than in advanced stage tumors (P = 0.0004). Of the 11 mucinous tumors with KRAS mutations, 6 were of Mullerian (endocervical) type and 5 were of gastrointestinal type. No mucinous tumor was found to harbor a BRAF mutation. CONCLUSIONS: These data indicate that KRAS oncogene mutations exist in several histologic types of invasive epithelial ovarian carcinoma, especially stage I tumors, but are common only in tumors of mucinous histology. Mutations are equally prevalent in mucinous ovarian cancers of Mullerian and gastrointestinal types. In contrast to other solid tumor types frequently affected by KRAS mutation, mucinous ovarian cancers without a KRAS mutation have not sustained alternative activation of this signaling pathway through mutation of the BRAF oncogene. PMID- 12893204 TI - Recurrent endometrial polyps in postmenopausal breast cancer patients on tamoxifen. AB - OBJECTIVES: Endometrial polyps are the most common endometrial pathology described in association with postmenopausal tamoxifen exposure, with an incidence of up to 10.7% of malignancy. Some women tend to develop recurrent polyps. However, no one has yet described any risk factors for the development of recurrent endometrial polyps in postmenopausal breast cancer tamoxifen-treated patients. METHODS: We compared various clinical features of 64 postmenopausal breast cancer tamoxifen-treated patients with a primary endometrial polyp (Group I), with those of 27 similar patients with recurrent polyps (Group 2). RESULTS: Previous exposure to hormone replacement therapy was significantly more common and duration of tamoxifen treatment, up to the diagnosis of primary endometrial polyp, was significantly shorter in Group II patients (P = 0.0217 and P = 0.0148, respectively). Logistic regression analysis revealed that the combination of shorter tamoxifen exposure before the diagnosis of primary polyp, lower parity, lower menopausal age at the diagnosis of primary polyp, and higher years of tamoxifen treatment was found to increase significantly the risk of developing recurrent endometrial polyps. Any additional year of tamoxifen treatment may increase by fivefold the risk of developing recurrent polyps. There was no significant difference in ultrasonographic endometrial thickness measured before resection of the primary polyps in both groups and before the resection of recurrent polyps in Group II. CONCLUSIONS: Previous use of HRT, shorter duration of tamoxifen exposure, and additional years of tamoxifen treatment may significantly increase the risk of developing recurrent endometrial polyps in postmenopausal breast cancer tamoxifen-treated patients. PMID- 12893205 TI - Stage Ia1 cervical squamous cell carcinoma: conservative management after laser conization with positive margins. AB - OBJECTIVE: Cone margin status has been reported to be the most important predictor of recurrent disease in patients with cervical intraepithelial neoplasia (CIN) undergoing cervical conization. Our purpose was to evaluate the conservative management of selected patients with microinvasive (FIGO stage Ia1) squamous cell carcinoma who have been treated by cervical conization with positive margins. METHODS: Twenty-seven patients underwent KTP laser conization and vaporization for stage Ia1 squamous cell carcinoma followed by careful observation. Involved margins were diagnosed if CIN III or more was present at the ectocervical or endocervical margin and 7 patients formed the basis of the present study. Follow-up consisted of cytology, histology, and pelvic examination. Disease recurrence was defined as a histology diagnosis of CIN III or more on colposcopically directed biopsy or endocervical curettage. RESULTS: The endocervical margins were involved by carcinoma in situ in seven (26%) patients. No ectocervical margin involvement was detected. No lymph-vascular space involvement (LVSI) and confluent invasion were seen. All seven patients were free of recurrent disease during median follow-up of 4.0 (range 2.3-7.6) years. CONCLUSION: These results suggest that laser conization and vaporization may be a reasonable treatment option in patients with microinvasive (FIGO Stage Ia1) squamous cell carcinoma despite positive cone margins without invasive disease when LVSI is not demonstrated. PMID- 12893206 TI - Relative influences of tumor volume before surgery and the cytoreductive outcome on survival for patients with advanced ovarian cancer: a prospective study. AB - OBJECTIVE: The purpose of this study was to determine the relative influences of the extent of disease present before surgery and completeness of cytoreduction on survival for patients with advanced ovarian cancer. METHODS: Patients (408) with stage IIIC epithelial ovarian cancer had cytoreductive surgery before systemic platinum-based combination chemotherapy. A ranking system (0-3) was devised to prospectively quantify the extent of disease involving: (1) right upper quadrant (diaphragm/hepatic, and adjacent peritoneal surfaces), (2) left upper quadrant (omentum/gastro-colic ligament, spleen, stomach, transverse colon, splenic flexure of colon), (3) pelvis (reproductive organs, recto-sigmoid, pelvic peritoneum), (4) retroperitoneum (pelvic/aortic nodes), and (5) central abdomen (small bowel, ascending/descending colon, mesentery, anterior abdominal wall, pericolic gutters). Survival was analyzed (log rank and Cox regression) on the basis of the rankings at these anatomic regions, the sum of intraabdominal rankings, and the cytoreductive outcome. RESULTS: Overall median and estimated 5 year survivals were 58.2 months and 49%. On univariate analysis, the central abdominal (P = 0.008) and left upper quadrant (P = 0.03) rankings, the sum of rankings (P = 0.01), and the cytoreductive outcome (P 1 cm residual, RR 2.98; P = 0.001). CONCLUSIONS: Cytoreduction to a visibly disease-free outcome has a more significant influence on survival than the extent of metastatic disease present before surgery. Operative efforts should not be abbreviated on the hypothesis that extensive disease at specific anatomic regions precludes long-term survival. PMID- 12893207 TI - Breast cancer metastatic to abdomen and pelvis: role of surgical resection. AB - OBJECTIVE: The purpose of this study was to describe the characteristics and outcome of women with metastatic breast cancer to the abdomen and pelvis, and to assess the role of surgical resection of abdominal and pelvic metastasis in this disease. METHODS: We retrospectively reviewed the medical records of 59 women with documented metastatic breast cancer to the abdomen or pelvis who had exploratory surgery by the Gynecology Service between 1986 and 2001. RESULTS: Exploratory surgery was performed a median of 5 years (range, 0-25 years) after initial diagnosis of breast cancer. Median survival from diagnosis of abdominal disease was 23 months, and 5-year survival was 24%. Survival was 36 months for optimally debulked patients (<2 cm of residual disease) and 20 months for suboptimally debulked patients (P = 0.07). Patients diagnosed 5 or more years after initial breast cancer diagnosis had a median survival of 36 months versus 17 months if diagnosed earlier (P < 0.01). On multivariate analysis the time to recurrence of breast cancer in the abdomen and optimal debulking were both significant variables. Hazard ratio for dying of disease if recurring before 5 years was 2.7 (CI 1.45-5.03) [P < 0.01]. Hazard ratio for dying of disease if suboptimal debulking was achieved was 2.14 (CI 1.13-4.02) [P = 0.02]. CONCLUSIONS: The disease pattern of metastatic breast carcinoma to the abdomen and pelvis does not appear to effect survival. Survival in patients where optimal debulking is achieved and in those recurring late is improved. Surgical resection of metachronous metastatic breast cancer to the abdomen and pelvis may be an important component of the management of this disease and should be considered in candidate patients. PMID- 12893208 TI - The proto-oncogene c-kit is expressed in leiomyosarcomas of the uterus. AB - OBJECTIVE: The proto-oncogene c-kit encodes for a 145-kDa transmembrane tyrosine kinase receptor. Interaction with its ligand, stem cell factor, is essential in the development of hematopoietic stem cells, mast cells, gametocytes, melanocytes, and interstitial cells of Cajal. C-kit expression has been identified in a number of different neoplasms that includes mastocytosis/mast cell leukemia, acute myeloblastic leukemia, seminoma/dysgerminoma, and gastrointestinal stromal tumors. This study examines c-kit expression in uterine endometrial stromal sarcomas, leiomyomas, and leiomyosarcomas using immunohistochemistry. METHODS: Archival tissue from 38 patients with the uterine mesenchymal tumors (16 leiomyosarcomas, 8 leiomyomas, 11 low-grade endometrial stromal sarcomas, and 3 high-grade endometrial stromal sarcomas) was stained with polyclonal antibody for c-kit. Modified avidin biotin (ABC) immunoperoxidase method was employed for antibody detection. Individual tumors were considered positive if more than 10% of the cells comprising the neoplasm displayed immunoreactive staining. Staining intensity was graded 1+ to 3+ and distribution graded as focal (10-30% of the cells), intermediate (30-60% of the cells), or diffuse (>60% of the cells). RESULTS: C-kit was positive in 12 (75%) of the 16 leiomyosarcomas. The staining was 3+ and diffuse in the majority of the positive tumors. C-kit expression was not detected in any of the 8 leiomyomas. Two of the 3 high-grade endometrial stromal sarcomas displayed c-kit positivity. Staining was diffuse and 3+ in both of these tumors. Expression of c-kit was observed in only 3 of the 11 low-grade endometrial stromal sarcomas. CONCLUSIONS: C-kit is expressed in uterine leiomyosarcomas and endometrial stromal sarcomas. Adjunctive diagnostic studies using c-kit may be useful in distinguishing leiomyosarcomas from benign leiomyomas in uterine tumors that offer uncharacteristic features. Furthermore, studies should investigate the prospect of treating these malignant tumors with tyrosine kinase inhibitors. PMID- 12893209 TI - Surgery as sole treatment for serous borderline tumors of the ovary with noninvasive implants. AB - OBJECTIVES: The objectives were to describe the clinical characteristics and prognosis of surgically treated patients with stage II and III serous borderline tumors of the ovary with noninvasive implants. MATERIALS AND METHODS: From 1990 to 2000, 16 patients with stage II and III ovarian serous borderline tumors and noninvasive implants were diagnosed and prospectively followed at our center. All patients underwent surgical treatment including staging and their pathology was reviewed. Fifteen patients had thorough surgical staging by laparotomy, while one patient was staged laparoscopically. No patient was treated with adjuvant therapy (radiation or chemotherapy) after surgical treatment and none were lost to follow up. RESULTS: The mean age at diagnosis was 42 years (range 26-59). Fourteen patients were treated by abdominal hysterectomy, bilateral salpingo-oophorectomy, omentectomy, and multiple peritoneal biopsies, while 2 patients were treated conservatively for fertility preservation. Two patients underwent pelvic and para aortic lymph node dissection. Fifteen of 16 patients had ovarian surface involvement with tumor. All patients but 2 had clinical evidence of extraovarian disease at the time of surgery. The mean duration of follow-up was 60.7 months (range 2-134 months). Thirteen patients (81%) are alive without evidence of disease. Four patients (25%) required subsequent surgery for recurrent disease and all are still alive. Two patients have been treated with chemotherapy (paclitaxel/carboplatin) for progressive borderline disease, while an additional patient was treated after first relapse with chemotherapy for an invasive recurrence. CONCLUSIONS: Carefully staged patients with advanced serous borderline tumors of the ovary and noninvasive implants have a good prognosis without adjuvant therapy. PMID- 12893210 TI - Enhancement of sensitivity to cisplatin by orobol is associated with increased mitochondrial cytochrome c release in human ovarian carcinoma cells. AB - OBJECTIVES: Based on our previous report showing that orobol, a potent phosphatidylinositol 4-kinase (PI4K) inhibitor, produced cisplatin (DDP) sensitivity, we have determined the mechanism of orobol-sensitization effect. METHODS AND RESULTS: Orobol produced >2-fold DDP sensitivity in human ovarian carcinoma 2008 cells and its DDP-resistant variant 2008/C13*5.25 cells (C13). Because orobol had no effect on conventional mechanisms such as DDP accumulation or cellular metallothionein and glutathione content, we have focused on the apoptotic signaling pathway. Orobol induced a significant increase in apoptosis in DDP-treated cells, as estimated by frequency of condensed nuclear chromatin with Hoechst 33342 stain, although orobol alone did not have any effect on apoptotic potential. The caspase-3-inhibiting peptide Ac-DEVD-CHO completely inhibited the orobol sensitization effect but did not block DDP cell cytotoxicity per se. Orobol rendered both of these cells resistant to rhodamine 123 (Rh) by more than 2.5-fold, indicating significant decrease of mitochondrial membrane potential (DeltaPsim). Confocal laser microscopy of cells stained with the mitochondria (MT)-specific dye Rh revealed that orobol decreased Rh-fluorescent intensity. Electron microscopy of these cells showed that orobol induced swelling and condensation of MT. Orobol suppressed both naturally expressed and the DDP induced Bcl-2 expression significantly. Orobol and DDP treatment reduced cytochrome c level in MT determined by Western blot analysis, indicating increased amount of cytochrome c release from MT, whereas orobol alone did not alter the amount of cytochrome c in MT. CONCLUSIONS: These results indicate that orobol produced DDP sensitivity in human ovarian carcinoma cells by inducing apoptosis through the MT-dependent signaling pathway. PMID- 12893212 TI - Slide review in gynecologic oncology ensures completeness of reporting and diagnostic accuracy. AB - OBJECTIVE: We reviewed patient records in our tertiary care teaching hospital to assess the value of the mandatory slide review policy in gynecologic oncology with emphasis on completeness of reports. METHODS: Cases reviewed between October 2001 to September 2002 were studied. Clinical information was gathered from discussions at the weekly tumor board and from chart review. The standardized reporting guidelines in benchmark surgical pathology textbooks were used to assess the completeness of original pathology reports of excisional specimens. Diagnostic discrepancies were classified as major if the resultant change led to alteration of management or minor if it did not. RESULTS: Three hundred fifty-one cases were reviewed; 173 biopsies and 178 excisional specimens. Only 140 (78.7%) of the original pathology reports of the latter group conformed to standardized reporting guidelines. Of the 38 incomplete reports, 18 were missing critical information necessary for planning of further therapy, representing 10.1% of reports of all excisional specimens. We agreed with the original diagnosis in 252 cases (71.8%). Minor discrepancies were noted in 70 (19.9%) and major discrepancies in 29 cases (8.3%). No major discrepancy resulted from reviewing any of the vulvar specimens or cases that were already reviewed by gynecologic pathologists of other academic institutes. CONCLUSION: Mandatory slide review in gynecologic oncology is an important component in the management of gynecologic cancer patients because it completes reporting on missing parameters required for planning subsequent therapy in 10.1% of cases and recognizes discrepancies altering management in 8.3% of patients. PMID- 12893211 TI - A feasibility study on biweekly administration of docetaxel for patients with recurrent ovarian cancer. AB - OBJECTIVE: A recent study demonstrated that docetaxel (DTX) was an effective agent for second line chemotherapy against ovarian cancer. Weekly administration of taxane compounds had been more effective compared with a 3 week interval administration in ovarian cancer. The role of biweekly administration of DTX has been unknown. We conducted a dose determination and feasibility study of biweekly DTX administration in patients with ovarian cancer. METHODS: Patients with histologically confirmed epithelial ovarian cancer who received one or more regimens of prior chemotherapy with more than 4 weeks of treatment-free interval were eligible. DTX was administered as 1-h intravenous infusion every two weeks for at least four courses. The starting dose was 40 mg/m(2) (level 1) and the dose was escalated to 50 mg/m(2) (level 2) and 60 mg/m(2) (level 3) in consequent patient cohorts. RESULTS: Nine patients were examined in this study. The treatments were completely performed in all cohorts. Mean treatment delay ranged from 0 to 2.0 days. Dose level did not affect treatment delay. At the first dose level, no patients experienced grade 3/4 neutropenia. Two patients in level 2 experienced grade 3/4 neutropenia. In level 3, all patients had grade 4 neutropenia. Nonhematologic toxicities were tolerable. Of eight patients with measurable disease, all patients in level 1 showed progressive disease, and all patients in level 2 were no-change. There were two responders showing complete response and partial response and one case was no-change in level 3. CONCLUSION: The present study showed that biweekly administration of 60 mg/m(2) DTX was feasible for recurrent ovarian cancer. PMID- 12893213 TI - The effects of CO2 pneumoperitoneum on the survival of women with persistent metastatic ovarian cancer. AB - OBJECTIVES: The effects of CO(2) pneumoperitoneum on the survival of women with metastatic ovarian cancer have not been documented. We sought to describe the survival of women with persistent stage III-IV ovarian cancer as documented by positive second-look laparoscopy or laparotomy and to see whether the laparoscopic approach with CO(2) pneumoperitoneum has a negative effect on overall survival. METHODS: We conducted a retrospective review of all patients with FIGO stage III-IV invasive epithelial ovarian cancer who were found to have persistent disease at second-look surgery. All patients underwent primary surgery followed by intravenous chemotherapy and were clinically without evidence of disease prior to second-look surgery. Second-look laparoscopy began to be utilized regularly in 1994. The selection of the second-look surgical approach depended on the surgeon's discretion. CO(2) pneumoperitoneum was utilized for all laparoscopic cases with the maximum intra-abdominal pressure maintained at 15 mm Hg. Patients received a variety of additional intravenous, intraperitoneal, or oral chemotherapy following positive second-look surgery. RESULTS: Between 6/1/91 and 6/30/02, 289 patients were found to have persistent ovarian/peritoneal cancer at second look. Second-look operations included 131 (45%) transperitoneal laparoscopies and 139 (48%) laparotomies. Nineteen (7%) patients underwent laparoscopy followed immediately by laparotomy. The mean age, stage distribution, histology, grade, and size of residual disease at second look did not differ between the two groups. The median overall survival for patients who underwent laparoscopy, 41.1 months (95% CI, 33.2-58.1), did not significantly differ from that of the laparotomy group, 38.8 months (95% CI, 31.9-44.2) (P = 0.742). CONCLUSIONS: Transperitoneal laparoscopy with CO(2) pneumoperitoneum does not appear to reduce the overall survival of women with persistent metastatic intra abdominal carcinoma of ovarian/peritoneal origin. The overall survival appears to be independent of the second-look surgical approach. PMID- 12893214 TI - Urinary matrix metalloproteinases as a potential screening test for gynecologic malignancies. AB - OBJECTIVE: This was a pilot study to determine the feasibility of using urinary matrix metalloproteinases (MMPs) as a screening test for gynecologic malignancies. METHODS: Urine samples from ovarian (n = 29), cervical (n = 31), endometrial (n = 31), and vulvar (n = 5) cancer patients and 19 controls were collected. Substrate gel electrophoresis (zymography) was used to determine the presence of MMP-2 (72 kDa), MMP-9 (92 kDa) and two high-molecular-weight forms (130 and 220 kDa) of MMPs. The sensitivity, specificity, and positive and negative predictive values of the test for each tumor type were determined. RESULTS: No association was noted between malignancy and presence of urinary MMPs in ovarian, cervical, endometrial, or vulvar cancer patients. Sensitivity, specificity, negative and positive predictive values, and likelihood ratios were determined. Sensitivities ranged from 28.1 to 51.0% for individual MMPs, peaking at 69.8% when the presence of any of the four proteinases was considered a positive test. Specificities ranged from 42.1 to 68.4%. CONCLUSION: Our study suggests that the presence of MMPs in the urine of patients with a gynecologic malignancy is not an adequate screening test for disease. There was also little evidence of an association between urinary MMPs and stage or extent of disease. However, the limited number of patients in the various stages of each disease site, specifically advanced stage disease, make it difficult to state this definitively. PMID- 12893215 TI - Microinvasive squamous cell carcinoma of the cervix: immunohistochemically detected prognostic factors in a case with poor clinical outcome. AB - BACKGROUND: Established prognostic factors for cervical cancer are tumor stage, vascular space invasion, tumor size/volume, depth of invasion, and lymph node status. Although patients with superficially invasive lesions have a very good prognosis, cases are still reported which have a poor clinical outcome. CASE: We report a case of a 41-year-old woman with very early stromal invasion (0.6 mm deep and <1 mm wide) who presented with a pelvic recurrence at 3 years and then developed an anterior abdominal wall and disseminated peritoneal recurrence 4 years after extrafascial hysterectomy. Archival tissue of the cone biopsy was stained immunohistochemically for CD44v6, Her2-neu, p53, bcl-2, MMP-1, and VEGF and showed positive staining for CD44v6 and MMP-1. CONCLUSION: The identification of new prognostic factors may enhance our understanding of the biologic behavior of early invasive cancer of the cervix. Our findings suggest that CD44v6 and MMP 1 may be markers worth further investigation in patients with microinvasive cervical cancer. PMID- 12893216 TI - Abdominal wall metastasis of a cervical adenocarcinoma at the laparoscopic trocar insertion site after ovarian transposition: case report and review of the literature. AB - BACKGROUND: Laparoscopic ovarian transposition is currently used in the preirradiation management of cervical cancer in young women. This surgical technique helps avoid the short- and long-term complications of early menopause. Because there remains a risk of metastasis at the site where the laparoscopic trocar is inserted, more precise indications for this surgery are required. CASE: We report the case of a patient with an abdominal wall metastasis that followed laparoscopic ovarian transposition performed before radiation therapy and surgical treatment for a stage IIb cervical adenocarcinoma. Observations during the laparotomy nonetheless led us to resect the transposed ovary during the laparotomy. The pathology examination of the ovary found a microscopic metastasis. Several months later, a left abdominal parietal nodule, corresponding to a metastasis of the adenocarcinoma, was found on the site through which the trocar had been inserted during the laparoscopy and was probably associated with the ovarian transposition. CONCLUSION: This is the only published case to describe an abdominal wall metastasis due to laparoscopic trocar insertion during ovarian transposition. The decision to perform a laparoscopic ovarian transposition in cervical cancer must take into account the frequency of trocar site metastases, which seems to be higher with advanced-stage cervical cancers, in cases of lymph node involvement and especially in adenocarcinomas. PMID- 12893217 TI - Leiomyosarcoma of the broad ligament: a case report and literature review. AB - Primary leiomyosarcoma of the broad ligament is a very rare, rapidly progressive and highly malignant gynaecological tumor. There are only 11 cases reported so far in the literature. An 87-year-old Caucasian lady with leiomyosarcoma of the broad ligament is presented. Treatment consisted of resection and total abdominal hysterectomy with bilateral salpingo-oopphorectomy. Unusually, she developed large recurrent pelvic disease in the absence of disseminated metastasis following surgery. She died within 8 weeks of primary surgery. This highlights the unknown behavior of this tumor and the need for more research into management. PMID- 12893218 TI - Fused functional-anatomic images of metastatic cancer of cervix obtained by a combined gamma camera and an X-ray tube hybrid system with an illustrative case and review of the 18F-fluorodeoxyglucose literature. AB - BACKGROUND: 18F-Fluorodeoxyglucose (FDG) assessments have provided clinically important information in cervical cancer. FDG studies can now be performed by both dedicated PET systems and by new-generation gamma cameras. Hybrid systems which consist of positron emission tomography (PET) or a gamma camera with X-ray for fusion of functional-anatomic data without changing the patient's position are now available. CASE: A woman with newly diagnosed cervical cancer underwent preoperative FDG studies using a hybrid gamma camera. In addition to the known primary tumor, FDG detected heretofore unidentified metastatic disease at the liver, bone, and para-aortic lymph nodes: the treatment approach was consequently altered to chemo- and radiotherapy. The fused images provided precise localization of the lesions, guiding bone biopsy and radiation field planning. CONCLUSION: When PET is unavailable, a modified gamma camera can provide clinically relevant data in patients with cervical cancer. PMID- 12893219 TI - Activity of chemotherapy with carboplatin plus paclitaxel in a recurrent mesonephric adenocarcinoma of the uterine corpus. AB - BACKGROUND: Malignant lesions derived from mesonephric (Wolffian) remnants are uncommon. The course of these tumors is usually indolent, and the recurrence has only been documented in nine cases. Because of the small number of cases, no current recommendations exist regarding treatment, and little is known about the response to chemotherapeutic agents. CASE: A 33-year-old woman was diagnosed with a mesonephric adenocarcinoma arising in the uterine corpus. Ten months after initial surgery and radiotherapy she presented with local and pulmonary relapse. Salvage chemotherapy with carboplatin plus paclitaxel was administered with a good response. CONCLUSIONS: Mesonephric adenocarcinomas are uncommon neoplasms. Their treatment remains elusive. We report a case of a recurrent uterine mesonephric adenocarcinoma that presented a good response to therapy with carboplatin plus paclitaxel. A review of the previous literature is also presented. PMID- 12893220 TI - A case of ovarian cancer with remote metastases, with emphasis on changes in tumor marker values. AB - BACKGROUND: The prognosis of cancer patients with distant metastases is extremely poor. CASE: A 51-year-old Japanese woman with Stage IIIc serous cystoadenocarcinoma of the ovary achieved complete remission. At 30 and 39 months after the initiation of therapy, solitary brain and then spleen metastases without intraperitoneal lesions were found, respectively. A simultaneous elevation in tumor markers (CA125 and CA72-4) was noted at the spleen metastasis; however, only a marked elevation in CA 72-4 levels at the brain metastasis was noted. Immunohistochemical studies revealed that although CA125-positive cells could be observed in both the original tumor and the spleen metastasis, CA125 positive cells could not be observed in the brain metastasis and that CA72-4 positive cells could be observed in both the original ovarian tumor and the metastatic tumor. CONCLUSION: It is important to monitor with tumor markers with different characteristics, such as CA125 and CA72-4. PMID- 12893221 TI - Radical trachelectomy via a trans-sacral approach: a case report. AB - BACKGROUND: Endometrial cancer may arise in patients previously treated for rectal cancer. Surgical management can be challenging, and the gynecologic oncologist may need to employ alternative surgical approaches. CASE: A 71-year old female with a history of Dukes class B rectal cancer previously treated by abdominoperineal resection and pelvic radiation therapy subsequently developed an endometrial cancer. Initial surgical management was difficult, and the entire cervix could not be removed. The patient recurred locally the following year and was managed by radical trachelectomy via a trans-sacral approach. CONCLUSION: Locally recurrent endometrial cancer can pose difficult management decisions. Radical resection may be the only alternative. Unusual situations may require adaptation of different surgical approaches of which the gynecologic oncologist must be aware. PMID- 12893222 TI - Symptomatic skeletal muscle metastasis and elephantiastic lymphedema in a patient with recurrent ovarian carcinoma. AB - BACKGROUND: Distant tumor metastases in ovarian cancer patients in striated muscles are extremely rare. So far, only one case of metastatic but asymptomatic spread to striated muscle has been described. CASE: A patient with recurrent ovarian carcinoma presented with a gluteal mass 1 week following optimal debulking surgery. CT and MRI initially suggested muscle necrosis or intramuscular bleeding. Increasing pain and massive lymphedema led to immobilization and clinical deterioration. Repeated ultrasound and CT-guided biopsy confirmed the diagnosis of skeletal muscle metastases. CONCLUSION: This is the first case description of distant skeletal muscle metastasis as the leading clinical symptom of late stage ovarian carcinoma. It reveals possible limitations of CT and MRI as diagnostic tools for early identification of striated muscle metastases. PMID- 12893223 TI - A case of lymphoma occurring in an ovarian teratoma. AB - BACKGROUND: We describe the discovery, by pathology, of a totally unexpected diagnosis involving an ovarian teratoma. CASE: A middle-aged woman underwent routine total abdominal hysterectomy and bilateral salpingoophrectomy for ultrasonically diagnosed uterine fibroids and a coexistent benign right ovarian cyst. The operative findings were of fibroid uterus and right ovarian teratoma. However, pathology revealed a malignant lymphoma occurring in, and arising from, the teratoma. CONCLUSION: The routine removal of a presumed ovarian teratoma may yield unexpected results. Ovarian teratomas are indeed cellular chimeras. PMID- 12893224 TI - A case report of multicentric verrucous carcinoma of the female genital tract. AB - BACKGROUND: Verrucous carcinoma is a variant of squamous cell carcinoma with distinct features including slow locally invasive growth and verrucous appearance. Verrucous carcinoma of the vagina is considered an extremely rare lesion because only 17 cases have been reported in the literature. CASE: We report a case of vaginal verrucous carcinoma with a second focus in the cervix. The patient was treated with surgery and adjuvant interferon therapy for local recurrence. Human papillomavirus was detected in both vaginal and cervical tumor tissue by immunohistochemistry. CONCLUSION: Diagnosis of verrucous carcinoma may be difficult, particularly if biopsy specimen involves only the surface epithelium. The role of human papillomavirus as an etiologic agent in verrucous carcinoma is still a matter of discussion. Effective management requires surgical resection. The efficiencies of radiotherapy and interferon therapy are discussed. PMID- 12893225 TI - Gangliocytic paraganglioma arising from mature cystic teratoma of the ovary. AB - BACKGROUND: Gangliocytic paraganglioma is a rare neoplasm involving the small intestine, stomach, and spinal cord. Ovarian gangliocytic paraganglioma has not been reported in the medical literature. CASE: A 55-year-old caucasian woman underwent exploratory laparotomy, total abdominal hysterectomy, and bilateral salpingo-oophorectomy for evaluation of a persistent right adnexal mass. Microscopic examination of the right ovary revealed a mature cystic teratoma with a mural nodule consistent with gangliocytic paraganglioma. As there was no evidence of significant pleomorphism or neoplastic infiltration, surgical staging was not performed. CONCLUSION: Gangliocytic paraganglioma may arise from ovarian cystic teratoma. Although most cases of gangliocytic paragangliomas are benign, surgical staging and retroperitoneal lymphadenectomy may be required if histopathology is suggestive of invasive disease or if enlarged lymph nodes are noted. PMID- 12893226 TI - Mullerian adenosarcoma of vagina arising in persistent endometriosis: report of a case and review of the literature. AB - BACKGROUND: Primary adenosarcoma arising in vaginal endometriosis poses a diagnostic challenge, especially in superficial vaginal biopsies. CASE: A 56-year old woman, with a prior diagnosis of ovarian endometriosis, presented with a rapidly enlarging mass of the vaginal vault. Two prior biopsies were benign and showed endometriosis. The third vaginal biopsy revealed benign endometriotic glands cuffed by a cellular stroma with moderate cytologic atypia, a histological appearance diagnostic of Mullerian adenosarcoma. A 16-cm vaginal mass that had infiltrated the pelvic structures was resected. CONCLUSIONS: Close clinical follow-up of extrauterine endometriosis and clinical-pathologic correlation is necessary. Histological features such as cellular stromal cuffing around benign endometriotic glands are critical in arriving at a timely diagnosis of adenosarcoma in patients with persistent extrauterine endometriosis, even in superficial vaginal biopsies. PMID- 12893227 TI - BRCA1/2-related ovarian cancers are of tubal origin: a hypothesis. PMID- 12893228 TI - Importance of the general practitioner in the early detection of ovarian cancer. PMID- 12893229 TI - Sentinel lymph node procedure in early vulvar cancer. PMID- 12893230 TI - Vesicocutaneous fistula mimicking an abdominal wall abscess 2 years after radical abdominal hysterectomy. PMID- 12893231 TI - Vaginal cuff recurrence of endometrial cancer treated by laparoscopic-assisted vaginal hysterectomy. PMID- 12893234 TI - Flynn et al.: does the interval from primary surgery to chemotherapy influence progression-free survival in ovarian cancer? PMID- 12893236 TI - Concern with "Is there a survival benefit to adjuvant radiotherapy in high-risk surgical stage I endometrial cancer?". PMID- 12893238 TI - ISG15, not just another ubiquitin-like protein. AB - ISG15 is a ubiquitin-like protein containing two ubiquitin homology domains and becomes conjugated to a variety of proteins when cells are treated with type I interferon or lipopolysaccharide. Although ISG15 shares several common properties with those of other ubiquitin-like molecules, it is a unique member, whose expression and conjugation to target proteins are tightly regulated by specific signaling pathways, indicating it may be associated with specialized functions in innate immune system. Loss of UBP43 (USP18), a protease that specifically removes ISG15 from ISG15-modified proteins, in mice leads to decreased life span, brain cell injury, and hypersensitivity to interferon stimulation. In UBP43 deficient cells, interferon induces a prolonged Stat1 tyrosine phosphorylation and DNA binding, which result in a prolonged and enhanced activation of interferon stimulated genes. PMID- 12893239 TI - Directional surface plasmon-coupled emission: A new method for high sensitivity detection. AB - Fluorescence emission is nearly isotropic in space. With typical optical components the collection efficiency is 1% or less. In this preliminary report, we describe a novel approach to transforming the normally isotropic emission into directional emission with a collection efficiency near 50%. This can be accomplished for fluorophores located near a semi-transparent silver film on a glass substrate. The emission couples with the surface plasmon resonance on the silver surface and enters the transparent substrate at a sharply defined angle, the surface plasmon angle for the emission wavelength. We estimate that 40-70% of the total emission enters the substrate at the plasmon angle and can thus be directed towards a detector. Background emission from fluorophores distant from the silver does not couple with the plasmon and is not detected. Different emission wavelengths couple at different angles allowing spectral discrimination without additional optics. Surface plasmon-coupled emission represents a new technology which can be used for high detection efficiency with microfluidic and/or surface-bound assay formats. PMID- 12893240 TI - Effects of leptin gene expression in mice in vivo by electroporation and hydrodynamics-based gene delivery. AB - In vivo electroporation and hydrodynamics-based gene delivery were utilized to test the effect of leptin gene transfer on food intake, and body and fat weights of mice. Gene transfer of pVRmob by electroporation caused a significant reduction in body weight compared with the control counterpart (p<0.05), although a lesser effect was found in food intake, and the weights of interscapular brown and epididymal fat by electroporation. As might be expected, the hydrodynamics based transfection method significantly reduced body weight over 1 week post transfection (p<0.05). Furthermore, epididymal fat was decreased by 50% at 1 week after gene transfer (p<0.001). These results suggest that both electroporation and hydrodynamics-based gene delivery may be effective approaches for systemic delivery of recombinant leptin to the central nervous system, and that the efficiency of gene transfer in hydrodynamics-based gene delivery was markedly higher than that in electroporation at least within the first week after transfection. PMID- 12893241 TI - Deletion mutants of protective antigen that inhibit anthrax toxin both in vitro and in vivo. AB - The anthrax toxin complex is primarily responsible for most of the symptoms of anthrax. This complex is composed of three proteins, anthrax protective antigen, anthrax edema factor, and anthrax lethal factor. The three proteins act in binary combination of protective antigen plus edema factor (edema toxin) and protective antigen plus lethal factor (lethal toxin) that paralyze the host defenses and eventually kill the host. Both edema factor and lethal factor are intracellularly acting proteins that require protective antigen for their delivery into the host cell. In this study, we show that deletion of certain residues of protective antigen results in variants of protective antigen that inhibit the action of anthrax toxin both in vitro and in vivo. These mutants protected mice against both lethal toxin and edema toxin challenge, even when injected at a 1:8 ratio relative to the wild-type protein. Thus, these mutant proteins are promising candidates that may be used to neutralize the action of anthrax toxin. PMID- 12893242 TI - Lipopolysaccharide down-regulates inducible nitric oxide synthase expression in swine heart in vivo. AB - Studies of the regulation of iNOS expression have provided many contradictory results. Comparing iNOS expression profile between cell types or organs of the same animal under the same experimental conditions may provide an explanation for these conflicting results. We have examined iNOS mRNA and protein expression in heart and liver of the same group of pigs. We found that there is a sharp difference in iNOS expression between heart and liver. The iNOS mRNA and protein was constitutively expressed in the heart at high level, but was not detectable in the liver of the same control animal. Lipopolysaccharide (LPS, 100 microg/kg, i.v.) caused a marked iNOS induction in the liver, but significantly down regulated iNOS expression in the heart. This differential iNOS expression appears to be physiologically relevant, since LPS and the iNOS inhibitor, S methylisothiourea, exerted different effects on hepatic and myocardial blood flow. Our data demonstrate a fundamental difference in iNOS regulation in the heart and liver of swine, and may explain the contradictory data on the regulation of iNOS expression. PMID- 12893243 TI - Centaurin-alpha(1) associates with and is phosphorylated by isoforms of protein kinase C. AB - Centaurin-alpha(1) is a member of the family of ADP-ribosylation factors (ARF) GTPase activating proteins (GAPs), although ARF GAP activity has not yet been demonstrated. The human homologue, centaurin-alpha(1) functionally complements the ARF GAP activity of Gcs1 in yeast. Although Gcs1 is involved in the formation of actin filaments in vivo, the function of centaurin remains elusive. We have identified a number of novel centaurin-alpha(1) binding partners; including CKIalpha and nucleolin. In this report, we have focused on the interaction of centaurin-alpha(1) with PKC. All groups of PKC associate directly through their cysteine rich domains. Centaurin-alpha(1) is also a substrate for all PKC classes and we have identified the two sites of phosphorylation. This is the first report of a kinase that phosphorylates centaurin-alpha(1). PMID- 12893244 TI - Insulators to improve expression of a 3(')IgH LCR-driven reporter gene in transgenic mouse models. AB - A locus control region (LCR) containing four transcriptional enhancers lies downstream of the IgH chain locus. We studied transgenes carrying a 3(')IgH LCR driven GFP reporter gene for expression and B cell differentiation stage specificity. We also compared transgenes that were or were not flanked by two copies of the beta-globin HS4 insulator, an element defined by its ability to protect transgenes from the influences of surrounding genes at the insertion site. Results indicate that insulators are instrumental in sustaining GFP expression in GFP-3(')LCR transgenic mice when they were included. Flow cytometry experiments reported a strictly B cell specific GFP expression from pre-B cells in bone marrow to mature B cells in spleen. Despite addition of 5(')HS4 insulators to the GFP-3(')LCR construct, complete transgene silencing occurred in some transgenic lines and was systematically observed in ageing animals from all lines. PMID- 12893245 TI - Maize tonoplast PP(i)-dependent H(+)/Ca(2+) exchange: two K(s) for Ca(2+) and inhibition by thapsigargin. AB - Maize root tonoplasts are able to accumulate Ca(2+) using the energy derived from the H(+) gradient formed during PP(i) hydrolysis. Oxalate increases 6- to 10-fold the amount of Ca(2+) accumulated by tonoplast. Two apparently different K(s) values for Ca(2+) with values of 0.36 and 4.70 microM were detected when oxalate was included in the medium and the free Ca(2+) concentration in the medium was buffered with the use of EGTA. Binding of Ca(2+) to the outer surface of tonoplasts inhibits the outflow of Ca(2+) previously accumulated by the tonoplast, half-maximal inhibition being observed in presence of 1 microM Ca(2+). Thapsigargin, a specific inhibitor of Ca(2+)-ATPase, inhibits the Ca(2+) uptake driven by H(+) gradient but does not inhibit the hydrolysis of PP(i) nor the formation of a H(+) gradient. PMID- 12893246 TI - Anoxia-reoxygenation-induced cytochrome c and cardiolipin release from rat brain mitochondria. AB - Rat brain mitochondria were successively submitted to anoxia and reoxygenation. The main mitochondrial functions were assessed at different reoxygenation times. Although the respiratory control ratio decreased, the activity for each one of the enzymes participating in the respiratory chain was not affected. However, during reoxygenation, mitochondrial membrane lipoperoxidation quickly increased and was proportional to the decrease seen in membrane fluidity. Under the same conditions, cytochrome c and cardiolipin were released from mitochondria and their rate of release increased with reoxygenation time. The release of cytochrome c and cardiolipin was followed by the collapse of the membrane potential and it was not inhibited by cyclosporin A. Addition of the antioxidant alpha-tocopherol abolished all these reoxygenation-induced changes. These data indicate that, in this model, reoxygenation promotes the uncoupling of respiratory chain, and cytochrome c and cardiolipin releases. These events are not related to the membrane potential collapse but to an oxidative stress. PMID- 12893247 TI - Baculoviral p35 inhibits oxidant-induced activation of mitochondrial apoptotic pathway. AB - In this study we report that the baculovirus p35 anti-apoptotic protein prevents cell death by quenching free radicals at a very upstream step in the apoptotic pathway. Mitochondria of activated rat peritoneal macrophages as well as Spodoptera frugiperda (Sf9) insect cells, following treatment with oxidants, H(2)O(2)/UVB irradiation, release cytochrome c followed by activation of caspase 3. Transfection of macrophages/Sf9 cells with a construct carrying the p35 gene under the CMV/HSP promoters resulted in p35 expression and consequent arrest of oxidative stress-induced apoptosis. p35 expression also inhibited cytochrome c release from the mitochondria of oxidant-exposed cells and blocked caspase-3 activation. PMID- 12893248 TI - Expression and promoter analysis of mouse mastrin gene. AB - Human astrin is a newly identified microtubule-associated protein, which is highly expressed in the testis. Silencing of astrin has resulted in growth arrest and apoptotic cell death. In this study, we describe the cloning and genomic structure of mastrin, the mouse counterpart to astrin. The overall mouse mastrin amino-acid sequence is 66% identical to human astrin. Mastrin protein was demonstrated to localize to mitotic spindles during mitosis. Genomic clones containing mastrin gene were isolated; the gene was found to have 24 exons spanning 24kb of genomic DNA. Deletion analysis of 5(')-flanking sequences demonstrated that the first 120bp proximal to the TATA-less promoter region is necessary for minimal transcription of the mouse mastrin gene. PMID- 12893249 TI - Epitope mapping of gibberellin to the anti-gibberellin A(4) monoclonal antibody by saturation transfer difference NMR spectroscopy. AB - Saturation transfer difference (STD) NMR spectroscopy is a promising tool for rapid screening, identifying ligands that interact with a target protein, and characterizing the epitopes of the ligands. Gibberellins (GAs) are a class of plant hormones and form a large family consisting of more than 120 members. A few of them, called "active" GAs, are considered to be perceptible to a receptor that remains unknown. We applied STD NMR spectroscopy to detect the binding activity and identify the binding epitope of gibberellin A(3) (GA(3)) that is recognized by monoclonal antibody 4-B8(8)/E9. This is one of the antibodies that can mimic a GA receptor in the manner of recognition of active GAs. The information on the binding epitope, obtained by STD NMR, was in good agreement with that shown by analyzing the crystal structure of the antibody-GA(4) complex. This suggests that STD NMR spectroscopy would be very useful to characterize the interaction between GAs and such binding proteins as GA-catabolic enzymes and receptors. PMID- 12893250 TI - Tissue- and stressor-specific differential expression of two hsc70 genes in carp. AB - Two genes expressing 70 kDa heat shock proteins were identified in Cyprinus carpio. The sequence similarities and the intron-interrupted structure of the coding regions indicate that carp Hsc70-1 and Hsc70-2 belong to the Hsp70 cognate subfamily. The expressions of the two hsc70 genes were followed by semi quantitative RT-PCR. Both genes are expressed under unstressed conditions in a characteristic tissue-specific manner. Inducibility of the response to elevated temperature, cold shock, and Cd treatment was investigated in the liver and muscle, in whole-animal experiments. Both genes were insensitive to or only weakly induced by the stressors, with two exceptions: Cd treatment resulted in an 11-13-fold enhanced induction of hsc70-1 in the liver and cold shock enhanced induction of hsc70-2 in the muscle by 7.5-10-fold. PMID- 12893251 TI - Fanconi anemia cell lines deficient in alphaII spectrin express normal levels of alphaII spectrin mRNA. AB - Fanconi anemia is a genetic disorder characterized by hypersensitivity to DNA interstrand cross-linking agents and a defect in the ability to repair this type of damage. This deficiency correlates with reduced levels of alphaII spectrin, a structural protein involved in the repair of DNA interstrand cross-links. The present study addresses the question of whether the reduced levels of alphaII spectrin in FA-A, FA-C, and FA-G cells are due to reduced expression of this protein and/or due to differences in the three regions of alternate splicing of alphaII spectrin mRNA. Relative quantitative RT-PCR showed that levels of alphaII spectrin mRNA in the three FA cell lines were similar to normal as were the sites of alternative mRNA splicing. These results indicate that decreased levels of alphaII spectrin in these FA cell lines are not due to reduced expression of alphaII spectrin mRNA or due to differences in regions of alternate splicing of these transcripts, but rather appear to be related to reduced stability of alphaII spectrin in these cell lines. PMID- 12893252 TI - Scoring and identifying organism-specific functional patterns and putative phosphorylation sites in protein sequences using mutual information. AB - The co-variance of amino acid positions within a multiple alignment of 294 protein kinases from mammals, plants, and bacteria was studied. Applying mutual information (MI), characteristic amino acid sites have been identified markedly discriminating the different organisms. The relation of surface accessibility of these sites in the 3D structure of a kinase and their MI content is studied. We extended the method to score a predicted phosphorylation site of this highly conserved catalytic protein kinase region. Based on this score mammalian and plant protein kinases were grouped together apart from the bacterial kinases. Thus, the presented method allows us to analyse putative phosphorylation sites in the context of their organism-specific origin. PMID- 12893253 TI - Identification and characterization of the murine TRPM4 channel. AB - The transient receptor potential (TRP) channels form a superfamily with six transmembrane structures, which is common in other types of voltage-dependent channels. The TRP-melastatin (TRPM) subfamily includes the putative tumor suppressor melastatin, which was originally found as a down-regulated protein in melanoma tumor cell lines. Here, we report a novel TRP-related protein that is a murine orthologue of human TRPM4. The function of the novel murine TRPM4 was studied in HEK-293 cells using a fluorescent calcium indicator, fura-2. The removal and re-introduction of extracellular calcium triggered changes in the intracellular calcium only in cells expressing TRPM4a, which suggests that this novel channel plays a role in the calcium entry process. We also isolated a splice variant of TRPM4 that was proven to be non-functional. Both TRPM4 variants integrated into the plasma membrane. Furthermore, FRET analysis revealed that TRPM4a and TRPM4b localized close together, suggesting a multimerization of the two molecules. PMID- 12893254 TI - Structural determinants in the stability of the serpin/proteinase complex. AB - Serpins inhibit serine proteinases through formation of stable 1:1 complexes. In this study we have evaluated the effects of PAI-1 neutralizing antibodies (MA) on the stability of PAI-1/proteinase complexes, partially destabilized through prolongation of the reactive center loop. MA-8H9D4, reacting with residues Arg(300), Gln(303), and Asp(305), had no effect on the stability. In contrast, MA 33H1F7 and MA-55F4C12, reacting with alpha-helix F and the turn connecting hF with s3A, affected significantly and proteinase-dependently formed PAI 1/proteinase complexes. That is, MA-33H1F7 increased the stability of both PAI 1/t-PA and u-PA complexes (7- and 3-fold, respectively) whereas MA-55F4C12 stabilized PAI-1/t-PA complexes (3-fold) but destabilized PAI-1/u-PA complexes (2 fold). It is concluded that interference with the docking site of the cognate proteinase in the preformed serpin/proteinase complex may affect the intrinsic stability. We hypothesize that this is the consequence of a decreased or increased torsion of the RCL on the catalytic triad in the proteinase. PMID- 12893255 TI - A novel lectin from the wild mushroom Polyporus adusta. AB - A lectin with antiproliferative activity toward tumor cell lines and mitogenic activity toward splenocytes was isolated from the mushroom Polyporus adusta. The lectin was composed of two identical subunits each with a molecular weight of 12 kDa. It was adsorbed on both DEAE-cellulose and Q-Sepharose and unadsorbed on CM Sepharose. The hemagglutinating activity of the lectin was inhibited by turanose and by a large variety of other carbohydrates. It was adversely affected in the presence of NaOH or HCl at a concentration of 7.5mM and above, and when the ambient temperature was raised above 70 degrees C. All divalent and trivalent metallic chlorides tested at 1.25-10mM including CaCl(2), MgCl(2), ZnCl(2), MnCl(2), and AlCl(3), did not alter the hemagglutinating activity of the lectin. FeCl(3) at 10mM caused the hemagglutinating activity to increase by 100%, but it did not change the lectin activity when tested at lower concentrations up to 5mM. PMID- 12893256 TI - Differences in the expression of lipolytic-related genes in rat white adipose tissues. AB - We have investigated in vivo whether the gene expression of the beta3-adrenergic receptor (beta3-AR), perilipin A, hormone-sensitive lipase (HSL), and adipocyte lipid-binding protein (ALBP/aP2) is regulated in a site-specific manner. To induce lipolysis and discriminate between short- and long-term modifications, rats were submitted to an experimental fast for one or five days followed or not by refeeding. The mRNA encoding beta3-AR in retroperitoneal adipose tissue (RP) was significantly increased by one and five days of fasting (4-fold) and then lowered by one day of refeeding (2-fold) compared to fed rats. The reverse trend was observed for perilipin A expression in one day fasted rats. HSL mRNA concentrations were significantly induced (2.2-fold) by five days of fasting relative to fed animals and remained high after refeeding. ALBP/aP2, peroxisome proliferator-activated receptor gamma, and CAAT/enhancer binding protein alpha mRNA levels were essentially unaffected by dietary manipulations. Fasting and/or refeeding were similarly ineffective at regulating gene expression in SC. These data provide a molecular basis for regional differences at different steps of the lipolytic process. PMID- 12893257 TI - Oxidation of glyceraldehyde-3-phosphate dehydrogenase enhances its binding to nucleic acids. AB - Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a protein with various activities far from its enzymatic function. Here, we showed that the oxidation of SH-groups of the active site of GAPDH enhanced its binding with total transfer RNA or with total DNA. Both NAD and NADH-the cofactors of GAPDH-inhibited the GAPDH-RNA (DNA) interaction, though NAD was much less effective than NADH in the case of oxidized GAPDH. Oxidation of GAPDH strongly decreased its affinity to NAD but not to NADH. Immobilized tetramers of GAPDH dissociated into dimers during the incubation with total RNA but not DNA. The staining of HeLa cells with monoclonal antibodies specific to dimers, monomers or the denatured form of GAPDH revealed the condensation of non-native forms of GAPDH in the nucleus. The role of oxidation of GAPDH in the regulation of the quaternary structure of the enzyme and in its interaction with nucleic acids is discussed. PMID- 12893258 TI - Irreversibility of chilling injury in zucchini squash (Cucurbita pepo L) could be a programmed event long before the visible symptoms are evident. AB - Zucchini fruits were subjected to 2.5 or 10 degrees C for 16d, followed by transfer to 20 degrees C for 24h in order to evaluate the relationship between ripening pattern, measured as CO(2) evolution and ethylene (C(2)H(4)) production, and metabolic heat production (q). Chilling injury (CI) visible symptoms were evident after 8d at 2.5 degrees C, but none were recorded on fruits kept at 10 degrees C. In fruits held at 10 degrees C, q, C(2)H(4) production, and CO(2) evolution diminished in the course of 16d, whereas in those at 2.5 degrees C CO(2) evolution showed an early burst peaking at 8d. Both C(2)H(4) production and q also showed a burst at 2.5 degrees C but they started at 4 and 8d, respectively, and peaked at 12d. The results showed that irreversibility of chilling injury in zucchini could occur long before the appearance of visible symptoms, although the metabolic activity accompanying the irreversibility process was not noticeable by isothermal calorimetry. PMID- 12893259 TI - Differential influence of proteolysis by calpain 2 and Lp82 on in vitro precipitation of mouse lens crystallins. AB - The purpose of the present study was to compare the susceptibility of crystallins proteolyzed by ubiquitous calpain 2 and by lens-specific calpain Lp82 to insolubilization. To test this, transgenic (TG) mice expressing a calpain 2, in which the active site cysteine 105 was mutated to alanine, were produced. Expression of mutated calpain 2 was driven in lens by coupling the mutated gene to the betaB1-crystallin promoter. Light scattering was measured in solutions of lens proteins after activation of endogenous calpain 2 and/or Lp82. Mass spectrometric analysis was performed to determine the cleavage sites and the calpain responsible for insolubilization of crystallins. Lens proteins from TG mice incubated in vitro with calcium showed higher light scattering compared to proteins from wild type (WT) mice. alphaA-crystallin from TG mice was proteolyzed by Lp82. In contrast, alphaA-crystallin in lenses from WT mice were proteolyzed by both calpain 2 and Lp82. These results suggested that Lp82-induced proteolysis of crystallins caused increased susceptibility of truncated crystallins to in vitro precipitation. Since Lp82 is highest in young animals, Lp82-induced proteolysis and precipitation may be one of the factors responsible for the cataract formation in young rodents. PMID- 12893260 TI - Arabinosylcytosine downregulates thymidine kinase and induces cross-resistance to zidovudine in T-lymphoid cells. AB - The aim of this study was to determine molecular mechanism(s) responsible for the reduced thymidine kinase activity (TK) observed earlier in an arabinosylcytosine (araC) resistant lymphoid cell line (H9-araC cells), which was obtained following continuous cultivation of H9 cells in the presence of 0.5 microM araC. Compared to H9 cells, in H9-araC cells TK1 and TK2 gene expressions were reduced to 17.7% and 2.5%, respectively, and the cellular AZT accumulation was diminished to 35.8%. These cells were also found cross-resistant to azidothymidine (>42-fold). There was no significant difference in the expression of MDR1, MRP4 or TK protein. The lack of correlation between the expressions of TK protein and TK1 and TK2 suggests that post-translational factors may also play a role in the reduced TK activity in H9-araC cells. These findings suggest that araC affects TK expression at the genetic level. PMID- 12893261 TI - Molecular characterization of a novel nucleolar protein, pNO40. AB - We report the discovery and characterization of a novel nucleolar protein. This protein, referred to as pNO40 based on its molecular weight on SDS-PAGE, was identified through yeast two hybrid interaction screen of a human kidney cDNA library using pinin (pnn) protein as the bait. The deduced amino acids of pNO40 derived from cDNA cloning of diverse species display high conservation; 95% identify between human and mouse and 57.3% identity for human and zebrafish. Several distinct domains are discernable in the ORF of pNO40, including a ribosomal protein S1 RNA binding region, a CCHC type zinc finger, and clusters of basic amino acid representing potential nucleolar targeting signal. Immunostaining of endogenous or transfected pNO40 indicated that it is localized to nucleoli of diverse cultured cells, with some concentration in the granular component of nucleoli. Northern blot analysis demonstrated that pNO40 message is expressed ubiquitously across all tissues examined. Characterization of human and mouse pNO40 gene revealed that mouse gene spans 44 kb in length and contains 8 exons, while that of human is 68 kb in length and displays two isoforms generated by alternative splicing of the 5(')-untranslated region and differential usage of translation start site. Based on sequence features and its subcellular location, we predict that pNO40 is a novel nucleolar protein with function related to ribosome maturation and/or biogenesis. PMID- 12893262 TI - Mitochondrial ribosomes in a trypanosome. AB - The nature, and even the existence, of trypanosome mitochondrial ribosomes has been the subject of some debate. We investigated this further in the insect trypanosome, Crithidia fasciculata. In sucrose gradients of parasite lysates, mitochondrial ribosomal RNA co-sediments at approximately 35S with nascent peptides synthesized in the presence of the cytosolic translational inhibitor, cycloheximide. Co-sedimenting peptides in this peak are much reduced when the parasites are treated with the bacterial translational inhibitor, chloramphenicol. In CsCl gradients this peak resolves at a buoyant density of 1.42 g/cm(3), a value typical for mito-ribosomes. Electron microscopy of peak material shows particles smaller than cytosolic ribosomes, but with characteristic ribosomal shapes. We propose that these particles represent the parasite's mitochondrial ribosomes. PMID- 12893263 TI - Expression of recombinant Clostridium difficile toxin A using the Bacillus megaterium system. AB - Pathogenic Clostridium difficile produces two major protein toxins, toxin A and toxin B. We used the Bacillus megaterium expression system for expression of recombinant toxin A. The construct for the toxin A gene was obtained by the following cloning strategy: the gene for toxin A was generated in three parts, each of them ligated into a cloning vector. The three parts were sequentially fused to the complete gene. The holotoxin gene was ligated into the expression vector pWH1520. This vector was modified to generate a toxin with a C-terminally located His-tag. Gene expression in the B. megaterium system resulted in an approximate 300 kDa protein, which was identified by specific antibody as toxin A. Recombinant, His-tagged toxin A was purified by Ni(2+) as well as thyroglobulin affinity chromatography. Characterization of the recombinant toxin A showed identical cytotoxicity and in vitro-glucosyltransferase activity as the native toxin A from C. difficile. PMID- 12893265 TI - Molecular cloning and expression analysis of the aryl hydrocarbon receptor of Xenopus laevis. AB - The aryl hydrocarbon receptor (AHR) is a member of the basic helix-loop-helix/Per Arnt-Sim (bHLH/PAS) family of transcription factors. Although this receptor has been known to mediate the toxic effects of environmental pollutants, its physiological functions remain elusive. Here, we describe the isolation and expression pattern of the Xenopus AHR gene. The predicted amino acid sequence contained regions characteristic of other vertebrate AHRs. However, in line with previously described fish AHR genes, no distinct Q-rich domain was found. Phylogenetic analysis demonstrated that Xenopus AHR was clustered within the AHR1 clade. As in the case of mammalian AHR genes, the Xenopus AHR gene was expressed in all the adult tissues tested. Xenopus AHR was also expressed during early development, in parallel with expression of the CYP1A7 gene, which is thought to be regulated by AHR. These results suggest that while frogs are relatively tolerant to TCDD toxicity, the AHR of frogs has characteristics similar to those of other vertebrate AHRs. PMID- 12893264 TI - A sphingosine-dependent protein kinase that specifically phosphorylates 14-3-3 (SDK1) is identified as the kinase domain of PKCdelta: a preliminary note. AB - A specific protein kinase that phosphorylates Ser60, Ser59, or Ser58 of 14-3 3beta, eta, or zeta, respectively, only in the presence of sphingosine (Sph) or N,N-dimethyl-Sph (DMS), was termed "sphingosine-dependent protein kinase-1" (SDK1) [J. Biol. Chem. 273(34) (1998) 21834]. We have now identified SDK1 as a protein having the same amino acid sequence as in the C-terminal-half kinase domain of PKCdelta, with approximately 40 kDa molecular mass, based on large scale purification of a protein from rat liver, and partial sequence using three different combinations of LC-MS or LC-MS/MS with respective search engine. PKCdelta did not display any SDK1 activity and PKCdelta activity was inhibited by Sph and DMS. However, strong SDK1 activity, only in the presence of Sph or DMS, became detectable when PKCdelta was incubated with caspase-3, which releases the approximately 40 kDa kinase domain. PMID- 12893266 TI - The solution structure of the oxidized bovine microsomal cytochrome b(5) mutant V61H. AB - Using 1488 NOE constraints, 19 stereo-specific assignments, 13 pairs of H-bond constraints, and 140 pseudo-contact shift constraints, a family of 35 structures of bovine microsomal cytochrome b(5) mutant V61H has been obtained through the program PSEUDYANA. The family has been further refined by restrained energy minimization to give a family of final structures. The RMSD values of final structures with respect to the average structure are 0.45+/-0.11 and 0.96+/-0.10A for backbone and heavy atoms, respectively. The final Deltachi(ax) and Deltachi(rh) values are 2.34 x 10(-32) and -0.67 x 10(-32)m(3), respectively. The comparisons between the solution structures of mutant V61H and WT cytochrome b(5), and X-ray structure of the mutant V61H show that the global folding of the molecule in solution is unchanged and the side-chain of His61 deviates from the heme pocket and extends into the solvent like in its crystal structure. However, the helices around the heme pocket undergo outward global displacement while their local conformations are well maintained. Meanwhile, the heme ring shows a little off the heme pocket, which accounts for the lower stability of the mutant. Additionally, the axial ligand rings counterclockwise rotate around His39 N-Fe axis due to the mutation, which is confirmed by variation of the hyperfine shifts of the heme protons of V61H compared to those of WT cytochrome b(5). PMID- 12893267 TI - Cytochrome p450 complement (CYPome) of the avermectin-producer Streptomyces avermitilis and comparison to that of Streptomyces coelicolor A3(2). AB - The genus Streptomyces produces about two-thirds of naturally occurring antibiotics and a wide array of other secondary metabolites, including antihelminthic agents, antitumor agents, antifungal agents, and herbicides. The newly completed genome sequence of the avermectin-producing bacterium Streptomyces avermitilis contains 33 cytochromes p450 (CYPs), many more than the 18 observed in Streptomyces coelicolor A3(2). Some of the likely metabolic functions are reported together with their genomic location and bioinformatic analysis. Seven entirely new CYP families were found together with close homologues of some forms observed in S. coelicolor A3(2). The presence of unusual CYP forms associated with conservons is revealed and of these, CYP157 forms in both S. avermitilis and S. coelicolor A3(2) deviate from the previously accepted rule for an EXXR motif within the K-helix of CYPs. Amongst this range of CYPs are forms associated with avermectin, filipin, geosmin, and pentalenolactone biosynthesis as well as unknown pathways of secondary metabolism. PMID- 12893268 TI - A constitutively expressed 36 kDa exochitinase from Bacillus thuringiensis HD-1. AB - A 36 kDa chitinase was purified by ion exchange and gel filtration chromatography from the culture supernatant of Bacillus thuringiensis HD-1. The chitinase production was independent of the presence of chitin in the growth medium and was produced even in the presence of glucose. The purified chitinase was active at acidic pH, had an optimal activity at pH 6.5, and showed maximum activity at 65 degrees C. Of the various substrates, the enzyme catalyzed the hydrolysis of the disaccharide 4-MU(GlnAc)(2) most efficiently and was therefore classified as an exochitinase. The sequence of the tryptic peptides showed extensive homology with Bacillus cereus 36 kDa exochitinase. The 1083 bp open reading frame encoding 36 kDa chitinase was amplified with primers based on the gene sequence of B. cereus 36 kDa exochitinase. The deduced amino-acid sequence showed that the protein contained an N-terminal signal peptide and consisted of a single catalytic domain. The two conserved signature sequences characteristic of family 18 chitinases were mapped at positions 105-109 and 138-145 of Chi36. The recombinant chitinase was expressed in a catalytically active form in Escherichia coli in the vector pQE-32. The expressed 36 kDa chitinase potentiated the insecticidal effect of the vegetative insecticidal protein (Vip) when used against neonate larvae of Spodoptera litura. PMID- 12893269 TI - Purification and characterization of a novel glutathione S-transferase from Atactodea striata. AB - A novel GST isoenzyme was purified from hepatopancreas cytosol of Atactodea striata with a combination of affinity chromatography and reverse-phase HPLC. The molecular weight of the enzyme was determined to be 24 kDa by SDS-PAGE electrophoresis and 48 kDa by gel chromatography, in combination with GST information from literature revealed that the native enzyme was homodimeric with a subunit of M(r) 24 kDa. The purified enzyme, exhibited high activity towards 1 chloro-2,4-dinitrobenzene (CDNB) and 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole (NBD Cl). Kinetic analysis with respect to CDNB as substrate revealed a K(m) of 0.43 mM and V(max) of 0.24 micromol/min/mg and a specific activity of 108.9 micromol/min/mg. The isoelectric point of the enzyme was 5.5 by isoelectric focusing and its optimum temperature was 38 degrees C and the enzyme had a maximum activity at approximately pH 8.0. The amino acid composition was also determined for the purified enzyme. PMID- 12893270 TI - Bone morphogenetic protein signalling in NGF-stimulated PC12 cells. AB - Bone morphogenetic proteins (BMPs) are shown to potentiate NGF-induced neuronal differentiation in PC12 phaeochromocytoma cells grown on collagen under low-serum conditions. Whereas, cell bodies remained rounded in control medium or with only BMPs present, addition of BMP4 or BMP6 robustly increased the neuritogenic effect of NGF within 2 days. NGF-increased phosphorylation of p44(Erk1) and p42(Erk2) between 2 and 24h was unaffected by addition of BMP6. PC12 cells transfected with the SBE(4x)-luc reporter showed that BMP4 significantly increased receptor activated Smad activity. Expression of constitutively active BMP receptor ALK2 activating Smad1 and Smad5 resulted in a strong increase in the SBE(4x)-luc reporter response. Adding the inhibitory Smad7 drastically reduced this signal. In contrast to wild-type (wt) Smad5, a Smad5 variant lacking five Erk phosphorylation sites in the linker region (designated Smad5/5SA) showed a strong background transcriptional activity. A fusion construct (Gal4-Smad5/5SA) was also highly transcriptionally active. Addition of the MEK inhibitor U0126 to PC12 cells expressing Gal4-Smad5/wt did not increase background transcriptional activity. However, upon activation by constitutively active ALK2 both Gal4 Smad5/wt and Gal4-Smad5/5SA strongly stimulated transcription. The data show that serine residues of the linker region of Smad5 reduce spontaneous transcriptional activity and that NGF-activated Erk does not antagonise BMP signalling at this site. Hence, NGF and BMP signals are likely to interact further downstream at the transcriptional level in neuronal differentiation of the PC12 cells. PMID- 12893271 TI - CCR5 N-terminus peptides enhance X4 HIV-1 infection by CXCR4 up-regulation. AB - The HIV-1 envelope glycoprotein gp120 interacts consecutively with CD4 and CCR5 to mediate the entry of R5-HIV-1 strains into target cells. The N-terminus of CCR5, which contains several sulfated tyrosines, plays a critical role in gp120 CCR5 binding and, consequently, in viral entry. Here, we demonstrate that a tyrosine sulfated peptide, reproducing the entire N-terminal extracellular region of CCR5, its unsulfated analogue, and a point-mutated peptide are unable to inhibit R5-HIV-1 mediated infection, competing with the entire CCR5 in the formation of gp120-CD4-CCR5 complex. Surprisingly, these peptides show the capability of enhancing HIV-1 infection caused by X4 strains through the up regulation of both CD4 and CXCR4 receptors. PMID- 12893272 TI - Flagellin gene fliC of Xanthomonas campestris is upregulated by transcription factor Clp. AB - Clp, a homologue of cyclic AMP receptor protein (CRP), of Xanthomonas campestris regulates the expression of many genes. In this study, proteomic analysis showed the amounts of several extracellular proteins in a clp mutant to be reduced, including the flagellin encoded by fliC. Transcriptional fusion assay showed that activity of fliC promoter is reduced by 2.3-fold in clp mutant compared to the wild-type, coincident with the protein levels. The clp mutant is slightly reduced in motility; however, electron microscopy showed no significant change in the monotrichous flagellation. A fleQ homologue with conserved Clp-binding site in the upstream region is present in the fully sequenced X. campestris genome, suggesting that regulation of the flagellar genes is similar to that of Pseudomonas aeruginosa in involving Vfr, the CRP homologue, and FleQ in a cascade manner except that Vfr downregulates fleQ. Concomitant loss of flagellum and motility in fliC mutant and absence of a second homologue in the genome sequence suggest that X. campestris possesses a single flagellin gene, fliC. In addition, mutation of this gene does not affect virulence. PMID- 12893273 TI - EBNA1 may prolong G(2)/M phase and sensitize HER2/neu-overexpressing ovarian cancer cells to both topoisomerase II-targeting and paclitaxel drugs. AB - We have shown previously that the Epstein-Barr virus nuclear antigen-1 (EBNA1) can act as a transforming suppressor in the HER2/neu-overexpressing ovarian cancer cells. In the present study, by using flow cytometric analysis, we demonstrate that EBNA1 could prolong G(2)/M phase and sensitize to Taxol-induced apoptosis in the EBNA1-expressing ovarian cancer cell stable transfectants. In addition, EBNA1 could also significantly increase topoisomerase IIalpha protein expression, indicating that the up-regulation of topoisomerase IIalpha may be one of the mechanisms by which EBNA1 enhances the sensitivity of ovarian cancer cells to topoisomerase II-targeting anticancer drugs, such as VP-16 and Adriamycin. These data suggest that EBNA1 not only prolongs cell cycle at G(2)/M phase and up regulates topoisomerase IIalpha expression in HER2/neu-overexpressing ovarian cancer cells, but also increases cellular apoptosis through sensitization of cancer cells to topoisomerase II-directing anticancer drugs. PMID- 12893274 TI - Polyglutamine-expanded ataxin-1 recruits Cu/Zn-superoxide dismutase into the nucleus of HeLa cells. AB - Spinocerebellar ataxia 1 (SCA1) is an inherited neurodegenerative disorder caused by expansion of the polyglutamine stretch in ataxin-1, the SCA1 gene product. Polyglutamine expansion leads to the aggregation of ataxin-1 proteins. Superoxide dismutases (SODs) are involved in the pathogenesis of other aggregate-forming neurodegenerative diseases and are known to localize in the cytoplasm. Here, we show that Cu/Zn-SOD is translocated into the nucleus of HeLa cells in the presence of expanded ataxin-1, whereas Mn-SOD is localized in the cytoplasm: the longer the expansion of polyglutamine, the higher the level of translocation of Cu/Zn-SOD. In addition, the oxidation of intracellular proteins occurs with higher frequency in the presence of mutant ataxin-1 (82Q), suggesting that the functional activity of Cu/Zn-SOD might be decreased by mutant ataxin-1. We demonstrate that mutant ataxin-1-expressing cells encounter mitochondrial dysfunction in the conditions of oxidative stress. Our results suggest that polyglutamine-expanded ataxin-1 increases the levels of reactive oxygen species in HeLa cells. PMID- 12893275 TI - Parathyroid hormone gene with bone phenotypes in Chinese. AB - Osteoporosis is a common disorder afflicting old people. The parathyroid hormone (PTH) gene is involved in bone remodeling and calcium homeostasis, and has been considered as an important candidate gene for osteoporosis. In this study, we simultaneously tested linkage and/or association of PTH gene with bone mineral density (BMD) and bone mineral content (BMC), two important risk factors for osteoporosis. A sample of 1263 subjects from 402 Chinese nuclear families was used. The families are composed of both parents and at least one healthy daughter aged from 20 to 45 years. All the subjects were genotyped at the polymorphic BstBI site inside the intron 2 of the PTH gene (a nucleotide substitution of G to A at the position +3244). BMD and BMC were measured at the lumbar spine and the hip region via dual-energy X-ray absorptiometry (DXA). Using QTDT (quantitative trait transmission disequilibrium test), we did not find significant results for association or linkage between the PTH gene and BMD or BMC variation at the spine or hip. Our data do not support the PTH gene as a quantitative trait locus (QTL) underlying the bone phenotypic variation in the Chinese population. PMID- 12893276 TI - Differential binding properties of B7-H1 and B7-DC to programmed death-1. AB - Programmed death-1 (PD-1) is a negative regulatory receptor expressed on activated T and B cells. Two ligands for PD-1, B7-H1 (PD-L1) and B7-DC (PD-L2), have been identified, but their binding properties have not been characterized yet. In this study, we generated soluble Ig fusion proteins of these molecules and examined the kinetics and relative affinities of the interactions between B7 H1 or B7-DC and PD-1 by flow cytometry and surface plasmon resonance. The interaction of B7-DC/PD-1 exhibited a 2-6-fold higher affinity and had different association/dissociation kinetics compared with the interaction of B7-H1/PD-1. Our results suggest that the differential binding properties of B7-H1 and B7-DC may be responsible for differential contributions of these two PD-1 ligands to immune responses. PMID- 12893277 TI - Absence of superoxide dismutase activity in a soluble cellular isoform of prion protein produced by baculovirus expression system. AB - A method for expression and purification of a soluble form of histidine (HIS) tagged murine prion protein (bacMuPrP), which lacks the entire C-terminal cleavage and glycosyl phosphatidyl inositol (GPI) addition site, has been developed using a recombinant baculovirus expression system and purification with Ni-NTA agarose affinity chromatography. In mammalian sources, PrP(C) is attached to the cell membrane by a GPI anchor. However, in our system, bacMuPrP was secreted into the media, enabling its easy purification in abundance. Indirect immunofluorescence studies and immunoblot analysis localized not in cell membrane but in the perinuclear endoplasmic reticulum region in cells and is secreted into the media. Tunicamycin treatment revealed non-glycosylated proteins were secreted into the media, suggesting that glycosylation is not necessary for bacMuPrP secretion. Density-gradient sedimentation analysis demonstrated a sedimentation coefficient of secretory bacMuPrP as 2.3 S, indicating a monomeric form. Although affinity-purified PrP from mouse brain or recombinant prion protein (PrP) produced by Escherichia coli and refolded in the presence of copper has been reported to display superoxide dismutase (SOD) activity, bacMuPrP did not show SOD activity. These results suggest that bacMuPrP has a different biochemical and biophysical characterization from mammalian and bacterial-derived PrP. Furthermore, this simple expression system may provide an adequate source for structural, functional, and biochemical analyses of PrP. PMID- 12893278 TI - Functional effects of expression of wolframin-antisense transcripts in BRIN-BD11 beta-cells. AB - Wolfram syndrome is a rare condition in which the pancreatic beta-cells of patients are selectively deleted during the early years of life by a non autoimmune-mediated mechanism. The condition is associated with mutations in the gene encoding wolframin, suggesting that this protein exerts a critical, but currently unknown, function in beta-cells. We have used an antisense strategy to modulate the expression of wolframin in insulin-secreting BRIN-BD11 cells to study its function. Stably transfected clones were established expressing full length human wolframin antisense transcripts. These cells exhibited a dramatic reduction in cell proliferation rate and changes in morphology, although insulin secretion was not modified. The results imply that wolframin expression is required to sustain normal rates of beta-cell proliferation. PMID- 12893279 TI - Bimoclomol, a heat shock protein co-inducer, acts by the prolonged activation of heat shock factor-1. AB - The novel hydroxylamine derivative, bimoclomol, has been shown previously to act as a co-inducer of several heat shock proteins (Hsp-s), enhancing the amount of these proteins produced following a heat shock compared to heat shock alone. Here we show that the co-inducing effect of bimoclomol on Hsp expression is mediated via the prolonged activation of the heat shock transcription factor (HSF-1). Bimoclomol effects are abolished in cells from mice lacking HSF-1. Moreover, bimoclomol binds to HSF-1 and induces a prolonged binding of HSF-1 to the respective DNA elements. Since HSF-1 does not bind to DNA in the absence of stress, the bimoclomol-induced extension of HSF-1/DNA interaction may contribute to the chaperone co-induction of bimoclomol observed previously. These findings indicate that bimoclomol may be of value in targeting HSF-1 so as to induce up regulation of protective Hsp-s in a non-stressful manner and for therapeutic benefit. PMID- 12893280 TI - Functional expression and characterization of a sodium-dependent nucleoside transporter hCNT2 cloned from human duodenum. AB - We have cloned and functionally expressed a sodium-dependent human nucleoside transporter, hCNT2, from a CNS cancer cell line U251. Our cDNA clone of hCNT2 had the same predicted amino acid sequence as the previously cloned hCNT2 transporter. Of the several cell lines studied, the best hCNT2 transport function was obtained when transiently expressed in U251 cells. Na(+)-dependent uptake of [3H]inosine in U251 cells transiently expressing hCNT2 was 50-fold greater than that in non-transfected cells, and uptake in Na(+)-containing medium was approximately 30-fold higher than that at Na(+)-free condition. The hCNT2 displayed saturable uptake of [3H]inosine with K(m) of 12.8 microM and V(max) of 6.66 pmol/mg protein/5 min. Uptake of [3H]inosine was significantly inhibited by the purine nucleoside drugs dideoxyinosine and cladribine, but not by acyclic nucleosides including acyclovir, ganciclovir, and their prodrugs valacyclovir and valganciclovir. This indicates that the closed ribose ring is important for binding of nucleoside drugs to hCNT2. Among several pyrimidine nucleosides, hCNT2 favorably interacted with the uridine analogue floxuridine. Interestingly, we found that benzimidazole analogues, including maribavir, 5,6-dichloro-2-bromo-1 beta-D-ribofuranosylbenzimidazole (BDCRB), and 5,6-dichloro-1-beta-D ribofuranosylbenzimidazole (DRB), were strong inhibitors of inosine transport, even though they have a significantly different heterocycle structure compared to a typical purine ring. As measured by GeneChip arrays, mRNA expression of hCNT2 in human duodenum was 15-fold greater than that of hCNT1 or hENT2. Further, the rCNT2 expression in rat duodenum was 20-fold higher than rCNT1, rENT1 or rENT2. This suggests that hCNT2 (and rCNT2) may have a significant role in uptake of nucleoside drugs from the intestine and is a potential transporter target for the development of nucleoside and nucleoside-mimetic drugs. PMID- 12893281 TI - Direct superoxide anion scavenging by a disodium disuccinate astaxanthin derivative: Relative efficacy of individual stereoisomers versus the statistical mixture of stereoisomers by electron paramagnetic resonance imaging. AB - Carotenoids are a related group of greater than 600 natural compounds, irrespective of geometric- and stereoisomers, with demonstrated antioxidant efficacy. The carotenoids are broadly divided into "carotenes," or non-oxygen substituted hydrocarbon carotenoids, and "xanthophylls," oxygen-substituted carotenoids. The natural compounds are excellent singlet oxygen quenchers as well as lipid peroxidation chain-breakers; this dual antioxidant capacity is generally attributed to the activity of the polyene chain, and increases with the number of conjugated double bonds along the polyene chain length. However, the poor aqueous solubility of most carotenes and the vast majority of xanthophylls limits their use as aqueous-phase singlet oxygen quenchers and direct radical scavengers. A variety of introduction vehicles (e.g., organic solvents, cyclodextrins) have been used to introduce the insoluble carotenoids into aqueous test systems. Hawaii Biotech, Inc. (HBI) successfully synthesized a novel carotenoid derivative, the disodium disuccinate derivative of astaxanthin (3,3(')-dihydroxy beta,beta-carotene-4,4(')-dione) in all-trans (all-E) form. The novel derivative is a water-dispersible symmetric chiral molecule with two chiral centers, yielding four stereoisomeric forms: 3R,3(')R and 3S,3(')S (enantiomers), and the diastereomeric meso forms (3R,3(')S and 3(')R,3S). The individual stereoisomers were synthesized at high purity (>90% by HPLC) and compared directly for efficacy with the statistical mixture of stereoisomers obtained from the synthesis from the commercial source of astaxanthin (1:2:1 ratio of 3S,3(')S, meso, and 3R,3(')R, respectively). Direct scavenging of superoxide anion was evaluated in a standard in vitro isolated human neutrophil assay by electron paramagnetic resonance (EPR) imaging, employing the spin-trap DEPMPO. Each novel derivative was tested in pure aqueous formulation and in ethanolic formulation shown to completely disaggregate the compounds in solution. In each case, the ethanolic formulation was a more potent scavenging vehicle. No significant differences in scavenging efficiency were noted among the individual stereoisomers and the statistical mixture of stereoisomers, suggesting that the polyene chain alone was responsible for superoxide scavenging. Dose-ranging revealed that the statistical mixture of stereoisomers of the novel derivative, at millimolar (mM) concentrations, could nearly completely eliminate the superoxide anion signal generated in the activated human neutrophil assay. All ethanolic formulations of the novel derivatives exhibited increased scavenging efficiency over equimolar concentrations of non-esterified astaxanthin delivered in a dimethyl sulfoxide (DMSO) vehicle. These novel compounds will likely find utility in applications requiring aqueous delivery of a highly potent direct radical scavenger. PMID- 12893282 TI - Increase of intracellular Ca(2+) during ischemia/reperfusion injury of heart is mediated by cyclic ADP-ribose. AB - While the molecular mechanisms by which oxidants cause cytotoxicity are still poorly understood, disruption of Ca(2+) homeostasis appears to be one of the critical alterations during the oxidant-induced cytotoxic process. Here, we examined the possibility that oxidative stress may alter the metabolism of cyclic ADP-ribose (cADPR), a potent Ca(2+)-mobilizing second messenger in the heart. Isolated heart perfused by Langendorff technique was subjected to ischemia/reperfusion injury and endogenous cADPR level was determined using a specific radioimmunoassay. Following ischemia/reperfusion injury, a significant increase in intracellular cADPR level was observed. The elevation of cADPR content was closely correlated with the increase in ADP-ribosyl cyclase activity. Inclusion of oxygen free radical scavengers, 2,2,6,6-tetramethyl-1-piperidinyloxy and mannitol, in the reperfusate prevented the ischemia/reperfusion-induced increases in cADPR level and the ADP-ribosyl cyclase activity. Exposure of isolated cardiomyocytes to t-butyl hydroperoxide increased the ADP-ribosyl cyclase activity, cADPR level, and intracellular Ca(2+) concentration ([Ca(2+)](i)) and consequently resulting in cell lethal damage. The oxidant induced elevation of [Ca(2+)](i) as well as cell lethal damage was blocked by a cADPR antagonist, 8-bromo-cADPR. These results provide evidence for involvement of cADPR and its producing enzyme in alteration of Ca(2+) homeostasis during the ischemia/reperfusion injury of the heart. PMID- 12893283 TI - Identification and characterization of a new pair of immunoglobulin-like receptors LMIR1 and 2 derived from murine bone marrow-derived mast cells. AB - We have identified and characterized two mouse cDNAs in a mouse antigen stimulated bone marrow-derived mast cell cDNA library, both of which encode type I transmembrane proteins. The genes were closely mapped in the distal region of mouse chromosome 11 and expressed not only in mast cells but also widely in leukocytes. The extracellular domains of their encoded proteins contain a single variable immunoglobulin (Ig) motif sharing about 90% identity with amino acids, showing that they comprise a pair of molecules and belong to the Ig superfamily. We named these molecules leukocyte mono-Ig-like receptor1 and 2 (LMIR1 and 2). The intracellular domain of LMIR1 contains several immunoreceptor tyrosine-based inhibition motifs (ITIMs). When cross-linked, the intracellular domain was tyrosine phosphorylated and capable of recruiting tyrosine phosphatases, SHP-1 and SHP-2 and inositol polyphosphate 5-phosphatase, SHIP. LMIR2, on the other hand, contains a short cytoplasmic tail and a characteristic transmembrane domain carrying two positively charged amino acids associated with three kinds of immunoreceptor tyrosine-based activation motif (ITAM)-bearing molecules, DAP10, DAP12, and FcRgamma. These findings suggest that a new pair of ITIM/ITAM-bearing receptors, LMIR1 and 2, regulate mast cell-mediated inflammatory responses through yet to be defined ligand(s). PMID- 12893284 TI - Opioid peptides modulate the response of neurons of the superficial laminae of the rat spinal dorsal horn to GABA. AB - The modulatory effects of methionine-enkephalin (M-ENK) and selective opioid receptor agonists on GABA-activated whole-cell currents were investigated in neurons acutely dissociated from the superficial laminae of the rat spinal dorsal horn using nystatin-perforated patch recording configuration under voltage-clamp conditions. The results show that: (1). GABA acted on GABA(A) receptors and elicited inward Cl(-) currents (I(GABA)) at -60 mV; (2). M-ENK depressed I(GABA) in approximately 65% of the tested neurons and potentiated I(GABA) in approximately 15% of the neurons tested; (3). the agonists of mu-, kappa-, and delta-opioid receptors-[D-AIa(2),N-Me-Phe(4),Gly(5)-ol]-enkephalin (DAMGO), dynorphin-A (Dyn-A), and [D-Pen(2),D-Pen(5)]-enkephalin (DPDPE) also depressed the I(GABA), and the order of agonist potency was DAMGO>Dyn-A>DPDPE; and (4) naloxone blocked the inhibitory effects of M-ENK on I(GABA). The antagonists of mu-, kappa-, and delta-opioid receptors-beta-funaltrexamine (beta-FNA), nor binaltorphimine (nor-BNI), and naltrindole (NTI) prevented the DAMGO-, Dyn-A-, and DPDPE-induced depression of I(GABA). The results suggest that M-ENK downregulates I(GABA) principally through mu- and kappa-opioid receptors, and thus exerts its modulating effects indirectly on the transmission of noxious information at the spinal level. PMID- 12893285 TI - Regulation of PCNA and CAF-1 expression by the two tuberous sclerosis gene products. AB - Tuberous sclerosis is an autosomal dominant tumor suppressor gene syndrome affecting about 1 in 6000 individuals. Two genes have been shown to be responsible for this disease: TSC1, encoding hamartin and TSC, encoding tuberin. A variety of tumors characteristically occur in different organs of tuberous sclerosis patients and are believed to result from defects in cell cycle/cell size control. In this study, we performed two-dimensional gel electrophoresis with subsequent mass spectrometrical identification of protein spots after overexpression of TSC1 or TSC2. We found expression of PCNA and the p48 subunit of CAF-1 to be regulated by two tuberous sclerosis gene products. CAF-1 and PCNA interact as major regulators of chromatin assembly during DNA repair. We suggest that deregulation of the control of chromatin assembly might contribute to development of tumors in tuberous sclerosis patients and provide important new insights into the molecular development, especially since deregulation of chromatin assembly and DNA repair results in genomic instability, a hallmark of tumor development. PMID- 12893286 TI - Mutagenesis of a Gly-Ser cleavage site in MUC1 inhibits ectodomain shedding. AB - MUC1 mucin is a type 1 transmembrane glycoprotein dimer of extracellular and membrane-bound subunits. The two non-covalently associated subunits are produced from a single polypeptide chain by proteolysis at a Gly-Ser peptide bond in the endoplasmic reticulum prior to localization on the cell surface. However, once expressed on the surface, the extracellular subunit is shed from cells in the absence of the membrane-associated subunit. Previous studies implicated a cellular metalloproteinase mediating MUC1 ectodomain shedding, but no reports have delineated the site of metalloproteinase cleavage or directly assessed the role of the Gly-Ser bond in shedding. Therefore, we performed site-directed mutagenesis of the Gly-Ser site and determined the effects on MUC1 proteolysis and shedding. Ser-->Ala substitution blocked MUC1 cleavage and inhibited shedding. Equal amounts of wild type and mutant MUC1 were expressed on the cell surface, indicating that lack of shedding of the mutant molecule was not due to reduced surface localization. We conclude that the Gly-Ser peptide bond is required for MUC1 shedding. PMID- 12893287 TI - Dendrocin, a distinctive antifungal protein from bamboo shoots. AB - An antifungal protein, with a molecular weight of 20 kDa and an inhibitory action on mycelial growth in the fungi Fusarium oxysporum, Botrytis cincerea, and Mycosphaerella arachidicola, was isolated from fresh bamboo shoots. The protein, designated dendrocin, was unadsorbed on DEAE-cellulose and adsorbed on Affi-gel blue gel and CM-Sepharose. Dendrocin showed only limited similarity in N-terminal sequence to thaumatin-like proteins, unlike other thaumatin-like proteins which closely resemble each other. Its molecular weight was also lower than those of the previously reported thaumatin-like proteins. The protein was devoid of hemagglutinating and ribonuclease activities found in some antifungal proteins. PMID- 12893289 TI - The many faces of c-MYC. AB - The proto-oncogene c-MYC is implicated in various physiological processes-cell growth, proliferation, loss of differentiation, and cell death (apoptosis). Oncogenic c-MYC implies constitutive or deregulated expression of c-MYC and is associated with many human cancers often with poor prognosis. Recently, c-MYC has been implicated in the loss and dysfunction of insulin-producing beta cells in diabetes. Intriguingly, this raises the possibility that c-Myc may be a key contributor to disease, not only by deregulating cell proliferation, which is well established, but also by virtue of its opposing role in engendering apoptosis. However, given the fact that human diseases at diagnosis are generally advanced and pathologically complex, it is generally difficult to attribute a specific pathogenic role to c-MYC, or indeed any given single factor, or to assess the potential of therapies targeting individual such factors. Regulatable transgenic mouse models have shed light on these issues, have influenced our thinking about cancer, and have provided encouragement for the future development of cancer therapies based on targeting individual oncogenes such as c-MYC. Although still in its infancy, encouraging results have been reported for several approaches using gene targeting to interfere with c-MYC expression or activity both in vitro and in vivo. PMID- 12893290 TI - Rat NTE-related esterase is a membrane-associated protein, hydrolyzes phenyl valerate, and interacts with diisopropylfluorophosphate through a serine catalytic machinery. AB - The serine hydrolases constitute multi-families of proteins that include lipases, esterases, and proteases. These enzymes contain a signature motif GXSXG, in which the serine residue acts as the nucleophile and initiates catalysis. This report describes the characterization of a novel serine hydrolase from rat. This enzyme exhibits a moderate sequence identity with the neuropathy target esterase (NTE), thus is designated NTE-related esterase (NRE). Transfection with the NRE cDNA resulted in marked increases in the hydrolysis of phenyl valerate and reactivity with diisopropylfluorophosphate. Such increases, however, were markedly or completely abolished in mutants that had a substitution (Ala, Cys, Asp, or His) on the serine residue in the GXSXG motif, providing direct evidence that NRE is a serine hydrolase. By Northern blot analyses, three NRE transcripts were detected and they differed markedly in length (approximately 2.6, 4.2, and 5.0 kb). The 4.2-kb transcript was present in all organs analyzed except the testis, in which both 2.6- and 5.0-kb transcripts were detected. The testicular transcripts were completely depleted in rats treated with clofibrate, whereas the levels of NRE mRNA in the liver were markedly increased in rats treated with perfluorodecanoic acid. Both clofibrate and perfluorodecanoic acid are efficacious activators of the peroxisome proliferator activated receptor-alpha (PPAR-alpha). The differential effects on the levels of NRE mRNA suggest that these chemicals regulate the expression of NRE through mechanism(s) rather than the activation of PPAR-alpha. PMID- 12893291 TI - Novel inhibitor for prolyl aminopeptidase from Serratia marcescens and studies on the mechanism of substrate recognition of the enzyme using the inhibitor. AB - Prolyl aminopeptidase from Serratia marcescens hydrolyzed x-beta-naphthylamides (x=prolyl, alanyl, sarcosinyl, L-alpha-aminobutylyl, and norvalyl), which suggested that the enzyme has a pocket for a five-member ring. Based on the substrate specificity, novel inhibitors of Pro, Ala, and Sar having 2-tert-butyl [1,3,4]oxadiazole (TBODA) were synthesized. The K(i) value of Pro-TBODA, Ala TBODA, and Sar-TBODA was 0.5 microM, 1.6 microM, and 12mM, respectively. The crystal structure of enzyme-Pro-TBODA complex was determined. Pro-TBODA was located at the active site. Four electrostatic interactions were located between the enzyme and the amino group of Pro inhibitors (Glu204:0E1-N:Inh, Glu204:0E2 N:Inh, Glu232:0E1-N:Inh, and Gly46:O-N:Inh), and the residue of the inhibitors was inserted into the hydrophobic pocket composed of Phe139, Leu141, Leu146, Tyr149, Tyr150, and Phe236. The roles of Phe139, Tyr149, and Phe236 in the hydrophobic pocket and Glu204 and Glu232 in the electrostatic interactions were confirmed by site-directed mutagenesis, which indicated that the molecular recognition of proline is achieved through four electrostatic interactions and an insertion in the hydrophobic pocket of the enzyme. PMID- 12893292 TI - Oxidative DNA damage induced by photodegradation products of 3(')-azido-3(') deoxythymidine. AB - 3(')-Azido-3(')-deoxythymidine (AZT) is carcinogenic to experimental animals and can cause the formation of 8-oxo-7,8-dihydro-2(')-deoxyguanosine (8-oxodG) in humans and animals. To clarify the mechanism of carcinogenesis by AZT, we investigated DNA damage induced by its photodegradation products, using 32P-5(') end-labeled DNA fragments obtained from human genes. Following exposure to UVB, AZT induced DNA damage in the presence of Cu(II). Catalase inhibited DNA damage, indicating the involvement of H(2)O(2). UVB-exposed AZT plus Cu(II) induced 8 oxodG formation in a dose-dependent manner. Mass spectrum of UVB-exposed AZT demonstrated the generation of a hydroxylamine derivative. The colorimetric determination suggested that AZT was converted into the hydroxylamine derivative depending on UVB doses. UVB-exposed AZT induced double base damage at the 5(') ACG-3(') sequence, complementary to a hot spot of the p53 gene. The basic compound, hydroxylamine, showed similar site specificity. The hydroxylamine derivative produced by photodegradation and/or possible metabolism of AZT induces oxidative DNA damage, which may participate in carcinogenesis. PMID- 12893293 TI - Low resistin levels in adipose tissues and serum in high-fat fed mice and genetically obese mice: development of an ELISA system for quantification of resistin. AB - Obesity is a major risk factor for insulin resistance. Resistin, an adipocyte derived hormone-like molecule, is considered to serve as an important link between obesity and insulin resistance. However, the physiological role of resistin and the mechanism by which it neutralizes insulin action are still unclear. There are also conflicting reports that cast doubt on the cause of insulin resistance. In this study, we developed an enzyme-linked immunosorbent assay (ELISA) system for quantification of mouse resistin levels, analyzed in relation to insulin resistance. C57BL/6J mice fed high-fat diet compared with normal diet had low resistin levels (by 70%, P<0.01) in epididymal adipose tissues. Genetically obese mice, db/db and KK-A(y), had hyperinsulinemia and hyperglycemia but low resistin levels (decreases by 83 and 90%, both P<0.01) compared with C57/BL6J mice in epididymal adipose tissues. Serum resistin levels determined by Western blotting showed a similar pattern to those in adipose tissues. Resistin levels in adipose tissues correlated with serum adiponectin concentrations positively (r=0.49). Our results indicate that the novel ELISA system is suitable for measurement of resistin levels in adipose tissues. The results do not support a role for resistin in insulin resistance. PMID- 12893294 TI - BaG, a new dimeric metalloproteinase/disintegrin from the Bothrops alternatus snake venom that interacts with alpha5beta1 integrin. AB - The alpha(5)beta(1) integrin is one of the major fibronectin receptors which plays an essential role in the adhesion of normal and tumor cells to extracellular matrix. Here, we describe the isolation and characterization of a novel dimeric metalloproteinase/disintegrin, which is an inhibitor of fibronectin binding to the alpha(5)beta(1) integrin. This protein (BaG) was isolated from the venom of the South American snake Bothrops alternatus by gelatin-Sepharose affinity and anion exchange chromatography. The molecular mass of BaG was approximately 130 kDa under non-reducing conditions and 55 kDa under reducing conditions by SDS-PAGE. BaG shows proteolytic activity on casein that was inhibited by EDTA. 1,10-phenanthroline-treated BaG (BaG-I) inhibits ADP-induced platelet aggregation with an IC(50) of 190 nM. BaG-I inhibits fibronectin mediated K562 cell adhesion with an IC(50) of 3.75 microM. K562 cells bind to BaG I probably through interaction with alpha(5)beta(1) integrin, since anti alpha(5)beta(1) antibodies inhibited K562 cell adhesion to BaG-I. In addition, BaG-I induces the detachment of K562 cells that were bound to fibronectin. In summary, we have purified a novel, dimeric snake venom metalloproteinase/disintegrin that binds to the alpha(5)beta(1) integrin. PMID- 12893295 TI - Tetrahydrobiopterin is synthesized from 6-pyruvoyl-tetrahydropterin by the human aldo-keto reductase AKR1 family members. AB - Tetrahydrobiopterin (BH(4)) is a cofactor for aromatic amino acid hydroxylases and nitric oxide synthase. The biosynthesis includes two reduction steps catalyzed by sepiapterin reductase. An intermediate, 6-pyruvoyltetrahydropterin (PPH(4)) is reduced to 1(')-oxo-2(')-hydroxypropyl-tetrahydropterin (1(') OXPH(4)) or 1(')-hydroxy-2(')-oxopropyl-tetrahydropterin (2(')-OXPH(4)), which is further converted to BH(4). However, patients with sepiapterin reductase deficiency show normal urinary excretion of pterins without hyperphenylalaninemia, suggesting that other enzymes catalyze the two reduction steps. In this study, the reductase activities for the tetrahydropterin intermediates were examined using several human recombinant enzymes belonging to the aldo-keto reductase (AKR) family and short-chain dehydrogenase/reductase (SDR) family. In the reduction of PPH(4) by AKR family enzymes, 2(')-OXPH(4) was formed by 3 alpha-hydroxysteroid dehydrogenase type 2, whereas 1(')-OXPH(4) was produced by aldose reductase, aldehyde reductase, and 20 alpha-hydroxysteroid dehydrogenase, and both 1(')-OXPH(4) and 2(')-OXPH(4) were detected as the major and minor products by 3 alpha-hydroxysteroid dehydrogenases (types 1 and 3). The activities of aldose reductase and 3 alpha-hydroxysteroid dehydrogenase type 2 (106 and 35 nmol/mg/min, respectively) were higher than those of the other enzymes (0.2-4.0 nmol/mg/min). Among the SDR family enzymes, monomeric carbonyl reductase exhibited low 1(')-OXPH(4)-forming activity of 5.0 nmol/mg/min, but L xylulose reductase and peroxisomal tetrameric carbonyl reductase did not form any reduced product from PPH(4). Aldose reductase reduced 2(')-OXPH(4) to BH(4), but the other enzymes were inactive towards both 2(')-OXPH(4) and 1(')-OXPH(4). These results indicate that the tetrahydropterin intermediates are natural substrates of the human AKR family enzymes and suggest a novel alternative pathway from PPH(4) to BH(4), in which 3 alpha-hydroxysteroid dehydrogenase type 2 and aldose reductase work in concert. PMID- 12893296 TI - A 2S albumin-homologous protein from passion fruit seeds inhibits the fungal growth and acidification of the medium by Fusarium oxysporum. AB - Antimicrobial proteins have been isolated from a wide range of plant species. More recently, it has become increasingly clear that these types of proteins play an important role in the protection of plants. In this study, we investigate the presence of defense-related proteins from passion fruit (Passiflora edulis f. flavicarpa) seeds. Initially, seed flour was extracted for 2h (at 4 degrees C) with phosphate buffer, pH 5.5. The precipitate obtained between 0 and 70% relative ammonium sulfate saturation was re-dissolved in distilled water and heated at 80 degrees C for 15 min. The resulting suspension was clarified by centrifugation and the supernatant (F/0-70) was extensively dialyzed. A Sephadex G-50 size exclusion column was employed for further separation of proteins. The fraction with antifungal activity was pooled and submitted to CM-Sepharose cation exchange. Two proteins, named Pf1 and Pf2, were eluted in 0.1 and 0.2M of salt, respectively, and submitted to reverse-phase chromatography in HPLC. This fraction inhibited the growth, in an in vitro assay, of the phytopathogenic fungi Fusarium oxysporum and colletotrichum lindemuthianum and the yeast Saccharomyces cerevisiae and strongly inhibited glucose-stimulated acidification of the medium by F. oxysporum in a dose-dependent manner. The molecular masses of these proteins, referred to now as Pf1-RP and Pf2-RP, were obtained by MALDI-TOF spectrometry and corresponded to 12,088 Da for Pf1-RP and 11,930 Da for Pf2-RP. These proteins were also subjected to automated N-terminal amino acid sequencing. Sequence comparisons for the heavy subunit of Pf2-RP showed the presence of a protein with a high degree of homology to storage 2S albumins. PMID- 12893297 TI - Black tea extract supplementation decreases oxidative damage in Jurkat T cells. AB - The purpose of this study was to investigate the protective effect of black tea (BT) extract against induced oxidative damage in Jurkat T-cell line. Cells supplemented with 10 or 25 mg/L BT were subjected to oxidation with ferrous ions. Malondialdehyde (MDA) production as marker of lipid peroxidation, DNA single strand breaks as marker of DNA damage, and modification of the antioxidant enzyme activity, glutathione peroxidase (GPX) were measured. Results show the efficacy of BT polyphenols to decrease DNA oxidative damage and to affect GPX activity (P<0.05), while no effect was shown on MDA production. The succeeding investigation of the activity of caffeine and epigallocatechin gallate demonstrated their antioxidant potential with respect to the cellular markers evaluated. In conclusion, this study supports the protective effect of BT against ferrous ions induced oxidative damage to DNA and the ability of BT to affect the enzyme antioxidant system of Jurkat cells. PMID- 12893298 TI - The effect of pH on the oxidation of bovine serum albumin by hypervalent myoglobin species. AB - Bovine serum albumin (BSA) was used as a probe for the oxidation of proteins by hypervalent myoglobin species in solutions with pH from 5.3 to 7.7. The reaction between perferrylmyoglobin, *MbFe(IV)=O, and BSA was studied by activating metmyoglobin with equimolar amounts of hydrogen peroxide in the presence of BSA. A minor pH dependence was observed as judged from the formation of BSA-centered radicals, which were monitored at room temperature by electron spin resonance spectroscopy, and the formation of dityrosine. The reaction between ferrylmyoglobin, MbFe(IV)=O, and BSA was pH-dependent. BSA-centered radicals and dityrosine were formed in low levels at neutral pH and increased at low pH to the same levels as observed in the reaction of *MbFe(IV)=O with BSA. The present results demonstrate that protein-centered radicals can be formed from the non radical MbFe(IV)=O under mildly acidic conditions, and this should be taken into account when considering oxidation in cellular compartments of low pH and in meat related products. PMID- 12893299 TI - Two polycyclic aromatic hydrocarbon o-quinone reductases from a pyrene-degrading Mycobacterium. AB - Polycyclic aromatic hydrocarbon (PAH) o-quinone reductase (PQR) plays a crucial role in the detoxification of PAH o-quinones by reducing them to catechols. Two constitutive PQRs were found in cell extracts of a pyrene-degrading Mycobacterium sp. strain PYR100. The enzymes had an activity towards 9,10-phenanthrenequinone (PQ) and/or 4,5-pyrenequinone (PyQ), and the relative amounts varied with the pH of the culture media. PQR1, containing an FAD cofactor, was a monomer (20.1 kDa), and PQR2, with no flavin cofactor, was a homodimer (26.5 kDa subunits). There was no homology between the N-terminal sequences of PQR1 and PQR2. Dicumarol and quercetin inhibited PQR2 more strongly than PQR1. PQR1 had much lower specificity constants (k(cat)/K(m), 10(5)M(-1)s(-1)) for menadione (0.80) and PQ (5.19) than PQR2 (13.9 for menadione and 176 for PQ). Additionally, PQR2 exhibited a broad substrate specificity with high specificity constants for 1,4-naphthalenequinone, 1,2-naphthalenequinone, and PyQ. PMID- 12893300 TI - Catalytic oxidation of acetaminophen by tyrosinase in the presence of L-proline: a kinetic study. AB - A kinetic study of acetaminophen oxidation by tyrosinase in the presence of a physiological nucleophilic agent such as the amino acid L-proline is performed in the present paper. The o-quinone product of the catalytic activity, 4-acetamido-o benzoquinone, becomes unstable through the chemical addition of L-proline, in competition with the nucleophilic addition of hydroxide ion from water. In both cases, the catechol intermediate, 3(')-hydroxyacetaminophen, is generated, as can be demonstrated by liquid chromatography. When the effect of the presence of the nucleophilic agent on the time course of the enzymatic reaction was kinetically analyzed, it was seen to decrease the duration of the lag period and increase the steady-state rate. Rate constants for the reaction of 4-acetamido-o-benzoquinone with water and L-proline were also determined. The results obtained in this paper open a new possibility to acetaminophen toxicity, that has been attributed hitherto to its corresponding p-quinone, N-acetyl-p-benzoquinone imine. PMID- 12893301 TI - Quantitative variations of the isoforms in haptoglobin 1-2 and 2-2 individual phenotypes. AB - Haptoglobin is a hemoglobin-binding protein presenting in humans three distinct phenotypes (Hpt 1-1, Hpt 1-2, or Hpt 2-2). The Hpt 1-2 and Hpt 2-2 phenotypes are in turn represented by populations of isoforms. The relative amounts of the major isoforms of Hpt 1-2 and Hpt 2-2 were found to differ not only in different individuals, but also in the same individual before and after a physical effort. Exercise-dependent changes in the plasma concentrations of ascorbate, urate, alpha-tocopherol, retinol, and glutathione were also observed, but correlations between such changes and those of the amount for any isoform were not found. Samples of Hpt 1-2 or Hpt 2-2 were challenged with oxidants (H(2)O(2) with ferrous ions, spermine-NO, KO(2), and 3-morpholinosydnonimine), but the isoform levels were not altered. Hpt 2-2 isoforms were present in Hpt 1-2, as minor species. Furthermore, different isoforms exhibited different hemoglobin binding abilities. Thus, these parameters should also be taken into consideration in studies correlating Hpt phenotypes prevalence with pathologies or functional differences. PMID- 12893302 TI - Thiyl radicals in biosystems: inhibition of the prostaglandin metabolism by the cis-trans-isomerization of arachidonic acid double bonds. AB - This paper describes parallel and comparative experiments on the enzymatic cyclooxygenase (COX) driven conversion of arachidonic acid (AA, all-cis-5,8,11,14 eicosatetraenoic acid) into prostaglandins by using pure arachidonic acid and AA samples containing relatively small amounts of thiyl radical induced trans isomers. The experiments were performed in a liquid aqueous model system using COX-1 as well as by the in vitro feeding of VD(3)-differentiated and LPS stimulated promyelocytic HL-60 cells using the cell's own COX-2. In the model solution, all the different test methods used (oxygen consumption, ROS induced luminescence, and TMPD oxidation) indicated the greatly disproportionate, non stoichiometric inhibition of the prostaglandin metabolism by the trans-isomers. Accordingly, measurements performed in the cell system gave comparable results: both luminescence ROS detection and the ELISA test on PGE(2) expression resulted in the strong inhibition of the prostaglandin metabolism. We interpret these findings as enzyme blocking caused by just one mono-trans-isomerized double bond of AA. PMID- 12893303 TI - Protective effect of prolactin and placental lactogen on NO-induced Nb2 lymphoma cell apoptosis. AB - Nitric oxide (NO) is an important modulator involved in immune regulation. Here, we describe conditions under which NO-donors induce apoptosis on Nb2 lymphoma cells, as evidenced by decreased cell viability and increased hypodiploid DNA content determined by flow cytometry. In addition, DNA fragmentation typical of apoptosis was shown by agarose gel electrophoresis. This apoptosis was accompanied by a significant increase of caspase-3-like enzymatic activity. Both ovine prolactin (oPRL) and ovine placental lactogen (oPL) exerted a protective effect on the NO-donor-induced apoptosis. Furthermore, dexamethasone (Dex) induced cell death was also associated with caspase-3-like activity and oPL had the same potency as oPRL in its protective effect on Dex-induced apoptosis of Nb2 cells. PMID- 12893304 TI - Quantitative model of small molecules uptake after in vitro cell electropermeabilization. AB - Electropermeabilization of the cell membrane is a phenomenon caused by exposure of the cell to electric pulses. Permeabilization depends on pulse duration, pulse amplitude, the number of pulses delivered, and also on other experimental conditions. With these parameters properly chosen, the process of permeabilization is reversible and cells return to their normal physiological state. This article describes the development of a model of diffusion-driven transmembrane transport of small molecules caused by electropermeabilization. The process of permeabilization is divided into a short permeabilizing phase that takes place during the pulse, and a longer resealing phase that begins after the end of the pulse. Because both phases of permeabilization are important for uptake of molecules into cells, most of the effort is focused on the optimization of parameters that influence the flow between intracellular and extracellular space. The model describes well the transmembrane transport caused by electropermeabilization, allowing to study the uptake of molecules as a function of elapsed time, voltage and pulse duration. In addition, our results show that the shapes of the curves of cell permeabilization and survival as functions of pulse amplitude can to a large extent be explained by cell size distribution. PMID- 12893305 TI - Electrochemical monitoring of the interaction of doxorubicin with nicotinamide and Fe(III) ions under aerobic and anaerobic conditions. AB - The interaction of doxorubicin with Fe(III) ions and nicotinamide (NA) has been followed by square-wave voltammetry, cyclic voltammetry and UV-VIS. spectroscopy techniques at aerobic and anaerobic conditions. Fe(III)-doxorubicin complex gives a 1-electron reversible step at -0.494 V and a shoulder at 580 nm. Further, the Fe(III)-doxorubicin complex was found to be more stable at aerobic conditions. In the presence of NA, an intermediate (NA-Fe(III)-DQ) forms at -0.462 V under aerobic conditions. Because of the formation of this intermediate, nicotinamide may reduce the cardiotoxic effect of doxorubicin and cause to its detoxification. PMID- 12893306 TI - Effective gating charge of ion channels induced by toxin syringomycin E in lipid bilayers. AB - To elucidate the voltage gating of syringomycin E (SRE) ion channels in lipid bilayers, the effective gating charge q was measured under different conditions. It was shown that q and its sign are dependent on membrane surface charge, dipole potential, and the outer potential (Delta phi). The q values were positive for charged bilayers and negative for uncharged bilayers bathed in the same 0.1 M NaCl solutions. Effects of dipole modifying agents on the gating properties of SRE channels were measured. In uncharged bilayers, addition of phloretin resulted in an increase of q values. For charged bilayers, the presence of RH-421 or 6 ketocholestanol leads to the reverse in the sign of q from positive to negative. The q values were potential-dependent at higher negative voltages with charged membranes bathed in solutions with high salt concentrations. It is concluded that lipid molecules participating in the SRE channel structure contribute to channel formation work due to Coulomb and dipolar interactions with the electric field applied to a membrane. The potential dependence of q is explained by interactions of charged and uncharged lipids with SRE molecules in the channels. PMID- 12893307 TI - Electrochemical study of heavy metals and metallothionein in yeast Yarrowia lipolytica. AB - The bioaccumulation of heavy metals (cadmium, nickel, cobalt and zinc) and the effect of these metals on the production of metallothionein and metallothionein like proteins (MT) in Yarrowia lipolytica was studied by electrochemical methods. The concentrations of heavy metals were determined by differential pulse voltammetry (DPV). A combination of the constant current chronopotentiometric stripping analysis (CPSA) and adsorptive transfer stripping technique (AdTS) was used to determine the content of MT in cells. Both the bioaccumulation of heavy metals and the production of MT in different cell compartments of Y. lipolytica exposed to heavy metals were monitored. The LD(50) of each metal was determined from the number of viable cells in yeast cultures: LD(50)Cd (37.5 microM), LD(50)Ni (570 microM), LD(50)Co (700 microM), and LD(50)Zn (1800 microM). The highest concentrations of heavy metals were found in the cell wall and membrane debris while the lowest concentrations were detected in the cytoplasm. Cadmium and nickel showed the most significant effect on the production of MT. This study provides new insights into the ecophysiology of microorganisms and demonstrates the potential use of these electrochemical methods in biotechnology. PMID- 12893308 TI - Electrochemical study on the keto-enol tautomerization of p-hydroxyphenylpyruvic acid in aqueous solution. AB - The keto-enol tautomerization of p-hydroxyphenylpyruvic acid (pHPP) in aqueous solutions and the complexation reaction between enolic pHPP and boric acid have been studied by electrochemical techniques including linear sweep voltammetry (LSV), pulse voltammetry, and cyclic voltammetry (CV), combining with UV spectrometry. Electrochemical techniques reveal that in aqueous solution, there are two tautomers of pHPP: enolic form and ketonic form; the former exists mainly in freshly prepared pHPP solution, and the latter exists mainly in equilibrium solution. Both enolic and ketonic pHPP are electroactive. The electrochemical oxidation of enolic pHPP gives rise to two anodic waves, I(a) and II(a), while the electrochemical oxidation of ketonic pHPP only results in the observation of the second wave II(a). The oxidation process I(a) is revealed to be associated with the quasi-reversible, two-electron two-proton oxidation of "C=C"group at the side chain of enolic pHPP, and the oxidation process II(a) is proposed to result from the irreversible oxidation of phenolic hydroxyl group. It is observed that in aqueous solution, enolic pHPP can quickly complex with boric acid to yield enol-borate complex that can also oxidize at a glassy carbon electrode to yield an anodic wave. PMID- 12893309 TI - A short-term NaCl exposure increases the Na(+) conductance of outward-rectified cation currents in the pith cells of sweet pepper. AB - The regulatory role of pith cells in the stem in Na(+) recirculation in sweet pepper was investigated by evaluating the transport characteristics of the plasma membrane of this cell type and comparison with those of root cells. Ion conductivity and Na(+) permeability of the plasma membranes of protoplasts of both cell types were studied with the patch-clamp technique in the whole-cell configuration, before and after addition of NaCl to the bath medium. Protoplasts of both pith and root cells showed outward rectifying currents with a reversal potential (V(r)) near to the equilibrium potential of K(+) (EK). Addition of NaCl to the bath medium caused a stronger shift of the reversal potential, V(r), in pith protoplasts than in root protoplasts, indicating that the outward rectified currents are permeable to Na(+), especially in the pith cells. After plant exposure to exogenous NaCl via the nutrient solution for 1 week, V(r) in the root cells was closer to EK than in the control plants and hardly shifted upon addition of Na(+). This indicated that the net permeability of the OR channel complement in the plasma membrane to Na(+) was lower following exposure to Na(+). V(r) in the pith protoplasts, on the other hand, shifted significantly more than in the control plants, suggesting an increase of the permeability to Na(+). Moreover, the Na(+) channel blocker amiloride blocked the currents in this cell type. It is concluded that pith cells have appropriate features of outward rectified currents to enable Na(+) accumulation or release when NaCl is present in or removed from the nutrient medium. Probably, exogenous NaCl even induced expression and formation of Na(+)-permeable channels in pith cells. PMID- 12893310 TI - Voltammetric and spectroelectrochemical characterization of a water-soluble viologen polymer and its application to electron-transfer mediator for enzyme free regeneration of NADH. AB - A water-soluble polyxylylviologen (PXV(2+)) was characterized with a view to making use of it as a redox electron-transfer (ET) mediator. Cyclic voltammetric and spectropotentiometric studies showed (i) that PXV(2+) gives two redox waves centering at -0.40 and -0.83 V (vs. Ag/AgCl (3.3 mol dm(-3) KCl)) and (ii) that the lifetime of its monocation radical (PXV(+.)) is two orders of magnitude greater than that of the well-utilized dimethyl viologen monocation radical. Subsequently, the reaction of the PXV(2+/+.) couple with NAD(+) was evaluated in the similar manners as above. On the basis of this evaluation and the bioluminescence assay using bacterial NADH/FMN oxidoreductase and luciferase, it was shown (i) that the PXV(2+/+.) couple functions as a useful electron-transfer mediator and (ii) that PXV(+.) reacts with NAD(+), leading to generation of the enzymatically active NADH, in the absence of any reductases. PMID- 12893311 TI - Evaluation of redox mediators for amperometric biosensors: Ru-complex modified carbon-paste/enzyme electrodes. AB - The properties of reagentless amperometric biosensors are mainly governed by the interaction of the used redox enzyme and the redox mediators used to facilitate the electron-transfer reaction. Both the used redox mediators and the redox enzymes differ concerning their hydrophilicity and their properties within the matrix of a carbon-paste electrode. Since there is no general procedure which is applicable for any enzyme in combination with any redox mediator, optimisation is necessary for each possible combination. Three approaches for the development of biosensors were investigated using carbon-paste electrodes enriched with redox mediator as a base in all sensor architectures. A class of redox mediators with the common formula Ru(LL)(2)(X)(2) (where LL are 1,10-phenantroline or 2,2' bipyridine type ligands, and X is an acido ligand) was investigated. In the first approach, enzymes were integrated into the carbon paste; in the second, the enzymes were adsorbed on the surface of the mediator-containing carbon-paste electrode and held in place by a Nafion film; and in the third approach, enzymes were entrapped in polymer films, which were electrochemically deposited onto the electrode's surface. The properties of the obtained biosensors strongly depend on the sensor architecture and the specific features of the used enzyme. Thus, our investigation using three different sensor architectures can provide valuable information about the possible interaction between a specific enzyme and a redox mediators with specific properties. PMID- 12893312 TI - Effects of ELF and static magnetic fields on calcium oscillations in islets of Langerhans. AB - Several experimental studies have produced contradictory results on the effects of extremely low frequency (ELF) magnetic fields on cellular processes involving calcium ions. Furthermore, the few positive results have not been independently replicated. In most of these studies, isolated cells were used. Our study used mouse islets of Langerhans, in which very regular oscillations of calcium concentration can be observed at length. These oscillations are sustained by processes that imply energetic and inter-intracellular communication. Various magnetic fields were applied, either sinusoidal at different frequencies (50 Hz or multiples of the natural oscillation frequency) at 0.1 or 1 mT or static at 1 mT. Islets were also exposed to "cyclotron resonance" conditions. There was neither alteration of the fundamental oscillation frequency nor the degree of organisation under all exposure conditions. Using this sensitive model, we could not show new evidence of alterations of calcium processes under exposure to various magnetic fields. PMID- 12893314 TI - Photoelectric effects on chlorophyll fluorescence of photosystem II in vivo. Kinetics in the absence and presence of valinomycin. AB - Fluorescence induction curves (F(t)) in low intensity 1s light pulses have been measured in leaf discs in the presence and absence of valinomycin (VMC). Addition of VMC causes: (i) no effect on the initial fluorescence level Fo and the initial (O-J) phase of F(t) in the 0.01-1 ms time range. (ii) An approximately 10% decrease in the maximal fluorescence Fm in the light reached at the P level in the O-J-I-P induction curve. (iii) Nearly twofold increase in the rate and extent of the F(t) rise in the J-I phase in the 1-50 ms time range. (iv) A 60-70% decrease in the rise (I-P phase) in the 50-1000 ms time range with no appreciable effect, if at all, on the rate. System analysis of F(t) in terms of rate constants of electron transfer at donor and acceptor sides have been done using the Three State Trapping Model (TSTM). This reveals that VMC causes: (i) no, or very little effect on rate constants of e-transfer reactions powered by PSII. (ii) A manifold lower rate constant of radical pair recombination (k(-1)) in the light as compared to that in the control. The low rate constant of radical pair recombination in the reaction center (RC) in the presence of VMC is reflected by a substantial increase in the nonzero trapping efficiency in RCs in which the primary quinone acceptor (Q(A)) is reduced (semi-open centers). This causes an increase in their rate of closure and in the overall trapping efficiency. Data suggest evidence that membrane chaotropic agents like VMC abolish the stimulation of the rate constant of radical pair recombination by light. This light stimulation that becomes apparent as an increase in Fo has been documented before [Biophys. J. 79 (2000) 26]. It has been ascribed to effects of (changes in) local electric fields in the vicinity of the RC. The decrease of the I-P phase is attributed to a decrease in the photoelectric trans-thylakoid potential in the presence of VMC. Such effects have been hypothesized and illustrated. PMID- 12893313 TI - Methotrexate cytotoxicity on MCF-7 breast cancer cells is not altered by exposure to 25 Hz, 1.5 mT magnetic field and iron (III) chloride hexahydrate. AB - The action of electromagnetic fields (EMF) on different pathways related to cell physiology, proliferation, toxicity of chemicals, gene expression, etc., are currently being investigated although the results are still not conclusive and even conflicting. In laboratory and animal studies, EMF has been found to produce a great variety of effects such as: increase in ornithine decarboxylase activity in breast, increase in beta-galactosidase gene expression and oncogene transcription after exposure to 50/60 Hz. Animal studies have shown that the use of EMF can enhance drug delivery across biological barriers (rat abdominal skin), using benzoic acid as the drug candidate. It has been reported by different authors that pulsed EMF (PEMF) can produce alterations in antineoplastic drugs potency. In the present study, we investigated the effects of PEMF on methotrexate cytotoxicity in MCF-7 breast cancer cells and the effects with simultaneous exposure to FeCl3. The data presented in the current report indicate that PEMF (25 Hz, 1.5 mT) do not induce modulation of the action of methotrexate (with and without iron-III) in MCF-7 cells when they are exposed to PEMF for 2 h/day during 3 days. PMID- 12893315 TI - Modeling of ionic relaxation around a biomembrane disk. AB - A simplified Brownian dynamics model and the corresponding software implementation have been developed for the simulation of electrolyte dynamics on the mesoscopic scale. In addition to direct control simulations, the model system has been verified by a quantitative comparison with the Debye-Huckel theory. As a first application, the model was used to simulate ionic relaxation processes following abrupt intramembrane charge rearrangements in the case of a disk shaped membrane. In addition to its general implications, the obtained properties of the relaxation kinetics confirm the assumptions of the theory of the so-called suspension method, a technique capable of tracing molecular charge motions of membrane proteins in three dimensions. PMID- 12893316 TI - Electrical energy required to form large conducting pores. AB - This study computes the contribution of the externally induced transmembrane potential to the energy of large, highly conductive pores. This work was undertaken because the pore energy formulas existing in the literature predict qualitatively different behavior of large pores: the original formula proposed by Abidor et al. in 1979 implies that the electrical force expanding the pore increases linearly with pore radius, while later extensions of this formula imply that this force decreases to zero for large pores. Starting from the Maxwell stress tensors, our study derives the formula for the mechanical work required to deform a dielectric body in an ionic solution with steady-state electric current. This formula is related to a boundary value problem (BVP) governing electric potentials and fields in a proximity of a pore. Computer simulations yield estimates of the electrical energy for pores of two different shapes: cylindrical and toroidal. In both cases, the energy increases linearly for pore radii above approximately 20 nm, implying that the electrical force expanding the pore asymptotes to a constant value for large pores. This result is different from either of the two energy formulas mentioned above. Our study traces the source of this disagreement to approximations made by previous studies, which are suitable only for small pores. Therefore, this study provides a better understanding of the energy of large pores, which is needed for designing pulsing protocols for DNA delivery. PMID- 12893317 TI - Imaging of enzyme activity by scanning electrochemical microscope equipped with a feedback control for substrate-probe distance. AB - The enzymatic activity of diaphorase (Dp) immobilized on a solid substrate was characterized using a scanning electrochemical microscope (SECM) with shear force feedback to control the substrate-probe distance. The shear force between the substrate and the probe was monitored with a tuning fork-type quartz crystal and used as the feedback control to set the microelectrode probe close to the substrate surface. The sensitivity and the contrast of the SECM image were improved in the constant distance mode (distance, 50 nm) with the shear force feedback compared to the image in the constant height mode without the feedback. By using this system, the SECM and topographic images of the immobilized diaphorase were simultaneously measured. The microelectrode tip used in this study was ground aslant like a syringe needle in order to obtain the shaper topographic images. This shape was also effective for avoiding the interference during the diffusion of the enzyme substrates. PMID- 12893319 TI - Dose rate in external beam radiotherapy for prostate cancer: an overlooked confounding variable? PMID- 12893320 TI - Effective testosterone suppression for patients with prostate cancer: is there a best castration? PMID- 12893321 TI - Characteristics of bacterial colonization and urinary tract infection after indwelling of double-J ureteral stent. AB - OBJECTIVES: To investigate the natural history of bacterial colonization on the stent and in the urine after different periods of indwelling ureteral stent placement. METHODS: A total of 57 double-J stents from 52 patients (21 men and 31 women, mean age 52 years, range 20 to 79) were examined. In all patients, short term (2 to 3 days) antimicrobial therapy was administered at the time of stenting. Regular urinalysis and urine culture were performed until the stent was removed. The stents were removed by aseptic manipulation, and the proximal and distal tip segments of the stents were obtained. Three culture specimens were acquired from each stent segment (inner surface washing suspension, outer surface washing suspension, and from the stent tip itself). RESULTS: Bacterial colonies were found in 44% (25 of 57) of the stents. Of the multiple pathogens identified, Enterococcus species (6 of 25) was the most common, followed by Escherichia coli (5 of 25). After short-term antibiotic prophylaxis, the bacteria did not colonize within the first 2 weeks of stent placement. However, the colonization rate increased as the duration of the stent placement lengthened. Colonization of the stent was followed by colonization of the urine. CONCLUSIONS: The results of our study demonstrated that bacterial colonization at the stent began 2 weeks after indwelling stent placement and that stent colonization preceded urine colonization. The rate of colonization increased with longer periods of stenting. PMID- 12893322 TI - Evaluation of overall costs of currently available small flexible ureteroscopes. AB - OBJECTIVES: To perform a meta-analysis of the currently available data regarding the durability of flexible ureteroscopes to establish cost estimates for the purchase and use of five currently available, smaller than 9F, ureteroscopes. Healthcare costs have become increasingly germane to the determination of disease management strategies. Improved ureteroscope technology has expanded the role of these instruments. However, the initial purchase costs and high maintenance costs have become problematic with these fragile instruments. METHODS: Ureteroscope durability data on the Storz 11274AA, Olympus URF-P3, Wolf 7325.172, ACMI AUR-7, and ACMI DUR-8 were collected from three prior studies. Combining the durability data and cost data regarding the initial purchase price and maintenance costs of these instruments, we calculated the overall costs associated with the use of each of the ureteroscopes for 25, 50, 75, and 100 cases during the first year (warranties included) and with subsequent use. RESULTS: The variability in the costs associated with the use of the currently available smaller than 9F ureteroscopes was significant. The initial instrument purchase price, durability, repair costs, and associated warranties all contributed to large discrepancies in the cost of performing ureteroscopy. In this model, during the first year of ownership, the projected cost of performing 100 ureteroscopic cases varied by a difference of 95% depending on the ureteroscope used. CONCLUSIONS: Physicians and institutions that perform ureteroscopy should strongly consider the purchase price, durability, repair cost, and associated warranties before the purchase of small flexible ureteroscopes. PMID- 12893323 TI - Laparoscopic upper pole partial nephrectomy with concomitant en bloc adrenalectomy. AB - OBJECTIVES: To report our experience with laparoscopic partial nephrectomy for renal tumor with concomitant adrenalectomy. An upper pole renal tumor may contiguously involve the adrenal gland, requiring concomitant adrenalectomy. Although commonly performed in the setting of laparoscopic radical nephrectomy, concomitant adrenalectomy has not been described during laparoscopic partial nephrectomy. METHODS: Four patients with an upper pole renal tumor and suspected adrenal involvement underwent laparoscopic partial nephrectomy with concomitant ipsilateral adrenalectomy. Preoperative three-dimensional computed tomography revealed the renal tumor to be closely abutting the adrenal gland in 3 patients and a 4-cm adrenal mass in 1 patient. The mean renal tumor size was 3.2 cm (range 1.4 to 6.6). To maintain oncologic principles, our transperitoneal laparoscopic technique excises the adrenal gland en bloc with the renal tumor. As such, adrenalectomy is performed first, followed by partial nephrectomy, incorporating hilar control, tumor excision, and sutured renal reconstruction. RESULTS: All four procedures were performed without open conversion or intraoperative complications. The mean renal warm ischemia time was 36 minutes, estimated blood loss 169 mL, total operating time 3.9 hours, and hospital stay 3.2 days. One patient developed a transient urinary leak postoperatively. Pathologic examination of the renal tumor revealed renal cell carcinoma (n = 1), dystrophic calcification with ectopic bone formation (n = 1), adult mesoblastic nephroma (n = 1), and subcapsular heterotopic adrenal cortex with cyst (n = 1), all with negative surgical margins. Pathologic examination of the adrenal gland revealed adenoma in 1 case and a normal adrenal gland without malignant involvement in 3 cases. All patients were disease free at last follow-up (mean 6.2 months, range 2 to 12). CONCLUSIONS: In patients with an upper pole renal tumor and radiologically suspected adrenal involvement, laparoscopic partial nephrectomy with concomitant adrenalectomy can be performed efficaciously respecting oncologic principles. PMID- 12893324 TI - Renal artery pseudoaneurysm after partial nephrectomy: three case reports and a literature review. AB - OBJECTIVES: To report 3 cases of renal artery pseudoaneurysm (RAP) after partial nephrectomy and review the literature to discuss their unique presentation, clinical course, and management. RAPs are a rare clinical entity. They are a well documented source of hemorrhage in association with renal biopsy, renal trauma, renal transplant, and percutaneous procedures; however, little is known about their presentation, pathophysiology, and management after nephron-sparing surgery. METHODS: Between 2000 and 2002, 698 partial nephrectomies were performed at our institution for treatment of suspected localized renal malignancy. A retrospective review of these cases revealed that 3 patients developed a postoperative RAP. These cases were reviewed along with the current medical literature available from the MEDLINE database to characterize this clinical entity further. RESULTS: Three patients developed RAP after 698 uncomplicated partial nephrectomies were performed (0.43% incidence). Illustrating a classic presentation, the patient in case 1 was successfully treated with selective embolization after presenting with symptoms 3 weeks postoperatively. The patient in case 2 was treated with elective partial selective embolization for an asymptomatic lesion that later resolved spontaneously. In case 3, the patient presented early during the hospital course and required urgent selective embolization. CONCLUSIONS: RAP after nephron-sparing surgery is a rare, but potentially life-threatening, condition that is often difficult to diagnose and requires a high index of suspicion. Treatment should be tailored to each patient's clinical presentation to minimize morbidity and maximize renal conservation. All urologists should be aware of this potential postoperative complication and prepare accordingly. PMID- 12893325 TI - Three-dimensional ultrasonography: an objective outcome tool to assess collagen distribution in women with stress urinary incontinence. AB - OBJECTIVES: To examine the distribution of periurethral collagen by three dimensional ultrasonography (3D US) and to incorporate this technology into a practical treatment decision tree for women with stress urinary incontinence desiring collagen injection. METHODS: Forty-six women who received periurethral collagen injection were assessed with 3D US to document the position and volume of collagen around the urethra. Patients with a good clinical response were observed with serial 3D US scans. Women with no persistent improvement who showed a low volume or asymmetric distribution of collagen were offered repeat injections. When the patient had no improvement despite symmetric or circumferential distribution of collagen with good volume retention, another anti incontinence treatment was recommended. Group 1 was composed of women who had a good clinical outcome and group 2 of those who did not. A statistical comparison between the two outcome groups was done to determine whether any particular distribution pattern was associated with an increased likelihood of clinical success. RESULTS: Of the 46 patients, 21 (46%) were satisfied with their continence after their last periurethral collagen injection, with a median follow up of 14.0 months (range 2 months to 4.25 years). A significantly greater proportion of patients in group 1 had circumferentially distributed collagen on 3D US compared with those in group 2 (62% versus 20%, P = 0.006). CONCLUSIONS: Circumferential distribution of collagen around the urethra is associated with a higher likelihood of clinical success. The assessment of periurethral collagen by 3D US provides an affordable, noninvasive, objective outcome measure that may aid in treatment planning. PMID- 12893326 TI - Comparative efficacy and safety of transdermal oxybutynin and oral tolterodine versus placebo in previously treated patients with urge and mixed urinary incontinence. AB - OBJECTIVES: To compare the efficacy and safety of an oxybutynin transdermal delivery system (OXY-TDS) and oral, long-acting tolterodine (TOL-LA) with placebo in previously treated patients with urge or mixed urinary incontinence. METHODS: After withdrawal of their current antimuscarinic therapy, 361 adult patients were randomized to 12 weeks of double-blind, double-dummy treatment with twice weekly OXY-TDS 3.9 mg/day, daily TOL-LA 4 mg, or placebo. Evaluations included change from baseline in patient urinary diary symptoms, incontinence-specific quality of life, and safety. RESULTS: OXY-TDS 3.9 mg/day and TOL-LA 4 mg/day significantly reduced the number of daily incontinence episodes (median change -3 OXY-TDS and 3 TOL-LA versus -2 placebo; P <0.05), increased the average void volume (median change 24 and 29 mL versus 5.5 mL, P <0.01), and improved quality of life (incontinence impact questionnaire [IIQ] total score, P <0.05; Urogenital Distress Inventory Irritative Symptom subscale, P <0.05) compared with placebo. The most common adverse event for OXY-TDS was localized application site pruritus (14% versus 4% placebo) accompanied by a low incidence of systemic side effects (eg, dry mouth 4.1%). Anticholinergic adverse events occurred with greatest frequency during TOL-LA treatment (dry mouth 7.3% versus 1.7% placebo, P <0.05). CONCLUSIONS: OXY-TDS and TOL-LA are effective and comparable treatments for patients with urge and mixed incontinence. OXY-TDS improves systemic safety with regard to anticholinergic side effects. Local skin irritation occurs in some OXY TDS patients. PMID- 12893327 TI - Usefulness of tissue polypeptide antigen in the follow-up of bladder cancer. AB - OBJECTIVES: To evaluate the sensitivity and specificity of serum tissue polypeptide antigen (TPA) as a tumor marker for monitoring tumor recurrence in a prospective Phase II study. METHODS: This study consisted of 421 subjects allocated to three groups: 241 patients treated for bladder cancer between 1991 and 1995, 126 subjects with a previous history of bladder tumor but with no recurrence at the time of study enrollment, and 54 healthy volunteers. Soluble TPA concentrations were measured by radioimmunoassay. RESULTS: On the basis of receiver operating characteristic plot analysis, the normal test threshold was set at 63 U/L. For bladder cancer detection, the TPA specificity was 73% and sensitivity was 45%. TPA levels correlated with tumor stage (P <0.01), histologic grade, nodal stage, and metastatic stage and with urinary cytology in univariate analysis. In the subsets of patients with a history of bladder cancer and normal TPA levels, 34.9% experienced a relapse compared with 37.5% of patients with elevated TPA levels (no statistically significant difference). Among the patients with normal TPA levels who experienced recurrence, the mean disease-free interval was 30.6 months compared with 25.1 months in patients with elevated TPA levels (no statistically significant difference). Multivariate analysis showed that TPA was not an independent prognostic biomarker for tumor relapse. CONCLUSIONS: In patients with bladder cancer, the TPA serum level correlated with initial tumor stage and grade, but it was not useful for detecting or monitoring relapses. PMID- 12893328 TI - Tumor-associated trypsin inhibitor as a prognostic factor during follow-up of bladder cancer. AB - OBJECTIVES: To evaluate the prognostic value of tumor-associated trypsin inhibitor (TATI) in the serum and urine of patients in follow-up for urinary bladder cancer. METHODS: Serum and urine samples were taken during follow-up of 157 patients with transitional cell carcinoma of the bladder who were monitored by cystoscopy and cytology in 1986 to 1987. Initially, 117 (75%) of the 157 tumors were superficial. At the time of sampling, 93 patients (59%) had no detectable tumor and 48 (31%) had a superficial, and 16 (10%) an invasive, tumor. Cancer-specific survival was evaluated in 1998. RESULTS: During follow-up, 35 patients (22%) died of bladder cancer. An elevated TATI concentration in the serum (21 microg/L or more) was associated with a significantly shorter survival (P <0.001) compared with a normal value. Multivariate analysis showed that serum TATI and detectable cancer at sampling were independent prognostic factors (P <0.001 and P = 0.002, respectively), and age, grade, urine cytology findings, and urine TATI were not. CONCLUSIONS: Serum TATI is an independent prognostic factor in transitional cell carcinoma and is potentially useful for the identification of patients with an adverse prognosis. PMID- 12893329 TI - A new technique for creation of a sigmoid neobladder for urinary reconstruction: clinical outcome in 42 men. AB - OBJECTIVES: To present the results of a new technique for construction of a sigmoid neobladder after radical cystectomy. METHODS: Forty-two men, 43 to 74 years of age (mean age 62 +/- 8), underwent radical cystectomy for invasive bladder cancer and had a sigmoid neobladder created using a modification of the standard ureterointestinal anastomosis with rotation of the reservoir. The mean follow-up of this retrospective study was 35 +/- 20 months (range 6 to 90). The blood chemistry was analyzed routinely during follow-up, and complications, continence, and voiding pattern were assessed at regular intervals. RESULTS: There were 21 early complications in 14 patients: persistent urinary leakage in 2; wound infection in 9; prolonged ileus in 4; leakage from the colonic anastomosis in 2; and pulmonary embolus in 2. One patient each had venous thrombus of the lower extremities and methicillin-resistant Staphylococcus aureus colitis. One patient died in the perioperative period of massive pulmonary embolus, yielding a mortality rate of 2.4%. Eleven late complications occurred in 9 patients. Stenosis of the ureterointestinal anastomosis developed in 2 of 82 anastomoses, stenosis of urethral anastomosis in 4, and neobladder-ureteral reflux in 1. A reservoir stone was found in 1 patient, and adenoma and adenocarcinoma of the reservoir occurred in 1 patient each. One patient developed symptomatic hyperchloremic metabolic acidosis, and routine electrolyte evaluation revealed a slight metabolic acidosis in 6 patients. Hypovitaminosis B(12) did not occur in any patients. Of the 42 patients, 40 could void volitionally without catheterization. Patient satisfaction on continence, by day and night, was 77.3% and 72.7%, respectively. The mean neobladder capacity and the mean residual urine volume was 375 +/- 116 mL (range 234 to 624) and 68 +/- 86 mL (range 4 to 365), respectively. CONCLUSIONS: The modified sigmoid neobladder provides satisfactory results after radical cystectomy. PMID- 12893330 TI - Relief by botulinum toxin of voiding dysfunction due to benign prostatic hyperplasia: results of a randomized, placebo-controlled study. AB - OBJECTIVES: To evaluate the therapeutic role of botulinum toxin injection in men with benign prostatic hyperplasia. METHODS: Men with benign prostatic hyperplasia were enrolled in a randomized, placebo-controlled study. After a baseline evaluation, each participant received 4 mL of solution injected into the prostate gland. Patients in the control group received saline solution and patients in the treated group received 200 U of botulinum toxin A. The outcome of each group was evaluated by comparing the symptom scores, serum prostate-specific antigen concentration, prostate volume, postvoid residual urine volume, and peak urinary flow rates. RESULTS: Thirty consecutive patients were enrolled. No local complications or systemic side effects were observed in any patient. After 2 months, 13 patients in the treated group and 3 in the control group had subjective symptomatic relief (P = 0.0007). In patients who received botulinum toxin, the symptom score was reduced by 65% compared with baseline values and the serum prostate-specific antigen concentration by 51% from baseline. In patients who received saline, the symptom score and serum prostate-specific antigen concentration were not significantly changed compared with the baseline values and 1-month values. Follow-up averaged 19.6 +/- 3.8 months. CONCLUSIONS: Botulinum toxin injected into the prostate seems to be a promising approach for the treatment of benign prostatic hyperplasia. It is safe, effective, and well tolerated. Furthermore, it is not related to the patient's willingness to complete treatment. PMID- 12893333 TI - Lower urinary tract symptoms of men seeking medical care--comparison of symptoms found in the clinical setting and in a community study. AB - OBJECTIVES: To investigate which lower urinary tract symptoms were most influential in causing men to seek medical care. METHODS: We evaluated the International Prostate Symptom Score (IPSS) and quality-of-life (QOL) score of 235 outpatients having lower urinary tract symptoms and 242 participants in a community-based study of Japanese men aged 50 to 79 years old. RESULTS: Although the proportion of outpatients in the severe IPSS category (IPSS 20 to 35) was greater than that in the participants of the community-based study in each age decade, the proportion in the moderate IPSS category (IPSS 8 to 19) in both groups overlapped each other. On the other hand, the distribution of QOL scores was considerably different, with only a small portion of overlap in each age decade. Although scores for both voiding symptoms (incomplete emptying, intermittency, weak stream, and hesitancy) and storage symptoms (increased frequency, urgency, and nocturia) were significantly greater in outpatients than in study participants in each age decade, the difference was more obvious for voiding symptoms than for storage symptoms. CONCLUSIONS: The QOL score appeared to show more pronounced differences between men in a clinic setting and those in a community setting than the IPSS category. Voiding symptoms may affect medical care-seeking behavior through QOL impairment in Japanese men. PMID- 12893334 TI - Internet use by patients with prostate cancer undergoing radiotherapy. AB - OBJECTIVES: To analyze the amount of Internet use, type of Internet use, and factors that predict Internet use by patients with prostate cancer. METHODS: A questionnaire evaluating the use of the Internet was given to 295 patients undergoing radiotherapy for prostate cancer in three different settings: an academic center (AC), a Veterans Affairs center (VA), and a community hospital. The study included 171 AC patients (58%), 104 VA patients (35%), and 20 community hospital patients (7%). RESULTS: The response rate to the questionnaire was greater than 99%. Overall, 128 (43%) of 295 patients had a home computer. This differed among the institutions, with 64% of AC patients and only 5% of VA patients having a home computer (P <0.0001). Overall, 94 (32%) of 295 patients used the Internet to access cancer information, with 48% of AC patients and 8% of VA patients using the Internet (P <0.0001). Seventeen percent of black patients (n = 113) had a home computer compared with 62% of white patients (n = 175; P <0.0001). Equal access to computers did not influence Internet use because, among those patients who had access to computers, black patients used the Internet less than did white patients (32% versus 61%, P <0.0001). Men 65 years of age or younger were more likely to use the Internet (45%) than those older than 65 years (28%; P = 0.004). Patients used many sites to access different types of information. CONCLUSIONS: A significant number of patients with prostate cancer access the Internet to obtain cancer information. Urologists and radiation oncologists should be familiar with this important resource to help patients access appropriate material. PMID- 12893335 TI - Evolution of free, complexed, and total serum prostate-specific antigen and their ratios during 1 year of follow-up of men with febrile urinary tract infection. AB - OBJECTIVES: To determine the serum values of free, complexed, and total prostate specific antigen (PSA) and their ratios in men with febrile urinary tract infection (UTI) during 1 year of follow-up. METHODS: A total of 54 men were prospectively enrolled in the study, and serum samples were obtained at the acute stage of the UTI and after 1, 3, 6, and 12 months. RESULTS: Most men had a rise (median 3.1, 7.2, and 14.1 ng/mL, respectively) in free PSA (fPSA), complexed PSA (cPSA), and total PSA (tPSA) during the acute phase of the UTI. After 1 month, fPSA rapidly decreased to levels that were maintained during the rest of the follow-up period, and cPSA and tPSA declined more slowly. At 1, 3, and 6 months after the infection the fPSA/tPSA and fPSA/cPSA ratios were abnormal in one half to two thirds of the men (median ratio 11%, 15%, and 16% and 17%, 24%, and 24%, respectively), comparable to findings in patients with prostate cancer. CONCLUSIONS: These results confirmed that the prostate is involved in men with febrile UTI. The low fPSA/tPSA and fPSA/cPSA ratios in combination with sustained elevations of cPSA and tPSA for up to 6 months after UTI could be falsely interpreted as a sign of prostate cancer. The prolonged elevation of cPSA indicates a long-lasting inflammation of the nonadenomatous parts of the prostate. PMID- 12893336 TI - Detection of alpha-methylacyl-coenzyme A racemase in postradiation prostatic adenocarcinoma. AB - OBJECTIVES: To assess the utility of alpha-methylacyl-coenzyme A racemase (AMACR), also known as P504S, immunohistochemistry in the detection of postradiation prostatic adenocarcinoma in surgical specimens. Pathologic diagnosis of postradiation prostate cancer is difficult because of the radiation induced cytologic changes in benign and malignant epithelial cells. AMACR/P504S is a recently identified molecular marker for prostatic adenocarcinoma. It has been demonstrated that AMACR is overexpressed in the vast majority of prostatic adenocarcinoma cases by cDNA microarray, RNA analysis, Western blotting, and immunohistochemistry. METHODS: A total of 80 prostate glands, including 40 irradiated prostate specimens (28 with adenocarcinoma and 12 benign prostates) and 40 nonirradiated prostate specimens (20 with adenocarcinoma and 20 benign prostates), were examined. The specimens were obtained after salvage radical prostatectomy (n = 25), transurethral resection (n = 4), or needle biopsy (n = 11). All samples were immunohistochemically analyzed for AMACR. RESULTS: All 48 carcinoma cases (28 of 28 irradiated and 20 of 20 nonirradiated specimens) showed strongly positive AMACR/P504S immunostaining. AMACR immunostaining was negative for all irradiated (n = 12) and nonirradiated (n = 20) benign prostates, as well as the irradiated benign glands adjacent to carcinoma. 34betaE12 confirmed the presence of basal cells in all benign prostates (32 of 32) and the absence of basal cells in carcinoma (0 of 48). CONCLUSIONS: Our results demonstrate that AMACR is a highly specific and sensitive indicator of postradiation prostate cancer. AMACR immunostaining facilitates the challenging differentiation between prostatic adenocarcinoma and radiation-induced atypia in benign prostatic epithelium and may be of exceptional value in limited needle biopsies. PMID- 12893337 TI - Tamsulosin reduces the incidence of acute urinary retention following early removal of the urinary catheter after radical retropubic prostatectomy. AB - OBJECTIVES: To determine the efficacy of tamsulosin in preventing acute urinary retention following early catheter removal after radical retropubic prostatectomy. METHODS: Between February 2000 and October 2000, cystography was performed on postoperative day 7 after radical retropubic prostatectomy by a single surgeon (group 1). Between September 2001 and August 2002, cystography was performed on postoperative day 8 after radical retropubic prostatectomy by the same surgeon (group 2). The protocol for performing cystography and assessment of extravasation was similar for both groups. Tamsulosin 0.4 mg was administered 3 days before and 4 days after cystography for all men in group 2. RESULTS: Of 179 cystograms in group 1, 135 (75%) revealed no extravasation, and the catheters were removed in 130 of these cases. Of 246 cystograms in group 2, 230 (93.5%) revealed no extravasation, and the catheters were removed in 229 of these cases. A significantly greater proportion of men in group 2 had no extravasation (P = 0.0007). The incidence of acute urinary retention in groups 1 and 2 was 10% and 2.6%, respectively (P = 0.0018). The incidence of anastomotic stricture was not significantly different between the two groups. CONCLUSIONS: Our data strongly suggest that tamsulosin significantly reduces the risk of acute urinary retention after attempts at early catheter removal following radical retropubic prostatectomy. Therefore, we recommend administering a 7-day course of tamsulosin therapy when attempting to remove the urinary catheter before postoperative day 8. PMID- 12893338 TI - Radical retropubic versus laparoscopic prostatectomy: a prospective comparison of functional outcome. AB - OBJECTIVES: To evaluate and compare prospectively the functional results (urinary continence and erectile function) after retropubic and laparoscopic radical prostatectomy. METHODS: From 1998 to 2001, 300 patients underwent radical prostatectomy, 70 by the retropubic and 230 by the laparoscopic approach. All of them completed a confidential, self-administered questionnaire regarding urinary continence and erectile function before and 1, 3, 6, and 12 months after surgery. The results were analyzed separately for day and nighttime urinary continence and by neurovascular bundle status and age for erectile function. RESULTS: No differences were found between the two groups except for the duration of bladder catheterization (7.8 days for the retropubic and 5.8 for the laparoscopic approach, P = 0.0006). Before surgery, 296 patients (98.7%) were continent and 290 (96.6%) were potent. One year after surgery, diurnal urinary continence increased progressively to 66.7% and 71.6% (P >0.05), nocturnal continence to 66.7% and 87.1% (P = 0.008), and potency rates to 30% and 41% (P >0.05) for the retropubic and the laparoscopic approaches, respectively. After preservation of one and two neurovascular bundles, the potency rate increased to 27% and 46% and 44% and 53% for the retropubic and laparoscopic approaches, respectively (P >0.05). For patients younger than 60 years old with bilateral neurovascular bundle preservation, the potency rates were 72% and 81% (P >0.05). CONCLUSIONS: The two approaches showed similar outcomes regarding continence and erectile function 1 year after surgery. However, the recovery of nocturnal continence seemed to occur earlier after the laparoscopic approach. It is necessary to conduct additional studies to determine the potential benefits of the laparoscopic approach regarding nocturnal continence. PMID- 12893339 TI - Positive prostate biopsy laterality and implications for staging. AB - OBJECTIVES: To examine the effect of including positive prostate biopsy information in palpation staging (2002 system) and the influence of this information on freedom from biochemical failure (bNED). Prostate biopsy laterality status (unilateral versus bilateral positive) is part of clinical staging using American Joint Commission on Cancer criteria, but is rarely used. METHODS: From April 1, 1989 to September 30, 1999, 1038 patients with palpable T1 T3Nx-0M0 prostate cancer were treated with three-dimensional conformal radiotherapy alone. Kaplan-Meier bNED curves were compared using the log-rank test. The Cox proportional hazards regression model of bNED was used for multivariate analysis. RESULTS: The median follow-up was 46 months. The proportion of patients with bilateral positive biopsies by palpation category T1c was 24%, by T2a was 17%, by T2b was 26%, by T2c was 65%, and by T3 was 53%. No statistically significant difference was noted in bNED on the basis of biopsy laterality status for the palpation T stages T1c, T2a, T2b, or T3. A statistically significant difference in the 5-year bNED in the T2c stage was found; those with unilateral positive biopsies fared worse (46% versus 74%, respectively, P = 0.04). CONCLUSIONS: Inclusion of positive biopsy laterality status into clinical staging causes stage migration without reflecting a change in outcome and should not be used. PMID- 12893340 TI - Seminal vesicle involvement after radical prostatectomy: predicting risk factors for progression. AB - OBJECTIVES: To evaluate and study the factors for progression after radical prostatectomy with seminal vesicle invasion in a cohort of European and American patients. METHODS: The data of 137 patients with isolated seminal vesicle invasion who underwent radical retropubic prostatectomy between 1988 and 2001 were analyzed. The Gleason score of the specimen, presence of capsular invasion, and surgical margin status were noted. Follow-up data were obtained through routine serum prostate-specific antigen (PSA) assays and digital rectal examinations. A defining criterion for progression was a postoperative serum PSA level greater than 0.2 ng/mL or any postoperative radiotherapy or hormonal treatment. Kaplan-Meier analysis was used to determine the actuarial biochemical recurrence-free likelihood, and the log-rank test was used to compare these results. Differences were considered statistically significant when the P value was less than 0.05. RESULTS: After a mean follow-up of 4.9 years (range 0.9 to 13.4), 70 patients (51.9%) had progression. The biochemical 5-year progression free survival rate was 33.8%. In univariate and multivariate analyses, only preoperative PSA level (P = 0.001) and Gleason score of the specimen (P = 0.01) were independent predictors of progression. Neither capsular invasion nor positive surgical margins predicted progression. When an analysis was performed according to the major Gleason grade of the radical prostatectomy specimen, Gleason grade 5 was associated with a worse prognosis compared with Gleason grade 3 and 4 (P = 0.01). The mean time to progression was 20.5, 17.1, and 10.1 months for Gleason grade 3, 4, and 5, respectively. CONCLUSIONS: Seminal vesicle invasion after radical prostatectomy has historically been associated with a poor prognosis. However, in the present study, seminal vesicle invasion was associated with a 34% rate of freedom from progression at 5 years after surgery alone. Preoperative PSA and Gleason score of the radical prostatectomy specimen were independent factors for progression in the present study, which described the largest patient group to date. The Gleason grade of the radical prostatectomy specimen distinguished among different times to progression. PMID- 12893341 TI - Operative management of rectal injuries during laparoscopic radical prostatectomy. AB - OBJECTIVES: To present our experience in the management of rectal injuries during laparoscopic radical prostatectomy. METHODS: All patients underwent bowel preparation and received perioperative antibiotics. If rectal injury was suspected, digital rectal examination was performed, aided by bubbling air into the rectum if required. When the diagnosis was confirmed, the hole was closed in two layers of absorbable sutures. A fat flap was developed from the omentum (in the transperitoneal approach) or the perirectal fat (in the extraperitoneal approach) and placed on the suture line. Anal dilation was performed. After surgery, broad-spectrum antibiotics and a low-residue diet were prescribed. RESULTS: Of 300 patients who underwent surgery between May 1998 and June 2002, 6 (2%) had a rectal injury. The first patient had received neoadjuvant hormonal therapy. Five cases were in the transperitoneal approach and were closed using omental fat and 1 was in the extraperitoneal approach with a perirectal fat flap. The first patient presented with a rectourethral fistula and was treated with catheterization for 1 month and a diverting colostomy. The others were detected intraoperatively and were treated laparoscopically. In the second patient, a diverting colostomy was performed at the end of surgery using a separate incision. Patients resumed oral intake within 2 to 7 days and were discharged from the hospital between 6 and 18 days postoperatively. No wound infection was noted. CONCLUSIONS: Rectal injuries during laparoscopic radical prostatectomy can be identified and managed intraoperatively without requiring conversion. Double layered closure reinforced by a fat flap resulted in an uneventful recovery. PMID- 12893342 TI - Laparoscopic radical prostatectomy--an analysis of factors affecting operating time. AB - OBJECTIVES: Although laparoscopic radical prostatectomy (LRP) is accomplished within 2 to 3 hours by experienced surgeons, less is known about the operating times (OTs) for recently trained surgeons or the influence of additional factors. As of November 2001 at our institution, two senior surgeons had each performed more than 100 cases of LRP and two junior surgeons had each performed fewer than 30. We prospectively studied the next 100 consecutive LRPs to assess the factors influencing the OT. METHODS: Transperitoneal LRPs were performed by two senior (n = 62) and two junior surgeons (n = 38) with random case assignment. We assessed body mass index, prostate size, prior abdominal surgery, androgen deprivation, surgeon experience, procedures in addition to LRP, lymph node dissection, nerve sparing, and sural nerve grafting as potential predictors of the OT. RESULTS: Prostate weight, androgen deprivation, and prior abdominal surgery did not significantly affect the OT, but grade 1 obesity increased the OT by an average of 38 minutes. The mean OT by surgeon experience was 214 minutes for seniors and 347 minutes for juniors (P <0.001). By procedure type, the OT ranged from 180 minutes for LRP only by seniors to 459 minutes for LRP plus lymph node dissection plus sural nerve grafting by juniors. Lymph node dissection and sural nerve grafting significantly increased the OT by 46 and 101 minutes, respectively, and nerve sparing did not. For each combination of procedures, seniors averaged significantly shorter times than did juniors. A multiple regression model with stepwise selection showed that prostate weight, sural nerve grafting, pelvic lymph node dissection, use of a surgical robot, and surgeon experience significantly affected the OT. CONCLUSIONS: The results of this prospective study of 100 cases of LRP showed that the OT for senior surgeons averaged 2 to 3 hours, but less experienced surgeons, and additional procedures, add significantly to the OT. PMID- 12893343 TI - An eight-month clinical study of LA-2575 30.0 mg: a new 4-month, subcutaneous delivery system for leuprolide acetate in the treatment of prostate cancer. AB - OBJECTIVES: To investigate the safety, efficacy, and pharmacokinetics of a new 4 month subcutaneous depot of leuprolide acetate in patients with prostate cancer. METHODS: Ninety patients diagnosed with adenocarcinoma of the prostate were enrolled in an open-label, multicenter study. LA-2575 30.0 mg was administered subcutaneously once every 4 months for 8 months. The primary efficacy parameter was a serum testosterone level of 50 ng/dL or less. The pharmacokinetics of leuprolide acetate were analyzed in the first 24 enrolled patients. The values are reported as the mean +/- standard error. RESULTS: Of 90 enrolled patients, 82 (91%) completed the 8-month study. Eight patients voluntarily withdrew from the study for the following reasons: nonmedical reasons (n = 3), treatment-related adverse events (n = 3), disease progression (n = 1), and cardiovascular disease (n = 1). By day 28, 85 (94%) of the 90 patients had achieved a serum testosterone level less than 50 ng/dL. At study completion, 88 (98%) of the 90 patients had a testosterone value less than the castrate level (mean 12.4 +/- 0.8 ng/dL), with 81 (90%) at less than 20 ng/dL. From baseline to month 6, the mean luteinizing hormone level had decreased from 7.51 +/- 0.69 mIU/mL to 0.12 +/- 0.02 mIU/mL. The mean prostate-specific antigen level had decreased 90% from 13.2 +/- 2.0 ng/mL at baseline to 1.3 +/- 0.3 ng/mL at 8 months. No clinically significant flare reactions were observed. The most common treatment-related adverse event was mild hot flashes. CONCLUSIONS: LA-2575 30.0-mg depot consistently produced and maintained safe and effective suppression of serum testosterone, with total serum testosterone concentrations well below the medical castrate level of less than 50 ng/dL. PMID- 12893344 TI - Laparoscopic retroperitoneal lymph node dissection for clinical stage I nonseminomatous germ cell testicular cancer: a long-term update. AB - OBJECTIVES: To assess retrospectively the long-term cancer control in patients undergoing laparoscopic retroperitoneal lymph node dissection (RPLND) in the management of clinical Stage I nonseminomatous germ cell testicular tumors. METHODS: A retrospective review of 29 patients undergoing laparoscopic RPLND was performed. All patients had clinical Stage I nonseminomatous germ cell testicular tumor, with vascular invasion and/or embryonal carcinoma in the orchiectomy specimen. A modified template dissection was performed. Patients with retroperitoneal metastases were offered two cycles of chemotherapy. A modified follow-up schedule was used in patients with pathologically negative nodes. RESULTS: Lymph nodes were negative in 17 of 29 patients. Of these 17 patients, 15 had no recurrence and were free of disease with 5.8 years of follow-up. Two patients had recurrence, one in the chest, and one biochemically, and both were free of disease after chemotherapy. Twelve of 29 patients had lymph nodes with metastatic testicular cancer. Ten of these patients underwent adjuvant chemotherapy and were free of disease with 6.3 years of follow-up. One patient had a biochemical recurrence after positive RPLND and was salvaged with chemotherapy. One patient was observed after positive RPLND and was free of disease with 4.9 years of follow-up. The only long-term complication was retrograde ejaculation in 1 patient. CONCLUSIONS: Laparoscopic RPLND is a safe, minimally invasive treatment option in patients with clinical Stage I nonseminomatous germ cell testicular tumor. The cancer control appears to be similar, with minimal morbidity compared with the open procedure. PMID- 12893345 TI - Testicular and epididymal percutaneous sperm aspiration in men with either obstructive or nonobstructive azoospermia. AB - OBJECTIVES: To investigate our experience with epididymal and testicular sperm obtained by percutaneous aspiration to determine the effectiveness of these procedures in patients with obstructive or nonobstructive azoospermia. We also report on the pooled outcome of intracytoplasmic sperm injection, performed at six in vitro fertilization centers, using sperm from these two patient populations. METHODS: A method of percutaneous aspiration of sperm was developed to facilitate diagnostic biopsy and therapeutic aspiration. A total of 265 aspirations were performed for diagnostic and therapeutic purposes from August 1995 through July 2001. RESULTS: Mature sperm were found in 34 (47%) of 73 men with nonobstructive azoospermia, who underwent testicular sperm aspiration for diagnostic purposes. In 56 procedures for men with nonobstructive azoospermia who underwent testicular sperm aspiration for intracytoplasmic sperm injection, an adequate number of viable sperm was obtained from 46 (82%), with an oocyte fertilization and clinical pregnancy rate of 55% and 44%, respectively. In men with obstructive azoospermia who underwent percutaneous epididymal sperm aspiration, viable sperm were found in 91 (97%) of 94 procedures, with an oocyte fertilization and clinical pregnancy rate of 58% and 39%, respectively. Sperm were obtained in 18 (100%) of 18 procedures in men with obstructive azoospermia who underwent testicular sperm aspiration, with a corresponding fertilization and clinical pregnancy rate of 51% and 38%, respectively. A small (less than 1 cm in diameter) subcapsular hematoma was the only complication. CONCLUSIONS: Percutaneous aspiration of sperm is an effective, safe, and reproducible means of obtaining adequate sperm for diagnosis or intracytoplasmic sperm injection in patients with obstructive or nonobstructive azoospermia. Our experience indicates that similar results can be obtained in both patient groups. PMID- 12893346 TI - Percutaneous cystolithotomy using a laparoscopic entrapment sac. AB - OBJECTIVES: To describe a percutaneous approach that uses laparoscopic techniques and technology to achieve intact removal of bladder stones after augmentation cystoplasty. METHODS: Percutaneous stone removal using a laparoscopic entrapment sac was performed in 4 patients with augmented bladders. Under endoscopic guidance, a 10-mm laparoscopic trocar was placed percutaneously into the augmented bladder using the previous suprapubic tube site. The stones were then maneuvered into a laparoscopic entrapment sac and extracted intact without lithotripsy. RESULTS: Percutaneous removal of the entire stone burden (up to seven stones in 1 patient) was achieved in 3 of 4 patients. The total operative time was less than 1 hour in each of these cases. Partial conversion to open cystolithotomy was required in 1 patient, because of tearing of the entrapment sac. Three of 4 cases were performed on an outpatient basis and less than 24 hours of catheter drainage was required in all but 1 patient. CONCLUSIONS: Percutaneous cystolithotomy using a laparoscopic entrapment sac is a safe, useful, and minimally invasive modification of contemporary percutaneous techniques. In patients with augmented bladders, application of this technique may minimize the risk of residual fragments and obviate the need for adjuvant lithotripsy. PMID- 12893348 TI - Fetal bony pelvis in the bladder exstrophy complex: normal potential for growth? AB - OBJECTIVES: To provide the first look at the bony histologic features of fetuses with the exstrophy complex, specifically evaluating the endochondral ossification, stage of development, and microscopic potential for normal growth. METHODS: Three fetuses between 28 and 30 weeks of gestation, one with classic bladder exstrophy, one with cloacal exstrophy, and one control, were obtained from France. The bony pelves were dissected and preserved in formalin, and multiple representative sections were sliced from all pelvic areas: pubis, ischium, ilium, and sacrum. These slices were sequentially processed as slides, stained with hematoxylin-eosin, and evaluated microscopically for histologic changes, developmental stage, and degree of endochondral ossification. RESULTS: All slides from the three specimens showed cartilage analogue with endochondral ossification. Histologically the exstrophy specimens were identical to the control and appeared completely normal; bone development was occurring at an expected rate with the potential for continued normal growth. CONCLUSIONS: These new findings illustrate that fetal bone in the exstrophy complex displays normal microscopic growth patterns and unhindered endochondral ossification at 28 weeks of gestation, well beyond the embryologic period. With no evident microscopic bony defect, the gross bony anomalies in exstrophy should be surgically correctable, leading us to conclude that early reapproximation of the physiologic shape of the pelvis could lead to more normal gross bone growth, decreased shortage of bone, and a more appropriate distribution of the mechanical and developmental forces on a closed, normally functioning pelvic ring. PMID- 12893349 TI - Applications of FloSeal in nephron-sparing surgery. AB - Hemostasis can be a challenging problem to control laparoscopically. We advocate the use of FloSeal Matrix. FloSeal can be used in a variety of open and laparoscopic procedures, including laparoscopic or open partial nephrectomy. FloSeal was highly effective in stopping bleeding in a bloody surgical field. PMID- 12893350 TI - A novel technique to prevent inadvertent sheath migration. AB - Inadvertent trocar-sheath migration is annoying to the surgeon and may result in serious visceral injury. Herein is described a novel technique to prevent such migration. A 0.25-in. Penrose drain is wrapped around the trocar-sheath unit as a collar to prevent sheath migration. PMID- 12893351 TI - Modified "tubeless" percutaneous nephrolithotomy using a tail-stent. AB - INTRODUCTION: "Tubeless" percutaneous renal surgery has previously been described at our institution. Avoiding external nephrostomy tube drainage significantly decreases the pain and morbidity of percutaneous renal surgery. However, internal drainage with a double-J ureteral stent still results in bothersome bladder symptoms, especially in younger male patients. In addition, removal of the stent requires uncomfortable office cystoscopy. We report a technical enhancement of our "tubeless" technique that uses a novel tail-stent for internal drainage. TECHNICAL CONSIDERATIONS: A tubeless approach to any renal surgery should only be attempted in select uncomplicated cases. The exclusion criteria should include operative times longer than 2 hours, three or more percutaneous accesses, significant perforations or disruptions of the collecting system, significant residual stone burden, and significant bleeding. In these instances, and when second-look nephroscopy is desired, traditional external nephrostomy tube drainage should be used. In our select cases, in lieu of the standard double-J stent, we place a 7F/3F tail-stent with the string attached exiting the urethral meatus. Care is taken in correct placement to avoid having the tail of the stent exiting the meatus. CONCLUSIONS: In our initial 4 patients, tail-stents were successfully placed. The pain scores were low and stent symptoms appeared minimal. All stents were easily removed without the use of cystoscopy. Fluoroscopic visualization of the 3F tail is poor, and positioning of the tail can be difficult. Nevertheless, this modification appears feasible and safe with excellent patient satisfaction. PMID- 12893352 TI - Persistent mullerian duct syndrome with crossed testicular ectopia. PMID- 12893353 TI - Management of metastatic malignant melanoma of the bladder. AB - Recombinant interleukin-2 (IL-2) has demonstrated antitumor activity and durable clinical responses in patients with metastatic melanoma. Careful screening and selection of appropriate patients has improved the safety profile of IL-2 administration. Gross hematuria would ordinarily preclude the safe delivery of IL 2. We report a case of metastatic melanoma to the bladder presenting with hematuria. A complete resection was performed and subsequently allowed the administration of high-dose, bolus IL-2. The combination of resection and IL-2 therapy resulted in a partial response maintained for more than 18 months. Symptomatic bladder melanoma should be aggressively treated to allow for systemic immunotherapy, which can provide durable responses. PMID- 12893354 TI - Juvenile granulosa cell tumor of the testis. AB - Juvenile granulosa cell tumor of the testis is a rare type of intermediate stromal cell tumor. Only 11 cases of juvenile granulosa cell tumor of the testis have been reported to the national tumor registry as of December 2001. We report an additional case of primary juvenile granulosa cell tumor of the testis. The benign lesion was managed successfully with orchiectomy, and follow-up computed tomography revealed no evidence of recurrence at 6 months. The histopathologic features, treatment, and prognosis of this unusual tumor are reviewed. PMID- 12893355 TI - Nephrectomy after radiofrequency ablation-induced ureteropelvic junction obstruction: potential complication and long-term assessment of ablation adequacy. AB - Little information is available concerning the morbidity of radiofrequency ablation (RFA) or the evolution of an RFA lesion over time. We report our findings in a kidney removed 1 year after RFA of a 2.3-cm renal tumor. After RFA, the patient experienced flank pain, followed by hydronephrosis, ureteropelvic junction obstruction, and eventual loss of function in the treated kidney. Nephrectomy revealed no residual renal cell carcinoma. RFA can completely destroy renal cell carcinoma in situ without histologic evidence of persistence or recurrence for up to 1 year after treatment. Care must be taken to avoid concurrent damage to the collecting system. PMID- 12893357 TI - Acquired bladder diverticula can mimic obstructive extrabiliary jaundice. AB - A rare case that relates benign prostatic hyperplasia-associated bladder diverticula and obstructive uropathy to extrabiliary obstructive jaundice in an older patient is presented. Immediate decompression of the bladder allowed for prompt restoration of the biliary drainage and normalization of the creatinine within a few days. A hepatobiliary etiology was discarded through prompt radiologic and serologic testing along with computed tomography-guided liver biopsy. Long-term management included open suprapubic prostatectomy and diverticulectomy. This unusual case expands the amount of sound anatomic and pathophysiologic links between urinary and extraurinary manifestations. PMID- 12893356 TI - Multiple intravesical linear staples identified during surveillance cystoscopy after laparoscopic nephroureterectomy. AB - Laparoscopic nephroureterectomy has faced technical criticism secondary to the unknown effects of urine-exposed intravesical or intraureteral staples. Although not reported, staples exposed to urine theoretically create a nidus for encrustation and stone formation. We present a case of a 66-year-old man after left laparoscopic ureterectomy for ureteral malignancy. Surveillance cystoscopy at 6 months revealed a nearly complete intravesical titanium staple line without encrustation. Successful transurethral removal of all staples with standard cystoscopic graspers revealed no immediate, or delayed, sequelae. PMID- 12893358 TI - Luteinizing hormone-releasing hormone agonists and meningioma: a treatment dilemma. AB - The relative contraindication of hormonal therapy for patients with prostate cancer and a history of meningioma has not been widely emphasized. Using immunohistochemistry to determine the presence of hormone receptors in meningioma specimens proved potentially valuable in 2 patients with biochemical recurrence after prostatectomy who were being considered for androgen deprivation therapy. These cases also highlight the need for caution against assuming that skull-based intracranial growths in patients receiving hormonal therapy for prostate cancer are metastatic lesions rather than hormonally induced primary tumors. PMID- 12893359 TI - Endoscopic basket extraction of a urethral foreign body. AB - The presence of a foreign body in the genitourinary tract represents a urologic challenge that often requires prompt intervention. We describe an endoscopic approach that was used for removal of a foreign body located in the prostatic urethra of a schizophrenic man. A nitinol stone basket was used as an effective means for extraction of a large metal screw from the urethra of this patient. PMID- 12893360 TI - Ureteral stenosis as the sole manifestation of Wegener's granulomatosis. AB - In Wegener's granulomatosis, necrotizing lesions are typically located in the upper and lower respiratory tract and kidneys, and ureteral involvement is uncommon. We report 2 cases in which intrinsic ureteral stenosis was the sole manifestation of this small-vessel vasculitis. Excisional surgery evidenced characteristic granulomatous inflammation that allowed adjuvant elective medical treatment. Urologists, nephrologists, and internists should be aware of this atypical presentation of Wegener's granulomatosis. Thorough clinical and biologic assessments are warranted in the initial workup of isolated intrinsic ureteral stenosis. PMID- 12893361 TI - Rare case of metastatic prostate adenocarcinoma to the pituitary. AB - We report a rare case of metastatic prostate adenocarcinoma to the pituitary gland. The patient had lung and bone metastases when he presented with bitemporal hemianopsia. He was also receiving total androgen blockade at that time. Magnetic resonance imaging showed an intrasellar mass, and biopsy confirmed metastatic prostate cancer. Radiotherapy, in the form of intensity-modulated radiotherapy, was delivered to the intrasellar mass. The patient responded well to radiotherapy. The mechanisms of spread to the pituitary gland and treatment options, including intensity-modulated radiotherapy, are discussed. PMID- 12893362 TI - Isolated testicular polyarteritis nodosa mimicking testicular neoplasm-long-term follow-up. AB - Polyarteritis nodosa is a systemic vasculitis characterized by segmental necrotizing lesions of medium and small-size arteries. Rarely, the inflammatory process is isolated and involves a single organ without systemic manifestations. We describe a patient with isolated polyarteritis nodosa of the testis who presented with a testicular mass mimicking primary testicular tumor. The postoperative laboratory evaluation was negative. Long-term follow-up, without systemic treatment, showed no evidence of recurrence in the remainder of the testis or development of systemic vasculitis. PMID- 12893363 TI - Carbon tetrachloride-induced nephrotoxicity and protective effect of betaine in Sprague-Dawley rats. AB - OBJECTIVES: To observe the changes in the antioxidative defense enzymes and to detect the alterations of renal microscopy after carbon tetrachloride (CCl4) administration in rats and to investigate the possible protective effects of betaine against CCl4-induced renal damage. METHODS: Thirty-two adult Sprague Dawley rats were divided into four groups as follows: control group, betaine group, CCl4 group, and CCl4 + betaine group. CCl4 was given subcutaneously at 1 mL/kg. In the CCl4 + betaine group, rats were pretreated with betaine, then exposed to CCl4 at the same dose. Betaine group rats received concentrated betaine solution. The rats were killed and the kidneys taken for enzyme analyses and histologic examination. Glutathione peroxidase, superoxide dismutase, and catalase activities were measured in right kidney homogenates. Left kidneys were processed for light microscopic evaluation. RESULTS: In the CCl4-treated group, significant increases in kidney superoxide dismutase and catalase activities and significant decrease in glutathione peroxidase activity were observed (P <0.01). These changes were found to be normalized in the CCl4 + betaine group. Betaine did not change the enzyme activities. Exposure to CCl4 resulted in glomerular and tubular alterations in the renal cortex. These alterations were found to be prevented by betaine pretreatment. CONCLUSIONS: These results indicate that exposure to CCl4 leads to renal damage in rats and betaine exerts an improvement on nephrotoxic effects of CCl4. PMID- 12893364 TI - Muscarinic receptor subtypes mediate vasorelaxation in isolated horse deep dorsal penile vein. AB - OBJECTIVES: To investigate the effect of acetylcholine (ACh) on horse deep dorsal penile vein and to characterize the muscarinic receptor subtypes involved in this response. METHODS: Vein rings were mounted in an organ bath chamber, and the isometric tension was recorded. RESULTS: In phenylephrine-contracted veins, ACh (1 nM to 1 microM) induced endothelium-dependent relaxation. The muscarinic receptor antagonist, atropine, produced parallel rightward shifts of the ACh response curves (pA2 = 10.04; pK(B) = 9.98). Carbachol (10 nM to 100 microM) also evoked relaxation in the vein segments, but showed a lower potency and similar relaxation to that induced by ACh. Pirenzepine, the high, intermediate, and low affinity antagonist for M1, M3, and M2 receptors, respectively, inhibited ACh and carbachol-induced relaxation, yielding pA2 values of 7.51 and 7.37, and pK(B) values of 7.38 and 7.28, respectively. Methoctramine, a high-affinity M2 antagonist, showed no significant effect on the response to ACh. However, a high affinity M3 antagonist, pFHHSiD, potently blocked the relaxation induced by carbachol and ACh, yielding pA2 and pK(B) values of 7.72 and 7.70 for pFHHSiD against ACh, respectively, and of 7.77 and 7.65 against carbachol, respectively. CONCLUSIONS: These results indicate that ACh induces an endothelium-dependent relaxation in horse deep dorsal penile vein. The antagonist profile suggests that M3 muscarinic receptors mediate ACh-induced relaxation in this tissue. PMID- 12893365 TI - Detection of human telomerase reverse transcriptase mRNA in urine of patients with bladder cancer: evaluation of an emerging tumor marker. AB - OBJECTIVES: To assess the value of the telomerase enzyme as a bladder cancer detection marker, we investigated the expression of the catalytic subunit of the complex (human telomerase reverse transcriptase [hTERT]) in the urine of patients with malignant or benign urinary lesions, as well as of healthy individuals, and compared the results with urine cytology. METHODS: Spontaneously voided samples were obtained from two groups of subjects: group 1, 146 previously untreated patients with a histologic diagnosis of transitional cell carcinoma or other urothelial neoplasm; and group 2, 128 control individuals, either healthy or with a nonmalignant bladder disease. Total RNA extracts from sedimented urothelial cells were analyzed by a reverse transcriptase-polymerase chain reaction assay for the presence of a 146-bp hTERT transcript. Urine samples were also examined by standard cytology. RESULTS: Expression of hTERT was detected in 134 (92%) of 146 patients with bladder cancer, and only 64 (44%) yielded a positive result by cytology (P <0.001). The sensitivity advantage of the former technique became particularly evident in the detection of low-grade transitional cell carcinoma (93% versus 28%, P <0.001). Accordingly, the negative predictive value of the molecular assay was markedly greater than the one calculated for cytologic screening (91% versus 60%). On the other hand, both methods were at least 96% specific, with their positive predictive indexes exceeding 94%. CONCLUSIONS: Our findings suggest that the assessment of hTERT expression in urine sediments represents a reliable tool for the detection of primary urothelial neoplasms, equally specific, yet far more sensitive, than conventional cytology. PMID- 12893366 TI - Prognostic significance of CA 125, CD44, and epithelial membrane antigen in renal cell carcinoma. AB - OBJECTIVES: To study retrospectively CA 125, CD44, and epithelial membrane antigen (EMA) expression in renal cell carcinoma and their role as prognostic factors. CD44 is a cell adhesion molecule, and CA 125 and EMA are tumor associated antigens used in the diagnosis and monitoring of the outcome and response to treatment of various human malignancies. Their expression and prognostic significance after resection of renal cell carcinoma have not been adequately studied. METHODS: The expression of CA 125, CD44, and EMA were studied immunohistochemically and correlated with the outcome of 92 patients who underwent nephrectomy for renal cell carcinoma. RESULTS: Positive staining was found for CA 125 in 28 patients (30.43%), CD44 in 48 patients (52.17%), and EMA in 74 patients (80.43%). CA 125 expression was increased in those with higher T stage (P <0.001) and histologic grade (P = 0.007). An inverse relationship was found between EMA expression and grade (P <0.001). The median follow-up was 41.5 months (range 30 to 65). The median survival for positive and negative patients was 34.6 versus 54.3 months for CA 125 (P = 0.0044), 48.3 versus 51.5 months for CD44 (P = 0.4677), and 53.2 versus 34 months for EMA (P = 0.0046). Multivariate analysis showed that CA 125 and EMA expression were independent prognostic factors (P = 0.021 and P = 0.018, respectively). Subgroup analysis showed that CA 125 expression predicted a significantly higher probability of death (28.6% versus 8%, P =0.0413) in patients with T1 or T2 tumors. CONCLUSIONS: CA 125 and EMA appear to be useful prognostic markers in renal cell carcinoma. Additional studies are needed to determine the value of these markers as a means of selection for postoperative management. PMID- 12893367 TI - Vascular endothelial growth factor gene-460 C/T polymorphism is a biomarker for prostate cancer. AB - OBJECTIVES: To use the Bst U I polymorphism as a genetic marker in the search for the association between patients with prostate cancer and normal control subjects. The formation or progression of prostate cancer is presumed to be associated with a polymorphism of the vascular endothelial growth factor (VEGF) gene. The most frequently seen polymorphism is Bst U I (C to T) located at the 460th nucleotide upstream of the VEGF gene. METHODS: A normal control group of 119 healthy people and 96 patients with prostate cancer were examined. The polymorphism was seen after polymerase chain reaction-based restriction analysis. RESULTS: The analysis revealed significant differences between normal individuals and patients with cancer (P <0.001). Also, the distribution of the "TT" homozygote in the patient group was greater than that in the control group. The odds ratio per copy of the "T" allele was 2.3 (95% confidence interval 1.4 to 3.8) and was 2.2 (95% CI 1.3 to 3.8) when adjusted for age. No statistically significant differences in clinical stage or grade were found. We also categorized the 54 patients who received hormonal therapy into response and nonresponse groups, but no statistically significant differences between these two groups were revealed (P = 0.110, Fisher's exact test). CONCLUSIONS: The Bst U I polymorphism of the VEGF gene is a suitable genetic marker of prostate cancer but cannot be used in the prediction of the outcome of patients who have received hormonal therapy. PMID- 12893368 TI - Clinical significance of allelotype profiling for urothelial carcinoma. AB - OBJECTIVES: To perform a global loss of heterozygosity (LOH) analysis on a cohort of urothelial carcinoma to investigate the clinical implication of specific chromosomal loss. Allelic deletions detected as LOH have been used to study the markers for carcinogenesis. METHODS: We examined the allelic loss on 14 chromosomal regions in a total of 71 cases of urothelial carcinoma. The results were analyzed in relation to biologic indicators of urothelial carcinoma and the clinical outcome with a mean follow-up of 101 months. RESULTS: The incidence of LOH in order of frequency was 9p (54.9%), 9q (49.3%), 13q (40.8%), 14q (40.8%), 10q (39.4%), 17p (39.4%), 8p (38.0%), 21q (36.6%), 11p (31.0%), 18q (23.9%), 4q (21.1%), 3p (16.9%), 6q (14.1%), and 1q (8.5%). Positive association with one of the indicators was observed in 3p, 9p, 9q, 10q, 14q, and 18q. The chromosomes that correlated with two biologic indicators were 4q, 6q, 11p, 17p, and 21q. Univariate analysis found that patients having combined 9p and 14q deleted tumors had particularly poor long-term survival compared with those with other patterns of chromosomal alterations (P = 0.01). In the multivariate model, nonpapillary tumors had a greater risk of recurrence, and stage classification was the only important indicator in predicting patient survival (P = 0.04). CONCLUSIONS: LOH assessment does not provide independent prognostic value compared with stage classification. However, chromosomes 4q, 6q, 9p, 11p, 14q, 17p, and 21q may harbor important tumor suppressor genes involved in the progression of urothelial carcinogenesis. PMID- 12893369 TI - Moses Swick, the father of intravenous urography. PMID- 12893370 TI - Recurrent bleeding from intercostal arterial pseudoaneurysm after retroperitoneal laparoscopic radical nephrectomy. PMID- 12893371 TI - [Checking and maintenance of anaesthetic machines for improved safety in anaesthesia]. PMID- 12893372 TI - [Survey and budgetary impact of anaesthesia machines failures]. AB - OBJECTIVE: To assess the incidence and the causes of failures of anaesthesia machines in relation to aging. Study design. - Prospective survey from january 1996 to july 2000. MATERIAL AND METHODS: The causes (mechanical or electronic), the moment of identification (checklist, maintenance operation or quality-control operation) of each anaesthetic machine failure, the repair cost and the maintenance cost of 14 anaesthetic machines have been collected and entered into a database. RESULTS: Over 31,948 anaesthesia delivered during the period of the study, 614 failures have been declared: 53% were related both to mechanical problems or monitoring failure and 40% were identified during the pre-operative checklist. In half of the cases, a specially trained anaesthetic nurse was able to correct the failure in the operating theatre. The annual rate of anaesthetic machine failure remained stable over the study period and the annual maintenance cost is approximatively 10% of the initial machine value. No procedure was cancelled because of a machine technical failure. CONCLUSION: Anaesthetic machine failure rate change according to the time should not be criteria for remplacement if rigorous quality control and maintenance operation are used. PMID- 12893373 TI - [Pseudomonas aeruginosa epidemiology in intensive care units: importance of cross transmission]. AB - OBJECTIVES: To update the local epidemiological data of Pseudomonas aeruginosa in intensive care units (ICU) by assessing the colonisation incidence rate and the level of cross-transmission. METHODS: Study carried out in both adult ICUs of the university-hospital of Besancon during a 2 years period. Clinical and surveillance specimens were screened for P. aeruginosa. Pulsed-field-gel electrophoresis was used as genotyping method to evaluate the rate of cross transmission. RESULTS: During the study, 314 patients were positive for P. aeruginosa (incidence rate of 19.1 patients per 100 admitted patients). One hundred sixty-six of these patients were detected with a clinical specimen and 148 with a screening specimen. Seventy-seven patients were colonised upon admission in the intensive care unit and 237, negative on admission, became positive during their stay. Of the ICU-acquired cases, the mean length of stay before P. aeruginosa colonisation was acquired was 15.7 days. Genotyping revealed that 53.5% of P. aeruginosa colonisation was acquired via cross-transmission (respectively 48.1% in the medical ICU and 59.2% in the surgical ICU); the other cases probably originated from endogenous sources. CONCLUSION: The incidences of P. aeruginosa colonisation upon admission and during hospitalisation are consistent with other french and european studies. Although we probably over estimated the rate of cross-transmission, our results demonstrate that cross transmission may be a major cause of P. aeruginosa dissemination in ICUs. PMID- 12893374 TI - [Neostigmine added to bupivacaine in axillary plexus block: which benefit?]. AB - INTRODUCTION: Recent study showed that neostigmine (500 microg) by intra articular produces postoperative analgesia without adverse effect. The author's goal was to determine whether 500 microg of neostigmine added to bupivacaine in axillary plexus block could prolonged postoperative analgesia without increasing the incidence of side effects. METHODS: Ninety patients scheduled for orthopedic or plastic surgery with axillary plexus block were randomly assigned to one of 3 groups : group 1 (TGr n = 30) received saline solution (1 ml) in the axillary plexus, group 2 (NAGr n = 30) received 500 microg (1 ml) neostigmine in the axillary plexus and group 3.500 microg neostigmine subcutaneously (NSGr n = 30). We evaluated visual analog pain scores (VAS), the consumption of the ketoprofene, nausea and vomiting incidence during the first 24 h. ANOVA, Kruskall Wallis and Fisher tests were used for statistical analysis. A p value of <0.05 was considered significant. RESULTS: The VAS score was lower in NAGr (21 +/- 18) vs NSGr (31 +/- 14) and control group TGr (45 +/- 2) (p < 0.05). The consumption of the ketoprofene is 127 +/- 65 mg in NAGr vs 150 +/- 53 mg in NSGr and 200 +/- 50 mg in group TGr (p = 0.02). Incidence of nausea and vomiting was 3.5% in NAGr vs 6.8% in NSGr and 0% for TGr. CONCLUSION: Neostigmine combined to a mixture of lidocaine and bupivacaine prolongs postoperative analgesia after axillary plexus block. PMID- 12893375 TI - [Aetiologies of lithium overdose: 10-year experience of Marseille poison centre]. AB - OBJECTIVE: Lithium is used for control of bipolar disorders. In order to precise the different circumstances at the origin of poisonings, the authors present the cases of lithium intoxication observed in the Marseille poison centre between January 1991 and December 2000. STUDY DESIGN: Retrospective study. METHODS: Three hundred and four cases were observed during the studied period (1 patient a case), concerning 6 different circumstances. For 3 of them, the symptoms were mild: accidental ingestion with children (13 cases); mistakes on the quantities of ingested tablets (43 cases); elevation of lithium blood level due to diuretic therapy (8 cases). For 2 other circumstances, the clinical signs were more severe: treated patients who developed renal failure (15 cases, 6 patients managed in intensive care unit [ICU], 1 death) or dehydration (35 cases, 8 patients treated in ICU and 1 death). Finally, the most severe cases were collected with suicide attempts. Fifty-six percent of the patients were managed in ICU, 5% needed haemodialysis, 10% had cardiac (repolarization disturbances) or neurological (seizures) complications, 2% died. CONCLUSION: The severity of lithium poisonings depends of the circumstances. Ingestion of high quantities of sustained released tablets is the most dangerous situation. Accidental ingestion, even with children, must be considered as less severe situations. PMID- 12893376 TI - [Epidemiology of infections due to Pseudomonas aeruginosa]. PMID- 12893377 TI - [Pseudomonas aeruginosa: a model of choice for the study of opportunistic pathogen]. PMID- 12893378 TI - [Mechanisms of resistance to betalactam antibiotics in Pseudomonas aeruginosa]. PMID- 12893379 TI - [Pseudomonas aeruginosa: from colonisation to infection]. PMID- 12893380 TI - [Therapeutic strategies for infections due to Pseudomonas aeruginosa]. PMID- 12893381 TI - [Unusual clinical aspects of infections due to Pseudomonas aeruginosa]. PMID- 12893382 TI - [Management of an outbreak of infections due to Pseudomonas aeruginosa]. PMID- 12893383 TI - [General anaesthesia for Caesarean section in a patient with pulmonary atresia and an intact intraventricular septum]. AB - We report the case of a 22-year-old primigravida patient presenting pulmonary atresia with intact ventricular septum, a rare congenital heart disease. Caesarean delivery was performed at 32 weeks of gestation because of moderate maternal function deterioration and foetal growth restriction. General anaesthesia was used because of the presence of lumbar Harrington rods. The administration of etomidate, celocurine and a continuous perfusion of remifentanil right from induction ensured haemodynamic stability and a rapid emergence and recovery. PMID- 12893384 TI - [Peroperative desaturation during gynaecological laparoscopy-hysteroscopy: an unknown aetiology, pleural extravasation]. AB - A 41-year-old patient presented several episodes of desaturation during a gynaecological laparoscopy. The major complication of this procedure is the venous air embolism. Several other side-effects have been reported: heart rate disorders, subcutaneous emphysema or pneumothorax. Pleural effusions during gynaecologic laparoscopy are apparently rare and the volume of effusion must be important to induce clinical symptoms. This fact can probably explain the frequent difficulty of diagnosis. The role of the diaphragmatic lymphatic network and other physiologic aspects are discussed in this article. PMID- 12893385 TI - [Acute cupric sulfate intoxication: pathophysiology and therapy about a case report]. AB - A case of acute copper sulfate intoxication is presented here, as an illustration of high toxic copper dose. A 38-years-old patient with a light mental deficit ingested half a glass of copper sulfate. This patient first suffered from nausea and vomiting, then an intravascular haemolysis occurred during the hospitalisation in our intensive care unit. The outcome was favourable under aetiologic and symptomatic treatment: administration of D-penicillamine as a copper antidote and packed red cell transfusion. The patient left the intensive care unit after 9 days. Acute copper intoxication is not an exceptional situation, with a potentially severe outcome. PMID- 12893386 TI - [Cranial subdural haematoma following spinal anaesthesia]. PMID- 12893387 TI - [Peripheral nerve block of the upper limb: which dose and which concentration should be use?]. PMID- 12893388 TI - [Can a preliminary survey help to elaborate information to patient sheet?]. PMID- 12893389 TI - [The ideal length of catheter into lumbar epidural space]. PMID- 12893391 TI - [Peripheral nerve block of limbs in the adult]. PMID- 12893392 TI - C-reactive protein and coronary events following percutaneous coronary angioplasty. AB - PURPOSE: We investigated the associations between baseline C-reactive protein levels in patients undergoing percutaneous coronary angioplasty and death, nonfatal myocardial infarction, and repeat revascularization during 14 months of follow-up. METHODS: In a single-center, prospective, cohort study, plasma levels of C-reactive protein were measured in 1458 consecutive patients undergoing elective or urgent coronary angioplasty. Patients were followed at 12 to 14 months for the occurrence of death, nonfatal myocardial infarction, and repeat revascularization. RESULTS: The incidence of death or myocardial infarction was 6.1% (44/716) in patients with an increased C-reactive protein level (>3 mg/L) and 1.5% (11/742) in patients with a normal level (relative risk [RR] = 4.4; 95% confidence interval [CI]: 2.2 to 8.5; P <0.0001). In a multivariate logistic regression model, an increased C-reactive protein level was an independent predictor of death or nonfatal myocardial infarction (RR = 3.6; 95% CI: 1.8 to 7.2; P =0.0001). The incidence of repeat revascularization was similar in patients with or without an increased C-reactive protein level (23% [168/716] vs. 22% [163/742], P = 0.54). Statin therapy at the time of the procedure was associated with a lower mean (+/- SD) C-reactive protein level (5.8 +/- 9.7 mg/L vs. 7.2 +/- 12.1 mg/L, P =0.02), but was not associated with the risk of death, nonfatal myocardial infarction, and repeat revascularization during follow-up. CONCLUSION: An increased C-reactive protein level is an independent prognostic indicator for the occurrence of death or nonfatal myocardial infarction following coronary angioplasty, but is not associated with the need for repeat revascularization. PMID- 12893393 TI - Long-term follow-up of patients with culture-confirmed Lyme disease. AB - PURPOSE: To determine the long-term outcome of patients with culture-confirmed Lyme disease. METHODS: We analyzed data collected prospectively on adult patients from a highly endemic area in New York State who were diagnosed with early Lyme disease between 1991 and 1994. Patients with culture-confirmed erythema migrans were evaluated at baseline, 7 to 10 days, 21 to 28 days, 3 months, 6 months, 1 year, and annually thereafter. All patients were treated with antibiotics at the time of diagnosis. RESULTS: We evaluated 96 cases on 709 separate occasions (median, eight evaluations per case). The erythema migrans rash resolved within 3 weeks in all of the 94 evaluable cases, none of whom developed an objective extracutaneous manifestation of Lyme disease. Of the 81 cases who were followed for >/=1 year, all but 8 (10%) were asymptomatic at their last visit, a mean (+/- SD) of 5.6 +/- 2.6 years into follow-up, and only 3 (4%) were symptomatic at every follow-up visit. Intercurrent tick bites were reported by 45 cases (47%), and 14 (15%) developed a second episode of erythema migrans. Four other cases who were asymptomatic seroconverted between years 2 and 5. CONCLUSION: The long-term outcome of patients with erythema migrans after antibiotic therapy was excellent, but patients from a highly endemic area in New York State remained at high risk of re-exposure to ticks and reinfection. Subjective symptoms during follow-up evaluations tended to be mild to moderate, intermittent, and associated with more symptomatic illness at the time of initial diagnosis. PMID- 12893394 TI - The effects of cognitive impairment on mortality among hospitalized patients with heart failure. AB - PURPOSE: Cognitive impairment is a common, potentially reversible condition among older patients with heart failure. Because cerebral metabolic abnormalities have been associated with reduced survival in younger patients with advanced heart failure, we assessed the effect of cognitive impairment on the survival of older patients with heart failure. METHODS: The association between cognitive dysfunction and in-hospital mortality was assessed in 1113 patients (mean [+/- SD] age, 78 +/- 9 years) who had been admitted for heart failure to 81 hospitals throughout Italy. One-year mortality was assessed in 968 patients with heart failure (age, 76 +/- 10 years) participating in the same study. Cognitive impairment was defined as a Hodkinson Abbreviated Mental Test score <7. RESULTS: In-hospital death occurred in 65 (18%) of the 357 participants with cognitive impairment and in 26 (3%) of the 756 patients with normal cognition (P <0.0001). Out-of-hospital mortality was 27% (51/191) among patients with cognitive impairment and 15% (115/777) among other participants (P <0.0001). In multivariate Cox regression models, decreasing levels of cognitive functioning were associated with increasing in-hospital mortality; cognitive impairment was associated with an almost fivefold increase in mortality (relative risk = 4.9; 95% confidence interval: 2.9 to 8.3) after adjusting for several potential confounders. CONCLUSION: Cognitive impairment is an independent prognostic marker in older patients with heart failure. Assessment of cognitive functioning, even by simple screening tests, should be part of the routine assessment of elderly patients with heart failure. PMID- 12893395 TI - Anemia and decline in physical performance among older persons. AB - PURPOSE: Anemia is prevalent in old age and is potentially modifiable, but its effects on physical function have not been determined. We examined whether anemia in older persons increases the risk of subsequent decline in physical function, as measured by objective performance-based tests. METHODS: Participants in this 4 year prospective cohort study included 1146 participants, aged 71 years or older, living in Iowa and Washington counties, Iowa. Anemia was defined according to World Health Organization (WHO) criteria as a hemoglobin concentration below 12 g/dL in women and below 13 g/dL in men. An assessment of standing balance, a timed 2.4-m walk, and a timed test of five chair rises were used to assess physical performance; these were combined into a 0 (poor) to 12 (excellent) summary scale. RESULTS: After adjustment for baseline performance score, health status, and demographic characteristics, anemia was associated with greater mean decline in physical performance over 4 years; the adjusted mean decline was 2.3 (95% confidence interval [CI]: 1.7 to 2.8) in subjects with anemia and 1.4 (95% CI: 1.2 to 1.5) in those without anemia (P = 0.003). The association between anemia and greater physical decline was also present in participants who were free of diseases associated with anemia (cancer, infectious disease, and renal failure), and after adjustment for serum cholesterol, iron, and albumin levels. Persons with borderline anemia, a hemoglobin concentration within 1 g/dL above the WHO criteria, also showed greater mean physical decline (1.8; 95% CI: 1.5 to 2.2) than did those with higher hemoglobin concentrations (P = 0.02). CONCLUSION: This study suggests that anemia in old age is an independent risk factor for decline in physical performance. PMID- 12893396 TI - Fecal occult blood testing in a general medical clinic: comparison between guaiac based and immunochemical-based tests. AB - PURPOSE: Guaiac-based fecal occult blood tests are limited by poor patient compliance, and low sensitivity, specificity, and positive predictive value. Newer immunochemical-based tests are designed to improve accuracy and patient compliance. We compared patient compliance and the test characteristics of these two types of tests. METHODS: The laboratory outcomes associated with use of different fecal occult blood tests were examined in a Veterans Affairs-based general medicine clinic that was divided into two firms with similar patient and provider characteristics. Tests were ordered for colorectal cancer screening or for symptom evaluation. Patients were given one of the two tests depending on their firm. The completion and positivity rates, time to test completion, completion of diagnostic follow-up, and positive predictive values were compared. RESULTS: The percentage of returned test cards was similar between the two groups (47% [1369/2964] for guaiac-based tests vs. 48% [1410/2965] for immunochemical based tests) as was the positivity rate (9.0% [122/1396] and [128/1410] for both groups). In patients with positive tests who underwent further colon evaluation, the proportion with adenomas was similar between groups (59% [38/64] vs. 58% [40/69]). However, 17% (12/69) with a positive immunochemical-based test had an adenoma >1 cm or a colorectal malignancy, versus 30% (19/64) for guaiac-based tests (P = 0.09). CONCLUSION: Overall, immunochemical-based and guaiac-based fecal occult blood tests had comparable performance. However, although immunochemical-based testing is reported to be easier for patients than guaiac based testing, we found that patients were no more likely to return cards for analysis. The similar positive predictive value and additional cost of immunochemical-based tests call into question their utility in general practice. PMID- 12893397 TI - Parathyroid hormone-related peptide and survival of patients with cancer and hypercalcemia. AB - PURPOSE: Parathyroid hormone-related peptide (PTHrP) is the predominant cause of malignancy-associated hypercalcemia. However, its prognostic utility is unclear. We aimed to determine the prognostic value of serum PTHrP levels in patients who had hypercalcemia associated with malignancy. METHODS: In this prospective case series, we evaluated 76 patients with a diagnosis of cancer and hypercalcemia (serum calcium level >/=10.3 mg/dL on at least two occasions). PTHrP levels >/=1 pmol/L were considered elevated. We used multivariate Cox regression analysis to identify factors associated with mortality. RESULTS: Fifty patients (66%) died during follow-up. In a multivariate analysis, higher pretreatment calcium levels and elevated PTHrP levels were associated with increased mortality, with effects of PTHrP varying by age (P = 0.03). Survival was associated with pretreatment calcium levels both in patients over 65 years of age (hazard ratio [HR] per mg/dL = 1.5; 95% confidence interval [CI]: 1.2 to 1.8; P <0.001) and in patients aged 65 years or less (HR = 1.3; 95% CI: 1.1 to 1.5; P = 0.003). Adjusted for pretreatment calcium levels, elevated PTHrP levels were associated with increased mortality in patients aged 20 ms. PMID- 12893423 TI - Alkaloid fraction of Uncaria rhynchophylla protects against N-methyl-D-aspartate induced apoptosis in rat hippocampal slices. AB - Uncaria rhynchophylla is a medicinal herb which has sedative and anticonvulsive effects and has been applied in the treatment of epilepsy in Oriental medicine. In this study, the effect of alkaloid fraction of U. rhynchophylla against N methyl-D-aspartate (NMDA)-induced neuronal cell death was investigated. Pretreatment with an alkaloid fraction of U. rhynchophylla for 1 h decreased the degree of neuronal damage induced by NMDA exposure in cultured hippocampal slices and also inhibited NMDA-induced enhanced expressions of apoptosis-related genes such as c-jun, p53, and bax. In the present study, the alkaloid fraction of U. rhynchophylla was shown to have a protective property against NMDA-induced cytotoxicity by suppressing the NMDA-induced apoptosis in rat hippocampal slices. PMID- 12893425 TI - Metastasis suppressor pathways--an evolving paradigm. AB - A greater understanding of the processes of tumor invasion and metastasis, the principal cause of death in cancer patients, is essential to determine newer therapeutic targets. Metastasis suppressor genes, by definition, suppress metastasis without affecting tumorigenicity and, hence, present attractive targets as prognostic or therapeutic markers. This short review focuses on those twelve metastasis suppressor genes for which functional data exist. We also outline newly identified genes that bear promising traits of having metastasis suppressor activity, but for which functional data have not been completed. We also summarize the biochemical mechanism(s) of action (where known), and present a working model assembling potential metastasis suppression pathways. PMID- 12893426 TI - Effect of PSC 833, a potent inhibitor of P-glycoprotein, on the growth of astrocytoma cells in vitro. AB - Malignant astrocytomas have been found to express P-glycoprotein (Pgp, mdr1 gene product). It was hypothesized that in addition to conferring multidrug resistance, Pgp is intimately associated with the development of astrocytomas. Accordingly, we studied the effect of PSC 833 (PSC, Novartis), a potent inhibitor of Pgp, on the growth of Pgp-expressing astrocytoma cells. The results showed that in all the cell lines tested, PSC (10-60 microM) inhibited the growth as well as induced cell death. Cells exposed to PSC exhibited DNA ladder characteristic of apoptosis. PSC-induced cell death could be reversed by Z-VAD fmk, a general caspase inhibitor, indicating that PSC-induced cell death was characteristic of caspase-mediated apoptosis. These results suggest a novel therapeutic strategy in the treatment of malignant astrocytomas by inhibitors of Pgp. PMID- 12893427 TI - Beta-catenin mutations in liver tumors induced by 2-amino-3,4-dimethylimidazo[4,5 f]quinoline in CDF1 mice. AB - Heterocyclic amines are potent mutagens and carcinogens formed in cooked protein rich foods. In this study, we screened liver tumors induced by 2-amino-3,4 dimethylimidazo[4,5-f]quinoline (MeIQ) in CDF1 mice for beta-catenin and APC mutations and other genetic alterations shown to occur in human hepatocellular carcinomas (HCC), including mutations in the p53 and H-ras genes, c-myc amplification and E-cadherin promoter methylation. SSCP followed by direct DNA sequencing revealed mutations in exon 2 of the beta-catenin gene in 2 of 16 liver tumors (12.5%). Promoter methylation of the E-cadherin gene was detected in one liver tumor induced by MeIQ. There were no mutations in the mutation cluster region of the APC gene, in exons 5-8 of the p53 gene, or in codons 12, 13 and 61 of the H-ras gene, nor c-myc amplification in any of liver tumors induced by MeIQ. These data indicate that except for the occasional disruption of the Wnt pathway through beta-catenin mutations, the genetic pathways involved in the development of HCC differ significantly between human liver cancer and tumors induced in mice by MeIQ, but do not rule out the possibility that heterocyclic amines constitute a carcinogenic risk factor in humans. PMID- 12893428 TI - Anticarcinogenic activity of natural sweeteners, cucurbitane glycosides, from Momordica grosvenori. AB - To search for cancer chemopreventive agents from natural resources, many phytochemicals and food additives have been screened. Consequently, two natural sweeteners, mogroside V and 11-oxo-mogroside V isolated from the fruits of Momordica grosvenori, exhibited strong inhibitory effect on the primary screening test indicated by the induction of Epstein-Barr virus early antigen (EBV-EA) by a tumor promoter, 12-O-tetradecanoylphorbol-13-acetate (TPA). These sweet glycosides, having cucurbitane triterpenoid aglycon, exhibited the significant inhibitory effects on the two-stage carcinogenesis test of mouse skin tumors induced by peroxynitrite (ONOO-) as an initiator and TPA as a promoter. Further, 11-oxo-mogroside V also exhibited the remarkable inhibitory effect on two-stage carcinogenesis test of mouse skin tumor induced by 7,12-dimethylbenz[a]anthracene (DMBA) as an initiator and TPA as a promoter. PMID- 12893429 TI - Whey proteins protect more than red meat against azoxymethane induced ACF in Wistar rats. AB - Protein type and density have been shown to influence colon cancer risk using a carcinogen-induced rat model. It is suggested that red meat may promote colon cancer risk more than whey proteins. The aim of this study was to evaluate the influence of red meat, whey protein and their density in the diet on the number of aberrant crypt foci (ACF), preneoplastic markers in Wistar rats. The sources of protein, red meat as barbecued kangaroo muscle meat, and whey protein concentrate were fed to rats to provide 8, 16 and 32% protein by weight in a modified AIN-93 diet with low fiber, low calcium and high polyunsaturated fat. Adult Wistar rats (13 weeks of age) were fed these diets for 4 weeks and then two s.c. injections of azoxymethane, 15 mg/kg BW, were administered 1 week apart. Diets were fed for a further 8 weeks, rats were then killed, their colons fixed in formalin saline and stained with methylene blue to quantify ACF number. Fecal samples were collected and the fecal water was isolated for quantification of heme and thiobarbituric acid reactive substances. Increasing red meat density correlated positively, while increasing dairy protein density correlated negatively with rate of weight gain (p<0.05). Dietary intake was not significantly affected by protein type or density. The 32% whey protein group had significantly less ACF in the proximal colon in comparison to the 16 and 32% red meat groups (p<0.05). This reduction in ACF number in the whey protein group may be caused by hormones associated with the reduction in weight gain, and/or by components of whey protein concentrate such as cysteine, lactose and conjugated linoleic acid which have been shown to have anti-cancer effects. Using ACF number as an index, whey protein appeared to be more protective than red meat. PMID- 12893430 TI - Topical inhibition of oral carcinoma cell with polymer delivered celecoxib. AB - Celecoxib has a potential role for oral cancer chemoprevention but its systemic side effects are a concern. Topical chemoprevention is a promising way to reduce the toxicity. This study was designed to determine whether topical application of polymer delivered celecoxib would have an inhibitory effect on human oral carcinoma cells. Seventeen nude mice were intradermally inoculated with the carcinoma cells, and then were divided into control and treatment groups. Tumor growth was measured for 15 days, at which significant difference was found between two groups (P<0.001). This study indicates a potential role of polymer film delivered celecoxib for topical inhibition of oral cancer. PMID- 12893431 TI - The selective in vitro cytotoxicity of carcinoma cells by AZT is enhanced by concurrent treatment with delocalized lipophilic cations. AB - This study assessed the selective growth inhibitory effect on cultured carcinoma cells by 3'-azido-3'-deoxythymidine (AZT), as a single agent, and in combination with delocalized lipophilic cations (DLCs) that are known to inhibit mitochondrial function. In cytotoxicity assays, treatment of cells with varying concentrations of AZT induced a dose-dependent inhibition of cell growth of the human carcinoma lines DU-145 (prostate; IC50 at 24 microM), MCF-7 (breast; IC50 at 22 microM), and CX-1 (colon; IC50 at 23 microM), yet caused no significant effect on the growth of the control epithelial cell line CV-1 (monkey kidney) at a concentration as high as 50 microM AZT. Combination treatment employing a constant concentration (1.25 microM) of the DLC dequalinium chloride (DECA) plus varying concentrations of AZT (0-50 microM) enhanced the AZT-induced cytotoxicity of carcinoma cells at least fourfold for MCF-7 and CX-1 cells (IC50 at 5 microM AZT), and twofold for DU-145 cells (IC50 at 11 microM AZT). Similar results were obtained in DU-145 cells using a constant concentration of the DLC MKT-077 (1.0 microM) and varying concentrations of AZT (IC50 at 12.5 microM). As expected, the drug combination of constant DLC and varying AZT had no significant effect on the growth of CV-1 cells. Clonogenic assays demonstrated up to 20-fold enhancement of selective carcinoma cell killing by combination vs. single agent treatment, depending on the specific drug combination and concentrations used. It is hypothesized that the efficacy of the AZT/DLC drug combination in carcinoma cell killing may be based on a dual selectivity involving inhibition of mitochondrial energy metabolism and inhibition of DNA synthesis due to limited deoxythymidine monophosphate availability. PMID- 12893432 TI - The p53 codon 72 polymorphism in Thai nasopharyngeal carcinoma. AB - Nasopharyngeal carcinoma (NPC) is a serious cancer in Thailand. To address on the genetic risk factor for NPC, we investigated association between the p53 codon 72 polymorphism (Pro/Arg) and NPC susceptibility in the Thai. The genotype frequency of this polymorphism in 102 NPC patients and 148 age-matched healthy controls was determined by using a PCR-RFLP assay. No statistically significant difference in the overall genotype frequencies or allele frequencies between cases and controls was observed. Among NPC patients, no statistical significant difference in p53 genotype frequencies between sex, histological types and clinical stages was observed. When cases and controls were categorized into 3 groups of age, >40, >45 and >50 years, the frequencies of p53 genotype in the cases were significantly different from those of the controls (P<0.05). The p53 Pro homozygotes with age of >40 years had twofold to threefold higher risk to develop NPC and the risk was increased with age (Odds Ratio (OR) increased from 2.01 to 2.63 by age >40 to >50 years). Individuals of age >40, >45 and >50 years with an Arg/Arg genotype and of age >45 and >50 years with combined Arg/Arg or Arg/Pro genotype had lower risk to develop NPC than those with Pro/Pro genotype (P<0.05). Collectively, we suggest that the p53 gene polymorphism may associate with NPC susceptibility in Thai population, particularly the Pro/Pro genotype carriers with age of >40 years. The detection of this p53 polymorphism may be a useful tool for screening of early stage and diagnosis of NPC. PMID- 12893433 TI - Human herpes viruses in non-melanoma skin cancers. AB - We examined the possible involvement of human herpes viruses in sporadic non melanoma skin cancer of Greek patients. Polymerase chain reaction (PCR) based detection assays were utilized for the detection of viral cytomegalovirus (CMV), herpes simplex virus (HSV) and Epstein-Barr virus (EBV) genomes in 24 squamous cell carcinomas (SCC), five Bowen's disease, 72 basal cell carcinomas (BCC) specimens and eight premalignant lesions. Forty-two of 109 (38.5%) skin lesions were found positive for CMV DNA. The highest incidence was 6/8 (75%) observed in specimens with premalignant lesions. The incidence was 37.5% (27/72) in BCC, 33% (8/24) in SCC and 20% (1/5) in extragenital Bowen's disease. All samples were negative for HSV-1/2 and EBV DNA as assessed by our PCR based assay. The CMV infection showed no statistically significant correlation with the histological type, age, site of lesion or sex. Our results give a strong indication of the possible involvement of CMV in non-melanoma skin cancer development. PMID- 12893434 TI - Circulating numbers of endothelial progenitor cells in patients with gastric and breast cancer. AB - Angiogenic factors like VEGF or G-CSF were reported to mobilize endothelial progenitor cells (EPCs) from the bone marrow. These EPCs were shown to be incorporated in the neovessels of developing tumors. Although the concentrations of angiogenic factors in the peripheral blood were reported to be elevated in cancer patients, the number of circulating EPCs has not been previously investigated. In this study, the number of EPCs circulating in the blood in 16 healthy controls and 71 newly diagnosed cancer patients was examined by a culture assay of peripheral blood mononuclear cells. The number of circulating EPCs was not found to be increased in cancer patients, although the plasma levels of VEGF were elevated. It is suggested that VEGF, at concentrations typical of those observed in the blood of cancer patients, does not mobilize EPCs into the peripheral blood. PMID- 12893435 TI - Mutation of the class I beta-tubulin gene does not predict response to paclitaxel for breast cancer. AB - Mutation of the class I beta-tubulin gene has been reported to be one of the mechanisms that cause resistance to paclitaxel. To assess the relationship between paclitaxel-resistance and class I beta-tubulin gene mutation in breast cancer, Japanese patients with breast cancer were screened for the class I beta tubulin gene mutation. Total RNA was isolated from 82 breast cancer specimens and the corresponding normal tissues. Twenty-four of the 82 patients were treated with paclitaxel preoperatively and 12 of them did not respond to the treatment. Of the 82 breast cancer patients, 15 (18.3%) had silent polymorphism in exon 4, Leu217Leu (CTG/CTA). However, no mutations showing amino acid substitution of the beta-tubulin gene were detected in any of the patients, including 12 patients who did not respond to paclitaxel. Class I beta-tubulin gene mutation with amino acid substitution was not detected in 82 breast cancer specimens. Our results suggest that mutation of the class I beta-tubulin gene is unlikely to play an important role in the mechanism of resistance to paclitaxel in breast cancer. PMID- 12893436 TI - Estrogenic and antiestrogenic regulation of MMP-2 and MMP-13 mRNA in RUCA-I endometrial tumor cells in vitro and in vivo. AB - We investigated the influence of estrogenic and antiestrogenic treatment on proteolytic activity--especially on MMP-2 and MMP-13--in the RUCA-I transplantable endometrial tumor model. Morphological studies demonstrate that RUCA-I cells are forming highly differentiated gland-like structures by remodelling and invading the underlying ECM. Estrogens upregulate the mRNA levels of MMP-2 and MMP-13 in the rat uterus. Treatment with the pure antiestrogen ICI 182,780 results in the downregulation of MMP-2 and MMP-13 mRNA. The same regulation for MMP-13 mRNA is found in vitro in RUCA-I cells. In contrast, in the transplantation tumor, the mRNA level of MMP-13 is repressed by estrogens and induced by ICI 182,780. MMP-2 mRNA is not regulated by hormones in the transplantation tumor and in RUCA-I cells. The divergent regulation suggests a varying influence of cell-cell-, cell-extracellular matrix interactions and soluble factors. PMID- 12893437 TI - Osteopontin overproduced by tumor cells acts as a potent angiogenic factor contributing to tumor growth. AB - Angiogenesis, which is essential for tumor growth, is regulated by various angiogenic factors. Osteopontin (OPN) is expressed in various human tumors and is postulated to be involved in tumor progression. We have recently reported that culture medium with murine neuroblastoma C1300 cells transfected with OPN gene significantly stimulates human umbilical vein endothelial cell migration and induces neovascularization in mice by dorsal air sac assay. However, the effect of OPN on tumorigenesis as an angiogenic factor remains to be clarified. In this study, we injected the OPN-transfected C1300 cells and control cells into the nude mice subcutaneously. OPN-overexpressing C1300 cells significantly formed rapidly growing tumor as compared to the control cells in mice, although in vitro and in vivo cell growth rates were similar. In vivo tumorigenecity of these cells correlated with the amount of secreted OPN protein. In addition, neovascularization of OPN-transfected tumor was significantly increased in comparison with those of control cells by immunohistochemistry for CD31. In vitro chemoinvasiveness and gene expression of proteases including uPA, MMP2, 9, MT1 MMP, and cathepsin B, D, L, were not different between OPN-transfected and control cells determined with matrigel invasion assay and cDNA expression macroarray, respectively. Conclusively, these results strongly imply that OPN plays an important role in tumor growth through the enhancement of angiogenesis in vivo. PMID- 12893451 TI - Alcoholism and the dopaminergic system: a review. AB - Alcohol as well as other substances of abuse are reinforcing substances which manifest their effects through activation of the mesolimbic dopaminergic reward pathways of the brain. In animal genetic models of alcoholism, reduced dopamine levels and D2 dopamine receptor (DRD2) numbers have been found in the brains of alcohol-preferring animals. Dopamine receptor agonists reduce alcohol consumption, whereas antagonists, in general, show the opposite effect. Moreover, quantitative trait loci studies in animals suggest the DRD2 gene and the region proximate to this locus is a chromosomal "hot spot" for alcohol-related behaviors. Human studies provide additional support for connection between alcohol dependence and CNS dopaminergic function. In endocrinological studies, using dopamine receptor agonists, reduced dopaminergic activity has been found in more severe and more genetic types of alcoholics. Brain imaging studies are similarly revealing a diminished dopaminergic tone in alcoholics. Treatment of alcoholics with dopamine receptor agonists shows reduced alcohol consumption and improvements in other outcome measures. Molecular genetic studies in humans have identified an association of the Al allele of the DRD2 gene with alcoholism. Moreover, a diminished central dopaminergic function has been found in DRD2 A1 allele subjects using pharmacological, electrophysiological and neuropsychological studies. Further, treatment of alcoholics with a dopamine receptor agonist showed more salutary effects on alcoholics who carry than those who do not carry the DRD2 A1 allele. The A1 allele has also been associated with substance use disorders other than alcoholism, including and cocaine and nicotine dependence and polysubstance abuse. The emerging evidence suggests that the DRD2 is a reinforcement or reward gene. It could represent one of the most prominent single-gene determinants of susceptibility to severe substance abuse. However, the environment and other genes, when combined, still play the larger role. PMID- 12893452 TI - Alcohol dehydrogenases and aldehyde dehydrogenases among inbred strains of mice: multiplicity, development, genetic studies and metabolic roles. AB - Alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) are the major enzymes responsible for the metabolism of alcohols and aldehydes in the body. Both exist as a family of isozymes in mammals, and have been extensively studied in animal models, particularly among inbred strains of mice. Mouse ADH exists as at least three major classes, which are predominantly localized in liver (classes I and III), and in stomach/cornea (class IV). Mouse ALDH exhibits extensive multiplicity, several forms of which have been characterized, including ALDH1 (liver cytoplasmic/class 1 isozyme); ALDH2 (liver mitochondrial/class 2.); ALDH3 (stomach cytosolic/class 3); ALDH4 (liver microsomal/class 3); and ALDH5 (testis cytosolic/class 3). Biochemical, genetic and molecular genetic analyses have been performed on several of these enzymes, including studies on variant forms of ADH and ALDH. Distinct metabolic roles are proposed, based upon their tissue and subcellular distribution characteristics and the biochemical properties for these enzymes. PMID- 12893453 TI - Pathogenesis of alcoholic hepatic steatosis. AB - Chronic alcohol misuse is the most common cause of hepatic steatosis. The accumulation of lipid is reversible with abstinence, but some workers have suggested that the severity of hepatic steatosis predicts the progression with time to alcoholic cirrhosis. Triacylglycerol is the major accumulating lipid and subcellular fractionation and electron microscopy studies have shown accumulation of lipid droplets within the golgi fraction. This is consistent with the reports in both experimental animals and man of reduced hepatic secretion of very low density lipoprotein triacylglycerol which may be secondary to acetaldehyde induced disruption of the cytoskeletal elements. In addition, hepatic production of triacylglycerol increases, but most studies in animal models suggest that increased triacylglycerol synthesis becomes less important as hepatic lipid accumulates. PMID- 12893454 TI - The effects of acute administration of ethanol on jejunal protein synthesis and circulating insulin-like growth factor (IGF)-1 and IGF binding proteins in ad libitum fed and nutritionally restricted rats. AB - The effects of acute ethanol administration (75 mmol/kg body weight) to male Wistar rats (either ad libitum fed or nutritionally restricted) on fractional rates of protein synthesis in the jejunum was assessed together with the changes in IGF-1 and IGF binding protein concentrations. Acute administration of ethanol resulted in significant decreases in fractional rates of protein synthesis in the whole jejunum and jejunal seromuscular layers of both the ad libitum fed and nutritionally restricted animals. The synthesis rate per unit RNA (k(RNA), mg protein/day/mg RNA) in whole jejunum was reduced by 29% and 24% in the nutritionally restricted and ad libitum fed animals, respectively. Mean IGF-1 levels were lower in the nutritionally restricted group (871 +/- 36.9 microg/l) than the ad libitum animals (960 +/- 27.3 microg/l) although this did not reach significance. In contrast, administration of alcohol to both groups markedly reduced circulating IGF-1 levels (ad libitum: 518 +/- 19.8 microg/l, nutritionally restricted: 417 +/- 33.7 microg/l). Furthermore, ethanol treatment resulted in a three-fold increase in the intensity of a 30 kDa IGF binding protein (IGFBP) in the ad libitum fed animals and a fourfold increase in both 30 and 32 kDa IGFBP bands in the nutritionally restricted group as visualized by Western ligand blotting. Decreases in levels of IGF-1 allied with increased circulating small molecular weight IGFBPs may contribute to the reduction in fractional rates of protein synthesis in the gastrointestinal tract of ethanol treated rats. PMID- 12893455 TI - Acetaldehyde metabolism by liver mitochondrial ALDH from UChA and UChB rats: effect of inhibitors. AB - We have observed that blood acetaldehyde (AcH) levels after an ethanol dose were significantly higher in disulfiram-pre-treated UChA (low ethanol consumer) than in UChB (high ethanol consumer) rats. In order to explore these results further, we studied the effect of disulfiram (300 mg/kg i.p.) and chlorpropamide (80) mg/kg i.p.) pre-treatment on blood AcH levels after oral ethanol (60 mmol/kg) and on AcH metabolism by liver mitochondrial aldehyde(s) dehydrogenase(s) from UChA and UChB rats. AcH metabolism by liver mitochondrial aldehyde dehydrogenase (ALDH) was studied by following AcH disappearance rate and the formation of NADH at 340 nm in the incubation medium. The results showed that chlorpropamide, like disulfiram, produced a higher blood AcH level consistent with a greater inhibition of the low-Km mitochondrial ALDH in the UChA rats than in the UChB rats. These drugs did not inhibit the high Km mitochondrial ALDH. Kinetic studies of mitochondrial ALDH show that low-Km mitochondrial ALDH from UChB rats exhibits a higher affinity for NAD than UChA rats. This observation could explain the different inhibition of ALDH by both drugs, assuming that the inhibitors reduce NAD availability, the rate limiting step in the mitochondrial ALDH oxidation. PMID- 12893456 TI - Salivary glands in alcoholism: a histologic and sialographic study. AB - A qualitative and quantitative histological analysis of minor salivary glands was carried out in 60 alcoholics (20 alcoholics with liver damage, 20 without liver damage, 20 young alcoholics) and 20 matched control patients. A sialographic study was carried out in the same 60 alcoholic patients and in 40 non-alcoholic control individuals. Minor salivary gland biopsies were obtained and samples were processed for light microscopy. A quantitative morphometrical assessment was carried out by counting the number of acinar cells with a squared grid eye-piece. Both experimental and control samples presented very similar results demonstrating that, at least in this study, minor salivary glands do not develop hypertrophic, atrophic or hyperplastic changes. We detected a statistical difference between adult alcoholics without liver damage and adult alcoholics with liver damage. A diffuse mononuclear inflammatory cell infiltrate was distributed through the stroma or concentrated around the salivary ducts in both experimental and control samples. Acinar degeneration and hyperaemia were common changes observed in both experimental and control samples. Sialographic images from alcoholics demonstrated a similar morphology when compared with control patients, perhaps a reduction in secondary ducts diameter was observed in alcoholic patients. From this study we may conclude that minor salivary glands are scarcely affected by alcoholism, even when this disease has progressed to liver damage. PMID- 12893457 TI - Influence of age and of pre-treatment with D-cycloserine on the behavior of ethanol-treated rats tested in the elevated plus-maze apparatus. AB - There is evidence that ethanol is able to influence central functions through the antagonism of the NMDA-receptor system. It has been shown that this system is also involved in the modulation of anxiety-related behavior in rats. Recently, we observed gender- and age-related behavioral influences in rats tested on the elevated plus-maze apparatus The present study was undertaken in order to investigate: (1) the effects of ethanol (0.8, 1.0 or 1.2 g/kg, i.p.) on the behavior of male and female rats tested on the elevated plus-maze at 2, 3, 4 or 5 months of age; (2) the effect of the pre-treatment with D-cycloserine (3.0 or 6.0 mg/kg), an agonist of the glutamate NMDA-receptor system, 30 min before the ethanol (1.2 g/kg) injections, in rats tested in the elevated plus-maze at 2 months or 4 months of age. The results demonstrated that ethanol did not affect the time spent and the frequency of entries on the open arms of the elevated plus maze in rats tested at 2 months of age, but increased these parameters in older animals. Moreover, the results showed that D-cycloserine, at doses that did not affect the behavior of control animals, antagonized the increased frequency of entries and time spent on open arms produced by ethanol in rats tested at 4 months of age. Our results suggest an age-related influence on the anxiolytic action of ethanol in rats tested in the elevated plus-maze. Moreover, the results suggest that the NMDA-receptor system can be involved in this effect, and strengthens the evidence for the participation of the NMDA-receptor system in anxiety-related behavior. PMID- 12893458 TI - Role of parasympathetic (vagal) cardiac control in elevated heart rates of smokers. AB - Smokers may develop chronic increases in cardiac rate and alterations in cardiovascular control. If the increased mean heart rate (HR) in cigarette smokers is due in part to a deficit in vagal cardiac rate control, this should be reflected in a decreased amplitude of respiratory sinus arrhythmia (RSA). To test this hypothesis we studied 36 smokers and 36 non-smokers, matched for age, race, gender and blood pressure. All subjects were studied in the supine and seated positions. Mean heart rate was determined from the ECG during 30 s of quiet breathing; RSA was determined for 10 consecutive deep (>50% vital capacity) slow (5-7/min) breaths. Mean HRs in smokers were significantly higher than in non smokers, but the increases in mean HRs evoked by a shift from the supine to seated position were lower in smokers than in non-smokers, suggesting that chronic tobacco use may alter the relative contributions of sympathetic and parasympathetic control of cardiac rate. Because neither the RSAs nor the position-dependent increase in RSA were different between smokers and non smokers, we conclude that the elevated mean HRs in smokers were not the result of decreased respiratory or vasomotor modulation of vagal cardiac control, but instead were the result primarily of sympathetic stimulation. PMID- 12893459 TI - Acute ethanol-mediated insulin resistance in the rat: the role of ethanol oxidation. AB - Ethanol causes an acute and profound insulin resistance in humans and in the rat. Recent studies indicate that defects in skeletal muscle glucose uptake and utilization make a major contribution to this insulin resistance. In this study, we used the euglycaemic hyperinsulinaemic clamp to examine the role that hepatic ethanol oxidation via alcohol dehydrogenase (ADH) plays in the acute insulin resistance caused by ethanol in the rat. Treatment with the ADH inhibitor 4 methylpyrazole (4-MP) failed to abolish the insulin resistance as expressed as a decrease in the rate of glucose infusion required to maintain euglycaemia (GIR). A decrease in GIR was also observed in response to tert-butanol, an alcohol that is not a substrate for hepatic ADH. These results indicate that oxidation via ADH is not a prerequisite for the inhibition by ethanol of whole-body glucose utilization. In a separate study, we examined the relationship between blood ethanol concentration and GIR in order to determine the potency of ethanol in causing insulin resistance. These experiments showed that even at low blood concentrations (<2 mM), ethanol caused a profound decrease in GIR, similar in magnitude to that observed at higher blood concentrations (approximately 40 mM) PMID- 12893460 TI - Antibodies to morphine as indicators of chronic morphine intoxication and impaired immune reactivity. AB - Chronic morphine treatment of rats for 2 and 4 weeks led to an increase in morphine-binding cells in the spleen, despite the general reduction of mononuclear cell content in the spleen and thymus. Simultaneously, serum antibodies to morphine (AbM) in haemagglutination titres 1:20 and higher appeared in 50% and 80% of animals, respectively. Animals with high titres of AbM had much weaker immune response to a thymus-dependent antigen (sheep red blood cells). In humans chronic opiate intoxication was accompanied by significant reduction of functional activity of T-lymphocytes, which was especially dramatic in addicts with high levels of antibodies to morphine: high incidence of infection was obvious in the latter group. Results suggest that high levels of antibodies to morphine serve as indicators, not only of chronic morphine intoxication, but of impaired immune reactivity, especially involving T-cell functional activity. PMID- 12893462 TI - Quantitative trait locus gene mapping: a new method for locating alcohol response genes. AB - Alcoholism is a multigenic trait with important non-genetic determinants. Studies with genetic animal models of susceptibility to several of alcohol's effects suggest that several genes contributing modest effects on susceptibility (Quantitative Trait Loci, or QTLs) are important. A new technique of QTL gene mapping has allowed the identification of the location in mouse genome of several such QTLs. The method is described, and the locations of QTLs affecting the acute alcohol withdrawal reaction are described as an example of the method. Verification of these QTLs in ancillary studies is described and the strengths, limitations, and future directions to be pursued are discussed. QTL mapping is a promising method for identifying genes in rodents with the hope of directly extrapolating the results to the human genome. This review is based on a paper presented at the First International Congress of the Latin American Society for Biomedical Research on Alcoholism, Santiago, Chile, November 1994. PMID- 12893463 TI - Are tobacco and alcohol use related to Alzheimer's disease? A critical assessment of the evidence and its implications. AB - Elucidation of the association of tobacco and alcohol use with Alzheimer's disease (AD) may advance etiological hypotheses and provide a theoretical basis for treatment. There is currently no cure or effective treatment for AD, and no cause has been established. Pharmacological evidence supports a plausible biological mechanism for the involvement of tobacco use: nicotine compensates for some of the cholinergic deficits observed in AD. Epidemiological evidence, however, is inconsistent, although recent meta-analyses also support a protective effect. Although smoking per se is certainly not advocated, further investigation of a potential protective effect of nicotine on AD is warranted. Pharmacological studies implicate alcohol use as a possible risk factor for AD; the epidemiological studies are again inconclusive. Alcohol consumption is associated with daily smoking and smokers are, in turn, more likely to consume alcohol. Since tobacco use may decrease the risk of developing AD and alcohol use may increase it, it is important to consider these two substances together: the effect of one may negate the other. This literature review critically evaluates the evidence for an association of tobacco and alcohol use with AD and identifies key issues for further research. PMID- 12893464 TI - Ethanol dosage regimes in studies of ethanol toxicity: influence of nutrition and surgical interventions. AB - In this review we consider some of the practical facets of acute and chronic drug regimes. In particular, we illustrate our arguments with specific reference to alcohol and draw attention to methodological constraints that might alter biochemical or physiological processes. These includes the imposition of nutritional abnormalities and surgical procedures. These two areas are highlighted because there is evidence to show that they have marked influences on metabolic parameters, exemplified by tissue protein synthesis. In general, there is no controversy as to methods for acute studies with alcohol, although some reports have failed to investigate whether intravenous, intragastric or intraperitoneal regimes more accurately mimic the clinical situation. With regard to chronic feeding regimes, evidence is provided to support the argument that, currently, the most appropriate feeding protocol ensures that both control and treated groups receive identical amounts of nutrients albeit with differences in the calories apportioned to ethanol, which is substituted by isocaloric glucose or other carbohydrates in controls. However, there are still other methods being employed: these are subject to misinterpretation as the principle of ensuring that control and ethanol-fed rats receive identical amounts of nutrients, is ignored. In this review we also draw attention to the fact that surgical procedures, which are often employed to facilitate the measurement of body parameters (for example, implantation of cannulae), themselves alter tissue metabolism. The importance of this relates to the concept of metabolic superimposition, which implicates interacting responses when two or more stresses (i.e. surgery and drug administration) are superimposed. PMID- 12893465 TI - Brain regional pharmacology of GABA(A) receptors in alcohol-preferring AA and alcohol-avoiding ANA rats. AB - Compounds interacting with the GABA(A) receptor system modulate voluntary alcohol consumption in alcohol-preferring AA (Alko, Alcohol) rats. Therefore, we compared the central GABA(A) receptor pharmacology of the AA rats to that of their counterpart, alcohol-avoiding ANA (Alko, Non-Alcohol) rats with receptor autoradiography. Total flumazenil-sensitive [(3)H]Ro 15-4513 binding to the benzodiazepine site of GABA(A) receptor was slightly lower in the hippocampus, striate cortex and lateral hypothalamus of the AA than ANA rats. The proportions of zolpidem- and diazepam-sensitive components were similar in both rat lines. Basal picrotoxin-sensitive [(35)S]TBPS binding to the convulsant site of GABA(A) receptor was similar in most regions between the rat lines, but the up-modulation of the binding by 10 microM diazepam in the hippocampal, amygdaloid and entorhinal cortical areas was greater in the AA than ANA rats. These results do not reveal any general genetic defect in the GABA(A) receptors of AA or ANA rats, but the regional profile of the ligand binding differences between the lines, especially in the coupling of the benzodiazepine and chloride channel sites, suggests receptor subtype-specific changes in brain regions implicated in behavioural reward and anxiolysis. PMID- 12893466 TI - Performance in the cross-maze and slip funnel tests of four pairs of rat lines selectively bred for divergent alcohol drinking behavior. AB - Alcohol naive rats from lines genetically selected for high and low alcohol drinking and founded on either Wistar (P/NP lines) and distinct heterogeneous stocks (AA/ANA and replicate HAD/LAD lines) were tested in the explorative cross maze and the inescapable slip funnel. Rats of the low alcohol-consuming ANA, NP, LAD1 and LAD2 lines all exhibited a shorter latency before initiating exploration of the maze than did their high alcohol-consuming counterparts (66, 51, 33 and 51% of the values for AA, P, HAD1 and HAD2 lines, respectively). Significant line differences were also found with the slip funnel test (AA > ANA for time in a sprawling posture; P > NP but HAD1 < LAD1 and HAD2 < LAD2 for time escaping), but the directions of line differences were not consistently related to those in alcohol drinking. PMID- 12893467 TI - No association between D3 dopamine receptor (DRD3) alleles and cocaine dependence. AB - Cocaine is thought to act in the brain primarily by blocking dopamine re-uptake. The dopamine D3 receptor (genetic locus DRD3) is localized to brain regions that have been implicated in the reinforcing effects of a number of substances of abuse, including cocaine. The DRD3 coding region contains a polymorphism identifiable as a polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP). This polymorphism leads to an amino acid substitution at position 9 in the extracellular N-terminus of the D3 dopamine receptor. We examined alleles of the DRD3 gene in cocaine dependence using a genetic association strategy in samples of 62 white and 62 black cocaine-dependent individuals. Comparisons were made with local (Connecticut) control subjects for both groups, and with a larger sample of literature controls (for the white subjects) and a contrast group of schizophrenic patients (for the black subjects). No association was found between cocaine dependence and DRD3 alleles in either group (Bonferroni corrected). There was a significant difference in allele frequency between whites and blacks. These results are consistent with no role for genetic variation of the D3 dopamine receptor in susceptibility to cocaine dependence. PMID- 12893468 TI - Ethanol acutely impairs glycogen repletion in skeletal muscle following high intensity short duration exercise in the rat. AB - Ethanol is recognized to affect adversely carbohydrate metabolism in skeletal muscle. This paper seeks to establish whether ethanol acutely impairs glycogen repletion during recovery from high intensity short duration exercise in the rat. High intensity exercise caused the massive mobilization of glycogen stores in muscles rich in type IIa and IIb fibres and marked increases in plasma and muscle lactate levels. During the 30-minute recovery period, there was substantial glycogen repletion in these muscles in both the ethanol-treated and control rats. Ethanol, however, was associated with reduced glycogen resynthesis in both the tibialis anterior (by 22%) and red gastrocnemius (by 31%) muscles but not in the white gastrocnemius muscle. This reduction in post-exercise glycogen deposition was accompanied by decreased lactate disposal and elevated plasma glucose levels. These effects of ethanol on glycogen repletion may involve interactions with hepatic gluconeogenesis, glucose uptake and utilization in muscle, muscle glycogen synthesis and lactate glyconeogenesis. The ethanol-mediated impairment in post-exercise glycogen repletion may have important implications for the pathogenesis of chronic alcoholic skeletal myopathy. PMID- 12893469 TI - Cardiovascular risk factors in young drug addicts. AB - Since the type of life-style often encountered in drug addicts is purported to influence their risk of cardiovascular disease, it seemed of interest to carry out a descriptive study on risk factors in young addicts. Twenty-four young intravenous drug addicts, mean age 29 years and mean body mass index (BMI) 22.5 (kg/m(-2)), were compared with 24 healthy people matched for age, gender and BMI. Both groups responded to a questionnaire on life-style risk factors. In addition, fasting blood samples were investigated for several coronary risk factors. Drug addicts had smoked more cigarettes (17 vs. 3 cigarettes/day) for a longer time period (15 vs. 5 years) than control subjects. They also had higher serum activities of liver enzymes related to alcohol abuse than controls. Level of physical activity and indicators of dietary intake of fruit, vegetables and salt were not significantly different between the groups. Blood lipid values, total cholesterol (5.1 vs. 5.6 mmol/l), and HDL-cholesterol (1.2 mmol/l), apolipoproteins, and an atherogenic index reflecting the balance between the atherogenic low density lipoproteins (LDL) and the antiatherogenic fraction (HDL), as well as the smoking-related factors fibrinogen and TBARS, were all similar in both groups. The study indicates that the prevalence of many known cardiovascular risk factors was similar in drug addicts and matched controls. PMID- 12893470 TI - Interaction of mazindol with alcohol in mice. AB - Combined anorectic-alcohol misuse is a prevalent problem in Brazil. In order to understand better the interactive effects of ethanol (EtOH) and mazindol (MZ), we examined the effects of EtOH (1.2 g/kg) and MZ (5.0 mg/kg) given alone or in combination, on mouse behaviour. The results indicate that EtOH plus MZ induces a significantly greater increase in locomotor activity of mice than either constituent alone. However, no such interactive effect was detected in the place preference and in the plus-maze test of anxiety. MZ given alone was found to increase the locomotor activity and to possess rewarding effects as measured in the place preference conditioning. At the dose selected, EtOH alone showed anxiolytic and rewarding effects. These results provide some evidence of increased behavioural effects in mice due to combinations of EtOH and MZ. These findings suggest that a major determinant of combined anorectic-alcohol misuse may be the increased stimulating effects produced by such combination. PMID- 12893471 TI - Beta-carbolines in chronic alcoholics. PMID- 12893473 TI - Current state of knowledge about the mechanisms of alcohol tolerance. AB - Far from being a simple homeostatic response to the presence of ethanol in the brain, tolerance is now recognized as a complex process which can develop within various time frames (acute, rapid, chronic) and in which the drug interacts with various environmental and cognitive factors, including associative and operant learning. A major question is whether the acute form is an innate adaptive response, which is converted into the rapid and chronic forms by the action of these external influences. So far, all behavioral and neuropharmacological manipulations that alter chronic tolerance also modify rapid and acute tolerance in similar ways. These include lesions of serotonergic forebrain projections, blockade of NMDA-type glutamate receptors and calcium "L" channels, central or peripheral injection of vasopressin and blockade of vasopressin V1 receptors. Cytochemical and immunofluorescence studies, combined with use of retrograde tracers, indicate the existence of a septohippocampal circuit which may mediate the interactions of these diverse elements in the production and maintenance of tolerance. There is limited evidence that development of tolerance leads to increased consumption of ethanol in experimental animals, but the clinical significance of these findings remains to be proven. PMID- 12893474 TI - Metabolic effects of alcohol on skeletal muscle. AB - The purpose of this review is to summarize our current understanding of the acute and chronic interactions between alcohol and nutrient metabolism in skeletal muscle. Insulin is well known to play an important regulatory role in nutrient, especially glucose, uptake and utilization in skeletal muscle. Several studies have shown that alcohol can acutely reduce the normal metabolic responses of skeletal muscle to the action of insulin. The most obvious of these is an acute impairment in glucose metabolism associated with alcohol consumption. While the exact mechanism(s) underlying this acute insulin resistance is presently unclear, several possible factors are discussed in this review. In contrast to these short term effects, the effects of alcohol on skeletal muscle insulin sensitivity in chronic alcohol abusers are not as well established. Chronic alcohol abuse is known to be associated with skeletal myopathies, believed to result from alcohol induced abnormalities in muscle protein synthesis. Finally, the alcohol-mediated impairments of many aspects of skeletal muscle metabolism are discussed in relation to the insulin resistance associated broad spectrum of common lifestyle related disorders, including non-insulin dependent diabetes mellitus and obesity, the consequences of which may be important to the pathogenesis of alcohol-related diseases. PMID- 12893475 TI - Transfected cells for study of alcohol actions on GABA(A) receptors. AB - The ability to transfect and express foreign genes in cultured cells and Xenopus oocytes has contributed immensely to our knowledge of drug-receptor interactions and signal transduction following receptor activation. These techniques are now being used to study acute and chronic effects of ethanol on neurotransmitter receptors and cellular signaling. This article reviews the use of transfected cells to study ethanol actions on the GABAA receptor. PMID- 12893476 TI - Oral and dental health in the alcohol misuser. AB - This review explores the relationship between oral and dental health and disease in substance misusers. Although it focuses on oral cancer, we also examine the relationship of misuse with dental caries and periodontal diseases. It is difficult to separate out the inter-related aetiological factors such as the use of alcohol and tobacco (smoked and chewed). They probably act in a synergistic manner. PMID- 12893477 TI - The acute and chronic effects of alcohol upon cardiac nucleotide status. AB - The aim of the investigation was to ascertain the biochemical and morphological basis for the functional impairments in the heart due to alcohol. In chronic studies rats were fed a nutritionally complete liquid diet containing 35% of total calories as ethanol, controls were pair-fed identical amounts of the same diet in which ethanol was replaced by isocaloric glucose. In acute studies rats were injected with ethanol at a dose of 75 mmol/kg body weight. Pre-treatment of acute ethanol-dosed rats with cyanamide (ALDH inhibitor) was designed to raise acetaldehyde levels. In chronic studies ventricular adenine nucleotides, ATP, ADP and AMP; NAD(+), ATP ratios and the energy charge showed no alteration after 6 weeks of alcohol feeding. Light and electron microscopy sections indicated very little structural damage to muscle fibres and organelles (especially the mitochondria) in both atria and ventricles. Ventricular fibre diameters, throughout the different ranges, showed no significant differences between chronically alcohol and control-fed rats. In acute studies an increase in ventricular AMP levels (micromoles/g wet weight) occurred following cyanamide and cyanamide + ethanol treatment (+57%, p < 0.025 and +76%, p<0.01, respectively), but not as a consequence of ethanol alone. Cyanamide+ethanol caused marked elevation in ADP levels (+28%, p < 0.05) and again ethanol was without effect. ATP and GTP levels were not altered by any of the acute treatments. The energy charge was slightly reduced in both cyanamide and cyanamide+ethanol groups (-8%, p < 0.01 and -7%, p < 0.05, respectively), but not by ethanol alone. In conclusion, chronic alcohol appears to have minimal effects upon cardiac nucleotides which suggest that possible adaptive mechanisms are induced during the 6-week period and that alternative pathways other than defects in adenine nucleotide concentrations are involved in the pathogenesis of AHMD. The acute study suggests that the heart is resilient to toxic levels of alcohol and acetaldehyde in terms of ATP and GTP levels, despite the elevated AMP, ADP and GDP levels. PMID- 12893478 TI - Children with Fetal Alcohol Syndrome in North Dakota: a case control study utilizing birth certificate data. AB - A retrospective case control study utilizing birth certificate data in a population of children with Fetal Alcohol Syndrome (FAS) or Fetal Alcohol Effect (FAE) and controls in North Dakota was completed. Using the North Dakota FAS registry, 97 cases of FAS and FAE aged birth-18 years of age were identified. The North Dakota Department of Vital Records then searched for the child's birth certificates. For each case child, four controls were selected from the birth records. The controls were of the same race, sex, month and county of birth as the cases. Birth certificates for 68 children were identified, 44 with FAS and 24 with FAE. When compared with the FAE group, the FAS group had lower birth weights and mothers who began prenatal care later in pregnancy. The FAS/FAE group combined had mothers who were older, were more likely to be unmarried, had less weight gain during pregnancy, started prenatal care later in pregnancy and had fewer prenatal visits compared to controls. The FAS/FAE children had lower birth weights and higher rates of sibling deaths. The use of birth certificate data is a useful data source to compare maternal, paternal and prenatal characteristics for a population of children with FAS/FAE. PMID- 12893479 TI - Studies on the sensitivity to alcohol and alcohol dependence. AB - The role of genetic and environmental factors in the development of alcohol dependence is beginning to be investigated in Brazil. This paper reviews recent studies on genetic aspects of alcohol use and alcohol dependence, with emphasis on those carried out in Brazilian populations. Most findings replicated studies in developed countries, while some indicate new possible venues for investigation in the future. They underscore the need for a cross-cultural and interethnic approach to biomedical aspects of alcohol use and dependence. PMID- 12893480 TI - A study of hepatobiliary function in chronic heroin smokers. AB - The controversy over the relationship between heroin addiction and hepatocellular dysfunction was examined by assay of serum activities of aspartate and alanine aminotransferases, total protein, albumin and globulin in 48 chronic heroin smokers and 33 controls. Biliary secretion in the addicts was compared to controls by assay of serum bilirubin and its conjugated and unconjugated fractions. Serum aminotransferases, total protein, albumin and globulin did not differ significantly between the addict and the control groups (p < 0.05), whereas total bilirubin and its conjugated fraction were significantly higher in the addicts (p < 0.02). The results indicate that chronic abuse of heroin by smoking affects biliary secretion but exercises little influence over hepatocellular function. PMID- 12893483 TI - Alcohol misuse and renal damage. AB - Recent clinical and experimental studies have demonstrated that the habitual consumption of large amounts of ethanol has deleterious effects on the kidney. A variety of tubular defects have been described in patients with chronic alcoholism. Evidence is emerging that tubular dysfunction has an important pathophysiological role in a wide range of electrolyte and acid-base disturbances commonly observed in these patients, and possibly in alcohol-induced bone disease. These renal abnormalities are often reversible, disappearing with abstinence. However, since 1990 a few cases of a syndrome of acute tubular necrosis due to binge drinking of ethanol in the absence of other evident nephrotoxic mechanisms, or in association with the use of nonsteroidal anti inflammatory drugs, have been reported. A link between glomerulonephritis and alcoholism has become evident. IgA nephropathy has been demonstrated at autopsy in 64% of chronic alcoholics and, more recently, the association between alcoholism and postinfectious glomerulonephritis has been described. Structural and functional abnormalities of the kidney are reported with increasing frequency in the fetal alcohol syndrome seen in children who have been prenatally exposed to ethanol. In addition, over the last few years experimental studies in vitro or in animal models have provided information about the biochemical and molecular basis of alcohol-induced injury to kidney. It is hoped that future experimental and clinical research will provide us with a more comprehensive knowledge of the mechanisms of renal damage in alcohol misuse. PMID- 12893484 TI - The mu opioid receptor: from molecular cloning to functional studies. AB - Opioids have been used and abused by humans for centuries. The mu opioid receptor represents the high affinity binding site for opioid narcotics with high abuse liability such as morphine, codeine and fentanyl. Heroin (diacetylmorphine), a semi-synthetic derivative of morphine, crosses the blood-brain barrier more readily than morphine due to its increased hydrophobicity. Once in the brain heroin is hydrolyzed to morphine, which acts at the mu opioid receptor and results in euphoria, thus conferring the reinforcing properties of heroin. Using molecular biology techniques, the mu opioid receptors from several species have been cloned. This article reviews recent progress in this area, with respect to the two major cellular functions of the mu opioid receptor: reduction of intracellular cAMP concentration by inhibiting adenylyl cyclase activity, and inhibition of neuronal firing by modulating membrane ion channels. PMID- 12893485 TI - Cocaine and cardiovascular toxicity. AB - Over the past 10 years a great deal has been learned about the cardiovascular effects of cocaine. In particular, the acute effects of cocaine have been studied extensively. Upon acute administration cocaine increases blood pressure and heart rate, primarily through an action on the sympathetic nervous system. Cocaine also suppresses the baroreflex response and vagal tone, further contributing to its effects on heart rate. At the same time cocaine is increasing the work-load on the heart it induces coronary artery vasoconstriction, potentially leading to cardiac ischemia. At higher doses cocaine can depress ventricular function and slow electrical conduction in the heart. Both these effects appear to be mediated by cocaine's local anesthetic action. The effects of cocaine mediated by the sympathetic nervous system are greatly reduced in anesthetized animals. Further, when cocaine is administered repeatedly over a short period of time, acute tolerance can develop to the sympathomimetic effects of cocaine. In contrast, the effects of cocaine mediated by its local anesthetic action do not appear blunted by anesthesia or susceptible to acute tolerance. With chronic administration, higher doses appear to induce tolerance while lower doses may induce sensitization to cocaine's sympathomimetic effects. Cocaine also induces a variety of pathological changes in the heart, including myocardial contraction band necrosis and ventricular hypertrophy. These effects of cocaine on the heart can all contribute to potentially lethal cardiovascular events. In addition to the effects of cocaine alone, the metabolites of cocaine may also contribute to cocaine's cardiovascular toxicity, and both licit and illicit drugs used in combination with cocaine might potentially alter its cardiovascular effects. PMID- 12893486 TI - Functional coupling of a mu opioid receptor to G proteins and adenylyl cyclase: modulation by chronic morphine treatment. AB - A cloned mu opioid receptor was used to study its coupling to signal transduction pathways and its involvement in morphine-induced opioid dependence in stably transfected Chinese hamster ovary (CHO) cells. Membrane binding assays with a mu selective agonist [(3)H]DAMGO showed that one cell line expresses a high level of mu opioid receptors with a B(max) of approximately 630 fmol/mg membrane protein and a K(d) of 0.47 nM for DAMGO. Stimulation of the transfected cells with DAMGO led to an increase in the low K(m) GTPase activity, indicative of activation of guanine nucleotide regulatory proteins (G proteins), and this effect was blocked by the opioid antagonist naloxone. In addition, binding of the mu opioid receptor to DAMGO was affected by GTP and nonhydrolizable GTP analogs, Gp(NH)pp and GTP gamma-S. These results suggest a functional coupling between the mu opioid receptor and G proteins. Furthermore, DAMGO treatment of the cells produced a dose-dependent inhibition of the intracellular cyclic adenosine monophosphate (cAMP) level, with an EC(50) value of approximately 30 nM. Chronic treatment of the cells with morphine not only elevated the basal and forskolin-stimulated cAMP levels after morphine withdrawal, but also increased the extent of the DAMGO induced reduction of intracellular cAMP levels. The whole cell binding assay with [(3)H]DAMGO, on the other hand, did not detect receptor down-regulation after chronic morphine treatment. These results suggest that chronic morphine treatment may trigger a compensatory mechanism in cellular signaling pathways to offset the inhibitory input from the mu receptor without down-regulation of the surface receptor number, and that withdrawal of chronic inhibition leads to elevated activities of adenylyl cyclase to provide a basis for system sensitization. PMID- 12893487 TI - Induction of heat shock protein (HSP)-70 in posterior cingulate and retrosplenial cortex of rat brain by dizocilpine and phencyclidine: lack of protective effects of sigma receptor ligands. AB - The role of sigma receptors in the induction of heat shock protein (HSP)-70 by non-competitive N-methyl-Daspartate (NMDA) receptor antagonists (+)-MK-801 (dizocilpine) and phencyclidine (PCP) was studied. HSP-70 is induced in the posterior cingulate and retrosplenial cortex of rat brain 24 hours after a single administration of dizocilpine (1 mg/kg) or PCP (50 mg/kg). The induction of heat shock protein HSP-70 by dizocilpine or PCP was attenuated partially by pre treatment with the antipsychotic drug haloperidol (3 mg/kg, i.p., 15 minutes previously). However, pre-treatment with high potent and selective sigma receptor ligands, 4-phenyl-4-(1-phenylbutyl)piperidine (4-PPBP, 3 mg/kg, i.p., 15 minutes previously) and N,N-dipropyl-2-[4-methoxy-3-(2-phenylethoxy)phenyl]-ethylamine monohydrochloride) (NE-100, 3 mg/kg, i.p., 15 minutes previously) did not alter the induction of HSP-70 by dizocilpine or PCP. These findings suggest that sigma receptors may not play a significant role in the induction of HSP-70 by non competitive NMDA receptor antagonists dizocilpine and PCP, and that protective effects of haloperidol on induction of HSP-70 protein by dizocilpine or PCP may be due to other effect(s) except sigma receptors. PMID- 12893488 TI - Alcohol and glucose metabolism in skeletal muscles in the rat. AB - Ethanol is known to cause an acute and profound insulin resistance in man and the rat primarily via effects on glucose utilization. This paper examines the nature of these inhibitory effects on whole-body glucose utilization using the euglycaemic hyperinsulinaemic clamp in the conscious unrestrained rat. We confirm that ethanol infusion causes an acute insulin resistance, the rate of glucose infusion required to maintain euglycaemia (GIR) being decreased markedly by ethanol. To ensure that the GIR is a measure of whole-body glucose disposal, glucose turnover and hepatic glycogen levels were measured. These studies showed that ethanol totally suppressed hepatic glucose production. The reduction in GIR is associated with marked decreases in glucose uptake and glycogen synthesis in most skeletal muscles. In oxidative but not in non-oxidative muscles, the activation of glycogen synthase in response to insulin was decreased by ethanol, suggesting that a defect in glycogen synthase activation may be responsible for the decrease in glycogen synthesis. The basis of the inhibitory effects of ethanol on insulin-stimulated glucose metabolism in muscle is unknown, but may involve membrane-associated impairments in insulin signalling and/or the glucose transport system. PMID- 12893489 TI - Characterization of contractile and non-contractile protein synthesis in the stomach, small and large intestine and caecum of the rat, and response to acute ethanol dosage. AB - An investigation was made into the relative composition and synthesis rates of smooth muscle contractile proteins in vivo in different regions of the rat gastrointestinal tract. There was considerable homogeneity in the composition of sarcoplasmic proteins in the small bowel (i.e. 54-58 mg/g) but considerable variability in the large bowel, i.e. highest in the caecum (97 mg/g) and lowest in the colon (21 mg/g). The myofibrillary protein concentration was constant throughout the gastrointestinal tract, i.e. 20-34 mg/g. Stromal fractions varied from 6 to 39 mg/g and was highest in the cardiac region of the stomach and lowest in the duodenum. Fractional rates of protein synthesis were measured with a flooding dose of L-[4-(3)H]phenylalanine. In control rats synthesis rates in sarcoplasmic protein fractions were relatively much higher (43-107%/day) than myofibrillar (27-52%/day) or stromal (6-26%/day) proteins. Fractional rates of stromal protein synthesis showed the greatest variability while myofibrillar synthesis rates the least, throughout the alimentary tract. Jejunal seromuscular layer myofibrillar proteins had the highest synthesis rates (49%/day). In response to acute ethanol injection, protein synthesis in all jejunal fractions fell by 20-30%. Contractile and non-contractile proteins from the cardiac region of the stomach, duodenal seromuscular layer and large bowel seromuscular layer were insensitive to ethanol administration. Protein synthesis of sarcoplasmic proteins from the antrum, ileum seromuscular layer and myofibrillar proteins from the ileum seromuscular layer and caecum were also significantly depressed as a result of ethanol treatment. PMID- 12893490 TI - Beta-carbolines in chronic alcoholics following trauma. AB - In our society every second polytraumatized patient is a chronic alcoholic. A patient's alcohol-related history is often unavailable and laboratory markers are not sensitive or specific enough to detect alcohol-dependent patients who are at risk of developing alcohol withdrawal syndrome (AWS) during their post-traumatic intensive care unit (ICU) stay. Previously, it has been found that plasma levels of norharman are elevated in chronic alcoholics. We investigated whether beta carbolines, i.e. harman and norharman levels, could identify chronic alcoholics following trauma and whether possible changes during ICU stay could serve as a predictor of deterioration of clinical status. Sixty polytraumatized patients were transferred to the ICU following admission to the emergency room and subsequent surgery. Chronic alcoholics were included only if they met the DSM-III R and ICD-10 criteria for alcohol dependence or chronic alcohol abuse/harmful use and their daily ethanol intake was > or =60 g. Harman and norharman levels were assayed on admission and on days 2, 4, 7 and 14 in the ICU. Harman and norharman levels were determined by high pressure liquid chromatography. Elevated norharman levels were found in chronic alcoholics (n = 35) on admission to the hospital and remained significantly elevated during their ICU stay. The area under the curves (AUC) showed that norharman was comparable to carbohydrate-deficient transferrin (CDT) and superior to conventional laboratory markers in detecting chronic alcoholics. Seventeen chronic alcoholics developed AWS; 16 of these patients experienced hallucinations or delirium. Norharman levels were significantly increased on days 2 and 4 in the ICU in patients who developed AWS compared with those who did not. An increase in norharman levels preceded hallucinations or delirium with a median period of approximately 3 days. The findings that elevated norharman levels are found in chronic alcoholics, that the AUC was in the range of CDT on admission and that norharman levels remained elevated during the ICU stay, support the view that norharman is a specific marker for alcoholism in traumatized patients. Since norharman levels increased prior to the onset of hallucinations and delirium it seems reasonable to investigate further the potential role of norharman as a possible substance which triggers AWS. PMID- 12893491 TI - Pharmacokinetic and pharmacodynamic interactions in an outpatient maintenance therapy of intravenous heroin users with levomethadone. AB - The plasma levels of 42 patients on a levomethadone maintenance treatment programme for intravenous heroin users were measured before and, respectively, 1, 2 or 4 hours after oral routine administration and related to the individual additional drug usage (detected by urine drug screening), liver function, side effects and withdrawal symptoms. In general, accelerated levomethadone metabolism induced by additional misuse of benzodiazepines, barbiturates and opiates resulted in significantly lower plasma levels of the substitute. In particular, high gamma-glutamyltransferase activity was related to benzodiazepine consumption. On the other hand, an impaired liver function reflected by increased beta-globulins resulted in an insufficient body clearance and drug accumulation. Major side effects, such as sweating, were not related to plasma levels whereas withdrawal symptoms like diarrhoea or "feeling cold" correlate with lower plasma concentrations. It is concluded that polydrug misuse in the methadone maintenance therapy creates a vicious circle of enzyme induction, thus increasing "instrumental drug utilization". However, underestimated maintenance dosage may lead to additional drug consumption resulting, finally, in therapeutic failure. PMID- 12893493 TI - Lessons from Thomas Alva Edison --the greatest American inventor-- to surgical investigators. PMID- 12893494 TI - John Syng Dorsey --writer of the first American systematic surgical textbook. PMID- 12893495 TI - Composite tissue allotransplantation. AB - Composite tissue allotransplantation (CTA) recently took its first steps in the clinical arena in 1998 with the successful hand transplant performed in Lyons, France. That single operation represented a culmination of many years of laboratory research in multiple fields involving integumentary/musculoskeletal transplantation. Here we review the prerequisite developments in the field of immunology, microsurgery, and pharmacotherapy that helped bring CTA to clinical reality. This new field still has many unanswered questions which are addressed below. Additionally, new evolving research in CTA is also discussed. PMID- 12893496 TI - Adenoviral-mediated gene transfer of interleukin-2 in a human breast cancer cell line and a fresh primary breast cancer culture. AB - Adenovirally-mediated cytokine gene transfer has proven safe in the treatment of metastatic breast cancer. Unfortunately, the optimal conditions for gene transfer in the human breast remain largely unknown. Viral-mediated gene transfer was studied in a human breast cancer cell line and a fresh primary breast cancer culture using a type five adenoviral vector (AD5) containing the human interleukin 2 (IL-2) gene driven by a cytomegalovirus (CMV) promoter (AD5.CMV IL2). IL-2 production was measured using an enzyme-linked immunosorbent assay (ELISA). In the human breast cancer cell line (MCF-7), IL-2 production increased logarithmically with viral dose and demonstrated peak production at 2000 ng/10(6) cells/24 h using a multiplicity of infection (MOI) of 3000:1. Transduction at a higher MOI resulted in cell death. IL-2 concentration reached over 2000 ng/ml 2 days after transduction and peaked 13 days after transduction at 5700 ng/ml. IL-2 levels declined thereafter. A fresh primary breast cancer culture, transduced with Ad5.CMV-IL2 at an MOI of 1000:1, secreted IL-2 at 15 ng/24 h 1 day after transduction and peaked at 85 ng/24 h 5 days after transduction. Adenoviral mediated gene transfer was accomplished in breast cancer cells with high efficiency across a wide range of conditions. The optimal IL-2 dose required to maximally stimulate the immune system remains unknown. PMID- 12893497 TI - Usefulness of bovine pericardium as interpositional graft in the surgical repair of nasal septal perforations (experimental study). AB - A 2.5-cm nasal septal perforation was performed in 18 pigs and repaired as follows: group I (n = 6), septal perforation without treatment; group II (n = 6), surgical repair with interpositional graft of glutaraldehyde-preserved bovine pericardium (GPBP); group III (n = 6), surgical repair with interpositional graft of lyophilized GPBP (LGPBP). The animals were evaluated clinically and radiologically (x-ray and CT scan) 2 days before surgery, daily during the first postoperative week, and weekly during the next 6 months. At the end of the study the animals were euthanized with an overdose of pentobarbital. Macroscopic and microscopic examination of the grafts and nasal septum was performed. All the animals survived the surgical procedure. Five pigs in group I showed persistence of the septal perforation. All the animals in groups II and III showed total closure of the septal perforation, with the presence of fibrotic tissue on the pericardial grafts as well as in the septal cartilage, and overall good healing. In conclusion, GPBP and LGPBP are adequate materials that can be used as interpositional grafts in the surgical closure of septal perforations in pigs PMID- 12893498 TI - Metabolic alterations of skeletal muscle tissue after prolonged acute ischemia and reperfusion. AB - Acute tissue ischemia is usually followed by considerable disturbances of cellular metabolism that often lead to cell death. Reperfusion improves cellular function by withdrawing the toxic products of ischemia and providing energy sources, although sometimes it worsens it. The purpose of this experimental work is to study the metabolic disturbances in skeletal muscle tissue of canines after prolonged acute ischemia in relation to the values of certain substances (ATP, lactate, pyruvate, the ratio of lactate to pyruvate [L/P], and glucose) and whether the alterations in the values of these substances could be used as prognostic indices of the magnitude of tissue damage and the possibility of inverting it. We used 15 mongrel dogs. Complete acute ischemia was induced in the right lower limb lasting 12 h. Reperfusion also lasted for 12 h. The left lower limb was used as reference value. Before the beginning of ischemia, at (1/2), 1, 6, and 12 h after the induction of ischemia, and at (1/2), 1, 6, and 12 h after the restoration of circulation, blood samples, and tissue biopsies were obtained from the healthy and the experimental limb for the measurement of ATP, lactate, pyruvate, the ratio of L/P, and glucose. From the statistical analysis of the values of the controlled parameters the following were concluded: The changes in ATP, lactate, pyruvate, and the L/P ratio in the venous blood of the experimental limb and in the intracellular space of the suffering skeletal muscle could be used as indices to evaluate ischemic injury to the skeletal muscles, the course of its development, and the possibility of reversal after reperfusion. PMID- 12893499 TI - The role of the spleen on colonic anastomotic healing. AB - The role of the spleen on wound healing remains unclear. This study investigates the effect of splenectomy on the healing of colonic anastomoses. Twenty-six Wistar rats were assigned into four groups: sham, splenectomy, anastomoses, and splenectomy and anastomoses. The rats underwent a standardized left colonic resection and primary anastomoses, and/or splenectomy. Bursting pressure and hydroxyproline content were used to evaluate anastomotic healing, five days postoperatively. No differences were found in the bursting pressure and hydroxyproline content between the groups. The present results indicate that splenectomy has no negative effect on the healing of colonic anastomoses in rats. PMID- 12893500 TI - Plasma D-lactate levels in diagnosis of appendicitis. AB - We investigated the possible use of D-lactate as a predictor in the diagnosis of appendicitis. C-reactive protein level (CRP) and leukocyte counts were also evaluated. Venous blood D-lactate, CRP, and leukocyte counts were measured preoperatively in 53 patients undergoing surgery for appendicitis, as well as in 20 healthy subjects. Levels of all three parameters in the surgical patients were significantly higher than in the control group (p <.05). Previous studies have shown that venous D-lactate is more specific to the intestine than CPR or leukocyte count. Based on our data, venous D-lactate, which had the lowest false negative rate among these laboratory parameters, may be a useful diagnostic marker for appendicitis. None of these parameters were helpful in identifying the type of the appendicitis. PMID- 12893502 TI - Heterogeneity of obsessive-compulsive disorder: a literature review. AB - Significant advances have been made in characterizing the phenomenology and psychobiology of obsessive-compulsive disorder (OCD) in recent years. In many ways, such advances suggest a conceptualization of OCD as a relatively homogeneous neuropsychiatric entity, underpinned by particular mechanisms that manifest in universal symptoms. Nevertheless, some data have pointed to the heterogeneity of this disorder. A computerized literature search (MEDLINE: 1964 2001) was used to collect studies addressing the heterogeneity of OCD. In addition, reviews of the phenomenology, psychobiology, family studies, and treatment of OCD were examined in an attempt to collate data addressing this issue. There is a growing consensus that some subtypes of OCD are valid and provide a useful means of integrating data on its symptomatology, neurobiology, and treatment response; for example, OCD with comorbid tics is characterized by earlier onset, a particular range of OCD symptoms, and worse response to selective serotonin reuptake inhibitors. The heterogeneity of OCD has important clinical and research implications. PMID- 12893503 TI - Efficacy of combined therapy and pharmacotherapy for depressed patients with or without personality disorders. AB - In general, depressed patients with personality pathology--Axis II disorders- respond less well or less quickly to the various kinds of individual treatment that are available, whether pharmacotherapy, psychotherapy, or both combined. This article sets forth the results of a six-month, randomized clinical trial of antidepressants and combined therapy in ambulatory patients with major depression and a baseline score of at least 14 on the 17-item Hamilton Rating Scale for Depression (HAM-D-17). The presence or absence of Axis II pathology was determined on the basis of a self-report version of the International Personality Disorder Examination. The study's antidepressant protocol provided for three successive steps in case of intolerance or inefficacy: fluoxetine, amitriptyline, and moclobemide. In addition to pharmacotherapy, the combined-therapy condition included 16 sessions of Short Psychodynamic Supportive Psychotherapy. Efficacy of the therapy provided was assessed using the HAM-D-17 and also other instruments. According to the results in secondary analyses, it emerged that combined therapy was more effective than pharmacotherapy for depressed patients with personality disorders. Combined therapy was not more effective than pharmacotherapy alone for depressed patients without personality disorders. It is recommended that depressed patients with comorbid personality pathology should be treated with combined therapy, with the focus of psychotherapy being not on the patient's symptoms and complaints, but on all aspects of the patient's actual relationships. PMID- 12893504 TI - "My Face Is My Fate": biological and psychosocial approaches to the treatment of a woman with obsessions and delusions. PMID- 12893505 TI - Introduction: common mental disorders, primary care, and the global mental health research agenda. PMID- 12893506 TI - Primary care services for depression in Harare, Zimbabwe. PMID- 12893507 TI - Common mental disorders and primary health care: current practice in low-income countries. PMID- 12893508 TI - Fertilization rates of small-head sperm in conventional IVF and ICSI. AB - Five men produced 23 ejaculates in 23 assisted reproductive technology cycles for semen analysis. In 11 of the 14 ICSI cycles and in 5 of the 9 IVF cycles, small head sperm were found in more than 70% of the cells, having a length of < 3.5 micro m and a width of < 2 micro m. In 6 of the 14 ICSI cycles, the embryologist who performed the ICSI was not alerted to e presence of small-head sperm. Subsequent fertilization rate was significantly lower than a) the fertilization rate of the remaining 5 ICSI cycles that acted as a control, in which the embryologist was alerted to the presence of small-head sperm, and b) was lower than the 3 ICSI cycles with normal sperm head (p < .05). The 8 cell embryo and blastocyst rates in the former group were also decreased, although not significantly, compared to the latter groups. Likewise, in the 5 IVF cycles with small-head sperm, the fertilization rate was significantly lower than in the 4 IVF with normal sperm head (p < .05). Embryologists should be alerted when a high percentage of small-head sperm are detected during routine semen analysis because they may be associated with reduced fertilization and embryo outcome. PMID- 12893509 TI - Serum-free culture of rhesus monkey embryonic stem cells. AB - Previous studies have shown that the maintenance and proliferation of undifferentiated rhesus monkey embryonic stem (rES) cells requires medium supplemented with fetal bovine serum (FBS). Due to the uncharacterized composition and variation in serum nature, the present study aimed to replace the serum-containing medium with a serum-free medium in the rES cell culture. The results showed that after the initial 48-h culture in the routinely used serum containing medium, rES cells can grow and proliferate for a prolonged period in the serum-free medium composed of DMEM supplemented with a cocktail of BSA, IGF 1, TGF-alpha, bFGF, aFGF, estradiol, and progesterone. rES cells cultured in the serum-free medium maintained high level of alkaline phosphatase activity and OCT4 level. There was no indication of differentiation as judged by the marker gene expression of all three embryonic germ layers and trophoblast. In addition, serum free culture would not affect the passage capacity and differentiation potential of rES cells. This work will facilitate the future study of induced differentiation of rES cells and other applications. PMID- 12893510 TI - Asthenozoospermia: analysis of a large population. AB - Spermatozoa travel a long distance to meet and fertilize the oocyte, so sperm motility is a requisite for normal fertilization. Asthenozoospermia, or low sperm motility, is a common cause of human male infertility. This is a retrospective study (1992-1999) to document the prevalence of this pathology in infertile men and to clarify the probable factors associated to its etiology. The prevalence was 18.71% for asthenozoospermia and 63.13% for asthenozoospermia associated with oligo- and/or teratozoo-spermia; thus, 81.84% of the studied samples had altered motility. Leukocytospermia, the ratio of germ cells/sperm, anti-sperm antibodies, consistency, biochemical markers of accessory sex glands, and sperm response after swim-up were studied in normospermic (N), asthenozoospermic (A), and combined asthenozoospermic (C) samples. No significant difference was found in the frequency of leukocytospermia among groups. The rate of germ cells/(spermatozoa + germ cells) between C and N (p < .01) and C and A (p < .01) was statistically different, while no difference was found on comparing N and A. MAR-test over 40% was found in 6% of the A samples and 7.6% of the C, while no positive values were observed in the N group. The percentage of hyperviscous samples was higher in the low sperm motility samples than in the normal group. Data on concentration of the biochemical markers seem to be decreased in asthenozoospermia. Pure and combined asthenozoo-spermia showed different behavior in sperm recovery after swim-up. Two different asthenozoospermias could be defined: the pure one where sperm environment is involved (immunological factor, hyperviscosity, and secretory gland function) and the combined, where the testis is comprised. PMID- 12893511 TI - Prognostic value of evaluation of total number of morphological anomalies in 100 sperm in semen of infertile men. AB - In this study, the evaluation of the total number of morphological anomalies per 100 sperm was more reliable, as far as potential fertility is concerned than the concentration of sperm per milliliter, the percentage of motile sperm, and the percentage of sperm with normal or abnormal morphology. This parameter allows for discrimination between subfertile and fertile semen and therefore can help in the selection of men who require assisted reproduction techniques. PMID- 12893512 TI - Antioxidant activity of seminal plasma in fertile and infertile men. AB - This study was conducted to evaluate and compare the total antioxidant capacity among fertile and infertile men. Thirty infertile patients and 20 fertility proven healthy donors with normal sperm analysis were included in the study. Total antioxidant capacity, zinc and fructose levels of seminal plasma, and various sperm parameters were compared among fertile controls and idiopathic infertility patients prospectively. The mean antioxidant capacity of fertile controls (2.02 +/- 0.16 mmol/L) was significantly higher than that of the infertile patients group (1.78 +/- 0.23 mmol/L) (p < .01). Furthermore, asthenozoospermic and asthenoteratozoospermic groups had significantly lower mean antioxidant values (1.73 +/- 0.11 and 1.64 +/- 0.13, respectively) when compared to fertile control group (p < .01). The mean fructose level was significantly lower in the fertile control group and mean zinc level was significantly lower in the entire infertile group. On the other hand, antioxidant capacity is positively correlated to sperm motility (p = .001). Decreased antioxidant capacity was associated with impaired sperm function as a result of either increased ROS production or insufficient antioxidant capacity. PMID- 12893513 TI - Intractable bladder tamponade due to left internal pudendal and obturator arterial bleeding induced by erection: a superselective arteriography followed by transcatheter arterial embolization. AB - Bladder tamponade is an uncommon clinical symptom among men suffering during sexual intercourse. The authors report on a 46-year-old man for whom this symptom appeared 4-5 years prior to hospitalization. Angiography confirmed the presence of left pudendal and obturator arterial bleeding, and embolotherapy of the internal pudendal and obturator arteries was performed. There was no mortality, or limb loss or sexual potency loss at follow-up. PMID- 12893514 TI - Blood zinc and copper concentrations in criminal and noncriminal schizophrenic men. AB - This study investigated the effects of zinc and copper on the behavior of schizophrenic patients by comparing blood zinc and copper levels in criminal and noncriminal schizophrenic patients. Of the total 88 subjects, 44 were patients with schizophrenia and no criminal record who were being treated at the Elazig Mental Hospital and 44 were schizophrenic patients who had committed a crime and were sent to the same hospital upon a court order for monitoring, detention, and treatment. Of the subjects with criminal records, 31 (70%) had committed a violent crime and 13 (30%) had committed a nonviolent crime. The mean plasma zinc value was 68 +/- 1.55 microg/dL in the criminal subjects and 81 +/- 2.73 microg/dL in the noncriminal subjects ( p = .001). The mean serum copper value was 104 +/- 1.80 microg/dL in criminal subjects and 93 +/- 2.92 microg/dL in noncriminal subjects (p = .02). Mean plasma zinc values were significantly lower in criminal subjects when compared to noncriminal subjects, while mean serum copper values were significantly higher in criminal subjects than noncriminal subjects. PMID- 12893515 TI - Temporal trends in human semen parameters in New England in the United States, 1989-2000. AB - The current study explores trends in semen parameters in New England in the United States. A retrospective review was performed of 551 semen analysis records from 1989 to 2000 at Vincent Memorial Andrology Laboratory of Massachusetts General Hospital. After age adjustment, semen pH and motility significantly increased 0.05 units/year and 2.33%/year, respectively, while sperm with normal morphology decreased 0.33%/year. Sperm concentration showed a small upward trend. The year of birth in the present study ranged from 1932 to 1981; 2% were born between 1932 and 1941, 13% between 1942 and 1951, 48% between 1952 and 1961, 36% between 1962 and 1971, and 1% were born between 1972 and 1981. There were significant positive relationships between year of birth and semen volume (0.04 mL/1-year interval increase in year of birth) and motility (0.61 percent/1-year interval increase in year of birth), as well as with sperm concentration and morphology. Overall, there were temporal and year of birth trends in several human semen parameters. PMID- 12893516 TI - Thyroid hormones: their role in testicular steroidogenesis. AB - Thyroid hormones are important for growth and development of many tissues. Altered thyroid hormone status causes testicular abnormalities. For instance, juvenile hypothyroidism/neonatal transient hypothyroidism induces macroorchidism, increases testicular cell number (Sertoli, Leydig, and germ cells) and daily sperm production. Triiodothyronine (T3) receptors have been identified in sperm, developing germ cells, Sertoli, Leydig, and peritubular cells. T3 stimulates Sertoli cell lactate secretion as well as mRNA expression of inhibin-alpha, androgen receptor, IGF-I, and IGFBP-4. It also inhibits Sertoli cell mRNA expression of Mullerian inhibiting substance (MIS), aromatase, estradiol receptor, and androgen binding protein (ABP) and ABP secretion. T3 directly increases Leydig cell LH receptor numbers and mRNA levels of steroidogenic enzymes and steroidogenic acute regulatory protein. It stimulates basal and LH induced secretion of progesterone, testosterone, and estradiol by Leydig cells. Steroidogenic factor-1 acts as a mediator for T3-induced Leydig cell steroidogenesis. Although the role of T3 on sperm, germ, and peritubular cells has not yet been completely studied, it is clear that T3 directly regulates Sertoli and Leydig cell functions. Further studies are required to elucidate the direct effect of T3 on sperm, germ, and peritubular cells. PMID- 12893517 TI - Fertility after chemotherapy for testicular cancer. AB - Disseminated testicular cancer has largely become curable with cisplatin-based chemotherapy. The prospect of fertility after treatment is an important consideration for both patients and clinicians. While there may be an irreversible impairment of spermatogenesis at a cumulative cisplatin dose of greater than 400 mg/m2, a low sperm count does not necessarily appear to prevent fatherhood. This review summarizes currently available data on the effects of chemotherapy on male fertility and steps that can be taken to preserve fertility in this patient population. PMID- 12893518 TI - Cryosurgery for prostate cancer. AB - Choice of management for patients with prostate cancer is influenced by patient and disease characteristics and life expectancy. Management options include expectance (watchful waiting), radical prostatectomy, external beam radiotherapy, brachytherapy, and cryosurgical ablation of the prostate (CSAP). The role of cryotherapy in the management of prostate cancer is still evolving. Continued research has allowed the introduction of efficient and safe cryosurgical equipment exemplified by the current third-generation cryosurgical machines. CSAP can be performed in an ambulatory surgery setting or as inpatient surgery with overnight stay. The procedure is performed under continuous ultrasonic monitoring. Mature data from the use of second-generation cryosurgical equipment indicate that CSAP is an effective therapeutic modality for managing patients with prostate cancer. Current data with the third-generation cryosurgical equipment are not mature. However, the favorable side effect profile and the good early responses seem to indicate that this modality will have a prominent role in the management of patients with prostate cancer. PMID- 12893519 TI - Use of cultured precision-cut rat lung slices to study the in vitro induction of pulmonary cytochrome P450 forms. AB - 1. The aim was to investigate the effects of some cytochrome P450 (CYP) enzyme inducers on CYP1A and CYP2B subfamily forms in cultured precision-cut rat lung slices. 2. Precision-cut lung slices were prepared from male Sprague-Dawley rats and cultured for 24 and/or 48 h in medium containing 0-20 micro g ml(-1) Aroclor 1254 (ARO), 0-50 micro M beta-naphthoflavone (BNF) and 0-50 micro M benzo(a)pyrene (BP). 3. Treatment with ARO, BNF and BP produced significant increases in lung slice whole homogenate 7-ethoxyresorufin O-deethylase activity. 4. Levels of CYP1A1 apoprotein were markedly increased in lung slice microsomes after treatment for 48 h with either 10 micro g ml(-1) ARO or 5 micro M BNF. In contrast, neither ARO nor BNF had any marked effect on levels of CYP2B1/2 apoprotein in 48-h cultured rat lung slice microsomes. 5. Real-time quantitative reverse transcription-polymerase chain reaction methodology (TaqMan) was used to quantify lung slice CYP1A1 and CYP2B1/2 mRNA levels. Rat lung slice CYP1A1 mRNA levels were increased up to 8.3-fold after treatment for 24 h with 2 and 10 micro g ml(-1) ARO, 0.5 and 5 micro M BNF, and 20 micro M BP. In contrast, treatment with 10 micro g ml(-1) ARO produced only a small 1.6-fold increase in CYP2B1/2 mRNA levels. 6. Precision-cut lung slices are a useful model in vitro system for the assessment of the effects of chemicals on pulmonary CYP forms. PMID- 12893520 TI - Relative receptor expression is a determinant in xenobiotic-mediated CYP3A induction in rat and human cells. AB - 1. Species differences in xenobiotic-mediated transcriptional activation of CYP3A genes are known to exist. These differences are proposed to be due, in part, to host cell differences. 2. Host cell effects were investigated by trans-species transient transfection of reporter genes containing either the rat CYP3A23 or human CYP3A4 proximal promoters into human HepG2 and rat FaO and H4IIEC3 hepatoma cells. HepG2 and FaO cells supported activation of both CYP3A constructs by xenobiotics in a species-specific manner, whereas H4IIEC3 cells were non permissive. 3. The mRNA complement of the cell lines was then quantified by semiquantitative RT-PCR for adult CYP3As (CYP3A23, CYP3A4/5), steroid hormone receptors (constitutive androstane receptor, glucocorticoid receptor-alpha, pregnane X receptor) and transcription factors (Hepatic nuclear factor 4alpha, retinoid X receptor). 4. Principal component analysis of absolute receptor levels demonstrated a wide scattering, with no coherent pattern. In contrast, PCA of relative receptor ratios segregated H4IIEC3 cells from all other samples. 5. The observation is confirmed that species differences in response to xenobiotics are a result of host cell environment. In addition, new evidence is provided to support the hypothesis that in addition to individual receptor activation profiles, the relative abundance of steroid hormone receptors that control CYP3A gene expression play an important role in this observed species difference. PMID- 12893521 TI - In vitro activation of 7-benzyloxyresorufin O-debenzylation and nifedipine oxidation in human liver microsomes. AB - 1. The effects of substrate concentration and enzyme source (human liver microsomes and recombinant cytochrome P450s, CYP) on the activation of 7 benzyloxyresorufin O-debenzylation and nifedipine oxidation were investigated. 2. 7-Benzyloxyresorufin O-debenzylase activity in human liver microsomes was inhibited by a monoclonal antibody against CYP2B6 and a polyclonal antibody against CYP3A2 by 53-69 and 19-44%, respectively, suggesting that CYP2B6 and CYP3A4 mainly catalyse 7-benzyloxyresorufin O-debenzylation in human liver microsomes. 3. 7-Benzyloxyresorufin O-debenzylase activity at 0.2-5 micro M substrate concentrations in human liver microsomes was increased by the addition of alpha-naphthoflavone, quinidine, testosterone and progesterone, and the V(max) of 7-benzyloxyresorufin O-debenzylation increased with increasing alpha naphthoflavone concentrations, whereas the K(m) remained constant. Additionally, 7-benzyloxyresorufin O-debenzylation by recombinant CYP3A4 was increased by the addition of alpha-naphthoflavone, testosterone and progesterone but not by quinidine, whereas no chemicals tested could activate the O-debenzylation of 7 benzyloxyresorufin by CYP2B6. 4. The K(m) for nifedipine oxidation activity by CYP3A4 decreased by the addition of progesterone, whereas the V(max) remained constant. Quinidine and testosterone increased 7-benzyloxyresorufin O-debenzylase and nifedipine oxidase activities, respectively, in human liver microsomes, whereas activation was not observed in CYP3A4. 5. The results suggest that in vitro activation patterns are substrate dependent and that selection of the enzyme source can influence the activation phenomenon. PMID- 12893522 TI - Dexamethasone decreases plasma levels of the prochiral fenbendazole and its chiral and achiral metabolites in sheep. AB - 1. The effect of co-administration of either short- or long-acting formulations of DXM on hepatic function and the plasma pharmacokinetic behaviour of prochiral fenbendazole (FBZ) and its metabolites was evaluated in sheep. 2. Neither DXM treatment markedly affected any of the biochemical markers of hepatic function tested. In contrast, both formulations significantly modified the plasma pharmacokinetic behaviour of FBZ and its metabolites. 3. Plasma FBZ concentrations and the associated area under the time-concentration curves were significantly lower, although the plasma detection period was longer (72 versus 48 h) in the DXM pretreated animals compared with those given FBZ alone. 4. DXM also appeared to alter the pattern of FBZ absorption, possibly through effects on abomasal pH. The shape of the plasma concentration-time curves for oxfendazole (OFZ) and fenbendazole sulphone (FBZSO(2)) were similar to FBZ, raising the possibility that DXM treatment may have altered the liver biotransformation of the parent drug. 5. The concentrations of the (+) chiral metabolite of OFZ were significantly lower in DXM pretreated animals compared with those given FBZ alone. The trend was similar for the (-) antipode, although the differences between DXM pretreated and non-pretreated animals were not statistically significant. PMID- 12893523 TI - Bioavailability of dietary doses of 3H-labelled tea antioxidants (+)-catechin and (-)-epicatechin in rat. AB - 1. The bioavailability and pharmacokinetic characteristics of the tea antioxidants (+)-catechin and (-)-epicatechin were investigated in the rat following intake of dietary doses. 2. To achieve this objective, tritiated derivatives (tritium was incorporated at the 3-position of the heterocyclic ring) of these compounds were administered to rats orally and intravenously at dose levels equivalent to human dietary levels of intake. 3. Following intravenous administration of both compounds, about one-third of the dose was excreted in the urine and two-thirds in the faeces, indicating extensive biliary excretion. When the same doses were administered orally, only about 5% of the dose of each compound was recovered in the urine. 4. Comparison of the areas under the curve following oral and intravenous administration revealed that the bioavailability of both compounds was less than 5%. 5. Exchange of tritium with water in the blood occurred 3 h after oral, but not after intravenous, administration of the flavanols to rat. This is believed to represent microbial degradation of the compounds by the gut flora. 6. It was established that the bioavailability of the tea antioxidants (+)-catechin and (-)-epicatechin in the rat following intake of dietary doses was poor. PMID- 12893524 TI - Absorption, distribution, metabolism and excretion of telmesteine, a mucolitic agent, in rat. AB - 1. The metabolism and disposition of telmesteine, a muco-active agent, have been investigated following single oral or intravenous administration of (14)C telmesteine in the Sprague-Dawley rat. 2. (14)C-telmesteine was rapidly absorbed after oral dosing (20 and 50 mg kg(-1)) with an oral bioavailability of >90% both in male and female rats. The C(max) and area under the curve of the radioactivity in plasma increased proportionally to the administered dose and those values in female rats were 30% higher than in male rats. 3. Telmesteine was distributed over all organs except for brain and the tissue/plasma ratio of the radioactivity 30 min after dosing was relatively low with a range of 0.1-0.8 except for excretory organs. 4. Excretion of the radioactivity was 86% of the dose in the urine and 0.6% in the faeces up to 7 days after oral administration. Biliary excretion of the radioactivity in bile duct-cannulated rats was about 3% for the first 24 h. The unchanged compound mainly accounted for the radioactivity in the urine and plasma. 5. Telmesteine was hardly metabolized in microsomal incubations. A glucuronide conjugate was detected in the urine and bile, but the amount of glucuronide was less than 6% of excreted radioactivity. PMID- 12893525 TI - Species and gender differences in the formation of an active metabolite of a substituted 2,4-thiazolidinedione insulin sensitizer. AB - 1. The metabolism of a substituted 2,4-thiazolidinedione (P1) with dual PPARalpha/gamma activity was evaluated in male and female rats, dogs and monkeys. A para-hydroxylated metabolite (M1) with potent PPARgamma-selective agonist, was a major circulating drug-related component in female rats, dogs and monkeys, but not in male rats (M1-to-P1 exposure ratio of <1, 3-5, 5 and 5-11 in male rat, monkey, female rat, and dog, respectively). 2. M1 (%) formed in vitro (5, 53, 57 65, 67 and 67% in male rat, monkey, female rat, dog, and human liver microsomes, respectively), rank ordered with M1 (%) formed in vivo (24-45, 53-57, 78, 75-85%, for male rat, monkey, female rat and dog, respectively, after oral administration of P1). 3. The plasma clearance of M1 was higher in male rats (32 ml min(-1) kg( 1) compared with 6, 7 and 2 ml min(-1) kg(-1) in female rat, male monkey and male dogs, respectively). 4. The low amounts of M1 observed in male rats, with the appearance of products of the cleavage of the propyl group between the phenyl groups was probably due to the presence of the sex-specific CYP2C11, which cleaves P1 at the propyl bridge. None of the CYPs present in female rats cleaved P1 at this site and M1 was only produced by CYP2C6. In humans, only CYP2C8 and the polymorphic CYP2C19 produced M1. PMID- 12893526 TI - Metabolic fate of pitavastatin, a new inhibitor of HMG-CoA reductase: similarities and difference in the metabolism of pitavastatin in monkeys and humans. AB - 1. To elucidate any potential species differences, the in vitro metabolism of pitavastatin and its lactone was studied with hepatic and renal microsomes from rats, dogs, rabbits, monkeys and humans. 2. With the addition of UDP-glucuronic acid to hepatic microsomes, pitavastatin lactone was identified as the main metabolite in several animals, including humans. 3. Metabolic clearances of pitavastatin and its lactone in monkey hepatic microsome were much greater than in humans. 4. M4, a metabolite of pitavastatin with a 3-dehydroxy structure, was converted to its lactone form in monkey hepatic microsomes in the presence of UDP glucuronic acid as well as to pitavastatin. These results implied that lactonization is a common pathway for drugs such as 5-hydroxy pentanoic acid derivatives. 5. The acid forms were metabolized to their lactone forms because of their structural characteristics. 6. UDP-glucuronosyltransferase is the key enzyme responsible for the lactonization of pitavastatin, and overall metabolism is different compared with humans owing to the extensive oxidative metabolism of pitavastatin and its lactone in monkey. PMID- 12893527 TI - Substrate recognition by the mammalian proton-dependent amino acid transporter PAT1. AB - The PAT family of proton-dependent amino acid transporters has recently been identified at the molecular level. This paper describes the structural requirements in substrates for their interaction with the cloned murine intestinal proton/amino acid cotransporter (PAT1). By using the Xenopus laevis oocytes as an expression system and by combining the two-electron voltage clamp technique with radiotracer flux studies, it was demonstrated that the aliphatic side chain of L-alpha-amino acids substrates can consist maximally of only one CH2-unit for high affinity interaction with PAT1. With respect to the maximal separation between the amino and carboxyl groups, only two CH2-units, as in gamma aminobutyric acid (GABA), are tolerated. PAT1 displays no or even a reversed stereoselectivity, tolerating serine and cystein only in the form of D enantiomers. A methyl-substitution of the carboxyl group (e.g. O-methyl-glycine) markedly diminishes substrate affinity and transport rates, whereas methyl substitutions at the amino group (e.g. sarcosine or betaine) have only minor effects on substrate interaction with the transporter binding site. Furthermore, it has been shown (by kinetic analyses of radiolabelled betaine influx and inhibition studies) that the endogenous PAT system of human Caco-2 cells has very similar transport characteristics to mouse PAT1. In summary, one has defined the structural requirements and limitations thet determine the substrate specificity of PAT1. A critical recognition criterion of PAT1 is the backbone charge separation distance and the side chain size, whereas substitutions on the amino group are well tolerated. PMID- 12893528 TI - ER-to-Golgi transport: COP I and COP II function (Review). AB - COP I and COP II coat proteins direct protein and membrane trafficking in between early compartments of the secretory pathway in eukaryotic cells. These coat proteins perform the dual, essential tasks of selecting appropriate cargo proteins and deforming the lipid bilayer of appropriate donor membranes into buds and vesicles. COP II proteins are required for selective export of newly synthesized proteins from the endoplasmic reticulum (ER). COP I proteins mediate a retrograde transport pathway that selectively recycles proteins from the cis Golgi complex to the ER. Additionally, COP I coat proteins have complex functions in intra-Golgi trafficking and in maintaining the normal structure of the mammalian interphase Golgi complex. PMID- 12893529 TI - Regulated exocytosis and SNARE function (Review). AB - The pairing of cognate v- and t-SNAREs between two opposing lipid bilayers drives spontaneous membrane fusion and confers specificity to intracellular membrane trafficking. These fusion events are regulated by a cascade of protein-protein interactions that locally control SNARE activity and complex assembly, determining when and where fusion occurs with high efficiency in vivo. This basic regulation occurs at all transport steps and is mediated by conserved protein families such as Rab proteins and their effectors and Sec1/unc18 proteins. Regulated exocytosis employs auxiliary components that couple the signal (which triggers exocytosis) to the fusion machinery. At the neuronal synapse, munc13 as well as munc18 control SNARE complex assembly. Synaptotagmin and complexin ensure fast synchronous calcium-evoked neurotransmitter release. PMID- 12893530 TI - Cisternal maturation and vesicle transport: join the band wagon! (Review). AB - 'No cellular organelle has been the subject of as many, as long-lasting or as diverse polemics as the Golgi apparatus'. This statement was made by Whaley almost 30 years ago in the book The Golgi Apparatus and still holds true today, perhaps more then ever. Why is this? How come something as mundane as a series of intracellular membrane bound structures continues to fascinate and captivate a large section of the cell biology community? One simple reason (putting polemics aside) is that the secretory pathway appears deceptively simple. Once probed, however, it has a persistent habit of developing into an enigma. Is one then not closer than 30 years ago? In a sense yes, in that one has more components and a better understanding of inherent membrane dynamics, but it is still not known how newly synthesized proteins and lipids make their way from the ER to the plasma membrane. Is it by vesicles, cisternal carriers or transient tubular connections? It has also been learned that newly synthesized proteins are segregated away from the resident components throughout the pathway, but not how. Do coat proteins hold the key? It is understood that the cytoskeleton is important, but not really why. It is known that each Golgi stack is a fully functional unit, but not why stacks are connected laterally into a large ribbon (the Golgi apparatus). This review focuses on how proteins make their way through the pathway, a basic question that remains to be answered. PMID- 12893531 TI - Lipid microdomains, lipid translocation and the organization of intracellular membrane transport (Review). AB - Eukaryotic cells contain hundreds of different lipid species that are not uniformly distributed among their membranes. For example, sphingolipids and sterols form gradients along the secretory pathway with the highest levels in the plasma membrane and the lowest in the endoplasmic reticulum. Moreover, lipids in late secretory organelles display asymmetric transbilayer arrangements with the aminophospholipids concentrated in the cytoplasmic leaflet. This lipid heterogeneity can be viewed as a manifestation of the fact that cells exploit the structural diversity of lipids in organizing intracellular membrane transport. Lipid immiscibility and the generation of phase-separated lipid domains provide a molecular basis for sorting membrane proteins into specific vesicular pathways. At the same time, energy-driven aminophospholipid transporters participate in membrane deformation during vesicle biogenesis. This review will focus on how selective membrane transport relies on a dynamic interplay between membrane lipids and proteins. PMID- 12893532 TI - Modulation of entry of enveloped viruses by cholesterol and sphingolipids (Review). AB - Enveloped animal viruses infect host cells by fusion of viral and target membranes. This crucial fusion event occurs either with the plasma membrane of the host cells at the physiological pH or with the endosomal membranes at low pH and is triggered by specific glycoproteins in the virus envelope. Both lipids and proteins play critical and co-operative roles in the fusion process. Interactions of viral proteins with their receptors direct which membranes fuse and viral fusion proteins then drive the process. These fusion proteins operate on lipid assemblies, whose physical and mechanical properties are equally important to the proper functioning of the process. Lipids contribute to the viral fusion process by virtue of their distinct chemical structure, composition and/or their preferred partitioning into specific microdomains in the plasma membrane called 'rafts'. An involvement of lipid rafts in viral entry and membrane fusion has been examined recently. However, the mechanism(s) by which lipids as dynamic raft components control viral envelope-glycoprotein-triggered fusion is not clear. This paper will review literature findings on the contribution of the two raft associated lipids, cholesterol and sphingolipids in viral entry. PMID- 12893533 TI - Modulation of MRP1-like efflux activity in human erythrocytes caused by membrane perturbing agents. AB - The effect of membrane perturbing agents on the efflux (37 degrees C, 60 min) of the fluorescent probe 2', 7'-bis-(carboxypropyl)-5(6)-carboxyfluorescein (BCPCF) from human erythrocytes was studied. Several anionic amphiphiles (detergents) markedly inhibited BCPCF efflux (IC50 < or = 40 microM). Most zwitter-ionic amphiphiles were inefficient inhibitors. Non-ionic and cationic amphiphiles had minor effects or increased efflux. Of the aliphatic inhibitors, C12-homologues were the most efficient. Hexanol, ethanol, methyl-beta-cyclodextrin (MbetaCD) and diamide (+ washing) did not influence BCPCF efflux. It is suggested that amphiphiles affect BCPCF efflux by modulating multi-drug resistance protein 1 (MRP1, ABCC1) activity. A negative charge of amphiphiles is essential for the inhibitory effect, while alkyl chain length modulates inhibition. MRP1-mediated BCPCF efflux appears to be relatively insensitive to non-specific plasma membrane modification. PMID- 12893535 TI - Scanning mutagenesis studies of the M1 muscarinic acetylcholine receptor. AB - Following the solution of the structure of bovine rhodopsin by X-ray crystallography, it has been possible to build an improved homology model of the M(1) muscarinic acetylcholine receptor. This has been used to interpret the outcome of an extensive series of scanning and point mutagenesis studies on the transmembrane domain of the receptor. Potential intramolecular interactions enhancing the stability of the protein fold have been identified. The residues contributing to the binding site for the antagonist, N-methylscopolamine, and the agonist, acetylcholine have been mapped. The positively charged headgroups of these ligands appear to bind in a charge-stabilized aromatic cage formed by amino acid side chains in transmembrane (TM) helices 3, 6, and 7, while residues in TM 4 may participate in a peripheral docking site. Closure of the cage around the headgroup of acetylcholine may help to transduce binding energy into receptor activation, possibly disrupting a set of Van der Waals interactions between a set of residues underlying the binding site which help to constrain the receptor to the inactive state, in the absence of agonist. This may trigger the reorganization of a hydrogen bonding network between highly conserved residues in the core of the receptor, whose integrity is crucial for activation. PMID- 12893536 TI - Structure/activity relationships of M2 muscarinic allosteric modulators. AB - Allosteric modulation of G protein-coupled receptors has been intensively studied at muscarinic acetylcholine receptors. Findings made with archetypal allosteric agents such as gallamine, alcuronium, and bis(ammonio)alkane-type agents revealed that binding of orthosteric ligands that attach to the acetylcholine site can be allosterically decreased or increased or left unaltered in a subtype-selective fashion. Analyses of structure/activity relationships (SARs) help to elucidate the molecular events underlying the allosteric action and they may pilot the development of new allosteric agents with improved properties and therapeutic perspectives. With a focus on SARs, this review illustrates the principles of muscarinic allosteric interactions, gives an overview of SARs in congeners of archetypal allosteric agents, and considers the topology of M(2) muscarinic allosteric interactions that are characterized by divergent binding modes. PMID- 12893537 TI - Cellular signaling mechanisms for muscarinic acetylcholine receptors. AB - Signaling pathways for muscarinic acetylcholine receptors (mAChRs) include several enzymes and ion channels. Recent studies have revealed the importance of various isoforms of both alpha and betagamma subunits of G proteins in initiation of signaling as well as the role of the small monomeric G protein, Rho, in the activation of phospholipase D. Modulation of adenylyl cyclase activity by mAChRs appears more diverse as the interaction of various receptor subtypes with the many isoforms of the enzyme are studied. Both alpha and beta subunits of G(i/o) may be involved. Some mAChR responses arise through release of nitric oxide from nitrergic nerves, including salivary gland secretion and hippocampal slow wave activity. mAChRs utilize a variety of intracellular pathways to activate various mitogen-activated protein kinases. The kinases are involved in cholinergic regulation of kidney epithelial function, catabolism of amyloid precursor protein, hippocampal long-term potentiation, activation of phospholipase A(2), and gene induction. mAChR activation can also stimulate or inhibit cellular growth and apoptosis, dependent on prior levels of cellular activity. Modulation of ion channels by mAChR agonists appears increasingly complex, based on recent studies. K(+) channels may be activated by M(2) and M(4) mAChR stimulation, although in the rat superior cervical ganglion topographical constraints appear to limit the effect to the M(2) mAChR. Another ganglionic K(+) current, the M current, is inhibited by M(1) mAChR activation, but in murine hippocampus inhibition involves another receptor subtype. R-type Ca(2+) channels are both facilitated and inhibited by M(1) and M(2) mAChRs; facilitation being more pronounced with activation of M(1) mAChRs and inhibition with M(2) mAChRs. PMID- 12893538 TI - Pharmacological analysis of the contractile role of M2 and M3 muscarinic receptors in smooth muscle. AB - Muscarinic receptors expressed on smooth muscle cells are primarily of the M(2) and M(3) subtypes. The M(3) subtype triggers contraction through an interaction with G(q) proteins to stimulate phosphoinositide hydrolysis and mobilize Ca(2+). In contrast, activation of M(2) receptors modulates contraction by preventing relaxation or by potentiating M(3) receptor-mediated contractions, which enhances heterologous desensitization. These effects can be explained by the coupling of M(2) receptors to G(i) proteins that mediate an inhibition of adenylyl cyclase and calcium-activated potassium channels. The pharmacological antagonism of a response mediated through an interaction between M(2) and M(3) receptors has been shown to resemble the profile of the directly acting receptor (M(3)), primarily, and not that of the conditional receptor (M(2)). Evidence for a contractile role of the M(2) receptor has been obtained by inactivating its signaling pathway with pertussis toxin or by measuring contractile effects of muscarinic agonists after M(3) receptors have been covalently inactivated. Under these conditions, M(2) receptors have been shown to mediate an inhibition of the relaxant effects of agents, like isoproterenol, on the contractile effects of nonmuscarinic spasmogens. Muscarinic M(2) and M(3) receptor knockout mice are useful tools for exploring interactions between these receptors in smooth muscle. PMID- 12893539 TI - M1-M5 muscarinic receptor knockout mice as novel tools to study the physiological roles of the muscarinic cholinergic system. AB - A large body of evidence indicates that muscarinic acetylcholine receptors (mAChRs) play critical roles in regulating the activity of many important functions of the central and peripheral nervous systems. However, identification of the physiological and pathophysiological roles of the individual mAChR subtypes (M(1)-M(5)) has proven a difficult task, primarily due to the lack of ligands endowed with a high degree of receptor subtype selectivity and the fact that most tissues and organs express multiple mAChRs. To circumvent these difficulties, we used gene targeting technology to generate mutant mouse lines containing inactivating mutations of the M(1)-M(5) mAChR genes. The different mAChR mutant mice and the corresponding wild-type control animals were subjected to a battery of physiological, pharmacological, behavioral, biochemical, and neurochemical tests. The M(1)-M(5) mAChR mutant mice were viable and reproduced normally. However, each mutant line displayed specific functional deficits, suggesting that each mAChR subtype mediates distinct physiological functions. These results should offer new perspectives for the rational development of novel muscarinic drugs. PMID- 12893540 TI - Reconfiguring old age: elderly women and concerns over care in southeastern Botswana. AB - This paper explores recent changes in the negotiation of female old age in Southeastern Botswana. Over the past one-quarter century epidemiological and demographic transitions, increasing pressures on lay nursing care, changes in work, and the introduction of pensions have led to a fracturing of old age. The constituent elements of old age--senescence and social elderhood--have shifted at different tempos and in different directions in historically contingent ways. Using Margaret Lock's notion of "local biology," this paper explores transformations in the bio-social dialectic that have reconfigured and redefined the aging process. It argues that the "normal" physiology and social position of seniors have changed so that chronic illness is increasingly seen as part of "normal" old age, as is the lack of socio-economic and cultural power to command care. PMID- 12893541 TI - "Nobody gives a damn if I live or die": violence, drugs, and street-level prostitution in inner-city Hartford, Connecticut. AB - Drawing on the tenets of critical medical anthropology, this article illustrates the relation between violence, drug use, prostitution, and HIV risk in a group of 35 impoverished women living in inner-city Hartford, Connecticut. The study presented here provides an illustration of the role prostitution plays in the SAVA (Substance Abuse, Violence, and AIDS) syndemic as conceptualized by Singer (1996). By focusing on the life experiences of women engaged in street-level prostitution, this article attempts to fill the gaps in research that deals simultaneously with these mutually reinforcing epidemics. It shows that street walkers' continuous exposure to violence, both as victims and as witnesses, often leaves them suffering from major emotional trauma. In the absence of adequate support services, women who have been victimized may turn to drug use in an attempt to deal with the harsh realities of their daily lives. In turn, the need for drugs, coupled with a lack of educational and employment opportunities, may lead women into prostitution. Life on the street increases women's risk for physical, emotional, and sexual abuse as well as their risk for HIV/AIDS. Exposure to traumatic experiences deepens the dependence on drugs, completing a vicious cycle of violence, substance abuse, and AIDS risk. PMID- 12893542 TI - The acupuncture wars: the professionalizing of American acupuncture--a view from Massachusetts. AB - Since the 1970s acupuncturists in the United States have confronted the dilemma of how to define themselves not only as practitioners in relation to an evolving Americanized version of Chinese medicine but also with respect to definitions of biomedical professional identity, which are currently in flux. The central issue is that of professionalization. This study traces the process of professionalization through the initial reception of the modality; the first steps toward specialized training; and the further steps through professional associations, credentialing, and licensing. This process takes place within the broader social frame of fluctuating definitions of biomedical professionalism. It is within this context that acupuncturists are assessing role definition, status, and compensation. Part of the process also involves the renewed use of the clinical trial and the potential co-opting of acupuncture. The potential for resistance is tied in with alliances with holistic physicians and with acupuncturists' own defense of pluralism. PMID- 12893543 TI - Standards and medical education. PMID- 12893544 TI - BEME Guide no 3: systematic searching for evidence in medical education--Part 1: Sources of information. AB - Searching for evidence to inform best practice in medical education is a complex undertaking. With very few information sources dedicated to medical education itself, one is forced to consult a wide range of often enormous sources--and these are dedicated to either medicine or education, making a medical education search all the more challenging. This guide provides a comprehensive overview of relevant information sources and methods (including bibliographic databases, grey literature, hand searching and the Internet) and describes when they should be consulted. The process of constructing a search is explained: identifying and combining core concepts, using Boolean algebra and search syntax, limiting results sets, and making best use of databases' controlled vocabularies. This process is illustrated with images from search screens and is followed by numerous examples designed to reinforce skills and concepts covered. The guide has been developed from the ongoing experience gained from the systematic searches conducted for the Best Evidence Medical Education Collaboration, and concludes by looking ahead to initiatives that will shape future searching for medical education evidence. PMID- 12893545 TI - Twelve tips for developing a clinical teaching programme in a day surgery unit. AB - Healthcare policy in the UK is moving towards an expansion in day care services. As it becomes increasingly difficult to deliver clinical teaching to undergraduates in traditional inpatient venues, opportunities must be sought in ambulatory care. The proposed increased activity of day surgery units provides one such resource for the development of a structured clinical teaching programme. This paper highlights 12 tips for the preparation, delivery and evaluation of a clinical teaching programme in the day surgery unit. It describes the implications for staff and resources and indicates the educational opportunities that can be provided. PMID- 12893546 TI - Developing the teaching instinct. 8: Presentations. PMID- 12893547 TI - Managing and supporting medical education with a virtual learning environment: the Edinburgh Electronic Medical Curriculum. AB - Virtual learning environments (VLEs) can be a compelling and powerful way to support and manage contemporary medical education. A VLE purposively aligned to a course can integrate and normalize procedures and provide a central access point and reference mechanism for all of a course's component communities. The Edinburgh Electronic Medical Curriculum (EEMeC), developed in-house to support the Edinburgh under-graduate course, has proved to be a great success with students, teaching and administrative staff. It exists in a 'blended' relationship with the course, which uses both face-to-face and online delivery modes. This paper maps out the process of the EEMeC system's development and describes a number of factors that have contributed to its success. PMID- 12893548 TI - An Internet-based pediatric clerkship examination. AB - A traditional paper final exam was expensive and time consuming to create, administer and grade at two separate training sites. Our goal was to determine whether restructuring the final pediatric clerkship examination using Internet technology could lower costs and be more efficient, reliable and preferred by students. Costs and coordinator time were compared with the prior year of traditional testing. Student satisfaction was determined by an electronic post test survey. To evaluate for reliability, the scores on our Internet test were compared with United States Medical Licensing Examination (USMLE) shelf scores. An estimated 1200 US dollars per year of department costs was saved, and coordinator time was greatly reduced. The post-test survey indicated the students were very satisfied with this testing. When compared with USMLE scores the Cronbach's alpha was 0. 70, and the Pearson Correlation was r = 0.414 (p<0. 001). It is concluded that a well-designed Internet-based test has advantages over a traditional paper exam. It reduces incremental department costs, is reliable,preferred by students and can be given simultaneously at multiple locations. PMID- 12893549 TI - Representative survey of dental students about the present state of education, PC equipment and configuration requirements for a net-based dental education system. AB - A representative survey of 130 dental students revealed that the necessary conditions for using net-based education systems seem to exist, as 99% of all students have access to a PC, of which most (78%) own one. As most of them use the PC at least several times a week, they seem to be adequately skilled to deal with hard- and software. Internet and emailing are the most commonly used features of the PC after text documentation, therefore it can be inferred that almost all computers have Internet access. The students' requirements for the planned net-based dental education system are case documentation, multimedia documented work-flows, checklists related to workflows, a training tool for prosthetic therapy planning, practical advice and films about dental treatment methods, timetables and discussion fora. The net-based dental education system can support private study but may also be used for seminars in small groups. PMID- 12893550 TI - A case study of methods used to tackle a common pedagogic problem in medical and dental education: time pressure. AB - This case study describes how the common problems of time shortage, factual overload and curriculum overcrowding were tackled in one context, using good practice guidelines. It examines the impact of two hours of intensive, interactive teaching on the knowledge and attitudes of senior medical undergraduates at one United Kingdom school, and the effectiveness of two different ways of presenting information. The session raised student awareness of the importance to medical practice of the topic. There were statistically significant increases in the ability and knowledge of students over the period, as self-rated by students and assessed by the teacher; results of the impact on learning outcomes of different versions of a handout were not statistically significant. Further research on modes of presentation of information and their impact on forming and accessing illness scripts is suggested. Minority subjects in potentially overcrowded, time-pressured curricula can be effectively introduced in short time slots. PMID- 12893551 TI - Peer tutoring in patient-centred interviewing skills: experience of a project for first-year students. AB - Peer tutoring is a potentially valuable resource in higher education. There are few published accounts of the impact of peer tutoring in medical education. College-wide experience of peer tutoring together with difficulties recruiting medical teachers for a communication programme led to the development of a peer tutoring project. This paper reports the impact on first-year medical students of peer tutoring in patient-centred interviewing. After attending a preparatory workshop, third-year medical students co-facilitated their first-year colleagues in a session:Interviewing a Simulated Patient. First-year students completed written evaluations immediately after the session and two months later randomly selected students were assessed in patient-centred interviews. The impact of the peer-tutoring experience was evaluated by comparing these outcomes for students in groups co-facilitated by peer tutors with those who worked in groups facilitated by medical teachers. The eight learning objectives were completely met by more than 56% of students. However, there were statistically significant differences for four objectives with more students in groups facilitated by medical teachers completely meeting these objectives. Although the seven educational techniques used in the session were rated favourably by all students, two were rated as more effective in achieving the learning objectives by students in groups facilitated by medical teachers. Free-text comments revealed no differences between groups. Two months after the session, there were no differences between students in terms of interviewing skills as rated by trained observers and simulated patients, whilst simulated patients were more satisfied with interviews from students facilitated by peer tutors (p <0.02). Peer tutors can support the acquisition of basic patient-centred interviewing skills in first year medical students when contributing to one session of a structured programme. First-year students were receptive and willing to learn from their senior colleagues. PMID- 12893552 TI - The Supplemental Instruction Project: peer-devised and delivered tutorials. AB - The purpose of this study was to determine whether student devised and delivered supplemental instruction is beneficial and acceptable to first-year medical students. A student-run Supplemental Instruction Project (SIP) was developed and delivered by second-year medical students and offered free of charge to all first year medical students at Memorial University of Newfoundland taking the Integrated Study of Disease I course in 1999 and again in 2000. Small-group tutorials focused on subject material that second-year medical students identified as 'difficult'. Five 60- to 90-minute sessions covering topics in cardiology, nephrology and respirology were offered. Student and tutor perceptions about the project were collected using anonymous questionnaires. Students were quizzed before and after each tutorial session. Post-tutorial quiz scores were significantly greater than pre-tutorial scores. Student and tutor perceptions of SIP were positive. It is concluded that the SIP is an acceptable, practical and effective method to supplement delivery of challenging material to first-year medical students. PMID- 12893553 TI - An interprofessional problem-based learning course on rehabilitation issues in HIV. AB - This study examined students' perceptions of their learning through participation in an interprofessional problem-based course on rehabilitation and HIV. Students representing five health professions participated in an eight-week tutorial course. Qualitative analysis of journals that the students completed throughout the course, and of interviews of the students at completion of the course, revealed that they valued their learning experience. Students gained an appreciation of the roles of others and developed a sense of confidence through justifying their professional role. Through the interprofessional discussions, students were able to increase the breadth and depth of their learning and also gained a rehabilitation perspective. Learning related to HIV and rehabilitation is ideally suited to an interprofessional, problem-based environment. PMID- 12893554 TI - Measurement practices: methods for developing content-valid student examinations. AB - Measurement experts generally agree that a systematic approach to test construction will probably result in an instrument with sound psychometric properties. One fundamental method is called the blueprint approach to test construction. A test blueprint is a tool used in the process for generating content-valid exams by linking the subject matter delivered during instruction and the items appearing on the test. Unfortunately, this procedure as well as other educational measurement practices is often overlooked A survey of curriculum administrators at 144 United States and international medical schools was conducted to assess the importance and prevalence of test blueprinting in their school. Although most found test blueprinting to be very important, few require the practice. The purpose of this paper is to review the fundamental principals associated with achieving a high level of content validity when developing tests for students. The short-term efforts necessary to develop and integrate measurement theory into practice will lead to long-term gains for students, faculty and academic institutions. PMID- 12893555 TI - Medical curricula in China and the USA: a comparative study. AB - To examine the differences between the content of curriculum of medical schools in China and the USA the authors compared the curricula of 82 Chinese and 125 American medical colleges. Great disparities were found in the framework of medical curricula, processes of teaching learning, types and numbers of preclinical courses, course hours, laboratory and practical training hours, and key disciplines of clinical practice. A significant difference was also found in the number of courses, lectures and laboratory and clerkship hours among Chinese medical colleges. The authors conclude that medical colleges in China should look again at the type of courses offered, revise curricular frameworks and reform the teaching learning process and approaches. New courses should be introduced, new content should be added to existing courses, and appropriate importance should be attached to clinical practice of subjects such as family and community medicine. To improve the quality of medical education, Chinese medical colleges need national medical education standards compatible with international standards. PMID- 12893556 TI - Teaching in Spanish medical schools. AB - The study reported here assessed the current situation of the teaching of medicine, the healthcare facilities available and the teaching staff employed at Spanish medical schools, on the basis of a national survey devised by the Conferencia Nacional de Decanos de Facultades de Medicina. There was a 100% response rate from the 27 medical schools. Twenty-three of these schools introduced new study plans between 1993 and 2000. In all cases these plans are basically discipline oriented, with different levels of integration between the subjects offered. The study also analysed teaching in bioethics, healthcare management, experience in primary care and alternative or complementary medicine. In the 2000-01 academic year there were 4471 first-year students; as for the healthcare facilities available, there were 67,331 hospital beds in 112 hospitals and 234 Primary Care Centres. There was an average of 4.49 teaching beds per student. There were 3081 full and associate professors, roughly half of whom were teaching in the preclinical cycle and roughly half in the clinical cycle, and 4492 clinical assistant professors. The average student-professor ratio at Spanish medical schools is currently 3.02: 1.0, with variations of between 6.22 and 1.04. PMID- 12893557 TI - An operational measure of physician lifelong learning: its development, components and preliminary psychometric data. AB - Despite the emphasis placed on physicians' lifelong learning, no psychometrically sound instrument has been developed to provide an operational measure of the concept and its components among physicians. The authors designed this study to develop a tool for measuring physician lifelong learning, to identify its underlying components and to assess its psychometric properties. A 37-item questionnaire was developed, based on a review of literature and the results of two pilot studies. Psychometric analyses of the responses of 160 physicians identified 19 items that were included in the Jefferson Scale of Physician Lifelong Learning. Factor analysis of the 19 items showed five meaningful factors that were consistent with the definition and major features of lifelong learning. They were 'need recognition', 'research endeavor', 'self-initiation', 'technical skills' and 'personal motivation'. The method of contrasted groups provided evidence in support of the validity of the five factors. The factors' reliability was assessed by coefficient alpha. It is concluded that lifelong learning is a multifaceted concept, and its operational measure is feasible for evaluating different educational programs and for studying group differences among physicians. PMID- 12893558 TI - The use of the opinion leader in continuing medical education. AB - This paper describes a process evaluation of an opinion leader (OL) project for a geriatric medical education program in dementia. Structured interviews with OLs were conducted to understand their experience as formal OLs from their initial training to the project's completion. Thirteen of 15 physicians involved in an OL project were interviewed. The adoption of a formal educational OL role was not an easy transition for most identified OLs. Some physicians struggled with the role and would have preferred more specific guidance. For this subject some content expertise was felt to be essential. Strong project leadership and some measures of success are important to sustain the OL commitment to a project. More attention needs to be devoted to the appropriate selection and training of OLs for educational projects to ensure that they have the relevant clinical expertise and skills to be effective and feel comfortable adopting a formal OL role. PMID- 12893559 TI - Scholarly development of clinician faculty using evidence-based medicine as an organizing theme. AB - While scholarship is a critical component of the mission of all academic departments, little is known about how to approach the scholarly development of clinician faculty. The authors developed a thematically based faculty development curriculum with two primary components: evidence-based clinical practice and critical appraisal of current clinical research. They took a task management approach to reducing anticipated barriers to program success by instituting a regularly scheduled seminar series, providing evidence-based reading materials, recruiting clinician faculty as speakers for the clinical presentations, and providing continuing medical education credits for seminar attendance. This case study illustrates a faculty development curriculum that stimulated scholarly discussion and research activities in the authors' clinician faculty. PMID- 12893564 TI - The design and construction of an interactive website concerning biomedical photography. AB - The purpose of this communication is to make readers aware of what the authors believe is an important online resource about medical and scientific photography for doctors, scientists and students. It is a website freely accessible and its URL is http://msp.rmit.edu.au. The site is designed as a resource base: it is not meant to be a 'course' but the reader will find much practical information about technique and applications of scientific imaging methods. The site is currently a comprehensive collection of resources relating to invisible radiation photography but there are plans to expand the site to a range of clinical recording topics, and other potential contributors are asked to join the project. It contains a vast collection of photographs from many photographers as well as graphs, diagrams, tables and references. This paper also discusses some of the important issues surrounding the 'publication' of such a site such as currency and access versus credibility; technological obsolescence, site design and usage. PMID- 12893565 TI - The development of an improved method of photography for mole-monitoring at the University Hospital of North Durham. AB - The past decade has witnessed a dramatic increase in the number of patients presenting with pigmented lesions to general practitioners. This increase is reflected in the number of patients referred for whole body mole-monitoring photographs to the Medical Photography and Illustration Department of the University Hospital of North Durham. Through a process of qualitative and quantitative research, an improved method of whole body photographic mole monitoring was devised and implemented within this department. All aspects of the mole-monitoring service were evaluated, from the original photographic request to the photographic session and patient interaction. This process also offered the opportunity to explore the possible use of new technologies. PMID- 12893569 TI - Whither medical illustration? PMID- 12893572 TI - The mechanics of PowerPoint. PMID- 12893573 TI - Using PowerPoint. PMID- 12893574 TI - Paget's Disease of the Bone. PMID- 12893575 TI - Diabetic retinopathy screening. AB - As part of the Institute of Medical Illustrators' (IMI) scheme for continuing professional development (CPD), worksheets will be published at regular intervals in this Journal. These are designed to provide the members of IMI with a structured CPD activity that offers one way to earn credits. It is recognized that this worksheet requires some time spent obtaining and reading a number of publications. As a way of ensuring that the allocated time has been spent on the activities a number of self-assessment exercises (SAEs) have been included. Upon completion of this worksheet, you will be credited with 10 CPD points. The answers to the SAEs, along with any notes you make and other publications you find, should be kept in your CPD portfolio. PMID- 12893581 TI - Describing ethnicity in health research. AB - Commentators have criticised the terminology used for the classification of ethnic and racialised groups in health research for a number of years. The shortcomings of fixed-response categories include the reproduction of racialised categorisations, overemphasis of homogeneity within groups and contrast between them, and failure to offer terms with which people identify and which can express complex identities. The historical injustices against black and minority groups are reflected in terminology and explicitly recognised when discussing 'race' as a social construction. The exaggeration of homogeneity within groups and contrast between them is a racialising effect of fixed classifications. Self-assigned ethnic group avoids some of these difficulties by allowing multiple affiliations to be described, but introduces the costs of processing free text. The context dependent nature of individual ethnic identity makes comparison problematic. Researcher-assigned ethnicity can increase comparability and consistency but may be at odds with self-identity. The complexity of ethnicity itself and of its relationship with socio-economic group and racism makes proxy measures inevitably inadequate. If researchers continue to try to capture the complex and contextual detail of ethnicity, it may become clear that the general concept of ethnicity covers such a wide and specific range of experiences as to render it of limited use in making comparisons through time or across cultures. PMID- 12893582 TI - Researching sensitive issues and ethnicity: lessons from sexual health. AB - Researchers are generally familiar with the need to adapt their research methods in order to minimise the impact of sensitivity. However, are different considerations required for studies of ethnicity or with ethnic minority respondents? This paper provides insight into strategies used by the authors to tackle sensitivity in qualitative and quantitative research studies of sexual health and ethnicity. Drawing on examples from these studies, the paper demonstrates the relevance of Lee's model of sensitivity, and examines ways in which ethnicity may add further dimensions to our ability to understand, identify and limit sensitivity in the research process. The paper identifies techniques available to researchers for handling sensitivity in general, as well as those that may be useful for researching sensitive topics among ethnic minority populations. The paper concludes that, given the transitional and contextual nature of ethnicity and sensitivity, formulaic approaches for tackling the latter are inappropriate. Instead, strategies are best developed in collaboration with the study communities. PMID- 12893583 TI - Including culturally diverse samples in health research: a case study of an urban trial of social support. AB - OBJECTIVES: To describe the recruitment procedures used in a study of Social Support and Family Health carried out in a disadvantaged urban area of the UK in 1999-2001; to consider the impact of using inclusive recruitment procedures on the final research sample and implications for the conduct of the research and data obtained. DESIGN: Face-to-face recruitment of eligible women, using interpreters where necessary, to a randomized controlled trial of two alternative strategies for providing support to women with infants. RESULTS: Of the 1,263 women eligible to enter the trial, 731 were successfully recruited. Forty-five languages other than English were spoken by eligible women; 14% needed an interpreter for the recruitment visit, and a further 30% spoke English as a second language. Inclusive recruitment practices added significant costs, resulted in a study sample with a different social profile from the sample that would have been achieved without these, and challenged some of the assumptions underlying the model of informed consent commonly used in much health services research. CONCLUSION: Procedures can be developed for recruiting people with diverse cultural backgrounds to take part in research. This helps to address the issue of possible bias in generalizing research findings by increasing external validity, and respects the ethic that everyone should have the right to be eligible for inclusion in research. PMID- 12893584 TI - Locating research informants in a multi-ethnic community: ethnic identities, social networks and recruitment methods. AB - OBJECTIVE: Recruitment of informants can 'make or break' social research projects, yet this has received little research attention. Drawing on our recent qualitative research into health and social capital in a multi-ethnic neighbourhood in South England, this paper presents a detailed analysis of the complexities encountered in recruiting research informants who described themselves as African-Caribbean, Pakistani-Kashmiri and white English. METHODS: Three methods of recruitment were used: (1) advertisements and articles in local media, (2) institutional contacts through local voluntary organisations and (3) interpersonal contacts, referrals and snowballing. We compare and contrast the experiences of ethnically matched interviewers who conducted research amongst the three aforementioned ethnic groups. These experiences were recorded by means of lengthy interviewer 'debriefing questionnaires' that focused on factors that had served to help or hinder them in finding research participants. These questionnaires formed the basis of a discussion workshop in which the interviewers and researchers sought to identify the factors impacting on the recruitment process. RESULTS: Our findings suggest that local advertisements and media contact worked best for recruiting members of the white English community in our South English town. Interpersonal contacts were crucial in recruiting Pakistani-Kashmiri informants. Institutional contacts were the most useful way of accessing African-Caribbean individuals. CONCLUSION: We conclude that local ethnic identities and social networks produce qualitatively different responses to recruitment attempts in different communities. Such differences necessitate the employment of a range of recruitment methodologies and detailed formative research in a target community before commencing recruitment. PMID- 12893585 TI - Can findings from postal questionnaires be combined with interview results to improve the response rate among ethnic minority populations? AB - STUDY OBJECTIVE: To maximise the response rate in a community survey among ethnic minorities by combining postal questionnaires and interviews and to evaluate the validity of combining results from these different methods. DESIGN: A cross sectional community survey of a local population using postal questionnaires with interview questionnaires for non-respondents. Postal questionnaires were in English and interview questionnaires were prepared in South Asian languages. A sub-sample completed both postal and interview questionnaires. SETTING: Two general practices in Tameside, Greater Manchester, UK. PARTICIPANTS: Questionnaires were mailed to 1,267 people. People were included if they defined their ethnicity as Indian, Pakistani, Bangladeshi or a combination of these. Fifty-five people who returned postal questionnaires were also interviewed. MAIN RESULTS: Overall response rate was 75%. Comparison of questionnaire and interview responses produced values of kappa ranging from marginally below zero to one. Equivalence was greater with a shorter time between postal completion and interview and where questions were more objective. CONCLUSIONS: It is possible to achieve a good response rate for an epidemiological study among ethnic minorities by using both postal questionnaires and interviews. Care should be taken when results from these two methods are combined, since equivalence is uncertain. PMID- 12893586 TI - Cancer screening practices and attitudes: comparison of low-income women in three ethnic groups. AB - OBJECTIVE: Based on the Health Belief Model, this study investigates differences among ethnically diverse, low-income women in the USA to inform better outreach strategies to encourage participation in the Centers for Disease Control & Prevention (CDC)- sponsored breast and cervical cancer early detection program. DESIGN: Program-eligible, low-income, Hispanic, Vietnamese and Cambodian American women who were over the age of 40 volunteered to be interviewed in their first language for the study. A total of 78 women completed the interviews. RESULTS: All three samples of women were more likely to perceive barriers to having a mammogram performed compared to the non-minority normative group. Hispanic and Vietnamese women were more similar in their health beliefs and behaviors than Vietnamese and Cambodian women. CONCLUSION: This study supports other research on the barriers and health belief differences found among ethnic minority women in the USA. Further, these findings suggest that it is not advisable to collapse ethnic groups into general categories such as 'Asian' when planning cancer control strategies, as differences were found by country of origin. PMID- 12893587 TI - Preemptive use of antiretroviral drugs to prevent sexual transmission of HIV to high-risk uninfected MSM. PMID- 12893588 TI - Health status of new Americans. PMID- 12893590 TI - On reconsidering community-based health promotion. PMID- 12893592 TI - Healthy aging begins with the fetus. PMID- 12893595 TI - Partnering to advance public health: making a difference through government, community, business, and academic vocations. PMID- 12893596 TI - The road to advocacy--searching for the rainbow. AB - The essence of public health advocacy is spreading the word-spreading the word to members of one's community about ways to protect and promote health, and spreading the word to decision makers about health policies that need to be enacted. The authors profile 2 women who spread the word-one who focuses on breast cancer in the Asian American community and one who works in cooperation with churches in the fight against HIV/AIDS in African and African American communities-and discuss the importance of "creating shoulders for others to stand on" in the fight for social change. PMID- 12893597 TI - Community-based participatory research: implications for public health funding. AB - Community-based participatory research (CBPR) increasingly is being recognized by health scholars and funders as a potent approach to collaboratively studying and acting to address health disparities. Emphasizing action as a critical part of the research process, CBPR is particularly consistent with the goals of "results oriented philanthropy" and of government funders who have become discouraged by the often modest to disappointing results of more traditional research and intervention efforts in many low income communities of color. Supporters of CBPR face challenging issues in the areas of partnership capacity and readiness, time requirements, funding flexibility, and evaluation. The authors suggest strategies for addressing such issues and make a case for increasing support of CBPR as an important tool for action-oriented and community-driven public health research. PMID- 12893598 TI - The Duluth clean indoor air ordinance: problems and success in fighting the tobacco industry at the local level in the 21st century. AB - Case study methodology was used to investigate the tobacco industry's strategies to fight local tobacco control efforts in Duluth, Minn. The industry opposed the clean indoor air ordinance indirectly through allies and front groups and directly in a referendum. Health groups failed to win a strong ordinance because they framed it as a youth issue rather than a workplace issue and failed to engage the industry's economic claims. Opponents' overexploitation of weaknesses in the ordinance allowed health advocates to construct a stronger version. Health advocates should assume that the tobacco industry will oppose all local tobacco control measures indirectly, directly, or both. Clean indoor air ordinances should be framed as workplace safety issues. PMID- 12893599 TI - Public health, law, and local control: destruction of the US chemical weapons stockpile. AB - Destruction of US chemical weapons has begun at one of the 8 sites in the continental United States, was completed on Johnston Island in the Pacific Ocean, and is scheduled to begin in at least 3 other locations during the upcoming year. About 25% of the stockpile and 38% of the munitions had been destroyed as of December 31, 2002. However, the program has become controversial with regard to choice of technology, emergency management, and cost. This controversy is in large part due to efforts by some state and local governments and activist groups to play a more central role in a decision making process that was once fully controlled by the US Army. PMID- 12893600 TI - Public health's response to a changed world: September 11, biological terrorism, and the development of an environmental health tracking network. AB - Historically, the importance of public health has often been recognized during or as a result of major tragedy. The attacks that occurred in the United States in 2001 are no exception. These events have raised awareness of our vulnerability and the need for emergency preparedness, the need for a flexible and sustainable public health infrastructure, and the importance of linkages between environmental exposures and health outcomes. The authors encourage the public health community, along with policymakers, to develop a national environmental health tracking system that can improve our overall public health capacity and prepare us to investigate the critical issues of the day, whether they be emerging infectious diseases, terrorist attacks, or chronic illnesses. PMID- 12893601 TI - Innovative surveillance methods for rapid detection of disease outbreaks and bioterrorism: results of an interagency workshop on health indicator surveillance. AB - A system designed to rapidly identify an infectious disease outbreak or bioterrorism attack and provide important demographic and geographic information is lacking in most health departments nationwide. The Department of Defense Global Emerging Infections System sponsored a meeting and workshop in May 2000 in which participants discussed prototype systems and developed recommendations for new surveillance systems. The authors provide a summary of the group's findings, including expectations and recommendations for new surveillance systems. The consensus of the group was that a nationally led effort in developing health indicator surveillance methods is needed to promote effective, innovative systems. PMID- 12893602 TI - The fox guarding the chicken coop: monitoring exposure to respirable coal mine dust, 1969-2000. AB - Following passage of the Coal Mine Health and Safety Act of 1969, underground coal mine operators were required to take air samples in order to monitor compliance with the exposure limit for respirable dust, a task essential for the prevention of pneumoconiosis among coal workers. Miners objected, claiming that having the mine operators perform this task was like "having the fox guard the chicken coop." This article is a historical narrative of mining industry corruption and of efforts to reform the program of monitoring exposure to coal mine dust. Several important themes common to the practice of occupational health are illustrated; most prominently, that employers should not be expected to regulate themselves. PMID- 12893603 TI - Buried in mud, digging for gold. PMID- 12893604 TI - The condition of the working class in England. 1845. PMID- 12893605 TI - Friedrich Engels: businessman and revolutionary. PMID- 12893606 TI - Student leadership in public health advocacy: lessons learned from the hepatitis B initiative. AB - Increasing hepatitis B vaccination rates for Asian Americans and Pacific Islanders is a priority. Laws requiring vaccination prior to school enrollment have helped, yet many youths remain unvaccinated. The Hepatitis B Initiative (HBI), launched in 1997 and operated by public health and medical school students, provides free screenings and vaccinations to Boston's Asian American/Pacific Islander community, with a focus on youths. By October 2002, 997 HBI patients from Boston's Chinatown had received free hepatitis B screenings. Of these, 384 patients (39%) were deemed susceptible to the hepatitis B virus and provided with free vaccination. PMID- 12893607 TI - Should the Centers for Disease Control and Prevention's childhood lead poisoning intervention level be lowered? AB - The US Centers for Disease Control and Prevention (CDC) in 1991 chose 10 micro g/dL as an initial screening level for lead in children's blood. Current data on health risks and intervention options do not support generally lowering that level, but federal lead poisoning prevention efforts can be improved by revising the follow-up testing schedule for infants aged 1 year or less with blood lead levels of 5 micro g/dL or higher; universal education about lead exposure risks; universal administration of improved, locally validated risk-screening questionnaires; enhanced compliance with targeted screening recommendations and federal health program requirements; and development by regulatory agencies of primary prevention criteria that do not use the CDC's intervention level as a target "safe" lead exposure. PMID- 12893608 TI - Why don't we see more translation of health promotion research to practice? Rethinking the efficacy-to-effectiveness transition. AB - The gap between research and practice is well documented. We address one of the underlying reasons for this gap: the assumption that effectiveness research naturally and logically follows from successful efficacy research. These 2 research traditions have evolved different methods and values; consequently, there are inherent differences between the characteristics of a successful efficacy intervention versus those of an effectiveness one. Moderating factors that limit robustness across settings, populations, and intervention staff need to be addressed in efficacy studies, as well as in effectiveness trials. Greater attention needs to be paid to documenting intervention reach, adoption, implementation, and maintenance. Recommendations are offered to help close the gap between efficacy and effectiveness research and to guide evaluation and possible adoption of new programs. PMID- 12893609 TI - Barriers to colorectal cancer screening with fecal occult blood testing in a predominantly minority urban population: a qualitative study. PMID- 12893610 TI - Sociocultural correlates of breast cancer knowledge and screening in urban African American women. PMID- 12893611 TI - The impact of rubella immunization on the serological status of women of childbearing age: a retrospective longitudinal study in Melbourne, Australia. PMID- 12893612 TI - Evaluation of the impact of the baby-friendly hospital initiative on rates of breastfeeding. PMID- 12893613 TI - Prevalence of diagnosed diabetes and related risk factors: Japanese adults in Westchester County, New York. PMID- 12893614 TI - Descriptions of Web sites in search listings: a potential obstacle to informed choice of health information. PMID- 12893615 TI - Declining trends in work-related morbidity and disability, 1993-1998: a comparison of survey estimates and compensation insurance claims. AB - OBJECTIVES: This study compared trends in the incidence of work-related morbidity and disability across 3 sources of surveillance data in a Canadian province. METHODS: Time series estimates of workplace injuries and work-related disability based on 2 panel surveys in the province of Ontario, Canada, for the period 1993 1998 were compared with rates of work-related injury and illness compensation claims during the same period. RESULTS: Lost-time compensation claims declined by 28.8% over this 6-year period. The incidence of self-reported work-related injury declined by 28.2%, and the self-reported incidence of work absence for work related causes declined by 32.2%. CONCLUSIONS: Parallel reductions in work related morbidity were seen in 3 independent data sources. These results support an interpretation that there has been an important reduction in injury risk in Ontario workplaces over the past decade. PMID- 12893616 TI - Income inequality, household income, and health status in Canada: a prospective cohort study. AB - OBJECTIVES: This study sought to determine whether income inequality, household income, and their interaction are associated with health status. METHODS: Income inequality and area income measures were linked to data on household income and individual characteristics from the 1994 Canadian National Population Health Survey and to data on self-reported health status from the 1994, 1996, and 1998 survey waves. RESULTS: Income inequality was not associated with health status. Low household income was consistently associated with poor health. The combination of low household income and residence in a metropolitan area with less income inequality was associated with poorer health status than was residence in an area with more income inequality. CONCLUSIONS: Household income, but not income inequality, appears to explain some of the differences in health status among Canadians. PMID- 12893617 TI - Income inequality, illiteracy rate, and life expectancy in Brazil. AB - OBJECTIVES: The link between income disparities and health has been studied mostly in developed nations. This study assesses the relationship between income disparities and life expectancy in Brazil and measures the impact of illiteracy rates on the association. METHODS: The units of analysis (n = 27) are all the Brazilian states and the federal capital. Simple and multiple linear regressions were performed to measure the association between income disparity, measured by the Gini coefficient, gross domestic product (GDP) per capita, and illiteracy rate. Data came from publicly available sources at the Brazilian Ministry of Health and the Brazilian Institute of Geography and Statistics. RESULTS: Income disparities and illiteracy rates were negatively associated with life expectancy in Brazil. GDP per capita was positively associated with life expectancy. The inclusion of illiteracy rates in the regression model removed the effect of income disparities. CONCLUSIONS: Illiteracy rate is strongly associated with life expectancy in Brazil. This finding is in accord with reports from the United States and has implications for health policy and planning for both developed and developing countries. PMID- 12893618 TI - A Medicaid population's use of physicians' offices for dental problems. AB - OBJECTIVES: This study evaluated how the elimination of Medicaid reimbursement to dentists for the treatment of adult dental problems affected patients' visits to physicians. METHODS: Data tapes describing physicians' claims for adult Medicaid patients were obtained from the Maryland Medicaid Management Information System. The database contains information on all claims made to Maryland Medicaid, including date, provider, International Classification of Diseases, Ninth Revision, Clinical Modification Manual code, and payments. RESULTS: A total of 5334 individuals made physician's office claims related to dental problems sometime during the 4-year study period. The rate of dental-related claims by physicians decreased by 8% after the policy change. CONCLUSIONS: Visits to physicians' offices decreased even though an increase might have been expected because of the elimination of access to dentists in private practice. Patients might have assumed that if visits to dentists would no longer be paid for, neither would visits to physicians' offices. PMID- 12893619 TI - Trends in risk factors for lifestyle-related diseases by socioeconomic position in Geneva, Switzerland, 1993-2000: health inequalities persist. AB - OBJECTIVES: We report on trends in risk factors for lifestyle-related diseases among socioeconomic position (SEP) groups. METHODS: We continuously surveyed the adult population of Geneva, Switzerland, for 8 years (1993-2000) with independent, cross-sectional surveys of representative samples (4207 men and 3987 women aged 35-74 years). Age-adjusted linear regression slopes estimated annual risk factor trends. Interaction terms were tested for trend differences between SEP groups. RESULTS: Overall, low-SEP persons had the worst risk factor profiles. Eight-year trends indicate that (1) number of pack-years smoked decreased by half a pack-year among high-SEP female current smokers only; (2) obesity prevalence more than doubled from 5% to 11% among high-SEP men only; (3) systolic and diastolic blood pressures decreased similarly in all SEP groups; (4) unsaturated to-saturated dietary fat ratio declined in the low-SEP group only; and (5) physical inactivity and current/former cigarette smoking prevalences remained unchanged in all SEP groups. CONCLUSIONS: Smoking, obesity, high blood pressure, and physical inactivity are more prevalent among low-SEP persons. Most socioeconomic risk factor differences remained stable in the 1990s. Thus, social inequalities in chronic disease morbidity and mortality will persist in the next decades. PMID- 12893620 TI - Childhood pesticide exposures on the Texas-Mexico border: clinical manifestations and poison center use. AB - OBJECTIVES: The purpose of this study was to describe differences in childhood pesticide exposures between counties on the Texas-Mexico border and nonborder counties. METHOD: The authors reviewed all pesticide exposures among children younger than 6 years reported to the South Texas Poison Center during 1997 through 2000. RESULTS: Nonborder counties had twice the reported exposure rate of border counties. Parents of border children were significantly less likely to contact the poison center after an exposure and more likely to have their children evaluated in a health care facility. CONCLUSIONS: Increasing residents' awareness of the poison center and identifying potential barriers to its use among residents of Texas-Mexico border communities may prevent unnecessary visits to health care facilities. PMID- 12893621 TI - The relationship of health insurance to the diagnosis and management of asthma and respiratory problems in children in a predominantly Hispanic urban community. AB - OBJECTIVES: As part of an asthma screening study, we evaluated the relationship of health care insurance coverage to the diagnosis and treatment of elementary school children for asthma and related respiratory problems from 1998 through 2001. METHODS: A bilingual questionnaire assessing health care coverage, asthma diagnosis, respiratory symptoms, and use of medications was distributed to parents of 6235 public and private school children in grades 2 through 5 in Passaic, NJ. RESULTS: Responses for 4380 children (70%) revealed disparities in health care coverage and asthma diagnosis among racial and ethnic groups. Mexican and Dominican children had significant increases in health care coverage over the 4 years. CONCLUSIONS: The percentage of children with health insurance grew from 67% in 1998 to 81% in 2001, and the increase was related to NJ KidCare. Diagnosis of asthma and treatment were related to health care coverage. PMID- 12893622 TI - Nondaily smokers: who are they? AB - OBJECTIVE: We sought to understand who constitutes the sizable population of nondaily, or some-day (SD), smokers. METHODS: We analyzed descriptive statistics and regression results using the 1998-1999 Current Population Survey Tobacco Use Supplement to determine the prevalence of SD smokers, their sociodemographic characteristics, and the smoking patterns and histories of groups differentiated by the length and stability of their SD smoking. RESULTS: SD smokers make up 19.2% of all current smokers. Among SD smokers, 44.6% have smoked less than daily for at least 1 year, no more than 14.4% are just starting to smoke, and the rest are likely in transition. Overall, SD smokers smoked a mean of 102 cigarettes per month (compared to 566.4 for daily smokers), on an average of 14.5 days out of the past 30. CONCLUSIONS: SD smokers make up a substantial segment of the smoking population. They are not just beginning to smoke nor trying to quit. Many have developed a long-standing pattern of nondaily smoking, smoking relatively few cigarettes on the days when they do smoke. They are not substantially younger than daily smokers, as one might expect. PMID- 12893623 TI - Primary care attributes and care for depression among low-income African American women. AB - OBJECTIVES: We examined the association between attributes of primary care providers and care for depression, from a patients' perspective, among a sample of predominantly low-income African American women. METHODS: Computer-assisted telephone interviews were conducted among a population-based sample of 1202 women residing in Washington, DC. RESULTS: Respondents whose primary care physicians provided more comprehensive medical services were more likely to be asked about and treated for depressive symptoms than women whose providers were less medically comprehensive. Women who rated their providers as having more respect for them also were more likely to be asked about and treated for depression. CONCLUSIONS: More comprehensive primary care delivery and a physician-patient relationship focused on mutual respect are associated with greater rates of physician inquiry about and treatment for depression among vulnerable women. PMID- 12893624 TI - A comparison of national estimates from the National Health Interview Survey and the Behavioral Risk Factor Surveillance System. AB - OBJECTIVES: The purpose of this study was to compare national estimates from the National Health Interview Survey (NHIS) and the Behavioral Risk Factor Surveillance System (BRFSS). METHODS: The authors compared data from the 2 surveys on smoking, height, weight, body mass index, diabetes, hypertension, immunization, lack of insurance coverage, cost as a barrier to medical care, and health status. RESULTS: Overall national estimates were similar for 13 of the 14 measures examined. Small differences according to demographic characteristics were found for height and body mass index, with larger differences for health status. CONCLUSIONS: Although estimates differed within subgroups, the BRFSS provided national estimates comparable to those of the NHIS. BRFSS national data could provide rapidly available information to guide national policy and program decisions. PMID- 12893625 TI - Portrayals of overweight and obese individuals on commercial television. AB - OBJECTIVES: This study examined the distribution and individual characteristics of body types on prime-time television. METHODS: Five episodes of each of the 10 top-rated prime-time fictional programs on 6 broadcast networks during the 1999 2000 season were quantitatively analyzed. RESULTS: Of 1018 major television characters, 14% of females and 24% of males were overweight or obese, less than half their percentages in the general population. Overweight and obese females were less likely to be considered attractive, to interact with romantic partners, or to display physical affection. Overweight and obese males were less likely to interact with romantic partners and friends or to talk about dating and were more likely to be shown eating. CONCLUSIONS: Overweight and obese television characters are associated with specific negative characteristics. PMID- 12893626 TI - A neurokinin 1 receptor antagonist reduces an ongoing ileal pouch inflammation and the response to a subsequent inflammatory stimulus. AB - Ileal pouch-anal anastomosis (IPAA) is an excellent surgical option for patients with chronic ulcerative colitis (CUC) requiring colectomy; however, persistent episodes of ileal pouch inflammation, or pouchitis, may result in debilitating postoperative complications. Because considerable evidence implicates substance P (SP) as an inflammatory mediator of CUC, we investigated whether SP participates in the pathophysiology of pouchitis. With the use of a rat model of IPAA that we developed, we showed that ileal pouch MPO levels and neurokinin 1 receptor (NK 1R) protein expression by Western blot analysis were significantly elevated 28 days after IPAA surgery. In situ hybridization and immunohistochemistry showed that the increase in NK-1R protein expression was localized to the lamina propria and epithelia of pouch ileum. The intraperitoneal administration of the NK-1R antagonist (NK-1RA) CJ-12,255 for 4 days, starting on day 28, was effective in reducing MPO levels. Starting on day 28, animals with IPAA were given 5% dextran sulfate sodium (DSS) in their drinking water for 4 days, which caused histological and physical signs of clinical pouchitis concomitant with significant increases in ileal pouch MPO concentrations as well as NK-1R protein expression by Western blot analysis. In situ hybridization and immunohistochemistry showed that the increase in NK-1R protein expression was especially evident in crypt epithelia of pouch ileum. When the NK-1RA was administered 1 day before starting DSS and continued for the duration of DSS administration, the physical signs of clinical pouchitis and the rise in MPO were prevented. These data implicate SP in the pathophysiology of pouchitis and suggest that NK-1RA may be of therapeutic value in the management of clinical pouchitis. PMID- 12893627 TI - Cannabinoid receptor type 1 modulates excitatory and inhibitory neurotransmission in mouse colon. AB - The effects of cannabinoid receptor agonists and antagonists on smooth muscle resting membrane potentials and on membrane potentials following electrical neuronal stimulation in a myenteric neuron/smooth muscle preparation of wild-type and cannabinoid receptor type 1 (CB1)-deficient mice were investigated in vitro. Double staining for CB1 and nitric oxide synthase (neuronal) was performed to identify the myenteric CB1-expressing neurons. Focal electrical stimulation of the myenteric plexus induced a fast (f) excitatory junction potential (EJP) followed by a fast and a slow (s) inhibitory junction potential (IJP). Treatment of wild-type mice with the endogenous CB1 receptor agonist anandamide reduced EJP while not affecting fIJP and sIJP. EJP was significantly higher in CB1-deficient mice than in wild-type littermate controls, and anandamide induced no effects in CB1-deficient mice. N-arachidonoyl ethanolamide (anandamide), R-[2,3-dihydro-5 methyl-3-(4-morpholinylmethyl)pyrrolo[1,2,3,-de]- 1,4-benzoxazin-6-yl]-1 naphtalenylmethanone, a synthetic CB1 receptor agonist, nearly abolished EJP and significantly reduced the fIJP in wild-type mice. N-piperidino-5-(4-chlorophenyl) 1-(2,4-dichlorophenyl)-4-methyl-3-pyrazole-caroxamide (SR141716A), a CB1-specific receptor antagonist, was able to reverse the agonist effects induced in wild-type mice. SR141716A, when given alone, significantly increased EJP in wild-type mice without affecting IJP in wild-type and EJP in CB1-deficient mice. Interestingly, SR141716A reduced fIJP in CB1-deficient mice. In the mouse colon, nitrergic myenteric neurons do not express CB1, implying that CB1 is expressed in cholinergic neurons, which is in line with the functional data. Finally, excitatory and inhibitory neurotransmission in the mouse colon is modulated by activation of CB1 receptors. The significant increase in EJP in CB1-deficient mice strongly suggests a physiological involvement of CB1 in excitatory cholinergic neurotransmission. PMID- 12893628 TI - Sodium current in human intestinal interstitial cells of Cajal. AB - Interstitial cells of Cajal (ICC) generate the electrical slow wave required for normal gastrointestinal motility. The ionic conductances expressed in human intestinal ICC are unknown. The aim of this study was to determine expression of a Na+ current in human intestinal ICC and to determine the effects of the Na+ current on the slow wave. Visually identified, freshly dissociated, single ICC were verified by the presence of c-kit mRNA by using single-cell RT-PCR. Standard whole cell currents were recorded from patch-clamped ICC held at -100 mV between pulse protocols. A Na+ current was identified in human intestinal ICC. The current activated at -55 mV and peaked at -30 mV. Extracellular N-methyl-d glucamine abolished and QX-314 (500 microM) blocked the Na+ current, but nifedipine and Ni2+ did not. The Na+ current was activated by shear stress. Single-cell RT-PCR detected mRNA for the Na+ alpha-subunit SCN5A in single human intestinal ICC. Lidocaine (200 microm) and QX-314 (500 microM) decreased slow wave frequency, and stretch increased slow wave frequency. A mechanosensitive Na+ channel current is present in human intestinal ICC and appears to play a role in the control of intestinal motor function. PMID- 12893629 TI - Regulation of intestinal NaPi-IIb cotransporter gene expression by estrogen. AB - The current experiments were designed to study the effect of beta-estradiol on type IIb sodium-coupled phosphate (NaPi-IIb) cotransporter gene expression. Uptake studies with intestinal brush-border membrane vesicles (BBMV) showed that estrogen treatment increased sodium-dependent phosphate absorption by approximately 45% in rat intestine. Northern blot analysis indicated that NaPi IIb mRNA expression was increased by approximately 50% after estrogen treatment. Western blot analysis also detected an increase in BBMV NaPi-IIb protein expression in estrogen-treated rats. In human intestinal Caco-2 cells, NaPi-IIb mRNA abundance was increased approximately 60% after estrogen treatment, and this increase could be abolished by inhibition of gene transcription. Transfection studies with human NaPi-IIb promoter reporter constructs showed that the promoter was responsive to estrogen treatment. These studies demonstrate for the first time that estrogen stimulates intestinal sodium-dependent phosphate absorption in female rats. This stimulation is associated with increased NaPi-IIb mRNA and protein expression. Thus the effect of estrogen on intestinal Pi absorption may be partially due to activation of NaPi-IIb gene transcription. PMID- 12893630 TI - Identification of an apical Cl-/HCO3- exchanger in gastric surface mucous and duodenal villus cells. AB - The molecular identity of the apical HCO3(-)-secreting transporter in gastric mucous cells remains unknown despite its essential role in preventing injury and ulcer by gastric acid. Here we report the identification of a Cl-/HCO3- exchanger that is located on apical membranes of gastric surface epithelial cells. RT-PCR studies of mouse gastrointestinal tract mRNAs demonstrated that this transporter, known as anion exchanger isoform 4 (AE4), is expressed in both stomach and duodenum. Northern blot analysis of RNA from purified stomach epithelial cells indicated that AE4 is expressed at higher levels in mucous cells than in parietal cells. Immunoblotting experiments identified AE4 as a approximately 110- to 120 kDa protein in membranes from stomach epithelium and apical membranes from duodenum. Immunocytochemical staining demonstrated that AE4 is expressed in apical membranes of surface cells in both mouse and rabbit stomach and duodenum. Functional studies in oocytes indicated that AE4 functions as a Cl-/HCO3- exchanger. These data show that AE4 is an apical Cl-/HCO3- exchanger in gastric mucous cells and duodenal villus cells. On the basis of its function and location, we propose that AE4 may play an important role in mucosal protection. PMID- 12893631 TI - Chelerythrine stimulates GSH transport by rat Mrp2 (Abcc2) expressed in canine kidney cells. AB - Rat multidrug resistant protein 2 (Mrp2; Abcc2), an ATP-driven pump located on the canalicular domain of hepatocytes, exports glutathione S-conjugates (GS-X) and GSH among its wide variety of substrates. Previous studies have shown that chelerythrine (CHEL), a quaternary benzophenanthridine cation, reacts with GSH to form a reversible adduct under physiological conditions. Here we report that CHEL can strongly stimulate GSH efflux by Mrp2, when it is constitutively expressed in polarized canine kidney cells, thereby leading to the depletion of cellular GSH. Transepithelial transport experiments indicate that Mrp2 transports GSH and CHEL with a 1:1 stoichiometry, which can be readily inhibited by GS-bimane, a GS-X substrate for Mrp2. Moreover, CHEL can block Mrp2-mediated leukotriene C4 uptake by membrane vesicles with an IC50 approximately 100 microM in the presence of GSH, but not S-methyl GSH or ophthalmic acid. Thus the thiol group of GSH is required for inhibition of Mrp2 in the presence of CHEL. Our results suggest that CHEL stimulates GSH efflux by forming a reversible GS-CHEL adduct, which is transported by Mrp2 and dissociates extracellularly. PMID- 12893632 TI - Contribution of capsaicin-sensitive sensory neurons to stress-induced increases in gastric tissue levels of prostaglandins in rats. AB - We examined whether capsaicin-sensitive sensory neurons might be involved in the increase in the gastric tissue level of prostaglandins, thereby contributing to the reduction of water immersion restraint stress (WIR)-induced gastric mucosal injury in rats. Gastric tissue levels of calcitonin gene-related peptide (CGRP), 6-keto-PGF1alpha, and PGE2 were transiently increased 30 min after WIR. These increases were significantly inhibited by subcutaneous injection of capsazepine (CPZ), a vanilloid receptor antagonist, and by functional denervation of capsaicin-sensitive sensory neurons induced by the administration of high-dose capsaicin. The administration of capsaicin (orally) and CGRP (intravenously) significantly enhanced the WIR-induced increases in the gastric tissue level of prostaglandins 30 min after WIR, whereas CGRP-(8-37), a CGRP receptor antagonist, significantly inhibited them. Pretreatment with Nomega-nitro-L-arginine methyl ester (L-NAME), a nonselective inhibitor of nitric oxide (NO) synthase (NOS), and that with indomethacin inhibited the WIR-induced increases in gastric tissue levels of prostaglandins, whereas either pretreatment with aminoguanidine (AG), a selective inhibitor of the inducible form of NOS, or that with NS-398, a selective inhibitor of cyclooxygenase (COX)-2, did not affect them. CPZ, the functional denervation of capsaicin-sensitive sensory neurons, and CGRP-(8-37) significantly increased gastric MPO activity and exacerbated the WIR-induced gastric mucosal injury in rats subjected to 4-h WIR. The administration of capsaicin and CGRP significantly increased the gastric tissue levels of prostaglandins and inhibited both the WIR-induced increases in gastric MPO activity and gastric mucosal injury 8 h after WIR. These effects induced by capsaicin and CGRP were inhibited by pretreatment with L-NAME and indomethacin but not by pretreatment with AG and NS-398. These observations strongly suggest that capsaicin-sensitive sensory neurons might release CGRP, thereby increasing the gastric tissue levels of PGI2 and PGE2 by activating COX-1 through activation of the constitutive form of NOS in rats subjected to WIR. Such activation of capsaicin-sensitive sensory neurons might contribute to the reduction of WIR induced gastric mucosal injury mainly by inhibiting neutrophil activation. PMID- 12893633 TI - CaM kinase II regulation of CRHSP-28 phosphorylation in cultured mucosal T84 cells. AB - Ca(2+)-regulated heat-stable protein of 28 kDa (CRHSP-28; a member of the tumor protein D52 family) is highly expressed in exocrine glands and was shown to regulate digestive enzyme secretion from pancreatic acinar cells. We found CRHSP 28 highly expressed in cultured mucosal secretory T84 cells, consistent with an important regulatory role in apical membrane trafficking. Stimulation of cells with carbachol (CCh) induced rapid, concentration-dependent phosphorylation of CRHSP-28 on at least two serine residues. Isoelectric focusing and immunoblotting were used to characterize cellular mechanisms governing CRHSP-28 phosphorylation. Phosphorylation depends on elevated cellular Ca2+, being maximally induced by ionomycin and thapsigargin and fully inhibited by BAPTAAM. In vitro phosphorylation of recombinant CRHSP-28 was 10-fold greater by casein kinase II (CKII) than Ca2+/calmodulin-dependent protein kinase II (CaMKII). However, phosphopeptide mapping studies demonstrated that CaMKII induced an identical phosphopeptide profile to endogenous CRHSP-28 immunoprecipitated from T84 cells. Although calmodulin antagonists had no effect on CCh-stimulated phosphorylation, disruption of actin filaments by cytochalasin D inhibited phosphorylation by 50%. Confocal microscopy indicated that CRHSP-28 is expressed in perinuclear regions of cells and accumulates immediately below the apical membrane of polarized monolayers following CCh stimulation. CaMKII was also localized to the subapical cytoplasm and was clearly displaced following actin filament disruption. These data suggest that CRHSP-28 phosphorylation is regulated by a CaMKII-like enzyme and likely involves a translocation of the protein within the apical cytoplasm of epithelial cells. PMID- 12893634 TI - Carotid baroreflex function during and following voluntary apnea in humans. AB - Muscle sympathetic nerve activity (MSNA) and arterial pressure increase concomitantly during apnea, suggesting a possible overriding of arterial baroreflex inhibitory input to sympathoregulatory centers by apnea-induced excitatory mechanisms. Apnea termination is accompanied by strong sympathoinhibition while arterial pressure remains elevated. Therefore, we hypothesized that the sensitivity of carotid baroreflex control of MSNA would decrease during apnea and return upon apnea termination. MSNA and heart rate responses to -60-Torr neck suction (NS) were evaluated during baseline and throughout apnea. Responses to +30-Torr neck pressure (NP) were evaluated during baseline and throughout 1 min postapnea. Apnea did not affect the sympathoinhibitory or bradycardic response to NS (P > 0.05); however, whereas the cardiac response to NP was maintained postapnea, the sympathoexcitatory response was reduced for 50 s (P < 0.05). These data demonstrate that the sensitivity of carotid baroreflex control of MSNA is not attenuated during apnea. We propose a transient rightward and upward resetting of the carotid baroreflex-MSNA function curve during apnea and that return of the function curve to, or more likely beyond, baseline (i.e., a downward and leftward shift) upon apnea termination may importantly contribute to the reduced sympathoexcitatory response to NP. PMID- 12893635 TI - Longitudinal strain quantitates regional right ventricular contractile function. AB - The assessment of contractile function of the right ventricle (RV) is an important clinical issue, but this remains difficult because of its complex anatomy and structure. We thought to investigate whether new Doppler-derived myocardial deformation indexes may quantify regional contractile RV function during varying loading conditions. In nine pigs, ultrasonic crystals were inserted longitudinally in the RV inflow and outflow tracts to assess regional contractile function. The same RV segments and the interventricular septum were imaged using apical echocardiographic views. Regional function was assessed using two parameters: 1) systolic strain (SS), representing the relative magnitude of segmental systolic shortening; and 2) its temporal derivative, peak systolic strain rate (SR), i.e., the maximal velocity of segmental shortening. Data were acquired at baseline and during partial pulmonary artery constriction (PAC) and inferior vena cava occlusion (IVCO). SS decreased significantly after PAC and IVCO in both the inflow and outflow tracts but only during IVCO in the septum. SR was less sensitive to loading variations in all segments. A significant correlation was found between SS values derived from sonomicrometry and myocardial Doppler in RV segments (r = 0.84, P < 0.001). Thus regional strain and SR provide complementary information on the heterogeneous RV contractile function and can be accurately and noninvasively quantified using Doppler myocardial imaging. PMID- 12893636 TI - Thrombin increases permeability only in venules exposed to inflammatory conditions. AB - Thrombin is widely used to stimulate a variety of responses in cultured endothelial cell monolayers as a model of acute vascular endothelial response to inflammatory mediators. However, preliminary results indicated that rat mesenteric venules did not respond acutely to thrombin. We tested the hypothesis that rat venules would respond to thrombin 24 h after prior injury by microperfusion. Vessel responsiveness was measured as hydraulic conductivity (Lp). When venules were exposed to rat thrombin (10 U/ml) within 2 h of initial perfusion with vehicle control, there was no increase in Lp of any vessel from a mean baseline of 1.2 +/- 0.2 x 10(-7) cm.s-1.cmH2O-1. In contrast, when perfused with thrombin at 25-27 h after initial perfusion, every venule responded to thrombin with a transient increase in Lp. The mean peak Lp on day 2 in response to thrombin was 24 +/- 4.2 x 10(-7) cm.s-1.cmH2O-1. Our results suggest that prior endothelial injury modifies the endothelial cell phenotype and alters the response of endothelial cells to thrombin after 24 h. Phenotypic plasticity of endothelial cells may play a key role in the regulation of permeability of some endothelial cells in culture and in intact venules, where localized leaky sites may form where there had been a previous inflammatory response. PMID- 12893637 TI - Synchronization of ventricular fibrillation with real-time feedback pacing: implication to low-energy defibrillation. AB - Wavefront synchronization is an important aspect preceding the termination of ventricular fibrillation (VF). We evaluated the defibrillation efficacy of a novel multisite pacing algorithm using optical recording-guided synchronized pacing (SyncP) in the excitable gaps. We compared the effects of SyncP with traditional overdrive pacing (ODP) at 90% of the VF cycle length (VFCL) and high frequency pacing (HFP; 43-215 Hz) on spontaneous VF termination in isolated rabbit hearts. For SyncP, the pacing current was triggered by the activation of a reference site and was delivered when the optical potential of the pacing site was in an excitable gap. We measured VFCL and the spatial dispersion of VFCL (SDCL) from five points (3 points in the paced area and 2 points in the nonpaced area) and the distribution of phase singularities during the prepacing, pacing, and postpacing periods. The results showed that 1) the VF termination rate of SyncP (16.0%, n = 106) was higher than that of ODP (2.1%, n = 48, P < 0.01) or HFP (1.6%, n = 129, P < 0.0001); 2) energy consumption for SyncP (7.6 +/- 9.3 mJ) was significantly lower than that of ODP (14.0 +/- 14.8 mJ, P < 0.0001); and 3) SyncP, but not ODP or HFP, decreased SDCL in the paced area during the pacing (P < 0.01) and postpacing (P < 0.05) periods compared with the prepacing period. We conclude that SyncP is effective in inducing wavefront synchronization and is more effective at facilitating spontaneous VF termination than non-SyncP. PMID- 12893638 TI - Behavior of ectopic surface: effects of beta-adrenergic stimulation and uncoupling. AB - By using both experimental and theoretical means, we have addressed the progression of ectopic activity from individual cardiac cells to a multicellular two-dimensional network. Experimental conditions that favor ectopic activity have been created by local perfusion of a small area of cardiomyocyte network (I-zone) with an isoproterenol-heptanol containing solution. The application of this solution initially slowed down and then fully blocked wave propagation inside the I-zone. After a brief lag period, ectopically active cells appeared in the I zone, followed by evolution of the ectopic clusters into slowly propagating waves. The changing pattern of colliding and expanding ectopic waves confined to the I-zone persisted for as long as the isoproterenol-heptanol environment was present. On restoration of the control environment, the ectopic waves from the I zone broke out into the surrounding network causing arrhythmias. The observed sequence of events was also modeled by FitzHugh-Nagumo equations and included a cell's arrangement of two adjacent square regions of 20 x 20 cells. The control zone consisted of well-connected, excitable cells, and the I-zone was made of weakly coupled cells (heptanol effect), which became spontaneously active as time evolved (isoproterenol effect). The dynamic events in the system have been studied numerically with the use of a finite difference method. Together, our experimental and computational data have revealed that the combination of low coupling, increased excitability, and spatial heterogeneity can lead to the development of ectopic waves confined to the injured network. This transient condition appears to serve as an essential step for the ectopic activity to "mature" before escaping into the surrounding control network. PMID- 12893639 TI - Effects of coupled pacing on cardiac performance during acute atrial tachycardia and fibrillation: an old therapy revisited for a new reason. AB - Atrial tachycardia (AT) and fibrillation (AF) result in rapid ventricular rates that are detrimental to optimal cardiac function. The purpose of this study was to determine whether the application of a coupled pacing (CP) regimen would improve ventricular function by decreasing the ventricular rate of mechanical contractions (VRMCs). We simulated AT by pacing either atrium at a rate that resulted in a rapid but regular ventricular rate in seven anesthetized dogs. AF was induced by increasing the atrial pacing rate until atrial activation did not follow the pacing. After the induction of either AT or AF, we applied CP after each intrinsic ventricular activation. We measured the VRMCs and left ventricular (LV) pressures and volumes via a pressure-conductance catheter. The marked reductions in VRMCs during CP resulted in increases in LV end-diastolic volume. The CP resulted in virtually no mechanical contractions, whereas the strength of contractions from the normal electrical activation increased. The increases in the positive LV rate of pressure development over time and LV ejection fraction during CP were the result of postextrasystolic potentiation. The average stroke work (area of the pressure-volume loops) increased as a result of CP during both AT and AF. Despite the large increases in stroke volume (approximately 2x) during CP, the changes in cardiac output were moderate because the VRMCs markedly decreased (approximately 1/2). We conclude that CP therapy may be a viable therapy for slowing the heart rate and improving cardiac performance in patients with AT and AF. PMID- 12893640 TI - Reactive oxygen species induce reversible PECAM-1 tyrosine phosphorylation and SHP-2 binding. AB - Platelet endothelial cell adhesion molecule-1 (PECAM-1, CD31) functions to control the activation and survival of the cells on which it is expressed. Many of the regulatory functions of PECAM-1 are dependent on its tyrosine phosphorylation and subsequent recruitment of the Src homology (SH2) domain containing protein tyrosine phosphatase SHP-2. The recent demonstration that PECAM-1 tyrosine phosphorylation occurs in cells exposed to the reactive oxygen species hydrogen peroxide (H2O2) suggested that this form of oxidative stress may also support PECAM-1/SHP-2 complex formation. In the present study, we show that PECAM-1 tyrosine phosphorylation in response to exposure of cells to H2O2 is reversible, involves a shift in the balance between kinase and phosphatase activities, and supports binding of SHP-2 and recruitment of this phosphatase to cell-cell borders. We speculate, however, that the unique ability of H2O2 to reversibly oxidize the reactive site cysteine residues of protein tyrosine phosphatases may result in transient inactivation of the SHP-2 that is bound to PECAM-1 under these conditions. Finally, we provide evidence that PECAM-1 tyrosine phosphorylation and SHP-2 binding in endothelial cells requires exposure to an "oxidative burst" of H2O2, but that exposure of these cells to sufficiently high concentrations of H2O2 for a sufficiently long period of time abrogates binding of SHP-2 to tyrosine-phosphorylated PECAM-1. These findings support a role for PECAM-1 as a sensor of oxidative stress, perhaps most importantly during the process of inflammation. PMID- 12893641 TI - Glutamate causes a loss in human cerebral endothelial barrier integrity through activation of NMDA receptor. AB - l-Glutamate is a major excitatory neurotransmitter that binds ionotropic and metabotropic glutamate receptors. Cerebral endothelial cells from many species have been shown to express several forms of glutamate receptors; however, human cerebral endothelial cells have not been shown to express either the N-methyl-D aspartate (NMDA) receptor message or protein. This study provides evidence that human cerebral endothelial cells express the message and protein for NMDA receptors. Human cerebral endothelial cell monolayer electrical resistance changes in response to glutamate receptor agonists, antagonists, and second message blockers were tested. RT-PCR and Western blot analysis were used to demonstrate the presence of the NMDA receptor. Glutamate and NMDA (1 mM) caused a significant decrease in electrical resistance compared with sham control at 2 h postexposure; this response could be blocked significantly by MK-801 (an NMDA antagonist), 8-(N,N-diethylamino)-n-octyl-3,4,5-trimethyoxybenzoate (an intracellular Ca2+ antagonist), and N-acetyl-L-cystein (an antioxidant). Trans(+/ )-1-amino-1,3-cyclopentanedicarboxylic acid, a metabotropic receptor agonist (1 mM), did not significantly decrease electrical resistance. Our results are consistent with a model where glutamate, at excitotoxic levels, may lead to a breakdown in the blood brain barrier via activation of NMDA receptors. PMID- 12893642 TI - Estimation of capillary density in human skeletal muscle based on maximal oxygen consumption rates. AB - A previously developed Krogh-type theoretical model was used to estimate capillary density in human skeletal muscle based on published measurements of oxygen consumption, arterial partial pressure of oxygen, and blood flow during maximal exercise. The model assumes that oxygen consumption in maximal exercise is limited by the ability of capillaries to deliver oxygen to tissue and is therefore strongly dependent on capillary density, defined as the number of capillaries per unit cross-sectional area of muscle. Based on an analysis of oxygen transport processes occurring at the microvascular level, the model allows estimation of the minimum number of straight, evenly spaced capillaries required to achieve a given oxygen consumption rate. Estimated capillary density values were determined from measurements of maximal oxygen consumption during knee extensor exercise and during whole body cycling, and they range from 459 to 1,468 capillaries/mm2. Measured capillary densities, obtained with either histochemical staining techniques or electron microscopy on quadriceps muscle biopsies from healthy subjects, are generally lower, ranging from 123 to 515 capillaries/mm2. This discrepancy is partly accounted for by the fact that capillary density decreases with muscle contraction and muscle biopsy samples typically are strongly contracted. The results imply that estimates of maximal oxygen transport rates based on capillary density values obtained from biopsy samples do not fully reflect the oxygen transport capacity of the capillaries in skeletal muscle. PMID- 12893643 TI - Persistent activation of nuclear factor-kappaB signaling pathway in severe uncontrolled asthma. AB - The transcription factor nuclear factor-kappaB (NF-kappaB) is inactive when bound to its inhibitory protein IkappaBalpha. On cell stimulation with inflammatory signals, IkappaBalpha is phosphorylated by IkappaB kinases and subsequently degraded. Freed NF-kappaB then induces expression of cytokines such as granulocyte-macrophage colony-stimulating factor, interleukin-8, and regulated upon activation, normal T cell expressed and secreted. These mediators are overexpressed in asthma and are downregulated by glucocorticoids through NF kappaB activity repression. However, high levels of granulocyte-macrophage colony stimulating factor, interleukin-8, and regulated upon activation, normal T cell expressed and presumably secreted are released by peripheral blood mononuclear cells isolated from patients with severe asthma despite continuous systemic glucocorticoid treatment. We report that these mediators are markedly decreased by pyrrolidinedithiocarbamate, an inhibitor of NF-kappaB activation. To further characterize the persistent NF-kappaB activation in severe asthma, we analyzed the expression of various components of this activation pathway in healthy subjects and in asthmatics with mild controlled, and moderate and severe uncontrolled disease. We found high amounts of phosphorylated IkappaBalpha characterizing the three asthmatic groups. Western blot analyses indicated that in peripheral blood mononuclear cells the IkappaB kinase beta and p65 levels were greater in moderate and severe asthmatics than in normal subjects. Electrophoretic mobility shift assay and immunocytochemistry showed a greater activation status of p65 in severe asthmatics. Our data suggest that exaggerated NF-kappaB activation perpetuates inflammatory mediators production in severe asthma. PMID- 12893644 TI - Aerosol delivery and modern mechanical ventilation: in vitro/in vivo evaluation. AB - Aerosol delivery via a mechanical ventilator remains unregulated with no standards for drug delivery to intubated patients. Bench models predicting drug delivery have not been validated in vivo. For modern ventilator designs, we chose to identify, on the bench, the most important variables affecting aerosol delivery and to correlate in vitro predictions of aerosol delivery with in vivo end points independent of patient response. Test aerosols of albuterol and antibiotics were compared. Bench measurements of inhaled mass (percentage of nebulizer charge, mean +/- SEM) ranged from 5.7 +/- 0.5% to 37.4 +/- 1.6%, with breath-actuated nebulization and humidity identified as the most important factors determining aerosol delivery. In patients, sputum levels of deposited antibiotics varied from 1.10 to 19.6 microg/ml/mg. Variation in sputum levels correlated with predictions from the in vitro model. Aerosol delivery in ventilated patients can be efficient and reproducible only if defined ventilator parameters are tightly controlled. Key parameters can be determined via in vitro bench testing defining delivery standards for clinical trials of drugs with narrow therapeutic/toxicity ratios. PMID- 12893645 TI - Are rhinovirus-induced airway responses in asthma aggravated by chronic allergen exposure? AB - Airway inflammation in asthma may represent a favorable environment for respiratory viral infections, augmenting virus-induced exacerbations in asthma. We postulated that repeated low-dose allergen exposure preceding experimental rhinovirus 16 (RV16) infection increases the severity of RV-induced airway obstruction and inflammation. Thirty-six house dust mite-allergic patients with mild to moderate asthma participated in a three-arm, parallel, placebo controlled, double-blind study. Patients inhaled a low dose of house dust mite allergen for 10 subsequent working days (Days 1-5 and 8-12) and/or were subsequently infected with RV16 (Days 15 and 16). Allergen exposure resulted in a significant fall in FEV1 (p < 0.001) and provocative concentration of histamine causing a 20% fall in FEV1 (p < 0.001) and an increase in exhaled nitric oxide (p < 0.001) and percentage of sputum eosinophils (p < 0.001). RV16 infection led to a fall in FEV1 (p = 0.02) and increases in the percentage of sputum neutrophils (p = 0.01), sputum interleukin-8 (p = 0.04), and neutrophil elastase (p = 0.04). Successive allergen exposure and RV16 infection had no synergistic or additive effect on any of the clinical or inflammatory outcomes. In conclusion, repeated low-dose allergen exposure and RV16 infection induce distinct inflammatory profiles within the airways in asthma without apparent interaction between these two environmental triggers. This suggests that preceding allergen exposure, at the used dose and duration, is not a determinant of the severity of RV-induced exacerbations in patients with mild to moderate asthma. PMID- 12893646 TI - Lung deposition and systemic availability of fluticasone Diskus and budesonide Turbuhaler in children. AB - Pharmacokinetic studies can be used to measure lung dose of inhaled drugs. The aim of this study was to compare the lung deposition of budesonide (BUD) inhaled from Turbuhaler (AstraZeneca, Lund, Sweden) and fluticasone propionate (FP) inhaled from Diskus (GlaxoSmithKline, London, UK) and to assess if the study design used for pharmacokinetic studies can be simplified. Plasma levels of BUD and FP were measured for 21 hours on five separate days in 15 patients aged 8 to 14 years: (1) Intravenous infusion of 200 microg BUD, (2) intravenous infusion of 200 microg fluticasone dipropionate, (3) inhalation of 800 microg BUD via Turbuhaler, (4) inhalation of 750 microg FP via Diskus, and (5) inhalation of BUD and FP on the same day. Charcoal was ingested to eliminate drug uptake from the gastrointestinal tract. The mean lung deposition of drug after Turbuhaler and Diskus inhalation was 30.8 and 8.0% when BUD and fluticasone were administered on separate days and 29.5% (BUD) and 7.6% (fluticasone) when the two drugs were inhaled on the same day. Lung deposition is four times higher in children after inhalation from Turbuhaler than after inhalation from Diskus. Pharmacokinetic studies with BUD and FP can be simplified because the two treatments can be administered on the same day. PMID- 12893647 TI - Cost-effectiveness of ultrasound in preventing femoral venous catheter-associated pulmonary embolism. AB - Femoral central venous catheter use is complicated by a high risk of deep venous thrombosis despite antithrombotic prophylaxis. Although some have recommended screening for femoral catheter-associated thrombosis to prevent pulmonary embolism (PE), this strategy's economic implications are unclear. Therefore, we used a decision model to evaluate the potential cost-effectiveness of a Doppler ultrasound-based screening strategy versus no ultrasound in averting thromboembolic complications associated with femoral catheters. The base-case analysis included a hypothetical cohort of 60-year-old medical patients treated for acute respiratory failure. The perspective was that of the health care payor, and the primary outcomes were quality-adjusted life expectancy, PE, and PE associated deaths. The ultrasound strategy cost $8,688/quality-adjusted life-year (QALY) gained, $5,305/PE averted, and $99,286/PE death averted. The best- and worst-case scenarios, calculated in multiway sensitivity analyses by varying in hospital mortality, deep venous thrombosis prevalence, and ultrasound accuracy, ranged from $1,170/QALY to $35,342/QALY, respectively. Probablistic analyses, in which variables with uncertain values were varied randomly within their ranges, demonstrated median costs of $12,793/QALY (interquartile range $8,176/QALY, $20,648/QALY). In summary, ultrasound screening may improve outcomes among the critically ill with femoral venous catheters at acceptable costs and could complement venous thrombosis primary prevention programs. PMID- 12893648 TI - Prospective study of air pollution and bronchitic symptoms in children with asthma. AB - The relationship of bronchitic symptoms to ambient particulate matter and to particulate elemental and organic carbon (OC), nitrogen dioxide (NO2), and other gaseous pollutants was examined in a cohort of children with asthma in 12 Southern California communities. Symptoms, assessed yearly by questionnaire from 1996 to 1999, were associated with the yearly variability of particulate matter with aerodynamic diameter less than 2.5 microg (odds ratio [OR] 1.09/microg/m3; 95% confidence interval [CI] 1.01-1.17), OC (OR 1.41/microg/m3; 95% CI 1.12 1.78), NO2 (OR 1.07/ppb; 95% CI 1.02-1.13), and ozone (OR 1.06/ppb; 95% CI 1.00 1.12). The ORs associated with yearly within-community variability in air pollution were larger than the effect of the between-community 4-year average concentrations. In two pollutant models, the effects of yearly variation in OC and NO2 were only modestly reduced by adjusting for other pollutants, except in a model containing both OC and NO2; the effects of all other pollutants were reduced after adjusting for OC or NO2. We conclude that OC and NO2 deserve greater attention as potential causes of the chronic symptoms of bronchitis in children with asthma and that previous cross-sectional studies may have underestimated the risks associated with air pollution. PMID- 12893649 TI - Cigarette smoking impairs the therapeutic response to oral corticosteroids in chronic asthma. AB - The study was designed to assess the effect of cigarette smoking on the therapeutic response to oral corticosteroids in chronic stable asthma. We performed a randomized, placebo-controlled, crossover study with prednisolone (40 mg daily) or placebo for 2 weeks in smokers with asthma, ex-smokers with asthma, and never-smokers with asthma. All subjects had reversibility in FEV1 after nebulized albuterol of 15% or more and a mean postbronchodilator FEV1% predicted of more than 80%. Efficacy was assessed using FEV1, daily PEF, and an asthma control score. There was a significant improvement after oral prednisolone compared with placebo in FEV1, ml (mean difference, 237; 95% confidence intervals, 43, 231; p = 0.019), morning PEF L/m (mean difference, 36.8; 95% confidence intervals (CI), 11, 62; p = 0.006), and asthma control score (mean difference, -0.72; 95% CI, -1.2, -0.3; p = 0.004) in never-smokers with asthma but no change in smokers with asthma (mean differences of 47, 6.5, and -0.05 with p values of 0.605, 0.47, and 0.865, respectively). Ex-smokers with asthma had a significant improvement in morning and night PEF (mean difference, 29.1; CI, 2.3, 56; p = 0.04 and 52.4; CI, 26, 79; p = 0.003, respectively), but not in FEV1 or asthma control score. We conclude that active smoking impairs the efficacy of short-term oral corticosteroid treatment in chronic asthma. PMID- 12893650 TI - Airway inflammation in childhood asthma. AB - Airway pathology has been extensively investigated in adulthood asthma, whereas only few studies examined bronchial biopsies in childhood asthma. To evaluate the airway pathology in children with asthma, we analyzed bronchial biopsies obtained from 23 children undergoing bronchoscopy for clinical indications other than asthma. Nine had mild/moderate asthma. Six had atopy without asthma, and eight had no atopy or asthma. We measured basement membrane thickness and quantified the number of eosinophils, mast cells, neutrophils, macrophages, T lymphocytes, and positive cells for transforming growth factor-beta1 (TGF-beta1) and its receptors I and II (TGFbeta-RI and TGFbeta-RII) in subepithelium. Children with asthma had an increase in basement membrane thickness and in the number of eosinophils compared with control subjects, but not compared with children with atopy. They also had a decreased expression of TGFbeta-RII compared with both those with atopy and control subjects. In children with asthma, the number of eosinophils correlated negatively with TGFbeta-RII and positively with symptom duration. In conclusion, airway eosinophilia and basement membrane thickening, which are the pathologic features that are characteristic of adulthood asthma, are already present in children with mild asthma, and even in children with atopy without asthma. Moreover, in children with asthma but not in children with atopy without asthma, there is a downregulation of TGFbeta-RII. PMID- 12893651 TI - Intrinsic cardiac nervous system in tachycardia induced heart failure. AB - The purpose of this study was to test the hypothesis that early-stage heart failure differentially affects the intrinsic cardiac nervous system's capacity to regulate cardiac function. After 2 wk of rapid ventricular pacing in nine anesthetized canines, cardiac and right atrial neuronal function were evaluated in situ in response to enhanced cardiac sensory inputs, stimulation of extracardiac autonomic efferent neuronal inputs, and close coronary arterial administration of neurochemicals that included nicotine. Right atrial neuronal intracellular electrophysiological properties were then evaluated in vitro in response to synaptic activation and nicotine. Intrinsic cardiac nicotine sensitive, neuronally induced cardiac responses were also evaluated in eight sham operated, unpaced animals. Two weeks of rapid ventricular pacing reduced the cardiac index by 54%. Intrinsic cardiac neurons of paced hearts maintained their cardiac mechano- and chemosensory transduction properties in vivo. They also responded normally to sympathetic and parasympathetic preganglionic efferent neuronal inputs, as well as to locally administered alpha-or beta-adrenergic agonists or angiotensin II. The dose of nicotine needed to modify intrinsic cardiac neurons was 50 times greater in failure compared with normal preparations. That dose failed to alter monitored cardiovascular indexes in failing preparations. Phasic and accommodating neurons identified in vitro displayed altered intracellular membrane properties compared with control, including decreased membrane resistance, indicative of reduced excitability. Early-stage heart failure differentially affects the intrinsic cardiac nervous system's capacity to regulate cardiodynamics. While maintaining its capacity to transduce cardiac mechano- and chemosensory inputs, as well as inputs from extracardiac autonomic efferent neurons, intrinsic cardiac nicotine-sensitive, local-circuit neurons differentially remodel such that their capacity to influence cardiodynamics becomes obtunded. PMID- 12893652 TI - Brain regions expressing Fos during thermoregulatory behavior in rats. AB - We surveyed the neural substrata for behavioral thermoregulation with immunohistochemical analysis of the expression of Fos protein in the rat brain. We used an operant system in which a rat exposed to heat (40 degrees C) could get cold air (0 degrees C) for 30 s when it moved into the reward area. Rats moved in and out of the reward area of the system periodically and thus maintained their body temperature at a normal level. In the rats performing heat escape behavior (active group), strong Fos immunoreactivity (Fos-IR) was found in the median preoptic nucleus (MnPO), parastrial nucleus (PS), and dorsomedial hypothalamus (DMH) compared with the controls. Another group of rats (passive group) were given the same temperature changes, regardless of the rat's movement, as those obtained by rats of the active group. Fos-IR in the MnPO was also seen in this group. The present results suggest that the PS and DMH play an important role in the genesis of thermoregulatory behavior, whereas the MnPO may be important for detecting changes in ambient and/or body temperatures. PMID- 12893653 TI - Thermoregulatory and metabolic changes during fever in young and old rats. AB - We injected old and young rats with lipopolysaccharide (LPS; 50 microg/kg ip) at two ambient temperatures (Ta; 21 and 31 degrees C). Young rats mounted equivalent fevers at both Tas [peak body temperatures (Tb) of 38.3 and 38.7 degrees C, respectively]. The Tb of old rats was not different from baseline (37.3 degrees C) after LPS at Ta 21 degrees C, whereas, at 31 degrees C, their Tb rose to a mean peak of 38.4 degrees C. We also measured the associated thermoregulatory responses by use of calorimetry. At 21 degrees C, young rats developed a fever by increasing both O2 consumption and heat conservation. Old rats did not become febrile, and O2 consumption fell by 15%. Heat loss was the same in old and young rats. At 31 degrees C, young and old rats developed similar fevers with similar increases in heat production and conservation. Our results suggest that the lack of LPS fever in old rats at 21 degrees C is due mainly to the lowered metabolic rate. PMID- 12893654 TI - Pressor effect of electroacupuncture on hemorrhagic hypotension. AB - Neiguan (PC-6) is a traditional acupoint in each forearm and overlies the trunk of the median nerve. Previous studies show that electroacupuncture (EA) at the Neiguan acupoint could improve not only myocardial ischemic dysfunction by inducing a depressor response but also recover hemorrhagic hypotension by inducing a pressor response. However, their physiological mechanisms are not yet elucidated. We investigated the pressor effect of Neiguan EA and its mechanism by focusing on left ventricular (LV) performance in a canine hemorrhagic hypotension model. We hemorrhaged 36 anesthetized and thoracotomized mongrel dogs and decreased LV end-systolic pressure (ESP) to approximately 70 mmHg (35% decrease). We obtained LV pressure-volume (P-V) data with a micromanometer catheter and a conductance catheter. One-hour Neiguan EA significantly recovered the decreased ESP, end-diastolic volume, and stroke volume by 32 +/- 13%, 27 +/- 13%, and 39 +/ 17%, respectively (P < 0.05), without changing heart rate and the slope of the end-systolic P-V relation. Neiguan EA inhibited a hemorrhage-induced increase in plasma catecholamines. However, vecuronium (neuromuscular blocking agent) administration abolished the antihypotension effect of Neiguan EA. Furthermore, Neiguan EA was much more effective than a nonacupoint thigh EA. We conclude that Neiguan EA achieved the antihypotension effect by improving LV filling of the hemorrhage-depressed LV performance despite the inhibition of the hemorrhage increased plasma catecholamines. This pressor effect seemed to accompany an increased venous return by Neiguan EA-increased vasomotor tone and muscle pump. This study demonstrated a scientific basis for the therapeutic efficacy of acupuncture in the treatment of hemorrhagic hypotension and shock. PMID- 12893655 TI - RhoA-Rho kinase mediates synergistic ET-1 and phenylephrine contraction of rat corpus cavernosum. AB - Maintenance of the detumescent state of the penis is believed to involve the actions of several vasoconstrictors. However, our mechanistic understanding of any synergistic vasoconstrictor influences is extremely limited. We tested the hypothesis that a vasoconstrictor combination of endothelin (ET-1) and phenylephrine (PE) augments the constrictor responses in rat corporal cavernosal tissues by a mechanism involving the RhoA-Rho kinase pathway. Independently, ET-1 (1 nM-30 microM) and PE (100 nM-100 microM) both caused dose-dependent contractions of isolated rat cavernosal tissues. In combination, ET-1 (30 nM) augmented the contractile effect of PE and shifted the calculated EC50 for PE (90 +/- 12 to 45 +/- 5 microM). The active stress generated by cavernosal strips during the ET-1 + PE combined stimulation (4.9 +/- 0.2 mN/mm2) was greater than the combined stress generated with ET-1 (0.4 +/- 0.1 mN/mm2) or PE (3.3 +/- 0.2 mN/mm2) stimulations alone. Blockade of ETA receptors (30 nM; A-127722) reversed the augmented stress generation and the Rho-kinase inhibitor Y-27632 differentially and dose-dependently relaxed the tissue. The combined constrictor effect was associated with a fourfold increase of RhoA in the membrane faction of the tissue homogenates. We conclude that the ET-1 + PE combination potentiate vasoconstriction through mutual activation of the RhoA-Rho kinase pathway. The interactions of these agonists likely play important roles in the maintenance of the flaccid state and contribute to some forms of erectile dysfunction. PMID- 12893656 TI - Effects of cortisol and prolactin on Na+ and Cl- transport in cultured branchial epithelia from FW rainbow trout. AB - The electrophysiological and ion-transporting properties of cultured gill epithelia from freshwater (FW) rainbow trout were examined in the presence of cortisol and prolactin as media supplements. Epithelia were of the double-seeded insert (DSI) type containing both pavement cells (PVCs) and mitochondria-rich cells (MRCs) and were grown in Leibovitz's L15 media on filters allowing exposure to different apical media conditions. Experiments were carried out in two series after 7-9 days symmetrical (L15 apical-L15 basolateral) culture. In both series, 100% L15 was maintained as the basolateral medium throughout and supplemented with physiologically relevant doses of either prolactin (50 ng/ml), cortisol (500 ng/ml), or cortisol + prolactin (500 + 50 ng/ml, respectively). In series 1, epithelia were exposed to progressively diluted apical media (100, 75, 50, 25, 12.5% L15, and FW) at 24-h intervals. The preparations retained integrity [high transepithelial resistance (TER); low ion efflux rates] during this prolonged dilution protocol. Cortisol, or cortisol + prolactin, resulted in a greater TER and reduced ion efflux rates during dilution, likely an effect on junctional permeability of PVCs, but did not promote active Na+ and Cl- uptake from apical FW. In series 2, epithelia were directly exposed to apical FW and ion fluxes measured over the first 6 h. Under these conditions, cortisol or cortisol + prolactin promoted active uptake of both Na+ and Cl- simultaneously from apical FW, probably attributable to actions on the MRCs. However, Na+-K+-ATPase activities were not significantly altered by any of the treatments in either series. Overall, prolactin alone did not appear to promote FW adaptation but exhibited synergism with cortisol. These results provide further support for the cultured DSI epithelium as an in vitro model for ion transport in FW fish. PMID- 12893657 TI - User and carer involvement in mental health services: from rhetoric to science. PMID- 12893658 TI - Global burden of depression: the intersection of culture and medicine. PMID- 12893659 TI - Protecting altruism: a call for a code of ethics in British psychiatry. PMID- 12893660 TI - Cognitive-behavioural therapy for schizophrenia: filling the therapeutic vacuum. PMID- 12893661 TI - Suicide in ethnic minority groups. PMID- 12893662 TI - Clinical trials of antidepressant medications are producing meaningless results. PMID- 12893663 TI - Evidence-based mental health policy: a critical appraisal. AB - BACKGROUND: Arguments for and against evidence-based psychiatry have mostly centred on its value for clinical practice and teaching. Now, however, use of the same paradigm in evaluating health care has generated new problems. AIMS: To outline the development of evidence-based health care; to summarise the main critiques of this approach; to review the evidence now being employed to evaluate mental health care; and to consider how the evidence base might be improved. METHOD: The following sources were monitored: publications on evidence-based psychiatry and health care since 1990; reports of randomised trials and meta analytic reviews to the end of 2002; and official British publications on mental health policy. RESULTS: Although evidence-based health care is now being promulgated as a rational basis for mental health planning in Britain, its contributions to service evaluation have been distinctly modest. Only 10% of clinical trials and meta-analyses have been focused on effectiveness of services, and many reviews proved inconclusive. CONCLUSIONS: The current evidence-based approach is overly reliant on meta-analytic reviews, and is more applicable to specific treatments than to the care agencies that control their delivery. A much broader evidence base is called for, extending to studies in primary health care and the evaluation of preventive techniques. PMID- 12893664 TI - Brain volume, asymmetry and intellectual impairment in relation to sex in early onset schizophrenia. AB - BACKGROUND: Accumulating evidence suggests that early-onset schizophrenia arises from a disturbance in the normal trajectory of cerebral development. AIMS: To investigate brain structure, asymmetry and IQ in early-onset schizophrenia. METHOD: Volumes of left and right cerebral hemispheres and IQ were assessed in 33 participants with early-onset DSM-IV schizophrenia and 30 members of a matched, normal control group. RESULTS: Total brain volume was significantly smaller in the group with early-onset disease ('cases') relative to the control group (4.5%), especially for the left hemisphere in males (6.0%). A significant sex x diagnosis interaction in hemisphere asymmetry revealed that the female cases group had significantly reduced rightward asymmetry relative to the female control group and that the male cases tended to have reduced leftward asymmetry relative to the male control group. Decreased left hemisphere volume in males and decreased rightward hemispheric asymmetry in females correlated with reduced IQ. CONCLUSIONS: Sexually dimorphic alterations in asymmetry correlate with degree of intellectual impairment in early-onset schizophrenia. PMID- 12893665 TI - Depression status, medical comorbidity and resource costs. Evidence from an international study of major depression in primary care (LIDO). AB - BACKGROUND: Despite the burden of depression, there remain few data on its economic consequences in an international context. AIMS: To explore the relationship between depression status (with and without medical comorbidity), work loss and health care costs, using cross-sectional data from a multi-national study of depression in primary care. METHOD: Primary care attendees were screened for depression. Those meeting eligibility criteria were categorised according to DSM-IV criteria for major depressive disorder and comorbid status. Unit costs were attached to self-reported days absent from work and uptake of health care services. RESULTS: Medical comorbidity was associated with a 17-46% increase in health care costs in five of the six sites, but a clear positive association between costs and clinical depression status was identified in only one site. CONCLUSIONS: The economic consequences of depression are influenced to a greater (and considerable) extent by the presence of medical comorbidity than by symptom severity alone. PMID- 12893666 TI - Assertive outreach teams in London: models of operation. Pan-London Assertive Outreach Study, part 1. AB - BACKGROUND: Assertive outreach teams have been introduced in the UK, based on the assertive community treatment (ACT) model. It is unclear how models of community care translate from one culture to another or the degree of adaptation that may result. AIMS: To characterise London assertive outreach teams and determine whether there are distinct groups within them. METHOD: Semi-structured interviews with team managers plus one month's prospective process of care data collection were used to test for 'model fidelity' to ACT and, by cluster analysis, to identify groupings. RESULTS: Fidelity varied widely, with four teams (out of 24 studied) rated 'high fidelity' and three teams rated 'low fidelity' by US standards and 17 rated 'ACT-like'. Three clusters were identified, with voluntary sector teams being the most distinct group. CONCLUSIONS: There is wide variation in the practice of assertive outreach in London. The role of the voluntary sector requires increased attention. Heterogeneity in practice is a clinical challenge but a research opportunity in distinguishing effective from redundant components of the approach. PMID- 12893667 TI - Assertive outreach teams in London: staff experiences and perceptions. Pan-London Assertive Outreach Study, part 2. AB - BACKGROUND: The job satisfaction, burn-out and work experiences of assertive outreach team staff are likely to be important to the model's sustainability. AIMS: To describe self-reported views and work experiences of staff in London's 24 assertive outreach teams and to compare these with staff in community mental health teams (CMHTs) and between different types of assertive outreach team. METHOD: Confidential staff questionnaires in London's assertive outreach teams (n=187, response rate=89%) and nine randomly selected CMHTs (n=114, response rate=75%). RESULTS: Staff in assertive outreach teams and CMHTs were moderately satisfied with their jobs, with similar sources of satisfaction and stress. Mean scores were low or average for all sub-scales of the Maslach Burnout Inventory for the assertive outreach team and the CMHT staff, with some differences suggesting less burn-out in the assertive outreach teams. Nine of the 24 assertive outreach teams had team means in the high range for emotional exhaustion and there were significant differences between types of assertive outreach team in some components of burn-out and satisfaction. CONCLUSIONS: These findings are encouraging, but repeated investigation is needed when assertive outreach teams have been established for longer. PMID- 12893668 TI - Assertive outreach teams in London: patient characteristics and outcomes. Pan London Assertive Outreach Study, part 3. AB - BACKGROUND: Although the model of assertive outreach has been widely adopted, it is unclear who receives assertive outreach in practice and what outcomes can be expected under routine conditions. AIMS: To assess patient characteristics and outcome in routine assertive outreach services in the UK. METHOD: Patients (n=580) were sampled from 24 assertive outreach teams in London. Outcomes--days spent in hospital and compulsory hospitalisation--were assessed over a 9-month follow-up. RESULTS: The 6-month prevalence rate of substance misuse was 29%, and 35% of patients had been physically violent in the past 2 years. During follow up, 39% were hospitalised and 25% compulsorily admitted. Outcome varied significantly between team types. These differences did not hold true when baseline differences in patient characteristics were controlled for. CONCLUSIONS: Routine assertive outreach serves a wide range of patients with significant rates of substance misuse and violent behaviour. Over a 9-month period an average of 25% of assertive outreach patients can be expected to be hospitalised compulsorily. Differences in outcome between team types can be explained by differences in patient characteristics. PMID- 12893669 TI - Suicides in ethnic minorities within 12 months of contact with mental health services. National clinical survey. AB - BACKGROUND: Information on suicide by psychiatric patients from ethnic minority groups is scarce. AIMS: To establish the number of patients from ethnic minorities who kill themselves; to describe their suicide methods, and their social and clinical characteristics. METHOD: A national clinical survey was based on a 4-year sample of suicides in England and Wales. Detailed data were collected on those who had been in contact with mental health services in the year before death. RESULTS: In total 282 patients from ethnic minorities died by suicide--6% of all patient suicides. The most common method of suicide was hanging; violent methods were more common than in White patient suicides. Schizophrenia was the most common diagnosis. Ethnic minority patients were more likely to have been unemployed than White patients and to have had a history of violence and recent non-compliance. In around half, this was the first episode of self-harm. Black Caribbean patients had the highest rates of schizophrenia (74%), unemployment, living alone, previous violence and drug misuse. CONCLUSIONS: In order to reduce the number of suicides by ethnic minority patients, services should address the complex health and social needs of people with severe mental illness. PMID- 12893670 TI - Clozapine v. conventional antipsychotic drugs for treatment-resistant schizophrenia: a re-examination. AB - BACKGROUND: Although there is a consensus that clozapine is more effective than conventional antipsychotic drugs for treatment-resistant schizophrenia, there is great heterogeneity among results of relevant trials. AIMS: To re-evaluate the evidence comparing clozapine with conventional antipsychotics and to investigate sources of heterogeneity. METHOD: Individual studies were inspected with assessment of clinical relevance of results. Meta-regression analysis was performed to investigate sources of heterogeneity. RESULTS: Ten trials were examined. Recent large-scale studies have not found a substantial advantage for clozapine, especially in terms of a clinically relevant effect. Meta-regression showed that shorter study duration, financial support from a drug company and higher baseline symptom score consistently predicted greater advantage of clozapine. CONCLUSIONS: It may be inappropriate to combine studies in meta analysis, given the degree of heterogeneity between their findings. The benefits of clozapine compared with conventional treatment may not be substantial. PMID- 12893671 TI - Integrating evaluative research and community-based mental health care in Verona, Italy. PMID- 12893672 TI - Explanatory models in psychiatry. PMID- 12893673 TI - Ethnic differences in prisoners: describing trauma and stress. PMID- 12893674 TI - Specialist care for prisoners? PMID- 12893675 TI - Consent and treatment in prisons. PMID- 12893676 TI - Amisulpride-induced mania in a patient with schizophrenia. PMID- 12893677 TI - Changing use of ECT. PMID- 12893678 TI - Stigma as a cause of suicide. PMID- 12893679 TI - Inappropriate use of psychostimulants. PMID- 12893680 TI - Psychiatry in pictures. PMID- 12893681 TI - Social capital and mental health v. objective measures of health in The Netherlands. PMID- 12893682 TI - Extracellular superoxide dismutase polymorphism in mice. AB - OBJECTIVE: In this study, we describe a previously unrecognized murine extracellular superoxide dismutase (ecSOD) allele and examine its distribution among various strains and its effect on the ecSOD phenotype. METHODS AND RESULTS: Polymerase chain reaction analysis of genomic and cDNA from apolipoprotein E/LDLR /- mice indicates the presence of 2 distinct transcripts for this enzyme independent of the extent of atherosclerosis or age. Sequencing and genotyping analyses reveal the presence of 2 alleles for ecSOD. One is a short variant with a 10-base pair deletion in the 3'UTR, accompanied by a single nucleotide substitution (position 61) found in the 129P3/J strain of mice. By contrast, all other strains examined carry the long form. Both free and heparin-releasable ecSOD activities in the 129P3/J strain are more than 3-fold higher than those in the C57Bl/6 mice. Corresponding differences in plasma enzyme mass are observed by immunoblotting. A clear allele dose effect can be observed in F2 hybrids of these 2 strains; free and total ecSOD activities in mice homozygous for the short allele are twice those of mice homozygous for the long allele, with the heterozygote values in between. CONCLUSIONS: These data clearly demonstrate the allele-specific effects on the ecSOD phenotype independent of other strain specific factors and underline the need for backcrossing of genetically modified mice. PMID- 12893683 TI - Ginkgo biloba extract inhibits tumor necrosis factor-alpha-induced reactive oxygen species generation, transcription factor activation, and cell adhesion molecule expression in human aortic endothelial cells. AB - OBJECTIVE: This study was conducted to examination whether Ginkgo biloba extract (GBE), a Chinese herb with antioxidant activity, could reduce cytokine-induced monocyte/human aortic endothelial cell (HAEC) interaction, a pivotal early event in atherogenesis. METHODS AND RESULTS: Pretreatment of HAECs with GBE (50 and 100 microg/mL for 18 hours) significantly suppressed cellular binding between the human monocytic cell line U937 and tumor necrosis factor-alpha (TNF-alpha) stimulated HAECs by using in vitro binding assay (68.7% and 60.1% inhibitions, respectively). Cell enzyme-linked immunosorbent assay and immunoblot analysis showed that GBE (50 microg/mL for 18 hours) significantly attenuated TNF-alpha induced cell surface and total protein expression of vascular cellular adhesion molecule-1 and intracellular adhesion molecule-1 (63.5% and 69.2%, respectively; P<0.05). However, pretreatment with probucol (5 micromol/L for 18 hours) reduced the expression of vascular cellular adhesion molecule-1 but not intracellular adhesion molecule-1. Preincubation of HAECs with GBE or probucol significantly reduced intracellular reactive oxygen species formation induced by TNF-alpha (76.8% and 68.2% inhibitions, respectively; P<0.05). Electrophoretic mobility shift assay demonstrated that both GBE and probucol inhibited transcription factor nuclear factor-kappaB activation in TNF-alpha-stimulated HAECs (55.2% and 65.6% inhibitions, respectively) but only GBE could inhibit the TNF-alpha stimulated activator protein 1 activation (45.1% inhibition, P<0.05). CONCLUSIONS: GBE could reduce cytokine-stimulated endothelial adhesiveness by downregulating intracellular reactive oxygen species formation, nuclear factor kappaB and activator protein 1 activation, and adhesion molecule expression in HAECs, supporting the notion that the natural compound Ginkgo biloba may have potential implications in clinical atherosclerosis disease. PMID- 12893684 TI - Crucial role of type 2 inositol 1,4,5-trisphosphate receptors for acetylcholine induced Ca2+ oscillations in vascular myocytes. AB - OBJECTIVE: The aim of this study was to correlate the expression of InsP3R subtypes in native vascular and visceral myocytes with specific Ca2+-signaling patterns. METHODS AND RESULTS: By Western blot and immunostaining, we showed that rat portal vein expressed InsP3R1 and InsP3R2 but not InsP3R3, whereas rat ureter expressed InsP3R1 and InsP3R3 but not InsP3R2. Acetylcholine induced single Ca2+ responses in all ureteric myocytes but only in 50% of vascular myocytes. In the remaining vascular myocytes, the first transient peak was followed by Ca2+ oscillations. By correlating Ca2+ signals and immunostaining, we revealed that oscillating vascular cells expressed both InsP3R1 and InsP3R2 whereas nonoscillating vascular cells expressed only InsP3R1. Acetylcholine-induced oscillations were not affected by inhibitors of ryanodine receptors, Ca2+ ATPases, Ca2+ influx, and mitochondrial Ca2+ uniporter but were inhibited by intracellular infusion of heparin. Using specific antibodies against InsP3R subtypes, we showed that acetylcholine-induced Ca2+ oscillations were specifically blocked by the anti-InsP3R antibody. These data were supported by antisense oligonucleotides targeting InsP3R2, which selectively inhibited Ca2+ oscillations. CONCLUSIONS: Our results suggest that in native smooth muscle cells, a differential expression of InsP3R subtypes encodes specific InsP3 mediated Ca2+ responses and that the presence of the InsP3R2 subtype is required for acetylcholine-induced Ca2+ oscillations in vascular myocytes. PMID- 12893685 TI - Ets motifs are necessary for endothelial cell-specific expression of a 723-bp Tie 2 promoter/enhancer in Hprt targeted transgenic mice. AB - OBJECTIVE: Tie-2 is an endothelial cell-specific receptor tyrosine kinase that is involved in the remodeling of blood vessels and angiogenesis. Our goal was to characterize Tie-2 promoter function as a means of providing insight into the mechanisms of endothelial cell-specific gene regulation. METHODS AND RESULTS: When targeted to the Hprt locus of mice, a small Tie-2 promoter fragment (containing a 300-bp intronic enhancer coupled upstream to a 423-bp core promoter) (T-short) directed widespread endothelial cell expression in vivo. The T-short promoter contains 2 clusters of Ets sites, one in the first exon, the other in the intronic enhancer. In cultured endothelial cells, a combined mutation of the Ets motifs resulted in a significant reduction in promoter activity. Consistent with these results, the same Ets mutations resulted in a loss of detectable expression of the T-short promoter in all vascular beds with the notable exception of the brain. CONCLUSIONS: These results suggest that the T short promoter contains information for widespread expression in the vascular tree, Ets sites are necessary for in vivo promoter activity, and the shorter Tie 2 fragment may be useful as a tool to direct heterologous gene expression within the intact endothelium. PMID- 12893686 TI - Absence of p53 leads to accelerated neointimal hyperplasia after vascular injury. AB - OBJECTIVE: It has been suggested that deregulated expression of the tumor suppressor protein p53 may play a role in the pathogenesis of occlusive vascular remodeling. However, the role of p53 in cell proliferation and apoptosis in vascular lesions has been controversial. METHODS AND RESULTS: We tested the potential involvement of p53-mediated molecular signaling in lesion formation using a mouse model of vascular injury that may resemble balloon angioplasty. A large wire was inserted into the femoral artery of p53+/+ and p53-/- mice. There was no significant difference in the occurrence of rapid-onset apoptosis, that is, 4 hours after injury. At 2 weeks, the number of proliferating cells in the lesion of p53-/- mice was significantly higher than that observed in p53+/+ mice. The frequency of apoptotic cells was significantly lower in p53-/- mice than in p53+/+ mice. At 4 weeks, the neointimal hyperplasia of p53-/- mice was greater than that of p53+/+ mice. There was no significant difference in the frequency of apoptosis in the lesions. CONCLUSIONS: These results indicate a crucial role of p53 in pathological vascular remodeling after mechanical injury and provide the basis for the development of new therapies targeting p53 for a prophylactic treatment of vascular diseases. PMID- 12893688 TI - Serum chitotriosidase activity is increased in subjects with atherosclerosis disease. AB - OBJECTIVE: This study was undertaken to analyze the relation between serum activity of chitotriosidase enzyme, a protein synthesized exclusively by activated macrophages, and atherosclerotic lesion extent in subjects with atherothrombotic stroke (ATS) and in subjects with ischemic heart disease (IHD). METHODS AND RESULTS: We assayed the serum chitotriosidase activity and a common chitotriosidase gene polymorphism that causes deficiency in chitotriosidase activity in 3 Spanish populations, ATS (n=153), IHD (n=124), and control (n=148) subjects. Statistical differences were found in serum chitotriosidase activity between ATS (88.1+/-4.6 nmol/mL. h, P<0.0001) and IHD subjects (79.0+/-6.3, P=0.002) versus control group (70.9+/-5.2). These observed differences were not attributable to a distinct allelic or genotype distribution. The extension of the atherosclerotic lesion in carotids of ATS subjects was measured by duplex sonography. Chitotriosidase activities were 66.9+/-9.6, 88.7+/-8.3, and 107.7+/ 11.8 for subjects with carotid stenosis 60%, respectively. Statistical differences were observed between subjects with major and intermediate stenosis grade compared with subjects with minor stenosis, P=0.005 and P=0.016, respectively. CONCLUSIONS: Serum chitotriosidase activity is significantly increased in individuals suffering from atherosclerosis disease and is related to the severity of the atherosclerotic lesion, suggesting a possible role as atherosclerotic extent marker. PMID- 12893687 TI - Effects of intravenous apolipoprotein A-I/phosphatidylcholine discs on LCAT, PLTP, and CETP in plasma and peripheral lymph in humans. AB - OBJECTIVE: We have previously shown that intravenous apolipoprotein A I/phosphatidylcholine (apoA-I/PC) discs increase plasma pre-beta HDL concentration and stimulate reverse cholesterol transport (RCT) in humans. We have now investigated the associated changes in the following 3 HDL components that play key roles in RCT: lecithin:cholesterol acyltransferase (LCAT), cholesteryl ester transfer protein (CETP), and phospholipid transfer protein (PLTP). METHODS AND RESULTS: apoA-I/PC discs (40 mg/kg over 4 hours) were infused into 8 healthy men. Samples of blood and prenodal peripheral lymph were collected for 24 to 48 hours. At 12 hours, plasma LCAT concentration had increased by 0.40+/-0.90 mg/L (+7.8%; mean+/-SD; P<0.05), plasma cholesterol esterification rate by 29.0+/-9.0 nmol/mL per h (+69.5%; P<0.01), plasma CETP concentration by 0.5+/-0.2 mg/L (+29.7%; P<0.01), and plasma PLTP activity by 1.45+/-0.67 micromol/mL per h (+23.9%; P<0.01). In contrast, plasma PLTP concentration had decreased by 4.4+/-2.7 mg/L (-44.8%; P<0.01). The changes in PLTP were accompanied by alterations in the relative proportions of large lipoproteins containing inactive PLTP and small particles containing PLTP of high specific activity. No changes were detected in peripheral lymph. CONCLUSIONS: Nascent HDL secretion may induce changes in PLTP, LCAT, and CETP that promote RCT by catalyzing pre-beta HDL production, cholesterol esterification in HDLs, and cholesteryl ester transfer from HDLs to other lipoproteins. PMID- 12893689 TI - Mechanisms involved in adenosine triphosphate--induced platelet aggregation in whole blood. AB - OBJECTIVE: Effects on platelet aggregation of adenosine triphosphate (ATP) released from damaged cells and from platelets undergoing exocytosis have not been clearly established. In this study we report on the effects of ATP on platelet aggregation in whole blood. METHODS AND RESULTS: Aggregation, measured using a platelet-counting technique, occurred in response to ATP and was maximal at 10 to 100 micromol/L. It was abolished by MRS2179, AR-C69931, and creatine phosphate/creatine phosphokinase, implying that conversion to adenosine diphosphate (ADP) is required. ATP did not induce aggregation in platelet-rich plasma, but aggregation did occur when apyrase or hexokinase was added. Aggregation also occurred after addition of leukocytes to platelet-rich plasma (as a source of ecto-ATPase), and this was potentiated on removal of adenosine by adenosine deaminase, indicating that adenosine production modulates the response. Dipyridamole, which inhibits adenosine uptake into erythrocytes, inhibited aggregation induced by ATP in whole blood, and adenosine deaminase reversed this. DN9693 and forskolin synergized with dipyridamole to inhibit ATP-induced aggregation. CONCLUSIONS: ATP induces aggregation in whole blood via conversion of ATP to ADP by ecto-ATPases on leukocytes. This is inhibited by agents that prevent adenosine removal. Reduced aggregation at high concentrations of ATP (>100 micromol/L) may be a consequence of inhibition by ATP of ADP action at ADP receptors. PMID- 12893690 TI - Hypercholesterolemia blunts forearm vasorelaxation and enhances the pressor response during acute systemic hypoxia. AB - OBJECTIVE: During hypoxia, active substances released by the endothelium play a key role in the cardiovascular and respiratory responses elicited to optimize oxygen delivery. As hypercholesterolemia is a major cause of endothelial dysfunction, it may interfere with these responses. METHODS AND RESULTS: We studied cardiovascular and ventilatory responses to acute systemic hypoxia in 14 patients with hypercholesterolemia (HC) and 13 control (CO) subjects. Oxygen saturation decreased similarly in both groups. Diastolic blood pressure increased only in the HC group (P=0.0002) and, despite systolic blood pressure increases both in the HC group, 140+/-4 (95% confidence interval [CI],131 to 149 mm Hg) to 154+/-4 mm Hg (95% CI,145 to 164 mm Hg), and in the CO group, 133+/-3 (95% CI,126 to 140 mm Hg) to 140+/-4 mm Hg (95% CI,132 to 148 mm Hg), the HC group showed an enhanced pressor response (P=0.03, group comparison). Both groups had increased forearm blood flow, but the decrease in forearm vascular resistance in the CO group, 40+/-5 (95% CI, 30 to 51 UR) to 31+/-4 UR (95% CI,23 to 39 UR) (P=0.0001) was not seen in the HC group, 29+/-3 (95% CI, 22 to 37 UR) to 26+/-3 UR (95% CI, 20 to 33 UR), (P=0.03, group comparison). CONCLUSIONS: Hypercholesterolemic patients demonstrate a hyperreactive pressor response and absence of forearm vasodilation during acute systemic hypoxia. PMID- 12893691 TI - Contrast media and the kidney: European Society of Urogenital Radiology (ESUR) guidelines. AB - The Contrast Media Safety Committee of the European Society of Urogenital Radiology (ESUR) has looked at the effects of contrast media on the kidney including prevention of contrast medium induced nephropathy. This has resulted in four reports dealing with 1) contrast medium induced nephrotoxicity, 2) haemodialysis and contrast media, 3) use of gadolinium contrast media instead of iodinated contrast media and 4) contrast media injection in diabetic patients receiving metformin. The review presents an overview of these four reports and offers the current understanding of the interaction between contrast agents and the kidney. PMID- 12893692 TI - Picture archiving and communication systems: the users' view. AB - A study was undertaken to assess the acceptability to users of a Picture Archiving and Communication System (PACS). A questionnaire was issued to 208 users. Some questions required a graduated score in response and others a free text response. 101 responses were received from a cross section of hospital staff. The majority of users judged PACS to be a major advance for the hospital, with high quality images, less frustration than using film and an improvement in their working lives. They judged that it had made a positive change in their working practices and had met their expectations. They welcomed the access it gave them to Radiology reports. There was more ambivalence about the impact it had made to the efficiency of clinic consultations and the conduct of ward rounds. Free text responses were made concerning the benefits and disadvantages of PACS. Analysis of these demonstrated many more benefits than disadvantages. The Radiology department has been enabled to address the concerns of staff raised through the questionnaire, and has been encouraged by the positive responses. PACS has been accepted well by a wide cross section of hospital staff. It has improved their working lives and made a major contribution to the working of the hospital as a whole. PMID- 12893693 TI - Evaluation of magnetic resonance myelography in the investigation of cervical spondylotic radiculopathy. AB - The objective of this study was to prospectively evaluate the accuracy of MR myelography for the demonstration of foraminal nerve root impingement in cervical spondylotic radiculopathy. 40 patients with cervical spondylotic radiculopathy were imaged with conventional MRI and with MR myelography. The diagnostic accuracy of these imaging strategies for the demonstration of exit foraminal stenosis was calculated relative to a gold standard of the combination of conventional MRI and MR myelography. Conventional MRI had a sensitivity of 88.9%, specificity of 99.1%, and diagnostic accuracy of 94.5% for the demonstration of exit foraminal disease (p<0.001). MR myelography alone had a sensitivity of 84.4%, a specificity of 90.1%, and diagnostic accuracy of 88% (p<0.001). However, the addition of MR myelography increased the diagnostic yield of the MR examination for the detection of foraminal stenotic disease. MR myelography is a useful adjunct to conventional MRI in the investigation of cervical spondylotic radiculopathy. PMID- 12893694 TI - Red flags in patients presenting with headache: clinical indications for neuroimaging. AB - Headache is a very common patient complaint but secondary causes for headache are unusual. Neuroimaging is both expensive and has a low yield in this group. Most patients with intracranial pathology have clinical features that would raise a "red flag". Appropriate selection of patients with headache for neuroimaging to look for secondary causes is very important. Red flags act as screening tools to help in identifying those patients presenting with headache who would benefit from prompt neuroimaging, and may increase the yield. The aim of this study is to evaluate clinical features in patients with headache using neuroimaging as a screening tool for intracranial pathology. 20 red flags were defined. A retrospective study of 111 patients was performed and the outcomes were divided into positive and negative. Abnormal neuroimaging was present in 39 patients. Results were analysed using the Logistic Regression model. Sensitivity and specificity of red flags were analysed to establish the cut-off point to predict abnormal neuroimaging and a receiver operating characteristic (ROC) curve plotted to show the sensitivity of the diagnostic test. Three red flag features proved to be statistically significant with the p-value of less than 0.05 on both univariate and multivariate analysis. These were: paralysis; papilloedema; and "drowsiness, confusion, memory impairment and loss of consciousness". In addition, if three or more red flags from the list were present, this showed strong indication of abnormal neuroimaging, from cut-off point of ROC curve (area under the curve =0.76). PMID- 12893695 TI - Multislice helical CT: the value of multiplanar image reconstruction in assessment of the bronchi and small airways disease. AB - We examined 23 consecutive patients (11 males and 12 females with mean age of 56 years) with possible airway diseases to assess the impact of multiplanar image reconstruction (MPR) on the degree of confidence and accuracy in diagnosing bronchial abnormalities and emphysema. The thorax was scanned contiguously at 1 mm slice thickness using Siemens Volume Zoom Multislice CT scanner. Images were reconstructed at 1 mm slice thickness (lung windows L-600HU W-1600HU utilizing high spatial frequency algorithm) in the axial (10 mm apart), sagittal (4 images per lung) and coronal (6 images) plane. Paddle wheel image reconstructions were also performed in the assessment of bronchiectasis. Axial images were assessed with and without the help of MPR by three chest radiologists at two separate occasions (at least 4 weeks apart). The presence of bronchiectasis, emphysema and bronchiolitis in each lobe was documented on a confidence scale of 0 to 3. The overall mean confidence for each observer with and without MPR was compared. Consensus diagnosis was used as the gold standard for the assessment of the diagnostic accuracy of each observer. A confidence score of 2 or more for any lobe was considered diagnostic of the particular airway disease. The diagnostic accuracy for each observer with and without MPR was compared. Consensus reporting diagnosed bronchiectasis in 7 patients (30.4%), bronchiolitis in 5 patients (21.7%) and emphysema in 12 patients (52%). MPR did not increase the confidence of assessing the different abnormalities for all observers but improvement in diagnosing bronchiectasis was noted in two observers. The improvement did not reach statistical significance. However, agreement between observers in the diagnosis of bronchiectasis and emphysema was improved when the MPR images were used in conjunction with standard axial imaging (Kappa statistic improved from 0.29 to 0.54 for bronchiectasis and from 0.7 to 0.81 for emphysema). Agreement on the diagnosis of bronchiolitis was not improved by MPR for all observers. Our results suggest that MPR seems to improve the confidence in diagnosing bronchiectasis and emphysema. PMID- 12893696 TI - A contribution to the establishment of diagnostic reference levels in CT. AB - CT has become the major source of population exposure to diagnostic X-rays. CT dose index (CTDI) and dose-length product (DLP) have been proposed as the appropriate dose quantities for the establishment of diagnostic reference levels for optimizing patient exposure. Dose measurements on 27 CT scanners in Northern Greece involving six routine CT examinations have been performed in order to compare their performance with the currently proposed European reference dose values and to produce a preliminary set of data for the establishment of local diagnostic reference levels. All measurements were performed using a pencil shaped ionization chamber introduced into polymethyl methacrylate cylindrical head and body phantoms. The results revealed significant discrepancies in dose values among the CT scanners, which can be mainly attributed to variations in the examination protocols and the different kinds of scanners. Significant overdosing compared with the European reference levels has not been observed, with the exception of the routine head examination, where 47% of the scanners exceeded the corresponding CTDI(w) value. CT scans in the trunk region result in the higher effective doses, which can reach estimated maximal values of the order of 15 mSv. PMID- 12893697 TI - A study on radiation doses and irradiated areas in cerebral embolisation. AB - Patient radiation doses during interventional radiology procedures may reach the thresholds for radiation-induced skin and eye lens injuries. This study investigates the irradiated areas and doses received by patients undergoing cerebral embolisation, which is regarded as a high dose interventional radiology procedure. For each procedure the fluoroscopic and digital dose-area product (DAP), the fluoroscopic time, the total number of acquired images and entrance skin dose (ESD) calculated by the angiographic unit were recorded. The ESD was measured by means of thermoluminescent dosimeters. In this study, the skin, eye and thyroid gland doses and the irradiated area for 30 patients were recorded. The average ESD was found to be 0.77 Gy for the posteroanterior plane and 0.78 Gy for the lateral plane. The average DAP was 48 Gy cm(2) for the posteroanterior plane and 58 Gy cm(2) for the lateral plane. The patient's average right eye dose was 60 mGy and the dose to the thyroid gland was 24 mGy. Seven patients received a dose above 1 Gy, one patient exceeded the threshold for transient erythema and one exceeded the threshold for temporary epilation. A good correlation between the DAP and the ESD for both planes has been found. The doctor's eye dose has also been measured for 17 procedures and the average dose per procedure was 0.13 mGy. PMID- 12893698 TI - Comparison between human pharmacokinetics and imaging properties of two conjugation methods for 99mTc-annexin A5. AB - Annexin A5 (AnxA5) is a protein with high affinity for phosphatidyl serine, a phospholipid exposed on the cell surface during apoptosis. This phenomenon has been used for determination of cell death after myocardial infarction. To evaluate the potential of (99m)Tc-AnxA5 for in vivo scintigraphy of apoptotic cells, the pharmacokinetics and imaging properties of two radiopharmaceuticals, (99m)Tc-(n-1-imino-4-mercaptobutyl)-AnxA5 (I-AnxA5) and (99m)Tc-(4,5 bis(thioacetamido)pentanoyl)-AnxA5 (B-AnxA5), were studied. I-AnxA5 was administered intravenously to seven patients and one healthy volunteer, and B AnxA5 was administered to 12 patients. All patients in the pharmacokinetic study had myocardial disease. Additionally, imaging was performed in a patient with acute myocardial infarction, as well as in three patients with different malignancies. The plasma concentration, excretion and biodistribution of (99m)Tc AnxA5 were measured, as well as levels of AnxA5 antigen. The kinetic data of both radiopharmaceuticals in plasma fitted a two-compartment model. Both preparations had similar half-lives, but a different distribution over the two compartments. Plasma levels of AnxA5 antigen showed a broad variation. Both radiopharmaceuticals accumulated in the kidney, liver and gut. B-AnxA5 was excreted significantly faster than I-AnxA5. Both compounds can be used for imaging of the head/neck region, the thorax and the extremities. B-AnxA5 has a faster clearance and a lower radiation dose. Imaging of apoptosis in the abdomen will be difficult with both radiopharmaceuticals, and especially with B-AnxA5 because of its faster appearance in the gut. PMID- 12893699 TI - Breast density: agreement of measures from film and digital image. AB - Mammographic density, in particular density from digital images, is increasingly used in breast cancer research. We investigated the concordance between density assigned by the same radiologist to a mammogram film and a digital image of the same mammogram. Two density measures were investigated, Wolfe parenchymal patterns and a six category classification (SCC) system of density. Included in the study were 78 women, 528 mammograms. Crude and weighted Kappa statistics were used to estimate agreement between the density assigned from the film and the image. Kappa for Wolfe measures was 71%, p<0.001 and for SCC measures was 54%, p<0.001. Weighted Kappa values were 79%, p<0.001 and 77%, p<0.001, respectively. There was some evidence to suggest that the digitized image may be assigned a higher Wolfe but not numerical category than the original film, and the magnitude of these differences was small. Neither age nor mammogram view (craniocaudal or mediolateral oblique) were related to the likelihood of agreement of the two density measurements. This evidence justifies the use of digital images in the visual assessment of breast density in research studies. PMID- 12893700 TI - Ureteric obstruction and intravascular administration of contrast media: is there a risk? PMID- 12893701 TI - MRI diagnosis of bilateral adrenal vein thrombosis. AB - We report a case of bilateral adrenal vein thrombosis in an adult female who had a history of breast cancer. The patient does not have clinical, serological or imaging evidence of metastatic disease 14 months from the initial diagnosis. Adrenal vein thrombosis is a rare entity. There have been no previous reports specifically stating an association between adrenal vein thrombosis and hypercoaguability, but there are many cases in the literature documenting venous thrombosis elsewhere within the body in patients with hypercoaguable states. Laboratory testing performed to exclude a hypercoaguable state, revealed heterozygosity for the Factor V Leiden mutation/activated protein C resistance and elevated factor VIII levels [3660 IU l(-1) (<1500)]. This is the first reported case of bilateral metachronous adrenal vein thrombosis in which MRI established the diagnosis. PMID- 12893702 TI - Idiopathic presacral retroperitoneal fibrosis: report of two cases. AB - Retroperitoneal fibrosis is characteristic plaques of dense fibrous tissue, typically extending from the level of the kidney down to the sacral promontory. Involvement of the presacral space is extremely rare in retroperitoneal fibrosis. We report two cases of idiopathic retroperitoneal fibrosis manifested by a presacral mass. PMID- 12893703 TI - Pedunculated malignant peripheral nerve-sheath tumour of the diaphragm presenting as recurrent lung infection: ultrasound diagnosis. AB - We report a rare case of malignant peripheral nerve-sheath tumour of the diaphragm in a 12-year-old boy with neurofibromatosis presented clinically and radiographically as recurrent lung infection. Ultrasound revealed a pedunculated diaphragmatic tumour invading the posterior chest wall. This case highlights that this unusual tumour may limit diaphragmatic excursion leading to, and even being masked by, superimposed lung infection. In patients with neurofibromatosis, ultrasound is recommended when chest radiography shows a basal lung opacity. PMID- 12893704 TI - A case of unilateral enophthalmos. PMID- 12893707 TI - The inflammatory response: friend or enemy for muscle injury? AB - Limiting certain aspects of inflammation may be a useful new treatment for sport related muscle injury. PMID- 12893708 TI - Exercise related transient abdominal pain. AB - The causes of exercise related transient abdominal pain remain to be elucidated. PMID- 12893709 TI - Abnormal biomechanics: a precursor or result of knee osteoarthritis? AB - Biomedical studies are required to differentiate between the causes and results of knee osteoarthritis. PMID- 12893710 TI - Rehabilitation of ligamentous ankle injuries: a review of recent studies. AB - There are many treatment modalities for ankle rehabilitation. These are reviewed, and the most effective training programme for rapid restoration of ankle movement, strength, endurance, and proprioception is selected. PMID- 12893711 TI - Effects of supramaximal exercise on the electromyographic signal. AB - AIM: To determine the neuromuscular recruitment characteristics during supramaximal exercise. METHODS: Ten healthy subjects completed the Wingate anaerobic test (WAT) cycling protocol. Electromyographic (EMG) data and rate of fatigue were recorded throughout the cycling. RESULTS: The mean (SD) rate of fatigue (decrease in power output) was 44.5 (8.6)%. No significant change was found in EMG amplitude. A significant decrease (p<0.01) in mean power frequency spectrum was found over the 30 second period. CONCLUSIONS: During WAT, mean power frequency spectrum was attenuated with no decline in EMG amplitude, which may be caused by an accumulation of metabolites in the periphery. However, it is also possible that the feedback loop from intramuscular metabolism to the central nervous system is unable, within the 30 second period of the WAT, to affect neural recruitment strategy. PMID- 12893712 TI - Effects of different sodium concentrations in replacement fluids during prolonged exercise in women. AB - OBJECTIVE: To investigate the effect of different sodium concentrations in replacement fluids on haematological variables and endurance performance during prolonged exercise. METHODS: Thirteen female endurance athletes completed three four hour runs on a 400 m track. Environmental conditions differed between the three trials: 5.3 degrees C and snow (trial 1), 19.0 degrees C and sunny weather (trial 2), 13.9 degrees C and precipitation (trial 3). They consumed 1 litre of fluid an hour during the trials with randomised intake of fluids: one trial (H) with high sodium concentration (680 mg/l), one trial (L) with low sodium concentration (410 mg/l), and one trial with only water (W). Before and after the trials, subjects were weighed and blood samples were taken for analysis of [Na(+)](plasma), packed cell volume, and mean corpuscular volume. RESULTS: The mean (SD) decrease in [Na(+)](plasma) over the whole trial was significantly (p<0.001) less in trial H (2.5 (2.5) mmol/l) than in trial W (6.2 (2.1) mmol/l). Mild hyponatraemia ([Na(+)](plasma) = 130-135 mmol/l) was observed in only six women (46%) in trial H compared with nine (69%) in trial L, and 12 (92%) in trial W. Two subjects (17%) in trial W developed severe hyponatraemia ([Na(+)](plasma)<130 mmol/l). No significant differences were found in performance or haematological variables with the three different fluids. There was no significant correlation between[Na(+)](plasma) after the run and performance. There was a significant correlation between changes in [Na(+)](plasma) and changes in body weight. CONCLUSIONS: Exercise induced hyponatraemia in women is likely to develop from fluid overload during prolonged exercise. This can be minimised by the use of replacement fluids of high sodium concentration. Sodium replacement of at least 680 mg/h is recommended for women in a state of fluid overload during endurance exercise of four hours. However, higher [Na(+)](plasma) after the run and smaller decreases in [Na(+)](plasma) during the trials were no indication of better performance over four hours. PMID- 12893713 TI - Effect of exercise on upper respiratory tract infection in sedentary subjects. AB - OBJECTIVE: To determine if exercise training affects the severity and duration of a naturally acquired upper respiratory tract infection (URTI) in sedentary subjects. METHODS: Subjects were sedentary volunteers (two or fewer days a week of exercise for less than 30 minutes a day for the previous three months), 18-29 years of age, with a naturally acquired URTI (three to four days of onset). All subjects were screened-for example, asthma, hay fever-by a doctor and were afebrile. Volunteers were alternately assigned to an exercise (EX) group (four men, seven women) or a non-exercise (NEX) group (three men, eight women). Subjects in the EX group completed 30 minutes of supervised exercise at 70% of target heart rate range for five days of a seven day period. For the initial screening, and every 12 hours, all subjects completed a 13 item symptom severity checklist and a physical activity log. Cold symptom scores were obtained until the subjects were asymptomatic. Significance was set at p/==" BORDER="0">50 ml/kg/min. Four measures of athletic performance were tested: VO(2)MAX, anaerobic capacity (anaerobic speed test), aerobic endurance (time to fatigue at 90% of VO(2)MAX), and isokinetic strength (Cybex II dynamometer). Height, weight, and six skinfold measurements were also recorded. All these observational tests were completed during both the follicular and mid-luteal phases of an ovulatory menstrual cycle. Cycle phases were confirmed by assaying plasma oestradiol and progesterone. Participants were subsequently randomly assigned to either a tricyclic oral contraceptive or placebo and retested in identical fashion (oral contraceptive phase). RESULTS: Absolute and relative changes in VO(2)MAX from follicular to oral contraceptive phase decreased in the oral contraceptive group by 4.7%, whereas the placebo group showed a slight increase (+1.5%) over the same time period. Two of the women taking oral contraceptive had decreases of 4 and 9 ml/kg/min. In contrast, most women in the placebo group improved or maintained VO(2)MAX. There was also a significant increase in the sum of skinfolds in women taking oral contraceptive compared with those taking placebo (p<0.01). There were no significant changes in other physiological variables (maximum ventilation, heart rate, respiratory exchange ratio, packed cell volume) or measures of performance (anaerobic speed test, aerobic endurance, isokinetic strength) as a function of oral contraceptive treatment. CONCLUSIONS: The decrease in VO(2)MAX that occurs when oral contraceptive is taken may influence elite sporting performance in some women. Further studies are required to determine the mechanisms of this change. PMID- 12893717 TI - A 16 year study of injuries to professional boxers in the state of Victoria, Australia. AB - OBJECTIVES: Concerns about the significant injury risks in boxers have been well documented. To inform the continuing debate, updated information about the risk of injury for participants, and suitable means of modifying or preventing these risks, need to be identified. METHODS: Data describing all professional boxing fight outcomes and injuries sustained during competition, from August 1985 to August 2001, were obtained from the Victorian Professional Boxing and Combat Sports Board. RESULTS: A total of 107 injuries were recorded from 427 fight participations, corresponding to an injury rate of 250.6 injuries per 1000 fight participations. The most commonly injured body region was the head/neck/face (89.8%), followed by the upper extremities (7.4%). Specifically, injuries to the eye region (45.8%) and concussion (15.9%) were the most common. About three quarters of all injuries were lacerations/open wounds or superficial. No information was available on the mechanism of injury. CONCLUSIONS: Future research should collect information on the mechanism of injury, as this is crucial for the development and implementation of effective injury prevention strategies. A suggested boxing injury report form is provided to facilitate this. PMID- 12893718 TI - Evaluation of cricket helmet performance and comparison with baseball and ice hockey helmets. AB - BACKGROUND: Protective helmets in sport are important for reducing the risk of head and facial injury. In cricket and other sports with projectiles, national test standards control the minimum helmet performance. However, there are few field data showing if helmets are effective in reducing head injury. OBJECTIVES: (a) To examine the performance of cricket helmets in laboratory tests; (b) to examine performance with regard to test standards, game hazards, and helmet construction; (c) to compare and contrast these findings with baseball and ice hockey helmets. METHODS: Impact tests were conducted on a selection of helmet models: five cricket, two baseball, and two ice hockey. Ball to helmet impacts at speeds of 19, 27, 36, and 45 m/s were produced using an air cannon and a Hybrid III dummy headform and neck unit. Free fall drop tests with a rigid headform on to a selection of anvils (flat rigid, flat deformable, and hemispherical rigid) were conducted. Resultant headform acceleration was measured and compared between tests. RESULTS: At the lower speed impacts, all helmets produced a good reduction in headform acceleration, and thus injury risk. At the higher speed impacts, the effectiveness was less. For example, the mean maximum headform accelerations for all cricket helmets at each speed were: 67, 160, 316, and 438 g for 19, 27, 36, and 45 m/s ball speeds respectively. Drop tests on to a hemispherical anvil produced the highest accelerations. The variation in performance increased as the magnitude of the impact energy increased, in both types of testing. CONCLUSIONS: The test method used for baseball helmets in which the projectile is fired at the helmet may be superior to helmet drop tests. Cricket helmet performance is satisfactory for low speed impacts, but not for impacts at higher, more realistic, speeds. Baseball and ice hockey helmets offer slightly better relative and absolute performance at the 27 m/s ball and puck impacts. PMID- 12893719 TI - Reliability of stabilised commercial dynamometers for measuring hip abduction strength: a pilot study. AB - BACKGROUND: Reliable quantification of hip abductor strength in a clinical setting is challenging. OBJECTIVES: To examine the intrarater and interrater reliability of three commonly used commercial dynamometers in the measurement of hip abduction. METHODS: Supine gravity minimised measures of unilateral hip abduction strength were recorded in 10 women (mean (SD) age 23.5 (1.9) years) using three different commercially available dynameters. Measurements were repeated over a three day period with a different device used on each day. RESULTS: Intrarater reliability ranged from 0.880 to 0.958 across the three devices, and measures of interrater reliability ranged from 0.899 to 0.948. CONCLUSION: Commercially available dynamometers can be used to quantify hip abduction strength with good to excellent reliability. A previously undescribed method of quantifying hip abduction strength in a clinical setting using readily available instrumentation is presented. PMID- 12893720 TI - General practitioners and doping in sport: attitudes and experience. AB - OBJECTIVES: To examine the attitudes to, and knowledge of, doping in sport of French general practitioners (GPs), and their contact with drug taking athletes on an everyday basis. METHODS: A total of 402 GPs were randomly selected from all over France and interviewed by telephone, using a prepared script. RESULTS: The response rate was 50.5% (153 men and 49 women; mean (SD) age 45.6 (5.6) years). Of the respondents, 73% confirmed that they had the list of banned products, and only 34.5% stated that they were aware of the latest French law, brought into effect in March 1999, concerning the fight against doping. Some 11% had directly encountered a request for prescription of doping agents over the preceding 12 months (the requested substances were mainly anabolic steroids, stimulants, and corticosteroids), and 10% had been consulted by an athlete who was using doping drugs and was frightened of the health risks (the substances used were mainly anabolic steroids). Over half (52%) of the GPs favoured the prescription of drug substitutions to athletes who used doping agents. According to 87.5% of respondents, doping is a public health problem, and 80% stated that doping is a form of drug addiction. Most (89%) said that a GP has a role to play in doping prevention, but 77% considered themselves poorly prepared to participate in its prevention. CONCLUSION: The results suggest that (a) GPs have limited knowledge of doping and (b) are confronted with doping in their daily practice, at least occasionally. PMID- 12893721 TI - Kinematic and electromyographic analysis of the push movement in tai chi. AB - BACKGROUND: Tai chi is a form of exercise derived from the martial art folk traditions of China. The force used in tai chi includes different principles of mechanical advantage. No studies on the kinematic features of tai chi exercise have been published. OBJECTIVE: To analyse the kinematics and electromyographic characteristics of tai chi. METHODS: An experienced tai chi master was asked to perform a sequence of basic movements: ward off, roll back, press, and push. The movements were videotaped and digitised using a motion analysis system. Electromyographic activities of the lumbar erector spinae, rectus femoris, medial hamstrings, and medial head of gastrocnemius were recorded by surface electrodes. The push movement data were analysed. RESULTS: The medial hamstrings and medial head of gastrocnemius muscle groups maintained low activity, with higher electromyographic values in the lumbar erector spinae and substantially higher ones in the rectus femoris during the push movement. Both concentric and eccentric contractions occurred in muscles of the lower limbs, with eccentric contraction occurring mainly in the anti-gravity muscles such as the rectus femoris and the medial head of gastrocnemius. The forward and backward shifts in centre of gravity (CG) were mainly accomplished by increasing and decreasing respectively the joint angles of the bilateral lower limbs rather than by adopting a forward or backward postural lean. The path of the CG in the anteroposterior and mediolateral component was unique, and the sway or deviation from the path was small. The master maintained an upright posture and maintained a low CG (hips, knees, and ankles bent) while travelling slowly and steadily from one position to another. CONCLUSION: The eccentric muscle contraction of the lower limbs in the push movement of tai chi may help to strengthen the muscles. PMID- 12893723 TI - Don't save the ball! AB - OBJECTIVES: To identify, over a period of 12 months, all attendances at an accident and emergency department by children over the age of 5 years with an injury to a wrist, hand, or finger, and to examine those sustained as a result of a blow from a ball. METHODS: The case notes of all children aged 6-13 years attending the Accident and Emergency Department of the Royal Aberdeen Children's Hospital in the year 2001 as a result of a wrist, hand, or finger injury sustained from a blow by a ball were reviewed, and the cause, type, and severity of the injury noted. RESULTS: A total of 187 children were identified; 69% were boys and football was the main sport involved. Most (93%) were radiographed, and 40% were positive; most fractures were sustained outwith school. Hand dominance was not significant. CONCLUSIONS: These injuries are common and may be preventable with modification of rules, equipment, and coaching. A register of youth sporting injuries may show if there are any detrimental long term effects. PMID- 12893722 TI - Effect of testosterone and endurance training on glycogen metabolism in skeletal muscle of chronic hyperglycaemic female rats. AB - OBJECTIVES: To investigate in glycolytic and oxidative muscles of trained (nine weeks) and untrained hyperglycaemic female rats the effect of hyperandrogenicity and/or endurance training on energy metabolic properties. METHODS: Glycogen content and activity of muscle enzymes with regulatory functions in glycogen synthesis were examined. RESULTS: Testosterone treatment increased glycogen content of extensor digitorum longus (EDL) and soleus muscles of hyperglycaemic sedentary (18% and 84% respectively) and hyperglycaemic trained (7% and 16% respectively) rats. In both types of muscle of the hyperglycaemic testosterone treated exercised subgroup, less depletion of glycogen was found than in the untreated group (38% and 87% for EDL and soleus respectively). CONCLUSIONS: The mechanisms by which training and/or hyperandrogenism alone or in combination elicits their specific effects are complex. Differences in sex, surgery, levels of hormones administered, and exercise model used may be the main reasons for the observed discrepancies. Conclusions from the results: (a) hyperandrogenism is not a primary cause of the development of insulin resistance; (b) glycogen content of slow and fast twitch muscle is increased by training through increased glycogen synthase activity. The most plausible explanation for differences between different muscle fibre types is the different levels of expression of androgen receptors in these fibres. Hyperandrogenicity therefore acts on energy metabolic variables of hyperglycaemic animals by different mechanisms in glycolytic and oxidative muscle fibres. PMID- 12893724 TI - Impact of school sports injury. AB - BACKGROUND: Most injuries in school occur during sport. OBJECTIVES: To explore the impact of sports injury in supervised school sport. METHOD: A prospective study of sports injury in children of secondary school age presenting to the accident and emergency department. Each patient was identified on registration, matched with medical records after discharge, and contacted later by telephone to complete a structured interview. Patients were only included if their injury was sustained during supervised school sport. RESULTS: During the study period, 194 patients aged 11-18 attended the accident and emergency department with an injury, 51% of which occurred during school sport. Injuries occurred most commonly in rugby (43%), followed by physical education and games together (17.5%). Most injuries were x rayed (72%). Just over 12% of pupils lost no time from sport, most (71%) were back to sport within three weeks, and 2.7% were injured for more than eight weeks. Almost a third of parents needed to take time off from work to deal with the injured child. CONCLUSION: School sports injuries are important. They account for just over half of all injuries in secondary school children. They cause significant disruption to school and sport and have important implications for the wider family. PMID- 12893725 TI - Insulin as a drug of abuse in body building. AB - A theoretical benefit of insulin abuse by body builders is that it is undetectable by currently available tests. A case is presented that highlights the dangers of such abuse. PMID- 12893726 TI - Popliteus tendon rupture: a case report and review of the literature. AB - Isolated rupture of the popliteus musculotendinous unit is an uncommon injury. A case is here reported of a semiprofessional athlete with a ruptured popliteus tendon without significant instability of the knee. The diagnosis was made by magnetic resonance imaging and confirmed by arthroscopy. PMID- 12893727 TI - Avulsion fracture of the anterior superior iliac spine following apophysitis. AB - The case of an avulsion fracture occurring in an 18 year old athlete after repeated running is presented. The fracture involved the tensor fascia lata muscle and the external oblique abdominal muscle. It was treated by open reduction and internal fixation with two screws. The patient returned to full training, including long distance running, only four weeks after the operation. PMID- 12893728 TI - Unusual limb injury associated with sport parachuting. AB - The case is reported of an open injury of the right arm sustained during a parachute jump. The fracture was managed in the usual way with wound debridement, wound irrigation, skeletal stabilisation, and reconstruction of the soft tissues. Good shoulder and elbow function were achieved. The combined effort of an orthopaedic trauma surgeon and plastic surgeon is essential to improve outcomes in such cases. PMID- 12893729 TI - Isolated dislocation of the proximal tibiofibular joint in a long jumper. AB - Acute traumatic proximal tibiofibular joint dislocation is an exceedingly rare injury. This is a case report in a rare horizontal type joint variant, following a long jump injury. The diagnostic approach when this injury is suspected is described. PMID- 12893730 TI - Localised pigmented villonodular synovitis: an uncommon cause of knee pain mimicking a meniscal tear. AB - A previously well 30 year old man presented with a short history of intra articular mechanical locking, swelling, and anteromedial joint line pain. There was localised tenderness, and McMurray's test was positive. Arthroscopy revealed a 3.6 x 2.6 x 1.5 cm firm pedicular localised pigmented villonodular synovitis originating from the insertion of the anterior horn of the medial meniscus. Owing to its size and consistency, mini-arthrotomy was required. This allowed a return to sporting activities. Localised pigmented villonodular synovitis can mimic symptoms of a meniscal tear. PMID- 12893731 TI - Are Reliable Change (RC) calculations appropriate for determining the extent of cognitive change in concussed athletes? PMID- 12893732 TI - Significant association between fluctuations in serum urate and high density lipoprotein cholesterol during exhaustive training. PMID- 12893733 TI - Applying elite research to the general population. PMID- 12893734 TI - Response to "Berger in retrospect: effect of varied weight training programmes on strength". PMID- 12893735 TI - Accessory nerve injury during amateur wrestling: silent but not overlooked. PMID- 12893736 TI - Will the new field hockey rules lead to more injuries? PMID- 12893737 TI - Specific contribution of estrogen receptors on mitogen-activated protein kinase pathways and vascular cell activation. AB - Randomized clinical trials have not provided conclusive data that hormone replacement therapy confers cardioprotection against coronary artery disease in postmenopausal women. However, other studies have shown that estrogens can induce beneficial effects on the vasculature. Nevertheless, the specific contribution of estrogen receptors (ERs) alpha and beta on vascular cells is not well characterized. Therefore, we used an antisense gene therapy approach to investigate the contribution of ERalpha and ERbeta on p38 and p42/44 mitogen activated protein kinase (MAPK) activation and on vascular cell responsiveness. Treatment of porcine smooth muscle cells (PSMCs) with platelet-derived growth factor-BB induced p38 and p42/44 MAPK activation and their migration and proliferation. These effects were prevented by pretreatment with 17beta-estradiol (17betaE). The inhibitory effects of 17betaE on PSMCs were abrogated by the downregulation of ERbeta protein expression with selective ERbeta mRNA antisense oligomers, whereas the downregulation of ERalpha had no effect. However, treatment of porcine aortic endothelial cells with 17betaE promoted p38 and p42/44 MAPK phosphorylation and their migration and proliferation. These effects were ERalpha dependent, as defined by antisense gene therapy. These results suggest that in PSMCs, 17betaE reduces p42/44 and p38 MAPK activity through ERbeta stimulation, whereas in contrast, in porcine aortic endothelial cells, 17betaE induces p42/44 and p38 MAPK through ERalpha activation. 17betaE may contribute to the vascular healing process and to the prevention of restenosis by improving reendothelialization through ERalpha activation and by decreasing smooth muscle cell migration and proliferation through ERbeta stimulation. PMID- 12893738 TI - Cardiac memory is associated with decreased levels of the transcriptional factor CREB modulated by angiotensin II and calcium. AB - Cardiac memory (CM) has short- (STCM) and long-term (LTCM) components modulated by calcium and angiotensin II. LTCM is associated with reduced Ito and Kv4.3 mRNA levels. Because the cAMP response element binding protein, CREB, contributes to CNS memory transcription, we hypothesized that it might be a transcriptional factor in CM, influenced by calcium and angiotensin II. We studied STCM in dogs that were AV sequentially paced (AVP) for 2 hours or sham-operated. STCM was evaluated with ECG and vectorcardiogram (VCG), and subepicardial biopsies were taken at 5 to 120 minutes and investigated for CREB. LTCM was studied in dogs paced for 3 weeks and in sham controls. At 3 weeks the heart was excised, biopsies obtained, and CRE binding tested. STCM induction occurred in AVP dogs but not in sham or AVP dogs treated with saralasin or nifedipine. Nuclear CREB was significantly decreased at 2 hours in the AVP no-drug group only. LTCM dogs manifested reduced binding of nuclear proteins to CRE, and CRE binding activity in the promoter region of Kv4.3. In conclusion, there is an association between STCM induction and decreased nuclear CREB that is angiotensin-modulated and calcium-dependent. Moreover, the decreased CRE binding after 3 weeks of AVP combined with CRE binding activity in the Kv4.3 promoter can explain the Kv4.3 mRNA and Ito downregulation that characterize LTCM. PMID- 12893739 TI - Very-long-chain acyl-coenzyme a dehydrogenase deficiency in mice. AB - Fatty acid oxidation (FAO) defects are inborn errors of metabolism clinically associated with cardiomyopathy and sudden infant death syndrome (SIDS). FAO disorders often present in infancy with myocardial dysfunction and arrhythmias after exposure to stresses such as fasting, exercise, or intercurrent viral illness. It is uncertain whether the heart, in the absence of stress, is normal. We generated very-long-chain acyl-coenzyme A dehydrogenase (VLCAD)-deficient mice by homologous recombination to define the onset and molecular mechanism of myocardial disease. We found that VLCAD-deficient hearts have microvesicular lipid accumulation, marked mitochondrial proliferation, and demonstrated facilitated induction of polymorphic ventricular tachycardia, without antecedent stress. The expression of acyl-CoA synthase (ACS1), adipophilin, activator protein 2, cytochrome c, and the peroxisome proliferator activated receptor gamma coactivator-1 were increased immediately after birth, preceding overt histological lipidosis, whereas ACS1 expression was markedly downregulated in the adult heart. We conclude that mice with VLCAD deficiency have altered expression of a variety of genes in the fatty acid metabolic pathway from birth, reflecting metabolic feedback circuits, with progression to ultrastructural and physiological correlates of the associated human disease in the absence of stress. PMID- 12893740 TI - The obesity-associated peptide leptin induces hypertrophy in neonatal rat ventricular myocytes. AB - One of the major manifestations of obesity is increased production of the adipocyte-derived 16-kDa peptide leptin, which is also elevated in heart disease, including congestive heart failure. However, whether leptin can directly alter the cardiac phenotype is not known. We therefore studied the effect of leptin as a potential hypertrophic factor in cultured myocytes from 1- to 4-day-old neonatal rat heart ventricles. Using RT-PCR, we demonstrate that these cells express the short-form (OB-Ra) leptin receptor. Twenty-four hours of exposure to leptin (0.31 to 31.3 nmol/L) produces a significantly increased cell surface area that peaked at 0.63 nmol/L. Subsequent experiments were done with 3.1 nmol/L leptin, which significantly increased cell area by 42%, protein synthesis by 32%, and alpha-skeletal actin and myosin light chain-2 expression by 250% and 300%, respectively. These events occurred in the absence of any increased cell death. Hypertrophy was preceded by rapid activation of the mitogen-activated protein kinase system including p38 and p44/42 as early as 5 minutes after leptin addition, whereas hypertrophy was inhibited by the p38 inhibitor SB203580 but not by the p44/42 inhibitor PD98059. Our results demonstrate a direct hypertrophic effect of leptin and may offer a biological link between hypertrophy and hyperleptinemic conditions such as obesity. PMID- 12893741 TI - Ablation and mutation of nonmuscle myosin heavy chain II-B results in a defect in cardiac myocyte cytokinesis. AB - We have identified a novel form of cardiac myocyte enlargement in nonmuscle myosin heavy chain II-B (NMHC II-B) ablated mice, based on a partial failure in cytokinesis. In contrast to most cells, cardiac myocytes lack NMHC II-A, and ablation of NMHC II-B results in a heart with 70% fewer myocytes at embryonic day 14.5 (E14.5) than control mice (B+/B- and B+/B+). In addition, B-/B- cardiac myocytes show a marked increase in binucleation at E12.5, reflecting the occurrence of karyokinesis in the absence of cytokinesis. An increase in binucleation and cell size is also found in hypomorphic, homozygous mice harboring a single amino acid mutation (R709C) in the gene encoding NMHC II-B. The nonmyocytes in B-/B- hearts and homozygous mutant hearts, all of which contain NMHC II-A, do not show either of these abnormalities. B-/B- cardiac myocytes at E14.5 show a decreased bromodeoxyuridine (BrdU) labeling index compared with controls, consistent with the decrease in myocyte proliferation. This decreased BrdU labeling is not seen in nonmyocyte cells in the heart. In addition to these changes, both B-/B- mice as well as homozygous mutated mice show an increase in cyclin D2 and D3 reflecting an abnormality in earlier steps in the cell cycle. Whereas cardiac myocytes completely ablated for NMHC II-B show enlargement and binucleation, mice expressing as little as 6% of the normal amount of wild-type NMHC II-B in the heart do not show these abnormalities. PMID- 12893742 TI - Ligand-independent activation of vascular endothelial growth factor receptor 2 by fluid shear stress regulates activation of endothelial nitric oxide synthase. AB - Fluid shear stress generated by blood flowing over the endothelium is a major determinant of arterial tone, vascular remodeling, and atherogenesis. Nitric oxide (NO) produced by endothelial NO synthase (eNOS) plays an essential role in regulation of vascular function and structure by blood flow, but the molecular mechanisms that transduce mechanical force to eNOS activation are not well understood. In this study, we found that laminar flow (shear stress=12 dyne/cm2) rapidly activates vascular endothelial growth factor receptor 2 (VEGFR2) in a ligand-independent manner and leads to eNOS activation in cultured endothelial cells. Flow-stimulated VEGFR2 recruits phosphoinositide 3-kinase and mediates activation of Akt and eNOS. Inhibiting VEGFR2 kinase with selective inhibitors blocks flow-induced activation of Akt and eNOS and production of NO. Decreasing VEGFR2 expression with antisense VEGFR2 oligonucleotides significantly attenuates activation of Akt and eNOS. Furthermore, Src kinases are involved in flow stimulated VEGFR2 because inhibiting Src kinases by PP2, a selective inhibitor for Src kinases, abolishes flow-induced VEGFR2 tyrosine phosphorylation and downstream signaling. Finally, we show that inhibiting VEGFR2 kinase significantly reduces flow-mediated NO-dependent arteriolar dilation in vivo. These data identify VEGFR2 as a key mechanotransducer that activates eNOS in response to blood flow. PMID- 12893743 TI - Coupling of cardiac electrical activity over extended distances by fibroblasts of cardiac origin. AB - Roughly half of the cells of the heart consist of nonmyocardial cells, with fibroblasts representing the predominant cell type. It is well established that individual cardiomyocytes and fibroblasts in culture establish gap junctional communication at the single cell level (short-range interaction). However, it is not known whether such coupling permits activation of cardiac tissue over extended distances (long-range interaction). Long-range interactions may be responsible for electrical synchronization of donor and recipient tissue after heart transplantation and may play a role in arrhythmogenesis. This question was investigated using a novel heterocellular culture model with strands of cardiomyocytes interrupted by cardiac fibroblasts over defined distances. With use of optical recording techniques, it could be shown that impulse propagation along fibroblast inserts was successful over distances up to 300 microm and was characterized by length-dependent local propagation delays ranging from 11 to 68 ms (apparent local "conduction velocities" 4.6+/-1.8 mm/s, n=23). Involvement of mechanical stretch in this phenomenon was excluded by showing that inserts consisting of communication-deficient HeLa cells were incapable of supporting propagation. In contrast, HeLa cells expressing connexin43 permitted impulse conduction over distances as long as 600 microm. Immunocytochemistry showed that fibroblasts and cardiomyocytes expressed connexin43 and connexin45, whereas connexin40 was absent. These results illustrate that fibroblasts of cardiac origin are capable of synchronizing electrical activity of multicellular cardiac tissue over extended distances through electrotonic interactions. This synchronization is accompanied by extremely large local conduction delays, which might contribute to the generation of arrhythmias in fibrotic hearts. PMID- 12893744 TI - Fibroblast growth factor receptor-1 is essential for in vitro cardiomyocyte development. AB - Fibroblast growth factor (FGF)/FGF receptor (FGFR) signaling plays a crucial role in mesoderm formation and patterning. Heartless mutant studies in Drosophila suggest that FGFR1, among the different FGFRs, may play a role in cardiogenesis. However, fgfr1-/- mice die during gastrulation before heart formation. To establish the contribution of FGFR1 in cardiac development, we investigated the capacity of murine fgfr1+/- and fgfr1-/- embryonic stem (ES) cells to differentiate to cardiomyocytes in vitro. Clusters of pulsating cardiomyocytes were observed in >90% of 3-dimensional embryoid bodies (EBs) originated from fgfr1+/- ES cells at day 9 to 10 of differentiation. In contrast, 10% or less of fgfr1-/- EBs showed beating foci at day 16. Accordingly, fgfr1-/- EBs were characterized by impaired expression of early cardiac transcription factors Nkx2.5 and d-Hand and of late structural cardiac genes myosin heavy chain (MHC) alpha, MHC-beta, and ventricular myosin light chain. Homozygous fgfr1 mutation resulted also in alterations of the expression of mesoderm-related early genes, including nodal, BMP2, BMP4, T(bra), and sonic hedgehog. Nevertheless, fgfr1+/- and fgfr1-/- EBs similarly express cardiogenic precursor, endothelial, hematopoietic, and skeletal muscle markers, indicating that fgfr1-null mutation exerts a selective effect on cardiomyocyte development in differentiating ES cells. Accordingly, inhibitors of FGFR signaling, including the FGFR1 tyrosine kinase inhibitor SU 5402, the MEK1/2 inhibitor U0126, and the protein kinase C inhibitor GF109 all prevented cardiomyocyte differentiation in fgfr1+/- EBs without affecting the expression of the hematopoietic/endothelial marker flk-1. In conclusion, the data point to a nonredundant role for FGFR1-mediated signaling in cardiomyocyte development. PMID- 12893745 TI - Quantification of PRV-1 mRNA distinguishes polycythemia vera from secondary erythrocytosis. AB - To date, the diagnosis of polycythemia vera (PV) relies on clinical criteria. We have recently described the overexpression of a hematopoietic receptor, polycythemia rubra vera-1 (PRV-1), in patients with PV. Here, we report a quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) assay for the measurement of PRV-1 mRNA levels. We have determined PRV-1 expression in 71 patients with PV, 11 patients with secondary erythrocytosis (SE), as well as in 80 healthy controls. PV patients express significantly higher amounts of PRV-1 than healthy controls or patients with SE (P <.0001). Because there is no overlap between the PRV-1 expression in PV patients versus healthy controls or SE patients, the assay has a very high sensitivity and specificity for the diagnosis of PV in our population. In patients with erythrocytosis, the quantitative RT-PCR assay described here therefore provides a rapid, highly specific and sensitive tool for the diagnosis of PV. PMID- 12893746 TI - Cathepsin-B-dependent apoptosis triggered by antithymocyte globulins: a novel mechanism of T-cell depletion. AB - Antithymocyte globulins (ATGs), the immunoglobulin G (IgG) fraction of sera from rabbits or horses immunized with human thymocytes or T-cell lines, are used in conditioning regimens for bone marrow transplantation, in the treatment of acute graft-versus-host disease, in the prevention or treatment of acute rejection in organ transplantation, and in severe bone marrow aplasia. In nonhuman primates, ATGs induce rapid, dose-dependent, T-cell depletion in peripheral lymphoid tissues, where apoptotic cells can be demonstrated in T-cell zones. We show here that increasing ATG concentrations in vitro resulted in reduced lymphocyte proliferative responses, associated with a rapid increase in the percentage of apoptotic cells. Apoptosis did not require prior exposure to interleukin-2, nor did it result in CD178/CD95 or tumor necrosis factor/tumor necrosis factor receptor (TNF/TNF-R) interactions; it was therefore clearly different from activation-induced cell death. Cytochrome c release, caspase-9, and caspase-3 activation were not implicated, excluding a direct involvement of the intrinsic mitochondrial pathway. The cysteine protease inhibitor E64d and cathepsin-B specific inhibitors conferred significant protection, whereas apoptosis was associated with the release of active cathepsin B into the cytosol. These data demonstrate a role for cathepsin B in T-cell apoptosis induced by ATGs at concentrations achieved during clinical use. PMID- 12893747 TI - Identification and characterization of 2 types of erythroid progenitors that express GATA-1 at distinct levels. AB - Transcription factor GATA-1 is essential for the development of the erythroid lineage. To ascertain whether strict control of GATA-1 expression level is necessary for achieving proper erythropoiesis, we established transgenic mouse lines expressing green fluorescent protein (GFP) under the control of the GATA-1 gene hematopoietic regulatory domain. We examined the GATA-1 expression level by exploiting the transgenic mice and found 2 GFP-positive hematopoietic progenitor fractions in the bone marrow. One is the GFPhigh fraction containing mainly CFU-E and proerythroblasts, which coexpress transferrin receptor, while the other is the GFPlow/transferrin receptor-negative fraction containing BFU-E. Since the intensity of green fluorescence correlates well with the expression level of GATA 1, these results indicate that GATA-1 is highly expressed in erythroid colony forming unit (CFU-E) but low in erythroid burst-forming unit (BFU-E), suggesting that the incremental expression of GATA-1 is required for the formation of erythroid progenitors. We also examined GFP-positive fractions in the transgenic mouse spleen and fetal liver and identified fractions containing BFU-E and CFU-E, respectively. This study also presents an efficient method for enriching the CFU E and BFU-E from mouse hematopoietic tissues. PMID- 12893748 TI - Comparison of autologous and allogeneic hematopoietic stem cell transplantation for follicular lymphoma. AB - In this article, we report on 904 patients undergoing transplantation for follicular lymphoma. A total of 176 (19%) received allogeneic, 131 (14%) received purged autologous, and 597 (67%) received unpurged autologous transplants. Five year treatment-related mortality (TRM) rates were 30%, 14%, and 8% and 5-year recurrence rates were 21%, 43%, and 58% after allotransplantation, purged autotransplantation, and unpurged autotransplantation, respectively. In multivariate analyses, allotransplantation had higher TRM and lower disease recurrence. Purged autotransplantation had a 26% lower recurrence risk than unpurged autotransplantation. Five-year probabilities of survival were 51%, 62%, and 55% after allogeneic, purged autotransplantation, and unpurged autotransplantation, respectively. Advanced age, prolonged interval from diagnosis to transplantation, high lactate dehydrogenase (LDH), refractory disease, bone marrow involvement, low performance scores, and transplantation between 1990 and 1993 were associated with adverse outcomes. Total body irradiation was associated with higher TRM but lower recurrence. There was no association between acute or chronic graft-versus-host disease and recurrence after allotransplantation. We conclude that both allogeneic and autologous transplantation can induce durable remissions. There may be a benefit to graft purging in autologous transplantation. The decreased recurrence after allotransplantation is offset by increased TRM. We did not detect a correlation between graft-versus-host disease (GVHD) and recurrence. Finally, outcomes of transplantation for follicular lymphoma show improvement over the past decade. PMID- 12893749 TI - Functional defects of dendritic cells in patients with CD40 deficiency. AB - We have recently identified 2 patients with a rare autosomal recessive form of hyper IgM disease, known as HIGM3, caused by mutations in the CD40 gene. These patients had opportunistic infections observed on X-linked hyper IgM syndrome (HIGM), suggesting that the CD40-CD40 ligand interaction is important for promoting T-cell-mediated immunity. To evaluate whether innate immunity signals may substitute CD154 for inducing the maturation of dendritic cells (DCs), we analyzed monocyte-derived DCs in these patients. Monocyte-derived DCs of HIGM3 subjects on ex vivo stimulation with tumor necrosis factor-alpha (TNF-alpha) or lipopolysaccharide (LPS) combined with interferon-gamma (IFN-gamma) normally express all the markers of mature DCs, such as CD83 and DC-LAMP. However, cell surface levels of HLA-DR in mature DCs are reduced, as is costimulatory activity of these cells for allogeneic naive T cells. In addition, CD40-deficient DCs secrete lower amounts of interleukin-12 (IL-12) but larger quantities of IL-10 than control subjects. Finally, analysis of circulating plasmacytoid DCs demonstrates a normal percentage of this subset in CD40-deficient cells, but IFN alpha secretion in response to herpes simplex virus 1 (HSV-1) infection is severely reduced in patients. These observations suggest that the severe impairment of DC maturation may contribute to the defect of T-cell-mediated immunity observed in HIGM3 patients. PMID- 12893750 TI - Human CD8+CD25+ thymocytes share phenotypic and functional features with CD4+CD25+ regulatory thymocytes. AB - CD8+CD25+ cells, which expressed high levels of Foxp3, glucocorticoid-induced tumor necrosis factor receptor (GITR), CCR8, tumor necrosis factor receptor 2 (TNFR2), and cytotoxic T-lymphocyte-associated antigen 4 (CTLA-4) mRNAs, were identified in the fibrous septa and medullary areas of human thymus. Activated CD8+CD25+ thymocytes did not produce cytokines, but most of them expressed surface CTLA-4 and transforming growth factor beta1 (TGF-beta1). Like CD4+CD25+, CD8+CD25+ thymocytes suppressed the proliferation of autologous CD25-T cells via a contact-dependent mechanism. The suppressive activity of CD8+CD25+ thymocytes was abrogated by a mixture of anti-CTLA-4 and anti-TGF-beta1 antibodies and it was mediated by their ability to inhibit the expression of the interleukin 2 receptor alpha chain on target T cells. These results demonstrate the existence of a subset of human CD8+CD25+ thymocytes sharing phenotype, functional features, and mechanism of action with CD4+CD25+ T regulatory cells. PMID- 12893751 TI - The contributions of the alpha 2 beta 1 integrin to vascular thrombosis in vivo. AB - The alpha 2 beta 1 integrin serves as a receptor for collagens, laminin, and several other nonmatrix ligands. Many studies have suggested that the alpha 2 beta 1 integrin is a critical mediator of platelet adhesion to collagen within the vessel wall after vascular injury and that the interactions of the platelet alpha 2 beta 1 integrin with subendothelial collagen after vascular injury are required for proper hemostasis. We have used the alpha 2 beta 1 integrin deficient mouse to evaluate the contributions of the alpha 2 beta 1 integrin in 2 in vivo models of thrombosis. Studies using a model of endothelial injury to the carotid artery reveal that the alpha 2 beta 1 integrin plays a critical role in vascular thrombosis at the blood-vessel wall interface under flow conditions. In contrast, the alpha 2 beta 1 integrin is not required for the formation of thrombi and pulmonary emboli following intravascular injection of collagen. Our results are the first to document a critical in vivo role for the alpha 2 beta 1 integrin in thrombus formation at the vessel wall under conditions of shear following vascular injury. PMID- 12893752 TI - NK-cell purging of leukemia: superior antitumor effects of NK cells H2 allogeneic to the tumor and augmentation with inhibitory receptor blockade. AB - Natural killer (NK) cells are composed of subsets characterized by the expression of inhibitory or activating receptors, or both, specific for different major histocompatibility complex (MHC) class I determinants. We have previously shown that inhibitory receptor blockade of syngeneic NK cells was an effective means of ex vivo purging of leukemia-contaminated bone marrow and that the transplantation of mice with the purged bone marrow cells (BMCs) resulted in long-term, relapse free survival. We have extended the investigation to assess the antitumor effects mediated by NK cells H2-allogeneic to tumor cells. We demonstrate that various tumor cell lines are more susceptible to lysis by H2-allogeneic NK cells than by syngeneic NK cells in vitro even though comparable percentages of Ly49 NK cells were present. Using allogeneic NK cells to purge leukemia-contaminating BMCs before transplantation resulted in a higher proportion of mice with long-term survival than using syngeneic NK cells. Allogeneic NK cells did not suppress hematopoietic reconstitution as measured by granulocyte/monocyte-colony-forming unit (CFU-GM), complete blood count (CBC), and donor chimerism at various days after transplantation. Inhibitory receptor blockade of allogeneic NK cells also significantly increased these antitumor effects at lower NK/tumor ratios compared with those of syngeneic NK cells. These results demonstrate that H2-allogeneic NK cells mediate more potent antitumor effects than syngeneic NK cells without adverse hematologic effects and thus may be useful in cancer therapy. PMID- 12893753 TI - Multiple integrin-ligand interactions synergize in shear-resistant platelet adhesion at sites of arterial injury in vivo. AB - Damage to the integrity of the vessel wall results in exposure of the subendothelial extracellular matrix (ECM), which triggers integrin-dependent adhesion and aggregation of platelets. The role of platelet beta1 integrins in these processes remains mostly undefined. Here, we demonstrate by intravital fluorescence microscopy that platelet adhesion and thrombus growth on the exposed ECM of the injured carotid artery is not significantly altered in alpha2-null mice and even in mice with a Cre/loxP-mediated loss of all beta1 integrins on their platelets. In contrast, inhibition of alphaIIbbeta3 integrin on platelets in wild-type mice blocked aggregate formation and reduced platelet adhesion by 60.0%. Strikingly, alphaIIbbeta3 inhibition had a comparable effect in alpha2 null mice, demonstrating that other receptors mediate shear-resistant adhesion in the absence of functional alpha2beta1 and alphaIIbbeta3. These were identified to be alpha5beta1 and/or alpha6beta1 as alphaIIbbeta3 inhibition abrogated platelet adhesion in beta1-null mice. We conclude that shear-resistant platelet adhesion on the injured vessel wall in vivo is a highly integrated process involving multiple integrin-ligand interactions, none of which by itself is essential. PMID- 12893754 TI - CD4+CD25+ regulatory T cells inhibit immune-mediated transgene rejection. AB - Like cellular transplantation, gene therapy is often limited by immune rejection of the newly expressed antigen. In a model of gene transfer in muscle, delivery of the influenza hemagglutinin (HA) membrane protein by adeno-associated virus (AAV) is impaired by a strong immune response that leads to a rapid rejection of the transduced fibers. We show here that injection of HA-specific CD4+CD25+ T cells from T-cell receptor (TCR)-transgenic animals, concomitant with gene transfer, down-regulates the anti-HA cytotoxic and B-lymphocyte responses and enables persistent HA expression in muscle. This demonstrates for the first time that adoptive transfer of antigen-specific CD4+CD25+ regulatory T cells can be used to induce sustained transgene engraftment in solid tissues. PMID- 12893755 TI - Defective RNA ribose synthesis in fibroblasts from patients with thiamine responsive megaloblastic anemia (TRMA). AB - Fibroblasts from patients with thiamine-responsive megaloblastic anemia (TRMA) syndrome with diabetes and deafness undergo apoptotic cell death in the absence of supplemental thiamine in their cultures. The basis of megaloblastosis in these patients has not been determined. Here we use the stable [1,2-13C2]glucose isotope-based dynamic metabolic profiling technique to demonstrate that defective high-affinity thiamine transport primarily affects the synthesis of nucleic acid ribose via the nonoxidative branch of the pentose cycle. RNA ribose isolated from TRMA fibroblasts in thiamine-depleted cultures shows a time-dependent decrease in the fraction of ribose derived via transketolase, a thiamine-dependent enzyme in the pentose cycle. The fractional rate of de novo ribose synthesis from glucose is decreased several fold 2 to 4 days after removal of thiamine from the culture medium. No such metabolic changes are observed in wild-type fibroblasts or in TRMA mutant cells in thiamine-containing medium. Fluxes through glycolysis are similar in TRMA versus control fibroblasts in the pentose and TCA cycles. We conclude that reduced nucleic acid production through impaired transketolase catalysis is the underlying biochemical disturbance that likely induces cell cycle arrest or apoptosis in bone marrow cells and leads to the TRMA syndrome in patients with defective high-affinity thiamine transport. PMID- 12893756 TI - Plasticity of marrow-derived stem cells. AB - Bone marrow (BM) contains hematopoietic stem cells (HSCs), which differentiate into every type of mature blood cell; endothelial cell progenitors; and marrow stromal cells, also called mesenchymal stem cells (MSCs), which can differentiate into mature cells of multiple mesenchymal tissues including fat, bone, and cartilage. Recent findings indicate that adult BM also contains cells that can differentiate into additional mature, nonhematopoietic cells of multiple tissues including epithelial cells of the liver, kidney, lung, skin, gastrointestinal (GI) tract, and myocytes of heart and skeletal muscle. Experimental results obtained in vitro and in vivo are the subject of this review. The emphasis is on how these experiments were performed and under what conditions differentiation from bone marrow to epithelial and neural cells occurs. Questions arise regarding whether tissue injury is necessary for this differentiation and the mechanisms by which it occurs. We also consider which bone marrow subpopulations are capable of this differentiation. Only after we have a better understanding of the mechanisms involved and of the cells required for this differentiation will we be able to fully harness adult stem cell plasticity for clinical purposes. PMID- 12893757 TI - PECAM-1 negatively regulates GPIb/V/IX signaling in murine platelets. AB - Platelet adhesion at sites of vascular injury is mediated, in part, by interaction of the platelet plasma membrane glycoprotein (GP) Ib/V/IX complex with von Willebrand Factor (VWF) presented on collagen-exposed surfaces. Recent studies indicate that GPIb/V/IX may be functionally coupled with the Fc receptor gamma (FcR gamma)-chain, which, by virtue of its cytoplasmic immunoreceptor tyrosine-based activation motif, sends activation signals into the cell. Platelet endothelial cell adhesion molecule-1 (PECAM-1) is an inhibitory receptor that has previously been shown to negatively regulate platelet responses to collagen, which transduces activation signals via the GPVI/FcR gamma-chain complex. To determine whether PECAM-1 might similarly regulate signals emanating from GPIb/FcR gamma, we compared activation and aggregation responses to VWF of PECAM 1-positive and PECAM-1-deficient murine platelets. PECAM-1 and the FcR gamma chain became rapidly tyrosine phosphorylated in platelets following botrocetin induced VWF binding, but FcR gamma-chain tyrosine phosphorylation was delayed in PECAM-1-positive, versus PECAM-1-deficient, platelets. PECAM-1-deficient platelets were hyperaggregable to VWF, exhibited enhanced spreading and, under conditions of arterial flow, formed markedly larger thrombi on immobilized VWF than did wild-type platelets. Taken together, these data support the notion that engagement of the GPIb complex, in addition to sending activation signals, also initiates a negative feedback loop involving PECAM-1 that controls the rate and extent of platelet activation. PMID- 12893758 TI - Congenital afibrinogenemia: identification and expression of a missense mutation in FGB impairing fibrinogen secretion. AB - Congenital afibrinogenemia is a rare autosomal recessive disorder characterized by complete absence of detectable fibrinogen. We previously identified the first causative mutations for this disease: a homozygous deletion of approximately 11 kb of the fibrinogen alpha-chain gene (FGA). Subsequent studies revealed that the great majority of afibrinogenemia mutations are localized in FGA, but mutations were also found in FGG and FGB. Apart from 3 missense mutations identified in the C-terminal portion of FGB, all fibrinogen gene mutations responsible for afibrinogenemia are null. In this study, a young boy with afibrinogenemia was found to be a compound heterozygote for 2 mutations in FGB: an N-terminal nonsense mutation W47X (exon 2) and a missense mutation (G444S, exon 8). Coexpression of the FGB G444S mutant cDNA in combination with wild-type FGA and FGG cDNAs demonstrated that fibrinogen molecules containing the mutant beta chain are able to assemble but are not secreted into the media, confirming the pathogenic nature of the identified mutation. PMID- 12893759 TI - Clonal dominance of chronic myelogenous leukemia is associated with diminished sensitivity to the antiproliferative effects of neutrophil elastase. AB - Clinical observations suggest that in chronic myelogenous leukemia (CML), the Philadelphia chromosome (Ph+) clone has a growth advantage over normal hematopoiesis. Patients with CML have high levels of neutrophil elastase, which has recently been shown to antagonize the action of granulocyte-colony stimulating factor (G-CSF) and other growth factors. We therefore compared the effect of elastase on the growth of normal and CML progenitor cells. In 10-day suspension cultures of normal or CML CD34+ cells supplemented with G-CSF, stem cell factor (SCF), and granulocyte macrophage-colony-stimulating factor (GM-CSF), CML cells had diminished sensitivity to the growth inhibitory effect of elastase. When equal numbers of CML and normal CD34+ cells were cocultured for 10 days, there was no change in the relative proportions of normal and leukemic cells (measured by fluorescence in situ hybridization [FISH] or flow cytometry). However, when elastase was added, CML cells predominated at the end of the culture period (78% vs 22% with 1 microg/mL and 80% vs 20% with 5 microg/mL elastase). CML neutrophils substituted effectively for elastase in suppressing the proliferation of normal CD34+ cells, but this effect was abrogated by serine protease inhibitors. These results suggest that elastase overproduction by the leukemic clone can change the growth environment by digesting growth factors, thereby giving advantage to Ph+ hematopoiesis. PMID- 12893760 TI - Primary prophylaxis with warfarin prevents thrombosis in paroxysmal nocturnal hemoglobinuria (PNH). AB - Paroxysmal nocturnal hemoglobinuria (PNH) is an acquired hemolytic anemia in which venous thrombosis is the most common cause of death. Here we address the risk factors for thrombosis and the role of warfarin prophylaxis in PNH. The median follow-up of 163 PNH patients was 6 years (range, 0.2-38 years). Of the patients, 29 suffered thromboses, with a 10-year incidence of 23%. There were 9 patients who presented with thrombosis, and in the remainder the median time to thrombosis was 4.75 years (range, 3 months-15 years). The 10-year risk of thrombosis in patients with large PNH clones (PNH granulocytes > 50%) was 44% compared with 5.8% with small clones (P <.01). Patients with large PNH clones and no contraindication to anticoagulation were offered warfarin. There were no thromboses in the 39 patients who received primary prophylaxis. In comparison, 56 patients with large clones and not taking warfarin had a 10-year thrombosis rate of 36.5% (P =.01). There were 2 serious hemorrhages in more than 100 patient years of warfarin therapy. Large PNH granulocyte clones are predictive of venous thrombosis, although the exact cut-off for clone size is still to be determined. Primary prophylaxis with warfarin in PNH prevents thrombosis with acceptable risks. PMID- 12893762 TI - High-dose therapy in multiple myeloma. PMID- 12893761 TI - Transient down-modulation of CD20 by rituximab in patients with chronic lymphocytic leukemia. AB - Lymphoid cells in most patients with chronic lymphocytic leukemia (CLL), when treated with rituximab, become CD20-. This is thought to be due to masking of CD20 by rituximab. We used specific antimouse immunoglobulin antibodies to detect rituximab on the surface of CLL lymphocytes and we demonstrate that rituximab is rarely detectable after therapy. Only 3 of 65 patients with CLL had rituximab detectable on their lymphocytes after rituximab therapy despite the fact that most had no detectable CD20 expression. In vitro mixing of CLL or Raji cells with rituximab demonstrated that rituximab was detectable on the surface of cells due to its binding to CD20. However, the addition of plasma led to the down modulation of CD20 expression, and the rituximab became undetectable. This down modulation of CD20 protein expression was associated with a down-modulation of CD20 mRNA. CLL cells that lost their CD20 expression regained CD20 expression after 24 hours in culture. These data suggest that rituximab therapy leads to a substantial but transient down-modulation of CD20 expression and that negativity for CD20 in cells from patients treated with rituximab is not necessarily due to CD20 masking. The importance of this down-modulation in the efficacy of current therapy with rituximab needs further investigation. PMID- 12893763 TI - Induction of apoptosis in retinoid-refractory acute myelogenous leukemia by a novel AHPN analog. AB - Acute myelogenous leukemia (AML) is a heterogeneous disease consisting of a variety of different leukemic subtypes. While acute promyelocytic leukemia displays marked sensitivity to the differentiating effects of trans-retinoic acid (tRA), other subtypes of AML display resistance. We now describe a novel compound (E)-4-[3-(1-adamantyl)-4-hydroxyphenyl]-3-chlorocinnamic acid (3-Cl-AHPC/MM002) that induces apoptosis in the tRA-resistant leukemia cell lines M07e, KG-1, and HL-60R, and in tRA-resistant patient leukemic blasts. The 3-Cl-AHPC totally inhibits leukemia colony formation at concentrations that inhibit committed human bone marrow stem cell proliferation, that is, granulocyte/macrophage colony forming units (CFU-GMs) by only 30%. Exposure to 3-Cl-AHPC results in caspase activation and the cleavage of poly(adenosine diphosphate) (poly(ADP)) ribose polymerase. While activation of the extracellular signal-regulated kinase (ERK) and p38 pathways is not necessary for 3-Cl-AHPC-mediated apoptosis, maximal apoptosis requires c-Jun N-terminal kinase (JNK) activation. The 3-Cl-AHPC mediated cleavage of the antiapoptotic B-cell leukemia XL (Bcl-XL) protein to a proapoptotic 18-kDa product is found in both the M07e cell line and patient leukemic blasts. The 3-Cl-AHPC treatment of mice bearing the AML 1498 cell line results in a 3.3-log kill in the leukemic blasts. While 3-Cl-AHPC does not activate retinoic nuclear receptors, it is a potent inducer of apoptosis in AML cells and may represent a novel therapy in the treatment of this disease. PMID- 12893764 TI - Phenotypic correction of a mouse model of hemophilia A using AAV2 vectors encoding the heavy and light chains of FVIII. AB - Using separate adeno-associated viral 2 (AAV2) vectors to deliver the heavy and light chains of factor VIII (FVIII) we have overcome the packaging limitations of AAV, achieving phenotypic correction of hemophilia A in mice. AAV vectors were constructed that use a liver-specific promoter and the cDNA sequences of either the human or canine heavy and light chains of FVIII. After intraportal vein injection of these vectors in hemophilia-A mice, therapeutic to superphysiologic levels of active FVIII were achieved in plasma in a dose-dependent manner. Phenotypic correction of the bleeding diathesis was demonstrated by survival of all treated mice after tail clipping. Biochemical analysis demonstrated lower levels of heavy-chain (25- to 100-fold) compared with light-chain protein in the plasma of treated animals. Differences in gene transfer and transcription did not account for the differences in protein expression. We hypothesize that improvements in FVIII activity could be achieved by improvements in FVIII heavy chain expression. This work demonstrates that cotransduction of liver with AAV vectors expressing the heavy and light chains of FVIII corrects hemophilia A in vivo, providing an alternative approach to the use of a single vector. This strategy may potentially be useful for other large therapeutic proteins that contain functionally distinct domains. PMID- 12893765 TI - T-cell interaction with ICAM-1/ICAM-2 double-deficient brain endothelium in vitro: the cytoplasmic tail of endothelial ICAM-1 is necessary for transendothelial migration of T cells. AB - Endothelial intercellular adhesion molecule 1 (ICAM-1) and ICAM-2 are both involved in lymphocyte extravasation during immunosurveillance and inflammation. To define their exact role during T-cell extravasation, we used mouse T cells and ICAM-1-/-ICAM-2-/- brain endothelioma cells. ICAM-1-/-ICAM-2-/- brain endothelioma cells did not support transendothelial migration (TEM) of T cells in vitro. Re-expression of different ICAM-1 mutants in the ICAM-1-/-ICAM-2-/- endothelioma line bEndI1/2.1 or in the ICAM-1-/- endothelioma line bEndI1.1 demonstrated that the extracellular domain of ICAM-1 suffices to support T-cell adhesion while the presence of the cytoplasmic tail was strictly required for TEM. Surprisingly, tyrosine phosphorylation of endothelial ICAM-1 was not necessary for TEM of T cells or for Rho guanosine triphosphatase (RhoGTPase) activation. Furthermore, cytoplasmic deletion mutants of ICAM-1 were unable to mediate RhoGTPase activation. Thus, our data demonstrate that the cytoplasmic tail of endothelial ICAM-1-independently from tyrosine phosphorylation-is essential for supporting TEM of T lymphocytes, while Rho signaling is involved in endothelial cells. PMID- 12893766 TI - Comparative analysis of genes regulated by PML/RAR alpha and PLZF/RAR alpha in response to retinoic acid using oligonucleotide arrays. AB - Acute promyelocytic leukemia (APL) is associated with chromosomal translocations involving retinoic acid receptor alpha (RAR alpha) and its fusion partners including promyelocytic leukemia (PML) and promyelocytic leukemia zinc finger (PLZF). Using oligonucleotide arrays, we examined changes in global gene expression mediated by the ectopic expression of either PML/RAR alpha (retinoid sensitive) or PLZF/RAR alpha (retinoid-resistant) in U937 cells. Of more than 5000 genes analyzed, 16 genes were commonly up-regulated, and 57 genes were down regulated by both fusion proteins suggesting their role in the APL phenotype. In our APL model, for example, TNFAIP2, TNFR2, ELF4, RAR gamma, and HoxA1 were down regulated by both fusion proteins in the absence of retinoic acid (RA). RA strongly up-regulated these genes in PML/RAR alpha, but not in PLZF/RAR alpha expressing U937 cells. Expression studies in NB4, retinoid-resistant NB4-R2, normal human CD34+ cells, and APL patient samples strongly suggest their role in the regulation of granulocytic differentiation. Furthermore, combined treatment with tumor necrosis factor alpha (TNF alpha) and RA synergistically enhanced granulocytic differentiation in NB4 cells but not in NB4-R2 cells. Our data indicate that APL pathogenesis and retinoid-induced granulocytic differentiation of APL cells involve genes in the cell death pathway, and that cooperation between the RA and TNFalpha signaling pathways exists. Targeting both the retinoid-dependent differentiation and the cell death pathways may improve leukemic therapy, especially in retinoid-resistant acute myeloid leukemia. PMID- 12893767 TI - Physical proximity and functional interplay of PECAM-1 with the Fc receptor Fc gamma RIIa on the platelet plasma membrane. AB - We and others have recently defined that Platelet Endothelial Cell Adhesion Molecule-1 (PECAM-1/CD31) functions as a negative regulator of platelet-collagen interactions involving the glycoprotein VI/Fc receptor gamma chain (GPVI/FcR gamma chain) signaling pathway.1,2 In this study, we hypothesized that PECAM-1 may be physically and functionally associated with Fc gamma RIIa on the platelet membrane. The functional relationship between PECAM-1 and Fc gamma RIIa was assessed by determining the effect of anti-PECAM-1 monoclonal antibody Fab fragments on Fc gamma RIIa-mediated platelet aggregation and heparin-induced thrombocytopenia (HITS)-mediated platelet aggregation. Preincubation of washed platelets with monoclonal antibody fragments of 2BD4 directed against PECAM-1 and IV.3 directed against Fc gamma RIIa completely blocked Fc gamma RIIa-mediated platelet aggregation and HITS-mediated platelet aggregation, whereas anti-CD151 antibody had no blocking effect. Coengagement of Fc gamma RIIa and PECAM-1 resulted in negative regulation of Fc gamma RIIa-mediated phospholipase C gamma 2 activation, calcium mobilization, and phosphoinositide 3-kinase-dependent signaling pathways. In addition, the physical proximity of Fc gamma RIIa and PECAM-1 was confirmed by using fluorescence resonance energy transfer and coimmunoprecipitation studies. These results indicate that PECAM-1 and Fc gamma RIIa are colocalized on the platelet membrane and PECAM-1 down-regulates Fc gamma RIIa-mediated platelet responses. PMID- 12893768 TI - In human B cells, IL-12 triggers a cascade of molecular events similar to Th1 commitment. AB - Two functionally distinct subsets of B cells that produce Th1- and Th2-like patterns of cytokines have recently been identified. Interleukin-12 (IL-12) is a critical immunoregulatory cytokine that promotes Th1 differentiation through activation of signal transducer and activator of transcription 4 (STAT4). IL-12 has been reported to induce interferon gamma (IFN-gamma) production in B cells, but the relevant signaling pathways are poorly documented. Here, in human primary B cells, we found a functional IL-12 receptor (IL-12R) that internalizes following IL-12 binding. IFN-gamma and, to a lesser extent, IL-12 positively regulated the IL-12Rbeta2 subunit but had no effect on IL-12Rbeta1. On examining the effect of IL-12 on STAT4 and T-bet (2 key factors involved in IFN-gamma promoter activation), we found that IL-12 induced the phosphorylation and nuclear translocation of STAT4. IL-12-dependent constitutive STAT4 activation was also observed in the Epstein-Barr virus (EBV)-transformed B-cell line RPMI 8866 that spontaneously produces IL-12. T-bet expression has been shown to be dependent on STAT1. IL-12 had no direct effect on STAT1 activation or T-bet expression in primary B cells. In contrast, IL-12-induced IFN-gamma led to STAT1 activation, strong expression of T-bet, and IFN-gamma expression. IL-12 therefore initiates a cascade of events in B cells, including STAT4 activation, IL-12Rbeta2 up regulation, IFN-gamma production, and T-bet up-regulation, potentially leading to Th1-like differentiation. PMID- 12893769 TI - Pivotal role of granulocyte colony-stimulating factor in the development of progenitors in the common myeloid pathway. AB - Granulocyte colony-stimulating factor (G-CSF) is the principal cytokine regulating granulopoiesis. G-CSF receptor-deficient mice (G-CSFR-/-) are neutropenic but have only a modest reduction of committed myeloid progenitors. Since it is likely that compensatory mechanisms are induced by the severe neutropenia present in G-CSFR-/- mice, a competitive repopulation assay was performed. These data show that under basal conditions, G-CSF drives nearly all of granulopoiesis through multiple mechanisms. Most importantly, G-CSFR signals regulate the production and/or maintenance of committed-myeloid progenitors. Surprisingly, G-CSFR signals also play a significant role in the regulation of primitive multipotential progenitors in vivo. The contribution of G-CSFR-/- cells to the hematopoietic stem cell compartment is modestly reduced. Moreover, a marked decrease in the contribution of G-CSFR-/- cells to other progenitors in the myeloid pathway, including erythroid and megakaryocytic progenitors, is observed. In contrast, relative to the hematopoietic stem cell compartment, the contribution of G-CSFR-/- cells to the lymphoid lineages is increased. These data suggest that G-CSFR signals may play a role in directing the commitment of primitive hematopoietic progenitors to the common myeloid lineage. Thus, regulation of G-CSF levels may provide a mechanism for directing primitive hematopoietic progenitors into the common myeloid lineage in response to environmental stresses. PMID- 12893770 TI - Interleukin-21 inhibits dendritic cell activation and maturation. AB - Interleukin 21 (IL-21) is a newly described cytokine with homology to IL-4 and IL 15. They belong to a cytokine family that uses the common gamma chain for signaling but also have their private high-affinity receptors. Since it is well known that IL-4 modulates differentiation and activation of dendritic cells (DCs), we analyzed effects of IL-21 compared with IL-15 on DC differentiation, maturation, and function. Here we show that DCs generated with granulocyte macrophage colony-stimulating factor (GMCSF) in the presence of IL-21 (IL-21DCs) differentiated into phenotypically and functionally altered DCs characterized by reduced major histocompatibility complex class II (MHCII) expression, high antigen uptake, and low stimulatory capacity for T-cell activation in vitro. Additionally, IL-21DCs completely failed to induce antigen (Ag)-specific T-cell mediated contact hypersensitivity. Furthermore, IL-21 blocked lipopolysaccharide (LPS)-induced activation and maturation of DCs, which was not mediated by release of the anti-inflammatory cytokine IL-10. In contrast, when supplementing GMCSF with IL-15, DCs differentiated into mature antigen-presenting cells (APCs) with low antigen uptake and highly significant increased capacities to stimulate T cells in vitro and in vivo. Taken together, these results identify a dichotomous action of these structurally related cytokines on DCs, establishing IL-21 as inhibitory cytokine on DC activation and IL-15 as potent stimulator of DC function, making both cytokines interesting targets for therapeutic manipulation of DC-induced immune reactions. PMID- 12893771 TI - Lack of alpha 2-antiplasmin promotes re-endothelialization via over-release of VEGF after vascular injury in mice. AB - We here report that the arterial blood flow after endothelial injury in mice deficient in alpha 2-antiplasmin (alpha 2-AP-/- mice) was well maintained compared with that of wild-type mice. Moreover, the development of neointima 4 weeks after injury in alpha 2-AP-/- mice was significantly decreased. Histologic observations showed a prompt recovery of endothelial cells with a much higher proliferating index in repaired endothelium in alpha 2-AP-/- mice. The amount of secreted vascular endothelial growth factor (VEGF) by explanted vascular smooth muscle cells (SMCs) from alpha 2-AP-/- mice was significantly increased. In separate experiments using a human endothelial cell (EC) line, we could demonstrate that plasminogen binds to ECs and that this binding can be prevented by alpha 2-AP. Finally, an injection of either an anti-VEGF receptor-1 antibody or alpha 2-AP reduced the prompt endothelial healing. alpha 2-AP is the main inactivator of plasmin, which cleaves extracellular matrix-bound VEGF to release a diffusible proteolytic fragment. Lack of alpha 2-AP, therefore, could lead to a local over-release of VEGF by the continuously active plasmin in the injured area, which could result in a prompt re-endothelialization after vascular injury. Our results provide new insight into the role of alpha 2-AP and VEGF in the pathogenesis of re-endothelialization following vascular injury. PMID- 12893772 TI - Primary endothelial cells isolated from the yolk sac and para-aortic splanchnopleura support the expansion of adult marrow stem cells in vitro. AB - The embryonic origin and development of hematopoietic and endothelial cells is highly interdependent. We hypothesized that primary endothelial cells from murine yolk sac and para-aortic splanchnopleura (P-Sp) may possess the capacity to expand hematopoietic stem cells (HSCs) and progenitor cells ex vivo. Using Tie2 GFP transgenic mice in combination with fluorochrome-conjugated monoclonal antibodies to vascular endothelial growth factor receptor-2 (Flk1) and CD41, we have successfully isolated pure populations of primary endothelial cells from 9.5 days after coitus (dpc) yolk sac and P-Sp. Adult murine bone marrow Sca-1+c Kit+lin- cells were cocultured with yolk sac or P-Sp Tie2-GFP+Flk-1+CD41- endothelial cell monolayers for 7 days and the total number of nonadherent cells increased 47- and 295-fold, respectively, and hematopoietic progenitor counts increased 9.4- and 11.4-fold, respectively. Both the yolk sac and P-Sp endothelial cell cocultures facilitated long-term (> 6 months) HSC competitive repopulating ability (2.8- to 9.8-fold increases, respectively). These data suggest that 9.5-dpc yolk sac- and P-Sp-derived primary Tie2-GFP+Flk-1+CD41- endothelial cells possess the capacity to expand adult bone marrow hematopoietic progenitor cell and HSC repopulating ability ex vivo. PMID- 12893773 TI - The proteasome inhibitor bortezomib interacts synergistically with histone deacetylase inhibitors to induce apoptosis in Bcr/Abl+ cells sensitive and resistant to STI571. AB - Interactions between the proteasome inhibitor bortezomib and histone deacetylase inhibitors (HDIs) have been examined in Bcr/Abl+ human leukemia cells (K562 and LAMA 84). Coexposure of cells (24-48 hours) to minimally toxic concentrations of bortezomib + either suberoylanilide hydroxamic acid (SAHA) or sodium butyrate (SB) resulted in a striking increase in mitochondrial injury, caspase activation, and apoptosis, reflected by caspases-3 and -8 cleavage and poly(adenosine diphosphate-ribose) polymerase (PARP) degradation. These events were accompanied by down-regulation of the Raf-1/mitogen-induced extracellular kinase (MEK)/extracellular signal-related kinase (ERK) pathway as well as diminished expression of Bcr/Abl and cyclin D1, cleavage of p21CIP1 and phosphorylation of the retinoblastoma protein (pRb), and induction of the stress-related kinases Jun kinase (JNK) and p38 mitogen-activated protein kinase (MAPK). Transient transfection of cells with a constitutively active MEK construct significantly protected them from bortezomib/SAHA-mediated lethality. Coadministration of bortezomib and SAHA resulted in increased reactive oxygen species (ROS) generation and diminished nuclear factor kappa B (NF-kappa B) activation; moreover, the free radical scavenger L-N-acetylcyteine (LNAC) blocked bortezomib/SAHA-related ROS generation, induction of JNK and p21CIP1, and apoptosis. Lastly, this regimen potently induced apoptosis in STI571 (imatinib mesylate)-resistant K562 cells and CD34+ mononuclear cells obtained from a patient with STI571-resistant disease, as well as in Bcr/Abl- leukemia cells (eg, HL-60, U937, Jurkat). Together, these findings raise the possibility that combined proteasome/histone deacetylase inhibition may represent a novel strategy in leukemia, including apoptosis-resistant Bcr/Abl+ hematologic malignancies. PMID- 12893774 TI - Circadian clock genes oscillate in human peripheral blood mononuclear cells. AB - In mammals, it is well documented that observable circadian rhythms are controlled by a central oscillator that is organized in transcriptional and translational feedback loops involving several clock genes. Although recent studies have demonstrated that clock genes oscillate in many peripheral tissues, their characteristics in the human immune system remain unknown. The present study investigates whether circadian clock genes function in human peripheral blood mononuclear cells. On the basis of studies derived from 3 human subjects under controlled conditions, circadian clock genes hPer1, hPer2, hPer3, and hDec1 are expressed in a circadian manner in human peripheral blood mononuclear cells (PBMCs), with the peak level occurring during the habitual time of activity. The demonstration of functional circadian machinery in human PBMCs suggests that peripheral blood cells may be useful for the investigation of human circadian rhythms and their associated disorders. PMID- 12893775 TI - Transcription from an upstream promoter controls methylation signaling from an inverted repeat of endogenous genes in Arabidopsis. AB - In plants, replication of RNA viruses and RNA from highly transcribed transgenes can trigger DNA methylation. These systems accumulate diced small RNA(smRNA) products of double-stranded RNA(dsRNA) precursors, but it is not known which RNA species directs methylation. The methylated PAI tryptophan biosynthetic genes in Arabidopsis allow the study of methylation signals for endogenous genes with lower expression levels. The PAI genes are arranged as a tandem inverted repeat plus two singlet genes at unlinked loci. Here we show that the predominant PAI transcript initiates at a novel unmethylated promoter that lies upstream of one of the inverted repeat PAI genes. Suppressed transcription from the upstream promoter using transgene-directed silencing reduces methylation on the singlet PAI genes, but not on the inverted repeat, consistent with an RNA methylation signal. RNA gel blots detect normal PAI transcripts and dsRNA read-through species, but not diced smRNAs, suggesting that either precursor dsRNAs or subdetectable levels of smRNAs, below the threshold to effectively degrade PAI transcripts, serve as the PAI methylation signal. Thus, the lower expression endogenous gene system allows dissection of a RNA-directed methylation pathway distinct from RNA degradation pathways. PMID- 12893776 TI - Control of cell number by Drosophila FOXO: downstream and feedback regulation of the insulin receptor pathway. AB - The Drosophila insulin receptor (dInR) regulates cell growth and proliferation through the dPI3K/dAkt pathway, which is conserved in metazoan organisms. Here we report the identification and functional characterization of the Drosophila forkhead-related transcription factor dFOXO, a key component of the insulin signaling cascade. dFOXO is phosphorylated by dAkt upon insulin treatment, leading to cytoplasmic retention and inhibition of its transcriptional activity. Mutant dFOXO lacking dAkt phosphorylation sites no longer responds to insulin inhibition, remains in the nucleus, and is constitutively active. dFOXO activation in S2 cells induces growth arrest and activates two key players of the dInR/dPI3K/dAkt pathway: the translational regulator d4EBP and the dInR itself. Induction of d4EBP likely leads to growth inhibition by dFOXO, whereas activation of dInR provides a novel transcriptionally induced feedback control mechanism. Targeted expression of dFOXO in fly tissues regulates organ size by specifying cell number with no effect on cell size. Our results establish dFOXO as a key transcriptional regulator of the insulin pathway that modulates growth and proliferation. PMID- 12893777 TI - Epithelial cancer in Fanconi anemia complementation group D2 (Fancd2) knockout mice. AB - Fanconi anemia (FA) is a genetic disorder characterized by hypersensitivity to DNA damage, bone marrow failure, congenital defects, and cancer. To further investigate the in vivo function of the FA pathway, mice with a targeted deletion in the distally acting FA gene Fancd2 were created. Similar to human FA patients and other FA mouse models, Fancd2 mutant mice exhibited cellular sensitivity to DNA interstrand cross-links and germ cell loss. In addition, chromosome mispairing was seen in male meiosis. However, Fancd2 mutant mice also displayed phenotypes not observed in other mice with disruptions of proximal FA genes. These include microphthalmia, perinatal lethality, and epithelial cancers, similar to mice with Brca2/Fancd1 hypomorphic mutations. These additional phenotypes were not caused by defects in the ATM-mediated S-phase checkpoint, which was intact in primary Fancd2 mutant fibroblasts. The phenotypic overlap between Fancd2-null and Brca2/Fancd1 hypomorphic mice is consistent with a common function for both proteins in the same pathway, regulating genomic stability. PMID- 12893779 TI - Sizing up the heart: development redux in disease. PMID- 12893778 TI - Mechanism of p38 MAP kinase activation in vivo. AB - The p38 mitogen-activated protein kinase (MAPK) is activated in vitro by three different protein kinases: MKK3, MKK4, and MKK6. To examine the relative roles of these protein kinases in the mechanism of p38 MAP kinase activation in vivo, we examined the effect of disruption of the murine Mkk3, Mkk4, and Mkk6 genes on the p38 MAPK signaling pathway. We show that MKK3 and MKK6are essential for tumor necrosis factor-stimulated p38 MAPK activation. In contrast, ultraviolet radiation-stimulated p38 MAPK activation was mediated by MKK3, MKK4, and MKK6. Loss of p38 MAPK activation in the mutant cells was associated with defects in growth arrest and increased tumorigenesis. These data indicate that p38 MAPK is regulated by the coordinated and selective actions of three different protein kinases in response to cytokines and exposure to environmental stress. PMID- 12893780 TI - The Fanconi road to cancer. PMID- 12893781 TI - The internet and mental health practice. PMID- 12893782 TI - Which treatments are effective for cognitive, behavioural and psychological symptoms in dementia? PMID- 12893783 TI - The number needed to treat: problems describing non-significant results. PMID- 12893785 TI - Review: lifetime prevalence of schizophrenia and related disorders is about 5.5 per 1000, but there is significant variation between regions. PMID- 12893784 TI - About 3 in every 1000 US children in large metropolitan areas may have autism or related developmental disorders. PMID- 12893786 TI - Review: there is marked socioeconomic inequality in persistent depression. PMID- 12893787 TI - Review: black people are more likely than white people to be detained in psychiatric wards in the United Kingdom. PMID- 12893788 TI - Review: antenatal screening tools to predict postnatal depression generally have low specificity and sensitivity. PMID- 12893789 TI - Local adaptation of guidelines for identifying mental disorders in primary care may not improve patient outcomes. PMID- 12893790 TI - Review: atypical antipsychotics and psychosocial interventions, alone or in combination, may reduce youth aggression. PMID- 12893791 TI - Review: it is unclear whether treating internalising disorders in childhood prevents substance abuse in adolescence. PMID- 12893792 TI - Review: parent training and community-based interventions may benefit children with disruptive behaviour disorders. PMID- 12893793 TI - Problem solving programme implemented by teachers may prevent depression in the short term, but longer term benefits are unclear. PMID- 12893794 TI - Review: electroconvulsive therapy may be an effective short term treatment for people with depression. PMID- 12893795 TI - Review: continuing antidepressants for 12 months following acute treatment may reduce the risk of relapse in depression. PMID- 12893796 TI - Adjunctive cognitive therapy is more effective, but more costly, than standard clinical management for relapse prevention in depression. PMID- 12893797 TI - Collaborative care may improve depression management in older adults. PMID- 12893798 TI - Review: counselling in primary care may have short term benefits for people with common mental disorders. PMID- 12893799 TI - Advance directives about treatment preferences had little impact on compulsory readmissions for people with serious mental illness. PMID- 12893800 TI - Review: women with schizophrenia have poorer pregnancy outcomes than other women, but it is unclear whether antipsychotic medications affect their infants. PMID- 12893801 TI - The benefits of exposure therapy alone may last longer than sertraline alone or sertraline plus exposure therapy in social phobia. PMID- 12893802 TI - Hydroxyzine may be safe and effective in generalised anxiety disorder. PMID- 12893804 TI - Clozapine may be more effective than olanzapine for reducing suicidal behaviour in people with schizophrenia at high risk. PMID- 12893803 TI - Buprenorphine may be more effective than clonidine and other symptomatic medications for short term ambulatory heroin withdrawal. PMID- 12893805 TI - Review: cholinesterase inhibitors have a modest effect on neuropsychiatric and functional outcomes in Alzheimer's disease. PMID- 12893806 TI - Ethnicity may be linked to thin body preoccupation and social pressure in the development of eating disorders. PMID- 12893807 TI - People with schizophrenia believe that they are stigmatised at work and in the community. PMID- 12893809 TI - Myosin VI: two distinct roles in endocytosis. AB - Actin is found at the cortex of the cell where endocytosis occurs, but does it play a role in this essential process? Recent studies on the unconventional myosin, myosin VI, an actin-based molecular motor, provide compelling evidence that this myosin and therefore actin is involved in two distinct steps of endocytosis in higher eukaryotes: the formation of clathrin-coated vesicles and the movement of nascent uncoated vesicles from the actin-rich cell periphery to the early endosome. Three distinct adapter proteins--GIPC, Dab2 and SAP97--that associate with the cargo-binding tail domain of myosin VI have been identified. These proteins may recruit myosin VI to its sites of action. PMID- 12893810 TI - Immune semaphorins: a new area of semaphorin research. AB - The semaphorin family comprises soluble and membrane-bound proteins originally identified as axonal guidance cues that function during neuronal development. Emerging evidence suggests that a subset of semaphorins, called 'immune semaphorins', function in the immune system. The class IV semaphorins Sema4D/CD100 and Sema4A use CD72 and Tim-2, respectively, as receptors during immune responses; these receptors comprise a set distinct from those used by semaphorins in the nervous system. Sema4D/CD100, which is expressed constitutively by T cells, is involved in the activation of B cells and dendritic cells, whereas Sema4A is preferentially expressed on B cells and dendritic cells, and is involved in the activation of T cells. Additionally, increasing evidence suggests that some other semaphorins, including viral-encoded semaphorins, might also play important roles in the immune system. PMID- 12893811 TI - Leading the way: Directional sensing through phosphatidylinositol 3-kinase and other signaling pathways. AB - Chemoattractant-responsive cells are able to translate a shallow extracellular chemical gradient into a steep intracellular gradient resulting in the localization of F-actin assembly at the front and an actomyosin network at the rear that moves the cell forward. Recent evidence suggests that one of the first asymmetric cellular responses is the localized accumulation of PtdIns(3,4,5)P3, the product of class I phosphoinositide 3-kinase (PI3K) at the site of the new leading edge. The strong accumulation of PtdIns(3,4,5)P3 results from the localized activation of PI3K and also from feedback loops that amplify PtdIns(3,4,5)P3 synthesis at the front and control its degradation at the side and back of cells. These different pathways are temporally and spatially regulated and integrate with other signaling pathways during directional sensing and chemotaxis. PMID- 12893812 TI - ECM-induced gap junctional communication enhances mammary epithelial cell differentiation. AB - The relationship between gap junctional intercellular communication (GJIC) and mammary cell (CID-9) differentiation in vitro was explored. CID-9 cells differentiate and express beta-casein in an extracellular matrix (ECM)- and hormone-dependent manner. In response to interaction with the ECM, cells in culture modulated the expression of their gap junction proteins at the transcriptional and post-translational levels. In the presence of EHS-matrix, connexins (Cx)26, 32 and 43 localized predominantly to the plasma membrane, and enhanced GJIC [as measured by Lucifer Yellow (LY) dye transfer assays] was noted. Inhibition of GJIC of cells on EHS-matrix with 18 alpha glycyrrhetinic acid (GA) resulted in reversible downregulation of beta-casein expression. In the presence of cAMP, cells cultured on plastic expressed beta-casein, upregulated Cx43 and Cx26 protein levels and enhanced GJIC. This was reversed in the presence of 18 alpha GA. cAMP-treated cells plated either on a non-adhesive PolyHEMA substratum or on plastic supplemented with function-blocking anti-beta 1 integrin antibodies, maintained beta-casein expression. These studies suggest that cell ECM interaction alone may induce differentiation through changes in cAMP levels and formation of functional gap junctions. That these events are downstream of ECM signalling was underscored by the fact that enhanced GJIC induced partial differentiation in mammary epithelial cells in the absence of an exogenously provided basement membrane and in a beta 1-integrin- and adhesion-independent manner. PMID- 12893813 TI - Drosophila Rheb GTPase is required for cell cycle progression and cell growth. AB - Precise body and organ sizes in the adult animal are ensured by a range of signaling pathways. In a screen to identify genes affecting hindgut morphogenesis in Drosophila, we identified a P-element insertion in dRheb, a novel, highly conserved member of the Ras superfamily of G-proteins. Overexpression of dRheb in the developing fly (using the GAL4:UAS system) causes dramatic overgrowth of multiple tissues: in the wing, this is due to an increase in cell size; in cultured cells, dRheb overexpression results in accumulation of cells in S phase and an increase in cell size. Using a loss-of-function mutation we show that dRheb is required in the whole organism for viability (growth) and for the growth of individual cells. Inhibition of dRheb activity in cultured cells results in their arrest in G1 and a reduction in size. These data demonstrate that dRheb is required for both cell growth (increase in mass) and cell cycle progression; one explanation for this dual role would be that dRheb promotes cell cycle progression by affecting cell growth. Consistent with this interpretation, we find that flies with reduced dRheb activity are hypersensitive to rapamycin, an inhibitor of the growth regulator TOR. In cultured cells, the effect of overexpressing dRheb was blocked by the addition of rapamycin. These results imply that dRheb is involved in TOR signaling. PMID- 12893814 TI - Moments of discovery: my favorite experiments. PMID- 12893815 TI - Toll-like receptor signaling. PMID- 12893816 TI - Unfolding the action of progesterone receptors. PMID- 12893817 TI - beta-Amyloid induces paired helical filament-like tau filaments in tissue culture. AB - Paired helical filaments (PHF) are the principal pathologic components of neurofibrillary tangles in Alzheimer's disease (AD). To reproduce the formation of PHF in tissue culture, we stably expressed human tau with and without pathogenic mutations in human SH-SY5Y cells and exposed them for 5 days to aggregated synthetic beta-amyloid peptide (A beta 42). This caused a decreased solubility of tau along with the generation of PHF-like tau-containing filaments. These were 20 nm wide and had periodicities of 130-140 nm in the presence of P301L mutant tau or 150-160 nm in the presence of wild-type tau. Mutagenesis of the phosphoepitope serine 422 of tau prevented both the A beta 42-mediated decrease in solubility and the generation of PHF-like filaments, suggesting a role of serine 422 or its phosphorylation in tau filament formation. Together, our data underscore a role of A beta 42 in the formation of PHF-like filaments. Our culture system will be useful to map phosphoepitopes of tau involved in PHF formation and to identify and characterize modifiers of the tau pathology. Further adaptation of the system may allow the screening and validation of compounds designed to prevent PHF formation. PMID- 12893818 TI - Identification and characterization of E2F7, a novel mammalian E2F family member capable of blocking cellular proliferation. AB - The mammalian E2F family of transcription factors plays a crucial role in the regulation of cellular proliferation, apoptosis, and differentiation. Consistent with its biological role in a number of important cellular processes, E2F regulates the expression of genes involved in cell cycle, DNA replication, DNA repair, and mitosis. It has proven difficult, however, to determine the specific roles played by the various known family members in these cellular processes. The work presented here now extends the complexity of this family even further by the identification of a novel E2F family member, which we now term E2F7. Like the expression of the known E2F activators, E2F1, E2F2, and E2F3, the expression of E2F7 is growth-regulated, at least in part, through E2F binding elements on its promoter, and its protein product is localized to the nucleus and associates with DNA E2F recognition sites with high affinity. A number of salient features, however, make this member unique among the E2F family. First, the E2F7 gene encodes a protein that possesses two distinct DNA-binding domains and that lacks a dimerization domain as well as a transcriptional activation and a retinoblastoma-binding domain. In contrast to the E2F activators, E2F7 can block the E2F-dependent activation of a subset of E2F target genes as well as mitigate cellular proliferation of mouse embryo fibroblasts. These findings identify E2F7 as a novel member of the mammalian E2F transcription factor family that has properties of a transcriptional repressor capable of negatively influencing cellular proliferation. PMID- 12893819 TI - Glucose metabolism in cancer: importance of transcription factor-DNA interactions within a short segment of the proximal region og the type II hexokinase promoter. AB - A common signature of many cancers is a high glucose catabolic rate frequently dependent on the overexpression of Type II hexokinase (HKII), a mitochondrial bound enzyme that also suppresses cell death. As the tumor HKII promoter plays a significant role in HKII overexpression, studies reported here were undertaken to identify both the major regions and transcription factors involved under tumor like conditions. Reporter gene assays following transfection of hepatoma cells with decreasing segments of the HKII promoter traced its known strength to the proximal region (-281 to -35). Mutational analyses showed that in this short region GC boxes 1, 2, 5, and 6, a CCAAT box, an inverted CCAAT box, and CRE are involved in promoter activation. Other studies demonstrated binding of transcription factors Sp1, Sp2, and Sp3 to GC boxes 1 and 6, Sp1 and Sp2 to GC boxes 2 and 5, NF-Y to CCAAT boxes, and CREB, ATF1, and CREM to CRE. In addition, transfection studies involving Sp1, Sp2, Sp3, CREB, and NFY (dominant negative form) provided evidence that these proteins are promoter activators. Finally, alignment of available HK proximal promoters showed strong conservation only among HKII sequences. These findings implicate signaling pathways directed to a short segment of the proximal region of the HKII promoter as major contributors to HKII overexpression in many cancers. PMID- 12893820 TI - Mutations in alpha-chain of C4BP that selectively affect its factor I cofactor function. AB - C4b-binding protein (C4BP) inhibits all pathways of complement activation, acting as a cofactor to the serine protease factor I (FI) in the degradation of activated complement factors C4b and C3b. C4BP is a disulfide-linked polymer of seven alpha-chains and a unique beta-chain, the alpha- and beta-chains being composed of eight and three complement control protein (CCP) domains, respectively. In previous studies we have localized cofactor activity and binding of C4b to alpha-chain CCP1-3 of C4BP, whereas the binding of C3b required additionally CCP4. Likewise, introduced point mutations that decreased binding of C4b/C3b caused a decrease in cofactor activity. In the present study, we describe two mutants of C4BP, K126Q/K128Q and F144S/F149S, clustered on alpha-chain CCP3, which selectively lost their ability to act as cofactors in the cleavage of both C4b and C3b. Both mutants show the same binding affinity for C4b/C3b as measured by surface plasmon resonance and have the same inhibitory effect on formation and decay of the classical pathway C3-convertase as the wild type C4BP. It appears that C4b and C3b do not undergo the same conformational changes upon binding to the C4BP mutants as during the interaction with the wild type C4BP, which then results in the observed loss of the cofactor activity. PMID- 12893821 TI - Differential regulation of the muscle-specific GLUT4 enhancer in regenerating and adult skeletal muscle. AB - We have reported a novel functional co-operation among MyoD, myocyte enhancer factor-2 (MEF2), and the thyroid hormone receptor in a muscle-specific enhancer of the rat GLUT4 gene in muscle cells. Here, we demonstrate that the muscle specific enhancer of the GLUT4 gene operates in skeletal muscle and is muscle fiber-dependent and innervation-independent. Under normal conditions, both in soleus and in extensor digitorum longus muscles, the activity of the enhancer required the integrity of the MEF2-binding site. Cancellation of the binding site of thyroid hormone receptor enhanced its activity, suggesting an inhibitory role. Muscle regeneration of the soleus and extensor digitorum longus muscles caused a marked induction of GLUT4 and stimulation of the enhancer activity, which was independent of innervation. During muscle regeneration, the enhancer activity was markedly inhibited by cancellation of the binding sites of MEF2, MyoD, or thyroid hormone receptors. Different MEF2 isoforms expressed in skeletal muscle (MEF2A, MEF2C, and MEF2D) and all members of the MyoD family had the capacity to participate in the activity of the GLUT4 enhancer as assessed by transient transfection in cultured cells. Our data indicate that the GLUT4 enhancer operates in muscle fibers and its activity contributes to the differences in GLUT4 gene expression between oxidative and glycolytic muscle fibers and to the GLUT4 up-regulation that occurs during muscle regeneration. The activity of the enhancer is maintained in adult muscle by MEF2, whereas during regeneration the operation of the enhancer depends on MEF2, myogenic transcription factors of the MyoD family, and thyroid hormone receptors. PMID- 12893822 TI - v-SRC specifically regulates the nucleo-cytoplasmic delocalization of the major isoform of TEL (ETV6). AB - TEL is a frequent target of chromosomal translocations in human cancer and an alleged tumor suppressor gene. TEL encodes two isoforms: a major TEL-M1 isoform as well as TEL-M43, which lacks the first 42 amino acid residues of TEL-M1. Both isoforms are potent transcriptional repressors that can inhibit RAS-induced transformation. Here we show that the v-SRC protein-tyrosine kinase relieves the repressive activity of TEL-M1, an activity that is associated with the v-SRC induced delocalization of TEL-M1 from the nucleus to the cytoplasm. TEL-M1 delocalization requires the kinase activity of v-SRC and is not induced by oncogenic RAS or AKT. Cytoplasmic delocalization of TEL-M1 in response to v-SRC critically depends upon its unique amino-terminal domain (SRCD domain) because (i). v-SRC did not inhibit the repressive properties of TEL-M43, nor affected TEL M43 nuclear localization; (ii). fusion of the first 52 amino acid residues of TEL M1 to FLI-1, an ETS protein insensitive to v-SRC-induced delocalization, is sufficient to confer v-SRC-induced delocalization to this TEL/FLI-1 chimeric protein. The v-SRC-induced nucleo-cytoplasmic delocalization of TEL-M1 does not involve phosphorylation of the SRCD and does not require TEL self-association and repressive domains. Finally, enforced expression of the v-SRC-insensitive TEL M43, but not of TEL-M1, inhibits v-SRC-induced transformation of NIH3T3 fibroblasts. These results identify a regulatory domain in TEL that specifically impinges on the subcellular localization of its major TEL-M1 isoform. They, furthermore, indicate that inhibition of TEL-M1 nuclear function is required for v-SRC to induce cellular transformation. PMID- 12893823 TI - Bacteriophage T4Dam (DNA-(adenine-N6)-methyltransferase): evidence for two distinct stages of methylation under single turnover conditions. AB - We compared the (pre)steady-state and single turnover methylation kinetics of bacteriophage T4Dam (DNA-(adenine-N6)-methyltransferase)-mediated methyl group transfer from S-adenosyl-l-methionine (AdoMet) to oligodeoxynucleotide duplexes containing a single recognition site (palindrome 5'-GATC/5'-GATC) or some modified variant. T4Dam-AdoMet functions as a monomer under steady-state conditions (enzyme/DNA << 1), whereas under single turnover conditions (enzyme/DNA > 1), a catalytically active complex containing two Dam-AdoMet molecules is formed initially, and two methyl groups are transferred per duplex (to produce a methylated duplex and S-adenosyl-l-homocysteine (AdoHcy)). We propose that the single turnover reaction proceeds in two stages. First, two preformed T4Dam-AdoMet complexes bind opposite strands of the unmodified target site, and one enzyme molecule catalyzes the rapid transfer of the AdoMet-methyl group (kmeth1 = 0.21 s-1); this is 2.5-fold slower than the rate observed with monomeric T4Dam-AdoMet bound under pre-steady-state conditions for burst determination. In the second stage, methyl transfer to adenine in GATC on the complementary strand occurs at a rate that is 1 order of magnitude slower (kmeth2 = 0.023 s-1). We suggest that under single turnover conditions, methylation of the second strand is rate-limited by Dam-AdoHcy dissociation or its clearance from the methylated complementary strand. The hemimethylated duplex 5'-GATC/5' GMTC also interacts with T4Dam-AdoMet complexes in two stages under single turnover reaction conditions. The first stage (kmeth1) reflects methylation by dimeric T4Dam-AdoMet productively oriented to the strand with the adenine residue capable of methylation. The slower second stage (kmeth2) reflects methylation by enzyme molecules non-productively oriented to the GMTC chain, which then have to re-orient to the opposite productive chain. Substitutions of bases and deletions in the recognition site affect the kinetic parameters in different fashions. When the GAT portion of GATC was disrupted, the proportion of the initial productive enzyme-substrate complexes was sharply reduced. PMID- 12893824 TI - Glutathione peroxidase 1 is regulated by the c-Abl and Arg tyrosine kinases. AB - The c-Abl and Arg tyrosine kinases are activated in the cellular response to oxidative stress. The present studies demonstrate that c-Abl and Arg associate with glutathione peroxidase 1 (GPx1) and that this interaction is regulated by intracellular oxidant levels. The c-Abl and Arg SH3 domains bind directly to a proline-rich site in GPx1 at amino acids 132-145. GPx1 also functions as a substrate for c-Abl- and Arg-mediated phosphorylation on Tyr-96. The results further show that c-Abl and Arg stimulate GPx activity and that these kinases contribute to GPx-mediated protection of cells against oxidative stress. Our findings provide the first evidence that GPx1 is regulated by a signaling pathway that is activated in the oxidative stress response. PMID- 12893825 TI - Transforming growth factor-beta-mediated chondrogenesis of human mesenchymal progenitor cells involves N-cadherin and mitogen-activated protein kinase and Wnt signaling cross-talk. AB - The multilineage differentiation potential of adult tissue-derived mesenchymal progenitor cells (MPCs), such as those from bone marrow and trabecular bone, makes them a useful model to investigate mechanisms regulating tissue development and regeneration, such as cartilage. Treatment with transforming growth factor beta (TGF-beta) superfamily members is a key requirement for the in vitro chondrogenic differentiation of MPCs. Intracellular signaling cascades, particularly those involving the mitogen-activated protein (MAP) kinases, p38, ERK-1, and JNK, have been shown to be activated by TGF-betas in promoting cartilage-specific gene expression. MPC chondrogenesis in vitro also requires high cell seeding density, reminiscent of the cellular condensation requirements for embryonic mesenchymal chondrogenesis, suggesting common chondro-regulatory mechanisms. Prompted by recent findings of the crucial role of the cell adhesion protein, N-cadherin, and Wnt signaling in condensation and chondrogenesis, we have examined here their involvement, as well as MAP kinase signaling, in TGF beta1-induced chondrogenesis of trabecular bone-derived MPCs. Our results showed that TGF-beta1 treatment initiates and maintains chondrogenesis of MPCs through the differential chondro-stimulatory activities of p38, ERK-1, and to a lesser extent, JNK. This regulation of MPC chondrogenic differentiation by the MAP kinases involves the modulation of N-cadherin expression levels, thereby likely controlling condensation-like cell-cell interaction and progression to chondrogenic differentiation, by the sequential up-regulation and progressive down-regulation of N-cadherin. TGF-beta1-mediated MAP kinase activation also controls WNT-7A gene expression and Wnt-mediated signaling through the intracellular beta-catenin-TCF pathway, which likely regulates N-cadherin expression and subsequent N-cadherin-mediated cell-adhesion complexes during the early steps of MPC chondrogenesis. PMID- 12893826 TI - Sensitivity of Caenorhabditis elegans clk-1 mutants to ubiquinone side-chain length reveals multiple ubiquinone-dependent processes. AB - Ubiquinone (coenzyme Q, or Q) is a membrane constituent, whose head group is capable of accepting and donating electrons and whose lipidic side chain is composed of a variable number of isoprene subunits. A possible role for Q as a dietary antioxidant for treating conditions that involve altered cellular redox states is being intensely studied. Mutations in the clk-1 gene of the nematode Caenorhabditis elegans affect numerous physiological rates including behavioral rates, developmental rates, reproduction, and life span. clk-1 encodes a protein associated with the inner mitochondrial membrane that is necessary for Q biosynthesis in C. elegans. clk-1 mutants do not synthesize Q but accumulate demethoxyubiquinone, a Q synthesis intermediate that is able to partially sustain mitochondrial respiration in worms as well as in mammals. Recently, we and others have found that exogenous Q is necessary for the fertility and development of clk 1 mutants. Here, we take advantage of the clk-1 genetic model to identify structural features of Q that are functionally important in vivo. We show that clk-1 mutants are exquisitely sensitive to the length of the side chain of the Q they consume. We also identified differential sensitivity to Q side-chain length between null alleles of clk-1 (qm30 and qm51) and the weaker allele e2519. This allows us to propose a model where we distinguish several types of Q-dependent processes in vivo: processes that are very sensitive to Q side-chain length and processes that are permissive to Q with shorter chains. PMID- 12893827 TI - Amyloid beta protein precursor (AbetaPP), but not AbetaPP-like protein 2, is bridged to the kinesin light chain by the scaffold protein JNK-interacting protein 1. AB - Proteolytic processing of amyloid beta protein precursor (AbetaPP) generates peptides that regulate normal cell signaling and are implicated in Alzheimer's disease pathogenesis. AbetaPP processing also occurs in nerve processes where AbetaPP is transported from the cell body by kinesin-I, a microtubule motor composed of two kinesin heavy chain and two kinesin light chain (Klc) subunits. AbetaPP transport is supposedly mediated by the direct AbetaPP-Klc1 interaction. Here we demonstrate that the AbetaPP-Klc1 interaction is not direct but is mediated by JNK-interacting protein 1 (JIP1). The phosphotyrosine binding domain of JIP1 binds the cytoplasmic tail of AbetaPP, whereas the JIP1 C-terminal region interacts with the tetratrico-peptide repeats of Klc1. We also show that JIP1 does not bridge the AbetaPP gene family member AbetaPP-like protein 2, APLP2, to Klc1. These results support a model where JIP1 mediates the interaction of AbetaPP to the motor protein kinesin-I and that this JIP1 function is unique for AbetaPP relative to its family member APLP2. Our data suggest that kinesin-I dependent neuronal AbetaPP transport, which controls AbetaPP processing, may be regulated by JIP1. PMID- 12893828 TI - Human erythrocyte membrane band 3 protein influences hemoglobin cooperativity. Possible effect on oxygen transport. AB - Hemoglobin function can be modulated by the red cell membrane but some mechanistic details are incomplete. For example, the 43-kDa chymotryptic fragment of the cytoplasmic portion of red cell membrane Band 3 protein and its corresponding N-terminal 11-residue synthetic peptide lower the oxygen affinity of hemoglobin but effects on cooperativity are unclear. Using highly purified preparations, we also find a lowered Hill coefficient (n values <2) at subequivalent ratios of Band 3 fragment or of synthetic peptide to Hb, resulting in an oxygen affinity that is moderately decreased and a partially hyperbolic shape for the O2 binding curve. Both normal HbA and sickle HbS display this property. Thus, the determinant responsible for the Hb cooperativity decreases by the 43-kDa fragment resides within its first 11 N-terminal residues. This effect is observed in the absence of chloride and is reversed by its addition. As effector to Hb ratios approach equivalence or with saturating chloride normal cooperativity is restored, and oxygen affinity is further lowered because the shape of the oxygen binding curve becomes completely sigmoidal. The relative efficiencies of 2,3-diphosphoglycerate (DPG), the 43-kDa Band 3 fragment, and the 11-residue synthetic peptide in lowering cooperativity are very similar. The findings are explained based on the stereochemical mechanism of cooperativity because of two populations of T-state hemoglobin tetramers, one with bound effector and the other with free (Perutz, M. F. (1989) Q. Rev. Biophys. 22, 139 237). As a result of this property, hemoglobin at the membrane inner surface in contact with the N-terminal region of Band 3 could preferentially bind O2 at low oxygen tension and then release it upon saturation with 2,3-diphosphoglycerate in the interior of the red cell. Membrane modulation of hemoglobin oxygen affinity has particularly interesting implications for the polymerization of hemoglobin S in the sickle red cell. PMID- 12893829 TI - A cytosolic domain of the yeast Zrt1 zinc transporter is required for its post translational inactivation in response to zinc and cadmium. AB - Nutrient metals such as zinc are both essential to life and potentially toxic if overaccumulated by cells. Non-essential toxic metals like cadmium can enter cells through the uptake transporters responsible for nutrient metal acquisition. Therefore, in the face of ever changing extracellular metal levels, organisms tightly control their intracellular levels of nutrient metals and prevent accumulation of toxic metals. We show here that post-translational inactivation of the yeast Zrt1 zinc uptake transporter is important for zinc homeostasis. During the transition from zinc-limiting to zinc-replete growth conditions (i.e. zinc shock), the Zrt1 transporter is ubiquitinated, endocytosed, and subsequently degraded in the vacuole. To further understand this process at a molecular level, we mapped a region of Zrt1 required for ubiquitination and endocytosis in response to zinc to a domain located on the intracellular surface of the plasma membrane. This domain is a critical cis-acting component of the metal signaling pathway that controls Zrt1 protein trafficking. Using mutant alleles defective for metal-responsive inactivation, we also show that Zrt1 inactivation may be an important mechanism for preventing cadmium uptake and toxicity in zinc-limited cells. PMID- 12893830 TI - The C2-like beta-barrel domain mediates the Ca2+-dependent resistance of 5 lipoxygenase activity against inhibition by glutathione peroxidase-1. AB - Recently, we reported that in crude enzyme preparations, a monocyte-derived soluble protein (M-DSP) renders 5-lipoxygenase (5-LO) activity Ca2+-dependent. Here we provide evidence that this M-DSP is glutathione peroxidase (GPx)-1. Thus, the inhibitory effect of the M-DSP on 5-LO could be overcome by the GPx-1 inhibitor mercaptosuccinate and by the broad spectrum GPx inhibitor iodoacetate, as well as by addition of 13(S)-hydroperoxy-9Z,11E-octadecadienoic acid (13(S) HPODE). Also, the chromatographic characteristics and the estimated molecular mass (80-100 kDa) of the M-DSP fit to GPx-1 (87 kDa), and GPx-1, isolated from bovine erythrocytes, mimicked the effects of the M-DSP. Intriguingly, only a trace amount of thiol (10 micro M GSH) was required for reduction of 5-LO activity by GPx-1 or the M-DSP. Moreover, the requirement of Ca2+ allowing 5-LO product synthesis in various leukocytes correlated with the respective GPx-1 activities. Mutation of the Ca2+ binding sites within the C2-like domain of 5-LO resulted in strong reduction of 5-LO activity by M-DSP and GPx-1, also in the presence of Ca2+. In summary, our data suggest that interaction of Ca2+ at the C2 like domain of 5-LO protects the enzyme against the effect of GPx-1. Apparently, in the presence of Ca2+, a low lipid hydroperoxide level is sufficient for 5-LO activation. PMID- 12893831 TI - Integrin-mediated invasion of Staphylococcus aureus into human cells requires Src family protein-tyrosine kinases. AB - Staphylococcus aureus, a common cause of nosocomial infections, is able to invade eukaryotic cells by indirectly engaging beta1 integrin-containing host receptors, whereas non-pathogenic Staphylococcus carnosus is not invasive. Here, we identify intracellular signals involved in integrin-initiated internalization of S. aureus. In particular, the host cell actin cytoskeleton and Src family protein tyrosine kinases (PTKs) are essential to mediate S. aureus invasion. Src PTKs are activated in response to pathogenic S. aureus, but not S. carnosus. In addition, pharmacological and genetic interference with Src PTK function reduces bacterial internalization. Importantly, Src PTK-deficient cells are resistant to S. aureus invasion, demonstrating the essentiality of host Src PTKs in integrin-mediated uptake of this pathogen. PMID- 12893832 TI - Vascular endothelial growth factor up-regulates expression of receptor activator of NF-kappa B (RANK) in endothelial cells. Concomitant increase of angiogenic responses to RANK ligand. AB - Vascular endothelial growth factor (VEGF) is known as a key regulator of angiogenesis during endochondral bone formation. Recently, we demonstrated that TNF-related activation-induced cytokine (TRANCE or RANKL), which is essential for bone remodeling, also had an angiogenic activity. Here we report that VEGF up regulates expression of receptor activator of NF-kappa B (RANK) and increases angiogenic responses of endothelial cells to TRANCE. Treatment of human umbilical vein endothelial cells (HUVECs) with VEGF increased both RANK mRNA and surface protein expression. Although placenta growth factor specific to VEGF receptor-1 had no significant effect on RANK expression, inhibition of downstream signaling molecules of the VEGF receptor-2 (Flk-1/KDR) such as Src, phospholipase C, protein kinase C, and phosphatidylinositol 3'-kinase suppressed VEGF-stimulated RANK expression in HUVECs. Moreover, the MEK inhibitor PD98059 or expression of dominant negative MEK1 inhibited induction of RANK by VEGF but not the Ca2+ chelator 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid-acetoxymethyl ester (BAPTA-AM). VEGF potentiated TRANCE-induced ERK activation and tube formation via RANK up-regulation in HUVECs. Together, these results show that VEGF enhances RANK expression in endothelial cells through Flk-1/KDR-protein kinase C-ERK signaling pathway, suggesting that VEGF plays an important role in modulating the angiogenic action of TRANCE under physiological or pathological conditions. PMID- 12893833 TI - Identification and characterization of a novel Pyk2/related adhesion focal tyrosine kinase-associated protein that inhibits alpha-synuclein phosphorylation. AB - alpha-Synuclein is a presynaptic protein involved in the pathogenesis of several neurodegenerative diseases, such as Parkinson's disease. Pyk2/related adhesion focal tyrosine kinase (RAFTK) tyrosine kinase is an upstream regulator of Src family kinases in the central nervous system that is involved in alpha-synuclein phosphorylation. The present study reports the cloning and characterization of a novel adaptor protein, Pyk2/RAFTK-associated protein (PRAP), that specifically binds to Pyk2/RAFTK and inhibits alpha-synuclein tyrosine phosphorylation. PRAP contains a coiled-coil domain, a pleckstrin homology domain, and a SH3 domain; the SH3 domain binds to the proline-rich domain of Pyk2/RAFTK. PRAP was observed to be present throughout the brain, including substantia nigra dopaminergic neurons, in which it localized to the cytoplasm. PRAP was found to function as a substrate for Src family kinases, such as c-Src or Fyn, but not for Pyk2/RAFTK. Hyperosmotic stress induced phosphorylation of tyrosine 125 of alpha-synuclein via Pyk2/RAFTK, which acted through Src family kinases. Such phosphorylation was inhibited by PRAP expression, suggesting that PRAP negatively regulates alpha synuclein phosphorylation following cell stress. In conclusion, PRAP functions as a downstream target for Pyk2/RAFTK and plays a role in alpha-synuclein phosphorylation. PMID- 12893834 TI - Projection structure of the atractyloside-inhibited mitochondrial ADP/ATP carrier of Saccharomyces cerevisiae. AB - ADP/ATP carriers in the inner mitochondrial membrane catalyze the exchange of cytosolic ADP for ATP synthesized in the mitochondrial matrix by ATP synthase and thereby replenish the eukaryotic cell with metabolic energy. The yeast ADP/ATP carrier (AAC3) was overexpressed, inhibited by atractyloside, purified, and reconstituted into two-dimensional crystals. Images of frozen hydrated crystals were recorded by electron microscopy, and a projection structure was calculated to 8-A resolution. The AAC3 molecule has pseudo 3-fold symmetry in agreement with the 3-fold sequence repeats that are typical of members of the mitochondrial carrier family. The density distribution is consistent with a bundle of six transmembrane alpha-helices with two or three short alpha-helical extensions closing the central pore on the matrix side. The AAC3 molecules in the crystal are arranged in symmetrical homo-dimers, but the translocation pore for adenine nucleotides lies in the center of the molecule and not along the dyad axis of the dimer. PMID- 12893835 TI - Biliary secretion of glutathione in estradiol 17beta-D-glucuronide-induced cholestasis. AB - Estradiol-17beta-D-glucuronide (E2-17G) induces an acute but reversible inhibition of bile flow after its intravenous administration to rats, due in part to the endocytic retrieval of the canalicular multidrug resistance-associated transporter protein 2 and the bile salt export pump, transporters that contribute to bile flow. Decreased bile salt-independent bile flow (BSIF) is also involved and persists during the phase of recovery from cholestasis. Because glutathione and HCO3- are major contributors to BSIF, we evaluated changes in their biliary excretion and the hepatic content of total glutathione during E2-17G-induced cholestasis. E2-17G acutely decreased bile flow and biliary excretion of total glutathione by about 80%; glutathione excretion was still inhibited at 80 min and 120 min, even though bile flow was partially and totally restored, respectively. Neither liver glutathione content nor the proportions of oxidized glutathione in bile and liver were affected by E2-17G at any time. HCO3- concentrations in bile were unchanged, so that secretion paralleled variations in bile flow. In the isolated perfused liver, addition of E2-17G decreased both bile flow and the biliary concentration of glutathione, whereas addition of its noncholestatic isomer estradiol-3-D-glucuronide (E2-3G) did not inhibit bile flow, but significantly reduced the concentration of glutathione in bile. The bile:liver concentration ratios of glutathione were significantly decreased in vivo by E2 17G and in the perfused liver by E2-17G and E2-3G. These data indicate that E2 17G cis-inhibits the canalicular transport of glutathione and thus contributes to the cholestatic effect by inhibiting BSIF. PMID- 12893836 TI - Functional expression of the multidrug resistance protein 1 in microglia. AB - Brain expression of the multidrug resistance proteins (MRPs), a collection of membrane-associated ATP-dependent efflux transporters, is poorly understood. Although several studies have examined the expression of these proteins within the brain barriers (i.e., the blood-brain barrier and choroid plexus), little information is available with respect to brain parenchyma cells such as microglia and astrocytes. Because microglia are the primary brain cells infected by the human immunodeficiency virus type 1 (HIV-1), MRP1 expression within microglia may contribute to lower brain accumulation of anti-HIV drugs. To examine the expression pattern of MRP1 within microglia, we performed reverse transcriptase polymerase chain reaction analysis and Western blotting on a rat brain microglia cell line MLS-9, and in primary cultures of rat microglia. Both rat MRP1 (rMPR1) mRNA and protein were expressed in the cell line, as well as the primary cultures. We then characterized rMRP1-mediated transport properties in MLS-9 cells using [3H]vincristine, a known MRP1 substrate. Vincristine accumulation by monolayers of MLS-9 cells increased significantly in the presence of several well established MRP1 inhibitors (MK571, genistein, sulfinpyrazone, probenecid, and indomethacin), protease inhibitors, or the ATPase inhibitor sodium azide. In addition, vincristine accumulation was significantly modulated by altering the intracellular concentration of the reduced form of glutathione, further suggesting the involvement of rMRP1-mediated transport. These results provide strong evidence that the MRP1 protein is both expressed and functional in microglia cells. They also suggest that brain parenchyma can act as a "second" barrier to drug permeability and regulate brain distribution/accumulation of various xenobiotics, including protease inhibitors. PMID- 12893837 TI - Ranitidine treatment during a modest inflammatory response precipitates idiosyncrasy-like liver injury in rats. AB - Drug idiosyncrasy is an adverse event of unknown etiology that occurs in a small fraction of people taking a drug. Some idiosyncratic drug reactions may occur from episodic decreases in the threshold for drug hepatotoxicity. Previous studies in rats have shown that modest underlying inflammation triggered by bacterial lipopolysaccharide (LPS) can decrease the threshold for xenobiotic hepatotoxicity. The histamine-2 (H2)-receptor antagonist ranitidine (RAN) causes idiosyncratic reactions in people, with liver as a usual target. We tested the hypothesis that RAN could be rendered hepatotoxic in animals undergoing a modest inflammatory response. Male rats were treated with a nonhepatotoxic dose of LPS (44 x 10(6) endotoxin units/kg i.v.) or its vehicle and then 2 h later with a nonhepatotoxic dose of RAN (30 mg/kg i.v.) or its vehicle. Liver injury was evident only in animals treated with both RAN and LPS as estimated by increases in serum alanine aminotransferase, aspartate aminotransferase, and gamma-glutamyl transferase activities within 6 h after RAN administration. LPS/RAN cotreatment resulted in midzonal liver lesions characterized by acute necrosuppurative hepatitis. Famotidine (FAM) is an H2-antagonist for which the propensity for idiosyncratic reactions is far less than RAN. Rats given LPS and FAM at a dose pharmacologically equipotent to that of RAN did not develop liver injury. In vitro, RAN sensitized hepatocytes to killing by cytotoxic products from activated neutrophils, whereas FAM lacked this ability. The results indicate that a response resembling human RAN idiosyncrasy can be reproduced in animals by RAN exposure during modest inflammation. PMID- 12893838 TI - 17Beta-estradiol as a receptor-mediated cardioprotective agent. AB - Cardiac tissue that undergoes an ischemic episode exhibits irreversible alterations that become more extensive upon reperfusion. Estrogen treatment has been reported to protect against reperfusion injury, but the mechanism remains unknown. The cardioprotective effects of 17beta-estradiol, a biologically active form of the hormone, and 17alpha-estradiol were assessed in an in vivo occlusion reperfusion model. Anesthetized, ovariectomized rabbits were administered 17beta estradiol (20 microg), 17alpha-estradiol (1 mg), or vehicle intravenously 30 min before a 30-min occlusion of the left anterior descending (LAD) coronary artery followed by 4 h of reperfusion. Infarct size as a percentage of area at risk decreased in the 17beta-estradiol-treated group (18.8 +/- 1.7) compared with 17alpha-estradiol (41.9 +/- 4.8; P < 0.01) or vehicle groups (48 +/- 5.5; P < 0.001). Similar results were obtained when infarct size was expressed as a percentage of total left ventricle. The second objective of the study was to assess fulvestrant (Faslodex, ICI 182,780), an estrogen receptor antagonist, for its effects on infarct size in ovariectomized female rabbits treated with 17beta estradiol. ICI 182,780 was administered intravenously 1 h before the administration of 17beta-estradiol (20 microg) or vehicle. The hearts were subjected to 30-min LAD coronary artery occlusion and 4 h of reperfusion. Pretreatment with ICI 182,780 significantly limited the infarct size sparing effect of 17beta-estradiol when expressed as a percentage of the risk region (53.0 +/- 5.0). The results indicate that 17beta-estradiol protects the heart against ischemia-reperfusion injury and that the observed cardioprotection is mediated by the estrogen receptor. PMID- 12893839 TI - Zeta isoform of protein kinase C prevents oxidant-induced nuclear factor-kappaB activation and I-kappaBalpha degradation: a fundamental mechanism for epidermal growth factor protection of the microtubule cytoskeleton and intestinal barrier integrity. AB - Oxidant damage and gut barrier disruption contribute to the pathogenesis of a variety of inflammatory gastrointestinal disorders, including inflammatory bowel disease (IBD). In our studies using a model of the gastrointestinal (GI) epithelial barrier, monolayers of intestinal (Caco-2) cells, we investigated damage to and protection of the monolayer barrier. We reported that activation of nuclear factor-kappaB (NF-kappaB) via degradation of its endogenous inhibitor I kappaBalpha is key to oxidant-induced disruption of barrier integrity and that growth factor (epidermal growth factor, EGF) protects against this injury by stabilizing the cytoskeletal filaments. Protein kinase C (PKC) activation seems to be required for monolayer maintenance, especially activation of the atypical zeta isoform of PKC. In an attempt to investigate, at the molecular level, the fundamental events underlying EGF protection against oxidant disruption, we tested the intriguing hypothesis that EGF-induced activation of PKC-zeta prevents oxidant-induced activation of NF-kappaB and the consequences of NF-kappaB activation, namely, cytoskeletal and barrier disruption. Monolayers of wild-type (WT) Caco-2 cells were incubated with oxidant (H2O2) with or without EGF or modulators. In other studies, we used the first gastrointestinal cell clones created by stable transfection of varying levels (1-5 microg) of cDNA to either overexpress PKC-zeta or to inhibit its expression. Transfected cell clones were then pretreated with EGF or a PKC activator (diacylglycerol analog 1-oleoyl-2 acetyl-glycerol, OAG) before oxidant. We monitored the following endpoints: monolayer barrier integrity, stability of the microtubule cytoskeleton, subcellular distribution and activity of the PKC-zeta isoform, intracellular levels and phosphorylation of the NF-kappaB inhibitor I-kappaBalpha, and nuclear translocation and activity of NF-kappaB subunits p65 and p50. Monolayers were also fractionated and processed to assess alterations in the structural protein of the microtubules, polymerized tubulin (S2), and monomeric tubulin (S1). Our data indicated that relative to WT monolayers exposed only to oxidant, pretreatment with EGF protected cell monolayers by 1) increasing native PKC-zeta activity; 2) decreasing several variables related to NF-kappaB activation [NF kappaB (both p50 and p65 subunits) nuclear translocation, NF-kappaB subunits activity, I-kappaBalpha degradation, and phosphorylation]; 3) increasing stable tubulin (increased polymerized S2 tubulin and decreased monomeric S1 tubulin); 4) maintaining the cytoarchitectural integrity of microtubules; and 5) preventing hyperpermeability (barrier disruption). In addition, relative to WT cells exposed to oxidant, monolayers of transfected cells stably overexpressing PKC-zeta (approximately 3.0-fold increase) were protected as indicated by decreases in all measures of NF-kappaB activation as well as enhanced stability of microtubule cytoarchitecture and barrier function. Overexpression induced stabilization of I kappaBalpha and inactivation of NF-kappaB was OAG-independent, although EGF potentiated this protection. Approximately 90% of the overexpressed PKC-zeta resided in particulate (membrane + cytoskeletal) fractions (with less than 10% in cytosolic fractions), indicating constitutive activation of the zeta isoform of PKC. Furthermore, antisense transfection to stably inhibit native PKC-zeta expression (-95%) and activation (-99%) prevented all measures of EGF-induced protection against NF-kappaB activation and monolayer disruption. We conclude the following: 1) EGF protects against oxidant disruption of the intestinal barrier integrity, in large part, through the activation of PKC-zeta and inactivation of NF-kappaB (an inflammatory mediator); 2) activation of PKC-zeta is by itself required for monolayer protection against oxidant stress of NF-kappaB activation; 3) the mechanism underlying this novel biological effect of the atypical PKC isoform zeta seems to involve suppression of phosphorylation and enhancement of stabilization of I-kappaBalpha; and 4) development of agents that can mimic or enhance PKC-zeta-induced suppression of NF-kappaB activation may be a useful therapeutic strategy for preventing oxidant damage to GI mucosal epithelium in disorders such as IBD. To our knowledge, this is the first report that PKC-zeta can inhibit the dynamics of NF-kappaB and cytoskeletal disassembly in cells. PMID- 12893840 TI - Chronic elevation of brain-derived neurotrophic factor by ampakines. AB - The ampakine CX614 positively modulates alpha-amino-3-hydroxy-5methyl-4 isoxazolepropionic acid (AMPA) receptor-gated currents and increases brain derived neurotrophic factor (BDNF) expression. In rat hippocampal slice cultures, CX614 rapidly increases BDNF gene expression but with time, mRNA levels fall despite the continued presence of active drug. The present study examined this apparent refractory period and the possibility that spaced ampakine treatments could sustain elevated BDNF protein levels. In cultured hippocampal slices, CX614, a second ampakine CX546, and the cholinergic agonist carbachol each increased BDNF mRNA levels with acute (3-h) treatment. After 4-day pretreatment with CX614, fresh ampakine (CX614 or CX546) did not induce BDNF mRNA, whereas carbachol did. Western blots confirmed that after an extended period of ampakine treatment, AMPA receptor protein levels are indeed reduced, suggesting that with longer treatments receptor down-regulation mediates ampakine insensitivity. Finally, using a "24-h on/24-h off" CX614 treatment protocol, the ampakine refractory state was circumvented, BDNF mRNA was induced with each ampakine application, and elevated BDNF protein levels were maintained through 5 days in vitro. These results suggest that spaced ampakine treatments can be used to sustain elevated neurotrophin levels and to test the utility of this manipulation for neuroprotection by endogenous neurotrophins. PMID- 12893841 TI - Potency and specificity of the pharmacological action of a new, antiasthmatic, topically administered soft steroid, etiprednol dicloacetate (BNP-166). AB - In the present study, the pharmacological effects of etiprednol dicloacetate (BNP 166; ethyl-17alpha-dichloroacetoxy-11beta-hydroxyandrosta-1,4-diene-3-one-17beta carboxylate), a new soft steroid, intended to use for the treatment of asthma, were investigated in an animal model of allergen sensitized and challenged Brown Norway rats using local treatment. The examinations involved the determination of the effect of the compound on the extent of allergen induced broncho-alveolar fluid and lung tissue eosinophilia, goblet cell hyperplasia and mucus production, perivascular edema formation, and airways hyperresponsiveness. The activity of etiprednol dicloacetate was compared with that of budesonide. Using in vitro methods, the soft character of etiprednol dicloacetate was investigated together with its capability to dissociate transrepressing and transactivating properties. We found that combining all the examined parameters etiprednol dicloacetate was at least equipotent with budesonide in the animal model, but in several investigated variables it surpassed the activity of budesonide. The effect of etiprednol dicloacetate in vitro was shown to be the function of the quantity of the serum, present in the assay, it was also strongly affected by the incubation time and decreased significantly when it was preincubated with human plasma. These features are characteristics of a soft drug that is quickly inactivated in the systemic circulation. In addition, it was revealed that while the transrepressing potential of etiprednol dicloacetate remained high, its transactivating activity was greatly reduced. These data indicate that the strong local effect of the compound will very likely be accompanied with a significantly reduced systemic activity predicting favorable selectivity in the pharmacological action of etiprednol dicloacetate. PMID- 12893842 TI - Metabolism of flavonoids via enteric recycling: mechanistic studies of disposition of apigenin in the Caco-2 cell culture model. AB - The purpose of this study was to determine the mechanisms responsible for intestinal disposition of apigenin in the human Caco-2 cell culture model. The results indicated that most of the absorbed apigenin (10 microM) were conjugated and only a small fraction was transported intact. The amounts of conjugates excreted, especially that of the sulfate, were dependent on days-post-seeding. Apical efflux of apigenin sulfate did not change with concentration of apigenin (4 to 40 microM), whereas its basolateral efflux increased (p < 0.01) with concentration and plateaued at about 25 microM. In contrast, sulfate formation rates in cell lysate increased with concentration and plateaued at 25 microM and were 4 to 6 times faster than the corresponding excretion rates. Formation and polarized excretion rates of glucuronidated apigenin increased with apigenin concentration but formation rates were usually 2.5 to 6 times faster than the corresponding excretion rates. Inhibitors of multidrug resistance-related proteins (MRPs) such as leukotriene C4 and MK-571, which inhibited glucuronidation of apigenin at a high concentration (>or=25 microM), significantly decreased excretion of both apigenin conjugates, and higher concentrations of MK-571 increased the extent of inhibition. In contrast, an organic anion transporter (OAT) inhibitor estrone sulfate only inhibited excretion of apigenin sulfate. In conclusion, we have shown for the first time that intestinal efflux is the rate-limiting step in the intestinal excretion of phase II conjugates of flavones. Furthermore, MRP and OAT are involved in the intestinal efflux of these hydrophilic phase II conjugates. PMID- 12893843 TI - Cholinesterase reactivation in vivo with a novel bis-oxime optimized by computer aided design. AB - Recently, several bis-pyridiniumaldoximes linked by a variable-length alkylene chain were rationally designed in our laboratories as cholinesterase reactivators. Extensive in vitro tests of these oximes with acetylcholinesterase inhibited by two different organophosphate agents, echothiophate and diisopropylfluorophosphate, revealed one compound with particularly good reactivation kinetics and affinity for phosphorylated acetylcholinesterase (AChE). This compound, designated "ortho-7", with a heptylene chain bridging two aldoximes ortho to a pyridinium ring nitrogen, was chosen for detailed comparison with the classic reactivator pyridine-2-aldoxime methochloride (2-PAM). In vitro, ortho-7 reactivated AChE selectively, without restoring activity of the related enzyme butyrylcholinesterase (BChE). For in vivo studies, rats were injected with ortho-7 or 2-PAM before or after organophosphate exposure, and the activities of AChE and BChE were determined at multiple intervals in blood and solid tissues. Ortho-7 behaved nearly as well in the animal as in vitro, reactivating AChE to the same extent as 2-PAM in all peripheral tissues studied (serum, red blood cell, and diaphragm), but at doses up to 100-fold smaller. Like other oxime reactivators, ortho-7 did not reactivate brain AChE after systemic administration. Nonetheless, this agent could be useful in combination therapy for organophosphate exposure, and it may provide a platform for development of additional, even more effective reactivators. PMID- 12893844 TI - Ethanol influences on native T-type calcium current in thalamic sleep circuitry. AB - Ethanol is known to disrupt normal sleep rhythms; however, the cellular basis for this influence is unknown. This study uses an in vitro slice preparation coupled with electrophysiological recordings to probe neuronal responses to acute ethanol exposure. Recordings were conducted in ferret and rat thalamic slices, since thalamic circuitry is an integral component of sleep/wake cycles and sleep spindles. A key mediator of spindle wave activity is the low-threshold calcium current (T-type current). The T-type current underlies burst responses in the lateral geniculate and thalamic reticular nuclei that are important in spindle propagation. Whole cell patch recordings in thalamic brain slices revealed that ethanol has a differential, dose-dependent effect on the native T-type current in thalamic relay cells. Low concentrations of ethanol (2.5, 5, and 10 mM) enhance T type current (n = 35), whereas higher concentrations of ethanol (20 and 50 mM) decrease T-type current (n = 27). To address whether this dose-dependent effect was due to variation between cells, in a subset we verified the differential effect within the same cell (n = 7). In an effort to examine whether the biphasic effects on the current were due to the order of ethanol exposures, we varied the order of high and low ethanol concentrations within the same cell. The ability of ethanol to perturb calcium currents in thalamic relay cells may provide a mechanistic framework for the well documented disruptions in sleep/wake behavior in subjects with ethanol exposure. PMID- 12893845 TI - Effects of intravenous cocaine and cigarette smoking on luteinizing hormone, testosterone, and prolactin in men. AB - Cocaine and nicotine have a number of similar behavioral and neurobiological effects. This study compared the acute effects of cocaine and cigarette smoking on luteinizing hormone (LH), testosterone (T), and prolactin. Twenty-four men who met American Psychiatric Association Diagnostic and Statistical Manual criteria for cocaine abuse or nicotine dependence were given intravenous cocaine (0.4 mg/kg) or placebo-cocaine, or smoked a low or high nicotine cigarette under controlled conditions. Placebo-cocaine or low nicotine cigarette smoking did not change LH, T, or prolactin. Peak plasma levels of 254 +/- 18 ng cocaine/ml and 22.6 +/- 3.4 ng nicotine/ml were measured at 8 and 14 min, respectively. LH increased significantly after both i.v. cocaine and high nicotine cigarette smoking (P < 0.01). These LH increases were significantly correlated with increases in cocaine and nicotine plasma levels (P < 0.001-0.003), and high nicotine cigarette smoking stimulated significantly greater increases in LH release than i.v. cocaine (P < 0.05). Testosterone levels did not change significantly after either cocaine or after high nicotine cigarette smoking. After i.v. cocaine, prolactin decreased significantly and remained below baseline levels throughout the sampling period (P < 0.05-0.01). After high nicotine cigarette smoking, prolactin increased to hyperpro-lactinemic levels within 6 min and remained significantly above baseline levels for 42 min (P < 0.05-0.03). The rapid increases in LH and reports of subjective "high" after both i.v. cocaine and high nicotine cigarette smoking illustrate the similarities between these drugs and suggest a possible contribution of LH to their abuse-related effects. PMID- 12893846 TI - A natural product ligand of the oxysterol receptor, liver X receptor. AB - Natural products have been identified as ligands for a number of members of the nuclear hormone receptor (NHR) superfamily. Often these natural products are used as dietary supplements to treat myriad ailments ranging from perimenopausal hot flashes to hypercholesterolemia and reduced cognitive function. Examples of some natural product ligands for NHRs include genestein (estrogen receptors NR3A1 and NR3A2), guggulsterone (farnesoid X receptor NR1H4), and St. John's wort (pregnane X receptor, NR1I2). In this study, we identified the first nonoxysterol natural product that functions as a ligand for the liver X receptor (LXRalpha and LXRbeta; NR1H3, NR1H2), a NHR that acts as the receptor for oxysterols and plays a key role in regulation of cholesterol metabolism and transport as well as glucose metabolism. We show that paxilline, a fungal metabolite, is an efficacious agonist of both LXRalpha and LXRbeta in biochemical and in vitro cell based assays. Paxilline binds directly to both receptors and is an activator of LXR-dependent transcription in cell-based reporter assays. We also demonstrate that paxilline binding to the receptors results in efficient activation of transcription of two physiological LXR target genes, ABCA1 and SREBP. The discovery of paxilline, the first reported nonoxysterol natural product ligand of the LXRs, may provide insight into the mechanism of ligand recognition by these receptors and reaffirms the utility of examining natural product libraries for identifying novel NHR ligands. PMID- 12893847 TI - Crucial role of histamine for regulation of gastric acid secretion ascertained by histidine decarboxylase-knockout mice. AB - Histidine decarboxylase (HDC) represents the sole enzyme that produces histamine in the body. The present work investigated the role of endogenous histamine in carbachol- and gastrin-induced gastric acid secretion with HDC-knockout (HDC-/-) mice. Acid secretion was measured in either mice subjected to acute fistula production under urethane anesthesia or conscious mice that had previously undergone pylorus ligation. In wild-type mice, carbachol and gastrin significantly stimulated acid secretion, increasing gastric mucosal histamine. In contrast, in HDC-/- mice, carbachol and gastrin had little impact when either delivered alone or together. Nonetheless, the two agents achieved a synergistic effect when delivered together with exogenous histamine, stimulating acid secretion in HDC-/- mice. Such synergism was abolished by the histamine H2 receptor antagonist famotidine. cAMP involvement in acid secretion was also examined with theophylline, a phosphodiesterase inhibitor, and forskolin, an adenylate cyclase activator. In wild-type mice, theophylline significantly increased acid secretion, enhancing carbachol- and gastrin-stimulated acid secretion. In contrast, in HDC-/- mice, theophylline failed to exert an effect on basal acid secretion, as well as carbachol- and gastrin-stimulated acid secretion. Although forskolin interacted with carbachol, allowing acid secretion in HDC-/- mice, similar results were not achieved with gastrin. Such results suggest that 1) histamine is essential for carbachol- and gastrin-stimulated gastric acid secretion in mice; and 2) histamine-induced cAMP production contributes to the in vivo response to carbachol or gastrin. PMID- 12893848 TI - Antisense oligonucleotide inhibition of Bcl-xL and Bid expression in liver regulates responses in a mouse model of Fas-induced fulminant hepatitis. AB - Activation of the cell-surface receptor Fas can lead to apoptosis in parenchymal cells in the liver, and if severe enough, result in fulminant hepatic failure and animal death. In the present study, we have examined the roles played by the Bcl 2 family members Bcl-xL and Bid in regulating this response. To do this, we have developed chemically modified 2'-O-(2-methoxy) ethyl antisense inhibitors of both Bid and Bcl-xL expression. In Balb/c mice, dosing with these antisense oligonucleotides reduced expression of the targeted mRNA by greater than 80% in the liver. This reduction was highly dependent upon oligonucleotide sequence and oligonucleotide dose. Reduction of Bcl-xL expression resulted in a potentiation of Fas-mediated apoptosis in liver and significant increase of the lethality of Fas-mediated fulminant hepatitis (p < 0.0001). In contrast, reduction of Bid expression protected the animals against Fas-mediated fulminant hepatitis and death (p < 0.0001). Simultaneous dosing of mice with Bcl-xL and Bid-targeting antisense oligonucleotides resulted in an inhibition of expression of both targeted proteins and protection of the animals from Fas-mediated apoptosis. These results demonstrate, for the first time, the role of Bcl-xL in regulating responses to proapoptotic Fas signaling in mouse liver. In addition, this is the first reported example demonstrating the ability of antisense inhibitors to reduce expression of multiple proteins in animals by simultaneous dosing. PMID- 12893849 TI - Type 2 diabetes is cardiovascular disease. PMID- 12893850 TI - Rationing treatment on the NHS--still a political issue. PMID- 12893851 TI - The SARS epidemic in Hong Kong: what lessons have we learned? PMID- 12893852 TI - Identifying neuropathic pain in patients with head and neck cancer: use of the Leeds Assessment of Neuropathic Symptoms and Signs Scale. AB - The Leeds Assessment of Neuropathic Symptoms and Signs Scale (LANSS) is a simple bedside test in two parts-a patient-completed questionnaire and a brief clinical assessment. Its diagnostic capabilities have never been tested in patients with cancer pain. To determine these we conducted a prospective study in outpatients with head and neck cancer. All patients with pain completed the LANSS and underwent a medical assessment with a palliative care physician, whose findings were then reviewed by a pain specialist blinded to the LANSS scores. We assessed acceptability and understanding of the LANSS by patients and calculated the sensitivity and specificity of total LANSS scores and subscores derived from the patient-completed section. Of 130 patients approached, 125 took part. 25 (20%) of these had cancer related pain, mean score on an 11 point numerical rating scale 6.3 (median 6.0, range 3-10). Average age was 60 years (median 60, range 27-84); 56% were male. LANSS completion time was about five minutes, and the procedure was acceptable to all patients. The pain specialist diagnosed neuropathic pain in 14/25 patients, in 13 of whom the neuropathic pain was part of a mixed pain picture. The LANSS correctly identified 11 of these cases (sensitivity 79%; specificity 100%). The patient-completed section alone had a sensitivity of 86% and a specificity of 91%. The LANSS is a simple and suitable screening test for neuropathic pain in patients with head and neck cancer related pain, although some modifications might improve it. PMID- 12893853 TI - Association between course of study at university and cause-specific mortality. AB - Although socioeconomic position is clearly related to mortality and one measure of this is length of education, it is not known whether the choice of course at university determines future health. We therefore investigated the association between faculty of study and all-cause and cause-specific mortality in a prospective follow-up of male students who underwent health examinations while attending Glasgow University from 1948 to 1968. Among the 9887 (84%) alumni traced by means of the NHS Central Register, 8367 (85%) had full data on important potential confounding variables; 939 of these men had died. Physiological variables differed little between students from the various faculties. Medical students were most likely to come from affluent social backgrounds and, after law students, were most likely to be smokers. Compared with former medical students, former arts and law students had excess all-cause and cardiovascular disease mortality, while science and engineering alumni had similar risks. Former medical students had lower lung cancer mortality than other alumni but higher mortality from alcohol-related causes including accidents, suicide and violence. The lower mortality risks observed among former medical and engineering students may be due to their better employment prospects and healthier lifestyle behaviours, although the high mortality from alcohol-related causes among former medical students underscores the complexity of choice of health behaviour. The findings point to the potential for disease prevention among the large proportion of the population who now have third-level education. PMID- 12893854 TI - Vaccination strategies to reduce the risk of leukaemia and melanoma. PMID- 12893855 TI - Diagnosis of perforated enterocystoplasty. AB - In the operation of enterocystoplasty, now widely practised, segments of bowel are used to augment or replace the urinary bladder. An occasional complication is perforation, and this may present in non-specialist settings. We investigated the management of spontaneous perforations among 264 patients with enterocystoplasty followed by one surgeon for 2-18 years. Patients' charts were examined for data on presentation, diagnosis and treatment. 10 patients had thirteen perforations; data were available for nine perforations in 9 patients. Mean time from enterocystoplasty to perforation was 45 months. Presentation was shoulder pain in 1 and abdominal pain (with or without fever) in 8. Perforation was diagnosed without delay in 3, but the initial diagnosis was urinary tract infection in 4 and small-bowel obstruction in 2. Ultrasound was the most useful investigation being diagnostic in 6 of 7 cases; contrast cystography showed a leak in only 2 of the 6 patients in whom it was performed. Treatment was successful in 8 cases (surgery 6; percutaneous drainage 2); 1 patient, who remained undiagnosed, was treated medically and died. Patients with enterocystoplasty need to be educated about this potentially lethal complication, so that they can alert non-specialist clinicians to what may have happened. In any patient with enterocystoplasty who reports abdominal pain or shoulder pain, perforation has to be ruled out. PMID- 12893856 TI - Use of information technology in general practice. AB - The internet and NHS Net are used increasingly in UK general practice. A questionnaire survey conducted in southern England examined these applications. 77 (55%) of 141 practices responded. Of these, 71 were connected to one or other service and 27 offered a practice website. Only a small minority used a website for direct patient booking or access to pathology results. Moreover, among those with a practice website, none paid the necessary attention to data security. The survey revealed some fundamental misunderstandings that may partly account for the slow uptake of these technologies in British general practice. PMID- 12893857 TI - How will the two-weeks-wait rule affect delays in management of urological cancers? AB - The UK National Health Service has now specified a maximum interval of two weeks between general practitioner (GP) referral and specialist assessment for patients with suspected cancer. We examined progress through the cancer pathway in 160 patients with potentially curable cancers of the prostate, bladder, kidney and testis before implementation of this rule. Median intervals with interquartile ranges were quantified from the first GP consultation to hospital referral, then to the first hospital consultation, confirmation of diagnosis and definitive surgery. 34% of patients were seen at the hospital within two weeks of referral. The overall median interval from GP consultation to radical surgery was 137 days, the longest being for prostate cancer (median 244). For prostate, bladder and renal cancers the principal element of delay was from the time of diagnosis to surgery (76, 73 and 26 days respectively). These results indicate that, under the two-weeks-wait rule, 2 out of every 3 patients achieve earlier initial assessment. However, the overall delay will not be substantially reduced without concomitant increases in diagnostic facilities, theatre time and human resources. PMID- 12893858 TI - Totally endoscopic robotically enhanced coronary artery bypass on the beating heart. PMID- 12893859 TI - Swollen lips and a tender neck. PMID- 12893860 TI - Pathophysiological insights from a case of reversible pulmonary arterial hypertension. PMID- 12893861 TI - Lymphadenopathy after joint replacement for osteoclastoma. PMID- 12893862 TI - Recurrent bowel infarction in paroxysmal nocturnal haemoglobinuria. PMID- 12893863 TI - Priapism at age 94. PMID- 12893864 TI - Mammographic screening: give us the facts. PMID- 12893865 TI - A new kind of living will. PMID- 12893866 TI - Cerebrovascular disease in Dumas' The count of Monte Cristo. PMID- 12893867 TI - Medically unexplained symptoms. PMID- 12893868 TI - Medically unexplained symptoms. PMID- 12893869 TI - Effects of exposure to CS. PMID- 12893870 TI - Abdominal pregnancy and lithopaedion. PMID- 12893871 TI - Maurice Ravel's amusia. PMID- 12893872 TI - What about GPs in the fight against tuberculosis? PMID- 12893873 TI - The sex peptide of Drosophila melanogaster: female post-mating responses analyzed by using RNA interference. AB - Mating induces profound changes in female insect behavior and physiology. In Drosophila melanogaster, mating causes a reduction in sexual receptivity and an elevation in egg production for at least 5 days. Injection of the seminal fluid sex peptide (SP) induces both responses in virgin females, but only for 1-2 days. The role of SP in eliciting the responses to mating remains to be elucidated. Functional redundancy between seminal fluid components may occur. In addition, mating with spermless males results in brief (1- to 2-day) post-mating responses, indicating either that there is a "sperm effect" or that sperm act as carriers for SP or other seminal fluid components. Here we used RNA interference to suppress SP expression, to determine whether SP is required to elicit full post mating responses, the magnitude of responses due to other seminal fluid components, and whether SP accounts for the "sperm effect." Receptivity was higher and egg production lower in females mated to SP knock-down males than in controls. Comparison with virgins showed that the responses were brief. SP is therefore required for normal magnitude and persistence of postmating responses. Sperm transfer and use were normal in mates of SP knock-down males, yet their post-mating responses were briefer than after normal matings, and similar to those reported in mates of spermless son-of-tudor males. The prolonged "sperm effect" on female receptivity and egg production is therefore entirely attributable to SP, but sperm are necessary for its occurrence. PMID- 12893875 TI - Combinatorial auction design. AB - Combinatorial auctions allow for more expressive bidding in which participants can submit package bids with logical constraints that limit allowable outcomes. This type of auction can be useful when participants' values are complementary or when participants have production and financial constraints. However, combinatorial auctions are currently rare in practice. The main problems confronted in implementing these auctions are that they have computational uncertainty (i.e., there is no guarantee that the winning bids for such an auction can be found in a "reasonable" amount of time when the number of bidders and items becomes larger) and that the auction is cognitively complex and can lead participants to pursue perverse bidding strategies. This article describes a type of combinatorial auction that, during laboratory testing, eliminated these problems and produced extremely efficient outcomes. PMID- 12893874 TI - Four functionally distinct C-type natriuretic peptides found in fish reveal evolutionary history of the natriuretic peptide system. AB - Natriuretic peptides (NPs) are major cardiovascular and osmoregulatory hormones in vertebrates. Although tetrapods generally have three subtypes, atrial NP (ANP), B-type NP (BNP), and C-type NP (CNP), some teleosts lack BNP, and sharks and hagfish have only one NP. Thus, NPs have diverged during fish evolution, possibly reflecting changes in osmoregulatory systems. In this study, we found, by cDNA cloning, four distinct CNPs (1 through 4) in the medaka (Oryzias latipes) and puffer fish (Takifugu rubripes), although to our knowledge no more than two CNPs have been isolated from a vertebrate species. Predicted mature CNP-1 was most similar, and CNP-4 was most dissimilar, to mammalian CNPs. However, synthetic CNP-4 most potently activated OlGC1, a medaka CNP-specific receptor (NPR-B) expressed in cultured cells, whereas CNP-1 and CNP-3 most activated OlGC7 and OlGC2, two medaka homologues of the ANP/BNP receptor (NPR-A), respectively. Linkage mapping in medaka followed by comparative genomic analyses among fishes and humans located four CNP genes in separate medaka chromosomes corresponding to human chromosomes 1, 2, 12, and 17. From conserved synteny, the following evolutionary history of NPs was evoked: (i) four CNP genes were generated by chromosomal duplications before the divergence of elasmobranchs; (ii) the CNP-3 gene generated ANP and BNP genes through tandem duplication before the divergence of tetrapods and teleosts; (iii) CNP-1 and -2 genes were retained in the teleost lineage but not in the tetrapod lineage; (iv) the CNP-3 gene disappeared from the tetrapod lineage after divergence of amphibians; and (v) the CNP-4 gene is retained in humans as CNP. PMID- 12893876 TI - Advantages of external periodic events to the evolution of biochemical oscillatory reactions. AB - We compare, by calculations on a simple model of glycolysis, the evolutionary development of oscillatory reaction mechanisms in the presence and absence of external periodic events, such as an oscillatory or constant influx of glucose in an open reaction system. The chosen model has autonomous oscillations for given choices of the parameters of the feedback loops responsible for the oscillations, and for a given range of the total adenylate pool concentration. We change first one, then two of the parameters, so that there are no autonomous oscillations, and then vary these parameters with a genetic algorithm method in which the parameters are represented by binary strings that evolve by selection, crossover, and mutations; the optimization goal is the attainment of a high ATP/ADP concentration ratio in the system. This goal is taken to provide evolutionary advantages and is shown to be achieved more quickly in the presence of external periodic events, rather than constant influx of glucose. The results suggest the possibility of the enhanced evolutionary development of oscillatory biological reactions at shores where waves impinge on rocks and bring nutrients periodically. Measurements have shown that animals and plants grow more rapidly in the presence of such wave action than in its absence. PMID- 12893877 TI - Traveling stripes on the skin of a mutant mouse. AB - In the course of animal development, complex structures form autonomously from the apparently shapeless egg. How cells can produce spatial patterns that are much larger than each cell is one of the key issues in developmental biology. It has been suggested that spatial patterns in animals form through the same principles by which dispatched structures are formed in the nonbiological system. However, because of the complexity of biological systems, molecular details of such phenomena have been rarely clarified. In this article, we introduce an example of a pattern-forming phenomenon that occurs in the skin of mutant mice. The mutant mouse has a defect in splicing of the Foxn1 (Whn or nude) gene, which terminates hair follicle development just after pigment begins to accumulate in the follicle. The immature follicles are rapidly discharged, and a new hair cycle resumes. Eventually, the skin color of the mouse appears to oscillate. The color oscillation is synchronous in juvenile mice, but the phase gradually shifts among skin regions to eventually form traveling, evenly spaced stripes. Although the time scale is quite different, the pattern change in the mutant mouse shares characteristics with the nonlinear waves generated on excitable media, such as the Belousov-Zhabotinskii reaction, suggesting that a common principle underlies the wave pattern formation. Molecular details that underlie the phenomenon can be conjectured from recent molecular studies. PMID- 12893878 TI - Beta1 subunits facilitate gating of BK channels by acting through the Ca2+, but not the Mg2+, activating mechanisms. AB - The beta1 subunit of BK (large conductance Ca2+ and voltage-activated K+) channels is essential for many key physiological processes, such as controlling the contraction of smooth muscle and the tuning of hair cells in the cochlea. Although it is known that the beta1 subunit greatly increases the open probability of BK channels, little is known about its mechanism of action. We now explore this mechanism by using channels in which the Ca2+- and Mg2+-dependent activating mechanisms have been disrupted by mutating three sites to remove the Ca2+ and Mg2+ sensitivity. We find that the presence of the beta1 subunit partially restores Ca2+ sensitivity to the triply mutated channels, but not the Mg2+ sensitivity. We also find that the beta1 subunit has no effect on the Mg2+ sensitivity of WT BK channels, in contrast to its pronounced effect of increasing the apparent Ca2+ sensitivity. These observations suggest that the beta1 subunit increases open probability by working through the Ca2+-dependent, rather than Mg2+-dependent, activating mechanisms, and that the action of the beta1 subunit is not directly on the Ca2+ binding sites, but on the allosteric machinery coupling the sites to the gate. The differential effects of the beta1 subunit on the Ca2+ and Mg2+ activation of the channel suggest that these processes act separately. Finally, we show that Mgi2+ inhibits, rather than activates, BK channels in the presence of the beta1 subunit for intermediate levels of Cai2+. This Mg2+ inhibition in the presence of the beta1 subunit provides an additional regulatory mechanism of BK channel activity. PMID- 12893879 TI - A SNARE required for retrograde transport to the endoplasmic reticulum. AB - SNAREs (soluble N-ethylmaleimide-sensitive factor attachment protein receptors) are central components of the machinery mediating membrane fusion in all eukaryotic cells. Sequence analysis of the yeast genome revealed a previously uncharacterized SNARE, SNARE-like tail-anchored protein 1 (Slt1). Slt1 is an essential protein localized in the endoplasmic reticulum (ER). It forms a SNARE complex with Sec22 and the ER syntaxin Ufe1. Down-regulation of Slt1 levels leads to improper secretion of proteins normally resident in the ER. We suggest that Slt1 is a component of the SNAREpin required for retrograde traffic to the ER. Based on the previously reported association with Ufe1 and Sec22, Sec20 likely contributes the fourth SNARE to the SNAREpin. PMID- 12893880 TI - Activation functions 1 and 2 of nuclear receptors: molecular strategies for transcriptional activation. AB - Nuclear receptors (NRs) comprise a family of ligand inducible transcription factors. To achieve transcriptional activation of target genes, DNA-bound NRs directly recruit general transcription factors (GTFs) to the preinitiation complex or bind intermediary factors, so-called coactivators. These coactivators often constitute subunits of larger multiprotein complexes that act at several functional levels, such as chromatin remodeling, enzymatic modification of histone tails, or modulation of the preinitiation complex via interactions with RNA polymerase II and GTFs. The binding of NR to coactivators is often mediated through one of its activation domains. Many NRs have at least two activation domains, the ligand-independent activation function (AF)-1, which resides in the N-terminal domain, and the ligand-dependent AF-2, which is localized in the C terminal domain. In this review, we summarize and discuss current knowledge regarding the molecular mechanisms of AF-1- and AF-2-mediated gene activation, focusing on AF-1 and AF-2 conformation and coactivator binding. PMID- 12893881 TI - Two distinct coactivators, DRIP/mediator and SRC/p160, are differentially involved in vitamin D receptor transactivation during keratinocyte differentiation. AB - Cell programs such as proliferation and differentiation involve the sequential activation and repression of gene expression. Vitamin D, via its active metabolite 1,25-dihydroxyvitamin D [1,25-(OH)2D3)], controls the proliferation and differentiation of a number of cell types, including keratinocytes, by directly regulating transcription. Two classes of coactivators, the vitamin D receptor (VDR)-interacting proteins (DRIP/mediator) and the p160 steroid receptor coactivator family (SRC/p160), control the actions of nuclear hormone receptors, including the VDR. However, the relationship between these two classes of coactivators is not clear. Using glutathione-S-transferase-VDR affinity beads, we have identified the DRIP/mediator complex as the major VDR binding complex in proliferating keratinocytes. After the cells differentiated, members of the SRC/p160 family were identified in the complex but not major DRIP subunits. Both DRIP and SRC proteins were expressed in keratinocytes. DRIP205 expression decreased during differentiation, although SRC-3 levels increased. Both DRIP205 and SRC-3 potentiated vitamin D-induced transcription in proliferating cells, but during differentiation, DRIP205 was no longer effective. These results indicate that these two distinct coactivators are sequentially involved in vitamin D regulation of gene transcription during keratinocyte differentiation, suggesting that these coactivators are part of the means by which the temporal sequence of gene expression is regulated during the differentiation process. PMID- 12893882 TI - Estrogen receptor-dependent genomic responses in the uterus mirror the biphasic physiological response to estrogen. AB - The physiological responses of the rodent uterus to acute estrogen (E) dosing can be divided into early and late events. Examples of early responses include increased RNA transcription, hyperemia, and water imbibition 2 and 6 h following E administration respectively, whereas later responses include cycles of DNA synthesis and mitosis of epithelial cells beginning 10 and 16 h after E. The development of estrogen receptor (ER) knockout (ERKO) mice, combined with microarray technology, has allowed us to design a genomic approach to study the acute response of the rodent reproductive tract to E. To determine whether early and late biological responses are correlated with altered regulation of a single set of genes or distinct sets of genes characteristic of early and late responses, uterine RNA was obtained from ovariectomized mice that were treated with vehicle or with estradiol for 2 h (early) or 24 h (late). Samples were also prepared from identically treated mice that lacked either ERalpha (alphaERKO) or ERbeta (betaERKO) to address the relative contributions of the ERs in the uterine responses. Microarray analysis of the relative expression of 8700 mouse cDNAs indicated distinct clusters of genes that were regulated both positively and negatively by E in the early or late phases as well as clusters of genes regulated at both times. Both early and late responses by the betaERKO samples were indistinguishable from those of WT samples, whereas the alphaERKO showed little change in gene expression in response to E, indicating the predominant role for ERalpha in the genomic response. Further studies indicated that the genomic responses in samples from intermediate time points (6 h, 12 h) fall within the early or late clusters, rather than showing unique clusters regulated in the intermediary period. The use of this genomic approach has illustrated how physiological responses are reflected in genomic patterns. Furthermore, the identification of functional gene families that are regulated by E in the uterus combined with the utilization of genetically altered experimental animal models can help to uncover and define novel mechanisms of E action. PMID- 12893883 TI - Retinoid X receptor is a nonsilent major contributor to vitamin D receptor mediated transcriptional activation. AB - The vitamin D receptor (VDR) belongs to the thyroid hormone/retinoid receptor subfamily of nuclear receptors and functions as a heterodimer with retinoid X receptor (RXR). The RXR-VDR heterodimer, in contrast to other members of the class II nuclear receptor subfamily, is nonpermissive where RXR does not bind its cognate ligand, and therefore its role in VDR-mediated transactivation by liganded RXR-VDR has not been fully characterized. Here, we show a unique facet of the intermolecular RXR-VDR interaction, in which RXR actively participates in vitamin D3-dependent gene transcription. Using helix 3 and helix 12 mutants of VDR and RXR, we provide functional evidence that liganded VDR allosterically modifies RXR from an apo (unliganded)- to a holo (liganded)-receptor conformation, in the absence of RXR ligand. As a result of the proposed allosteric modification of RXR by liganded VDR, the heterodimerized RXR shows the "phantom ligand effect" and thus acquires the capability to recruit coactivators steroid receptor coactivator 1, transcriptional intermediary factor 2, and amplified in breast cancer-1. Finally, using a biochemical approach with purified proteins, we show that RXR augments the 1,25-dihydroxyvitamin D3-dependent recruitment of transcriptional intermediary factor 2 in the context of RXR-VDR heterodimer. These results confirm and extend the previous observations suggesting that RXR is a significant contributor to VDR-mediated gene expression and provide a mechanism by which RXR acts as a major contributor to vitamin D3 dependent transcription. PMID- 12893884 TI - Progesterone regulation of the mammalian ortholog of methylcitrate dehydratase (immune response gene 1) in the uterine epithelium during implantation through the protein kinase C pathway. AB - Implantation requires coordination between development of the blastocyst and the sex steroid hormone-regulated differentiation of the uterus. Under the influence of these hormones, the uterine luminal epithelium becomes receptive to attachment of the hatched blastocyst. In this study we sought to identify genes regulated by progesterone (P4) in the uterine epithelium. This resulted in the identification of one novel P4-regulated gene that had been previously found in lipopolysaccharide-stimulated macrophages and called immune response gene-1 (Irg1) and which is the mammalian ortholog of the bacterial gene encoding methylcitrate dehydratase. In adult mice Irg1 expression was limited to the uterine luminal epithelium where it is expressed only during pregnancy with a peak coinciding with implantation. Irg1 mRNA expression is regulated synergistically by P4 and estradiol (E2) but not by E2 alone. In macrophages Irg1 is induced by lipopolysaccharide through a protein kinase C (PKC)-regulated pathway. Now we demonstrate that the PKC pathway is induced in the uterine epithelium at implantation by the synergistic action of P4 and E2 and is responsible for the hormone induction of Irg1. These results suggest that the PKC pathway plays an important role in modulating steroid hormone responsiveness in the uterine luminal epithelium during the implantation window and that Irg1 will be an important marker of this window and may play an important role in implantation. PMID- 12893885 TI - SIR1, an upstream component in auxin signaling identified by chemical genetics. AB - Auxin is a plant hormone that regulates many aspects of plant growth and development. We used a chemical genetics approach to identify SIR1, a regulator of many auxin-inducible genes. The sir1 mutant was resistant to sirtinol, a small molecule that activates many auxin-inducible genes and promotes auxin-related developmental phenotypes. SIR1 is predicted to encode a protein composed of a ubiquitin-activating enzyme E1-like domain and a Rhodanese-like domain homologous to that of prolyl isomerase. We suggest a molecular context for how the auxin signal is propagated to exert its biological effects. PMID- 12893886 TI - Coherent spin transfer between molecularly bridged quantum dots. AB - Femtosecond time-resolved Faraday rotation spectroscopy reveals the instantaneous transfer of spin coherence through conjugated molecular bridges spanning quantum dots of different size over a broad range of temperature. The room-temperature spin-transfer efficiency is approximately 20%, showing that conjugated molecules can be used not only as interconnections for the hierarchical assembly of functional networks but also as efficient spin channels. The results suggest that this class of structures may be useful as two-spin quantum devices operating at ambient temperatures and may offer promising opportunities for future versatile molecule-based spintronic technologies. PMID- 12893887 TI - Discovery of gene function by expression profiling of the malaria parasite life cycle. AB - The completion of the genome sequence for Plasmodium falciparum, the species responsible for most malaria human deaths, has the potential to reveal hundreds of new drug targets and proteins involved in pathogenesis. However, only approximately 35% of the genes code for proteins with an identifiable function. The absence of routine genetic tools for studying Plasmodium parasites suggests that this number is unlikely to change quickly if conventional serial methods are used to characterize encoded proteins. Here, we use a high-density oligonucleotide array to generate expression profiles of human and mosquito stages of the malaria parasite's life cycle. Genes with highly correlated levels and temporal patterns of expression were often involved in similar functions or cellular processes. PMID- 12893888 TI - A microRNA as a translational repressor of APETALA2 in Arabidopsis flower development. AB - Plant microRNAs (miRNAs) show a high degree of sequence complementarity to, and are believed to guide the cleavage of, their target messenger RNAs. Here, I show that miRNA172, which can base-pair with the messenger RNA of a floral homeotic gene, APETALA2, regulates APETALA2 expression primarily through translational inhibition. Elevated miRNA172 accumulation results in floral organ identity defects similar to those in loss-of-function apetala2 mutants. Elevated levels of mutant APETALA2 RNA with disrupted miRNA172 base pairing, but not wild-type APETALA2 RNA, result in elevated levels of APETALA2 protein and severe floral patterning defects. Therefore, miRNA172 likely acts in cell-fate specification as a translational repressor of APETALA2 in Arabidopsis flower development. PMID- 12893889 TI - Radiology 2003--science to practice. PMID- 12893890 TI - Science to practice: blood flow quantification with contrast-enhanced US: "entrance in the section" phenomenon--phantom and rabbit study. PMID- 12893892 TI - The end of life. PMID- 12893893 TI - Lymphoscintigraphy for lymphatic mapping in breast carcinoma. PMID- 12893894 TI - Opportunities and challenges for the National Institute of Biomedical Imaging and Bioengineering. PMID- 12893895 TI - Image-guided tumor ablation: proposal for standardization of terms and reporting criteria. AB - The field of image-guided tumor ablation requires standardization of terms and reporting criteria to facilitate effective communication of ideas and appropriate comparison between treatments with different technologies, such as chemical ablation (ethanol or acetic acid) and thermal therapies, such as radiofrequency, laser, microwave, ultrasound, and cryoablation. On the basis of this premise, a working committee was established with the goal of producing a proposal on such standardization. The intent of the Working Group is to provide a framework that will facilitate the clearest communication between investigators and will provide the greatest flexibility in comparisons between the many new, exciting, and emerging technologies. The members of the Working Group now propose a vehicle for reporting the various aspects of image-guided ablation therapy, including classifications of therapies and procedures, appropriate descriptors of image guidance, and terms to define imaging and pathologic findings. Methods for standardizing the reporting of follow-up findings and complications and other important aspects that require attention when reporting clinical results are addressed. It is the group's hope and intention that adherence to the recommendations of this proposal will facilitate achievement of the group's main objective: improved precision and communication in this field that lead to more accurate comparison of technologies and results and ultimately to improved patient outcomes. PMID- 12893896 TI - Self-referred whole-body CT imaging: current implications for health care consumers. AB - PURPOSE: To conduct an empirical analysis of self-referred whole-body computed tomography (CT) and develop a profile of the geographic and demographic distribution of centers, types of services and modalities, costs, and procedures for reporting results. MATERIALS AND METHODS: An analysis was conducted of Web sites for imaging centers accepting self-referred patients identified by two widely used Internet search engines with large indexes. These Web sites were analyzed for geographic location, type of screening center, services, costs, and procedures for managing imaging results. Demographic data were extrapolated for analysis on the basis of center location. Descriptive statistics, such as frequencies, means, SDs, ranges, and CIs, were generated to describe the characteristics of the samples. Data were compared with national norms by using a distribution-free method for calculating a 95% CI (P <.05) for the median. RESULTS: Eighty-eight centers identified with the search methods were widely distributed across the United States, with a concentration on both coasts. Demographic analysis further situated them in areas of the country characterized by a population that consisted largely of European Americans (P <.05) and individuals of higher education (P <.05) and socioeconomic status (P <.05). Forty seven centers offered whole-body screening; heart and lung examinations were most frequently offered. Procedures for reporting results were highly variable. CONCLUSION: The geographic distribution of the centers suggests target populations of educated health-conscious consumers who can assume high out-of pocket costs. Guidelines developed from within the profession and further research are needed to ensure that benefits of these services outweigh risks to individuals and the health care system. PMID- 12893897 TI - Dose reduction in pediatric CT: a rational approach. AB - PURPOSE: To determine size-dependent technique factors for pediatric computed tomography (CT) by using physically measured objective data. MATERIALS AND METHODS: Six phantom cylinders (10-32 cm in diameter) were scanned with a clinical multi-detector row CT scanner. CT noise was statistically characterized for CT technique factors from 80 to 140 kVp and from 10 to 300 mAs. Dose measurements were performed with each phantom. Dilute iodine and tissue contrast were determined with computer calculations validated with measured data. The dose, noise, and contrast data were computer fit, and pediatric CT technique factors (milliampere seconds) necessary to maintain the contrast-to-noise ratio (CNR) were computed. RESULTS: As compared with that in a reference cylindric adult abdomen of 28 cm in diameter, CNR was maintained at a constant level in pediatric patients of 25, 20, and 15 cm in diameter, respectively, when milliampere second values of 0.557, 0.196, and 0.054 of the adult milliampere second values were used. The corresponding doses were reduced to 0.642, 0.287, and 0.090 of the 28-cm-diameter adult dose, respectively. CT techniques for examination of pediatric heads measuring 15 and 13 cm, respectively, can involve the use of milliampere second values of 0.572 and 0.366 of those used for examination of a standard 17-cm-diameter adult head. CONCLUSION: CT technique charts for pediatric abdominal and head examinations were produced on the basis of physically measured data; use of these tables will enable pediatric radiation dose to be reduced while CNR is preserved. PMID- 12893898 TI - Intrapleural fibrinolysis for parapneumonic effusion and empyema in children. AB - PURPOSE: To assess the safety and efficacy of urokinase and alteplase for intrapleural fibrinolysis in children with parapneumonic pleural fluid collections. MATERIALS AND METHODS: A retrospective review was performed of 71 children with parapneumonic pleural fluid accumulations who were treated with thoracostomy tube placement and intrapleural instillation of either urokinase or alteplase. The procedures were performed with urokinase between September 2, 1995, and March 27, 1998, and with alteplase between March 30, 1998, and January 2, 2002. The medical records and daily chest radiographs were reviewed by a pediatric radiologist to ascertain demographic information, signs and symptoms, laboratory results, thoracostomy tube output, treatment details, and radiographic pleural thickness and lung opacification. Multiple variables were compared for the alteplase and urokinase groups by using univariate and multivariate statistics. We defined primary treatment success as resolution of signs and symptoms at the time of discharge, without surgical intervention. RESULTS: Primary treatment success was 98% for alteplase and 100% for urokinase, with no major complications. Greater pleural fluid drainage occurred with alteplase than urokinase during the 1st (P =.001) and 2nd (P =.002) days of fibrinolytic therapy, and for the duration of thoracostomy drainage (P <.001). Multivariate models showed greater total drainage with alteplase (P <.001), greater patient age (P <.001), larger tube size (P =.002), and greater volume of drainage during the 24 hours prior to fibrinolysis (P <.001). CONCLUSION: Intrapleural fibrinolysis with urokinase or alteplase facilitates thoracostomy tube drainage of parapneumonic pleural fluid. With the dosing regimen used in this study, alteplase produces greater thoracostomy tube output than does urokinase. PMID- 12893899 TI - The deep sulcus sign. PMID- 12893900 TI - Irreversible JPEG compression of digital chest radiographs for primary interpretation: assessment of visually lossless threshold. AB - PURPOSE: To determine if digital chest images could be compressed in a primary interpretation context without perceived loss of fidelity (below the visually lossless threshold) at transilluminated film or cathode ray tube (CRT) display. MATERIALS AND METHODS: One hundred forty-four posteroanterior radiographs were obtained with a digital chest radiography system. At both film and CRT display, an identified original image was presented side by side with a replicate, which was either an unaltered image or an image that had been Joint Photographic Experts Group (JPEG) compressed to 10:1, 20:1, or 50:1 and reconstructed. Each of the 10 readers indicated whether the replicate was "indistinguishable from the original" or "degraded" at clinical reading distance and at close inspection. The readers' ability to detect compressed images was examined for patterns; 95% CIs were used for statistical testing. RESULTS: With transilluminated film at clinical reading distance, readers were as likely to rate originals (48 [20%] of 240 readings) as degraded as they were to rate 20:1 replicates (106 [22%] of 480 readings) as degraded, but they frequently identified 50:1 replicates (283 [59%] of 480 readings) as degraded. At close inspection, 20:1 replicates (163 [34%] of 480 readings) were often identified as degraded, but 10:1 replicates (19 [8%] of 240 readings) were not identified as degraded more often than originals (17 [7%] of 240 readings). With CRT display, the results were nearly identical. CONCLUSION: At reading distance for primary interpretation, full-size digital chest radiographs that have been JPEG compressed to 10:1 or 20:1 and reconstructed are visually lossless at film or CRT display. Images compressed to 10:1 remain visually lossless at close inspection. PMID- 12893901 TI - Celiac artery compression by the median arcuate ligament: a pitfall of end expiratory MR imaging. AB - PURPOSE: To measure the prevalence and degree of celiac artery compression during breath-hold imaging at end inspiration and end expiration in patients referred to undergo magnetic resonance (MR) imaging of the abdomen for reasons unrelated to intestinal ischemia. MATERIALS AND METHODS: A series of 100 patients underwent routine MR imaging of the upper abdomen at 1.5 T; imaging included multiple dynamic contrast-enhanced fat-suppressed transverse three-dimensional spoiled gradient-echo acquisitions (3.6-4.5/1.5-1.9 [repetition time msec/echo time msec], 12 degrees flip angle). Arterial phase acquisitions were obtained during suspended respiration at end expiration (n = 50) or at end inspiration (n = 50), and venous phase acquisitions were obtained at the opposite respiratory phase. Two radiologists, blinded to patient identity and the phase of respiration, independently assessed the degree of narrowing on reconstructed oblique sagittal images. Radiologists reached consensus in 97 patients, who formed the cohort for this study. The percentage of stenosis of the celiac artery relative to its origin and the angle formed by the proximal celiac artery and the aorta were also measured in all patients. This angle and the arcsine transformation of the percentage of stenosis were compared for the two respiratory phases by using a paired Student t test. chi2 analysis was used to evaluate whether the degree of narrowing was independent of the breath-hold protocol that was used. RESULTS: In total, 55 (57%) of 97 patients had at least mild artery narrowing at end expiration, of whom 40 (73%) had less narrowing at end inspiration and 11 (20%) had no change. The average percentage of stenosis at end expiration (21% +/- 16) was significantly higher than that at end inspiration (11% +/- 11; P <.001). At end expiration, the average celiac artery angle was significantly lower in patients with mild to severe narrowing (41 degrees +/- 19) than in those without narrowing (50 degrees +/- 19; P <.03). CONCLUSION: Accentuation of celiac artery compression at end expiration can give rise to a potential pitfall of breath-hold abdominal imaging. When compression is suspected, imaging should be performed during inspiration. PMID- 12893903 TI - Case 61: ileocecal sarcoidosis. PMID- 12893904 TI - Hepatic MR imaging with ferumoxides: multicenter study of safety and effectiveness of direct injection protocol. AB - PURPOSE: To compare the safety and effectiveness of an undiluted direct injection of ferumoxides with those of a diluted slow infusion of ferumoxides during 30 minutes in patients with known liver lesions or in those suspected of having them. MATERIALS AND METHODS: Two hundred thirty-three patients at 16 institutions were randomized to receive either an undiluted direct injection of 0.56 mg of iron per kilogram of body weight of ferumoxides administered during 2 minutes (2 mL/min) or a diluted slow infusion administered during 30 minutes. Safety was assessed with monitoring for adverse events and laboratory tests. For sensitivity, specificity, and accuracy analysis, two independent blinded observers identified and classified lesions as benign or malignant with precontrast images and with pre- and postcontrast images combined. RESULTS: There was no statistically significant difference in adverse events in the group with direct injection compared with those in the group with infusion (21 [18%] of 114 patients vs 19 [17%] of 112 patients, respectively). No serious adverse events were observed. The most common adverse events in the group with direct injection versus the group with infusion were headache (five [4%] of 114 vs three [3%] of 112, respectively) and back pain (five [4%] of 114 vs three [3%] of 112, respectively). Overall, in 68 (62%) of 109 patients with direct injection and 71 (66%) of 108 patients with infusion, additional magnetic resonance (MR) imaging information was obtained after ferumoxides administration (P =.67). Sensitivity, specificity, and accuracy for the diagnosis of malignancy were significantly improved by adding images obtained after ferumoxides administration to the images obtained before contrast agent administration (P <.05 for all comparisons). CONCLUSION: Direct injection of ferumoxides has safety and effectiveness profiles similar to those of slow infusion of the agent. Further findings indicate that the addition of ferumoxides increases the sensitivity and specificity of hepatic MR evaluation when compared with unenhanced MR imaging. PMID- 12893905 TI - MR imaging of normal and matrix-depleted cartilage: correlation with biomechanical function and biochemical composition. AB - PURPOSE: To correlate articular cartilage function, as reflected in biomechanical properties and biochemical composition, with magnetic resonance (MR) imaging parameters of normal articular cartilage and cartilage partially depleted of matrix components. MATERIALS AND METHODS: Normal articular cartilage from 12 porcine patellae was evaluated biomechanically, biochemically, and with MR imaging (with and without gadolinium enhancement). The patellae were then enzymatically treated to deplete the matrix of either collagen or proteoglycan and then reevaluated biomechanically, biochemically, and with MR imaging. Correlations between cartilaginous tissue function and MR imaging parameters were made. Analysis of variance was performed to assess the effect of enzymatic treatment on measured parameters. Linear correlations among the MR imaging, biochemical, and biomechanical parameters were performed to determine the strengths of the relationships. P <.05 indicated statistically significant differences. RESULTS: Biochemical, biomechanical, and MR analyses enabled detection of changes caused by matrix depletion (P <.05). T2 was the most useful MR imaging parameter for distinguishing proteoglycan loss from collagen loss. T2 correlated significantly with both biomechanical modulus (indicative of cartilage stiffness; P <.001, R2 = 0.51) and biochemical proteoglycan content (P <.001, R2 = 0.44). Differentiation between proteoglycan loss and collagen loss in terms of T1 improved with gadolinium enhancement. With gadolinium enhancement, proteoglycan depletion was associated with a greater decrease in T1 than collagen depletion (P <.05). CONCLUSION: An association between biochemical and biomechanical functional status and MR imaging parameters of articular cartilage was demonstrated. Linear correlations existed between modulus and proteoglycan content in terms of T2. Additionally, proteoglycan loss and collagen loss had differing effects on gadolinium-enhanced T1 when it was expressed as the ratio of T1 after gadolinium enhancement/T1 before gadolinium enhancement. PMID- 12893906 TI - In vivo US monitoring of catheter-based vascular delivery of gene microspheres in pigs: feasibility. AB - In this study, the authors tested the feasibility of using ultrasonography (US) to monitor catheter-based vascular gene microsphere delivery. Polymeric biodegradable microspheres (mean diameter, 5 microm) were prepared by using a double-emulsion technique to encapsulate DNA-plasmid-encoding green fluorescent protein (GFP) genes. With use of gene-delivery catheters, GFP microspheres were locally delivered into the left femoral arterial walls of six pigs; the contralateral arteries were not infused with microspheres and thus served as negative control vessels. The delivery procedures were monitored with high frequency (8-15-MHz) transducer US. The effectiveness of monitoring with US was compared with the effectiveness of monitoring with immunohistochemical anti-GFP staining. A highly echogenic "star burst" sign around the entire vessel wall was seen at US and correlated with immunohistochemical findings that showed the destination of the gene microspheres. Study results demonstrate the potential of US for monitoring catheter-based vascular gene microsphere delivery in vivo. PMID- 12893907 TI - Image license from RSNA: a new threat to responsible authorship? PMID- 12893908 TI - Balloon dilation for achalasia of the cardia: a day case procedure. PMID- 12893909 TI - Ketamine-induced sedation. PMID- 12893910 TI - A challenge for the EU. PMID- 12893911 TI - Biomedical research. A low-stress scheme for overhauling NIH's structure. PMID- 12893912 TI - Global change. New climate science plan garners split opinions. PMID- 12893913 TI - Stem cells. Visiting German profs could face jail. PMID- 12893914 TI - Forensic science. Reformers aim to shake up British system. PMID- 12893915 TI - International health. U.S., Italy plan joint research effort. PMID- 12893916 TI - Physics. Quantum dots chemically wired for spintronics. PMID- 12893918 TI - A museum looted: mayhem in Mesopotamia. PMID- 12893917 TI - Public health. European Union plans for its own CDC. PMID- 12893919 TI - A museum looted: state board hopes for fresh start. PMID- 12893920 TI - Researchers weigh in on trading. PMID- 12893921 TI - Sites in hinterland present ripe targets for looters. PMID- 12893922 TI - Editor bias on climate change? PMID- 12893923 TI - Dating the Sterkfontein fossils. PMID- 12893924 TI - Politically correct or a bad decision? PMID- 12893925 TI - Early U.S. interest in hypersonic flight. PMID- 12893926 TI - Comment on "Obesity and the environment: where do we go from here?". PMID- 12893928 TI - Perceptions of science. Tap into science 24-7. PMID- 12893929 TI - Structural biology. Breaching the barrier. PMID- 12893930 TI - Physics. Getting entangled in free space. PMID- 12893931 TI - Physics. Getting a grip on turbulence. PMID- 12893932 TI - Development. There's something curious about paternal-age effects. PMID- 12893933 TI - Chemistry. Electrons as anions. PMID- 12893934 TI - Reconstruction of the proto-mitochondrial metabolism. PMID- 12893935 TI - Structure and mechanism of the lactose permease of Escherichia coli. AB - Membrane transport proteins that transduce free energy stored in electrochemical ion gradients into a concentration gradient are a major class of membrane proteins. We report the crystal structure at 3.5 angstroms of the Escherichia coli lactose permease, an intensively studied member of the major facilitator superfamily of transporters. The molecule is composed of N- and C-terminal domains, each with six transmembrane helices, symmetrically positioned within the permease. A large internal hydrophilic cavity open to the cytoplasmic side represents the inward-facing conformation of the transporter. The structure with a bound lactose homolog, beta-D-galactopyranosyl-1-thio-beta-D-galactopyranoside, reveals the sugar-binding site in the cavity, and residues that play major roles in substrate recognition and proton translocation are identified. We propose a possible mechanism for lactose/proton symport (co-transport) consistent with both the structure and a large body of experimental data. PMID- 12893936 TI - Structure and mechanism of the glycerol-3-phosphate transporter from Escherichia coli. AB - The major facilitator superfamily represents the largest group of secondary membrane transporters in the cell. Here we report the 3.3 angstrom resolution structure of a member of this superfamily, GlpT, which transports glycerol-3 phosphate into the cytoplasm and inorganic phosphate into the periplasm. The amino- and carboxyl-terminal halves of the protein exhibit a pseudo two-fold symmetry. Closed off to the periplasm, a centrally located substrate translocation pore contains two arginines at its closed end, which comprise the substrate-binding site. Upon substrate binding, the protein adopts a more compact conformation. We propose that GlpT operates by a single-binding site, alternating access mechanism through a rocker-switch type of movement. PMID- 12893937 TI - Surface-driven switching of liquid crystals using redox-active groups on electrodes. AB - Electrochemical control of the oxidation state of ferrocene-decorated electrodes leads to surface-driven changes in the orientations of thermotropic liquid crystals. When the electrodes possess nanometer-scale topography, voltages of 0.0 to 0.3 volts (versus a counter electrode in a two-electrode cell) can drive changes in the orientation of the liquid crystals in the plane and/or out of the plane of the electrodes. Electrodes not supporting ferrocene do not lead to surface-driven orientational transitions. The in-plane transitions are driven by the reorganization of the monolayer of ferrocene upon oxidation of ferrocene to ferrocenium. The out-of-plane transition reflects a dielectric coupling between the liquid crystal and the diffuse part of an electrical double layer that evolves upon oxidation of ferrocene to ferrocenium. These results suggest new ways to couple the orientations of liquid crystals to chemical and electrical stimuli in electro-optical devices and chemical sensors. PMID- 12893938 TI - High-density electron anions in a nanoporous single crystal: [Ca24Al28O64]4+(4e ). AB - We removed approximately 100% of clathrated oxygen ions from the crystallographic cages in a single crystal of 12CaO.7Al2O3, leading to the formation of high density (approximately 2 x 10(21) cm-3) electrons highly localized in the cages. The resulting electron forms a structure that we interpret as an F+ center and migrates throughout the crystal by hopping to a neighboring cage with conductivity approximately 100 siemens per centimeter, demonstrating that the encaged electron behaves as an anion. The electron anions couple antiferromagnetically with each other, forming a diamagnetic pair or singlet bipolaron. The resulting [Ca24Al28O64]4+(4e-) may be regarded as a thermally and chemically stable single crystalline "electride." PMID- 12893939 TI - Large trench-parallel gravity variations predict seismogenic behavior in subduction zones. AB - We demonstrate that great earthquakes occur predominantly in regions with a strongly negative trench-parallel gravity anomaly (TPGA), whereas regions with strongly positive TPGA are relatively aseismic. These observations suggest that, over time scales up to at least 1 million years, spatial variations of seismogenic behavior within a given subduction zone are stationary and linked to the geological structure of the fore-arc. The correlations we observe are consistent with a model in which spatial variations in frictional properties on the plate interface control trench-parellel variations in fore-arc topography, gravity, and seismogenic behavior. PMID- 12893940 TI - Extended Boltzmann kinetic equation for turbulent flows. AB - Complex fluid physics can be modeled using an extended kinetic (Boltzmann) equation in a more efficient way than using the continuum Navier-Stokes equations. Here, we explain this method for modeling fluid turbulence and show its effectiveness with the use of a computationally efficient implementation in terms of a discrete or "lattice" Boltzmann equation. PMID- 12893941 TI - Stratospheric memory and skill of extended-range weather forecasts. AB - We use an empirical statistical model to demonstrate significant skill in making extended-range forecasts of the monthly-mean Arctic Oscillation (AO). Forecast skill derives from persistent circulation anomalies in the lowermost stratosphere and is greatest during boreal winter. A comparison to the Southern Hemisphere provides evidence that both the time scale and predictability of the AO depend on the presence of persistent circulation anomalies just above the tropopause. These circulation anomalies most likely affect the troposphere through changes to waves in the upper troposphere, which induce surface pressure changes that correspond to the AO. PMID- 12893942 TI - Evidence for selective advantage of pathogenic FGFR2 mutations in the male germ line. AB - Observed mutation rates in humans appear higher in male than female gametes and often increase with paternal age. This bias, usually attributed to the accumulation of replication errors or inefficient repair processes, has been difficult to study directly. Here, we describe a sensitive method to quantify substitutions at nucleotide 755 of the fibroblast growth factor receptor 2 (FGFR2) gene in sperm. Although substitution levels increase with age, we show that even high levels originate from infrequent mutational events. We propose that these FGFR2 mutations, although harmful to embryonic development, are paradoxically enriched because they confer a selective advantage to the spermatogonial cells in which they arise. PMID- 12893943 TI - A conserved domain in axonal targeting of Kv1 (Shaker) voltage-gated potassium channels. AB - Axonal voltage-gated potassium (Kv1) channels regulate action-potential invasion and hence transmitter release. Although evolutionarily conserved, what mediates their axonal targeting is not known. We found that Kv1 axonal targeting required its T1 tetramerization domain. When fused to unpolarized CD4 or dendritic transferrin receptor, T1 promoted their axonal surface expression. Moreover, T1 mutations eliminating Kvbeta association compromised axonal targeting, but not surface expression, of CD4-T1 fusion proteins. Thus, proper association of Kvbeta with the Kv1 T1 domain is essential for axonal targeting. PMID- 12893944 TI - Reelin promotes peripheral synapse elimination and maturation. AB - Reelin is an extracellular protein that is crucial for layer formation in the embryonic brain. Here, we demonstrate that Reelin functions postnatally to regulate the development of the neuromuscular junction. Reelin is required for motor end-plate maturation and proper nerve-muscle connectivity, and it directly promotes synapse elimination. Unlike layer formation, neuromuscular junction development requires a function of Reelin that is not mediated by Disabled1 or very-low-density lipoprotein receptors and apolipoprotein E receptor 2 receptors but by a distinct mechanism involving its protease activity. PMID- 12893945 TI - Genome-wide insertional mutagenesis of Arabidopsis thaliana. AB - Over 225,000 independent Agrobacterium transferred DNA (T-DNA) insertion events in the genome of the reference plant Arabidopsis thaliana have been created that represent near saturation of the gene space. The precise locations were determined for more than 88,000 T-DNA insertions, which resulted in the identification of mutations in more than 21,700 of the approximately 29,454 predicted Arabidopsis genes. Genome-wide analysis of the distribution of integration events revealed the existence of a large integration site bias at both the chromosome and gene levels. Insertion mutations were identified in genes that are regulated in response to the plant hormone ethylene. PMID- 12893946 TI - Screening for nitric oxide-dependent protein-protein interactions. AB - Because nitric oxide (NO) may be a ubiquitous regulator of cellular signaling, we have modified the yeast two-hybrid system to explore the possibility of NO dependent protein-protein interactions. We screened for binding partners of procaspase-3, a protein implicated in apoptotic signaling pathways, and identified multiple NO-dependent interactions.Two such interactions, with acid sphingomyelinase and NO synthase, were shown to occur in mammalian cells dependent on endogenous NO. Nitrosylation may thus provide a broad-based mechanism for regulating interactions between proteins. If so, systematic proteomic analyses in which redox state and NO bioavailability are carefully controlled will reveal a large array of novel interactions. PMID- 12893947 TI - Combination drug therapy and mild hypothermia after transient focal cerebral ischemia in rats. AB - BACKGROUND AND PURPOSE: We have recently demonstrated that pretreatment with magnesium (calcium and glutamate antagonist) and tirilazad (antioxidant) in combination with intraischemic mild hypothermia (33 degrees C) (MTH) offers superior neuroprotective efficacy in a rat model of focal transient cerebral ischemia. In the present study, we investigated the time window of this treatment strategy with a posttreatment regimen to define its role for stroke patients. METHODS: We subjected 48 Sprague-Dawley rats to 90 minutes of middle cerebral artery occlusion by an intraluminal filament. Bilateral regional cerebral blood flow was continuously recorded by laser Doppler flowmetry. Combination therapy with MTH was started at 0, 1, 3, and 5 hours after induction of ischemia. Drugs were given in 1-hour intervals, and hypothermia was maintained for 2 hours. Neurological deficits were assessed daily. Infarct size was planimetrically determined on postoperative day 7. RESULTS: Combination therapy with MTH significantly reduced infarct volume compared with normothermic controls by -74%, -49%, and -45% when applied at 0, 1, and 3 hours after induction of ischemia. Furthermore, these treatment groups showed less neurological deficits on postischemic days 1 and 2 (P<0.05). Onset of treatment 5 hours after middle cerebral artery occlusion failed to significantly reduce infarct formation and neurological deficits. CONCLUSIONS: The therapeutic window of the new combination therapy is at least 3 hours after onset of ischemia, comparable to that of moderate hypothermia (30 degrees C), a grade of hypothermia associated with higher risks of severe side effects. PMID- 12893948 TI - External carotid artery territory ischemia impairs outcome in the endovascular filament model of middle cerebral artery occlusion in rats. AB - BACKGROUND AND PURPOSE: Middle cerebral artery occlusion (MCAO) by an intraluminal filament is a widely accepted animal model of focal cerebral ischemia. In this procedure, cutting of the external carotid artery (ECA) is a prerequisite for thread insertion. However, the implications of ECA transsection have not yet been described. METHODS: After 90 minutes of filament MCAO or sham surgery, rats were evaluated for up to 14 days in terms of body weight development, core temperature, and motor performance. Repeated in vivo MRI of the head and neck was performed for quantification of brain edema and infarct volume. The temporal muscles were histologically analyzed postmortem. RESULTS: In 47% of all rats, ischemic tissue damage to the ipsilateral ECA area, including temporal, lingual, and pharyngeal musculature, was detectable by MRI. Histology of temporal muscles confirmed acute ischemic myopathy. Animals with ECA territory ischemia (ECA-I) showed delayed body weight development and poorer recovery of motor function. There was no difference in the extent of brain edema or final cerebral lesion size between ECA-I-affected and unaffected rats. CONCLUSIONS: Filament MCAO was complicated by the consequences of ECA ischemia in approximately half of all rats. Impaired mastication and swallowing functions restricted ingestion and resulted in postsurgical body weight loss and worse motor performance. Impaired cerebral microperfusion resulting from dehydration and reduced spontaneous motor activity resulting from reduced food and water uptake might have contributed to poorer neurological recovery in ECA ischemic rats. Thus, adverse effects caused by extracerebral ischemia with potential impact on outcome have to be considered in this stroke model. PMID- 12893949 TI - QTc interval prolongation is a predictor of future strokes in patients with type 2 diabetes mellitus. AB - BACKGROUND AND PURPOSE: QTc interval prolongation is a predictor of cardiovascular morbidity and mortality in general populations and in patients with diabetes. The aim of this study was to investigate the predictors of stroke in patients with type 2 diabetes, with particular emphasis on the independent role of QT interval parameters. METHODS: We carried out a long-term follow-up study with 471 type 2 diabetics. Several clinical, laboratory, ECG, and echocardiographic variables were recorded at baseline. Predictive factors for stroke were evaluated by Kaplan-Meier estimation of survival curves and by univariate and multivariate Cox survival analyses. RESULTS: After a median follow up of 57 months (range, 2 to 84 months), 40 incident strokes were observed. QTc interval prolongation (>or=470 ms1/2) was an independent predictor of stroke, with adjusted hazard ratios ranging from 2.2 to 2.9 (95% confidence intervals, 1.1 to 6.0). Other independent factors associated with stroke were older age; the presence of cerebrovascular disease at baseline; increased 24-hour proteinuria, serum triglycerides, and left ventricular mass; and decreased high-density lipoprotein cholesterol. Excluding patients with previous cerebrovascular disease from the analysis did not change the results significantly. CONCLUSIONS: QTc interval prolongation is a predictor of future stroke in patients with type 2 diabetes. Intervention studies are needed to assess whether this factor could be modified. PMID- 12893950 TI - Levels of alpha1-antitrypsin in plasma and risk of spontaneous cervical artery dissections: a case-control study. AB - BACKGROUND AND PURPOSE: Abnormalities of dermal connective tissue have been detected in patients with spontaneous cervical artery dissections (sCAD), suggesting an underlying structural defect of the arterial wall. Alpha1 antitrypsin (A1-AT) is a circulating serine proteinase inhibitor of proteolytic enzymes that helps to maintain the integrity of elastic and collagen fibers. METHODS: To test the hypothesis that moderate deficiency of A1-AT may be a risk factor for sCAD, 22 cases with sCAD and 113 controls were included in the study. RESULTS: Patients with sCAD had significantly mean lower levels of A1-AT compared with controls (116.0+/-24.9 versus 141.1+/-31.7 mg/dL; P<0.01). Low levels of A1 AT (<90 mg/dL) were more frequently observed in patients with sCAD compared with controls (27.3% versus 2.7%; P<0.001). A positive correlation between age and plasma levels of A1-AT was found (r=0.22; P<0.01). A1-AT levels were not affected by sex or vascular risk factors, including smoking habit. On multivariate analysis, A1-AT <90 mg/dL was associated with sCAD independently of age, sex, or vascular risk factors (odds ratio, 17.7; 95% confidence interval, 2.9 to 105.6). CONCLUSIONS: Low plasma levels of A1-AT may be a risk factor for sCAD. PMID- 12893951 TI - Relationship between periodontal disease, tooth loss, and carotid artery plaque: the Oral Infections and Vascular Disease Epidemiology Study (INVEST). AB - BACKGROUND AND PURPOSE: Chronic infections, including periodontal infections, may predispose to cardiovascular disease. The present study investigates the relationship of periodontal disease and tooth loss with subclinical atherosclerosis. METHODS: We enrolled 711 subjects with a mean age of 66+/-9 years and no history of stroke or myocardial infarction in the Oral Infections and Vascular Disease Epidemiology Study. Subjects received a comprehensive periodontal examination, extensive in-person cardiovascular disease risk factor measurements, and a carotid scan using high-resolution B-mode ultrasound. Regression models were adjusted for conventional risk factors (age, sex, smoking, diabetes, systolic blood pressure, low- and high-density lipoprotein cholesterol, race-ethnicity, education, physical activity) and markers of cultural background, healthy lifestyle, and psychosocial health. RESULTS: Measures of both current and cumulative periodontitis became more severe as tooth loss increased. A significant association was observed between tooth loss levels and carotid artery plaque prevalence. Among those with 0 to 9 missing teeth, 46% had carotid artery plaque, whereas among those with >or=10 missing teeth, carotid artery plaque prevalence was approximately 60% (P<0.05). CONCLUSIONS: Our data suggest that tooth loss is a marker of past periodontal disease in this population and is related to subclinical atherosclerosis, thereby providing a potential pathway for a relationship with clinical events. PMID- 12893952 TI - Persistent poststroke hyperglycemia is independently associated with infarct expansion and worse clinical outcome. AB - BACKGROUND AND PURPOSE: Hyperglycemia at the time of ischemic stroke is associated with increased mortality and morbidity. Animal studies suggest that infarct expansion may be responsible. The influence of persisting hyperglycemia after stroke has not previously been examined. We measured the blood glucose profile after acute ischemic stroke and correlated it with infarct volume changes using T2- and diffusion-weighted MRI. METHODS: We recruited 25 subjects within 24 hours of ischemic stroke symptoms. Continuous glucose monitoring was performed with a glucose monitoring device (CGMS), and 4-hour capillary glucose levels (BGL) were measured for 72 hours after admission. MRI and clinical assessments were performed at acute (median, 15 hours), subacute (median, 5 days), and outcome (median, 85 days) time points. RESULTS: Mean CGMS glucose and mean BGL glucose correlated with infarct volume change between acute and subacute diffusion-weighted MRI (r>or=0.60, P<0.01), acute and outcome MRI (r=0.56, P=0.01), outcome National Institutes of Health Stroke Scale (NIHSS; r>or=0.53, P<0.02), and outcome modified Rankin Scale (mRS; r>or=0.53, P=0.02). Acute and final infarct volume change and outcome NIHSS and mRS were significantly higher in patients with mean CGMS or mean BGL glucose >or=7 mmol/L. Multiple regression analysis indicated that both mean CGMS and BGL glucose levels >or=7 mmol/L were independently associated with increased final infarct volume change. CONCLUSIONS: Persistent hyperglycemia on serial glucose monitoring is an independent determinant of infarct expansion and is associated with worse functional outcome. There is an urgent need to study normalization of blood glucose after stroke. PMID- 12893953 TI - Perfusion thresholds in acute stroke thrombolysis. AB - BACKGROUND AND PURPOSE: Perfusion-weighted MRI has been shown to be useful in the early identification of cerebral tissue at risk of infarction during acute ischemia. Identification of threshold perfusion measures that predict infarction may assist in the selection of patients for thrombolysis. METHODS: Mean transit time (MTT), regional cerebral blood flow (rCBF), and regional cerebral blood volume (rCBV) maps were generated in 35 acute stroke patients (17 treated with tissue plasminogen activator and 18 control patients) imaged within 6 hours from symptom onset. Day 90 outcome infarcts (T2-weighted MRI) were superimposed on acute MTT, rCBF, and rCBV maps. Perfusion-weighted MRI measures were then calculated for 2 regions: infarcted and salvaged tissue. RESULTS: MTT was prolonged by 22% in infarcted regions relative to salvaged tissue (P<0.001). rCBF was 10% lower in infarcted tissue than in salvaged regions (P<0.01). rCBV did not differ significantly between infarcted and salvaged regions. When reperfusion occurred, tissue with more severely prolonged MTT was salvaged from infarction relative to patients with persistent hypoperfusion (P<0.05). In contrast, rCBF in salvaged regions did not differ between patients with and without reperfusion. In reperfused patients, an inverse correlation (R=0.93, P<0.001) was found between time of initial MRI scan and MTT delay in salvaged tissue. CONCLUSIONS: Both increases in MTT and decreases in rCBF predict infarction. Differences in MTT also predict salvage in more severely hypoperfused tissue after reperfusion, suggesting that it is the most clinically useful quantitative perfusion measure. Perfusion thresholds for infarction need to be assessed in the context of symptom duration. PMID- 12893955 TI - Direction of foot force for pushes against a fixed pedal: role of effort level. AB - Control of the force exerted by the foot on the ground is critical to human locomotion. During running on a treadmill and pushing against a fixed pedal, humans increased foot force in a linear manner in sagittal plane force space. However, for pushes against a moving pedal, force output was linear for some participants but slightly curved for others. A primary difference between the static and dynamic pedaling studies was that the dynamic study required participants to push with varying peak effort levels, whereas a constant peak effort level was used for the fixed pedal pushes. The present study evaluated the possibility that force direction varied with level of effort. Seated humans pushed against a fixed pedal to a series of force magnitude targets. The force direction varied systematically with effort level consistent with the force path curvature observed for dynamic pedaling. PMID- 12893956 TI - Effects of two different dynamic environments on force adaptation: exposure to a new force but not the preceding force experience accounts for transition- and after-effects. AB - This study investigated force adaptation in humans during goal-directed flexion forearm motion. The ability of the motor system to adapt to changes in internal or external forces is essential for the successful control of voluntary movement. In a first experiment, we examined how under- or overdamping differentially affected the length of the adaptation and the arm kinematics between force transitions. We found that transitions diverging from a null-force produced larger transition effects than transitions converging to a null force condition, indicating that re-adaptation was less error-prone. Whether the subjects had previously experienced underdamping or the null-force had no significant impact on the spatial trajectory after switching to overdamping. That is, prior force experience had no differential effect on the spatial transition kinematics. However, the transitions underdamping-to-overdamping and underdamping-to-null force did produce differently strong transition effects. These results indicate that exposure to the new force rather than previous force-field experience is responsible for transition- and after-effects. In a second experiment, we investigated whether learning was law-like-that is, whether it generalized to unvisited workspace. Subjects were tested in new, unvisited workspaces in the null-force condition after sufficient training in either force condition. The occurrence of transferred after-effects indicated that adaptation to both positive and negative damping was mediated by rule-based rather than exclusive associative processes. PMID- 12893957 TI - The effect of changes in the body configuration on anticipatory postural adjustments. AB - A number of factors are likely to play a major role in the process of generation of anticipatory postural adjustments (APAs). Among them are the magnitude and direction of an expected perturbation, properties of a voluntary action associated with the perturbation, and features of the postural task such as a body's configuration prior to the action. The aim of this study was to analyze the effect of body configuration on APAs. Experiments were performed on 8 healthy subjects performing fast bilateral shoulder extension movements while standing. Body configuration was modified by instructions to the subjects to stand vertically or with a forward upper body bend varying from 15 to 60 degrees. The electrical activity of postural muscles and displacements of the center of pressure were recorded. Results indicated that APAs were modified with changes in the angular position of the upper body. Decreased anticipatory activation was seen in rectus abdominis and rectus femoris, while increased anticipatory inhibition was observed in erecteor spinae and biceps femoris across conditions with forward bend. As a result, the total anticipatory activity of muscles in a muscle pair in series with a forward bend showed only slight modulation as compared to vertical posture. These results suggest that the CNS uses reorganization of the anticipatory activity of postural muscles by compensating for the changes in APAs of individual muscles in a muscle pair in such a way that the overall anticipatory activity of the muscle pair stays unchanged. Such compensation in counteracting the expected mechanical effects of the perturbation is used to accommodate both changes in the length of postural muscles and diminished stability of the body due to forward bend. PMID- 12893958 TI - Effects of target height and width on 2D pointing movement duration and kinematics. AB - This study examined the impact of target geometry on the trajectories of rapid pointing movements. Participants performed a graphic point-to-point task using a pen on a digitizer tablet with targets and real time trajectories displayed on a computer screen. Circular- and elliptical-shaped targets were used in order to systematically vary the accuracy constraints along two dimensions. Consistent with Fitts Law, movement time increased as target difficulty increased. Analysis of movement kinematics revealed different patterns for targets constrained by height (H) and width (W). When W was the constraining factor, movements of greater precision were characterized by a lower peak velocity and a longer deceleration phase, with trajectories that were aimed relatively farther away from the center of the target and were more variable across trials. This indicates an emphasis on reactive, sensory-based control. When H was the constraining factor, however, movements of greater precision were characterized by a longer acceleration phase, a lower peak velocity, and a longer deceleration phase. The initial trajectory was aimed closer to the center of the target, and the trajectory path across trials was more constrained. This suggests a greater reliance on both predictive and reactive control. PMID- 12893959 TI - The utilization of visual feedback in the control of movement direction: evidence from a video aiming task. AB - The purpose of the present study was to establish the contribution of visual feedback in the correction of errors during movement execution (i.e., online) and the utilization of visual feedback from a completed movement in the programming of upcoming trials (i.e., offline). Participants performed 2 dimensional sweeping movements on a digitizing tablet through 1 of 3 targets, which were represented on a video monitor. The movements were performed with and without visual feedback under 4 criterion movement times (150, 250, 350, 450 msec). We analyzed the variability in directional error at 25%, 50%, 75%, and 100% of the distance between the home position and the target. There were significant differences in variability between visual conditions at each movement time. However, in the 150 msec condition, the form of the variability profiles did not differ between visual conditions, suggesting that the contribution of visual feedback was due to offline processes. In the 250-, 350-, and 450-msec conditions, there was evidence for both online and offline control, as the form of the variability profiles differed between the vision and no vision conditions. PMID- 12893960 TI - When motor execution is selectively impaired: control of manipulative finger forces in amyotrophic lateral sclerosis. AB - Amyotrophic lateral sclerosis is a degenerative motor neuron disorder with progressive and exclusive loss of motor neurons in the spinal cord, brainstem, and motor cortex. Five patients with amyotrophic lateral sclerosis, and 5 age matched, healthy control subjects performed vertical point-to-point arm movements with an instrumented hand-held object. In between the movements, the object was held stationary. Compared with healthy controls, all patients generated greater grip forces during the phase of stationary holding of the object and greater ratios between grip and load force maximums during the arm movements. We conclude that in amyotrophic lateral sclerosis, the ability to scale the grip force magnitude efficiently according to the actual loading requirements is impaired. When performing upward movements, controls increased grip force in parallel with load force right from the movement onset; during downward movements, controls anticipated an early decrease of load force by constant or decreasing grip forces. In contrast, 3 of 5 patients showed an early increase of grip force during both upward and downward movements, indicating that in amyotrophic lateral sclerosis, the differential regulation of the grip force output according to the direction-dependent load force profile may be impaired. In motor neuron disease, the inaccurate grip force scaling and the impaired temporal coupling between grip and load force profiles may either directly result from deficient motor execution or be secondary to accompanying symptoms, such as dyscoordination of hand and finger muscles due to spasticity. PMID- 12893961 TI - The fission yeast RPA51 is a functional homolog of the budding yeast A49 subunit of RNA polymerase I and required for maximizing transcription of ribosomal DNA. AB - Saccharomyces cerevisiae A49 and mouse PAF53 are subunits specific to RNA polymerase I (Pol I) in eukaryotes. It has been known that Pol I without A49 or PAF53 maintains non-specific transcription activities but a molecular role(s) of A49 (and PAF53) remains totally unknown. We studied the fission yeast gene encoding a protein of 415 amino acids exhibiting 30% and 19% identities to A49 and PAF53, respectively. We designate the corresponding protein RPA51 and gene encoding it rpa51+ since the gene encodes a Pol I subunit and an apparent molecular mass of the protein is 51 kDa. rpa51+ is required for cell growth at lower but not at higher temperatures and is able to complement S. cerevisiae rpa49Delta mutation, indicating that RPA51 is a functionally-conserved subunit of Pol I between the budding yeast and the fission yeast. Deletion analysis of rpa51+ shows that only two-thirds of the C-terminal region are required for the function. Transcripts analysis in vivo and in vitro shows that RPA51 plays a general role for maximizing transcription of rDNA whereas it is dispensable for non-specific transcription. We also found that RPA51 associates significantly with Pol I in the stationary phase, suggesting that Pol I inactivation in the stationary phase of yeast does not result from the RPA51 dissociation. PMID- 12893962 TI - Roles of sexual cell agglutination in yeast mass mating. AB - The agalpha1 mutant MAT alpha cells specifically lack the cell surface alpha-type sexual agglutination substance, which is also called alpha-agglutinin. Because the mutant cells (MATalpha agalpha1) can not form aggregates with MATa cells, MATalpha agalpha1 cells are unable to mate with MATa cells when they are co inoculated in a liquid medium, and the mating is attenuated on solid medium. The attenuated mating ability shown in the previous studies gave us a vague idea about a physiological function of the sexual agglutinability. In order to solve the question, mating behavior of MATalpha agalpha1 cells was investigated here under conditions where the contact between MATa and MAT alpha cells is assisted by physical methods. A synthetic mutation agalpha1::URA3 was constructed and used as well as agalpha1-1 for this study to ensure the genetic defect. When a mixture of MATa and MAT alpha cells was kept on filter membrane placed on relatively dry agar medium, even agalpha1::URA3 mutant cells mated as efficiently as the wild type (AGalpha1) cells did. On filter membrane placed on moist agar medium, agalpha1 mutants mated 10-fold less efficiently than wild type cells did. The mutant cells mated 10000-time less efficiently than the wild type cells in a pellet formed by brief low speed centrifugation. In contrast, the wild type MATalpha cells mated well under all conditions tested. Under the pellet condition, a mixture of MATa and MATalpha AG alpha1 cells formed an extended and cotton-like pellet while a mixture of MATa and MATalpha agalpha1 cells formed a compact and tight pellet. These results suggest that sexual cell agglutination contributes not only to cell contact between MATa and MAT alpha cells thereby stabilizing a-alpha cell pairs, but also to construction of a uniquely organized ultra structure favorable for zygote formation and subsequent growth of diploid cells. The mating specific extended pellet formation was observed also in 4 pairs of a and alpha strains in ascosporogenous yeast genera Hansenula and Pichia. PMID- 12893963 TI - Trimorphic DNA variation in the receptor-like protein kinase gene in the F18L15 130 region of the wild plant Arabidopsis thaliana. AB - DNA variation in the F18L15-130 region, which contains a receptor-like protein kinase gene, was analyzed for the wild plants Arabidopsis thaliana and Arabis gemmifera. In A. thaliana, at least three divergent sequence types were observed (trimorphism), instead of two distinct sequence types (dimorphism) detected in most of other nuclear gene regions studied for this plant species. Intragenic recombinations among the divergent sequence types generated four recombinant sequence types, although pattern of intragenic recombinations was complex. The estimated nucleotide variation in A. thaliana was the highest (pi = 0.0226 and theta = 0.0228) among genes investigated in this plant so far. The high level of nucleotide variation is due to divergence among the three distinct sequence types, each of which has a low level of nucleotide variation. Tests of Tajima, and Fu and Li did not detected deviation from neutrality, except for replacement sites, for which significantly negative Fu and Li's test value was detected. These results suggest that the receptor-like protein kinase gene in this region is functional. Possible causes for the trimorphic pattern of DNA polymorphism was discussed. PMID- 12893964 TI - Inheritance of Oryza sativa endornavirus in F1 and F2 hybrids between japonica and indica rice. AB - We have found a 14 kbp double-stranded RNA (dsRNA) in many cultivars of japonica rice (Oryza sativa L.) but not in any cultivars of indica rice. This dsRNA is an RNA replicon with plasmid-like properties and is proposed to be a novel dsRNA virus, Oryza sativa endornavirus (OSV). Reciprocal crosses between the OSV carrier japonica variety (Nipponbare) and the OSV-free indica variety (IR 26 or Kasalath) were performed to investigate whether OSV can be transmitted to F1 hybrids. When IR 26 and Nipponbare were used, efficient transmission of OSV from ova (93%) and pollen (89%) was observed. When Kasalath and Nipponbare were used, the OSV transmission efficiency to F1 progeny was 68% from ova and 20% from pollen. The transmission of OSV to F2 progeny plants was also complicated, showing non-Mendelian inheritance. These results suggest that the dsRNA replicon (OSV) is unstable in indica rice plants. PMID- 12893965 TI - Genetic variation in Oryza species detected by MITE-AFLP. AB - MITE-AFLP markers were successfully used to study the genetic variation and species relationship in Oryza species. Analysis of 53 accessions of Oryza species with seven MITE-AFLP primer combinations detected a total of 250 polymorphic fragments. High polymorphism was detected within and between Oryza species. Species relationships were analyzed by the pattern of presence or absence of homologous fragments, because nucleotide sequences of the detected MITE-AFLP fragments revealed that the same fragments in different species shared very high sequence homology. The genetic distances (GDs) between species were higher than those within species and the GDs in O. sativa complex were higher than those in O. officinalis complex. The phylogenetic tree recognized two major groups at 62% genetic similarity; group I consists of all AA genome species of the O. sativa complex, and group II consists of BB-, CC-, EE- and BBCC genome species of the O. officinalis complex. Therefore, this study demonstrated that the MITE-AFLP technique provide a tool for studying the genetic variation and species relationship in Oryza species. PMID- 12893966 TI - Accumulation of deleted mitochondrial DNA in aging Drosophila melanogaster. AB - Cumulative damage in mitochondria by reactive oxygen species is thought to result in a decrease in mitochondrial respiratory function and to contribute to the age related decline in the physiological function of organisms. The mitochondrial genome is also subjected to damage with age through deletions. The accumulation of deleted mitochondrial DNA (mtDNA) has been observed in various animals, but still remains unclear in insects. We examined the accumulation of deleted mtDNA in D. melanogaster at various ages from larvae to 65-day-old adults. When DNA extracted from whole bodies was examined by PCR and Southern hybridization, the age-related accumulation of deletions was not clear. However, when the accumulation of deleted mtDNA with age was examined separately in three parts of the body (head, thorax and abdomen), deleted mtDNA signals were detected more frequently in the thorax and the accumulation was age-dependent. Three of the deleted mtDNA were cloned, and the breakpoints of the deletions were identified. These results strongly suggest that deleted mtDNA accumulates in Drosophila with age in a tissue-specific manner. PMID- 12893967 TI - The Drosophila misfire gene has an essential role in sperm activation during fertilization. AB - The male sterile mutation, misfire (mfr), of Drosophila melanogaster is a novel paternal effect, fertilization defective mutant that effects sperm head decondensation. mfr sperm were motile, appeared normal morphologically and were transferred to the female during copulation. However, less than 0.1% of eggs laid by females mated to mfr males hatched. Although mfr sperm entered eggs at a high frequency (93%), 99% of the inseminated eggs did not initiate the first nuclear division. Unlike wild type fertilizing sperm, the position and shape of mfr sperm tails within the egg were not constant, but varied in a seemingly random manner. The heads of inseminating mutant sperm were always located near the surface of eggs just underlying the egg plasma membrane, and maintained their needle-like shape indicating the failure of nuclear decondensation. Further observations revealed that plasma membrane of inseminating sperm appeared intact, including the head region. These phenotypes were equivalent to those of sneaky (snky), another fertilization defective male sterile mutation. Our observations strongly suggest that mfr mutant males are sterile because their inseminating sperm fail to form a male pronucleus due to the inability of the sperm to properly respond to egg factors responsible for the breakdown of the plasma membrane. Although mfr and snky mutations were phenotypically identical, they mapped to cytologically distinct genetic loci and no genetic interactions were observed, suggesting that at least two distinct paternal gene products are involved in the early stages of pronuclear formation. PMID- 12893968 TI - Fukuyama-type congenital muscular dystrophy (FCMD) and alpha-dystroglycanopathy. AB - Fukuyama-type congenital muscular dystrophy (FCMD), Walker-Warburg syndrome (WWS), and muscle-eye-brain (MEB) disease are clinically similar autosomal recessive disorders characterized by congenital muscular dystrophy, lissencephaly, and eye anomalies. Through positional cloning, we identified the gene for FCMD and MEB, which encodes the fukutin protein and the protein O-linked mannose beta1, 2-N-acetylglucosaminy ltransferase (POMGnT1), respectively. Recent studies have revealed that posttranslational modification of alpha-dystroglycan is associated with these congenital muscular dystrophies with brain malformations. In this review Fukuyama-type congenital muscular dystrophy (FCMD), other CMDs with brain malformations, and their relation with alpha-dystroglycan are discussed. PMID- 12893969 TI - Brain slice culture for analysis of developmental brain disorders with special reference to congenital cytomegalovirus infection. AB - Cytomegalovirus (CMV) is the most significant infectious cause of congenital abnormalities of the central nervous system (CNS) with variation from the fatal cytomegalic inclusion disease to functional brain disorder. The phenotype and degree of the brain disorder depends on infection time during the developing stage, virulence, route of infection and the viral susceptibility of the cells. The pathogenesis of the CMV infection to the CNS seems to be strongly related to neural migration, neural death, cellular compositions and the immune system of the brain. To understand the complex mechanism of this disorder, we used organotypic brain slice cultures. In the brain slice culture system, migration of CMV-infected neuronal cells was observed, which reflects infectious dynamics in vivo. Neural progenitor cells or glial immature cells in the subventricular zone and marginal area are most susceptible to murine cytomegalovirus (MCMV) infection in this system. The susceptibility declined as the number of immature glial cells decreased with age. The immature glial cells proliferated in brain slice cultures during prolonged incubation, and the susceptibility to MCMV infection also increased in association with the proliferation of these cells. The brain slice from an immunocompromised mouse (Beige-SCID mouse) unexpectedly showed lower susceptibility than that of an immunocompetent mouse during any prolonged incubation. These results suggest that the number of immature glial cells might determine the susceptibility of CMV infection to the brain, independent of the immune system. We reviewed recent findings of CMV infection to the brain from the perspective of brain slice cultures and the possibility that this system could be a useful method to investigate mechanisms of congenital anomaly of the brain. PMID- 12893970 TI - The dopaminergic system in attention deficit/hyperactivity disorder. AB - Numerous studies have shown the importance of the mesocorticolimbic dopamine system in the pathophysiology of attention deficit/hyperactivity disorder. However, there has been inconsistency in the findings of those studies. Varied and sometimes contradictory interpretation has been made on the basis of similar results. It is, therefore, still unclear whether the dopaminergic system is hypo- or hyperfunctioning in attention deficit/hyperactivity disorder. The majority of the functional brain imaging studies in both clinical and experimental settings support hypofunction of the basal ganglia which receive abundant dopaminergic afferent. The experimental studies, using dopamine-depleted animals, also support the hypodopaminergic hypothesis, whereas recent studies with the dopamine transporter knockout/knockdown mouse suggest hyperdopaminergic function as the underlying abnormality. In this review we attempt to clarify the issues raised by previous neuroimaging and functional neuroimaging studies. Research involving animal models with genetic traits, genetic manipulation or with brain lesions is analysed, concentrating on the significance of the dopaminergic system in the abnormal behavior of attention deficit/hyperactivity disorder. In addition, the functional state of the dopaminergic system is considered through the speculated mechanism of psychostimulant therapy of the disorder. No final conclusions have been reached regarding the pathological, biochemical and physiological mechanisms responsible for various symptoms. Inconsistency in the findings and their interpretations may indicate the heterogeneity of the pathogenesis of this syndrome. PMID- 12893971 TI - Zebrafish as a novel experimental model for developmental toxicology. AB - It is widely believed that embryos and infants during development are highly sensitive to chemicals that cause serious damage to growth. However, knowledge on the mechanisms of developmental toxicity is scarce. One reason for this is limited convenient model system other than organ cultures using rodents to study the various aspects of developmental toxicology. Cultured cells are not always adequate for this purpose, since events in morphogenesis are processed through interactions with other tissues. We focused on zebrafish embryo (Danio rerio), one of the most important organisms in developmental biology. Saturation mutagenesis, applied to drosophila and nematode to define the functions of genes, has been carried out in zebrafish but almost no other vertebrate, and several thousand lines are available due to the rapid growth and transparent body of this embryo. Enhanced databases for the genome and ESTs are available at websites with abundant genetic and biological background. By targeted gene knock-down with morpholino-modified antisense oligonucleotieds (morpholinos), the translation of a specific protein can be transiently blocked for several days. Many reporter systems in vivo have been established mainly as GFP-transgenic fish for environmental chemicals. Although several excellent studies have been performed with zebrafish embryos on the effects of chemicals, the developmental toxicology of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) has been most extensively studied to date. We have found that TCDD induces apoptosis in dorsal midbrain with a concomitant decrease in local blood flow, using developing zebrafish. TCDD seems to produce oxidative stress through CYP1A induction in vascular endothelium, resulting in local circulation failure and apoptosis in the dorsal midbrain. In addition to applications in toxicology, an experimental system with zebrafish embryos could help to clarify the mechanism of congenital anomaly, which arises from genetic mutation. PMID- 12893972 TI - A genetic mouse model carrying the nonfunctional xeroderma pigmentosum group G gene. AB - A genetic mouse model with a disrupted XPG allele was generated by insertion of neo cassette sequences into exon 3 of the XPG gene by using embryonic stem (ES) cell techniques. The xpg-deficient mice showed distinct developmental characteristics. Their body was marked smaller than that in wild-type littermates since the postnatal day 6, and this postnatal growth failure became more severe with developmental proceeding. Their life span was very short, all of the mutants died by postnatal day 23 after showing great weakness and emaciation. In addition, the mutant homozygous mice also showed some progressive neurological signs, like the lower level of activity and a progressive ataxia. Further examination indicated there was developmental retardation of the brain in the mutant mice. Their brain weight, and thickness of cerebral cortex and cerebellar cortex were significant different from the controls. These characteristics, like small size brain, brain developmental retardation and progressive neurological dysfunctions in the homozygotes were similar to the typical clinical phenotype of the XPG patients and Cockayne syndrome, we believe that the xpgdeficient mice will be an animal model for studying the function of the XP-G protein in nucleotide-excision repair and mechanisms related to the clinic symptoms of XP-G and Cockayne syndrome in humans. PMID- 12893973 TI - Predictors of women's attendance at cardiac rehabilitation programs. AB - This descriptive study was conducted to identify the factors that influence womens attendance at cardiac rehabilitation programs and womens adherence to risk factor modification following a cardiac event. Women (N=196) admitted to hospital for a cardiac event were followed-up at 12 weeks postdischarge. Despite eligibility, only 64% (n=112) had been referred to cardiac rehabilitation programs. By 12 weeks postdischarge only 32% of the total sample (n=57) attended programs and 12% of the total sample (n=21) had dropped out before completion. The odds of a woman attending cardiac rehabilitation were decreased by myocardial infarction diagnosis, lack of employment, <55 years or >70 years, and experiencing a personal stressful event during follow-up. Women were likely to adhere to smoking, medication, and stress modification guidelines but unlikely to adhere to modification guidelines for diet and exercise. PMID- 12893974 TI - Perceptions of activity and vocational status in women with cardiac illness. AB - The purpose of this descriptive study was to determine the perceptions of activity and vocational status in women with cardiac illness. A convenience sample of 20 women with the diagnosis of coronary heart disease or hypertrophic cardiomyopathy participated. Subjects underwent an audiotaped interview, using a semistructured interview guide, which was then transcribed into the Ethnograph software program (Qualis Research Associates, Salt Lake City, UT). Transcripts were analyzed for common themes via content analysis. There were four themes identified for activity: variable activity patterns, fatigue, guilt, and depression. There were four themes identified for vocational status: desire to work, social aspects of work, unpaid work, and struggle to maintain the status quo. In conclusion, the themes of fatigue, guilt over unmet activity expectations, and the common occurrence of unpaid work validated prior research findings. The themes of variable patterns of activity management, feelings of depression about lowered activity levels, the desire to return to a prior vocation, the importance of the social nature of work, and the struggle to balance both home and work roles were new findings. PMID- 12893975 TI - Coronary heart disease risk in people 65 years of age and older. AB - Evidence from epidemiologic studies indicates that the same factors that are associated with increased risk of coronary heart disease (CHD) in middle-aged people are relevant in older adults (i.e., those aged >or=65). The relative risk associated with some risk factors decreases with advancing age but this is offset by greater incidence of CHD among older adults. A growing body of evidence from clinical trials indicates that risk factor modification in older adults reduces CHD risk as effectively as it does in middle-aged adults. Multivariable risk assessment can be used to effectively target intervention to those at significant risk for an initial CHD event and to avoid over-treatment. It is important to appreciate that the average remaining life expectancy after achieving 80 years is about 8 years. PMID- 12893976 TI - Noncompliance in heart transplantation: a role for the advanced practice nurse. AB - Many organ transplant recipients experience organ rejection because they are noncompliant with the requirements of the health care regimen. The advanced practice nurse is in an ideal position to assess predictors of noncompliance as well as to implement interventions to enhance patient compliance. The purpose of this paper is to present a case study of a young female heart transplant recipient whose death due to organ rejection was related to noncompliance. The role of the advanced practice nurse in reducing noncompliance is identified and relevant nursing interventions are discussed. PMID- 12893977 TI - Cardiovascular nursing in Zambia. PMID- 12893978 TI - Cardiovascular effects of migraine therapy. PMID- 12893979 TI - Why is the QT interval measurement so critical when evaluating an ECG rhythm? PMID- 12893980 TI - Physiological effects of simvastatin and diet on hypercholesterolemic men. PMID- 12893981 TI - The single nucleotide polymorphism story. PMID- 12893982 TI - Functional significance of a hereditary adenine insertion variant in the 5'-UTR of the endothelin-1 gene. AB - Endothelin-1 (ET-1) is known as a potent vasoconstrictor peptide and stimulator of cell proliferation. The human preproendothelin-1 mRNA contains a frequent adenine insertion polymorphism (allele frequency = 0.28) within the 5' untranslated region (5'-UTR), 138 bp downstream of the transcription start site, which was assumed to be related to hypertension. This 5'-UTR variant could putatively influence the mRNA secondary structure and stability, efficacy of translation initiation, or binding of sequence-specific mRNA-binding proteins. By cloning the entire ET-1 gene 5'-UTR in a pGL3 vector and transfection of two cell lines, we studied the effects on luciferase expression in vitro. Luciferase activity was significantly increased in the insertion variant (I) compared to the wild-type (D) variant for both COS1 (2.97 +/- 0.12 versus 2.17 +/- 0.10; P = 0.002) and HepG2 cells (5.42 +/- 0.90 versus 3.68 +/- 0.37; P = 0.002). Investigations performed ex vivo using human umbilical vein endothelial cells were performed to examine the influence of genotypes on the formation of mRNA and protein. Preproendothelin-1-mRNA was quantified in relation to GAPDH by a realtime polymerase chain reaction. Homozygous I-carriers showed significant elevated mRNA levels compared to I/D and I/I-carriers (I/I 9.03 +/- 1.86, I/D 2.07 +/- 1.15, D/D 2.33 +/- 0.99; P = 0.001). ET-1 protein expression, determined by enzyme-linked immunosorbent assay, was increased among I-carriers (I/I 814 +/- 144, I/D 528 +/- 103, D/D 556 +/- 75 pg/ml; P = 0.001). The observed effects may be due to an enhanced mRNA stability because the half-life of mRNA consisting of the I-variant was prolonged (35.4 +/- 7.9 versus 19.9 +/- 4.5 min). We were able to show that the +138 I/D polymorphism is of functional importance for ET-1 expression, and this may have consequences for vessel tonus regulation. PMID- 12893983 TI - Prediction of successful weight reduction under sibutramine therapy through genotyping of the G-protein beta3 subunit gene (GNB3) C825T polymorphism. AB - BACKGROUND: Sibutramine, a centrally acting noradrenaline and serotonin re-uptake inhibitor, enhances satiety and is frequently used to support weight loss. However, a significant variability exists among individuals concerning the response to sibutramine. METHODS: We genotyped 111 participants of a randomized placebo-controlled clinical trial for the GNB3 C825T polymorphism and analysed associations of genotypes with treatment outcome. Patients undergoing a structured weight loss programme were treated with either placebo or 15 mg sibutramine daily for 54 weeks. RESULTS: In the placebo group, the non pharmacological programme alone resulted in a significantly greater weight loss in individuals with the GNB3 TT/TC genotypes as compared to individuals with the CC genotype (-7.1 +/- 1.2 vs. -2.7 +/- 1.5 kg, P = 0.031). Administration of 15 mg sibutramine was more effective in individuals with the CC genotype than in the subjects with the TT/TC genotypes (weight loss: 7.2 +/- 2.2 vs. 4.1 +/- 2.1 kg, P = 0.0013, sibutramine vs. placebo). In the CC genotype carriers, the odds ratio (OR) for a weight loss greater than 5% (sibutramine vs. placebo) was 6.6 (95% CI 1.8-25.6; P = 0.004) and for a weight loss greater than 10% was 9.6 (95% CI 1.7 53.8; P = 0.010). CONCLUSION: Genotyping for the GNB3 C825T polymorphism is highly predictive for the identification of obese individuals who will benefit from sibutramine treatment. PMID- 12893984 TI - Genetic findings and functional studies of human CYP3A5 single nucleotide polymorphisms in different ethnic groups. AB - OBJECTIVES: Genetic polymorphisms of cytochromes P450 (CYPs) are a principal reason for inter-individual variations in the metabolism of therapeutic drugs and environmental chemicals in humans. The present study identifies 34 single nucleotide polymorphisms (SNPs) of CYP3A5 including 27 previously unidentified SNPs by direct sequencing of the exons, intron-exon junctions and 5'-upstream region of CYP3A5 from 92 racially diverse individuals (24 Caucasians, 24 Africans, 24 Asians, and 20 individuals of unknown racial origin). RESULTS: Four new CYP3A5 SNPs produced coding changes: R28C, L82R, A337T, and F446S. CYP3A5 R28C occurred in African populations (allelic frequency of 4%). CYP3A5 A337T occurred in Asians (2% allelic frequency), CYP3A5 L82R (occurred in the racially unidentified group) and CYP3A5 F446S (identified in Caucasians with a 2% allelic frequency) were on an allele containing the splice change g.6986A>G known as CYP3A5*3. The newly identified allelic proteins were constructed by site-directed mutagenesis, expressed in Escherichia coli and purified. CYP3A5 L82R was expressed only as denatured CYP420, suggesting it may be unstable. CYP3A5*1 exhibited the highest maximal clearance for testosterone followed by CYP3A5 A337T > CYP3A5 R28C >> CYP3A5 F446S. CYP3A5*1 exhibited a higher V(max) for nifedipine oxidation than CYP3A5 A337T > CYP3A5 R28C >> CYP3A5 F446S. CYP3A5 A337T and CYP3A5 R28C exhibited a 42-64% lower V(max) for nifedipine oxidation than CYP3A5*1. CYP3A5 F446S exhibited a > 95% decrease in the intrinsic clearance for both 6beta-hydroxytestosterone and nifedipine oxidation. CONCLUSION: This study identifies four new potentially defective coding alleles. CYP3A5 F446S is predicted to be more catalytically defective than the splice change alone. PMID- 12893985 TI - Influence of CYP2C9 genetic polymorphisms on pharmacokinetics of celecoxib and its metabolites. AB - In-vitro data indicate major effects of the genetically polymorphic cytochrome P450 enzyme 2C9 (CYP2C9) on the pharmacokinetics of celecoxib, a nonsteroidal anti-inflammatory drug acting as selective cyclooxygenase-2 inhibitor. Human studies report decreased clearance in heterozygous carriers of the CYP2C9 variant Ile359Leu (*3), but results appeared controversial and only data on single subjects carrying the homozygous CYP2C9*3/*3 genotype have been published. We measured single-dose kinetics of celecoxib and its main metabolites hydroxy- and carboxy-celecoxib in 21 healthy volunteers who were selected as hetero- (n = 4) and homozygous (n = 3) carriers of CYP2C9 variants Arg144Cys (*2) and Ile359Leu (*3). Blood concentrations of celecoxib and its metabolites hydroxy-celecoxib and carboxy-celecoxib were quantified by high-performance liquid chromatography. A more than two-fold reduced oral clearance in homozygous carriers of CYP2C9*3 was seen for celecoxib compared to carriers of the wild-type genotype CYP2C9*1/*1 and heterozygous carriers of one *3 allele were in-between (P = 0.003 for trend), whereas CYP2C9*2 had no significant influence on celecoxib pharmacokinetics. Decreased concentrations of carboxy- and hydroxy-celecoxib in heterozygous and homozygous carriers of CYP2C9*3 were detected which supported the influence of CYP2C9 polymorphisms on celecoxib pharmacokinetic variability. Approximately 0.5% of Caucasians carrying the genotype CYP2C9*3/*3 will have greatly increased internal exposure to celecoxib. It remains to be shown whether this is associated with greater efficacy or with an increased incidence and severity of adverse events. PMID- 12893986 TI - Sequence diversity and haplotype structure in the human ABCB1 (MDR1, multidrug resistance transporter) gene. AB - OBJECTIVES: There is increasing evidence that polymorphism of the ABCB1 (MDR1) gene contributes to interindividual variability in bioavailability and tissue distribution of P-glycoprotein substrates. The aim of the present study was to (1) identify and describe novel variants in the ABCB1 gene, (2) understand the extent of variation in ABCB1 at the population level, (3) analyze how variation in ABCB1 is structured in haplotypes, and (4) functionally characterize the effect of the most common amino acid change in P-glycoprotein. METHODS AND RESULTS: Forty-eight variant sites, including 30 novel variants and 13 coding for amino acid changes, were identified in a collection of 247 ethnically diverse DNA samples. These variants comprised 64 statistically inferred haplotypes, 33 of which accounted for 92% of chromosomes analyzed. The two most common haplotypes, ABCB1*1 and ABCB1*13, differed at six sites (three intronic, two synonymous, and one non-synonymous) and were present in 36% of all chromosomes. Significant population substructure was detected at both the nucleotide and haplotype level. Linkage disequilibrium was significant across the entire ABCB1 gene, especially between the variant sites found in ABCB1*13, and recombination was inferred. The Ala893Ser change found in the common ABCB1*13 haplotype did not affect P glycoprotein function. CONCLUSION: This study represents a comprehensive analysis of ABCB1 nucleotide diversity and haplotype structure in different populations and illustrates the importance of haplotype considerations in characterizing the functional consequences of ABCB1 polymorphisms. PMID- 12893987 TI - Deleterious mutations in the flavin-containing monooxygenase 3 (FMO3) gene causing trimethylaminuria. AB - The primary genetic form of trimethylaminuria (TMAU) is caused by inherited defects in the flavin-containing monooxygenase 3 (FMO3) gene. Defective FMO3 has a decreased ability to catalyze the N-oxygenation of the dietary-derived malodourous amine, trimethylamine. We report two novel deleterious mutations identified in two unrelated individuals affected by the disorder. Sequence analysis of the FMO3 coding exons amplified from genomic DNA revealed that the mutation from individual 1 was heterozygous for a G>A missense mutation in exon 2 of the FMO3 gene. The mutation changed a GAG encoding Glu at codon 32 to AAG encoding Lys. Wild-type and mutant E32K FMO3 were expressed in Escherichia coli as maltose binding-fusion proteins and assayed for their ability to catalyze oxygenation of various FMO3 substrates. The results showed that the E32K mutation abrogated the catalytic activity of the enzyme. Individual 2 was identified as heterozygous for the P153L mutation. In addition, individual 2 was also heterozygous for a novel single nucleotide deletion of A191 in exon 3 at codon 64. The deletion resulted in a frame shift and caused premature termination of the FMO3 gene immediately after codon 65. Family pedigree analysis revealed that the P153L and the deletion mutation were carried on different alleles for this individual. Therefore, both alleles of the FMO3 gene for individual 2 were affected by mutations abolishing the catalytic activity of the enzyme, explaining the severe TMAU condition. The two deleterious mutations reported herein were rare mutations with estimated allelic frequencies of less than 1%. PMID- 12893988 TI - Genotyping for polymorphic drug metabolizing enzymes from paraffin-embedded and immunohistochemically stained tumor samples. AB - OBJECTIVES: Paraffin-embedded tumor samples are valuable in the study of cancer for routine staging, tumor marker analysis, and in retrospective studies to test new prognostic and predictive biomarkers. Their utility in retrospective pharmacogenetic analysis of clinical trials has yet to be evaluated. We set out to establish genotyping methods for relevant genes from archival tumor samples and determine if fixation, processing or somatic changes in the tumor might affect our ability to identify germ-line polymorphisms. METHODS AND RESULTS: To establish the assays, paraffin blocks were made using pellets prepared from eight tumor cell lines. DNA was isolated from viable cells and from sections from these blocks, and genotyped for polymorphisms in CYP2C8, CYP2C9, CYP2C19, CYP2D6, CYP3A5 and MDR1 using conventional PCR-RFLP assays. This demonstrated that fixation and processing did not alter the genotypes obtained (100% concordance). Next, sections were obtained from paraffin-embedded archival breast samples from 10 patients for whom gDNA isolated from peripheral blood was available for comparison. Concordance was complete with the same genotype being obtained for 100% of the samples tested. Attempts to extend these methods for the study of hematoxylin/eosin or immunohistochemically stained sections were not successful since the staining inhibited the PCR reactions. Only 25 of 50 samples were successfully amplified and of those only 14 produced accurate genotypes. CONCLUSIONS: Accurate genetic testing for polymorphisms in several genes of pharmacogenetic importance can be obtained from archival paraffin-embedded tumor samples. Thus, pharmacogenetic analysis can be applied to existing cancer therapy trials to test associations between these polymorphisms and treatment response. PMID- 12893989 TI - Mutations in mitochondrial aldehyde dehydrogenase (ALDH2) change cofactor affinity and segregate with voluntary alcohol consumption in rats. AB - Genetic factors influence alcohol consumption and alcoholism. A number of groups have bred alcohol drinker and non drinker rat strains, but genetic determinants remain unknown. The University of Chile rat lines UChA (low drinkers) and UChB (high drinkers) display differences in the relative K(m) for NAD+ of mitochondrial aldehyde dehydrogenase (ALDH2) but no V(max) differences. The relative K(m) differences may be due to mitochondrial changes or to genetic differences coding for ALDH2. We investigated whether there are differences in the coding regions of ALDH2 cDNA in these lines and whether the Aldh2 genotype predicts the phenotype of alcohol consumption and the K(m) of ALDH2 for NAD+. Liver cDNA was prepared, and the Aldh2 transcript was amplified, cloned and sequenced. Genotyping was conducted by DNA amplification and restriction enzyme digestion. When compared to Aldh21 of Sprague-Dawley, 94% of the UChA (low drinker) rats (n = 61), presented a mutation that changes Gln67 to Arg in the mature enzyme (allele referred to as Aldh22). In UChB (high drinker) rats (n = 69), 58% presented the Aldh21 allele, while 42% presented the Gln67Arg change plus a second mutation that changed Glu479 to Lys (allele Aldh23). The Aldh22 allele was absent in high drinker rats. Rats of different Aldh2 genotypes displayed marked phenotypic differences in both ethanol consumption (g/kg/day; means +/- SE): (Aldh21/Aldh21) = 5.7 +/- 0.2, (Aldh22/Aldh22) = 0.9 +/- 0.2 and (Aldh23/Aldh23) = 4.6 +/- 0.2; and K(m)s for NAD+ of 43 +/- 3 microm, 132 +/- 13 microm and 41 +/- 2 microm, respectively (Aldh22 versus Aldh21 or Aldh23; P < 0.0001 for both phenotypes). Overall, the data show that alleles of Aldh2 strongly segregate with the phenotype of ethanol consumption and the relative K(m) for NAD+ of ALDH2. Bases mutated suggest that non drinker Aldh22 is ancestral with regard to the coding changes in either Aldh21 or Aldh23, variants which would allow ethanol consumption and may provide an evolutionary advantage by promoting calorie intake from fermented products along with carbohydrates. PMID- 12893990 TI - UGT pharmacogenomics: implications for cancer risk and cancer therapeutics. AB - UDP-glucuronosyltransferases (UGTs) belong to a superfamily of microsomal enzymes responsible for glucuronidation of numerous endogenous and exogenous compounds including bilirubin, hormones, various drugs as well as environmental carcinogens. Glucuronidation predominantly serves as a pathway for elimination of the different glucuronidated compounds. Seventeen human UGT transcripts have been identified thus far, and the UGT proteins are differentially expressed in a wide range of human tissues. Genetic variants have been identified in coding and non coding sequences of several UGT genes, and similar observations should be anticipated for all UGTs. As glucuronidation plays a critical part in the inactivation or elimination of countless substrates, genetic variants in this enzyme family that lead to altered expression or activity of UGTs are likely to have some physiologic and pharmacological consequences. This article focuses on the potential impact of various UGTs or their variants on cancer risk and cancer therapeutics. PMID- 12893991 TI - Management of pharmacologically induced neuromuscular blockade using peripheral nerve stimulation. AB - Currently, many critically ill patients are treated with neuromuscular blocking agents to facilitate endotracheal intubation, mechanical ventilation, and manage confused, combative, head injured, and status epileptus patients. To prevent persistent pharmacologically induced paralysis, the aide of a peripheral nerve stimulator and clinical indicators may help determine the minimal effective drug dose to facilitate safe neuromuscular blockade. The purpose of this article is to provide nursing clinicians the essential knowledge to manage patients with neuromuscular blockade effectively using peripheral nerve stimulation monitoring. PMID- 12893992 TI - What's new in pulmonary artery hypertension. AB - Pulmonary hypertension is a debilitating disease and has a poor prognosis. This article provides an overview of new treatments for this disease. PMID- 12893993 TI - How to do an effective poster presentation in the workplace. PMID- 12893994 TI - Death, dying, and the ICU nurse. AB - Death is a natural progression from life. Most nurses will be exposed to the physical and emotional effects of this experience as they care for a dying patient. The nurse is taught how to provide support for the patient and family as they proceed through the stages of grief. Often, however the nurse may not realize his or her own need to grieve. PMID- 12893995 TI - A personal reflection: the adversarial relationship between physicians and nurse patients. PMID- 12893996 TI - Effects of relaxation and imagery on the sleep of critically ill adults. AB - The purpose of this study was to determine the effects of relaxation and imagery on the sleep of critically ill adults. The study was an experimental clinical trial with random assignment to two groups. Analysis used repeated measures ANOVA. Thirty-six adults (17 males and 19 females) with a variety of physical diagnoses in three critical-care units in two large metropolitan hospitals were studied. Outcome measures were scores on a visual analog sleep scale, measured on three mornings. The intervention was a combination of relaxation and imagery, delivered on two evenings. All subjects' sleep improved over time. There were significant interaction effects between the intervention, gender, and time, with males' scores improving rapidly, and females' scores first dropping, then improving rapidly. A combination of relaxation and imagery is effective in improving the sleep of the critically ill adult, with men responding immediately to relaxation and imagery with improved sleep, and women taking more time to respond to the intervention. PMID- 12893997 TI - A closing word: an RN as a second degree. PMID- 12893998 TI - Cardiac rehabilitation in community settings. PMID- 12893999 TI - Leisure time physical activity of patients in maintenance cardiac rehabilitation. AB - PURPOSE Increasing caloric expenditure through physical activity is associated with reduced mortality. On the basis of observational studies, previous authors have suggested that at least 1000 kcal per week and possibly 1500 kcal per week of physical activity is necessary for health benefits. The authors have previously reported that patients in maintenance cardiac rehabilitation accumulate approximately 230 kcal per exercise session, suggesting that additional activity outside of cardiac rehabilitation is needed to achieve the goal of 1500 kcal per week. The authors estimated the amount of energy expenditure performed each week by patients in cardiac rehabilitation during both program participation and leisure time. METHODS For this study, 104 patients enrolled in a supervised maintenance cardiac rehabilitation program at both tertiary care and community settings for at least 6 months completed a self administered physical activity questionnaire. Energy expenditure in cardiac rehabilitation and leisure time activity was measured in kilocalories. Total caloric expenditure was determined by adding up the number of kilocalories expended by the patients each week climbing stairs, walking, participating in cardiac rehabilitation, and engaging in sports. RESULTS Patients in cardiac rehabilitation expended weekly, on the average, 1504 +/- 830 kcal in physical activity, 830 +/- 428 kcal in cardiac rehabilitation, and 675 +/- 659 kcal in leisure time activity. There was a significant difference in caloric expenditure between men and women, between those with a body mass index (BMI) less than 30 and those with a BMI of 30 or more, and between those younger than 70 years and those 70 years or older. There was no difference between races. Whereas 43% of the patients accumulated 1500 kcal, 57% did not. CONCLUSIONS The findings showed that 72% of the patients in cardiac rehabilitation accumulated at least 1000 kcal of energy expenditure per week and met public health guidelines. Also, 43% of the patients in cardiac rehabilitation accumulated more than 1500 kcal of energy expenditure per week, a level identified as necessary to reduce all-cause mortality. Women of either race, patients with a BMI of 30 or more, and patients age 70 years or older are the groups least likely to achieve 1500 kcal of energy expenditure per week. The authors recommend incorporating weekly kilocalories of energy expenditure in the exercise prescription of patients to ensure achievement of maximum health benefits. PMID- 12894000 TI - In or out-it all adds up. PMID- 12894001 TI - The Enhancing Recovery in Coronary Heart Disease Trial (ENRICHD): strategies and techniques for enhancing retention of patients with acute myocardial infarction and depression or social isolation. AB - The report aims to review the literature and describe the methods used for retention of patients in a clinical study. The Enhanced Recovery in Coronary Heart Disease (ENRICHD) trial was a multicenter, randomized clinical trial designed to evaluate the effects of a psychosocial intervention on cardiovascular morbidity and mortality. A total of 2481 patients met the criteria for depression, low social support, or both after a myocardial infarction and needed to be followed. Follow-up evaluation consisted of telephone interviews 3, 9, 12, 24, 36, and 48 months after enrollment and clinic visits scheduled at 6, 18, 30, 42, and 54 months. Creative strategies used to achieve optimum retention of this complex patient population over a long follow-up period are presented. Strategies to enhance adherence throughout the course of the trial required adequate tracking of patients to ensure minimum dropout, follow-up evaluation optimized through multiple methods of contact to guarantee completeness of data collection; and development of procedures to address the needs of patients at risk for dropout. Patients in the group that completed the study participated for a mean of 28.3 months, and those lost to follow-up evaluation participated for a mean of 19 months. Retention was not substantially different by gender or minority status. The results of this project can assist investigators in planning studies that require patient follow-up evaluation, and can provide clinicians with specific strategies for maximizing retention-to-treatment recommendations. As a result of the retention strategies described in this report, 93.02% of the patients completed their study participation or died. This is a very high retention rate given the complexity of the study sample, protocol, and required duration of follow-up evaluation. PMID- 12894002 TI - Factors associated with the failure of patients to complete cardiac rehabilitation for medical and nonmedical reasons. AB - PURPOSE: Cardiac rehabilitation (CR) provides effective secondary prevention services, but many patients fail to complete the recommended program. The purposes of this study were to describe completion rates in a hospital-based outpatient CR program, and to identify factors associated with patients failing to complete CR because of nonmedical and medical reasons. METHODS: Data used for the analyses were from a hospital-based CR program involving 526 discharged patients between January 1996 and February 2002. Patient discharge status was classified into three categories: complete, noncomplete-medical reasons, and noncomplete-nonmedical reasons. Logistic regression modeling identified factors associated with the groups failing to complete CR. RESULTS: The rate of CR completion was 58% (304/526). Among the 222 patients who did not complete CR, 139 (63%) had nonmedical reasons. As compared with the patients who completed CR, the adjusted odds ratio (AOR) for those who did not complete CR because of nonmedical reasons were more likely to be employed (AOR 2.2), to be obese (AOR 2.5), to be smokers (AOR 2.1), and to have shorter 6-minute walk distances (AOR 1.7). They were less likely to be women (AOR 0.6) or have diabetes (AOR 0.5). Patients not completing CR for medical reasons were more likely to be categorized as being at high clinical risk (AOR 4.2) and having shorter 6-minute walk distances (AOR 1.9). CONCLUSION: Except for low functional capacity, baseline factors associated with patients failing to complete CR differed on the basis of medical or nonmedical reasons. The development of interventions that address the special needs of patients with low functional capacity may be especially important in attempts to retain this high-risk group in CR therapy. PMID- 12894003 TI - The Wisconsin outcomes experience: baseline outcomes of the WISCVPR Web-based Outcomes Project. AB - PURPOSE: The measurement of outpatient cardiac rehabilitation (OCR) outcomes is now considered common practice. Unfortunately, because of the costs involved in purchasing outcomes tracking software, many programs are not able to participate in pooled outcomes analysis and benchmarking. The purpose of this report is to describe the current OCR outcomes data in the state of Wisconsin using a web based model for outcomes data collection, data pooling, and benchmarking. METHODS: Participating OCR programs collected individual patient outcomes according to protocols developed by the Wisconsin Society for Cardiovascular and Pulmonary Rehabilitation's Outcomes Committee. Programs analyzed their data quarterly and submitted the values to a database via an Internet web site. Aggregate outcomes data from 34 programs were collected over a 27-month period. RESULTS: Aggregate data representing 5235 OCR patients are presented. The mean age for the men was 64 +/- 5 years (N = 3612; 69% of the sample). The mean age for the women was 67 +/- 6 years (N = 1623; 31% of the sample). The completion rate for the group was 89% (95%CI, 86-91). The average number of OCR sessions per patient was 22 +/- 6 over 9 +/- 3 weeks. The clinical and behavioral outcomes for this sample ranged from a median value of 88% for the percentage of patients meeting the blood pressure goal (95%CI, 87-91) to less than 5% for the percentage of patients receiving formal psychosocial counseling (95%CI, 0). The major cardiovascular event rate was 0.17 per 10,000 patient-hours. CONCLUSION: This report describes the baseline OCR outcomes obtained via a state affiliate sponsored outcomes management program. These data show that this model of OCR outcomes measurement, collection, and interprogram analysis produces data that are comparable with those of published, controlled studies. Determining baseline characteristics of OCR outcomes data will be important in the development of benchmarks and best practice guidelines. PMID- 12894004 TI - Quality of life and self-efficacy: gender and diagnoses considerations for management during cardiac rehabilitation. AB - PURPOSE: Outcome measurement research has extended beyond traditional clinical and physiologic parameters to include psychosocial aspects. Accordingly, the purpose of this study was to investigate quality-of-life (QOL) and self-efficacy disparities for gender and diagnoses during participation in cardiac rehabilitation. METHODS: For this study, 472 patients (114 women and 358 men) were stratified by gender and then again by diagnosis to include surgical revascularization, myocardial infarction, and percutaneous coronary intervention. Measures obtained at baseline and at the end of the study assessed quality of life (QOL-o = total score), including emotional (QOL-e) and limitation (QOL-l) domains; self-efficacy (SE-o = total score), including ambulatory (SE-a) and muscular (SE-m) domains; and caloric expenditure. RESULTS: Both self-efficacy and QOL were greater at the end of the study across genders (P <.05). The men had greater self-efficacy values for all domains (P <.05). There was a significant gender-time interaction for QOL-e (P <.05) among the women, and for QOL-o, QOL-l, and all self-efficacy domains (P <.05) among the surgical revascularization patients. Percutaneous coronary intervention patients had higher self-efficacy scores throughout. Caloric expenditure was a consistent positive predictor of self-efficacy and QOL-e (P <.05). CONCLUSIONS: Quality of life and self-efficacy improve during cardiac rehabilitation across gender and diagnoses. Female and revascularized patients present with low QOL and self-efficacy scores initially, but improvements in scores similar to or greater than the men can be expected. Because the self-efficacy scores of percutaneous coronary intervention patients are higher and their physical limitations are less prohibitive, these patients can be progressed more aggressively. Improvements in self-efficacy scores parallel caloric expenditure increases. PMID- 12894005 TI - Noninvasive ventilation during exercise training improves exercise tolerance in patients with chronic obstructive pulmonary disease. AB - PURPOSE: In patients with chronic obstructive pulmonary disease, pulmonary rehabilitation has been demonstrated to increase exercise capacity and reduce dyspnea. In the most disabled patients, the intensity of exercise during the training sessions is limited by ventilatory pump capacity. This study therefore evaluated the beneficial effect of noninvasive ventilation (NIV) support during the rehabilitation sessions on exercise tolerance. METHODS: This study included 14 patients with stabilized chronic obstructive pulmonary disease, ages 63 +/- 7 years, with a forced expiratory volume in 1 second (FEV(1)) 31.5% +/- 9.2% of predicted value. All 14 patients participated in an outpatient pulmonary rehabilitation program. Seven of the patients trained with NIV during the exercise sessions (NIV group), whereas the remaining seven patients breathed spontaneously (control group). Exercise tolerance was evaluated during an incremental exercise test and during constant work rate exercise at 75% of peak oxygen consumption (VO(2)) before and after the training program. RESULTS: The application of noninvasive ventilation increased exercise tolerance, reduced dyspnea, and prevented exercise-induced oxygen desaturation both before and after training. The pressure support was well tolerated by all the patients during the course of the training program. In the NIV group, training induced a greater improvement in peak VO(2) (18% vs 2%; P <.05) and a reduced ventilatory requirement for maximal exercise, as compared with the control group. The constant work rate exercise duration increased similarly in both groups (116% vs 81%, nonsignificant difference), and posttraining blood lactate was decreased at isotime (P <.05 in both groups), but not at the end of the exercise. CONCLUSION: In this pilot study, exercise training with noninvasive ventilation support was well tolerated and yielded further improvement in the increased exercise tolerance brought about by pulmonary rehabilitation in patients with chronic obstructive pulmonary disease. This improved exercise tolerance is partly explained by a better ventilatory adaptation during exercise. PMID- 12894006 TI - Hospital volume, length of stay, and readmission rates in high-risk surgery. AB - OBJECTIVE: Aimed at reducing surgical deaths, several recent initiatives have attempted to establish volume-based referral strategies in high-risk surgery. Although payers are leading the most visible of these efforts, it is unknown whether volume standards will also reduce resource use. METHODS: We studied postoperative length of stay and 30-day readmission rate after 14 cardiovascular and cancer procedures using the 1994-1999 national Medicare database (total n = 2.5 million). We used regression techniques to examine the relationship between length of stay, 30-day readmission, and hospital volume, adjusting for age, gender, race, comorbidity score, admission acuity, and mean social security income. RESULTS: Mean postoperative length of stay ranged from 3.4 days (carotid endarterectomy) to 19.6 days (esophagectomy). There was no consistent relationship between volume and mean length of stay; it significantly increased across volume strata for 7 of the 14 procedures and significantly decreased across volume strata for the other 7. Mean length of stay at very-low-volume and very-high-volume hospitals differed by more than 1 day for 6 procedures. Of these, the mean length of stay was shorter in high-volume hospitals for 3 procedures (pancreatic resection, esophagectomy, cystectomy), but longer for other procedures (aortic and mitral valve replacement, gastrectomy). The 30-day readmission rate also varied widely by procedure, ranging from 9.9% (nephrectomy) to 22.2% (mitral valve replacement). However, volume was not related to 30-day readmission rate with any procedure. CONCLUSION: Although hospital volume may be an important predictor of operative mortality, it is not associated with resource use as reflected by length of stay or readmission rates. PMID- 12894007 TI - Is it defensible to use volume standards for purchasing care? PMID- 12894008 TI - The aging population and its impact on the surgery workforce. AB - OBJECTIVE: To predict the impact of the aging population on the demand for surgical procedures. SUMMARY BACKGROUND DATA: The population is expanding and aging. According to the US Census Bureau, the domestic population will increase 7.9% by 2010, and 17.0% by 2020. The fastest growing segment of this population consists of individuals over the age of 65; their numbers are expected to increase 13.3% by 2010 and 53.2% by 2020. METHODS: Data on the age-specific rates of surgical procedures were obtained from the 1996 National Hospital Discharge Survey and the National Survey of Ambulatory Surgery. These procedure rates were combined with corresponding relative value units from the Centers for Medicare and Medicaid Services. The result quantifies the amount of surgical work used by an average individual within specific age groups (<15 years old, 15-44 years old, 45-64 years old, 65+ years old). This estimate of work per capita was combined with population forecasts to predict future use of surgical services. RESULTS: Based on the assumption that age-specific per capita use of surgical services will remain constant, we predict significant increases (14-47%) in the amount of work in all surgical fields. These increases vary widely by specialty. CONCLUSIONS: The aging of the US population will result in significant growth in the demand for surgical services. Surgeons need to develop strategies to manage an increased workload without sacrificing quality of care. PMID- 12894009 TI - "I prefer old age to the alternative.": Maurice Chevalier, 1962. PMID- 12894010 TI - Current concepts in the mediastinal lymph node staging of nonsmall cell lung cancer. AB - OBJECTIVE: To review the current concepts in the mediastinal staging of nonsmall cell lung cancer (NSCLC), evaluating traditional and modern staging modalities. SUMMARY BACKGROUND DATA: Staging of NSCLC includes the assessment of mediastinal lymph nodes. Traditionally, computed tomography (CT) and mediastinoscopy are used. Modern staging modalities include magnetic resonance imaging (MRI), positron emission tomography (PET), and endoscopic ultrasound with fine-needle aspiration (EUS-FNA) METHODS: Literature was searched with PubMed and SUMSearch for original, peer-reviewed, full-length articles. Studies were evaluated on inclusion criteria, sample size, and operating characteristics. Endpoints were accuracy, safety, and applicability of the staging methods. RESULTS: CT had moderate sensitivities and specificities. With few exceptions magnetic resonance imaging (MRI) offered no advantages when compared with CT, against higher costs. PET was significantly more accurate than CT. Mediastinoscopy and its variants were widely used as gold standard, although meta-analyses were absent. Percutaneous transthoracic needle biopsy (PTNB) and transbronchial needle biopsy (TBNA) were moderately sensitive and specific. EUS-FNA had high sensitivity and specificity, is a safe and fast procedure, and is cost-effective. EUS-FNA evaluates largely a nonoverlapping mediastinal area compared with mediastinoscopy. CONCLUSIONS: PET has the highest accuracy in the mediastinal staging of NSCLC, but is not generally used yet. EUS-FNA has the potential to perform mediastinal tissue sampling more accurate than TBNA, PTNB, and mediastinoscopy, with fewer complications and costs. Although promising, EUS-FNA is still experimental. Mediastinoscopy is still considered as gold standard for mediastinal staging of NSCLC. PMID- 12894011 TI - Efficacy of an anterior as compared with a posterior laparoscopic partial fundoplication: results of a randomized, controlled clinical trial. AB - OBJECTIVE: The aim of the study was to compare the efficacy and mechanical consequences of 2 partial fundoplications performed laparoscopically under the framework of a randomized, controlled clinical trial. SUMMARY BACKGROUND DATA: Although laparoscopic total fundoplication procedures have proven their effectiveness in the control of gastroesophageal reflux, problems remain with the functional consequences after a supra-competent gastric cardia high-pressure zone. Partial fundoplications have been found to be associated with fewer mechanical side effects. PATIENTS AND METHODS: During a 2-year period, 95 patients with gastroesophageal reflux disease were enrolled into a randomized, controlled single-institution clinical trial comparing a partial posterior (Toupet, n = 48) fundoplication and an anterior partial wrap (Watson, n = 47). All patients were assessed postoperatively at predefined time points, and the 12 month follow-up data are presented in terms of clinical results and 24-hour pH monitoring variables. RESULTS: Both patient groups were strictly comparable at the time of randomization. All operations were completed laparoscopically, and no serious complications were encountered. During the first postoperative year, a difference regarding the control of reflux symptoms was observed in favor of the posterior fundoplication. Esophageal acid exposure (% time pH <4) was substantially reduced by both operations but to a significantly lower level after a Toupet compared with the Watson partial fundoplication (1.0 +/- 0.3 vs. 5.6 +/- 1.1 mean +/- SEM; p < 0.001). Postfundoplication symptoms were infrequently recorded with no difference between the groups. CONCLUSIONS: When performing a laparoscopic partial fundoplication, the posterior modification (Toupet) offers advantages in terms of better reflux control compared with an anterior type (Watson). PMID- 12894012 TI - Significant host- and tumor-related factors for predicting prognosis in patients with esophageal carcinoma. AB - OBJECTIVE: To identify clinically useful parameters obtainable before treatment of predicting clinical outcomes in patients with esophageal carcinoma. SUMMARY BACKGROUND DATA: Various factors regarding the biologic state of tumors or the nutritional state of patients have been individually reported to correlate with prognosis. Reliable estimates of life expectancy before treatment are important, and consideration needs to be given not only to tumor-related but also to host related factors in patients with esophageal carcinoma. METHODS: The following clinicopathological factors were retrospectively analyzed in 356 consecutive patients with surgical treatment: sex; age; serum C-reactive protein (CRP); proportion of lymphocytes; body weight changes; serum albumin; clinical TNM staging; tumor location; serum squamous cell-related antigen; serum carcinoembryonic antigen; and histology. Factors related to prognosis were evaluated by using univariate and multivariate analyses. RESULTS: According to univariate analysis, significant differences in survival were found for sex, serum CRP, proportion of lymphocytes, body weight change, serum albumin, serum squamous cell-related antigen, and clinical TNM staging. Multivariate analysis demonstrated that CRP levels (P = 0.0285), body weight change (P = 0.0165), and clinical TNM staging (P = 0.0008) displayed independent correlations to prognosis. When serum CRP elevation, body weight loss, and clinical TNM staging III and IV were scored as a combined index, the total score (prognostic index for esophageal cancer, PIEC) demonstrated a good stratification value for prognosis. Moreover, PIEC was superior to the conventional clinical TNM staging by the likelihood ratio test. CONCLUSIONS: PIEC based on serum CRP, body weight change, and clinical TNM staging before treatment offers a very simple and informative method for predicting the prognosis of patients with esophageal carcinoma. PMID- 12894013 TI - Quality of life in rectal cancer patients: a four-year prospective study. AB - OBJECTIVE: To assess long-term quality of life in a population-based sample of rectal cancer patients. SUMMARY BACKGROUND DATA: Quality of life in rectal cancer patients who suffer reduced bowel and sexual function is very important. Few studies, however, have long term follow-up data or sufficient sample sizes for reliable comparisons between operation groups. PATIENTS AND METHODS: A 4-year prospective study of rectal cancer patients' quality of life was assessed by using the European Organization for Research and Treatment of Cancer QLQ-30 and CR38 questionnaires. RESULTS: A total of 329 patients returned questionnaires. Overall, anterior resection patients had better quality of life scores than abdominoperineal extirpation patients. High-anterior resection patients had significantly better scores than both low-anterior resection and abdominoperineal extirpation patients. Low-anterior resection patients, however, overall had a better quality of life than abdominoperineal extirpation patients, especially after 4 years. Abdominoperineal extirpation patients' quality of life scores did not improve over time. Stoma patients had significantly worse quality of life scores than nonstoma patients. Quality of life improved greatly for patients whose stoma was reversed. CONCLUSIONS: Anterior resection and nonstoma patients, despite suffering micturition and defecation problems, had better quality of life scores than abdominoperineal extirpation and stoma patients. Comparisons between abdominoperineal extirpation and anterior resection patients should consider the effect of temporary stomas. Improvements in quality of life scores over time may be explained by reversal of temporary stomas or physiologic adaptation. PMID- 12894014 TI - Similar outcome after colonic pouch and side-to-end anastomosis in low anterior resection for rectal cancer: a prospective randomized trial. AB - OBJECTIVES: To compare a colonic J-pouch or a side-to-end anastomosis after low anterior resection for rectal cancer with regard to functional and surgical outcome. SUMMARY BACKGROUND DATA: A complication after restorative rectal surgery with a straight anastomosis is low-anterior resection syndrome with a postoperatively deteriorated anorectal function. The colonic J-reservoir is sometimes used with the purpose of reducing these symptoms. An alternative method is to use a simple side-to-end anastomosis. METHODS: One-hundred patients with rectal cancer undergoing total mesorectal excision and colo-anal anastomosis were randomized to receive either a colonic pouch or a side-to-end anastomosis using the descending colon. Surgical results and complications were recorded. Patients were followed with a functional evaluation at 6 and 12 months postoperatively. RESULTS: Fifty patients were randomized to each group. Patient characteristics in both groups were very similar regarding age, gender, tumor level, and Dukes' stages. A large proportion of the patients received short-term preoperative radiotherapy (78%). There was no significant difference in surgical outcome between the 2 techniques with respect to anastomotic height (4 cm), perioperative blood loss (500 ml), hospital stay (11 days), postoperative complications, reoperations or pelvic sepsis rates. Comparing functional results in the 2 study groups, only the ability to evacuate the bowel in <15 minutes at 6 months reached a significant difference in favor of the pouch procedure. CONCLUSIONS: The data from this study show that either a colonic J-pouch or a side-to-end anastomosis performed on the descending colon in low-anterior resection with total mesorectal excision are methods that can be used with similar expected functional and surgical results. PMID- 12894015 TI - Prospective, age-related analysis of surgical results, functional outcome, and quality of life after ileal pouch-anal anastomosis. AB - OBJECTIVE To evaluate how age affects functional outcome and quality of life after ileal pouch anal anastomosis (IPAA). SUMMARY BACKGROUND DATA Because of the limited number of older patients undergoing IPAA, it has been difficult to assess functional outcome and quality of life stratified by age. METHODS IPAA was performed in 1895 patients. Patients were stratified by age into <45 (n = 1410), 46-55 (n = 289), 56-65 (n = 154), and more than 65 years (n = 42). Outcome was assessed prospectively. Results are presented at 1, 3, 5, and 10 years after surgery.RESULTS Patients were followed for 4.6 +/- 3.7 years (maximum, 17 years). Pouch failure occurred in 4.1% (pouch excision or permanent diversion). Incontinence and night time seepage were more common in older patients. There were minor differences in the quality of life, health, energy and happiness between age groups, with a slight benefit for those under 45 years. Fourteen percent or fewer patients experienced social, sexual or work restrictions. Overall, 96% of patients were happy to have undergone their surgery, and 98% recommended it to others. Although the respective figures were 89% and 96% in the over-65 age group, the difference was not significant. CONCLUSIONS These data provide a unique assessment of outcome after IPAA at multiple time points. Although functional outcome after IPAA is not as good in older patients, appropriate case selection confers acceptable function and quality of life to patients of all ages. PMID- 12894016 TI - Long-term failure after restorative proctocolectomy for ulcerative colitis. AB - OBJECTIVE: To establish the incidence and causes of late failure in patients undergoing restorative proctocolectomy for a preoperative diagnosis of ulcerative colitis was the objective of this investigation. SUMMARY BACKGROUND DATA: Restorative proctocolectomy is the elective surgical procedure of choice for ulcerative colitis. Most patients have a satisfactory outcome but failures occur. The reasons and rates of early failure are well documented, but there is little information on long-term failure. METHODS: A series of 634 patients (298 females, 336 males) underwent restorative proctocolectomy for inflammatory bowel disease between 1976-1997, with a mean follow-up of 85 +/- 58 months. Failure was defined as removal of the pouch or the need for an indefinite ileostomy. It was divided into early and late, occurring within 1 year or more than 1 year postoperatively. RESULTS There were 3 (0.5%) postoperative deaths, leaving 631 patients for analysis. Of these, 23 subsequently died (disseminated large bowel cancer, 12; unrelated causes, 9; related causes, 2). There were 61 (9.7%) failures (15 early [25%], 46 late [75%]) due to pelvic sepsis (32 [52%]: 7 early, 25 late), poor function (18 [30%]: 2 early, 16 late), pouchitis (7 [11%]: 2 early, 5 late) and miscellaneous (4, all early). A final diagnosis of Crohn's disease, type of reservoir (J,S), female gender, postoperative pelvic sepsis and a one-stage procedure were significantly associated with failure. Failure rate rose with time of follow-up from 9% at 5 years to 13% at 10 years. CONCLUSIONS: Pelvic sepsis and poor function were the main reasons for later failure. Failure rates should be reported based on the duration of follow-up. PMID- 12894017 TI - Massive splenomegaly is associated with significant morbidity after laparoscopic splenectomy. AB - OBJECTIVE: To evaluate the impact of spleen weight on operative and clinical outcome in a series of 108 consecutive laparoscopic splenectomies. BACKGROUND: Laparoscopic splenectomy as an alternative to open splenectomy for splenomegaly is regarded as controversial. METHODS: Patients underwent laparoscopic splenectomy for a range of hematological disorders between November 1992 and February 2000. Multiple linear and logistic regression analysis were used to assess the effect of massive splenomegaly (>1000 g) on perioperative mortality and morbidity, after adjusting for the joint effects of patient age, weight, pre- and postoperative full blood counts, operating time, estimated blood loss, conversion rate, reoperation rate, and duration of hospital stay. RESULTS: Massive splenomegaly was recorded in 27 of 108 (25%) cases. In this group, splenic weight ranged from 1000 to 4750 g (median, 2500 g). Patients with splenic weight >1000 g had a significantly longer median operating time (170 vs. 102 minutes, P < 0.01), conversion rate (5/27 vs. 4/81, P < 0.05), postoperative morbidity (15/27 vs. 4/81, P < 0.01), and median postoperative stay (5 vs. 3 days, P < 0.01). Multivariate analysis found splenic weight to be the most powerful predictor of morbidity (P < 0.01). Patients with splenomegaly (>1000 g) were 14 times likely to have post operative complications. One patient died 3 days after surgery, following a pulmonary embolus (spleen weight 500 g, mortality 1/108, 0.9%). CONCLUSIONS: Laparoscopic splenectomy is feasible in patients with giant spleens. However, it is associated with greater morbidity, and the advantages of minimal access surgery in this subgroup of patients are not so clear. PMID- 12894018 TI - Real-time quantification of AFP mRNA to assess hematogenous dissemination after transarterial chemoembolization of hepatocellular carcinoma. AB - OBJECTIVE: To determine whether the number of hepatocytes containing AFP mRNA shed into the bloodstream during transarterial chemoembolization (TAE) affects the incidence and pattern of recurrence of hepatocellular carcinoma (HCC). PATIENTS AND METHODS: We developed a Taqman procedure to quantify AFP mRNA prospectively in 52 consecutive patients before and after TAE. Results are expressed in hepatocytes /mL. RESULTS: Thirteen of the patients (24.5%) were positive for AFP mRNA (42 +/- 19 hepatocytes/mL) before TAE and 13 (24.5%) (80 +/ 32 hepatocytes/mL) after TAE; the difference was not significant. The presence of AFP mRNA in the bloodstream before TAE was associated with larger nodules (85.2 +/- 73.8 mm versus 34.8 +/- 26.1 mm; P = 0.006). Six of the patients were excluded from the analysis because they underwent curative surgery or were lost to follow-up. The circulating levels of AFP mRNA released in the 46 remaining patients after TAE did not affect metastasis-free survival. A significant number of extrahepatic metastases were found in patients exhibiting at least 1 AFP mRNA positive blood sample either before or after TAE. However, the TAE procedure did not increase the risk of extrahepatic recurrences. CONCLUSION: Cells containing AFP mRNA are inconsistently released into the circulation during TAE. The amount of these cells released does not affect the recurrence of HCC. PMID- 12894019 TI - Hypertonic saline enhances host response to bacterial challenge by augmenting receptor-independent neutrophil intracellular superoxide formation. AB - OBJECTIVE: This study sought to determine whether hypertonic saline (HTS) infusion modulates the host response to bacterial challenge. METHODS: Sepsis was induced in 30 Balb-C mice by intraperitoneal injection of Escherichia coli (5 x 107 organisms per animal). In 10 mice, resuscitation was performed at 0 and 24 hours with a 4 mL/kg bolus of HTS (7.5% NaCl), 10 animals received 4 mL/kg of normal saline (0.9% NaCl), and the remaining animals received 30 mL/kg of normal saline. Samples of blood, spleen, and lung were cultured at 8 and 36 hours. Polymorphonucleocytes were incubated in isotonic or hypertonic medium before culture with E. coli. Phagocytosis was assessed by flow cytometry, whereas intracellular bacterial killing was measured after inhibition of phagocytosis with cytochalasin B. Intracellular formation of free radicals was assessed by the molecular probe CM-H(2)DCFDA. Mitogen-activated protein (MAP) kinase p38 and ERK 1 phosphorylation, and nuclear factor kappa B (NFkappaB) activation were determined. Data are represented as means (SEM), and an analysis of variance test was performed to gauge statistical significance. RESULTS: Significantly reduced bacterial culture was observed in the animals resuscitated with HTS when compared with their NS counterparts, in blood (51.8 +/- 4.3 vs. 82.0 +/- 3.3 and 78.4 +/- 4.8, P = 0.005), lung (40.0 +/- 4.1 vs. 93.2 +/- 2.1 and 80.9 +/- 4.7, P = 0.002), and spleen (56.4 +/- 3.8 vs. 85.4 +/- 4.2 and 90.1 +/- 5.9, P = 0.05). Intracellular killing of bacteria increased markedly (P = 0.026) and superoxide generation was enhanced upon exposure to HTS (775.78 +/- 23.6 vs. 696.57 +/- 42.2, P = 0.017) despite inhibition of MAP kinase and NFkappaB activation. CONCLUSIONS: HTS significantly enhances intracellular killing of bacteria while attenuating receptor-mediated activation of proinflammatory cascades. PMID- 12894020 TI - Influence of modest endotoxemia on postoperative antithrombin deficiency and circulating secretory immunoglobulin a levels. AB - OBJECTIVE: To evaluate the influence of modest endotoxemia on postoperative antithrombin deficiency and cholestasis. SUMMARY BACKGROUND DATA: It has not been determined whether endotoxin translocation in small amounts is a physiological phenomenon or whether it is a potential health hazard. METHODS: Blood endotoxin, antithrombin III (ATIII), secretory immunoglobulin A (sIgA), which was selected as a marker of cholestasis, C-reactive protein (CRP), and alpha-1-antitrypsin (AAT) concentrations were measured from the 20 patients undergoing curative gastrectomy for gastric cancer preoperatively and postoperatively. Portal and systemic blood samples were taken for the analysis of endotoxin and interleukin-6 (IL-6) concentrations during surgery in these patients. RESULTS: Although plasma endotoxin levels showed a significant increase during surgery, we did not find a correlation with ATIII, sIgA, CRP, and IL-6 levels. Systemic blood endotoxin levels during surgery correlated with a postoperative rise of serum AAT levels. Plasma ATIII levels transiently decreased on the first and third postoperative day, and sIgA levels were shown to increase on the seventh postoperative day. There was a weak relationship between the extent of postoperative endotoxemia and a reduction in ATIII concentrations. CONCLUSIONS: The influence of modest endotoxemia on postoperative antithrombin deficiency and cholestasis was limited, and increased translocational endotoxemia during abdominal surgery may be a physiological phenomenon to trigger off an acute-phase protein response. PMID- 12894021 TI - Endothelin(A) receptor blockade reduces ischemia/reperfusion injury in pig pancreas transplantation. AB - OBJECTIVE: The effect of prophylactic administration of a selective endothelin(A) receptor antagonist (ET(A)-RA) on ischemia/reperfusion injury in an experimental model of graft pancreatitis after pancreas transplantation was evaluated. SUMMARY BACKGROUND DATA: It is well established that endothelin-1 (ET-1), a powerful vasoconstrictor, plays an important role in the development of pancreatitis. Recent studies have shown a beneficial effect of endothelin receptor antagonists in the therapy for experimental pancreatitis. METHODS: Relevant ischemia/reperfusion injury was induced in pig pancreas transplants after 6 hours hypothermic preservation in University of Wisconsin solution. The recipients were randomized into 2 groups: control pigs received isotonic saline and the treated group received the selective ET(A)-RA BSF 208075 at the beginning of reperfusion. On postoperative days 2 and 5, animals were relaparotomized to obtain tissue specimens. Blood monitoring included lipase, amylase, C-reactive protein, trypsinogen-activation peptide, thiobarbituric acid-reacting substances, and ET 1. Partial oxygen tension (p(ti)O(2)) was measured by a Clarke-type electrode and blood flow by laser doppler. A semiquantitative score index was used for assessment of histologic injury and for immunohistochemical analysis of ET-1 and ET(A) receptor expression. Tissue mRNA levels of prepro ET-1, ET(A) receptor, pro interleukin (IL)-6, and pro-IL-1beta were quantified using TaqMan real-time reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: Prophylactic treatment with ET(A)-RA significantly reduced the severity of graft pancreatitis evidenced by C-reactive protein. The finding of transient capillary perfusion at the beginning of reperfusion supports the application of the ET(A)-RA during this period. The dramatic increase of plasma ET-1 in the therapy group is a clear evidence of effective receptor blockade. Mean trypsinogen-activation peptide levels from the portal venous effluent, but not mean systemic plasma TAP values were significantly lower in the treated group. Analysis of p(ti)O(2) and blood flow revealed a significant improvement of capillary perfusion and blood flow in the treated group and was associated with relevant reduction of tissue injury. Intrapancreatic ET-1 and IL-6 mRNA expression and ET-1 protein levels were significantly lower in the therapy group as compared with the control group. In contrast, ET(A) mRNA showed a marked up-regulation by ET(A) receptor blockade. CONCLUSION: Application of a ET(A)-RA reduces ischemia/reperfusion induced graft pancreatitis in a pig transplantation model by improving microcirculation and reducing tissue injury. PMID- 12894022 TI - Right lobe living donor liver transplantation-addressing the middle hepatic vein controversy. AB - OBJECTIVE: To describe our approach in the decision-making for taking the middle hepatic vein with the graft or leaving it with the remnant liver in right lobe live donor liver transplantation. SUMMARY BACKGROUND DATA: Right lobe living donor liver transplantation has been successfully performed. However, the extent of donor hepatectomy is still a subject of debate and the main considerations in the decision making are graft functional adequacy and donor safety. METHODS: An algorithm based on donor-recipient body weight ratio, right lobe-to-recipient standard liver volume estimate, and donor hepatic venous anatomy was used to decide the extent of donor hepatectomy. This algorithm was applied in 25 living donor liver transplant operations performed between January 1999 and January 2002. In grafts taken without the middle hepatic vein, anterior segment tributaries draining into it were not reconstructed. Outcomes between right lobe liver transplants with (Group I) and without (Group II) the middle hepatic vein were compared. RESULTS: Ten grafts included the middle hepatic vein and 15 did not. The mean graft to recipient standard liver volume ratio was 58% and 64% in Groups I and II, respectively, and the difference was not statistically significant. Donors from both groups had comparable recovery, with 2 complications, 1 from each group, requiring a percutaneous drainage procedure. The recipient outcomes were, likewise, comparable and there was 1 case of structural outflow obstruction in Group I, which required venoangioplasty and stenting. There were 2 recipient mortalities, 1 due to a biliary complication and the other to recurrent hepatitis C. Another patient required retransplantation for secondary biliary cirrhosis. The overall actuarial graft and patient survival rates are 84% and 96%, respectively, at a median follow-up of 16 months. CONCLUSION: Based on certain preoperative criteria, a right lobe graft can be taken with or without the middle hepatic vein with equally successful outcomes in both the donors and recipients. The decision, therefore, of the extent of right lobe donor hepatectomy should be tailored to the particular conditions of each case. PMID- 12894023 TI - Aortoesophageal fistula: value of in situ aortic allograft replacement. AB - PURPOSE: The purpose of this report is to describe our experience in management of aortoesophageal fistulas (AEF) with special emphasis on the value of in situ aortic allograft replacement. PATIENTS: Nine patients presenting with AEF were observed between May 1988 and April 2002. There were 4 men and 5 women with a mean age of 54.3 years (range, 32-77 years). Six patients presented secondary AEF after aortic repair. Two patients presented primary AEF after rupture of an atherosclerotic aneurysm into the esophagus. In the remaining patient, AEF was caused by swallowing a fishbone. In 6 cases involving true AEF with a direct communication between the aorta and esophagus, massive exsanguinating hematemesis occurred. It was usually preceded by minor sentinel bleeding. Two patients presented esophagoparaprosthetic fistula (EPPF). One patient presented primary AEF that was contained by a large thrombus in the communication. The clinical picture in these 3 patients involved severe sepsis without hemorrhage. RESULTS: Two patients died as a result of massive hemorrhage before assessment and surgical treatment could be undertaken. One 77-year-old woman presenting EPPF refused to undergo surgery and died because of infection. The remaining 6 patients underwent surgical treatment with various outcomes. One man died during thoracotomy caused by exsanguinating hemorrhage. One woman presenting EPPF was treated by exclusion followed by ascending aorta to abdominal aorta bypass grafting, removal of the prosthesis, esophageal exclusion, and directed esophageal fistula. She died of infection. The other 4 patients were treated by in situ aortic allograft replacement. The damaged esophagus was repaired by using the Thal technique in 1 patient. In the remaining 3 cases subtotal esophagectomy was performed in association with cervical esophagostomy, ligation of the abdominal esophagus, gastrostomy, and jejunostomy. One patient died of sepsis during the first 24 hours after the operation. The other 3 patients underwent secondary esophagoplasty and survived with no further sign of infection. Mean duration of follow-up in the survivor group was 53 months (range, 15-95 months). Overall 6 patients, including 3 that did not undergo surgical treatment, died and 3 patients survived. CONCLUSION: Our experience confirms that AEF is a rare but catastrophic disorder. In situ allograft replacement usually in association with subtotal esophagectomy appears to be an excellent salvage modality whenever emergency surgery is feasible. PMID- 12894024 TI - Multiple Objective Measures of Skill (MOMS): a new approach to the assessment of technical ability in surgical trainees. AB - OBJECTIVE: The assessment of surgical technical skills has become an important topic in recent years. This study presents the validation of a 6-task skills examination for junior surgical trainees (at the level of the Membership of the Royal College of Surgeons). SUMMARY BACKGROUND DATA: Six tasks were evaluated in a project that also examined the feasibility of this method of assessment. The tasks were knowledge of sutures and instruments; knowledge of surgical devices; knot formation; skin-pad suturing, closure of an enterotomy; excision of a skin lesion; and laparoscopic manipulation. Comparisons were made between a group of junior trainees (n = 13), and a group of seniors (n = 8). RESULTS: Each of the 6 tasks was able to be used to discriminate between the 2 groups. In all, there were 19 primary analyses across the 6 tasks, and 17 of these showed significant differences between the groups (P values ranging from 0.037 to < 0.001). There was generally a strong correlation between the analyses, and when a mean rank was calculated, the difference between groups was significant (P = 0.005 on Mann Whitney U test; mean ranks 13.9 and 6.3 [of 21], for juniors and seniors respectively). Reliability of the 6-task assessment was very good at 0.70 (Cronbach's Alpha). CONCLUSIONS: A skills examination is a feasible and effective method of assessing the technical ability of basic surgical trainees. PMID- 12894025 TI - Repair of full-thickness defects in alimentary tract wall with patches of expanded polytetrafluoroethylene. PMID- 12894026 TI - Mitral anterior leaflet repair: ongoing validation. PMID- 12894027 TI - New methods of brain stimulation: what they tell us about the old methods and about the brain. PMID- 12894029 TI - Managing the risks of repetitive transcranial stimulation. PMID- 12894030 TI - Putative common pathways in therapeutic brain stimulation for affective disorders. AB - Brain stimulation methods in the treatment of affective disorder are electroconvulsive therapy (ECT), transcranial magnetic stimulation (TMS), vagus nerve stimulation (VNS), and deep brain stimulation (DBS). Clinically, ECT is considered the strongest remedy in the treatment of severe depression, especially depression with psychotic features, and in the elderly. TMS, despite positive reports, is somewhat more controversial. VNS has, so far, only been used in treatment-resistant depression with limited results. DBS may be of potential use in rare cases of treatment-resistant cases of affective disorder. This article highlights the similarities and differences between the four stimulation methods. The main difference is the seizures necessary in ECT. A stronger involvement of the hippocampus following experimental seizures compared with effects in that region induced by TMS and VNS might explain the consistent findings of the superiority of ECT in the most severe cases of affective disorder. PMID- 12894031 TI - Mechanisms and the current state of transcranial magnetic stimulation. AB - Transcranial magnetic stimulation (TMS) is unique among the current brain stimulation techniques because it is relatively non-invasive. TMS markedly differs from vagus nerve stimulation, deep brain stimulation and magnetic seizure therapy, all of which require either an implanted prosthesis or general anesthesia, or both. Since its rebirth in its modern form in 1985, TMS has already shown potential usefulness in at least three important domains-as a basic neuroscience research instrument, as a potential clinical diagnostic tool, and as a therapy for several different neuropsychiatric conditions. The TMS scientific literature has now expanded beyond what a single summary article can adequately cover. This review highlights several new developments in combining TMS with functional brain imaging, using TMS as a psychiatric therapy, potentially using TMS to enhance performance, and finally recent advances in the core technology of TMS. TMS' ability to non-invasively and focally stimulate the brain of an awake human is proving to be a most important development for neuroscience in general, and neuropsychiatry in particular. PMID- 12894032 TI - Beyond the treatment of epilepsy: new applications of vagus nerve stimulation in psychiatry. AB - Vagus nerve stimulation (VNS) in humans generally refers to stimulation of the left vagus nerve at the cervical level VNS is an established treatment largely devoid of severe side effect for medically refractory partial onset seizures and has been used in more than 16,000 patients. Over the past 5 years, applications in other neuropsychiatric disorders have been investigated with a special emphasis on depression. Recent data from an open-label, multi-center pilot study involving 60 patients suggest a potential clinical usefulness in the acute and maintenance treatment of drug-resistant depressive disorder. The perspective of VNS as along-term treatment with the advantage of assured compliance makes it an interesting technique to potentially treat drug-resistant depression. However, definite therapeutic effects of clinical significance remain to be confirmed in large placebo-controlled trial. Results of clinical pilot studies involving patients suffering from obesity and Alzheimer's disease indicate that VNS might induce weight loss and improve cognition. Besides its clinical usefulness, VNS can be used as a research tool, allowing neurophysiologic investigations of the parasympathetic system and its interactions with other parts of the central nervous system. PMID- 12894033 TI - Mechanisms and the current state of deep brain stimulation in neuropsychiatry. AB - Deep brain stimulation (DBS) is established as a therapy for movement disorders, and it is an investigational treatment in other neurologic conditions. DBS precisely targets neuroanatomical targets deep within the brain that are proposed to be centrally involved in the pathophysiology of some neuropsychiatric illnesses. DBS is nonablative, offering the advantages of reversibility and adjustability. This might permit therapeutic effectiveness to be enhanced or side effects to be minimized. Preclinical and clinical studies have shown effects of DBS locally, at the stimulation target, and at a distance, via actions on fibers of passage or across synapses. Although its mechanisms of action are not fully elucidated, several effects have been proposed to underlie the therapeutic effects of DBS in movement disorders, and potentially in other conditions as well. The mechanisms of action of DBS are the focus of active investigation in a number of clinical and preclinical laboratories. As in severe movement disorders, DBS may offer a degree of hope for patients with intractable neuropsychiatric illness. It is already clear that research intended to realize this potential will require a very considerable commitment of resources, energy, and time across disciplines including psychiatry, neurosurgery neurology, neuropsychology, bioengineering, and bioethics. These investigations should proceed cautiously. PMID- 12894034 TI - New developments in electroconvulsive therapy and magnetic seizure therapy. AB - New findings regarding the mechanisms of action of electro-convulsive therapy (ECT) have led to novel developments in treatment technique to further improve this highly effective treatment for major depression. These new approaches include novel placements, optimization of electrical stimulus parameters, and new methods for inducing more targeted seizures(eg, magnetic seizure therapy [MST]). MST is the use of transcranial magnetic stimulation to induce a seizure. Magnetic fields pass through tissue unimpeded, providing more control over the site and extent of stimulation than can be achieved with ECT. This enhanced control represents a means of focusing the treatment on target cortical structures thought to be essential to antidepressant response and reducing spread to medial temporal regions implicated in the cognitive side effects of ECT. MST is at an early stage of development. Preliminary results suggest that MST may have some advantages over ECT in terms of subjective side effects and acute cognitive functioning. Studies designed to address the antidepressant efficacy of MST are underway. As with all attempts to improve convulsive therapy technique, the clinical value of MST will need to be established through controlled clinical trials. This article reviews the experience to date with MST, and places this work in the broader context of other means of optimizing convulsive therapy in the treatment of depression. PMID- 12894035 TI - Obesity - a global problem. PMID- 12894036 TI - Factors affecting the early discharge of the indoor psychotic patients - a sociodemographic survey in a specialized institution. AB - This is descriptive type of cross-sectional study carried out among 100 discharged patient in a specialized psychiatric hospital. Socio-demographic study revealed that among 100 cases 97% were admitted with schizophrenia, 35% with bipolar affective disorder, 10% with psychotic depression, 5% with brief psychotic disorder, 2% with schizoaffective disorder, 1% with schizophreniform disorder. Their mean hospital stay was 19 days with highest mean hospital stay in schizoaffective disorder (32 days). Mean age of the patient was 26.5 years and highest no. of patients (52%) were in 20-25 years age group. Male female ratio of the patient was 3.761: 1. Occupational distribution of the cases family history of psychiatric illness in 1st degree and 2nd degree relatives was present and mean duration of illness was 73.4 months. Statistically significant independent variables that have impact on length of hospital stay (Dependent patient (p<0.007); electro convulsive therapy (p<0.005); family history of psychiatric illness (p< 0.046), Treatment with typical antipsychotic drugs (p <0.033); treatment with atypical antipsychotic drugs (p <0.000), Rx with combination of typical and atypical antipsychotic drugs (p<0.012); treatment with depot preparation (p<0.011); Treatment with combination of oral antipsychotic drugs (p<0.003) and patient hailing from rural area (p<0.026). PMID- 12894037 TI - A retrospective study on genito urinary Rhabdomyosarcoma. AB - A retrospective study was carried out on Genitourinary Rhabdomyosarcoma in the Department of Paediatric Surgery of Bangabandhu Sheikh Mujib Medical University, Dhaka, Bangladesh during the period January 1990 to December 1999. Total thirteen patients were treated during this period, age ranging from 1 month to 12 years. Eight patients were male and five were female (M: F = 1.6: 1). Diagnosis was made by history, thorough physical examination, digital rectal examination, ultrasonography, intravenous urography, CT scan and biopsy. All the patients were treated with chemoradiation theraphy as a first line of management and surgical excision was done for local residual tumours. Eight children (61.5%) were asymptomatic after completion of chemo-radiation therapy. Three (23.07%) patients discontinued the chemo-radiation schedule. Residual mass was present in one (7.7%) case for which partial cystectomy was done and the patient died in the 5th post operative day. Another patient (7.7%) who had pulmonary metastasis at the time of diagnosis, died during chemo-radiation therapy. PMID- 12894038 TI - Correlation between severity of coronary artery athesclerosis and duration and severity of diabetes mellitus in type 2 diabetic patients. AB - Hundred patients of Type 2 diabetes mellitus with coronary artery disease undergoing diagnostic coronary angiography were studied to determine the relation between the severity and duration of diabetes mellitus and the severity of coronary artery disease. A coronary scoring system using segmental distribution method was used to determine the severity of coronary artery disease and severity of diabetes was assessed by the level of fasting blood glucose within 48 hours of the procedure and also considering the types of treatment received for diabetes mellitus. No significant correlation was demonstrated between the severity of coronary artery disease and the severity (r =.089602; P>0.1) nor the duration (r =0.07865; P> 0.1) of diabetes mellitus on univariate analysis. So the Type 2 diabetic patients had an increased incidence of atherosclerosis in their coronary angiograms did not reflect an angiographically evident progressive coronary artery disease and results of this study suggest that the progress of large vessel coronary athesclerotic disease in diabetes mellitus of adult onset takes place independently of the progress of diabetes mellitus itself. PMID- 12894039 TI - Epidemiological characteristics of sixty five cases of Kala-azar attending to a laboratory in Mymensingh. AB - To find out the epidemiological characteristics of the patients with Kala-azar attending to a pathology laboratory at Mymensingh we studied retrospectively in a total of 65 patients. These patients were suspected to be Kala-azar as they were suffering from usually chronic fever, anaemia and splenomegaly. They were referred for serological detection of anti-k39 Leishmanial antibody by Immunochromatographic test (ICT) method. ICT positive cases were included for study. Majority cases were suffering from fever of more than 3 months (40%). Out of 65 patients 43 (66.15%) were males and 22 (33.85%) were females with a M:F ratio 2:1. Patients of 10 or less then 10 years of age were 17 (26.15%), in 11-20 years 23 (35.38%), in 21-30 years 13 (20.00%), in 31-40 years 5 (7.69%), in 41-50 years (6.15%) and in more than 50 years 3 (4.62%). Distribution of patients in different districts were: Mymensingh 54 (83.08%), Tangail 5 (7.69%), Jamalpur (4.62%), Netrokona 1 (1.54%) and Kishorgonj 1 (1.54%). In Mymensingh district patients were distributed as following Upazilla:Trishal 22 (40.74%), Fulbaria 11 (20.37%), Mymensingh Sadar 7 (12.96%), Gaforgaon 7 (12.96%),Bhaluka 5 (9.96%), Fulpur 1 (1.85%) and Nandail 1 (1.85%). Patients were found to scattered among different Unions of the affected Upazillas. The details addresses of the Kala azar patients are recorded in authors computer databases. This may help in the further study regarding pathogenesis, reservoir and vectors of Leishmania in endemic areas. It may also helps in the community based study of Kala-azar. Present study supports that Kala-azar is still prevailing in the south-western region of greater Mymensingh in Bangladesh of which males and the adolescents are more affected. PMID- 12894040 TI - Immunochromatographic (rK39) strip test in the diagnosis of visceral leishmaniasis in Bangladesh. AB - Immunochromatographic strip test (ICT strip test) for the diagnosis of visceral leishmaniasis was evaluated in this study in the context of a case-control study. A total sixty consecutive cases of kala-azar admitted in all four Medicine Units of Mymensingh Medical College Hospital during the period of May 2002 to February 2003 was included here. Parasitological confirmation was done by demonstration of leishmania donovani bodies in bone marrow or splenic aspiration in all cases. A total 120 controls was taken of which sixty were asymptomatic endemic controls with no previous history of kala-azar and sixty were admitted patients suffering from diseases other than kala-azar (malaria, tuberculosis, enteric fever and chronic liver disease). ICT strip test for kala-azar was done in all cases and controls. Only 2 of the confirmed kala-azar cases were negative and the remaining 58 cases were positive for ICT strip test which gives the sensitivity of this test 96.6%. Among the controls, 118 were negative for ICT strip test and two of the asymptomatic controls were positive for this test with no clinical evidence of kala-azar. So, the estimated specificity of ICT strip test is 98.3%. The predictive value for a negative result was 98.3% and for a positive result was 96.6%. The ICT strip test is easy, quick, requires no technical facilities with higher sensitivity and specificity entails it to be the ideal test for the diagnosis of kala-azar in field level. PMID- 12894041 TI - Pattern of aerobic bacteria with their drug susceptibility of surgical inpatients. AB - The pattern of aerobic bacteria with their antibiotic susceptibility isolated from infected patients in one of the surgical units at Mymensingh Medical College Hospital during the period from September to November' 2000 were reported in the present study. Out of 74 clinical samples, 52 were wound swabs, 18 were pus and 4 were urine. Bacterial growth was yielded in 43 samples and the distribution of isolates was as follows: Pseudomonas spp 16, Esch.coli 13, Staphylococcus aureus 08, Klebsiella spp. 03 and others 03. Majority (61.5 %) of culture positive results were found in wound swabs. Antibiotic susceptibility pattern showed considerable variation within Gram negative bacterial isolates. All the Esch.coli and Klebsiella isolates were resistant to amoxicillin. Ceftriaxone (65.6% and 100% respectively) and ciprofloxacin (71.4% and 100%) still appeared to be highly sensitive for both species. Over 93% strains of Pseudomonas were sensitive to Ceftazidime and aztreonam. Whereas, over 43% of same strains were resistant to ciprofloxacin. Over 87% strains of Staph.aureus were resistant to penicillin but sensitive to erythromycin. Whereas, 100% of those strains were sensitive to cloxacillin. Over 50% of all isolates were sensitive to gentamicin but resistant to cefalexin and cotrimoxazole. It was suggested to be careful regarding selection of antibiotic regime in surgical cases to minimize incoming higher magnitude of drug resistance among bacteria in near future. PMID- 12894042 TI - Pattern of bacteria and their antimicrobial susceptibility isolated from inanimate objects and hospital personnel. AB - Different inanimate samples collected from surgery wards and general operation theatre (GOT) as well as animate samples from surgery personnel were subjected to aerobic culture with the aim of identifying source of infections in surgical cases. The pattern of aerobic bacteria with antibiotic susceptibility isolated from those samples were reported in the present study. Out of 91 such samples, 53 and 26 were from different locations in general wards and OT respectively and 12 were comprising of hand swabs, throat swabs and nasal swabs from surgery personnel. Pathogenic bacteria were isolated from 34 samples having Pseudomonas spp. (18/34) in the top of the list followed by Esch. coli (14). Among the Esch.coli isolates resistance to cotrimoxazole, amoxicillin and doxycycline were 78.7%, 71.4% and 57.1% respectively, whereas sensitivity to aztreonam, ceftazidime, gentamicin and ciprofloxacin were 100%, 85.8%, 78.7% and 71.4% respectively. Ceftriaxone yielded intermediate sensitive results in 21.4% strains. Isolates of Pseudomonas spp. also showed very high sensitive results to aztreonam, ceftazidime and ceftriaxone (94.7%, 94.7% and 84.2% respectively). Ciprofloxacin and gentamicin were sensitive in 68.4% and 57.9% respectively. Susceptibility pattern showed well correlation with clinical response in infected cases. PMID- 12894043 TI - A prospective study on histochemical observation of thyroid gland at prepubertal Black Bengal goat. AB - In the present study, Period Acid Schiff (PAS), Verhoeff's and Van Gieson stains were used in the Department of Anatomy and Histology to observe the histochemical architecture of thyroid gland at prepubertal Black Bengal goat. Thyroid follicular cells were cuboidal in prepubertal goats. These cells showed moderate PAS-positive reaction in prepubertal group. PAS activity in the basement membrane of the thyroid follicles was mild in prepubertal Black Bengal goats. The lumen of the thyroid follicles contained homogenous intense PAS positive colloid materials in prepubertal goats. Parafollicular cells showed intense PAS positive reaction for glycogen in prepubertal goats. Capsule, connective tissue septa, interfollicular connective tissue of the thyroid gland mainly composed of collagen fiber. The wall of blood vessels in the capsule and interfollicular blood capillaries was composed of collagen fiber and elastic fiber. Elastic fiber formed the inner layer of the blood vessels showed blue black to black color. Inner layer of the capsule penetrated the gland parenchyma divided the gland into indistinct lobe and lobules. Collagen fiber distributed around the thyroid follicle formed the interfollicular stroma of the thyroid gland. The results of the present study can also be used to compare with that of human specially in AutoImmune Thyroiditis (AIT). PMID- 12894044 TI - Gall stone in pregnancy. AB - Gallbladder disease is four times as common in women as in men, and pregnancy appears to contribute to the development of gallstones. During pregnancy, most women receive ultrasound scans, which are highly sensitive in detecting gallstones. The study was undertaken to evaluate the prevalence of gallstones among the pregnant woman. The aim was to determine any significant difference in the prevalence of gallstone among the pregnant woman compared to the generalized prevalence of gallstone in our community. METHODS: The maternal gallbladder was examined in 1336 consecutive obstetric ultrasound scans performed for medical indications at 3rd trimester of pregnancy. The study was conducted in Centre for Nuclear Medicine and Ultrasond (CNMU) Mymensingh on randomly selected subjects, among those who came here for ultrasound evaluation of pregnancy. 500 control were taken from age group between 20-30 yrs who came here for ultrasound examination of lower abdomen other than pregnancy. RESULTS: Gallstone were found in total one hundred eight cases. Prevalence of gallstone in pregnant woman was 8.08% (n=108). The age ranges were 20 to 45 yrs with mean age of 28.21 +/- 4.95 (mean +/- SD) yrs. The highest prevalence (12.71%) were seen in 3rd or higher gravida group with mean age of 30.32 +/- 4.74 yrs. Significantly higher cases of gallstone were found in multiparity compared to control population of same age group. CONCLUSIONS. Repeated pregnancy causes increased gallstone formation due to changes in gallbladder kinetics leading to stasis and stone formation. PMID- 12894045 TI - Experience of cranial ultrasonography in admitted symptomatic neonates. AB - In infants who have experienced perinatal injury or who have neurological symptoms, it is often difficult on the basis of history and clinical examination to come to a proper diagnosis. Cranial ultrasonography of the symptomatic neonates can often diagnose the underlying pathology. Cranial ultrasonography of 28 admitted neonates with neurological symptoms was done at Community Based Medical College Hospital (CBMCH), Mymensingh. In 11 cases (39%) there were positive findings in ultrasonography. Of them, 7 (25%) were Hypoxic ischaemic encephalopathy (HIE) and 4 (14%) had dilated ventricles. No abnormal ultrasonography findings were noted in the remaining neonates. Before closure of the fontanelles, Ultrasonography can help as a cheaper, easier and acceptable method to diagnose some neurologic problems in children. PMID- 12894046 TI - Antioxidant vitamins improves hemoglobin level in children with group a beta hemolytic streptococcal infection. AB - A study was done on school children infected with group A beta hemolytic streptococci to examine whether antioxidant vitamins play a role in improving the hemoglobin level. A total of 606 primary school children aged 5 to 15 years were randomly divided into two intervention groups. Group 1 (n=299) was treated with pehnoxymethyl penicillin V and group 2 (n=307) was treated with phenoxymethyl penicillin V plus antioxidant vitamins for eight weeks. From each group two blood samples were drawn in acute and convalescent (after eight weeks) states. Before treatment, mean hemoglobin values were 11.0 and 10.8 mg/dL in groups 1 and 2 respectively. After treatment hemoglobin values were 10.5 and 11.6 mg/dL respectively. Values were significantly decreased in group 1 (P=0.0001), whereas increased in group 2 (P=0.001). Adjustment for age and sex by ANCOVA confirmed the difference in hemoglobin levels between group (LS means-0.5 vs 0.8 in groups 1 and 2 respectively (P=0.0001). Hemoglobin level increases after antioxidant vitamin supplementation in children suffering from group A beta hemolytic streptococcal infection. PMID- 12894047 TI - Upper gastrointestinal endoscopy in children - an experience at a paediatric gastroenterology unit. AB - Upper gastrointestinal endoscopy has changed the management of upper gastrointestinal problems in children. The aim of this communication is to share our experience with 153 cases on whom upper gastrointestinal endoscopy was done over a period of 24 months at a paediatric gastroenterology unit of a tertiary care hospital of Dhaka, Bangladesh. Children who attended the department with various gastrointestinal problems are the subjects of this paper. Intravenous midazolam and 10% pharyngeal xylocain were used in majority of cases for sedating the children. The ages of the children were between 15 months to 15 years (9.41+/ 3.22 years). The positive diagnostic yield was 92 out of 153 cases (60.1%). The major indication for doing endoscopy in the present series was recurrent abdominal pain (51.6%), followed by upper gastrointestinal bleeding (28.8%). Combining histopathological findings and CLO/rapid urease tests the overall positive yield of recurrent abdominal pain was 45 out of 79 (57%). The sources of upper gastrointestinal bleeding could be identified in 79.5% cases. Esophageal varices indicating portal hypertension were found in 62.5% children who were endoscoped for unexplained splenomegaly with or without ascitis. Endoscopy has become a safe and valuable procedure in the management of upper gastrointestinal problems in children and gastric antral biopsy has increased the positive diagnostic yield of recurrent abdominal pain in the studied children. PMID- 12894048 TI - Evaluation of hospitalized infants and young children with bronchiolitis - a multi centre study. AB - Four hundred and twenty nine young children with bronchiolitis admitted consecutively in different hospitals of Bangladesh were evaluated. Three hundred and forty eight children studied for their putative risk factors, clinical profile, management and the outcome. Both cases and controls were examined for respiratory syncytial virus (RSV) antibody status. The diagnosis of bronchiolitis was made on the basis of first attack of wheeze in previously healthy children below two years of age. Detailed history including the possible risk factors, the management and daily follow-up on the ward and the outcome at discharge were documented through a structured questionnaire. Chest x-ray was done in each case to find out the radiological changes. Blood of 266 patients and 30 controls were studied for RSV IgM and IgG antibody by ELISA. There were 66% male and 34% female children. The median age of the children was 3.0 months and 82.7% were below 6 months of age. Most of the babies were born term (88%), with ABW (73%), by normal vaginal delivery (88%). Exclusive or predominant breast-feeding were given in 72% cases. The location of the patient was rural in 55% cases. Around half of the parents were illiterate or slightly educated (up to 5 years schooling) fathers 46.5% and mothers 56% and majority of the parents were poor (74%). In 52% cases the number of family members in one room were four or more. Half of the parents (52%) were smokes and there was atopy in 26.5% families. The clinical features of bronchiolitis were mostly cough (99%), respiratory distress (97%), feeding difficulty (93%) and fast breathing (96%) (median RR 68/min). Fever (1000F or more) was in only 33% cases, though parents complained in 90% cases. All children (100%) had wheeze and crackles in lungs in 96% cases. Liver could be palpable in 83% and spleen in 42% cases. Important radiological features were increased translucency (96%), increased interstitial markings (87%), hyperinflation (75%) and streaky densities (61%). In 69.6% cases TLC was 12,000 or less and only 15% with a neutrophil fraction greater than 60%. Children were positive for IgM antibody in 43.6% cases and both IgM and IgG in 5.3% cases. The main modalities of treatment were antibiotics (99%) (Ampicillin, 76%), oxygen therapy (83%), nebulised salbutamol (76%) and intravenous fluid (51%). The median duration of hospital stay was 4 days. Most of the children were discharged with improvement (96%) with 2% mortality. Not a single case was diagnosed as bronchiolitis in hospitals outside Dhaka. Cefrtiaxone (72.5%) and parenteral steroids (70.5%) were the mainstay of therapy there. PMID- 12894049 TI - Hairy cell leukaemia with advanced treatment - a case report. AB - A case of hairy cell leukaemia (HCL), a rare leukaemia, is reported here. The patient was presented with high grade continuous fever with left upper abdominal discomfort for 6 days. He was moderately anaemic, had no peripheral lymphadenopathy with mild hepatosplenomegaly. He was anaemic (Hb-7.8 gm/dl), total leukocyte count was 20 x 109/L. Peripheral blood film showed lymphocytosis (92%) with neutropenia (8%) and absolute neutophil count (ANC) was 1 x 109/L. On review, 88% of the peripheral cells had peripheral hairy projections resembling hairy cell (HC). Bone marrow examination was consistent with HCL (morrow hairy cell = 52%) including marker studies. Tartrate resistant acid phosphatase test (TRAP) was also positive. He had opportunistic mycobecterial infection giving a positive bronchial lavage for acid fast bacilli. After controlling the infection he was advised a single dose chemotherapy of 2-chlorodeoxyadenosine (2-CDA). After that he was in partial remission and after 25 months clinical and pathological relapses occurred and a second dose of 2-CDA was given and the patient went into complete remission. PMID- 12894050 TI - Hydrops fetalis: report of 3 cases. AB - Ultrasound is a well recognized imaging modality for prenatal diagnosis of fetal congenital anomalies. Reported cases are hydrops fetalis diagnosed during ultrasonic evaluation of fetal condition in the Center for Nuclear Medicine and ultrasound (CNMU), Mymensingh. Only fetal ascites was detected in one case of 26 +/- 2 wks of gestation, fetal ascites with hydramnios and thick placenta was seen in another case of 28 +/- 2 wks of gestation. In third case, there was fetal ascites, scalp edema, hydrothorax and myelomeningocele with oligohydramnios at 20 +/- 2 wks of gestation; sonographic diagnosis was hydrops fetalis with myelomeningocele. Follow up was advised in first two cases and third case was terminated electively. To decrease the mortality rate and to improve the outcome of hydrops fetalis cases appropriate prenatal investigations and therapy is needed. Recent advances in prenatal ultrasound have made possible the early detection of hydrops fetalis which is helpful for proper management in time. PMID- 12894051 TI - A case report on Wilson's disease. AB - A case of Wilson's disease, a rare autosomal recessive disorder of copper metabolism is reported here. The patient was presented with the difficulty in speech and writing for 4 years and also on walking for 1 year. He also noticed difficulty to perform any work by hands for 6 months. He had splenomegaly and bilateral gynaecomastia. His speech was low volume slurred and monotonous, muscle tone was mildly increased, and gait was limping. Slit lamp examination of eye revealed bilateral Kayser-Fleischer ring with normal visual acuity. Investigations revealed low serum albumin(26 gram/L), increased alanine trans aminase ( A.L.T=57 U/L). Ultrasonogram of hepatobiliary system revealed coarse hepatic tissue echotexture with splenomegaly. Liver scan showed slightly nonuniform radiotracer distribution in the liver, there was slight increased bony uptake. Serum caeruloplasmin level was 11.51 mg/dl. 24 hours urinary copper excretion was 150 microgram per day. Liver biopsy revealed cirrhotic change. Now he was advised for taking copper chelating agent (penicillamine) in a dose of 1 gram/day. PMID- 12894052 TI - Thyroid status of normal pregnant woman. PMID- 12894055 TI - Injury mortality among American Indian and Alaska Native children and youth- United States, 1989-1998. AB - Injuries account for 75% of all deaths among American Indian and Alaska Native (AI/AN) children and youth, and AI/ANs have an overall injury-related death rate that is twice the U.S. rate for all racial/ethnic populations. However, rate disparities vary by area and by cause. To help focus prevention efforts, CDC analyzed injury mortality data by Indian Health Service (IHS) administrative area and by race/ethnicity. This report summarizes the results of these analyses, which indicate that although death rates for some causes (e.g. drowning and fire) have shown substantial improvement over time, rates for other causes have increased or remained unchanged (e.g., homicide and suicide, respectively). Prevention strategies should focus on the leading causes of injury-related death in each AI/AN community, such as motor-vehicle crashes, suicides, and violence. PMID- 12894056 TI - Diabetes prevalence among American Indians and Alaska Natives and the overall population--United States, 1994-2002. AB - Diabetes affects American Indians/Alaska Natives (AI/ANs) disproportionately compared with other racial/ethnic populations and has been increasing in prevalence in AI/AN populations during the past 16 years. To examine trends in diabetes prevalence among AI/ANs and the overall U.S. population and to describe disparities among these two populations, CDC analyzed data from the Indian Health Service (IHS) and the Behavioral Risk Factor Surveillance System (BRFSS). This report summarizes the results of that analysis, which indicate that diabetes continues to affect AI/ANs disproportionately and is becoming more common among younger populations. To combat this epidemic, knowledge and interventions from clinical trials and best-practice models should be translated to community-based prevention programs within AI/AN communities. PMID- 12894057 TI - Cancer mortality among American Indians and Alaska Natives--United States, 1994 1998. AB - In the United States, public health interventions to control infectious diseases, lower infant and maternal mortality, and improve basic sanitation have led to a substantial increase in life expectancy for American Indians and Alaska Natives (AI/ANs). During 1940-1995, average life expectancy among AI/ANs increased 39%, from 51 years in 1940 to 71 years in 1995; however, AI/ANs experienced a parallel increase in mortality rates for chronic diseases, including cancer, which is the second leading cause of death for AI/ANs nationally and the leading cause of death among Alaska Natives. A previous study examining cancer mortality rates during 1989-1993 documented lower cancer mortality rates for AI/ANs than for the overall U.S. population, with regional variation. To understand cancer mortality among AI/ANs subsequent to that period, the Indian Health Service (IHS) and CDC analyzed death certificate data provided by CDC's National Center for Health Statistics for deaths among AI/ANs in five U.S. geographic regions during 1994 1998. This report summarizes the results of that analysis, which indicate that cancer mortality rates among AI/ANs nationally were lower than cancer mortality rates for all U.S. racial/ethnic populations combined. Rates for AI/ANs varied by region, with the highest rates found in the Alaska and the Northern Plains regions. Plans or modifications for cancer prevention and treatment programs should account for regional variation, and programs to discourage smoking initiation, encourage tobacco cessation, and promote colorectal cancer screening among AI/ANs in the Alaska and the Northern Plains regions should be expanded. PMID- 12894058 TI - Bronchiolitis-associated outpatient visits and hospitalizations among American Indian and Alaska Native children--United States, 1990-2000. AB - Respiratory syncytial virus (RSV) is the most common cause of lower respiratory tract infection (LRTI) in young children worldwide. Approximately half of all LRTI-associated hospitalizations are caused by bronchiolitis, with RSV accounting for 50%-80% of all bronchiolitis cases. Bronchiolitis is an infection of the bronchial and bronchiolar epithelial cells, with subsequent inflammation and edema resulting in airway obstruction. This process manifests clinically as cough, wheezing, tachypnea, and respiratory distress. Because of the association between bronchiolitis and RSV infection, bronchiolitis is a good indicator of RSV disease; therefore, prevention strategies for RSV should reduce the rate of bronchiolitis. Rates of bronchiolitis-associated hospitalization for American Indian/Alaska Native (AI/AN) children are approximately twice that for the general population of U.S. children. This report describes the first estimate of rates of outpatient bronchiolitis-associated visits and updates rates of bronchiolitis-associated hospitalizations in these populations. Rates of bronchiolitis-associated outpatient visits and hospitalizations were higher for AI/AN children than for other U.S. children, and hospitalization rates for both groups increased during 1990-2000. This report underscores the high burden of bronchiolitis and the need for effective prevention programs for AI/AN communities. PMID- 12894059 TI - Vaccination coverage levels among Alaska Native children aged 19-35 months- National Immunization Survey, United States, 2000-2001. AB - In 2000, a total of 118,846 persons indicated that their race/ethnicity was Alaska Native (AN), either alone or in combination with one or more other racial/ethnic groups. AN groups comprise 19% of the population of Alaska and 0.4% of the total U.S. population. The AN grouping includes Eskimos, Aleuts, and Alaska Indians (members of the Alaska Athabaskan, Tlingit, Haida, or other AN tribes). Eskimo represented the largest AN tribal grouping, followed by Tlingit/Haida, Alaska Athabascan, and Aleut. Vaccination coverage levels among AN children have not been reported previously. This report presents data from the National Immunization Survey (NIS) for 2000-2001, which indicate that vaccination coverage levels among AN children aged 19-35 months exceeded the national health objective for 2010 (objective no. 14-22) for the majority of vaccines. This achievement indicates the effectiveness of using multiple strategies to increase vaccination coverage. Similar efforts might increase vaccination coverage in other rural regions with American Indian (AI)/AN populations. PMID- 12894062 TI - Oral retinoid therapy for dermatologic conditions in children and adolescents. AB - The efficacy of systemic retinoid therapy in a number of dermatologic diseases is well established; however, concerns about potential side effects limit their use, especially in children. We review the efficacy and toxicity of oral retinoids in the pediatric population. The acute mucocutaneous toxicities commonly observed are typically well tolerated, readily treatable, and reversible. Systemic toxicities include teratogenicity and effects on the musculoskeletal, neurologic, and gastrointestinal systems. Children, like adults, generally tolerate short term retinoid therapy without major complications. Concerns regarding serious systemic side effects are greater for those on high doses of oral synthetic retinoids for longer periods of time. Close patient monitoring and patient education can minimize the occurrence of complications. (J Am Acad Dermatol 2003;49:171-82.)Learning objective At the conclusion of this learning activity, participants should be familiar with use of oral retinoids for childhood dermatologic conditions such as psoriasis, acne, and ichthyoses as well as safety and risks associated with oral retinoid use in children and adolescents. PMID- 12894063 TI - Superficial fungal infections in 102 renal transplant recipients: a case-control study. AB - BACKGROUND: Renal transplant recipients are predisposed to superficial fungal infections caused by graft-preserving immunosuppressive therapy. Reports have documented a wide range of prevalence rates for superficial fungal infections in this patient group. OBJECTIVE: The aim of this study was to determine the prevalence and clinical and mycological features of superficial fungal infections in renal transplant recipients at our center. METHODS: One hundred two consecutively registered renal transplant recipients (34 women, 68 men) and 88 healthy age- and sex-matched persons acting as controls (30 women, 58 men) underwent screening for the presence of superficial fungal infection. Skin scrapings and swabs were obtained from the dorsum of the tongue, upper part of the back, toe webs, and any suspicious lesions. Nail clippings were also collected. All samples were examined by direct microscopy and were stained with calcofluor white. The samples were cultured in Sabouraud dextrose agar, mycobiotic agar, and dermatophyte test medium. Candida species were identified on the basis of germ-tube production, spore formation in cornmeal agar, and results of biochemical testing. Dermatophytes were identified on the basis of colonial and microscopic morphologic features in conjunction with results of physiologic evaluation (in vitro hair perforation test, urease activity, temperature tolerance test, and nutritional test). RESULTS: Sixty-five (63.7%) of the 102 renal transplant recipients had cutaneous-oral candidiasis, dermatophytosis, or pityriasis versicolor, whereas only 27 (30.7%) of controls had fungal infection. Pityriasis versicolor was the most common fungal infection in the patient group (36.3%), followed by cutaneous-oral candidiasis (25.5%), onychomycosis (12.7%), and fungal toe-web infection (11.8%). Pityriasis versicolor and oral candidiasis were significantly more common among the renal transplant recipients, whereas the frequency of dermatophytosis in patients and controls was similar. Candida albicans was the main agent responsible for oral candidiasis, and Trichophyton rubrum was the most common dermatophyte isolated. Analysis showed that age, sex, and duration of immunosuppression did not significantly affect the prevalence of superficial fungal infection. Cyclosporine treatment and azathioprine therapy were identified as independent risk factors for superficial fungal disease. CONCLUSIONS: The prevalence of opportunistic infections with Pityrosporum ovale and C albicans is increased among renal transplant recipients, probably owing to the immunosuppressed state of this patient population. However, renal transplant recipients are not at increased risk of dermatophytosis. PMID- 12894064 TI - Comparison of diagnostic methods in the evaluation of onychomycosis. AB - BACKGROUND: Onychomycosis is a common problem seen in clinical practice. Given the differential diagnosis of dystrophic nails, it is helpful to obtain a definitive diagnosis of dermatophyte infection before the initiation of antifungal therapy. Potassium hydroxide (KOH) preparation and fungal culture, which are typically used in the diagnosis of these infections, often yield false negative results. Recent reports have suggested that nail plate biopsy using periodic acid-Schiff (PAS) (Bx/PAS) stain may be a very sensitive technique for the diagnosis of onychomycosis. OBJECTIVE: The purpose of this study was to compare KOH preparation, culture, Bx/PAS stain, and calcofluor white (CW) stain in the diagnosis of onychomycosis and to determine their sensitivity and specificity. METHODS: We evaluated 105 patients with suspected onychomycosis using 4 diagnostic methods: KOH preparation, culture, Bx/PAS, and CW stain. CW stain binds to cellulose and chitin, and fluoresces when exposed to UV radiation. It is a highly sensitive and specific technique for the detection of dermatophytes. To determine the clinical usefulness and performance characteristics of each test, CW was chosen as the gold standard for statistical analysis. RESULTS: Of the patients, 93 had at least 1 of the 4 diagnostic methods positive for the presence of organisms. The following were calculated for each test: sensitivity; specificity; positive predictive value; and negative predictive value. The sensitivities of each of the techniques were as follows: KOH 80%; Bx/PAS 92%; and culture 59%. Both KOH and Bx/PAS methods were more sensitive than culture (P =.00002). Bx/PAS was also more sensitive than KOH (P =.03). The specificities were as follows: KOH 72%; Bx/PAS 72%; and culture 82%. The positive predictive value calculated for the different techniques were: KOH 88%; Bx/PAS 89.7%; and culture 90%. In terms of negative predictive value, the results were: KOH 58%; Bx/PAS 77%; and culture 43%. CONCLUSION: Bx/PAS is the most sensitive method for the diagnosis of onychomycosis. It is also superior to the other methods in its negative predictive value. It is indicated if other methods are negative and clinical suspicion is high, and potentially is the single method of choice for the evaluation of onychomycosis. PMID- 12894065 TI - Predisposing factors and clinical features of eczema herpeticum: a retrospective analysis of 100 cases. AB - BACKGROUND: Eczema herpeticum (EH) is a widespread herpes simplex virus infection of inflamed skin, most often occurring in patients with atopic dermatitis (AD). A monomorphic eruption of dome-shaped blisters and pustules in the eczematous lesions along with severe systemic illness lead to the clinical diagnosis, but atypical variants with disseminated slits may also occur. Topical use of corticosteroids is alleged to be a pathogenetic factor for EH, but predisposing factors for EH are largely unknown. Objective and methods We sought to characterize the clinical features and predisposing factors for EH. A retrospective analysis of 100 patients with EH seen from 1980 through 1996 and of 105 control patients with AD was performed. RESULTS: Fever and lymphopenia were associated with EH, whereas an increased erythrocyte sedimentation rate was frequently seen in patients with EH and control patients who were impetiginized. In 100 patients with EH, primary herpes simplex virus infection was likely in 20 patients, and a secondary herpes simplex virus infection was suggestive in 26 patients. In all, 13 patients had a second EH, whereas 3 patients had a third EH. Patients with EH had a significantly earlier onset of AD and a significantly higher total serum IgE level than the control patients. More than 75% of the patients with EH had not received corticosteroid treatment in the 4 weeks before onset of EH. CONCLUSIONS: The characteristics of patients with EH are those associated with severe manifestations of AD. The majority of EH occurs in patients with untreated AD, arguing against a role for topical corticosteroids in the development of EH. PMID- 12894066 TI - Nail Psoriasis Severity Index: a useful tool for evaluation of nail psoriasis. AB - The Nail Psoriasis Severity Index (NAPSI) is a numeric, reproducible, objective, simple tool for evaluation of nail psoriasis. This scale is used to evaluate the severity of nail bed psoriasis and nail matrix psoriasis by area of involvement in the nail unit. The NAPSI will be useful during clinical trials for evaluating response to treatment of psoriatic nails. The scale is reproducible, and because there are few data points, statistical analysis is simplified. PMID- 12894067 TI - Pharmacologic denervation of frown muscles enhances baseline expression of happiness and decreases baseline expression of anger, sadness, and fear. AB - BACKGROUND: Individuals with intense frowning are commonly perceived as expressing negative emotions. Anger, fear, and sadness are associated with corrugator ("frown") muscle activity. OBJECTIVE: We sought to study how faces were perceived by others after denervation of frown muscles with localized botulinum toxin injections for treatment of facial frown lines. METHODS: Facial photographs were taken from volunteers before and after botulinum toxin injection. These photographs were shown to viewers who were naive to the procedure and asked to rate the expressed intensity of anger, sadness, fear, and happiness. As reference for this task we used a standard set of pictures of facial affect displaying different intensity levels for each emotion tested. RESULTS: Of 40 viewers, 39 were able to discriminate different intensity levels (0%, 25%, 50%, 75%, and 100%) of emotional affect in the control task. According to their ratings faces with denervated frown muscle activity expressed relatively less anger (-40%), fear (-49%), sadness (-10%), and more happiness (+71%). CONCLUSION: Frown muscle activity is essential for both negative and positive emotional expressions. Temporary denervation using botulinum toxin enhances the facial expression of positive emotion resulting in a shift rather than a loss of facial affect. PMID- 12894068 TI - Achilles tendinitis in psoriasis: clinical and sonographic findings. AB - BACKGROUND: Involvement of the Achilles tendon is frequent in psoriatic arthritis, but it is easily missed at clinical examination. OBJECTIVE: To seek evidence of Achilles tendon abnormalities by means of sonography in psoriatic patients and to correlate sonographic findings with clinical symptoms (tendon and soft-tissue swelling, pain, and difficulty in walking). METHODS: Fifty-nine patients with plaque-type psoriasis (Psoriasis Area and Severity Index score, 3.7 34.7) and 50 healthy, aged-matched volunteers underwent clinical and sonographic evaluation of Achilles tendons and peritendinous structures. RESULTS: Eighteen (30.5%) of the 59 patients had clinical symptoms of Achilles tendinitis. Thirty five (59.3%) of the patients had sonographic abnormalities. Of these, 13 patients had clinically symptomatic abnormalities, and 11 had psoriatic arthritis. Degenerative tendinitis was the most frequent sonographic finding (76.9%) among patients with symptomatic conditions. Five patients with symptoms did not have sonographic alterations. None of the controls had clinical or sonographic changes. CONCLUSIONS: In psoriatic patients Achilles tendon abnormalities cannot be excluded even when they are clinically absent. PMID- 12894069 TI - Large B-cell lymphoma of the leg: clinical and pathologic characteristics in a North American series. AB - BACKGROUND: Large B-cell lymphoma (LBCL) of the leg is an uncommon subset of primary cutaneous B-cell lymphoma that has been described in a series of European patients. OBJECTIVE: Our purpose was to evaluate the clinical manifestation, diagnostic histopathology, immunophenotype, clinical course, and response to treatment of LBCL of the leg. METHODS: We conducted a retrospective case series of 3 patients with primary LBCL of the leg. RESULTS: The 3 elderly patients presented with progressive erythematous nodules on bilateral or unilateral lower extremities. All 3 patients had pre-existing peripheral edema or peripheral vascular disease. Histopathologic examination of the nodules showed dense lymphocytic infiltrates composed predominantly of large dysplastic lymphocytes that marked as B cells (CD20(+)). In 2 cases, the neoplastic cells were BCL-2 positive. All patients responded to initial therapy with localized electron beam radiation and chemotherapy but had disease progression. One patient had a complete and durable second response to anti-CD20 monoclonal antibody (rituximab). CONCLUSIONS: The patients described have similar clinical and histopathologic features to those previously described. There may be an association between LBCL and pre-existing lower-extremity vascular disease. Treatment of LBCL is difficult, but 1 patient responded well to systemic anti CD20 monoclonal antibody. PMID- 12894070 TI - The expression of insulin-like growth factor 1 in follicular dermal papillae correlates with therapeutic efficacy of finasteride in androgenetic alopecia. AB - BACKGROUND: It is generally believed that dihydrotestosterone is one of the pivotal mediators of hair loss in androgenetic alopecia (AGA). Finasteride, which blocks the conversion of testosterone to dihydrotestosterone, has now become an integral part of the current treatment approaches for male AGA. Several lines of evidence support the notion that dermal papilla (DP) cells represent the androgen target within the hair follicle. The specific molecular regulators modulated by androgens within hair follicles in the balding scalp are unknown. OBJECTIVE: The purpose of this study was to identify and quantify changes in expression of specific molecular hair growth regulators in DP of men with AGA treated with finasteride and correlate these findings to clinical efficacy. METHODS: Biopsy specimens were collected from 9 male patients from both the balding area and nonbalding occipital area before and after 4 months of finasteride therapy. DP were microdissected and total RNA was extracted from an equal number of DP from each biopsy specimen. The expression of various cytokines, including insulin-like growth factor (IGF)-1, was determined by reverse transcription polymerase chain reaction. The signals were detected by autoradiography. All 9 patients were given finasteride for 1 year and evaluated for efficacy at month 12. Efficacy was graded on a 7-point scale on the basis of comparison with initial baseline photography. RESULTS: IGF-1 was up-regulated by finasteride treatment in 4 of 9 patients. Among the patients with increased IGF-1 expression, 3 of them showed moderate clinical improvement after 12 months of treatment and another patient remained unchanged. In contrast, 3 patients with decreased IGF-1 expression in the balding scalp showed clinical worsening after 12 months. The other 2 patients without noticeable change in IGF-1 expression showed either slight improvement or no change in their hair condition. CONCLUSION: In a small uncontrolled study of 9 patients with AGA, an increased expression of IGF-1 messenger RNA levels in the DP was associated with patient response to finasteride. PMID- 12894071 TI - Evaluation of the efficacy of antihistamines using human monocyte-derived dendritic cells stimulated with histamine. AB - BACKGROUND: It has been reported that histamine induces CD86 expression and chemokine production in human immature monocyte-derived dendritic cells (MoDCs), which can be blocked by both H(1)- and H(2)-receptor antagonists. OBJECTIVE: We sought to examine whether the efficacy of H(1)-type antihistamines can be assessed by using MoDCs. METHODS: We examined the suppressive effects of 1 H(2) type antihistamine (cimetidine) and 5 different H(1)-type antihistamines (cetirizine, diphenhydramine, ketotifen, olopatadine, and emedastine) on the induction of CD86 and IL-8 production by MoDCs from 23 healthy individuals stimulated with histamine. We also examined the responses of MoDCs from 13 patients with chronic urticaria to these antihistamines, and compared the in vitro efficacy with the actual clinical response to antihistamines evaluated by patient and physician assessments. RESULTS: All the antihistamines we examined suppressed the increase of CD86(+) cells after histamine stimulation in a dose dependent fashion, and all H(1)-type antihistamines were more efficacious than cimetidine. IL-8 production stimulated with histamine was also suppressed by cetirizine, ketotifen, and olopatadine. Unexpectedly, the suppressive effect of these antihistamines on the CD86 augmentation was highly variable among different healthy control participants. Interestingly, in 10 of 13 cases of chronic urticaria, this in vitro analysis of antihistamines correlated with the clinical response to antihistamines. CONCLUSION: This study suggests that the evaluation of antihistamines using MoDCs can be a useful method for the screening of effective antihistamines, for the comparison of the efficacy of antihistamines, and for predicting the efficacy of antihistamines on an individual basis. PMID- 12894072 TI - Evaluation of the histologic characteristics of patch test confirmed allergic contact dermatitis. AB - BACKGROUND: Microscopic features of experimentally induced allergic contact dermatitis (ACD) have been reported; however, the histologic characteristics of environmentally induced ACD have not been definitively elucidated. OBJECTIVE: This pilot study was conducted to describe the histologic features most useful in differentiating ACD from other forms of eczematous dermatitis. METHODS: A retrospective chart review of patients who underwent evaluation for contact dermatitis was performed. The results of skin biopsy were reviewed in a blinded manner, and two subgroups were compared: gold standard ACD cases and control cases (eczematous dermatitis with negative results of patch testing). RESULTS: Of 317 cases reviewed, 39 fulfilled the inclusion criteria. Many histologic similarities were noted: both subgroups had substantial acanthosis and lymphocytic infiltration. In addition, present to a moderate degree in both groups were hyperkeratosis, spongiosis, eosinophils, and dermal dendritic fibrohistiocytic (DFH) cells. However, compared with the control cases, eosinophilic spongiosis and multinucleate dermal DFH cells were found to a considerably greater degree in the standard ACD cases. CONCLUSIONS: Eosinophilic spongiosis and multinucleate dermal DFH cells, in the presence of acanthosis, lymphocytic infiltrate, dermal eosinophils, and hyperkeratosis, are particularly suggestive of ACD. While such findings alone are not diagnostic of ACD, the presence of these combined histologic features supports the pursuit of patch testing in cases of chronic eczematous dermatitis. PMID- 12894073 TI - Hospitalization for severe skin disease improves quality of life in the United Kingdom and the United States: a comparative study. AB - BACKGROUND: Financial and managerial constraints have resulted in the rationalization of dermatology inpatient services in the United Kingdom and the United States. Therapeutic regimes may vary locally, regionally, and internationally but the clinical outcome of treatment remains the same. OBJECTIVE: We studied 2 inpatient units: the University of Wales College of Medicine, Cardiff, United Kingdom, and the University of Miami School of Medicine, Miami, Florida, to compare the use and effectiveness of the service provided. METHODS: Data were collected prospectively from inpatients during a 12 month period. The Dermatology Life Quality Index was administered on admission and after discharge. Data were recorded about the diagnosis, duration of admission, and referring dermatologist. RESULTS: In all, 295 patients (Cardiff, UK) and 366 patients (Miami, Fla) participated. The average duration of admission in Miami was 6.7 days compared with 14.2 (P <.0001) in Cardiff. In Miami, the most common reasons necessitating admission were extensive disease (54%), the patient being unwell (18%), photophoresis (14%), outpatient treatment failure (8%), and acute deterioration of disease (4%). In Cardiff, the common reasons were acute deterioration (35%), extensive disease (28%), outpatient treatment failure (22%), and liver biopsy (4%). The most common diagnoses in Cardiff were psoriasis (31%) and eczema (26%). In contrast, the most common diagnoses in Miami, were psoriasis (19%), leg ulcers (17%), and mycosis fungoides (14%). The mean Dermatology Life Quality Index value for all patients decreased after admission in Cardiff (14.9-8.2, P <.0001) and Miami (12.0-8.5, P <.0001). CONCLUSION: Despite the differences in the 2 health care systems, inpatient therapy remains an important and effective therapeutic option in the United States and the United Kingdom. PMID- 12894074 TI - Immunologic escape and angiogenesis in human malignant melanoma. AB - BACKGROUND: Melanoma escape mechanisms include immunosuppressive and angiogenic cytokine production. OBJECTIVE: We sought to determine vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) expression by immunohistochemistry, and soluble circulating plasma levels of VEGF, bFGF, IL-10, and transforming growth factor-beta2 in patients with different stages of melanoma. METHODS: Biopsy specimens from 42 patients with primary melanoma and 9 with cutaneous metastases were studied by immunohistochemistry. In another 46 patients with melanoma (8 stage I and II; 18, III; and 20, IV) and in 10 healthy control participants, bFGF, VEGF, IL-10, and transforming growth factor-beta2 circulating levels were analyzed. RESULTS: bFGF was positive in 85% and VEGF in 47.5% of 42 primary melanomas. Of 10 patients with primary melanoma (Breslow depth 1.5-3 mm) 6 were VEGF positive and had metastases develop, whereas 4 were VEGF negative and had no metastases at 5 years of follow up. VEGF, bFGF, and IL 10 plasma levels in patients with stages III and IV melanoma were higher than the control group (P <.05 and P <.01, respectively). An inverse relationship was found between VEGF and IL-10. Specifically, in 7 patients with IL-10 levels higher than 10 pg/mL, VEGF levels were less than 49 pg/mL (P <.05); in 9 patients with VEGF levels higher than 100 pg/mL, IL-10 levels were less than 6.7 pg/mL (P <.01). CONCLUSION: VEGF expression in 1.5- to 3.0-mm Breslow depth melanomas may be considered as an unfavorable prognostic factor. Immunosuppressive (IL-10, transforming growth factor-beta2) and proangiogenic (bFGF, VEGF) cytokines are increased in metastatic melanoma. Inverse plasma levels between IL-10 and VEGF in patients with metastatic melanoma are shown in vivo for the first time, the significance of which must be further investigated. PMID- 12894075 TI - Hypopigmented mycosis fungoides in Caucasian patients: a clinicopathologic study of 7 cases. AB - BACKGROUND: Hypopigmented mycosis fungoides (MF) is a rare variant of cutaneous T cell lymphoma. It is more frequent in dark-skinned or Asian patients, particularly children. Only 9 cases in Caucasian patients have been reported in the literature so far. OBSERVATION: We describe 7 Caucasian patients (2 children and 5 adults) with hypopigmented MF. Histologic examination confirmed the diagnosis in all cases. The phenotype of neoplastic lymphocytes was T helper in 4 cases and T suppressor in 3 (2 of them children). Monoclonality of the T lymphocytes could be detected in hypopigmented lesions in all 7 cases with the use of a polymerase chain reaction technique. In 4 patients, polymerase chain reaction analysis of T-cell receptor-gene rearrangement after laser-based microdissection of the specimen revealed that the monoclonal population of T lymphocytes was confined mainly to the epidermis. CONCLUSION: Hypopigmented lesions of MF can be observed in Caucasian patients. Children affected by MF often present with this rare clinical variant of the disease. Persistent or unusual hypopigmented lesions should be subjected to biopsy to avoid delay in the diagnosis of MF, especially in children. PMID- 12894076 TI - Patients with palmoplantar psoriasis have more physical disability and discomfort than patients with other forms of psoriasis: implications for clinical practice. AB - BACKGROUND: Psoriasis is a chronic, unpredictable, and incurable disease that has a negative impact on patients' quality of life. Palm and sole psoriasis can add to this negative impact as it directly affects activities of daily living. OBJECTIVE: We sought to estimate the prevalence of palmoplantar psoriasis in a patient population and to explore associations with patient outcomes. METHODS: In all, 317 individuals with psoriasis completed a comprehensive assessment battery. Patients with palmoplantar psoriasis (n = 124, 39%) were compared with patients without palmoplantar involvement with respect to functional disability, psychiatric symptoms, physical and social discomfort, self-reported psoriasis severity, and health-related quality of life. RESULTS: Patients with palmoplantar involvement reported significantly greater physical disability and physical discomfort than patients without palmoplantar involvement (both P <.01). There were no differences between the 2 groups with respect to psychosocial outcomes. CONCLUSION: Patients with palmoplantar psoriasis are affected to a greater degree by the physical aspects of the disease than patients without palmoplantar involvement. PMID- 12894077 TI - Oral cyclophosphamide for treatment of pemphigus vulgaris and foliaceus. AB - BACKGROUND: Cyclophosphamide is an alkylating adjuvant used in refractory cases of pemphigus. OBJECTIVE: We sought to evaluate the effectiveness and safety of oral cyclophosphamide in the treatment of patients with pemphigus vulgaris (PV) and pemphigus foliaceus (PF) with refractory disease. PATIENTS: We studied 23 patients with pemphigus (20 with PV; 3 with PF) who failed to achieve clinical remissions with the use of prednisone and antimetabolites. RESULTS: Complete remission was achieved in 17 patients with PV and 2 with PF. A total of 3 patients with PV failed therapy. A partial remission was achieved in 1 patient with PF. The treatment was administered for a median duration of 17 months with a follow-up period of 27 months. The median time to complete remission was 8.5 months. A total of 9 patients who were severely affected received concomitant plasma exchange. Adverse reactions included 5 cases of hematuria, 6 nonlife threatening infections, and the development of transitional cell carcinoma of the bladder 15 years after discontinuation of cyclophosphamide in 1 patient. No death was associated with cyclophosphamide treatment. CONCLUSION: Oral cyclophosphamide is an effective adjuvant in the treatment of severe and refractory PV and PF, but requires close monitoring. PMID- 12894078 TI - Systemic therapy with cyclophosphamide and anti-CD20 antibody (rituximab) in relapsed primary cutaneous B-cell lymphoma: a report of 7 cases. AB - BACKGROUND: Rituximab, a chimeric antibody directed against CD20, has a high therapeutic value in refractory/relapsed low-grade or follicular B-cell non Hodgkin's lymphomas as a monotherapy or in combination with polychemotherapy. OBJECTIVES: We sought to evaluate the clinical activity and toxicity of a schedule foreseeing the use of intravenous rituximab preceded by single-dose cyclophosphamide in the treatment of patients with primary cutaneous B-cell lymphoma who progressed and relapsed after chemotherapy. METHODS: A total of 7 patients were treated; 4 had both cutaneous lesions and nodal or visceral involvement. All the patients had been previously treated with at least 1 standard chemotherapy regimen, and 4 with 2 or more, with a median response duration of 8 months. Immunohistochemistry on frozen sections was performed with monoclonal antibodies directed against CD20, CD55, and CD59 before rituximab treatment. RESULTS: The overall objective response rate was 85.7%, with a complete response in 5 patients; treatment was well tolerated in all cases. After a median follow-up of 13 months, 2 patients showed a cutaneous relapse. The response durations of the remaining patients who were disease-free are now 5, 7, 17, and 18 months. No relationship was found between CD55 and CD59 expression and the clinical outcome. CONCLUSION: Although a longer follow-up period is needed to confirm these data, our results are encouraging, particularly in terms of disease free survival. PMID- 12894079 TI - Surgical pearl: hands-free tape retraction. PMID- 12894080 TI - Topical nitrogen mustard in the treatment of alopecia areata: a bilateral comparison study. AB - Topical nitrogen mustard is an alkylating agent. Its efficacy in treating alopecia areata was reported in an uncontrolled study. We present a preliminary, half-head, controlled 16-week study showing that topical nitrogen mustard was of benefit in 1 of 6 patients treated with 50% to 100% scalp involvement. Another 4 patients did not complete the trial. PMID- 12894081 TI - Fatal interstitial pulmonary fibrosis in anti-Jo-1-negative amyopathic dermatomyositis. AB - Amyopathic dermatomyositis (DM) describes a subpopulation with the cutaneous eruption of DM, but without muscle involvement. Interstitial pulmonary fibrosis is a recognized complication of DM, often correlated with antisynthetase enzymes, such as anti-Jo-1. We describe a case of fatal IPF in a patient with anti-Jo-1 antibody-negative amyopathic DM. PMID- 12894082 TI - Blau syndrome: a new kindred. AB - Blau syndrome is a rare condition typically defined by granulomatous arthritis, skin eruption, and uveitis occurring in the absence of lung or other visceral involvement. Other characteristic physical findings include synovial cysts and camptodactyly. We describe a new kindred demonstrating autosomal dominant inheritance and anticipation. PMID- 12894083 TI - A case of chronic herpes gestationis: persistent disease or conversion to bullous pemphigoid? AB - We report the case of a 38-year-old woman with herpes gestationis (HG) persistent for 26 months postpartum with typical erythematous-edematous grouped lesions associated with vesicles, blisters, and scaled crusts on most parts of her body. Despite high doses of oral prednisolone, azathioprine, and dapsone, and a trial of 5 plasmaphereses, the disease has persisted to date. Histopathologic examination of lesional skin showed subepidermal blisters, focal basal cell necrosis, and a dermal inflammatory infiltrate including many eosinophils. Direct immunofluorescence showed linear C3 staining in the basement membrane zone and the complement fixation test demonstrated circulating antibasement membrane zone antibodies at a low titer (1:80). HLA typing demonstrated an A2, A24, B35, B52, DR4,5, DR13,15, DRW52,53 phenotype. We present this case as chronic persistent HG and discuss the differential diagnosis between chronic persistent HG and HG evolving to bullous pemphigoid, together with a careful examination of similar cases reported in literature. PMID- 12894084 TI - Cutaneous nodular amyloidosis masquerading as a foot callus. AB - We report a unique case of a 69-year-old man who presented with a 1-year history of pink nodules and plaques limited to the anterior plantar surface of the right foot and a 30-year history of a callus on the right heel. Histologic examination findings of both areas showed deposits of amorphous, eosinophilic material and an infiltrate of plasma cells in the dermis. Congo red-stained deposits exhibited apple-green birefringence with polarized light. Results of an extensive clinical and laboratory evaluation showed no evidence of systemic amyloidosis. The diagnosis of nodular primary localized cutaneous amyloidosis (PLCA) was made. Nodular PLCA isolated to pedal or strictly plantar surfaces is an unusual presentation of PLCA. The origin and the clinical and histopathologic features of nodular PLCA are reviewed. PMID- 12894085 TI - Subungual blue nevus. AB - Junctional or compound melanocytic nevus and melanoma may involve the nail apparatus and are usually discussed in the differential diagnosis of nail pigmentation. Blue nevus has been very rarely reported in this location. We report the second clinicopathologic study of a subungual blue nevus. PMID- 12894086 TI - Oral pemphigus vulgaris associated with HIV infection. AB - Vesiculobullous disease in association with HIV infection is extremely rare. This report details the presentation and management of pemphigus vulgaris in an individual with HIV infection. The clinical characteristics and response to therapy appear not to be modified by coexistent HIV infection. However, the management of pemphigus vulgaris relies on immunosuppressive therapy potentially resulting in HIV disease progression. Cyclosporine has in vitro antiretroviral activity and, currently, should be considered as a possible treatment for individuals with pemphigus vulgaris and HIV infection. PMID- 12894087 TI - Fatal sodium valproate-induced hypersensitivity syndrome with lichenoid dermatitis and fulminant hepatitis. AB - Anticonvulsant hypersensitivity syndrome caused by sodium valproate (Depakene) has rarely been reported. Here we describe a patient who presented with nonspecific polymorphous eruptions, fulminant hepatitis, and jaundice. Lichenoid dermatitis was found in the skin biopsy specimen. Toxic hepatitis and microvesicular steatosis were found in the liver biopsy specimen, consistent with valproate hepatotoxicity. PMID- 12894088 TI - Primary jejunal malignant melanoma first noticed because of the presence of parotid lymph node metastasis. AB - The case of a 71-year-old Japanese man with primary jejunal melanoma with an unusual clinical course is reported. Several cases of primary malignant melanoma of the small intestine have been reported, but in most cases no clear evidence has shown whether the lesions were primary or metastatic. We discuss why the tumor in our case was considered primary, and we review similar cases of primary melanoma of the small intestine. PMID- 12894089 TI - Superficial morphea. AB - The superficial variant of morphea is a recently described entity that differs in its clinical and histologic presentation from classic morphea. The condition has been reported only in females to date and is characterized by minimal to no induration and hypopigmented or hyperpigmented patches without associated symptoms, systemic involvement, contractures, or atrophy. We present the unique clinical and histopathologic findings in a 77-year-old woman. PMID- 12894090 TI - Necrolytic migratory erythema as the only presenting sign of a glucagonoma. AB - We describe a 39-year-old man with a 3-year history of a recalcitrant psoriasiform eruption that was accentuated in the intertriginous areas. Hsitopathology was consistent with psoriasis. A glucagon level was 744 pg/mL with the upper limit of normal being 130 pg/mL. Computed tomographic scan of the abdomen revealed a 5-cm mass in the tail of the pancreas. The tumor was removed and found to be a glucagonoma (pancreatic islet tumor). The clinical eruption resolved promptly with surgical excision. Neither the clinical eruption nor the tumor has recurred for 6 months. The course of disease confirms the diagnosis of necrolytic migratory erythema associated with a glucagonoma. PMID- 12894091 TI - Treatment of pustular psoriasis with granulocyte and monocyte adsorption apheresis. AB - We studied the efficacy of granulocyte and monocyte adsorption apheresis in 2 patients with pustular psoriasis, one localized, the other generalized. Treatment with granulocyte and monocyte adsorption apheresis resulted in remarkable clearing of the skin lesions, suggesting that this therapy is a valuable tool for treating patients with intractable skin diseases attributable to activated granulocytes. We present detailed descriptions of these patients and this novel therapy. PMID- 12894092 TI - Nevoid basal cell carcinoma syndrome in a person with dark skin. AB - The expression of basal cell carcinoma tumors of the skin is blunted in individuals with dark skin and nevoid basal cell carcinoma syndrome. The occurrence of multiple basal cell carcinomas in these patients is a relatively rare finding. We describe a 25-year-old man of partial African-American descent with constitutive Fitzpatrick type IV pigmented skin and the clinical stigmata of nevoid basal cell carcinoma syndrome including histopathologic evidence of 11 basal cell carcinomas. PMID- 12894093 TI - Strawberry gingiva: a distinctive sign in Wegener's granulomatosis. AB - A 37-year-old man presented with gingival hyperplasia accompanied by an ulcer on the eyelid, nasal obstruction, ear discharge, and discharging nodules on the cheek and back. An evaluation for infectious diseases and leukemia was negative. During his hospital stay, he had fever and migratory joint pains develop, suggesting a diagnosis of Wegener's granulomatosis, which was confirmed by a strongly positive cytoplasmic antineutrophil cytoplasmic antibody test. He responded promptly to treatment with oral prednisolone and cyclophosphamide. In retrospect, the diagnosis could have been suspected earlier because of the distinctive red, friable, and granular appearance of the gingiva. PMID- 12894094 TI - Syringoma of the moustache area. AB - Syringomas are benign tumors of the eccrine ducts that appear as small skin colored or yellow papules. They usually occur in the periorbital area, but have also been found on the scalp, forehead, cheeks, axillae, abdomen, extremities, genitalia, and buttocks. We describe a patient with an unusual presentation of syringomas on the moustache area. PMID- 12894095 TI - Hydroxyurea-induced melanonychia concomitant with a dermatomyositis-like eruption. AB - We describe the cases of 7 patients with hydroxyurea-induced melanonychia. In two of the patients, a dermatomyositis-like eruption was also present. PMID- 12894096 TI - Metastatic hepatocellular carcinoma of gingiva mimicking pyogenic granuloma. AB - We report a case of metastatic hepatocellular carcinoma of gingiva mimicking pyogenic granuloma. To our knowledge, no such reports have been published in the literature of dermatology. Gingival metastatic lesions are of interest both because they are easily mistaken for several benign lesions and because they may be the first sign of an undiscovered malignancy. PMID- 12894097 TI - Penicillium marneffei infection diagnosed by polymerase chain reaction from the skin specimen. AB - We describe a case of Penicillium marneffei infection in a patient with AIDS who had skin eruptions disseminated over the entire surface of his skin. We identified P marneffei from the skin biopsy specimen by polymerase chain reaction using a set of primers specific for this pathogen. PMID- 12894098 TI - Bullous scabies. AB - Bullous scabies is a rare presentation usually occurring in patients who are elderly. Clinical, histologic, and immunofluorescent findings mimic bullous pemphigoid. We report a case of bullous scabies in a 4-year-old boy and review the literature. PMID- 12894099 TI - New psoriasis treatments based upon a deeper understanding of the pathogenesis of psoriasis vulgaris and psoriatic arthritis: a personal appraisal of their use in practice. PMID- 12894100 TI - Newer antifungal agents. PMID- 12894101 TI - Etanercept and squamous cell carcinoma. PMID- 12894102 TI - Surgical pearl: accurate documentation of facial lesions using only one landmark. PMID- 12894103 TI - Ratio of irritant contact dermatitis to allergic contact dermatitis in occupational skin disease. PMID- 12894104 TI - Familial Laugier-Hunziker syndrome. AB - We report the unusual occurrence of 3 cases of Laugier-Hunziker syndrome in the same family. PMID- 12894105 TI - Perforating pilomatricoma in a process of total elimination. AB - Perforating pilomatricoma is a rare clinical variant of pilomatricoma. We report a case of pilomatricoma with ulceration indicating total elimination. The lesion consisted mainly of laminating shadow cells surrounded by invaginating epidermis. We believe that this case can be considered within the category of perforating pilomatricoma. PMID- 12894106 TI - Infantile myofibromatosis: support for autosomal dominant inheritance. AB - We describe a family in whom infantile myofibromatosis affected 3 generations. The disease expression in this family suggests an autosomal dominant inheritance pattern with variable penetrance. PMID- 12894107 TI - Infectious causes of pityriasis lichenoides: a case of fulminant infectious mononucleosis. AB - Pityriasis lichenoides is a rare cutaneous eruption of unknown cause that spans a spectrum of clinical severity. Infectious agents have long been suspected as etiologic factors. The present case is the first to demonstrate a known EBV mediated process evolving and resolving in concert with pityriasis lichenoides. Epstein-Barr virus, Toxoplasma gondii, and HIV are the most frequently reported infectious triggers of pityriasis lichenoides. Pityriasis lichenoides may arise secondary to EBV-mediated acute infectious mononucleosis. PMID- 12894108 TI - Histopathologic and scanning electron microscope examination of the nail and hair in chronic mucocutaneous candidiasis. AB - The case of a 4-year-old boy with chronic mucocutaneous candidiasis is reported. Nail sections stained with periodic acid-Schiff and Grocott-Gomori methenamine silver showed pseudohyphae running haphazardly within the nail. Scanning electron microscopy showed hyphae penetrating into the nail plate and the hair shaft. PMID- 12894109 TI - Cutaneous Strongyloides stercoralis infection: an unusual presentation. AB - Strongyloides stercoralis is a widespread, soil-transmitted, intestinal nematode common in tropical and subtropical countries. The parasite is unique in its capability to carry out its entire life cycle inside the human body. Human beings contract strongyloidiasis by penetration of filariform larvae into the skin or mucous membrane after contact with contaminated soil. The larvae travel by the venous systems to the lungs, then ascend the bronchi to the trachea, where the larvae are coughed up by the human host, subsequently swallowed, and attain their habitat in the small intestine. Chronic strongyloidiasis acquired in endemic areas may last decades and gives rise to various dermatologic lesions, the most characteristic of which is larva currens, a serpiginous, creeping urticarial eruption. In disseminated strongyloidiasis, the characteristic skin lesions are widespread petechiae and purpura. We present a case of disseminated strongyloidiasis with an unusual manifestation mimicking a drug rash and review the dermatologic manifestations of strongyloidiasis infestation. PMID- 12894110 TI - Acute bullous purpura associated with hyperhomocysteinemia and antiphospholipid antibodies. AB - We describe a female patient with an acute purpuric and bullous eruption mainly affecting the lower aspect of the legs. Skin biopsy specimens demonstrated microvascular occlusions with fibrin thrombi but no dermal inflammation. Intermediate hyperhomocysteinemia and transient antiphospholipid antibodies were evidenced as factors of thrombophilia. The responsibility of the latter in the onset of the cutaneous lesions is discussed. PMID- 12894111 TI - Segmental tuberous sclerosis presenting as unilateral facial angiofibromas. AB - Tuberous sclerosis (TSC) is an autosomal dominant inherited disorder with cutaneous lesions of ash leaf hypopigmented macules, shagreen patches, periungual fibromas, facial angiofibromas, forehead fibrous plaques, confetti hypopigmentation, and poliosis. Multiple facial angiofibromas are a pathognomonic feature of TSC. Unilateral facial angiofibromas, however, represent a rare variant of TSC, as only 6 cases are reported in the literature. We describe a case of a 52-year-old man who presented with unilateral facial angiofibromas and poliosis without other features of TSC. PMID- 12894112 TI - Phakomatosis pigmentovascularis IIb with hypoplasia of the inferior vena cava and the right iliac and femoral veins causing recalcitrant stasis leg ulcers. AB - We present a patient with phakomatosis pigmentovascularis (PPV) type IIb accompanied with venous hypoplasia, from the inferior vena cava to the superficial femoral vein. Although it is not obvious whether this vascular anomaly, which is probably congenital, is coincidental or not, it is possible that the 2 diseases have some relationship each other, because PPV is thought to result from abnormal vasomotor activity during the embryonic period. In the diagnosis of type II PPV, careful examination and several studies are required to determine systemic involvement that may include large vessel changes. PMID- 12894113 TI - Branched filaments no fungus, ovoid bodies no bacteria: Two unusual cases of mycetoma. AB - We describe a 58-year-old man presenting with necrotizing panniculitis of the lower right leg and a 64-year-old woman with a clinically similar lesion combined with pustular eruptions and subsequent ulceration on the forehead. In the first patient, Giemsa staining showed small ovoid bodies and Grocott staining revealed hyphae. Histology from the process on the forehead showed branched filaments in the periodic acid-Schiff (PAS) staining. In the first case, Madurella mycetomatis, a fungus, was the pathogenic agent, whereas in the other case white colonies of filamentous organisms resembling fungi could be cultivated that turned out to be the bacterium Nocardia brasiliensis. Since the initial clinical appearance of these two forms of mycetoma were almost identical and histopathologic findings were inconclusive, only sophisticated microbiologic work up of material from lesional skin led to the correct diagnosis. In times of global tourism, these unusual cases impressively document the necessity to become more familiar with mycetoma to make accurate therapeutic decisions with effective results, possibly saving a limb. PMID- 12894114 TI - Secondary syphilis presenting as pseudolymphoma of the skin. AB - Secondary syphilis most commonly presents with a papulosquamous eruption that involves the palms, soles, and mucous membranes. The papulonodular variant has only been described 11 times in the literature. We describe a case of papulonodular secondary syphilis presenting as an atypical lymphoid hyperplasia suggestive of cutaneous lymphoma. PMID- 12894115 TI - Tinea capitis in adult women masquerading as bacterial pyoderma. AB - Tinea capitis is generally thought to be a common disease in children but not in adults. Adults with tinea capitis generally present with scale and alopecia. We report 3 adults with inflammatory tinea capitis caused by Trichophyton tonsurans that resembled a bacterial infection. Of these patients, 2 were initially given a diagnosis of bacterial pyoderma. All patients were successfully treated with oral antifungal agents. One patient had significant eosinophilia that resolved with treatment. We conclude that tinea capitis should remain in the differential diagnosis of adults with alopecia and pyoderma-like presentations. A biopsy specimen was helpful in making the diagnosis in 2 of the 3 patients, but fungal culture confirmed the diagnosis in all cases. PMID- 12894116 TI - Cutaneous sarcoidlike lesions in B-cell chronic lymphocytic leukemia. AB - The development of systemic sarcoidosis in patients with malignancies is a well known phenomenon. However, cutaneous sarcoidlike lesions are a rare finding. We report 2 patients with B-cell chronic lymphocytic leukemia and hypogammaglobulinemia who presented isolated cutaneous sarcoidlike lesions 9 years after the diagnosis of the hematologic malignancy in one case and after the second course of chemotherapy regimen in the other case. The role of cytokines released from malignant cells and of the hypogammaglobulinemia facilitating an unidentified infectious agent is questionable. PMID- 12894117 TI - Crohn's disease presenting as prepuce and scrotal edema. AB - Metastatic cutaneous Crohn's disease, in which noncaseating granuloma infiltration of the skin occurs at sites separated from the gastrointestinal tract by normal tissue, is the least common dermatologic manifestation of Crohn's disease. We report a case of a 34-year-old man with metastatic Crohn's disease presenting as prepuce and scrotal edema with typical histopathologic features. We think that any unusual cutaneous lesion in patients with Crohn's disease should be biopsied. PMID- 12894118 TI - Granuloma annulare masquerading as molluscum contagiosum-like eruption in an HIV positive African woman. AB - A 33-year-old African woman who was HIV positive and being treated with zidovudine and zalcitabine presented with a 4-week history of a generalized pruritic rash superficially resembling molluscum contagiosum. The appearance of the lesions appeared to coincide with a dramatic decrease in her peripheral CD8+, and to a lesser extent, CD4+ T-lymphocyte count. Hematologic investigations revealed anemia with eosinophilia and she had a strongly positive Strongyloides antibody test. The eruption persisted despite appropriate antihelminthic treatment and temporary withdrawal of antiretroviral therapy. Histologic examination showed ill-defined nodules of necrobiotic collagen with surrounding palisading mononuclear cell infiltrate consistent with granuloma annulare. The rash disappeared spontaneously over several weeks. PMID- 12894119 TI - Neonatal pemphigus foliaceus. AB - The term "pemphigus" refers to a group of diseases that are characterized by the presence of cutaneous or mucosal blisters and erosions, and antiepidermal autoantibodies. There are several case reports of neonatal pemphigus vulgaris in the literature. Although pemphigus foliaceus antibodies have been shown to cross the placenta, to our knowledge, this is only the second reported case of neonatal pemphigus foliaceus. The proposed mechanism of disease transfer is the passive transfer of maternal IgG antibodies across the placenta. PMID- 12894120 TI - Cutaneous findings in a 51-year-old man with phenylketonuria. AB - A man with phenylketonuria who was reported to have lichen sclerosus et atrophicus at age 16 years presented to our institution 35 years later with large confluent areas of atrophy. Although sclerodermatous changes have been described in children with phenylketonuria, this case offers a longitudinal view of the progression of skin lesions in a middle-aged man with phenylketonuria. PMID- 12894121 TI - Spontaneous dermal abscesses and ulcers as a result of Serratia marcescens. AB - Serratia sp have only rarely been reported as isolates from leg ulcers. We describe the case of a middle-aged man with a medical history significant for alcohol-induced cirrhosis who presented with rapidly progressive skin ulcers initially starting as purple nodules. These skin ulcers and underlying dermal abscesses were found to be a result of S marcescens, with the presumed portal of entry being a toe-web infection. PMID- 12894122 TI - Angiolymphoid hyperplasia with eosinophilia responsive to pulsed dye laser. AB - Angiolymphoid hyperplasia with eosinophilia is an uncommon, benign, but potentially disfiguring vascular proliferation. Angiolymphoid hyperplasia with eosinophilia commonly affects women in the third decade. Although a benign process, significant morbidity can occur including bleeding, pain, and disfigurement. Several therapies have been attempted including curettage, cryotherapy, retinoids, excision, and various lasers. We report on a case of angiolymphoid hyperplasia with eosinophilia responding to pulsed dye laser therapy (585 nm) (SPTL-1a, Candela Corp, Wayland, Mass). PMID- 12894123 TI - Cutaneous ulceration caused by methotrexate. AB - Methotrexate has been associated with the development of skin ulceration in small numbers of patients with psoriasis. We describe a 58-year-old man with a 15-year history of plaque-type psoriasis. He had been treated with methotrexate therapy (15 mg orally per week for 32 weeks) and had ulcers develop within psoriatic plaques on both elbows, on the mucosal aspect of the lower lip, and in an appendectomy scar on the right lower quadrant of the abdomen. Ulcers healed upon discontinuation of methotrexate, but the scar eroded again upon reintroduction of methotrexate. PMID- 12894124 TI - Psoriasis treatment 2003 at the new millennium: position paper on behalf of the authors. PMID- 12894125 TI - The immunology of psoriasis and biologic immunotherapy. AB - In this review, we will discuss the immunological basis for psoriasis with special emphasis on the role of effector T cells. With the understanding of this immunologic process, we will present a model for the development of targeted immune response modifiers, termed biologic immunotherapies, and their potential role for the benefit of patients with psoriasis. PMID- 12894126 TI - Genetics of psoriasis: the potential impact on new therapies. AB - There is considerable epidemiologic evidence that genes play a key role in the pathogenesis of psoriasis. It is also clear that multiple genes are involved and that the disease is genetically heterogeneous. Important interactions with the environment are also implicated in its development. A number of genetic loci have been identified by genome wide linkage scans and two loci have been replicated: PSORS1 on chromosome 6, within the major histocompatibility complex, and PSORS2 on chromosome 17q. Understanding the genetic basis of psoriasis will represent a major advance in our understanding of the disease and will reveal novel disease specific biologic pathways. This information will be used to develop more specific diagnostic and prognostic tools and also lead to the development of individualized treatment plans. Benefits of the latter include more effective and safer treatments and potentially major pharmaco-economic gain. PMID- 12894127 TI - Quality of life issues in psoriasis. AB - Psoriasis is associated with significant psychosocial morbidity and a decrease in health-related quality of life. It is important to view psoriasis as a serious disease and resist the tendency to underestimate its impact on overall patient well-being. The disability experienced by psoriasis sufferers is comparable to that of patients with other chronic illnesses such as heart disease, diabetes, cancer, and depression. Aggressive intervention is warranted in order to improve patient quality of life and decrease the potential for psychosocial sequelae. Health-related quality of life measures are becoming a necessary adjunct to traditional clinical assessments in the evaluation and treatment of psoriasis patients by the individual clinician. They also provide valuable information to government agencies and third party payers in the determination of resource allocation and reimbursement. PMID- 12894128 TI - The design of clinical trials in psoriasis: lessons for clinical practice. AB - Understanding the process by which drugs are approved gives clinicians a better understanding of the medications they prescribe and how these medications are best used to care for patients. The development of new drugs in the United States proceeds through a series of studies culminating in well-designed, large-scale human trials. These studies result from a coordinated collaboration between industry and the Food and Drug Administration (FDA). The results of these studies determine whether drugs are approved for use or not. They also determine the labeling and marketing of products and guide (but do not fully determine) physicians' use of the drug. This article follows a hypothetical drug for use in psoriasis through the approval process, in order to illustrate the process and its implications for clinical practice. PMID- 12894129 TI - Current systemic therapies for psoriasis: where are we now? AB - Many systemic agents are used in the treatment of psoriasis. They provide good control of psoriasis in the majority of patients and have improved their life quality indices. Frequently, combination therapy is utilized to synergize the efficacy of these medications. Many dermatologists are hesitant in prescribing systemic agents because of their adverse effects. However, such potential toxicities arising from these medications can largely be avoided if proper patient selection and close monitoring are performed. PMID- 12894130 TI - Phototherapy treatment of psoriasis today. AB - The use of the various forms of phototherapy remains an essential treatment option for psoriasis vulgaris. Expertise concerning the mechanisms involved with the actions of therapeutic ultraviolet light and the proper delivery of office based treatments resides within the specialty of dermatology. New therapies for the treatment of moderate to severe psoriasis will soon become available which have specific actions on the cutaneous immune system. A better understanding of the known mechanisms of action for ultraviolet light therapy makes it appropriate to include this area of treatment with new biologic agents. Photochemotherapy and various forms for delivery of narrow band ultraviolet B can be used as treatments, either as monotherapy or in combination with other agents, to effectively treat moderate and severe psoriasis. PMID- 12894131 TI - Development and use of alefacept to treat psoriasis. AB - Activated memory T cells, expressing CD2, are key components in the pathogenesis of psoriasis. Alefacept binds to CD2, blocks co-stimulatory signaling, and selectively induces apoptosis of pathogenic T cells. Our objective is to present safety and efficacy results which lead to the new drug application (NDA) of alefacept for the treatment of psoriasis. We reviewed the key phase II and III trials in over 1300 patients and found that during treatment and follow-up of patients receiving 12 weekly intramuscular or intravenous injections of alefacept, about 1/3 will achieve a reduction in psoriasis area and severity index (PASI) of > or =75% and nearly 2/3 a reduction in PASI of > or =50%. Patients who achieved a > or =75% reduction from baseline PASI during or after a single course maintained a > or =50% reduction in PASI for a median duration of >7 months. Among patients who received 2 courses of alefacept, 40% and 71% of patients achieved a > or =75% and > or =50% reduction in PASI, respectively and duration of effect was prolonged. Adverse events in the placebo and active treatment arms did not differ. We conclude that alefacept significantly improves psoriasis and produces durable clinical improvement with a very favorable safety profile. PMID- 12894132 TI - Efalizumab: an overview. AB - Efalizumab (anti-CD11a), a targeted T cell modulator, is a humanized monoclonal antibody being studied for plaque psoriasis. Phase I and II studies demonstrated that efalizumab treatment results in histologic improvement and clinical benefit in patients with moderate to severe disease. Phase III clinical trials confirmed the clinical activity and safety of weekly efalizumab. Efalizumab is associated with a rapid onset of clinical benefit, with significant improvements observed as early as 4 weeks. Extending treatment from 12 to 24 weeks both maintains and improves the initial responses achieved at 12 weeks. Both 12 and 24 weeks of efalizumab therapy were generally well tolerated. Ongoing studies are being conducted to further define the optimal dosing and administration schedules for efalizumab in patients with moderate to severe plaque psoriasis. PMID- 12894133 TI - Etanercept: An overview. AB - Etanercept, a competitive inhibitor of TNF-alpha, is currently FDA approved for psoriatic arthritis and rheumatoid arthritis. The molecular structure of etanercept, its mechanism of action, and results of clinical trials involving patients with psoriasis will be reviewed. PMID- 12894134 TI - Infliximab for psoriasis. AB - This review summarizes the use of inflximab in psoriasis and other immune mediated inflammatory disorders (IMIDs). The magnitude and speed of the response to infliximab monotherapy of moderate to severe psoriasis vulgaris is substantial, being similar to those achieved with cyclosporin. In contrast with cyclosporin, clinical improvement after the initial 3 intravenous influsions of infliximab is maintained for as long as 6 months in approximately half the patients with the absence of any additional treatment. Additionally, infliximab monotherapy normalizes keratinocyte proliferation and differentiation and markedly decreases epidermal inflammation. These results provide a convincing argument for the role of TNF-alpha in the pathogenesis of psoriasis and for the clinical development of infliximab for the treatment of psoriasis. PMID- 12894135 TI - Combining the new biologic agents with our current psoriasis armamentarium. AB - Combination therapy, rotational therapy, and sequential therapy have been used for psoriasis in attempts to achieve greater efficacy and greater safety. The purpose of this manuscript is to review potential advantages and disadvantages of new biologic agents as we look forward to their use in combination regimens with other systemic, topical, and light therapies. Data on the efficacy and toxicity of existing systemic therapies and new biologic agents is reviewed with an emphasis on potential additive or synergistic benefits or toxicities. The mechanism of action of biologic agents differs from systemic agents currently in use, suggesting that there may be additive effects in treating psoriasis. The absence of hepatotoxicity and nephrotoxicity are important advantages when considering combination therapy with biologic agents. The advantages of the use of biologic therapies in combination or rotation with other systemic agents will have to be demonstrated in clinical trials. Mechanisms of action of the biologic therapies suggest that there is potential for additive benefit when used in combination regimens. PMID- 12894136 TI - Evaluation and management of psoriatic arthritis: the role of biologic therapy. AB - Clinicians often view psoriatic arthritis (PsA) as a rather minor arthritic disorder because many are unaware of the substantial damage, disability, and reduced quality of life that patients with this disease can suffer. Compared with better-studied arthritic conditions, such as rheumatoid arthritis (RA) with well known consequences of disease progression, PsA does not elicit the same urgency to treat early and aggressively. This is largely owing to the lack of predictive epidemiologic data regarding disease progression in PsA. However, numerous studies indicate that PsA and RA are comparable in terms of overall severity of joint involvement and disability over equivalent disease duration. Many of the drugs traditionally used for PsA therapy are also used to treat RA, including nonsteroidal anti-inflammatory drugs (NSAIDs), corticosteroids, methotrexate (MTX), sulfasalazine, cyclosporine, etretinate, auranofin, intramuscular gold, and azathioprine. All of these drugs have significant risk of toxicity over long term use, and all provide variable efficacy. This makes it difficult for clinicians to make sound risk-benefit assessments regarding treatment or nontreatment of PsA, because the risks of disease progression cannot be weighed against the risks of therapy. The newer biologic antirheumatic drugs appear to combine greater efficacy of treatment with significantly less toxicity by targeting specific mediators involved in the pathogenesis of PsA. PMID- 12894137 TI - Investigational therapies for psoriasis. AB - As the pathogenesis of psoriasis is better understood, specific targeted therapies are being developed. In addition to alefacept, etanercept, efaluzimab, and infliximab, discussed in separate articles in this issue, numerous other investigational therapies are currently in clinical trials, some of which will likely be approved in the future. We review the most promising of these therapies and their mechanisms of action. PMID- 12894138 TI - Integrating biologic therapies into a dermatology practice: practical and economic considerations. AB - This article discusses the issues involved in the integration of biologic therapies for psoriasis into a dermatology practice. The requirements for staff, space, and other office adaptations are reviewed for infliximab, efalizumab, etanercept, and alefacept in their current injectable forms. Dermatologists will likely elect to offer some or all of these therapies depending on the adjustments necessary within their current practice, patient satisfaction, and the economic possibilities. This article provides information needed to guide dermatology practices in practical decisions regarding the use of biologic therapies. PMID- 12894139 TI - Express delivery. PMID- 12894140 TI - Nurses disappearing from developing nations. PMID- 12894141 TI - Spain's scientists granted new career choice. PMID- 12894142 TI - Proteomics delivers on promise of cancer biomarkers. PMID- 12894143 TI - 'Emancipated' biotechs offer new drugs for neurodegeneration. PMID- 12894144 TI - Europe's last research chimps to retire. PMID- 12894145 TI - Technology, tradition unite in India's drug discovery scheme. PMID- 12894146 TI - Japan launches controversial Biobank project. PMID- 12894147 TI - US ponders law on genetic discrimination. PMID- 12894148 TI - Ethnicity puts clinical trials to the test. PMID- 12894150 TI - Govindarajan Padmanaban. PMID- 12894151 TI - KSHV reactivation in post-transplant Kaposi sarcoma. PMID- 12894156 TI - Coaxing coagulation with RNA and cell fragments. PMID- 12894157 TI - Sealing the leak, healing the heart. PMID- 12894158 TI - Seizing hold of seizures. PMID- 12894159 TI - Anthrax delivers a lethal blow to host immunity. PMID- 12894160 TI - Skipping to new gene therapies for muscular dystrophy. PMID- 12894161 TI - Muscular dystrophy meets the mesangioblast. PMID- 12894162 TI - Road map to metastasis. PMID- 12894163 TI - Cadmium's disguise dupes the estrogen receptor. PMID- 12894165 TI - Attacking pain at its source: new perspectives on opioids. AB - The treatment of severe pain with opioids has thus far been limited by their unwanted central side effects. Recent research promises new approaches, including opioid analgesics acting outside the central nervous system, targeting of opioid peptide-containing immune cells to peripheral damaged tissue, and gene transfer to enhance opioid production at sites of injury. PMID- 12894167 TI - Targeted movement of cell end factors in fission yeast. AB - Kinesins are microtubule-based motor proteins that transport cargo to specific locations within the cell. However, the mechanisms by which cargoes are directed to specific cellular locations have remained elusive. Here, we investigated the in vivo movement of the Schizosaccharomyces pombe kinesin Tea2 to establish how it is targeted to microtubule tips and cell ends. Tea2 is loaded onto microtubules in the middle of the cell, in close proximity to the nucleus, and then travels using its intrinsic motor activity primarily at the tips of polymerizing microtubules. The microtubule-associated protein Mal3, an EB1 homologue, is required for loading and/or processivity of Tea2 and this function can be substituted by human EB1. In addition, the cell-end marker Tea1 is required to anchor Tea2 to cell ends. Movement of Tea1 and the CLIP170 homologue Tip1 to cell ends is abolished in Tea2 rigor (ATPase) mutants. We propose that microtubule-based transport from the vicinity of the nucleus to cell ends can be precisely regulated, with Mal3 required for loading/processivity, Tea2 for movement and Tea1 for cell-end anchoring. PMID- 12894168 TI - Making impact. PMID- 12894170 TI - Why do worms need cholesterol? AB - Cholesterol is a structural component of animal membranes that influences fluidity, permeability and formation of lipid microdomains. It is also a precursor to signalling molecules, including mammalian steroid hormones and insect ecdysones. The nematode Caenorhabditis elegans requires too little cholesterol for it to have a major role in membrane structure. Instead, its most probable signalling functions are to control molting and induce a specialized non feeding larval stage, although no cholesterol-derived signalling molecule has yet been identified for these or any other functions. PMID- 12894171 TI - Cdk5 is a dynamo at the synapse. PMID- 12894172 TI - New dimensions in cell migration. PMID- 12894173 TI - Getting a GPR on spindle asymmetry. PMID- 12894174 TI - Actin puts on the squeeze. PMID- 12894175 TI - Talin forges the links between integrins and actin. PMID- 12894176 TI - Curling with kinesin. PMID- 12894177 TI - The sirens' call. PMID- 12894179 TI - A piece of history. PMID- 12894180 TI - Number crunching. PMID- 12894181 TI - Public markets show signs of life. PMID- 12894182 TI - US courts narrow patent exemptions. PMID- 12894183 TI - GM labeling in China beset by problems. PMID- 12894184 TI - Illumina tussles in court with ABI. PMID- 12894185 TI - US government previews biotech survey. PMID- 12894186 TI - EU directive on clinical trials penalizes small sponsors. PMID- 12894188 TI - Rediscovering plant-based drugs. PMID- 12894189 TI - Hybrid vigor. PMID- 12894190 TI - Data integration--connecting the dots. PMID- 12894191 TI - Genomic databases yield novel bioplastic producers. PMID- 12894192 TI - Biotechnology partnerships--medicine for an ailing industry? PMID- 12894193 TI - The science and politics of plant biotechnology--a personal perspective. PMID- 12894194 TI - Rethinking US leadership in food biotechnology. PMID- 12894195 TI - The stampede to convertibles. PMID- 12894197 TI - Screening for content--the evolution of high throughput. PMID- 12894198 TI - Biopharmaceutical benchmarks--2003. PMID- 12894199 TI - Thrombi--beware of red cells bearing gifts. PMID- 12894200 TI - Rise of the nanomachines. PMID- 12894201 TI - Constructing dwarf rice. PMID- 12894202 TI - Plant metabolic profiling en route to destination. PMID- 12894204 TI - Chemical discovery and global gene expression analysis in zebrafish. AB - The zebrafish (Danio rerio) provides an excellent model for studying vertebrate development and human disease because of its ex utero, optically transparent embryogenesis and amenability to in vivo manipulation. The rapid embryonic developmental cycle, large clutch sizes and ease of maintenance at large numbers also add to the appeal of this species. Considerable genomic data has recently become publicly available that is aiding the construction of zebrafish microarrays, thus permitting global gene expression analysis. The zebrafish is also suitable for chemical genomics, in part as a result of the permeability of its embryos to small molecules and consequent avoidance of external confounding maternal effects. Finally, there is increasing characterization and analysis of zebrafish models of human disease. Thus, the zebrafish offers a high-quality, high-throughput bioassay tool for determining the biological effect of small molecules as well as for dissecting biological pathways. PMID- 12894207 TI - Is the viability of the Lilly doctrine on the decline? PMID- 12894211 TI - The learning curve. PMID- 12894212 TI - Exclusion of germ plasm proteins from somatic lineages by cullin-dependent degradation. AB - In many animals, establishment of the germ line depends on segregation of a specialized cytoplasm, or 'germ plasm', to a small number of germline precursor cells during early embryogenesis. Germ plasm asymmetry involves targeting of RNAs and proteins to a specific region of the oocyte and/or embryo. Here we demonstrate that germ plasm asymmetry also depends on degradation of germline proteins in non-germline (somatic) cells. We show that five CCCH finger proteins, components of the Caenorhabditis elegans germ plasm, are targeted for degradation by the novel CCCH-finger-binding protein ZIF-1. ZIF-1 is a SOCS-box protein that interacts with the E3 ubiquitin ligase subunit elongin C. Elongin C, the cullin CUL-2, the ring finger protein RBX-1 and the E2 ubiquitin conjugation enzyme UBC5 (also known as LET-70) are all required in vivo for CCCH finger protein degradation. Degradation is activated in somatic cells by the redundant CCCH finger proteins MEX-5 and MEX-6, which are counteracted in the germ line by the PAR-1 kinase. We propose that segregation of the germ plasm involves both stabilization of germline proteins in the germ line and cullin-dependent degradation in the soma. PMID- 12894213 TI - RanGTP mediates nuclear pore complex assembly. AB - In metazoa, the nuclear envelope breaks down and reforms during each cell cycle. Nuclear pore complexes (NPCs), which serve as channels for transport between the nucleus and cytoplasm, assemble into the reforming nuclear envelope in a sequential process involving association of a subset of NPC proteins, nucleoporins, with chromatin followed by the formation of a closed nuclear envelope fenestrated by NPCs. How chromatin recruitment of nucleoporins and NPC assembly are regulated is unknown. Here we demonstrate that RanGTP production is required to dissociate nucleoporins Nup107, Nup153 and Nup358 from Importin beta, to target them to chromatin and to induce association between separate NPC subcomplexes. Additionally, either an excess of RanGTP or removal of Importin beta induces formation of NPC-containing membrane structures--annulate lamellae- both in vitro in the absence of chromatin and in vivo. Annulate lamellae formation is strongly and specifically inhibited by an excess of Importin beta. The data demonstrate that RanGTP triggers distinct steps of NPC assembly, and suggest a mechanism for the spatial restriction of NPC assembly to the surface of chromatin. PMID- 12894214 TI - Staphylococcus aureus alpha-toxin-induced cell death: predominant necrosis despite apoptotic caspase activation. AB - Recent data suggest that alpha-toxin, the major hemolysin of Staphylococcus aureus, induces cell death via the classical apoptotic pathway. Here we demonstrate, however, that although zVAD-fmk or overexpression of Bcl-2 completely abrogated caspase activation and internucleosomal DNA fragmentation, they did not significantly affect alpha-toxin-induced death of Jurkat T or MCF-7 breast carcinoma cells. Caspase inhibition had also no effect on alpha-toxin induced lactate dehydrogenase release and ATP depletion. Furthermore, whereas early assessment of apoptosis induction by CD95 resulted solely in the generation of cells positive for active caspases that were, however, not yet permeable for propidium iodide, a substantial proportion of alpha-toxin-treated cells were positive for both active caspases and PI. Finally, electron microscopy demonstrated that even in the presence of active caspases, alpha-toxin-treated cells displayed a necrotic morphology characterized by cell swelling and cytoplasmic vacuolation. Together, our data suggest that alpha-toxin-induced cell death proceeds even in the presence of activated caspases, at least partially, in a caspase-independent, necrotic-like manner. PMID- 12894215 TI - Decrease in intracellular superoxide sensitizes Bcl-2-overexpressing tumor cells to receptor and drug-induced apoptosis independent of the mitochondria. AB - At least two mechanisms of early cytosolic acidification during apoptotic signaling have been described, one that involves caspase 8 activation downstream of receptor ligation and another dependent on mitochondria-derived hydrogen peroxide during merocil-induced apoptosis. Here, we show that Bcl-2 inhibits both mechanisms of acidification. Moreover, Bcl-2 overexpression resulted in a slightly elevated constitutive level of superoxide anion and pH in CEM leukemia cells. Interestingly, decreasing intracellular superoxide concentration with an inhibitor of the beta-nicotinamide adenine dinucleotide phosphate oxidase or by transient transfection with a dominant-negative form of the guanosine triphosphate-binding protein Rac1 resulted in a significant increase in the sensitivity of CEM/Bcl-2 cells to CD95- or merocil-induced apoptosis. This increase in sensitivity was a direct result of a significant increase in caspase 8 activation and caspase 8-dependent acidification in the absence of caspase 9 activity or cytochrome c release. These findings suggest a mechanism of switching from mitochondria-dependent to mitochondria-independent death signaling in the same cell, provided the intracellular milieu is permissive for upstream caspase 8 activation, and could have implications for favorably tailoring tumor cells for drug treatment even when the mitochondrial pathway is compromised by Bcl-2. PMID- 12894216 TI - Unique structural features of a BCL-2 family protein CED-9 and biophysical characterization of CED-9/EGL-1 interactions. AB - The interactions between B-cell lymphoma 2 (BCL-2) family members are known to be mediated through the binding of the BH3 domain of a proapoptotic member to the BH3-binding groove of an antiapoptotic member. We determined the crystal structure of antiapoptotic CED-9, which reveals a unique C-terminal helix altering the common BH3-binding region. A coexpression system to produce CED-9 in complex with proapoptotic EGL-1 enabled us to show that the binding of EGL-1 to CED-9 is extremely stable, raising the melting temperature (T(M)) of CED-9 by 25 degrees C, and that the binding surface of CED-9 extends beyond the BH3-binding region and reaches the BH4 domain. Consistently, the T(M) and a 1H-15N correlation NMR spectrum of CED-9 in complex with EGL-1 are drastically different from those of CED-9 in complex with the EGL-1 BH3 peptide. The data suggest that the recognition between other BCL-2 family members may also involve much wider protein surfaces than is previously thought. PMID- 12894217 TI - CD95 ligand--death factor and costimulatory molecule? AB - The CD95 ligand is involved as a death factor in the regulation of activation induced cell death, establishment of immune privilege and tumor cell survival. In addition, CD95L may serve as a costimulatory molecule for T-cell activation. Alterations in expression or shedding of membrane and soluble CD95L are associated with numerous diseases, and underscore the pathophysiological relevance of the CD95/CD95L system. In most cases, the causal link between altered CD95L expression and pathophysiology is unknown. Given the potency of the molecule to regulate death and survival of many different cell types, the control of CD95L production, transport, storage, shedding and inactivation is of tremendous biological and clinical interest. This review summarizes the current knowledge, hypotheses and controversies about CD95L as a multifunctional ligand and receptor. It considers the different roles of membrane and soluble forms of CD95L and the complex networks of intracellular dynamics of protein trafficking, as well as the potential bidirectional signal transduction capacity of CD95L, with a focus on molecular interactions that have been worked out over the past years. PMID- 12894218 TI - Proapoptotic Bid binds to monolysocardiolipin, a new molecular connection between mitochondrial membranes and cell death. AB - Recent evidence indicates that the mitochondrial lipid cardiolipin may be instrumental in the proapoptotic action of Bcl-2 family proteins on mitochondrial membranes, leading to the release of apoptogenic factors. However, contrasting evidence indicates that progressive loss of cardiolipin occurs during apoptosis. Here we show that Bid, a crucial proapoptotic protein that integrates the action of other Bcl-2 family members, exhibits discrete specificity for metabolites of cardiolipin, especially monolysocardiolipin (MCL). MCL, normally present in the remodelling of mitochondrial lipids, progressively increases in mitochondria during Fas-mediated apoptosis as a by-product of cardiolipin degradation, and also enhances Bid binding to membranes. MCL may thus play a crucial role in connecting lipid metabolism, relocation of Bid to mitochondria and integrated action of Bcl-2 proteins on mitochondrial membranes. We propose that Bid interaction with MCL 'primes' the mitochondrial outer membrane via segregation of lipid domains, facilitating membrane discontinuity and leakage of apoptogenic factors. PMID- 12894219 TI - Retinoblastoma (Rb) protein upregulates expression of the Ifi202 gene encoding an interferon-inducible negative regulator of cell growth. AB - Studies have indicated that ectopic expression of p202, an interferon (IFN) inducible retinoblastoma (Rb)-binding protein, in cultured cells retards cell proliferation and modulates cell survival. Consistent with a role of p202 in cell cycle regulation, levels of p202 increase in cells arrested in the G0/G1 phase of cell cycle after withdrawal of serum growth factors. However, a role for p202 in cell growth arrest remains to be defined. Moreover, it remains unclear how levels of p202 are upregulated during the cell growth arrest. Here, we report that Rb upregulates expression of Ifi202 gene. We found that basal as well as IFN-induced levels of p202 were significantly higher in wild-type (Rb(+/+)) mouse embryonic fibroblasts (MEFs) than isogenic Rb(-/-) MEFs. Consistent with the regulation of Ifi202 gene by Rb, expression of functional Rb, but not a pocket mutant of it, stimulated the activity of a reporter whose expression was driven by the 5' regulatory region of Ifi202 gene. Importantly, the stimulation by Rb was dependent, in part, on a JunD/AP-1 DNA-binding site present in the 5'-regulatory region of the Ifi202 gene. Moreover, basal levels of p202 were significantly higher in wild-type (JunD(+/+)) than isogenic JunD(-/-) MEFs. Additionally, we found that increased expression of p202 potentiated the Rb-mediated inhibition of cell growth and mutations in the Rb-binding motif (LxCxE) of p202 significantly reduced cell survival. Together, our observations support the idea that the transcriptional activation of Ifi202 gene by Rb/JunD may be important for the regulation of cell growth and survival. PMID- 12894220 TI - Cell size regulation by the human TSC tumor suppressor proteins depends on PI3K and FKBP38. AB - TSC1 and TSC2 are responsible for the tumor suppressor gene syndrome tuberous sclerosis (TSC). Mammalian TSC genes have been shown to be involved in cell cycle regulation. Recently, in Drosophila, these data have been confirmed and TSC genes have further been demonstrated to affect cell size control. Here we provide supporting data for the fact that the latter function is conserved in mammals. Human TSC1 and TSC2 trigger mammalian cell size reduction and a dominant-negative TSC2 mutant induces increased size. These effects occur in all cell cycle phases, are dependent on the activity of the phosphoinositide-3-kinase and are abolished by co-overexpression of a dominant-negative Akt mutant. Two independent naturally occurring and disease-causing mutations within the TSC2 gene eliminate tuberin's capacity to affect cell size control, emphasizing the relevance of this function for the development of the disease. The same mutations have earlier been shown not to affect tuberin's antiproliferative capacity. That the consequences of modulated TSC gene expression on cell proliferation and on cell size can be assigned to separable functions is further supported by two findings: A mutation within the TSC1 gene, earlier shown to still harbor anti-proliferative effects, was found to eliminate the cell size regulating functions. An important mammalian cell size regulator, c-Myc, was found to inhibit tuberin's antiproliferative capacity, but to have no effects on tuberin-dependent cell size control. To obtain further mechanistical insights, microarray screens for genes involved in TSC1- or TSC2-mediated cell size effects were performed. Antisense experiments revealed that the so observed regulation of the FK506-binding protein, FKBP38, plays a role in TSC gene-dependent cell size regulation. These data provide new insights into mammalian cell size regulation and allow a better understanding of the function of human TSC genes. PMID- 12894221 TI - CrkIII: a novel and biologically distinct member of the Crk family of adaptor proteins. AB - As the role for adaptor proteins constantly proliferates, appreciation of their importance has never been higher. The Crk family of adaptor proteins is no exception. Currently comprising four members, v-Crk, CrkI, CrkII and Crk-like protein, we have introduced a fifth member, CrkIII. Cloned by the CORT technique, CrkIII is identical in sequence to CrkII until the second of its two SH3 domains, which is disrupted partway through and results in a nonfunctional domain and a unique C-terminal sequence. We have demonstrated the existence of native CrkIII at the message level using RT-PCR and RNAse protection assays, and at the protein level in mouse fibroblasts. We show that CrkII overexpression is capable of enhancing insulin-stimulated ERK activity, whereas CrkIII is not, thus partially characterizing a novel member of the Crk family and elucidating important effects mediated by the c-terminal SH3 domain. PMID- 12894222 TI - Inhibition of MMTV transcription by HDAC inhibitors occurs independent of changes in chromatin remodeling and increased histone acetylation. AB - Increased histone acetylation has been associated with activated gene transcription and decreased acetylation with repression. However, there is a growing number of genes known, which are downregulated by histone deacetylase (HDAC) inhibitors through unknown mechanisms. This study examines the mechanism by which the mouse mammary tumor virus (MMTV) promoter is repressed by the HDAC inhibitor, trichostatin A (TSA). We find that this repression is transcriptional in nature and that it occurs in the presence and absence of glucocorticoids. TSA decreases MMTV transcription at a rapid rate, reaching maximum in 30-60 min. In contrast with previous reports, the repression does not correlate with an inhibition of glucocorticoid-induced nuclease hypersensitivity or NF1-binding at the MMTV promoter. Surprisingly, TSA does not induce sizable increases in histone acetylation at the MMTV promoter nor does it inhibit histone deacetylation, which accompanies deactivation of the glucocorticoid-activated MMTV promoter. Repression of MMTV transcription by TSA does not depend on the chromatin organization of the promoter because a transiently transfected MMTV promoter construct with a disorganized nucleoprotein structure was also repressed by TSA treatment. Mutational analysis of the MMTV promoter indicates that repression by TSA is mediated through the TATA box region. These results suggest a novel mechanism that involves acetylation of nonhistone proteins necessary for basal transcription. PMID- 12894223 TI - Proteomic analysis of ubiquitin-proteasome effects: insight into the function of eukaryotic initiation factor 5A. AB - The global effect of ubiquitin-proteasome (UP) inhibitors on leukemic cell proteome was analysed. A total of 39 protein spots, affected by UP inhibitors, were identified, including 11 new apoptosis-associated proteins. They are involved in different cellular functions and four were associated with caspase-3 activation. Eukaryotic initiation factor 5A (eIF-5A) was identified in two spots; however, the peptide mass-fingerprinting for the accumulated one included a peptide with lysine50, indicating that hypusine formation was suppressed during UP inhibitor-induced apoptosis. Hypusine modification ensues immediately following translation of eIF-5A precursor, unless cells are treated with the modification inhibitors diaminoheptane. However, UP inhibitors induced a much stronger accumulation of unmodified eIF-5A compared to the effect of diaminoheptane. We further showed the unmodified eIF-5A was regulated in a proteasome-dependent manner. Inhibition of hypusine formation by diaminoheptane triggered apoptosis, but of particular interest is the finding that eIF-5A expression inhibition by antisense oligodeoxynucleotides significantly enhanced the stimulating effect of GM-CSF on cell growth. Therefore, the eIF-5A accumulation played important roles in the apoptosis induced by UP inhibitors. Moreover, hypusine inhibition in apoptosis was further revealed to be associated with the subcellular localization of eIF-5A. Our data pave the way to a better understanding of the mechanisms by which UP system has been linked to apoptosis. PMID- 12894224 TI - Role of IFI 16, a member of the interferon-inducible p200-protein family, in prostate epithelial cellular senescence. AB - Recent studies have implicated interferon signaling in the regulation of cellular senescence. However, the role of specific interferon-inducible proteins in cellular senescence remains to be defined. Here we report that IFI 16, an interferon-inducible transcriptional modulator from the p200-protein family, contributes to cellular senescence of prostate epithelial cells. Normal human prostate epithelial cells (PrEC) in culture expressed detectable levels of IFI 16, and the levels increased more than fourfold when cells approached cellular senescence. Consistent with a role of IFI 16 in cellular senescence, human prostate cancer cell lines either did not express IFI 16 or expressed a variant form, which was primarily detected in the cytoplasm of prostate cancer cells and not in the nucleus. Moreover, overexpression of functional IFI 16 in human prostate cancer cell lines inhibited colony formation. Additionally, ectopic expression of IFI 16 in clonal prostate cancer cell lines was associated with a senescence-like phenotype, production of senescence-associated beta-galactosidase (a biochemical marker for cellular senescence), and reduction of S-phase cells in culture. Importantly, upregulation of p21WAF1 and inhibition of E2F-stimulated transcription accompanied inhibition of cell growth by IFI 16 in prostate cancer cell lines. Collectively, our observations support the idea that increased levels of IFI 16 in PrECs contribute to senescence-associated irreversible cell growth arrest. PMID- 12894225 TI - p38 MAP kinase modulates Smad-dependent changes in human prostate cell adhesion. AB - Transforming growth factor beta (TGFbeta) regulates cell adhesion, proliferation, and differentiation in a variety of cells. Smad proteins are receptor-activated transcription factors that translocate to the nucleus in response to TGFbeta. We demonstrate here that TGFbeta increases cell adhesion in metastatic PC3-M prostate cancer cells. TGFbeta treatment of PC3-M cells leads to nuclear translocation of R-Smad proteins. We show that Smad proteins are necessary, but not sufficient, for TGFbeta-mediated cell adhesion. After showing that TGFbeta upregulated p38 MAP kinase activity in PC3-M cells, we show that inhibition of p38 MAP kinase partially blocked TGFbeta-mediated increase in cell adhesion, as well as nuclear translocation of Smad3. Finally, we show that Smad3 is phosphorylated by p38 MAP kinase in vitro. These findings implicate crosstalk between the MAP kinase and Smad signaling pathways in TGFbeta's regulation of cell adhesion in human prostate cells. This represents a mechanism by which the pleiotropic effects of TGFbeta may be channeled to modulate cell adhesion. PMID- 12894226 TI - Role of p53 and NF-kappaB in epigallocatechin-3-gallate-induced apoptosis of LNCaP cells. AB - We have recently shown that oral consumption of green tea polyphenols inhibits prostate carcinogenesis in transgenic mouse model of prostate cancer and suggested that induction of apoptosis in prostate cancer cells is responsible for these effects. Much of the chemopreventive effects of green tea are attributed to its major polyphenolic constituent (-) epigallocatechin-3-gallate (EGCG). In the present study, we report that EGCG-induced apoptosis in human prostate carcinoma LNCaP cells is mediated via modulation of two related pathways: (a) stabilization of p53 by phosphorylation on critical serine residues and p14ARF-mediated downregulation of murine double minute 2(MDM2) protein, and (b) negative regulation of NF-kappaB activity, thereby decreasing the expression of the proapoptotic protein Bcl-2. EGCG-induced stabilization of p53 caused an upregulation in its transcriptional activity, thereby resulting in activation of its downstream targets p21/WAF1 and Bax. Thus, EGCG had a concurrent effect on two important transcription factors p53 and NF-kappaB, causing a change in the ratio of Bax/Bcl-2 in a manner that favors apoptosis. This altered expression of Bcl-2 family members triggered the activation of initiator capsases 9 and 8 followed by activation of effector caspase 3. Activation of the caspases was followed by poly (ADP-ribose) polymerase cleavage and induction of apoptosis. Taken together, the data indicate that EGCG induces apoptosis in human prostate carcinoma cells by shifting the balance between pro- and antiapoptotic proteins in favor of apoptosis. PMID- 12894227 TI - Eiger and its receptor, Wengen, comprise a TNF-like system in Drosophila. AB - In mammals, members of the tumor necrosis factor (TNF) family play an important role in the regulation of cellular proliferation, differentiation and programmed cell death. We describe isolation and characterization of an orthologous ligand/receptor axis in Drosophila. The ligand, designated Eiger, is a type II membrane glycosylated protein, which can be cleaved at residue 145 and released from the cell surface as a soluble factor, thereby representing the first potential cytokine to be described in Drosophila. Eiger exists in two alternatively spliced isoforms, Eiger long (Eiger-L) and Eiger short (Eiger-s), both of which are expressed throughout development and in the adult. We also describe the isolation and characterization of a novel Drosophila member of the TNF receptor family, designated Wengen, which is a type I membrane protein that can physically interact with the recently described TRAF2 homolog dTRAF2. Both Eiger and Wengen are expressed in distinctive patterns during embryogenesis and Eiger is responsive to genotoxic stress. Forced expression of Eiger-L, Eiger-s or Wengen, caused apoptotic cell death which could be rescued by caspase inhibitors or the JNK phosphatase Puckered. In addition, Eiger-induced cell killing was attenuated by RNAi-mediated suppression of Wengen. Our results illustrate that Eiger and Wengen represent proximal components of an evolutionarily conserved TNF like signaling pathway in Drosophila. PMID- 12894228 TI - Regulation of NF-kappaB2/p100 processing by its nuclear shuttling. AB - Processing of the NF-kappaB2 precursor protein p100 to generate p52 is an important step of NF-kappaB regulation. This proteolytic event is tightly regulated by sequences located at the C-terminal portion of p100. Constitutive processing of p100 occurs in certain lymphoma cells due to the loss of its C terminal regulatory domain, although the underlying mechanisms remain unknown. We show here that the constitutive processing of C-terminal truncation mutants of p100 is associated with their active nuclear translocation. Deletion of the C terminal death domain of p100 triggers a low, but significant, level of nuclear translocation and processing. Disruption of the ankyrin-repeat domain of p100 further enhances its nuclear shuttling activity, which is again associated with elevated level of processing. More importantly, mutation of the nuclear localization signal (NLS) of p100 abolishes its processing, and this defect can be rescued by fusion of a heterologous NLS to the amino- or carboxyl-terminus of the p100 mutant. These results suggest that nuclear shuttling is a mechanism regulating the processing of NF-kappaB2/p100. PMID- 12894229 TI - Oestrogens and selective oestrogen receptor (ER) modulators regulate EGF receptor gene expression through human ER alpha and beta subtypes via an Sp1 site. AB - Through the analysis of the transient expression of the luciferase reporter gene in HeLa cells, an evaluation has been made of the transcriptional activity of oestrogens and of selective oestrogen receptor (ER) modulators (SERMs), mediated by the alpha and beta isoforms of the ER, on the epidermal growth factor receptor gene promoter. Oestrogen-activated ERbeta presents a lower transcriptional activity compared with ERalpha, probably due to structural differences in the AF 1 regions of the receptors. Also SERMs induce different responses depending on the receptor isoform bound. Indeed, the phyto-oestrogens, genistein and daidzein, act as weak agonists of the oestrogenic activity via ERalpha, but as full agonists when bound to ERbeta. The synthetic SERM 4OH-tamoxifen, on the other hand, displays an opposite behaviour since it exerts a full agonist action through ERalpha, but acts as a full antagonist via ERbeta. As we have previously shown for ERalpha, an ERbeta/Sp1 functional synergism has also been highlighted, by means of gel mobility shift assays. Moreover, our results show that the sensitivity of target tissues to oestrogens and SERMs can be affected by coexpression of ERs, depending on the formation of appropriate levels of homo- and heterodimers, thus providing a useful approach to predict the effects of hormonal treatment. PMID- 12894230 TI - Functional inactivation of Cdk9 through oligomerization chain reaction. AB - The oligomerization chain reaction (OCR) strategy is a recently described technique for inactivation of target proteins that function as homoassociate complexes. This novel strategy is based on the fusion of self-associating coiled coil (CC) domain of the nuclear factor promyelocytic leukemia (PML) to target proteins. Here, we present the successful application of the OCR strategy for inactivation of the heterodimeric Cdk9/cyclin T1 complex. Cyclin T1/Cdk9 (P-TEFb) complex is a positive regulator of gene transcription, whose function is underlined by the ability to phosphorylate the carboxyl-terminal domain (CTD) of the RNA polymerase II conferring productive transcript elongation. Fusion of the CC domain to Cdk9 leads to the formation of high molecular complexes to which the endogenous cyclin T1 is recruited. The CC-Cdk9 chimera effectively inhibits HIV-1 Tat activation, whose transcription activity is exquisitely dependent upon cyclin T1/Cdk9 function. Furthermore, expression of CC-Cdk9 protein inhibits cell proliferation, as shown by colony-formation assay. Collectively, our findings add further support to the OCR strategy for functional inactivation of hetero associated factors such as the Cdk9/cyclin T1 complex, and highlight a putative function of Cdk9 in cell growth control. PMID- 12894231 TI - Loss of expression, and mutations of Smad 2 and Smad 4 in human cervical cancer. AB - Mutations in Smads, intermediates of transforming growth factor-beta signaling, are known to contribute to the loss of sensitivity to transforming growth factor beta, a common feature of many neoplastic cells. However, not much information is available on Smad alterations in cervical cancer and so we probed, for the first time, for alterations in Smad 2 and Smad 4 genes using human cervical cancer cell lines and human cervical tissue samples. Using PCR/reverse transcription-PCR, single-stranded conformation polymorphism analysis and DNA sequencing, we observed a deletion of 'G' in the L3 loop (crucial in Smad-receptor interaction) in C-33A cells, and an insertion of 'A' in codon 122 (loss of MH2 domain) from a cervical tumor sample, both of which caused frame shift and pretermination in Smad 2. In addition, a G/A transition at 31 bp upstream-nontranslated regions of exon 8 of Smad 4 was found in Bu 25TK cells. Smad 2 expression was less in some of the cervical tumor samples than that of nonmalignant samples and six cancer samples showed C-terminal deletions that abolish Smad 2 phosphorylation sites. The loss of expression of Smad 4 found in some cervical tumor samples was due to transcription loss rather than deletion of the gene. Our results highlight an important role for Smad 2 and Smad 4 in human cervical tumors. PMID- 12894232 TI - Cyclin-E is a strong predictor of endocrine therapy failure in human breast cancer. AB - Recently, cyclin-E was reported to be the most prominent prognostic factor for breast cancer outcome described so far, even surpassing axillary nodal involvement. Earlier studies on the prognostic value of cyclin-E in breast cancer, however, yielded heterogeneous results. Therefore, we set out to confirm and extend these results by quantitative Taqman RT-PCR of cyclin-E levels in 277 resectable breast cancers. Cyclin-E levels were not associated with relapse-free survival (RFS) or overall survival (OS) in the total cohort of patients, or in the subset of patients without involved lymph nodes that were not treated with adjuvant systemic therapy. Besides several classical clinicopathological factors, the interaction between cyclin-E and adjuvant endocrine therapy (P=0.01, HR=3.04, 95% CI: 1.30-7.09) was found to contribute significantly in multivariate analyses. Cyclin-E levels were associated with poor RFS specifically in patients treated with adjuvant endocrine therapy (n=108, P=0.001, HR=4.01, 95% CI: 1.76 9.12), independent of estrogen receptor status. In conclusion, cyclin-E is not a pure prognostic factor in breast cancer, but rather a predictor of failure of endocrine therapy. Differences in literature on the presumed prognostic value of cyclin-E may be due to differences in treatment. Assessment of cyclin-E levels can aid in improving adjuvant treatment selection. PMID- 12894233 TI - The transformation suppressor protein Pdcd4 shuttles between nucleus and cytoplasm and binds RNA. AB - The Pdcd4 gene has originally been isolated in a search for genes that are activated in cells undergoing apoptosis. Independent of these studies, the Pdcd4 gene has been implicated in the suppression of tumor-promoter-mediated transformation of keratinocytes and as a downstream target of Myb in hematopoietic cells. The Pdcd4 protein has weak homology to the eucaryotic translation initiation factor eIF4G and has been shown to interact with certain translation initiation factors. To explore the molecular function of the Pdcd4 protein, we have studied its subcellular localization. We show that the Pdcd4 protein is a predominantly nuclear protein under normal growth conditions and that it is exported from the nucleus by a leptomycin B-sensitive mechanism upon serum withdrawal. The protein contains two nuclear export signals, one of which is very potent. In addition, we demonstrate that the Pdcd4 protein has RNA binding activity and that the sequences involved in RNA-binding are located in the amino-terminal part of the protein. Taken together, our data raise the possibility that Pdcd4 is involved in some aspect of nuclear RNA metabolism in addition to its suspected role in protein translation. PMID- 12894234 TI - Melanoma cells can tolerate high levels of transcriptionally active endogenous p53 but are sensitive to retrovirus-transduced p53. AB - Malignant melanomas are frequently characterized by elevated levels of wild-type p53, suggesting that p53 function could be suppressed by a mechanism different from p53 mutation. We analysed the functionality of the p53-signaling pathway in a panel of seven human melanoma cell lines consisting of one p53-deficient line, two lines with mutant p53, and four lines expressing wild-type p53. Only lines with wild-type p53 were characterized by elevated levels of endogenous p21, high activity of p53-responsive reporters and accumulation of p53 in response to genotoxic stress, common properties of functional p53. The presence of wild-type p53 was associated with depletion or loss of p14ARF and p16 expression. The levels of p33ING1b and p24ING1c, two major products of Ing1 locus and putative coregulators of p53, were elevated in all cell lines tested; however, ectopic expression of either ING1 isoform had no effect on cell proliferation. All lines retained expression of Apaf-1, and all but one remained sensitive to ectopic expression of retrovirus-transduced p53. Our data indicate that regardless of abnormally high levels of p53 in melanomas, their p53 remains competent in transactivation of its targets, and, if highly overexpressed, capable of growth inhibition. Hence, the p53 pathway in malignant melanomas can be considered for pharmacological targeting and anticancer gene therapy. PMID- 12894235 TI - Gene expression profiling identifies molecular subtypes of gliomas. AB - Identification of distinct molecular subtypes is a critical challenge for cancer biology. In this study, we used Affymetrix high-density oligonucleotide arrays to identify the global gene expression signatures associated with gliomas of different types and grades. Here, we show that the global transcriptional profiles of gliomas of different types and grades are distinct from each other and from the normal brain. To determine whether our data could be used to uncover molecular subtypes without prior knowledge of pathologic type and grade, we performed K-means clustering analysis and found evidence for three clusters with the aid of multidimensional scaling plots. These clusters corresponded to glioblastomas, lower grade astrocytomas and oligodendrogliomas (P<0.00001). A predictor constructed from the 170 genes that are most differentially expressed between the subsets correctly identified the type and grade of all samples, indicating that a relatively small number of genes can be used to distinguish between these molecular subtypes. These results further define molecular subsets of gliomas which may potentially be used for patient stratification, and suggest potential targets for treatment. PMID- 12894236 TI - Microarray analysis uncovers retinoid targets in human bronchial epithelial cells. AB - Retinoids, the natural and synthetic derivatives of vitamin A, have a role in cancer treatment and prevention. There is a need to reveal mechanisms that account for retinoid response or resistance. This study identified candidate all trans-retinoic acid (RA) target genes linked to growth suppression in BEAS-2B human bronchial epithelial cells. Microarray analyses were performed using Affymetrix arrays. A total of 11 RA-induced species were validated by reverse transcription polymerase chain reaction (RT-PCR), Western or Northern analyses. Three of these species were novel candidate RA-target genes in human bronchial epithelial cells. These included: placental bone morphogenetic protein (PLAB), polyamine oxidase isoform 1 (PAOh1) and E74-like factor 3 (ELF3). Expression patterns were studied in RA-resistant BEAS-2B-R1 cells. In BEAS-2B-R1 cells, RA dysregulated the expression of the putative lymphocyte G0/G1 switch gene (G0S2), heme oxygenase 1 (HMOX1), tumor necrosis factor-alpha-induced protein 2 (TNFAIP2), inhibitor of DNA binding 1(Id1), fos-like antigen 1 (FOSL1), transglutaminase 2 (TGM2), asparagine synthetase (ASNS), PLAB, PAOh1 and ELF3, while prominent induction of insulin-like growth-factor-binding protein 6 (IGFBP6) still occurred. In summary, this study identified 11 candidate RA-target genes in human bronchial epithelial cells including three novel species. Expression studies in BEAS-2B-R1 cells indicated that several were directly implicated in RA signaling, since their aberrant expression was linked to RA resistance of human bronchial epithelial cells. PMID- 12894237 TI - Early central nervous system evolution: an era of skin brains? PMID- 12894238 TI - PEDF: a multifaceted neurotrophic factor. PMID- 12894239 TI - Prefrontal and medial temporal lobe interactions in long-term memory. PMID- 12894240 TI - A clockwork web: circadian timing in brain and periphery, in health and disease. PMID- 12894241 TI - Spontaneous confabulation and the adaptation of thought to ongoing reality. PMID- 12894242 TI - The biochemistry of neuronal necrosis: rogue biology? PMID- 12894243 TI - Can animals recall the past and plan for the future? AB - According to the 'mental time travel hypothesis' animals, unlike humans, cannot mentally travel backwards in time to recollect specific past events (episodic memory) or forwards to anticipate future needs (future planning). Until recently, there was little evidence in animals for either ability. Experiments on memory in food-caching birds, however, question this assumption by showing that western scrub-jays form integrated, flexible, trial-unique memories of what they hid, where and when. Moreover, these birds can adjust their caching behaviour in anticipation of future needs. We suggest that some animals have elements of both episodic-like memory and future planning. PMID- 12894244 TI - The genetics of malignant melanoma: lessons from mouse and man. AB - Therapeutic resistance and proclivity for metastasis are hallmarks of malignant melanoma. Genetic, epidemiological and genomic investigations are uncovering the spectrum of stereotypical mutations that are associated with melanoma and how these mutations relate to risk factors such as ultraviolet exposure. The ability to validate the pathogenetic relevance of these mutations in the mouse, coupled with advances in rational drug design, has generated optimism for the development of effective prevention programmes, diagnostic measures and targeted therapeutics in the near future. PMID- 12894245 TI - Caveolae: mining little caves for new cancer targets. AB - Caveolae exist at cell surfaces as caveolin-coated invaginations that perform transport and signalling functions influencing cell growth, apoptosis, angiogenesis and transvascular exchange. Caveolin could constitute a key switch in tumour development through its function as a tumour suppressor and as a promoter of metastasis, chemoresistance and survival. Targeting of drugs and gene vectors to tissue-specific proteins in caveolae allows selective delivery into vascular endothelial cells in vivo and might even improve direct access to solid tumour cells. Therefore, caveolae seem to be rich in potential targets for cancer imaging and therapeutics. PMID- 12894246 TI - The emerging role of lysophosphatidic acid in cancer. AB - The bioactive phospholipid lysophosphatidic acid (LPA) stimulates cell proliferation, migration and survival by acting on its cognate G-protein-coupled receptors. Aberrant LPA production, receptor expression and signalling probably contribute to cancer initiation, progression and metastasis. The recent identification of ecto-enzymes that mediate the production and degradation of LPA, as well as the development of receptor-selective analogues, indicate mechanisms by which LPA production or action could be modulated for cancer therapy. PMID- 12894247 TI - Control of gut differentiation and intestinal-type gastric carcinogenesis. AB - Gastric cancer is one of the world's most common cancers. Its carcinogenic pathway is mainly associated with Helicobacter pylori infection, subsequent inflammation and tissue regeneration. During the regeneration process, cells deviate from the normal pathway of gastric differentiation to an 'intestinal phenotype', which is thought to be precancerous and associated with the intestinal type of gastric cancer. Inappropriate activation of intestine-specific transcription factors could contribute to the occurrence of the intestinal-type cancer of the stomach. PMID- 12894248 TI - Chemoprevention of colon cancer by calcium, vitamin D and folate: molecular mechanisms. AB - Recent findings have indicated that dietary calcium, vitamin D and folate can modulate and inhibit colon carcinogenesis. Supporting evidence has been obtained from a wide variety of preclinical experimental studies, epidemiological findings and a few human clinical trials. Important molecular events and cellular actions of these micronutrients that contribute to their tumour-modulating effects are discussed. They include a complex series of signalling events that affect the structural and functional organization of colon cells. PMID- 12894249 TI - Clinical development of gene therapy for colorectal cancer. AB - Colorectal cancer (CRC) is the second most common type of malignancy in Western nations. Improvements in surgical and radiotherapeutic techniques and the increased availability of new cytotoxic drugs have improved outcome, but 50% of patients still die from recurrent or metastatic disease. Several features of its natural history render CRC a good candidate for gene therapy. Techniques include gene replacement, virus-directed enzyme-prodrug therapy, immune manipulation and virotherapy, all of which have entered clinical trials. PMID- 12894250 TI - Telomere dysfunction and the initiation of genome instability. AB - Tumour growth is an evolutionary process that is characterized by the selection of clonal populations of cells that acquire distinct genetic changes. Many cancer therapies aim to exploit the specific changes that occur in cancer cells, but understanding the underlying mechanisms of genomic instability that cause these mutations could lead to more effective therapies. If common mechanisms exist for initiating genomic instability in tumours, selection could explain the differences in specific gene mutations that accumulate in different tumour types. The cause of genomic instability in human tumours is unclear, although there is evidence to indicate that telomere dysfunction could make an important contribution. PMID- 12894251 TI - Responding to 'nanotech' concerns. PMID- 12894252 TI - Large research consortia not the answer to European fragmentation. PMID- 12894253 TI - Intermolecular potentials--past, present, future. PMID- 12894255 TI - Magnetic nanostructures: Edge atoms do all the work. PMID- 12894256 TI - Silicon membranes: A microscale Brownian ratchet. PMID- 12894257 TI - Materials conservation: Not-so-new technology. PMID- 12894259 TI - The use of environmental scanning electron microscopy for imaging wet and insulating materials. AB - The environmental scanning electron microscope (ESEM) is a direct descendant of the conventional SEM, but also permits wet and insulating samples to be imaged without prior specimen preparation. A low pressure (up to around 10 torr) of a gas can be accommodated around the sample. When this gas is water, hydrated samples can be maintained in their native state. Whether the gas is water or some other gas, ions formed on collisions between electrons emitted from the sample and the gaseous molecules drift back towards the sample surface helping to reduce charge build up. This eliminates the need for insulators to be subjected to a conductive surface coating. These two key advantages of ESEM open up a wide range of materials to the power of scanning electron microscopy. PMID- 12894261 TI - Temporal lobe hypogenesis associated with arachnoid cyst in patients with epilepsy. AB - OBJECTIVE: To determine the frequency of temporal lobe hypogenesis (TLH) associated with arachnoid cysts (AC) in patients with epilepsy. METHOD: We retrospectively revised 655 consecutive MRI scans from patients followed in our epilepsy clinic. We identified patients with temporal AC and then performed careful visual analysis in a workstation. Patients with evident expansive or destructive lesions were excluded. RESULTS: Only 4 (0.6%) patients had AC in the left temporal lobe, all associated with TLH. In addition, there were also ipsilateral dysgenetic characteristics in the ipsilateral hippocampus including abnormal shape and axis, and hyperintense T2 signal. In one patient this hippocampal abnormality was bilateral. CONCLUSION: AC with TLH was rarely found in our patients with epilepsy and it was always associated with hippocampal dysgenesis. Although volumetric reduction of the temporal lobe can be observed in patients with epilepsy and hippocampal abnormalities, the presence of adjacent AC points to a malformative etiology. PMID- 12894262 TI - Is the prevalence of celiac disease increased among epileptic patients? AB - OBJECTIVE: To assess the prevalence of celiac disease (CD) among a group of epileptic patients attending the Epilepsy Clinics of two general hospitals in the city of Brasilia (DF), Brazil. METHOD: Serum samples were collected from 255 epileptic patients (119 children, 136 adults) originating from Epilepsy Clinics, and from a control group composed by 4405 individuals (2034 children, 2371 adults) attending the Laboratory of Clinical Analysis, for routine blood testing. The diagnosis of CD was determined by the antiendomysium antibody (IgA-EMA) test and by small intestine biopsy. RESULTS: two of the 255 epileptic patients (1:127) and fifteen subjects from the control group (1:293) tested positive for the IgA EMA assay. CONCLUSION: the prevalence of CD was 2.3 times higher in epileptic patients than in controls (7.84 per 1000 versus 3.41 per 1000). Although still not statistically significant, this result is highly suggestive of an increased prevalence of CD among epileptic patients. PMID- 12894263 TI - Attentional disorders in patients with complex partial epilepsy. AB - A study of concentrated attention patterns in epileptic patients was conducted with the objectives: characterization of the patients' epileptic condition; assessment of the concentrated attention levels in epileptic and nonepileptic individuals; comparison of the attention levels of the two groups. An evaluation was performed of 50 adult outpatients with complex partial seizures and 20 non epileptic individuals (comparative group) at the Neuroepilepsy Ambulatory Unit, State University of Campinas SP, Brazil. METHOD: characterization of seizure types, frequency and duration; concentrated attention assessment (Concentrated Attention Test - Toulouse-Pi ron); comparison of the epileptic with non-epileptic individuals. RESULTS: A statistically significant difference was observed between the groups with regard to Correct Response, Wrong Response and No Response. A difference was observed in relation to Time, but it was statistically insignificant. The epileptic patients presented inferior cognitive performance in relation to concentrated attention when compared with the non-epileptic individuals, findings compatible with the clinical complaints. PMID- 12894264 TI - Brazilian version of the Mattis dementia rating scale: diagnosis of mild dementia in Alzheimer's disease. AB - OBJECTIVES: To verify the diagnostic accuracy of the Brazilian version of the Mattis Dementia Rating Scale (DRS) in the diagnosis of patients with mild dementia in Alzheimer's disease (AD); to verify the interference of the variables age and schooling on the performance of the DRS. METHOD: The DRS was administered to 41 patients with mild AD and to 60 controls. In order to analyze the effects of age and schooling on the performance of the tests, patients and controls were separated into three age groups and three levels of schooling. RESULTS: The cutoff score of 122 showed a sensitivity of 91.7 % and specificity of 87.8 %. Age and schooling interfered in the DRS total score and in the scores of its subscales. CONCLUSION: The DRS showed good diagnostic accuracy in the discrimination of patients with mild AD from the control individuals. In the sample examined, the effects of schooling were more marked than age. PMID- 12894265 TI - The expression of NGFr and PGP 9.5 in leprosy reactional cutaneous lesions: an assessment of the nerve fiber status using immunostaining. AB - The effects of reactional episodes on the cutaneous nerve fibers of leprosy patients was assessed in six patients (three with reversal reactions and three with erythema nodosum leprosum). Cryosections of cutaneous biopsy of reactional lesions taken during the episode and of another sample during the remission period were immunostained with anti-NGFr and anti-PGP 9.5 (indirect immunofluorescence). We found no significant statistical difference in the number of NGFr- and PGP 9.5-positive fibers between the reactional and post-reactional groups. A significant difference was detected between the number of NGFr and PGP 9.5-stained fibers inside of the reactional group of biopsy cryosections but this difference was ascribed to the distinct aspects of the nerve fibers displayed whether stained with anti-NGFr or with anti-PGP 9.5; NGFr-positive branches looked larger and so interpreted as containing more fibers. In addition, a substantial number NGFr-positive fibers were PGP 9.5-negative. No differences in the number of stained fibers among the distinct cutaneous regions examined (epidermis + upper dermis, mid and deep dermis) was detected. In conclusion, the number of PGP- and NGFr-positive fibers were not significantly different in the reactional and post-reactional biopsies in the present study. NGFr-staining of the nerve fibers is different from their PGP-imunoreactivity and the evaluation of the nerve fiber status on an innervated target organ should be carried out choosing markers for both components of nerve fibers (Schwann cells and axons). PMID- 12894266 TI - Clinical and cerebrospinal fluid (CSF) profile and CSF criteria for the diagnosis of spinal cord schistosomiasis. AB - OBJECTIVES: To describe the clinical and CSF findings among patients with presumptive neuroschistosomiasis (NS) and to suggest a classification for the CSF diagnosis of presumptive NS. METHOD: The charts of all patients whose CSF exam was performed at the CSF Lab, Jos Silveira Foundation, Salvador, Brazil, from 1988 to 2002 were reviewed. Those with clinically suspected NS whose indirect fluorescent antibody test (IFA) and or hemagglutination-inhibiting antibodies test (HAI) were positive to S. mansoni were identified. RESULTS: Of 377 patients, 67.9% were males; the median age was 36 years (mean 37 + 16 yrs, range 3-82 yrs). The most frequent complaints were paraparesis (59.9%), urinary retention (36.2%), lower limb pain (22.8%). WBC of CSF (count/mm ) was > 4 in 66.0% (mean 83 + 124, median 40, range 4.3-1,100), protein (mg/dl) was > 40 in 84.6% (mean 185 + 519, median 81, range 41-6,800) and eosinophils were present in 46.9%. IFA and HAI were positive in 75.3%. WBC > 4 and presence of eosinophils were associated with IFA and HAI positive (67.3% versus 51.4%, p 0.014; 49.1% versus 23.0%, p 0.0001, respectively) and protein > 40 was not (85.4% versus 77.0%, p 0.09). Presence of WBC > 4, protein > 40 and eosinophils was associated with IFA and HAI positive (71.6% versus 38.2%, p 0.0003) but presence of eosinophils and any other combination of WBC and protein were not. CONCLUSION: NS should be considered as a possible diagnosis in patients who had had contact with schistosome-infected water and present with spinal cord compromising. Presence of IFA and HAI positive to S. mansoni, WBC > 4, protein > 40 and presence of eosinophils in the CSF may be considered as a criterium of highly probable presumptive diagnosis. PMID- 12894267 TI - Urinary symptoms in Parkinson's disease: prevalence and associated factors. AB - The authors present a cross-sectional study involving 61 patients with idiopathic Parkinson's disease (PD) who were consecutively examined and compared to a control group with 74 subjects. Only patients who fulfilled the standard diagnostic criteria for PD and whose brain magnetic resonance imaging was normal were included. The objective of the study was to evaluate the prevalence of inferior urinary tract symptoms in PD and to study the possible association between clinical factors to urinary dysfunction. ln the patient group, 39.3% presented urinary symptoms when compared to 10.8% in the control group. All symptomatic patients presented irritative symptoms. The most common irritative symptom PD was nocturia, followed by frequency and urinary incontinence. Around 25% of the patients presented functional obstructive symptoms determined by the disease. The most frequent obstructive symptom was incomplete emptying of the bladder. Only the age of the patients and control group were correlated with urinary dysfunction. PMID- 12894268 TI - Primary headache diagnosis among chronic daily headache patients. AB - Chronic daily headache (CDH) refers to a group of non-paroxysmal daily or near daily headaches with peculiar characteristics that are highly prevalent in populations of neurological clinics and not uncommon among non-patient populations. Most of the patients with CDH had, as primary diagnosis, episodic migraine, which, with the time, presented a progressive frequency, pattern modification and loss of specific migraine characteristics. Other CDH patients had chronic tension-type headache, new daily persistent headache and hemicrania continua, which evolved thru the time to the daily or near-daily presentation. The objective of this study was to determine the primary headache diagnosis among a population of chronic daily headache patients attending a tertiary center for headache treatment. During a 5-year period 651 consecutive chronic daily headache patients attending a private subspecialty center were studied prospectively. The criteria adopted were those proposed by Silberstein et al (1994, revised 1996). Five hundred seventy four patients (88.1%) had episodic migraine as primary headache before turning into daily presentation, 52 (8%) had chronic tension-type headache, 14 (2.2%) had hemicrania continua and 11 patients (1.7%) had new daily persistent headache. CDH is quite frequent in patients from clinic-based studies suggesting a high degree of disability. Emphasis on education of patients suffering from frequent primary headaches with regard to measures that are able to decrease suffering and disability as well as better medical education directed to more efficient ways to handle these patients are necessary to improve outcome of such a prevalent condition. PMID- 12894269 TI - Basilar impression, Chiari malformation and syringomyelia: a retrospective study of 53 surgically treated patients. AB - The present study shows the results of 53 patients who have been treated surgically for basilar impression (BI), Chiari malformation (CM), and syringomyelia (SM). The patients were divided into two groups. Group I (24 patients) underwent osteodural decompression with large inferior occipital craniectomy, laminectomy from C 1 to C 3, dural opening in Y format, dissection of arachnoid adhesion between the cerebellar tonsils, medulla oblongata and spinal cord, large opening of the fourth ventricle and dural grafting with the use of bovine pericardium. Group II patients (29 patients) underwent osteodural neural decompression with the same procedures described above plus dissection of the arachnoid adherences of the vessels of the region of the cerebellar tonsils, and tonsillectomy (amputation) in 10 cases, and as for the remainning 19 cases, intrapial aspiration of the cerebellar tonsils was performed. The residual pial sac was sutured to the dura in craniolateral position. After completion of the suture of the dural grafting, a thread was run through the graft at the level of the created cisterna magna and fixed to the cervical aponeurosis so as to move the dural graft on a posterior- caudal direction, avoiding, in this way, its adherence to the cerebellum. PMID- 12894270 TI - Intraventricular pressure monitoring in patients with thalamic and ganglionic hemorrhages. AB - In the present study, we have evaluated the use of intraventricular pressure catheters in thalamic and ganglionic hemorrhages. Ten patients admitted in our Emergency Department in Glasgow Coma Scale (GCS) equal or below 13 enrolled the study (at least one point should have been lost in the eye opening score to exclude purely aphasic patients that were fully alert). After a complete clinical and neurological evaluation, computed tomography scans were obtained and the volume of the hematomas, as well as presence or absence of hydrocephalus, were considered. Intraventricular pressure catheters connected in parallel to external derivation systems were implanted and patients were thereafter sent to the ICU. Patients that presented mass effect lesions with sustained increased ICP levels or clinical and neurological deterioration were submitted in addition, to the surgical evacuation of the hematomas. Clinical evolution, complications and the rehabilitation of the patients were recorded. Clinical outcome was assessed with the Glasgow Outcome Score. In all but three patients the initial intracranial pressure levels were bellow 20 mmHg (mean for all patients was 14.1 +/- 6.5 mmHg). Notwithstanding, these three patients were extremely difficult to treat. For this group of patients mortality was 100%. Among the patients that presented ICP levels bellow 20 mmHg, 04 developed hydrocephalus and 03 did not display ventricular dilation. As expected, the major benefits concerning the intraventricular pressure catheters connected in parallel with external derivation systems were observed in the group of patients that presented ICP levels bellow 20 mmHg and had hydrocephalus. Mild non-statistically significant correlations for all the three groups were achieved either when the initial GCS and ICP levels (r=-0.28, p=0.43) or when ICP levels and the volumes of the hematomas were compared (r=0.38, p=0.28). In addition, no significant correlations were observed concerning the final outcome of the patients and the variables previously evaluated. PMID- 12894271 TI - [Neuroimaging abnormalities in parkinsonism: study of five cases]. AB - We report the brain magnetic resonance (MR) imaging abnormalities observed at the basal ganglia system of 5 patients (2 female and 3 male), who fulfilled the criteria of parkinsonism. The onset of parkinsonian syndrome ranged from 5 to 52 years old. All patients underwent MR exams with a 1.5T MR equipment. High field T2-weighted sequences disclosed hypersignal bilateral and symmetrically located exclusively at substantia nigra (3 cases), exclusively at globus pallidus (1case) and simultaneously at substantia nigra, globus pallidus and nigro-strital interconnections (1case). For three patients, the diagnose of secondary parkinsonism was supported by clinical data: the first had the onset of the symptoms after the exposure to an herbicide (glyphosate); the second after vaccination against measles; the third after coma due to encephalitis. For the other two patients, the onset of PS was progressive, resembling a typical idiopathic Parkinson's disease (PD) but the findings at the MR dimissed this initial diagnose. In this study, the contribution of neuroimaging was crucial to recognize secondary parkinsonism though the ethiological agents could not be determined in these patients. PMID- 12894272 TI - [Polysomnographic evaluation of clinical patients suffering from mood disturbance]. AB - The sleep patterns of sixty patients suffering from mood disturbance, were studied, with mean age of 36.1 +/- 11.3 years ( 12 males, 48 females). These patients were submitted to two nights of nocturnal polysomnographic evaluation; the second night acquired data was employed in the statistics. The following parameters were analyzed: parameters of sleep continuity, parameters of sleep architecture, REM and non-REM sleep parameters. The patients were analyzed in accordance to depression severity, presence of sleep disorders and relevant mood disturbances. The most common diagnosis was dysthymia in 25 cases (41.6%). Descriptive analysis was used to delineate the study. The statistics evaluation used the Fisher exact test and the Kruskal-Wallis statistics heterogeneity test. There was reduction in sleep continuity and increase in REM density, both not statistic significant. Stage 4 sleep showed significant reduction in severely depressed patients (p=0.0253). PMID- 12894273 TI - [Auditory evoked potentials evaluation of the brain stem in multiple sclerosis]. AB - The aim of this study was to evaluate the incidence of auditory and neurotological disorders in multiple sclerosis (MS). Twenty female and 9 male with a definite diagnosis of multiple sclerosis without signs of involvement of the brain stem underwent an audiological and an early auditory evoked potentials (EAEP). The wave forms were classified according to Jerger's (1986) classification. In 58 EAEP it was found 55% of type I (normal response) according to Jerger's classification in both sexes. Considering as an abnormal response the EAEP classified in type II, III, IV or V according to Jerger in at least one side, it was found 60% of abnormalities in females and 56% in males, totalizing 58,62% of all the studied subjects. The authors emphasize the use of EAEP in MS. PMID- 12894274 TI - [Clinical aspects of neurocysticercosis at semi-desert region of Brazilian northeast]. AB - Case sheets of 44 patients with neurocysticercosis, coming from Campina Grande and others cities of Para ba state, examined between 1990 - 2001 were analyzed. The average age (SD +/-) was 20.6 +/- 14.3 years old, of which 54.5% were male. Thirty eight patients (86.2%) came from urban area. The initial symptom was convulsion in 90.9% of the cases and headaches in 9.1%. The epileptical form was present in 63.6% of the cases and the combined form in 22.3%. Computerized tomography of the skull was compatible with neurocysticercosis in 100% of the cases, showing calcification in (59.1%), integral cysts in (18.2%) and in degenerating in (20.4%), isolated or associated. The LCR performed on 29 patients showed alterations in 25 (86.2%) cases, predominating linfomonocitary pleocitose in 100% of the cases and positive immunological reactions in 64.3% of the cases. In conclusion neurocysticercosis is a frequent cause for convulsions in children and young adults in this region, and the ocorrence in urbane and rural areas denote the poor control of the taeniases/cistycercosis complex in this state. PMID- 12894275 TI - [Classification of seizures in childhood obtained from seizures symptomatology descriptions of parents and guardians]. AB - The aim of this study was to determine the frequency of seizures descriptions from parents and guardians of epileptic children which are useful for clinical classification of epileptic seizures. The data were obtained from 112 children using recurrent and spontaneous epileptic seizures as the selection criteria. The study was realized in two parts. In the part I the following aspects were studied: age of onset and the preictal, ictal and postictal symptoms. In the part II the specific kinds of seizures were classified by motor semiology, as convulsive and nonconvulsive, and by using the International League Against Epilepsy (ILAE,1981) scheme. The results showed that 42.9% patients present epileptic auras and 36.6% present lateralizing signs. Concerning the semiologic events were motor in 95.5%, neurovegetative in 56.3%, psychic in 32.1%, and neurosensorials in 4.5%. Finally, the ILAE schema classified the seizures as partial in 59.9%, as generalized in 27.2%, and as not classified in 12.5%. All findings demonstrate a good level of clinical semiology information from parents and guardians that allow us to classify 87.5% of the seizures, and identify auras and lateralized signs. Theses results permit us to conclud that if a standard protocol is used, the descriptions from parents and guardian of epileptic children appears to be very reliable to clinical classification of epileptic seizure. PMID- 12894276 TI - [Comparison of functional performance among children with Down syndrome and children with age-appropriate development at 2 and 5 years of age]. AB - OBJECTIVE: To compare the functional performance of Down's syndrome (DS) children with normally developing children (ND), at 2 and 5 years of age. METHOD: Forty children were allocated into four groups (n=10): 1) children with DS with 2 years of age; 2) children with DS with 5 years of age; 3) normal children with 2 years of age; 4) normal children with 5 years of age. Children were evaluated with the functional test PEDI, which quantifies children's performance (skills and independence) in three domains: self-care, mobility and social function. Two-way ANOVA was used to compare group means and to test interaction effects age x pathology. Pre-planned contrast analyses were used to identify the significant bivariate comparisons. RESULTS: Main factors (age and pathology) were significant in the three domains of skills and independence performances. The interaction factor age x pathology was significant in self-care and mobility skills, as well as in children's independence in mobility and social function. Contrast analyses showed that at two years of age, normal children's performance is superior to DS's children in all three domains of functional skills and independence. However, at five years of age, significant group differences were only observed in the domains of self-care and social function skills, and independence. CONCLUSION: Results show the areas of performance where the delay presented by DS children was functionally manifested, at two and five years of age. Data indicate that the observed group differences were influenced by age, keeping themselves changeable across the development. PMID- 12894277 TI - [Compliance with chronic pain treatment: study of demographic, therapeutic and psychosocial variables]. AB - The aims of this study were to identify the prevalence of compliance with drug therapy in patients with chronic pain and analyze the relationships between compliance and characteristics of drug therapy and psychosocial factors (beliefs regarding pain, health locus of control and depression). Thirty patients were evaluated 5 times over a period of 6 months. Total compliance occurred in 43.3% to 56.7% of the patients. Partial compliance and non-compliance were high (40.0% 56.7%). The index of compliance did not vary over the six months. Low compliance related to occurrence of side effects and beliefs that the control of health depended on the patient, pain is a disability, that pain means the presence of physical injury and solicitous behavior of others is desirable when there is manifestation of pain. Knowing the factors involved in compliance enables us to test interventions that optimize it. PMID- 12894278 TI - [Arteriovenous malformation-glioma association: study of four cases]. AB - We reviewed the clinical presentation, imaging and histopathologic findings in 4 patients with the diagnosis of arteriovenous malformation associated with glioma that were operated on from 1991 to 2000 in our institution. Four patients (2 males; age between 15 and 52 years) presented with progressive headache with clinical evidence of intracranial hypertension (in 3) and partial seizures (in 1). CT scan showed a brain tumor without any detectable pathologic vessels. Histologic examination revealed astrocytic tumors associated with arteriovenous malformation. No patient presented the vascular component intermixed with the tumor. The arteriovenous-glioma association is rare and must be identified by a clear demarcation between the malformation and the tumor. PMID- 12894279 TI - [Acute alterations of neurotransmitters levels in striatum of young rat after pilocarpine-induced status epilepticus]. AB - High doses of the muscarinic cholinergic agonist, pilocarpine, result in behavioural changes, seizures and status epilepticus in rats. The purpose of the present work is to invetigate the striatal neurotransmissors level in young rats after status epilepticus induced by pilocarpine. Wistar rats were treated with a single dose of pilocarpine (400mg/Kg; s.c.). Controls received saline. Young animals were closed observed for behavioural changes during 1 and 24h. In these periods, the animals that developed status epilepticus and didn't survive this acute phase of seizures had the brains removed and striatal neurotransmissors level determined by HPLC. The concentration of dopamine, serotonine, dihydroxyphenylacetic acid, 5-hydroxyindolacetic acid was reduced and an increase in 4-hydroxy-3-methoxy-phenylacetic acid was observed. These results suggest that cholinergic activation can interage with dopaminergic and serotonergic systems in acute phase of the convulsive process in immature striatum. PMID- 12894280 TI - [Trigeminal radiofrequency rhizotomy for the treatment of trigeminal neuralgia: results and technical modification]. AB - The purpose of this study was to evaluate the efficacy of radiofrequency trigeminal rhizotomy in treating 135 patients harboring trigeminal neuralgia, and to introduce a technical modification to guide the puncture of the foramen ovale. A hundred and one (74.8%) patients were treated with a single surgical procedure whereas the 34 (25.2%) remaining patients required two procedures. Follow-up ranges from 6 months to 15 years. Pain relief in the immediate postoperative was achieved in 131 (97.0%) patients. After the initial procedure, recurrence happened in 33 (24.5%) patients. The complications included decrease corneal reflex (4.4%), masseter paresis (2.2%), painful dysesthesia (1.5%) and anesthesia dolorosa (0.7%). The radiofrequency trigeminal rizhotomy is a low risk, highly effective and minimally invasive procedure. The use of the computerized tomography guided fluoroscopy turns foramen ovale's puncture easier, fast and precise. PMID- 12894281 TI - [Neurovascular structures of the posterior surface of the petrous pyramid: correlation with the approaches of the cerebellopontine angle]. AB - A topographic study of posterior surface of the petrous pyramid was performed in 20 human cadaveri heads. The distances between the neurovascular structures were measured in the points where they contact the posterior surface of the petrous pyramid. The study also points out the relationship between the bone landmarks and the transverse and the superior petrous sinuses. The result of this study was correlated with the approaches to the cerebellopontine angle. PMID- 12894282 TI - Congenital dermoid inclusion cyst over the anterior fontanel: report of three cases. AB - Congenital dermoid inclusion cyst over the anterior fontanel (CDIC) is an uncommon cystic lesion located over the anterior fontanel. It is a benign and curative lesion and most of the time, can be diagnosed at birth. From 1994 to 2001, three patients were operated with this kind of lesion and after reviewing the literature we found 229 cases and only 6 cases described in Brazil. Our objective in this study is to report three more cases. PMID- 12894283 TI - [Occlusion of the common carotid artery treated with a subclavian-internal carotid artery bypass]. AB - We describe the case of a 62 years old woman with several transient isquemic attacks. The neuroradiological study demonstrated occlusion of the common carotid artery with revascularization of the internal carotid artery by anastomosis with the inferior thyroid artery and an aneurysm of internal carotid artery at the emergency of the anterior choroidal artery. The patient was treated with a bypass between the subclavian artery and the internal carotid artery with complete regression of the symptoms. PMID- 12894284 TI - Cauda equina hemangioblastoma: case report. AB - Hemangioblastomas of the spinal cord are rare lesions, and those located at the cauda equina are even rarer. Most commonly these tumors are present in patients with von Hippel-Lindau (VHL) syndrome. We describe here the case of a 48 years old woman with a pure radicular hemangioblastoma, not associated with VHL, presenting with radicular pain, diagnosed with magnetic ressonance imaging (MRI) and submitted to total resection with a very good outcome. To our knownledge, this is the second report to describe the MRI aspect of histologically proved hemangioblastoma of the cauda equina in a patient without clinical criteria for VHL. PMID- 12894285 TI - Cavernous carotid artery pseudo-aneurysm treated by stenting in acromegalic patient. AB - We report on a case of endovascular management of pseudoaneurysm of the cavernous segment of the internal carotid artery with covered stent reconstruction. A 36 years-old woman with a history of previous transsphenoidal approach for pituitary macroadenoma and false aneurysma formation was studied in a protocol that included balloon test occlusion and cerebral blood flow evaluation. An endovascular covered stent deployment in the area of the carotid laceration was performed with isolation of the aneurysm from the circulation and maintenance of the carotid flow. Helical angio-CT and cerebral digital subtraction angiography showed the carotid preservation without stenosis in the stented area. In conclusion, endovascular stent reconstruction for post-transsphenoidal carotid artery laceration and false aneurysm is demonstrated as useful technical adjunct in the management strategy and with the potential for carotid sacrifice morbidity avoidance. PMID- 12894286 TI - Malignant paraganglioma with vertebral metastasis: case report. AB - A paraganglioma is a rare tumor, composed of chromaffin cells, groups of cells associated to the autonomous system. When the tumor occurs in the adrenal gland, it is called pheochromocitoma. The malignant paraganglioma is a very rare presentation; it is diagnosed by local recurrence after total resection of the primary mass, or findings of distant metastases. We present a case report of a 29 year-old woman with cervico-brachial pain. In 1995 she underwent a carotid body tumor resection. Magnetic resonance imaging (MRI), plain X-rays and computerized tomography scan revealed multiple lesions in C5, T5 and T12. She underwent a surgical procedure to correct the cervical lesion. The histological and immunohistochemical assays revealed a malignant paraganglioma. She received adjuvant radiotherapy, showing clinical improvement after treatment, presenting no symptoms after one year. The therapeutic approach is based on the total resection of the tumor. The treatment of distant metastases can be made with adjuvant measures such as conventional radiotherapy, I 131-MIBG, or chemotherapy, especially in malignant pheochromocitomas. PMID- 12894287 TI - Glioblastoma multiforme of the pineal region: case report. AB - PURPOSE: pineal region tumors are uncommon, and comprise more frequently three categories: germ cell, parenchymal cell and glial tumors. Most pineal gliomas are low-grade astrocytomas. Glioblastoma multiforme, the most aggressive and common brain tumor, is extremely rare at this location with only few cases reported. CASE DESCRIPTION: a 29-year-old woman with a two month history of headache, nuchal pain, fever, nausea and seizures and physical examination showing nuchal rigidity, generalized hypotony, hypotrophy and hyper-reflexia, Babinski sign and left VI cranial par palsy. CT scan examination revealed a ill-defined hypodense lesion at the pineal region with heterogeneous contrast enhancement. MRI showed a lesion at the pineal region infiltrating the right thalamic region. The patient underwent a right craniotomy with partial resection of the mass. The histological examination of paraffin-embedded material defined the diagnosis of glioblastoma multiforme. Post-operative radiotherapy was indicated but the patient refused the treatment and died two months afterwards. CONCLUSION: in spite of its rarity at this location, glioblastoma multiforme should be considered in the differential diagnosis of aggressive lesions at the pineal region. PMID- 12894288 TI - Holmes tremor in association with bilateral hypertrophic olivary degeneration and palatal tremor: chronological considerations. Case report. AB - Hypertrophic olivary degeneration (HOD) is a rare type of neuronal degeneration involving the dento-rubro-olivary pathway and presents clinically as palatal tremor. We present a 48 year old male patient who developed Holmes' tremor and bilateral HOD five months after brainstem hemorrhage. The severe rest tremor was refractory to pharmacotherapy and botulinum toxin injections, but was markedly reduced after thalamotomy. Magnetic resonance imaging permitted visualization of HOD, which appeared as a characteristic high signal intensity in the inferior olivary nuclei on T2- and proton-density-weighted images. Enlargement of the inferior olivary nuclei was also noted. Palatal tremor was absent in that moment and appears about two months later. The delayed-onset between insult and tremor following structural lesions of the brain suggest that compensatory or secondary changes in nervous system function must contribute to tremor genesis. The literature and imaging findings of this uncommon condition are reviewed. PMID- 12894289 TI - Neonatal seizures: the overlap between diagnosis of metabolic disorders and structural abnormalities. Case report. AB - Inborn metabolic errors (IME) and cortical developmental malformations are uncommon etiologies of neonatal seizures, however they may represent treatable causes of refractory epilepsy and for this reason must be considered as possible etiological factors. This case report aims to demonstrate the importance of neuroimaging studies in one patient with neonatal seizures, even when there are clues pointing to a metabolic disorder. CASE REPORT: A previously healthy 14 day old child started presenting reiterated focal motor seizures (FMS) which evolved to status epilepticus. Exams showed high serum levels of ammonia and no other abnormalities. A metabolic investigation was conducted with normal results. During follow-up, the patient presented developmental delay and left side hemiparesia. Seizures remained controlled with anti-epileptic drugs for four months, followed by relapse with repetitive FMS on the left side. Temporary improvement was obtained with anti-epileptic drug adjustment. At the age of 6 months, during a new episode of status epilepticus, high ammonia levels were detected. Other metabolic exams remained normal. The child was referred to a video-electroencephalographic monitoring and continuous epileptiform discharges were recorded over the right parasagittal and midline regions, with predominance over the posterior quadrant. A new neuroimaging study was performed and displayed a malformation of cortical development. Our case illustrates that because newborns are prone to present metabolic disarrangement, an unbalance such as hyperammonemia may be a consequence of acute events and conduct to a misdiagnosis of IME. PMID- 12894290 TI - [Growth hormone deficiency after treatment of medulloblastoma with radiotherapy in childhood: case report]. AB - Craniospinal radiation therapy for treatment of brain tumors may result in growth hormone (GH) insufficiency with resultant linear growth retardation, one of the most common complications. We report the case of a 10-year-old boy presenting headache associated to vertigo, nausea and vomiting. A CT scan showed an homogeneous mass in the left cerebelar hemisphere, that was surgically removed. The histopathological examination revealed medulloblastoma and the patient was submitted to craniospinal radiation. He did not present tumor recurrence nor neurological or cognitive deficits during 4 years, but evolved to short stature due to GH deficiency. Nowadays, he is being receiving GH 0,1 U/kg/day and has presented 4cm stature increment after 6 months. The present case highlights the importance in carefully monitoring of children after cranial radiation for oncologic treatment, because they may develop hormonal deficiencies, that can be successfully replaced. PMID- 12894291 TI - [Complication of ventriculoperitoneal shunting: inguinal hernia with scrotal migration of catheter. Case report]. AB - Ventriculoperitoneal shunting is the treatment of choice for hydrocephalus in its different etiologies. Mechanical failure and infections are common complications of shunting. The development of inguinal hernia or hydrocele after shunting is an uncommon condition and the migration of abdominal catheter into the scrotum rare. The patency of processus vaginalis, the raised intra-abdominal pressure and the age of patients are factors related with the genesis of these pathologies. This paper reports a case of a child who developed an inguinal hernia after ventriculoperitoneal shunting and scrotal migration of shunt. The literature is reviewed and treatment is discussed. PMID- 12894292 TI - [Foreign body in the orbital cone: case report]. AB - Orbital injuries may be associated with the presence of foreign bodies in which long retention leads to damages such as loss of vision or cerebral abscess. We present the case of a 21-year-old male patient that developed ptosis, limitation of movements and an inflammatory reaction in the right eye after being hit by a tree branch. A CT scan of the orbits revealed an increased density in the right orbital cone. After the neurosurgical approach for the removal of the foreign body, a wooden fragment measuring 2.0x0.3 cm, the patient had a good outcome without visual deficits, ptosis regression and resolution of the inflammatory process. The postoperative CT scan didn't identify any remaining foreign body fragment. This case calls attention to the fact that the wooden splinter presented on CT scan with high density (136 HU) two months after the trauma and was removed by a supraorbital craniotomy. PMID- 12894293 TI - [Hopkins' syndrome in the differential diagnosis of flaccid paralysis in children: clinical and neurophysiological features. Case report]. AB - INTRODUCTION: Hopkins syndrome is a motor neuron disease which leads to a flaccid paralysis affecting one or more limbs resembling poliomyelites. It follows an asthmatic attack and the prognosis is poor. All the 34 related cases occured after 13 months of age and there is no report in South America. Our objective is to describe a case of Hopkins Syndrome in Brazil affecting a patient younger than 1 year. CASE: Male 4 months-old infant, started presenting wheezing that turned into respiratory failure which required mecanical ventilation. Three days later he initiated with loss of strength in the lower limbs (LL). The patient was dismissed from hospital better of his respiratory complains but still paraparetic. Five months later, the patient returned with another episode of severe bronchospasm. At that time, he presented with flaccid paralysis, arreflexia and atrophy of the LL. There were no upper limbs and sensory abnormalities. The patient undergone to lumbar puncture, spinal MRI and SEPs which were all normal. MUSCLE BIOPSY: type grouping. EMG and NCS were in keeping with motor neuron disease affecting just the lumbosacral region. CONCLUSION: Hopkins syndrome should be included in the differential diagnosis of any flaccid paralysis when it is associated with an asthma attack. PMID- 12894294 TI - Cervical epidural haematoma with clivus fracture: case report. AB - Clivus fractures are rare and severe entities, usually associated with vascular or cranial nerve lesions and frequently diagnosed postmortem. Cervical epidural haematomas can be traumatic or spontaneous, manifested in acute or chronic form, and are treated surgically in the majority of cases, although the conservative treatment also can be indicated to patients with incomplete and non-progressive deficits. The authors report the case of a female patient, 8 years old, victim of trampling in public way by a high velocity motorized vehicle, admitted in Glasgow 7, anisocoric pupils (left pupil midriatic), whose radiological investigation showed a transverse fracture of the clivus, cervical epidural haematoma and diffuse axonal injury. The patient was submitted to intracranial pressure monitorization, sedation and conservative treatment with dexamethasone, with good outcome. The authors also present a literature review. PMID- 12894295 TI - When epilepsy may have changed history: Antonio Moreira Cesar as the commander of the third expedition in the war of Canudos. AB - Colonel Antonio Moreira Cesar, the Commander of the third Expedition against Canudos (1896-1897), nicknamed "head-chopper", was considered an implacable military man, a synonym of ferocity and extreme brutality against his adversaries. Therefore, he was nominated the Commander of an expedition considered almost invincible. Since his 30's he presented epileptic seizures, which increased in frequency and severity on his way to Canudos. After several well-documented episodes and probably considering himself the winner in anticipation, he ordered a premature and almost ingenuous attack against Canudos. His misjudging is attributed to the effect of successive seizures. He was shot and killed on the very first day of that battle and his expedition had a horrible and unexpected end. Based on the descriptions of his biographer we discuss the nature of his disease probably characterized by focal seizures with elementary and complex visual hallucinations followed by language deficits and episodes of complex partial seizures and secondary generalization and its role in this episode of Brazilian history. PMID- 12894300 TI - Evaluation of the catalytic activity of lipases immobilized on chrysotile for esterification. AB - In the present work, the ester synthesis in organic media catalyzed by lipases immobilized on chrysotile was studied. Lipases of different sources (Mucor javanicus, Pseudomonas cepacia, Rhizopus oryzae, Aspergillus niger and Candida rugosa) were immobilized on chrysotile, an inexpensive magnesium silicate, and used for esterification of hexanoic, octanoic and lauric acid with methanol, ethanol, 1-butanol and 1-octanol at 25 C in hexane as solvent. The best results were obtained with Mucor javanicus lipase and lauric acid giving yields of 62-97% of ester. PMID- 12894301 TI - Photoelectrochemical solar cell using extract of Eugenia jambolana Lam as a natural sensitizer. AB - The extract of Jambol o (java plum), Eugenia jambolana Lam, was used as a natural sensitizer of a wide band-gap semiconductor (TiO2) in photoelectrochemical solar cells. The natural dye, adsorbed onto the semiconductor surface, absorbs visible light and promotes electron transfer across the dye/semiconductor interface. Photogenerated current and voltage as high as 2.3 mA and 711 mV, respectively, were obtained and effective conversion of visible light into electricity was achieved. The use of a natural product as the semiconductor sensitizer enables a faster and simpler production of cheaper and environmentally friendly solar cells. PMID- 12894302 TI - Balanced cytokine-producing pattern in mice immunized with an avirulent Trypanosoma cruzi. AB - We have previously demonstrated that inoculation of BALB/c mice with trypomastigotes of CL-14, an avirulent Trypanosoma cruzi clone, prevents the development of parasitemia and mortality after challenge with virulent CL strain. In this report, we investigated the cytokine and antibody profiles induced by inoculation with CL-14 clone. Groups of mice were inoculated with trypomastigotes of CL-14 clone and challenged with infective CL strain. Challenged CL-14 inoculated mice had lower levels of IFN-gamma and higher production of IgG1 antibodies as compared to CL strain-infected mice. Previous inoculation with CL 14 clone partially prevented the suppression of IL-2 production caused by CL strain infection. No significant differences were found regarding IL-4 production by splenocytes from CL-14-inoculated or control groups after challenge with CL strain. Our results show that protection against acute T. cruzi infection induced by CL-14 inoculation correlates with a balanced T1/T2 cytokine production, a profile likely to be beneficial for the host. PMID- 12894308 TI - Characterization of newborns with nonimmune hydrops fetalis admitted to a neonatal intensive care unit. AB - PURPOSE: To determine the incidence and characteristics of nonimmune hydrops fetalis in the newborn population. METHOD: A retrospective study of the period between 1996 and 2000, including all newborns with a prenatal or early neonatal diagnosis of nonimmune hydrops fetalis, based on clinical history, physical examination, and laboratory evaluation. The following were analyzed: prenatal follow-up, delivery type, gender, birth weight, gestational age, presence of perinatal asphyxia, nutritional classification, etiopathic diagnosis, length of hospital stay, mortality, and age at death. RESULTS: A total of 47 newborns with hydrops fetalis (0.42% of live births), 18 (38.3%) with the immune form and 29 (61.7%) with the nonimmune form, were selected for study. The incidence of nonimmune hydrops fetalis was 1 per 414 neonates. Data was obtained from 21 newborns, with the following characteristics: 19 (90.5%) were suspected from prenatal diagnosis, 18 (85.7%) were born by cesarean delivery, 15 (71.4%) were female, and 10 (47.6%) were asphyxiated. The average weight was 2665.9 g, and the average gestational age was 35 3/7 weeks; 14 (66.6%) were preterm; 18 (85.0 %) appropriate delivery time; and 3 (14.3%) were large for gestational age. The etiopathic diagnosis was determined for 62%, which included cardiovascular (19.0%), infectious (9.5%), placental (4.8%), hematologic (4.7%), genitourinary (4.8%), and tumoral causes (4.8%), and there was a combination of causes in 9.5%. The etiology was classified as idiopathic in 38%. The length of hospital stay was 26.6 +/- 23.6 days, and the mortality rate was 52.4%. CONCLUSIONS: The establishment of a suitable etiopathic diagnosis associated with prenatal detection of nonimmune hydrops fetalis can be an important step in reducing the neonatal mortality rate from this condition. PMID- 12894309 TI - Conventional approach x laparoscopic abdominoperineal resection for rectal cancer treatment after neoadjuvant chemoradiation: results of a prospective randomized trial. AB - OBJECTIVE: The aims of this study were to evaluate the safety and efficacy of laparoscopic abdominoperineal resection compared to conventional approach for surgical treatment of patients with distal rectal cancer presenting with incomplete response after chemoradiation. METHOD: Twenty eight patients with distal rectal adenocarcinoma were randomized to undergo surgical treatment by laparoscopic abdominoperineal resection or conventional approach and evaluated prospectively. Thirteen underwent laparoscopic abdominoperineal resection and 15 conventional approach. RESULTS: There was no significant difference (p<0,05) between the two studied groups regarding: gender, age, body mass index, patients with previous abdominal surgeries, intra and post operative complications, need for blood transfusion, hospital stay after surgery, length of resected segment and pathological staging. Mean operation time was 228 minutes for the laparoscopic abdominoperineal resection versus 284 minutes for the conventional approach (p=0.04). Mean anesthesia duration was shorter (p=0.03) for laparoscopic abdominoperineal resection when compared to conventional approach : 304 and 362 minutes, respectively. There was no need for conversion to open approach in this series. After a mean follow-up of 47.2 months and with the exclusion of two patients in the conventional abdominoperineal resection who presented with unsuspected synchronic metastasis during surgery, local recurrence was observed in two patients in the conventional group and in none in the laparoscopic group. CONCLUSIONS: We conclude that laparoscopic abdominoperineal resection is feasible, similar to conventional approach concerning surgery duration, intra operative morbidity, blood requirements and post operative morbidity. Larger number of cases and an extended follow-up are required to adequate evaluation of oncological results for patients undergoing laparoscopic abdominoperineal resection after chemoradiation for radical treatment of distal rectal cancer. PMID- 12894310 TI - Use of tacrolimus in rescue therapy of acute and chronic rejection in liver transplantation. AB - PURPOSE: To study the indications and results of tacrolimus as rescue therapy for acute cellular or chronic rejection in liver transplantation. PATIENTS AND METHODS: Eighteen liver transplant recipients who underwent rescue therapy with tacrolimus between March 1995 and August 1999 were retrospectively studied. The treatment indication, patients, and graft situation were recorded as of October 31st, 1999. The response to tacrolimus was defined as patient survival with a functional graft and histological reversal of acute cellular, or for chronic rejection, bilirubin serum levels decreasing to up to twice the upper normal limit. RESULTS: Fourteen cases (77.8%) presented a good response. The response rate for the different indications was: (1) acute cellular + sepsis - 0/1 case; (2) recurrent acute cellular - 1/1 case; (3) OKT3-resistant acute cellular - 2/2 cases; (4) steroid-resistant acute cellular + active viral infection - 3/3 cases; (5) chronic rejection - 8/11 cases (72.7% response rate). The 4 patients who did not respond died. CONCLUSION: Tacrolimus rescue therapy was successful in most cases of acute cellular and chronic rejection in liver transplantation. PMID- 12894311 TI - Histopathologic and biochemical liver test abnormalities in chronic asymptomatic or oligosymptomatic alcoholics: a review. AB - PURPOSE: To review the medical literature regarding the histopathologic and biochemical liver test abnormalities in chronic asymptomatic or oligosymptomatic alcoholics. METHODS: Review of articles in the MEDLINE and LILACS databases regarding serum levels and prevalence of alterations in aspartate aminotransferase, alanine-aminotransferase, alkaline phosphatase, and total bilirubin, in relation to liver histopathology, with or without discrimination of types of histopathologic alteration. RESULTS: Global mean prevalence rates of aspartate-aminotransferase and alanine-aminotransferase alterations were 86.3% and 51.1%; in cases with steatosis they were 79.1% and 38.5%; and in cases of hepatitis, 90.1% and 58%. In all studies, prevalence rates of aspartate aminotransferase alterations were significantly higher with lower variability than those of alanine-aminotransferase. Mean aspartate-aminotransferase levels were higher than 2N (N is the upper normal limit of the method employed) in all cases with hepatitis histopathology, while those of alanine-aminotransferase were 1.48N, in the same cases. Prevalence of alkaline phosphatase and total bilirubin abnormalities were 74.5% and 74.9% globally; in cases of steatosis, they were 70.9% and 67.9%; and in cases of hepatitis, 75.9% and 77.7%. Mean alkaline phosphatase levels were above the upper normal limit in all cases, but those of total bilirubin were above normal in 4 of 7 hepatitis studies. CONCLUSIONS: Prevalence of aspartate-aminotransferase alteration was consistently related to presence of histopathologic abnormalities; an enzyme level higher than 2N suggests the diagnosis of alcoholic hepatitis. PMID- 12894312 TI - Antiphospholipid antibodies in 57 children and adolescents with systemic lupus erythematosus. AB - OBJECTIVE: To investigate the frequencies and behavior of antiphospholipid antibodies in 57 children and adolescents with systemic lupus erythematosus. METHODS: Anticardiolipin antibodies were investigated by ELISA and lupus anticoagulant antibodies by the international tests recommended. The antiphospholipid antibodies analyses were performed in frozen samples (mean of 5.3 samples per patient obtained during a mean follow-up period of 3 years and 7 months) and on blood samples collected between January 1997 and November 1998 (mean of 2.5 samples per patient during a 2-year follow-up period). RESULTS: The frequencies of antiphospholipid antibodies (anticardiolipin and lupus anticoagulant) were similar in the samples collected prospectively and in the frozen samples (retrospective study): 63.2% and 75.4% respectively. Positivity for these antibodies fluctuated during the follow-up period and was not associated with any clinical or laboratory parameters of lupus erythematosus, including autoantibodies and also including disease activity and/or severity scores. CONCLUSIONS: The frequencies of antiphospholipid antibodies in children and adolescents with lupus erythematosus were similar to those observed in adults. The positivity fluctuated during the follow-up and was not correlated with clinical and/or laboratory disease parameters. PMID- 12894313 TI - Dura mater mitral and tricuspid bioprostheses: 30 years of follow-up. AB - PURPOSE: The dura mater bioprosthesis was developed in the Department of Cardiopneumology of the Hospital das Clinicas of the University of Sao Paulo Medical School in 1971. Here, we present the clinical results of the dura mater bioprosthesis over 30 years of follow-up. METHODS: We studied 70 consecutive patients who underwent mitral or tricuspid valve replacement with a dura mater bioprosthesis between January 1971 and August 1972. RESULTS: The early mortality was 10% (7 patients). The follow-up was 87% complete (9 patients were lost to follow-up). Two patients were alive and asymptomatic 30 years after valve replacement; 33 patients underwent reoperations due to valve dysfunction, and 19 died during the follow-up period. At 30 years, the actuarial survival was 49.2 +/ 8.6%; freedom from rupture, 27.0 +/- 10.2%; freedom from calcification, 78.8 +/- 8.6%; and freedom from reoperation, 18.8 +/- 7.5%. CONCLUSIONS: The dura mater bioprosthesis played an important role in the treatment of patients with mitral and tricuspid valve disease. The low rate of thromboembolism and the long period of follow-up without evidence of valve dysfunction, which occurred for several of our patients, are important characteristics of these bioprosthesis. PMID- 12894314 TI - Meningeal carcinomatosis as the initial manifestation of a gallbladder adenocarcinoma associated with a Krukenberg tumor. AB - A case of malignant neoplasm is described in which the initial manifestations were mental dysfunction and meningeal irritation, mimicking chronic or subacute meningitis. Physical examination showed cranial nerve involvement and a pelvic tumor. There was progressive deterioration, and death occurred in 2 weeks. The autopsy revealed a gallbladder adenocarcinoma, meningeal carcinomatosis, and ovarian metastasis presenting as a Krukenberg tumor. The authors emphasize the importance of including meningeal carcinomatosis as a possibility in the differential diagnosis of non-characteristic clinical pictures, as well as the importance of the cerebrospinal fluid cytologic examination, repeated as needed, in order to confirm this diagnosis. PMID- 12894315 TI - Measuring male infertility: epidemiological aspects. AB - Evidence suggests that human semen quality may have been deteriorating in recent years. Most of the evidence is retrospective, based on analysis of data sets collected for other purposes. Measures of male infertility are needed if we want to monitor the biological capacity for males to reproduce over time or between different populations. We also need these measures in analytical epidemiology if we want to identify risk indicators, risk factors, or even causes of an impaired male fecundity-that is, the male component in the biological ability to reproduce. The most direct evaluation of fecundity is to measure the time it takes to conceive. Since the time of conception may be missed in the case of an early abortion, time to get pregnant is often measured as the time it takes to obtain a conception that survives until a clinically recognized pregnancy or even a pregnancy that ends with a live born child occurs. A prolonged time required to produce pregnancy may therefore be due to a failure to conceive or a failure to maintain a pregnancy until clinical recognition. Studies that focus on quantitative changes in fecundity (that does not cause sterility) should in principle be possible in a pregnancy sample. The most important limitation in fertility studies is that the design requires equal persistency in trying to become pregnant and rather similar fertility desires and family planning methods in the groups to be compared. This design is probably achievable in exposure studies that make comparisons with reasonable comparable groups concerning social conditions and use of contraceptive methods. PMID- 12894316 TI - Critical analysis of the allocation policy for liver transplantation in Brazil. AB - Liver transplantation is now the standard treatment for end-stage liver disease. Given the shortage of liver donors and the progressively higher number of patients waiting for transplantation, improvements in patient selection and optimization of timing for transplantation are needed. Several solutions have been suggested, including increasing the donor pool; a fair policy for allocation, not permitting variables such as age, gender, and race, or third party payer status to play any role; and knowledge of the natural history of each liver disease for which transplantation is offered. To observe ethical rules and distributive justice (guarantee to every citizen the same opportunity to get an organ), the "sickest first" policy must be used. Studies have demonstrated that death has no relationship with waiting time, but rather with the severity of liver disease at the time of inclusion. Thus, waiting time is no longer part of the United Network for Organ Sharing distribution criteria. Waiting time only differentiates between equally severely diseased patients. The authors have analyzed the waiting list mortality and 1-year survival for patients of the State of S o Paulo, from July 1997 through January 2001. Only the chronological criterion was used. According to "Secretaria de Estado da Saude de Sao Paulo" data, among all waiting list deaths, 82.2% occurred within the first year, and 37.6% within the first 3 months following inclusion. The allocation of livers based on waiting time is neither fair nor ethical, impairs distributive justice and human rights, and does not occur in any other part of the world. PMID- 12894317 TI - Perspectives on health impact assessment. PMID- 12894318 TI - Community participation in health impact assessments: intuitively appealing but practically difficult. PMID- 12894319 TI - Health impact assessment--how to start the process and make it last. PMID- 12894320 TI - Health impact assessment: the wider context. PMID- 12894321 TI - Health impact assessment of agriculture and food policies: lessons learnt from the Republic of Slovenia. AB - The most important public health priority in agricultural policy-making is currently food safety, despite the relatively higher importance of food security, nutrition, and other agricultural-related health issues in terms of global burden of disease. There is limited experience worldwide of using health impact assessment (HIA) during the development of agriculture and food policies, which perhaps reflects the complex nature of this policy sector. This paper presents methods of HIA used in the Republic of Slovenia, which is conducting a HIA of proposed agricultural and food policies due to its accession to the European Union. It is the first time that any government has attempted to assess the health effects of agricultural policy at a national level. The HIA has basically followed a six-stage process: policy analysis; rapid appraisal workshops with stakeholders from a range of backgrounds; review of research evidence relevant to the agricultural policy; analysis of Slovenian data for key health-related indicators; a report on the findings to a key cross-government group; and evaluation. The experience in Slovenia shows that the HIA process has been a useful mechanism for raising broader public health issues on the agricultural policy agenda, and it has already had positive results for policy formation. HIA is one useful approach to more integrated policy-making across sectors, but clearly it is not the only mechanism to achieve this. A comparison of the approach used in Slovenia with HIA methods in other countries and policy contexts shows that there are still many limitations with HIA application at a government level. Lessons can be learnt from these case studies for future development and application of HIA that is more relevant to policy-makers, and assists them in making more healthy policy choices. PMID- 12894322 TI - Including health in transport policy agendas: the role of health impact assessment analyses and procedures in the European experience. AB - From the mid-1990s, research began to highlight the importance of a wide range of health impacts of transport policy decisions. The Third Ministerial Conference on Environment and Health adopted a Charter on Transport, Environment and Health based on four main components: bringing awareness of the nature, magnitude and costs of the health impacts of transport into intergovernmental processes; strengthening the arguments for integration of health into transport policies by developing in-depth analysis of the evidence; developing national case studies; and engaging ministries of environment, health and transport as well as intergovernmental and nongovernmental organizations. Negotiation of the Charter was based on two converging processes: the political process involved the interaction of stakeholders in transport, health and environment in Europe, which helped to frame the issues and the approaches to respond to them; the scientific process involved an international group of experts who produced state-of- the-art reviews of the health impacts resulting from transportation activities, identifying gaps in existing knowledge and methodological tools, specifying the policy implications of their findings, and suggesting possible targets for health improvements. Health arguments were used to strengthen environmental ones, clarify costs and benefits, and raise issues of health equity. The European experience shows that HIA can fulfil the need for simple procedures to be systematically applied to decisions regarding transport strategies at national, regional and local levels. Gaps were identified concerning models for quantifying health impacts and capacity building on how to use such tools. PMID- 12894323 TI - Soft data, hard effects. Strategies for effective policy on health impact assessment--an example from the Netherlands. AB - Health impact assessment (HIA) on a strategic level focuses on the broad determinants of health. However, the evidence with regard to the health impacts is often necessarily 'soft'. The example of a health impact review on national housing policy in the Netherlands shows that HIA can be effective even in the absence of hard data. Strategies used to overcome the problem of not having hard data are outlined. The authors argue that, for HIA to be effective, it does not necessarily have to be limited to easy-to-measure, easy-to-quantify programmes and health effects. PMID- 12894324 TI - Towards health impact assessment of drinking-water privatization--the example of waterborne carcinogens in North Rhine-Westphalia (Germany). AB - Worldwide there is a tendency towards deregulation in many policy sectors - this, for example, includes liberalization and privatization of drinking-water management. However, concerns about the negative impacts this might have on human health call for prospective health impact assessment (HIA) on the management of drinking-water. On the basis of an established generic 10-step HIA procedure and on risk assessment methodology, this paper aims to produce quantitative estimates concerning health effects from increased exposure to carcinogens in drinking water. Using data from North Rhine-Westphalia in Germany, probabilistic estimates of excess lifetime cancer risk, as well as estimates of additional cases of cancer from increased carcinogen exposure levels are presented. The results show how exposure to contaminants that are strictly within current limits could increase cancer risks and case-loads substantially. On the basis of the current analysis, we suggest that with uniform increases in pollutant levels, a single chemical (arsenic) is responsible for a large fraction of expected additional risk. The study also illustrates the uncertainty involved in predicting the health impacts of changes in water quality. Future analysis should include additional carcinogens, non-cancer risks including those due to microbial contamination, and the impacts of system failures and of illegal action, which may be increasingly likely to occur under changed management arrangements. If, in spite of concerns, water is privatized, it is particularly important to provide adequate surveillance of water quality. PMID- 12894325 TI - Evaluation as a key part of health impact assessment: the English experience. AB - Health impact assessment (HIA) is an approach that has experienced increased activity and wide support within England. This is reflected in the large number of HIAs being undertaken at local and regional levels, by the advocacy of HIA in many national policy documents, and by the setting up of specialized HIA posts and centres. To continue this level of support, and to justify any increase, the approach of HIA must show whether and how the HIA approach informs the decision making process and, in particular, whether it improves health and reduces health inequalities. The first steps in answering these questions have been taken by the partial evaluation of some completed HIAs within England. A description of these evaluations is presented. Work is also progressing on promising practice guidance for practitioners on the topic of evaluating HIAs. The further development of evaluation within HIA is required, at both a practitioner and research level, and provides an exciting opportunity for people and groups to advance the evidence base for HIA, while also proving its place as an effective public health intervention. PMID- 12894326 TI - Health and equity impacts of a large oil project in Africa. AB - A system of external reviewers was established by the World Bank Group to promote a thorough environmental and health impact assessment for the 3.5 billion US dollars Chad Oil Export Project, based on a loan request from Chad, Cameroon and a consortium of oil companies. The environmental and health assessment process showed evidence of its ability to minimize the number of deaths from malaria, traffic accidents and construction accidents and the occurrence of minor sexually transmitted diseases, diarrhoeal diseases and respiratory diseases; it also probably limited adverse impacts on wildlife and tropical ecology along the pipeline route. However, the system was unable to deal with the larger issues, which included: the intrinsic unsustainability of this kind of extraction project; its eventual contribution to large amounts of greenhouse gases in the atmosphere; the lack of equity in sharing the risks, negative impacts, benefits and decision-making among the various participants in the project; and the possible acceleration of transmission of the AIDS virus into central Africa. Unfortunately, the international panel of experts appointed by the World Bank Group was largely ignored by the project proponents, and had little success in minimizing the most serious impacts or in improving the social equity of the project. PMID- 12894327 TI - Possible illnesses: assessing the health impacts of the Chad Pipeline Project. AB - Health impact assessments associated with large-scale infrastructure projects, such as the Chad-Cameroon Petroleum Development and Pipeline Project, monitor pre existing conditions and new diseases associated with particular industries or changes in social organization. This paper suggests that illness self-reports constitute a complementary set of benchmarks to measure the health impacts of these projects, and presents data gathered in ongoing household and health service surveys in Ngalaba, a village near a major oilfield in Chad. In an initial 16-week period of weekly data collection, 363 people reported few of the clinically chronic or asymptomatic conditions expected according to health transition theory, and the overall level of illness reporting was low. Illnesses often were described by symptoms or lay diagnoses. Health care practitioners were consulted rarely; when they were, resources for diagnosis and treatment were limited. Clinically acute, short-duration illnesses (e.g. parasitic infections, toothaches, or hernias) were experienced as chronic conditions and were reported week after week. The low levels of illness reporting and lack of clinically chronic conditions are not taken to mean that rural Chadians are healthy. Rather, the patterns of morbidity reflect a particular local ecology in which health services are organized and care dispensed in ways that limit the possibilities for illness in terms of types of illnesses that can be diagnosed and reported, forms illnesses take, and ways in which illnesses are experienced. Illness self reports are useful adjuncts to "harder" biological measures in HIAs, particularly in the context of large-scale infrastructure projects with explicit development goals. Rather than providing data on the extent to which harm has been mitigated by corporate, state, and donor activities, self-reports show the possibilities of illness in local contexts. PMID- 12894328 TI - Integrated environmental impact assessment: a Canadian example. AB - The Canadian federal process for environmental impact assessment (EIA) integrates health, social, and environmental aspects into either a screening, comprehensive study, or a review by a public panel, depending on the expected severity of potential adverse environmental effects. In this example, a Public Review Panel considered a proposed diamond mining project in Canada's northern territories, where 50% of the population are Aboriginals. The Panel specifically instructed the project proposer to determine how to incorporate traditional knowledge into the gathering of baseline information, preparing impact prediction, and planning mitigation and monitoring. Traditional knowledge is defined as the knowledge, innovations and practices of indigenous and/or local communities developed from experience gained over the centuries and adapted to local culture and environment. The mining company was asked to consider in its EIA: health, demographics, social and cultural patterns; services and infrastructure; local, regional and territorial economy; land and resource use; employment, education and training; government; and other matters. Cooperative efforts between government, industry and the community led to a project that coordinated the concerns of all interested stakeholders and the needs of present and future generations, thereby meeting the goals of sustainable development. The mitigation measures that were implemented take into account: income and social status, social support networks, education, employment and working conditions, physical environments, personal health practices and coping skills, and health services. PMID- 12894329 TI - Health impact assessment needs in south-east Asian countries. AB - A situation analysis was undertaken to assess impediments to health impact assessment (HIA) in the South-East Asia Region of WHO (SEARO). The countries of the region were assessed on the policy framework and procedures for HIA, existing infrastructure required to support HIA, the capacity for undertaking HIA, and the potential for intersectoral collaboration. The findings show that environmental impact assessment (EIA) is being used implicitly as a substitute for HIA, which is not explicitly or routinely conducted in virtually all countries of the Region. Therefore, policy, infrastructure, capacity, and intersectoral collaboration need strengthening for the routine implementation of HIA. PMID- 12894330 TI - Transport and land-use policies in Delhi. AB - Current transportation policies in mega-cities worldwide lead to major threats to health through traffic injuries, air pollution, noise, reduction in physical activities, and adverse impact on urban quality of life. In addition, a large section of the population in cities in low-income countries has to live in informal-sector, substandard housing. Many transportation policies fail to take enough account of their impacts on poverty and social exclusion, and they neglect the access and transportation demands of the more economically disadvantaged groups of society, who rely mostly on public transportation, walking, and cycling. Delhi, the capital city of India, is an interesting case because failure to consider the broad spectrum of health effects that may result from transport and land-use policies and investments has resulted in decisions that penalize the least affluent groups of the population and make it more difficult for them to get to jobs, education, health care, amenities, and services. PMID- 12894331 TI - Comparative assessment of transport risks--how it can contribute to health impact assessment of transport policies. AB - Health impact assessment (HIA) and comparative risk assessment (CRA) are important tools with which governments and communities can compare and integrate different sources of information about various health impacts into a single framework for policy-makers and planners. Both tools have strengths that may be combined usefully when conducting comprehensive assessments of decisions that affect complex health issues, such as the health risks and impacts of transport policy and planning activities. As yet, however, HIA and CRA have not been applied widely to the area of transport. We draw on the limited experience of the application of these tools in the context of road transport to explore how comparative assessment of transport risks can contribute to HIA of transport policies. PMID- 12894332 TI - Complexity and rigour in assessing the health dimensions of sectoral policies and programmes. PMID- 12894333 TI - What can health impact assessment add to comparative risk assessment in decision making? PMID- 12894334 TI - Health impact assessment in international development assistance: the World Bank experience. PMID- 12894335 TI - Developing health impact assessment in the European Union. PMID- 12894336 TI - Development of health impact assessment in Thailand: recent experiences and challenges. PMID- 12894337 TI - Trends in lead content of petrol in Pakistan. PMID- 12894338 TI - Nutritionists unimpressed by sugar lobby's outcry. PMID- 12894339 TI - GlaxoSmithKline, under pressure, cuts price of AIDS treatment for poor countries. PMID- 12894341 TI - Farmers kill 23 million birds to stop influenza virus. PMID- 12894346 TI - Distinct genetic and epigenetic changes in medullary breast cancer. AB - Genetic instability resulting in chromosome aneuploidy or mismatch repair deficiency characterizes cancer. Medullary carcinoma (MC) of the breast is a specific form of breast cancer with unique clinical, epidemiologic, and prognostic features, suggesting distinctive tumorigenic pathways. To investigate the nature of the genetic changes associated with MC we analyzed a series of 22 tumors. Chromosomal imbalances were assessed by comparative genomic hybridization (CGH) and mismatch repair (MMR) deficiency tested for through assessment of microsatellite instability (MSI) and expression of MLH1 and MSH2 genes. MMR deficiency was detected in only a small proportion of cases. The chromosomal copy number changes showed some similarities to BRCA1-associated tumors. A high level of BRCA1 promoter hypermethylation was detected, suggesting a possible role of this gene in MC development. PMID- 12894347 TI - Overexpression of NSAID-activated gene product in prostate cancer. AB - NSAID-activated gene (NAG-1) protein was previously identified by microarray analysis as overexpressed in prostate cancer. We performed immunohistochemistry and Western blotting with rabbit polyclonal antibody to NAG-1. Fifty malignant tissues obtained by prostatectomy and 17 from benign cases were compiled. Cancer tissues included Gleason scores 3-6, 3+4=7, 4+3=7, and 8-10. Cancer and high grade prostatic intraepithelial neoplasia (PIN) consistently showed moderate to intense cytoplasmic reactivity in 95-100% of epithelium. Staining intensity inversely correlated with preoperative serum prostate-specific antigen (PSA) (p=0.005) and with grade, averaging (on a 0 to 3+ scale) 2.3 +/- 0.6 in the lowest grade group, and 2.0 +/- 0.7, 1.8 +/- 0.5, and 1.5 +/- 0.6 as grade increased (p<0.008). Benign epithelium was nonreactive in 17/17 specimens without concurrent cancer (11 transurethral resection, 2 enucleation, 4 biopsy, p=0.002). Decreased NAG-1 expression in higher grade cancer is consistent with its known antitumorigenic, proapoptotic activities. PMID- 12894348 TI - Issues in the interpretation of breast core biopsies. AB - Core biopsies are now widely used for the nonoperative diagnosis of breast masses and microcalcifications and have replaced fine-needle aspiration (FNA) cytology for most lesions in many centers. In the United Kingdom, a scoring system of 5 reporting categories, B1-B5, has been adopted, which is to some extent similar to that used in the interpretation of breast FNA. This article is based on the practice at Charing Cross Hospital, London, which is a major regional breast screening and treatment center covering the West of London area; as well as on a thorough review of the contemporary literature. It begins by discussing issues related to the adoption of the technique and to handling core biopsies, followed by a brief presentation of the reporting categories. The article then deals with some commonly encountered diagnostic problems. PMID- 12894349 TI - GIST: an update. PMID- 12894350 TI - Myositis ossificans and fibroosseous pseudotumor of digits: a clinicopathological review of 64 cases with emphasis on diagnostic pitfalls. AB - Myositis ossificans (MO) and fibroosseous pseudotumor of digits (FP) are pseudotumoral mimics of malignancy. A review of 50 cases of MO and 14 cases of FP showed that a malignant diagnosis was suggested by referring pathologists in 23% of MO and 9% of FP. The most common misdiagnosis was osteosarcoma. Awareness of the spectrum of clinicopathological features of MO and FP will help pathologists avoid misdiagnoses. A comparison of the clinicopathological features of MO and FP showed that most features were similar, but FP involved an older age group (p<0.001). MO showed a statistically significant higher tendency to contain fibrinous material (p=0.007), edematous lymphangioma-like areas (p=0.013), and cartilage (p=0.017) and FP to contain excessive immature osteoid (p=0.029). These differences may be related to the site of occurrence. PMID- 12894351 TI - Correlation between histologic staging, hepatitis C virus genotypes and clinical features in HCV chronic hepatitis: evidence of a new pattern. AB - Genome heterogeneity may be related to the wide variability of clinical and pathological features in hepatitis C virus (HCV)-related chronic liver disease. This paper addresses the possible association between HCV subtypes and clinical and histological features of chronically infected patients. Sixty-eight consecutive liver biopsies of chronic hepatitis constituted the basis of the study. HCV genotyping was performed on frozen tissue. Grading of necroinflammatory activity and staging of fibrosis were histologically assessed. Serologic HCV-RNA and liver function were assessed at the same time. All information was compared with clinical data including age, sex, HCV serology, and probable data and route of infection. Two cases were excluded as inadequate tissue was available. Five cases were negative to HCV-RNA in both serum and tissue. In 61 cases HCV RNA was present at the same time in serum and liver tissue. Forty-four patients were men (72%) and 17 (28%) were women. Two peaks of age were observed: 1 in the 4th decade of life, the 2nd in the 7th. The 2 groups had different HCV genotypes. Patients with genotypes 1b (mean age 50.7 years), 2c (mean age 61.3 years), and a subgroup of coinfections (mean age 60 years) were older than patients with genotypes 1a (mean age 35.5 years), 3 (mean age 36 years), and a subgroup of coinfections (mean age 33 years). Patients with genotypes 1b, 2, or 2c and a subgroup of coinfections more frequently had a history of blood transfusion and or surgical intervention dating up to 49 years previously. Patients with HCV 1a, 3, and a subgroup of coinfections frequently admitted a period of intravenous drug abuse. Patients with advanced liver disease, i.e., severe fibrosis and cirrhosis, showed the same 2 peaks of incidence: in the 4th and 7th decades of life, the first group mainly comprising patients with HCV types 1a and 3, the second, patients with HCV types 1b and 2c. Both these groups shared a clinical history of a long-standing infection. Two profiles of patients emerged. The largest group was composed of elderly patients, infected by HCV genotypes 1b or 2c, with a history of blood transfusion and/or surgery, presenting an advanced stage of liver disease (namely, severe fibrosis or cirrhosis). The second group was composed of younger patients, mainly in the 4th decade of life, infected by HCV types 3 or 1a, often presenting with chronic hepatitis in the stage of severe fibrosis or cirrhosis. The latter could be the profile of HCV infection in the near future. PMID- 12894352 TI - Reactive neuroepithelial aggregates of the skin. AB - A peculiar, apparently reactive, microscopic structure of the skin was observed in 5 patients. It is characterized histologically by a bundle of nerve fibers partially or completely ensheathed by bland nonkeratinizing squamous epithelium. In 2 patients, the specimens were reexcision specimens, in which this structure was found amid active postbiopsy repair. In the remaining 3 patients, there was no history of previous excision or trauma at the lesional sites, and the background changes were consistent with either folliculitis or prurigo nodularis. This structure, when viewed in isolation, resembles carcinomatous perineural invasion. This structure and comparable structures observed in extracutaneous sites have often been referred to as "perineural invasion" in the literature, but the author proposes the term reactive neuroepithelial aggregate (RNEA) to denote its reactive nature. PMID- 12894353 TI - Assessing margins in breast tumors: a clay model. PMID- 12894354 TI - A mistake I made from which I learned much: melanoma in a preexisting congenital nevus of Unna's type. PMID- 12894355 TI - Images in pathology: mucinous tumor of the pancreas: corpora albicans-like structures within the ovarian stroma. PMID- 12894356 TI - Images in pathology: inclusion bodies in phyllodes tumor of breast. PMID- 12894357 TI - Images in pathology: baby faces: cross-sections of umbilical cords. PMID- 12894358 TI - Malignant monotypic epithelioid angiomyolipoma of the retroperitoneum. AB - Monotypic epithelioid angiomyolipoma (EAML) is an uncommon variant of angiomyolipoma, occurring primarily in the kidney and characterized by a predominance of HMB-45-positive epithelioid cells. EAML is generally considered a benign neoplasm, though rare cases exhibiting malignant behavior have been reported. We describe a patient with a primary retroperitoneal monotypic EAML, composed exclusively of atypical epithelioid cells, which metastasized to the liver and the mediastinum. This case represents, to our knowledge, the first description of metastatic disease occurring in an EAML of this particular site. PMID- 12894359 TI - Oncocytic angiomyolipoma of the kidney: a case report. AB - We describe a case of oncocytic angiomyolipoma composed of polygonal eosinophilic cells, that shares many features with renal oncocytoma. It was composed of alveolar structures, with accumulation of foamy histiocyte-like cells surrounding large globules of acellular, nonfibrillary, densely eosinophilic basal membrane like substance. Immunohistochemically, there was focal reactivity for HMB45 and negativity for the epithelial markers. Oncocytic angiomyolipoma lacks the atypical features of the recently described epithelioid variant of angiomyolipoma. None of the reported cases have been associated with tuberous sclerosis. PMID- 12894360 TI - Ductal adenocarcinoma of the pancreas with huge cystic degeneration: a lesion to be distinguished from pseudocyst and mucinous cystadenocarcinoma. AB - Cystic neoplasms of the pancreas are rare and often mistaken for pseudocyst by imaging studies and macroscopic examination. We describe an unusual tumor of the pancreas composed of a mural nodule of anaplastic carcinoma arising from a huge ductal adenocarcinoma undergoing cystic degeneration. The cyst measured 27 x 13 x 4 cm. Light microscopy showed that the cyst was partly lined by a single layer of cuboidal to columnar tumor cells with focal mucin production and was surrounded by hyalinized connective tissue. Most lining epithelial cells were absent owing to extensive degenerative process. Immunohistochemical studies showed positive staining of cytokeratin and vimentin for pleomorphic giant tumor cells, which were negative for leukocyte common antigen (CD45), KP-1 (CD68), epithelial membrane antigen (EMA), and carcinoembryonic antigen (CEA). The ductal adenocarcinoma stained strongly positive for cytokeratin and EMA, and negative for vimentin, CD45, CD68, and CEA. The clinical course of the current case was extremely poor and the prognosis resembled that of an anaplastic carcinoma. Therefore, we like to emphasize the importance of complete excision and extensive sampling of any cystic neoplasms in the pancreas including those with large cystic component to avoid missing the malignant elements. PMID- 12894361 TI - Upper gastrointestinal bacillary angiomatosis causing hematemesis: a case report. AB - A 38-year-old HIV-positive woman presented with massive hematemesis on initial admission to hospital. Endoscopy revealed ulcerated nodular lesions in the esophagus, stomach, and duodenum. The clinical impression was of Kaposi's sarcoma. The stomach was biopsied when the patient re-presented, and another endoscopy was performed. The biopsy showed mucosal ulceration with a proliferation of vascular channels associated with neutrophils and clumps of purplish, granular bacterial colonies, which were highlighted by a Warthin-Starry stain. The histopathological features were typical of bacillary angiomatosis. This case highlights bacillary angiomatosis involving the gastrointestinal tract at multiple sites, the cause of massive upper gastrointestinal hemorrhage that was the initial presentation of an HIV-positive patient, and the occurrence of visceral bacillary angiomatosis in the absence of cutaneous lesions. PMID- 12894362 TI - Nodular pulmonary amyloidosis following liver transplantation: report of a case and review of the literature. AB - Nodular pulmonary lesions seen in liver transplant recipients have a broad differential diagnosis including both infectious and noninfectious etiologies. Here, we report the first case of nodular pulmonary amyloidosis, an uncommon and benign localized form of amyloidosis occurring after orthotopic liver transplantation for end-stage primary biliary cirrhosis. PMID- 12894364 TI - Current status of intravascular stents as delivery devices to prevent restenosis. AB - The acute technical success of percutaneous transluminal angioplasty (PTA) has been improved with the use of intravascular stents. However, stent placement has led to the development of an increased myointimal hyperplastic response leading to late reduction in vessel lumen. Restenosis (> or =50% reduction in reference lumen diameter) rates for coronary angioplasty and stenting are reported between 20% and 50% at 1 year. Several studies are currently evaluating novel delivery of antiproliferative agents to prevent neointimal hyperplasia. The authors review the mechanism of neointimal hyperplasia as it relates to stent placement and discuss recent and ongoing trials evaluating intravascular brachytherapy and drug eluting stent technology in the inhibition of restenotic lesions. PMID- 12894365 TI - Clinical application of eversion carotid endarterectomy. AB - The results of 250 eversion carotid endarterectomies (ECEAs) in 227 consecutive patients in 1 institution were evaluated. The outcomes of 250 consecutive ECEAs at North Shore University Hospital by a single surgeon, between January 1998 and August 2001, were recorded prospectively. In the single series of 250 ECEAs the 30-day operative mortality was 0.4% and the perioperative stroke rate was 0.8%. During a mean follow-up of 23 months, the recurrent stenosis rate was 0.8%. A reduction in stroke and mortality rates is often observed with ECEA. PMID- 12894366 TI - Surgical intervention for acute intestinal ischemia: experience in a community teaching hospital. AB - The aim of this study was to evaluate the current management of acute mesenteric ischemia secondary to thrombotic or embolic occlusion of visceral vessels in a community teaching hospital. Between October 1997 and July 2000, a review of all hospital discharges revealed 83 patients with a discharge diagnosis of "acute vascular insufficiency-intestine." Among these 83 patients, 22 cases of acute mesenteric ischemia were confirmed. Management of these 22 patients was divided into 2 groups for analysis. In Group A, 14 patients were aggressively treated with visceral angiography (n=10), visceral artery bypass (n=8), visceral embolectomy (n=4), and bowel resection (n=7). In 8 of 14 of these patients, surgical intervention occurred in less than 24 hours from presentation. In Group B, 8 patients were managed with supportive care because of advanced age (mean age = 86 +/- 7 years), comorbid conditions, or patient and family preference. Postoperative morbidity in Group A consisted of cardiac events (n=3), pulmonary insufficiency (n=5), and prolonged gastrointestinal tract dysfunction (n=3). Twelve of 14 patients in Group A survived and were discharged, whereas only 2 of 8 patients in Group B survived and were discharged from the hospital. Although the literature suggests that there can be a significant delay in the diagnosis and treatment of acute mesenteric ischemia, the early recognition and aggressive treatment of acute mesenteric ischemia resulted in a good survival rate. Supportive management of very elderly and debilitated patients needs to be considered on a case-by-case basis. Although the outlook for such patients is dismal, survivors are possible as demonstrated by this series. PMID- 12894367 TI - Intraoperative thermographic monitoring during neurogenic thoracic outlet decompressive surgery. AB - This article reports the use of thermography to monitor 123 plexus decompressions for neurogenic thoracic outlet syndrome. The diagnosis and management of this disease continues to be controversial. Questions about pathologic mechanisms, the extent and frequency of muscular entrapment, scar, and interdigitations, as well as their relative contributions, remain. Thermographic visualization of the operated extremity allowed us to map and correlate thermal changes with specific surgical manipulations, as well as to analyze the tissues resected to better answer these questions. Initial thermal abnormalities indicating, usually, ulnar entrapments or irritation, normalized sequentially as discrete entrapments were resected. Thermographic monitoring continues to provide surgically useful information in one third of operations. PMID- 12894368 TI - Saratin, an inhibitor of collagen-platelet interaction, decreases venous anastomotic intimal hyperplasia in a canine dialysis access model. AB - Prosthetic dialysis access thrombosis and/or stenosis is the most common cause of graft impairment or loss and is primarily attributed to venous outflow stenosis due to intimal hyperplasia. Intimal hyperplasia is thought to result from interactions between areas of exposed subendothelial collagen in an injured vessel and platelets, resulting in platelet adhesion. Saratin, an inhibitor of the vWF-dependent binding of platelet to collagen interaction, has been shown in vitro to reduce the adhesion of platelets to collagen. In the current study, the authors investigated the effects of topical saratin administration in a canine dialysis access model in regard to intimal hyperplasia development at the venous anastomosis. Fourteen female mongrel dogs underwent placement of a femoral polytetrafluoroethylene (PTFE) dialysis access graft and were placed into 1 of 2 groups: 1) control or 2) experimental with topical saratin application. The experimental group had 600 microg of saratin (1 microg/microL) applied for 5 minutes directly onto the venous anastomosis before restoration of blood flow;control groups received vehicle control. At 4 weeks postoperative, a portion of the graft was removed along with a segment of the outflow vein. Veins were subsequently processed, sectioned, and analyzed along the length of the excised segment and divided into blocks that included the area of the graft toe, midanastomotic region and heel, and blocks A-E. Intimal hyperplasia was assessed by a computer-assisted morphometric analysis. Platelet counts and bleeding times were also measured. Vein segments in the control group (n=7) showed pronounced intimal hyperplasia in blocks B, C, and D as compared to the saratin group (n=6). Distribution of intimal hyperplasia by blocks between control and saratin groups were as follows: block [A] 8.6 +/- 1.9 vs 9.7 +/- 3.0% (p=NS), [B] 32.7 +/- 6.3 vs 10.7 +/- 3.5% (p=0.01), [C] 44.8 +/- 6.2% vs 10.3 +/- 1.5% (p=0.0004), [D] 40.8 +/- 11.0 vs 9.1 +/- 4.2% (p=0.02), [E] 7.5 +/- 5.5 vs 2.7 +/- 0.4% (p=NS). Intimal hyperplasia normalized to vein wall thickness also showed a significant reduction with saratin application. Bleeding times and platelet counts obtained at different time points during the experiment showed no difference between control and saratin groups. In a canine dialysis access model using PTFE grafts, topical application of saratin at the venous anastomosis decreased intimal hyperplasia development by as much as 77% when compared with control animals. Saratin provides for a method of substantially reducing intimal hyperplasia by direct local application without systemic side effects. PMID- 12894369 TI - Primary utilization of stents in angioplasty of superficial femoral artery. AB - Percutaneous transluminal angioplasty (PTA) of infrainguinal arteries has been frequently reported in the literature. Independent of the technical feasibility, the results, in short-and long-term follow-up differ among published series. The objective of this article was to assess the long-term results of angioplasty in small, (<3 cm) segmental lesions of the superficial femoral artery treated with primary stenting. Eighteen patients (12 men, 6 women) with lesions of the superficial femoral artery smaller than 3 cm were selected for endovascular treatment and follow-up. The median patient age was 65 years (range, 54 to 84). The indication for treatment was intermittent claudication in 7 patients and critical ischemia (ischemic pain associated with trophic lesions) in 11 patients (62%). Of these, 13 lesions were stenoses and 5 total occlusions. The run-off was good in 15 patients who had more than 2 distal arteries and 3 had only 1 patent artery. During follow-up, all patients were observed with physical examination, progressive exercise treadmill test; pulse volume was measured with Doppler and duplex scan after 1, 3, and 6 months, and then twice a year. There was only 1 primary failure; the initial success rate was 94%. During follow-up of 40 months (6 to 70 months), only 1 patient presented with thrombosis of the stent 6 months after the procedure. There was no symptomatic restenosis in our study. One patient suffered an acute myocardial infraction after 45 months and died. The primary patency after 1 and 3 years was 88%. No patient was lost to follow-up. Primary stenting of segmental lesions (less than 3 cm) of the superficial femoral artery produced satisfactory results immediately and during long-term follow-up. PMID- 12894370 TI - Endovascular treatment of innominate artery stenosis in a bovine aortic arch--a case report. AB - Recent reports have established the feasibility and safety of percutaneous transluminal angioplasty and stent placement in the treatment of innominate artery occlusive disease. Although the long-term durability of these endovascular approaches has not been clearly established, they are particularly attractive in patients who are not considered good candidates for surgical reconstruction. The authors present a case involving a morbidly obese Hispanic woman who had undergone previous coronary artery bypass and complained of right visual symptoms, dizziness, and right upper extremity claudication. Because of her unusual bovine anatomy and the desire to avoid reoperating in her chest, a unique approach with a left subclavian-to-carotid bypass and innominate artery angioplasty and stenting was used. PMID- 12894371 TI - Small ruptured abdominal aortic aneurysm with renal failure: endovascular treatment--a case report. AB - The successful endovascular exclusion of a ruptured 3-cm diameter atherosclerotic abdominal aortic aneurysm (AAA) in a high-risk patient with renal failure is reported. An 82-year-old man with chronic renal failure and other comorbidities was admitted for acute abdominal pain. Duplex scan and computed tomography showed a ruptured 3-cm diameter atherosclerotic AAA. As a consequence of the patient's high surgical risk combined with signs of rupture, despite the progressively decreasing renal function, an emergency exclusion of the AAA was performed by means of a bifurcated Excluder (W. L. Gore and Associates) endovascular graft. The procedure was performed by minimizing administration of iodinated contrast medium using a guidewire into the lowest renal artery as a marker of proximal deployment. Intravascular ultrasound was used to confirm correct deployment. The postoperative recovery was characterized by acute renal insufficiency and bowel ischemia, which were treated with ultrafiltration and medical therapy, respectively, with complete resolution. PMID- 12894372 TI - Isolated abdominal aortic rupture in a child due to all-terrain vehicle accident- a case report. AB - Rupture of the abdominal aorta as the result of blunt trauma is uncommon, due in part to its protected position in the retroperitoneum. Isolated aortic rupture following blunt injury is even more rare. A case of isolated abdominal aortic rupture in a 6-year-old girl following blunt trauma due to an all-terrain vehicle accident is reported. The girl survived following an emergent aortoiliac reconstruction. PMID- 12894373 TI - Mesenteric venous thrombosis in a patient with prothrombin 20210A mutation and antithrombin III deficiency: challenges to conventional anticoagulation--a case report. AB - Mesenteric venous thrombosis presents as vague abdominal pain in patients with a medical or family history suggestive of a hypercoagulable state. Classic computed tomography findings will often confirm the diagnosis, and the presence of persistent abdominal pain or tenderness will determine the need for surgical intervention. Expeditious anticoagulation is the treatment of choice. This case demonstrates the CT findings for mesenteric venous thrombosis and discusses the challenges of anticoagulation in a patient with 2 hypercoagulable disorders. PMID- 12894374 TI - Laparoscopic exclusion of a splenic artery aneurysm--a case report. AB - Splenic artery aneurysm is a rare but serious vascular disease. The mortality risk is 36% when one is ruptured. Surgical therapy has traditionally consisted in resection through a laparotomy. The authors' experience of a case of laparoscopic exclusion of a splenic artery aneurysm is reported. This surgical approach is simple, safe, and minimally invasive. This procedure should increase the indication for surgical treatment of a splenic artery aneurysm. PMID- 12894378 TI - Basic principles of immunotherapy for neurologic diseases. AB - Immunosuppressive drugs and immunomodulating procedures can improve the quality of life in patients with immune-related neurologic diseases and can even be lifesaving if properly used to avoid deleterious or irreversible adverse effects. For the successful use of these drugs or procedures, the treating physician must be familiar and comfortable with the drugs' mode of action, the accuracy of the diagnosis, the goals and expectations of therapy, and the risk-benefit ratio of the therapy. Improvement must be based on a beneficial change in activities of daily living and not solely on a change in the laboratory test values. PMID- 12894379 TI - Current disease-modifying therapies in multiple sclerosis. AB - In recent years, the usefulness of interferon beta and glatiramer acetate in the treatment of relapsing-remitting multiple sclerosis (RRMS) has been established. Interferon beta has also been shown to be efficacious in secondary-progressive multiple sclerosis (SPMS) as well as in patients with isolated syndromes at risk to develop clinically definite multiple sclerosis (MS). Mitoxantrone is another disease-modifying drug that is available for SPMS and severe cases of RRMS. The clinical utility of disease-modifying agents in MS will be reviewed with respect to the anti-inflammatory, immunomodulatory, and immunosuppressive treatments that are currently available. Symptomatic therapies will not be considered. PMID- 12894380 TI - Future immunotherapies in multiple sclerosis. AB - Immunotherapy of multiple sclerosis (MS) will continue to benefit from an increasing understanding of this disease. This knowledge results in newly defined targets for novel therapies. In this article the development of future immunotherapies will be discussed by classifying the approaches into three main types: (1) antigen-specific therapies; (2) agents with a defined target in pathogenic steps of the MS lesion; and (3) therapies with broad immunomodulatory activity. Success in developing new immunotherapies depends on understanding the underlying complexity and heterogeneity of the disease. The current practice of employing a single therapy across a heterogeneous group of MS patients is in part a likely reason for their modest efficacy. The mechanism of action of a single agent may target the appropriate defect in one individual but not others. The therapy of MS in the future will most likely use a combination of agents that are directed at the underlying disease state and stage in the individual patient. PMID- 12894381 TI - Therapeutic strategies in the Guillain-Barre syndrome. AB - The Guillain-Barre syndrome (GBS) represents the prototypic immune-mediated peripheral neuropathy, which is now recognized as a group of conditions with diverse pathology and pathogenesis. The observation that plasma exchange and intravenous immunoglobulin result in clinical improvement, the presence of circulating antibodies targeting structures on peripheral nerve tissue in sera from GBS patients, and the deposition of immunoglobulins and complement demonstrated on myelinated fibers in nerve biopsies from affected patients point to a critical role of the humoral immune response in the pathogenesis of GBS. However, the observation of inflammatory infiltrates in nerve and the critical pathogenic role of neuritogenic T lymphocytes as demonstrated in the animal model support the concept that a disordered cellular immunity is also of critical importance in the pathogenesis of this neuropathy. Current treatment strategies are aimed at mitigating the harmful effects of the immune system on peripheral nerve. This review will address the rationale for immunotherapy in GBS based on experimental and immunologic studies of the pathogenesis of this disease. PMID- 12894382 TI - The treatment of chronic inflammatory demyelinating polyradiculoneuropathy. AB - Chronic inflammatory demyelinating polyradiculoneuropathy is an inflammatory disorder of nerve that usually presents with slowly progressive weakness and sensory loss and areflexia. It is among the most treatable of the peripheral nerve disorders, and several modalities have been shown to be effective in prospective, randomized controlled trials. Although most patients show a gratifying early response to treatment, in many cases the patients relapse. The cumulative effects of the neuropathic impairments, along with side effects from long-term immunosuppressive treatment, combine to produce significant long-term morbidity and loss of function. This review will cover the epidemiology, clinical features, laboratory findings, and pathogenesis of chronic inflammatory demyelinating polyradiculoneuropathy; the current status of the treatment of this disorder will be reviewed, highlighting those therapies shown to be effective in randomized controlled trials. PMID- 12894383 TI - Antibody-associated polyneuropathy syndromes: principles and treatment. AB - Treatment of immune-mediated neuropathies first requires an accurate diagnosis. The diagnosis is often based on clinical, electrophysiological, and immunological features of the syndrome. The selection of appropriate therapies is then based on the spectrum of response of a syndrome to medications and an assessment of possible side effects. In neuropathies with associated serum immunoglobulin M autoantibodies, such as anti-myelin-associated glycoprotein and motor syndromes, the choices of therapy are often limited to cytotoxic agents and, in some cases, intravenous immunoglobulin. In neuropathies with immunoglobulin G antibodies in both serum and cerebrospinal fluid, such as sensory neuronopathies associated with anti-Hu antibodies, there is no well-documented response to any immunotherapy. The general principles regarding therapy of immune neuropathies will be discussed with a focus on diagnosis and treatment options of the demyelinating and immunoglobulin M antibody-associated neuropathies. PMID- 12894384 TI - Therapy in myasthenia gravis and Lambert-Eaton myasthenic syndrome. AB - Myasthenia gravis (MG) is a heterogeneous disorder, a fact that needs to be kept in mind when considering treatment. Most patients benefit from pyridostigmine. In nonthymomatous ocular MG, prednisolone is often effective. Thymectomy is indicated for thymoma and is an option for acetylcholine receptor antibody positive patients with generalized weakness developing under the age of 45 years. In older patients and in those failing to respond to thymectomy, prednisone alone or combined with azathioprine is the treatment of choice. Mycophenolate mofetil is an option in those intolerant of azathioprine. Lambert-Eaton myasthenic syndrome (LEMS) can exist in paraneoplastic (P-) and nonparaneoplastic (NP-) forms. Most patients benefit from 3,4-diaminopyridine. In P-LEMS, treatment of the tumor often results in neurological improvement. In both forms, prednisone alone is an option or combined with azathioprine in NP-LEMS. In both MG and LEMS, where weakness is severe, plasma exchange or intravenous immunoglobulin treatment may provide short-term benefit. PMID- 12894385 TI - Therapeutic approaches in patients with inflammatory myopathies. AB - Among the group of inflammatory myopathies, dermatomyositis (DM) remains the most treatable subset responding, in the majority of the cases, to steroids, intravenous immunoglobulin (IVIg), or immunosuppressants. Inclusion-body myositis (IBM) remains the most difficult disease to treat; in uncontrolled studies immunosuppressants and steroids have not helped, and controlled trials with IVIg have been disappointing. Polymyositis (PM) is a very uncommon, although still overdiagnosed, disorder and its rarity poses difficulties in performing large scale therapeutic studies; based on small series, however, PM seems to variably respond to immunotherapeutic interventions. The most consistent problem in the treatment of inflammatory myopathies remains the distinction of true PM from the difficult-to-treat cases of IBM, or from necrotizing myopathies and dystrophic processes where secondary endomysial inflammation may be prominent. The future in the management of PM, DM, and IBM seems promising because of the availability of new agents directed at T-cell activation molecules, cytokines, chemokines, and adhesion receptors. In IBM, the use of such immunomodulatory drugs may be combined with agents that block cytokine-enhancing amyloid or with agents that inhibit the formation and polymerization of amyloid fibrils. PMID- 12894386 TI - Therapy of neurological disorders in systemic vasculitis. AB - Significant progress has been made in our understanding of the underlying cellular and molecular mechanisms associated with vasculitis. Simultaneous involvement of the nervous system and systemic organs points to shared antigenic targets or immune mechanisms operative at both sites. Although specific immunotherapy is not available, the old and new immunomodulatory therapies have been instrumental in providing effective therapy in most of these chronic autoimmune disorders. The treatment of giant cell arteritis, neurosarcoidosis, Wegener's granulomatosis, systemic lupus erythematosus, polyarteritis, Sjogren's syndrome, and Behcet disease will be reviewed. PMID- 12894387 TI - Paraneoplastic neurologic syndromes: approaches to diagnosis and treatment. AB - There is increasing evidence that many paraneoplastic neurologic syndromes are immune-mediated. Because symptoms usually have an acute onset and precede the tumor diagnosis, the recognition of a neurologic disorder as a paraneoplastic syndrome is challenging. The poor response to treatment of many paraneoplastic neurologic syndromes has been attributed to the early, irreversible pathological changes involving the nervous system. This may account for the more frequent responses to therapy of some paraneoplastic neurologic syndromes affecting the peripheral nerves and neuromuscular junction compared with those that produce neuronal degeneration. This review focuses on the current therapeutic approaches to immune-mediated paraneoplastic neurologic syndromes, including prompt recognition of the disorder, detection and treatment of the associated tumor, and, in some disorders, immunosuppressive therapy. PMID- 12894389 TI - [The DMW scores thrice]. PMID- 12894390 TI - [The quality of medical education in the print media]. PMID- 12894391 TI - [New nondispersive infrared spectrometry in 13C-urea breath tests]. AB - BACKGROUND AND OBJECTIVE: 13C-urea breath tests have become clinical routine in the diagnosis of Helicobacter pylori infection. For the analysis of the 13CO2/12CO2 enrichment in breath, less expensive alternatives to the expensive mass spectrometry (IRMS) have been developed, based on isotope-selective infrared spectroscopy (NDIRS). In this prospective study we tested under clinical conditions a simplified and thus less expensive NDIR-spectrometer by comparing it with mass spectroscopy. METHODS: 100 patients (53 men, 47 women, mean age 59+/-14 years) with dyspeptic symptoms were tested for Helicobacter pylori infection using the 13C-urea breath test. The isotope ratio analysis of the breath samples was performed in duplicate, both using IRMS and NDIRS. RESULTS: The results of the baseline-corrected 13CO2 -exhalation values between IRMS and NDIRS were in excellent agreement. The mean difference between both methods was 0.05+/-1.16 . Evaluating the qualitative urea breath test results in reference to IRMS as the reference, the NDIRS had a sensitivity of 95 % and a specificity of 99 %. CONCLUSION: This newly developed isotope-selective nondispersive infrared spectroscopy is going to become a reliable, and low-cost alternative to expensive isotope ratio mass spectrometry in the analysis of 13C-breath tests. All these characteristics make NDIRS particularly suitable for laboratories where the daily number of assays is small or for use in the doctor's office PMID- 12894392 TI - [Rhabdomyolysis, hyponatremia and fever in a patient with Schmidt's syndrome]. AB - HISTORY: A 38-year-old man had fever (40 degrees C) and a swollen right leg. Two weeks before admission he had received non-steroid anti-inflammatory drugs (paracetamol and ibuprofen) after a tooth extraction. Some weeks before he had noticed a rough voice and a dry skin. INVESTIGATIONS: The patient had a sinus tachycardia of 145/min (blood pressure of 130/80 mm Hg). Laboratory data revealed a CK of 16,650 U/l (< 80 U/l), myoglobin of 2420 U/l (< 90 microg/l) and LDH of 1250 U/l (<240 U/l), leukocyte count of 12,000 / microl and C-reactive protein of 3.0 mg/dl (<0.5 mg/dl). Sodium was markedly decreased (110 mmol/l (135 - 145 mmol/l)). Determination of thyroid hormone showed primary hypothyroidism with an elevated TSH of 62.6 mU/l (0.3 - 4.0 mU/l); T4 of 38 nmol/l (58 - 154 nmol/l), T3 1.17 of nmol/l (1.23 - 3.08 nmol/l) and fT4 of 6 pmol/l (10 - 25 pmol/l). The thyroid autoantibodies were increased (thyroxine peroxidase antibodies of 684 U/l (<35 U/l) and thyroglobulin antibodies of 173 U/l (<40 U/l)). Ultrasound of the thyroid revealed an nonhomogeneous structure. Cortisol at 8.00 a. m. was reduced by 63 mmol/l (180 - 640 mmol/l) and did not increase after administration of ACTH (60 min. cortisol at 90 mmol/l (>550 mmol/l)). ACTH was increased (141 pg/ml; normal range 17 - 52 pg/ml). TREATMENT AND COURSE: The initial therapy consisted of hydrocortisone (100 mg i.v as bolus and 100 mg during the next 24 hours) and levothyroxine replacement (200 micro g) was initiated. During the following 8 days clinical symptoms regressed. Values of sodium, myoglobin and LDH decreased. After therapy with cephalosporin (Ceftriaxon) and penicillin (Flucloxacillin) fever and inflammation parameters decreased. CONCLUSION: This is a rare case of a rhabdomyolysis and hyponatriaemia due to hypothyroidism and Addison's disease (Schmidt's syndrome). PMID- 12894393 TI - [Cutaneous lesions in lupus erythematosus]. PMID- 12894394 TI - [Cardiovascular risk factors in women]. PMID- 12894395 TI - [Indications for the use of immunoglobulin therapy]. PMID- 12894396 TI - [Praeimplantation genetic diagnosis: arguments from a paediatric point of view]. PMID- 12894397 TI - [Which treatment for cartilage regeneration is recommended in arthrosis?]. PMID- 12894398 TI - The human parietal cortex: a novel approach to its architectonic mapping. PMID- 12894399 TI - Magnetic resonance imaging study of the parietal lobe: anatomic and radiologic correlations. PMID- 12894400 TI - Subcortical projections of the parietal lobes. AB - Parietal lobe visual areas project to several subcortical targets. Three of the most prominent of these are to the basal ganglia, superior colliculus, and pontine nuclei. All three are probably involved in some aspect of visually guided movement. One of the largest systems of fiber connections in the human brain arises in the cerebral cortex, and relays to the cerebellum via the pontine nuclei. Pontocerebellar axons terminate within the cerebellar cortex as mossy fibers. Cells in the cerebral cortex that project to the pontine nuclei are all pyramidal in shape, and all are located in lamina V of the cerebral cortex. Subcortical projections from the somatosensory cortex of rats shows that cells that project to the basal ganglia and cerebellum occupy different sublaminae of layer V. Some cortical pyramidal cells' axons may divide, with one branch projecting to the superior colliculus, the other to the pontine nuclei. Temporal lobe visual areas, which are involved in form recognition and learning, do not project to the cerebellum. Reciprocal connections from the cerebellum back to the parietal lobe may play a role in converting visual information from retinotopic to head-centered coordinates. PMID- 12894401 TI - The organization of somatosensory cortex in anthropoid primates. PMID- 12894402 TI - Is there a role for the parietal lobes in the perception of pain? AB - Converging lines of evidence confirm a role for the anterior parietal cortex in pain processing and extend the traditional view of SI to include discriminative aspects of somatic stimulation that is potentially tissue-damaging (e.g., painful). Recent studies more specifically implicate SI in the sensory aspect of pain perception by demonstrating that SI activation is modulated by cognitive manipulations that alter perceived pain intensity, but not by manipulations that alter unpleasantness, independent of pain intensity. Nevertheless, despite the probable role of SI in the encoding of the various sensory features of pain, considerable evidence suggests that nociceptive input to SI may also serve to modulate tactile perception. Thus, SI cortex may be involved in both the perception and modulation of both painful and nonpainful somatosensory sensations. Defining a role in pain processing for the parietal operculum is somewhat more problematic. The absence of a fine somatotopic organization of cutaneous (or visceral) receptors virtually eliminates a substantial role for this region in localizing noxious stimuli. Several studies suggest separate representations for pain and touch within the posterior parietal cortex and SII, respectively; however, inter-species differences in cortical anatomy and inconsistencies in the designation of SII proper preclude a clear reconciliation of the data. Likewise, suggestions that SII activation is predominantly related to processing the nociceptive quality of the stimulus (60,61) are inconsistent with many studies in both human and nonhuman subjects, which show a strong functional relationship between SII activity and innocuous (especially, vibrotactile) stimulation. Nevertheless, the numerous studies indicating pain related activation within the parietal operculum (and/or SII) underscore the potential importance of this region in the perception of pain and the need for continued research. Finally, a possible role of posterior parietal cortex (BA 5/7, 39/40) in orientation and attention toward painful sensory stimuli is consistent with existing literature describing this region as a poly modal association area concerned with intrapersonal and extrapersonal space; however, results from studies that actually manipulate the subjects' level of attention relative to painful stimuli have not uniformly supported this hypothesis (75). Future studies assessing both attentional demand and direct manipulation or motor interactions involving noxious stimuli may help to resolve this issue. In spite of some discrepant results concerning specific details of the nociceptive process, the weight of human pain research now firmly establishes a role for the parietal lobes in the conscious appreciation of the sensation of pain. PMID- 12894403 TI - Anatomic and functional reorganization of somatosensory cortex in mature primates after peripheral nerve and spinal cord injury. PMID- 12894404 TI - Mechanisms of selection and guidance of reaching movements in the parietal lobe. PMID- 12894405 TI - The role of the parietal cortex in grasping. PMID- 12894406 TI - The role of the parietal cortex in the neural processing of saccadic eye movements. AB - The lateral intraparietal area has a signal that describes a saccade target, maintains the memory of a saccade plan during a delay, and describes the saccade itself. It is unlikely, however, that this signal generates a plan for the saccade, because most neurons with this delayed saccade activity also respond, sometimes more strongly, to salient stimuli that are unlikely to be saccade targets. Instead, it is more likely that this saccadic signal performs two functions unrelated to saccade planning itself. The first function is to contribute to a salience map: it is well known that attention is located at the goal of a saccadic eye movement, and recent experiments detailed here show that the attentional advantage of the saccade goal is [figure: see text] maintained throughout the delay period of a memory-guided saccade. The saccade signal, presumably driven by the frontal eye fields or other prefrontal cortical areas, informs the salience map of a saccade plan, and therefore renders the goal of the saccade a salient location for attentional processes and, possibly, to provide targets for future saccades. The second function is to use the saccade signal to provide information by which the parietal cortex can update the visual representation to compensate for an eye movement, thus maintaining a spatially accurate vector map of the visual world despite a moving eye. PMID- 12894407 TI - Sensorimotor integration in posterior parietal cortex. PMID- 12894408 TI - Somatosensory and motor disturbances in patients with parietal lobe lesions. AB - Lesion studies show that a wide range of integrative sensorimotor functions can be selectively disturbed in patients with parietal lobe damage. Lesions restricted to the somatosensory representations on the anterior parietal lobe produce somatosensory deficits that resemble deafferentated states, including the secondary effects on motor control. Slightly more posterior lesions often are associated with impairment of more complex synthetic somatosensory functions similar to those observed after dorsal column lesions. Damage of the posterior parietal lobe can selectively interfere with virtually every aspect of somatosensory function. These perceptive and cognitive disturbances may or may not be associated with complex motor disturbances of the apractic type. The frequent association of astereognosia and tactile apraxia illustrate the mutual interdependence of the sensorimotor processes involved in active touch and the proximity of the respective processing modules. Parietal lobe function is critical for the control of force and posture, and for the formation of the body image and its relation to external space (the guidance of movements, including the eyes, to external objects). Imaging studies underscore the prominent role of the parietal cortex as a sensorimotor interface and provide complementary information about the interrelationship between perception and action. Action observation activates premotor cortex, but parietal cortex is also recruited whenever an action involves objects, thus emphasizing the significance of parietal cortex for object-directed motor behavior. In contrast to the ventral dorsal route processing dichotomy in the visual system, both the perceptual cognitive and motor aspects of somatosensory processing are compromised after parietal damage, demonstrating a different functional architecture of the two sensory systems. The preservation of the comprehension of the meaning of gestures or of object use in patients with lesions restricted to the parietal lobe reveals that the semantic aspects of motor behavior are mediated in the temporal lobe. PMID- 12894409 TI - Remapping somatosensory cortex after injury. AB - The results reported here show convincingly that the adult somatosensory cortex alters its maps subsequent to injury. Studies with amputees and chronic pain patients have shown that pain may be one important perceptual correlate of the changes that were observed in primary somatosensory cortex. These results also have led to new approaches to rehabilitation. Both pharmacologic and behavioral interventions designed to alter cortical reorganization were found to not only alter the organization of primary somatosensory cortex but also maladaptive perceptual phenomena that accompany these changes. PMID- 12894410 TI - The visuomotor functions of posterior parietal areas. PMID- 12894411 TI - Multimodal spatial representations in the human parietal cortex: evidence from functional imaging. PMID- 12894412 TI - Apraxias and the lateralization of motor functions in the human parietal lobe. PMID- 12894413 TI - Interactions between the dorsal and ventral streams of visual processing. PMID- 12894414 TI - Parieto-frontal networks and gaze shifts in humans: review of functional magnetic resonance imaging data. PMID- 12894415 TI - Modular organization of parietal lobe functions as revealed by functional activation studies. PMID- 12894416 TI - Neglect syndromes: the role of the parietal cortex. PMID- 12894417 TI - Hemispheric asymmetries in the parietal lobes. PMID- 12894418 TI - Parietal lobe epilepsy. PMID- 12894419 TI - Surgical management of parietal lobe epilepsy. AB - Parietal lobe seizure foci are difficult to localize unless there is an MRI lesion or contralateral sensory aura. Rapid network projection often makes scalp EEG and semiology misleading. However, seizure control can be achieved with reasonable success when concordant information guides the physician to a parietal ictal onset. Perhaps the most important messages that this small surgical series provides is that of neurologic outcome. The parietal lobe is a highly convergent cortical region and a major network way station. Except for primary sensory phenomena and language, one cannot temporarily ablate parietal cortical association area within a presumed epileptogenic region and predict the visuospatial, cognitive, and neurologic outcome. Therefore, data demonstrating that one can resect regions of parietal cortex and not cause serious dysfunction are helpful. The mild morbidity encountered in this group of patients would not be necessarily predicted if the same region of normal parietal lobe was resected. Therefore, one must consider cortical plasticity and functional redistribution as possible reasons for this, particularly when most of these substrates are of developmental origin. PMID- 12894420 TI - [Factors influencing the hormonal function in patients after the treatment of larynx cancer]. AB - It has been revealed that the hypothyreosis (subclinical as a rule) can be found after every method applied in treatment of the larynx cancer. Correlation of data obtained with the time function allowed to specify survival probability of patients in euthyreosis after therapy termination. Among the factors supposed to be involved in keeping patient's normal thyroid gland function the following have been taken into account: sex, age, stage of the neoclassic process, scope of the surgical intervention on the larynx and lymph nodes of the neck, thyroid gland surgery and dose of irradiation given in Gy/T at the time of radiotherapy. Substantial impact on hypothyreosis was attributed to: method of laryngeal cancer, treatment, the range of its structures removal and surgical intervention on thyroid gland (excision of the thyroid lobe or its translocation for larynx reconstruction). It was found that the probability of survival in euthyreosis in patients treated by surgery and by combined method was the least after laryngopharyngectomy. In 3 years follow up period after such a therapy it amounted to 75% for surgical method and 48% for combined one. It was evident that the dysfunction of the thyroid gland resulted mostly from combined method of laryngeal cancer therapy (even without surgical intervention on thyroid glad). The probability of survival in euthyreosis in those patients was the least of all and amounted to 78.7% after 1 year of therapy termination and 42.9% after 6. In patients treated with combined method and intervention on thyroid gland, the survival probability in euthyreosis was 25% after excision thyroid lobe and 20% after its translocation for larynx reconstruction. After exclusive surgical therapy the probability amounted to 96% and after radiotherapy 88.6%. Prolongation of the follow up after radiotherapy (over 1 year) didn't increase the number of patients with hypothyreosis. The necessity of the evaluation of the thyroid gland hormonal function in every one patient has been stressed. This should be done before treatment and 1, 6, and 12 months after the termination of the therapy, and then once a year. PMID- 12894421 TI - [Patients under 45 years of age "young adults" with head and neck squamous cell carcinoma. Retrospective, multivariable analysis--preliminary report ]. AB - There is a clear tendency to diagnose squamous cell carcinoma of the head and neck more frequently in patients below 45 years of age. These neoplasms in "young adults" seem to be more aggressive with rapid progression, frequent reappearance after treatment and worse prognosis than is observed in older patient population. This preliminary report presents results of retrospective, multivariable analysis of patients below 45 years of age with squamous cell carcinoma of the head and neck. The special focus of the study were: disease progression (T), regional node and distant metastases (N,M), tobacco and alcohol consumption, environmental factors, concomitant diseases, kind of surgical technique and tumour histology. PMID- 12894422 TI - [Gastro-oesophageal reflux disease presenting as laryngeal pathology- epidemiology, symptoms, diagnosis. Preliminary report]. AB - Symptoms from the ears, throat, nose and larynx--the laryngeal mask's manifestations of gastroesophageal reflux disease (GERD) are important clinical problem of significant occurrence. AIM: (1) Determination of GERD's frequency of occurrence, especially within the group of patients with the pathology of the posterior laryngeal commissure. (2) Evaluation of the effective algorithms for detecting and management of patients demonstrating symptoms of laryngeal mask, which develop on the ground of gastroesophageal reflux disease. The investigation included 9 patients who were hospitalized in the Department of Otolaryngology of Karol Marcinkowski University of Medical Sciences in Poznan. Every patient was carefully interviewed by means of 3 questionnaires: the first one being Karlsson's general questionnaire, the second one--author's questionnaire designed in the Department of Otolaryngology of Medical Academy in Poznan which emphasises laryngeal symptoms; and the third one being the inquiry including serious past diseases, regularly taken medications, and stimulants. The objective condition of the hypopharynx and larynx was estimated on the basis of indirect laryngoscopy, followed by manometric examination of the oesophagus and 24-hour pH-metry carried out in Motility Laboratory of Surgical Department. 8 patients (88.9%) out of 9 were diagnosed as having GERD on the ground of the questionnaires as well as on the base of the results from the 24-hour pH-metry, and among 7 patients (87.5%) out of the 8 motility disfunction of the oesophagus was detected. Within the group of 8 patients with diagnosed GERD on the ground of additional tests 3 patients (37.5%) were diagnosed as having posterior laryngitis, 2 patients (25%) were diagnosed as having chronic laryngitis and the last 1 (12.5%) suffering from dysphagia. Among 3 patients (37.5%) with tumors of the larynx of rare location within the posterior laryngeal commissure coexistence of gastroesophageal reflux disease (GERD) was proved. The remaining 1 patient (11.1%) with the larynx cancer had no confirmation of GERD. The connection between chronic laryngitis and gastroesophageal reflux disease and/or motility disfunction of the oesophagus described in the literature was confirmed within the group of examined patients. Awareness of GERD and diagnostics carried out in that direction is needed in the group of patients with typical symptoms of the laryngeal mask. Significant frequency of coexistence of gastroesophageal reflux disease and pathology of the posterior laryngeal commissure requires effective algorithms for management of these patients. PMID- 12894423 TI - [Angiofibroma juvenile in the clinical material of Department of Otolaryngology in Poznan in the years 1977-2000 ]. AB - 30 patients suffering from angiofibroma juvenile were treated in the Department of Otolaryngology in Poznan in the years 1977-2000. The patients aged from 9-36 years. All the patients were treated surgically and the method of choice was Denker's surgery. 3 patients were reoperated because of local recurrence. On the base of own material good results of full recovery of patients with angiofibroma treated surgically are presented. PMID- 12894424 TI - [Difficult therapeutic decisions in clinically advanced head and neck tumors--the role of quality of life factors]. AB - Among the patients with IV stage advancement of head and neck tumours, the group selected to radical treatment and group primary treated with palliative intention are nearly equal. The factors which are, or should be taken into consideration in difficult therapeutic decisions concerning this patients, has been critically discussed. The quality adjusted survival score, factor of opportunity cost, the real place of patient in critically difficult therapeutic decisions, the doctors attitude to this problem, and the potential risk of abusing the quality of live factor in decision considering the euthanasia. PMID- 12894425 TI - [Remote results of laryngeal cancer treatment T1-2N0M0]. AB - We have analyzed results of special treatment 187 patients with cancer of a larynx T1-2N0M0. The patients were parted into four clinical groups. 45 patients of the first group have received combined treatment, including a partial resection of the larynx and preoperative radial therapy in a general dose up to 40 Gr. 47 patients of the second group have received a course of the radial therapy under the radical program in total dose 70 Gr. 63 patients of the third group have passed a course of combined treatment, including the partial resection of the larynx and pre- and postoperative radial therapy in total dose, equivalent 60 Gr of a standard procedure. 32 patients of the fourth group have passed the extremely surgical treatment (partial resection of the larynx). The obtained results speak about large efficiency of combined treatment. As contrasted to with radial therapy under the radical program quantity of relapses considerably has decreased in the field of the primary center. As contrasted to with surgical treatment statistically quantity of recurrence of disease in area of regional lymphatic nodes authentically has decreased. PMID- 12894426 TI - [Significance of support groups for patients after total laryngectomy for laryngeal carcinoma]. AB - The aim of this study was to determine the relationship between the quality of life (QL) assessment and attachment to support group in patients after total laryngectomy. Material consisted of 60 patients with squamous cell carcinoma of the larynx, inclusive of 30 patients belonging to the supported group. The questionnaire of QL EORTC QLQ-C30, the EORTC head and neck cancer module QLQ H&N35 and Hospital Anxiety and Depression Scale HAD were used. We found out that the somatic, performance and social dimension of QL of patients belonging to the supported group are higher than others patients. PMID- 12894427 TI - [Connection between changing the vitamin and immune status and the character of the throat microflora in patients with chronic tonsillitis]. AB - The author investigated 28 patients with chronic tonsillitis. The fungi sort Candida on the tonsils was discovered at 10 (35.72) patients, the bacterial flora was discovered at the other patients. There were more considerable decrease content of vitamins B2, C, methylnicotinamide and methylmalonic acid in the urine, vitamins B2 and methylnicotinamide in the saliva, vitamins A and B1 in the serum of the blood in the group of the patients with the fungus flora on the tonsils. It was discovered the decrease of numbers of lymphocytes CD8 and level of immunoglobulins G and M, disimmunoglobulinemia in this group. This decrease of the vitamins, cell and humoral immunity level leads to disbacteriosis, decompensation of the disease and development of the complications. PMID- 12894428 TI - [Resection of tracheal stenosis with end to end anastomosis]. AB - The aim of the study was evaluation of early results of tracheal resection with end to end anastomosis. Tracheal resection was performed in 5 patients with tracheal stenosis resulting from prolonged intubation. Three patients were admitted without tracheostomy, in two patients tracheostomy was performed in another hospital. Diagnostic tracheoscopy was performed in all the patients and the tracheal stenosis was found in a distance of 2.5-4 cm from the vocal cords. The diameter of stenosis was 1-4 mm, and the length of stenosis was as long as 3 5 cartilages of trachea. Three cartilages were resected in one patient, 4 cartilages in two patients and 5 cartilages in two patients. End to end anastomosis was performed using absorbable sutures (Vicryl 2-0). The intubation tube was removed just after the surgery. Four patients had no complication after the operation and one patient had temporary subcutaneous emphysema in the neck. For 4-11 months follow up after operation all the patients have had good breathing. Tracheal resection with end to end anastomosis is effective method of treatment of tracheal stenosis. PMID- 12894429 TI - [Anneroth, Batsakis and Luna's modified classification as prognostic factor in patients with laryngeal carcinoma]. AB - The cure rates for squamous cell carcinoma of the larynx have not improved over the past few decades. Despite the large number of therapeutic and histopathologic studies that have been performed, there are currently no clinical or morphologic markers available to predict outcome in patients with laryngeal carcinoma. The aim of the study was to exam tissues from 209 patients who were treated surgically because of advanced carcinoma of the larynx to create such a system. The examinations included routine histology and Anneroth, Batsakis and Luna's modified classification. The authors have analysed histologic parameters such as: cytoplasmic differentiation (kereatinization), nuclear polymorphism, number of mitoses per high-power field, mode and depth of invasion, plasmalymphocytic and eosinophil infiltration, to determine the potential usefulness of these factors in predicting recurrences and survival in patients with laryngeal cancer. PMID- 12894430 TI - [Levels of nonenzymatic antioxidants depending on stage of laryngeal cancer]. AB - The goal of this study was to check if there is any connection between nonenzymatic antioxidant system and larynx cancer staging. Levels of retinol, alpha- and gamma-tocopherol were measured in blood of 28 patients with larynx cancer. The conclusion is that the levels of measured substances were significantly lower in patients with more advanced larynx cancer. PMID- 12894431 TI - [Computerised tomography in pre-operative imaging of middle ear cholesteatoma]. AB - Computed tomography is today the best method of imaging to study chronic middle ear diseases. The aim of this study was to assess the diagnostic value of CT scan in cases of middle ear cholesteatoma in children. The preoperative CT scans were compared with the operative findings in retrospective analysis of 60 ears operated between 1998-2001. Our results show good radiosurgical correlation in cholesteatoma for most middle ear structures except for the integrity of long process of incus. The disadvantage of CT scans is inability to distinguish between cholesteatoma, granulation tissue and effusion. CT scans are an important investigative tool prior to cholesteatoma surgery. PMID- 12894432 TI - [Usefulness of TAPS (Polish version) for evaluation of CI children's progress in speech rehabilitation--7 years of observation]. AB - The speech rehabilitation of pre- and perilingually deafened children with cochlear implants it is a very long process. It should be individualized for needs of every young patient. It is possible only after correct evaluation of speech level. There are many tests for this evaluation. Among them same are more known like TAPS test. Test of Auditory Perception of Speech (TAPS) was elaborated in English at Basel University in Swiss. It was translated to Polish at Adam Mickiewicz University and Karol Marcinkowski University School of Medical Sciences in Poznan. The authors present their own 7-years observations. PMID- 12894433 TI - [Role of cytokines in pathomechanism of otitis media]. AB - Otitis media is one of the most common diseases in children. A variety if cytokines, potent inflammatory mediators, play a crucial role in the initiation and maintenance of the immunological response in otitis media. We discussed a profile of cytokines in various forms of otitis media (otitis media acute, chronic otitis media with effusion) and we underlined the differences between these types. We also described the influence of the most common pathogens of otitis media (S. pneumoniae and H. influenzae) on the cytokine panel in middle ear effusion. We discussed the current data supporting the hypothesis that in some cases the middle ear may act as a target organ for allergic inflammation. Since numerous authors demonstrated that the cytokine profiles are relevant for the evolution of otitis media it has been proposed that measurement of some proinflammatory cytokines in the middle ear effusion may be used in order to monitor the inflammatory process. It has been also proposed using cytokines as a target for pharmacological manipulation in the treatment of otitis media. PMID- 12894434 TI - [Influence of hypothermia and extracorporeal circulation on transiently evoked otoacoustic emission (TEOAE) in children operated on for various heart defects (I)]. AB - The aim of this study was to determine influence of extracorporeal circulation and body temperature on transiently evoked otoacoustic emission (TEOAE) in children operated on for various heart defects. The investigated group consisted of 44 children (average age 9.3 +/- 5.9 years). TEOAEs were measured one day before and 7-8 days following surgery. Statistical analysis revealed the influence of duration of extracorporeal circulation and depth of hypothermia on TEOAEs assessed following surgery. TEOAEs showed tendency to decrease in patients who were operated in long lasting extracorporeal circulation in normothermia. Such a tendency was not observed in patients operated in temperatures between 30 35 degrees C and extracorporeal circulation time between 1-2 hours. This observation confirms that hypothermia has a protective role for the cochlea and could prevent its damage during long lasting operations performed in extracorporeal circulation. PMID- 12894435 TI - [Prognostic factors in sudden deafness]. AB - Sudden sensorineural hearing loss is still a diagnostic and therapeutic dilemma and is very difficult to predict recovery in it. Different factors may influence a prognosis like severity of hearing loss, duration of symptoms before treatment, presence of vertigo, type of audiogram and age of patients. The aim of the study was an evaluation of the hearing improvement in sudden deafness in relation to some of these elements. A retrospective analysis of 119 patients treated for sudden sensorineural hearing loss during 10 years was done. Clinical otolaryngological, neurological and ophthalmological examination, audiologic and ENG tests were carried out. Hearing improvement was obtained in 51 patients (43%). Hearing recovery was recorded in 38 patients (66%) in whom duration of disease before treatment was no longer than 7 days, in 9 patients (25%) with a period 8-14 days and in 4 patients (16%)--with period 15-30 days (66% vs 25% vs 16%, p < 0.001, 25% vs 16% p = 0.39). Hearing improvement was more frequent in patients with initially mild (51.6%) than severe (38.7%) and profound hearing loss (25%) (51.5% vs 25%, p < 0.05). Hearing recovery was observed in 18 patients (33.3%) with vertigo and in 33 patients (50.8%) without vertigo (p = 0.056). In analysis of age (five groups: until 30 years, 31-40 years, 41-50 years, 51-60 years, over 60 years) in comparison with hearing recovery it was not found any significance (45.5% vs 45.8% vs 46.4% vs 37% vs 38.9%, p = 0.94). It was stated that in patients with sudden deafness duration of the disease before treatment and level of hearing loss may significantly influence an outcome, also vertigo may worsen a recovery, contrary age of the patients does not seem as important prognostic factor. PMID- 12894436 TI - [Physiotherapy of Eustachian tube dysfunction in adults--preliminary report]. AB - The paper presents preliminary results of Eustachian tube dysfunction treatment with the use of vibroaerosols in adult patients. 21 patients aged 16-73 y.o. (mean age 49 y.o) have been included in the study. The treatment included inhalation of medicines individually selected for the patient, based on the results of the physical examination. The patients have been divided into 2 groups, depending on the type of tympanogram. Group I included patients with type A tympanogram and abnormal Eustachian tube patency test result. Group II comprised of patients with type B or C tympanogram and abnormal Eustachian tube patency test result. After treatment, subjective improvement has been observed in 88.9% of the ears, objective improvement in 61.5% of the ears. In group II, objective improvement observed by the change of tympanogram has been observed in 85% of ears. Lack of correlation between objective and subjective improvement have been observed in 26% of the ears. In 11.1% of the ears no benefit from treatment has been observed. The correlation between subjective and objective improvement has been observed in 74% of ears. It appears that treatment with the use of vibroaerosols is beneficial for patients with Eustachian tube dysfunction. PMID- 12894437 TI - [Unusual case of the ear cholesteatoma]. AB - The authors present a case of temporal and occipital bone extensive destruction with epidural abscess of posterior cranial cavity and abscess of occipito-nuchal region created by giant cholesteatoma of the middle ear. There has been discussed a diagnosing troubles as well as performed surgical treatment. PMID- 12894438 TI - [Internal injuries of larynx in children. Case reports]. AB - The authors present larynx injuries among children especially with regard to iatrogenic internal injuries based on personally observed cases: dislocation of arytenoid cartilage in 11 year-old girl and post intubation granuloma in 8 year old girl. PMID- 12894439 TI - [Neuroendocrine carcinoma of the larynx--review of literature and report of 2 own cases]. AB - A review of the international medical literature was conducted regarding the laryngeal neuroendocrine tumors. This paper discusses the diagnosis, histopathology, treatment, and prognosis in laryngeal neuroendocrine tumors. It also discusses the heterogenous classification schema of such tumors. Two cases of a very rare neuroendocrine cancer of larynx are described. Both were localized in the supraglottic area and occurred in the 5th and 6th decades of life, consistent with previously reported cases of such neoplasms. PMID- 12894440 TI - [Katarrus, chrapota, slinogorz--names and treatment of laryngeal diseases on the basis of 15th and 16th century relics of Polish language]. AB - The paper outlines the beginnings of the process of naming laryngological diseases in the Polish language, and is based on 15th and 16th century relics of the Polish language. The authors relied in their analysis on the registers, as well as practical health guides such as herbarium books and strictly scientific medical manuals. Most of old Polish names of illnesses include original names based on the observation of the symptoms of illnesses and their clinical aspects. A great number of synonyms, both native and derived from foreign words, deserve special attention, indicating that attempts must have been made to find appropriate terms, which would best express the characteristics of a given illness. The presence of names derived from Latin and Greek should be attributed to the attempts at unifying medical terminology. Over the centuries, as the language and medical science have developed, names of illnesses have undergone a number of changes. It should be noted that numerous names derived from old Polish are still applied as medical terms. PMID- 12894441 TI - [Effect of glucocorticoids and hyperbaric oxygenation on the inner ear in clinical investigation of patients with sudden deafness and in experimental acoustic trauma in chicks]. PMID- 12894442 TI - [Assessment of the extent of angiogenesis and proliferation and expression of selected proteins connected with apoptosis in patients with laryngeal cancer]. PMID- 12894443 TI - [10th Danube Symposium International Otorhinolaryngologic Congress]. PMID- 12894444 TI - [12th Scientific Congress "Biomaterials in Medicine and Veterinary." October 20 23, 2002, Rytro, Poland]. PMID- 12894445 TI - [Report form the Polish Otorhinolaryngologic Congress--Head and Neck Surgery. Mikolajki, June 13, 2002]. PMID- 12894447 TI - The future of gene therapy. PMID- 12894448 TI - Angiogenesis clinical trials. PMID- 12894449 TI - AAV vectors, the future workhorse of human gene therapy. PMID- 12894450 TI - Nonviral vectors for cardiovascular gene delivery. PMID- 12894451 TI - Gene therapy for coronary artery disease: preclinical and initial clinical results with intracoronary administration of Ad5FGF-4. PMID- 12894452 TI - Cancer gene therapy: present status and future directions. PMID- 12894453 TI - Oncolytic virotherapy as a novel treatment platform for cancer. PMID- 12894454 TI - Adenovirus retargeting and systemic delivery. PMID- 12894455 TI - CNS diseases amenable to gene therapy. PMID- 12894456 TI - Molecular therapies for the nervous system and muscle. PMID- 12894457 TI - Lentivirus: a vector for nervous system applications. PMID- 12894458 TI - Endothelial progenitor cells for neovascularization. PMID- 12894459 TI - Therapeutic aptamers and antidotes: a novel approach to safer drug design. PMID- 12894460 TI - Plasmid-based gene transfer and antiprogestin-controllable transgene expression. PMID- 12894461 TI - [Testicular torsion can also be present in adult men]. AB - Five adult men, aged 34, 33, 31, 56 and 52 years respectively, were admitted with acute scrotal pain. Testicular torsion was eventually diagnosed in all the patients. Two of the patients were initially treated with antibiotics due to a suspected epididymitis. One of them required orchidectomy due to necrosis. The remaining patients underwent orchidofixation and their testicles could be preserved. The most important different diagnosis for acute pain and swelling of the testicle is either acute epididymitis or testicular torsion. The latter is uncommon in adult men. Using a physical examination to distinguish epididymitis from testicular torsion can be difficult. Doppler ultrasound is a reliable diagnostic tool for confirming testicular ischaemia. If the intratesticular flow is diminished or absent, then testicular torsion should be considered and this should be followed by immediate surgical exploration. PMID- 12894462 TI - [Neuromonitoring of patients following severe brain trauma]. AB - To a considerable extent, the neurological outcome of patients with severe brain trauma is determined by the primary injury caused by the accident. Substantial progress has been made in our understanding of the pathophysiological processes resulting in secondary brain damage after brain trauma, partly as a result of the introduction of advanced techniques of neuromonitoring. Early recognition and treatment of the symptoms leading to this type of brain damage seems crucial to the effort to improve the neurological outcome in these patients. Useful modern neuromonitoring techniques include: measurement of the intracranial and cerebral perfusion pressure and continuous electroencephalography. Methods that are also reliable and readily applicable, but less relevant clinically, include cerebral microdialysis of fluid from the extracellular space, determination of the parenchymal oxygen tension, and determination of the venous oxygen saturation. Other techniques that are not clinically applicable include: determination of the cerebral blood flow, the cortical tissue perfusion or the regional cerebral oxygenation. PMID- 12894463 TI - [Inhaled corticosteroids for COPD]. AB - Over 60% of patients with COPD are treated with inhaled corticosteroids (ICS), even though their use is still subject to debate. The inflammatory process in the lungs of patients with COPD is dominated by macrophages, CD8+ T-lymphocytes, neutrophilic granulocytes and mast cells, as well as an increased production and/or concentration of IL-8, TNF-alpha and leucotrine B4. This inflammatory process is less sensitive to the effect of corticosteroids than that in asthma. Placebo-controlled clinical studies show a beneficial effect of ICS on the number and severity of exacerbations, symptoms and quality of life, but not on the accelerated decline in lung function. Exacerbations occurred in a subgroup of patients in whom ICS were discontinued. A trial treatment with ICS should be considered in all COPD patients who have previously exhibited signs of asthma or allergy, or who have marked bronchial hyperresponsiveness, considerable reversibility, and recurrent exacerbations. In patients with moderate and severe COPD, a one-year trial with ICS should be considered, with exacerbations and symptoms as primary efficacy parameters and lung function decline as a secondary parameter. The combined use of ICS and long-acting beta-2 agonists may be considered in view of an additional effect on these parameters compared to administering the individual components. PMID- 12894464 TI - [Clinical reasoning and decision-making in practice. A young woman with fever, shortness of breath, and reduced consciousness]. AB - A 23-year-old woman with mild psychomotor retardation presented with fever, coughing, reduced consciousness and a stiff neck. A chest X-ray revealed an infiltrate in the left lower lobe; the cerebrospinal fluid was cloudy with a mild pleocytosis. Ceftriaxone was prescribed and the fever subsided. On the second day of admission she had a seizure, and a paraparesis emerged. Despite changes in the antibiotic regimen, her clinical condition hardly improved. On the fifth day, antibodies against Mycoplasma pneumoniae were found to be strongly positive and the diagnosis was M. pneumoniae infection. This accounted for the pneumonia together with meningoencephalitis and a transverse myelitis. The antibiotics were switched to doxycycline and the clinical condition improved dramatically. Six weeks after discharge, the patient had made a complete recovery. In patients suffering from meningitis with an atypical presentation, uncommon causes of infection should be considered. Together with a pneumonia, M. pneumoniae, Chlamydia pneumoniae, Legionella pneumophila and Listeria monocytogenes should be high on the list of potential causes for bacterial meningitis. PMID- 12894465 TI - [From gene to disease; hepatocystin and autosomal dominant polycystic liver disease]. AB - Polycystic liver disease (PCLD, MIM 174050) is a dominantly inherited condition characterised by the presence in the liver of multiple cysts of biliary epithelial origin. It must be distinguished from autosomal dominant polycystic kidney disease type 1 (ADPKD-1, MIM 173900) and type 2 (ADPKD-2). Both disorders may be complicated by polycystic liver disease, but renal involvement is absent in PCLD. PCLD is often asymptomatic, but if symptoms arise, they are usually due to the mass effect of cysts. The phenotype is more severe in females and correlates with the number of pregnancies or estrogen use. The gene for PCLD has been assigned to chromosome 19p13.2-13.1. Two separate large-scale positional cloning efforts have managed to identify PRKCSH as the gene underlying PCLD. Up to now, all mutations found in PRKCSH introduce stopcodons in the m-RNA, resulting in premature termination of translation to protein. This suggests a loss of function of the encoding protein. The protein, designated by us as hepatocystin, is predicted to be localised in the endoplasmic reticulum. Multiple biological roles have been suggested for hepatocystin, such as a substrate for phosphorylation by protein kinase C, binding to advanced glycation endproducts, and a function as the non-catalytic beta-subunit of glucosidase-II. The role of hepatocystin in PCLD, however, remains to be elucidated. PMID- 12894466 TI - [Diagnostic image (148). A man in coma after a traffic accident. Pneumocephaly]. AB - A 73-year-old man developed extended pneumocephaly due to a skull fracture after a severe traffic accident. PMID- 12894467 TI - [Estimate of the number of new patients with type 2 diabetes mellitus in the Netherlands: at least 65,000 per year in the age group of 50 years and above]. AB - OBJECTIVE: To estimate the incidence of type-2 diabetes mellitus in the Netherlands, using figures which were observed in a population-based cohort study of diabetes, the Hoorn Study. DESIGN: Descriptive. METHOD: Age-specific, gender specific and glucose-status-specific diabetes incidence figures as observed in the Hoorn Study (1989-1998) were applied to the Dutch population structure of January 1, 2000. RESULTS: Based on the fasting glucose diagnostic criteria, as recommended for clinical practice, 312,393 patients with incident type-2 diabetes were expected over 6.4 years in the 50-74 years age category, corresponding to an average of 48,811 patients per year. If data are extrapolated to the > or = 75 years age category, an additional 16,865 patients per year are expected (total: 65,676). If the 2-hour postload glucose values of the oral glucose tolerance test were also used to diagnose diabetes, these figures would be 66,001 new patients per year in the 50-74 years age group, and 22,840 in the > or = 75 years age group. PMID- 12894468 TI - [Invasive infections due to group A beta-haemolytic streptococci in two families]. AB - An invasive beta-haemolytic Lancefield group A streptococcal (GAS) infection was diagnosed in 4 patients: a 70-year-old woman, her 71-year-old husband, a 62-year old woman and her 43-year-old son. In the married couple the infection was caused by GAS-type TB3264M100. The woman had a pneumonia, whilst her husband developed a streptococcal toxic shock-like syndrome; he died. The other woman and her son were infected with GAS-type T6M6. The son died of a circulatory arrest due to necrotizing fascitis from a wound in his arm. His mother recovered following a severe tonsillitis. The number of invasive GAS infections has increased in the past decades. GAS infections occur mostly in isolated cases, but clusters of patients are also seen, like the two described here. The risk of an invasive GAS infection is greatest if one has been in the neighbourhood of the index patient during the week prior to the diagnosis in that patient. According to the latest (American) guidelines, there is no reason for prophylactic treatment of the close contacts of patients. PMID- 12894469 TI - [Intensive care medicine in the Netherlands, 1997-2001]. PMID- 12894470 TI - [Favorable results of conservative treatment with isosorbide dinitrate in 25 patients with fourth-degree hemorrhoids: a pilot study]. PMID- 12894471 TI - [The physician as an expert in medical negligence cases]. PMID- 12894472 TI - [Interspecies transmission of influenza A viruses and influenza pandemics]. AB - Molecular and genetic data are summarized on the origin of influenza A virus pandemic variants. Conceptual modifications of the reassortment theory of the origin of pandemic strains are discussed in connection with the appearance of new H5 and H9 avian influenza viruses, which caused the respiratory infection in man and which are presently in the focus of attention as possible agents of future pandemic. PMID- 12894473 TI - [Antigenic and genetic properties of Seoul virus strains--a causative agent of hemorrhagic fever with renal syndrome]. AB - Data on the circulation of a new genetic variant of Seol virus, which was designated as "Vladivostok" (the name of the place where it was isolated)--VDV, were obtained, by using the antigenic and genetic analysis of hantavirus strains isolated from patients with hemorrhagic fever with renal syndrome (HFRS) and from brown rats, and described in the present paper. A complete identity of nucleotide sequences of the M-segment fragment of isolates obtained from HFRS patients and of those obtained from rats was demonstrated, which confirms the key role of brown rat as a source of infection. PMID- 12894474 TI - [Changes of humoral and cellular immunity in chronic hepatitis C patients of different staging]. AB - The purpose of the present study was to investigate the influence produced by viral proteins in the hepatic cells and RNA of hepatitis C virus (HCV) on the indices of T- and B-cell response in 52 patients with chronic hepatitis C (CHC). A relative count of peripheral-blood lymphocytes (PBL), expressing antigens CD3+, CD4+, CD8+, CD16+, CD20+ and CD95+ was estimated. The repertoire of antibodies to HCV proteins was specified. The thus obtained data were compared with an activity and a disease stage by using the histological diagnosis and alanine-amino transferase (ALT) level as well as with the presence of HCV RNA in the serum and viral protein of the liver. Such comparison of data and the use of the correlation analysis made it possible to establish that the antibodies to NS5 protein were detected reliably more often in patients with a more pronounced hepatic fibrosis, with a higher ALT activity and with expression of HCV proteins in the liver. At the same time, the presence of proteins in the liver and of RNA in the serum were accompanied by a more active humoral response to the non structure proteins of NS4 and NS5 as well as by more profound discrepancies of the immunity T-cell chain (a lowered ratio of CD4+/CD8+ and a smaller content of CD95+). There were no differences between PBL of the studied populations in patients with various activities and an HCV stage. A relatively bigger quantity of CD95(+)--positive PBL was found to be reliably higher in patients with viremia but lower in those cases, in which HCV proteins were detected in the liver. This confirms the inhibiting ability of HCV proteins to the Fas-mediated apoptose of PBL in CHC patient. PMID- 12894475 TI - [Antibodies to herpesvirus type 8 in Kaposi's sarcoma patients and controls in Russia]. AB - The purpose of the present study was to investigate the HH-8 seroprevalence among patients with Kaposi's sarcoma (KS), melanoma and gastric carcinoma (GC) as well as among renal recipients and blood donors. The obtained data revealed a high percentage of seropositive KS patients, which ranged from 83.6% in the classical disease type to 68.8% and to 71.4% in the immunosuppressive and AIDS-associated disease types, respectively. On the whole, the positive humoral response to HHV-8 reliably correlated with the positive findings if the viral genetic information in tumor tissue samplings. An unexpectedly high percentage of seropositive persons was found among the GC patients (41.8%) and among the renal recipients (26%), which is apparently predetermined by the immunosuppressive condition of such patients. Seroprevalence was found only in 4% of blood donors. Thus, the obtained data make it possible to conclude that KS cases, as diagnosed in Russia, are tensely associated with HHV-8 in spite of a low virus spread among the healthy population. Patients with pathology concomitant with a pronounced immunosuppression are characterized by a high prevalence of HHV-8 and belong to the category of persons with a KS risk. PMID- 12894476 TI - [Specific determination of flaviviruses by molecular hybridization with synthetic deoxyoligonucleotide probes]. AB - Molecular probes were designed for the purpose of specific determination of flavioviruses transmitted by the ticks of tick-borne encephalitis (TBE) and Omsk hemorrhagic fever (OHV) as well as by mosquitoes of Japanese encephalitis (JE), North Nile (NN), Murrey Valley encephalitis (MVE), Saint-Lois encephalitis (SLE), dengue 1-4 and of yellow fever (YF). The probes are synthetic deoxyoligonucleotides with the 18-20 long basis and complementary for the RNA fragments defined by computer analysis. The thus obtained probes, which specifically hybridize themselves with the sets of the TBE virus or of the OHV virus and do not hybridize themselves with other TBE viruses' sets. Group specific probes for YE and dengue viruses as well as virus-specific probes, which are able to detect each of the above viruses without any cross effects, were suggested for indexing and identifying the flaviviruses transmitted by mosquitoes. PMID- 12894477 TI - [Differences in oligomerization of nucleocapsid protein of epidemic human influenza A(H1N1), A(H1N2) and B viruses]. AB - A comparative analysis of involving the nucleocapsid protein (NP) into shaping-up of SDS-resistant oligomers was carried out presently in circulating epidemic strains of human influenza, viruses A and B. The study results of viral isolates obtained from clinical samples and recent standard strains revealed that the involvement of NP in the SDS-resistant oligomers, which are different in various subtypes of influenza A viruses. According to this sign, the human viruses A(9H3N2) are close to the avian ones, in which, as proved by us previously, virtually the entire NP transforms itself into the oligomers resistant to SDS. About 10-20% of NP are involved in shaping-up the virus influenza A(H1N1) of SDS resistant oligomers. No SDS-resistant NP-oligomers were detected in influenza of type B. It is suggested that the prevalence of human viruses A(H3N2) in NP oligomers are the peculiarities of NP structure and of the presence of the PB1 protein from avian influenza virus. PMID- 12894478 TI - [Detection of type specific antibodies to hepatitis C virus NS4 protein in hepatitis C patients by enzyme immunoassay]. AB - A multi-enzyme immmune-assay test system was designed for serotyping of genotypes hepatitis C virus (HCV) and a method of such typing of the serum of patients with hepatitis C was worked out. The above test-system was worked out on the basis of a study of 10 type-specific peptides modeling different fragments from NS4 protein variable region of HCV. The designed test system was evaluated by using a set of 42 serum samples obtained at random from patients with chronic hepatitis C, which had been preliminarily genotyped by polymerase chain reaction. The serotyping makes it possible to identify the type-specific antibodies in the blood sera of patients, including those cases when viremia was absent. Differences in the circulation of HCV in Moscow (Russia) and Vitebsk (Byelorussia) were established by using the designed test-system. PMID- 12894479 TI - [Expression of hepatitis A virus procapsids in the insect cells infected by recombinant baculovirus]. AB - The recombinant baculocvirus containing genome P1-2A-P3 of hepatitis A virus (HAV) was constructed and used for infecting the Sf9 insect cells. It was demonstrated that the deletion of 2BC from HAV polyprotein and the insertion of a new 3C protease cleavage site between P1-2A and P3 did not interfere with the processing of polyprotein or with forming the 70S-procapsids. The identity of the protein contents as well as of morphological and antigen characteristics, obtained in Sf9-cells, to HAV empty capsids, which take shape in the infected mammal cells, proves that it is possible to use them in making the vaccine and diagnostic preparations. PMID- 12894480 TI - [Clinical and laboratory evaluation of efficiency and tolerance of the "Nikavir" in acute hepatitis B]. AB - New antiviral drug "Nikavir" was clinically tested in acute hepatitis B (AHB). A total of 60 AHB patients were followed up; 30 patients were in the main group and 30 persons were in the control group. The treatment with Nikavir of AHB patients was shown to ensure a favorable clinical disease course and it was accompanied by a quicker rate in reduction of the activity of serum enzymes AIAT, AcAT, alkaline phosphatase and of gamma-glutamil-transpeptidase; besides, it was accompanied by an early elimination of such serum markers of AHB like HBeAg, HbsAg and HBV DNA. Wikavir can be used in the treatment of patients with mild and moderate forms of AHB. PMID- 12894481 TI - [Development of "Amplisens-HCV-genotype" reagent set for identification of hepatitis C virus genotypes 1a, 1b, 2a and 3a]. AB - Multiple alignments of 119 nucleotide sequences of isolates of hepatitis C virus (HCV) were carried out to choose the type-specific primers for the 5'-ultra-core fragment of viral genome for the purpose of detecting the HCV 1a, 1b, 2a, and 3a subtypes. A PCR kit of reagents was designed for the amplification of cDNA HCV with selected type-specific primers and for making the electrophoresis in agarous gel. The kit comprises the positive control samples, i.e. HCV genome fragments, subtypes 1a, 1b, 2a and 3a, cloned in the plasmid vector. 440 cDNAHCV samples were simultaneously tested by using the worked out reagents' set and according to the method of Ohno et al. The results were found to be concordant in 336 cases, and were discordant in 4 samples. A sequencing of the PCR products and phylogenetic analysis showed that 1 sample belonged to subtype 4a, 2 samples belonged to subtypes 2k and 1 sample--to subtype 31. PMID- 12894482 TI - Clinical importance of the genus Trichoderma. A review. AB - Opportunistic fungal infections have been observed with increasing frequency in recent years in immunocompromised patients. Several data were published in the last decade about the clinical importance of the filamentous fungal genus Trichoderma, indicating that Trichoderma strains--besides their agricultural and biotechnological importance--may be potential opportunistic pathogens in immunocompromised hosts as well. This review is going to summarize the clinical case reports about Trichoderma infections, and to discuss the information available on the antifungal susceptibility and on the ecophysiological, enzymological and systematic aspects of clinical Trichoderma isolates. PMID- 12894483 TI - In vitro activity of cefditoren against a special collection of clinical isolates of Streptococcus pneumoniae from Hungary. AB - Cefditoren is the active form of cefditoren pivoxil, a new, broad-spectrum oral cephalosporin with strong in vitro activity against penicillin-susceptible and resistant Streptococcus pneumoniae. In this study, the minimum inhibitory concentrations (MICs) of cefditoren were determined for a special selection of S. pneumoniae isolates known to be susceptible, moderately susceptible or fully resistant to penicillin; these isolates originated from the lower respiratory tract of adults with pneumonia or the upper respiratory tract of children with or without symptoms of infection. Some of this latter group of isolates exhibited extremely high MICs to penicillin (> or = 32 mg/l), whereas the MICs of cefditoren did not exceed 2 mg/l. The MIC50 and MIC90 of cefditoren proved to be 0.25 and 1.0 mg/l, respectively, with a range of MICs < or = 0.015-2.0 mg/l for all the tested S. pneumoniae isolates. Its good activity suggests that cefditoren is expected to be a potent drug in infections caused by penicillin-resistant and multidrug-resistant S. pneumoniae. PMID- 12894484 TI - Regulation of Trichoderma cellulase formation: lessons in molecular biology from an industrial fungus. A review. AB - The present article reviews the current understanding of regulation of cellulase gene transcription in Hypocrea jecorina (= Trichoderma reesei). Special emphasis is put on the mechanism of action of low molecular weight inducers of cellulase formation, the presence and role of recently identified transactivating proteins (Ace1, Ace2, Hap2/3/5), and the role of the carbon catabolite repressor Cre1. We also report on some recent genomic approaches towards understanding how cellulase inducers signal their presence to the transcriptional apparatus. PMID- 12894485 TI - [ Systemic and cardiac B Fibrillosis (" Amyloidosis"). A clinical comment]. PMID- 12894486 TI - [Participation of nitric oxide and arachidonic acid metabolites via cytochrome - P450 in the regulation of arterial blood pressure]. AB - Nitric oxide and cytochrome P450 arachidonic acid metabolites participate in blood pressure regulation. The synthesis of these autacoids leads to arterial hypertension. However, it is not known whether there is an interaction between them. Therefore, we studied the modulatory effect of nitric oxide and cytochrome P450-arachidonic acid metabolites, their interaction on blood pressure, and the renal content of cytochrome P450. Male Wistar rats were divided: 1) control, 2) L NAME (100 mg/kg/d p.o.), 3) L-NAME + SnCl2 (10 mg/kg/d i.p.), and 4) L-NAME + dexamethasone (1 mg/kg/d s.c.). We measured blood pressure and collected urine and blood for nitric oxide measurement. NO2 was quantified by HPLC. Blood pressure was: control, 97 +/- 7 mmHg; L-NAME, 151 +/- 4.6 mmHg; L-NAME + SnCl2, 133 +/- 3 mmHg, and L-NAME + dexamethasone 152 +/- 4.5 mmHg. Urine nitrite concentration was: 1) 1.832 +/- 0.32, 2) 1.031 +/- 0.23, 3) 1.616 +/- 0.33, and 4) 1.244 +/- 0.33 mumol/mL, while the concentration in blood was: 1) 0.293 +/- 0.06, 2) 0.150 +/- 0.05, 3) 0.373 +/- 0.13, and 4) 0.373 +/- 0.07 mumol/mL. L NAME + SnCl2 decreased cytochrome P450 renal content, and L-NAME + dexamethasone showed a similar response. In conclusion, both, nitric oxide and CYP-arachidonic acid metabolites play a role in the regulation of blood pressure. Nitric oxide also partially regulates renal cytochrome P450 content. PMID- 12894487 TI - [Coronary heart disease in Mexico and the clinical epidemiological and preventive relevance]. AB - Coronary heart disease is dominant among heart diseases in the population, a problem to control. Heart diseases have been first place in the general mortality for the last 20 years. The trend show an annual increase without control. Atherosclerosis is responsible for at least for one fourth of all deaths in the country. The combined data of mortality and morbidity, from hospital discharges or the whole country, disclose that one out of three die of acute myocardial infarction (AMI). While only 8% of all deaths registered were admitted to a hospital. Most, 92% were never admitted to any hospital for some reason. The estimated annual incidence of AMI cases is 140,000, on the basis of 3 cases surviving for each case death, accounted by the Death Certification System, which rounds 35,000. Standardized mortality rates of AMI in Mexico are greater than in USA o Canada creating a more concerned worry. The most probable explanation to that situation is lack of preventive care, which should also include the acute care and before reaching the hospital facilities. Running the same chances are hypertension crisis and strokes and diabetic complications. The appropriate care for critical situations might reduce significantly the cardiovascular mortality in the country, in a short and middle term. Prevention is not only for chronic conditions but for acute and critical situations. The programs of preventive care should also include cultural promotion and community awareness. The timely care is life and myocardium saving. The reinforcement of prior to hospital care reduces the delay for AMI adequate intervention. These activities agree quite well with the ongoing programs of CPR, organizing the surviving chain and the training programs for paramedical emergency technicians. PMID- 12894488 TI - [Echocardiographic diagnosis of anomalous origin of one pulmonary artery from the ascending aorta]. AB - OBJECTIVE: To present our experience in the echocardiographic diagnosis of anomalous origin of one pulmonary artery from the ascending aorta (AOPA). To analyze its clinical presentation, treatment and outcomes with special emphasis in the echocardiographic data. METHOD: We reviewed restrospectively the clinical, hemodynamic data, and surgical findings of patients with an echocardiographic diagnosis of AOPA studied in the Hospital Infantil of Mexico "Federico Gomez" from 1991 to 2002. RESULTS: The study includes 12 children with AOPA; Seven were males. The average age at diagnosis was two months; 4 in neonatal period, 3 under 1 year and 5 older than 1 year. The diagnosis was established prospectively by echocardiography in all patients and it was confirmed by angiography in 8 and at surgery in 9. Ten had anomalous origin of right pulmonary artery. The associated anomalies were patent ductus arteriosus in 6, ventricular septal defect in 2 and aortopulmonary window, atrioventricular discordance, double outlet right ventricle and tetralogy of Fallot in one case each. Nine underwent corrective surgery of all the anomalies. One patient died on the sixth postoperative day; the remaining are in good condition without stenosis at the site of the anastomosis. Surgery was refused in one. One patient was not candidate to surgery due to advanced obstructive pulmonary vascular disease (OPVD) and one case is awaiting surgery. CONCLUSION: The diagnosis of AOPA may be established with precision through echocardiography. Cardiac catheterization is seldom needed to confirm morphology but is mandatory in older children with suspected of OPVD. PMID- 12894489 TI - [ Segmental ostial ablation to interrupt electrical conduction in a single pulmonary vein for the prevention of idiopathic paroxysmal atrial fibrillation]. AB - Several studies have shown the role of focal trigers in the pulmonary veins initiating episodes of atrial fibrillation. Radiofrequency catheter ablation of this foci is a curative therapy for paroxysmal atrial fibrillation. We report a case of idiopathic paroxysmal atrial fibrillation triggered by abnormal electrical activity in a single pulmonary vein. Mapping was performed during sinus rhythm with a 4F decapolar catheter (Spiral Supreme, Daig, St. Jude Medical) positioned near the ostium by a transseptal approach. Pulmonary vein potentials were only identified in the left superior pulmonary vein. Segmental ostial ablation (30 W) performed during left atrial pacing resulted in complete cesation of conduction in the pulmonary vein. There were no complications. The clinical response (suppression of the paroxysms of atrial fibrillation in a 9 month follow-up) observed in this patient imply that atrial fibrillation was triggered by this pulmonary vein. This case report illustrates several aspects of catheter-based ablation of pulmonary vein foci for the treatment of paroxysmal atrial fibrillation and demonstrates its efficacy. PMID- 12894490 TI - [Apical hypertrophic cardiomyopathy without aneurysm. Diagnosis trough myocardial gammagraphy and magnetic resonance]. AB - We present three patients with apical hypertrophic cardiomyopathy, their diagnosis was performed by means of imaging techniques to determine morphology, severity, configuration, complications, and distribution of the ventricular hypertrophy. Through color-flow and pulsed Doppler, the protosystolic flow in the midventricular region was detected, which we believe corresponds to the obstruction due to coarctation of the walls during early ventricular systole and that this gradient is independent from the one identified as paradoxical in patients with apical aneurysm. Explorations by means of magnetic resonance and Gated-Spect myocardial gammagraphy revealed the apical location of the hypertrophy confirming the "spadelike" configuration of the left ventricle and the absence of apical aneurysm. PMID- 12894491 TI - [Electrical features of hypertrophied left heart]. AB - Chronological and spatial characteristics of the main resultant vectors of the left atrial and ventricular depolarization in normal conditions and in presence of hypertrophy, due to a sustained overload, are described. The coexistence of interatrial, intraatrial, and intraventricular conduction disorders can modify the orientation of these vectors. The main electrocardiographic sign of left atrial hypertrophy is a P wave duration > 0.10 sec in adults. In case of left ventricle hypertrophy, the time of onset of the intrinsicoid deflection (TOID) is prolonged in the near left unipolar leads, and the S wave voltage is increased in opposite regions, i.e. in the right precordial or transitional leads. It is necessary to bear in mind other useful electrocardiographic signs. Hence, absence of the right basal vector (IIId) manifestation in a clockwise rotating heart is probably due to an increase in the basal electromotive forces of the left ventricle (vector IIIi) due to hypertrophy or ipsilateral ventricular conduction disorders. For a correct evaluation of these signs, it is mandatory to perform a rational analysis of the traings, not just a stereotyped electrical exploration. Besides it is very important to determine the Q-Tc interval in the left unipolar leads to establish whether, in these leads, the inverted T wave is of secondary type (normal Q-Tc) or of primary type (prolonged Q-Tc) due to a coexisting subepicardial or transmural ischemia. From these considerations, the usefulness of the thoracic circle and high abdominal unipolar leads is inferred. PMID- 12894492 TI - [Atrioventricular nodal reentrant tachycardia: Radiofrequency ablation]. AB - Atrioventricular nodal reentrant tachycardia is the most frequent regular supraventricular tachycardia. It is generally a benign arrhythmia and usually it is no associated with heart disease. It is more often seen in middle-aged women but it is no infrequent in younger and older patients. The development of radiofrequency with catheter ablation has provided the possibility to definitely cure the arrhythmia and moreover permit a better comprehension of the underlying electrophysiologic and anatomic substrate. PMID- 12894493 TI - [Calcified aneurysm of the left ventricle]. PMID- 12894494 TI - Molecular heterogeneity in human osteosarcoma demonstrated by enriched mRNAs isolated by directional tag PCR subtraction cloning. AB - Directional tag PCR subtractive hybridization was applied to construct a cDNA library generated from three different human osteosarcoma (OS) target cell lines (OHS, SaOS-2 and KPDXM) from which normal osteoblast (NO) sequences were subtracted. After two consecutive subtractive steps more than 98% of the common mRNAs species were depleted, leading to effective enrichment of the remaining target sequences. After differential screening of 960 clones, 81 candidates were further studied by Northern blot analysis and 73 represented separate mRNA species. Fifty-three of these showed enriched mRNA levels, of which 36 represented known and 17 not previously published cDNAs or EST sequences. The mRNAs showed a 1.4- to 504-fold enrichment compared to the mRNA levels in NO cells. The known mRNAs are: Ribosomal protein S11, KSP-37, Tethering factor SEC34, FXYD6, Alpha enolase, G-s-alpha, GPR85, DAF, RPL35A, GIF, TAPA-1, ANAPC11, DCI, hsp27, MRPS7 homolog, eIF p110 subunit, DPH2L, HMG-14, FB1 protein, chondroitin-6-sulphonase, calgizzarin, RNA polymerase II subunit, RPL13A, DHS, gp96, HHP2, acidic ribosomal phosphoprotein P2, ANT-2, ARF1, AFG3L2, SKD3, phosphoglucoisomerase, GST pi, CKI gamma 2, DNA polymerase delta small subunit and TRAP delta. Sections of human osteosarcoma biopsies and a xenograft were studied by in situ analysis. Seven cDNAs highly expressed in Northern blot analysis were tested. Their in situ expression differed between the xenograft and human sections as did that of collagen I. In the xenograft made from one of the target cell lines (OHS), a fair to strong representation of 3 cloned mRNAs was observed while collagen I mRNA was not detectable. We conclude that the molecular heterogeneity of these tumors is considerable. These results ought to have implications for future work to describe phenotypic subtypes with the aim of improving the diagnosis of human osteosarcomas. PMID- 12894495 TI - Cyclin D1 expression in the intestinal mucosa and tumors of Apc1638N mice. AB - Altered expression of cyclin D1 contributes to the development of several types of cancer, including colorectal cancer. This study examined cyclin D1 expression in 32 intestinal tumors in different stages of tumorigenesis in Apc1638N mice, a mouse model for human familial adenomatous polyposis (FAP). Three morphological patterns of expression of cyclin D1 in intestinal epithelial cells were found: nuclear, punctate-cytoplasmic and fine granular cytoplasmic. The nuclear pattern of cyclin D1 was detected in all of the tumors, including adenomas (n = 18) and adenocarcinomas (n = 14); this pattern was found predominantly in the tubular region of the tumors and in flat mucosa adjacent to a subset of the tumors (67% of adenomas and 57% of carcinomas). The punctate-cytoplasmic pattern of cyclin D1 expression was found in all adenocarcinomas and a majority of adenomas (80%), mainly in invasive and villous areas of the tumors; it was not found in normal flat adjacent mucosa suggesting that this pattern and altered cytoplasmic/nuclear expression were associated with tumor progression. Fine cytoplasmic granules were located in normal duodenum in the basal portion of the crypts and in colon in epithelial cells at the surface of the colonic crypts; in both duodenum and colon the number of cells with fine cytoplasmic granules significantly increased after feeding a Western-style diet. These altered patterns of expression of cyclin D1 may provide useful biomarkers of abnormal cell development for studies of tumorigenesis and the effects of chemopreventive agents. PMID- 12894496 TI - In situ RNA-RNA hybridisation of phospholipase C beta 3 shows lack of expression in neuroendocrine tumours. AB - BACKGROUND: Phospholipase C beta 3 (PLCB3) plays an important role in the signal transduction of the seven transmembrane receptors. The gene is located in the vicinity of the Multiple Endocrine Neoplasia type 1 (MEN1) gene on chromosome 11q13. Transfection of PLCB3 to neuroendocrine cell lines lacking expression suppresses the neoplastic phenotype and affects the gene expression of S100A3 and human mismatch repair protein, suggesting a role for PLCB3 in neuroendocrine tumorigenesis. MATERIALS AND METHODS: We used RNA-RNA in situ hybridisation for PLCB3 on a total of 82 samples including 34 from MEN1 patients. RESULTS: We show that the PLCB3 transcript is missing in 8 out of 14 MEN1-associated neoplasias as well as in 4 out of 10 bronchial carcinoids, 2 out of 10 exocrine pancreatic cancers and one sporadic adrenocortical carcinoma. CONCLUSION: Low or lack of PLCB3 expression in a subset of endocrine tumours, together with earlier published in vitro data on suppressor characteristics upon transfection, indicate that PLCB3 could be involved in the tumorigenesis in a subset of endocrine tumours. PMID- 12894497 TI - Expression of MMP-2, -9 and -13 in cell lines and fresh biopsies of squamous cell carcinomas of the upper aerodigestive tract. AB - BACKGROUND: The expression patterns of the matrix metalloproteinases (MMP) 2, 9 and 13 have been examined in biopsies and cell lines of squamous cell carcinomas (SCC) of the upper aerodigestive tract to determine the association between MMP expression and the aggressiveness of the tumor. MATERIALS AND METHODS: The expression of MMP-2, -9 and -13 was analyzed by immuno-hybridization in 12 keratinocyte cell lines, 13 SCC cell lines and in 63 biopsies from SCCs of the upper aerodigestive tract. RESULTS: In biopsies a correlation was detected between T-status and the MMP expression [MMP-2 (p < 0.05), MMP-9 (p < 0.01), MMP 13 (p < 0.01)], between N-status and MMP expression [MMP-2 (p < 0.05), MMP-9 (p < 0.01)] and between UICC stage and MMP-9 expression (p < 0.01). CONCLUSION: The MMP-2, MMP-9 and MMP-13 expressions correlate strongly with pathological parameters. The determination of these tumor-biological parameters could help to identify high risk patients with increased frequency of metastatic spread. PMID- 12894498 TI - Her2/neu expression in C-cell hyperplasia and medullary thyroid carcinomas. AB - Medullary thyroid carcinomas (MTC) are rare neoplasms derived from the calcitonin producing thyroid parafollicular C-cells. Interestingly about 20% of cases are related to inherited tumor syndromes. As precursor lesion, C-cell hyperplasia can be detected in numerous MTCs. In our series of 6 patients with MTC, including 5 patients with adjacent C-cell hyperplasia, Her2/neu levels were immunohistochemically evaluated on formalin-fixed, paraffin-embedded tissues using c-erbB-2/Her-2/neu Oncoprotein Ab-17 monoclonal antibody(mAb). Statistically, a highly significant correlation was found between high Her2/neu levels and extrathyroidal growth. C-cell hyperplasias always presented with higher amounts of stained cells than their corresponding invasive MTCs. Whereas in hyperplastic areas large stained cell clusters were found, the invasive tumors showed mainly single cells reacting in areas of variable size. These results present the impact of Her2/neu oncoprotein concerning development and biological behaviour, especially aggressive growth, of invasive MTCs and suggest a major role of hyperplastic C-cell areas in the development of these malignant tumors. PMID- 12894499 TI - Use of a SCID mouse model to select for a more aggressive strain of prostate cancer. AB - BACKGROUND: Prostate cancer is the most common non-cutaneous malignancy to affect men and has a propensity for metastasizing to bone. To better mimic the biology of metastatic prostate cancer, we have developed a model that utilizes both human prostate cancer and human bone in the SCID mouse. MATERIALS AND METHODS: Injection of a xenograft of human prostate cancer, LAPC-4, near a human bone core that was previously implanted within the hindlimb of a SCID mouse allowed for the selection of a more aggressive subset of cells, known as LAPC-4(2) (read as "LAPC 4 squared"). RESULTS: As compared to LAPC-4 cells, these "bone-selected" LAPC 4(2) cells form tumors more rapidly, develop PSA-positive serum at an earlier time-point as well as with higher levels, develop androgen independence, and metastasize to human bone after orthotopic injection. CONCLUSION: The selection of a more aggressive subset of prostate cancer cells that have developed androgen independence and the propensity to metastasize is of paramount importance as these are the cellular characteristics that are clinically linked to morbidity. Analysis of these "bone-selected" cells may lead to a better understanding of the molecular basis behind the conversion of low-grade prostate carcinoma to its more deadly, metastatic form. PMID- 12894500 TI - Analysis of signal transducer and activator of transcription 3 (STAT3) in gastrointestinal stromal tumors. AB - Several signaling pathways have been recognized in normal c-kit-mediated signal transduction following stem cell factor (SCF) stimulation including Janus kinase (JAK)/signal transducer and activator of transcription (STAT), mitogen-activated protein kinase (MAPK) and phosphoinositol 3-kinase (PI-3 K) pathways. In gastrointestinal stromal tumor (GIST), c-kit activation is considered to play a central role in its tumorigenesis. However, the signal transduction cascades specific for the SCF-independent c-kit activation in GIST remains to be elucidated. In this study, we examined for the expression of the activated form of STAT3 [phospho-STAT3 (tyr 705)] in eleven cases of GIST by immunohistochemistry. All GISTs had strong nuclear and variable cytoplasmic expression of phospho-STAT3 (tyr 705). Survival and proliferation of two established primary GIST cell lines with c-kit exon-11 mutations were then assessed for their response to inhibitors of c-kit (STI-571), JAK 2 (Tyrphostin AG490), MAPK kinase (PD98059) and PI-3 K(LY294002). GIST cells showed significant inhibition of proliferation and apoptosis when treated with STI571 or AG490 but not in cells treated with PD98059 or LY294002. Bcl-2 was expressed in all of the GIST cases (11 out of 11) and was down-regulated in the primary GIST cells following treatment with AG490. This study demonstrates that STAT3 is constitutively activated in GIST and JAK2 blockade leads to tumor growth inhibition and apoptosis indicating the involvement of the JAK/STAT signaling pathway in GIST cellular survival. PMID- 12894501 TI - Non-dysplastic and dysplastic Barrett's mucosa and adenocarcinoma lack a subepithelial myofibroblastic cell layer. AB - A subepitheal myofibroblastic (SMF) cell layer has been described in the colon, and referred to as pericryptal myofibroblastic cell layer. SMF cells have been shown to produce basement membrane proteins, including type IV collagen and laminin. The aim of this work was to determine the status of the SMF cell layer in Barrett's metaplasia (BM), with and without dysplasia, and compare that to the previously reported distribution of SMF in normal colon and colonic adenomas and carcinoma. Sections of formalin-fixed, paraffin-embedded biopsies from 6 colonic adenomas and 5 colonic adenocarcinomas, as well as 4 cases of BM without dysplasia, 4 with low grade dysplasia, 4 high grade dysplasia and 4 with invasive adenocarcinoma were immunohistochemically stained for alpha smooth muscle actin using the immunoperoxidase method. A continuous layer of SMF cells was present in all normal colonic tissue and adenomas but was absent in all colorectal adenocarcinomas. Surprisingly, none of the cases of BM with or without dysplasia or carcinoma showed an organized SMF cell layer. Unlike the colon, the SMF cell layer is absent in BM even without dysplasia. We hypothesize that lack of the SMF cell layer in BM may contribute to the quick progression to esophageal adenocarcinoma, which, unlike that in the colon, occurs before an exophytic lesion becomes evident. PMID- 12894502 TI - Diverse responses between human pancreatic cancer cell lines to native alpha 1 antitrypsin and its C-terminal fragment. AB - BACKGROUND: Previous studies imply that human pancreatic cancer cells have a wide heterogeneity and their exposure to various agents may give unpredictable results in clinical situations. MATERIALS AND METHODS: The cell lines LPC-3, -5 and -10, established from primary cultures of pancreatic adenocarcinoma, were exposed to 5 microM of AAT or its C-terminal peptide C-36 for 24 hours and analysed for cytokines by an enzyme-linked immunosorbent assay and for NF kappa B by the electrophoretic mobility shift assay. RESULTS: Native AAT lowers TGF-beta 1 levels and increases NF-kappa B activity in LPC-3 cells, while C-36 increases TGF beta 1 levels and up-regulates NF-kappa B in LPC-5 cells. In LPC-10 cells AAT lowers TGF-beta 1. However, both AAT and C-36 fail to cause a change in NF-kappa B expression. For LPC-10 cells treated with C-36 IL-6 and TNF-alpha levels also increase. CONCLUSION: Our findings provide evidence that human cancer cell lines originating from primary pancreatic tumors do not have a uniform response to the same stimulus which shows a great heterogenicity among pancreatic cancer cells. Serine proteinase inhibitor, AAT, dependent on its molecular form, is also found to exert diverse effects on the properties of tumour cells confirming the complexity of cell-protein interaction. PMID- 12894503 TI - Inhibition of cisplatin-induced ATR activity and enhanced sensitivity to cisplatin. AB - Ataxia teleangiectasia mutated (ATM) kinase, ATM-Rad3-related (ATR) kinase and DNA-protein kinase (DNA-PK) belong to a subgroup of protein kinases which play a role in the DNA damage response. In this study, cisplatin was shown to increase ATR activity and decrease ATM and DNA-PK activity. Caffeine, a nonspecific inhibitor of ATR, enhanced the cytotoxic effect of cisplatin, modestly decreased the p53 and p21WAF-1 response to cisplatin, and affected the cdc2-p34/cyclin B1 complex by decreasing both cyclin B1 protein accumulation and cdc2-p34 tyrosine 15 phosphorylation. The observed alteration of several potential ATR downstream targets suggests that inhibition of ATR activity may be one of the mechanism by which caffeine regulates sensitivity to cisplatin. PMID- 12894504 TI - Genomic alterations in human mesothelioma including high resolution mapping of common regions of DNA loss in chromosome arm 6q. AB - BACKGROUND: Molecular genetic analysis of 14 freshly resected human mesotheliomas was used to identify regions in the tumor genomes that display DNA copy number alterations, especially the regions that may harbor tumor suppressor genes. MATERIALS AND METHODS: Three methods for molecular analysis were used, comparative genomic hybridization (CGH), loss of heterozygosity (LOH) and a new method, quantitative microsatellite analysis (QuMA). RESULTS: The most frequent alteration detected by CGH and LOH was deletion in chromosome 6q, for which QuMA was performed at 30 different loci so as to define the region(s) of common deletion(s). Our data indicates that there are three independent regions of common deletion, one of size 8.4 Mb located at 6q14, a second of size 15.9 Mb at 6q22 and a third of size 12.0 Mb at 6q24. CONCLUSION: This suggests that at least 3 tumor suppressor genes mapped to chromosome 6q are commonly involved in the pathogenesis of mesothelioma. PMID- 12894505 TI - The antitumor ether lipid 1-Q-octadecyl-2-O-methyl-rac-glycerophosphocholine (ET 18-OCH3) inhibits the association between Ras and Raf-1. AB - BACKGROUND: Previous studies have shown that the antitumor ether lipid, 1-O Octadecyl-2-O-methyl-rac-glycerophosphocholine (ET-18-OCH3), inhibits the activation of the MAPK pathway in EGF- and serum-stimulated MCF-7 cells. The activation of the MAPK pathway subsequent to growth factor stimulation requires the recruitment of Raf-1 from the cytosol to the membrane. ET-18-OCH3 decreased the level of membrane-associated Raf-1 relative to untreated control cells. Since ET-18-OCH3 did not inhibit the activities of the kinases in the cascade, the reduced Raf-1 levels appeared to be the cause for the reduction in the magnitude and duration of MAPK activity. In this study we have investigated whether the reduced Raf-1 levels arise from a perturbation of the interaction of Raf-1 with activated Ras, which is the event that mediates the membrane recruitment. MATERIALS AND METHODS: The interaction of Raf-1 with Ras was examined by investigating the association of cytosolic Raf-1, from ET-18-OCH3-treated and untreated cells with purified GST-Ras-GTP-gamma-S bound to agarose beads. The level of associating Raf was determined by Western blot analysis. The effect of naturally occurring phospholipids on the Raf-1-Ras interaction was also examined to assess the specificity of the results. RESULTS: In cells preincubated with ET 18-OCH3, the interaction of GST-Ras-GTP-gamma-S with cytosolic Raf was reduced. The addition of ET-18-OCH3 to the cytosolic fraction isolated from untreated cells also reduced the binding of Raf to activated GST-Ras-GTP-gamma-S. Cytosolic Raf-1 from cells incubated with natural lysophospholipids was similar to that of controls. CONCLUSION: Our findings suggest that ET-18-OCH3 associates specifically with Raf-1 in the cytosol and interferes in the interaction of Raf-1 with activated Ras, thereby reducing the levels that are translocated to the membrane for activation. Thus Raf-1 appears to be a molecular target of ET-18 OCH3. PMID- 12894506 TI - The inhibitory effect of matrix metalloproteinase inhibitor ONO-4817 on lymph node metastasis in tongue carcinoma. AB - BACKGROUND: ONO-4817 is a novel matrix metalloproteinase (MMP) inhibitor which has a broad inhibitory spectrum for MMPs. We investigated the inhibitory effect of ONO-4817 on cervical lymph node metastasis of tongue carcinoma. MATERIALS AND METHODS: HSC-3-M3, a cell line derived from human tongue carcinoma with high metastatic potential for lymph node, was used. The direct cytotoxicity of ONO 4817 was assessed by MTT assay and the gelatinolytic activity by gelatin zymography and film in situ zymography (FIZ). The inhibitory effect on lymph node metastasis was examined by orthotopic implantation model with nude mice. RESULTS: ONO-4817 had no direct cyototoxicity on HSC-3-M3 cells. Gelatin zymography and FIZ demonstrated a suppression of MMP-9 activation and a marked inhibition of gelatinolysis. Additionally, a suppression of cervical lymph node metastasis was shown by therapeutic experiment. CONCLUSION: The MMP inhibitor, ONO-4817, is a candidate for adjunctive therapy of cervical lymph node metastasis in tongue carcinoma. PMID- 12894507 TI - The effect of cyclooxygenase inhibitor diclofenac on experimental murine colon carcinoma. AB - BACKGROUND: Cyclooxygenase-2 (Cox-2) has been found to be overexpressed in several types of human cancers and its role in tumorigenesis has been proposed. The aim of this study was to investigate the effects of the cyclooxygenase inhibitor diclofenac on the growth of murine C-26 colon carcinoma cells. MATERIALS AND METHODS: Expression of Cox-2 mRNA and protein was examined by RT PCR analysis and immunohistochemistry, respectively. By using MTT-assay, we examined the effects of diclofenac at various concentrations on the growth of C 26 cells in vitro. The effect of diclofenac on the growth of the C-26 tumor in syngeneic mice was also investigated. RESULTS: By RT-PCR, Cox-2 mRNA was detected in C-26 cells. Cox-2 protein was localized to C-26 cells and treatment with diclofenac resulted in apoptotic cell death in a dose-dependent manner. Diclofenac administered in drinking water resulted in growth inhibition of C-26 tumor in mice and correlated with plasma levels of both PGE2 and TXB2. CONCLUSION: Our data show that diclofenac may be a potential agent for the prevention and treatment of human colon cancer. PMID- 12894508 TI - Enhancement of Fas-ligand-mediated programmed cell death by taurolidine. AB - BACKGROUND: Taurolidine was recently found to have a direct and selective antineoplastic effect on brain tumor cells. The ability of taurolidine to exert antineoplastic action by enhancement of Fas-mediated apoptosis in different malignant glioma cell lines was investigated. MATERIALS AND METHODS: Human derived U373 cells were cultured and incubated with taurolidine and the median inhibitory concentration (IC50) was calculated. Flow cytometric analysis was performed to assess changes in DNA content. The cells were qualitatively and quantitatively examined using light microscopy and electron microscopy. LN-18 and LN-229 cells were incubated in the absence or presence of either Fas-ligand, taurolidine or respective combinations thereof. The cell viability was determined by adding a double concentrated WST-1 reagent. The activity of the mitochondrial succinate reductase was measured in an ELISA reader. RESULTS: The exposure of U373 cells to taurolidine led to a concentration-dependent (IC50 35.8 +/- 2.2 micrograms/ml) loss of cell viability. Flow cytometric analysis demonstrated a concentration-dependent appearance of DNA debris in the sub-Go/G1 region. In the presence of 6.25 vol.% Fas-ligand, LN-18 cells displayed more than 90% loss of cell viability, whereas the viability of LN-229 cells was reduced only at higher concentrations of Fas-ligand. Taurolidine by itself did not appreciably affect the viability of LN-18 cells in the investigated concentration range, but was able to enhance the effect of Fasligand on LN-18 cells. The exposure of LN-229 cells to taurolidine alone caused an appreciable loss of cell viability by about 70% at the highest concentration tested. Cell destruction by Fas-ligand (10 vol.%) was enhanced in the presence of taurolidine. CONCLUSION: The antineoplastic activity of taurolidine seems to be partially based on the enhancement of Fas-ligand-induced apoptosis. In addition, taurolidine was demonstrated to have an antieoplastic effect independent of Fas-ligand. Perhaps taurolidine exerts antineoplastic activity based on different mechanisms. PMID- 12894509 TI - EGFR but not PDGFR-beta expression correlates to the antiproliferative effect of growth factor withdrawal in glioblastoma multiforme cell lines. AB - BACKGROUND: The aim of the current study was to investigate a putative relationship between (i) growth characteristics (proliferation and tumorigenicity) of nine glioblastoma multiforme (GBM) cell lines under different growth-stimulating conditions in vitro and (ii) their basal expression of a panel of growth factor receptors/autocrine cytokines. MATERIALS AND METHODS: Basal expressions of the epidermal growth factor receptor (EGFR), platelet-derived growth factor receptor-beta (PDGFR-beta), platelet-derived growth factor-AA (PDGF AA) and PDGF-BB, tumor growth factor-alpha (TGF-alpha) and TGF-beta as well as tumor necrosis factor-alpha (TNF-alpha) were determined by immunocytochemistry at standard cell culture conditions (10% fetal calf serum [FCS]). Proliferation and tumorigenicity at 10% FCS and relative serum starvation (0.5% FCS) were assessed by using Coulter counting and soft agar cloning, respectively. RESULTS: The ratio between cell multiplications at 10% and 0.5% FCS over a 10-day period was defined as a measure of growth factor dependence of cellular proliferation. Expression of EGFR (but not of PDGFR-beta) strongly correlated to this ratio (Spearman rank correlation coefficient R = 0.87). No considerable correlations were present among other appropriate pairs of variables with biologically founded putative relationships. CONCLUSION: Greater expression of EGFR is associated with increased growth factor dependence of cellular proliferation. Our results strengthen the role of EGFR as a rational molecular target of therapeutic intervention in GBM. PMID- 12894510 TI - A common pathway for chemotherapy-induced apoptosis in human squamous cell carcinoma lines distinct from that of receptor-mediated cell death. AB - Regulation of apoptosis by extracellular molecules binding to cell death receptors has received much attention in recent years. Fas, a member of the tumor necrosis factor receptor superfamily, is a transmembrane protein whose extracellular domain binds its cognate ligand (FasL), which can induce apoptosis in sensitive cells. Fas ligation leads to activation of cell death proteases, thereby initiating a proteolytic cascade which results in cellular fragmentation and death. Apoptosis is also regulated by inhibitory signals which promote cell survival. The bcl2 family of proteins is composed of both inhibitors and activators of programmed cell death. The bcl2 protein itself inhibits many apoptotic stimuli while other members of the bcl2 family such as bak and bid promote cell death. Many types of cancer chemotherapy induce cellular stress leading to induction of apoptosis. Stress-activated protein kinases such as p38 have been shown to inactivate bcl2 through phosphorylation and induce cleavage of bid. Deficiency of proapoptotic bcl2 family members has been associated with drug resistant phenotypes. We report that exposure of human squamous cell carcinoma lines to different chemotherapy drugs activates a caspase cascade which is distinct from that of receptor-mediated apoptosis. The variable sensitivity of each cancer cell line to different forms of chemotherapy was not due to differences in caspase or bcl2 family protein expression. Rather, the stress activated protein kinase p38 was overexpressed by resistant SCC lines which correlated with reductions in proapoptotic bid and bak protein expression. These two proteins exhibit distinct patterns of intracellular localization during chemotherapy-induced apoptosis. PMID- 12894511 TI - Expression of stem cell factor and c-kit in human malignant fibrous histiocytoma cell line (TNMY1). AB - BACKGROUND: The expression of c-kit and/or its ligand, stem cell factor (SCF), has been related to tumor proliferation, in several tumor systems. MATERIALS AND METHODS: We analyzed the expression of the SCF/its receptor (c-kit) mRNA and the production of soluble SCF in a human malignant fibrous histiocytoma (MFH) cell line (TNMY1). RESULTS: Immunocytochemical analysis revealed that the TNMY1 cells were positive for both SCF and c-kit. Reverse transcriptase-polymerase chain reaction (RT-PCR) showed that the TNMY1 cell line expressed mRNA for SCF and c kit. By using an enzyme-linked immunosorbent assay (ELISA), the TNMY1 cells were found to produce relatively high amounts of soluble SCF. However, the addition of soluble SCF to the TNMY1 cells did not alter the in vitro growth ability of the cells. CONCLUSION: Our data showed that the MFH cells produced consistent amounts of SCF but did not demonstrate autocrine growth modulation. Thus, SCF secretion may have a paracrine activity in the growth of MFH cells. PMID- 12894512 TI - Decreased expression of cyclin G2 is significantly linked to the malignant transformation of papillary carcinoma of the thyroid. AB - BACKGROUND: Cyclin G2 is a novel cyclin negatively regulating the cell cycle progression, contrary to the characteristics of conventional cyclins. However, little is known about the cyclin G2 expression in human carcinomas. We thus investigated cyclin G2 expression in human thyroid neoplasms. MATERIALS AND METHODS: We immunohistochemically examined cyclin G2 expression in 40 normal thyroids and 80 thyroid neoplasms. RESULTS: Normal thyroids expressed cyclin G2 in more than 5% of follicular cells. Of 30 papillary carcinomas including 6 microcarcinoma, cyclin G2 expression was not, or only occasionally, observed in carcinoma cells, indicating its expression decreased in all these cases. On the other hand, in 16 of the 24 follicular adenomas (66.7%) and 5 of the 23 follicular carcinomas (21.7%), cyclin G2 expression was retained (more than 5% of neoplastic cells were positive), and adenomas more frequently (p = 0.0032) retained cyclin G2 expression than carcinomas. CONCLUSION: Our results suggest that lack of cyclin G2 plays an important role in the malignant transformation of papillary carcinoma. Also, it may play an adjuvant role in the transformation of follicular adenoma to carcinoma. This is the first study of the expression of cyclin G2, a novel cyclin having a role opposite to that of conventional cyclins, in human carcinoma. PMID- 12894513 TI - Expression profiling of tumor necrosis factor alpha-induced apoptosis-associated genes in human solid tumor cell lines. AB - BACKGROUND: The induction of genes associated with cellular apoptosis by tumor necrosis factor-alpha (TNF-alpha) in human cancer cell line sof various tissue origins may characterize TNF-alpha responder cell lines/cancers. MATERIALS AND METHODS: Using quantitative real-time polymerase chain reaction (PCR), the comprehensive molecular profiling of genes downstream of the TNF-alpha receptor genes in 91 well-defined human cancer cell lines allowed us to elucidate relationships between TNF-alpha response and the genetic expression profiles of the target cell lines. RESULTS: Among the 52 genes tested, the above average expression of Akt mRNA showed significant correlation with TNF-alpha-induced susceptibility to apoptosis. In addition, multidrug resistance protein 5 (MRP5) and tumor necrosis factor receptor type 1 (TNFR1) mRNA expressions also appear to be possible markers for responsiveness to TNF-alpha. CONCLUSION: These results provide a preliminary basis for the screening for genetic markers that may help to predict a favorable therapeutic outcome, and also to identify patients who may benefit from cytokine therapy. PMID- 12894514 TI - Her2/neu expression in poorly-differentiated and anaplastic thyroid carcinomas. AB - The Her2/neu expression in poorly-differentiated thyroid carcinomas (PDTC) and in anaplastic thyroid cancer (ATC) was investigated retrospectively. Tumors of 25 patients suffering from PDTC and 25 from ATC were evaluated, including 15 cases of PDTC with highly-differentiated tumor parts. Her2/neu levels were immunohistochemically detected on formalin-fixed, paraffin-embedded tissues using c-erbB-2/Her2/neu Oncoprotein Ab-17 monoclonal antibody(mAb). In statistical analysis using 1-way ANOVA, Her2/neu protein overexpression was highly significantly correlated with the differentiation status of the investigated tumor parts. Whereas ATCs showed only few positive tumor cells, PDTCs presented with large reacting tumor areas and highly-differentiated parts of PDTCs revealed the highest staining intensity. These findings suggest a decreasing impact of Her2/neu oncoproteins correlating to dedifferentiation, reflecting the aggressive biological behaviour of these tumors. PMID- 12894515 TI - A novel porphyrin photosensitizer from bamboo leaves that induces apoptosis in cancer cell lines. AB - BACKGROUND: In the screening of new anticancer agents, we found that a methanol extract of bamboo leaves induced rapid apoptosis in the human leukemia CMK-7 cell line. MATERIALS AND METHODS: The active compounds in the extract were isolated by chromatographic methods and their structures were determined by NMR and mass spectroscopy. Apoptosis by the compounds were evaluated in CMK-7 and human colon adenocarcinoma Colo320 DM cells by monitoring the caspase-3 activation and DNA cleavage. RESULTS: The active compounds are 201-hydroxypurpurin-7 delta-lactone ethyl methyl diester (1) and the corresponding methyl phytyl diester (2). The apoptosis by compound 1 (0.3 to 0.1 microM for CMK-7 cells) was enhanced when the culture was briefly irradiated with a fluorescent lamp. This photodynamic induction of apoptosis by compound 1 was much stronger than that by a known photosensitizer, pyropheophorbidemethyl. Compound 2 was a weaker inducer of apoptosis than compound 1, but the apoptosis occurred after light irradiation. CONCLUSION: The 201-hydroxypurpurin-7 delta-lactone esters are promising lead compounds as photosensitizers for photodynamic therapy of cancer. PMID- 12894516 TI - Modulation by the ATP/GTP ratio of the phosphorylation level of P-glycoprotein and of various plasma membrane proteins of KB-V1 multidrug resistant cells. AB - The level of protein phosphorylation is known to affect the properties of various membrane proteins. We have previously shown that GTP is capable of greatly enhancing the phosphorylation by [gamma-32P]ATP of P-glycoprotein (Pgp) from KB V1 cells (3). Investigating the possibility of a general modulation of [gamma 32P]ATP plasma membrane protein phosphorylation, we found that phosphorylation of other membrane proteins are also modulated by various combinations of [ATP + GTP]. The ATP/GTP ratio giving the highest phosphorylation level depended on the protein studied. Modulation of the [gamma-32P]ATP-mediated phosphorylation of numerous membrane proteins requires hydrolysis of both ATP and GTP. ADP and GDP also increased [gamma-32P]ATP-driven phosphorylation but to a lesser extent than GTP. This plasma membrane endogenous phosphorylation activity was neither inhibited by specific inhibitors of protein kinase C, nor by inhibitors of cAMP- or cGMP-dependent protein kinases or of casein kinase II, respectively. Mastoparan, a G-protein regulator, increased the phosphorylation of some proteins that were already enhanced by the presence of [ATP + GTP] mixtures, especially proteins migrating in gels at the same position as P-glycoprotein. PMID- 12894517 TI - Detection of paraneoplastic anti-neuronal-specific antibodies: comparison of different immunohistochemical techniques. AB - Different immunohistochemical techniques have been employed to identify the anti neuronal antibodies in patients with paraneoplastic neurological syndromes. The finding of anti-neuronal-specific autoantibodies in serum or cerebrospinal fluid well correlates with particular types of tumors, thus leading, in many cases, to an early cancer identification. In this work, we have compared three immunohistochemical methods (immunofluorescence, indirect immunoperoxidase and avidin-biotin immunoperoxidase) on frozen and paraffin-embedded sections of rat cerebellum in order to set up a reliable and simple technique for the diagnosis of these syndromes. Our study demonstrates that the best result was obtained by using frozen sections of rat cerebellum and the avidin-biotin immunoperoxidase method that also allowed the identification of anti-GAD antibodies not detected in paraffin-embedded tissues. PMID- 12894518 TI - Growth-inhibitory effect of phosphatidylinositol 3-kinase inhibitor on human pancreatic cancer cells and expression of Bcl-2 family. AB - Pancreatic cancer is one of the most devastating malignant tumors in humans and novel modalities for treatment need to be developed. We studied the mechanism of the growth-inhibitory effect of phosphatidylinositol 3-kinase (PI 3-K) inhibitor on human pancreatic cancer cells from the point of view of expression of the Bcl 2 family proteins. Growth of AsPC-1 and COLO-357 human pancreatic cancer cells was inhibited by a phosphatidylinositol 3-kinase (PI 3-K) inhibitor, wortmannin, and this growth-inhibitory effect was more marked in medium containing 10% fetal bovine serum (FBS) than in serum-free medium. In these cells, DNA fragmentation increased with the concentration of wortmannin in a dose-dependent manner. In Panc-1 human pancreatic cancer cells, cell growth and induction of DNA fragmentation were not influenced by treatment with wortmannin at concentrations up to 25 microM. Western blot analysis showed a decrease in expression of BclXL protein in AsPC-1 and COLO-357 cells by treatment with 25 microM wortmannin and this decrease was especially prominent in AsPC-1 cells. On the other hand, the expression of BclXL protein in Panc-1 cells was not influenced by treatment with wortmannin. The expression of BclXS protein was not detected by conventional Western blotting and the expression of Bcl-2 and Bax protein was not altered by wortmannin in all three cell lines. Decrease in expression of BclXL protein could be partly involved in the growth-inhibitory effect of the PI 3-K inhibitor, wortmannin, on pancreatic cancer cells. Although the growth of Panc-1 cells was not inhibited by wortmannin, PI 3-K inhibitor could still be one of the candidates for treatment of pancreatic cancer and BclXL could be a target for gene therapy. PMID- 12894519 TI - Serum HHV8 DNA and Tat antibodies in Kaposi's sarcoma patients with and without HIV-1 infection. AB - Human herpesvirus 8 or Kaposi's sarcoma-associated herpesvirus (HHV8/KSHV) is believed to be the most important etiopathological factor of Kaposi's sarcoma (KS) and some specific types of malignant lymphomas. The diagnostic and prognostic significance of serum viral load in endemic (African) areas is poorly understood. In AIDS-related KS (AKS) it has been shown that HIV-Tat may be of pathogenic importance and that immunoreactivity to Tat may have prognostic significance. Here we report on the quantitative analysis of HHV8 DNA in serum from Tanzanian patients with KS (n = 19), either AIDS-related (AKS) (n = 14) or endemic KS (EKS) (n = 5) and non-KS control individuals (n = 4). Fourteen AKS sera were also tested for HIV-tat antibodies by a direct ELISA assay. In AKS patients detectable (12 out of 14) serum HHV8 DNA levels showed a median of 1400 copies/ml as compared to a median of 200 copies/ml for EKS, but for one AKS case with an exceptionally high level (25,500 copies/ml). The serum HHV8 DNA levels were usually higher in males (n = 17; median 580 DNA copies/ml) as compared to females (n = 6; median 120 DNA copies/ml) and in early, patch stages (n = 8; median 2,750 copies/ml) as compared to late, nodular stages (n = 11; median 200 DNA copies/ml). Of fourteen sera from AKS patients, seven were positive for antibody against HIV-1 tat. Epitope analysis of the anti-tat antibody spectrum showed reactivity to various non-functional sites, but not towards the functional epitopes 46-60 (TAR-binding region). PMID- 12894520 TI - Sodium iodide symporter/radioactive iodine system has more efficient antitumor effect in three-dimensional spheroids. AB - The sodium iodide symporter (NIS) is a plasma membrane protein that mediates active uptake of inorganic iodide from plasma into thyroid cells. Expression of the NIS gene in tumor cells may provide a novel mechanism for treating cancer. Previously, we cloned cDNA of rat NIS into the retroviral vector LXSN, transduced human and murine tumor cells, and demonstrated 50-60% killing effect of 131I in cells expressing NIS. However, monolayer cultures of cancer cells cannot adequately represent some aspects of the growth and microenvironmental conditions of three-dimensional (3D) solid tumors. On the other hand, animal models are time consuming and costly and not always reasonable for the first estimation of scientific approaches. An in vitro model of multicellular tumor spheroids growing as a 3D-structure provides an important link between monolayer cell cultures and animal experiments. This model is especially crucial for suicide systems utilizing radioisotopes as the killing agent because the deposition of radiation energy causes DNA damage in cells more effectively in 3D-structure. In this study we used a retroviral vector for introduction of the rat NIS gene into DU145 prostate cancer cells. The liquid-overlay technique was exploited to create a tumor spheroid model. We established that uptake of 125I in monolayer culture cells containing the NIS was 40- to 50-fold greater than in DU145 cells without NIS. Using a clonogenic assay for monolayer culture, we demonstrated a 50-70% killing effect of 131I on DU145 cells expressing the NIS gene. The same dose of 131I resulted in complete death of tumor spheroids composed of the DU145-NIS cells. Our data demonstrates significant anti-tumor efficacy of novel radioisotope concentrator gene therapy in a multicellular spheroid model, thus proving to be a useful link between the in vitro system and in vivo animal model. PMID- 12894521 TI - Combination therapy with paclitaxel and thalidomide inhibits angiogenesis and growth of human colon cancer xenograft in mice. AB - BACKGROUND: Combination chemotherapy is increasingly practiced for treating malignancies with greater sensitivity and less toxicity. Paclitaxel is a potent anti-tumor agent but has dose-limiting side-effects, whereas thalidomide is an orally active anti-angiogenic drug but less than sufficient to exert anti-tumor effect as a single agent. MATERIALS AND METHODS: Nude mice bearing hypervascular (LS174T) and less vascular (HT29) colon carcinomas were challenged with either a non-cytotoxic dose of paclitaxel, thalidomide or a combination of paclitaxel and thalidomide. RESULTS: Significant growth retardation was noticed only in the combination treatment group of LS174T tumors. Microvessel density data indicated a significantly low count in the combination treatment group compared to the others. Trends of decreased expression of angiogenic growth factors and increased apoptotic index were noticed in the combination treatment group. CONCLUSION: The results of this study underscore the therapeutic efficacy of concomitant use of paclitaxel and thalidomide in the treatment of highly vascular colorectal tumors in a xenograft model. PMID- 12894522 TI - Induction of early apoptosis marker by morphine in human lung and breast carcinoma cell lines. AB - We have investigated whether morphine and codeine, potent analgesic compounds most commonly used as cancer pain relievers, show tumor-specific cytotoxic activity and whether they can induce apoptosis or necrosis by monitoring the stainability with Annexin V and propidium iodide with fluorescence-activated cell sorter. Both opioids showed higher cytotoxic activity against three human tumor cell lines (lung carcinoma A549, mammary gland carcinoma MCF7, promyelocytic leukemia HL-60) than against three normal human cells (periodontal ligament fibroblast HPLF, gingival fibroblast HGF, pulp cell HPC). Morphine produced the major part of the apoptotic cell populations and the minor part of the necrotic cell populations in A549 and MCF7 cells, more effectively than codeine. In addition, morphine increased the activity of mitochondrial Mn-containing superoxide dismutase (MnSOD) in HL-60 cells, but decreased the MnSOD activity in A549 and MCF7 cells. The apoptosis-inducing activity of opioids may provide new strategies for the treatment and prevention of cancer. PMID- 12894523 TI - Growth response of colon cancer cell lines to selective estrogen receptor modulators. AB - BACKGROUND: The purpose of this study was to compare the "in vitro" effects of the selective estrogen receptor modulators, tamoxifen and raloxifene, on two human colon cancer cell lines. MATERIALS AND METHODS: Serial concentrations (0.1, 1, 5 and 10 microM) of tamoxifen and raloxifene were used and evaluated for cell proliferation, viability and apoptosis in HCT8 and HCT116 cells. RESULTS: Micromolar doses of raloxifene significantly reduced HCT116 and HCT8 cell proliferation. Tamoxifen (5 microM) strongly reduced HCT8 cell growth with minor effects on HCT116 cells. Raloxifene (10 microM) was lethal on both cell lines, while 10 microM tamoxifen caused lethality only in HCT8 cells. Five microM raloxifene reduced cell viability in HCT8 and HCT116 cells, while 5 microM tamoxifen halved only HCT8 cell viability. Raloxifene and tamoxifen did not induce apoptosis in the two cell lines. CONCLUSION: Tamoxifen, and even more raloxifene, were effective in reducing HCT8 and HCT116 cell proliferation and viability, suggesting their potential application in the prevention and therapy of colorectal cancer. PMID- 12894524 TI - The thioredoxin-thioredoxin reductase system: over-expression in human cancer. AB - Redox control has emerged as a fundamental biological control mechanism. One of the major redox control systems is the thioredoxin system comprised of thioredoxin (TRX) and thioredoxin reductase (TR). Together they form a powerful system involved in many central intracellular and extracellular processes including cell proliferation, the redox regulation of gene expression and signal transduction, protection against oxidative stress, anti-apoptotic functions, growth factor and co-cytokine effects, and regulation of the redox state of the extracellular environment. Over recent years this system has increasingly been linked to the development and expression of cancer phenotypes. In this report immunocytochemical approaches have been used to simultaneously determine the expression and localisation of both TRX and TR in primary human cancers, including breast cancer, thyroid, prostate and colorectal carcinoma, and malignant melanoma. In aggressive invasive mammary carcinomas and advanced malignant melanomas, thioredoxin was highly over-expressed compared to tumours of lesser aggressive nature. TRX expression was found in both nuclear and cytoplasmic location in the neoplastic cells. Furthermore, increased levels of TRX positively correlate with thioredoxin reductase (TR) expression and localisation. These results, which are the first immunocytochemical studies on the in vivo expression and localisation of TRX and TR in melanomas, thyroid, prostate and colorectal carcinomas and the first reports of TR expression in breast carcinomas, significantly extend the range of human cancers for which such data is available. Overall the results support the conclusion that aggressive tumours greatly over-express both TRX and TR. Such tumours have a high proliferation capacity, a low apoptosis rate and an elevated metastatic potential strongly implicating the involvement of the TRX system in the processes of oncogenesis and tumourogenesis and confirming its potential as a target for anticancer therapy for a wide range of human tumours. PMID- 12894525 TI - Growth factors regulate the expression profile of their syndecan co-receptors and the differentiation of mesothelioma cells. AB - BACKGROUND: Diffuse malignant pleural mesotheliomas are locally aggressive and highly lethal tumors that are becoming more common. The tumor derives from pluripotential mesothelial stem cells, which differentiate into epithelial or mesenchymal elements. Tumors with a predominantly epithelial growth pattern have a better prognosis than the sarcomatous and mixed types, the phenotype being important for the biology of the tumor. We have previously shown that mesotheliomas express a wide range of cell surface heparan sulfate proteoglycans (HSPGs), particularly syndecans, which interact with growth factors and matrix components. MATERIALS AND METHODS: This study was undertaken to examine the epithelial-mesenchymal transition of mesothelioma cells by exposing epithelially differentiated cells to an array of growth factors. Following substitution with TGF-beta 2, EGF, FGF-2, IGF-I and PDGF-BB, the expression levels of syndecans-1, 2 and -4 were monitored at selected times (30 minutes, 6 hours and 18 hours) by semi-quantitative RT-PCR and FACS analysis. The morphological appearance and proliferation rate of the treated cells was correlated to the PG profile obtained and to the subcellular compartmentalization of PGs. RESULTS: An early response was obtained only for syndecan-4. Changes in the differentiation pattern appeared later. Exposure to EGF and IGF-I induced a fibroblast-like morphology, simultaneously with a reduced expression of syndecans-1 and 2. TGF-beta 2 enhanced the focal contacts and showed a marked up-regulation of syndecan-4 and down-regulation of syndecan-1. Interestingly, TGF-beta 2 delayed the nuclear transport of syndecan-1 concomitantly with an antiproliferative effect. CONCLUSION: Growth factor signalling seems to be delicately controlled by an autoregulatory loop involving the syndecan expression levels and amounts of soluble HS chains shed into the medium. PMID- 12894526 TI - Doxorubicin-derived metabolites induce release of cytochrome C and inhibition of respiration on cardiac isolated mitochondria. AB - BACKGROUND: The generation of doxorubicin metabolites other than semiquinone free radicals and mitochondrial dysfunction have been implicated in doxorubicin (DOX) induced cardiotoxicity. This study examines pro-apoptotic mechanisms in isolated rat cardiac mitochondria exposed to DOX-derived cardiac metabolites i.e. doxorubicinol, doxorubicin aglycone and doxorubicinol aglycone. MATERIALS AND METHODS: Freshly isolated mitochondria were incubated in the presence of doxorubicin and its derivatives and the released cytochrome c was detected by Western blotting analysis. Oxygen consumption was measured with a Clark-type oxygen electrode and mitochondrial transmembrane potential (delta psi) was measured in a fluorometer in the presence of a fluorescent probe. RESULTS: The data obtained show that exposure of isolated mitochondria to the drugs determines a significant release of cytochrome c and a slight decrease in the mitochondrial transmembrane potential (delta psi). These effects are more evident when the experiments are performed in the presence of the aglycone derivatives. Moreover, an inhibition of the respiratory chain at the level of complex I is evidenced in the drug-treated mitochondria. CONCLUSION: The data obtained are consistent with the proposal that doxorubicin-induced cardiotoxicity may be partially exerted by the induction of programmed cell death, both directly and even more through its derived metabolites. PMID- 12894527 TI - In vivo gadolinium neutron capture therapy using a potentially effective compound (Gd-BOPTA). AB - BACKGROUND: In a previous study, we found that Gadobenate dimeglumine (Gd-BOPTA) resulted in a significantly greater Gd uptake by brain tumor tissue than Gadopentate dimeglumine (Gd-DTPA). Therefore, we investigated whether Gd-BOPTA is an efficient agent for neutron capture therapy (NCT). MATERIALS AND METHODS: Four groups of Fisher344 rats (control, neutron (n), n+ Gd-DTPA, n+ Gd-BOPTA) were subcutaneously injected 9L gliosarcoma cells in both hind legs. Gd-BOPTA and Gd DTPA (0.05 mmol/g tumor weight) were injected directly into the tumor. At the peak of Gd uptake, thermal neutron irradiation was applied. RESULTS: Two Gd+ groups showed pronounced tumor growth delay as compared with the control and neutron groups (p = 0.0053, 0.0064, respectively). Furthermore, the BOPTA group showed significantly prolonged delay of tumor growth as compared to the DTPA group (p = 0.033). CONCLUSION: This is the first report of Gd-NCT to demonstrate that Gd-BOPTA serve as an effective compound for NCT. Better cytocidal effects of Gd-BOPTA warrant further investigation of subcellular Gd distribution. PMID- 12894528 TI - Dendritic cell maturation induced by delivery of ultraviolet-mediated apoptotic colorectal cancer cell lines. AB - BACKGROUND: Dendritic cells (DCs) play a critical role in the immune response. The aim of this study was to investigate apoptotic tumor cells as an antigen source for DC maturation. MATERIALS AND METHODS: We compared the efficacy of ultraviolet (UV)-irradiation with that of gamma-irradiation in the induction of apoptosis of tumor cells. Phenotypic and functional changes of DCs were analyzed after co-incubation with UV-irradiated tumor cells. RESULTS: UV-irradiation (1.8 J/cm2) was a more reliable method of inducing apoptosis than gamma-irradiation (20,000 rad). The expression of costimulatory molecules on DCs was upregulated after co-incubation with UV-irradiated tumor cells. When we performed allogeneic mixed lymphocyte reaction assay, UV-irradiated tumor cells-pulsed DCs stimulated allogeneic T lymphocytes more efficiently than DCs pulsed with gamma-irradiated cells (HT29, p = 0.0419 and WiDr, p = 0.0076). CONCLUSION: UV-irradiation reliably induces apoptosis in cultured colorectal cancer cells. DCs mature in function and expression of costimulatory molecules after co-incubation with apoptotic tumor cells. The clinical implications warrant further study. PMID- 12894529 TI - In vitro and in vivo expression of somatostatin receptors in intermediate and malignant soft tissue tumors. AB - BACKGROUND: Previous studies showed that the subtype 2 of somatostatin receptors (sstr2) is generally expressed by soft tissue sarcomas. MATERIALS AND METHODS: Using reverse transcription-polymerase chain reaction (RT-PCR), the present study evaluated the expression of sstr mRNA in 31 human intermediate and malignant soft tissue tumors. In 8 patients somatostatin receptor scintigraphy was performed to detect soft tissue tumor sites and visualize in vivo sstr expression. RESULTS: Somatostatin receptors were identified in 84% of the tumors analyzed by RT-PCR analysis. sstr1 and sstr2 were the receptors most frequently detected. Somatostatin receptor scintigraphy was able to identify primary and/or metastatic sites in 7 out of 8 patients, with an overall sensitivity per site of 87%. Concordant results were found between in vitro and in vivo techniques. CONCLUSION: The wide expression of sstrs in soft tissue tumors suggest a relevant role for these receptors in diagnosis and may have therapeutic applications. PMID- 12894530 TI - Effects of treatment of rats with indole-3-carbinol on apoptosis in the mammary gland and mammary adenocarcinomas. AB - Induction of apoptosis is an approach to suppress carcinogenesis. The effects of a 12-week treatment of female Sprague-Dawley rats with indole-3-carbinol (I3C), beta-naphthoflavone or vehicle (40% ethanol in corn oil), by oral gavages starting 3 weeks after initiation of mammary tumorigenesis with 7,12 dimethylbenz[alpha]anthracene, on apoptotic activities in the mammary adenocarcinomas were examined. Apoptotic cells in tumor sections were detected by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling and quantitated by light microscopy and an Image-Plus Program. Activities of caspase 3, caspase-8 and caspase-9 were determined by colorimetric assays using the specific substrate and total tumor protein. There were no significant treatment related effects on the numbers of apoptotic cells and caspase activities in the mammary adenocarcinomas. Likewise, protein expression levels of Bcl-2 and Bax genes in these tumors, determined by Western blot analysis, showed no treatment related stimulation of apoptotic process. In the absence of tumorigenesis, the activities of caspase-3, caspase-8 and caspase-9 were increased up to approximately 3.6-fold in the mammary gland of rats treated with I3C at 5 or 25 mg/kg of body weight for 4 or 10 days. The I3C-effected induction of caspase-3 activity in the mammary gland was further confirmed by the cleavage of poly (ADP ribose) polymerase. Treatment of rats with 3,3'-diindolylmethane, a major product of I3C in vivo, at the dose levels equimolar to those of I3C above, did not increase the caspase activities in the mammary gland. Thus, this I3C dimer does not seem to account for the increases of apoptotic activities in the mammary gland observed with I3C. The results suggest that increase of apoptosis in the mammary gland induced by I3C before initiation of tumorigenesis may contribute to suppression of tumor development. PMID- 12894531 TI - Thalidomide down-regulates the expression of VEGF and bFGF in cisplatin-resistant human lung carcinoma cells. AB - Anti-angiogenic therapy represents one of the most promising treatment modalities for human cancer. Thalidomide (alpha-N-phthalimidoglutarimide) is a potent inhibitor of angiogenesis, and it is reported to overcome classical drug resistance in human multiple myeloma cells. However, the effect of this agent on the expression of angiogenic growth factors in cisplatin-resistant tumors is largely unknown. In the current study, we showed that thalidomide suppressed the expression of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) in cisplatin-resistant human A549DDP lung carcinoma cells. The mRNA levels of VEGF and bFGF were markedly decreased in the A549DDP cells treated with the therapeutic concentrations of thalidomide (0.6-6 micrograms/ml), as determined by RT-PCR analysis. Consistent with these results, thalidomide also significantly reduced the protein levels of VEGF and bFGF in these cells in a dose- and time-dependent manner. This study provided evidence to support the potential therapeutic applications of thalidomide in cisplatin-resistant human lung cancer and other tumors. PMID- 12894532 TI - Inhibition of proliferation and expression of AR/PSA by herbal supplement Equiguard in LNCaP cells cultured in androgen-proficient FBS and androgen deficient charcoal-stripped FBS is correlated with increased serine-15 phosphorylation of the tumor suppressor gene p53. AB - Use of dietary supplements and botanical products is widely accepted by patients diagnosed with prostate cancer (CaP) as a primary or complementary form of treatment for their medical conditions in the U.S. Yet, the majority of these products have not been rigorously studied with regard to scientific mechanism(s). Because many of the available products are mixtures of multiple extracts derived from plants, some of which are not necessarily native to the U.S., we consider mechanistic studies under defined laboratory conditions to be valuable and essential, not only from the standpoint of standardization and possible contamination with the products, but also in providing insights and scientific evidence for the clinical efficacy some of these products purportedly demonstrate. In previous studies from this laboratory, Equiguard, a composite supplement consisting of standardized extracts from nine Chinese herbs, which was originally formulated to correct physiological decline in kidney functions associated with age, was fortuitously found to display anti-CaP properties. Using a panel of CaP cells, we showed that ethanol extracts of Equiguard significantly inhibited cancer cell growth, induced apoptosis, lowered expression of the androgen receptor (AR), decreased intracellular and secreted prostate-specific antigen (PSA) levels and completely abolished the colony forming activities of CaP cells. Since responsiveness to Equiguard was observed in cells mimicking the androgen-dependent (AD) and androgen-independent (AI) states of CaP, our results raise the interesting possibility that this herbal supplement may potentially prevent, delay or circumvent the onset of AI, and thereby induce chronic instead of terminal CaP. Since androgen ablation therapy (chemical or surgical castration) is the mainstay for localized CaP, we questioned whether Equiguard might still exert the aforementioned activities in experimental settings modeled after androgen ablation. Accordingly, we studied the effects of Equiguard in LNCaP cells, cultured in androgen-proficient (FBS) or -deficient (CS-FBS) media that simulate the hormonal status pre- and post-castration in vivo. Extracts of Equiguard were effective in reducing colony formation, proliferation and PCNA expression of cells cultured in CS-FBS. Moreover, within a concentration range of Equiguard, the prostate-specific genes, PSA and AR, were affected to a similar extent in cells cultured either in FBS or CS-FBS, and were correlated with increased phosphorylation at serine-15 of the tumor suppressor gene p53. These results are consistent with the interpretation that the anti-proliferative and gene modulatory properties of Equiguard are largely independent of the status of androgens in the culture media. PMID- 12894533 TI - Irinotecan enhances IL-12 production by OK-432-activated murine macrophages. AB - BACKGROUND: Our previous study showed that the combination of irinotecan (CPT-11) and OK-432 had an additive antitumor effect. The purpose of this study was to analyze the mechanism by which this combined treatment had an effect on immunity. MATERIALS AND METHODS: To investigate the immune effects of murine splenocytes stimulated by SN-38 (the active form of CPT-11) and OK-432, endogenous interleukin (IL)-12 p70 production was assayed by ELISA and flow cytometry. RESULTS: Endogenous IL-12 production was increased by SN-38 stimulation of cultures of OK-432-activated splenocytes from C57BL/6, C3H and Balb/c mice, which was not observed with LPS-activated splenocytes. IL-12 production by splenocytes was higher at an early stage after tumor inoculation. SN-38 and OK-432 stimulated IL-12 production in cultures of peritoneal exudate macrophages (PEM), and T cell cooperation was essential in cultured splenocytes. CONCLUSION: These results suggest that the interaction of SN-38 and OK-432 may support a type 1 T helper (Th1)-dominant state through increasing endogenous IL-12 production, mainly by macrophages. PMID- 12894534 TI - Induction of apoptosis or necrosis by ionizing radiation is dose-dependent in MG 63 osteosarcoma multicellular spheroids. AB - This study investigates cell growth and death (apoptosis and necrosis) in actively proliferating MG-63 osteosarcoma spheroids in response to two doses of ionizing radiation (5 and 30 Gy). Cell growth was examined by growth curves and by cell cycle analyses utilizing flow cytometry. Death was examined in both disaggregated and whole spheroids by the chromatin dye Hoechst and by Western blot analysis of the bcl-2 family of proteins known to be involved in the apoptotic process. The results indicate that after exposure to 5 Gy MG-63 spheroids die by apoptosis, while after exposure to 30 Gy they die by necrosis. The analysis of the bcl-2 family of proteins demonstrates that bcl-2, bax and bcl XL are involved in triggering apoptosis in spheroids exposed to 5 Gy. The possibility of studying the mechanisms responsible for radiation-induced cell death in three-dimensional spheroids (which represent much more closely in vivo tumors) is extremely important if the foundations for more selective and controllable clinical radiotherapeutic protocols are to be laid. PMID- 12894535 TI - Novobiocin sensitizes BCRP/MXR/ABCP overexpressing topotecan-resistant human breast carcinoma cells to topotecan and mitoxantrone. AB - BACKGROUND: Novobiocin was shown to sensitize cancer cells to etoposide and alkylating agents. Human breast carcinoma cells exposed to topotecan (MCF7/TPT300 cells) developed resistance to both mitoxantrone and topotecan. An ATP-binding cassette family protein BCRP/MXR/ABCP was overexpressed in MCF7/TPT300 cells. In addition, topotecan efflux was markedly enhanced in the resistant cells. To investigate the possibility that novobiocin may enhance cytotoxicity in BCRP/MXR/ABCP overexpressing cells, we exposed MCF7/TPT300 cells to novobiocin. MATERIALS AND METHODS: Cytotoxicity tests of topotecan and mitoxantrone, as well as topotecan accumulation tests, were performed with or without novobiocin in MCF7/TPT300 cells. RESULTS: Novobiocin enhances topotecan and mitoxantrone toxicity in MCF7/TPT300 cells at a clinically relevant concentration. Novobiocin enhanced cellular accumulation of topotecan and inhibited topotecan efflux in MCF7/TPT300 cells. CONCLUSION: Novobiocin may enhance topotecan and mitoxantrone toxicity in topotecan-resistant breast carcinoma cells. Novobiocin may be useful to reverse topotecan or mitoxantrone resistance in the clinic. PMID- 12894536 TI - Analysis of MICA gene transcripts in human rectal cancers. AB - The human MHC class I chain-related gene A (MICA) encodes a protein which is an activator ligand for the NKG2D receptor on NK cells, CD8+ alpha beta T cells and gamma delta T cells. MICA expression is up-regulated upon cellular stress and its expression is correlated to infiltration of human NKG2D-bearing T cells into the tumors. It is assumed that the interaction of MICA-NKG2D ligand-receptor could play a significant role in induction of innate and adaptive responses against epithelial tumors, specifically those from the gastrointestinal tract. In this study MICA messenger RNA levels in human rectal carcinoma (Duke's stage B-D) and its normal adjacent tissue was analyzed in samples donated by 18 patients undergoing rectal tumor resection. Quantitative RT-PCR analysis from rectal tumors revealed that the overall expression of MICA at mRNA level differs extensively among individual tumors. In addition, invasive rectal tumors tend to up-regulate MICA whereas MICA mRNA levels were lower in early tumors. Differential transcription levels of MICA gene expression in rectal carcinomas at different stages is probably a strategy by tumors to escape confrontation with intraepithelial tumor-infiltrating T cells. PMID- 12894537 TI - Stress-related hormonal suppression of natural killer activity does not show menstrual cycle variations: implications for timing of surgery for breast cancer. AB - BACKGROUND: Evidence exists that prognosis in breast cancer may be related to the timing of primary surgery. Fluctuations in natural killer (NK) cells and/or their sensitivity to surgical stress hormones could mediate this phenomenon. We sought to establish, firstly, if there are any numerical or functional changes in NK cells during the course of the menstrual cycle and, secondly, if their prior in vitro incubation with cortisol and adrenaline produced functional change that was more pronounced at a particular point in the cycle. SUBJECTS AND METHODS: Normally menstruating females (n = 10) were sampled at two points (mid-follicular and mid-luteal) during two cycles. NK cell numbers were determined by flow cytometry. Natural killer cell activity (NKA) (unstimulated and following a 2 hour pre-incubation with cortisol and adrenaline) was determined using chromium 51 release assays from peripheral blood mononuclear cells (PBMCs) prepared from cryopreserved samples from the second cycle. RESULTS: NK cell numbers (expressed as a percentage of a standard lymphgate) were significantly higher at the mid luteal point compared to the mid-follicular point (8.45 +/- 3.71 vs. 6.85 +/- 2.80, p < 0.05, data pooled from both cycles). Corresponding changes in NKA were observed, although these did not reach statistical significance. The mean inhibitory effects of cortisol (67%) and adrenaline (12%) on NKA were significant, but this phenomenon was uninfluenced by cycle point. CONCLUSION: This study supports the role of menstrual cycle-induced alterations in NK cells as a putative mediator of improved survival when surgery for breast cancer is carried out in the follicular phase. However there is no evidence of the cyclical sensitivity of NK cells to the principal surgical stress hormones. PMID- 12894538 TI - Induction of apoptosis by combined treatment with differentiation-inducing agents and interferon-alpha in human lung cancer cells. AB - BACKGROUND: Differentiation-inducing agents for myeloid leukemic cells, such as dimethyl sulfoxide (DMSO), Na-butyrate and hexamethylene-bis-acetamide (HMBA), barely induce irreversible differentiation or growth inhibition in solid tumors when used alone. However, we previously reported that combined treatment with differentiation-inducing agents and interferon (IFN)-alpha effectively suppressed the growth of human lung cancer cell lines in vitro and in vivo. We show here that combined treatment-induced cell death is caused by apoptosis. MATERIALS AND METHODS: Induction of apoptosis was examined by expression of several apoptotic markers. RESULTS: Combined treatment-induced cell death is caused by apoptosis, which is characterized by the induction of apoptotic morphological changes and the activation of caspase-3-like protease. The expression of Apo2.7, an early apoptotic change, is also induced by this combined treatment, and is suppressed by the addition of Z-Asp-CH2-DCB, a pan caspase inhibitor. The signal transduction pathway of IFN-alpha is rapidly observed in lung cancer cells, although it does not induce significant growth inhibition when used alone. We analyzed the effect of IFN-alpha on the apoptotic pathway and found that IFN alpha but not DMSO induces the expression of caspase-4. The combination of IFN alpha and DMSO did not cause further increase in the expression of caspase-4. In many cases of the induction of apoptosis, cytochrome c is released from mitochondria, which induces the formation of large caspase-activating complexes. Caspase-3-like-activities induced by the combination of DMSO or Na-butyrate with IFN-alpha were analyzed by gel filtration and detected equally in fractions with molecular weights of 200-300 kDa, suggesting that caspase-3 is activated in large complexes in lung cancer cells. However, Na-butyrate and HMBA, when used alone, induced the release of cytochrome c and enhanced the loss of mitochondrial membrane potential, whereas DMSO had no effect. CONCLUSION: These results suggest that the combination of differentiation-inducing agents with IFN-alpha effectively induces apoptosis in lung cancer cells whereas the mechanism of such induction varies depending on the differentiation-inducing agent used. PMID- 12894539 TI - Application of semiempirical method to estimate the cytotoxic activity of gallic acid and its related compounds. AB - The semiempirical method was applied to estimate the cytotoxic activity of 9 gallic acid derivatives against human promyelocytic leukemic HL-60 cells. Positive correlation was found between their 50% cytotoxic concentration (CC50) value and the heat of formation or total energy, but not between the CC50 value and size of the molecule (determined either by the length of the side chains, the largest distance of the molecule, the distance from R2 to carbonyl C or benzene ring C, or by Van der waals area). When CC50 value was plotted vs octanol/water partition coefficient (log P), parabolic curve, with a bottom at log P = 3-4, was produced, in a similar fashion to vitamin K, prenylflavanone and eugenol-related compounds. The present study suggests the usefulness of the semiempirical method to estimate the cytotoxicity of structurally related compounds. PMID- 12894540 TI - Effects of glucocorticoids on antitumor effects of immunizations with fusions of dendritic and tumor cells. AB - BACKGROUND: A pitfall of immunotherapy with dendritic cells (DCs) for patients with malignant tumor is that certain patients have already been treated with Dexamethasone (DEX) prior to vaccination. Therefore, it is important to investigate whether DEX has negative effects on induction of antitumor immune response in dendritic cell-therapy. MATERIALS AND METHODS: We analyzed surface phenotypes and phagocytosis of DCs incubated with or without DEX. We also investigated the effects of DEX on in vivo antitumor effects of immunization with fusions of dendritic and glioma cells. RESULTS: Treatment with DEX did not affect the expression of surface markers, including H-2Kk, I-Kk, CD80 and CD86, on DCs. Likewise, DEX did not affect phagocytosis. Immunization with FCs inhibited tumor growth in vivo, and DEX did not suppress antitumor effects induced by FCs. CONCLUSION: These data suggest that vaccination with FCs can be used to treat cancer patients receiving DEX. PMID- 12894541 TI - Clinical efficacy of doxifluridine and correlation to in vitro sensitivity of anticancer drugs in patients with colorectal cancer. AB - We examined thymidine phosphorylase (TP) expression and sensitivity to anticancer drugs and compared the findings with the efficacy of 5'-deoxy-5-fluorouridine (5' DFUR), an intermediate metabolite of capecitabine. PATIENTS AND METHODS: Patients were enrolled in this study from January 1995 to June 1998 for stages II-III colorectal cancer with curative resection. We conducted sensitivity tests of tumor tissue to 5'-DFUR and 5-fluorouracil (5-FU) using the MTT method, and measured tumor tissue TP levels using enzyme-linked immunosorbent assay (ELISA). From 2 weeks postoperatively, the patients were given oral 5'-DFUR 800 mg/m2/day (5 days administration followed by 2 days discontinuation) for one year and they were followed for 3 years postoperatively. RESULTS: Of 139 patients registered, 124 were analyzed for the present study. The median 5'-DFUR administration was 362 days and the median total dose was 245.0 g. We compared prognoses in patients with positive and negative 5-FU sensitivity test results. There was a significantly better prognosis in 5-FU sensitivity-positive patients with stage III than that in the sensitivity-negative patients (p = 0.041). We also compared prognoses in patients with positive and negative 5'-DFUR sensitivity test results. There was no significant difference in cases with a cut-off value of 50% (p = 0.055), although patients with 5'-DFUR-positive-sensitivity tended to show longer survival. Patients with stage II and higher TP levels tended to have longer survival than those with lower TP expression, but there was no significant difference between groups (p = 0.087). The prognosis of patients with 5-FU positive sensitivity and higher TP levels, the positive group, tended to have longer survival than in the negative group, but there was no significant difference between groups (p = 0.083). CONCLUSION: 5'-DFUR sensitivity test results and TP values may predict the clinical effects of this drug in colorectal cancer. PMID- 12894542 TI - Restoration of gap junctional intercellular communication by dibutyryl-cAMP in renal epithelial cells treated with renal carcinogen. AB - Dimethylnitrosamine(DMN) is an alkylating agent and a known renal carcinogen. A short exposure of renal epithelial cells to cytotoxic concentrations of DMN influences the expression of gap junction proteins. In this study, we examined gap junctional intercellular communication and connexin 43 expression in renal epithelial cells treated with 1% DMN and also examined the effects of dibutyryl cAMP on preventing gap junctional disturbances. Connexin 43 becomes hypophosphorylated after treatment with 1% DMN for 15 minutes, but this hypophosphorylation is inhibited by pretreatment with dibutyryl-cAMP. These results suggest that changes in gap junction protein expression are early events associated with 1% DMN treatment of renal epithelial cells, and such changes are prevented by dibutyryl-cAMP pretreatment. PMID- 12894543 TI - Analysis of apoptosis signaling pathway in human cancer cells by codeinone, a synthetic derivative of codeine. AB - We have recently found that codeinone, an oxidation metabolite of codeine, induced apoptosis, characterized by internucleosomal DNA fragmentation and mitochondrial cytochrome c release in HL-60 human promyelocytic leukemic cell lines, most effectively among 10 opioids. These findings prompted us to investigate whether codeinone induces apoptosis in other human cancer cells and possible changes in mitochondrial enzyme. FACS analysis demonstrated that codeinone induced the production of ANNEXIN-positive apoptotic cells in three different human cancer cells (HL-60, MCF7, A549). The apoptotic cells were visualized by microscopical observation after staining with Hoechst (H)-33342. Fluorometric assay showed that codeinone time-dependently activated caspase 3 and caspase 9, but not caspase 8, suggesting the activation of intrinsic apoptotic signaling pathway via mitochondria. Western blot analysis demonstrated that codeinone enhanced the Pro-apoptotic Bax protein expression, but reduced the anti apoptotic Bcl-2 protein expression. Codeinone did not significantly change the manganese superoxide dismutase (MnSOD) activity nor its mRNA expression. This apoptosis-inducing activity, in conjunction with antinociceptive activity, further substantiated the antitumor potential of codeinone. PMID- 12894544 TI - Carcinomas unresponsive to either cisplatinum or anti-EGFR therapy can be growth inhibited by combination therapy of both agents. AB - BACKGROUND: Previously we demonstrated that the antitumor efficacy of monoclonal antibodies against the EGFR (epidermal growth factor receptor) of human tumor xenografts mainly depends on the EGFR content of tumors rather than on the tumors' entity. In this study we wanted to elucidate whether the described cumulative effect of cisplatin and Anti-EGFR therapy also depends on the EGFR expression. MATERIALS AND METHODS: Xenotransplanted carcinomas with different EGFR levels were treated with monoclonal antibodies against the EGFR (EMD 72000 and EMD 55900), cisplatinum and a combination of both. RESULTS: Each monoclonal antibody alone led to an EGFR-dependent significant tumor growth reduction. Cisplatinum alone had no growth inhibitory effects on tumors with high content in contrast to those with low EGFR content. The combination of antibodies with cisplatinum resulted in an EGFR-independent tumor growth inhibition which was stronger than observed in the case of monotherapy. DISCUSSION: The obtained results may address upcoming phase I/II trials to use Anti-EGFR/Cisplatinum therapy regardless of the EGFR content of tumors. PMID- 12894545 TI - Overexpression of a novel lysyl oxidase-like gene in human head and neck squamous cell carcinomas. AB - BACKGROUND: To compare the mRNA expression profile of head and neck squamous cell carcinoma (HNSCC) cells and normal epithelial cells. MATERIALS AND METHODS: Differential display was used to trace genes showing differential expression in HNSCC cells. Rapid amplification of cDNA ends (RACE) was carried out to identify 5'upstream sequences followed by Northern hybridisation for verification of the differential mRNA expression. RESULTS: An overexpression of a 59 bp gene fragment was detected in HNSCC cells in contrast to normal epithelial cells. After cloning and sequencing the gene fragment, high grade of homology was found with a human cDNA full insert sequence (fis). After amplification and sequencing of the 5'end of the fis clone, subsequent database searches showed an exact match with the coding sequence of the human lysyl oxidase-like gene 4 (LOXL4). The differential expression of this 4.8 kb LOXL4-mRNA was confirmed by Northern hybridisation. CONCLUSION: The data presented in this work might emphasize the involvement of LOXL4 in molecular processes of the genesis or progression of head and neck carcinomas. PMID- 12894546 TI - Assessment of in vitro drug resistance of human breast cancer cells subcultured from biopsy specimens. AB - The low cellular yield of a breast cancer sample represents a limiting factor for in vitro chemosensitivity/chemoresistance testing. The use of in vitro serially cultured cells can help overcome this obstacle. MATERIALS AND METHODS: In vitro drug resistance of cells cultured from mammary carcinomas by the 3T3 feeder-layer technique was tested by the MTT assay. Out of the 33 tested cultures, 9 were derived from cells obtained from true-cut biopsies of primary tumours, with sample volume less than 0.03 cm3. The cultures were treated with 6 anticancer drugs at 6-8 concentrations. The chemoresistance of cultured cells was monitored by the surviving cell fraction as a function of the drug concentration. RESULTS: The average time to obtain a result was 28 days. The volume of an original sample had no effect on the in vitro resistance of a culture, suggesting minimal alteration of in vitro chemosensitivity of cells by their cultivation. Histopathological grade, estrogen receptor status or expression of the c-erb-B2 protein of the original tumours did not significantly correlate with the resistance of cultures. Individual drugs displayed distinct in vitro effectiveness. Paclitaxel and cisplatin were the most potent drugs. Gemcitabine, vinorelbine and mafosfamide were the least potent drugs. Doxorubicin and gemcitabine most frequently failed to completely metabolically inhibit 100% of cultured cells at any concentration. CONCLUSION: Combination of the optimised feeder-layer cultivation technique and the MTT test permits extensive drug resistance testing from very small breast cancer samples. PMID- 12894547 TI - Genomic instability in drug-resistant human melanoma cell lines detected by Alu-I arbitrary-primed PCR. AB - Destabilization of the genome seems to be an important step in the generation of drug resistance. Since malignant melanoma is extremely resistant to chemotherapy, we used human melanoma cell lines as a model to investigate the putative role of genomic instability in the appearance of drug resistance. Drug-resistant variants were obtained with MNNG, BiCNU, doxorubicin and 6-thioguanine selection of melanoma cell lines. Genomic alterations in variant cells were detected by arbitrarily primed PCR of Alu-I digested DNA (Alu-I-AP-PCR). Two differential DNA bands from 6-TG-resistant cell variants were sequenced. One is homologous to intron 25 of the neural cell adhesion molecule L1 and the second to endogenous retroviral LTR sequences. We have shown that drug-resistant melanoma cell lines accumulate genomic alterations that are efficiently detected by Alu I-AP-PCR and that drug-resistant variants show genomic instability, including variations in LTR sequences, which may be associated with the appearance of the drug resistance phenotype. PMID- 12894548 TI - Camptothecin conjugated with DNA minor-groove binder netropsin: enhanced lactone stability, inhibition of human DNA topoisomerase I and antiproliferative activity. AB - BACKGROUND: The conjugates of camptothecin (CPT) with ligands possessing different DNA selectivity could be promising agents in cancer therapy affecting expression of specific genes by trapping DNA topoisomerase I (top I)-DNA complexes in a sequence-selective manner. Our recent data show that minor-groove binder netropsin (Nt) and its derivatives modulate the CPT-induced pattern of top I-mediated DNA cleavage. In an effort to develop a new molecule with good biological activity we have linked CPT with Nt and report here the first results of in vitro examination of the new compound. MATERIALS AND METHODS: CPT-Nt conjugate linked with flexible spacer through position 7 of CPT chromophore was synthesized and analyzed for lactone stability, the ability to modulate a top I mediated DNA cleavage and antiproliferative activity within a panel of six tumor cell lines. RESULTS: CPT-Nt conjugate demonstrates enhanced lactone stability and concentration-dependent top I poisoning or suppression in vitro. The rate of conjugate hydrolysis in a water solution displays a 20-fold enhancement of stability compared with CPT. The cytotoxicity of the conjugate against acute promyelocytic leukaemia (HL60), chronic myelogenous leukaemia (K562), breast adenocarcinoma (MCF7), colorectal adenocarcinoma (HT29), lung carcinoma(A549) and ovarian adenocarcinoma (CaOV3) tumor cell lines was evaluated. The lowest IC50 value (0.08 microM) indicated its selective toxicity towards the ovarian adenocarcinoma cell line. CONCLUSION: The enhanced stability of CPT-Nt conjugate and its selective toxicity against the CaOV3 cell line may indicate its utility as an antitumor agent against ovarian adenocarcinoma. PMID- 12894549 TI - mRNA expression profile of matrix metalloproteinases and their tissue inhibitors in malignant and non-malignant prostatic tissue. AB - BACKGROUND: Increased expression of matrix metalloproteinases (MMPs) was found in various carcinomas. The aim of this study was to present a comprehensive mRNA expression profile of MMPs in benign and malignant prostatic tissue. MATERIALS AND METHODS: mRNA expression patterns of MMP-1, -2, -7, -9, -11, -14 and their tissue inhibitors (TIMPs) -1, -2 and -3 were studied in cancerous and non cancerous parts of 17 prostates removed by radical prostatectomy. Competitive reverse transcritpion PCR was used for quantification of MMP and TIMP mRNA. RESULTS: Both decreased (MMP-2, MMP-11, MMP-14) and a tendency to increased (MMP 9) MMP values in cancerous compared to the non-cancerous samples were observed. Significantly reduced TIMP-2 and TIMP-3 values were remarkable. Relatively strong associations were found among the three TIMPs while only a significant correlation between MMPs was observed between MMP-2 and MMP-7. There were no significant correlations between MMPs and tumor grade and stage and serum prostate-specific antigen. Receiver operation characteristic analyses proved that MMP and TIMP mRNA and their ratios have an insufficient capability to differentiate between cancerous and non-cancerous tissue. The ratios of MMP-9 to all three TIMPs and the ratio of MMP-14 to TIMP-3 were significantly increased. CONCLUSION: These increased ratios support the view of an imbalance between MMP-9 activity and its inhibitory counterparts in cancerous tissue as an important step in the development of prostate carcinoma and implicate the rationale of using synthetic inhibitors of MMPs as potential therapeutic tools. PMID- 12894550 TI - Murine hematopoietic stem cell and stromal responses to clinically-related, fractionated radiotherapy (FxRT). AB - The response of the blood-forming and supportive stromal compartments of the marrow to a localized, fractionated course of radiotherapy [FxRT; 2.0 Gy (q24 h x 5)-->74 Gy] was examined in the C57Bl/6 mouse using the hematopoietic progenitor and the cobblestone-forming cell (CAFC) assays as end-points for the blood forming compartment and the long-term bone marrow culture (LTBMC) to assess stromal integrity. Both during and subsequent to FxRT, hematopoietic activity in the irradiated femur was significantly dampened, although an abortive attempt at recovery was observed subsequent to the completion of FxRT. Moreover, both the CAFC subpopulations as well as the functional integrity of LTBMC generated from the irradiated femur were significantly compromised by FxRT. Of interest, restoration of the more primitive CAFC subpopulations to near normal levels was observed in the absence of a comparable recovery of the more mature CAFC subpopulations and the restoration of stromal integrity to the irradiated marrow. As a result, the data are consistent with the generation of a persisting FxRT induced lesion in the microenvironmental stroma that effectively interferes with the normal regulation of hematopoietic differentiation and the recovery of hematopoietic activity in the irradiated marrow. PMID- 12894551 TI - Application of the transgenic adenocarcinoma mouse prostate (TRAMP) model for pre clinical therapeutic studies. AB - BACKGROUND: The suitability of (C57BL/6 TRAMP x C57BL/6)F1 transgenic (TRAMP+) mice with well- to moderately-differentiated prostate cancer (PCa) was assessed for pre-clinical therapeutic studies. MATERIALS AND METHODS: TRAMP+ and TRAMP- mice were assessed for variability in genitourinary tract weight, seminal vesicle weight, prostate weight/volume and histopathology. Time-points included the reported ages of average tumour onset (approximately 25 weeks) and PCa-induced death (approximately 33 weeks). RESULTS: Seventy % of TRAMP+ mice aged 25-33 weeks had well- to moderately-differentiated PCa. At 25-28 weeks, the mean genitourinary tract weight was 2X greater and the mean prostate weight/volume was 1.5X more in TRAMP+ than in TRAMP- mice, respectively. Prostate weight/volume showed significant increases (p < 0.0001) by 2X and 3X, respectively by 31-33 weeks of age. CONCLUSION: The window for using the TRAMP model successfully for pre-clinical experimentation is 25-33 weeks provided that mice with poorly differentiated PCa showing a large tumour burden are excluded. PMID- 12894552 TI - Molecular mechanisms of hyperthermia- and cisplatin-induced cytotoxicity in T cell leukemia. AB - BACKGROUND: The prognosis of early and very early relapse in acute lymphoblastic leukemia of childhood is still very poor unless a hematopoietic stem cell transplant is performed if a second remission can be achieved by induction chemotherapy. Therefore an intensification of chemotherapy is required. MATERIALS AND METHODS: In the present study the molecular mechanisms of cisplatin- and/or hyperthermia-mediated cytotoxicity in CEM cells, a human T leukemia cell line, were investigated. RESULTS: Both hyperthermia and cisplatin induced the activation of the effector caspases-3 and -6. However, caspase activation followed different time kinetics. While hyperthermia exerted maximum caspase activation immediately after application, cisplatin activated caspase-3 and -6 after 24 hours. At both time-points significant caspase-3 and -6 activation was observed when the cells were stimulated by a combination of heat and cisplatin. The application of z-VAD-fmk, a general caspase inhibitor, showed that hyperthermia mediated cytotoxicity mainly via caspase-dependent mechanisms, while cisplatin induced both caspase-dependent and -independent cytotoxicity. Time kinetic experiments revealed that hyperthermia induced cell death immediately after the heating pulse. In contrast, cisplatin-induced cell death had its maximum between 6 hours and 12 hours after the heating pulse. The combined application of heat and cisplatin induced two peaks of cytotoxicity, one immediately after the heating pulse and the other between 6 hours and 12 hours. CONCLUSION: Hyperthermia and cisplatin induced cell death in T leukemic cells by different molecular mechanisms, which might explain the enhanced cisplatin induced cytotoxicity by hyperthermia. PMID- 12894553 TI - Silibinin induces growth inhibition and apoptotic cell death in human lung carcinoma cells. AB - BACKGROUND: The high systemic toxicity of chemotherapeutic agents limits their use to treat clinical lung cancer. These limitations could be minimized/overcome by using non-toxic phytochemicals, like, silibinin. MATERIALS AND METHODS: We used small cell lung carcinoma cells (SCLC) SHP-77 and non-small cell lung carcinoma cells (NSCLC) A-549, analyzing cell growth inhibition and death with Trypan blue exclusion, indices of the cell cycle progression with flow cytometry and apoptosis with propidium iodide and Hoechst 33342. RESULTS: Silibinin (25, 50 and 100 microM) treatment of SHP-77 and A-549 cells resulted in their growth inhibition and cell death. Cell cycle studies showed a small increase in G0-G1 population at all the time intervals in SHP-77 cells, however, in A-549 cells, a slight increase in G0-G1 but strong increase in S-phases was observed at lower treatment times, and a strong increase in G0-G1 population at 72 hours. Quantitative apoptotic studies showed that silibinin causes apoptotic cell death in both a dose- and a time-dependent manner with SHP-77 cells showing more apoptotic effect than A-549 cells. CONCLUSION: Silibinin significantly induces growth inhibition, a moderate cell cycle arrest and a strong apoptotic death in both small cell and non-small cell human lung carcinoma cells, which warrants further studies to assess the efficacy of this non-toxic agent in animal lung tumor models. PMID- 12894554 TI - Chemo- and radiation sensitivity of xenografted acute lymphoblastic leukemias- correlation to the expression of multidrug resistance proteins. AB - The aim of our study was to characterise, for the first time, the chemo- and radiation sensitivity of seven pediatric acute lymphoblastic leukemias xenotransplanted into immunodeficient NOD/SCID mice and to correlate the findings with the expression of three drug resistance proteins, P-glycoprotein (P-gp), multidrug resistance-associated protein (MRP1) and lung resistance protein (LRP). Mice were treated with single drugs used in clinical protocols: daunorubicin, doxorubicin, cyclophosphamide, vincristine, cytarabine, asparaginase and methotrexate. Two ALL samples, established from primarily diagnosed patients, responded to 5 or 6 of the tested cytostatics, respectively, while 3 out of 5 ALLs from relapse patients were only sensitive towards 2-4 drugs tested. Daunorubicin was more efficient than doxorubicin. The response of xenografted ALL toward vincristine and cyclophosphamide was inversely correlated with the expression of P-gp, LRP and MRP1 (R2 = 0.71, 0.70 and 0.64 for vincristine and 0.44, 0.70 and 0.60 for cyclophosphamide). A good correlation could be detected between the expression of P-gp and LRP (R2 = 0.88), P-gp and MRP1 (R2 = 0.75) and LRP and MRP1 (R2 = 0.90). The highest co-expression of the drug resistance proteins in the leukemia ALL-SCID 6 coincided with a high resistance to radiation and chemotherapy. Prediction of the individual drug resistance profile of a patient on the basis of results from the ALL-SCID xenograft studies was not possible because of the relatively long time necessary and because of the changes in the expression of P-gp, LRP and MRP1 during the murine generations. We conclude that in the drug resistance phenotype of ALL not only the above mentioned proteins but a variety of different molecules are involved. PMID- 12894555 TI - Induction of thymidine phosphorylase expression and enhancement of Furtulon sensitivity by Taxol in a human ovarian carcinoma cell line. AB - OBJECTIVE: Thymidine phosphorylase (TP) is one of the enzymes involved in the salvage pathways of nucleotide synthesis. The enzyme converts thymidine to thymine and 2'-deoxyribose-1-phosphate and can also metabolize the prodrug 5' deoxy-5-fluorouridine (Furtulon) to 5-fluorouracil and 5'-deoxy-D-ribose-1 phosphate. The aim of this study was to determine whether Paclitaxel (Taxol) induces TP expression and whether increased TP expression is further sensitized to Furtulon, using a human ovarian carcinoma cell line. MATERIAL AND METHODS: KF 28, a single-cell clone of a human ovarian carcinoma cell line, was used. TP expression was assessed by RT-PCR and Western blot analysis. Cell growth was evaluated by MTT assay. RESULTS: The concentration of Taxol that inhibited the growth of cells to the level of 50% of the control growth was 4.65 +/- 0.35 nM. TP gene expression was significantly increased at 72 hours 1 nM Taxol exposure compared to the control by RT-PCR (p = 0.020). Western blot analysis confirmed that the TP protein level was elevated compared to the control at 72 hours 1 nM Taxol exposure. Cell growth did not significantly differ between the control and 72 hours 1 nM Taxol exposure groups (p = 0.917). After 48 hours treatment with Furtulon followed by 72 hours 1 nM Taxol exposure, cell growth was dose dependently inhibited in Taxol-treated cells (p = 0.022), but not in non-Taxol treated cells (p = 0.082). CONCLUSION: Our results show that a low concentration of Taxol is a candidate for increasing TP expression in a human ovarian carcinoma cell line, and that cells with an elevated level of TP expression can be further sensitized to Furtulon. This information might be valuable in the development of new therapeutic interventions for epithelial ovarian cancer. PMID- 12894556 TI - HER-2/neu and CD117 (C-kit) overexpression in hepatocellular and pancreatic carcinoma. AB - BACKGROUND: Hepatocellular carcinoma has an overall 5-year survival of less than 5%. Similarly, pancreatic cancer has a mortality: incidence ratio of 0.99. The aim of this study was to determine the prevalence of HER-2/neu and c-kit (CD117) overexpression and to identify a possible predictive role in patients with these two malignancies. MATERIALS AND METHODS: We performed a retrospective study on archival specimens of subjects with hepatocellular and pancreatic carcinoma. HER 2/neu and CD117 overexpression were evaluated by immunohistochemistry. RESULTS: Thirty-three patients with pancreatic carcinoma and 25 patients with hepatocellular carcinoma were identified. The mean age was 71.7 years for patients with pancreatic carcinoma and 66 years for patients with hepatocellular carcinoma. Two patients with hepatocellular carcinoma and none with pancreatic cancer overexpressed HER-2/neu, while 2 patients with pancreatic carcinoma and 1 patient with hepatocellular carcinoma overexpressed CD117. CONCLUSION: HER-2/neu and CD117 are not significantly overexpressed in either cancer. There appears to be no role for the use of trastuzumab in either malignancy. Similarly, while there appears to be no role for tyrosine kinase inhibitors in the treatment of hepatocellular carcinoma, further larger studies are necessary in pancreatic adenocarcinoma. PMID- 12894557 TI - Flow cytometric evaluation of apoptosis and cell viability as a criterion of anti tumour drug toxicity. AB - BACKGROUND: Determination of the drug concentration required to kill 50% of the tumour cells (EC50) does not take into account the propensity of cells to undergo apoptosis and necrosis. These 2 parameters and the viable cells are here assessed by a flow cytometric (FC) approach using propidium iodide (PI) and FITC-Annexin V staining. MATERIALS AND METHODS: A number of carcinoma cell lines of defined p53 status were exposed to cis-PtII for 24 hours, stained with PI and FITC-Annexin V and analyzed by FC. Unstained viable cells, early apoptotic cells and necrotic cells were scored separately in dual parameter plots of green fluorescence (FITC) against red fluorescence (PI) to generate dose response curves. RESULTS: EC50 values for cell viability were found to be 1-4 times higher than survival data from colony assays resembling data obtained by MTT or Crystal Violet vital dye staining. Percentage apoptosis measured by Annexin V binding was in agreement with microscopic scoring of apoptotic cells after Acridine Orange staining. CONCLUSION: The FC assay described gives a good estimate of cell viability resembling data from vital dye staining assays and provides additional information on apoptosis and necrosis. FC data from Annexin V binding and microscopic scoring after Acridine Orange staining were in excellent agreement. PMID- 12894558 TI - Effect of MDR1 phosphorothioate antisense oligodeoxynucleotides in multidrug resistant human tumor cell lines and xenografts. AB - The effect of MDR1 antisense phosphorothiate oligodeoxynucleotides (S-ODNs) on resistant phenotype was investigated in multidrug-resistant human colon carcinoma and breast carcinoma cells in vitro and in vivo. Drug resistance in human colon carcinoma (SW620 Ad300) and breast carcinoma (MCF-7/INT500, MCF-7/AD150 and MCF 7/TH) cell lines is predominantly due to overexpression of P-glycoprotein (P-gp) resulting in decreased daunorubicin (DNR) accumulation. Two MDR1 antisense S ODNs, one complementary to the initial 15 bases of first exon (S-ODN I) and the other a loop forming sequence (S-ODN II) complementary to bases from 993-1007 of MDR1 gene, were tested for enhancing the doxorubicin (DOX) cytotoxicity in vitro and the efficiency of chemotherapy in human tumor xenografts. MDR1 antisense S ODN I reduced the DOX IC50 value 9-fold in multidrug-resistant SW620 Ad300 human colon carcinoma cells and 7 to 10-fold in breast carcinoma cells in vitro. The increase in DOX cytotoxicity correlated with a significant reduction of MDR1 mRNA in antisense S-ODN I-treated SW620 Ad300 cells. Even though the P-gp level was reduced at the end of the third day in antisense S-ODN I-treated cells, the rate of reduction was only partial compared to mRNA. The combination treatment of MDR1 antisense S-ODN I or II for three days and DOX for four days significantly controlled tumor growth rate in human tumors developed in nude mice. Our results suggest that MDR1 antisense S-ODN treatment can increase the efficiency of chemotherapy by suppressing gene expression and resistant phenotype. PMID- 12894559 TI - Evaluation of dHPLC in mutation screening of the APC gene in a Greek FAP cohort. AB - BACKGROUND: Germline mutations in the APC gene predispose to colorectal adenomas leading to cancer in over 80% of patients. A multitude of mutations, dispersed throughout the gene, have been described. We wanted to evaluate the usefulness of denaturing high performance liquid chromatography (dHPLC) for mutation screening. MATERIALS AND METHODS: Ten amplicons containing 14 mutations in the APC, previously identified by sequencing in 22 FAP patients, were analysed by dHPLC. dHPLC was also used to screen members of a family for a mutation identified in the proband. RESULTS: We analysed 10 amplicons under a total of 59 temperatures. Successful results were obtained from 51 out of 59 tested temperatures (86.4%). In all cases a different heteroduplex-homoduplex pattern was obtained from mutant DNA in at least two of the temperatures. All 14 mutations identified by sequencing were also detected using dHPLC. Sequence analysis of a large family confirmed the dHPLC results. CONCLUSION: Using dHPLC we detected all mutations previously identified by sequencing. PMID- 12894560 TI - Comparative morphometric analysis of primary versus recurrent basal cell carcinoma and of histological subtypes. Significance of morphometry of the nuclei. AB - BACKGROUND: Basal cell carcinomas (BCCs) are the most common cutaneous tumors, accounting for approximately 70% of all malignant skin tumors. A morphometric analysis was performed to evaluate whether morphometric parameters of the nuclei of BCC tumor cells may serve as predictors for biological behavior like recurrence rate or are able to differentiate morphological subtypes (multifocal superficial, nodular, sclerosing and miscellaneous forms of BCC). MATERIALS AND METHODS: N = 130 histological sections from n = 126 patients with BCC of the head and neck were analyzed. Morphometry included: maximum and minimum diameter, perimeter, nuclear area (direct parameters). According to the direct parameters, so called form factors were evaluated: axis ratio, shape factor, nuclear contour index, nuclear roundness and circumference/contour ratio. RESULTS: Morphometric parameters were not able to differentiate significantly between primary and recurrent BCC. Concerning the histological subtypes, multifocal superficial BCC differed significantly (< 0.05) from all other subtypes by direct parameters. CONCLUSION: Morphometry can help to distinguish between histological subtypes of BCC. Morphometry alone is not able to predict recurrence rate. PMID- 12894561 TI - Expression of cyclin-D1 in uveal malignant melanoma. AB - BACKGROUND: Uveal malignant melanoma (MM) is characterized by a marked variability in biological behavior and by an unpredictable clinical course. Therefore the search for reliable prognostic parameters is active. In this study we investigated the immunohistochemical expression of cyclin D1, a cell cycle regulatory molecule, to analyze the possible significance of the protein in the prognostic evaluation of these neoplasms. MATERIALS AND METHODS: Forty-five selected uveal MM were incubated with the anti-cyclin D-1 antibody; the findings were compared with the follow-up data, with the cell type and with the largest tumor dimension. RESULTS: The overexpression of cyclin D1 was found to be inversely related to disease-free interval (p < 0.001) and to the spindle A/B tumors with good prognosis (p = 0.05). On the other hand, a direct correlation was found with epithelioid/spindle B melanomas with poor clinical course (p = 0.05) and with sclera infiltration (p = 0.01). No statistically significant relation emerged between cyclin D1 expression and LTD. CONCLUSION: These data may suggest a possible role of the overexpression of cyclin D1 in the tumorigenesis of uveal MM and in the prognostic evaluation of these tumors, representing a useful tool to subclassify lesions with similar morphological features. PMID- 12894562 TI - CD7 expression in reactive and malignant human skin T-lymphocytes. AB - CD7, a molecule normally expressed on 90% of CD4+ T cells, is often deficient on the malignant T cells of cutaneous T cell lymphoma. Therefore deletion of CD7 is considered a specific marker for the diagnosis of cutaneous T cell lymphomas. Because an expansion of CD4+CD7- cells may also be observed in benign lymphocyte mediated dermatoses, we present our experience concerning CD7 expression in both cutaneous T cell lymphomas and a broad variety of T cell-mediated inflammatory dermatoses using an immunohistochemical approach on frozen sections from 45 patients. No, or at most scarce, expression of CD7 was detectable in the inflammatory skin conditions investigated as compared to CD3-positive cells. In cutaneous T cell lymphomas, a striking reduction of CD7 reactive cells was found in either reactive or malignant T cell components. Our findings indicate that CD7 negative T cells are more common within both benign and neoplastic T cell infiltrates than was previously demonstrated and suggest that, using a conventional immunoenzymatic technical approach on fresh-frozen sections, CD7 deletion is an unreliable criterion for the immunohistological diagnosis of T cell-mediated skin infiltrates. PMID- 12894563 TI - Detection of circulating tumor cells by cytokeratin 20 and prostate stem cell antigen RT-PCR in blood of patients with gastrointestinal cancers. AB - BACKGROUND: Detection of circulating tumor cells in blood may be an important diagnostic and prognostic factor in the management of tumor patients. The present study aimed to examine whether cytokeratin 20 (CK-20) and prostate stem cell antigen (PSCA) are useful markers for the detection of disseminated cancer cells in the blood of tumor patients. MATERIALS AND METHODS: A nested RT-PCR assay was used to detect CK-20 and PSCA mRNA in blood samples from 18 healthy donors, 15 patients with non-malignant disease, 9 patients with benign tumors and 47 patients with malignant tumors (11 pancreatic carcinoma, 8 gastric cancer, 15 colorectal carcinoma and 13 miscellaneous tumors). RESULTS: CK-20 expression was observed in the peripheral blood of 19 out of 47 (40.4%) patients with malignant tumors, 2 out of 9 (22.2%) patients with benign tumors and 3 out of 15 (20%) patients with non-tumor diseases. PSCA expression was present in the blood of 22 out of 47 (46.8%) patients with malignant tumors and particularly in 7 out of 11 (63.6%) patients with pancreatic cancer. CK-20 and PSCA expression was not observed in blood samples from healthy donors. There was a relationship between PSCA expression and tumor stage. CONCLUSION: The present results demonstrate that it is possible to apply a simple and reliable method for the detection of circulating tumor cells based on CK-20 and PSCA RT-PCR assays. PMID- 12894564 TI - PCD1, a novel gene containing PDZ and LIM domains, is overexpressed in human breast cancer and linked to lymph node metastasis. AB - BACKGROUND: Despite surgical removal of the primary tumor of breast cancer in patients with apparently localized disease, relapse at local or distant sites may occur because undetectable micrometastases were present at the time of diagnosis. Identification of molecules associated with breast cancer metastasis suggests possible new treatments. PCD1, a gene encoding a new member of the PDZ and LIM domain-containing protein family, recently was identified. We examined the relationships between PCD1 mRNA expression in breast cancers and metastasis. MATERIALS AND METHODS: PCD1 mRNA expression in breast cancer tissues was examined using a quantitative reverse transcription polymerase chain reaction. RESULTS: PCD1 mRNA expression was greater in cancers than in noncancerous tissues (p < 0.0001). In addition, high PCD1 gene expression was more frequent in patients with lymph node metastasis. CONCLUSION: PCD1 appears to contribute to breast cancer progression and nodal metastasis, thus representing a potential target molecule in breast cancer diagnosis and treatment. PMID- 12894565 TI - Combined radiation and cytochrome CYP4B1/4-ipomeanol gene therapy using the EGR1 promoter. AB - BACKGROUND: Cytochrome p450 isozyme CYP4B1 converts the inert prodrug 4-ipomeanol (4-IM) into toxic alkylating metabolites. Induction of cytotoxicity by 4-IM combined with ionizing radiation (IR) in cells transfected with a fusion protein of rabbit cytochrome CYP4B1 under control of the radiation inducible EGR1 promoter was investigated. The capability of activated 4-IM to sensitize cells to IR was also assessed. MATERIALS AND METHODS: Survival fractions of cells, determined by MTT assays, stably transfected with EGR1-CYP4B1 were compared with that of cells transfected with a control plasmid after IR followed by 4-IM. Radiosensitization was tested by comparing clonogenic survival curves of cells transfected with the CYP4B1 cassette under a CMV promoter instead of EGR-1, irradiated with or without 4-IM. RESULTS: MTT assays for cytotoxicity indicated a decrease in relative survival fractions (survival with 4-IM/survival without 4 IM) of the EGR1-CYP4B1 transfected cells with increasing radiation dosage, but not of control cells. Clonogenic assays revealed decreased survival fractions with increasing radiation doses (CYP4B1 transfected and control cells) and 4-IM concentrations (CYP4B1 transfected cells), but showed no significant differences in slope of survival curves with 4-IM. CONCLUSION: The results indicate IR potentiates the cytotoxic activity of the EGR1-CYP4B1/4-IM transgene system, but activated 4-IM does not sensitize cells to IR. Thus, the EGR1-CYP4B1/4-IM system is a viable radiation-gene therapy system that may allow for improved spatial and temporal control of cytotoxicity by therapeutic radiation fields. PMID- 12894566 TI - Prognostic significance of hepatoma-up-regulated protein expression in patients with urinary bladder transitional cell carcinoma. AB - We recently found a urinary molecular marker, Hepatoma-Up-Regulated Protein (HURP), for the detection of urinary bladder transitional cell carcinoma (TCC). In this study, the expression of HURP in 57 voided urine specimens was determined by semi-quantitative reverse transcriptase-polymerase chain reaction. HURP value was correlated with a variety of clinical parameters, including sex, age, tumor grade, stage, multiplicity, tumor shape and recurrence/metastasis during the follow-up period. The detection rate of HURP was 60.7% (17 out of 28) in voided urine of patients with TCC, 5.9% (1 out of 17) in non-TCC urological benign disease and 0% (0 out of 12) in healthy volunteers. HURP positivity (defined as above 0.1 cut-off value) was detected in 83.6% (92 out of 110) TCC specimens, 4.3% (1 out of 23) of non-TCC urinary cancer and 0% (0 out of 15) of benign urological disease. No relationship was found between the level of HURP and the above-mentioned clinical parameters other than recurrence of TCC patients. A higher level of tissue HURP was found in the patients having recurrence (Mann Whitney U-test, p = 0.027). PMID- 12894567 TI - MDR1 single nucleotide polymorphism C3435T in normal colorectal tissue and colorectal carcinomas detected by MALDI-TOF mass spectrometry. AB - Single nucleotide polymorphisms (SNPs) may contribute to the malignant process and may show clinicopathological importance as prognostic markers. The multidrug resistance gene MDR1 encodes a membrane transporter which confers cytostatic drug resistance in tumors and protects normal tissues from xenobiotics. We analyzed the C3435T SNP in the MDR1 gene which is associated with altered cellular drug uptake in matched tumor and normal tissues of 45 patients suffering from colorectal carcinoma. We have developed a highly sensitive matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) method to survey the C3435T polymorphism in PCR-amplified fragments of the MDR1 gene. Thirteen patients were homozygous for C/C (29%), 15 were heterozygous (33%) and 17 were homozygous for T/T (38%). None of the tumor samples showed an altered SNP compared to their matched normal tissue samples. As analyzed by the Kruskall Wallis test, none of the clinicopathological parameters was significantly associated with homo- or heterozygosity. The combination of PCR, allele-specific primer extension reactions and MALDI-TOF-MS offers a promising alternative method for genotyping the MDR1 gene especially for heterozygous situations. The inherent advantages of MALDI-TOF-MS based genotyping include its high molecular resolution, high signal-to-noise-ratios and reproducibility, combined with an excellent sensitivity. As none of the tumor samples showed an altered state compared to their matched normal tissue samples, the genotypic frequency of this polymorphism seems not to be altered during colorectal tumorigenesis. PMID- 12894568 TI - The effect of rifampin on the pharmacokinetics of vinorelbine in the micropig. AB - BACKGROUND: Vinorelbine has been shown to be metabolised by CYP3A4 in vitro. To evaluate the impact of CYP3A in the disposition of vinorelbine in vivo, we compared the kinetics of the alkaloid given intravenously alone and combined with rifampin, a potent CYP3A inducer, in the micropig. ANIMALS AND METHODS: Four healthy Yucatan micropigs, about 20 kg, received a first infusion of vinorelbine (0.5 mg/kg). During the next week they were injected rifampin (600 mg daily) and a second vinorelbine infusion (0.5 mg/kg) on the 7th day of rifampin dosing. Serum concentrations of vinorelbine and rifampin were measured by high performance liquid chromatography. RESULTS: The mean peak concentrations of vinorelbine were 274.2 ng/ml (Standard Deviation or SD: 90) and 458 ng/ml (SD: 448), the mean areas under the serum concentration-time curve were 8,344 ng.min.ml-1 (SD: 2,604) and 14,093 ng/ml.min-1 (SD: 10,000) and the total clearances were 1.146 l/min (SD: 0.333) and 1.003 l/min (SD: 0.714) when the Catharanthus alkaloid was given alone or was combined with rifampin, respectively. CONCLUSION: We did not observe an increase in vinorelbine elimination by rifampin related to a CYP3A induction in an animal model physiologically close to humans. Although the number of animals was small, these results suggest that CYP3A metabolism constitutes a minor pathway of elimination of intravenous vinorelbine in the micropig. PMID- 12894569 TI - Synthesis, radiolabelling and biological characteristics of a bombesin peptide analog as a tumor imaging agent. AB - BACKGROUND: Several human cancers, including small cell lung, prostate, breast, gastric, colon and pancreatic cancers, express receptors for bombesin-like peptides. Bombesin (BN) peptides that bind specifically to these receptors are useful for detection of bombesin receptor-expressing cancers in vivo. A new 99mTc labelled-BN peptide for targeting bombesin receptor-expressing cancers was prepared and characterized. MATERIALS AND METHODS: MAG3-coupled BN peptide (MAG3 BN) was prepared by solid-phase synthesis and radiolabelled with 99mTc by an exchange method. In vitro cell binding assays were conducted on human breast cancer cell lines, MDA-MB-231 and MCF-7. In vivo biodistribution studies were performed in normal and nude mice bearing bombesin receptor-positive tumors. RESULTS: Radiolabelling of MAG3-BN with 99mTc produces a single radioactive species (> 95%). In vitro cell-binding indicated the affinity and specificity of 99mTc-MAG3-BN towards bombesin receptors. In vivo biodistribution in mice demonstrated that 99mTc-MAG3-BN cleared rapidly from the blood and most non targeted tissues and was excreted mainly via the kidneys. Uptake in bombesin receptor-positive tissues and in the tumor was low to moderate. CONCLUSION: 99mTc MAG3-BN displays good radiolabelling together with certain favorable biological characteristics and might be a useful peptide radiopharmaceutical in the detection of bombesin receptor-expressing cancers in vivo. PMID- 12894570 TI - Fungal beta-(1-3)-D-glucan derivatives exhibit high antioxidative and antimutagenic activity in vitro. AB - The antioxidative activity and antimutagenic effects of the water-soluble beta-(1 3)-D-glucan derivatives from biotechnologically important species, in particular carboxymethyl-glucan (CM-G) and sulfoethyl-glucan (SE-G) both from the baker's yeast Saccharomyces cerevisiae, and carboxymethyl-chitin-glucan (CM-CG) from filamentous fungus Aspergillus niger, were evaluated. The luminol-dependent photochemical method using trolox as a standard showed that CM-CG, SE-G and CM-G possessed high antioxidative properties. CM-CG exhibited the highest antioxidative activity (2.15 +/- 0.14 nmol exhibits the same activity as 1 nmol of trolox), followed by SE-G (2.99 +/- 0.15 nmol) and CM-G (4.59 +/- 0.14 nmol). These glucans were experimentally confirmed to exhibit different, statistically significant activity in reducing the damage of chloroplast DNA of the flagellate Euglena gracilis induced by ofloxacin and acridine orange. Our findings suggest that the antimutagenic effect of CM-CG, SE-G and CM-G against ofloxacin is based on their antioxidative capability to scavenge reactive oxygen species (p < 0.001). As far as acridine orange is concerned, the reduction of the chloroplast DNA lesion could be a result of the absorptive capacity of the glucans (p < 0.001). We found out that the water-soluble beta-(1-3)-D-glucan derivatives possess very high antioxidative activity as well as expressive antimutagenic effects, exerted through different mode of action. PMID- 12894571 TI - Immune responses to human tumors: development of tumor vaccines. AB - Strong evidence has been accumulated demonstrating that tumor cells in humans and animal are recognized in general as non-self by the immune system and they are able to induce an immune response which often leads to their elimination. In humans, this evidence includes: (a) The development of T-cell lines and clones with antitumor activity (cytotoxic or helper) which is restricted to autologous tumor cells or to cells expressing the same tumor peptide/HLA epitope; (b) the presence of oligoclonal T cells infiltrating many tumors; (c) the identification and molecular cloning of tumor antigens and of peptides derived from these antigens, which elicit HLA-restricted immune responses. Their discovery provided the ultimate proof for the presence of specific immune responses in human tumors. The availability for the first time of molecularly cloned tumor antigens permitted the development of peptide or recombinant tumor vaccines. Although significant progress has been made and tumor peptide vaccines capable of eliciting biological responses in more than 50% of the patients and objective clinical responses in 10 to 42% of the patients have been reported, certain major problems remain and need to be resolved in order to develop effective tumor vaccines. These problems emanate from the following mechanisms that the tumor cells are employing to avoid detection and destruction by the immune system: (i) Down-regulation of HLA class I expression on the surface of tumor cells; (ii) Down-regulation of tumor antigen expression or selection of negative tumor variants; (iii) Expression of naturally occurring altered peptide ligands by tumor cells; (iv) Lack of costimulatory molecules on tumors cells; (v) Production of immunosuppressive cytokines, such as TGF-beta and IL-10; (vi) Induction of lymphocyte apoptosis by tumor cells using the Fas/Fas L pathway; (vii) Down regulation or absence of CD3 zeta (zeta) transcripts or protein in tumor infiltrating lymphocytes (TIL), and others. The selection of optimal tumor antigens for vaccine development is another issue that requires attention. Lineage specific or differentiation antigens appear to be better candidates for the development of tumor vaccines because they are expressed in all tumor cells. Methods for antigen presentation, such as those using dendritic cells, also play a critical role in the development of tumor vaccines. In addition to the progress towards the development of tumor vaccines, substantial progress has been made in developing advanced methods of adoptive immunotherapy based on TIL. This approach can be effective when an immune response can not be elicited in vivo. The progress made towards the development of tumor vaccines and approaches for adoptive immunotherapy has been substantial. Additional studies need to be carried out to develop new and effective tumor vaccines and adoptive immunotherapy methods. PMID- 12894572 TI - siRNAs, ribozymes and RNA decoys in modeling stem cell-based gene therapy for HIV/AIDS. AB - Gene therapy strategies for HIV infection require gene transduction of hematopoietic stem cells with effective therapeutic constructs. Here we summarize our studies on anti-HIV ribozymes, RNA decoys and the newly described siRNAs. The therapeutic constructs consisted of an anti-CCR5 ribozyme to down-regulate the HIV-1 cell surface co-receptor and ribozymes targeted to viral mRNAs coding for the tat, rev and env proteins. The RNA decoy targeted rev and the siRNA was directed against a sequence common to rev and tat mRNAs. CD34 hematopoietic progenitor cells were transduced with retroviral or lentiviral vectors containing these constructs. They were differentiated into macrophages in vitro and T cells in vivo in a SCID-hu mouse thymopoiesis model. The transgene-containing macrophages and T cells were found to be phenotypically normal. When challenged in vitro with HIV-1, they showed significant anti-viral resistance. These proof of-concept studies demonstrated the utility of RNA-based anti-HIV constructs for gene therapy. PMID- 12894573 TI - CaSm antisense gene therapy: a novel approach for the treatment of pancreatic cancer. AB - Pancreatic adenocarcinoma is a major clinical problem with few effective treatment options. In the United States 29,000 cases are diagnosed annually with an associated mortality rate greater than 90%. Given this dismal prognosis, a better understanding of the molecular controls that govern pancreatic cancer is clearly needed in order to develop more effective therapies. As such, our group has been actively investigating the identification and potential application of novel gene targets for this disease. We have recently identified the cancer associated Sm-like (CaSm) oncogene, shown that it is overexpressed in 87% of human pancreatic cancer samples, and clearly demonstrated that it functions as a classic oncogene. We have also been able to show that an adenovirus expressing antisense RNA to the CaSm gene (Ad-alpha CaSm) is able to reduce endogenous CaSm mRNA expression and decrease anchorage-independent growth. A single intratumor injection of Ad-alpha CaSm extended survival in an in vivo SCID mouse model of human pancreatic cancer. To gain insight into the mechanism of Ad-alpha CaSm's anti-tumor effect, cell cycle studies were performed. Ad-alpha CaSm treatment of pancreatic cancer cells resulted in a cytostatic block with decreased G1-phase and increased DNA content in vitro. Importantly, the combination of Ad-alpha CaSm with gemcitabine (an S-phase active chemotherapy) significantly extended survival time beyond either therapy alone. These studies have defined the CaSm oncogene as a novel gene target for therapy and have begun to define its potential role in the pathogenesis of pancreatic cancer. PMID- 12894574 TI - Marker-assisted study of genetic background and gene-targeted locus modifiers in lymphopoietic phenotypes. AB - Forward and reverse genetic approaches facilitate the molecular dissection of individual gene functions and the integration of individual gene functions into multi-gene processes in the context of the whole organism. Variations in mutant phenotypes due to genetic background differences have been well documented through the analysis of mouse mutants. Nevertheless, recommendations concerning the assessment of genetic background as it impacts on phenotype, and utilization of genetic background differences to identify and integrate gene functions have been largely overlooked. Genetic background as it relates to immunological mutants will be discussed utilizing an Ets1-targeted allele to exemplify phenotypic variation due to background. Marker-assisted strategies for the identification of genetic modifiers, especially those linked to the targeted locus, will also be considered. PMID- 12894575 TI - The breast cancer-associated gene Di12 has oncogenic activity. AB - The breast cancer-associated gene Di12 encodes a novel protein, which was found overexpressed in invasive ductal carcinomas of the breast. In experiments designed to assess the role of the Di12 gene in oncogenesis, the overexpression of 339 N-terminal amino acids of this gene in NIH3T3 cells resulted in cellular transformation and in vivo tumorigenesis. NIH3T3-Di12 tumor cell growth was partly reversible upon Di12 antisense treatment. In addition, transformation of the ER+ human breast cancer cell line MCF-7 resulted in hormone independent growth of these tumors in nude mice. Di12 expression in NIH3T3 and MCF-7 tumor cells was confirmed by RT-PCR and mabDi12 immunostaining. Immunohistochemistry using mabDi12 on an arrayed collection of 106 invasive breast tumors further underlined the expression of the gene in over 75% of advanced stage breast cancers. Our data indicate that Di12 expression is oncogenic in in vitro transformation and in vivo tumorigenic assays. PMID- 12894576 TI - T-antigen of human polyomavirus JC cooperates withIGF-IR signaling system in cerebellar tumors of the childhood-medulloblastomas. AB - Polyomaviruses are implicated in a number of cancers, and the transforming activity of their early protein, large T-antigen, has been documented in a variety of cell types and in experimental animals (1). Although the pathways by which T-antigen induces uncontrolled cell growth are not fully defined, T-antigen mediated inactivation of tumor suppressors, p53 and pRB, is well-documented in some malignancies (2). Here we postulate that functional interaction between the insulin-like growth factor (IGF-IR) and the T-antigen of human polyomavirus JC (JCV T-antigen) may contribute to the process of malignant transformation in medulloblastomas: (i) the IGF-IR signaling system is strongly activated in medulloblastoma cell lines and medulloblastoma biopsies; (ii) the cytoplasmic protein, insulin receptor substrate 1 (IRS-1), is translocated to the nucleus in the presence of JCV T-antigen; (iii) molecular characterization of the interaction between IRS-1 and JCV T-antigen indicates that the binding involves the N-terminal portion of IRS-1 (PH/PTB domain) and the C-terminal region of JCV T-antigen (aa 411-628); and finally (iv) competition for the IRS-1-JCV T-antigen binding attenuates anchorage-independent growth of T-antigen positive medulloblastoma cells in culture. Based on these findings, we propose a novel role for IRS-1 in JCV T-antigen-mediated deregulation of cellular equilibrium, which may involve uncoupling of IRS-1 from the surface receptor and translocation of its function to the nuclear compartment of the cell. PMID- 12894577 TI - Gene expression profiling of ductal carcinomas in situ and invasive breast tumors. AB - Comparative and functional genomics are powerful tools to advance the understanding of the molecular basis of cancer. It is believed that genes are epigenetically regulated and, thus, each tumor type and stage will be characterized by a gene expression fingerprint. In this study we identified genes that are differentially expressed in ductal carcinoma in situ and invasive ductal carcinoma of the breast. To isolate genes that are associated with progression of breast cancer we performed differential display and subtractive cloning procedures using matched RNA from normal and tumor tissue. cDNA microarray analysis generated gene expression profiles typical of the transition from in situ to invasive breast cancer when we used mRNA extracted from a case of low- to intermediate-grade DCIS and a case of high-grade DCIS/IDC. cDNAs from these samples were the probes in a cDNA microarray hybridization to 9183 unique cDNAs representing 8507 genes. Signals from both transcriptomes were obtained for 8083 genes, and the balanced differential expression values between pure DCIS and DCIS/invasive tumors revealed 303 distinct cDNAs with a ratio of > 2. Interferon inducible genes were found to be expressed at the highest level in the pure DCIS sample. Genes most abundantly expressed in the invasive tumor were immunoglobulin heavy constant gamma 3 and calgranulin B. Further analysis of RNA and protein expression in breast tumor cell lines and patient tissue samples revealed that: IGFBP-rP1 is down-regulated in invasive tumors whereas cyclin I protein is regulated by ubiquitination and is associated with ER-negative breast cancers. CONCLUSION: The known and novel genes discussed here represent targets for molecular characterization during breast cancer development as well as for designing novel strategies for diagnosis and treatment. PMID- 12894578 TI - C-erbB2, p27 and G1/S aberrations in human primary breast cancer. AB - BACKGROUND: C-erbB2 is overexpressed in approximately one-fourth of human breast cancers. In spite of numerous reports suggesting a connection of c-erbB2 overexpression with cell cycle regulation through p27, D cyclins and c-myc, the relationship between c-erbB2 and proliferation through de-regulation of the pRb pathway in primary breast cancer has not been fully clarified. MATERIALS AND METHODS: For this purpose we compared the expression of c-erbB2 in a total of 105 primary breast tumours with a variety of cell cycle proteins and clinical parameters. RESULTS: C-erbB2 was strongly correlated with down-regulation of p27 and overexpression of c-erbB2 was, as expected, associated with poor survival. However, there was no correlation with proliferation. There was, nevertheless, an association between c-erbB2 and proliferation in certain subtypes of breast cancer, as in oestrogen-receptor-positive tumours, tumours with high cyclin D1, and in tumours with an overall linear pRb pathway, i.e. tumours with a preserved linearity between cyclin D1, pRb phosphorylation and proliferation. CONCLUSION: Our results suggest that c-erbB2 may have alternative functions in different subtypes of breast cancer, and further stress that c-erbB2, in addition to promoting proliferation, also functions through other mechanisms in breast cancer. PMID- 12894579 TI - Varicella zoster virus transcription in latently-infected human ganglia. AB - Varicella zoster virus (VZV) causes childhood chickenpox, becomes latent in cranial, dorsal root and autonomic ganglia, and can reactivate decades later to cause shingles and other serious neurological complications. Herein, we summarize investigations conducted over the past decade that have identified virus genes expressed in latently-infected human ganglia. A model of VZV gene regulation during latent infection was tested and future directions in the study of VZV latency are discussed. PMID- 12894580 TI - Natural killer cells are persistently infected and resistant to direct killing by HIV-1. AB - We have reported the presence of a CD4+ subpopulation of Natural Killer cells (NK) in humans. These cells also express HIV coreceptors CCR5 or CXCR4 and are infectable by HIV-1. Infection is productive and persistent in vitro and in vivo. Infected people under therapy have evidence of viral DNA in their NK pool. In vitro experiments show that the NK compartment is important for virus propagation, since HIV-1 growth is significantly reduced in PBMC depleted of NK cells. This reduction was observed for both R5 and X4 molecular clones of HIV-1. In addition, evidence is presented demonstrating that infected NK cells are killed less efficiently by the virus as compared to T cells. This is reminiscent of macrophage infection, where the virus-infected cells survive for long periods of time. Therefore, few infected NK cells may contribute significantly to the infectious virus produced in vitro. These results further underscore the importance of NK infection for understanding pathogenic mechanisms leading to AIDS, as well as the various reservoirs and sanctuaries for HIV upon antiretroviral treatment. PMID- 12894581 TI - Targeting the A3 adenosine receptor for cancer therapy: inhibition of prostate carcinoma cell growth by A3AR agonist. AB - BACKGROUND: Agonists to A3 adenosine receptor (A3AR) were shown to inhibit the growth of various tumor cell types. The present study demonstrates that a synthetic A3AR agonist, 1-deoxy-1-[6-[[(3-iodophenyl)methyl]amino]-9H-purine- 9 yl]-N-methyl-beta-D-ribofura-nuronamide (IB-MECA), inhibits the growth of androgen-independent PC-3 prostate human carcinoma cells and illustrates the molecular mechanism involved. MATERIALS AND METHODS: PC-3 prostate carcinoma cells were used. Cell growth was examined in vitro by the thymidine incorporation assay and in vivo by inoculating the tumor cells subcutaneously into nude mice and monitoring tumor size. The protein expression level in cells and tumor extracts was tested by Western blot analysis. RESULTS: A decrease in the protein expression level of A3AR and the downstream effector PKAc was observed. Consequently, the GSK-3 beta protein level increased, resulting in the destabilization of beta-catenin and the subsequent suppression of cyclin D1 and c myc expression. IB-MECA treatment also induced down-modulation of the expression of NF-kappa B/p65, known to regulate the transcription of cyclin D1 and c-Myc. This chain of events occurred both in vitro and in vivo and suggests the use of the above-mentioned signaling proteins as markers to predict tumor cell response to A3AR activation. CONCLUSION: Taken together, we demonstrated that A3AR activation deregulates the Wnt and the NF-kappa B signaling pathways resulting in the inhibition of prostate carcinoma cell growth. PMID- 12894582 TI - ARF1 and ARF6 are dispensable for Crk-dependent epithelial-mesenchymal-like transitions. AB - BACKGROUND: Activation of the Met receptor tyrosine kinase through its ligand, hepatocyte growth factor (HGF), promotes an epithelial-mesenchymal transition and cell dispersal. However, little is known about the HGF-dependent signals that regulate these events. HGF stimulation of epithelial cell colonies leads to the enhanced recruitment of the CrkII and CrkL adapter proteins to Met-dependent signaling complexes. Overexpression of Crk adapter proteins in MDCK cells promotes spreading and loss of adherens junctions, events regulated by HGF. We recently demonstrated that the overexpression of CrkII promotes the formation of a multi-molecular complex containing CrkII, Paxillin and GIT-2, an ARF-GAP. MATERIALS AND METHODS: To determine the possible role of ARF1 and ARF6 in Crk- and HGF-dependent cell spreading, dominant negative mutants of ARF1 or ARF6 were microinjected into MDCK cells. RESULTS: We report that MDCK cell lines overexpressing CrkII display reduced ARF6 but not ARF1 activity. While both ARF1 and ARF6 are required for the spreading of MDCK cells stimulated with HGF, ARF1 and ARF6 activity is dispensable for the spreading of cells microinjected with Crk. CONCLUSION: We propose that Crk adapter proteins may act downstream of ARF1 and ARF6 to promote cell spreading or rely on different pathways to enhance cell spreading. PMID- 12894583 TI - The Pur protein family: clues to function from recent studies on cancer and AIDS. AB - The Pur protein family consists of four known members in humans, the prototype gene for which is strongly conserved throughout evolution. Several investigations have now implicated Pur alpha in pathways of inhibition of oncogenic transformation. Clues to the potential importance of Pur family members in cancer are derived from observations of genetic alterations of both Pur alpha and Pur beta in myelodysplastic syndrome progressing to acute myelogenous leukemia. A role for newly-discovered Pur gamma in neoplasia is also beginning to emerge as studies have indicated that cellular levels of two Pur gamma isoforms are elevated in certain tumors. A variety of studies have now implicated Pur alpha in development of blood cells and cells of the central nervous system. Clues to the functions of Pur alpha, a key family member, have recently been derived from studies of the interactions of HIV-1 and JC virus (JCV) in AIDS. JCV causes an opportunistic infection in the brains of certain HIV-1-infected individuals. Pur alpha can influence this viral interaction through functional associations with the Tat protein and TAR RNA of HIV-1, and with large T-antigen and DNA regulatory regions of JCV. Evidence is now strong that Pur alpha interacts with both DNA and RNA and that an important aspect of its function is to recruit regulatory proteins to specific nucleic acid sequences in processes as diverse as DNA replication, gene transcription, RNA transport in the cytoplasm and compartmentalized mRNA translation. PMID- 12894584 TI - The study of HOX gene function in hematopoietic, breast and lung carcinogenesis. AB - HOX transcription factors regulate basic and cell type-specific activities throughout life. The combinatorial patterns of HOX gene expression along anterior posterior and proximal-distal axes are relatively tightly-defined during embryogenesis and key remnants of such positional memory persist through adulthood. These normal patterns of HOX gene expression can be compared to a growing body of work on their dysregulation during carcinogenesis. In this review, simple and complex changes in HOX gene expression patterns will be considered using examples from hematopoietic, breast and lung cancers. Changes in individual and combinatorial patterns of HOX gene expression, co-factor expression and chromatin structure will be considered in a discussion of potential roles for dysregulated HOX genes in target gene regulation and various aspects of cancer progression. Collectively, studies indicate that, although a variety of factors must be delineated to assess the roles of individual HOX genes in particular cancers, approaches that modulate HOX gene expression and monitor both changes in the regulation of key target genes and cellular activities are making the greatest initial advances in this assessment. PMID- 12894585 TI - Glutathione S-transferase M1 polymorphism and the risk of lung cancer. AB - Glutathione S-transferases (GSTs) are a family of enzymes that detoxify hydrophobic electrophiles, including polycyclic aromatic hydrocarbon carcinogens that have been implicated in the pathogenesis of lung cancer. The GSTM1 gene within the mu class of human GSTs has been shown to be polymorphic, with individuals who are homozygous for a null allele having the GSTM1-null genotype. Individuals with the GSTM1-null genotype are deficient in both the GSTM1 and GSTM3 isoenzymes in the lung. A number of epidemiological studies have been conducted to assess the association between the GSTM1-null genotype and the risk of lung cancer. Although there have been conflicting reports regarding this relationship, the current weight of evidence indicates that the GSTM1-null genotype is probably associated with a modest increase in the risk of lung cancer. This risk appears to be greater in African-American and Asian populations than in Caucasians. Recent investigations have shown that the GSTM1-null genotype combined with CYP1A1, NAT2, or GSTP1 polymorphisms confers a greater risk of lung cancer than the GSTM1-null genotype alone. Future investigations should focus on assessing the risk of lung cancer related to multiple combinations of genetic polymorphisms that may identify individuals who are at high risk for developing lung cancer with a greater degree of certainty than is currently possible. This could lead to new clinical strategies for counseling, risk reduction and the detection of lung at an early and potentially curable stage. PMID- 12894586 TI - The epithelial-specific Ets factors occupy a unique position in defining epithelial proliferation, differentiation and carcinogenesis. AB - The Ets family of transcription factors regulates many biological processes. Within the Ets family are a subset of proteins that have epithelial restricted expression patterns, both in tissues and cell lines. These Epithelial-specific Ets (Ese) factors, cluster into two groups based on the sequence of the Ets DNA binding domain: (i) Ese1, Ese2 and Ese3, and (ii) the more divergent, Pdef. Gene targeting and cell culture studies have demonstrated that Ese factors are important for cellular differentiation. Furthermore, the ability of Ese factors to regulate expression of genes that are markers of differentiation supports this model. Significantly, the expression profile of these Ese factors is altered in epithelial cancers relative to the respective normal tissues. It is likely that this variation in expression affects downstream target genes important for carcinogenesis such as oncogenes and tumor suppressor genes. Understanding the role of these Ese factors in normal cellular differentiation may elucidate specific mechanisms by which cancer cells can become undifferentiated. PMID- 12894587 TI - Epstein-Barr virus induces invasion and metastasis factors. AB - The oncogenic properties of the principal EBV oncoprotein, Latent Membrane Protein 1 (LMP-1), include the ability to induce invasiveness and metastasis factors. We have shown that LMP-1 induces matrix metalloproteinase 9 (MMP-9), a type IV collagenase that disrupts basement membrane. Also, cyclooxygenase-2 (COX 2), which is overexpressed in diverse malignancies, is induced by LMP-1; the enzyme is functional, and co-expressed with LMP-1 in NPC. Inhibitors of the NF kappa B signaling pathway, which is activated by LMP-1, including I kappa B super repressor and aspirin reduce or cancel induction of MMP-9, COX-2 and invasiveness of LMP-1-expressing cells. Production of VEGF, also induced by LMP-1, is decreased by a COX-2-specific inhibitor. We now show that LMP-1 induces expression of the angiogenic Fibroblast Growth Factor-2 (FGF-2). Furthermore, LMP 1 also causes secretion of the 18 kDa isoform of this protein--a newly identified function for LMP-1. Secretion of FGF-2 is independently signaled through the NF kappa B pathway. Release of the protein is not dependent on the classical ER/Golgi secretory pathway, but secretion of FGF-2 is suppressed by ouabain, an inhibitor of the Na+/K(+)-ATPase alpha 1 subunit. Finally LMP-1 induces expression of Hypoxia-Inducible Factor-1 alpha (HIF-1 alpha), which mediates adaptation of cells to O2-depleted states. Thus LMP-1 is not only directly oncogenic, it can induce a constellation of factors that reveal the additional role of EBV in invasive cancers such as NPC. Alteration of cellular phenotype independent of transforming effects may be a property of other tumor viruses. PMID- 12894588 TI - 99MTC [13LEU] bombesin and a new gamma camera, the imaging probe, are able to guide mammotome breast biopsy. AB - A pilot study has been carried out in order to verify the feasibility of scintigraphic driving for breast biopsy. A new high resolution (HR) gamma ray detector, the imaging probe (IP), and 99mTc [13Leu] Bombesin (99mTc BN), have been used to drive a mammotome biopsy needle after having fused radioisotope with digital X-ray images. IP is a mobile, high resolution, miniaturised gamma camera, whose field of view is one inch and whose spatial resolution is 2 mm. 99mTc BN is a new radiotracer derived from the well-known peptide, that has already shown very high sensitivity in detecting breast cancer. PATIENTS AND METHODS: Five patients very suspicious for breast cancer were studied. 185 MBq of 99mTc BN were i.v. injected and dynamic prone scintimammography was performed for 20 minutes with a conventional large field of view gamma camera. IP was matched with the biopsy system and digital X-ray device of a mammotome system, in order to fuse images and to use the mammotome pointer indifferently on X-ray, scintigraphic and fused images. Biopsy samples were counted and weighed: uptake was expressed as counts sec-1 gr-1. Samples were classified into high, intermediate and low uptake. Conventional histological assessment was blindly performed on the samples. RESULTS: All of the patients showed cancer. The T categorisation was T1a for two cancers and T1b for 3. Dynamic prone scintimammography with conventional gamma camera, as well as HR scintigraphy with IP showed spots of 99mTc BN uptake. Maximal mismatch between X-ray lesions and of hot spots imaged with IP before mammotome scintigraphy was 3.4 mm. All the high uptake samples and all but two of the intermediate uptake samples showed cancer, whereas histology found malignant tissue in only 2 out of the 8 low uptake samples. CONCLUSION: 99mTc BN confirms its high sensitivity in detecting breast cancer. IP is able to drive or co-drive breast biopsy when used with appropriate radiopharmaceuticals. PMID- 12894589 TI - ERF, an ETS-related transcriptional repressor, can induce erythroid differentiation. AB - Ets-family genes have been implicated in leukemia, as well as in normal hematopoiesis. ERF is an ets-related gene that represses transcription and is regulated by MAPK phosphorylation through its effect on ERF sub-cellular localization. Using pluripotent human cell lines, we studied the effect of ERF on erythroid differentiation. K562 and HEL cells expressing ERF expressed elevated levels of the erythroid-specific markers CD71 and Glycophorin A, as well as hemoglobin and GATA1. Treatment with the Erk kinase inhibitor PD98058 further enhanced the erythroid phenotype in ERF-expressing cells and cells expressing a non-phosphorylatable ERF mutant exhibited an even more enhanced phenotype. These results are consistent with the fact that ERF function is regulated by MAPK, and suggest that the effect of the MAPK pathway in erythroid differentiation is partially mediated by ERF. The effect of ERF is similar to the one shown for ETS1 and opposite to the FLI1 function in these cells, suggesting that several ets genes may play key roles in hematopoietic differentiation. PMID- 12894590 TI - Minireview: DEK and gene regulation, oncogenesis and AIDS. AB - A brief review summarizing the relationships that exist between the DEK protein and gene regulation, oncogenesis and AIDS. PMID- 12894591 TI - Friend virus-induced erythroleukemias: a unique and well-defined mouse model for the development of leukemia. AB - Retroviruses lacking oncogenes have been known to induce various types of cancer when inoculated into animals. Among these, Friend virus, discovered by Charlotte Friend in 1957, is capable of inducing erythroleukemias when injected into susceptible strains of mice. Since its discovery, this murine model of leukemogenesis has been extensively used to study the multistage nature of cancer. In the past two decades, several oncogenes and tumour suppressor genes, which play critical roles in the induction and progression of Friend erythroleukemia, have been identified. Retroviral insertional activation of Fli-1 and Spi-1/PU.1, as well as loss of tumour suppressor genes such as p53 or p45 NFE2 have been shown to be critical for the induction and progression of Friend virus-induced erythroleukemias. The majority of these genetic changes have also been implicated in various types of human neoplastic transformations. In this review we will discuss the genetic changes associated with Friend Disease, the temporal order during induction and progression of disease, and the function of these genes in both normal erythroid development as well as malignant transformation. PMID- 12894592 TI - Samarium-153 and technetium-99m-labeled monoclonal antibodies in angiogenesis for tumor visualization and inhibition. AB - BACKGROUND: Angiogenesis is the development of new blood vessels from pre existing ones. Vascular endothelial growth factor (VEGF) is one of the most important angiogenic activators. Our studies are focused on the detection of VEGF by use of radiolabeled anti-endothelial monoclonal antibodies, which have the ability to localize in newly-formed vasculature of a cancerous origin. MATERIALS AND METHODS: The anti-endothelial monoclonal antibody VG76e was labeled with Samarium-153 and Technetium-99m. Biodistribution of the radiolabeled species was assessed in normal female Swiss mice, while tumor uptake was also evaluated. RESULTS: VG76e was labeled with 99mTc and 153Sm, resulting in a single product with a labeling yield of over 95%. Biodistribution studies showed non-specific uptake in any organ, with elimination via the hepatobiliary system. Finally, satisfactory tumor uptake was observed for both radiolabeled derivatives. CONCLUSION: Monoclonal antibodies raised against epithelial growth factors or their receptors, when labeled with appropriate radionuclides, may be a useful tool for early tumor detection and eventually for therapy. PMID- 12894593 TI - Ets-2 interacts with co-repressor BS69 to repress target gene expression. AB - BACKGROUND: The ETS-family of proteins consists of over 30 members that regulate the growth, differentiation and survival of both normal and tumor cells. How specificity is achieved within this family remains largely unresolved. One mechanism for attaining specificity is through the action of signaling pathways on specific family members. For example, Ets-2 is an activator modulated by ras dependent phosphorylation of a single residue in the conserved pointed domain of this factor. We hypothesized that phosphorylation of the pointed domain regulates the proteins that can interact with ets-2 in the cell nucleus, resulting in regulation of target genes. MATERIALS AND METHODS: We used a combination of biochemical assays, yeast two-hybrid screens and transfection assays to identify and characterize proteins interacting with the pointed domain. RESULTS: BS69, a known co-repressor, was identified in a yeast two hybrid screen as an ets-2 interacting partner. BS69 can interact with ets-2 in vivo and phosphorylation of the ets-2 pointed domain decreased the interaction with BS69 in vitro. In transfection assays, co-expression of ets-2 and BS69 resulted in repression of defined ets-2 target genes. CONCLUSION: These results support a role for ets-2 as a repressor and indicate that BS69 is required as co-repressor. Phosphorylation of ets-2 may switch its activity from repressor to activator by interfering with formation of the BS69 complex. PMID- 12894594 TI - CRD-BP: a c-Myc mRNA stabilizing protein with an oncofetal pattern of expression. AB - The Coding Region Determinant-Binding Protein (CRD-BP) is an RRM and KH-domain containing protein that recognizes specifically at least three RNAs. It binds to one of the two c-myc mRNA instability elements, to the 5'Un Translated Region (UTR) of the leader 3 IGF-II mRNA and to the oncofetal H19 RNA. CRD-BP has been assigned a role in stabilizing c-myc mRNA by preventing its endonucleolytic cleavage and in repressing the translation of the leader 3 IGF-II mRNA, the major embryonic species of this message. CRD-BP is normally expressed only in fetal tissues. However, its expression is detected in primary tumors and transformed cell lines of different origins. The vast majority of colon (80%) and breast (60%) tumors and sarcomas (73%) express CRD-BP whereas in other tumor types, for example prostate carcinomas, its expression is rare. CRD-BP expression has also been detected in benign tumors such as breast fibroadenomas, meningiomas and other benign mesenchymal tumors, implying a role for this gene in abnormal cell proliferation. In breast carcinomas, CRD-BP expression and or gene copy number gains in the region encompassing the c-myc locus were detected in approximately 75% of tumors, implying that the deregulated expression of c-myc may be more widespread than previously believed. Infiltrated lymph nodes, corresponding to CRD-BP-positive primary tumors, were also found positive indicating that monitoring for CRD-BP could prove useful for the detection and monitoring of disseminated disease. PMID- 12894595 TI - Study of the mechanisms of cigarette smoke gas phase cytotoxicity. AB - BACKGROUND: The cytotoxicity of cigarette smoke (CS) in humans is well documented, but the mechanism behind CS toxicity and carcinogenicity remains unknown. We are interested in the toxicological effects of CS gas phase and the biological mechanisms of its action. MATERIALS AND METHODS: Gas phase CS cytotoxicity was measured by Wst-1 and LDH assays, in cultured cells. The mechanism of cell death was investigated by flow cytometric analysis using Annexin V and PI staining. Gas phase CS-induced oxidative damage was evaluated by estimating cellular glutathione (GSH) levels. Protein modifications (nitration of tyrosines) induced by gas phase CS and activation of key signalling proteins (Mitogen-activated protein kinase, MAPK) were detected by immunoblotting. RESULTS: The cytotoxicity of gas phase CS was found to be dose-dependent. The mechanism of cell death was found to be both apoptotic and necrotic depending on the concentrations used. Exposure to gas phase CS resulted in depletion of cellular GSH levels, increased nitrotyrosine immunoreactivity and phosphorylation of p44/42 MAPK proteins. CONCLUSION: These results suggest that the CS gas phase alone contributes significantly to the deleterious effects of CS in cellular systems. PMID- 12894596 TI - Genetic alterations of the APC gene in familial adenomatous polyposis patients of the hellenic group for the study of colorectal cancer. AB - Familial Adenomatous Polyposis (FAP)-a premalignant clinical entity inherited as an autosomal dominant trait-is characterized by the development of hundreds to thousands of adenomatous polyps of the colorectum during the second and third decade of life. Approximately 80% of the FAP patients harbour truncating germ line mutations in the APC tumor suppressor gene (Adenomatous Polyposis Coli). We tested 48 members from 9 families. Two novel truncating mutations were identified 2601delGA, R923X--and five already known mutations R564X, R876X, Q1045X, 3927 3931delAAAGA and D1822V were found. Our method for testing was PCR amplification from genomic DNA extracted from whole blood, followed by automated DNA sequencing. PMID- 12894597 TI - Biodistribution and scintigraphic studies of 153Sm-labeled anti-CEA monoclonal antibody for radioimmunoscintigraphy and radioimmunotherapy. AB - The anti-CEA monoclonal antibody, which selectively localizes in colon cancer, was labeled with Samarium-153 (153Sm). 153Sm is mainly a beta-emitter which can be used for therapeutic purposes, while its gamma-ray facilitates imaging studies. Labeling was achieved using the bicyclic anhydride of DTPA as chelator for Sm-153 tagging onto the antibody. [153Sm]anti-CEA was biologically evaluated in nude mice bearing tumors of different weight (0.5-2.5 g), at diverse time intervals (4-72 hours), by anatomic and imaging methods. Biodistribution studies showed slow blood clearance and high retention in the liver, kidneys and lungs. In nude mice bearing tumors of about the same weight, uptake increased with time, from 4 to 72 hours post injection (p.i.). Highest uptake was observed in 0.5-0.8 g tumors compared to those of 1.5-2.5 g. The results agreed with imaging studies performed on a gamma camera at 4 to 72 hours p.i. Tumor uptake depended on time and tumor weight. The tumor can be visualized 24 hours p.i. but, due to the high background, it can be clearly distinguished at 72 hours p.i. PMID- 12894598 TI - Support for bereaved owners of pets. AB - TOPIC: The bond that exists between people and their pets and its impact on physical and mental health. PURPOSE: To review the current literature and explore the clinical implications of bereavement related to pets. SOURCES: A comprehensive review of the bereavement, veterinarian, and agricultural literature related to attitudes and response patterns to pet and animal death. CONCLUSIONS: The death or loss of a beloved pet can be a life-changing event. PMID- 12894599 TI - Exploring boundaries in the nurse-client relationship: professional roles and responsibilities. AB - TOPIC: The creation and maintenance of boundaries as they pertain to the nurse client relationship. PURPOSE: To challenge readers to reflect on their fiduciary responsibilities by identifying boundary crossings and violations. SOURCES: Review of the literature and the authors' clinical and teaching experiences. CONCLUSIONS: Although self-awareness and monitoring, debriefing, and availing oneself of supervision and education are important tools in creating and maintaining boundaries, in the final analysis, the nursing profession needs nurses who have the ability to make decisions about boundaries based on the best interests of the clients in their care. PMID- 12894600 TI - Theory-based research in schizophrenia. AB - TOPIC: Theoretical models to guide research into schizophrenia. PURPOSE: To review the most commonly used biological and environmental models of schizophrenia. SOURCES: Published literature and research conducted by the author. CONCLUSIONS: The vulnerability model includes both biological and environmental variables that impact the course of schizophrenia and treatment. RESPONSES: Thus, it may be a useful framework for nursing research examining responses to various psychosocial treatments in schizophrenia. PMID- 12894601 TI - Three potentially fatal adverse effects of psychotropic medications. PMID- 12894602 TI - Acetylcholinesterase inhibitors in dementia. PMID- 12894604 TI - [SARS, possible zoonosis in the area of conflict of pathogenic coronaviruses of animals]. AB - Severe acute respiratory syndrome (SARS) is an emerging disease, which was first recognized in Guangdong Province, China, in November 2002. In the meantime, SARS has been recognized in patients on all five continents. A novel coronavirus, which is not related to the hitherto known coronaviruses, has been proven to be associated with the disease. Our genomic analyses strongly suggest that the new SARS-coronavirus did not emerge through mutation or recombination and that it has probably been transmitted from a so far not identified animal species to humans. Therefore, it is most likely that SARS virus is a zoonotic agent. A broad body of knowledge originating from research in veterinary medicine indicates that development of vaccines against the SARS-coronavirus may be problematic. The potential danger of such vaccines should not be neglected during the process of vaccine development. PMID- 12894605 TI - [Cortisol/creatinine ratio in urine (UCC) of healthy cats]. AB - In 31 healthy cats urine samples were taken to determine the cortisol/creatinine ratio (UCC) during hospitalisation and at home. The UCC of the samples, which had been taken in the clinic, was significantly higher (0-19 x 10(-6), Median 3 x 10( 6)) than the one of the samples taken at home (0-4 x 10(-6), Median 1 x 10(-6)). The parameter was neither influenced by the cat's age, sex or the fact that the cat stayed inside or outside, nor by the degree of visible agitation. Assay validation achieved good results regarding precision and accuracy of the measuring of cortisol and creatinine in the urine. The study shows that stress- caused by the visit to the veterinarian--can provoke a significant increase of UCC. Therefore, the parameter should be determined only from urine samples taken at home. Furthermore, it is important to notice that cortisol metabolites are measured in varying degree with the different assays. Therefore, it is inevitable that each laboratory generates its own reference values. PMID- 12894606 TI - [Suckling mortality in fattening rabbits in Switzerland]. AB - The aims of this project were to determine mortality rate and aetiology of losses in suckling rabbits during the first 14 days of life. Post-mortem examinations were done on 1096 siblings from 8 conventional farms. Mortality rates varied from 0 to 17.0%. Main death causes were non-infectious and included stillbirth (13.9%), weakness due to reduced birth weight (3.0%), starvation (11.6%), runting (11.6%), cannibalism (0.5%) and scattering (7.9%). The last four causes mainly attributed to insufficient nursing by the doe. Only 21.8% of the losses were of infectious aetiology. PMID- 12894607 TI - [Melanosis maculosa in a calf]. AB - This work describes findings in a fattened calf, which were seen in a routine slaughter. The most important findings by the meat inspector were diffuse, black spots between 2 and 70 mm in diameter. This hyperpigmentation was observed on the entire carcass as well as on the outside and inside of all internal organs. Removal of the spinal cord produced black colored liquor. Subsequent microbiological examination of the meat resulted in sterile findings. The muscle pH was 5.8. On histological examination many macrophages containing melanin were found in the lung and liver. Based on these findings the diagnosis of Melanosis maculosa was made. PMID- 12894608 TI - [Sonographic detection of an echogenic thrombus in the vena cava caudalis in a cow]. PMID- 12894609 TI - Panic in the emergency room. AB - OBJECTIVE: This paper examines the relation between coronary artery disease (CAD) and panic disorder (PD), discusses the implications of this relation to the general medical system, and suggests future assessment and intervention strategies for emergency departments. METHOD: We reviewed the literature on CAD and PD using Medline and PsycINFO. RESULTS: PD is more expensive to our nonpsychiatric, general medical system than any other psychiatric condition. The main reason for PD patients' continued use of general medicine for their psychological symptoms is that their PD remains undiagnosed. In the emergency room (ER), PD patients with chest pain have their PD go undiagnosed about 98% of the time. By having ERs implement specific assessment and intervention strategies for patients presenting with chest pain, the savings to the general medical system could be substantial. CONCLUSIONS: By improving recognition of PD in the ER, there is the potential to generate large savings in general medical care. With the availability of empirically supported or effective psychological and pharmacologic treatments for PD, appropriately diagnosing and subsequently treating patients with PD may prevent them from experiencing many years of disability and higher rates of fatal coronary events. PMID- 12894610 TI - Medication strategies in childhood aggression: a review. AB - OBJECTIVE: To review studies of psychopharmacological management of aggression in common childhood psychiatric disorders. METHOD: Using OVID software, we searched Medline for studies that were undertaken in the last 30 years. Controlled and uncontrolled data are summarized for each condition. RESULTS: A paucity of evidence-based information currently exists. Even so, specific indications from the existing literature can be suggested for several classes of psychotropics, particularly in conduct disorder (CD), attention-deficit hyperactivity disorder (ADHD), mood disorders, and other conditions. CONCLUSIONS: Clinicians can use findings from reviewed controlled and, where necessary, uncontrolled studies to inform pharmacologic practice. This review offers suggestions for future research directions that will aid clinical practice. PMID- 12894611 TI - Old and homeless: a review and survey of older adults who use shelters in an urban setting. AB - OBJECTIVES: Research on the mental health and service needs of homeless seniors has been scant. This paper reviews the available literature and presents findings of a Toronto survey in an effort to describe the demographics of homeless seniors, their level of impairment, and their mental and physical health needs. METHODS: We searched the Medline, AgeLine, and PsycINFO databases, using the following key words: elderly homeless, elderly hostel users, and urban geriatrics. To better describe the service needs of the elderly homeless, we obtained demographic data from the Community and Neighbourhood Services Department and distributed a survey questionnaire to 11 Toronto hostel directors. The questionnaire elicited data relating to reasons for shelter use, problem behaviours, and mental health needs of those over age 65 years. RESULTS: Although seniors represent a small percentage of the homeless population, their numbers are growing. The available literature suggests a high prevalence of psychiatric disorders and cognitive impairment in this population, with a greater proportion of older women than men having severe mental illness. Further, our survey suggests that the service needs of elderly hostel users in Toronto differ from those of their younger counterparts. CONCLUSION: The homeless elderly are the most vulnerable of this impoverished population. Although more research is needed to define their mental and physical health needs and ways of meeting them, their characteristics appear to be unique. Geriatric psychiatrists could play a significant role in evaluating and treating this population more comprehensively. PMID- 12894612 TI - Correlates of therapeutic response in panic disorder presenting with palpitations: heart rate variability, sleep, and placebo effect. AB - OBJECTIVE: To examine the correlates of therapeutic response of patients with panic disorder presenting with palpitations, we hypothesized that therapeutic response would correlate with heart rate variability (HRV) and sleep measures. METHODS: After a 1-week placebo washout, 27 patients free of structural heart disease and not on cardioactive drugs were randomized in a double-blinded fashion to 4 weeks of treatment with clonazepam (a known antipanic agent) or placebo. We performed standard sleep measures and recorded HRV from 24-hour Holter acquisitions at baseline and end of study. We defined response to therapy as a 50% improvement in the Hamilton Anxiety Rating Scale (HARS) score, confirmed by questionnaires and reaction to sodium lactate infusion. RESULTS: There were 12 responders and 15 nonresponders. Normalization of sleep pattern (including less stage 1 and rapid eye movement [REM] sleep) was observed in both drug and placebo responders (P = 0.011 and P = 0.05, respectively) and in placebo responders alone, compared with nonresponders (P = 0.006 and P = 0.013, respectively). Placebo responders were more likely to show less depression, but even after we controlled for depression, main sleep effects remained. None of the HRV measures correlated with response, but compared with placebo, clonazepam led to a decrease in all the time and frequency domain measures of HRV (all P < 0.05). CONCLUSIONS: Central mechanisms are related to the therapeutic response of patients with panic disorder presenting with palpitations, but this does not directly correlate with HRV. Larger and longer studies may allow objective explanations of placebo response in panic disorder. PMID- 12894613 TI - Randomization in a substance abuse treatment study: participants who consent vs those who do not. AB - OBJECTIVE: A mixed research design study compared patients who consented to treatment randomization with patients who opted for usual care. METHODS: Patients in substance abuse rehabilitation who consented to randomization (that is, consenting group [CG]) were compared with those unwilling to undergo randomization (that is, nonconsenting group [NG]) but who, nevertheless, underwent the experimental assessment sessions, which spanned from intake to 6 month follow-up. RESULTS: Patients in the CG exhibited a longer history of drug abuse, less occupational stability at intake, and following intensive treatment, tended not to do as well in terms of recovery, compared with those in the NG. CONCLUSION: Inclusion of data from partial participants in research protocols may offer a viable strategy to better appraise data generalizability. PMID- 12894614 TI - Fright (effroi) and other peritraumatic responses after a serious motor vehicle accident: prospective influence on acute PTSD development. AB - OBJECTIVE: We prospectively examined the relation between various peritraumatic responses (that is, fear, helplessness, horror, amnesia, and fright) and the development of posttraumatic stress disorder (PTSD) 2 months after a trauma. METHOD: Participants included 123 motor vehicle accident (MVA) victims consecutively hospitalized in a traumatology department for over 72 hours during a 16-month period. Between day 2 and day 5 of their hospitalization, a psychiatrist assessed the patients' peritraumatic responses and acute stress disorder (ASD). Two months after the accident, an experienced psychiatrist contacted the patients by telephone, and the PTSD symptoms and the diagnosis were assessed with a modified version of the Clinician-Administered PTSD Scale (CAPS). RESULTS: Of the participants, 48 reported an immediate fright reaction when faced with the prospect of their own death. Participants who reported a fright experience had a 17 times greater risk of subsequently meeting the diagnostic criteria for PTSD (odds ratio 16.75). A fright reaction predicted PTSD development with a specificity of 0.93 and a sensitivity of 0.60 CONCLUSION: The immediate reactions described in criterion A2 of the DSM-IV (that is, fear, helplessness, and horror) did not seem to be equally relevant. An initial feeling of fright seems to be an essential qualitative factor in the clinical description of psychological trauma. PMID- 12894615 TI - Children with psychiatric disorders: the A1 Ain Community Psychiatric Survey. AB - OBJECTIVE: The prevalence of child psychiatric morbidity in the community is unknown in most developing countries, including those in the Arab region. METHOD: An epidemiologic study was carried out to estimate the prevalence of psychiatric morbidity and to determine the sociodemographic correlates in a sample of children in the community, aged 6 to 18 years, in A1 Ain, United Arab Emirates (UAE). RESULTS: We obtained a prevalence rate of 22.2% for overall morbidity, as classified in the DSM-1V, and 14.3% for those with significant dysfunction, with the most common diagnosis being mood disorders. Female sex, large family size, chronic life difficulties, family history of psychiatric disorder, and alcohol related problems in a family member were significantly associated with DSM-IV diagnosis. CONCLUSION: Although the prevalence and symptomatology in this Middle East community are similar to those in Western studies, none of these children had received professional help, suggesting serious deficiencies in mental health care services in the country. PMID- 12894616 TI - A follow-up study of persons found not criminally responsible on account of mental disorder in British Columbia. AB - OBJECTIVE: To describe the characteristics of individuals found not criminally responsible on account of mental disorder (NCRMD) after the 1992 Criminal Code amendments and to track their movement through the forensic system, as well as to unveil the changes to British Columbia's forensic psychiatric system that resulted from Bill C-30. METHOD: Profile information relating to persons found NCRMD between February 4, 1992, and February 4, 1998, in British Columbia was collected and analyzed. Community follow-up data was collected and analyzed for a 24-month period following a subject's discharge from hospital. RESULTS: A substantially greater number of individuals entered the forensic psychiatric system in British Columbia after Bill C-30 was implemented. The post-1992 forensic psychiatric population contained fewer persons charged with serious index offences and a greater number of persons charged with relatively minor offences. The length of hospitalization for the NCRMD cohort decreased substantially after the 1992 Criminal Code amendments. CONCLUSION: The Bill C-30 provisions have made the NCRMD defence an attractive option for defendants and legal counsel. PMID- 12894617 TI - Anxiety-related visits to Ontario physicians following September 11, 2001. AB - OBJECTIVE: To determine whether the climate of increased anxiety following the terrorist attacks of September 11, 2001, and the subsequent anthrax cases led to increased anxiety-related physician visits. METHOD: We undertook a retrospective, population-based study of all Ontario residents. We identified physician visits using Ontario's universal health care insurance program. Interventional autoregressive integrated moving average (ARIMA) time series models were used to examine the impact of the terrorist attacks and anthrax infections on the rate of anxiety-related physician visits. RESULTS: Neither the terrorist attacks of September 11 (P > or = 0.40) nor the anthrax infections in October and November (P > or = 0.14) had a statistically significant impact on the rate of anxiety related visits to physicians. CONCLUSIONS: The climate of heightened anxiety did not result in increased demand for mental health services. PMID- 12894618 TI - Risperidone liquid treatment of acute mania. PMID- 12894619 TI - Methylphenidate and the cytochrome P450 system. PMID- 12894620 TI - Antiaggressive action of combined risperidone and quetiapine in a patient with schizophrenia. PMID- 12894621 TI - Ultrarapid response to an antidepressant: a clue to bipolarity? PMID- 12894622 TI - Developmental alcohol exposure, circadian rhythms, and mood disorders. PMID- 12894623 TI - [Sexual touching practiced by an adolescent substituted by testosterone]. PMID- 12894624 TI - Dichotomization and manipulation of numbers. PMID- 12894625 TI - Maslow's Hierarchy of Needs--revisited. PMID- 12894626 TI - A concept analysis of susceptibility: application of the hybrid model. AB - TOPIC: Choosing to obtain a screening mammogram is often determined by a woman's perceived susceptibility. PURPOSE: To establish the importance of susceptibility to breast cancer in rural women in southeast Louisiana, using concept analysis as the hybrid model. SOURCES: Through this analysis, findings from the literature were compared to women's actual perceptions from the field. CONCLUSIONS: The author concludes that rural women in Louisiana do not view breast cancer as a potential harm, nor is there a belief that they have control over the decision to have a screening mammogram; therefore, rural women do not perceive themselves susceptible. PMID- 12894627 TI - Preserving dignity in the young hospitalized child. AB - TOPIC: Bodily intrusion and exposure are common experiences for hospitalized children. Placing children in such compromising situations can have a significant impact on their self-perception. PURPOSE: To recognize pediatric patients' inherent powerlessness in the hospital setting and suggest strategies to overcome it. SOURCES: Published literature and author's clinical experiences. CONCLUSIONS: Nurses have a key role in ensuring that the psychosocial issues of privacy, modesty, and self-esteem are recognized, respected, and incorporated appropriately into their clinical practice when caring for hospitalized children. PMID- 12894628 TI - To make a difference: nursing presence. AB - TOPIC: Nursing presence in clinical practice. PURPOSE: To explore the concept of presence within the nursing literature. SOURCES: Published literature. CONCLUSIONS: True presence creates the opportunity for nurses to go where the patient is in life, to learn about the experience of health as it is defined and lived, and to work with patients as they choose the meaning of the situation. PMID- 12894629 TI - Palliative care: a discussion of management and ethical issues. PMID- 12894630 TI - Life and death in tribal Africa. PMID- 12894631 TI - Significance of functional status data for payment and quality. AB - To date, the Medicare Program has used functional status information (FSI) in patient assessment tools, performance assessment, payment mechanisms, and--most recently--in quality measures to inform consumer choice. This article explores the rationale for the collection of functional status data to promote innovative models of care and examines issues related to data collection for quality improvement, performance measurement, and payment. In this issue of the Health Care Financing Review, articles focus on collection and classification of functional status for payment and quality purposes. PMID- 12894632 TI - Comparison of functional status tools used in post-acute care. AB - There is a growing health policy mandate for comprehensive monitoring of functional outcomes across post-acute care (PAC) settings. This article presents an empirical comparison of four functional outcome instruments used in PAC with respect to their content, breadth of coverage, and measurement precision. Results illustrate limitations in the range of content, breadth of coverage, and measurement precision in each outcome instrument. None appears well-equipped to meet the challenge of monitoring quality and functional outcomes across settings where PAC is provided. Limitations in existing assessment methodology has stimulated the development of more comprehensive outcome assessment systems specifically for monitoring the quality of services provided to PAC patients. PMID- 12894633 TI - Measuring function for Medicare inpatient rehabilitation payment. AB - We studied 186,766 Medicare discharges to the community in 1999 from 694 inpatient rehabilitation facilities (IRF). Statistical models were used to examine the relationship of functional items and scales to accounting cost within impairment categories. For most items, more independence leads to lower costs. However, two items are not associated with cost in the expected way. The probable causes of these anomalies are discussed along with implications for payment policy. We present the rules used to construct administratively simple, homogeneous, resource use groups that provide reasonable incentives for access and quality care and that determine payments under the new IRF prospective payment system (PPS). PMID- 12894634 TI - An assessment tool translation study. AB - Policymakers hoped to substitute a new, multi-purpose, functional assessment instrument, the minimum data set post-acute care (MDS-PAC), into the planned prospective payment system (PPS) for inpatient rehabilitation hospitals. PPS design requires a large database linking treatment costs with measures of the need for care, so the PPS was designed using the functional independence measure (FIM) database linked to Medicare hospital claims. An accurate translation from the MDS-PAC items to FIM--like items was needed to ensure payment equity under the substitution. This article describes the translation efforts and some of the problems that led policymakers to abandon the effort. PMID- 12894635 TI - Capturing and classifying functional status information in administrative databases. AB - The health care delivery system aims to improve the functioning of Americans, but little information exists to judge progress toward meeting this goal. Administrative data generated through running and overseeing health care delivery offer considerable information about diagnoses and procedures in coded formats comparable across settings of care. This article explores the issues raised when considering adding coded information about functional status to administrative databases throughout the health care system. The National Committee on Vital and Health Statistics (NCVHS) identified the International Classification of Functioning, Disability and Health (ICF) as the only viable code set for consistently reporting functional status. PMID- 12894636 TI - WHO's ICF and functional status information in health records. AB - A common framework for describing functional status information (FSI) in health records is needed in order to make this information comparable and of value. The World Health Organization's (WHO's) International Classification of Functioning, Disability and Health (ICF), which has been approved by all its member States, provides this common language and framework. The biopsychosocial model of functioning and disability embodied in the ICF goes beyond disease and conceptualizes functioning from the individual's body, person, and lived experience vantage points, thereby allowing for planning interventions targeted at the individual's body, the individual as a whole or toward the environment. This framework then permits the evaluation of both the effectiveness and cost effectiveness of these different interventions in devising programs at the personal or societal level. PMID- 12894637 TI - Functional status and health information in Canada: proposals and prospects. AB - The primary obstacle to evidence-based health care quality assessment in Canada is reliable data on health encounters and episodes of care. The recent Federal/Provincial Health Accord will enhance health data collection, including standardized functional status information (FSI) for administrative records. Canadian health policy developers also agree that FSI is needed to bridge data gaps since alterations in functional status create the continuity that links all episodes of care and health service utilization. Given Canada's universal, single payer, health financing structure, the prospects for coherent and systemwide data collection are good. This article describes the Canadian health care from the perspective of health information, and surveys proposals in electronic health technology development, the obstacles that need to be faced, and the prospects of doing so. PMID- 12894638 TI - From clinical records to regulatory reporting: formal terminologies as foundation. AB - Achieving goals of healthy people and populations is dependent on available and relevant data for health care decisions. New technologies enable reuse of data for decisions, however it is clear that uniform data standards and in particular standards around terminological data will be required to achieve reuse. Terminological data related to functioning and disability presents unique challenges because of the conceptual ambiguity within the field. The International Classification of Functioning, Disability, and Health (ICF) provides a clarifying conceptual foundation for functioning and disability data, but is not structured as a formal terminology. The need for a concerted and coordinated effort is emphasized. PMID- 12894639 TI - Medigap reform legislation of 1990: a 10-year review. AB - The 1990 Medigap reform legislation sought to make it easier for consumers to compare policies, provide market stability, promote competition, and avoid adverse selection. Evidence is that the standardization of benefits has simplified consumer choice and is strongly supported by consumers and State regulators. The 1990 reforms also decreased carrier and agent abuses. However, loss ratios (the proportion of premiums paid in benefits versus being retained for administration and profit) have changed little since 1990, bringing into question whether price competition has been enhanced. The prescription drug benefit, which is included in 3 of the 10 standardized plans, provides only limited financial protection yet is expensive, one reason being adverse selection. Access to coverage for Medicare disabled beneficiaries is problematic in most States. PMID- 12894640 TI - Drug coverage, utilization, and spending by Medicare beneficiaries with heart disease. AB - For Medicare beneficiaries who report having heart disease, drug coverage and type of supplemental health insurance affect the likelihood of usage and costs of heart medications, but not the extent of usage. Nearly one in five does not use heart medications and of the latter, one-third lack drug coverage. Some non-users without drug coverage go without prescribed heart medications because of limited financial access. Compared to non-users with coverage, they utilize medical provider services more, and if hospitalized, their inpatient costs are twice as high. Medicare may accrue cost savings by providing drug coverage to and monitoring these at-risk beneficiaries. PMID- 12894641 TI - Method of friction coefficients determining for ternary solutions of non electrolytes and polymeric membrane. AB - In this paper, the dependencies between phenomenological coefficients for membrane and friction coefficients for solutes and membrane in the framework of generalised Spiegler-Kedem-Katchalsky model for non-ionic and homogeneous solutions were presented. The matrixes of transformation from phenomenological onto frictional coefficients and from frictional onto phenomenological coefficients were worked out. Besides, the influence of hydration number of solutes on friction and phenomenological coefficients was elaborated. Moreover, the friction coefficients and their dependencies on hydration number of glucose for aqueous glucose solution (binary solution) and glucose solution in aqueous ethanol solution (ternary solution), permeating through flat polymer membrane were presented. PMID- 12894642 TI - [Comparative evaluation of TNF-alpha induction in vitro and in local tissue reaction after implantation of HAP/TCP and HAP]. AB - The aim of this work was the evaluation of biocompatibility and bioactivity of HAP/TCP as compared to HAP. This evaluation was carried out by the in vitro assessment of the induction of TNF-alpha in the rat blood after the contact with the tested HAP. The anther target was to investigate of local muscle tissue reaction of rats bones reaction of rabbits, after the implantation of tested materials. The level of TNF-alpha was estimated by biological method in the supernatants of rat blood cell cultures stimulated in vitro by tested materials. The implantation assessment was carried out on 25 rats and 15 rabbits. The muscle tissue reaction was evaluated after 2, 4, 8, 12, and 24 weeks, while the reaction of bones after 4, 12, and 26 weeks since the implantation. The level of TNF-alpha in the blood after stimulation in vitro with HAP/TCP was on the same level as control group. The implantation of HAP the level of TNF-alpha was twice as high as compared to HAP/TCP and control groups. These differences were of statically importance. The muscle tissue reaction after implantation of HAP/TCP was characterized by moderate inflammation during the early period which lead to the formation of two layer connective tissue capsule. In the capsule wall there were great cells of foreign body type and macrophages. In the cytoplasm of some the cells particles of HAP were noted, what could be regarded as the proof of its resorption. In the bone tissue surrounded HAP/TCP implants the stronger osteoconductive proprieties were observed as compared to the tissue around HAP. PMID- 12894643 TI - Tissue reaction after implantation of Spongostan, as the carrier of xenogenic bone morphogenetic protein. Experimental study. AB - Reconstruction or filling of bone defects, especially in the maxillofacial region, often requires use of biomaterials. An implant should fasten healing of the bone gap or it should replace autogenic bone grafts. The combination of bone morphogenetic proteins with suitable carrier may fulfill these requirements. Proteins causing differentiation of mesenchymal cells in chondroblasts and osteoblasts were called Bone Morphogenetic Proteins--BMPs. The authors extracted BMP from bovine bones and placed it into collagen carrier formed from generally accessible hemostatic sponge--Spongostan. The implants were grafted into rat femoral muscle pouches in order to trace the tissue response. Pathologic examinations were performed 3, 6 and 8 weeks after implantation. On the basis on macroscopic and microscopic examinations it was stated that collagen sponge speckled with BMP caused minimal tissue response and evolved characteristic thin connective tissue capsule formation around the implant. The connective tissue penetrated spongious structure of the implant, filling the spaces, which became growing due to sponge resorption. Characteristic hyalinization and sparse chondroblasts were visible 8 weeks after implantation. PMID- 12894644 TI - Own observations and experiences on usage of artificial prosthesis--Dallon H in vascular operations. AB - Author on the ground 124 of artificial grafts Dallon H grafted in years 2000- June 2002 in Department of Vascular, General and Transplantation Surgery in Wroclaw with protracted ischaemia of lower limbs, represents own experiences for observations of useful features this artificial graft. The hydrophility of these grafts, at maintenance all of other comparable features with grafts of westerly firms causes, that Dallon H I gained oneself large acknowledgement as many-sided artificial grafts alternative to vascular prosthesis of renowned westerly firms. PMID- 12894645 TI - The treatment of massive prosthetic grafts infections with the use of silver/collagen coated dacron vascular prosthesis. AB - In this paper the use of silver-coated medical prosthesis of massive aorto bifemoral and ilio-femoral dacron graft infection with hemorrhage is presented. To solve this problem in situ replacement was attempted using silver-coated dacron vascular prosthesis more resistant to infection. 9 patients with this therapeutic method is reported. Although positive clinical effect was obtained, the scintigraphy and Duplex Doppler Ultrasound of implanted graft revealed its infection. In our opinion therefore application of silver-coated medical prosthesis should be very cautious and limited to the critical states. PMID- 12894646 TI - [Use of chitosan in medicine and biomedical engineering]. AB - Applicability of different forms of chitosan in medicine and biomedical engineering has been discussed on the basis of a literature survey. Attention was paid to the use of chitosan as a drug carrier and as a pharmacological agent in the treatment of hypercholesterolemia. Suitability of the polymer was proved in the process of blood purification (LDL-apheresis, haemoperfusion, dialysis) and wound healing (dressings). A possibility of using it in biohybrid artificial organs and as athrombogenic surfaces was also mentioned. PMID- 12894647 TI - A community health assessment of the North Mississippi Health Services Catchment Area. PMID- 12894649 TI - SARS, biological terrorism, and mother nature. PMID- 12894650 TI - Looking back: allergy in 1917: words of a "crank on food intoxication". PMID- 12894651 TI - Information and Quality Healthcare--expansion of Medicare Beneficiary Protection. PMID- 12894652 TI - A perioperative work environment based on values. Can it be so? PMID- 12894653 TI - Oh, those baby blues: when you are dealing with a child with PHPV. PMID- 12894654 TI - Why a RN first assistant? A look at the benefits.... PMID- 12894655 TI - Bedside cystometry--a useful diagnostic tool for nurse continence advisors. PMID- 12894656 TI - Nursing education and gerontological nursing: fostering art and science or causing stigmas to fester? PMID- 12894657 TI - Issues of intimacy in dementia care. PMID- 12894658 TI - Promoting oral hygiene: a pilot project partnership. PMID- 12894659 TI - Making a difference: nursing students' perspectives on gerontological nursing. PMID- 12894660 TI - [Chlorhexidine varnishes in dentistry]. AB - Since the discovery of chlorhexidine as a potential surface antiseptic in the fifties, it has been abundantly used in different kinds of dental pharmaceutics. During the past decade chlorhexidine containing varnishes have been added to this list. These products present several advantages in comparison to the existing chlorhexidine formulations. Varnishes should be considered as further options for prevention of fissure and approximal caries, possibly in combination with already established methods. For patients at high risk for root surface caries, varnish application might be a meaningful preventive and therapeutic approach. In this paper, the existing literature on the use of two different chlorhexidine varnishes (Cervitec and EC40) in different fields in dentistry will be reviewed. PMID- 12894661 TI - [Materials for mouth protectors]. AB - Taking into account the number of teeth which are yearly irreversible traumatised during sport activities, the general use of mouthguards would contribute positively to the prevention of dental injuries. Custom-made mouthguards are more comfortable to wear and offer better retention and protection than stock and mouth-formed mouthguards. Different kinds of materials are available on the market for the construction of mouthguards. A polyethylene-polyvinylacetate copolymer (EVA) is the most suitable material. EVA allows the inclusion of hard or soft layers within the mouthguard. The thickness of a mouthguard is important for the reduction of applied forces to teeth: energy absorption capacity increases with material thickness. Increased thickness however, is associated with a reduction of comfort. Therefore, it is important that dentists take the patients' wishes and demands on both comfort and protection into consideration. A description of the clinical and technical method for the construction of a custom made mouthguard is given. PMID- 12894662 TI - [Calcium hydroxide-based pastes used in endodontics]. AB - The aim of the present study is to propose a classification of calcium hydroxide pastes on the basis of the vehicle that retains the calcium hydroxide. The vehicle determines the working characteristics of the calcium hydroxide: viscous and only vehicles prolong the action of the calcium hydroxide as compared with water-soluble substances. Physicochemical properties of the vehicle may limit the effectiveness in disinfecting the entire root canal system. As calcium hydroxide is not effective against all bacterial species found in root canal infections, associations with other medicaments have been proposed to enhance the efficacy of the intracanal medication. PMID- 12894663 TI - [Accidental extrusion of calcium hydroxide-based pastes into periapical lesions]. AB - Calcium hydroxide is a widely used material in endodontic treatment due to its high alkalinity and antimicrobial properties. It is thought to create a favourable environment for periradicular repair and healing, and to stimulate hard tissue formation. The aim of the present retrospective study was to evaluate the effect of accidental and voluminous overextensions of calcium hydroxide pastes into periapical lesions and tissues on the prognosis of periapical healing. It was seen that calcium hydroxide overextensions did not disfavour healing, none of the endodontic treatments failed but healing was delayed in the majority of cases and in a number of cases extrusion induced immediate flare-ups. Therefore the deliberate extrusion of calcium hydroxide into periapical tissues is not advocated. PMID- 12894664 TI - Facility profile. Facility touts advanced ORs and patient comfort. The Denton A. Cooley building, Houston. PMID- 12894665 TI - Touching the future. New technologies bolster existing security systems. PMID- 12894666 TI - Head of the class. ASHE's certificate program provides knowledge on the fine points of health care construction. PMID- 12894667 TI - Illuminating thoughts. Devising lighting strategies for clinical spaces. PMID- 12894668 TI - Safety first. New hospital design to put premium on protecting patients. PMID- 12894669 TI - Wide angle view. NFPA World Safety Conference covers a variety of issues. PMID- 12894670 TI - All accounted for--handling the daily fluctuations of today's hospital census. PMID- 12894671 TI - [Heparin-induced thrombocytopenia: 2002]. PMID- 12894672 TI - [Non-pharmacologic treatment of atrial fibrillation]. AB - The authors provide an update on non-pharmacological treatment of atrial fibrillation (AF). They emphasize that although antiarrhythmic drugs continue to be first-line therapy for the arrhythmia considered to be a cardiovascular epidemic, clinical research to develop non-pharmacological means of treatment has been unprecedentally intensified during the last decade. Electrical cardioversion is the most successful non-pharmacological method to restore sinus rhythm, also the efficacy and safety of AV node ablation for palliative ventricular rate controll is established. "Hybrid" therapeutic procedures, involving combinations of pharmacological and non-pharmacological interventions have gained widespread use. Curative transcatheter ablation for arrhythmia prevention is to be considered in case of clinical suggestions that AF is initiated by a primary regular arrhythmia that is amenable to routine catheter ablation (secondary AF). Despite encouraging results, at this point in time, curative catheter ablation for primary AF may offer significant improvement or even cure only for a small subset of patients, mostly young individuals with normal heart, and paroxysmal AF with frequent, symptomatic episodes refractory to multiple antiarrhythmic drugs. These interventions are to be performed in the settings of a clinical research project in some institutions. Regarding pacemaker therapy in case of bradycardia indication, physiologic pacing (AAI or DDD) is associated with significantly lower incidence of atrial fibrillation than ventricular pacing. Large-scale randomized controlled trials are needed to assess the clinical value of specially designed implantable devices to prevent atrial fibrillation in patients with no conventional bradycardia indication. Also, technical optimization and proper clinical evaluation is needed for implantable atrioverters and implantable cardioverter defibrillators capable of atrial cardioversion therapy. PMID- 12894673 TI - [Micro and mini-cholecystectomies in the 21st century]. AB - INTRODUCTION: Today the removal of the gallbladder is the safest, the most effective and widely recommended treatment for gallstone disease. Three essential methods are used for the removal of the gallbladder: standard open cholecystectomy, laparoscopic cholecystectomy and minicholecystectomy. Traditionally, the surgical community has resisted accepting minicholecystectomy. AIM: It is the Author's objective to illustrate the advantages and disadvantages of laparoscopic and microlaparotomy cholecystectomy. METHODS: The 2400 unselected patients who were operated with micro- and minicholecystectomy by them and a review of the relevant data of laparoscopic and standard minicholecystectomies permit some inferences to be made about the validity of microlaparotomy cholecystectomy. CONCLUSIONS: Main outcome measures regarding to mortality (0.12%), common bile duct injuries (0.08%), conversion an incision longer than 8 cm (0.29%) and syncronical choledocholithotomy (5.5%) as well as complete cholecystectomy (98.1%) indicate that micro- and minilaparotomy cholecystectomy with suitable technique and equipment are safe, less expensive choice either than the laparoscopic or the standard open cholecystectomy. PMID- 12894674 TI - [Failure of Helicobacter pylori eradication--suggestions for further therapy]. AB - Success of first H. pylori eradication attempts in the literature is around 80 90% and based on urea breath test of 1027 patients in Hungary is 75%. Repeated eradication attempts are needed in 10-25% of cases. In the clinical practice in Hungary second and third eradication attempts were successful only in 36% and 20% of cases. To improve efficacy the following suggestions has to be kept in mind: 1. Do not repeat the same combination if the first attempt is failed. 2. After failure of the first PPI + amoxicillin + clarithromycin triple therapy, either the quadriple therapy (PPI + tetracycline + metronidazole + bismuth) or the replacement of PPI with ranitidine bismuth citrate in the triple therapy is suggested. 3. If PPI + amoxicillin + metronidazole/tinidazole therapy fails, the metronidazole/tinidazole can be replaced by clarythromycin. 4. Do not start with clarithromycin + metronidazole/tinidazole therapy. 5. In case of uncertain previous therapies and for third eradication treatment send the patient to specialist. Rifabutin-based combinations seem to be effective, but the use of them in general practice is not advised due to the possible development of mycobacterium tuberculosis resistance. PMID- 12894675 TI - [Treatment of alcoholic liver disease]. PMID- 12894676 TI - [Hungarian patron of the Polish Academy of Sciences]. PMID- 12894677 TI - [Urban waste management--100 years ago]. PMID- 12894678 TI - [Zinc metabolism--a factor in canine aggression?]. AB - In order to test the hypothesis of zinc-deficiency as a factor in canine aggression, we examined sera of dangerously aggressive dogs and of behaviourally normal (non-aggressive) dogs for their zinc-contents. The results showed distinctly higher zinc-concentrations (mean +/- SD) in aggressive dogs (1.69 +/- 0.49 micrograms/ml) than in normal non aggressive dogs (0.76 +/- 0.16 microgram/ml). PMID- 12894679 TI - [Diagnosis of botulism since 1995. Report of test results]. AB - Since 1970 our laboratory is specialized in diagnosis of clostridial diseases, including Clostridium botulinum and botulism. Since 1995, samples from more than 900 suspected botulinal cases were received, mainly in cattle, horses and men. 524 outbreaks were diagnosed as clearly positive by toxin neutralisation; 83 cases remained inconclusive with the toxin neutralisation. The geographical distribution of the positive cases in Germany is demonstrated for cattle and horses. Dispatch and treatment of specimens and interpretation of results are discussed. PMID- 12894680 TI - [Distribution of immunoglobulin isotypes and Salmonella antibodies in blood serum and meat juice of pigs]. AB - Using the close linear regression between the logarithm of the dilution degree of a sample and the logarithm of the extinction measured in an ELISA both the relative concentrations of immunoglobulines of the isotypes IgG, IgM and IgA and of the LPS antibodies against S. Typhimurium of the different isotypes in blood sera and meat juice of 15 slaughtered pigs were detected and compared. Furthermore the total concentration of antibodies against LPS of S. Typhimurium according to the "meat juice ELISA" were compared. Distribution of immunoglobulines between serum and meat juice revealed individual differences between the animals as well as between the different immunoglobulin-isotypes. Within the same isotype the ratio of the concentrations of anti-LPS Salmonella Typhimurium antibodies between serum and meat juice was significantly closer than relating the whole of immunoglobulines of the referred isotype. In order to detect pig herds with a high level of Salmonella exposure a comparison of the 1:30 diluted meat juice samples with the 1:400 diluted blood sera is justified, however, for detailed epidemiological or scientific studies there is a need to consider the existing differences between the immunoglobuline-isotypes as well as between the specificity of antibodies and of total immunoglobulines. While the concentration of Salmonella antibodies of the isotypes IgG1, IgG2 and IgA showed a clear and statistically significant correlation between both one below the other and with the total amount of Salmonella antibodies, this connection could not be established for the total amount of immunoglobulines of different isotypes and the IgM-antibodies. PMID- 12894681 TI - [Usefulness of serological examinations in the analysis of Salmonella infections in pig herds]. AB - The development of the antibody concentration against lipopolysaccharide (LPS) of S. Typhimurium und S. Choleraesuis in rearing pigs during the fattening period and in breeding sows of the corresponding age was recorded. The studies revealed the following results. Antibodies of isotypes IgG1 and IgG2 revealed a more pronounced specificity against the according Salmonella serovar than IgM antibodies. The calculated "antibody percent value" based on the total amount of Salmonella antibodies is mainly determined by the IgM antibodies in sera and meat juice, respectively. In fattening pigs a significant increase of antibodies against IgM and total Ig was observed between week 3 and 10 after beginning of the rearing period. In breeding pigs this increase was detectable already earlier. In only 3 out of 10 groups an increase of IgG1 and IgG2 was also seen. The detected significant increase of total Ig and IgM in the other groups might be the result of a less intensive exposure to salmonellas or it might be due to an increase of unspecific antibodies induced by other antigens. Serological investigations represent a valuable tool to record the intensity and development in time of the Salmonella exposure in pigs farms. Examination of total Ig is an appropriate method to detect pig herds with a high level of Salmonella exposure, for detailed epidemiological studies in pig farms the examination of antibody isotypes will give more comprehensive information. PMID- 12894682 TI - [The epizootiology of tuberculosis of cattle in the Federal Republic of Germany]. AB - Mycobacterial strains from different outbreaks of tuberculosis of cattle in Germany from 1996 to 2001 were differentiated by two molecular biological methods (Spoligotyping, RFLP IS6110). The causative agent was in one case Mycobacterium (M.) africanum, in 10 cases M. bovis and in 17 cases M. bovis ssp. caprae, respectively. The results of the molecular biological methods are discussed from the perspective of epizootiology and the particular importance of infections by M. bovis ssp. caprae emphasized. Direct contact of the animals, purchase from infected stocks, infected zoo animals and wildlife, as well as livestock handlers are discussed as possible sources of infection. PMID- 12894683 TI - [Intracellular survival of Salmonella strains in Caco-2 cells]. AB - In the in vitro model using Caco-2 cells at different stages of differentiation the invasion and intracellular survival of virulent (predominant infection strains) and less virulent (predominant attenuated mutant strains) Salmonella strains were studied. The statistical evaluation of experimental data has shown that the logarithmized colony forming unit after 18 hours of incubation in differentiated cells (14 days old) is a suitable parameter for the determination of intracellular survival. Using this parameter a relationship between intracellular survival and Salmonella virulence (LD50 mouse) was demonstrated and quantified. The model presented could be suitable for the replacement of animal experiments after further investigations. PMID- 12894684 TI - [Baiting intervals and duration of control of the small fox tapeworm: a simulation study]. AB - Field trials to control the small fox tapeworm Echinococcus multilocularis failed to reduce the prevalence of the parasite persistently. With the experience from this empirical work, a simulation model for the population dynamics of the small fox tapeworm was developed. This model is used to examine the effect of different control strategies. Sufficient control duration is able to eradicate the parasite in simulation experiments. Baiting intervals of 4 or 6 weeks were found to be more efficient than shorter or longer intervals. The strategy used in field trials showed no sustainable effect in any of the simulation experiments. The applicability of simulation models for the planning of control measures for wildlife diseases is discussed. PMID- 12894685 TI - [Seroepidemiological studies of zoonotic infections in hunters in southeastern Austria--prevalences, risk factors, and preventive methods]. AB - The aim of this study was to investigate the seroprevalences to zoonotic pathogens in hunters, to propose preventive measures and to obtain more information about the occurrence of zoonotic pathogens in local wild animal populations. From 146 male and 3 female hunters originating from the south eastern Austrian federal states of Styria and Burgenland blood samples were taken and anamnestic data were obtained using a questionnaire. The serological investigations included the following viral, bacterial and parasitic zoonotic agents or zoonoses, respectively (antibody-seroprevalences in brackets): encephalomyocarditis virus (EMCV, 15%), Puumala-Hantavirus (10%), Newcastle Disease virus (NDV, 4%), borreliosis (IgG 42%, IgM 7%), brucellosis (1%), chlamydiosis (3%), ehrlichiosis (IgG 15%, IgM 3%), leptospirosis (10%), tularaemia (3%), Q fever (0%), Echinococcus multilocularis/E. granulosus (5%/11%), toxocariasis (17%). Out of a control group of 50 persons (urban population, no hunters) only one person was found to be seropositive for Toxocara canis and NDV and four for EMCV, all other results were negative in the control group. The high seroprevalences especially to Borrelia burgdorferi s.l., Ehrlichia spp., Leptospira interrogans, E. granulosus, E. multilocularis, encephalomyocarditis virus and Puumala virus demonstrate that hunters are particularly exposed to zoonotic pathogens. It should also be noted that one hunter was seropositive for Brucella abortus and five exhibited antibodies to Francisella tularensis. In these cases, as well as in the cases of the 15 seropositives for Leptospira interrogans, the suspected source of infection may- besides rodents--also include wild boars and brown hares. The infections with NDV and Chlamydophila psittaci may be traced back to contact with certain species of birds (potential risk: aviaries). For Hantaviruses, rodents are considered to be the main source of human infections. PMID- 12894686 TI - The influence of omega-3 polyunsaturated fatty acids (omega-3 pufa) on lactobacilli adhesion to the intestinal mucosa and on immunity in gnotobiotic piglets. AB - The effect of application of omega-3 polyunsaturated fatty acids (omega-3 PUFA) on intestinal colonization by Lactobacillus paracasei and on cellular immunity has been investigated in gnotobiotic pigs. The administration of polyunsaturated fatty acids positively affected the adhesion of Lactobacillus paracasei to the jejunal mucosa of gnotobiotic piglets. When compared to the control group, the number of Lactobacillus paracasei adhering to the jejunal mucosa was by 12% higher in piglets of the experimental group (5.10 log 10/cm2 vs. 4.55 log 10/cm2). The respective counts of Lactobacillus paracasei adhering to the ileal and colonic mucosa of 28 day old gnotobiotic piglets reached 4.45 and 5.05 log 10/cm2 in group C and 4.44 and 4.95 log 10/cm2 in group E. Omega-3 PUFA supplementation increased the phagocytic activity of neutrophils by almost 100% on day 28 of life as well as the subpopulations of lymphocytes (CD8) in the peripheral blood of germ-free piglets on day 21 of life. Our results indicate that the action of probiotics in the gut may be modulated by dietary PUFA. The stimulatory effect of PUFA upon adhesion of lactobacilli could be used for enhancing the effectiveness of probiotics in inhibiting digestive tract pathogens. PMID- 12894687 TI - Effect of silicate minerals (zeolite, bentonite, kaolin, granite) on in vitro fermentation of amorphous cellulose, meadow hay, wheat straw and barley. AB - The objective of the present experiment was to determine the effects of addition of silicate minerals, zeolite (Z), bentonite (B), kaolin (K), granite (G) on the rumen fermentation parameters, total gas, methane, total and individual volatile fatty acids (VFA) and hydrogen recovery in rumen fluid inoculum from sheep. Different materials (0.25 g) meadow hay (MH), wheat straw (WS), barley (BA) and amorphous cellulose (AC) were used as substrates. Silicate minerals (0.1 g) were added to the fermentation bottles containing substrates and rumen fluid inoculum and incubated for 72 h in vitro. The gas production technique simulates fermentation in the rumen was used to determine fermentation parameters. The total gas production was significantly higher compared to control for MH plus B (MHB), MH plus G (MHG), WS plus Z (WSZ), WS plus B (WSB), WS plus K (WSK), WS plus G (WSG), AC plus B (ACB), AC plus G (ACG), BA plus Z (BAZ), BA plus B (BAB), BA plus K (BAK), BA plus granite (BAG). Significant differences of the methane production were found between the controls, WSG, BAB and BAK. The total VFA concentration was increased in ACG (83.1 mM). The acetate: propionate (A:P) ratio of the control and additives ranged between 3.1 and 3.6 for MH, 2.7 and 3.5 for WS, 1.6 and 1.8 for AC and 2.3 and 2.9 for BA. It was concluded that the silicate minerals had no appreciable effect on the methane production, however, they support the microbial metabolism by influencing (bentonite, granite) and slightly influencing (zeolite, kaolin) the rumen fermentation. PMID- 12894688 TI - [Concentrations of carnitine and further biochemical parameters in the blood serum of healthy dairy cows and cows with abomasal displacement (dislocatio abomasi) and puerperal disorders]. AB - It was the aim of this study to compare the concentrations of total (TC) and free L-carnitine (FC) in blood serum of different groups of lactating cows. The animals were allotted into three groups, a) control animals (N = 11), b) cows with abomasal displacement (AD) (N = 5) and c) cows with puerperal disorders (PD) (N = 5). TC and FC were measured with an radioenzymatic assay. Blood samples were collected from 5 to 0 d before parturition (a. p.) and from 0 to 28 d after parturition (p. p.). It was of interest to examine whether L-carnitine might be limiting under certain conditions of metabolic stress which are typical for high yielding lactating cows. Concentrations of TC and FC (mumol/l) in control cows before and after parturition were 10.0* and 8.6* and 6.0-8.9 and 3.7-4.9, respectively. The corresponding TC and FC values for cows with AD were 19.6* and 8.9* and 10.5-20.7 and 4.8-6.9, respectively. Cows with PD showed TC and FC concentrations a. p. and p. p. of 15.7* and 9.2* and 10.3-13.0 and 4.8-6.3, h other puerperal disorders PD respectively (* only one value). TC and FC concentrations in serum of normal cows were higher before than after parturition. Cows, which developed post partal puerperal disorders had a higher prepartum concentrations of TC and CE, on the other hand, serum concentrations of FC was lower. Post partum cows with metabolic disorders showed higher CE levels than control animals. Cows which developed DA had significantly higher concentrations of TC and CE in serum only on d 7 p.p. than animals with other PD. It appeared that increased fat mobilization was regularly associated with responsive increases of CE concentrations in blood serum. PMID- 12894689 TI - [Bilateral avulsion fracture (apophyseolysis) of the calcaneal tuber in a heifer]. AB - This case report describes the clinical, radiographic, ultrasonographic and histopathological findings of a six-month-old heifer, suffering from bilateral fracture of the calcaneal tuber caused by osteochondrosis. The young cattle was admitted to the clinic for the evaluation of a left hindlimb lameness, having persisted for several weeks. Orthopaedic examination revealed a highly stilted gait with a lameness of the left hind limb, so severe that the heifer was only able to put some weight on the tip of the toe. The calcaneal region on both hindlimbs showed a diffuse swelling, palpation being very painful. The radiological examination revealed a fracture of the calcaneal tuber (apophyseolysis) of both hindlimbs. PMID- 12894690 TI - [Analysis of the population structure of the Black Forest Draught Horse]. AB - Gene contributions of foreign populations as well as coefficients of inbreeding and relationship were evaluated in 699 Black Forest Draught horses of Baden Wurttemberg actually registered in the year 2002. Based on nearly complete 5 generation-pedigrees and after taking into account the remaining incompleteness, the mean coefficient of inbreeding for the total population was 6.5%. The recently by incrossing with different breeds newly established lines of stallions showed significantly lower mean coefficients of inbreeding. High rates of inbreeding of about 1.6% in the last five generations could also be faced by incrossing stallions of foreign coldblooded populations what resulted in a decrease of inbreeding in the last generation. In the total population the mean degree of relationship was 16%. The mean degree of relationships within lines of stallions was between 18.3 and 26.8%. The coefficients of relationships between lines of stallions varied between 5.1 and 16.6%. Especially, the newly established lines of stallions showed a lower mean degree of relationships to the other different lines of stallions. The proportion of purebred Black Forest Draught horses in the total population was nearly 70%. Assuming that most animals of unknown origin were purebred, the proportion of purebred Black Forest Draught horses reached about 90%. Austrian Noric, Swiss Freiberg and South German Coldblood stallions were the most important contributors to the Black Forest Draught horse population. PMID- 12894691 TI - [Chronic-idiopathic enteropathy in cats--a case control study]. AB - Aim of this study is to clinically, microbiologically and histopathologically characterize inflammatory bowel disease (IBD) in the cat. Nine cats with chronic persistent or intermitent vomitus and diarrhea were examined between 1998 and 2001. All cats had a thuorough diagnostic workup performed. Full thickness biopsies from stomach, duodenum, jejunum, ileum and colon were surgically obtained for histopathological examination. Duodenal juice was obtained by direct aspiration for microbiologic qualitative and semiquantitative examination. Seven cats euthanized for other medical reasons were used as controls. Six cats had a lymphoplasmacytic IBD and three an eosinophilic IBD. Four cats with IBD had additional diseases diagnosed. Three cats with IBD had elevated bacterial counts. Retrospectively no correlation could be found between clinical symptoms and histopathological results. Serum TLI was not able to differentiate chronic pancreatitis and IBD. Serum folic acid and cobalamin did not correlate with the distribution of lesions in the gastrointestinal tract. Finally, no correlation was found between bacterial counts in the proximal duodenum and IBD. PMID- 12894692 TI - [Gutteral pouch tympany in German warmblood foals: influence of sex, inbreeding and blood proportions of founding breeds as well as estimation of heritability]. AB - The objective of the present study was to analyse the importance of the influences of sex, inbreeding coefficient, proportion of genes of the original breeds and the additive genetic contribution to the occurrence of guttural pouch tympany in foals belonging to German Warmblood breeds. Foals affected by guttural pouch tympany were ascertained in the Clinic of Horses, School of Veterinary Medicine Hannover. This data set comprised 22 German Warmblood foals with guttural pouch tympany, which were patients of the Clinic for Horses between 1994 and 2001. Information on the pedigrees and all available relatives of these patients allowed us to group the affected foals into five families with a total of 289 animals. Female foals were significantly more often affected by guttural pouch tympany. The difference was 16.6% in favour of female foals. The size of the inbreeding coefficient was not important for the occurrence of guttural pouch tympany. The proportion of the genes of the breeds Arabian, Thoroughbred and Trakehner were not significantly different from a randomly selected sample of 10% of foals born in the same birth years and the same region. The heritability estimates for the frequency of guttural pouch tympany using a threshold model was 0.81 +/- 0.16. This is the first report that could show a genetic component responsible for guttural pouch tympany in horses. PMID- 12894693 TI - Group discussion that focuses the mind. PMID- 12894694 TI - Putting words into action: the JWC/Molnlycke Scholarship from the winner's perspective. PMID- 12894695 TI - Exploring the psychosocial concerns of outpatients with disfiguring conditions. AB - OBJECTIVE: This study was undertaken to establish the extent and type of psychosocial need in outpatients attending for treatment for a variety of disfiguring conditions. METHOD: A total of 220 outpatients receiving treatment for burns, skin conditions or head and neck cancer, together with those seeking plastic surgery for other appearance concerns, completed standardised measures of anxiety, depression, social anxiety and quality of life. Participants also completed a semi-structured interview exploring individual concerns and satisfaction with care. RESULTS: A considerable proportion of the outpatients with disfiguring conditions experienced psychosocial difficulties, displaying raised levels of anxiety, depression, social anxiety and social avoidance and reduced quality of life. Levels of psychosocial distress were not well predicted by the severity of disfigurement. CONCLUSION: The psychosocial needs of these patients are not well met in the outpatient setting. Recommendations are made on how patients' concerns might be more effectively addressed. PMID- 12894696 TI - Personal story offers insight into living with facial disfigurement. AB - This paper looks at the long-term effects of living with facial disfigurement caused by a burns injury. It applies a psychosocial model to understand how someone with such an injury confronts and adapts to living with a changed self image. PMID- 12894697 TI - The link between the peel force of adhesive dressings and subjective discomfort in volunteer subjects. AB - OBJECTIVE: The study compared the level of discomfort experienced by healthy volunteers on the removal of a range of adhesive wounds. METHOD: This was an open, within subject comparative study of six adhesive dressings in 24 volunteers. The test site was the lower back. Allocation of test materials to the test sites was randomised. The peel force of removal was recorded after 24 hours of application using a device that removed the dressing at a constant speed and angle to the skin surface. The discomfort experienced at each removal was assessed by the subjects themselves using an electronic visual analogue scale. RESULTS: Overall, Mepilex Border was given a significantly lower discomfort score (p < or = 0.01) by the subjects than the other dressings. There were no clear differences between the five other products tested. Tielle and Allevyn Adhesive had significantly higher (p < or = 0.05) peel force than the other products. Mepilex Border caused less discomfort on removal than Duoderm Extra Thin, Biatain and Versiva, even though the peel force was similar. Tielle and Allevyn had higher peel force, but the levels of discomfort were not significantly higher for these products. CONCLUSION: It may be that the level of discomfort experienced by subjects on removal of an adhesive dressing is not entirely dependent on the peel force and that other aspects of the interaction of the skin surface and adhesive play a role. PMID- 12894698 TI - Sepsis: its causes and effects. AB - Sepsis contracted after malignancy, surgery or trauma is caused by a variety of micro-organisms ranging from viruses and fungi to bacteria. If untreated, it can lead to permanent damage to organs and tissues, or even to the patient's death. PMID- 12894699 TI - How sedative medication in older people affects patient risk factors for developing pressure ulcers. AB - OBJECTIVE: This US study set out to examine the relationship between pressure ulcer risk and sedation. The researchers examined the frequency of sedative use and the severity of pressure ulceration among older patients hospitalised for skin-ulcer treatment. They compared 91 patients who had been sedated before admission to hospital with 101 who had not. METHOD: A retrospective chart-review study was carried out between August 1994 and September 2001 in a tertiary-care metropolitan teaching hospital in the US. A total of 278 patients were identified from computerised discharge records. They were aged 60 years or more and had been discharged with medically treated skin ulcers, skin grafts, debridements or cellulitis. They had a principal or secondary diagnosis of chronic skin ulceration. RESULTS: Of the patients with pressure ulcers, 45.5% had been on sedation before admission. They were only slightly more likely to have come from nursing homes and there were no significant differences in prevalence of conditions such as oncological diseases or spinal-cord injury between them and non-sedated patients. However, patients sedated before admission were more likely to be female (67.1%, p = 0.04) and had almost a fivefold higher incidence of extremely severe ulceration (p < 0.0001). CONCLUSION: Almost half of the older patients hospitalised with pressure ulcers had been taking sedatives before admission. They were more likely to have extremely severe ulcers with necrotic tissue, pressure ulcers in multiple sites, and the largest and deepest ulcers. PMID- 12894700 TI - [Recent progress in the treatment of acute leukemia]. AB - Acute leukemia is classified broadly as either acute myelogenous leukemia (AML) or acute lymphoblastic leukemia (ALL). The main treatments remain remission induction therapy and postremission chemotherapy. The advances in chemotherapy for pediatric patients with ALL have been dramatic, with some 95% achieving complete remission, and long-term survival is 60-80%. Among adults, high long term survival rates due to improvements in chemotherapy for B-cell type ALL and core binding factor leukemias have been reported. For adult leukemias overall, however, long-term survival rates have stalled at 15-40% despite the high remission rate attained. In most cases this is due to a recurrence. Among the prognostic factors reported for acute leukemia, chromosome type may be cited as the currently most reliable. Acute leukemia patients are classified based on chromosome type, and the postremission therapeutic strategy is considered in terms of an appropriate combination of chemotherapy and hematopoietic stem cell transplant. This accounts for an important part of the treatment given today. Target-based therapies such as all-trans-retinoic acid (ATRA) for AML have brought dramatic improvements in treatment results. The effect of imanitib against Philadelphia chromosome positive ALL, for which the prognosis is poor, is also attracting attention. Moreover, promising new treatment strategies that have been developed, including cord blood transplant, mini-transplant, antibody therapy, and immunotherapy, Clinical studies of PCR and other means to reveal small residual lesions and estimate prognosis are also making progress. In the future it will be possible to identify prognostic factors in genetic tests such as DNA microarrays and single nucleotide polymorphisms, so that the optimum treatment can be selected for individual patients. PMID- 12894701 TI - [Phytoestrogen and cancer prevention]. AB - Phytoestrogens are defined to be plant chemicals that modify estrogenic effects in the body by binding to the estrogen receptors in mammals. Isoflavones, coumestane, lignan, and prenylflavones are examples of these, with isoflavones from soy foods and lignans from rye being a major dietary contribution. Mechanisms of cancer prevention by these phytoestrogens are reviewed, and human epidemiological studies, especially for breast and prostate cancers, are summarized and the results discussed. PMID- 12894702 TI - [Progress of diagnosis for esophageal cancer]. AB - Although the possible existence of micrometastasis in superficial esophageal cancer cases is the most important factor in deciding the therapeutic strategy, it is difficult. There are also limits to the diagnosis of the depth of tumor invasion by endoscopy and EUS. Therefore, the extension of the indication of EMR is planned. Then, the complement of diagnostic ability by EMR is being tried. Sentinel lymph node (s) identification and its biopsy also seem to be useful. In advanced esophageal cancer cases, diagnostic accuracy has been improving by speedup of CT and MRI scans and various image processing technology. More accurate diagnosis of metastasis using PET is much awaited. However, there is still insufficient ability to detect micrometastasis to date. Molecular bio markers with comprehensiveness are not a valuable selection at this stage. Although advanced equipment has brought more diagnostic accuracy, ways to integrate or use these tools at the clinical level are sought. Some equipment developing and how to integrate them and how to select the appropriate examination in each patient are problems which are required in clinical practitioners. PMID- 12894703 TI - [Endoscopic mucosal resection for esophageal cancer]. AB - Lymph node metastasis or microvascular permeation is rare among esophageal cancer which remains within the epithelium or the lamina propria mucosae. Endoscopic mucosal resection (EMR) is recommended for them as a radical treatment. Radical esophagectomy had been indicated for esophageal cancer reaching to the muscularis mucosae for their incidence of lymph node metastasis (10%). Recently, number of m3 or sm1 cancer cases treated by EMR has been increased, for some clinical trial succeeded to show that there is no significant difference between the prognosis of patient treated by EMR and by surgery. Thirty one patients (14%) have been lost among 219 patients who underwent EMR (mucosal cancer: 196 and submucosal cancer: 23). Four cases (13% of all patients lost after EMR) died of esophageal cancer, (one case with m3 cancer was lost by lymph node metastasis, one with sm2 cancer by liver metastasis, and two patients who refused surgical treatment by local recurrence after EMR), nineteen by other diseases than esophageal cancer and eight by cancer at other organs (lung: 3, hypoharynx: 2, mesopharynx: 1, stomach: 1, ureter: 1). Three of them were found concomitantly and 5 metachronously. Local recurrence was found in 8.3% of all patients treated by EMR. All patients with recurrence had received piecemeal resection. Sixty seven percents of all lesion of local recurrence was detected by endoscopic surveillance within one year after EMR. All recurred lesions were treated by EMR and pathological studies on resected specimens revealed that all recurred lesions were mucosal cancer. Metachronous esophageal cancer was found in 11% of all EMR cases. Sixty five percent of all metachronous cancer were detected in one to three years after EMR. Metachronous esophageal cancer after EMR was frequently found among cases with esophageal mucosa which has many small unstained areas. Malignant lesions were found in 33 cases (15%) of all patients treated by EMR synchronously and 37 (17%) metachronously. The stomach and the head and neck are most frequent site of associated cancers. PMID- 12894704 TI - [Review of endoscopic surgery for esophageal cancer]. AB - Our treatment strategy for esophageal cancer is as follows; mucosal cancer with the minimal risk of lymph node metastasis is treated by endoscopic mucosal resection (EMR). Submucosal cancer and more invasive lesions are surgically treated by thoraco- and aparoscopy-assisted esophagectomy. In May 1997, the first case received totally endoscopic esophagectomy, and which is survival with no tumor recurrence. Fifty consecutive cases successfully received endoscopic esophagectomy. This surgery should be performed as one major component of the multi-disciplinary treatment. PMID- 12894705 TI - [Minimally invasive esophagectomy with 10 cm thoracotomy assisted thoracoscopy for the thoracic esophageal cancer]. AB - We have been successfully performing minimally invasive esophagectomy through a 10-cm thoracotomy and two trocar ports. Esophagectomy can be performed safely and efficiently via the thoracoscopy. We have also adapted hand assisted laparoscopic surgery (HALS) for the abdominal procedure through a 7-cm laparotomy and three trocar ports. One of the important points of esophagectomy for thoracic esophageal cancer is performing lymphadenectomy near either side recurrent laryngeal nerve. Another important point is to preserve the bronchial branch of the vagus nerve and both bronchial arteries if they are without cancerous invasion. We also preserve the azygos vein and the thoracic duct to minimize surgical invasion. Standard thoracotomy and laparotomy are significantly invasive procedures with potential complications and prolonged healing; minimally invasive esophagectomy has the potential to minimize morbidity and decrease healing time. PMID- 12894706 TI - [The management with clinical path for esophagectomy for esophageal cancer]. AB - We have introduced a clinical path for esophagectomy of esophageal cancer from January, 2001 and got good results. It is important of the adaptation and the evaluation of variances for the critical path. Patients and co-medicals understood deeply the disease and the treatment of esophageal cancer. We could improvement the process of treatments by changing the critical path easily. We concluded that a critical path was adapted for esophagectomy of esophageal cancer, and it will be getting important. PMID- 12894707 TI - [Chemoradiotherapy for esophageal cancer]. AB - Since mucosal (T1a) esophageal cancer is well controlled by endoscopic treatment, chemoradiotherapy (CRTx) is not indicated. However, for a submucosal (T1b, N0) esophageal cancer, CRTx may be the first line of treatment, since it can provide a good response rate, with an excellent survival rate comparable to that after esophagectomy. Definitive CRTx is also in the first line of treatment for a T4 esophageal cancer, because there was no difference in the survival rate between CRTx with surgery and CRTx without surgery in our trial. Esophagectomy is indicated only for non-responders or recurrence-salvage surgery. For patients with a potentially-resectable (T2-T3) esophageal cancer, esophagectomy offered a longer survival rate than CRTx did, in our series. However, there remains controversy over the efficacy of CRTx for a T2-T3 esophageal cancer. It has been reported by the National Cancer Center Hospital East Group that definitive CRTx provided the same survival rate as esophagectomy. A prospective trial comparing the survival rate after esophagectomy and that after CRTx for a T2-T3 esophageal cancer is needed. PMID- 12894708 TI - [Progress of gene therapy for esophageal cancer in Japan]. AB - Retrovirally expressed interleukin-2 gene, granulocyte macrophage-colony stimulating factor gene, herpes simplex virus-thymidine kinase gene and p53 gene in human esophageal cancer cells showed antitumor effects in a nude mice xenotransplant model. We established a clinical protocol of gene therapy for advanced esophageal cancer using the wild type p53 gene with an adenovirus vector. In December of 2000, we began the first tumor suppressor gene therapy trial. Now, this trial, which has 9 patients. There have been no serious adverse event excluding fever and local pain. The feasibility of this treatment appears fairly good in these 9 cases. Furthermore, we developed a new method for transducing genes without a virus vector since a virus vector has several potentially unwanted properties. In vivo electroporation is a useful strategy for cancer gene therapy. Moreover, electric pulse to established solid tumors increases intracellular concentrations of chemotherapeutic agents. Transduction of the wild-type p53 gene by electroporation decreased the amount of nedaplatin required for tumor suppression. Electrochemo-gene therapy is a relatively simple method and can produce a better therapeutic effect. PMID- 12894709 TI - [Combination chemotherapy with nedaplatin and 5-fluorouracil for oral squamous cell carcinomas]. AB - We used a new combination chemotherapy with nedaplatin (CDGP) and 5-fluorouracil (5-FU) in eleven fresh patients with oral squamous cell carcinomas. 5-FU was administered at a dose of 1,000 mg/body by continuous infusion for 24 hours on days 1 to 5. CDGP was administered at a dose of 80 or 100 mg/m2 by drip infusion for 120 minutes on day 5. The response rates of total (1- or 2-course) and 2 course group were 54. 5% and 83.3%, respectively. Adverse drug reactions were limited to two cases of grade 3 toxicity with anorexia. The combination chemotherapy with 5-FU and CDGP in place of cisplatin and 5-FU seemed to play an important role as neo-adjuvant chemotherapy for oral squamous cell carcinomas. PMID- 12894710 TI - [UFT plus cisplatin with concurrent radiotherapy in unresectable stage III non small cell lung cancer--its application to outpatient practice]. AB - BACKGROUND: Combination chemotherapy with UFT and cisplatin is active and less toxic for advanced non-small cell lung cancer. This treatment is likely to be applied to concurrent chemoradiotherapy for locally advanced non-small cell lung cancer, especially in the outpatient setting. PATIENTS AND METHODS: Ten patients with unresectable stage III non-small cell lung cancer received the UFT plus cisplatin treatment combined with concurrent radiotherapy. The chemotherapeutic regimen consisted of oral administration of UFT 400 mg/m2 daily and venous infusion of cisplatin 20-25 mg/m2 on days 8-10. The administration of cisplatin was repeated every 3-4 weeks. Thoracic radiation started on day 8, and was completed to a total dose of 60-70 Gy. RESULTS: Adverse events (grade 3 or 4) occurred in 2 patients (esophagitis 2, leukopenia/neutropenia 1) with no treatment-related death. There were 7 partial responses (response rate 70.0%; 95% C.I., 41.6-98.4%). The median survival time was 18.7 months with a 1-year survival rate of 77.8%. Two patients uneventfully received the treatment in an outpatient setting. CONCLUSION: With regard to the quality of life of patients, UFT plus cisplatin with concurrent radiotherapy might be the treatment of choice for unresectable stage III non-small cell lung cancer. PMID- 12894711 TI - [A clinical results of TS-1 in advanced and recurrent gastric cancer in our hospital]. AB - A total of 40 patients with advanced and recurrent gastric cancer in our hospital were treated with TS-1 alone, and the efficacy of treatment, survival time, and adverse effects were examined. TS-1 was administered with the usual dosage and dose regimen. Response to treatment included a complete response (CR) in 3 cases, partial response (PR) in 8 cases, no change (NC) in 10 cases, and progressive disease (PD) in 7 cases. The response rate was 39.3%, and among the 28 patients with evaluable lesions TS-1 produced a high response rate of 56.3% in 16 patients who had undergone prior therapy. The median survival time (MST) was 478 days in the 28 patients with evaluable lesions, excluding patients with peritoneal dissemination, and 283 days in the 12 patients with peritoneal dissemination. The outcome was markedly poorer in the patients with peritoneal dissemination than in the patients with evaluable lesions. The incidence of grade 3 or higher adverse effects was 20%, including two cases in which decreased dihydropyrimidine dehydrogenase (DPD) activity was suspected, and one case in which decreased dihydropyrimidinase (DHP) was suspected. Although the effect of TS-1 alone on gastric cancer is significantly superior to that of any conventional cancer drugs, the results of this study suggest that the antitumor effect varies with the site of the target lesions and according to whether the lesion is a remnant or recurrence. PMID- 12894712 TI - [Multi-center trial of gemcitabine for 49 patients with advanced pancreatic cancer]. AB - Forty-nine patients with unresectable pancreatic cancer (stage IV disease) received gemcitabine in a multi-center trial in the Fukuoka pancreatic cancer chemotherapy group, Japan. No complete remissions, 5 partial remissions (10%) and 25 no changes (51%) were obtained. Gemcitabine could maintain QOL. Main toxicities were hematologic, especially neutropenia. Neutropenia tended to appear in early administration. Non-hematologic toxicities were anorexia, nausea/vomiting, and skin rash. The mean overall survival period was 7.5 months. Carcinomatous ascites and/or pleural effusion resulted in a poor prognosis (average survival 3.1 months). Gemcitabine could be given without severe toxicities in outpatient clinics. These results suggested that gemcitabine is currently a first-line therapeutic agent for advanced pancreatic cancer. PMID- 12894713 TI - [Alveolar hemorrhage as a possible adverse drug reaction by gefitinib (ZD1839, Iressa)]. AB - We experienced a case in which severe alveolar hemorrhage occurred in the course of gefitinib therapy. A 56-year-old man with non-small cell lung cancer had been treated with CDDP + CPT-11, CDDP + GEM + VNR, CDDP + TXT. After the chemotherapy with these regimens was found to be ineffective, daily oral gefitinib was started. Four weeks later, the patient complained of cough, bloody sputum and dyspnea. Chest X-ray and CT showed bilateral infiltrations with air bronchogram. Fiberoptic bronchoscopy revealed alveolar hemorrhage with an increase of lymphocytes in the BALF. After the cessation of gefitinib therapy and the administration of steroid, he gradually recovered. PMID- 12894714 TI - [A case of recurrent lung cancer successfully treated with vinorelbine and cisplatin/carboplatin]. AB - A 73-year-old male underwent lobectomy with ND2a lymph node dissection and resection of the superior vena cava for right lung cancer in December 1998 at Akita University Hospital. Histopathological examination demonstrated moderately differentiated adenocarcinoma (pT4 (SVC) N2M0, Stage IIIB). He received 1 course of a combination of cisplatin (CDDP) and etoposide (ETP) as postoperative adjuvant therapy. In March 2001, he again underwent partial resection of the right lung (S8) due to recurrence. In December 2001, a new left lung metastatic tumor was found. The patient was transferred to our hospital, where he was given 1 course of vinorelbine (NVB) 25 mg/m2 (day 1, 8) and CDDP 80 mg/m2 (day 1). Subsequently, he received 2 courses of vinorelbine (NVB) 25 mg/m2 (day 1, 8) and carboplatin (CBDCA) 430 mg/body (day 1). After the chemotherapy, a complete response (CR) of metastatic lesions was achieved. Adverse reactions were grade 3 neutropenia, grade 2 alopecia and grade 1 nausea/vomiting. The combination of vinorelbine and platinum agent (CDDP/CBDCA) is a useful regimen in treating recurrent non-small-cell lung cancer. PMID- 12894715 TI - [A case of postoperative recurrent breast cancer with multiple lung metastases that completely responded to combination therapy of docetaxel (TXT) and medroxyprogesterone acetate (MPA)]. AB - A 54-year-old female had undergone surgery for breast cancer 5 years ago, after which she developed multiple lung metastases, in spite of treatment with various postoperative chemoendocrine therapies. The patient who had recurrent breast cancer with multiple lung metastases was treated with a combination of docetaxel (TXT) 80 mg/body three cycles, two courses of 40 mg/body four times and medroxyprogesterone acetate (MPA) 600 mg p.o. daily. Six months later, the lung metastases had completely disappeared on chest CT-scan. Complete remission has been maintained for one and half years. The use of combined chemoendocrine therapy with TXT and MPA is considered effective for recurrent breast cancer as second-line therapy. PMID- 12894716 TI - [Effective combination chemotherapy using weekly trastuzumab and paclitaxel in the treatment of a recurrent breast cancer patient with liver metastases]. AB - A 48-year-old woman was diagnosed with multiple locoregional recurrence five months after receiving breast conserving surgery, and she underwent mastectomy. CEF therapy was combined following surgery; however, lung metastases developed 9 months after the initial surgery. A CR has been observed after monthly use of docetaxel alone for 14 months. Since multiple liver metastases were detected 17 months later, the patient was given monthly docetaxel infusions for 2 months followed by weekly trastuzumab infusions for 8 weeks, resulting in PD. Thereafter, combination of weekly trastuzumab with paclitaxel was used and a nearly CR remained for 10 months. Although multiple brain metastases were seen 36 months later, vertigo was mostly controlled with whole brain radiotherapy. Combination of weekly trastuzumab and paclitaxel therapy is effective for liver metastases from breast cancer and useful considering the slight side effects and possibility of long-term administration. PMID- 12894717 TI - [Four cases of gastric cancer with multiple hepatic metastases successfully treated with TS-1 in combination with low-dose cisplatinum]. AB - We treated five cases for multiple hepatic metastases from gastric cancer with a novel combination of TS-1 and low-dose cisplatinum (CDDP). TS-1 was orally administered at 100 mg/body/day every day or only on weekdays, and 10 mg of CDDP was infused once or twice a week. The efficacy was evaluated by body CT after the treatment. The CT showed more than a 60% reduction of hepatic tumors in four patients. The tumor markers, CEA and CA19-9, were reduced to 10%. The response rate was 80%. Adverse reactions of grade 1 anemia were observed in two patients and grade 1 leucopenia in one patient. The liver function normalized in one patient. The hemoglobin level was increased from 6.8 g/dl to 11.8 g/dl in one patient. In conclusion, this combined chemotherapy of TS-1 and low-dose CDDP proved useful for advanced gastric cancer patients with multiple hepatic metastases, in view of its therapeutic efficacy, patients' quality of life and low toxicity. PMID- 12894718 TI - [A case of small cell carcinoma of the stomach]. AB - We report a case of small cell carcinoma of the stomach in a 60-year-old man. The patient was found to have a gastric tumor by routine upper gastrointestinal endoscopy for chronic gastrititis, and was admitted to the hospital. The patient underwent a curative distal gastrectomy with D2 lymph node dissection following a diagnosis of early gastric cancer at the posterior wall of the antrum, in June 2001. Surgical findings were T1, N1, P0, H0, M0, stage I b. The final pathological diagnosis was small cell carcinoma of the stomach with severe lymphatic and vascular invasion (INF alpha, sm, ly3 v2, n1 (+)). In a review of 47 cases seen in the Japanese literature, only 5 patients had early gastric cancer. Lymph node metastasis was observed in 95.0% of all cases, and liver metastasis in 41.0%. Four of the 5 patients with early gastric cancer (80.0%) had lymph node metastases. The prognosis was reported to be extremely poor because small cell carcinoma frequently metastasizes to lymph nodes and/or the liver even in an early stage. The chemotherapy selected by many hospitals is mainly CDDP, but the use of chemotherapeutic drugs varied from hospital to hospital and no effective chemotherapy for the disease has been established as yet. PMID- 12894719 TI - [A case of advanced gastric carcinoma successfully treated with TS-1/CDDP as a neoadjuvant chemotherapy]. AB - We report a patient with gastric carcinoma successfully treated with TS-1/CDDP as a neoadjuvant chemotherapy. The patient was a 72-year-old man who had suffered from general malaise with severe anemia and was diagnosed as type 2 carcinoma, having bulky N2, with suspected invasion of the pancreas. TS-1 (100 mg/day) was administered orally every day for 21 days followed by 14 days rest, and CDDP (20 mg/body) was administered by intravenous infusion at day 8, as one course. He was treated as an outpatient, and two courses of treatment resulted in a marked reduction of the lymphnode metastasis without toxicity. Subsequently, he underwent curative surgery consisting of total gastrectomy with D2 lymph node dissection. TS-1/CDDP therapy is very useful as a neoadjuvant chemotherapy especially for an outpatient until surgery. PMID- 12894720 TI - [A case report of complete remission of liver metastases from colorectal cancer treated with continuous hepatic arterial infusion]. AB - We encountered a patient with liver metastases from colorectal cancer in whom continuous hepatic arterial infusion brought complete remission. A 58-year-old man was admitted to our hospital for advanced rectal cancer with multiple liver metastases. He underwent a low anterior resection (D2). Continuous hepatic arterial infusion of 5-FU (250 mg/day) with a weekly arterial infusion of MMC (4 mg) was performed for 14 days. Six continuous hepatic arterial infusions resulted in a complete remission. The patient has been free from any sign of recurrence for 37 months after the operation. Continuous hepatic arterial infusion using 5 FU and MMC seems to be effective in the treatment of multiple liver metastases from colorectal cancer. PMID- 12894721 TI - [A case of advanced gallbladder cancer with biliary tract stenosis which responded to TS-1 chemotherapy]. AB - TS-1 is reported to be beneficial for advanced gastric cancer, but there is no report on its use for advanced gallbladder cancer. The present patient was a 64 year-old woman with advanced gallbladder cancer with severe biliary tract stenosis. The primary tumor was located in the neck of the gallbladder and peripancreatic lymph node metastasis was detected. TS-1 100 mg/day was administered orally for 21 days and CDDP 30 mg/day on days 1, 8 by drip infusion. Grade 4 neutropenia was observed in the first cycle, and TS-1 alone was used for further treatment. After 2 courses, primary tumor showed PR and lymph node metastasis had disappeared. Biliary stenosis was remarkably improved. We conclude that TS-1 might be beneficial in the treatment of advanced gallbladder cancer. PMID- 12894722 TI - [Pharmacokinetics of paclitaxel and carboplatin in a hemodialysis patient with metastatic urothelial carcinoma--a case report]. AB - A 69-year-old-man undergoing hemodialysis for 8 years developed metastatic urothelial carcinoma and received combination chemotherapy with paclitaxel and carboplatin. Paclitaxel 175 mg/m2 was given as a 3-hour intravenous infusion, and carboplatin was dosed to the area under the plasma concentration-time curve (AUC) of 5 mg.min/ml calculated according to the Calvert formula as a 30-minute intravenous infusion immediately after paclitaxel. Hemodialysis was started 1 hour after carboplatin, then their pharmacokinetics was determined in the patient. The AUC of paclitaxel and carboplatin were 15.2 and 64.56 micrograms.h/ml, respectively. The metastatic tumor size was reduced by more than 20% after 3 courses of this chemotherapy. Grade 3 neutropenia and grade 1 thrombopenia were observed. This is the first report that the combination of paclitaxel and carboplatin is feasible in a patient with metastatic urothelial carcinoma undergoing hemodialysis, with low toxicity and safety. PMID- 12894723 TI - [Metastatic gastrointestinal stromal tumors responded to the treatment with STI571 after polysurgery for recurrent lesions--report of two cases]. AB - Two cases of metastatic gastrointestinal stromal tumors (GIST) that had responded to the treatment with STI571 were presented. Case 1 was a 49-year-old woman who had undergone proximal gastrectomy because of a giant submucosal tumor of the stomach. For 21 months after surgery, the patient received repeated tumor removal four times due to hepatic metastasis and/or peritoneal recurrence. Thereafter, the treatment with STI571 at a dose of 400 mg/day was initiated. Eight months after the administration, only a small hepatic metastasis was detected on a film of CT scan, and any signs of peritoneal recurrence were observed. Case 2 was a 61 year-old man who underwent emergency surgery for a retroperitoneal tumor that had caused massive intestinal hemorrhage resulting in critical shock. The patient underwent the surgery three times for recurrent lesions. Because further tumor removal had become nearly impossible, STI571 at a dose of 400 mg/day was administered 35 months after initial surgery. Six months after treatment the hepatic lesions were shrunk, but the number of retroperitoneal lesions increased. At present, the patient has no abdominal complaints and has a good quality of life. GIST was confirmed in both cases, by histopathological analyses of the resected specimens: positive expression of c-kit and CD34. These clinical observations suggest that ST1571 therapy for metastatic lesions from GIST may be preferred over aggressive, repeated tumor removal. PMID- 12894724 TI - [ImmuCyst intravesical (freeze-dried preparation made from the connaught strain of Bacillus Calmette Guerin) for the treatment of superficial bladder cancer and carcinoma in situ of urinary bladder]. AB - BCG has been used as a preventive vaccine of tuberculosis worldwide. However, in 1976, Morales et al. reported the effective use against superficial bladder cancer by intravesical administration. In Japan, in July, 1994, Rhone-Poulenc Rorer Japan Inc, started the cooperative clinical studies with Nippon Kayaku Co. Ltd. as an orphan drug. In the phase I/II studies, BCG preparation at 40.5, 81 and 121.5 mg was given intravesically slowly, keeping it for 2 hrs, in the bladder. This was repeated once weekly for 8 weeks. The CR rates were 60, 68.2 and 75% respectively, but there were no significant differences. The major adverse reactions were polyurea, urination pain, urination difficulty, hematurea and urinary leukocytosis which were frequently observed. Beside these, fever, malaise, loss of appetite and GPT elevation were also observed. In the late phase II study, in which 81 mg were given, CR rates in 39 response evaluable cases, was 61.5% (16/26), response rate (CR + PR) was 85.6% (22/26), and CR rate of CIS was 92.6% (12/13). There were responded cases in the recurred cases and those with previous chemotherapy, and the adverse reactions were polyurea, urination pain, urination difficulty, hematurea, malaise and loss of appetite mainly. CONCLUSION: Compared with anticancer drug treatments, BCG prevented the recurrence the disease for a long time. From these studies, freeze-dried BCG preparation (immu Cyst) seemed to be very useful for the treatment of superficial bladder cancer and carcinoma in situ of urinary bladder. PMID- 12894725 TI - [Oral chemolysis of uric acid stones]. AB - Oral chemolysis of uric acid stones was performed retrospectively. Twenty-one patients with upper urinary uric acid stones were given alkaline citrate (Uralyt U) orally. In the case of hyperuicemia, allopirinol was combined. In 11 out of 15 patients (73.3%) treated with oral chemolysis alone, stones were dissolved. In 4 out of 6 cases (66.7%) combined with extracorporeal shock-wave lithotripsy, administration of alkaline citrate shortened the period to be stone-free. In conclusion, we successfully treated 15 out of 21 cases (71.4%) with the administration of alkaline citrate. PMID- 12894726 TI - [Clinical study of chemoradiotherapy using low dose cisplatinum as X-ray intensifier in patients with urothelial carcinoma]. AB - Chemoradiotherapy using cisplatinum (CDDP) as the X-ray intensifier was performed on patients with urothelial carcinoma. Ten lesions in 9 patients, 6 patients with postoperative relapse and 3 who received the therapy as a palliative treatment for progressive carcinoma, were evaluated. Four of the patients with postoperative relapse had undergone adjuvant chemotherapy. On the day of the treatment, the 9 patients were given continuous intravenous infusion of CDDP at the dose level of 5-12 mg/day prior to external irradiation at 50-66 Gy. The response to the therapy was categorized as complete response in 5 patients, partial response in 4, and no change in 1. The response rate was 90%, indicating achievement of a good local control. Pain relief and improvement of hydronephrosis were also observed in patients who underwent the therapy for treatment of progressive carcinoma. All adverse reactions were mild in intensity. These results suggest that the chemoradiotherapy is useful for both patient groups, those who have a postoperative relapse and those who undergo the therapy as a palliative treatment for progressive carcinoma. PMID- 12894727 TI - [Clinico-statistical analysis of renal cell carcinomas in patients on hemodialysis]. AB - Eleven patients on hemodialysis that were surgically treated for renal cell carcinomas during the recent 10 years at our institutes were clinically analyzed. Patients' ages at presentation ranged from 35 to 70 years with an average of 54.8 years. Nine of the 11 patients were males and 2 were females. Periods between the introduction of hemodialysis and the presentation ranged from 1 to 21 years with an average of 11.7 years. The most frequent cause of hemodialysis was chronic glomerulonephritis. Five patients presented with macroscopic hematuria, which was the most frequent clinical manifestation. Transperitoneal nephrectomy through a lumbar oblique incision was performed in 9 of 12 surgical procedures. Transperitoneal resection and retroperitoneal endoscopic resection were performed on 2 patients and 1 patient, respectively. Blood transfusion was performed on 2 patients with retroperitoneal hemorrhage before or after operation and 2 patients with pre-existing renal anemia. Pathologically, 9 patients had pT1a disease. Patients were followed up for up to 7 years and 11 months. One patient died of the disease and 2 patients died of unknown causes. In conclusion, surgical removal of renal cell carcinomas was well tolerated, safe and effective treatment in patients under hemodialysis. PMID- 12894728 TI - [A case of urinary and miliary tuberculosis detected during examination for renal failure]. AB - A case of urinary and miliary tuberculosis in a 51-year-old male is reported. The patient with the complaint of low grade fever at night and appetite loss had visited another clinic. Laboratory data showed renal failure (serum creatinine 3.9 mg/dl, BUN 35.1 mg/dl) and he was referred to our hospital. Ultrasonography and computed tomography (CT) revealed bilateral hydronephrosis. Therefore we suspected postrenal renal failure. We performed cystoscopy in order to perform retrograde pyelography and to indwell double J stent. The bladder showed yellow whitish nodules all over the bladder. Tuberculous bacilli were detected in the urine by smear, polymerase chain reaction (PCR) and culture. Chest radiography and CT revealed multiple granular nodules and so diagnosed miliary tuberculosis. Immediately combination chemotherapy of with isonicotinic acid hydrozide, rifapicin, ethanbutal and pirazinamide was administered. Histopathology by bladder biopsy showed epithelioid cell granuloma. Chemotherapy was effective, but the vesical capacity was contracted to less than 50 ml, and we suspect that the patient will need vesical augmentation. PMID- 12894729 TI - [A case report: renal arteriovenous malformation associated with horseshoe kidney detected by hemorrhage due to injury]. AB - A case report of a patient with renal arteriovenous malformation associated with horseshoe kidney detected by hemorrhage due to injury. A 71-year-old man was injured in a traffic accident and conveyed to our hospital. The computed tomographic scan showed renal injury of the horseshoe kidney with retroperitonial hemorrhage. We treated him conservatively, but he complained of abdominal fullness and went into hemorrhagic shock. So, we performed arterial angiography. It showed a right renal arteriovenous malformation from which leakage of contrast medium was detected. Hemostasis was possible by embolization of the abnormal artery. He has not had any recurrence of renal arteriovenous malformation. To our knowledge, this case is the third report of renal arteriovenous malformation associated with horseshoe kidney in Japan. PMID- 12894730 TI - [Renal papillary necrosis managed by transurethral procedures: a case report]. AB - The patient was a 66-year-old female who had been commonly using an analgesic for rheumatism from age 40. She visited our hospital with the complaints of fever up and right flank pain. Right hydronephrosis and renal failure were pointed out, and she was referred to the urologic clinic. Retrograde pyelography showed a clubbed upper calyx and filling defect in the lower ureter. A ureter stent was positioned for drainage in the right ureter. Then her general state improved. Three weeks later, retrograde pyelography was performed again. Two filling defects were detected in the upper ureter. Since the obstruction persisted we observed the ureter by ureteroscopy. Two specimens black-brown in color and 8 mm in diameter were observed through the ureteroscope and were removed with a basket catheter. Histological examination of the specimens revealed necrotic transepithelial tissues. It was assumed that the tissues were derived from necrotic renal papilla. Four months later, a similar episode was observed in the left upper urinary tract. The same procedures were performed to manage the patient. In this case, drainage using a ureter stent was effective and conservative therapy was possible. This is the first reported case of renal papillary necrosis managed by transurethral procedures in Japan. PMID- 12894731 TI - [Retroperitoneal ganglioneuroma: a case report]. AB - A 67-year-old male presented for examination of a retroperitoneal tumor, incidentally found by abdominal computed tomography (CT). CT and magnetic resonance imaging (MRI) revealed a round heterogeneous tumor, 10 cm in diameter, at the left renal hilus and involving the left renal vein. The tumor was low intensity on T1-weighted MRI imaging, and high-intensity on T2-weight MRI imaging. The tumor was easily resected via a transabdominal approach. The pathological diagnosis was ganglioneuroma. PMID- 12894732 TI - [A case of lipoleiomyosarcoma of the spermatic cord]. AB - A 54-year-old male was referred to our hospital with the chief complaint of a painless inguinal mass. A hard mass was palpable beside the left spermatic cord, which was covered with a soft tissue mass. Computed tomographic scan showed an inguinal mass, which was well enhanced to vascular density. The operative appearance indicated a tumor in the left spermatic cord, so we performed a radical orchiectomy. Histopathological diagnosis was lipoleiomyosarcoma. Lipoleiomyosarcoma of the spermatic cord is quite rare, and our case was considered as the first report in Japan. PMID- 12894733 TI - [A case of primary adenocarcinoma of the epididymis]. AB - A 68-year-old man presented with right intrascrotal swelling. On palpation, a hard tumor without pain was recognized at the head of the right epididymis. Power Doppler ultrasonography revealed blood flow signals within the tumor. Surgical exploration was performed under the tentative diagnosis as possible malignant tumor of the epididymis. The right epididymis adhered to the testis so strongly, that the epididymis was resected with the testis. Pathological diagnosis was moderately differentiated adenocarcinoma of the epididymis. The results of general examinations on possible presence of primary lesions in other organs were all negative. Finally, the diagnosis of primary adenocarcinoma of the epididymis was obtained. He remains free of disease 17 months after surgery. PMID- 12894734 TI - [A case report: an urethral tumor with cavernous structure developed after optical urethrotomy]. AB - A 70-year-old man underwent transurethral resection of the prostate (TURP) in February 1999 and he received optical urethrotomy because of urethral stricture in May. In May 2000, a distal urethral tumor was found by urethroscopy. Endoscopic resection of the urethral tumor was performed. The tumor consisted of normal urethral epithelial tissue and had a cavernous structure. We conclude that the tumor was vegetation of the corpus spongiosum penis. PMID- 12894735 TI - [A case of intrascrotal rhabdomyosarcoma with para-aortic lymph node metastasis- complete response after multidrug chemotherapy]. AB - We report a case of intrascrotal rhabdomyosarcoma in a 59-year-old-male. The patient with a mass in the right scrotum was at first diagnosed with epididymitis and treated with antibiotics. The mass however, grew gradually and right orchiectomy revealed intrascrotal alveolar rhabdomyosarcoma. Para-aortic lymph node metastasis was pointed out by abdominal computed tomography. He received multidrug chemotherapy consisting of vincristine, actinomycin-D, cyclophosphamide, adriamycin, and cisplatin (IRS-III regimen 35). A complete response was obtained 3 months after the start of the chemotherapy. The patient is alive without recurrence 2 years after the surgery. PMID- 12894736 TI - [A case of small renal cell carcinoma (diameter 1.5 cm) presenting with a large amount of ascites due to peritoneal carcinomatosis]. AB - A 62-year-old man was referred to our department because of a small mass (1.5 cm in diameter) in the right kidney. His physical examination revealed a remarkably distended abdomen due to ascites. Paracentetic cytology of the ascites was suggestive of clear cell carcinoma. Under the diagnosis of renal cell carcinoma, interferon alpha and gamma were given subcutaneously and intraperitoneally, but his clinical status became worse gradually and he died 2 years after the diagnosis. An autopsy revealed papillary renal cell carcinoma. Ascites due to peritoneal carcinomatosis is a rare presentation, especially for such a small renal cell carcinoma. PMID- 12894737 TI - [A case of primary transitional cell carcinoma of the prostate]. AB - A 77-year-old man was referred to our hospital with a complaint of dysuria and right ischiodynia. He had had a hemi-thyroidectomy for thyroid cancer and right cervical lymphadenectomy three years and one year, respectively, before this visit. Prostate cancer was strongly suspected by transrectal examination with prostate specific antigen (PSA) elevated to 77.8 ng/ml. Pathological diagnosis of prostate biopsy specimen was transitional cell carcinoma with grade 3 malignancy and negative staining for PSA. Endoscopic examination showed a normal appearance of bladder and prostatic urethral epithelium. Urine cytology showed no malignant cells. However, immunostaining for PSA revealed that the cervical lymph node specimen resected before was moderately differentiated adenocarcinoma of prostate. He had multiple metastases to mediastinal and retroperitoneal lymph nodes and right ischium. Endocrine therapy (goserelin acetate depot, bicalutamide) and systemic chemotherapy (methotrexate, epirubicin, cisplatin) were performed combined with irradiation to right ischium metastasis. Two months later, he showed a complete response in PSA and partial response in lymph node metastases, but died of cancer 13 months later. PMID- 12894738 TI - [A case of spermatic cord tuberculosis]. AB - A 47-year-old man visited our hospital with a complaint of a right intrascrotal mass. The results of a laboratory examination were unremarkable. An ultrasonographic examination of the right scrotum demonstrated a low echoic legion, 2.5 x 1.3 x 0.7 cm in diameter. A right spermatic cord tumor was diagnosed. Right high orchiectomy was performed. Microscopic examination showed a granulomatous lesion with Langhans large cells. Tuberculin skin test was strongly positive. From these findings we diagnosed the patient with tuberculosis in the spermatic cord. PMID- 12894739 TI - [A case of unilateral giant hydronephrosis with contralateral dwarf kidney]. AB - A 54-year-old woman was admitted to our hospital for oliguria and left lower abdominal pain. She had renal dysfunction with a serum creatinine of 9.1 mg/dl and blood urea nitrogen of 96.5 mg/dl. Plain computed tomography and magnetic resonance imaging revealed right dwarf kidney and left giant hydronephrosis with extravasation of urine. MR-urography revealed left dilated ureter caused by ureterovesical junction (UVJ) stenosis. Therefore, percutaneous nephrostomy was immediately performed to treat postrenal failure, with resulting collection of approximately 1,650 ml of urine. Subsequently, left ureterocystoneostomy was performed for the treatment of UVJ stenosis because improvement of left UVJ stenosis had not been confirmed by nephrostography during follow-up. Judging from the past history of myoma operated and reactive fibrosis of stump of left ureter histopathologically, it was considered that acquired UVJ stenosis had led to giant hydronephrosis. PMID- 12894740 TI - [Can hospital transmission of adenoviral infections be controlled?]. PMID- 12894741 TI - [Transient dysfunction of the retinal ganglion cells after laser in situ keratomileusis]. AB - PURPOSE: To evaluate the influence of high intraocular pressure(IOP) on the retinal ganglion cell function induced by suction during laser in situ keratomileusis(LASIK) using Frequency Doubling Technology(FDT). METHODS: Twenty seven eyes of 15 patients were studied. Threshold values of 19 targeted areas, mean deviation(MD), and pattern standard deviation (PSD) were measured before LASIK, and 2 hours, 1 day, and 1 week after LASIK. These parameters obtained by N 30 program of FDT were compared between before and after LASIK. RESULTS: Significant decrease of MD and threshold value in all areas was recognized at 2 hours, but MD and threshold value in 6 of the 19 areas showed significant increase at 24 hours. Significant increase of PSD value at 2 hours was recognized. After 7 days, there were no significant differences in any parameters between before and after LASIK. CONCLUSION: These results indicated that a transient dysfunction of the magnocellular pathway might be induced by high IOP during LASIK procedure. PMID- 12894742 TI - [Age-related changes in the mucocutaneous junction and the conjunctivochalasis in the lower lid margins]. AB - PURPOSE: We investigated the age-related changes in the mucocutaneous junction and the conjunctivochalasis at the lower lid margin, and whether there is a correlation between the two or not. SUBJECT AND METHODS: Sixty eyes of sixty patients without ocular surface disorders were examined and they were divided into six groups by age (20-29, 30-39, 40-49, 50-59, 60-69 and 70-79 years old) and sex(five males and five females in each age group). The lower lid margins were divided into three parts(the temporal third, the middle third, and the nasal third) and the anteroplacement of the mucocutaneous junction and the conjunctivochalasis were graded 0, 1, 2 and 3. RESULT: In each part of the lower lid margins, there were significant age-related increases in both parameters(p < 0.005, p < 0.0001 respectively). Also, the grade of the anteroplacement of mucocutaneous junction increased with the grade of conjunctivochalasis. CONCLUSION: These findings suggest that there is a relationship between the anteroplacement of the mucocutaneous junction and the conjunctivochalasis with aging. PMID- 12894743 TI - [Subretinal washout for subtle subfoveal hard exudates in diabetic macular edema]. AB - PURPOSE: To evaluate the efficacy of subretinal washout for subtle subfoveal hard exudates in diabetic macular edema. METHODS: This study was done retrospectively on a series of patients with diffuse diabetic macular edema accompanied with subtle subfoveal hard exudates and operated on by one surgeon(NO). Patients ranged in age from 30 to 76 years(mean, 59 years). The postoperative follow-up interval ranged from 12 to 76 months, with a mean of 35 months. Two groups were identified. The first group contained all 26 eyes that had vitreous surgery with subretinal washout. This was compared with a second group of 51 eyes without subretinal washout. We compared the rate of occurrence of postoperative massive foveal hard exudates, visual acuity results, and complications with and without subretinal washout. RESULT: There was no significant difference in base line demographics between the two groups. Massive foveal hard exudates did not occur in eyes with subretinal washout, but occurred in 29 (57%) of the eyes without subretinal washout(p < 0.0001) and in 15 eyes which had undergone reoperation with subretinal washout. Visual acuity improved in 54% of the subretinal washout eyes and 45% of the eyes without it. Visual acuity improved to 20/30 or better in 23% of the eyes with subretinal washout and in 8% of the eyes without subretinal washout. There was no serious complication related to subretinal washout. CONCLUSION: Subretinal washout for subtle subfoveal hard exudates in diabetic macular edema may prevent massive subfoveal exudates and improve visual results. Further study is needed to investigate the pathogenesis. PMID- 12894744 TI - [Diurnal variation of intraocular pressure in normal-tension glaucoma]. AB - PURPOSE: Measurement of diurnal variation of intraocular pressure(IOP) is important for precise diagnosis of normal-tension glaucoma(NTG). We studied diurnal variation of IOP of NTG using a self-measuring tonometer. METHODS: A total of 159 patients(318 eyes) who were diagnosed as having NTG in Osaka Koseinenkin Hospital between 1994 and 2002 measured their own diurnal variation of IOP at home every 3 hours (8 times a day) using a prototype self-measuring non contact air-puff tonometer(Hometonometer). RESULTS: The maximum IOP, the minimum IOP, and the range of diurnal variation of IOP were 16.8 +/- 2.0 mmHg(mean +/- standard deviation), 12.8 +/- 1.7 mmHg, and 4.0 +/- 1.3 mmHg, respectively. Maximum IOP occurred most frequently at noon(24.3%), 9:00 am(21.4%), and 6:00 am(17.4%). In 69.2% of eyes, maximum IOP was found during outpatient clinic hours(9:00 am to 6:00 pm). Minimum IOP occurred most frequently at midnight(34.1%), 3:00 am(22.8%), and 9:00 pm(17.8%). CONCLUSIONS: Approximately 30% of NTG patients have maximum IOP outside of outpatient clinic hours, and therefore measuring IOP in the early morning is important for determining the precise diurnal variation of their IOP. We hope that a safe self-measuring tonometer with which patients can measure their own IOP will be come commercially available soon, so that we can provide them with more individualized glaucoma treatment using the appropriate combination of medicines. PMID- 12894745 TI - [Measuring accommodation time with a laptop computer]. AB - PURPOSE: Presently, there are no completely reliable examinations such as focus from a distant place to a near place during daily work. We developed a simple testing system for accommodation time using a 5 m chart and a laptop computer. MATERIAL AND METHODS: We studied 177 healthy peoples. They turned their eyes from the 5 m chart to the display of the personal computer at about 70 cm in front of the eyes. We measured the minimum time(MTC) for recognizing three Kanji(Chinese characters) randomly chosen from a total of 15 characters on the display. RESULT: The MTCs with + 1.25 D added were shorter than those with complete correction. The MTCs in both conditions extended with age. In subjects less than 40 years old, the MTCs measured with both eyes were significantly shorter than those with a single eye completely corrected. In subjects over 40 years old, there was no difference. The MTCs were shorter significantly with complete correction than with + 1.25 D added. The MTCs decreased as the accommodative amplitude increased. CONCLUSION: There is significant correlation between the accommodative amplitude and the MTCs. We can use the MTCs as one method of measuring time of accommodation. PMID- 12894746 TI - [Epidemic nosocomial keratoconjunctivitis caused by adenovirus type 4]. AB - PURPOSE: To evaluate the clinical features of nosocomial epidemic keratoconjunctivitis(EKC) occurring in the ophthalmology ward of Sapporo Medical University Hospital and to devise preventive measures for it. MATERIALS AND METHODS: We studied the symptoms and clinical course of 2 patients who had EKC and 16 patients who had EKC caused by nosocomial infections in our hospital. We attempted to detect adenovirus antigen and viral DNA from conjunctival swabs and also to isolate the virus. RESULTS: The clinical symptoms of EKC were conjunctival hyperemia in 18 patients(100%), conjunctival follicles in 11 patients (61.1%), discharge in 8 patients(44.4%), superficial punctate keratopathy in 7 patients(38.9%), swelling of the eyelids in 3 patients(16.7%), and fever in 3 patients(16.7%). 72% were positive for Adeno-check. Adenovirus type 4 was isolated from the conjunctival swabs. We considered that the route of hospital infection was infection from the doctor's hands, from eye drops, and from contact lenses. We disclosed that nosocomical EKC had occurred in our hospital. The hospital infection was eliminated by closing the ophthalmology ward and sterilizing instruments and washing hands. CONCLUSIONS: The delay of proper measures increased the risk of nosocomical infection. We recognized the importance of careful observation of patients and immediate preventive efforts in nosocomical infection. PMID- 12894747 TI - [Undergraduate education in ophthalmology]. PMID- 12894748 TI - [Results of a survey conducted among university hospitals on nosocomial adenovirus conjunctivitis]. PMID- 12894750 TI - Phenol removal in upgraded facultative waste stabilisation ponds. AB - Treatment of wastewater containing phenol using laboratory scale waste stabilisation ponds enriched with activated sludge was studied. After enrichment, the efficiency of these ponds under high phenol loading rates (i.e. up to 408 kg phenol ha(-1) day(-1)) was studied. At phenol loading rates of 6, 52, 312, 636 and 972 COD ha(-1) day(-1), the phenol removal efficiencies were 77, 69, 76, 59 and 52%, respectively. The results suggest that enrichment is an effective potential method to increase the removal capability of facultative ponds, not only or easily biodegradable compounds but also for xenobiotics. PMID- 12894751 TI - [Clopidogrel as adjunctive medication in patients with acute coronary syndrome]. PMID- 12894752 TI - [Indications for echocardiography in ischemic cerebral insult]. PMID- 12894753 TI - Perchlorate retention and mobility in soils. AB - Adsorption and release of perchlorate in a variety of soils, minerals, and other media were studied when the solid media were exposed to low and high aqueous solutions of perchlorate salts. Low level ClO4- exposure was investigated by subjecting triplicate 5.0 g portions of a solid medium (38 different soils, minerals, or dusts) to 25 mL of an aqueous ammonium perchlorate (NH4ClO4) solution containing 670 ng mL(-1) (6.8 microM) perchlorate. This corresponds to a perchlorate-to-soil ratio of 3.4 microg g(-1) (34 nmol g(-1)). At this level of exposure, more than 90% of the perchlorate was recovered in the aqueous phase, as determined by ion chromatography. In some cases, more than 99% of the perchlorate remained in the aqueous phase. In some cases, the apparent loss of aqueous perchlorate was not clearly distinguishable from the variation due to experimental error. The forced perchlorate anion exchange capacities (PAECs) were studied by soaking triplicate 5.0 g portions of the solid media in 250 mL of 0.20 M sodium perchlorate (NaClO4) followed by repeated deionized water rinses (overnight soaks with mixing) until perchlorate concentrations fell below 20 ng mL(-1) in the rinse solutions. The dried residua were leached with 15.0 mL of 0.10 M sodium hydroxide. The leachates were analyzed by ion chromatography and the perchlorate concentrations thus found were subsequently used to calculate the PAECs. The measurable PAECs of the insoluble and settleable residua ranged from 4 to 150 nmol g(-1) (micromol kg(-1)), with most in the 20-50 nmol g(-1) range. In some soils or minerals, no sorption was detectable. The mineral bentonite was problematic, however. Overall, the findings support the widely accepted idea that perchlorate does not appreciably sorb to soils and that its mobility and fate are largely influenced by hydrologic and biologic factors. They also generally support the idea that intrasoil perchlorate content is depositional rather than sorptive. On the other hand, sorption (anion replacement) of perchlorate appears to occur in some soils. Therefore, the measurement of perchlorate in soils requires accounting for ion exchange phenomena; leaching with water alone may give inaccurate results. If perchlorate anion exchange is confirmed to be negligible, then leaching procedures may be simplified accordingly. PMID- 12894754 TI - Coencapsulation of hepatocytes and bone marrow stem cells: in vitro conversion of ammonia and in vivo lowering of bilirubin in hyperbilirubemia Gunn rats. AB - BACKGROUNDS/AIMS: This study investigates the ammonia removal capacity of coencapsulated hepatocytes and bone marrow stem cells in culture, and the treatment effect on hyperbilirubinemia Gunn rats when transplanted. METHODS: The hepatocytes and bone marrow stem cells isolated from Wistar rats were encapsulated alone or coencapsulated. In vitro, the encapsulated cells were cultured in media supplemented with 2.4 mMol/L concentration of ammonium chloride and the ammonia removal and urea synthesis were evaluated. In vivo, the encapsulated cells were transplanted intraperitoneally into hyperbilirubinemia Gunn rats and plasma bilirubin levels were measured before and after transplantation at intervals of 85 days. RESULTS: The ammonia removal capacity was maintained longer in the different ammonia concentration media in the coencapsulated hepatocytes and bone marrow cells culture. In the coencapsulation transplantation group, the plasma bilirubin levels were significantly lower than those in the group of hepatocytes encapsulation transplantation during the period of 3 to 10 weeks posttransplantion. CONCLUSIONS: The coencapsulated heaptocytes and bone marrow cells when compared to encapsulated hepatocytes could improve the maintenance of hepatocyte function both in vitro of ammonia removal in culture, and in vivo of the lowering the Gunn rats blood total bilirubin when transplanted. PMID- 12894755 TI - Conceptual paper for modelling protein and lipid accretion in different body parts of growing and fattening pigs. AB - The objective of this review is to outline those parts of modelling approaches in pig production which are not highly developed: these are the partitioning of protein and lipid accretion in different anatomical body parts. The authors introduce present models with a critical evaluation and draw some conclusions for further developments. Based on present knowledge this paper demonstrates the process of protein and fat accretion in different body compartments in pigs and influencing factors. A further aim is to assist in the conceptual development of a new pig model, which is more detailed, precise and accurate than currently available models. Exsisting models are generally deficient with regard to the translation of lipid and protein gain into lean and fatty tissue. Only assumed values for this translation have been used so far and the concepts underlying these values are not well understood. Therefore, it may be appropriate to develop a compartimental model to predict protein and fat deposition in growing and fattening pigs. With this new approach the model can supply sufficiently the changing consumer demands regarding to the possibility of meat quality prediction. PMID- 12894756 TI - The effect of oil as a dietary component on in vitro dissolution of an acid labile drug. AB - The non-benzodiazepine-like anxiolytic agent deramiclane fumarate (EGIA-3886) was used to demonstrate that the presence of high oil/fat content in dissolution media serves as a barrier against accelerated drug degradation in acidic media. PMID- 12894757 TI - Does use of the calendar in surveys reduce heaping? AB - This study examines heaping of reported durations of three postpartum variables in five countries based on two methods of data collection in retrospective surveys. Two Demographic and Health Surveys were conducted for each of the countries, one that used a five-year month-by-month calendar to record the durations and a subsequent survey in which duration information was collected from simple questions in the body of the questionnaire. Heaping indexes were calculated for breastfeeding, amenorrhea, and abstinence at six, 12, 18, and 24 months postpartum. Use of the calendar was associated with a significant reduction in the proportion of heaped responses for both breastfeeding and amenorrhea, and the effect increased for longer durations. To obtain more accurate information about these variables, the authors recommend the use of the calendar. PMID- 12894758 TI - Life before birth: effects of cortisol on future cardiovascular and metabolic function. AB - The concept of fetal programming is an area that is now under rigorous investigation in many laboratories throughout the world. We need to engender a fascination in all segments of society, not just pregnant women, about life in the womb. CONCLUSION: Everyone needs to understand that improving the condition of the fetus will have personal, social and economic benefits. The time has come to realize that, in a sense, it is not just women who are pregnant but it is the family and the whole of society. PMID- 12894759 TI - [Advances in the study of babu patch--a kind of traditional Chinese medical adhesive plasters]. PMID- 12894760 TI - [Promoting the development of ophthalmology by introducing advanced biotechnology]. PMID- 12894761 TI - [Basic techniques and environment of cell culture in ophthalmology]. PMID- 12894762 TI - [Biological effects of docosahexaenoic acid on the retina]. PMID- 12894763 TI - Pediatric anesthesiology. PMID- 12894764 TI - Endovenous laser treatment. PMID- 12894765 TI - Cloned Shiga toxin 2 B subunit induces apoptosis in Ramos Burkitt's lymphoma B cells. PMID- 12894766 TI - FDNY furor. PMID- 12894767 TI - FDNY furor. PMID- 12894768 TI - [Severe acute respiratory syndrome (SARS)]. PMID- 12894769 TI - Chaulmoogra oil and the treatment of leprosy. PMID- 12894770 TI - Nisei pharmacists in World War II. PMID- 12894771 TI - Evidence from the Derby Mercury, 1790-1800 for dental services in the North Midlands. PMID- 12894772 TI - Standard operating procedures in community pharmacy: an historical example. PMID- 12894773 TI - Some notes on an early nineteenth century manuscript medical receipt book. AB - There are 97 remedies listed, including 11 veterinary ones. These numbers include several that are duplicates. The commonest types of medicament are salves or ointments, of which there are ten, but these ten do not include ointments for specific complaints such as haemorrhoids or scurvy. The most frequently found cures are for the itch (10), rheumatism (5), gravel (4), pain (4), and piles (3), all the others having only one or two entries. They were intended to treat 39 human complaints and 9 animal ones. In addition there were formulae for killing lice, making rat poison, and preparing damson wine! The number of different medicaments that were used in the recipes was relatively small, but more than were to be found in the smaller sizes of domestic medicine cabinet. In 1820 Reece's Traveller's Dispensary that was flat and would fit in the pocket of a carriage, only contained ten drugs plus court plaster, lint, scales and weights with a book of directions and cost L3.10s.0d. (L3.50). The Lady's Dispensary which contained twenty medicines, including two pills, with some dispensing equipment and a book of directions cost L5.10s.0d. (L5.50). In all, he listed twenty different cabinets and a sea medicine chest ranging in price from L3.10s.0d. to L32.10s.0d. They included ones suitable for the family, country clergymen, and travellers on the continent and in the tropics. In 1862 Savory and Moore stocked a range of sixty-seven different medicine chests and cases in rosewood, mahogany, walnut, boxwood and leather that were fitted with 'modern appliances and conveniences adapted for the requirements of families, clergymen, officers, owners of yachts, and travellers.' Unfortunately no prices are quoted. I think that we can safely assume that the treatment received at the hands of Evan Jones was likely to be rather rough and ready when compared to the ministrations of a physician, surgeon, clergyman or local 'Lady Bountiful', but, nevertheless, must have been of great value to those who could not afford professional treatment. PMID- 12894774 TI - Horse balls and lethal chambers: the veterinary chemist in Great Britain 1900 to 1948. PMID- 12894775 TI - [History and uses of honey, mead and hive products]. AB - From Antiquity to marketing of cane sugar, and beet sugar in particular, honey was greatly consumed for its sweetening power; it was also employed for its therapeutic properties. Actually, it is always used in feeding and dietetic, and only as popular remedy. The hive products: pollen, propolis, royal jelly, introduced for few decades in dietetic meet show a great commercial passion; the beeswax was excipient in beauty care. These different aspects are developed in this study. PMID- 12894776 TI - [Honey's virtues in theriacs]. AB - Electuaries and theriacs include an important proportion of honey in their ingredients This one has a double purpose. It is an exicipient and a curative agent. On the other hand it is used as preservative, blinder and sweetener and on the other hand it is recommended because of its numerous healing virtues. Physicians take care of the selection and preparation's conditions to have an optimal quality of the honey and they consider its real nature got during its elaboration. Honey will be a perfect food and drug if it was not subjected to the digestion's action after its absorption. PMID- 12894777 TI - [Low density lipoprotein apheresis therapy during pregnancy]. PMID- 12894778 TI - [Muscular toxicity of antilipemic agents revealed during the course of hypothyroidism]. PMID- 12894779 TI - [Enhance the development of hematology to welcome new century]. PMID- 12894780 TI - [To further improve the treatment effect of acute leukemia]. PMID- 12894781 TI - The prospect of human cloning: improving nature or dooming the species? PMID- 12894782 TI - [To lay stress on the clinical application of thrombosis and hemostasis examination]. PMID- 12894783 TI - Disability, sexuality and access to services. PMID- 12894784 TI - "Disabled people aren't interested in sex...". PMID- 12894785 TI - [The use of honey in the simple and composed drugs at Rhazes]. AB - Razi or Rhazes for Latin people is one of the great scientists of ninth century. He left major works in medicine, pharmacy and in alchemy. We examine here, the use of honey by this scientist as a simple drug and as one of the essential substances included in composed medicines. That leads us to examine the notion of simple and of composed in this physician and alchemist. PMID- 12894786 TI - [Myelodysplastic syndromes and preleukemia]. PMID- 12894787 TI - [Studies on glucose-6-phosphate dehydrogenase (G6PD) deficiency in China: forty years retrospection and perspective]. PMID- 12894788 TI - [Bee, wax and honey]. AB - The archives of Nantes contain two manuscripts of the XVIIth century from which we found 63 formulae which enter bees, honey and wax. Our study concerns these various galeniques forms for internal use or external used in therapeutics and in beauty care. PMID- 12894789 TI - Medicolegal implications of board certification. PMID- 12894790 TI - [The plan of Mesaize's chemistry course]. AB - Pierre-Francois Mesaize was an apothecary, and later a pharmacist, who redacted the plan of Chemistry lessons he delivered from 1777 to 1781, in Rouen. This course was divided in three parts corresponding to the three kingdoms of nature. The items presented in this plan were compared to those which were treated in a Chemistry book published by Antoine Baume in 1773, for the mineral kingdom, and to the famous Elements de pharmacie of the same author for vegetable and animal kingdoms. The themes were almost the same except for foods chemistry and for biochemistry in which Mesaize was more interested than Baume. PMID- 12894791 TI - Regarding "Optimal therapy for advanced chronic venous insufficiency". PMID- 12894792 TI - Regarding "The safety, efficacy and pharmacoeconomics of low-dose alteplase compared with urokinase for catheter-directed thrombolysis of arterial and venous occlusions". PMID- 12894793 TI - Disseminated intravascular coagulation. PMID- 12894794 TI - [Pharmacist and chemist Henri Braconnot]. AB - Henri Braconnot was born in Commercy in 1780. When he was 13 years old, he was bound apprentice in the pharmacy of Graux in Nancy. He remained here for two years, learning pharmacy, chemistry and botany. When the army called him, his position in Strasbourg allowed him to study. After it, he came to Paris and learned in some schools, particularly the Ecole gratuite de pharmacie. Returned in Nancy, forever, in 1802, he was then named as the chairman of the botanical garden and a member of the academy of the town, and he began his results in plant chemistry. He worked almost until his death that occurred in January 1855. Braconnot's work is essentially devoted to chemistry. His name is attached to the extractive principles of vegetables. Some orientations of his research are: plant acids, fats, extractive compounds that may have conduct him to alkaloids, sugar obtained from saw-dust and rages, other organic plant chemicals, nitration of cellulose, and some others such as colours chemistry, hydrology, physiological chemistry, toxicology, mineralogy. He was never received as an apothecary, but pharmacy has always considered that he belonged to it. PMID- 12894795 TI - [Childhood's anemia]. PMID- 12894796 TI - More need for the "old" in the public health research in the Americas. PMID- 12894797 TI - FDA moves to curb drug counterfeiting. PMID- 12894798 TI - Conference deplores corporate influence on academic science. Speakers argue that corporate funds should be separated from science to prevent undue influence. PMID- 12894799 TI - Japan steps up fight against tobacco industry. Antismoking lobby fights for public's hearts, lungs, and minds. PMID- 12894800 TI - [Apothecaries and communal saltpeter works: the example of Joachim Proust (1751 1819)]. AB - In 1793, our country was besieged by almost the entire Europ. During the year following December 4, 1793, 6 000 private and communal saltpeter works had produced 16 764 039 lb of saltpeter. They had been demanded by the Committee of Public Safety for supplying the gunpowder factories. Their management was generally given to apothecaries. Joachim Proust, who was the brother of Joseph Louis Proust (definite proportions rule), was appointed to the district of Angers. From 1778 to 1788 he had worked in Paris under Hilaire Marin Rouelle and met the greatest chemists. The district of Angers confronted the civil war of Vendee. Joachim Proust chose to favour professional saltpeter makers. The saltpeter collect, which has reached the smallest villages, has been a significant factor of national unity around the revolutionary ideas. PMID- 12894801 TI - [The Boulducs' dynasty, apothecaries in Paris in XVIIth and XVIIIth centuries: Jean Francois Boulduc (February 20, 1728 - August 18, 1769)]. AB - Jean-Francois Boulduc was Gilles-Francois Boulduc's illegitimate child. At his death his property inventory revealed numerous documents (baptism certificate, ill, etc.) that allowed us to specify his professional activity. He practised galenic and chemical pharmacy trade rue des Boucheries in Paris and went into partnership with two apothecaries Claude Pia and Michel Depene successively. First apothecary of King Louis XV, he provided the Court in Versailles and Marley with medicines that he prepared in his pharmacy in Paris or in a laboratory in his house in Marly-le-Roi. He died in Paris on August 18, 1769 without progeny. PMID- 12894802 TI - [The International Academy of the History of Pharmacy 1952-2002]. PMID- 12894803 TI - Notification to a prior publication. PMID- 12894805 TI - Bibliography. Current World Literature. Occupational Disease. PMID- 12894804 TI - Columbia: point of departure. PMID- 12894806 TI - Bibliography. Current world literature. Pediatric asthma and development of atopy. PMID- 12894807 TI - Source memory enhancement for emotional words. AB - The influence of emotional stimuli on source memory was investigated by using emotionally valenced words. The words were colored blue or yellow (Experiment 1) or surrounded by a blue or yellow frame (Experiment 2). Participants were asked to associate the words with the colors. In both experiments, emotionally valenced words elicited enhanced free recall compared with nonvalenced words; however, recognition memory was not affected. Source memory for the associated color was also enhanced for emotional words, suggesting that even memory for contextual information is benefited by emotional stimuli. This effect was not due to the ease of semantic clustering of emotional words because semantically related words were not associated with enhanced source memory, despite enhanced recall (Experiment 3). It is suggested that enhancement resulted from facilitated arousal or attention, which may act to increase organization processes important for source memory. PMID- 12894808 TI - The informational impact of mood on effort mobilization: a study of cardiovascular and electrodermal responses. AB - The impact of mood on effort quantified as autonomic adjustments was investigated in an experiment. The authors induced positive versus negative moods with either 1 of 2 mood induction procedures (music vs. autobiographical recollection) that differed in the extent of required effort. Then participants performed an achievement task after demand appraisals were made. Results were as predicted. During the mood inductions, autonomic reactivity (systolic blood pressure [SBP], diastolic blood pressure, heart rate, skin conductance responses) was stronger in the relatively effortful recollection conditions than in the relatively effortless music conditions. Mood valence had no impact here. But in the context of task performance, the authors found (a) mood congruency effects on the demand appraisals that reflected subjectively higher demand in a negative than in a positive mood, and (b) stronger SBP reactivity in a negative mood compared with a positive mood. Furthermore, SBP reactivity during task performance was correlated with achievement. PMID- 12894809 TI - Rumination as a common mechanism relating depressive risk factors to depression. AB - Rumination was examined as a potential common mechanism linking risk factors with depression. Initially nondepressed individuals (N = 137) were assessed for presence of a ruminative response style and 4 other hypothesized risk factors for depression. They were followed for 2.5 years. Negative cognitive styles, self criticism, dependency, neediness, and history of past depression were all significantly associated with rumination. Rumination mediated the predictive relationships of all risk factors except dependency with the number of prospective Diagnostic and Statistical Manual of Mental Disorders (3rd ed., rev.; American Psychiatric Association, 1987) major depressive episodes (MDEs; definite and subthreshold) during the follow-up period. In contrast, private self consciousness did not mediate any relationships between risk factors and subsequent MDEs. Thus, rumination, as a special kind of self-focus, may act as a general proximal mechanism through which other vulnerability factors affect depression. PMID- 12894810 TI - Emotional and personality changes following cingulotomy. AB - The effects of bilateral anterior cingulate cortex (ACC) lesions on emotional and personality functioning were studied. Patients undergoing cingulotomy for chronic intractable pain were assessed on the Minnesota Multiphasic Personality Inventory (MMPI), the Profile of Mood States (POMS), cognitive tests, and pain ratings, pre and postsurgically. Patients with intractable pain, not treated with cingulotomy, served as controls. Cingulotomy patients experienced reductions in POMS-Tension, POMS-Anger, and MMPI Scale 7 (Psychasthenia) compared with baseline and the controls. POMS-Tension was significantly correlated with attention intention performance. The results indicate that the ACC modulates emotional experience, related to self-perceived tension, and that there is relationship between the emotional and the attentional effects of cingulotomy. PMID- 12894811 TI - Social context effects on facial activity in a negative emotional setting. AB - Evidence for A. J. Fridlund's (e.g.. 1994) "behavioral ecology view" of human facial expression comes primarily from studies of smiling in response to positive emotional stimuli. Smiling may be a special case because it clearly can, and often does serve merely communicative functions. The present experiment was designated (a) to assess the generalizability of social context effects to facial expressions in response to negative emotional stimuli and (b) to examine whether these effects are mediated by social motives, as suggested by the behavioral ecology view. Pairs of friends or strangers viewed film clips that elicited different degrees of sad affect, in either the same or a different room; a control group participated alone. Dependent variables included facial activity, subjective emotion, and social motives. Displays of sadness were influenced by stimulus intensity and were lower in all social conditions than in the alone condition. Unexpectedly, social context effects were also found for smiling. PMID- 12894812 TI - A functional MRI study of human amygdala responses to facial expressions of fear versus anger. AB - Functional magnetic resonance imaging (fMRI) of the human brain was used to compare changes in amygdala activity associated with viewing facial expressions of fear and anger. Pictures of human faces bearing expressions of fear or anger, as well as faces with neutral expressions, were presented to 8 healthy participants. The blood oxygen-level dependent (BOLD) fMRI signal within the dorsal amygdala was significantly greater to Fear versus Anger, in a direct contrast. Significant BOLD signal changes in the ventral amygdala were observed in contrasts of Fear versus Neutral expressions and, in a more spatially circumscribed region, to Anger versus Neutral expressions. Thus, activity in the amygdala is greater to fearful facial expressions when contrasted with either neutral or angry faces. Furthermore, directly contrasting fear with angry faces highlighted involvement of the dorsal amygdaloid region. PMID- 12894813 TI - Perceptual asymmetries reflect developmental changes in the neuropsychological mechanisms of emotion recognition. AB - To study how perceptual asymmetries in the recognition of emotion reflect developmental changes in processing affective information, a fused rhyming dichotic word test with positive, negative, and neutral stimuli was administered to adults and children. Results suggested that the hemisphere in which affective information is initially processed affects the strength of perceptual asymmetry and that children's perceptual processing of emotional information is constrained by limited computational resources. Another experiment ruled out effects of volitional shifting of attention to emotional stimuli. These data further confirm that emotional processing involves integration of neural systems across brain regions, including distributed systems that support arousal and recognition. General developmental factors, such as processing capacity, contribute to the coordination of multiple systems responsible for processing emotional information. PMID- 12894814 TI - Electrochemical tagging of urate: developing new redox probes. AB - The electro-oxidation of uric acid in the presence of nucleophilic species is shown to produce conjugates that can aid our understanding of antioxidant interactions and also provide opportunities for advancing electroanalytical detection strategies involving purine species. PMID- 12894815 TI - A metabonomic investigation of hepatotoxicity using diffusion-edited 1H NMR spectroscopy of blood serum. AB - It has been shown extensively, that chemometric investigations of 1H NMR spectra of rat urine taken from animals dosed with model toxins produce characteristic patterns of metabolic responses and that this permits the identification of biomarkers of toxic response and regeneration. To date, metabonomic methods have been mainly optimised for urine which contains mainly low molecular weight moieties, and thus a conventional 1-dimensional 1H NMR pulse sequence is an efficient means of obtaining information-rich data. In the case of biofluids such as blood plasma or serum, which contain a wide range of macromolecules the resonances of which can overlap with peaks from small molecule metabolites, the information giving rise to sample classification can be concealed in a conventional NMR spectrum andthis presents a different analytical challenge in terms of chemometric analysis of spectral profiles. Here, the use of other types of NMR data have been investigated and it is shown that by using spectra where the peak intensities are edited according to their molecular diffusion coefficients, it is possible to improve differentiation of control animals and those treated with the model hepatotoxin, alpha-naphthylisothiocyanate (ANIT). By using diffusion-edited spectroscopy, plasma lipid moieties are less attenuated than those from small endogenous metabolites and thus the toxin-induced changes to the lipoprotein profiles are more easily detectable. PMID- 12894816 TI - Metabonomic analysis of mouse urine by liquid-chromatography-time of flight mass spectrometry (LC-TOFMS): detection of strain, diurnal and gender differences. AB - The application of HPLC-MS combined with principal components analysis (PCA) to the metabonomic analysis of mouse urine is demonstrated. Urine samples from three strains of mouse were analysed by gradient HPLC-MS combined with positive and negative electrospray time-of-flight mass spectrometry. Analysis of the resulting data using PCA enabled the samples to be discriminated between on the basis of gender, strain and diurnal variation. These preliminary results suggest that HPLC MS-based approaches may have a useful role in metabonomic analysis that complements existing approaches. PMID- 12894817 TI - Enzyme immunoassays for the investigation of protein nitration by air pollutants. AB - Two enzyme immunoassays have been developed, characterised, and applied to investigate protein nitration in birch pollen extract (BPE) and bovine serum albumin (BSA) samples exposed to air pollutants. The monoclonal antibody CAY 189542 against nitrotyrosine (raised against peroxynitrite-treated keyhole limpet hemocyanine) was characterised in an indirect competitive assay (affinity and cross-reactivities) and applied in a new one-sided enzyme immunoassay for nitrated proteins. The one-sided assay was calibrated against a nitrated BSA standard with an average of 14 nitrotyrosine residues per molecule (nitro-(14) BSA; detection limit 8.3 pmol L(-1)), and the sensitivity of the test was found to be significantly enhanced by a multivalent binding mode of the monoclonal antibody (bonus effect of multivalency). The same antibody and a polyclonal antibody against Bet v 1, the most prominent birch pollen allergen, were used in a new sandwich immunoassay for specific determination of nitrated Bet v 1. This assay was calibrated against a nitrated Bet v 1 standard with an average of 3 nitrotyrosine residues per molecule (nitro-(3)-Bet v 1; detection limit 0.2 nmol L(-1)). Bet v 1 and BSA exposed to polluted urban outdoor air and to synthetic gas mixtures containing NO2 and O3 at atmospherically relevant concentration levels were found to be efficiently nitrated within hours to days. Pronounced correlations of nitro-(14)-BSA equivalent concentrations with exposure time and with nitro-(3)-Bet v 1 equivalent concentrations in nitrated BPE samples were observed. Test experiments indicated that the efficiency of protein nitration was strongly enhanced by reactive species formed upon interaction of NO2 with O3 and H2O (e.g. NO3 and HNO3). Potential implications of protein nitration by air pollutants are outlined and discussed. PMID- 12894818 TI - Pantothenic acid quantification by a stable isotope dilution assay based on liquid chromatography-tandem mass spectrometry. AB - A stable isotope dilution assay for the quantification of free and total pantothenic acid has been developed by using [13C3,15N]-pantothenic acid as the internal standard. The three-dimensional specificity of liquid chromatography tandem mass spectrometry enabled unequivocal determination of the vitamin. Due to the very simple extraction and clean-up procedure, free pantothenic acid could be analysed within 2 h, which is much faster than by microbiological or gas chromatographic assays. For quantification of total pantothenic acid, the vitamin was liberated from its conjugates by an overnight incubation with pigeon liver pantetheinase and alkaline phosphatase. In analyses of corn flour, the intra assay coefficient of variation was 8.5% (n = 5) and 15.3% (n = 4) for free and total pantothenic acid, respectively. When pantothenic acid was added to corn starch at a level of 6 mg kg(-1), a recovery of 97.5% was found. Application of the stable isotope dilution assay to whole egg powder, hazel nuts and corn revealed similar data compared to those listed in nutrition data bases, whereas the content in mushrooms and porcine liver determined by the newly developed assay appeared to be lower and that of cocoa higher than reported in the literature. PMID- 12894819 TI - Simultaneous determination of the tobacco smoke uptake parameters nicotine, cotinine and thiocyanate in urine, saliva and hair, using gas chromatography-mass spectrometry for characterisation of smoking status of recently exposed subjects. AB - A method using gas chromatography (GC)-mass spectrometry (MS) for the simultaneous determination of the smoke uptake parameters thiocyanate, nicotine and cotinine in human tissues is reported. Nicotine, cotinine and thiocyanate, in combination with a phase-transfer catalyst, were extracted from urine, saliva and hair into dichloromethane (DCM). Thiocyanate was alkylated in the DCM-layer to form a pentafluorobenzyl derivative. The biochemical markers in DCM were directly injected into the GC system and separated on a DB-1MS column using a 9.4 min temperature program. The method was validated in urine and saliva between the limits of quantitation (1.0-15 microg ml(-1) thiocyanate, 0.010-3.0 microg ml(-1) nicotine and cotinine in urine, 0.010-1.0 microg ml(-1) nicotine and cotinine in saliva). The calibration curves were found to be linear (r > 0.996), the within- and between-day accuracy's were 83-120%, the repeatability coefficients of variation were 3-20% and the limits of detection were 0.060 ng ml(-1) thiocyanate and 0.60 ng ml(-1) nicotine and cotinine. The results of the analysis of the biomarkers in the urine of 44 volunteers were used to develop a predictive model for smoking status, using discriminant analysis. The classification model correctly classified 93.2% of cross-validated grouped cases. Saliva samples were used to confirm the results of the classification method. PMID- 12894823 TI - Identification and quantification of urinary benzo[a]pyrene and its metabolites from asphalt fume exposed mice by microflow LC coupled to hybrid quadrupole time of-flight mass spectrometry. AB - Prolonged, extensive exposure to asphalt fume has been associated with several adverse health effects. Inhaled polycyclic aromatic hydrocarbons (PAHs) from asphalt fume exposure have been suspected of inducing such effects. In this study, a bioanalytical method was proposed and evaluated to identify and quantify benzo[a]pyrene and its hydroxy-metabolites. This method is based on coupling a microflow liquid chromatography (LC) to a hybrid quadrupole orthogonal acceleration time-of-flight mass spectrometry (Q-TOFMS). In the experiment, thirty-two B6C3FI mice were exposed to asphalt fume in a whole body inhalation chamber for 10 days (4 h day(-1)) and twelve other mice were used as controls. The asphalt fume was generated at 180 degrees C and the concentrations in the animal exposure chamber ranged 175-182 mg m(-3). Benzo[a]pyrene and its metabolites of 3-hydroxybenzo[a]pyrene, benzo[a]pyrene-7,8-dihydrodiol(+/-), benzo[a]pyrene-7,8-dihydrodiol-9,10-epoxide(+/-), and benzo[a]pyrene-7,8,9,10 tetrahydrotetrol(+/-) in the urine of asphalt fume exposed mice were identified and found at 3.18 ng 100 mL(-1), 31.36 ng 100 mL(-1), 11.56 ng 100 mL(-1), 54.92 ng 100 mL(-1), and 45.23 ng 100 mL(-1) respectively. The results revealed that the urinary benzo[a]pyrene and its hydroxy-metabolites from exposed mice were at significantly higher levels (p < 0.001) than those from the control groups. Compared with several other technologies such as HPLC-UV and HPLC-fluorescence, the new method is more sensitive and selective, and it can also provide additional useful information on the structures of the metabolites. Hence, this method can be used to perform the assessment and to study the mechanisms of the adverse health effects. The fragmentation patterns established in this study can also be used to identify and quantify PAH metabolites in other biological fluids. PMID- 12894824 TI - Quantitative analysis of oxytetracycline and its 4-epimer in calf tissues by high performance liquid chromatography combined with positive electrospray ionization mass spectrometry. AB - Tetracycline antibiotics are commonly used in veterinary medicine because of their broad spectrum activity and cost effectiveness. Oxytetracycline (OTC) is one of the most important members of this antibiotic family. The purpose of this study was to develop and validate a method to determine OTC residues in edible tissues of calf. Extraction of OTC and its 4-epimer (4-epiOTC), in the presence of the internal standard demethylchlortetracycline (DMCTC), was performed using a liquid extraction with sodium succinate solution (pH 4.0), followed by protein removal with trichloroacetic acid and paper filtration. Further solid-phase extraction clean-up on an HLB polymeric reversed phase column was performed to obtain an extract suitable for LC-MS-MS analysis. Chromatographic separation of the internal standard, and especially OTC and its 4-epimer, was achieved on a PLRP-S polymeric reversed phase column, using a mixture of 0.001 M of oxalic acid, 0.5% (v/v) of formic acid and 3% (v/v) of tetrahydrofuran in water (mobile phase A) and tetrahydrofuran (mobile phase B) as the mobile phase, and at a column temperature of 60 degrees C. OTC and its 4-epimer could be identified using the MS-MS detection technique, and were subsequently quantified. The method has been validated according to the requirements of the EC at the MRL (maximum residue limit, 100 ng g(-1) for muscle, 300 ng g(-1) for liver and600 ng g(-1) for kidney), half the MRL and double the MRL levels, as well for OTC as for 4 epiOTC. Calibration graphs were prepared for all tissues and good linearity was achieved over the concentration ranges tested (r > 0.99 and goodness of fit < 10%). Limits of quantification of half the MRLs were obtained for the analysis of OTC and 4-epiOTC in muscle, liver and kidney tissues of calf. Limits of detection ranged for both components between 0.8 and 48.2 ng g(-1). The within-day and between-day precisions, expressed as RSD values, were all below the maximum allowed RSD values calculated according to the Horwitz equation. The results for accuracy fell within the -20% to +10% range. Recoveries were between 47 and 56% for OTC, and between 52 and 62% for 4-epiOTC, depending on the tissue. The method has been successfully used for the quantitative determination of OTC and 4-epiOTC in tissue samples of calves medicated with OTC by intramuscular injection. PMID- 12894830 TI - Carbon-nanotube-modified glassy carbon electrodes for amplified label-free electrochemical detection of DNA hybridization. AB - The preparation and attractive performance of carbon-nanotube modified glassy carbon (CNT/GC) electrodes for improved detection of purines, nucleic acids, and DNA hybridization are described. The surface-confined multiwall carbon-nanotube (MWCNT) facilitates the adsorptive accumulation of the guanine nucleobase and greatly enhances its oxidation signal. The advantages of CNT/GC electrodes are illustrated from comparison to the common unmodified glassy carbon, carbon paste and graphite pencil electrodes. The dramatic amplification of the guanine signal has been combined with a label-free electrical detection of DNA hybridization. Factors influencing the enhancement of the guanine signal are assessed and optimized. The performance characteristics of the amplified label-free electrochemical detection of DNA hybridization are reported in connection to measurements of nucleic-acid segments related to the breast-cancer BRCA1 gene. PMID- 12894831 TI - Amplifying the electrical hybridization signals of DNA array by multilayer assembly of Au nanoparticle probes. AB - This paper describes a versatile method for amplifying the signals of Au nanoparticle-based DNA hybridization detecting systems. The Au nanoparticles usually serve as labels to enhance DNA hybridization signal. We further assembled several layers of nanoparticles to selectively increase the number of labelled nanoparticles. Through silver enhancement, the multilayer nanoparticles may produce significantly higher amounts of metal silver on the their surfaces than the monolayer nanoparticles did. This finally accounts for the greatly enhanced DNA hybridization signal. Particularly, the amplification of electrical detection system was demonstrated here. Electrical measuring results indicated that the current values were enhanced by approximately 3 orders of magnitude, and the single nucleotide mismatch discrimination ratio was enlarged to approximately 10(9):1. PMID- 12894832 TI - Electrochemical oxidation of underivatized-nucleic acids at highly boron-doped diamond electrodes. AB - Boron-doped diamond (BDD) electrodes have been examined for the electrochemical oxidation of underivatized-nucleic acids in terms of single stranded and double stranded DNA. Cyclic voltammetry and square wave voltammetry have been used to study the oxidation reactions and to detect DNA without derivatization or hydrolysis steps. At the diamond electrode, at least two well-defined voltammetric peaks were observed for both single stranded and double stranded DNA. Diamond electrode is the first material to show a well-defined voltammetric peaks for adenine group oxidation directly in the helix structure of nucleic acid due to its wide potential window. For single stranded DNA, a third peak, related to the pyrimidine group oxidation was also observed. As-deposited diamond film with predominantly hydrogen-terminated surface exhibited superior performance over oxygen-terminated diamond in terms of sensitivity. However, by optimizing the ionic strength, sensitivity of O-terminated films could be improved. Linear calibration results have shown linearity of current with concentration in the range 0.1-8 microg mL(-1) for both guanine and adenine residues at as-deposited BDD. Detection limits (S/N = 3) of 3.7 and 10 ng mL(-1) for adenine and guanine residue in single stranded DNA, respectively, and 5.2 and 10 ng mL(-1) for adenine and guanine residue in double stranded DNA, respectively, were observed. This work shows the promising use of diamond as an electrochemical detector for direct detection of nucleic acids. The results also show the possibility of using the oxidation peak current of adenine group that is more sensitive for the direct detection of nucleicacids. PMID- 12894833 TI - Studies on the electrochemical behavior of cytochrome c and its interaction with DNA at a Co/GC ion implantation modified electrode. AB - The electrochemical behavior of cytochrome c (cyt c) and its interaction with DNA at a Co/glassy carbon (GC) ion implantation modified electrode were studied by linear sweep and cyclic voltammetry. In 0.005 mol dm(-3) Tris-0.05 mol dm(-3) NaCl buffer solution (pH = 7.10), a sensitive reduction derivative peak of cyt c was obtained by linear sweep voltammetry. The peak potential was 0.032 V (SCE). The peak current was proportional to the concentration of cyt c. The electrode process was quasi-reversible with adsorption. The electrode reaction rate constant k and the electron transfer coefficient a of cyt c were 4.42 s(-1) and 0.47, respectively. AES and XPS experiments showed that Co was implanted into the surface of the GC electrode (GCE). The implanted Co formed Co-C, which catalyzed the reduction of cyt c. The reaction of DNA with cyt c led to an electrochemically active complex, which resulted in an increase in the reduction current of cyt c. After adding DNA into the solution containing cyt c, the electrode process was still quasi-reversible with adsorption. PMID- 12894836 TI - Nondestructive discrimination of ivories and prediction of their specific gravity by Fourier-transform Raman spectroscopy and chemometrics. AB - Fourier-transform (FF) Raman spectroscopy and chemometrics were used for nondestructive analysis of ivories. The discrimination of five kinds of ivories, two subspecies of African elephant, mammoth, hippopotamus, and sperm whale, was investigated, and a calibration model for predicting their specific gravity was developed. FT-Raman spectra were measured in situ for them and chemometrics analyses were carried out for the 3050-350 cm(-1) region. The five kinds of ivories were clearly discriminated from each other on the scores plots of two or three principal components (PCs) obtained by principal component analysis (PCA). The loadings plot for PC 1 shows that the discrimination relies on the content ratio of organic collagenous protein and inorganic hydroxyapatite of ivories. The loadings plot for PC 2 shows that bands due to the CH3 and CH2 stretching modes of the protein also play a role in the discrimination. Using partial least squares regression (PLSR), we developed a calibration model that predicts the specific gravity of the ivories from the FT-Raman spectra. The correlation coefficient and root mean square error of cross validation (RMSECV) of this model were 0.980 and 0.024, respectively. PMID- 12894838 TI - Near-infrared spectroscopy for liquids of microliter volume using capillaries with wall transmission. AB - In the present study we propose a capillary tube method for measuring near infrared (NIR) spectra of microliter liquid and solution samples. This method enables one to measure NIR spectra of liquids and solutions of only 2.5 microl. As an example of the capillary tube method, Fourier-transform NIR spectra of benzene were measured using a capillary tube with a diameter of 1 mm. Positions and intensities of the NIR bands in the spectra obtained by the capillary tube were almost identical to those measured using a quartz cuvette cell. Moreover, capillary NIR spectra of human blood serum obtained from a patient with backbone neoplasm were also measured. Subtraction of the NIR spectrum of water from those of human blood serum develops unambiguously some NIR bands due to blood components such as proteins and glucose. Our results suggest that capillary NIR spectroscopy will open new areas of NIR applications for small amounts of liquid and solution samples. PMID- 12894840 TI - Total luminescence spectroscopy with pattern recognition for classification of edible oils. AB - Total luminescence spectroscopy combined with pattern recognition has been used to discriminate between four different types of edible oils, extra virgin olive (EVO), non-virgin olive (NVO), sunflower (SF) and rapeseed (RS) oils. Simplified fuzzy adaptive resonance theory mapping (SFAM), traditional back propagation (BP) and radial basis function (RBF) neural networks provided 100% classification for 120 samples, SFAM was found to be the most efficient. The investigation was extended to the adulteration of percentage v/v SF or RS in EVO at levels from 5% to 90% creating a total of 480 samples. SFAM was found to be more accurate than RBF and BP for classification of adulterant level. All misclassifications for SFAM occurred at the 5% v/v level resulting in a total of 99.375% correctly classified oil samples. The percentage of adulteration may be described by either RBF network (2.435% RMSE) or a simple Euclidean distance relationship of the principal component analysis (PCA) scores (2.977% RMSE) for v/v RS in EVO adulteration. PMID- 12894841 TI - Study on Schiff base complexes-cellular DNA interactions by a novel system of Hadamard transform fluorescence image microscopy. AB - A novel system of Hadamard transform microscopic fluorescence imaging for single cells is presented, based on which the DNA ploidy of rat hepatocyte was quantitatively measured. The result shows that diploid rat hepatocyte has a stable DNA content, thus diploid rat hepatocyte was used to investigate the binding of five clinical anticancer agents, vincristine, cyclophosphamide, nitrogen mustard, cis-diamminedichloroplatinum(n) (CDDP) and mitomycin-C, with cellular DNA when acridine orange (AO) was used as the competitive fluorescence probe. Based on this model, some Schiff base complexes-cellular DNA interactions were investigated. The results indicate that all the twenty-two compounds, including Schiff base ligands of N-2-hydroxy-naphthaldehyde with D-glucoamine (NG) and the complexes of 3d-transitional metals ions with NO and with D glucoamine (Glu) and the mixed complexes of NG and Glu series with alpha-glycine (GNG), have the ability to enter the cell membrane and interact with cellular DNA. Four of the compounds, CuGlu, Fe(II)NG, Fe(III)NG and CuGluG can intercalate with DNA like AO does and depress AO-DNA fluorescence to 70% or lower. An in intro UV-visible spectroscopic study on the compound-DNA spectra testified the above results and suggests that diverse interaction mechanisms coexist for all these complexes except intercalating mode. This study presents a new in vitro method for initial screening of anticancer compounds. PMID- 12894842 TI - Static pressurised liquid extraction of nitrated polycyclic aromatic hydrocarbons from soils with on-line filtration-preconcentration prior to gas chromatography mass spectrometry detection. AB - Nitrated polycyclic aromatic hydrocarbons have been extracted from both spiked and natural contaminated soils by using static pressurised liquid extraction. The variables governing the extraction (namely, pressure, extraction time, temperature and number of static extraction cycles) have been optimised using the experimental design methodology. A laboratory-made pressurised liquid extractor has been used allowing its coupling to a filtration-preconcentration system through a flow injection interface, thus providing a partially automated approach. Gas chromatography with MS-MS ion preparation mode has been used for individual separation-identification/quantification providing both high selectivity (no interferences were observed) and sensitivity (detection limits of low pg). The comparison of the proposed approach with the reference EPA Method 3540 has shown that both methods provide similar efficiencies but with dramatic saving of time by the former (30 min vs. 24 h). The use of water as leaching agent avoids the use of organic solvents, thus providing an environmentally friendly method. PMID- 12894843 TI - Effective pretreatment of human serum samples for dioxin analysis by solid phase extraction and blue-chitin column cleanup. AB - For the analysis of dioxins, polychlorinated dibenzo-p-dioxins/dibenzofurans (PCDDs/Fs) and coplanar polychlorinated biphenyls (Co-PCBs), devising simple and economical methods is important, especially for mass screening of human exposure. Pretreatment of samples, namely the extraction and cleanup methods that are widely used at present, needs to be improved for savings in time, manpower, and solvent consumption. In the present study, we applied solid phase extraction (SPE) using octadecyl (C18) and a blue-chitin column in place of liquid-liquid extraction (LE) and an active-carbon column with serum samples, frequently used for assessment of human exposure. Efficacy of the new pretreatment methods was demonstrated by successful high resolution gas chromatography-high resolution mass spectrometry (HRGC-HRMS) of the major 17 PCDDs/Fs and 12 Co-PCBs that are on the list of WHO/IPCS (1997) hazardous dioxins with toxic equivalent factor (TEF) values. SPE is timesaving and requires less manpower and organic solvent as compared with the LE that is presently widely used. Concerning cleanup with blue chitin, the amount of toluene applied as eluent could be reduced to 1/3, as compared with the active-carbon case. The combination of SPE and blue-chitin for pretreatment of serum saves time and manpower, is accurate and uses less organic solvent than LE with active carbon cleanup. PMID- 12894844 TI - Development and functions of natural killer cells. AB - Over the last decade, progress in molecular and cellular biology and gene targeting techniques has removed veils from the mysteries of natural killer (NK) cell development and function. NK cells are derived from hematopoietic stem cells, for which stem cell factor or Flt3 ligand is required in the early stage of differentiation to NK cell progenitors. Interleukin 15 then plays a crucial role for differentiation and/or maturation of NK progenitors into functional NK cells. Several members of the zinc finger, ETS, and interferon regulatory factor transcription factor families are also involved in the lineage commitment of hematopoietic stem or progenitors into NK cells. Animal models as well as patients deficient in NK cells have provided formal evidence that NK cells play an important role in vivo for innate immunity against tumors and viral infections and for linkage to adaptive immunity. Moreover, recent studies have revealed novel human NK cell subsets in peripheral blood that have the phenotypical characteristics CD3- CD16+ CD56+ and CD3- CD16- CD56bright, which are mainly involved in cytotoxicity and cytokine-mediated immunoregulation, respectively. PMID- 12894845 TI - Biology and clinical impact of human natural killer cells. AB - Natural killer (NK) cells, through elaboration of cytokines and cytolytic activity, are critical to host defense against invading organisms and malignant transformation. Two subsets of human NK cells are identified according to surface CD56 expression. CD56dim cells compose the majority of NK cells and function as effectors of natural cytotoxicity and antibody-dependent cellular cytotoxicity, whereas CD56bright cells have immunomodulatory function through secretion of cytokines. For a long time, NK cells have held promise for cancer immunotherapy because, unlike T-lymphocytes, NK cells can lyse tumor cells without tumor specific antigen recognition. To date, NK cell therapy, largely focused on in vivo expansion and activation with cytokines, has met with only modest success. However, recent understanding of the importance of NK receptors (NKR) for recognition and lysis of tumor cells while normal cells are spared suggests novel therapeutic strategies. The balance of inhibitory and activating signals through surface receptors that recognize major histocompatibility complex class I and class I-like molecules on target cells determines whether NK cells activate killing. Identification of NKR ligands and their level of expression on normal and neoplastic cells has important implications for the rational design of immunotherapy strategies for cancer. We review recent development in the biology and clinical relevance of NK cells in cancer immunotherapy. PMID- 12894846 TI - Leukemia and lymphoma of natural killer lineage cells. AB - Natural killer (NK) cells are lymphocytes with a large granular lymphocyte morphology, a CD3- CD56+ phenotype, a non-major histocompatibility complex restricted cytotoxicity, and germline configuration T-cell receptor genes. NK cell lineage tumors originate from either precursor NK cells or mature NK cells. Tumors originating conceivably from precursor NK cells include myeloid/NK cell precursor acute leukemia, precursor NK cell acute lymphoblastic leukemia, and blastic NK cell lymphoma. However, because the developmental pathway of normal NK cells and the characteristics of these NK precursors are not fully understood, the definition and characterization of the tumors are only provisional. Tumors of mature NK cell origin include aggressive NK cell leukemia/lymphoma, nasal-type NK cell lymphoma, and chronic NK lymphocytosis, but the last disorder seems to be reactive in most cases. Because NK cell tumors are rare and difficult to manage, vigorous studies are required for their understanding and management. PMID- 12894847 TI - Diagnosis and treatment of Epstein-Barr virus-associated natural killer cell lymphoproliferative disease. AB - Epstein-Barr virus (EBV) exhibits tropism for both lymphocytes and epithelial cells and can induce both replicative (productive/lytic) and latent (persistent) infections that result in a variety of human diseases. With regard to lymphocytes, latent EBV infection is linked to development of heterogeneous lymphoproliferative disease (LPD), such as B-cell LPD and T-cell/natural killer cell (T/NK cell) LPD. Unlike B-cell LPD, LPD derived from T-cells and NK cells sometimes has overlapping clinical symptoms, as well as histologic and immunophenotypic features, because both types of cells are derived from a common precursor. However, determination of cell lineage is important in classification of lymphoid neoplasms, and combined modern techniques allows us to distinguish NK cell LPD from T-cell LPD in most instances. Because NK cell LPD seems to be heterogeneous in terms of clinical features, prognosis, and diagnosis and has a monoclonal or polyclonal (or oligoclonal) nature, this review attempts to clarify recent research and clinical findings and to establish diagnostic and therapeutic strategies. PMID- 12894848 TI - Treatment of acute promyelocytic leukemia and other hematologic malignancies with arsenic trioxide: review of clinical and basic studies. AB - Acute promyelocytic leukemia (APL) is now the most potentially curable subtype of acute myeloid leukemia in adults because of the introduction of novel approaches in the management of this disease. All-trans-retinoic acid (ATRA)-based therapy is now the first-choice treatment of patients presenting with de novo APL, and clinical studies have shown that nearly all patients who receive ATRA therapy achieve complete remission. However, approximately 20% to 30% of APL patients eventually have relapses with resistance to further ATRA treatment. Arsenic trioxide (As2O3 [ATO]) has been established as highly effective therapy for patients with APL, even for those with disease refractory to ATRA. Furthermore, results of recent studies have suggested a broad therapeutic potential for ATO in the treatment of hematologic malignancies beyond APL. In this review, we discuss the clinical activity and multiple mechanisms of ATO therapy in the management of APL and other hematologic neoplasms. PMID- 12894849 TI - Mutational analysis of the WASP gene in 2 Korean families with Wiskott-Aldrich syndrome. AB - Wiskott-Aldrich syndrome (WAS), an X-linked disorder characterized by thrombocytopenia with undersized platelets, eczema, and immune deficiency, is caused by mutations in the WASP gene. In this study, we investigated WASP gene mutations and WASP protein expression in 2 unrelated Korean WAS patients. Flow cytometry was used to evaluate WASP expression in lymphocytes. Two previously reported nonsense mutations (Arg211stop and Arg13stop) were identified in this study, a finding that suggested these codons are mutational hotspots. Both mothers showed normal WASP expression in flow cytometric analysis, even though they had heterozygotic patterning, which is indicative of carrier status. Furthermore, an X-chromosome inactivation assay revealed that these carrier mothers had skewed X inactivation. To our knowledge, this is the first report on molecular diagnosis of WAS in Korea. In addition, we detected normal WASP expression in lymphocytes from carrier mothers, a finding consistent with the data on skewed X inactivation. PMID- 12894850 TI - Identification of an SH2D1A mutation in a hypogammaglobulinemic male patient with a diagnosis of common variable immunodeficiency. AB - Common variable immunodeficiency (CVID) is a highly heterogeneous disease with an unpredictable pattern. CVID appears to have an immunologic and clinical phenotype similar to some hereditary humoral immunodeficiencies, including X-linked lymphoproliferative disease (XLP). The differential diagnosis of CVID and XLP is clinically of importance, because the two diseases have markedly different prognoses and treatment. The recent identification of the XLP gene, known as SH2D1A, has permitted a definitive diagnosis of XLP. In this report, we describe a male patient with XLP who initially received a diagnosis of CVID and developed a fatal course. Using genetic analysis, we confirmed that the patient harbored the SH2D1A gene mutation. The results support the notion that the possibility of a SH2D1A gene mutation should be considered in hypogammaglobulinemic male patients before a diagnosis of CVID is made. PMID- 12894851 TI - Spontaneous and rapid reexpression of functional CXCR4 by human steady-state peripheral blood CD34+ cells. AB - Although only 5% of steady-state peripheral blood (PB) CD34+ cells were found to express chemokine receptor CXCR4, 45% of the cells became CXCR4+ after incubation at 37 degrees C for 4 hours. In contrast, there were no remarkable differences between PB CD34+ cells before and after the 37 degrees C incubation in their expression of selectin ligand, VLA-4, and VLA-5 or in their affinity for VCAM-1 or fibronectin. This increase in CXCR4 expression level was inhibited by the addition of brefeldin A, actinomycin D, or cycloheximide. When PB CD34+ cells with CXCR4 expression levels enhanced by a 4-hour preincubation at 37 degrees C or bone marrow (BM) CD34+ cells were exposed overnight to stromal cell-derived factor 1 (SDF-1), the expression levels of CXCR4 were greatly reduced, and when SDF-1 was removed, CXCR4 levels were thereafter up-regulated. The reexpressed CXCR4 was able to elicit integrin-dependent migration of hematopoietic progenitor cells. There was no difference in the severe combined immunodeficient mouse repopulating cell activity between PB CD34+ cells with and cells without a 37 degrees C preincubation. PMID- 12894852 TI - Wilms tumor gene peptide-based immunotherapy for patients with overt leukemia from myelodysplastic syndrome (MDS) or MDS with myelofibrosis. AB - The Wilms tumor gene, WT1, is overexpressed not only in leukemias and myelodysplastic syndrome (MDS) but also in various types of solid tumors, including lung and breast cancer, and the WT1 protein is a tumor antigen for these malignancies. In clinical trials of WT1 peptide-based cancer immunotherapy, patients with overt leukemia from MDS or MDS with myelofibrosis were injected intradermally with 0.3 mg of an HLA-A*2402-restricted, 9-mer WT1 peptide emulsified with Montanide ISA51 adjuvant. Only a single dose of WT1 vaccination resulted in an increase in WT1-specific cytotoxic T-lymphocytes, which was followed by a rapid reduction in leukemic blast cells. Severe leukopenia and local erythema at the injection sites of WT1 peptide were observed as adverse effects. These results have provided us with the first clinical evidence suggesting that WT1 peptide-based immunotherapy is an attractive treatment for patients with leukemias or MDS. PMID- 12894853 TI - Clinical effects of infusing anti-Epstein-Barr virus (EBV)-specific cytotoxic T lymphocytes into patients with severe chronic active EBV infection. AB - Immune cell therapy with autologous Epstein-Barr virus (EBV)-specific cytotoxic T lymphocytes (CTLs) or lymphokine-activated killer (LAK) cells was performed in 2 adults with severe chronic active EBV infection (SCAEBV). The patient in case 1, who had complications of pancytopenia, high fever, and massive splenomegaly, was treated with 13 doses of LAK cell infusion followed by 4 doses of autologous CTL infusion. The patient in case 2, who had liver dysfunction due to natural killer cell-type infection, was treated with 4 doses of autologous CTL infusion. In case 1, the LAK cell infusions were effective in lowering the viral load and improving several biochemical parameters (lactate dehydrogenase, soluble interleukin 2 receptor) and resulted in complete amelioration of the high fever. Subsequent infusions of autologous CTLs reduced the viral load only temporarily and were accompanied by an increase in frequency of EBV-specific T-cells in the blood. However, the patient's main problem of pancytopenia was not resolved. In case 2, infusion of autologous CTLs did not improve the patient's hepatic dysfunction or viral load but caused a significant increase in autoantibody levels. Thus the effect of auto-CTL treatment was limited or deteriorative in SCAEBV patients. PMID- 12894854 TI - Successful reduced-intensity stem cell transplantation from an HLA haploidentical 3-loci-mismatched donor on the basis of fetomaternal microchimerism in a patient with advanced acute myeloid leukemia. AB - A 31-year-old woman with advanced acute myeloid leukemia underwent non-T-cell depleted (TCD) peripheral blood stem cell transplantation (PBSCT) with a reduced intensity conditioning regimen. The donor was an HLA haploidentical 3-loci mismatched complementary sibling who had not inherited maternal HLA antigens. Long-term fetomaternal microchimerism was detected by nested polymerase chain reaction with specific primer typing. Graft-versus-host disease (GVHD) prophylaxis consisted of tacrolimus with minidose methotrexate. Durable engraftment was achieved without severe acute GVHD, and complete remission was obtained. Thus non-TCD HLA haploidentical reduced-intensity PBSCT based on fetomaternal immunological tolerance appears to be feasible. Our results have important implications in the selection of alternative donors and conditioning regimens for allogeneic hematopoietic stem cell transplantation. PMID- 12894855 TI - Autologous recovery with protractedly undetectable donor chimerisms can precede stable donor engraftment in a nonmyeloablative cord blood transplant. AB - Low levels of donor chimerism on day 28 after hematopoietic stem cell transplantation are highly predictive of a subsequent rejection of donor cells A recent report on nonmyeloablative transplants has demonstrated that patients who engrafted after transplantation had at least 10% donor cells detectable at this time point. A patient is described who received a nonmyeloablative umbilical cord blood transplant and experienced subsequent full engraftment of donor cells despite the absence of donor chimerism at 4 weeks posttransplantation. PMID- 12894856 TI - Eosinophilic colitis in a patient with acute myeloid leukemia after allogeneic bone marrow transplantation. AB - Eosinophilic colitis is a rare inflammatory disease characterized by eosinophilic infiltration of the colon and peripheral blood eosinophilia. We report on a case of eosinophilic colitis in a 29-year-old woman with acute myeloid leukemia following allogeneic bone marrow transplantation from her HLA-identical sister. To our knowledge, eosinophilic colitis has rarely been reported in association with allogeneic bone marrow transplantation. PMID- 12894857 TI - Five novel and four recurrent point mutations in the antithrombin gene causing venous thrombosis. AB - We analyzed the antithrombin (AT) gene in 9 unrelated Japanese patients with thrombotic disease. All 7 exons, the splice junctions, and the 5'-flanking region of the AT gene were amplified by polymerase chain reaction and sequenced directly. Nine different point mutations, all in the heterozygous state, were identified. Five novel (M-32T, M89K, L146H, Q159X, and L409P) and 2 previously reported (R132X and R359X) point mutations were identified in patients with type 1 deficiency. Two different missense mutations, R393C and R393H, located in the protease reactive site were detected in patients with type 2 deficiency. No other sequence abnormalities in the AT gene were detected by direct sequencing. None of the mutations was present in 100 alleles from 50 unrelated Japanese control subjects Although type 1 deficiency was diagnosed in patient 7 on the basis of approximately 50% AT antigen and activity levels, the data indicated that the novel L409P mutation is a type 2 pleiotropic effects (PE) deficiency because its location in the C-terminal portion of the reactive site is similar to the locations of reported PE type mutations, and it is highly conserved among other serpins. PMID- 12894858 TI - Racial background is a determinant factor in the maintenance dosage of warfarin. AB - Warfarin is a drug commonly used in the prevention of thromboembolic events. There have been reports suggesting that racial background may influence warfarin dose requirements. Malaysia is a multiracial country in which there are 3 major races, Malay, Chinese, and Indian. We examined 100 patients from our hospital on stable maintenance doses of warfarin, with international normalized ratio (INR) of 2.0 to 3.5. We found that the mean warfarin dose for Indian patients (n = 19) was 6.9 mg, for Chinese patients (n = 55) was 3.6 mg, and for Malay patients (n = 26) was 3.2 mg. The results showed that the Indian patients required a statistically significantly higher warfarin dose than did patients of the other 2 races (P < .0005). Age was also found to affect the daily warfarin maintenance dose. PMID- 12894859 TI - Effects of nuclear-presenting-macrophage colony-stimulating factor on the process of malignancy. PMID- 12894860 TI - Boston University Doctor of Science Degree Program: clinical doctorate in audiology. AB - This article describes the development of the Boston University Clinical Doctoral Program in Audiology dating from its inception in 1985 to the present. The issues surrounding the establishment of the clinical doctorate, entering the program, and the curriculum are discussed. The unique features of this program and associated resources available within the clinical and scientific community of the greater Boston area are highlighted. PMID- 12894861 TI - The AuD Degree program at Central Michigan University. AB - Central Michigan University's on-campus AuD program admitted its first class of students in 1994. The article discusses its prerequisites, its faculty and staff, and its unique aspects. In addition, CMU's pioneering Web-based AuD program, specifically designed for clinicians with at least 5 years of professional experience, is described. PMID- 12894862 TI - Dehiscence of bone overlying the superior semicircular canal as a cause of an air bone gap on audiometry: a case study. AB - Dehiscence of bone overlying the superior semicircular canal can result in a syndrome of vertigo and oscillopsia induced by loud noises or by maneuvers that change middle ear or intracranial pressure. Patients with this disorder can also experience a heightened sensitivity to bone-conducted sounds in the presence of normal middle ear function. High-resolution CT scans of the temporal bones demonstrate the dehiscence. The authors describe a patient with bilateral superior canal dehiscence who had bilateral low-frequency conductive hearing loss, normal middle ear function, intact acoustic reflexes, and intact vestibular evoked myogenic potentials. These findings would not be expected on the basis of a middle ear cause of the conductive hearing loss. A high-resolution CT scan of the temporal bones in this patient revealed bilateral superior canal dehiscence. Normal acoustic immittance findings in the presence of conductive hearing loss should alert clinicians to the possibility of inner ear cause of an air-bone gap due to superior canal dehiscence. PMID- 12894863 TI - Using talking lights illumination-based communication networks to enhance word comprehension by people who are deaf or hard of hearing. AB - This article details a new method that has been developed to transmit auditory and visual information to people who are deaf or hard of hearing. In this method, ordinary fluorescent lighting is modulated to carry an assistive data signal throughout a room while causing no flicker or other distracting visual problems. In limited trials with participants who are deaf or hard of hearing, this assistive system, combined with commercial voice recognition software, showed statistically significant improvement in sentence recognition compared to recognition of audio-only or audio-plus-speech-reading stimuli. PMID- 12894864 TI - Wanted: a national standard for early hearing detection and intervention outcomes data. AB - The Joint Committee on Infant Hearing (JCIH, 2000) has presented principles and guidelines for universal newborn hearing screening and early hearing detection and intervention (EHDI). The guidelines describe the need for a national data set for early hearing detection and intervention. The guidelines fail to provide the specific constructs for such a data set. To the authors' knowledge, no nationally proposed uniform data structure exists to capture EHDI services' outcome metrics. This article presents a proposed newborn hearing screening and EHDI data model. This model was developed to record EHDI outcomes data from Military Health System birthing centers. The data are to be collected for tracking implementation of Healthy People 2010 goals related to newborn hearing screening and EHDI programs within the Military Health System. In this article, the authors use the T. Helfer, A. Shields, and K. Gates (2000) methods to model a uniform structure for collection of newborn hearing screening and EHDI data. They also discuss expansion of the data model for application to public health reporting of EHDI outcomes in the civilian sector to include integration of Census Bureau demographic data and geographic information system data to further enhance the research value of these EHDI outcomes data. They offer the data model with the intention of supporting national research efforts for studying the efficacy of EHDI programs and to help establish a national evidence-based practice database for such programs. PMID- 12894865 TI - Factor analysis of the Tinnitus Handicap Inventory. AB - The Tinnitus Handicap Inventory (THI; C. W. Newman, G. P. Jacobson, & J. B. Spitzer, 1996) is widely used in a clinical context to assess tinnitus-related self-reported handicap and to report treatment outcomes. Test-retest reliability has been reported to be high, and high convergent validity with other measures of tinnitus distress has been reported. Factor analysis of the subscales of the THI has previously been reported for a Danish translation, but not for the English version. In the present study, the THI was factor analyzed using data collected from a group of 80 clinical tinnitus patients and 116 patients with unilateral vestibular schwannoma. The factor analysis yielded strong support for a unifactorial structure of the scale, with a majority of items loading on the first factor, and high internal consistency of the total score. The authors recommend use of the total score in research and in clinical practice. PMID- 12894866 TI - Visual reinforcement audiometry using digital video disc and conventional reinforcers. AB - Visual reinforcement audiometry (VRA) is a test procedure routinely used to evaluate hearing in infants and young children (6 months to 2 years). Most research and current clinical practice uses flashing lights and/or animated toys to provide reinforcement to a child during VRA. New technology capable of generating a moving video image is now available for providing visual reinforcement to infants during VRA testing. It is reasonable to expect that video images, with presumed greater novelty and complexity, would be more interesting and rewarding to children than conventional, animated mechanical toy reinforcers. On the other hand, in today's society, children are frequently exposed to video images in the home and elsewhere. Therefore, three-dimensional animated toys may present with greater novelty than video images. The purpose of this study was to compare auditory localization behavior, as defined by the number of head turn responses until habituation, during VRA with 2-year-old children using two types of reinforcers: (a) moving images generated by a digital video disc player/monitor and (b) a conventional, animated mechanical toy. Twenty children were selected randomly from a total group of 40 and tested using conventional reinforcement. The remaining 20 children were tested using video reinforcement. The average number of head turn responses prior to habituation was approximately 15 for the video-reinforced group and approximately 11 for the conventional toy-reinforced group, suggesting that during VRA a video image may be more reinforcing than a conventional animated toy. PMID- 12894867 TI - The effects of changes in head angle on auditory and visual input for omnidirectional and directional microphone hearing aids. AB - Improving the signal-to-noise ratio (SNR) for individuals with hearing loss who are listening to speech in noise provides an obvious benefit. Although binaural hearing provides the greatest advantage over monaural hearing in noise, some individuals with symmetrical hearing loss choose to wear only one hearing aid. The present study tested the hypothesis that individuals with symmetrical hearing loss fit with one hearing aid would demonstrate improved speech recognition in background noise with increases in head turn. Fourteen individuals were fit monaurally with a Starkey Gemini in-the-ear (ITE) hearing aid with directional and omnidirectional microphone modes. Speech recognition performance in noise was tested using the audiovisual version of the Connected Speech Test (CST v.3). The test was administered in auditory-only conditions as well as with the addition of visual cues for each of three head angles: 0 degrees, 20 degrees, and 40 degrees. Results indicated improvement in speech recognition performance with changes in head angle for the auditory-only presentation mode at the 20 degrees and 40 degrees head angles when compared to 0 degrees. Improvement in speech recognition performance for the auditory + visual mode was noted for the 20 degrees head angle when compared to 0 degrees. Additionally, a decrement in speech recognition performance for the auditory + visual mode was noted for the 40 degrees head angle when compared to 0 degrees. These results support a speech recognition advantage for listeners fit with one ITE hearing aid listening in a close listener-to-speaker distance when they turn their head slightly in order to increase signal intensity. PMID- 12894868 TI - Immunomodulatory effects of HMG-CoA reductase inhibitors. AB - 3-Hydroxy-3-methyl-glutaryl coenzyme A (HMG-CoA) reductase inhibitors, or statins, are competitive inhibitors of the rate-limiting enzyme in cholesterol synthesis. Several clinical trials have shown a marked reduction in cholesterol levels associated with decreased cardiovascular mortality in patients treated with statins. However, more recent observations have suggested that the clinical benefits of statins may be, at least in part, independent of the effect of statins on cholesterol synthesis. These so-called pleiotropic or cholesterol independent effects of statins could be the result of reduction in the formation of intermediaries in the mevalonate pathway as statins, by inhibiting L-mevalonic acid synthesis, also prevent the production of isoprenoids in the cholesterol biosynthetic pathway. Isoprenoids serve as important lipid attachments for the posttranslational modification of a variety of proteins such as small GTP-binding proteins of the Ras superfamily implicated in intracellular signaling. The list of different pleitropic effects of statins is still growing and includes, among others, direct effects of statins on modulating endothelial function, decreasing oxidative stress and, more recently, anti-inflammatory and immunomodulatory actions of statins. For instance, statins decrease T cell activation, the recruitment of inflammatory cells into atherosclerotic lesions, and inhibit IFN gamma expression of MHC II on antigen-presenting cells. This review article summarizes the anti-inflammatory and immunomodulatory effects of statins and thus provides a new rationale to use statins as a new class of immunosuppressive agents. PMID- 12894869 TI - Experimental models of inflammatory bowel disease. AB - The etiology and pathogenesis of inflammatory bowel disease (IBD) remains unsolved, but improved experimental models of enterocolitis have led to progress. Intestinal inflammation and experimental IBD can be induced by chemical or dietary factors or by microbial products. Many animal models of IBD can be used to evaluate new anti-inflammatory drugs. These models, however, usually demonstrate acute, self-limiting colitis. The spontaneous colitis models developed in the cotton-top tamarin monkey and the C3H/HeJBir mouse mimic more features of human IBD. Inflammation is chronic and is under genetic control. The differential genetic susceptibility of inbred rat strains to chronic inflammation have been exploited. Lewis rats injected with bacterial products, peptidoglycan polysaccharide or indomethicin develop chronic relapsing enterocolitis, whereas closely related Buffalo or Fisher rat strains develop only transient inflammation. These models are also useful to test the specific inhibition of inflammatory mediators and target molecules. Over-expression (transgenic) or deletion (knockout) of specific genes have led to the development of rodent models of spontaneous colitis. Inflammation arises from a number of mutations of immunomodulatory molecules, supporting the concept of genetic heterogeneity for IBD. The results obtained from experimental models have generated new hypotheses, expanded human studies, and suggested novel forms of therapy for IBD patients. PMID- 12894870 TI - Wegener's granulomatosis--autoimmunity to neutrophil proteinase 3. AB - Wegener's granulomatosis (WG) is a small-vessel vasculitis associated with various clinical manifestations, among which the most common are respiratory tract disease and glomerulonephritis leading to renal failure. The pathogenesis of vascular injury in WG is ascribed to antineutrophil cytoplasmic antibodies (ANCA) directed mainly against proteinase 3 (PR3), an enzyme from neutrophil granules. The reasons for the breakdown of self tolerance to PR3 are unknown, and together with the molecular mechanisms underlying this immunoinflammation, are the subject of research. Standard treatment of WG consists of cyclophosphamide and corticosteroids. In patients resistant to this therapy or with refractory disease, some alternative strategies involving tumor necrosis factor blockade, polyclonal antithymocyte globulin or monoclonal anti-T cell antibodies are applied. PMID- 12894871 TI - CSF-1 as a regulator of macrophage activation and immune responses. AB - Macrophage activation is a key determinant of susceptibility and pathology in a variety of inflammatory diseases. The extent of macrophage activation is tightly regulated by a number of pro-inflammatory cytokines (e.g. IFN-gamma, IL-2, GM CSF, IL-3) and anti-inflammatory cytokines (e.g. IL-4, IL-10, TGF-beta). Macrophage colony-stimulating factor (CSF-1/M-CSF) is a key differentiation, growth and survival factor for monocytes/macrophages and osteoclasts. The role of this factor in regulating macrophage activation is often overlooked. This review will summarize our current understanding of the effects of CSF-1 on the activation state of mature macrophages and its role in regulating immune responses. PMID- 12894872 TI - Immunosuppressive agents and cytomegalovirus infection. AB - Cytomegalovirus (CMV) infection is the major infectious complication observed after organ transplantation. As rejection episodes always occur in allograft transplanted recipients, various kinds of immunosuppressive agents are used to control such rejection episodes. Among the commonly used immunosuppressive agents, anti-pan-T cell polyclonal and monoclonal antibody are known to increase the risk of viral infections. New immunological techniques have recently been developed to measure CMV-specific CD4 and CD8 cells by flow cytometry. Using the techniques, high frequencies of specific CD4 and CD8 T cells have been shown to be required to survey the CMV (re)activation in the persistent/latent phase of CMV infection. An excessive T cell depletion by OKT3 would deplete such surveying T cells, thus resulting in the occurrence of CMV-associated diseases. PMID- 12894873 TI - 17Beta-estradiol treatment profoundly down-regulates gene expression in spinal cord tissue in mice protected from experimental autoimmune encephalomyelitis. AB - It is now well documented that experimental autoimmune encephalomyelitis (EAE) can be effectively prevented by estrogen therapy. Previously, we identified a limited set of genes that were altered in spleens of mice protected from EAE by 17beta-estradiol (E2) treatment. As a continuation of these studies, we present here transcriptional changes in genes expressed in spinal cord tissue. The Affymetrix microarray system was used to screen more than 12,000 genes from E2 treated double transgenic (BV8S2 and AV4) female mice protected from EAE vs. control mice with severe EAE. We found that estrogen therapy had a profound inhibitory effect on the expressions of many immune-related genes in spinal cords. Estrogen significantly affected the transcription of 315 genes, 302 of which were down-regulated and only 13 that were up-regulated by > or = 2.4 fold. A number of genes encoding the histocompatibility complex, cytokines/receptors, chemokines, adhesion molecules, and signal transduction proteins were strongly down-regulated (> 20 fold) in estrogen-treated mice to levels similar to those of the spinal cord tissue from unmanipulated mice. The identification of genes with altered expression patterns in the spinal cords of estrogen-treated mice provides unique insight into the process that ultimately results in protection against EAE. PMID- 12894874 TI - Diagnostic value of pancreatic elastase-1 in human acute pancreatitis. AB - The diagnosis of acute pancreatitis (AP) is usually confirmed by a significant increase of the serum amylase and/or lipase level. However, serum pancreatic elastase-1 (pEla-1) was found to be a more sensitive diagnostic marker in AP, when assayed by the radioimmunoassay procedure. We analyzed the serum concentration of pEla-1, measured by the ELISA technique in 46 patients with AP and in a control group of 12 healthy volunteers. On admission (day 1) we found significantly higher pEla-1 levels in patients with AP than in the controls. During the following days, the concentration of pEla-1 rapidly decreased to nearly undetectable values on the 3rd day. There was no significant difference between patients with mild and severe AP nor those of different etiology. We suggest that pEla-1 has little diagnostic value and does not provide additional information to that of the less expensive and more widely available serum amylase and lipase. PMID- 12894875 TI - Lack of association between an exon 1 CTLA-4 gene polymorphism A(49)G and multiple sclerosis in a Polish population of the Lower Silesia region. AB - Multiple sclerosis (MS) a chronic inflammatory demyelinating disease of the central nervous system is believed to have a T cell-mediated autoimmune etiology. The cytotoxic T lymphocyte antigen 4 (CTLA-4) gene is a strong candidate for the involvement in autoimmune diseases because CTLA-4 plays an important role in the downregulation of the early and late stages of T cell activation and the maintenance of peripheral T cell tolerance. To examine the genetic association of the CTLA-4 gene locus with MS, we analyzed an exon 1 CTLA-4 gene polymorphism A(49)G in 102 unrelated Polish MS patients in the Lower Silesia region and 101 age- and sex-matched healthy subjects. The distribution of CTLA-4 exon 1 A(49)G genotype, phenotype and allele frequencies did not differ between patients with MS and healthy subjects. PMID- 12894876 TI - Antitumor effects of monoclonal antibody Fab' fragment-containing immunoconjugates. AB - OBJECTIVE: Using monoclonal antibody (mAb) Fab' fragment to develop mAb immunoconjugates for cancer. METHODS: Fab' fragment of mAb 3A5 was prepared by digestion of the antibody with pepsin and then reduced by dithiothreitol (DT), while Fab' fragment of mAb 3D6 was obtained by digestion of the antibody with ficin and subsequently reduced by beta-mercaptoethanol. The conjugation between Fab' fragment and pingyangmycin (PYM), an antitumor antibiotic, was mediated by dextran T-40. Immunoreactivity of Fab'-PYM conjugates with cancer cells was determined by ELISA, and the cytotoxicity of those conjugates to cancer cells was determined by clonogenic assay. Antitumor effects of the Fab'-PYM conjugates were evaluated by subcutaneously transplanted tumors in mice. RESULTS: The molecular weight of Fab' fragment was approximately 53 kD, while the average molecular weight of Fab'-PYM conjugate was 170 kD. The Fab'-PYM conjugates showed immunoreactivity with antigen-relevant cancer cells and selective cytotoxicity against target cells. Administered intravenously, Fab'-PYM conjugates were more effective against the growth of tumors in mice than free PYM and PYM conjugated with intact mAb. CONCLUSION: Fab'-PYM conjugate may be capable of targeting cancer cells and effectively inhibiting tumor growth, suggesting its therapeutic potential in cancer treatment. PMID- 12894877 TI - Cloning and characterization of human ubiquitin binding enzyme 2 cDNA. AB - OBJECTIVE: To clone and identify the gene encoding human ubiquitin binding enzyme 2 and study its expression pattern. METHODS: According to the sequence of human EST, which is highly homologous to the mouse ubiquitin binding/conjugating enzyme (E2), primers were synthesized to screen the human fetal brain cDNA library. The gene was analyzed by bioinformatics technique and its expression pattern was studied by using multiple-tissue Northern blot. RESULTS: Two cDNA clones encoding human ubiquitin conjugating enzyme have been isolated and identified. Both containing the ubiquitin conjugating domain, the 2 cDNA clones are 88% identical in amino acid sequences and splicing isoforms to each other only with an exon excised to form the short sequence. They belong to a highly conserved and widely expressed E2 enzyme family. Northern blot shows that they are expressed exclusively in adult human heart, placenta, and pancreas but no transcripts can be detected in brain, lung, liver, skeletal muscle or kidney. CONCLUSIONS: The gene encoding human ubiquitin binding enzyme is expressed under temporal control. As a key enzyme in the degradation of proteins, ubiquitin conjugating enzymes play a central role in the expression regulation on the level of post translation. PMID- 12894878 TI - In vitro analysis of tau phosphorylation sites and its biological activity. AB - OBJECTIVE: To explore the association between the abnormal phosphorylation sites found in Alzheimer disease (AD) tau and the inhibition of its biological activity. METHODS: Ultracentrifugation, chromatography, manual Edman degradation and autosequence techniques were used to prepare and phosphorylate human recombinant tau, isolate and purify 32P tau peptides and determine phosphorylation sites. RESULTS: Phosphorylation of tau by casein kinase 1 (CK-1), cyclic AMP-dependent protein kinase (PKA) and glycogen synthetase kinase 3 (GSK 3) separately inhibited its biological activity and the inhibition of this activity by (CSK-3 was significantly increased if tau was prephosphorylated by CK 1 or PKA. The most potent inhibition was seen by a combined phosphorylation of tau with PKA and GSK-3. The treatment of tau by PKA and GSK-3 combination induced phosphorylation of tau at Ser-195, Ser-198, Ser-199, Ser-202, Thr-205, Thr-231, Ser-235, Ser-262, Ser-356, Ser-404, whereas Thr-181, Ser-184, Ser-262, Ser-356 and Ser-400 were phosphorylated by GSK-3 alone under the same condition. CONCLUSION: Phosphorylation of tau by PKA plus GSK-3 at Thr-205 might play a key role in tau pathology in AD. PMID- 12894879 TI - Detection of B lymphoma cells undergoing apoptosis by Annexin-V assay. AB - OBJECTIVE: To quantitatively analyze apoptotic and secondary necrotic cells under apoptosis conditions. METHODS: The cells of Burkitt lymphoma (BL) cell line Raji were incubated with 1.0 micromol/L dexamethasone (DEX) for 2, 4 and 8 h respectively, then stained with Annexin V-FITC (fluorescein isothiocyanate conjugated) which was used to detect the exposed phosphatidylserine (PS) on the epimembrane resulting from a loss of phospholipid asymmetry in the early stage of apoptosis, and also stained with propidium iodide (PI) which allows analysis of secondary necrotic cells related with cell membrane and DNA damage that probably representlate stage of apoptosis, then apoptotic cells were quantified by flow cytometry (FCM). Furthermore, Annexin+ /PI- and Annexin+ /PI+ cells were sorted by fluoresence-activated cell sorter (FACS), and identified by electron microscopy (EM) and DNA gel electrophoresis. RESULTS: The percentage of apoptotic cells was found to increase with the incubation time (r = 0.97). This method was sensitive with low detection limit (0.02%), and was reproducible with low coefficient variance (CV) (4.2%). Meanwhile, the Annexin+ /PI- and Annexin+ /PI+ cells were identified as apoptotic and necrotic cells under EM, and DNA extracted from the Annexin+ /PI- cells was characterized by "ladder pattern". CONCLUSIONS: Annexin-V assay is a specific, sensitive, accurate, reproductive and quantitative method for analyzing apoptotic cells. PMID- 12894880 TI - The effect of all-trans retinoic acid on gap junctional intercellular communication and connexin 43 gene expression in glioma cells. AB - OBJECTIVE: To illuminate the regulating effect of all-trans retinoic acid (ATRA) on gap junctional intercellular communication (GJIC) and connexin 43 (Cx43) gene expression in glioma cells, which is tissue- and organ-specific. METHOD: Rat C6 glioma cells were exposed to ATRA at a concentration of 1, 10, 100 micromol/L and the GJIC function of the cells was examined with scrape-loading dye transfer assay 24 hours, 48 hours and 72 hours after ATRA treatment. The effect of ATRA on Cx43 gene expression was measured with semiquantitative reverse transcription polymerase chain reaction (RT-PCR) 24 hours after ATRA exposure. RESULTS: The GJIC function of C6 glioma cells was significantly increased by ATRA at each concentration applied. The dye passed 4 to 5 rows of cells from the scraping edge in ATRA treated cells, but only 1 or 2 rows in the control. The augment effect was observed 24 hours after each concentration ATRA treatment, and lasted till 72 hours after treatment with 1 micromol/L and 10 micromol/L ATRA. Forty-eight hours after exposed to 100 micromol/L ATRA, the enhancement of GJIC was less obvious. There was no significant increase induced by ATRA on the transcription of Cx43 gene, as demonstrated by semiquantitative RT-PCR. CONCLUSION: ATRA turned out to be a potent enhancer on GJIC function in C6 glioma cells, andthe enhancement effect was most probable at post-transcriptional level. PMID- 12894881 TI - New designed HMBA agents as inducers of erythroleukemia cell differentiation. AB - OBJECTIVE: Searching for more potent and less toxic HMBA-related agents. METHODS: Human erythroleukemia cell K562, murine erythroleukemia cell (MEL) and its sub line MEL DS19 were used as target cells to select a cell line which is the most sensitive to HMBA, then analyzed the activity of inducing differentiation of two new designed HMBA derivatives: HMBPA [hexamethylenebi (3-pyridin) amide] and Co HDTA (ethylenediaminetetra acetic acid cobalt) using cell biology, cytochemical and molecular biology techniques. RESULTS: We found that the MEL DS19 cells were most sensitive to HMBA (benzidine positive, B+ approximately 76%). Co-HDTA can inhibit the growth of MEL DS19, but induces differentiation just in a small population (B+ 2% approximately 4.5%). Between 0.02 approximately 5 micromol/L, HMBPA induces 3% approximately 8% cells committed to differentiation with little inhibition of cell proliferation. 1 micromol/L HMBPA and 2 mmol/L HMBA together, can obviously increase the percentage of differentiated cell (B+ approximately 72%), inhibit DNA synthesis and accelerate beta-globin transcription. CONCLUSION: The new HMBA derivatives may provide potential cancer differentiation inducers. PMID- 12894882 TI - Clinical and pathological manifestations of patients with antineutrophil cytoplasmic autoantibodies directed against proteinase 3 or myeloperoxidase. AB - OBJECTIVE: To compare the clinical and pathological manifestations of patients with antineutrophil cytoplasmic autoantibodies (ANCA) directed against proteinase 3 (anti-PR3) or myeloperoxidase (anti-MPO). METHODS: One hundred and forty patients with ANCA were detected for anti-PR3 and anti-MPO by ELISA. The clinical features at presentation, histopathological characteristics and outcome of all patients who were tested positive for anti-PR3 or anti-MPO were analysed. RESULTS: In anti-PR3 group (n = 21), 16 cases (76.2%) had systemic vasculitis, in which Wegener's granulomatosis prevailed (13 cases, 61.9%). In anti-MPO group (n = 31), 19 cases (61.3%) were diagnosed as systemic vasculitis and 12 cases (38.7%) as microscopic angiitis. For vasculitic patients with anti-PR3 and anti MPO, the disease duration at diagnosis was 9.6 +/- 2.0 m and 4.4 +/- 0.9 m respectively, P < 0.05; vasculitis activity index (BVAS) and mean number of affected organ were 22.5 +/- 2.1, 5.0 +/- 0.4 and 25.1 +/- 1.7, 4.8 +/- 0.4 respectively, P > 0.05; upper respiratory tract, eye and joint involvements were 11(68.8%), 7(43.8%), 11(68.8%) and 7(36.8%), 2(10.5%), 5(26.3%) respectively, P < 0.05. Although there was no statistical difference in renal involvement between these two groups, patients with serum creatine > 500 micromol/L were more commonly seen in anti-MPO group than in anti-PR3 group, which were 8(42.1%) and 2(12.5%) respectively, P < 0.05. Ten relapses were seen in anti-PR3 group and only 2 in anti-MPO group, but the acute mortality rate in anti-MPO group (5/19, 27.4%) was much higher than that in anti-PR3 group (1/16, 6.3%). CONCLUSIONS: Anti-PR3 and anti-MPO occurred mainly in systemic vasculitis. A large divergence was seen in the disease spectrum between patients with anti-PR3 and those with anti-MPO. In particular, upper respiratory tract, eye and joint involvements, granuloma formation and relapse were more prominent in anti-PR3 patients. By contrast, the anti-MPO patients had a more acute disease onset, more rapid progressive renal involvement and a higher acute mortality rate. PMID- 12894883 TI - Procoagulant effects of thrombolytic therapy in acute myocardial infarction. AB - OBJECTIVE: To examine the procoagulant effects of thrombolytic agent on hemostasis and study the role of hemostatic markers as predictors of clinical outcomes. METHODS: In the present study, eighteen patients with acute myocardial infarction (AMI) received 1.5 or 2.0 million U nonspecific urokinase (UK), or 70 approximately 80 mg fibrin-specific recombinant tissue plasminogen activator (rt PA) and did not use heparin until 8 hours after intravenous injection of the above agents. Eight patients with AMI and without thrombolytic therapy were enrolled as controls. Coagulant and thrombolytic activity markers included thrombin-antithrombin III complex (TAT), D-dimer, fibrinogen (Fg), FMPV/Amax. All markers were determined before, immediately, 1, 2, 4 and 8 hours after the administration of thrombolytic agents respectively. RESULTS: Molecular marker of thrombin generation--TAT showed an activated coagulant state immediately after thrombolytic therapy. Level of TAT showed no significant changes between every two observed phases in controls. However, level of TAT increased significantly from 4.95 +/- 1.75 microg/L ( 4.63 +/- 1.37 microg/L) to 14.71 +/- 3.31 microg/L (14.25 +/- 2.53 microg/L) before and immediately after administration of thrombolytic agents UK (or rt-PA). There was significant difference between level of serum TAT of patients with and without thrombolytic therapy (P < 0.05). Patients achieving clinical reperfusion had lower TAT level than those failing in thrombolytic therapy, and higher FMPV/Amax level than controls. D-dimer, a surrogate of thrombolytic activity increased markedly and Fg significantly declined after thrombolytic therapy (P < 0.05). CONCLUSIONS: Thrombin generation occurred in plasma in response to excess fibrinolysis induced by thrombolytic therapy. Both urokinase and rt-PA had procoagulant action. This transient activation of the coagulant system might contribute to early reocclusion. These data provided the theoretical support for simultaneous administration of anticoagulant therapy with thrombolytic agents. These results also suggested that TAT might be useful in predicting clinical outcomes of patients treated with thrombolytic therapy for AMI. PMID- 12894884 TI - Relation of hypertension to diabetic nephropathy in patients with non-insulin dependent diabetes mellitus--a pair-matched case-control study. AB - OBJECTIVE: To masess the role of hypertension and family history of hypertension in the development of nephropathy in patients with non-insulin-dependent diabetes mellitus (NIDDM). METHODS: A retrospective analysis was done on 2 groups of NIDDM patients, one group without proteinuria (urine protein < 300mg/24h, n = 106) and the other group with proteinuria (urine protein > or = 500mg/24h, n = 106). The 2 groups were matched by age (< or = +/- 3yrs), sex, ethnic and resident place. Some information of these subjects including demographic; history of disease, family history of diseases, lifestyle and behavior style variables was obtained by questionnaire; some variables were measured, including systolic blood pressure (SBP), diastolic blood pressure (DBP), fasting blood glucose (FBG), quantity of protein in 24h urine. Then conditional logistic regression analysis was performed. RESULTS: Some factors, including history of hypertension, longer duration of hypertension, higher levels of the past highest SBP and DBP, were independently associated with the occurrence risk of diabetic nephropathy (DN). Their corresponding odd ratios (OR) with 95% confidence intervals (CI) were 2.00(1.17 approximately 3.43), 1.25(1.08 approximately 1.46), 1.38(1.15 approximately 1.66), and 1.33(1.09 approximately 1.62) respectively, but family history of hypertension was not significantly associated with the development of DN. When/the above-mentioned relations were respectively adjusted by some possible confounding factors, they still existed. CONCLUSIONS: History of hypertension, longer duration of hypertension, higher levels of the past highest SBP and DBP are independent risk factors for DN in Chinese NIDDM patients. PMID- 12894885 TI - Liver transplantation in patients with fulminant hepatitis B: experience in Canton, China. AB - OBJECTIVE: To investigate the effect of orthotopic liver transplantation on fulminant hepatitis B and the preventive efficiency of lamivudine on recurrence of hepatitis B in China. PATIENTS AND METHODS: Ten patients with fulminant hepatitis B received orthotopic liver transplantation under veno-venous bypass. All patients had preoperatively serious jaundice, ascites and coagulopathy, and of whom 7 with encephalopathy, 2 with acute renal failure, and 1 with gastro hemorrhage. RUSULT: Seven of the 10 patients have survived for 3 approximately 18 months, but 3 died of multi-organ failure or recurrence of fulminant hepatitis B. Seven survivors took lamivudine and 6 of them have survived for 3 approximately 18 months without the signs of recurrence of hepatitis B. CONCLUSION: Orthotopic liver transplantation is an effective therapy for fulminant hepatitis B, and lamivudine may prevent recurrence of hepatitis B after transplantation. PMID- 12894886 TI - Comparison of cellulose acetate polymer and electrolytic detachable coils for treatment of canine aneurysmal models. AB - OBJECTIVE: Electrolytic detachable coils (EDC) have been the main embolic materials for intracranial aneurysms. Liquid aneurysmal embolic materials represented by cellulose acetate polymer (CAP) are still in controversy. In this research, the embolization results and pathological reactions after embolization of canine aneurysmal models with EDC or CAP were observed and compared. METHODS: The canine aneurysmal models constructed by anastomosis of venous pouches were randomly grouped. The aneurysms were respectively occluded with CAP and electrolytic detachable coils that was named by Wu electrolytic detachable coil (WEDC) and made by us. Angiogram follow-ups were performed at 24-hour, 2-week, and 2-month after embolization. The occluded aneurysms were dissected in each stage for light microscopic, electron microscopic, and histochemical research. RESULTS: The effect of embolization was significantly better with WEDC than that with CAP (chi2 = 5. 56, P < 0.05). Post-embolized complications such as aneurysm rupture and stenosis of parent arteries could only be found in CAP group. Pathological research showed that CAP mass could packed the aneurysms more densely than coils. Acute chemical damage of aneurysmal wall and inflammatory cell infiltration was prominently found in early stage after CAP-embolization. Organization of thrombus inside aneurysms and formation of endothelial tissue over the orifices of aneurysmal necks could be found in both groups 2 months after embolization. But parts of coils might be exposed outside endothelial layer. CONCLUSIONS: EDC are still the most safe, efficient, and reliable instruments to embolize aneurysm. CAP should be improved further to solve the problem of strong chemical corrosion and difficulty in control before it is widely used. PMID- 12894887 TI - Clinical application of the TRAFIX instrumentation in correcting scoliosis. AB - OBJECTIVE: To evaluate the results of the TRAFIX instrumentation in correcting scoliosis. METHODS: Since October 1997, 47 patients with scoliosis received spinal fixation with the TRAFIX instrumentation at the Peking Union Medical College Hospital. The average age was 14. 3 years (range 10 to 38 years). There were 27 idiopathic cases, 16 congenital cases, 2 cases with Marfan syndrome and 2 with neurofibromatosis. Twelve of the 47 patients underwent anterior release, while 4 patients received the revision approach. The average follow-up time was 26 months (13 -38 months). RESULTS: The measurements of primary coronal deformity before and after surgery were 74 degrees (50 degrees approximately 115 degrees) and 38.7 degrees (11 approximately 95 degrees), respectively. The average curve correction was 54%. The average number of fused segments was 12.5 (7 approximately 17) vertebrae. The distance between the center of apex and the C7 plumb line was 56.8 mm before surgery (25 approximately 107 mm) and 31 mm after surgery (10 approximately 87 mm). CONCLUSION: The TRAFIX instrumentation provides three-dimensional correction with refinement, convenience and reliable fixation. PMID- 12894888 TI - China-made platinum-iridium alloy stent implanted in the coronary of dog. PMID- 12894889 TI - The development of burn surgery in China--a retrospective overview. PMID- 12894890 TI - Study of BRCA1 gene in hereditary breast and ovarian cancer. AB - OBJECTIVE: To investigate the BRCA1 gene in hereditary breast and ovarian cancer, early-onset breast cancer and sporadic ovarian cancer. METHODS: The exons of 2, 11 and 20 of BRCA1 gene were analyzed. Polymerase chain reaction-single strand conformation analysis (PCR-SSCP) and PCR-SSCP combined by restriction enzymes were used to screen for mutations. Mutations were further indentified by sequencing. The loss of heterozygosity (LOH) were also investigated at the BRCA1 genetic loci D17S855 in 10 hereditary ovarian cancer. RESULTS: A insertion mutation was detected in H7. "C" was inserted at nucleotide 797. It would result in truncation of the BRCA1 protein at codon 277. A missense mutation was detected in an early-onset breast cancer (diagnosed at age 24). At nucleotide position 3732, the substitution of a "G" to a "C" in codon 1205 changes a Gly to a Arg. A missense mutation were also detected in three sporadic ovarian cancers. At nucleotide position 2051, the substitution of a "T" to a "G" in codon 644 changes a Cys to a Trp. H3 and H7 patients show LOH. CONCLUSIONS: BRCA1 gene has an important effect in Chinese hereditary breast and ovarian cancer, its effect on early-onset breast cancer and sporadic ovarian cancer are still to be studied. BRCA1 gene is a tumor-suppressor gene. PMID- 12894891 TI - Deletions and point mutations of p16,p15 gene in primary tumors and tumor cell lines. AB - Aberrations of chromosome 9 p21-22 are involved in the genesis of many forms of cancer. The gene p16 and p15 have been assigned to this region. Both p16 and p15 are an inhibitor of cyclin D-cdk4,cyclin D-cdk6 complex and have been implicated in a wide variety of cancer types, including the germline of patients with familial melanoma. In order to investigate and compare the status of p16,p15 gene in primary tumors and cell lines, we examined 357 primary tumors and 29 cell lines derived from diverse tumor types. In addition to analysis of these primary tumors and cell lines, blood specimens from 91 patients either with sporadic multiple cancers or from cancer-prone families were also analyzed. The data showed the following: 1) Homozygous deletions of p16,p15 were comparatively rare and far less common than previously reported, although hemizygous deletions were observed in a significant fraction of many tumor types; 2) the incidence of p16,p15 deletions (either homozygous deletions or heterozygous deletions) varied significantly among different tumor types; 3) most deletions involved in both p16 and p15 genes; 4) sequence variations in the coding sequence of p16,p15 were comparatively rare among these tumor types, though mutations and polymorphisms were identified; 5) some tumors which showed LOH at 9p, containing p16 and p15 gene, did not show deletions or point mutations in the p16,p15 gene. 6) In a subset of retinoblastoma and osteosarcoma where no Rb gene mutations were present a significant fraction was found to contain p16,p15 gene deletions. PMID- 12894892 TI - Inactivation of p16 gene in leukemia. AB - To determine the frequency of p16 gene inactivation in leukemia cells, and to evaluate their value in the prediction of their clinical outcome. Bone marrow or peripheral blood samples from 48 patients with leukemia were examined by multiplex polymerase chain reaction (MPCR) to detect p16 gene homozygous deletion, and restriction enzyme PCR to detect p16 gene methylation. p16 gene inactivation were detected in 10 of the 48 patients (20.4%). They were five patients with p16 homozygous deletion, and five patients with p16 methylation, respectively. p16 gene inactivation correlates with adverse prognosis features. The patients with p16 inactivation had poor response to chemotherapy, and had significantly shorter survival times than the patients in whom p16 gene was preserved (P<0.001). The inactivation of p16 gene play a key role in the pathogenesis and the progression of some leukemia. The detection of p16 gene is reliable prognostic factor that predict shortened survival times. PMID- 12894893 TI - Transcranial Doppler characteristics in persistent vegetative status, locked-in syndrome and brain death. AB - AIM: Comparison of the transcranial Doppler (TCD) characteristics of cerebral circulation in persistent vegetative status (PVS), locked-in syndrome and brain death patients. METHODS: Using TCD ultrasound to detect the flow velocity and waveform patterns of middle cerebral artery (MCA) and basilar artery (BA) in patients with PVS, locked-in syndrome and brain death. RESULTS: The mean velocities of middle cerebral artery (Vmca) and basilar artery (Vba) were 30.0 cm/s and 24.3 cm/s in PVS patients respectively, which decreased 45.0% and 14.4% in comparing with normal value. For patients with locked-in syndrome, Vmca and Vba were 49.7 cm/s and 9.8 cm/s, which decreased 5.0% and 61.7% than the normal value respectively. These results showed that the decrease of anterior circulation was predominant in PVS, and the decrease of posterior circulation was predominant in locked-in syndrome. A unique diastolic reverse flow, short peak systolic wave or undetectable flow signal in middle cerebral artery were predominant in brain death patients, which was completely different from that of either PVS or locked-in syndrome. CONCLUSION: TCD was a valuable tool in distinguishing PVS, locked-in syndrome and brain death patients according to the differences in velocities and patterns of anterior and posterior cerebral arteries. PMID- 12894894 TI - Neutrophilic transient acidification and superoxide production in peritoneal dialysate. AB - AIM: To elucidate whether an inhibited superoxide production (O2-) of neutrophils induced by commercial lactate-based peritoneal dialysates (PDS) could be corrected after a transient intracellular acidosis. METHODS: The intracellular pH ([pHi]) of human neutrophils incubated in PDS was monitored with a spectrofluorometer with a pH-sensitive dye (BCECF-AM). Neutrophilic O2- stimulated by zymosan was determined in PDS with the superoxide dismutase inhibitable ferricytochrome c reduction, using a spectrophotometer. RESULTS: The severe intracellular acidosis induced within 5 min by PDS at an extracellular pH of 5.2 could be promptly and completely recovered by a neutralization of the pH of media. However, O2- by neutrophils exposed to the PDS for as little as 5 min was drastically and persistently inhibited, even the acidic [pHi] of cells had been fully returned for 1 h. CONCLUSIONS: The intracellular acidification of cells in the initial phase could be transient and reversible, but impaired cell functions, at least in part including O2- generating system, might be consistent and irreversible in the early stage of the cellular acidosis in the peritoneal cavity of CAPD patients. The findings above may be of particular importance in both clinic and cell biology. PMID- 12894895 TI - The alteration of the pulmonary artery flow spectrum with pulmonary hypertension. AB - OBJECTIVE AND METHOD: To research the alteration of pulmonary flow spectrum of patients with congenital heart disease and the relation between pulmonary flow spectrum and pulmonary artery pressure, 33 patients with congenital heart disease were examined by echocardiography and catheterization. RESULTS: The result showed that the spectrum peak early occurring in patients with pulmonary hypertension and the degree of the spectrum peak early occurring was related to pulmonary artery pressure positively. The result suggested that when the pulmonary artery systolic pressure was over 100 mmHg, the flow spectrum showed slender pattern usually. CONCLUSION: The slender pattern in pulmonary flow spectrum could be a semi-quantity parameter to predict that the pulmonary artery systolic pressure more than 100 mmHg. PMID- 12894896 TI - Phase-1 studies of malariotherapy for HIV infecton. AB - OBJECTIVE: To determine whether malariotherapy (an old therapy for treatment of neurosyphilis) improves some clinical and laboratory parameters of HIV-positive patients without iatrogenic complications. METHODS: Total 8 asymptomatic HIV-1 positive subjects whose CD4 cell counts were over 250 x 10(6) cells/L were selected for the phase-1 studies of malariotherapy and were intravenously injected Plasmodia vivax to induce artificial malaria. Malaria was terminated with chloroquine after 10 to approximately 20 malarial fever episodes. Cell-bound CD4 levels were measured by APAAP (a solid-phase enzyme essay) and levels of neopterin (NPT), beta-2-microglobulin (B2M), soluble tumor necrosis factor receptor-2 (sTNF-RII), interleukin-2 (IL-2) and HIV P24 antigen were measured by ELISA. Patients were followed up to 24 to approximately 30 months. RESULTS: CD4 levels increased in 5, NPT decreased in 7 of 8 patients; IL-2 increased in 5 of 6 patients after malariotherapy. The total trends of B2M and sTNF-RII basically remained stable. HIV P24 antigen remained undetectable in 6, remained detectably low level in 1 and experienced increase in 1 of 8 patients after malariotherapy. No any severe complications occurred in all 8 patients. CONCLUSIONS: The results indicate that malariotherapy basically is safe for HIV infection and it seems that the therapy improves some immunological parameters of HIV patients. PMID- 12894897 TI - Diagnosis and treatment of the malignant gestational trophoblastic tumor with pulmonary metastasis complicated with pulmonary tuberculosis. AB - OBJECTIVE: To evaluate the diagnosis and treatment for malignant gestational trophoblastic tumor (MGTT) with pulmonary metastasis complicated with pulmonary tuberculosis. METHODS: To analyze ten cases of MGTT with pulmonary metastasis complicated with pulmonary tuberculosis in our hospital from 1980 to 1997 retrospectively. RESULTS: From the x-ray film, there are great resemblances between MGTT with pulmonary metastasis and pulmonary tuberculosis. Of 10 patients, 7 of them were examined out pulmonary tuberculosis during the chemotherapy of MGTT. Pulmonary tuberculosis appeared six months before chemotherapy in three cases. All of the patients were treated with multiagent chemotherapy. Seven patients achiceved a complete remission, 2 patients developed drug resistance and died of cerebral haemorrhage and cerebral herniation, 1 woman who had achieved a complete remission from MGTT for 14 months died of miliary tuberculosis. CONCLUSION: It is very important to make differential diagnosis of the MGTT with pulmonary metastasis complicated with pulmonary tuberculosis. Trying to avoid excessive anti-tumor treatment owing to mistake pulmonary tuberculosis for pulmonary metastasis, and avoiding missing an opportunity of anti-tuberculosis treatment because of missed diagnosis should be emphasized. PMID- 12894898 TI - A correlative study on serum Lp(a) in patients of cerebral infarction with that in their children. PMID- 12894899 TI - Changes of nailfold microcirculation in patients of type II diabetes mellitus with diabetic retinopathy. AB - OBJECTIVE: To study the changes of microcirculation in patients with diabetic retinopathy (DR). METHODS: Examination were performed in 153 cases of type I diabetes mellitus, among them, 72 cases were male, 81 cases were female, mean age 57.0+/-10.0 years, mean disease course 8.2+/-7.5 years. All cases were examined fundi by ophthalmologist, urinary albumin excretion rate (UAE) in 24 hours was measured by radioimmunoassay. Moreover, we examined the blood glucose, blood pressure, blood viscosity and observed the changes of naifold microcirculation. RESULTS: It was found that there were more evident disturbance of microcirculation, markedly slowed velocity of blood flow (P<0.05), significantly increased aggregation of blood cells (P<0.05) and exudation around the loop (P<0.05) in the group with DR, compared with the group without DR. CONCLUSION: It was more evident disturbance of nailfold microcirculation in patients with diabetic retinopathy. PMID- 12894900 TI - A study on PCR for detecting infection with M. leprae. AB - OBJECTIVE: So far, it has not been established a satisfactory method for early diagnosis and studying on epidemiology for leprosy, we want to develop a molecular biological method for solving this point. MATERIALS AND METHODS: Based on the M. leprae gene coding groEL, 65 kD and 16S rRNA, three polymerase chain reactions were developed by using Plikaytis', Woods' and Pattyn's procedures. It was optimized that the experimental parameters for each PCR, and a comparative study on practivity among three PCRs was also conducted for practical purpose. RESULTS AND CONCLUSION: For detecting infection with M. leprae, all of PCRs established by us were highly sensitive and specific, but for practical purpose, the Woods' PCR optimized by us ought to be chosen firstly. PMID- 12894901 TI - Experimental study on the segmental perfusion and preservation of spleen. AB - OBJECTIVE: Spleen transplanlation has developed to be an effective strategy for hemophilia A. But it has not been reported up to date that which kind of established solutions is most suitable for perfusion and preservation of spleen. This study aimed to establish some experiences with the comparison among Hartmann's solution, Collins' solution and WMO-I solution, in order to instruct the clinical spleen transplantation. METHODS: After the splenic artery and vein were dissociated clearly, three kinds of perfusion solutions began to perfuse the corresponding segments of spleen with a randomized sequence. When the efferent fluids from the splenic vein became clear, the perfused spleen segments were preserved for different durations with the same perfusing solution to calculate the survival rate of splencocyte (SRS) and were examined with light and electron microscopy. RESULTS: Among the three solutions, SRS with WMO-I solution was significantly higher than those of the other two (P<0.001). The perfused spleen with WMO-I solution showed the slightest morphological changes and a significant longer preservation duration than those with the other two (P<0.05 or P<0.01). CONCLUSION: Among the three solutions, WMO-I solution was most suitable for perfusion and preservation of spleen. PMID- 12894902 TI - Diagnosis and treatment of the Mirizzi syndrome. AB - OBJECTIVES: The aim of this paper is to describe the clinical characteristics, diagnostic procedure and operative management of Mirrizi syndrome. METHODS: Sixteen cases of Mirrizi syndrome were selected and reviewed from 1987 to 1997. RESULTS: In the 16 cases, 6 cases were male, 10 cases were female, the average age was 62.7 years old. Ten cases were diagnosed to be Mirrizi syndrome preoperatively (62.5%); 3 cases were considered to bile duct tumor, the other 3 cases were emergency, they were confirmed the diagnosis after the operation. CONCLUSIONS: Ultrasound is recommended as the first choice of screening method, while ERCP may confirm the diagnosis. Surgical approach is considered to be the choice and technical procedures are suggested to prevent intraoperative injury and to repair defects of the common bile duct. PMID- 12894903 TI - Apolipoprotein(a) and cortical cerebral infarction. AB - OBJECTIVE: The present study aimed to elucidating the correlation of apolipoprotein(a) [apo(a)] with cerebral infarction at the levels of molecule and protein. METHODS: The serum Lp(a) level, by means of ELISA, the polymorphism of apo(a) protein, by SDS-PAGE/Western blotting combined with silver staining assays, and the sequence polymorphisms in 5' control region, the first exon and intron of apo(a) gene, by PCR-SSCP/AFLP assays, were detected in 85 healthy controls and 12 cases of cortical cerebral infarction. RESULTS: The serum Lp(a) level was markedly higher in the patients (152.59+/-3.41 mg/L) than that in the controls (56.21+/-3.67 mg/L) (t=4, 15, P<0. 001), even between the subjects with the same apo(a) phenotype. The frequency of low molecular weight phenotype was significantly higher in the patients than that in the controls (0.5238 vs. 0.2941). There were 2 sites of sequence variance in the 5' control region of apo(a) gene in our studied population, which were of significant difference between patients and controls, and were related to the variation of serum Lp(a) level. CONCLUSION: Our study found that the low molecular weight phenotype of apo(a) was closely associated with cerebral infarction, suggested that the variation of serum Lp(a) level be determined by not only the size of apo(a) gene but also its sequence, which indicated that both the size and sequence of apo(a) are associated with the susceptibility to cerebral infarction. PMID- 12894904 TI - Morphological changes of rats muscles at various postmortem intervals by scanning electron microscopy. AB - The aim of this study was to observe the morphological changes of muscle in the process of rigor mortis. The quadriceps of 40 rats at various postmortem intervals were observed under the scanning electron microscope (SEM) and the light microscope by phosphotungstic acid-haematoxylin (PTAH) stain. The results showed that the striations of muscle were blurred within 4 h, but they became apparent from 6 h to 24 h after death. The authors suggest that this phenomenon be associated with the increased resistance of muscle against the postmortal changes. The observations by scanning electron microscopy and light microscopy have revealed that the muscles do contract in the process of rigor mortis because the distance between two Z lines shortens and the 1 band narrows, compared with those in anaesthetised animals. The basic biochemical process for the formation of rigor mortis is the same as that of muscle contraction except that the former happens postmortem and the latter antemortem. PMID- 12894905 TI - Effects of Tryptergium wilfordii polyglyco-sidium on the interleukin 1 levels in aqueous humor after intraocular lens implantation. PMID- 12894907 TI - 2002 Achievement Award of the IEEE Ultrasonics, Ferroelectrics, and Frequency Control Society. PMID- 12894906 TI - 2002 Rayleigh Award of the IEEE Ultrasonics, Ferroelectrics and Frequency Control Society Ultrasonics Committee. PMID- 12894908 TI - 2002 Distinguised Service Award of the IEEE Ultrasonics, Ferroelectrics and Frequency Control Society Ultrasonics Committee. PMID- 12894909 TI - 2001 Outstanding Paper Award of the IEEE Ultrasonics, Ferroelectrics, and Frequency Control Society. PMID- 12894910 TI - Two-dimensional equations for electroelastic plates with relatively large shear deformations. AB - A set of two-dimensional, nonlinear equations for electroelastic plates in moderately large thickness-shear deformations is obtained from the variational formulation of the three-dimensional equations of nonlinear electroelasticity by expanding the mechanical displacement vector and the electric potential into power series in the plate thickness coordinate. As an example, the equations are used to study nonlinear thickness-shear vibrations of a quartz plate driven by an electrical voltage. Nonlinear electrical current amplitude-frequency behavior near resonance is obtained. The equations and results are useful in the study and design of pieszoelectric crystal resonators and the measurement of nonlinear material constants of electroelastic materials. PMID- 12894911 TI - Transmitting electric energy through a metal wall by acoustic waves using piezoelectric transducers. AB - The feasibility of transmitting electric energy through a metal wall by propagating acoustic waves using piezoelectric transducers is examined by studying the efficiency of power transmission and its dependence upon the relevant system parameters for a simplified system consisting of an elastic plate sandwiched by two piezoelectric layers. One of these layers models the driving transducer for generating acoustic wave, and the other layer models the receiving transducer for converting the acoustic energy into electric energy to power a load circuit. The output voltage, the output power, and the efficiency of this system are expressed as explicit functions of the system parameters. A numerical example is included to illustrate the dependence of the system performance upon the physical and geometrical parameters. PMID- 12894912 TI - A 1.6-mm, metal tube ultrasonic motor. AB - A miniaturized metal tube ultrasonic motor, the dimensions of which are 1.6 mm in diameter and 6 mm in length, was developed. Two flattened surfaces with 90 degrees were ground on the outer surface of the stator. Two PZT-based piezoelectric ceramics were bonded onto these flat surfaces. The asymmetrical surface of the stator developed the split of the two degenerated orthogonal bending modes, resulting in a wobble motion. The working frequency of the 1.6-mm motor with 6 mm in length was 130 kHz. A torque of 0.5 mNm was reached at a maximum power of 45 mW with a speed of 45 rad/sec. The maximum efficiency was 16%. PMID- 12894913 TI - AO-spectrum analyzer implementation into CDMA-telecommunications with enhanced SIR-factors. AB - The devices for real-time narrowband interference measurements within the code division multiple access (CDMA) cellular communication system on the base of the acousto-optic spectrum analyzer (AOSA) with satisfactory performances can be designed. The overlay model giving significant advantages in the interference immunity for simultaneous operation of the wideband (CDMA) and the narrowband time division multiple access (TDMA) cellular systems is proposed. PMID- 12894914 TI - Autofocusing in medical ultrasound: the scaled covariance matrix algorithm. AB - This work develops a class of ultrasound phase aberration correction/autofocusing algorithms that are based upon the properties of the covariance matrix of the channel signals for time-delay focused resolution/speckle cells. The scaled covariance matrix (SCM) algorithms are designed to blindly estimate and correct focusing timing errors due to thin layers of unanticipated fatty tissue located in the near field of the transducer array. An important aspect of the algorithm is that the scaling of the covariance matrix elements fundamentally establishes a channel independent phase reference relative to which the aberrant channel phases are estimated. The model development involved the combination of a rigorous mathematical analysis of the scattering of ultrasound in random scattering media and extensive statistical simulation studies with phase aberrations imposed upon both the transmit and received channel signals. Under the assumption of a near field aberration model, the statistical simulation analyses showed that the SCM algorithms in simulation are capable of accurately estimating relative time delay channel errors with rms timing errors up to approximately 62 ns, with interchannel correlation lengths as short as 1.4 mm. PMID- 12894915 TI - Coded excitation with spectrum inversion (CEXSI) for ultrasound array imaging. AB - In this paper, a scheme called coded excitation with spectrum inversion (CEXSI) is presented. An established optimal binary code whose spectrum has no nulls and possesses the least variation is encoded as a burst for transmission. Using this optimal code, the decoding filter can be derived directly from its inverse spectrum. Various transmission techniques can be used to improve energy coupling within the system pass-band. We demonstrate its potential to achieve excellent decoding with very low (< 80 dB) side-lobes. For a 2.6 micros code, an array element with a center frequency of 10 MHz and fractional bandwidth of 38%, range side-lobes of about 40 dB have been achieved experimentally with little compromise in range resolution. The signal-to-noise ratio (SNR) improvement also has been characterized at about 14 dB. Along with simulations and experimental data, we present a formulation of the scheme, according to which CEXSI can be extended to improve SNR in sparse array imaging in general. PMID- 12894916 TI - Adaptive clutter rejection filtering in ultrasonic strain-flow imaging. AB - This paper introduces strain-flow imaging as a potential new technique for investigating vascular dynamics and tumor biology. The deformation of tissues surrounding pulsatile vessels and the velocity of fluid in the vessel are estimated from the same data set. The success of the approach depends on the performance of a digital filter that must separate echo signal components caused by flow from tissue motion components that vary spatially and temporally. Eigenfilters, which are an important tool for naturally separating signal components adaptively throughout the image, perform very well for this task. The method is examined using two tissue-mimicking flow phantoms that provide stationary and moving clutter associated with pulsatile flow. PMID- 12894917 TI - Sample volume misregistration in linear array-based dual beam Doppler ultrasound systems. AB - Large velocity estimation errors can occur in dual beam Doppler ultrasound velocity measurement systems when there is left/right sample volume misregistration, particularly when the interbeam angle is small. Such misregistration will occur when there is tissue inhomogeneity. This is investigated for a typical type of inhomogeneity--a layer of fat--by calculating the amount of both angle and translation misregistration occurring in such a system realized using a single linear array transducer. The complex sample volume sensitivity is calculated using a modified time domain approach, combining the spatial impulse response method with ray tracing. The effects on these misregistrations of altering the aperture sizes and their relative positions on the array is then investigated to derive an improved aperture configuration for dual beam velocity estimation. Arrangements with transmit apertures wider than the receive apertures are shown to be preferable in this context. PMID- 12894918 TI - In-vivo synthetic aperture flow imaging in medical ultrasound. AB - A new method for acquiring flow images using synthetic aperture techniques in medical ultrasound is presented. The new approach makes it possible to have a continuous acquisition of flow data throughout the whole image simultaneously, and this can significantly improve blood velocity estimation. Any type of filter can be used for discrimination between tissue and blood flow without initialization, and the number of lines used for velocity estimation is limited only by the nonstationarity of the flow. The new approach is investigated through both simulations and measurements. A flow rig is used for generating a parabolic laminar flow, and a research scanner is used for acquiring RF data from individual transducer elements. A reference profile is calculated from a mass flow meter. The parabolic velocity profile is estimated using the new approach with a relative standard deviation of 2.2% and a mean relative bias of 3.4% using 24 pulse emissions at a flow angle of 45 degrees. The 24 emissions can be used for making a full-color flow map image. An in-vivo image of flow in the carotid artery for a 29-year-old male also is presented. The full image is acquired using 24 emissions. PMID- 12894919 TI - Directional velocity estimation using focusing along the flow direction. I: Theory and simulation. AB - A new method for directional velocity estimation is presented. The method uses beamformation along the flow direction to generate data in which the correct velocity magnitude can be directly estimated from the shift in position of the received consecutive signals. The shift is found by cross-correlating the beamformed lines. The approach can find the velocity in any direction, including transverse to the traditionally emitted ultrasound beam. The velocity estimation is studied through extensive simulations using Field II. A 128-element, 7-MHz linear array is used. A parabolic velocity profile with a peak velocity of 0.5 m/s is simulated for different beam-to-flow angles and for different emit foci. At 45 degrees the relative standard deviation over the profile is 1.6% for a transmit focus at 40 mm. At 90 degrees the approach gave a relative standard deviation of 6.6% with a transmit focus of 80 mm, when using 8 pulse-echo lines and stationary echo canceling. Pulsatile flow in the femoral artery was also simulated using Womersley's flow model. A purely transverse flow profile could be obtained with a relative standard deviation of less than 10% over the whole cardiac cycle using 8 pulse emissions for each imaging direction, which is sufficient to show clinically relevant transverse color flow images. PMID- 12894920 TI - Directional velocity estimation using focusing along the flow direction. II: Experimental investigation. AB - A new method for directional velocity estimation is investigated through a number of flow rig measurements. The method uses beamformation along the flow direction to generate data, where the correct velocity magnitude can directly be estimated from the shift in position of the received consecutive signals. The shift is found by cross-correlating the beamformed lines. The approach can find the velocity in any direction, including transverse to the traditionally emitted ultrasound beam. The method is investigated using a flow rig with a peak velocity of 0.15 m/s. A 7-MHz linear array transducer is used together with a dedicated sampling system to acquire signals from 64 transducer elements simultaneously. A technique for obtaining 128-element data using multiplexing is also presented. The data is beamformed off-line on a PC. A relative standard deviation of 1.4% can be obtained for a beam-to-flow angle of 45 degrees and 4.3% at 90 degrees. Color flow images are displayed showing that the correct velocity magnitude can be obtained with the method for beam-to-flow angles of 60 and 90 degrees with an accuracy of 3 to 4%. PMID- 12894921 TI - Multifrequency ultrasound transducers for conformal interstitial thermal therapy. AB - Control over the pattern of thermal damage generated by interstitial ultrasound heating applicators can be enhanced by changing the ultrasound frequency during heating. The ability to change transmission frequency from a single transducer through the use of high impedance front layers was investigated in this study. The transmission spectrum of multifrequency transducers was calculated using the KLM equivalent circuit model and verified with experimental measurements on prototype transducers. The addition of a quarter-wavelength thick PZT (unpoled) front layer enabled the transmission of ultrasound at two discrete frequencies, 4.7 and 9.7 MHz, from a transducer with an original resonant frequency of 8.4 MHz. Three frequency transmission at 3.3, 8.4, and 10.8 MHz was possible for a transducer with a half-wavelength thick front layer. Calculations of the predicted thermal lesion size at each transmission frequency indicated that the depth of thermal lesion could be varied by a factor of 1.6 for the quarter wavelength front layer. Heating experiments performed in excised liver tissue with a dual-frequency applicator confirmed this ability to control the shape of thermal lesions during heating to generate a desired geometry. Practical interstitial designs that enable the generation of shaped thermal lesions are feasible. PMID- 12894922 TI - A complex virtual source approach for calculating the diffraction beam field generated by a rectangular planar source. AB - In this paper, a complex virtual source approach for calculating the ultrasound field generated by a rectangular planar source is presented. Instead of using a real rectangular plane source, the equivalent sources that have complex amplitudes in complex space are used to compute the sound field distribution. The parabolic equation first is solved in the kappa-space domain by applying Fourier transform. The kappa-space domain source is then expressed as a set of Gaussian functions, and the related coefficients is determined by the optimization method. The analytic solution then is derived, and the effect of the parameters on the calculation accuracy is discussed. The comparison between the proposed fast numerical scheme and previous methods (Fresnel integral and Ocheltree's method) and are given in an example. The numerical results reveal that the computation time in obtaining accurate calculations is greatly reduced by using the proposed method. PMID- 12894923 TI - An acoustic proximity ranging system for monitoring the cavity thickness. AB - To control high speed underwater vehicles, a proximity ranging system is needed to monitor the cavity thickness. In this paper, we study a time-of-flight (TOF) principle based acoustic proximity ranging system. By taking into account the acoustically hard boundary at the air-water interface, we first present a two stage computationally efficient time delay estimation algorithm, referred to as the PEARS (Parameter Estimation for Acoustic Ranging Systems) algorithm, which is applicable to arbitrary transmitted waveforms. Numerical results based on a simulated waveform demonstrate that the PEARS estimates can approach the Cramer Rao bound as the signal-to-noise ratio increases. We then present experiments performed by using commercially available acoustic transducers to further verify our method. To update TOF estimates quickly, a specially designed continuous wave (CW) is applied to the transducer. Experimental results show that PEARS can achieve high measurement accuracy for ranging distances less than 100 mm with an achievable parameter update rate of approximately 1.5 kHz. PMID- 12894924 TI - Robust control of linear ceramic motor drive with LLCC resonant technique. AB - This study presents a robust control system for a linear ceramic motor (LCM) that is driven by a high-frequency voltage source inverter using two-inductance two capacitance (LLCC) resonant technique. The structure and driving principle of the LCM are introduced. Because the dynamic characteristics and motor parameters of the LCM are nonlinear and time varying, a robust control system is designed based on the hypothetical dynamic model to achieve high-precision position control. The presentation of robust control for the LCM drive system is divided into three parts, which comprise state feedback controller, feed-forward controller, and uncertainty controller. The adaptation laws of control gains in the robust control system are derived in the sense of Lyapunov stability theorem such that the stability of the control system can be guaranteed. It not only has the learning ability similar to intelligent control, but also its control framework is more simple than intelligent control. With the proposed robust control system, the controlled LCM drive possesses the advantages of good tracking control performance and robustness to uncertainties. The effectiveness of the proposed robust control system is verified by experimental results in the presence of uncertainties. In addition, the advantages of the proposed control system are indicated in comparison with the traditional integral-proportional (IP) position control system. PMID- 12894925 TI - Electric poling and electromechanical characterization of 0.1-mm-thick sensor films and 0.2-mm-thick cable layers from piezoelectric poly(vinylidene fluoride trifluoroethylene). AB - Piezoelectric polymers have been proposed for many sensor and actuator applications. Among these, piezoelectric polymer films with thicknesses between several tens and a few hundreds of micrometers as well as coaxial cables with piezoelectric polymer layers are highly suitable and attractive for the detection of mechanical loads. In addition to good piezoelectric properties, materials for such sensors should have high mechanical strength. Therefore, the most common materials are nonporous piezoelectric polymers, such as polyvinylidene fluoride (PVDF) or its copolymer with trifluoroethylene (P(VDF-TrFE)). Here, P(VDF-TrFE) polymer films as well as the operating principle and the geometry of piezoelectric polymer cables are described. As active piezoelectric cable layer, P(VDF-TrFE) (76/24) was employed. After electrical poling with one or more point to-cable corona discharges, the polarization in the P(VDF-TrFE) layer was investigated. Poling parameters, such as electric field and poling time, were varied. The resulting polarization was characterized with measurements of polarization profiles across the film or the cable-layer thickness as well as with the determination of other electromechanical quantities. The optimized poling process yields good piezoelectric properties in the piezoelectric polymer layers as well as useful sensor properties of the piezoelectric polymer cable, which are assessed and discussed. PMID- 12894926 TI - In-air and underwater performance and finite element analysis of a flextensional device having electrostrictive poly(vinylidene fluoride-trifluoroethylene) polymers as the active driving element. AB - A flextensional transducer, in which the electrostrictive poly(vinylidene fluoride-trifluoroethylene) [P(VDF-TrFE)] copolymer was used as the active driving element, was fabricated and characterized. The results show that transducers of several millimeters thick can produce an axial displacement of more than 1 mm in air along the thickness direction, and a transmitting voltage response of 123 dB re 1 microPa/V at 1 m in water at frequencies of several kilohertz. A finite element code (ANSYS, Inc., Canonsburg, PA) was used to model the in-air and underwater responses of the flextensional transducer over a broad frequency range. The calculated resonance frequencies and transmitting voltage response spectra show good agreement with the experimental data. In addition, the performance of both the in-air actuator and underwater transducer was analyzed for different design parameters of the flextensional structure. These results show that the performance of the flextensional transducer could be tailored readily by adjusting the parameters of the flextensional metal shell. PMID- 12894927 TI - Efficacy or effectiveness: which comes first, the cure or the treatment? PMID- 12894928 TI - Efficacy vs. effectiveness research in psychotherapy: implications for clinical hypnosis. AB - Empirically supported therapy (EST) has become a major focus and trend for mental health practice. When hypnosis is involved, this may mean satisfying a standard that is entirely too narrow in its emphasis. In this article "efficacy"-based research in clinical practice is contrasted with "effectiveness" -focused research, and they are discussed from the perspective of hypnosis. When clinicians can consider trans-theoretical factors as well as those that are treatment-enhancing, possibilities for improved treatment outcome increase. The "effectiveness" perspective also serves as a counter point for hypnosis in contrast with the dubious efficacy-based gold standard currently proposed for therapy in general, and hypnosis in particular. PMID- 12894929 TI - A project to change the attitudes, beliefs and practices of health professionals concerning hypnosis. AB - This study describes an educational project designed to change the beliefs, attitudes and practices of health care professionals concerning hypnosis and hypnotherapy. The purpose of the study was to investigate whether an educational offering that dispelled myths and misconceptions of hypnosis, offered a historical perspective, discussed how hypnosis works therapeutically, and elucidated the many medical uses of hypnosis, could change the attitudes, practices and beliefs of health care providers. Workshops were given to health care professionals at a variety of venues. Three questionnaires were used to collect data from study subjects before, immediately after and three months following the educational intervention. The results indicated that it is possible with a 90 to 180 minute lecture on hypnosis to create changes in beliefs, attitudes and practices. When health professionals have accurate information on hypnosis that is substantiated by scientific research, they are more likely to believe that hypnosis has a place in traditional mainstream medicine and to suggest self-regulatory strategies and hypnosis to their patients. An educational presentation on hypnosis can encourage health care professionals to learn more about this modality and to seek training in hypnosis. PMID- 12894930 TI - The hypnotic belay in alpine mountaineering: the use of self-hypnosis for the resolution of sports injuries and for performance enhancement. AB - The author, an experienced alpine mountaineer, sustained several traumatic climbing injuries over a two-year period. This article describes her multiple uses of self-hypnosis to deal with several challenges related to her returning to successful mountain climbing. She used self-hypnosis for physical healing and to enhance her motivation to resume climbing. While training for her next expedition, she successfully utilized self-hypnotic techniques to deal with acute stress and later post-traumatic symptoms that had emerged related to her climbing injuries. She describes her use of hypnotic ego-strengthening, mental rehearsal, age progression, and "Inner Strength" as well as active-alert trance states. Her successful summitting of Ecuador's Cotopaxi at 19,380 feet was facilitated by "The Hypnotic Belay" which permitted her to secure herself by self-hypnosis in addition to the rope used to secure climbers. In 1994, the author returned to the Cascade Mountains where she had been injured three years earlier and reached the summit of Mount Shuksan. This time she was secured by "The Hypnotic Belay". PMID- 12894931 TI - Perspective: sexual conflict and sexual selection: chasing away paradigm shifts. AB - Traditional models of sexual selection propose that partner choice increases both average male and average female fitness in a population. Recent theoretical and empirical work, however, has stressed that sexual conflict may be a potent broker of sexual selection. When the fitness interests of males and females diverge, a reproductive strategy that increases the fitness of one sex may decrease the fitness of the other sex. The chase-away hypothesis proposes that sexual conflict promotes sexually antagonistic, rather than mutualistic, coevolution, whereby manipulative reproductive strategies in one sex are counteracted by the evolution of resistance to such strategies in the other sex. In this paper, we consider the criteria necessary to demonstrate the chase-away hypothesis. Specifically, we review sexual conflict with particular emphasis on the chase-away hypothesis; discuss the problems associated with testing the predictions of the chase-away hypothesis and the extent to which these predictions and the predictions of traditional models of sexual selection are mutually exclusive; discuss misconceptions and mismeasures of sexual conflict; and suggest an alternative approach to demonstrate sexual conflict, measure the intensity of sexually antagonistic selection in a population, and elucidate the coevolutionary trajectories of the sexes. PMID- 12894932 TI - Detecting correlation between characters in a comparative analysis with uncertain phylogeny. AB - The importance of accommodating the phylogenetic history of a group when performing a comparative analysis is now widely recognized. The typical approaches either assume the tree is known without error, or they base inferences on a collection of well-supported trees or on a collection of trees generated under a stochastic model of cladogenesis. However, these approaches do not adequately account for the uncertainty of phylogenetic trees in a comparative analysis, especially when data relevant to the phylogeny of a group are available. Here, we develop a method for performing comparative analyses that is based on an extension of Felsenstein's independent contrasts method. Uncertainties in the phylogeny, branch lengths, and other parameters are accommodated by averaging over all possible trees, weighting each by the probability that the tree is correct. We do this in a Bayesian framework and use Markov chain Monte Carlo to perform the high-dimensional summations and integrations required by the analysis. We illustrate the method using comparative characters sampled from Anolis lizards. PMID- 12894933 TI - Visual background complexity facilitates the evolution of camouflage. AB - Cryptic animal coloration or camouflage is an adaptation that decreases the risk of detection. The study of the evolution of camouflage has strongly emphasized the minimization of visual information that predators receive from prey, by means of background matching. However, the evolutionary effects of information processing after its reception have been virtually ignored. I constructed a model that employs an artificial neural network and simulates the evolution of prey coloration in a visually complex and simple habitat. The model suggests: (1) the difficulty of a detection task is related to the visual complexity of the habitat; (2) it is easier to decrease the risk of detection by the means of camouflage in a visually complex habitat; (3) selection on camouflage can exploit limitations in predators information processing; and (4) there are shortcomings in using the degree of background matching as the measure of camouflage. PMID- 12894934 TI - Convergence and coevolution in a mutualism: evidence from a molecular phylogeny of Ficus. AB - The interaction between Ficus (Moraceae) and their pollinating wasps (Chalcidoidea: Agaonidae; more than 700 species-specific couples) is one of the most specialized mutualisms found in nature. Both partners of this interaction show extensive variation in their respective biology. Here we investigate Ficus life-history trait evolution and fig/fig wasp coadaptation in the context of a well-resolved molecular phylogeny. Mapping out variations in Ficus life-history traits on an independently derived phylogeny constructed from ribosomal DNA sequences (external and internal transcribed spacer) reveals several parallel transitions in Ficus growth habit and breeding system. Convergent trait evolution might explain the discrepancies between morphological analyses and our molecular reconstruction of the genus. Morphological characters probably correlate with growth habit and breeding system and could therefore be subject to convergent evolution. Furthermore, we reconstruct the evolution of Ficus inflorescence characters that are considered adaptations to the pollinators. Our phylogeny reveals convergences in ostiole shape, stigma morphology, and stamen:ovule ratio. Statistical tests taking into account the phylogenetic relationship of the species show that transitions in ostiole shape are correlated with variation in wasp pollinator head shape, and evolutionary changes in stigma morphology and stamen:ovule ratio correlate with changes in the pollination behavior of the associated wasp. These correlations provide evidence for reciprocal adaptations of morphological characters between these mutualistic partners that have interacted over a long evolutionary time. In light of previous ecological studies on mutualism, we discuss the adaptive significance of these correlations and what they can tell us about the coevolutionary process occurring between figs and their pollinators. PMID- 12894935 TI - Evolutionary genetics of resistance and tolerance to natural herbivory in Arabidopsis thaliana. AB - Resistance and tolerance are widely viewed as two alternative adaptive responses to herbivory. However, the traits underlying resistance and tolerance remain largely unknown, as does the genetic architecture of herbivory responses and the prevalence of genetic trade-offs. To address these issues, we measured resistance and tolerance to natural apical meristem damage (AMD) by rabbits in a large field experiment with recombinant inbred lines (RILs) of Arabidopsis thaliana (developed from a cross between the Columbia x Landsberg erecta ecotypes). We also measured phenological and morphological traits hypothesized to underlie resistance and tolerance to AMD. Recombinant inbred lines differed significantly in resistance (the proportion of replicates within an RIL that resisted herbivory), and early flowering plants with tall apical inflorescences were more likely to experience damage. Tolerance (the difference in fitness between the damaged and undamaged states), also differed significantly among RILs, with some lines overcompensating for damage and producing more fruit in the damaged than undamaged state. Plastic increases in basal branch number, basal branch height, and senescence date in response to damage were all associated with greater tolerance. There was no evidence for a genetic trade-off between resistance and tolerance, an observation consistent with the underlying differences in associated morphological and phenological characters. Selection gradient analysis detected no evidence for direct selection on either resistance or tolerance in this experiment. However, a statistical model indicates that the pattern of selection on resistance depends strongly on the mean level of tolerance, and selection on tolerance depends strongly on the mean level of resistance. These observations are consistent with the hypothesis that selection may act to maintain resistance and tolerance at intermediate levels in spatially or temporally varying environments or those with varying herbivore populations. PMID- 12894936 TI - Dramatic phenotypic plasticity in barnacle legs (Balanus glandula Darwin): magnitude, age dependence, and speed of response. AB - The precise dependence of barnacle leg form on flow suggests the wave-swept environment imposes strong selection on suspension feeding limbs. I conducted three experiments to determine the mechanism, age dependence, and response time of cirrus variation in the acorn barnacle Balanus glandula. (1) To test whether cirrus variation arises via genetic or environmental mechanisms, I transplanted juvenile barnacles from one wave-exposed and one protected population into high and low flow conditions. Both populations exhibited similar abilities to modify cirri in response to experimental velocities: transplanted barnacles grew legs up to 84% longer in low flow. A small (up to 24%), but significant difference between source populations suggested slight genetic divergence in leg form. (2) Because flow is heterogeneous over space and time, I tested whether cirrus plasticity was limited to juveniles by transplanting both juveniles and adults from exposed and protected shores into quiet water. Remarkably, both juveniles and adults from the wave-exposed population produced legs over 100% longer than the original population, whereas protected barnacles remained unchanged. (3) A third transplant of adults into quiet water demonstrated that wave-exposed B. glandula modified cirrus form very quickly-within 18 days, or one to two molts. Results from these experiments suggest that variation in cirrus form is largely environmentally induced, but genetic differences may account for some variation observed among field populations; spatial and temporal flow heterogeneity appear to have selected for extreme flexibility of feeding form throughout a barnacle's life; and flow heterogeneity in the wave-swept environment appears to have selected for rapid ecophenotypic responses in the form of feeding structures. PMID- 12894937 TI - Phylogenetic evidence for hybrid origins of asexual lineages in an aphid species. AB - Understanding the mode of origin of asexuality is central to ongoing debates concerning the evolution and maintenance of sexual reproduction in eukaryotes. This is because it has profound consequences for patterns of genetic diversity and ecological adaptability of asexual lineages, hence on the outcome of competition with sexual relatives both in short and longer terms. Among the possible routes to asexuality, hybridization is a very common mechanism in animals and plants. Aphids present frequent transitions from their ancestral reproductive mode (cyclical parthenogenesis) to permanent asexuality, but the mode of origin of asexual lineages is generally not known because it has never been thoroughly investigated with appropriate molecular tools. Rhopalosiphum padi is an aphid species with coexisting sexual (cyclically parthenogenetic) and asexual (obligately parthenogenetic) lineages that are genetically distinct. Previous studies have shown that asexual lineages of R. padi are heterozygous at most nuclear loci, suggesting either that they have undergone long-term asexuality (under which heterozygosity tends to increase) or that they have hybrid origins. To discriminate between these alternatives, we conducted an extensive molecular survey combining the sequence analysis of alleles of two nuclear DNA markers and mitochondrial DNA haplotypes in sexual and asexual lineages of R. padi. Both nuclear and cytoplasmic markers clearly showed that many asexual lineages have hybrid origins, the first such demonstration in aphids. Our results also indicated that asexuals result from multiple events of hybridization between R. padi and an unknown sibling species, and are of recent origin (contradicting previous estimates that asexual R. padi lineages were of moderate longevity). This study constitutes another example that putatively ancient asexual lineages are actually of much more recent origin than previously thought. It also presents a robust approach for testing whether hybrid origin of asexuality is also a common phenomenon in aphids. PMID- 12894938 TI - Genetic variance and phenotypic plasticity in a component of female mate choice in an ultrasonic moth. AB - Female response to male advertisement signals in lesser waxmoths showed substantial genetic variation, phenotypic plasticity across rearing environments, and genotype-by-environment interactions resulting in crossing reaction norms. These results represent two previously underemphasized means by which genetic variation may be maintained in sexually selected traits: genetic variation in female response to male traits, and variation in the selection acting on both males and females. Genotype-by-environment interactions and reaction norms that cross indicate that divergent selection may act on male and female sexual traits if the level of environmental change is high. The processes that contribute to the maintenance of genetic variation may thus also contribute to population differentiation. PMID- 12894939 TI - The influence of variable rates of inbreeding on fitness, environmental responsiveness, and evolutionary potential. AB - We manipulated experimental populations of the housefly (Musca domestica L.) under three inbreeding schemes (fast, slow, and punctuated) to partition out the influences of different means and variances in the rate of inbreeding, per generation, while controlling for the final level of inbreeding as a constant. One treatment used constant fast inbreeding (11% per generation; Ne = 4 for 4 generations), for a comparison to one that was consistently slow (3% per generation; Ne = 16 for 14 generations). The third followed a model for serial founder-flush events. Each founder-flush episode involved a one-generation pulse of fast inbreeding (Ne = 4) followed by two generations of very low (or no) inbreeding, yielding high intergenerational variation (i.e., for an average inbreeding rate of 4% per generation). Allozyme assays showed that we achieved the intended final inbreeding coefficient of about 37%. All inbreeding schemes decreased fitness levels in terms of egg-to-adult viability, development time, and male mating success relative to the outbred control. The consistently fast inbreeding protocol had more pronounced reductions in fitness, relative to the other two inbreeding schemes. In comparison to the fast and punctuated regimes, the consistently slow protocol preserved evolutionary potential (as assayed by the genetic divergence of subpopulations exposed to different environments) in egg-to-adult viability, and (albeit anecdotally) reduced the extinction probabilities, especially in a novel environment. The punctuated treatment did not optimize the potential for purge as predicted, but instead reduced fitness, evolutionary potential, and environmental responsiveness (as measured by genotype by-environment interactions). This founder-flush treatment also had the highest extinction probabilities. Longer periods of population flush might be necessary to purge effectively in a punctuated scheme. We conclude that the rate of inbreeding, independent from the final level, can have important effects on population fitness, environmental responsiveness, and evolutionary potential. PMID- 12894940 TI - Low occurrence of gene transposition events during the evolution of the genus Drosophila. AB - The role played by gene transpositions during the evolution of eukaryotic genomes is still poorly understood and indeed has been analyzed in detail only in nematodes. In Drosophila, a limited number of transpositions have been detected by comparing the chromosomal location of genes between different species. The relative importance of gene transposition versus other types of chromosomal rearrangements, for example, inversions, has not yet been evaluated. Here, we use physical mapping to perform an extensive search for long-distance gene transpositions and assess their impact during the evolution of the Drosophila genome. We compare the relative order of 297 molecular markers that cover 60% of the euchromatic fraction of the genome between two related Drosophila species and conclude that the frequency of gene transpositions is very low, namely one order of magnitude lower than that of nematodes. In addition, gene transpositions seem to be events almost exclusively associated with genes of repetitive nature such as the Histone gene complex (HIS-C). PMID- 12894941 TI - Colony sex ratios vary with breeding system but not relatedness asymmetry in the facultatively polygynous ant Pheidole pallidula. AB - We investigated sex allocation in a Mediterranean population of the facultatively polygynous (multiple queen per colony) ant Pheidole pallidula. This species shows a strong split sex ratio, with most colonies producing almost exclusively a single-sex brood. Our genetic (microsatellite) analyses reveal that P. pallidula has an unusual breeding system, with colonies being headed by a single or a few unrelated queens. As expected in such a breeding system, our results show no variation in relatedness asymmetry between monogynous (single queen per colony) and polygynous colonies. Nevertheless, sex allocation was tightly associated with the breeding structure, with monogynous colonies producing a male-biased brood and polygynous colonies almost only females. In addition, sex allocation was closely correlated with colony total sexual productivity. Overall, our data show that when colonies become more productive (and presumably larger) they shift from monogyny to polygyny and from male production to female production, a pattern that has never been reported in social insects. PMID- 12894942 TI - Arboreallty and morphological evolution in ground beetles (Carabidae: Harpalinae): testing the taxon pulse model. AB - One-third to two-thirds of all tropical carabids, or ground beetles, are arboreal, and evolution of arboreality has been proposed to be a dead end in this group. Many arboreal carabids have unusual morphological features that have been proposed to be adaptations for life on vegetation, including large, hemispheric eyes; an elongated prothorax; long elytra; long legs; bilobed fourth tarsomeres; adhesive setae on tarsi; and pectinate claws. However, correlations between these features and arboreality have not been rigorously tested previously. I examined the evolution of arboreality and morphological features often associated with this habitat in a phylogenetic context. The number and rates of origins and losses of arboreality in carabids in the subfamily Harpalinae were inferred with parsimony and maximum-likelihood on a variety of phylogenetic hypotheses. Correlated evolution in arboreality and morphological characters was tested with concentrated changes tests, maximum-likelihood, and independent contrasts on optimal phylogenies. There is strong evidence that both arboreality and the morphological features examined originated multiple times and can be reversed, and in no case could the hypothesis of equal rates of gains and losses be rejected. Several features are associated with arboreality: adhesive setae on the tarsi, bilobed tarsomeres, and possibly pectinate claws and an elongated prothorax. Bulgy eyes, long legs, and long elytra were not correlated with arboreality and are probably not arboreal adaptations. The evolution of arboreal carabids has not been unidirectional. These beetles have experienced multiple gains and losses of arboreality and the morphological characters commonly associated with the arboreal habitat. The evolutionary process of unidirectional character change may not be as widespread as previously thought and reversal from specialized lifestyles or habitats may be common. PMID- 12894943 TI - Genetic variation in male effects on female reproduction and the genetic covariance between the sexes. AB - Males of many insect species increase the fecundity and/or egg size of their mates through the amount or composition of their nuptial gifts or ejaculate. The genetic bases of such male effects on fecundity or egg size are generally unknown, and thus their ability to evolve remains speculative. Likewise, the genetic relationship between male and female investment into reproduction in dioecious species, which is expected to be positive if effects on fecundity are controlled by at least some of the same genes in males and females, is also unknown. Males of the seed beetle Stator limbatus contribute large ejaculates to females during mating, and the amount of donated ejaculate is positively correlated with male body mass. Females mated to large males lay more eggs in their lifetime than females mated to small males. We describe an experiment in which we quantify genetic variation in the number of eggs sired by males (mated to a single female) and found that a significant proportion of the phenotypic variance in the number of eggs sired by males was explained by their genotype. Additionally, the number of eggs sired by a male was highly positively genetically correlated with his body mass. The between-sex genetic correlation, that is, the genetic correlation between the number of eggs sired by males and the number of eggs laid by females, was highly positive when eggs were laid on Acacia greggii seeds. This indicates that males that sire many eggs have sisters that lay many eggs. Thus, some of the genes that control male ejaculate size (or some other fecundity-enhancing factor) when expressed in males appear to control fecundity when expressed in females. We found no significant interaction between male and female genotype on fecundity. PMID- 12894944 TI - Do female spiders select heavier males for the genes for behavioral aggressiveness they offer their offspring? AB - We explore the hypothesis that females choose to mate with heavier males for the genes for behavioral aggressiveness they offer their offspring in the desert spider, Agelenopsis aperta. Behavioral aggressiveness is important to competition for limited resources in the field and is thus correlated with the mass spiders achieve. We established four crosses based on the body mass relationships of parents subjected to selection in their natural environment (female mass/male mass: HI/HI, HI/LO, LO/HI, and LO/LO) and reared the F1 offspring in a noncompetitive laboratory environment. Offspring size and mass at maturity were measured, life history parameters recorded, and behavioral aggressiveness scored in a series of tests. Significant familial effects were detected in all of these measures, but pertinent cross effects were observed only in the assays measuring behavioral aggressiveness. The results were summarized in terms of the fitness costs to HI females of mating with LO males (fewer female offspring of the more aggressive phenotypes) and the benefits to LO females of mating with HI males (fewer fearful offspring of both sexes). PMID- 12894945 TI - The dynamics of male brooding, mating patterns, and sex roles in pipefishes and seahorses (family Syngnathidae). AB - Modern theory predicts that relative parental investment of the sexes in their young is a key factor responsible for sexual selection. Seahorses and pipefishes (family Syngnathidae) are extraordinary among fishes in their remarkable adaptations for paternal care and frequent occurrences of sex-role reversals (i.e., female-female competition for mates), offering exceptional opportunities to test predictions of sexual selection theory. During mating, the female transfers eggs into or onto specialized egg-brooding structures that are located on either the male's abdomen or its tail, where they are osmoregulated, aerated, and nourished by specially adapted structures. All syngnathid males exhibit this form of parental care but the brooding structures vary, ranging from the simple ventral gluing areas of some pipefishes to the completely enclosed pouches found in seahorses. We present a molecular phylogeny that indicates that the diversification of pouch types is positively correlated with the major evolutionary radiation of the group, suggesting that this extreme development and diversification of paternal care may have been an important evolutionary innovation of the Syngnathidae. Based on recent studies that show that the complexity of brooding structures reflects the degree of paternal investment in several syngnathid species, we predicted sex-role reversals to be more common among species with more complex brooding structures. In contrast to this prediction, however, both parsimony- and likelihood-based reconstructions of the evolution of sex-role reversal in pipefishes and seahorses suggest multiple shifts in sex roles in the group, independent from the degree of brood pouch development. At the same time, our data demonstrate that sex-role reversal is positively associated with polygamous mating patterns, whereas most nonreversed species mate monogamously, suggesting that selection for polygamy or monogamy in pipefishes and seahorses may strongly influence sex roles in the wild. PMID- 12894946 TI - Peering through the looking glass at a sexual parasite: are Amazon mollies red queens? AB - The gynogenetic Amazon molly (Poecilia formosa) is a clonal, all-female lineage of livebearing fish that faces an unusual obstacle to evolutionary persistence. Sperm from heterospecific males (either sailfin, P. latipinna, or Atlantic, P. mexicana, mollies) is necessary to trigger embryogenesis. However, none of the male's genes are incorporated into the genome of the gynogenetic offspring. Some investigators have proposed that the evolution of male mate discrimination is a result of this cost, leading to a coevolutionary arms race between male avoidance of P. formosa and P. formosa attractiveness. Given that P. formosa successfully reproduces and has not yet gone extinct, it is clear there are mechanisms by which they attract the sexual attention of males. Although a Red Queen coevolutionary process in typical host/parasite systems has been shown to favor the persistence of sexual species, in this system an arms race has been invoked to explain the reverse. Here I present behavioral data supporting a more parsimonious scenario: that mechanisms of attraction in P. formosa are simply a consequence of its hybrid origin. Poecilia latipinna and P. mexicana males do not discriminate between gynogenetic P. formosa females and first generation sexual hybrid females, and females do not differ in agonistic behaviors associated with competition for mates. Both results contradict predictions from the Red Queen hypothesis. Therefore, coevolution is not necessary to explain the apparent evolutionary persistence of P. formosa. PMID- 12894947 TI - AFLP phylogeny of Mimulus section Erythranthe and the evolution of hummingbird pollination. AB - Species in Mimulus section Erythranthe (monkeyflowers) have become model systems for the study of the genetic basis of ecological adaptations. In this study, we pursued two goals. First, we reconstructed the phylogeny of species in Erythranthe using both DNA sequences from the ribosomal DNA ITS and ETS and AFLPs. Data from rDNA sequences support the monophyly of the section, including M. parishii, but provide little support for relationships within it. Analyses using AFLP data resulted in a well-supported hypothesis of relationships among all Erythranthe species. Our second goal was to reconstruct ancestral pollination syndromes and ancestral states of individual characters associated with hummingbird-pollinated flowers. Both parsimony and likelihood approaches indicate that hummingbird pollination evolved twice in Erythranthe from insect-pollinated ancestors. Our reconstruction of individual characters indicates that corolla color and some aspects of corolla shape change states at the same point on the phylogenetic tree as the switch to hummingbird pollination; however, a switch to secretion of high amounts of nectar does not. Floral trait transformation may have been more punctuational than gradual. PMID- 12894948 TI - Hybridization and postzygotic isolation patterns in pigeons and doves. AB - The study of the patterns of reproductive isolation in relation to species divergence is critical for the understanding of the process of speciation. Comparative analyses of this kind were previously conducted in Drosophila, Lepidoptera, frogs, ducks, and birds in general. In the present study, we used information from the literature to analyze hybrid inviability in relation to species divergence in pigeons and doves. Four main patterns arose from this analysis: (1) as in the other groups studied, F1 hybrid inviability gradually increases as species diverge, the time needed to reach total inviability being higher in birds than in the other groups; (2) as expected, the presence of geographic overlap does not influence the evolution of postzygotic isolation; (3) the percentage of unhatched eggs does not differ between hybrids of the first generation and the backcrosses, but it increases in the second hybrid generation; and (4) pigeons and doves follow Haldane's rule, as found in the other groups studied so far. The similarity between the results of this and previous studies contributes to the growing evidence suggesting that the patterns of the evolution of postzygotic isolation, and the process of speciation in general, are shared among animal groups. PMID- 12894949 TI - Paternal, maternal, and biparental molecular markers provide unique windows onto the evolutionary history of macaque monkeys. AB - We report the results of one of the first intrageneric analyses to simultaneously survey mitochondrial, Y-chromosomal, and autosomal loci from the same individuals representing the same taxa. Phylogenetic trees were constructed for each of these genetic systems from a pool of 63 macaques, representing all 19 recognized species in this genus, and eight outgroup taxa. The mitochondrial locus analyzed here (1.5 Kb) spans the 3' end of 12S rDNA, tRNA-VAL, and the 5' end of 16S rDNA; the Y chromosome dataset (3.1 Kb) consists of the genes SRY and TSPY; the two autosomal datasets include IRBP intron 3 (1.6 Kb) and the 5' half of C4 "long" intron 9 (3.3 Kb). A total of 1.35 million bases were read, revealing 682 variable sites within the genus Macaca. With regard to earlier unresolved issues of macaque evolution, a comparison of topologies reconstructed from each of the three genetic systems suggests: (1) four monophyletic species groups; (2) an initial bifurcation among Asian macaques between the silenus group progenitor and a M. fascicularis-like taxon, with the latter representing the probable common ancestor to all non-silenus group Asian macaques; (3) a possible hybrid origin of M. arctoides from proto-M. assamensis/thibetana and proto-M. fascicularis; and (4) contemporary introgression between M. mulatta and M. fascicularis in Indochina. Inferences 3 and 4 are of particular interest, because episodes of reticulate evolution often go undetected in analyses employing a single genetic system. Finally, divergence calculations suggest that, in female-philopatric taxa, mitochondrial bifurcations may typically predate Y-chromosomal divergences at the same node. PMID- 12894950 TI - Monkeys and toads define areas of endemism on Sulawesi. AB - Ecological or geological phenomena can impose limits on geographic diversification that cause biogeographical patterns of distantly related but sympatrically occurring taxa to be similar. Concordant patterns of diversity facilitate conservation management because strategic designation of protected areas can capture complementary rather than redundant components of variation. Here we demonstrate that on the biodiverse Indonesian island of Sulawesi, seemingly idiosyncratic distributions of diversity in endemic monkeys (Macaca species) and toads (Bufo celebensis) are actually virtually identical on a fine geographic scale. It appears that range fragmentation has generated seven multi taxon areas of genetic endemism, each of which should be targeted for conservation. Joint consideration of molecular phylogeography, morphology, and demography helps resolve apparent contradictions in paraphyletic macaque mitochondrial DNA and in undifferentiated toad morphology, and facilitates an understanding of biogeography and conservation genetics of Sulawesi fauna. PMID- 12894951 TI - Specific versus nonspecific immune defense in the bumblebee, Bombus terrestris L. AB - Hosts vary in both their strength of response to a general immunological insult and in their specific susceptibility to different parasite species or different strains of the same parasite. The variation in the general immune response is considered a result of the costs imposed by selection on defended individuals. The variation in the specific response may originate from variation in host and parasite genotypes and is a requirement for frequency-dependent selection. The relationship between these two fundamental aspects of defense has only rarely been studied. Using the bumblebee Bombus terrestris and its gut trypanosomal parasite Crithidia bombi we found that the host's specific response profile toward different strains correlates negatively with its level of response to a general insult. This is the opposite result one would expect if the level of general response were simply a measure of immunological quality (immunocompetence). Rather, it suggests that there is some form of a trade-off between these two fundamental aspects of the immune system. These results, therefore, shed an important light on the possible constraints that affect the evolution of the immune system and particularly the trade-off between different arms of the immune system. PMID- 12894952 TI - Is evolvability involved in the origin of modular variation? AB - Lipson et al. (2002) presented an elegant linear algebraic formalism to define and study the evolution of modularity in an artificial evolving system. They employed simulation data to support their suggestion that modularity arises spontaneously in temporally fluctuating systems in response to selection for enhanced evolvability. We show analytically and by simulation that their correlate of modularity is itself under selection and so is not a reliable indicator of selection for modularity per se. In addition, we question the relation between modularity and evolvability in their simulations, suggesting that this modularity cannot confer enhanced evolvability. PMID- 12894953 TI - Integrated health care must consider mental health to reduce rates of cardiovascular disease. PMID- 12894954 TI - Hypertension in Ghana: a cross-sectional community prevalence study in greater Accra. AB - OBJECTIVES: To determine the prevalence of hypertension, and the extent to which it is treated and controlled, among adult Ghanaians. DESIGN: 6300 adults, aged 25 years and older, were selected by random cluster sampling, using electoral enumeration areas and listings of adults. SETTING: Three communities in the greater Accra region of Ghana. SUBJECTS AND METHODS: A total of 4733 subjects (male to female ratio = 1:1.5) participated, representing a response rate of 75%. The analysis used the mean of 2 blood pressure readings, taken with a mercury sphygmomanometer after a 10-minute rest. Hypertension was defined as having blood pressure > or = 140/90 mm Hg, or currently undergoing anti-hypertensive treatment. RESULTS: The mean ages for males and females were 44.9 +/- 14.7 years, and 44.0 +/- 14.6 years, respectively. The crude prevalence of hypertension was 28.3%. The age-standardized prevalence, to the new standard world population, was 28.4%. Mean systolic and diastolic blood pressures increased with age. Of 1337 subjects with hypertension, 34% were aware of their condition, 18% were treated, and 4% were controlled (blood pressure < 140/90 mm Hg). CONCLUSION: Hypertension is a major public health problem, and is associated with relatively low levels of awareness, drug treatment, and blood pressure control. Population-based prevention strategies, such as reduction in salt intake and integration of hypertension care into primary care, may prove beneficial; however, the determinants of hypertension remain to be ascertained. PMID- 12894955 TI - Association of region of residence and immigrant status with hypertension, renal failure, cardiovascular disease, and stroke, among African-American participants in the third National Health and Nutrition Examination Survey (NHANES III). AB - OBJECTIVE: To determine whether current region of residence and immigrant status (born in the United States [US] vs abroad), are associated with the prevalence of hypertension (HTN), uncontrolled HTN, and HTN-related target-organ damage, among African Americans. METHODS: We studied the survey and physical examination data from a nationally representative cohort of 3,369 self-designated Black participants, aged 30-79 years, in the third National Health and Nutrition Examination Survey (NHANES III), which took place during 1988-1994. We calculated the age-adjusted prevalence rates of HTN, uncontrolled HTN, and history of HTN related target-organ damage in US-born northern African Americans, US-born southern African Americans, and foreign-born African Americans. RESULTS: Hypertension (HTN) was more common among southern African-American men and women, compared to northern African-American men and women (42.2% vs 34.1%, P<.002 for men; 42.7% vs 37.2%, P=.02 for women). Uncontrolled HTN was also more common among hypertensive southern African-American women compared to hypertensive northern African-American women (79.8% vs 70.4%, P=.05). Among women, hypertensive Black immigrants had lower rates of HTN-related target-organ damage than either hypertensive US-born southern and northern African Americans (3.3% vs 16.3% and 15.8%, respectively, P=.05). CONCLUSIONS: In this nationally representative cohort, immigrant status and geographic region of residence were associated with HTN prevalence, rates of blood pressure control, and HTN-related target-organ damage. Further examination of environmental exposures, cultural issues, and access to care, factors that can differ between groups, may yield important information about modifiable risk factors associated with HTN and target organ damage. PMID- 12894956 TI - Hypertension, dyslipidemia, body mass index, diabetes and smoking status in Aboriginal Australians in a remote community. AB - OBJECTIVES: Study objectives were: 1) to describe the differences in the prevalence of CHD risk factors between Aboriginal people in a remote community and the general Australian population; and 2) to compare the predicted risks of CHD events between Aboriginal and non-Aboriginal Australians. DESIGN: A cross sectional study. PARTICIPANTS: 681 Aboriginal adults aged 25 to 74 years. RESULTS: Aboriginal young adults had substantially higher prevalence of diabetes compared to non-Aboriginal Australians. The prevalence ratios for diabetes were 12.5, 5.6, 3.2, 1.3, and 0.73 for 25-, 35-, 45-, 55-, and 65- to 74-year-old females, respectively. The corresponding values for males were 12.1, 2.7, 2.9, 0.69, and 0.42. Young females had a higher prevalence of obesity, overweight, and abnormal waist circumference, while males and females 45 years and older tended to have a lower prevalence of overweight and abnormal waist circumference. Compared to the general population, Aboriginal adults had a lower prevalence of abnormal total cholesterol but a higher prevalence of abnormal HDL, triglycerides, hypertension, and smoking. The risk ratios of abnormal total cholesterol for females ages 25-34, 35-44, 45-54, 55-64, and 65-75 years were 0.38, 0.53, 0.48, 0.48, and 0.41, respectively. CONCLUSIONS: Aboriginal people in the remote community experienced different levels of CHD risk predictors from the general Australian population. They had a lower prevalence of abnormal total cholesterol and a higher prevalence of abnormal HDL, smoking, diabetes, and hypertension. PMID- 12894957 TI - Angiotensin converting enzyme inhibitors and beta-blockers in African Americans with heart failure. AB - Previous analyses have implied diminished efficacy of angiotensin converting enzyme inhibitors (ACEI), and equivalent or enhanced efficacy of beta-blockers (BB), in African Americans (AA) with congestive heart failure (CHF), when compared to placebo. These results may have been influenced by lead-time bias, in that AA may not have been entered into the older ACEI trials until late in their CHF course. Our goal was to use a prospective cohort study of 29,686 CHF patients within a single health system to examine the impact on AA mortality of administering ACEI and BB within the first year of CHF diagnosis. Pharmacy claims from 1995-1998 were available for 3353 newly diagnosed CHF patients (39.2% AA; N=1317) within the health maintenance organization. Rates of ACEI and BB use were 46.4% and 54.0%; 43.4% and 28.9%; and 40.7% and 18.6%, for Whites, AA, and other races, respectively. The relative risk reductions (RRR) for ACEI were 68.7%, P<.0001; 52.1%, P<.0001; and -36.3%, P=.56, for Whites, AA, and other races, respectively. The RRR for BB were 59.0%, P<.0001; 34.6%, P=.009; and 74.3%, P=.17, for Whites, AA, and other races, respectively. Age- and gender-adjusted survival rates for AA were significantly enhanced in those taking ACEI, BB, or a combination of the two: P<.001, P=.001, and P=.003, respectively. Although we could not control for selection bias, these data suggest that AA benefit from both ACEI and BB when treatment is initiated within the first year of CHF diagnosis. Future, similar analyses other databases should control for the duration of illness to avoid lead-time bias in AA with CHF. PMID- 12894958 TI - Low-fat diet: effect on anthropometrics, blood pressure, glucose, and insulin in older women. AB - OBJECTIVE: The Women's Health Trial: Feasibility Study in Minority Populations (WHT: FSMP) documented that a low-fat diet was associated with a reduced fat intake in older women of diverse ethnic backgrounds. The purpose of the current study was to examine the effect of the low-fat diet on anthropometric and biochemical variables. DESIGN: Randomized clinical trial in 2,208 postmenopausal women, 50 to 79 years of age. RESULTS: The decrease in fat intake correlated directly with a decrease in body weight (r=.22, P<.001). After 6 months, the intervention group had an average weight loss of 1.8 kg. Body mass index decreased 0.7 kg/m2. Waist circumference decreased 1.8 cm. All of these changes were statistically significant, compared to changes in the control group (P<.01). Changes in systolic (-3.1 mm Hg) and diastolic (-1.1 mm Hg) blood pressures (BP) occurred in the intervention group. The decrease in systolic BP reached statistical significance (P=.02), relative to the control group. Decreases in plasma glucose were small (-0.2 mmol/L) in the intervention group, although there was a trend for difference from the control group (P=.11). Decreases in serum insulin levels were small (-0.5 microIU/mL) in the intervention group, although there was, again, a trend for difference from the control group. CONCLUSIONS: In older White, Black, and Hispanic women, a long-term low-fat dietary intervention was accompanied by modest, but statistically significant, decreases in body weight and anthropometric indices, without any particular attempt being made to reduce calories. Changes in glucose and insulin were small. The long-term biological significance of the glucose and insulin changes is unknown. PMID- 12894959 TI - Culture within the context of care: an integrative review. AB - The purpose of this integrative review was to examine the literature on culturally relevant healthcare interventions, and their effect on health outcomes, in an attempt to determine whether culture matters in the context of healthcare delivery. Research literature on culturally relevant interventions from the past 20 years was reviewed using computerized searches of Medline and CINAHL databases. Results of the review indicate that culturally relevant interventions significantly improve health outcomes for patients with diabetes mellitus (DM), drug addiction, sexually transmitted infections (STIs), and other health problems. It appears that the design of culturally relevant interventions does not require specific knowledge of particular ethnic or cultural groups, but of cross-cultural process principles. Because the studies are highly variable with respect to design and method, it is difficult to isolate which particular aspects of the interventions are specifically associated with favorable outcomes. In addition, few of the studies examined long-term effects of the interventions on outcomes. PMID- 12894960 TI - The effectiveness of a community/academic health center partnership in decreasing the level of blood pressure in an urban African-American population. AB - This study investigated the effectiveness of a community-academic health center partnership, utilizing nurse-supervised indigenous community health workers, in decreasing the blood pressure in an urban African-American population. A four year randomized clinical trial was conducted in the Sandtown-Winchester community, which has an excess prevalence of high blood pressure, in order to test the effectiveness of 2 different levels of intervention intensity on increasing the control of high blood pressure. Community health workers were trained and certified in blood pressure management, monitoring, education and counseling, social support mobilization, and community outreach and follow up. The primary results were a significant decrease in mean systolic and diastolic pressures after both levels of intervention, and a significant increase in the percentage of individuals with controlled high blood pressure. Surprisingly, no differences in results were observed between the 2 levels of intervention intensity. This study supports the use of community-based partnership efforts, and the utilization of indigenous health workers, to enhance the control of high blood pressure in a high-risk, African-American urban population. PMID- 12894961 TI - Social structure, race, and gonorrhea rates in the southeastern United States. AB - OBJECTIVES: We sought to identify characteristics of counties in the southeastern United States associated with endemically high rates of gonorrhea. In particular, we were interested in aspects of race other than the proportion of Blacks in a population, including the potential influence of under-reporting of infections. DESIGN: The associations between the characteristics of counties in 1990, and the rates of reported gonorrhea from 1986 to 1995, were estimated with multivariable logistic regression. SETTING: 14 states in the southeastern region of the United States. PARTICIPANTS: 835 counties and county equivalents in the 14 southeastern states. MAIN OUTCOME MEASURES: The odds of having an endemically high county level rate of gonorrhea. RESULTS: The variables with a strong effect on endemically high rates of gonorrhea included racial residential isolation in the absence of low income dualism (odds ratio [OR]: 210.84, 95% confidence interval [CI]: 19.35, 999.00), and Black-White income dualism in communities with few female-headed households (OR: 4.57, 95% CI: 2.68, 7.80). The percentage of Blacks in the population that was Black had little or no association with the rate of gonorrhea. These estimates were relatively robust when subjected to a sensitivity analysis of potential under-reporting of gonorrhea. CONCLUSIONS: Previous studies have demonstrated that a high percentage of Blacks in a population is the strongest predictor of high rates of gonorrhea. We found, however, that when variables measuring aspects of social structure, such as a race-based income distribution, and de facto residential segregation, were included in the model, the proportion of Blacks no longer had an effect on rates of gonorrhea. Progress in lowering endemically high disease rates will require attention being paid to community racial and class dynamics. PMID- 12894962 TI - Emergency department utilization among Hispanic and African-American under-served patients with type 2 diabetes. AB - OBJECTIVE: To examine emergency department (ED) use among a sample of 349 low income Hispanic and African-American patients with type 2 diabetes. The study used a theoretical model to conceptualize health services utilization as the end product of predisposing, enabling, and need-for-care characteristics. DESIGN: Cross-sectional sampling of consecutive subjects with diabetes attending 7 inner city clinics that provide primary medical care to low-income residents in South Central Los Angeles. PARTICIPANTS: Three hundred and forty-nine patients (Hispanics [67.3%] and African Americans [32.7%]) with type 2 diabetes. RESULTS: Thirty-two percent of this sample reported one or more visits to the ED for medical care within the 12-month period prior to the interviews. Moreover, 11.5% of the patients reported visiting the ED more than once and 4.6% of the patients admitted visiting the ED more than 2 times within the same 12-month period. In a staged path analysis, ED use was significantly predicted by the predisposing variables of younger age, female gender, African-American ethnicity, and less education as well as the enabling variable of ability to pay. In addition, the need variables of insulin injection and number of diabetes-related complications were also significantly related to ED utilization. CONCLUSIONS: Emergency department (ED) utilization is considered to be closely linked to a patient's need-for-care characteristics. However, our study shows that the predisposing characteristics of participants with diabetes explained about the same amount of variance as need-for-care characteristics. The higher-than-anticipated importance of predisposing characteristics suggest that intervention strategies, which specifically target patients with these characteristics, should be a part of any broad approach to reducing ED use among patients with type 2 diabetes. PMID- 12894963 TI - Cancer-related health behaviors and screening practices among Latinos: findings from a community and agricultural labor camp survey. AB - OBJECTIVE: To examine whether cancer-related health behaviors and screening practices differ within a population of Latino adults, including those often missed by cancer surveys. DESIGN: Cross-sectional survey, conducted in 2000. Sample of 461 women and 356 men from the community (75% with unlisted telephones) and 188 men from agricultural labor camps, 18-64 years of age. SETTING: Monterey County, California. OUTCOMES: Six health behaviors and risk factors: obesity, poor nutrition, physical inactivity, high alcohol use, and smoking. Five health practices and screening tests used to detect cervical, breast, and colorectal cancer. RESULTS: Most respondents were born in Mexico, spoke Spanish, and had lived in the United States 10 years or more. In both surveys, more than 60% were overweight including more than 20% who were obese. Men, especially from labor camps, reported high dietary fat intake, low fruit intake, and high alcohol use. For every additional 5 years lived in the United States, the odds of obesity increased 25% for women, and the odds of high-fat/fast food intake and high alcohol use increased 35% and 50%, respectively for labor camp men. Screening rates for cervical and breast cancer were high and met Healthy People 2000 objectives. In contrast, screening rates for colorectal cancer were low; among those 50 and older, approximately 70%-80% of women and men from the community sample and 100% of men from the labor camp sample had never had a blood stool test. Unmarried women, in particular, had poor nutrition and low screening rates. CONCLUSIONS: Cancer control programs for Latinos need a particular focus on weight, nutrition, physical activity, alcohol, and colorectal screening. PMID- 12894964 TI - Establishment of a binational cohort to study Helicobacter pylori infection in children. AB - Chronic Helicobacter (H.) pylori infection, typically of childhood onset, causes upper digestive tract diseases of major impact among socioeconomically marginalized populations. This infection is common in children from ethnic minorities in the United States, and particularly so in immigrant children from Mexico. Prevention measures for H. pylori infection do not yet exist, given limited understanding of what causes either acute or persistent infection. To address this gap, we initiated the Pasitos Cohort Study to follow children from low-income families in the border region that includes El Paso County, Texas, and Ciudad Juarez, Chihuahua. The children were enrolled prior to birth, and are examined at 6-month intervals to observe the natural history of H. pylori infection, and to identify risk factors for acquisition, recurrence, and persistence. This report details the study methods, describes how the cohort was established, and discusses the challenges of compliance with follow up in the border setting. Between April 1998 and October 2000, 1,288 pregnant women were screened for eligibility; 807 of 994 eligible women consented to participate. Birth documentation was obtained for 615 infants, and 472 entered follow up. Successful follow up of this cohort requires resources, including a well-trained, dedicated staff, and incentives, to facilitate and motivate long-term participation. Future findings from this ongoing study will help to fill critical gaps in knowledge regarding the epidemiology of H. pylori infection, and will contribute to the identification of prevention strategies. PMID- 12894965 TI - For the patient. Community health workers help to reduce high blood pressure. PMID- 12894966 TI - For the patient. Scientists study bacteria found more often in children and harmful to stomach. PMID- 12894967 TI - For the patient. Equal health care begins with sensitivity to cultural background of patient. PMID- 12894968 TI - Endovascular interventional magnetic resonance imaging. AB - Minimally invasive interventional radiological procedures, such as balloon angioplasty, stent placement or coiling of aneurysms, play an increasingly important role in the treatment of patients suffering from vascular disease. The non-destructive nature of magnetic resonance imaging (MRI), its ability to combine the acquisition of high quality anatomical images and functional information, such as blood flow velocities, perfusion and diffusion, together with its inherent three dimensionality and tomographic imaging capacities, have been advocated as advantages of using the MRI technique for guidance of endovascular radiological interventions. Within this light, endovascular interventional MRI has emerged as an interesting and promising new branch of interventional radiology. In this review article, the authors will give an overview of the most important issues related to this field. In this context, we will focus on the prerequisites for endovascular interventional MRI to come to maturity. In particular, the various approaches for device tracking that were proposed will be discussed and categorized. Furthermore, dedicated MRI systems, safety and compatibility issues and promising applications that could become clinical practice in the future will be discussed. PMID- 12894969 TI - Use of a quantitative index of beam modulation to characterize dose conformality: illustration by a comparison of full beamlet IMRT, few-segment IMRT (fsIMRT) and conformal unmodulated radiotherapy. AB - A technique is presented for characterizing the degree of modulation in an intensity-modulated beam. It is shown that the modulation increases as dose conformality increases. Full intensity-modulated radiation therapy (IMRT) is compared with a two-weight-per-field technique and with simple geometrically conformal beams. It is suggested that each individual planning problem requires some comparative planning of this type because there is no simple answer to the question of the degree to which IMRT improves dose conformality. This depends on the problem geometry, the dose prescription, the cost function, the number of beams and other planning conditions. A methodology is presented for such comparative planning studies and this is illustrated with the solution of two planning problems. PMID- 12894970 TI - Biological dosimetry of complex ion radiation fields. AB - We report on a set of cell survival experiments performed in complex field combinations of therapeutic 12C ion beams. CHO cells were exposed to the superposition of two or three fields of 12C ions arranged in a similar way to real patient treatments. Two-dimensional survival distributions were measured and compared with the predictions of the TRiP98 treatment planning system. Good agreement was found in general. In particular the method of tissue sparing using dose ramps could be verified. PMID- 12894971 TI - Sequential annealing-gradient Gamma-Knife radiosurgery optimization. AB - Simulated annealing and gradient methods are commonly employed for inverse planning of radiotherapy delivery schemes. Annealing is effective in finding an approximation of the global solution, suffering from slow late convergence and in some cases poor dose homogeneity. Gradient methods converge well but not necessarily to the global minimum. We explored simulated annealing followed by gradient optimization to improve on either method alone, using radiosurgery as the model system. Simulated annealing and gradient inverse planning programs using the same objective function were adapted for radiosurgical optimization. The objective function chosen is a least-squares dose-matching function, with differential weighting of tissues. A simple test target allowing local minima in the objective function was evaluated. Two hundred trials using the gradient method were done. The gradient method approximated the global solution only 12% of the time, commonly finding a local minimum. The annealing-gradient technique converged to the global minimum in 78 out of 80 trials, more efficiently than annealing alone. Dose homogeneity was improved. In conclusion, sequential annealing-gradient optimization can improve on either method alone. The technique may be extensible to radiotherapy inverse planning in general, with benefit expected for problems characterized by slow gradient method convergence and local minima. PMID- 12894972 TI - Ionization chamber dosimetry of small photon fields: a Monte Carlo study on stopping-power ratios for radiosurgery and IMRT beams. AB - Absolute dosimetry with ionization chambers of the narrow photon fields used in stereotactic techniques and IMRT beamlets is constrained by lack of electron equilibrium in the radiation field. It is questionable that stopping-power ratio in dosimetry protocols, obtained for broad photon beams and quasi-electron equilibrium conditions, can be used in the dosimetry of narrow fields while keeping the uncertainty at the same level as for the broad beams used in accelerator calibrations. Monte Carlo simulations have been performed for two 6 MV clinical accelerators (Elekta SL-18 and Siemens Mevatron Primus), equipped with radiosurgery applicators and MLC. Narrow circular and Z-shaped on-axis and off-axis fields, as well as broad IMRT configured beams, have been simulated together with reference 10 x 10 cm2 beams. Phase-space data have been used to generate 3D dose distributions which have been compared satisfactorily with experimental profiles (ion chamber, diodes and film). Photon and electron spectra at various depths in water have been calculated, followed by Spencer-Attix (delta = 10 keV) stopping-power ratio calculations which have been compared to those used in the IAEA TRS-398 code of practice. For water/air and PMMA/air stopping power ratios, agreements within 0.1% have been obtained for the 10 x 10 cm2 fields. For radiosurgery applicators and narrow MLC beams, the calculated s(w,air) values agree with the reference within +/-0.3%, well within the estimated standard uncertainty of the reference stopping-power ratios (0.5%). Ionization chamber dosimetry of narrow beams at the photon qualities used in this work (6 MV) can therefore be based on stopping-power ratios data in dosimetry protocols. For a modulated 6 MV broad beam used in clinical IMRT, s(w,air) agrees within 0.1% with the value for 10 x 10 cm2, confirming that at low energies IMRT absolute dosimetry can also be based on data for open reference fields. At higher energies (24 MV) the difference in s(w,air) was up to 1.1%, indicating that the use of protocol data for narrow beams in such cases is less accurate than at low energies, and detailed calculations of the dosimetry parameters involved should be performed if similar accuracy to that of 6 MV is sought. PMID- 12894973 TI - Convolution/superposition using the Monte Carlo method. AB - The convolution/superposition calculations for radiotherapy dose distributions are traditionally performed by convolving polyenergetic energy deposition kernels with TERMA (total energy released per unit mass) precomputed in each voxel of the irradiated phantom. We propose an alternative method in which the TERMA calculation is replaced by random sampling of photon energy, direction and interaction point. Then, a direction is randomly sampled from the angular distribution of the monoenergetic kernel corresponding to the photon energy. The kernel ray is propagated across the phantom, and energy is deposited in each voxel traversed. An important advantage of the explicit sampling of energy is that spectral changes with depth are automatically accounted for. No spectral or kernel hardening corrections are needed. Furthermore, the continuous sampling of photon direction allows us to model sharp changes in fluence, such as those due to collimator tongue-and-groove. The use of explicit photon direction also facilitates modelling of situations where a given voxel is traversed by photons from many directions. Extra-focal radiation, for instance, can therefore be modelled accurately. Our method also allows efficient calculation of a multi segment/multi-beam IMRT plan by sampling of beam angles and field segments according to their relative weights. For instance, an IMRT plan consisting of seven 14 x 12 cm2 beams with a total of 300 field segments can be computed in 15 min on a single CPU, with 2% statistical fluctuations at the isocentre of the patient's CT phantom divided into 4 x 4 x 4 mm3 voxels. The calculation contains all aperture-specific effects, such as tongue and groove, leaf curvature and head scatter. This contrasts with deterministic methods in which each segment is given equal importance, and the time taken scales with the number of segments. Thus, the Monte Carlo superposition provides a simple, accurate and efficient method for complex radiotherapy dose calculations. PMID- 12894974 TI - Wedge factor dependence with depth and field size for fast neutron beams. AB - The dependence of the wedge factors (WFs) on field size (FS) and depth for a fast neutron beam has been investigated. In a previous study (Popescu et al 1999 Med. Phys. 26 541), a method was presented that allows a simple and accurate way of calculating the wedge-factor dependence on FS and depth in the case of a photon beam. The validity of a similar approach is tested in the present study for neutron beam dosimetry. The clinical neutron therapy system at the University of Washington (UW) has a flattening filter assembly consisting of two filters: a small field filter and a large field filter. Despite this complication, the approach presented in Popescu et al (1999 Med. Phys. 26 541) can be used to describe the WF dependence on FS and depth (d). PMID- 12894975 TI - High dose-rate brachytherapy source localization: positional resolution using a diamond detector. AB - A potential real-time source position verification process for high dose-rate (HDR) brachytherapy treatment is described. This process is intended to provide immediate confirmation that a treatment is proceeding according to plan, so that corrective action can be taken if necessary. We show that three dosimeters are in principle sufficient and demonstrate the feasibility of the process using a diamond detector and an Ir-192 source. An error analysis including all identified sources of error shows that this detector is capable of locating the distance to the source to within 2 mm for distances up to 12 cm. This positional accuracy is less than the diameter of typical HDR catheters indicating that a diamond detector can be used to accurately determine the distance to the source. The uncertainty in the distance is found to increase with distance. PMID- 12894976 TI - Theoretical cross sections for electron collisions in water: structure of electron tracks. AB - To understand what happens when biological matter is irradiated needs a detailed knowledge of the microscopic distribution of interactions and especially of the energy deposited in irradiated matter. Monte Carlo event-by-event simulations are particularly suitable for this task. However, the development of these track structure codes necessitates accurate interaction cross sections for all the electronic processes: ionization, excitation and elastic scattering. In these conditions, we have recently developed a Monte Carlo code for electrons in water, this latter being commonly used to simulate the biological medium. All the electronic processes are studied in detail via theoretical differential and total cross-section calculations. The purpose of this work is to make an inter comparison of our cross sections with those used in the electron track-structure codes developed in the literature, and to compare macroscopic quantities such as stopping powers and mean energy transfer distributions to available experimental data and/or to theoretical predictions in liquid water. PMID- 12894977 TI - Effect of spurt duration on the heat transfer dynamics during cryogen spray cooling. AB - Although cryogen spray cooling (CSC) is used to minimize the risk of epidermal damage during laser dermatologic surgery, optimization of the current cooling approach is needed to permit the safe use of higher light doses, which should improve the therapeutic outcome in many patients. The objective of this study was to measure the effect of spurt duration (delta t) on the heat transfer dynamics during CSC using a model skin phantom. A fast-response temperature sensor was constructed to record the changes in surface temperature during CSC. Temperature measurements as a function of delta t at two nozzle-to-skin distances (z = 50 and 20 mm) were performed. The average surface heat fluxes (q) and heat transfer coefficients (h) for each delta t were computed using an inverse heat conduction problem algorithm. It was observed that q undergoes a marked dynamic variation during the entire delta t, with a maximum heat flux (qc) occurring early in the spurt (5-15 ms), followed by a quick decrease. The estimated qc vary from 450 to 600 kW m(-2), corresponding to h maxima of 10 and 17-22 kW m(-2) K(-1) for z = 50 and 20 mm, respectively. For z = 50 mm, spurts longer than 40 ms do not increase the total heat removal (Q) within the first 200 ms. However, for z = 20 mm, delta t longer than 100 ms are required to achieve the same Q. It is shown that the heat transfer dynamics and the time it takes to reach qc during CSC can be understood through classic boiling theory as a transition from transient to nucleate boiling. Based on the results of this model skin phantom, it is shown that spurts longer than 40 ms have a negligible impact on both q and Q within clinically relevant cooling times (10-100 ms). PMID- 12894978 TI - Measuring the elastic modulus of ex vivo small tissue samples. AB - Over the past decade, several methods have been proposed to image tissue elasticity based on imaging methods collectively called elastography. While progress in developing these systems has been rapid, the basic understanding of tissue properties to interpret elastography images is generally lacking. To address this limitation, we developed a system to measure the Young's modulus of small soft tissue specimens. This system was designed to accommodate biological soft tissue constraints such as sample size, geometry imperfection and heterogeneity. The measurement technique consists of indenting an unconfined small block of tissue while measuring the resulting force. We show that the measured force-displacement slope of such a geometry can be transformed to the tissue Young's modulus via a conversion factor related to the sample's geometry and boundary conditions using finite element analysis. We also demonstrate another measurement technique for tissue elasticity based on quasi-static magnetic resonance elastography in which a tissue specimen encased in a gelatine agarose block undergoes cyclical compression with resulting displacements measured using a phase contrast MRI technique. The tissue Young's modulus is then reconstructed from the measured displacements using an inversion technique. Finally, preliminary elasticity measurement results of various breast tissues are presented and discussed. PMID- 12894979 TI - Neuronavigation accuracy dependence on CT and MR imaging parameters: a phantom based study. AB - Clinical benefits from neuronavigation are well established. However, the complexity of its technical environment requires a careful evaluation of different types of errors. In this work, a detailed phantom study which investigates the accuracy in a neuronavigation procedure is presented. The dependence on many different imaging parameters, such as field of view, slice thickness and different kind of sequences (sequential and spiral for CT, T1 weighted and T2-weighted for MRI), is quantified. Moreover, data based on CT images are compared to those based on MR images, taking into account MRI distortion. Finally, the contributions to global accuracy coming from image acquisition, registration and navigation itself are discussed. Results demonstrate the importance of imaging accuracy. Procedures based on CT proved to be more accurate than procedures based on MRI. In the former, values from 2 to 2.5 mm are obtained for 95% fractiles of cumulative distribution of Euclidean distances between the intended target and the reached one while, in the latter, the measured values range from 3 to 4 mm. The absence of imaging distortion proved to be crucial for registration accuracy in MR-based procedures. PMID- 12894980 TI - Why should breast tumour detection go three dimensional? AB - Although x-ray mammography is widely developed for breast tumour detection, it suffers from spatial superposition in its two-dimensional (2D) representation of a three-dimensional (3D) breast structure. Accordingly, 3D breast imaging, such as cone-beam computed tomography (CT), arises at the historic moment. In this paper, we theoretically elucidate the spatial superposition effect associated with x-ray mammography on breast tumour detection. This explanation is based on the line integral of x-ray traversing a composite breast model. As a result, we can characterize the difficulty of detecting small tumours in terms of local intensity contrast in x-ray images. In comparison, we also introduce cone-beam CT breast imaging for 3D breast volume representation, which offers advantages for breast mass segmentation and measurement. The discussion is demonstrated with an experiment with a breast surgical specimen. In conclusion, we strongly believe that 3D volumetric representation allows for more accurate breast tumour detection. PMID- 12894981 TI - Classification of breast masses in ultrasonic B scans using Nakagami and K distributions. AB - Classification of breast masses in greyscale ultrasound images is undertaken using a multiparameter approach. Five parameters reflecting the non-Rayleigh nature of the backscattered echo were used. These parameters, based mostly on the Nakagami and K distributions, were extracted from the envelope of the echoes at the site, boundary, spiculated region and shadow of the mass. They were combined to create a linear discriminant. The performance of this discriminant for the classification of breast masses was studied using a data set consisting of 70 benign and 29 malignant cases. The Az value for the discriminant was 0.96 +/- 0.02, showing great promise in the classification of masses into benign and malignant ones. The discriminant was combined with the level of suspicion values of the radiologist leading to an Az value of 0.97 +/- 0.014. The parameters used here can be calculated with minimal clinical intervention, so the method proposed here may therefore be easily implemented in an automated fashion. These results also support the recent reports suggesting that ultrasound may help as an adjunct to mammography in breast cancer diagnostics to enhance the classification of breast masses. PMID- 12894982 TI - Accuracy assessment protocols for electromagnetic tracking systems. AB - Electromagnetic tracking systems have found increasing use in medical applications during the last few years. As with most non-trivial spatial measurement systems, the complex determination of positions and orientations from their underlying raw sensor measurements results in complicated, non-uniform error distributions over the specified measurement volume. This makes it difficult to unambiguously determine accuracy and performance assessments that allow users to judge the suitability of these systems for their particular needs. Various assessment protocols generally emphasize different measurement aspects that typically arise in clinical use. This can easily lead to inconclusive or even contradictory conclusions. We examine some of the major issues involved and discuss three useful calibration protocols. The measurement accuracy of a system can be described in terms of its 'trueness' and its 'precision'. Often, the two are strongly coupled and cannot be easily determined independently. We present a method that allows the two to be disentangled, so that the resultant trueness properly represents the systematic, non-reducible part of the measurement error, and the resultant precision (or repeatability) represents only the statistical, reducible part. Although the discussion is given largely within the context of electromagnetic tracking systems, many of the results are applicable to measurement systems in general. PMID- 12894983 TI - Dielectrophoretic assay of bacterial resistance to antibiotics. AB - The dielectrophoretic collection spectra of antibiotic-sensitive and antibiotic resistant strains of Staphylococcus epidermidis have been determined. These indicate that in the absence of antibiotic treatment there is a strong similarity between the dielectric properties of sensitive and resistant strains, and that there is a significant difference between the sensitive strains before and after treatment with the antibiotic streptomycin after 24 h exposure. This method offers possibilities for the assessment of bacterial resistance to antibiotics. PMID- 12894984 TI - Comprehensive quality assurance for stereotactic radiosurgery treatments. AB - We have used a commercially available high precision Lucy phantom to perform comprehensive quality assurance for stereotactic radiosurgery treatments. The quantitative evaluation of system uncertainties included imaging, planning and treatment delivery systems. The quality assurance tests showed that the well defined targets were identified to within +/-1 mm in all the imaging modalities. The pre-known target volumes were reproduced within 2 cm3 in both MR and CT. The planned target was delivered within 2% of the prescribed dose and to within 2 mm accuracy. The inaccuracy in the isocentre position at the Linac was less than 1.2 mm. The maximum error observed in the depth helmet was 0.5 mm and the overall uncertainty was within 0.23 mm. We have also established a quality assurance program based on the study and proposed the tolerance and the frequency of the tests required to be carried out. The tests were carried out using a Radionics planning system and delivered on a Varian Clinac 2100 linear accelerator machine. These tests also established a base line for future comparisons. PMID- 12894985 TI - Changes in the diffraction pattern of hair resulting from mechanical damage can occlude the changes that relate to breast cancer. PMID- 12894986 TI - Factors that mediate and modulate androgen action. AB - Androgens mediate a wide range of processes during embryogenesis and in the adult. In mammals, although a number of steroids can be shown to exert androgenic effects using in vitro and in vivo assays, testosterone and its 5alpha reduced metabolite, 5alpha-dihydrotestosterone (DHT) are considered to represent the principal physiologic androgens. Furthermore, although the effects that androgens exert differ widely among different tissues and cell types, genetic and biochemical data suggest that these effects are mediated via the protein products of a single androgen receptor gene, which is encoded on the X-chromosome in mammals. PMID- 12894987 TI - The pathogenesis of alopecia areata in rodent models. AB - Rodent models of human disease provide an important tool in the investigation of genetic and environmental activation factors, disease pathogenesis, and the development of new and improved treatments. Up to 20% of aged C3H/HeJ mice and 70% of Dundee Experimental Bald Rats (DEBR) develop alopecia areata (AA), a nonscarring, inflammatory hair loss disease with a suspected autoimmune pathogenesis. These rodent models are currently employed in determining the genetic basis of AA, understanding the mechanisms of disease initiation and progression, and defining potential endogenous and environmental influences. Induction of AA by skin graft transfer between affected and unaffected mice has been employed to examine skin and immune system changes during AA pathogenesis. Manipulation of inflammatory cells in vivo indicates AA is primarily a cell mediated disease with auto-antibody production as a secondary event. Whether the AA activating factors are exogenous or endogenous antigens, or involve normal or aberrant epitope expression remains to be elucidated. However, current research suggests a self contained disease cycle involving four key events: (1) Failure of the putative anagen stage hair follicle immune privilege and exposure of hair follicle located AA inciting epitopes to the immune system; (2) Antigen presentation, costimulation, and activation of responsive lymphocytes by antigen presenting cells; (3) Activated inflammatory cell migration to, and infiltration of, hair follicles; (4) The subsequent disruptive actions of the inflammatory cell infiltrate on the hair follicles. Each of these events is vulnerable to therapeutic intervention, and rodent models will be fundamentally involved in developing new treatments for AA. PMID- 12894988 TI - Alopecia areata: treatment of today and tomorrow. AB - It is the aim of this article to review and appraise available data on treatments for alopecia areata (AA) according to the demands of evidence based medicine. Studies evaluating the efficacy of a treatment for AA should include appropriate controls, use cosmetically acceptable hair regrowth as a parameter for treatment success, include patients with AA totalis, universalis or extensive patchy AA, and exclude patients suffering from AA for less than 3 months. Moreover, the treatment must be safe over a prolonged period of time. Among the various therapeutic approaches presently available for AA, only treatment with contact sensitizers such as diphenylcyclopropenone or squaric acid dibutylester has been shown to be effective in studies that fulfill these criteria. Improved future treatments may be immunosuppressive or immunomodulatory targeting of the autoimmune pathogenesis of AA, or they may otherwise protect hair follicles from the injurious effects of inflammation. Such possible future therapeutic approaches include the incorporation of immunomodulatory agents into liposomes as an improved vehicle; inhibition of apoptosis mediated by the Fas-FasL system; inhibition of the lymphocyte homing receptor CD44v10; induction of tolerance. PMID- 12894989 TI - Update on cicatricial alopecia. AB - Cicatricial alopecia is an enigmatic group of hair disorders linked by the potential permanent loss of scalp hair follicles in involved areas. Progress in our understanding and treatment of these disorders has been stymied by the lack of clear diagnostic criteria for the current terms used to describe the various hair loss entities. Since all of these conditions evolve as the hair is destroyed or replaced, diagnosis is further made difficult by a lack of clinical and pathologic "snapshots" over the evolution of each disorder. Without some acceptance of general clinical and histological presentations in the early, mid and late stage of these disorders, one cannot begin to explore ways to make the diagnosis at a very early stage before significant follicular destraction has occurred (making the clinical diagnosis obvious) and when the damage is potentially repairable or progression preventable. PMID- 12894990 TI - Use of finasteride in the treatment of men with androgenetic alopecia (male pattern hair loss). AB - Finasteride, a type 2-selective 5alpha-reductase inhibitor, was approved in 1997 as the first oral pharmacologic therapy for the treatment of men with androgenetic alopecia (AGA; male pattern hair loss). Originally developed for the treatment of men with benign prostatic hyperplasia (BPH) at a dose of 5 mg/day, finasteride has a well-established, excellent safety profile. Subsequent studies demonstrated that finasteride was an effective treatment for men with AGA at an optimal dose of 1 mg/day. This report summarizes the published peer-reviewed literature on the use of finasteride in the treatment of men with AGA, including the data on long-term (5 years) use of finasteride in a placebo-controlled clinical trial environment. PMID- 12894991 TI - Androgenetic alopecia in women. AB - Androgenetic alopecia (AGA), also known in women as female pattern hair loss, is caused by androgens in genetically susceptible women and men. The thinning begins between ages 12 and 40 years, the inheritance pattern is polygenic, and the incidence is the same as in men. In susceptible hair follicles, dihydrotestosterone binds to the androgen receptor, and the hormone-receptor complex activates the genes responsible for the gradual transformation of large terminal follicles to miniaturized follicles. Both young women and young men with AGA have higher levels of 5alpha reductase and androgen receptor in frontal hair follicles compared to occipital follicles. At the same time, young women have much higher levels of cytochrome p-450 aromatase in frontal follicles than men who have minimal aromatase, and women have even higher aromatase levels in occipital follicles. The diagnosis of AGA in women is supported by early age of onset, the pattern of increased thinning over the frontal/parietal scalp with greater density over the occipital scalp, retention of the frontal hairline, and the presence of miniaturized hairs. Most women with AGA have normal menses and pregnancies. Extensive hormonal testing is usually not needed unless symptoms and signs of androgen excess are present such as hirsutism, severe unresponsive cystic acne, virilization, or galactorrhea. Topical minoxidil solution is the only drug available for promoting hair growth in women with AGA. Efficacy has been shown in double-blind studies using hair counts and hair weight. PMID- 12894992 TI - Hair follicle stem cells. AB - The workshop on Hair Follicle Stem Cells brought together investigators who have used a variety of approaches to try to understand the biology of follicular epithelial stem cells, and the role that these cells play in regulating the hair cycle. One of the main concepts to emerge from this workshop is that follicular epithelial stem cells are multipotent, capable of giving rise not only to all the cell types of the hair, but also to the epidermis and the sebaceous gland. Furthermore, such multipotent stem cells may represent the ultimate epidermal stem cell. Another example of epithelial stem cell and transit amplifying cell plasticity, was the demonstration that adult corneal epithelium, under the influence of embryonic skin dermis could form an epidermis as well as hair follicles. With regards to the location of follicular epithelial stem cells, immunohistochemical and ultrastructural data was presented, indicating that cells with stem cell attributes were localized to the prominent bulge region of developing human fetal hair follicles. Finally, a new notion was put forth concerning the roles that the bulge-located stem cells and the hair germ cells played with respect to the hair cycle. PMID- 12894993 TI - Animal and in vitro models for the study of hair follicles. AB - Since the way in which the hair follicle functions is not well understood, many hair disorders are poorly controlled. A range of in vitro and in vivo models have therefore been developed to investigate the cell biological and biochemical mechanisms involved in the organization of this complex tissue. These range from cultures of a single cell type, such as those of the regulatory, mesenchyme derived dermal papilla, through organ culture of isolated follicles to natural or genetically manipulated animal models. Each system has advantages and disadvantages for studying particular aspects of follicular function and some are potentially useful for the development of novel treatments for hair disorders. PMID- 12894994 TI - Molecular control of epithelial-mesenchymal interactions during hair follicle cycling. AB - Epithelial-mesenchymal interactions play pivotal roles in the morphogenesis of many organs and various types of appendages. During hair follicle development, extensive interactions between two embryologically different hair follicle compartments (epidermal keratinocytes and dermal papilla fibroblasts) lead to the formation of the hair shaft-producing mini-organ that shows cyclic activity during postnatal life with periods of active growth, involution and resting. During the hair cycle, the epithelium and the mesenchyme are regulated by a distinct set of molecular signals that are unique for every distinct phase of the hair cycle. In telogen hair follicles, epithelial-mesenchymal interactions are characterized by a predominance of inhibitory signals that retain the hair follicle in a quiescent state. During anagen, a large variety of growth stimulatory pathways are activated in the epithelium and in the mesenchyme, the coordination of which are essential for proper hair fiber formation. During catagen, the termination of anagen-specific signaling interactions between the epithelium and the mesenchyme leads to apoptosis in the hair follicle epithelium, while activation of selected signaling pathways promotes the transition of the dermal papilla into a quiescent state. The signaling exchange between the follicular epithelium and the mesenchyme is modulated by proteoglycans, such as versican, which may significantly enhance or reduce the biological activities of secreted growth stimulators. However, additional research will be required to bridge the gap between our current understanding of mechanisms underlying epithelial-mesenchymal interactions in hair follicles and the potential clinical application of growth modulators involved in those interactions. Further progress in this area of research will hopefully lead to the development of new drugs for the treatment of hair growth disorders. PMID- 12894995 TI - Morphological approach to hair disorders. AB - The Workshop on the morphological approach to hair disorders brought together a group of clinicians involved in hair biology research. Six speakers spoke on a range of topics that can be grouped broadly into a central theme. It summarizes the evolution of medical research. The section by Tosti and coworkers describes a patient with a new unique syndrome. The section by Ferrando and colleagues provides a framework in which patients with rare hair disorders can be classified. The section by Whiting tries to define the normal anatomy of the hair follicle and both horizontal and vertical sections. It is only when normal anatomy has been absolutely defined that pathological deviations can be recognized. The section by Sinclair and coworkers attempts to estimate the reliability of histological diagnosis so that its true value of pathology can be recognized. The section by Zlotogorski and coworkers shows how accurate clinical and histological definition of disease acts as the cornerstone for gene discovery techniques. Once a causative mutation is found and a gene product identified, then the biological consequences of the altered protein product can be studied and the impact of the abnormal molecular function on hair biology can be understood. It is hoped that improved understanding of hair disease will then lead to useful therapeutic interventions. The final section by Leroy and Van Neste highlights the difficulties of evaluating therapeutic interventions in hair loss disease and proposes a new technique. PMID- 12894996 TI - A potential suppressor of TGF-beta delays catagen progression in hair follicles. AB - TGF-beta plays important roles in the induction of catagen during the hair cycle. We examined whether TGF-beta2 could activate a caspase in human hair follicles. Using active caspase-9 and -3 specific antibodies, we found that TGF-beta2 activated these caspases in two regions, the lower part of the hair bulb and the outer layer of the outer root sheath. In addition, we searched for a plant extract that can effectively suppress TGF-beta action. We found that an extract of Hydrangea macrophylla reduced synthesis of a TGDbeta-inducible protein. We confirmed that the extract has a potential to promote hair elongation in the organ culture system. Furthermore, it delayed in vivo progression of catagen in a mouse model. Our results suggest that the induction of catagen by TGF-beta is mediated via activation of caspases and that a suppressor of TGF-beta could be effective in preventing male pattern baldness. PMID- 12894997 TI - Identification of androgen-inducible TGF-beta1 derived from dermal papilla cells as a key mediator in androgenetic alopecia. AB - We attempted to establish a coculture model of human dermal papilla cells (DPCs) from androgenetic alopecia (AGA) and keratinocytes (KCs) to study the pathomechanism of AGA. Since expression of mRNA for the androgen receptor (AR) decreased during subcultivation of DPCs in vitro, we transiently transfected the AR expression vector into the DPCs and cocultured them with KCs. In this coculture, androgen inhibited the growth of KCs by 50%, indicating that the DPCs produce diffusible growth suppressive factors into the medium in an androgen dependent manner. Since recently increasing evidence has shown the importance of transforming growth factor-beta1 (TGF-beta1) in hair growth, we further examined the concentration of TGF-beta1 in this coculture medium after androgen treatment by ELISA assays. The results showed that androgen treatment increased the secretion of TGF-beta1 into the conditioned medium. Moreover, neutralizing anti TGF-beta1 antibody reversed the inhibition of KC proliferation. Thus, we suggest that androgen-inducible TGF-beta1 derived from DPCs mediates hair growth suppression in AGA. PMID- 12894998 TI - Molecular mechanisms of chemotherapy-induced hair loss. AB - Hair loss (alopecia) is a much-feared side-effect of many chemotherapy protocols and is one of the most psychological devastating aspects of cancer therapy. So far, no satisfactory strategy for suppressing chemotherapy-induced alopecia is at hand. During the last decade, some progress in understanding molecular mechanisms of chemotherapy-induced hair loss has been achieved using rodent models. However, the pathobiology of the response of human hair follicle to chemotherapy remains largely unknown. Here, we review molecular mechanisms that control apoptosis in the hair follicle induced by chemotherapy and delineate the basic strategy for pharmacological inhibition of this devastating side-effect of cancer treatment. We focus on the roles of p53 and its target genes that are essential in mediating responses of hair follicle cells. We assume that local pharmacological inhibition of p53 activity may serve as an effective treatment to prevent chemotherapy induced hair loss. Sufficient pharmacological inhibition of chemotherapy-induced hair loss may require a combination of inhibitors to block complementary or redundant pathways of apoptosis in hair follicles. PMID- 12894999 TI - Fate of melanocytes during development of the hair follicle pigmentary unit. AB - During hair follicle morphogenesis, melanocyte precursors migrate into developing hair follicles and give rise to differentiated melanocytes that actively produce and transport pigment into the keratinocytes that form the hair shaft; however, patterns of melanocyte proliferation and differentiation during formation of the hair pigmentation unit remain to be elucidated. Using multicolor confocal microscopy and double immunofluorescence of melanogenic proteins (tyrosinase related proteins 1 and 2, tyrosinase) and the proliferative marker Ki67, we have studied melanocyte development in C57BL/6 mouse embryonic hair follicles. Proliferating melanocyte precursors (tyrosinase-related protein-2/Ki67+ cells) are seen in the hair follicles at stages 1-2 of morphogenesis, as follicular invagination begins. In stage 3-4 hair follicles, the majority of intrafollicular melanocytes remain tyrosinase-related protein-2+ and Ki67+, whereas some located adjacent to the forming dermal papilla begin to express tyrosinase-related protein-1, an early marker of differentiation. Melanin granules appear in stage 5 hair follicles coincident with tyrosinase expression in nonproliferating tyrosinase-related protein-2+/tyrosinase-related protein-1+ melanocytes. Stage 6 8 hair follicles, those actively producing hair, show nonproliferating tyrosinase related protein-2+ melanocytes in the bulge area, tyrosinase-related protein 2+/tyrosinase-related protein-1+ melanocytes in the outer root sheath, and tyrosinase-related protein-2+/tyrosinase-related protein-1+/tyrosinase+ melanocytes above the dermal papilla. These data suggest that melanocyte precursor cells proliferate extensively at the onset of follicle development. Progeny of these cells migrate down the developing follicle, differentiating further until reaching the area immediately above the dermal papilla, where fully differentiated nonproliferative melanin-producing melanocytes persist, contributing pigment to the growing hair shaft. PMID- 12895000 TI - Plasticity and cytokinetic dynamics of the hair follicle mesenchyme during the hair growth cycle: implications for growth control and hair follicle transformations. AB - Hair fiber production is the macroscopic end-point of a highly complex set of interactions between the hair follicle's epithelial and mesenchymal components. The nature of this relationship is largely set during hair follicle morphogenesis, but is dramatically revisited in the adult during the unique tissue remodeling events required for hair follicle cycling. Whereas significant attention has focused on the fate of the hair follicle epithelium during these events, associated changes in hair follicle fibroblast subpopulations remain unclear. Here, we present a speculative review that represents a critical and innovative synthesis of the current literature and summarizes a recently submitted original study by the authors, on the nature of hair cycle-dependent fibroblast dynamics and on how perturbations thereof may lead to several clinical manifestations of altered human hair growth. PMID- 12895001 TI - Cytokines and signal transduction pathways mediated by anthralin in alopecia areata-affected Dundee experimental balding rats. AB - Although many therapeutic modalities have been tested on alopecia areata, patient outcomes have been disappointing. Use of animal models would help to develop more efficient therapies as well as understanding therapeutic mechanisms. We have demonstrated that 0.1% topical anthralin ointment is 100% effective in restoring follicular activity in Dundee experimental balding rats. This is the most promising topical treatment for Dundee experimental balding rats among the therapeutic agents tested on this model. Various cytokines have been shown to be associated with the pathogenesis of alopecia areata. To test whether any of these cytokines might be modulated by anthralin, an RNase protection assay and the real time polymerase chain reaction were performed to compare their expression between anthralin-treated and control skins. These experiments showed that expression of tumor necrosis factor-alpha and interferon-gamma was inhibited by anthralin, whereas expression of interleukin-1alpha/beta and their receptor antagonist, interleukin-1Ra, and interleukin-10 was stimulated by anthralin. In addition, using an antibody-based multi-immunoblotting technique, we found that certain signaling regulatory proteins were modulated by anthralin. Their potential roles in reversing the autoimmune-arrested follicular activity in Dundee experimental balding rats are discussed. PMID- 12895002 TI - Label-retaining cells (presumptive stem cells) of mice vibrissae do not express gap junction protein connexin 43. AB - We investigated whether connexin 43, a gap junction protein present in human epidermis and mouse hair follicle, can serve as a negative marker for keratinocyte stem cells. Experiments carried out in mouse pelage and vibrissae hair follicles demonstrated that most of the slowly cycling cells, detected as label-retaining cells, do not express connexin 43. In humans, cells with immunohistochemically undetectable levels of connexin 43 are found in the epidermal basal layer of neonatal foreskin, and in the follicular bulge region. About 10% of the basal keratinocytes are connexin 43 negative, as determined by flow cytometry. These cells are uniformly small and low in granularity suggesting that presumptive keratinocyte stem cells can be identified and separated based on connexin 43 expression. PMID- 12895003 TI - Characterization of human keratin-associated protein 1 family members. AB - Keratin-associated proteins are involved in the formation of the cross-linked network of the keratin-intermediate filament proteins that support hair fibers. In recent years, several keratin-associated protein genes have been identified and become an attractive topic in hair research. More recently, we isolated two cDNA encoding novel members of the human keratin-associated protein 1 family (human keratin-associated protein 1.6 and human keratin-associated protein 1.7), and described their expression in the hair follicle by RNA in situ hybridization. A comparison of human keratin-associated protein 1.6 and human keratin-associated protein 1.7 with other human keratin-associated protein 1 members revealed that keratin-associated protein 1 proteins are fundamentally composed of five distinct domains, and that they can be classified primarily by a striking variation in double cysteine-containing pentapeptide repeats in the repetitive I domain. The sum of the data analyzed suggests that human keratin-associated protein 1 family genes may have arisen mainly through gene duplication of the cysteine-repeat motifs during evolution. PMID- 12895004 TI - The distribution of estrogen receptor beta is distinct to that of estrogen receptor alpha and the androgen receptor in human skin and the pilosebaceous unit. AB - Both estrogens and androgens play important parts in skin and hair physiology, although studies of estrogen action in human skin have been rather limited. Recently, a second estrogen receptor (beta) has been identified in many nonclassical target tissues, including androgen-dependent tissues. Therefore, we have revisited the role of estrogens in human skin and hair by comparing the pattern of expression by immunohistochemistry for both estrogen receptors (alpha and beta) and the androgen receptor. Immunolocalization of androgen receptors was only seen in hair follicle dermal papilla cells and the basal cells of the sebaceous gland. Little specific staining of estrogen receptor alpha was seen anywhere except the sebaceous gland. In contrast estrogen receptor beta was highly expressed in epidermis, blood vessels, and dermal fibroblasts, whereas in the hair follicle it was localized to nuclei of the outer root sheath, epithelial matrix, and dermal papilla cells. Serial sections also showed strong nuclear expression of estrogen receptor beta in the cells of the bulge, whereas neither estrogen receptor alpha or androgen receptor was expressed. In the sebaceous gland, estrogen receptor beta was expressed in both basal and partially differentiated sebocytes in a similar pattern to estrogen receptor alpha. There was no obvious difference in the expression of either estrogen receptor in male or female nonbalding scalp skin. The results of this immunohistochemical study propose that estrogen receptor beta and not estrogen receptor alpha is the main mediator of estrogen action in human skin and the hair follicle. Further studies with androgen-dependent skin are required to determine whether estrogen receptor beta has a regulatory role on androgen receptor expression in the hair follicle in parallel with its role in other androgen-dependent tissues. PMID- 12895005 TI - Fas-deficient C3.MRL-Tnfrsf6(lpr) mice and Fas ligand-deficient C3H/HeJ Tnfsf6(gld) mice are relatively resistant to the induction of alopecia areata by grafting of alopecia areata-affected skin from C3H/HeJ mice. AB - Alopecia areata is suspected to be a T cell-mediated autoimmune disease of the hair follicle, where Fas is expressed on hair follicles and Fas ligand on perifollicular infiltrates. To elucidate whether the Fas/Fas ligand pathway is of pathogenetic significance in alopecia areata, we investigated whether alopecia areata can be induced in Fas-deficient and Fas ligand-deficient mice and whether alopecia areata develops in Fas-deficient and Fas ligand-deficient skin. Therefore, we induced alopecia areata by grafting alopecia areata-affected C3H/HeJ mouse skin on to C3H/HeJ mice (control), on to Fas ligand-deficient C3H/HeJ-Tnfsf6(gld) mice or Fas-deficient C3.MRL-Tnfrsf6(lpr) mice. All control mice developed alopecia areata, whereas no Fas-deficient mice showed hair loss and two of seven Fas ligand-deficient mice developed only transitory, limited alopecia areata. Moreover, skin from C3H/HeJ mice (control), C3H/HeJ-Tnfsf6(gld) mice, and C3.MRL-Tnfrsf6(lpr) mice was grafted on to C3H/HeJ mice with extensive alopecia areata. Skin grafts from control mice developed hair loss, whereas Fas deficient and Fas ligand-deficient skin grafts were spared from alopecia areata. Terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling and immunofluorescence studies revealed an increased number of apoptotic cells and expression of Fas on hair follicles as well as expression of Fas ligand on cells of the perifollicular infiltrate in C3H/HeJ mice with alopecia areata, whereas in Fas-deficient and Fas ligand-deficient mice apoptotic cells were virtually absent in hair follicles. The results suggest that the Fas/Fas ligand pathway plays an important pathogenetic role in alopecia areata. PMID- 12895006 TI - TrichoScan: a novel tool for the analysis of hair growth in vivo. AB - Hair loss or hair thinning is a common complaint in clinical dermatology, and patients seeking advice for hair loss are not necessarily bald. Because the effects of treatment attempts are hard to measure, there is need for a sensitive tool to monitor hair loss and treatment responses. Such a method must be able to analyze the biologic parameters of hair growth, which are: (i) hair density (n per cm2); (ii) hair diameter (microm); (iii) hair growth rate (mm per day); and (iv) anagen/telogen ratio. Herein we present the TrichoScan as a method that combines epiluminescence microscopy with automatic digital image analysis for the measurement of human, and potentially animal hair, in situ. The TrichoScan is able to analyze all four parameters of hair growth with a so-called intraclass correlation of approximately 91% within the same TrichoScan operator and an intraclass correlation of approximately 97% for different TrichoScan operators. The application of the technique is demonstrated by comparing the hair parameters in individuals without apparent hair loss, men with untreated androgenetic alopecia, and men after treatment with finasteride (1 mg per day). We were able to detect a significant increase in hair counts and cumulative hair thickness 3 and 6 mo after treatment. Advantages of the TrichoScan are that it can be used for clinical studies to compare placebo versus treatment, to compare different capacities of hair growth promoting substances, to study androgenetic alopecia and other forms of diffuse hair loss, and to study the effects of drugs and laser treatment on hypertrichosis and hirsutism. PMID- 12895007 TI - Phototrichogram analysis of Japanese female subjects with chronic diffuse hair loss. AB - Hair growth patterns of 101 Japanese female subjects with diffuse, chronic hair loss and 58 healthy Japanese female volunteers were categorized into subgroups using noninvasive quantitative methods after determining the key parameters of hair growth. Phototrichogram was performed at 0 and 48 h after clipping hairs in the parietal region of the scalp. Shaft diameters of the excised hairs were then measured. Multiple regression analysis indicated that hair densities, hair diameters, short hair ratios, and hair growth rates, but not anagen hair ratios, were significant, in order of decreasing importance, for grading female diffuse alopecia. Using cluster analysis, hair growth patterns among subjects complaining of diffuse hair loss were divided into six abnormal groups (n=60), two borderline groups (n=21), and one normal group (n=20). The control subjects judged to be normal by macroscopic observation, actually included two subjects with borderline hair growth patterns and one abnormal subject. Most of the abnormal groups shared features of female androgenetic alopecia. Hair patterns showing a decrease in hair density but without vellus hair change, however, emerged as the most prevalent and distinct pattern of chronic diffuse hair loss among the Japanese female subjects. The phototrichogram, combined with the measurement of hair diameters, is an accurate tool for assessing hair growth patterns, especially in detecting the slight changes indicative of the early phase of diffuse alopecia. PMID- 12895008 TI - Progressive patterned scalp hypotrichosis, with wiry hair, onycholysis, and intermittently associated cleft lip and palate: clinical and genetic distinction from Marie Unna. AB - Marie Unna hereditary hypotrichosis has been described in over a dozen families since 1924. Features include scant or no eyebrows at birth, the development of firm wiry hair in the first few years of life followed by a progressive patterned scalp alopecia in the second or third decade. This is associated with generalized hypotrichosis of the body and the condition is nonsyndromic. We have identified a novel form of autosomal dominant ectodermal dysplasia that resembles Marie Unna hereditary hypotrichosis in a family of 23 members over four generations. Affected individuals have patterned hair loss and associated hair shaft dystrophy similar to that seen in Marie Unna hereditary hypotrichosis. It differs from Marie Unna hereditary hypotrichosis by an absence of signs of affectation at birth, relative sparing of body hair, distal onycholysis, and intermittent cosegregation with autosomal dominant cleft lip and palate. Linkage studies to the known Marie Unna locus at 8p21 near the Hairless gene were performed. Linkage analysis using markers D8S298, D8S560, D8S258, and D8S282 revealed significant exclusion of this locus (Z = -2.0 or lower) at theta = 0.1. This demonstrates that this novel ectodermal dysplasia is both phenotypically and genetically distinct from Marie Unna hereditary hypotrichosis. PMID- 12895009 TI - The depression gender gap: a view through a biocultural filter. AB - That women, compared with men, are more prone to become "depressed" has been known for decades. The etiology of the depression gender gap has been a source of much discussion, with the bulk occurring within the medical model. As a complement to--rather than a competitor with--such discussion, the depression gender gap is analyzed through a biocultural lens wherein the evolutionary history of Homo sapiens becomes part of the frame of reference. Given that a "constant" is difficult to explain by the use of "variables," the very consistent 2:1 gender ratio of female to male (rates of) depression is difficult to explain by referring to cultural parameters, which are variable. It is more parsimonious to suggest that a constant becomes a better candidate to explain another constant. The constant used in this exercise is some portion of the genetic package that subtends the species Homo sapiens; in other words, everyone on the planet is human. The authors argue that the basal dynamic of H. Fisher's (1983) sex contract is a useful analytical tool in examining the contemporary gender gap in depression. They suggest that the germane clause of the sex contract is the use of psychological immobilization as an effective social instrument. PMID- 12895010 TI - Individual differences in infant fearfulness and cognitive performance: a testing, performance, or competence effect? AB - The author conducted 2 studies to examine the relations between infant fear and cognitive testing performance in 12-month-old infants. In Study 1, fear was assessed by using 2 standard temperament questionnaires and a laboratory-based, standardized stranger approach. Individual differences in cognitive development were assessed using the Object Permanence Scale of the Infant Psychological Development Scales (I. C. Uzgiris & J. M. Hunt, 1975). All 3 assessments of fear significantly predicted object permanence performance, with correlations ranging from -.32 to -.35. In Study 2, fear was assessed via a maternal report questionnaire, and habituation performance was assessed via a basic-level categorization task. Familiarity with the examiner and with the testing environment was manipulated to test for a familiarity influence on performance. Testing revealed individual differences in both fear and habituation. Results suggest that highly fearful infants required more trials to habituate and were less likely to meet the habituation criterion than infants who were less fearful. Methodological and conceptual implications of these results are discussed. PMID- 12895011 TI - The impact of father absence on adolescent separation-individuation. AB - The authors compared separation-individuation and psychological separation from fathers of 25 adolescent boys who were living with both biological parents with that of 25 boys who were living with their biological mothers in homes in which the fathers did not reside. The results showed that the boys in the 2 groups did not differ on measures of separation-individuation and that the quality of the mother-son relationship mediated several of the assessed separation-individuation manifestations. The authors' initial data analysis with regard to psychological separation showed that boys who lived in homes in which the father was a nonresident were more separated on 2 of the 4 dimensions assessed; however, when controlled for quality of mother-son and father-son relationship, these differences were not significant. The frequency of father contact in homes in which the father did not reside was positively correlated with healthy separation but negatively correlated with functional, attitudinal, and emotional independence from the father. The results of the study support the importance of the quality of a child's relationship with his or her mother and father as a mediator of several dimensions of the separation-individuation process. PMID- 12895012 TI - Posttraumatic stress symptoms in adolescent survivors of childhood cancer and their mothers. AB - We examined symptoms of posttraumatic stress disorder (PTSD) in adolescent survivors of childhood cancer and their mothers and the contribution of family functioning, including perceived emotional support and familial conflict, and individual factors including life stress and severity of disease to PTSD symptoms. Participants were 52 adolescent cancer survivors and their mothers and 42 healthy adolescent counterparts and their mothers. Findings revealed that mothers of cancer survivors endorsed more PTSD symptoms than did their healthy counterparts and that survivors and mothers also reported greater recent and past stressful life events. Although no survivors met clinical criteria for a PTSD diagnosis, over 36% endorsed mild subthreshold symptomatology. Findings are discussed in the context of understanding PTSD symptoms within a family systems framework. PMID- 12895013 TI - Hypothalamic-pituitary-adrenal activity among Armenian adolescents with PTSD symptoms. AB - This study evaluated basal levels and responsiveness to exercise of plasma adrenocorticotropic hormone (ACTH), and serum thyroid stimulating hormone (TSH), growth hormone (GH) and cortisol among adolescents from two differentially exposed groups 6 1/2 years after the 1988 earthquake in Armenia. Severity of total PTSD and Category C and D symptoms were negatively correlated with baseline cortisol. Preexercise ACTH was significantly lower, and preexercise TSH higher, among adolescents with more exposure. Depressive symptoms were negatively correlated with baseline cortisol and positively with TSH. Mean GH, TSH, and cortisol levels in both groups fell within normal limits. The pre- to postexercise increase in GH, TSH, and cortisol suggests that exercise challenge may be useful in the field investigation of neurohormonal activity among traumatized individuals. PMID- 12895014 TI - The relationship among cognitive schemas, job-related traumatic exposure, and posttraumatic stress disorder in journalists. AB - American newspaper journalists (N = 906) participated in a study examining a cognitive mediational model for explaining the relationship between exposure to work-related traumatic events and work-related posttraumatic stress disorder (PTSD). Results indicated (a) greater exposure to work-related traumatic events was associated with work-related PTSD symptoms, as well as negative cognitive schemas; (b) cognitive beliefs partially accounted for PTSD symptoms, but the full cognitive mediational model was not supported. Implications include targeting interventions for journalists who experience traumatic stress and modifying theories about PTSD symptoms in journalists. PMID- 12895015 TI - Ambulatory monitoring and physical health report in Vietnam veterans with and without chronic posttraumatic stress disorder. AB - This study investigated the associations among PTSD, ambulatory cardiovascular monitoring, and physical health self-reports in 117 male Vietnam combat veterans (61 with PTSD and 56 without PTSD). PTSD was associated with health symptoms and number of current health conditions beyond the influence of several covariates. PTSD was associated with greater systolic blood pressure variability, and an elevated percentage of heart rate and systolic blood pressure readings above baseline. Higher mean heart rate and an elevated percentage of heart rate above baseline were associated with physical health symptoms. None of the ambulatory monitoring variables mediated the association between PTSD and physical health outcomes. Findings suggest that the interrelationships among ambulatory autonomic responses, PTSD, and physical health deserve more research attention. PMID- 12895016 TI - Group therapy for incarcerated women who experienced interpersonal violence: a pilot study. AB - This study evaluated effectiveness of group therapy for incarcerated women with histories of childhood sexual and/or physical abuse. The intervention was based on a two-stage model of trauma treatment and included Dialectical Behavior Therapy skills and writing assignments. We randomly assigned 24 participants to group treatment (13 completed) and 25 to a no-contact comparison condition (18 completed). We evaluated treatment effects, using the Beck Depression Inventory, Inventory of Interpersonal Problems, and Trauma Symptom Inventory. The data demonstrate significant reductions in PTSD, mood, and interpersonal symptoms in the treatment group. PMID- 12895017 TI - Are the dissociative criteria in ASD useful? AB - Acute stress disorder (ASD) is a new DSM-IV diagnostic category, characterized by dissociative, intrusive, avoidance, and hyperarousal symptoms in the first month after a traumatic experience. The goal of the present study was to examine the utility of this diagnosis. In a prospective study, 79 mixed trauma victims who met DSM-IV symptom criteria for PTSD within 1 month following a traumatic event were followed through three months post-event. Dissociative symptoms in ASD only partially captured distress and dysfunction during the first month. Participants with and without ASD showed similar patterns of recovery, with only small differences that disappeared at three months post-event. Interestingly, initial PTSD avoidance but not ASD dissociative symptoms predicted PTSD severity at 3 months. PMID- 12895018 TI - Mental health, social functioning, and feelings of hatred and revenge of Kosovar Albanians one year after the war in Kosovo. AB - A cross-sectional cluster sample survey was conducted in June 2000 in Kosovo to assess the prevalence of mental health problems associated with traumatic experiences, feelings of hatred and revenge, and the level of social functioning among Kosovar Albanians approximately 1 year after the end of the war. Findings of the second cross-sectional survey were compared with those from our 1999 mental health survey in Kosovo. Included in the survey were 1399 Kosovar Albanians aged 15 years or older living in 593 randomly selected households across Kosovo. Twenty-five percent of respondents reported PTSD symptoms, compared with 17.1% in 1999. The MOS-20 social functioning score improved to 69.8 from 29.5 in 1999. In the 2000 survey 54% of men felt hatred toward the Serbs, compared with 88.7% in 1999. PMID- 12895019 TI - Predictors of chronic posttraumatic stress symptoms following burn injury: results of a longitudinal study. AB - The authors' goal was to examine the course and predictors of posttraumatic stress symptoms among persons hospitalized for burns. A total of 301 participants completed self-report measures assessing peritraumatic mental state, anxiety related to pain, and posttraumatic stress symptoms. Twenty-six percent of the participants were suffering from posttraumatic stress symptoms at 2-3 weeks postburn and 15% of them at 12 months postburns. In general, a decrease in symptoms was observed over time, although a substantial part of the participants with acute stress symptoms suffers from chronic posttraumatic stress symptoms 1 year postburn. Symptoms were predicted by anxiety measures and objective factors, such as female gender, locus, and severity of injury. PMID- 12895020 TI - The relationship between intimate partner violence and PTSD: an application of Cox regression with time-varying covariates. AB - This study uses Cox regression with time-varying covariates to examine the relationship between intimate partner violence and posttraumatic stress disorder (PTSD) in a random sample of Japanese American women and immigrant women from Japan (N = 211). Because applications of survival analysis in trauma research are scarce, this paper presents the utility of this analytical approach by contrasting it with other common methods of analysis (chi-square tests and Cox regression with covariates that do not change over time). PMID- 12895021 TI - Male-perpetrated violence among Vietnam veteran couples: relationships with veteran's early life characteristics, trauma history, and PTSD symptomatology. AB - Using structural equation modeling, we examined the impact of early-life stressors, war-zone stressors, and PTSD symptom severity on partner's reports of recent male-perpetrated intimate partner violence (IPV) among 376 Vietnam veteran couples. Results indicated that several variables demonstrated direct relationships with IPV, including relationship quality with mother, war-zone stressor variables, and PTSD symptom severity. Importantly, retrospective reports of a stressful early family life, childhood antisocial behavior, and war-zone stressors were indirectly associated with IPV via PTSD. One of our 2 war-zone stressor variables, perceived threat, had both direct and indirect (through PTSD) relationships with IPV. Experiencing PTSD symptoms as a result of previous trauma appears to increase an individual's risk for perpetrating IPV. Implications for research and treatment are discussed. PMID- 12895022 TI - Changes in religious beliefs following trauma. AB - Information processing theorists propose that traumatic events can lead to disruptions in the processing of information and to changes in beliefs. This study examined the relationships among trauma, posttraumatic stress disorder (PTSD), and religious beliefs. Participants included 120 individuals from community and clinical samples who participated in the DSM-IV Field Trial Study on PTSD. Results indicated that the PTSD group was more likely to report changes in religious beliefs following the first/only traumatic event, generally becoming less religious. PTSD status was not related to change in religious beliefs following the most recent event. Intrinsic religiosity was related to multiple victimization, but not PTSD. Results are discussed in terms of understanding the function of religiosity in participants' lives and future directions for research. PMID- 12895023 TI - Consistency of retrospective reporting about exposure to traumatic events. AB - Lifetime exposure to traumatic events was assessed by means of a multimethod protocol applied to 76 male military veterans. Consistency of retrospective reporting was determined for physical and sexual assault and abuse, accidents, disasters, combat and warzone experiences, serious illness or injury, and hazardous duty. Findings demonstrate that respondents are generally consistent in reporting traumatic events, although the majority report more events upon reevaluation. Reporting about traumatic events shows some variation as a function of the life epoch in which events occurred, whether they were directly or indirectly experienced, and the type of trauma involved. Discussion addresses memory-related processes triggered by trauma evaluation or tied to characteristics of events themselves as potential sources of inconsistency. PMID- 12895024 TI - "Forward psychiatry" in the military: its origins and effectiveness. AB - "Forward psychiatry" was devised in World War I for the treatment of shell shock and today is the standard intervention for combat stress reaction. It relied on three principles: proximity to battle, immediacy, and expectation of recovery, subsequently given the acronym "PIE." Both US and UK forces belatedly reintroduced PIE methods during World War II to return servicemen to active duty and made confident claims for its efficacy. Advanced treatment units also appeared to have minimized psychiatric battle casualties during Korean and Vietnamese Wars. Evaluations of its use by Israeli forces in the Lebanon conflict showed higher return-to-duty rates than at base hospitals. A reexamination of these examples suggests that reported outcomes tended to exaggerate its effectiveness both as a treatment for acute stress reaction and as a prophylaxis for chronic disorders such as PTSD. It remains uncertain who is being served by the intervention: whether it is the individual soldier or the needs of the military. PMID- 12895025 TI - Gender, social support, and PTSD in victims of violent crime. AB - Gender differences in social support levels and benefits were investigated in 118 male and 39 female victims of violent crime assessed for PTSD symptomatology 1 and 6 months postcrime. Within 1 month postcrime both genders reported similar levels of positive support and support satisfaction, but women reported significantly more negative responses from family and friends. Women also reported an excess of PTSD symptoms that was similar at 1 and 6 months postcrime, and negative responses mediated the relationship between gender and later symptoms. Overall negative response and support satisfaction, but not positive support, were significantly associated with PTSD symptoms. However, the effects of support satisfaction and negative response on 6-month symptoms were significantly greater for women than men. The findings are consistent with previous studies of predominantly female assault victims concerning the stronger impact of negative over positive support, and might help explain women's higher PTSD risk in civilian samples. PMID- 12895026 TI - Involvement of endothelin receptors in normal and pathological development of neural crest cells. AB - Endothelin receptors (Ednr) are G-protein-coupled receptors with seven membrane spanning domains and are involved in various physiological processes in adults. We review here the function of these receptors during the development and transformation of the neural crest cell-specific lineage. Neural crest cells (NCC) may be classified according to their location in the body. In particular, there are clear differences between the neural crest cells arising from the cephalic part of the embryo and those arising from the vagal and truncal part. The development of cranial and cardiac NCC requires the endothelin-1/Ednra system to be fully functional whereas the development of more posterior NCC requires full functionality of the endothelin-3/Ednrb system. Mutations have been found in the genes corresponding to these systems in mammals. These mutations principally impair pigmentation and enteric ganglia development. The precise patterns of expression of these receptors and their ligands have been determined in avian and mammalian models. Data obtained in vitro and in vivo have provided insight into the roles of these proteins in cell proliferation, migration, differentiation and transformation. PMID- 12895027 TI - Ca2+-independent protein kinase C signalling in mouse eggs during the early phases of fertilization. AB - Protein kinase C (PKC), an enzyme playing a central role in signal transduction pathways, is activated in fertilized mouse eggs downstream of the fertilization Ca2+ signal, to regulate different aspects of egg activation. Given the presence of Ca2+-independent PKC isoforms within the egg, we investigated whether fertilization triggers PKC stimulation in mouse eggs by activating Ca2+ independent signalling pathways. An increase in PKC activity was detected as early as 10 min after the beginning of insemination, when about 90% of eggs had fused with sperm and the first Ca2+ rise was evident in most of the eggs. A similar level of activity was found 20 min later, when about 60% of eggs had resumed meiosis. When the Ca2+ increase was buffered by an intracellular Ca2+ chelating agent, PKC stimulation was not blocked but only slightly reduced. Confocal microscopy analysis revealed that the increase in PKC activity at fertilization coincided with the translocation of PKCdelta, a Ca2+-independent and diacylglycerol-dependent PKC isoform, to the meiotic spindle. When, in the absence of the Ca2+ signal, metaphase-anaphase transition was inhibited, PKCdelta moved to the meiotic spindle but still maintained a sustained cytoplasmic distribution. In summary, our results indicate that: 1) PKC activation is an early event of egg activation; 2) both Ca2+-dependent and Ca2+-independent pathways contribute to increased PKC activity at fertilization; 3) PKCdelta is one of the isoforms participating in this signalling process. PMID- 12895028 TI - Dlx5 regulates chondrocyte differentiation at multiple stages. AB - Endochondral ossification, in which cartilaginous templates are progressively replaced by marrow and bone, represents the dominant mode of development of the axial and appendicular skeleton of vertebrates. Chondrocyte differentiation within the cartilaginous core of these skeletal elements is tightly regulated, both spatially and temporally. Here, we describe the expression of Dlx5 in the cartilaginous core of limb skeletal elements in chicken and mouse embryos. We find that Dlx5 is one of the earliest genes expressed in condensing limb mesenchyme that will give rise to the limb skeleton. Later, when proliferating and differentiating chondrocytes are found in spatially distinct regions of the cartilaginous model, Dlx5 is expressed in the zone of hypertrophy and in proliferating chondrocytes that are poised to differentiate. Consistent with this pattern of expression, we show that forced expression of Dlx5 potentiates early and late chondrocyte differentiation and inhibits proliferation in cultured cells. Examination of the limbs of mutant Dlx5 mouse embryos revealed that they displayed a delay in chondrocyte maturation compared with wild type littermates. Taken together, our data reveal a positive role for Dlx5 during multiple stages of chondrocyte differentiation and, along with previous studies of Dlx5 and osteogenesis, identify Dlx5 as a general regulator of differentiation in the mouse skeleton. PMID- 12895029 TI - Receptor protein tyrosine phosphatase mu expression as a marker for endothelial cell heterogeneity; analysis of RPTPmu gene expression using LacZ knock-in mice. AB - The receptor-like protein tyrosine phosphatase mu (RPTPmu) belongs to the subfamily of meprin, A5, RPTPmu (MAM) domain-containing RPTPs, which are thought to play an important role in cell-cell adhesion mediated processes. The current study was designed to examine the expression pattern of RPTPmu in mice. We have generated RPTPmu-LacZ knock-in mice that express the beta-galactosidase (LacZ) reporter gene under the control of the RPTPmu promoter. LacZ expression patterns were analysed in embryos and adult mice by whole mount LacZ staining. Analysis of beta-galactosidase activity of heterozygous embryos and adult tissues revealed RPTPmu expression in endothelial cells of arteries and capillaries. In contrast, expression was virtually absent in endothelial cells of veins and in fenestrated endothelial cells in the adult liver and spleen. Moreover, RPTPmu expression was found in endothelial cells from the endocardium and the aorta in embryos, but not in adult mice. In addition to heterogeneous expression in endothelial cells, RPTPmu expression was found in cardiac muscle cells but not in skeletal muscle cells or smooth muscle cells. Expression was also found in Type II pneumonocytes in the lung alveoli and in Purkinje cells and other neurons in the brain. The specific expression of RPTPmu in arterial endothelial cells and in cardiac myocytes suggests that RPTPmu may play a role in the regulation of cardiovascular functions. PMID- 12895030 TI - The proliferative state, graft-site and contact-time of competent chick ectoblast determine the quality and quantity of neural induction by Hensen's node. AB - We have assessed the type and amount of neural tissue induced in the chick gastrula ectoblast with increasing duration of contact with the Hensen node inducer. At least 4 h contact is necessary to induce the ectoblast in the area pellucida, 9 h in the area opaca and even longer at the margin of overgrowth. In the area pellucida, the inductive response shifts from archencephalic type at 4-6 h contact to deuterencephalic type after 9 h contact. The induced tissue volume and cell number increased as graft-host contact increased from 4-9 h, and then decreased with longer contact. We suggest that in addition to the period of contact and the grafting position on competent ectoblast, the rate of cell proliferation may control the axial specificity and morphological organization of the induced neural tissue. PMID- 12895031 TI - Abnormal development of the diaphragm in mdx:MyoD-/-(9th) embryos leads to pulmonary hypoplasia. AB - In vitro studies have shown that mechanical factors play an important role in cell cycle kinetics and cell differentiation of the lung through an unknown mechanochemical signal transduction pathway. In this study we evaluated the in vivo role of mechanical factors due to fetal breathing movements (primarily executed by the diaphragm, which is the main respiratory muscle) on lung growth and development by using genetically engineered embryos. Lung growth and development of wild-type, mdx:MyoD+/-(9th) (in which the diaphragm develops normally) and mdx:MyoD-/-(9th) (in which the diaphragm muscle is significantly thinned and not functional) embryos were compared at embryonic day 18.5 using immunohistochemistry, in vivo TUNEL detection and morphometry. No abnormalities in lung organogenesis were observed in mdx:MyoD+/-(9th) term embryos, whereas lung hypoplasia was detected in mdx:MyoD-/-(9th) embryos. In the hypoplastic lung, the number of proliferating lung cells was lower in comparison to the wild type and mdx:MyoD+/-(9th) embryos, while the gradient of thyroid transcription factor-1 (TTF-1) was not maintained. Surprisingly, no difference was observed in distribution and occurrence of apoptotic lung cells in mdx:MyoD-/-(9th) embryos. Together, it appears that mechanical forces generated by contractile activity of the diaphragm muscle play an important role in normal lung growth and development by affecting cell proliferation and TTF-1 expression. PMID- 12895032 TI - Immunocytochemistry of the amphibian embryo--from overview to ultrastructure. AB - Amphibian embryos are standard research objects to study pattern formation and morphogenesis. Due to their external development and robust nature, experimental manipulations such as microinjections or transplantations can be easily performed. However, most immunocytochemical approaches addressing the specific localization of proteins are hampered by the fragility of the large and yolky embryonic cells which render high resolution staining difficult. Immunocytochemical data are therefore often restricted to either overall patterns in whole embryo preparations or to immunofluorescent localization with limited resolution on sections. High resolution or ultrastructural protein localization data are rare and can be achieved only with time consuming procedures. Here, a comparative study of immunocytochemical methods suitable for light and electron microscopy using different kinds of plastic resins is presented. Three main approaches are described: preembedding staining of whole embryos, postembedding staining of ultrathin sections and preembedding staining of vibratome sections. All the procedures are designed to study protein expression in early amphibian embryos en gros as well as en detail and the described techniques are suitable to combine two or three levels of resolution on the very same biological specimen. Examples are presented and advantages and disadvantages of the different protocols are discussed. PMID- 12895033 TI - Ventral nerve cord remodeling in a stingless bee (Melipona quadrifasciata anthidioides, Hymenoptera, Apidae) depends on ecdysteroid fluctuation and programmed cell death. AB - The reorganization of the ventral nerve cord (VNC) during metamorphosis of M. quadrifasciata was observed to be characterized by shortening of connectives and subsequent fusion of the 2nd and 3rd thoracic and the 1st abdominal ganglia. Also, the 5th to 7th abdominal ganglia came into very close contact. These changes were accompanied by increasing levels of endogenous ecdysteroids, as determined by a radioimmunoassay. Incubation of VNC in the presence of 5 microg 20-hydroxyecdysone, caused significant shortening of connectives in the thoracic region, but not in the abdomen, evidencing a segment-specific response to this hormone. Cell death in the ventral ganglia was revealed by transmission electron microscopy and TUNEL-reaction. Detection of labeled cells in the region where contiguous ganglia come into close contact suggests that programmed cell death is involved in ganglionic fusion. PMID- 12895034 TI - Bridged triarylamines: a new class of heterohelicenes. AB - A series of bridged triarylamines, which constitute a new class of heterohelicenes, have been synthesized using a simple three-step procedure. These compounds are shown to be highly luminescent chromophores and are capable of being oxidized. The solid-state structures of these helicenes show a tendency for pi-stacking interactions into an overall zigzag motif. PMID- 12895035 TI - Cytosporacin, a highly unsaturated polyketide: application of the ACCORD-ADEQUATE experiment to the structural determination of natural products. AB - Cytosporacin (1), a novel antibacterial polyketide containing naphthopyranone and isochromandione moieties, was isolated from the fermentation broth of the fungus Cytospora rhizophorae. A (1)H-detected ACCORD-ADEQUATE pulse sequence that distinguished (2)J(CH) from (3)J(CH) correlations provided critical information for structural determination. NOE studies established the relative configuration and revealed the presence of two rotamers. A biosynthetic (13)C-labeling experiment indicated that cytosporacin was derived from acetate origin. PMID- 12895036 TI - Methyl and silyl mesolytic dissociations in the radical cations and radical anions of but-1-ene, allylsilane, hexa-1,3-diene, and penta-2,4-dienylsilane. CAS MCSCF and coupled cluster theoretical study. AB - Methyl or silyl dissociation in the CH(2)=CHCH(2)-XH(3) (a-XH(3)(*)(+)) and CH(2)=CHCH=CHCH(2)-XH(3) (p-XH(3)(*) (+)) radical cations (X = C, Si) yields a(+) or p(+) and XH(3)(*). Similarly, the radical anions a-CH(3)(*) (-) and p-CH(3)(*) (-) give the pi-delocalized anion and CH(3)(*) preferentially. In contrast, a SiH(3)(*) (-) and p-SiH(3)(*-) prefer to dissociate into the pi-delocalized radical and silide. All reactions are endoergic: by 43-50 kcal mol(-)(1) in the radical cations, and easier to some extent in the radical anions, that require 29 33 (X = C) and 13-14 kcal mol(-)(1) (X = Si). The fragmentation energy profiles do not present significant barriers for the backward process in the case of the radical cations. All radical anions exhibit an energy maximum along the dissociation pathway, but the barrier is lower than the dissociation limit. Fragmentation is "activated" more in the anions than in the cations with respect to homolysis in the corresponding neutrals (that requires 72-81 kcal mol(-)(1)). Wave function analysis indicates that the C-X bond cleavage in the hydrocarbon radical ions, although formally comparable to a homolytic process, is at variance with this model, due to the spin recoupling of one of the two C-X bond electrons with the originally unpaired electron. This is basically true also for the silyl substituted radical anions, in which the initial more delocalized charge distribution might suggest some heterolytic character of the bond cleavage. PMID- 12895037 TI - Convergent enantioselective synthesis of vinigrol, an architecturally novel diterpenoid with potent platelet aggregation inhibitory and antihypertensive properties. 1. Application of anionic sigmatropy to construction of the octalin substructure. AB - The coupling of building blocks 15 and 36e in the presence of MgBr(2).OEt(2) at 0 degrees C proceeds with an exo stereoselectivity (3.2:1) considerably more advantageous for the acquisition of carbinol 37e than in the absence of the additive (exo/endo = 1:5.7). The pivotal transformation that sets all of the relevant stereocenters of the cis-octalin 55 is the oxyanionic-accelerated [3,3] sigmatropic rearrangement of 37e. A salient feature is the structurally enforced adoption of a boatlike transition state that serves to properly set four vicinal methine hydrogens in an all-cis arrangement. The ensuing conversion of 55 into iodo sulfone 62 has permitted X-ray crystallographic confirmation of all absolute stereochemical assignments since the isopropyl substituent was initially installed enantioselectively via the Evans oxazolidinone protocol. No intramolecular anionic cyclization of 62 to generate the tricyclic framework was seen. This absence of reactivity is attributed to conformational factors that inhibit attainment of the proper S(N)2 reaction trajectory. PMID- 12895038 TI - Synthesis of 5'-amino-5'-phosphonate analogues of pyrimidine nucleoside monophosphates. AB - The 5'-amino-5'-phosphono derivatives of cytidine, cytosine arabinoside (ara-C), and uridine have been prepared via the corresponding nucleoside aldehydes. Phosphite addition to imines derived from the nucleoside aldehydes and p methoxybenzylamine was efficient, and use of this amine allowed cleavage of the products to the parent amino phosphonic acids. The phosphite additions proved to be diastereoselective, with the cytidine and uridine derivatives favoring the 5'S stereochemistry and the ara-C derivative favoring the 5'R isomer. The stereochemistry of one cytidine derivative was established by single-crystal diffraction analysis, and detailed comparisons of the (13)C NMR data allowed assignments of the other amino phosphonates. PMID- 12895039 TI - Novel photoswitchable receptors: synthesis and cation-induced self-assembly into dimeric complexes leading to stereospecific [2+2]-photocycloaddition of styryl dyes containing a 15-crown-5 ether unit. AB - Styryl dyes 4a-e containing a 15-crown-5 ether unit and a quinoline residue with a sulfonatoalkyl or sulfonatobenzyl N-substituent were synthesized. The relationship between the photochemical behavior of these dyes and their aggregates derived from complexation with Mg(2+) in MeCN was studied using (1)H NMR and absorption spectroscopy. The E-isomers of 4a-e were shown to form highly stable dimeric (2:2) complexes with Mg(2+). Upon irradiation with visible light, the dimeric complexes undergo two competing photoreactions, viz., geometric E --> Z isomerization, resulting in an anion-capped 1:1 complex of the Z-isomer with Mg(2+) and stereospecific syn-head-to-tail [2+2]-cycloaddition, affording a single isomer of bis-crown-containing cyclobutane. The N-substituent in the dye has a dramatic effect on the photochemical behavior of the dimeric complex. Molecular dynamics and semiempirical quantum-chemical calculations were carried out to interpret the observed photocycloaddition in the dimer. Conformational equilibria for the dimer of (E)-4b were analyzed using (1)H NMR spectroscopy. PMID- 12895040 TI - New strategic reactions for organic synthesis: catalytic asymmetric C-H activation alpha to oxygen as a surrogate to the aldol reaction. AB - The C-H activation of silyl ethers by means of rhodium carbenoid-induced C-H insertion represents a very direct method for the stereoselective synthesis of silyl-protected beta-hydroxy esters. The reaction can proceed with very high regio-, diastereo-, and enantioselectivity and represents a surrogate to the aldol reaction. The reaction is catalyzed by the rhodium prolinate complex Rh(2)(S-DOSP)(4). A critical requirement for the high chemoselectivity is the use of donor/acceptor-substituted carbenoids such as those derived from methyl aryldiazoacetates. A range of silyl ethers may be used such as allyl silyl ethers, tetraalkoxysilanes, and even simple trimethylsilyl alkyl ethers. In general, C-H activation preferentially occurs at methylene sites, as the reactivity is controlled by a delicate balance between steric and electronic effects. PMID- 12895041 TI - Application of vicarious nucleophilic substitution to the total synthesis of dl physostigmine. AB - A concise, highly efficient formal total synthesis of dl-physostigmine is described, using a relatively simple method that should be adaptable to the synthesis of homologous members of this type of alkaloid. The key step in the synthesis is a new vicarious nucleophilic substitution reaction between p nitroanisole and a C-silylated derivative of N-methylpyrrolidinone. Subsequent conversion of the initial adduct to the tricyclic framework of physostigmine follows a well-established protocol and provides the key intermediate 8 in high yield. The vicarious nucleophilic substitution reaction has also been extended to six-membered lactams, with encouraging results. PMID- 12895042 TI - Cascade polycyclization: exploration of a convergent route to access various tricyclic and tetracyclic products related to sterols. AB - The expedient synthesis of tricyclic and tetracyclic compounds via a cascade polycyclization methodology is described. Nazarov substrates (II) containing two Michael acceptors and a cyclohexenone ester (I) underwent cycloaddition followed by intramolecular 1,4-addition to furnish, in a highly stereoselective manner, tricyclic and tetracyclic products (III). Such compounds are interesting intermediates for the synthesis of polycyclic natural and unnatural products. PMID- 12895043 TI - Pd(II)-catalyzed coupling cyclization of 2,3-allenoic acids with allylic halides. An efficient methodology for the synthesis of beta-allylic butenolides. AB - An effective method for the synthesis of beta-allyl polysubstituted butenolides from the easily available allylic halides and 2,3-allenoic acids is described. By using this method optically active butenolides can be obtained. According to the results presented in this paper, the reaction may proceed via three consecutive steps: cyclic oxypalladation of the allene, insertion of the C=C bond in allylic halides, and beta-dehalopalladation. PMID- 12895044 TI - The scope of catalytic enantioselective tandem carbonyl ylide formation intramolecular [3 + 2] cycloadditions. AB - Catalytic enantioselective tandem carbonyl ylide formation-intramolecular 1,3 dipolar cycloaddition reactions of 2-diazo-3,6-diketoesters show promising scope in terms of asymmetric induction as the tethered alkene/alkyne dipolarophile component is varied. Cycloadditions were found to occur in moderate to very good yields, with a difference in ee exhibited by the electronically different 2-diazo 3,6-diketoesters 1, 25 and 33, 34. Values for ee of up to 90% for alkene dipolarophiles and up to 86% for alkyne dipolarophiles were obtained. PMID- 12895045 TI - Total synthesis of (+/-)-kainic Acid with an aza-[2,3]-Wittig sigmatropic rearrangement as the key stereochemical determining step. AB - A flexible route to the kainoid skeleton is exemplified by the synthesis of (+/-) kainic acid from 3-butyn-1-ol. The route relies on the aza-[2,3]-Wittig sigmatropic rearrangement to efficiently install the relative stereochemistry between C2-C3. The C4 stereocenter was derived from a diastereocontrolled iodolactonization. The aza-[2,3]-Wittig rearrangement potentially allows structural diversity at C3 and the displacement of the tosyloxy group with retention of stereochemistry allows structural diversity at C4. The trans-C2 carboxylic acid functional group was found to be the most important for retention of stereochemistry at C4 upon treatment with a higher order cyano cuprate reagent. PMID- 12895046 TI - 1,4-silatropy of S-alpha-silylbenzyl thioesters: a convenient route to silyl enol and dienol ethers accompanied by C-C bond formation via thiocarbonyl ylides. AB - A novel convenient method for the generation of thiocarbonyl ylides from readily accessible starting materials and the first synthetic application of in situ generated ylides in the synthesis of silyl enol and dienol ethers, accompanied by C-C bond formation, is described. Under completely neutral conditions without any catalyst or additive, thermal reactions of S-alpha-silylbenzyl thioesters in sealed tubes at 180 degrees C provided silyl enol and dienol ethers in good to excellent yields with high stereoselectivities. This procedure consists of a multistep reaction in a one-pot process, i.e., 1,4-silatropy of S-alpha silylbenzyl thioesters to give thiocarbonyl ylides, 1,3-electrocyclization of the ylides to give thiiranes, and the extrusion of sulfur from thiiranes to give silyl enol and dienol ethers. PMID- 12895047 TI - Model studies toward the synthesis of dihydropyrimidinyl and pyridyl alpha-amino acids via three-component Biginelli and Hantzsch cyclocondensations. AB - A novel and versatile strategy for the synthesis of heterocyclic alpha-amino acids has been described. The use of components (aldehyde or beta-ketoester) bearing a masked glycinyl moiety in Biginelli and Hantzsch cyclocondensations allowed access to the 4-dihydropyrimidinyl-alpha-glycines, 4-dihydropyrimidinyl alpha-alanines, 4-pyridyl-alpha-alanines, and 2-pyridyl-alpha-alanines classes. Dihydropyrimidinyl-amino acids were obtained as a mixture of diastereoisomers due to the formation of the stereocenter at C4 of the dihydropyrimidinone ring. Individual stereoisomers were isolated as pure compounds and their structures were assigned with the aid of X-ray crystallography and chiroptical properties. The enantiomeric purity of a representative selection of the above amino acids was greater than 96% as verified by derivatization to the corresponding Mosher's amides and subsequent (1)H and (19)F NMR spectroscopy. Incorporation of the 4 pyridyl-alpha-alanine derivative into a peptide chain is also described. PMID- 12895048 TI - Acyclic congener of cucurbituril: synthesis and recognition properties. AB - The cucurbit[n]uril (CB[n]) family of macrocycles occupies a prominent role in molecular recognition and self-assembly studies despite the current inability to access specific cucurbit[n]uril homologues, derivatives, and analogues by straightforward tailor-made synthetic procedures. In this paper, we explore an approach that circumvents the challenges posed by the tailor-made synthesis of macrocyclic CB[n] by preparing 1, which functions as an acyclic CB[6] congener. The o-xylylene connections to the glycoluril rings preorganize 1 into the (a,a,a,a)-1 conformation required for binding and reduce its tendency to undergo self-association. We surveyed the binding properties of 1 toward 16 amines (K(a) C3-O(i)()(-)(1) and a syn orientation between the C2-H and CO. In this molecule, the C2-hydrogens and the COs can be seen on one side of the ring while the NHs point to the other side. Addition of the bicyclic lactam 5b resulted in the influx of Na(+) ions across the lipid bilayer leading to the dissipation of valinomycin-mediated K(+) diffusion potential. PMID- 12895059 TI - Highly diastereoselective strecker reaction of enolizable aliphatic sulfinimines. AB - The reaction of chiral sulfinimines 1c-g derived from aliphatic aldehydes with TMSCN in the presence of CsF gave alpha-amino nitriles in high diastereoselectivity and yield. alpha,beta-Diamino acid derivatives were also obtained in high diastereoselectivity from the reaction of 2 aziridinesulfinimines 1h and 1i followed by ring-opening of the products with thiophenol. The presence of hydrogen at the alpha-position of the C=N double bond is crucial in this TMSCN addition reaction. PMID- 12895060 TI - Stereoselective synthesis of the nonproteinogenic amino acid (2S,3R)-3-amino-2 hydroxydecanoic acid from (4S,5S)-4-formyl-5-vinyl-2-oxazolidinone. AB - (4S,5S)-4-Formyl-5-vinyl-2-oxazolidinone (4b), which is readily obtained via a zinc-silver-mediated reductive elimination of alpha-d-lyxofuranosyl phenyl sulfone (3b), is successfully converted to the naturally occurring, nonproteinogenic amino acid (2S,3R)-3-amino-2-hydroxydecanoic acid (2). Also in this study, a facile "oxazolidinone rearrangement" reaction is uncovered during the attempted formation of the (methylthio)thiocarbonate derivative of the oxazolidinone alcohol 7. PMID- 12895061 TI - Absolute structures of some naturally occurring isopropenyldihydrobenzofurans, remirol, remiridiol, angenomalin, and isoangenomalin. AB - The absolute structures of some naturally occurring chiral 2-isopropenyl-2,3 dihydrobenzofurans, (+)-remirol (1a), (+)-remiridiol (1b), (+)-angenomalin (2), and (+)-isoangenomalin (3), were studied by respective chiral synthesis. Kinetic resolutions of racemic 2-isopropenyl-2,3-dihydrobenzofurans, 2-isopropenyl-4,6 dimethoxy-2,3-dihydrobenzofuran (4), 4-hydroxy-2-isopropenyl-2,3 dihydrobenzofuran-5-carbaldehyde (8), and 2-isopropenyl-6-(MOM)oxy-2,3 dihydrobenzofuran-5-carbaldehyde (11c), by Sharpless dihydroxylation using (DHQ)(2)AQN or (DHQD)(2)AQN gave the corresponding chiral 2-isopropenyl-2,3 dihydrobenzofurans. Chiral (S)-(+)-4 (99% ee, using (DHQD)(2)AQN) was converted to natural remirol (S)-(+)-1a and then to natural remiridiol (S)-(+)-1b. (S)-(+) 8 (97% ee, using (DHQD)(2)AQN) was converted to natural angenomalin (S)-(+)-2. (R)-(-)-11c (>99% ee, using (DHQ)(2)AQN), was converted to natural isoangenomalin (R)-(+)-3. Thus, the absolute structures of natural remirol (+)-1a and remiridiol (+)-1b and angenomalin (+)-2 were determined to be S, and the structure of natural isoangenomalin (+)-3 was R. PMID- 12895062 TI - The enantiospecific, stereospecific total synthesis of the ring-A oxygenated sarpagine indole alkaloids (+)-majvinine, (+)-10-methoxyaffinisine, and (+)-N(a) methylsarpagine, as well as the total synthesis of the alstonia bisindole alkaloid macralstonidine. AB - The first stereospecific, enantiospecific total synthesis of the ring-A oxygenated sarpagine indole alkaloids (+)-N(a)-methylsarpagine (8), (+)-majvinine (14), and (+)-10-methoxyaffinisine (49), as well as the first total synthesis of the Alstonia bisindole alkaloid macralstonidine (9), has been accomplished. This approach employed the Schollkopf chiral auxiliary for the stereospecific construction of the desired d-(+)-tryptophan unit required for the asymmetric Pictet-Spengler reaction. In addition, the strategy was doubly convergent for the enolate-mediated Pd(0) coupling process and the asymmetric Pictet-Spengler reaction can be employed to synthesize both macroline (2) and N(a) methylsarpagine (8), the coupling of which provides macralstonidine (9). This approach to ring-A substituted alkoxyindole alkaloids should find wide application for the synthesis of other alkaloids for it is stereospecific and either enantiomer can be prepared with ease. PMID- 12895063 TI - A prototype calix[4]arene-based receptor for carbohydrate recognition containing peptide and phosphate binding groups. AB - A novel class of macrobicyclic receptors for carbohydrate recognition based on upper rim, peptide-bridged calix[4]arenes has been designed and synthesized. Receptor 12, in which a charged phosphate group cooperates with peptide hydrogen bonding donor and acceptor groups in the binding process, is the most efficient and selective in the complexation of simple carbohydrate derivatives. The selectivity observed is toward beta-glucoside 13a, which is better bound (DeltaG degrees = 19.6 kJ mol(-)(1)) compared to the corresponding alpha anomer 13b (DeltaG degrees = 17.0 kJ mol(-)(1)) and to the beta-galactoside 13c (DeltaG degrees = 17.7 kJ mol(-)(1)) in CDCl(3). A substantial drop in the stability constant is observed by esterification of the phosphate group in the host 12 or by alkylation of the OH groups in the 2 and 3 positions in the beta-glucoside and beta-galactoside derivatives. On the basis of a careful analysis of the (1)H NMR data available, a binding mode of the beta-octylglucoside 13a to receptor 12 is proposed. PMID- 12895064 TI - Bent structure and dynamic stereochemistry of chiral acridinium cations. AB - Chiral acridinium cations, easily prepared by reaction of enantiopure primary amines and tris(2,6-dimethoxy)trityl cation, display restricted rotations around the C(sp(3))-N(sp(2)) bond. A bending of the aromatic backbone and "out-of-plane" displacements of C(9) and N(10) atoms are enforced if bulky amines are introduced. PMID- 12895065 TI - Total synthesis of the antiallergic naphtho-alpha-pyrone tetraglucoside, cassiaside C(2), isolated from cassia seeds. AB - Toralactone 9-O-beta-d-glucopyranosyl-(1-->6)-beta-d-glucopyranosyl-(1-->3)-beta d-glucopyranosyl-(1-->6)-beta-d-glucopyranoside (1, cassiaside C(2)), isolated from Cassia obtusifolia L. and showing strong antiallergic activity, was concisely synthesized employing glycosyl trifluoroacetimidates as glycosylation agents. The unique naphtho-alpha-pyrone structure of toralactone (5) was constructed by condensation of orsellinate 8 with pyrone 9 in the presence of LDA as developed by Staunton and co-workers. The naphthol of toralactone showed minimal reactivity as an acceptor and was screened with various glycosyl donors. It is finally concluded that sacrifice of an excess amount of the trifluoroacetimidate or trichloroacetimidate donors (6f/6g, 6.0 equiv) in the presence of a catalytic amount of TMSOTf (0.05 and 0.3 equiv, respectively) afforded excellent yields of the coupling product, which was otherwise only a minor product under a variety of conditions examined. PMID- 12895066 TI - An efficient approach to dihydrofurocoumarins via palladium-catalyzed annulation of 1,3-dienes by o-iodoacetoxycoumarins. AB - The palladium-catalyzed annulation of 1,3-dienes by o-iodoacetoxycoumarins provides an efficient method for the synthesis of biologically interesting dihydrofurocoumarins. The presence of the acetyl group on the phenolic oxygen and the use of silver carbonate as a base are crucial for this process. This reaction is very general and regio- and stereoselective, and a wide variety of terminal, cyclic, and internal 1,3-dienes can be utilized. Derivatization of the annulation products provides an efficient approach to numerous analogues of natural products. PMID- 12895067 TI - Investigation on the regioselectivities of intramolecular oxidation of unactivated C-H bonds by dioxiranes generated in situ. AB - We found that dioxiranes generated in situ from ketones 1-6 and Oxone underwent intramolecular oxidation of unactivated C-H bonds at delta sites of ketones to yield tetrahydropyrans. From the trans/cis ratio of oxidation products 1a and 2a as well as the retention of the configuration at the delta site of ketone 5, we proposed that the oxidation reaction proceeds through a concerted pathway under a spiro transition state. The intramolecular oxidation of ketone 6 showed the preference for a tertiary delta C-H bond over a secondary one. This intramolecular oxidation method can be extended to the oxidation of the tertiary gamma' C-H bond of ketones 9 and 10. For ketone 11 with two delta C-H bonds and one gamma' C-H bond linked respectively by a sp(3) hydrocarbon tether and a sp(2) ester tether, the oxidation took place exclusively at the delta C-H bonds. Finally, by introducing proper tethers, regioselective hydroxylation of steroid ketones 12-14 have been achieved at the C-17, C-16, C-3, and C-5 positions. PMID- 12895068 TI - A new synthetic route to enantiomerically pure axially chiral 2,2'-bipyridine N,N'-dioxides. Highly efficient catalysts for asymmetric allylation of aldehydes with allyl(trichloro)silanes. AB - New axially chiral 2,2'-bipyridine N,N'-dioxides 1 were obtained in an enantiomerically pure form by way of cyclic diesters 6 or 7 which were formed by the esterification of diols 2 with (R)-2,2'-bis(chlorocarbonyl)-1,1' binaphthalene (5). Epimerization of the kinetic products at the ester formation (R(nap),S(pyr))-6 to the thermodynamically stable isomers (R(nap),R(pyr))-7 was observed in refluxing toluene or in the presence of trifluoroacetic acid. One of the N,N'-dioxides 1a which is substituted with phenyl groups at the 6 and 6' positions was found to be highly catalytically active and enantioselective for the asymmetric allylation of aldehydes with allyl(trichloro)silane giving homoallyl alcohols. PMID- 12895069 TI - 1,3-dipolar cycloaddition of 2-dialkylaminothioisomunchnones with aliphatic aldehydes: synthesis of beta-lactams and thiiranes, structure elucidation, and rationale for chemoselective fragmentation of cycloadducts. AB - A series of highly funtionalized beta-lactams and thiiranes can be generated on treatment of 1,3-thiazolium-4-olates (thioisomunchnones) with aliphatic aldehydes. Although in some cases a variety of products have been obtained, the present paper now provides a mechanistic rationale to explain the product distribution based on stereoelectronic effects. Thus, ring fragmentation of the initial [3+2] cycloadduct is essentially dictated by the electronic character of the aryl substituent on the nitrogen atom of the parent thioisomunchnone. However, further evolution of such cycloadducts into beta-lactams or thiiranes is governed by steric effects to a large extent. Evidence for such interactions has been obtained by computing PM3-optimized diastereomeric transition structures in the reaction of a thioisomunchnone with a chiral aliphatic aldehyde. PMID- 12895070 TI - Evidence of an electron-transfer mechanism in the peroxynitrite-mediated oxidation of 4-alkylphenols and tyrosine. AB - The mechanism of interaction of the peroxynitrite with some 4-alkylphenols and tyrosine was mainly studied by means of ESR spectroscopy and product analysis. The radical intermediates, detected as spin adducts to the 5-diethoxyphosphoryl-5 methyl-1-pyrroline N-oxide (DEPMPO), were identified as carbon-centered radicals to the benzene ring. The reaction seems to proceed via an electron transfer process (ET), most likely mediated by a NOx derivative, leading to the intermediacy of a phenoxyl-type radical as proved by the detection of the corresponding Pummerer-type ketone. No evidence of the formation of hydroxyl radicals, due to the homolytic cleavage of the peroxynitrite at physiological pH was obtained, even though DEPMPO hydroxyl spin adducts were detected: the latter most likely arises from the direct attack of the spin trap by the oxidant species. The possible involvement of HCO(3)(-)/CO(2), i.e., the formation of the nitrosoperoxycarbonate, ONOOCO(2)(*)(-), was also investigated. PMID- 12895071 TI - Cross-coupling reactions of (1-fluorovinyl)methydiphenylsilane(1) with aryl halides and aryl triflates. AB - The cesium fluoride (CsF)-assisted cross-coupling reaction of (1 fluorovinyl)methyldiphenylsilane (1) with aryl halides and aryl triflates was examined. The reaction with aryl iodides smoothly proceeded to afford the corresponding (1-fluorovinyl)arenes in the presence of a catalytic amount of CuI and Pd(PPh(3))(4) in aprotic polar solvents such as DMF, DMI, DMA, and NMP in good yields. A variety of functional groups (nitro, ester, ketone, and ether) on the aromatic rings can be tolerated under these mild conditions. Aryl iodides are superior to aryl bromides as the coupling reaction partner. The cross-coupling reaction of 1 with aryl triflates instead of aryl halides was also accomplished in the presence of tetrabutylammonium iodide (n-Bu(4)NI) as the additive under similar conditions. PMID- 12895072 TI - Preparation and cycloaddition reactions of enantiopure 2-(N-acylamino)-1,3-dienes for the synthesis of octahydroquinoline derivatives. AB - Stille, Suzuki-Miyaura, and Sonogashira cross-coupling reactions were carried out with a glutarimide-derived vinyl phosphate, bearing a chiral auxiliary on the N atom, to prepare enantiopure 2-(N-acylamino)-1,3-dienes as partners in Diels Alder reactions. The cycloadditions were performed with various dienophiles under thermal conditions, with or without Lewis acids. With maleimides, the preferential formation of endo cycloadducts was observed, whereas with acrylamides the exo approach prevailed. Furthermore, in the latter case, 6 substitued octahydroquinolinones were obtained in accordance with the predicted regioselectivity. Since diastereopure compounds were in all cases obtained either by chromatography or by crystallization, and because of the easy access to a variety of boronic acids, to be used in the coupling step, this methodology is useful for the short synthesis of differently substituted, enantiopure octahydroquinolinones amenable to further transformation into decahydroquinolines possessing interesting biological activities. PMID- 12895073 TI - A theoretical study on the mechanism of the cyclopolymerization of diallyl monomers. AB - The cyclization and intermolecular propagation steps of the cyclopolymerization mechanism are studied with density functional theory. In addition to standard cyclization and intermolecular propagation reactions of cyclopolymerization, competing reactions that lead to chain transfer and termination are also discussed. The mechanistic study of the cyclopolymerization reaction of two representative monomers, N,N-diallylamine (1) and N,N-dimethyl-N,N-diallylamonium (2), was carried out with B3LYP/6-31G computations. Monomer 1 has almost the same activation barriers for homopolymerization and cyclization. In monomer 2, cyclization is much more facile than homopolymerization, leading to the higher cyclopolymerization efficiency. In the case of 2, methyl substituents on nitrogen inhibit hydrogen abstraction, whereas in 1, hydrogen abstraction reactions from the neutral monomer yield stabilized products leading to chain transfer. Calculations show that facile competing reactions of monomer 1 lower the polymerization efficiency. Monomer 2 displays a stronger preference for cyclization relative to other processes. PMID- 12895074 TI - Solvation effects on alternative nucleophilic substitution reaction paths for chloride/allyl chloride and gamma-methylated congeners. AB - An adiabatic connection method, mPW1PW91(0.581)/6-31G(d), was employed in conjunction with the continuum solvation model SM5.42 to study the effects of solvation on the S(N)1, S(N)2, and syn and anti S(N)2' nucleophilic substitution reactions of chloride anion with allyl chloride and its gamma-methylated analogues Z- and E-crotyl chloride and isoprenyl chloride. The impact of equilibrium solvation on the potential energy surfaces of these systems is large and leads to significant changes in both the geometries and the relative energetics of different reaction pathways for different species. The predicted effects of increased solvent dielectric constant are consistent with available experimental data and provide semiquantitative insights into the relative influence of different solvents on particular properties. PMID- 12895075 TI - Ketone enolization with lithium dialkylamides: the effects of structure, solvation, and mixed aggregates with excess butyllithium. AB - The effects of lithium dialkylamide structure, mixed aggregate formation, and solvation on the stereoselectivity of ketone enolization were examined. Of the lithium dialkylamides examined, lithium tetramethylpiperidide (LiTMP) in THF resulted in the best enolization selectivity. The stereoselectivity was further improved in the presence of a LiTMP-butyllithium mixed aggregate. The use of less polar solvents reduced the enolization stereoselectivity. Ab initio calculations predict LDA and LiTMP to form mixed cyclic dimers in ethereal solvents. The calculations also predict LiTMP-alkyllithium mixed aggregates to competitively inhibit the formation of less stereoselective LiTMP-lithium enolate mixed aggregates. PMID- 12895076 TI - Application of a new tandem isomerization-aldolization reaction of allylic alcohols to the synthesis of three diastereoisomers of (2R)-1,2-O-isopropylidene 4-methylpentane-1,2,3,5-tetraol. AB - The tandem isomerization-aldolization reaction of (2R)-1,2-O-isopropylidene-4 penten-1,2,3-triol 3 and formaldehyde gives a mixture of two aldol products 2a and 2b. The stereoselective reduction of each compound by l-Selectride affords two diastereoisomers of (2R)-1,2-O-Isopropylidene-4-methylpentane-1,2,3,5-tetraol while a third diastereoisomer is obtained by stereoselective reduction with Me(4)NHB(OAc)(3). PMID- 12895077 TI - Synthesis of GDP-5-thiosugars and their use as glycosyl donor substrates for glycosyltransferases. AB - Two thiopyranoside analogues of GDP-sugars, GDP-5-thio-d-mannose (14) and GDP-5 thio-l-fucose (15), were synthesized. The syntheses included the phosphorylations of tetra-O-acetyl-5-thio-d-mannosyl bromide (4) and tri-O-benzoyl-l-fucosyl bromide (6) with silver dibenzyl phosphate, deprotection of the phosphate groups, and condensation of the deprotected phosphates with GMP-imidazolidate (13) in the presence of MgCl(2). These GDP-sugar analogues were found to be donor substrates for alpha(1,2)mannosyltransferase and alpha(1,3)fucosyltransferase, affording a 5 thiomannose-containing disaccharide (18) and a 5-thiofucose-containing trisaccharide (21), respectively. The conformation of the disaccharide analogue 18 was similar to that of its native counterpart by ROESY. These findings for GDP 5-thiosugars together with previous demonstrations of enzymatic transfer from UDP 5-thiosugars will allow the production of panels of oligosaccharide analogues with hydrolase-resistant properties. PMID- 12895078 TI - Ring opening of nonactivated 2-(1-aminoalkyl) aziridines: unusual regio- and stereoselective C-2 and C-3 cleavage. AB - We have studied the ring opening of nonactivated amino aziridines 1 by water under acidic conditions. Depending on the acid used, amino aziridines are cleaved at C-3 or C-2 with high regioselectivity, and total stereoselectivity, affording chiral 2,3-diaminoalkan-1-ols 3 or 1,3-diaminoalkan-2-ols 4 in high yield. PMID- 12895079 TI - Selectivity of purine alkylation by a quinone methide. Kinetic or thermodynamic control? AB - The alkylation reaction of 9-methyladenine and 9-methylguanine (as prototype substrates of deoxy-adenosine and -guanosine), by the parent o-quinone methide (o QM), has been investigated in the gas phase and in aqueous solution, using density functional theory at the B3LYP/6-311+G(d,p) level. The effect of the medium on the reactivity, and on the stability of the resulting adducts, has been investigated by using the C-PCM solvation model to assess which adduct arises from the kinetically favorable path, or from an equilibrating process. The calculations indicate that the most nucleophilic site of the methyl-substituted nucleobases in the gas phase is the guanine oxygen atom (O(6)) (DeltaG()(gas) = 5.6 kcal mol(-)(1)), followed by the adenine N1 (DeltaG)(gas) = 10.3 kcal mol( )(1)), while other centers exhibit a substantially lower nucleophilicity. The bulk effect of water as a solvent is the dramatic reduction of the nucleophilicity of both 9-methyladenine N1 (DeltaG)(solv) = 14.5 kcal mol(-)(1)) and 9-methylguanine O(6) (DeltaG)(solv) = 17.0 kcal mol(-)(1)). As a result there is a reversal of the nucleophilicity order of the purine bases. While O(6) and N7 nucleophilic centers of 9-methylguanine compete almost on the same footing, the reactivity gap between N1 and N7 of 9-methyladenine in solution is highly reduced. Regarding product stability, calculations predict that only two of the adducts of o-QM with 9-methyladenine, those at NH(2) and N1 positions, are lower in energy than reactants, both in the gas phase and in water. However, the adduct at N1 can easily dissociate in water. The adducts arising from the covalent modification of 9-methylguanine are largely more stable than reactants in the gas phase, but their stability is markedly reduced in water. In particular, the oxygen alkylation adduct becomes slightly unstable in water (DeltaG(solv) = +1.4 kcal mol(-)(1)), and the N7 alkylation product remains only moderately more stable than free reactants (DeltaG(solv) = -2.8 kcal mol(-)(1)). Our data show that site alkylations at the adenine N1 and the guanine O(6) and N7 in water are the result of kinetically controlled processes and that the selective modification of the exo-amino groups of guanine N2 and adenine N6 are generated by thermodynamic equilibrations. The ability of o-QM to form several metastable adducts with purine nucleobases (at guanine N7 and O(2), and adenine N1) in water suggests that the above adducts may act as o-QM carriers. PMID- 12895080 TI - Novel oxidative alpha-tosyloxylation of alcohols with iodosylbenzene and p toluenesulfonic acid and its synthetic use for direct preparation of heteroaromatics. AB - alpha-Tosyloxyketones and alpha-tosyloxyaldehydes were directly prepared from alcohols by treatment with iodosylbenzene and p-toluenesulfonic acid monohydrate in good yields. This method can be used for the direct preparation of thiazoles, imidazoles, and imidazo[1,2-a]pyridines from alcohols in good to moderate yields by the successive treatment with iodosylbenzene and p-toluenesulfonic acid monohydrate, followed by thioamides, benzamidine, and 2-aminopyridine, respectively. PMID- 12895081 TI - Synthesis of 3-substituted quinolines via transition-metal-catalyzed reductive cyclization of o-nitro Baylis-Hillman acetates. AB - Reductive cyclization of o-nitro-substituted Baylis-Hillman acetates by carbon monoxide, catalyzed by [CpFe(CO)(2)](2), gives moderate to good yields of 3 substituted quinolines. PMID- 12895082 TI - Rapid microwave promoted Sonogashira coupling reactions on solid phase. AB - A microwave-enhanced, rapid, and efficient solid-phase version of the Sonogashira reaction is presented. It has been applied to the coupling of aryl iodides and bromides with various acetylene derivatives giving excellent yields in 15-25 min. The scopes of homogeneous, solventless, and solid-phase conditions for Sonogashira coupling of aryl halides are compared. PMID- 12895083 TI - Direct synthesis of 1,1'-[1,4-phenylenebis(methylene)]-bis- 1,4,8,11 tetraazacyclotetradecane octahydrochloride (AMD 3100) without the use of protecting groups. AB - A four-step synthesis of the title compound starting from methyl acrylate, ethylenediamine, and dimethyl malonate is reported. The synthesis can be run on a multigram scale and is operationally simple. The use of protecting groups is avoided by utilizing the trioxocyclam as the key coupling intermediate. PMID- 12895084 TI - First enantioselective total synthesis of (8S,12R,15S)-prostaglandin J(2). AB - Enantioselective synthesis of natural PGJ(2) has been accomplished for the first time starting from the commercially available enantiopure aldehyde 7 in 10% overall yield. The key reaction was a novel prostaglandin class interconversion, i.e., an allylic 1,3-transposition across alcohol 9 derived from compound 14 in 73% overall yield. In principle, the unnatural enantiomer of PGJ(2) could be obtained starting from the commercially available enantiopure monobenzoate 7a following our strategy. PMID- 12895085 TI - Preparation of protected syn-alpha,beta-dialkyl beta-amino acids that contain polar side chain functionality. AB - We report the synthesis of syn-alpha,beta-dialkyl beta-amino acid derivatives suitably protected for solid-phase synthesis that give rise to residues containing positively charged lysine-like side chains. These amino acids, as well as syn-alpha,beta-dialkyl beta-amino acids that contain diverse hydrophobic side chains, are prepared in good de and ee. The key step in this route involves Davies's protocol for the conjugate addition of a chiral lithium amide to alpha,beta-unsaturated tert-butyl esters (Davies, S. G.; Ichihara, O.; Walters, I. A. S. J. Chem. Soc., Perkin Trans. 1 1994, 9, 1141). syn-alpha,beta-Dialkyl beta-amino acids are interesting building blocks because of their sheet-forming propensity and because of their presence in bioactive compounds. PMID- 12895086 TI - Stereoselective Barbier-type allylation reaction of trifluoromethyl aldimines. AB - Trifluoromethyl aldimines could react, under Barbier conditions in the presence of activated zinc, in DMF at room temperature or in THF at reflux, with various allyl bromides to provide the corresponding homoallylamines. Secondary homoallyl trifluoromethylamines were stereoselectively obtained from the optically active aldimine 12 with an excellent diastereoisomeric excess (98%). PMID- 12895087 TI - Early amidation approach to 3-[(4-amido)pyrrol-2-yl]-2-indolinones. AB - A new synthesis of 3-[(4-amido)pyrrol-2-yl]-2-indolinones has been developed, where the amide side chain was installed prior to pyrrole formation. This strategy precludes the need to use any coupling reagents to install the amide side chain. This process includes a zinc-free alternative to the Knorr pyrrole synthesis. PMID- 12895088 TI - Stereocontrol in the EtAlCl(2)-induced cyclization of chiral gamma,delta unsaturated methyl ketones to form cyclopentanones. AB - EtAlCl(2)-induced cyclization of chiral gamma,delta-unsaturated ketones 11c and 17b takes place mainly from the expected face. The selectivity is modest for 11c (60:40) in which the large substituent is a primary alkyl group and the medium substituent is a methyl group and excellent for 17b (93:7) in which the large substituent is a cyclohexyl group and the medium substituent is a methyl group. The cyclization of 17a is anomalous, suggesting that the phenyl group has more than a simple steric effect. PMID- 12895089 TI - Expeditious copper-catalyzed conjugate 1,4-addition of bromo[2-(1,3-dioxolan-2 yl)ethyl]magnesium to alpha,beta-cycloalkenones and subsequent transformations. AB - Expeditious CuI-catalyzed conjugate 1,4-addition of bromo[2-(1,3-dioxolan-2 yl)ethyl]magnesium to the five-, six-, seven-, and eight-membered alpha,beta cycloalkenones is described. The reaction times are decreased dramatically compared to CuBr-(CH(3))(2)S catalysis. The resulting ketoacetals were subsequently cyclized to bicyclic beta-hydroxy ketones and alpha,beta-enones, followed by further transformations. PMID- 12895090 TI - Nucleation of beta-hairpin structures with cis amide bonds in E-vinylogous proline-containing peptides. AB - Synthesis and conformational studies of peptides containing the E-vinylogous prolines 1 (VPro1) and 2 (VPro2), Boc-Ala-Val-VPro1-Xaa-Leu-OMe (3, Xaa = Gly; 4, Xaa = Phe), Boc-Ala-Val-VPro2-Xaa-Leu-OMe (5, Xaa = Gly; 6, Xaa = Phe), Boc-Leu Ile-Val-VPro1-Xaa-Leu-OMe (7, Xaa = Gly; 8, Xaa = Phe), and Boc-Leu-Ile-Val-VPro2 Xaa-Leu-OMe (9, Xaa = Gly; 10, Xaa = Phe), were carried out. It has been shown that both VPro1 and VPro2 lead to the formation of 12-membered intramolecularly hydrogen bonded structures very similar to type VI beta-turns with a cis Xaa-VPro amide bond in the major conformers in all the peptides 3-10, resulting in the nucleation of beta-hairpin type structures in these molecules in CDCl(3). PMID- 12895091 TI - A phosphine-catalyzed [3+2] cycloaddition strategy leading to the first total synthesis of (-)-hinesol. AB - In one step, the skeleton of cis-spirovetivanes was constructed with high stereoselectivity by the phosphine-catalyzed [3+2] cycloaddition reaction of tert butyl 2,3-butadienoate or 2-butynoate with 3-methyl-2-methylenecyclohexanone (5). This method was exemplified by the first highly efficient total synthesis of natural product (-)-hinesol, which is an active ingredient of cerebral circulation and metabolism improvers. PMID- 12895092 TI - The synthesis of 4-deazaformycin A. AB - The preparation of 4-deazaformycin A has been achieved. The synthesis features the condensation of a suitably substituted, lithiated 4-picoline with 2,3,5-tri-O benzyl-d-ribonolactone, dehydration of the resulting hemiacetal, and ionic hydrogenation, followed by manipulation of the protecting groups and subsequent ring closure with the formation of 7-amino-3-(beta-d-ribofuranosyl)pyrazolo[3,4 c]pyridine. PMID- 12895093 TI - Role of face-to-face and edge-to-face aromatic interactions in the inclusion complexation of cyclobis(paraquat-p-phenylene): a theoretical study. AB - A B3LYP/6-31G(d,p) and MP2/6-31G(d,p)//B3LYP/6-31G(d,p) computational study of the gas-phase complexation of cyclobis(paraquat-p-phenylene) (1(4+)) with four typical aromatic guests, namely, 1,4-dimethoxybenzene (2), 1,5 dimethoxynaphthalene (3), benzidine (4), and tetrathiafulvalene (5), has been carried out. The structure of the host has been successively split into two responsible substructures, respectively, for the face-to-face and edge-to-face interactions with the guests. The sum of the two interactions calculated at the B3LYP/6-31G(d,p) and MP2/6-31G(d,p)//B3LYP/6-31G(d,p) levels for each guest proved to be in good agreement with the overall binding energy of the host calculated at the corresponding level of theory. The results show that the binding of the complexes is primarily due to London dispersion interactions which require wave function-based correlation methods for an adequate description. Face to-face interactions are about 1 order of magnitude more important than edge-to face interactions in determining the overall binding energy. While edge-to-face interactions essentially depend on London dispersion forces, face-to-face interactions depend about one-half on electrostatic and frontier orbital contributions (the latter being more important) and the other half on London dispersion forces. PMID- 12895095 TI - Mononuclear N3S(thioether)-ligated copper(II) methoxide complexes: synthesis, characterization, and hydrolytic reactivity. AB - Mononuclear copper(II) methoxide complexes supported by N(3)S(thioether) chelate ligands having two internal hydrogen bond donors have been prepared, comprehensively characterized, and evaluated for hydrolytic reactivity. PMID- 12895096 TI - Ferromagnetic ordering and metamagnetism in malonate bridged 3D diamond-like and honeycomb-like networks: [Cu(mal)(DMF)]n and [[Cu(mal)(0.5pyz)].H2O]n (mal = Malonate Dianion, DMF = N,N-dimethylformamide, pyz = Pyrazine). AB - Two three-dimensional (3D) malonate bridged networks, [Cu(mal)(DMF)](n) (1) and [[Cu(mal)(0.5pyz)].H(2)O](n) (2), have been synthesized in H(2)O-DMF solution. Compound 1 exhibits ferromagnetic ordering below 2.6 K (T(c)), and 2 displays a metamagnetic behavior below 3.2 K (T(N)). PMID- 12895097 TI - Self-assembly of octarhenium-based neutral luminescent rectangular prisms. AB - Self-assembly of rhenium-based nanoscale rectangular prismatic boxes has been achieved in quantitative conversion. The fac-rhenium corner provided three mutually perpendicular coordination sites and served as a good candidate for the construction of 3-D boxes. These are the first Re-based, neutral, luminescent prisms of M(8)L(2)L'(8) type that has been characterized crystallographically. Their luminescent properties and molecular recognition capabilities make these molecular prisms interesting supramolecules. PMID- 12895098 TI - Utilization of CS2 as a source of C1 chemistry for the generation of methyldithioformate. AB - Insertion of CS(2) into the Ru-H bond of cis-[(dppe)(2)Ru(H)(2)] takes place to afford the hydride dithioformate complex trans-[(dppe)(2)Ru(H)(SC(S)H)]. The hydride dithioformate complex reacts under very mild conditions with MeX (X = OTf, I) to give the hydride methyldithioformate derivative trans [(dppe)(2)Ru(H)(SC(SMe)H)][X]. Three different pathways have been found to cleave off the ester moiety from the metal complex. A method to recover the ruthenium starting material upon elimination of the methyldithioformate is presented. This is a novel case of C(1) chemistry using carbon disulfide. PMID- 12895099 TI - Structural studies of transition metal complexes with 4,5-bis(2 pyridylmethylsulfanyl)-4',5'-ethylenedithiotetrathiafulvalene: probing their potential for the construction of multifunctional molecular assemblies. AB - Three divalent transition metal complexes of 4,5-bis(2-pyridylmethylsulfanyl) 4',5'-ethylenedithiotetrathiafulvalene have been prepared and crystallographically characterized. The isostructural Co(II) and the Ni(II) complexes show octahedral geometries around the metal ions with the coordination sites occupied by the pyridyl nitrogen atoms and the thioether sulfur atoms of the ligand and cis coordination of the halide ions. Cyclic voltammetry reveals that the complexation leads to a small anodic shift in the first oxidation potential of the TTF system. PMID- 12895100 TI - The first example of a monomeric alumatrane. AB - The novel five-coordinate aluminum adduct Me(2)HN. AlL (2) [wherein L = tris(2 oxy-3,5-dimethylbenzyl)amine] containing three six-membered rings has been characterized by spectroscopic and by X-ray means. This adduct of an alumatrane is the first structurally characterized monomeric alumatrane derivative, and unlike its parent alumatrane [Al(OCH(2)CH(2))(3)N](x), 2 is monomeric in the gas, solution, and solid states. The X-ray molecular structure of 2 reveals a tricyclic cage moiety of C(3) symmetry. The aluminum geometry is a slightly distorted trigonal bipyramid in which, quite unexpectedly, the metal atom is located somewhat below the plane formed by three equatorial oxygens and its Al N(tertiary) bond is shorter than that in Me(3)N.AlH(3).NMe(3) (Heitsch, C. W.; Nordman, C. E.; Parry, R. W. Inorg. Chem. 1963, 2, 508). PMID- 12895101 TI - Effect of electron availability on selectivity of O2 reduction by synthetic monometallic Fe porphyrins. AB - Herein we report that biomimetic analogues of cytochrome c oxidase (CcO) couple reduction of O(2) to oxidation of a single-electron carrier, Ru(NH(3))(6)(2+), under steady-state catalytic turnover. Higher Ru(II) concentrations favor the 4 electron vs 2-electron O(2) reduction pathway. Our data indicate that the capacity of electrode-adsorbed Fe-only porphyrins to catalyze reduction of O(2) to H(2)O is due to high availability of electrons and is eliminated under the biologically relevant slow electron delivery. PMID- 12895102 TI - "Chiral-at-Metal" octahedral ruthenium(II) complexes with achiral ligands: a new type of enantioselective catalyst. AB - cis-[Ru(dmp)(2)(CH(3)CN)(2)][PF(6)](2) (dmp = 2,9-dimethyl-1,10-phenanthroline), complex 1[PF(6)](2), exists in two enantiomeric forms, Delta and Lambda. During treatment with the chiral anion tris[tetrachlorobenzene-1,2 bis(olato)]phosphate(V), also named Trisphat, in dichloromethane it has been possible to selectively precipitate each enantiomer, associated with Trisphat in the form of the heterochiral pair. This enantiomerically pure compound has been characterized in solution by UV-visible, CD, ESI-MS, and NMR spectroscopy and by X-ray crystallography in the solid state. Trisphat was also used as an NMR chiral shift reagent to determine the enantiomeric excess of the complex preparations. The "chiral-at-metal" ruthenium complex has been evaluated as a catalyst for the oxidation of sulfides to sulfoxides by hydrogen peroxide. The reactions displayed a low but significant level of enantioselectivity (18% ee in the case of 4 bromophenyl methyl sulfide). Our results thus provide the first demonstration that the chiral information carried by a stereogenic metal center can be catalytically transferred to molecules during stereoselective oxidation. PMID- 12895103 TI - Binuclear manganese compounds of potential biological significance. Part 2. Mechanistic study of hydrogen peroxide disproportionation by dimanganese complexes: the two oxygen atoms of the peroxide end up in a dioxo intermediate. AB - The dimanganese(II,II) complexes 1a [Mn(2)(L)(OAc)(2)(CH(3)OH)](ClO(4)) and 1b [Mn(2)(L)(OBz)(2)(H(2)O)](ClO(4)), where HL is the unsymmetrical phenol ligand 2 (bis-(2-pyridylmethyl)aminomethyl)-6-((2-pyridylmethyl)(benzyl)aminomethyl)-4 methylphenol, react with hydrogen peroxide in acetonitrile solution. The disproportionation reaction was monitored by electrospray ionization mass spectrometry (ESI-MS) and EPR and UV-visible spectroscopies. Extensive EPR studies have shown that a species (2) exhibiting a 16-line spectrum at g approximately 2 persists during catalysis. ESI-MS experiments conducted similarly during catalysis associate 2a with a peak at 729 (791 for 2b) corresponding to the formula [Mn(III)Mn(IV)(L)(O)(2)(OAc)](+) ([Mn(III)Mn(IV)(L)(O)(2)(OBz)](+) for 2b). At the end of the reaction, it is partly replaced by a species (3) possessing a broad unfeatured signal at g approximately 2. ESI-MS associates 3a with a peak at 713 (775 for 3b) corresponding to the formula [Mn(II)Mn(III)(L)(O)(OAc)](+) ([Mn(II)Mn(III)(L)(O)(OBz)](+) for 3b). In the presence of H(2)(18)O, these two peaks move to 733 and to 715 indicating the presence of two and one oxo ligands, respectively. When H(2)(18)O(2) is used, 2a and 3a are labeled showing that the oxo ligands come from H(2)O(2). Interestingly, when an equimolar mixture of H(2)O(2) and H(2)(18)O(2) is used, only unlabeled and doubly labeled 2a/b are formed, showing that its two oxo ligands come from the same H(2)O(2) molecule. All these experiments lead to attribute the formula [Mn(III)Mn(IV)(L)(O)(2)(OAc)](+) to 2a and to 3a the formula [Mn(II)Mn(III)(L)(O)(OAc)](+). Freeze-quench/EPR experiments revealed that 2a appears at 500 ms and that another species with a 6-line spectrum is formed transiently at ca. 100 ms. 2a was prepared by reaction of 1a with tert butyl hydroperoxide as shown by EPR and UV-visible spectroscopies and ESI-MS experiments. Its structure was studied by X-ray absorption experiments which revealed the presence of two or three O atoms at 1.87 A and three or two N/O atoms at 2.14 A. In addition one N atom was found at a longer distance (2.3 A) and one Mn at 2.63 A. 2a can be one-electron oxidized at E(1/2) = 0.91 V(NHE) (DeltaE(1/2) = 0.08 V) leading to its Mn(IV)Mn(IV) analogue. The formation of 2a from 1a was monitored by UV-visible and X-ray absorption spectroscopies. Both concur to show that an intermediate Mn(II)Mn(III) species, resembling 4a [Mn(2)(L)(OAc)(2)(H(2)O)](ClO(4))(2), the one-electron-oxidized form of 1a, is formed initially and transforms into 2a. The structures of the active intermediates 2 and 3 are discussed in light of their spectroscopic properties, and potential mechanisms are considered and discussed in the context of the biological reaction. PMID- 12895104 TI - Formation of two diverse classes of poly(amino-alkoxide) chelates and their mononuclear and polynuclear lanthanide(III) complexes. AB - Factors that influence aggregation of lanthanide(III) (Ln(III)) ions to form polynuclear complexes were studied utilizing 1-aziridineethanol as a versatile source of macrocyclic and acyclic chelates. The facile ring-opening cyclo oligomerization of 1-aziridineethanol leads to the formation of a series of polyaza cyclic oligomers (series A). In the presence of ethylenediamine, a competing N-alkylation reaction occurs to produce a new class of acyclic ligands (series B). The cyclo-oligomerization of four 1-aziridineethanol units is the most favorable process, leading to the formation of the 12-membered cyclen-type macrocycle, H(4)L(1) (1,4,7,10-tetrakis(2-hydroxyethyl)-1,4,7,10-tetraaza cyclododecane). Ring-opening cyclo-oligomerization of 1-aziridineethanol in the presence of Ln(III) ions produces self-assembled mononuclear, tetranuclear, and pentanuclear compounds of H(4)L(1). In the presence of ethylenediamine, oligomerization of 1-aziridineethanol results in a dinuclear complex of an acyclic poly(amino-alkoxide) H(2)L(2). The coordinative unsaturation of (i) the alkoxy sites of [H(x)L(1)](x)(-)(4) (where x < 4) and (ii) Ln(III) ions in coordination numbers less than nine are critical factors in the formation of the polynuclear Ln(III) complexes. The identities of mononuclear, dinuclear, tetranuclear, and pentanuclear complexes herein discussed were established by X ray crystallography. PMID- 12895105 TI - An electron paramagnetic resonance study of copper(II)-beta-substituted beta amino acid systems by the two-dimensional simulation method: first evidence of primarily steric effects of substituents on equilibria of metal complexes. AB - We have studied the complex equilibria of copper(II) with a series of beta substituted beta-amino acids (R: H, Me, Et, iBu, iPr, cHex, 1-EtPr, and tBu) in aqueous solution by pH potentiometry and electron paramagnetic resonace (EPR) spectroscopy in the range pH = 2-8 at various metal and ligand concentrations. The basicities of the corresponding donor groups differed only slightly in the series of ligands. A purely mathematical method, the matrix rank analysis carried out on the EPR spectrum package recorded in the presence of copper(II), indicated the formation of 6 independent paramagnetic species. Accordingly, Cu(2+) (aqua complex) and the complexes [CuLH](2+), [CuL](+), [CuL(2)H(2)](2+), [CuL(2)H](+), and [CuL(2)] were considered in the subsequent analysis of series of spectra, and also two isomers of [CuL(2)] were identified. The formation constants and the EPR parameters, e.g. the isotropic g-factors and the copper and nitrogen hyperfine couplings for the above species, were determined in the same optimization procedure by the simultaneous evaluation of spectra. The ligands "LH" are suggested to bind in equatorial positions through their carboxylate groups, while the amino acids in the L protonation state are likely to occupy two equatorial sites via the amino and carboxylate groups. For the isomers of [CuL(2)], the donors of the same kind are in the cis or trans position. As far as we know, this is the first reported case in which a strong correlation has been found between the steric effects of substituents characterized by Meyer's steric parameter V(a) and the protonation constants of metal complexes. The observed trend for the preference for nonprotonated complexes [CuL](+) and [CuL(2)] to increase with the steric demand of the substituent was explained by the increasing shielding effect of the substituent hindering protonation of the nonprotonated complex. PMID- 12895106 TI - A compelling experimental test of the hypothesis that enzymes have evolved to enhance quantum mechanical tunneling in hydrogen transfer reactions: the beta neopentylcobalamin system combined with prior adocobalamin data. AB - An intriguing but controversial hypothesis has appeared that "The optimization of enzyme catalysis may entail the evolutionary implementation of chemical strategies that increase the probability of tunneling and thereby accelerate the reaction rate" (Kohen, A.; Klinman, J. P. Acc. Chem. Res. 1998, 31, 397). Restated, enzymes may have evolved to enhance quantum mechanical tunneling by coupling to protein low nu modes that squeeze the reacting centers together in, for example, their H(*) atom abstraction reactions. Such a putative "protein squeezing" mechanism would enhance hydrogen quantum mechanical tunneling by reducing the barrier width. An alternative hypothesis is that enzymes do not enhance tunneling, but simply exploit the same amount of tunneling present in their enzyme-free solution reactions, if those reactions occur. A third, conceivable hypothesis is that enzymes might even inadvertently decrease the amount of tunneling as an undesired result of increasing the barrier width while reducing the barrier height. Testing these hypotheses experimentally requires the extremely rare event of being able to measure the amount of tunneling both in the enzyme system and in a very similar if not identical reaction in enzyme-free solution. This has been accomplished experimentally in only one prior case, our recent study of AdoCbl (coenzyme B(12)) and 8-Meo-AdoCbl undergoing enzyme-like H(*) abstraction reactions (Doll, K. M.; Bender, B. R.; Finke, R. G. to J. Am. Chem. Soc. 2003, in press). The data there reveal no change in the level of tunneling within or outside of the enzyme in comparison to the best literature data for an AdoCbl-dependent enzyme, methylmalonyl-CoA mutase. However, that first system suffers from two limitations: the measurement of the KIE (kinetic isotope effect) data in a nonenzymic 80-110 degrees C temperature range; and lower precision data than desired due to the HPLC-MS method required for one of the KIE analyses. These limitations have now been overcome by the synthesis, then thermolysis and KIE study vs temperature of the H(*) abstraction reaction of beta neopentylcobalamin (beta-NpCbl) in ethylene glycol-d(0) and ethylene glycol-d(4). This is the first experimental test of Klinman's hypothesis using KIE data obtained at enzyme-relevant temperatures. The key data obtained are as follows: deuterium KIEs of 23.1 +/- 3.0 at 40 degrees C to 39.0 +/- 2.3 at 10 degrees C; an activation energy difference E(D) - E(H) of 3.1 +/- 0.3 kcal mol(-)(1); and a pre-exponential factor ratio A(H)/A(D) of 0.14 +/- 0.07. Moreover, our now three sets of data (NpCbl; AdoCbl; 8-MeOAdoCbl) are shown to lie on the same ln KIE vs 1/T linear plot yielding a set of enzyme-temperature-relevant, high-precision KIE, E(D) - E(H), and A(H)/A(D) data over a relatively large, 110 degrees C temperature range. Significantly, the enzyme-free solution KIE, E(D) - E(H), and A(H)/A(D) are identical within experimental error to those for methylmalonyl-CoA mutase. This finding leads to the conclusion that there is no enzymic enhancement of the tunneling in at least this B(12)-dependent enzyme. This B(12) enzyme does, however, exploit the same (unchanged) level of tunneling measured for the nonenzymic, Ado(*) solution H(*) abstraction reaction. A discussion is presented of the still open question of if this first experimental finding, of "no enzymic enhancement of tunneling" in one B(12)-dependent enzymic system, is likely to prove more general or not. PMID- 12895107 TI - Preparation, structures, and redox and emission characteristics of the isothiocyanate complexes of hexarhenium(III) clusters [Re6(mu3-E)8(NCS)6]4- (E = S, Se). AB - Hexarhenium(III) complexes with terminal isothiocyanate ligands, [(n C(4)H(9))(4)N](4)[Re(6)(mu(3)-S)(8)(NCS)(6)] (1) and (L)(4)[Re(6)(mu(3) Se)(8)(NCS)(6)] (L(+) = PPN(+) (2a), (n-C(4)H(9))(4)N(+) (2b)), have been prepared by three different methods. Complex 1 was prepared by the reaction of [(n-C(4)H(9))(4)N](4)[Re(6)(mu(3)-S)(8)Cl(6)] with molten KSCN at 200 degrees C, while 2b was obtained by refluxing the chlorobenzene-DMF (2:1 v/v) solution of [Re(6)(mu(3)-Se)(8)(CH(3)CN)(6)](SbF(6))(2) and [(n-C(4)H(9))(4)N]SCN. The [Re(6)(mu(3)-Se)(8)(NCS)(6)](4)(-) anion was also obtained from a mixture of Cs(2)[Re(6)(mu(3)-Se)(8)Br(4)] and KSCN in C(2)H(5)OH by a mechanochemical activation at room temperature for 20 h and isolated as 2a. The X-ray structures of 1 and 2a.4DMF have been determined (1, C(70)H(144)N(10)S(14)Re(6), monoclinic, space group P2(1)/n (No. 14), a = 14.464(7) A, b = 22.059(6) A, c = 16.642(8) A, beta = 113.62(3) degrees, V = 4864(3) A(3), Z = 2; 2a.4DMF, C(162)H(144)N(14)O(4)P(8)S(6)Se(8)Re(6), triclinic, space group P1 (No. 2), a = 15.263(2) A, b = 16.429(2) A, c = 17.111(3) A, alpha = 84.07(1) degrees, beta = 84.95(1) degrees, gamma = 74.21(1) degrees, V = 4098.3(8) A(3), Z = 1). All the NCS(-) ligands in both complexes are coordinated to the metal center via nitrogen site with the Re-N distances in the range of 2.07-2.13 A. The redox potentials of the reversible Re(III)(6)/Re(III)(5)Re(IV) process in acetonitrile are +0.84 and +0.70 V vs. Ag/AgCl for [Re(6)(mu(3)-S)(8)(NCS)(6)](4)(-) and [Re(6)(mu(3) Se)(8)(NCS)(6)](4)(-), respectively, which are the most positive among the known hexarhenium complexes with six terminal anionic ligands. The complexes show strong red luminescence with the emission maxima (lambda(max)/nm), lifetimes (tau(em)/micros), and quantum yields (phi(em)) being 745 and 715, 10.4 and 11.8, and 0.091 and 0.15 for 1 and 2b, respectively, in acetonitrile. The data reasonably well fit in the energy-gap plots of other hexarhenium(III) complexes. The temperature dependence of the emission spectra and tau(em) of 1 and [(n C(4)H(9))(4)N](4)[Re(6)(mu(3)-S)(8)Cl(6)] are also reported. PMID- 12895108 TI - Synthesis, characterization, and evaluation of [Ir(ppy)2(vpy)Cl] as a polymer bound oxygen sensor. AB - This study reports new luminescent oxygen sensors in which the luminophore is covalently bound to the polymer matrix and compares their behavior to related sensors in which the luminophore is dispersed within the matrix. The cyclometalated iridium complex [Ir(ppy)(2)(vpy)Cl], 1, has been synthesized and characterized spectroscopically (absorption and emission) and by 1-D and 2-D (1)H NMR, elemental analysis, and X-ray crystallography. Complex 1 was attached via hydrosilation to hydride-terminated poly(dimethylsiloxane) (PDMS), yielding material 2. Successful luminophore attachment was determined spectroscopically from the emission properties, and through the altered physical behavior of 2 compared to a dispersion of 1 in PDMS. Hydrosilation of 1 with dimethylphenylsilane yielded [Ir(ppy)(2)(DMPSEpy)Cl], 3, which was fully characterized and used to probe the effect of hydrosilation on the spectroscopic properties of the luminophore. Evaluation of 2 as a luminescent oxygen sensor revealed significantly improved sensitivity over dispersions of 1 in PDMS. Material 2 was also blended with polystyrene (PS) to improve the physical properties of the sensor films. The blend sensors exhibited increased sensitivity relative to films of 2 alone and maintained short response times to rapid changes in air pressure. In contrast, 1 partitioned into the PS phase when dispersed in a PDMS/PS blend, resulting in longer sensor response times. PMID- 12895109 TI - Theoretical characterization of stable eta1-N2O-, eta2-N2O-, eta1-N2-, and eta2 N2-bound species: intermediates in the addition reactions of nitrogen hydrides with the pentacyanonitrosylferrate(II) ion. AB - The addition of nitrogen hydrides (hydrazine, hydroxylamine, ammonia, azide) to the pentacyanonitrosylferrate(II) ion has been analyzed by means of density functional calculations, focusing on the identification of stable intermediates along the reaction paths. Initial reversible adduct formation and further decomposition lead to the eta(1)- and eta(2)-linkage isomers of N(2)O and N(2), depending on the nucleophile. The intermediates (adducts and gas-releasing precursors) have been characterized at the B3LYP/6-31G level of theory through the calculation of their structural and spectroscopic properties, modeling the solvent by means of a continuous approach. The eta(2)-N(2)O isomer is formed at an initial stage of adduct decompositions with the hydrazine and azide adducts. Further conversion to the eta(1)-N(2)O isomer is followed by Fe-N(2)O dissociation. Only the eta(1)-N(2)O isomer is predicted for the reaction with hydroxylamine, revealing a kinetically controlled N(2)O formation. eta(1)-N(2) and eta(2)-N(2) isomers are also predicted as stable species. PMID- 12895110 TI - Tailor-made strong exchange magnetic coupling through very long bridging ligands: theoretical predictions. AB - Computational methods based on density functional theory have been applied to a prospective study of dinuclear transition metal complexes that may show strong exchange coupling interactions through very long bridging ligands. The results indicate that M(III) complexes (being M= Cr, Mn or Fe) with dicyanamidobenzene type ligands are specially promising for this purpose, since strong ferromagnetic or antiferromagnetic coupling is predicted between paramagnetic metal cations at distances as long as 25 A. The existence of ferromagnetic or antiferromagnetic coupling in the complexes with the different isomers of dicyanamidobenzene can be rationalized in terms of molecular orbitals. PMID- 12895111 TI - Effects of steric constraint on chromium(III) complexes of tetraazamacrocycles. Chemistry and excited-state behavior of 1,4-C2-cyclam complexes. AB - The synthesis and characterization of several Cr(III) complexes of the constrained macrocyclic ligand 1,4-C(2)-cyclam = 1,4,8,11 tetraazabicyclo[10.2.2]hexadecane is reported. The ligand appears to form only trans complexes, and the structure of trans-[Cr(1,4-C(2)-cyclam)Cl(2)]PF(6) is presented. The constraint imposed by the additional C(2) linkage distorts the bond angles significantly away from the ideal values of 90 and 180 degrees. The effect of the distortion is to enhance the aquation rate of trans-[Cr(1,4-C(2) cyclam)Cl(2)](+) (k(obs) for trans-[Cr(1,4-C(2)-cyclam)(H(2)O)(2)](3+) formation = 6.5 x 10(-)(2) s(-)(1), 0.01M HNO(3), 25 degrees C) by over 5 orders of magnitude relative to trans-[Cr(cyclam)Cl(2)](+). The complexes trans-[Cr(1,4 C(2)-cyclam)Cl(2)](+) and trans-[Cr(1,4-C(2)-cyclam)(CN)(2)](+) are found to have extinction coefficients four to five times higher than their cyclam analogues, owed to the lack of centrosymmetry caused by the steric constraint. The trans [Cr(1,4-C(2)-cyclam)(CN)(2)](+) complex is a very weak emitter in aqueous solution with a broad room-temperature emission centered at 735 nm (tau = 0.24 micros). Extended photolysis (350 nm, 15 h) of trans-[Cr(1,4-C(2) cyclam)(CN)(2)](+) in aqueous solution results in CN(-) ligand loss. This is in stark contrast to its unconstrained cyclam analogue, which is photoinert and has a room-temperature emission lifetime of 335 micros. PMID- 12895112 TI - Enantioselective recognition between chiral alpha-hydroxy-carboxylates and macrocyclic heptadentate lanthanide(III) chelates. AB - Three novel heptacoordinated Ln(III) complexes (Ln = Gd and Yb) have been synthesized and investigated by (1)H NMR spectroscopy. These complexes contain two stereogenic centers, one associated with a deltadeltadeltadelta or lambdalambdalambdalambda conformation of the ethylenediamine moieties in the tetraazamacrocycle and the latter arises from the orientation (Delta or Lambda) of the coordinating arms. Evidence has been gained for the occurrence of a fast exchange between all the possible conformers. Upon addition of several (S)-alpha hydroxy-carboxylate substrates, the formation of stable ternary adducts has been obtained. Their (1)H NMR spectra are consistent with the presence of two diastereoisomers differing in the conformation adopted by the macrocyclic ligand wrapping the lanthanide(III) ion. The interaction leading to the formation of the ternary complexes is enantioselective depending on the hydrophilicity of the alpha-hydroxy-carboxylate. PMID- 12895113 TI - Synthesis, structure, and properties of the new intermetallic compounds SrPdTl2 and SrPtTl2. AB - The title compounds have been synthesized and characterized structurally and through property measurements and electronic structure calculations. Single crystal X-ray diffraction analyses reveal that the two compounds crystallize in an orthorhombic system, MgCuAl(2) type (Cmcm, Z = 4, a = 4.486(2), 4.491(3) A, b = 10.991(5), 10.990(6) A, c = 8.154(1), 8.140(4) A for SrPdTl(2), and SrPtTl(2), respectively). The structure can be directly derived from that of hexagonal SrTl(2) (CaIn(2) type) in which four-bonded thallium atoms in shared puckered hexagons generate tunnels. The Pd or Pt is encapsulated (with symmetry reduction) on the side of each tunnel within a distorted trigonal prism. Band structure calculations (EHTB) on both SrTl(2) and SrPdTl(2) demonstrate the effects of the conversion, with strong Pd-Tl bonding and appreciable electron transfer from Tl to Pd. Property measurements show that SrPdTl(2) is metallic, as expected. PMID- 12895114 TI - Spectroscopic and density functional studies of the dinitrosyl metalloporphyrin complexes Fe(P)(NO)2 and Ru(P)(NO)2. AB - Experimental evidence including infrared spectra for the formation of the dinitrosyl metalloporphyrin complexes M(P)(NO)(2) (M = Ru or Fe, P = tetraphenylporphyrin (TPP), octaethylporphyrin (OEP), or tetra-m-tolylporphryin (TmTP)) is described. Although observation of a single NO stretching band in the IR spectrum of each M(P)(NO)(2) complex first suggested a centrosymmetric (D(4)(h)() or C(2)(h)()) structure, DFT geometry optimizations and frequency calculations of model complexes indicate that the trans-syn (C(2)(v)()) conformation should be more stable. The frequency calculations resolve the apparent ambiguity in the IR spectra in terms of the relative oscillator strengths of the predicted IR bands. PMID- 12895115 TI - Stoichiometric and catalytic activation of the alpha- and beta-2,3,4-tri-O-acetyl 5-thioxylopyranosyl bromide inside the cavity of the Pd3(dppm)3(CO)2+ cluster. AB - The title cluster (Pd(3)(2+)) exhibits a pronounced affinity for Br(-) ions to form the very stable Pd(3)(Br)(+) adduct. Upon a 2-electron reduction, a dissociative process occurs generating Pd(3)(0) and eliminating Br(-) according to an ECE mechanism (electrochemical, chemical, electrochemical). At a lower temperature (i.e. -20 degrees C), both ECE and EEC processes operate. This cluster also activates the C-Br bond, and this work deals with the reactivity of Pd(3)(2+) with 2,3,4-tri-O-acetyl-5-thioxylopyranosyl bromide (Xyl-Br), both alpha- and beta-isomers. The observed inorganic product is Pd(3)(Br)(+) again, and it is formed according to an associative mechanism involving Pd(3)(2+).Xyl-Br host-guest assemblies. In an attempt to render the C-Br bond activation catalytic, these species are investigated under reduction conditions at two potentials (-0.9 and -1.25 V vs SCE). In the former case, the major product is Xyl-H, issued from a radical intermediate Xyl(*) abstracting an H atom from the solvent. Evidence for Xyl(*) is provided by the trapping with TEMPO (2,2,6,6 tetramethyl-1-piperidinyloxy) and DMPO (5,5'-dimethylpyrroline-N-oxyde). In the second case, only one product is observed, 3,4-di-O-acetyl-5-thioxylal, which is issued from the Xyl(-)() intermediate anion. PMID- 12895116 TI - X-ray structures, photophysical characterization, and computational analysis of geometrically constrained copper(I)--phenanthroline complexes. AB - A series of three geometrically constrained C(2)-symmetric Cu(I) mono phenanthroline complexes were characterized by X-ray structural analysis, and their photophysical properties were investigated by absorption and emission spectroscopy. Visible light excitation yielded metal-to-ligand charge-transfer (MLCT) excited states with luminescence lifetimes up to 155 ns. Ultrafast transient absorption spectroscopy provided further insights into the excited state dynamics and suggests for all three complexes the formation of a phenanthroline radical anion. In agreement with electrochemical measurements, the data further indicate that coordinative rearrangements are involved in nonradiative deactivation of the excited states. According to time-dependent density functional theory calculations (B3LYP/6-31G), the major MLCT transitions are polarized along the C(2) axis of the complex and originate predominantly from the copper d(xz) orbital. The computational analysis identifies an excited-state manifold with a number of close-lying, potentially emissive triplet states and is in agreement with the multiexponential decay kinetics of the MLCT luminescence. The relationship between structural and photophysical data of the studied Cu(I) mono-phenanthroline complexes agrees well with current models describing the photophysics of the related Cu(I) bis-diimine complexes. PMID- 12895117 TI - Toward optimized high-relaxivity MRI agents: the effect of ligand basicity on the thermodynamic stability of hexadentate hydroxypyridonate/catecholate gadolinium(III) complexes. AB - The thermodynamic stabilities of the Gd(III) complexes of five hexadentate ligands, which incorporate the 2,3-dihydroxyterephthalamide and 2,3 hydroxypyridonate chelating moieties, have been determined by potentiometric and spectrophotometric titration. The ligands were chosen to span a range of basicities while maintaining a similar tripodal structural motif, facilitating a study of the effect of ligand basicity on the thermodynamic stability of the Gd(III) complexes. The relative stability of the five complexes is found to be highly pH dependent, with the most acidic ligands forming the most stable complexes at low pH and more basic ligands forming more stable complexes at high pH. The most stable Gd(III) complex at a physiological pH of 7.4 is formed with a ligand of intermediate basicity and is of stability comparable to that of Gd(III) complexes that feature eight-coordinate amino-carboxylate ligands and are currently used as magnetic resonance imaging contrast agents in diagnostic medicine. A single-crystal X-ray structure of the intermediate compound 3-hydroxy 6-methyl-2-oxo-1,2-dihydropyridine-4-carboxylic acid ethyl ester is described: This compound crystallizes in the triclinic space group P1 with a = 7.4801(3) A, b = 8.0671(3) A, c = 8.3457(4) A, alpha = 72.242(2) degrees, beta = 80.693(2) degrees, gamma = 69.943(3) degrees, V = 449.60(3) A(3), Z = 2, and R = 0.042. PMID- 12895118 TI - Crystal structures and vibrational and solution and solid-state (CPMAS) NMR spectroscopic studies in triphenyl phosphine, arsine, and stibine silver(I) bromate systems, (R3E)xAgBrO3 (E = P, As, Sb; x = 1-4). AB - Adducts of triphenyl phosphine, triphenyl arsine, and triphenyl stibine with silver(I) bromate have been synthesized and characterized both in solution ((1)H and ESI MS spectroscopy) and in the solid state (IR, single-crystal X-ray structure analysis). The triphenyl phosphine complexes have been also investigated by (31)P[(1)H] solution and (31)P cross-polarization magic-angle spinning (CPMAS) NMR spectroscopy. The topology of the structures in the solid state was found to depend on the nature of EPh(3) and on the stoichiometric ratio AgBrO(3)/EPh(3). In AgBrO(3)/PPh(3) (1:1)(4) (1) and AgBrO(3)/PPh(3) (1:2) (2), the bromate is in the unfamiliar and hitherto structurally uncharacterized role of coordinating ligand, the complex having a mononuclear form in 2 and a less familiar tetrameric form in 1. In AgBrO(3)/AsPh(3) (1:4).CH(3)OH (7) and AgBrO(3)/SbPh(3) (1:4).C(2)H(5)OH (11), the cations are the familiar homoleptic [Ag(EPh(3))(4)](+) array with the bromate role simply that of counterion. The AgBrO(3)/AsPh(3) (1:2)(2).0.7"H(2)O" derivative (6) is binuclear L(2)Ag(mu BrO(3))(2)AgL(2) with a four-membered ring core (L = AsPh(3)). PMID- 12895119 TI - Chalcogeno-urea ligands on a phosphadiazonium Lewis acceptor: a new synthetic approach to Ch-P bonds (Ch = O, S, Se). AB - The isolation and characterization of the first intermolecular chalcogeno-urea complexes of iminophosphines are described. Trifluoromethylsulfonyloxy(2,4,6-tri tert-butylphenylimino)phosphine, MesNPOTf, reacts quantitatively with chalcogenoimidazolines (ChIm, Ch = O, S, Se) and 1,3-dimethyldiphenylurea (OU) to give Lewis acid-base complexes, [MesNP.ChIm]OTf and [MesNP.OU]OTf. Single crystal X-ray diffraction studies indicate that the closest contact of the chalcogeno urea donor occurs at phosphorus in all cases, representing compounds that contain examples of O-P, S-P, and Se-P coordinate bonds. In all complexes, coordination of the ligand causes significant displacement of the OTf anion, and the resulting cations [MesNP.L](+) are best described as complexes of a neutral ligand on a phosphadiazonium Lewis acceptor. As such, the complex ions [MesNP.L](+) are novel examples of cationic systems containing dicoordinate phosphorus centers. The complexes highlight the potential for electron-rich centers to behave as Lewis acids despite the presence of a lone pair of electrons at the acceptor site. PMID- 12895120 TI - Metal binding characteristics of a laterally nonsymmetric aza cryptand upon functionalization with a pi-acceptor group. AB - The laterally nonsymmetric aza cryptand synthesized by condensing tris(2 aminoethyl)amine (tren) with tris[[2-(3-(oxomethyl)phenyl)oxy]ethyl]amine readily forms mononuclear inclusion complexes with both transition and main-group metal ions. In these complexes, the metal ion occupies the tren-end of the cavity making bonds with the three secondary amino and the bridgehead N atoms. When a strong pi-acceptor group such as 2,4-dinitrobenzene is attached to one of the secondary amines, the binding property of the cryptand changes drastically. When perchlorate or tetrafluoroborate salts of Ni(II), Cu(II), Zn(II), or Cd(II) are used, the metal ion enters the cavity which can be monitored by the hypsochromic shift of the intramolecular charge-transfer transition from the donor amino N atom to the acceptor dinitrobenzene. However, in the presence of coordinating ions such as Cl(-), N(3)(-), and SCN(-), the metal ion comes out of the cavity and binds the cryptand outside the cavity at a site away from the dinitrobenzene moiety. Four such complexes are characterized by X-ray crystallography. Thus, a metal ion can translocate between inside and outside of the cryptand cavity depending upon the nature of the counter anion. PMID- 12895121 TI - Group 6 imido complexes supported by diamido-donor ligands. AB - Reactions of the lithiated diamido-pyridine or diamido-amine ligands Li(2)N(2)N(py) or Li(2)N(2)N(am) with [W(NAr)Cl(4)(THF)] (Ar = Ph or 2,6 C(6)H(3)Me(2); THF = tetrahydrofuran) afforded the corresponding imido-dichloride complexes [W(NAr)(N(2)N(py))Cl(2)] (R = Ph, 1, or 2,6-C(6)H(3)Me(2), 2) or [W(NAr)(N(2)N(am))Cl(2)] (R = Ph, 3, or 2,6-C(6)H(3)Me(2), 4), respectively, where N(2)N(py) = MeC(2-C(5)H(4)N)(CH(2)NSiMe(3))(2) and N(2)N(am) = Me(3)SiN(CH(2)CH(2)NSiMe(3))(2). Subsequent reactions of 1 with MeMgBr or PhMgCl afforded the dimethyl or diphenyl complexes [W(NPh)(N(2)N(py))R(2)] (R = Me, 5, or Ph, 6), respectively, which have both been characterized by single crystal X ray diffraction. Reactions of Li(2)N(2)N(py) or Li(2)N(2)N(am) with [Mo(NR)(2)Cl(2)(DME)] (R = (t)Bu or Ph; DME = 1,2-dimethoxyethane) afforded the corresponding bis(imido) complexes [Mo(NR)(2)(N(2)N(py))] (R = (t)Bu, 7, or Ph, 8) and [Mo(N(t)Bu)(2)(N(2)N(am))] (9). PMID- 12895122 TI - Neutral and ionic aluminum, gallium, and indium compounds carrying two or three terminal ethynyl groups. AB - The syntheses of the ionic compounds [Li(+).2 dioxane (2,6 iPr(2)C(6)H(3)N(SiMe(3))Al(C triplebond CSiMe(3))(3))(-)].0.75 dioxane (1), [(Li(+))(2).(dioxane)(7)](0.5) [2,6-iPr(2)C(6)H(3)N(SiMe(3))Ga(C triplebond CSiMe(3))(3)(-)].1.5 dioxane (2), and [(Li(+))(2).(dioxane)(7)](0.5) [2,6 iPr(2)C(6)H(3)N(SiMe(3))In(C triplebond CSiMe(3))(3)(-)].1.5 dioxane (3) by the reaction of the corresponding organo metal chloride with LiC triplebond CSiMe(3) are reported. The neutral ethynyl compounds Br-Al(C triplebond CtBu)(2).2 THF (4), Cl-Ga(C triplebond CtBu)(2).THF (5), Cl-In(C triplebond CtBu)(2).2 THF (6), Al(C triplebond CtBu)(3).C[N(Me)CMe](2) (7), Ga(C triplebond CtBu)(3).dioxane (8), and In(C triplebond CtBu)(3).NEt(3) (9) have been obtained in good yields from the reaction of AlBr(3), GaCl(3), and InCl(3) with LiC triplebond CtBu in the presence of a Lewis base. Compound 7 is the first heterocyclic carbene substituted ethynyl derivative. Aluminum and gallium compounds with three terminal ethynyl groups Al(C triplebond CPh)(3).NMe(3) (10) and Ga(C triplebond CPh)(3).NMe(3) (11) have been prepared by the reaction of AlH(3).NMe(3) or GaH(3).NMe(3) with three equivalents of phenylethyne. All the above-mentioned compounds have been structurally studied. In compound 1 the lithium ion is coordinated to the three terminal ethynyl groups, whereas in compounds 2 and 3 the lithium is coordinated to the solvent (dioxane). Compound 8 crystallizes as a coordination polymer with dioxane molecules bridging the individual gallium units. PMID- 12895123 TI - Molecular structures, bond energies, and bonding analysis of group 11 cyanides TM(CN) and isocyanides TM(NC) (TM = Cu, Ag, Au). AB - We report on quantum chemical calculations at the DFT (BP86/TZP) and ab initio (CCSD(T)/III+) levels of the title compounds. The geometries, vibrational spectra, heats of formation, and homolytic and heterolytic bond dissociation energies are given. The calculated bond length of Cu-CN is in reasonable agreement with experiment. The theoretical geometries for CuNC and the other group 11 cyanides and isocyanides which have not been measured as isolated species provide a good estimate for the exact values. The theoretical bond dissociation energies and heats of formation should be accurate with an error limit of +/-5 kcal/mol. The calculation of the vibrational spectra shows that the C-N stretching mode of the cyanides, which lies between 2170 and 2180 cm(-)(1), is IR inactive. The omega(1)(C-N) vibrations of the isocyanides are shifted by approximately 100 cm(-)(1) to lower wavenumbers. They are predicted to have a very large IR intensity. The nature of the metal-ligand interactions was investigated with the help of an energy partitioning analysis in two different ways using the charged fragments TM(+) + CN(-) (TM = transition metal) and the neutral fragments TM(*) + CN(*) as bonding partners. The calculations suggest that covalent interactions are the driving force for the formation of the TM-CN and TM-NC bonds, but the finally formed bonds are better described in terms of interactions between TM(+) and CN(-), which have between 73% and 80% electrostatic character. The contribution of the pi bonding is rather small. The lower energy of the metal cyanides than that of the isocyanides comes from the stronger electrostatic interaction between the more diffuse electron density at the carbon atom of the cyano ligand and the positively charged nucleus of the metal. PMID- 12895124 TI - High-dimensional architectures from the self-assembly of lanthanide ions with benzenedicarboxylates and 1,10-phenanthroline. AB - Six new coordination polymers, [Eu(1,2-BDC)(1,2-HBDC)(phen)(H(2)O)](n) (1), [Eu(2)(1,3-BDC)(3)(phen)(2)(H(2)O)(2)](n).4nH(2)O (2), [Eu(1,4 BDC)(3/2)(phen)(H(2)O)](n) (3), [Yb(2)(1,2-BDC)(3)(phen)(H(2)O)(2)](n).3.5nH(2)O (4), [Yb(2)(1,3-BDC)(3)(phen)(1/2)](n) (5), and [Yb(2)(1,4 BDC)(3)(phen)(2)(H(2)O)](n) (6), were synthesized by hydrothermal reactions of lanthanide chlorides with three isomers of benzenedicarboxylic acid (H(2)BDC) and 1,10-phenanthroline (phen), and characterized by single-crystal X-ray diffraction. 1 has a 2-D herringbone architecture with a Z-shaped cavity. 2 and 5 have different 3-D networks, but both are formed by 1,3-BDC anions bridging metal centers (Eu or Yb) via carboxylate groups. 3 and 6 possess similar layer structures which are further constructed to form 3-D networks by hydrogen bonds and/or pi-pi aromatic interactions. 4 comprises 1-D chains that are further interlinked via hydrogen bonds, resulting in a 3-D network. In the three europium complexes, all the europium ions are eight-coordinated, while the coordination numbers of the ytterbium ions in other three-coordination polymers range from six to eight. Crystal data: for 1, monoclinic, space group P2(1)/c, with a = 12.565(6) A, b = 16.005(8) A, c = 12.891(6) A, beta = 102.173(8) degrees, and Z = 4; for 2, monoclinic, space group P2(1)/c, with a = 20.979(4) A, b = 11.5989(19) A, c = 20.810(3) A, beta = 110.391(3) degrees, and Z = 4; for 3, triclinic, space group P1, with a = 10.331(5) A, b = 10.887(5) A, c = 11.404(5) A, alpha = 107.660(7) degrees, beta = 91.787(7) degrees, gamma = 112.946(6) degrees, and Z = 2; for 4, triclinic, space group P1, with a = 11.517(5) A, b = 13.339(5) A, c = 13.595(6) A, alpha = 87.888(7) degrees, beta = 67.759(6) degrees, gamma = 68.070(6) degrees, and Z = 2; for 5, orthorhombic, space group C222(1), with a = 8.174(2) A, b = 24.497(7) A, c = 29.161(8) A, and Z = 8; for 6, triclinic, space group P1, with a = 10.349(3) A, b = 11.052(3) A, c = 19.431(6) A, alpha = 105.464(4) degrees, beta = 91.300(5) degrees, gamma = 93.655(5) degrees, and Z = 2. The magnetic properties of 1 and 4 were investigated. The photophysical properties of 1 were also studied. PMID- 12895125 TI - Novel Pb(II), Zn(II), and Cd(II) coordination polymers constructed from ferrocenyl-substituted carboxylate and bipyridine-based ligands. AB - Treatment of two kinds of ferrocenyl-substituted carboxylate ligands (3 ferrocenyl-2-crotonic acid, HOOC-CH=(CH(3))CFc (Fc=(eta(5)-C(5)H(5))Fe(eta(5) C(5)H(4))) or O-ferrocecarbonyl benzoic acid, o-HOOCC(6)H(4)COFc with Pb(OAc)(2).3H(2)O, Zn(OAc)(2).2H(2)O, or Cd(OAc)(2).2H(2)O) resulted in four novel ferrocene-containing coordination polymers [[Pb(mu(2)-eta(2) OOCCH=(CH(3))CFc)(2)].MeOH](n) (1), [[Zn(o-OOCC(6)H(4)COFc)(2)(4,4' bipy)(H(2)O)(2)].2MeOH.2H(2)O](n) (4,4'-bipy = 4,4'-bipyridine) (2), [[Cd(o OOCC(6)H(4)COFc)(2)(bpe)(MeOH)(2)].2H(2)O](n) (bpe = 1,2-bis(4-pyridyl)ethene) (3), and [Pb(o-OOCC(6)H(4)COFc)(eta(2)-o-OOCC(6)H(4)COFc)(bpe)](n)() (4). Their crystal structures have been characterized by single X-ray determinations. In polymer 1, Pb(II) ions are bridged by tridentate FcC(CH(3))=CHCOO(-) anions, forming an infinite chain [Pb(mu(2)-eta(2)-OOC=CH(CH(3))CFc)(2)](n). In polymers 2-4, there are three kinds of components, metal ions, o-FcCOC(6)H(4)COO(-) units, and organic bridging ligands. The bipyridine-based ligands connect metal ions leading to a one-dimensional chain with o-FcCOC(6)H(4)COO(-) units acting as monodentate or chelate ligands in the side chain. Such coordination polymers containing ferrocenyl-substituted carboxylate and bipyridine-based ligands are very rare. The solution-state differential pulse voltammetries of polymers 1-4 were determined. The results indicate that the half-wave potential of the ferrocenyl moieties is influenced by the Pb(II) ions in polymer 1 and strongly influenced by Zn(II), Cd(II), or Pb(II) ions in polymers 2-4. The thermal properties of the four polymers were also investigated. PMID- 12895126 TI - Controlled reactions on a copper surface: synthesis and characterization of nanostructured copper compound films. AB - We report the synthesis of nanostructured copper compound films on a copper surface under mild conditions. A series of low-dimensional structures including Cu(OH)(2) fibers and scrolls, CuO sheets and whiskers, and Cu(2)(OH)(2)CO(3) rods have been successfully grown on the copper surfaces at ambient temperature and pressure. Most of the structures are phase-pure single crystallites. The films were formed by the direct oxidation of copper in aqueous solutions of NaOH with an oxidant (NH(4))(2)S(2)O(8). The evolution of the ultrafine structures as a function of the reaction conditions has been revealed, from fibers of Cu(OH)(2) to scrolls of Cu(OH)(2) to sheets or whiskers of CuO. By replacing NaOH with NaHCO(3) in the synthesis, square/rectangular rod arrays of Cu(2)(OH)(2)CO(3) were obtained. The controlled reactions allow the large-scale, template-free, cost-effective synthesis of copper compound films with ordered, uniform, stable, ultrafine structures. PMID- 12895127 TI - Investigation of the interaction between 1-hydroxyethane-1,1'-diphosphonic acid (HEDP) and uranium(VI). AB - A detailed study, using a panel of spectroscopic analytical methods, of the complexation between 1-hydroxyethane-1,1'-diphosphonic acid (HEDP) and uranyl ion (UO(2)(2+)) is reported. Results suggest that the metal complex is present as only 1:1 (metal/ligand) species at low concentration (<10(-)(4) M). The conditional constants of this complex were determined at various pH using time resolved laser-induced fluorescence (TRLIF) and electrospray ionization mass spectrometry (ESI-MS). Further investigations indicate the presence of a 1:2 (metal/ligand) complex at higher concentrations ( approximately 10(-)(2) M). Selectivity studies as well as structural aspects are presented. PMID- 12895128 TI - Facile preparation and photophysics of near-infrared luminescent lanthanide(III) monoporphyrinate complexes. AB - The synthesis, characterization, and single crystal X-ray diffraction structures of a series of monoporphyrinate, trivalent lanthanide complexes with the monoanionic ligands hydridotris(1-pyrazolyl)borate (Tp) and (cyclopentadienyl)tris(diethylphosphinito)cobaltate (L(OEt)) having the general formulas M(TPP)(L) (M = Yb, Tm, Er, Ho, Nd, Pr; TPP = 5,10,15,20 tetraphenylporphyrinate; L = Tp, L(OEt)) are described. The photophysical properties of these complexes are also presented including their absorption, emission, and transient absorption properties. PMID- 12895130 TI - Assessment and interpretation of isokinetic muscle strength during growth and maturation. AB - The majority of strength studies examining changes during growth and maturation have investigated isometric actions, which tell us little about the muscle under dynamic conditions. There are numerous methodological issues in the isokinetic testing of paediatric populations that require further investigation. However, several studies have indicated that children can be reliably assessed isokinetically using both concentric and eccentric actions. Most paediatric studies have examined the knee joint and more data are needed to elucidate the reliability of upper body isokinetic strength testing. The age- and sex associated development of isokinetic strength is less well understood. Studies have indicated that isokinetic strength increases with age but the mechanisms associated with this increase require further investigation. Current data are also conflicting regarding the age at which sex differences become apparent in isokinetic strength. More work is needed to examine the influence of maturation on isokinetic strength development, but available data suggest that maturation is a non-significant contributory factor once stature and body mass are accounted for. Most studies have demonstrated a significant relationship between stature, body mass and isokinetic strength during growth and maturation. The importance that changes in body composition during growth have on isokinetic strength has been investigated using fat-free mass and muscle cross-sectional area. Data have shown that although fat-free mass and muscle cross-sectional area are important contributors to isokinetic strength, other unexplained factors also influence isokinetic strength development. Additional work needs to investigate possible qualitative changes in muscle during growth and maturation. More work is also needed to examine changes in eccentric strength with age and to investigate sex differences in upper body isokinetic strength. Future studies should preferably be longitudinal in nature and examine known covariates simultaneously using appropriate statistical techniques. PMID- 12895129 TI - Abdominal adipose tissue distribution and metabolic risk. AB - It is now established that waist circumference remains a significant predictor of disease and metabolic risk independent of obesity measured by body mass index. These observations are reinforced by a large body of evidence implicating abdominal obesity measured by imaging modalities in the pathogenesis of numerous metabolic risk factors. However, the extent to which abdominal subcutaneous or visceral adipose tissue (AT) independently contribute to the established association between abdominal obesity and metabolic risk remains a subject of considerable research. Discrepancies in the literature may be partially explained by methodological issues, as currently there is no accepted definition for either visceral or abdominal subcutaneous AT. Accordingly, there is confusion concerning how best to measure these depots in metabolic and descriptive studies. Further, despite numerous studies linking abdominal AT depots with metabolic risk factors, the underlying mechanisms remain unclear. The primary purpose of this review is to examine the independent contribution of visceral and abdominal subcutaneous AT to the aetiology of obesity-related health risk. We begin by considering specific methodological issues with respect to the current classification of abdominal AT as measured by imaging methods. When necessary, we present original data to reinforce important concepts not suitably addressed in the literature. We conclude with a consideration of proposed mechanisms that may link abdominal adiposity and metabolic risk. PMID- 12895131 TI - Epidemiology of non-submersion injuries in aquatic sporting and recreational activities. AB - Although the issues of drowning and near-drowning in aquatic sporting and recreational activities receive considerable attention in the epidemiological literature, there is not a recognised literature on non-submersion injuries occurring in these activities. This review draws together the epidemiological literature on non-submersion injuries and describes the incidence, nature and causes of these injuries, common risk factors, and strategies for prevention. Activities covered by the review include swimming, diving, boating, surf sports, fishing, water polo and water sliding. For most activities there is a dearth of good quality descriptive studies, with most involving cases-series designs and few providing estimates of incidence. Inconsistencies in inclusion criteria and the reporting of incidence rates makes comparisons within and between activities difficult. Incidence rates were identified for most activities and in general the incidence of injury was low, especially for more serious injury. However, some activities were associated with severely disabling injury, such as spinal cord injury (diving) and amputation (from propeller strikes in water skiing and swimming). Only three studies reporting the significance of postulated risk factors were identified. Lack of knowledge about the water being entered and alcohol consumption are significant risk factors in recreational diving; increased blood alcohol concentrations were reported to increase the risk of death in boating; and obesity and tandem riding were reported to increase the risk of injury on public water slides. Few evaluations of preventive measures were identified. Two studies reported reductions in the incidence of water slide injuries following the introduction of design changes and supervision, but neither had a non-intervention comparison group. Improvements in swimming and diving skills were reported in three studies, but these were not designed to measure changes in the risk of injury.This review demonstrates that there is a need for well-designed epidemiological research on non-submersion injury in aquatic sporting and recreational activities. The first priority should be for studies designed to describe accurately the incidence, nature, severity and circumstances of these injuries, followed by research on the significance of postulated risk factors. Once this research has been undertaken, interventions targeted at reducing the incidence of injury in aquatic sporting and recreational activities can be designed and evaluated. PMID- 12895132 TI - Elucidating the unexplained underperformance syndrome in endurance athletes : the interleukin-6 hypothesis. AB - The unexplained underperformance syndrome (UPS), previously known as the overtraining syndrome (OTS), has been defined as a persistent decrement in athletic performance capacity despite 2 weeks of relative rest. It has been proposed that UPS may be caused by excessive cytokine release during and following exercise causing a chronic inflammatory state and 'cytokine sickness'. This article extends that hypothesis by proposing that time-dependent sensitisation could provide a model through which the aetiology of UPS may be explained. In this model, the principal abnormal factors in UPS are an increased production of and/or intolerance to interleukin (IL)-6 during exercise. Strategies to attenuate the IL-6 response to exercise that may also reduce an athlete's susceptibility to UPS are proposed. PMID- 12895133 TI - Pediatric molluscum contagiosum: optimal treatment strategies. AB - Pediatric molluscum contagiosum virus (MCV) is a common pox viridae infection that represents a common public health issue. The spread of the virus among children is rapid and easy. The virus produces a number of substances that block immune response formation in the infected host. Despite the benign and self limited nature of the condition, one-third of children have symptoms from, or secondary reactions to the infection, including pruritus, erythema and, occasionally, inflammation and pain. Patients with pruritus autoinoculate the virus through scratching, thereby exacerbating their conditions. While adults cope well with unanesthetized curettage of lesions, children require less painful therapeutic options. The options for therapy are manifold. Therapy should begin with gentle skin care and antipruritics to prevent symptoms, and to prevent the spread of the disease. Therapies with good efficacy and low risk of pain for the patient include in-office usage of cantharidin and the use of local anesthetics, such as topical lidocaine (lignocaine) preparations in combination with the curettage of visible lesions. Alternatively, cryosurgery can be performed to eradicate lesions in-office. At-home therapeutics are often preferred by parents and children, and include imiquimod, retinoids, and alpha-hydroxy acids. Although a variety of such at-home therapies are available, none are as effective or as rapid acting as in-office therapy. Further research in large clinical trials is required to increase knowledge on prevention, optimal treatment, and long-term outcome with this disease. PMID- 12895134 TI - Coronary artery vasculopathy in pediatric cardiac transplant patients: the therapeutic potential of immunomodulators. AB - The single largest cause of late graft loss in pediatric cardiac transplantation is transplant coronary artery vasculopathy (CAV). The mechanism of CAV remains unknown; it appears to have both immune and non-immune causes. The final common pathway of these mechanisms is endothelial activation, a prothrombotic environment, and endothelial damage with subsequent diffuse intimal proliferation. The disease process has largely been thought to be progressive and unresponsive to treatment. Re-transplantation has been advocated as the only definitive treatment. The appropriate management is largely unknown; intervention or surgical management has had limited utility, while medical management appears to have the most promise. Improvement in outcome can be achieved by optimizing non-immune factors and aggressive management of the immune mechanisms. Long-term survival of transplant patients after diagnosis with CAV is now being reported. PMID- 12895135 TI - New and emerging therapies for short bowel syndrome in children. AB - This review provides an overview of traditional as well as emerging therapies useful in the management of pediatric short bowel syndrome. Pediatric short bowel syndrome is relatively uncommon; however, when it does occur, it presents a unique challenge to medical care providers. The use of parenteral and enteral nutrition to maximize growth and enhance intestinal adaptation so as to increase absorptive surface area has been the primary focus of therapy. In recent years, the advent of pharmacologic advances, including the use of antibacterial drugs, anti-motility drugs and hormonal therapies, has had a significant impact on this condition. At times, surgery may be indicated for dealing with complications, or providing alternative therapy such as transplantation. With ongoing research, it is likely that improved pharmacologic therapy will be available for enhanced intestinal adaptation, control of gut motility, treatment of small bowel bacterial overgrowth, and treatment of rejection following small intestinal transplantation. PMID- 12895136 TI - Childhood ependymoma: a systematic review of treatment options and strategies. AB - Childhood intracranial ependymoma have a dismal prognosis, especially in young children and when a gross total resection cannot be performed. Even in the absence of a radiologically proven residuum, around two-thirds of these young children will have a recurrence. Adjuvant therapy is therefore necessary for most, if not all, patients. Despite some indication that benign ependymoma (WHO grade II) could show a better outcome, histology cannot be used at present to stratify treatment protocols.Craniospinal irradiation combined with posterior fossa boost has deleterious adverse effects on cognition. Consequently, pediatric oncology teams have, firstly, tried to use chemotherapy to delay or avoid irradiation, and secondly, progressively reduced irradiation fields to the tumor bed without altering the prognosis. Cisplatin, at a dose of 120 mg/m(2) (cumulated response rate of 34% [95% CI 19-54%]) is the only single agent that has reproducibly shown some efficacy in ependymoma. Despite some combinations showing efficacy in the adjuvant setting, childhood intracranial ependymomas can, in general, be considered as chemoresistant. The overexpression of the multidrug resistance-1 gene and the 06-methylguanine-DNA methyltransferase have been implicated as possible mechanisms for this phenomenon. As the use of chemotherapy with current agents is questionable, phase II studies with new agents and combinations are necessary. Since the main problem of this disease is local relapse, it may not be necessary to irradiate the whole posterior fossa. However, local control of the disease by irradiation has to be improved. In this respect, hyperfractionation or radiosensitizers may be valuable therapeutic options. The treatment of children with ependymoma is a challenge for all caregivers. There is no doubt that any possible improvement in the management of this rare tumor will only be the result of well designed cooperative trials. PMID- 12895137 TI - Comparative efficacy of two once daily methylphenidate formulations (Ritalin LA and Concerta) and placebo in children with attention deficit hyperactivity disorder across the school day. AB - OBJECTIVES: The primary objective was to compare the differences in clinical efficacy of the starting dose of Ritalin LA (20mg) to the starting dose of Concerta (18mg), in a laboratory school setting for the duration of an entire school day. Secondary objectives were to compare Ritalin LA 20mg with Concerta 36mg, and Ritalin LA and both Concerta doses versus placebo across the school day. METHODS: Thirty-six children (29 males, 7 females), aged 6-12 years, with attention deficit hyperactivity disorder, previously stabilized on methylphenidate (MPH), completed this four-way, randomized, single-blind crossover, analog classroom study. Patients were evaluated on day 0 and randomized to receive treatment on days 7, 14, 21, and 28 (Ritalin LA 20mg, Concerta 18mg, Concerta 36mg, or placebo). RESULTS: Swanson, Kotkin, Agler, M Flynn and Pelham Rating Scale (SKAMP)-attention: The effect of Ritalin LA 20mg across the morning was statistically different from that of Concerta 18mg and 36mg, as demonstrated by the change in the area under the curve (AUC) during the first 4 hours (0-4) from pre-dose. AUC((0-4)) for RitalinLA was -2.48 versus 1.36 for Concerta 18mg (p = 0.015), and -1.55 for Concerta 36mg (p = 0.043). AUC((0-8)) change from pre-dose for Ritalin LA was -4.48 versus -2.72 for Concerta 18mg (p = 0.074), and -3.24 for Concerta 36mg (p = 0.208).SKAMP deportment: AUC((0-4)) for Ritalin LA was -1.67 compared with -0.28 for Concerta 18mg (p < 0.001), and -0.55 for Concerta 36mg (p = 0.004). AUC((0-8)) change from pre-dose for Ritalin LA was -2.81 compared with -0.82 for Concerta 18mg (p = 0.018), and -1.34 for Concerta 36mg (p = 0.078).Combined: Mean AUC((0-4)) change from pre-dose for Ritalin LA was -2.05 compared with -0.78 for Concerta 18mg (p < 0.001), -1.01 for Concerta 36mg (p = 0.003). The mean AUC((0-8)) change from pre dose for Ritalin LA was -3.58 compared with -1.70 for Concerta 18mg (p = 0.010), 2.22 for Concerta 36mg (p = 0.061). Math test-attempted: Mean pre-dose score for Ritalin LA was about 73 compared with 74, 90, and 81 for Concerta 18mg, 36mg, and placebo, respectively. Mean AUC((0-8)) change from pre-dose for Ritalin LA was 202 compared with 115 for Concerta 18mg (p = 0.135), 137 for Concerta 36mg (p = 0.265). Math test-correct: Mean pre-dose score for Ritalin LA was 68 compared with 64, 78, and 76 for Concerta 18mg, 36mg, and placebo, respectively. Mean AUC((0-8)) change from pre-dose for Ritalin LA was 183 compared with 100 for Concerta 18mg (p = 0.144), and 117 for Concerta 36mg (p = 0.245). SAFETY: One patient from each treatment group experienced a single mild adverse event that included abdominal pain, nausea, and dyspnea. CONCLUSION: While both Ritalin LA and Concerta were shown to be effective, the different release profiles of each formulation can result in distinct differences between the effects on measures of attention and deportment. PMID- 12895139 TI - Blood pressure variability and cardiovascular control mechanisms in hypertension. AB - Blood pressure variability is the result of a complex interaction between external stimulation and several cardiovascular control mechanisms and is enhanced in all those conditions characterized by an impaired autonomic regulation of circulation. In particular, blood pressure variability is enhanced in hypertension over 24 h and may independently contribute to the cardiovascular complications of this condition. The paper by Mussalo and co-workers in this issue of Clinical Science offers additional information in this field by suggesting that different forms of hypertension are characterized by different patterns of blood pressure variability, and that these differences may reflect variations in the underlying autonomic regulation of the heart and blood vessels. PMID- 12895138 TI - Oxcarbazepine: a review of its use in children with epilepsy. AB - Oxcarbazepine (Trileptal, Timox) is structurally related to carbamazepine and has anticonvulsant activity. Studies suggest that the anticonvulsant activity of oxcarbazepine is mediated via the blocking of neuronal ion channels. In patients aged <18 years, the efficacy of oxcarbazepine monotherapy was similar to that of phenytoin in children with partial onset or generalized tonic-clonic seizures in a 48-week trial. Additional supporting findings demonstrated that 43-71% of patients with partial onset, generalized or undetermined epilepsy were seizure free after oxcarbazepine monotherapy (mean dosage 27.7-50 mg/kg/day; duration 1-5 years). In contrast, one small nonblind trial showed more patients treated with oxcarbazepine monotherapy than with carbamazepine monotherapy had recurrent seizures during 16 months of therapy (although the conclusions that can be drawn from this trial are limited). As adjunctive therapy, oxcarbazepine was significantly better than placebo at reducing seizure frequency in children and adolescents with refractory partial onset seizures with or without secondary generalization: the median percentage change in partial onset seizure frequency was 35% vs 9%, respectively, during 16 weeks of therapy. In noncomparative trials of adjunctive oxcarbazepine (mean dosage of 34.5-56.7 mg/kg/day), 7-11% of patients with partial onset or generalized seizures were seizure free during treatment, and 20-54% had seizure reductions of > or=50%. Oxcarbazepine was generally well tolerated during monotherapy and adjunctive therapy; 2.5% and 10% of patients withdrew from well controlled trials of oxcarbazepine monotherapy and adjunctive therapy. Oxcarbazepine monotherapy was better tolerated than phenytoin and events observed in oxcarbazepine-treated patients were transient. Oxcarbazepine metabolism is largely unaffected by induction of the cytochrome (CYP) P450 system. However, oxcarbazepine can inhibit CYP2C19 and induce CYP3A4 and CYP3A5, thereby interfering with the metabolism of other drugs (e.g. phenytoin). In addition, oxcarbazepine decreases plasma levels of oral contraceptives and alternative contraceptive methods should be used. In conclusion, oxcarbazepine (as both monotherapy and adjunctive therapy) has shown efficacy in the treatment of partial onset seizures in children with epilepsy. Nevertheless, the generally favorable tolerability profile and relatively low potential for drug interactions of oxcarbazepine make it a valuable option in the treatment of childhood epilepsy. PMID- 12895140 TI - Nitric oxide donors and the skin: useful therapeutic agents? AB - Nitric oxide (NO) is involved in several processes in the skin, including wound healing, pigmentation and regulation of apoptosis. Pharmacological intervention with NO donors appears to be a promising treatment for conditions such as diabetic ulceration. In this issue of Clinical Science, Khan et al. describe the synthesis of a number of NO donors, based on nitrosylated thiosugars, structural changes in which altered the rate of NO release. The in vivo behaviour of one of the donors was then measured by laser Doppler blood flow measurements on the skin. The ability to tailor the pharmacokinetic behaviour of these drugs should be of interest in developing NO donors for clinical use. PMID- 12895143 TI - Elementary concepts of medicine. PMID- 12895141 TI - Pyridoxalated haemoglobin polyoxyethylene conjugate, a nitric oxide scavenger, decreases dose-limiting hypotension associated with interleukin-2 (IL-2) therapy. AB - Interleukin-2 (IL-2), a cytokine that induces natural killer cells termed lymphokine-activated killer (LAK) cells, is in use as an anticancer agent. During IL-2 therapy, adverse effects, such as vasodilatation and hypotension, are common. Previous studies suggest that these effects are due to nitric oxide (NO). Therefore a model of IL-2-induced hyperdynamic response in sheep was developed to test the effect of pyridoxalated haemoglobin polyoxyethylene conjugate (PHP; a NO scavenger), which is currently in clinical development for the treatment of shock associated with systemic inflammatory response syndrome. Twelve female sheep were divided into four groups (n =3 per group): sham control (Ringer's lactate solution), PHP alone (20 mg x kg(-1) x h(-1) for 96 h), IL-2 alone (recombinant human IL-2; 1,440,000 units/kg intravenously every 8 h) and a combination of PHP and IL-2. All of the sheep received Ringer's lactate solution to maintain haematocrit at baseline levels. The sheep had free access to food and water. A fall in the mean arterial pressure and systemic vascular resistance index by 20% was observed in the IL-2 group, but not in the PHP+IL-2 group. The fluid requirement to maintain the haematocrit was higher in the IL-2 group (5 ml x kg( 1) x h(-1)) than in the PHP+IL-2 group (4 ml x kg(-1) x h(-1)). The sham group showed no changes in any of the parameters. Scavenging NO by PHP prevented the hyperdynamic reaction induced by IL-2 administration in sheep. This activity of PHP may prevent the early discontinuation of IL-2 therapy that results because of these adverse events. PMID- 12895144 TI - Elementary concepts of medicine: I. Medicine: challenges with its concepts. PMID- 12895145 TI - Elementary concepts of medicine: II. Health, health fields, public health. PMID- 12895146 TI - Elementary concepts of medicine: III. Illness: somatic anomaly with... PMID- 12895147 TI - Elementary concepts of medicine: IV. Sickness from illness and in health. PMID- 12895148 TI - Elementary concepts of medicine: V. Disease: one of the main subtypes of illness. PMID- 12895149 TI - Elementary concepts of medicine: VI. Genesis of illness: pathogenesis, aetiogenesis. PMID- 12895150 TI - Elementary concepts of medicine: VII. Course of illness: manifestations, complications, outcome. PMID- 12895151 TI - Elementary concepts of medicine: VIII. Knowing about a client's health: gnosis. PMID- 12895152 TI - Elementary concepts of medicine: IX. Acting on gnosis: doctoring, intervening. PMID- 12895153 TI - Elementary concepts of medicine: X. Being a good doctor: professionalism. PMID- 12895154 TI - Elementary concepts of medicine: XI. Illness in a community: morbidity, epidemiology. PMID- 12895155 TI - Elementary concepts of medicine: XII. Specialties of medicine, genuine and other. PMID- 12895156 TI - Professionalism in medicine. AB - A Charter on Medical Professionalism (CMA) has just recently been developed internationally, and the Canadian Medical Association is calling for public dialogue on medical professionalism now that reforms in the Canadian system of health care are imminent. We posit that good practices are at issue; we outline the essence of these in general and also specifically in the knowing, teaching and intervening components of practice. We also see challenges not to, but in, medical professionalism - first and foremost in the profession's definition of good practices and the payer's confinement of insurance coverage to these. PMID- 12895158 TI - Barriers to physicians' decisions to discuss hospice: insights gained from the United States hospice model. AB - Due to its comprehensive and cost-saving design, hospice has become a critical component of health care. Physicians have become the primary gatekeepers to information on hospice and sources of referral to hospice. However, many physicians do not discuss hospice options until late in the disease course, when patients and their families are no longer able to benefit from hospice services. Although physicians, as well as patients and hospice personnel, cite patient and hospice structure factors as barriers, the present article will focus on barriers physicians have more control over, such as their discomfort discussing terminality and fear of losing contact with patients. Focusing on the American hospice model, the present article will review past findings, examine gaps in the research, and propose systematic ways to discern whether the factors physicians claim are barriers actually affect their decision making about hospice referral. PMID- 12895159 TI - Improving ACE inhibitor use in patients hospitalized with systolic heart failure: a cluster randomized controlled trial of clinical practice guideline development and use. AB - RATIONALE, AIMS AND OBJECTIVES: The efficacy of angiotensin-converting enzyme (ACE) inhibitors in treating heart failure is well established, but there is concern that these agents are underutilized. This study aimed to evaluate the effect of developing and implementing Clinical Practice Guidelines (CPGs) on the quality of care given to patients receiving ACE inhibitors for systolic heart failure. METHODS: Twenty cardiology units in Lorraine (France) were randomized to an experimental (n = 10) or a control group (n = 10). In each experimental unit, doctors were involved in drafting and implementing CPGs; those at control units were not. Practice surveys were conducted in all units before and after the intervention; 723 patients with heart failure and less than 75 years old were included. The main outcome was compliance with the CPGs. RESULTS: Before intervention, clinicians in both groups were already compliant with CPGs relating to indications and contra-indications, adverse effects management, concomitant therapy and monitoring of biologic factors. After intervention, adherence to others CPGs was generally better in the experimental group. Compliance with the CPG relating to ACE inhibitor dose on discharge was higher in the experimental group (P = 0.003). Compliance with CPGs relating to increasing ACE inhibitors doses (P < 0.0001) and the contents of the discharge letter (P = 0.02) improved in all units between the two periods. CONCLUSIONS: These results suggest that doctors involved in drafting and implementing CPGs are more likely to comply with them. PMID- 12895160 TI - In 'modest but practical ways': medical practitioners and substance misuse in Aboriginal Australians. PMID- 12895162 TI - Health effects of kava use in an eastern Arnhem Land Aboriginal community. AB - BACKGROUND: Heavy kava use in Aboriginal communities has been linked to various health effects, including anecdotes of sudden cardiac deaths. AIMS: To examine associations between kava use and potential health effects. METHODS: A cross sectional study was carried out within a kava-using east Arnhem Land Aboriginal community in tropical northern Australia. One-hundred-and-one adults who were current, recent or non-users of kava were enrolled in March 2000. Main outcome measures were physical, anthropometric, biochemical, haematological, immunological and neurocognitive assessments. RESULTS: Kava users more frequently showed a characteristic dermopathy (P<0.001). They had increased levels of gamma glutamyl transferase and alkaline phosphatase (P<0.001). Lymphocyte counts were significantly lower in kava users (P<0.001). Fibrinogen, plasminogen activator inhibitor-1 and neurocognitive tests were not different between kava use categories. IgE and IgG antibodies were elevated across the whole group, as were C-reactive protein and homocysteine. CONCLUSIONS: Kava use was associated with dermopathy, liver function abnormalities and decreased lymphocytes. If kava continues to be used by Aboriginal populations, monitoring should focus on the health consequences of these findings, including a possible increase in serious infections. The interaction between kava, alcohol and other substances requires further study. Although markers of cardiovascular risk are increased across the population, these were not higher in kava users, and this increase may be linked to the large infectious pathogen burden reflective of the socioeconomic disadvantage seen in many remote Aboriginal communities. PMID- 12895163 TI - Role of intravenous immunoglobulin in severe steroid-dependent asthma. AB - BACKGROUND: Subgroups of asthma patients have extremely severe respiratory symptoms that require chronic use of steroids for disease control. These patients are at risk of significant side-effects from chronic exposure to high doses of oral steroids. Intravenous immunoglobulin (IVIG) has immunomodulatory properties as shown by its use in some immune disorders. A few trials have suggested a possible benefit in individuals with severe asthma. AIMS: To evaluate the role of IVIG as an adjunctive therapy in steroid-dependent asthma, monitoring the outcomes of lung function and measured reduction in oral steroid requirement. METHOD: Seven patients with severe steroid-dependent asthma were given IVIG at a dose of 1 g/kg each month for 6 months. Baseline pulmonary function tests and immunoglobulin levels were obtained. At the end of 6 months, the end-points observed were lung function and the degree of reduction in the dose of oral steroids. The number of hospital admissions during the 12 months following commencement of IVIG was compared with the preceding 12 months. RESULTS: There was a significant reduction in daily prednisolone dose from 56 +/- 31 mg to 39 +/ 35 mg (P=0.04, Wilcoxon rank sum test) and a decrease in the number of hospital admissions from 5.9 +/- 2.9 to 3.6 +/- 3.5 (P=0.04). No significant improvement occurred in lung function. CONCLUSION: IVIG provides a potentially important adjunctive therapy in severe steroid-dependent asthma, reducing steroid requirement and decreasing hospital admissions, but not improving lung function. PMID- 12895164 TI - Treatment of recurrent aspiration pneumonia in end-stage dementia: preferences and choices of a group of elderly nursing home residents. AB - BACKGROUND: Clinicians are often faced with the dilemma of how best to manage patients with advanced dementia who present to the hospital with repeated episodes of aspiration pneumonia. Rarely, if at all, are the opinions of the group that is most likely to be directly affected, that is the older nursing home resident, sought. This study seeks to fill that gap. AIMS: To study the choices and preferences of a group of elderly nursing home residents in the treatment of recurrent aspiration pneumonia on a background of severe disability from end stage dementia. METHODS: A descriptive cross-sectional interview study using a hypothetical scenario carried out in six nursing homes within eastern Sydney, from June to August 2000. Views and attitudes towards hospitalization, antibiotic use, tube feeding and other treatment measures, given a situation of recurrent aspiration pneumonia and end-stage dementia, were obtained. RESULTS: Fifty-two elderly nursing home residents who were cognitively intact and not depressed were interviewed. Most of the subjects would prefer further hospital admissions (61.5%, P<0.1) and would choose to have antibiotic treatment (73.1%, P<0.001). Slightly more than half would not agree to artificial ventilation. Sixty-nine percent of the respondents would not agree to feeding via a nasogastric tube (P<0.05) and 71% would not agree to a feeding gastrostomy (P<0.001). Most would agree to a modified diet (75%, P<0.0001) and to continue oral feeding despite the attendant risk of re-aspiration (59.6%, P<0.01). CONCLUSIONS: The participants generally preferred to be treated in a hospital setting given the scenario. Most disagreed with the use of artificial feeding. PMID- 12895165 TI - Effect of different aspirin doses on platelet aggregation in patients with stable coronary artery disease. AB - BACKGROUND: Aspirin is widely used as an antiplatelet agent in the primary and secondary prevention of cardiovascular disease. In order to spare prostacyclin formation and reduce gastrointestinal side-effects, very low doses of aspirin have been introduced. However, it remains unclear whether these low doses are equally effective with respect to inhibition of platelet aggregation. AIMS: In a randomized, controlled study in 60 patients with stable coronary artery disease, the effects on platelet aggregation of five doses (50, 80, 100, 162.5 and 325 mg) of aspirin, which are widely used in clinical practice, given for 70 days, were investigated. Two reagents, adenosine diphosphate (ADP) and epinephrine, were used to induce platelet aggregation in platelet-rich plasma. An age- and sex matched group of people without coronary artery disease served as the control. RESULTS: ADP- and epinephrine-induced platelet aggregation was 78.2 +/- 12.8% and 76.7 +/- 15.5% of maximum aggregation in the control group. Aspirin inhibited platelet aggregation in a dose-dependent manner. Minimum platelet aggregation was observed at a dose of 325 mg aspirin (27.5 +/- 17.4% with ADP). Doses of 50 and 80 mg aspirin were much less effective in inhibiting platelet aggregation (59.1 +/- 11.4% and 50.3 +/- 12.1% with ADP, respectively). Doses of 100 and 162.5 mg aspirin produced significantly greater inhibition of platelet aggregation than lower doses (36.2 +/- 11.7% and 38.5 +/- 19.8% platelet aggregation with ADP, respectively). CONCLUSION: Our results demonstrate that doses of aspirin less than 100 mg are not as effective at inhibiting platelet aggregation as doses greater than 100 mg. PMID- 12895166 TI - Complication rates of colonoscopy in an Australian teaching hospital environment. AB - BACKGROUND: Quality assurance is an important part of health-care delivery. With the high level of awareness relating to adverse events from medical care, demonstration of a high standard of practice in gastroenterology is desirable. AIMS: To determine the incidence of significant complications or death within 30 days of an outpatient colonoscopy, and confirm that these are in keeping with international standards. METHODS: A retrospective audit of linked endoscopy and other hospital databases and selected medical records was carried out, based on reports of 30,463 colonoscopies performed between 5 September 1989 and 31 December 1999 in the three Western Australian public teaching hospitals. RESULTS: A total of 23,508 colonoscopies was performed on an outpatient basis between 5 September 1989 and 31 December 1999. Post-procedural complications identified (and incidence) were: bleeding episodes 49 (0.21%), colonic perforation 23 (0.1%), abdominal pain 22 (0.09%), and others 19 (0.08%). A total of 196 patients died within 30 days of undergoing colonoscopy (0.83%), although only three deaths were attributable to the procedure itself (incidence 0.01%). Two were inpatients at the time of the procedure (outpatient mortality rate 0.004%). The combined incidence of bleeding and perforation was not significantly different between consultant endoscopists and unassisted trainees (incidence 0.21% vs 0.20%, P=0.98). CONCLUSIONS: The incidence of bleeding and perforation is similar to other reported series and reflects procedures performed by personnel with a wide range of endoscopic experience. The incidence of complications was not greater for trainees compared with consultant endoscopists. All bleeding episodes and the majority of perforations were associated with a therapeutic intervention. Diagnostic colonoscopy in particular is a very safe procedure. PMID- 12895167 TI - Future directions of medical education in Australia. PMID- 12895168 TI - Ethical aspects of blood and organ donation. AB - Ethics of blood and organ donation in Australia has been influenced by voluntarism and the Red Cross. The General Agreement on Trade in Services may threaten the principle of voluntary blood donation and self sufficiency of blood in Australia. However, a cooperative approach to managing the public good is more sustainable than individual self interest and the author argues that voluntary blood and organ donation should be retained. PMID- 12895169 TI - An update on asthma management. AB - Asthma remains a major cause of morbidity in the Australian community, despite enhanced strategies and interventions for achieving optimal outcomes. Although there is some evidence of over use of high doses of inhaled corticosteroids (ICS), there is also evidence for under use of ICS, despite long-term data demonstrating clear benefits of low doses. The present paper briefly discusses some of the issues that are pertinent to the development of sensitization and atopic disease, but focuses mainly on the current understanding of best clinical practice for adults with asthma and the optimal approach to management. A clear definition of asthma control and a systematic approach to down-titration to minimize ICS doses is crucial to achieving better outcomes. Patient self management education and optimal therapy are the keys to achieving better outcomes, although uncertainty remains about recommendations for mild asthma, despite new data. PMID- 12895170 TI - Counting the costs of treatment: the reproductive and gynaecological consequences of adjuvant therapy in young women with breast cancer. AB - As the mortality rate from breast cancer decreases, the issues facing breast cancer survivors are becoming increasingly important. Survivors of all ages may face physical and psychosocial consequences of their diagnosis and treatments. However, the long-term fertility and menopause-related side-effects of adjuvant therapy uniquely affect younger premenopausal breast cancer survivors. This article provides an evidence-based overview of the reproductive and gynaecological impact of breast cancer therapy for premenopausal women diagnosed with breast cancer. The physical and psychosocial implications of premature menopause are presented. Strategies for preserving fertility in selected patients are also discussed. Recent clinical trials strongly indicate that premenopausal women with oestrogen receptor positive tumours should receive endocrine therapy. The increased use of endocrine therapies in younger women raises important questions regarding patient information needs and treatment decision-making. PMID- 12895171 TI - Cellular adhesion molecules and cardiovascular disease. Part I. Their expression and role in atherogenesis. AB - Atherosclerosis is now recognized to be an inflammatory disease, and several inflammatory markers have been shown to be associated with both the presence and future risk of symptomatic cardiovascular disease. Cellular adhesion molecules, particularly members of the selectin family and immunoglobulin superfamily, are intimately involved in the recruitment of leucocytes to sites of inflammation, including developing atherosclerotic lesions. Their role in the pathogenesis of atherosclerosis has been clearly demonstrated using knockout mice models. Plasma levels of adhesion molecules, which have been shed from the cell surface, have been associated with the presence of clinical atherosclerotic disease, although published studies differ in their findings. This limited consensus in the literature may be explained either by unrecognized confounding factors, or perhaps by the unpredictable relationship between cell surface expression and activity of cellular adhesion molecules and their shedding into the plasma. While cell surface activity of adhesion molecules appears critical in the development of atherosclerotic lesions, the measurement of plasma levels of soluble adhesion molecules may offer little additional benefit for individual patients in the prediction of the extent of atherosclerotic disease above the assessment of conventional cardiovascular risk factors. PMID- 12895172 TI - An unusual presentation of synovial cyst of the right knee. PMID- 12895173 TI - Medication use among older Australian veterans and war widows. AB - This study aimed to review the medications used by a group of Australian war veterans and widows, to identify type and number of medications used and potential inappropriate prescribing. Label details of 4304 medications used by 887 participants were recorded and classified according to the Anatomical Therapeutic Chemical code. The median number of regular medications being used was four (interquartile range 2-7), with a median of one other medication (interquartile range 1-2) being taken as needed. Over half (53%) the participants were taking four or more regular medications. There were 26 potential interactions among 25 people. Polypharmacy was a considerable problem for this group of older Australians. PMID- 12895174 TI - Health and stress in Australian interns. AB - Over a 3-year period, interns completed questionnaires on self-reported health and stress during their orientation week at the beginning of the year, during mid year, and at the end of the first year. Interns experienced increased levels of stress and ill health in the middle of the year as compared with the start and end of the year (chi2=11.9, P=0.003 and chi2=20.4, P<0.001, respectively). Despite workplace reform for junior doctors, this study suggests that interns still feel overburdened with work expectations and this is having a negative effect on their health. PMID- 12895175 TI - Giant cell arteritis presenting with scalp necrosis. PMID- 12895176 TI - Cicatricial pemphigoid in non-Hodgkin's lymphoma. PMID- 12895177 TI - Helicobacter pylori hiccup. PMID- 12895179 TI - Helicobacter pylori hiccup. PMID- 12895180 TI - Ciclopirox gel: an update. PMID- 12895181 TI - Ciclopirox for the treatment of superficial fungal infections: a review. AB - Ciclopirox is a broad-spectrum antifungal agent that also exhibits anti inflammatory and antibacterial activity. The lotion and cream formulations of ciclopirox are effective in many types of infection, including tinea corporis/cruris, tinea pedis, cutaneous candidiasis, pityriasis (tinea) versicolor, and seborrheic dermatitis. The new ciclopirox gel 0.77% formulation is also indicated for the treatment of seborrheic dermatitis of the scalp, interdigital tinea pedis and tinea corporis. PMID- 12895182 TI - Evaluation of in vitro activity of ciclopirox olamine, butenafine HCl and econazole nitrate against dermatophytes, yeasts and bacteria. AB - BACKGROUND: In many instances, a cutaneous fungal infection may exist concomitantly with bacterial involvement. In this study we compared the in vitro activity of three antifungal agents against the dermatophytes, yeasts and bacteria recovered most commonly from cutaneous mycoses and bacterial infections. METHODS: Using a microdilution method adapted from the National Committee for Clinical Laboratory Standards (NCCLS), we determined the minimum inhibitory concentrations (MICs) of ciclopirox olamine, econazole nitrate and butenafine HCl against a panel of dermatophyte fungi and yeasts (n = 39) and bacterial isolates (n = 45). RESULTS: All three antifungals demonstrated comparable activity against the dermatophytes tested, with a MIC range of 0.03-0.25 micro g/ml for ciclopirox, < 0.001-0.25 micro g/ml for econazole and 0.03-0.25 micro g/ml for butenafine. For yeasts, ciclopirox showed activity against all isolates, with an MIC range of 0.001-0.25 micro g/ml, whereas econazole had a broader range of 0.125-> 0.5 micro g/ml. Butenafine displayed limited activity against the yeast Candida albicans and no activity against Malassezia furfur. For the antibacterial activity studies, ciclopirox demonstrated activity against all isolates tested with a range of 0.06-2 micro g/ml, while econazole showed activity against Gram positive bacteria only, with a MIC range of 0.004-0.25 micro g/ml. Butenafine HCl had a limited activity against bacterial isolates tested, showing activity against beta-hemolytic Streptococcus Group A and Corynebacterium only. Neither econazole nitrate nor butenafine HCl demonstrated activity against any of the Gram-negative strains evaluated in this study. CONCLUSIONS: The data suggest that ciclopirox olamine has the broadest in vitro activity, in comparison to econazole and butenafine HCl, against bacteria, yeasts and bacteria. These findings may have implications in the use of these antimycotics in the treatment of mixed cutaneous infections where bacteria or yeasts are present in addition to dermatophytes. PMID- 12895183 TI - Ciclopirox gel for seborrheic dermatitis of the scalp. AB - BACKGROUND: Seborrheic dermatitis is a common inflammatory skin disorder that usually occurs in patients with pre-existing seborrhea. The etiology of seborrheic dermatitis is uncertain. Typically, sites dense with sebaceous glands support growth of the lipophilic yeast Malassezia furfur. Ciclopirox (Loprox) gel is a hydroxypyridone, broad-spectrum antifungal agent proven effective against the yeast M. furfur. OBJECTIVE: A multicenter, randomized, double-blind, vehicle controlled study of 178 subjects evaluated the efficacy of ciclopirox gel in treating seborrheic dermatitis of the scalp. METHODS: One hundred and seventy eight subjects were randomized to apply either ciclopirox gel 0.77% twice daily, or vehicle twice daily for 28 days. Subjects' signs and symptoms of severity (erythema, scaling, pruritus and burning) were rated on a scale of 0-3 (none to severe); for inclusion, a minimum score of 4, for the sum of the individual ratings was required. Efficacy evaluations were performed at baseline, days 4, 8, 15, 22, 29, and at end-point (final visit, up to day 33). The primary efficacy variable was clinical response assessed by a global improvement, based on a scale of 0-5 (100% clearance to flare of treatment area). Changes in signs/symptoms severity scores within the target lesion were also evaluated. RESULTS: Global evaluation scores demonstrated that significantly more ciclopirox-treated subjects achieved over 75% improvement compared with vehicle at days 22, 29, and endpoint (P < 0.01). Change-from-baseline mean score for total signs and symptoms was significantly greater in ciclopirox subjects compared with vehicle subjects at the same time points as above (P < 0.001), as well as day 15 (P < 0.01). Twenty-nine percent of subjects rated ciclopirox as having excellent cosmetic acceptability. There were only mild adverse events, with the most common being burning sensation in 13% of ciclopirox subjects and 9% of vehicle subjects. CONCLUSION: Ciclopirox gel is effective and safe in the treatment of seborrheic dermatitis of the scalp. PMID- 12895184 TI - Interdigital tinea pedis (dermatophytosis simplex and complex) and treatment with ciclopirox 0.77% gel. AB - The most common presentation of tinea pedis (athlete's foot) is that involving the interdigital spaces. Tinea pedis interdigitalis may present as asymptomatic dermatophytosis simplex or dermatophytosis complex, which is symptomatic, with secondary bacterial infection. In the dermatophytosis complex presentation there may be inflammation, maceration and odor, with bacterial involvement. Ciclopirox gel offers advantages in the treatment of tinea pedis, especially in the dermatophytosis complex presentation, with antifungal, antibacterial, and anti inflammatory activity; furthermore, the gel formulation is fast drying, which is an advantage when the toe web area is moist. PMID- 12895186 TI - British Journal of Clinical Pharmacology review series. Clinical research methods in gastroenterology. PMID- 12895187 TI - Assessment of non-steroidal anti-inflammatory drug (NSAID) damage in the human gastrointestinal tract. AB - Aspirin is widely prescribed and confers considerable benefit to patients by reducing cardiovascular and cerebrovascular morbidity and mortality. Nonsteroidal anti-inflammatory drugs (NSAIDs) are effective analgesics, antipyretics and reduce the inflammatory component in conditions such as rheumatoid arthritis. However, both agents are associated with an increased risk of gastrointestinal symptoms and the potentially serious consequences of gastroduodenal ulceration, bleeding and perforation. The introduction of highly selective cyclo-oxygenase (COX)-2 inhibitors or the coprescription gastroprotective agents with nonselective NSAIDs have offered strategies to reduce the incidence of such events. This review article analyzes the quantitative techniques that can be employed by clinical pharmacologists and the clinical studies performed to assess NSAID damage in the gastrointestinal tract. PMID- 12895188 TI - Gastric function measurements in drug development. AB - The function of the stomach includes initiation of digestion by exocrine secretions such as acid and pepsin, which are under the control of the endocrine secretion of hormones that also coordinate intestinal motility. The stomach also stores and mechanically disrupts ingested food. Various techniques have been developed to assess gastric physiology, the most important of which is assessment of acid secretion, as well as gastric motility and gastric emptying. The influence of drugs on gastric function and the effect of gastric secretion and mechanical actions on the bioavailability of novel compounds are of critical importance in drug development and hence to clinical pharmacologists. The control of acid secretion is essential in the treatment of peptic ulcer disease as well as gastrooesophageal reflux disease (GORD); pH-metry can be used to determine the necessary dose of an acid suppressant to heal mucosal damage. Disturbed gastric myoelectric activity leading to gastroparesis can cause delayed gastric emptying, often found in patients with diabetes mellitus. Electrogastrography (EGG) may be used to evaluate the influence of prokinetics and other drugs on this condition and aid in determining effective therapy. PMID- 12895189 TI - A simple pain model for the evaluation of analgesic effects of NSAIDs in healthy subjects. AB - AIMS: Non-steroidal anti-inflammatory drugs (NSAIDs) are believed to counteract inflammation and inflammation-induced sensitization of nociceptors by inhibiting peripheral prostaglandin synthesis. We evaluated an experimental pain model for NSAIDs, that included an inflammatory component to mimic clinical inflammatory pain conditions. METHODS: The study was performed in a randomized, double-blind, placebo-controlled, two-way crossover design on 32 healthy volunteers. A small skin area of the proximal upper leg was irradiated with a UVB source using three times the individually estimated minimal erythema dose. Twenty hours after irradiation skin temperature, heat pain threshold and tolerance in sunburn spot were measured using a thermal sensory testing. These measurements were repeated 2 h after medication of either 800 mg ibuprofen as single oral dose or placebo capsules. Effects of ibuprofen on outcome parameters were assessed with analyses of covariance (ancova). RESULTS: Placebo did not affect heat pain threshold or tolerance. By contrast, ibuprofen increased heat pain threshold by 1.092 degrees C [confidence interval (CI) 0.498, 1.695; P = 0.0008) compared with placebo. Heat pain tolerance also increased significantly by 1.618 degrees C (CI 1.062, 2.175; P = 0.0001). CONCLUSION: The pain model we evaluated was well tolerated in all subjects and the effects of ibuprofen were highly significant. This model is simple, sensitive to NSAIDs' effects and therefore has potential for future experimental pain studies. PMID- 12895190 TI - Population pharmacokinetics of clomethiazole and its effect on the natural course of sedation in acute stroke patients. AB - AIMS: This analysis was performed to investigate the population pharmacokinetics of clomethiazole and its effect on the natural course of sedation in acute stroke patients using a nonlinear mixed effects modelling approach. METHODS: One thousand five hundred and forty-six acute stroke patients (774 on active treatment) from 166 centres were included in three randomized, double-blind, placebo-controlled phase III efficacy and safety studies. A total dose of 68 mg kg(-1) clomethiazole edisilate was given as a three-phase i.v.-infusion over 24 h. Three blood samples were drawn from all patients to characterize the pharmacokinetics. Sedation was monitored throughout the entire treatment period and the degree of sedation was measured on a discrete ordinal scale with six levels. Models were fitted to the data using the software NONMEM. RESULTS: Clomethiazole was characterized by a two-compartment pharmacokinetic model with interindividual variability in all structural parameters. For a patient weighing 75 kg, the average CL, V1, Q, and V2 was estimated to be 52.7 l h(-1), 82.5 l, 167 l h(-1) and 335 l, respectively. The interindividual variability in CL, V1, Q and V2 was estimated to be 48%, 53%, 42% and 54%, respectively. Increasing body weight and concomitant administration of liver enzyme inducing drugs were found to increase clearance (by 0.5 l h(-1) kg(-1) and 40%, respectively). Increasing weight also increased the volume of distribution (1.1 l kg(-1) for V1 and 4.7 l kg(-1) for V2). A six-category proportional odds model with a component including the natural course of sedation following placebo administration, a drug component (present or absent) and an interindividual variability component described the degree of sedation. Stroke severity as measured on the NIH-stroke scale on admission and drug treatment were the most important predictors of sedation, but a nonlinear increase in sedation with increasing age was also found. Increasing body weight increased the sedative drug effect. CONCLUSIONS: The pharmacokinetics of clomethiazole were characterized in acute stroke patients and the analysis excluded several possible covariates of interest in drug development. The time course of sedation could be quantitatively described during the first 24 h following an acute stroke in the presence or absence of clomethiazole treatment. PMID- 12895191 TI - Olanzapine pharmacokinetics are similar in Chinese and Caucasian subjects. AB - AIM: To compare the pharmacokinetic profiles and dose proportionality of olanzapine in Chinese and Caucasian subjects. METHODS: Randomized, three-period study with 12 Chinese and 12 Caucasian, healthy, male subjects administered 2.5, 5 and 10 mg olanzapine. Noncompartmental pharmacokinetic parameters were derived. RESULTS: No statistically significant racial differences in the weight-normalized pharmacokinetic parameters were observed except for Vz/Fnorm, which was 17% lower at the 5- and 10-mg dose in the Chinese group (95% confidence interval 8.49, 10.1 and 8.05, 9.73, respectively), compared with the Caucasian group (9.53, 12.8 and 9.39, 12.0, respectively). Olanzapine's pharmacokinetics were linear and dose proportional in both racial groups. CONCLUSION: The pharmacokinetics of olanzapine are similar in both Chinese and Caucasian racial groups. PMID- 12895192 TI - Population pharmacokinetic analysis of mycophenolic acid in renal transplant recipients following oral administration of mycophenolate mofetil. AB - AIM: To develop a population pharmacokinetic model for mycophenolic acid in adult kidney transplant recipients, quantifying average population pharmacokinetic parameter values, and between- and within-subject variability and to evaluate the influence of covariates on the pharmacokinetic variability. METHODS: Pharmacokinetic data for mycophenolic acid and covariate information were previously available from 22 patients who underwent kidney transplantation at the Princess Alexandra Hospital. All patients received mycophenolate mofetil 1 g orally twice daily. A total of 557 concentration-time points were available. Data were analysed using the first-order method in NONMEM (version 5 level 1.1) using the G77 FORTRAN compiler. RESULTS: The best base model was a two-compartment model with a lag time (apparent oral clearance was 27 l h(-1), and apparent volume of the central compartment 98 l). There was visual evidence of complex absorption and time-dependent clearance processes, but they could not be successfully modelled in this study. Weight was investigated as a covariate, but no significant relationship was determined. CONCLUSIONS: The complexity in determining the pharmacokinetics of mycophenolic acid is currently underestimated. More complex pharmacokinetic models, though not supported by the limited data collected for this study, may prove useful in the future. The large between-subject and between-occasion variability and the possibility of nonlinear processes associated with the pharmacokinetics of mycophenolic acid raise questions about the value of the use of therapeutic monitoring and limited sampling strategies. PMID- 12895193 TI - Effects of sex on the pharmacokinetic and pharmacodynamic properties of quinidine. AB - AIMS: To investigate the source of the apparent increased susceptibility of women to develop QT interval prolongation and torsade de pointes after the administration of drugs that delay cardiac repolarization. METHODS: Plasma quinidine concentrations and electrocardiographic changes (QRS and QT intervals) were measured over 24 h following the administration of single oral doses of the QT prolonging drug quinidine (3 mg kg(-1)) and compared between 27 male and 21 female healthy volunteers. RESULTS: There were no significant differences between males and females in plasma quinidine concentrations or in calculated pharmacokinetic variables. Maximum quinidine concentrations in males and females were 997 +/- 56 and 871 +/- 57 ng ml(-1), respectively (mean difference (-125, 95% confidence intervals (CI) -239, 11 ng ml(-1), P = NS). Quinidine lengthened actual (QTa) and corrected (QTc) QT intervals and the QRS interval to a greater extent in females than males (P < 0.001 for each), but there were no significant sex differences detected in the effects of quinidine on the heart rate corrected JT interval. Maximum prolongation of QTc interval was observed 2 h after quinidine and was significantly greater in women (33 +/- 16 vs 24 +/- 17 ms, mean difference 9 +/- 20 ms, 95% CI 3, 15, P = 0.037). At this time mean differences (95% CI) were 1.0 min(-1) (-2.5, 4.4, P = NS) for heart rate, 5.5 ms (3.5, 7.6, P = 0.05) for the QRS and 3.4 ms (-2.5, 9.3, P = NS) for the JTc intervals. CONCLUSIONS: Quinidine-induced increases in QTc were larger in females, but no sex differences in quinidine pharmacokinetics were found. The disparity in prolongation of cardiac repolarization is thus due to a pharmacodynamic difference which appears more complex than simply an increase in repolarization delay in females. PMID- 12895194 TI - Population pharmacokinetics of platinum after nedaplatin administration and model validation in adult patients. AB - AIMS: The pharmacokinetics of unbound platinum after administration of an anticancer drug nedaplatin, cis-diammineglycolateplatinum were examined using population analysis. The relevant covariates and the extent of inter- and intra individual variability were evaluated. METHODS: In order to clarify the pharmacokinetic profile of nedaplatin, unbound platinum concentrations (789 points) in plasma after intravenous infusion of nedaplatin were obtained from 183 courses for 141 patients. Plasma concentration data were analysed by nonlinear mixed effect modelling using NONMEM to evaluate the population mean parameters and variances for inter- and intra-individual random effects. The final population model was validated by parameter sensitivity analysis using objective function mapping, the bootstrap resampling and a data-splitting technique, i.e. the Jackknife method, and the predictive performance of the final model was evaluated. RESULTS: A two-compartment pharmacokinetic model with zero-order input and first order elimination described the current data well. The significant covariates were creatinine clearance (CLcr) for clearance of platinum (CL) [population mean [95% confidence interval (CI)] CL (l h(-1)) = 4.47 (3.27, 5.67) + 0.0738 (0.0581, 0.0896) x CLcr (CLcr: ml min(-1))] and body weight (BW: kg) for volume of distribution of platinum (Vc) [Vc (l) = 12.0 (7.5, 16.5) + 0.163 (0.081, 0.246) x BW]. Inter-individual variations (CV%, 95% CI) for CL and Vc were 25.5% (20.7, 29.6) and 21.4% (17.0, 24.1), respectively, and intra individual variation (CV%, 95% CI) was 12.6% (10.5, 14.4). The effects of pretreatment with nedaplatin or other platinum agents on clearance and volume of distribution were also tested, but no significant effect was found. The relationship between the observed and predicted unbound platinum concentration by empirical Bayesian prediction showed good correlation with no bias, suggesting that the final model explains well the observed data in the patients. The mean prediction error and root mean square prediction error (95% CI) were - 0.0164 micro g ml(-1) (- 0.4379, 0.4051) and 0.2155 micro g ml(-1) (not calculable, 0.6523), respectively. The values of mean, standard error and 95% CI for objective function mapping, the bootstrap resampling, the Jackknife estimates and the final model coincided well. CONCLUSIONS: A population pharmacokinetic model was developed for unbound platinum after intravenous infusion of nedaplatin. Only creatinine clearance was found to be a significant covariate of clearance, and BW was found to be a significant covariate of volume of distribution. These population pharmacokinetic estimates are useful for setting initial dosing of nedaplatin using its population mean and can also be used for setting appropriate dosage regimens using empirical Bayesian forecasting. PMID- 12895195 TI - Tacrolimus and cerivastatin pharmacokinetics and adverse effects after single and multiple dosing with cerivastatin in renal transplant recipients. AB - AIMS: In contrast to cyclosporin, only limited information exists on the interaction potential between the immunosuppressive agent tacrolimus and HMG-CoA reductase inhibitors, which are metabolized via the cytochrome P450 system. The aim of this study was to investigate the pharmacokinetics, and adverse effects of cerivastatin combined with tacrolimus in renal transplant patients. METHODS: Ten patients with stable kidney graft functions and LDL-cholesterol serum concentrations > 110 mg dl-1 were included in the study. After an observation period of 3 months, cerivastatin (0.2 mg daily) was administered for an additional 3 months. Tacrolimus steady-state pharmacokinetics and cerivastatin single- and multiple-dose pharmacokinetics were determined. Lipid concentrations, routine laboratory parameters and adverse events were obtained and analysed throughout the study period of 6 months. RESULTS: Blood tacrolimus trough concentrations were not affected by cerivastatin (mean +/- SD 8.6 +/- 2.1 ng ml( 1) before, and 8.7 +/- 2.4 ng ml(-1) at day 90 of cerivastatin dosing, with a 95% confidence interval on the difference = 0.97, 1.08). The mean area under the blood concentration-time curve to 24 h (AUC(0,24 h)) for cerivastatin was 14.5 +/ 2.53 micro g l(-1) h(-1) at day 1 after starting treatment and 19.02 +/- 3.55 micro g l(-1) h(-1) (3 months later), resulting in a 35% higher (AUC(0,24 h)) compared with the first dose. Total cholesterol, LDL-cholesterol and triglyceride concentrations were significantly lowered by cerivastatin whereas no significant effect of cerivastatin on serum creatininkinase concentrations was observed and no adverse effects were documented. CONCLUSIONS: Tacrolimus increased the AUC(0, 24 h) of cerivastatin by a mean of 35% in renal transplant patients. Cerivastatin had no detectable effect on the pharmacokinetics of tacrolimus. PMID- 12895196 TI - A discordance between cytochrome P450 2D6 genotype and phenotype in patients undergoing methadone maintenance treatment. AB - AIMS: To assess CYP2D6 activity and genotype in a group of patients undergoing methadone maintenance treatment (MMT). METHODS: Blood samples from 34 MMT patients were genotyped by a polymerase chain reaction-based method, and results were compared with CYP2D6 phenotype (n = 28), as measured by the molar metabolic ratio (MR) of dextromethorphan (DEX)/dextrorphan (DOR) in plasma. RESULTS: Whereas 9% of patients (3/34) were poor metabolizers (PM) by genotype, 57% (16/28) were PM by phenotype (P < 0.005). Eight patients, who were genotypically extensive metabolizers (EM), were assigned as PM by their phenotype. The number of CYP2D6*4 alleles and sex were significant determinants of CYP2D6 activity in MMT patients, whereas other covariates (methadone dose, age, weight) did not contribute to variation in CYP2D6 activity. CONCLUSIONS: There was a discordance between genotype and in vivo CYP2D6 activity in MMT patients. This finding is consistent with inhibition of CYP2D6 activity by methadone and may have implications for the safety and efficacy of other CYP2D6 substrates taken by MMT patients. PMID- 12895197 TI - Dose-response relationship and reproducibility of urinary salbutamol excretion during the first 30 min after an inhalation. AB - AIMS: To determine the reproducibility and dose-response relationship for urinary salbutamol excretion post inhalation. METHODS: Fifteen volunteers inhaled either one, two, three, four or five doses of 100 micro g salbutamol on separate days and then seven of these also repeated each of the one, three and five doses on five occasions. After each study dose urine was collected 30 min post inhalation. RESULTS: The mean (SD) 30 min urinary salbutamol after one, two, three, four and five doses was 2.61 (1.0.), 5.47 (1.59), 8.68 (2.73), 12.34 (3.96) and 15.99 (4.50) micro g, respectively. Statistical analysis revealed a linear relationship (P < 0.001). Mean (SD) coefficient of variation after one, three and five doses was 10.5 (3.6), 10.1 (2.7) and 9.4 (2.3)%. CONCLUSIONS: The 30 min salbutamol urinary excretion post inhalation pharmacokinetic method, to compare inhaled products, is linear with inhaled dose and reproducible. PMID- 12895198 TI - Pharmacokinetics of morphine are not altered in subjects with Gilbert's syndrome. AB - AIMS: To verify that Gilbert's syndrome, which is caused by decreased glucuronidation capacity of the UDP-glucuronosyl transferase (UGT)1A1, does not account for impaired morphine clearance. METHODS: Noncompartmental pharmacokinetic parameters for morphine and its glucuronide metabolites were compared between five carriers of Gilbert's syndrome and six noncarriers after a 7.5 mg (19.8 micro mol) intravenous injection of morphine sulphate pentahydrate. To estimate the amount of morphine-6-glucuronide (M6G) formed from morphine, 1 mg of deuterized M6G was injected intravenously at the same time. RESULTS: No differences were detected between carriers and noncarriers of Gilbert's syndrome in the clearance of morphine (80.1 +/- 12 l h(-1) vs 87.9 +/- 22 l h(-1)) and in the percentage of morphine that was metabolized to M6G (10.9 +/- 1.4 vs 13 +/- 2). The areas under the plasma concentration vs time curves of morphine, M6G and morphine-3-glucuronide also did not differ between carriers and noncarriers of Gilbert's syndrome. CONCLUSIONS: Gilbert's syndrome is not a factor to be considered when prescribing morphine. PMID- 12895199 TI - The effect of ethinyloestradiol and levonorgestrel on the CYP2C19-mediated metabolism of omeprazole in healthy female subjects. AB - AIMS: To study the effect of an oral contraceptive (OC) formulation containing ethinyloestradiol and levonorgestrel (LNG) (combination OC) or LNG alone on the CYP2C19-mediated hydroxylation of omeprazole in healthy females. METHODS: This was an open crossover study with three phases. In phase one, 10 healthy females received a single 40-mg dose of omeprazole. Thereafter the subjects received in a random order either 40 micro g ethinyloestradiol and 75 micro g LNG or 60 micro g LNG alone once daily for 10 days. On day 10, 1 h after the last OC dose, subjects received a single 40-mg oral dose of omeprazole. The plasma concentrations of omeprazole, 5'-hydroxyomeprazole and omeprazole sulphone were determined for up to 8 h. RESULTS: The use of combination OC increased the area under the curve (AUC) of omeprazole by 38% [95% confidence interval (CI) - 3.8, 80; P = 0.040] and caused a 48% increase (95% CI 28, 68) in the AUC ratio of omeprazole/5 hydroxyomeprazole. LNG alone did not effect the 5'-hydroxylation of omeprazole. Neither of the OC preparations seemed to have an inhibitory effect on the formation of omeprazole sulphone by CYP3A4. CONCLUSIONS: Oral contraceptives containing ethinyloestradiol but not those containing only LNG decrease CYP2C19 activity. PMID- 12895200 TI - A case of autoimmune polyglandular syndrome developed after interferon-alpha therapy. PMID- 12895201 TI - Risk factors in relation to an emergence of bipolar disorder: a systematic review. AB - OBJECTIVE: There is a consensus that genetic factors are important in the causation of bipolar disorder (BPD); however, little is known about other risk factors in the aetiology of BPD. Our aim was to review the literature on such risk factors - risk factors other than family history of affective disorders - as predictors for the initial onset of BPD. METHODS: We conducted a literature search using the MEDLINE, PsycINFO and EMBASE databases. We selected factors of interest including demographic factors, factors related to birth, personal, social and family backgrounds, and history of medical conditions. The relevant studies were extracted systematically according to a search protocol. RESULTS: We identified approximately 100 studies that addressed the associations between antecedent environmental factors and a later risk for BPD. Suggestive findings have been provided regarding pregnancy and obstetric complications, winter-spring birth, stressful life events, traumatic brain injuries and multiple sclerosis. However, evidence is still inconclusive. Childbirth is likely to be a risk factor. The inconsistency across studies and methodological issues inherent in the study designs are also discussed. CONCLUSION: Owing to a paucity of studies and methodological issues, risk factors of BPD other than family history of affective disorders have generally been neither confirmed nor excluded. We call for further research. PMID- 12895202 TI - A prepubertal and early adolescent bipolar disorder-I phenotype: review of phenomenology and longitudinal course. AB - OBJECTIVE: Phenomenology, assessment, longitudinal, and psychosocial findings from an ongoing, controlled, prospective study of 93 subjects with a prepubertal and early adolescent bipolar disorder phenotype (PEA-BP) will be reviewed. METHODS: Unlike adult-onset bipolar disorder, for which there were over 50 years of systematic investigations, there were a paucity of rigorous data and much controversy and skepticism about the existence and characteristics of prepubertal onset mania. With this background, issues to address for investigation of child onset mania included the following: (i) What to do about the differentiation of mania from attention-deficit hyperactivity disorder (ADHD). (ii) How to deal with the ubiquity of irritability as a presenting symptom in multiple child psychiatry disorders. (iii) Development of a research instrument to assess prepubertal manifestations of adult mania (i.e. children do not 'max out' credit cards or have four marriages). (iv) How to distinguish normal childhood happiness and expansiveness from pathologically impairing elated mood and grandiosity. RESULTS: To address these issues, a PEA-BP phenotype was defined as DSM-IV mania with elated mood and/or grandiosity as one inclusion criterion. This criterion ensured that the diagnosis of mania was not made using only criteria that overlapped with those for ADHD, and that subjects had at least one of the two cardinal symptoms of mania (i.e. elated mood and grandiose behaviors). Subjects were aged 10.9 years (SD = 2.6) and age of onset of the current episode at baseline was 7.3 years (SD = 3.5). Validation of PEA-BP was shown by reliable assessment, 6-month stability, and 1- and 2-year diagnostic longitudinal outcome. PEA-BP resembled the severest form of adult-onset mania by presenting with a chronic, mixed mania, psychotic, continuously (ultradian) cycling picture. CONCLUSION: Counterintuitively, typical 7-year-old children with PEA-BP were more severely ill than typical 27 year olds with adult-onset mania. Moreover, longitudinal data strongly supported differentiation of PEA-BP from ADHD. PMID- 12895203 TI - Decreased hippocampal NMDA, but not kainate or AMPA receptors in bipolar disorder. AB - OBJECTIVES: The purpose of this study was to determine whether there are changes in the density of ionotropic glutamate receptors in the hippocampus of subjects with bipolar disorder. METHODS: Using in situ radioligand binding with semiquantitative autoradiography, we measured the density of [3H]MK-801, [3H]CGP39653, [3H]AMPA and [3H]kainate binding in hippocampi, obtained postmortem, from eight subjects with type 1 bipolar disorder and 8 age- and sex matched controls. RESULTS: In subjects with bipolar disorder there were significant decreases in the density of [3H]MK-801 binding in the Cornu Ammonis (CA) 3 (mean +/- SEM; 108.8 +/- 12.2 versus 166.2 +/- 18.0 fmol/mg ETE, p < 0.005) as well as the pyramidal (102.8 +/- 9.2 versus 136.6 +/- 11.2 fmol/mg ETE, p < 0.05) and polymorphic (21.73 +/- 6.5 versus 53.26 +/- 11.6 fmol/mg ETE, p < 0.05) layers of the subiculum. In addition, two-way analysis of variance (ANOVA) revealed a decrease in the density of [3H]CGP39653 binding across the hippocampal formation in bipolar subjects, which did not reach significance in any subregion. There were no changes in the densities of [3H]AMPA or [3H]kainate binding in these subjects. CONCLUSIONS: [3H]CGP39653 and [3H]MK-801 bind to the glutamate binding site and open ion channel of the n-methyl-d-aspartate (NMDA) receptor, respectively. Therefore, these data suggest that there is a decrease in the number of open ion channels associated with no significant change in the apparent density of NMDA receptors in regions of the hippocampus from subjects with bipolar disorder. PMID- 12895204 TI - GAMIAN-Europe/BEAM survey I--global analysis of a patient questionnaire circulated to 3450 members of 12 European advocacy groups operating in the field of mood disorders. AB - OBJECTIVES: GAMIAN-Europe is a pan-European federation of national patient organizations from 30 European countries covering the whole spectrum of psychiatry. To gain a better understanding of what it is like to live with bipolar disorder (BD), GAMIAN-Europe undertook a detailed patient-based questionnaire, known as 'GAMIAN-Europe/BEAM Survey', examining a variety of aspects. METHODS: The questionnaire was mailed to 3450 patients from 12 member organizations in 11 countries. A total of 1760 completed questionnaires were received but 28 were ruled out as inappropriate. Of the remainder, 1041 respondents stated that they had been, or were, suffering from BD. The remainder stated that they were suffering from depression, dysthymia or atypical depression. RESULTS: The findings indicate that, on average, a bipolar patient is expected to wait for 5.7 years for a correct diagnosis from the first onset of symptoms. Many patients have a family history of mood and anxiety disorders. They experience a high degree of stigmatization from all quarters. This is reflected in the difficulties they experience in obtaining employment despite high academic achievement. Most patients receive combination therapy. Compliance problems resulting from adverse side-effects are less significant than in the past. Overall, the level of satisfaction with pharmacotherapy was high yet, paradoxically, patients had reservations about dependency issues and possible long-term side-effects. CONCLUSIONS: There was a clear need for more patient education about pharmacological and psychosocial interventions, despite material progress having been made over the past decade. There is an urgent need for more information and education for both relatives and the public in most European countries to improve awareness and understanding of BDs and other mood disorders and the doctor-patient dialogue. PMID- 12895205 TI - Approximate entropy of symptoms of mood: an effective technique to quantify regularity of mood. AB - OBJECTIVES: Several psychiatric conditions are associated with lability of affect. In this study, we investigated regularity of mood using APEN (Approximate Entropy) on daily subjective ratings using a Visual Analog Scale with 11 items pertaining to mood. METHODS: APEN was applied to the data in a double-blind placebo controlled investigation on the effects of fluoxetine (n=19), pemoline (n=18) and placebo (n=20) in normal controls. These subjects rated their subjective feelings daily at the end of each day. We analysed 56 data point time series (each value was obtained on each day) after the three experimental conditions. RESULTS: While the mean or the SD of all the 56 points was not significantly different among the three conditions, APEN was highly and significantly lower for pemoline compared with fluoxetine and placebo. There was no significant correlation between average APEN and mean or SD (standard deviation). The one symptom that explained most of this difference among the groups after drug administration was the feeling of 'happiness'. CONCLUSIONS: This result indicates that there was a more consistent subjective sense of happiness during the 8-week period of pemoline administration compared with the other two drugs. Though this study was not designed to address the efficacy of mood stabilizing drugs, such daily subjective ratings may be useful in future studies that evaluate the stability of mood. APEN has been used in several different fields of research with small data sets and this study extends its possible use to evaluate changes in mood in certain populations such as patients with bipolar disorders. PMID- 12895206 TI - Quantitative assessment strategies and issues for mood and other psychiatric serial study data. PMID- 12895207 TI - Allelic association study between phospholipase A2 genes and bipolar affective disorder. AB - OBJECTIVES: In vivo studies demonstrating that lithium is a powerful phospholipase A2 (PLA2) inhibitor suggest that PLA2 activation, and subsequent cell signaling overactivation by increased fatty acid release may be the primary abnormality in bipolar affective disorder (BPAD), thus making PLA2 genes attractive candidates for the susceptibility to BPAD. The present study investigates polymorphisms in cytosolic phospholipase A2 (cPLA2), calcium independent phospholipase A2 (iPLA2), and secretory phospholipase (sPLA2) genes in a Brazilian sample. METHODS: A cross-sectional study was performed with 181 unrelated DSM-IIIR BPAD subjects and 312 controls. A polymerase chain reaction restriction fragment length polymorphism assay for BanI cPLA2 and AvrII iPLA2 polymorphisms was performed, and an ATT repeat in sPLA2 was assessed using a semiautomated genetic analyzer (ALFexpress). RESULTS: There was no significant difference observed in the allelic and genotypic distribution between the BPAD and control groups for cPLA2 (genotype: chi2 = 0.8, 2df, p = 0.6; allele chi2 = 0, 1df, p = 0.9), iPLA2 (genotype: chi2 = 1.7, 2df, p = 0.4; allele: chi2 = 0.3, 1df, p = 0.6), and sPLA2 (allele: chi2 = 3.6, 6df, p = 0.8). CONCLUSION: Our results failed to demonstrate that the studied PLA2 polymorphisms were associated with an increased risk for BPAD in our sample. PMID- 12895208 TI - Oral choline decreases brain purine levels in lithium-treated subjects with rapid cycling bipolar disorder: a double-blind trial using proton and lithium magnetic resonance spectroscopy. AB - OBJECTIVES: Oral choline administration has been reported to increase brain phosphatidylcholine levels. As phospholipid synthesis for maintaining membrane integrity in mammalian brain cells consumes approximately 10-15% of the total adenosine triphosphate (ATP) pool, an increased availability of brain choline may lead to an increase in ATP consumption. Given reports of genetic studies, which suggest mitochondrial dysfunction, and phosphorus (31P) magnetic resonance spectroscopy (MRS) studies, which report dysfunction in high-energy phosphate metabolism in patients with bipolar disorder, the current study is designed to evaluate the role of oral choline supplementation in modifying high-energy phosphate metabolism in subjects with bipolar disorder. METHODS: Eight lithium treated patients with DSM-IV bipolar disorder, rapid cycling type were randomly assigned to 50 mg/kg/day of choline bitartrate or placebo for 12 weeks. Brain purine, choline and lithium levels were assessed using 1H- and 7Li-MRS. Patients received four to six MRS scans, at baseline and weeks 2, 3, 5, 8, 10 and 12 of treatment (n = 40 scans). Patients were assessed using the Clinical Global Impression Scale (CGIS), the Young Mania Rating Scale (YRMS) and the Hamilton Depression Rating Scale (HDRS) at each MRS scan. RESULTS: There were no significant differences in change-from-baseline measures of CGIS, YMRS, and HDRS, brain choline/creatine ratios, and brain lithium levels over a 12-week assessment period between the choline and placebo groups or within each group. However, the choline treatment group showed a significant decrease in purine metabolite ratios from baseline (purine/n-acetyl aspartate: coef = -0.08, z = -2.17, df = 22, p = 0.030; purine/choline: coef = -0.12, z = -1.97, df = 22, p = 0.049) compared to the placebo group, controlling for brain lithium level changes. Brain lithium level change was not a significant predictor of purine ratios. CONCLUSIONS: The current study reports that oral choline supplementation resulted in a significant decrease in brain purine levels over a 12-week treatment period in lithium treated patients with DSM-IV bipolar disorder, rapid-cycling type, which may be related to the anti-manic effects of adjuvant choline. This result is consistent with mitochondrial dysfunction in bipolar disorder inadequately meeting the demand for increased ATP production as exogenous oral choline administration increases membrane phospholipid synthesis. PMID- 12895210 TI - Review article: the management of acute variceal bleeding. AB - Variceal haemorrhage is a common medical emergency with a high mortality (30 50%). Adequate resuscitation is vital, and once stabilised the patient should be moved to a high-dependency area. Antibiotics reduce mortality, and the vasoactive drug terlipressin should be administered if early endoscopy is unavailable. Early endoscopy is essential both to make the diagnosis and to allow therapeutic measures to be performed. The evidence suggests that variceal band ligation is the most effective therapy for oesophageal varices. If gastric varices are found at the index endoscopy the evidence at present is inadequate to be certain which is the best treatment, but both endoscopic therapy with cyanoacrylate or thrombin and transjugular intrahepatic portosystemic stent shunt (TIPSS) have been reported to be of benefit. When initial treatments fail, rescue therapy should be initiated. Most authorities agree that TIPSS is the rescue therapy of choice. Many questions remain concerning the treatment of acute variceal bleeding, particularly the ideal therapy for gastric varices and the role of combination vasoactive and endoscopic therapy. Randomised controlled trials are required to answer these important issues. PMID- 12895211 TI - Review article: mild to moderate Crohn's disease--defining the basis for a new treatment algorithm. AB - Previously, clinicians have had few choices in treating mild to moderate Crohn's disease. They currently treat these Crohn's disease patients with oral mesalamine and antibiotics. This treatment approach is based on the safety of these agents, and the perception that they are effective. This perception regarding efficacy may be influenced by publication bias. This review examines the efficacy and safety data of the conventional corticosteroids, mesalamine, sulfasalazine, budesonide and antibiotics for inducing the remission of mild to moderate Crohn's disease from randomized controlled trials, and proposes an evidence-based treatment approach. Sulfasalazine has demonstrated modest efficacy when Crohn's disease is confined to the colon. Mesalamine has no clear benefit over placebo in treating active Crohn's disease. Conventional corticosteroids effectively induce remission but are associated with unwanted adverse effects. Budesonide has similar efficacy to conventional steroids with far fewer adverse effects. Antibiotics have not consistently demonstrated efficacy. We propose a new evidence-based approach which suggests inducing remission of mild to moderate Crohn's disease with budesonide 9 mg/day for patients with ileal and/or right colonic involvement; sulfasalazine for those with disease limited to the colon; and conventional steroids for high disease activity, those who failed budesonide and those with left-sided disease who are allergic or intolerant to sulfasalazine. PMID- 12895212 TI - Meta-analysis: the relationship between Helicobacter pylori infection and gastro oesophageal reflux disease. AB - BACKGROUND: The relationship between Helicobacter pylori infection and its treatment and gastro-oesophageal reflux disease (GERD) is controversial. AIMS: To establish if H. pylori infection is associated with the presence of GERD and if anti-H. pylori treatment leads to de novo GERD or rebound/exacerbation of GERD. METHODS: A search of MEDLINE and EMBASE databases was made. Pooled odds ratios (OR) were calculated for de novo GERD and rebound/exacerbated GERD after anti-H. pylori therapy in case-control studies and in therapeutic trials. RESULTS: Fourteen case-control studies and 10 clinical trials were included. Among case control studies, pooled OR for the association between H. pylori negative status and GERD was 1.34 [95% confidence interval (CI) 1.15-1.55]. Among therapeutic trials, pooled OR for the association anti-H. pylori therapy - GERD was 2.54 (95% CI 1.92-3.37). The OR for de novo GERD was 3.25 (95% CI 2.09-5.33), and for rebound/exacerbated GERD was 2.39 (95% CI 1.75-3.34). Associations were higher among Asian studies than among North American and European studies. CONCLUSIONS: This meta-analysis shows significant association between absence of H. pylori infection and GERD symptoms, and a positive association between anti-H. pylori therapy and occurrence of both de novo and rebound/exacerbated GERD. The significance of these associations appears to have been inflated by the effect of single trials and by geographical variations. PMID- 12895213 TI - Contemporary understanding and management of reflux and constipation in the general population and pregnancy: a consensus meeting. AB - BACKGROUND: Gastro-oesophageal reflux disease (GERD) and constipation have a major impact on public health; however, the wide variety of treatment options presents difficulties for recommending therapy. Lack of definitive guidelines in pharmacy and general practice medicine further exacerbates the decision dilemma. AIMS: To address these issues, a panel of experts discussed the principles and practice of treating GERD and constipation in the general population and in pregnancy, with the aim of developing respective treatment guidelines. RESULTS: The panel recommended antacids 'on-demand' as the first-line over-the-counter treatment in reflux, and as rescue medication for immediate relief when reflux breaks through with proton pump inhibitors. Calcium/magnesium-based antacids were recommended as the treatment of choice for pregnant women because of their good safety profile. In constipation, current data do not distinguish a hierarchy between polyethylene glycol (PEG)-based laxatives and other first-line treatments, although limitations are associated with stimulant- and bulk-forming laxatives. Where data are available, PEG is superior to lactulose in terms of efficacy. In pregnancy, PEG-based laxatives meet the criteria for the ideal treatment. CONCLUSIONS: The experts developed algorithms that present healthcare professionals with clear treatment options and management strategies for GERD and constipation in pharmacy and general practice medicine. PMID- 12895214 TI - Long-term results of low-dose intravenous ciclosporin for acute severe ulcerative colitis. AB - BACKGROUND: Intravenous ciclosporin for acute, severe colitis is usually administered in a dose of 4 mg/kg/day, with concurrent intravenous steroids. This is associated with considerable morbidity. We have been using a low-dose regimen, most commonly without concurrent steroids, for seven years, and present the outcome. METHODS: Records of all patients admitted for severe ulcerative colitis, treated by one physician over seven years, were reviewed. RESULTS: Thirty-one patients received low-dose intravenous ciclosporin (2 mg/kg/day) for a median 8 days. Eleven early patients received concurrent intravenous corticosteroids. Three patients had hypertension requiring dose reduction, one elevated creatinine and one elevated liver enzymes (all transient), and four experienced infection (two arm cellulitis, one perianal abscess, one post-operative wound infection). Twenty-four patients (77%) avoided urgent colectomy, and were discharged on oral ciclosporin and azathioprine. After a median 18 months (range 3-77), 14 patients (45% of total) avoided colectomy, of whom eight had flares responding to medical therapy and two had persistent, mildly active disease. CONCLUSIONS: Low-dose intravenous ciclosporin (2 mg/kg/day), usually used as a monotherapy and followed by azathioprine, achieves similar long-term efficacy to higher dose ciclosporin combined with steroids in severe acute ulcerative colitis. Morbidity appears to be low. PMID- 12895215 TI - The mechanisms of prednisone inhibition of inflammation in Crohn's disease involve changes in intestinal permeability, mucosal TNFalpha production and nuclear factor kappa B expression. AB - BACKGROUND: The clinical course of Crohn's disease after the induction of remission with medical therapy is characterized by unpredictable relapse. AIM: To evaluate three surrogate markers, intestinal permeability, mucosal TNFalpha and nuclear factor (NF)-kappaB/IkappaBalpha expression, in order to determine the relationship of these parameters to clinical relapse. METHODS: Thirty patients with active Crohn's disease were treated with a 10 week course of prednisone using a tapering dosing regimen. Intestinal permeability (lactulose/mannitol [L/M ratio]) was determined at baseline and at the end of prednisone tapering. TNFalpha production and the levels of expression of NF-kappaB/IkappaBalpha were measured in colonic mucosal biopsies obtained after the induction of remission. RESULTS: Twenty-two patients (73%) achieved remission and 50% of patients experienced a clinical relapse during the ensuing 12 months. Treatment with prednisone resulted in a significant decrease in the L/M ratio. Of the patients that relapsed, 75% had a raised L/M ratio at the time of remission compared with 20% of patients with a normal L/M ratio (P < 0.008; hazard ratio = 6.094; CI 1.55, 17.43). Mucosal TNFalpha production was greater in relapsers compared with those who remained in remission. The levels of NF-kappaB in relapsers were significantly greater and levels of cytosolic IkappaBalpha were significantly lower compared with those measured in patients who remained in remission. CONCLUSIONS: These findings underscore the importance of incorporating biological parameters of inflammation in determining the clinical course of Crohn's disease. PMID- 12895216 TI - Coping strategies, illness perception, anxiety and depression of patients with idiopathic constipation: a population-based study. AB - BACKGROUND: Functional constipation has important psychological elements. AIM: To investigate the prevalence of functional constipation in an Asian population, and the interplay among functional constipation, anxiety/depression, perception and coping strategies. METHODS: An interview of 3282 patients was made by telephone survey. Constipation was diagnosed by Rome II criteria. Coping ability and anxiety/depression were assessed by validated questionnaires. RESULTS: Fourteen percent of the interviewees had constipation. Anxiety and depression scores were higher in constipated than in healthy subjects (P < 0.0001 and < 0.0001), and in female than male patients (P = 0.02 and < 0.0001). Patients who were aware of their symptoms perceived greater impact on their lives (P < 0.001). Frequent use of coping strategies associated with lower anxiety scores (P < 0.0001). Female were more frequently aware of the symptoms (P = 0.004), less frequently used coping strategies (P = 0.008). Regression analysis showed that female and high anxiety level were the independent factors for predicting the perception of constipation, whereas anxiety was the only independent factor for predicting the use of coping strategies. CONCLUSION: Constipation associated with anxiety and depression is prevalent in the general Asian population. Female sex and anxiety are important aetiological factors in constipation, affecting perception and the use of coping strategies. PMID- 12895217 TI - Pre-medication with pronase reduces artefacts during endoscopic ultrasonography. AB - BACKGROUND: Gastric mucus usually induces artefacts during endoscopic ultrasonography. AIM: To investigate the effectiveness of pre-medication with the proteolytic enzyme, pronase, before endoscopic ultrasonography. METHODS: Out patients scheduled for endoscopic ultrasonography were randomly assigned to oral pre-medication with the anti-foam agent, dimethylpolysiloxane, alone (treatment A; n = 29), with dimethylpolysiloxane plus sodium bicarbonate (treatment B; n = 29) or with dimethylpolysiloxane, sodium bicarbonate and pronase (treatment C; n = 29). All drinks were given about 10 min before the start of the procedure. After insertion of the endoscope, endoscopists recorded visibility scores before the procedure, imaging scores at endoscopic ultrasonography and the numbers of high-echo spots in the gastric cavity and on the gastric wall surface after the procedure. RESULTS: Pre-medication with pronase (treatment C) significantly reduced (both at P < 0.05) the visibility score (score 4, 46%) in comparison with that obtained for pre-medication without pronase (10% for both treatments A and B). Treatment with pronase significantly reduced (both at P < 0.05) the endoscopic ultrasonography score in the gastric cavity (score 4, 34%) in comparison with that found for treatments A (7%) and B (0%). It also significantly reduced (P < 0.05) the endoscopic ultrasonography score on the gastric wall surface (score 4, 14%) in comparison with that observed for treatment A (3%). The numbers of high-echo spots in the gastric cavity and on the gastric wall surface were significantly less (both at P < 0.001) for pre medication with pronase (treatment C) than for pre-medication with treatments A and B. There were no complications associated with the solutions. CONCLUSIONS: Pre-treatment with pronase reduced the artefacts during endoscopic ultrasonography. PMID- 12895218 TI - CCK-1 receptor blockade for treatment of biliary colic: a pilot study. AB - BACKGROUND: Loxiglumide is a potent and selective cholecystokinin-1 (CCK-1) receptor antagonist able to inhibit gall-bladder contraction. AIM: To assess the effect of CCK-1 receptor blockade on the pain of patients with biliary colic. PATIENTS AND METHODS: Fourteen patients with biliary colic but no suspicion for acute cholecystitis, were randomly and blindly assigned to loxiglumide (50 mg i.v.) or hyoscine-N-butyl bromide (20 mg i.v.) treatment. Pain intensity was monitored by a Visual Analogue Scale. Patients with less than 80% response at 30 min, were retreated with a second injection of the same compound. RESULTS: Reduction in pain score (mean +/- S.E.M.) was faster and significantly greater in patients treated with loxiglumide (n = 7) than in controls (n = 7): 88 +/- 7% vs. 47 +/- 12% after 20 min, P < 0.05; 92 +/- 6% vs. 49 +/- 13%, after 30 min, P < 0.05. Only one of seven patients treated with loxiglumide needed a second injection at 30 min (vs. six of seven controls, P < 0.05). No adverse effect was observed after either treatment. CONCLUSIONS: Loxiglumide is highly effective in obtaining pain relief in patients with biliary colic. The analgesic effect of CCK 1 receptor blockade is superior to that of a conventional anticholinergic treatment. PMID- 12895219 TI - Double-blinded placebo-controlled study of Phyllanthus urinaris for the treatment of chronic hepatitis B. AB - BACKGROUND: Previous studies suggested that Phyllanthus species have an anti viral effect on hepatitis B, but methodologies have been inadequate. AIMS: This study aimed to investigate the anti-viral effect of Phyllanthus urinaris. METHODS: Chronic hepatitis B patients with positive hepatitis B e-antigen (HBeAg), hepatitis B virus (HBV) DNA > 500 000 copies/mL and elevated alanine transaminase (ALT) were recruited. Patients were randomized into groups of 12 receiving P. urinaris 1, 2 and 3 g three times daily for 6 months or placebo (six cases). The primary endpoint was HBV DNA reduction, and secondary endpoints were HBeAg seroconversion and ALT normalization. RESULTS: On an intention-to-treat analysis there was no difference in log10[HBV DNA] reduction of the Phyllanthus 1 g (0.18 +/- 1.42), 2-g (0.33 +/- 1.08) and 3-g (0.85 +/- 1.30) groups vs. placebo (0.28 +/- 0.85) (P = 0.90, 0.92 and 0.38, respectively) at the end of treatment. The percentage of patients among the placebo, Phyllanthus 1-g, 2-g and 3-g groups undergoing HBeAg seroconversion (0%, 9.1%, 8.3% and 16.7%, respectively) and ALT normalization (0%, 0%, 8.3% and 33.3%) were not significantly different at the end of treatment. No delayed virological or biochemical response was documented at 24 weeks after the cessation of treatment. No serious adverse event was reported. CONCLUSION: P. urinaris treatment for 6 months has no demonstrable anti viral effect in chronic hepatitis B. PMID- 12895220 TI - Quadruple therapy containing amoxicillin and tetracycline is an effective regimen to rescue failed triple therapy by overcoming the antimicrobial resistance of Helicobacter pylori. AB - AIM: To identify optimal antibiotics for second-line quadruple therapy of Helicobacter pylori after failed 1-week triple therapy. METHODS: One hundred patients were enrolled in this study after the failure of 1-week triple therapy. They were randomized to receive 1-week quadruple therapy consisting of amoxicillin, omeprazole and bismuth salts, plus either metronidazole or tetracycline. Before quadruple therapy, the H. pylori culture of each patient was tested for metronidazole resistance or clarithromycin resistance by E-test. Six weeks later, an endoscopy or 13C-urea breath test was used to define the success of H. pylori eradication. RESULTS: The H. pylori eradication rates by intention to-treat and per protocol analysis were higher in the tetracycline group than in the metronidazole group (intention-to-treat: 78% vs. 58%, P < 0.05; per protocol: 89% vs. 67%, P < 0.05). In the metronidazole group, but not in the tetracycline group, the per protocol eradication rate of quadruple therapy was lower for the infected isolates with metronidazole resistance than for those without metronidazole resistance (77% vs. 33%, P < 0.05). CONCLUSION: Quadruple therapy, including tetracycline and amoxicillin, improves the H. pylori eradication rate after failed triple therapy. PMID- 12895222 TI - Evaluation of the clinical and allergen specific serum immunoglobulin E responses to oral challenge with cornstarch, corn, soy and a soy hydrolysate diet in dogs with spontaneous food allergy. AB - Fourteen dogs with known clinical hypersensitivity to soy and corn were maintained on a limited antigen duck and rice diet until cutaneous manifestations of pruritus were minimal (78 days). Sequential oral challenges with cornstarch, corn and soy were then performed. Subsequently, the dogs were fed a diet containing hydrolysed soy protein and cornstarch. Throughout the study period the dogs were examined for cutaneous manifestations of pruritus and, additionally, serum was collected for measurement of allergen-specific and total immunoglobulin (Ig)E concentrations. Intradermal testing with food antigens was performed prior to entry into the study and after 83 days. A statistically significant clinical improvement was measured between days 0 and 83. Significant pruritus was induced after oral challenge with cornstarch, corn and soy (P = 0.04, 0.002, 0.01, respectively) but not with the hydrolysed diet (P = 0.5). The positive predictive value of the skin test for soy and corn allergy was reduced after feeding a soy and corn free diet. Although increases in soy and corn-specific serum IgE concentrations were measured in individual dogs post challenge they were not statistically significant and could not be used to predict clinical hypersensitivity. PMID- 12895223 TI - Efficacy of milbemycin oxime in the treatment of canine generalized demodicosis: a retrospective study of 99 dogs (1995-2000). AB - Ninety-nine dogs diagnosed with generalized demodicosis were treated with milbemycin oxime. Cases diagnosed before two years of age were classified as juvenile-onset (53%) and those diagnosed after two years of age as adult-onset (47%). Dogs were considered cleared of mites when none could be demonstrated in scrapings and cured if no relapse was seen for 12 months. Eighty-five per cent (84/99) were cured with milbemycin oxime used for 1-6 months (mean 2.3 months) at a dosage of 0.5-1.6 mg kg-1 body weight (mean 0.75 mg kg-1). No significant difference in dosage or treatment time was seen between juvenile and adult cases. Chance of cure was significantly better in young animals (cured cases mean age 2.97 years) than in older animals (not cured cases mean age 8.02 years). Start of treatment early in the course of disease gave a significantly better chance of cure. Cases with severe pododemodicosis had less chance of a cure (9/11 not cured). PMID- 12895224 TI - Hereditary nasal parakeratosis in Labrador retrievers: 11 new cases and a retrospective study on the presence of accumulations of serum ('serum lakes') in the epidermis of parakeratotic dermatoses and inflamed nasal plana of dogs. AB - We report 11 new cases of hereditary nasal parakeratosis in Labrador retrievers. The disease was first observed when the dogs were 6 months to 2 years of age, and affected dogs of either sex and all coat colours. Hyperkeratosis and depigmentation were confined to the nasal planum, and affected dogs were otherwise healthy. The principal histological findings in biopsy specimens were marked diffuse parakeratotic hyperkeratosis, multiple intracorneal serum lakes and superficial interstitial-to-interface lymphoplasmacytic dermatitis. Topical applications of propylene glycol in water or white petrolatum were often effective for treatment of the dermatosis. However, continued applications were required to maintain a beneficial response. A retrospective histological study of parakeratotic inflammatory diseases of canine haired skin and inflammatory diseases of the canine nasal planum was performed. The degree of parakeratotic hyperkeratosis and the number and size of intracorneal serum lakes were evaluated. The degree of parakeratotic hyperkeratosis was greater in hereditary nasal parakeratosis specimens than that seen in discoid lupus erythematosus and Malassezia dermatitis. There were more serum lakes in hereditary nasal parakeratosis specimens than in specimens from dogs with discoid lupus erythematosus, Malassezia dermatitis, primary seborrheic dermatitis or zinc responsive dermatosis. Significant differences in sizes of serum lakes (if present) were not seen. PMID- 12895225 TI - Epidermal (infundibular) and dermoid cysts in the dorsal midline of a three-year old thoroughbred-cross gelding. AB - Epidermal (infundibular) and dermoid cysts are unusual in the horse in contrast with other species. The diagnosis and treatment of six lesions in the dorsal midline of a three-year-old Thoroughbred-cross gelding is described. The lesions were believed to be congenital and presented asymptomatically but required attention because five of them were in the saddle region, thus preventing ridden exercise. Under general anaesthesia, the cysts were excised and subsequently examined histologically. The horse recovered uneventfully. This report is novel in that such midline cysts have not previously been described outside Australia and North America. PMID- 12895227 TI - Prognostic significance of the infiltrative pattern invasion in endometrioid adenocarcinoma of the endometrium. AB - The prognostic significance of the invasive type of carcinoma cells in endometrial carcinoma is not defined. We evaluated the prognostic significance of the invasive type, as well as the immunostains of p53, c-erbB-2, Ki-67 antigen and MDM2 in endometrial endometrioid adenocarcinoma. This prospective analysis comprised 112 patients with endometrioid adenocarcinoma of the uterine corpus who had undergone surgery and were traced for more than 5 years after the operation. They were divided into recurrence (16 patients) and non-recurrence (96 patients) groups. The invasive type of carcinoma cells was divided into expansile, mixed (expansile and infiltrative) and infiltrative pattern. The difference in the invasive type (P < 0.001) and p53 expression (P = 0.004) between the recurrence and non-recurrence groups was significant in the univariate analysis. Moreover, the invasive type was significant in the multivariate analysis (P = 0.004). In contrast, the difference in MDM2 expression, c-erbB-2 expression and the Ki-67 labeling index in both groups was not significant in the univariate analysis. The infiltrative pattern of the invasive type (P < 0.001) and p53 expression (P = 0.043) were significantly related to a poor prognosis in the Kaplan-Meier method using the log-rank test. In conclusion, the current study indicated that the infiltrative pattern of the carcinoma cells is a predictor for poor prognosis in endometrioid adenocarcinoma in the uterine corpus. It was also indicated that p53 immunostains are useful as a predictor, but Ki-67 antigen, c-erbB-2 and MDM2 stains are not. PMID- 12895228 TI - Comedonecrosis is an unfavorable marker in node-negative invasive breast carcinoma. AB - Breast carcinoma is usually accompanied by an invasive component with an intraductal component, and each component shows different morphological features. We evaluated whether the presence or absence of comedonecrosis is correlated with prognosis and biological features in node-negative invasive breast carcinoma. Ninety-four node-negative breast carcinomas with an intraductal component were classified into two types: comedo type (n = 36) showing comedonecrosis partly or extensively in the intraductal component, and non-comedo type (n = 58) showing either an absence or small foci of necrosis. The Kaplan-Meier method was used to calculate disease-free survival. Immunohistochemical examination for p53 and HER 2 was conducted on the comedo (n = 35) and non-comedo (n = 47) type tumor specimens. Disease-free survival was significantly shorter in the comedo type than in the non-comedo type (P = 0.019). The expression of p53 was observed in 16 (45.7%) of the 35 comedo type cases, but only in two (4.3%) of the 47 non-comedo type cases (P < 0.0001). HER-2 overexpression was observed in seven (20.0%) of the 35 comedo type cases, while none of the 47 non-comedo type cases overexpressed HER-2 (P < 0.0001). These results suggest that the presence of comedonecrosis may be predictive of an unfavorable prognosis with aggressive biological behavior in node-negative invasive breast carcinoma. PMID- 12895229 TI - Glomerulosclerosis develops in Thy-1 nephritis under persistent accumulation of macrophages. AB - To clarify the relationship between macrophages and development of glomerulosclerosis, the authors developed a new experimental nephritis model with macrophages persisting in Thy-1 nephritis. Methyl-cellulose was administered intraperitoneally in addition to the intravenous injection of the anti-Thy-1 antibody to Wistar rats. Foamy macrophages influxed into the lytic mesangium and stayed to form nodular aggregates. Mesangial cells proliferated with the formation of extracellular matrices around these nodular aggregates of macrophages. Immunohistochemical analyses revealed that alpha-smooth muscle actin (alpha-SMA) was expressed in the proliferative area around these nodules of foamy macrophages from day 7. Type I collagen and type IV collagen were also expressed around the foamy macrophages in correspondence with alpha-SMA expression from day 7. The electron microscopic study revealed that collagen fibrils were formed around the transformed mesangial cells. The expression of platelet endothelial cell adhesion molecule-1 (PECAM-1, CD31), a marker of glomerular vasculature endothelial cells, was not found in the area occupied by the foamy macrophages, suggesting the impairment of glomerular reconstruction. Macrophages may participate in the progression of glomerulosclerosis in Thy-1 nephritis by enhancing the production of the extracellular matrix through transformed mesangial cells and preventing reconstruction of the capillary network. PMID- 12895230 TI - Bone marrow cell transfer of autoimmune diseases in a MRL strain of mice with a deficit in functional Fas ligand: dissociation of arteritis from glomerulonephritis. AB - MRL/MpTn-gld/gld (MRL/gld) mice, which are deficient in a functional Fas ligand (FasL), spontaneously develop autoimmune diseases involving both lethal glomerulonephritis and systemic arteritis, while MRL/Mp-+/+ (MRL/+) and C3H/HeJ gld/gld (C3H/gld) do not. To determine the cells responsible for the development of glomerulonephritis and arteritis, we transferred bone marrow cells from MRL/gld mice to undiseased MHC-compatible gld/gld or +/+ mice. In bone marrow irradiation chimeras, MRL/gld bone marrow cells were transferred to lethally irradiated MRL/+ or C3H/HeJ-+/+ (C3H/+) mice, and both recipients developed glomerulonephritis associated with hypergammaglobulinemia without causing graft versus-host (GVH)-like diseases. However, a striking difference between them was that MRL/+ recipients developed arteritis, but C3H/+ recipients did not. In bone marrow mixed chimeras formed by transferring MRL/gld bone marrow cells to unirradiated mice, the MRL/gld bone marrow cells induced glomerulonephritis in C3H/gld mice, but not in C3H/+ and MRL/+ mice. These results indicate that bone marrow cells from MRL/gld mice can cause glomerulonephritis in mice, even in those with a C3H background, possibly if they survive longer by escaping from Fas mediated apoptosis, while the development of arteritis requires the MRL genetic background in the recipients. This is the first report of the transfer of arteritis in lupus mice to undiseased recipients. PMID- 12895231 TI - Cross-reactive antigenicity of nucleoproteins of lyssaviruses recognized by a monospecific antirabies virus nucleoprotein antiserum on paraffin sections of formalin-fixed tissues. AB - Diagnosis of rabies is routinely confirmed by detection of rabies virus antigens in acetone-fixed frozen brain tissues or imprint smears using an immunofluorescence method with commercial antirabies virus antibodies. Since recent molecular analyses disclosed wide heterogeneity in the genome sequences of rabies virus strains and related lyssaviruses, it is necessary to confirm the presence of common epitopes in these lyssaviruses. In this study we confirmed the presence of cross-reactive antigens of various lyssaviruses in paraffin sections of formalin-fixed tissue using a monospecific rabbit antiserum prepared by immunization with a recombinant nucleoprotein of rabies virus. By immunohistochemical application, the antigen was detected predominantly in the cytoplasm of neurons in the brains of mice infected with rabies virus, Duvenhage virus, Mokola virus and European bat lyssavirus-1, while no cross-reaction was observed in uninfected humans and animals including dogs, bats, and raccoons. In addition, we examined one autopsy case that was infected in a rabies-endemic nation and developed the clinical manifestation of rabies after returning to Japan in 1970, and found that the antigen was well preserved in paraffin sections of formalin-fixed tissues. Thus, this suggests that the lyssavirus-specific antigen is recognized by the monospecific antibody against rabies virus nucleoprotein, and that this cross-reactive antigen is detectable on formalin fixed paraffin-embedded tissues by immunohistochemical analysis. PMID- 12895232 TI - Diagnosis of renal amyloidosis using Congo red fluorescence. AB - Early diagnosis and classification of amyloid deposition and differentiation from other glomerular fibrillar deposits relies on routine Congo red (CR) histochemistry. Congo red fluorescence (CRF) is an alternative method based on examination of the CR-stained section by ultraviolet (UV) light. The aim of this study is to investigate the usefulness of CRF, especially when applied to frozen kidney sections. Congo red fluorescence was applied to sections of frozen kidney biopsies prospectively and to paraffin sections retrospectively. The findings of CRF were compared to CR staining in bright light. Prospectively, 15 cases of amyloidosis were diagnosed on frozen sections and identical CR staining was found in all of the paraffin-stained sections. There were no false positives or negatives. Retrospectively, 146 renal biopsies previously stained with CR were re evaluated with CRF. Eighty-seven CR positive cases were confirmed by CRF, and one new case was identified. Congo red fluorescence is simple to perform and more pronounced, therefore easier to evaluate than CR in bright light. Congo red, when combined with immunohistochemistry, is still visible under UV whereas CR is masked in bright light. Although not widely used, the CRF method for detecting amyloid is simple to use with a high specificity and sensitivity, and may be applied successfully to frozen sections. PMID- 12895233 TI - Highly anaplastic extraventricular ependymoma arising in an adult, mimicking metastatic adenocarcinoma with heavy stromal inflammation and emperiporesis. AB - We report a case of extraventricular ependymoma arising in a 50-year-old woman that took an aggressive clinical course with recurrence three times. The initial tumor was a well-circumscribed nodule in the right temporal white matter measuring 2 cm in diameter. It showed variegated histological findings mimicking metastatic adenocarcinoma: an epithelioid arrangement of highly pleomorphic cells with pseudopapillary structures and perivascular pseudorosettes, and bizarre multinucleated giant cells with occasional emperiporesis surrounded by abundant mononuclear inflammatory cells, as well as a focal small area of conventional ependymoma. Emperiporesis and abundant mononuclear cell infiltration were not previously described in an ependymoma. The recurrent tumors predominantly showed an epithelioid pattern with frequent formation of astroblastoma-like pseudopapillary structures. Neoplastic cells were markedly atypical and had characteristic intracytoplasmic eosinophilic inclusion bodies. Much of the cells in both the initial and recurrent tumors showed a positive immunostaining for glial fibrillary acidic protein (GFAP) with accentuation to the cytoplasmic processes of the pseudorosettes and pseudopapillary structures. Epithelial membrane antigen (EMA) highlighted the epithelial differentiation of the tumor cells, while cytokeratin was completely negative. Although this tumor might be classified to at least WHO grade III from the histology and aggressive behavior, the exact grading is still controversial because of the rarity of such cases. PMID- 12895234 TI - Pyothorax-associated angiosarcoma of the pleura with metastasis to the brain. AB - Pleural angiosarcoma is an extremely rare, highly malignant neoplasm. Chronic tuberculous pyothorax is one of the etiological factors associated with the development of pleural angiosarcoma. This report details a case of pleural angiosarcoma in a 70-year-old woman with a history of tuberculous pyothorax. Coagulated blood surrounded by thickened pleura in the right thorax and hematoma like multiple metastases in the brain were noted on autopsy. The pleural lesion was presumed to be the primary site. Microscopic examination revealed rudimentary channels lined by plump neoplastic cells in the coagulated blood of the pleura and the brain. These neoplastic cells stained positive for endothelial markers. A literature review of English language journals revealed this to be the first patient described in detail who developed cerebral metastasis secondary to pleural angiosarcoma. PMID- 12895235 TI - Carcinosarcoma (pure endocrine cell carcinoma with sarcoma components) of the stomach. AB - A case of gastric carcinosarcoma with a rare histology is reported. Grossly, a huge ulcerative tumor (Borrmann type 3 tumor, measuring 10 x 6 cm) was observed on the lesser curvature side of the stomach. Microscopically, the tumor consisted of carcinoma and sarcoma components. The main tumor component consisted of an endocrine cell carcinoma exhibiting positive reactions for cytokeratins, chromogranin A, synaptophysin, and CD56. The sarcoma component occupied less than 10% of the entire tumor area and consisted of spindle cells, some of which showed a rhabdomyosarcoma differentiation. Some of the spindle cells also showed an atypical cartilage or osseous differentiation. This sarcoma component exhibited positive staining not only for vimentin and desmin, but also for cytokeratins. Transitions between these two components were occasionally observed. These findings suggest that the sarcoma component in our case most likely originates from the endocrine cell carcinoma component. PMID- 12895236 TI - Discordant lymphoma: MALT lymphoma of the stomach and follicular lymphoma of the parotid gland. AB - More than one histological type of malignant lymphoma can occur simultaneously in an individual. The entity is classified as either composite or discordant lymphoma. Both types of lymphoma, particularly discordant lymphoma comprised of extranodal marginal zone B-cell lymphoma of mucosa-associated lymphoid tissue lymphoma (MALT-L) and follicular lymphoma (FL), are rare. We report a case of discordant lymphoma comprising MALT-L in the stomach and FL in the parotid gland. The patient was a 50-year-old Japanese woman who visited the University Hospital of Showa (Tokyo, Japan) because a barium study showed erosive gastric lesions. A gastro-intestinal endoscopy was performed 2 months after the barium study, which showed irregular erosions throughout the stomach body. A gastric biopsy showed MALT-L, and Helicobacter pylori (H. pylori) infection was confirmed. The patient had noticed a painless and elastic hard tumor mass of about 2 cm in diameter in the area of the left parotid gland 6 months before the barium study. We removed the parotid gland tumor and diagnosed it as FL 6 months after the barium study. We were able to diagnose the MALT-L and FL by morphological, immunohistochemical and molecular analyses of paraffin-embedded sections. This appears to be the first reported case of MALT-L and FL occurring together as a discordant lymphoma. PMID- 12895237 TI - Marked diffuse dilations of the biliary tree associated with intrahepatic calculi, biliary sludges and a mucinous cyst of the pancreatic head in a 99-year old woman. AB - A 99-year-old woman was admitted to Shizuoka Shimizu Municipal Hospital because of fever and anasarca. Imaging and laboratory tests showed pneumonia, urinary tract infection, and cardiac failure. The patient died 20 days after admission. An autopsy revealed marked diffuse dilations of the biliary tree ranging from the lower common bile duct to intrahepatic bile ducts. Intrahepatic calcium bilirubinate stones and biliary sludges were recognized within the dilated bile ducts. A unilocular cyst (2 cm in diameter) was present in the pancreatic head adjacent to the lower common bile duct, and it appeared to compress the common bile duct. Histologically, the walls of the dilated biliary tree showed proliferation of peribiliary glands, fibrosis, and infiltration of lymphocytes and neutrophils (cholangitis). The lumens of the dilated biliary ducts contained neutral and acidic mucins, fibrinous materials, bacteria, neutrophils, and Aspergillus fungi, in addition to the calculi and sludges. The background liver showed atrophy (400 g). The pancreatic unilocular cyst was composed of mucous columnar cells with a few infoldings, and the pancreas also showed foci of mucinous duct hyperplasia and ectasia; the pathological diagnosis of the cyst was cystic dilations of a pancreatic duct branch (mucinous ductal ectasia or mucinous cyst). Other lesions included aspiration pneumonia, emaciation, atrophy of systemic organs, gastric leiomyoma, serous cystadenoma of the right ovary, and arteriosclerotic nephrosclerosis. The present case suggests that a mucinous cyst of the pancreas may compress the biliary tree and lead to marked diffuse dilations of the biliary tree. Alternatively, the dilations of the bile ducts may be associated with aging or may be of congenital origin. The dilated bile ducts may, in turn, give rise to bacterial and fungal cholangitis and formation of biliary sludges and intrahepatic calcium bilirubinate stones. PMID- 12895238 TI - Polyangitis overlap syndrome: a fatal case combined with adult Henoch-Schonlein purpura and polyarteritis nodosa. AB - Henoch-Schonlein purpura (HSP) is a rather common disease characterized by systemic hypersensitivity vasculitis in the skin and other visceral organs. It has a favorable prognosis unless it is complicated by severe glomerular disease. We report a distinctive fatal case of systemic vasculitis combined with HSP and polyarteritis nodosa (PN) in a 56-year-old man who died of progressive renal failure one month after the onset of the disease. He complained of arthralgia, purpura of both lower extremities, nasal bleeding and tarry stool, and acute renal failure was noted at the time of admission to hospital. A skin biopsy from the purpura lesion exhibited leucocytoclastic vasculitis with IgA deposition, and HSP was considered. However, renal failure progressed rapidly and subsequently was complicated by acute myocardial infarction. Postmortem examination revealed PN type necrotizing vasculitis in the kidneys, heart and mesentery resulting in acute multiple infarctions of these organs. We think the current case was a polyangitis overlap syndrome. It is important to suspect the polyangitis overlap syndrome positively when progressive acute renal failure is seen in a patient with HSP and to begin appropriate therapy immediately. PMID- 12895239 TI - Intrarenal epidermal cyst. AB - An epidermal cyst is a very unusual cause of a renal mass, with only three cases reported in English literature. The authors report a case of this in a 51-year old man with left flank pain and hematuria. A 5 x 4 cm-sized cystic mass, having an irregular margin with stippled calcification, was identified in the lower pole of the left kidney. A retroperitoneal laparoscopic simple nephrectomy was performed under the assumption that it was a renal tumor. Histologically, the tumor was identical in appearance to an epidermal cyst within the skin. Aberrant ectodermal implantation during embryogenesis has been postulated as a histogenesis of intrarenal epidermal cyst. However, the present case had a history of renal stone, which was treated with extracorporeal shock wave lithotripsy. Therefore, the authors suspected the pathogenetic mechanism of this lesion was a metaplasia of traumatic origin. The diagnosis of an epidermal cyst was not made preoperatively in the present case, or in the cases described in the literature. This diagnosis should be included in the differentiation of calcified intrarenal masses. PMID- 12895241 TI - Lessons to be learned from questioning the "big assumptions". PMID- 12895242 TI - Guessing on selected-response examinations. PMID- 12895243 TI - Training junior doctors in complex skills. PMID- 12895244 TI - Hormone surge. PMID- 12895245 TI - Doggie diagnosis, diagnostic success and diagnostic reasoning strategies: an alternative view. PMID- 12895246 TI - Instituting a medical education revolution. PMID- 12895247 TI - Making the most of learning in the operating theatre: student strategies and curricular initiatives. AB - BACKGROUND: Students in medical programmes around the world are required to attend with their patient in the operating theatre, yet little is known about this area of medical education. Most of the published literature on the operating theatre is normative in character, written from the perspective of the surgeon as teacher. This paper, in contrast, reports the student's perspective as learner. It is an interpretive case study conducted in the department of surgery at an Australian university. METHODS: Research approaches were combined in a multi method strategy which included in-depth interviews with students and surgeons, observations in operating theatres, group interviews and a student survey. The aim was to understand the student's experience of the theatre as a learning environment, with a view to improving teaching and learning in this setting. FINDINGS: The operating theatre is a challenging place in which to learn. The challenges are conceptualized in this paper around three key domains: the challenge posed by the physical environment and the emotional impact of surgery as work; the challenge of the educational task, and the challenge of managing the social relations of work in the operating theatre. CONCLUSION: Students who report finding the operating theatre a useful experience are those who adopt active strategies to successfully manage their learning across the three domains. The paper describes curricular initiatives that have been introduced in a department of surgery to help more students make the most of their learning in theatres, and goes on to discuss the wider implications of the findings for surgical education beyond this case study. PMID- 12895248 TI - The transition from knowing to doing: teaching junior doctors how to use insulin in the management of diabetes mellitus. AB - OBJECTIVE: To develop and evaluate a short education programme to improve the skills and confidence of junior doctors in managing the glycaemic control of inpatients with diabetes mellitus. METHODS: A total of 15 junior doctors completed two 1-hour workshops on the practical skills required to manage the glycaemic control of insulin-treated patients. The workshops were based on simulated case scenarios presented in a workbook format. Pre-workshop performance and levels of confidence were tested, using a set of tasks matched to the learning objectives. Participants were re-tested immediately after the second workshop and again after 3 months. RESULTS: There was a significant overall effect for time of testing for performance and confidence considered together, F(4,11) = 12.67, P = 0.000, power = 1.00. The mean score for performance for the intermediate and 3-month post-tests combined was significantly higher than the mean performance score for the pre-test (11.00 < [17.53 + 15.80]), t(56) = -6.50, P = 0.000 (95% CI -6.15, -3.10). The mean score for confidence for the intermediate and 3 month post-tests combined was higher than the mean for the pre test (13.20 < [15.33 + 15.20]), t(56) = 2.95, P = 0.011 (95% CI 2.19, 0.46), although this result must be treated with caution. CONCLUSIONS: A brief educational intervention can improve and maintain the performance and confidence of junior doctors in managing patients with insulin-treated diabetes in a simulated environment. PMID- 12895249 TI - Diagnostic reasoning strategies and diagnostic success. AB - PURPOSE: Cognitive psychology research supports the notion that experts use mental frameworks or "schemes", both to organize knowledge in memory and to solve clinical problems. The central purpose of this study was to determine the relationship between problem-solving strategies and the likelihood of diagnostic success. METHODS: Think-aloud protocols were collected to determine the diagnostic reasoning used by experts and non-experts when attempting to diagnose clinical presentations in gastroenterology. RESULTS: Using logistic regression analysis, the study found that there is a relationship between diagnostic reasoning strategy and the likelihood of diagnostic success. Compared to hypothetico-deductive reasoning, the odds of diagnostic success were significantly greater when subjects used the diagnostic strategies of pattern recognition and scheme-inductive reasoning. Two other factors emerged as independent determinants of diagnostic success: expertise and clinical presentation. Not surprisingly, experts outperformed novices, while the content area of the clinical cases in each of the four clinical presentations demonstrated varying degrees of difficulty and thus diagnostic success. CONCLUSIONS: These findings have significant implications for medical educators. It supports the introduction of "schemes" as a means of enhancing memory organization and improving diagnostic success. PMID- 12895250 TI - Improving medical student performance in adolescent anti-smoking health promotion. AB - BACKGROUND: We developed a 1-hour teaching seminar for medical students on anti smoking health promotion for adolescents. We report the development of the programme and a performance-based evaluation. AIM: To develop a seminar for medical students with the objective of improving medical student inquiry into smoking and the delivery of advice accordingly for adolescent patients in routine consultations. METHOD: The seminar was developed using principles of adult learning and delivered to a cohort of medical students (intervention group). Participants were Year 5 medical students in their paediatrics term. A comparison group of 58 medical students who did not receive the seminar was studied in the semester prior to the intervention. The primary outcome measure was a change in anti-smoking health promotion practice within an objective structured clinical examination (OSCE), where taking a smoking history and making links between smoking status and health were indicated in a 15-year-old simulated patient (SP) with poorly controlled asthma. This was part of the summative assessment in the paediatrics term. RESULTS: In the OSCE, 21 of 58 (36%) students in the comparison group asked the asthmatic SP about smoking, compared with 41 of 56 (73%) students in the intervention group (P = 0.0001). In all, 15 (26%) of the comparison group students addressed the patient's smoking, compared with 33 of the 56 (59%) intervention group students (P = 0.0004). CONCLUSION: A carefully designed seminar addressing adolescent smoking can increase the frequency of smoking inquiry and advice by medical students. PMID- 12895251 TI - Training needs in adolescent medicine of practising doctors: a Swiss national survey of six disciplines. AB - OBJECTIVE: To assess and compare the training needs in adolescent medicine of doctors within 6 specialties as a basis for the development of pre/postgraduate and continuing medical education (CME) training curricula. DESIGN: Cross sectional postal survey. SETTING: Switzerland. PARTICIPANTS: National, representative, random sample of 1857 practising doctors in 6 disciplines (general practitioners, paediatricians, gynaecologists, internists, psychiatrists, child psychiatrists) registered with the Swiss Medical Association. MAIN OUTCOME MEASURES: Perceived importance of and training interest in 35 topics related to adolescent medicine listed in a self-administered, anonymous questionnaire. RESULTS: A total of 1367 questionnaires were returned, representing a response rate of 73.9%. Clear interest in adolescent medicine was reported by 62.1% of respondents. Topics perceived to be the most important in everyday practice were functional symptoms (71.4%), acne (67.1%), obesity (64.6%), depression-anxiety (68.1%) and communication with adolescents (61.7%). Differences between disciplines were especially marked for gynaecologists, who expressed interest almost exclusively in medical topics specific to their field. In contrast, other disciplines commonly reported a keen interest in psychosocial problems. Accordingly, interest in further training was expressed mostly for functional symptoms (62.4%), eating disorders (56.3%), depression-anxiety (53.7%) and obesity (52.6%). Issues related to injury prevention, chronic disease and confidentiality were rated as low priorities. CONCLUSIONS: Regardless of discipline, Swiss primary care doctors expressed a strong interest in adolescent medicine. Continuing medical education courses should include both interdisciplinary courses and discipline-specific sessions. Further training should address epidemiological and legal/ethical issues (e.g. injury prevention, confidentiality, impact of chronic conditions). PMID- 12895252 TI - Questioning the "big assumptions". Part I: addressing personal contradictions that impede professional development. AB - BACKGROUND: The ultimate success of recent medical curriculum reforms is, in large part, dependent upon the faculty's ability to adopt and sustain new attitudes and behaviors. However, like many New Year's resolutions, sincere intent to change may be short lived and followed by a discouraging return to old behaviors. Failure to sustain the initial resolve to change can be misinterpreted as a lack of commitment to one's original goals and eventually lead to greater effort expended in rationalizing the status quo rather than changing it. OBJECTIVE: The present article outlines how a transformative process that has proven to be effective in managing personal change, Questioning the Big Assumptions, was successfully used in an international faculty development program for medical educators to enhance individual personal satisfaction and professional effectiveness. This process systematically encouraged participants to explore and proactively address currently operative mechanisms that could stall their attempts to change at the professional level. CONCLUSIONS: The applications of the Big Assumptions process in faculty development helped individuals to recognize and subsequently utilize unchallenged and deep rooted personal beliefs to overcome unconscious resistance to change. This approach systematically led participants away from circular griping about what was not right in their current situation to identifying the actions that they needed to take to realize their individual goals. By thoughtful testing of personal Big Assumptions, participants designed behavioral changes that could be broadly supported and, most importantly, sustained. PMID- 12895253 TI - Questioning the "big assumptions". Part II: recognizing organizational contradictions that impede institutional change. AB - BACKGROUND: Well-designed medical curriculum reforms can fall short of their primary objectives during implementation when unanticipated or unaddressed organizational resistance surfaces. This typically occurs if the agents for change ignore faculty concerns during the planning stage or when the provision of essential institutional safeguards to support new behaviors are neglected. Disappointing outcomes in curriculum reforms then result in the perpetuation of or reversion to the status quo despite the loftiest of goals. Institutional resistance to change, much like that observed during personal development, does not necessarily indicate a communal lack of commitment to the organization's newly stated goals. It may reflect the existence of competing organizational objectives that must be addressed before substantive advances in a new direction can be accomplished. OBJECTIVE: The authors describe how the Big Assumptions process (see previous article) was adapted and applied at the institutional level during a school of medicine's curriculum reform. Reform leaders encouraged faculty participants to articulate their reservations about considered changes to provided insights into the organization's competing commitments. The line of discussion provided an opportunity for faculty to appreciate the gridlock that existed until appropriate test of the school's long held Big Assumptions could be conducted. CONCLUSIONS: The Big Assumptions process proved useful in moving faculty groups to recognize and questions the validity of unchallenged institutional beliefs that were likely to undermine efforts toward change. The process also allowed the organization to put essential institutional safeguards in place that ultimately insured that substantive reforms could be sustained. PMID- 12895254 TI - Medical students proposing questions for their own written final examination: evaluation of an educational project. AB - BACKGROUND: In order to emphasise learning more than control, from autumn 2000 we have invited medical students to propose questions for their own written examination in family medicine. One out of three student's proposals was guaranteed to be a part of their coming written examination, possibly somewhat modified. AIM: To evaluate how sixth year medical students experienced the project, and to what extent their performance in the examination was influenced. PARTICIPANTS: Sixth year medical students. MAIN OUTCOME MEASURES: The project was evaluated using (i) marks in examination; (ii) scores on self-administered questionnaires; and (iii) students' free text evaluation. RESULTS: Fifty-seven of 64 (89%) students taking their examination in autumn 2000, and 56 of 59 (95%) students taking the exam in spring 2001, responded. In autumn 2000, 34 (60%) students reported that the project had changed their learning strategies. During spring 2001, 46 of 56 students participated in producing questions, using a mean of 2.6 hours on the work. Students got 5-7% higher marks on their own questions on a scale ranging from 1 to 12. The students' free text evaluation showed that they had prepared especially thoroughly for the topics proposed by the students. They found it comforting to know at least one of the questions in the examination, and the students' questions were found relevant for general practice. CONCLUSION: Encouraging students to write questions for their own examination makes them feel more confident during the examination period, and may increase their reflective learning, without seriously limiting topics studied or violating the control function of the examination. PMID- 12895255 TI - Equity in medical school admissions. PMID- 12895256 TI - Stewart Mennin. PMID- 12895259 TI - Alternative medicines and the liver. AB - Complementary and Alternative Medicine (CAM) is a heterogeneous group of theories and practices that are becoming increasingly popular in the West - between 20% and 65% of patients use CAM on a regular basis. In the UK, CAM is provided by over a third of general practitioners as well as by hospitals. The subject of much debate in both the lay and medical press, CAM is subject to increasing scrutiny from clinical research. In this review, we discuss the available evidence for herbal medicines, including that for silymarin, glycyrrhizin, Chinese herbal medicines and other herbal mixtures. We also review evidence regarding the safety of herbal medicines, both in terms of hepatotoxicity and drug interactions. We conclude that although CAM may be of benefit in the treatment of liver disease, the available evidence is insufficient to recommend any of the available therapies. CAM has not yet been well studied in liver disease and rigorous evaluation with well-designed double-blind randomised controlled trails is required. Doctors need to be aware of the widespread use of CAM, ask their patients specifically regarding their use of CAM and be aware of the potential for hepatotoxicity and interactions. PMID- 12895260 TI - Beta-glucuronidase inhibitor tectorigenin isolated from the flower of Pueraria thunbergiana protects carbon tetrachloride-induced liver injury. AB - BACKGROUND/AIM: To understand the relationship between the fluctuation in serum beta-glucuronidase and hepatotoxicity, an inhibitor of beta-glucuronidase was isolated from the flowers of Pueraria thunbergiana and its hepatoprotective activity was measured. METHOD: Tectorigenin was isolated from the flowers of pueria thunbergiana as an inhibitor of beta-glucuronidase, and serum ALT, AST and biological parameters as markers for its hepatoprotective activity were measured on CCl4-induced liver injury in mice. The relationship between serum beta glucuronidase and hepatoprotective activities in mice was measured. RESULTS: : When tectorigenin at a dose of 100 mg/kg was intraperitoneally administered on CCl4-induced liver injury in mice, it significantly inhibited the increase of plasma ALT, AST and LDH activities. The inhibitory effect of tectorigenin is much more potent than that of dimethyl diphenyl bicarboxylate (DDB), which has been used as a commercial hepatoprotective agent. When tectoridin transformed to tectorigenin by intestinal bacteria was orally administered to mice, it showed hepatoprotective activity. However, when tectoridin was intraperitoneally administrated to mice, it did not exhibit hepatoprotective activity. Moreover, orally administered tectoridin not only inhibited beta-glucuronidase but also increased GSH content and GST activity on CCl4-induced hepatotoxicity of mice. CONCLUSION: We insist that an inhibitor of beta-glucuronidase tectorigenin may be hepatoprotective and tectoridin should be a prodrug transformed to tectorigenin. PMID- 12895261 TI - Clinical significance of antibody against hepatitis B virus core antigen in patients with hepatitis C virus-related hepatocellular carcinoma. AB - PURPOSE: We investigated the unsettled issue of whether seropositivity for antibody to hepatitis B core antigen (anti-HBc) affects characteristics of hepatitis C virus (HCV)-related hepatocellular carcinoma (HCC). METHODS: Antibody status was determined by enzyme immunoassay in 243 patients with this cancer, and associations with clinicopathologic characteristics and outcome were analysed. Serum hepatitis B virus (HBV) DNA was determined by real-time polymerase chain reaction. RESULTS: Of 235 patients with unequivocal serologic status, 142 were seropositive and 93 were seronegative. Clinicopathologic characteristics and overall cumulative survival rates were comparable between the two groups. However, seropositivity tended to predict poor outcome for patients in Child class B or C (P=0.068), those in tumour-nodes-metastasis-based stage 3 or 4 (P=0.081), those with tumours exceeding 25 mm (P=0.068), and those with a past history of clinical liver disease (P=0.088). Multivariate analysis identified serum albumin, portal vein tumour thrombosis, and tumour size as independent determinants of survival. Serum HBV DNA was below 1.7 log copies/ml in all 40 patients tested. CONCLUSIONS: Overall, the clinical features of HCV-HCC were unaffected by seropositivity for anti-HBc. Seropositivity tended to worsen prognosis for subgroup with poor hepatic reserve or advanced tumours. PMID- 12895262 TI - Effect of iron overload and dietary fat on indices of oxidative stress and hepatic fibrogenesis in rats. AB - BACKGROUND/AIMS: Oxidative stress is presumed to play an important role in hepatic fibrogenesis. Diets high in polyunsaturated fatty acids (PUFA) enhance fibrosis and have been associated with increased oxidative damage in some models of liver injury. The aim of this study was to determine the effects of dietary fat of varying PUFA content on iron-induced oxidative stress and fibrosis. METHODS: Rats were given parenteral iron and diets supplemented with coconut oil, safflower oil or menhaden oil. RESULTS: Hepatic iron overload was associated with induction of heme oxygenase-1, a sensitive indicator of oxidative stress, and with modest increases in hydroxyproline and procollagen I mRNA levels without histologically evident fibrosis, all of which were unaffected by dietary fat. In addition, iron loading was associated with increases in cysteine, gamma glutamylcysteine and glutathione. Dietary fat brought about the expected alterations in peroxidizability, but did not alter indices of oxidative damage. CONCLUSION: These data highlight the distinction between oxidative stress and oxidative damage and suggest that the former is not sufficient to elicit overt fibrosis. Furthermore, while hepatic iron overload leads to oxidative stress, there is an associated upregulation of antioxidant defenses involving thiol metabolism that may be a critical factor limiting the accumulation of oxidative damage. PMID- 12895263 TI - Prolongation of QTc interval: relationship with etiology and severity of liver disease, mortality and liver transplantation. AB - BACKGROUND: A prolonged QTc interval has been reported in patients with liver disease. The objectives of our study were to determine whether a prolonged QTc interval was an independent predictor of mortality in patients with cirrhosis and to examine the effect of liver transplantation (LT) on QTc interval. PATIENTS AND METHODS: We retrospectively studied two cohorts of patients - QTc interval was measured in 409 patients (pre-transplant group), and in 162 patients (transplant group) before and 6 months after LT. QT interval (mean) corrected (QTc) for ventricular rate was read from a 12-lead EKG. Patients with known cardiovascular disease or other risk factors that are known to cause a prolonged QTc interval were excluded. RESULTS: Pre-transplant group. One hundred and sixty-two patients (40%) had a prolonged QTc interval (>440 ms). By binary logistic regression, age (P=0.005), alcoholic cirrhosis (P=0.007) and Child-Pugh scores (P=0.007) were independent predictors of prolonged QTc interval. Sixty-six patients died during a mean follow-up of 8.9 years. Although the Kaplan-Meier survival curve showed a lower survival in patients with a prolonged QTc interval (P=0.03 by log rank test), when survival was adjusted for the Child-Pugh score by Cox regression survival analysis, there were no survival differences in patients with and without prolonged QTc interval. Cox regression analysis showed that the Child Pugh score (hazard ratio 1.5, CI 1.3-1.6, s<0.001) was the only independent predictor of survival. Transplant group. In this cohort, 91 patients (56%) had prolonged QTc (>440 ms) before LT. Mean QTc improved significantly after LT (429 +/- 29 ms vs. 450 +/- 39 ms P<0.002). Of the 91 patients with prolonged QTc, 50 (55%) normalized, three (3.3%) remained unchanged, 12 (13.3%) showed further prolongation, and 26 (28%) showed improvement but remained above normal limits. An additional nine patients who had normal QTc before LT developed prolonged QTc (>440 ms) after LT. CONCLUSION: A prolonged QTc interval was common in patients with cirrhosis, but its presence had no independent effect on mortality. Prolonged QTc returns to normal values in about half of the patients after LT, suggesting that liver disease plays a role, but may not be the only factor in the pathogenesis of prolonged QTc. PMID- 12895264 TI - No association between GSTP1 gene aberrant promoter methylation and prognosis in surgically resected hepatocellular carcinoma patients from the Basque Country (Northern Spain). AB - AIMS: Hepatocellular carcinoma (HCC) is one of the most common cancers worldwide, being linked etiologically to several factors. Glutathione-S-transferases (GSTs) are a family of enzymes that play an important role in detoxification. Hypermethylation of regulatory sequences at glutathione-S-transferase pi class gene (GSTP1) has been found in different human tumor types. In this study, we have studied the methylation status of the GSTP1 promoter region in patients from the Basque Country (Northern Spain) by methylation-specific PCR (MSP). METHODS AND RESULTS: GSTP1 aberrant promoter methylation was present in 24 of 117 (20.5%) tumor samples being associated with late stages of tumor progression. Patients with multiple HCCs showed different patterns of methylation, which could suggest a different clonal origin of multicentric HCC or different degrees of differentiation. No effect on disease-free survival or overall survival was observed in patients with GSTP1 methylated who underwent curative resection. CONCLUSIONS: We can conclude that GSTP1 promoter CpG island methylation appears to be a less common event during hepatocarcinogenesis in European populations than in Asian populations, being associated with late stages of tumor progression. These findings could also be useful to provide new therapeutic strategies through the use of demethylating agents. PMID- 12895265 TI - High rate of long-term virological response after a 1-year course of interferon +/- ribavirin in chronic hepatitis C relapsers. Results of a 191 patients randomized trial. AB - We investigated the long-term efficacy of a 12-month course of interferon (IFN)+ribavirin in chronic hepatitis C relapsers. We randomized 191 relapsers with a 2:1 ratio to receive 3 million units three times a week of interferon alpha (IFN alpha)-2b+ribavirin (1-1.2 g/day) (group A=127 patients) or IFN alpha 2b (group B=60 patients) of same dosage and duration for 1 year. General and hepatitis C data of group A and B patients were similar. The main goal of the study was to determine the rate of sustained virological response evaluated 1 year after treatment. RESULTS: Virological sustained response (SR) was 61% and 12% for groups A and B, respectively (P<0.001). A significant histological improvement was observed in both treatment groups. The Metavir activity score became significantly lower in the IFN+ribavirin group than in the IFN group (P<0/0001). The Metavir fibrosis scores remained unchanged. Also, at the end of the treatment, the virological response was 69% (88/127) for group A and 33% (20/60) for group B (P<0.001). CONCLUSION: One-year retreatment of relapsers with the combination of IFN+ribavirin led to 61% of virological SR and to a significant improvement of histological activity. Therefore, the therapeutic schedule presented here can be considered of particular interest for the retreatment of relapsers. PMID- 12895266 TI - Fibrosis and glycogen stores depletion induced by prolonged biliary obstruction in the rat are ameliorated by metadoxine. AB - BACKGROUND/AIMS: To evaluate liver-beneficial properties of metadoxine, not related with alcohol metabolism, bioactivation of external toxins or antioxidant mechanisms, the chronic bile duct ligation (BDL) model was used and results were compared with colchicine. METHODS: Seven groups (n=6) of male Wistar rats were used. Four groups were BDL and received metadoxine (60 mg/kg/12 h i.p.), colchicine (10 microg/rat/day/p.o.), both or vehicles; three groups were sham appropriate controls. Collagen content was determined by measuring hydroxyproline in liver samples; malondialdehyde (MDA) was used to estimate lipid peroxidation levels; glycogen was determined utilizing the anthrone reagent; gomory's trichromic stains of liver sections were performed. RESULTS: Collagen increased four-fold by BDL, metadoxine, colchicine or both prevented fibrosis partially; MDA levels increased three-fold by BDL and no treatment had any significant effect; glycogen was almost depleted in the cirrhotic group, metadoxine preserved glycogen; bilirubins, and alanine aminotransferase and gamma glutamyltranspeptidase activities increased several-fold in the BDL group, and both drugs prevented these effects partially. The histopathological analysis correlated with biochemical data. CONCLUSIONS: Both compounds showed similar antifibrotic properties; metadoxine was more effective in preserving glycogen. Besides its antioxidant effects and its ability to induce alcohol metabolism, metadoxine possesses important antifibrotic and antinecrotic properties, and maintains energy stores efficiently. PMID- 12895267 TI - Impact of high-dose interferon induction and ribavirin therapy in patients with chronic hepatitis C relapsing after or not responding to interferon monotherapy. AB - BACKGROUND/AIMS: Initial high-dose interferon-alpha induction therapy in combination with ribavirin improves sustained response rates in treatment-naive patients. This prospective, randomized, controlled study tested whether non responders or relapsers to interferon monotherapy also benefit from induction therapy. METHODS: Patients with chronic hepatitis C who had not responded to (n=75) or relapsed (n=80) after previous interferon therapy were randomized to receive three different interferon doses during the first 14 weeks of therapy (A: 10 MU IntronA/day for 2 weeks, followed by 10 MU/2 days for 12 weeks; B:5 MU/d for 14 weeks; C: 5 MU/2 days for 14 weeks) followed in all by 5 MU/2 days for 24 weeks. All patients received 1-1.2 g ribavirin/day throughout the whole study. RESULTS: The rates of viral clearance at any time on treatment were similar in all groups. Sustained response rates were also not different among the groups in interferon nonresponders (A 32%, B 29%, C 31%) and relapsers (A 64%, B 68%, C 71%), respectively, as well as in patients with different genotypes. As expected, sustained response rates were higher in patients with genotype non-1 than in those with genotype 1. CONCLUSION: High-dose induction therapy does not improve the outcome of interferon/ribavirin therapy in interferon nonresponders or relapsers. PMID- 12895268 TI - Glycine modulates cytokine secretion, inhibits hepatic damage and improves survival in a model of endotoxemia in mice. AB - BACKGROUND AND AIM: There is substantial experimental evidence that the amino acid glycine may have a role in protecting tissues against insults such as ischemia, hypoxia and reperfusion. Our aim was to investigate the ability of the amino acid glycine to prevent hepatic damage induced by injection of lipopolysaccharide and d-galactosamine (d-Gal), to modulate pro- and anti inflammatory cytokine levels, and to improve survival. METHODS: Mice were challenged with an intraperitoneal injection of d-Gal (16 mg/mouse) and lipopolysaccharide (LPS, 1 microg/mouse). The intervention group also received an intraperitoneal injection of glycine (150 mg/kg) in two doses: 24 h before and just after LPS challenge. Serum cytokine levels were measured 2 h after challenge, and liver enzymes and histology were determined 16 h after LPS. Separate groups of mice received the same treatment and the survival rate was determined 24 h and ten days after endotoxin administration. In in vitro experiments, cultured mononuclear cells were stimulated by LPS, and TNF-alpha and IL-10 secretion were measured, in the presence or absence of glycine. RESULTS: In the glycine-treated mice, the serum levels of liver enzymes and TNF-alpha, the histologic necroinflammation score and the mortality rate were significantly reduced compared to control mice (P<0.001). Serum IL-10 levels in the glycine treated mice were increased (P<0.01). In vitro studies in cultured lymphocytes isolated from either normal or glycine pretreated mice, demonstrated a significant and dose-dependent inhibition of LPS-induced TNF-alpha secretion and increase in IL-10 response after treatment with glycine (P<0.01). In conclusion, glycine reduces hepatic damage and improves survival rate in this mouse model of endotoxemia. The protective effect of glycine is associated with modulation of TNF-alpha and IL-10 secretion. PMID- 12895269 TI - CC531s colon carcinoma cells induce apoptosis in rat hepatic endothelial cells by the Fas/FasL-mediated pathway. AB - The mechanisms involved in colorectal carcinoma with liver metastasis are not well known. Metastasizing colon carcinoma cells express more FasL than primary colon carcinoma cells and cancer cells induce apoptosis in hepatocytes by the Fas/FasL pathway. Therefore, this study focused on Fas/FasL expression and functionality in rat liver sinusoidal endothelial cells (LSECs) and CC531s colon carcinoma cells in vitro and in vivo. RT-PCR and immunochemistry revealed Fas and FasL in LSECs and CC531s, respectively. Functionality of Fas was assessed in vitro by incubation with human recombinant FasL (1-100 ng/ml) with or without enhancer. At concentrations of 10 and 100 ng/ml with enhancer, respectively 21% and 44% of endothelial cells showed signs of apoptosis using Hoechst 33342/propidium iodide staining and electron microscopy. In co-cultures, apoptosis could be detected in endothelial cells neighboring the CC531s and could be inhibited by an antagonistic FasL antibody. Moreover, 18 h after mesenteric injection of CC531s, the sinusoidal endothelium revealed disruption. In conclusion, (i). CC531s cells induce apoptosis in LSECs in vitro by using Fas/FasL; (ii). CC531s cells damage the sinusoidal endothelial lining in vivo; and (iii). this might provide FasL-positive tumor cells a gateway towards the hepatocytes. PMID- 12895270 TI - Insulin resistance/beta-cell function and serum ferritin level in non-diabetic patients with hepatitis C virus infection. AB - BACKGROUND/AIMS: Since impaired glucose tolerance and iron overload are frequently demonstrated in hepatitis C virus (HCV)-related liver diseases, in this study we investigated insulin resistance, pancreatic beta-cell function, i.e., insulin secretion, and serum ferritin levels in patients with HCV infection, especially non-diabetic patients. METHODS: Homeostasis model assessments for insulin resistance (HOMA-IR) and beta-cell function (HOMA-beta) were performed in 92 HCV-infected patients. RESULTS: The levels of plasma immunoreactive insulin (IRI), HOMA-IR, and HOMA-beta were significantly correlated with fasting plasma glucose (FPG) levels. Among the 86 non-diabetics (with an FPG of <126 mg/dl), IRI, HOMA-IR, and HOMA-beta were significantly higher in patients with liver cirrhosis than in patients with persistently normal alanine aminotransferase levels. The IRI and HOMA-IR values, but not the HOMA beta values, were correlated with both serum transaminase and ferritin levels in the 65 non-diabetic chronic hepatitis patients. CONCLUSION: Insulin resistance was connected with impaired glucose tolerance and the severity of liver diseases in non-diabetic patients with HCV infection. Iron overload may be responsible for insulin resistance, or vice versa. Pancreatic beta-cell function was unrelated to the patients' serum ferritin levels. PMID- 12895271 TI - Antiapoptotic and protective effects of roscovitine on ischemia-reperfusion injury of the rat liver. AB - OBJECTIVE: This study was designed to investigate the role of apoptosis on liver IR injury and to determine the effects of roscovitine on this process. MATERIALS AND METHODS: Rat livers pretreated with roscovitine received 60 min right lobe ischemia followed by 4 h (n=8) and 24 h (n=8) reperfusion. Tissue injury was evaluated by serum alanine aminotransferase, aspartate aminotransferase, tissue malondialdehyde, myeloperoxidase measurements and histological examination. Dead (apoptotic and necrotic) hepatocytes were determined by trypan blue dye and apoptosis was evaluated with M30 monoclonal antibody. RESULTS: Administration of roscovitine significantly decreased both apoptotic and dead hepatocyte counts compared to controls (P<0.01). Liver transaminase levels were decreased significantly in the roscovitine-pretreated groups (P<0.05). In the groups evaluated 24 h after reperfusion, MPO levels (P<0.02) and leukocyte infiltration on histologic sections were decreased significantly in the roscovitine administered group compared to its control. CONCLUSIONS: This study indicates that hepatocyte apoptosis may play a role in the development of IR injury of the liver. Administration of roscovitine may be beneficial in preventing this injury. PMID- 12895272 TI - NMR-derived model of interconverting conformations of an ICAM-1 inhibitory cyclic nonapeptide. AB - We have produced by phage-display a disulfide-linked cyclic nonapeptide (inhibitory peptide-01, IP01), CLLRMRSIC, that binds to intracellular adhesion molecule-1 (ICAM-1) and blocks binding to its counter-structure, leukocyte functional antigen-1 (LFA-1). As a first step towards improving its pharmacologic properties, we have performed a structural and functional analysis of this peptide inhibitor to determine the features relevant to ICAM-1 binding. We report here the solution model of our initial product, IP01, as derived from two dimensional nuclear magnetic resonance (NMR) restraints and molecular modeling. Distance and dihedral angle restraints, generated from nuclear Overhauser effect spectroscopy (NOESY) and one-dimensional-NMR experiments respectively, were used to generate an ensemble of structures using distance geometry and simulated annealing. Molecular dynamic simulations produced three interconverting conformational families consistent with the NMR-derived constraints. We describe these conformations and their mechanism of interconversion. Furthermore, we have measured the IC50 s of a series of inhibitors generated from IP01 through alanine substitution of each residue. These results show that the L2-L3-R4-M5-R6 segment is functionally active, conformationally flexible, and contains a beta-turn involving residues R4-S7, while the C1-C9-I8-S7 segment is less functionally active but adopts a more defined solution conformation, consistent with a scaffolding function. This model will be useful for designing nonpeptide-based organic inhibitors with improved pharmacologic properties. PMID- 12895273 TI - Induction of carbohydrate-specific antibodies in HLA-DR transgenic mice by a synthetic glycopeptide: a potential anti cancer vaccine for human use. AB - Over the last few years, anticancer immunotherapy has emerged as a new exciting area for controlling tumors. In particular, vaccination using synthetic tumor associated antigens (TAA), such as carbohydrate antigens hold promise for generating a specific antitumor response by targeting the immune system to cancer cells. However, development of synthetic vaccines for human use is hampered by the extreme polymorphism of human leukocyte-associated antigens (HLA). In order to stimulate a T-cell dependent anticarbohydrate response, and to bypass the HLA polymorphism of the human population, we designed and synthesized a glycopeptide vaccine containing a cluster of a carbohydrate TAA B-cell epitope (Tn antigen: alpha-GalNAc-Ser) covalently linked to peptides corresponding to the Pan DR 'universal' T-helper epitope (PADRE) and to a cytotoxic T lymphocyte (CTL) epitope from the carcinoembryonic antigen (CEA). The immunogenicity of the construct was evaluated in outbred mice as well as in HLA transgenic mice (HLA DR1, and HLA-DR4). A strong T-cell dependent antibody response specific for the Tn antigen was elicited in both outbred and HLA transgenic mice. The antibodies induced by the glycopeptide construct efficiently recognized a human tumor cell line underlying the biological relevance of the response. The rational design and synthesis of the glycopeptide construct presented herein, together with its efficacy to induce antibodies specific for native tumor carbohydrate antigens, demonstrate the potential of a such synthetic molecule as an anticancer vaccine candidate for human use. PMID- 12895274 TI - Model studies of the function of blockers on the small conductance potassium ion channel. AB - A correlation between KI (equilibrium dissociation constants) and IC50 (concentration at 50% inhibition) inhibitors for the family of blockers of the small conductance potassium ion channels and their intrinsic characteristics like molecular mass and volume have been investigated. Most of the blockers in the family are not selective, in contrast to apamin - an 18 amino acid bee venom toxin - that is known to be a highly potent and selective blocker of these channels. Differences and similarities between the blockers have been analyzed, pointing toward the origin of their selectivity and relative potency. In conclusion, an ion channel blocking is a process controlled mainly by diffusion, in accordance with previous experimental results. PMID- 12895276 TI - Effects of acute tryptophan depletion on executive function in healthy male volunteers. AB - BACKGROUND: Neurocognitive impairment is frequently described in a number of psychiatric disorders and may be a direct consequence of serotonergic dysfunction. As impairments in executive functions are some of the most frequently described, the purpose of this study was to examine the performance of normal volunteers on a range of executive tasks following a transient reduction of central serotonin (5-HT) levels using the method of acute tryptophan depletion (ATD). METHODS: Fifteen healthy male subjects participated in a within-subject, double-blind, counterbalanced crossover study. ATD was induced by ingestion of a 100 g amino-acid drink. Executive function was evaluated using the Wisconsin Card Sorting Test, Stroop, Verbal Fluency and Trail Making. Visual analogue scales were administered to assess mood. RESULTS: Plasma free and total tryptophan concentrations were significantly reduced by the depleting drink (P < 0.001). ATD selectively improved motor speed/ attention on the Trails A test (P = 0.027), with no effect on subjective ratings of mood. Interaction effects between drink and the order of drink administration were observed on most neurocognitive tests. CONCLUSIONS: The improvement in simple motor speed/ attention following ATD is in keeping with the ascribed role of 5-HT in the cortex, however performance on tests of executive function is not robustly altered. The presence of interaction effects on most tasks suggests that subtle changes may occur but are masked, possibly by simple learning effects, in the context of a crossover design. This has implications for the design of future studies, particularly those examining executive functions. PMID- 12895277 TI - Local variation in helminth burdens of bank voles (Clethrionomys glareolus) from ecologically similar sites: temporal stability and relationships with hormone concentrations and social behaviour. AB - Populations of bank voles (Clethrionomys glareolus) in a fragmented forest habitat in north-east Poland showed local differences in helminth infection intensity, morphometric measures and organ weights that were consistent with differences at the same locations two years previously. Although overall intensities of infection were lower than previously, and there were some differences in the relative intensities of individual helminth species, site differences remained significant and were consistent across replicated subsites. In keeping with site differences in helminth infection and adrenal gland weight and asymmetry, voles at site 1 (high intensity infection) had higher circulating concentrations of corticosterone than those at site 2 (low intensity infection). Since males were sampled outside the breeding season, and thus non-scrotal, testosterone levels were low and did not differ between sites. As previously, voles at site 1 also showed greater hind foot asymmetry. Dyadic interactions between males from the same and different sites in the laboratory showed that males from site 1 were significantly less aggressive, especially when confronted with intruder males from site 2. There was no relationship between aggressiveness and intensity of infection overall or at site 1, but a significant negative relationship emerged at site 2. Aggression thus appeared to be downregulated at the higher intensity site independently of individual levels of infection. Terminal corticosterone concentrations were greater at site 1 and lower among residents that initiated more aggression. While corticosterone concentrations rose over the period of testing, they did not correlate with the amount of aggression initiated or received. PMID- 12895278 TI - Local variation in helminth burdens of Egyptian spiny mice (Acomys cahirinus dimidiatus) from ecologically similar sites: relationships with hormone concentrations and social behaviour. AB - Populations of Egyptian spiny mice (Acomys cahirinus dimidiatus) in a fragmented montane wadi system in the Sinai showed significant differences in the abundance of gut helminths. Differences in parasite load between populations were positively associated with measures of androgen activity but showed no significant relationship with glucocorticoid activity. Social discrimination tests with adult males from different wadis showed that those from sites with greater helminth abundance were less likely to investigate odours from other males and were less aggressive when subsequently interacting with the odour donors. Subjects showed markedly more investigation towards the odours of males from distant wadis compared with those from their own or immediately neighbouring wadi, but were less aggressive when confronted with odour donors from distant wadis. Despite this, there was a positive relationship between the amount of investigation towards distant male odour and subsequent aggression towards the male. While aggressiveness was positively associated with measures of androgen and glucocorticoid activity, no significant relationship emerged with individual helminth infection. Thus aggressiveness appeared to relate to overall local population levels of infection rather than individual challenge. PMID- 12895279 TI - Interactions between Schistosoma intercalatum (Zaire strain) and S. mansoni. AB - Schistosoma mansoni and S. intercalatum, two schistosomes from different evolutionary lineages, are parasitic in humans and therefore able to co-infect the same host where they occur sympatrically in Africa. Previous studies of mating interactions between these species in mice, using the Lower Guinea strain of S. intercalatum, have demonstrated the competitive dominance of S. mansoni over S. intercalatum in terms of pairing ability, which is potentially an important mechanism restricting the distribution of S. intercalatum in Africa. The study presented here examines the mating interactions in mice between S. mansoni and the Zaire (Democratic Republic of Congo) strain of S. intercalatum, which differs from the Lower Guinea strain in many biological characteristics. Analysis of the data showed a preponderance of intraspecific pairs over interspecific, demonstrating a specific mate preference system for both species. Mating competition between these species and the ability of males of both species to effect a change of mate by pulling paired females away from their partners was indicated. Comparisons are made between the competitive mating abilities of both strains of S. intercalatum relative to those of S. mansoni, with the data suggesting that S. mansoni is competitively dominant to S. intercalatum (Zaire) in sequential infections but to a lesser extent than for S. intercalatum (Lower Guinea). Additional factors which may contribute to the confinement of S. intercalatum (Zaire) to the Democratic Republic of Congo are discussed. PMID- 12895280 TI - Modulation of liver fibrosis and pathophysiological changes in mice infected with Mesocestoides corti (M. vogae) after administration of glucan and liposomized glucan in combination with vitamin C. AB - The effects of glucan and liposomized glucan, alone or co-administered with vitamin C, and empty liposomes on hepatic fibrosis in mice infected with Mesocestoides corti (M. vogae) tetrathyridia were studied. Preparations were administered every third day from day 7 to day 31 post-infection (p.i.), nine doses in total. Activities of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and cholesterol levels were measured in sera collected on days 11, 15, 21, 28, 32, 42, 50 and 65 p.i. Liver fibrosis was studied on the same days by measuring hydroxyproline concentration, which is considered a marker for collagen content. Larvicidal effects of the glucan and liposome preparations were estimated on day 65 p.i. in the liver and peritoneal cavity. Glucan formulations significantly enhanced collagen content, most prominently after administration of liposomized glucan in combination with vitamin C. Activities of both enzymes and cholesterol levels were slightly modified after administration of glucan alone. Liposomized glucan with vitamin C significantly increased ALT and AST activity and cholesterol levels up to days 28-32 p.i., after which they plateaued or declined. The most pronounced decrease was after administration of liposomized glucan and vitamin C. The same pattern of biochemical parameters in serum was observed after administration of empty liposomes, however, collagen content was not modified significantly. Larval counts in the liver and the peritoneal cavity were significantly reduced after treatment with either glucan formulation, but were unaffected following treatment with empty liposomes. In summary, intense fibrosis in the liver of mice treated with liposomized glucan and vitamin C did not result in the most extensive parenchymal cell injury but, rather in the highest efficacy of treatment. Liposomal lipids were probably utilized in the reparation of the damaged parenchymal cells, while glucan stimulated phagocytic cells. PMID- 12895281 TI - Embryogenesis and the first-stage larva of Thelazia lacrymalis. AB - The female reproductive system of Thelazia lacrymalis (Nematoda: Thelaziidae) was investigated by light and scanning electron microscopy (SEM) with regard to the developmental stages and the stage deposited by the gravid nematode. Female T. lacrymalis have a didelphic and opisthodelphic type of reproductive system with paired ovaries, oviducts and uteri and a single vagina and vulva. Round and spindle-shaped primary oocytes are documented within the ovaries and oviducts, respectively. The distal part of each uterus provides a fertilization chamber filled with spermatozoa, followed by a sphincter-like part. Further anterior, the uteri broaden gradually containing dividing zygotes, small and large morulae, tadpole-stage embryos and horseshoe-shaped embryos which increase in length and become slimmer forming pretzel-stage embryos and larvae rolled up. The larvae stretch gradually and finally lie straight but still covered with their egg membrane in the vagina. The egg membrane encloses the whole larva and is enlarged at the pointed tail of the larva forming a bulb. At the SEM level, the first stage larva is shown to have a terminal mouth and three hooks directed posteriorly and a striated cuticle. As morphologically identical larvae were also found in lavages of the conjunctival sac of horses infected with T. lacrymalis, this nematode species can be described as ovoviviparous. PMID- 12895282 TI - Effects of temperature on survival, infectivity and in vitro encystment of the cercariae of Echinostoma caproni. AB - The effects of temperature on survival, infectivity and in vitro encystment of Echinostoma caproni cercariae in artificial spring water (ASW) were studied. Effects of aging cercariae in ASW at various temperatures showed that at 23 degrees C cercariae achieved 50% survival in 24 h, compared to 92 h at 12 degrees C. Cercariae aged in ASW at 28 and 37.5 degrees C showed 50% survival at 16 and 10 h, respectively. Cercariae aged at different temperatures for various times were used to infect juvenile Helisoma trivolvis (Colorado strain) snails maintained in ASW at 23 degrees C. Index of infectivity was based on counting encysted metacercariae in the snails at 8 to 12 h post-infection. Cercariae aged at 23, 28 and 37.5 degrees C showed 50% encystment at 6, 8 and 4 h, respectively. Cercariae aged at 4 degrees C showed 50% encystment in 10 h and cercariae aged at 12 degrees C showed 50% encystment beyond 16 h. Cercariae showed maximal longevity and infectivity in snails when aged at 12 degrees C in ASW. For E. caproni, as in other digeneans, the infective period of cercariae is markedly shorter than the maximal life-span at any given temperature. Studies on in vitro encystment of E. caproni cercariae in Locke's solution:ASW (1:1) showed that encystment was optimal at 23 degrees C (78% encystment) and that it declined to 44% at 28 degrees C and became almost nil (0.02%) at 12 or 37.5 degrees C. PMID- 12895283 TI - Identities of two Paragonimus species from Sri Lanka inferred from molecular sequences. AB - Metacercariae of Paragonimus spp. were obtained from field-collected freshwater crabs in Sri Lanka. Genomic DNA was extracted from single metacercariae. Two gene regions (partial mitochondrial cytochrome c oxidase subunit 1 (CO1) and the second internal transcribed spacer of the nuclear ribosomal gene repeat (ITS2)) were amplified using the polymerase chain reaction. Two differing sequences were obtained for each of these gene regions. Phylogenetic analyses placed the type 1 sequences as sister to a clade containing P. westermani and P. siamensis whereas the type 2 sequences were close to published sequences of P. siamensis from Thailand. The possible taxonomic status of these two types are discussed. This is the first report of molecular data about Paragonimus from Sri Lanka. PMID- 12895284 TI - Syphacia obvelata infections and reproduction of male domestic mice Mus musculus domesticus on a sub-Antarctic Island. AB - The reproductive activity of feral male mice on an island of the sub-Antarctic Kerguelen archipelago was influenced by biological factors depending on periods within the breeding season. After having controlled host reproductive activity indices for body size, i.e. age, and body condition effects, Syphacia obvelata prevalence did not vary with host reproductive status or age either during the beginning or the middle-end of the reproductive season. Considering the beginning of the breeding season, worm abundance was more pronounced in males the year following a strong winter crash of the population than in years when high over wintering survival occurred. During the middle-end of the breeding season, males with the highest reproductive status were more infected than males with a lower reproductive status in years when oldest individuals dominated the population. It is suggested that this situation was due to an endocrine related increased host susceptibility partly influenced by a change in the age structure of the population, and that an increase in worm transmission was not directly related to male activity concurrent with reproductive status, nor to population density. PMID- 12895285 TI - Host specificity and geographical distribution of Eubothrium in European salmonid fish. AB - The host specificity and distribution of Eubothrium crassum (Bloch, 1779) and Eubothrium salvelini (Schrank, 1790), morphologically fairly similar pseudophyllidean tapeworms parasitizing salmonid fish, were critically assessed on the basis of morphological and genetic evaluation of extensive material collected from different definitive hosts and geographical regions in Europe. Eubothrium crassum occurs in fish of the genera Salmo, i.e. salmon (S. salar - both freshwater and marine), sea trout (S. trutta trutta), brown trout (S. trutta fario), and lake trout (S. trutta lacustris), and also in Danubian salmon (Hucho hucho) and vendace (Coregonus albula). Eubothrium salvelini parasitizes Arctic char (Salvelinus alpinus) and brook trout (Salvelinus fontinalis) in Europe, and also whitefish (Coregonus wartmanni). Rainbow trout (Oncorhynchus mykiss), which is not a native European fish species, was found to be a suitable definitive host for both Eubothrium species, which may occur simultaneously in the same fish. Previous records of E. crassum in Arctic char and brook trout, and those of E. salvelini in fish of the genus Salmo were most probably misidentifications. Most studies of Eubothrium have involved salmonids from the northern part of Europe, with few records from southern and south-eastern Europe. This study also confirmed the reliability of the morphology of the apical disc for the discrimination of E. crassum and E. salvelini. PMID- 12895286 TI - Helminth parasites of the wild boar, Sus scrofa, in Luristan province, western Iran and their public health significance. AB - Seven helminth species were obtained from 12 wild boars (Sus scrofa) during a survey from 2000 to 2001 in Luristan province, western Iran. These species include the cestode larvae Cysticercus tenuicollis (25%), C. cellulosae (8.3%), the nematodes Metastrongylus apri (41.6%), M. pudendotectus (16.6%), M. salmi (8.3%), Trichuris suis (8.3%) and the acanthocephalan Macracanthorhynchus hirudinaceus (41.6%). No trematodes were found. Seven wild boars (58.3%) were identified as having at least one helminth species. A single infection was detected in 16.6% of cases, but a three species infection covered the highest rank (25%). All these helminths have been reported from other areas of Iran including the north, northeast and southwest, but not in Luristan. Among seven helminths identified, at least three species are transmissible to humans. The public health significance of these helminths is discussed. PMID- 12895287 TI - Isoenzyme analysis of Schistosoma haematobium, S. intercalatum and their hybrids and occurrences of natural hybridization in Cameroon. AB - Isoelectric focusing of glucose-6-phosphate dehydrogenase (G6PD) produced clearly identifiable profiles for S. haematobium and S. intercalatum and their hybrids. To provide a more detailed analysis of the interactions of S. haematobium and S. intercalatum in South West Cameroon over the last 12 years, G6PD analyses were carried out on individual schistosomes collected in Kumba in 1990, Loum in 1990, 1999 and 2000 and Barombi Mbo and Barombi Kotto in 1999. Studies were also carried out on the two parental species S. haematobium Barombi Mbo, S. intercalatum Edea and subsequent generations of hybrids resulting from laboratory crosses of the two parental species. The isoenzyme analysis demonstrated that the 1990 isolate from Kumba, was a recombinant of S. intercalatum x S. haematobium, and that 30% of individual schistosomes collected in 1990 in Loum were also recombinants. The remainder gave data indicative of S. haematobium. In 1999, 12.5% of individuals from Loum showed recombination and 10% in 2000. Results from the most recent parasitological survey in October 2000 showed the persistence of the recombinant population in addition to that of S. haematobium. There was also evidence of recombination having taken place in Barombi Kotto but not Barombi Mbo. This study demonstrates how the situation has changed over the last 12 years, and emphasizes the importance of assessing morphological, biological and molecular data together to gain a true picture of the rapidly evolving situation. PMID- 12895288 TI - The experimental production of Fasciola hepatica metacercariae from three aquatic populations of Galba truncatula. AB - Laboratory investigations on three aquatic populations of Galba truncatula, originating from the Peruvian Altiplano and French Massif Central, were carried out during three successive snail generations to determine if these populations might be successfully used for the metacercarial production of Fasciola hepatica under experimental conditions. High numbers of surviving snails at day 30 post exposure (>70%), high prevalences of F. hepatica infections (>60%), and prolonged productions of cercariae for a mean period of 35 to 47 days were observed in the three populations, whatever the snail generation. In the Peruvian population, metacercariae of F. hepatica significantly decreased in numbers from a mean of 251 in the parent snails to 124 per snail in the F2 generation, whereas no significant variation was observed in the two French populations. As these aquatic snails rarely emerged out of water, the use of these populations for the commercial production of F. hepatica metacercariae was of great interest, because the daily time spent watching the breeding boxes of snails was clearly shorter, thereby reducing the cost of producing metacercariae compared with using amphibious snails reared with romaine lettuce. PMID- 12895289 TI - Ethanol-eluted extract of Rhus verniciflua stokes inhibits cell growth and induces apoptosis in human lymphoma cells. AB - Rhus verniciflua Stokes (RVS) has been used as a traditional herbal medicine. Several earlier studies indicated that an ethanol extract of RVS has both anti oxidant and anti-tumor properties, although the mechanism for the activity remains to be elucidated. In this report, we prepared a highly purified ethanol extract from RVS, named REEE-1 (Rhus ethanol eluted extract-1), and investigated the mechanism involved in its growthinhibitory effect on the human B and T lymphoma cell lines, BJAB and Jurkat, respectively. Results from tritium uptake proliferation assays showed that the proliferative capacities of both BJAB and Jurkat cells were strongly suppressed in the presence of REEE-1. This was further confirmed through trypan blue exclusion experiments that revealed a dose dependent decrease in viable cell numbers after REEE-1 treatment. REEE-1-mediated suppression of cell growth was verified to be apoptotic, based on the increase in DNA fragmentation, low fluorescence intensity in nuclei after propidium iodide staining, and the appearance of DNA laddering. In particular, REEE-1 exerted its anti-oxidant activity through the inhibition of hydroxyl radical-mediated degradation by iron ion chelation rather than direct scavenging of hydroxyl radicals. Furthermore, REEE-1 was revealed to be a potential scavenger of superoxide anions. Collectively, our findings suggest that REEE-1 is a natural anti-oxidant that could be used as a cancer chemo-preventive and therapeutic agent. PMID- 12895290 TI - Identification of mutations in protein kinase CKIIbeta subunit that affect its binding to ribosomal protein L41 and homodimerization. AB - Protein kinase CKII is composed of two catalytic (alpha or alpha') subunits and two regulatory (beta) subunits. The CKIIbeta subunit is thought to mediate the tetramer formation and interact with other target proteins. However, its physiological function remains obscure. In this study, point mutants of CKIIbeta that are defective for the L41 binding were isolated by using the reverse two hybrid system. A sequence analysis of the point mutants revealed that Asp-26, Met 52, and Met-78 of CKIIbeta are critical for L41 binding; Asn-67 (and/or Lys-139) and Met-52 are important for CKIIbeta homodimerization. Two point mutants, R75 and R83, of CKIIbeta interacted with L5, topoisomerase IIbeta, and CKBBP1/SAG, but not with the wild-type CKIIbeta. This indicates that CKIIbeta homodimerization is not a prerequisite for its binding to target proteins. These CKIIbeta point mutants may be useful in exploring the biochemical physiological functions of CKIIbeta. PMID- 12895291 TI - Possibility of using DNA chip technology for diagnosis of human papillomavirus. AB - To explore the application of DNA chip technology for the detection and typing of Human Papillomavirus (HPV), the HPV6, 11, 16 and 18 gene fragments were isolated and printed onto aminosilane-coated glass slides by a PixSys 5500 microarrayer as probes to prepare the HPV gene chips. HPV samples, after being labeled with fluorescent dye by restriction display PCR (RD-PCR) technology, were hybridized with the microarray, which was followed by scanning and analysis. The experimental condition for preparing the HPV gene chips was investigated, and the possibility of HPV genotyping using gene chips was discussed. The technique that was established in this study for preparing HPV gene chips is practical. The results of the present study demonstrated the versatility and inspiring prospect of using this technology to detect and genotype HPV. PMID- 12895293 TI - Effects of zinc on the activity and conformational changes of arginine kinase and its intermediate. AB - The effects of zinc on arginine kinase and its collapsed-state intermediate were studied. Both arginine kinase and the collapsed-state intermediate were inactivated in the presence of zinc, following a biphasic kinetic course. The corresponding apparent rate constants of inactivation at different zinc concentrations and conformational changes in the presence of 0.5 mM zinc were obtained. The conformational changes of arginine kinase and the collapsed-state intermediate were followed by fluorescence spectra and circular dichroism spectra. Comparison of the results for arginine kinase and the collapsed-state intermediate showed that the collapsed-state intermediate was more susceptible to zinc, which indicated that the collapsed-state intermediate was more flexible and unstable than arginine kinase. The special structure of arginine kinase might explain these diverse phenomena. PMID- 12895292 TI - Translocation and phosphorylation of calcyclin binding protein during retinoic acid-induced neuronal differentiation of neuroblastoma SH-SY5Y cells. AB - For better understanding of functions of the Calcyclin Binding Protein (CacyBP) and exploring its possible roles in neuronal differentiation, the subcellular localization of human CacyBP was examined in retinoic acid(RA)-induced and uninduced neuroblastoma SH-SY5Y cells. Immunostaining indicated that CacyBP was present in the cytoplasm of uninduced SH-SY5Y cells, in which the resting Ca(2+) concentration was relatively lower than that of RA-induced cells. After the RA induction, immunostaining was seen in both the nucleus and cytoplasm. In the RA induced differentiated SH-SY5Y cells, CacyBP was phosphorylated on serine residue(s), while it existed in a dephosphorylated form in normal (uninduced) cells. Thus, the phosphorylation of CacyBP occurs when it is translocated to the nuclear region. The translocation of CacyBP during the RA-induced differentiation of SH-SY5Y cells suggested that this protein might play a role in neuronal differentiation. PMID- 12895294 TI - Kinetics of denaturation of human and chicken hemoglobins in the presence of co solvents. AB - The stability of four hemoglobins (Hb) in dimer forms (low concentration) were investigated by the kinetics of denaturation. The rate constants of denaturation were obtained by variation of 280 nm absorption versus time in 10 mM Tris-HCl, 10 mM EDTA, pH 8.0 at 45 degrees C in the absence and presence of 0.5 M ethanol, dimethyl sulfoxide (DMSO), formamide, and glycerol. The results show the trend of rate constants in different co-solvents in the following order: chicken hemolysate < human hemolysate and chicken Hb D < chicken Hb A. The buried surface area was calculated for Hb samples in the absence of co-solvents. Accordingly, the trend points out that: chicken Hb D > chicken Hb A > human Hb A. These results suggest that both chicken hemolysate and chicken Hb D are relatively more stable than human and chicken Hb A, respectively. However, the denaturation rate constants of Hb in different co-solvents have designated the following order: ethanol > DMSO > formamide > glycerol. As a matter of fact, this phenomenon is an indication of an increase in the denaturation capacity (DC) and hydrophobicity, and a decrease in the surface tension of the solution in the preceding co solvents. PMID- 12895295 TI - L-arginine ameliorates kidney function and urinary bladder sensitivity in experimentally-induced renal dysfunction in rats. AB - Effects of L-arginine and NG-nitro-L-arginine methyl ester (L-NAME) on the renal dysfunction that is induced by cisplatin (CDDP) were investigated. A single dose of CDDP (7.5 mg/kg i.p.) induced renotoxicity, which was manifested by increasing the sensitivity of isolated urinary bladder rings to acetylcholine (ACh), together with a significant elevation of serum urea and creatinine, and a severe decrease in serum albumin. Moreover, renal dysfunction was further confirmed by a significant decrease of enzyme activities, such as glutathione peroxidase, GSH-Px (E.C 1.11.1.9), catalase (E.C 1.11.1.6), as well as a significant increase in lipid peroxides that were measured as malondialdhyde (MDA) in kidney tissue homogenates. The administration of L-arginine (70 mg/kg/d p.o in drinking water 5 d before and 5 d after the CDDP injection) significantly ameliorated the renotoxic effects of CDDP, as judged by restoring the normal responses of isolated bladder rings to Ach, and also by an improvement in a range of renal function indices, which included serum urea and creatinine concentrations and kidney weight. In addition, L-arginine prevents the rise of MDA, as well as a reduction of GSH-Px and catalase activities in kidney tissues homogenates. On the other hand, the administration of L-NAME (4 mg/kg/d p.o) resulted in no protection against renal dysfunction that was induced by CDDP treatment. The findings of this study suggest that L-arginine can attenuate kidney injury that is produced by CDDP treatment. In addition, L-arginine may be a beneficial remedy for CDDP-induced renal toxicity, and could be used to improve the therapeutic index of CDDP. PMID- 12895296 TI - Gene therapy for mice sarcoma with oncolytic herpes simplex virus-1 lacking the apoptosis-inhibiting gene, icp34.5. AB - A mutant herpes simplex virus 1, mtHSV, was constructed by inserting the E. coli beta-galactosidase gene into the loci of icp34.5, the apoptosis-inhibiting gene of HSV. The mtHSV replicated in and lysed U251 (human glioma cells), EJ (human bladder cells), and S-180 (mice sarcoma cells), but not Wish (human amnion cells) cells. With its intact tk (thymidine kinase) gene, mtHSV exhibited susceptibility to acyclovir (ACV), which provided an approach to control viral replication. An in vivo test with mtHSV was conducted in immune-competent mice bearing sarcoma S 180 tumors, which were treated with a single intratumoral injection of mtHSV or PBS. Tumor dimensions then were measured at serial time points, and the tumor volumes were calculated. Sarcoma growth was significantly inhibited with prolonged time and reduced tumor volume. There was microscopic evidence of necrosis of tumors in treated mice, whereas no damage was found in other organs. Immunohistochemical staining revealed that virus replication was exclusively confined to the treated tumor cells. HSV-1 DNA was detected in tumors, but not in the other organs by a polymerase chain reaction analysis. From these experiments, we concluded that mtHSV should be a safe and promising oncolytic agent for cancer treatment. PMID- 12895297 TI - The effect of willow leaf extracts on human leukemic cells in vitro. AB - The young developing leaves of willow (Salix safsaf, Salicaceae) trees have antileukemic activity. After a 24-h incubation in vitro, the crude water extracts of the leaves killed a majority of the blasts of acute myeloid leukemia (AML, 73.8%). PMID- 12895298 TI - Development of an in vitro assay for the proteolytic processing of the CDP/Cux transcription factor. AB - The CDP/Cux transcription factor was previously shown to be proteolytically processed at the G1/S transition. In view of characterizing and eventually identifying the protease responsible for CDP/Cux processing, we have established an in vitro proteolytic processing assay. CDP/Cux recombinant proteins expressed in mammalian or bacterial cells were efficiently processed in vitro using as a source of protease either whole cell extracts, the nuclear or the cytoplasmic fraction. Processing was found to take place optimally at a lower pH, to be insensitive to variations in salt concentration, and to be inhibited by the protease inhibitors MG132 and E64D. Interestingly, the bacterially-produced substrate was more efficiently processed than the substrate purified from mammalian cells. Moreover, processing in vitro was more efficient when CDP/Cux substrates were purified from populations of cells enriched in the S phase than in the G1 phase of the cell cycle. Altogether, these results suggest that posttranslational modifications of CDP/Cux in mammalian cells inhibits processing and contributes to the cell cycle-dependent regulation of processing. The in vitro processing assay described in this study will provide a useful tool for the purification and identification of the protease responsible for the processing of CDP/Cux. PMID- 12895299 TI - Ras oncogene mutations in urine sediments of patients with bladder cancer. AB - Early detection of bladder cancer is particularly important since it dramatically affects the survival rates. However, neither urinary cytology nor tumor markers that are currently used are sensitive enough for the early detection of bladder cancer or recurrent disease. The ras genes are frequently mutated in cancer. In this study, we investigated the diagnostic potential of ras mutation analysis in urinary sediments of patients with bladder cancer using a single-strand conformation polymorphism analysis and polymerase chain reaction. Mutation in codon 12 of the H-ras gene was observed in 39% of the patients. Our results indicate that this approach may significantly improve diagnostic sensitivity in detecting bladder tumors. PMID- 12895300 TI - Effects of neutral, cationic, and anionic chromium ascorbate complexes on isolated human mitochondrial and genomic DNA. AB - The relative activities of neutral, cationic, and anionic chromium ascorbate complexes toward isolated human mitochondrial and genomic DNA were investigated at physiologically relevant conditions by agarose gel electrophoresis. A direct relationship between the charge of the Cr(III) species and their DNA-damaging properties was found. The cationic species were found to be fully capable of DNA cleavage, even in short incubation periods. Incubations were also performed in the presence of amino acids. No apparent effect was observed under the applied experimental conditions to facilitate or prevent damage through the ternary amino acid-Cr-DNA adduct formation or binary chromium-amino acid complex formation. PMID- 12895301 TI - Identification of essential histidines in cyclodextrin glycosyltransferase isoform 1 from Paenibacillus sp. A11. AB - The isoform 1 of cyclodextrin glycosyltransferase (CGTase, EC 2.4.1.19) from Paenibacillus sp. A11 was purified by a preparative gel electrophoresis. The importance of histidine, tryptophan, tyrosine, and carboxylic amino acids for isoform 1 activity is suggested by the modification of the isoform 1 with various group-specific reagents. Activity loss, when incubated with diethylpyrocarbonate (DEP), a histidine modifying reagent, could be protected by adding 25 mM methyl beta-cyclodextrin substrate prior to the modification. Inactivation kinetics of isoform 1 with DEP resulted in second-order rate constants (k(inactivation)) of 29.5 M(-1)s(-1). The specificity of the DEP-modified reaction for the histidine residue was shown by the correlation between the loss of isoform activity and the increase in the absorbance at 246 nm of N-carbethoxyhistidine. The number of histidines that were modified by DEP in the absence and presence of a protective substrate was estimated from the increase in the absorbance using a specific extinction coefficient of N-carbethoxyhistidine of 3,200 M(-1)cm(-1). It was discovered that methyl-beta-CD protected per mole of isoform 1, two histidine residues from the modification by DEP. To localize essential histidines, the native, the DEP-modified, and the protected forms of isoform 1 were digested by trypsin. The resulting peptides were separated by HPLC. The peptides of interest were those with R(t) 11.34 and 40.93 min. The molecular masses of the two peptides were 5,732 and 2,540 daltons, respectively. When the data from the peptide analysis were checked with the sequence of CGTase, then His-140 and His 327 were identified as essential histidines in the active site of isoform 1. PMID- 12895302 TI - A colorimetric microplate assay method for high throughput analysis of lipase activity. AB - The present work describes a colorimetric microplate assay for lipase activity based on the reaction between 5,5'-dithiobis(2-nitro benzoic acid) (DTNB) and the hydrolysis product of 2,3-dimercapto-1-propanol tributyrate (DMPTB). Reaction mixtures containing DTNB, DMPTB, and lipase were prepared in microplate wells, and the absorbance at 405nm was recorded after incubation at 37 degrees C for 30 min. A linear relationship was obtained in the range of 0.1-1 U of lipase activity by this method. The reaction conditions were also optimized for the range of 0.01-0.1 U or 1-10 U. When assaying crude tissue extracts, the reaction of DTNB with non-specific reducing agents created a major source of error. However, this error was corrected by the use of blank samples that did not contain DMPTB. PMID- 12895303 TI - A generic time-resolved fluorescence assay for serine/threonine kinase activity: application to Cdc7/Dbf4. AB - The serine/threonine protein kinase family is a large and diverse group of enzymes that are involved in the regulation of multiple cellular pathways. Elevated kinase activity has been implicated in many diseases and frequently targeted for the development of pharmacological inhibitors. Therefore, non radioactive antibody-based kinase assays that allow high throughput screening of compound libraries have been developed. However, they require a generation of antibodies against the phosphorylated form of a specific substrate. We report here a time-resolved fluorescence assay platform that utilizes a commercially available generic anti-phospho-threonine antibody and permits assaying kinases that are able to phosporylate threonin residues on protein substrates. Using this approach, we developed an assay for Cdc7/Dbf4 kinase activity, determined the K(m) for ATP, and identified rottlerin as a non-ATP competitive inhibitor of this enzyme. PMID- 12895304 TI - [Analysis of maternal mortality in China from 1996 to 2000]. AB - OBJECTIVE: To study the trend and characteristics of maternal mortality in China from 1996 to 2000. METHODS: Population-based epidemiological survey in 116 monitoring units in China were used. RESULTS: Maternal mortality in China dropped by 17.1% from 63.9 per 100,000 live births in 1996 to 53.0 in 2000, in rural area dropped by 22.2% from 86.4 per 100,000 live births to 67.2, and in urban area only 1.0% from 29.2 per 100 000 live births to 28.9, the leading causes of maternal mortality in China are hemorrhage, preeolampsia/eclampsia and amniotic fluid embolism the maternal mortality due to hemorrhage in national level and rural level has declined by 33.8%, 34.9% respectively. CONCLUSION: The maternal mortality appears a declined trend on both the national and rural levels in China from 1996 to 2000, the maternal mortality due to hemorrhage dose also, bur the maternal mortality in urban area keeps unstable. PMID- 12895305 TI - [Maternal plasma leptin levels and their relationship to insulin and glucose in pregnant women with gestational diabetes mellitus and gestational impaired glucose tolerance]. AB - OBJECTIVE: To investigate the changes in leptin levels and the relationship between the substance and insulin and glucose in pregnant women with gestational diabetes mellitus (GDM) and gestational impaired glucose tolerance (GIGT). METHODS: Radio immunoassay was used to measure the leptin levels at fasting and 3 h after oral glucose load (50 g) in 36 patients with GDM and GIGT, and also in 24 normal pregnant women. In each pregnant women, fasting serum insulin levels and glycosylated haemoglobin values were measured by electrochemiluminescent immunoassays and pressure liquid chromatography (LPLC) respectively, glucose levels 1 h after the administration of glucose by glucose oxidase method. RESULTS: The serum leptin levels in pregnant women with GDM and GIGT and normal pregnant women were (14.9 +/- 4.3) micro g/L and (10.0 +/- 1.8) micro g/L, respectively, suggesting the serum leptin levels were significantly higher in pregnant women with GDM and GIGT than in normal controls. The levels of fasting serum insulin, glycosylated haemoglobin values and plasma glucose levels obtained 1h after oral administration of 50 g glucose were (12.9 +/- 4.3) mU/L, (6.1 +/- 1.1)%, (11.0 +/- 1.4) mmol/L, respectively, whereas the corresponding values in normal controls were (8.6 +/- 3.2) mU/L, (4.5 +/- 1.0)%, (7.8 +/- 1.2) mmol/L. A positive correlation was found between maternal serum leptin levels and fasting serum insulin levels in pregnant women with GDM and GIGT (r = 0.835, P < 0.01). A positive correlation was also found between maternal leptin concentrations and glycosylated haemoglobin values, as well as between leptin levels and plasma glucose levels obtained 1 h after the administration of 50g glucose in women with GDM and GIGT (r = 0.758, P < 0.01; r = 0.561, P < 0.01). CONCLUSIONS: Leptin levels are elevated in pregnant women with GDM and GIGT compared with healthy pregnant women, a positive and significant correlation was found between the maternal leptin levels and fasting insulin levels, as well as between the maternal leptin levels and plasma glucose levels obtained 1 h after the administration of 50 g glucose in women with GDM and GIGT. PMID- 12895306 TI - [Study of the cervix of normal pregnancy and threatened preterm delivery using transvaginal sonography]. AB - OBJECTIVE: To observe the changes of cervix in women between 16 and 35 weeks of gestation using transvaginal ultrasound and to determine the value of the cervix length in predicting preterm delivery among the women with threatened preterm labor. METHODS: Transvaginal sonography were performed in 154 normal single nulliparous pregnant women between 16 and 35 weeks of gestation and 58 women with threatened preterm labor but without premature rupture of membrane. RESULTS: (1) There is no definitely change in either cervical length or internal diameter of cervix among the pregnant women over the studied period. There was also no funneling to be observed. The mean cervical length is (36 +/- 5) mm, and the mean cervical internal diameter is (4 +/- 1) mm. (2) Eleven preterm deliveries occurred among the 58 pregnancy women with threatened preterm labor. The cervical length by sonography of 11 preterm deliveries was (18 +/- 6) mm, and was significant shorter than the women without preterm deliveries whose mean cervical length was (32 +/- 6) mm (P < 0.001). The cervical length of women who delivered pretermly was totally 0.05). (3) In Ward triangle, subjects with ER PP genotype had significantly lower BMD than those in ER Pp and pp genotypes (P < 0.05). (4) Women with BbPP genotype combination had lower BMD levels at lumber and hip, and with bbPP and Bbpp genotypes combination significantly higher lumber BMD levels than BbPP genotype (P < 0.05). However, the differences of BMD among subjects with different VDR and ER genotypes became not significant after adjusting the confounder of body weight. CONCLUSIONS: (1) Body weight and BMI play important roles to PBMD of Beijing women. (2) There was no significant difference of BMD levels between VDR genotypes at any site. (3) PvuII polymorphism of ER gene was associated with low Ward triangle BMD. (4) There was significant relationship between the combination of ER and VDR polymorphisms at lumbar and hip BMD. Our data suggest that genetic variation at the ER locus, singly and in relation to the VDR locus, may influence the attainment and maintenance of peak bone mass in young women. PMID- 12895310 TI - [Clinical study about diagnosis and management on 10 women with rectovaginal endometriosis]. AB - OBJECTIVE: To study the diagnosis and therapy of the rectovaginal endometriosis. METHODS: Descriptive retrospective study, between 1992 and 2002, 10 women with rectovaginal endometriosis were treated at the Peking Union Medical College Hospital. RESULTS: The average age were 40 years old. Six patients presented with rectal irritative symptom. Three of them complained about deep dyspareunia. Eight patients complained about mild to moderate dysmenorrhea. The recto-vaginal septum should be carefully palpated. None of these patients had either clinical or objective evidence of ovarian endometriosis, nor was there evidence of any obstructive lesions of the intestine or ureters. The CA(125) levels had a sensitivity of 20%. Ultrasonography failed to find the rectovaginal endometriotic nodules. Diagnosis of the rectovaginal endometriosis was based on symptoms, vaginal and rectal examination. The Revised American Fertility Society (rAFS) stages can not evaluate the grade of the endometriosis. Four patients were given leuprolide acetate depot at 3.75 mg, 1 ampule every 28 days, and treatment had a planned duration of 3 mouths. Follow-up evaluations were set 3 mouths later. The patients showed an improvement with respect to pain during the 3 mouths treatment course but had early pain recurrence after drug suspension. The endometriotic lesions showed a slight but reduction in size during therapy but had returned to original volume within 3 mouths after cessation of the gonadotropin-releasing hormone analog (GnRH-a) treatment. Finally, all patients required further treatment with a surgical solution. Six patients succeeded to remove the nodules by laparotomy. Four patients were failed to remove the total lesions by laparoscope and transvaginal section. CONCLUSIONS: The vaginal and rectal examination should be performed if the patients complained about tectal irritative symptom or dyspareunia. The rAFS stages had no strong association with the severity of the rectovaginal endometriosis. GnRH-a should not be considered a real therapeutic alternative to surgical treatment for patients with symptom endometriosis of rectovaginal septum. Laparotomy is efficient for the treatment of the rectovaginal endometriosis. PMID- 12895311 TI - [Analysis of 16 cases of uterine perforation during hysteroscopic electro surgeries]. AB - OBJECTIVE: To analyse the cause, diagnosis, treatment and preventive methods of uterine perforation resulting from hysteroscopic electro-surgeries. METHODS: Data of cases with uterine perforation were collected from 5 hospitals where overall 3,541 hysteroscopic electro-surgeries were done from May 1990 to July 2002. There were 1 468 transcervical resections of endometrium (TCRE), 797 cases of transcervical resection of myoma (TCRM), 783 cases of transcervical resection of endometrial polyp (TCRP), 189 cases of transcervical resection of uterine septa (TCRS), 112 cases of transcervical resection of uterine adhesion (TCRA) and 192 cases of transcervical removal of foreign body (TCRF). All operations were performed under B-ultrasonographic or laparoscopic monitoring. Cervical dilator stick was inserted into cervical canal or 200 micro g of misoprostol put in the posterior fornix the evening before operation. The procedures were done according to different indications and purposes. Cases of uterine perforation were divided into two groups: caused by approaching (entry-related) and by surgical instruments (technique-related). RESULTS: Totally sixteen cases (0.45%) of uterine perforation occurred. Seven cases occurred during cervical dilatation and 1 during hysteroscopy inserting lentry-related. Eight cases were technique related caused by electrode. The incidences of uterine perforation of different operations were: TCRA 4.46% (5/112), TCRF 3.12% (6/192), TCRE 0.27% (4/1 468), TCRM 0.13% (1/797). TCRP and TCRS none. These 16 cases were all diagnosed during operations. 10 cases (62%) by B ultrasound and (or) laparoscopy, 6 cases (38%) by hysteroscopy and clinical features. 13 cases were complete uterine perforations, among them 2 were diagnosed by laparoscopic monitoring, 5 by B-ultrasonic monitoring, 4 by hysteroscopy and 2 by symptoms and B-ultrasound, 3 cases were incomplete uterine perforations in which 2 were diagnosed by laparoscopic monitoring and one by B-ultrasound monitoring. CONCLUSIONS: Half of uterine perforation cases were entry-related, so attention has to be paid to entry of Hegar or hysteroscopy (i.e., not dilate the cervix as possible and introduce the scope under direct vision). The other half were related to surgeons' experience and type of operation. TCRA and TCRF run more risks. B-ultrasound and (or) laparoscopy monitoring during hysteroscopic electro-surgery may help to prevent but not completely avoid uterine perforation. PMID- 12895312 TI - [Clinical analysis of 17 cases of nongestational choriocarcinoma]. AB - OBJECTIVE: To evaluate the diagnosis, treatment and prognosis of nongestational choriocarcinoma. METHODS: Retrospective analysis was done on 17 cases (11 female patients and 6 male patients) of nongestational choriocarcinoma treated in Peking Union Medical College Hospital from 1985 to 2001. RESULTS: All cases received operation as well as chemotherapy. The mean courses of chemotherapy for each patient is 8.6. Correct diagnosis at the first administration was seen in only 3 cases. Nine female and 2 male patients obtained complete remission after the treatment and no recurrence was seen during the 8 to 118 months of follow up. CONCLUSIONS: The diagnosis of nongestational choriocarcinoma is expected to be promoted. Optimal prognosis can be gained for the female patients after treatment by operation as well as chemotherapy, but the prognosis for the male patients is poor. PMID- 12895313 TI - [Epithelial ovarian carcinoma metastatic to the brain: report on ten cases with review of literature]. AB - OBJECTIVE: To investigate the incidence, the diagnosis, the multimodal treatment and the relevant factors of the brain metastases in patients with epithelial ovarian carcinoma (EOC). METHOD: The clinical data were analysed in 10 cases of brain metastases from 478 cases of EOC after treatment between 1996 - 2001. RESULTS: The incidence of brain metastases in EOC was 2.1%. The most common manifestation of the brain metastases was headache, nausea and limb paralysis. The most common metastatic sites were the cupular and occipital part of the cerebra. Seven of ten patients were treated with brain radiation and systemic chemotherapy, three abandoned. The brain irradiation dosage was 30 - 38 Gy for only one focus in the brain lasting for 4 weeks and 40 - 45 Gy for two or three foci in the brain lasting for 5 weeks. The overall survival after identification of brain metastases was < 1 - 33 months, the overall median survival was 6.3 months, but for those who abandoned treatment, the median survival was 1.4 months vs 8.3 months for those who completed the courses of therapy. CONCLUSIONS: The diagnosis of brain metastasis was based on the clinical features and the imagings and the therapy was comprehensive and palliative. However, the number and size of the brain metastases and its reaction to radiation was significantly associated with the prognosis. Although the survival after the identification of brain metastases in EOC is poor, most patients can be palliated successfully by using a combination of radiation therapy and systemic chemotherapy. PMID- 12895314 TI - [Study on the mitochondrion DNA mutation in tumor tissues of gynecologic oncology patients]. AB - OBJECTIVE: To investigate the mitochondrion DNA (mtDNA) mutation in tumor tissues of gynecologic oncology patients and their relationship to tumorigenesis and tumor development. METHODS: The samples of tumor tissue and their proximal normal tissue of 32 gynecological malignant tumor patients as well as 8 gynecological benign tumor patients were taken. The malignants were 5 squamous cervical carcinomas, 10 endometrial carcinomas and 17 epithelial ovarian cancers (EOC). The benign tumors were 4 ovarian epithelial tumors and 4 uterine myomas. Polymerase chain reaction-single strain conformation polymorphism (PCR-SSCP) and DNA sequencing were done to examine mtDNA mutation. RESULTS: The mtDNA mutation rate and polymorphism rate were 68.8% and 56.3% respectively in 32 cases of malignants. The mtDNA mutation rate and polymorphism rate were 2/8 and 4/8 respectively in 8 cases of benigns. The difference between malignants and benigns was significant (P < 0.05). The hot point of mtDNA mutation were located in Cytb gene region. The characteristic mtDNA mutation is multigene and multisites mutation. The mojarity (63.6%) of patients involved in 2 more gene mutation. CONCLUSIONS: There is high frequent mtDNA coding area mutation in gynecological malignant tumors, indicating that mtDNA coding region mutation is closely related to gynecological malignants development and progression. It may be an important extra nuclear molecular genetic alteration in mechanisms of tumorigenesis. PMID- 12895315 TI - [In vivo reversal of multidrug resistance by transduction of human tumor necrosis factor-alpha into drug resistant cell line of choriocarcinoma]. AB - OBJECTIVE: To investigate in vivo reversal of multidrug resistance and biological properties of a drug resistant cell line of choriocarcinoma transduced by human tumor necrosis factor-alpha (TNF-alpha) gene via the establishment of its animal model. METHODS: Choriocarcinoma cell line JEG-3, drug-resistant choriocarcinoma cell line JEG-3/VP2, and human TNF-alpha-transduced drug-resistant choriocarcinoma cell line JEG-3/VP2/TNF-alpha were injected subcutaneously in the neck of nude mices. Tumor size and weight were routinely measured, tumor histological structure was observed and its chemosensitivity was tested. Expression of multidrug resistance (MDR1) mRNA was investigated using reverse transcription-polymerase chain reaction (RT-PCR) and P-glycoprotein (P-gp) expression was determined by immunohistochemistry with the monoclonal antibody MDR1. RESULTS: The rate of inoculation for all three tested cell lines was 100%, the latent period was 10 to 14 days. Tumor growth rates and weights were significantly different among three cell lines (P < 0.05), with the lowest in JEG 3/VP2/TNF-alpha cell line. Tumor inhibition rate after treatment with etopside (VP-16) was significantly higher in JEG-3/VP2/TNF-alpha (41.0% - 42.5%) (P < 0.05), compared with JEG-3/VP2 (24.3% - 28%), and similar to JEG-3 (46.7% 47.7%). Transduction and expression of human TNF-alpha in drug-resistant choriocarcinoma cell line JEG-3/VP2 was found to reverse MDR1 on the mRNA and P gp levels. CONCLUSION: Transduction and expression of human TNF-alpha in drug resistant choriocarcinoma cell line JEG-3/VP2 can reverse expressions of MDR1 mRNA and P-gp, enhance the susceptibility of the JEG-3/VP2 to the cytotoxic drugs, and lower its tumorigenesis. PMID- 12895316 TI - [A multicentre comparative clinical study of uterine cavity shaped device Cu300, TCu220C and TCu380A in ten years]. AB - OBJECTIVE: To observe the clinical effects of uterine cavity shaped device (UCD) Cu300, TCu220C and TCu380A in ten years. METHODS: Two thousand, six hundred and ninety-nine rural parous women were randomized into three groups. UCD Cu300 group had 899 cases, TCu220C group and TCu380A group each had 900 cases. Follow-up visits were performed at 3, 6, 12 months and 2, 3, 5, 10 years after insertion. RESULTS: The cumulative pregnancy rates in the 10th year for UCD Cu300, TCu220C and TCu380A were 2.56, 4.82, and 4.01 per 100 women, respectively. UCD Cu300 group was significantly lower than other two groups (P < 0.05). The expulsion rates were 4.54, 5.46 and 9.68 per 100 women, respectively. TCu380A was significantly higher than UCD Cu300 and TCu220C (P < 0.001). The rates of removal for bleeding and pain for UCDCu300, TCu220C and TCu380A were 6.71, 5.58 and 6.80 per 100 women, respectively, no statistical difference was shown. The cumulative continuation rates were 74.86, 66.22 and 63.56 per 100 women at the 10 th year, UCD Cu300 was the highest, TCu380A was the lowest (P < 0.01). CONCLUSIONS: UCD Cu300, TCu220C and TCu380A are effective for long-term contraception. They are worthy of wide spreading in rural areas. PMID- 12895317 TI - [Analysis of severe acute respiratory syndrome in Beijing]. AB - OBJECTIVE: To study the clinical, laboratory, and radiologic features of 34 cases of severe acute respiratory syndrome (SARS) in Beijing. METHODS: All patients were admitted to the isolation wards. Their demographic, clinical, laboratory, and radiologic characteristics were analyzed. Univariate and multivariate analyses were performed. RESULTS: Eight patients came from a family, and 15 patients were medical staff. The mean age of patients was (33.4 +/- 13.4) years. The latent period varied from 2 to 14 days (median 4 days). The most common symptoms were fever (100%), palpitation (91.7%), myalgia (79.2%), headache (70.8%), diarrhea (73.9%) and cough (58.3%). The mean leucocyte count was (4.6 +/ 1.4) x 10(9)/L, and the mean lymphocyte ratio was 0.27 +/- 0.11. 68.4% of the patients had lymphopenia (absolute lymphocyte count < 1.3 x 10(9)/L). Other common findings included elevated levels of serum alanine aminotransferase, lactate dehydrogenase and erythrocyte sedimentation (76.2%, 28.6% and 47.8%, respectively), and decreased levels of serum iron and albumin (63.2% and 47.8%, respectively). Thirty-two cases had abnormal chest radiographs. In 2 cases in whom typical lung opacities could not be found on the initial plain chest radiographs, thoracic CT proved to be useful. Postmortem examination of 1 patient revealed marked edema with foci of hemorrhage and hyaline membrane formation in the lungs, hemorrhage necrosis and a obvious decline of cells in lymph glands. In a multivariate analysis (Stata 7.0), the independent predictor of an adverse outcome was advanced age (odds ratio per decade of life, 1.6; 95% CI, 1.08 to 2.63; P = 0.007). CONCLUSIONS: Fever, lymphopenia, low serum iron and chest radiograph are helpful to diagnose SARS early; age is the independent predictor of an outcome. PMID- 12895318 TI - [Clinical features and therapy of 106 cases of severe acute respiratory syndrome]. AB - OBJECTIVE: To investigate the clinical characteristics of severe acute respiratory syndrome (SARS) and find out its effective treatment. METHODS: A total of 106 cases of SARS were analyzed prospectively. RESULTS: In this group, 56 were male and 50 female, aged from 15 to 81 years [average (36 +/- 10) years]. Common symptoms included fever (98.1%), chills (75.5%), cough (71.7%), headache and breathless (both 43.4%), diarrhea (24.5%) and rare rales in the lungs (11.2%). Laboratory test showed leukopenia (34.0%), lymphopenia (81.1%) and an extraordinary decrease of CD(4)(+) T cells (98.1%). Other rare abnormalities included liver injury (elevated alanine aminotransferase in 7.6%) and thrombocytopenia (3.8%). Almost all patients suffered from hypoxemia (PaO(2) less than 90 mm Hg in 90.2%, less than 70 mm Hg in 28.6%). Chest radiographs showed that unilateral focal patchy involvement in 34.0% of the patients, and unilateral multifocal or bilateral involvement were 11.3% and 46.2% respectively. Treatment regimens included small doses of steroids (methylprednisolone 40-80 mg, q12 h recommended) accompanied with broad-spectrum antibiotics such as the second generation of cephalosporins and macrolides and some other antiviral drugs. Meanwhile, emphasis was placed on oxygen support and coping with their underlying diseases. CONCLUSIONS: SARS has various presentations of clinical features and laboratory tests. Detection of CD(4)(+) T cell count is beneficial to diagnose SARS in early stage. Effective treatment includes various regimens, oxygen support and small doses of steroids. PMID- 12895319 TI - [Glucocorticoid in the treatment of severe acute respiratory syndrome patients: a preliminary report]. AB - OBJECTIVE: To discuss the effect of glucocorticoid (methylprednisolone) on severe acute respiratory syndrome (SARS). METHODS: Thirty SARS patients were treated at our hospital for over 3 weeks since March to May 2003. The course and dosage of glucocorticoid, counts of CD(4)(+), CD(8)(+) and CD(3)(+), electrolytes, blood routine, and sera albumin before and after the treatment were analysed. RESULTS: Before treatment by methylprednisolone, the counts of CD(4)(+), CD(8)(+) and CD(3)(+) of 27 SARS patients were (401 +/- 203), (340 +/- 187), (756 +/- 383) cells/ microl. Twenty-nine of the 30 SARS patients were treated by methylprednisolone. The dosage for 24 patients was 80 - 160 mg/d with the largest being 1,000 mg/d before admission to the hospital. The count of WBC was increased after treatment (P < 0.01). No obvious effect was observed on the potassium, sodium and chlorine of blood (P > 0.05). Glucocorticoid increased the level of blood glucose (P = 0.01), decreased the level of sera albumin (P < 0.01), and its large dosage decreased the counts of CD(4)(+), CD(8)(+) and CD(3)(+). Three severe patients had secondary infection after administration of a large dose of glucocorticoid. CONCLUSIONS: In the early stage of the disease, the counts of CD(4)(+), CD(8)(+) and CD(3)(+) of SARS patients may reduce markedly indicating the immunity is suppressed. A large dose of glucocorticoid may aggravate the suppression and make the body in an active metabolic state (the increase of blood glucose and the decrease of sera albumin). Thus the disease is aggravated and patients are likely to suffer from severe secondary infection. Indications for use of glucocorticoid must strictly controlled and its large dosage is improper. PMID- 12895320 TI - [The long-term efficacy of lamivudine in chronic hepatitis B: interim analysis of 3-year's clinical course]. AB - OBJECTIVE: To evaluate the long-term efficacy and safety of 3-year lamivudine treatment for chronic hepatitis B and the impact of emergence of YMDD mutation of hepatitis B virus (HBV). METHODS: This multi-center, randomized, double-blind, placebo-controlled trial began in 1996. A total of 429 patients with serum HBsAg, HBeAg and HBV DNA positive were randomized to receive either lamivudine 100 mg daily (n = 322) or placebo (n = 107) in a 3:1 ratio for the first 12 weeks. Thereafter, all patients were offered open label lamivudine 100 mg/d for a total of 156 weeks. RESULTS: After 12 weeks of lamivudine treatment, serum HBV DNA levels decreased rapidly; at week 12 the negativity of HBV DNA (< 1.6 pg/ml) was 92.2%, whereas it was only 14.1% (P < 0.01) in the placebo group. After 1 year of lamivudine treatment, in 72.7% of the patients serum HBV DNA was undetectable (< 1.6 pg/ml). At the end of 3 years, serum HBV DNA continued to be substantially suppressed; the median level was below detectable level in non-YMDD variant patients and was increased to 10 pg/ml in YMDD variant patients. At the end of 1, 2 and 3 years, the HBeAg loss rates were 9.5%, 16.8% and 20.0% respectively; and the HBeAg/anti-HBe sero-conversion rates were 8.3%, 11.5% and 17.3% respectively. The rates of HBeAg loss and seroconversion correlated with baseline ALT levels, in patients with baseline ALT > 2ULN and ALT > 5ULN, the loss of HBeAg was 42.2% and 66.7%, sero-conversion rates were 34.4% and 61.1% respectively (P < 0.01) at the end of year 3. ALT levels at year 3 remained normal in 58.8%, and below baseline in 79.1% of the patients whose ALT were abnormal before treatment. YMDD mutations developed in 12.1%, 49.7% and 70.5% of the patients respectively at year 1, 2 and 3. HBV DNA levels were increased slightly or moderately and accompanied with elevation of ALT. HBeAg loss and sero-conversion could be achieved in YMDD variant patients to 20.0% and 15.1% at the end of year 3, but lower than that in non-variant patients (P < 0.01). The adverse drug reactions or events were generally mild to moderate, 2 patients were reported to have serious events related to the study medication. ALT flares (ALT > 5ULN) occurred in 17 patients, 10 with YMDD variants and 7 with non-variants, but all resolved. No deaths were reported in the 3 year treatment period. CONCLUSION: Sustained HBV replication and clinical improvement could be obtained by 3-year long-term Lamivudine therapy with good tolerance. PMID- 12895321 TI - [A study of the characteristics of pre S/S gene mutation of hepatitis B virus (HBV) in mothers and HBV-infected children via mother-to-infant transmission]. AB - OBJECTIVE: To investigate the characteristics of mutations in pre S/S gene of HBV in asymptomatic carrier (AsC) children infected through mother-to-infant transmission and their AsC mothers with different degree of viremia. METHODS: According to the levels of viremia in every pair of mother and child, 15 pairs of child and mother were divided into three groups 5 pairs in each group in this study: group I (both children and mothers had high viremia), group II (children had high and mothers had low viremia) and group III (children had low and mothers had high viremia). pre S/S gene was amplified by PCR and cloned into pGEM-T vector with T-A cloning technique. The recombinant plasmid pGEM- preS/S was confirmed by digestion with restriction enzyme ApaI and SacI. Tow clones were selected to be sequenced in each patient. The mutations of preS/S were compared with HBV DNA consensus sequence of Chongqing. RESULTS: In each group the subtypes of HBV were B/adw2 in 4 pairs and C/adrq+ in one pair. The preS/S clones in patients infected with subtype B/adw2 HBV were analyzed. It was shown that there was no difference among the four high viremic groups or between the two low viremic groups in the number of mutation and the mutational position. However, there was significant difference between the high viremic group and low viremic group. The mutation was not related to age. In the two low viremic groups (the mothers of group II and the children of group III), there were 86-94 mutational positions in 13/16 clones. There were 86 same mutational positions causing 37 amino acid changes in 11/13 clones and 90 same mutational positions causing 38 amino acid changes in 2/13 clones. Most of the changed amino acids were located within T and B cell epitopes of the envelope protein or/and the surrounding regions. Sixty-two mutational positions that resulted in 28 amino acid changes were same in these two mutational sequences. CONCLUSIONS: The mutation of HBV is not associated with the duration of infection. There are many differences of mutation when HBV comes from a same strain in hosts with different degrees of viremia. There are some regular patterns in the mutation of HBV after occurrence of HBeAg seroconversion. The mutation could be related to the escape of the attack of host's immunity. PMID- 12895322 TI - [Hepatitis C virus nonstructural 5A protein inhibits tumor necrosis factor alpha mediated apoptosis of HepG2 cells]. AB - OBJECTIVE: To study the suppressive effect of HCV nonstructural 5A (NS5A) protein on tumor necrosis factor alpha (TNF alpha) mediated apoptosis of HepG2 cells. METHODS: NS5A gene fragment was amplified by reverse transcription and nested polymerase chain reaction from serum samples positive for anti-HCV. The PCR product was directly cloned using TA cloning kit, and 2 independent clones were isolated, digested and sequenced. Then we constructed HCV NS5A expression plasmid (pcDNA3.1-NS5A), stably transfected into HepG2 cells with lipofectamine. Successful transfection of NS5A gene and expression of NS5A protein were confirmed by Western blot. Transfected cells were incubated with TNFalpha for 48 h, then labeled with Annexin V and visualized by fluorescence microscopy. To examine the effects of NS5A protein on the apoptotic signaling pathway, caspase-3 cleavages and release of cytochrome C were investigated in the transfectant treated with TNF alpha for 48 h and the cell cytosol was subjected to SDS-PAGE and Western blot analysis. RESULTS: The stable transfectant of HepG2 cells lines for HCV NS5A protein expression was achieved. The NS5A protein blocked the activation of caspase-3 and the release of cytochrome C in the TNF alpha treated cells. CONCLUSION: HCV NS5A protein inhibits TNF alpha mediated apoptosis of HepG2 cells in vitro. PMID- 12895323 TI - [Changes in peripheral lymphocyte subfraction in cytomegalovirus infections during the early stage of allogenic peripheral blood stem cell transplantation]. AB - OBJECTIVE: To study the alterations and the clinical significance of peripheral lymphocyte subsets in patients with active cytomegalovirus (CMV) infection during the early stage of allogeneic peripheral blood stem cell transplantation (PBSCT) and to investigate the influence of active CMV infection on immunity. METHODS: K(3)-EDTA anticoagulated peripheral blood samples were collected weekly from 27 allogeneic PBSCT recipients in the first 3 months after PBSCT. CMV pp65 antigen in peripheral white blood cells were tested by indirect immunofluorescence assay, and lymphocyte subsets were detected by flow cytometry with specific fluorescent monoclonal antibodies. In the meantime lymphocyte subsets of 51 samples from healthy blood donors were tested as normal control values. RESULTS: CD(4)(+) T cell count for all recipients was significantly low throughout the first 3 months after PBSCT as compared with normal donors (P < 0.01). Of the 27 allogeneic PBSCT recipients, 5 had no active CMV infection, 10 had asymptomatic CMV active infection, 12 had symptomatic CMV infection. The average CD(4)(+) T cell counts (x 10(6)/L) for the different groups of patients were 328 +/- 203, 239 +/- 218 and 199 +/- 92; The average CD(8)(+) T cell counts (x 10(6)/L) were 400 +/- 380, 267 +/- 206 and 603 +/- 461, respectively. The average percentage of CD(4)(+) CD(28)(+) T cell subsets were (89.2 +/- 8.9)%, (84.2 +/- 10.1)% and (63.5 +/- 11.4)%, respectively. The recipients with asymptomatic CMV active infection were likely to have more natural killer (NK) cells as compared with those without active CMV infection (P < 0.01). The patients with symptomatic CMV infection had significantly lower CD(4)(+) cell count and higher CD(8)(+) cell count than those with asymptomatic CMV infection (P < 0.01). The percentage of CD(4)(+) CD(28)(+) cells was decreased significantly in the CMV active infection group (P < 0.01). CONCLUSIONS: CMV active infection could induce dramatic alterations in lymphocyte subsets lymphocyte subsets and host immunity for allogeneic PBSCT recipients. Therefore, the changes of lymphocyte subsets might serve as auxiliary parameters to predict active CMV infections in this kind of immunocompromised patients. PMID- 12895324 TI - [Effect of radiofrequency ablation on endothelial function and platelet activation]. AB - OBJECTIVE: To investigate the effect of radiofrequency catheter ablation on endothelial function and platelet activation. METHODS: With radioimmunoassay, enzyme linked immunosorbent assay, cell labeling with monoclonal antibody and flow cytometry technique, thirty-one consecutive patients were checked for levels of plasma endothelin (ET), von willebrand's factor (vWF) and expression of platelet alpha-granule membrane glucoprotein (CD(62P)), platelet lysosome membrane glucoprotein (CD(63)) before, immediately and 47 - 115 hours after radiofrequency catheter ablation. RESULTS: There was no significant difference in the levels of plasma ET and vWF before and after catheter ablation; The rate of CD(62P) and CD(63) expression rate on platelets was (4.75 +/- 2.32)% and (9.62 +/ 4.08)% before catheter ablation, and it significantly increased to (7.64 +/- 5.25)% (t = 3.05, P < 0.01) and (12.23 +/- 5.70)% (t = 2.10, P < 0.05) immediately after catheter ablation and then returned to baseline levels 47 - 115 hours after catheter ablation. There was a positive correlation between the change of CD(62P) expression and total energy dose of radiofrequency (r = 0.30, P < 0.05). CONCLUSIONS: Radiofrequency catheter ablation did not bring about significant damage to endothelium but can increase expression of platelet membrane CD(62P), CD(63) and promote platelet activation, total energy dose of radiofrequency is one of important influencing factor. PMID- 12895325 TI - [Losartan downregulates the expression of transforming growth factor beta type I and type II receptors in kidney of diabetic rat]. AB - OBJECTIVE: To investigate the influence of losartan on the expression of transforming growth factor-beta type I and type II receptors (TGF beta RI, TGF beta RII) in kidney of diabetic rat model. METHODS: The Wistar rats were randomly divided into 3 groups with 10 rats in each group: normal control group (group C), diabetic rat model group (group DM, the model was reproduced by intravenous injection of 50 mg/kg streptozotocin), and the group of diabetic rat model treated with losartan (group DL, 10 mg.kg(-1).d(-1) of losartan intragastrically since the 2nd day of streptozotocin injection). Eight weeks later, the mRNA and protein expressions of TGF beta RI, TGF beta RII and fibronectin in renal cortices of all 3 groups were measured by semiquantitative reverse transcription polymerase chain reaction and by histoimmunochemical methods. The blood glucose, urea and creatinine were detected by biochemical assay and the blood insulin and angiotensin II by radioimmunoassay. The urine albumin excretion ratio was examined by sulfosalicylic acid assay. RESULTS: The mean glomerular volume, kidney weight/body weight ratio, urine albumin excretion ratio, blood urea and creatinine were significantly elevated in group DM than that in group C (P < 0.05). The mRNA and protein expressions of TGF beta RI, TGF beta RII and fibronectin in renal cortices of group DM were significantly increased than that in group C as well (P < 0.05). The increase in mean glomerular volume, kidney weight/body weight ratio, urine albumin excretion ratio, blood urea and creatinine in group DL was significantly attenuated as compared to group DM. The mRNA and protein expressions of TGF beta RI, TGF beta RII and fibronectin in renal cortices of group DL were also significantly reduced as compared to that in group DM (P < 0.05). CONCLUSION: Losartan may have protective effects on the kidney of diabetic rat model. The mechanism of the protection might related to its downregulatory effect on the expression of TGF beta system, which inhibited the hypertrophy of kidney cells and reduced the synthesis of extracellular matrix. PMID- 12895326 TI - [The early diagnosis and treatment of gastric mucosa-associated lymphoma]. AB - OBJECTIVE: To determine the general markers for the diagnosis of gastric mucosa associated lymphoid tissue (MALT) lymphoma and select optimal cases to be interfered in early stage. METHODS: To investigate the changes of histology, Ki 67 expression, monoclonality of IgH gene rearrangement in gastric lymphoid hyperplasia (GLH) cases before and after Helicobacter pylori (Hp) eradication treatment. Histological changes were classified according to Isaacson's criteria of GLH; L26, UCHL-1, anti-Kappa, anti-Lambda and Ki-67 were applied for immunohistochemical staining; semi-nest PCR was used to detect IgH gene rearrangement. RESULTS: Thirty-one cases of GLH with predominant chronic gastritis and gastric ulcer were studied. The ratio of male to female in the 31 cases was 1.8 and the mean course 6.8 years. Twenty-nine cases had Hp infection. Most of the cases belonged to histological grade II and III. Only 1 case had lambda light chain restriction and 10 cases had monoclonal IgH gene rearrangement. Expression of Ki-67 and monoclonal IgH gene rearrangement were significantly increased with the crescendo of lymphoid hyperplasia grading (P < 0.05). 28 cases were given antibiotics with eradication of Hp in 24 cases and 2 cases had to be given another course of treatment. These cases were followed-up on average for 4.6 months. 18 cases showed complete regression both histologically and endoscopically, 4 cases partial regression and 6 cases no change. Four cases with histological grade II, III had the decrease of expression of Ki-67 and reversed to polyclonal IgH gene rearrangement, while there was no change in 4 cases with histological grade IV. CONCLUSIONS: General considerations of histology, expression of Ki-67 and combined clonality of IgH gene rearrangement help to screen cases which need intervention early and make a diagnosis of gastric MALT lymphoma in early stage. Monoclonal IgH gene rearrangement in cases with histological grade III or IV appears to carry more clinical significance. PMID- 12895327 TI - [The role of intercellular adhesion molecule-1 in binding of acute myeloid leukemic blasts cells to human umbilical vein endothelial cells]. AB - OBJECTIVE: To evaluate the adhesion of acute myeloid leukemia (AML) blasts to human umbilical vein endothelial cells (HUVECs) and the roles of intercellular adhesion molecule-1 (ICAM-1) and its ligand lymphocyte function associated antigen-1 (LFA-1) in binding of leukemic blasts to HUVECs. METHODS: AML blasts attachment to unactivated or tumor necrosis factor alpha (TNF alpha) activated endothelial cell monolayers was investigate in vitro; The adhesion of leukemic blasts co cultured with unactivated endothelial cells under static conditions at 37 degrees C for 24 hours was observed;The binding of neutrophils and unactivated endothelial cell monolayers exposed to supernatant of blasts were tested. ICAM-1 on endothelial cell surface and sICAM-1 of endothelial cell supernatant were determined by flow cytometry and ELISA detection. We also observed the adhesion of leukemic blasts in the presence of the adhesion blocking mAbs anti-ICAM-1 and anti-LFA-1. RESULTS: This study has shown that the blast cells attached to unactivated endothelium was little (24.33 +/- 2.87)% and increased after exposure of endothelium to TNF alpha (81.87 +/- 4.08)% (n = 21, P < 0.001); The binding of blasts to endothelium also increased significantly after 24 hours co cultured with unactivated HUVEC (82.06 +/- 7.05)%, (n = 21, P < 0.001); The adhesion of neutrophils and unactivated endothelial cell monolayers exposed to supernatant of blasts was increased significantly (83.99 +/- 3.86)%, (n = 21, P < 0.001). Lower levels of ICAM-1 and sICAM-1 expression was detected on unactivated HUVECs (55.81 +/- 4.11)%, (0.839 +/- 0.236) microg/L respectively. Treatment of HUVECs with AML blasts supernatant for 24 hours increased the expression of ICAM-1 (65.36 +/- 5.97)%, (1.424 +/- 0.469) microg/L respectively (n = 21, P < 0.05) and anti-ICAM 1 and anti-LFA-1 significantly inhibited the adhesion of AML blasts attachment to TNF alpha activated-endothelial cell monolayers (20.12 +/- 1.73)%, (n = 10, P < 0.001). CONCLUSION: Our results indicate that leukemic blasts have the ability to generate some factors that stimulate endothelial cell to secrete ICAM-1 ane to promote their own adhesion to vascular endothelium, interaction of ICAM-1 and its ligand LFA-1 has a key role in adhesion of leukemic blasts and HUVECs. PMID- 12895328 TI - [The therapeutic effects of FLAG regime for refractory and relapsing acute leukemia]. AB - OBJECTIVE: To investigate the therapeutic effects of FLAG regimen for refractory and relapsing acute leukemia. METHODS: Fludarabine 30 mg.m(-2).d(-1), 3 - 5 days, Ara-C 1.5 g.m(-2).d(-1), 5 d and, G-CSF 5 microg.kg(-1).d(-1) were used to treat 7 cases (10 times) of refractory and relapsed leukemia. RESULTS: Of the 10 times, nine showed response to therapy; one case died of complication. Neutropenia begin at the time of 5 - 12 days after the regimen initiating and last 5 - 34 days, thrombocytopenia begin at 5 - 18 days and last 5 - 36 days. The adverse reaction includes hyperamino transferase, pyrexia and diarrhea. CONCLUSION: FLAG shows obvious effects for refractory and relapsing acute leukemia and gains time for hematopoietic stem cell transplantation. PMID- 12895329 TI - [Effects of mifepristone of different doses on emergency contraception, a randomized double-blind study]. AB - OBJECTIVE: To compare the effects of mifepristone of different doses on emergency contraception. METHODS: 3,052 healthy women with regular menstrual cycle who visited the 10 family planning institutes and hospitals in Beijing, Shanghai, Shangdong, Sichuan, Tianjin, Guangdong, and Liaoning for emergency contraception within the period of 120 hours after a single act of unprotected sex were given a single dose of 10 mg or 25 mg mifepristone randomly and double-blindly. They were asked to record the vaginal hemorrhage that would occur and not to have unprotected sex until the next menstrual onset when they were followed up. The trial for a specific subject ended when she menstruated. If the menstruation was irregular or a specific subject failed to menstruate on time a blood or urine human chorionic gonadotropin (hCG) test was made. If the hCG test was negative, an appointment was made to follow up once one week later. If the hCG test was positive ultrasound examination was made to detect pregnancy. If the subject still failed to menstruate and the hCG test was still negative follow-up for this subject could be finished. RESULTS: Twenty-two of the 3,052 subjects were lost to follow up. Among the remaining 3,030 women 1,516 were in the 10 mg group and 1 514 in the 25 mg group. Seventeen pregnancies occurred in each group, with a pregnancy rate of 1.1% for both groups. The relative risk of pregnancy of treatment of 25 mg mifepristone in comparison with treatment of 10 mg mifepristone was 1.0 (95% CI: 0.51-1.95). Both doses prevented about 85% approximately 86% of the anticipated pregnancy if no measure had been adopted. The pregnancy rate nearly doubled in the women who had unprotected sex after treatment of mifepristone. The efficacy of mifepristone decreased along with the delay of mifepristone administration. Side effects were uncommon and mild. Delay of 7 days or more in the onset of next menstruation occurred in 9%-10% of the women. CONCLUSION: Mifepristone of the dose of 10 mg is safe and effective for emergency contraception. Earlier administration is preferable, although the method can be used effectively up to five days after the unprotected sex. PMID- 12895330 TI - [The study on endometrial estrogen receptor and progesterone receptor isoforms in medical abortion women]. AB - OBJECTIVE: To determine the expression and location of estrogen and progesterone receptor isoforms in the endometrium after abortion and discuss the relationship between the changes of isoforms and prolonged uterine bleeding after medical abortion. METHODS: 45 endometrium biopsy were obtained from 15 women with prolonged bleeding after medical abortion, 15 women with prolonged bleeding after vacuum aspiration and 15 without prolonged bleeding after vacuum aspiration as control group. Immunohistochemical staining intensity of ER alpha, ER beta, PR-A and PR-B in endometrium were compared among these groups, as medical abortion group, surgical abortion group and control group. RESULTS: The endometrial ER alpha, ER beta, PR-A and PR-B H-Scores of medical abortion group were 166.7 (88.5), 161.5 (84), 180.0 (109) and 136.5 (119.0), The endometrial ER alpha, ER beta, PR-A and PR-B H-Scores of control group were 306.7 (45.9), 230.0 (75.0), 224.2 (57) and 389.6 (106.6). The endometrial ER alpha, ER beta, PR-A and PR-B H Scores of surgical abortion group were 109.4 (114.6), 225.0 (120.0), 175.5 (225) and 50.0 (89.0). The endometrial ER alpha, ER beta, PR-B H-Scores of medical abortion group were significantly lower than those of control group. And in surgical abortion group ER alpha and PR-B H-Scores were significantly lower than those in control group. But there was no significant difference between ER beta H Scores of surgical abortion group and control group. There were no significant differences of ER and PR isoforms level ratios of endometrial glands to endometrial stroma among three groups. CONCLUSIONS: The decreased ER alpha, ER beta and PR-B levels in the human endometrium may be related to prolonged uterine bleeding after medical abortion by mifepristone accompanied with misopristol. The endometrial ER alpha, PR-B decreased in women after vacuum aspiration abortion may be related to abnormal prolonged uterine bleeding. The changes of endometrial ER, PR isoforms in glands and stroma were simultaneous after medical abortion and surgical abortion. PMID- 12895331 TI - [Effects of long-term installation of intrauterine devices on intrauterine microenvironment]. AB - OBJECTIVES: To study the effect of long-term installation of intrauterine devices (IUD) on the intrauterine microenvironment. METHODS: Eighty-nine healthy 26 - 50 year-old women undergoing physical examination or having their IUD removed were recruited. Among them 62 had used IUDs, including 32 inert IUD (I-IUD) and 30 copper releasing IUD (T-IUD), for 5 - 20 years, and 27 women without installation of IUD were used as controls. In the 3rd to 13th day of menstrual cycle, 3 ml of irrigation of intrauterine cavity were collected to examine the concentrations of tumor necrosis factor-alpha (TNF-alpha), interleukin 2 (IL-2) and secretary IgA (SIgA) by radioimmunoassay (RIA). Hysteroscopy was used to obtain 3 pieces of endometrium at the place where the IUD was attached for each subject. Three pieces of endometrium were curretted from each control during the proliferative stage. Thirty specimens of endometrium, including 10 cases with I-IUD, 10 cases with T-IUD, and 10 control cases were used to examine the distribution of T cell subset by immunohistochemistry. Fifteen specimens of endometrium, including 6 cases with I-IUD, 6 cases with T-IUD, and 3 control cases, were used to examine the endometrial ultrastructure by scanning electron microscopy (SEM) and transmission electron microscopy (TEM). Twenty-five specimens of endometrium, including 10 cases with I-IUD, 10 cases with T-IUD, and 5 control cases, were used to detect the expression and mutation of P16, P53 and K-ras by PCR-SSCP technique. RESULTS: There was no significant difference in TNF-alpha level in the irrigation among the three groups (P > 0.05). IL-2 and SIgA levels (0.39 +/- 0.18) microg/L and (2000 +/- 1224) microg/L respectively) in the T-IUD group were significantly lower than those in the control group (0.96 +/- 0.15) microg/L and (3377 +/- 1906) microg/L respectively, both (P < 0.05). There was a significant difference in the population of T lymphocytes (CD(8)(+) and CD(4)(+) lymphocytes) between the T-IUD group and control group (P < 0.05). SEM and TEM showed no necrosis and atypia in the endometrial cells of I-IUD group and T-IUD group. There was no positive expression of gene p16, p53 and K-ras in the endometrium of the three groups. CONCLUSION: It is safe and effective using I-IUD or T-IUD for a long time, however, T-IUD has some effects on intrauterine local immune function. PMID- 12895332 TI - [Apparent protective effect of high density lipoprotein against coronary heart disease in the elderly]. AB - OBJECTIVE: To evaluate the relationship between the level of high-density lipoprotein cholesterol (HDL-C) and acute myocardial infarction (AMI) events and coronary heart disease (CHD) death, and to explore the protective effect of high level HDL against CHD in the elderly Chinese. METHOD: A prospective study was done upon 1211 retired cadres, 92% of which were males, enrolled in succession since 1986, aged 70 +/- 9 on average when being enrolled and aged 80 +/- 9 on average by the end of the study. During a follow up study with an average duration of 11.2 years, till the year 2000 or a specific subject died of CHD or other disease, all the participants received yearly physical examination and blood chemistry survey. CHD risk factors were screened by logistic regression analysis. According to their HDL-C levels, the subjects were divided into 3 groups: low HDL-C (< 1.03 mmol/L), medium HDL-C (1.03 - 1.56 mmol/L), and high HDL-C (> 1.56 mmol/L) groups. The differences in AMI event and CHD death in each group were analyzed. RESULTS: 411 of the subjects were with normal blood lipid (normal blood lipid group, total cholesterol < 5.17 mmol/L, triglyceride < 1.69 mmol/L), 338 subjects were basically healthy (basically healthy group), and 214 subjects developed CHD during the follow-up (CHD group). 23.2% of the subjects could be included in the low HDL-C group, 68.7% in the medium HDL-C group, and 68.5% in the high HDL-C group. The low HDL-C rate was 16.9% in the basically healthy group and 35.0% in the CHD group (P = 0.000165). The high HDL-C rate was 12.4% in the basically healthy group and 3.3% in the CHD group (P = 0.000645). During the follow up study, 214 cumulative attacks of acute coronary syndrome (mostly AMI) were recorded with 89 deaths. 308 cases died of other diseases. The AMI event rate and CHD death rate in the normal HDL-C group were lower than those in the low HDL-C group by 40% and 53% respectively. The AMI event rate and CHD death rate in the high HDL-C group were lower than those in the normal HDL-C group by 56% and 50% respectively. In the normal lipid group, AMI event rate and CHD mortality in the individuals with low HDL-C level were lower than those in the individuals with medium HDL-C level by 79% and 77% respectively (both P < 0.0001). The number of individuals with high HDL-C level was low, however, no CHD death occurred among these subjects. CONCLUSION: Low HDL is an important independent risk factor of AMI attacks and CHD death in the elderly. High HDL protects effectively against coronary artery disease. High TC level weakens the protective effect of HDL-C. PMID- 12895333 TI - [Operative treatment of basilar-vertebral aneurysms, analysis of 153 cases]. AB - OBJECTIVE: To evaluate the effect of surgery and conservative treatment on basilar-vertebral aneurysms. METHODS: The clinical data of 153 cases with basilar vertebral aneurysms treated in Tiantan Hospital from Jan. 1990 to Oct. 2001 were analyzed retrospectively. Thirty-eight untreated patients with basilar-vertebral aneurysms and one patient with cerebral posterior artery aneurysms were followed up by telephone and letters for 0.42 to 7.92 years, with the mean of 1.91 years. RESULTS: Of the 1 232 patients with intracranial aneurysms, 153 (12.4%) cases, 88 males and 65 females, aged 2 - 69 with a mean age of 40.75 years, suffered from basilar-vertebral aneurysms. The peak of incidence was 40-50 years. One hundred and two cases (66.7%) received surgical treatment. Excellent outcome was observed in 60 cases (58.8%), good outcome in 10 (9.8%); 29 cases (28.4%) showed a bad outcome, and 3 patients died with a mortality rate of 2.9%. Fifty-one cases received conservative treatment. After the surgery, 13 cases died of rupture of aneurysms during hospitalization. Eight (61.5%) aneurysms were located in basilar artery, 2 (15.4%) in cerebral posterior artery, 1 (7.7%) in vertebral artery, 1 in basilar-vertebral junction, and 1 in cerebella inferior posterior artery. According to Hunt and Hess classification, 2 cases were considered of degree 0, 1 case degree 1, 5 cases degree 2, 1 case degree 3, and 4 cases degree 4. The age range was 5 - 69 years and the mean age was 38.69 years of the above 13 patients, 10 cases (76.9%) had previously suffered from subarachnoid hemorrhage, 2 suffered from the attack twice. 3 cases (23.1%) with unruptured aneurysms had symptoms. Among the 38 followed-up patients without surgical treatment, 3 patients (17.6%) with the mean age of 52.67 years, of which 2 had aneurysms located in basilar artery and one in cerebral posterior artery, died of aneurysm rupture 0.42, 0.72 and 1.5 years after discharging respectively. Before death two cases presented symptoms and one died accidentally. CONCLUSION: Active surgical treatment effectively decreases the high mortality rate of basilar-vertebral aneurysms. PMID- 12895334 TI - [Application of ultrasonic angioplasty in treating totally occluded peripheral arteries, a clinical study of 39 cases]. AB - OBJECTIVE: To evaluate the efficacy and safety of intravascular ultrasonic angioplasty in treating atherosclerotic totally occluded peripheral arteries. METHODS: Percutaneous ultrasonic angioplasty was applied on 39 cases with peripheral atherosclerotic total occlusion under fluoroscopic guidance. The recanalization, clinical efficacy and complications were observed. RESULTS: 38 (97.4%) of the 39 cases achieved successful recanalization in (1157 +/- 376) seconds, with a mean residual stenosis of (32 +/- 11)%. The technical successful rate (residual stenosis < 50%) was 82.1% (32/39). Clinical improvement at different degrees was seen in 38 cases after ultrasonic angioplasty. The patency rates 24 hours, 30 days, 90 days and 180 days after the ultrasonic angioplasty were 100%, 94.7%, 89.5% and 81.6% respectively. The complications related to ultrasonic angioplasty included hemoglobinuria in 9 cases and distal embolization in 1 case. CONCLUSION: Ultrasonic angioplasty appears to be safe and effective for the treatment of atherosclerotic totally occluded peripheral arteries. PMID- 12895335 TI - [Relative stasis of hepatitis C virus quasispecies in natural duration]. AB - OBJECTIVE: To investigate the evolution of HCV quasispecies in persistent and self-limited infection patients. METHODS: Peripheral blood HCV preserved for 10 years from 8 naive patients were analyzed. HCV core and E2 genes were cloned and sequenced. Genetic distances of different isolates at different time were analyzed by phylip software. RESULTS: The genetic distances of HCV quasispecies were higher in persistent infection patients than that of self-limited patients. In each patient the genetic distance of HCV E2 region isolates were higher than that of HCV C region which has some relationship between each other. In the 4 persistent patients there exist no differences between within-group and between group genetic distance in HCV C region and also exist no differences in HCV E2 region in 2 of the 4 persistent patients. CONCLUSIONS: The diversity degree of HCV quasispecies may affect the outcome of hepatitis C. Research on HCV C and E2 region to study HCV quasispecies may obtain different results. The distribution of HCV quasispecies may be stable in hepatitis C natural history. PMID- 12895336 TI - [Relationship among the XhaI and EcoRI locus polymorphisms of apolipoprotein B gene, serum lipid metabolism and gallstone disease]. AB - OBJECTIVES: To explore the relationship between the XbaI and EcoRI locus polymorphisms of apolipoprotein B gene and gallstone disease. METHODS: Restriction fragment length polymorphism-polymerase chain reaction (RFLP-PCR) technique was used to analyze the genotype of the ApoB gene in 106 patients and 105 controls, according to the design of case control study. RESULTS: The frequencies of X+X- and X-X- of XbaI locus polymorphism were significantly different between the patients and controls and the frequency of X+ allele in the patients was significantly higher than that in the controls (0.104 vs 0.052). Meanwhile, the levels of LDLc and ApoB in the patients were significantly higher than those in the controls among the group of X+X- genotype. The frequencies of E+E- and E+E+ of EcoRI locus polymorphism were significantly different between the patients and controls and the frequency of E-allele in the patients was significantly higher than that the in controls, and the level of LDLc with E+E- genotype was higher than that with E+E+ genotype among the patients. CONCLUSION: ApoB gene X+ allele of XbaI locus and E-allele of EcoRI locus may be the susceptible genes for gallstone disease, and variation of X+ and E-alleles may affect serum lipid metabolism and formation of gallstone. PMID- 12895337 TI - [Construction of a non-viral vector H1s-EGFc and a preliminary study on its function]. AB - OBJECTIVE: To construct a non-viral vector for targeting cancer gene therapy. METHODS: The coding sequence of H1s-EGFc was inserted into the expression vectors of Pichia pastoris, and the fusion protein was expressed in secretary way. H1s EGFc was purified by anion exchange chromatography and size exclusion chromatography. H1s-EGFc fusion protein and "killing gene" expression recombinant pKG plasmid DNA were dissolved in serum-free RPMI-1640 culture to produce H1s EGFc/pKG complex. HeLa cells, an epidermal growth factor receptor (EGFR) highly expressing cell line, and Jurkat cells, an EGFR non-expressing cell line, were cultured and transfected with H1s-EGFc/pKG complex of different concentrations. Trypan blue staining was used to calculate the number of live cells and the killing rate of H1s-EGFc/pKG. RESULTS: H1s-EGFc fusion protein was constructed and expressed with a purity of over 90%. When the concentrations of H1s-EGFc/pKG complex were 3 microg/ml, 6 microg/ml, and 9 microg/ml respectively the killing rates were 30.6%, 36.2%, and 58.1% respectively. CONCLUSION: The fusion protein H1s-EGFc binds functional gene efficiently and targets it into specific cells. It can be used as non-viral vector in target cancer gene therapy. PMID- 12895338 TI - [Cloning and expression of insulin-like growth factor-1 platelet-derived growth factor-AA, and tumor growth factor beta1 genes in adenovirus vector]. AB - OBJECTIVE: To establish recombinant adenovirus highly expressing genes of insulin like growth factor-1 (IGF-1), platelet-derived growth factor-AA (PDGF-AA), and tumor growth factor beta1 (TGF beta 1) to be used in gene therapy. METHODS: Normal human osteoblasts were obtained from cancellous bone of lilium left from bone grafting and then cultured. Total RAN was extracted and IGF-1, PDGF-AA, and TGF beta 1 cDNAs was obtained by RT-PCR. Vector p-Shuttle containing these cNDAs was cloned into adenovirus and then the recombinant adenovirus was transfected into HEK293 cells. Recombinant adenovirus containing reporter gene LacZ was used in control group. Western blotting was used to detect the expression of these growth factors. Human osteoblasts were cultured and transfected with recombinant adenovirus. MTT method was used to detect the proliferation of the cells. Paranitrophenol method was used to examine the activity of alkaline phosphatase (ALP) in osteoblasts. RESULTS: Expression of IGF-1, PDGF-AA, and TGF beta 1 cDNA and expression of IGF-1, PDGF-AA, and TGF beta 1 proteins were found in HEK293 cells transfected with the recombinant adenovirus and not in the control HEK293 cells. The proliferation and ALP activity of osteoblasts transfected with the recombinant virus were significantly increased in comparison with those of the control osteoblasts (all P < 0.010). Immunohistochemical staining showed significant brown particles in the osteoblasts transfected with the recombinant virus and none in the control osteoblasts. CONCLUSION: The recombinant adenovirus thus constructed expresses the proteins of several growth factors with bioactivity in human cells and can be used as a satisfactory gene tool. PMID- 12895339 TI - [Effect of arsenic trioxide on rabbit vascular smooth muscle cells after balloon angioplasty]. AB - OBJECTIVE: To study the apoptotic effect of arsenic trioxide on rabbit vascular smooth muscle cells (RVSMCs) after balloon angioplasty. METHODS: A New Zealand rabbit underwent balloon angioplasty three times at one of its iliac arteries. The iliac artery at the opposite side was used as control. Three days after the rabbit was killed. The VSMCs of the iliac artery undergoing balloon angioplasty were cultured. Arsenic trioxide of different concentrations was added into the culture. VSMCs not co-incubated with arsenic trioxide was used as controls. Cell proliferation curve was drawn. DNA was extracted and underwent electrophoresis. Transmission electron microscopy was used to observe the ultramicroscopic structure. TUNEL technique was used to examine the apoptosis of cells. RESULTS: Cell proliferation curve showed that the control cells grew geometrically and reached a plateau stage by day 3. The cells co-incubated with arsenic trioxide proliferated more slowly and the plateau stage appeared earlier dose-dependently (P < 0.05). Electrophoresis showed obscure ladder band of DNA specific to cell apoptosis, especially at the concentrations of arsenic trioxide of 3.0 and 4.0 micro mol/L. Transmission electron microscopy showed apoptotic cells with apoptosis nodes. TUNEL technique showed that the number of yellow-dyed cells was significantly larger after the addition of arsenic trioxide. The above-mentioned manifestations were not seen in the control group. CONCLUSION: Arsenic trioxide increases the apoptosis of rabbit vascular smooth muscle cells after balloon angioplasty and inhibits RVSMCs over-proliferation. It is a hopeful measure in prevention of re-stenosis after angioplasty. PMID- 12895340 TI - [Construction, expression and antigenic study of bivalent vaccine candidate with 26,000 OMP and heat short protein A of human Helicobacter pylori]. AB - OBJECTIVE: To construct a recombinant vector containing gene encoding heat shock protein A (HspA) and outer membrane protein (OMP) with relative molecule mass (Mr) of 13,000 and 26,000 respectively from human Helicobacter pylori (Hp) and be expressed in E. coli BL21, as well as analyse its antigenic for the exploiting vaccine of Hp. METHODS: The target gene encoding heat shock protein A was amplified from Hp chromosome by PCR. And then digested by restricted endonuclease enzyme of kpnI, BamH I simultaneously, and inserted into the prokaryotic expression vector pET32a(+) digested by corresponding restricted endonuclease enzyme. The recombinant vector was used to select and transform for sequence analysis. After pET32a(+)/HspA and pET32a(+)/Omp(26) digested by restricted endonuclease enzyme of Hind III, BamH I simultaneously, the pET32a(+)/HspA and 26,000 OMP were taken out of agarose electrophoresis, and connected by T4 ligase. The recombinant vector pET32a(+)/HspA-Omp(26) was used to select and transform, meanwhile expressed in E. coli BL21(DE3). The antigenic of recombinant fusion protein was analysed by western blotting. RESULTS: Enzyme digestion analysis and sequencing showed that the target genes was found to be 951 base pairs, and had been inserted into recombinant vector, but as compared with gene reported by GenBank, 1.15% of the gene mutation and 1.26% of amino acid residues change in Hp happened respectively. SDS-PAGE analysis showed that recombinant vector could be expressed in E. coli BL21, its relative molecule mass of expressed product was 59 x 10(3), while Mr of protein expressed by pET32a(+) of them was about 20 x 10(3), and soluble expression product accounted for 19.96% of total bacterial protein. After purification with Ni-NTA agarose resin, the purity of recombinant fusion protein was about 95%. The western blot result showed that recombinant fusion protein could be recognized by anti-Hp positive serum and monoclonal antibody of 26,000 OMP, suggesting that this protein had good antigenic. CONCLUSION: The genes coding HspA and OMP with Mr 13,000 and 26,000 respectively are cloned and expressed successfully. The results obtained lay the foundation for research on development of Hp protein and DNA vaccine and a quickly diagnostic kit applying to detection of Hp infection. PMID- 12895341 TI - [The relationship between tumor suppressor genes p14ARF and p53 expression and biological behavior of pancreatic carcinoma]. AB - OBJECTIVE: To investigate the relationship between tumor suppressor genes p14(ARF) and p53 expression and the biological behavior of pancreatic carcinoma. METHODS: Modified Envision method of immunohistochemistry was used on 42 specimens of pancreatic adenocarcinoma and 10 normal specimens of normal pancreas resected during operation to examine the expression of the gene p14(ARF). and ABC immunohistochemical technique was used to examine the expression of p53. RESULTS: The positive rates of p14(ARF) in of normal pancreatic tissues and pancreatic carcinoma were 90% and 35.7% respectively (P < 0.01). The positive rates of p53 in tissues of normal pancreatic and pancreatic carcinoma were 0 and 42.5% (P < 0.05) respectively. Significant correlation was detected between the expression of p14(AR) and p53 and the diameter of tumor, the rate of lymph node metastasis, pathological grade, and clinic stage in pancreatic carcinoma. No correlation was found between the expression of p14(ARF) and p53 and the tumor infiltration in pancreatic carcinoma. CONCLUSION: The tumor suppressor gene p14(ARF) and p53 are closely related to the occurrence and development of pancreatic carcinoma. It is possible to treat pancreatic carcinoma by gene intervention. PMID- 12895342 TI - [Transcranial Doppler monitoring of cerebral blood flow velocity during extracorporeal membrane oxygenation in sheep]. AB - OBJECTIVE: To investigate the hemodynamics during extracorporeal membrane oxygenation (ECMO) and the value of transcranial Doppler (TCD) monitoring. METHODS: ECMO was conducted on 14 sheep. TCD monitoring was conducted at different time-points to examine the bilateral cerebral blood flow velocity (CBFV), including systolic peak flow velocity (Vs), end-diastolic flow velocity (Vd), mean flow velocity (Vm), and pulse index (PI, Vs-Vd/Vm). The general condition, blood pressure, heart rate, and temperature were observed. Two weeks after, the surviving 9 sheep were killed, their brains were taken out and morphological and pathological examinations were done. RESULTS: Anesthesia showed little effect on CBFV. After the ligation of carotid artery and vein, the CBFV pattern only changed slightly, Vs and PI decreased, and Vm and Vd remained almost unchanged. During ECMO with either greater or smaller volume, especially the former, the CBFV pattern showed a non-pulse waves, Vs markedly decreased with a value hardly different from that of Vd, Vs, Vd, and Vm were 62%, 75%, and 69% of the values in normal condition. The pre-ECMO CBFV was not significantly different from those examined any day after ECMO. The differences of pH and PO(2) in femoral artery during different stages were insignificant. PaCO(2) during ECMO with great flow volume and during mechanical ventilation after ECMO was significantly lower than that before ECMO (P < 0.05). The MABP of femoral artery during ECMO with small flow volume was significantly lower than that before ECMO (P < 0.05). Vm was positively correlated with MABP and not correlated with heart rate and temperature. The gross observation of the sheep's brain was normal. Small foci of malacia were seen in the brains of 4 sheep. CONCLUSION: Hemodynamics changes remarkably during ECMO. TCD monitors the CBFV continuously during ECMO, thus helping maintaining the stability of CBFV and protect the brain. PMID- 12895343 TI - [Single-lung transplantation for end-stage emphysema]. AB - OBJECTIVE: To evaluate operative technique, patient selection and perioperative management of single-lung transplantation for a patients with end-stage emphysema. METHODS: A 56-year-old patient with end-stage emphysema underwent left lung transplantation on September 28, 2002. The surgical technique used was similar to that mentioned in the literature. The donor lung was perfused by LPD solution with a cold ischemic time of 260 minutes. Cardiopulmonary bypass was not performed. RESULTS: The patient weaned from a ventilator at the 93th hour after operation. Immunosuppressants included cyclosporine, mycophenolate mofetil and corticosteroid. Acute rejection occurred on the ninth day after operation and was cured by bolus methylprednisolone given intravenously. Lung function was improved significantly and the patient was discharged from the hospital on the 47th day after operation. CONCLUSION: Single-lung transplantation for patients with end stage emphysema is effective for long-term improvement of pulmonary function. PMID- 12895344 TI - [Overexpression of COX-2 and its clinical significance in non-small cell lung cancer]. AB - OBJECTIVE: To study the relations among COX-2 expression in non-small cell lung cancer (NSCLC), its clinical characteristics and brognosis. METHODS: Immunostaining was performed with COX-2 antibody to the surgically resected tissue samples from 79 patients with NSCLC. Vessel epithelium cell COX-2 expression was taken as positive control. RESULTS: The positive rate of adenocarcinoma and squamous cell carcinoma was 85% and 57%, respectively (P = 0.013). COX-2 expression was associated with the extent of adenocarcinoma differentiation, tumor size, and TNM period, but not with the extent of squamous cell carcinoma differentiation. In the COX-2 positive group, the 5-year survival rate and a median survival time were 27.1% and 53 months; however in the negative group they were 52.0% and 61 months (P = 0.029). CONCLUSION: Invasive development of NSCLC is related to inceased expression of COX-2. COX-2 overexpression may be one of the risky factors for the prognosis of NSCLC. PMID- 12895345 TI - [Clinical significance of CEA mRNA detection in pleural and peritoneal effusions by real-time fluorescent quantitative RT-PCR]. AB - OBJECTIVES: To detect CEA mRNA levels in benign and malignant pleural and peritoneal effusions and evaluate their clinical significance. METHODS: Samples of pleural and peritoneal effusions from 58 patients with malignant diseases and 76 patients with benign diseases were collected and total RNAs were prepared and subjected to real-time fluorescent quantitative RT-PCR to determine the CEA mRNA levels in these samples. The positive rate of this examination was compared with that of shed cell pathological examination. RESULTS: Nineteen samples (32.8%) of pleural and peritoneal effusions from the 58 patients with malignant diseases showed positive results in shed cell examination, while the number of CEA mRNA >1 CN was 46 (79.3%) (chi(2) = 21.81, P = 0.000). Nineteen samples of pleural and peritoneal effusions from the 76 patients with benign diseases showed CEA mRNA > 1 CN (25.0%), which was significantly different from that of the patients with malignant diseases (chi(2) = 38.85, P = 0.000). CONCLUSION: CEA mRNA levels in pleural and peritoneal effusions can be quantified by real-time fluorescent quantitative PCR, which is more sensitive than shed cell pathological examination. This technique is helpful in discrimination of benign and malignant pleural and peritoneal effusions. PMID- 12895346 TI - [Endoscope-assisted supraorbital keyhole approach for excision of suprasellar region tumor]. AB - OBJECTIVE: To study endoscope-assisted microsurgical technique with supraorbital keyhole approach for the treatment of suprasellar region tumor so as to maximize tumor removal and minimize operative-trauma. METHODS: According to high resolution CT and MR images before operation, individual operation schemes were worked out. Skin incision was made in the eyebrow, and the diameter of supraorbital craniotomy was approximately 2 cm. Endoscope-assisted microsurgical technique was used to resect lesions in 16 consecutive patients. RESULTS: Total Tumors were removed in all of the 16 patients via the supraorbital keyhole approach. No postoperative complications occurred. CONCLUSIONS: Supraorbital keyhole approach may diminish tissue injury considerably and has proven to provide sufficient operating space in the suprasellar region for tumor removal. PMID- 12895347 TI - [The retrospective analysis of HBV and HCV infection in cholangiocarcinoma]. AB - OBJECTIVE: In order to study the diagnosis and treatment of HBV and HCV infection. METHODS: We retrospectively analysed clinical data of 680 patients with cholangiocarcinoma from 1995 to 2001 and stated by SPSS software. RESULTS: (1) The fastigium of cholangiocarcinoma was 60 - 65 years old. The incidence of cholangiocarcinoma was higher in aged males and the sex ratio (male:female) was 1.36:1. (2) The proximal cholangiocarcinoma was most (41.6%) and distant cholangiocarcinoma was secondly (28.7%). (3) Most patients of cholangiocarcinoma were late. The resection rate was low and the rate of radical operation was 21.6% (147/680). (4) The incidence of proximal cholangiocarcinoma was higher in the positive Serologic marks for HBV and HCV and course of diseases was short. Moreover, the pathology of. positive Serologic marks for HBV and HCV trended to low-differentiation and invasion, metastasis and the resection rate was lower. CONCLUSIONS: Cholangiocarcinoma is common in the aged males. The infection of HB(C)V and hilar cholangiocarcinoma are correlated and incline to the proximal bile duct. The hilar cholangiocarcinoma infected HB(C)V may have higher malignant degree in biological characteristics and more badly prognosis. PMID- 12895348 TI - [Integrative therapy for postoperative intractable enterococcus faecium infections]. AB - OBJECTIVE: To assess the treatment of patients with infection of enterococcus faecium after surgery who failed to respond to antibiotics. METHODS: Five patients after surgery were proved to have Enterococcus faecium infection by bacterial culture. They were treated by sensitive antibiotics but failed. Comprehensive treatment prescribed included immunoenhancements, enteral nutrition, and traditional Chinese medicines. RESULTS: Four patients were discharged from the hospital after recovery, and was cared else where after 1 month treatment. CONCLUSION: Comprehensive treatment is a better way to treat patients with refractory enterococcus faecium infection after surgery. PMID- 12895349 TI - [Albumin kinetics in patients with severe sepsis]. AB - OBJECTIVE: To explore the mechanism of hypoalbuminemia in patients with severe sepsis. METHODS: I(125)-labeled albumin was administered intravenously to 10 health volunteers and 10 patients with severe sepsis. Blood samples were taken at 0, 1, 2, 4, 8, 12, 24 hours and 2, 3, 4, 5, 6, 7, 9, 11, 13, 15, 18, 22, 25 days for the measurement of the dose of gamma-radiation and the curve of concentration and time. Then the half-life time (t(1/2)), apparent volume of distribution (V(d)) and transportation rate (K(12)) from center compartment to side compartment of albumin were calculated. RESULTS: The half-life time in septic group was obviously shorter than that in control group (8.2 +/- 1.4 vs. 12.5 +/- 1.7, P < 0.01). The transportation rate in the septic group was higher than that in the control group [(4.4 +/- 1.9) x 10(-2)/h vs. (2.4 +/- 0.6) x 10(-2)/h, P < 0.05]. There was no significant difference in apparent volume of distribution between the two groups. CONCLUSIONS: In patients with severe sepsis, the distribution rate of albumin from vessel to tissue was obviously increased and the decomposition rate of albumin was markedly improved. PMID- 12895350 TI - [Expression of the cyclooxygenase-2 gene in human breast carcinoma]. AB - OBJECTIVE: To study the expression of the cyclooxygenase-2 (COX-2) gene in breast cancer in contrast to that of normal breast tissues or benign breast tumors and its significance in the carcinogenesis and development of breast cancer. METHODS: With reference to the expression of the beta-actin gene, the expression of COX-2 mRNA was examined in cancerous tissues and adjacent normal breast tissue from 30 patients and benign breast tumors from 15 patients by reverse transcription polymerase chain reaction (RT-PCR). Quantitation of relative band densities was performed using densitometry-scanning software. Estrogen receptors of 30 breast cancers were investigated by immunohistochemistry. RESULTS: Enhanced expression of COX-2 was observed in ninety percent of cancers tissue with a range of 0.05 - 0.91 (median 0.53). Rare cases showed significant COX-2 expression in normal breast tissues with a range of 0 - 0.09 (median 0). In part of benign breast tumors, COX-2 expressions were obviously elevated with a range of 0 - 0.68 (median 0.07). The difference of expression of COX-2 mRNA among breast cancers, normal breast tissues, mastopathy or fibroadenomas was significant (rank-sum test, P < 0.05) and the difference of that between estrogen receptor negative and positive was also observed (rank-sum test, P < 0.01). CONCLUSION: The level of expression of COX-2 mRNA is obviously higher in breast cancer tissue than in normal breast tissue, mastopathy or fibroadenomas. The expression of COX-2 in hormone-dependent breast cancer is higher than that in hormone-independent breast cancer. The overexpression of COX-2 may play a crucial role in the carcinogenesis and development of cancer in patients with breast carcinoma. PMID- 12895351 TI - [Detection of serum levels of MMP-9 and VEGF in patients with bladder cancer]. AB - OBJECTIVE: To detect serum levels of MMP-9 and VEGF in patients with bladder cancer. METHODS: Serum levels of MMP-9 and VEGF in 58 patients with bladder cancer and 45 healthy controls were measured by sandwich-ELISA. RESULTS: Serum levels of MMP-9 and VEGF (737.12 micro g/L and 1148.88 ng/L) were significantly higher in the cancer patients than those of controls (423.51 micro g/L and 846.96 ng/L, P < 0.01). The serum levels were associated with tumor stage and grade. In patients with invasive cancer, the levels were significantly higher than those of superficial cancer (P < 0.01). Patients with distant metastasis had significantly higher levels of MMP-9 and VEGF than those with localized invasion (P < 0.01). But there was no significant difference between patients with superficial cancer and controls. Patients with G(3) tumors had significantly higher levels of MMP-9 and VEGF than those of patients with G(1) and G(2) tumors (P < 0.01). CONCLUSIONS: Elevated MMP-9 and VEGF levels are associated with a high stage and grade of bladder cancer and they may serve as markers of tumor progression in the future. PMID- 12895352 TI - [Diagnosis and treatment of seminal vesicle cyst]. AB - OBJECTIVES: To deepen the understanding of patients with seminal vesicle cyst for correct diagnosis and treatment. METHODS: Sixteen patients with seminal vesicle cysts were treated in the period of January 1980-May 2002. Their symptoms, diagnostic results, treatment and outcomes were analyzed retrospectively. The mean age of these patients at diagnosis was 31 years (range 19 - 43). Two patients were associated with ipsilateral renal agenesis. Symptoms included hematospermia in 12 (75%) patients, urinary frequency in 8 (50%), hematuria after ejaculation in 6 (27.5%), perineal malaise in 6 (27.5%), infertility in 3 (13.7%), pain after ejaculation in 3 (13.7%), scrotal pain in 2 (12.5%) and dysuria in 1 (6.3%). Cyst was palpable in 81.3% patients on digital rectal examination. All patients underwent intravenous urography and cystoscopy. Others received ultrasonography, CT scanning, MRI, and vasovesiculography. The size of masses ranged from 3.8 cm x 3.0 cm x 2.6 cm to 9.6 cm x 5.2 cm x 5.0 cm. Final open surgery consisted of vesiculectomy (4 patients) and partial vesiculectomy (12). RESULTS: Postoperative course was uneventful except in 1 patient with epididymitis. All patients were free of symptoms after open surgery. CONCLUSIONS: Seminal vesicle cysts are rare but should be considered in men with hematospermia and otherwise inexplicable bladder irritation symptoms, perineal discomfort or other genitourinary complaints of unknown etiology. Diagnosis consists of digital rectal examination, transrectal and abdominal ultrasonography, CT scan or MRI. Vesiculectomy and partial vesiculectomy give excellent results. PMID- 12895353 TI - [Selection of dorsal or volar internal fixation for unstable distal radius fractures]. AB - OBJECTIVE: To assess the results of T-plate in the treatment of unstable distal radius fractures. METHODS: Dorsal or volar T-plate fixation was used for unstable distal radius fractures according to different types of fracture. For comminuted fractures with severe cortex destruction and insufficient buttress, artificial bone graft or autograft was adopted to restore local stability. RESULTS: Satisfactory reduction was achieved after surgery. Functional rehabilitation under surveillance began from the 2nd or 3rd post-operative day. The overall excellent or good recovery rate was 90.77% after a mean follow-up of 29.63 months. For the 55 sides of volar fixation (30 sides with a dorsal displacement fragment), excellent or good recovery rate was 92.72%. Dorsal fixation was used in delayed cases (> 6 weeks) or certain fracture patterns unsuitable for volar approach. The excellent on good recovery rate was 77.77%. CONCLUSIONS: Volar approach is an ideal option not only for distal radius fracture with a volar displaced fragment but also for dorsal displaced fractures (Colles' fracture). The advantages of this approach include flat cortex for easy plate fixation, better tension band effect, less soft tissue destruction, leaving dorsal soft tissue hinge intact, and avoiding bone graft leakage. Satisfactory results can be obtained in those patients with unstable distal radius fracture through volar approach plate fixation, especially in those with fresh fractures. If possible, volar approach can also be used in delayed fractures within 6 weeks. The results of dorsal fixation is a little bit inferior. PMID- 12895354 TI - [Application of Ilizarov technique in salvage ankle arthrodesis]. AB - OBJECTIVE: To introduce the application of Ilizarov technique in ankle fusion with serious pathology and failed ankle arthrodesis. METHOD: The medical records and images of 9 cases of complex ankle fusion using Ilizarov external fixator were analyzed. Among the 9 cases, 6 received revisional ankle fusion, and 3 primary fusion for septic arthritis. All the cases were followed up for an average of 18.1 months. RESULTS: Six cases showed good results, i.e, no or mild pain, occasional limp and stable fusion and 3 cases fair results, i.e, moderate pain, persistent limp or occupational restriction, and stable fusion. CONCLUSION: Ilizarov technique is one of the effective method for the treatment of complex ankle arthrodesis. PMID- 12895355 TI - [Construction of eucaryotic expression plasmid carrying the BMP7 gene and expression in mesenchymal stem cells]. AB - OBJECTIVES: To construct an eucaryotic expression plasmid carrying the BMP7 gene and express in MSCs. METHODS: The BMP7 gene was cloned into the eucaryotic expression vector pcDNA3.1. At the same time, mesenchymal stem cells (MSCs) were isolated and cultured in vitro. The plasmid carrying the BMP7 gene was transfected into MSCs. RESULTS: PCR and digesting demonstrated that the eucaryotic expression plasmid -pcDNA-BMP7 was obtained. RT-PCR and immunohistochemical methods showed that the BMP7 gene was expressed in MSCs. CONCLUSION: Construction of an eucaryotic expression plasmid carrying BMP7 gene and expression in MSCs provide a sound basis for gene therapy using the BMP7 gene and the ideal seeds for tissue engineering. PMID- 12895356 TI - [Influence of combined cyclosporine A and tacrolimus with 5-fluorouracil on hepatocellular carcinoma rats]. AB - OBJECTIVE: To investigate the effect of combined CsA and FK506 with 5-FU on hepatocellular carcinoma rats. METHODS: A syngeneic rat model of hepatocellular carcinoma was used. Control group (A) underwent 4 ml 5% GS. Treatment group was divided into 3 groups namely, group B: only 5-FU and 5% GS; group C: 5-FU, CsA and 5% GS; group D: 5-FU, FK506 and 5%GS. Cell cycle, apoptosis, necrosis and mitochondrial transmembrane potential were measured by flow cytometry, laser scanning confocal microscopy, and electron transmission microscopy. Statistical analysis was performed by SPSS 10.0 for Windows software. Statistical comparisons were made with ANOVA followed by Dunnett's T3 or LSD test. RESULTS: Compared to the control group, the percentage of apoptotic cells including trifle necrotic cells was significantly higher, and among the treatment group, group D was the highest, and group C was higher than group B. In the treatment group, cell cycle of hepatoma cells was mainly arrested at S phase, but in group D, G0/G1 phase cells were significantly decreased and S phase cells significantly increased. Compared to the control group, mitochondrial transmembrane potential was significantly decreased in the treatment group, among with, group B was the lowest, group C was higher than group D. Morphological changes demonstrated by electron microscopy included dispersed nuclear chromatin, loss of nucleoli, membrane bleeding, cell shrinkage, typical apoptotic bodies and marked swelling of mitochondria in the treatment group. In the control group, however, they were characterized by normal cell ultrastructure. CONCLUSIONS: The present study reveals that 5-FU combined with CsA or FK506 demonstrated a synergistic effect on hepatocellular carcinoma rats. For FK506, the powerful mutual effect is related to the increase of tumor cell's quantity in S phase. Both CsA and FK506 can provide protection on mitochondrial transmembrane potential reduction against hepatoma cells damage from 5-FU. PMID- 12895358 TI - [Wound healing after pancreaticojejunostomy in piglets: a comparison between two anastomotic methods]. AB - OBJECTIVE: To evaluate wound healing after types of pancreaticojejunostomy. METHODS: After resection of the pancreatic head, 38 domestic piglets were divided into two groups according to the types of anastomoses: group I: binding pancreaticojejunostomy, a new technique designed and advocated by professor Peng Shuyou; group II: end-to-end pancreaticojejunal invagination. Anastomotic strength in vivo and histopathological findings were assessed on operative day and postoperative day 5 and 10. RESULTS: Bursting pressure was 139.7 +/- 8.0, 178.7 +/- 9.7 and 268.8 +/- 12.8 mm Hg in group I on day 0, 5 and 10, whereas 67.3 +/- 7.9, 96.2 +/- 10.4 and 130.6 +/- 9.3 mm Hg in group II. The gain on day 0 to 5 and 5 to 10 was 27.9% and 50.5% in group I and 42.9% and 35.7% in group II, respectively. A significant difference was observed between group I and group II, and between 5 and 10 day after anastomoses (P < 0.01). Breaking strength was 4.5 +/- 0.4, 6.6 +/- 0.4 and 10.0 +/- 0.6 N in group I on day 0, 5 and 10 and 4.6 +/- 0.6, 5.8 +/- 0.5 and 7.1 +/- 0.6 N in group II. Although a similar value was shown in both types of anastomoses on day 0, a rapider gain was demonstrated on day 0 to 5 and 5 to 10 in group I (44.8% and 52.9%) than in group II (25.4% and 22.0%). A significant difference was found on day 5 and 10 between the two types of anastomoses (P < 0.05 and P < 0.01). Anastomotic site was well repaired by connective tissue and the cut surface of pancreatic stump was covered by mucosal epithelium in group I on day 10, but the cut surface was incompletely repaired by granulation tissue and no, regeneration of the epithelium was found in group II. CONCLUSION: Anastomotic strength of binding pancreaticojejunostomy was stronger than end-to-end pancreaticojejunal invagination and the healing was better and rapid. PMID- 12895357 TI - [Interleukin-12 enhanced tumor necrosis factor related apoptosis-inducing ligand TRAIL-induced apoptosis in human hepatocellular carcinoma by inhibiting expression of survivin]. AB - OBJECTIVE: To investigate therapeutic potential of TRAIL in hepatocellular carcinoma (HCC) and the mechanism of sTRAIL resistance and to reverse the resistance to sTRAIL-inducing apoptosis. METHODS: The expression profiles of TRAILR were determined 60 HCC samples, in 20 normal liver tissues and 2 HCC cell lines HepG2 and SMMC-7721 by in situ hybridization. Cellular effects of sTRAIL in promoting apoptosis on HCC cell lines HepG2 and SMMC-7721 were analyzed after exposure to recombinant protein and after transfection with a cDNA expression construct. In vivo effects of sTRAIL on tumor growth were investigated using a nude mice HCC model of hepG2. Furthermore, the expression of survivin in HCC was detected, and treatment with antisence oligonucleotide was accepted. Finally, therapeutic effect on HCC by combining sTRAIL and interleukin-12 (IL-12) was detected. RESULTS: Both DR4 and DR5 were present in all HCC tissues as well as normal hepatic tissues. In contrast, 54 HCC tissues did not express DcR1 and 25 did not express DcR2. But both DcR were detectable in all of the normal liver tissues. The expression patterns of DR and DcR in HCC samples were quite different from those in normal tissue. DR5, DR4, and DcR2 expressed in both cell lines, while no DcR1 expression was detected. Recombinant sTRAIL alone was found to have a slight activity as it killed a maximum of 15% of HCC cells within 24 h while killing over 70% of Jurkat cells. In vivo administration of the TRAIL gene couldn't inhibit tumor growth in a nude mice HCC model. Mostly, HCC tissue and both HCC cell lines expressed survivin, whereas normal liver tissue did not express survivin. Treatment with antisence oligonucleotide enhanced sTRAIL inducing apoptosis. IL-12 significantly augmented sTRAIL-inducing apoptosis and inhibited survivin expression. CONCLUSIONS: HCC cells are insensitive towards TRAIL-mediated apoptosis. Survivin may play a role in resistance to TRAIL-induced apoptosis in HCC, and antisence oligonucleotide could partly reverse the resistance to TRAIL-inducing apoptosis. IL-12 may sensitize HCC cells to TRAIL induced apoptosis by preventing survivin. Combining gene therapy strategy such as combining gene therapy of TRAIL with IL-12 may be a promising maneuver to HCC. PMID- 12895359 TI - [Study of corrected visual performance and vision quality]. AB - OBJECTIVE: The present study was designed to examine the visual acuity (VA) and contrast sensitivity function (CSF) under three refractive correction methods, the visual performance and vision quality under different correction methods were compared and analyzed. METHODS: Twenty three college students with myopia (41 eyes, myopia range: -1.25 D approximately 9.25 D; astigmatism < 0.50 D) were enrolled and fully corrected by three different methods: 1) spectacles with resin lens material (SP); 2) soft contact lens (SCL); 3) rigid gas-permeable lens (RGP). The distant and near LogMar VA and CSF were examined. To avoid the subjective bias, the correction procedure was randomly selected and the VA chart and CSF chart used had the same pattern but different letters' organization or directions. RESULTS: The best corrected distant VA was significantly higher in RGP wearing eyes than those in the Soft-CL and SP wearing eyes (P < 0.01 and P < 0.05, respectively). No statistically significant difference was detected between the SP and SCL wearing eyes. There was no significant difference in the near VA among these three conditions, although the near VA in RGP group was slightly better than that in the SCL condition. There was no significant difference in CSF value at low spatial frequencies (1.5 cpd and 3 cpd) among these three conditions. However, at middle and high spatial frequencies (6 cpd, 12 cpd and 18 cpd), the RGP curve was the highest and followed by the SCL and SP curves. CONCLUSION: RGP correction offers the best visual acuity quality for myopia subjects. The CSF assessment can be used as an indirect method to evaluate the quality of corrected retinal image. PMID- 12895360 TI - [Influence of corneal refractive surgery on wavefront aberrations of human eye]. AB - OBJECTIVE: To analyze the aberration of human eyes after LASIK by using wavefront analyzer, to evaluate the eye aberrations by using the large spot in refractive surgery, and to compare the aberrations in different pupil zones. METHODS: This is a prospective clinical study. Preoperatively and postoperatively, uncorrected visual acuity (UCVA), best corrected visual acuity (BCVA), spherical equivalent (SE) and aberrations were evaluated on each patient. The patients were divided into three different groups according to the SE diopter before the surgery. We used the SAS and SPSS10.0 statistic software to analyze the data. RESULTS: Preoperatively, BCVA was 4.98 +/- 0.07. Ten days after surgery, UCVA and BCVA were 4.92 +/- 0.10 and 4.95 +/- 0.09, respectively. One month after surgery, UCVA and BCVA were 4.96 +/- 0.09 and 4.99 +/- 0.09, respectively. Over time, visual acuity improved with Postoperatively, parts of the eye's BCVA were better than preoperatively. Ten days after LASIK, the SE of three groups were hyperopic. But with time, SE changes to myopia. In the Allegretto exam, high order aberrations increased immediately after surgery in each group (P < 0.05), but decreased with time (P > 0.05) and did not return to preoperative values at one month. Preoperatively, the total aberrations (RMSg) were larger than postoperatively. The aberrations especially spherical are larger in the 6.5 mm pupil zone than in the 4 mm pupil zone. Preoperatively, there were no significant differences of high order aberrations between the three groups (P > 0.05). But after surgery, there were significant differences (P < 0.05). CONCLUSIONS: Initially, high order aberrations increased after LASIK, but decreased over time. Preoperatively, there were no significant differences of aberrations between the three groups, but after surgery, significant differences existed. The aberrations especially spherical were larger in the 6.5 mm pupil zone than in the 4 mm pupil zone. Allegretto wave-front analyzer can be used for low to middle myopia. PMID- 12895361 TI - [Topographical evaluation on decentration of orthokeratology lenses]. AB - OBJECTIVE: To evaluate the degree and correlative factors of decentration of orthokeratology lenses and its effect on the visual function. METHODS: Two different kinds of orthokeratology lenses were fitted to 270 eyes of 135 patients [initial mean refractive error: (-3.98 +/- 1.51) D]. Humphery Instruments ATLAS 8.0 was used for the computer-assisted analysis of corneal differential topographical maps. The examination of corneal topography was proceeded on the patients before the fitting of orthokeratology lenses and 6-month later. The distance from center of optic zone to apex of the cornea was measured as the value of decentration of orthokeratology lenses. The factors influenced the value of decentration were analyzed, including the initial refraction error, astigmatism, keratometry values, corneal eccentricity, and the diameter of the lens. The complaints of patients were recorded. Questionnaires, involving the symptoms of monocular diplopia and glare, were used to evaluate the effects of decentration of orthokeratology lenses on the visual function. RESULTS: The mean distance of decentration was (0.49 +/- 0.34) mm after one night fitting, the mean distance of decentration after follow-up for 1 month, 3 months and 6 months was (0.57 +/- 0.41) mm, (0.55 +/- 0.48) mm and (0.59 +/- 0.39) mm, respectively. After one month, the distance of decentration was less than 0.5 mm in 51.1% eyes, 0.5 - 1.0 mm in 35.6% eyes and more than 1.0 mm in 13.3% eyes. The direction of decentration in eyes with more than 0.50 mm decentration was mainly in the temporal side (48.5%). Patients with greater initial astigmatism and smaller diameter of lens showed greater distance of decentration (P < 0.05). There was no statistically significant difference in the distance of decentration between two groups with different corneal eccentricities and keratometry values (P > 0.05). The distance of decentration was greater in patients with monocular diplopia and glare. CONCLUSIONS: The degree of decentration of orthokeratology depends on the degree of initial refractive error, astigmatism and the design of orthokeratology lenses. The degree of decentration can influence the visual function. PMID- 12895362 TI - [Clinical research of the implantation of phakic anterior chamber intraocular lens for high myopia]. AB - OBJECTIVE: To investigate the efficacy and safety of the implantation of phakic anterior chamber intraocular lens (IOL) for high myopia. METHODS: A consecutive group of 73 eyes in 41 patients with -7.00 to -30.00 diopters (D) of myopia were implanted. RESULTS: All of 73 eyes were implanted successfully and have been followed-up for 3 m (months). The uncorrected visual acuity was from FC/33 cm to 0.2 pre-operatively and 0.1 to 1.0 3 m post-operatively. The best corrected visual acuity (BCVA) was from 0.05 to 1.0 pre-operatively and 0.1 to 1.0 3 m post operatively. The post-operative BCVA of every eye was not worse than the pre operative one. The refractive diopters were from -7.00 to -30.00 D pre operatively and -6.00 to +2.50 D 3 m post-operatively. There were no significant differences between pre- and 3 m post-operative mean corneal astigmatism (t = 1.751, P = 0.082) and mean intraocular pressure (IOP) (t = 1.181, P = 0.240), respectively. The mean counts of endothelial cells was (2 680 +/- 538)/mm(2) pre operatively and (2 514 +/- 420)/mm(2) 3 m post-operatively. There was no significant difference (t = 1.182, P = 0.242) though it decreased 6.19%. No severe complications occurred. CONCLUSIONS: The implantation of phakic anterior chamber IOL for high myopia is predictable, reversible and controllable with simple manipulation. No severe complication occurred in 3 m post-operatively, and long-time follow-up is still required. PMID- 12895363 TI - [Studies of retinal dopamine transporter in experimental myopia]. AB - OBJECTIVE: To investigate the distribution of dopamine transporter (DAT) in the chicken eye and to explore the role of DAT in the occurrence of experimental myopia. METHODS: Thirty 2-day-old chickens were divided into four groups. Chicken eyes were fitted with lenses of -10 D (Group 1), -20 D (Group 2) and translucent goggles (Group 3) unilaterally. Chickens without any treatments were used as the control (Group 4). The refraction and the axial eye length of all chickens were measured after 3 weeks, then, all chickens were given an intramuscular injection of (125)I-beta-CIT [2beta-carbomethoxy-3beta-(4-iodophenyl) tropane] and sacrificed two hours after injection. Retinal pigment epithelium (RPE) and neural retina were dissected from the eye as a whole layer in Groups 1 and 4; and dissected separately in Groups 2 and 3. Radioactive DAT from each specimen was assayed by gamma-counter. RESULTS: In Groups 2 and 3, the radioactive DAT value in the RPE from the experimental eyes was significantly greater than that in the neural retina and also greater than those in the RPE from the control eyes (P < 0.01). In Groups 1, 2, and 3, the radioactive DAT value in the whole retina or RPE from the experimental eyes was significantly greater than those from the control eyes (P < 0.01). CONCLUSIONS: Retinal DAT is located mainly in the RPE and may be involved in the occurrence of both lens-induced myopia and form deprivation myopia. These methods may provide a new approach for further studying the role of dopamine system in the occurrence of experimental myopia. PMID- 12895364 TI - [Effects of pirenzepine on lens-induced myopia in the guinea-pig]. AB - OBJECTIVE: To determine the efficacy of the M(1)-selective muscarinic antagonist, pirenzepine, in preventing lens-induced myopia in the guinea-pig and to study the mechanism and the possibility of treatment of myopia with pirenzepine. METHODS: Fifteen 4-week-old guinea-pigs were monocularly fitted with -10.00 D lenses for a period of 11 days. In Group I (n = 7), both eyes received topical administration of 0.24% saline vehicle as the controls. In Group II (n = 8), the lens-fitted eyes were topically treated with 10% pirenzepine, while the other eyes received the vehicle control. Ocular refraction and biometric measurements were collected on the first and the 11th days. All eyes were finally enucleated for histopathological examination to evaluate the possible toxic effects of pirenzepine. RESULTS: In Group I, 11 days of lens-fitting produced -2.45 D myopia (t = 3.141, P < 0.05) with 0.05 mm elongation of axial dimension (t = 2.500, P < 0.05) as compared to the contralateral eyes. There were no significantly differences of refractive error and axial dimensions between the experimental eyes and the controls in Group II. Histological examinations revealed no obviously toxic effects in the pirenzepine-treated eyes. CONCLUSIONS: Topical administration of the M(1)-selective muscarinic antagonist, pirenzepine, prevents lens-induced experimental myopia in guinea-pig by inhibiting the elongation of axial dimension with no obvious damage to the ocular tissues. PMID- 12895365 TI - [Exogenous advanced glycosylation end products induce diabetes-like vascular dysfunction in normal rats: a factor for occurrence of diabetic retinopathy]. AB - OBJECTIVE: To disclose the relationship between the deposition of advanced glycosylation end products (AGE) in the retinal vascular tissues and damage of retinal vessels in diabetic retinopathy. METHODS: Sixteen SD rats aged 2 months were divided into 4 groups, with 4 rats in each group. Rats in normal group received no treatment. Diabetes was induced by AGE in the diabetes group. Rat serum albumin (RSA, 40 mg/kg weight) was administered daily to healthy non diabetic rats through tail veins for 2 weeks (RSA group). AGE-modified RSA was injected to rats in another group at the same route and dosage (AGE-RSA group). The number of pericytes in retinal capillary vessels was counted 2 weeks later. RESULTS: After two weeks continuous AGE treatment, the average amount of pericytes of capillary vessel per 10 microscope visual field (x 100 magnification) in AGE group (4.31 +/- 0.34) was significantly less than that of RSA group (5.80 +/- 0.48) (P < 0.01). Meanwhile, in the AGE-RSA group, AGE were identified in the retinal vascular tissues by immunohistochemical staining. CONCLUSION: Injection of exogenous AGE into healthy rats induces vascular changes resembling those find in the diabetic retinopathy. AGE might be one of the independent pathogenic factors in the occurrence of diabetic retinopathy. PMID- 12895366 TI - [Induced differentiation of neural stem cells from subependymal zone into retinal cells in vitro]. AB - OBJECTIVE: To study the cultivation and differentiation of neural stem cells (NSC) in vitro and observe the effects of retinal cell-conditioned medium on the differentiation of NSC. METHOD: Cells from subependymal zone of postnatal 0 - 3 days SD rat were isolated and cultured in vitro. Retinal cells of SD rat were cultured simultaneously. The supernatants (conditioned medium, SDR-CM) of cultured retinal cells were collected and used for the cultivation of the NSC from subependymal zone. Immunofluorescence method and anti-Thy1.1 antibodies were used to identify the cells derived from the NSC. RESULTS: Cultured NSC grew well in the serum-free culture medium. Cultured subependymal cells in the SDR-CM could be differentiated to the retinal cells. Some cells were stained positively with anti-Thy1.1 antibodies. CONCLUSION: These results showed that cultured NSC could survive well in vitro. SDR-CM can induce the NSC to differentiate to the retinal cells. The present study was designed to simulate the microenvironment of the eyes and to induce the differentiation of the NSC. Our in vitro model system may provide theoretical basis for the intraocular transplantation of retinal neurons. PMID- 12895367 TI - [A quantitative study on early changes in rabbit lens capsule epithelium induced by low power density microwave radiation]. AB - OBJECTIVE: To test the safety of guidelines for limiting exposure to low power level microwave radiation and to try to explore the mechanisms of early damages induced by such radiation in rabbit lens capsule epithelial cells, using flow cytometry as a quantitative method of evaluation of these damages. METHODS: The rabbit eyes were exposed to 5 mW/cm(2) (group I) and 10 mW/cm(2) (group II) power densities of microwave for 3 h. Annexin V-PI two-color flow cytometry was used to detect the changes of rabbits lens epithelial cells, apoptosis cells and secondary necrotic cells after radiation. The other eyes served as self-controls. RESULTS: In group I, the percentage of early apoptosis cells in the experimental eyes was significantly greater than that of the controls (P < 0.01), whereas no significantly difference in the percentage of secondary necrotic cells could be observed (P > 0.05). In group II, there was no significant difference between the percentage of early apoptosis cells in experimental eyes and the controls (P > 0.05), whereas the percentage of secondary necrotic cells in the experimental eyes was significantly greater than that of the controls (P < 0.05). The percentage of secondary necrotic cells (16.2 +/- 5.6)% in group II is greater than that in group I (10.3 +/- 6.8)% (P < 0.05), though there is no significant difference in the percentage of early apoptosis cells between these two groups (P > 0.05). CONCLUSION: Low power density of microwave radiation (5 mW/cm(2) and 10 mW/cm(2)) can induce irreversible damage of rabbit lens epithelial cells in a dose-dependent manner. Therefore, the safety dosage criterion of microwave exposure needs further investigation. PMID- 12895368 TI - [Creating the model county to promote the prevention of blindness in Zhejiang Province]. AB - OBJECTIVE: To develop a system of enhancing work efficiency in prevention of blindness in Zhejiang province by creating the Model County in prevention of blindness. METHODS: Twenty counties were selected in the whole Zhejiang province to serve as the candidates of the Model County in prevention of blindness in 1996 - 1998. The activities for prevention of blindness were conducted in these counties according to the five standards of the Model County in prevention of blindness established by the National Committee for Prevention of Blindness. The Model County in prevention of blindness was awarded only after checking and accepting by the authorities in county, provincial and national levels. RESULTS: The effective way for creating the Model County in prevention was as follows: the local government put the prevention of blindness in the list of the priorities, the local committee for prevention of blindness was established, the three level network for prevention of blindness was established, the eye care workers were trained, the necessary equipment was provided. After the efforts for three years, seven counties were awarded as the Model County in prevention of blindness. In addition, Shaoxing Prefecture city became Model City in prevention of blindness. A strong effort was made to promote the activities for prevention of blindness during 1996 to 1998 in the other thirteen counties. The number of eye doctors increased from 602 into 1 850. There were two folds increase in the number of equipment availability, for example, slit lamp and ophthalmic microscope. The annual number of cataract surgeries was about 20 000. CONCLUSION: It appears to be a good approach to create the Model County to promote the prevention of blindness. PMID- 12895369 TI - [Monitoring novel ovarian carcinoma associated genes using cDNA expression microarray]. AB - OBJECTIVE: To explore the gene expression pattern of sample of human ovarian carcinoma. METHOD: The difference in gene expression between normal and neoplastic human ovarian tissues were investigated, we described the assembly and utilization of a 512 member cDNA microarray. RESULT: Thirty-seven genes expressed in ovarian cancer were screened out, 14 genes were up-regulated, 23 genes were down-regulated. CONCLUSION: cDNA microarray for analysis of gene expression pattern is an effective method to identify novel ovarian cancer associated genes. PMID- 12895370 TI - [Investigation on the gene expression pattern in cervical squamous cell carcinoma of uygur]. AB - OBJECTIVE: To investigate the differentially expressed gene in cervical squamous cell carcinoma of uygur and normal cervical tissue using gene chip. METHOD: Gene chips containing 2,048 human genes were used to investigate the gene expression pattern of 3 samples of cervical cancer. RESULT: By applying this gene chip we identified 64 differentially expressed genes in all 3 clinical cervical cancer samples, and we further identified 53 upregulated and 11 downregulated genes for functional analysis. CONCLUSION: The analysis of gene expression pattern of tumor based on gene chip can realize high-throughput screening of the genes associated with the cervical cancer. PMID- 12895371 TI - [Study of DNA microarray chip of associated genes of hydatidiform mole]. AB - OBJECTIVE: To screen genes which may associate with the development and malignant transformation of hydatidiform mole. METHOD: The differentially expressed genes were analyzed between the tissues of two cases of hydatidiform mole and the normal placental tissues of two cases of pregnancy which had almost the same pregnant ages as hydatidiform mole, using cDNA chips containing 4,096 genes. RESULTS: There were total 89 genes significantly differently expressed in all hydatidiform moles, counting for 2.2% of total genes. Compared with normal villi, 24 genes in hydatidiform mole were up-regulated and 65 genes were down-regulated. Bioinformatical analysis of genes showed genes associated with cell proliferative inhibition, such as Ras GTPase activating protein, TGF-beta IIR alpha, BTG2 were significantly down-regulated, whereas genes associated with cell proliferation, malignant transformation and tumor metastasis as thymidine kinase, ribonucleotide reductase, glucose transport protein were highly up-regulated. CONCLUSION: cDNA chip technique is one kind of very effective methods in screening associated genes in hydatidiform mole. PMID- 12895372 TI - [Effect of toxoplasma infection during pregnancy to the development of fetus and infant]. AB - OBJECTIVE: To investigate the effect of Toxoplasma (Tox) infection during pregnancy to the development of fetus and infant. METHODS: Enzyme linked immunoadsorbent assay (ELISA) was employed to detect Tox-IgM in peripheral blood of 3,908 pregnant women. To Tox-IgM positive cases, the umbilical blood of their neonates were sampled to detect Tox-IgM and abortion tissues were sampled to detect Tox-DNA with polymerase chain reaction. Infection group included 95 cases whose Tox-IgM or DNA were positive, and control group consisted of 119 cases without infection. Different supervision methods were used to periodically observe the intelligence development of infants 3 and 12 months after birth in these two groups. After initial intervention, verbal intelligence quotients (VIQs), performance intelligence quotients (PIQs) and full scale intelligence quotients (FSIQs) were assessed when the infants were 1 and 4 years old of these two groups. RESULTS: The incidences of abortion, stillbirth, pre-term delivery, FGR and malformation in infection group were 12 cases (12.6%), 5 cases (5.3%), 4 cases (4.2%), 4 cases (4.2%) and 3 cases (3.2%), while the incidences of these abnormalities in control group were 3 cases (2.7%), 1 cases (0.9%), 2 cases (1.8%), 2 cases (1.8%) and 1 cases (0.9%). Comparing with these two groups, there was significant difference (P < 0.01). Relative risk (RR) of these 5 abnormalities was 4.7, 5.9, 2.3, 2.3 and 3.6 respectively. Mental development index (MDI) and psychomotor development index (PDI) of infants in infection group were 93 +/- 13 and 101 +/- 16. They were significantly lower than that of control group, which were 107 +/- 17 and 111 +/- 13. There was significant difference between these two groups (P < 0.05). After intervention in 1 and 4 years, their VIQs, PIQs and FSIQs were apparently improved (P < 0.05). CONCLUSION: Congenital Tox infection is closely correlated with abnormal pregnancy outcomes and infantile mental retardation. Early intervention can improve intelligence development. PMID- 12895373 TI - [Maternal and perinatal prognosis affected by the time of termination of pregnancy in patients with hemolysis elevated liver enzymes and low platelet syndrome]. AB - OBJECTIVE: To investigate the optimal time of termination of pregnancy for patients with hemolysis elevated liver enzymes and low platelet (HELLP) syndrome. METHODS: 57 patients with HELLP syndrome admitted from October 1992 to September 2001 were enrolled. According to the length from the time diagnoses confirmed to the time of delivery, patients were divided into 3 groups; group I, within 24 hours, group II, 24 to 48 hours and group III, over 48 hours. Complications, maternal and perinatal mortality were analyzed retrospectively between different groups. RESULTS: Maternal and perinatal mortality were 7% and 11% in group I, 16% and 21% in group II, 64% and 73% in group III with significant differences between group III and group I or group III and group II (P < 0.05). Incidence of DIC, ARF and neonatal asphyxia was 11%, 4% and 19% in group I compared with 55%, 36% and 64% in group III, significantly higher in group III than those in group I (P < 0.05). Incidence of ARF in maternal was 4% in group I and 37% in group II with significant difference (P < 0.05). CONCLUSION: Pregnancy should be terminated as soon as possible once diagnosis of HELLP is confirmed with optimal time within 48 hours. PMID- 12895374 TI - [Kinetics of protein kinase C activities in kidney of fetal rat after intrauterine distress]. AB - OBJECTIVE: The aim of this study is to observe the kinetic changes of protein kinase C (PKC) activities in kidney of fetal rat after intrauterine distress (IUD), and to investigate the roles of PKC in the pathogenesis of kidney impair in IUD of fetal rat. METHODS: SD rats pregnant for 21 days were anesthetized intraabdominally, then one side of vessels of the two horns of uterus were occluded by arterial clamp to make models of intrauterine ischemia, hypoxia (IH) and reperfusion of fetal rat. The fetal rats were divided into IH group (ischemic and hypoxic for 15 min and 30 min, 16 fetal rats for each time point), reperfusion group (ischemic and hypoxic for 30 min, then reperfused for 15 min, 30 min, 45 min, 1 h and 2 h, respectively. 16 fetal rats for each time point) and false operation group (FOG) (18 fetal rats unclamped in the other uterus horn). The kidney tissue was homogenized first, then cell membrane protein and cytoplasmic protein were extracted. Improved Takai method was used to detect the changes of PKC activities of cell membrane and cytoplasma in kidney tissue of fetal rats. RESULTS: (1) PKC activity of cell membrane started to increase after ischemia. The PKC activity of IH group was (5.66 +/- 0.88) pmol.mg(-1).min(-1); FOG was (4.30 + 0.97) pmol.mg(-1).min(-1). The PKC activity of IH group was significantly higher than that of FOG (P < 0.01). PKC activity of cell membrane increased rapidly after reperfusion, reached a peak at 30 min, and then decreased. The PKC activity was (7.26 +/- 0.76) pmol.mg(-1).min(-1) at 15 min, (9.25 +/- 0.94) pmol.mg(-1).min(-1) at 30 min, (8.34 +/- 0.89) pmol.mg(-1).min( 1) at 45 min, (6.57 +/- 0.96) pmol.mg(-1).min(-1) at 1 h, respectively. The PKC activity of reperfusion group was significantly higher than those of FOG (P < 0.01). The PKC activity of reperfusion group was (4.64 +/- 0.96) pmol.mg(-1).min( 1) at 2 h. There was no significant difference compared with that of FOG (P > 0.05). (2) The cytoplasmic PKC activity changed in a tendency opposite to that of the cell membrane PKC activity. In IH group, the PKC activity was (9.74 +/- 1.25) pmol.mg(-1).min(-1) at 15 min, (8.47 +/- 0.84) pmol.mg(-1).min(-1) at 30 min. FOG group was (10.63 +/- 1.92) pmol.mg(-1).min(-1). Reperfusion group reached the lowest PKC activity when reperfused for 30 min, (6.60 +/- 0.94) pmol.mg(-).min( 1), with a significant difference compared with that of FOG (P < 0.01). Then the PKC activity recovered to the level of FOG when reperfused for 2 h, (9.86 +/- 1.00) pmol.mg(-1).min(-1). CONCLUSIONS: After intrauterine ischemia, hypoxia and reperfusion, PKC moves rapidly from cytoplasm to cell membrane in kidney tissue of fetal rat. PKC may be an important intracellular second messenger. It may play an important role in kidney impair in perinatal period. PMID- 12895375 TI - [The blood lead level and pregnant outcome in pregnant women with non occupational lead exposure]. AB - OBJECTIVE: The aim of this study is to find out the relations between maternal blood lead and pregnancy complications, neonatal physical and neurobehavioral development respectively. METHODS: 1,151 samples of maternal venous blood and 434 samples of umbilical cord blood were collected. Graphite stove atomic absorption spectrophotometer was used to detect the lead concentration. Neonatal physical indexes were measured within 24 hours of age, including weight, length, head circumference, chest circumference and abdomen circumference. The neurobehavioral developments were scored at 42 days of age. Meanwhile, the pregnancy complications were obtained from the medical records. RESULTS: (1) the mean lead concentration of maternal blood is 43.4 microg/L while 44.5 microg/L of first trimester, 37.4 microg/L of second trimester and 45.1 microg/L of third trimester. The mean lead concentration of cord blood is 26.4 microg/L. (2) In this study, the blood lead level is not correlated with pregnancy complications and neonatal physical and neurobehavioral development, but it is positively correlated with maternal haemoglobin concentration. CONCLUSIONS: In this study, the situation of maternal blood lead level is acceptable. At such level, the lead concentration of pregnant women blood can't influence the incidence of pregnancy complications, neonatal physical and neurobehavioral development. PMID- 12895376 TI - [Clinical application of a modified pronuclear stage scoring system following conventional in-vitro fertilization]. AB - OBJECTIVE: To assess the clinical value of pronuclear stage scoring system in human conventional in vitro fertilization-embryo transfer (IVF-ET) program. METHODS: A modified pronuclear scoring system was used to score zygotes 16 - 18 hours after insemination during conventional IVF. RESULTS: A total of 178 IVF cycles were recruited in this study. Cycles with mean pronuclear score (sum of pronuclear scores of all transferred embryos divided by total number of transferred embryos) > or = 13 had significant high pregnancy and implantation rate, as compared to those with less than 13 (49.1% and 26.5% Vs 29.4% and 15.0% respectively). Although the pregnancy, implantation rate and multiple pregnancy rate tend to increase with the increasing number of zygotes with full pronuclear score (= 15) among transferred embryos, no significant difference could be found (P > 0.05). When all embryos were analyzed, 77.2% zygotes with pronuclear score > or = 11 could be transferred or cryopreserved, while those with pronuclear score less than 11 decreased to 46.1%. CONCLUSIONS: Pronuclear stage scoring system provides additional parameter for embryo selection. However, pronuclear morphology could only partly represent embryonic viability. Combination with the scoring of both pronuclear stage and cleavage stage embryos for selecting transferred embryos is indispensable. PMID- 12895377 TI - [Expression of matrix metalloproteinases-9 and tissue inhibitor of metalloproteinases-1 mRNA in the endometrium during mid- luteal phase in women with unexplained infertility]. AB - OBJECTIVE: To investigate the expression of matrix metalloproteinases-9 (MMP-9) and tissue inhibitor of metalloproteinases-1 (TIMP-1) mRNA in the endometrium during mid-luteal phase in women with unexplained infertility and its steroidal regulation. METHODS: In situ hybridization method was utilized to detect the location of MMP-9 and TIMP-1 mRNA. Reverse transcription polymerase chain reaction (RT-PCR) method was utilized to detect and quantitate MMP-9 and TIMP-1 mRNA expression levels in the endometrium during mid-luteal phase. Serum concentration of estradiol (E(2)) and progesterone (P) were measured by radioimmunoassay. Thirty-eight patients with unexplained infertility were included as study group, and 20 women with male factor infertility or normal volunteers were selected as control group. RESULTS: MMP-9 and TIMP-1 mRNA were diffusely distributed in the cytoplasm and nuclei of glandular and stromal cells. Their staining in control group were more abundant than those in study group. Endometrial MMP-9 and TIMP-1 mRNA expression during mid-luteal phase showed by RT PCR in the study group were significantly lower than those in the control group (0.42 +/- 0.19 Vs 0.57 +/- 0.19, P < 0.05; 0.59 +/- 0.19 Vs 0.81 +/- 0.20, P < 0.01 respectively). Serum P of the study group was significantly lower than that of control group [(34 +/- 15) nmol/L Vs (53 +/- 17) nmol/L, P < 0.01]. Serum P level was positively correlated with MMP-9 mRNA only in control groups. CONCLUSIONS: MMP-9 and TIMP-1 mRNA might play an important role in the process of implantation. The decreased expression of their gene related to P level might be one of the major causes in unexplained infertility. PMID- 12895378 TI - [A clinical analysis of 146 cases of endometrial carcinoma with medical disorders]. AB - OBJECTIVE: To report the associate medical disorders of the patients with endometrial carcinoma and analyze the medical disorders' effects on the treatment and prognosis. METHOD: One hundred and forty-six cases of endometrial carcinoma associated with medical disorders who admitted to our center from July, 1971 to Oct, 2001 were analyzed retrospectively. And the other 161 cases without medical disorders who admitted to our center during the same time were chosen as control. RESULTS: Among the 146 patients with medical disorders, the ordinary disorders were hypertension in 104 patients (71.2%), diabetes mellitus in 30 patients (20.5%) and coronary heart disease in 29 patients (19.9%). Those patients with medical disorders were elder than the control (P < 0.001). The medical disorders were independent of stage, tumor cell differentiation and pathological classification (P > 0.05). The operation ratio of those patients with medical disorders were less than the control. But there was no significance (P > 0.05). The radiotherapy and chemotherapy ratio of them was significantly higher than the control (P < 0.01). Univariate analysis of the cases followed up showed that the prognosis were independent of the associated medical disorders, such as hypertension, diabetes mellitus and coronary heart disease (P > 0.05). Multivariate analysis showed that the prognosis were related to the stage and tumor cell differentiation affected the prognosis remarkable. But the prognosis was independent of age and associated medical disorders. CONCLUSIONS: The endometrial carcinoma often associates with hypertension, diabetes mellitus, coronary heart disease, and so on. The incidence rate of medical disorders increases with the aging of the patients. The medical disorders might affect the therapy to a certain extent. But they do not affect the prognosis remarkably. PMID- 12895379 TI - [Study of the therapeutic effects of costimulatory molecules B7-1 and interleukin 12 modified tumor cells vaccines on ovarian epithelial carcinoma through induction of anti-cancer immunity of the body]. AB - OBJECTIVE: To study the biological characters of tumor cell vaccines modified by interleukin 12 (IL-12) and co-stimulatory molecule B(7 - 1) and their combination therapeutic effect on rat ovarian cancer animal model. METHODS: Retroviral vector pLmB(7 - 1)SN, which expressed murine B(7 - 1) was constructed. An epithelial ovarian cancer cell line NuTu-19 was infected with pLmIL-12SN and/or pLmB(7 - 1)SN by lipofectin-mediated gene transfer system. Cell lines that could highly express either or both of these cytokines were named NuTu-19/IL-12, NuTu-19/B(7 - 1) or NuTu-19/IL-12-B(7 - 1), according to the results of enzyme-linked immunosorbent assay (ELISA) and flow cytometry (FCM). NuTu-19/Neo, the cell line that was transfected by vector pLXSN was used as a control. Various types of cytokine-modified tumor cells were injected subcutaneously into syngeneic rats Fischer 344 and their tumorigenecities were recorded. After being immunized twice by various types of mitomycin C treated gene modified tumor cells, the survival time of the intraperitoneal disseminating ovarian cancer animal model was observed, and the anti-tumor mechanisms of different gene modified tumor cell lines were discussed. RESULTS: The reconstruction of pLmB(7 - 1)SN was successful. Stable IL-12 and B(7 - 1) expression in cell lines NuTu-19/B(7 - 1), NuTu-19/IL-12 and NuTu-19/IL-12-B(7 - 1) were confirmed by ELISA and FCM. Tumorigenecities of various gene modified tumor cells decreased in syngeneic rats. The splenic lymphocytes proliferation indices (PI) in B(7 - 1) group (NuTu 19/B(7 - 1) immunized group) and IL-12 group (NuTu-19/IL-12 immunized group) were 2.4 and 4.6 (the letter P < 0.05 compared with that in control group), while PI in B(7 - 1) and IL-12 combination group (NuTu-19/IL-12-B(7 - 1) immunized group) increase to 10.2, being of significantly difference compared with those in control or B(7 - 1) or IL - 12 group (P < 0.01). More significant cytotoxic effects of cytotoxic lymphocytes (CTLs) on syngeneic tumor cells could be observed in B(7 - 1) group (15.0%) or IL-12 group (31.5%) (P < 0.05 or P < 0.01, compared with 2.6% in control group), while coexpression of IL-12 and B(7 - 1) was of great importance in enhancing CTL activity (52.4%), compared with that in control or B(7 - 1) or IL-12 group (P < 0.05, respectively). The life spans of ovarian cancer-bearing rats immunized by IL-12 (59.8 d) or B(7 - 1) (56.2 d) tumor cell vaccines were a little longer than that in control group (53.4 d), but no significant differences existed (P > 0.05), whereas in the animal models who had received IL-12 and B(7 - 1) co-immunization, prolonged survival time was showed which had statistical significance compared with that in control group (66.0 d, P < 0.05). CONCLUSION: B(7 - 1) and IL-12 modified tumor cell vaccines can induce anti-tumor immunity through stimulating lymphocytes proliferation and inducing recognition and cytotoxic effects of CTLs on tumor cells. An obvious synergistic effect exists when the two cytokines are combined. Combined immunogenic therapy with IL-12 and B(7 - 1) should prove to be a promising therapeutic strategy in ovarian cancer. PMID- 12895380 TI - [Inhibition effects of c-erbB-2 and c-raf-1 antisense oligodeoxynucleotides combined transfection on the human ovarian carcinoma transplanted subcutaneously in nude mice]. AB - OBJECTIVE: To investigate the inhibition effects of c-erbB-2 and c-raf-1 antisense oligodeoxynucleotides (ASODN) combined transfection on the human ovarian epithelial cancer transplanted subcutaneously in nude mice. METHODS: There were 7 groups: normal control group, c-erbB-2 sense observed group, c-raf-1 sense observed group, c-erbB-2 antisense observed group, c-raf-1 antisense observed group, whole dose combined group, half dose combined group. Human ovarian epithelial cancer cells SKOV3 were treated by different oligodeoxynucleotides, then transplanted subcutaneously in nude mice, respectively. The changes of tumor volume were observed and the tumor growth inhibitory rate was calculated. RESULTS: There was no difference between sense observed group and normal control group. There was a larger growth inhibitory rate in whole -dose combined group and half -dose combined group, the first time that can be detected was 13.7 days and 15.2 days, and the maximum tumor growth inhibitory rates were 61.1% and 71.3%, respectively. CONCLUSIONS: The results suggested that ASODN combined transfection can inhibit the tumorigenesis of ovarian epithelial cancer cells in nude mice, it may be a more useful gene therapy for the ovarian epithelial carcinoma. PMID- 12895381 TI - [Influences of active immunotherapy on T helper cell type 1 and 2 cytokines in women with unexplained habitual abortion]. AB - OBJECTIVE: To investigate the influences of active immunotherapy on T helper cell (Th)1/Th2 type cytokines in women with unexplained habitual abortion (UHA). METHODS: A total of 55 patients with UHA were studied, including 30 cases after active immunotherapy (AIT) and 25 cases without any therapy (NAIT). Fifteen cases of normal nonpregnant (NNP) women were selected as control group. Supernatants from trophoblast-activated peripheral blood mononuclear cells (PBMC) of the three groups were tested by enzyme-linked immunosorbent assay (ELISA) for interferon gamma (IFN-gamma), interleukin-2 (IL-2), IL-4, IL-10. RESULTS: (1) The levels of IL-2 and IFN-gamma in AIT group [(108 +/- 37) ng/L and (110 +/- 52) ng/L, respectively] were lower significantly than those in NAIT group [(223 +/- 85) ng/L and (326 +/- 92) ng/L, respectively] (P < 0.05). The levels of IL-4 and IL 10 in AIT group [(50 +/- 11) ng/L and (140 +/- 37) ng/L, respectively] were higher than those in NAIT group [(23 +/- 11) ng/L and (52 +/- 28) ng/L] +/- (P < 0.05). The levels of IL-2 and IFN-gamma in NAIT group were higher than those in NNP group [(92 +/- 32) ng/L and (102 +/- 35) ng/L] (P < 0.05). The levels of IL-4 and IL-10 in NAIT group were lower than those in NNP group [(62 +/- 21) ng/L and (150 +/- 42) ng/L] (P < 0.05). The level of every cytokine had no difference in AIT group and NNP group (P > 0.05). (2) Twenty-six women in AIT group got pregnant, but 8 women experienced pregnancy loss repeatedly whose IL-2, IFN-gamma levels were higher than those in other 18 women got successful pregnancy and IL 4, IL-10 levels lower than the latter. CONCLUSIONS: UHA patients have Th1 type immunity to trophoblast and produce high-level Th1 type cytokines which probably result in pregnancy loss. Active immunotherapy could make a shift from Th1 to Th2 immunity, thus favoring the maintenance of pregnancy. PMID- 12895382 TI - Randomized trials in oncology: a fertile ground for controversy. PMID- 12895383 TI - Rituximab and CHOP for elderly patients with diffuse large B-cell lymphoma. PMID- 12895384 TI - Rituximab for Lymphocyte-predominant Hodgkin's disease. PMID- 12895385 TI - Posttransplant lymphoproliferative disease: pathogenesis, monitoring, and therapy. AB - The spectrum of transplant-related lymphoproliferative diseases is expanding to include a variety of neoplasias that typically occur late after transplant including Epstein-Barr virus (EBV)-negative B- and T-cell lymphomas, EBV-positive T-cell lymphoma, myeloma, plasmacytoma, and Hodgkin's disease. New approaches to diagnosis and monitoring based on quantitative polymerase chain reaction for EBV DNA are being explored. What exactly is being measured (the source and character of the viral DNA) remains to be determined, as does the compartment that should be assayed (whole blood, serum, plasma, or lymphocytes). These questions not withstanding, there is an emerging consensus that these technologies will facilitate rapid diagnosis and therapeutic monitoring in the future. A myriad of therapeutic interventions are or will become available. Rituximab, alone or in addition to other therapies, promises a profound change in the landscape with regard to the treatment and perhaps the prevention of posttransplant lymphoproliferative disease. New approaches to adoptive cellular immunotherapy, including use of EBV-specific products from unrelated donors, nonspecifically activated autologous products, and genetically engineered T cells, are all being explored. PMID- 12895386 TI - New strategies in radioimmunotherapy for lymphoma. AB - Treatment options for patients with indolent non-Hodgkin's lymphoma historically involved radiation or chemotherapy. Although initial response rates are excellent, treatment is increasingly less effective with each successive relapse. The advent of immunotherapy heralds a new era for the treatment of these patients. Radioimmunotherapy adds the benefits of cytotoxic radiation to immunotherapy and represents a significant addition to the treatment armamentarium. Various antigens for lymphoma have been targeted, of which anti CD20 antibodies are the furthest in development. Ibritumomab tiuxetan (Zevalin; IDEC Pharmaceuticals, San Diego, CA), a (90)yttrium-labeled agent, and (131)iodine-labeled tositumomab (Bexxar; Corixa, Seattle, WA) are approved by the US Food and Drug Administration. Both agents have shown utility in therapy for relapsed and refractory low-grade and transformed lymphomas. This review highlights features of radioimmunotherapy that are relevant to non-Hodgkin's lymphoma, focusing on the two anti-CD20 antibodies. PMID- 12895387 TI - Simian virus 40 and its association with human lymphomas. AB - Simian virus 40 (SV40) is a potent DNA tumor virus that is known to induce cancer in laboratory animals. The neoplasias induced by SV40 in animal models are brain cancers, mesothelioma, bone cancers, and systemic lymphomas. SV40 oncogenesis is mediated primarily by the viral large tumor antigen, which inactivates the tumor suppressor proteins p53 and pRb family members. Evidence indicates that SV40 is an emergent human pathogen and that a significant excess risk of SV40 is associated with primary human brain cancers, malignant mesothelioma, bone cancers, and non-Hodgkin's lymphoma. Therefore, the major types of tumors induced by SV40 in laboratory animals are the same as those human malignancies found to contain SV40 markers. Experimental and clinical data indicate that SV40 may be functionally important in the development of some of those malignancies. Recently, the Institute of Medicine of the National Academies concluded that SV40 infections could lead to cancer in humans under natural conditions (based on moderate strength biologic evidence). This review examines the data implicating SV40 in the pathogenesis of human lymphomas and discusses future directions to define the causative role for SV40 in these malignancies. PMID- 12895388 TI - Vaccination strategies for lymphomas. AB - Vaccination strategies for lymphomas were developed along with one of the first recognized tumor-specific targets, the clonal antigen receptor, composed of unique variable regions known as idiotypes. Human clinical trials of idiotype vaccination have benefited from highly concordant animal models, leading to sequential improvements in design. Evidence of the clinical benefit of idiotype vaccines is strong but formally unproven. Significant progress has been made in our understanding of the basic mechanisms underlying the induction of immune responses, which has led to a proliferation of rationally designed immunotherapeutic strategies. Current research efforts include the development of more convenient methods to produce individual idiotype vaccines, the establishment of definitive proof for clinical efficacy, and the implementation of alternative vaccination strategies, including genetic vaccination and genetically or immunologically modified autologous tumor cells and dendritic cells. PMID- 12895389 TI - Early blast clearance by remission induction as a prognostic factor in acute myeloid leukemia. PMID- 12895390 TI - Adaptive randomization in a treatment study of patients with adverse karyotype acute myeloid leukemia. PMID- 12895391 TI - Monitoring PML-RARalpha in acute promyelocytic leukemia. AB - Acute promyelocytic leukemia (APL) is characterized by a translocation between the promyelocytic leukemia gene (PML) on chromosome 15 and the retinoic acid receptor-alpha (RARalpha) gene on chromosome 17. Reverse-transcription polymerase chain reaction (RT-PCR) amplification of PML-RARalpha messenger RNA can establish the diagnosis of APL, predict response to all-trans retinoic acid and arsenic trioxide, detect minimal residual disease, and predict relapse. Quantitative "real-time" RT-PCR techniques may improve residual disease assessment by facilitating more rapid and standardized results. APL provides a useful model in which therapy is targeted to an underlying genetic aberration and treatment is adapted based on monitoring of residual disease. PMID- 12895392 TI - Monitoring AML1-ETO and CBFbeta-MYH11 transcripts in acute myeloid leukemia. AB - The core-binding factor (CBF) leukemias comprise acute myeloid leukemia (AML) with t(8;21) and inv(16)/t(16;16), characterized by the presence of the AML1-ETO and CBFbeta-MYH11 fusion genes, respectively. These leukemia-associated genes can now be sensitively and reliably quantified by real-time reverse transcription polymerase chain reaction (RT-PCR) techniques and thus can serve as molecular targets for monitoring residual leukemia. Studies to date suggest that quantitative monitoring of minimal residual disease (MRD) in CBF-positive AML is useful in distinguishing patients at high risk of relapse from those in durable remission. Preliminary results of MRD monitoring by real-time RT-PCR in this subset of AML patients are promising and provide the basis for further evaluation by quantitative analysis in large prospective clinical trials. PMID- 12895393 TI - Monitoring of acute myeloid leukemia by flow cytometry. AB - Monitoring of minimal residual disease (MRD) becomes increasingly important in the risk-adapted management of patients with acute myeloid leukemia (AML). In selected patients with AML, multiparameter flow cytometry has shown accuracy and sensitivity in the quantification of MRD levels with independent prognostic impact. The applicability of this approach is superior to that of other methods such as quantitative polymerase chain reaction: Up to 80% of all patients can be monitored by flow cytometry. Nonetheless, significant technical advances are anticipated to extend the applicability of flow cytometry to 100% and to improve its sensitivity. Large clinical trials will determine the role of immunologic monitoring in the prognostic stratification of patients with AML. PMID- 12895394 TI - Immunologic monitoring in adults with acute lymphoblastic leukemia. AB - Investigation of minimal residual disease (MRD) by immunophenotyping and molecular techniques has proven to be a powerful approach for disease monitoring in patients with acute leukemia. Multiparameter flow cytometry, through the use of triple or quadruple marker combinations, identifies aberrant or uncommon phenotypic profiles in more than 90% of adult patients with acute lymphoblastic leukemia (ALL) at diagnosis. These profiles allow identification of residual leukemic cells in bone marrow or peripheral blood once morphologic complete remission is achieved. Until now, most immunophenotypic MRD studies in ALL have focused on children. In contrast, information on the value of MRD in adults with ALL is scanty and usually restricted to polymerase chain reaction studies. In this review, we focus on technical aspects of MRD detection by flow cytometry and on the clinical data concerning the value of immunologic MRD studies as a tool for relapse prediction in adult ALL. Although prospective studies are needed, we assert that immunophenotypic MRD studies are clinically useful. Such studies should be incorporated into the routine management of adult ALL patients for identification of those at high risk of relapse, who could benefit from new alternative therapeutic approaches, and to distinguish these patients from others who could be cured with more conventional approaches. PMID- 12895395 TI - Immunologic monitoring in chronic lymphocytic leukemia. AB - Chronic lymphocytic leukemia (CLL) is characterized by progressive defects in humoral- and cell-mediated immunity. These defects are manifested as a propensity to develop infections with encapsulated bacteria, and less frequently, with gram negative enterics. In addition, reactivation of viruses such as herpesvirus is not uncommon. Treatment of the disease further exacerbates immunosuppression by depleting immune effectors and broadening the spectrum of potentially offending pathogens. Risk of infection can potentially be reduced by administration of intravenous immunoglobulin and use of prophylactic antibiotics for individuals who are at high risk. Current work focuses on development of cellular and cytokine therapy to facilitate immune reconstitution in patients with CLL, thereby reducing morbidity and mortality and potentially improving survival. PMID- 12895397 TI - Dermatomyositis and Polymyositis. AB - Dermatomyositis (DM) and polymyositis (PM) are idiopathic inflammatory myopathies characterized by proximal greater than distal muscle weakness, elevated serum creatine kinase levels, electrophysiologic abnormalities, and inflammation on muscle biopsy. Clinically and electrophysiologically, DM and PM appear very similar, and muscle biopsy is the gold standard for diagnosis. Much of the PM literature based the diagnosis on Bohan and Peter's criteria, which is now obsolete given the advances of immunopathology. As diagnostic criteria for the inflammatory myopathies have been refined, it has become apparent that PM is much less common than previously thought, and, in fact, is probably quite rare. More recent literature, using strict histopathologic criteria for diagnosis of PM, has brought into question previously reported associations. Because of this, the clinical entity of PM is poorly defined. The exact incidence of each is unknown because previous epidemiologic studies often grouped them together, but overall the annual incidence of the inflammatory myopathies is approximately one in 100,000. DM and PM respond to immunomodulating therapies. High-dose oral prednisone is generally accepted first-line therapy. In patients who do not respond adequately to prednisone alone, or in whom prednisone cannot be weaned, methotrexate or azathioprine can be added. In the authors' experience, methotrexate works faster and is more effective than azathioprine. However, because of the increased risk of interstitial lung disease with methotrexate, the authors avoid this in patients with anti-Jo-1 antibodies and, obviously, in patients who already have pulmonary disease. If patients do not respond adequately to the combination of prednisone and methotrexate or azathioprine, a trial of intravenous immunoglobulin is administered. PMID- 12895396 TI - Monitoring bcr-abl by polymerase chain reaction in the treatment of chronic myeloid leukemia. AB - The elucidation of the molecular biology of chronic myeloid leukemia (CML) has provided a paradigm for understanding leukemogenesis, targeted drug development, and disease monitoring at the molecular level. Minimal residual disease (MRD) monitoring by fluorescence in situ hybridization and polymerase chain reaction (PCR) has become an important tool in predicting relapse after allogeneic transplant, allowing for early intervention strategies such as donor lymphocyte infusion. MRD monitoring is important for assessment of disease status in patients who obtain a complete cytogenetic remission, and this approach is likely to play an important role in following patients to determine who will relapse on imatinib mesylate therapy. This review focuses primarily on MRD monitoring by PCR. PMID- 12895398 TI - Chronic Inflammatory Demyelinating Polyneuropathy. AB - Although there are close to 10 randomized trials showing efficacy for prednisone, intravenous immunoglobulin, or plasmapheresis in chronic inflammatory demyelinating polyneuropathy (CIDP), large differences in cost, side effect profiles, and ease of use create controversy over the therapy that is best. Most clinicians use intravenous immunoglobulin or prednisone as first-line therapy. Unfortunately, the clinical trials performed to date are not easily extrapolated to answer, "which agent is best for clinical practice" because they have used varying doses and duration of therapy, different diagnostic criteria for CIDP, and mixtures of patients who have failed previous therapy versus those with newly diagnosed disease. There are a number of small case series describing the efficacy of immune-modulating agents, such as azathioprine, cyclophosphamide, cyclosporine A, interferon-alpha, and mycophenolate mofetil. These studies generally use these medications as second-line agents for patients who failed corticosteroids or intravenous immunoglobulin. In the authors' practice, this lack of robust knowledge still relegates these to second-line or prednisone sparing agents. PMID- 12895399 TI - Potential Role of Neuroprotective Agents in the Treatment of Patients with Acute Ischemic Stroke. AB - Currently, intravenous recombinant tissue plasminogen activator is the only US Food and Drug Administration-approved therapy for acute ischemic stroke. Although efficacious, its usefulness is limited, mainly because of the very limited time window for its administration. Neuroprotective treatments are therapies that block the cellular, biochemical, and metabolic elaboration of injury during or after exposure to ischemia, and have a potential role in ameliorating brain injury in patients with acute ischemic stroke. More than 50 neuroprotective agents have reached randomized human clinical trials in focal ischemic stroke, but none has been unequivocally proven efficacious, despite successful preceding animal studies. The failed neuroprotective trials of the past have greatly increased understanding of the fundamental biology of ischemic brain injury and have laid a strong foundation for future advance. Moreover, the recent favorable results of human clinical trials of hypothermia in human cardiac arrest and global brain ischemia have validated the general concept of neuroprotection for ischemic brain injury. Recent innovations in strategies of preclinical drug development and clinical trial design that rectify past defects hold great promise for neuroprotective investigation, including novel approaches to accelerating time to initiation of experimental treatment, use of outcome measures sensitive to treatment effects, and trial testing of combination therapies rather than single agents alone. Although no neuroprotective agent is of proven benefit for focal ischemic stroke, several currently available interventions have shown promising results in preliminary trials and may be considered for cautious, off-label use in acute stroke, including hypothermia, magnesium sulfate, citicoline, albumin, and erythropoietin. Overall, the prospects for safe and effective neuroprotective therapies to improve stroke outcome remain promising. PMID- 12895400 TI - Thrombolysis in the Treatment of Acute Ischemic Stroke. AB - Unfortunately, intravenous alteplase, when administered within 3 hours of symptom onset, remains the only US Food and Drug Administration-approved treatment for acute ischemic stroke; it is now 7 years since its approval. Intra-arterial thrombolysis of middle cerebral and basilar artery occlusions within 6 hours of stroke onset probably is efficacious also, but the clinical trial evidence supporting this claim is less robust. The whole field of acute stroke intervention is in urgent need of more randomized, controlled clinical trials to advance management. There are several promising avenues of investigation, but clinicians must remain cautious and skeptical until convincing proof of the added benefit of new interventions is obtained. PMID- 12895401 TI - Atherosclerotic Carotid Artery Occlusion. AB - Patients with acute stroke caused by atherosclerotic carotid artery occlusion (ACAO) should receive intravenous tissue plasminogen activator if they meet eligibility criteria. Patients with acute stroke caused by ACAO who are not eligible for intravenous tissue plasminogen activator should receive aspirin. Heparin or heparin-like drugs do not improve outcome and should not be used. Therapy for prevention of recurrent stroke in patients with ACAO should consist of lifestyle modifications, risk factor intervention, and aspirin. Other antiplatelet drugs should be considered in patients with contraindication to aspirin. Warfarin is not indicated. Extracranial-intracranial bypass surgery provides no benefit over medical therapy in preventing recurrent stroke in a general population of patients with ACAO or in any subgroups selected by clinical or arteriographic criteria. Extracranial-intracranial bypass surgery in patients selected by hemodynamic criteria should only be performed as part of a randomized controlled clinical trial. Other surgical or endovascular procedures have no proven value in treating or preventing stroke caused by ACAO. Asymptomatic carotid occlusion has a benign prognosis, and requires no specific treatment other than lifestyle modification and risk factor intervention. PMID- 12895402 TI - Cerebrovascular Disease in Pregnancy. AB - Stroke during pregnancy is a special category of stroke in young women. Although the absolute risk is small, there are diverse causes, including those inherent to the pregnant state, that may have a significant impact on maternal and fetal outcome. Severe pre-eclampsia and eclampsia are commonly associated with ischemic and hemorrhagic stroke, but must not be presumed the sole cause of stroke in pregnant women. Magnesium sulfate is the treatment of choice to prevent eclampsia. Randomized clinical trials in pregnant women are not available to provide guidance for the treatment of ischemic and hemorrhagic stroke in pregnant women. Various antithrombotic agents may be safely used during specific stages of pregnancy for treatment and prevention of ischemic stroke, with low-dose aspirin, unfractionated heparin, and low molecular weight heparin the preferred agents. Low molecular weight heparin may be safer than unfractionated heparin. Treatment of parenchymatous intracerebral hemorrhage and subarachnoid hemorrhage during pregnancy and the puerperium must be individualized. Aneurysms may be treated with neurosurgical clipping or endovascular coiling, depending on neurosurgical considerations. Cesarean or vaginal delivery may be used depending on the timing of delivery, adequacy of aneurysm occlusion, and risk to mother and fetus. Arteriovenous malformations are best treated in a multimodal fashion at a specialized treatment center. PMID- 12895403 TI - Cerebral Venous Thrombosis. AB - Because of its wide range of presentations, its highly variable mode of onset, its numerous causes, and its unpredictable outcome, cerebral venous thrombosis (CVT) remains a diagnostic and therapeutic challenge. Treatment of CVT consists primarily of symptomatic treatment of seizures and intracranial hypertension, antithrombotics, and etiologic treatment whenever possible. Heparin remains the first line of treatment for CVT; although its systematic use remains debated, recent studies have confirmed its safety even in patients with large hemorrhagic infarctions. The addition of local thrombolysis is indicated for patients with clinical worsening related to extension of the venous thrombosis, despite adequate anticoagulation and optimal symptomatic and etiologic treatment. In contrast to arterial stroke, complete recovery of prolonged or severe neurologic deficit is possible, justifying initiation of anticoagulation and eventually thrombolysis, even when the clinical situation seems desperate. New techniques using mechanical devices disrupting the clot may be used in addition to thrombolysis in rare cases. Ventricular drainage is indicated in cases of cerebellar infarction or deep venous thrombosis associated with hydrocephalus. Decompressive craniotomy may be performed acutely in patients with untractable intracranial hypertension and herniation. PMID- 12895404 TI - Relationship between donor animal age, follicular fluid steroid content and oocyte developmental competence in the pig. AB - The developmental competence of oocytes recovered from the ovaries of slaughtered prepubertal and adult pigs was evaluated after in vitro maturation, parthenogenetic activation and culture in vitro. In addition, the effect of prepubertal and adult follicular fluid (FF) on the developmental competence of prepubertal and adult oocytes was investigated. When matured in adult FF, the rates of cleavage (92 v. 73% P < 0.01) and blastocyst formation (57 v. 38%; P < 0.05) were greater for adult oocytes than for prepubertal oocytes. Blastocysts derived from adult oocytes had more trophectoderm cells (43 v. 30; P < 0.05) and total cells (51 v. 36; P < 0.05) than blastocysts derived from prepubertal oocytes. The developmental competence of prepubertal oocytes was not affected by the FF donor age, whereas the developmental competence of adult oocytes was. Blastocysts derived from adult oocytes matured in adult FF had more trophectoderm cells (38 v. 24; P < 0.005), inner cell mass cells (7 v. 3; P < 0.01) and total cells (45 v. 27; P < 0.001) than blastocysts derived from adult oocytes matured in prepubertal FF. Characterization of the steroid content of the FF used to supplement the maturation medium revealed that adult FF contained more progesterone (42 v. 23 ng mL(-1); P < 0.005) and androstenedione (70 v. 16 ng mL( 1); P < 0.05) than prepubertal FF. In addition, the molar ratios of progesterone to androstenedione, androstenedione to 17beta-oestradiol and androstenedione to testosterone differed (P < 0.05) between prepubertal and adult FF. The results support the hypothesis that a greater proportion of adult oocytes than of prepubertal oocytes has completed 'oocyte capacitation'. The differences in FF steroid content are indicative of the different follicular environments from which the prepubertal and adult oocytes were isolated, and may be attributed to the observed effects on oocyte developmental competence. PMID- 12895405 TI - Effect of ageing on the proliferation and apoptosis of testicular germ cells in the Syrian hamster Mesocricetus auratus. AB - The cellular mechanisms implicated in the atrophy of seminiferous epithelium in ageing are currently under debate, although recent reports suggest that apoptosis may be the primary mechanism implicated in aged germ cell loss. Other investigators have suggested that changes in spermatogonial proliferation are also involved. In the present work, the changes in proliferation and apoptosis in the seminiferous epithelium of aged (24 months) Syrian hamsters were examined in concert and compared with those in young (6 months) animals. Proliferation of germ cells was studied by bromodeoxyuridine labelling and apoptosis was assessed by transmission electron microscopy and in situ TUNEL labelling. Aged animals showed a significant decrease in the numbers of total and proliferating spermatogonia plus preleptotene spermatocytes per unit volume and per testis and in the proliferative index (24.8 +/- 1.6%) compared with young animals (30.8 +/- 1.2%) (P < 0.05). The number of apoptotic spermatogonia plus spermatocytes per unit volume and the apoptotic index were significantly higher in aged animals (1.51 +/- 0.23% v. 0.77 +/- 0.04%; P < 0.05). Apoptosis was confirmed by morphological characteristics: condensation of the chromatin and nuclear fragmentation. In aged hamsters, tubular degeneration could be classified into several categories, showing an increase of apoptotic cells in tubular cross sections characterized by maturation arrest in comparison with all other types. Spermatogonial proliferation was also diminished as seen in tubular cross sections showing hypospermatogenesis, sloughing off of germ cells and maturation arrest. The results obtained in the present study suggest that the decrease in the proliferation of spermatogonia and the increase in apoptosis constitute two consecutive mechanisms correlated with the ageing of the seminiferous epithelium. PMID- 12895406 TI - Expression of hyaluronan synthase-3 in porcine oviducal epithelium during oestrus. AB - Hyaluronan (HA) has been related to fertilization and embryo development in the pig. Furthermore, HA is present in pig oviduct fluid and the lining epithelium, particularly of the pre-ovulatory sperm reservoir. Because the mechanisms that regulate HA synthesis have not yet been clarified, semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) was conducted to assess the expression of mRNAs of two HA-synthesizing enzymes (has2 and has3) in the oviduct epithelium (uterotubal junction, isthmus, ampullary-isthmic junction and ampulla segments) of non-inseminated (control) and inseminated (treatment) sows at pre-, peri- and post-ovulatory oestrus. Only has3 mRNA was detected; it was present in all tubal segments of both control and treatment samples. The level of has3 expression did not vary significantly between non-inseminated and inseminated specimens, but there was a tendency (NS) for increased mean values during the peri- and post-ovulatory stages compared with pre-ovulation. It is concluded that has3 is expressed by the porcine endosalpinx epithelium and the levels of expression do not vary during the critical periods of sperm transport and fertilization, despite fluctuating levels of HA in the tubal fluid at corresponding periods. PMID- 12895407 TI - Expression of collagen XVIII mRNA and protein in human umbilical vein and placenta. AB - Endostatin is a potent angiogenic inhibitor that is derived from collagen XVIII by proteolytic cleavage. Localization of collagen XVIII has been reported in the basement membrane of blood vessels. To examine the involvement of collagen XVIII/endostatin during pregnancy, the distribution of collagen XVIII/endostatin protein in human umbilical vein was evaluated by immunohistochemistry. The expression of collagen XVIII/endostatin in cultured human umbilical vein endothelial cells (HUVEC) was also examined by immunocytochemistry and Northern blot analysis. To examine the release of endostatin in vivo and in vitro, concentrations of endostatin in umbilical venous blood and in HUVEC culture medium were determined using an enzyme-linked immunosorbent assay. Collagen XVIII/endostatin protein was localized to endothelial cells and their basement membrane in the umbilical vein. The expression of collagen XVIII mRNA and protein was detected in HUVEC. However, endostatin was not detected in umbilical venous blood or in HUVEC culture medium. The absence of endostatin release and the presence of its parental protein, collagen XVIII, suggest that the cleavage mechanisms of endostatin might be strongly inhibited under the physiological conditions present during pregnancy. It is therefore considered that vasculature in the feto-placental unit is highly angiogenic, even at the time of parturition. PMID- 12895408 TI - Oocyte competence in repeat-breeder heifers: effects of an optimized ovum pick-up schedule on expression of oestrus, follicular development and fertility. AB - Repeat-breeder heifers (RBH) have been shown to present reproductive perturbations during spontaneous cyclicity, which affects oestrus and ovulation. Some of these disturbances (e.g. deviating hormone patterns) are also present during and after cycles of twice-weekly ovum pick-up (o.p.u.), performed according to an optimized schedule allowing normal oestrous cyclicity. In the present study, the effects of o.p.u. on oocyte competence in in vitro maturation (i.v.m.) and in vitro fertilization (i.v.f.) have been evaluated, as were the effects on expression of oestrus and fertility in five RBH (> or =4 artificial inseminations) and five virgin heifers (VH controls). In total, 269 RBH and 174 VH oocytes were scored for quality prior to i.v.m. and i.v.f. The number of follicles available for puncture was higher in RBH, but the oocyte recovery rate after o.p.u. was lower in RBH compared with VH controls and the recovered RBH oocytes were of lower quality, as judged by their appearance at retrieval. Confocal laser scanning and transmission electron microscopy of immature oocytes did not reveal any differences between RBH and VH control oocytes with respect to nuclear and mitochondrial status. However, after i.v.m., the cytoplasmic spatial reorganization of mitochondria and cortical granules was less advanced in RBH, which could contribute to the subfertility that defines the syndrome. Cleavage rates after i.v.f. were similar in RBH and VH controls. Subsequent to the o.p.u. period, in vivo fertility after controlled artificial insemination was comparable with field fertility rates in both RBH and VH. PMID- 12895409 TI - Fetal mortality diagnosis by ultrasound in the vicuna (Vicugna vicugna). AB - Ultrasonography is widely used in domestic species of camelids, but there is no information about the use of this technique for pregnancy diagnosis and determination of embryonic or fetal losses in the vicuna (Vicugna vicugna). The study was performed in 202 vicunas (3-year-old females, n = 31; adult females, n = 171) mated during the summer months (January through March 2001) at the Abra Pampa Experimental Farm of Altitude in north-west Argentina. Transrectal ultrasound examination was performed in May (estimated 40-120 days of gestation) to determine the number of pregnant females. The pregnancy rate was 45.5% (92/202). No significant difference (P > 0.05) was observed between the pregnancy rate of 3-year-old females (41.9%) and adult females (46.2%). In December (estimated 250-330 days of gestation) of the same year, a second ultrasonographic study was performed on those vicunas that were diagnosed as pregnant from the first ultrasound scan. Of 92 animals diagnosed as pregnant in May, only 84 were present in December, because eight females died in the period of study. Overall, 11.9% (10/84) of fetuses were lost during the period (18.1% in 3-year-old vicunas and 10.9% in adult vicunas). In conclusion, transrectal ultrasonography was found to provide a rapid and non-invasive means for pregnancy and fetal mortality diagnosis in vicunas. PMID- 12895410 TI - Ubiquitin-specific protease activity of USP9Y, a male infertility gene on the Y chromosome. AB - Deletions of USP9Y have been observed among infertile males with defective spermatogenesis. Therefore, the gene has been designated as a male infertility gene on the Y chromosome. However, it remains to be determined how male infertility results from deletions of this gene. In order to initiate an investigation into the cellular functions of USP9Y in male germ cell development, in the present study we characterized the enzymatic specificity of USP9Y. Our results show that both USP9Y and Fam, the mouse infertility protein Usp9x, possess a protease activity specific to ubiquitin. These results suggest that, through de-ubiquitination, USP9Y may stabilize a specific target protein that is important for male germ cell development. PMID- 12895411 TI - Parthenogenetic activation of rat oocytes and their development (in vitro). AB - The present study was performed to determine suitable methods for parthenogenetic activation and subsequent development of rat oocytes in vitro. In the first series of experiments, the ability of electrical pulses, strontium, ethanol and ionomycin to activate Sprague-Dawley (SD) rat oocytes was examined. The synergistic effect of strontium and cycloheximide or puromycin was also examined in the second series of experiments. In the third series of experiments, the development of F1 hybrid (SD x Dark Agouti) parthenotes activated with different concentrations of strontium (10-0.08 mM) was compared with that of SD parthenotes. The effect of the timing of activation (10 min and 2, 4 and 6 h after cervical dislocation) was also assessed in a fourth series of experiments. The oocytes activated by strontium showed higher pronuclear formation and cleavage rates than those in the other groups (P < 0.05). Higher blastocyst development was obtained from parthenotes activated by strontium and strontium cycloheximide compared with the strontium-puromycin group (P < 0.01). However, the total cell number of blastocysts from the strontium-cycloheximide activation group was higher than that of other groups (P < 0.05). With strontium (2.5-10 mM) treatment, 40.9% of blastocysts were obtained from F1 hybrid oocytes, whereas 22.9% were obtained from SD (P < 0.01). The oocytes activated 10 min or 2 h following cervical dislocation showed higher blastocyst development than those of the 4 and 6 h groups (P < 0.01). These results suggest that strontium cycloheximide produces the highest parthenogenetic activation rate in the rat and that oocytes must be activated by 2 h after cervical dislocation. PMID- 12895412 TI - Stress hormone enhances synaptic NMDA response on dopamine neurons. PMID- 12895413 TI - Thalamocortical topography reloaded: it's not where you go, but how you get there. PMID- 12895414 TI - "Resistant" channels reluctantly reveal their roles. PMID- 12895415 TI - Answers and questions from the KvAP structures. AB - The recent landmark structures of KvAP, a voltage-gated potassium (Kv) channel, provide the first high-resolution experimental structural models of this class of proteins. Previous extensive studies of Kv channels provide a means to evaluate and interpret the KvAP structures. In this minireview, we survey different experimental approaches to Kv channels and map these findings to KvAP, showing that the relationship between the KvAP structures and other Kv channels is uncertain. PMID- 12895416 TI - Corticotropin-releasing factor requires CRF binding protein to potentiate NMDA receptors via CRF receptor 2 in dopamine neurons. AB - Stress increases addictive behaviors and is a common cause of relapse. Corticotropin-releasing factor (CRF) plays a key role in the modulation of drug taking by stress. However, the mechanism by which CRF modulates neuronal activity in circuits involved in drug addiction is poorly understood. Here we show that CRF induces a potentiation of NMDAR (N-methyl-D-aspartate receptor)-mediated synaptic transmission in dopamine neurons of the ventral tegmental area (VTA). This effect involves CRF receptor 2 (CRF-R2) and activation of the phospholipase C (PLC)-protein kinase C (PKC) pathway. We also find that this potentiation requires CRF binding protein (CRF-BP). Accordingly, CRF-like peptides, which do not bind the CRF-BP with high affinity, do not potentiate NMDARs. These results provide evidence of the first specific roles for CRF-R2 and CRF-BP in the modulation of neuronal activity and suggest that NMDARs in the VTA may be a target for both drugs of abuse and stress. PMID- 12895417 TI - Triple-transgenic model of Alzheimer's disease with plaques and tangles: intracellular Abeta and synaptic dysfunction. AB - The neuropathological correlates of Alzheimer's disease (AD) include amyloid-beta (Abeta) plaques and neurofibrillary tangles. To study the interaction between Abeta and tau and their effect on synaptic function, we derived a triple transgenic model (3xTg-AD) harboring PS1(M146V), APP(Swe), and tau(P301L) transgenes. Rather than crossing independent lines, we microinjected two transgenes into single-cell embryos from homozygous PS1(M146V) knockin mice, generating mice with the same genetic background. 3xTg-AD mice progressively develop plaques and tangles. Synaptic dysfunction, including LTP deficits, manifests in an age-related manner, but before plaque and tangle pathology. Deficits in long-term synaptic plasticity correlate with the accumulation of intraneuronal Abeta. These studies suggest a novel pathogenic role for intraneuronal Abeta with regards to synaptic plasticity. The recapitulation of salient features of AD in these mice clarifies the relationships between Abeta, synaptic dysfunction, and tangles and provides a valuable model for evaluating potential AD therapeutics as the impact on both lesions can be assessed. PMID- 12895418 TI - Local tissue interactions across the dorsal midline of the forebrain establish CNS laterality. AB - The mechanisms that establish behavioral, cognitive, and neuroanatomical asymmetries are poorly understood. In this study, we analyze the events that regulate development of asymmetric nuclei in the dorsal forebrain. The unilateral parapineal organ has a bilateral origin, and some parapineal precursors migrate across the midline to form this left-sided nucleus. The parapineal subsequently innervates the left habenula, which derives from ventral epithalamic cells adjacent to the parapineal precursors. Ablation of cells in the left ventral epithalamus can reverse laterality in wild-type embryos and impose the direction of CNS asymmetry in embryos in which laterality is usually randomized. Unilateral modulation of Nodal activity by Lefty1 can also impose the direction of CNS laterality in embryos with bilateral expression of Nodal pathway genes. From these data, we propose that laterality is determined by a competitive interaction between the left and right epithalamus and that Nodal signaling biases the outcome of this competition. PMID- 12895419 TI - Neurogenin2 specifies the connectivity of thalamic neurons by controlling axon responsiveness to intermediate target cues. AB - Many lines of evidence indicate that important traits of neuronal phenotype, such as cell body position and neurotransmitter expression, are specified through complex interactions between extrinsic and intrinsic genetic determinants. However, the molecular mechanisms specifying neuronal connectivity are less well understood at the transcriptional level. Here we demonstrate that the bHLH transcription factor Neurogenin2 cell autonomously specifies the projection of thalamic neurons to frontal cortical areas. Unexpectedly, Ngn2 determines the projection of thalamic neurons to specific cortical domains by specifying the responsiveness of their axons to cues encountered in an intermediate target, the ventral telencephalon. Our results thus demonstrate that in parallel to their well-documented proneural function, bHLH transcription factors also contribute to the specification of neuronal connectivity in the mammalian brain. PMID- 12895420 TI - Area specificity and topography of thalamocortical projections are controlled by ephrin/Eph genes. AB - The mechanisms generating precise connections between specific thalamic nuclei and cortical areas remain poorly understood. Using axon tracing analysis of ephrin/Eph mutant mice, we provide in vivo evidence that Eph receptors in the thalamus and ephrins in the cortex control intra-areal topographic mapping of thalamocortical (TC) axons. In addition, we show that the same ephrin/Eph genes unexpectedly control the inter-areal specificity of TC projections through the early topographic sorting of TC axons in an intermediate target, the ventral telencephalon. Our results constitute the first identification of guidance cues involved in inter-areal specificity of TC projections and demonstrate that the same set of mapping labels is used differentially for the generation of topographic specificity of TC projections between and within individual cortical areas. PMID- 12895421 TI - Atomic proximity between S4 segment and pore domain in Shaker potassium channels. AB - A recently proposed model for voltage-dependent activation in K+ channels, largely influenced by the KvAP X-ray structure, suggests that S4 is located at the periphery of the channel and moves through the lipid bilayer upon depolarization. To investigate the physical distance between S4 and the pore domain in functional channels in a native membrane environment, we engineered pairs of cysteines, one each in S4 and the pore of Shaker channels, and identified two instances of spontaneous intersubunit disulfide bond formation, between R362C/A419C and R362C/F416C. After reduction, these cysteine pairs bound Cd2+ with high affinity, verifying that the residues are in atomic proximity. Molecular modeling based on the MthK structure revealed a single position for S4 that was consistent with our results and many other experimental constraints. The model predicts that S4 is located in the groove between pore domains from different subunits, rather than at the periphery of the protein. PMID- 12895422 TI - Functional specialization of presynaptic Cav2.3 Ca2+ channels. AB - Ca2+ influx into presynaptic terminals via voltage-dependent Ca2+ channels triggers fast neurotransmitter release as well as different forms of synaptic plasticity. Using electrophysiological and genetic techniques we demonstrate that presynaptic Ca2+ entry through Cav2.3 subunits contributes to the induction of mossy fiber LTP and posttetanic potentiation by brief trains of presynaptic action potentials while they do not play a role in fast synaptic transmission, paired-pulse facilitation, or frequency facilitation. This functional specialization is most likely achieved by a localization remote from the release machinery and by a Cav2.3 channel-dependent facilitation of presynaptic Ca2+ influx. Thus, the presence of Cav2.3 channels boosts the accumulation of presynaptic Ca2+ triggering presynaptic LTP and posttetanic potentiation without affecting the low release probability that is a prerequisite for the enormous plasticity displayed by mossy fiber synapses. PMID- 12895423 TI - Hypotonicity induces TRPV4-mediated nociception in rat. AB - We hypothesized that TRPV4, a member of the transient receptor family of ion channels, functions as a sensory transducer for osmotic stimulus-induced nociception. We found that, as expected for a transducer molecule, TRPV4 protein is transported in sensory nerve distally toward the peripheral nerve endings. In vivo single-fiber recordings in rat showed that hypotonic solution activated 54% of C-fibers, an effect enhanced by the hyperalgesic inflammatory mediator prostaglandin E2. This osmotransduction causes nociception, since administration of a small osmotic stimulus into skin sensitized by PGE2 produced pain-related behavior. Antisense-induced decrease in expression of TRPV4 confirmed that the channel is required for hypotonic stimulus-induced nociception. Thus, we conclude that TRPV4 can function as an osmo-transducer in primary afferent nociceptive nerve fibers. Because this action is enhanced by an inflammatory mediator, TRPV4 may be important in pathological states and may be an attractive pharmacological target for the development of novel analgesics. PMID- 12895424 TI - Narp and NP1 form heterocomplexes that function in developmental and activity dependent synaptic plasticity. AB - Narp is a neuronal immediate early gene that plays a role in excitatory synaptogenesis. Here, we report that native Narp in brain is part of a pentraxin complex that includes NP1. These proteins are covalently linked by disulfide bonds into highly organized complexes, and their relative ratio in the complex is dynamically dependent upon the neuron's activity history and developmental stage. Complex formation is dependent on their distinct N-terminal coiled-coil domains, while their closely homologous C-terminal pentraxin domains mediate association with AMPA-type glutamate receptors. Narp is substantially more effective in assays of cell surface cluster formation, coclustering of AMPA receptors, and excitatory synaptogenesis, yet their combined expression results in supraadditive effects. These studies support a model in which Narp can regulate the latent synaptogenic activity of NP1 by forming mixed pentraxin assemblies. This mechanism appears to contribute to both activity-independent and activity dependent excitatory synaptogenesis. PMID- 12895425 TI - Synaptic vesicle pools at the frog neuromuscular junction. AB - We have characterized the morphological and functional properties of the readily releasable pool (RRP) and the reserve pool of synaptic vesicles in frog motor nerve terminals using fluorescence microscopy, electron microscopy, and electrophysiology. At rest, about 20% of vesicles reside in the RRP, which is depleted in about 10 s by high-frequency nerve stimulation (30 Hz); the RRP refills in about 1 min, and surprisingly, refilling occurs almost entirely by recycling, not mobilization from the reserve pool. The reserve pool is depleted during 30 Hz stimulation with a time constant of about 40 s, and it refills slowly (half-time about 8 min) as nascent vesicles bud from randomly distributed cisternae and surface membrane infoldings and enter vesicle clusters spaced at regular intervals along the terminal. Transmitter output during low-frequency stimulation (2-5 Hz) is maintained entirely by RRP recycling; few if any vesicles are mobilized from the reserve pool. PMID- 12895426 TI - Noncanonical signaling by ionotropic kainate receptors. AB - The potent neurotoxin kainate activates ion channel-forming receptors. However, it can also activate a G protein-coupled signaling pathway to inhibit transmitter release in central neurons. It remains unclear whether the same receptor complex is involved in both signaling activities. Here we show that in a population of dorsal root ganglion cells, exposure to kainate elicits a G protein-dependent increase in intracellular Ca2+. Furthermore, in these cells a brief exposure to kainate inhibited the K+-induced Ca2+ increase, a process that was sensitive to the G protein inhibitor Pertussis toxin and inhibitors of protein kinase C. This metabotropic action did not require ion channel activity and was not observed in neurons prepared from mice deficient for the ion channel-forming subunit GluR5. These results indicate that GluR5, an ion channel-forming subunit, signals through a second messenger cascade, inhibiting voltage-dependent Ca2+ channels. Thus, such a system represents a noncanonical signaling route of ion channel forming receptors. PMID- 12895427 TI - Stereopsis activates V3A and caudal intraparietal areas in macaques and humans. AB - Stereopsis, the perception of depth from small differences between the images in the two eyes, provides a rich model for investigating the cortical construction of surfaces and space. Although disparity-tuned cells have been found in a large number of areas in macaque visual cortex, stereoscopic processing in these areas has never been systematically compared using the same stimuli and analysis methods. In order to examine the global architecture of stereoscopic processing in primate visual cortex, we studied fMRI activity in alert, fixating human and macaque subjects. In macaques, we found strongest activation to near/far compared to zero disparity in areas V3, V3A, and CIPS. In humans, we found strongest activation to the same stimuli in areas V3A, V7, the V4d topolog (V4d-topo), and a caudal parietal disparity region (CPDR). Thus, in both primate species a small cluster of areas at the parieto-occipital junction appears to be specialized for stereopsis. PMID- 12895428 TI - Interaction of retinal image and eye velocity in motion perception. AB - When we move our eyes, why does the world look stable even as its image flows across our retinas, and why do afterimages, which are stationary on the retinas, appear to move? Current theories say this is because we perceive motion by summation: if an object slips across the retina at r degrees/s while the eye turns at e degrees/s, the object's perceived velocity in space should be r + e. We show that activity in MT+, the visual-motion complex in human cortex, does reflect a mix of r and e rather than r alone. But we show also that, for optimal perception, r and e should not summate; rather, the signals coding e interact multiplicatively with the spatial gradient of illumination. PMID- 12895429 TI - Enhanced vasoconstrictor responses in eNOS deficient mice. AB - Previous studies suggest that vasoconstriction is modulated by nitric oxide (NO). Contractions to ET-1 and/or thromboxane may be enhanced during chronic deficiency in expression or activity of NO synthase (NOS). Multiple isoforms of NOS are expressed within the vessel wall and purely pharmacological approaches cannot define the role of each. We tested the hypothesis that vasoconstriction to endothelin-1 (ET-1) and/or the thromboxane mimetic, U46619, is enhanced under conditions of chronic, selective deficiency in endothelial NOS (eNOS-/-) by examining responses in aorta from eNOS-/- mice compared to wild type (eNOS+/+). ET-1 produced dose-dependent contraction of aorta from eNOS+/+ mice that was increased twofold following acute inhibition of all NOS isoforms with N(G)-nitro L-arginine (L-NNA). In eNOS-/- mice, contractions to ET-1 were increased twofold compared to eNOS+/+. L-NNA had no effect. Although contraction of the aorta to thromboxane mimetic U46619 was increased at lower concentrations, maximal contractions to U46619 were not increased following acute inhibition of NOS or in eNOS-/- mice. These studies provide direct evidence that vasoconstriction to ET-1 and thromboxane is augmented in the face of eNOS deficiency, demonstrating that eNOS normally inhibits vascular contractile responses. PMID- 12895430 TI - Peroxynitrite and NO+ donors form colored nitrite adducts with sinapinic acid: potential applications. AB - Sinapinic acid (3,5-dimethoxy-4-hydroxycinnamic acid, SA) reacted with peroxynitrous acid at neutral pH with a second-order rate constant of 812 M(-1)s( 1), to yield a red product (lambda(max), 532 nm). The identical colored product could be formed with acidified decomposed peroxynitrous acid solutions or nitrite at slower rates (0.1M HCl, 8.32 M(-1)s(-1); 10% acetic acid, 0.0004 M(-1)s(-1)). The red compound is thought to be O-nitrososinapinic acid (3,5-dimethoxy-4 nitrosooxycinnamic acid) which can be formed by reaction with either peroxynitrous acid or nitrous acid. The extinction coefficient of O nitrososinapinic acid (ONSA) was estimated to be 8419 M(-1)cm(-1) at 510 nm in 10% acetic acid and 90% acetonitrile. ONSA was also formed via NO(+) transfer from S-nitrosoglutathione (GSNO). ONSA in turn can S-nitrosate low molecular weight thiols and protein thiols. SA was also shown to act as a peroxynitrite sink as it effectively prevented the oxidation of dihydrorhodamine under physiological conditions. The fact that O-nitrososinapinic acid is stable and can be used to S-nitrosate thiol containing amino acids, peptides, and proteins makes it a potentially useful reagent in the study of S-nitrosothiol biochemistry and physiology. In addition, the relatively high extinction coefficient of O nitrososinapinic acid means that it could be utilized as an analyte for the spectroscopic detection of peroxynitrite or NO(+)-donors in the submicromolar range. PMID- 12895431 TI - Cell density-enhanced expression of inducible nitric oxide synthase in murine macrophages mediated by interferon-beta. AB - Nitric oxide (NO) has an important cytotoxic role in host defense processes against invading microorganisms and neoplastic cells. Here we demonstrate the effect of culture density on the expression of NO synthase and NO production by lipopolysaccharide (LPS)-activated RAW 264.7 macrophages. At high cell densities, the LPS-induced expression of iNOS message, protein, and activity is markedly enhanced. We demonstrate the effects to be mediated by a diffusible macrophage product. Increasing cell density correlates with activation of IFN-dependent signaling pathways. We observe enhanced phosphorylation of STAT-1 on tyrosine 701 and serine 727, and an increase in STAT-1 DNA binding. Expression of the IFN stimulated transcription factor IRF-1 is also enhanced. The data are consistent with the reported involvement of IFN-beta as an autocrine co-activator of iNOS expression. Considering the importance of NO as a cytotoxic mediator of host immunity, the data suggest that macrophage density is important in regulating the magnitude of NO production, and thus, the host response to infection. PMID- 12895432 TI - Suppression of nitric oxide oxidation to nitrite by curcumin is due to the sequestration of the reaction intermediate nitrogen dioxide, not nitric oxide. AB - Curcumin, a phytochemical with antioxidant and other cytoprotective properties, has been reported to reduce nitrite formation during nitric oxide (NO) oxidation in solution. This decrease in nitrite production was attributed to the direct sequestration of NO by curcumin. In this report, we confirm that curcumin inhibits nitrite formation from DEA/NO-derived NO in a concentration-dependent manner. However, curcumin over a concentration range of 3-50 microM had no effect on the concentration of free NO (0.5 microM) in solution at 37 degrees C as assessed using an NO electrode. We conclude that the inhibitory effect of curcumin on the oxidation of NO to nitrite is due to its known sequestration of the reaction intermediate nitrogen dioxide (NO(2)). The ability of curcumin to sequester NO(2), but not NO, suggests that curcumin may be useful for separating the actions of NO(2) from those of NO in various biological systems. PMID- 12895433 TI - Gypenosides derived from Gynostemma pentaphyllum suppress NO synthesis in murine macrophages by inhibiting iNOS enzymatic activity and attenuating NF-kappaB mediated iNOS protein expression. AB - Gypenosides isolated from Gynostemma pentaphyllum are widely used in traditional Chinese medicine, with beneficial effects reported in numerous diseases, including inflammation and atherosclerosis, although the mechanism underlying these therapeutic effects is unknown. Because increased nitric oxide (NO) plays a role in these pathological conditions, we investigated whether the pharmacological activity of gypenosides is due to suppression of NO synthesis. The markedly increased production of nitrite by stimulation of RAW 264.7 murine macrophages with 1 microg/mL lipopolysaccharide (LPS) for 20 h (unstimulated: 0.3+/-0.3 microM vs. LPS: 32.5+/-1.2 microM) was dose-dependently inhibited by gypenosides (0.1-100 microg/mL). When cells were pretreated with gypenosides (for 1h) prior to LPS stimulation, subsequent NO production was significantly attenuated (IC(50) of 3.1+/-0.4 microg/mL) (P<0.05). Gypenosides (25 microg/mL) produced the same maximum inhibition of LPS-induced NO production as aminoguanidine, a standard inhibitor of NOS enzymes. Suppression of NO production occurred both by direct inhibition of the activity and expression of iNOS. Inhibition of iNOS protein expression appears to be at the transcriptional level, since gypenosides decreased LPS-induced NF-kappaB activity in a dose-dependent manner (P<0.05), with significant inhibition achieved following pretreatment with 10 microg/mL gypenoside. Taken together, these results suggest that gypenosides derived from G. pentaphyllum suppress NO synthesis in murine macrophages by inhibiting iNOS enzymatic activity and attenuating NF-kappaB-mediated iNOS protein expression, thereby implicating a mechanism by which gypenosides may exert their therapeutic effects. PMID- 12895434 TI - Electrochemical activation of electrodes for amperometric detection of nitric oxide. AB - An open question in the literature of nitric oxide detection was investigated: does electrochemical activation account for the enhanced properties of certain presumed chemically-modified electrodes? Uniform electrodes of graphite, iridium, palladium, platinum, and ruthenium were exposed to potential cycling and then tested for amperometric response to nitric oxide to identify principles that govern electrochemical activation of nitric oxide electrodes. These electrodes were compared to similar electrodes that were not cycled. Only cycled graphite and ruthenium showed significantly increased responses. Graphite demonstrated enhanced performance after exposure to cycling potentials at which oxygen, CO(2), and soluble carbonates form, suggesting that erosion of the electrode enhanced its response by increasing the surface area accessible to nitric oxide. This may explain the performance of carbon fibers cycled to the same potentials in solutions containing metalloporphyrins. The response of ruthenium was enhanced after cycling to less extreme potentials at which soluble species do not form and at which a metallic conductive oxide, RuO(2), could lay down a stable, adherent layer on the electrode surface. Cycled ruthenium also exhibited a much greater increase in capacitance after cycling, consistent with the formation of a conductive surface layer. PMID- 12895435 TI - Insulin enhances the expression of the endothelial nitric oxide synthase in native endothelial cells: a dual role for Akt and AP-1. AB - Insulin-induced vasodilatation in vivo has been attributed to the activation of the endothelial nitric oxide (NO) synthase (eNOS). The present study addressed the effects of insulin on the activity and expression of eNOS in native and cultured endothelial cells. Insulin applied to native porcine aortic endothelial cells elicited the tyrosine phosphorylation of the insulin receptor and receptor substrate, the subsequent activation of phosphatidylinositol 3-kinase (PI 3-K), Akt (protein kinase B), and ERK1/2. Insulin did not activate eNOS in cultured endothelial cells nor relax endothelium-intact arterial segments. However, 4h after application of insulin to native endothelial cells eNOS mRNA was increased 2-fold. A comparable increase in eNOS protein was detected after 18-24h and associated with an increase in intracellular cyclic GMP. In native endothelial cells, insulin enhanced the DNA-binding activity of Sp1 and AP-1, but not that of NF-kappaB. The insulin-induced increase in eNOS expression was prevented by wortmannin as well as by AP-1 decoy oligonucleotides. The MEK1 inhibitor, PD 98059, also enhanced eNOS expression in native and cultured endothelial cells, an effect which was independent of ERK1/2 and associated with an increase in the DNA binding activity of AP-1 and Sp1. These results demonstrate that insulin activates multiple signalling pathways in endothelial cells but does not acutely activate eNOS. Insulin however enhances eNOS mRNA and protein by a mechanism involving the combined activation of a PI 3-K- and AP-1-dependent pathway. PMID- 12895436 TI - Cyclic AMP regulate adenovirus mediated inducible nitric oxide synthase expression and CMV promoter activity in cultured hepatocytes. AB - We examined effects of cyclic AMP on adenovirus mediated inducible nitric oxide synthase expression in primary rat hepatocytes. In AdCMV-hiNOS infected hepatocytes, dbcAMP at a concentration of 1-100 microM inhibited nitrite and nitrate accumulation, iNOS protein and iNOS mRNA expression. At a concentration of 1-5mM, dbcAMP increased iNOS activity and expression. Both the inhibitory and stimulatory effects of cAMP on adenovirus mediated iNOS expression were dose dependent. This dual effect of cAMP on iNOS activity and expression can be accounted by its effect on CMV promoter activity since dbcAMP significantly inhibited CMV promoter activity in hepatocytes at low concentration and induced it at high concentration. We concluded that dbcAMP can regulate adenovirus mediated human iNOS expression in primary hepatocytes by the mechanisms regulating the CMV promoter used in the adenovirus vector. PMID- 12895437 TI - Repair of chronic spinal cord injury. AB - Advances in medical and rehabilitative care now allow the 10-12,000 individuals who suffer spinal cord injuries each year in the United States to lead productive lives of nearly normal life expectancy, so that the numbers of those with chronic injuries will approximate 300,000 at the end of the next decade. This signals an urgent need for new treatments that will improve repair and recovery after longstanding injuries. In the present report we consider the characteristics of the chronically injured spinal cord that make it an even more challenging setting in which to elicit regeneration than the acutely injured spinal cord and review the treatments that have been designed to enhance axon growth. When applied in the first 2 weeks after experimental spinal cord injury, transplants, usually in combination with supplementary neurotrophic factors, and possibly modifications of the inhibitory central nervous system environment, have produced limited long distance axon regeneration and behavioral recovery. When applied to injuries older than 4 weeks, the same treatments have almost invariably failed to overcome the obstacles posed by the neurons' diminished capacity for regeneration and by the increasing hostility to growth of the terrain at and beyond the injury site. Novel treatments that have stimulated regeneration after acute injuries have not yet been applied to chronic injuries. A therapeutic strategy that combines rehabilitation training and pharmacological modulation of neurotransmitters appears to be a particularly promising approach to increasing recovery after longstanding injury. Identifying patients with no hope of useful recovery in the early days after injury will allow these treatments to be administered as early as possible. PMID- 12895438 TI - Evaluation of MCAO stroke models in normotensive rats: standardized neocortical infarction by the 3VO technique. AB - The temporary three-vessel occlusion (3VO) technique with a surgical approach for middle cerebral artery (MCA) produces consistent cerebral infarction in the neocortex in normotensive rats. The intraluminal thread-occlusion technique with an endovascular approach targeting the MCA occlusion (MCAO) is more widely used since it does not require complicated intracranial procedures. The aim of this study was to review the methods/models for MCAO stroke in normotensive rats and to evaluate a 3VO stroke model that provides consistent degrees and variance of cortical stroke injury for additional discussion. First, we analyzed a model with modified temporary 3VO technique requiring less complicated procedures than the temporary 3VO model, i.e., temporary occlusion of the bilateral common carotid arteries (CCAs) superimposed on a permanent occlusion of the MCA, in Sprague Dawley rats or C57BL/6J mice. In the microvascular tissue (cerebral) perfusion study, significant reductions in regional cerebral perfusion during the 3VO accompanied a rapid return to baseline after release of the CCAs, showing that the technique induces temporary focal ischemia. The average sizes and variances of the neocortical infarction in this model, together with those in the other normotensive rat models caused by the 3VO technique in the literature, indicated a standard size and variance of infarcted lesion in the control groups relative to the specific ischemic period. However, stroke injuries in the neocortex induced by the thread occlusion technique showed greater variability with less consistent lesion sizes. Inclusion/exclusion criteria to avoid inappropriate cases with too mild (no/faint infarction) or too great (huge/fatal infarction) severity in the ischemic injury may differ between laboratories in the thread occlusion model. PMID- 12895439 TI - Hematogenous macrophages express CD8 and distribute to regions of lesion cavitation after spinal cord injury. AB - Historically, CD4 and CD8 antigens have been used to designate functionally distinct T-lymphocyte subsets. However, these antigens also have been described on macrophages in the normal and pathologic central nervous system (CNS). Signaling through CD4 or CD8 may impart unique functions in macrophage subsets that express these antigens. In the current study, the distribution and temporal patterns of expression of CD4 and CD8 were evaluated on various cell types within the traumatically injured spinal cord. The data reveal divergent patterns of CD4 and CD8 expression on unique macrophage populations. Specifically, we show sustained elevations of CD4 expression on microglia and macrophages throughout the lesion site and spared white matter. In contrast, CD8 is predominantly associated with hematogenous macrophages that are recruited from the blood during the first week postinjury. The distribution of CD8-positive cells is restricted to areas of necrotic cavitation. Differential signaling of resident and recruited macrophages through CD4 or CD8 may explain the apparent dichotomy of CNS macrophage-mediated injury and repair. PMID- 12895440 TI - Transplantation of porcine umbilical cord matrix cells into the rat brain. AB - Immune rejection of transplanted material is a potential complication of organ donation. In response to tissue transplantation, immune rejection has two components: a host defense directed against the grafted tissue and an immune response from the grafted tissue against the host (graft vs host disease). To treat immune rejection, transplant recipients are typically put on immunosuppression therapy. Complications may arise from immune suppression or from secondary effects of immunosuppression drugs. Our preliminary work indicated that stem cells may be xenotransplanted without immunosuppression therapy. Here, we investigated the survival of pig stem cells derived from umbilical cord mucous connective tissue (UCM) after transplantation into rats. Our data demonstrate that UCM cells survive at least 6 weeks without immune suppression of the host animals after transplantation into either the brain or the periphery. In the first experiment, UCM cells were transplanted into the rat brain and recovered in that tissue 2-6 weeks posttransplantation. At 4 weeks posttransplantation, the UCM cells engrafted into the brain along the injection tract. The cells were small and roughly spherical. The transplanted cells were positively immunostained using a pig-specific antibody for neuronal filament 70 (NF70). In contrast, 6 weeks posttransplantation, about 10% of the UCM cells that were recovered had migrated away from the injection site into the region just ventral to the corpus callosum; these cells also stained positively for NF70. In our second experiment, UCM cells that were engineered to constitutively express enhanced green fluorescent protein (eGFP) were transplanted. These cells were recovered 2-4 weeks after brain transplantation. Engrafted cells expressing eGFP and positively staining for NF70 were recovered. This finding indicates a potential for gene therapy. In the third experiment, to determine whether depositing the graft into the brain protected UCM cells from immune detection/clearance, UCM cells were injected into the tail vein and/or the semitendinosis muscle in a group of animals. UCM cells were recovered from the muscle or within the kidney 3 weeks posttransplantation. In control experiments, rat brains were injected with PKH 26 labeled UCM cells that had been lysed by repeated sonic disruption. One and 2 weeks following injection, no PKH 26-labeled neurons or glia were observed. Taken together, these data indicate that UCM cells can survive xenotransplantation and that a subset of the UCM cells respond to local signals to differentiate along a neural lineage. PMID- 12895441 TI - Novel intrathecal delivery system for treatment of spinal cord injury. AB - A novel, localized method for potential delivery of therapeutic agents to the injured spinal cord was investigated. The strategy consists of a polymeric drug solution that gels after injection into the subarachnoid space (SAS). By dispersing therapeutic agents in the polymeric solution, a method is provided for localized delivery to the spinal cord. To determine whether intrathecal injection of this drug delivery system (DDS) would affect cerebrospinal fluid (CSF) flow, a spinal canal model was built using dimensional analysis. Blocking up to 52% of the modeled subarachnoid space of the spinal canal caused minimal pressure differences (9.22 +/- 1.45 Pa), suggesting that implantation of a DDS would not subject the spinal cord to increased pressure. The safety of the DDS was also assessed in vivo by injecting collagen into the SAS of Sprague Dawley rats. Controls received injections of artificial CSF (aCSF). Collagen or aCSF was injected at the T2-T3 spinal level of both uninjured rats and rats injured with a 20g compression clip. The injected collagen persisted in the SAS for at least 8 weeks post-implantation and did not elicit an inflammatory reaction in either uninjured or injured animals. Long-term functional behavior was evaluated with the Basso, Beattie, and Bresnahan (BBB) scale weekly for 8 weeks. Functional behavior was similar in the collagen and aCSF groups, also indicating that the DDS was safe. This minimally invasive DDS may provide an alternative, safe method to deliver therapeutic agents intrathecally. PMID- 12895442 TI - Neurite elongation on chondroitin sulfate proteoglycans is characterized by axonal fasciculation. AB - In the developing or regenerating nervous system, migrating growth cones are exposed to regulatory molecules that positively and/or negatively affect guidance. Chondroitin sulfate proteoglycans (CSPGs) are complex macromolecules that are typically negative regulators of growth cone migration in vivo and in vitro. However, in certain cases, neurites sometimes traverse regions expressing relatively high levels of CSPGs, seemingly a paradox. In our continuing efforts to characterize CSPG inhibition in vitro, we manipulated the ratio of CSPGs to growth-promoting laminin-1 to produce a substratum that supports outgrowth of a subpopulation of dorsal root ganglia (DRG) neurites, while still being inhibitory to other populations of DRG neurons [Exp. Neurol. 109 (1990), 111; J. Neurobiol. 51 (2002), 285]. This model comprises a useful tool in the analysis of mechanisms of growth cone guidance and is particularly useful to analyze how CSPGs can be inhibitory under some conditions, and growth permissive under others. We grew embryonic (E9-10) chicken DRG neurons on nervous system-isolated, substratum bound CSPGs at a concentration that supports an intermittent pattern of outgrowth, alternating with regions adsorbed with growth-promoting laminin-1 alone, and analyzed outgrowth behaviors qualitatively and quantitatively. A novel finding of the study was that DRG neurites that elongated onto CSPGs were predominantly fasciculated, but immediately returned to a defasciculated state upon contact with laminin-1. Further, cursory inspection suggests that outgrowth onto CSPGs may be initially accomplished by pioneer axons, along which subsequent axons migrate. The outgrowth patterns characterized in vitro may accurately reflect outgrowth in vivo in locations where inhibitory CSPGs and growth promoting molecules are coexpressed, e.g., in the developing retina where fasciculated outgrowth may be instrumental in the guidance of retinal ganglion cells from the periphery to the optic fissure. PMID- 12895443 TI - Alterations on phosphodiesterase type 7 and 8 isozyme mRNA expression in Alzheimer's disease brains examined by in situ hybridization. AB - Phosphodiesterases (PDEs) play a central role in signal transduction by regulating intracellular levels of cyclic AMP (cAMP) and cGMP. It has been suggested that cAMP pathways could be upregulated in Alzheimer's disease. By in situ hybridization histochemistry we have determined the expression pattern of two newly described cAMP-specific phosphodiesterases families, PDE7 and PDE8, in several brain areas in control subjects. The hybridization levels of PDE8A mRNA were very low in all brain areas examined. High PDE7B and PDE8B mRNA signal intensities were found in the hippocampal formation. PDE7A was found to be present in both neuronal and non-neuronal cell populations. When the expression of these isozymes in control brains was compared with that in Alzheimer's disease brains staged according to Braak and Braak (Acta Neuropathol. (Berl.) 82 (1991), 239), we found that PDE8B was the only isozyme showing a significant increase, in cortical areas and parts of the hippocampal formation, at Braak stages III-VI. Our results show that the expression of specific cAMP PDE isoforms is selectively regulated in Alzheimer's disease and associated with the stages of the disease. The availability of animal models of Alzheimer's disease and of new pharmacological tools such as selective PDE inhibitors will allow study of the therapeutic potential of the control of cAMP levels in AD. PMID- 12895444 TI - BACE-2 is overexpressed in Down's syndrome. AB - Brain deposition of the amyloid-beta protein (Abeta) is a frequent complication of Down's syndrome (DS) patients. Abeta peptide is generated by endoproteolytic processing of Abeta precursor protein by gamma and beta secretases. Recently a transmembrane aspartyl protease, BACE, has been identified as the beta-secretase, and its homologous BACE-2 has also been described. BACE-2 gene resides on chromosome 21 in the obligate DS region. It cleaves Abeta precursor protein at its beta site and more efficiently at a different site within Abeta. In the present study we characterized the BACE-2 gene and protein expression in the DS patients and healthy control. We analyzed, by using a nonradioactive ribonuclease protection assay, the levels of BACE-2 mRNA expression in primary skin fibroblasts. The analysis revealed a 2.6-fold increase in BACE-2 mRNA levels in the DS group compared to the levels observed in the control group. Western blot analysis revealed no difference between DS and control in BACE-2 protein levels in the intracellular compartment. In the medium conditioned by fibroblast, we revealed an evident secretion of BACE-2 protein, represented by two different molecular weights, remarkably increased in DS fibroblasts. BACE-2 overexpression was also confirmed in the DS fetal brains and human neural embryonic DS stem cells in which conditioned media BACE-2 was secreted. These data highlight the importance of the extracellular compartment where BACE-2 overexpression could play a role in plaque formation in DS patients. PMID- 12895445 TI - Basal levels of metabolic activity are elevated in Genetic Absence Epilepsy Rats from Strasbourg (GAERS): measurement of regional activity of cytochrome oxidase and lactate dehydrogenase by histochemistry. AB - The Genetic Absence Epilepsy Rats from Strasbourg (GAERS) are considered an isomorphic, predictive, and homologous model of human generalized absence epilepsy. It is characterized by the expression of spike-and-wave discharges in the thalamus and cortex. In this strain, basal regional rates of cerebral glucose utilization measured by the quantitative autoradiographic [(14)C]2-deoxyglucose technique display a widespread consistent increase compared to a selected strain of genetically nonepileptic rats (NE). In order to verify whether these high rates of glucose metabolism are paralleled by elevated activities of the enzymes of the glycolytic and tricarboxylic acid cycle pathways, we measured by histochemistry the regional activity of the two key enzymes of glucose metabolism, lactate dehydrogenase (LDH) for the anaerobic pathway and cytochrome oxidase (CO) for the aerobic pathway coupled to oxidative phosphorylation. CO and LDH activities were significantly higher in GAERS than in NE rats in 24 and 28 of the 30 brain regions studied, respectively. The differences in CO and LDH activity between both strains were widespread, affected all brain systems studied, and ranged from 12 to 63%. The data of the present study confirm the generalized increase in cerebral glucose metabolism in GAERS, occurring both at the glycolytic and at the oxidative step. However, they still do not allow us to understand why the ubiquitous mutation(s) generates spike-and-wave discharges only in the thalamocortical circuit. PMID- 12895446 TI - Combination of dexanabinol and tempol in focal cerebral ischemia: is there a ceiling effect? AB - Because ischemic neuronal death is triggered by several parallel mechanisms, a combination of drugs active against individual death-promoting mechanisms may have synergistic effects. Dexanabinol is a noncompetitive NMDA antagonist with anti-inflammatory effects and tempol is a nitroxide antioxidant. Therefore, we explored whether their combined use results in smaller infarct volumes as compared with their individual administration. Rats underwent permanent middle cerebral artery occlusion (PMCAO) and were given vehicle, dexanabinol alone, tempol alone, or a combination of dexanabinol and tempol (n = 13 per group) 1 h later. Five animals in each group were evaluated with a motor rating scale 24 h after PMCAO and the infarct volumes were then measured. The remaining animals were examined with motor and behavioral scales up to 30 days after PMCAO and their infarct volumes were then determined. Motor disability and water maze latencies at all time points examined and infarct volumes at days 1 and 30 were significantly reduced in all active treatment groups when compared with vehicle. However, no significant differences were observed between the active treatment groups. In conclusions, combination therapy with dexanabinol and tempol does not appear to have additional neuroprotective effects compared to those conferred by each agent alone even when administered at optimal timing and dosing. Therefore, a ceiling neuroprotective effect that is impossible to overcome may exist. PMID- 12895447 TI - In the lithium-pilocarpine model of epilepsy, brain lesions are not linked to changes in blood-brain barrier permeability: an autoradiographic study in adult and developing rats. AB - Lithium-pilocarpine-induced status epilepticus (SE) leads to the genesis of massive neuronal loss in adult rats and to a lesser extent in P21 rats. Neuronal damage occurs mainly via a process of necrosis in limbic forebrain, cerebral cortex, thalamus, and substantia nigra. It is not known, however, whether damage is the result of local excitotoxic hyperactivity or if leakage at the blood-brain barrier (BBB) could participate in the damaging process. Therefore, we investigated the permeability of the BBB in adult and P21 rats using [alpha (14)C]aminoisobutyric acid, which does not cross an intact BBB, at 90 min after the onset of SE. At both ages, BBB opening occurred both in structures that will undergo damage (thalamus, septum, amygdala) and structures that will not be injured (globus pallidus, hypothalamus). In addition, neuronal damage occurs in the absence of increased BBB permeability in hippocampus, entorhinal cortex, and substantia nigra. Moreover, the increase in the intensity and distribution of BBB permeability changes is age-related, suggesting a differential activation of seizure circuits in adult and P21 rats. In summary, there is no clear correlation between the anatomical distribution of BBB opening and the occurrence of neuronal damage which, in this model, appears to rather depend on excitotoxic mechanisms due to major neuronal hyperexcitability. PMID- 12895448 TI - Neural trans-differentiation potential of hepatic oval cells in the neonatal mouse brain. AB - Although the existence of the hepatic oval cell (HOC), the liver stem cell has been known for almost 70 years, little is known about the potential for this adult stem cell to trans-differentiate into cells of other tissues. While their origin remains enigmatic, HOCs share many similarities with hematopoietic stem cells. Recent studies have revealed that a small percentage of HOCs can arise from a bone marrow-derived stem cell source. Here we report that, like bone marrow stem cells, HOCs can survive transplantation to the neonatal mouse brain and show signs of trans-differentiation by adopting the morphology and antigenic phenotype of both macro- and microglia cells. Trans-differentiated microglia cells were functional, showing active phagocytosis when cotransplanted with latex microbeads in vivo. In addition to glial markers, a small number of transplanted HOCs were immunopositive for neuronal markers, but displayed ambiguous phenotype, making their characterization difficult. PMID- 12895449 TI - Mats made from fibronectin support oriented growth of axons in the damaged spinal cord of the adult rat. AB - A variety of biological as well as synthetic implants have been used to attempt to promote regeneration into the damaged spinal cord. We have implanted mats made from fibronectin (FN) into the damaged spinal cord to determine their effectiveness as a substrate for regeneration of axons. These mats contain oriented pores and can take up and release growth factors. Lesion cavities 1 mm in width and depth and 2 mm in length were created on one side of the spinal cord of adult rats. FN mats containing neurotrophins or saline were placed into the lesion. Mats were well integrated into surrounding tissue and showed robust well oriented growth of calcitonin gene-related peptide, substance P, GABAergic, cholinergic, glutamatergic, and noradrenergic axons into FN mats. Transganglionic tracing using cholera toxin B indicated large-diameter primary afferents had grown into FN implants. Schwann cells had also infiltrated FN mats. Electron microscopy confirmed the presence of axons within implants sites, with most axons either ensheathed or myelinated by Schwann cells. Mats incubated in brain-derived neurotrophic factor and neurotrophin-3 showed significantly more neurofilament positive and glutamatergic fibers compared to saline- and nerve growth factor incubated mats, while mats incubated with nerve growth factor showed more calcitonin gene-related peptide-positive axons. In contrast, neurotrophin treatment had no effect on PGP 9.5-positive axons. In addition, in some animals with neurotrophin-3-incubated mats, cholera toxin B-labelled fibers had grown from the mat into adjoining intact areas of spinal cord. The results indicate that FN mats provide a substrate that is permissive for robust oriented axonal growth in the damaged spinal cord, and that this growth is supported by Schwann cells. PMID- 12895450 TI - The chondroitin sulfate proteoglycans neurocan, brevican, phosphacan, and versican are differentially regulated following spinal cord injury. AB - Chondroitin sulfate proteoglycans (CSPGs) are extracellular matrix (ECM) molecules that are widely expressed throughout the developing and adult CNS. In vitro studies demonstrate their potential to restrict neurite outgrowth, and it is believed that CSPGs also inhibit axonal regeneration after CNS injury in vivo. Previous studies demonstrated that CSPGs are generally upregulated after spinal cord injury, and more recent reports have begun to identify individual proteoglycans that may play dominant roles in limiting axonal regeneration. The current study systematically examined the extended deposition patterns after CNS injury of four putatively inhibitory CSPGs that have not been extensively investigated previously in vivo: neurocan, brevican, phosphacan, and versican. After spinal cord injury, neurocan, brevican, and versican immunolabeling increased within days in injured spinal cord parenchyma surrounding the lesion site and peaked at 2 weeks. Neurocan and versican were persistently elevated for 4 weeks postinjury, and brevican expression persisted for at least 2 months. On the other hand, phosphacan immunolabeling decreased in the same region immediately following injury but later recovered and then peaked after 2 months. Combined glial fibrillary acidic protein (GFAP) immunohistochemistry and in situ hybridization demonstrated that GFAP astrocytes constituted a source of neurocan production after spinal cord injury. Thus, the production of several CSPG family members is differentially affected by spinal cord injury, overall establishing a CSPG-rich matrix that persists for up to 2 months following injury. Optimization of strategies to reduce CSPG expression to enhance regeneration may need to target several different family members over an extended period following injury. PMID- 12895451 TI - Early mitochondrial dysfunction occurs in motor cortex and spinal cord at the onset of disease in the Wobbler mouse. AB - The Wobbler mouse is recognized as an animal model for motoneuron disease that exhibits motoneuron pathology. We have recently demonstrated the occurrence of mitochondrial dysfunction in the Wobbler mouse brain. The aim of the present study was to evaluate whether mitochondrial dysfunction occurred at an early age at the time where disease symptoms appear, and whether it was more pronounced in the motor cortex or in the spinal cord. We report here a significant decrease in mitochondrial state 3 and 4 respiration rates at an early age in the Wobbler spinal cord. In addition, there was a pronounced decrease in oxidative phosphorylation in mitochondria isolated from both spinal cord and motor cortex in both age groups. This mitochondrial dysfunction was accompanied by impairment of complex I activity in mitochondria isolated from spinal cord at an early age. Decreases in complex III and IV activities were observed only in mitochondria isolated from the motor cortex at an early age, but impairment of complex III activity prevailed until later in the disease. We conclude that mitochondrial dysfunction ensues at an early stage of the disease and is more pronounced in the spinal cord, which correlates with previous studies that reported degeneration of spinal cord motorneurons. PMID- 12895452 TI - Decreased platelet cytochrome c oxidase activity is accompanied by increased blood lactate concentration during exercise in patients with Alzheimer disease. AB - Increasing evidence indicates that mitochondrial dysfunction occurs in the central nervous system as well as in the peripheral tissues from Alzheimer's disease (AD) patients. We have recently shown that mitochondrial cytochrome c oxidase (COX) activity is significantly reduced in brain and platelets from AD patients compared to controls. In the present study we investigated whether impaired COX activity could have functional consequences on energy metabolism. Blood lactate concentration was monitored at rest and during incremental exercise in 22 AD patients in whom COX activity in platelets was decreased compared to controls (35.7 +/- 11.4 vs 48.4 +/- 1.4 nmol/min/mg, P < 0.01). In both resting and exercising conditions, blood lactate was significantly higher in AD patients than in controls. Although the magnitude of exercise-related lactate accumulation was not different between the two groups, an anticipated anaerobic lactate threshold during the incremental forearm exercise was found in AD patients (50% of maximal voluntary contraction MVC compared to 60% in controls). COX activity was inversely related to lactate at a significant level for resting condition (r = -0.65) and borderline for anaerobic threshold exercise level. These results support the hypothesis of a systemic impairment of the mitochondrial function in AD and indicate that decreased COX activity could have functional consequences on metabolism. PMID- 12895453 TI - Gabapentin reverses mechanical allodynia induced by sciatic nerve ischemia and formalin-induced nociception in mice. AB - The anticonvulsant drug gabapentin has been demonstrated to alleviate symptoms of painful diabetic neuropathy as well as other types of neuropathic pain. The aim of the present study was to investigate the effect of gabapentin in a recently developed mouse model of peripheral neuropathy. This model is based on a photochemical ischemic lesion of the sciatic nerve generated by laser-induced activation of the photosensitizing dye erythrosin B. Following laser irradiation of the sciatic nerve for 2, 5, or 10 min, tactile allodynia was observed during at least 3 weeks. The degree of allodynia was most marked following 10 min of irradiation. Subcutaneous administration of gabapentin [175-300 micromol/kg ( approximately 30-51 mg/kg), cumulative doses, at 1-h intervals] significantly reversed tactile allodynia induced by 10-min laser irradiation. The maximal dose of gabapentin increased the withdrawal threshold from approximately 0.55 to approximately 1.85 g (i.e., about 77% of the threshold in normal animals, approximately 2.4 g). Gabapentin did not affect the tactile withdrawal threshold in intact animals. A dose of gabapentin (100 micromol/kg, sc) that had no effect on allodynia was found to significantly reduce the pain behavior during phase 2 of the formalin test. The present study demonstrates that systemic administration of gabapentin suppresses both allodynia induced by an ischemic lesion of the sciatic nerve and pain behavior in the formalin test. PMID- 12895454 TI - Angiogenic and neurotrophic effects of vascular endothelial growth factor (VEGF165): studies of grafted and cultured embryonic ventral mesencephalic cells. AB - The present series of experiments investigated the effects of vascular endothelial growth factor (VEGF165) on adult rat striatal cerebrovasculature and embryonic dopamine (DA) neuron allografts in a rat model of Parkinson's disease (PD). We examined VEGF165's ability to (1) alter the vascular network of the adult rat striatum, (2) influence the vascular growth of solid embryonic day 14 (E14) ventral mesencephalic (VM) grafts when placed into a VEGF-pretreated host striatum, (3) alter the function and survival of E14 VM grafts when transplanted into an adult DA-deleted striatum, and (4) influence cell survival and neurite growth in cultures of E14 VM cells. We demonstrate here that a single bolus injection of VEGF165 into the adult rat striatum significantly increases the amount of vasculature in the vicinity of the injection site in a delayed and transient manner when compared to saline controls. Transplanting solid E14 VM grafts into the VEGF165-pretreated striatum resulted in a homogeneous distribution of small blood vessels throughout the graft, a pattern that closely resembles mature adult vasculature. In contrast, grafts in the control condition contained a patchy distribution of heavily dilated vessels. Behavioral measurements indicate that VEGF pretreatment of the intrastriatal graft site accelerates recovery of amphetamine-induced rotational asymmetry in unilateral 6 OHDA lesioned rats. Unexpectedly, however, VEGF pretreatments failed to increase survival of tyrosine hydroxylase-immunoreactive (THir) neurons in the grafts. In contrast to this finding in vivo, adding VEGF165 to glial-reduced E14 rat VM cultures produced a fourfold increase in THir cell survival and a doubling in the length of THir neurites. We conclude that with the proper method of delivery, VEGF165 may prove to be one of several strategies necessary to significantly improve the survival and function of fetal VM tissue grafts. PMID- 12895455 TI - Neutrophils do not contribute to infarction, oxidative stress, and NO synthase activity in severe brain ischemia. AB - Polymorphonuclear leukocytes (PMNs) were reported to contribute to ischemia reperfusion-induced brain damage. The present work examined whether PMN infiltration is deleterious in a severe model of transient focal cerebral ischemia and in which part PMNs contribute to oxidative stress and nitric oxide (NO) production. A 20-min occlusion of the left middle cerebral artery and both common carotid arteries was performed in rats. Infarction was maximal 24 h after reperfusion, while accumulation of PMNs in infarcted tissue was not significant before 48 h. Moreover, neutropenia induced by vinblastine (0.5 mg/kg iv) significantly decreased by 60-80% PMN infiltration 48 h after reperfusion but did not reduce the infarct volume. Thus PMNs do not contribute to cerebral injury in our model. Furthermore, decreased PMN infiltration modified neither oxidative stress evaluated by glutathione concentrations nor NO synthase activities 48 h after reperfusion. In conclusion, our results suggest that PMNs are not involved in severe cerebral ischemia and that anti-PMN strategies may be inefficient in some pathological conditions. PMID- 12895456 TI - Ammonia potentiates methylmalonic acid-induced convulsions and TBARS production. AB - Hyperammonemia is a common finding in children with methylmalonic acidemia, an inherited metabolic disease characterized by mental retardation, convulsions, and accumulation of methylmalonic acid (MMA). Although it has been suggested that MMA induces convulsions through succinate dehydrogenase (SDH) inhibition, very little is known about the contribution of hyperammonemia to the development of convulsions in these patients. In the present study we investigated the effects of ammonium ions on the convulsant action of MMA, MMA-induced inhibition of striatal succinate dehydrogenase, and the striatal content of thiobarbituric acid reactive substances (TBARS). Adult rats were injected with ammonium acetate (1.5 mmol/kg, sc) or sodium acetate (1.5 mmol/kg, sc), followed 5 min later by buffered MMA (3 micromol/microl) or NaCl (4.5 micromol/microl) injected into the striatum. The animals were observed in an open field for the appearance of convulsive episodes. After 30 min of behavioral evaluation, the animals were sacrificed and had their striatal TBARS content measured. Ammonium acetate pretreatment caused no behavioral effects per se, but potentiated MMA-induced convulsions and increased basal TBARS content and MMA-induced TBARS production in the striatum. Ammonium chloride had no effect on basal succinate dehydrogenase activity and did not alter MMA-induced inhibition of SDH in vitro. These results suggest that ammonia potentiates MMA-induced behavioral effects through a mechanism that does not involve further succinate dehydrogenase inhibition, but may involve facilitation of MMA-induced oxidative damage and provide evidence that ammonia and MMA may have mutually additive toxicity. PMID- 12895457 TI - Lamotrigine and remacemide protect striatal neurons against in vitro ischemia: an electrophysiological study. AB - In the present study, we investigated the cellular and synaptic mechanisms underlying the neuroprotective action of lamotrigine and remacemide. Both drugs, in fact, have been reported to exert a neuroprotective action in in vivo animal models of ischemia. To address this issue, electrophysiological recordings and cell swelling measurements were performed from striatal neurons in control condition and during combined oxygen and glucose deprivation (in vitro ischemia) in a brain slice preparation. Lamotrigine, remacemide, and the active desglycinyl metabolite of remacemide, D-REMA, induced a concentration-dependent reduction of both repetitive firing discharge and excitatory postsynaptic potentials. However, while remacemide and D-REMA exerted their inhibitory action on glutamatergic transmission by blocking NMDA receptors, lamotrigine exerted a preferential presynaptic action, as indicated by its ability to increase paired-pulse facilitation. Both remacemide and lamotrigine were found to be neuroprotective against the irreversible field potential loss and cell swelling induced by in vitro ischemia, and coadministration of low concentrations of these drugs exerted an additive neuroprotective action. A combined use of lamotrigine and remacemide could be employed in clinical trials to enhance neuroprotection in neurological disorders involving an abnormal striatal glutamatergic transmission. PMID- 12895458 TI - Two organizational effects of pubertal testosterone in male rats: transient social memory and a shift away from long-term potentiation following a tetanus in hippocampal CA1. AB - The organizational role of pubertal androgen receptor (AR) activation in synaptic plasticity in hippocampal CA1 and in social memory was assessed. Earlier data suggest pubertal testosterone reduces adult hippocampal synaptic plasticity. Four groups were created following gonadectomy at the onset of puberty: rats given testosterone; rats given testosterone but with the AR antagonist flutamide, present during puberty; rats given testosterone at the end of puberty; and rats given cholesterol at the end of puberty. A tetanus normally inducing long-term potentiation (LTP) was used to stimulate CA1 in the urethane-anesthetized adults during the dark phase of their cycle. Social memory was assessed prior to electrophysiology. Social memory for a juvenile rat at 120 min was seen only in rats not exposed to AR activation during puberty. Pubertal AR activation may induce the reduced social memory of male rats. Early CA1 LTP occurred following tetanus in rats with no pubertal testosterone. Short-term potentiation occurred in rats exposed to pubertal testosterone. Unexpectedly, rats with pubertal AR activation developed long-term depression (LTD). The same pattern was seen in normal male rats. Lack of LTP during the dark phase is consistent with other data on circadian modulation of CA1 LTP. No correlations were seen among social memory scores and CA1 plasticity measures. These data argue for two organizational effects of pubertal testosterone: (1) CA1 synaptic plasticity shifts away from potentiation toward depression; (2) social memory is reduced. Enduring effects of pubertal androgen on limbic circuits may contribute to reorganized behaviors in the postpubertal period. PMID- 12895459 TI - Long-term effects of clomethiazole in a model of global ischemia. AB - The failure of neuroprotective drugs in clinical trials has raised questions about the predictive value of animal models. To address this issue we reexamined the efficacy of clomethiazole using functional and histological outcome measures in combination with long-term survival times. Gerbils were exposed to 5 min of global ischemia and received 400 mg/ml clomethiazole (via osmotic minipump) plus a bolus injection (60 mg/kg) 30 min after ischemia. Brain temperature was maintained at approximately 36.5 degrees C during ischemia and for the first 30 min after ischemia, and was monitored in all groups for 24 h. Subgroups of clomethiazole-treated gerbils had their temperatures regulated in the normothermic range while in other animals temperature was not controlled. Open field habituation tests were conducted 5, 10, 30, and 60 days after occlusion. CA1 cell counts and CA1 slice recordings were done at the conclusion of behavioral testing. Clomethiazole significantly attenuated CA1 cell loss at 10-, 30-, and 60-day survival. A modest reduction in habituation deficits was evident only on Day 10 (P < 0.05). Similarly, field potential amplitude was not maintained in the rostral CA1 region. Clomethiazole produced mild hypothermia that developed over several hours. Based on short-term CA1 cell counts, clomethiazole provided significant histological protection with limited functional preservation. Neuroprotection disappeared when longer survival times (60 day) were employed and temperature confounds eliminated. These data demonstrate the necessity of utilizing more clinically relevant survival times and carefully monitoring/regulating postischemic temperature when assessing potential neuroprotective compounds. PMID- 12895460 TI - Dopamine release rather than content in the caudate putamen is associated with behavioral changes in the iron rat model of Parkinson's disease. AB - The effects of intranigral iron injection on dopamine (DA) release and content in the caudate putamen (CPu) and their relationship to DA-related behavioral response were investigated in rats. Different concentrations of FeCl(3) (10, 20, and 40 microg) and saline were injected separately into the left substantia nigra. In some experiments, rats were pretreated with desferrioxamine or saline before iron injection. After 3 weeks, changes in behavioral response, DA release, and DA content in the CPu were determined. In all iron injection groups (10, 20, and 40 microg), DA content in the lesioned side of the brain was significantly decreased, showing a significant linear correlation (R(2) = 0.981, P = 0.01), and DA turnover ratio significantly increased (both P = 0.01, 0.01 and 0.001 vs unlesioned sides, respectively). However, injection dosages of 10 or 20 microg of iron did not lead to significant changes in DA release in the CPu or in behavioral response. At the 40-microg dosage, it was found that DA release in the lesioned side and rearing activity both were significantly reduced (all P = 0.01 vs unlesioned side or control) and apomorphine-induced rotation was observed. Pretreatment with desferrioxamine significantly inhibited the effect of iron on DA release and content. These results demonstrate that iron injection can damage dopaminergic neurons and suggest that DA release, rather than DA content, in the CPu is associated with DA-related behavioral changes in this PD model. PMID- 12895461 TI - Antidyskinetic effect of magnesium sulfate in MPTP-lesioned monkeys. AB - The antiparkinsonian action of an NMDA receptor antagonist, magnesium sulfate (50, 100, and 200 mg/kg), alone and in association with levodopa was explored in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-lesioned parkinsonian and control rhesus monkeys. At the three doses tested, magnesium sulfate decreased levodopa-induced dyskinesia [cumulative dyskinetic scores after levodopa: 129 +/- 13; after levodopa and magnesium sulfate: 65 +/- 14 (50 mg/kg), P < 0.001; 64 +/- 10 (100 mg/kg), P < 0.001; 66 +/- 21 (200 mg/kg), P < 0.001, compared to levodopa administration alone]. These results show that magnesium sulfate importantly reduces levodopa-induced dyskinesia. PMID- 12895462 TI - Effects of stimulus frequency and age on bidirectional synaptic plasticity in the dentate gyrus of freely moving rats. AB - We investigated the frequency-dependent transition from homosynaptic long-term depression (LTD) to long-term potentiation (LTP) at the lateral perforant pathway/dentate gyrus synapse in adult (90 days of age) and immature (15 days of age) awake, freely moving rats. Dentate-evoked field potentials were recorded and analyzed using the population spike amplitude and the field EPSP slope measures following sustained stimulation (900 pulses) of the lateral perforant pathway at various frequencies (1, 3, 7, 30, 50, or 200 Hz). Our results indicate that both the strength and the direction (LTP or LTD) of synaptic plasticity vary as a function of activation frequency: sustained low-frequency stimulation ranging from 1 to 7 Hz results in depression of activated synapses, whereas high frequency stimulation (30-200 Hz) produces potentiation. In addition, a significant (P < 0.01) ontogenetic shift in the frequency of transition from LTD to LTP was observed; the transition frequency in immature animals was significantly lower than that obtained in adult animals. These observations agree strongly with the prediction of the Bienenstock-Cooper-Munro theory of synapse modification, indicating perhaps a neurophysiological basis for this theoretical model of learning in the dentate gyrus of awake behaving rats. PMID- 12895463 TI - The effect of acute hypoxemia and hypotension on adenosine-mediated depression of evoked hippocampal synaptic transmission. AB - The present study was designed to investigate the relative contributions of arterial P(O(2)), local cerebral blood flow, and oxygen delivery to the adenosine A(1) receptor-mediated depression of evoked synaptic transmission recorded in the rat hippocampus. Urethane-anesthetized rats were given a unilateral common carotid artery occlusion and then placed in a stereotaxic apparatus for stimulation and recording of bilateral hippocampal field excitatory postsynaptic potentials (fEPSPs). Arterial blood gases, mean arterial blood pressure (MAP), and bilateral hippocampal blood flow (HBF) were also measured. Arterial P(O(2)), HBF, and oxygen delivery were manipulated using normoxic hypotension, hypoxic hypotension, and hypoxic normotension. Both hypoxic hypotension and normoxic hypotension resulted in decreased HBF, decreased oxygen delivery, and a depression of the evoked fEPSP limited to the hippocampus ipsilateral to the occlusion. The enhanced HBF and oxygen delivery associated with increased MAP resulted in a restoration and maintenance of hippocampal fEPSPs despite sustained hypoxemia. The adenosine A(1) receptor-mediated depression of the fEPSP was more strongly correlated with changes in HBF and oxygen delivery than with arterial P(O(2)). We propose that adenosine plays an important role mediating the depression of neuronal activity associated with reduced oxygen delivery characteristically observed in ischemic brain tissue. PMID- 12895464 TI - Mechanistic considerations in high-throughput screening. PMID- 12895465 TI - Release of the self-quenching of fluorescence near silver metallic surfaces. AB - Fluorescein is one of the most widely used fluorescent probes in microscopy, biotechnology, and clinical assays. One difficulty with fluorescein is its self quenching, which results in decreased intensities with increasing labeling density. In this study we examined human serum albumin (HSA), which contained one to nine covalently linked fluorescein molecules per molecule of HSA. The occurrence of homo resonance energy transfer for labeling ratios greater than 1 were confirmed by decreases in the relative quantum yields, anisotropies, and lifetimes. We found that most of the self-quenching can be partially eliminated by proximity of the labeled protein to metallic silver particles. These results suggest the use of heavily labeled proteins and metallic colloids to obtain ultrabright reagents for use in immunoassays, imaging, and other applications. PMID- 12895466 TI - Essential cysteine-alkylation strategies to monitor structurally altered estrogen receptor as found in oxidant-stressed breast cancers. AB - Oxidant-induced structural modifications within the cysteine-rich DNA-binding domain (DBD) of the overexpressed estrogen receptor (ER) likely contribute to its loss of DNA-binding function and altered transcriptional activity during human breast cancer development. Using recombinant ER protein as a model, procedures to detect such endogenously produced structural changes in the two Cys(4)-type zinc fingers within the DBD of ER extracted from breast cancer cells are being developed. Unfortunately, ex vivo oxidation of these ER-DBD cysteine residues can occur during routine ER purification and preparation procedures. Also, cysteine residues readily undergo thiol-disulfide exchange reactions that can result in artificial oxidation and incorrect disulfide bond assignments. These problems can be circumvented by an initial irreversible alkylation of all free thiols followed by reduction of any disulfides and treatment with a second alkylating agent, prior to proteolysis and high-performance liquid chromatography mass spectrometry analysis of peptides in the doubly alkylated ER digest, to differentiate between the originally free and the disulfide-bonded cysteine residues. Although the use of chemically identical but isotopically different alkylating agents was more effective than the use of chemically different alkylating agents, subsequent problems were encountered with incomplete alkylation of particular Cys residues in the native ER protein. To overcome this limitation, the initial alkylation was accompanied by denaturation and the second alkylation was carried out during the proteolytic digestion. These improved analytical strategies should facilitate the monitoring of structurally altered endogenous ER produced within oxidant-stressed human breast cancer cells. PMID- 12895467 TI - Cobalt(II) salophen-modified carbon-paste electrode for potentiometric and voltammetric determination of cysteine. AB - A chemically modified electrode constructed by incorporating N,N(') bis(salicylidene)-1,2-phenylenediaminocobalt(II) into carbon-paste matrix was used as a sensitive electrochemical sensor for detection of cysteine. The resulting electrode exhibits catalytic properties for the electrooxidation of cysteine and lowers the overpotential for the oxidation of this compound. The faster rate of electron transfer results in a near-Nernstian behavior of the modified electrode and makes it a suitable potentiometric and voltammetric sensor for the fast and easy determination of cysteine. A linear response in concentration range from approximately 2 microM to 0.01 M was obtained with a detection limit of 1 microM for the potentiometric detection of cysteine. The modified electrode was also used for the amperometric and differential pulse voltammetric determination of cysteine and the results were compared with those of the potentiometric method. PMID- 12895468 TI - Spectropotentiometric analysis of metal binding to structural zinc-binding sites: accounting quantitatively for pH and metal ion buffering effects. AB - Studies of the metal-binding affinity of protein sites are ubiquitous in bioinorganic chemistry and are valuable for the information that they can provide about metal speciation and exchange in biological systems. The potential for error in these studies is high, however, since many competing equilibria are present in solution and must be taken into consideration. Here, we report a new spectropotentiometric titration apparatus that allows pH and UV-vis absorption to be monitored simultaneously on small samples under inert atmosphere. In addition, we explain how data obtained from the complex equilibria can be combined with tabulated information about the protonation and metal-binding constants for common buffers to provide detailed, quantitative information about metal-protein interactions. Application of this approach to the investigation of metal binding to structural zinc-binding domains and common pitfalls encountered when performing these experiments are also discussed. We have used this approach to reevaluate the metal-binding constants of the N-terminal zinc-binding peptide from the HIV-1 nucleocapsid protein (10(-8)M2)-beta-D-galactopyranosyl-(1-->2)-beta-D-glucuronopyranosyl 21-O-alpha-L-arabinopyranoside and 3beta,21alpha,22beta,24-tetrahydroxyolean-12 en-3-O-beta-D-galactopyranosyl-(1-->2)-beta-D-glucuronopyranosyl-21-O-alpha-L rhamnopyranoside, along with eight other saponins and one triterpene previously reported from other legumes, were isolated from the aerial parts of Lupinus oreophilus collected in northern Chile. The structures of the isolated compounds were established with the help of extensive spectroscopic techniques. PMID- 12895530 TI - Diterpenoid from Salvia greggii. AB - The structure of a diterpenoid, designated salvigresin, that was isolated from the aerial parts of Salvia greggii, has been confirmed by spectroscopic investigation and X-ray analysis. PMID- 12895531 TI - Germacranolides from seeds of the endangered Umbelliferae species Rouya polygama. AB - From the etheral extract of Rouya polygama Coincy seeds (Umbelliferae) six new germacranolides 1-6 were isolated. The structures were elucidated by chemical methods and spectroscopic analysis. Compounds 1 and 4 exhibited activity against Artemia salina larvae. PMID- 12895532 TI - A trinorsesterterpene glycoside from the North American fern Woodwardia virginica (L.) Smith. AB - A new trinorsesterterpene glycoside was isolated from the ethanol extract of the American fern Woodwardia virginica having a 3-[6-(4,8-dimethyl-nona-1,3,7 trienyl)-4-hydroxy-2,6-dimethyl-cyclohex-1-enyl]-3-hydroxypropionic acid, as the aglycone and a saccharide moiety linked at C-4 to glucoses, xylose or arabinofuranose. The structure was elucidated using extensive spectroscopic analysis (1D and 2D NMR, MS, IR and UV) including determination of absolute stereochemistry by means of the MTPA and PGME derivatives and also by chemical methods. PMID- 12895533 TI - Guaiane-type sesquiterpenoids from Alisma orientalis. AB - Two guaiane-type sesquiterpenoids named orientalol E (1) and orientalol F (3) were isolated from the rhizome of Alisma orientalis (SAM) JUZEP together with two known guaiane-type sesquiterpenoids alismol (2) and alismoxide (4). Their relative stereostructures were elucidated by spectroscopic methods, whereas absolute stereostructures were determined on the basis of chemical correlation. PMID- 12895534 TI - Mulinane-type diterpenoids from Mulinum spinosum. AB - In addition to the known mulinolic and mulinenic acids, two diterpenoids, mulin 11-ene-13-alpha,14-alpha-dihydroxy-20-oic and mulin-12-ene-14-one-20-oic acids, were isolated from the aerial parts of Mulinum spinosum (Cav.) Pers. (Apiaceae). Their structures were determined based on spectroscopic studies. PMID- 12895535 TI - Iridoids from Kigelia pinnata DC. fruits. AB - From the fruits of Kigelia pinnata DC., a new furanone derivative formulated as 3 (2'-hydroxyethyl)-5-(2"-hydroxypropyl)-dihydrofuran-2(3H)-one (1), and four new iridoids named; 7-hydroxy viteoid II (2), 7-hydroxy eucommic acid (3), 7-hydroxy 10-deoxyeucommiol (4) and 10-deoxyeucommiol (5) have been isolated together with seven known iridoids, jiofuran (6), jioglutolide (7), 1-dehydroxy-3,4 dihydroaucubigenin (8), des-p-hydroxybenzoyl kisasagenol B (9), ajugol (10), verminoside (11) and 6-trans-caffeoyl ajugol (12). The structures of the isolated compounds were characterized by different spectroscopic methods. PMID- 12895536 TI - Isolation and structure elucidation of flavonoid and phenolic acid glycosides from pericarp of hot pepper fruit Capsicum annuum L. AB - Hot pepper fruits (Capsicum annuum L.) var. Bronowicka Ostra have been studied with regard to content of flavonoids and other phenolics. Nine compounds were isolated from pericarp of pepper fruits by preparative HPLC. Their structures were identified by chromatographic (analytical HPLC) and spectroscopic (UV, NMR) techniques. Two of the identified compounds, trans-p-ferulylalcohol-4-O-(6-(2 methyl-3-hydroxypropionyl) glucopyranoside and luteolin-7-O-(2-apiofuranosyl-4 glucopyranosyl-6-malonyl)-glucopyranoside were found for the first time in the plant kingdom. Additionally compounds: trans-p-feruloyl-beta-D-glucopyranoside, trans-p-sinapoyl-beta- D-glucopyranoside, quercetin 3-O-alpha-L-rhamnopyranoside 7-O-beta-D-glucopyranoside, luteolin 6-C-beta-D-glucopyranoside-8-C-alpha-L arabinopyranoside, apigenin 6-C-beta-D-glucopyranoside-8-C-alpha-L arabinopyranoside and luteolin 7-O-[2-(beta-D-apiofuranosyl)-beta-D glucopyranoside] were found for the first time in pepper fruit Capsicum annuum L. PMID- 12895537 TI - A dehydrotrimer of ferulic acid from maize bran. AB - A new phenolic acid trimer was detected by coupled liquid chromatography/mass spectroscopy in alkali extracts of maize bran. The trimer was purified by preparative silica gel chromatography. The structure of the new compound was elucidated on the basis of 1D and 2D NMR and corresponded to a 4-O-8', 5'-5" dehydrotriferulic acid. PMID- 12895538 TI - Caffeoyl sugar esters and an ellagitannin from Rubus sanctus. AB - The new natural caffeoyl esters, 3,6-di-O-caffeoyl-(alpha/beta)-glucose and 1-O caffeoyl-beta-xylose, together with the hitherto unknown natural tannin, 2,3-O hexahydroxydiphenoyl-4,6-O-sanguisorboyl-(alpha/beta)-glucose, have been isolated from the aqueous alcohol aerial part extract of Rubus sanctus. Establishment of all structures was based on the chemical and spectral evidence, including ESI-MS and 2D NMR. PMID- 12895539 TI - Two oligostilbenes, cis- and trans-diptoindonesin B, from Dryobalanops oblongifolia. AB - Two oligostilbenes, cis- and trans-diptoindonesin B, have been isolated from the tree bark of Dryobalanops oblongifolia (Dipterocarpaceae). The structures and relative configurations of both compounds were determined on the basis of spectroscopic evidence, including 2D-NMR spectroscopic analysis. PMID- 12895540 TI - Thelephantins D-H: five p-terphenyl derivatives from the inedible mushroom Thelephora aurantiotincta. AB - Five p-terphenyl derivatives named thelephantins D-H (1-5) together with nine known compounds, thelephantins A-C (6-8), ganbajunin E (9), p-hydroxylbenzoic acid (10), ganbajunin C (11), thelephorin A (12), 2-O-methylatromentin (13) and atromentin (14), were isolated from the methanolic extract of fruit bodies of the Thelephoraceous Basidiomycete Thelephora aurantiotincta. Their structures were elucidated by high-resolution MS, 2D NMR, IR and UV spectroscopic analysis. PMID- 12895541 TI - 1-Methyl anthraquinones and their biogenetic precursors from Stereospermum personatum. AB - Two novel 1(17)-methyl anthraquinones, sterequinone-A and -D, their biogenetic precursors sterequinone-B, -C, and a new naphthoquinone sterequinone-E along with a known naphthoquinone, sterekunthal-B, have been isolated from the petroleum ether extract of stem bark of Stereospermum personatum. Their structures were established using spectroscopic methods. PMID- 12895542 TI - Two cyclohexenone glycosides from the North American fern Woodwardia virginica (L.) Smith. AB - New glycosides having multisubstituted cyclohex-2-enones as aglycones and saccharide moieties consisting of three and four glucoses, respectively were isolated from the ethanol extract of the American fern Woodwardia virginica. The structures were elucidated using extensive spectroscopic analysis (1D and 2D NMR, MS, IR, CD and UV) including determination of absolute stereochemistry by chemical methods. PMID- 12895543 TI - Benzofurans and another constituent from seeds of Styrax officinalis. AB - The benzofuran constituents of the seeds of Styrax officinalis were investigated. From the hexane extract, two new constituents named 5-(3"benzoyloxypropyl)-7 methoxy-2-(3',4'-methylenedioxyphenyl)-benzofuran (5) and 4-[3"-(1c methylbutanoyloxy)propyl]-2-methoxy-(3',4'-methylenedioxyphenyl)-1a, 5b dihydrobenzo-[3,4]-cyclobutaoxirene (6) were isolated together with four known compounds, 5-[3"-(1c-methylbutanoyloxy)propyl]-7-methoxy-2-(3',4' dimethoxyphenyl)-benzofuran (4), 5-[3"-(1c-methylbutanoyloxy)propyl]-7- methoxy-2 (3',4'-methylenedioxyphenyl)-benzofuran (3), 5-(3"-acetoxypropyl)-7-methoxy2 (3',4'-methylenedioxphenyl)-benzofuran (2) and 5-(3"-hydroxypropyl)-7-methoxy-2 (3',4'-met hylenedioxyphenyl)-benzofuran (1). Although the compounds 1, 2, and 3 have been isolated previously from the seeds of Styrax obassia, this is the first record of their isolation from seeds of Styrax officinalis. The structures of the isolated compounds were established by 1D- and 2D-NMR (HMBC, HMQC, COSY), FABMS and high-resolution ESI FTMS. PMID- 12895544 TI - Polypropionate lactones of deoxysugars glycosides from slime mold Lycogala epidendrum. AB - Two novel polypropionate lactone glycosides (1 and 2, i.e. lycogalinosides A and B) were isolated from the slime mold Lycogala epidendrum. Their structures, including the absolute configurations of the hydroxyl and methyls groups, were determined by means of extensive spectroscopic data such as mass, IR, UV, and 1D and 2D NMR spectra and chemical degradation followed by spectroscopic and chromatographic analysis. Compounds 1 and 2 are unique in structure containing a 2-deoxy-alpha-L-fucopyranosyl-(1-4)-6-deoxy-beta-D-gulopyranosyl unit and a beta D-olivopyranosyl-(1-4)-beta-D-fucopyranosyl unit, respectively, and showed growth inhibitory activities against Gram-positive bacteria. PMID- 12895545 TI - Diterpene esters of aristolochic acids from Aristolochia pubescens. AB - From the acetone and ethanol extracts of the tubercula of Aristolochia pubescens, two diterpene esters of aristolochic acids were isolated, together with 23 known compounds. The structures of the compounds were determined on the basis of spectroscopic analysis. PMID- 12895546 TI - Glucosides from Vitex agnus-castus. AB - The methanolic extract of the flowering stems of Vitex agnus-castus yielded three new iridoids: 6'-O-foliamenthoylmussaenosidic acid (agnucastoside A), 6'-O-(6,7 dihydrofoliamenthoyl)mussaenosidic acid (agnucastoside B) and 7-O-trans-p coumaroyl-6'-O-trans-caffeoyl-8-epiloganic acid (agnucastoside C) in addition to four known iridoids (aucubin, agnuside, mussaenosidic acid and 6'-O-p hydroxybenzoylmussaenosidic acid) and one known phenylbutanone glucoside (myzodendrone). The structure elucidations were mainly done by spectroscopic methods (1D and 2D NMR spectra) and MS data interpretation. The purified compounds were tested for biological activities against various microorganisms and cancer cell lines. PMID- 12895547 TI - Sulfur containing derivatives from Ferula persica var. latisecta. AB - Two new sulfur containing derivatives, t-butyl 3-[(1-methylpropyl)dithio]-2 propenyl malonate (1), t-butyl 3-[(1-methylthiopropyl)thio]-2-propenyl malonate (2) were isolated from the roots of Ferula persica Willd. var. latisecta D. F. Chamberlain. Their structures were elucidated by spectral methods. PMID- 12895548 TI - Comparative studies of the polysaccharides from species of the genus Ramalina lichenized fungi-of three distinct habitats. AB - Several structurally different glucans (alpha- and beta-) and galactomannans were characterized as components of four species of the genus Ramalina, namely R. dendriscoides, R. fraxinea, R. gracilis and R. peruviana. Freeze-thawing treatment of hot aqueous extracts furnished as precipitates (PW) linear alpha-D glucans of the nigeran type, with regularly distributed (1-->3)- and (1-->4) linkages in a 1:1 ratio. The supernatants (SW) contained alpha-D-glucans with (1- >3)- and (1-->4)-linkages in a molar ratio of 3:1. The lichen residues were then extracted with 2% aq. KOH, and the resulting extracts submitted to the freeze thawing treatment, giving rise to precipitates (PK2) of a mixture of alpha-glucan (nigeran) and beta-glucan, which were suspended in aqueous 0.5% NaOH at 50 degrees C, dissolving preferentially the beta-glucan. These were linear with (1- >3)-linkages (laminaran). The mother liquor of the KOH extractions (2% and 10% aq. KOH) was treated with Fehling's solution to give precipitates (galactomannans). The galactomannans are related, having (1-->6)-linked alpha-D mannopyranosyl main chains, substituted at O-4 and in a small proportion at O-2,4 by beta-D-galactopyranosyl units. Despite the different habitats of these lichenized fungi, all species studied in this investigation have a similar pool of polysaccharides. PMID- 12895549 TI - Neo-clerodane diterpenoids and phenylethanoid glycosides from Teucrium chamaedrys L. AB - A neo-clerodane type diterpenoid, 12(S)-15,16-epoxy-19-hydroxy-neo-cleroda 13(16),14-dien-18,6alpha:20,12-diolide, and two phenylethanoid glycosides, teucrioside-3(IIII)-O-methylether and teucrioside-3(IIII),4(IIII)-O-dimethylether were isolated from the aerial parts of Teucrium chamaedrys. Their structures were identified on the basis of extensive NMR spectra, LC-ESIMS analysis, and molecular modeling studies. PMID- 12895550 TI - Lignan and megastigmane glycosides from Sauropus androgynus. AB - A lignan diglycoside, (-)-isolariciresinol 3alpha-O-beta-apiofuranosyl-(1-->2)-O beta-glucopyranoside, and a megastigmane glucoside, sauroposide, were isolated from the aerial part of Sauropus androgynus together with (+)-isolariciresinol 3alpha-O-beta-glucopyranoside, (-)-isolariciresinol 3alpha-O-beta glucopyranoside, (+)-syringaresinol di-O-beta-glucopyranoside, guanosine and corchoionoside C. The structural elucidations were bases on analyses of physical and spectroscopic data. PMID- 12895552 TI - Partial characterization of the polluting load of swine wastewater treated with an integrated biodigestion system. AB - The stabilization of swine wastewaters from swine confined housing by the combination of a upflow anaerobic sludge blanket (UASB) reactor and waste stabilization ponds is a viable alternative to minimize the environmental impact caused by inadequate disposal of swine wastewaters. In the present study, the polluting load of pre-decanted swine wastewater treated with a series of two 0.705 m(3) UASB reactors and then in parallel in aerated and non-aerated stabilization tanks was investigated from January to July, 2000. Physicochemical and microbiological analyses were made adopting standard methods (Standard Methods for Examination of Water and Wastewater, 19th ed., American Public Health Association, Washington, DC, 1995). COD values decreased as the wastewater ran through the integrated biodigestion system dropping from about 3492+/-511-4094 mgl(-1)+/-481 to 124+/-52-490 mgl(-1)+/-230, while nitrate and nitrite levels increased in stabilization tanks, ranging respectively from 4+/-0 to 20 mgl(-1)+/ 3 and 3+/-1 to 11 mgl(-1)+/-24. Although the removal of Escherichia coli was more than 97%+/-6, the effluents of the treatment system still contained unacceptable levels of E. coli (1.6 x 10(3)-1.2 x 10(6) 100 ml(-1)) according to WHO guidelines for use of wastewater in agriculture and aquaculture. These results indicate the necessity of changes on operational characteristics of the treatment system such as an increase of the hydraulic retention time in UASB reactors or in stabilization tanks. PMID- 12895553 TI - Growth, biomass, carbon storage and nutrient distribution in Gmelina arborea Roxb. stands on red lateritic soils in central India. AB - Growth, biomass, carbon storage and nutrient (N, P and K) variations in 1 to 6 year-old chronosequence plantations of Gmelina arborea were studied in three degraded red lateritic sites in central India. Growth parameters (dbh, total height and number of branches) varied significantly due to difference in age and site quality, but tree density showed non-significant variation. Stand biomass ranged from 3.94 (1-year-old) to 53.67 Mgha(-1) (6-year-old) and stand carbon in 6-year-old plantations ranged from 24.12 to 31.12 Mgha(-1) at different sites. Among the tree components, the stem wood accounted for maximum C (56.25% at site 1) followed by branches (19.8% at site 3), roots (18.51% at site 2) and foliage (7.01% at site 3). Mean annual C accretion at 6 years age of plantation was highest in site 3 and it was 0.35, 2.66, 0.965 and 0.87 Mgha(-1) for leaf, stem, branches and roots, respectively. Quantity of nutrients increased with age. Total nitrogen accumulation in 6-year-old stands at the three sites ranged from 212.9 to 279.5 kgha(-1) with a mean annual storage of 238.43 kgha(-1) and total K ranged from 170.8 to 220.5 kgha(-1) with a mean annual storage of 189.93 kgha( 1). Phosphorous accumulation was lowest with a mean storage of 16.75 kgha(-1). The organic carbon and nutrients in the soils improved significantly after 6 years of G. arborea planting. Soil organic carbon increased from 8.46 to 14.02 Mgha(-1) within 6 years. At soil depths 0-20 cm, 21-40 cm and 41-60 cm, available N enhanced by 14.85%, 11.98% and 11.25%, K by 10%, 9.13% and 10.63%, whereas phosphorous declined by 26%, 23% and 20%, respectively. At 6 years, G. arborea stands sequestered 31.37 Mgha(-1) carbon. The nutrient management strategies in relation to carbon accretion in G. arborea stands on degraded lateritic sites are discussed. PMID- 12895554 TI - Mature green waste compost enhances growth and nitrogen uptake in wheat (Triticum aestivum L.) and oilseed rape (Brassica napus L.) through the action of water extractable factors. AB - A series of field and pot trials were carried out to determine the effects of growing wheat and oilseed rape in soils supplemented with green waste composts and provided with additional fertilisers. It was shown consistently that the response of wheat and rape to compost and fertiliser applied together was greater than the responses to the individual additives, but only when very stable compost was used (>10 months processing). Experiments with 15N-labelled fertiliser showed that wheat was able to utilise the applied N more efficiently when cultivated in the stable compost. The enhanced growth was also demonstrated in hydroponic culture of oilseed rape with water extracts of green waste compost in the presence of compound fertiliser. However the effect was rapidly lost at higher dilutions of compost extract (>3). It was concluded that water-extractable growth promoters are present in stable green waste compost, but these only have measurable activity at high concentrations. The identity of the growth promoting factors remains to be found, but the literature suggests that water-extractable humic substances or cytokinins may be involved. PMID- 12895555 TI - An examination of the effects of detergents on anaerobic digestion. AB - An anaerobic filter was used to examine the treatability of wastewater formulated to simulate that from the manufacture of detergents. The detergent element was a mixture (1:1:1 v/v) of concentrated washing up liquid, a non-biological hand washing detergent and a fabric softener and, thus contained a combination of cationic surfactants, anionic surfactants, non-ionic surfactants and amphoteric surfactants. A concentration of 2 ml/l caused a deterioration in the performance of the digester which was pronounced after 7 days. When the packing of the filter was modified to include two layers of granular activated carbon, 320 g in total, the reactor was capable of treating a feed containing 10 ml/l. The amount of detergent removed during this second trial was greater than the Langmuir monolayer capacity of the carbon, indicating that both adsorption and degradation were occurring. The results also showed that, at detergent concentrations greater than 1 ml/l, the theoretical COD removal was lower than the actual COD removal. This was compatible with there being an element of adsorption by the activated carbon. PMID- 12895556 TI - Synergetic effect of gamma irradiation and moisture content on decontamination of sewage sludge. AB - Samples of concentrated municipal sewage sludge, stored for 2, 4 and 6 months, with moisture contents of 2%, 20%, 40%, 60%, and 80% were exposed to doses of 0, 1, 2, 3, 4 and 5 kilogray (kGy) of gamma irradiation. Immediately after irradiation, total microbial count and bacterial pathogens in sewage sludge were determined. The results indicated that in all tested sewage sludge samples, bacterial pathogens including Enterobacter sp., Klebsiella sp., Salmonella sp., and Escherichia coli were initially detected. All doses of gamma irradiation reduced the total counts of microorganisms. D(10) of total count decreased with increase in the moisture content of the sewage sludge. The lowest lethal dose for tested bacterial pathogens was 5 kGy in air dried sewage sludge. In addition for wet sewage sludge having more than 40% moisture, the lethal dose was 1 kGy, for samples taken at different storage periods 2, 4 and 6 months, and therefore the cost per unit could be decreased to half when wet sewage sludge (about 50% moisture) was used. PMID- 12895557 TI - A simple procedure for preparing substrate for Pleurotus ostreatus cultivation. AB - The use of wooden crates for composting a mixture of 70% grass, (Digitaria decumbens), and 30% coffee pulp, combined with 2% Ca(OH)(2), was studied as a method for preparing substrate for the cultivation of Pleurotus ostreatus. Crate composting considerably modified the temperature pattern of the substrate in process, as compared to pile composting, where lower temperatures and less homogeneous distributions were observed. Biological efficiencies varied between 59.79% and 93% in the two harvests. Based on statistical analysis significant differences were observed between the treatments, composting times and in the interactions between these two factors. We concluded that it is possible to produce P. ostreatus on a lignocellulosic, non-composted, non-pasteurized substrate with an initial pH of 8.7, and that composting for two to three days improves the biological efficiency. PMID- 12895558 TI - Solids, organic load and nutrient concentration reductions in swine waste slurry using a polyacrylamide (PAM)-aided solids flocculation treatment. AB - Increased swine production results in concentration of wastes generated within a limited geographical area, which may lead to land application rates exceeding the local or regional assimilatory capacity. This may result in pollutant transfer through surface water or soil-groundwater systems, environmental degradation, and/or odor concerns. Existing swine waste pit storage and lagoon treatment technologies may be inadequate to store or treat waste prior to land application without these concerns resulting. Efficient swine waste solids separation may reduce environmental health concerns and generate a value-added bioresource (solids). This study evaluated the efficiency of a polyacrylamide (PAM) flocculant-aided solids separation treatment to reduce pollution indicator concentrations in raw (untreated) swine waste slurry. Swine waste slurry solids separation efficiency through gravity settling (sedimentation) was evaluated before and after the addition of a proprietary polymeric (PAM) flocculant. Results indicated that polymer amendments at concentrations of 62.5-750 mg/l improved slurry solids separation efficiency and significantly reduced concentrations of other associated aquatic pollution indicators in a majority of analyses conducted (33 of 50 total analyses conducted). Results also suggested that PAM-aided solids separation from swine waste slurry might facilitate further treatment and/or disposal and therefore reduce associated environmental degradation potential. PMID- 12895559 TI - Enhanced bioremediation of n-alkane in petroleum sludge using bacterial consortium amended with rhamnolipid and micronutrients. AB - The purpose of the present study was to investigate possible methods to enhance the rate of biodegradation of oil sludge from crude oil tank bottom, thus reducing the time usually required for bioremediation. Enhancement of biodegradation was achieved through bioaugmentation and biostimulation. About 10% and 20% sludge contaminated sterile and non-sterile soil samples were treated with bacterial consortium (BC), rhamnolipid biosurfactant (RL) and nitrogen, phosphorus and potassium (NPK) solution. Maximum n-alkane degradation occurred in the 10% sludge contaminated soil samples. The effects of treatment carried out with the non-sterile soil samples were more pronounced than in the sterile soils. Maximum degradation was achieved after the 56th day of treatment. n-Alkanes in the range of nC8-nC11 were degraded completely followed by nC12-nC21, nC22-nC31 and nC32-nC40 with percentage degradations of 100%, 83-98%, 80-85% and 57-73% respectively. Statistical analysis using analysis of variance and Duncan's multiple range test revealed that the level of amendments, incubation time and combination of amendments significantly influenced bacterial growth, protein concentration and surface tension at a 1% probability level. All tested additives BC, NPK and RL had significant positive effects on the bioremediation of n-alkane in petroleum sludge. PMID- 12895560 TI - Municipal solid waste management through vermicomposting employing exotic and local species of earthworms. AB - A comparative study was conducted between exotic and local (epigeic--Eisenia fetida and anaecic--Lempito mauritii, respectively) species of earthworms for the evaluation of their efficacy in vermicomposting of municipal solid waste (MSW). Vermicomposting of MSW for 42 days resulted in significant difference between the two species in their performance measured as loss in total organic carbon, carbon nitrogen ratio (C:N) and increase in total Kjeldahl nitrogen, electrical conductivity and total potassium and weight loss of MSW. The change in pH and increase in number of earthworms and cocoons and weight of earthworms were non significant. PMID- 12895561 TI - Adsorption of volatile organic compounds by pecan shell- and almond shell-based granular activated carbons. AB - The objective of this research was to determine the effectiveness of using pecan and almond shell-based granular activated carbons (GACs) in the adsorption of volatile organic compounds (VOCs) of health concern and known toxic compounds (such as bromo-dichloromethane, benzene, carbon tetrachloride, 1,1,1 trichloromethane, chloroform, and 1,1-dichloromethane) compared to the adsorption efficiency of commercially used carbons (such as Filtrasorb 200, Calgon GRC-20, and Waterlinks 206C AW) in simulated test medium. The pecan shell-based GACs were activated using steam, carbon dioxide or phosphoric acid. An almond shell-based GAC was activated with phosphoric acid. Our results indicated that steam- or carbon dioxide-activated pecan shell carbons were superior in total VOC adsorption to phosphoric acid-activated pecan shell or almond shell carbons, inferring that the method of activation selected for the preparation of activated carbons affected the adsorption of VOCs and hence are factors to be considered in any adsorption process. The steam-activated, pecan shell carbon adsorbed more total VOCs than the other experimental carbons and had an adsorption profile similar to the two coconut shell-based commercial carbons, but had greater adsorption than the coal-based commercial carbon. All the carbons studied adsorbed benzene more effectively than the other organics. Pecan shell, steam activated and acid-activated GACs showed higher adsorption of 1,1,1 trichloroethane than the other carbons studied. Multivariate analysis was conducted to group experimental carbons and commercial carbons based on their physical, chemical, and adsorptive properties. The results of the analysis conclude that steam-activated and acid-activated pecan shell carbons clustered together with coal-based and coconut shell-based commercial carbons, thus inferring that these experimental carbons could potentially be used as alternative sources for VOC adsorption in an aqueous environment. PMID- 12895562 TI - Sewage treatment by a low energy membrane bioreactor. AB - A new membrane bioreactor (MBR) was developed for treatment of municipal wastewater. The MBR was mainly made up of an activated sludge reactor and a transverse flow membrane module, with an innovative configuration being in application between them. As a result, the transverse flow membrane module and low recirculation flow rate created advantages, such as lower energy consumption and more resistance to membrane fouling. The total energy consumption in the whole system was tested as 1.97+/-0.74 kWh/m(3) (permeate) while using periodical backwash with treated water and backflush with mixed liquor daily, being in the same level as a submerged membrane bioreactor, reported to be 2.4 kWh/m(3) (permeate). Energy consumption analysis in the system shows that the membrane module was more energy consuming than the other four parts listed as pump, aeration, pipe system and return sludge velocity lose, which consumed 37.66 52.20% of the total energy. The effluent from this system could be considered as qualified for greywater reuse in China, showing its potential application in the future. PMID- 12895563 TI - Studies on adsorption of dyes on beta-cyclodextrin polymer. AB - Beta-cyclodextrin (beta-CD) polymers are used for the removal of various dyes from aqueous solutions. Three insoluble polymers with different degrees of beta CD were used. Results of adsorption experiments showed that these polymers exhibited high sorption capacities toward dyes. The mechanism of adsorption was both physical adsorption and hydrogen bonding due to the polymer and the formation of an inclusion complex due to the beta-CD molecules through host-guest interactions. PMID- 12895564 TI - Effect of feeding strategy on a stirred anaerobic sequencing fed-batch reactor containing immobilized biomass. AB - The present work reports on the influence of feeding strategy on the stability and performance of a stirred anaerobic sequencing fed-batch reactor containing biomass immobilized on polyurethane foam. The reactor treated low-strength wastewater and was operated at 30 degrees C with an agitation rate of 200 rpm. A 180-min cycle was used to treat approximately 0.5 l of synthetic substrate with a chemical oxygen demand concentration of nearly 500 mg/l. The reactor was operated in batch mode with a 3-min feeding step and in constant rate fed-batch mode with feeding steps of 30, 60 and 180 min. During batch operation, the system attained stability and had a removal efficiency of 86% based on non-filtered substrate concentration. However, during fed-batch operation stability and efficiency were impaired and formation of suspended material was identified. Stability was achieved only for the 30-min feeding step. The poor performance and instability observed in the fed-batch experiments were credited to the formation of considerable quantities of extracellular polymers, which impeded contact between substrate and biomass with consequent negative effect on the mass transfer fluxes. The biopolymer formation was very likely a result of the fed-batch operational mode, in which part of the bioparticles were deprived of contact with the liquid medium for a relatively long period of time. PMID- 12895565 TI - Highly thermostable amylase and pullulanase of the extreme thermophilic eubacterium Rhodothermus marinus: production and partial characterization. AB - Five strains of the extreme thermophilic Rhodothermus marinus were screened for the production of amylolytic and pullulytic activities. The culture medium for the selected strain, R. marinus ITI 990, was optimized using central composite designs for enhanced enzyme production. The optimized medium containing 1.5 gl( 1) of maltose and 8.3 gl(-1) of yeast extract yielded amylase, pullulanase and alpha-glucosidase activities of 45, 33 and 2.1 nkatml(-1), respectively. Among the various carbon sources tested, maltose was most effective for the formation of these enzymes, followed by soluble maize starch, glycogen and pullulan. The crude amylase and pullulanase showed maximum activities at pH 6.5-7.0, and 85 and 80 degrees C, respectively. At 85 degrees C amylase and pullulanase had half lives of 3 h and 30 min, respectively. PMID- 12895566 TI - Determination of some rheological parameters for the characterization of activated sludge. AB - The rheological characterization of activated sludge originating from different aeration tanks and from a lab-scale plant was carried out using rotation tests. Two rheological parameters were used: limit viscosity (mu(infinity)) and reduced hysteresis area (rHa). Mu(infinity) was strongly influenced by the total suspended solids (TSS) content of sludge and an exponential relation was found between TSS and the rheological parameter mu(infinity). Significant differences in sludge viscosity were found for sludge of different origins, but all of 10 g/l TSS content, indicating ability of viscosity to show different sludge qualities. Marked changes in activated sludge microorganisms communities were shown to have an influence on rheological parameters. During aerobic digestion of sludge, variations in mu(infinity) were noticed, indicating change of sludge quality. An over-growth of filamentous bacteria species was shown to induce a particular rheological behavior which could be characterized by rHa. This parameter was proposed as an index of filamentous bacteria proliferation in activated sludge aeration tanks. PMID- 12895567 TI - Replacement of maize by rumen filtrate fermented corn-cob in layer diets. AB - Laying hens (50 weeks in-lay) were fed for 8 weeks on diets containing corn-cobs fermented with rumen filtrate. The fermented corn-cob was included in the diets at 0%, 5.0%, 10.0%, 15.0% and 20.0% of the total diet at the expense of maize. Feed intake was not significantly (P>0.05) different, although birds on the control diet consumed slightly more feed than birds on the fermented corn-cob based diets. Hen day production and feed per dozen eggs decreased with increase in dietary level of corn-cob (P<0.05). Yolk colour was significantly (P<0.05) affected, rumen filtrate fermented corn-cob appear to promote better yolk coloration. Egg weight and shell thickness were not significantly influenced by dietary treatment. The overall results appear to suggest that rumen filtrate fermentated corn-cob could provide a material that holds promise as a good alternative to maize because of its enhanced crude protein value and reduced crude fibre content. PMID- 12895568 TI - Influence of precipitated calcium carbonate (CaCO3) on shiitake (Lentinula edodes) yield and mushroom size. AB - Synthetic substrate consisting of oak sawdust (50%), white millet (28%), winter rye (11%) and soft red wheat bran (11%) was non-supplemented or supplemented with 0.2%, 0.4% or 0.6% (dry weight basis) precipitated calcium carbonate (CaCO(3)). Shiitake (Lentinula edodes) was grown in two crops to determine the effect of three CaCO(3) levels on mushroom yield and size. Yields and biological efficiencies (averages for two crops) from substrates non-supplemented with CaCO(3) were lower by 14.1%, 18.4% and 24.9% compared to treatments supplemented with 0.2%, 0.4% and 0.6% CaCO(3), respectively. Mushroom size (weight) was larger with non-supplemented substrate (16.8 g) compared to substrate supplemented with 0.6% CaCO(3) (15.1 g). However, mushroom production was more consistent from crop to crop when 0.6% CaCO(3) was added to substrate. PMID- 12895569 TI - Osteoblastic cell response to thin film of poorly crystalline calcium phosphate apatite formed at low temperatures. AB - The response of osteoblastic cells to a thin film of poorly crystalline calcium phosphate apatite crystals (PCA) was examined in vitro. The PCA thin film was prepared on polystyrene culture dishes using highly metastable calcium phosphate ion solution at low temperatures. The PCA thin film was formed through fusion and transformation of granular calcium phosphate particles, which had initially formed on the surface, into a film of calcium phosphate apatite crystal. The PCA thin film was used for cell culture without additional surface treatment. The osteoblastic cell behaviors including adhesion, proliferation, expression of the marker genes, and calcified matrix formation were examined on the PCA thin film using primary osteoblasts or MC3T3-E1 cells. The cells were well attached and had spread in a slender shape over the PCA thin film. The extent of cell proliferation on the PCA thin film is as much as on the plain dishes. In addition, a much larger number of calcified nodules had formed on the PCA thin film than on the plain dish. The expression of the marker genes such as alkaline phosphatase, osteocalcin, osteopontin, osteonectin was apparent. These results demonstrate that the osteoblasts exhibit a full spectrum of cellular activity including the adequate differentiation on the PCA thin film. Therefore, a PCA thin film can be used as a coating material for biomaterials where the surface is not adequate for inducing the full activity of bone cells. PMID- 12895570 TI - Synthesis, characterization, and blood compatibility of polyamidoamines copolymers. AB - This work reports the development of new non-thrombogenic polymers based on the linear polymers of polyamidoamines (PAAs), having heparin binding ability, obtained by polyaddition of secondary amines to N,N'-methylene bis-acrylamide. PAAs could not be used directly in the making of blood-contacting materials due to their poor mechanical strength. In order to overcome this lacuna, copolymers of amidoamine with methylmethacrylate (MMA) were prepared. Characterization studies indicated that the PAAs have been suitably incorporated into the MMA matrix. The relative hydrophilic nature of the synthesized copolymers was established from the measurement of water contact angle. The heparinized copolymers showed significant improvement in non-thrombogenic characteristics. PMID- 12895571 TI - The exothermal behavior in the hydration process of calcium phosphate cement. AB - The exothermal behavior in the hydration process of calcium phosphate cement (CPC) and their effect factors were investigated systematically. One CPC formulation is composed of tetracalcium phosphate (TECP) and dicalcium phosphate anhydrous (DCPA), and its hydration reaction is an exothermic reaction. Hydration includes five processes, namely, the start, induction, acceleration, deceleration and the terminating process. The introduction of hydroxyapatite (HAP) as crystal seed shortens induction and advances on the reaction peak position, which resulted from the introduction of crystal seed that reduced the activation energy of heterogeneous nucleation. The increase in temperature not only makes the reaction advance but also enlarges it. The activation energy of the hydration reaction is 176kJ/mol by calculation. The particle size of the raw material and the ratio of calcium to phosphate in the specimen have large effects on the hydration process. Decrease in particle size of TECP and in the ratio of calcium to phosphate resulted in marked acceleration of reaction and made the reaction peak advance evidently, but the effect of particle size of DCPA is not so remarkable. Based on the heat liberation in the hydration of CPC specimens taking Ca/P as 1.5 and 1.67, the standard mole enthalpies of formation of TECP and calcium-deficient hydroxyapatite are -5908 and -11,119kJmol(-1), respectively. PMID- 12895572 TI - Synergistic effect of bismuth subgallate and borneol, the major components of Sulbogin, on the healing of skin wound. AB - Most skin lesions heal delay and even heal efficiently within 1-2 weeks, the healed tissue is neither aesthetically nor functionally perfect. Therefore, facilitating skin healing rate and controlling healed skin quality are major aims of drug treatment for a wound event. Bismuth subgallate (BS) and Borneol (BO) are the two components of Sulbogin, a new Vaseline-based wound healing ointment, one for treating skin wounds. Although BO has antibiotic function, while BS is widely used clinically, neither has been used specifically for wound healing. The experiment described here aimed to study the effect of BS and BO on the healing of skin wounds. This study also compared the effects of BS and BO with Flamazine cream, which is currently the most popular drug for wound healing in hospitals. Full-thickness wounds (3 cm x 3 cm x 0.2 cm) were created on the back of adult male Sprague-Dawley rats. BS, BO, BS+BO, and Flamazine were then evenly applied to cheesecloth and placed over the lesion areas. The drug patches were replaced every 2-3 days until the wound areas were completely covered by epidermis in any kinds of drug treatment. The combined BS and BO treatment had the best effect on healing by decreasing lesion area, while increasing granulation tissue formation, re-epithelialization, eating behavior and reconstitution of skin appendages. This investigation showed that BS and BO have a synergistic effect on the skin wound restoration. PMID- 12895573 TI - Staphylococcus epidermidis-fibronectin binding and its inhibition by heparin. AB - Staphylococcus epidermidis is able to adhere onto biomaterials and to cause implant infections. Recently, host matrix proteins, which in vivo cover the implants, have been indicated as substrates for adhesion by specific bacterial adhesins. Here, the binding of S. epidermidis to fibronectin, a main protein of the extracellular matrix, and the effect of heparin on this interaction were studied by dynamic force spectroscopy (DFS). Novelties are that S. epidermidis strains analysed by DFS were clinical isolates from prosthesis-associated infections, genotyped and phenotyped for their adhesion properties to fibronectin and examined as living cells. Thus, fibronectin-binding staphylococci adhered to the fibronectin-coated substratum and formed a continuous layer assuring their contact with the fibronectin-coated cantilever tip during the approach-retraction cycles of the DFS measurements. Results show that only a single molecular binding site of fibronectin is involved in the interaction with S. epidermidis, that it takes place at the domain near the C-terminus and that it is specifically inhibited by heparin. PMID- 12895574 TI - Preparation of a protein micro-array using a photo-reactive polymer for a cell adhesion assay. AB - A protein micro-array, called a "cell chip" was constructed by using a photo reactive polymer for a cell-adhesion assay. Various amounts of albumin or fibronectin were covalently immobilized on a polystyrene dish using a micro spotter with a dip pen. First, poly(acrylic acid) carrying azidophenyl groups was synthesized as the photo-reactive polymer. Secondly, the aqueous solution of a photo-reactive polymer (several nanoliters) was cast using the dip pen of the micro-spotter and dried in air. Subsequently, aqueous solutions of proteins were cast on the same place using the micro-spotter. After drying, the dish was irradiated with ultraviolet light. Finally, the immobilization was confirmed by staining with a dye. The immobilization was stable even after washing with Tween 20. The protein-immobilized area depended on the manipulation of the micro spotter and the size of the dip pen. Subsequently, cell adhesion on the photo immobilized protein micro-array was investigated. The adhesion behavior of cells depended on the kind of immobilized proteins and the kind of cells. The protein micro-array will be useful for cell diagnosis and for the selection of biomaterials to regulate cell behavior. PMID- 12895575 TI - Evaluation of corrosion on plasma sprayed and anodized titanium implants, both with and without bone cement. AB - The corrosion behavior of titanium with vacuum plasma sprayed titanium coatings and with anodized surfaces, both with and without polymeric bone cement were evaluated. Electrochemical extraction tests were carried out with subsequent analysis of the electrolyte by ICP-MS in order to verify our hypothesis of the ionic permeability of the polymer cement. The complexity of the situation resides in the existence of two interfaces: electrolyte-polymer and polymer-metal. The surface preparation (treatment of the surface) plays an important role in the corrosion resistance of titanium. The electrochemical magnitudes that were examined reveal that the plasma spray surfaces have the lowest corrosion resistance. The cement, in spite of having reduced electrical conductivity in comparison to metal, is an ionic transporter, and therefore capable of participating in the corrosion process. In the present study, we observed in fact crevice corrosion at the metal-cement interface. In the case of plasma spray surfaces, a process of diffusion of titanium particles in the electrolyte could accompany the crevice corrosion. In this study, we have shown that there is a corrosion process at the surface of the titanium through the cement which has as a consequence on the one hand the formation of titanium cations and on the other hand the growth of a passive layer on the titanium. In conclusion, we identified two principal factors that influence the corrosion process: [1] the type of surface treatment for the titanium, and [2] the ionic conductivity of the cement. There is indeed ionic transport through the cement; as evidenced by the presence of titanium in the electrolyte solution (ICP-MS analysis) and chloride at the surface of the titanium sample (EDX analysis). We show that the polymer cement is an ionic conductor and participates in the corrosion of the embedded titanium. We cannot deduce from our results, however, whether the polymer itself possesses corrosive properties. Long-term experiments will be necessary to study the degradation behavior of the polymer cement. PMID- 12895576 TI - Initial in vitro interaction of osteoblasts with nano-porous alumina. AB - In the present study we have used a characterised primary human cell culture model to investigate cellular interactions with nano-porous alumina. This material, prepared by anodisation, is being developed as a coating on titanium alloy implants. The structure of the alumina, as determined by X-ray diffraction and transmission electron microscopy, was amorphous. When studying cell/material interactions we used both biochemical and morphological parameters. Cell viability, proliferation and phenotype were assessed by measurement of redox reactions in the cells, cellular DNA, tritiated thymidine ([3H]-TdR) incorporation and alkaline phosphatase (ALP) production. Results showed a normal osteoblastic growth pattern with increasing cell numbers during the first 2 weeks. A peak in cell proliferation was seen on day 3, after which cell growth decreased, followed by an increase in ALP production, thus indicating that the osteoblastic phenotype was retained on the alumina. Cell adhesion was observed, the osteoblast-like cells having a flattened morphology with filipodia attached to the pores of the material. SDS-PAGE and western blot measurements showed that the nano-porous alumina was able to adsorb fibronectin. Trace amounts of aluminium ions were measured in the surrounding medium, but no adverse effect on cell activity was observed. PMID- 12895577 TI - The effect of frictional heating and forced cooling on the serum lubricant and wear of UHMW polyethylene cups against cobalt-chromium and zirconia balls. AB - Hip simulator tests of femoral balls of cobalt-chromium alloy or zirconia against acetabular cups of UHMW polyethylene were run with and without a coolant circulated inside the femoral balls. Without cooling, the wear of polyethylene against zirconia was about 48% lower than with cobalt-chromium alloy, but the steady-state temperature of the zirconia ball was higher (55 degrees C vs. 41 degrees C), and there was more precipitation of protein from the serum, which sometimes formed an adherent layer on the surface of the zirconia. Circulating coolant at 1-20 degrees C markedly reduced the bearing temperatures and the protein precipitation. With coolant at 4 degrees C, wear of the polyethylene against cobalt-chromium alloy was about 26% lower than against zirconia, but the macroscopic and microscopic appearance of the worn polyethylene surfaces were unlike that typically generated in vivo. With or without coolant, the morphology of the polyethylene wear debris was comparable to that generated in vivo, but the ratio of fibrillar to granular debris was higher at the reduced temperature. These results suggested that circulating coolant at an appropriate temperature could avoid overheating (due to non-stop running of the simulator), preventing excessive protein precipitation while providing wear surfaces and wear debris with morphologies closely comparable to those generated in vivo. PMID- 12895578 TI - Effects of added bioactive glass on the setting and mechanical properties of resin-modified glass ionomer cement. AB - In this study, the effects of added bioactive glass on the basic setting properties of a commercially available resin-modified glass ionomer cement were investigated with respect to setting time, mechanical strength, and setting mechanism. It was found to be clinically acceptable whether the setting time was extended or shortened depending on the type of bioactive glass added. The compressive strength of the set cement containing the bioactive glass decreased and was much higher when compared with the conventional type glass ionomer cement containing bioactive glass. The Fourier-transform infrared and 13C CP/MAS-NMR spectroscopies revealed that the extent of the acid-base reaction was larger in the cements containing bioactive glass than in the commercial resin-modified glass ionomer cement because of its high basicity in the bioactive glass. The 27Al MAS-NMR showed that crosslinking of the carboxylates in the polymeric acid by Al proceeded less in the cement containing the bioactive glass. PMID- 12895579 TI - Nucleation and growth of apatite on chemically treated titanium alloy: an electrochemical impedance spectroscopy study. AB - Bone-like apatite formed on the surface of Ti6Al4V pretreated with NaOH solution after having been immersed in simulated body fluid (SBF), while no apatite formed on the surface of untreated Ti6Al4V. In the present study, electrochemical impedance spectroscopy (EIS) measurement was used to investigate the nucleation and growth of apatite on chemically treated Ti6Al4V immersed in the SBF solution, and the difference between the behaviors of treated and untreated Ti6Al4V. Appropriate equivalent circuit models were constructed to describe the nucleation and growth of apatite, and thin oxide film formed on the surface of untreated Ti6Al4V. It was found that EIS is a useful method for investigating the nucleation and growth of bone-like apatite on Ti6Al4V pretreated with NaOH solution. PMID- 12895580 TI - Macrophage responses to silk. AB - Silk fibers have potential biomedical applications beyond their traditional use as sutures. The physical properties of silk fibers and films make it a promising candidate for tissue engineering scaffold applications, particularly where high mechanical loads or tensile forces are applied or in cases where low rates of degradation are desirable. A critical issue for biomaterial scaffolds is biocompatibility. The direct inflammatory potential of intact silk fibers as well as extracts was studied in an in vitro system. The results indicate that silk fibers are largely immunologically inert in short- and long-term culture with RAW 264.7 murine macrophage cells while insoluble fibroin particles induced significant TNF release. Soluble sericin proteins extracted from native silk fibers did not induce significant macrophage activation. While sericin did not activate macrophages by itself, it demonstrated a synergistic effect with bacterial lipopolysaccharide. The low level of inflammatory potential of silk fibers makes them promising candidates in future biomedical applications. PMID- 12895581 TI - Conformational change of hydroxyapatite/gelatin nanocomposite by glutaraldehyde. AB - The hydroxyapatite (HAp)/gelatin (GEL) nanocomposite was prepared through the coprecipitation and then cross-linked by using glutaraldehyde (GA). From FT-IR measurement the spectral features for amide bands and phosphate bands were severely modified by the cross-linkage and the organic content increased with the degree of cross-linkage. From Transmission Electron Microscopy (TEM) and electron diffraction analyses we could confirm the preferentially directional growth of needle-like HAp particles, which were embedded in GEL by the mineralization. Also we could observe worm-like stria patterns of contrast, which were revealed by the mineralization on the individual GEL fibrils. Moire images were observed in a highly cross-linked HAp/GEL nanocomposite sample and we think the cross-linkage induced the assembly of unordered individual fibrils along the preferential direction. PMID- 12895582 TI - Platelet-derived growth factors enhance proliferation of human stromal stem cells. AB - Studies on new procedures for bone reconstruction suggest that autologous cells seeded on a resorbable scaffold can improve the treatment of bone defects. It is important to develop culture conditions for ex vivo expansion of stromal stem cells (SSC) that do not compromise their self-renewing and differentiation capability. Bone marrow SSC and platelet gel (PG) obtained by platelet-rich plasma provide an invaluable source for autologous progenitor cells and growth factors for bone reconstruction. In this study the effect of platelet-rich plasma (PRP) released by PG on SSC proliferation and differentiation was investigated. MTT assay was used to investigate the effect of PRP on proliferation: results showed that PRP induced SSC proliferation. The effect was dose dependent and 10% PRP is sufficient to induce a marked cell proliferation. Untreated cells served as controls. Upon treatment with 10% PRP, cells entered logarithmic growth. Removal of PRP restored the characteristic proliferation rate. Because SSC can gradually lose their capability to differentiate along the chondrogenic and osteogenic lineage during subculture in vitro, we tested whether 10% PRP treatment affected SSC ability to mineralize. SSC were first exposed to 10% PRP for five passages, at passage 6 PRP was washed away and plated cells were treated with dexamethasone (DEX). DEX induced a three-fold increase in the number of alkaline phosphatase positive cells and induced mineralization that is consistent with the differentiation of osteochondroprogenitor cells. In conclusion, 10% PRP promotes SSC proliferation; cells expanded with 10% PRP can mineralize the extracellular matrix once PRP is withdrawn. PMID- 12895583 TI - Multifunctional microcapsules for pancreatic islet cell entrapment: design, preparation and in vitro characterization. AB - Great advances in cell transplantation have been made, including the recent, remarkable success in pancreatic islet transplantation for the treatment of type 1 diabetes mellitus. Unfortunately, the transplanted cells are very susceptible to oxidative stress that cause severe damage to either allo- or xenogeneic islets upon graft in diabetic patients. Consequently, the transplanted islet functional life span is significantly shortened. The aim of this study was to examine the possible effects of antioxidants on in vitro cultured adult rat islets, and to evaluate the effects of a prolonged-release formulation, in form of cellulose acetate (CA) microspheres, on Vitamin D(3) activity. Isolated rat islets, both free and entrapped in microspheres were treated with Vitamin D(3). The effects of the vitamin were studied at 3, 6 and 9 days of in vitro cell culture. According to insulin secretory patterns, treatment with Vitamin D(3) of both free and CA entrapped microspheres, increased the insulin output as compared to untreated controls. Such positive effects were confirmed under islet static incubation with glucose at day 6. These results suggest that pancreatic islets can be advantageously treated with anti-oxidising vitamins before implantation, and speculatively, with the help of special delivery systems, throughout the islet cell life span, in the post-transplant time period. PMID- 12895584 TI - Scaffold development using selective laser sintering of polyetheretherketone hydroxyapatite biocomposite blends. AB - In tissue engineering (TE), temporary three-dimensional scaffolds are essential to guide cell proliferation and to maintain native phenotypes in regenerating biologic tissues or organs. To create the scaffolds, rapid prototyping (RP) techniques are emerging as fabrication techniques of choice as they are capable of overcoming many of the limitations encountered with conventional manual-based fabrication processes. In this research, RP fabrication of solvent free porous polymeric and composite scaffolds was investigated. Biomaterials such as polyetheretherketone (PEEK) and hydroxyapatite (HA) were experimentally processed on a commercial selective laser sintering (SLS) RP system. The SLS technique is highly advantageous as it provides good user control over the microstructures of created scaffolds by adjusting the SLS process parameters. Different weight percentage (wt%) compositions of physically mixed PEEK/HA powder blends were sintered to assess their suitability for SLS processing. Microstructural assessments of the scaffolds were conducted using electron microscopy. The results ascertained the potential of SLS-fabricated TE scaffolds. PMID- 12895585 TI - Tissue engineering of white adipose tissue using hyaluronic acid-based scaffolds. I: in vitro differentiation of human adipocyte precursor cells on scaffolds. AB - BACKGROUND AND AIM OF THE STUDY: Reconstruction of soft tissue defects is a challenge in plastic surgery and there is clinical need for adequate solutions. Aim of this study was to develop a biohybrid construct consisting of hyaluronic acid-based scaffolds and human adipocyte precursor cells as a soft tissue filler. METHODS: Human adipocyte precursor cells were obtained by collagenase digestion of adipose tissue samples and seeded on hyaluronic acid-based spongy scaffolds of various degrees of esterification and pore size using different techniques. After cell attachment, adipose differentiation was induced by defined adipogenic factors under serum-free culture conditions. RESULTS: Among the five different scaffold types under investigation the highest cell attachment rate was observed for the HYAFF scaffold with 100% esterification and a mean pore size of 400microm (HYAFF 11lp). For inoculation of human adipocyte precursor cells on hyaluronic acid-based scaffolds a "drop-on" technique and low-pressure centrifugation using a Speed Vac airfuge were compared. With respect to efficacy, cell distribution and simpleness the drop-on method proved to be the method of choice. In a serum free medium supplemented with 66nM insulin, 100nM cortisol and 1microg/ml troglitazone a substantial proportion of cells underwent adipose differentiation as assessed by lipid accumulation and emergence of glycerol-3-phosphate dehydrogenase activity, a lipogenic marker enzyme. CONCLUSION: Hyaluronic acid based scaffolds appear to be a suitable three-dimensional carrier for the culture and in vitro differentiation of human adipocyte precursor cells. PMID- 12895586 TI - In situ pore formation in a polymer matrix by differential polymer degradation. AB - A new approach for the in situ formation of porosity in a matrix based on differential polymer degradation has been studied. This approach exploits the differences in polymer properties such as molecular weight, hydrophilicity (hydrophobicity), and degradation to induce preferential degradation of one phase in a biphasic polymer system. Biphasic polymer systems polymers derived from alpha-hydroxy acids and poly(anhydrides), which vary in their erosion characteristics (surface vs. bulk) and hydrophobicities were studied. In addition to examining the generality of the approach, potential advantages of such systems in the context of tissue engineering and drug delivery are briefly discussed. PMID- 12895587 TI - Physiological strains induce differentiation in human osteoblasts cultured on orthopaedic biomaterial. AB - We have developed an in vitro mechanical stretching model of osteoblastic cells cultured on metallic biomaterials in order to study the effects of mechanical strain on osteointegration of orthopaedic implants. Titanium alloy discs coated with alumina or hydroxyapatite were used as substrates. Three Dynacell devices were especially designed to apply cyclic strains on rigid biomaterials. The regimen (600 mu epsilon strains, 0.25Hz) was defined on the basis of physiological data and estimated deformation on hip stem prostheses. The performances of these apparatus were reproducible and provided controlled deformations. Human osteosarcoma cell line MG-63, human osteoblasts obtained from primary cultures and ROS 17/2.8 rat osteosarcoma cells were used as cell models. Cell behaviour was assessed in terms of growth and alkaline phosphatase (ALP) activity by in situ assays for two regimens: 15-min deformations repeated three times a day to mimic rehabilitation exercises and 24-h continuous deformations. We demonstrated that continuous deformation did not affect the growth and ALP activity of MG-63 cells, in contrast with sequential deformations which had no effect on cell number, but which stimulated ALP activity after 5 days of stretching. This sequential regimen can also modify the behaviour of human bone derived cells resulting in increased proliferation after 5 days and stimulation of ALP activity after 15 days. ROS 17/2.8 rat osteosarcoma cells submitted to sequential deformations responded faster than other cell lines by increasing their ALP activity only after 1 day of stretching. Like MG-63 cells, proliferation of the ROS 17/2.8 rat osteosarcoma cell line was not affected by sequential deformations. This study suggests that short, repeated deformations defined to mimic rehabilitation exercises recommended after prostheses implantation are more likely to exert beneficial effects on implanted bone than continuous strains. PMID- 12895588 TI - Devitrification of ionomer glass and its effect on the in vitro biocompatibility of glass-ionomer cements. AB - The effects of devitrification of an ionomer glass with a molar composition 4.5SiO(2).3Al(2)O(3).1.5P(2)O(5).3CaO.2CaF(2) on cement formation and in vitro biocompatibility were investigated. Differential thermal analysis was used to study the phase evolution in the glass, and to determine the heat treatments for production of glass-ceramics. X-ray diffraction patterns from glass frit heat treated at 750 degrees C for 2h contained peaks corresponding to apatite (JCPDS 15-876), whereas for samples heat-treated at 950 degrees C for 2h apatite and mullite (JCPDS 15-776) were the major phases detected. Transmission electron microscopy (TEM) confirmed that apatite and apatite-mullite phases were present after heat treatments at 750 degrees C and 950 degrees C respectively. Glass and glass-ceramics were ground to prepare <45microm powders and glass ionomer cements were produced using a ratio of 1g powder: 0.2g PAA: 0.3g 10% m/v tartaric acid solution in water. In vitro biocompatibility was evaluated using cultured rat osteosarcoma (ROS) cells. Scanning electron microscopy (SEM) showed that cells colonised the surfaces of cements prepared using untreated ionomer glass and glass crystallised to form apatite (750 degrees C/2h). However, quantitative evaluation using MTT and total protein assays indicated that more cell growth occurred in the presence of cements prepared using ionomer glasses crystallised to apatite than cements prepared using untreated glass. The least cell growth and respiratory activity was observed on cements made with crystallised glass containing both apatite and mullite. It was concluded that the controlled devitrification of ionomer glasses could be used to produce GIC bone cements with improved biocompatibility. PMID- 12895589 TI - Mechanical evaluation of novel spinal interbody fusion cages made of bioactive, resorbable composites. AB - Osteoconductive and totally bioresorbable spinal/cervical interbody fusion cages were fabricated from a forged composite of raw particulate hydroxyapatite/poly L lactide (u-HA/PLLA) with an u-HA 40wt% fraction (F-u-HA 40). The mechanical strengths of three types of cages, designed for open-box, screw and cylinder constructs, were compared with those of existing metal and carbon-fiber/polymer cages. Compressive strengths of these composite cages surpassed those of existing metal and carbon-fiber cages. Fatigue resistance to alternate and static compressive loading persisted for longer than the minimum period (6 months) necessary for spinal devices in simulated body fluid (SBF) at 37 degrees C. These novel interbody fusion cages await clinical application in humans. PMID- 12895591 TI - Phospholipase A(2). AB - Considerable progress has been made in characterizing the individual participant enzymes and their relative contributions in the generation of eicosanoids, lipid mediators derived from arachidonic acid, such as prostaglandins and leukotrienes. However, the role of individual phospholipase (PL) A(2) enzymes in providing arachidonic acid to the downstream enzymes for eicosanoid generation in biologic processes has not been fully elucidated. In this review, we will provide an overview of the classification of the families of PLA(2) enzymes, their putative mechanisms of action, and their role(s) in eicosanoid generation and inflammation. PMID- 12895592 TI - 5-lipoxygenase and FLAP. AB - The initial steps in the biosynthesis of leukotrienes from arachidonic acid are carried out by the enzyme 5-lipoxygenase (5-LO). In intact cells, the helper protein 5-LO activating protein (FLAP) is necessary for efficient enzyme utilization of endogenous substrate. The last decade has witnessed remarkable progress in our understanding of these two proteins. Here we review the molecular and cellular aspects of the expression, function, and regulation of 5-LO and FLAP. PMID- 12895593 TI - Leukotriene C(4) synthase. AB - LTC(4) synthase conjugates LTA(4) with glutathione (GSH) to form LTC(4), the parent compound of the cysteinyl leukotrienes. LTC(4) synthase is a membrane protein that functions as a non-covalent homodimer of two 18-kDa polypeptides. The enzymatic activity of LTC(4) synthase is augmented by Mg(2+) and inhibited by Co(2+) and the FLAP inhibitor MK-886. The K(m) and V(max) values of human LTC(4) synthase are 3.6 microM and 1.3 micromol/mg/min for LTA(4) and 1.6 mM and 2.7 micromol/mg/min for GSH, respectively. The deduced amino acid sequence and the predicted secondary structure of LTC(4) synthase share significant homology to FLAP, mGST-2, and mGST-3. Site-directed mutagenesis of LTC(4) synthase suggests that Arg-51 is involved in opening the epoxide ring of LTA(4) and Tyr-93 in GSH thiolate anion formation during catalytic conjugation. LTC(4) synthase is a TATA less gene whose transcription involved both cell- and non-specific regulatory elements. LTC(4) synthase gene disrupted mice grow normally, and are attenuated for innate and adaptive immune inflammatory permeability responses. PMID- 12895594 TI - The cysteinyl leukotriene receptors. AB - The cysteinyl leukotriene (CysLT) receptors are putative 7 transmembrane spanning G protein-coupled receptors (GPCRs) of the rhodopsin subfamily of GPCRs. Two human and mouse CysLT receptors have been molecularly cloned and characterized. The properties of these receptors agrees well with previous pharmacological CysLT agonist and antagonist characterizations of the CysLT receptors. PMID- 12895595 TI - BLT1 and BLT2: the leukotriene B(4) receptors. AB - Two receptors for leukotriene B(4) (LTB(4)) have been molecularly identified: BLT1 and BLT2. Both receptors are G protein-coupled seven transmembrane domain receptors, whose genes are located in very close proximity to each other in the human and mouse genomes. The two receptors differ in their affinity and specificity for LTB(4): BLT1 is a high-affinity receptor specific for LTB(4), whereas BLT2 is a low-affinity receptor that also binds other eicosanoids. The two receptors also differ in their pattern of expression with BLT1 being expressed primarily in leukocytes, whereas BLT2 is expressed more ubiquitously. By mediating the activities of LTB(4), these receptors participate both in host immune responses and in the pathogenesis of inflammatory diseases. Reduced disease severity in animal inflammatory models seen with LTB(4) receptor antagonists and in mice with targeted deletion of BLT1 have revealed important roles for LTB(4) and its receptors in regulating pathologic inflammation. PMID- 12895596 TI - Eosinophils and cysteinyl leukotrienes. AB - Eosinophils are the main source of the cysteinyl leukotrienes, LTC(4)/D(4)/E(4), which are lipid mediators that play major roles in the pathogenesis of asthma and other forms of allergic inflammation. Here, we review the mechanisms governing eosinophil LTC(4) synthesis, focusing on the distinct intracellular domains that regulate eicosanoid formation and function within eosinophils. Cysteinyl leukotrienes exert their actions by engaging specific receptors. As recently shown, eosinophils express CysLT1 and CysLT2, the only cloned receptors for cysteinyl leukotrienes. Therefore, here we also present some of the new findings regarding the paracrine/autocrine activation of these CysLT receptors on eosinophils, and discuss some data on novel intracrine effects of LTC(4) triggered by a putative third CysLT receptor expressed intracellularly within eosinophils. PMID- 12895597 TI - The role of leukotrienes in asthma. AB - Leukotrienes (LT), both the cysteinyl LTs, LTC(4), LTD(4) and LTE(4), as well as LTB(4) have been implicated in the clinical course, physiologic changes, and pathogenesis of asthma. The cysteinyl LTs are potent bronchoconstrictors, which have additional effects on blood vessels, mucociliary clearance and eosinophilic inflammation. In addition, the cysteinyl LTs are formed from cells commonly associated with asthma, including eosinophils and mast cells. LTB(4), whose role is less well defined in asthma, is a potent chemoattractant (and cell activator) for both neutrophils and eosinophils. In the last 5 years, drugs have been developed which block the actions or formation of these mediators. Clinical and physiologic studies have demonstrated that they are modest short-acting bronchodilators, with sustained improvement in FEV(1) occurring in double-blind, placebo-controlled clinical trials for up to 6 months. These drugs have demonstrated efficacy in preventing bronchoconstriction caused by LTs, allergen, exercise and other agents. Additionally, there are multiple published studies which have demonstrated improvement in asthma symptoms, beta agonist use and, importantly, exacerbations of asthma in both adults and children. Comparison studies with inhaled corticosteroids (ICS) suggest that ICS are superior to leukotriene modifying drugs in moderate persistent asthma. However, several published studies now suggest that leukotriene modifying drugs are effective when added to ongoing therapy with ICS, either to improve current symptoms or to decrease the dose of ICS required to maintain control. While an anti-inflammatory effect is suggested, longer-term, earlier intervention, studies are needed to determine whether these compounds will have any effect on the natural history of the disease. PMID- 12895598 TI - Cyclooxygenase enzymes in allergic inflammation and asthma. AB - The cyclooxygenase enzyme system produces eicosanoids which mediate many important physiological and pathological functions. Experimental and clinical data suggest a role for this enzyme system in the pathogenesis of allergic inflammation and asthma. This article focuses on the function of this pathway in the lung, reviews evidence implicating the involvement of this pathway in asthma and allergic airway inflammation, and discusses implications for the treatment of asthmatics with cyclooxygenase inhibitors. PMID- 12895599 TI - Hematopoietic prostaglandin D synthase. AB - The biological actions of prostaglandin (PG) D(2) include vasodilatation, bronchoconstriction, inhibition of platelet aggregation, and recruitment of inflammatory cells. Characterization of DP receptor null mice in which antigen induced airway and inflammatory responses are attenuated and identification of CRTH2 as a novel PGD(2) receptor have shed light on the role of PGD(2) in the immune and inflammatory responses. Hematopoietic PGD synthase (H-PGDS) is a cytosolic enzyme that isomerizes PGH(2), a common precursor for all PGs and thromboxanes, to PGD(2) in a glutathione-dependent manner. H-PGDS is expressed in mast cells, antigen-presenting cells, and Th2 cells, and is the only mammalian member of the Sigma class of cytosolic glutathione S-transferases. In this review, we focus on the molecular biology of H-PGDS, the determination of its three-dimensional structure, characterization of the regulation of its gene expression, and information gleaned from transgenic animals. PMID- 12895600 TI - The second PGD(2) receptor CRTH2: structure, properties, and functions in leukocytes. AB - Prostaglandin (PG) D(2) plays a broad range of physiological and pathophysiological functions. Until just a few years ago, it was thought that most of the biological actions of PGD(2) are mediated via the classical PGD(2) receptor DP. Recently, we identified a second PGD(2) receptor, chemoattractant receptor-homologous molecule expressed on T helper (Th)2 cells (CRTH2), with different functions relative to DP. Here, we review the recent findings on the structure, tissue distribution, ligand selectivity, signalling pathways, and functions in leukocytes of this receptor. The data suggest that the PGD(2)/CRTH2 system play important roles in allergic inflammation through its stimulatory effects on Th2 cells, eosinophils, and basophils. PMID- 12895601 TI - A novel PGD(2) receptor expressed in eosinophils. AB - PGD(2) is a major product of arachidonic acid metabolism by mast cells and is released in the lungs following allergen challenge. Activation of the classic PGD(2) receptor (DP receptor) results in stimulation of adenylyl cyclase, resulting in inhibition of platelet aggregation and smooth muscle relaxation. A second PGD(2) receptor has recently been identified and designated as the DP(2) receptor, or chemoattractant receptor-homologous molecule expressed on Th2 cells. PGD(2) acts through the DP(2) receptor to induce eosinophil chemotaxis, actin polymerization, calcium mobilization, and adhesion molecule expression. The most potent DP(2) receptor agonist yet identified is 15R-methyl-PGD(2), which has the unnatural R configuration at C(15). 15-Deoxy-Delta(12,14)-PGJ(2) is also a potent DP(2) receptor agonist that activates eosinophils at concentrations much lower than those required for its anti-inflammatory effects. Because of its critical location in the lung and its potent effects on eosinophils, PGD(2) may be an important proinflammatory mediator in asthma. PMID- 12895602 TI - The DP receptor, allergic inflammation and asthma. AB - Prostaglandin (PG) D(2) is the major cyclooxygenase metabolite of arachidonic acid produced by mast cells in response to allergen in diseases, such as asthma, atopic dermatitis, allergic rhinitis and allergic conjunctivitis. However, whether PGD(2) regulates allergic process per se, and, if so, whether it facilitates or down-regulates the disease process has remained unknown. PGD(2) exerts its actions by binding to two types of specific cell surface receptor. One is DP (the PGD receptor) and the other is chemoattractant receptor-homologous molecule expressed on Th2. Between the two, the DP receptor has been better characterized since its cDNA cloning in 1994, and novel class of DP antagonists have been and are being developed. Furthermore, mice deficient in DP were generated and have been subjected to several models of allergic diseases to reveal the role of PGD(2) in allergy. In this article, we summarize these findings and provide an overview of the current status of the DP receptor research to discuss the therapeutic potential of modulating the PGD(2)-DP pathway in allergic diseases. PMID- 12895603 TI - The role of mast cells in asthma. AB - While the role of mast cells in allergic reactions is unequivocal, their precise functions in asthma remain controversial. Mast cells uniquely populate all vascularized organs and tissues, including the upper and lower respiratory tree, even in healthy individuals. Histologic evidence suggests that asthma is accompanied by a mast cell hyperplasia in the inflamed mucosal epithelium and the adjacent smooth muscle. The mechanisms responsible for constitutive mast cell development have been partly elucidated. Moreover, both in vitro studies and in vivo disease models indicate that mast cells have a remarkably flexible program of gene expression, and this program can be drastically altered by the T-cell derived Th2 cytokines relevant to asthma. Moreover, the role of mast cells in innate immunity is now firmly established, and the capacity for numerous microbial pathogens to initiate their activation in vitro and in vivo suggest mechanisms by which microbes could initiate disease exacerbations. PMID- 12895604 TI - Effects of preoperative chemoradiotherapy on postsurgical morbidity and mortality in cT3-4 +/- cM1lymph cancer of the oesophagus and gastro-oesophageal junction. AB - OBJECTIVE: Very few studies have examined post-operative morbidity after resection of oesophageal carcinoma, especially in patients treated with induction chemo- and radiotherapy for locally advanced stages. This study assessed the effects of induction chemoradiotherapy on post-operative course after resection of locally advanced oesophageal carcinoma (cT3-4 + cM1lymph). METHODS: Induction therapy consisted of 5-fluorouracil days 1-5 and days 21-25, cisplatin day 1 + day 21 and concomitant radiotherapy 18-20 fractions of 2Gy (total dose 36-40Gy). Induction chemoradiotherapy was completed in 109 patients. Surgery was performed in 90 patients (operability: 90/109 = 83%): 85 patients underwent resection with curative intent (resectability: 85/109 = 78%), bypass operation was performed in five patients. Nineteen patients could not be operated on. Results were compared to a matched group of pT3M1LYM/pT4 patients (n = 86) who underwent primary surgery in the same period. RESULTS: Resection was complete (R0) in 68 patients (68/90 = 76%). Mean duration of surgery was 428 min (range: 240-690). Peroperative complications were haemorrhage in three patients (3/90 = 3.3%), tracheobronchial perforation in three patients (3/90 = 3.3%). Median total hospital stay was 20.5 days (range: 8-355). Mean duration of intubation was 7 days (range: 1-190); 67 patients (67/90 = 74.4%) were intubated for less than 24 h. Non-tumour related hospital mortality after resection was 8.3% (7/84 patients). Mortality after two-field lymphadenectomy was 5.2 versus 11.7% after three-field lymphadenectomy. After primary surgery (n = 86) overall mortality was 2.3% (P = 0.015) and nil after two- and three-field lymphadenectomy (P = 0.011). Medical morbidity consisted of pneumonia in 43 patients (43/90 = 48%), atelectasis in ten patients (10/90 = 11%), dysrhythmia in 21 patients (21/90 = 23%), sepsis in 11 patients (11/90 = 12%) and adult respiratory distress syndrome in ten patients (10/90 = 11%). Surgical morbidity included pleural effusion in 16 patients (16/90 = 18%), tracheal fistula in two patients (2/90 = 2%), chylothorax in two patients (2/90 = 2%) and acute pancreatitis in one patient (1/90 = 1%). Ten patients (10/90 = 11%) had a radiologically confirmed anastomotic leak; however only in four out of them with clinical manifestation; treatment was conservative in all four patients. Major morbidity occurred in 27 patients (27/90 = 30%). Overall rate of morbidity was significantly higher after three-field lymphadenectomy (85%) as compared to two-field lymphadenectomy (68.7%; P = 0.023). CONCLUSIONS: Chemoradiotherapy followed by resection of cT3-4 +/- cM1lymph oesophageal carcinoma is feasible with acceptable mortality. Mortality, however, seems to be significantly higher when compared to a group of pT3M1LYM/pT4 patients who underwent primary surgery (8.3 versus 2.3%; P = 0.015) in the same period in our department. PMID- 12895605 TI - Maximal thymectomy in children with myasthenia gravis. AB - OBJECTIVES: We performed this study to evaluate the benefit of thymectomy in children with myasthenia gravis (MG). METHODS: Over a period of 15 years from 1986 to 2001, we collected data on 30 children with MG and retrospectively reviewed the outcome of maximal thymectomy. RESULTS: There were 23 females and seven males with a mean age of 13.2 years (range 4-16). The mean duration of the disease was 19.3 months (range 2-144). According to Osserman classification, there were 14 children in class II; 12 in class III; and four children in class IV. One child in class IV required postoperative ventilation and one was re explored to drain a pericardial effusion secondary to central line leak. We found ectopic thymic tissue in 10 cases (33.3%). During a mean follow-up period of 53.5 months (range 9-180), complete remission was noted in 13 children (43.4%) and improvement in 14 (46.6%). The remaining three children (10%) did not improve following surgery. Univariate analysis (P < 0.05) showed that ectopic thymic tissue is a significant prognostic factor for outcome. CONCLUSION: Maximal thymectomy appears to provide a high rate of remission and improvement in children with MG. However, the presence of ectopic thymic tissue has poor prognostic value. PMID- 12895606 TI - Video-assisted mediastinoscopic lymphadenectomy (VAMLA)--a method for systematic mediastinal lymphnode dissection. AB - OBJECTIVE: Video-assisted mediastinal lymphadenectomy (VAMLA) increases quality of mediastinal lymph node staging in bronchial carcinoma. The video mediastinoscope allows systematic lymphadenectomy by bimanual preparation. Complete bilateral resection of lymph nodes in stations 1, 2, 3, 4 and 7 (Naruke) can safely be done after visualization of limiting structures (trachea, main bronchi, oesophagus, pericardium, pulmonary artery, aorta, upper vena cava and azygos vein). In this initial study, we compared histopathological findings from VAMLA with final lymph node staging from subsequent thoracotomy. METHODS: Between January 2001 and December 2001, 25 patients were operated by VAMLA (among 162 mediastinoscopies), two patients for diagnostic purposes and 23 for staging of bronchial carcinoma. Eighteen patients underwent subsequent thoracotomy for tumor resection and systematic lymphadenectomy. Pathological findings were reviewed. RESULTS: In VAMLA, lymph node dissection of station 2R, 2L and 4R was achieved in 96, 28 and 92%, respectively, whereas resection of lymph nodes in station 7 and 4L was performed in 100%. Other locations were dissected in 44%. A mean of 8.6 lymph nodes were removed in each patient. No residual lymph node tissue was found in the subcarinal compartment at open surgery. When comparing histopathological staging from VAMLA with final pathology, there were no false negative results. Seventeen patients who had N0 disease at VAMLA proved to be N0 or N1 at thoracotomy, one patient diagnosed as N2 at mediastinoscopy had N2 disease at final pathology. The only complication observed in VAMLA was a blood loss of >100 ml in 12% of patients without need for transfusion or surgical intervention. CONCLUSION: Mediastinal lymph node staging is improved by VAMLA. A systematic lymphadenectomy is performed bimanually through the video mediastinoscope. The number of lymph nodes removed is doubled compared to standard mediastinoscopy. There were no false negative results at final pathology. This new technique presents the basis for video-assisted thoracic surgery (VATS) lobectomy because complete resection of the mediastinal lymph nodes can be achieved by VAMLA. Potential complications of VAMLA such as injury of major mediastinal vessels, airways, pneumothorax or recurrent laryngeal nerve injury indicate the need for a full thoracic surgical infrastructure. PMID- 12895607 TI - Strong additive effect of 1,25-dihydroxycholecalciferol and cyclosporine A but not tacrolimus in rat lung allotransplantation. AB - OBJECTIVES: 1,25-Dihydroxycholecalciferol (calcitriol, vitamin D3) has immunosuppressive properties. This study evaluates the effect of calcitriol in combination with either cyclosporine A or tacrolimus on acute lung allograft rejection in a rat model of unilateral left lung allotransplantation. METHODS: Unilateral left lung transplantation was performed in male rats (Brown-Norway to Fischer F344, 200-250 g body weight). For immunosuppression, the following subtherapeutic doses were used: calcitriol 0.5 microg/kg/day, cyclosporine A 2.5 mg/kg/day i.p., and tacrolimus 40 microg/kg i.m. Five groups (n = 5) were analyzed: cyclosporine A; cyclosporine A and calcitriol; calcitriol; tacrolimus and calcitriol; and tacrolimus. The injections were performed for 5 days starting from the day of transplantation. Recipients were sacrificed on day 5 post transplant. The contralateral right main bronchus and pulmonary artery were occluded for 5 min and blood was drawn for blood gas analysis. The grafts were excised, fixed in formaline and embedded in paraffin. Histological evaluation was done in blinded fashion (ISHLT 1999/rank scale). The mean and standard error of the mean (PaO2) or the median and range (rejection grading) are given. ANOVA followed by planned comparison for the PaO2 and Kruskal-Wallis ANOVA for rejection grading were applied, p < 0.05 considered significant. RESULTS: Arterial PaO2 on day 5 was very low in animals treated with subtherapeutic dosages of either cyclosporine A (48 +/- 10 mmHg), calcitriol (51 +/- 3) or tacrolimus (86 +/- 22). Combined treatment with cyclosporine A and calcitriol revealed a significant improvement (248 +/- 78; p < 0.05 vs. other groups), whereas the combination of tacrolimus with calcitriol did not reveal any benefit (65 +/- 9). Rejection grading with these subtherapeutic doses did not show any significant difference between groups. CONCLUSIONS: Our data indicate that cyclosporine A, but not tacrolimus, has a strong additive effect with calcitriol on acute rat lung allograft rejection. PMID- 12895608 TI - Tissue engineered cartilage generated from human trachea using DegraPol scaffold. AB - OBJECTIVE: To date numerous attempts have been undertaken to conquer the challenging problem of reconstructing long segmental tracheal defects, as yet without lasting success. Recently, employing concepts of tissue engineering in animals, cartilage-like constructs were transplanted in vivo. However, both the feasibility of fabricating tracheal replacements and the use of human tracheal chondrocytes (HTC) for tissue engineering are still under investigation. In this study, we optimized isolation and cultivation techniques for human tracheal cartilage, assessing the feasibility of seeding these cells onto a novel, three dimensional (3-D) polyester-urethane polymer (DegraPol). METHODS: Human tracheal cartilage was harvested from the trachea of lung donors, digested in 0.3% collagenase II, and the condrocytes serially passaged every 7-9 days. Cells were also cultivated over agar plate during the total 6-8 weeks expansion phase. Thereafter, chondrocytes were seeded onto DegraPol (pore sizes 150-200 microm) with a seeding density of 2.4 x 10(7)/ml, and chondrocyte-polymer constructs maintained during in vitro static culture. RESULTS: HTC displayed stable proliferation kinetics in monolayer culture with positive expression of collagen type II. Following polymer seeding, both cellular proliferation and extracellular matrix (ECM) production, as measured by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide) and glycosaminoglycan assays, continued over extended culture. Active growth of HTC on DegraPol was further demonstrated by Alcian blue staining, with the histomorphological appearance of the construct resembling that of native cartilage. Scanning electron microscopy showed chondrocyte growth and ECM synthesis both on the surface and inside the porous scaffold, with a dense cell layer on the surface of the scaffold and a lower cell distribution in the scaffold's interior. CONCLUSIONS: The harvested chondrocytes from human trachea cartilage expand well in vitro and possess the ability to form new cartilage-like tissue when seeded onto DegraPol matrix. However, improved culture conditions are needed to permit cellular growth throughout cell-polymer constructs. PMID- 12895609 TI - Use of self-expanding vascular endoprostheses in superior vena cava syndrome. AB - INTRODUCTION: Superior vena cava syndrome (SVCS) is associated to a malignant tumor in more than 90% of cases; being the lung cancer the most frequent (80%). SVCS has a benign cause in less than 5% of cases. Endovascular stenting has been proposed as the primary treatment of choice. We report our experience in SVC recanalization through the use of self-expanding vascular stents as treatment of life-threatening SVCS of benign and malignant etiology. MATERIALS AND METHODS: Between January 1994 and April 2002 44 patients with critical SVCS, were treated at the Hospital Italiano de Buenos Aires. Forty nine self-expanding endovascular metallic stents were percutaneously placed in the SVC. Thirty-one (70%) patients were male and 13 (30%) were female. The mean age was 55.6 years (range: 21-77). The etiology of SVCS was malignant in 40 cases and benign in 4. The malignant causes included lung cancer: 37 (37/44 - 92.5%), lymphoma: 1 (2.5%), chondrosarcoma 1 (2.5%), melanoma 1 (2.5%). The benign etiology corresponded to central catheters (N: 2) and post-radiation fibrosis (N: 2). Cavography showed complete occlusion of SVC in 12 cases (27%) and significant partial stenosis in 32 cases (73%). Thrombi associated with tumor stenosis were present in 25 (57%) patients. RESULTS: All procedures were technically successful. No stent migration was observed. Thirty-two patients with malignant tumor ultimately died due to the progression of the disease. Mean survival time was 193 days (range: 25-578). SVCS recurrence was observed on six occasions. In four patients a new stent was placed. Symptomatic improvement was dramatically seen within 24-48 h after stent placement in 40 patients (90.9%) and 83.3% out of the cases (38/44) were symptoms free during the rest of the disease. Three patients died in the 7 following days. CONCLUSION: The use of self-expanding vascular endoprostheses in the recanalization treatment of SVC in SVCS due to a malignant or benign etiology offers excellent results with rapid and prolonged remission of symptoms. PMID- 12895610 TI - Self-expandable valved stent of large size: off-bypass implantation in pulmonary position. AB - OBJECTIVE: To evaluate the feasibility of the off-bypass implantation of a self expandable valved stent of large size in pulmonary position. MATERIALS AND METHODS: A glutaraldehyde preserved valved bovine jugular xenograft with internal diameter = 22 mm, mounted in two rings of nitinol 'Z' stent, expandable from 7 to 24 mm of internal diameter, was acutely evaluated in 6 adult pigs, mean body weight 55.6 kg (range 47-67 kg). Through a stent-graft delivery system (24 French) the self expandable valved stent was implanted off-bypass in pulmonary valve position by trans-ventricular approach through median sternotomy. RESULTS: The mean diameter of the main pulmonary artery measured was 21.7 +/- 1.6 mm. The mean length of the self expandable valved stent was 23.1 +/- 0.7 mm, the mean internal diameter 21.6 +/- 0.7 mm and the mean external diameter 26.3 +/- 0.7 mm. The mean peak pressure gradient recorded across the valve was 6.33 +/- 2.8 mmHg (range 4.5-9.6 mmHg) at Doppler echocardiography, and 4.5 +/- 3.1 mmHg (range 0-7 mmHg) at invasive measurement, with a pulmonary blood flow of 3.03 +/- 0.05 l/min. Intra-vascular ultrasound showed complete opening and closure of the valve (mean area reduction from 315.08 +/- 54.13 to 0 mm2). CONCLUSIONS: (a) Off-bypass implantation of self-expandable valved stent is feasible in pulmonary position; (b) off-bypass surgical approach allows for valved stent implantation of adult size with adequate hemodynamic functioning; and (c) intra-vascular ultrasound makes implantation and evaluation easy and reproducible. PMID- 12895611 TI - Does moderate mitral regurgitation impact early or mid-term clinical outcome in patients undergoing isolated aortic valve replacement for aortic stenosis? AB - OBJECTIVE: The early and mid-term impact of functional mitral regurgitation (MR) in patients undergoing isolated aortic valve replacement (AVR) for aortic stenosis remains unresolved. METHOD: Through our institutional databank, using a case-match study, we identified 58 patients with MR grades 0-1 and 58 patients with MR grades 2-3 (patients matched for sex, age, ejection fraction (EF), NYHA, diabetes, and CVA). Data were collected prospectively (mean duration of follow up: 3.2 +/- 2.4 years). RESULTS: Perioperative morbidity (re-operation for bleeding, low cardiac output, CVA, renal failure) was comparable among groups. Difference in mortality between the two groups was non-significant (7.0 vs. 3.5%, P = 0.67 in groups MR 2-3 vs. 0-1, respectively). At early echocardiographic follow-up, 7/58 patients (12.1%) within group MR grades 0-1 increased their MR to grades 2-3; among which only two remained with MR grades 2-3 at mid-term follow up. Within MR group 2-3, 18/58 (31.0%) remained with MR grades 2-3 among which 7/18 (38.9%) decreased of at least one grade at follow-up. Eight year actuarial survival was comparable in both groups: MR grades 0-1 = 60.9% vs. MR grades 2-3 = 55.0%; P = 0.1. Actuarial survival of patients with MR grades 2-3 postoperatively was similar to patients with MR grades 0-1 (MR grades 0-1 = 59.0%, MR grades 2-3 = 58.9%, P = NS). CONCLUSIONS: Presence of preoperative moderate functional MR (grades 2-3) in patients undergoing isolated AVR for aortic stenosis regresses in the majority of patients postoperatively and has no significant impact on perioperative morbidity or mortality, nor mid-term survival. Thus, moderate functional MR should be treated conservatively in the majority of patients especially in the elderly subjected to isolated AVR for aortic stenosis. PMID- 12895612 TI - Combined radiofrequency modified maze and mitral valve procedure through a port access approach: early and mid-term results. AB - OBJECTIVE: The aim of this study was to assess the feasibility and effectiveness of irrigated radiofrequency (RF) modified Maze procedure through a port access approach during mitral valve surgery and evaluate early and mid-term results. MATERIAL AND METHOD: During a 16 months time period, 67 patients with chronic atrial fibrillation (AF) eligible for port access mitral valve surgery were randomly assigned to either Group A, in which they underwent a combined procedure (N = 33) or Group B, in which a valve procedure alone was performed (N = 34). Both groups were similar in terms of age, sex, valve pathology, duration of AF left atrial diameter and left ventricle function (P > 0.05). Four had undergone previous operations. RESULTS: Median follow-up was 10 months for both groups, 95% CI (9.18-10.8). One patient in each group died early postoperatively (3 and 2.9%). Two patients required reoperation for bleeding, one in each group (3 and 2.9%). There were two conversions to right thoracotomy. In Group A, freedom from AF was 100% at the end of the operation (76% sinus, 24% pacemaker) Six and twelve months freedom from AF was 87.2 and 93.6%, respectively. In Group B, freedom from AF at the end of operation was 41%. At the end of 6 and 12 months, freedom from AF was 9.4% (P = 0.0001). One patient in Group A required a permanent pacemaker (3%). During follow-up, one patient in Group A died of non-cardiac causes (3%). In Group B, there were two late deaths: one cardiac (2.9%) and one neurologic (2.9%). There were no thromboembolic events detected in Group A during follow-up, whereas two patients in Group B suffered this complication (6%, P = 0.081). At 12 months, functional capacity had improved for patients in both groups (P < 0.0001). CONCLUSION: The combination of mitral valve surgery and irrigated RF Maze procedure was safe and efficient through a port access approach. There were no procedure related complications like esophageal or coronary artery injury. Early and mid-term results were favourable with 93.6% of patients free of AF at 1 year in comparison to the 9.4% of the control group. The data is not sufficient to reach any conclusions in terms of thromboembolic rates, despite favourable results for the RF Maze group. Nevertheless, in terms of feasibilty, sinus rhythm restoration and overall outcome, early results are encouraging and we advocate the use of the combined procedure through a port access approach. PMID- 12895613 TI - Cannulation of the right axillary artery for surgery of acute type A aortic dissection. AB - OBJECTIVE: The optimal choice of the arterial inflow site during operations for type A aortic dissection is not clearly defined. The aim of the prospective study was to identify whether cannulation of the right axillary artery instead of the femoral artery may improve the results of surgery for acute type A aortic dissection. METHODS: Seventy consecutive patients were operated on because of acute type A aortic dissection from January 2000 to February 2002. The only difference in surgical strategy was the site of arterial cannulation: the right axillary artery was used in 20 patients [axillary group] and the left femoral artery in 50 patients [femoral group]. All patients had aortic surgery with open distal anastomosis during deep hypothermic arrest and retrograde cerebral perfusion. The mean age was 58.7 +/- 12 years with a range from 28 to 88 years (axillary group, 56.6 +/- 13 years; femoral group, 59.4 +/- 12 years; P = 0.435). Preoperatively evident organ malperfusion was identified in five (25%) patients of the axillary group and in seven (14%) of the femoral group. RESULTS: There was no perioperative death. The hospital mortality rate was 5.0% for the axillary group and 22% for the femoral group (all patients, 17%). Major neurological complications occurred postoperatively in 5% of patients from the axillary group (one out of 20 patients) and in 8% of patients from the femoral group (four out of 50 patients) (all patients, 7%). CONCLUSION: Cannulation of the right axillary artery improved the outcome of surgery for acute type A aortic dissection. However, postoperative complications occurred after both axillary and femoral artery cannulation. PMID- 12895614 TI - Intramural hematoma and dissection involving ascending aorta: the clinical features and prognosis. AB - OBJECTIVE: The clinical features and remedies of acute aortic intramural hemorrhage (IMH) are well discussed. This study prospectively analyzes the features compared with those of Type A aortic dissection, and evaluate the treatment modalities and the prognosis with Type A IMH managed by our original program, Eighty-six consecutive patients consisted of acute type A IMH (n = 36) and dissection (n = 50) were diagnosed between January 1994 and March 2002. Patients with IMH were older (mean 67 and 60, P = 0.0017), more hypertensive (P = 0.0015), not hyperlipidemic (P = 0.0042) than those with dissection. The incidences of preoperative pericardial effusion and aortic regurgitation were significantly lower in patients with intramural hematoma than with dissection, respectively (8:28 versus 22:28, P = 0.0366, 4:32 versus 22:28, P = 0.0011). METHODS: Urgent operation was performed for the patients of IMH with cardiac tamponade or rupture and all dissections. Uncomplicated patients of the patients with IMH were treated medically. Late surgical conversion was applied for the medical treated case on any condition with persistent pain, progression to type A dissection, ruptured aneurysm, or aneurysmal enlargement (>60 mm). Operative mortality, late cardiovascular event, and long-term survival were evaluated statistically. RESULTS: Ten urgent surgical repairs were performed with type A IMH patients and one patient died postoperatively. The rest 26 patients were treated medically. The mean follow up period was 39 +/- 28 months. Among the 26 patients, seven were converted surgical intervention. Cardiovascular event free curve on the 26 patients (Kaplan-Meier, CI: 95%) was 65.6% (45.9-85.3), 59.1% (37.5-80.6) at 2, 4 years. There were six dissection and six IMH patients death during follow up. Two of IMH patients died from cardiovascular event. The actuarial survival rate (Kaplan-Meier, CI: 95%) was 87.5% (76.0-99.1):87.9% (66.2 97.1), 81.7% (66.2-97.1):87.9% (78.8-97.0) at 2, 4 years (P = 0.8393). CONCLUSIONS: Type A IMH tends to occur in older, more hypertensive and not hyperlipidemic patients, showed lower incidences of preoperative aortic valve regurgitation and pericardial effusion than dissection. Medical treatment alone was not enough to manage all type A IMH patients, and 47.2% (17/36) of the patients needed surgical intervention. Urgent surgical repair was not necessary for all type A IMH patients to achieve favorable surgical outcome with careful follow-up using imaging modality. PMID- 12895615 TI - Modified ultrafiltration may not improve neurologic outcome following deep hypothermic circulatory arrest. AB - OBJECTIVE: Modified ultrafiltration (MUF) improves systolic blood pressure and left ventricular performance, as well as lowering transfusion requirements, after cardiopulmonary bypass (CPB). MUF has also been shown to enhance acute cerebral metabolic recovery after deep hypothermic circulatory arrest (DHCA), but whether this improves neurologic outcome is unknown. METHODS: Sixteen neonatal piglets underwent CPB and 90 min of DHCA. The hematocrit was maintained between 25 and 30%. Alpha-stat blood gas management was used. After separation from CPB, animals were randomized to 15 min of MUF (n = 8) or no intervention (n = 8). Neurologic injury was assessed with behavior scores and histologic examination. Standardized behavior scores were obtained on post-operative days 1, 3, and 6 (0 = no deficit to 95 = brain death). The percentage of injured neurons by hematoxylin and eosin staining and the degree of reactive astrocytosis by glial filbrillary acidic protein (GFAP) immunohistochemistry were assessed to determine histologic scores in the neocortex and hippocampus (0 = no injury to 4 = diffuse injury). RESULTS: There were no statistically significant differences between groups during CPB. After MUF, the hematocrit was significantly higher (40% +/- 5.7 vs. 28% +/- 3.9, P < 0.001). There were no significant differences in behavior scores between groups (p > 0.1). There was resolution of deficits by day 6 in all animals. Neuronal injury was present in 81% (13/16) of the animals with no statistically significant differences between groups in incidence or severity. CONCLUSIONS: Use of MUF after DHCA does not prevent neuronal injury or improve neurologic outcome in this neonatal swine model. PMID- 12895616 TI - Surgery of the dilated aortic root and ascending aorta in pediatric patients: techniques and results. AB - OBJECTIVE: Dilatation of the aortic root is a well-known cardiovascular manifestation in children and adult patients with connective tissue disease (e.g. Marfan syndrome). Dilatation of the ascending aorta is extremely rare and may be associated with bicuspid aortic valve. This report evaluates the incidence of dilatative aortic root and ascending aortic pathology in patients younger than 18 years and analyzes the results obtained after repair and replacement strategies. METHODS: Between 1/1995 and 12/2002, a total of 752 operations on the thoracic aorta were performed in adult and pediatric patients. We present our experience with a group of 26 patients <18 years of age, who required isolated surgery of the aortic root and/or ascending aorta because of a dilatative lesion. Fifteen patients had isolated aortic root dilatation (13 of them suffered from Marfan syndrome), eight patients presented with an idiopathic dilatation of the ascending aorta and three patients had dilatation in association with a bicuspid aortic valve. Mean age was 10 +/- 4.8 years (4-18 years). Repair of the aortic root with preservation of the aortic valve (Yacoub, David or selective sinus repair) was performed in nine patients, replacement using a homograft was performed in five patients, composite graft with mechanical prosthesis in two patients, with biological prosthesis in one patient and Ross operation was performed in one case. Isolated supracoronary graft replacement was performed in eight patients. RESULTS: Two patients died during hospitalization: a 10-year old girl developed respiratory failure on the 2nd postoperative day and autopsy revealed Ehlers-Danlos syndrome with a massive intrapulmonary emphysema. A 14 year-old Marfan patient with severely depressed preoperative LV function died from low cardiac output following composite-graft, mitral and tricuspid valve repair. One patient required aortic valve replacement 7 days after an aortic valve sparing root repair. There was no additional perioperative morbidity. In the long-term, two patients died from rupture of the thoracic aorta, both following minor non-cardiovascular surgical procedures. Both had normal sized descending and abdominal aorta. CONCLUSION: Repair of the aortic root and/or ascending aorta in children and adolescent patients can be performed with acceptable early and late results. While the presence of severe comorbidity may adversely affect early outcome, long-term survival was mainly determined by rupture of the descending aorta. PMID- 12895617 TI - Surgical reinterventions following the Fontan procedure. AB - OBJECTIVE: The Fontan procedure is utilized as a final reconstructive procedure for patients with functional single ventricle. Short- and long-term outcomes have improved significantly, however, some patients require additional cardiac procedures following the Fontan operation. The outcomes for these reinterventions are not known. METHODS: Cardiac Surgery and Cardiac Intensive Care Unit databases at The Children's Hospital of Philadelphia were reviewed to identify all patients who underwent cardiac surgery after a previous Fontan operation between January 1, 1995 and December 31, 2001. RESULTS: During the study period, 123 procedures were performed in 71 patients. The median time from Fontan to reoperation was 3.6 years (range 0.1-20 years). Indications for reintervention included arrhythmia, cyanosis, 'failing' Fontan circulation or exercise intolerance, protein losing enteropathy, atrioventricular valve (AVV) regurgitation, and other indications. Procedures included pacemaker insertion or revision (n = 59, 48%), reinclusion of previously excluded hepatic veins (n = 16, 13%), revision to either a lateral tunnel or extra-cardiac conduit Fontan (n = 13, 11%), cardiac transplantation (n = 9, 7%), enlargement or creation of a baffle fenestration (n = 6, 5%), isolated AVV repair or replacement (n = 2, 2%), and other procedures (n = 18, 14%). There were five early and five late deaths. Hospital mortality was greatest for patients undergoing cardiac transplantation (4/9, 44%), accounting for 80% of the early deaths. CONCLUSIONS: Surgical reinterventions following the Fontan procedure may be necessary for multiple indications which result in impairment of the Fontan circulation. Most reinterventions can be performed with minimal morbidity and mortality. Survival for patients requiring cardiac transplantation following the Fontan procedure remains poor. PMID- 12895618 TI - Biological effects of off-pump vs. on-pump coronary artery surgery: focus on inflammation, hemostasis and oxidative stress. AB - Cardiopulmonary bypass (CPB) has been recognized as a cause of complex systemic inflammatory response, which significantly contributes to several adverse postoperative complications. In the last few years, off-pump coronary artery bypass grafting has gained widespread diffusion as an alternative technique to conventional on-pump coronary artery bypass grafting. Surgeons supporting off pump surgery state that the avoidance of the CPB and myocardial ischemia reperfusion significantly reduces the postoperative systemic inflammatory response and other biological derangements and, possibly, may improve the clinical outcomes. We review, here, the available evidence concerning possible differences between off-pump and on-pump procedures in terms of inflammation, hemostasis and oxidative stress. Consistent differences in the involvement of these systems are observed, but they are limited to the final steps of the surgical procedures and the early hours after. These findings suggest that the global surgical trauma may be as important, or even more, as the CPB in terms of systemic inflammatory and coagulation-fibrinolytic pathway activation. Further studies are needed in order to confirm this hypothesis. PMID- 12895619 TI - A validated rule for predicting patients who require prolonged ventilation post cardiac surgery. AB - OBJECTIVE: Prolonged ventilation post surgery causes logistic problems on cardiac surgical intensive care units (CSU). We thus sought to derive and validate a clinical decision rule to predict patients at high risk of prolonged ventilation, so that the timing of operations on high risk patients can be optimised in the context of the workload of the CSU. METHODS: The North Staffordshire Royal Infirmary (NSRI) Open Heart Registry was analysed from April 1998 to May 2002. Prolonged ventilation was defined as that which was longer than 24 h. The Parsonnet score was assessed for its ability to predict these patients. Univariate analysis was first performed to identify predictive variables. Recursive partitioning and logistic regression was then performed to identify the optimal decision rule. This rule was then validated on the Blackpool Victoria Hospital (BVH) Open Heart Registry. RESULTS: A total of 3,070 patients were analysed of whom 201 were ventilated for more than 24 h. A Parsonnet score of 10 predicted 49% of high risk patients but 618 low risk patients are misclassified. Our rule that uses Parsonnet score over 7, ejection fraction, operation status, PA pressure and age, to identify high risk patients identifies 50% of those needing prolonged ventilation and only incorrectly identifies 282 of the 2869 patients with normal ventilation times giving a specificity of over 90%. Validation in the BVH database demonstrated similar findings. CONCLUSION: Our rule identifies 14% of all our patients as high risk and 50% of these required prolonged ventilation. Such a rule allows more efficient use of scarce CSU resources by appropriate surgical scheduling. PMID- 12895620 TI - Intra-aortic balloon pumping: effects on left ventricular diastolic function. AB - OBJECTIVE: The intra-aortic balloon pump is the most widely used form of temporary cardiac assist and often utilised in patients before and after cardiac surgery. Several effects of balloon counter-pulsation have been reported previously, but its effect on left ventricular diastolic function has not been thoroughly investigated. The aim of this study is to examine the effect of the intra-aortic balloon pump on left ventricular wall motion and transmitral flow. METHODS: We studied 20 patients in the intensive care unit, less than 36 h following cardiac surgery. We recorded left anterior descending coronary artery and transmitral E-wave flow velocities using transesophageal echocardiography pulsed Doppler. We also recorded left ventricular long axis free-wall movement using M-mode. The intra-aortic balloon pump was set to full augmentation and recordings were made at pumping cycles 1:1, 1:2, 1:3, and when the pump was on stand-by, leaving a minimum of 5 min between the pumping modes to allow the return to control conditions. In order to eliminate time effects, the sequence of recording was varied between patients using a 4 by 4 Latin-square. RESULTS: The peak diastolic left anterior descending coronary artery and transmitral E-wave flow velocities, and left ventricular free-wall early diastolic lengthening velocity increased significantly with intra-aortic balloon pumping cycles 1:1, 1:2 and 1:3 compared to their value with the pump on stand-by, all P < 0.001. The increase in peak transmitral E-wave flow velocity correlated with the increase in peak left anterior descending coronary artery diastolic flow velocity (r = 0.74, P = 0.02), and with the increase in left ventricular free-wall early diastolic lengthening velocity (r = 0.80, P < 0.001). CONCLUSION: Using the intra-aortic balloon pump post-cardiac surgery significantly increases peak diastolic left anterior descending coronary artery flow velocities and left ventricular free wall early diastolic lengthening velocity, whose increase explains the increase in peak transmitral E-wave velocity. Although coronary flow is epicardial and mitral flow is intracardial, their close relationship suggests an improvement in left ventricular diastolic function with intra-aortic balloon pump. PMID- 12895621 TI - Long-term survival (>10 years) of patients >60 years with induction therapy after cardiac transplantation. AB - OBJECTIVE: Cardiac transplantation has become an established method for end-stage heart disease. Short- and mid-term outcome has been known to be similar between younger and older (>60 years) recipients. So far, nothing is known about long term outcome of old patients and the potential long-term effects of antibody induction therapy in these patients. The purpose of this study was to analyse long-term outcome of old cardiac transplant recipients who underwent antibody induction therapy. METHODS: Since 1989, 203 patients (total n = 882) above 60 years have been transplanted at our center. On these patients n = 66 were above 65 years. Survival, incidences of rejection, infection, cancer, graft arteriosclerosis and the amount of renal insufficiency were compared with patients <60 years (n = 679), transplanted during the same period of time. Freedom from specific event was computed by Kaplan-Meier analysis and compared by log-rank test. RESULTS: Ten year survival was similar in all groups (<60 years: 53.7%; 60-64 years: 53.1% and >65 years: 60.2%; P = NS). Causes of death were similar in all patient groups. There were significant fewer rejection episodes in the older patient group (freedom from rejection: 74.9 vs. 83.5 vs. 90.6; P = 0.03). Yet significantly more number of patients >65 years were without steroid maintenance therapy (43.1%) compared to other patient groups (8.2 vs. 9.3%; P < 0.05). There was no difference in overall freedom from severe infection (74.1 vs. 67.7 vs. 85.3%; P = NS), whereas there was a trend towards more CMV disease in the oldest patient group (82.7 vs. 88.6 vs. 70.8%; P=0.06). The incidence of cancer was similar in all groups (freedom from cancer: 82.2 vs. 84.7 vs. 79.1%; P = NS), as well as there was no difference in severe graftsclerosis between all patients (79.2 vs. 93.7 vs. 93.3%; P = NS). There was no difference in development of chronic renal dysfunction (creatinine > 2.0 mg/dl) between the three groups (10 vs. 14 vs. 16%; P = NS). CONCLUSIONS: Old recipients of cardiac transplants have a similar long-term outcome than younger recipients. They were less prone to rejections, had a similar incidence of severe infections and showed a trend towards more CMV disease. All patients had a very low rate of graft arteriosclerosis that was similar amongst the groups. Age-related decline of the immune system further enhanced by immunomodulation of antibody induction therapy might be accounted for the results as well as steroid-free immunosuppression. PMID- 12895622 TI - Preservation of myocyte structure and mitochondrial integrity in subzero cryopreservation of mammalian hearts for transplantation using antifreeze proteins--an electron microscopy study. AB - OBJECTIVE: Freeze tolerant fish and insects in nature are able to survive subzero temperatures by noncolligatively lowering the freezing temperature of their body fluids using a family of thermal hysteresis proteins (antifreeze proteins, AFPs) specific for each species. Past efforts to cryopreserve mammalian hearts using these proteins were unsuccessful. We report the first successful subzero cryopreservation of rat hearts using fish derived antifreeze proteins with preservation of myocyte structure. METHODS: Heterotopic heart transplantations were performed in isoimmunic Sprague Dawley rats. Donors' hearts were arrested using University of Wisconsin (UW) solution and preserved in UW solution containing AFP I (six experiments) or AFP III (six experiments) at concentrations of 15-20 mg/cc for 2-6 h at subzero temperatures ranging from -1.1 to -1.3 degrees C. Four control experiments were performed by preserving harvested hearts in UW solution alone at -1.3 degrees C for 6 h. In all experiments ice was added in the solution for crystallization. Heterotopic transplantations were performed in the abdomen of the recipient rats. Viability was visually assessed and graded on a scale of 1 (poor contraction) to 6 (excellent contraction). The hearts were then fixed in vivo and processed for electron microscopy study. RESULTS: All hearts preserved at subzero temperatures using AFP I or AFP III survived displaying viability scores of 4-6 1 h after transplantation. Three of the four control hearts that were preserved at -1.3 degrees C without the protective effect of AFP froze and died upon reperfusion. Electron microscopy study of hearts preserved with AFP demonstrated preservation of myocyte structure and mitochondrial integrity. CONCLUSION: Subzero cryopreservation of mammalian hearts for transplantation using AFP I or AFP III is feasible with preservation of myocyte structure and mitochondrial integrity. PMID- 12895623 TI - Echocardiographic diagnosis of incomplete St. Jude's bileaflet valvular closure after mitral valve replacement with subvalvular preservation. PMID- 12895624 TI - Ruptured ascending aorta aneurysm presenting as pulmonary embolism. PMID- 12895625 TI - Intrathoracic gossypiboma: a movable body within a pseudocystic mass. PMID- 12895626 TI - Foreign body aspiration: where can it be? PMID- 12895627 TI - Dyspnea due to giant goiter. PMID- 12895628 TI - Metachronous triple cancer: esophageal carcinoma 4 years later the synchronous bilateral bronchogenic carcinoma. PMID- 12895629 TI - Asymptomatic post-pneumonectomy over-expansion of the residual lung. PMID- 12895630 TI - Ossification of an intercostal flap after carinal resection: long-term concerns of a 'protection maneuver'. PMID- 12895631 TI - New experience with the paracardial right ventricular axial flow micropump impella elect 600. AB - Beating heart surgery is still challenging and could be made safer using assisted circulation. We evaluated clinical outcome following beating heart myocardial revascularisation using the impella elect system with intracardial left ventricular (LV) and paracardial right ventricular (RV) microaxial pumps in eight patients. Clinical course was uneventful. There were no device-related complications. The paracardial design of the RV-pump is a great advantage compared to the former intracardial design because it eliminates arrhythmia and LV hypovolemia. Support by microaxial pumps can make beating heart surgery safer and applicable for more complex cases. PMID- 12895632 TI - Resection of the ascending aorta and aortic valve patch closure for type A aortic dissection after Novacor LVAD insertion. AB - A 60-year-old patient developed an acute type A aortic dissection in the postoperative course of a Novacor left ventricular assist device. We performed a resection of the ascending aorta with an aortic valve patch closure, end-to-end anastomosis of the outflow graft to the distal ascending aorta and two venous grafts to the coronary arteries, in order to avoid residual aortic insufficiency and bleeding related to exposure of the fragilized tissues to high pressures. PMID- 12895633 TI - A rare case of aortocoronary dissection following percutaneous transluminal coronary angioplasty: successful treatment using off-pump coronary artery bypass grafting. AB - Retrograde dissection of the aorta is a rare but potentially life threatening complication of percutaneous coronary intervention. We describe a case of retrograde aortic dissection, which occurred during attempted percutaneous transluminal coroanary angioplasty (PTCA) of chronically occluded left anterior descending artery (LAD). Emergency coronary artery bypass grafting (CABG) was performed to revascularise LAD distal to the site of entry tear with pedicled left internal mammary artery (LIMA). This was achieved off-pump. Ascending aorta was not replaced as the dissection was localised. PMID- 12895634 TI - Acute aortic dissection after off-pump coronary artery surgery. AB - Iatrogenic aortic dissection has been described, albeit infrequently, after coronary artery surgery performed under cardiopulmonary bypass. Since the advent of beating heart coronary surgery, several authors have described an apparent increase of this complication related to the application of a lateral clamp on the ascending aorta to perform the proximal anastomosis. We describe the case of a 70 years old patient who presented aortic dissection, with immediate paraplegia, 7 days after off-pump coronary surgery. PMID- 12895635 TI - Ruptured sinus of Valsalva aneurysm associated with aortic regurgitation caused by hemodynamic effect solely. AB - We describe a case of ruptured sinus of Valsalva aneurysm (RSVA) with moderate aortic regurgitation (AR), which developed on the second day after admission. The AR was caused by a hemodynamic effect solely, in which the shunt blood flow through ruptured site pulled the right aortic cusp away from closure. The pathological mechanism of the AR was clearly visualized by intraoperative transesophageal echocardiography (TEE) and the AR was successfully resolved after simple closure of the RSVA without any additional procedure to the aortic valve. PMID- 12895636 TI - Successful treatment of Salmonella mycotic aneurysm of the descending thoracic aorta. AB - Salmonella mycotic aneurysms of the descending thoracic aorta are exceedingly rare. There are few case reports and even fewer reports of long term survival. The case of a 68-year-old female presenting with a mycotic aneurysm of the descending thoracic aorta caused by Salmonella species is described, which involved successful surgical intervention. PMID- 12895637 TI - Right ventricular inflow obstruction from massive fungal vegetation presenting as neonatal circulatory collapse. AB - Occurrence of neonatal circulatory collapse imposes effective differential diagnosis and expeditious therapeutic intervention. We report a case of neonatal cardiogenic shock, caused by a massive intra-cardiac fungal vegetation. PMID- 12895638 TI - Dramatic post-cardiotomy outcome, due to severe anaphylactic reaction to protamine. AB - Immunologic reactions to protamine sulfate during cardiac surgery are very rare. The frequency and outcome of such adverse reactions is unclear. We report a case of lethal anaphylactic reaction to protamine that occurred in a non-diabetic patient following the uneventful replacement of the ascending aorta. We also briefly review the mechanisms of this adverse reaction and emit some considerations on the management of this situation. PMID- 12895639 TI - Pulmonary arteriovenous malformation treated by lobectomy. AB - Pulmonary arteriovenous malformation (PAVM) may occur primarily or in association with hereditary hemorrhagic telangiectasia. We present a case of PAVM in the central lower lobe of the left lung of a 75-year-old woman, which was successfully treated by lobectomy. Contrast echocardiography is an excellent tool for evaluation of this uncommon lesion. Advances in interventional radiology have led to the introduction of obliterative techniques for the treatment of PAVM. However, in the presence of a large solitary malformation centrally located, as in our case, and in high-risk patients, surgery is still a safe and effective first option. PMID- 12895640 TI - Extended operation for invasive thymoma with intracaval and intracardiac extension. AB - Two cases of invasive thymoma with intracaval and intracardiac extension into the right atrium are reported. Radical excisions and reconstructions of the superior vena cava (SVC) requiring extracorporeal circulation were performed. Invasive thymoma with this growth pattern is extremely rare, and patients with SVC obstruction should be evaluated for the resection of tumors and reconstruction of the SVC. PMID- 12895641 TI - Synchronous bilateral lung carcinoid tumors: a rare entity? PMID- 12895643 TI - Prospectively randomized evaluation of stented versus stentless aortic valve bioprostheses. PMID- 12895645 TI - Does the EuroSCORE perform well on the STS population? PMID- 12895647 TI - A retroviral-derived peptide phosphorylates protein kinase D/protein kinase Cmu involving phospholipase C and protein kinase C. AB - CKS-17, a synthetic peptide representing a unique amino acid motif which is highly conserved in retroviral transmembrane proteins and other immunoregulatory proteins, induces selective immunomodulatory functions, both in vitro and in vivo, and activates intracellular signaling molecules such as cAMP and extracellular signal-regulated kinases. In the present study, using Jurkat T cells, we report that CKS-17 phosphorylates protein kinase D (PKD)/protein kinase C (PKC) mu. Total cell extracts from CKS-17-stimulated Jurkat cells were immunoblotted with an anti-phospho-PKCmu antibody. The results show that CKS-17 significantly phosphorylates PKD/PKCmu in a dose- and time-dependent manner. Treatment of cells with the PKC inhibitors GF 109203X and Ro 31-8220, which do not act directly on PKD/PKCmu, attenuates CKS-17-induced phosphorylation of PKD/PKCmu. In contrast, the selective protein kinase A inhibitor H-89 does not reverse the action of CKS-17. Furthermore, a phospholipase C (PLC) selective inhibitor, U-73122, completely blocks the phosphorylation of PKD/PKCmu by CKS-17 while a negative control U-73343 does not. In addition, substitution of lysine for arginine residues in the CKS-17 sequence completely abrogates the ability of CKS-17 to phosphorylate PKD/PKCmu. These results clearly indicate that CKS-17 phosphorylates PKD/PKCmu through a PLC- and PKC-dependent mechanism and that arginine residues play an essential role in this activity of CKS-17, presenting a novel modality of the retroviral peptide CKS-17 and molecular interaction of this compound with target cells. PMID- 12895648 TI - Purification and characterization of a ubiquitin-like peptide with macrophage stimulating, antiproliferative and ribonuclease activities from the mushroom Agrocybe cylindracea. AB - A peptide, with a molecular mass of 9.5kDa and demonstrating an N-terminal sequence similar to ubiquitin, was isolated from fruiting bodies of the mushroom Agrocybe cylindracea. The peptide was isolated with a purification protocol involving ion exchange chromatography on DEAE-cellulose, affinity chromatography on Affi-gel blue gel, FPLC-ion exchange chromatography on Mono S and FPLC-gel filtration on Superdex 75. The peptide was unadsorbed on DEAE-cellulose and adsorbed on Affi-gel blue gel and Mono S. It showed antiproliferative activity on leukemia cell line (M1) and hepatoma cell line (HepG2), and enhanced nitric oxide production in murine peritoneal macrophages with a potency comparable to that of lipopolysaccharide. A pH of 6.0 was required for optimal RNase activity. Its RNase activity was stable over the temperature range of 0-60 degrees C. It exerted ribonucleolytic activity preferentially on polyC, much lower activity on polyU, and negligible activity on polyA and polyG. PMID- 12895649 TI - Peptides of the liver stage antigen-1 (LSA-1) of Plasmodium falciparum bind to human hepatocytes. AB - Synthetic peptides from the liver stage antigen-1 (LSA-1) antigen sequence were used in HepG2 cell and erythrocyte binding assays to identify regions that could be involved in parasite invasion. LSA-1 protein peptides 20630 ((21)INGKIIKNSEKDEIIKSNLRY(40)), 20637 ((157)KEKLQGQQSDSEQERRAY(173)), 20638 ((174)KEKLQEQQSDLEQERLAY(190)) and 20639 (191KEKLQEQQSDLEQERRAY(207)) had high binding activity in HepG2 assays. Were located in immunogenic regions; peptide cell binding was saturable. Peptide 20630 bound specifically to 48kDa HepG2 membrane surface protein. LSA-1 peptides 20630 ((21)INGKIIKNSEKDEIIKSNLRY(40)) and 20633 ((81)DKELTMSNVKNVSQTNFKSLY(100)) showed specific erythrocyte binding activity and inhibited merozoite invasion of erythrocytes in vitro. A monkey serum prepared against LSA-1 20630 peptide analog (CGINGKNIKNAEKPMIIKSNLRGC) inhibited merozoite invasion in vitro. The data suggest LSA-1 "High Activity Binding Peptides" could play a possible role in hepatic cell invasion as well as merozoite invasion of erythrocytes. PMID- 12895650 TI - Cicerarin, a novel antifungal peptide from the green chickpea. AB - A peptide designated cicerarin, with an N-terminal amino acid sequence VKSTGRADDDLAVKTKYLPP dissimilar from known proteins and peptides and a molecular mass of 8kDa, was isolated from seeds of the green chickpea Cicer arietinum cv green chickpea. Cicerarin was isolated with a procedure that involved ion exchange chromatography on DEAE-cellulose, affinity chromatography on Affi-gel blue gel, and gel filtration by fast protein liquid chromatography on Superdex 75. Cicerarin was unadsorbed on DEAE-cellulose and adsorbed on Affi-gel blue gel in 10mM Tris-HCl buffer (pH 7.3). Cicerarin exerted antifungal activity against Botrytis cinerea, Mycosphaerella arachidicola, and Physalospora piricola. The antifungal activity was preserved after exposure to 100 degrees C for 15min. PMID- 12895651 TI - Prediction of beta-turns with learning machines. AB - The support vector machine approach was introduced to predict the beta-turns in proteins. The overall self-consistency rate by the re-substitution test for the training or learning dataset reached 100%. Both the training dataset and independent testing dataset were taken from Chou [J. Pept. Res. 49 (1997) 120]. The success prediction rates by the jackknife test for the beta-turn subset of 455 tetrapeptides and non-beta-turn subset of 3807 tetrapeptides in the training dataset were 58.1 and 98.4%, respectively. The success rates with the independent dataset test for the beta-turn subset of 110 tetrapeptides and non-beta-turn subset of 30,231 tetrapeptides were 69.1 and 97.3%, respectively. The results obtained from this study support the conclusion that the residue-coupled effect along a tetrapeptide is important for the formation of a beta-turn. PMID- 12895652 TI - Hypophysiotropic activity of histone H3 in vitro. AB - To assess the effect of histone H3 on pituitary hormone secretion, rat anterior pituitary (AP) cells were used and growth hormone, prolactin, thyrotropin, luteinizing hormone and follicle stimulating hormone measured by radioimmunoassay. Incubation of cells with H3 (1, 6, and 30 microM) stimulated the release of all five hormones in a dose-dependent manner. This effect was blocked by preincubation of H3 with an anti-H3 antibody. Incubation of AP cells with 6 microM H3 in the presence of specific AP hormone secretagogues (GRP-6, thyrotropin-releasing hormone (TRH), gonadotropin-releasing hormone (GnRH)) showed additive effects on hormone secretion. Pharmacological experiments suggested that calcium- and diacylglycerol- (DAG) associated pathways, but not cAMP, participate in the hypophysiotropic activity of H3. Our results confirm previous evidence that histones may act as hypophysiotropic signals. PMID- 12895653 TI - Humanin peptides block calcium influx of rat hippocampal neurons by altering fibrogenesis of Abeta(1-40). AB - Humanin peptides (including HN, HNG and other mutants) were reported previously that antagonize neurotoxicity caused by various familial Alzheimer's disease (FAD) genes and Abeta derivatives. Herein, we describe the aggregation dynamics and the representative morphological characteristics of Abeta(1-40) after different time of addition humanin peptides, which revealed that (a) the interactions of both HN and HNG with Abeta(1-40) induced quick and significant increase of light-scattering intensity, and (b) HNG also caused obvious morphological alteration from fibrillary to amorphous. In the meantime, the experiments also revealed that the interaction of HNG with Abeta(1-40) could decrease Abeta(1-40)-induced calcium rise, an initial event accompanying Abeta(1 40)-induced apoptosis of cultured neurons. Our results indicate that HNG can protect neurons by altering Abeta(1-40) morphology. PMID- 12895654 TI - Co-existence of leptin- and orexin-receptors in feeding-regulating neurons in the hypothalamic arcuate nucleus-a triple labeling study. AB - The arcuate nucleus (ARC) of the hypothalamus has been identified as a prime feeding regulating center in the brain. Several feeding regulating peptides, such as neuropeptide Y (NPY) and proopiomelanocortin (POMC), are present in neurons of the ARC, which also serves as a primary targeting site for leptin, a feeding inhibiting hormone secreted predominantly by adipose tissues, and for orexin (OX) containing neurons. OX is expressed exclusively around the lateral hypothalamus, an area also established as a feeding regulating center. Some recent physiological analyses have shown that NPY- and POMC-containing neurons are activated or inactivated by leptin and OX. Moreover, we have already shown, using double immunohistochemical staining techniques, that NPY- and POMC-containing neurons express leptin receptors (LR) and orexin type 1 receptors (OX-1R). However, no morphological study has yet described the possibility of whether or not these arcuate neurons are influenced by both leptin and OX simultaneously. In order to address this issue, we performed histochemistry on ARC neurons using a triple immunofluorescence method. We found that 77 out of 213 NPY- and 99 out of 165 POMC-immunoreactive neurons co-localized with both LR- and OX-1R immunoreactivities. These findings strongly suggest that both NPY- and POMC containing neurons are regulated simultaneously by both leptin and OX. PMID- 12895655 TI - Leukocyte motility in response to neuropeptides is heparan sulfate proteoglycan dependent. AB - Activation of neuropeptide receptors on leukocytes induces chemotaxis. We determined in Boyden chambers with micropore filters, whether in human monocytes and lymphocytes this migratory response is heparan sulfate proteoglycan (HSPG) dependent. Chemotaxis toward calcitonin gene-related peptide, secretoneurin, vasoactive intestinal peptide (VIP), and substance P (SP) was abolished by removal of heparan sulfate side chains from cell surface proteoglycans or by addition of anti-syndecan-4 antibodies. Inhibition of neuropeptide-induced chemotaxis by dimethyl sphingosine (DMS), an inhibitor of sphingosine kinase, indicates transactivation of the sphingosine-1-phosphate chemotaxis pathway which was previously identified as being syndecan-4-related. Data suggest that HSPGs are involved in neuropeptide-induced chemotaxis of leukocytes. PMID- 12895656 TI - Effects of alpha-melanotropin C-terminal tripeptide analogues on macrophage NO production. AB - The C-terminal tripeptide of melanocyte-stimulating hormone, MSH (11-13) (Lys-Pro Val), possesses strong anti-inflammatory actions, which are mediated via mechanisms that are not fully understood. To shed more light into these mechanisms we have here synthesised and evaluated the activities of L- and D-Val substituted cyclic modifications of MSH (11-13) on nitric oxide (NO) in macrophage RAW 264.7 cells, as well as on binding to melanocortin receptors (MCRs) in B16-F1 and MCR expressing insect cells, and for effects on cAMP. MSH (11-13) and its analogues did neither bind to MCRs nor stimulate cAMP in RAW 264.7 and B16-F1 cells, except H-, which showed a tendency to increase cAMP at high (10-100 microM) concentrations. However, all investigated peptides dose dependently inhibited NO in LPS/IFN-gamma-stimulated RAW 264.7, cells with a structure activity relationship suggesting the existence of a distinct receptive site. This site appears to be distinct from the MCRs and not linked with cAMP. PMID- 12895657 TI - Regulation of TNF-alpha secretion by a specific melanocortin-1 receptor peptide agonist. AB - The lack of specific pharmacological tools has impeded the evaluation of the role of each melanocortin receptor (MCR) subtype in the myriad physiological effects of melanocortins. 154N-5 is an octapeptide (MFRdWFKPV-NH(2)) that was first identified as an MC1R antagonist in Xenopus melanophores [J. Biol. Chem. 269 (1994) 29846]. In this manuscript, we show that 154N-5 is a specific agonist for human and murine MC1R. The peptide has negligible activity at MC3R and MC4R and is 25-fold less potent and a weak agonist at MC5R. 154N-5 was tested in both a cellular and an animal model of tumor necrosis factor-alpha (TNF-alpha) secretion. The inhibitory efficacy of 154N-5 on TNF-alpha secretion in both models was similar to the nonselective agonist NDP-alpha-melanocyte stimulating hormone (NDP-alphaMSH), thus, we conclude that inhibition of TNF-alpha secretion by melanocortin peptides is mediated by MC1R. 154N-5 is a valuable new tool for the evaluation of specific contribution of MC1R agonism to physiological and pathological processes. PMID- 12895658 TI - Regulation of ACTH levels in anterior pituitary cells during stimulated secretion: evidence for aspartyl and cysteine proteases in the cellular metabolism of ACTH. AB - The regulation of cellular levels of adrenocorticotropin hormone (ACTH) in response to stimulated secretion was investigated to define the extent of cellular depletion of ACTH and subsequent increases to replenish ACTH levels in anterior pituitary cells (in primary culture). Treatment of cells with secretagogues for short-term incubation times (hours) resulted in extensive depletion of cellular ACTH. Corticotropin releasing factor (CRF) induced depletion of cellular levels of ACTH by 60-70% of control levels. The CRF-induced reduction of cellular ACTH was inhibited by the glucocorticoid dexamethasone. Phorbol myristate acetate (PMA), which stimulates protein kinase C (PKC), reduced ACTH levels by 50-60%. Forskolin, a stimulator of cAMP production, produced a moderate reduction in cellular ACTH. During prolonged incubation of cells (2 days) with these secretagogues, further reduction of ACTH levels by 70-80% was observed. However, increased cellular levels of ACTH occurred with continued treatment of cells with secretagogues, which provided nearly complete replenishment of cellular ACTH after 5 days treatment with secretagogues. Notably, the rising levels of cellular ACTH were inhibited by the aspartyl protease inhibitor acetyl-pepstatin A, and by the cysteine protease inhibitor E64d. These results demonstrate that depletion and recovery of ACTH levels are coordinately regulated, and that the increases in cellular levels of ACTH during the recovery phase involves participation of aspartyl and cysteine proteases. Thus, aspartyl and cysteine proteases may be involved in the cellular metabolism of ACTH. PMID- 12895659 TI - Stimulation of rat pancreatic exocrine secretion by urocortin and corticotropin releasing factor. AB - Neural and hormonal mechanisms control pancreatic secretion. The effects of the corticotropin releasing factor (CRF) related neuropeptide urocortin (UCN) on pancreatic exocrine secretion were examined. In anesthetized male rats, pancreatic secretion volume and total protein were assayed. UCN increased pancreatic secretory volume and protein secretion and potentiated cholecytokinin stimulated protein secretion. Astressin, a non-specific CRF receptor antagonist, inhibited UCN-stimulated protein output while CRF(2) receptor antagonist, antisauvagine-30, was without effect. Atropine, but not subdiaphragmatic vagotomy, inhibited UCN-mediated secretion. In acinar cells, UCN did not stimulate release of amylase nor intracellular cAMP. UCN is a pancreatic exocrine secreatagogue with effects mediated through cholinergic intrapancreatic neurons. PMID- 12895660 TI - Anti-analgesic activity of enterostatin (VPDPR) is mediated by corticosterone. AB - Although enterostatin (VPDPR) inhibited morphine-induced analgesia, it had no affinity for mu-opioid receptors. VPDPR administration was reported to elevate serum corticosterone levels. We found that corticosterone exhibited a similar anti-analgesic effect selective for mu-opioid. Furthermore, the anti-analgesic effect of VPDPR was inhibited by RU486, an antagonist for the glucocorticoid receptor. The anti-analgesic effect of VPDPR was not observed in adrenalectomized mice. These results suggest that the anti-analgesic activity of VPDPR is mediated by corticosterone released from the adrenal cortex. PMID- 12895661 TI - Endomorphin-1, an endogenous mu-opioid receptor agonist, improves apomorphine induced impairment of prepulse inhibition in mice. AB - The present study was designed to examine the effects of the endogenous mu-opioid receptor agonist endomorphin-1 on prepulse inhibition (PPI) in mice. Although apomorphine (1mg/kg) produced a marked decrease in PPI, endomorphin-1 (17.5 microg) had no marked effects on PPI or startle amplitude in normal mice. Endomorphin-1 (17.5 microg) inhibited the apomorphine (1mg/kg)-induced decrease in PPI. beta-Funaltrexamine (5 microg), a mu-opioid receptor antagonist, did not significantly antagonize the effects of endomorphin-1 (17.5 microg). Naloxonazine (35 mg/kg), a mu(1)-opioid receptor antagonist, antagonized the effects of endomorphin-1 (17.5 microg) on the apomorphine (1mg/kg)-induced decrease in PPI, whereas naloxonazine (35 mg/kg) itself was without significant effects on the apomorphine (1mg/kg)-induced decrease. These results suggest that endomorphin-1 alleviates the impairment of PPI resulting from the hyperactivity of dopaminergic neurotransmission through the mediation of mu(1)-opioid receptors. PMID- 12895662 TI - Cardiovascular responses to central administration of mu and kappa opioid receptor agonist and antagonist in normal rats. AB - The response to centrally administered beta-endorphin has been characterized by decreasing sympathetic nervous activity and decreased cardiovascular tone. We investigated the effect of the central administration of both mu and kappa opioid receptor agonist and antagonists on cardiovascular responses. The administration of the mu agonist, DAMGO (0.2nmol) increased the mean arterial pressure (MAP) and stimulated iliac vasoconstriction while higher doses (2 and 20nmol) decreased MAP and stimulated iliac vasodilation. The administration of the kappa receptor agonist, Dynorphin decreased the MAP and stimulated superior mesenteric vasodilation. beta-Funaltrexamine reduced MAP and superior mesenteric vasodilation while nor-binaltorphimine increased MAP and iliac and superior mesenteric vasoconstriction. We conclude that mu receptor activation decrease or increase MAP depending on the mu agonist concentration. However, kappa receptor activation is consistently associated with a decrease in MAP. PMID- 12895663 TI - Angiotensinase activities in the kidney of renovascular hypertensive rats. AB - In spite of the well-known contribution of angiotensin II (Ang II) in the pathogenesis of Goldblatt two-kidney one clip (G2K1C) hypertension, the importance of other Ang peptides, such as Ang III, Ang IV or Ang 2-10, is scarcely understood. The functional status of these peptides depends on the action of several aminopeptidases called angiotensinases. The metabolism of Ang III to Ang IV by aminopeptidase M (AlaAP) and of Ang I to Ang 2-10 by aspartyl aminopeptidase (AspAP) was evaluated in the renal cortex and medulla of normotensive (Sham-operated) and hypertensive (G2K1C) rats, treated or not with the AT(1) receptor antagonist valsartan. The results demonstrated a highly significant increase of membrane-bound (MEMB) AlaAP in the cortex of the non ischemic kidney of G2K1C rats compared with the kidney of normal rats and with the clipped kidney of G2K1C rats. This suggests an increased formation of Ang IV in the non-clipped kidney of G2R1C rats. Valsartan reduced MEMB AlaAP and AspAP activities in the renal cortex of normotensive and in the clipped kidney of hypertensive rats. The reduced metabolism of Ang III may prolong its half-life in valsartan-treated animals. These results suggest a role for AlaAP in renovascular hypertension. In addition, the higher AspAP activity of the renal cortex compared to medulla reflects its relative functional difference between both locations. PMID- 12895664 TI - Substance P in the uterine cervix, dorsal root ganglia and spinal cord during pregnancy and the effect of estrogen on SP synthesis. AB - Prior to parturition the non-pliable uterine cervix undergoes a ripening process ("softens" and dilates) to allow a timely passage of the fetus at term. The exact mechanism(s) triggering and involved in cervical ripening are unknown, though evidence for a role for sensory neurons and their contained neuropeptides is emerging. Moreover, an apparent increase in neuropeptide immunoreactive nerves occurs in the cervix during pregnancy, maternal serum estrogen levels rise at term and uterine cervix-related L6-S1 dorsal root ganglia (DRG) sensory neurons express estrogen receptor (ER) and neuropeptides. Thus, we sought to test the hypothesis that the neuropeptide substance P (SP) changes biosynthesis and release over pregnancy, that estrogen, acting via the ER pathway, increases synthesis of SP in DRG, and that SP is utilized in cervical ripening at late pregnancy. Using immunohistochemistry, in situ hybridization, reverse transcriptase-polymerase chain reaction (RT-PCR) and radioimmunoassay (RIA), we investigated coexpression of ER-alpha/beta and SP; differential expression of ER alpha and -beta mRNA in DRG neurons; SP synthesis in DRG; and changes in SP concentration in the cervix, DRG and spinal cord over pregnancy. In addition, the effect of exogenous estrogen on SP synthesis in L6-S1 DRG of ovariectomized rats was examined. SP-immunoreactive neurons expressed ER-alpha and ER-beta. SP synthesis (expressed as beta-PPT mRNA label) was prominent in small DRG neurons. SP concentration increased in the L6-S1 DRG and spinal cord segments, with a peak at Day 20 of gestation, but decreased in the cervix during the first two trimesters, with a rise over the last trimester to Day 10 levels. SP and ER-alpha mRNA synthesis increased in DRG over pregnancy but ER-beta mRNA levels were largely unchanged. When ovariectomized rats were treated with exogenous estrogen, SP mRNA synthesis in the DRG increased in a dose-related manner, an effect blocked by ER blocker ICI 182 780. From these results, we postulate that synthesis of SP in L6-S1 DRG and utilization in the cervix increase over pregnancy and this synthesis is under influence of the estrogen-ER system, most likely ER-alpha. We postulate that SP may play a role in cervical ripening and, consequently in the birth process. PMID- 12895665 TI - Inhibition of recombinant dipeptidyl peptidase III by synthetic hemorphin-like peptides. AB - In order to find the most effective antagonist for dipeptidyl peptidase III degrading enkephalin, we synthesized hemorphin-like pentapeptides with aliphatic or aromatic amino acids at the N-termini, such as VVYPW, LVYPW, IVYPW, YVYPW, FVYPW and WVYPW. Among those pentapeptides, IVYPW and WVYPW showed the strongest inhibitory activity toward rDPP III. The K(i) values of IVYPW and WVYPW were 0.100+/-0.011 and 0.126+/-0.015 microM (mean+/-S.E.), respectively. The order of K(i) values was Ile> or =Trp>Phe> or =Tyr>Leu>Ala>Val>Ser>Gly. rDPP III activity is inhibited in a non-competitive manner by these peptides. The peptide VYPW did not inhibit rDPP III activity, but the sequence is essential for the expression of inhibitory activity. PMID- 12895666 TI - Emotion regulation and touch in infants: the role of cholecystokinin and opioids. AB - Behavioral-pharmacological research in infant rats supports the role of cholecystokinin (CCK) and opioid peptides in mediating early learning of new associations with aspects of the nest and dam, such as maternal odor, milk, and contact. The current paper reviews research that examines the hypothesis that these neuropeptide systems are further involved in mediating emotion regulation in infants, thus playing a role in the emergence of stress-reactivity and other motivational systems. The beneficial effects of maternal proximity, handling, and touch on the development of emotion regulation have been demonstrated in both human and animal models. Interventions that promote tactile stimulation of the infant ("touch therapy") and infant-mother contact ("skin-to-skin contact" or "kangaroo care") have been shown to improve the infant's ability to self regulate, and to moderate the effects of some risk factors. Theoretical perspectives and empirical findings regarding emotion regulation in infants are first discussed. This is followed by a review of work providing evidence in animal models (and suggestive evidence in humans) for the importance of CCK and opioid neuropeptides in affecting infant emotion regulation and the impact of touch-based interventions, in particular in the context of infant-mother attraction, contact, separation, and attachment. PMID- 12895668 TI - Plants and the central nervous system. AB - This review article draws the attention to the many species of plants possessing activity on the central nervous system (CNS). In fact, they cover the whole spectrum of central activity such as psychoanaleptic, psycholeptic and psychodysleptic effects, and several of these plants are currently used in therapeutics to treat human ailments. Among the psychoanaleptic (stimulant) plants, those utilized by human beings to reduce body weight [Ephedra spp. (Ma Huang), Paullinia spp. (guarana), Catha edulis Forssk. (khat)] and plants used to improve general health conditions (plant adaptogens) were scrutinized. Many species of hallucinogenic (psychodysleptic) plants are used by humans throughout the world to achieve states of mind distortions; among those, a few have been used for therapeutic purposes, such as Cannabis sativa L., Tabernanthe iboga Baill. and the mixture of Psychotria viridis Ruiz and Pav. and Banisteriopsis caapi (Spruce ex Griseb.) C.V. Morton. Plants showing central psycholeptic activities, such as analgesic or anxiolytic actions (Passiflora incarnata L., Valeriana spp. and Piper methysticum G. Forst.), were also analysed.Finally, the use of crude or semipurified extracts of such plants instead of the active substances seemingly responsible for their therapeutic effect is discussed. PMID- 12895669 TI - Plants used in Chinese and Indian traditional medicine for improvement of memory and cognitive function. AB - In traditional practices of Ayurvedic and Chinese medicine, numerous plants have been used to treat cognitive disorders, including neurodegenerative diseases such as Alzheimer's disease (AD). An ethnopharmacological approach has provided leads to identifying potential new drugs from plant sources, including those for cognitive disorders. Many drugs currently available in Western medicine were originally isolated from plants, or are derived from templates of compounds isolated from plants. Some anticholinesterase (anti-ChE) alkaloids isolated from plants have been investigated for their potential in the treatment of AD, and are now in clinical use. Galantamine, isolated from several plants including Lycoris radiata Herb., which was used in traditional Chinese medicine (TCM), is licensed in the United Kingdom for the treatment of mild to moderate AD. Various other plant species have shown pharmacological activities relevant to the treatment of cognitive disorders, indicating potential for therapeutic use in disorders such as AD. This article reviews some of the plants and their active constituents that have been used in traditional Ayurvedic medicine and TCM for their reputed cognitive-enhancing or antiageing effects. Plants and their constituents with pharmacological activities that may be relevant for the treatment of cognitive disorders, including enhancement of cholinergic function in the central nervous system (CNS), anti-inflammatory and antioxidant activities, are discussed. PMID- 12895670 TI - Clitoria ternatea and the CNS. AB - The present investigation was aimed at determining the spectrum of activity of the methanolic extract of Clitoria ternatea (CT) on the CNS. The CT was studied for its effect on cognitive behavior, anxiety, depression, stress and convulsions induced by pentylenetetrazol (PTZ) and maximum electroshock (MES). To explain these effects, the effect of CT was also studied on behavior mediated by dopamine (DA), noradrenaline, serotonin and acetylcholine. The extract decreased time required to occupy the central platform (transfer latency, TL) in the elevated plus maze (EPM) and increased discrimination index in the object recognition test, indicating nootropic activity. The extract was more active in the object recognition test than in the EPM. The extract increased occupancy in the open arm of EPM by 160% and in the lit box of the light/dark exploration test by 157%, indicating its anxiolytic activity. It decreased the duration of immobility in tail suspension test (suggesting its antidepressant activity), reduced stress induced ulcers and reduced the convulsing action of PTZ and MES. The extract exhibited tendency to reduce the intensity of behavior mediated via serotonin and acetylcholine. The effect on DA- and noradrenaline-mediated behavior was not significant. In conclusion, the extract was found to possess nootropic, anxiolytic, antidepressant, anticonvulsant and antistress activity. Further studies are necessary to isolate the active principle responsible for the activities and to understand its mode of action. PMID- 12895671 TI - 6-methylapigenin and hesperidin: new valeriana flavonoids with activity on the CNS. AB - Valerian is an ancient tranquillizing drug obtained from the underground organs of several Valeriana species. Its active principles were assumed to be terpenoids in the form of valepotriates and/or as components of the essential oil. However, unknown active compounds were not discarded and synergic effects were suspected. We have recently isolated 6-methylapigenin (MA) from Valeriana wallichii and proved that it is a benzodiazepine binding site (BDZ-bs) ligand [Planta Med. 68 (2002) 934]. The present paper is the first report of the presence of 2S(-) hesperidin in valeriana and describes that it has sedative and sleep-enhancing properties. MA, in turn, was found to have anxiolytic properties and was able to potentiate the sleep-enhancing properties of hesperidin (HN).MA and HN are new members of the growing family of natural flavonoids with activity on the CNS, and their properties suggest that they are promising drug leads in the field. PMID- 12895672 TI - Adaptogenic activity of Withania somnifera: an experimental study using a rat model of chronic stress. AB - Withania somnifera (WS) Dunal is classified in Ayurveda, the ancient Hindu system of medicine, as a rasayana, a group of plant-derived drugs reputed to promote physical and mental health, augment resistance of the body against disease and diverse adverse environmental factors, revitalise the body in debilitated conditions and increase longevity. These attributes are remarkably similar to the properties ascribed to adaptogens like Panax ginseng (PG) in contemporary medicine. As such, the adaptogenic activity of a standardised extract of WS roots was investigated against a rat model of chronic stress (CS). The stress procedure was mild, unpredictable footshock, administered once daily for 21 days to adult male Wistar rats. CS induced significant hyperglycaemia, glucose intolerance, increase in plasma corticosterone levels, gastric ulcerations, male sexual dysfunction, cognitive deficits, immunosuppression and mental depression. These CS induced perturbations were attenuated by WS (25 and 50 mg/kg po) and by PG (100 mg/kg po), administered 1 h before footshock for 21 days. The results indicate that WS, like PG, has significant antistress adaptogenic activity, confirming the clinical use of the plant in Ayurveda. PMID- 12895673 TI - Long-term effects of an Apocynum venetum extract on brain monoamine levels and beta-AR density in rats. AB - The present study was designed to get further insight into the mode of antidepressant action of an extract prepared of the leaves of Apocynum venetum L. (AV). To evaluate biochemical changes, we used a high-performance liquid chromatography system to examine the effects of short-term (2 weeks) and long term (8 weeks) administration of imipramine (15 mg/kg po) and an AV-extract (15, 60 and 250 mg/kg) on regional levels of serotonin (5-HT), norepinephrine (NE), dopamine (DA) and their metabolites in the rat hypothalamus, striatum and hippocampus. Pronounced changes in 5-HT, NE and DA levels were detected mainly after 8 weeks of daily imipramine treatment. Similar to imipramine, AV-extract reduced NE and DA concentrations after 8 weeks, whereas it failed to affect 5-HT levels. We speculate that the decrease in NE levels after chronic AV treatment might be based partly on the subsensitivity of presynaptic alpha(2)-receptors. In addition to the determination of central monoamine concentrations, quantitative radioligand receptor-binding studies were used to examine the effects of long term administration of imipramine and AV-extract on beta-adrenergic binding in rat frontal cortex. [125I]CYP binding to beta-adrenergic receptors was found to be decreased after 8 weeks treatment with imipramine, whereas AV-extract had no effect on beta-receptor binding. PMID- 12895674 TI - Perinatal exposure to delta 9-tetrahydrocannabinol increases presynaptic dopamine D2 receptor sensitivity: a behavioral study in rats. AB - The endogenous cannabinoid system is a relevant modulator of dopaminergic synapses in dorsal striatum. Perinatal exposure to cannabinoid receptor agonists has been described to affect the development of dopaminergic circuits in rat brain. The epigenetic alterations described affected both dopamine neurons and dopamine receptor-expressing neurons. The present work has been designed to explore the effects of maternal exposure to orally delivered Delta(9) tetrahydrocannabinol, (Delta(9)-THC 0.1, 0.5, 2 mg/kg) on the behavioural responses to the dopamine receptor agonists apomorphine (0.1 mg/kg) and quinpirole (0.5 mg/kg), at doses that target presynaptic dopamine D2 receptors. Maternal exposure to Delta(9)-THC affected both the developmental pattern of motor behaviours, and the behavioural responses to acute injections of apomorphine and quinpirole, tested in an open field. The effects were sex dimorphic, being more intense in male animals. Perinatal exposure to Delta(9)-THC resulted in enhanced presynaptic dopamine D2 receptor mediated responses such as immobility and inhibition of locomotion. Additionally, postsynaptic dopamine D2 receptor agonist-induced stereotypes were reduced in the group exposed to the highest dose of Delta(9)-THC (2 mg/kg). However, the late-onset pattern of behavioural activation observed after acute quinpirole exposure was equal in vehicle- and cannabinoid-treated animals. These effects suggest that perinatal exposure to Delta(9)-THC affects the functionality of dopaminergic autoreceptors, inducing a greater sensitivity to the presynaptic actions of dopamine D(2) receptor agonists. PMID- 12895675 TI - Effects of perinatal exposure to delta 9-tetrahydrocannabinol on operant morphine reinforced behavior. AB - The present study examined the effects of Delta(9)-tetrahydrocannabinol (Delta(9) THC) when administered during the perinatal period on morphine self administration in adulthood. To this end, pregnant Wistar rats were daily exposed to Delta(9)-THC from the fifth day of gestation up to pup weaning, when they were separated by gender and left to mature to be used for analyses of operant food- and morphine-reinforced behavior in a progressive ratio (PR) schedule. We also analyzed dopaminergic activity (DOPAC/DA) in reward-related structures during specific phases of the behavioral study. In both reinforcement paradigms, food and morphine, females always reached higher patterns of self-administration than males, but this occurred for the two treatment groups, Delta(9)-THC or vehicle. These higher patterns measured in females corresponded with a higher DOPAC/DA in the nucleus accumbens prior to the onset of morphine self-administration in comparison to males. Interestingly, DOPAC/DA was lower in Delta(9)-THC-exposed females compared to oil-exposed females and similar to oil- and Delta(9)-THC exposed males. In addition, Delta(9)-THC-exposed females also exhibited a reduction in DOPAC/DA in the ventral tegmental area, which did not exist in males. All these changes, however, disappeared after 15 days of morphine self administration and they did not reappear after 15 additional days of extinction of this response. Our data suggest that females are more vulnerable than males in a PR schedule for operant food and morphine self-administration; perinatal Delta(9)-THC exposure is not a factor influencing this vulnerability. The neurochemical analysis revealed that the activity of limbic dopaminergic neurons prior to morphine self-administration was higher in females than males, as well as that the perinatal Delta(9)-THC treatment reduced the activity of these neurons only in females, although this had no influence on morphine vulnerability in these animals. PMID- 12895676 TI - Effects of the synthetic cannabinoid nabilone on spatial learning and hippocampal neurotransmission. AB - Cannabinoids, the active components of marijuana, affect memory and hippocampal neurotransmission. It has been claimed that nabilone, a synthetic cannabinoid endowed with antiemetic properties, has a peculiar profile of actions. We studied the effects of the drug on spatial learning and in vitro hippocampal CA1 electrophysiology in the rat. Nabilone (0.1, 0.5, and 1.0 mg/kg ip) does not impair place learning in a water maze task, whereas Delta(8)-tetrahydrocannabinol (Delta(8)-THC) disrupts this function. At concentrations ranging from 1 nM to 10 microM nabilone does not influence basal glutamatergic neurotransmission, which is decreased by Delta(8)-THC. Although cannabinoids have been consistently reported to affect synaptic plasticity, nabilone 1 microM does not change paired pulse facilitation, long-term potentiation and the magnitude of long-term depression. However, the time course of the latter phenomenon is significantly changed by the drug, the depression being lower than in control experiments from 7 to 35 min postinduction. Altogether, our data indicate that there might be differences in the effects of agonists for central cannabinoid receptors, which could help to understand the pharmacology of this class of molecules. The results also suggest that amnesia induced by cannabinoids be possibly related to their effects on hippocampal neurotransmission. The study supports the use of nabilone in conditions the course of which is complicated by cognitive impairment. PMID- 12895677 TI - Plant derivatives in the treatment of alcohol dependency. AB - The present review summarizes the findings of the effects of extracts of purified compounds from several plants on alcohol intake in alcohol-preferring rats. These include St. John's wort (Hypericum perforatum, HPE), kudzu (Pueraria lobata) and ibogaine (Tabernanthe iboga). Alcohol-preferring (P), Marchigian Sardinian (msP), high-alcohol-drinking (HAD), Fawn-Hooded (FH) rats were allowed to drink alcohol or water voluntarily to establish baseline levels. Pure compounds (puerarin, daidzin, daidzein or analogs) isolated from kudzu, extracts from HPE or ibogaine and its analog were given by either intraperitoneal or oral administration. After acute administration, all agents dose-dependently reduced alcohol intake with minimal effects on food intake. Puerarin and HPE were also effective following chronic treatment. Overall, it is clear that pure compounds (daidzin, puerarin), extracts from St. John's wort, ibogaine and an ibogaine analog suppress alcohol intake in animal models of excessive drinking with minimal effects on other appetitive behaviors. Although the true mechanisms of action of these compounds on alcohol intake are not fully understood, with the current information, it appears that these compounds exert their effects by modulating several neuronal systems implicated in drinking behavior. However, their role in the future of pharmacotherapy for alcoholism will depend upon the outcome of carefully conducted clinical trials. PMID- 12895678 TI - Anti-addictive actions of an iboga alkaloid congener: a novel mechanism for a novel treatment. AB - 18-Methoxycoronaridine (18-MC), a novel iboga alkaloid congener that decreases drug self-administration in several animal models, may be a potential treatment for multiple forms of drug abuse. In animal models, 18-MC reduced intravenous morphine, cocaine, methamphetamine and nicotine self-administration, oral alcohol and nicotine intake, and attenuated signs of opioid withdrawal, but had no effect on responding for a nondrug reinforcer (water) and produced no apparent toxicity [Brain Res. 719 (1996) 29; NeuroReport 11 (2000) 2013; Pharmacol. Biochem. Behav. 58 (1997) 615; Psychopharmacology (Berl.) 139 (1998) 274; NeuroReport 9 (1998) 1283; Ann. N. Y. Acad. Sci. 914 (2000) 369]. Consistent with a relationship among drug sensitization, mesolimbic dopamine, and drug-seeking behavior, 18-MC also blocked the sensitized dopamine responses to morphine and cocaine in the nucleus accumbens. An extensive series of receptor studies showed that 18-MC was most potent and somewhat selective as an antagonist at alpha3beta4 nicotinic receptors. Low-dose combinations of 18-MC with other drugs known to have this same action (e.g., mecamylamine, dextromethorphan, bupropion) decreased morphine, methamphetamine, and nicotine self-administration in rats at doses that were ineffective if administered alone. Together, the data support the hypothesis that diencephalic pathways having high densities of alpha3beta4 nicotinic receptors modulate mesocorticolimbic pathways more directly involved in drug reinforcement. Antagonists of alpha3beta4 nicotinic receptors may represent a totally novel approach to treating multiple addictive disorders, and 18-MC might be the first of a new class of synthetic agents acting via this novel mechanism and having a broad spectrum of activity. PMID- 12895680 TI - Activities of extract and constituents of Banisteriopsis caapi relevant to parkinsonism. AB - Dopamine deficiency is characteristic of Parkinson's disease (PD) and treatments aim at elevating levels by administration of its precursor L dihydroxyphenylalanine (L-DOPA), or inhibiting monoamine oxidases (MAOs), thus preventing its breakdown. Reports of improvements in PD patients treated with Banisteriopsis caapi extracts stimulated investigation of B. caapi stem extract and its two ingredients, harmine and harmaline for these activities. Tests for MAO inhibition using liver homogenate showed that extract and harmaline showed a concentration-dependent inhibition of MAO A (IC(50) 1.24 microg/ml and IC(50) 4.54 nM, respectively) but had little effect on MAO B activity. The extract at 2.5 mg/ml caused a highly significant increase in release of [3H]dopamine from rat striatal slices, as did 200 microM harmine and 6 microM harmaline. In both these experiments, the amount of harmine present could not account for the total activity of the extract. The ability of harmine and harmaline to stimulate dopamine release is a novel finding. These results give some basis to the reputed usefulness of B. caapi stem extract in the treatment of PD. PMID- 12895679 TI - NPI-031G (puerarin) reduces anxiogenic effects of alcohol withdrawal or benzodiazepine inverse or 5-HT2C agonists. AB - Because extracts of kudzu have been used as a hangover remedy in China for many centuries, we tested the ability of NPI-031G (puerarin), an isoflavone isolated from kudzu, to counteract anxiogenic effects associated with withdrawal from chronic alcohol exposure. NPI-O31G (50 and 150 mg/kg ip) significantly increased the social interaction and locomotor activity reduced by withdrawal from 17 days of alcohol (7%) diet. The effects of NPI-031G resembled those of the benzodiazepine antagonist, flumazenil (5 mg/kg), and the 5-HT(2C) antagonist, SB 242084 (1 mg/kg). In a separate study, control rats were pretreated with NPI-031G (30 min) and then given the anxiogenic compounds DMCM, a benzodiazepine inverse agonist, or Ro 600175, a 5-HT(2C) agonist. NPI-031G significantly counteracted the reduction in social interaction induced by either compound. To identify a potential mechanism of action of NPI-031G, synaptoneurosomes were isolated from the cerebral cortex of untreated rats and chloride uptake assays were carried out. NPI-031G did not have any effect on the stimulation of chloride uptake by muscimol, a GABA(A) agonist. However, it reduced the potentiation of muscimol stimulated chloride uptake by flunitrazepam, a benzodiazepine agonist, at a concentration of 100 microM. A reduction in [3H]flunitrazepam binding was also seen at this concentration. These findings are consistent with NPI-031G being a weak benzodiazepine site antagonist. PMID- 12895681 TI - Pharmacological evidence for antidementia effect of Choto-san (Gouteng-san), a traditional Kampo medicine. AB - To clarify the clinical efficacy of one of the traditional medicines in the treatment of patients with vascular dementia, we investigated the pharmacological activities of Choto-san in animal models. Pretreatment with Choto-san (0.75-6.0 g/kg po), a component herb, Chotoko (75-600 mg/kg po), and indole alkaloids and phenolic fractions of Chotoko prevented ischemia-induced impairment of spatial learning behaviour in water maze performance of mice. A single administration of Choto-san (0.5 to 6.0 g/kg po) or Chotoko (Uncaria genus) produced a dose dependent antihypertensive effect in spontaneously hypertensive rats (SHR) and partly inhibited the induction of the apoplexy in stroke-prone SHR (SHR-SP). Choto-san, Chotoko, and its phenolic constituents, (-)epicatechin and caffeic acid, significantly protected NG108-15 cells from injury induced by H(2)O(2) exposure in vitro and also inhibited lipid peroxidation in the brain homogenate. Indole alkaloids, rhynchophylline and isorhynchophylline (1-100 microM), reversibly reduced N-methyl-D-aspartate (NMDA)-induced current concentration dependently in NMDA receptor-expressed Xenopus oocytes. These results suggest that antidementia effects of Choto-san are due to antihypertensive, free radical scavenging and antiexcitotoxic effects, which are attributed at least partly to phenolic compounds and indole alkaloids contained in Chotoko. PMID- 12895682 TI - Ptychopetalum olacoides, a traditional Amazonian "nerve tonic", possesses anticholinesterase activity. AB - The cholinergic hypothesis of Alzheimer disease (AD) has provided the rationale for the current pharmacotherapy of this disease, in an attempt to downgrade the cognitive decline caused by cholinergic deficits. Nevertheless, the search for potent and long-acting acetylcholinesterase (AChE) inhibitors that exert minimal side effects to AD patients is still an ongoing effort. Amazonian communities use traditional remedies prepared with Ptychopetalum olacoides (PO, Olacaceae) roots for treating various central nervous system conditions, including those associated with aging. The fact that PO ethanol extract (POEE) has been found to facilitate memory retrieval in the step down procedure in young and aged mice prompt us to evaluate its effects on AChE activity in memory relevant brain areas. POEE significantly inhibited AChE activity in vitro in a dose- and time dependent manner in rat frontal cortex, hippocampus and striatum; a significant inhibition was also found in these same brain areas of aged (14 months) mice after acute administration of POEE (100 mg/kg ip). We propose that such AChE inhibitory activity is a neurochemical correlate of a number of therapeutic properties traditionally claimed for P. olacoides, particularly those associated with cognition. PMID- 12895683 TI - Salvia for dementia therapy: review of pharmacological activity and pilot tolerability clinical trial. AB - S. lavandulaefolia Vahl. (Spanish sage) extracts and constituents have demonstrated anticholinesterase, antioxidant, anti-inflammatory, oestrogenic and CNS depressant (sedative) effects all of which are currently relevant to the treatment of Alzheimer's disease (AD). The essential oil inhibits the enzyme acetylcholinesterase (AChE) from human brain tissue and bovine erythrocyte and individual monoterpenoid constituents inhibit AChE with varying degrees of potency. In vivo AChE inhibition of select brain (striatal and hippocampal over cortical) AChE was obtained following oral administration of the essential oil to rats. In a study in healthy volunteers essential oil administration produced significant effects on cognition. In a pilot open-label study involving oral administration of the essential oil to patients with AD, a significant increase in diastolic and systolic blood pressure was observed in two patients, however this may have been due primarily to preexisting hypertension and there were no abnormalities in other vital signs or blood samples during the trial period. Although an open label trial is not free from practice effects or rater-caregiver expectations, statistically significant differences between baseline and 6 weeks treatment were a reduction in neuropsychiatric symptoms and an improvement in attention. PMID- 12895684 TI - Synergistic and antagonistic interactions of anticholinesterase terpenoids in Salvia lavandulaefolia essential oil. AB - In vitro anticholinesterase activities of eight commercially available terpenoid constituents of Salvia lavandulaefolia have been investigated. These included 1,8 cineole, camphor, alpha-pinene, beta-pinene, borneol, caryophyllene oxide, linalool and bornyl acetate. Dose-dependent inhibition of acetylcholinesterase (AChE) by these chemical constituents was determined using the method of Ellman [Biochem. Pharmacol. 7 (1961) 88]. The IC50 value of 1,8-cineole was 0.06+/-0.01 mg/ml similar to that of the essential oil (0.05+/-0.01 mg/ml). Analyses of the expected inhibitions based on the prediction of a zero interactive response of a combination at its naturally occurring ratios were carried out in comparison with observed inhibition. Minor synergy was apparent in 1,8-cineole/alpha-pinene and 1,8-cineole/caryophyllene oxide combinations, with interaction indexes not exceeding 0.5. In contrast, a combination of camphor and 1,8-cineole was antagonistic with an interaction index of 2. A combination of all eight compounds was zero interactive. A combination of six constituents, excluding 1,8-cineole and camphor, was used to compare the method of expected response of a combination with a method of summation. These findings reveal that the inhibitory activity of the oil results from a complex interaction between its constituents, which produce both synergistic and antagonistic responses between the component terpenes. Understanding such interactions is important in comparing species on the basis of chemical composition. PMID- 12895685 TI - Salvia lavandulaefolia (Spanish sage) enhances memory in healthy young volunteers. AB - Sage (Salvia) has a longstanding reputation in British herbal encyclopaedias as an agent that enhances memory, although there is little evidence regarding the efficacy of sage from systematized trials. Based on known pharmacokinetic and binding properties, it was hypothesised that acute administration of sage would enhance memory in young adult volunteers. Two experiments utilised a placebo controlled, double-blind, balanced, crossover methodology. In Trial 1, 20 participants received 50, 100 and 150 microl of a standardised essential oil extract of Salvia lavandulaefolia and placebo. In Trial 2, 24 participants received 25 and 50 microl of a standardised essential oil extract of S. lavandulaefolia and placebo. Doses were separated by a 7-day washout period with treatment order determined by Latin squares. Assessment was undertaken using the Cognitive Drug Research computerised test battery prior to treatment and 1, 2.5, 4 and 6 h thereafter. The primary outcome measures were immediate and delayed word recall. The 50 microl dose of Salvia essential oil significantly improved immediate word recall in both studies. These results represent the first systematic evidence that Salvia is capable of acute modulation of cognition in healthy young adults. PMID- 12895686 TI - The psychopharmacology of huperzine A: an alkaloid with cognitive enhancing and neuroprotective properties of interest in the treatment of Alzheimer's disease. AB - Huperzine A (HupA), extracted from a club moss (Huperzia serrata), is a sesquiterpene alkaloid and a powerful and reversible inhibitor of acetylcholinesterase (AChE). It has been used in China for centuries for the treatment of swelling, fever and blood disorders. It has demonstrated both memory enhancement in animal and clinical trials and neuroprotective effects. Recently it has undergone double-blind, placebo-controlled clinical trials in patients with Alzheimer's disease (AD), with significant improvements both to cognitive function and the quality of life. Most of the clinical trials are from China, but HupA and derivatives are attracting considerable interest in the West, where AD is a major and growing concern. Furthermore, both animal and human safety evaluations have demonstrated that HupA is devoid of unexpected toxicity. Other interesting aspects of HupA pharmacological profile relate to its neuroprotective properties: it has been shown in animal studies that HupA can be used as a protective agent against organophosphate (OP) intoxication and that it reduces glutamate-induced cell death. PMID- 12895687 TI - Ginseng: potential for the enhancement of cognitive performance and mood. AB - Ginseng has been used medicinally in the Far East for several millennia and is currently one of the most widely taken herbal products throughout the world. It has been attributed with a plethora of physiological effects that could potentially benefit cognitive performance or mood. Studies involving animals show that ginseng and its constituent ginsenosides can modulate indices of stress, fatigue, and learning. However, there is a lack of adequately controlled research showing behavioural effects following chronic administration to humans. Recent research has demonstrated that single doses of ginseng most notably engender cognitive benefits in terms of improved memory, but can also be associated with 'costs' in terms of attention task deficits following less mnemonically beneficial doses. A single dose of ginseng has also been shown to modulate cerebroelectrical (EEG) activity. It is suggested that ginseng would benefit from rigorous research further delineating its acute effects and exploring the relationship between acute effects and those seen during and following chronic administration regimens. PMID- 12895688 TI - Electroencephalograph effects of single doses of Ginkgo biloba and Panax ginseng in healthy young volunteers. AB - Both Ginkgo biloba and Panax ginseng exert a number of physiological effects and have been shown to modulate aspects of cognitive performance. Whilst a number of studies have examined ginkgo's effects on electroencephalograph (EEG) recordings, to date, none have investigated the EEG effects of ginseng. In this double-blind, placebo-controlled, balanced crossover experiment, the effects of single doses of G. biloba (360 mg GK501), P. ginseng (200 mg G115), and an identical placebo, on auditory-evoked potentials, contingent negative variation (CNV), and resting power within the delta, theta, alpha, and beta wavebands, were assessed in 15 healthy volunteers. Each participant was assessed on three separate occasions 4 h after consuming that day's treatment. The order of presentation of the treatments was dictated by a Latin square with 7 days between testing sessions. The results showed that ginseng led to a significant shortening of the latency of the P300 component of the evoked potential. Both ginseng and ginkgo also led to significant reductions in frontal 'eyes closed' theta and beta activity, with additional reduction for ginseng in the alpha waveband. These findings demonstrate for the first time that P. ginseng can directly modulate cerebroelectrical activity, and that these effects are more pronounced than those following G. biloba. PMID- 12895689 TI - Effects on cognition and mood in postmenopausal women of 1-week treatment with Ginkgo biloba. AB - In a double-blind, placebo-controlled study, postmenopausal women (53-65 years old) were randomly assigned to 7-day treatment with Ginkgo (120 mg/day, n=15) or matched placebo (n=16). They were given a battery of cognitive tests and measurements of mood and menopausal symptoms at baseline (before treatment began) and at the end of 7 days. The group treated with Ginkgo was significantly better than the placebo group in a matching-to-sample test of nonverbal memory, but the groups did not differ in immediate or delayed paragraph recall or in delayed recall of pictures. In a test of frontal lobe function (rule shifting) and in the Paced Auditory Serial Addition Test (PASAT) (which measures sustained attention but also involves frontal lobe function), the group treated with Ginkgo performed significantly better than the placebo group. However, the groups did not differ in a test of planning. The treatments did not differ in their effects on the volunteers' ratings of menopausal symptoms, sleepiness, bodily symptoms or aggression. The benefits of Ginkgo on memory and frontal lobe function found in this study are modest but are unlikely to be secondary to major mood changes. PMID- 12895690 TI - Improved cognitive function in postmenopausal women after 12 weeks of consumption of a soya extract containing isoflavones. AB - We previously reported that a high soya diet improved memory and frontal lobe function in young volunteers, and since soya isoflavones are agonists at oestrogen receptors, they may improve these functions in postmenopausal women. Thirty-three postmenopausal women (50-65 years) not receiving conventional hormone replacement therapy (HRT) were randomly allocated in a double-blind parallel study to receive a soya supplement (60 mg total isoflavone equivalents/day) or placebo for 12 weeks. They received a battery of cognitive tests and completed analogue rating scales of mood and sleepiness, and a menopausal symptoms questionnaire before the start of treatment and then after 12 weeks of treatment. Those receiving the isoflavone supplement showed significantly greater improvements in recall of pictures and in a sustained attention task. The groups did not differ in their ability to learn rules, but the isoflavone supplement group showed significantly greater improvements in learning rule reversals. They also showed significantly greater improvement in a planning task. There was no effect of treatment on menopausal symptoms, self ratings of mood, bodily symptoms or sleepiness. Thus, significant cognitive improvements in postmenopausal women can be gained from 12 weeks of consumption of a supplement containing soya isoflavones that are independent of any changes in menopausal symptoms, mood or sleepiness. PMID- 12895691 TI - DNA encapsidation as a target for anti-herpesvirus drug therapy. AB - The current repertoire of approved anti-herpesviral drugs consists primarily of nucleoside analogues that inhibit viral replication by targeting the virus encoded DNA polymerase. This class of agents has been critical in controlling infections by herpes simplex, varicella zoster, and cytomegalovirus. However, because nucleoside analogues share a similar mechanism of action, treatment options are limited once resistance develops. This becomes an important medical issue with respect to the treatment of disease caused by resistant viral strains, particularly in immunocompromised individuals. Furthermore, several of the currently available therapies can result in mild to severe side effects making the discovery of less toxic drugs desirable. Efforts over the last decade have focused on the identification and development of improved therapies including less toxic compounds with novel mechanisms of action. Here we review the progress that has been made in targeting the DNA packaging and encapsidation process as a novel target for chemotherapy. Several recently identified compounds may warrant further development as a medically important group of herpesviral encapsidation inhibitors. PMID- 12895692 TI - Activation of latent HIV-1 expression by the potent anti-tumor promoter 12 deoxyphorbol 13-phenylacetate. AB - Agents that induce HIV-1 out of latency would be useful adjuvants for currently available anti-retroviral therapy. We report that nanomolar concentrations of 12 deoxyphorbol 13-phenylacetate (DPP), an anti-tumor-promoting phorbol ester originally isolated from a West African plant, induce the expression of HIV-1 in latently infected T cells and render them sensitive to killing by an immunotoxin targeted to the viral envelope glycoprotein. DPP also regulates an extensive series of genes under the control of protein kinase C, including several involved in T cell activation and cytoskeleton reorganization, and represses expression of the HIV-1 receptor CD4 and coreceptor CXCR4. DPP is 20-40-fold more potent than the related phorbol ester prostratin, probably due to its more lipophilic side chain structure. The combination of high potency and anti-tumor promoting activity make DPP an attractive candidate for the adjunctive therapy of persistent HIV-1 infection. PMID- 12895693 TI - Prevention of HIV-1 infection by phthalocyanines. AB - The ability of selected phthalocyanines and metallophthalocyanines to block HIV infection has been evaluated in an epithelial HeLa-CD4 cell line with an integrated LTR-beta-galactosidase gene. Sulfonated phthalocyanine itself (PcS), as well as its copper, nickel, and vanadyl chelates, were the most effective in blocking viral infection. These compounds were also very effective in blocking the fusion activity of the viral Env proteins. All of these compounds are expected to bind axial ligands weakly or not at all. In contrast, sulfonated phthalocyanines bearing metals expected to bind axial ligands more tightly (aluminum, cobalt, chromium, iron, silicon, and zinc) were less effective in blocking HIV infection and also less effective at inhibiting fusion. A number of active compounds were found to block binding of gp120 to CD4. Selected cationic and carboxy phthalocyanines, as well as porphyrazines, were also evaluated. Our results indicate that at least some of the compounds render the virus noninfectious, i.e. that they are virucidal. These compounds have potential as microbicides that might be used to provide protection against sexually transmitted HIV. PMID- 12895694 TI - Murine cytomegalovirus retinitis during retrovirus-induced immunodeficiency (MAIDS) in mice: interleukin-2 immunotherapy correlates with increased intraocular levels of perforin mRNA. AB - Mice with a retrovirus-induced immunosuppression (MAIDS) are susceptible to experimental murine cytomegalovirus (MCMV) retinitis, but can be rendered resistant to retinitis by systemic interleukin-2 (IL-2) immunotherapy. Experiments were performed to explore the mechanism by which IL-2 treatment during MAIDS might restore resistance to MCMV retinitis. Whereas 80% of untreated MAIDS mice were susceptible to MCMV retinitis, none (0%) of IL-2-treated MAIDS mice developed necrotizing retinitis. In comparison, 100% of both untreated and IL-2-treated perforin knockout mice (PKO mice) were susceptible to MCMV retinitis, and severity of retinitis and amounts of infectious intraocular MCMV in IL-2-treated PKO mice were equivalent to that in untreated PKO mice. A competitive quantitative RT-PCR assay was used to measure the levels of perforin mRNA within MCMV-infected eyes of immunologically normal mice, untreated MAIDS mice, and IL-2-treated MAIDS mice. Although the level of perforin mRNA within MCMV-infected eyes of untreated MAIDS mice susceptible to retinitis was significantly reduced when compared to the high level found within MCMV-infected eyes of normal mice resistant to retinitis, systemic treatment of MAIDS mice with IL-2 increased perforin mRNA within MCMV-infected eyes to levels found in normal mice. The ability of IL-2 treatment to increase intraocular levels of perforin mRNA diminished with the progression of MAIDS. Our findings support the hypothesis that systemic IL-2 immunotherapy during MAIDS provides protection against MCMV retinitis by upregulation of perforin-mediated cytotoxicity used by cytotoxic lymphocytes to kill virus-infected cells. PMID- 12895695 TI - The acyclic nucleoside phosphonate analogues, adefovir, tenofovir and PMEDAP, efficiently eliminate banana streak virus from banana (Musa spp.). AB - We report that the anti-retroviral and anti-hepadnavirus molecules, adefovir, tenofovir and 9-(2-phosphonomethoxyethyl)-2,6-diaminopurine (PMEDAP), efficiently eradicate the episomal form of Banana streak virus (BSV) from banana plants. Up to 90% of plants regenerated from BSV-infected highly proliferating meristems were virus free following a 6-month treatment period with 10 microg/ml (a non phytotoxic concentration) of either compounds. PMID- 12895696 TI - The potential molecular targets of marine sulfated polymannuroguluronate interfering with HIV-1 entry. Interaction between SPMG and HIV-1 rgp120 and CD4 molecule. AB - The potential targets of marine sulfated polymannuroguluronate (SPMG) involved in inhibition of HIV-1 entry were investigated by surface plasmon resonance and flow cytometry. Results indicated that binding of SPMG either to soluble oligomeric rgp120 or to complexed rgp120-sCD4 mainly resided in V3 loop region. In addition, SPMG was shown to be less accessible for sCD4 when sCD4 had pre-interacted with rgp120, though SPMG per se multivalently bound to sCD4 with relatively low affinity. While the pre-incubation of SPMG with rgp120 caused a partial blockade of rgp120 binding to sCD4, suggesting that SPMG either shared common binding sites on gp120 with sCD4 or masked the docking sites of gp120 for sCD4. Taken together, V3 domain was demonstrated to be the major site mediating interaction of SPMG with complexed rgp120-sCD4. It seems likely that SPMG binds to both rgp120 and sCD4, but has less accessibility for sCD4 when sCD4 has already bound to rgp120. Nevertheless, addition of SPMG either prior to or after the interaction of rgp120 with sCD4 may suppress rgp120 binding to sCD4. The exact pattern of this trimolecular complex formation at the cell membrane-anchored virus level requires further clarification. PMID- 12895697 TI - HIV-1 resistance to the gp41-dependent fusion inhibitor C-34. AB - The gp41 subunit of HIV-1 has been recently recognized as a target for antiviral therapy. C-34 is a peptide that mimics the heptad repeat 2 in the ectodomain of gp41. Here, we describe two HIV-1 strains selected after 5 and 17 passages in culture with increasing concentrations of C-34 (breakthrough concentration of 10 microg/ml). The HXB2-derived strain was more than 1000-fold resistant and contained a V38E mutation in the gp41 coding DNA sequence. The NL4-3-derived strain was more than 500-fold resistant and contained a L33S mutation in gp41. No cross-resistance to the RT inhibitor AZT, the HIV binding inhibitor dextran sulfate (DS), or the chemokine receptor antagonist ALX-40-4C was detected. These data indicate that HIV-1 can overcome C-34 inhibition through mutations at residues not involved in the formation of the hydrophobic cavity of gp41. PMID- 12896805 TI - Potent competitive inhibition of drug binding to the Saccharomyces cerevisiae ABC exporter Pdr5p by the hydrophobic estradiol-derivative RU49953. AB - The hydrophobic estradiol-derivative RU49953 inhibits the energy-dependent interaction of yeast multidrug-transporter Pdr5p with its fluorescent drug substrate rhodamine 6G. The potent inhibition is competitive towards drug binding (Ki=23+/-6 nM), whereas nucleoside-triphosphate hydrolysis is two-orders-of magnitude less sensitive. RU49953 constitutes the most efficient inhibitor of drug binding to a yeast multidrug ABC exporter reported so far. PMID- 12896806 TI - Disc formation in cholesterol-free liposomes during phase transition. AB - Cryogenic transmission electron microscopy (cryo-TEM) images of lysolipid containing thermosensitive liposomes (LTSL) revealed that open liposomes and bilayer discs appeared when liposomes were cycled through the gel (Lbeta') to liquid-crystalline (Lalpha) phase transition. The amount of bilayer discs generated was dependent on the combined presence of PEG-lipid and lysolipid in the membrane. We hypothesize that micelle-forming membrane components stabilize the rim of bilayer openings and membrane discs that form when liposomes are cycled through TC. PMID- 12896807 TI - The role of ubiquitin in down-regulation and intracellular sorting of membrane proteins: insights from yeast. AB - Ubiquitination is a versatile tool used by all eukaryotic organisms for controlling the stability, function, and intracellular localization of a wide variety of proteins. Two of the best characterized functions of protein ubiquitination are to mark proteins for degradation by cytosolic proteasome and to promote the internalization of certain plasma membrane proteins via the endocytotic pathway, followed by their degradation in the vacuole. Recent studies of membrane proteins both in yeast and mammalian cells suggest that the role of ubiquitin may extend beyond its function as an internalization signal in that it also may be required for modification of some component(s) of the endocytotic machinery, and for cargo protein sorting at the late endosome and the Golgi apparatus level. In this review, I will attempt to bring together what is currently known about the role of ubiquitination in controlling protein trafficking between the yeast plasma membrane, the trans-Golgi network, and the vacuole/lysosome. PMID- 12896808 TI - Biological activity and characteristics of triamcinolone-acetonide formulated with the self-regulating drug carriers, Transfersomes. AB - Novel formulations of the halogenated corticosteroid, triamcinolone-acetonide, based on ultradeformable mixed lipid vesicles, Transfersomes, are described. Their performance was tested in vivo using radioactive label measurements, to study the drug biodistribution, and murine ear edema, to determine the drug bioactivity. Sparse use of drug-loaded Transfersomes on the skin ensures an almost exclusive delivery of triamcinolone-acetonide into the organ, thus arguably increasing the treatment safety. Delivery of triamcinolone-acetonide in the skin with ultradeformable vesicles prolongs the anti-inflammatory drug action several times compared to drug usage in a conventional creme or an ointment, the robustness of biological response for the former being at least identical to the latter. The required dose of Transfersome-based triamcinolone-acetonide is also greatly reduced. The drug dose of 0.2 microg cm(-2) suppresses 75% of arachidonic acid-induced murine ear edema for at least 48 h. In contrast, a conventional formulation of triamcinolone-acetonide requires a 10-fold higher drug dosage to achieve a similar effect. In either case, increasing the applied corticosteroid amount delays the onset of anti-edema action. PMID- 12896809 TI - Shifts in chain-melting transition temperature of liposomal membranes by polymer grafted lipids. AB - The chain-melting transition temperature of dipalmitoyl phosphatidylcholine (DPPC) bilayer membranes containing poly(ethylene glycol)-grafted dipalmitoyl phosphatidylethanolamine (PEG-DPPE) was determined by optical turbidity measurements. The dependence on content, Xp, of PEG-DPPE lipid was studied for different polar headgroup sizes, np, of the polymer lipid, throughout the lamellar phase of the mixtures with DPPC. Mean-field theory for the polymer brush regime predicts that the downward shift in transition temperature should vary with polymer size and content as npXp(5/3) (approximately npXp(11/6) for scaling theory). Any shift induced by the charge on PEG-lipids is independent of polymer size. These predictions are reasonably borne out for the longer polymer lipids (PEG molecular masses 750, 2000 and 5000 Da). Transition temperature shifts in the lamellar phase, before the onset of micellisation, are in the region of -1 to -2 degrees C (+/-0.1-0.2 degrees C) in reasonable accord with theoretical estimates of the lateral pressure exerted by the polymer brush. Shifts of this size are significant to the design of liposomes for controlled release of contents by mild hyperthermia. PMID- 12896810 TI - The influence of alkyl pyridinium sponge toxins on membrane properties, cytotoxicity, transfection and protein expression in mammalian cells. AB - The ability of two alkyl pyridinium sponge toxin preparations (poly-APS and halitoxin) to form transient pores/lesions in cell membranes and allow transfection of plasmid cDNA have been investigated using HEK 293 cells. Poly-APS and halitoxin preparations caused a collapse in membrane potential, reductions in input resistance and increased Ca2+ permeability. At least partial recovery was observed after poly-APS application but recovery was more rarely seen with halitoxin. The transfection with plasmid cDNAs for an enhanced green fluorescent protein (EGFP) and human tumour necrosis factor receptor 2 (TNFR2) was assessed for both toxin preparations and compared with lipofectamine. Stable transfection was achieved with poly-APS although it was less efficient than lipofectamine. These results show that viable cells transfected with alien cDNA can be obtained using novel transient pore-forming alkyl pyridinium sponge toxins and a simple pre-incubation protocol. This provides the first proof of principle that pore forming alkyl pyridinium compounds can be used to deliver cDNA to the intracellular environment without permanently compromising the plasma membrane. PMID- 12896811 TI - The use of fluorescence resonance energy transfer to monitor dynamic changes of lipid-DNA interactions during lipoplex formation. AB - Fluorescence resonance energy transfer (FRET) was used to monitor interactions between Cy3-labeled plasmid DNA and NBD-labeled cationic liposomes. FRET data show that binding of cationic liposomes to DNA occurs immediately upon mixing (within 1 min), but FRET efficiencies do not stabilize for 1-5 h. The time allowed for complex formation has effects on in vitro luciferase transfection efficiencies of DOPE-based lipoplexes; i.e., lipoplexes prepared with a 1-h incubation have much higher transfection efficiencies than samples with 1-min or 5-h incubations. The molar charge ratio of DOTAP to negatively charged phosphates in the DNA (DOTAP+/DNA-) also affected the interaction between liposomes and plasmid DNA, and interactions stabilized more rapidly at higher charge ratios. Lipoplexes formulated with DOPE were more resistant to high ionic strength than complexes formulated with cholesterol. Taken together, our data demonstrate that lipid-DNA interactions and in vitro transfection efficiencies are strongly affected by the time allowed for complex formation. This effect is especially evident in DOPE-based lipoplexes, and suggests that the time allowed for lipoplex formation is a parameter that should be carefully controlled in future studies. PMID- 12896812 TI - Role of pulse shape in cell membrane electropermeabilization. AB - The role of the amplitude, number, and duration of unipolar rectangular electric pulses in cell membrane electropermeabilization in vitro has been the subject of several studies. With respect to unipolar rectangular pulses, an improved efficiency has been reported for several modifications of the pulse shape: separate bipolar pulses, continuous bipolar waveforms, and sine-modulated pulses. In this paper, we present the results of a systematic study of the role of pulse shape in permeabilization, cell death, and molecular uptake. We have first compared the efficiency of 1-ms unipolar pulses with rise- and falltimes ranging from 2 to 100 micros, observing no statistically significant difference. We then compared the efficiency of triangular, sine, and rectangular bipolar pulses, and finally the efficiency of sine-modulated unipolar pulses with different percentages of modulation. We show that the results of these experiments can be explained on the basis of the time during which the pulse amplitude exceeds a certain critical value. PMID- 12896813 TI - Molecular modeling of the bacterial outer membrane receptor energizer, ExbBD/TonB, based on homology with the flagellar motor, MotAB. AB - The MotA/MotB proteins serve as the motor that drives bacterial flagellar rotation in response to the proton motive force (pmf). They have been shown to comprise a transmembrane proton pathway. The ExbB/ExbD/TonB protein complex serves to energize transport of iron siderophores and vitamin B12 across the outer membrane of the Gram-negative bacterial cell using the pmf. These two protein complexes have the same topology and are homologous. Based on molecular data for the MotA/MotB proteins, we propose simple three-dimensional channel structures for both MotA/MotB and ExbB/ExbD/TonB using modeling methods. Features of the derived channels are discussed, and two possible proton transfer pathways for the ExbBD/TonB system are proposed. These analyses provide a guide for molecular studies aimed at elucidating the mechanism by which chemiosmotic energy can be transferred either between two adjacent membranes to energize outer membrane transport or to the bacterial flagellum to generate torque. PMID- 12896814 TI - Zinc uptake across the apical membrane of freshwater rainbow trout intestine is mediated by high affinity, low affinity, and histidine-facilitated pathways. AB - Zinc is both a vital nutrient and an important toxicant to aquatic biota. In order to understand the interplay between nutrition and toxicity, it will be important to determine the mechanisms and the factors that regulate zinc uptake. The mechanism of apical intestinal Zn(II) uptake in freshwater rainbow trout and its potential modification by the complexing amino acid histidine was investigated using brush-border membrane vesicles (BBMVs). Following characterisation of the BBMV preparation, zinc uptake in the absence of histidine was both time- and concentration-dependent and consisted of two components. A saturable phase of uptake was described by an affinity constant of 57+/-17 microM and a transport capacity of 1867+/-296 nmol mg membrane protein(-1) min(-1). At higher zinc levels (>500 microM) a linear, diffusive component of uptake was evident. Zinc transport was also temperature-dependent, with Q10 values suggesting zinc uptake was a carrier-mediated process. Zinc uptake by vesicles in the presence of histidine was correlated to a mono-histidine species (Zn(His)+) at all Zn(II) concentrations examined. PMID- 12896815 TI - Preeclampsia and human reproduction. An essay of a long term reflection. AB - Hypertensive disorders of pregnancy (HDP: pregnancy-induced hypertension, preeclampsia, eclampsia) affect approximately 10% of human births. Women are at increased risk for HDP during their first conception; and/or when the conception is with a new partner (new paternity); when conception occurs very shortly after the beginning of their sexual relationship. A primary cause of preeclampsia is the defect of the normal human-specific deep endovascular invasion of trophoblast, which is a consequence of the nutritional demands of growth of the human fetal brain. The occurrence of preeclampsia represents a reproductive disadvantage unique to humans compared with other mammals. As such, it may have played a significant role in shaping human reproduction and, therefore, human sexuality. This deep implantation/preeclampsia phenomenon may explain many anthropological mysteries of human sexuality that do not exist in other mammalian species (and primates). These include: very low fertility rate, concealed ovulation, all year long 'apparent-waste-of-efficiency' sexuality, absence of sperm competition in human females at the time of conception, and the unexplained testicle size in human males compared with relevant primates. Further, this deep trophoblastic implantation (and its failure in preeclampsia) in humans might be a decisive condition of hominization between great apes and all the other Homo genuses. This frontier might even have occurred inside these Homo lineages: because of their relatively small brains, the first species of Homo might not have presented the deep trophoblastic invasion described in Homo sapiens. PMID- 12896816 TI - Epidemiology of preeclampsia in Mauritius island. AB - In Mauritius, the maternal mortality rate (MMR) was 21 per 100000 live births for the year 2001 and this is comparable with figures from developed countries. There has been poor documentation in the area of hypertensive disorders of pregnancy until recently in the island. Consequently, we analysed data from the Ministry of Health. Hypertensive disorders of pregnancy occurred in 6.7% gravidas in 2001. Out of four maternal deaths recorded in 2001, two occurred due to severe eclampsia. In the second part of our study, we scrutinised also the obstetric notes and interviewed 862 hospitalised women from four main hospitals of the island between 1995 and 1997. The majority of women were affected by non proteinuric hypertension (70%), followed by preeclampsia (24%) and chronic hypertension (6%). Preeclamptic primi and multigravidas developed the condition earlier and also gave birth to growth-retarded babies. Highly significant results supported the relationship between growth retardation and preeclampsia in different parities, irrespective of the length of exposure of the baby in utero. Preeclampsia affects both Indo-Mauritians and Creoles to the same extent, but the latter are much more prone to gestational hypertension. In addition, hypertensive disorders of pregnancy are more a problem of multigravid women in Creoles while they are more classically a condition of primigravidae in Indians. PMID- 12896817 TI - Human reproduction: a comparative background for medical hypotheses. AB - Successful reconstruction of any aspect of human evolution requires broad-based comparisons with other primates and other eutherian mammals. If quantitative data are involved, the scaling influence of body size must be taken into account, and bivariate allometric analysis is a very useful tool. Broad-based comparisons lead to recognition of general principles that can feed into hypotheses concerning human reproductive medicine. Here, this approach is applied to basic life-history parameters (age at sexual maturity, gestation period, litter size, lactation period, interbirth interval, maximum lifespan), testicular dimensions and male mating strategies, female ovarian processes and mating cycles, placentation and embryonic/fetal development, gestation periods and neonatal condition, and brain development in relation to reproduction. Comparisons reveal that some features claimed to be unique to human beings in fact occur more widely in primates, such as 'loss of oestrus', which probably evolved in the common ancestor of higher primates (monkeys, apes and humans). Although humans show an extreme condition, in most higher primates mating is not strictly confined to the periovulatory period. Because this condition introduces the danger of fertilization with ageing gametes, it must have been favoured by strong selection and accompanied by the evolution of compensatory mechanisms. Overall, it seems likely that the high incidence of pre-eclampsia in human pregnancies is attributable to a partial failure of compensatory mechanisms associated with highly invasive placentation. PMID- 12896818 TI - Increased cranial capacity in hominid evolution and preeclampsia. AB - One of the major trends in primate evolution generally and hominid evolution in particular, is cranio-facial contraction accompanied by an increase in cranial capacity. Landmark-based morphometric methods are applied to adult skulls of great apes (Gorilla, Pan), australopithecines (Australopithecus and Paranthropus), and humans (Homo eragster, erectus, neanderthalensis, and sapiens). Morphological changes quantified by vector fields (Procrustes methods) indicate that these skull plans are characterized by distinctive degrees of cranio-facial contraction. These suggest the existence of three discrete skull organization plans: "great ape", "australopithecine" and "Homo". This paper focuses on the "Homo" skull bauplan and discusses the possible relationships between greatly increased cranial capacity and preeclampsia. The earliest species of the human lineage exhibit less cranio-facial contraction and smaller cranial capacity than Homo neanderthalensis and modern Homo sapiens. Neandertalization introduces a posterior elongation of the skull and leads to a large increase in cranial capacity in the last Neandertals, with values as large as in present-day H. sapiens. Consequently, a new biological hypothesis is proposed to account for the unexplained disappearance of H. neanderthalensis some 30000 years ago related to the possible appearance of preeclampsia as a factor affecting the survival of the species. PMID- 12896819 TI - Trophoblast deportation and the maternal inflammatory response in pre-eclampsia. AB - We have proposed that the maternal syndrome of pre-eclampsia is caused by a systemic inflammatory response involving both leucocytes and endothelium. This inflammatory response is present also in normal pregnancy, but in a milder form. The inflammatory stimulus is most likely to come from the placenta. Syncytiotrophoblast apoptotic debris, which is shed into the maternal circulation in normal pregnancy and in increased amounts in pre-eclampsia, may be the stimulus for this response. It may also contribute to the suppression of Th1 responses seen in pregnancy. PMID- 12896820 TI - Th1/Th2 balance in preeclampsia. AB - The syndrome of preeclampsia has previously been ascribed to generalized maternal endothelial dysfunction, poor placentation and excessive maternal inflammatory response. Recent reports suggest that preeclampsia is associated with a Th1 predominant profile and may be considered as a failure of the tolerance system allowing the second physiological trophoblastic invasion. In this review, we discuss that Th1 predominant immunity is closely related to inflammation, endothelial dysfunction and poor placentation. PMID- 12896821 TI - Update on pathways regulating the activation of uterine Natural Killer cells, their interactions with decidual spiral arteries and homing of their precursors to the uterus. AB - Virgin adult C57Bl/6J mouse uterus contains a population of small, non-granulated Natural Killer (NK) cells with balanced expression of NK cell activating and inhibiting LY49 receptors. Coincident with blastocyst implantation and decidualization, uterine (u)NK cells become activated. The surface glycoslyation of uNK changes, the cells proliferate and they induce production of interferon (IFN)gamma, perforin, serine esterases and other molecules, including angiogenic factors. Mouse strains genetically ablated in uNK cells fail to undergo modification of spiral artery segments that branch from the uterine artery and feed into the placenta and these mice do not sustain a robust decidualization response. IFN-gamma is thought, from bone marrow transplantation and therapeutic studies, to be the key uNK-cell derived mediator regulating gene expression in vascular and decidual tissues. Here, we review recent studies showing that IL-15 is the critical cytokine controlling uNK cell differentiation and that uNK cells are activated by either IL-12 or IL-18 and by other factors when both IL-12 and IL-18 are genetically absent from implantation sites. We address possible roles of the IFN-gamma regulated gene alpha2-macroglobulin (alpha2-M) in regulation of the position of fetal trophoblast within the walls of the spiral arteries, and we discuss approaches that have been successful in evaluating mechanisms involved in homing of mouse uNK cell precursors to the uterus. These approaches maybe applicable to studies in women. Our studies show that complex immuno physiological events contribute to spiral artery modification by mid-gestation in mice. PMID- 12896822 TI - Blood myeloid and lymphoid dendritic cells are stable during the menstrual cycle but deficient during mid-gestation. AB - The aim of this study was to estimate the populations of peripheral blood myeloid and lymphoid dendritic cells (CD1c(+), BDCA-2(+), BDCA-4(+)) and the CD1c(+):BDCA 2(+) ratio in phases of the ovarian cycle and in normal pregnant patients. 18 non pregnant women and 17 normal pregnant women were included. Dendritic cells were isolated from peripheral blood, stained with monoclonal antibodies (mAbs) against blood dendritic cell antigens (anti-BDCA-1, BDCA-2, BDCA-4) and estimated using flow cytometry. CD1c(+), BDCA-2(+) and BDCA-4(+) dendritic cells were present in the follicular and luteal phases of the ovarian cycle and in all trimesters of normal pregnancy. The percentages of CD1c(+) dendritic cells did not differ between the follicular and luteal phases of the ovarian cycle. The percentage of BDCA-2(+) dendritic cells was lower in the luteal phase of the ovarian cycle compared with the follicular phase, but the differences were not statistically significant. The CD1c(+):BDCA-2(+) cell ratio was significantly lower in the luteal phase compared with the follicular phase of the ovarian cycle. The numbers of dendritic cells were significantly lower in the second trimester when compared with the first and third trimesters of normal pregnancy. Furthermore, in the second trimester, the CD1c(+):BDCA-2(+) ratio was higher than in the other trimesters of normal pregnancy. All populations of dendritic cells and the CD1c(+):BDCA-2(+) ratio did not differ in the first and third trimesters of physiological pregnancy. Our results suggest that myeloid and lymphoid dendritic cells are not affected by steroid hormones during the menstrual cycle. The deficiency of peripheral blood dendritic cells observed during the second trimester of normal pregnancy can be associated with their migration to the uterus during the second physiological invasion by cytotrophoblast. PMID- 12896823 TI - Implantation: can immunological parameters of implantation failure be of interest for pre-eclampsia? AB - We restate briefly why we consider that the Th1/Th2 paradigm, as useful as it has been, is now no longer adequate and is obsolete. We take as an example the role of IL-18, abortifacient at high doses but cardinal for the control of natural killer (NK) cell effects on spiral artery remodelling in mice, and likely also in humans. We then describe briefly our recent studies on cytokine defects and implantation failure in humans, a key feature being the link between uterine cytokine dysregulation and abnormal uterine vascular scores. We draw lessons for preeclampsia, and describe features of a model for its immune aetiology. PMID- 12896824 TI - HLA-G, pre-eclampsia, immunity and vascular events. AB - Pre-eclampsia, one of the main complications in pregnancy, is characterised by shallow cytotrophoblast invasion of decidua as well as by vascular endothelial cell dysfunction, leading to a poor perfusion of placenta. A striking feature of pre-eclamptic pregnancies is that expression of HLA-G protein is reduced in term placentas compared with normal pregnancy. How such HLA-G deficient expression may be related to the pre-eclamptic pathology is unknown. Here, we review the major structural characteristics of HLA-G and some of its functions that have been recently characterised. Soluble HLA-G1 isoform down-regulates both CD8(+) and CD4(+) T cell reactivity. HLA-G also modulates innate immunity by binding to several NK and/or decidual receptors, inducing particular cytokine secretion. HLA G was shown to be less susceptible to human cytomegalovirus-derived US protein down-modulation. Finally, soluble HLA-G1 down-regulates endothelial cell proliferation and migration. In view of these different HLA-G properties, we will briefly discuss how defective HLA-G function may contribute to the low trophoblast invasion and vascular abnormalities observed in pre-eclamptic placentas. PMID- 12896825 TI - HLA-DR in couples associated with preeclampsia: background and updating by DNA sequencing. AB - The placenta acts as an immunological barrier between the mother and the fetal "graft", allowing two antigenically different organisms to tolerate one another. In placentae from preeclamptic women, we have demonstrated, by an ultrastructural assessment and an immunohistochemical study, a placental barrier breakage leading to the mixing of maternal and fetal antigenically different blood. This condition could be responsible for the triggering of a maternal rejection reaction that we presume to be at the basis of the preeclamptic syndrome. Thus, we have investigated the Human Leukocyte class II DR antigens (HLA-DR), whose role in self and non-self recognition is well known, in women with preeclampsia, their partners and in control couples using the serological Terasaki tecnique. The results showed a statistically significant increase of HLA-DR homozygosity and a reduced antigenic variety in the preeclamptic women and their partners with respect to controls. In this update, we have examined the 2nd exon of the human gene, HLA-DRB1, on the short arm of the chromosome 6 using DNA sequence-based typing (S-BT) PCR in 56 preeclamptic couples and 64 control couples. The results have confirmed the significant excess of HLA-DR homozygosity in couples associated with preeclampsia versus controls. From our results, it emerges that HLA-DR homozygosity and the reduced antigenic variety seem to be associated to a major risk for preeclampsia, which further appears to be a "couple's disease". PMID- 12896826 TI - The birth interval hypothesis-does it really indicate the end of the primipaternity hypothesis. AB - Recent Norwegian data suggest the presence of a causal relationship between prolonged birth intervals and the risk for preeclampsia in subsequent pregnancies. It has been proposed that the birth interval data explain the known association between a change in paternity and the risk for preeclampsia. In this review, the authors explore alternative explanations for the Norwegian findings, and as such argue that there is currently no reason to reject the primipaternity hypothesis. PMID- 12896827 TI - Seminal 'priming' for protection from pre-eclampsia-a unifying hypothesis. AB - Conventional belief holds that an immune response to ejaculate antigens should interfere with fertilisation and establishment of pregnancy. However, emerging evidence now supports the opposing view-that insemination acts to activate maternal immune mechanisms exerting a positive effect on reproductive events. In a response well documented in rodents, semen triggers an influx of antigen presenting cells into the female reproductive tract which process and present paternal ejaculate antigens to elicit activation of lymphocytes in the adaptive immune compartment. Transforming growth factor beta (TGFbeta), a cytokine present in abundance in seminal plasma, initiates this inflammatory response by stimulating the synthesis of pro-inflammatory cytokines and chemokines in uterine tissues. Lymphocyte activation is evident in lymph nodes draining the uterus and leads to hypo-responsiveness in T-cells reactive with paternal alloantigens. TGFbeta has potent immune-deviating effects and is likely to be the key agent in skewing the immune response against a Type-1 bias. Prior exposure to semen in the context of TGFbeta can be shown to be associated with enhanced fetal-placental development late in gestation. In this paper, we review the experimental basis for these claims and propose the hypothesis that, in women, the partner-specific protective effect of insemination in pre-eclampsia might be explained by induction of immunological hypo-responsiveness conferring tolerance to histocompatibility antigens present in the ejaculate and shared by the conceptus. PMID- 12896828 TI - Possible involvement of inositol phosphoglycan-P in human parturition. AB - Preterm labour is a major cause of neonatal morbidity and mortality but the pathophysiology that underlies preterm labour is unknown. Inositolphosphoglycans (IPGs) comprise a ubiquitous family of putative carbohydrate second messengers and they have been linked to the pathogenesis of various conditions, including diabetes and pre-eclampsia. Studying IPG-P levels in normal and pre-eclamptic pregnancies, we noticed a constant rise of urinary IPG-P levels in all women at the time of delivery. A prospective pilot study of urinary IPG-P levels in 23 non labouring and labouring women with uncomplicated pregnancies has, therefore, been performed. Levels of urinary IPG-P were significantly higher in labour than in the non-labouring group (P<0.0001). These higher levels have been found in both spontaneous and induced labour. The clinical significance of this observation with particular reference to the onset of labour itself is discussed. PMID- 12896829 TI - Placental GPI-PLD is of maternal origin and its GPI substrate is absent from placentae of pregnancies associated with pre-eclampsia. AB - Pre-eclampsia (PE) is a disorder affecting 5-10% of all pregnancies and is characterised by abnormal trophoblast invasion, maternal endothelial cell dysfunction and a systemic maternal response. A unifying factor responsible for eliciting these effects remains unknown. However, levels of the autocrine insulin mediators, inositolphosphoglycans (IPG), are elevated 3-fold in pre-eclamptic placentae compared with controls and are also elevated 3-fold in maternal urine of pre-eclamptic women, suggesting an abnormal paracrine role of the mediator in the systemic maternal response. At the placental level, IPGs are metabolic second messengers capable of eliciting some of the characteristic features of PE, such as the 10-fold increase in glycogen synthesis and 16-fold increase in the activity of the IPG-dependent enzyme glycogen synthase. IPGs are derived from their lipidic precursors, the glycosylphosphatidylinositols (GPI), in membrane associated caveolae by the action of a GPI-specific phospholipase D whose activity is regulated by its membrane microenvironment. We show that the lipidic GPI precursor was detected in total placental membrane and microvillous membrane from normal placentae. The presence of GPI could not be detected in PE placentae, suggesting that the GPI/IPG signalling system is dysregulated in this disorder. Equivalent amounts of a proteolytically-cleaved 50 kDa GPI-PLD protein is detected in both normal and PE placentae. However, GPI-PLD mRNA is absent, suggesting a mechanism of uptake from maternal serum. Since GPI-PLD, whose presence is required for hydrolysis of GPI and release of free IPG, is detectable with equal activity in both normal and PE placentae, we postulate that dysregulation of the tubular caveolar structure of the microvilli in pre eclamptic placentae provides an environment which promotes the unregulated hydrolysis of GPI in this disorder. PMID- 12896830 TI - American Society for Mass Spectrometry directory of members and corporate members. PMID- 12896831 TI - Cu@Au alloy nanoparticle as oligonucleotides labels for electrochemical stripping detection of DNA hybridization. AB - Synthesis of the novel Cu@Au alloy nanoparticle and its application in an electrochemical DNA hybridization detection assay is described in this article. We report a low-temperature method for generating core-shell particles consisting of a core of Cu and a thin layer of Au shell that can be readily functionalized with oligonucleotides. Core-shell Cu@Au particles were successfully labeled to a 5'-alkanethiol capped oligonucleotides probe that is related to the colitoxin gene. The DNA genetic sensing assay relies on the electrostatic adsorption of target oligonucleotides onto conducting polypyrrole (PPy) surface at the glassy carbon electrode (GCE), and its hybridization to the alloy particle oligonucleotides DNA probe. Hybridization events between probe and target were monitored by the release of the copper metal atoms anchored on the hybrids by oxidative metal dissolution and the indirectly determination of the solubilized Cu2+ ions by sensitive anodic stripping voltammetry (ASV). The detection limit is 5.0 pmol l(-1) of target oligonucleotides. The Cu@Au core-shell nanoparticles combining the surface modification properties of Au with the good electrochemical activity of Cu core shows their perspective application in the electrochemical DNA hybridization analysis assay. PMID- 12896832 TI - A highly sensitive flow-through amperometric immunosensor based on the Peroxidase chip and enzyme-channeling principle. AB - A concept based on the Peroxidase-chip (P-chip), antibody co-immobilization, competitive and enzyme-channeling principle was exploited to develop an integrated flow-through amperometric biosensor for detection of environmental pollutants such as s-triazine herbicides. In this concept, recombinant peroxidase is immobilized on the gold electrode (P-chip) in such a way that direct electron transfer is achieved. The recognition and quantitation the target analyte is realized through the competition between the simazine-glucose oxidase (GOD) conjugate and free simazine for the binding sites of the monoclonal antibody co immobilized with peroxidase on the gold electrode. The arrangement allows to generate a specific signal in the presence of glucose through the channeling of H2O2 produced by GOD conjugate bound to the antibody. The immunosensor exhibited 50% signal decrease (IC50 value) at approximately 0.02 microg l(-1). A concentration of 0.1 ng l(-1) gave a signal clearly distinguishable from the blank whereas the ELISA using the same antibody had a typical detection limit of about 1 microg l(-1), which is four orders of magnitude higher compared to the presented biosensor system. The results demonstrated that gene engineering biomolecules, in this case recombinant peroxidase, might be attractive reagents for the development of electrochemical immunosensors. PMID- 12896833 TI - New principle of direct real-time monitoring of the interaction of cholinesterase and its inhibitors by piezolectric biosensor. AB - This paper describes a new method for the sensitive detection of cholinesterase inhibitors based on real-time monitoring using a piezoelectric biosensor. The cholinesterase inhibitor paraoxon was immobilized on the sensing surface via a chelate complex as the recognition element. At first, the conjugate of N mercaptoundecanoic acid (MUA) with Nalpha,Nalpha-bis (carboxymethyl)-L-lysine (NTA-Lys) was chemisorbed to form a self-assembled monolayer on the surface of the gold electrode of the piezosensor. In the next step, paraoxon-spacer hexahistidine conjugate was linked to the MUA-Lys-NTA layer via the chelate complex with Ni2+. The paraoxon-modified surface thus obtained was applied for the binding of human butyrylcholinesterase (BChE). Regeneration of the sensing surface was achieved by splitting the chelate complex with EDTA and depositing a fresh layer of Ni2+ followed by addition of the paraoxon-spacer-hexahistidine. In the presence of free inhibitors like diisopropylfluorophosphate (DFP), binding of BChE to the surface-bound paraoxon was decreased. In this way, a competitive affinity assay for organophosphorus compounds was developed. The limit of detection for DFP as a model compound was 10 nmol/l (ca. 2 microg/l). This new concept seems suitable for constructing biosensors for the group-specific detection of cholinesterase-inhibiting substances like insecticides in the field. PMID- 12896834 TI - A portable microelectrode array recording system incorporating cultured neuronal networks for neurotoxin detection. AB - Cultured neuronal networks, which have the capacity to respond to a wide range of neuroactive compounds, have been suggested to be useful for both screening known analytes and unknown compounds for acute neuropharmacologic effects. Extracellular recording from cultured neuronal networks provides a means for extracting physiologically relevant activity, i.e. action potential firing, in a noninvasive manner conducive for long-term measurements. Previous work from our laboratory described prototype portable systems capable of high signal-to-noise extracellular recordings from cardiac myocytes. The present work describes a portable system tailored to monitoring neuronal extracellular potentials that readily incorporates standardized microelectrode arrays developed by and in use at the University of North Texas. This system utilizes low noise amplifier and filter boards, a two-stage thermal control system with integrated fluidics and a graphical user interface for data acquisition and control implemented on a personal computer. Wherever possible, off-the-shelf components have been utilized for system design and fabrication. During use with cultured neuronal networks, the system typically exhibits input referred noise levels of only 4-6 microVRMS, such that extracellular potentials exceeding 40 microV can be readily resolved. A flow rate of up to 1 ml/min was achieved while the cell recording chamber temperature was maintained within a range of 36-37 degrees C. To demonstrate the capability of this system to resolve small extracellular potentials, pharmacological experiments with cultured neuronal networks have been performed using ion channel blockers, tetrodotoxin and tityustoxin. The implications of the experiments for neurotoxin detection are discussed. PMID- 12896835 TI - Real-time evaluation of macromolecular surface modified quartz crystal resonant sensors under cryogenic stress for biological applications. AB - This study presents a novel auto-gain-control based quartz acoustic sensor technology capable of constant quartz crystal operation when cycled between ambient (22 degrees C) and cryogenic temperatures (-196 degrees C), afforded by direct exposure of crystals to bulk liquid nitrogen. The real-time frequency response profiles due to freeze-thaw cycling on crystals of differing surface finish and two model macromolecular surface coatings were studied in order to determine surface events such as water uptake. The quartz crystal surface finishes used were optically polished or lapped to one of two surface finishes. These were used as control native gold electrodes, and these surfaces were further coated with bovine serum albumin or the tri-block copolymer, poloxamer 188 as model macromolecular surface architectures. Crystals were snap frozen in liquid nitrogen and allowed to return to ambient temperature under controlled conditions. The processes of ice formation, thawing and evaporation were followed in real-time and comparisons were made between the test samples in order to assess the capability of this technique for sensing changes in surface characteristics such as the entrapment of water. PMID- 12896836 TI - A new approach for immobilization of oligonucleotides onto piezoelectric quartz crystal for preparation of a nucleic acid sensor for following hybridization. AB - The aim of this study is to develop a nucleic acid sensor based on piezoelectric crystal microbalance system (QCM) for following hybridization. Piezoelectric quartz crystal surfaces were first treated in a glow-discharge apparatus with ethylene diamine (EDA) plasma at 15 W (discharge power), 2.5 min (incubation time) and 35 ml/min (monomer flow rate) to create amino groups on the crystal surfaces. The thickness of the EDA-plasma film formed was about 43+/-24 A. Then, the amino groups on the crystal surfaces were converted to aldehyde groups by reacting the amino groups with glutaraldehyde (GA) at different conditions. A GA concentration of 2.5% and an incubation time of 2 h were selected as optimal values at this step, corresponding to a GA surface concentration of about 270 ng/cm2. A double strand Oligonucleotides, having one extra base on 5'-end of one of the complementary strands, were immobilized through the amino groups of this base onto the GA-modified crystals. Optimal immobilization conditions were as follows: oligonucleotide concentration: 1 microg/ml; time: 3 h; pH: 9.2 carbonate buffer; ionic strength: 0.1; and temperature: 20 degrees C. The QCM sensor carrying the covalently bound strand was used in the hybridization experiments, which showed that equilibrium is achieved in about 5 min, and the frequency shift measured is related to the concentration of the target strand to be measured within the medium. PMID- 12896837 TI - An artificial taste sensor based on conducting polymers. AB - Pure and composite nanostructured films of conducting polymers were used as individual sensing units constituting an electronic tongue. The use of extremely thin films for signal transduction via impedance spectroscopy measurements in the frequency range 10-1 MHz allows the detection of trace amounts of tastants and inorganic contaminants in liquid systems. In addition, the sensor could detect the suppression of sourness by sweetness displaying similarities with the biological system. Brands of several commercial beverages could be easily distinguished without complex analysis, including the discrimination of waters, tastants and wines. PMID- 12896838 TI - A fiber optic biosensor (FOBS) to monitor mutans streptococci in human saliva. AB - A fiber optic biosensor (FOBS) to monitor mutans streptococci activity in human saliva is developed. The biosensor utilizes e fiber optic evanescent wave spectroscopy to monitor a bacterial mediated biochemical reaction. To achieve this, a short length of the cladding is removed; the fiber core surface is treated and coated with a thin film of porous glass medium using sol-gel technique. The mutans streptococci mediated reaction with sucrose is monitored using a photosensitive indicator, which is immobilized within the porous glass coating. Spectroscopic analysis shows that the transmitted intensity at 597 nm increases conspicuously when monitored for 120 min. Two distinct phases are observed, one from 0 to 60 min and the other from 60 to 120 min. A negative correlation coefficient between the rate of increase in absorption peak intensity recorded by the FOBS and the decrease in pH measured using the pH meter, was calculated to be rho=-0.994. This investigation highlights the potential benefits of this sensor to monitor mutans streptococci activity in saliva. PMID- 12896839 TI - Monitoring the cellular activity of a cultured single cell by scanning electrochemical microscopy (SECM). A comparison with fluorescence viability monitoring. AB - The respiratory activities of cultured HeLa cells were monitored at a single cell level using scanning electrochemical microscopy (SECM) that produces images of the localized distribution of oxygen around the cell. The change in the cellular activity was traced after exposures to KCN, ethyl alcohol and the antibiotic drug, Antimycin A. The results were compared with those from the conventional fluorescence monitoring using Calcein-AM that is sensitive to deformation of the cell membrane. The SECM-based measurement follows the decrease in the cellular activity upon exposure to KCN and Antimycin A more rapidly than the fluorescence based measurements, demonstrating that SECM is suitable for studying the cellular influence of respiration inhibitors. PMID- 12896840 TI - An ultrasensitive and stable potentiometric immunosensor. AB - We describe a novel quantitative polypyrrole based potentiometric biosensor that provides broad-spectrum assay capability. The biosensor allows for capture of analytes of interest from complex real samples such as serum and whole blood, and subsequent measurement in a controlled matrix environment. The technology is rapid (<15 min), ultrasensitive (<50 fM) and reproducible (CV<5% at 0.1 ng/ml). In addition the system has shown a wide dynamic range (four to five orders of magnitude), and good stability, 37 degrees C for at least 4 months. This potentiometric biosensor detects enzyme labelled immuno-complexes formed at the surface of a polypyrrole coated, screenprinted gold electrode. Detection is mediated by a secondary reaction that produces charged products (a 'charge-step' procedure). A shift in potential is measured at the sensor surface, caused by local changes in redox state, pH and/or ionic strength. The magnitude of the difference in potential is related to the concentration of the formed receptor target complex. The potentiometric sensing technology has been demonstrated in assays for hepatitis B surface antigen (HBsAg) (Mw>300 kDa), Troponin I (Mw approximately 23 kDa), Digoxin (Mw 780 Da) and tumour necrosis factor (hTNF alpha) (Mw approximately 23 kDa). These model targets were chosen to represent analytes of a range of molecular weights, and because of their requirement for assays of high analytical sensitivity and precision. All these assays were performed using complex fluid samples and the presence of any non-specific binding has no significant effect on the final measurement. New assays can be transferred and optimised readily. PMID- 12896841 TI - Calibration of the viscometric glucose sensor before its use in physiological liquids--compensation for the colloid-osmotic effect. AB - The function of the recently described viscometric affinity sensor (VAS), which measures glucose due to its strong effect on the viscosity of a sensitive liquid containing Concanavalin A (ConA) and dextran, was analysed for osmotic and colloid-osmotic effects on the glucose reading. The suction of low- and high molecular weight osmotica on the membrane of the microdialysis fibre was measured using a membrane osmometer built from the microdialysis probe of the VAS. The reduction of the sensor read-out in blood plasma can be completely explained by a change in small osmotic volume fluxes through the dialysis membrane, which affect the ConA concentration and the viscosity after the flow of the sensitive liquid through the dialysis probe. The measuring error could be prevented by the presence of the polyethylene glycol 6000 at an isotonic concentration in the glucose standard solutions used for sensor calibration. PMID- 12896842 TI - Sensitivity of the acoustic waveguide biosensor to protein binding as a function of the waveguide properties. AB - The aim of this work is to study the effect of operating frequency, piezoelectric substrate and waveguide layer thickness on the sensitivity of the acoustic waveguide sensor during the specific binding of an antibody by a protein. Shear horizontal (SH) wave devices consisting of (a) a LiTaO3 substrate operating at 104 MHz, (b) a quartz substrate operating at 108 MHz and (c) a quartz substrate operating at 155 MHz were coated with a photoresist polymer layer in order to produce acoustic waveguide devices supporting a Love wave. The effect of the thickness of the polymer layer on the Love wave was assessed by measuring the amplitude and phase of the wave before and after coating. The sensitivity of the above three biosensors was compared during the detection of the specific binding of different concentrations of Immunoglobulin G in the range of 0.7-667 nM to a protein A modified surface. Results indicate that the thickness of the polymer guiding layer is critical for obtaining the maximum sensitivity for a given geometry but a trade-off has to be made between the theoretically determined optimum thickness for waveguiding and the device insertion loss. It was also found that increasing the frequency of operation results in a further increase in the device sensitivity to protein detection. PMID- 12896843 TI - Potential of enzyme mimics in biomimetic sensors: a modified-cyclodextrin as a dehydrogenase enzyme mimic. AB - This paper reports the application of a dehydrogenase enzyme mimic as a biomimetic sensor. The model compound investigated was a beta-cyclodextrin (beta CD) derivative with a nicotinamide group attached to the secondary face of a beta CD (Fig. 1g). It was envisaged that the nicotinamide group would act as the electron transfer agent and that the cyclodextrin would provide a suitable hydrophobic cavity for the reaction to take place in. Ethanol, propranalol, dopamine and acetone were used as substrates in backgrounds of hydrophilic and hydrophobic anions. Electrochemical and fluorescence techniques were used to study the catalytic effects in solution. It was found that the size of the analyte and the hydrophobicity of the anion affected the catalytic activity of the dehydrogenase mimic. Catalytic effects were most enhanced with ethanol and dopamine in presence of larger and more strongly solvated anions, SO4(2-) and H2PO4- which are excluded from the cavity. The molecule was also immobilised in a sol-gel matrix and investigated as a sol-gel electrochemical biomimetic sensor. Concentration dependence with increasing aliquots of ethanol was observed. These results indicated that a re-usable biomimetic sensor is indeed feasible. PMID- 12896844 TI - Real time kinetic analysis of the interaction between immunoglobulin G and histidine using quartz crystal microbalance biosensor in solution. AB - Quartz crystal microbalance (QCM) biosensor integrated in a flow injection analysis (FIA) system was used for the investigation of the specific interaction between immunoglobin G (IgG) and histidine. The histidine was immobilized on the gold electrodes of the piezoelectric crystal using appropriate procedures based on self-assembling of the dithiothreitol (DTT). The specific interaction of the immobilized ligand with IgG in solution was followed as a change in the resonant frequency of the modified crystal and studied in real time without any additional labels. With the mass sensitive biosensor system, the differences in affinity of three different species of IgG: human IgG, goat IgG and mouse IgG were easily distinguished and their respective kinetic rate constants (kass and kdiss) and equilibrium association constants (KA) were determined from the curves of frequency versus time. For the interactions, KA were 2.92 x 10(4), 3.23 x 10(4) and 4.08 x 10(4) M(-1) for human IgG, goat IgG and mouse IgG, respectively. PMID- 12896845 TI - Low-dose exposure to inorganic mercury accelerates disease and mortality in acquired murine lupus. AB - Inorganic mercury (iHg) is known to induce autoimmune disease in susceptible rodent strains. Additionally, in inbred strains of mice prone to autoimmune disease, iHg can accelerate and exacerbate disease manifestations. Despite these well-known links between iHg and autoimmunity in animal models, no association between iHg alone and autoimmune disease in humans has been documented. However, it is possible that low-level iHg exposure can interact with disease triggers to enhance disease expression or susceptibility. To address whether exposure to iHg can alter the course of subsequent acquired autoimmune disease, we used a murine model of acquired autoimmunity, lupus-like chronic graft-versus-host disease (GVHD), in which autoimmunity is induced using normal, nonautoimmune prone donor and F1 recipient mice resistant to Hg-induced autoimmunity. Our results indicate that a 2-week exposure to low-dose iHg (20 or 200 micro g/kg every other day) to donor and host mice ending 1 week before GVHD induction can significantly worsen parameters of disease severity, resulting in premature mortality. iHg pretreatment clearly worsened chronic lupus-like disease, rather than GVHD worsening iHg immunotoxicity. These results are consistent with the hypothesis that low-level, nontoxic iHg preexposure may interact with other risk factors, genetic or acquired, to promote subsequent autoimmune disease development. PMID- 12896847 TI - Exploring bias in a generalized additive model for spatial air pollution data. AB - During the past few years, the generalized additive model (GAM) has become a standard tool for epidemiologic analysis exploring the effect of air pollution on population health. Recently, the use of the GAM has been extended from time series data to spatial data. Still more recently, it has been suggested that the use of GAMs to analyze time-series data results in air pollution risk estimates being biased upward and that concurvity in the time-series data results in standard error estimates being biased downward. We show that concurvity in spatial data can lead to underestimation of the standard error of the estimated air pollution effect, even when using an asymptotically unbiased standard error estimator. We also show that both the magnitude and direction of the bias in the air pollution effect depend, at least in part, on the nature of the concurvity. We argue that including a nonparametric function of location in a GAM for spatial epidemiologic data can be expected to result in concurvity. As a result, we recommend caution in using the GAM to analyze this type of data. PMID- 12896846 TI - Effects of polychlorinated biphenyls on estrogen receptor-beta expression in the anteroventral periventricular nucleus. AB - Polychlorinated biphenyls (PCBs) can disrupt the reproductive axis, particularly when the exposure occurs during the vulnerable developmental periods. Some effects of environmental endocrine disruptors such as PCBs may be exerted through binding to estrogen receptors (ERs). In this study we examined the endocrine disrupting effects of Aroclor 1221 (a commercial PCB mixture), focusing on its actions on the ER-ss, which has been implicated in mediating effects of endocrine disrupting chemicals. A low, ecologically relevant dose of Aroclor 1221 or vehicle (ethanol) was administered three times each to rat dams, on gestational day 16 and on postpartum days 1 and 4, a developmental period during which steroid hormones have permanent effects on adult brain structure and function. Effects on ER-ss cell number in the anteroventral periventricular nucleus (AVPV) were quantified; this sexually dimorphic nucleus of the brain is essential to female reproductive function. For comparison, we quantified ER-ss cell number in another hypothalamic region, the supraoptic nucleus (SON). Using a stereologic approach, we found that Aroclor 1221 caused a highly significant down-regulation of the number of ER-ss-expressing cells in the AVPV, but had no effect in the SON. Thus, PCB exposure has consequences for neural ER expression, and these findings have implications for wildlife and humans that have been exposed to environmental estrogens, particularly during the susceptible periods of early development. PMID- 12896848 TI - Proinflammatory and cytotoxic effects of Mexico City air pollution particulate matter in vitro are dependent on particle size and composition. AB - Exposure to urban airborne particulate matter (PM) is associated with adverse health effects. We previously reported that the cytotoxic and proinflammatory effects of Mexico City PM10 (less than or equal to 10 micro m mean aerodynamic diameter) are determined by transition metals and endotoxins associated with these particles. However, PM2.5 (less than or equal to 2.5 micro m mean aerodynamic diameter) could be more important as a human health risk because this smaller PM has the potential to reach the distal lung after inhalation. In this study, we compared the cytotoxic and proinflammatory effects of Mexico City PM10 with those of PM2.5 using the murine monocytic J774A.1 cell line in vitro. PMs were collected from the northern zone or the southeastern zone of Mexico City. Elemental composition and bacterial endotoxin on PMs were measured. Tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) production by J774A.1 cells was measured in the presence or absence of recombinant endotoxin neutralizing protein (rENP). Both northern and southeastern PMs contained endotoxin and a variety of transition metals. Southeastern PM10 contained the highest endotoxin levels, 2-fold higher than that in northern PM10. Northern and southeastern PM2.5 contained the lowest endotoxin levels. Accordingly, southeastern PM10 was the most potent in causing secretion of the proinflammatory cytokines TNF-alpha and IL-6. All PM2.5 and PM10 samples caused cytotoxicity, but northern PMs were the most toxic. Cytokine secretion induced by southeastern PM10 was reduced 50-75% by rENP. These results indicate major differences in PM10 and PM2.5. PM2.5 induces cytotoxicity in vitro through an endotoxin-independent mechanism that is likely mediated by transition metals. In contrast, PM10 with relatively high levels of endotoxin induces proinflammatory cytokine release via an endotoxin-dependent mechanism. PMID- 12896849 TI - Diminished experience-dependent neuroanatomical plasticity: evidence for an improved biomarker of subtle neurotoxic damage to the developing rat brain. AB - Millions of children are exposed to low levels of environmental neurotoxicants as their brains are developing. Conventional laboratory methods of neurotoxicology can detect maldevelopment of brain structure but are not designed to detect maldevelopment of the brain's capacity for plasticity that could impair learning throughout life. The environmental complexity (EC) paradigm has become classic for demonstrating the modifications in brain structure that occur in response to experience and thus provides a set of indices for plasticity in the healthy brain. In this study, we have tested the hypothesis that if degradation of experience-dependent cortical plasticity is used as a biomarker, then developmental neurotoxic effects will be detected at doses below those that alter cortical morphogenesis overtly. Pregnant Long-Evans hooded rats received a single injection of either saline vehicle or 1, 5, 10, or 25 mg/kg of the well characterized developmental neurotoxicant methylazoxymethanol acetate (MAM) on the 16th or 17th day of gestation. On postnatal days 35-39, male offspring were assigned to either a complex environment (EC) or an individual cage (IC) for 28 days to stimulate neuroanatomical plasticity. This response was measured as the difference between the thickness of visual cortex of IC and EC littermates at a given dose. The threshold dose for significant reduction of cortical thickness was 25 mg/kg, but the threshold dose for failure of plasticity was much lower and could be detected at 1 mg/kg, the lowest dose used. No other method of assessment has detected lasting effects of prenatal exposure to MAM at such a low dose. These data suggest that this simple test of plasticity could be an efficient way to detect subtle neurotoxic damage to the developing brain. PMID- 12896850 TI - Aryl hydrocarbon receptor-mediated activity of particulate organic matter from the Paso del Norte airshed along the U.S.-Mexico border. AB - In this study, we determined the biologic activity of dichloromethane-extracted particulate matter < 10 micro m in aerodynamic diameter (PM10) obtained from filters at three sites in the Paso del Norte airshed, which includes El Paso, Texas, USA; Juarez, Chihuahua, Mexico, and Sunland Park, New Mexico, USA. The extracts were rich in polycyclic aromatic hydrocarbons (PAHs) and had significant biologic activity, measured using two in vitro assay systems: ethoxyresorufin-(O deethylase (EROD) induction and the aryl hydrocarbon-receptor luciferase reporter system. In most cases, both EROD (5.25 pmol/min/mg protein) and luciferase activities (994 relative light units/mg) were highest in extracts from the Advance site located in an industrial neighborhood in Juarez. These values represented 58% and 55%, respectively, of induction associated with 1 micro M ss naphthoflavone exposures. In contrast, little activity was observed at the Northeast Clinic site in El Paso, the reference site. In most cases, luciferase and EROD activity from extracts collected from the Tillman Health Center site, situated in downtown El Paso, fell between those observed at the other two sites. Overall, a statistically significant correlation existed between PM10 and EROD and luciferase activities. Chemical analysis of extracts collected from the Advance site demonstrated that concentrations of most PAHs were higher than those reported in most other metropolitan areas in the United States. Calculations made with these data suggest a cancer risk of 5-12 cases per 100,000 people. This risk estimate, as well as comparisons with the work of other investigators, raises concern regarding the potential for adverse health effects to the residents of this airshed. Further work is needed to understand the sources, exposure, and effects of PM10 and particulate organic material in the Paso del Norte airshed. PMID- 12896851 TI - Assessment of dietary exposure to some persistent organic pollutants in the Republic of Karakalpakstan of Uzbekistan. AB - A 1999 study heightened long-standing concerns over persistent organic pollutant contamination in the Aral Sea area, detecting elevated levels in breast milk and cord blood of women in Karakalpakstan (western Uzbekistan). These findings prompted a collaborative research study aimed at linking such human findings with evidence of food chain contamination in the area. An international team carried out analyses of organochlorine and organophosphate pesticides, polychlorinated biphenyls (PCBs), polychlorinated dibenzo-p-dioxins (PCDDs), and polychlorinated dibenzofurans (PCDFs) on samples of 12 foods commonly produced and consumed in Karakalpakstan. Analysis consistently detected long-lasting organochlorine pesticides and their metabolites in all foods of animal origin and in some vegetables such as onions and carrots--two low-cost components of many traditional dishes. Levels of PCBs were relatively low in all samples except fish. Analyses revealed high levels of PCDDs and PCDFs (together often termed "dioxins") in sheep fat, dairy cream, eggs, and edible cottonseed oil, among other foodstuffs. These findings indicate that food traditionally grown, sold, and consumed in Karakalpakstan is a major route of human exposure to several persistent toxic contaminants, including the most toxic of dioxins, 2,3,7,8 tetrachlorodibenzo-p-dioxin (2,3,7,8-TCDD). Intake estimations demonstrate that consumption of even small amounts of locally grown food may expose consumers to dioxin levels that considerably exceed the monthly tolerable dioxin intake levels set by the World Health Organization. Data presented in this study allow a first assessment of the risk associated with the consumption of certain food products in Karakalpakstan and highlight a critical public health situation. PMID- 12896853 TI - Assessment of potential risk levels associated with U.S. Environmental Protection Agency reference values. AB - The U.S. Environmental Protection Agency (U.S. EPA) generally uses reference doses (RfDs) or reference concentrations (RfCs) to assess risks from exposure to toxic substances for noncancer health end points. RfDs and RfCs are supposed to represent lifetime inhalation or ingestion exposure with minimal appreciable risk, but they do not include information about the estimated risk from exposures equal to the RfD/RfC. We used results from benchmark dose modeling approaches recently adopted for use in developing RfDs/RfCs to estimate the risk levels associated with exposures at the RfD/RfC. We searched the U.S. EPA Integrated Risk Information System (IRIS) database and identified 11 chemicals with oral RfDs and 12 chemicals with inhalation RfCs that used benchmark dose modeling. For assessments with sufficient model information, we found that 16 of 21 (76%) of the dose-response models were linear or supralinear. We estimated the risk from exposures at the established RfDs and RfCs for these chemicals using a linear dose-response curve to characterize risk below the observed data. Risk estimates ranged from 1 in 10,000 to 5 in 1,000 for exposures at the RfDs, and from 1 in 10,000 to 3 in 1,000 for exposures at the RfCs. Risk estimates for exposures at the RfD/RfC values derived from sublinear dose-response curves ranged from 3 in 1,000,000,000 to 8 in 10,000. Twenty-four percent of reference values corresponded to estimated risk levels greater than 1 in 1,000; 10 of 14 assessments had points of departure greater than the no-observed-adverse-effect levels. For policy development regarding management of cancer risks, the U.S. EPA often uses 1 in 1,000,000 as a de minimis risk level. Although noncancer outcomes may in some instances be reversible and considered less severe than cancer, our findings call into question the assumption that established RfD and RfC values represent negligibly small risk levels. PMID- 12896852 TI - Temperature, air pollution, and hospitalization for cardiovascular diseases among elderly people in Denver. AB - Daily measures of maximum temperature, particulate matter less than or equal to 10 micro m in aerodynamic diameter (PM10), and gaseous pollution (ozone, nitrogen dioxide, sulfur dioxide, and carbon monoxide) were collected in Denver, Colorado, in July and August between 1993 and 1997. We then compared these exposures with concurrent data on the number of daily hospital admissions for cardiovascular diseases in men and women > 65 years of age. Generalized linear models, assuming a Poisson error structure for the selected cardiovascular disease hospital admissions, were constructed to evaluate the associations with air pollution and temperature. After adjusting the admission data for yearly trends, day-of-week effects, ambient maximum temperature, and dew point temperature, we studied the associations of the pollutants in single-pollutant models with lag times of 0-4 days. The results suggest that O3 is associated with an increase in the risk of hospitalization for acute myocardial infarction, coronary atherosclerosis, and pulmonary heart disease. SO2 appears to be related to increased hospital stays for cardiac dysrhythmias, and CO is significantly associated with congestive heart failure. No association was found between particulate matter or NO2 and any of the health outcomes. Males tend to have higher numbers of hospital admissions than do females for all of the selected cardiovascular diseases, except for congestive heart failure. Higher temperatures appear to be an important factor in increasing the frequency of hospitalization for acute myocardial infarction and congestive heart failure, and are associated with a decrease in the frequency of visits for coronary atherosclerosis and pulmonary heart disease. PMID- 12896854 TI - Neurologic outcomes of toxic oil syndrome patients 18 years after the epidemic. AB - Toxic oil syndrome (TOS) resulted from consumption of rapeseed oil denatured with 2% aniline and affected more than 20,000 persons. Eighteen years after the epidemic, many patients continue to report neurologic symptoms that are difficult to evaluate using conventional techniques. We conducted an epidemiologic study to determine whether an exposure to toxic oil 18 years ago was associated with current adverse neurobehavioral effects. We studied a case group of 80 adults exposed to toxic oil 18 years ago and a referent group of 79 adult age- and sex frequency-matched unexposed subjects. We interviewed subjects for demographics, health status, exposures to neurotoxicants, and responses to the Kaufman Brief Intelligence Test (K-BIT), Programa Integrado de Exploracion Neuropsicologica (PIEN), and Goldberg depression questionnaires and administered quantitative neurobehavioral and neurophysiologic tests by computer or trained nurses. The groups did not differ with respect to educational background or other critical variables. We examined associations between case and referent groups and the neurobehavioral and neurophysiologic outcomes of interest. Decreased distal strength of the dominant and nondominant hands and increased vibrotactile thresholds of the fingers and toes were significantly associated with exposure to toxic oil. Finger tapping, simple reaction time latency, sequence B latency, symbol digit latency, and auditory digit span were also significantly associated with exposure. Case subjects also had statistically significantly more neuropsychologic symptoms compared with referents. Using quantitative neurologic tests, we found significant adverse central and peripheral neurologic effects in a group of TOS patients 18 years after exposure to toxic oil when compared with a nonexposed referent group. These effects were not documented by standard clinical examination and were found more frequently in women. PMID- 12896855 TI - Amyotrophic lateral sclerosis, lead, and genetic susceptibility: polymorphisms in the delta-aminolevulinic acid dehydratase and vitamin D receptor genes. AB - Previous studies have suggested that lead exposure may be associated with increased risk of amyotrophic lateral sclerosis (ALS). Polymorphisms in the genes for delta-aminolevulinic acid dehydratase (ALAD) and the vitamin D receptor (VDR) may affect susceptibility to lead exposure. We used data from a case-control study conducted in New England from 1993 to 1996 to evaluate the relationship of ALS to polymorphisms in ALAD and VDR and the effect of these polymorphisms on the association of ALS with lead exposure. The ALAD 2 allele (177G to C; K59N) was associated with decreased lead levels in both patella and tibia, although not in blood, and with an imprecise increase in ALS risk [odds ratio (OR) = 1.9; 95% confidence interval (95% CI), 0.60-6.3]. We found a previously unreported polymorphism in ALAD at an Msp1 site in intron 2 (IVS2+299G>A) that was associated with decreased bone lead levels and with an imprecise decrease in ALS risk (OR = 0.35; 95% CI, 0.10-1.2). The VDR B allele was not associated with lead levels or ALS risk. Our ability to observe effects of genotype on associations of ALS with occupational exposure to lead or with blood or bone lead levels was limited. These findings suggest that genetic susceptibility conferred by polymorphisms in ALAD may affect ALS risk, possibly through a mechanism related to internal lead exposure. PMID- 12896856 TI - Environmental threats to children's health in Southeast Asia and the Western Pacific. AB - The Southeast Asia and Western Pacific regions contain half of the world's children and are among the most rapidly industrializing regions of the globe. Environmental threats to children's health are widespread and are multiplying as nations in the area undergo industrial development and pass through the epidemiologic transition. These environmental hazards range from traditional threats such as bacterial contamination of drinking water and wood smoke in poorly ventilated dwellings to more recently introduced chemical threats such as asbestos construction materials; arsenic in groundwater; methyl isocyanate in Bhopal, India; untreated manufacturing wastes released to landfills; chlorinated hydrocarbon and organophosphorous pesticides; and atmospheric lead emissions from the combustion of leaded gasoline. To address these problems, pediatricians, environmental health scientists, and public health workers throughout Southeast Asia and the Western Pacific have begun to build local and national research and prevention programs in children's environmental health. Successes have been achieved as a result of these efforts: A cost-effective system for producing safe drinking water at the village level has been devised in India; many nations have launched aggressive antismoking campaigns; and Thailand, the Philippines, India, and Pakistan have all begun to reduce their use of lead in gasoline, with resultant declines in children's blood lead levels. The International Conference on Environmental Threats to the Health of Children, held in Bangkok, Thailand, in March 2002, brought together more than 300 representatives from 35 countries and organizations to increase awareness on environmental health hazards affecting children in these regions and throughout the world. The conference, a direct result of the Environmental Threats to the Health of Children meeting held in Manila in April 2000, provided participants with the latest scientific data on children's vulnerability to environmental hazards and models for future policy and public health discussions on ways to improve children's health. The Bangkok Statement, a pledge resulting from the conference proceedings, is an important first step in creating a global alliance committed to developing active and innovative national and international networks to promote and protect children's environmental health. PMID- 12896857 TI - Distribution and determinants of mouse allergen exposure in low-income New York City apartments. AB - Previous studies of mouse allergens and laboratory-animal-worker-related allergy and asthma suggest that quantifying mouse allergen levels in homes could augment our understanding of inner-city asthma. We hypothesized that levels of mouse allergen in inner-city homes would be related to certain household characteristics. Dust samples were collected from the kitchens and beds of 221 mothers enrolled in a prospective birth cohort study, 92 of African American and 129 of Dominican ethnicity. Samples were analyzed for mouse urinary protein. The geometric mean for kitchen samples was 4.6 micro g/g [95% confidence interval (95% CI), 3.2-6.5] and for bed samples was 0.9 micro g/g (95% CI, 0.8-1.1). The variables associated with mouse allergen levels in the home were frequency of mouse sightings, use of traps or pesticides for mice, presence of holes in ceilings or walls, absence of a cat, and living in a building with fewer than eight floors. Statistically significant neighborhood differences in levels of mouse allergen and report of rodents in the home were also observed. In conclusion, mouse allergen was prevalent among inner-city apartments, and the positive predictive value of self-reported frequent mouse sightings was high (90% for kitchens). However, high levels of mouse allergen were also found in many homes where mothers reported never seeing mice. PMID- 12896858 TI - Low birth weight and residential proximity to PCB-contaminated waste sites. AB - Previous investigations have shown that women exposed to polychlorinated biphenyls (PCBs) are at increased risk of giving birth to an infant with low birth weight (< 2,500 g), and that this relationship is stronger for male than for female infants. We have tested the hypothesis that residents in a zip code that contains a PCB hazardous waste site or abuts a body of water contaminated with PCBs are at increased risk of giving birth to a low-birth-weight baby. We used the birth registry of the New York State Vital Statistics to identify all births between 1994 and 2000 in New York State except for New York City. This registry provides information on the infant, mother, and father together with the zip code of the mother's residence. The 865 state Superfund sites, the 86 National Priority List sites, and the six Areas of Concern in New York were characterized regarding whether or not they contain PCBs as a major contaminant. We identified 187 zip codes containing or abutting PCB-contaminated sites, and these zip codes were the residences of 24.5% of the 945,077 births. The birth weight in the PCB zip codes was on average 21.6 g less than in other zip codes (p < 0.001). Because there are many other risk factors for low birth weight, we have adjusted for these using a logistic regression model for these confounders. After adjusting for sex of the infant, mother's age, race, weight, height, education, income, marital status, and smoking, there was still a statistically significant 6% increased risk of giving birth to a male infant of low birth weight. These observations support the hypothesis that living in a zip code near a PCB contaminated site poses a risk of exposure and giving birth to an infant of low birth weight. PMID- 12896859 TI - Summary of a workshop on regulatory acceptance of (Q)SARs for human health and environmental endpoints. AB - The "Workshop on Regulatory Use of (Q)SARs for Human Health and Environmental Endpoints," organized by the European Chemical Industry Council and the International Council of Chemical Associations, gathered more than 60 human health and environmental experts from industry, academia, and regulatory agencies from around the world. They agreed, especially industry and regulatory authorities, that the workshop initiated great potential for the further development and use of predictive models, that is, quantitative structure activity relationships [(Q)SARs], for chemicals management in a much broader scope than is currently the case. To increase confidence in (Q)SAR predictions and minimization of their misuse, the workshop aimed to develop proposals for guidance and acceptability criteria. The workshop also described the broad outline of a system that would apply that guidance and acceptability criteria to a (Q)SAR when used for chemical management purposes, including priority setting, risk assessment, and classification and labeling. PMID- 12896860 TI - Methods for reliability and uncertainty assessment and for applicability evaluations of classification- and regression-based QSARs. AB - This article provides an overview of methods for reliability assessment of quantitative structure-activity relationship (QSAR) models in the context of regulatory acceptance of human health and environmental QSARs. Useful diagnostic tools and data analytical approaches are highlighted and exemplified. Particular emphasis is given to the question of how to define the applicability borders of a QSAR and how to estimate parameter and prediction uncertainty. The article ends with a discussion regarding QSAR acceptability criteria. This discussion contains a list of recommended acceptability criteria, and we give reference values for important QSAR performance statistics. Finally, we emphasize that rigorous and independent validation of QSARs is an essential step toward their regulatory acceptance and implementation. PMID- 12896861 TI - Use of QSARs in international decision-making frameworks to predict ecologic effects and environmental fate of chemical substances. AB - This article is a review of the use, by regulatory agencies and authorities, of quantitative structure-activity relationships (QSARs) to predict ecologic effects and environmental fate of chemicals. For many years, the U.S. Environmental Protection Agency has been the most prominent regulatory agency using QSARs to predict the ecologic effects and environmental fate of chemicals. However, as increasing numbers of standard QSAR methods are developed and validated to predict ecologic effects and environmental fate of chemicals, it is anticipated that more regulatory agencies and authorities will find them to be acceptable alternatives to chemical testing. PMID- 12896862 TI - Use of QSARs in international decision-making frameworks to predict health effects of chemical substances. AB - This article is a review of the use of quantitative (and qualitative) structure activity relationships (QSARs and SARs) by regulatory agencies and authorities to predict acute toxicity, mutagenicity, carcinogenicity, and other health effects. A number of SAR and QSAR applications, by regulatory agencies and authorities, are reviewed. These include the use of simple QSAR analyses, as well as the use of multivariate QSARs, and a number of different expert system approaches. PMID- 12896863 TI - Emerging diseases threaten conservation. PMID- 12896864 TI - Re: analysis of dioxin cancer threshold. PMID- 12896865 TI - Exposure to lead and an old way of counting. PMID- 12896866 TI - Carbon monoxide exposure and carboxyhemoglobin. PMID- 12896868 TI - Mountain tourism: news from Nepal. PMID- 12896869 TI - Health hazards and healthcare costs. PMID- 12896870 TI - Conservation medicine: combining the best of all worlds. PMID- 12896871 TI - Subjective science: environmental cost-benefit analysis. PMID- 12896872 TI - Solvent shakedown: the mechanochemistry alternative. PMID- 12896874 TI - Heterogeneity of heparan sulfates in human lung. AB - Heparan sulfates (HS), a class of glycosaminoglycans, are long linear complex polysaccharides covalently attached to a protein core. The HS molecules are made up of repeating disaccharides onto which modification patterns are superimposed. This results in a large structural heterogeneity and forms the basis of specific interactions of HS toward a vast array of proteins, including growth factors and proteases. To study HS heterogeneity in the lung, we used phage display technology to select seven antibodies against human lung HS. Antibodies reacted with HS/heparin, but not with other glycosaminoglycans or polyanions. Sulfate groups were essential for antibody binding. The amino acid sequence of the antibodies was established, the complementarity determining region 3 of the heavy chain containing basic amino acids. The antibodies defined HS epitopes with a characteristic tissue distribution. Antibody EV3A1 primarily stained macrophages. Other antibodies primarily stained basement membranes, but with different preference toward type of basement membrane. Antibody EV3C3 was the only antibody which clearly reacted with bronchiolar epithelial cells. In human lung parenchyma, basic fibroblast growth factor and vascular endothelial growth factor were largely bound by HS. Some antibodies blocked a basic fibroblast growth factor-binding site of HS, and one antibody blocked a vascular endothelial growth factor-binding site of heparin. Taken together, these data suggest a specific role for HS epitopes in human lung. The antibodies obtained may be valuable tools to study HS in pulmonary diseases. PMID- 12896875 TI - Lipogenesis in fetal rat lung: importance of C/EBPalpha, SREBP-1c, and stearoyl CoA desaturase. AB - Alveolar type II cells increase lipogenesis and convert glycogen into the phospholipids of surfactant in the late term fetal lung. Recent studies suggest that CCAAT/enhancing-binding protein (C/EBP) isoforms and sterol regulatory element binding protein (SREBP)-1c regulate fatty acid synthesis in adult type II cells in vitro. To define the temporal relationships and enzymes involved in lipogenesis in fetal rat lung, the mRNA levels of selected transcription factors and enzymes were determined. There was an increase in the mRNA levels of C/EBPalpha, C/EBPbeta, C/EBPdelta, peroxisomal proliferator-activated receptor gamma (PPARgamma), and SREBP-1c, but not SREBP-1a or SREBP-2 from fetal Days 19 21. There was also an increase in the mRNA levels of fatty acid synthase, stearoyl-CoA desaturase 1 (SCD-1), fatty acid translocase, glycerol-3-P acyl transferase, and phosphatidate cytidylyltransferase. By in situ hybridization, there was detectible expression of fatty acid synthase, SCD-1, and C/EBPalpha along the alveolar septae with the same distribution pattern as surfactant protein-C, whereas PPARgamma expression appeared to be restricted to macrophages. Regulation of lipogenesis at the mRNA level is predominately on enzymes of fatty acid synthesis and appears to be regulated by C/EBPalpha and SREBP-1c. SCD-1 and phosphatidate cytidylyltransferase are important components of the lipogenic response in the fetal lung that have not been recognized previously. PMID- 12896876 TI - Disrupted pulmonary vascular development and pulmonary hypertension in transgenic mice overexpressing transforming growth factor-alpha. AB - Pulmonary vascular disease plays a major role in morbidity and mortality in infant and adult lung diseases in which increased levels of transforming growth factor (TGF)-alpha and its receptor EGFR have been associated. The aim of this study was to determine whether overexpression of TGF-alpha disrupts pulmonary vascular development and causes pulmonary hypertension. Lung-specific expression of TGF-alpha in transgenic mice was driven with the human surfactant protein (SP) C promoter. Pulmonary arteriograms and arterial counts show that pulmonary vascular development was severely disrupted in TGF-alpha mice. TGF-alpha mice developed severe pulmonary hypertension and vascular remodeling characterized by abnormally extensive muscularization of small pulmonary arteries. Pulmonary vascular development was significantly improved and pulmonary hypertension and vascular remodeling were prevented in bi-transgenic mice expressing both TGF alpha and a dominant-negative mutant EGF receptor under the control of the SP-C promoter. Vascular endothelial growth factor (VEGF-A), an important angiogenic factor produced by the distal epithelium, was decreased in the lungs of TGF-alpha adults and in the lungs of infant TGF-alpha mice before detectable abnormalities in pulmonary vascular development. Hence, overexpression of TGF-alpha caused severe pulmonary vascular disease, which was mediated through EGFR signaling in distal epithelial cells. Reductions in VEGF may contribute to the pathogenesis of pulmonary vascular disease in TGF-alpha mice. PMID- 12896877 TI - Nitric oxide inhibits surfactant protein B gene expression in lung epithelial cells. AB - Surfactant protein B (SP-B) is an essential constituent of pulmonary surfactant. In a number of inflammatory diseases of the lung, elevated nitric oxide (NO) levels are associated with decreased SP-B levels, suggesting that reduced SP-B levels contribute to lung injury. In this study, we investigated the effects of NO on SP-B gene expression in H441 and MLE-12 cells, cell lines with characteristics of bronchiolar (Clara) and alveolar type II epithelial cells, respectively. Results show that NO donors decreased SP-B mRNA levels in a concentration- and time-dependent manner in H441 and MLE-12 cells. The NO donors also antagonized dexamethasone induction of SP-B mRNA in H441 cells. NO donor inhibition of SP-B mRNA was blocked by the transcriptional inhibitor 5,6-dichloro 1-beta-D-ribofuranozyl-benzimidazole. NO donors decreased luciferase expression from a reporter plasmid containing -911/+41 bp of human SP-B 5'-flanking DNA in H441 and MLE-12 cells, indicating inhibitory effects on SP-B promoter activity. NO inhibition of SP-B mRNA levels was not blocked by LY-83583 and KT-5823, inhibitors of soluble guanylate cyclase and protein kinase G, respectively. Furthermore, cell-permeable cGMP analog 8-bromo-cGMP had no effect on SP-B mRNA levels. These data indicate that elevated NO levels negatively regulate SP-B gene expression by inhibiting gene transcription and that NO inhibits SP-B gene expression independently of cGMP levels. These data imply that reduced SP-B expression due to elevated NO levels can contribute to lung injury. PMID- 12896878 TI - Low-dose carbon monoxide reduces airway hyperresponsiveness in mice. AB - Carbon monoxide (CO) in expired gas has been shown to be elevated with asthma; however, its function is not known, and there is some potential that it may serve a bronchoprotective role to decrease airway hyperresponsiveness (AHR). Thus the ability of CO to reverse methacholine (MCh)-induced bronchoconstriction was evaluated in C57BL/6 (C57) and A/J mice with and without airway inflammation produced by ovalbumin (OVA). Acutely administered CO (1% in air, 10 min) reduced MCh-driven increases in lung resistance in OVA-challenged C57 mice by an average of 50% (from 14.5 to 7.1 cmH2O.ml-1.s-1), whereas no effect was observed in naive C57 mice or OVA-challenged C57 mice inhaling air alone. Acutely inhaled CO (500 ppm = 0.05%, for 10 min) reduced MCh-induced airway reactivity (AR) by 20-60% in airway hyperresponsive naive A/J mice, whereas repeated 10-min administrations of 500 ppm CO over a 5-day period decreased AR by 50%. Repeated administration of low-dose CO [250 (0.025%) and (0.05%) 500 ppm, 1 h/day, 5 days] to A/J mice with airway inflammation likewise resulted in a drop of AR by 50%, compared with those not receiving CO. Inhibition of guanylyl cyclase/guanosine 3',5'-cyclic monophosphothioate (cGMP) using 1H-[1,2,4] oxydiazolo[4,3-a]quinoxalin-1-one or a competitive inhibitor, Rp diastereomers of 8-bromo-cGMP, resulted in inhibition of the effect of CO on AHR, suggesting that the effects of CO were mediated through this mechanism. These results indicate that low-dose CO can effectively reverse AHR in the presence and absence of airway inflammation in mice and suggest a potential role for CO in the modulation of AHR. PMID- 12896879 TI - Increased iNOS activity is essential for pulmonary epithelial tight junction dysfunction in endotoxemic mice. AB - A murine endotoxemia model and cultured Calu-3 monolayers were used to test the hypothesis that excessive nitric oxide (NO) production secondary to induction of inducible NO synthase (iNOS) is a key factor leading to altered tight junction (TJ) protein expression and function in the pulmonary epithelium. C57Bl/6J mice were injected with either Escherichia coli 0111:B4 lipopolysaccharide (LPS; 2 mg/kg) or vehicle. Twelve hours later, leakage of FITC-dextran (M(r) 4 kDa; FD4) from blood into bronchoalveolar lavage fluid was significantly increased in endotoxemic but not control mice. This decrease in bronchoalveolar barrier function was associated with upregulation of iNOS protein expression and NF kappaB activation in lung tissue. Expression of the TJ proteins, zonula occludens (ZO)-1, ZO-2, ZO-3, and occludin, as assessed by immunoblotting and/or immunofluorescence, decreased in lung after the injection of mice with LPS. Treatment of endotoxemic mice with an isoform-selective iNOS inhibitor [l-N(6)-(1 iminoethyl)lysine; l-NIL] ameliorated LPS-induced changes in TJ protein expression and preserved bronchoalveolar epithelial barrier function. Incubating Calu-3 bronchiolar epithelial monolayers with cytomix (a mixture of 1,000 U/ml IFN-gamma, 10 ng/ml TNF-alpha, and 1 ng/ml IL-1beta) increased permeability to FD4, but adding l-NIL prevented this effect. These results suggest that decreased expression and mistargeting of TJ proteins in lung after systemic inflammation may be NO dependent. PMID- 12896880 TI - PU.1 regulation of human alveolar macrophage differentiation requires granulocyte macrophage colony-stimulating factor. AB - Granulocyte-macrophage colony-stimulating factor (GM-CSF) is critically implicated in lung homeostasis in the GM-CSF knockout mouse model. These animals develop an isolated lung lesion reminiscent of pulmonary alveolar proteinosis (PAP) seen in humans. The development of the adult form of human alveolar proteinosis is not due to the absence of a GM-CSF gene or receptor defect but to the development of an anti-GM-CSF autoimmunity. The role of GM-CSF in the development of PAP is unknown. Studies in the GM-CSF knockout mouse have shown that lack of PU.1 protein expression in alveolar macrophages is correlated with decreased maturation, differentiation, and surfactant catabolism. This study investigates PU.1 expression in vitro and in vivo in human PAP alveolar macrophages as well as the regulation of PU.1 by GM-CSF. We show for the first time that PU.1 mRNA expression in PAP bronchoalveolar lavage cells is deficient compared with healthy controls. PU.1-dependent terminal differentiation markers CD32 (FCgammaII), mannose receptor, and macrophage colony-stimulating factor receptor (M-CSFR) are decreased in PAP alveolar macrophages. In vitro studies demonstrate that exogenous GMCSF treatment upregulated PU.1 and M-CSFR gene expression in PAP alveolar macrophages. Finally, in vivo studies showed that PAP patients treated with GM-CSF therapy have higher levels of PU.1 and M-CSFR expression in alveolar macrophages compared with healthy control and PAP patients before GM-CSF therapy. These observations suggest that PU.1 is critical in the terminal differentiation of human alveolar macrophages. PMID- 12896881 TI - Impact of helical computed tomography on the outcomes of emergency department patients with suspected appendicitis. AB - OBJECTIVES: To assess the impact of an emergency department (ED) guideline employing selective use of helical computed tomography (CT) on clinical outcomes of female patients with suspected appendicitis. METHODS: All patients presenting with suspected appendicitis were prospectively enrolled and managed in accordance with a guideline incorporating selective use of helical CT. Although not the objective of this investigation, male patients were included for purposes of comparison. Patients with clinically evident appendicitis were referred to the surgical service, and patients with equivocal presentations were studied with helical CT. Patients were followed to final surgical or clinical outcomes. Outcome measures included time from ED presentation to laparotomy and rate of appendiceal perforation. These measures were compared with those of a historical cohort of patients preceding the use of helical CT. RESULTS: A total of 310 consecutive patients with suspected appendicitis were enrolled; 92 had appendicitis. Sixty patients were referred to the surgical service without helical CT, and 41 had appendicitis (68%). Helical CT was performed on 250 patients; 51 had appendicitis (20%). For males, the mean interval from ED presentation to laparotomy was 559 minutes (95% CI = 444 to 674 minutes) during guideline use and 480 minutes (95% CI = 405 to 555 minutes) before. This interval for females was 433 minutes (95% CI = 326 to 540 minutes) during guideline use and 710 minutes (95% CI = 558 to 862 minutes) before. Appendiceal perforation rate for males was 0.25 (95% CI = 0.14 to 0.36) during guideline use and 0.38 (95% CI = 0.29 to 0.47) before; perforation rate for females was 0.06 (95% CI = 0.05 to 0.17) during guideline use and 0.23 (95% CI = 0.14 to 0.32) before. Helical CT had 92% sensitivity, 97% specificity, and 96% accuracy in diagnosing appendicitis. CONCLUSIONS: Helical CT is highly accurate in detecting appendicitis in patients with equivocal ED presentations. The use of a guideline employing selective helical CT was associated with a decline in the time from ED presentation to operative intervention in females. PMID- 12896882 TI - Interrater reliability of criteria used in assessing blunt head injury patients for intracranial injuries. AB - OBJECTIVE: To determine the interrater reliability of potential predictor variables that may be used to construct a clinical decision rule for emergency computed tomography of the head in blunt head injury victims. METHODS: As a substudy of the NEXUS II study of blunt head trauma, physicians from 21 different emergency departments performed paired evaluations of patients undergoing computed tomography of the head after blunt head injury. Each physician independently determined, for each subject, the presence or absence of each of 19 separate clinical characteristics. The physicians were either residents or attending physicians in the participating emergency departments. Paired responses on a sample of 3,951 patients were compared for raw level of agreement and for interrater concordance using the kappa statistic. If the lower margin of the 95% confidence interval for raw agreement was at least 85% and was equal to or greater than 0.50 for kappa, this was predetermined to represent substantial interrater agreement. RESULTS: There was substantial interobserver agreement by both measures for 17 of the 19 candidate variables in patients with blunt head trauma. Interobserver agreement was substantial for all candidate variables except presence of seizure (kappa = 0.57 [95% CI = 0.47 to 0.67]; raw agreement = 96.5%) and abnormal cerebellar function (kappa = 0.54 [95% CI = 0.41 to 0.67]; raw agreement = 96.5%). CONCLUSIONS: The clinicians in our study had a substantial level of agreement regarding most clinical criteria assessed in this large sample of patients with blunt head injury. PMID- 12896883 TI - Arterial blood gas results rarely influence emergency physician management of patients with suspected diabetic ketoacidosis. AB - OBJECTIVES: To test the hypothesis that arterial blood gas (ABG) results for patients with suspected diabetic ketoacidosis (DKA) do not influence emergency physicians' management decisions and to assess correlation and precision between venous pH and arterial pH. METHODS: Prospective, observational study of emergency physicians' decision making for consecutive ED patients with suspected DKA. Inclusion criteria were capillary blood glucose equal to or greater than 200 mg/dL, ketonuria, and clinical signs and symptoms of DKA. Venous pH, chemistry panel, and ABGs were drawn before treatment. Attending emergency physicians indicated planned management and disposition on a standardized form before and after reviewing ABG and venous pH results. This study was powered to detect a 10% difference in management decisions (n = 195). Pearson's correlation and Bland Altman bias plot were used to compare venous pH and arterial pH. RESULTS: ABG analysis changed the emergency physicians' diagnosis in 2/200 cases (1.0%; 95% confidence interval [95% CI] = 0.3% to 3.6%), altered treatment in 7/200 cases (3.5%; 95% CI = 1.7% to 7.1%), and changed disposition in 2/200 cases (1.0%; 95% CI = 0.3% to 3.6%). The pH value of the ABGs changed the treatment or disposition in 5/200 patients (2.5%; 95% CI = 1.1% to 5.7%). The Po(2) and Pco(2) results of the ABGs altered treatment and disposition in 2/200 patients (1.0%; 95% CI = 0.3% to 3.6%). Venous pH correlated well with arterial pH (r = 0.951), and bias plotting yielded a bias value of -0.015 (+/- 0.006 pH units). CONCLUSIONS: ABG results rarely influenced emergency physicians' decisions on diagnosis, treatment, or disposition in suspected DKA patients. Venous pH correlated well and was precise enough with arterial pH to serve as a substitute. PMID- 12896884 TI - Efficient communication: assessment-oriented oral case presentation. AB - OBJECTIVE: To introduce and assess the time savings from and effectiveness of assessment-oriented (AO) oral case presentation as a model of interphysician communication. METHODS: This was a prospective, interventional study of all 10 on site faculty and 36 residents in a postgraduate year 1 to 3 format emergency medicine residency training program. Residents were requested to perform all oral case presentations in either the traditional or AO formats. Presentations were timed, and residents and faculty rated essential measures of oral case presentation effectiveness: data content, expression of decision making, organization, and overall satisfaction. RESULTS: A total of 199 oral case presentations were sampled-112 traditional and 87 AO. Mean length of presentation for traditional presentations was 117 seconds versus 71 seconds for AO presentations (p < 0.001), a clinically significant difference, without significant differences in the essential measures of case presentation effectiveness. CONCLUSIONS: AO oral case presentation may provide a means for emergency medicine residents to "get to the point" and to communicate effectively and efficiently. PMID- 12896885 TI - Information technology in emergency medicine residency-affiliated emergency departments. AB - OBJECTIVES: To describe acquisition and implementation of information technology (IT) in U.S. emergency medicine (EM) residency-affiliated emergency departments (EDs), including automatic medication error checking. METHODS: This was a survey of all U.S. EM residencies active in September 2000. Respondents specified whether specific IT tools had been "acquired" and "implemented fully." EDs were categorized according to primary versus affiliated training site, trauma level, and census. Numbers of "yes" responses were compared according to ED type (Kruskal-Wallis test, p < or = 0.05 significant). RESULTS: Of 121 residency programs, data were obtained from 93 (77%) for a total of 149 EDs. The percentages of EDs that reported full implementation for each technology are as follows: medication error checking, 7%; medication order entry, 18%; nonmedication orders, 7%; clinical documentation, 21%; old electrocardiograms, 62%; laboratory results, 84%; radiography order entry, 62%; image retrieval, 29%; radiologists' interpretations, 67%; cardiology reports, 62%; pathology reports, 70%; surgical reports/dictations, 60%; triage, 34%; tracking, 46%; electronic reference materials, 56%; registration, 84%; accounts, 72%; patient management software package, 20%; voice recognition, 7%. Trauma centers reported more IT tools than nontrauma centers (p = 0.01), and primary training sites reported fewer IT tools than affiliated EDs (p = 0.027). CONCLUSIONS: Incorporation of IT is not uniform in EDs where EM residents train. Acquisition of effective IT tools varies, and implementation lags behind acquisition. Fully implemented IT for medication error checking was reported in 7% of EDs; an additional 12% had acquired IT without implementing it fully. PMID- 12896886 TI - Neighborhood characteristics and emergency department utilization. AB - OBJECTIVES: Neighborhood environments have been linked to the prevalence and incidence rates of a variety of diseases and injuries. This study assessed the relations between neighborhood demographic and socioeconomic characteristics and emergency department (ED) utilization. METHODS: Billing data for the calendar year 2000 for adult patients residing within a one-mile radius of the study ED were geo-coded based on home addresses and merged with neighborhood data from the U.S. Census Bureau. Annual ED visit rates per 100 population were computed for each census block within the study area and analyzed in relation to neighborhood characteristics. RESULTS: The study area consisted of 714 census blocks and 42,278 adult residents, with a total of 16,427 visits to the study ED. Annual ED visit rates increased from 17 visits per 100 population for neighborhoods where less than 50% of the population were African American to 51 visits per 100 population for neighborhoods where all residents were African American (p < 0.001). Annual ED visit rates decreased from 53% for neighborhoods where <20% of housing units were owner-occupied to 27% for neighborhoods with >45% owner occupied housing units (p < 0.001). Multivariate linear regression modeling revealed that proportions of African American residents and owner-occupied housing units were significantly associated with ED utilization, independent of age and gender compositions and the distance between residence and the study ED. CONCLUSIONS: Even within a small geographic area, ED utilization may vary greatly by neighborhood characteristics. Neighborhood racial composition and housing tenure are independent predictors of ED visit rates in this urban community. PMID- 12896887 TI - Emergency department presentations of naltrexone-accelerated detoxification. AB - OBJECTIVES: To analyze emergency department (ED) presentations after naltrexone accelerated detoxification. METHODS: This was a retrospective cohort analysis of patient presentations to Sir Charles Gairdner Hospital Emergency Department over a six-month period (November 2000 to April 2001). RESULTS: During the six-month study period, 42 patients presented to the ED after naltrexone-accelerated detoxification. This represented 7% of patients treated at a single clinic over the same period. Presentation occurred within 24 hours in 40% of cases and within 48 hours in 74%. Clinical features on presentation included gastrointestinal (GI) symptoms (vomiting, 60%; abdominal pain, 55%; diarrhea, 45%), central nervous system [CNS] symptoms (excessive drowsiness, 55%; agitation requiring sedation, 50%), and respiratory symptoms (tachypnea, 33%; respiratory difficulties, 19%). Gastrointestinal symptoms were managed adequately with supportive therapy in most cases (intravenous fluids; antiemetics). Agitation sometimes required large doses of intravenous benzodiazepines (up to 730 mg in 44 hours), one-to-one nursing, and security staff. Two of 14 patients presenting with predominantly CNS disturbance required intubation (14%). Mean in-hospital stay for all patients was 18 hours (range 1 to 92 hours). CONCLUSIONS: A few patients undergoing outpatient naltrexone-accelerated detoxification during a six-month period subsequently required ED management. The clinical features encountered in this group of patients can be subdivided into GI or CNS predominance, with different management strategies. Most presentations can be managed in the ED or an associated observation ward, but departmental resources must be available for one-to-one nursing and security personnel. Patients presenting with agitation should be sedated with benzodiazepines; large doses may be required. Close monitoring of respiratory function is mandatory, and advanced airway management may be required. PMID- 12896888 TI - Prospective study of accuracy and outcome of emergency ultrasound for abdominal aortic aneurysm over two years. AB - Determination of the presence of an abdominal aortic aneurysm (AAA) is essential in the management of the symptomatic emergency department (ED) patient. OBJECTIVES: To identify whether emergency ultrasound of the abdominal aorta (EUS AA) by emergency physicians could accurately determine the presence of AAA and guide ED disposition. METHODS: This was a prospective, observational study at an urban ED with more than 100,000 annual patient visits with consecutive patients enrolled over a two-year period. All patients suspected to have AAA underwent standard ED evaluation consisting of EUS-AA, followed by a confirmatory imaging study or laparotomy. AAA was defined as any measured diameter greater than 3 cm. Demographic data, results of confirmatory testing, and patient outcome were collected by retrospective review. RESULTS: A total of 125 patients had EUS-AA performed over a two-year period. The patient population had the following characteristics: average age 66 years, male 54%, hypertension 56%, coronary artery disease 39%, diabetes 22%, and peripheral vascular disease 14%. Confirmatory tests included radiology ultrasound, 28/125 (22%); abdominal computed tomography, 95/125 (76%); abdominal magnetic resonance imaging, 1/125 (1%); and laparotomy, 1/125 (1%). AAA was diagnosed in 29/125 (23%); of those, 27/29 patients had AAA on confirmatory testing. EUS-AA had 100% sensitivity (95% CI = 89.5 to 100), 98% specificity (95% CI = 92.8 to 99.8), 93% positive predictive value (27/29), and 100% negative predictive value (96/96). Admission rate for the study group overall was 70%. Immediate operative management was considered in 17 of 27 (63%) patients with AAA; ten patients were taken to the operating room. CONCLUSIONS: EUS-AA in a symptomatic population for AAA is sensitive and specific. These data suggest that the presence of AAA on EUS-AA should guide urgent consultation. Emergency physicians were able to exclude AAA regardless of disposition from the ED. PMID- 12896889 TI - Comparative analysis of adult versus adolescent sexual assault: epidemiology and patterns of anogenital injury. AB - OBJECTIVES: To compare the characteristics of sexual assault in pubertal girls (<18 years old) and adults in a community-based population of women presenting to an urban sexual assault clinic. METHODS: This case-series analysis evaluated consecutive female patients presenting to a sexual assault clinic during a three year study period. The clinic is associated with a university-affiliated emergency medicine residency program and is staffed by forensic nurses trained to perform medicolegal examinations using colposcopy with nuclear staining. Patient demographics, assault characteristics, and injury patterns were recorded using a standardized classification system. Data from the two patient groups (adolescents vs. women > or =18 years of age) were analyzed using chi-square test and t-test. RESULTS: A total of 766 cases were identified: 43% of the victims were 13 to 17 years old (mean 15.0 years old), and 57% were older than 17 years old (mean 30.8 years old). Adolescents were more likely to be assaulted by an acquaintance or relative (84% vs. 50%, p < 0.001) and to delay medical evaluation (17 hours vs. 12 hours, p < 0.001) than were older women. Adolescent sexual assault was less likely to involve weapons or physical coercion (29% versus 57%, p < 0.001) and was associated with fewer nongenital injuries (33% vs. 55%, p < 0.001). Adolescents had a greater frequency of anogenital injuries (83% vs. 64%, p < 0.001), however, compared with older women. Common sites of injury in adolescents were posterior, including the fossa navicularis, hymen, fourchette, and labia minora. The injuries showed consistent topologic features, varying with the site and nature of tissue. Adult victims of sexual assault had a less consistent pattern of anogenital injuries with fewer hymenal injuries, greater injury to the perianal area, and widespread erythema. CONCLUSIONS: Of women presenting to an urban sexual assault clinic, 43% were adolescents. The epidemiology of sexual trauma and the pattern of anogenital trauma in this age group are unique and may pose special challenges to emergency health care providers. PMID- 12896890 TI - Clinical pearls: Locked great toe. PMID- 12896891 TI - Inferences from aggregated data. PMID- 12896892 TI - Advanced statistics: applying statistical process control techniques to emergency medicine: a primer for providers. AB - Emergency medicine faces unique challenges in the effort to improve efficiency and effectiveness. Increased patient volumes, decreased emergency department (ED) supply, and an increased emphasis on the ED as a diagnostic center have contributed to poor customer satisfaction and process failures such as diversion/bypass. Statistical process control (SPC) techniques developed in industry offer an empirically based means to understand our work processes and manage by fact. Emphasizing that meaningful quality improvement can occur only when it is exercised by "front-line" providers, this primer presents robust yet accessible SPC concepts and techniques for use in today's ED. PMID- 12896893 TI - Faculty development: how to evaluate the effectiveness of a faculty development program in emergency medicine. AB - Evaluation of the effectiveness of any faculty development program (FDP) is crucial to provide assessment of existing programs and to yield valid recommendations for designing future programs that better address the needs of individual faculty members and the sponsoring institutions. The author chose the validated Kirkpatrick's model with four levels of program outcomes (satisfaction data, learning data, performing data, and career change) to evaluate the effectiveness of an FDP in emergency medicine. Assessment of the effectiveness of this FDP can be performed by sponsoring organizations or by independent evaluators experienced in the field for better objectivity. PMID- 12896894 TI - Assessment for deaths in out-of-hospital heroin overdose patients treated with naloxone who refuse transport. AB - Naloxone frequently is used to treat suspected heroin and opioid overdoses in the out-of-hospital setting. The authors' emergency medical services system has operated a policy of allowing these patients, when successfully treated, to sign out against medical advice (AMA) in the field. OBJECTIVES: To evaluate the safety of this AMA policy. METHODS: This is a retrospective review of out-of-hospital and medical examiner (ME) databases over a five-year period. The authors reviewed all ME cases in which opioid overdoses were listed as contributing to the cause of death. These cases were cross-compared with all patients who received naloxone by field paramedics and then refused transport. The charts were reviewed by dates, times, age, sex, location, and ethnicity when available. RESULTS: There were 998 out-of-hospital patients who received naloxone and refused further treatment and 601 ME cases of opioid overdose deaths. When compared by age, time, date, sex, location, and ethnicity, there were no cases in which a patient was treated by paramedics with naloxone within 12 hours of being found dead of an opioid overdose. CONCLUSIONS: Giving naloxone to patients with heroin overdoses in the field and then allowing them to sign out AMA resulted in no identifiable deaths within this study population. PMID- 12896895 TI - Factors affecting the risk of blood bank specimen hemolysis. AB - OBJECTIVES: To evaluate simultaneously several possible risk factors for blood bank specimen hemolysis. METHODS: This was a prospective cohort study of emergency department and labor and delivery patients to estimate the effect of various factors on the risk of blood bank specimen hemolysis. Study variables included patient demographics, type and gauge of needle or catheter, anatomic location of venipuncture, and patient care area. Hemolysis was determined by blood bank laboratory technicians. Cox proportional hazards multivariate regression modeling was performed to estimate the adjusted relative risks for hemolysis. RESULTS: Of the 605 subjects with complete data, 194 (32.1%) subjects had blood specimens drawn directly with a steel needle, and 411 (69.1%) had specimens drawn through a Vialon (BD Medical Systems, Inc., Sandy, UT) intravenous (IV) angiocatheter. The overall risk of hemolysis for all was 7%, 10% for Vialon IV angiocatheters and 1.5% for steel needles. In the multivariate analysis, the factors most closely associated with hemolysis were the use of Vialon IV catheters and sampling from an anatomic site other than the antecubital area. CONCLUSIONS: Blood bank specimens drawn from Vialon IV catheters (particularly smaller gauge catheters) and from veins outside the antecubital area are at significantly increased risk to hemolyze. PMID- 12896896 TI - U.S. allopathic emergency medicine residency programs: a descriptive review. AB - OBJECTIVE: To use collected data from all 124 allopathic emergency medicine residency (EMR) programs in existence in 2001 to describe allopathic EMR training programs in the United States. METHODS: The authors performed a cross-sectional review of all 124 EMRs using self-reported, standardized data from regularly updated sources: the SAEM Residency Catalog and the AMA's FREIDA (Fellowship and Residency Electronic Interactive Database Access). The authors report median values, ranges, and interquartile ranges (IQRs) on a variety of variables. RESULTS: Median values for EMR programs included a class size of 10 per year; a total faculty-to-resident ratio of 0.78 (1:1.29); an annual primary emergency department (ED) census of 66,000 (IQR = 53,000 to 80,000), with an annual combined primary and secondary ED census of 95,500 (IQR = 75,000 to 126,000); a combined pediatric census of 20,000 (IQR = 15,000 to 32,500) at the primary and secondary sites; an annual combined ED visit per resident of 3,181 (IQR = 2,542 to 3,949), including 733 pediatric visits per resident, at the primary and secondary sites; an admission rate of 20% (IQR = 17% to 23%); and an intensive care unit admission rate of 13% (IQR = 5% to 20%). Despite a wide variability in ranges and IQR in most characteristics pertaining to training setting, the overall reported completion rate of training for residents was high (99%). CONCLUSIONS: This summary description provides median values and ranges for important components of the emergency medicine training environment and provides a basis of comparison for medical students and graduate medical educators. PMID- 12896897 TI - Development and implementation of a relative value scale for teaching in emergency medicine: the teaching value unit. AB - Relative value units exist for measuring clinical productivity. Limited objective measures exist, however, for nonclinical activities, specifically teaching. OBJECTIVE: To develop an objective measure of teaching productivity linked to a performance-based incentive plan. METHODS: Teaching goals and objectives were identified before the 1998-1999 academic year. Teaching value units (TVUs), objective measures for quantifying teaching activities, were developed and assigned based on an estimation of time needed to complete each activity and weighted for importance to the teaching mission. Each physician was allocated teaching time based on past performance and future goals. Targeted TVUs necessary to meet expectations were proportionate to allocated teaching time. Teaching productivity was defined as a percentage of targeted TVUs achieved. Incentive dollars for teaching were distributed based on percentage of targeted TVUs achieved, weighted individually for teaching load. RESULTS: Teaching productivity was evaluated over a three-year period. In year 1, mean TVUs allocated/physician were 181 units (range 25 to 449). Four of 18 physicians (22%) met expectations. The mean individual TVUs achieved were 54% of expected (range 0% to 114%). By year 3, mean TVUs allocated/physician were 179 (range 45 to 629). Twelve of 22 physicians (55%) met expectations. The mean individual TVUs achieved were 82% of expected (range 11% to 146%). Between year 1 and year 3, group productivity increased from 73% to 88%, and mean individual productivity increased from 54% to 82% (p = 0.01). CONCLUSIONS: The development of a TVU-based system enabled objective quantification and monitoring of a broad range of teaching activities. The TVU-based system linked to an incentive plan helped to increase individual and group teaching productivity. PMID- 12896898 TI - The use and safety of droperidol in a high-risk, inner-city emergency department patient population. PMID- 12896899 TI - Medical errors and emergency medicine: will the difficult questions be asked, and answered? PMID- 12896900 TI - Bedside ultrasonography in emergency medicine training programs. PMID- 12896901 TI - Forced expression of antisense 14-3-3beta RNA suppresses tumor cell growth in vitro and in vivo. AB - The 14-3-3 family proteins are key regulators of various signal transduction pathways including malignant transformation. Previously, we found that the expression of the 14-3-3beta gene is deregulated as well as c-myc gene in aflatoxin B1 (AFB1)-induced rat hepatoma K1 and K2 cells. To elucidate the implication of 14-3-3beta in tumor cell growth, in this paper we analyzed the effect of forced expression of antisense 14-3-3beta RNA on the growth and tumorigenicity of K2 cells. K2 cells transfected with antisense 14-3-3beta cDNA expression vector diminished their growth ability in monolayer culture and in semi-solid medium. Expression level of vascular endothelial growth factor mRNA was also reduced in these transfectants. Tumors that formed by the transfectants in nude mice were much smaller and histologically more benign tumors, because of their decreased level of mitosis compared with those of the parental cells. Frequency of apoptosis detected by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling assay was increased in the transfectant-derived tumors accompanying the inhibition of angiogenesis. In addition, over-expression of 14-3 3beta mRNA was observed in various murine tumor cell lines. These results suggest that 14-3-3beta gene plays a pivotal role in abnormal growth of tumor cells in vitro and in vivo. PMID- 12896902 TI - Suppression of gap junctional intercellular communication via 5' CpG island methylation in promoter region of E-cadherin gene in endometrial cancer cells. AB - Previously, we demonstrated that connexins (Cxs) showed aberrant localization and expression in most endometrial hyperplasia and carcinoma samples, indicating that during endometrial carcinogenesis, loss of gap junctional intercellular communication (GJIC) may occur at relatively early stages. In the present study, we focused on the correlations between GJIC and the expression of the E-cadherin and its 5' CpG island methylation in endometrial cancer cells and tissues to investigate their roles in the carcinogenesis and tumor progression of endometrial cancer. In this study, three of the 10 cell lines investigated, Ishikawa, RL-952 and KLE, in which both Cxs and E-cadherin mRNA were expressed, exhibited GJIC by scrape-loading/dye transfer. On the other hand, the other seven cell lines, in which either or both Cxs and E-cadherin mRNA were negative or weakly expressed, did not show GJIC. HEC-50, HEC-1B and HEC-108, in which Cxs were positively expressed but E-cadherin was negatively expressed, showed cytoplasmic localization of Cxs by immunohistochemistry. All five lines, which showed the weak expression of E-cadherin, had E-cadherin 5' CpG island methylation. By immunohistochemistry of 56 endometrial carcinomas, 13 of 27 methylated samples showed weak expression of Cx26 and the other 14 showed diffuse localization in cytoplasm. On the other hand, of 29 unmethylated samples, two showed cell-cell localization, 25 weak expression and two diffuse localization. Furthermore, E-cadherin expression was revealed to be drastically down-regulated by E-cadherin antisense oligonucleotides that post-transcriptionally down regulated E-cadherin expression and in the cell, the localization of Cxs were changed from the cell-cell borders to the cytoplasm, and GJIC also decreased. The results indicated that 5' CpG island methylation, which caused loss of E-cadherin expression, indirectly caused the suppression of GJIC by aberrant localization of Cxs in endometrial carcinoma cells. PMID- 12896903 TI - Expression of glutathione S-transferases (GSTs) in human colon cells and inducibility of GSTM2 by butyrate. AB - The glutathione S-transferases (GSTs) are a multigene family of enzymes largely involved in the detoxification of chemicals. In animals, enhanced expression is mediated by products of gut fermentation. Of these, butyrate induces GSTP1 protein expression and GST activity in the human colon tumor cell line HT29. The aim of the following investigations was to further elucidate butyrate-modulated induction of additional colonic GSTs in HT29 and to determine baseline expression in non-transformed cells, isolated from human colorectal tissue. We measured five GST protein subunits (GSTA1/2-composed of GST A1-1, A1-2 and A2-2-GSTM1, GSTM2, GSTP1, GSTT1) by western blot, GST activity using 1-chloro-2,4-dinitrobenzene as substrate and GSTM2 mRNA expression with RT-PCR. GSTP1, followed by GSTT1, were major subunits in all colon cells. Cells isolated from colon tissue were identified to be colonocytes and colon fibroblasts, both of which also expressed substantial levels of GSTM1 and GSTM2. The inter-individual variation of GST subunits in coloncytes of 15 individuals was marked, with total GST protein per 106 cells differing by more than a factor of four. In HT29, butyrate significantly enhanced GSTA1/2 (3.5-fold), GSTM2 (not detectable in controls), GSTP1 (1.5-fold) and GST activity (1.4-fold), but not GSTM1 or GSTT1. GSTM2 mRNA expression was significantly induced after 24 ( approximately 14-fold) and 72 h treatment ( approximately 8-fold). In colon fibroblasts, butyrate (4 mM, 72 h) also induced GSTM2 protein (1.7-fold) and GST activity (1.4-fold). Colonocytes were too short lived to be used for inducibility studies. In conclusion, GSTs are expressed with high inter-individual variability in human colonocytes. This points to large differences in cellular susceptibility to xenobiotics. However, butyrate, an important luminal component produced from fermentation of dietary fibers, is an efficient inducer of GSTs and especially of GSTM2. This indicates that butyrate may act chemoprotectively by increasing detoxification capabilities in the colon mucosa. PMID- 12896904 TI - Allyl isothiocyanate, a constituent of cruciferous vegetables, inhibits growth of PC-3 human prostate cancer xenografts in vivo. AB - We have shown previously that allyl isothiocyanate (AITC), a constituent of cruciferous vegetables, significantly inhibits survival of PC-3 and LNCaP human prostate cancer cells in culture, whereas proliferation of a normal prostate epithelial cell line is minimally affected by AITC even at concentrations that are highly cytotoxic to the prostate cancer cells. The present studies were designed to test the hypothesis that AITC administration may retard growth of human prostate cancer xenografts in vivo. Bolus i.p. injection of 10 micromol AITC, three times per week (Monday, Wednesday and Friday) beginning the day of tumor cell implantation, significantly inhibited the growth of PC-3 xenograft (P < 0.05 by two-way ANOVA). For example, 26 days after tumor cell implantation, the average tumor volume in control mice (1025 +/- 205 mm3) was approximately 1.7 fold higher compared with AITC-treated mice. Histological analysis of tumors excised at the termination of the experiment revealed a statistically significant increase in number of apoptotic bodies with a concomitant decrease in cells undergoing mitosis in the tumors of AITC-treated mice compared with that of control mice. Western blot analysis indicated an approximately 70% reduction in the levels of anti-apoptotic protein Bcl-2 in the tumor lysate of AITC-treated mice compared with that of control mice. Moreover, the tumors from AITC-treated mice, but not control mice, exhibited cleavage of BID, which is known to promote apoptosis. Statistically significant reduction in the expression of several proteins that regulate G2/M progression, including cyclin B1, cell division cycle (Cdc)25B and Cdc25C (44, 45 and 90% reduction, respectively, compared with control), was also observed in the tumors of AITC-treated mice relative to control tumors. In conclusion, the results of the present study indicate that AITC administration inhibits growth of PC-3 xenografts in vivo by inducing apoptosis and reducing mitotic activity. PMID- 12896905 TI - The alleles of the DNA repair gene O6-alkylguanine-DNA alkyltransferase are expressed at different levels in normal human lung tissue. AB - O6-Alkylguanine-DNA alkyltransferase (MGMT) confers resistance to many of the mutagenic and toxic effects of certain classes of alkylating agents by repairing the DNA lesions responsible. The levels of expression of this protein are of interest in relation to the prevention and treatment of cancer in man. They vary widely between individuals, and the basis of this variation is not understood. RT PCR-RFLP analysis of mRNA from normal human lung tissue reveals that the two MGMT alleles are frequently expressed at different levels, indicating that there is a genetic component to inter-individual variation of MGMT levels and that at least some of this variation maps close to or within the MGMT locus. PMID- 12896906 TI - Cell cycle activation in lung adenocarcinoma cells by the ErbB3/phosphatidylinositol 3-kinase/Akt pathway. AB - Although ErbB3, a member of the epidermal growth factor receptor family, has been implicated in mammary tumorigenesis, investigation of its role in lung tumorigenesis has been limited. We found that ErbB3 was present at high levels in five of seven human lung adenocarcinoma cell lines examined, along with its ligands, heregulins alpha and beta, whereas ErbB3 was absent from HPL1D, a non- transformed cell line from human pulmonary peripheral epithelium. Interactions and effects of ErbB3 were studied in detail in adenocarcinoma lines H441 and H1373. Complexes containing phosphorylated ErbB2, phosphorylated ErbB3 and the p85 regulatory subunit of phosphoinositidyl 3-kinase were detected by co immunoprecipitation experiments and were present constitutively even in the absence of serum-stimulated cell division. Serum treatment increased the pErbB3/p85 complexes and also stimulated phosphorylation of Akt and GSK3beta, increase in cyclin D1 and cell cycle progression, and these events were blocked by the Akt activation inhibitor LY294002. An ErbB3-specific antisense oligonucleotide reduced amounts of ErbB3 protein and p85 complex in both cell lines, and significantly suppressed cell proliferation. These results together suggest involvement of ErbB3 in growth of lung adenocarcinomas, through activation of phosphoinositidyl 3 kinase and Akt, inactivation of GSK3beta and stabilization of cyclin D1 for cell cycle maintenance. It could be a useful therapeutic target. PMID- 12896907 TI - Requirement of a carbon spacer in benzyl isothiocyanate-mediated cytotoxicity and MAPK activation in head and neck squamous cell carcinoma. AB - Cruciferous vegetable-derived isothiocyanates (ITCs; chemical structure: R-N=C=S) are highly effective in affording protection against chemically induced cancers in animal models. Here, we studied the antitumor effects of benzyl isothiocyanate (BITC; Ph-CH2-N=C=S), the predominant ITC compound in broccoli, on head and neck squamous cell carcinoma (HNSCC) cell lines. Proliferation, apoptosis and immunoblotting assays were used to determine the effects and mechanism of several ITCs on HNSCC cells. The IC50 for BITC (24 h treatment) in two of the HNSCC cell lines was approximately 22 and 17 micro M, respectively. Interestingly, phenyl isothiocyanate (PITC; Ph-N=C=S), which is a close structural analog of BITC but lacks a -CH2- spacer that links the aromatic ring to N=C=S moiety, did not result in significant killing of the HNSCC cells in this dose range. BITC (but not PITC) caused activation of caspase 3 and PARP cleavage. Within 20 min of treatment, BITC (but not PITC) induced a rapid activation of p38 MAPK. In addition, BITC (but not PITC) treatment resulted in the activation of p44/42 MAPK. Co-treatment with a specific p38 MAPK inhibitor, SB203580, or an inhibitor of the MEK/MAPK pathway, U0126, partially rescued cells from BITC-induced killing. Our results show that minor structural differences in ITCs can be crucial for the antiproliferative activity of ITCs and that BITC may be a promising chemopreventive as well as therapeutic agent in HNSCC. PMID- 12896908 TI - A case-control study of cyclin D1 CCND1 870A-->G polymorphism and bladder cancer. AB - Expression of cyclin D1 is believed to lead to progression through the G1-S cell cycle checkpoint, and both experimental and pathological evidence suggest that over-expression of this protein may increase the risk of several cancers, including transition cell carcinoma of the bladder. Two major transcripts have been described for CCND1, the gene encoding cyclin D1. CCND1 870A-->G, a common single nucleotide polymorphism in the splice donor region of exon 4, may modulate expression of these transcripts, with the A variant resulting in an increased pool of the isozyme encoded by transcript form b. A statistically significant 1.8 fold increased risk for bladder cancer among individuals possessing the A/A genotype was recently reported in a hospital-based case-control study conducted among native Japanese. We conducted a population-based case-control study of incidence of bladder cancer among non-Hispanic whites in Los Angeles County to examine the relationship between CCND1 870A-->G genotypes and bladder cancer risk. No association between the A/A genotype and risk was observed (odds ratio = 0.90, 95% confidence interval 0.60-1.33). The null association was not appreciably modified by bladder cancer risk factors, including lifetime smoking history, or by histopathologic classification. PMID- 12896909 TI - Identification of genes preferentially expressed in mammary epithelial cells of Copenhagen rat using subtractive hybridization and microarrays. AB - Rats, like humans, vary considerably in susceptibility for mammary cancer development among different strains. The Copenhagen (Cop) rat is extremely resistant to mammary cancer development induced by a variety of carcinogens. Multiple genetic loci have been linked to the resistant phenotype, but the genes have yet to be cloned and the mechanisms underlying the resistance still remain unknown. Transplantation experiments, however, have demonstrated that these genes act only in the epithelial cells of mammary parenchyma; they do not act systemically. In the present study, we analyzed genes differentially expressed in mammary epithelial cells obtained from pubescent female Cop and susceptible Buffalo (Buf) rats, using PCR-based suppressive subtractive hybridization and cDNA microarray approaches. Our results showed a high degree of similarity in the expression profiles of about 4000 genes between Cop and Buf rats, with a few exceptions. We found that the interleukin-2 receptor alpha (IL-2Ralpha) chain gene and claudin-6 gene were preferentially expressed in mammary epithelial cells purified from Cop rats. We further demonstrated that IL-2Ralpha message was undetectable in two rat mammary cancer cell lines and in two human breast cancer cell lines. The level of claudin-6 mRNA was undetectable in two rat mammary cancer cell lines and was lower in two human breast cancer cell lines and one breast cancer sample than that in normal breast tissues. These results suggest that IL-2Ralpha and claudin-6 may function as tumor suppressors, particularly for breast cancer. However, this possibility needs further investigation. PMID- 12896910 TI - Meat and cancer: haemoglobin and haemin in a low-calcium diet promote colorectal carcinogenesis at the aberrant crypt stage in rats. AB - High intake of red meat, but not of white meat, is associated with an increased risk of colon cancer. However, red meat does not promote cancer in rodents. Haemin, added to low-calcium diets, increases colonic proliferation, and haemoglobin, added to high-fat diets, increases the colon tumour incidence in rats, an effect possibly due to peroxyl radicals. We thus speculated that haem might be the promoting agent in meat, and that prevention strategies could use calcium and antioxidants. These hypotheses were tested in rats at the aberrant crypt foci (ACF) stage at 100 days. F344 rats (n = 124) were given an injection of azoxymethane and were then randomized to 11 groups fed with low-calcium (20 micro mol/g) AIN76-based diets, containing 5% safflower oil. Haemin (0.25, 0.5 and 1.5 micro mol/g) or haemoglobin (1.5 and 3 micro mol haem/g) was added to five experimental diets, compared with a control diet without haem. Three other high-haemin diets (1.5 micro mol/g) were supplemented with calcium (250 micro mol/g), antioxidant butylated hydroxyanisole and rutin (0.05% each), and olive oil, which replaced safflower oil. Faecal water was assayed for lipid peroxidation by thiobarbituric acid reactive substances (TBARs) test, and for cytolytic activity. Haemin strikingly increased the ACF size, dose-dependently, from 2.6 to 11.4 crypts/ACF (all P < 0.001). The high-haemin diet also increased the number of ACF per colon (P < 0.001). Promotion was associated with increased faecal water TBARs and cytotoxicity. Calcium, olive oil and antioxidants each inhibited the haemin-induced ACF promotion, and normalized the faecal TBARs and cytotoxicity. The haemoglobin diets increased the number of ACF and faecal TBARs, but not the ACF size or the faecal cytotoxicity. In conclusion, dietary haemin is the most potent known ACF promoter. Haemoglobin is also a potent promoter of colorectal carcinogenesis. The results suggest that myoglobin in red meat could promote colon cancer. Diets high in calcium, or in oxidation-resistant fats, may prevent the possible cancer-promoting effect of red meat. PMID- 12896911 TI - Breast cancer in men. PMID- 12896912 TI - Clinical trials for tomorrow funded by the MRC. PMID- 12896914 TI - The impact of parental psychiatric disorder on children. PMID- 12896915 TI - Tuberculin testing before BCG vaccination. PMID- 12896916 TI - HIV cases rising sharply in India. PMID- 12896917 TI - Liberia faces a humanitarian catastrophe, warn aid agencies. PMID- 12896918 TI - US agrees to cheap drug imports--as Florida officials break fake drugs ring. PMID- 12896920 TI - Company plans to sell snuff as "smokeless tobacco". PMID- 12896921 TI - Presidential commission urges revamp of US mental health system. PMID- 12896923 TI - Asbestos poisoning was covered up by doctors, claims health team. PMID- 12896925 TI - Commons committee calls for more choice over home births. PMID- 12896927 TI - Kidney trafficking is "big business," says Council of Europe. PMID- 12896929 TI - Germany reaches controversial deal on healthcare reform. PMID- 12896933 TI - Fertilisation authority to review "egg giving". PMID- 12896934 TI - Enzyme potentiated desensitisation in treatment of seasonal allergic rhinitis: double blind randomised controlled study. AB - OBJECTIVE: To assess the efficacy of enzyme potentiated desensitisation in the treatment of severe summer hay fever poorly controlled by pharmacotherapy. DESIGN: Double blind randomised placebo controlled parallel group study. SETTING: Hospital in Hampshire. PARTICIPANTS: 183 participants aged between 18 and 64 with a history of severe summer hay fever for at least two years; all were skin prick test positive to timothy grass pollen. 90 randomised to active treatment; 93 randomised to placebo. INTERVENTIONS: Active treatment: two injections of enzyme potentiated desensitisation, given between eight and 11 weeks apart, each comprising 200 Fishman units of beta glucuronidase, 50 pg 1,3-cyclohexanediol, 50 ng protamine sulphate, and a mixed inhaled allergen extract (pollen mixes for trees, grasses, and weeds; allergenic fungal spores; cat and dog danders; dust and storage mites) in a total volume of 0.05 ml of buffered saline. Placebo: two injections of 0.05 ml buffered saline solution. MAIN OUTCOME MEASURES: Proportion of problem-free days; global rhinoconjunctivitis quality of life scores assessed weekly during pollen season. RESULTS: The active treatment group and the placebo group did not differ in the proportion of problem-free days, quality of life scores, symptom severity scores, change in quantitative skin prick provocation threshold, or change in conjunctival provocation threshold. No clinically significant adverse reactions occurred. CONCLUSIONS: Enzyme potentiated desensitisation showed no treatment effect in this study. PMID- 12896935 TI - Impact of supplementing newborn infants with vitamin A on early infant mortality: community based randomised trial in southern India. AB - OBJECTIVE: To assess the impact of supplementing newborn infants with vitamin A on mortality at age 6 months. DESIGN: Community based, randomised, double blind, placebo controlled trial. SETTING: Two rural districts of Tamil Nadu, southern India. PARTICIPANTS: 11 619 newborn infants allocated 24 000 IU oral vitamin A or placebo on days 1 and 2 after delivery. MAIN OUTCOME MEASURE: Primary outcome measure was mortality at age 6 months. RESULTS: Infants in the vitamin A group had a 22% reduction in total mortality (95% confidence interval 4% to 37%) compared with those in the placebo group. Vitamin A had an impact on mortality between two weeks and three months after treatment, with no additional impact after three months. CONCLUSION: Supplementing newborn infants with vitamin A can significantly reduce early infant mortality. PMID- 12896936 TI - Effect of strategies to reduce exposure of infants to environmental tobacco smoke in the home: cross sectional survey. AB - OBJECTIVE: To examine parents' reported knowledge and use of harm reduction strategies to protect their infants from exposure to tobacco smoke in the home, and the relation between reported use of strategies and urinary cotinine to creatinine ratios in the infants. DESIGN: Cross sectional survey. SETTINGS: Coventry and Birmingham. MAIN OUTCOME MEASURES: Parents' reported knowledge and use of harm reduction strategies and urinary cotinine to creatinine ratios in their infants. PARTICIPANTS: 314 smoking households with infants. RESULTS: 86% of parents (264/307) believed that environmental tobacco smoke is harmful, 90% (281/314) believed that infants can be protected from it in the home, and 10% (32/314) were either unaware of measures or reported using none. 65% of parents (205/314) reported using two or more measures, but only 18% (58/314) reported not allowing smoking in the home. No difference was found in mean log e transformed urinary cotinine to creatinine ratio in infants from households that used no measures compared with households that used less strict measures. Mean log cotinine to creatinine ratios were significantly different in households banning smoking in the home compared with those using less strict or no measures. Banning smoking in the home was independently associated with a significant reduction in urinary cotinine to creatinine ratio by a factor of 2.6 (1.6 to 4.2) after adjustment for average household cigarette consumption, tenure, and overcrowding. CONCLUSIONS: Less than a fifth of parents in smoking households ban smoking in the home. Banning smoking was associated with a small but significant reduction in urinary cotinine to creatinine ratio in infants, whereas less strict measures compared with no measures had no effect on the infants' exposure to environmental tobacco smoke. PMID- 12896937 TI - Retrospective review of care and outcomes in young adults with type 1 diabetes. PMID- 12896938 TI - Why surgical patients do not donate tissue for commercial research: review of records. PMID- 12896939 TI - Identifying and managing patients at low risk of bowel cancer in general practice. PMID- 12896940 TI - What happened to the valid POEMs? A survey of review articles on the treatment of type 2 diabetes. AB - OBJECTIVE: To evaluate systematically the review literature on type 2 diabetes to assess transmission of the findings of the United Kingdom prospective diabetes study (UKPDS), an important source of recent valid patient oriented evidence that matters (POEMs). DESIGN: Inception cohort analysis of the recent medical literature. STUDIES REVIEWED: Thirty five reviews on treatment of type 2 diabetes. MAIN OUTCOME MEASURES: Presentation of three types of information from UKPDS in review articles: recommendations based on patient oriented outcomes of study; recommendations contradicted by patient oriented outcomes of study; and recommendations based on disease oriented outcomes for which no patient oriented evidence exists. RESULTS: Only six of the reviews included the POEM that tight blood glucose control had no effect on diabetes related or overall mortality. Just seven mentioned that metformin treatment was associated with decreased mortality. Most (30) of the reviews did not report that diabetic patients with hypertension benefit more from good blood pressure control than good blood glucose control. No review pointed out that treatment of overweight patients with type 2 diabetes with insulin or sulphonylurea drugs had no effect on microvascular or macrovascular outcomes. Thirteen reviews recommended drugs as first line treatment for which we do not have patient oriented outcomes data. The average validity assessment score was 1.3 out of a possible score of 15 (95% confidence interval 0.9 to 1.8). CONCLUSIONS: Review articles on the treatment of type 2 diabetes have not accurately transmitted the valid POEM results of the UKPDS to clinicians. Clinicians relying on review articles written by experts as a source of valid POEMs may be misled. PMID- 12896941 TI - The faults of expert reviews are already well known. PMID- 12896942 TI - Improving compliance with requirements on junior doctors' hours. AB - PROBLEM: Compliance with UK regulations on junior doctors' working hours cannot be achieved by manipulating rotas that maintain existing tiers of cover and work practices. More radical solutions are needed. DESIGN: Audit of change. SETTING: Paediatric night rota in large children's hospital. KEY MEASURES FOR IMPROVEMENT: Compliance with regulations on working hours assessed by diary cards; workload assessed by staff attendance on wards; patient safety assessed through critical incident reports. STRATEGIES FOR CHANGE: Development of new staff roles, followed by change from a partial shift rota comprising 11 doctors and one senior nurse, to a full shift night team comprising three middle grade doctors and two senior nurses. EFFECTS OF CHANGE: Compliance with regulations on working hours increased from 33% to 77%. Workload changed little and was well within the capacity of the new night team. The effect on patient care and on medical staff requires further evaluation. LESSONS LEARNT: Reduction of junior doctors' working hours requires changes to roles, processes, and practices throughout the organisation. PMID- 12896943 TI - New approaches to preventing restenosis. PMID- 12896944 TI - Percutaneous interventional electrophysiology. PMID- 12896945 TI - Modified informed consent procedure: consent to postponed information. PMID- 12896946 TI - Commentary: an imperfect compromise. PMID- 12896947 TI - Co-proxamol and suicide: Co-proxamol should be restricted, not banned. PMID- 12896948 TI - Co-proxamol and suicide: Availability of co-proxamol has been successfully reduced in Doncaster. PMID- 12896949 TI - Co-proxamol and suicide: Licence needs to be changed. PMID- 12896950 TI - Antidepressant prescribing and suicide: Antidepressants do not reduce suicide rates. PMID- 12896951 TI - Antidepressant prescribing and suicide: Analysis is misleading. PMID- 12896952 TI - Antidepressant prescribing and suicide: Decline in suicide rate among older people predates 1991. PMID- 12896953 TI - Antidepressant prescribing and suicide: Associations attribute possible causality inappropriately. PMID- 12896954 TI - Pharmacological heterogeneity limits antidepressant study. PMID- 12896955 TI - European Commission's proposal for a council recommendation on cancer screening. PMID- 12896956 TI - Two groups were different in cardiovascular risk study. PMID- 12896957 TI - Opportunistic screening for Chlamydia: Microbiological input is essential in Chlamydia screening programmes. PMID- 12896958 TI - Guidelines for anecdotes might include more information. PMID- 12896960 TI - Randomise foundation trusts. PMID- 12896961 TI - Time has come for a medical minister of health. PMID- 12896963 TI - Behavioural approaches are helpful in overactive bladder. PMID- 12896964 TI - Working as a GP with a special interest in mental health. PMID- 12896967 TI - Cerebellar deficits and hyperactivity in mice lacking Smad4. AB - Smad4 is a central mediator of TGF-beta signals, which are known to play essential roles in many biological processes. Using a Cre-loxP approach to overcome early embryonic lethality, we have studied functions of TGF-beta/Smad4 signals in the central nervous system (CNS). No obvious deficits were detected in mice carrying the targeted disruption of Smad4 in the CNS. The overall morphology of the hippocampus appeared normal. There was no change in the proliferation of neuronal precursor cells, nor in several forms of synaptic plasticity. In contrast, deletion of Smad4 resulted in a marked decrease in the number of cerebellar Purkinje cells and parvalbumin-positive interneurons. Accompanied by the abnormality in the cerebellum, mutant mice also exhibited significantly increased vertical activity. Thus, our study reveals an unexpected role for Smad4 in cerebellar development and in the control of motor function. PMID- 12896968 TI - Elk-3 is a transcriptional repressor of nitric-oxide synthase 2. AB - The inducible isoform of nitric-oxide synthase (NOS2), a key enzyme catalyzing the dramatic increase in nitric oxide by lipopolysaccharide (LPS), plays an important role in the pathophysiology of endotoxemia and sepsis. Recent evidence suggests that Ets transcription factors may contribute to NOS2 induction by inflammatory stimuli. In this study, we investigated the role of Ets transcription factors in the regulation of NOS2 by LPS and transforming growth factor (TGF)-beta 1. Transient transfection assays in macrophages showed that Ets 2 produced an increase in NOS2 promoter activity, whereas the induction by Ets-1 was modest and NERF2 had no effect. Elk-3 (Net/Erp/Sap-2a) markedly repressed NOS2 promoter activity in a dose-dependent fashion, and overexpression of Elk-3 blunted the induction of endogenous NOS2 message. Mutation of the Net inhibitory domain of Elk-3, but not the C-terminal-binding protein interaction domain, partially alleviated this repressive effect. We also found that deletion of the Ets domain of Elk-3 completely abolished its repressive effect on the NOS2 promoter. LPS administration to macrophages led to a dose-dependent decrease in endogenous Elk-3 mRNA levels, and this decrease in Elk-3 preceded the induction of NOS2 mRNA. In a mouse model of endotoxemia, the expression of Elk-3 in kidney, lung, and heart was significantly down-regulated after systemic administration of LPS, and this down-regulation also preceded NOS2 induction. Moreover, TGF-beta 1 significantly increased endogenous Elk-3 mRNA levels that had been down-regulated by LPS in macrophages. This increase in Elk-3 correlated with a TGF-beta 1 induced down-regulation of NOS2. Taken together, our data suggest that Elk-3 is a strong repressor of NOS2 promoter activity and mRNA levels and that endogenous expression of Elk-3 inversely correlates with NOS2. Thus, Elk-3 may serve as an important mediator of NOS2 gene expression. PMID- 12896969 TI - Mechanism underlying the inner membrane retention of Escherichia coli lipoproteins caused by Lol avoidance signals. AB - Escherichia coli lipoproteins are localized to either the inner or outer membrane depending on the residue at position 2. The inner membrane retention signal, Asp at position 2 in combination with certain residues at position 3, functions as a Lol avoidance signal, i.e. the signal inhibits the recognition of lipoproteins by LolCDE that releases lipoproteins from the inner membrane. To understand the role of the residue at position 2, outer membrane-specific lipoproteins with Cys at position 2 were subjected to chemical modification followed by the release reaction in reconstituted proteoliposomes. Sulfhydryl-specific introduction of nonprotein molecules or a negative charge to Cys did not inhibit the LolCDE dependent release. In contrast, oxidation of Cys to cysteic acid resulted in generation of the Lol avoidance signal, indicating that the Lol avoidance signal requires a critical length of negative charge at the second residue. Furthermore, not only modification of the carboxylic acid of Asp at position 2 but also that of the amine of phosphatidylethanolamine abolished the Lol avoidance function. Based on these results, the Lol avoidance mechanism is discussed. PMID- 12896970 TI - Inactivation of the myocyte enhancer factor-2 repressor histone deacetylase-5 by endogenous Ca(2+) //calmodulin-dependent kinase II promotes depolarization mediated cerebellar granule neuron survival. AB - Cerebellar granule neuron (CGN) survival depends on activity of the myocyte enhancer factor-2 (MEF2) transcription factors. Neuronal MEF2 activity is regulated by depolarization via a mechanism that is presently unclear. Here, we show that depolarization-mediated MEF2 activity and CGN survival are compromised by overexpression of the MEF2 repressor histone deacetylase-5 (HDAC5). Furthermore, removal of depolarization induced rapid cytoplasm-to-nuclear translocation of endogenous HDAC5. This effect was mimicked by addition of the calcium/calmodulin-dependent kinase (CaMK) inhibitor KN93 to depolarizing medium. Removal of depolarization or KN93 addition resulted in dephosphorylation of HDAC5 and its co-precipitation with MEF2D. HDAC5 nuclear translocation triggered by KN93 induced a marked loss of MEF2 activity and subsequent apoptosis. To selectively decrease CaMKII, CGNs were incubated with an antisense oligonucleotide to CaMKIIalpha. This antisense decreased CaMKIIalpha expression and induced nuclear shuttling of HDAC5 in CGNs maintained in depolarizing medium. Selectivity of the CaMKIIalpha antisense was demonstrated by its lack of effect on CaMKIV-mediated CREB phosphorylation. Finally, antisense to CaMKIIalpha induced caspase-3 activation and apoptosis, whereas a missense control oligonucleotide had no effect on CGN survival. These results indicate that depolarization-mediated calcium influx acts through CaMKII to inhibit HDAC5, thereby sustaining high MEF2 activity in CGNs maintained under depolarizing conditions. PMID- 12896971 TI - Targeting and assembly of mitochondrial tail-anchored protein Tom5 to the TOM complex depend on a signal distinct from that of tail-anchored proteins dispersed in the membrane. AB - Mitochondrial outer membrane proteins are synthesized without a cleavable presequence but instead contain segments responsible for mitochondrial targeting and membrane integration within the molecule: the transmembrane segment (TMS) and N- or C-terminal flanking segment. We analyzed targeting and integration of Tom5, a C-tail anchor protein associated with the preprotein translocase of the outer membrane, to the yeast mitochondrial outer membrane in vivo using green fluorescent protein as the reporter and compared the signal with other signals for proteins dispersed in the membrane. The functional assembly of Tom5 into the TOM complex was assessed by blue native PAGE and complementation of temperature sensitive deltatom5 cells. Correct targeting and assembly required (i). an appropriate length TMS rather than hydrophobicity, (ii). a proline residue located at correct position in the TMS and specific residues near the proline, and (iii). that, in contrast to proteins dispersed in the outer membrane, the positive C-terminal segment was dispensable. Based on these findings, we constructed green fluorescent protein fusions with a C-terminal TMS in which the deduced sequences (minimum: Ser-Pro-Met) were inserted at an appropriate position within artificial Leu-Ala repeats. They were targeted to mitochondria and complemented the temperature-sensitive growth phenotype of deltatom5 yeast cells. The membrane-targeting mechanism of Tom5 appears to be distinct from that for proteins that are dispersed in the outer membrane. PMID- 12896972 TI - A p53-independent G1 cell cycle checkpoint induced by the suppression of protein kinase C alpha and theta isoforms. AB - The protein kinase C (PKC) family consists of multiple isoforms that are involved in the regulation of diverse cellular responses. Suppression of PKC induces growth arrest in various types of cells. However, the underlying molecular mechanisms have not been thoroughly investigated. In this report, we demonstrated that the concurrent inhibition, rather than separate inhibition, of phorbol ester dependent PKC alpha and theta isoforms is crucial for the induction of G1 cell cycle arrest and that this negative cell cycle regulation is via p53-independent mechanisms. PKC suppression-mediated growth arrest is associated with the induction of cell cycle inhibitor p21WAF1/CIP1 and the occurrence of hypophosphorylated Rb. The G1 checkpoint induced by the suppression of PKC occurs not only in murine Swiss3T3 but also in p53-deficient cells and human lung cancer cells containing mutated p53. Luciferase and nuclear run-off assays demonstrated that p21WAF1/CIP1 is, in part, transcriptionally regulated in response to the suppression of PKC alpha and theta. However, the stability of p21 mRNA is also augmented after the addition of PKC alpha and theta antisense oligonucleotides, indicating the involvement of post-transcriptional mechanisms in p21WAF1/CIP1 expression. These data suggest the existence of a cell cycle checkpoint pathway regulated by PKC alpha and theta isoforms. Furthermore, our findings support the notion that G1 checkpoint control can be restored in tumor cells containing abnormal p53, by targeting the PKC-regulated p21WAF1/CIP1 induction. PMID- 12896973 TI - Presynaptic N-type calcium channels regulate synaptic growth. AB - Voltage-gated calcium channels couple changes in membrane potential to neuronal functions regulated by calcium, including neurotransmitter release. Here we report that presynaptic N-type calcium channels not only control neurotransmitter release but also regulate synaptic growth at Drosophila neuromuscular junctions. In a screen for behavioral mutants that disrupt synaptic transmission, an allele of the N-type calcium channel locus (Dmca1A) was identified that caused synaptic undergrowth. The underlying molecular defect was identified as a neutralization of a charged residue in the third S4 voltage sensor. RNA interference reduction of N-type calcium channel expression also reduced synaptic growth. Hypomorphic mutations in syntaxin-1A or n-synaptobrevin, which also disrupt neurotransmitter release, did not affect synapse proliferation at the neuromuscular junction, suggesting calcium entry through presynaptic N-type calcium channels, not neurotransmitter release per se, is important for synaptic growth. The reduced synapse proliferation in Dmca1A mutants is not due to increased synapse retraction but instead reflects a role for calcium influx in synaptic growth mechanisms. These results suggest N-type channels participate in synaptic growth through signaling pathways that are distinct from those that mediate neurotransmitter release. Linking presynaptic voltage-gated calcium entry to downstream calcium-sensitive synaptic growth regulators provides an efficient activity-dependent mechanism for modifying synaptic strength. PMID- 12896974 TI - Crystal structure of Escherichia coli PdxA, an enzyme involved in the pyridoxal phosphate biosynthesis pathway. AB - Pyridoxal 5'-phosphate is an essential cofactor for many enzymes responsible for the metabolic conversions of amino acids. Two pathways for its de novo synthesis are known. The pathway utilized by Escherichia coli consists of six enzymatic steps catalyzed by six different enzymes. The fourth step is catalyzed by 4 hydroxythreonine-4-phosphate dehydrogenase (PdxA, E.C. 1.1.1.262), which converts 4-hydroxy-l-threonine phosphate (HTP) to 3-amino-2-oxopropyl phosphate. This divalent metal ion-dependent enzyme has a strict requirement for the phosphate ester form of the substrate HTP, but can utilize either NADP+ or NAD+ as redox cofactor. We report the crystal structure of E. coli PdxA and its complex with HTP and Zn2+. The protein forms tightly bound dimers. Each monomer has an alpha/beta/alpha-fold and can be divided into two subdomains. The active site is located at the dimer interface, within a cleft between the two subdomains and involves residues from both monomers. A Zn2+ ion is bound within each active site, coordinated by three conserved histidine residues from both monomers. In addition two conserved amino acids, Asp247 and Asp267, play a role in maintaining integrity of the active site. The substrate is anchored to the enzyme by the interactions of its phospho group and by coordination of the amino and hydroxyl groups by the Zn2+ ion. PdxA is structurally similar to, but limited in sequence similarity with isocitrate dehydrogenase and isopropylmalate dehydrogenase. These structural similarities and the comparison with a NADP-bound isocitrate dehydrogenase suggest that the cofactor binding mode of PdxA is very similar to that of the other two enzymes and that PdxA catalyzes a stepwise oxidative decarboxylation of the substrate HTP. PMID- 12896975 TI - Merlin links to the cAMP neuronal signaling pathway by anchoring the RIbeta subunit of protein kinase A. AB - The cAMP-protein kinase A (PKA) pathway, important in neuronal signaling, is regulated by molecules that bind and target PKA regulatory subunits. Of four regulatory subunits, RIbeta is most abundantly expressed in brain. The RIbeta knockout mouse has defects in hippocampal synaptic plasticity, suggesting a role for RIbeta in learning and memory-related functions. Molecules that interact with or regulate RIbeta are still unknown. We identified the neurofibromatosis 2 tumor suppressor protein merlin (schwannomin), a molecule related to the ezrin-radixin moesin family of membrane-cytoskeleton linker proteins, as a binding partner for RIbeta. Merlin and RIbeta demonstrated a similar expression pattern in central nervous system neurons and an overlapping subcellular localization in cultured hippocampal neurons and transfected cells. The proteins were coprecipitated from brain lysates by cAMP-agarose and coimmunoprecipited from cellular lysates with specific antibodies. In vitro binding studies verified that the interaction is direct. The interaction appeared to be under conformational regulation and was mediated via the alpha-helical region of merlin. Sequence comparison between merlin and known PKA anchoring proteins identified a conserved alpha-helical PKA anchoring protein motif in merlin. These results identify merlin as the first neuronal binding partner for PKA-RIbeta and suggest a novel function for merlin in connecting neuronal cytoskeleton to PKA signaling. PMID- 12896976 TI - Schizosaccharomyces pombe cells lacking the Ran-binding protein Hba1 show a multidrug resistance phenotype due to constitutive nuclear accumulation of Pap1. AB - In Schizosaccharomyces pombe, the transcription factor Pap1, and the mitogen activated protein kinase Sty1 are excluded from the nucleus in a Crm1-dependent manner under non-stressed conditions. Upon oxidant treatment, both Sty1 and Pap1 concentrate into the nucleus, due to an enhanced import or an impaired export. Hba1, a protein that when overexpressed confers brefeldin A resistance, contains a Ran binding domain. The purpose of this project was to understand at the molecular level the role of Hba1 in the S. pombe oxidative stress response. Fluorescent and confocal microscopy studies demonstrate that Hba1 is located at the nucleoplasm and not at the nuclear envelope. We also demonstrate that either multiple copies or deletion of the hba1 gene induces nuclear accumulation of Pap1 and Sty1. We propose that Hba1 assists Crm1 to export some nuclear export signal containing proteins. Pap1 nuclear accumulation is sufficient for constitutive activation of its specific antioxidant response. On the contrary, constitutive nuclear localization of Sty1 in the Deltahba1 strain does not trigger the Sty1 specific, Atf1-dependent antioxidant response in the absence of stress. We conclude that the increased multidrug resistance of strains lacking or overexpressing Hba1 is due to the accumulation of Pap1 in the nucleus under non stressed conditions. PMID- 12896977 TI - Targeting combinatorial transcriptional complex assembly at specific modules within the interleukin-2 promoter by the immunosuppressant SB203580. AB - The proximal promoter sequence of the interleukin-2 (IL-2) gene contains a series of composite sites or modules that controls much of its responsiveness to environmental stimuli. The integrated targeting of these modules is therefore a major mode of regulation. This report describes how multiple functional hierarchies, required for the recruitment of the p300 co-activator to the CD28RE/AP1 (TRE) module of the IL-2 promoter, are selectively disrupted in human T-cells by the immunosuppressive and anti-inflammatory actions of the p38 mitogen activated protein kinase inhibitor (MAPK), SB203580. The molecular hierarchies targeted by SB203580 include the combinatorial interaction of NF-kappaB and CREB at the CD28RE/AP1 element coupled with the subsequent dynamic co-assembly and activation of p300. Several aspects of this targeting are linked to the ability of SB203580 to inhibit p38 MAPK-controlled pathways. Together, these results provide the molecular basis through which the combinatorial structure and context of the composite elements of the IL-2 promoter dictates mitogen responsiveness and drug susceptibility that are quantitatively and qualitatively distinct from the isolated action of single consensus sequences and/or transcriptional motifs. PMID- 12896978 TI - Characterization of DNA complexes formed by the nuclear receptor constitutive androstane receptor. AB - The nuclear receptor constitutive androstane receptor (CAR) acts as a xenobiotic sensor and regulates the expression of enzymes, such as several cytochromes P450s and the UDP-glucuronosyltransferase (UGT) type 1A1. CAR binds as a heterodimer with the retinoid X receptor (RXR) to specific DNA sites, called response elements (REs). Clusters of CAR REs, referred to as phenobarbital response enhancer modules (PBREMs), have been identified in several CAR target genes. In this study we confirm that REs formed by direct repeats of two AGTTCA hexamers with 4 spacing nucleotides are optimal for the binding of CAR-RXR heterodimers. In addition, we found that the heterodimers also form complexes on everted repeat type arrangements with 8 spacing nucleotides. We also observed that CAR is able to bind DNA as a monomer and to interact in this form with different coregulators even in the presence of RXR. Systematic variation of the nucleotides 5'-flanking to both AGTTCA hexamers showed that the dinucleotide sequence modulates the DNA complex formation of CAR monomers and CAR-RXR heterodimer by a factor of up to 20. The highest preference was found for the sequence AG and lowest for CC. The increased DNA affinity of CAR is mediated by the positively charged arginines 90 and 91 located in the carboxyl-terminal extension of the DNA-binding domain of the receptor. Furthermore, we show that one of the three CAR REs of the human UGT1A1 PBREM is exclusively bound by CAR monomers and this is regulated by ligands that bind to this nuclear receptor. This points to a physiological role for CAR monomers. Therefore, both CAR-RXR heterodimers and CAR monomers can contribute to the gene activating function of PBREMs in CAR target genes. PMID- 12896979 TI - Syntaphilin binds to dynamin-1 and inhibits dynamin-dependent endocytosis. AB - Syntaphilin is a brain-specific syntaxin-binding partner first characterized as an inhibitor of SNARE complex formation and neurotransmitter release. Here we show that syntaphilin also binds to dynamin-1 and through this interaction inhibits dynamin-mediated endocytosis. Immunoprecipitation studies from cross linked rat synaptosomes demonstrate that an endogenous syntaphilin-dynamin-1 complex exists independently of dynamin-1 binding to amphiphysin. Functionally, syntaphilin expression inhibits transferrin internalization in COS-7 cells. These data reveal that syntaphilin is an inhibitor of both SNARE-based fusion and dynamin-mediated endocytosis. PMID- 12896980 TI - Selenite-induced survival of HuH7 hepatoma cells involves activation of focal adhesion kinase-phosphatidylinositol 3-kinase-Akt pathway and Rac1. AB - Selenium has been shown to sustain the growth of selected human hepatocellular carcinoma cell lines under serum-free conditions, but the detailed mechanism remained undetermined. In the present study, the molecular mechanism(s) involving sodium selenite (Na2SO3, Se) as a survival agent were determined. Selenite not only protects HuH7 cells from serum deprivation-induced apoptosis, it also supports its long-term growth in sodium selenite (10(-7)m) supplemented serum free medium. The anti-apoptotic effect correlates with activation of focal adhesion kinase and the phosphatidylinositol 3-kinase (PI3K)-Akt kinase pathway. Using HuH7 cells stably transfected with a constitutively active Akt kinase and PI3K inhibitor LY294002, selenite-induced cell survival was shown to be PI3K-Akt dependent. Parallel changes included a significant reduction in the intracellular reactive oxygen species content, the reversal of DNA fragmentation, and the suppression of caspase and apoptosis signal-regulating kinase 1 activities. HuH7 cells stably expressing a Rac1 mutant N17 (Rac1N17-HuH7) are refractory to selenite treatment. In these cells selenite supplement neither triggers Akt activation nor supports cell proliferation. Participation of Rac1 activation in this event is supported by the fact that selenite treatment drastically enhanced activation of Rac1. The exact link between selenite treatment, Rac1 activation, and activation of the focal adhesion kinase-PI 3-kinase, however, remains to be characterized. The mitogenic signaling mediated by selenite may involve unconventional growth stimuli including higher glutathione peroxidase 1 activity and higher transcription levels of selenoprotein P. The selenium-HuH7 system we have established thus provides a unique tool that will allow the biological role of selenite in growth regulation of hepatocytes to be studied in detail. PMID- 12896981 TI - Opposing roles of C/EBPbeta and AP-1 in the control of fibroblast proliferation and growth arrest-specific gene expression. AB - Chicken embryo fibroblasts (CEF) express several growth arrest-specific (GAS) gene products in G0. In contact-inhibited cells, the expression of the most abundant of these proteins, the p20K lipocalin, is activated at the transcriptional level by C/EBPbeta. In this report, we describe the role of C/EBPbeta in CEF proliferation. We show that the expression of a dominant negative mutant of C/EBPbeta (designated Delta184-C/EBPbeta) completely inhibited p20K expression at confluence and stimulated the proliferation of CEF without inducing transformation. Mouse embryo fibroblasts nullizygous for C/EBPbeta had a proliferative advantage over cells with one or two functional copies of this gene. C/EBP inhibition enhanced the expression of the three major components of AP-1 in cycling CEF, namely c-Jun, JunD, and Fra-2, and stimulated AP-1 activity. In contrast, the over-expression of C/EBPbeta caused a dramatic reduction in the levels of AP-1 proteins. Therefore, C/EBPbeta is a negative regulator of AP-1 expression and activity in CEF. The expression of cyclin D1 and cell proliferation were stimulated by the dominant negative mutant of C/EBPbeta but not in the presence of TAM67, a dominant negative mutant of c-Jun and AP-1. CEF over-expressing c-Jun, and to a lesser extent JunD and Fra-2, did not growth arrest at high cell density and did not express p20K. Therefore, AP-1 interfered with the action of C/EBPbeta at high cell density, indicating that these factors play opposing roles in the control of GAS gene expression and CEF proliferation. PMID- 12896982 TI - A novel nuclear localization signal in the auxiliary domain of apobec-1 complementation factor regulates nucleocytoplasmic import and shuttling. AB - C to U editing of the nuclear apolipoprotein B (apoB) transcript is mediated by a core enzyme containing a catalytic deaminase, apobec-1, and an RNA binding subunit, apobec-1 complementation factor (ACF). ACF expression is predominantly nuclear, including mutant proteins with deletions of a putative nuclear localization signal. We have now identified a novel 41-residue motif (ANS) in the auxiliary domain of ACF that functions as an authentic nuclear localization signal. ANS-green fluorescence protein and ANS-beta-galactosidase chimeras were both expressed exclusively in the nucleus, whereas wild-type chimeras or an ACF deletion mutant lacking the ANS were cytoplasmic. Nuclear accumulation of ACF is transcription-dependent, temperature-sensitive, and reversible, features reminiscent of a shuttling protein. ACF relocates to the cytoplasm after actinomycin D treatment, an effect blocked by the CRM1 inhibitor leptomycin B. Heterokaryon assays confirmed directly that ACF shuttles in vivo. ACF binds to the protein carrier, transportin 2 in vivo, and colocalizes to the nucleus as determined by confocal microscopy. Co-immunoprecipitation experiments revealed that transportin 2 binds directly to the ANS motif. These data suggest that directed nuclear localization and compartmentalization of the core complex of the apoB RNA editing enzyme is regulated through a dominant targeting sequence (ANS) contained within ACF. PMID- 12896983 TI - Oxidative stress reprograms lipopolysaccharide signaling via Src kinase-dependent pathway in RAW 264.7 macrophage cell line. AB - Oxidative stress generated during ischemia/reperfusion injury has been shown to augment cellular responsiveness. Whereas oxidants are themselves known to induce several intracellular signaling cascades, their effect on signaling pathways initiated by other inflammatory stimuli remains poorly elucidated. Previous work has suggested that oxidants are able to prime alveolar macrophages for increased NF-kappa B translocation in response to treatment with lipopolysaccharide (LPS). Because oxidants are known to stimulate the Src family of tyrosine kinases, we hypothesized that the oxidants might contribute to augmented NF-kappa B translocation by LPS via the involvement of Src family kinases. To model macrophage priming in vitro, the murine macrophage cell line, RAW 264.7, was first incubated with various oxidants and then exposed to low dose LPS. These studies show that oxidant stress is able to augment macrophage responsiveness to LPS as evidenced by earlier and increased NF-kappa B translocation. Inhibition of the Src family kinases by either pharmacological inhibition using PP2 or through a molecular approach by cell transfection with Csk was found to prevent the augmented LPS-induced NF-kappa B translocation caused by oxidants. Interestingly, while Src kinase inhibition was able to prevent the LPS-induced NF-kappa B translocation in oxidant-treated macrophages, this strategy had no effect on NF kappa B translocation caused by LPS in the absence of oxidants. These findings suggested that oxidative stress might divert LPS signaling along an alternative signaling pathway. Further studies demonstrated that the Src-dependent pathway induced by oxidant pretreatment involved the activation of phosphatidylinositol 3 kinase. Involvement of this pathway appeared to be independent of traditional LPS signaling. Together, these studies provide a novel potential mechanism whereby oxidants might prime alveolar macrophages for altered responsiveness to subsequent inflammatory stimuli and suggest different cellular targets for immunomodulation following ischemia/reperfusion. PMID- 12896984 TI - Polar localization of CheA2 in Rhodobacter sphaeroides requires specific Che homologs. AB - Rhodobacter sphaeroides is a motile bacterium that has multiple chemotaxis genes organized predominantly in three major operons (cheOp(1), cheOp(2), and cheOp(3)). The chemoreceptor proteins are clustered at two distinct locations, the cell poles and in one or more cytoplasmic clusters. One intriguing possibility is that the physically distinct chemoreceptor clusters are each composed of a defined subset of specific chemotaxis proteins, including the chemoreceptors themselves plus specific CheW and CheA proteins. Here we report the subcellular localization of one such protein, CheA(2), under aerobic and photoheterotrophic growth conditions. CheA(2) is predominantly clustered and localized at the cell poles under both growth conditions. Furthermore, its localization is dependent upon one or more genes in cheOp(2) but not those of cheOp(1) or cheOp(3). In E. coli, the polar localization of CheA depends upon CheW. The R. sphaeroides cheOp(2) contains two cheW genes. Interestingly, CheW(2) is required under both aerobic and photoheterotrophic conditions, whereas CheW(3) is not required under aerobic conditions but appears to play a modest role under photoheterotrophic conditions. This suggests that R. sphaeroides contains at least two distinct chemotaxis complexes, possibly composed of proteins dedicated for each subcellular location. Furthermore, the composition of these spatially distinct complexes may change under different growth conditions. PMID- 12896985 TI - Mutation in the relA gene of Vibrio cholerae affects in vitro and in vivo expression of virulence factors. AB - The relA gene product determines the level of (p)ppGpp, the effector nucleotides of the bacterial stringent response that are also involved in the regulation of other functions, like antibiotic production and quorum sensing. In order to explore the possible involvement of relA in the regulation of virulence of Vibrio cholerae, a relA homolog from the organism (relA(VCH)) was cloned and sequenced. The relA(VCH) gene encodes a 738-amino-acid protein having functions similar to those of other gram-negative bacteria, including Escherichia coli. A deltarelA::kan allele was generated by replacing approximately 31% of the open reading frame of wild-type relA of V. cholerae El Tor strain C6709 with a kanamycin resistance gene. The V. cholerae relA mutant strain thus generated, SHK17, failed to accumulate (p)ppGpp upon amino acid deprivation. Interestingly, compared to the wild type, C6709, the mutant strain SHK17 exhibited significantly reduced in vitro production of two principal virulence factors, cholera toxin (CT) and toxin-coregulated pilus (TCP), under virulence gene-inducing conditions. In vivo experiments carried out in rabbit ileal loop and suckling mouse models also confirmed our in vitro results. The data suggest that (p)ppGpp is essential for maximal expression of CT and TCP during in vitro growth, as well as during intestinal infection by virulent V. cholerae. Northern blot and reverse transcriptase PCR analyses indicated significant reduction in the transcript levels of both virulence factors in the relA mutant strain SHK17. Such marked alteration of virulence phenotypes in SHK17 appears most likely to be due to down regulation of transcript levels of toxR and toxT, the two most important virulence regulatory genes of V. cholerae. In SHK17, the altered expression of the two outer membrane porin proteins, OmpU and OmpT, indicated that the relA mutation most likely affects the ToxR-dependent virulence regulatory pathway, because it had been shown earlier that ToxR directly regulates their expression independently of ToxT. PMID- 12896986 TI - Mutant analysis of the Escherichia coli FhuA protein reveals sites of FhuA activity. AB - The FhuA outer membrane protein of Escherichia coli actively transports ferrichrome, albomycin, and rifamycin CGP 4832, and confers sensitivity to microcin J25, colicin M, and the phages T1, T5, and phi80. Guided by the FhuA crystal structure and derived predictions on how FhuA might function, mutants were isolated in the cork domain (residues 1 to 160) and in the beta-barrel domain (residues 161 to 714). Deletion of the TonB box (residues 7 to 11) completely inactivated all TonB-dependent functions of FhuA. Fixation of the cork to turn 7 of the barrel through a disulfide bridge between introduced C27 and C533 residues abolished ferrichrome transport, which was restored by reduction of the disulfide bond. Deletion of residues 24 to 31, including the switch helix (residues 24 to 29), which upon binding of ferrichrome to FhuA undergoes a large structural transition (17 A) and exposes the N terminus of FhuA (TonB box) to the periplasm, reduced FhuA transport activity (79% of the wild-type activity) but conferred full sensitivity to colicin M and the phages. Duplication of residues 23 to 30 or deletion of residues 13 to 20 resulted in FhuA derivatives with properties similar to those of FhuA with a deletion of residues 24 to 31. However, a frameshift mutation that changed QSEA at positions 18 to 21 to KKAP abolished almost completely most of FhuA's activities. The conserved residues R93 and R133 among energy-coupled outer membrane transporters are thought to fix the cork to the beta-barrel by forming salt bridges to the conserved residues E522 and E571 of the beta-barrel. Proteins with the E522R and E571R mutations were inactive, but inactivity was not caused by repulsion of R93 by R522 and R571 and of R133 by R571. Point mutations in the cork at sites that move or do not move upon the binding of ferrichrome had no effect or conferred only slightly reduced activities. It is concluded that the TonB box is essential for FhuA activity. The TonB box region has to be flexible, but its distance from the cork domain can greatly vary. The removal of salt bridges between the cork and the barrel affects the structure but not the function of FhuA. PMID- 12896987 TI - Detachment of Actinobacillus actinomycetemcomitans biofilm cells by an endogenous beta-hexosaminidase activity. AB - When cultured in broth, fresh clinical isolates of the gram-negative periodontal pathogen Actinobacillus actinomycetemcomitans form tenaciously adherent biofilm colonies on surfaces such as plastic and glass. These biofilm colonies release adherent cells into the medium, and the released cells can attach to the surface of the culture vessel and form new colonies, enabling the biofilm to spread. We mutagenized A. actinomycetemcomitans clinical strain CU1000 with transposon IS903phikan and isolated a transposon insertion mutant that formed biofilm colonies which were tightly adherent to surfaces but which lacked the ability to release cells into the medium and disperse. The transposon insertion in the mutant strain mapped to a gene, designated dspB, that was predicted to encode a secreted protein homologous to the catalytic domain of the family 20 glycosyl hydrolases. A plasmid carrying a wild-type dspB gene restored the ability of biofilm colonies of the mutant strain to disperse. We expressed A. actinomycetemcomitans DspB protein engineered to contain a hexahistidine metal binding site at its C terminus in Escherichia coli and purified the protein by using Ni affinity chromatography. Substrate specificity studies performed with monosaccharides labeled with 4-nitrophenyl groups showed that DspB hydrolyzed the 1-->4 glycosidic bond of beta-substituted N-acetylglucosamine, which is consistent with the known functions of other family 20 glycosyl hydrolases. When added to culture medium, purified DspB protein, but not heat-inactivated DspB, restored the ability of the mutant strain to release cells and disperse. DspB protein also caused the detachment of cells from preformed biofilm colonies of strain CU1000 grown attached to plastic and the disaggregation of highly autoaggregated clumps of CU1000 cells in solution. We concluded that dspB encodes a soluble beta-N-acetylglucosaminidase that causes detachment and dispersion of A. actinomycetemcomitans biofilm cells. PMID- 12896988 TI - Performance of standard phenotypic assays for TonB activity, as evaluated by varying the level of functional, wild-type TonB. AB - The ability of gram-negative bacterial cells to transport cobalamin and iron siderophore complexes and their susceptibility to killing by some bacteriophages and colicins are characteristics routinely used to assay mutations of proteins in the TonB-dependent energy transduction system. These assays vary greatly in sensitivity and are subject to perturbation by overexpression of TonB and, perhaps, other proteins that contribute to the process. Thus, the choice of assay and the means by which a potential mutant is expressed can greatly influence the interpretation and recognition of a given mutant. In the present study, we expressed TonB at several different quantified levels in cells that were then subjected to a panel of assays. Our results suggest that it is reasonable to regard the assays as having windows of sensitivity. Thus, while no single assay satisfactorily spans the potential range of TonB activity, it is evident that certain assays are better suited for resolving small deviations from wild-type levels of activity, with others most useful when activity levels are very low. It is apparent from the results that the application of all possible assays to the characterization of new mutants will yield the most meaningful results. PMID- 12896989 TI - Role of Pseudomonas putida tol-oprL gene products in uptake of solutes through the cytoplasmic membrane. AB - Proteins of the Tol-Pal (Tol-OprL) system play a key role in the maintenance of outer membrane integrity and cell morphology in gram-negative bacteria. Here we describe an additional role for this system in the transport of various carbon sources across the cytoplasmic membrane. Growth of Pseudomonas putida tol-oprL mutant strains in minimal medium with glycerol, fructose, or arginine was impaired, and the growth rate with succinate, proline, or sucrose as the carbon source was lower than the growth rate of the parental strain. Assays with radiolabeled substrates revealed that the rates of uptake of these compounds by mutant cells were lower than the rates of uptake by the wild-type strain. The pattern and amount of outer membrane protein in the P. putida tol-oprL mutants were not changed, suggesting that the transport defect was not in the outer membrane. Consistently, the uptake of radiolabeled glucose and glycerol in spheroplasts was defective in the P. putida tol-oprL mutant strains, suggesting that there was a defect at the cytoplasmic membrane level. Generation of a proton motive force appeared to be unaffected in these mutants. To rule out the possibility that the uptake defect was due to a lack of specific transporter proteins, the PutP symporter was overproduced, but this overproduction did not enhance proline uptake in the tol-oprL mutants. These results suggest that the Tol-OprL system is necessary for appropriate functioning of certain uptake systems at the level of the cytoplasmic membrane. PMID- 12896990 TI - Rod shape determination by the Bacillus subtilis class B penicillin-binding proteins encoded by pbpA and pbpH. AB - The peptidoglycan cell wall determines the shape and structural integrity of a bacterial cell. Class B penicillin-binding proteins (PBPs) carry a transpeptidase activity that cross-links peptidoglycan strands via their peptide side chains, and some of these proteins are directly involved in cell shape determination. No Bacillus subtilis PBP with a clear role in rod shape maintenance has been identified. However, previous studies showed that during outgrowth of pbpA mutant spores, the cells grew in an ovoid shape for several hours before they recovered and took on a normal rod shape. It was postulated that another PBP, expressed later during outgrowth, was able to compensate for the lack of the pbpA product, PBP2a, and to guide the formation of a rod shape. The B. subtilis pbpH (ykuA) gene product is predicted to be a class B PBP with greatest sequence similarity to PBP2a. We found that a pbpH-lacZ fusion was expressed at very low levels in early log phase and increased in late log phase. A pbpH null mutant was indistinguishable from the wild-type, but a pbpA pbpH double mutant was nonviable. When pbpH was placed under the control of an inducible promoter in a pbpA mutant, viability was dependent on pbpH expression. Growth of this strain in the absence of inducer resulted in conversion of the cells from rods to ovoid/round shapes and lysis. We conclude that PBP2a and PbpH play redundant roles in formation of a rod-shaped peptidoglycan cell wall. PMID- 12896991 TI - A rhamnogalacturonan lyase in the Clostridium cellulolyticum cellulosome. AB - Clostridium cellulolyticum secretes large multienzymatic complexes with plant cell wall-degrading activities named cellulosomes. Most of the genes encoding cellulosomal components are located in a large gene cluster: cipC-cel48F-cel8C cel9G-cel9E-orfX-cel9H-cel9J-man5K-cel9M. Downstream of the cel9M gene, a new open reading frame was discovered and named rgl11Y. Amino acid sequence analysis indicates that this gene encodes a multidomain pectinase, Rgl11Y, containing an N terminal signal sequence, a catalytic domain belonging to family 11 of the polysaccharide lyases, and a C-terminal dockerin domain. The present report describes the biochemical characterization of a recombinant form of Rgl11Y. Rgl11Y cleaves the alpha-L-Rhap-(1-->4)-alpha-D-GalpA glycosidic bond in the backbone of rhamnogalacturonan I (RGI) via a beta-elimination mechanism. Its specific activity on potato pectic galactan and rhamnogalacturonan was found to be 28 and 3.6 IU/mg, respectively, indicating that Rgl11Y requires galactan decoration of the RGI backbone. The optimal pH of Rgl11Y is 8.5 and calcium is required for its activity. Rgl11Y was shown to be incorporated in the C. cellulolyticum cellulosome through a typical cohesin-dockerin interaction. Rgl11Y from C. cellulolyticum is the first cellulosomal rhamnogalacturonase characterized. PMID- 12896992 TI - Transcription activation at Escherichia coli FNR-dependent promoters by the gonococcal FNR protein: effects of a novel S18F substitution and comparisons with the corresponding substitution in E. coli FNR. AB - The Neisseria gonorrhoeae genome encodes a homologue of the Escherichia coli FNR protein (the fumarate and nitrate reductase regulator). Despite its similarity to E. coli FNR, the gonococcal FNR only partially complemented an E. coli fnr mutation. After error-prone PCR mutagenesis of the gonococcal fnr gene, we identified four mutant fnr derivatives carrying the same S18F substitution, and we showed that the mutant FNR could activate transcription from a range of class I and class II FNR-dependent promoters in E. coli. Prompted by the similarities between gonococcal and E. coli FNR, we made changes in gonococcal fnr that created substitutions that are equivalent to previously characterized substitutions in E. coli FNR. First, our experiments showed that cysteine, C116, in the gonococcal FNR, equivalent to C122 in E. coli FNR, is essential, presumably because, as in E. coli FNR, it binds to an iron-sulfur center. Second, the L22H and D148A substitutions in gonococcal FNR were made. These changes are equivalent to the L28H and D154A changes in E. coli FNR, which had been shown to increase FNR activity in the presence of oxygen. We show that the effects of these substitutions in gonococcal FNR are distinct from those of the S18F substitution. Similarly, substitutions in the putative activating regions of gonococcal FNR were made. We show that the activity of gonococcal FNR in E. coli can be increased by transplanting certain activating regions from E. coli FNR. The effects of these substitutions are additive to those due to S18F. From these data, we conclude that the effects of the S18F substitution in gonococcal FNR are distinct from the effects of the other substitutions. S18 is immediately adjacent to one of three N-terminal cysteine residues that coordinate the iron-sulfur center, and thus the S18F substitution is most likely to stabilize this center. Support for this came from complementary experiments in which we created the S24F substitution in E. coli FNR, which is equivalent to the S18F substitution in gonococcal FNR. Our results show that the S24F substitution changes the activity of E. coli FNR and that the changes are distinct from those due to previously characterized substitutions. PMID- 12896993 TI - DapE can function as an aspartyl peptidase in the presence of Mn2+. AB - Extracts of a multiply peptidase-deficient (pepNABDPQTE iadA iaaA) Salmonella enterica serovar Typhimurium strain contain an aspartyl dipeptidase activity that is dependent on Mn(2+). Purification of this activity followed by N-terminal sequencing of the protein suggested that the Mn(2+)-dependent peptidase is DapE (N-succinyl-L,L-diaminopimelate desuccinylase). A dapE chromosomal disruption was constructed and transduced into a multiply peptidase-deficient (MPD) strain. Crude extracts of this strain showed no aspartyl peptidase activity, and the strain failed to utilize Asp-Leu as a leucine source. The dapE gene was cloned into expression vectors in order to overproduce either the native protein (DapE) or a hexahistidine fusion protein (DapE-His(6)). Extracts of a strain carrying the plasmid overexpresssing native DapE in the MPD dapE background showed a 3,200 fold elevation of Mn(2+)-dependent aspartyl peptidase activity relative to the MPD dapE(+) strain. In addition, purified DapE-His(6) exhibited Mn(2+)-dependent peptidase activity toward aspartyl dipeptides. Growth of the MPD strain carrying a single genomic copy of dapE on Asp-Leu as a Leu source was slow but detectable. Overproduction of DapE in the MPD dapE strain allowed growth on Asp-Leu at a much faster rate. DapE was found to be specific for N-terminal aspartyl dipeptides: no N-terminal Glu, Met, or Leu peptides were hydrolyzed, nor were any peptides containing more than two amino acids. DapE is known to bind two divalent cations: one with high affinity and the other with lower affinity. Our data indicate that the form of DapE active as a peptidase contains Zn(2+) in the high-affinity site and Mn(2+) in the low-affinity site. PMID- 12896994 TI - In vivo and in vitro evidence that TtgV is the specific regulator of the TtgGHI multidrug and solvent efflux pump of Pseudomonas putida. AB - The TtgGHI efflux pump of Pseudomonas putida DOT-T1E plays a key role in the innate and induced tolerance of this strain to aromatic hydrocarbons and antibiotics. The ttgGHI operon is expressed constitutively from two overlapping promoters in the absence of solvents and at a higher level in their presence, but not in response to antibiotics. Adjacent to the ttgGHI operon is the divergently transcribed ttgVW operon. In TtgV-deficient backgrounds, although not in a TtgW deficient background, expression of the ttgGHI and ttgVW operons increased fourfold. This suggests that TtgV represses expression from the ttgG promoters and controls its own. TtgW plays no major role in the regulation of expression of these promoters. Primer extension revealed that the divergent ttgG and ttgV promoters overlap, and mobility shift assays indicated that TtgV binds to this region with high affinity. DNaseI footprint assays revealed that TtgV protected four DNA helical turns that include the -10 and -35 boxes of the ttgV and ttgG promoters. PMID- 12896995 TI - Transcriptional pausing in the Bacillus subtilis pyr operon in vitro: a role in transcriptional attenuation? AB - The genes encoding the enzymes of pyrimidine nucleotide biosynthesis (pyr genes) are regulated in Bacillus subtilis and many other bacterial species by transcriptional attenuation. When UMP or UTP is bound to the PyrR regulatory protein, it binds to pyr mRNA at specific sequences and secondary structures in the RNA. Binding to this site prevents formation of an antiterminator stem-loop in the RNA and permits formation of a downstream terminator, leading to reduced expression of the pyr genes lying downstream from the terminator. The functioning of several other transcriptional attenuation systems has been shown to involve transcriptional pausing; this observation stimulated us to use single-round transcription of pyr genes to test for formation of paused transcripts in vitro. Using templates with each of the three known B. subtilis pyr attenuation sites, we identified one major pause site in each in which the half-life of the paused transcript was increased four- to sixfold by NusA. In each case pausing at the NusA-stimulated site prevented formation of a complete antiterminator stem-loop, while it resulted in increased time for a PyrR binding loop to form and for PyrR to bind to this loop. Thus, the pausing detected in vitro is potentially capable of playing a role in establishing the correct timing for pyr attenuation in vivo. With two of three pyr templates the combination of NusA with PyrR markedly stimulated termination of transcription at the normal termination sites. This suggests that NusA, by stabilizing pausing, plays a role in termination of pyr transcription in vivo. PMID- 12896996 TI - Expression of the Pseudomonas putida OCT plasmid alkane degradation pathway is modulated by two different global control signals: evidence from continuous cultures. AB - Expression of the genes of the alkane degradation pathway encoded in the Pseudomonas putida OCT plasmid are subject to negative and dominant global control depending on the carbon source used and on the physiological status of the cell. We investigated the signals responsible for this control in chemostat cultures under conditions of nutrient or oxygen limitation. Our results show that this global control is not related to the growth rate and responds to two different signals. One signal is the concentration of the carbon source that generates the repressing effect (true catabolite repression control). The second signal is influenced by the level of expression of the cytochome o ubiquinol oxidase, which in turn depends on factors such as oxygen availability or the carbon source used. Since under carbon limitation conditions the first signal is relieved but the second signal is not, we propose that modulation mediated by the cytochrome o ubiquinol oxidase is not classical catabolite repression control but rather a more general physiological control mechanism. The two signals have an additive, but independent, effect, inhibiting induction of the alkane degradation pathway. PMID- 12896997 TI - PorA represents the major cell wall channel of the Gram-positive bacterium Corynebacterium glutamicum. AB - The cell wall of the gram-positive bacterium Corynebacterium glutamicum contains a channel (porin) for the passage of hydrophilic solutes. The channel-forming polypeptide PorA is a 45-amino-acid acidic polypeptide with an excess of four negatively charged amino acids, which is encoded by the 138-bp gene porA. porA was deleted from the chromosome of C.glutamicum wild-type strain ATCC 13032 to obtain mutant ATCC 13032deltaporA. Southern blot analysis demonstrated that porA was deleted. Lipid bilayer experiments revealed that PorA was not present in the cell wall of the mutant strain. Searches within the known chromosome of C. glutamicum by using National Center for Biotechnology Information BLAST and reverse transcription-PCR showed that no other PorA-like protein is encoded on the chromosome or is expressed in the deletion strain. The porA deletion strain exhibited slower growth and longer growth times than the C. glutamicum wild-type strain. Experiments with different antibiotics revealed that the susceptibility of the mutant strain was much lower than that of the wild-type C. glutamicum strain. The results presented here suggest that PorA represents a major hydrophilic pathway through the cell wall and that C. glutamicum contains cell wall channels which are not related to PorA. PMID- 12896998 TI - Dependence of Helicobacter pylori urease activity on the nickel-sequestering ability of the UreE accessory protein. AB - The Helicobacter pylori ureE gene product was previously shown to be required for urease expression, but its characteristics and role have not been determined. The UreE protein has now been overexpressed in Escherichia coli, purified, and characterized, and three altered versions were expressed to address a nickel sequestering role of UreE. Purified UreE formed a dimer in solution and was capable of binding one nickel ion per dimer. Introduction of an extra copy of ureE into the chromosome of mutants carrying mutations in the Ni maturation proteins HypA and HypB resulted in partial restoration of urease activity (up to 24% of the wild-type levels). Fusion proteins of UreE with increased ability to bind nickel were constructed by adding histidine-rich sequences (His-6 or His-10 to the C terminus and His-10 as a sandwich fusion) to the UreE protein. Each fusion protein was overexpressed in E. coli and purified, and its nickel-binding capacity and affinity were determined. Each construct was also expressed in wild type H. pylori and in hypA and hypB mutant strains for determining in vivo urease activities. The urease activity was increased by introduction of all the engineered versions, with the greatest Ni-sequestering version (the His-6 version) also conferring the greatest urease activity on both the hypA and hypB mutants. The differences in urease activities were not due to differences in the amounts of urease peptides. Addition of His-6 to another expressed protein (triose phosphate isomerase) did not result in stimulation of urease, so urease activation is not related to the level of nonspecific protein-bound nickel. The results indicate a correlation between H. pylori urease activity and the nickel sequestering ability of the UreE accessory protein. PMID- 12896999 TI - In vivo characterization of Escherichia coli ftsZ mutants: effects on Z-ring structure and function. AB - We have characterized the in vivo phenotypes of 17 mutations of Escherichia coli ftsZ. In particular, we determined whether these mutations can complement a null ftsZ phenotype, and we demonstrated that two noncomplementing mutations show partial dominant-negative behavior. We performed immunofluorescence microscopy to determine whether these mutants could assemble into normal or abnormal structures in vivo. The mutants separated into four classes-those that complemented the null and formed normal FtsZ rings, those that complemented the null but formed aberrant FtsZ structures, those that formed aberrant FtsZ structures and did not complement, and those that were unable to form any FtsZ structures. We did not find any mutations that produced nonfunctional Z rings of normal appearance. Surprisingly, some mutants that produced extensively spiraled Z-ring structures divided and grew with a normal doubling time. The analysis was carried out using a complementation system based on an ftsZ deletion strain, a temperature sensitive rescue plasmid, and a complementation vector that placed mutated ftsZ alleles under the control of the pBAD promoter, which offered several advantages over previous systems. PMID- 12897000 TI - Regulation of sialic acid catabolism by the DNA binding protein NanR in Escherichia coli. AB - All Escherichia coli strains so far examined possess a chromosomally encoded nanATEK-yhcH operon for the catabolism of sialic acids. These unique nine-carbon sugars are synthesized primarily by higher eukaryotes and can be used as carbon, nitrogen, and energy sources by a variety of microbial pathogens or commensals. The gene nanR, located immediately upstream of the operon, encodes a protein of the FadR/GntR family that represses nan expression in trans. S1 analysis identified the nan transcriptional start, and DNA footprint analysis showed that NanR binds to a region of approximately 30 bp covering the promoter region. Native (nondenaturing) polyacrylamide gel electrophoresis, mass spectrometry, and chemical cross-linking indicated that NanR forms homodimers in solution. The region protected by NanR contains three tandem repeats of the hexameric sequence GGTATA. Gel shift analysis with purified hexahistidine-tagged or native NanR detected three retarded complexes, suggesting that NanR binds sequentially to the three repeats. Artificial operators carrying different numbers of repeats formed the corresponding number of complexes. Among the sugars tested that were predicted to be products of the nan-encoded system, only the exogenous addition of sialic acid resulted in the dramatic induction of a chromosomal nanA-lacZ fusion or displaced NanR from its operator in vitro. Titration of NanR by the nan promoter region or artificial operators carrying different numbers of the GGTATA repeat on plasmids in this fusion strain supported the binding of the regulator to target DNA in vivo. Together, the results indicate that GGTATA is important for NanR binding, but the precise mechanism remains to be determined. PMID- 12897001 TI - Mannitol-1-phosphate dehydrogenase (MtlD) is required for mannitol and glucitol assimilation in Bacillus subtilis: possible cooperation of mtl and gut operons. AB - We found that mannitol-1-phosphate dehydrogenase (MtlD), a component of the mannitol-specific phosphotransferase system, is required for glucitol assimilation in addition to GutR, GutB, and GutP in Bacillus subtilis. Northern hybridization of total RNA and microarray studies of RNA from cells cultured on glucose, mannitol, and glucitol indicated that mannitol as the sole carbon source induced hyperexpression of the mtl operon, whereas glucitol induced both mtl and gut operons. The B. subtilis mtl operon consists of mtlA (encoding enzyme IICBA(mt1)) and mtlD, and its transcriptional regulator gene, mtlR, is located 14.4 kb downstream from the mtl operon on the chromosome. The mtlA, mtlD, and mtlR mutants disrupted by the introduction of the pMUTin derivatives MTLAd, MTLDd, and MTLRd, respectively, could not grow normally on either mannitol or glucitol. However, the growth of MTLAd on glucitol was enhanced by IPTG (isopropyl-beta-D-thiogalactopyranoside). This mutant has an IPTG-inducible promoter (Pspac promoter) located in mtlA, and this site corresponds to the upstream region of mtlD. Insertion mutants of mtlD harboring the chloramphenicol resistance gene also could not grow on either mannitol or glucitol. In contrast, an insertion mutant of mtlA could grow on glucitol but not on mannitol in the presence or absence of IPTG. MtlR bound to the promoter region of the mtl operon but not to a DNA fragment containing the gut promoter region. PMID- 12897002 TI - The virulence activator AphA links quorum sensing to pathogenesis and physiology in Vibrio cholerae by repressing the expression of a penicillin amidase gene on the small chromosome. AB - Activation of the tcpPH promoter on the Vibrio pathogenicity island by AphA and AphB initiates the Vibrio cholerae virulence cascade and is regulated by quorum sensing through the repressive action of HapR on aphA expression. To further understand how the chromosomally encoded AphA protein activates tcpPH expression, site-directed mutagenesis was used to identify the base pairs critical for AphA binding and transcriptional activation. This analysis revealed a region of partial dyad symmetry, TATGCA-N6-TNCNNA, that is important for both of these activities. Searching the V. cholerae genome for this binding site permitted the identification of a second one upstream of a penicillin V amidase (PVA) gene on the small chromosome. AphA binds to and footprints this site, which overlaps the pva transcriptional start, consistent with its role as a repressor at this promoter. Since aphA expression is under quorum-sensing control, the response regulators LuxO and HapR also influence pva expression. Thus, pva is repressed at low cell density when AphA levels are high, and it is derepressed at high cell density when AphA levels are reduced. Penicillin amidases are thought to function as scavengers for phenylacetylated compounds in the nonparasitic environment. That AphA oppositely regulates the expression of pva from that of virulence, together with the observation that PVA does not play a role in virulence, suggests that these activities are coordinated to serve V. cholerae in different biological niches. PMID- 12897003 TI - Residues C123 and D58 of the 2-methylisocitrate lyase (PrpB) enzyme of Salmonella enterica are essential for catalysis. AB - The prpB gene of Salmonella enterica serovar Typhimurium LT2 encodes a protein with 2-methylisocitrate (2-MIC) lyase activity, which cleaves 2-MIC into pyruvate and succinate during the conversion of propionate to pyruvate via the 2 methylcitric acid cycle. This paper reports the isolation and kinetic characterization of wild-type and five mutant PrpB proteins. Wild-type PrpB protein had a molecular mass of approximately 32 kDa per subunit, and the biologically active enzyme was comprised of four subunits. Optimal 2-MIC lyase activity was measured at pH 7.5 and 50 degrees C, and the reaction required Mg(2+) ions; equimolar concentrations of Mn(2+) ions were a poor substitute for Mg(2+) (28% specific activity). Dithiothreitol (DTT) or reduced glutathione (GSH) was required for optimal activity; the role of DTT or GSH was apparently not to reduce disulfide bonds, since the disulfide-specific reducing agent Tris(2 carboxyethyl)phosphine hydrochloride failed to substitute for DTT or GSH. The K(m) of PrpB for 2-MIC was measured at 19 micro M, with a k(cat) of 105 s(-1). Mutations in the prpB gene were introduced by site-directed mutagenesis based on the active-site residues deemed important for catalysis in the closely related phosphoenolpyruvate mutase and isocitrate lyase enzymes. Residues D58, K121, C123, and H125 of PrpB were changed to alanine, and residue R122 was changed to lysine. Nondenaturing polyacrylamide gel electrophoresis indicated that all mutant PrpB proteins retained the same oligomeric state of the wild-type enzyme, which is known to form tetramers. The PrpB(K121A), PrpB(H125A), and PrpB(R122K) mutant proteins formed enzymes that had 1,050-, 750-, and 2-fold decreases in k(cat) for 2-MIC lyase activity, respectively. The PrpB(D58A) and PrpB(C123A) proteins formed tetramers that displayed no detectable 2-MIC lyase activity indicating that both of these residues are essential for catalysis. Based on the proposed mechanism of the closely related isocitrate lyases, PrpB residue C123 is proposed to serve as the active site base, and residue D58 is critical for the coordination of a required Mg(2+) ion. PMID- 12897004 TI - Allosteric regulation of Bacillus subtilis NAD kinase by quinolinic acid. AB - NADP is essential for biosynthetic pathways, energy, and signal transduction. In living organisms, NADP biosynthesis proceeds through the phosphorylation of NAD with a reaction catalyzed by NAD kinase. We expressed, purified, and characterized Bacillus subtilis NAD kinase. This enzyme represents a new member of the inorganic polyphosphate [poly(P)]/ATP NAD kinase subfamily, as it can use poly(P), ATP, or other nucleoside triphosphates as phosphoryl donors. NAD kinase showed marked positive cooperativity for the substrates ATP and poly(P) and was inhibited by its product, NADP, suggesting that the enzyme plays a major regulatory role in NADP biosynthesis. We discovered that quinolinic acid, a central metabolite in NAD(P) biosynthesis, behaved like a strong allosteric activator for the enzyme. Therefore, we propose that NAD kinase is a key enzyme for both NADP metabolism and quinolinic acid metabolism. PMID- 12897005 TI - Autoinducer 2 production by Streptococcus gordonii DL1 and the biofilm phenotype of a luxS mutant are influenced by nutritional conditions. AB - The luxS gene, present in many bacterial genera, encodes the autoinducer 2 (AI-2) synthase. AI-2 has been implicated in bacterial signaling, and this study investigated its role in biofilm formation by Streptococcus gordonii, an organism that colonizes human tooth enamel within the first few hours after professional cleaning. Northern blotting and primer extension analyses revealed that S. gordonii luxS is monocistronic. AI-2 production was dependent on nutritional conditions, and maximum AI-2 induction was detected when S. gordonii was grown in the presence of serum and carbonate. In planktonic cultures, AI-2 production rose sharply during the transition from exponential to stationary phase, and the AI-2 concentration peaked approximately 4 h into stationary phase. An S. gordonii luxS mutant that did not produce AI-2 was constructed by homologous recombination. Complementation of the mutant by insertion of an intact luxS gene into the chromosome in tandem with the disrupted gene restored AI-2 production to a level similar to that of the wild-type strain. In planktonic culture, no growth differences were observed between the mutant and wild-type strains when five different media were used. However, when grown for 4 h as biofilms in 25% human saliva under flow, the luxS mutant formed tall microcolonies that differed from those formed by the wild-type and complemented mutant strains. Biofilms of the luxS mutant exhibited finger-like projections of cells that extended into the flow cell lumen. Thus, the inability to produce AI-2 is associated with altered microcolony architecture within S. gordonii biofilms formed in saliva during a time frame consistent with initial colonization of freshly cleaned enamel surfaces. PMID- 12897006 TI - Molecular analysis of Phr peptide processing in Bacillus subtilis. AB - In Bacillus subtilis, an export-import pathway regulates production of the Phr pentapeptide inhibitors of Rap proteins. Processing of the Phr precursor proteins into the active pentapeptide form is a key event in the initiation of sporulation and competence development. The PhrA (ARNQT) and PhrE (SRNVT) peptides inhibit the RapA and RapE phosphatases, respectively, whose activity is directed toward the Spo0F approximately P intermediate response regulator of the sporulation phosphorelay. The PhrC (ERGMT) peptide inhibits the RapC protein acting on the ComA response regulator for competence with regard to DNA transformation. The structural organization of PhrA, PhrE, and PhrC suggested a role for type I signal peptidases in the processing of the Phr preinhibitor, encoded by the phr genes, into the proinhibitor form. The proinhibitor was then postulated to be cleaved to the active pentapeptide inhibitor by an additional enzyme. In this report, we provide evidence that Phr preinhibitor proteins are subject to only one processing event at the peptide bond on the amino-terminal end of the pentapeptide. This processing event is most likely independent of type I signal peptidase activity. In vivo and in vitro analyses indicate that none of the five signal peptidases of B. subtilis (SipS, SipT, SipU, SipV, and SipW) are indispensable for Phr processing. However, we show that SipV and SipT have a previously undescribed role in sporulation, competence, and cell growth. PMID- 12897007 TI - Probing conservation of HAMP linker structure and signal transduction mechanism through analysis of hybrid sensor kinases. AB - The HAMP linker, a predicted structural element observed in many sensor kinases and methyl-accepting chemotaxis proteins, transmits signals between sensory input modules and output modules. HAMP linkers are located immediately inside the cytoplasmic membrane and are predicted to form two short amphipathic alpha helices (AS-1 and AS-2) joined by an unstructured connector. HAMP linkers are found in the Escherichia coli nitrate- and nitrite-responsive sensor kinases NarX and NarQ (which respond to ligand by increasing kinase activity) and the sensor kinase CpxA (which responds to ligand by decreasing kinase activity). We constructed a series of hybrids with fusion points throughout the HAMP linker, in which the sensory modules of NarX or NarQ are fused to the transmitter modules of NarX, NarQ, or CpxA. A hybrid of the NarX sensor module and the CpxA HAMP linker and transmitter module (NarX-CpxA-1) responded to nitrate by decreasing kinase activity, whereas a hybrid in which the HAMP linker of NarX was replaced by that of CpxA (NarX-CpxA-NarX-1) responded to nitrate by increasing kinase activity. However, sequence variations between HAMP linkers do not allow free exchange of HAMP linkers or their components. Certain deletions in the NarX HAMP linker resulted in characteristic abnormal responses to ligand; similar deletions in the NarQ and NarX-CpxA-1 HAMP linkers resulted in responses to ligand generally similar to those seen in NarX. We conclude that the structure and action of the HAMP linker are conserved and that the HAMP linker transmits a signal to the output domain that ligand is bound. PMID- 12897008 TI - Regulation of the Bacillus subtilis extracytoplasmic function protein sigma(Y) and its target promoters. AB - The Bacillus subtilis extracytoplasmic function sigma factor sigma(Y) is of unknown function. We demonstrate that the sigY operon is expressed from an autoregulatory promoter site, P(Y). We selected for transposon-induced mutations that upregulate P(Y) transcription in an attempt to identify genes involved in sigma(Y) regulation. The resulting insertions disrupted yxlC, the gene immediately downstream of sigY. However, the phenotype of the yxlC::Tn10 insertion was due to polarity on the downstream genes of the sigY operon; a nonpolar insertion in yxlC did not lead to derepression of P(Y). Further analyses revealed that both yxlD and yxlE encoded proteins important for the negative regulation of sigma(Y) activity. A comparison of the transcriptomes of wild-type and yxlC::Tn10 mutant strains revealed elevated expression of several operons. However, only one additional gene, ybgB, was unambiguously identified as a direct target for sigma(Y). This was supported by analysis of direct targets for sigma(Y) transcription with whole-genome runoff transcription followed by macroarray analysis. PMID- 12897009 TI - A novel IS element, IS621, of the IS110/IS492 family transposes to a specific site in repetitive extragenic palindromic sequences in Escherichia coli. AB - An Escherichia coli strain, ECOR28, was found to have insertions of an identical sequence (1,279 bp in length) at 10 loci in its genome. This insertion sequence (named IS621) has one large open reading frame encoding a putative protein that is 326 amino acids in length. A computer-aided homology search using the DNA sequence as the query revealed that IS621 was homologous to the piv genes, encoding pilin gene invertase (PIV). A homology search using the amino acid sequence of the putative protein encoded by IS621 as the query revealed that the protein also has partial homology to transposases encoded by the IS110/IS492 family elements, which were known to have partial homology to PIV. This indicates that IS621 belongs to the IS110/IS492 family but is most closely related to the piv genes. In fact, a phylogenetic tree constructed on the basis of amino acid sequences of PIV proteins and transposases revealed that IS621 belongs to the piv gene group, which is distinct from the IS110/IS492 family elements, which form several groups. PIV proteins and transposases encoded by the IS110/IS492 family elements, including IS621, have four acidic amino acid residues, which are conserved at positions in their N-terminal regions. These residues may constitute a tetrad D-E(or D)-D-D motif as the catalytic center. Interestingly, IS621 was inserted at specific sites within repetitive extragenic palindromic (REP) sequences at 10 loci in the ECOR28 genome. IS621 may not recognize the entire REP sequence in transposition, but it recognizes a 15-bp sequence conserved in the REP sequences around the target site. There are several elements belonging to the IS110/IS492 family that also transpose to specific sites in the repeated sequences, as does IS621. IS621 does not have terminal inverted repeats like most of the IS110/IS492 family elements. The terminal sequences of IS621 have homology with the 26-bp inverted repeat sequences of pilin gene inversion sites that are recognized and used for inversion of pilin genes by PIV. This suggests that IS621 initiates transposition through recognition of their terminal regions and cleavage at the ends by a mechanism similar to that used for PIV to promote inversion at the pilin gene inversion sites. PMID- 12897010 TI - Temperature-dependent hypermutational phenotype in recA mutants of Thermus thermophilus HB27. AB - The recA gene from Thermus thermophilus HB27 was cloned and engineered to obtain insertion (recA::kat) and deletion (deltarecA) derivatives. Transcription of recA in this extreme thermophile was induced by mitomycin C, leading to the synthesis of a monocistronic mRNA. This DNA damage-mediated induction was dependent on the integrity of recA. In addition to UV sensitivity, the recA mutants of T. thermophilus showed severe pleiotropic defects, ranging from irregular nucleoid condensation and segregation to a dramatic reduction in viability during culture. An increase in the frequency of both carotenoidless and auxotrophic mutants within surviving cells of the deltarecA strain indicated a high mutation rate. As RecA is not required for plasmid transformation, we have used the alpha-lacZ gene fragment and the ampicillin resistance gene from Escherichia coli as passenger reporters to confirm such high mutation rates. Our data support the idea that the absence of RecA results in a hypermutational phenotype in T. thermophilus. Furthermore, a direct relationship is deduced between the growth temperature and mutation rate, which finally has a deleterious effect on cell survival in the absence of RecA. PMID- 12897011 TI - Identification of CpxR as a positive regulator of icm and dot virulence genes of Legionella pneumophila. AB - To date, 24 Legionella pneumophila genes (icm and dot genes) have been shown to be required for intercellular growth and host cell killing. A previous report indicated that the regulation of these genes is complicated and probably involves several regulatory proteins. In this study, a genetic screen performed in Escherichia coli identified the CpxR response regulator as an activator of the L. pneumophila icmR gene. Construction of an L. pneumophila cpxR insertion mutant showed that the expression of icmR is regulated by CpxR. In addition, a conserved CpxR binding site (GTAAA) was identified in the icmR regulatory region and L. pneumophila His-tagged CpxR protein was shown to bind to the icmR regulatory region using a mobility shift assay. Besides its dramatic effect on the icmR level of expression, the CpxR regulator was also found to affect the expression of the icmV-dotA and icmW-icmX operons, but to a lesser extent. The role of CpxA, the cognate sensor kinase of CpxR, was also examined and its effect on the icmR level of expression was found to be less pronounced than the effect of CpxR. The RpoE sigma factor, which was shown to coregulate genes together with CpxR, was examined as well, but it did not influence icm and dot gene expression. In addition, when the cpxR mutant strain, in which the expression of the icmR gene was dramatically reduced, and the cpxA and rpoE mutant strains were examined for their ability to grow inside Acanthamoeba castellanii and HL-60-derived human macrophages, no intracellular growth defect was observed. This study presents the first evidence for a direct regulator (CpxR) of an icm-dot virulence gene (icmR). The CpxR regulator together with other regulatory factors probably concerts with the expression of icm and dot genes to result in successful infection. PMID- 12897012 TI - Benzoate-coenzyme A ligase from Thauera aromatica: an enzyme acting in anaerobic and aerobic pathways. AB - In the denitrifying member of the beta-Proteobacteria Thauera aromatica, the anaerobic metabolism of aromatic acids such as benzoate or 2-aminobenzoate is initiated by the formation of the coenzyme A (CoA) thioester, benzoyl-CoA and 2 aminobenzoyl-CoA, respectively. Both aromatic substrates were transformed to the acyl-CoA intermediate by a single CoA ligase (AMP forming) that preferentially acted on benzoate. This benzoate-CoA ligase was purified and characterized as a 57-kDa monomeric protein. Based on V(max)/K(m), the specificity constant for 2 aminobenzoate was 15 times lower than that for benzoate; this may be the reason for the slower growth on 2-aminobenzoate. The benzoate-CoA ligase gene was cloned and sequenced and was found not to be part of the gene cluster encoding the general benzoyl-CoA pathway of anaerobic aromatic metabolism. Rather, it was located in a cluster of genes coding for a novel aerobic benzoate oxidation pathway. In line with this finding, the same CoA ligase was induced during aerobic growth with benzoate. A deletion mutant not only was unable to grow anaerobically on benzoate or 2-aminobenzoate, but also aerobic growth on benzoate was affected. This suggests that benzoate induces a single benzoate-CoA ligase. The product of benzoate activation, benzoyl-CoA, then acts as inducer of separate anaerobic or aerobic pathways of benzoyl-CoA, depending on whether oxygen is lacking or present. PMID- 12897013 TI - Haemophilus influenzae Rd lacks a stringently conserved fatty acid biosynthetic enzyme and thermal control of membrane lipid composition. AB - The organization of the fatty acid synthetic genes of Haemophilus influenzae Rd is remarkably similar to that of the paradigm organism, Escherichia coli K-12, except that no homologue of the E. coli fabF gene is present. This finding is unexpected, since fabF is very widely distributed among bacteria and is thought to be the generic 3-ketoacyl-acyl carrier protein (ACP) synthase active on long chain-length substrates. However, H. influenzae Rd contains a homologue of the E. coli fabB gene, which encodes a 3-ketoacyl-ACP synthase required for unsaturated fatty acid synthesis, and it seemed possible that the H. influenzae FabB homologue might have acquired the functions of FabF. E. coli mutants lacking fabF function are unable to regulate the compositions of membrane phospholipids in response to growth temperature. We report in vivo evidence that the enzyme encoded by the H. influenzae fabB gene has properties essentially identical to those of E. coli FabB and lacks FabF activity. Therefore, H. influenzae grows without FabF function. Moreover, as predicted from studies of the E. coli fabF mutants, H. influenzae is unable to change the fatty acid compositions of its membrane phospholipids with growth temperature. We also demonstrate that the fabB gene of Vibrio cholerae El Tor N16961 does not contain a frameshift mutation as was previously reported. PMID- 12897014 TI - Conformation of a bactericidal domain of puroindoline a: structure and mechanism of action of a 13-residue antimicrobial peptide. AB - Puroindoline a, a wheat endosperm-specific protein containing a tryptophan-rich domain, was reported to have antimicrobial activities. We found that a 13-residue fragment of puroindoline a (FPVTWRWWKWWKG-NH(2)) (puroA) exhibits activity against both gram-positive and gram-negative bacteria. This suggests that puroA may be a bactericidal domain of puroindoline a. PuroA interacted strongly with negatively charged phospholipid vesicles and induced efficient dye release from these vesicles, suggesting that the microbicidal effect of puroA may be due to interactions with bacterial membranes. A variety of biophysical and biochemical methods, including fluorescence spectroscopy and microcalorimetry, were used to examine the mode of action of puroA. These studies showed that puroA is located at the membrane interface, probably due to its high content of Trp residues that have a high propensity to partition into the membrane interface. The penetration of these Trp residues in negatively charged phospholipid vesicles resembling bacterial membranes was more extensive than the penetration in neutral vesicles mimicking eukaryotic membranes. Peptide binding had a significant influence on the phase behavior of the former vesicles. The three-dimensional structure of micelle-bound puroA determined by two-dimensional nuclear magnetic resonance spectroscopy indicated that all the positively charged residues are oriented close to the face of Trp indole rings, forming energetically favorable cation-pi interactions. This characteristic, along with its well-defined amphipathic structure upon binding to membrane mimetic systems, allows puroA to insert more deeply into bacterial membranes and disrupt the regular membrane bilayer structure. PMID- 12897015 TI - Mapping the MinE site involved in interaction with the MinD division site selection protein of Escherichia coli. AB - Interactions between the MinD and MinE proteins are required for proper placement of the Escherichia coli division septum. The site within MinE that is required for interaction with MinD was mapped by studying the effects of site-directed minE mutations on MinD-MinE interactions in yeast two-hybrid and three-hybrid experiments. This confirmed that the MinE N-terminal domain is responsible for the interaction of MinE with MinD. Mutations that interfered with the interaction defined an extended surface on one face of the alpha-helical region of the MinE N terminal domain, consistent with the idea that the MinE-MinD interaction involves formation of a coiled-coil structure by interaction with a complementary helical surface within MinD. PMID- 12897016 TI - Phenotype microarray analysis of Escherichia coli K-12 mutants with deletions of all two-component systems. AB - Two-component systems are the most common mechanism of transmembrane signal transduction in bacteria. A typical system consists of a histidine kinase and a partner response regulator. The histidine kinase senses an environmental signal, which it transmits to its partner response regulator via a series of autophosphorylation, phosphotransfer, and dephosphorylation reactions. Much work has been done on particular systems, including several systems with regulatory roles in cellular physiology, communication, development, and, in the case of bacterial pathogens, the expression of genes important for virulence. We used two methods to investigate two-component regulatory systems in Escherichia coli K-12. First, we systematically constructed mutants with deletions of all two-component systems by using a now-standard technique of gene disruption (K. A. Datsenko and B. L. Wanner, Proc. Natl. Acad. Sci. USA 97:6640-6645, 2000). We then analyzed these deletion mutants with a new technology called Phenotype MicroArrays, which permits assays of nearly 2,000 growth phenotypes simultaneously. In this study we tested 100 mutants, including mutants with individual deletions of all two component systems and several related genes, including creBC-regulated genes (cbrA and cbrBC), phoBR-regulated genes (phoA, phoH, phnCDEFGHIJKLMNOP, psiE, and ugpBAECQ), csgD, luxS, and rpoS. The results of this battery of nearly 200,000 tests provided a wealth of new information concerning many of these systems. Of 37 different two-component mutants, 22 showed altered phenotypes. Many phenotypes were expected, and several new phenotypes were also revealed. The results are discussed in terms of the biological roles and other information concerning these systems, including DNA microarray data for a large number of the same mutants. Other mutational effects are also discussed. PMID- 12897017 TI - Genomic profiling of iron-responsive genes in Salmonella enterica serovar typhimurium by high-throughput screening of a random promoter library. AB - The importance of iron to bacteria is shown by the presence of numerous iron scavenging and transport systems and by many genes whose expression is tightly regulated by iron availability. We have taken a global approach to gene expression analysis of Salmonella enterica serovar Typhimurium in response to iron by combining efficient, high-throughput methods with sensitive, luminescent reporting of gene expression using a random promoter library. Real-time expression profiles of the library were generated under low- and high-iron conditions to identify iron-regulated promoters, including a number of previously identified genes. Our results indicate that approximately 7% of the genome may be regulated directly or indirectly by iron. Further analysis of these clones using a Fur titration assay revealed three separate classes of genes; two of these classes consist of Fur-regulated genes. A third class was Fur independent and included both negatively and positively iron-responsive genes. These may reflect new iron-dependent regulons. Iron-responsive genes included iron transporters, iron storage and mobility proteins, iron-containing proteins (redox proteins, oxidoreductases, and cytochromes), transcriptional regulators, and the energy transducer tonB. By identifying a wide variety of iron-responsive genes, we extend our understanding of the global effect of iron availability on gene expression in the bacterial cell. PMID- 12897018 TI - Identification of a regulated alkaline phosphatase, a cell surface-associated lipoprotein, in Mycobacterium smegmatis. AB - Although alkaline phosphatases are common in a wide variety of bacteria, there has been no prior evidence for alkaline phosphatases in Mycobacterium smegmatis. Here we report that transposon insertions in the pst operon, encoding homologues of an inorganic phosphate transporter, leads to constitutive expression of a protein with alkaline phosphatase activity. DNA sequence analysis revealed that M. smegmatis does indeed have a phoA gene that shows high homology to other phoA genes. The M. smegmatis phoA gene was shown to be induced by phosphate starvation and thus negatively regulated by the pst operon. Interestingly, the putative M. smegmatis PhoA has a hydrophobic N-terminal domain which resembles a lipoprotein signal sequence. The M. smegmatis PhoA was demonstrated to be an exported protein associated with the cell surface. Furthermore, immunoprecipitation of PhoA from [(14)C]acetate-labeled M. smegmatis cell lysates demonstrated that this phosphatase is a lipoprotein. PMID- 12897019 TI - A Burkholderia pseudomallei type III secreted protein, BopE, facilitates bacterial invasion of epithelial cells and exhibits guanine nucleotide exchange factor activity. AB - We report the characterization of BopE, a type III secreted protein that is encoded adjacent to the Burkholderia pseudomallei bsa locus and is homologous to Salmonella enterica SopE/SopE2. Inactivation of bopE impaired bacterial entry into HeLa cells, indicating that BopE facilitates invasion. Consistent with this notion, BopE expressed in eukaryotic cells induced rearrangements in the subcortical actin cytoskeleton, and purified BopE exhibited guanine nucleotide exchange factor activity for Cdc42 and Rac1 in vitro. PMID- 12897020 TI - Identification of long intergenic repeat sequences associated with DNA methylation sites in Caulobacter crescentus and other alpha-proteobacteria. AB - A systematic search for motifs associated with CcrM DNA methylation sites revealed four long (>100-bp) motifs (CIR sequences) present in up to 21 copies in Caulobacter crescentus. The CIR1 and CIR2 motifs exhibit a conserved inverted repeat organization, with a CcrM site in the center of one of the repeats. PMID- 12897021 TI - Mutational analysis of cell wall biosynthesis in Mycobacterium avium. AB - The cell wall of the environmental pathogen Mycobacterium avium is important to its virulence and intrinsic antimicrobial resistance. To identify genes involved in cell wall biosynthesis, "transposome" insertion libraries were screened for mutants with altered colony morphology on medium containing the lipoprotein stain Congo red. Nineteen such mutants were isolated and mapped, including 10 with insertions in a functional island of cell wall biosynthetic genes that spans approximately 40 kb of the M. avium genome. PMID- 12897022 TI - The Product of the fimI gene is necessary for Escherichia coli type 1 pilus biosynthesis. AB - Site-directed mutagenesis was employed to create lesions in fimI, a gene of uncertain function located in the chromosomal gene cluster (fim) involved in Escherichia coli type 1 pilus biosynthesis. Chromosomal fimI mutations produced a piliation-negative phenotype. Complementation analysis indicated that a fimI'-kan insertion mutation and a fimI frameshift mutation produced polarity-like effects not seen with an in-frame fimI deletion mutation. Minicell analysis associated fimI with a 16.4-kDa noncytoplasmic protein product (FimI). We conclude that FimI has a required role in normal pilus biosynthesis. PMID- 12897023 TI - Regulated expression of the Escherichia coli dam gene. AB - Regulated expression of the Escherichia coli dam gene has been achieved with the araBAD promoter lacking a ribosome binding site. Cultures of dam mutants containing plasmid pMQ430 show no detectable methylation in the absence of arabinose and complete methylation in its presence. Dam methyltransferase is a substrate for the Lon protease. PMID- 12897024 TI - Escherichia coli cells bearing a ribosomal ambiguity mutation in rpsD have a mutator phenotype that correlates with increased mistranslation. AB - Escherichia coli cells bearing certain mutations in rpsD (coding for the 30S ribosomal protein S4) show a ribosomal ambiguity (Ram) phenotype characterized by increased translational error rates. Here we show that spontaneous mutagenesis increases in Ram cells bearing the rpsD14 allele, suggesting that the recently described translational stress-induced mutagenesis pathway is activated in Ram cells. PMID- 12897025 TI - The pst operon of Bacillus subtilis is specifically induced by alkali stress. AB - To cope with a sudden increase in the external pH value to 8.9, Bacillus subtilis cells induce about 80 genes which can be divided into two classes. Most of these genes are members of the sigma(W) regulon, while some are under the control of so far-unknown transcriptional regulators. The genes of the pst operon belong to the second class. Here, we attempted to answer the questions of why and how the genes of this operon are induced. Using transcriptional fusions to two of the five genes of this operon, we confirmed their induction after alkali stress. Furthermore, a Northern blot experiment revealed that the complete operon was alkali inducible, that the transcriptional start site used was identical to that used after phosphate starvation, and that induction was prevented in a phoR background. Most interestingly, increasing the phosphate concentration within the medium prevented alkali induction of the pst operon, and phoA, another member of the PhoRP regulon, did not respond to alkali stress. In the end, we showed that alkali treatment completely prevented phosphate uptake. These results are discussed to explain alkali induction of the pst operon. PMID- 12897026 TI - Conversion of Lactobacillus pentosus D-lactate dehydrogenase to a D hydroxyisocaproate dehydrogenase through a single amino acid replacement. AB - The single amino acid replacement of Tyr52 with Leu drastically increased the activity of Lactobacillus pentosus NAD-dependent D-lactate dehydrogenase toward larger aliphatic or aromatic 2-ketoacid substrates by 3 or 4 orders of magnitude and decreased the activity toward pyruvate by about 30-fold, converting the enzyme into a highly active D-2-hydroxyisocaproate dehydrogenase. PMID- 12897027 TI - A synthetic surfactant based on a poly-Leu SP-C analog and phospholipids: effects on tidal volumes and lung gas volumes in ventilated immature newborn rabbits. AB - Available surfactants for treatment of respiratory distress syndrome in newborn infants are derived from animal lungs, which limits supply and poses a danger of propagating infectious material. Poly-Val-->poly-Leu analogs of surfactant protein (SP)-C can be synthesized in large quantities and exhibit surface activity similar to SP-C. Here, activity of synthetic surfactants containing a poly-Leu SP-C analog (SP-C33) was evaluated in ventilated premature newborn rabbits. Treatment with 2.5 ml/kg body wt of 2% (wt/wt) SP-C33 in 1,2-dipalmitoyl sn-3-glycero phosphoryl choline (DPPC)-1-palmitoyl-2-oleoyl-sn-3-glycero phosphoryl choline (POPC)-1-palmitoyl-2-oleoyl-sn-3-glycero phosphoryl glycerol (POPG), 68:0:31, 68:11:20, or 68:16:15 (wt/wt/wt) suspended at 80 mg/ml gave tidal volumes (Vt) of 20-25 ml/kg body wt, with an insufflation pressure of 25 cmH2O and no positive end-expiratory pressure (PEEP), comparable to the Vt for animals treated with the porcine surfactant Curosurf. Nontreated littermates had a Vt of approximately 2 ml/kg body wt. The Vt for SP-C33 in DPPC-egg phosphatidylglycerol-palmitic acid [68:22:9 (wt/wt/wt)], DPPC-POPG-palmitic acid [68:22:9 (wt/wt/wt)], and DPPC-POPC-POPG [6:2:2 (wt/wt/wt)] was 15-20 ml/kg body wt. Histological examination of lungs from animals treated with SP-C33-based surfactants showed incomplete, usually patchy air expansion of alveolar spaces associated with only mild airway epithelial damage. Lung gas volume after 30 min of mechanical ventilation were more than threefold larger in animals treated with Curosurf than in those receiving SP-C33 in DPPC-POPC-POPG, 68:11:20. This difference could be largely counterbalanced by ventilation with PEEP (3-4 cmH2O). An artificial surfactant based on SP-C33 improves Vt in immature newborn animals ventilated with standardized peak pressure but requires PEEP to build up adequate lung gas volumes. PMID- 12897028 TI - Angiotensin-converting enzyme activity in ovine bronchial vasculature. AB - Angiotensin-converting enzyme (ACE) plays a major role in the metabolism of bradykinin, angiotensin, and neuropeptides, which are all implicated in inflammatory airway diseases. The activity of ACE, which is localized on the luminal surface of endothelial cells (EC), has been well documented in pulmonary EC; however, few data exist regarding the relative activity of ACE in the airway vasculature. Therefore, we measured ACE activity in cultured EC from the sheep bronchial artery and bronchial mucosa (microvascular) and compared it with pulmonary artery EC. The baseline level of total ACE activity (cellular plus secreted) was significantly greater in bronchial microvascular EC (1.24 +/- 0.24 mU/106 cells) compared with bronchial artery EC (0.59 +/- 0.15 mU/106 cells; P < 0.05) and comparable to pulmonary artery EC (1.12 +/- 0.14 mU/106 cells; P > 0.05). Measured ACE activity secreted into culture medium for each cell type was 64-74% of total activity and did not differ among the three EC types (P = 0.17). Hydrocortisone (10 microg/ml; 48-72 h) treatment resulted in a significant increase in ACE activity in bronchial EC. Likewise, TNF-alpha (0.1 ng/ml) treatment markedly increased ACE activity in all cell lysates (P < 0.05). We confirmed the importance of ACE activity in vivo since, at the highest dose of bradykinin studied (10-8 M), bronchial artery pressure at constant flow showed a greater decrease after captopril treatment (36% before vs. 60% after; P = 0.05). These results demonstrate high ACE expression of the bronchial microvasculature and suggest an important regulatory role for ACE in the metabolism of kinin peptides known to contribute to airway pathology. PMID- 12897029 TI - Sleep-disordered breathing, pharyngeal size and soft tissue anatomy in children. AB - We tested the hypothesis that pharyngeal geometry and soft tissue dimensions correlate with the severity of sleep-disordered breathing. Magnetic resonance images of the pharynx were obtained in 18 awake children, 7-12 yr of age, with obstructive apnea-hypopnea index (OAHI) values ranging from 1.81 to 24.2 events/h. Subjects were divided into low-OAHI (n = 9) and high-OAHI (n = 9) groups [2.8 +/- 0.7 and 13.5 +/- 4.9 (SD) P < 0.001]. The OAHI correlated positively with the size of the tonsils (r2 = 0.42, P = 0.024) and soft palate (r2 = 0.33, P = 0.049) and inversely with the volume of the oropharyx (r2 = 0.42, P = 0.038). The narrowest point in the pharyngeal airway was smaller in the high compared with the low-OAHI group (4.4 +/- 1.2 vs. 6.0 +/- 1.3 mm; P = 0.024), and this point was in the retropalatal airway in all but two subjects. The airway cross-sectional area (CSA)-airway length relation showed that the high-OAHI group had a narrower retropapatal airway than the low-OAHI group, particularly in the retropalatal region where the soft palate, adenoids, and tonsils overlap (P = 0.001). The "retropalatal air space," which we defined as the ratio of the retropalatal airway CSA to the CSA of the soft palate, correlated inversely with the OAHI (r2 = 0.49, P = 0.001). We conclude that 7- to 12-yr-old children with a narrow retropalatal air space have significantly more apneas and hypopneas during sleep compared with children with relatively unobstructed retropalatal airways. PMID- 12897030 TI - Modeling kinetics of infused 13NN-saline in acute lung injury. AB - A mathematical model was developed to estimate right-to-left shunt (Fs) and the volume of distribution of 13NN in alveolar gas (VA) and shunt tissue (Vs). The data obtained from this model are complementary to, and obtained simultaneously with, pulmonary functional positron emission tomography (PET). The model describes 13NN kinetics in four compartments: central mixing volume, gas exchanging lung, shunting compartment, and systemic recirculation. To validate the model, five normal prone (NP) and six surfactant-depleted sheep in the supine (LS) and prone (LP) positions were studied under general anesthesia. A central venous bolus of 13NN-labeled saline was injected at the onset of apnea as PET imaging and arterial 13NN sampling were initiated. The model fit the tracer kinetics well (mean r2 = 0.93). Monte Carlo simulations showed that parameters could be accurately identified in the presence of expected experimental noise. Fs derived from the model correlated well with shunt estimates derived from O2 blood concentrations and from PET images. Fs was higher for LS (54 +/- 18%) than for LP (5 +/- 4%) and NP (1 +/- 1%, P < 0.01). VA, as a fraction of PET-measured lung gas volume, was lower for LS (0.18 +/- 0.09) than for LP (0.96 +/- 0.28, P < 0.01), whereas Vs, as a fraction of PET-measured lung tissue volume, was higher for LS (0.46 +/- 0.26) than for LP (0.05 +/- 0.08, P < 0.01). The main conclusions are as follows: 1) the model accurately describes measured arterial 13NN kinetics and provides estimates of Fs, and 2) in this animal model of acute lung injury, the fraction of available gas volume participating in gas exchange is reduced in the supine position. PMID- 12897031 TI - Effect of ischemia and reperfusion without airway occlusion on vascular barrier function in the in vivo mouse lung. AB - Ischemia-reperfusion (I/R) lung injury causes increased vascular permeability and edema. We developed an in vivo murine model of I/R allowing measurement of pulmonary vascular barrier function without airway occlusion. The left pulmonary artery (PA) was occluded with an exteriorized, slipknotted suture in anesthetized C57BL/6J mice. The effect of ischemic time was determined by subjecting mice to 5, 10, or 30 min of left lung ischemia followed by 150 min of reperfusion. The effect of reperfusion time was determined by subjecting mice to 30 min of left lung ischemia followed by 30 or 150 min of reperfusion. Changes in pulmonary vascular barrier function were measured with the Evans blue dye (EBD) technique, dual-isotope radiolabeled albumin (RA), bronchoalveolar lavage (BAL) protein concentration, and wet weight-to-dry weight ratio (WW/DW). Increasing left lung ischemia with constant reperfusion time or increasing left lung reperfusion time after constant ischemic time resulted in significant increases in left lung EBD content at all times compared with both right lung values and sham surgery mice. The effects of left lung ischemia on lung EBD were corroborated by RA but the effects of increasing reperfusion time differed, suggesting binding of EBD to lung tissue. An increase in WW/DW was only detected after 30 min of reperfusion, suggesting edema clearance. BAL protein concentrations were unaffected. We conclude that short periods of I/R, without airway occlusion, increase pulmonary vascular permeability in the in vivo mouse, providing a useful model to study molecular mechanisms of I/R lung injury. PMID- 12897032 TI - Selected Contribution: Identification of differentially expressed genes between young and old rat soleus muscle during recovery from immobilization-induced atrophy. AB - After cessation of hindlimb immobilization, which resulted in a 27-37% loss in soleus mass, the atrophied soleus muscle of young but not old rats regrows to its mass before treatment. We hypothesized that during remobilization the mRNA levels of growth potentiating factor(s) would be present in the soleus muscle of young (3- to 4-mo-old) but absent in old (30- to 31-mo-old) Fischer 344 x Brown Norway rats or that mRNAs for growth inhibitory factor(s) would be absent in young but present in old. Gene expression levels of >24,000 transcripts were determined by using Affymetrix RGU34A-C high-density oligonucleotide microarrays in soleus muscles at 3, 6, 10, and 30 days of remobilization after cessation of a 10-day period of hindlimb immobilization. Each muscle sample was applied to an independent set of arrays. Recovery-related differences were determined by using a three-factor ANOVA with a false discovery rate-adjustment of P = 0.01, which yielded 64 significantly different probe sets. Elfin, amphiregulin, and clusterin mRNAs were selected for further confirmation by real-time PCR. Elfin mRNA levels were less in old than in young rats at 6, 10, and 30 days of remobilization. Amphiregulin expression exhibited a unique spike on the 10th day of successful regrowth in young rats but remained unchanged old. Clusterin mRNA was unchanged in young muscles but was elevated on the 3rd, 6th, and 10th days of recovery in old soleus muscles. The mRNAs identified as differentially expressed between young and old recovery may modulate muscle growth that could highlight new candidate mechanisms to explain the failure of old soleus muscle to recover lost muscle mass. PMID- 12897033 TI - Effects of bronchodilator particle size in asthmatic patients using monodisperse aerosols. AB - Aerosol particle size influences airway drug deposition. Current inhaler devices are inefficient, delivering a heterodisperse distribution of drug particle sizes where, at best, 20% reaches the lungs. Monodisperse aerosols are the appropriate research tools to investigate basic aerosol science concepts within the human airways. We hypothesized that engineering such aerosols of albuterol would identify the ideal bronchodilator particle size, thereby optimizing inhaled therapeutic drug delivery. Eighteen stable mildly to moderately asthmatic patients [mean forced expiratory volume in 1 s (FEV1) 74.3% of predicted] participated in a randomized, double-blind, crossover study design. A spinning top aerosol generator was used to produce monodisperse albuterol aerosols that were 1.5, 3, and 6 microm in size, and also a placebo, which were inhaled at cumulative doses of 10, 20, 40, and 100 microg. Lung function changes and tolerability effects were determined. The larger particles, 6 and 3 microm, were significantly more potent bronchodilators than the 1.5-microm and placebo aerosols for FEV1 and for the forced expiratory flow between exhalation of 25 and 75% of forced vital capacity. A 20-microg dose of the 6- and 3-microm aerosols produced FEV1 bronchodilation comparable to that produced by 200 microg from a metered-dose inhaler. No adverse effects were observed in heart rate and plasma potassium. The data suggest that in mildly to moderately asthmatic patients there is more than one optimal beta2-agonist bronchodilator particle size and that these are larger particles in the higher part of the respirable range. Aerosols delivered in monodisperse form can enable large reductions of the inhaled dose without loss of clinical efficacy. PMID- 12897034 TI - Online recording of ethane traces in human breath via infrared laser spectroscopy. AB - A method is described for rapidly measuring the ethane concentration in exhaled human breath. Ethane is considered a volatile marker for lipid peroxidation. The breath samples are analyzed in real time during single exhalations by means of infrared cavity leak-out spectroscopy. This is an ultrasensitive laser-based method for the analysis of trace gases on the sub-parts per billion level. We demonstrate that this technique is capable of online quantifying of ethane traces in exhaled human breath down to 500 parts per trillion with a time resolution of better than 800 ms. This study includes what we believe to be the first measured expirograms for trace fractions of ethane. The expirograms were recorded after a controlled inhalation exposure to 1 part per million of ethane. The normalized slope of the alveolar plateau was determined, which shows a linear increase over the first breathing cycles and ends in a mean value between 0.21 and 0.39 liter 1. The washout process was observed for a time period of 30 min and was modelled by a threefold exponential decay function, with decay times ranging from 12 to 24, 341 to 481, and 370 to 1770 s. Our analyzer provides a promising noninvasive tool for online monitoring of the oxidative stress status. PMID- 12897035 TI - Tracking cardiac electrical instability by computing interlead heterogeneity of T wave morphology. AB - Oscillations in T-wave morphology, particularly T-wave alternans (TWA), have been fundamentally linked to increased susceptibility to ventricular fibrillation (VF). We investigated whether the escalation in complexity of T-wave oscillations before VF is attributable to increased spatial heterogeneity of repolarization. Peak interlead T-wave heterogeneity (TWH) was measured by second central moment analysis of T-wave morphology in epicardial electrograms in dogs during left anterior descending coronary artery occlusion. TWH differentiated cases in which myocardial ischemia provoked VF from those without VF (563 +/- 56 vs. 139 +/- 36 microV, P < 0.01). In the former group, progressive, significant increases in TWH above preocclusion baseline (70 +/- 8 microV) began at 2.25 min after the start of occlusion and were associated successively with TWA (at 155 +/- 19 microV), T wave multupling (at 386 +/- 100 microV), complex oscillatory T-wave forms (at 560 +/- 76 microV), discordant TWA (at 572 +/- 98 microV), and VF at 4.36 +/- 0.14 min. TWH in precordial ECGs in 12 pigs during angioplasty-balloon-induced myocardial ischemia also discriminated animals that experienced VF (from 90 +/- 14 at baseline to 382 +/- 39 microV, P < 0.05) from those without VF (from 96 +/- 17 at baseline to 199 +/- 61 microV, NS). Ischemia-induced changes in ST segment and T-wave amplitude did not predict VF. Heightened spatial heterogeneity of repolarization, as assessed by second central moment analysis of TWH, underlies TWA and increased risk for ischemia-induced VF. Monitoring spatial TWH from precordial leads could prove useful in stratifying risk for life-threatening arrhythmias. PMID- 12897036 TI - Temporal dynamics of recurrent airway symptoms and cellular random walk. AB - Asthma is a complex chronic inflammatory disease of the small airways that has dramatically increased in prevalence in industrialized countries during the last decades. Risk factors for adult asthma have been related to the complex array of gene-environment interactions and exposure of the immune system to allergens in early childhood. In genetically predisposed subjects, continuous exposure to environmental agents such as allergens or infections can lead to recurrent airway symptoms characterized by recurrent episodes of airway inflammation and bronchoconstriction with clinical symptoms of cough, dyspnea, or wheezing. In this study, we report that the longterm temporal dynamics of recurrent airway symptoms in a population of unselected infants display a complex intermittent pattern and that the distribution of interepisode intervals follows a power law. We interpret the data by using a model of the dynamics of attack episodes in which an attack is triggered by an avalanche of airway constrictions. We map the dynamics of this model to the known problem of a random walk in the presence of an absorbing boundary in which the walker corresponds to the fluctuations in contractile state of airway smooth muscle cells. These findings may provide new insight into the mechanisms of otherwise unexplained symptom episodes. PMID- 12897037 TI - Selected Contribution: Aging impairs nitric oxide and prostacyclin mediation of endothelium-dependent dilation in soleus feed arteries. AB - We tested the hypothesis that endothelium-dependent dilation in soleus muscle feed arteries (SFA) is impaired by aging due to attenuated nitric oxide (NO) mediated vasodilation. SFA were isolated from young (4 mo) and old (24 mo) male Fischer 344 rats and cannulated with two glass micropipettes for examination of endothelium-dependent [flow or acetylcholine (ACh)] and endothelium-independent [sodium nitroprusside (SNP)] vasodilator function. Flow- and ACh-induced dilation was significantly attenuated by age, whereas dilation to SNP was not compromised. To determine the mechanism(s) by which aging affected dilator responses to flow and ACh, dilation was assessed in the presence of Nomega-nitro-L-arginine (L-NNA; to inhibit NO synthase), indomethacin (Indo; to inhibit cyclooxygenase), and L NNA + Indo. In the presence of L-NNA, Indo, or L-NNA + Indo, flow-induced dilation was inhibited in young SFA, resulting in a response to flow that was no longer greater than old SFA. In the presence of L-NNA or Indo, ACh-induced dilation was not significantly inhibited in young or old SFA; however, double blockade with L-NNA + Indo inhibited ACh-induced dilation in young SFA such that the response to ACh was no longer greater than old SFA. Collectively, these data indicate that aging impairs vasodilator responses in SFA by attenuating NO- and prostacyclin-mediated, endothelium-dependent, dilation. PMID- 12897038 TI - Hypothesis: vitamin E complements polyunsaturated fatty acids in essential fatty acid deficiency in cystic fibrosis. AB - While several studies have demonstrated essential fatty acid (EFA) deficiency in plasma and tissue lipids of cystic fibrosis (CF) patients, the reasons for this deficiency are not well established. It is believed that reduced EFA intake, malabsorption of fat, altered desaturase/lipase activity and defective cystic fibrosis transmembrane conductance regulator (CFTR) altering utilisation of EFA in epithelial cells contribute to the development of EFA deficiency in CF. It is likely that increased metabolism of arachidonic acid to eicosanoids such as leukotrienes, thromboxane and prostaglandins may also be a contributing factor. Evidence is presented that elevated oxidative damage to EFA and impaired antioxidant defences, in particular vitamin E, may contribute to the development of EFA deficiency in CF. Furthermore, antioxidant supplementation in CF may improve EFA status. PMID- 12897039 TI - Vitamin E and its role in the prevention of atherosclerosis and carcinogenesis: a review. AB - Epidemiological and experimental studies suggest that antioxidants like vitamin E (alpha-tocopherol) may play an important role in prevention of chronic disease. Several observational surveys have linked populations with a large intake of vitamin E with reduced incidence of heart disease. These observations have been strengthened by the demonstration of strong antioxidant activity by vitamin E in cellular, molecular and animal experiments. These results have highlighted a potential role for vitamin E supplementation in the prevention of chronic disease in humans. Interestingly however, large-scale clinical trials of vitamin E and other antioxidants in preventing specific disease processes (e.g., coronary artery disease) have generated conflicting and mixed outcomes. In this review, the role of vitamin E in the prevention of atherosclerosis and carcinogenesis has been carefully examined with particular emphasis on salient human studies (clinical trials) and their limitations. In addition, pertinent biochemical, physiological and metabolic features of vitamin E have also been incorporated. A list of common natural food sources of vitamin E has been provided. Important in vitro and animal studies related to the antiatherosclerotic and anticarcinogenic actions of vitamin E have been discussed in detail. Finally, the direction of future investigations in primary and secondary prevention of chronic diseases by vitamin E supplementation has been outlined. PMID- 12897040 TI - Predictors of postmenopausal body mass index and waist hip ratio in the oklahoma postmenopausal health disparities study. AB - OBJECTIVE: The goal of the current study has been to examine systematically the respective roles of nutrition, exercise, menopausal weight gain, moderate drinking and smoking as determinants of body mass index (BMI) and waist hip ratio (WHR) in a setting in which the role of race or ethnic group could be simultaneously or individually evaluated as predictors of BMI and WHR. Because the use of estrogen replacement has been reported to affect estimates of body fat mass in postmenopausal women, endocrine factors have also been evaluated. METHODS: The design is cross-sectional with historical prospective elements. The study has a biomedical focus and is not an epidemiologic study. Data are from 649 women recruited into The Postmenopausal Health Disparities Study in Oklahoma. The study population was composed of 649 postmenopausal women: American Indian: 226 (34.9%), Asian: 21 (3.2%), Black: 78 (12.0%), Hispanics: 54 (8.3%) and Whites: 270 (41.6%). Recruitment occurred between 1994 and late 1999 in Oklahoma. RESULTS: In this multi-racial, multi-ethnic study population, there was statistical heterogeneity in all nutrition/dietary and exercise variables as well as in other potential determinants of BMI and WHR. In contrast to the literature available for postmenopausal women in which postmenopausal status, estrogen replacement and race have rarely been taken into account, the results of multi linear regression revealed the following: Significant predictors for BMI, with or without WHR specified, included the neuroendocrine factors, menopausal weight gain, smoking, mean fitness (i.e., difficulty performing physical activities), fat as percent of total calories, moderate drinking and being Asian or Black. When WHR was not included, total calories and socioeconomic status also entered the model. The statistical predictors of WHR in the total study population with BMI in the equation included BMI and the neuroendocrine variables of FSH, E2, but not ERT, as well as the interaction of higher intensity exercise fitness with frequency, socioeconomic status and being American Indian or Asian. When BMI was not included in the model, in addition to the neuroendocrine factors, the interaction of lower intensity exercise fitness with frequency, fat as percent of total calories, age living alone and being American Indian or and Black were significant predictors of WHR. The predictors of both BMI and WHR were found to differ among individual racial and ethnic groups. CONCLUSIONS: Given the role of increased body fat and obesity in disease risk and the substantial differences in life expectancy among the racial and ethnic groups, the findings of this study, particularly in contrast to literature reports, strongly suggest that a whole variety of factors including hormonal status and race need to be considered when examining the role of dietary factors and physical activity in relation to estimates of body fat mass and disease risk. PMID- 12897041 TI - An in vivo study of the antioxidant potentials of a plant food concentrate. AB - OBJECTIVE: A plant food concentrate (PF) is a source of antioxidants. Its influence on antioxidant status has never been studied. The present longitudinal study investigated the antioxidant and lipidemic responses in 15 moderately hypercholesterolemic (>5.2 mmol/L) male subjects to the supplemental intakes of PF. METHODS: The participants underwent a two-week period where any previous supplemental intakes were withdrawn. This was followed by a two-week baseline period at entry (control). The baseline period was followed by taking PF concentrate (8.5 g twice daily) for two weeks followed by a washout period for two weeks. All subjects completed food frequency questionnaires at pre supplemental (baseline) and post-PF period. Fasting heparinized and EDTA blood samples were collected at the end of each period. Erythrocyte superoxide dismutase (SOD), whole blood glutathione peroxidase (GPX) and plasma concentrations of zinc and copper, along with plasma levels of lipids, were determined. RESULTS: The PF supplement contributed significantly to the daily intakes of total dietary fiber. The zinc and copper-dependent SOD but not GPX activity were significantly elevated. The total and LDL-cholesterol concentrations in the plasma were significantly decreased while the ratio of HDL/LDL cholesterol was increased post-PF intake. CONCLUSIONS: These results indicate that the antioxidant and cholesterol status of moderately hypercholesterolemic subjects can be potentially improved with the supplemental intake of PF concentrate. PMID- 12897042 TI - Relation of serum ascorbic acid to Helicobacter pylori serology in US adults: the Third National Health and Nutrition Examination Survey. AB - PURPOSE: To examine the relation between serum ascorbic acid and Helicobacter pylori serology from a probability sample of US adults. SUBJECTS AND METHODS: Data from 6,746 adults (ages 20 to 90 years) enrolled in the Third National Health and Nutrition Examination Survey (NHANES III), 1988-1994 were analyzed. Multiple logistic regression models were examined taking into account sample weights and the complex survey design of NHANES III, and controlling for the effects of potential confounders. Because race appeared to modify the association between serum ascorbic acid and seropositivity to H. pylori, we conducted the analyses stratified by race. RESULTS: A total of 2,189 adults (32%) had a positive serology for H. pylori, and, of these, 1,175 (54%) were positive for the CagA antigen. Among whites, a 0.50 mg/dL increase in serum ascorbic acid level was associated with decreased seroprevalence of H. pylori (Odds Ratio (OR) = 0.89, 95% confidence interval (CI) CI 0.82-0.96, p < 0.01). In analyses that controlled for seroprevalence of H. pylori, a 0.50 mg/dL increase in serum ascorbic acid level among whites was independently associated with a decreased seroprevalence of the pathogenic cagA-positive strain of H. pylori (OR = 0.31, 95% CI 0.12-0.79, p < 0.05). Serum ascorbic acid levels were not significantly associated with H. pylori serology among non-whites (all p > 0.05). CONCLUSIONS: Higher serum levels of ascorbic acid were associated with a decreased seroprevalence of H. pylori and of the pathogenic cagA-positive strain of H. pylori among whites. If these associations are related causally and are not the result of residual confounding by factors such as socioeconomic status, ascorbic acid may affect the risk of H. pylori infection and in turn, the risk for peptic ulcer disease and gastric cancer among white Americans. PMID- 12897043 TI - Hypertension and triglyceride catabolism: implications for the hemodynamic model of the metabolic syndrome. AB - OBJECTIVE: We sought to determine if hypertensive adults have a blunted triglyceride catabolic rate (TG K(2)) and if related hemodynamic and vascular alterations are determinants of TG K(2). METHODS: Fasting levels of insulin, glucose, lipoproteins and plasma catecholamines were measured in 10 normotensive and 10 hypertensive adults. TG K(2) was determined by an intravenous fat tolerance test. Forearm blood flow, maximum forearm blood flow and minimal forearm vascular resistance were determined by strain gauge plethysmography. Vascular compliance and systemic hemodynamics were measured by computerized arterial pulse waveform analysis. RESULTS: Compared to normotensives, hypertensives had a significantly elevated blood pressure (145 +/- 8/94 +/- 11 versus 111 +/- 15/74 +/- 14 mm Hg, p < 0.001), systemic vascular resistance (1695 +/- 441 versus 1172 +/- 430 dynes x sec x cm(-5), p = 0.02) and reduced large vessel compliance (11.7 +/- 3.6 versus 15.1 +/- 3.1 ml/mm Hg x 100, p = 0.04). There were no significant group differences in TG K(2) (3.07 +/- 2.01 versus 2.88 +/- 2.12 mg/dL/min, p = 0.85) or other metabolic and anthropometric variables. TG K(2) was not predicted by the forearm vascular measures or the hemodynamic variables, but was correlated to waist/hip ratio (r = -0.71, p = 0.001), fasting triglycerides (r = -0.64, p = 0.003), and male gender (r = 0.56, p = 0.012). An enhanced TG K(2) was independently predicted by a reduced small vessel compliance (r = -0.61, p = 0.006). CONCLUSIONS: Elevated blood pressure per se and hypertension-related hemodynamic and vascular alterations are not associated with reduced TG K(2) or other metabolic abnormalities. Rather, aspects of the insulin resistance syndrome are closely related to abdominal adiposity. The independent association between small vessel compliance and TG K(2) deserves further investigation. PMID- 12897044 TI - The effect of breakfast type on total daily energy intake and body mass index: results from the Third National Health and Nutrition Examination Survey (NHANES III). AB - OBJECTIVE: The objective of this study was to investigate the relationship between breakfast type, energy intake and body mass index (BMI). We hypothesized not only that breakfast consumption itself is associated with BMI, but that the type of food eaten at breakfast also affects BMI. METHODS: Data from the Third National Health and Nutrition Examination Survey (NHANES III), a large, population-based study conducted in the United States from 1988 to 1994, were analyzed for breakfast type, total daily energy intake, and BMI. The analyzed breakfast categories were "Skippers," "Meat/eggs," "Ready-to-eat cereal (RTEC)," "Cooked cereal," "Breads," "Quick Breads," "Fruits/vegetables," "Dairy," "Fats/sweets," and "Beverages." Analysis of covariance was used to estimate adjusted mean body mass index (BMI) and energy intake (kcal) as dependent variables. Covariates included age, gender, race, smoking, alcohol intake, physical activity and poverty index ratio. RESULTS: Subjects who ate RTEC, Cooked cereal, or Quick Breads for breakfast had significantly lower BMI compared to Skippers and Meat and Egg eaters (p < or = 0.01). Breakfast skippers and fruit/vegetable eaters had the lowest daily energy intake. The Meat and Eggs eaters had the highest daily energy intake and one of the highest BMIs. CONCLUSIONS: This analysis provides evidence that skipping breakfast is not an effective way to manage weight. Eating cereal (ready-to-eat or cooked cereal) or quick breads for breakfast is associated with significantly lower body mass index compared to skipping breakfast or eating meats and/or eggs for breakfast. PMID- 12897045 TI - Changes in serum retinol, alpha-tocopherol, vitamin C, carotenoids, xinc and selenium after micronutrient supplementation during alcohol rehabilitation. AB - OBJECTIVE: To test the effect of a 21-day supplementation with moderate doses of antioxidant nutrients on biochemical indicators of vitamin, carotenoid and trace element levels in alcohol-dependent patients during a program of alcohol rehabilitation. DESIGN: A randomized double-blind trial was performed comparing two groups receiving daily either a combination of micronutrients (beta-carotene: 6 mg, vitamin C: 120 mg, vitamin E: 30 mg, zinc: 20 mg, selenium: 100 micro g) or a placebo. SUBJECTS: 106 alcohol-dependent patients 20 to 60 years of age without severe liver disease, hospitalized for a 21-day rehabilitation program. Measure of Outcome: Vitamin C, retinol, alpha-tocopherol, zeaxanthin/lutein, beta cryptoxanthin, lycopene, alpha- and beta-carotene, zinc and selenium were measured in serum, initially and after supplementation. RESULTS: (1) In the placebo group, after 21 days of rehabilitation, serum concentrations of vitamin C and all five carotenoids significantly increased, whereas retinol and alpha tocopherol concentrations decreased; zinc and selenium levels were unaffected. (2) At the end of the hospital stay, serum indicators were significantly improved in the supplement group as compared to the placebo group for vitamin C, alpha tocopherol, beta-carotene, zinc and selenium; conversely, lycopene changes were higher in the placebo group than in supplement group. (3) Of the serum antioxidants measured at entrance, only vitamin C was significantly depleted in heavy smokers, and, after the supplementation period, vitamin C was efficiently repleted in this later group. CONCLUSION: Our results indicate that a short-term supplementation with physiological doses of antioxidant vitamins, carotenoids and trace elements during alcohol rehabilitation clearly improves micronutrient status indicators. Heavy smokers in particular seem to respond to vitamin C supplementation. PMID- 12897046 TI - Inadequate antioxidant nutrient intake and altered plasma antioxidant status of rheumatoid arthritis patients. AB - OBJECTIVE: Elevated free radical generation in inflamed joints and impaired antioxidant system have been implicated in rheumatoid arthritis (RA). The present study was performed to evaluate dietary nutrient intake and plasma oxidant/antioxidant status in RA patients. METHODS: RA patients (n = 97) and their age, gender-matched controls (n = 97) participated in this cross-sectional case-control study. Nutrient intake was estimated using a semi-quantitative food frequency questionnaire. Twenty subjects from each group provided blood samples, and plasma concentrations of alpha-tocopherol and malondiadehyde (MDA) were measured. Also, plasma activities of superoxide dismutase (SOD) and glutathione peroxidase (GPx) were measured. RESULTS: The mean calorie intake of RA patients was lower than that of the healthy controls. Energy-adjusted intake of fat, vitamin A and beta-carotene were significantly lower in patients than those of the control subjects. RA patients had a decreased mean plasma alpha-tocopherol level. The activity of plasma SOD and GPx in patients was significantly lower than that in control subjects. CONCLUSION: These results suggest proper antioxidant nutrient intake management may reduce free radical generation and improve antioxidant status in RA patients. PMID- 12897047 TI - Antioxidant effects of zinc supplementation in Tunisians with type 2 diabetes mellitus. AB - OBJECTIVE: To determine the effects of zinc (Zn) supplementation on oxidative stress in persons with type 2 diabetes mellitus (type 2 DM). DESIGN: Tunisian adult subjects with HbA1c >7.5% were supplemented for six months with 30 mg/day of Zn as Zn gluconate or placebo. The effects of supplementation on plasma zinc (Zn), copper (Cu), urinary Zn, plasma thiobarbituric acid reactive substances (TBARS), Cu-Zn superoxide dismutase (SOD) and glutathione peroxidase activities (GPX) in red blood cells, blood lipids and lipoproteins, HbA1c and fasting glucose were measured at the beginning of the study and after three and six months. RESULTS: At the beginning of the study, more than 30% of the subjects exhibited plasma Zn values less than the normal minimum of 10.7 micro mol/L, whereas levels of plasma Cu and antioxidant RBC Cu-Zn SOD and GPx enzyme activities were in the normal ranges. Oxidative stress, monitored by plasma TBARS, was increased in individuals with diabetes compared with healthy Tunisian subjects (3.32 +/- 0.05 micro mol/L vs. 2.08 +/- 0.04 micro mol/L) and an inverse correlation was found between Zn plasma levels and plasma TBARS. After three and six months of Zn supplementation, all of the subjects exhibited plasma Zn values greater than 10.7 micro mol/L. There was a decrease of plasma TBARS in Zn supplemented group after six months (15%) with no significant changes in the placebo group. Supplementation did not alter significantly HbA1c nor glucose homeostasis. No adverse effects of Zn supplementation were observed on Cu status or HDL cholesterol. CONCLUSIONS: These data suggest the potential beneficial antioxidant effects of Zn supplementation in persons with type 2 DM. These results are particularly important in light of the deleterious consequences of oxidative stress in persons with diabetes. PMID- 12897048 TI - Protective effects of flax meal against hypercholesterolemia and hypertriglyceridemia in rats. PMID- 12897049 TI - Heterochromatin: stable and unstable invasions at home and abroad. PMID- 12897050 TI - The pims and outs of survival signaling: role for the Pim-2 protein kinase in the suppression of apoptosis by cytokines. PMID- 12897051 TI - Differential DNA affinity specifies roles for the origin recognition complex in budding yeast heterochromatin. AB - The origin recognition complex (ORC) marks chromosomal positions as replication origins and is essential for replication initiation. At a few loci, the ORC functions in heterochromatin formation. We show that the ORC's two roles at the heterochromatic HMRa locus in Saccharomyces cerevisiae were regulated by differences in the ORC's interaction with its target site. At HMRa, a strong ORC DNA interaction inhibited and delayed replication initiation but promoted heterochromatin formation, whereas a weak ORC-DNA interaction allowed for increased and earlier replication initiation but reduced heterochromatin formation. Therefore, the ORC's interaction with its target site could modulate ORC activity within a heterochromatin domain in vivo. PMID- 12897052 TI - Structural basis for specific binding of Polycomb chromodomain to histone H3 methylated at Lys 27. AB - The chromodomain of Drosophila Polycomb protein is essential for maintaining the silencing state of homeotic genes during development. Recent studies suggest that Polycomb mediates the assembly of repressive higher-order chromatin structures in conjunction with the methylation of Lys 27 of histone H3 by a Polycomb group repressor complex. A similar mechanism in heterochromatin assembly is mediated by HP1, a chromodomain protein that binds to histone H3 methylated at Lys 9. To understand the molecular mechanism of the methyl-Lys 27 histone code recognition, we have determined a 1.4-A-resolution structure of the chromodomain of Polycomb in complex with a histone H3 peptide trimethylated at Lys 27. The structure reveals a conserved mode of methyl-lysine binding and identifies Polycomb specific interactions with histone H3. The structure also reveals a dPC dimer in the crystal lattice that is mediated by residues specifically conserved in the Polycomb family of chromodomains. The dimerization of dPC can effectively account for the histone-binding specificity and provides new mechanistic insights into the function of Polycomb. We propose that self-association is functionally important for Polycomb. PMID- 12897053 TI - Endothelial PDGF-B retention is required for proper investment of pericytes in the microvessel wall. AB - Several platelet-derived growth factor (PDGF) and vascular endothelial growth factor (VEGF) family members display C-terminal protein motifs that confer retention of the secreted factors within the pericellular space. To address the role of PDGF-B retention in vivo, we deleted the retention motif by gene targeting in mice. This resulted in defective investment of pericytes in the microvessel wall and delayed formation of the renal glomerulus mesangium. Long term effects of lack of PDGF-B retention included severe retinal deterioration, glomerulosclerosis, and proteinuria. We conclude that retention of PDGF-B in microvessels is essential for proper recruitment and organization of pericytes and for renal and retinal function in adult mice. PMID- 12897054 TI - Molecular basis for the discrimination of repressive methyl-lysine marks in histone H3 by Polycomb and HP1 chromodomains. AB - On the histone H3 tail, Lys 9 and Lys 27 are both methylation sites associated with epigenetic repression, and reside within a highly related sequence motif ARKS. Here we show that the chromodomain proteins Polycomb (Pc) and HP1 (heterochromatin protein 1) are highly discriminatory for binding to these sites in vivo and in vitro. In Drosophila S2 cells, and on polytene chromosomes, methyl Lys 27 and Pc are both excluded from areas that are enriched in methyl-Lys 9 and HP1. Swapping of the chromodomain regions of Pc and HP1 is sufficient for switching the nuclear localization patterns of these factors, indicating a role for their chromodomains in both target site binding and discrimination. To better understand the molecular basis for the selection of methyl-lysine binding sites, we solved the 1.8 A structure of the Pc chromodomain in complex with a H3 peptide bearing trimethyl-Lys 27, and compared it with our previously determined structure of the HP1 chromodomain in complex with a H3 peptide bearing trimethyl Lys 9. The Pc chromodomain distinguishes its methylation target on the H3 tail via an extended recognition groove that binds five additional residues preceding the ARKS motif. PMID- 12897055 TI - Sequence-independent DNA binding and replication initiation by the human origin recognition complex. AB - We report that a highly purified human origin recognition complex (HsORC) has intrinsic DNA-binding activity, and that this activity is modestly stimulated by ATP. HsORC binds preferentially to synthetic AT-rich polydeoxynucleotides, but does not effectively discriminate between natural DNA fragments that contain known human origins and control fragments. The complex fully restores DNA replication to ORC-depleted Xenopus egg extracts, providing strong evidence for its initiator function. Strikingly, HsORC stimulates initiation from any DNA sequence, and it does not preferentially replicate DNA containing human origin sequences. These data provide a biochemical explanation for the observation that in metazoans, initiation of DNA replication often occurs in a seemingly random pattern, and they have important implications for the nature of human origins of DNA replication. PMID- 12897056 TI - A conserved structural motif reveals the essential transcriptional repression function of Spen proteins and their role in developmental signaling. AB - Spen proteins regulate the expression of key transcriptional effectors in diverse signaling pathways. They are large proteins characterized by N-terminal RNA binding motifs and a highly conserved C-terminal SPOC domain. The specific biological role of the SPOC domain (Spen paralog and ortholog C-terminal domain), and hence, the common function of Spen proteins, has been unclear to date. The Spen protein, SHARP (SMRT/HDAC1-associated repressor protein), was identified as a component of transcriptional repression complexes in both nuclear receptor and Notch/RBP-Jkappa signaling pathways. We have determined the 1.8 A crystal structure of the SPOC domain from SHARP. This structure shows that essentially all of the conserved surface residues map to a positively charged patch. Structure-based mutational analysis indicates that this conserved region is responsible for the interaction between SHARP and the universal transcriptional corepressor SMRT/NCoR (silencing mediator for retinoid and thyroid receptors/nuclear receptor corepressor. We demonstrate that this interaction involves a highly conserved acidic motif at the C terminus of SMRT/NCoR. These findings suggest that the conserved function of the SPOC domain is to mediate interaction with SMRT/NCoR corepressors, and that Spen proteins play an essential role in the repression complex. PMID- 12897057 TI - BMAL1-dependent circadian oscillation of nuclear CLOCK: posttranslational events induced by dimerization of transcriptional activators of the mammalian clock system. AB - Mammalian CLOCK and BMAL1 are two members of bHLH-PAS-containing family of transcription factors that represent the positive elements of circadian autoregulatory feedback loop. In the form of a heterodimer, they drive transcription from E-box enhancer elements in the promoters of responsive genes. We have examined abundance, posttranslational modifications, cellular localization of endogenous and ectopically expressed CLOCK and BMAL1 proteins. Nuclear/cytoplasm distribution of CLOCK was found to be under circadian regulation. Analysis of subcellular localization of CLOCK in embryo fibroblasts of mice carrying different germ-line circadian mutations showed that circadian regulation of nuclear accumulation of CLOCK is BMAL1-dependent. Formation of CLOCK/BMAL1 complex following ectopic coexpression of both proteins is followed by their codependent phosphorylation, which is tightly coupled to CLOCK nuclear translocation and degradation. This binding-dependent coregulation is specific for CLOCK/BMAL1 interaction, as no other PAS domain protein that can form a complex with either CLOCK or BMAL1 was able to induce similar effects. Importantly, all posttranslational events described in our study are coupled with active transactivation complex formation, which argues for their significant functional role. Altogether, these results provide evidence for an additional level of circadian system control, which is based on regulation of transcriptional activity or/and availability of CLOCK/BMAL1 complex. PMID- 12897058 TI - Ser-256 phosphorylation dynamics of Aquaporin 2 during maturation from the ER to the vesicular compartment in renal cells. AB - Aquaporin 2 (AQP2) phosphorylation at Ser-256 by protein kinase A (PKA) is a key signal for vasopressin-stimulated AQP2 insertion into the plasma membrane in renal cells. This study underscores the possible role of phosphorylation at Ser 256 in regulating AQP2 maturation. AQP2-transfected renal CD8 cells were incubated with brefeldin A (BFA) to accumulate newly synthesized AQP2 in the endoplasmic reticulum (ER), and AQP2 flow from ER to the vesicular compartment was analyzed after BFA washout. We found that a) in the ER, AQP2 is weakly phosphorylated; b) the amount of phosphorylated AQP2 (p-AQP2) at Ser-256 increased significantly during transit in the Golgi, even in the presence of the PKA inhibitor H89; and c) AQP2 transport from the Golgi to the vasopressin regulated vesicular compartment occurred with a concomitant decrease in p-AQP2 at Ser-256. These results support the hypothesis that AQP2 transition in the Golgi apparatus is associated with a PKA-independent increase in AQP2 phosphorylation at Ser-256. Conversely, impaired constitutive phosphorylation in a Golgi associated compartment occurring in cells expressing mutated S256A-AQP2 or E258K AQP2 causes phosphorylation-defective AQP2 routing to lysosomes. This result might explain the molecular basis of the dominant form of nephrogenic diabetes insipidus caused by the mutation E258K-AQP2, in which the phenotype is caused by an impaired routing of AQP2. PMID- 12897059 TI - Dual mechanisms of green tea extract (EGCG)-induced cell survival in human epidermal keratinocytes. AB - Beneficial effects attributed to green tea, such as its anticancer and antioxidant properties, may be mediated by (-)-epigallocatechin-3-gallate (EGCG). In this study, the effects of EGCG on cell proliferation and UV-induced apoptosis were investigated in normal epidermal keratinocytes. When topically applied to aged human skin, EGCG stimulated the proliferation of epidermal keratinocytes, which increased the epidermal thickness. In addition, this topical application also inhibited the UV-induced apoptosis of epidermal keratinocytes. EGCG was found to increase the phosphorylation of Bad protein at the Ser112 and Ser136. Moreover, EGCG-induced Erk phosphorylation was found to be critical for the phosphorylation of Ser112 in Bad protein, and the EGCG-induced activation of the Akt pathway was found to be involved in the phosphorylation of Ser136. Furthermore, EGCG increased Bcl-2 expression but decreased Bax expression, causing an increase in the Bcl-2-to-Bax ratio. In addition, we demonstrate the differential growth inhibitory effects of EGCG on cancer cells. In conclusion, this study demonstrates that EGCG promotes keratinocyte survival and inhibits the UV-induced apoptosis via two mechanisms: by phosphorylating Ser112 and Ser136 of Bad protein through Erk and Akt pathways, respectively, and by increasing the Bcl 2-to-Bax ratio. Moreover, these two proposed mechanisms of EGCG-induced cell proliferation may differ kinetically to promote keratinocyte survival. PMID- 12897060 TI - PPARgamma ligands induce prostaglandin production in vascular smooth muscle cells: indomethacin acts as a peroxisome proliferator-activated receptor-gamma antagonist. AB - Peroxisome proliferator-activated receptor (PPAR)gamma and inducible cyclooxygenase-2 (COX-2) are expressed in atherosclerotic lesions, particularly in the intimal monocytic and vascular smooth muscle cells. We have therefore studied the interaction between PPARgamma and inducible cyclo-oxygenase (COX-2) in rat aortic vascular smooth muscle cells (RASMC)s. The synthetic PPARgamma ligand rosiglitazone induced prostaglandin (PG) release from RASMCs, including that of PGD2, the precursor of the putative endogenous PPARgamma ligand 15-deoxy Delta12,14-prostaglandin J2. Moreover, rosiglitazone both synergized with IL 1beta to further induce prostaglandin release and affected the expression of phospholipase A2 and COX-2. Rosiglitazone-induced prostaglandin release was inhibited by the PPARgamma partial agonist GW0072 and the PPARgamma antagonist GW9662. Rosiglitazone also induced RASMC apoptosis, an effect not explained as an autocrine effect of the induced-prostanoids, but on arachidonic acid release, as cell death was unaffected by either the nonselective COX inhibitor piroxicam or the selective COX-2 inhibitor DFP, but by inhibitors of either secretory or cytosolic phospholipase A2. In contrast, indomethacin, an alternative inhibitor of cyclooxygenase activity, inhibited both rosiglitazone-induced cell death, and rosiglitazone-induced PPAR reporter gene activation. PMID- 12897061 TI - Impaired adrenal catecholamine system function in mice with deficiency of the ascorbic acid transporter (SVCT2). AB - Ascorbic acid (vitamin C) is a cofactor required in catecholamine synthesis for conversion of dopamine to norepinephrine by dopamine beta-hydroxylase. Mutant mice lacking the plasma membrane ascorbic acid transporter (SVCT2) have severely reduced tissue levels of ascorbic acid and die after birth. We therefore investigated whether these mice might have impaired synthesis of catecholamines. Levels of catecholamines in brain were unaffected by SVCT2 deficiency. In heart, the only evidence for impaired dopamine beta-hydroxylase activity was a twofold increase in tissue dopamine. An influence of the deficiency on tissue catecholamines was most prominent in the adrenals where norepinephrine was decreased by 50% and epinephrine, by 81%. On the ultrastructural level, adrenal chromaffin cells in SVCT2 null mice showed depletion of catecholamine storage vesicles, increased amounts of rough endoplasmic reticulum, signs of apoptosis, and increased glycogen storage. Decreased plasma levels of corticosterone indicated additional effects of the deficiency on adrenal cortical function. These data show that deranged catecholamine system function in SVCT2 null mice is largely restricted to the adrenal medulla and cannot account for the lethality in these animals. The data, however, establish a crucial role for ascorbic acid in adrenal chromaffin cell function. PMID- 12897062 TI - Up-regulation of cardiac nitric oxide synthase 1-derived nitric oxide after myocardial infarction in senescent rats. AB - Nitric oxide (NO) has been implicated in the development of heart failure, although the source, significance, and functional role of the different NO synthase (NOS) isoforms in this pathology are controversial. The presence of a neuronal-type NOS isoform (NOS1) in the cardiac sarcoplasmic reticulum has been recently discovered, leading to the hypothesis that NOS1-derived NO may notably alter myocardial inotropy. However, the regulation and role(s) of NOS1 in cardiac diseases remain to be determined. Using an experimental model of myocardial infarction (MI) in senescent rats, we demonstrated a significant increase in cardiac NOS1 expression and activity in MI, coupled with the translocation of this enzyme to the sarcolemma through interactions with caveolin-3. The enhanced NOS1 activity counteracts the decrease in cardiac NOS3 expression and activity observed in heart failure. We demonstrated an increased interaction between NOS1 and its regulatory protein HSP90 in post-MI hearts, a potential mechanism for the higher NOS1 activity in this setting. Finally, preferential in vivo inhibition of NOS1 activity enhanced basal post-MI left ventricular dysfunction in senescent rats. These results provide the first evidence that increased NOS1-derived NO production may play a significant role in the autocrine regulation of myocardial contractility after MI in aging rats. PMID- 12897063 TI - Angiogenic role of adrenomedullin through activation of Akt, mitogen-activated protein kinase, and focal adhesion kinase in endothelial cells. AB - Adrenomedullin (AM) is a multifunctional peptide in human pheochromocytoma. To evaluate whether AM could be an angiogenic factor, we examined its effect on kinases and angiogenic processes. AM induced tyrosine phosphorylation of Akt and mitogen-activated protein kinase (MAPK)/extracellular signal-regulated kinase1/2 (ERK1/2) by using distinct signaling pathways in human umbilical vein endothelial cells (HUVECs). AM also phosphorylated focal adhesion kinase, and phosphatidylinositol 3'-kinase inhibitor inhibited AM-induced focal adhesion kinase phosphorylation. Pretreatment with high concentrations of AM22-52, a putative AM receptor antagonist, partially suppressed AM-induced phosphorylation of Akt, ERK1/2, and focal adhesion kinase. AM and vascular endothelial growth factor produced increases in DNA synthesis and migration in HUVECs. AM induced tube formation in HUVECs, and its effect was inhibited by pretreatment with phosphatidylinositol 3'-kinase inhibitor or ERK1/2 inhibitor. AM induced sprouting in porcine pulmonary arterial endothelial cells and promoted neovessel formation in a mouse Matrigel plug assay. Inhibitors of phosphatidylinositol 3' kinase and ERK1/2 inhibited AM-induced endothelial sprouting in vitro and angiogenesis in vivo. AM exerts angiogenic activity through activation of Akt, MAPK, and focal adhesion kinase in endothelial cells. PMID- 12897064 TI - Neutrophil C5a receptor and the outcome in a rat model of sepsis. AB - Complement fragment 5a (C5a)-C5a receptor (C5aR) signaling plays an essential role in neutrophil innate immunity. Blockade of either the ligand or the receptor improves survival rates in experimental sepsis. In the current study, sepsis was induced in rats by cecal ligation/puncture. Early in sepsis C5aR content on neutrophils significantly dropped, reached the nadir at 24 h after onset of sepsis, and progressively elevated thereafter. Western-blot, RT-PCR, and confocal microscopy analyses revealed that the loss and re-expression of C5aR during sepsis might be due, at least in part, to the receptor internalization and reconstitution. The reduction and reconstitution of C5aR correlate with the loss and restoration of innate immune functions of blood neutrophils (chemotaxis and reactive oxygen species production), respectively. Quantitative measurements of C5aR on blood neutrophils are highly predictive of survival or death during sepsis. These data suggest that neutrophil C5aR content represents an essential component of an efficient defense system in sepsis and may serve as a prognostic marker for the outcome. PMID- 12897065 TI - Flavonoid wogonin from medicinal herb is neuroprotective by inhibiting inflammatory activation of microglia. AB - Wogonin (5,7-dihydroxy-8-methoxyflavone), a flavonoid originated from the root of a medicinal herb Scutellaria baicalensis Georgi, has been previously shown to have anti-inflammatory activities in various cell types including macrophages. In this work, we have found that wogonin is a potent neuroprotector from natural source. Wogonin inhibited inflammatory activation of cultured brain microglia by diminishing lipopolysaccharide-induced tumor necrosis factor-alpha (TNF-alpha), interleukin-1beta, and nitric oxide (NO) production. Wogonin inhibited NO production by suppressing inducible NO synthase (iNOS) induction and NF-kappaB activation in microglia. Inhibition of inflammatory activation of microglia by wogonin led to the reduction in microglial cytotoxicity toward cocultured PC12 cells, supporting a neuroprotective role for wogonin in vitro. The neuroprotective effect of wogonin was further demonstrated in vivo using two experimental brain injury models; transient global ischemia by four-vessel occlusion and excitotoxic injury by systemic kainate injection. In both animal models, wogonin conferred neuroprotection by attenuating the death of hippocampal neurons, and the neuroprotective effect was associated with inhibition of the inflammatory activation of microglia. Hippocampal induction of inflammatory mediators such as iNOS and TNF-alpha was reduced by wogonin in the global ischemia model, and microglial activation was markedly down-regulated by wogonin in the kainate injection model as judged by microglia-specific isolectin B4 staining. Taken together, our results indicate that wogonin exerts its neuroprotective effect by inhibiting microglial activation, which is a critical component of pathogenic inflammatory responses in neurodegenerative diseases. The current study emphasizes the importance of medicinal herbs and their constituents as an invaluable source for the development of novel neuroprotective drugs. PMID- 12897066 TI - Type 1 plasminogen activator inhibitor deficiency aggravates the course of experimental glomerulonephritis through overactivation of transforming growth factor beta. AB - Type 1 plasminogen activator inhibitor (PAI-1) is the primary inhibitor of tissue type plasminogen activator (tPA) and urokinase-type plasminogen activator (uPA). Whereas PAI-1 is not expressed in normal kidneys, it is strongly induced in glomerular diseases and thus could promote the local accumulation of fibrin. To study the role of PAI-1 in the development of inflammatory glomerular injury, passive antiglomerular basement membrane (GBM) glomerulonephritis (GN) was induced in PAI-1 knockout mice and in wild-type mice of the same genetic background. Unexpectedly, PAI-1 deficiency was associated with an early and severe exacerbation of glomerular injury: Infiltration by CD4 T cells, proportion of fibrinous crescents, and renal function impairment were significantly more pronounced in PAI-1 -/- mice. Interestingly, activation of transforming growth factor (TGF)- beta, which is known to be dependent on the PA/plasmin system in vitro, was dramatically enhanced in the kidneys in the absence of PAI-1. Moreover, administration of neutralizing antibodies against TGF-beta significantly attenuated the disease in PAI-1 -/- mice. This suggests that the poor outcome of GN in PAI-1 -/- mice is consecutive to an uncontrolled activation of TGF-beta and confers PAI-1 with a new, immunomodulatory role. PMID- 12897067 TI - The alpha7 nicotinic acetylcholine receptor in schizophrenia: decreased mRNA levels in peripheral blood lymphocytes. AB - Recent studies have suggested that the alpha7 nicotinic acetylcholine receptor (alpha7 AChR) may play a role in the pathogenesis of schizophrenia. In search for peripheral biological markers for schizophrenia we have investigated alpha7 mRNA levels in peripheral blood lymphocytes (PBLs) of schizophrenic patients and healthy controls. Peripheral blood samples were collected from medicated and non medicated (drug naive) schizophrenic patients as well as from healthy (non mentally ill) smokers and non-smokers. RNA was prepared from isolated lymphocytes. Polymerase chain reaction products specific for human alpha7 AChR were quantified by densitometry using Scion image-analysis (shared NIH software). We observed a significant decrease of alpha7 mRNA levels on PBLs of schizophrenic patients compared with controls. The decrease in alpha7 mRNA levels was not a result of medication management, because non-medicated schizophrenic patients displayed the same level of reduction in alpha7 mRNA as did patients receiving medication. In addition, we exclude the possibility that the observed decrease in alpha7 mRNA levels resulted from nicotine consumption in smoking, because healthy smokers exhibited the same levels of alpha7 mRNA as non-smokers. We propose that alpha7 AChR may be involved in the pathophysiology of the disease and may serve as a reliable peripheral biological marker in schizophrenia. PMID- 12897068 TI - Critical role of microglial NADPH oxidase-derived free radicals in the in vitro MPTP model of Parkinson's disease. AB - 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) damages dopaminergic neurons as seen in Parkinson's disease. Although increasing evidence suggests an involvement of glia in MPTP neurotoxicity, the nature of this involvement remains unclear. Exploiting the advantage of cell culture systems, we demonstrated that microglia, but not astroglia, significantly enhanced the progression of MPTP induced dopaminergic neurodegeneration. Characterization of the temporal relationship between neurodegeneration and microglial activation demonstrates that reactive microgliosis resulting from MPTP-initiated neuronal injury, but not direct activation, underlies the microglia-enhanced MPTP neurotoxicity. Mechanistically, through the release of NADPH oxidase-derived reactive oxygen species, microglia contribute to the progressive neuronal damage. Among the factors measured, the production of extracellular superoxide was the most prominent. NADPH oxidase inhibitor, apocynin, attenuated MPTP-induced dopaminergic neurodegeneration only in the presence of glia. More importantly, dopaminergic neurons from mice lacking NADPH oxidase, a key enzyme for superoxide production in immune cells, are significantly more resistant to MPTP neurotoxicity than those from wild-type controls, and microglia dictate the resistance. This study demonstrates that reactive microgliosis triggered by MPTP induced neuronal injury and NADPH oxidase-mediated superoxide production in microglia constitute an integral component of MPTP neurotoxicity. This study also suggests that NADPH oxidase may be a promising target for therapeutic interventions in Parkinson's disease. PMID- 12897069 TI - Heat shock factor functions at the convergence of the stress response and developmental pathways in Caenorhabditis elegans. AB - Heat shock factor (HSF) is best characterized as the transcriptional regulator of heat shock protein genes, required by all cells to survive periods of stress. Recent evidence suggests that HSF also functions to regulate the expression of genes involved in growth and development under normal physiological conditions. In this study, we used RNA interference (RNAi) assays to investigate the role of HSF in Caenorhabditis elegans. Exposure of wild-type worms to hsf dsRNAi constructs caused a temperature-sensitive developmental arrest at the L2/L3 stage. At normal growth temperatures, hsf(RNAi) worms that developed to adults were small and scrawny, largely infertile, and showed a significant reduction in life span. These results demonstrate that HSF is required for normal postembryonic development under physiological conditions. Following heat shock, hsf(RNAi) worms were thermosensitive and displayed a significant reduction of hsp16 expression. When hsf(RNAi) was carried out in various dauer-constitutive mutant backgrounds, a dramatic reversal of dauer formation was observed, indicating that HSF is also required in the dauer pathway. In its natural habitat of the soil, where C. elegans is exposed to a constantly fluctuating environment; the ability to integrate the stress response with development may be an essential element of its ecology. PMID- 12897070 TI - Stability of the nuclear protein turnover during cellular senescence of human fibroblasts. AB - The accumulation of oxidized proteins is one of the highlights of age-related changes of cellular metabolism and happens at least partially as a result of a decline in the activity of intracellular proteases (e.g., the proteasome). Because the proteasome is located in numerous cellular compartments, we tested whether and to which extent the proteasome and the protein turnover changes in the cytosolic compartment and in the nucleus of proliferating fibroblasts. We demonstrated that the activity of the proteasomal system declines during proliferative senescence of human fibroblasts in the cytosol dramatically, whereas it is stable within the nucleus. It could be demonstrated in both compartments that an accumulation of oxidized proteins occurs. After oxidative stress, a short timed activation of the proteasomal system in the nucleus occurs. This activation was accompanied by an increase in the protein turnover in response to oxidative stress, which was also present in the nucleus of senescent cells. Taking into account that the nuclear/cytosol ratio of the proteasome content declines during proliferative senescence, we postulated that the senescence-related changes in the cytosolic proteasomal system are more pronounced and that the nuclear proteasomal system is only marginally affected by the senescence process. PMID- 12897071 TI - A region directly following the second transmembrane domain in gamma ENaC is required for normal channel gating. AB - We used a yeast one-hybrid complementation screen to identify regions within the cytosolic tails of the mouse alpha, beta, and gamma epithelial Na+ channel (ENaC) important to protein-protein and/or protein-lipid interactions at the plasma membrane. The cytosolic COOH terminus of alphaENaC contained a strongly interactive domain just distal to the second transmembrane region (TM2) between Met610 and Val632. Likewise, gammaENaC contained such a domain just distal to TM2 spanning Gln573-Pro600. Interactive domains were also localized within Met1-Gln54 and the last 17 residues of alpha- and betaENaC, respectively. Confocal images of Chinese hamster ovary cells transfected with enhanced green fluorescent fusion proteins of the cytosolic tails of mENaC subunits were consistent with results in yeast. Fusion proteins of the NH2 terminus of alphaENaC and the COOH termini of all three subunits co-localized with a plasma membrane marker. The functional importance of the membrane interactive domain in the COOH terminus of gammaENaC was established with whole-cell patch clamp experiments of wild type (alpha, beta, and gamma) and mutant (alpha, beta, and gammadeltaQ573-P600) mENaC reconstituted in Chinese hamster ovary cells. Mutant channels had about 13% of the activity of wild type channels with 0.33 +/- 0.14 versus 2.5 +/- 0.80 nA of amiloridesensitive inward current at -80 mV. Single channel analysis of recombinant channels demonstrated that mutant channels had a decrease in Po with 0.16 +/- 0.03 versus 0.67 +/- 0.07 for wild type. Mutant gammaENaC associated normally with the other two subunits in co-immunoprecipitation studies and localized to the plasma membrane in membrane labeling experiments and when visualized with evanescent-field fluorescence microscopy. Similar to deletion of Gln573-Pro600, deletion of Gln573-Arg583 but not Thr584-Pro600 decreased ENaC activity. The current results demonstrate that residues within Gln573-Arg583 of gammaENaC are necessary for normal channel gating. PMID- 12897072 TI - Xenopus Drf1, a regulator of Cdc7, displays checkpoint-dependent accumulation on chromatin during an S-phase arrest. AB - We have cloned a Xenopus Dbf4-related factor named Drf1 and characterized this protein by using Xenopus egg extracts. Drf1 forms an active complex with the kinase Cdc7. However, most of the Cdc7 in egg extracts is not associated with Drf1, which raises the possibility that some or all of the remaining Cdc7 is bound to another Dbf4-related protein. Immunodepletion of Drf1 does not prevent DNA replication in egg extracts. Consistent with this observation, Cdc45 can still associate with chromatin in Drf1-depleted extracts, albeit at significantly reduced levels. Nonetheless, Drf1 displays highly regulated binding to replicating chromatin. Treatment of egg extracts with aphidicolin results in a substantial accumulation of Drf1 on chromatin. This accumulation is blocked by addition of caffeine and by immunodepletion of either ATR or Claspin. These observations suggest that the increased binding of Drf1 to aphidicolin-treated chromatin is an active process that is mediated by a caffeine-sensitive checkpoint pathway containing ATR and Claspin. Abrogation of this pathway also leads to a large increase in the binding of Cdc45 to chromatin. This increase is substantially reduced in the absence of Drf1, which suggests that regulation of Drf1 might be involved in the suppression of Cdc45 loading during replication arrest. We also provide evidence that elimination of this checkpoint causes resumed initiation of DNA replication in both Xenopus tissue culture cells and egg extracts. Taken together, these observations argue that Drf1 is regulated by an intra-S-phase checkpoint mechanism that down-regulates the loading of Cdc45 onto chromatin containing DNA replication blocks. PMID- 12897073 TI - Angiotensin II, the immune system and renal diseases: another road for RAS? PMID- 12897074 TI - The emerging role of VEGF in diabetic kidney disease. PMID- 12897075 TI - The radical treatment of paraprotein disorders affecting the kidney. PMID- 12897076 TI - Oligonephronia, primary hypertension and renal disease: 'is the child father to the man?'. PMID- 12897077 TI - Oxidative stress: does it play a role in the genesis of essential hypertension and hypertension of uraemia? PMID- 12897078 TI - Ageing, hypertension and the kidney: new data on an old problem. PMID- 12897079 TI - The advantage of a uniform terminology and staging system for chronic kidney disease (CKD). PMID- 12897080 TI - Has the time come to use antioxidant therapy in uraemic patients? PMID- 12897081 TI - When should pregnant women with an elevated blood pressure be treated? PMID- 12897082 TI - Cyclosporin increases the density of angiotensin II subtype 1 (AT1) receptors in mouse medullary thick ascending limb cells. AB - BACKGROUND: Cyclosporin A (CsA), a potent immunosuppressive agent, can be nephrotoxic. Because clinical studies have suggested that the intrarenal renin angiotensin system may be involved in the mechanism responsible for CsA nephrotoxicity, we have analysed the effects of CsA on angiotensin II (Ang II) receptors in medullary thick ascending limb (mTAL) cells known to be sensitive to the action of CsA. METHODS: Experiments were carried out on subcultured mouse mTAL cells. The expression of mRNA of Ang II subtype 1 and 2 (AT(1) and AT(2)) receptors was investigated using reverse transcription-polymerase chain reaction (RT-PCR). [(3)H]Ang II was used for radioligand and binding studies. Fluorimetric recordings using the fluorescent dye fura-2/AM were performed to determine the effect of CsA on the intracellular calcium ([Ca(2+)]i) content of untreated and Ang II-treated mTAL cells. RESULTS: Subcultured mTAL cells expressed AT(1) and AT(2) Ang II receptor mRNAs, and binding studies revealed that the AT(1) receptors were the predominant Ang II receptor subtype ( approximately 90%) in mTAL cells. CsA (100 ng/ml, 24 h) increased (1.7-fold) the number of Ang II receptors (untreated, 315.8; +CsA, 543.6 fmol/mg protein) without altering the K(D) (untreated, 7.16; +CsA, 7.06 nM). CsA also significantly increased the level of [Ca(2+)]i measured in cultured mTAL cells both in the basal state (-CSA, 72.2; +CsA, 93.4 nM/10(6) cells) and in the presence of Ang II (-CSA, 97.8; +CsA, 206.3 nM/10(6) cells). CONCLUSIONS: These findings suggest that the increase in Ang II AT(1) receptors and [Ca(2+)]i caused by CsA may be involved in the mechanism(s) responsible for CsA nephrotoxicity. PMID- 12897083 TI - Stimulation of protein degradation by low pH in L6G8C5 skeletal muscle cells is independent of apoptosis but dependent on differentiation state. AB - BACKGROUND: In chronic renal failure, metabolic acidosis increases protein degradation (PD) in skeletal muscle, an effect which in vivo requires glucocorticoid (GC). This disorder is poorly understood, but can be studied in vitro using L6G8C5 rat skeletal muscle cells. Two potential confounding factors in studies of PD in culture are apoptosis and dedifferentiation, both of which resemble catabolic states. The aim of this study was to determine the extent to which these factors contribute to the observed effects of acid and GC on PD. METHODS: PD was measured in intact cells by pre-labelling cell protein with [(14)C]phenylalanine. Apoptosis was assessed morphologically by staining DNA with Hoechst 33342, by terminal deoxynucleotide transferase-mediated nick-end labelling and by cell-surface binding of Annexin V. Differentiation was assessed morphologically from myotube fusion and from activity of the marker enzyme creatine phosphokinase (CPK). RESULTS: In undifferentiated myoblasts, pH had no detectable effect on apoptosis provided that serum was present and GC (dexamethasone; 5 nmol/l) decreased apoptosis. In spontaneously fused cultures in 2% serum, inhibition of apoptosis with caspase-3 inhibitor (C3I; Ac-Asp-Met-Gln Asp-CHO; 50 micro mol/l) only decreased PD by 9% at pH 7.4. In contrast, the proteasome inhibitor MG132 decreased PD by 79%. Acid (pH 7.1) increased PD, with no requirement for GC, and this effect was blocked by MG132, but not by C3I. Differentiation was unaffected by 1-4 days of exposure to acid or GC. However, differentiation to myotubes led to decreased sensitivity of PD to acid. This effect of acid was lost completely in highly fused myotubes, but was partly restored by 500 nmol/l dexamethasone. CONCLUSIONS: Stimulation of PD in these cells by acid and GC is not an artefact of apoptosis or dedifferentiation, but differentiation state does determine whether PD responds spontaneously to acid or (as in vivo) only does so in the presence of GC. PMID- 12897084 TI - Glucocorticoid antagonist RU38486 fails to block acid-induced muscle wasting in vivo or in vitro. AB - BACKGROUND: Increased protein degradation during metabolic acidosis contributes to muscle wasting in uraemia. Adrenalectomy experiments in severely acidotic rats (arterial pH approximately 7.15) have shown that this is prevented in the absence of glucocorticoid. It should therefore be possible to block such muscle wasting with glucocorticoid receptor antagonist 11beta-(4-dimethylaminophenyl)-17beta hydroxy,-17a-(prop-1-ynyl)-estra-4,9-dien-3-one (RU38486). METHODS: The effect of oral RU38486 (50 mg/kg body weight/day) was studied in vivo by administration to rats receiving dietary HCl supplements which yielded moderate acidosis (plasma HCO(3)(-) 19.7 +/- 1.2 mmol/l), comparable with that observed in uraemia. The effect of the glucocorticoid dexamethasone (DEX) (up to 500 nmol/l) and RU38486 (up to 5 micro mol/l) was also studied in vitro in acidified cultures of L6-G8C5 rat skeletal muscle cells. RESULTS: In vivo 15 days of moderate acidosis slowed weight gain and induced muscle wasting (6% weight loss in gastrocnemius with a commensurate decline in muscle protein) but, at this level of acidosis, muscle protein degradation showed no detectable increase. Wasting was not inhibited by RU38486 in spite of blockade of 80% of the glucocorticoid receptors in gastrocnemius. Unexpectedly, weight gain was significantly slower in acidotic rats receiving RU38486 than in acidotic rats receiving vehicle. In vitro acid spontaneously stimulated protein degradation, but even under strongly acidic conditions (pH 7.1) this was only weakly and transiently stimulated by 5 nmol/l DEX and transiently blunted by 5 micro mol/l RU38486. In contrast, as little as 1 nmol/l insulin-like growth factor I (IGF-I) almost abolished the effect of acid and this was partly restored by 5 nmol/l DEX. CONCLUSIONS: IGF-I is a potent determinant of acid-induced protein degradation in vitro and is antagonized by glucocorticoid. If glucocorticoid acts in this indirect way in vivo this may explain why, in moderate metabolic acidosis with intact adrenal glands, the action of RU38486 via glucocorticoid is too weak to be of therapeutic value. PMID- 12897085 TI - Hypoalbuminaemia enhances the renal vasoconstrictor effect of lysophosphatidylcholine. AB - BACKGROUND: Lysophosphatidylcholine (LPC) causes vascular dysfunction in vitro. Lipoprotein LPC is increased in hypoalbuminaemia. Albumin binds LPC and restores LPC-induced abnormalities. We hypothesized that in vivo LPC impairs blood flow more in hypoalbuminaemia than in normoalbuminaemia. METHODS: Increasing concentrations of LPC were infused intra-renally in Nagase analbuminaemic rats (NAR) and Sprague-Dawley rats (controls). RESULTS: Intra-renal LPC (0.1 micromol/min, 20 min) reduced renal blood flow (RBF) more (P < 0.01) in NAR (from 8.3 +/- 0.3 to 4.0 +/- 1.1) than in controls (from 7.7 +/- 0.7 to 5.8 +/- 0.5 ml/min/g kidney). Lysophosphatidylethanolamine had no effect. After stopping LPC, RBF recovery was delayed in NAR [median 90 (range: 70-90) vs 45 min (40-60), P < 0.01]. Intravenous bovine serum albumin (BSA) prevented LPC-induced vasoconstriction in both strains. Prolonging LPC for 60 min delayed recovery of RBF. In this setting, intra-renal BSA completely restored RBF in 75 min (30-90), while intra-renal saline over 75 min only resulted in 33 +/- 13% recovery (P < 0.01). Baseline renal LPC content was unchanged in NAR. However, intra-renal LPC infusion doubled renal LPC content in NAR, but had no effect in controls. CONCLUSIONS: In NAR, baseline RBF and renal LPC content are normal. However, exposure of NAR to LPC results in much more vasoconstriction and accumulation of LPC than in normoalbuminaemia. Addition of albumin prevents and restores LPC induced vasoconstriction. PMID- 12897086 TI - In situ activation pattern of Met docking site following renal injury and hypertrophy. AB - BACKGROUND: Hepatocyte growth factor/scatter factor (HGF/SF) binds to its tyrosine kinase receptor, Met, thereby stimulating diverse cellular responses. The multifunctional docking site in the C-terminal domain mediates the signal of phosphorylated Met receptors to multiple transducers. The tyrosine at position 1356 of the Met docking site is crucial for cell motility and morphogenesis. METHODS: We examined the in situ distribution patterns of the Tyr1356 phosphorylated form of Met with a novel monoclonal antibody following renal injury and renal hypertrophy in rats. Sections of the kidney following either sham operation, transient ischaemia of one kidney or unilateral nephrectomy were analysed using indirect immunofluorescence staining and confocal laser scanning microscopy analysis of total Met protein levels and Tyr1356-phosphorylated Met (Met and pMet, respectively). RESULTS: At 6 h post-treatment, pMet increases in ischaemic kidneys compared with sham-operated kidneys, and these changes become substantial after 48 h in both medulla and cortex of ischaemic kidneys (P < 0.001). We also show significant up-regulation of Met predominantly in the medulla of ischaemic kidneys, 48 h following injury (P < 0.009). Inter-estingly, the stimulus for hypertrophy in the remnant kidney after uninephrectomy and the contra-lateral kidney during ischaemia is not accom-panied by significant up regulation of Met or pMet staining compared with sham operation at both time points. CONCLUSIONS: We demonstrate in this work, for the first time, in situ detection of tyrosine kinase growth factor receptor docking site activation during pathological processes in the kidney. Using this methodology, we show a significant increase in Met docking site activity in both renal medulla and cortex solely following stimulation by ischaemia and repair. PMID- 12897087 TI - Up-regulation of organic anion transporter 1 protein is induced by chronic furosemide or hydrochlorothiazide infusion in rat kidney. AB - BACKGROUND: Thiazide and loop diuretics are secreted from the proximal tubule via the organic anion transport system to reach their principal sites of action. Recently, a multispecific organic anion transporter 1 (OAT1) was identified in rat kidney and was localized to the basolateral membrane of the S2 segment in the proximal tubule. We postulated that interactions between thiazide or loop diuretics and OAT1 may play a role in the adaptation to long-term diuretic use, and investigated whether OAT1 is regulated in vivo by chronic administration of diuretics at the protein level. METHODS: Semi-quantitative immunoblotting and immunohistochemistry were carried out in kidneys from male Sprague-Dawley rats using a polyclonal peptide-derived antibody to OAT1. Furosemide (12 mg/day/rat, n = 6), hydrochlorothiazide (3.75 mg/day/rat, n = 6) or vehicle (1.7% ethanolamine, n = 6) were infused subcutaneously for 7 days using osmotic minipumps. Experimental and vehicle-control rats were pair-fed, and two bottles of drinking water were provided, one containing tap water and the other containing a solution of 0.8% NaCl with 0.1% KCl. RESULTS: Overt diuretic responses were observed to both furosemide and hydrochlorothiazide infusions. There were no differences in body weight or creatinine clearance between the experimental and control rats. Although OAT1 protein abundance in cortical homogenates was increased by furosemide infusion (271 +/- 35 vs 100 +/- 15%, P < 0.05), Na-K-ATPase alpha1 subunit protein abundance was not affected (113 +/- 14 vs 100 +/- 8%, P = 0.42). Immunohistochemical localization in tissue sections confirmed a strong increase in OAT1 expression in the basolateral membrane of the S2 segment of proximal tubule. OAT1 protein abundance in cortical homogenates was also increased by hydrochlorothiazide infusion (181 +/- 25 vs 100 +/- 7%, P < 0.01), whereas Na-K ATPase alpha1 subunit protein abundance was not affected (105 +/- 4 vs 100 +/- 4%, P = 0.34). CONCLUSION: Chronic furosemide or hydrochlorothiazide infusion caused increases in OAT1 protein abundance in rat kidney. These results suggest that OAT1 may be up-regulated in vivo by substrate stimulation at the protein level. PMID- 12897088 TI - Vitamin C augments the renal response to L-arginine in smokers. AB - BACKGROUND: In the coronary and the forearm circulations, endothelium-dependent vasomotion is impaired in smokers, but can be augmented by -arginine or vitamin C. We examined whether smoking similarly affects the renal circulation. METHODS: In 20 smokers (age 26 +/- 4 years) and in 20 non-smokers (age 28 +/- 3 years) changes of renal plasma flow (RPF), glomerular filtration rate (GFR), blood pressure and heart rate in response to the subsequent intravenous infusions of N(G)-monomethyl--arginine (L-NMMA), -arginine and -arginine plus vitamin C were studied by use of a constant infusion input clearance technique. RESULTS: Systemic haemodynamic parameters did not differ between smokers and non-smokers during each experimental phase. At baseline, RPF and GFR were similar between the groups. The infusion of L-NMMA led to a similar decrease of RPF, while GFR did not change in either group. During the infusion of -arginine RPF increased similarly. Finally, the co-infusion of -arginine plus vitamin C led to a significantly greater increase of RPF (+277 +/- 395 vs +79 +/- 76 ml/min, P = 0.03) and GFR (+12.1 +/- 10.6 vs +3.4 +/- 11.2 ml/min, P = 0.02) in smokers as compared to non-smokers. CONCLUSIONS: L-NMMA-induced vasoconstriction of the renal vasculature was similar in smokers compared to non-smokers. -arginine alone induced a similar increase of RPF. The co-infusion of vitamin C and -arginine led to a greater increase of RPF and GFR in smokers. This might suggest that oxidative stress is increased in the renal vasculature of smokers. PMID- 12897089 TI - Oxidative stress-related factors in Bartter's and Gitelman's syndromes: relevance for angiotensin II signalling. AB - BACKGROUND: Bartter's and Gitelman's syndromes (BS/GS) have a blunted Gq protein mediated cell signalling despite high circulating angiotensin II (Ang II) levels. This is associated with reduced Galphaq gene expression, intracellular inositol trisphosphate and Ca(++) release, PKC activity and cell reactivity. Ang II is a powerful stimulator of vascular oxidases but BS/GS patients show reduced total volatile LDL oxidation products and reduced LDL susceptibility to oxidation suggesting low level of oxidative stress. Therefore, we evaluated oxidative stress-related proteins in plasma and monocytes of patients with BS/GS, at baseline and after Ang II stimulation. METHODS: In two BS and seven GS patients, biochemically and genetically characterized, and in 10 age- and sex-matched control subjects, we measured total plasma antioxidant power (AOP), plasma peroxynitrite level and gene expression of the NADH/NADPH oxidase subunit p22(phox), TGFbeta and haeme oxygenase-1 (HO-1) in circulating monocytes in basal condition and after stimulation with Ang II. Furthermore, we investigated the C(242)T polymorphism of p22(phox), whose topography in a potential haeme-binding site suggests a role in the regulation of oxidative stress. RESULTS: AOP was higher in BS/GS patients than in controls (3.27 +/- 0.95 mmol/l vs 1.05 +/- 0.16, P = 0.002), together with higher plasma renin activity and aldosterone level (9.88 +/- 4.64 vs 0.95 +/- 0.08 nmol Ang I/h/ml, P < 0.0001; and 0.73 +/- 0.13 vs 0.18 +/- 0.01 nmol/l, P < 0.0001, respectively). The plasma peroxynitrite level was undetectable both in patients and controls. mRNA expression of p22(phox) and TGFbeta was reduced in BS/GS patients compared to controls [0.35 +/- 0.08 vs 0.53+/-0.05 densitometric units (d.u.), P = 0.005, and 0.82 +/- 0.07 vs 1.15 +/- 0.25 d.u., P = 0.006, respectively]. HO-1 mRNA was increased in BS/GS patients in comparison to controls (0.88 +/- 0.07 vs 0.78 +/- 0.11 d.u., P = 0.037). After acute Ang II exposure, p22(phox), TGFbeta and HO-1 gene expression significantly increased only in controls (from 0.59 +/- 0.12 to 0.96 +/- 0.11, P < 0.001, from 0.97 +/- 0.1 to 1.27 +/- 0.22, P < 0.008, and from 0.62 +/- 0.1 to 0.82 +/- 0.09, P < 0.001, respectively). Finally, C(242)T polymorphism of p22(phox) was undetectable. CONCLUSIONS: The intracellular responses to Ang II mediated by reactive oxygen species are reduced in BS/GS patients. This may contribute to their vascular hyporeactivity. PMID- 12897090 TI - Glomerular expression of C-C chemokines in different types of human crescentic glomerulonephritis. AB - BACKGROUND: Crescentic glomerulonephritis (CGN) presents a rapidly progressive glomerulonephritis clinically, in which macrophages play a crucial role in the pathogenesis. However, the precise molecular mechanism of macrophage recruitment and activation has not been fully elucidated. C-C chemokines, monocyte chemoattractant protein-1 (MCP-1) and macrophage inflammatory protein-1alpha and beta (MIP-1alpha and MIP-1beta), are major chemoattractants for macrophages. We attempted to study the expression of C-C chemokines and their correlation with CD68-positive macrophages in crescentic glomeruli to investigate further their possible roles in crescent formation and progression to fibrosis in different types of human CGN. METHODS: The expression of MCP-1, MIP-1alpha, MIP-1beta and CD68 was detected in glomeruli with different forms of crescents (cellular, fibrocellular and fibrous crescents) by immunohistochemistry in serial sections of renal biopsies taken from 32 patients with biopsy-proven CGN including eight patients with anti-glomerular basement membrane (GBM) disease (type I CGN), 12 patients with immune complex-mediated CGN (type II CGN) and another 12 patients with pauci-immune CGN (type III CGN) enrolled in this study. Eight normal human kidneys were obtained from cadaveric renal transplant donors whose kidneys were technically unsuitable for transplantation, serving as controls. RESULTS: MCP-1, MIP-1alpha, MIP-1beta and CD68 were undetectable in glomeruli of normal kidney. In crescentic biopsies, MCP-1, MIP-1alpha, MIP-1beta and CD68 were detected in fibrocellular crescents and were even more prominent in cellular crescents, but were undetectable in fibrous crescents. Using consecutive sections for staining, it was demonstrated that a high proportion of infiltrating CD68-positive macrophages, mainly localized to the area of the expression of chemokines, were MCP-1, MIP-1alpha and MIP-1beta positive in crescents. Chemokines were expressed mainly by CD68-positive macrophages and parietal epithelial cells in crescents. The number of MCP-1- and MIP-1alpha-positive cells in glomeruli with cellular crescents was positively correlated with the number of CD68-positive cells (r = 0.568 and 0.749, respectively, both P < 0.01). The number of MCP-1- and MIP 1alpha-positive cells and the incidence of Bowman's capsule rupture in glomeruli of patients with type I CGN were higher than those of type II and type III CGN. CONCLUSIONS: These observations suggest that the expressed C-C chemokines, MCP-1, MIP-1alpha and MIP-1beta, may mediate the inflammatory process of crescent formation and progression to fibrosis. The strong correlation of MCP-1 and MIP 1alpha with infiltrating macrophages within glomeruli with cellular crescents suggested that these chemokines might be of particular importance for macrophage recruitment to this site. MCP-1 and MIP-1alpha were correlated to type I CGN with its more severe inflammatory course and worse prognosis. The variance of glomerular expression of C-C chemokines may contribute to the difference in histopathological features and prognosis in these three types of CGN. PMID- 12897091 TI - Effects of the C677T and A1298C polymorphisms of the MTHFR gene on the genetic predisposition for diabetic nephropathy. AB - BACKGROUND: Methylenetetrahydrofolate reductase (MTHFR) is a regulatory enzyme of homocysteine metabolism. The C677T polymorphism of the MTHFR gene has been reported to be associated with elevated plasma homocysteine in patients with low folic acid intake. A recently reported second common polymorphism, A1298C, may increase homocysteine, but only in individuals carrying the T677 allele. This study aimed to investigate the influence of the C677T and A1298C polymorphisms of the MTHFR gene on the development of diabetic nephropathy in Caucasian patients with type 2 diabetes. METHODS: We genotyped 429 type 2 diabetic patients for the C677T and A1298C polymorphisms using standard PCR-based protocols, and divided them into three groups based on renal status: 159 patients with normoalbuminuria, 149 with microalbuminuria, and 121 with persistent proteinuria and chronic renal failure (CRF). The C677T and A1298C genotype frequencies were compared among the three groups. RESULTS: Although the frequencies of the CT and TT genotypes of the C677T polymorphism tended to increase with each stage of diabetic nephropathy (53, 56 and 63% in normoalbuminuria, microalbuminuria and proteinuria/CRF, respectively), these differences were not significant. When male and female patients were analysed separately, the effect was seen only in males. The CT + TT genotype was present in 46% of male patients with normoalbuminuria, in 57% with microalbuminuria and in 68% with proteinuria/CRF (OR = 2.46; 95% CI 1.13-5.38). There were no differ-ences in the A1298C polymorphism among the three groups. CONCLUSIONS: These findings indicate that the C677T polymorphism is a risk factor for diabetic nephrop-athy in male patients with type 2 diabetes. PMID- 12897092 TI - A tricontinental view of IgA nephropathy. AB - BACKGROUND: The purpose of this retrospective study was to analyse patients from four centres in three continents to determine if differences in long-term outcome of IgA nephropathy (IgAN) are explained by clinical and laboratory features at presentation. METHODS: The study included 711 adults with biopsy-proven IgAN from Glasgow, UK (n = 112), Helsinki, Finland (n = 204), Sydney, Australia (n = 121) and Toronto, Canada (n = 274). Data collected from time of presentation to a nephrologist were age, gender, 24-h urine protein excretion (UP(0)), mean arterial pressure (MAP(0)) and creatinine clearance (CrCl(0)). Outcomes were slope of creatinine clearance (CrCl) and renal survival. RESULTS: At presentation there was significant vari-ability in baseline clinical features with patients from Helsinki having the lowest median UP(0), lowest MAP(0) and highest CrCl(0), all suggesting milder disease. There was significant variability in renal survival between centres with 10-year actuarial survival of 95.7, 87.0, 63.9 and 61.6% in Helsinki, Sydney, Glasgow and Toronto, respectively (P < 0.0001; log rank). Cox proportional hazards model revealed lower age(0) and lower CrCl(0) were significant independent predictors of reduced renal survival. In addition, patients from Helsinki and Sydney but not Glasgow had significantly longer renal survival than patients from Toronto. Median slope of CrCl varied by region from 1.24 ml/min/year in Helsinki, to -3.99 ml/min/year in Toronto (Kruskal-Wallis H test P < 0.0001). By multivariate analysis older age(0), higher CrCl(0) and lower UP(0) were independently associated with slower progression. Subjects from Helsinki had a significantly slower deterioration independent of the other clinical parameters at presentation. When the 269 patients presenting with CrCl(0) <75 ml/min were analysed separately there was no independent centre effect. CONCLUSIONS: The findings are consistent with the hypothesis that geographical variability in long-term outcome of IgAN is explained by lead-time bias and inclusion of milder cases in centres with apparent good outcome, but do not exclude the possibility that some of the variability is due to other factors such as genetics, diet or treatment. PMID- 12897093 TI - Moderate alcohol intake and renal function decline in women: a prospective study. AB - BACKGROUND: The impact of moderate alcohol consumption on renal function has important public health implications given the high prevalence of alcohol use. Experimentally, alcohol may adversely affect renal function, but clinical data are limited and no large, prospective studies have examined this issue. METHODS: In a prospective study of 1658 nurses enrolled in the Nurses' Health Study, we sought to determine if there was an association between moderate alcohol consumption and rate of decline in renal function. Daily alcohol intake was measured in 1990, 1994 and 1998 using a detailed questionnaire. Maximum daily alcohol intake was measured in 1988. Creatinine, measured from blood samples provided in 1989 and 2000, was used to estimate glomerular filtration rate (GFR) and creatinine clearance (CCr). RESULTS: Compared to individuals with no alcohol intake, the odds ratios (ORs) for developing a >or=25% estimated GFR decline were: 0.98 (95% CI: 0.72-1.32) for 0.1-4.9 g/day, 0.83 (95% CI: 0.56-1.21) for 5 14.9 g/day and 0.81 (95% CI: 0.50-1.31) for 15-59.9 g/day. For women with hypertension (n = 726), the ORs for a >or=25% estimated GFR decline were: 0.98 (95% CI: 0.53-1.21) for 0.1-4.9 g/day, 0.62 (95% CI: 0.34-1.12) for 5-14.9 g/day and 0.53 (95% CI: 0.25-1.12) for 15-59.9 g/day. CONCLUSIONS: Moderate alcohol consumption had no substantial adverse effect on renal function in women over an 11 year follow-up period. PMID- 12897094 TI - Renal function and cardiovascular risk markers in a remote Australian Aboriginal community. AB - BACKGROUND: Australian Aborigines living in remote areas have exceedingly high rates of renal failure together with increased cardiovascular morbidity and mortality. To examine the basis of this association, we studied markers of renal function and cardiovascular (CV) risk in a coastal Aboriginal community in a remote area of the Northern Territory of Australia. End-stage renal disease (ESRD) incidence rates in that community are 15 times the national non-Aboriginal rate and CV mortality rates in the region are increased 5-fold. METHODS: A cross sectional community survey was conducted. Markers of early renal disease examined included urine albumin/creatinine ratio (ACR), serum creatinine concentration and calculated glomerular filtration rate (GFR). CV risk markers included blood pressure as well as measures of glycaemia, diabetes and serum lipids. RESULTS: The study group included 237 people, 58% of the adult population of the community. The crude prevalence of microalbuminuria (urine ACR: 3.4-33.9 g/mol, 30-299 mg/g) was 31% and of overt albuminuria (urine ACR: >or=34 g/mol, >or=300 mg/g), 13%. The prevalence of overt albuminuria increased with age, but the prevalence of microalbuminuria was greatest in the 45-54 year age group. Microalbuminuria was associated with increasing body mass index, whereas overt albuminuria was associated with increasing glycated haemoglobin (HbA1c) and systolic blood pressure and a history of diabetes. The prevalence of elevated serum creatinine concentration (>or=120 micromol/l) was 10%. GFR (calculated using the MDRD equation) was <60 ml/min/1.73 m(2) in 12% and 60-79 ml/min/1.73 m(2) in a further 36% of the study population. Although many people with albuminuria had well preserved GFRs, mean GFR was lower in people with higher levels of albuminuria. CONCLUSIONS: The high prevalence of markers of renal disease in this community was consistent with their high rates of ESRD. The distribution of microalbuminuria suggested a 'cohort effect', representing a group who will progress to overt albuminuria. The powerful association of renal disease markers with CV risk factors confirms a strong link between renal and CV disease in the early, asymptomatic stages of each. Thus, pathologic albuminuria, in part, might be a manifestation of the metabolic/haemodynamic syndrome and both conditions might arise out of a common menu of risk factors. Hence, a single agenda of primary and secondary intervention may benefit both. PMID- 12897095 TI - Regional variability in the incidence of end-stage renal disease: an epidemiological approach. AB - BACKGROUND: Regional variability in the incidence of end-stage renal disease (ESRD) in Austria is reported. Our aim was to investigate the reason for low rates in the state of Tyrol. METHODS: ESRD incidence data were obtained from the Austrian Dialysis and Transplantation Registry. Additional sources were two health interview surveys, the Hospital Discharge Registry, the Mortality Registry and the Drug Wholesale Registry. RESULTS: Between 1995 and 1999, 4811 new cases of ESRD were recorded; the state of Tyrol (T) had a mean annual, age-adjusted incidence of 97.9/1 000 000 population [95% confidence interval (CI) 86.9-109.1], a number significantly lower than that for the rest of Austria [(RA), 120.9 (95% CI 116.9-124.5); P < 0.001]. This was due mainly to a difference in the incidence of ESRD patients with type 2 diabetes mellitus [(DM-2) T = 12.2 (95% CI 8.2-16.2) vs RA = 28.9 (95% CI 27.2-30.6); P < 0.001]. When these patients were excluded, the difference in the overall ESRD incidence disappeared. When data from various registries were analysed for the prevalence of DM, a highly significant correlation was found between ESRD incidence and DM. CONCLUSION: We conclude that the variability in the ESRD incidence in Austria is explained mainly by regional differences in DM-2. Data from similar studies might be useful for predictions concerning resource allocation for ESRD programmes in the future. PMID- 12897096 TI - Relationship of non-LDL-bound apo(a), urinary apo(a) fragments and plasma Lp(a) in patients with impaired renal function. AB - BACKGROUND: Plasma lipoprotein (a) [Lp(a)] has been shown to be a risk factor for atherosclerosis in numerous studies. However, the catabolism of this lipoprotein is not very clear. We and others have shown that Lp(a) is excreted into urine in the form of fragments. Lp(a) has also been shown to exist in a low-density non lipoprotein (LDL)-bound form. Since Lp(a) is increased in all forms of kidney disease with reduced excretory kidney function and decreased excretion of apo(a) fragments could be partially responsible for this increase, we investigated the relationship of non-LDL-bound apo(a), urinary apo(a) fragments and plasma Lp(a) in patients with impaired renal function. METHODS: Plasma Lp(a), non-LDL-bound apo(a) and urinary apo(a) fragments were measured in 55 kidney disease patients (28 males and 27 females) and matched controls. RESULTS: Plasma Lp(a) and non-LDL bound apo(a) were increased in patients, whereas urinary apo(a) was decreased, especially in patients with a creatinine clearance < 70 ml/min. There was a significant correlation between plasma Lp(a) and non-LDL-bound apo(a) in patients and controls. CONCLUSION: We conclude that decreased urinary apo(a) excretion could be one possible mechanism of increased plasma Lp(a) and non-LDL-bound apo(a) in patients with decreased kidney function. PMID- 12897097 TI - Interferon and ribavirin treatment in patients with hepatitis C-associated renal disease and renal insufficiency. AB - BACKGROUND: Hepatitis C virus (HCV) infection is associated with renal manifestations, such as membranoproliferative glomerulonephritis (MPGN) with or without cryoglobulinaemia, membranous glomerulonephritis (MGN) and focal segmental glomerulosclerosis (FSGS). Standard treatment for HCV is interferon and ribavirin, but in renal insufficiency ribavirin has been contraindicated due to fear of side effects. METHODS: Seven patients, two with cryoglobulinaemia, vasculitic manifestations and glomerulonephritis (GN), four with MPGN and one with FSGS were treated with a combination of interferon and ribavirin. Two patients were given pegylated interferon and ribavirin. All patients had at presentation renal insufficiency, with a glomerular filtration rate (GFR) between 10 and 65 ml/min. One patient had HCV genotype 1, the remainder 2 and 3. Duration of therapy was according to genotype (6-12 months). Ribavirin in plasma was monitored by high-performance liquid chromatography (HPLC) to avoid over-dosing, aiming at a target concentration of 10-15 micromol/l. The main side effect of ribavirin, haemolytic anaemia, was monitored closely with haemoglobin controls. RESULTS: Six of seven patients became HCV-RNA-PCR negative and four of seven have maintained both virological and renal remission. One of seven has maintained virological and partial renal remission. One patient did not tolerate interferon, but is in renal remission with low-dose ribavirin. One vasculitis patient responded with complete remission, but relapsed virologically and had a minor vasculitic flare after 9 months. Only one patient with vasculitis had low-dose immunosuppression in addition to anti-viral therapy. Average daily ribavirin dose was 200-800 mg. Ribavirin-induced anaemia was managed in five of seven patients with low-dose iron and erythropoietin between 4000 and 20 000 IU/week. CONCLUSIONS: Interferon and ribavirin can with reasonable safety be used in HCV related vasculitis and GN irrespective of renal function. PMID- 12897098 TI - alpha-Galactosidase A deficiency in Dutch patients on dialysis: a critical appraisal of screening for Fabry disease. AB - INTRODUCTION: Fabry disease or alpha-galactosidase A (alpha-Gal A) deficiency is an X-linked lysosomal storage disorder that often leads to renal insufficiency in males and occasionally in females. The disease is rare, but its prevalence may be underestimated due to its variable clinical picture. Enzyme supplementation therapy with rHu-alphaGal A is currently available. Limited experience has so far shown that therapy may at best stabilize renal function. Despite these preliminary findings, much effort is being put into screening high-risk groups for undiagnosed alpha-Gal A deficiency. We studied the prevalence of alpha-Gal A deficiency in a Dutch dialysis cohort to establish possible underdiagnosis. We discuss the benefits of screening for Fabry disease. METHODS: Activity of alpha Gal A in whole blood was measured in a group of 508 male Dutch dialysis patients. RESULTS: Of the 508 patients studied only one patient, already known with Fabry disease, had a alpha-Gal A deficiency, a prevalence of 0.22% (95 CI 0-1.1%). CONCLUSIONS: No undiagnosed Fabry patients were found, indicating that in our studied cohort there is no large-scale underestimation of its prevalence. Even though screening of dialysis patients for Fabry disease might identify patients who remain otherwise unrecognized, screening of high-risk populations for alpha Gal A deficiency should be carried out with caution since long-term efficacy of treatment is currently unknown. PMID- 12897099 TI - Interactions between dialysis-related volume exposures, nutritional surrogates and mortality among ESRD patients. AB - BACKGROUND: Interdialytic weight gain is used as a surrogate for volume expansion in haemodialysis patients and as an indicator of non-compliance. Increased weight gain is associated with both a greater mortality risk and better nutrition indices. This analysis characterizes the association between dialysis-related volume expansion and mortality in the context of its interaction with nutritional surrogates. METHODS: All patients receiving haemodialysis through Fresenius Medical Care-North America during 1998 were included. The percentage reduction in weight or intradialytic weight loss (IDWL%) was defined as the difference between the average of pre- and post-dialysis weights from the last 3 months of 1997 expressed as a percentage of post-dialysis weight. Associations between IDWL% and clinical and demographic variables were estimated using linear regression. The association between mortality risk and IDWL% was estimated using Cox proportional hazards regression. RESULTS: Younger age, male gender, the presence of diabetes mellitus, decreasing cholesterol, post-dialysis weight and pre-dialysis blood pressure (systolic and pulse pressure) were associated with increased IDWL%. Increasing IDWL% was associated with increasing phosphorus, creatinine, albumin, potassium and urea reduction ratio. Increasing IDWL% was significantly associated with mortality at 1 year [hazard ratio (HR) = 1.07, P = 0.003]. Among patients with diabetes mellitus, increasing IDWL% was associated with a mortality HR of 1.03 (P = 0.02). Among patients without diabetes mellitus, increasing IDWL% was not associated with an increased mortality risk. Increasing IDWL% is associated with a greater mortality risk among patients with creatinine <7.26, which failed to remain significant for patients whose creatinine was >or=7.26 mg/dl. Increasing IDWL% is associated with a greater mortality risk among patients with greater post-dialysis weight, greater body mass index and lower serum sodium measurements. CONCLUSIONS: This study confirms and extends the findings of the deleterious association between increasing IDWL% and mortality among patients with diabetes mellitus and among subgroups based on serum creatinine and body weight. The putative deleterious effect of dialysis-related volume expansion on mortality must be interpreted in the context of the patient's diabetic and nutritional status. PMID- 12897100 TI - Mononuclear leukocyte apoptosis in haemodialysis patients: the role of cell thiols and vitamin E. AB - BACKGROUND: An increased apoptotic rate of peripheral blood mononuclear leukocytes (PBMLs) in haemodialysis (HD) patients has been reported in several studies, but its underlying mechanisms remain poorly understood. Oxidant stress is a well known cause of cell damage, and several lines of evidence suggest that it might influence the induction and signalling steps of mononuclear cell apoptosis through different mechanisms so as to provoke disturbances of the intracellular pool of thiols (SHi). In this study, we investigated the in vitro apoptotic rate and SHi of PBMLs in end-stage renal disease (ESRD) patients on HD or peritoneal dialysis (PD). METHODS: Apoptosis and SHi were evaluated in vitro in PBMLs obtained from 40 ESRD patients (HD, n = 30 and PD, n = 10) and 10 healthy controls. A subgroup of HD patients was also studied before and after 1 month of treatment with a vitamin E-coated dialyser (CL-E). Cell thiols and viability were also assessed in the monocyte-like cell line U937 and PBMLs after incubation in the presence of uraemic plasma with or without supplementation of the antioxidants vitamin E (70 micro M) or N-acetyl-cysteine (NAC) (0.5 mM). RESULTS: After 24 h in culture, the PBMLs of HD patients, but not those of CAPD patients, showed an apoptotic rate twice that of healthy controls and a 40% decrease of SHi levels (P < 0.01 in both). A negative correlation between the apoptotic rate and SHi was observed in both patients and controls (r = 0.648, P < 0.001). Plasma and ultrafiltrate samples from HD patients contained solutes (mainly in the low-middle molecular weight range) able to trigger apoptosis and oxidative stress in U937 cells. The treatment of HD patients with CL-E, as well as the in vitro supplementation of U937 cells with vitamin E or NAC during the exposure to uraemic plasma, decreased the rate of apoptosis and partially restored SHi. CONCLUSIONS: This study showed an association between an increased apoptotic rate and decreased SHi in PBML of HD patients, but not of CAPD patients. These changes are partially due to different pro-apoptogens that accumulate in the plasma and are at least partially prevented by exogenous antioxidants able to restore SHi, such as vitamin E or thiol suppliers. PMID- 12897101 TI - Modification of elastin by pentosidine is associated with the calcification of aortic media in patients with end-stage renal disease. AB - BACKGROUND: Calcification of the media of arteries is common in patients with end stage renal disease (ESRD) undergoing haemodialysis and is a major cause of arteriosclerosis. The aim of this study was to clarify the role of glycoxidative modification of elastin in the calcification of aortic media in this group of patients. METHODS: Samples of tunica media were obtained from non-atherosclerotic areas of the aortas of cadavers of seven non-diabetic patients with ESRD (age 65.5 +/- 10.6 years) and 10 age-matched controls (age 61.1 +/- 10.3 years). The localization of pentosidine, a major glycoxidation product, and calcium deposits in the media were examined using immunohistochemical and von Kossa staining, followed by orcein staining for elastin fibres. Tissue levels of pentosidine and calcium were measured in elastase-digested media using reversed high-performance liquid chromatography and atomic absorption spectrophotometry, respectively. RESULTS: In aortic media, but not intima, immunostained pentosidine was observed along elastin fibres or in the extracellular spaces between them. Early calcification was manifest as small punctate calcified deposits along elastin fibres in the media. Advanced calcification was found as large, confluent calcified deposits in extracellular spaces between elastin fibres. Double staining showed co-localization of pentosidine and calcified deposits in the media. Both the staining density of pentosidine and calcification were more prominent in ESRD patients than in controls. The mean medial contents of both elastin-associated pentosidine and calcium were significantly higher in ESRD patients than in controls. In ESRD patients, the level of calcium in elastase digested media correlated significantly with pentosidine levels, which increased in parallel with the duration of haemodialysis. CONCLUSIONS: Our results indicate that glycoxidative modification of elastin in aortic media may be involved in the enhancement of medial calcification in ESRD patients on haemodialysis. PMID- 12897102 TI - Serum troponin T measurement in patients with chronic renal impairment predicts survival and vascular disease: a 2 year prospective study. AB - BACKGROUND: Cardiovascular mortality in end-stage renal failure patients is high and early risk stratification in these patients may aid clinical management improving outcomes. Cardiac troponin T (cTnT) is a component of the cardiac myocyte which is released into the circulation following myocardial necrosis. It has been shown to be of prognostic significance in patients with unstable angina. The role of cTnT in patients with renal disease remains unclear. The aim of this investigation, therefore, was to assess the prognostic significance of cTnT in chronic renal impairment patients, pre-dialysis. METHODS: Ninety-six patients with chronic renal impairment were followed prospectively after cTnT determination by a quantitative laboratory method. The clinical outcomes after 2 years were determined. The measured cTnT values were correlated with biochemical parameters and clinical end-points. RESULTS: A cut-off of 0.1 ng/ml was used in assessing the prognostic significance of cTnT. Twenty-five patients had a cTnT >0.1 ng/ml, whilst 71 had a cTnT 0.1 ng/ml was 42% compared with 14% in those with levels below the cut-off. Thirty-three patients died or had a vascular event. The rate of death or a vascular event in the elevated group was 64% compared with 24% in those with levels below the cut-off. At the end of the study, 23 patients were treated by continuous ambulatory peritoneal dialysis, 29 by haemodialysis, 22 had functioning renal transplants and one patient was not on renal replacement therapy. Factors that were found to significantly affect cTnT were diabetes, age and urea. cTnT was found to be a significant predictor of survival in these patients. Patients with high cTnT values were more likely to end up on haemodialysis. No relation of renal function to cTnT level was found. CONCLUSIONS: These results show that in patients with renal impairment, the measurement of cTnT prior to commencing renal replacement is a significant independent predictor of survival. cTnT did show potential as a prognostic test to stratify patients with a high cardiovascular risk and may enable intensive risk factor modification in this patient group. This may be of further use in selection of patients' suitability for renal transplantation. PMID- 12897103 TI - Reduction of hypotensive side effects during online-haemodiafiltration and low temperature haemodialysis. AB - BACKGROUND: This study compares the effect of online-haemodiafiltration (o-HDF, post-dilution mode) with conventional haemodialysis (HD) and 'temperature controlled' HD (Temp-HD) on the haemodynamic stability of hypotension-prone patients. METHODS: Seventeen patients with a history of frequent hypotensive episodes during dialysis sessions were studied, each patient serving as his or her own control. The first 25 HD treatments in comparison with 25 o-HDF sessions were evaluated using identical dialysate temperature. In the second part of the study, o-HDF (n = 25) was compared with Temp-HD (n = 25). In the latter method, the temperature of the dialysate was adjusted to result in identical energy transfer rates to those in the corresponding o-HDF. The number of hypotensive episodes, blood temperature and blood volume regulation were assessed. RESULTS: Symptomatic hypotension was much more frequent during HD (40%) than during o-HDF (4%) (P < 0.001). During o-HDF, an enhanced energy loss within the extracorporeal system occurred (o-HDF, 16.6 +/- 4.0 W; HD, 5.4 +/- 5.1 W; P < 0.0001), despite identical temperature settings for dialysate and substitution fluid. As a result, the blood returning to the patient was cooler during o-HDF than during HD (o-HDF 35 +/- 0.2 degrees C vs HD 36.5 +/- 0.3 degrees C; P < 0.0001). In o-HDF, even in the patients' circulation, the mean blood temperature was lower (o-HDF 36.7 +/- 0.2 degrees C vs HD 36.9 +/- 0.3 degrees C; P < 0.0001) and blood volume was significantly more reduced (o-HDF, 91.8 +/- 3.1%; HD, 94.0 +/- 3.2%; P < 0.05). Energy transfer rates and blood temperature did not differ significantly between o-HDF and Temp-HD. The rate of hypotensive episodes was low and not different between o-HDF (4%) and Temp-HD (4%). Neither was there any significant difference in blood volume reduction. CONCLUSIONS: O-HDF showed a significant reduction of hypotensive episodes compared with HD. Surprisingly, o-HDF resulted in cooling of the blood via enhanced thermal energy losses within the extracorporeal system, despite use of replacement fluid prepared from pre-warmed dialysate. The incidence of symptomatic hypotension was reduced to that of o-HDF by using cooler Temp-HD. Thus, unexpected blood cooling appears to be the main blood pressure stabilizing factor in o-HDF. PMID- 12897104 TI - Effects of argatroban as an anticoagulant for haemodialysis in patients with antithrombin III deficiency. AB - BACKGROUND: In congenital or acquired antithrombin III-deficient patients undergoing haemodialysis, coagulation or residual blood in the blood circuit and dialyser is commonly observed under anticoagulation with heparin. Argatroban, a synthetic thrombin antagonist, directly inhibits thrombin activity in a manner that is different from that of heparin, thereby displaying an anticoagulating effect without the activation of antithrombin III. For this reason, the anticoagulating effect of argatroban in haemodialysis patients with antithrombin III deficiency was investigated. METHODS: A retrospective nationwide survey was conducted among patients with congenital or acquired antithrombin III deficiency who had undergone haemodialysis with argatroban as an anticoagulant from April 1996 to April 2000. Inclusion criteria were patients with antithrombin III activity <70% of normal, and patients in whom blood coagulation or residual blood in the extracorporeal circuit could not be prevented by the use of heparin during haemodialysis. RESULTS: Of 80 patients who underwent haemodialysis with argatroban, 59 met the inclusion criteria. Compared with the data before the administration of argatroban, significant improvements of residual blood in the dialyser and arterial and venous drip chambers were observed at the last administration of argatroban. A significant rise in antithrombin III activity was also observed. Among 80 safety analysis cases, no adverse events were reported in 66 patients (82.5%). As severe adverse events, one showed bleeding tendency and one had prolongation of prothrombin time. CONCLUSION: Argatroban was an effective and safe anticoagulant for haemodialysis in patients with congenital or acquired antithrombin III deficiency. PMID- 12897105 TI - Repeated femoral vein puncturing for maintenance haemodialysis vascular access. AB - BACKGROUND: When access cannot be achieved using a native arteriovenous fistula or a synthetic prosthetic graft, central venous catheters are usually placed. This mode of access is short-lived, prone to infection, stenosis and thrombosis of central veins. To overcome access problems, we developed a new native vascular access ('femoral vein access') and devices. We report here on our experience with the availability, longevity, procedure and morbidity of haemodialysis (HD) using femoral vein access. METHODS: Repeated (three times a week) patient's native femoral vein puncturing has been used as the vascular access (femoral vein access) for maintenance HD in 30 patients (mean age +/- SD: 61.70 +/- 15.27 years old; 18 female/12 male). The femoral vein was punctured beneath the inguinal ligament (on a length ranging from 30 to 100 mm) after disinfection and local anaesthesia. Long (effective length 56 mm) 19- and 18-gauge needles with four side holes were used for the femoral vein puncture as an arterial site of the extracorporeal circuit of HD and shorter (effective length 40 mm) similar gauge needles for the subcutaneous vein puncture used as the return site. The needle is inserted blind into the femoral vein after the femoral artery has been located by palpation and the perception of a pulse. Patients returned home the same day. RESULTS: The mean duration of HD treatment using femoral vein repeated puncture was 4.99 +/- 3.42 years (up to 16.0 years). This represented a total experience of 23 369 femoral vein punctures. The mean blood flow achieved on dialysis was 165 +/- 20 ml/min. The average Kt/V was 1.74 +/- 0.48 per session. CONCLUSIONS: The femoral vein repeated puncture technique has substantial advantages over venous catheters. It does not require surgery, while permitting adequate blood flow. This method can be used as a long-term (over 10 years) blood access. Apart from a few local haematomas, no serious complications have been observed. Moreover, it does not carry a heavy financial burden. PMID- 12897106 TI - No rise in renal Doppler resistance indices at peak serum levels of cyclosporin A in stable kidney transplant patients. AB - BACKGROUND: The measurement of intrarenal resistance indices by duplex ultrasound plays an important role in the follow-up care of renal transplant patients. Increasing resistance indices indicate rejection episodes, but may also occur e.g. in parenchymal renal diseases. As calcineurin inhibitors induce vasoconstriction both in vivo and in vitro, we studied whether peak serum levels of cyclosporin A led to an acute rise in renal resistance indices via the induction of intrarenal vasoconstriction. METHODS: The acute impact of peak serum levels of cyclosporin A on intrarenal resistance indices was studied in 36 patients after allogeneic renal transplantation. All patients were transplanted for > 6 months and received an immunosuppressive treatment comprising cyclosporin A. Intrarenal resistance indices were measured by duplex ultrasound immediately before (trough serum level) and 2 h after (peak serum level) the oral intake of cyclosporin A at the individual maintenance dose. RESULTS: Compared with renal resistance indices measured at trough serum levels [resistive index (RI) 0.72 +/- 0.07; pulsatility index (PI) 1.40 +/- 0.27], values remained unchanged at peak serum levels of cyclosporin A (RI 0.72 +/- 0.08; PI 1.43 +/- 0.31). Renal resistance indices correlated with the age of the patients, but not with mean arterial pressure or time since transplantation. CONCLUSIONS: The oral intake of cyclosporin A does not induce an acute rise in intrarenal resistance indices in stable transplanted patients. Thus, timing of duplex ultrasound examinations with regard to the intake of cyclosporin A is not necessary in these patients. PMID- 12897107 TI - Haemodialysis patients' views on their resuscitation status. AB - BACKGROUND: Guidelines on cardiopulmonary resuscitation (CPR) recommend that decisions about resuscitation are part of every patient's care plan. We aimed to ascertain the initial views of a haemodialysis cohort in the UK regarding their CPR status in the event of an in-hospital cardiac arrest not related to dialysis. METHODS: During outpatient clinic follow-up between January and March 2001, 66 haemodialysis patients were interviewed by one of two interviewers. A standard interview format was used, which included information on complications and outcome of CPR in haemodialysis patients. Patients had no prior opportunity to discuss the issue with a third party. RESULTS: Fifty patients (76%) wished to receive CPR. Five patients decided against CPR, of whom two were depressed. Six patients were undecided. Five patients were excluded from the analysis because of lack of mental capacity or language difficulties. Patients who wished to receive CPR were significantly younger (59 +/- 16 vs 74 +/- 10 years, respectively; P < 0.01) and had a significantly higher serum albumin level compared with those who did not opt for CPR or were undecided. There was no difference in gender, comorbidity, length of time on dialysis, proportion of patients with adequate dialysis and mean haemoglobin level between those who wished CPR and those who did not. CONCLUSIONS: The majority of haemodialysis patients wished to receive CPR. These patients were younger than those who did not opt for CPR. Clearer strategies on third-party consultation and follow-up interviews are needed to guarantee that patients' wishes are respected at all times. PMID- 12897108 TI - Specific characteristics of peritoneal leucocyte populations during sterile peritonitis associated with icodextrin CAPD fluids. AB - BACKGROUND: Icodextrin dialysate used for peritoneal dialysis contains an iso molar glucose polymer solution, which provides sustained ultrafiltration over long dwell times and is considered a valuable approach to reduce intraperitoneal glucose exposure. However, several side effects have been described, including abdominal pain and allergic and hypersensitivity reactions. Also, reactions compatible with chemical peritonitis have been reported. Over the period of a few months (January 2002-May 2002), a remarkable increase in the number of continuous ambulatory peritoneal dialysis (CAPD) patients using icodextrin dialysate diagnosed with sterile peritonitis was observed in our unit. METHODS: Five of the CAPD patients using icodextrin dialysate in our unit and diagnosed with sterile peritonitis were screened for leucocyte count and leucocyte differentiation during a follow-up period of 77 +/- 23 days. In addition, expression of CD14, a receptor for lipopolysaccharide (LPS), on the peripheral and peritoneal monocyte population was analysed. These results were compared to CAPD patients suffering from bacterial peritonitis. RESULTS: The peritoneal leucocyte count of CAPD patients using icodextrin dialysate and diagnosed with sterile peritonitis did not decrease significantly before treatment with icodextrin dialysate was interrupted, whereas it currently disappeared within 2-4 days in proven bacterial peritonitis. The sterile, cloudy icodextrin effluent contained an excess of macrophages on the day of diagnosis, whereas in bacterial peritonitis essentially an increase in the granulocyte population was observed. No elevation in the eosinophil population was observed. In contrast to bacterial peritonitis, we observed no increase in CD14 expression on the peripheral and peritoneal macrophages on the day of presentation and during the follow-up period. CONCLUSIONS: Specific batches of the icodextrin CAPD fluids contain a macrophage chemotactic agent, which causes a sustained inflammatory state in the peritoneal cavity. Because no increase in the expression of the LPS receptor CD14 could be observed, the increased peritoneal leucocyte count is probably not caused by LPS or LPS-like (possibly peptidoglycan-like) contamination. PMID- 12897109 TI - Efficacy and safety of lowering immunosuppression to treat CMV infection in renal transplant recipients on valaciclovir prophylaxis: a pilot study. AB - BACKGROUND: Routine cytomegalovirus (CMV)-pp65 antigenaemia monitoring shows that some patients will develop pp65 antigenaemia during valaciclovir prophylaxis or after cessation of treatment. The aim of this pilot study was to evaluate the safety and efficacy of lowering immunosuppression in kidney transplant recipients who exhibit mildly symptomatic CMV infections while on valaciclovir prophylaxis. METHODS: We selected 12 patients who experienced mildly symptomatic CMV infections defined as a positive CMV-pp65 antigenaemia test associated with either neutropenia, asthenia or arthralgia, but no fever. All of them received prophylaxis with valaciclovir for at least 3 months. Testing for CMV-pp65 antigenaemia was performed weekly for 6 months. RESULTS: The mildly symptomatic infections occurred at a median interval of 69 days after transplantation-during prophylaxis in eight cases and after valaciclovir discontinuation in the other four cases. All of them were effectively managed by lowering immunosuppressive therapy, leading to the disappearance of symptoms and CMV antigenaemia reduction. No immunological complication or recurrence of CMV infection or disease was noted. I.v. ganciclovir never became necessary. CONCLUSION: The mildly symptomatic CMV infections occurring in valaciclovir-treated patients may be managed efficiently and without immunologic complication by lowering immunosuppressive therapy. PMID- 12897110 TI - An unusual case of pseudohyperkalaemia. PMID- 12897111 TI - Amoxicillin-induced acute renal failure. PMID- 12897112 TI - Veno-occlusive disease of the liver in renal transplant patients. PMID- 12897113 TI - An unusual case of unresponsiveness post-renal transplant. PMID- 12897114 TI - A young patient with end-stage renal disease, dyspnoea, weakness, peripheral neuropathy and an unsuspected underlying disease. PMID- 12897115 TI - Ear and kidney malformations with renal failure in an infant: what is the link? PMID- 12897116 TI - The deceiving image: asymptomatic renal malakoplakia in a patient with chronic renal failure. PMID- 12897117 TI - Is coating of tubing required when endothelin-1 is infused intravenously? PMID- 12897118 TI - Cardiac tamponade: an unusual complication of Plasmodium falciparum-induced acute renal failure. PMID- 12897119 TI - Acute renal failure and oligohydramnios induced by magnesium dypirone (metamizol) in a pregnant woman. PMID- 12897120 TI - Striking increase in circulating hepatocyte growth factor during enoxaparin anticoagulated haemodialysis. PMID- 12897121 TI - Tenckhoff catheters: the pull technique. PMID- 12897124 TI - Epigenetic properties and identification of an imprint mark in the Nesp-Gnasxl domain of the mouse Gnas imprinted locus. AB - The Gnas locus in the mouse is imprinted with a complex arrangement of alternative transcripts defined by promoters with different patterns of monoallelic expression. The Gnas transcript is subject to tissue-specific imprinted expression, Nesp is expressed only from the maternal allele, and Gnasxl is expressed only from the paternal allele. The mechanisms controlling these expression patterns are not known. To identify potential imprinting regulatory regions, particularly for the reciprocally expressed Nesp and Gnasxl promoters, we examined epigenetic properties of the locus in gametes, embryonic stem cells, and fetal and adult tissues. The Nesp and Gnasxl promoter regions are contained in extensive CpG islands with methylation of the paternal allele at Nesp and the maternal allele at Gnasxl. Parental allele-specific DNase I-hypersensitive sites were found at these regions, which correlate with hypomethylation rather than actual expression status. A germ line methylation mark was identified covering the promoters for Gnasxl and the antisense transcript Nespas. Prominent DNase I hypersensitive sites present on paternal alleles in embryonic stem cells are contained within this mark. This is the second gametic mark identified at Gnas and suggests that the Nesp and Gnasxl promoters are under separate control from the Gnas promoter. We propose models to account for the regulation of imprinting at the locus. PMID- 12897125 TI - Alternative splicing controls myotonic dystrophy protein kinase structure, enzymatic activity, and subcellular localization. AB - Transcripts of the myotonic dystrophy protein kinase (DMPK) gene, a member of the Rho kinase family, are subject to cell-type specific alternative splicing. An imbalance in the splice isoform profile of DMPK may play a role in the pathogenesis of DM1, a severe multisystemic disorder. Here, we report how structural subdomains determine biochemical properties and subcellular distribution of DMPK isoforms. A newly developed kinase assay revealed that DMPK is a Lys/Arg-directed kinase. Individual DMPK isoforms displayed comparable transphosphorylation activity and sequence preference for peptide substrates. However, DMPK autophosphorylation and phosphorylation of MYPT1 (as putative in vivo target of DMPK), were dependent on presence of an alternatively spliced VSGGG motif and the nature of the C terminus. In-gel effects of the VSGGG motif on the migration behavior of full-length kinase provide evidence for a model in which this motif mediates 3-D-conformational changes in DMPK isoforms. Finally, different C termini endow DMPK with the ability to bind to either endoplasmic reticulum or mitochondria or to adopt a cytosolic location. Our results suggest that DMPK isoforms have cell-type and location dependent substrate specificities with a role in organellar and cytoarchitectural dynamics. PMID- 12897126 TI - Degradation of normal mRNA in the nucleus of Saccharomyces cerevisiae. AB - A nuclear mRNA degradation (DRN) system was identified from analysis of mRNA turnover rates in nup116-Delta strains of Saccharomyces cerevisiae lacking the ability to export all RNAs, including poly(A) mRNAs, at the restrictive temperature. Northern blotting, in situ hybridization, and blocking transcription with thiolutin in nup116-delta strains revealed a rapid degradation of mRNAs in the nucleus that was suppressed by the rrp6-delta, rai1-delta, and cbc1-delta deletions, but not by the upf1-delta deletion, suggesting that DRN requires Rrp6p, a 3'-to-5' nuclear exonuclease, the Rat1p, a 5'-to-3' nuclear exonuclease, and Cbc1p, a component of CBC, the nuclear cap binding complex, which may direct the mRNAs to the site of degradation. We propose that certain normal mRNAs retained in the nucleus are degraded by the DRN system, similar to degradation of transcripts with 3' end formation defects in certain mutants. PMID- 12897127 TI - Identification of a unique core domain of par-4 sufficient for selective apoptosis induction in cancer cells. AB - Recent studies indicated that the leucine zipper domain protein Par-4 induces apoptosis in certain cancer cells by activation of the Fas prodeath pathway and coparallel inhibition of NF-kappaB transcriptional activity. However, the intracellular localization or functional domains of Par-4 involved in apoptosis remained unknown. In the present study, structure-function analysis indicated that inhibition of NF-kappaB activity and apoptosis is dependent on Par-4 translocation to the nucleus via a bipartite nuclear localization sequence, NLS2. Cancer cells that were resistant to Par-4-induced apoptosis retained Par-4 in the cytoplasm. Interestingly, a 59-amino-acid core that included NLS2 but not the C terminal leucine zipper domain was necessary and sufficient to induce Fas pathway activation, inhibition of NF-kappaB activity, and apoptosis. Most important, this core domain had an expanded target range for induction of apoptosis, extending to previously resistant cancer cells but not to normal cells. These findings have identified a unique death-inducing domain selective for apoptosis induction in cancer cells (SAC domain) which holds promise for identifying key differences between cancer and normal cells and for molecular therapy of cancer. PMID- 12897128 TI - p21-Activated kinase 5 (Pak5) localizes to mitochondria and inhibits apoptosis by phosphorylating BAD. AB - Pak5 is the most recently identified and least understood member of the p21 activated kinase (Pak) family. This kinase is known to promote neurite outgrowth in vitro, but its localization, substrates, and effects on cell survival have not been reported. We show here that Pak5 has unique properties that distinguish it from all other members of the Pak family. First, Pak5, unlike Pak1, cannot complement an STE20 mutation in Saccharomyces cerevisiae. Second, Pak5 binds to the GTPases Cdc42 and Rac, but these GTPases do not regulate Pak5 kinase activity, which is constitutive and stronger than any other Pak. Third, Pak5 prevents apoptosis induced by camptothecin and C2-ceramide by phosphorylating BAD on Ser-112 in a protein kinase A-independent manner and prevents the localization of BAD to mitochondria, thereby inhibiting the apoptotic cascade that leads to apoptosis. Finally, we show that Pak5 itself is constitutively localized to mitochondria, and that this localization is independent of kinase activity or Cdc42 binding. These features make Pak5 unique among the Pak family and suggest that it plays an important role in apoptosis through BAD phosphorylation. PMID- 12897129 TI - deltaNp73 facilitates cell immortalization and cooperates with oncogenic Ras in cellular transformation in vivo. AB - TP73, despite significant homology to TP53, is not a classic tumor suppressor gene, since it exhibits upregulation of nonmutated products in human tumors and lacks a tumor phenotype in p73-deficient mice. We recently reported that an N terminally truncated isoform, DeltaNp73, is upregulated in breast and gynecological cancers. We further showed that DeltaNp73 is a potent transdominant inhibitor of wild-type p53 and TAp73 in cultured human tumor cells by efficiently counteracting their target gene transactivations, apoptosis, and growth suppression functions (A. I. Zaika et al., J. Exp. Med. 6:765-780, 2002). Although these data strongly suggest oncogenic properties of DeltaNp73, this can only be directly shown in primary cells. We report here that DeltaNp73 confers resistance to spontaneous replicative senescence of primary mouse embryo fibroblasts (MEFs) and immortalizes MEFs at a 1,000-fold-higher frequency than occurs spontaneously. DeltaNp73 cooperates with cMyc and E1A in promoting primary cell proliferation and colony formation and compromises p53-dependent MEF apoptosis. Importantly, DeltaNp73 rescues Ras-induced senescence. Moreover, DeltaNp73 cooperates with oncogenic Ras in transforming primary fibroblasts in vitro and in inducing MEF-derived fibrosarcomas in vivo in nude mice. Wild-type p53 is likely a major target of DeltaNp73 inhibition in primary fibroblasts since deletion of p53 or its requisite upstream activator ARF abrogates the growth promoting effect of DeltaNp73. Taken together, DeltaNp73 behaves as an oncogene that targets p53 that might explain why DeltaNp73 upregulation may be selected for during tumorigenesis of human cancers. PMID- 12897130 TI - Silencing of the novel p53 target gene Snk/Plk2 leads to mitotic catastrophe in paclitaxel (taxol)-exposed cells. AB - Loss of p53 sensitizes to antimicrotubule agents in human tumor cells, but little is known about its role during mitosis. We have identified the Polo-like kinase family member serum inducible kinase (Snk/Plk2) as a novel p53 target gene. Snk/Plk2 mutagenesis demonstrated that its kinase activity is negatively regulated by its C terminus. Small interfering RNA (siRNA)-mediated Snk/Plk2 silencing in the presence of the mitotic poisons paclitaxel (Taxol) or nocodazole significantly increased apoptosis, similar to p53 mutations, which confer paclitaxel sensitivity. Furthermore, we have demonstrated that the apoptosis due to silencing of Snk/Plk2 in the face of spindle damage occurs in mitotic cells and not in cells that have progressed to a G(1)-like state without dividing. Since siRNA directed against Snk/Plk2 promoted death of paclitaxel-treated cells in mitosis, we envision a mitotic checkpoint wherein p53-dependent activation of Snk/Plk2 prevents mitotic catastrophe following spindle damage. Finally, these studies suggest that disruption of Snk/Plk2 may be of therapeutic value in sensitizing paclitaxel-resistant tumors. PMID- 12897131 TI - Role of mammalian Rad54 in telomere length maintenance. AB - The homologous recombination (HR) DNA repair pathway participates in telomere length maintenance in yeast but its putative role at mammalian telomeres is unknown. Mammalian Rad54 is part of the HR machinery, and Rad54-deficient mice show a reduced HR capability. Here, we show that Rad54-deficient mice also show significantly shorter telomeres than wild-type controls, indicating that Rad54 activity plays an essential role in telomere length maintenance in mammals. Rad54 deficiency also resulted in an increased frequency of end-to-end chromosome fusions involving telomeres compared to the controls, suggesting a putative role of Rad54 in telomere capping. Finally, the study of mice doubly deficient for Rad54 and DNA-PKcs showed that telomere fusions due to DNA-PKcs deficiency were not rescued in the absence of Rad54, suggesting that they are not mediated by Rad54 activity. PMID- 12897132 TI - Regulation of Notch signaling by a novel mechanism involving suppressor of hairless stability and carboxyl terminus-truncated notch. AB - Different amounts of Suppressor of Hairless (SuH)-dependent Notch (N) signaling is often used during animal development to produce two different tissues from a population of equipotent cells. During Drosophila melanogaster embryogenesis, cells with high amounts of this signaling differentiate the larval epidermis whereas cells with low amounts, or none, differentiate the central nervous system (CNS). The mechanism by which SuH-dependent N signaling is increased or decreased in these different cells is obscure. The developing epidermis is known to get enriched for the full-length N (NFull) and the developing CNS for the carboxyl terminus-truncated N (NdeltaCterm). Results described here indicate that this differential accumulation of N receptors is part of a mechanism that would promote SuH-dependent N signaling in the developing epidermis but suppress it in the developing CNS. This mechanism involves SuH-dependent stability of NFull, NFull-dependent accumulation of SuH, stage specific stability of SuH, and NdeltaCterm-dependent loss of SuH and NFull. PMID- 12897133 TI - Establishment and maintenance of genomic methylation patterns in mouse embryonic stem cells by Dnmt3a and Dnmt3b. AB - We have previously shown that the DNA methyltransferases Dnmt3a and Dnmt3b carry out de novo methylation of the mouse genome during early postimplantation development and of maternally imprinted genes in the oocyte. In the present study, we demonstrate that Dnmt3a and Dnmt3b are also essential for the stable inheritance, or "maintenance," of DNA methylation patterns. Inactivation of both Dnmt3a and Dnmt3b in embryonic stem (ES) cells results in progressive loss of methylation in various repeats and single-copy genes. Interestingly, introduction of the Dnmt3a, Dnmt3a2, and Dnmt3b1 isoforms back into highly demethylated mutant ES cells restores genomic methylation patterns; these isoforms appear to have both common and distinct DNA targets, but they all fail to restore the maternal methylation imprints. In contrast, overexpression of Dnmt1 and Dnmt3b3 failed to restore DNA methylation patterns due to their inability to catalyze de novo methylation in vivo. We also show that hypermethylation of genomic DNA by Dnmt3a and Dnmt3b is necessary for ES cells to form teratomas in nude mice. These results indicate that genomic methylation patterns are determined partly through differential expression of different Dnmt3a and Dnmt3b isoforms. PMID- 12897134 TI - Stem-loop IV of tetrahymena telomerase RNA stimulates processivity in trans. AB - Telomerase is a ribonucleoprotein enzyme responsible for the addition of telomeres onto the ends of chromosomes. Short or dysfunctional telomeres can lead to cell growth arrest, apoptosis, and genomic instability. Telomerase uses its RNA subunit to copy a short template region for telomere synthesis. To probe for regions of Tetrahymena telomerase RNA essential for function, we assayed 27 circularly permuted RNA deletions for telomerase in vitro activity and binding to the telomerase reverse transcriptase catalytic protein subunit. We found that stem-loop IV is required for wild-type telomerase activity in vitro and will stimulate processivity when added in trans. PMID- 12897135 TI - Potential role for ADAM15 in pathological neovascularization in mice. AB - ADAM15 (named for a disintegrin and metalloprotease 15, metargidin) is a membrane anchored glycoprotein that has been implicated in cell-cell or cell-matrix interactions and in the proteolysis of molecules on the cell surface or extracellular matrix. To characterize the potential roles of ADAM15 during development and in adult mice, we analyzed its expression pattern by mRNA in situ hybridization and generated mice carrying a targeted deletion of ADAM15 (adam15( /-) mice). A high level of expression of ADAM15 was found in vascular cells, the endocardium, hypertrophic cells in developing bone, and specific areas of the hippocampus and cerebellum. However, despite the pronounced expression of ADAM15 in these tissues, no major developmental defects or pathological phenotypes were evident in adam15(-/-) mice. The elevated levels of ADAM15 in endothelial cells prompted an evaluation of its role in neovascularization. In a mouse model for retinopathy of prematurity, adam15(-/-) mice had a major reduction in neovascularization compared to wild-type controls. Furthermore, the size of tumors resulting from implanted B16F0 mouse melanoma cells was significantly smaller in adam15(-/-) mice than in wild-type controls. Since ADAM15 does not appear to be required for developmental angiogenesis or for adult homeostasis, it may represent a novel target for the design of inhibitors of pathological neovascularization. PMID- 12897136 TI - The Drosophila dysfusion basic helix-loop-helix (bHLH)-PAS gene controls tracheal fusion and levels of the trachealess bHLH-PAS protein. AB - The development of the mature insect trachea requires a complex series of cellular events, including tracheal cell specification, cell migration, tubule branching, and tubule fusion. Here we describe the identification of the Drosophila melanogaster dysfusion gene, which encodes a novel basic helix-loop helix (bHLH)-PAS protein conserved between Caenorhabditis elegans, insects, and humans, and controls tracheal fusion events. The Dysfusion protein functions as a heterodimer with the Tango bHLH-PAS protein in vivo to form a putative DNA binding complex. The dysfusion gene is expressed in a variety of embryonic cell types, including tracheal-fusion, leading-edge, foregut atrium cells, nervous system, hindgut, and anal pad cells. RNAi experiments indicate that dysfusion is required for dorsal branch, lateral trunk, and ganglionic branch fusion but not for fusion of the dorsal trunk. The escargot gene, which is also expressed in fusion cells and is required for tracheal fusion, precedes dysfusion expression. Analysis of escargot mutants indicates a complex pattern of dysfusion regulation, such that dysfusion expression is dependent on escargot in the dorsal and ganglionic branches but not the dorsal trunk. Early in tracheal development, the Trachealess bHLH-PAS protein is present at uniformly high levels in all tracheal cells, but since the levels of Dysfusion rise in wild-type fusion cells, the levels of Trachealess in fusion cells decline. The downregulation of Trachealess is dependent on dysfusion function. These results suggest the possibility that competitive interactions between basic helix-loop-helix-PAS proteins (Dysfusion, Trachealess, and possibly Similar) may be important for the proper development of the trachea. PMID- 12897137 TI - A prokaryotic condensin/cohesin-like complex can actively compact chromosomes from a single position on the nucleoid and binds to DNA as a ring-like structure. AB - We show that Bacillus subtilis SMC (structural maintenance of chromosome protein) localizes to discrete foci in a cell cycle-dependent manner. Early in the cell cycle, SMC moves from the middle of the cell toward opposite cell poles in a rapid and dynamic manner and appears to interact with different regions on the chromosomes during the cell cycle. SMC colocalizes with its interacting partners, ScpA and ScpB, and the specific localization of SMC depends on both Scp proteins, showing that all three components of the SMC complex are required for proper localization. Cytological and biochemical experiments showed that dimeric ScpB stabilized the binding of ScpA to the SMC head domains. Purified SMC showed nonspecific binding to double-stranded DNA, independent of Scp proteins or ATP, and was retained on DNA after binding to closed DNA but not to linear DNA. The SMC head domains and hinge region did not show strong DNA binding activity, suggesting that the coiled-coil regions in SMC mediate an association with DNA and that SMC binds to DNA as a ring-like structure. The overproduction of SMC resulted in global chromosome compaction, while SMC was largely retained in bipolar foci, suggesting that the SMC complex forms condensation centers that actively affect global chromosome compaction from a defined position on the nucleoid. PMID- 12897138 TI - Phosphorylation of the alpha subunit of eukaryotic initiation factor 2 is required for activation of NF-kappaB in response to diverse cellular stresses. AB - Nuclear factor kappaB (NF-kappaB) serves to coordinate the transcription of genes in response to diverse environmental stresses. In this report we show that phosphorylation of the alpha subunit of eukaryotic initiation factor 2 (eIF2) is fundamental to the process by which many stress signals activate NF-kappaB. Phosphorylation of this translation factor is carried out by a family of protein kinases that each respond to distinct stress conditions. During impaired protein folding and assembly in the endoplasmic reticulum (ER), phosphorylation of eIF2alpha by PEK (Perk or EIF2AK3) is essential for induction of NF-kappaB transcriptional activity. The mechanism by which NF-kappaB is activated during ER stress entails the release, but not the degradation, of the inhibitory protein IkappaB. During amino acid deprivation, phosphorylation of eIF2alpha by GCN2 (EIF2AK4) signals the activation of NF-kappaB. Furthermore, inhibition of general translation or transcription by cycloheximide and actinomycin D, respectively, elicits the eIF2alpha phosphorylation required for induction of NF-kappaB. Together, these studies suggest that eIF2alpha kinases monitor and are activated by a range of stress conditions that affect transcription and protein synthesis and assembly, and the resulting eIFalpha phosphorylation is central to activation of the NF-kappaB. The absence of NF-kappaB-mediated transcription and its antiapoptotic function provides an explanation for why eIF2alpha kinase deficiency in diseases such as Wolcott-Rallison syndrome leads to cellular apoptosis and disease. PMID- 12897140 TI - HoxB5 is an upstream transcriptional switch for differentiation of the vascular endothelium from precursor cells. AB - Endothelial cells differentiate from mesoderm-derived precursors to initiate the earliest events in vascular development. Although the signaling events that regulate the successive steps of vascular development are known in some detail, the transcriptional processes that regulate the first steps in vasculogenesis are not well defined. We have studied the regulatory mechanisms of flk1 expression as a model to understand the upstream events in endothelial cell differentiation, since flk1 is the earliest marker of endothelial precursors. Using a variety of biochemical approaches, we identified a cis-acting element in the first intron of the flk1 gene that is required for endothelium-dependent expression in transgenic reporter gene assays. Using the yeast one-hybrid system, we identified HoxB5 as the transcription factor that binds this cis-acting element, the HoxB5-binding element (HBE). HoxB5 mRNA colocalized with flk1 expression in differentiating embryoid bodies, and HoxB5 potently transactivated the flk1 promoter in an HBE dependent fashion in transient-transfection assays. Overexpression of HoxB5 led to expansion of flk1(+) angioblasts in differentiating embryoid bodies and increased the number of PECAM (platelet-endothelial cell adhesion molecule) positive primitive blood vessels. HoxB5 is necessary and sufficient to activate the cell-intrinsic events that regulate the differentiation of angioblasts and mature endothelial cells from their mesoderm-derived precursors. PMID- 12897139 TI - BMPER, a novel endothelial cell precursor-derived protein, antagonizes bone morphogenetic protein signaling and endothelial cell differentiation. AB - The development of endothelial cell precursors is essential for vasculogenesis. We screened for differentially expressed transcripts in endothelial cell precursors in developing mouse embryoid bodies. We cloned a complete cDNA encoding a protein that contains an amino-terminal signal peptide, five cysteine rich domains, a von Willebrand D domain, and a trypsin inhibitor domain. We termed this protein BMPER (bone morphogenetic protein [BMP]-binding endothelial cell precursor-derived regulator). BMPER is specifically expressed in flk-1 positive cells and parallels the time course of flk-1 induction in these cells. In situ hybridization in mouse embryos demonstrates dorsal midline staining and staining of the aorto-gonadal-mesonephric region, which is known to host vascular precursor cells. BMPER is a secreted protein that directly interacts with BMP2, BMP4, and BMP6 and antagonizes BMP4-dependent Smad5 activation. In Xenopus embryos, ventral injection of BMPER mRNA results in axis duplication and downregulation of the expression of Xvent-1 (downstream target of Smad signaling). In an embryoid body differentiation assay, BMP4-dependent differentiation of endothelial cells in embryoid bodies is also antagonized by BMPER. Taken together, our data indicate that BMPER is a novel BMP-binding protein that is expressed by endothelial cell precursors, has BMP-antagonizing activity, and may play a role in endothelial cell differentiation by modulating local BMP activity. PMID- 12897141 TI - The N and C termini of the splice variants of the human mitogen-activated protein kinase-interacting kinase Mnk2 determine activity and localization. AB - The cap-binding eukaryotic initiation factor eIF4E is phosphorylated by the mitogen-activated protein (MAP) kinase-interacting kinases (Mnk's). Three forms of the Mnk's exist in human cells: Mnk1, Mnk2a, and Mnk2b. These last two are derived from the same gene by alternative splicing and differ only at their C termini. While Mnk2a contains a MAP kinase-binding site in this region, Mnk2b lacks such a sequence and is much less readily activated by MAP kinases in vitro. Expression of Mnk2b in mammalian cells leads to increased phosphorylation of eIF4E, showing that it acts as an eIF4E kinase in vivo. While Mnk2a is cytoplasmic, a substantial amount of Mnk2b is found in the nucleus. Both enzymes contain a stretch of basic residues in their N termini that plays a role in binding to eIF4G and functions as a nuclear localization signal. Binding of eIF4G or nuclear import appears to be regulated by the C terminus of Mnk2a. Furthermore, the MAP kinase-binding site of Mnk2a regulates nuclear entry. Within the nucleus, Mnk2b and certain variants of Mnk2a that are present in the nucleus colocalize with the promyelocytic leukemia protein PML, which also binds to eIF4E. PMID- 12897142 TI - Pathways of DNA double-strand break repair during the mammalian cell cycle. AB - Little is known about the quantitative contributions of nonhomologous end joining (NHEJ) and homologous recombination (HR) to DNA double-strand break (DSB) repair in different cell cycle phases after physiologically relevant doses of ionizing radiation. Using immunofluorescence detection of gamma-H2AX nuclear foci as a novel approach for monitoring the repair of DSBs, we show here that NHEJ defective hamster cells (CHO mutant V3 cells) have strongly reduced repair in all cell cycle phases after 1 Gy of irradiation. In contrast, HR-defective CHO irs1SF cells have a minor repair defect in G(1), greater impairment in S, and a substantial defect in late S/G(2). Furthermore, the radiosensitivity of irs1SF cells is slight in G(1) but dramatically higher in late S/G(2), while V3 cells show high sensitivity throughout the cell cycle. These findings show that NHEJ is important in all cell cycle phases, while HR is particularly important in late S/G(2), where both pathways contribute to repair and radioresistance. In contrast to DSBs produced by ionizing radiation, DSBs produced by the replication inhibitor aphidicolin are repaired entirely by HR. irs1SF, but not V3, cells show hypersensitivity to aphidicolin treatment. These data provide the first evaluation of the cell cycle-specific contributions of NHEJ and HR to the repair of radiation-induced versus replication-associated DSBs. PMID- 12897143 TI - Caspase 3-mediated inactivation of rac GTPases promotes drug-induced apoptosis in human lymphoma cells. AB - The Rac members of the Rho family GTPases control signaling pathways that regulate diverse cellular activities, including cytoskeletal organization, gene transcription, and cell transformation. Rac is implicated in apoptosis, but little is known about the mechanism by which it responds to apoptotic stimuli. Here we demonstrate that endogenous Rac GTPases are caspase 3 substrates that are cleaved in human lymphoma cells during drug-induced apoptosis. Cleavage of Rac1 occurs at two unconventional caspase 3 sites, VVGD11/G and VMVD47/G, and results in inactivation of the GTPase and effector functions of the protein (binding to the p21-activated protein kinase PAK1). Expression of caspase 3-resistant Rac1 mutants in the cells suppresses drug-induced apoptosis. Thus, proteolytic inactivation of Rac GTPases represents a novel, irreversible mechanism of Rac downregulation that allows maximal cell death following drug treatment. PMID- 12897144 TI - Activation of the phosphatidylinositol 3-kinase/protein kinase Akt pathway mediates nitric oxide-induced endothelial cell migration and angiogenesis. AB - To test the hypothesis that the phosphatidylinositol 3-kinase (PI3 kinase)/protein kinase Akt signaling pathway is involved in nitric oxide (NO) induced endothelial cell migration and angiogenesis, we treated human and bovine endothelial cells with NO donors, S-nitroso-L-glutathione (GSNO) and S-nitroso-N penicillamine (SNAP). Both GSNO and SNAP increased Akt phosphorylation and activity, which were blocked by cotreatment with the PI3 kinase inhibitor wortmannin. The mechanism was due to the activation of soluble guanylyl cyclase because 8-bromo-cyclic GMP activated PI3 kinase and the soluble guanylyl cyclase inhibitor 1H-[1,2,4]oxadiazolo[4,3-alpha]quinoxalin-1-one (ODQ) blocked NO induced PI3 kinase activity. Indeed, transfection with adenovirus containing endothelial cell NO synthase (eNOS) or protein kinase G (PKG) increased endothelial cell migration, which was inhibited by cotransfection with a dominant negative mutant of PI3 kinase (dnPI3 kinase). In a rat model of hind limb ischemia, adenovirus-mediated delivery of human eNOS cDNA in adductor muscles resulted in time-dependent expression of recombinant eNOS, which was accompanied by significant increases in regional blood perfusion and capillary density. Coinjection of adenovirus carrying dnPI3 kinase abolished neovascularization in ischemic hind limb induced by eNOS gene transfer. These findings indicate that NO promotes endothelial cell migration and neovascularization via cGMP-dependent activation of PI3 kinase and suggest that this pathway is important in mediating NO-induced angiogenesis. PMID- 12897145 TI - Mouse mammary tumor virus c-rel transgenic mice develop mammary tumors. AB - Amplification, overexpression, or rearrangement of the c-rel gene, encoding the c Rel NF-kappaB subunit, has been reported in solid and hematopoietic malignancies. For example, many primary human breast cancer tissue samples express high levels of nuclear c-Rel. While the Rev-T oncogene v-rel causes tumors in birds, the ability of c-Rel to transform in vivo has not been demonstrated. To directly test the role of c-Rel in breast tumorigenesis, mice were generated in which overexpression of mouse c-rel cDNA was driven by the hormone-responsive mouse mammary tumor virus long terminal repeat (MMTV-LTR) promoter, and four founder lines identified. In the first cycle of pregnancy, the expression of transgenic c rel mRNA was observed, and levels of c-Rel protein were increased in the mammary gland. Importantly, 31.6% of mice developed one or more mammary tumors at an average age of 19.9 months. Mammary tumors were of diverse histology and expressed increased levels of nuclear NF-kappaB. Analysis of the composition of NF-kappaB complexes in the tumors revealed aberrant nuclear expression of multiple subunits, including c-Rel, p50, p52, RelA, RelB, and the Bcl-3 protein, as observed previously in human primary breast cancers. Expression of the cancer related NF-kappaB target genes cyclin D1, c-myc, and bcl-xl was significantly increased in grossly normal transgenic mammary glands starting the first cycle of pregnancy and increased further in mammary carcinomas compared to mammary glands from wild-type mice or virgin transgenic mice. In transient transfection analysis in untransformed breast epithelial cells, c-Rel-p52 or -p50 heterodimers either potently or modestly induced cyclin D1 promoter activity, respectively. Lastly, stable overexpression of c-Rel resulted in increased cyclin D1 and NF-kappaB p52 and p50 subunit protein levels. These results indicate for the first time that dysregulated expression of c-Rel, as observed in breast cancers, is capable of contributing to mammary tumorigenesis. PMID- 12897146 TI - Xeroderma pigmentosum group A protein loads as a separate factor onto DNA lesions. AB - Nucleotide excision repair (NER) is the main DNA repair pathway in mammals for removal of UV-induced lesions. NER involves the concerted action of more than 25 polypeptides in a coordinated fashion. The xeroderma pigmentosum group A protein (XPA) has been suggested to function as a central organizer and damage verifier in NER. How XPA reaches DNA lesions and how the protein is distributed in time and space in living cells are unknown. Here we studied XPA in vivo by using a cell line stably expressing physiological levels of functional XPA fused to green fluorescent protein and by applying quantitative fluorescence microscopy. The majority of XPA moves rapidly through the nucleoplasm with a diffusion rate different from those of other NER factors tested, arguing against a preassembled XPA-containing NER complex. DNA damage induced a transient ( approximately 5-min) immobilization of maximally 30% of XPA. Immobilization depends on XPC, indicating that XPA is not the initial lesion recognition protein in vivo. Moreover, loading of replication protein A on NER lesions was not dependent on XPA. Thus, XPA participates in NER by incorporation of free diffusing molecules in XPC-dependent NER-DNA complexes. This study supports a model for a rapid consecutive assembly of free NER factors, and a relatively slow simultaneous disassembly, after repair. PMID- 12897147 TI - Cotranscriptional recruitment of the U1 snRNP to intron-containing genes in yeast. AB - Evidence that pre-mRNA processing events are temporally and, in some cases, mechanistically coupled to transcription has led to the proposal that RNA polymerase II (Pol II) recruits pre-mRNA splicing factors to active genes. Here we address two key questions raised by this proposal: (i) whether the U1 snRNP, which binds to the 5' splice site of each intron, is recruited cotranscriptionally in vivo and, (ii) if so, where along the length of active genes the U1 snRNP is concentrated. Using chromatin immunoprecipitation (ChIP) in yeast, we show that elevated levels of the U1 snRNP were specifically detected in gene regions containing introns and downstream of introns but not along the length of intronless genes. In contrast to capping enzymes, which bind directly to Pol II, the U1 snRNP was poorly detected in promoter regions, except in genes harboring promoter-proximal introns. Detection of the U1 snRNP was dependent on RNA synthesis and was abolished by intron removal. Microarray analysis revealed that intron-containing genes were preferentially selected by ChIP with the U1 snRNP. Thus, U1 snRNP accumulation at genes correlated with the presence and position of introns, indicating that introns are necessary for cotranscriptional U1 snRNP recruitment and/or retention. PMID- 12897148 TI - Promoter-specific roles for liver X receptor/corepressor complexes in the regulation of ABCA1 and SREBP1 gene expression. AB - Liver X receptors (LXRs) regulate the expression of genes involved in cholesterol and fatty acid homeostasis, including the genes for ATP-binding cassette transporter A1 (ABCA1) and sterol response element binding protein 1 (SREBP1). Loss of LXR leads to derepression of the ABCA1 gene in macrophages and the intestine, while the SREBP1c gene remains transcriptionally silent. Here we report that high-density-lipoprotein (HDL) cholesterol levels are increased in LXR-deficient mice, suggesting that derepression of ABCA1 and possibly other LXR target genes in selected tissues is sufficient to result in enhanced HDL biogenesis at the whole-body level. We provide several independent lines of evidence indicating that the repressive actions of LXRs are dependent on interactions with the nuclear receptor corepressor (NCoR) and the silencing mediator of retinoic acid and thyroid hormone receptors (SMRT). While dissociation of NCoR and SMRT results in derepression of the ABCA1 gene in macrophages, it is not sufficient for derepression of the SREBP1c gene. These findings reveal differential requirements for corepressors in the regulation of genes involved in cholesterol and fatty acid homeostasis and raise the possibility that these interactions may be exploited to develop synthetic ligands that selectively modulate LXR actions in vivo. PMID- 12897149 TI - HSP27 is a ubiquitin-binding protein involved in I-kappaBalpha proteasomal degradation. AB - HSP27 is an ATP-independent chaperone that confers protection against apoptosis through various mechanisms, including a direct interaction with cytochrome c. Here we show that HSP27 overexpression in various cell types enhances the degradation of ubiquitinated proteins by the 26S proteasome in response to stressful stimuli, such as etoposide or tumor necrosis factor alpha (TNF-alpha). We demonstrate that HSP27 binds to polyubiquitin chains and to the 26S proteasome in vitro and in vivo. The ubiquitin-proteasome pathway is involved in the activation of transcription factor NF-kappaB by degrading its main inhibitor, I kappaBalpha. HSP27 overexpression increases NF-kappaB nuclear relocalization, DNA binding, and transcriptional activity induced by etoposide, TNF-alpha, and interleukin 1beta. HSP27 does not affect I-kappaBalpha phosphorylation but enhances the degradation of phosphorylated I-kappaBalpha by the proteasome. The interaction of HSP27 with the 26S proteasome is required to activate the proteasome and the degradation of phosphorylated I-kappaBalpha. A protein complex that includes HSP27, phosphorylated I-kappaBalpha, and the 26S proteasome is formed. Based on these observations, we propose that HSP27, under stress conditions, favors the degradation of ubiquitinated proteins, such as phosphorylated I-kappaBalpha. This novel function of HSP27 would account for its antiapoptotic properties through the enhancement of NF-kappaB activity. PMID- 12897150 TI - Stimulation of cell proliferation by endosomal epidermal growth factor receptor as revealed through two distinct phases of signaling. AB - Strong evidence indicates that endosome-localized epidermal growth factor receptor (EGFR) plays an important role in cell signaling. However, elimination of endosomal signaling does not attenuate EGF-induced physiological outcomes, arguing against physiological relevance. Recently we established a system to specifically activate endosome-associated EGFR in the absence of any plasma membrane activation of EGFR and showed that endosomal EGFR signaling is sufficient to support cell survival. However, this pure endosomal signaling of EGFR does not stimulate cell proliferation, because EGFR only remained activated for less than 2 h following its stimulation at endosomes, while DNA synthesis generally requires growth factor exposure for 8 h or more. Here we report that the prolonged requirement for EGF to stimulate epithelial cell proliferation can be substituted for with two short pulses of EGF. By combining the two short pulses of EGF stimulation with our previously established method to generate endosomal EGFR signaling, we are able to generate two pulses of endosomal EGFR signaling. In this way, we demonstrated that two pulses of endosomal EGFR signaling are sufficient to stimulate cell proliferation. The first pulse of EGFR signaling induces exit from quiescence into G(1) phase and appears to render cells responsive to subsequent mitogenic stimulus. This second pulse, required several hours later, drives cells through the restriction point of late G(1) and into S phase. We further showed that the two pulses of endosomal EGFR signaling engaged cell cycle machinery the same way as the two pulses of standard EGFR signaling. Moreover, two pulses of endosomal EGFR signaling stimulated downstream signaling cascades in a similar way to the two pulses of standard EGFR activation. The data therefore demonstrate that signals transduced from internalized EGFR, with or without a contribution from the plasma membrane, fully satisfy the physiological requirements for S-phase entry. PMID- 12897151 TI - Human mitochondrial transcription factor B1 interacts with the C-terminal activation region of h-mtTFA and stimulates transcription independently of its RNA methyltransferase activity. AB - A significant advancement in understanding mitochondrial gene expression is the recent identification of two new human mitochondrial transcription factors, h mtTFB1 and h-mtTFB2. Both proteins stimulate transcription in collaboration with the high-mobility group box transcription factor, h-mtTFA, and are homologous to rRNA methyltransferases. In fact, the dual-function nature of h-mtTFB1 was recently demonstrated by its ability to methylate a conserved rRNA substrate. Here, we demonstrate that h-mtTFB1 binds h-mtTFA both in HeLa cell mitochondrial extracts and in direct-binding assays via an interaction that requires the C terminal tail of h-mtTFA, a region necessary for transcriptional activation. In addition, point mutations in conserved methyltransferase motifs of h-mtTFB1 revealed that it stimulates transcription in vitro independently of S adenosylmethionine binding and rRNA methyltransferase activity. Furthermore, one mutation (G65A) eliminated the ability of h-mtTFB1 to bind DNA yet did not affect transcriptional activation. These results, coupled with the observation that h mtTFB1 and human mitochondrial RNA (h-mtRNA) polymerase can also be coimmunoprecipitated, lead us to propose a model in which h-mtTFA demarcates mitochondrial promoter locations and where h-mtTFB proteins bridge an interaction between the C-terminal tail of h-mtTFA and mtRNA polymerase to facilitate specific initiation of transcription. Altogether, these data provide important new insight into the mechanism of transcription initiation in human mitochondria and indicate that the dual functions of h-mtTFB1 can be separated. PMID- 12897152 TI - A novel mechanism for Wnt activation of canonical signaling through the LRP6 receptor. AB - LDL receptor-related protein 6 (LRP6) is a Wnt coreceptor in the canonical signaling pathway, which plays essential roles in embryonic development. We demonstrate here that wild-type LRP6 forms an inactive dimer through interactions mediated by epidermal growth factor repeat regions within the extracellular domain. A truncated LRP6 comprising its transmembrane and cytoplasmic domains is expressed as a constitutively active monomer whose signaling ability is inhibited by forced dimerization. Conversely, Wnts are shown to activate canonical signaling through LRP6 by inducing an intracellular conformational switch which relieves allosteric inhibition imposed on the intracellular domains. Thus, Wnt canonical signaling through LRP6 establishes a novel mechanism for receptor activation which is opposite to the general paradigm of ligand-induced receptor oligomerization. PMID- 12897153 TI - Autophosphorylation of the catalytic subunit of the DNA-dependent protein kinase is required for efficient end processing during DNA double-strand break repair. AB - The DNA-dependent protein kinase (DNA-PK) plays an essential role in nonhomologous DNA end joining (NHEJ) by initially recognizing and binding to DNA breaks. We have shown that in vitro, purified DNA-PK undergoes autophosphorylation, resulting in loss of activity and disassembly of the kinase complex. Thus, we have suggested that autophosphorylation of the DNA-PK catalytic subunit (DNA-PKcs) may be critical for subsequent steps in DNA repair. Recently, we defined seven autophosphorylation sites within DNA-PKcs. Six of these are tightly clustered within 38 residues of the 4,127-residue protein. Here, we show that while phosphorylation at any single site within the major cluster is not critical for DNA-PK's function in vivo, mutation of several sites abolishes the ability of DNA-PK to function in NHEJ. This is not due to general defects in DNA PK activity, as studies of the mutant protein indicate that its kinase activity and ability to form a complex with DNA-bound Ku remain largely unchanged. However, analysis of rare coding joints and ends demonstrates that nucleolytic end processing is dramatically reduced in joints mediated by the mutant DNA-PKcs. We therefore suggest that autophosphorylation within the major cluster mediates a conformational change in the DNA-PK complex that is critical for DNA end processing. However, autophosphorylation at these sites may not be sufficient for kinase disassembly. PMID- 12897154 TI - TRAF2 exerts its antiapoptotic effect by regulating the expression of Kruppel like factor LKLF. AB - Tumor necrosis factor receptor (TNFR)-associated factor 2 (TRAF2) is one of the key factors that mediate TNF signaling. The deletion of TRAF2 renders cells more sensitive to TNF-induced apoptosis. Although TRAF2 is known to be required for TNF-induced JNK and NF-kappaB activation, the underlying mechanism of the increased sensitivity of TRAF2 null cells (TRAF2(-/-)) to TNF-induced apoptosis is not fully understood. To study the underlying mechanism, we examined the difference in gene expression between TRAF2(-/-) and wild-type fibroblast cells by using microarray technology. We found that one of the genes whose expression was dramatically decreased in TRAF2(-/-) cells was the lung Kruppel-like factor (LKLF). Our results indicate that the expression of LKLF requires TRAF2 but is independent of TNF signaling. Although it appears that TRAF2 regulates the expression of the LKLF gene at the transcription level, TRAF2 does not function as a transcription factor itself. Our results suggest that TRAF2 regulates LKLF expression through the mitogen-activated protein kinase p38 pathway. More importantly, ectopic expression of LKLF in TRAF2(-/-) cells protected cells against TNF-induced apoptosis. These results reveal a novel aspect of TRAF2 function: by regulating the expression of genes, such as LKLF, TRAF2 controls cell sensitivity to apoptosis. PMID- 12897155 TI - Structural environment dictates the biological significance of heme-responsive motifs and the role of Hsp90 in the activation of the heme activator protein Hap1. AB - Heme-responsive motifs (HRMs) mediate heme regulation of diverse regulatory proteins. The heme activator protein Hap1 contains seven HRMs, but only one of them, HRM7, is essential for heme activation of Hap1. To better understand the molecular basis underlying the biological significance of HRMs, we examined the effects of various mutations of HRM7 on Hap1. We found that diverse mutations of HRM7 significantly diminished the extent of Hap1 activation by heme and moderately enhanced the interaction of Hap1 with Hsp90. Furthermore, deletions of nonregulatory sequences completely abolished heme activation of Hap1 and greatly enhanced the interaction of Hap1 with Hsp90. These results show that the biological functions of HRMs and Hsp90 are highly sensitive to structural changes. The unique role of HRM7 in heme activation stems from its specific structural environment, not its mere presence. Likewise, the role of Hsp90 in Hap1 activation is dictated by the conformational or structural state of Hap1, not by the mere strength of Hap1-Hsp90 interaction. It appears likely that HRM7 and Hsp90 act together to promote the Hap1 conformational changes that are necessary for Hap1 activation. Such fundamental mechanisms of HRM-Hsp90 cooperation may operate in diverse regulatory systems to mediate signal transduction. PMID- 12897156 TI - Estrogen receptor-dependent proteasomal degradation of the glucocorticoid receptor is coupled to an increase in mdm2 protein expression. AB - Glucocorticoids and estrogens regulate a number of vital physiological processes. We developed a model breast cancer cell line, MCF-7 M, to examine potential mechanisms by which the ligand-bound estrogen receptor (ER) regulates glucocorticoid receptor (GR)-mediated transcription. MCF-7 cells, which endogenously express ERalpha, were stably transfected with mouse mammary tumor virus promoter-luciferase (MMTV-LUC) reporter and GR expression constructs. Our results demonstrate that treatment with estrogen agonists (17beta-estradiol [E2], diethylstilbestrol, genistein), but not antagonists (tamoxifen or raloxifene), for 48 h inhibits GR-mediated MMTV-LUC transcription and chromatin remodeling. Furthermore, estrogen agonists inhibit glucocorticoid induction of p21 mRNA and protein levels, suggesting that the repressive effect applies to other GR regulated genes and proteins in MCF-7 cells. Importantly, GR transcriptional activity is compromised because treatment with estrogen agonists down regulates GR protein levels. The protein synthesis inhibitor cycloheximide and the proteasome inhibitor MG132 block E2-mediated decrease in GR protein levels, suggesting that estrogen agonists down regulate the GR via the proteasomal degradation pathway. In support of this, we demonstrate that E2-mediated GR degradation is coupled to an increase in p53 and its key regulator protein Mdm2 (murine double minute 2), an E3 ubiquitin ligase shown to target the GR for degradation. Using the chromatin immunoprecipitation assay, we demonstrate an E2 dependent recruitment of ERalpha to the Mdm2 promoter, suggesting a role of ER in the regulation of Mdm2 protein expression and hence the enhanced GR degradation in the presence of estrogen agonists. Our study shows that cross talk between the GR and ER involves multiple signaling pathways, indicative of the mechanistic diversity within steroid receptor-regulated transcription. PMID- 12897157 TI - Activation of heat shock genes is not necessary for protection by heat shock transcription factor 1 against cell death due to a single exposure to high temperatures. AB - Heat shock response, which is characterized by the induction of a set of heat shock proteins, is essential for induced thermotolerance and is regulated by heat shock transcription factors (HSFs). Curiously, HSF1 is essential for heat shock response in mammals, whereas in avian HSF3, an avian-specific factor is required for the burst activation of heat shock genes. Amino acid sequences of chicken HSF1 are highly conserved with human HSF1, but those of HSF3 diverge significantly. Here, we demonstrated that chicken HSF1 lost the ability to activate heat shock genes through the amino-terminal domain containing an alanine rich sequence and a DNA-binding domain. Surprisingly, chicken and human HSF1 but not HSF3 possess a novel function that protects against a single exposure to mild heat shock, which is not mediated through the activation of heat shock genes. Overexpression of HSF1 mutants that could not bind to DNA did not restore the susceptibility to cell death in HSF1-null cells, suggesting that the new protective role of HSF1 is mediated through regulation of unknown target genes other than heat shock genes. These results uncover a novel role of vertebrate HSF1, which has been masked under the roles of heat shock proteins. PMID- 12897158 TI - Occupancy and function of the -150 sterol regulatory element and -65 E-box in nutritional regulation of the fatty acid synthase gene in living animals. AB - Upstream regulatory factor (USF) and sterol regulatory element binding protein (SREBP) play key roles in the transcriptional regulation of the fatty acid synthase (FAS) gene by feeding and insulin. Due to the dual binding specificity of SREBP, as well as the presence of multiple consensus sites for these transcription factors in the FAS promoter, their physiologically relevant functional binding sites have been controversial. Here, in order to determine the occupancy of the putative USF and SREBP binding sites, we examined their protein DNA interactions in living animals by using formaldehyde cross-linking and immunoprecipitation of chromatin and tested the function of these elements by employing mice transgenic for a reporter gene driven by various 5' deletions as well as site-specific mutations of the FAS promoter. We show that the -332 and 65 E-boxes are bound by USF in both fasted and refed mice, while the -150 SRE is bound by SREBP-1 only in refed mice. We also found that mutation of either the 150 SRE or the -65 E-box abolishes the feeding-induced activation of the FAS promoter in transgenic mice. Furthermore, in vivo occupancy of the FAS promoter by SREBP in the fed state can be prevented by mutation not only of the -150 SRE but, unexpectedly, of the -65 E-box as well. We conclude that the FAS promoter is activated during refeeding via the induced binding of SREBP to the -150 SRE and that USF binding to the -65 E-box is also required for SREBP binding and activation of the FAS promoter. PMID- 12897159 TI - Polysialic acid directs tumor cell growth by controlling heterophilic neural cell adhesion molecule interactions. AB - Polysialic acid (PSA), a carbohydrate polymer attached to the neural cell adhesion molecule (NCAM), promotes neural plasticity and tumor malignancy, but its mode of action is controversial. Here we establish that PSA controls tumor cell growth and differentiation by interfering with NCAM signaling at cell-cell contacts. Interactions between cells with different PSA and NCAM expression profiles were initiated by enzymatic removal of PSA and by ectopic expression of NCAM or PSA-NCAM. Removal of PSA from the cell surface led to reduced proliferation and activated extracellular signal-regulated kinase (ERK), inducing enhanced survival and neuronal differentiation of neuroblastoma cells. Blocking with an NCAM-specific peptide prevented these effects. Combinatorial transinteraction studies with cells and membranes with different PSA and NCAM phenotypes revealed that heterophilic NCAM binding mimics the cellular responses to PSA removal. In conclusion, our data demonstrate that PSA masks heterophilic NCAM signals, having a direct impact on tumor cell growth. This provides a mechanism for how PSA may promote the genesis and progression of highly aggressive PSA-NCAM-positive tumors. PMID- 12897160 TI - Physical and functional interaction between DNA ligase IIIalpha and poly(ADP Ribose) polymerase 1 in DNA single-strand break repair. AB - The repair of DNA single-strand breaks in mammalian cells is mediated by poly(ADP ribose) polymerase 1 (PARP-1), DNA ligase IIIalpha, and XRCC1. Since these proteins are not found in lower eukaryotes, this DNA repair pathway plays a unique role in maintaining genome stability in more complex organisms. XRCC1 not only forms a stable complex with DNA ligase IIIalpha but also interacts with several other DNA repair factors. Here we have used affinity chromatography to identify proteins that associate with DNA ligase III. PARP-1 binds directly to an N-terminal region of DNA ligase III immediately adjacent to its zinc finger. In further studies, we have shown that DNA ligase III also binds directly to poly(ADP-ribose) and preferentially associates with poly(ADP-ribosyl)ated PARP-1 in vitro and in vivo. Our biochemical studies have revealed that the zinc finger of DNA ligase III increases DNA joining in the presence of either poly(ADP ribosyl)ated PARP-1 or poly(ADP-ribose). This provides a mechanism for the recruitment of the DNA ligase IIIalpha-XRCC1 complex to in vivo DNA single-strand breaks and suggests that the zinc finger of DNA ligase III enables this complex and associated repair factors to locate the strand break in the presence of the negatively charged poly(ADP-ribose) polymer. PMID- 12897161 TI - Support for a meiotic recombination initiation complex: interactions among Rec102p, Rec104p, and Spo11p. AB - Initiation of meiotic recombination in the yeast Saccharomyces cerevisiae requires at least 10 gene products. The initiation event creates double-strand breaks, which are then processed by other recombination enzymes. A variety of classical observations, such as the existence of recombination nodules, have suggested that the proteins catalyzing recombination form a complex. A variety of lines of evidence indicate that Rad50p, Mre11p, and Xrs2p interact, and genetic data suggesting interactions between Rec102p and Rec104p have been reported. It has recently been shown that Spo11p coimmunoprecipitates with Rec102p in meiosis as well. In this paper, we provide genetic and biochemical evidence that the meiosis-specific proteins Rec102p, Rec104p, and Spo11p all interact with each other in meiosis. Furthermore, we demonstrate that the interaction between Rec102p and Spo11p does not require Rec104p. Likewise, the interaction between Rec104p and Rec102p does not require Spo11p, although Spo11p may stabilize that association. The interactions suggest that Spo11p, Rec102p, and Rec104p may form a trimeric complex during the initiation of recombination. PMID- 12897162 TI - Replication checkpoint kinase Cds1 regulates recombinational repair protein Rad60. AB - Genome integrity is protected by Cds1 (Chk2), a checkpoint kinase that stabilizes arrested replication forks. How Cds1 accomplishes this task is unknown. We report that Cds1 interacts with Rad60, a protein required for recombinational repair in fission yeast. Cds1 activation triggers Rad60 phosphorylation and nuclear delocalization. A Rad60 mutant that inhibits regulation by Cds1 renders cells specifically sensitive to replication fork arrest. Genetic and biochemical studies indicate that Rad60 functions codependently with Smc5 and Smc6, subunits of an SMC (structural maintenance of chromosomes) complex required for recombinational repair. These studies indicate that regulation of Rad60 is an important part of the replication checkpoint response controlled by Cds1. We propose that control of Rad60 regulates recombination events at stalled forks. PMID- 12897163 TI - Pex13 inactivation in the mouse disrupts peroxisome biogenesis and leads to a Zellweger syndrome phenotype. AB - Zellweger syndrome is the archetypical peroxisome biogenesis disorder and is characterized by defective import of proteins into the peroxisome, leading to peroxisomal metabolic dysfunction and widespread tissue pathology. In humans, mutations in the PEX13 gene, which encodes a peroxisomal membrane protein necessary for peroxisomal protein import, can lead to a Zellweger phenotype. To develop mouse models for this disorder, we have generated a targeted mouse with a loxP-modified Pex13 gene to enable conditional Cre recombinase-mediated inactivation of Pex13. In the studies reported here, we crossed these mice with transgenic mice that express Cre recombinase in all cells to generate progeny with ubiquitous disruption of Pex13. The mutant pups exhibited many of the clinical features of Zellweger syndrome patients, including intrauterine growth retardation, severe hypotonia, failure to feed, and neonatal death. These animals lacked morphologically intact peroxisomes and showed deficient import of matrix proteins containing either type 1 or type 2 targeting signals. Biochemical analyses of tissue and cultured skin fibroblasts from these animals indicated severe impairment of peroxisomal fatty acid oxidation and plasmalogen synthesis. The brains of these animals showed disordered lamination in the cerebral cortex, consistent with a neuronal migration defect. Thus, Pex13(-/-) mice reproduce many of the features of Zellweger syndrome and PEX13 deficiency in humans. PMID- 12897165 TI - Peripheral chemoreflex and baroreflex interactions in cardiovascular regulation in humans. AB - We tested the hypothesis that activation of peripheral chemoreceptors with acute isocapnic hypoxia resets arterial baroreflex control of both heart rate and sympathetic vasoconstrictor outflow to higher pressures, resulting in increased heart rate and muscle sympathetic nerve activity without changes in baroreflex sensitivity. We further hypothesized that this resetting would not occur during isocapnic hyperpnoea at the same breathing rate and depth as during isocapnic hypoxia. In 12 healthy, non-smoking, normotensive subjects (6 women, 6 men, 19-36 years), we assessed baroreflex control of heart rate and muscle sympathetic nerve activity using the modified Oxford technique during normoxia, isocapnic hyperpnoea, and isocapnic hypoxia (85 % arterial O2 saturation). While isocapnic hyperpnoea did not alter heart rate, arterial pressure, or sympathetic outflow, hypoxia increased heart rate from 61.9 +/- 1.8 to 74.7 +/- 2.7 beats min-1 (P < 0.05), increased mean arterial pressure from 97.4 +/- 2.0 to 103.9 +/- 3.3 mmHg (P < 0.05), and increased sympathetic activity 22 +/- 13 % relative to normoxia and 72 +/- 21 % (P < 0.05) relative to hyperpnoea alone. The sensitivity for baroreflex control of both heart rate and sympathetic activity was not altered by either hypoxia or hyperpnoea. Thus, it appears that acute activation of peripheral chemoreceptors with isocapnic hypoxia resets baroreflex control of both heart rate and sympathetic activity to higher pressures without changes in baroreflex sensitivity. Furthermore, these effects appear largely independent of breathing rate and tidal volume. PMID- 12897166 TI - Dual action of hydrogen peroxide on synaptic transmission at the frog neuromuscular junction. AB - There is evidence that reactive oxygen species (ROS) are produced and released during neuromuscular activity, but their role in synaptic transmission is not known. Using a two-electrode voltage-clamp technique, at frog neuromuscular junctions, the action H2O2 on end-plate currents (EPC) was studied to determine the targets for this membrane-permeable ROS. In curarized or cut muscles, micromolar concentrations of H2O2 increased the amplitude of EPCs. Higher (> 30 microM) doses inhibited EPCs and prolonged current decay. These effects were presynaptic since H2O2 did not change the amplitude or duration of miniature EPCs (although it reduced the rate of spontaneous release at high concentrations). Quantal analysis and deconvolution methods showed that facilitation of EPCs was due to increased quantal release, while depression was accompanied by temporal dispersion of evoked release. Extracellular recordings revealed prolonged presynaptic Ca2+ entry in the presence of high H2O2. Both low and high H2O2 increased presynaptic potentiation during high-frequency stimulation. Pro-oxidant Fe2+ did not affect facilitation by low doses of H2O2 but augmented the inhibition of EPCs by high H2O2, indicating involvement of hydroxyl radicals. High Mg2+ and the ROS scavenger N-acetylcysteine eliminated both the facilitatory and depressant effects of H2O2. The facilitatory effect of H2O2 was prevented by protein kinase C (PKC) inhibitors and 4beta-phorbol 12-myristate, 13-acetate (PMA), an activator of PKC. PKC inhibitors but not PMA also abolished the depressant effect of H2O2. Our data suggest complex presynaptic actions of H2O2, which could serve as a fast feedback modulator of intense neuromuscular transmission. PMID- 12897167 TI - Cold exposure induces tissue-specific modulation of the insulin-signalling pathway in Rattus norvegicus. AB - Cold exposure provides a reproducible model of improved glucose turnover accompanied by reduced steady state and glucose-induced insulin levels. In the present report we performed immunoprecipitation and immunoblot studies to evaluate the initial and intermediate steps of the insulin-signalling pathway in white and brown adipose tissues, liver and skeletal muscle of rats exposed to cold. Basal and glucose-induced insulin secretion were significantly impaired, while glucose clearance rates during a glucose tolerance test and the constant for glucose decay during a 15 min insulin tolerance test were increased, indicating a significantly improved glucose turnover and insulin sensitivity in rats exposed to cold. Evaluation of protein levels and insulin-induced tyrosine (insulin receptor, insulin receptor substrates (IRS)-1 and -2, ERK (extracellular signal-related kinase)) or serine (Akt; protein kinase B) phosphorylation of proteins of the insulin signalling cascade revealed a tissue-specific pattern of regulation of the molecular events triggered by insulin such that in white adipose tissue and skeletal muscle an impaired molecular response to insulin was detected, while in brown adipose tissue an enhanced response to insulin was evident. In muscle and white and brown adipose tissues, increased 2-deoxy-D glucose (2-DG) uptake was detected. Thus, during cold exposure there is a tissue specific regulation of the insulin-signalling pathway, which seems to favour heat producing brown adipose tissue. Nevertheless, muscle and white adipose tissue are able to take up large amounts of glucose, even in the face of an apparent molecular resistance to insulin. PMID- 12897171 TI - Conductance of GABAA channels activated by pentobarbitone in hippocampal neurons from newborn rats. AB - Neurons were obtained from the CA1 region of the hippocampus of newborn rats and maintained in culture. Channels were activated by pentobarbitone in cell attached, inside-out or outside-out patches, normally by applying pentobarbitone in flowing bath solution. Currents were outwardly rectifying and blocked by bicuculline, properties of GABAA channels in these cells. Maximum channel conductance increased as pentobarbitone concentration was increased to 500 microM but conductance then decreased as pentobarbitone concentration was raised further. The best fit of a Hill-type equation to the relationship between maximum channel conductance and pentobarbitone concentration (up to 500 microM) gave an EC50 of 41 microM, a maximum conductance of 36 pS and a Hill coefficient of 1.6. Bicuculline decreased the maximum conductance of the channels activated by pentobarbitone, with an IC50 of 224 microM. Diazepam increased channel conductance, with a maximum effect being obtained with 1 microM diazepam. Diazepam (1 microM) decreased the EC50 of the pentobarbitone effect on channel conductance from 41 microM to 7.2 microM and increased maximum conductance to 72 pS. We conclude that GABAA channel conductance is related to the concentration of the allosteric agonist pentobarbitone. PMID- 12897173 TI - Role of the hyperpolarization-activated cation current (Ih) in pacemaker activity in area postrema neurons of rat brain slices. AB - To clarify the functional properties of the hyperpolarization-activated cation current (Ih) as a pacemaker current in area postrema neurons, whole-cell recordings were made in visually identified cells in rat brain slices. The activation of Ih was identified in approximately 62 % of area postrema neurons tested. The cells displaying Ih showed a depolarizing "sag" in responses to hyperpolarizing current injection in current-clamp mode. The reversal potential for the Ih was -36 mV, and this was shown to depend on the external concentration of Na+ and K+ ions. Extracellular Cs+ ions (2 mM) and ZD7288 (100 microM), a potent selective Ih channel antagonist, blocked Ih and induced a membrane potential hyperpolarization, suggesting the sustained activation of Ih near the resting potential and a contribution from Ih to membrane potential maintenance at more depolarized levels. In contrast, extracellular Ba2+ ions caused a depolarization of the membrane potential, suggesting the blockade of inward rectifier K+ currents. ZD7288 decreased the spontaneous discharge rate by prolonging the slow depolarization between two spikes, with minimal effect on the amplitude of the afterhyperpolarization or action potential waveforms. Ih stabilized the latency of rebound action potentials. Ih was weakly activated by external 8-bromoadenosine 3',5' cyclic monophosphate (1 mM) or forskolin (50-100 microM), indicating that the Ih channel subtypes in area postrema cells could be modulated by intracellular cAMP. Our findings indicate that Ih contributes to the subthreshold membrane and firing properties of rat area postrema neurons and may regulate their resting membrane potential and firing patterns. PMID- 12897175 TI - Protein phosphatases decrease sarcoplasmic reticulum calcium content by stimulating calcium release in cardiac myocytes. AB - Phosphorylation/dephosphorylation of Ca2+ transport proteins by cellular kinases and phosphatases plays an important role in regulation of cardiac excitation contraction coupling; furthermore abnormal protein kinase and phosphatase activities have been implicated in heart failure. However, the precise mechanisms of action of these enzymes on intracellular Ca2+ handling in normal and diseased hearts remains poorly understood. We have investigated the effects of protein phosphatases PP1 and PP2A on spontaneous Ca2+ sparks and SR Ca2+ load in myocytes permeabilized with saponin. Exposure of myocytes to PP1 or PP2A caused a dramatic increase in frequency of Ca2+ sparks followed by a nearly complete disappearance of events. These effects were accompanied by depletion of the SR Ca2+ stores, as determined by application of caffeine. These changes in Ca2+ release and SR Ca2+ load could be prevented by the inhibitors of PP1 and PP2A phosphatase activities okadaic acid and calyculin A. At the single channel level, PP1 increased the open probability of RyRs incorporated into lipid bilayers. PP1-mediated RyR dephosphorylation in our permeabilized myocytes preparations was confirmed biochemically by quantitative immunoblotting using a phosphospecific anti-RyR antibody. Our results suggest that increased intracellular phosphatase activity stimulates RyR-mediated SR Ca2+ release leading to depleted SR Ca2+ stores in cardiac myocytes. PMID- 12897176 TI - Sympathetic nerves inhibit conducted vasodilatation along feed arteries during passive stretch of hamster skeletal muscle. AB - Ascending vasodilatation is integral to blood flow control in exercising skeletal muscle and is attributable to conduction from intramuscular arterioles into proximal feed arteries. Passive stretch of skeletal muscle can impair muscle blood flow but the mechanism is not well understood. We hypothesized that the conduction of vasodilatation along feed arteries can be modulated by changes in muscle length. In anaesthetized hamsters, acetylcholine (ACh) microiontophoresis triggered conducted vasodilatation along feed arteries (diameter, 50-70 microm) of the retractor muscle secured at 100 % resting length or stretched by 30 %. At 100 % length, ACh evoked local dilatation (> 30 microm) and this response conducted rapidly along the feed artery (14 +/- 1 microm dilatation at 1600 microm upstream). During muscle stretch, feed arteries constricted approximately 10 microm (P < 0.05) and local vasodilatation to ACh was maintained while conducted vasodilatation was reduced by half (P < 0.01). Resting diameter and conduction recovered upon restoring 100 % length. Sympathetic nerve stimulation (4-8 Hz) produced vasoconstriction and attenuated conduction in the manner observed during muscle stretch, as did noradrenaline or phenylephrine (10 nM). Inhibiting nitric oxide production (Nomega-nitro-L-arginine, 50 microM) produced similar vasoconstriction yet had no effect on conduction. Phentolamine, prazosin, or tetrodotoxin (1 microM) during muscle stretch abolished vasoconstriction and restored conduction. Inactivation of sensory nerves with capsaicin had no effect on vasomotor responses. Thus, muscle stretch can attenuate conducted vasodilatation by activating alpha-adrenoreceptors on feed arteries through noradrenaline released from perivascular sympathetic nerves. This autonomic feedback mechanism can restrict muscle blood flow during passive stretch. PMID- 12897177 TI - Non-contractile cells with thin processes resembling interstitial cells of Cajal found in the wall of guinea-pig mesenteric arteries. AB - Arterial interstitial cells of Cajal (ICC)-like cells (AIL cells) with a multipolar, irregular, elongated shape and with numerous thin (often less than 1 microm), sometimes branching, processes with lengths up to approximately 60 microm were isolated enzymatically from 1st to 7th order branches of guinea-pig mesenteric artery. Some of the processes of AIL cells were growing (average speed approximately 0.15 microm min-1) and their growth was blocked by 10 microM latrunculin B, an inhibitor of actin polymerisation. Staining with BODIPY phalloidin, a fluorescent dye selective for F-actin, showed the presence of F actin in the processes of AIL cells. Voltage clamp of single AIL cells revealed an inward current that was four times more dense than in myocytes and was abolished by 10 microM nicardipine, and an outward current carried exclusively by potassium ions that was reduced by 1 mM 4-aminopyridine and/or 100 nM iberiotoxin but unaffected by 10 nM dendrotoxin-K. Imaging of intracellular ionised calcium with fluo-4 using a laser scanning confocal microscope showed local or global calcium transients lasting several seconds in approximately 28 % of AIL cells. When membrane current was recorded simultaneously, the calcium transients were found to correspond to long-lasting transient outward currents, which occurred at potentials positive to -40 mV. Unlike myocytes, AIL cells did not contract in response to 1 mM caffeine or 5 microM noradrenaline, although they responded with a [Ca2+]i increase. The segments of intact arteries did not stain for c-kit, a marker of ICCs. Single AIL cells stained positive for vimentin, desmin and smooth muscle myosin. The presence of ICC-like cells is demonstrated for the first time in the media of resistance arteries. PMID- 12897178 TI - Inhibition of nitric oxide synthase by L-NAME speeds phase II pulmonary .VO2 kinetics in the transition to moderate-intensity exercise in man. AB - There is evidence that the rate at which oxygen uptake (.VO2) rises at the transition to higher metabolic rates within the moderate exercise intensity domain is modulated by oxidative enzyme inertia, and also that nitric oxide regulates mitochondrial function through competitive inhibition of cytochrome c oxidase in the electron transport chain. We therefore hypothesised that inhibition of nitric oxide synthase (NOS) by nitro-L-arginine methyl ester (L NAME) would alleviate the inhibition of mitochondrial .VO2 by nitric oxide and result in a speeding of .VO2 kinetics at the onset of moderate-intensity exercise. Seven males performed square-wave transitions from unloaded cycling to a work rate requiring 90 % of predetermined gas exchange threshold with and without prior intravenous infusion of L-NAME (4 mg kg-1 in 50 ml saline over 60 min). Pulmonary gas exchange was measured breath-by-breath and .VO2 kinetics were determined from the averaged response to four exercise bouts performed in each condition using a mono-exponential function following elimination of the phase I response. There were no significant differences between the control and L-NAME conditions for baseline .VO2 (means +/- S.E.M. 797 +/- 32 vs. 794 +/- 29), the duration of phase I (15.4 +/- 0.8 vs. 17.2 +/- 0.6), or the steady-state increment in .VO2 above baseline (1000 +/- 83 vs. 990 +/- 85 ml min-1), respectively. However, the phase II time constant of the .VO2 response was significantly smaller following L-NAME infusion (22.1 +/- 2.4 vs. 17.9 +/- 2.3; P < 0.05). These data indicate that inhibition of NOS by L-NAME results in a significant (19 %) speeding of pulmonary .VO2 kinetics in the transition to moderate-intensity cycle exercise in man. At least part of the intrinsic inertia to oxidative metabolism at the onset of moderate-intensity exercise may result from competitive inhibition of mitochondrial .VO2 by nitric oxide at cytochrome c oxidase, although other mechanisms for the effect of L-NAME on .VO2 kinetics remain to be explored. PMID- 12897181 TI - Milestones in human physiology: Muscle energy metabolism and blood flow during contraction. PMID- 12897180 TI - Synaptogenesis in the CNS: an odyssey from wiring together to firing together. AB - To acquire a better comprehension of nervous system function, it is imperative to understand how synapses are assembled during development and subsequently altered throughout life. Despite recent advances in the fields of neurodevelopment and synaptic plasticity, relatively little is known about the mechanisms that guide synapse formation in the central nervous system (CNS). Although many structural components of the synaptic machinery are pre-assembled prior to the arrival of growth cones at the site of their potential targets, innumerable changes, central to the proper wiring of the brain, must subsequently take place through contact mediated cell-cell communications. Identification of such signalling molecules and a characterization of various events underlying synaptogenesis are pivotal to our understanding of how a brain cell completes its odyssey from "wiring together to firing together". Here we attempt to provide a comprehensive overview that pertains directly to the cellular and molecular mechanisms of selection, formation and refinement of synapses during the development of the CNS in both vertebrates and invertebrates. PMID- 12897182 TI - Resistance training and insulin action in humans: effects of de-training. AB - Aerobic endurance training increases insulin action in skeletal muscle, but the effect of resistance training has not been well described. Controversy exists about whether the effect of resistance training is merely due to an increase in muscle mass. We studied the effect of cessation of resistance training in young, healthy subjects by taking muscle biopsies and measuring insulin-mediated whole body and leg glucose uptake rates after 90 days of heavy resistance training (T) and again after 90 days of de-training (dT). Data on leg glucose uptake were expressed relative to accurate measures of leg muscle mass by MRI scanning. Muscle strength (239 +/- 43 vs. 208 +/- 33 N m), quadriceps area (8463 +/- 453 vs. 7763 +/- 329 mm2) and glycogen content (458 +/- 22 vs. 400 +/- 26 mmol (kg dry weight muscle)(-1)) decreased, while myosin heavy chain isoform IIX increased 4-fold in dT vs. T, respectively (all P < 0.05). GLUT4 mRNA levels and enzyme activities and mRNA levels of glycolytic, lipolytic and glyconeogenic enzymes did not change with de-training. Likewise, capillary density did not change. Whole body glucose uptake decreased 11 % and leg glucose uptake decreased from 75 +/- 11 (T) to 50 +/- 6 (dT) nmol min(-1) (mm muscle)(-2) (P < 0.05) at maximal insulin, the latter decrease being due to decreased arterio-femoral venous glucose extraction. The decrease was mainly due to reduced non-oxidative glucose disposal. We have thus shown that 90 days after the termination of heavy resistance training, insulin-mediated glucose uptake rates per unit of skeletal muscle have decreased significantly. PMID- 12897184 TI - A novel and simple method for quantification of small, dense LDL. AB - A preponderance of small, dense (sd) LDL is strongly associated with the development of coronary heart disease, but the method for the measurement of sd LDL is too laborious for clinical use. We report a simple method for the quantification of sd LDL that is applicable to an autoanalyzer. This method consists of two steps: first, to precipitate the lipoprotein of density (d) <1.044 g/ml using heparin-magnesium; and second, to measure LDL-cholesterol in the supernatant by the homogeneous method or apolipoprotein B (apoB) by an immunoturbidometric assay. The cholesterol and apoB values obtained by the precipitation method (45 +/- 26 and 33 +/- 20 mg/dl, respectively) were similar to those obtained in the lipoprotein (d = 1.044-1.063) separated by ultracentrifugation (42 +/- 22 and 31 +/- 17 mg/dl, respectively), and there was an excellent correlation between the two methods for sd LDL-cholesterol (y = 1.05X + 1, r = 0.88, n = 69) and apoB (y = 1.07X, r = 0.90). Sd LDL values had a significant inverse correlation with LDL size. A high correlation was found between sd LDL-cholesterol and apoB values (r = 0.94). Sd LDL value was related to triglyceride, apoB, and LDL-cholesterol, but not to the buoyant LDL level. These results suggest that this precipitation method is a simple and rapid method for the measurement of sd LDL concentration. PMID- 12897185 TI - Quantification of sphingosine and sphinganine from crude lipid extracts by HPLC electrospray ionization tandem mass spectrometry. AB - Sphingosine (SPH) comprises the backbone of sphingolipids and is known as a second messenger involved in the modulation of cell growth, differentiation, and apoptosis. The currently available methods for the quantification of SPH are, in part, complicated, time-consuming, insensitive, or unselective. Therefore, a fast and convenient methodology for the quantification of SPH and the biosynthetic intermediate sphinganine (SPA) was developed. The method is based on an HPLC separation coupled to electrospray ionization tandem mass spectrometry (MS/MS). Quantitation is achieved by the use of a constant concentration of a non naturally occurring internal standard, 17-carbon chain SPH (C17-SPH), together with a calibration curve established by spiking different concentrations of naturally occurring sphingoid bases. SPH and SPA coeluted with C17-SPH, which allows an accurate correction of the analyte response. Interference of the SPH+2 isotope with SPA quantification was corrected by an experimentally determined factor. The limits of detection were 9 fmol for SPH and 21 fmol for SPA. The overall coefficients of variation were 8% and 13% for SPH and SPA, respectively. The developed HPLC-tandem mass spectrometry methodology, with an analysis time of 3.5 min, simple sample preparation, and automated data analysis, allows high throughput quantification of sphingoid bases from crude lipid extracts and is a valuable tool for studies of cellular sphingolipid metabolism and signaling. PMID- 12897186 TI - Inhibition of cholesterol absorption associated with a PPAR alpha-dependent increase in ABC binding cassette transporter A1 in mice. AB - Dietary supplementation with the peroxisome proliferator-activated receptor alpha (PPAR alpha) ligand WY 14,643 gave rise to a 4- to 5-fold increase in the expression of mRNA for the ATP binding cassette transporter A1 (ABCA1) in the intestine of normal mice. There was no effect in the intestine of PPAR alpha-null mice. Consumption of a high-cholesterol diet also increased intestinal ABCA1 expression. The effects of WY 14,643 and the high-cholesterol diet were not additive. WY 14,643 feeding reduced intestinal absorption of cholesterol in the normal mice, irrespective of the dietary cholesterol concentration, and this resulted in lower diet-derived cholesterol and cholesteryl ester concentrations in plasma and liver. At each concentration of dietary cholesterol, there was a similar significant inverse correlation between intestinal ABCA1 mRNA content and the amount of cholesterol absorbed. The fibrate-induced changes in the intestines of the normal mice were accompanied by an increased concentration of the mRNA encoding the sterol-regulatory element binding protein-1c gene (SREBP-1c), a known target gene for the oxysterol receptor liver X receptor alpha (LXR alpha). There was a correlation between intestinal ABCA1 mRNA and SREBP-1c mRNA contents, but not between SREBP-1c mRNA content and cholesterol absorption. These results suggest that PPAR alpha influences cholesterol absorption through modulating ABCA1 activity in the intestine by a mechanism involving LXR alpha. PMID- 12897187 TI - Transport of cholesterol across a BeWo cell monolayer: implications for net transport of sterol from maternal to fetal circulation. AB - The placental transport of various compounds, such as glucose and fatty acids, has been well studied. However, the transport of cholesterol, a sterol essential for proper fetal development, remains undefined in the placenta. Therefore, the purpose of these studies was to examine the transport of cholesterol across a placental monolayer and its uptake by various cholesterol acceptors. BeWo cells, which originated from a human choriocarcinoma, were grown on transwells for 3 days to form a confluent monolayer. The apical side of the cells was radiolabeled with either free cholesterol or LDL cholesteryl ester. After 24 h, the radiolabel was removed and cholesterol acceptors were added to the basolateral chamber. Cholesterol was found to be taken up by the apical surface of the placental monolayer, transported to the basolateral surface of the cell, and effluxed to fetal human serum, fetal HDL, or phospholipid vesicles, but not to apolipoprotein A-I. In addition, increasing the cellular cholesterol concentration further increased the amount of cholesterol transported to the basolateral acceptors. These are the first studies to demonstrate the movement of cholesterol across a placental cell from the maternal circulation (apical side) to the fetal circulation (basolateral side). PMID- 12897188 TI - FXR-mediated down-regulation of CYP7A1 dominates LXRalpha in long-term cholesterol-fed NZW rabbits. AB - We investigated how cholesterol feeding regulates cholesterol 7alpha-hydroxylase (CYP7A1) via the nuclear receptors farnesoid X receptor (FXR) and liver X receptor alpha (LXRalpha) in New Zealand white rabbits. After 1 day of 2% cholesterol feeding, when the bile acid pool size had not expanded, mRNA levels of the FXR target genes short-heterodimer partner (SHP) and sterol 12alpha hydroxylase (CYP8B) were unchanged, indicating that FXR activation remained constant. In contrast, the mRNA levels of the LXRalpha target genes ATP binding cassette transporter A1 (ABCA1) and cholesteryl ester transfer protein (CETP) increased 5-fold and 2.3-fold, respectively, associated with significant increases in hepatic concentrations of oxysterols. Activity and mRNA levels of CYP7A1 increased 2.4 times and 2.2 times, respectively. After 10 days of cholesterol feeding, the bile acid pool size increased nearly 2-fold. SHP mRNA levels increased 4.1-fold while CYP8B declined 64%. ABCA1 mRNA rose 8-fold and CETP mRNA remained elevated. Activity and mRNA of CYP7A1 decreased 60% and 90%, respectively. Feeding cholesterol for 1 day did not enlarge the ligand pool size or change FXR activation, while LXRalpha was activated highly secondary to increased hepatic oxysterols. As a result, CYP7A1 was up-regulated. After 10 days of cholesterol feeding, the bile acid (FXR ligand) pool size increased, which activated FXR and inhibited CYP7A1 despite continued activation of LXRalpha. Thus, in rabbits, when FXR and LXRalpha are activated simultaneously, the inhibitory effect of FXR overrides the stimulatory effect of LXRalpha to suppress CYP7A1 mRNA expression. PMID- 12897189 TI - Novel putative SREBP and LXR target genes identified by microarray analysis in liver of cholesterol-fed mice. AB - High-cholesterol diets elicit changes in gene expression via such transcription factors as sterol-regulatory element binding proteins (SREBPs) and liver X receptors (LXRs). We used Affymetrix microarrays to identify genes in mouse liver regulated by dietary cholesterol (0.0% vs. 0.5% cholesterol wt/wt). Three independent experiments were performed, and data were analyzed with Affymetrix Microarray Suite and ANOVA statistical software. There were 69 unique Unigene clusters consistently regulated by dietary cholesterol (37 downregulated and 32 upregulated). The array results were confirmed by quantitative RT-PCR (Q-PCR) for seven of nine downregulated genes and five of six upregulated genes. A time course of dietary cholesterol feeding over 1 week revealed different temporal patterns of gene regulation for these confirmed genes. Six downregulated genes were examined in transgenic mice overexpressing truncated nuclear forms of SREBP 1a and SREBP-2, and all were induced in these mice. A second microarray analysis of mice treated with the LXR agonist TO901317 confirmed that 13 of the 32 cholesterol upregulated genes were also LXR-activated. This array result was confirmed by Q-PCR for three of three genes. In summary, these studies identified and confirmed six novel dietary cholesterol-regulated genes, three putative SREBP target genes (calcium/calmodulin-dependent protein kinase 1D, fatty acid binding protein 5, and proprotein convertase subtilisin/kexin 9), and three putative LXR target genes (a disintegrin and metalloprotease domain 11, apoptosis-inhibitory 6, and F-box-only protein 3). PMID- 12897190 TI - Studies on the metabolic fate of n-3 polyunsaturated fatty acids. AB - Several different processes involved in the metabolic fate of docosahexaenoic acid (DHA, C22:6n-3) and its precursor in the biosynthesis route, C24:6n-3, were studied. In cultured skin fibroblasts, the oxidation rate of [1-14C] 24:6n-3 was 2.7 times higher than for [1-14C]22:6n-3, whereas [1-14C]22:6n-3 was incorporated 7 times faster into different lipid classes than was [1-14C]24:6n-3. When determining the peroxisomal acyl-CoA oxidase activity, similar specific activities for C22:6(n-3)-CoA and C24:6(n-3)-CoA were found in mouse kidney peroxisomes. Thioesterase activity was measured for both substrates in mouse kidney peroxisomes as well as mitochondria, and C22:6(n-3)-CoA was hydrolyzed 1.7 times faster than C24:6(n-3)-CoA. These results imply that the preferred metabolic fate of C24:6(n-3)-CoA, after its synthesis in the endoplasmic reticulum (ER), is to move to the peroxisome, where it is beta-oxidized, producing C22:6(n-3)-CoA. This DHA-CoA then preferentially moves back, probably as free fatty acid, to the ER, where it is incorporated into membrane lipids. PMID- 12897191 TI - Comparison of the expression and activity of the lipogenic pathway in human and rat adipose tissue. AB - Lipogenesis is considered less active in human than in rat adipose tissue. This could be explained by different nutritional conditions, namely high-carbohydrate (HCHO) diet in rats and high-fat (HF) diet in humans. Adipose tissue was sampled (postabsorptive state) in rats and humans receiving HCHO or HF diets, ad libitum fed humans, and obese subjects. We measured 1) mRNA concentrations of fatty acid synthase (FAS), acetyl-CoA carboxylase 1 (ACC1), sterol regulatory element binding protein 1c (SREBP-1c), and carbohydrate response element binding protein (ChREBP), 2) SREBP-1c protein, and 3) FAS activity. FAS, ACC1, ChREBP, and SREBP1 c mRNA concentrations were unaffected by diet in humans or in rats. FAS and ACC1 mRNA levels were lower in humans than in rats (P < 0.05). FAS activity was unaffected by diet and was lower in humans (P < 0.05). SREBP-1c mRNA concentrations were similar in rats and humans, but the precursor and mature forms of SREBP-1c protein were less abundant in humans (P < 0.05). ChREBP mRNA concentrations were lower in humans than in rats. In conclusion, the lipogenic capacity of adipose tissue is lower in humans than in rats. This is not related to differences in diet and is probably explained by lower abundance of SREBP-1c protein. A decreased expression of ChREBP could also play a role. PMID- 12897192 TI - Dynamics of membrane lipid domains in neuronal cells differentiated in culture. AB - Treatment with methyl-beta-cyclodextrin (MCD) induced a time- and dose-dependent efflux of cholesterol, sphingolipids, and phosphatidylcholine (PC) from cerebellar neurons differentiated in culture. With a "mild" treatment, the loss of cell lipids induced a deep reorganization of the remaining membrane lipids. In fact, the amount of PC associated with a Triton X-100-insoluble membrane fraction (highly enriched in sphingolipids and cholesterol in nontreated cells) was lowered by the treatment. This suggested a reduction of the lipid domain area. However, the cholesterol and sphingolipid enrichment of this fraction remained substantially unchanged, suggesting the existence of dynamic processes aimed at preserving the segregation of cholesterol and sphingolipids in membrane domains. Under these conditions, the lipid membrane domains retained the ability to sort signaling proteins, such as Lyn and c-Src, but cells displayed deep alterations in their membrane permeability. However, normal membrane permeability was restored by loading cells with cholesterol. When MCD treatment was more stringent, a large loss of cell lipids occurred, and the lipid domains were much less enriched in cholesterol and lost the ability to sort specific proteins. The loss of the integrity and properties of lipid domains was accompanied by severe changes in the membrane permeability, distress, and eventually cell death. PMID- 12897193 TI - Genetic regulation of cholesterol absorption and plasma plant sterol levels: commonalities and differences. AB - The molecular basis of the processes that control two closely related traits, the absorption of cholesterol from the intestines and plasma plant sterol levels, are only partially understood. The discovery that mutations in two novel hemitransporters, ATP binding cassette transporter G5 (ABCG5) and ABCG8, underlie a rare inborn error in plant sterol metabolism, beta-sitosterolemia, represents a major breakthrough in this field. More recently, genetic studies in the mouse that mapped loci in linkage with cholesterol absorption and plasma plant sterol levels and studies in humans that examined the relationship of plasma plant sterol levels to sequence variation in the ABCG5/ABCG8 locus suggested the involvement of other genes. Moreover, studies in beta-sitosterolemic patients, in ABCG5/ABCG8-targeted animals, and on a newly developed cholesterol absorption inhibitor, ezetimibe, suggest commonalities and differences in the regulation of the two traits. This review summarizes the evidence for genetic control of cholesterol absorption and plasma plant sterol levels, presents the evidence for commonalities and differences between the two traits, and discusses recent developments and future perspectives in this field. PMID- 12897194 TI - Sex is a potent modifier of the cardiovascular system. PMID- 12897195 TI - Targeted genetic repair: an emerging approach to genetic therapy. PMID- 12897196 TI - Ribozyme-mediated revision of RNA and DNA. PMID- 12897197 TI - Denying the wolf access to sheep's clothing. AB - Hepatitis delta virus (HDV) is a cause of acute and chronic liver disease for which no effective therapy is currently available. Previous research has demonstrated that prenylation of the large HDV antigen is essential for viral assembly. A new report describes a novel small animal model for HDV replication and demonstrates that prenylation inhibitors are highly effective at clearing viremia and thus have potential relevance for the treatment of chronic delta hepatitis. PMID- 12897198 TI - Amyloid beta and Alzheimer disease therapeutics: the devil may be in the details. AB - Alzheimer disease (AD) is characterized by the progressive accumulation of amyloid beta protein (Abeta) in areas of the brain serving cognitive functions such as memory and language. The first of two separate reports (see the related articles beginning on pages 415 and 440) reveals that intrinsic T cell reactivity to the self-antigen Abeta exists in many humans and increases with age. This finding has implications for the design of Abeta vaccines. The second report demonstrates that a number of FDA- approved nonsteroidal anti-inflammatory drugs are capable of lowering Abeta levels in mice. The work suggests that further testing of the therapeutic utility of these types of compounds for the potential treatment of AD is warranted. PMID- 12897199 TI - Endocannabinoids and the regulation of body fat: the smoke is clearing. AB - Endocannabinoids, endogenous ligands of cannabinoid receptor type 1 (CB1), have emerged as novel and important regulators of energy homeostasis. A report in this issue demonstrates reduced body weight, fat mass, and appetite in CB1-/- mice. Examination of the underlying mechanisms reveals a dual role for endocannabinoids as they affect both appetite and peripheral lipolysis. PMID- 12897200 TI - Renal fibrosis: not just PAI-1 in the sky. AB - A delicate balance exists between ECM synthesis and degradation such that interruption of the corresponding pathways results in increased plasminogen activator inhibitor-1 (PAI-1), pathological matrix accumulation, and glomerulosclerosis. A new study demonstrates that therapy with a mutant PAI-1 increases matrix turnover and reduces glomerulosclerosis by competing with endogenous PAI-1, suggesting therapeutic utility in the treatment of fibrotic renal disease. PMID- 12897201 TI - IL-13 and adenosine: partners in a molecular dance? AB - Inflammation and airway remodeling are two responses readily apparent in asthma and other inflammatory disorders of the airway and lungs. Both adenosine and IL 13 play critical roles in contributing pathways. A new study reveals a previously unrecognized interaction between adenosine and IL-13 that indicates a mutual stimulation that may contribute to the nature and severity of airway inflammation and fibrosis. PMID- 12897202 TI - Adenosine mediates IL-13-induced inflammation and remodeling in the lung and interacts in an IL-13-adenosine amplification pathway. AB - IL-13 is an important mediator of inflammation and remodeling. We hypothesized that adenosine accumulation, alterations in adenosine receptors, and adenosine-IL 13 autoinduction are critical events in IL-13-induced pathologies. To test this, we characterized the effects of IL-13 overexpression on the levels of adenosine, adenosine deaminase (ADA) activity, and adenosine receptors in the murine lung. We also determined whether adenosine induced IL-13 in lungs from ADA-null mice. IL-13 induced an inflammatory and remodeling response that caused respiratory failure and death. During this response, IL-13 caused a progressive increase in adenosine accumulation, inhibited ADA activity and mRNA accumulation, and augmented the expression of the A1, A2B, and A3 but not the A2A adenosine receptors. ADA enzyme therapy diminished the IL-13-induced increase in adenosine, inhibited IL-13-induced inflammation, chemokine elaboration, fibrosis, and alveolar destruction, and prolonged the survival of IL-13-transgenic animals. In addition, IL-13 was strongly induced by adenosine in ADA-null mice. These findings demonstrate that adenosine and adenosine signaling contribute to and influence the severity of IL-13-induced tissue responses. They also demonstrate that IL-13 and adenosine stimulate one another in an amplification pathway that may contribute to the nature, severity, progression, and/or chronicity of IL-13 and/or Th2-mediated disorders. PMID- 12897203 TI - The serum protein alpha 2-Heremans-Schmid glycoprotein/fetuin-A is a systemically acting inhibitor of ectopic calcification. AB - Ectopic calcification is a frequent complication of many degenerative diseases. Here we identify the serum protein alpha2-Heremans-Schmid glycoprotein (Ahsg, also known as fetuin-A) as an important inhibitor of ectopic calcification acting on the systemic level. Ahsg-deficient mice are phenotypically normal, but develop severe calcification of various organs on a mineral and vitamin D-rich diet and on a normal diet when the deficiency is combined with a DBA/2 genetic background. This phenotype is not associated with apparent changes in calcium and phosphate homeostasis, but with a decreased inhibitory activity of the Ahsg-deficient extracellular fluid on mineral formation. The same underlying principle may contribute to many calcifying disorders including calciphylaxis, a syndrome of severe systemic calcification in patients with chronic renal failure. Taken together, our data demonstrate a critical role of Ahsg as an inhibitor of unwanted mineralization and provide a novel therapeutic concept to prevent ectopic calcification accompanying various diseases. PMID- 12897204 TI - Hepatic lipase expression in macrophages contributes to atherosclerosis in apoE deficient and LCAT-transgenic mice. AB - Hepatic lipase (HL) has a well-established role in lipoprotein metabolism. However, its role in atherosclerosis is poorly understood. Here we demonstrate that HL deficiency raises the proatherogenic apoB-containing lipoprotein levels in plasma but reduces atherosclerosis in lecithin cholesterol acyltransferase (LCAT) transgenic (Tg) mice, similar to results previously observed with HL deficient apoE-KO mice. These findings suggest that HL has functions that modify atherogenic risk that are separate from its role in lipoprotein metabolism. We used bone marrow transplantation (BMT) to generate apoE-KO and apoE-KO x HL-KO mice, as well as LCAT-Tg and LCAT-Tg x HL-KO mice, chimeric for macrophage HL gene expression. Using in situ RNA hybridization, we demonstrated localized production of HL by donor macrophages in the artery wall. We found that expression of HL by macrophages enhances early aortic lesion formation in both apoE-KO and LCAT-Tg mice, without changing the plasma lipid profile, lipoprotein lipid composition, or HL and lipoprotein lipase activities. HL does, however, enhance oxidized LDL uptake by peritoneal macrophages. These combined data demonstrate that macrophage-derived HL significantly contributes to early aortic lesion formation in two independent mouse models and identify a novel mechanism, separable from the role of HL in plasma lipoprotein metabolism, by which HL modulates atherogenic risk in vivo. PMID- 12897205 TI - A mutant, noninhibitory plasminogen activator inhibitor type 1 decreases matrix accumulation in experimental glomerulonephritis. AB - In fibrotic renal disease, elevated TGF-beta and angiotensin II lead to increased plasminogen activator inhibitor type 1 (PAI-1). PAI-1 appears to reduce glomerular mesangial matrix turnover by inhibiting plasminogen activators, thereby decreasing plasmin generation and plasmin-mediated matrix degradation. We hypothesized that therapy with a mutant human PAI-1 (PAI-1R) that binds to matrix vitronectin but does not inhibit plasminogen activators, would enhance plasmin generation, increase matrix turnover, and decrease matrix accumulation in experimental glomerulonephritis. Three experimental groups included normal, untreated disease control, and PAI-1R-treated nephritic rats. Plasmin generation by isolated day 3 glomeruli was dramatically decreased by 69%, a decrease that was reversed 43% (P < 0.02) by in vivo PAI-1R treatment. At day 6, animals treated with PAI-1R showed significant reductions in proteinuria (48%, P < 0.02), glomerular staining for periodic acid-Schiff positive material (33%, P < 0.02), collagen I (28%, P < 0.01), collagen III (34%, P < 0.01), fibronectin (48%, P < 0.01), and laminin (41%, P < 0.01), and in collagen I (P < 0.01) and fibronectin mRNA levels (P < 0.02). Treatment did not alter overexpression of TGF-beta1 and PAI-1 mRNAs, although TGF-beta1 protein was significantly reduced. These observations strongly support our hypothesis that PAI-1R reduces glomerulosclerosis by competing with endogenous PAI-1, restoring plasmin generation, inhibiting inflammatory cell infiltration, decreasing local TGF-beta1 concentration, and reducing matrix accumulation. PMID- 12897206 TI - Critical role of galectin-3 in phagocytosis by macrophages. AB - Galectin-3 is a member of a large family of animal lectins. This protein is expressed abundantly by macrophages, but its function in this cell type is not well understood. We have studied the effect of galectin-3 gene targeting on phagocytosis, a major function of macrophages. Compared with wild-type macrophages, galectin-3-deficient (gal3-/-) cells exhibited reduced phagocytosis of IgG-opsonized erythrocytes and apoptotic thymocytes in vitro. In addition, gal3-/- mice showed attenuated phagocytic clearance of apoptotic thymocytes by peritoneal macrophages in vivo. These mice also exhibited reduced IgG-mediated phagocytosis of erythrocytes by Kupffer cells in a murine model of autoimmune hemolytic anemia. Additional experiments indicate that extracellular galectin-3 does not contribute appreciably to the phagocytosis-promoting function of this protein. Confocal microscopic analysis of macrophages containing phagocytosed erythrocytes revealed localization of galectin-3 in phagocytic cups and phagosomes. Furthermore, gal3-/- macrophages exhibited a lower degree of actin rearrangement upon Fcgamma receptor crosslinkage. These results indicate that galectin-3 contributes to macrophage phagocytosis through an intracellular mechanism. Thus, galectin-3 may play an important role in both innate and adaptive immunity by contributing to phagocytic clearance of microorganisms and apoptotic cells. PMID- 12897207 TI - P2Y12 regulates platelet adhesion/activation, thrombus growth, and thrombus stability in injured arteries. AB - The critical role for ADP in arterial thrombogenesis was established by the clinical success of P2Y12 antagonists, currently used at doses that block 40-50% of the P2Y12 on platelets. This study was designed to determine the role of P2Y12 in platelet thrombosis and how its complete absence affects the thrombotic process. P2Y12-null mice were generated by a gene-targeting strategy. Using an in vivo mesenteric artery injury model and real-time continuous analysis of the thrombotic process, we observed that the time for appearance of first thrombus was delayed and that only small, unstable thrombi formed in P2Y12-/- mice without reaching occlusive size, in the absence of aspirin. Platelet adhesion to vWF was impaired in P2Y12-/- platelets. While adhesion to fibrinogen and collagen appeared normal, the platelets in thrombi from P2Y12-/- mice on collagen were less dense and less activated than their WT counterparts. P2Y12-/- platelet activation was also reduced in response to ADP or a PAR-4-activating peptide. Thus, P2Y12 is involved in several key steps of thrombosis: platelet adhesion/activation, thrombus growth, and stability. The data suggest that more aggressive strategies of P2Y12 antagonism will be antithrombotic without the requirement of aspirin cotherapy and may provide benefits even to the aspirin nonresponder population. PMID- 12897208 TI - In vivo antiviral efficacy of prenylation inhibitors against hepatitis delta virus. AB - Hepatitis delta virus (HDV) can dramatically worsen liver disease in patients coinfected with hepatitis B virus (HBV). No effective medical therapy exists for HDV. The HDV envelope requires HBV surface antigen proteins provided by HBV. Once inside a cell, however, HDV can replicate its genome in the absence of any HBV gene products. In vitro, HDV virion assembly is critically dependent on prenyl lipid modification, or prenylation, of its nucleocapsid-like protein large delta antigen. To overcome limitations of current animal models and to test the hypothesis that pharmacologic prenylation inhibition can prevent the production of HDV virions in vivo, we established a convenient mouse-based model of HDV infection capable of yielding viremia. Such mice were then treated with the prenylation inhibitors FTI-277 and FTI-2153. Both agents were highly effective at clearing HDV viremia. As expected, HDV inhibition exhibited duration-of-treatment dependence. These results provide the first preclinical data supporting the in vivo efficacy of prenylation inhibition as a novel antiviral therapy with potential application to HDV and a wide variety of other viruses. PMID- 12897209 TI - Increased T cell reactivity to amyloid beta protein in older humans and patients with Alzheimer disease. AB - Alzheimer disease (AD) is characterized by the progressive deposition of the 42 residue amyloid beta protein (Abeta) in brain regions serving memory and cognition. In animal models of AD, immunization with Abeta results in the clearance of Abeta deposits from the brain. However, a trial of vaccination with synthetic human Abeta1-42 in AD resulted in the development of meningoencephalitis in some patients. We measured cellular immune responses to Abeta in middle-aged and elderly healthy subjects and in patients with AD. A significantly higher proportion of healthy elderly subjects and patients with AD had strong Abeta-reactive T cell responses than occurred in middle-aged adults. The immunodominant Abeta epitopes in humans resided in amino acids 16-33. Epitope mapping enabled the identification of MHC/T cell receptor (TCR) contact residues. The occurrence of intrinsic T cell reactivity to the self-antigen Abeta in humans has implications for the design of Abeta vaccines, may itself be linked to AD susceptibility and course, and appears to be associated with the aging process. PMID- 12897210 TI - The endogenous cannabinoid system affects energy balance via central orexigenic drive and peripheral lipogenesis. AB - The cannabinoid receptor type 1 (CB1) and its endogenous ligands, the endocannabinoids, are involved in the regulation of food intake. Here we show that the lack of CB1 in mice with a disrupted CB1 gene causes hypophagia and leanness. As compared with WT (CB1+/+) littermates, mice lacking CB1 (CB1-/-) exhibited reduced spontaneous caloric intake and, as a consequence of reduced total fat mass, decreased body weight. In young CB1-/- mice, the lean phenotype is predominantly caused by decreased caloric intake, whereas in adult CB1-/- mice, metabolic factors appear to contribute to the lean phenotype. No significant differences between genotypes were detected regarding locomotor activity, body temperature, or energy expenditure. Hypothalamic CB1 mRNA was found to be coexpressed with neuropeptides known to modulate food intake, such as corticotropin-releasing hormone (CRH), cocaine-amphetamine-regulated transcript (CART), melanin-concentrating hormone (MCH), and preproorexin, indicating a possible role for endocannabinoid receptors within central networks governing appetite. CB1-/- mice showed significantly increased CRH mRNA levels in the paraventricular nucleus and reduced CART mRNA levels in the dorsomedial and lateral hypothalamic areas. CB1 was also detected in epidydimal mouse adipocytes, and CB1-specific activation enhanced lipogenesis in primary adipocyte cultures. Our results indicate that the cannabinoid system is an essential endogenous regulator of energy homeostasis via central orexigenic as well as peripheral lipogenic mechanisms and might therefore represent a promising target to treat diseases characterized by impaired energy balance. PMID- 12897212 TI - Griscelli syndrome restricted to hypopigmentation results from a melanophilin defect (GS3) or a MYO5A F-exon deletion (GS1). AB - Griscelli syndrome (GS) is a rare autosomal recessive disorder that associates hypopigmentation, characterized by a silver-gray sheen of the hair and the presence of large clusters of pigment in the hair shaft, and the occurrence of either a primary neurological impairment or a severe immune disorder. Two different genetic forms, GS1 and GS2, respectively, account for the mutually exclusive neurological and immunological phenotypes. Mutations in the gene encoding the molecular motor protein Myosin Va (MyoVa) cause GS1 and the dilute mutant in mice, whereas mutations in the gene encoding the small GTPase Rab27a are responsible for GS2 and the ashen mouse model. We herein present genetic and functional evidence that a third form of GS (GS3), whose expression is restricted to the characteristic hypopigmentation of GS, results from mutation in the gene that encodes melanophilin (Mlph), the ortholog of the gene mutated in leaden mice. We also show that an identical phenotype can result from the deletion of the MYO5A F-exon, an exon with a tissue-restricted expression pattern. This spectrum of GS conditions pinpoints the distinct molecular pathways used by melanocytes, neurons, and immune cells in secretory granule exocytosis, which in part remain to be unraveled. PMID- 12897211 TI - NSAIDs and enantiomers of flurbiprofen target gamma-secretase and lower Abeta 42 in vivo. AB - Epidemiologic studies demonstrate that long-term use of NSAIDs is associated with a reduced risk for the development of Alzheimer disease (AD). In this study, 20 commonly used NSAIDs, dapsone, and enantiomers of flurbiprofen were analyzed for their ability to lower the level of the 42-amino-acid form of amyloid beta protein (Abeta42) in a human H4 cell line. Thirteen of the NSAIDs and the enantiomers of flurbiprofen were then tested in acute dosing studies in amyloid beta protein precursor (APP) transgenic mice, and plasma and brain levels of Abeta and the drug were evaluated. These studies show that (a). eight FDA approved NSAIDs lower Abeta42 in vivo, (b). the ability of an NSAID to lower Abeta42 levels in cell culture is highly predicative of its in vivo activity, (c). in vivo Abeta42 lowering in mice occurs at drug levels achievable in humans, and (d). there is a significant correlation between Abeta42 lowering and levels of ibuprofen. Importantly, flurbiprofen and its enantiomers selectively lower Abeta42 levels in broken cell gamma-secretase assays, indicating that these compounds directly target the gamma-secretase complex that generates Abeta from APP. Of the compounds tested, meclofenamic acid, racemic flurbiprofen, and the purified R and S enantiomers of flurbiprofen lowered Abeta42 levels to the greatest extent. Because R-flurbiprofen reduces Abeta42 levels by targeting gamma secretase and has reduced side effects related to inhibition of cyclooxygenase (COX), it is an excellent candidate for clinical testing as an Abeta42 lowering agent. PMID- 12897213 TI - Abstinence under fire. PMID- 12897214 TI - Recombinant erythropoietin in clinical practice. AB - The introduction of recombinant human erythropoietin (RHuEPO) has revolutionised the treatment of patients with anaemia of chronic renal disease. Clinical studies have demonstrated that RHuEPO is also useful in various non-uraemic conditions including haematological and oncological disorders, prematurity, HIV infection, and perioperative therapies. Besides highlighting both the historical and functional aspects of RHuEPO, this review discusses the applications of RHuEPO in clinical practice and the potential problems of RHuEPO treatment. PMID- 12897215 TI - Osteoarthritis. AB - Osteoarthritis is a chronic degenerative disorder characterised by cartilage loss. It is extremely prevalent in society and is a major cause of disability. It is important to treat osteoarthritis effectively using a multidisciplinary approach tailored to the patient's needs. This paper reviews current thinking on the aetiology, pathogenesis, investigations, and management of osteoarthritis. The paper also discusses the challenges for developing good quality outcome measures for use in large scale multicentre clinical trials for new osteoarthritis treatments, especially disease modifying osteoarthritis drugs. PMID- 12897216 TI - Pulmonary manifestations of sickle cell disease. AB - Pulmonary complications account for significant morbidity and mortality in patients with sickle cell disease. Clinical lung involvement manifests in two major forms: the acute chest syndrome and sickle cell chronic lung disease. Acute chest syndrome is characterised by fever, chest pain, and appearance of a new infiltrate on chest radiograph. Sickle cell chronic lung disease, on the other hand, manifests as radiographic interstitial abnormalities, impaired pulmonary function, and, in its most severe form, by the evidence of pulmonary hypertension. Progress has been made in understanding the pathophysiology and management of these complications. In this review the current knowledge of the mechanism, diagnosis, and treatment of pulmonary complications of sickle cell disease are discussed. PMID- 12897218 TI - Single megadose vitamin A supplementation of Indian mothers and morbidity in breastfed young infants. AB - BACKGROUND AND OBJECTIVE: In developing countries low maternal vitamin A stores combined with increased demands of pregnancy and lactation may lead to its deficiency in breastfed infants. This study evaluates the effects of maternal supplementation with a high dose of vitamin A on the serum retinol levels of exclusively breastfed infants, and their morbidity in the first six months of life. SETTING: Hospital based. STUDY DESIGN: Randomised controlled trial. SUBJECTS AND METHODS: Mothers of the test group (n=150) were orally supplemented with a single dose of retinol (209 micro mol) soon after delivery and were advised exclusive breastfeeding for six months. Before supplementation retinol levels were estimated in the mothers' and newborns' blood, and in colostrum. On follow up, breastmilk and infants' serum retinol contents were assessed monthly for six months. Retinol level <0.7 micro mol/l indicated vitamin A deficiency. Morbidity patterns like vitamin A deficiency, diarrhoea, febrile illnesses, acute respiratory infection, measles, and ear infection were also studied and compared between the two groups. RESULTS: Presupplement mean maternal serum retinol levels were 0.98 and 0.92 micro mol/l and mean breastmilk levels were 3.85 and 3.92 micro mol/l in the test and control groups respectively (p>0.05). Mean cord blood retinol levels were also comparable (0.68 v 0.64 micro mol/l). After supplementation, the test group showed a rise in mean breastmilk retinol content (12.08 v 2.96 micro mol/l) which remained significantly higher for four months. The infants' mean serum retinol level, initially 322.06% of the baseline value, was significantly higher for five months. In the control group, significant numbers of mothers and infants showed deficient breastmilk and serum retinol throughout the follow up (p<0.01). Decreased incidence and duration of various diseases were also found in the test group suggesting lesser morbidity. CONCLUSION: Maternal supplementation with single megadose vitamin A is an effective strategy for vitamin A prophylaxis of exclusively breastfed infants of 0-6 months. PMID- 12897217 TI - Acute and chronic arsenic toxicity. AB - Arsenic toxicity is a global health problem affecting many millions of people. Contamination is caused by arsenic from natural geological sources leaching into aquifers, contaminating drinking water and may also occur from mining and other industrial processes. Arsenic is present as a contaminant in many traditional remedies. Arsenic trioxide is now used to treat acute promyelocytic leukaemia. Absorption occurs predominantly from ingestion from the small intestine, though minimal absorption occurs from skin contact and inhalation. Arsenic exerts its toxicity by inactivating up to 200 enzymes, especially those involved in cellular energy pathways and DNA synthesis and repair. Acute arsenic poisoning is associated initially with nausea, vomiting, abdominal pain, and severe diarrhoea. Encephalopathy and peripheral neuropathy are reported. Chronic arsenic toxicity results in multisystem disease. Arsenic is a well documented human carcinogen affecting numerous organs. There are no evidence based treatment regimens to treat chronic arsenic poisoning but antioxidants have been advocated, though benefit is not proven. The focus of management is to reduce arsenic ingestion from drinking water and there is increasing emphasis on using alternative supplies of water. PMID- 12897219 TI - An informative tattoo. PMID- 12897220 TI - Patients' views on follow up of colorectal cancer: implications for risk communication and decision making. AB - BACKGROUND: Medical views about the clinical value and potential detrimental effect on quality of life of postoperative follow up are divided. There is no literature on the views of British patients with colorectal cancer towards the follow up process. AIM: To investigate patients' views and experiences of follow up of colorectal cancer, and to assess their attitudes towards suggested changes to follow up policy. PATIENTS AND METHODS: A total of 156 asymptomatic and disease-free patients with colorectal cancer were identified from the follow up clinic. Recurrence-free status was confirmed through retrieval of computerised clinic letters. A postal survey using a 39 item piloted questionnaire was undertaken. Data analysis generated descriptive statistics and logistic regression models. RESULTS: A response rate of 61% (95) was obtained. Among these respondents, 63% (60) had undergone initial surgery within three years of the time of the survey, and 86% (82) patients expected a further follow up appointment. Majorities of the sample, ranging from 71% (67) to 96% (91), expressed satisfaction with respect to clinic delays, staff conduct and knowledge about their case, consultation time, and being able to discuss personal problems freely. However some patients reported difficulty in discussing sexual problems at the clinic. Appointment imminence caused anxiety, sleep problems, and decreased appetite in 35% (35), 27% (26), and 8 % (8) of patients respectively. However, 78% (74) patients felt reassured and optimistic for the future after receiving results. Such optimism is not necessarily justified in terms of estimated mortality risks. A majority (78%, 66) stated that they would value finding out about the presence of recurrence even if there would be no survival benefit. Nearly half of the sample (48%, 43) felt that they would disagree with the cessation of follow up in any circumstances. Only 47% (42) and 27% (24) indicated that they would accept follow up by a specialist nurse or their general practitioner, respectively. Attitude to follow up was unrelated to reported anxiety before appointments. Only 22% (19) of the sample could identify risk indicators for recurrence, but 64% (61) agreed that they would like to be told what to look for. DISCUSSION: A sample of patients with colorectal cancer expressed a high degree of satisfaction with hospital follow up. Although a substantial minority reported suffering from pre-visit anxiety, most felt that this disadvantage was compensated for by reassuring results, and believed that investigations did not have a significant negative impact on their quality of life. Respondents valued hospital follow up, and half would reject complete discharge or alternative forms of follow up. These findings demonstrate that patients have a different perception of the risk of recurrence than clinicians who would consider the survival prospects for most patients to be more or less unaffected by follow up interventions. Attempted modifications to follow up policies should be introduced with caution, and should take account of patient understanding of medical reasoning. The findings also raise questions about risk communication with patients. PMID- 12897221 TI - Autonomic neuropathy in patients with hepatic cirrhosis. AB - BACKGROUND: Autonomic neuropathy has been reported in patients with alcoholic liver disease but information on its occurrence in patients with non-alcoholic liver disease is contradictory. AIM: To assess autonomic functions in patients with alcoholic and non-alcoholic liver disease. STUDY DESIGN: Autonomic function using five standard tests was examined in 20 cirrhotics (10 alcoholics and 10 non alcoholics) and 20 age and sex matched controls. The extent of autonomic dysfunction was determined in the patients and a comparison between the characteristics of patients with and without autonomic neuropathy was made. RESULTS: Sixteen (80%) of the cirrhotic subjects were found to have evidence of autonomic neuropathy. Of these, three (15%) patients had early parasympathetic damage, five (25%) had definite parasympathetic damage, and eight (40%) had combined (that is, both parasympathetic and sympathetic) damage. Nine (90%) of the alcoholics and seven (70%) of the non-alcoholics had autonomic dysfunction. Only one patient belonging to the alcoholic group had clinical evidence of peripheral neuropathy. Moreover, there was no significant association between subjective symptoms of autonomic neuropathy and objective evidence of autonomic damage as assessed by autonomic function tests. Autonomic dysfunction was significantly more frequent in advanced liver disease compared with early liver damage. Nine (75%) out of 12 cirrhotic subjects belonging to Child class B and six (85.7%) of the seven patients belonging to Child class C had autonomic neuropathy. CONCLUSION: This study shows that autonomic neuropathy is common in cirrhotic subjects, that it is found with comparable frequency in alcoholics and non-alcoholics, and that it increases in severity with increase in extent of liver damage, suggesting that liver damage contributes to the neurological deficit. PMID- 12897222 TI - Clozapine associated dilated cardiomyopathy. AB - A 31-year-old white man was referred for investigation of a persistent sinus tachycardia. His only significant past medical history was of chronic schizophrenia for which he had been taking clozapine for six years. An electrocardiogram demonstrated sinus tachycardia, voltage criteria for left ventricular hypertrophy, and a prolonged QTc. Echocardiographic findings were consistent with a dilated cardiomyopathy. Serious cardiac complications of clozapine use are rare but have been reported previously. It is important to note that sinus tachycardia may be the only obvious clinical sign, and that complications can manifest months or even years (as in this case) after starting the drug. Patients on clozapine should be informed of potential cardiac symptoms and doctors should maintain a high degree of clinical suspicion throughout the duration of treatment. PMID- 12897223 TI - Metabolic alkalosis and myoclonus. AB - This is the first case reported of vomiting-induced metabolic alkalosis associated with myoclonus. The report describes an unusual presentation of myoclonus secondary to acid-base disturbance caused by recreational drug-induced vomiting. The severe derangement of hyponatraemia, hypokalaemia, and alkalosis appears to have been reasonably well tolerated due to the gradual onset and relatively long history. The causes, mechanism, and management of metabolic alkalosis are discussed. PMID- 12897224 TI - Malignant potential in a Brunner's gland hamartoma. AB - Brunner's gland hamartomas are rare tumours of the duodenum. These lesions have previously been described as being benign, with no malignant potential. A case report is presented of a Brunner's gland hamartoma, whose histology revealed a focus of well marked epithelial dysplasia. This case suggests a dysplastic stage in the natural history of Brunner's gland hamartoma, and questions the malignant potential of these lesions. PMID- 12897225 TI - A gangrenous foot. PMID- 12897226 TI - A bed bound patient. PMID- 12897227 TI - Low HbA1c levels in a poorly controlled diabetic. PMID- 12897228 TI - A misleading swelling of the tongue. PMID- 12897229 TI - Lumbar facet synovial cyst. PMID- 12897230 TI - An unusual cause of a discharging sinus. PMID- 12897236 TI - Thyrotoxic periodic paralysis in western countries. PMID- 12897237 TI - Do I need to be an endocrinologist to diagnose Cushing's disease? PMID- 12897238 TI - Rapid amyloid fiber formation from the fast-folding WW domain FBP28. AB - The WW domains are small proteins that contain a three-stranded, antiparallel beta-sheet. The 40-residue murine FBP28 WW domain rapidly formed twirling ribbon like fibrils at physiological temperature and pH, with morphology typical of amyloid fibrils. These ribbons were unusually wide and well ordered, making them highly suitable for structural studies. Their x-ray and electron-diffraction patterns displayed the characteristic amyloid fiber 0.47-nm reflection of the cross-beta diffraction signature. Both conventional and electron cryomicroscopy showed clearly that the ribbons were composed of many 2.5-nm-wide subfilaments that ran parallel to the long axis of the fiber. There was a region of lower density along the center of each filament. Lateral association of these filaments generated twisted, often interlinked, sheets up to 40 nm wide and many microns in length. The pitch of the helix varied from 60 to 320 nm, depending on the width of the ribbon. The wild-type FBP28 fibers were formed under conditions in which multiexponential folding kinetics is observed in other studies and which was attributed to a change in the mechanism of folding. It is more likely that those phases result from initial events in the off-pathway aggregation observed here. PMID- 12897239 TI - The role of RelMtb-mediated adaptation to stationary phase in long-term persistence of Mycobacterium tuberculosis in mice. AB - Long-term survival of nonreplicating Mycobacterium tuberculosis (Mtb) is ensured by the coordinated shutdown of active metabolism through a broad transcriptional program called the stringent response. In Mtb, this response is initiated by the enzymatic action of RelMtb and deletion of relMtb produces a strain (H37RvDeltarelMtb) severely compromised in the maintenance of long-term viability. Although aerosol inoculation of mice with H37RvDeltarelMtb results in normal initial bacterial growth and containment, the ability of this strain to sustain chronic infection is severely impaired. Significant histopathologic differences were noted in lungs and spleens of mice infected with H37RvDeltarelMtb compared with controls throughout the course of the infection. Microarray analysis revealed that H37RvDeltarelMtb suffers from a generalized alteration of the transcriptional apparatus, as well as specific changes in the expression of virulence factors, cell-wall biosynthetic enzymes, heat shock proteins, and secreted antigens that may alter immune recognition of the recombinant organism. Thus, RelMtb is critical for the successful establishment of persistent infection in mice by altering the expression of antigenic and enzymatic factors that may contribute to successful latent infection. PMID- 12897240 TI - Sex-peptide is the molecular basis of the sperm effect in Drosophila melanogaster. AB - Mating elicits two major changes in the reproductive behavior of many insect females. The egg-laying rate increases and the readiness to accept males (receptivity) is reduced. These postmating responses last approximately 1 week in Drosophila melanogaster. Males that do not transfer sperm but transfer seminal fluid during mating induce a short-term response of 1 day. The long-term response of 1 week requires the presence of sperm (sperm effect). Hence, sperm is essential for the long-term persistence of the postmating responses. Three seminal fluid peptides elicit postmating responses: ovulin, sex-peptide (SP), and DUP99B. Using the technique of targeted mutagenesis by homologous recombination, we have produced males with mutant SP genes. Here, we report that males lacking functional SP elicit only a weak short-term response. However, these males do transfer sperm. Thus, (i) SP is the major agent eliciting the short-term and the long-term postmating responses and (ii) sperm is merely the carrier for SP. The second conclusion is supported by the finding that SP binds to sperm. The 36-aa encoding SP gene is the first small Drosophila gene knocked out with the method of homologous recombination. PMID- 12897241 TI - Melanin-concentrating hormone is a critical mediator of the leptin-deficient phenotype. AB - Energy homeostasis is regulated by a complex network involving peripheral and central signals that determine food intake and energy expenditure. Melanin concentrating hormone (MCH) plays an essential role in this process. Animals treated with MCH develop hyperphagia and obesity. Ablation of the prepro-MCH gene leads to a lean phenotype, as does ablation of the rodent MCH receptor, MCHR-1. MCH is overexpressed in the leptin-deficient ob/ob mouse, and we hypothesized that ablation of MCH in this animal would lead to attenuation of its obese phenotype. Compared with ob/ob animals, mice lacking both leptin and MCH (double null) had a dramatic reduction in body fat. Surprisingly, the hyperphagia of the ob/ob mouse was unaffected. Instead, leanness was secondary to a marked increase in energy expenditure resulting from both increased resting energy expenditure and locomotor activity. Furthermore, double-null mice showed improvements in other parameters impaired in ob/ob mice. Compared with ob/ob mice, double-null animals had increased basal body temperature, improved response to cold exposure, lower plasma glucocorticoid levels, improved glucose tolerance, and reduced expression of stearoyl-CoA desaturase 1 (SCD-1). These results highlight the importance of MCH in integration of energy homeostasis downstream of leptin and, in particular, the role of MCH in regulation of energy expenditure. PMID- 12897242 TI - Defective mammary gland morphogenesis in mice lacking the progesterone receptor B isoform. AB - Progesterone (P) regulates female reproduction via two nuclear receptors, PR-A and PR-B. Although both receptors display overlapping and distinct transcription regulatory properties, their individual physiological roles are unclear. To address the physiological role of PR-A, we generated a mouse model in which expression of PR-B was specifically ablated (PRBKO-/-). We show that selective activation of PR-A in PRBKO-/- mice is sufficient to elicit normal ovarian and uterine responses to P but results in reduced mammary gland morphogenesis. In the absence of PR-B, pregnancy-associated ductal sidebranching and lobuloalveolar development are markedly reduced due to decreased ductal and alveolar epithelial cell proliferation and decreased survival of alveolar epithelium. In an effort to elucidate the molecular genetic signaling pathways that are differentially regulated by PRs in the mammary gland, we have identified receptor activator of nuclear factor kappa B ligand (RANKL) as a paracrine mediator of P-dependent alveologenesis. Further, we demonstrate that the defects in PRBKO-/- mice are associated with an inability of PR-A to activate the RANKL signaling pathway in response to P. Our data indicate that functional interaction between PR-A and PR B is not required for reproductive activity and that selective modulation of PR-A activity by progestin agonists may have a protective effect against both uterine and mammary gland hyperplasias. PMID- 12897243 TI - Morphogenesis of the telencephalic commissure requires scaffold protein JNK interacting protein 3 (JIP3). AB - The murine JNK-interacting protein 3 (JIP3) protein (also known as JSAP1) is expressed exclusively in neurons and has been identified as a scaffold protein for the c-Jun NH2-terminal kinase (JNK) signaling pathway and as an adapter protein for cargo transport by the microtubule motor protein kinesin. To investigate the physiological function of JIP3, we examined the effect of Jip3 gene disruption in mice. The Jip3-/- mice were unable to breathe and died shortly after birth. Microscopic analysis demonstrated that Jip3 gene disruption causes severe defects in the morphogenesis of the telencephalon. Jip3-/- mice lack the telencephalic commissure, a major connection between the left and right hemispheres of the brain. The central nervous system abnormalities of Jip3-/- mice may be accounted for in part by a reduction in signal transduction by RhoA and its effector ROCK. PMID- 12897244 TI - The phosphatidylinositol (PI)-5-phosphate 4-kinase type II enzyme controls insulin signaling by regulating PI-3,4,5-trisphosphate degradation. AB - Phosphatidylinositol-5-phosphate (PI-5-P) is a newly identified phosphoinositide with characteristics of a signaling lipid but no known cellular function. PI-5-P levels are controlled by the type II PI-5-P 4-kinases (PIP4K IIs), a family of kinases that converts PI-5-P into phosphatidylinositol-4,5-bisphosphate (PI-4,5 P2). The PI-5-P pathway is an alternative route for PI-4,5-P2 synthesis as the bulk of this lipid is generated by the canonical pathway in which phosphatidylinositol-4-phosphate (PI-4-P) is the intermediate. Here we examined the effect of activation of the PI-5-P pathway on phosphoinositide 3-kinase (PI3K) signaling by expressing PIP4K II beta in cells that lack this enzyme. Although PIP4K II generates PI-4,5-P2, a substrate for PI3K, expression of this enzyme reduced rather than increased phosphatidylinositol-3,4,5-trisphosphate (PI 3,4,5-P3) levels in cells stimulated with insulin or cells expressing activated PI3K. This reduction in PI-3,4,5-P3 levels resulted in decreased activation of the downstream protein kinase, Akt/PKB. Consistent with these results, expression of IpgD, a bacterial phosphatase that converts PI-4,5-P2 to PI-5-P, resulted in Akt activation, and this effect was partially reversed by PIP4K II beta. PIP4K II beta expression did not impair insulin-dependent association of PI3K with insulin receptor substrate 1 (IRS1) but abbreviated Akt activation, indicating that PIP4K II regulates PI-3,4,5-P3 degradation rather than synthesis. These data support a model in which the PI-5-P pathway controls insulin signaling that leads to Akt activation by regulating a PI-3,4,5-P3 phosphatase. PMID- 12897245 TI - Activation tagging in tomato identifies a transcriptional regulator of anthocyanin biosynthesis, modification, and transport. AB - We have developed a high-throughput T-DNA insertional mutagenesis program in tomato using activation tagging to identify genes that regulate metabolic pathways. One of the activation-tagged insertion lines (ant1) showed intense purple pigmentation from the very early stage of shoot formation in culture, reflecting activation of the biosynthetic pathway leading to anthocyanin accumulation. The purple coloration resulted from the overexpression of a gene that encodes a MYB transcription factor. Vegetative tissues of ant1 plants displayed intense purple color, and the fruit showed purple spotting on the epidermis and pericarp. The gene-to-trait relationship of ant1 was confirmed by the overexpression of ANT1 in transgenic tomato and in tobacco under the control of a constitutive promoter. Suppression subtractive hybridization and RNA hybridization analysis of the purple tomato plants indicated that the overexpression of ANT1 caused the upregulation of genes that encode proteins in both the early and later steps of anthocyanidin biosynthesis as well as genes involved in the glycosylation and transport of anthocyanins into the vacuole. PMID- 12897246 TI - Structural basis for substrate recognition in the salicylic acid carboxyl methyltransferase family. AB - Recently, a novel family of methyltransferases was identified in plants. Some members of this newly discovered and recently characterized methyltransferase family catalyze the formation of small-molecule methyl esters using S-adenosyl-L Met (SAM) as a methyl donor and carboxylic acid-bearing substrates as methyl acceptors. These enzymes include SAMT (SAM:salicylic acid carboxyl methyltransferase), BAMT (SAM:benzoic acid carboxyl methyltransferase), and JMT (SAM:jasmonic acid carboxyl methyltransferase). Moreover, other members of this family of plant methyltransferases have been found to catalyze the N-methylation of caffeine precursors. The 3.0-A crystal structure of Clarkia breweri SAMT in complex with the substrate salicylic acid and the demethylated product S-adenosyl L-homocysteine reveals a protein structure that possesses a helical active site capping domain and a unique dimerization interface. In addition, the chemical determinants responsible for the selection of salicylic acid demonstrate the structural basis for facile variations of substrate selectivity among functionally characterized plant carboxyl-directed and nitrogen-directed methyltransferases and a growing set of related proteins that have yet to be examined biochemically. Using the three-dimensional structure of SAMT as a guide, we examined the substrate specificity of SAMT by site-directed mutagenesis and activity assays against 12 carboxyl-containing small molecules. Moreover, the utility of structural information for the functional characterization of this large family of plant methyltransferases was demonstrated by the discovery of an Arabidopsis methyltransferase that is specific for the carboxyl-bearing phytohormone indole-3-acetic acid. PMID- 12897247 TI - The interaction of two homeobox genes, BREVIPEDICELLUS and PENNYWISE, regulates internode patterning in the Arabidopsis inflorescence. AB - Plant architecture results from the activity of the shoot apical meristem, which initiates leaves, internodes, and axillary meristems. KNOTTED1-like homeobox (KNOX) genes are expressed in specific patterns in the shoot apical meristem and play important roles in plant architecture. KNOX proteins interact with BEL1-like (BELL) homeodomain proteins and together bind a target sequence with high affinity. We have obtained a mutation in one of the Arabidopsis BELL genes, PENNYWISE (PNY), that appears phenotypically similar to the KNOX mutant brevipedicellus (bp). Both bp and pny have randomly shorter internodes and display a slight increase in the number of axillary branches. The double mutant shows a synergistic phenotype of extremely short internodes interspersed with long internodes and increased branching. PNY is expressed in inflorescence and floral meristems and overlaps with BP in a discrete domain of the inflorescence meristem where we propose the internode is patterned. The physical association of the PNY and BP proteins suggests that they participate in a complex that regulates early patterning events in the inflorescence meristem. PMID- 12897248 TI - The MSP1 gene is necessary to restrict the number of cells entering into male and female sporogenesis and to initiate anther wall formation in rice. AB - The function of the novel gene MSP1 (MULTIPLE SPOROCYTE), which controls early sporogenic development, was elucidated by characterizing a retrotransposon-tagged mutation of rice. The MSP1 gene encoded a Leu-rich repeat receptor-like protein kinase. The msp1 mutation gave rise to an excessive number of both male and female sporocytes. In addition, the formation of anther wall layers was disordered and the tapetum layer was lost completely. Although the mutation never affected homologous chromosome pairing and chiasma maintenance, the development of pollen mother cells was arrested at various stages of meiotic prophase I, which resulted in complete male sterility. Meanwhile, plural megaspore mother cells in a mutant ovule generated several megaspores, underwent gametogenesis, and produced germinable seeds when fertilized with wild-type pollen despite disorganized female gametophytes. In situ expression of MSP1 was detected in surrounding cells of male and female sporocytes and some flower tissues, but never in the sporocytes themselves. These results suggest that the MSP1 product plays crucial roles in restricting the number of cells entering into male and female sporogenesis and in initiating anther wall formation in rice. PMID- 12897249 TI - Control of cellulose synthase complex localization in developing xylem. AB - Cellulose synthesis in the developing xylem vessels of Arabidopsis requires three members of the cellulose synthase (CesA) gene family. In young vessels, these three proteins localize within the cell, whereas in older vessels, all three CesA proteins colocalize with bands of cortical microtubules that mark the sites of secondary cell wall deposition. In the absence of one subunit, however, the remaining two subunits are retained in the cell, demonstrating that all three CesA proteins are required to assemble a functional complex. CesA proteins with altered catalytic activity localize normally, suggesting that cellulose synthase activity is not required for this localization. Cortical microtubule arrays are required continually to maintain normal CesA protein localization. By contrast, actin microfilaments do not colocalize with the CesA proteins and are unlikely to play a direct role in their localization. Green fluorescent protein-tagged CesA reveals a novel process in which the structure and/or local environment of the cellulose synthase complex is altered rapidly. PMID- 12897250 TI - The Arabidopsis basic/helix-loop-helix transcription factor family. AB - The basic/helix-loop-helix (bHLH) proteins are a superfamily of transcription factors that bind as dimers to specific DNA target sites and that have been well characterized in nonplant eukaryotes as important regulatory components in diverse biological processes. Based on evidence that the bHLH protein PIF3 is a direct phytochrome reaction partner in the photoreceptor's signaling network, we have undertaken a comprehensive computational analysis of the Arabidopsis genome sequence databases to define the scope and features of the bHLH family. Using a set of criteria derived from a previously defined consensus motif, we identified 147 bHLH protein-encoding genes, making this one of the largest transcription factor families in Arabidopsis. Phylogenetic analysis of the bHLH domain sequences permits classification of these genes into 21 subfamilies. The evolutionary and potential functional relationships implied by this analysis are supported by other criteria, including the chromosomal distribution of these genes relative to duplicated genome segments, the conservation of variant exon/intron structural patterns, and the predicted DNA binding activities within subfamilies. Considerable diversity in DNA binding site specificity among family members is predicted, and marked divergence in protein sequence outside of the conserved bHLH domain is observed. Together with the established propensity of bHLH factors to engage in varying degrees of homodimerization and heterodimerization, these observations suggest that the Arabidopsis bHLH proteins have the potential to participate in an extensive set of combinatorial interactions, endowing them with the capacity to be involved in the regulation of a multiplicity of transcriptional programs. We provide evidence from yeast two hybrid and in vitro binding assays that two related phytochrome-interacting members in the Arabidopsis family, PIF3 and PIF4, can form both homodimers and heterodimers and that all three dimeric configurations can bind specifically to the G-box DNA sequence motif CACGTG. These data are consistent, in principle, with the operation of this combinatorial mechanism in Arabidopsis. PMID- 12897252 TI - An Arabidopsis mutant resistant to thaxtomin A, a cellulose synthesis inhibitor from Streptomyces species. AB - Thaxtomin A is a phytotoxin produced by Streptomyces scabies and other Streptomyces species, the causative agents of common scab disease in potato and other taproot crops. At nanomolar concentrations, thaxtomin causes dramatic cell swelling, reduced seedling growth, and inhibition of cellulose synthesis in Arabidopsis. We identified a mutant of Arabidopsis, designated txr1, that exhibits increased resistance to thaxtomin as a result of a decrease in the rate of toxin uptake. The TXR1 gene was identified by map-based cloning and found to encode a novel, small protein with no apparent motifs or organelle-targeting signals. The protein, which has homologs in all fully sequenced eukaryotic genomes, is expressed in all tissues and during all developmental stages analyzed. Microarray transcript profiling of some 14,300 genes revealed two stomatin-like genes that were expressed differentially in the txr1 mutant and the wild type. We propose that TXR1 is a regulator of a transport mechanism. PMID- 12897254 TI - Altered nuclear distribution of recombination protein RAD51 in maize mutants suggests the involvement of RAD51 in meiotic homology recognition. AB - The recombination protein RAD51 is a component of the meiotic recombination pathway and has been proposed to play a role in the homology search, a process by which homologous chromosomes find each other before they pair in the prophase of meiosis. To study the relationship between recombination and chromosome pairing, we examined the distribution of RAD51 foci on meiotic chromosomes in maize mutants with defects in chromosome pairing. The patterns of RAD51 distribution showed dramatic variation among the meiotic mutants. The mutants generally exhibited significant decreases in the number of RAD51 foci at zygotene, corresponding to the degree of their pairing defects. These results provide evidence for a key role of RAD51 structures in the homology search. PMID- 12897255 TI - Cytoplasmic N-terminal protein acetylation is required for efficient photosynthesis in Arabidopsis. AB - The Arabidopsis atmak3-1 mutant was identified on the basis of a decreased effective quantum yield of photosystem II. In atmak3-1, the synthesis of the plastome-encoded photosystem II core proteins D1 and CP47 is affected, resulting in a decrease in the abundance of thylakoid multiprotein complexes. DNA array based mRNA analysis indicated that extraplastid functions also are altered. The mutation responsible was localized to AtMAK3, which encodes a homolog of the yeast protein Mak3p. In yeast, Mak3p, together with Mak10p and Mak31p, forms the N-terminal acetyltransferase complex C (NatC). The cytoplasmic AtMAK3 protein can functionally replace Mak3p, Mak10p, and Mak31p in acetylating N termini of endogenous proteins and the L-A virus Gag protein. This result, together with the finding that knockout of the Arabidopsis MAK10 homolog does not result in obvious physiological effects, indicates that AtMAK3 function does not require NatC complex formation, as it does in yeast. We suggest that N-acetylation of certain chloroplast precursor protein(s) is necessary for the efficient accumulation of the mature protein(s) in chloroplasts. PMID- 12897251 TI - Target site specificity of the Tos17 retrotransposon shows a preference for insertion within genes and against insertion in retrotransposon-rich regions of the genome. AB - Because retrotransposons are the major component of plant genomes, analysis of the target site selection of retrotransposons is important for understanding the structure and evolution of plant genomes. Here, we examined the target site specificity of the rice retrotransposon Tos17, which can be activated by tissue culture. We have produced 47,196 Tos17-induced insertion mutants of rice. This mutant population carries approximately 500,000 insertions. We analyzed >42,000 flanking sequences of newly transposed Tos17 copies from 4316 mutant lines. More than 20,000 unique loci were assigned on the rice genomic sequence. Analysis of these sequences showed that insertion events are three times more frequent in genic regions than in intergenic regions. Consistent with this result, Tos17 was shown to prefer gene-dense regions over centromeric heterochromatin regions. Analysis of insertion target sequences revealed a palindromic consensus sequence, ANGTT-TSD-AACNT, flanking the 5-bp target site duplication. Although insertion targets are distributed throughout the chromosomes, they tend to cluster, and 76% of the clusters are located in genic regions. The mechanisms of target site selection by Tos17, the utility of the mutant lines, and the knockout gene database are discussed. --The nucleotide sequence data were uploaded to the DDBJ, EMBL, and GenBank nucleotide sequence databases under accession numbers AG020727 to AG025611 and AG205093 to AG215049. PMID- 12897253 TI - Contrasting effects of selection on sequence diversity and linkage disequilibrium at two phytoene synthase loci. AB - We investigated the effects of human selection for yellow endosperm color, representing increased carotenoid content, on two maize genes, the Y1 phytoene synthase and PSY2, a putative second phytoene synthase. Multiple polymorphic sites were identified at Y1 and PSY2 in 75 white and yellow maize inbred lines. Many polymorphic sites showed strong association with the endosperm color phenotype at Y1, but no detectable association was found at PSY2. Nucleotide diversity was equivalent for whites and yellows at PSY2 but was 19-fold less in yellows than in whites at Y1, consistent with the white ancestral state of the gene. The strong sequence haplotype conservation within yellows at Y1 and a significant, negative Tajima's D both verified positive selection for yellow endosperm. We propose that two independent gain-of-function events associated with insertions into the promoter of the Y1 gene and upregulation of expression in endosperm have been incorporated into yellow maize. PMID- 12897256 TI - CBL1, a calcium sensor that differentially regulates salt, drought, and cold responses in Arabidopsis. AB - Although calcium is a critical component in the signal transduction pathways that lead to stress gene expression in higher plants, little is known about the molecular mechanism underlying calcium function. It is believed that cellular calcium changes are perceived by sensor molecules, including calcium binding proteins. The calcineurin B-like (CBL) protein family represents a unique group of calcium sensors in plants. A member of the family, CBL1, is highly inducible by multiple stress signals, implicating CBL1 in stress response pathways. When the CBL1 protein level was increased in transgenic Arabidopsis plants, it altered the stress response pathways in these plants. Although drought-induced gene expression was enhanced, gene induction by cold was inhibited. In addition, CBL1 overexpressing plants showed enhanced tolerance to salt and drought but reduced tolerance to freezing. By contrast, cbl1 null mutant plants showed enhanced cold induction and reduced drought induction of stress genes. The mutant plants displayed less tolerance to salt and drought but enhanced tolerance to freezing. These studies suggest that CBL1 functions as a positive regulator of salt and drought responses and a negative regulator of cold response in plants. PMID- 12897257 TI - Salicylic acid and NPR1 induce the recruitment of trans-activating TGA factors to a defense gene promoter in Arabidopsis. AB - Efforts to elucidate the contributions by transcription factors to plant gene expression will require increasing knowledge of their specific in vivo regulatory associations. We are systematically investigating the role of individual TGA factors in the transcriptional control of pathogenesis-related (PR) defense genes, whose expression is stimulated in leaves by salicylic acid (SA) through a stimulus pathway involving NPR1. We focused on PR-1 because its SA-induced expression in Arabidopsis is mediated by an as-1-type promoter cis element (LS7) that is recognized in vitro by TGA factors. We found that two NPR1-interacting TGA factors, TGA2 and TGA3, are the principal contributors to an LS7 binding activity of leaves that is enhanced by SA through NPR1. The relevance of these findings to PR-1 expression was investigated by the use of chromatin immunoprecipitation, which demonstrated that in vivo these TGA factors are strongly recruited in an SA- and NPR1-dependent manner to the LS7-containing PR-1 promoter. Significantly, the timing of promoter occupancy by these factors is linked to the SA-induced onset and sustained expression of PR-1. Because leaf transfection assays indicate that TGA3 activates transcription, as noted previously for TGA2, these two TGA factors are predicted to make positive contributions to the expression of this target gene. Thus, the findings presented here distinguish among different modes of regulation by these transcription factors and provide strong support for their direct role in the stimulus activated expression of an endogenous defense gene. PMID- 12897258 TI - The Arabidopsis ppi1 mutant is specifically defective in the expression, chloroplast import, and accumulation of photosynthetic proteins. AB - The import of nucleus-encoded proteins into chloroplasts is mediated by translocon complexes in the envelope membranes. A component of the translocon in the outer envelope membrane, Toc34, is encoded in Arabidopsis by two homologous genes, atTOC33 and atTOC34. Whereas atTOC34 displays relatively uniform expression throughout development, atTOC33 is strongly upregulated in rapidly growing, photosynthetic tissues. To understand the reason for the existence of these two related genes, we characterized the atTOC33 knockout mutant ppi1. Immunoblotting and proteomics revealed that components of the photosynthetic apparatus are deficient in ppi1 chloroplasts and that nonphotosynthetic chloroplast proteins are unchanged or enriched slightly. Furthermore, DNA array analysis of 3292 transcripts revealed that photosynthetic genes are moderately, but specifically, downregulated in ppi1. Proteome differences in ppi1 could be correlated with protein import rates: ppi1 chloroplasts imported the ribulose-1,5 bisphosphate carboxylase/oxygenase small subunit and 33-kD oxygen-evolving complex precursors at significantly reduced rates, but the import of a 50S ribosomal subunit precursor was largely unaffected. The ppi1 import defect occurred at the level of preprotein binding, which is consistent with a role for atToc33 during preprotein recognition. The data suggest that atToc33 is involved preferentially in the import of photosynthetic proteins and, by extension, that atToc34 is involved in the import of nonphotosynthetic chloroplast proteins. PMID- 12897259 TI - Arabidopsis AtGPAT1, a member of the membrane-bound glycerol-3-phosphate acyltransferase gene family, is essential for tapetum differentiation and male fertility. AB - Membrane-bound glycerol-3-phosphate acyltransferase (GPAT; EC 2.3.1.15) mediates the initial step of glycerolipid biosynthesis in the extraplastidic compartments of plant cells. Here, we report the molecular characterization of a novel GPAT gene family from Arabidopsis, designated AtGPAT. The corresponding polypeptides possess transmembrane domains and GPAT activity when expressed heterologously in a yeast lipid mutant. The functional significance of one isoform, AtGPAT1, is the focus of the present study. Disruption of the AtGPAT1 gene causes a massive pollen development arrest, and subsequent introduction of the gene into the mutant plant rescues the phenotype, illustrating a pivotal role for AtGPAT1 in pollen development. Microscopic examinations revealed that the gene lesion results in a perturbed degeneration of the tapetum, which is associated with altered endoplasmic reticulum profiles and reduced secretion. In addition to the sporophytic effect, AtGPAT1 also exerts a gametophytic effect on pollen performance, as the competitive ability of a pollen grain to pollinate is dependent on the presence of an AtGPAT1 gene. Deficiency in AtGPAT1 correlates with several fatty acid composition changes in flower tissues and seeds. Unexpectedly, however, a loss of AtGPAT1 causes no significant change in seed oil content. PMID- 12897260 TI - The mitochondrial genome of Chara vulgaris: insights into the mitochondrial DNA architecture of the last common ancestor of green algae and land plants. AB - Mitochondrial DNA (mtDNA) has undergone radical changes during the evolution of green plants, yet little is known about the dynamics of mtDNA evolution in this phylum. Land plant mtDNAs differ from the few green algal mtDNAs that have been analyzed to date by their expanded size, long spacers, and diversity of introns. We have determined the mtDNA sequence of Chara vulgaris (Charophyceae), a green alga belonging to the charophycean order (Charales) that is thought to be the most closely related alga to land plants. This 67,737-bp mtDNA sequence, displaying 68 conserved genes and 27 introns, was compared with those of three angiosperms, the bryophyte Marchantia polymorpha, the charophycean alga Chaetosphaeridium globosum (Coleochaetales), and the green alga Mesostigma viride. Despite important differences in size and intron composition, Chara mtDNA strikingly resembles Marchantia mtDNA; for instance, all except 9 of 68 conserved genes lie within blocks of colinear sequences. Overall, our genome comparisons and phylogenetic analyses provide unequivocal support for a sister-group relationship between the Charales and the land plants. Only four introns in land plant mtDNAs appear to have been inherited vertically from a charalean algar ancestor. We infer that the common ancestor of green algae and land plants harbored a tightly packed, gene-rich, and relatively intron-poor mitochondrial genome. The group II introns in this ancestral genome appear to have spread to new mtDNA sites during the evolution of bryophytes and charalean green algae, accounting for part of the intron diversity found in Chara and land plant mitochondria. PMID- 12897261 TI - Regulation and processing of maize histone deacetylase Hda1 by limited proteolysis. AB - A maize histone deacetylase gene was identified as a homolog of yeast Hda1. The predicted protein corresponds to a previously purified maize deacetylase that is active as a protein monomer with a molecular weight of 48,000 and is expressed in all tissues of germinating embryos. Hda1 is synthesized as an enzymatically inactive protein with an apparent molecular weight of 84,000 that is processed to the active 48-kD form by proteolytic removal of the C-terminal part, presumably via a 65-kD intermediate. The enzymatically inactive 84-kD protein also is part of a 300-kD protein complex of unknown function. The proteolytic cleavage of ZmHda1 is regulated during maize embryo germination in vivo. Expression of the recombinant full-length protein and the 48-kD form confirmed that only the smaller enzyme form is active as a histone deacetylase. In line with this finding, we show that the 48-kD protein is able to repress transcription efficiently in a reporter gene assay, whereas the full-length protein, including the C-terminal part, lacks full repression activity. This report on the processing of Hda1-p84 to enzymatically active Hda1-p48 demonstrates that proteolytic cleavage is a mechanism to regulate the function of Rpd3/Hda1-type histone deacetylases. PMID- 12897262 TI - ARC6 is a J-domain plastid division protein and an evolutionary descendant of the cyanobacterial cell division protein Ftn2. AB - Replication of chloroplasts is essential for achieving and maintaining optimal plastid numbers in plant cells. The plastid division machinery contains components of both endosymbiotic and host cell origin, but little is known about the regulation and molecular mechanisms that govern the division process. The Arabidopsis mutant arc6 is defective in plastid division, and its leaf mesophyll cells contain only one or two grossly enlarged chloroplasts. We show here that arc6 chloroplasts also exhibit abnormal localization of the key plastid division proteins FtsZ1 and FtsZ2. Whereas in wild-type plants, the FtsZ proteins assemble into a ring at the plastid division site, chloroplasts in the arc6 mutant contain numerous short, disorganized FtsZ filament fragments. We identified the mutation in arc6 and show that the ARC6 gene encodes a chloroplast-targeted DnaJ-like protein localized to the plastid envelope membrane. An ARC6-green fluorescent protein fusion protein was localized to a ring at the center of the chloroplasts and rescued the chloroplast division defect in the arc6 mutant. The ARC6 gene product is related closely to Ftn2, a prokaryotic cell division protein unique to cyanobacteria. Based on the FtsZ filament morphology observed in the arc6 mutant and in plants that overexpress ARC6, we hypothesize that ARC6 functions in the assembly and/or stabilization of the plastid-dividing FtsZ ring. We also analyzed FtsZ localization patterns in transgenic plants in which plastid division was blocked by altered expression of the division site-determining factor AtMinD. Our results indicate that MinD and ARC6 act in opposite directions: ARC6 promotes and MinD inhibits FtsZ filament formation in the chloroplast. PMID- 12897265 TI - Somatization disorders in children and adolescents. PMID- 12897263 TI - Two-step regulation and continuous retrotransposition of the rice LINE-type retrotransposon Karma. AB - Here, we report the identification of Karma, a LINE-type retrotransposon of plants for which continuous retrotransposition was observed in consecutive generations. The transcription of Karma is activated in cultured cells of rice upon DNA hypomethylation. However, transcription is insufficient for retrotransposition, because no increase in the copy number was observed in cultured cells or in the first generation of plants regenerated from them. Despite that finding, copy number increase was detected in the next generation of regenerated plants as well as in later generations, suggesting that the post transcriptional regulation of Karma retrotransposition is development dependent. Our results indicate that two different mechanisms, one transcriptional and the other developmental, control the mobilization of KARMA: In addition, unlike other known active plant retrotransposons, Karma is not subject to de novo methylation, and retrotransposition persists through several generations. PMID- 12897266 TI - Evidence-based medicine: asking the answerable question (question templates as tools). PMID- 12897267 TI - Consultation with the specialist: palpitations, syncope, and sudden cardiac death in children: who's at risk? PMID- 12897268 TI - Index of suspicion. PMID- 12897269 TI - Question from the clinician: racemic epinephrine for bronchiolitis. PMID- 12897270 TI - Developing a statewide network of family resource centers in New Hampshire: lessons learned. PMID- 12897271 TI - Bonding. PMID- 12897272 TI - Otitis media and tympanostomy tube insertion during the first three years of life: developmental outcomes at the age of four years. AB - OBJECTIVE: In a long-term, prospective study, we set out to determine whether otitis media in the first 3 years of life persisting for periods currently considered developmentally threatening actually results in later impairments of children's cognitive, language, speech, or psychosocial development; whether prompt insertion of tympanostomy tubes prevents or lessens any such impairments; and whether, irrespective of causality, associations exist between persistent early-life otitis media and later developmental impairments. This report describes findings in study participants at the age of 4 years. METHODS: We enrolled 6350 healthy infants from 2 to 61 days of age at urban hospitals and 2 small-town/rural and 4 suburban private pediatric practices. We regularly evaluated the children for the presence of middle-ear effusion (MEE) throughout their first 3 years of life by pneumatic otoscopy, supplemented by tympanometry; we monitored the validity of the otoscopic observations on an ongoing basis; and we treated children for otitis media according to specified guidelines. In the clinical trial component of the study, we randomly assigned 429 children who met specified minimum criteria regarding the persistence of MEE to undergo tympanostomy tube insertion either promptly or after a defined extended period if MEE remained present. In the associational component of the study, we selected a representative sample of 241 children who ranged from having no MEE to having MEE the cumulative duration of which fell just short of meeting randomization criteria for the clinical trial. In 397 (92.5%) of the children in the clinical trial and in 234 (97.1%) of the children in the representative sample, we assessed cognitive, language, speech, and psychosocial development at the age of 4 years, using formal tests, conversational samples, and parent questionnaires. RESULTS: In children in the randomized clinical trial, there were no statistically significant differences in mean (+/-standard deviation) scores (higher denotes more favorable) favoring the early-treatment group over the late treatment group on the General Cognitive Index of the McCarthy Scales of Children's Abilities (97 +/- 14 and 98 +/- 14, respectively); the Peabody Picture Vocabulary Test-Revised, a measure of receptive language (90 +/- 15 vs 92 +/- 16); the Nonword Repetition Test, a measure of phonological memory (66 +/- 12 vs 70 +/- 12); the Number of Different Words, a measure of word diversity (150 +/- 34 vs 150 +/- 31); the Mean Length of Utterance in Morphemes, a measure of sentence length and grammatical complexity (3.4 +/- 0.8 vs 3.4 +/- 0.7); or the Percentage of Consonants Correct-Revised, a measure of speech-sound production (92 +/- 5 vs 93 +/- 5). There were also no significant differences in ratings (higher denotes less favorable) on the Parenting Stress Index-Short Form (Total Stress scores: 68 +/- 18 vs 65 +/- 17) or the Child Behavior Checklist (Total Problem T scores: 50 +/- 10 vs 49 +/- 10). In the associational component of the study, correlations between the children's durations of MEE and their developmental outcomes were generally weak and, in most instances, nonsignificant. Exceptions, after adjustment for sociodemographic variables and for hearing thresholds at the time of developmental testing, consisted of a significant negative correlation between children's cumulative durations of MEE in their first 3 years of life and scores on the McCarthy Verbal subscale, and significant positive correlations between durations of MEE and scores on 2 measures of parent-child stress. The percentage of variance in these scores explained by time with MEE beyond that explained by sociodemographic variables ranged from 1.6% to 3.3%. In both the randomized clinical trial and the associational component, sociodemographic variables seemed to be the most important factors influencing developmental outcomes, and in both components, the results at 4 years of age were consistent with the results that had been obtained at 3 years of age. CONCLUSIONS: In otherwise healthy children who are younger than 3 years and have persistent MEE within the duration limits that we studied, prompt insertion of tympanostomy tubes does not measurably improve developmental outcomes at 4 years of age. In such children, persistent MEE within the duration limits that we studied is negligibly associated with and probably does not affect developmental outcomes at 4 years of age. PMID- 12897273 TI - Length of stay for common pediatric conditions: teaching versus nonteaching hospitals. AB - OBJECTIVE: Pediatric teaching hospitals provide particular expertise in caring for children with complex or severe illnesses, yet most patients within teaching hospitals have common pediatric conditions. No study has determined whether children with common conditions remain hospitalized at teaching institutions longer than at nonteaching institutions. The objective of this study was to compare length of stay (LOS) for common pediatric conditions between teaching and nonteaching hospitals. METHODS: This study uses Massachusetts's hospital data for all discharged children ages 0 to 17 years for 1995 and 1996. Discharges were included when the principal diagnosis indicated asthma, bacterial pneumonia, convulsions, dehydration, failure to thrive, gastroenteritis, or urinary tract infections. Hospitals were classified as either teaching or nonteaching using the 1995-1996 American Hospital Association Guide. Children were identified as having a chronic condition when any discharge diagnosis was 1 of those on a previously published catalog of chronic childhood illnesses. The analysis tested the association of hospital type with LOS, controlling for chronic conditions, insurance type, age, race, diagnosis, mortality, and disposition using multivariate linear regression. RESULTS: Of 17 890 discharges for a common pediatric condition during the study period, 52.3% were from teaching hospitals. Twelve percent of common condition discharges also had a chronic disease diagnosis; 75.1% of these were discharged from a teaching hospital. LOS from nonteaching hospitals was shorter than from teaching hospitals (2.42 days vs 3.20 days). Although LOS for stays with a chronic diagnosis were longer than those without (4.75 days vs 2.56 days), controlling for chronic illness and other covariates did not eliminate the difference between LOS for nonteaching hospitals versus teaching hospitals (1.65 days vs 2.23 days). CONCLUSION: Pediatric patients with common conditions have a shorter LOS in nonteaching hospitals than those admitted to teaching hospitals by a little more than half a day. These results are unchanged when accounting for chronic conditions despite the expected results of preferential admissions to teaching hospitals for this group of patients. Additional studies should better characterize differences in patient populations, describe differences in processes, and identify differences in patient experience and outcomes to understand better the potential benefits of treating children with specific conditions at particular types of hospitals. PMID- 12897274 TI - Prevalence of serious bacterial infections in febrile infants with respiratory syncytial virus infection. AB - OBJECTIVE: Neonates with fever generally undergo a full, invasive septic evaluation to exclude serious bacterial infection (SBI). The risk of SBI in febrile older infants and children with documented respiratory syncytial virus (RSV) infection has been found to be negligible. The purpose of this study was to investigate the prevalence of SBI in febrile infants who were younger than 8 weeks and had documented RSV infection and to compare the risk of SBI with control subjects who were febrile and RSV-negative. METHODS: This was a retrospective cohort study of infants who were age 8 weeks or less and presented with documented fever to the emergency department at an urban children's hospital in October through April during a 4-year period. RSV-positive cases were gender- and age-matched to febrile RSV-negative control subjects. Clinical characteristics and the rate of SBI were compared between the 2 groups. RESULTS: A total of 174 previously healthy infants with fever and a positive RSV antigen test were identified and matched with 174 previously healthy infants with fever and a negative RSV test. Infants with RSV infection were more likely to present with upper respiratory infection symptoms, increased work of breathing, and apnea. Overall, 2 patients in the RSV group had SBI (both with urinary tract infections), compared with 22 in the control group (relative risk: 0.009), 17 of which were urinary tract infections. CONCLUSIONS: The risk of SBI in febrile infants with RSV infection seems to be very low, particularly in comparison with a control group of RSV-negative infants. These data suggest that full septic evaluations are not necessary in nontoxic-appearing infants with a positive RSV test. It seems to be prudent to examine the urine in these infants, as there is a clinically relevant rate of urinary tract infection. PMID- 12897275 TI - Sequelae of severe respiratory syncytial virus infection in infancy and early childhood among Alaska Native children. AB - OBJECTIVE: In 1993-1996, we conducted a nested case-control study to determine risk factors for hospitalization with respiratory syncytial virus (RSV) infection among Alaska Native infants and young children. In the current study, we returned to former RSV case-patients and their control subjects during 1999-2001 to determine whether children who are hospitalized with RSV at <2 years of age are more likely to develop chronic respiratory conditions. METHODS: For each former RSV case-patient and control subject from remote villages in southwest Alaska, we reviewed medical records, interviewed parents, performed physical examinations and spirometry, collected sera, and analyzed chest radiographs. Case-patients were identified through surveillance for RSV hospitalization, and matched control subjects without lower respiratory infection (LRI)-related hospitalization were identified. RESULTS: Hospitalization for RSV infection was associated with a significant increase in wheezing, LRIs, and asthma diagnosis during the first 4 years of life. The association decreased with age and was no longer significant by 5 years of age. However, hospitalization for RSV infection was associated with increased respiratory symptoms and increased chronic productive cough at 5 to 8 years of age. Children who were hospitalized with RSV were not more likely at follow-up to have allergies, eczema, or a positive family history of asthma. CONCLUSIONS: Severe RSV infection in infancy may produce airway injury, which is manifested in chronic productive cough with or without wheezing and recurrent LRIs. Although the association of RSV infection with wheezing seems to be transient, children remain at higher risk for chronic productive cough at 5 to 8 years of age. RSV prevention modalities may prevent sequelae that occur early and later in childhood. PMID- 12897276 TI - Office preparedness for pediatric emergencies: a randomized, controlled trial of an office-based training program. AB - OBJECTIVE: Many children enter the emergency medical system through primary care offices, yet these offices may not be adequately prepared to stabilize severely ill children. We conducted this study to evaluate the effectiveness of an office based educational program designed to improve the preparation of primary care practices for pediatric emergencies. METHODS: A prospective, randomized, controlled trial was conducted of primary care practices (pediatric, family practice, and health departments) that were recruited from an existing database of North Carolina practices. Practices that agreed to participate were randomly assigned to either the intervention or the control group. Unannounced mock codes were conducted in the intervention practices by 2 emergency medicine clinicians (medical doctor and/or registered nurse). Practices were expected to respond to the mock code using their own staff, equipment, and local emergency medical system. After the exercise, the emergency medicine clinicians and the local emergency medical system team led a structured debriefing session providing constructive feedback to the staff on their performance, a review of the office's equipment, and a resource manual designed for the project. The primary outcome measures were obtained by survey 3 to 6 months postintervention and included 1) purchase of new pediatric emergency equipment and medications, 2) receipt or updating of basic life support/pediatric advanced life support/advanced life support training by staff members, and 3) development of written emergency pediatric protocols. The control practices received no interventions during the trial and completed a similar outcome survey. RESULTS: Thirty-nine practices (20 intervention, 19 control) completed the trial. There were no significant differences in practice characteristics between the 2 groups. Intervention practices were more likely to develop written office protocols (60% vs 21%); more staff in the intervention practices received additional basic life support/pediatric advanced life support/advanced life support training 3 to 6 months after the intervention (118 vs 54). There were no significant differences in the purchase of new equipment or medications. Ninety percent of the intervention practices rated the intervention as useful for their practice, and 95% believed that the program should be continued. CONCLUSIONS: The findings suggest that the intervention was well received and motivated practices to take concrete actions to prepare for pediatric emergencies. PMID- 12897277 TI - Resuscitation of newborn infants with 21% or 100% oxygen: follow-up at 18 to 24 months. AB - OBJECTIVE: To follow-up children who had been resuscitated at birth with either 21% or 100% oxygen (O2). METHODS: A multicenter study with 10 participating centers recruited 609 infants to the Resair 2 study where resuscitation was performed with either 21% or 100% O2. A follow-up between ages 18 and 24 months was performed. However, during follow-up registration, it was found that 18 infants had been enrolled twice in the original Resair 2 study with different registration numbers, leaving 591 enrolled in the Resair 2 study and 410 enrolled in the 7 centers participating in the follow-up. Of these 410 infants, 79 died (76 in the neonatal and 3 in the postneonatal period). Furthermore, for 8 infants informed consent was not obtained, leaving 323 eligible for follow-up. Of these, 213 infants (66%) were followed-up: 91 (62%) had been resuscitated with 21% O2, and 122 (69%) with 100% O2. At a median age of 22 and 20 months (not significant) in the 21% and 100% groups, respectively, a simple questionnaire was filled out and neurologic assessment was performed in addition to measuring anthropometric data. RESULTS: There were no significant differences in weight, height, or head circumference between the 2 groups. Cerebral palsy developed in 10% and 7%, respectively, in the 2 groups (not significant). In total, 11 cases (12%) in the 21% versus 11 cases (9%) in the 100% O(2) group (odds ratio: 1.39, 95% confidence interval: 0.57-3.36) developed cerebral palsy and/or mental or other delay. Furthermore, it was concluded that 14 (15%) in the 21% group and 12 (10%) in the 100% group were not normal (odds ratio: 1.67, 95% confidence interval: 0.73 3.80). CONCLUSIONS: There were no significant differences in somatic growth or neurologic handicap at an age of 18 to 24 months in infants resuscitated with either 21% or 100% O2 at birth. Based on these data, resuscitation with ambient air seems to be safe, at least in most cases. More studies are needed to settle this issue. PMID- 12897278 TI - Learning, cognitive, and attentional problems in adolescents born small for gestational age. AB - OBJECTIVE: To determine whether the presence, severity, or symmetry of growth restriction in term infants is an independent risk factor for learning, cognitive, and attentional problems in adolescence. METHODS: A total of 7388 term infants have been followed prospectively since birth. At 14 years, 5059 mothers completed a Child Behavior Checklist and provided information on their child's school progress. A total of 5051 adolescents completed a Youth Self Report, with 3703 also undergoing psychometric testing with Ravens Progressive Matrices and Wide Range Achievement Test (WRAT) reading subtest. Outcomes were compared on the basis of birth weight groups and measures of body symmetry and were adjusted for the level of social risk at birth. RESULTS: Adolescents who were born small for gestational age (SGA), when compared with their appropriately grown counterparts (>10th percentile), were more likely to experience learning difficulties, with a higher prevalence in those of birth weight < or =3rd percentile. Girls of birth weight < or =3rd percentile were more likely to have attentional problems and low WRAT reading scores. There was no significant difference in Ravens IQ or mean WRAT reading scores between SGA and non-SGA groups. There was no association between body symmetry and any of the outcomes studied. CONCLUSIONS: SGA status seems to have only modest independent effects on learning, cognition, and attention in adolescence. Severity but not symmetry of growth restriction predicted learning difficulties. PMID- 12897279 TI - Parental evaluation of informing interviews for cleft lip and/or palate. AB - OBJECTIVE: This study was conducted to record subjective information concerning parental experiences and preferences with regard to informing interviews for cleft lip and/or palate. The analysis of these data was intended to help informants improve the way in which such conversations are conducted. METHODS: The study used a self-administered questionnaire. Study participants were the biological parents of children who had cleft lip and/or palate and were seen in the University of Pittsburgh Cleft Palate-Craniofacial Center. RESULTS: Results revealed that parents wanted informing health professionals to be in greater control of the informing conversation, to show more caring and confidence, to show more of their own feelings, to give parents more of an opportunity to talk and show feelings, to make a greater effort to comfort parents, to provide more information, to initiate more of a discussion about the association between clefts and mental retardation/learning disabilities, and to provide more referrals to other parents during the informing interview. A positive association between the degree to which parents reported knowing the health professionals and reported satisfaction was identified for 10 dimensions of health professional behavior. CONCLUSIONS: Parents are dissatisfied with several aspects of informing interviews for cleft lip and/or palate. The results of this study suggest ways to improve informing interviews for oral-facial clefts and other congenital anomalies. PMID- 12897280 TI - Children's health insurance status and emergency department utilization in the United States. AB - OBJECTIVES: Emergency department (ED) overcrowding has become a national problem. Children account for nearly 25% of overall ED visits. It has been reported that uninsured and publicly insured children are likely to visit the ED for urgent and nonurgent problems, yet it remains unclear to what extent health insurance status would influence children's overall ED utilization or ED utilization for nonurgent problems at the national level after controlling for other confounding factors. The objective of this study was to examine the effect of health insurance status on children's overall ED utilization and children's ED utilization for nonurgent problems among the general pediatric population in the United States. METHODS: Secondary analysis of the household component of the 1997 Medical Expenditure Panel Survey on 10 193 children younger than 18 years. The main outcome measures were annual overall ED utilization and ED utilization for nonurgent problems. RESULTS: During 1997, 10.8% of children were uninsured for the entire year. A total of 17.5% of children were publicly insured the entire year, whereas 55.3% of children held private insurance the entire year. There were also 16.5% of children who were insured only part of the year. Without adjusting for covariates, publicly insured children were more likely to have an ED visit during the year than both privately insured children (unadjusted odds ratio [OR]: 1.26; 95% confidence interval [CI]: 1.03-1.55) and uninsured children (unadjusted OR: 1.46; 95% CI: 1.1-1.95). The difference between publicly insured and privately insured children (adjusted OR: 0.90; 95% CI: 0.70-1.16) and between publicly insured and uninsured children (adjusted OR: 1.12; 95% CI: 0.84-1.49) became insignificant after controlling for covariates. With or without adjustments for covariates, there was no significant difference in the likelihood of having an ED visit between privately insured and uninsured children. Similar to the utilization pattern of overall ED visits, publicly insured children were more likely to have a nonurgent ED visit than both privately insured (unadjusted OR: 1.86; 95% CI: 1.36-2.53) and uninsured children (unadjusted OR: 1.81; 95% CI: 1.15-2.84). Both differences disappeared after controlling for covariates. There was no significant difference in the likelihood of nonurgent ED visits between privately insured and uninsured children with or without adjustments for covariates. CONCLUSIONS: Health insurance status was not associated with children's overall ED use or children's ED use for nonurgent problems at the national level. Our findings suggest that policy efforts in an attempt to relieve ED overcrowding conditions should look for measures beyond solely making changes in health insurance coverage for children. PMID- 12897281 TI - Bicycle helmet assessment during well visits reveals severe shortcomings in condition and fit. AB - BACKGROUND: Improper bicycle helmet fit increases the risk of head injury. Information on the rate of proper use of bicycle helmets is lacking. Promotion of helmet use is recommended at well-child and adolescent visits. Actual helmet assessment during such visits has not been reported. OBJECTIVES: The primary goal of this study is to measure the proportion of children whose helmets are in proper condition and can be made to fit properly by the child and/or parent. The secondary goal is to begin to assess the value and practicality of helmet inspection during well-child and adolescent visits. METHODS: The study took place at a private pediatric office in Falmouth, Massachusetts, from June 1 through August 31, 2001. Eligible children and adolescents were those aged 4 to 18 years presenting for well examination, along with siblings present at the visit. Eligible families completed a questionnaire, then had a timed attempt to fit a helmet, followed by an assessment of helmet fit and condition against a predetermined standard. RESULTS: Eighty-four percent (395/473) of eligible families participated. A total of 479 participants were assessed. Eighty-eight percent of participants (419/478) owned a helmet. Reported helmet use "always" or "almost always" was 73% for bicycling (317/434), 69% for in-line skating (193/279), 58% for scootering (179/310), and 50% for skateboarding (79/158). Compared with younger children, teenagers were less likely to wear helmets for all activities. Complete pass rate for every aspect of condition and fit was 4% (20/478, 95% confidence interval: 3-6). The pass rate when the parent alone fit the helmet was 0% (0/52). Three individual aspects of fit were most problematic: 1) helmet 'resting position' too high on the forehead (pass rate 249/479; 52%), 2) improper strap position (pass rate 157/476; 33%), and 3) excessive movement of the helmet from front to back of the head (pass rate 247/479; 52%). Mean time for questionnaire completion was 4 (standard deviation: +/-1) minutes, and 7 (standard deviation: +/-3) minutes for helmet assessment. CONCLUSIONS: Ninety-six percent of children and adolescents wore helmets in inadequate condition and/or with inadequate fit. This occurred despite a high acceptance of helmet use by this population. Initial evidence suggests that helmet assessment during well visits may be practical and valuable. PMID- 12897282 TI - Transcranial Doppler ultrasonography and neurocognitive functioning in children with sickle cell disease. AB - OBJECTIVE: We examined the relationship between cerebral blood flow velocity, measured by transcranial Doppler (TCD) ultrasonography, and neurocognitive functioning. METHODS: Participants were 60 children who had sickle cell disease (HbSS) and had no documented history of stroke. Children were classified according to Stroke Prevention Trial in Sickle Cell Anemia criteria (normal, conditional, and abnormal), and their performance was compared on measures of intellectual abilities, academic achievement, sustained attention/concentration, executive function, and parent and teacher ratings of executive function. RESULTS: Children with abnormal TCD values performed more poorly than children with conditional TCD values on measures of verbal intelligence and executive function. Children with conditional TCD values performed more poorly than children with normal TCD values on measures of sustained attention/concentration and executive function. TCD values also were a significant predictor of auditory working memory in exploratory analyses. CONCLUSIONS: Our findings support the hypothesis that neurocognitive functions subserved by the frontal systems (eg, sustained attention/concentration and executive function) seem to be the most useful indices of progressive cerebrovasculopathy in children with HbSS disease. PMID- 12897283 TI - The natural course of infantile Pompe's disease: 20 original cases compared with 133 cases from the literature. AB - OBJECTIVE: Infantile Pompe's disease is a lethal cardiac and muscular disorder. Current developments toward enzyme replacement therapy are promising. The aim of our study is to delineate the natural course of the disease to verify endpoints of clinical studies. METHODS: A total of 20 infantile patients diagnosed by the collaborative Dutch centers and 133 cases reported in literature were included in the study. Information on clinical history, physical examination, and diagnostic parameters was collected. RESULTS: The course of Pompe's disease is essentially the same in the Dutch and the general patient population. Symptoms start at a median age of 1.6 months in both groups. The median age of death is 7.7 and 6 months, respectively. Five percent of the Dutch patients and 8% of all reported patients survive beyond 1 year of age. Only 2 patients from literature became older than 18 months. A progressive cardiac hypertrophy is characteristic for infantile Pompe's disease. The diastolic thickness of the left ventricular posterior wall and cardiac weight at autopsy increase significantly with age. Motor development is severely delayed and major developmental milestones are generally not achieved. For the Dutch patient group, growth deviates significantly from normal despite start of nasogastric tube feeding. Levels of aspartate aminotransferase, alanine aminotransferase, lactate dehydrogenase, creatine kinase, or creatine kinase-myocardial band isoenzyme are typically elevated, although aspartate aminotransferase, alanine aminotransferase, and lactate dehydrogenase increase significantly with age. The patients have fully deleterious mutations. Acid alpha-glucosidase activity is severely deficient. CONCLUSIONS: Survival, decrease of the diastolic thickness of the left ventricular posterior wall, and achievement of major motor milestones are valid endpoints for therapeutic studies of infantile Pompe's disease. Mutation analysis and measurement of the alpha-glucosidase activity should be part of the enrollment program. PMID- 12897284 TI - Cadaver-donor renal transplantation of children in Israel (1990-2001): racial disparities in health care delivery? AB - OBJECTIVE: To evaluate the allocation and outcome of cadaver-donor renal transplantation (CDRTx) among Jewish and Arab children in Israel. METHODS: Data on CDRTxs in patients who had end-stage renal failure (ESRF), were younger than 18 years, and were on dialysis treatment were obtained for the 11-year period of January 1990 to December 2000 from the Israeli Dialysis and Transplant Registry, supplemented by 10 years of follow-up (January 1991-December 2000) from our own center. RESULTS: The Israeli Dialysis and Transplant Registry data show that 64 of 130 available cadaver-donor kidneys (CDKs) were allocated to Jewish patients (49.2%) and 66 of 130 were allocated to Arab children (50.8%): Moslem, Druze, or Christian. The Jew/Arab patient ratio for a waiting time of <1 year was 0.97 and for 1 to 2 years was 1.45, whereas that ratio was 0.6 for 2 to 4 years and 2.0 for >4 years. The mean renal transplant score (RTx score), reflecting the urgency of the need for RTx of an ESRF patient, was identical for Jew and Arab: 4.93 and 4.96. Our own center data refer to 69 dialysis (47 Arabs and 22 Jews) and 4 predialysis patients younger than 18 years who underwent 78 RTxs. Eighteen Arab and 14 Jewish children from our center received 20 and 15 CDRTxs in Israel, with a mean waiting time of 29.6 and 25.4 months for Jew and Arab, respectively (ratio: 1.16). In our center, the outcome (after 7 years) of graft survival and function was not different between Jewish and Arab RTx recipients. CONCLUSIONS: Allocation of CDRTxs between young Jewish and Arab ESRF patients on dialysis did not differ and was associated with comparable waiting times, identical RTx scores, and similar long-term outcome. This is a remarkable finding, certainly in the face of the unequal race allocation of RTxs in the United States as well as the long unstable local (Middle East) political situation. PMID- 12897285 TI - Randomized trial of prolonged low-dose versus conventional-dose indomethacin for treating patent ductus arteriosus in very low birth weight infants. AB - OBJECTIVE: Indomethacin is used for closing the patent ductus arteriosus in premature infants. Prolonged low-dose indomethacin given over 6 days could potentially improve closure rates because ductal constriction is maintained long enough for more effective anatomic closure. We compared the efficacy of this regimen to conventional dosing in a cohort of very low birth weight infants. METHODS: In a 2-arm clinical trial, 140 infants were randomized to either conventional dose (0.2 mg/kg/dose every 12 hours for 3 doses) or prolonged low dose indomethacin (0.1 mg/kg/dose daily for 6 doses). The primary outcome measure was ductal closure rate, and the secondary outcomes were the need for a second course of treatment, surgical ligation rates, and side effects. RESULTS: Ductal closure after 1 course of indomethacin was similar between the 2 groups: 68% for the conventional dose group and 72% for the prolonged low dose (mean difference 4%; 95% confidence interval: -19% to 11%). The incidence of transient oliguria was higher in the conventional dose group, 31% versus 9%. There was a trend toward more necrotizing enterocolitis in the prolonged low-dose group, 7.0% versus 1.4%. CONCLUSIONS: There was no difference in efficacy between the 2 dosing regimens. In view of this and with its higher incidence of necrotizing enterocolitis, we do not recommend using prolonged low-dose indomethacin for closing the patent ductus arteriosus in very low birth weight infants. PMID- 12897286 TI - Mortality and neurologic, mental, and psychomotor development at 2 years in infants born less than 27 weeks' gestation: the Leiden follow-up project on prematurity. AB - OBJECTIVE: To determine the outcome of infants with a gestational age (GA) <27 weeks, born in the mid-1990s. DESIGN: Regional, prospective study; part of the Leiden Follow-Up Project on Prematurity. SETTING: Three health regions in The Netherlands. PATIENTS: A total of 266 live born infants (1996/1997) with GA <32 weeks; 46 infants were <27 weeks. MAIN OUTCOME MEASURES: Neurologic examination (according to Hempel) and assessment of mental and psychomotor development using the Bayley Scales of Infant Development I, at the corrected age of 2 years. RESULTS: Mortality was 35% (16 of 46) <27 weeks, compared with 6% (14 of 220) in infants with GA 27 to 32 weeks; withdrawal of treatment in 60% and 43%, respectively. Below 27 weeks mortality was higher after extra-uterine transport and pregnancy induction. Neonatal morbidity was higher in infants <27 weeks compared with infants 27 to 32 weeks. Below 27 weeks postnatal use of dexamethasone and being hospitalized at term were associated with abnormal neurologic outcome; there was a higher incidence in (mild) mental developmental delay compared with the older infants. Adverse outcome (dead or abnormal neurologic, psychomotor, or mental development) in infants 23 to 24, 25, 26, and 27 to 32 weeks GA was, respectively, 92% (11 of 12), 64% (7 of 11), 35% (8 of 23), and 18% (40 of 220). CONCLUSIONS: Mortality and neonatal morbidity were higher in infants with GA <27 weeks compared with infants born between 27 and 32 weeks. The high adverse outcome of infants <25 weeks suggests that one should carefully weigh whether or not to aggressively resuscitate and treat these extremely premature infants. PMID- 12897287 TI - Clinical tolerance to lactose in children with cow's milk allergy. AB - OBJECTIVE: Adverse reactions following the ingestion of lactose have been reported in children with cow's milk (CM) allergy. Whether this is attributable to the contamination of lactose with CM proteins is unknown. In this paper, we assessed clinical tolerance of lactose derived from CM whey in children hypersensitive to CM from 2 university hospital pediatric departments. DESIGN: Twenty-four children (5 girls and 19 boys, median 25 months old; range: 2-107 months) with immediate CM allergy confirmed at history or during double-blind, placebo-controlled food challenge (DBPCFC) were enrolled. DBPCFC with CM could be conducted in 11 of 24 patients. Children with a history of immediate/delayed reactions to soy formula (SF) were excluded. Clinical tolerance to CM, SF, and SF + lactose was assessed by: 1) skin prick test with casein, lactalbumin, soy commercial allergen preparations, fresh CM, SF, SF and lactose, lactose (Official Pharmacopoeia) in 4 concentrations (0.01%, 0.1%, 1%, 10%); 2) specific serum immunoglobulin E determination by CAP system technology; 3) DBPCFC in 8 incremental doses of SF + lactose and using SF as a placebo to make up a total of 240 mL of reconstituted formula. RESULTS: With a positive cutoff point of > or = 3 mm wheal diameter at SPT, all patients were sensitized to fresh CM, lactalbumin, and/or casein. Twenty-three of 24 patients (95.8%) were SPT-positive to CM formula, 16 of 24 to lactalbumin (66.6%), 14 of 24 to casein (58.3%), and none to SF, SF + lactose, or lactose alone at all dilutions. Complexed immunoglobulin E determinations were positive for CM in 23 of 24 cases and negative in all cases for soy. Challenge with SF + lactose was negative in all cases. CONCLUSIONS: Even children hypersensitive to CM are clinically tolerant to lactose and can safely consume foods and drugs with lactose from bovine sources as an ingredient. Lactose exclusion is unwarranted from soy preparations on grounds of potential allergic reactions to CM protein residue. PMID- 12897288 TI - Impact of the rapid diagnosis of influenza on physician decision-making and patient management in the pediatric emergency department: results of a randomized, prospective, controlled trial. AB - OBJECTIVE: To determine the impact of the rapid diagnosis of influenza on physician decision-making and patient management, including laboratory tests and radiographs ordered, patient charges associated with these tests, antibiotics/antivirals prescribed, and length of time to patient discharge from the emergency department. METHODS: Patients aged 2 months to 21 years presenting to an urban children's teaching hospital emergency department were screened for fever and cough, coryza, myalgias, headache, and/or malaise. After obtaining informed consent, patients were randomized to 1 of 2 groups: 1) physician receives (physician aware of) the rapid influenza test result; or 2) physician does not receive (physician unaware of) the result. For patients in the physician aware group, nasopharyngeal swabs were obtained, immediately tested with the FluOIA test for influenza A and B, and the result was placed on the chart before patient evaluation by the attending physician. For the physician unaware group, nasopharyngeal swabs were obtained, stored according to manufacturer's directions, and tested within 24 hours. Results for the physician unaware group were not disclosed to the treating physicians at any time. The 2 resultant influenza-positive groups (aware and unaware) were compared for laboratory and radiograph studies and their associated patient charges, antibiotic/antiviral prescriptions, and length of stay in the emergency department. RESULTS: A total of 418 patients were enrolled, and 391 completed the study. Of these, 202 tested positive for influenza. Comparison of the 96 influenza-positive patients whose physician was aware of the result with the 106 influenza-positive patients whose physician was unaware of the result revealed significant reductions among the former group in: 1) numbers of complete blood counts, blood cultures, urinalyses, urine cultures, and chest radiographs performed; 2) charges associated with these tests; 3) antibiotics prescribed; and 4) length of stay in the emergency department. The number of influenza-positive patients who received prescriptions for antiviral drugs was significantly higher among those whose physician was aware of the result. CONCLUSIONS: Physician awareness of a rapid diagnosis of influenza in the pediatric emergency department significantly reduced the number of laboratory tests and radiographs ordered and their associated charges, decreased antibiotic use, increased antiviral use, and decreased length of time to discharge. PMID- 12897289 TI - Medicaid participation by private and safety net pediatricians, 1993 and 2000. AB - BACKGROUND: Eligibility expansions and managed care growth were 2 major forces shaping the Medicaid program during the 1990s. Although Medicaid managed care was introduced to contain rising costs of growing enrollment and expenditures, it also offered states an opportunity to improve enrollees' access to mainstream health care providers. By enrolling in commercial managed care plans, they could gain access to private office-based physicians, thus eliminating a 2-tiered health care system. OBJECTIVES: To investigate changes in private and safety net pediatricians' participation in Medicaid between 1993 and 2000, a period noted for eligibility expansion for children and rapid managed care growth. DESIGN: Survey data collected from private and safety net pediatricians in 1993 and 2000 were analyzed to investigate how pediatricians' Medicaid caseloads were affected by 1) their practice setting, and 2) whether they accepted all Medicaid patients. RESULTS: Pediatricians' Medicaid caseloads increased significantly between 1993 and 2000, with those in private practice settings reporting greater increases. More pediatricians accepted all Medicaid patients in 2000, but Medicaid caseloads remained twice as high for safety net pediatricians. Medicaid caseloads increased dramatically for those accepting all Medicaid patients, except in safety net settings, where pediatricians who were turning away some Medicaid patients had heavier Medicaid caseloads than those who accepted all Medicaid patients. CONCLUSIONS: Although Medicaid children were still disproportionately served in the safety net in 2000, their access to private pediatricians had improved since 1993. But a ceiling in the safety net's capacity to absorb more Medicaid patients was suggested by the finding that safety net pediatricians turned away Medicaid patients as their Medicaid caseloads increased. Combined with decreasing participation by commercial plans in the Medicaid managed care market, increasing hospital emergency department overloads, and growing Title XXI Medicaid enrollments, interventions may be needed to avert a looming pediatric provider shortage for publicly insured low-income children. PMID- 12897290 TI - Developing circadian rhythmicity in infants. AB - Circadian rhythms are endogenously generated rhythms with a period length of approximately 24 hours. Evidence gathered during the past decade indicates that the circadian timing system develops prenatally and the suprachiasmatic nuclei, the site of a circadian clock, is present by midgestation in primates. Recent evidence also shows that the circadian system of primate infants is responsive to light at very premature stages and that low-intensity lighting can regulate the developing clock. After birth, there is progressive maturation of the circadian system outputs, with pronounced rhythms in sleep-wake and hormone secretion generally developing after 2 months of age. Showing the importance of photic regulation of circadian phase in infants, exposure of premature infants to low intensity cycled lighting results in the early establishment of rest-activity patterns that are in phase with the 24-hour light-dark cycle. With the continued elucidation of circadian system development and influences on human physiology and illness, it is anticipated that consideration of circadian biology will become an increasingly important component of neonatal care. PMID- 12897291 TI - Treating exacerbations of asthma in children: the role of systemic corticosteroids. AB - OBJECTIVE: To review the use of systemic corticosteroids to treat recurrent, acute asthma episodes in children, with a focus on the role of oral corticosteroids. METHODS: A comprehensive review of the literature was performed using the Medline database (January 1966-October 2002) and the Embase database (January 1980-August 2002). RESULTS: The significant findings of 17 selected, controlled clinical trials of oral corticosteroids (OCSs) for acute exacerbations of asthma in children, compared with placebo or with other formulations of corticosteroids, can be summarized as follows: 1) OCSs are effective for the outpatient treatment of acute asthma, 2) pulmonary function tests may not be the best means of assessing the efficacy of OCSs for acute asthma, 3) early administration of OCSs for acute asthma reduces hospitalizations, 4) the critical factor for a positive outcome is early administration of the corticosteroid, and 5) OCSs are preferred for the outpatient treatment of acute asthma. CONCLUSIONS: Early treatment of acute asthma symptoms with OCSs in children with a pattern of recurrent acute asthma may decrease the severity of acute asthma episodes and reduce the likelihood of subsequent relapses. Attention should be given to identifying these children and standardizing a treatment approach based on accepted, consistent definitions of what constitutes an asthma exacerbation and recurrence. A suggested protocol is described. PMID- 12897292 TI - The science and fiction of the "resurgence" of pertussis. PMID- 12897293 TI - Pediatric advocacy: yesterday, today, and tomorrow. PMID- 12897294 TI - Racial disparities in renal transplantation in children. PMID- 12897295 TI - Resident stress revisited: a senior pediatric resident's point of view. PMID- 12897297 TI - Replicating measurements. PMID- 12897298 TI - The effect of Medicaid participation by private and safety net pediatricians on incremental expansion of coverage for children. PMID- 12897299 TI - Toward a quality workforce. PMID- 12897301 TI - The pediatric generalist and integrative care. PMID- 12897302 TI - Recombinant granulocyte colony-stimulating factor administered enterally to neonates is not absorbed. AB - Granulocyte colony-stimulating factor (G-CSF) is present in liquids swallowed by the fetus and neonate; specifically, amniotic fluid, colostrum, and human milk. The swallowed G-CSF has local effects on enteric cells, which express the G-CSF receptor. However, some portion of the G-CSF ingested by the fetus and neonate might be absorbed into the circulation and have systemic actions, such as stimulating neutrophil production. To assess this possibility we sought to determine if circulating G-CSF concentrations of neonates increase after enteral administration of recombinant human granulocyte colony-stimulating factor (rhG CSF). This was a single-center, prospective, blinded, randomized, 2 x 2 crossover study, with each infant receiving 1 dose of rhG-CSF (100 microg/kg) and 1 dose of placebo. Plasma G-CSF concentrations were measured at 2 and 4 hours after administration of the test solution. No significant change in plasma G-CSF concentration was observed after the enteral administration of rhG-CSF. On this basis, we conclude that orally administered rhG-CSF is not absorbed in significant quantities, and we speculate that the G-CSF swallowed by the fetus and neonate has local but not systemic effects. PMID- 12897303 TI - Prevention of pediatric overweight and obesity. AB - The dramatic increase in the prevalence of childhood overweight and its resultant comorbidities are associated with significant health and financial burdens, warranting strong and comprehensive prevention efforts. This statement proposes strategies for early identification of excessive weight gain by using body mass index, for dietary and physical activity interventions during health supervision encounters, and for advocacy and research. PMID- 12897304 TI - Prevention of medication errors in the pediatric inpatient setting. AB - Although medication errors in hospitals are common, medication errors that result in death or serious injury occur rarely. Even before the Institute of Medicine reported on medical errors in 1999, the American Academy of Pediatrics and its members had been committed to improving the health care system to provide the best and safest health care for infants, children, adolescents, and young adults. This commitment includes designing health care systems to prevent errors and emphasizing the pediatrician's role in this system. Human and device errors can lead to preventable morbidity and mortality. National and state legislative actions have heightened public awareness of these events. All involved persons, beginning with the physician and including every member of the health care team, must be better educated about and engaged in the several steps recommended to decrease these errors. The safe administration of medications to hospitalized infants and children requires additional specific safeguards that are above and beyond those for adult patients. Pediatricians should help hospitals develop effective programs for safely providing medications, reporting medication errors, and creating an environment of medication safety for all hospitalized pediatric patients. PMID- 12897305 TI - Prevention of drowning in infants, children, and adolescents. AB - Drowning is a leading cause of injury-related death in children. In 2000, more than 1400 US children younger than 20 years drowned. A number of strategies are available to prevent these tragedies. Pediatricians play an important role in prevention of drownings as educators and advocates. PMID- 12897306 TI - Prevention of drowning in infants, children, and adolescents. AB - Drowning is a leading cause of injury-related death in children. In 2000, more than 1400 US children younger than 20 years drowned. Most (91%) of these deaths were unintentional and were not related to boating. For each drowning death, it is estimated that at least 1 to 4 children suffer a serious nonfatal submersion event, many of which leave children with permanent disabilities. Environmental strategies, such as installation of 4-sided fences around swimming pools, and behavioral strategies, such as increased supervision of children while around water, are needed to prevent these tragedies. PMID- 12897307 TI - Caller satisfaction with after-hours telephone advice: nurse advice service versus on-call pediatricians. PMID- 12897308 TI - Meningitis--viral versus bacterial. PMID- 12897309 TI - The milk debate goes on and on and on! PMID- 12897310 TI - Successful 2 years of therapy. PMID- 12897311 TI - Caller satisfaction. PMID- 12897312 TI - Hospitalization for asthma: atopic, pulmonary function, and psychological correlates among participants in the Childhood Asthma Management Program. AB - BACKGROUND: Asthma in childhood has a significant impact on children and families, in part because of the frequent need for hospital-based care for acute exacerbations. Sensitization and exposure to inhalant allergens have been identified as risk factors for asthma hospitalization. OBJECTIVE: The Childhood Asthma Management Program (CAMP), comprised of 1041 children aged 5 to 12 with mild-to-moderate asthma, provides the opportunity to identify specific risk factors for prior hospitalization for asthma. METHODS: Data gathered during the screening period from CAMP were evaluated to elucidate differences between patients who had ever been hospitalized for asthma before enrollment in CAMP and those who had never been hospitalized. RESULTS: Univariate analyses indicated that prior hospitalization for asthma was associated with a younger age of asthma onset, longer duration of asthma, greater number of positive allergy skin tests, higher serum immunoglobulin E level, greater peripheral blood eosinophilia, greater recent inhaled corticosteroid use, greater airflow obstruction, greater airway hyperresponsiveness, and lower patient intelligence quotient (IQ). Gender, race, and family income did not differ between hospitalized and never hospitalized patients. The combination of both sensitization and exposure to high levels of dog allergen (Can f1) was associated with greater likelihood of prior hospitalization. Forward multivariate logistic regression analysis identified younger age of asthma onset, longer duration of asthma, recent use of asthma controller therapy, greater airflow obstruction, and lower patient IQ as significant risk factors for prior hospitalization when all risk factors identified by univariate analysis were included in the model. CONCLUSIONS: Children with mild-to-moderate asthma who had a past hospitalization for acute asthma had greater asthma severity, younger age of onset, and lower patient IQ at the time of entry into CAMP. They also had more markers of atopy than children without prior hospitalization, although atopy was not associated with prior hospitalization on multivariate analysis. Although we have identified these risk factors in a retrospective manner, one can speculate that the persistence of these features should alert the clinician to closely follow abnormalities on pulmonary function tests and general features of atopy to potentially identify patients at risk for future hospitalization. PMID- 12897313 TI - Long-term neuropsychologic and health outcomes of children with facial nerve palsy attributable to Lyme disease. AB - BACKGROUND: There is little information about the long-term outcomes of children with facial nerve palsy attributable to Lyme disease, a group putatively at high risk for poor neurologic outcomes. OBJECTIVE: The purpose of this study is to assess the long-term neuropsychologic and health outcomes of children with facial nerve palsy attributable to Lyme disease. METHODS: We conducted a matched cross sectional study of children with facial nerve palsy in Connecticut who met the Centers for Disease Control and Prevention national surveillance case definition for Lyme disease. We identified children with facial nerve palsy attributable to Lyme disease from population-based data for the state of Connecticut from 1984 to 1991 as well as from selected clinical practices from 1984 to 1998. For each case, 2 controls without Lyme disease, matched by age, were selected by sequential digit dialing technique. Both patients and controls (or their parents) were interviewed using structured questionnaires. Outcomes included self/parents' reports both of symptoms and of abilities to perform normal daily activities since the diagnosis was made (or for an equivalent period of time for controls). In addition, the patients with facial nerve palsy attributable to Lyme disease were evaluated with a battery of neuropsychologic tests. RESULTS: Of the 43 patients, 30% were female and 93% were white. Median age at diagnosis was 8 years (range: 2-18 years). Mean time to follow-up was 49 months (range: 7-161 months). Of the patients, 16% had been treated parenterally with ceftriaxone and 84% had been treated orally with either doxycycline or amoxicillin. Overall, 79% believed they were cured of Lyme disease, 9% believed they were not cured, and 12% did not know. The proportions of patients and of matched controls that reported increased problems with normal daily activities (eg, exercise, academic performance, word recall, sleeping) were similar. Patients with facial nerve palsy attributable to Lyme disease were more likely than matched controls to report symptoms of neck pain, changes in behavior, pains in joints or muscles, numbness or funny sensations in nerves, and problems with memory, but three quarters of them did not attribute these problems to Lyme disease. Twenty patients agreed to undergo neuropsychologic testing; all had average to above average scores on tests of IQ (eg, mean IQ was 110.2), memory, learning, attention, concentration, math, reading and spelling performance, executive functioning, abstract thinking, sequential processing, visuospatial scanning, and motor speed. There were no apparent differences in outcomes between children who received oral versus parenteral treatment. CONCLUSIONS: The neuropsychologic and health outcomes of children with facial nerve palsy attributable to Lyme disease 7 to 161 months earlier are comparable to those who did not have Lyme disease. PMID- 12897314 TI - A retrospective cohort study of the association of varicella vaccine failure with asthma, steroid use, age at vaccination, and measles-mumps-rubella vaccination. AB - OBJECTIVE: Varicella breakthrough, the occurrence of varicella disease >42 days after vaccination, is indicative of vaccination failure. A sevenfold increased risk of breakthrough among vaccinated children with asthma was observed in a 1996 varicella outbreak in a child care center. More recent outbreak investigations have also identified age at vaccination as a potential risk factor for breakthrough. We assessed the association of varicella breakthrough with asthma, steroids, age at varicella vaccination, and timing of measles-mumps-rubella (MMR) vaccination. METHODS: We performed a retrospective cohort study among children born after 1993 and followed up through 1999 at 2 health maintenance organizations ([HMOs] A and B) in the United States. Information was obtained from automated vaccination, clinic, hospital discharge, and pharmacy records. RESULTS: We identified 268 and 97 breakthrough cases among 80 584 and 8181 children vaccinated against varicella at HMOs A and B, respectively. Varicella breakthrough was not associated with asthma, inhaled steroids prescribed at any time, and oral steroids prescribed before vaccination. An increased risk of varicella breakthrough was found in the 3 months immediately after prescription for oral steroids at HMO A (adjusted relative risk [aRR]: 2.4; 95% confidence interval [CI]: 1.3-4.4) and HMO B (aRR: 2.8; 95% CI: 1.0-7.8), when varicella vaccine was given before 15 months of age at HMO A (aRR: 1.4; 95% CI: 1.1-1.9), and when varicella vaccination followed MMR vaccine within 28 days at HMO A (aRR: 3.1; 95% CI: 1.5-6.4). CONCLUSIONS: Varicella vaccine failure in children was not associated with asthma or the use of inhaled steroids, but with the use of oral steroids. Administration of varicella vaccine before the age of 15 months may be associated with a slightly increased risk of breakthrough disease. As currently recommended, varicella vaccination should not be administered for 28 days after MMR vaccination. PMID- 12897315 TI - Vancomycin use in hospitalized pediatric patients. AB - OBJECTIVES: To assess vancomycin utilization at children's hospitals, to determine risk factors for vancomycin use and length of therapy, and to facilitate adapting recommendations to optimize vancomycin prescribing practices in pediatric patients. METHODS: Two surveys were conducted at Pediatric Prevention Network hospitals. The first (Survey I) evaluated vancomycin control programs. The second (Survey II) prospectively reviewed individual patient records. Each hospital was asked to complete questionnaires on 25 consecutive patients or all patients for whom vancomycin was prescribed during a 1-month period. RESULTS: In Survey I, 55 of 65 (85%) hospitals reported their vancomycin control policies. Three quarters had specific policies in place to restrict vancomycin use. One half had at least 3 vancomycin restriction measures. In Survey II, personnel at 22 hospitals reviewed 416 vancomycin courses, with 2 to 25 (median = 12) patients tracked per hospital. Eighty-two percent of the vancomycin prescribed was for treatment of neonatal sepsis, fever/neutropenia, fever of unknown origin, positive blood culture, pneumonia, or meningitis. In an additional 6% (26/416), vancomycin was prescribed for patients with beta-lactam allergies and in 13% (56/416) for prophylaxis. Median duration of prophylaxis was 2 days (range: 1-15 days). Almost half (196, 47%) of the patients who received vancomycin were in intensive care units; 27% of the vancomycin courses were initiated by neonatologists and 19% by hematologists/oncologists. The predominant risk factor at the time of vancomycin initiation was the presence of vascular catheters (322, 77%); other host factors included cancer chemotherapy (55, 13%), transplant (30, 7%), shock (24, 6%), other immunosuppressant therapy (17, 4%), or hyposplenic state (2, <1%). Other clinical considerations were severity of illness (96, 23%), uncertainty about diagnosis (51, 12%), patient not responding to current antibiotic therapy (40, 10%), or implant infection (13, 3%). When vancomycin was initiated, blood cultures were positive in 85 patients (20%); cultures from other sites were positive in 45 (11%), and Gram stains of body fluids were positive in 37 (9%). In 29 (7%) patients, organisms sensitive only to vancomycin were isolated before vancomycin initiation. Reasons for discontinuing vancomycin included: therapeutic course completed (125, 30%), negative cultures (106, 25%), alternative antibiotics initiated (75, 18%), illness resolved (14, 3%), or patient expired (13, 3%). Final results of blood culture isolates resistant to beta-lactam antibiotics included 48 coagulase-negative staphylococcus, 5 Staphylococcus aureus, and 10 other species. CONCLUSIONS: At children's hospitals, vancomycin is initiated for therapy in patients who have vascular catheters and compromised host factors. Only 7% had laboratory-confirmed beta-lactam-resistant organisms isolated at the time vancomycin was prescribed. Efforts to modify empiric vancomycin use in children's hospitals should be targeted at intensivists, neonatologists, and hematologists. Initiatives to decrease length of therapy by decreasing the number of surgical prophylaxis doses and days of therapy before laboratory results may decrease vancomycin exposure. PMID- 12897316 TI - Health of children in title XXI: should we worry? AB - BACKGROUND: Capitation rates for the State Children's Health Insurance Program (SCHIP) funded under Title XXI of the Social Security Act were based on assumptions about the health care needs of children enrolled in this program. It has been suggested that parents are selective in enrolling children who are, in their opinion, most likely to need care, and that families who do not view their children as needing such care are more likely to ignore opportunities to seek or to maintain enrollment in SCHIP insurance. Thus, there have been concerns that enrollees might have more health conditions than a general population of children. OBJECTIVE: The purpose of this study was to test the hypothesis that children in Title XXI have more ongoing health conditions than expected by comparing health status data from enrollees in 1 state SCHIP program to a nationally representative sample of children in the United States. METHODS: This study used statewide data obtained in a survey of Florida SCHIP program enrollees and national data obtained on a subset of the children who were assessed in the 1994 National Health Interview Survey (NHIS). We examined health and demographic data collected by means of a structured telephone survey from parents for a random sample of 2432 children 2 to 18 years old who participated in the Florida Healthy Kids Program during the time period of October 1, 1997 through September 30, 1998. We compared these data to information on a national sample of all 26 845 children in the same age range whose health was assessed by the 1994 NHIS, and to a subset of the 6460 children in the NHIS sample whose family income met the eligibility criteria for SCHIP. To do this, we eliminated those children who were receiving Medicaid and those children whose household income levels were known to be above the eligibility level for SCHIP. We also excluded children whose household income was not reported. Thus, effectively this subsample contained non-Medicaid children, whose family incomes were below 185% of the federal poverty level. In Florida, a parent interview conducted by telephone included the Questionnaire for Identifying Children With Chronic Conditions (QuICCC), and their responses were used to determine if the child had any chronic health conditions. The QuICCC is a validated questionnaire containing 39-item sequences that ask about consequences of health conditions in children. It not only provides an overall classification of whether the child has a chronic condition, but also yields information about the consequences affecting the child within 3 condition-related domains: 1) functional limitations; 2) dependency on compensatory mechanisms or assistance; and 3) service need or use above and beyond routine care for age. The 1994 NHIS Core Interview and Disability Supplement contained a series of questions about children's functioning and service use that simulated the QuICCC, and we applied a previously published algorithm for determining the presence of a chronic condition using these items. Both data sets included comparable information on parental ratings of children's health status (excellent, very good, good, fair, poor), and on school absences, bed days, and restricted activity days in the previous 2 weeks. RESULTS: Children in the Florida SCHIP program were more than twice as likely to have chronic health conditions than similarly aged children in the general population or children in the income restricted national subsample (31% vs 15.9% and 14.6%, respectively) and there also were more SCHIP children with school absences (29% vs 18% and 16.7%, respectively). In contrast, the rating of overall health of SCHIP children was not poorer according to their parents, and they did not have more activity restrictions. Children in Florida SCHIP who had conditions were more likely to experience related consequences within each of 3 domains, and they were nearly 3 times as likely to have all 3 types of consequences (7% in Florida SCHIP sample vs 2.8% and 1.7% in the full national sample and in the income matched subsamome-matched subsample). However, the Florida SCHIP enrollees differed from the age and income-matched national sample in terms of the proportion of Hispanics. Thus, to verify these findings, we weighted the SCHIP sample to match the racial and ethnic proportions found in the national sample and repeated the analyses. The findings were robust, and there was no change in the percentage of children with special health care needs after such weighting. CONCLUSIONS: Overall, the results of these analyses support the notion of adverse selection and retention in the SCHIP program. This is unlikely to be the result of aggressive marketing in enrollment sites that serve children with more medical problems, as Florida health care providers rank third as a source of information about the Title XXI program after family and friends and the schools. In addition, Florida has active outreach and single-page application process for Medicaid and an aggressive program to move children to Title V, which also should minimize the numbers of children with special health care needs enrolled in SCHIP. Nevertheless, these findings suggest that the children being enrolled in Florida's SCHIP program are not the largely healthy population that was envisioned. If replicated in other SCHIP programs, these findings raise questions about the basic underlying assumptions concerning the health of potential enrollees and could have implications for the long-term fiscal viability of the program. PMID- 12897317 TI - A family-based approach to the prevention of depressive symptoms in children at risk: evidence of parental and child change. AB - OBJECTIVE: Depression in parents is a prevalent and impairing illness that is encountered frequently in medical practice. Children of depressed parents are at risk for psychopathology and other difficulties. A series of recent national reports have recommended the development of prevention efforts targeting children of depressed parents. Yet, to date, few controlled prevention studies of depression in children and adolescents have been conducted. In this study, we report the evaluation of 2 preventive intervention strategies that target children living in homes with depressed parents. Both are public health approaches that were designed to be used by a wide range of practitioners from a variety of disciplines, including pediatricians, internists, school counselors, nurses, and mental health practitioners. We adopted a developmental perspective and intervened with families when children were entering the age of highest risk for depression onset (ie, adolescence). We chose a family-based approach to prevention and sought to reduce risk factors and enhance protective factors for early adolescents by increasing positive interactions between parents and children, and by increasing understanding of the illness for everyone in the family. Our prevention approaches were designed to provide information about mood disorders to parents, to equip parents with the skills they need to communicate information to their children, and to open a dialogue with their children about the effects of parental depression. We hypothesized that participation in these prevention programs would result in parental change in child-related behaviors and attitudes about depression and its impact on the family. In addition, we hypothesized that this parental change would produce change in children's self understanding, and in children's depressive symptomatology. METHODS: We conducted a large-scale efficacy trial of 2 manual-based preventive intervention programs that were designed to be used widely in public health settings. These interventions target the relatively healthy children (ages 8-15) of parents with mood disorder. Ninety-three families (88.5% of our initial sample), including 121 children, participated in this study through the fourth assessment point. These families were assigned randomly to either a lecture or a clinician-facilitated intervention. Both interventions were specified in manuals. The lecture condition consisted of 2 separate meetings delivered in a group format without children present. The clinician-facilitated condition consisted of 6 to 11 sessions, including separate meetings with parents and children, and a family meeting in which the parents led a discussion of the illness and of positive steps that can be taken to promote healthy functioning in the children. In addition, telephone contacts or refresher meetings were conducted at 6- to 9-month intervals. In both conditions, psychoeducational material about mood disorders, risk, and resilience was presented and efforts were made to decrease feelings of guilt and blame in children. Parents were helped to build resilience in their children through encouraging their friendships, their success outside of the home, and their understanding of parental illness and of themselves. In addition, in the clinician-facilitated condition, efforts were made to link the psychoeducational material presented to the family's own unique illness experience. To address directly how their lives had changed, all family members in both conditions were assessed for psychopathology and for overall functioning at intake, and for psychopathology, functioning, and response to intervention immediately postintervention, approximately 1 year postintervention, and again approximately 2.5 years postintervention. RESULTS: We examined the outcomes of child understanding and internalizing symptomatology, and a number of predictor variables, using repeated measures analyses with generalized estimating equations. We found that parents in both conditions reported significant change in child-related behaviors and and attitudes, and that the amount of change reported increased over time from time 3 to time 4 (chi2(1) = 18.1). Moreover, relative to parents in the lecture program (mean number of changes = 6.3), parents in the clinician-facilitated program reported more change in child related behaviors and attitudes (mean number of changes = 9.8). Children in both conditions reported increased understanding of parental illness attributable to participation in our intervention programs. There was a positive association between the amount of change children reported in their understanding of parental illness and the number of changes couples reported in child-related behaviors/attitudes (chi2(1) = 37.3; ie, parents who had changed the most in response to intervention had children who also changed the most). Finally, internalizing scores for all children decreased with increased time since intervention (chi2(1) = 7.3). In addition, females had higher internalizing scores than males (chi2(1) = 5.3). There was no significant effect of group on children's change in internalizing symptomatology (chi2(1) = 0.2). CONCLUSIONS: We enrolled families with relatively healthy children, administered carefully designed preventive interventions that are manual-based and relatively brief, and found that these programs do have long-standing positive effects in how families problem solve around parental illness. Our results show significant benefits from both interventions. Moreover, changes in parents' perceptions translated directly into changes in children's own understanding of parental illness. Parental behavior and attitude changes and their connection to child changes in understanding identify an important mediating variable: family change. By increasing children's understanding of parental mood disorder, our interventions were found to promote resilience-related qualities in these children at risk. This presentation represents the first and only longitudinal primary prevention study of relatively healthy children at risk for psychopathology attributable to parental mood disorder and demonstrates a significant reduction in risk factors and increase in protective factors in these families over a long time interval- 2(1/2) years. Our results provide support for a family-based approach to preventive intervention. PMID- 12897318 TI - Vitamin D, cod-liver oil, sunlight, and rickets: a historical perspective. AB - Rickets, a disease of vitamin D deficiency, is rarely confronted by the practicing pediatrician in the United States today. At the turn of the 20th century, rickets was rampant among the poor children living in the industrialized and polluted northern cities of the United States. With the discovery of vitamin D and the delineation of the anti-rachitic properties of cod-liver oil by the 1930s, it became possible to not only treat but also eradicate rickets in the United States. Rickets was a common disease in 17th century England. Frances Glisson's treatise on rickets published in 1650, a glorious contribution to English medicine, described the clinical and anatomic features of rickets in great detail. The exact etiology of rickets had been elusive until the 1920s. During the Glissonian era, rickets was a mysterious disease. By the late 19th and early 20th century, faulty diet or faulty environment (poor hygiene, lack of fresh air and sunshine) or lack of exercise was implicated in its etiology. Animal experiments, appreciation of folklore advocating the benefits of cod-liver oil, and the geographical association of rickets to lack of sunshine were all relevant factors in the advancement of knowledge in the conquest of this malady. In this article, the history of rickets pertaining to the discovery of vitamin D, cod-liver oil, and sunlight is reviewed. PMID- 12897319 TI - A cohort study of health effects of human T-cell lymphotropic virus type I infection in Jamaican children. AB - OBJECTIVE: Human T-cell lymphotropic virus type I (HTLV-I) infection in childhood is believed to play an important role in risk for adult T-cell leukemia/lymphoma. Although HTLV-I is known to be associated with infective dermatitis in childhood, other HTLV-I-associated morbidity in children has not been well studied. We sought to determine the HTLV-I-associated health effects in Jamaican children. METHODS: We compared incidence rates of several health outcomes in 28 HTLV-I infected and 280 uninfected children clinically followed from age 6 weeks to a maximum of 10 years. Cox proportional hazards regression analysis was used to analyze these prospectively collected data, adjusting for confounding effects of other variables as necessary. RESULTS: HTLV-I-infected children had significantly higher incidence rates of seborrheic dermatitis (rate ratio [RR] = 4.8, 95% confidence interval [CI] = 1.9-12.5), eczema (RR = 3.1, CI = 1.2-7.9) and persistent hyperreflexia (RR = 3.7, CI = 1.6-8.2). Additionally, HTLV-I infected children had increased rates of severe anemia (RR = 2.5, CI = 0.8-7.9) and abnormal lymphocytes (RR = 2.4, CI = 0.8-7.6) that were of borderline statistical significance. CONCLUSIONS: Our study suggests that HTLV-I-associated skin diseases of childhood may include seborrheic dermatitis and eczema. Additionally, these data suggest that persistent hyperreflexia of the lower limbs may be an early sign of HTLV-I-associated neurologic involvement in children. Expansion and continued clinical observation of this cohort would be valuable. PMID- 12897320 TI - Determinants of health care use by children in rural western North Carolina: results from the Mountain Accessibility Project. AB - OBJECTIVE: To assess determinants of health care visits among children in a 12 county region of western North Carolina representative of rural areas in the United States. METHODS: Households were randomly selected for surveys of household characteristics, health status, and health care use. Surveys were conducted June 1999 to January 2000 and were stratified for children younger than 5 years and 5 years and older. The number of health care visits in the year before the survey was used as the outcome measure. Weighted mean visits and associations of family demographic and child health variables with the number of visits were determined by ratio and multivariate survey regression methods. RESULTS: Among children who lived in rural Appalachian regions of North Carolina in 1999, 90% had either public or private insurance coverage. The mean number of visits per child was 5.7 (median: 2.6), and in each age group the number of visits in the previous year exceeded the recommended number of well-child visits. There were no apparent geographic access barriers to care in this population, in that increased distances to provider sites did not result in declining numbers of visits. For children younger than 5 years, the primary determinants of health care use during the previous year were age, insurance status, and household income. Infants had more visits than older, preschool children, and those with household incomes >40 000 dollars per year had 76% more visits than those with incomes <20 000 dollars per year. Children with public insurance, exclusively Medicaid in this population, had almost 4 times as many visits as uninsured children. Among the children and adolescents 5 through 17 years of age, health insurance status, household income, pain during the past month, and race were the primary determinants of health care use during the previous year. Those with public health insurance had 6 times more health care visits than uninsured children. Household incomes >40 000 dollars per year were associated with 2.5 fold increased health care visits, and those with household incomes between 20 000 dollars and 40 000 dollars per year had 2-fold increased health care visits, compared with those with household incomes <20 000 dollars per year. White children had almost twice as many visits in the past year as black children in this age group. Pain experienced during the past month, as perceived by the parent, also predicted the number of visits in the older age group. CONCLUSIONS: This rural population seems to have reasonably good access to care overall. The key determinants of health care use among these rural children were similar to those found in urban and other populations in the United States and likely are universal: health insurance coverage, household income, and parent perceptions of their child's pain. As in other populations, programs in rural areas that strengthen health insurance coverage and reduce poverty will have a direct impact on child health. Differential use of health care among white and black children, especially those 5 years and older, merits additional explanation. PMID- 12897321 TI - Congenital cytomegalovirus infection in twin pregnancies: viral load in the amniotic fluid and pregnancy outcome. AB - Human cytomegalovirus (CMV) is the most common cause of viral intrauterine infection and fetal damage largely attributable to maternal primary infection. Most cases of congenital CMV infection in twins reported in the literature involved only 1 twin. We assessed the validity of polymerase chain reaction (PCR) and quantitative PCR on amniotic fluid (AF), at 21 to 22 weeks' gestation and at least 6 to 8 weeks after seroconversion, to predict the outcome of newborns in twin pregnancies. Two pregnant women with twin pregnancies and 1 woman with a triple pregnancy with primary CMV infection defined by the presence of immunoglobulin (Ig) M and low IgG avidity and/or by the presence of clinical symptoms and abnormal liver enzyme values were evaluated. CMV infection was found in 6 fetuses/newborns, 3 of whom were symptomatic. In the first twin pregnancy with diamniotic-dichorionic separate placentas, CMV symptomatic infection of the female twin was demonstrated by positive virus isolation and high viral load in AF. The male fetus was not infected as demonstrated by negative CMV culture and DNA detection in AF. In the triple pregnancy, the woman had a placenta with 2 monozygotic twins (females) and a separate placenta with a heterozygotic twin (male). The quantitative PCR results were 10(3) genome equivalents (GE)/mL of females AF and 1.9 x 10(5) GE/mL of male AF. Both female twins were asymptomatic at birth, whereas the male presented petechiae, thrombocytopenia, and cerebral ventriculomegaly. In the last twin pregnancy with fused dichorionic placentas, congenital CMV infection of both twins was diagnosed at birth in contrast with prenatal diagnosis. At time of amniocentesis, the left side twin was not infected as shown by negative results of CMV culture and DNA detection in the AF. CMV infection of the right side twin was demonstrated by positive CMV DNA detection with a CMV DNA load of 4.9 x 10(4) GE/mL and positive virus isolation in the AF. The morphologic and histologic examinations of the placentas strongly supported a prenatal horizontal acquisition of CMV infection. These twin pregnancies showed a marked difference in the quantity of virus load documented by the prenatal diagnosis suggesting that twin fetuses may react differently to primary maternal infection despite being exposed to the same maternal influences. A high viral load is correlated with congenital CMV infections symptomatic at birth. In such cases, with fetal infection of only 1 twin (at amniocentesis) and fusion of placentas, fetal outcome of both twins needs to be evaluated for the possibility of viral transfer from one fetus to the other. PMID- 12897322 TI - Congenital methemoglobinemia: a rare cause of cyanosis in the newborn--a case report. AB - Cyanosis is a physical finding that can occur at any age but presents the greatest challenge when it occurs in the newborn. The cause is multiple, and it usually represents an ominous sign, especially when it occurs in association with neonatal sepsis, cyanotic congenital heart disease, and airway abnormalities. Cyanosis caused by abnormal forms of hemoglobin can also be life-threatening, and early recognition is mandatory to prevent unnecessary investigations and delay in management. Abnormal hemoglobin, such as hemoglobin M, is traditionally discovered by electrophoresis, so the newborn screen, which is mandatory in several states, is a useful tool for the diagnosis. Although acquired methemoglobinemia, caused by environmental oxidizing agents, is common, congenital deficiency of the innate reducing enzyme is so rare that only a few cases are documented in the medical literature around the world. We present a neonate with cyanosis as a result of congenital deficiency of the reduced nicotinamide adenine dinucleotide-cytochrome b5 reductase enzyme. This infant was found to be blue at a routine newborn follow-up visit. Sepsis, structural congenital heart disease, prenatal administration, and ingestion of oxidant dyes were excluded as a cause of the cyanosis by history and appropriate tests. Chocolate discoloration of arterial blood provided a clue to the diagnosis. A normal newborn screen and hemoglobin electrophoresis made the diagnosis of hemoglobin M unlikely as the cause of the methemoglobinemia (Hb A 59.4%, A2 1.8%, and F 38.8%). Red blood cell enzyme activity and DNA analysis revealed a homozygous form of the cytochrome b5 reductase enzyme deficiency. He responded very well to daily methylene blue and ascorbic acid administration, and he has normal growth and developmental parameters, although he shows an exaggerated increase in his methemoglobin level with minor oxidant stress such as diarrhea. PMID- 12897323 TI - Cerebral embolism from atrial myxoma in pediatric patients. AB - We describe 2 children with cerebrovascular events caused by emboli from left atrial myxomas and review 7 other pediatric cases from the literature. Transient cutaneous eruptions involving the extremities preceded the cerebrovascular events and were most likely attributable to fragmentation of the atrial tumor with peripheral embolization. Our first case demonstrates the more common presentation with acute hemiplegia caused by cerebral infarction and the second case a transient ischemic attack manifested by more subtle features as a result of involvement of the vertebrobasilar circulation. Neither child had a history or other signs of cardiac disease. Atrial myxoma should be considered in the differential diagnosis when children present with neurologic symptoms or with signs of embolization, because surgical removal of the tumor is critical and may be curative. PMID- 12897324 TI - Uninsured children: how we count matters. AB - BACKGROUND: Because children uninsured for less than a full year are often reported as insured, they receive less attention in health policy debates than do the full-year uninsured and are underrecognized as potential users of public insurance programs. OBJECTIVE: The purpose of this study is to assess the impact on estimates of how many US children are uninsured when alternatives to the full year uninsured definition are used. METHODS: Monthly health insurance coverage data collected from children through age 18 in the 1999 Medical Expenditure Panel Survey were analyzed to estimate prevalence of health insurance gaps among children in terms of the size of part-year and full-year uninsured child population, duration of uninsured gaps, and aggregate uninsured spells. RESULTS: Although 6.6 million ([M] 8.4%) children in the United States were uninsured throughout 1999, an additional 11.4M (14.4%) were uninsured for part of the year. Part-year uninsured gaps accounted for 41.7% of a total of 130M months of missing coverage experienced by all children. CONCLUSIONS: Different definitions and measures of who are uninsured can project radically different pictures of the magnitude of the problem. As this study shows, including the part-year uninsured more than doubled the estimated uninsured child population for 1999, and increased the estimated aggregate uninsured months by 71%. As potential users of public coverage, children who have no insurance for part of the year should be included when evaluating challenges to and accomplishments by the State Children's Health Insurance Program and other public programs. Their significant numbers and the potential burden they place on the health care delivery system argue for them to be counted and for the causes and consequences of short-term uninsured spells to be better understood. PMID- 12897325 TI - State of your journal: a mid-year report of unprecedented progress. PMID- 12897327 TI - FDA drug approval summary: gefitinib (ZD1839) (Iressa) tablets. AB - On May 5, 2003, gefitinib (Iressa), ZD1839) 250-mg tablets received accelerated approval by the U.S. Food and Drug Administration as monotherapy treatment for patients with locally advanced or metastatic non-small cell lung cancer (NSCLC) after failure of both platinum-based and docetaxel chemotherapies. Information provided in this summary includes efficacy and safety results of relevant clinical trials. Effectiveness was demonstrated in a randomized, double-blind, phase II, multicenter trial comparing two oral doses of gefitinib (250 mg/day versus 500 mg/day). Two hundred sixteen patients were enrolled. The 142 patients who were refractory to or intolerant of a platinum and docetaxel comprised the evaluable population for the efficacy analysis. A partial tumor response occurred in 14% (9 of 66) of patients receiving gefitinib 250 mg/day and in 8% (6 of 76) of patients receiving gefitinib 500 mg/day. The overall objective response rate for both doses combined was 10.6% (15 of 142 patients) (95% confidence interval 6.0%-16.8%). Responses were more frequent in females and in nonsmokers. The median duration of response was 7.0 months (range 4.6-18.6+ months). Other submitted data included the results of two large trials conducted in chemotherapy naive, stage III and IV NSCLC patients. Patients were randomized to receive gefitinib (250 mg or 500 mg daily) or placebo, in combination with either gemcitabine plus cisplatin (n = 1,093) or carboplatin plus paclitaxel (n = 1,037). Results from those studies showed no benefit (response rate, time to progression, or survival) from adding gefitinib to chemotherapy. Consequently, gefinitib is only recommended for use as monotherapy. Common adverse events associated with gefitinib treatment included diarrhea, rash, acne, dry skin, nausea, and vomiting. Most toxicities were Common Toxicity Criteria grade 1 or 2. Interstitial lung disease (ILD) has been observed in patients receiving gefitinib. Worldwide, the incidence of ILD is about 1% (2% in the Japanese postmarketing experience and about 0.3% in a U.S. expanded access program). Approximately one-third of the cases were fatal. Physicians should promptly evaluate new or worsening pulmonary symptoms. If ILD is confirmed, appropriate management includes discontinuation of gefitinib. Gefitinib was approved under accelerated approval regulations on the basis of a surrogate end point response rate. No controlled gefitinib trials, to date, demonstrate a clinical benefit, such as improvement in disease-related symptoms or greater survival. Accelerated approval regulations require the sponsor to conduct further studies to verify that gefitinib therapy produces such a benefit. PMID- 12897328 TI - The Her-2/neu gene and protein in breast cancer 2003: biomarker and target of therapy. AB - The HER-2/neu oncogene encodes a transmembrane tyrosine kinase receptor with extensive homology to the epidermal growth factor receptor. In this review, the association of HER-2/neu gene and protein abnormalities with prognosis and response to therapy with trastuzumab and to other therapies in breast cancer is presented. By considering a series of 80 published studies encompassing more than 25,000 patients, the relative advantages and disadvantages of Southern blotting, polymerase chain reaction amplification, and fluorescence in situ hybridization assays designed to detect HER-2/neu gene amplification are compared with HER 2/neu protein overexpression assays performed by immunohistochemical techniques applied to frozen and paraffin-embedded tissues and enzyme immunoassays performed on tumor cytosols. The significance of HER-2/neu overexpression in ductal carcinoma in situ and the HER-2/neu status in uncommon female breast conditions and male breast cancer are also considered. The role of HER-2/neu testing for the prediction of response to trastuzumab therapy in breast cancer is presented as well as its potential impact on responses to standard and newer hormonal therapies, cytotoxic chemotherapy, and radiation. The review also evaluates the status of serum-based testing for circulating HER-2/neu receptor protein and its ability to predict disease outcome and therapy response. PMID- 12897329 TI - Behavioral risk factors in breast cancer: can risk be modified? AB - The International Agency for Research on Cancer estimates that 25% of breast cancer cases worldwide are due to overweight/obesity and a sedentary lifestyle. The preponderance of epidemiologic studies indicates that women who engage in 3-4 hours per week of moderate to vigorous levels of exercise have a 30%-40% lower risk for breast cancer than sedentary women. Women who are overweight or obese have a 50%-250% greater risk for postmenopausal breast cancer. Alcohol use, even at moderate levels (two drinks per day) increases risk for both premenopausal and postmenopausal breast cancer. Certain dietary patterns, such as high fat, low vegetables/fruits, low fiber, and high simple carbohydrates, may increase risk, but definitive data are lacking. These lifestyle factors are likely associated with breast cancer etiology through hormonal mechanisms. The worldwide trends of increasing overweight and obesity and decreasing physical activity may lead to an increasing incidence of breast cancer unless other means of risk reduction counteract these effects. Thus, adoption of lifestyle changes by individuals and populations may have a large impact on the future incidence of this disease. PMID- 12897330 TI - Advances in endocrine treatments for postmenopausal women with metastatic and early breast cancer. AB - For the past 25 years, the estrogen antagonist tamoxifen has been the hormonal treatment of choice for postmenopausal patients with hormone-sensitive metastatic and early breast cancer (EBC). However, tamoxifen is associated with certain tolerability and safety concerns. In addition, the hormonal options after progression are limited, and thus, alternative endocrine treatments have been developed. This review provides a synopsis of the newer alternatives in endocrine therapy of breast cancer: the aromatase inhibitors (AIs) and fulvestrant Faslodex), the estrogen receptor antagonist that downregulates estrogen and progesterone receptors and has no known agonist activity. The third-generation AIs, anastrozole and letrozole, have been shown to be as effective or more effective than megestrol acetate and tamoxifen as second- and first-line therapies for the treatment of postmenopausal patients with metastatic breast cancer, and exemestane has been approved for second-line use. Fulvestrant has been shown to be as effective as anastrozole as second-line therapy for metastatic breast cancer and has been approved in the U.S. for the treatment of postmenopausal women with hormone-receptor-positive metastatic breast cancer following progression on antiestrogen therapy. Anastrozole is the only AI with published clinical trial data and U.S. Food and Drug Administration approval for adjuvant therapy of postmenopausal women with EBC. The 'Arimidex,' Tamoxifen, Alone or in Combination (ATAC) trial, a double-blind, multicenter trial with 9,366 patients, compared tamoxifen with anastrozole, alone and in combination, as adjuvant endocrine treatment for postmenopausal patients with operable, invasive, EBC. The first analysis (at a median follow-up of 33.3 months) showed longer disease-free survival and, in general, better tolerability with anastrozole than with tamoxifen. This pattern was maintained at later analyses with a median follow-up of 47 months for efficacy and 37 months for safety and tolerability. Although longer follow-up is warranted, anastrozole appears to be a well documented choice of endocrine adjuvant therapy for postmenopausal women with hormone-responsive breast cancer. PMID- 12897331 TI - Health-related quality of life in female long-term colorectal cancer survivors. AB - Although the number of women who survive treatment for colorectal cancer is growing, little is known about the quality of life of long-term survivors. The purpose of analyses presented in this paper is to describe the overall health related quality of life of female long-term colorectal cancer survivors and the factors that may modify their levels of quality of life. A population-based sample of 726 Wisconsin women diagnosed with colorectal cancer from 1990-1991 was recontacted. Of the 443 women alive in 1999, 307 (69%) completed a follow-up questionnaire including the Medical Outcomes Study Short-Form 36 Health Status Survey, which is comprised of 36 items that generate nine domain scale scores and two summary scores: the Physical Component Summary score and the Mental Component Summary score. The mean follow-up was 9 years (range 7-11), and the mean age at follow-up was 72 years (range 43-85). The mean Physical Component Summary score was lower for participants with greater ages, greater numbers of comorbidities, and greater body masses at the time of follow-up. The mean Mental Component Summary score also was lower for participants with greater numbers of comorbidities. Differences associated with degree of comorbidity were observed for all eight domain scales. Female long-term survivors of colorectal cancer appear to report health-related quality of life comparable with that of similarly aged women in the general population. These data suggest that, over the long term, factors attributable to aging, body weight, and chronic medical conditions play more dominant roles in determining physical and mental health than factors related to the initial colorectal cancer diagnosis. PMID- 12897332 TI - Intensive concurrent chemoradiotherapy for head and neck cancer with 5 Fluorouracil- and hydroxyurea-based regimens: reversing a pattern of failure. AB - Combined modality programs that were developed over the past two decades demonstrated that the nonsurgical therapy of locoregionally advanced head and neck cancer is feasible and produces survival outcomes that are at least comparable with surgery. The systemic therapy of head and neck cancer has gained momentum in recent years. Several randomized studies have shown that the concurrent administration of chemotherapy and radiation therapy is superior to radiation therapy alone. In consecutive clinical studies since 1986, we have developed multiagent chemoradiotherapy regimens based on initial observations with the 5-fluorouracil (5-FU), hydroxyurea, and concomitant radiotherapy combination. Three consecutive multicenter phase II trials reported that the combination of 5-FU and hydroxyurea with either cisplatin or paclitaxel along with twice daily radiation therapy administered every other week is a highly effective regimen with local control rates that approach 90% and 3-year survival rates of approximately 60% in patients with stage IV disease. The vast majority of patients in these studies achieved anatomical organ preservation. A reversal of the historical pattern of failure was evident, with distant sites becoming the predominant site of failure in each trial. The paclitaxel-containing regimen was better tolerated than the cisplatin-containing regimen and was advanced to further clinical testing. The incorporation of induction chemotherapy may improve the results of treatment by targeting systemic micrometastatic disease. PMID- 12897333 TI - Docetaxel-based combined-modality chemoradiotherapy for locally advanced non small cell lung cancer. AB - The cytotoxic agent docetaxel not only has proven activity in non-small cell lung cancer-when used alone or in combination-but is also a potent radiosensitizer, and improved treatments are needed in all stages of this disease. In patients with locoregionally advanced (stage III) disease, docetaxel has shown efficacy with manageable toxicities when used alone or in combination with a platinum compound in a sequential manner before localized radical radiotherapy/surgery. Presently, therapeutic gains appear to be maximized by the use of concurrent chemotherapy and irradiation. This review focuses on research with combinations of docetaxel with either cisplatin or carboplatin and radiotherapy. Overall response and survival rates to date provide data worth pursuing. From phase I data, weekly docetaxel at 20 mg/m(2) plus cisplatin at 25 mg/m(2) or carboplatin to an area under the concentration time curve of 2 mg/ml*min with concurrent radiotherapy to 60 Gy over 6 weeks appear to be suitable for phase II trials. Predominant toxicities are esophagitis and neutropenia, but a low frequency of pulmonary toxicity is reported. Induction, concurrent, and consolidation docetaxel-based chemoradiotherapy in potentially resectable disease are all being investigated. Future research could include the investigation of computed tomography/ positron emission tomography-derived target volume radiotherapy, dose escalated therapy, and alternative fractionation schedules in combination with docetaxel-based cytotoxic chemotherapy. PMID- 12897334 TI - Thrombotic thrombocytopenic purpura associated with bone marrow metastasis and secondary myelofibrosis in cancer. AB - To examine the relationship between cancer and development of thrombotic microangiopathy (TM), the medical records of patients with known TM were examined in one institution from January 1981 to December 2002. Nine out of 93 patients with the established diagnosis of TM had active cancer. All nine of those patients had thrombotic thrombocytopenic purpura (TTP). Among those patients, two patients received chemotherapy prior to the development of TTP. Six of the seven patients who received no chemotherapy had extensive bone marrow metastasis and secondary myelofibrosis. There were two patients each with breast cancer, lung cancer, and stomach cancer. Severe anemia and thrombocytopenia with leukoerythroblastosis were prominent clinical features in all six patients. Four patients had neurological (mental) changes and three developed fever, but none had significant renal dysfunction. Upon establishing the diagnosis of TTP, four patients were treated with exchange plasmapheresis (EP) and two patients were treated with chemotherapy because there were no neurological changes. Three patients achieved complete remission of TTP, one with EP alone and two with chemotherapy. The one patient who achieved remission with EP alone was later treated with chemotherapy and survived for 2 1/2 years. The other three patients treated with EP alone died within 2 months after the diagnosis of TTP. Since TTP occurred in association with bone marrow metastasis and myelofibrosis in six patients among seven chemotherapy-untreated cancer patients, this marrow change was considered to be the possible cause of the development of TTP. It is recommended that all cancer patients with unexplained anemia and thrombocytopenia be evaluated for the coexistence of bone marrow metastasis and TTP. PMID- 12897335 TI - Venous thrombosis in cancer patients: insights from the FRONTLINE survey. AB - BACKGROUND: Venous thromboembolism (VTE) is a common complication in cancer patients and a significant cause of morbidity and mortality. However, little information is available on oncologists' perceptions of the risk of VTE and its management. The Fundamental Research in Oncology and Thrombosis (FRONTLINE) study is the first comprehensive global survey of thrombosis and cancer. The study was designed to collect data on the perceived risk and patterns of practice with regard to VTE in cancer patients undergoing surgical and medical management of their malignancy and to provide information on international and regional practice patterns, allowing for the design of research studies to answer the concerns of practicing clinicians. METHODS: Literature reviews were performed to provide a current evidence base against which to compare the findings, and a survey was developed under the guidance of an advisory board. A paper-based reply paid questionnaire was distributed globally between July and November 2001 to clinicians involved in cancer care and was made available on a dedicated website. FINDINGS: A total of 3,891 completed responses were available for analysis. Brain and pancreatic tumors were considered to carry a high risk for VTE, and 80% of respondents considered the use of central venous lines to be associated with a high risk of VTE. Marked differences were seen in the use of thromboprophylaxis for surgical and medical cancer patients, with over 50% of surgeons reporting that they initiated thromboprophylaxis routinely, while most medical oncologists reported using thromboprophylaxis in less than 5% of medical patients. Low molecular weight heparin (LMWH) was the most popular method of thromboprophylaxis employed in both surgical and medical patients and was more favored by European than U.S. clinicians. Some 20% of respondents reported using aspirin for prophylaxis, despite there being no reliable evidence for this agent as effective in prevention in this population. For the treatment of VTE, LMWH was again the most common initial treatment, although, for the long-term, oral anticoagulation therapy was widely adopted. Many patients were treated for VTE on an outpatient basis, and secondary prevention of VTE was typically continued for 3 to 6 months after an episode of deep vein thrombosis or for longer in the case of pulmonary embolism. INTERPRETATION: The results of the FRONTLINE survey demonstrate a need for guidelines to direct clinical practice in line with evidence-based data concerning cancer and VTE. Oncologists need to be educated regarding the true risks of VTE associated with certain cancers and on strategies for prevention and treatment to reduce the morbidity and mortality associated with VTE in all cancer patients. The study has also helped identify areas for future research. PMID- 12897336 TI - The molecular perspective: histone deacetylase. PMID- 12897337 TI - Congress shapes a prescription drug benefit. PMID- 12897339 TI - Salt and water: read the package insert. PMID- 12897340 TI - Endocrine and reproductive manifestations of sarcoidosis. PMID- 12897341 TI - The clinical features of takotsubo cardiomyopathy. AB - BACKGROUND: Cardiologists have recently recognized a reversible form of heart failure of unknown origin characterized by a takotsubo-shaped hypokinesis of the left ventricle on left ventriculography. AIM: To clarify the clinical features of this cardiomyopathy. DESIGN: Observational study. METHODS: Seven patients with reversible ventricular dysfunction were followed for 4.5 years. Clinical course, routine examinations, and cardiac catheterizations in each patient were documented. RESULTS: The cardiomyopathy developed in six elderly female and one male patients (mean age 75.3 years), all of whom had been exposed to stress. Cardiac enzymes did not significantly increase, but serum norepinephrine increased remarkably (1.19 ng/ml). Coronary angiography revealed normal coronary arteries. However, left ventriculography showed akinesis in the apical segments, together with hyperkinesis in the basal segments (a takotsubo shape). The abnormal kinesis normalized within 17.4 hospital days without any treatment in five patients, and with haemodynamic support for 3 days in the other two. Endocardial biopsies did not suggest any specific pathology. The cardiac events did not recur over a 1-4 year follow-up. DISCUSSION: Coronary vasospasm, myocarditis and other substantial diseases previously described were ruled out as the cause of takotsubo cardiomyopathy in our subjects. Prognosis was good without any form of treatment, provided that the patients survived the severe heart failure state. Catecholaminergic or adrenoceptor-hyperactive cardiomyopathy may be the cause of this cardiomyopathy. PMID- 12897342 TI - The interleukin-6 promoter polymorphism in Gaucher disease: a new modifier gene? AB - BACKGROUND: Individuals with Gaucher disease vary significantly with regard to degree of bone disease, but there are no predictive markers for severity of skeletal involvement. AIM: To determine the frequency of polymorphisms of interleukin-6 (IL-6) among patients with Gaucher disease, and the relationship to bone mineral density (BMD) and other markers of disease severity. DESIGN: Case control study. METHODS: Genotyping for the 174G --> C promoter polymorphism of IL 6 was performed in adult patients with Gaucher disease for whom there was concurrent bone mineral density (BMD) data and in healthy Ashkenazi Jewish controls. RESULTS: The prevalence of allelic variants (58% G/G, 36% G/C, and 6% C/C) was similar in Ashkenazi Jewish adults with Gaucher disease as in Ashkenazi Jewish controls, but significantly different (p < 0.05) from that reported among Caucasians. No statistically significant correlation was found between IL-6 genotypes and BMD or markers of severity of Gaucher disease. Patients with the C/C genotype had relatively mild Gaucher disease. DISCUSSION: The IL-6 polymorphisms appear to be distributed differently in Ashkenazi Jews than among other Caucasians. In Gaucher disease, the C/C genotype may be associated with a milder Gaucher phenotype, and may serve as a mitigating genetic modifier. PMID- 12897343 TI - Reducing delays in the diagnosis and treatment of Clostridium difficile diarrhoea. AB - BACKGROUND: The diagnosis of Clostridium difficile diarrhoea is often delayed. AIM: To assess and reduce delays in diagnosis and treatment. DESIGN: Two-part study: retrospective audit then prospective observational. METHODS: The retrospective study audited cases positive for C. difficile culture or toxin A between June 2000 and January 2001. Cases were reviewed regarding demographic characteristics and the timing from onset of symptoms to testing and treatment (n = 27). In the prospective study, efforts were made to increase the awareness of medical staff about C. difficile diarrhoea, and testing for C. difficile toxin A assay was substituted for stool culture and external toxin analysis. Data were prospectively analysed for demographic characteristics and time from onset of symptoms to testing and treatment (n = 24). RESULTS: There were no significant differences in age (76 vs. 78 years), male to female ratio (1:2 vs. 1:4), reason for admission, specialty responsible, or mortality rate (40% vs. 37%) during the presenting admission. Preceding antibiotic use, the combination of antibiotics used and subsequent treatment also did not differ between groups. The time from onset of diarrhoea to sampling was reduced from 4.7 days (range 3-30) to 0.8 days (range 0-5; p or approximately= 100, the disparity between our results and those of a Newtonian fluid of constant viscosity grows with a decreasing ratio of the DC to AC components of the pressure-gradient amplitude below 50%. In particular, for any purely oscillatory pressure-gradient (vanishing DC component), the Womersley solution is a particularly poor predictor of the amplitude and phase of wall shear rate for over half of the flow cycle. Under such circumstances, the analytical models presented here provide a simple and accurate means of estimating instantaneous wall shear rate, knowing only the pressure gradient and hematocrit. PMID- 12897418 TI - Noninvasive determination of perfused artery dimensions ex vivo using a pressure diameter relationship. AB - Based on the decisive effects of the hemodynamic and mechanical environments on the development and remodeling of arteries in vivo, several groups have cultured tissue-engineered vessels and excised vessels in various mechanically active perfusion systems. To facilitate the interpretation and design of such studies, accurate estimates of the applied forces and resulting stresses are required, which in turn require an accurate estimate of vessel dimensions. The measured pressure drop along the length of the vessel could be used to calculate the average inner diameter, but practical considerations, including the modest accuracy of many pressure transducers, limit this approach. Using nine porcine arteries harvested from pigs weighing between 25 and 100 kg, we show that when real-time measurements of the pressure drop and the outer diameter during a vasoactive event are fit to a theoretical model, offset errors in the pressure measurement can be compensated for and estimates of vessel wall transverse area with an average error of 4.1% (not exceeding 8.3%) are achieved. PMID- 12897419 TI - Rheological behavior and parameters of the in vitro model of lung surfactant systems: the role of the main phospholipid component. AB - The proposed in vitro model for studying the alveolar surface layer of the lungs enables one to investigate the surface intermolecular forces which influence the stability of the alveolus. The general role for the stability of the alveolus belongs to the phospholipids in the alveolar surfactant and predominantly to their main component dipalmitoylphosphatidylcholine (DPPC). The aim of the study was to investigate the rheological behavior of DPPC and exogenous surfactant preparations used in neonatal clinical practice. Data for the rheological behavior of the solutions of the commercially available surfactants, Infasurf, Exosurf and Survanta, as well as of DPPC (their main phospholipid component) at shear rates from 0.024 to 94.5 s(-1) under steady and transient flow conditions at 23 degrees C were obtained. Infasurf and Exosurf showed Newtonian rheological behavior, while Survanta revealed the shear-thinning behavior of a non-Newtonian pseudoplastic fluid. The rheological properties of aqueous solutions of DPPC containing 0.14 M NaCl at concentrations from 100 and 630 microg/ml of phospholipid (chosen from the dependence of the probability for bilayer film formation) were studied. Differences observed in the rheological properties of the exogenous surfactants were interpreted on the basis of their composition, the presence of other phospholipid components, certain additives and surfactant proteins, as well as the bulk structures formed from them. The relevance of the results for the delivery of exogenous surfactants and their spreading in replacement therapy is discussed. PMID- 12897420 TI - Lubricated squeezing flow: a useful method for measuring the viscoelastic properties of soft tissues. AB - Conducting experiments on very soft biological tissues can be difficult. Traditionally, unconfined compression and shear have been used. Here, an improved method of compression testing, lubricated squeezing flow is described. This gives a uniform compression along the squeezing axis and almost uniform equi-biaxial elongation at right angles to the squeezing axis, with minimal shear deformation due to the constant lubrication of the sample surfaces during testing. Sample results for porcine liver obtained using this method are described here. PMID- 12897421 TI - Viscoelastic deformation response of red blood cells under conditions of oscillating centrifugal field. AB - The red cell deformation under the conditions of oscillating centrifugal fields was studied. Experiments were carried out with a modified Cell-Elastometer operating in oscillating mode (0.02 to 0.30 Hz). Gravitational acceleration was sinusoidally modulated between 620 g and 2250 g. At low frequencies (below 0.08 Hz), native red cells followed the applied stress without delay. At 0.09 Hz and up, the cellular deformation was still periodical and included an additional perturbation due to intracellular movements. This perturbation was analysed and quantified. The influence of alterations on the erythrocyte membrane by diamide was analysed to verify the sensitivity of this method. On increasing the membrane stiffness with low concentrations of diamide, the response to oscillatory centrifugal stress was impaired characteristically in terms of amplitude deformation. Based on tangential and centrifugal accelerations, a physical model was developed that describes the basic observable changes on varying the oscillation frequency. From the data it can be concluded that viscoelastic properties of red cells can be analysed and quantified using oscillatory centrifugal accelerations. The described method can become a valid tool to differentiate between membrane alterations or intracellular viscous modifications. PMID- 12897422 TI - Increased resistance of neutrophils to deformation upon cooling and rate of recovery on rewarming. AB - Increase in the resistance to deformation of neutrophils upon exposure to the cold may impair their passage through microvessels. However, the potential for such rheological changes to cause prolonged microvascular obstruction in cooled tissue will depend on whether and at what rate the neutrophils recover on rewarming. We tested the ability of neutrophils to pass through micropore filters, and found that neutrophils cooled to 10 degrees C for 10-20 minutes could block either 5 microm or 8 microm pore filters. On return to 37 degrees C, flow resistance remained impaired briefly but recovered over about 5 minutes. The kinetics of changes in flow resistance in the cold and on rewarming were linked to kinetics of actin polymerisation during these periods. However, they were not closely linked to distortion of cell shape in the cold, which recovered only slowly with rewarming. The results suggest that while rigid neutrophils might occlude capillaries in cold tissue, mechanical obstruction should not be long lived on rewarming. Moreover, rigid neutrophils washed out of cold tissue should experience only temporary mechanical trapping in remote tissues. PMID- 12897423 TI - Oxidative signaling and glutathione synthesis. PMID- 12897424 TI - Cell survival and changes in gene expression in cells unable to synthesize glutathione. PMID- 12897425 TI - Role of glutathione in the regulation of liver metabolism. PMID- 12897426 TI - Glutathione transport in the endo/sarcoplasmic reticulum. AB - Glutathione transport through the endo/sarcoplasmic reticulum (ER/SR) membrane might play a role in the maintenance of the thiol redox potential difference between the lumen and the cytosol. The transport of glutathione (both GSH and glutathione disulfide, GSSG) is entirely different in the ER and SR membranes. The transport measurements based on either rapid filtration or light scattering techniques revealed that the SR membrane transports glutathione much faster than the hepatic ER membrane or microsomal membranes prepared from heart or brain. The fastest transport has been measured in the membrane of muscle terminal cisternae, which is enriched in ryanodine receptor type 1 (RyR1). All the studied membranes have been found to be equally impermeable to various hydrophilic substances of similar size to glutathione, thus the glutathione transport in muscle microsomes and terminal cysternae as well as the correlation between the rate of glutathione transport and the abundance of RyR1 are specific. In both muscle microsomes and terminal cysternae, glutathione influx can be either inhibited or activated by antagonists and agonists of the ryanodine receptor, respectively, while these agents do not influence the transport of other small permeant molecules. These findings strongly suggest that the ryanodine receptor channel activity is directly associated with glutathione transport activity in the skeletal muscle sarcoplasmic reticulum membrane. PMID- 12897427 TI - Role of ascorbate in oxidative protein folding. AB - Both in prokaryotic and eukaryotic cells, disulfide bond formation (oxidation and isomerization steps) are catalyzed exclusively in extracytoplasmic compartments. In eukaryotes, protein folding and disulfide bond formation are coupled processes that occur both co- and posttranslationally in the endoplasmic reticulum (ER), which is the main site of the synthesis and posttranslational modification of secretory and membrane proteins. The formation of a disulfide bond from the thiol groups of two cysteine residues requires the removal of two electrons, consequently, these bonds cannot form spontaneously; an oxidant is needed to accept the electrons. In aerobic conditions the ultimate electron acceptor is usually oxygen; however, oxygen itself is not effective in protein thiol oxidation. Therefore, a small molecular weight membrane permeable compound should be supposed for the transfer of electrons from the ER lumen. The aim of the present study was the investigation of the role of ascorbate/dehydroascorbate redox couple in oxidative folding of proteins. We demonstrated that ascorbate addition or its in situ synthesis from gulonolactone results in protein thiol (and/or glutathione; GSH) oxidation in rat liver microsomes. Since microsomal membrane is hardly permeable to ascorbate, the existence of a transport metabolon was hypothesized. Three components of the system have been described and partially characterized: (i) A microsomal metalloenzyme is responsible for ascorbate oxidation on the outer surface of the ER. Ascorbate oxidation results in ascorbate free radical and dehydroascorbate production. (ii) Facilitated diffusion of dehydroascorbate is present in microsomal vesicles. The transport is presumably mediated by a GLUT-type transporter. On the contrary, the previously hypothesized glutathione disulfide (GSSG) transport is practically absent, while GSH is transported with a moderate velocity. (iii) Protein disulfide isomerase catalyzes the reduction of dehydroascorbate in the ER lumen. Both GSH and protein thiols can be electron donors in the process. Intraluminal dehydroascorbate reduction and the consequent ascorbate accumulation strictly correlate with protein disulfide isomerase activity and protein thiol concentration. The concerted action of the three components of the system results in the intraluminal accumulation of ascorbate, protein disulfide and GSSG. In fact, intraluminal ascorbate and GSSG accumulation could be observed upon dehydroascorbate and GSH uptake. In conclusion, ascorbate is able to promote protein disulfide formation in an in vitro system. Further work is needed to justify its role in intact cellular and in vivo systems, as well as to explore the participation of other antioxidants (e.g. tocopherol, ubiquinone, and vitamin K) in the electron transfer chain responsible for oxidative protein folding in the ER. PMID- 12897428 TI - Cytophotometric investigations on oscillating thiol-disulfide equilibria and oxidized protein sulfur. PMID- 12897429 TI - Protection by pantothenic acid against apoptosis and cell damage by oxygen free radicals--the role of glutathione. PMID- 12897430 TI - Thiols as major determinants of the total antioxidant capacity. PMID- 12897431 TI - Enzymes of the thiol-dependent hydroperoxide metabolism in pathogens as potential drug targets. PMID- 12897432 TI - Is there a role of glutathione peroxidases in signaling and differentiation? PMID- 12897433 TI - Multidrug resistance-associated proteins: Export pumps for conjugates with glutathione, glucuronate or sulfate. AB - Many endogenous or xenobiotic lipophilic substances are eliminated from the cells by the sequence of oxidation, conjugation to an anionic group (glutathione, glucuronate or sulfate) and transport across the plasma membrane into the extracellular space. The latter step is mediated by integral membrane glycoproteins belonging to the superfamily of ATP-Binding Cassette (ABC) transporters. A subfamily, referred as ABCC, includes the famous/infamous cystic fibrosis transmembrane regulator (CFTR), the sulfonylurea receptors (SUR 1 and 2), and the multidrug resistance-associated proteins (MRPs). The name of the MRPs refers to their potential role in clinical multidrug resistance, a phenomenon that hinders the effective chemotherapy of tumors. The MRPs that have been functionally characterized so far share the property of ATP-dependent export pumps for conjugates with glutathione (GSH), glucuronate or sulfate. MRP1 and MRP2 are also mediating the cotransport of unconjugated amphiphilic compounds together with free GSH. MRP3 preferentially transports glucuronides but not glutathione S-conjugates or free GSH. MRP1 and MRP2 also contribute to the control of the intracellular glutathione disulfide (GSSG) level. Although these proteins are low affinity GSSG transporters, they can play essential role in response to oxidative stress when the activity of GSSG reductase becomes rate limiting. The human MRP4, MRP5 and MRP6 have only partially been characterized. However, it has been revealed that MRP4 can function as an efflux pump for cyclic nucleotides and nucleoside analogues, used as anti-HIV drugs. MRP5 also transports GSH conjugates, nucleoside analogues, and possibly heavy metal complexes. Transport of glutathione S-conjugates mediated by MRP6, the mutation of which causes pseudoxantoma elasticum, has recently been shown. In summary, numerous members of the multidrug resistance-associated protein family serve as export pumps that prevent the accumulation of anionic conjugates and GSSG in the cytoplasm, and play, therefore, an essential role in detoxification and defense against oxidative stress. PMID- 12897434 TI - Detoxification of electrophilic compounds by glutathione S-transferase catalysis: determinants of individual response to chemical carcinogens and chemotherapeutic drugs? AB - The glutathione S-transferases (GSTs) catalyze the GSH-dependent detoxification of reactive electrophiles such as genotoxic chemical carcinogens and cytotoxic chemotherapeutic agents. Allelic polymorphism in the GSTs has been used to investigate the hypothesis that GSTs are involved in susceptibility to human cancers. Such studies have resulted in low penetrance, high prevalence associations between cancer risk and GST polymorphisms. By examination of interindividual variation of GST expression it becomes clear that GST genotype alone is not an accurate predictor of GST expression. GST expression is tissue specific and interindividual variation of expression is at least 7-fold in normal tissues. Thus, populations of the same genotype are actually heterogeneous as regards expression. Similarly, polymorphisms are not effective in all tissues and GST induction is not independent of genotype. Mechanistic models for chemical aspects of colorectal cancer and chemotherapy for breast cancer demonstrate some of the ways by which such interactions can be studied and the potential for future studies. PMID- 12897435 TI - Transcriptional regulation of glutathione S-transferase P1-1 in human leukemia. PMID- 12897436 TI - The role of gamma-glutamyl transpeptidase in the biosynthesis of glutathione. PMID- 12897437 TI - Role of gamma-glutamyltransferase in the homeostasis of glutathione during oxidative and nitrosative stress. PMID- 12897438 TI - The importance of gamma-glutamyl transferase in lung glutathione homeostasis and antioxidant defense. PMID- 12897439 TI - The role of gamma-glutamyltranspeptidase in the metabolism and cytotoxicity of 4 hydroxynonenal-glutathione conjugate: evidence and hypothesis. PMID- 12897440 TI - Gamma-glutamyltransferase-dependent prooxidant reactions: a factor in multiple processes. PMID- 12897441 TI - Serum gamma-glutamyl transpeptidase: a prognostic marker in cardiovascular diseases. PMID- 12897442 TI - Lipoic acid: a multifunctional antioxidant. PMID- 12897443 TI - Is glutathione an important neuroprotective effector molecule against amyloid beta toxicity? PMID- 12897444 TI - Antioxidants in cancer therapy: is there a rationale to recommend antioxidants during cancer therapy? PMID- 12897445 TI - Disulfide exchange in CD4. PMID- 12897446 TI - Molecular chaperones, stress proteins and redox homeostasis. AB - Protection against oxidative stress is highly interrelated with the function of the most ancient cellular defense system, the network of molecular chaperones, heat shock, or stress-proteins. These ubiquitous, conserved proteins help other proteins and macromolecules to fold or re-fold and reach their final, native conformation. Redox regulation of protein folding becomes especially important during the preparation of extracellular proteins to the outside oxidative milieu, which should take place in a gradual and step-by-step controlled manner in the endoplasmic reticulum or in the periplasm. Several chaperones, such as members of the Hsp33 family in yeast and the plethora of small heat shock proteins as well as one of the major chaperones, Hsp70 are able to act against cytoplasmic oxidative damage. Abrupt changes of cellular redox status lead to chaperone induction. The function of several chaperones is tightly regulated by the surrounding redox conditions. Moreover, our recent data suggest that chaperones may act as a central switchboard for the transmission of redox changes in the life of the cell. PMID- 12897447 TI - Reduction of the endoplasmic reticulum accompanies the oxidative damage of diabetes mellitus. AB - The endoplasmic reticulum (ER), similary to other subcompartments of the eukaryotic cell possesses a relatively oxidizing environment. The special milieu of ER lumen is important for many ER-specific processes (redox protein folding, glycoprotein synthesis, quality control of secreted proteins, antigen presentation, etc.). Despite of the vital importance of redox regulation in the ER, we have a surprisingly fragmented knowledge about the mechanisms responsible for the ER redox balance. Moreover, new observations on disulfide bridge synthesis and on glutathione functions urge us to revise our recent theories based on many indirect and in vitro results. We have also very little information about the effects of different pathological conditions on the thiol metabolism and redox folding in the ER. Examining the role of molecular chaperones in the cellular pathology of diabetes mellitus we found that the ER redox environment shifted to a more reducing state, which was followed by changes of the thiol metabolism and structural-functional changes of the protein machinery involved in the redox folding process in diabetes. The possible consequences of these unexpected changes are also discussed. PMID- 12897448 TI - Analytical developments in the assay of intra- and extracellular GSH homeostasis: specific protein S-glutathionylation, cellular GSH and mixed disulphide compartmentalisation and interstitial GSH redox balance. PMID- 12897449 TI - Signalling potential and protein modifying ability of physiological thiols. PMID- 12897450 TI - Redox signaling and the MAP kinase pathways. AB - The mitogen-activated protein (MAP) kinases are a large family of proline directed, serine/threonine kinases that require tyrosine and threonine phosphorylation of a TxY motif in the activation loop for activation through a phosphorylation cascade involving a MAPKKK, MAPKK and MAPK, often referred to as the MAP kinase module. Three separate such modules have been identified, based on the TxY motif of the MAP kinase and the dual-specificity kinases that strictly phosphorylate their specific TxY sequence. They are the extracellular signal regulated kinases (ERKs), c-jun N-terminal kinases (JNKs) and p38 MAPKs. The ERKs are mainly associated with proliferation and differentiation while the JNKs and p38MAP kinases regulate responses to cellular stresses. Redox homeostasis is critical for proper cellular function. While reactive oxygen species (ROS) and oxidative stress have been implicated in injury, a rapidly growing literature suggests that a transient increase in ROS levels is an important mediator of proliferation and results in activation of various signaling molecules and pathways, among which the MAP kinases. This review will summarize the role of ROS in MAP kinase activation in various systems, including in macrophages, cells of myeloid origin that play an essential role in inflammation and express a multi component NADPH oxidase that catalyzes the receptor-regulated production of ROS. PMID- 12897451 TI - Redox regulation of mitochondrial permeability transition: effects of uncoupler, lipoic acid and its positively charged analog LA-plus and selenium. PMID- 12897452 TI - Redox state of glutathione and thioredoxin in differentiation and apoptosis. PMID- 12897453 TI - Redox-regulation of DNA repair. PMID- 12897454 TI - Application of allelochemicals to agriculture. AB - The study of allelopathy has a long history, and its application to agricultural production has long been anticipated. Recently, researchers have found allelopathic plants that are now used as cover crops, and allelochemicals which may lead to new herbicides. This paper reviews three studies introduced in this symposium, and discusses the possible application of allelopathy to agriculture. PMID- 12897455 TI - Allelopathy in the natural and agricultural ecosystems and isolation of potent allelochemicals from Velvet bean (Mucuna pruriens) and Hairy vetch (Vicia villosa). AB - We have studied on allelopathy of plants and developed methods to identify the effective substances in root exudates, leaf leacheate, and volatile chemicals emitted from plants. We found traditional cover plants that show allelopathic activity are useful for weed control. It could eliminate the use of synthetic chemicals for this purpose. Allelopathy is a natural power of plants to protect themselves by producing natural organic chemicals. Some endemic plants in Asia, already known by farmers in the region, as either cover crops used in intercropping, hedgerow, or agroforestry, were found to possess strong allelopathic abilities. Our group identified several allelochemicals from these plants. These allelopathic cover crops, mostly leguminous plants, provide protein rich food, and grow easily without artificial fertilizers, herbicides, insecticides and fungicides. In this regards, these allelopathic cover crops could save food shortage in rural area, and are useful for environmental conservation. Screenings of allelopathic plants by specific bioassays and field tests have been conducted. Hairy vetch (Vicia villosa) and Velvet bean (Mucuna pruriens) are two promising species for the practical application of allelopathy. An amino acid, L-DOPA, unusual in plants, plays an important role as allelochemical in Velvet bean (Mucuna pruriens). Hairy vetch is the most promising cover plant for the weed control in orchard, vegetable and rice production and even for landscape amendment in abandoned field in Japan. We have isolated "cyanamide", a well known nitrogen fertilizer, from Hairy vetch. This is the first finding of naturally produced cyanamide in the world. PMID- 12897456 TI - Reduced allelopathic inhibition of lettuce (Lactuca sativa) growth caused by velvet bean (Mucuna pruriens) under 3D-clinorotation. AB - Allelopathy between Mucuna pruriens (velvet bean) and Lactuca sativa (lettuce) was studied under 3D-clinorotation. Growth of both roots and shoots of lettuce seedlings was suppressed by the presence of velvet bean. The degree of suppression was less on the clinostat compared to the normal static earth gravity. L-DOPA (L-3, 4-dihydroxyphenylalanine) is known to be a major substance in allelopathy of velvet bean. Amount of L-DOPA diffused out from a sintered filter paper into agar medium was compared between clinorotation and control group, and found no significant difference. It was concluded that some factors related to release, transport, and sensing phenomena of allelopathic substances may be responsible to the new findings in this study. PMID- 12897457 TI - Allelopathy as a new strategy for sustainable ecosystems development. AB - Natural products involved in plant-plant and plant-microorganism ecological interaction (Allelochemicals) are an important potential source for alternative agrochemicals and pharmaceuticals, in order to solve the many problems derived from inadequate culture practices and abuse of synthetic herbicides. Isolation, structural determination, bioassay techniques and applicability for these compounds in crop protection and pharmaceutical research are discussed, and future trends on Allelochemicals applications are examined. The new strategies for sustainable ecosystems controlled by allelochemicals offer a particular interest for the development of human bases in space, since these products can stimulate or inhibit plant germination and growth, and permit to develop crops with low residue amounts in water, facilitating wastewater treatment and recycling. PMID- 12897458 TI - Flavonoid function and activity to plants and other organisms. AB - Flavonoid compounds distribute widely in vascular plants and Bryophytes, and ca. 5,000 kinds have been reported as naturally occurring substances. Many biological activities of the flavonoids were found until now. They include pollinator attractants, oviposition stimulants, feeding attractants and deterrents, allelopathy and phytoalexins. This paper reviews function and activity of flavonoids against plants and other organisms. PMID- 12897459 TI - Interaction between Japanese flowering cherry trees and some wild animals observed during physiological experiment in fields. AB - We have studied the weeping habit of Japanese flowering cherry tree in the field of Tama Forest Science Garden, Forestry and Forest Products Research Institute at the foot of Mt. Takao. Since cherry trees at various age were the materials for our plant physiology experiments, our studies were conducted in the fields where we experienced certain difficulties. Even under such difficult environment that was rather unexpected and uncontrollable, we could obtain fruitful results on the growth of cherry tree, and found them scientifically significant, especially in terms of biological effects of gravity on earth. Moreover, a lot of interesting interactions of cherry trees with various kinds of animals were observed in parallel to the plant physiology. PMID- 12897460 TI - An approach for the conquest of the sugi pollinosis using plant growth regulation. AB - The sugi (Cryptomeria japonica) pollinosis becomes representative allergic disease in early spring in Japan. However, effective treatment for the sugi pollinosis and countermeasure against pollen of C. japonica at its source have not been developed in a practical sense. In this paper, the research aiming to prevent dispersion of pollen of the C. japonica is introduced on application and practical application to the field from the laboratory using the growth regulation of the plant. We found that formation of male flower bud in C. japonica could be suppressed by TNE, since the 3 beta-hydroxylase is inhibited by the action of Trinexysapacethyl, TNE. PMID- 12897461 TI - Engineering of closed ecological system in space and inter-organismal interactions. AB - Space agriculture is a concept of synthesis and operation of closed ecological system for controlling living environment and supplying materials in order to sustain life and to meet metabolic needs of space crew. It revitalizes metabolites and other excretion of crew for their recycled usage. It is an advanced concept for life support for long and large scaled manned space missions, where open loop system for materials cycle is not feasible to apply. Several issues, such as cost-benefit analysis with considering benefits of amenity and psychological factors of crew, are discussed in this essay, together with reliability and productivity of biological systems. Studies on plant physiology for inter-organismal interaction contribute to design work for space agriculture, and associate with our engagements to our future, sustainable development of our civilization both on the earth and extending to outer space. PMID- 12897462 TI - Development of the anti-gravitational system in land plants and its implication for the interaction between plants and other organisms. AB - After they first went ashore during the Silurian epoch, plants have developed the anti-gravitational system to survive under terrestrial environment with the strong gravitational force. The cell wall acts as a principal component of the anti-gravitational system in plants, probably with the aid of links to the plasma membrane and the cytoskeleton. The cell wall has well developed in land plants and often represents more than 90% of the dry weight of the plant. The development of the cell wall has greatly influenced the interaction between plants and other organisms, such as feeding, sheltering, invasion, and symbiosis, and has been involved in the regulation of the global environment throughout the evolution. PMID- 12897463 TI - Growth of endophyte, Neotyphodium, and its host plant, tall fescue (Festuca arundinacea), under 3D-clinorotation. AB - Growth of a filamentous fungus endophyte, Neotyphodium, and its host plant, tall fescue, Festuca arundinacea, was examined during the seed germination process under pseudo-microgravity [correction of micrgravity] generated by three dimensional (3D-) clinorotation. The shoot growth of tall fescue infected with the endophyte was remarkably suppressed on a 3D-clinostat compared with that of the ground control. Without being infected, shoot growth of tall fescue was not strongly affected by the 3D-clinorotation. Many aggregated hyphae were observed in the plant seed incubated for 1-day on the 3D-clinostat [correction of clinost] than in those kept on the ground. These results indicate that the clinorotation induces responses in the endophyte and its host plant different from those under normal gravity. PMID- 12897464 TI - Comparative analysis of the angiotensin-II receptor in placental vascular endothelial cells in preeclamptic and normotensive patients. AB - The aim of this study was to compare the immunoreactivity of angiotensin-II receptor type 1 (AT1R) in the vascular endothelial cells of human placental vessels in normal and preeclamptic pregnancies. Immunohistochemistry was used to examine the localization of AT1R in the vascular endothelium of human placental vessels. In preeclamptic patients without intrauterine growth retardation (IUGR) the mean histological index H score was significantly higher, but significantly decreased in patients with preeclampsia complicated by IUGR in comparison with the control group. Immunolocalization of AT1R in vascular endothelial cells in our study supports the view that angiotensin II may play a role in the regulation of vascular tone and vascular resistance and the actions of other vasoactive factors, vasodilators or vasoconstrictors. However, our results confirmed that AT1R immunoreactivity is elevated in vascular endothelial cells of human placenta from pregnancies complicated by preeclampsia, suggesting a higher activity of the renin-angiotensin-aldosterone system (RAAS) in women with preeclampsia. On the other hand reduced immunoreactivity of AT1R in placental vascular endothelial cells in pregnancy complicated by preeclampsia with IUGR may result from chronic higher RAAS activity and may suggest the decreased ability to compensate and the inability to restore the normal balance between vasodilators and vasoconstrictors. These results may also reflect destructive changes and dysfunction of the vascular endothelium in preeclamptic pregnancy with IUGR. PMID- 12897465 TI - Effects of an angiotensin II receptor antagonist and angiotensin-converting enzyme inhibitors on burst forming units-erythroid in chronic hemodialysis patients. AB - BACKGROUND: Angiotensin-converting enzyme (ACE) inhibitors have been reported to reduce the response to erythropoietin (EPO) administration in chronic hemodialysis patients, but the mechanism for this effect has not yet been clarified. To clarify the mechanism of ACE inhibitors- and angiotensin II type 1 (AT1) receptor antagonist-induced anemia in hemodialysis patients, we examined the effect of ACE inhibitors and AT1 receptor antagonist on burst-forming units erythroid (BFU-E) in the peripheral blood of hemodialysis patients and healthy controls in vitro. METHODS: Peripheral blood mononuclear cells (PBMNCs) were isolated by gradient centrifugation from 10 patients on regular hemodialysis and 7 healthy control volunteers. A colony assay of hematopoietic progenitors was performed using the methylcellulose culture system. PBMNCs of 1 or 2 x 10(5) were plated in a medium containing EPO with various concentrations of ACE inhibitors or AT1 receptor antagonist and incubated for 14 days. Colonies of BFU-E were counted under an inverted microscope. RESULTS: The PBMNCs from the chronic hemodialysis patients formed fewer BFU-Es than those from healthy volunteers. AT1 receptor antagonist in both healthy volunteers and hemodialysis patients suppressed the number of BFU-Es. The ACE inhibitors produced a smaller effect than the AT1 receptor antagonist. CONCLUSION: AT1 receptor blockade can directly inhibit erythropoiesis in vitro. PMID- 12897466 TI - Emerging technology in spine: should we rethink the past or move forward in spite of the past? PMID- 12897467 TI - Interbody cage devices. AB - STUDY DESIGN: A literature review was conducted of basic science research and clinical experiences describing the use of interbody cage devices for the management of degenerative spinal abnormalities. OBJECTIVES: To summarize current knowledge regarding the use of interbody fusion cages. SUMMARY OF BACKGROUND DATA: Degenerative conditions of the lumbar and cervical spine are a major societal expense and a leading cause of disability. Fusion surgery may be used to treat patients with some of these conditions. During the past decade, interbody cages have been popularized as a useful fusion technique with high rates of clinical and radiographic success reported. Cages may be implanted using a variety of surgical approaches to the disc space and can be used alone or with supplemental posterior fixation. METHODS: A literature review of biomechanical, biologic, and clinical studies of threaded interbody cages was performed. RESULTS: Interbody cages have been shown to successfully promote fusion in a variety of animal models. In biomechanical studies, anteriorly placed threaded cages significantly stabilize the motion segment in all directions except extension. Posteriorly placed cages provide less stability as a result of the facetectomy required for placement of an appropriately sized device. Successful clinical and radiographic results have been reported with the use of interbody cages. Most reported cage failures are the result of technical difficulties with implantation or poor patient selection. Accurate radiographic assessment of fusion in the presence of a metal interbody cage remains challenging, and studies evaluating alternate biomaterial cages are underway. CONCLUSION: Interbody cages are a useful technique for achieving spinal fusion and have been shown to have an acceptable clinical success rate in appropriately selected patients. PMID- 12897468 TI - Intradiscal electrothermal therapy: a treatment option in patients with internal disc disruption. AB - STUDY DESIGN: A literature review was conducted. OBJECTIVES: To review the anatomy, pathophysiology, diagnosis, procedure, and clinical results of intradiscal electrothermal therapy (IDET). SUMMARY OF BACKGROUND DATA: Low back pain is a major physical and socioeconomic entity. A significant percentage of low back pain is attributable to internal disc disruption. The management of internal disc disruption has traditionally been limited to either conservative treatment or spinal fusion. IDET has been performed as an alternative to these therapies. METHODS: The available literature was reviewed. RESULTS: Scientific data regarding the pathophysiology, biologic effects, and clinical results are relatively scarce. Early biomechanical and histologic investigations into the effects of IDET are conflicting. However, in early prospective human trials, IDET seems to provide some benefit with little risk. CONCLUSIONS: IDET is potentially beneficial treatment for internal disc disruption in carefully selected patients as an alternative to spinal fusion. More basic science and clinical research with long-term follow-up evaluation is necessary. PMID- 12897469 TI - Intervertebral disc prostheses. AB - STUDY DESIGN: This article is based on a comprehensive review of the literature related to intervertebral disc prostheses. OBJECTIVE: To compile an overview of the results and complications related to various types of lumbar and cervical disc replacements. SUMMARY OF BACKGROUND DATA: A functional disc prosthesis has been sought since the 1950s. Although there were a few early attempts, disc replacement did not become a viable therapy until the late 1980s. Devices that were used in Europe at that time are now being evaluated in the United States in clinical trials. Cervical disc replacements are currently in the early stages of development and evaluation. METHODS: A thorough literature search was conducted to identify articles published on intervertebral disc prostheses. Proceedings from conferences were reviewed as well. Articles were classified by topics, including lumbar total disc replacement, disc nucleus replacement, and cervical disc replacement. RESULTS: Results of total disc replacement are favorable, with 63-85% of patients having a good outcome. As with any procedure, issues related to patient selection, operative technique, and device size selection were learned from the early experiences with these devices. There has been only one disc nucleus replacement used in an appreciable number of patients. This has yielded good results, but there have been problems related to device displacement. There are increasing reports on the use of cervical disc replacements, but they are still in the early evaluation phases. However, as with the lumbar counterparts, new designs are being evaluated. DISCUSSION: Although still in the early phases of development, functional disc replacement will become a part of the spine surgeon's armamentarium for the treatment of patients with disc-related pain unresponsive to nonoperative care. The future in this area is exciting and bright with the introduction of new materials and new designs for implants. However, as with any new technology, there must be careful consideration of the safety of the devices, and their effectiveness will only be determined in long-term follow-up studies. PMID- 12897470 TI - Summary statement: emerging techniques for treatment of degenerative lumbar disc disease. AB - Emerging techniques for management of degenerative disc disease include modern fusion interbody cage devices, intradiscal electrothermal therapy, and disc prostheses among others. The management of discogenic low back pain is controversial: its accurate diagnosis is difficult, and treatment is inconsistent. Before wide application, the advantages and disadvantages of each procedure must be considered in lieu of inadequate study design and the lack of sufficient long-term outcome studies. PMID- 12897471 TI - Minimally invasive lumbar fusion. AB - STUDY DESIGN: Review article. OBJECTIVES: To provide an overview of current techniques for minimally invasive lumbar fusion. SUMMARY OF BACKGROUND DATA: Minimally invasive techniques have revolutionized the management of pathologic conditions in various surgical disciplines. Although these same principles have been used in the treatment of lumbar disc disease for many years, minimally invasive lumbar fusion procedures have only recently been developed. The goals of these procedures are to reduce the approach-related morbidity associated with traditional lumbar fusion, yet allow the surgery to be performed in an effective and safe manner. METHODS: The authors' clinical experience with minimally invasive lumbar fusion was reviewed, and the pertinent literature was surveyed. RESULTS: Minimally invasive approaches have been developed for common lumbar procedures such as anterior and posterior interbody fusion, posterolateral onlay fusion, and internal fixation. As with all new surgical techniques, minimally invasive lumbar fusion has a learning curve. As well, there are benefits and disadvantages associated with each technique. However, because these techniques are new and evolving, evidence to support their potential benefits is largely anecdotal. Additionally, there are few long-term studies to document clinical outcomes. CONCLUSIONS: Preliminary clinical results suggest that minimally invasive lumbar fusion will have a beneficial impact on the care of patients with spinal disorders. Outcome studies with long-term follow-up will be necessary to validate its success and allow minimally invasive lumbar fusion to become more widely accepted. PMID- 12897472 TI - Anterior endoscopic discectomy and fusion for adolescent idiopathic scoliosis. AB - STUDY DESIGN: A review of adolescent patients with idiopathic scoliosis undergoing endoscopic release and spinal fusion. OBJECTIVE: To describe the indications, techniques, results, and complications of thoracic anterior endoscopic scoliosis surgery. SUMMARY OF BACKGROUND DATA: Anterior endoscopic treatment of thoracic adolescent idiopathic scoliosis has become an alternative method of surgical treatment. METHODS: Twenty-one patients with adolescent idiopathic scoliosis have undergone a thoracic anterior endoscopic release and fusion followed by posterior instrumentation and fusion. Indications for the endoscopic fusion were large curve magnitude, skeletal immaturity, and/or thoracic hyperkyphosis. Eleven patients have undergone anterior endoscopic instrumentation and fusion for thoracic scoliosis curves between 45 degrees and 70 degrees, using a single screw/single rod construct and autogenous rib bone graft. RESULTS: Results from the anterior endoscopic release and fusion procedures followed by a posterior instrumentation and fusion had an average preoperative curve of 82 degrees (range, 41 degrees -125 degrees ), with postoperative correction to 28 degrees (range, 5 degrees -60 degrees ) showing 70% correction. For patients undergoing an anterior endoscopic instrumentation and fusion, the average preoperative Cobb measurement of 53 degrees (range, 44 degrees -62 degrees ) was corrected to an average 26 degrees (range, 18 degrees 38 degrees ) for an average correction rate of 51%. One patient undergoing an anterior endoscopic release was converted to an open procedure for end plate bony bleeding without sequelae. One patient with an anterior endoscopic instrumentation and fusion had revision anterior surgery for a distal set screw dislodgment and subsequent posterior instrumentation and fusion for pseudarthrosis. CONCLUSIONS: The use of both anterior endoscopic release and fusion combined with either anterior instrumentation or separate posterior instrumentation and fusion continues to evolve. Surgeons treating patients with these techniques must understand that there are specific indications for them and many technique options available to optimize surgical results. PMID- 12897473 TI - Minimally invasive fusion: summary statement. AB - Ideally, minimally invasive surgery (MIS) allows less extensive manipulation of surrounding tissues than a conventional open procedure while accomplishing the same goals and objectives at the target structure. Long-term follow-up combined with appropriate outcome measures are necessary to prove the safety and effectiveness of MIS. For MIS procedures to be widely adopted, they must have an acceptable learning curve. Special skills are needed and are beyond those of traditional open surgery, By definition, as compared with conventional open surgeries, minimally invasive procedures typically involve smaller incisions and less extensive surgical manipulation of the tissues that surround the target structure. Ideally, once the target structure has been reached, the minimally invasive procedure should accomplish the same goals and objectives as its open surgical counterpart. Thus, although minimally invasive surgeries are aimed at reducing the morbidity associated with open surgical approaches, they should not hinder the surgeon's ability to perform a successful operation. In other words, minimal invasion should not equate to minimally effective. PMID- 12897474 TI - Minimally invasive treatments of osteoporotic vertebral compression fractures. AB - STUDY DESIGN: A literature review of experiences with vertebroplasty and kyphoplasty for treating symptomatic, osteoporotic vertebral compression fractures (VCFs). OBJECTIVES: To summarize the advantages and disadvantages of kyphoplasty and vertebroplasty. SUMMARY OF BACKGROUND DATA: Osteoporotic VCFs are a leading cause of disability and morbidity in the elderly. The consequences of osteoporotic VCFs (pain and often progressive vertebral collapse with resultant spinal kyphosis) adversely affect quality of life, physical function, mental health, and survival. Vertebroplasty and kyphoplasty are minimally invasive procedures for treating painful fractures. Vertebroplasty entails the percutaneous injection of bone cement into the fractured vertebra in attempts to stabilize the fracture and reduce pain. Kyphoplasty addresses pain and kyphotic deformity by the percutaneous expansion of an inflatable bone tamp to effect fracture reduction before cement deposition in a fractured vertebra. METHODS: A literature review of surgical techniques, indications, clinical results, and complications for vertebroplasty and kyphoplasty. RESULTS: Studies of vertebroplasty and kyphoplasty have reported excellent pain relief and improved function in most patients with osteoporotic VCFs. Vertebroplasty has the advantage of being relatively quick and inexpensive. Kyphoplasty, while associated with increased cost and surgical time, offers the potential to improve spinal alignment. In addition, by creating an intravertebral cavity, kyphoplasty reduces the risk of extravertebral bone filler extravasation. CONCLUSIONS: Vertebroplasty and kyphoplasty are currently used to treat osteoporotic VCFs with successful short-term results. Prospective, randomized studies comparing these procedures to one another and comparing their long-term outcomes to conventional medical management are required to define precise roles of these exciting treatments in the spine physician's armamentarium. PMID- 12897475 TI - Intraoperative spinal navigation. AB - STUDY DESIGN: Review article. OBJECTIVES: To provide a detailed overview of current methods for intraoperative spinal navigation using image-guided surgical technology. SUMMARY OF BACKGROUND DATA: The development of novel intraoperative navigational techniques has been an important advancement in the field of spine surgery. These techniques, commonly referred to as image-guided surgery (IGS), provide simultaneous, multiplanar views of spinal anatomy. They can be used for detailed preoperative planning and allow the spinal surgeon to track the position of surgical instruments in real time. IGS technology can increase the accuracy of spinal instrumentation procedures and improve patient safety. METHODS: The relevant medical literature was reviewed, as was the authors' clinical and laboratory experience with intraoperative spinal navigation. RESULTS: Image guided spinal instrumentation procedures in the cervical, thoracic, and lumbar spine have lower rates of screw misplacement than do those performed without image guidance. In a typical IGS spinal procedure, surgical instruments are tracked in the operating room, and their positions are superimposed onto preoperatively acquired computed tomography scans (CT-based image guidance) or intraoperatively acquired fluoroscopic images (virtual fluoroscopy). A new development, the combination of isocentric C-arm fluoroscopy with computer assisted image guidance, allows the C-arm to create intraoperative CT images that can be used for image-guided navigation without the need for a surgeon-dependent registration step. Each of these technologies has distinct advantages and limitations. CONCLUSIONS: Intraoperative spinal navigation has advanced rapidly in recent years, beneficially affecting a variety of surgical procedures. Future technological developments will widen its clinical application and minimize its shortcomings. PMID- 12897476 TI - Cement augmentation of osteoporotic compression fractures and intraoperative navigation: summary statement. AB - Emerging techniques and technologies in treating spinal disorders appear to be very promising. Vertebroplasty and kyphoplasty are new techniques that have been developed for treating osteoporotic vertebral compression fractures with early promising clinical results. More sophisticated and technically advanced methods of surgical navigation are also being developed as spinal surgery moves toward less invasive or minimally invasive procedures. Cost effectiveness, complication rates, and clinical efficacy need to be better defined to determine the exact role of these emerging techniques and technologies. PMID- 12897477 TI - Bone morphogenetic proteins and spinal surgery. AB - STUDY DESIGN: A review of the literature concerning the use of recombinant human bone morphogenetic proteins 2 (rhBMP-2) and 7 (rhBMP-7) in spinal fusion. PURPOSE: To summarize the pertinent preclinical experiments that enabled regulated human clinical trials of recombinant bone morphogenetic proteins for spinal fusion and to update clinicians on the results of those trials. BACKGROUND: More than three decades of research involving thousands of scientists and academicians throughout the world have led to the clinical use of recombinant bone morphogenetic proteins for the treatment of spinal disease. METHODS: The published and presented scientific literature and the author's personal experience were examined. RESULTS AND CONCLUSIONS: Recent clinical data support the assertion that recombinant bone morphogenetic proteins can be used as complete bone graft substitutes in spinal fusion surgery. In some circumstances, the efficacy of these factors for inducing successful fusion is superior to that of autogenous bone graft. RhBMP-2 is shown to be efficacious in several fusion applications, including intervertebral and lumbar posterolateral. Similar efficacy for rhBMP-7 has not yet been demonstrated, although relevant clinical studies are currently under way. The availability of these biologic agents will improve our ability to predictably treat spinal disease and may facilitate the further development of less invasive surgical innovations. PMID- 12897478 TI - Gene therapy applications for spine fusion. AB - OBJECTIVE: This article reviews the potential utilization of various growth factors to enhance spinal fusion and outlines the principles of gene therapy and its application to spinal fusion surgery. SUMMARY OF BACKGROUND DATA: Gene therapy offers an exciting new way to potentially deliver growth factors locally in a targeted fashion with physiologic doses. In its current definition, gene therapy is defined as the use of nucleic acid transfer, either RNA or DNA, to treat or prevent a disease. The scope of gene therapy has expanded beyond its initial application as a method of replacing genetic defects, and its potential to facilitate spinal fusions is currently being evaluated. CONCLUSIONS: Gene therapy strategies for spine fusion are appealing because the setting is uniquely suited for genetic manipulation. The intervention is locally applied. Only a short duration of transgene response by the cells is necessary to establish a spine fusion, and a variety of osteoinductive growth factors have been identified and are available for use. Attempts at spine fusion using gene therapy in the lower animals have been successful using both in vivo and ex vivo approaches. Before human clinical trials can be established, further testing is required in more challenging animal models of bone induction such as nonhuman primates. Should a successful clinical program of gene therapy for spine fusion be established, the use of autograft and its associated morbidities could be eliminated. In fact, gene therapy offers the potential for minimally invasive applications that could bypass the need for an open procedure altogether. It is likely that gene therapy will be a powerful therapeutic tool for the spine surgeon in the new millennium. PMID- 12897479 TI - BMPs and gene therapy for spinal fusion: summary statement. AB - The use of biologic technologies to engender spinal fusion is at various stages of scientific and clinical development. Recombinant human bone morphogenetic protein 2 is now available for clinical use in anterior lumbar intervertebral fusion and has been extensively studied in other spinal fusion applications. Recombinant bone morphogenetic protein 7 is under examination in regulated clinical trials. Gene therapy technologies are in the preclinical stage of development. In time, biologic interventions will continue to capture greater roles in the treatment of spinal disease. PMID- 12897480 TI - Biological repair of intervertebral disc. PMID- 12897481 TI - Gene therapy applications for intervertebral disc degeneration. AB - STUDY DESIGN: Review of current approaches to gene therapy as a potential treatment modality for regeneration of the degenerating intervertebral disc. OBJECTIVES: Describe recent advances in the field of gene therapy pertaining to treatment of the degenerating intervertebral disc. SUMMARY OF BACKGROUND DATA: The potential application of gene therapy to the treatment of degenerative disc disease has been an area of exciting and active research. Several in vitro studies have documented the capacity of gene therapy to favorably modify the biologic functions of the intervertebral disc cells with the delivery of the cDNA for various growth factors, whereas in vivo studies have reinforced this potential in animal models. Currently, investigators are exploring the therapeutic potential of gene delivery of the cDNA for several promising factors. METHODS: Literature review. RESULTS/CONCLUSIONS: The application of gene therapy to the treatment of the degenerating intervertebral disc holds much clinical promise and continues to expand as an area of active research. The therapeutic potential of multiple agents, as well as possible vectors, are currently being investigated with encouraging results. PMID- 12897482 TI - Biologic treatment for intervertebral disc degeneration: summary statement. AB - In past decades, the biomechanics of intervertebral disc degeneration have received much attention. More recent research efforts have focused on understanding the biology of the disc degenerative process. The ability to manipulate disc architecture and biology by direct application of growth factors or by gene therapy techniques may present an opportunity to halt or slow disc degeneration. Preliminary studies using these strategies have confirmed their ability to replenish essential disc matrix components that are known to decrease with disc degeneration. PMID- 12897483 TI - A critical guide to case series reports. AB - OBJECTIVE: Provide guidance to investigators and authors regarding appropriate conduct and reporting of case-series studies. SUMMARY OF BACKGROUND DATA: Evidence-based practice has provided a substantial contribution to advancing clinical science. Many study designs have been critically examined, and the quality of the research literature has improved. A common study design in musculoskeletal medicine is the case series: a description of the course of patients over time. Case series can provide valuable information as to: case definition, trend analyses regarding outcomes, and clues as to causation. Case series cannot be used to draw inferences regarding treatment effect. METHODS: Examination of previous work on identification of characteristics of high quality study designs such as cohort studies; extending this work to case series. RESULTS: We identified draft characteristics that good case series studies should address: clearly defined study question; well- described study population; well described intervention; use of validated outcome measures; appropriate statistical analyses; well-described results; discussion/conclusions supported by the data presented; funding sources acknowledged. CONCLUSIONS: We propose these measures to authors and journal editors as one mechanism to improve the quality of the case series study. PMID- 12897484 TI - Changes in spontaneous behavior in rats exposed to experimental disc herniation are blocked by selective TNF-alpha inhibition. AB - STUDY DESIGN: Study of pain behavior in animals by observation of changes in spontaneous behavior. OBJECTIVES: To assess if selective inhibition of tumor necrosis factor alpha may reduce changes in spontaneous behavior induced by experimental disc herniation in the rat as previously reported. SUMMARY OF BACKGROUND DATA: It is known that the proinflammatory cytokine tumor necrosis factor alpha may play a key role for the nucleus pulposus-induced nerve dysfunction seen in experimental set ups. However, it is not known if tumor necrosis factor alpha is also involved in pain production induced by the same procedure. METHODS: Thirty-two rats were used for the study. Twenty-two rats had an L4-L5 disc incision combined with a displacement of the L4 dorsal root ganglion. Twelve of these rats received an intraperitoneal injection of 0.125 mL of 10 mg/mL Remicade, and the remaining 10 were left untreated. Ten rats only had the L4-L5 disc exposed and formed the control group. The day before surgery and days 1, 3, 7, 14, and 21 after surgery, the rats were videotaped from below during a 20-minute period. The duration of four specific behaviors were determined and compared between the three experimental groups at each time point. RESULTS: Similar to a previous study, the nontreated showed increased signs of focal pain behavior (rotation of the head towards the operated leg and lifting of the operated leg) during the first 7 postoperative days. Treatment with the tumor necrosis factor alpha-inhibitor infliximab significantly reduced this behavior. At day 14, there were no differences between the groups, and at day 21, the nontreated group displayed reduced locomotion and increased immobility, similar to previous observations. Tumor necrosis factor alpha inhibition also seemed to reduce these behaviors. CONCLUSIONS: The data of the study clearly indicate a role for tumor necrosis factor alpha in the studied behavior changes after experimental disc herniation in the rat. Clinical trials must be performed in order to assess if there may be a clinical use for tumor necrosis factor alpha inhibition in the treatment of sciatica due to disc herniation. PMID- 12897486 TI - Effects of prostaglandin E1, melatonin, and oxytetracycline on lipid peroxidation, antioxidant defense system, paraoxonase (PON1) activities, and homocysteine levels in an animal model of spinal cord injury. AB - STUDY DESIGN: Investigation of the effects of prostaglandin E1, melatonin, and oxytetracycline on lipid peroxidation, antioxidant and paraoxonase activities, and homocysteine levels in an experimental model of spinal cord injury. OBJECTIVES: To determine the antioxidant efficacy of prostaglandin E1, melatonin, and oxytetracycline and whether paraoxonase and homocysteine can be used as monitoring parameters in the acute oxidative stress of spinal cord injury. SUMMARY OF BACKGROUND DATA: Melatonin has been found useful in spinal cord injury in previous studies. No study exists investigating the effects of melatonin, prostaglandin E1, and oxytetracycline as well as the response type of paraoxonase enzyme and homocysteine levels in the acute oxidative stress of spinal cord injury. METHODS: Sixty-three male albino Wistar rats were anesthetized with 400 mg/kg chloral hydrate and divided into 5 groups. The G1 (n = 7) control group provided the baseline levels. G2-G5 underwent T3-T6 total laminectomies and spinal cord injuries by clip compression at the T4-T5 levels. Medications were applied to G3-G5 right after clip compression. Hence, G2 constituted laminectomy + injury, G3 laminectomy + injury + prostaglandin E1; G4 laminectomy + injury + melatonin, and G5 laminectomy + injury + oxytetracycline groups. Animals were decapitated either the first or fourth hour after injury. Spinal cord tissue and blood malonyldialdehyde and plasma homocysteine levels, plasma glutathione peroxidase, superoxide dismutase, paraoxonase activities were assayed. The SPSS 9.0 program was used for statistical analysis and graphics. Intergroup comparisons were made by Bonferroni corrected Mann Whitney U test (P < 0.025) and intragroups comparisons by Wilcoxon Rank test (P < 0.03). RESULTS: In injury groups, plasma homocysteine levels decreased and paraoxonase activities increased as erythrocyte superoxide dismutase levels and plasma glutathione peroxidase activities decreased in parallel to increases of tissue and blood malonyldialdehyde levels. These alterations were relatively suppressed by prostaglandin E1, melatonin, and oxytetracycline administrations in varying degrees. Melatonin was the most powerful agent, particularly at the fourth hour. Oxytetracycline was also effective, both at the first and fourth hour. Prostaglandin E1 was effective in comparison to injury group, but not as much as melatonin and oxytetracycline. CONCLUSIONS: Melatonin and oxytetracycline are effective in preventing lipid peroxidation in spinal cord injury. Paraoxonase and homocysteine can be used in monitoring the antioxidant defense system as well as superoxide dismutase and plasma glutathione peroxidase, both in injury and medicated groups. PMID- 12897487 TI - In vivo evaluation of bone marrow stromal-derived osteoblasts-porous calcium phosphate ceramic composites as bone graft substitute for lumbar intervertebral spinal fusion. AB - STUDY DESIGN: Autogenous bone marrow stromal-derived osteoblasts-porous calcium phosphate ceramic composites were constructed in vitro under cell culture for 48 hours and implanted as a bone graft substitute for lumbar intervertebral spinal fusion in rabbits. OBJECTIVES: To evaluate the efficacy of autogenous bone marrow stromal-derived osteoblasts-porous calcium phosphate ceramic composites as an alternative to autogenous graft materials in a lumbar interbody spinal fusion model. SUMMARY OF BACKGROUND DATA: Bone marrow contains a population of rare progenitor cells capable of differentiating into bone, cartilage, muscle, tendon, and other connective tissues. These cells can be induced and differentiated into osteogenic osteoblasts with addition of osteogenic supplements. Combining bone marrow stromal-derived osteoblasts with porous ceramics gave rise to bone tissue in subcutaneous sites and repaired critical size segmental femoral defects. Little work has been done in the spine to assess fusion rates and associated biomechanical characteristics. METHODS: Five experimental groups were evaluated: sham operation (Group I); porous calcium phosphate ceramics alone (Group II); autogenous tricortical iliac crest (Group III); bone marrow stromal-derived osteoblasts-calcium phosphate ceramic composites (Group IV); bone marrow stromal derived osteoblasts-calcium phosphate ceramic composites with rhBMP-2 (Group V). All rabbits were killed 12 weeks after surgery, and the spinal fusion segments underwent the evaluation of gross inspection, manual palpation, radiography, computed tomography, nondestructive biomechanical testing, and histologic analysis. RESULTS: Successful spinal fusion was achieved by manual palpation in 100% (6/6) of animals in Group IV and Group V, 66.7% (4/6) in Group III, 50% (3/6) in Group II, and 0% (0/6) in Group I. Radiographic studies showed that minimal disc height loss was observed with ceramic blocks than with autograft. Biomechanical testingconfirmed that spines from Group IV and Group V were statistically significantly stiffer in flexion, extension, left and right bending, and left and right torsion than Group III and Group II. Histologic analysis demonstrated a qualitative increase of bone formation in fusion mass in Group IV and Group V versus all other groups. The size of fusion mass and the stiffness of fusion segments were greatest in Group V. CONCLUSION: The results indicate that bone marrow stromal-derived osteoblasts-calcium phosphate ceramic composites may provide an alternative to autogenous graft materials for lumbar interbody spinal fusion. Adding recombinant human bone morphogenetic protein-2 into the composites may reinforce the biomechanical stiffness for spinal fusion segments. Porous calcium phosphate ceramics alone were not suitable as a bone graft substitute for lumbar interbody spinal fusion. PMID- 12897488 TI - Prevention of postlaminectomy epidural fibrosis using bioelastic materials. AB - STUDY DESIGN: The use of elastic protein-based polymers for the prevention of epidural fibrosis following lumbar spine laminectomy was investigated in a rabbit model. OBJECTIVES: To determine the safety and efficacy of two bioelastic polymers in matrix and gel forms as interpositional materials in preventing postlaminectomy epidural fibrosis. SUMMARY OF BACKGROUND DATA: Postlaminectomy epidural fibrosis complicates revision spine surgery and is implicated in cases of "failed back syndrome." Materials employed as mechanical barriers to limit tethering of neural elements by the fibrosis tissue have met with little success. A recent family of protein-based polymers, previously reported to prevent postoperative scarring and adhesions, may hold promise in treating this condition. METHODS: Sixteen female New Zealand White rabbits underwent laminectomy at L4 and L6. Two polymer compositions, each in membrane and gel forms, were implanted at a randomly assigned level in four rabbits each, with the remaining level serving as an internal control. The animals were killed at 8 weeks, and qualitative and quantitative histology and gross pathologic examination were performed for both the control and the experimental sites to assess the polymers' efficacy in preventing dorsal epidural fibrosis. RESULTS: The use of the polymers caused no adverse effects. Compared to the control sites, both polymers in either gel or membrane form significantly reduced the formation of epidural fibrosis and its area of contact with the dura postlaminectomy. However, no significant difference in efficacy was detected between either the polymers or their respective forms in preventing epidural fibrosis. CONCLUSIONS: The selected compositions of biosynthetic, bioelastic polymers were safe and effective in the limiting the direct contact and consequent tethering of the underlying neural elements by the postlaminectomy epidural fibrosis in rabbits. PMID- 12897489 TI - Significance of fixation of the vertebral column for spinal cord injury experiments. AB - STUDY DESIGN: Thoracic spinal cord transections were performed in adult rats. The animals were divided into two groups, with or without internal fixation of the involved vertebral column. Histologic and immunohistochemical studies were performed to compare the effect of internal fixation of the vertebral column. OBJECTIVES: To find out the aspects and extent of beneficial effects of vertebral column fixation for spinal cord repair. SUMMARY OF BACKGROUND DATA: Vertebral column fixation is a routine procedure in clinical spinal cord surgery. Paradoxically, most, if not all, animal spinal cord experiments seem to have ignored the importance of vertebral column fixation. During trunk movements, the vertebral column flexes to different directions, accompanied by bending of the spinal cord. Following spinal cord lesions, with frequent bending of the cord there will be repeated bleeding, inflammation, and other pathologic processes at the lesion site. Thus, the healing process will be hampered. The severity of the damages that will be brought about by bending of the cord is, to a certain degree, unpredictable. There will be rather big individual variations in injury and repair among the same type of experiments, rendering quantification and conclusion difficult. METHODS: Adult Sprague-Dawley rats were used. The thoracic spinal cord was transected. Strong stainless steel wires were used for internal fixation of the vertebral column. The histology of the horizontal sections of the spinal cord segment, which included the lesion site, was examined at the 14th postoperative day. The volumes of the secondary degeneration and meningeal scar, the gap between the borders of the proximal and distal stumps of the transected spinal cord, the thickness of the meningeal scar, the astrocytic reaction, and the abundance of regenerating nerve fibers at the lesion site were compared between the vertebral column fixed and nonfixed groups. Whenever possible, the results were evaluated quantitatively. RESULTS: In all these aspects, the internally fixed group was consistently far better than the unfixed group. The quantitative analyses were as follows (fixed/unfixed): 1)volume of secondary degeneration: 1.07 +/- 0.20/1.81 +/- 0.43 mm3 (P < 0.01); 2) volume of meningeal scar: 2.38 +/- 0.55/4.34 +/- 1.40 mm3 (P < 0.05); 3) distance between cord stumps: 1.38 +/- 0.34/2.35 +/- 0.79 mm (P < 0.05); 4) the mean thinnest dimension of the meningeal scar: 0.90 +/- 0.43/1.98 +/- 0.85 mm (P < 0.05). CONCLUSION: Vertebral column fixation is a crucial procedure for spinal cord animal experiments. PMID- 12897490 TI - Boston brace correction in idiopathic scoliosis: a biomechanical study. AB - STUDY DESIGN: To analyze Boston brace biomechanics, pressure measurements and finite element simulations were done on 12 adolescent idiopathic scoliosis patients. OBJECTIVES: The aim was to analyze the Boston brace effectiveness using a finite element model and experimental measurements. SUMMARY OF BACKGROUND DATA: There are not very many biomechanical studies of Boston brace effectiveness, and its biomechanical action is not completely understood. METHODS: This study was performed on 12 girls with scoliosis treated with the Boston brace system. The experimental protocol was composed of the acquisition of two sets of multiplanar radiographs with and without brace followed by the pressure acquisition at the brace-torso interface. A personalized finite element modeling of the trunk was generated from the 3D reconstruction of the patient's geometry. The brace treatment was simulated by the application of equivalent forces calculated from the pressure measurements. RESULTS: Two Boston brace force patterns were defined from the pressure measurements. The first one consisted of high right thoracic forces of 31-113 N, lumbar forces less than 47 N, and included a left thoracic extension working as a counter pad. The second one consisted of low thoracic forces less than 20 N, lumbar forces up to 70 N, without left thoracic extension. The simulations showed that the passive forces only produced a coronal Cobb angle correction up to 9 degrees, whereas real correction was up to 16 degrees. CONCLUSION: High thoracic pads reduced more effectively both thoracic and lumbar scoliotic curves than lumbar pads only. The study suggests that mechanisms other than brace pads produce correction and contribute to the force equilibrium within the brace. PMID- 12897491 TI - Static and dynamic analysis of five anterior instrumentation systems for thoracolumbar scoliosis. AB - STUDY DESIGN: A nondestructive biomechanical investigation among five anterior spinal instrumentation systems for scoliosis. OBJECTIVES: The purpose of this study is to analyze the static and dynamic biomechanical stability of five different systems. SUMMARY OF BACKGROUND DATA: Although a variety of anterior spinal instrumentation systems for scoliosis are available, very few attempts have been made at comparative biomechanical studies. METHODS: Thirty calf spines were underwent static biomechanical tests, including flexion-extension, axial rotation, and lateral bending loading modes in the multisegmental spinal model. Five anterior instrumentation systems included: 1) Texas Scottish Rite Hospital system; 2) Bad Wildungen Metz; 3) anterior ISOLA; 4) Cotrel-Dubousset Hoph; and 5) Kaneda Anterior Scoliosis System. The initial and postfatigue stability after a cyclic loading test were analyzed by measuring the range of motion at instrumented segments compared to the intact within the same specimen (% to intact). RESULTS: Two-rod systems showed a significant decrease in range of motion compared to one-rod systems in flexion-extension (P < 0.001) and axial rotation (P < 0.05). In lateral bending, all systems demonstrated a significant decrease in range of motion of less than 40% to the intact (P < 0.001). After cyclical loading test, all systems increased in range of motion. In flexion extension, one-rod systems depicted a significant increase in range of motion, compared to two-rod systems (P < 0.05). CONCLUSIONS: In the initial stability analysis, two-rod systems are superior to one-rod systems. For one-rod systems, repeated physiologic loading may result in reduced stability in flexion extension. PMID- 12897492 TI - Magnetic resonance imaging of the cervical ligaments in the absence of trauma. AB - STUDY DESIGN: Prospective study of cervical spine magnetic resonance imaging. OBJECTIVES: To determine the frequency of nonvisualization of the cervical spinal ligaments in patients who have not suffered acute spinal trauma. SUMMARY OF BACKGROUND DATA: The cervical ligaments appear on magnetic resonance studies as a thin hypointense line, the "black stripe." Discontinuity of this "black stripe" is considered a reliable sign of ligament rupture following acute spinal trauma. METHODS: Twenty patients (13 female and 7 male, mean age 45.5 years, range 28-58 years) undergoing magnetic resonance imaging for cervical degenerative disc disease were examined using sagittal T1- and T2-weighted fast spin echo sequences. The status of the anterior longitudinal ligaments and posterior longitudinal ligaments was assessed on T1-weighted images from C2 to T1 (total of 120 levels), the ligamentum flavum from C1 to T1 (140 levels), and the apical ligament on both T1 and T2 weighted images (20 levels). The presence and sites of degenerative change were also noted. RESULTS: On the T1-weighted images, only 74 79% of anterior longitudinal ligaments, 36-74% of posterior longitudinal ligaments, 63-65% of ligamentum flavums, and 35-60% of apical ligaments were visualized. On T2-weighted images, all the apical ligaments were seen. An association was identified between nonvisualization of the anterior longitudinal ligaments on T1 and the presence of anterior osteophytes. CONCLUSION: Discontinuity of the "black stripe" on a T1-weighted sequence cannot be used as a reliable isolated sign of ligament rupture in the setting of acute spinal trauma, because the spinal ligaments are commonly not visualized. PMID- 12897494 TI - Influence of cage geometry on sagittal alignment in instrumented posterior lumbar interbody fusion. AB - STUDY DESIGN: Retrospective radiographic evaluation of the sagittal alignment of the lumbar spine in patients undergoing short-segment instrumented posterior lumbar interbody fusion with cage systems of different shape. OBJECTIVES: To determine whether rectangular and wedge-shaped cages have a different influence on the sagittal alignment of the lumbar spine in patients undergoing short segment instrumented posterior lumbar interbody fusion. SUMMARY OF BACKGROUND DATA: Previous studies of sagittal alignment after posterior lumbar interbody fusion have focused on the impact of intraoperative patient, hip, and knee positioning, as well as instrumentation characteristics on sagittal posture. The influence of the cage shape on indexes of total and segmental sagittal alignment of the lumbar spine is yet unknown. METHODS: Forty-two patients having undergone instrumented short-segment posterior lumbar interbody fusion were reviewed retrospectively. Twenty-two patients (12 women and 10 men, 38-78 years of age) had posterior lumbar interbody fusion with rectangular cages. The fused segments were: 4 at L3-L4, 16 at L4-L5, 11 at L5-S1. Thirteen patients had single- and nine patients double-level fusion. Twenty patients (8 women and 12 men, 34-81 years of age) had posterior lumbar interbody fusion with wedge-shaped cages. The fused segments were: 4 at L3-L4, 15 at L4-L5, 11 at L5-S1. Ten patients had single- and 10 patients double-level fusion. Cages were packed with cancellous bone from the posterior iliac crest and/or bone fragments harvested by laminectomy. All patients had additional pedicle screw fixation. Pre- and postoperative standing lateral radiographs were assessed for segmental and lumbar lordosis as well lumbar and sacral tilt. Data were analyzed with repeated measures analysis of variance. RESULTS The mean follow-up period was 18 months with a minimum follow-up period of 14 months. Mean segmental lordosis of the fused segments showed significant changes between the two implant groups (P < 0.05). Segmental lordosis decreased in the rectangular cage group from 10 degrees to 2 degrees at L3-L4, from 10 degrees to 5 degrees at L4-L5, and from 9 degrees before to 6 degrees after fusion surgery at L5-S1. In the wedge-shaped cage group, segmental lordosis increased from 4 degrees to 7 degrees at L3-L4, from 2 degrees to 8 degrees at L4-L5, and from 9 degrees to 17 degrees at L5-S1. Analysis of changes in lumbar lordosis and lumbar and sacral tilt did not show significant differences though opposite trends: lumbar lordosis decreased from 55 degrees to 48 degrees in the rectangular cage group and increased from 45 degrees to 53 degrees in the wedge-shaped cage group. Lumbar tilt measured 98 degrees before and 102 degrees after surgery in the rectangular cage group and 97 degrees before and 94 degrees after surgery. Sacral tilt measured 44 degrees before and 40 degrees after surgery in the rectangular cage group and measured 42 degrees before and 45 degrees after surgery in the wedge-shaped cage group. CONCLUSIONS: The cage geometry has a significant impact on the alignment of the lumbar spine after instrumented posterior lumbar interbody fusion. With rectangular cages, lumbar lordosis and segmental lordosis of the segments fused decrease; sagittal balance is maintained by compensatory changes of the sacral tilt. Wedge-shaped cages significantly increase segmental lordosis, enhance lumbar lordosis, and therefore should be preferred for restoring sagittal alignment in instrumented posterior lumbar interbody fusion procedures. PMID- 12897495 TI - Temperature and thermal dose distributions during intradiscal electrothermal therapy in the cadaveric lumbar spine. AB - STUDY DESIGN: Human cadaveric lumbar spines were used to assess the temperature and thermal dose distribution during intradiscal electrothermal therapy in vitro. OBJECTIVES: To determine whether intradiscal electrothermal therapy produces adequate tissue temperatures to denature annular collagen or ablate nerve cells. SUMMARY OF BACKGROUND DATA: Several hypothesized mechanisms for the effect of intradiscal electrothermal therapy have been suggested and include: 1) shrinkage of the nucleus and/or the annulus fibrosus by contraction of collagen fibers; and 2) thermal ablation of sensitive nerve fibers in the outer annulus. METHODS: Intradiscal electrothermal therapy was performed using the standard clinical protocol on 12 lumbar specimens in a 37.0 degrees C water bath using the SpineCath by Oratec. Temperatures were recorded simultaneously at 40 different locations in the disc. Thermal dose (Equivalent Minutes 43.0 degrees C) was calculated at each temperature point. RESULTS: The highest temperature measured (out of 520 points) was 64.0 degrees C and was within 1 mm of the heating coil. Temperatures in excess of 60 degrees C were all within 1 to 2 mm of the intradiscal electrothermal therapy catheter surface, the 50 to 60 degrees C range extended approximately 6 mm, above 48 degrees C extended approximately 7 mm, and above 45 degrees C extended to approximately 10 mm. Less than 2% of points achieved temperatures sufficient for collagen denaturation (>60 degrees C). On average, 42.5% of points accumulated >250 Equivalent Minutes 43.0 degrees C, a conservative common dose threshold for thermal necrosis of cells. The time history of thermal measurements demonstrated that the disc temperature had not reached steady state by the end of the heating protocol (16.5 minutes). CONCLUSIONS: Except for a very limited margin (1-2 mm) around the catheter, the temperature necessary to induce collagen shrinkage was not observed within the disc. Temperatures sufficient to ablate nerves were developed in some areas but were not reliably produced in clinically relevant regions, such as the posterior annulus. These results suggest that beneficial clinical outcomes may be critically dependent on probe placement or other factors unknown. PMID- 12897497 TI - Management of fixed sagittal plane deformity: outcome of combined anterior and posterior surgery. AB - STUDY DESIGN: Retrospective study of consecutive patient series. OBJECTIVES: To review the radiographic and clinical results of patients with preoperative fixed sagittal imbalance treated with combined anterior and posterior arthrodesis, and to determine factors that predict clinical outcome. SUMMARY OF BACKGROUND DATA: Combined anterior and posterior arthrodesis of the spine is useful in the management of fixed deformity involving the coronal and sagittal planes. The specific indications for combined surgery in the patient with regional and global imbalance have not been well defined. METHODS: Retrospective review of 25 consecutive patients treated with combined anterior and posterior spinal arthrodesis. Inclusion criteria included a preoperative global sagittal imbalance of at least 5 cm. Outcome variables included radiographic measures of preoperative, postoperative, and follow-up films, and a clinical assessment using the Modified SRS Outcomes Instrument and a review of postoperative complications. RESULTS: Twenty-five consecutive cases were reviewed. Mean age was 58 years (range 38-77), and mean follow-up was 55 months (range 24-81) for clinical and 44.5 (range 24-81) months for radiographic outcome variables. The mean preoperative sagittal imbalance was 10.5 cm (range 5.2-23.3), which improved to 2.9 cm (range 0-12.6) after surgery, and was maintained as 3.3 cm (range 0-13.5) at follow-up. Mean lumbar lordosis was -23 degrees (range +40 to -47) before surgery, and increased to -42 degrees at follow-up (range -20 degrees to -60 degrees ), an increase of 19 degrees. Patients with preoperative regional hypolordosis in the lumbar spine that was corrected surgically had the highest postoperative scores. The mean score for patient satisfaction with surgical management was 4.45 (range 2.5-5). Correlation analysis of clinical outcome domains demonstrated that patient satisfaction correlated poorly with domains of pain (r = 0.37, P = 0.1) and function (r = 0.4, P = 0.09). Within the domains, self-image showed highest correlation with patient satisfaction (r = 0.65, P = 0.006) and total score (r = 0.89, P = 0.0001). CONCLUSIONS: Patients with global sagittal imbalance of the spine were effectively treated with a combined anterior and posterior arthrodesis as measured by radiographic parameters. Patient satisfaction with surgery, and overall clinical outcomes were best in cases that resulted in an increase in lumbar lordosis. The subset of patients with preoperative regional hypolordosis of the lumbar spine has better outcomes than those with preoperative lumbar lordosis in the physiologic range. PMID- 12897499 TI - The passive straight leg raising test in the diagnosis and treatment of lumbar disc herniation: a survey of United kingdom osteopathic opinion and clinical practice. AB - STUDY DESIGN: Postal questionnaire survey. OBJECTIVES: To carry out a confidential postal survey of United Kingdom osteopaths in order to record and assess their use of the passive straight leg raising test in the diagnosis of, and choice of, manipulation for lumbar disc herniation. The study also sought to determine whether an association existed between osteopaths' manipulation of suspected lumbar disc herniation and their use of the straight leg raising test, the length of their working hours, and their use of manipulation for the treatment of other lumbar conditions. SUMMARY OF BACKGROUND DATA: The literature is not agreed on important aspects of the straight leg raising test, or on the use of spinal manipulation for suspected lumbar disc herniation. This is thought to be the first study to investigate opinion and practice in a large group of spinal manipulators, in this case United Kingdom osteopaths. METHODS: A questionnaire was sent to all 1030 United Kingdom osteopaths registered with the General Osteopathic Council in January 2000. It comprised four sections: personal characteristics, professional characteristics, background to low back pain cases, details of straight leg raising test understanding and use within the diagnosis and treatment of lumbar disc herniation. RESULTS: A response rate of 44% was achieved. United Kingdom osteopaths' opinions of low back pain and lumbar disc herniation clinical presentations, details of straight leg raising test mode of action, procedure, and interpretation were in keeping with the literature. Fifty four percent of respondents sometimes employed manipulation in the treatment of lumbar disc herniation, but most of the others described the practice as "dangerous." The literature is similarly divided on the practice. Chi-square and Cramer V analysis implied that respondents were not influenced in choosing manipulation for lumbar disc herniation by their use of the straight leg raising test (chi2 = 4.002, df = 3, Cramer V = 0.0959, P = 0.261, alpha 0.05, n = 435). A moderate association implied that the frequency of use of such manipulation for all lumbar conditions influenced the choice of that treatment for lumbar disc herniation (chi2 = 81.808, df = 4, Cramer V = 0.4302, P < 0.001, alpha = 0.05, n = 442). There was also a weak association suggesting that hours worked per week influenced the choice of manipulation for lumbar disc herniation (chi2 = 9.840, df = 3, Cramer V = 0.1499, P = 0.020, alpha = 0.05, n = 438). CONCLUSIONS: Respondents to this survey frequently treated low back pain and often employed the straight leg raising test in its diagnosis. Their recognition of the clinical presentation of lumbar disc herniation and their use and understanding of the straight leg raising test were in keeping with the literature. Respondents were divided nearly equally between those who would expect patient benefit from the use of manipulation for lumbar disc herniation and those who criticized the practice. There is a need for further research into the clinical reasoning employed for the manipulative treatment of lumbar disc herniation. PMID- 12897500 TI - Trunk sway measures of postural stability during clinical balance tests in patients with chronic whiplash injury symptoms. AB - STUDY DESIGN: Trunk sway occurring during clinical stance and gait tasks was compared between a group of subjects with a chronic whiplash injury, resulting from an automobile collision, and a normal collective. OBJECTIVES: To examine if population specific trunk sway patterns for stance and gait could be identified for chronic whiplash injury patients. SUMMARY OF BACKGROUND DATA: Our previous work has established that it is possible to identify specific patterns of stance and gait deficits for vestibular loss (both acute and compensated) patients and those with Parkinson's disease. Our question was whether it was possible to use the same stance and gait tasks to identify patterns of trunk sway differences with respect to those of healthy subjects and individuals with a chronic whiplash injury. METHODS: Twenty-five subjects with history of whiplash injury and 170 healthy age-matched control subjects participated in the study. Trunk sway angular displacements in chronic whiplash patients were assessed for a number of stance and gait tasks similar to those of the Tinetti and Clinical Test of Sensory Interaction and Balance (CTSIB) protocols. We used a lightweight, easy-to attach, body-worn apparatus to measure trunk angular displacements and velocities in the roll (lateral) and the pitch (forward-backward) planes. RESULTS: Data analysis revealed several significant differences between the two groups. A pattern could be identified, showing greater trunk sway for stance tasks and for complex gait tasks that required task-specific gaze control such as walking up and down stairs. Trunk sway was less, however, for simple gait tasks that demanded large head movements but no task-specific gaze control, such as walking while rotating the head. CONCLUSIONS: Subjects who have a chronic whiplash injury show a characteristic pattern of trunk sway that is different from that of other patient groups with balance disorders. Balance was most unstable during gait involving task-specific head movements which possibly enhance a pathologic vestibulo-cervical interaction. PMID- 12897501 TI - The use of intradiscal antibiotics for discography: an in vitro study of gentamicin, cefazolin, and clindamycin. AB - STUDY DESIGN: In vitro determination of minimum inhibitory concentrations (MICs) of gentamicin, cefazolin, and clindamycin, alone and in combination with iohexol against laboratory strains of Eschericia coli B, Staphylococcus aureus, and Staphylococcus epidermidis. OBJECTIVE: To study the effects of iohexol on the efficacy of gentamicin, cefazolin, and clindamycin. SUMMARY OF BACKGROUND DATA: Prophylactic antibiotics have been advocated to prevent discitis following discography. Intravenous cefazolin administered before discography has been shown to penetrate the intervertebral disc. However, the use of systemic antibiotics for prophylaxis may lead to bacterial resistance. Intradiscal antibiotic administration is an attractive alternative to systemic antibiotic prophylaxis before discography, but there is no data documenting the efficacy of commonly used antibiotics in the presence of iohexol. METHODS: MICs were determined by adding standard concentrations of bacteria to serial dilutions of antibiotic with and without the addition of iohexol in Todd-Hewitt Broth medium. MICs were determined as the lowest concentration well that demonstrated inhibition of cell growth. RESULTS: Gentamicin, cefazolin, and clindamycin remain efficacious in the presence of iohexol. MICs were lower for cefazolin and gentamycin than for clindamycin. Iohexol alone also demonstrated some inhibition of cell growth. CONCLUSION: This study supports the use of intradiscal antibiotics for prophylaxis of disc space infection during discography. lntradiscal placement of antibiotic should obviate the need for systemic antibiotic prophylaxis and its attendant risk of generating antimicrobial resistance. PMID- 12897502 TI - Reliability, validity, and responsiveness of the short form 12-item survey (SF 12) in patients with back pain. AB - STUDY DESIGN: Secondary analysis of data collected from spine patients' normal clinic visits from 1998 to 2001. OBJECTIVE: To evaluate the reliability, validity, and responsiveness of the short form 12-item survey in patients with back pain. SUMMARY OF BACKGROUND DATA: The reliability, validity, and responsiveness of the short form 12-item survey in patients with back pain has not been previously evaluated. METHODS: Patients were asked to complete a comprehensive computerized survey questionnaire during their regular clinic visits. A total of 2520 patients who indicated in their first surveys that they had back pain were included in the study of the reliability and validity of the short form 12-item survey. Of these, 506 patients completed another survey within 3-6 months of follow-up and were included in the responsiveness evaluation. RESULTS: The two summary scales of the short form 12-item survey, physical component summary and mental component summary, demonstrated internal consistency reliability, with Cronbach alpha for both scales exceeding the recommended level of 0.70. Correlation of physical component summary and mental component summary with six other measures theoretically related or unrelated to these scales performed as expected without exception, demonstrating the construct validity of the short form 12-item survey. The responsiveness of the short form 12-item survey was supported by several pieces of evidence. First, the changes in physical component summary and mental component summary scores were correlated with the changes in back pain intensity. Second, for patients whose back pain improved, there was a significant increase in the follow-up physical component summary and mental component summary scores as compared to the baseline. Third, small to moderate effect size was observed for patients whose back pain became improved or became worse. CONCLUSIONS: The short form 12-item survey demonstrated good internal consistency reliability, construct validity, and responsiveness in patients with back pain. PMID- 12897503 TI - Color Doppler ultrasonography for evaluation of internal mammary artery application in adolescent female patients with right-convex thoracic idiopathic scoliosis. AB - STUDY DESIGN: Prospective comparative study. OBJECTIVE: To establish the use of Color Doppler Ultrasonography to investigate internal mammary artery. SUMMARY OF BACKGROUND DATA: Breast asymmetry in female adolescents with right convex idiopathic scoliosis was supposed to be linked with anatomic and functional asymmetry of the internal mammary artery that is the main supplier to the mammary gland. However, no measurements of anatomic and hemodynamic parameters of internal mammary artery have been made to justify or to reject the hypothesis of asymmetric blood flow volume to the breasts and costosternal junction in female adolescent scoliotics. Color Doppler Ultrasonography is a well established noninvasive method to assess vessel anatomy and hemodynamics. METHODS: Twenty female adolescents with right thoracic scoliosis and 16 comparable female individuals without spine deformity were examined with Color Doppler Ultrasonography to measure at the origin of internal mammary artery lumen diameter, cross sectional area, time average mean flow and flow volume per minute and were compared each other. RESULTS: The reliability of Color Doppler Ultrasonography was high and the intraobserver variability low (ANOVA, P = 0.92 0.94). There was no statistically significant difference in the ultrasonographic parameters of the internal mammary artery between right and left side in each individual as well as between scoliotics and controls. CONCLUSIONS: Color Doppler Ultrasonography applied to assess anatomic or hemodynamic blood flow parameters at the origin of internal mammary artery was proven a highly reliable method. Color Doppler Ultrasonography disclosed no side-differences, while there were no differences between scoliotics and controls. Thus, it seems that this study cannot justify previous theories for development of right thoracic scoliosis in female adolescents. PMID- 12897504 TI - Post-traumatic findings of the spine after earlier vertebral fractures in young patients. PMID- 12897505 TI - Immunolocalization of phagocytic cells in normal and degenerated intervertebral discs. PMID- 12897506 TI - Chronic Chlamydia pneumoniae infection increases the risk of occlusion of lumbar segmental arteries of patients with sciatica: a 3-year follow-up study. AB - STUDY DESIGN: A magnetic resonance imaging follow-up study of lumbar arteries among patients with sciatica with chronic Chlamydia pneumoniae infection. OBJECTIVE: To determine whether chronic infection causes occlusion of lumbar arteries. SUMMARY OF BACKGROUND DATA: C. pneumoniae infection is associated with coronary heart disease, and the infectious pathogen has also been detected in abdominal aortic aneurysms and in atherosclerotic plaques. No studies are available on the effect of this infectious agent on lumbar arteries. METHODS: Chronic infection was defined as persistent high positive immunoglobulin G and/or immunoglobulin A antibodies and/or the presence of immune complexes. The lumbar arteries, evaluated with two-dimensional time-of-flight magnetic resonance angiography, were scored as normal, narrowed, or occluded. The differences in the segmental and whole lumbar spine (segments L1-L4) sum of arterial occlusion at baseline and at 3 years, and the incidence of new arterial stenosis were compared in patients with and without chronic infection using the Kolmogorov-Smirnov test. RESULTS: Patients with chronic infection were more likely to be persistent smokers (P = 0.006), male (P = 0.04), and more obese (P = 0.02) compared to patients with normal antibody levels. They had significantly higher degree of arterial stenosis at L4 segment at baseline and at 3 years (P = 0.001 and 0.002, respectively), in the whole lumbar spine at baseline and at 3 years (P < 0.001 for both), and at L1 and L3 segments at 3 years (P = 0.013 for both). The incidence of new arterial stenosis was similar in both patient groups. Patients with chronic infection also had significantly higher grade of endplate degeneration at L4-L5 (P = 0.008). CONCLUSIONS: The results of this study suggest that chronic C. pneumoniae infection may induce stenosis of lumbar arteries. PMID- 12897507 TI - Management of the cervical esophagus and hypofarinx perforations complicating anterior cervical spine surgery. AB - STUDY DESIGN: Five cases of esophageal or pharyngeal perforation diagnosed during or after anterior cervical spine surgery are presented. OBJECTIVE: To outline a protocol for the early diagnosis and treatment of iatrogenic pharyngoesophageal perforations. SUMMARY OF BACKGROUND DATA: Pharyngoesophageal perforations after anterior cervical spine surgery are uncommon or rarely reported complications. They may have serious functional consequences, including death, if they are not diagnosed promptly and treated effectively. These potentially fatal conditions require a surgical and medical therapy. METHODS: Clinical course, diagnostic tools and guidelines for the management of five patients presenting esophagopharingeal perforations are illustrated. RESULTS: These five cases resulted in definitive healing of the laceration without functional consequences. CONCLUSION: We believe that awareness of these complications and their causes, prompt recognition of the symptoms and immediate and multimodality therapies are essential tools to achieve successful results. PMID- 12897508 TI - Total sacrectomy and reconstruction for sacral tumors. AB - STUDY DESIGN: Report of three patients in whom the lumbosacral junctions were successfully restored by spinal instrumentations after total sacrectomies. OBJECTIVES: To describe the surgical technique of the reconstruction of the continuity between the pelvic ring and spinal column by using a transpedicular and iliac screw system. SUMMARY OF BACKGROUND DATA: Although there have been case reports about reconstruction methods after total sacrectomy, biomechanical, and technical problems still remain unresolved. METHODS: Total sacrectomy was carried out in three cases: two with chordomas and one with a recurrent giant cell tumor. In the first case, reconstruction was achieved with Zielke transpedicular screw and rod system and a sacral rod. The other two patients were reconstructed using a transpedicular and iliac screw system and a sacral rod for bilateral fixation of the iliac wings. In the third patient, the vertical rods were connected to transverse rod with rod connectors. RESULTS: No instrumentation failure was observed, and the continuity between the pelvic wing and spinal column was established with the instrumentation and bone grafting. Although one patient died of metastatic chordoma, the lumbosacral junction was successfully reconstructed with the instrumentation. The other two patients could stand with double crutches 13 and 2 years after surgery, respectively. CONCLUSIONS: Total sacrectomy is a feasible operation for primary malignant tumors involving the entire sacrum. Reconstruction of the union between the lumbar spine and the ilia with spinal instrumentation achieves stabilization suitable for ambulation. PMID- 12897509 TI - Two-stage (posterior and anterior) surgical treatment using posterior spinal instrumentation for pyogenic and tuberculotic spondylitis. AB - STUDY DESIGN: A retrospective analysis was performed of the clinical outcomes of patients with pyogenic or tuberculotic spondylitis who were treated with two stage surgery (first stage: placement of posterior instrumentation; second stage: anterior debridement and bone grafting). OBJECTIVE: To evaluate the clinical outcomes of the abovementioned two-stage surgical treatment for pyogenic or tuberculotic spondylitis. SUMMARY OF BACKGROUND DATA: Although several methods of surgical treatment for pyogenic and tuberculotic spondylitis have been reported, there have been few reports of two-stage surgical treatment. METHODS: Eight patients (7 male, 1 female) with pyogenic or tuberculotic spondylitis (pyogenic: 6; tuberculotic: 2) were treated by two-stage surgery (first: placement of posterior instrumentation, second: anterior debridement and bone graft). Age at the time of surgery was 63.5 +/- 9.91 years (average +/- SD) (range: 47 to 77 years). Most of the patients had systemic problems, such as pneumonia, diabetes mellitus, or chronic renal failure. First, posterior spinal instrumentation was placed. Then, anterior debridement and bone grafting were performed. Patients were evaluated before and after surgery in terms of pain level, hematologic parameters, neurologic status, and Barthel index. RESULTS: Average duration of surgery for both procedures was less than 4 hours. Changes in the pain level, blood parameters, and Barthel index demonstrated significant clinical improvement in all patients. Posterior wound infection occurred in two patients who were in poor general condition. CONCLUSIONS: This two-stage surgical treatment for pyogenic or tuberculotic spondylitis provided satisfactory results and can also be used in patients who are in poor general condition. PMID- 12897510 TI - An unusual case of CV junction tuberculosis presenting with quadriplegia. AB - STUDY DESIGN: Isolated tubercular involvement of craniovertebral junction in a human immunodeficiency virus-positive patient causing paraplegia and sudden death with radiologic features is presented. OBJECTIVES: Isolated involvement of craniovertebral junction by tuberculosis causing quadriparesis is a rare entity. The role of imaging features is presented in diagnosis of craniovertebral junction tuberculosis, which is a treatable disease. Early detection of this entity with prompt treatment can prevent a fatal outcome. SUMMARY OF BACKGROUND DATA: Tuberculosis of the cervical spine is a rare and potentially dangerous manifestation of extrapulmonary tuberculosis. The incidence is probably less than 1% of all cases of spinal tuberculosis. However, in the developing countries this constitutes an increasingly important cause of craniovertebral junction instability and cervicomedullary compression. Most of the patients present with pain in the neck and local tenderness. Neurologic deficits of varying degrees have been reported in 24-40% of cases of craniovertebral junction tuberculosis. Quadriplegia followed by sudden death is exceptional (as seen in our case). The incidence of craniovertebral junction tuberculosis in immunocompromised patients is not known. Dramatic recovery is possible if craniovertebral junction tuberculosis is detected early in its course. Prompt medical and surgical treatment may avert a potential catastrophic event in such cases. Imaging methods such as computed tomography and magnetic resonance imaging are diagnostic of this condition and aid in the detection and prompt treatment of the same. METHOD: Frontal radiograph of the cervical spine and chest, and lateral view of cervical spine followed by plain and contrast enhanced computed tomography scan of the cervical spine was performed to detect the lesion. RESULT: These radiographic features were correlated with the clinical findings. The computed tomography findings of bone destruction, prevertebral and extradural peripherally enhancing soft tissue and infiltrating opacities in the lung apexes were consistent with tuberculosis. CONCLUSIONS: The computed tomography findings described in this report are very specific for tuberculosis of the craniovertebral junction. Clinical and radiologic correlation could help in making the early diagnosis and prompt treatment possible. PMID- 12897511 TI - Sternal wound debridement and muscle flap reconstruction: functional implications. AB - The mortality rate for poststernotomy infection, which occurs in as many as 5% of median sternotomy incisions after cardiovascular surgery, was 37.5% until sternal debridement with muscle or omental flap reconstruction became the standard treatment for this postoperative complication and lowered the mortality rate to just more than 5%. There are few reports in the literature of physical functional deficits and long-term outcome resulting from such reconstruction. The authors evaluated two groups of patients who had undergone coronary bypass surgery at least 6 months earlier. One group had no postoperative complications; the other group had developed marked sternal wound infections that required debridement and pectoralis major or rectus abdominis muscle reconstruction. Both groups underwent pectoralis and rectus muscle strength testing, evaluation of pain and ability to perform those activities of daily living that are dependent on pectoral and rectus muscle function, and completed self-assessment questionnaires. Differences between the two groups were significant (p<0.05) with regard to pain and patient satisfaction with appearance and general functional capacity. Pectoral muscle function and strength were significantly different in patients in whom that muscle was transposed. Rectus muscle strength was not affected by the transposition of a single rectus muscle. Physical morbidity and loss of strength seemed to be related directly to loss of sternal stability stemming from marked infection and debridement rather than from loss of the muscles used in reconstruction. PMID- 12897512 TI - Response to letter by Dr. Yavuzer. PMID- 12897513 TI - Reconstruction after extirpation of sacral malignancies. AB - Defects after extirpation of either sacral or rectal tumors often present a reconstructive challenge to plastics surgeons. Because of their relative infrequency, management guidelines, in the authors' opinion, have been overlooked. They think that successful, comprehensive treatment lends itself to an integrated team approach. They review their experience with immediate reconstruction after total sacrectomy for sacral malignancies performed between 1996 and 2001. Medical records were reviewed retrospectively for the surgical procedure, postoperative complications, and eventual outcome. A total of 9 patients underwent sacrectomy with a gluteus maximus flap for reconstruction. Six patients had a simultaneous omental flap for complete obliteration of the surgical defect. The authors' experience suggests that this combination of techniques is a reliable approach for reconstruction of these extensive surgical defects. PMID- 12897514 TI - Regional differences in ultrasonic assessment of subcutaneous fat thickness in the abdomen: effects on the TRAM flap. AB - The authors describe the results of fat thickness patterning of the abdominal sites in 50 patients, all of whom required breast reconstruction with a transverse rectus abdominis musculocutaneous flap. The thickness of the abdominal fat was measured at 12 anatomic locations with an ultrasonic instrument. The highest value of the subcutaneous fat thickness was 29.0+/-10.0 mm at a site 2 cm below the umbilicus at the center of the rectus abdominis muscle. The lowest value of the subcutaneous fat thickness was 17.8+/-7.6 mm at a site 2 cm above the umbilicus on the anterior superior iliac spine. Average subcutaneous fat thickness over the abdomen of 50 patients was 24.0+/-9.4 mm. There were 13 patients (group 1) who had an abdominal fat thickness of more than 30 mm, 19 patients (group 2) with an abdominal fat thickness less than 30 mm and more than 20 mm, and 18 patients (group 3) with an abdominal fat thickness less than 20 mm. Complications occurred in 12 of 50 flaps (24%). Among groups 1, 2, and 3 there was no significant difference (p<0.01) in the overall flap complications (15.4: 36.8: 16.7). In summary, subcutaneous fat thickness showed the higher value at the center of the abdomen and the lower value at the lateral site. Abdominal fat thickness is not a risk factor for necrosis of pedicled transverse rectus abdominis musculocutaneous flaps in patients who are thin, average, or mildly obese. Preoperative examination of the abdominal subcutaneous fat thickness should provide useful information for detailed simulation of a reconstructive operation. PMID- 12897515 TI - Initial experience with personal digital assistant-based reflectance photoplethysmograph for free tissue transfer monitoring. AB - Improved microsurgical technique has resulted in a high percentage of successful free tissue transfers. When a tissue transfer fails in the head and neck, however, the results are orocutaneous fistulas, carotid artery exposure, and deformity that adds morbidity, expense, and may delay adjuvant therapy. Postoperative monitoring of tissue perfusion can detect early problems in free tissue transfer that may allow for early intervention and salvage. The authors have demonstrated that reflectance photoplethysmography can detect perfusion changes in free tissue transfer within 5 minutes of a pedicle "insult" intraoperatively. Normative data for viable flaps from various donor sites have been established. The authors now report their initial experience with a newly developed reflectance photoplethysmograph based on a hand-held computer for routine clinical use. Their results are compared with a conventional surveillance protocol that included observation, bleeding to pin prick, and bedside duplex scanning of the vascular pedicle. In a series of 30 free tissue transfers (29 patients), there was one ischemic event (skin paddle loss only), which was detected by the monitor. The monitor was able to predict correctly (one flap) survival of a free tissue transfer even when duplex ultrasonic data were indicative of an absence of perfusion. Personal digital assistant-based photoplethysmography appears to be a promising device for bedside diagnosis of free tissue transfer viability or ischemia. PMID- 12897516 TI - Tendon function after replantation: prognostic factors and strategies to enhance total active motion. AB - The purpose of this study was to determine the important prognostic factors for recovery of tendon function as measured by total active movement (TAM) in patients undergoing digital replantation. More important, the authors wanted to establish which factors may be manipulated to maximize motion. A retrospective review of 48 patients (103 digital rays) who underwent replantation was performed. Average TAM for all digits was 129 deg. Zone 1 and zone 5 injuries had better TAM than injuries in zones 2, 3, and 4, which had TAM values not significantly different from one another. Avulsion injuries fared significantly worse than other mechanisms of injury. TAM values were not affected by age, type of bone fixation, number of arteries repaired, or number of digits injured. Digits with both the profundus and the superficialis tendons repaired had significantly better TAM values relative to one-tendon fingers. Similarly, fingers treated with an "early" mobilization regime also exhibited better movement. Small numbers of injured digits in some groups may have limited our ability to detect significant differences. PMID- 12897517 TI - Free tissue transfer: an advanced strategy for postinfection soft-tissue defects in the upper extremity. AB - Surgical treatment of severe, necrotizing infections frequently leave compound defects that require complex reconstructive procedures. In the upper extremity, local flap coverage is limited because of the size of the lesions. Regarding the results of early microsurgical coverage of complex postinfectious defects of the lower extremity, the goal of this study was to evaluate the role of free tissue transfer in the treatment of severe infections in the upper extremity. Between 1994 and 1999, 24 patients with major defects as a result of severe necrotizing infections in the upper extremity underwent free tissue transfer. Parameters assessed included the success of infection control, flap survival rate, salvage of the extremity, and an outcome analysis by the Disability of Arm-Shoulder-Hand score and a visual analog scale. Patient age ranged from 17 to 75 years (average age, 50.8 years). Previous treatment of 11 patients in outlying hospitals included 4.2 operative procedures and a delay of admission to the authors' unit of 89 days. The average defect size after debridement was 10.0 x 14.4 cm. Twenty four free flaps including 16 muscle or musculocutaneous flaps, 4 chimeric flaps from the subscapular system, and 4 osteocutaneous flaps were performed for reconstruction. The overall flap survival was 95.8%. One temporalis fascia flap (TPF) was lost as a result of vascular thrombosis, and three flaps underwent successful revision of the anastomoses. Eight patients required further minor surgical treatment. The Disability of Arm-Shoulder-Hand score yielded an average of 41.5 points, which represents a moderate impairment of activities of daily living. Visual analog scale assessment demonstrated an overall high satisfaction (9.5 points; range, 1-10 points). The data demonstrate that even in severe necrotizing infections resulting in complex acute or chronic defects, limb salvage and infection control can be achieved successfully with radical debridement and early free tissue transfer. PMID- 12897518 TI - Use of the scrotal remnant as a tissue-expanding musculocutaneous flap for scrotal reconstruction in Paget's disease. AB - Wide excision of extramammary Paget's disease of the penoscrotal region may leave large defects that cannot be closed easily. The authors describe their experience with a series of 6 patients in whom reconstruction of the scrotal defect was undertaken using the scrotal remnant raised as a stretchable musculocutaneous flap. It was observed that as little as a third of the residual scrotum could be expanded to resurface the entire scrotum. All flaps survived completely. Severe scrotal edema and ecchymosis were observed in 1 patient but the symptoms resolved completely with Trendelenburg positioning. The penile defects were resurfaced individually with thick skin grafts. Good-quality take with no chordee was observed in all patients after initial reconstruction. One patient developed penile contracture after reexcision of recurrent disease. Mean follow-up was 22 months (range, 3-60 months). Large defects of as much as two thirds of the scrotum may be reconstructed successfully using the tissue-expanding scrotal musculocutaneous flap. PMID- 12897519 TI - Clinical, radiological, and audiological relationships in hemifacial microsomia. AB - Hemifacial microsomia is a craniofacial disorder involving structures derived from the first and second brachial arches. Bony structures, soft tissue, and the facial nerve can all be hypoplastic or absent. In this retrospective study of 70 patients at the Children's Hospital Los Angeles, craniofacial dysmorphology, temporal bone computed tomography, and audiological evaluations were examined. The purpose of this study was to identify further relationships between clinical findings, temporal bone anatomy, and audiological findings in these patients. Significant relationships were identified between total radiographic score and overall clinical findings (p<0.001). Clinical measurements of mandibular hypoplasia were also predictive of temporal computed tomographic findings (p<0.001), whereas clinical ear findings were related specifically to specific temporal bone abnormalities such as hypoplasia of the middle ear (p=0.008) and fusion or absence of ossicles (p<0.001). Interestingly, neither clinical findings nor temporal bone computed tomographic findings were related to either hearing loss type or degree. Audiology should not be deferred on the basis of relatively mild clinical findings. PMID- 12897520 TI - Predicting the risk of reoperation in metopic synostosis: a quantitative CT scan analysis. AB - Children with metopic synostosis have a well-described clinical picture of trigonocephaly, often with hypotelorbitism. The craniofacial disorder itself is well recognized; however, objective prognostic factors for predicting the risk of reoperation are not well known. In 39 children with metopic synostosis, measurements of the cranial length, cranial width, anterior intercoronal distance, anterior interorbital distance (intercanthal distance), lateral orbital distance, and interzygomatic buttress distance were taken from preoperative computed tomographic (CT) scans and were normalized relative to each child's age. To separate overall facial hypoplasia from regional hypoplasia, a ratio of intercanthal distance to interzygomatic buttress distance was determined. These prognostic factors were analyzed with respect to reoperation rate. A stepwise logistic regression analysis was used to determine the interrelationships between the prognostic factors. Twenty-eight percent of the children underwent reoperation (N=11, 1 total reoperation and 10 minor recontouring). All of the reoperations occurred in children with a decreased intercanthal distance (p=0.30). The ratio of intercanthal distance to midfacial width was related to reoperation rate, with those children who had a ratio < or =0.80 having a reoperation rate of 44% (8 of 18 total children with an intercanthal-to-zygomatic ratio < or =0.8, p=0.07). This relationship was significant in children younger than the age of 12 months (6 of 13 total children with an intercanthal-to zygomatic ratio < or =0.8, 46% reoperation rate, p=0.006). This study suggests that preoperative CT measurements can be used as a means of risk stratification in outcome analyses of the surgical treatment of craniosynostosis. In children treated for metopic suture synostosis, a foreshortened intercanthal distance compared with the interzygomatic buttress distance was related to reoperation rate, especially in children younger than 12 months of age. PMID- 12897521 TI - A review of emergency department fluid resuscitation of burn patients transferred to a regional, verified burn center. AB - The purpose of this study was to examine the adequacy of burn patient fluid resuscitation in relationship to the American Burn Association formula before arrival at a regional burn center. Further substratification of the data was undertaken to compare total burn surface area and fluid volume resuscitation as evaluated from the primary hospital's emergency department staff vs. the burn intensive care unit staff. The charts of all patients admitted to the burn center during 1 year were reviewed retrospectively. Data were compiled to calculate the time of injury, time of arrival in the referring emergency room, time in transit to the burn unit, and time of arrival in the burn unit. The total number of patients evaluated in the study was 41. Patients who were not referred from outside hospitals or who had incomplete charts were excluded. The average time from initial burn to transfer to the burn intensive care unit was 6.26 hours (range, 0.5-96 hours). The average total body surface area (TBSA) evaluated by the referring emergency department staff was 23.9% (range, 5-70%) compared with the burn intensive care unit staff evaluation average of 17.8% (range, 2-55%). Using the referring emergency department staff TBSA percentage, evaluation of the data revealed that only 23% of patients fell within the accepted range using the American Burn Association formula. Furthermore, 30% of patients were overresuscitated whereas 47% were underresuscitated. Of the overresuscitated patients, 1 patient was critically overresuscitated. In the group of underresuscitated patients, five were critically underresuscitated. Thirty-three percent of the patients' TBSA had a more than 50% discrepancy between the burn unit and the emergency department calculations. The authors conclude that better educating providers referring patients to regional burn centers can make a marked improvement in the overall care of burn patients. More important, early communication with the referring burn staff has been encouraged. Early communication permits review of estimated TBSA burn evaluations and permits cooperative calculations and optimal delivery of early fluid resuscitation. Burn center practitioners can improve care of patients before arrival by appropriately guiding the referring physician. PMID- 12897522 TI - Comparison of venous drainage in reverse-flow island flaps: an experimental study of the rabbit saphenous fasciocutaneous flap. AB - The mechanism of venous drainage in reverse-flow island flaps was investigated using the saphenous fasciocutaneous flap in New Zealand White rabbits. Ten animals were allocated into two groups of 10 flaps. In group I (left limb), the distal vascular pedicle (one saphenous artery and two venae comitantes) was not disturbed, maintaining the communicating and collateral branches intact. In group II (right limb), it was separated microsurgically from each other for 3 cm. Intravenous pressure was measured at 5, 15, 30, and 60 minutes after tourniquet release. The values of group II at 30 and 60 minutes were significantly lower (p<0.01) than those of group I. Ten days after flap elevation, the mean survival area of group I (95%) and group II (100%) was not significantly different. Histological examination of the vascular pedicle showed the saphenous veins in group II were more dilated than those of group I. These findings suggest that venous retrograde return in reverse-flow island flaps can be achieved more easily through a "direct incompetent valves route" than through a "circuitous communicating and collateral bypass route." PMID- 12897523 TI - Microcirculatory hemodynamics of the rat cremaster muscle flap in reduced blood flow states. AB - Recent studies revealed that muscle and musculocutaneous flaps have the lowest peripheral resistance and best flap survival. The critical values of flow rates responsible for this survival have not been established. The authors evaluated the effect of prolonged arterial flow reduction on flow hemodynamics and muscle flap survival, independent of neovascularization using the rat cremaster muscle island flap model for microcirculatory studies. Muscle flaps were implanted into a transparent Plexiglas chamber to allow prolonged observation of the microcirculation. Twenty-six male Sprague-Dawley rats were studied in three experimental groups. In group I (control; no occlusion), after flap isolation, the cremaster muscle was implanted into the tissue chamber and microcirculatory measurements were performed from day 0 to day 3. In group II, after flap isolation, flow in the main feeding artery (the iliac) was reduced with a silk suture loop between 47% and 62%. In group III, arterial flow was reduced between 63% and 80%. Vessel diameters, red blood cell velocities, number of perfused capillaries, and number of rolling, sticking, and transmigrating leukocytes were reduced daily for each rat. Immediately after flow reduction, mean arteriolar (A1) diameters were significantly smaller in group II (25%) and group III (29%) compared with the controls (p<0.05). This reduction was confirmed by a drop in red blood cell velocities of 37% and 58% in groups II and III respectively (p<0.05). At day 1, more than 60% of group III flaps had no arterial flow, whereas normal flow was observed in control flaps for as long as 3 days. In addition, the number of perfused capillaries dropped significantly (by 42%) in group II flaps after 3 days (p<0.05). The authors conclude that continuous arterial blood flow reduction exceeding 60% of the normal volume, independent of angiogenesis, is incompatible with flap viability in the rat cremaster muscle model. PMID- 12897524 TI - Suction lipectomy during flap reconstruction provides immediate and safe debulking of the skin island. AB - Immediate suction lipectomy combined with flap reconstruction has not been reported to date. Concern of skin island compromise has swayed clinicians from attempting synchronous debulking of generous skin island tissue during primary flap reconstruction. To date, several reports have documented the safe use of liposuction during mobilization of skin flaps as a minimally invasive, undermining tool in the cervical neck area. Additionally, liposuction has been used safely as an adjunct to rhytidectomy procedures over areas containing vital structures. The authors report 2 patients in whom generous skin island tissue prevented primary closure after transposition of the flap into its recipient site. Aspiration of subcutaneous tissue in a transverse rectus abdominis musculocutaneous flap and gluteus musculocutaneous rotational flap for breast and sacral ulcer reconstructions respectively resulted in appropriate debulking of the skin island without any flap skin compromise. The findings of this report may provide impetus for further clinical investigations establishing the utility of combined suction lipectomy and flap reconstruction. PMID- 12897525 TI - Free perforator flap for the treatment of defects after resection of huge arteriovenous malformations in the head and neck regions. AB - The authors report two cases of huge arteriovenous malformations in the head and neck regions treated successfully with preoperative superselective transarterial embolization and resection followed by a free perforator flap transfer. Based on the authors' previous cases, en block mass resection of the malformation was possible with bleeding of less than 150 ml. The massive defects could be repaired with free perforator flaps using an anterolateral thigh flap and a deep inferior epigastric artery perforator flap. One patient who lost facial muscle underwent reconstruction by simultaneous muscle transfer, and both patients regained acceptable cosmetic appearance and dynamic facial function. Now, more than 4 to 7 years after surgery, the patients have shown no reexpansion of the malformation. The important points of this treatment are complete embolization to accomplish total resection with minimal bleeding, free flap transfer to prevent postoperative reexpansion or recurrence of arteriovenous malformations, and the selection of recipient vessels because of arterial embolization in part of the lesion. PMID- 12897526 TI - Free gluteal artery perforator flap with a short, small perforator. AB - The introduction of supermicrosurgery has led to the development of a new gluteal perforator flap nourished only by a musculocutaneous perforator of the superficial gluteal artery system. This flap has a perforator that is short (3-4 cm in length) and small (less than 1 mm). The successful transference of a free gluteal perforator flap for the coverage of soft-tissue defects in the foot and face in two patients is described in this article. With this flap, deeper and longer dissection for a pedicle vessel is unnecessary, flap elevation time is shorter, thinning of the flap with primary defatting and creation of an adiposal flap with customized thickness for tissue augmentation are possible, the donor site is in a concealed area with minimal donor site morbidity, and application of the flap as a scarred flap for previous operations in the gluteal region is possible. The disadvantages of this flap are necessity of dissection for a smaller perforator and of anastomosis of small-caliber vessels of less than 1.0 mm. PMID- 12897527 TI - Double V-Y advancement flap in the management of lower lip reconstruction. AB - A new method for lower lip reconstruction is proposed. It is based on a double cutaneous and mucous V-Y advancement flap and is suggested for the surgical treatment of lip cancer confined between the skin and the vermilion with a horizontal spread. Sixteen patients were treated from 1996 to 2001 with this technique and the results were satisfactory both from the functional and the aesthetic standpoints. PMID- 12897528 TI - A review of mechanical adjuncts in wound healing: hydrotherapy, ultrasound, negative pressure therapy, hyperbaric oxygen, and electrostimulation. AB - Chronic or non-healing wounds may develop in the setting of many diseases and are the source of considerable morbidity as well as health costs. These wounds demand an aggressive, multifactorial approach including surgical debridement, revascularization, antibiotics and dressings. In addition several adjuvant treatment methods have been developed to further stimulate healing. Whirlpool, although used frequently, has not been proven to be of benefit. However, pulsed lavage does show a promising future. Ultrasound has demonstrated beneficial effects but further controlled studies are needed. Subatmospheric pressure therapy is associated with few complications and is fast becoming a mainstay of adjuvant therapy. Hyperbaric oxygen therapy has been shown to be effective for many types of wounds. Unfortunately, cost and access to chambers may prohibit its use on a routine basis. Finally, electrostimulation may be one of the up and coming therapies for the future. Though, more studies are needed to determine the mode of delivery for various types of wounds. PMID- 12897529 TI - The prince of pinnae. PMID- 12897530 TI - Supercharged TRAM flaps can be a worthwhile procedure! PMID- 12897531 TI - Re: vulvoperineal reconstruction. PMID- 12897532 TI - Re: modification of split-thickness skin graft. PMID- 12897533 TI - Ligated punctured expanders: are they clinically safe? PMID- 12897534 TI - Upper lip orocutaneous fistula in edentulous maxilla. PMID- 12897535 TI - Use of silicone block as a pellet in the treatment of minor malocclusion. PMID- 12897536 TI - Validating competency at the bedside. AB - Nurses' ability to practice is constantly challenged by changes in equipment, staffing, acuity, time constraints, and requirements of regulatory agencies. This article highlights an innovative approach to validate competency of registered nurses in a 380-bed hospital in the Midwest. Confronted with increasing demands to validate competency on high-priority issues, as well as in areas of high risk, educators developed a plan to verify competency by using an observed performance format. Although the specific competency was pain management, this prototype can be used for validation of other high-risk or high-profile competencies. PMID- 12897537 TI - Role acquisition, socialization, and retention: unique aspects of a mentoring program. AB - Banner Good Samaritan Medical Center, a level-one tertiary care teaching facility in the southwest United States, developed an 18-month RN mentoring program in response to the national nursing shortage and a desire to promote nursing retention at the institution. PMID- 12897538 TI - Standing in their shoes: an experiential learning experience in promoting age sensitivity. AB - The anticipated rise in the elderly population has been well documented. Nurses working in staff development have an important role in enabling healthcare staff to gain insights into the challenges of the aging process and positively affect the quality of life of elderly clients. This experiential program incorporates a variety of learning strategies to immerse staff in the elderly clients' world. Evaluations of the session have identified the need for staff to exercise patience, be proactive in anticipating elderly clients' needs, and appreciate client individuality. PMID- 12897539 TI - Project management: a core competency for professional nurses and nurse managers. AB - Project management is a core competency for nurses and managers. In this article, the project life cycle and project planning and controlling tools are described. Managers can readily see that using project management can be done with minimal training and disruption to the organization. Clearly, the costs of adopting project management are small compared to the benefits in productivity, reduced work stress, and reduced interpersonal conflict. PMID- 12897541 TI - I CARE: an organization-wide customer service education program. AB - When a survey indicated that patients recently discharged from The Brant Community Healthcare System (BCHS) believed that customer service across the organization could be enhanced, planning to enhance the quality of service for both internal and external customers began immediately. This article describes the measures taken to realize this improvement. It describes the cultural change required, important things to consider when planning customer service programs in healthcare settings, and some of the content of the customer service education program that was ultimately delivered to leaders, staff, physicians, and volunteers throughout the organization. PMID- 12897542 TI - A charge nurse workshop based on research. AB - The authors describe a charge nurse workshop based on a qualitative research study of charge nurse competencies and a commitment to include all levels of staff in the planning process. The workshop met the needs of nursing staff to have an interactive, educational experience that addressed leadership, human relations, and team-building skills needed for the charge nurse role and was fun. Evaluation of the workshop with lessons learned also is presented. PMID- 12897543 TI - Instant teaching tools. PMID- 12897545 TI - Hypothermia during head and neck surgery. AB - OBJECTIVE: To determine the predictors and incidence of hypothermia in patients undergoing head and neck surgery. STUDY DESIGN: Retrospective analysis. METHODS: Patients were either not warmed (n = 43) or actively warmed with forced-air warming (n = 25). Clinical variables that were assessed as predictors of core body temperature included age, body mass, duration of procedure, estimated blood loss, amount of intravenous fluids administered, and the use of forced-air warming. The incidence of severe intraoperative hypothermia and potential hypothermia-related complications was also examined. RESULTS: The study demonstrated that advanced age is a risk factor for hypothermia and decreased body mass is associated with lower final body temperatures in the groups of patients that was not warmed. After adjusting for differences in the ages and weights between the two groups, the mean core body temperature was found to be 0.4 degrees C lower in the patients who were not warmed. Severe intraoperative hypothermia occurred in 5 of 38 patients (11.6%) who were not warmed and 2 of 23 patients (8.0%) who were warmed. The complications associated with hypothermia included delayed time to extubation, the development of neck seromas, and flap dehiscence. CONCLUSIONS: Patients undergoing head and neck surgery are at risk for the development of intraoperative hypothermia and require careful temperature monitoring. Elderly patients and patients with low body mass are more prone to develop low intraoperative core body temperatures. Active warming with forced-air warmers should be considered for patients at risk for intraoperative hypothermia and for patients who develop hypothermia intraoperatively, to avoid hypothermia related complications. PMID- 12897546 TI - Incidence, etiology, and management of cerebrospinal fluid leaks following trans sphenoidal surgery. AB - OBJECTIVES/HYPOTHESIS: The incidence of cerebrospinal fluid (CSF) leak following trans-sphenoidal surgery ranges from 0.5% to 15.0%. Factors predicting which patients are likely to develop postoperative leaks and optimal management of these patients are poorly defined. The objective was to determine 1) the incidence of CSF leak following trans-sphenoidal surgery; 2) demographic or intraoperative factors associated with postoperative leaks; 3) techniques and efficacy of postoperative leak management at Oregon Health and Science University, (Portland, OR). STUDY DESIGN: Retrospective chart review. METHODS: Two hundred thirty-five trans-sphenoidal surgeries were performed on 216 patients between 1994 and 2001. Follow-up data were available for 217 operations (92.3%) performed on 202 patients (93.5%). RESULTS: Postoperative CSF leaks occurred in 6.0% (13 of 217) of patients. Leaks were more common in the setting of revision surgery versus primary surgery (14.6% vs. 4.0%, P =.0096), nonadenomatous disease versus pituitary adenoma (15.8% vs. 5.1%, P =.059), or if an intraoperative leak occurred (12.7% vs. 2.7%, P =.004). However, on multivariate analysis, only the presence of intraoperative leak (P =.008) and nonadenomatous disease (P =.047) were found to be independently associated with postoperative CSF leak. Size of adenoma was not associated with occurrence of postoperative CSF leak (6.4% for microadenoma vs. 4.2% for macroadenoma) on both univariate and multivariate analysis. There were 13 postoperative CSF leaks: 2 resolved with lumbar drainage and 11 required operative management (three required multiple procedures). Of the three patients who required multiple operations, two had hospital courses complicated by meningitis and postinfectious hydrocephalus and ultimately required ventriculoperitoneal shunts. Endoscopic re-exploration was successful in 87.5% (7 of 8) of cases. CONCLUSIONS: Cerebrospinal fluid leaks following trans sphenoidal surgery occurred in 6.0% of cases. Nonadenomatous disease and presence of an intraoperative leak were independent predictors of a postoperative leak. Endoscopic re-exploration combined with packing was an effective technique in managing uncomplicated postoperative leaks. In the setting of meningitis and postinfectious hydrocephalus, more invasive techniques such as ventriculoperitoneal shunt may be necessary. PMID- 12897547 TI - Dermatofibrosarcoma protuberans of the head and neck: treatment with Mohs surgery using inverted horizontal paraffin sections. AB - OBJECTIVE: To describe results of treating dermatofibrosarcoma protuberans of the head and neck with a Mohs surgical technique in which tumor margins were evaluated with inverted horizontal paraffin sections to identify the margin of resection necessary to achieve tumor-free borders. STUDY DESIGN: Retrospective case study of nine patients. METHODS: In each case, Mohs surgery using inverted horizontal frozen-section analysis was continued until tumor-free margins were achieved. The tumor-free specimens collected during this procedure were immediately reprocessed by using inverted horizontal paraffin sectioning, and the histological results of the two techniques were compared. To identify the margins of resection needed to achieve tumor-free margins, preoperative lesion size was compared with postoperative defect size. RESULTS: Paraffin section analysis showed persistent tumor in seven of the nine patients in whom the frozen-section technique was used to clear the tumor. Tumor-free margins were achieved in all patients by using one additional excision. The tumor had spread extensively beyond clinical margins and required a mean minimum margin of 3.7 cm around the gross area of tumor (margin range, 2.5-6.0 cm). Patients treated by this technique showed no local, regional, or distant recurrence at follow-up performed at a mean time of 43 months postoperatively (range, 19-74 mo). CONCLUSIONS: Dermatofibrosarcoma protuberans of the head and neck is a tumor that shows extensive infiltration beyond gross margins. Wide local excision with 2- to 3-cm margins results in an unacceptably high recurrence rate; larger excisional margins are necessary to remove all disease. Accurate margin identification achieved by using Mohs surgery with rush paraffin section analysis results in complete tumor removal and excellent control rates. This method should be preferred for treatment of dermatofibrosarcoma protuberans of the head and neck. PMID- 12897548 TI - Swallowing function and tracheotomy dependence after combined-modality treatment including free tissue transfer for advanced-stage oropharyngeal cancer. AB - OBJECTIVES/HYPOTHESIS: There are many treatments available for advanced oropharyngeal cancer. Organ-sparing protocols reserve surgery for salvage and are thought to provide adequate rehabilitation. Surgical resection with free tissue transfer may also provide adequate functional rehabilitation. The objective was to describe swallowing status and time to decannulation in a series of patients treated with combined-modality therapy that included free flap reconstruction. STUDY DESIGN: Retrospective chart review. METHODS: Patient data were obtained from medical records of 20 patients with stage III or IV oropharyngeal carcinoma, who were consecutively treated with surgical tumor extirpation, free flap reconstruction, and postoperative irradiation at a tertiary academic center from 1985 to 2002. The following variables were identified: patient and tumor characteristics, free flap type, irradiation data, and airway and swallowing status before and after treatment. RESULTS: One patient underwent total laryngopharyngectomy, and the remaining 19 patients underwent tracheotomy at the time of definitive surgery. Free flap reconstructions included 1 ulnar and 15 radial forearm fasciocutaneous flaps and 4 fibula osteocutaneous flaps. Postoperatively, all 19 tracheotomized patients had successful decannulation. Average time to decannulation was 15 days (range, 3-42 d). After surgery and before irradiation, 13 patients initiated oral intake, on average, at 19.5 days (range, 7-28 d); 6 patients required no additional supplementation. By 4 months after surgery, having completed radiation therapy, 10 patients were consuming all nutrition orally; the other 10 patients still required tube-feed supplementation, although 6 of these patients were also eating by mouth. CONCLUSION: Combined modality treatment that includes free flap reconstruction for advanced-stage oropharyngeal cancer may provide reasonable functional rehabilitation with respect to postoperative airway and swallowing. PMID- 12897549 TI - The impact of malignant disease on facial nerve function after parotidectomy. AB - OBJECTIVES/HYPOTHESIS: Studies of immediate postoperative facial nerve function following parotidectomy focus on benign disease. The purpose of the study was to compare facial nerve function with regard to benign or malignant disease in patients undergoing superficial parotidectomy. STUDY DESIGN: Retrospective cohort study of consecutive patients undergoing parotidectomy between 1995 to 2002. METHODS: House-Brackmann (HB) grade was recorded or assigned at the first postoperative visit. For patients with HB grade of III or greater, final resolution of facial nerve function was recorded. A chi2 analysis for independence was conducted between patients with HB grade of II or less and those with HB grade of III or greater and final pathological finding (benign vs. malignant disease). Mean and median times to resolution were determined for patients with HB grade of III or greater. Times to resolution for benign and malignant disease for those with HB grade of III or greater were compared (Kaplan Meier method). RESULTS: Sixty-seven patients with benign disease and 52 with malignant disease were included. Ninety-four percent of patients with benign disease had HB grade of II or less at first postoperative visit (mean period, 11.6 d) compared with 76.9% of patients with malignant disease (mean period, 12.4 d). A chi2 analysis found this difference significant (chi2 = 7.36, P =.0067). Time to resolution for HB grade of III or greater was 253.8 days (+/-168.2 d) with median time of 229.5 days for benign disease and 182.4 days (+/-134.6 d) with median time of 138 days for malignant disease. Kaplan-Meier comparison found no significant differences in time to final resolution (P =.83). Three patients in the group with malignant disease had unresolved dysfunction (one patient for >2 y). CONCLUSIONS: Patients with benign disease have a greater chance of having HB grade of II or less immediately following surgery; however, whether the disease is benign or malignant, long-term final facial nerve function is the same. PMID- 12897550 TI - Subglottic stenosis associated with Wegener's granulomatosis. AB - OBJECTIVE: To evaluate the outcomes of subglottic stenosis in patients with Wegener's granulomatosis. STUDY DESIGN: Retrospective chart review. METHODS: A retrospective chart review was undertaken to review all patients evaluated in an otolaryngology-head and neck surgery department between 1993 and 2001 for active symptoms attributed to subglottic stenosis who had undergone cytoplasmic staining antineutrophil cytoplasm antibody and perninuclear staining antineutrophil cytoplasm antibody testing and who had a clinical diagnosis of Wegener's granulomatosis. RESULTS: Twenty-seven patients were identified with Wegener's granulomatosis and subglottic stenosis. Among these, 11 of 27 (40.7%) underwent tracheotomy; 13 of 27 (48.1%) required multiple surgical procedures. A history of open laryngotracheal repair was present in 7 of 27 patients (25.9%). CO(2) laser resection and dilation was used in 12 of 27 patients (44.4%). Three of 27 patients (11.1%) had a tracheotomy at last follow-up. Patients were followed for an average duration of 76.8 months (6.4 y) from the onset of symptoms attributed to subglottic stenosis. All patients who underwent open laryngotracheal reconstruction had successful decannulation. CONCLUSIONS: Wegener's granulomatosis is a necrotizing vasculitis that can involve the subglottis. Our approach to airway management in these patients involves a tracheotomy if obstruction occurs during acute inflammatory activity. We recommend minimizing airway manipulation during episodes of systemic disease activity. Once control of the disease process is obtained, consideration is given to either an endoscopic or an open approach based on the extent of stenosis. Either laryngotracheal reconstruction or CO(2) laser resection and dilation may be undertaken with safety and should result in favorable outcomes. PMID- 12897551 TI - Tacrolimus and mycophenolate mofetil provide effective immunosuppression in rat laryngeal transplantation. AB - OBJECTIVES/HYPOTHESIS: Tacrolimus is efficacious in several transplantation settings. Some studies have demonstrated improved results using combination therapy with mycophenolate mofetil. Our primary objective was to evaluate the efficacy and optimal dosing of tacrolimus in preventing rejection, using an established rat model of laryngeal transplantation. Further, the ability of mycophenolate to allow lower dosing of tacrolimus while achieving equivalent immunosuppression was investigated. STUDY DESIGN: A dosage efficacy study with 10 experimental arms was conducted. METHODS: Dosage groups were 0.1, 0.2, 0.3, and 0.6 mg/kg tacrolimus alone and 0.1 mg/kg tacrolimus combined with 15 mg/kg mycophenolate mofetil, 0.1 mg/kg tacrolimus combined with 30 mg/kg mycophenolate mofetil, 0.1 mg/kg tacrolimus combined with 40 mg/kg mycophenolate mofetil, 0.2 mg/kg tacrolimus combined with 15 mg/kg mycophenolate mofetil, 0.2 mg/kg tacrolimus combined with 30 mg/kg mycophenolate mofetil (30 d only), and 0.2 mg/kg tacrolimus combined with 40 mg/kg mycophenolate mofetil. Each group contained 8 to 10 rats. Grafts were harvested for histopathological analysis on day 15 or 30 after transplantation. Histopathological appearance of the graft was blindly graded according to an established scale. Dosage groups were compared on rejection score using Wilcoxon's rank sum test and the Jonckheere-Terpstra test for trend. RESULTS: There was a significant association between increasing dose of tacrolimus and decreasing rejection score at both 15 and 30 days (P <.001). In the groups treated with 0.1 mg/kg T, an increasing dose of mycophenolate was associated with lower rejection scores at both 15 and 30 days (P =.001). In the group treated with 0.2 mg/kg T, there was no evidence that the addition of mycophenolate resulted in lower rejection at 15 days. However, at 30 days, combination therapy with increasing doses of mycophenolate was associated with decreasing rejection score (P =.002). CONCLUSIONS: Tacrolimus is an effective immunosuppressive agent for laryngeal transplantation. Mycophenolate mofetil allows lower doses of tacrolimus to be used while preserving graft viability in the early post-transplantation period. PMID- 12897552 TI - Complete laryngotracheoesophageal cleft: complicated management issues. AB - OBJECTIVES/HYPOTHESIS: Laryngotracheoesophageal clefts are rare developmental anomalies of the upper aerodigestive tract. They range in severity from being virtually asymptomatic throughout life (type I) to being incompatible with life (type IV). The timing of diagnosis is crucial to the successful treatment of severe clefts. Treatment is complicated and requires a multi-team approach. The case report discusses the elements involved in diagnosing and treating this severe airway anomaly. STUDY DESIGN: Case report. METHODS: Review of a case at a tertiary care center. RESULTS: A case of a complete laryngotracheoesophageal cleft with left-side pulmonary agenesis in a newborn is reported. Rigid bronchoscopy revealed a common tracheoesophageal lumen from the larynx to the stomach with a single bronchus supplying the right-side lung. Management of this patient included establishment and maintenance of a tenuous airway, maintenance of nutrition, and anesthetic and surgical planning for upper aerodigestive tract reconstruction. CONCLUSIONS: Although severe laryngotracheoesophageal clefts are rare, they require prompt, team-oriented management for the best outcome possible. The diagnosis, sustenance, and treatment options of these patients depend on varied and complicated factors, which are discussed. PMID- 12897553 TI - Serum antidiuretic hormone levels in patients with unilateral Meniere's disease. AB - OBJECTIVES/HYPOTHESIS: Prior studies have indicated a possible role of antidiuretic hormone (ADH) in the pathogenesis of Meniere's disease. Animal studies have shown presence of ADH receptors in the inner ear, and chronic vasopressin administration has been shown to induce endolymphatic hydrops. Furthermore, elevation of serum ADH levels in human has been noted in patients with Meniere's disease. The goal of the study report was to analyze ADH levels in a series of patients with definite unilateral Meniere's disease to further investigate this relationship. STUDY DESIGN: Retrospective analysis METHODS: Antidiuretic hormone levels were obtained from 26 consecutive patients with a diagnosis of definite Meniere's disease as defined by the 1995 guidelines from the Committee of Hearing and Equilibrium (American Academy of Otolaryngology-Head and Neck Surgery). The ADH levels were drawn for each patient within one week of an acute episode of vertigo. These values were compared to ADH levels from 31 healthy volunteers. Statistical analysis was performed using a two-tailed t test. RESULTS: Mean ADH level for patients with Meniere's disease was 4.07 pg/mL (SD = 2.82 pg/mL) and for the control group, 3.37 pg/mL (SD = 1.48 pg/mL). The difference in ADH levels was not statistically significant (P >.05). CONCLUSIONS: Although previous reports have demonstrated a possible role of ADH in the pathogenesis of Meniere's disease, the study did not show a statistically significant elevation of ADH levels in patients with unilateral Meniere's disease. Before excluding an ADH-inner ear pathogenic relationship, ADH levels in patients with bilateral Meniere's disease should be investigated. PMID- 12897554 TI - The superior petrosal triangle as a constant anatomical landmark for subtemporal middle fossa orientation. AB - OBJECTIVES/HYPOTHESIS; Anatomical landmarks including the arcuate eminence and the superficial petrosal nerve serve as orienting landmarks for middle fossa dissection. However, because of considerable variation among patients, these landmarks are not always readily identifiable. We expand on a previously described method for identifying the head of the malleus as a constant anatomical landmark to optimize exposure when employing a middle fossa approach. METHODS; We completed an anatomical study using 10 preserved human cadaveric temporal bones to define the anatomical relationship among the root of the zygoma, the posterior lateral lip of the foramen spinosum, and the bony tegmen over the head of the malleus. Subsequently, 5 fresh whole human cadaveric heads (10 temporal bones) were dissected using a surgically oriented anterior petrosectomy-middle fossa approach to evaluate the consistency of localizing the head of the malleus. RESULTS; We defined the superior petrosal triangle as a stable anatomical relationship. Our cadaveric data demonstrated that the distance from the root of the zygoma to the head of the malleus was 18.7 mm (SD = 1.7 mm) and the distance from the foramen spinosum to the head of the malleus was 19.2 mm (SD = 1.0 mm). The intersection of an arc transcribed 19 mm from the root of the zygoma and an arc transcribed 19 mm from the foramen spinosum localized the head of the malleus within 2.5 mm (SD = 2.4 mm). CONCLUSIONS: The landmarks defined by the superior petrosal triangle represent a means to localize the bony tegmen over the head of the malleus. Identification of the head of the malleus as a landmark in middle fossa surgery when other landmarks are not recognizable optimizes patient safety and surgeon confidence during complex surgical procedures. PMID- 12897555 TI - Surgical excision of acoustic neuroma: patient outcome and provider caseload. AB - OBJECTIVES/HYPOTHESIS: For many complex surgical procedures, larger hospital or surgeon caseload is associated with better patient outcome. We examined the volume-outcome relationship for surgical excision of acoustic neuromas. STUDY DESIGN: Retrospective cohort study. METHODS: The Nationwide Inpatient Sample (1996 to 2000) was used. Multivariate regression analyses were adjusted for age, sex, race, payer, geographic region, procedure timing, admission type and source, medical comorbidities, and neurofibromatosis status. RESULTS: At 265 hospitals, 2643 operations were performed by 352 identified primary surgeons. Outcome was measured on a four-level scale at hospital discharge: death (0.5%) and discharge to long-term care (1.2%), to short-term rehabilitation (4.4%), and directly to home (94%). Outcomes were significantly better after surgery at higher-volume hospitals (OR 0.47 for fivefold-larger caseload, P <.001) or by higher-volume surgeons (OR 0.46, P <.001). Of patients who had surgery at lowest-volume quartile hospitals, 12.3% were not discharged directly home, compared with 4.1% at highest-volume-quartile hospitals. There was a trend toward lower mortality for higher-volume hospitals (P =.1) and surgeons (P =.06). Of patients who had surgery at lowest-caseload-quartile hospitals, 1.1% died, compared with 0.6% at highest-volume-quartile hospitals. Postoperative complications (including neurological complications, mechanical ventilation, facial palsy, and transfusion) were less likely with high-volume hospitals and surgeons. Length of stay was significantly shorter with high-volume hospitals (P =.01) and surgeons (P =.009). Hospital charges were lower for high-volume hospitals (by 6% [P =.006]) and surgeons (by 6% [P =.09]). CONCLUSION: For acoustic neuroma excision, higher-volume hospitals and surgeons provided superior short-term outcomes with shorter lengths of stay and lower charges. PMID- 12897556 TI - Facial nerve paralysis following cochlear implant surgery. AB - OBJECTIVES: Facial nerve paralysis is a rare but devastating complication of cochlear implant surgery. The aims of the study were to define the incidence of facial nerve paralysis in our series and understand possible mechanisms of injury. STUDY DESIGN: Retrospective chart review and case reports. METHODS: Charts were reviewed of all 705 patients implanted between 1980 and 2002 at the authors' institutions to identify those with postoperative facial nerve weakness and determine incidence. For patients with facial nerve weakness, onset, degree, and timing of paralysis were noted; clinical findings were correlated to operative report findings. The method of treatment was noted, and the final facial nerve function outcome was recorded. RESULTS: Five patients (one child and four adults) were found to have postoperative facial nerve weakness, for an incidence of 0.71%. This complication was delayed in all cases, ranging from 18 hours to 19 days postoperatively. All patients were treated with steroids or steroids combined with antiviral medication, and all ultimately recovered normal facial function. CONCLUSIONS: In the study series, the incidence of facial nerve paralysis following cochlear implant surgery was 0.71%. Possible mechanisms of injury included heating injury and viral reactivation. All patients presented with a delayed facial nerve paralysis and did recover normal facial nerve function. PMID- 12897557 TI - Alpha 1-antitrypsin and ilomastat inhibit inflammatory proteases present in human middle ear effusions. AB - OBJECTIVES: Proteases of both the serine and metalloproteinase families have been shown to play a role in the pathogenesis of otitis media (OM). Inhibitors of proteases from each of these families have been shown to beneficially impact disease progression in a number of related chronic inflammatory conditions, but their use has not been studied in OM. The purpose of this study was to assess the activity of the protease inhibitors recombinant alpha 1-antitrypsin (rAAT) and ilomastat on inflammatory proteases present in human middle ear effusions (MEEs), with a view to their potential utility in the treatment of OM. STUDY DESIGN: Prospective and ex vivo. METHODS: MEEs were collected from 100 patients presenting for middle ear surgery, most commonly tympanostomy tube placement or treatment of acute posttympanostomy otorrhea (APTO). MEEs were analyzed for the presence of matrix metalloproteinases (MMPs) and human neutrophil elastase (HNE) and the inhibitory activity of rAAT and ilomastat on these proteases, respectively. RESULTS: MMP levels were highest in APTO, and HNE was highest in chronic suppurative OM and APTO. High levels of MMP and HNE (>3 mAU/min) were found in 52% and 37% of MEEs, respectively. Ilomastat and rAAT demonstrated significant inhibition of MMP and HNE activity (>30% reduction), respectively, in 80% and 82% of MEEs with high levels of activity. CONCLUSIONS: Proteases are commonly found in OM. Ilomastat and rAAT are potent inhibitors of proteases in MEEs across a wide range of OM in humans. Investigation into the potential therapeutic benefits of these protease inhibitors is warranted. PMID- 12897558 TI - A new method for closing tympanic membrane perforations using basic fibroblast growth factor. AB - OBJECTIVE: To present a new method for closing tympanic membrane perforations using basic fibroblast growth factor (bFGF) combined with an atelocollagen/silicone bilayer membrane as a patch material. STUDY DESIGN: Closure of tympanic membrane perforations was attempted using bFGF, which is thought to facilitate the growth of fibroblasts and collagen fibers at the margin of the perforation. METHODS: Under an operating microscope, the margin of the perforation was trimmed, and a piece of an atelocollagen/silicone bilayer membrane infiltrated with 0.2 mL Trafermin (0.1% solution) (bFGF group) or saline (control group) was then placed in the perforation with the silicon layer facing outward. Nine patients were treated with bFGF, and five were treated with saline. Data obtained from patient records included patient age, perforation size, and duration of treatment, with a focus on hearing improvement and complete tympanic membrane closure. RESULTS: The mean perforation size before treatment was 16.5% in the bFGF group and 9.6% in the control group. Closure of the tympanic membrane perforation was achieved in all cases in the bFGF group, whereas it was achieved in only two of five cases in the control group. With bFGF treatment, the tympanic membrane perforations closed completely within 3.7 weeks, and hearing improved by 13.3 dB in the bFGF group. CONCLUSION: The study demonstrated that bFGF combined with an atelocollagen/silicone bilayer membrane is effective for the conservative treatment of tympanic membrane perforation. PMID- 12897559 TI - Embolic inner ear decompression illness: correlation with a right-to-left shunt. AB - OBJECTIVES/HYPOTHESIS: Inner ear decompression illness is thought to be a rare phenomenon in recreational divers, isolated signs and symptoms of inner ear dysfunction usually being attributed to inner ear barotrauma. STUDY DESIGN: We present 11 cases of inner ear dysfunction in nine divers with inner ear decompression illness. RESULTS: All nine divers had significant right-to-left shunt as diagnosed by transcranial Doppler sonography. CONCLUSIONS: The authors thought that mechanism of causation in these cases may have been intravascular bubble emboli and that inner ear decompression illness may be more common among recreational divers than currently recognized. Failure to treat inner ear decompression illness with recompression therapy can result in permanent disability. Because the differential diagnosis between inner ear barotrauma and inner ear decompression illness can be impossible, the authors suggested that divers who present with inner ear symptoms following a dive should have recompression immediately after having undergone bilateral paracentesis. PMID- 12897560 TI - Temporal bone histopathological and quantitative analysis of mitochondrial DNA in MELAS. AB - OBJECTIVES/HYPOTHESIS: Although hearing loss is common in MELAS (syndrome of mitochondrial encephalopathy, lactic acidosis and stroke-like episodes), the histopathology of the temporal bone has not been reported. The majority of cases of MELAS are linked to a mitochondrial DNA (mtDNA) mutation at nucleotide 3243. In MELAS, normal mtDNA and mutant mtDNA coexist in a heteroplasmic manner. The purpose of the study was to report the otopathological findings from two patients with MELAS and quantitative mtDNA analysis in the inner ear of one of these patients. STUDY DESIGN: Basic scientific histopathological examination and quantitative mtDNA analysis of the temporal bone. METHODS: Temporal bones were embedded in celloidin and sectioned for light microscopic study. Graphic reconstruction of the cochlea was performed using the method described by Schuknecht. For quantitative mtDNA analysis, total DNA from the membranous part of the inner ear was collected, amplified by polymerase chain reaction, and digested with the restriction enzyme. The percentage of mutant/total mtDNA was measured by the ratio of fluorescence intensity. RESULTS: Histopathological examination revealed severe degeneration of the stria vascularis and degenerative change of spiral ganglion cells in both patients. The quantitative DNA studies showed that the proportion of mutant to wild-type mtDNA was similar in both histologically affected and histologically unaffected tissues within the inner ear. CONCLUSION: Dysfunction of the stria vascularis and spiral ganglion cells causes sensorineural hearing loss in MELAS. PMID- 12897561 TI - Intravenous anesthesia provides optimal surgical conditions during microscopic and endoscopic sinus surgery. AB - OBJECTIVES/HYPOTHESIS: Controlled hypotension is used to improve surgical conditions during microscopic and endoscopic sinus surgery. New short-acting anesthetics such as propofol and remifentanil allow exact control of intraoperative blood pressure and thus might be valuable tools to improve intraoperative conditions for the otorhinolaryngological surgeon. Intravenous anesthesia was compared with traditional balanced anesthesia by subjective assessment of surgical conditions made by two experienced otorhinolaryngological surgeons. STUDY DESIGN: Prospective, randomized, patient- and observer-blinded study. METHODS: Ninety consecutive patients were randomly assigned to receive intravenous anesthesia with propofol 5 to 8 mg.kg-1.h-1 and remifentanil 10 to 30 microg.kg-1.h-1 or with isoflurane (0.4-1.0 vol%) and repetitive doses of 0.5 to 1 mg alfentanil. An injectable vasodilator was used in both groups to keep mean arterial pressure between 60 and 70 mm Hg (8-9.3 hecto-pascal). The attending otorhinolaryngological surgeon was unaware of the type of anesthesia administered. Immediately after the operation the surgeons rated surgical conditions (bleeding from the surgical field) on a visual analogue scale (0-10 cm) and on a verbal rating scale. RESULTS: Blood pressure was not different between the two groups, but heart rate was lower in the intravenous anesthesia group (mean heart rate in the intravenous anesthesia group, 62 beats per min [95% confidence interval, 52-72]; mean heart rate in the balanced anesthesia group, 75 beats per min [95% confidence interval, 67-83]). Surgical conditions were rated to be significantly better (P <.0001) during anesthesia with propofol remifentanil (median rating: 2.8; 25th/75th percentile: 2.0/3.4) compared with isoflurane-alfentanil (median rating: 4.9; 25th/75th percentile: 3.6/7.6). CONCLUSIONS: Intravenous anesthesia using propofol-remifentanil provides better surgical conditions compared with a traditional balanced anesthesia technique using isoflurane-alfentanil. It is hypothesized that lower cardiac output caused by decreased heart rate during deep general anesthesia is responsible for this result. PMID- 12897562 TI - The effect of nasal polyp epithelial cells on eosinophil activation. AB - OBJECTIVES/HYPOTHESIS: Eosinophil infiltration into an inflammatory site is a characteristic histological finding in patients with chronic rhinosinusitis and nasal polyps. Most of the eosinophils in chronic rhinosinusitis are activated in the nasal cavity, but the exact activation mechanism of eosinophils is unknown. The study was designed to investigate the effect of human nasal epithelial cells on the activation of eosinophils. STUDY DESIGN: Peripheral blood eosinophils were isolated from healthy volunteers and incubated in human nasal polyp epithelial cell conditioned media (HPECM). Superoxide production and eosinophil-derived neurotoxin were measured to determine eosinophils activation. HPECMs were assayed by ELISAs for interleukin-8 (IL-8), granulocyte-macrophage colony stimulating factor (GM-CSF), eotaxin, and regulated on activation normal T expressed and secreted (RANTES). To identify the chemical mediators involved in the activation of eosinophils. RESULTS: HPECM (n = 7) contained 31.48 ng/mL interleukin-8, 533.43 pg/mL GM-CSF, 5.90 pg/mL eotaxin, and 11.06 pg/mL RANTES. Eosinophils were activated by HPECM and inhibited only by anti-GM-CSF antibody, not by the other chemical mediators. CONCLUSION: The results suggest that eosinophils in nasal secretions are activated by GM-CSF, which is produced by nasal epithelial cells. PMID- 12897563 TI - The role of nuclear factor-kappa B in interleukin-8 expression by human adenoidal fibroblasts. AB - OBJECTIVES/HYPOTHESIS: The production of cytokines by adenoids is known to be associated with inflammation of nasopharynx and the pathogenesis of otitis media with effusion. However, the role of adenoids in producing inflammatory cytokines such as interleukin-8 (IL-8) is not yet clear. In the present study, expression of IL-8 in adenoidal fibroblasts was investigated at the level of transcription factors. Further, the effects of clarithromycin, a 14-member ring macrolide, on IL-8 gene expression and nuclear factor-kappa B (NF-kappa B) activation in adenoidal fibroblasts were evaluated. STUDY DESIGN: In vitro study for the production of inflammatory cytokine from human adenoidal fibroblasts. METHODS: Adenoidal fibroblasts were incubated with nontypeable Haemophilus influenzae endotoxin or interleukin-1 beta. Then the expression of IL-8 and the influence of NF-kappa B inhibitor and clarithromycin were evaluated. Interleukin-8 protein production was assessed by ELISA, and IL-8 messenger RNA production was measured by Northern blot analysis and reverse transcriptase-polymerase chain reaction. Activation of NF-kappa B and inhibition of its activation were determined by electrophoretic mobility shift assay. RESULTS: The expression of both IL-8 protein and messenger RNA in adenoidal fibroblasts was enhanced by Haemophilus influenzae endotoxin and interleukin-1 beta and was positively correlated with increases in NF-kappa B activity. Treatment of cells with the NF-kappa B inhibitor N-tosyl-(L)-phenylalanine chloromethyl ketone, as well as with clarithromycin, reduced expression of IL-8 and NF-kappa B activity in a dose dependent manner. CONCLUSIONS: Results suggest that adenoidal fibroblasts produce IL-8 in response to endotoxin through NF-kappa B activation. The inhibitory effects of clarithromycin on NF-kappa B activation and IL-8 production in adenoidal fibroblasts might explain, in part, the mechanism of this drug in improving otitis media with effusion. PMID- 12897564 TI - Fiberoptic endoscopic evaluation of swallowing. AB - OBJECTIVES/HYPOTHESIS: Approximately 8 million people in the United States report some degree of dysphagia annually; as the population continues to age, this will become a greater problem. A comprehensibly performed and interpreted fiberoptic endoscopic evaluation of swallowing provides invaluable information for the evaluation and management of individuals with dysphagia or aspiration, or both. STUDY DESIGN: Review. METHODS: The state-of-the-art technique, interpretation, predictive value, and safety of fiberoptic endoscopic evaluation of swallowing was reviewed. PMID- 12897565 TI - Bacteria and granulation tissue associated with Montgomery T-tubes. AB - OBJECTIVES: Although complications (infection, development of granulation tissue) of silicone Montgomery T-tubes have been reported, the microbiological consequences and the origin of granulation tissue have not yet been evaluated. STUDY DESIGN: A prospective trial. METHODS: Twenty-three Montgomery T-tubes from 10 patients were analyzed with regard to the development of granulation tissue, bacterial growth (including genotyping with polymerase chain reaction), and results of sensitivity testing. Furthermore, stent sterilization (n = 6) was investigated. RESULTS: Granulation tissue occurred with 74% of the stents, and all specimens showed signs of infection but no foreign body reaction. The predominant organisms were Staphylococcus aureus (35%) and Pseudomonas aeruginosa (17%). The differences between groups with and without granulation tissue were significant for P aeruginosa. Polymerase chain reaction fingerprinting of the S aureus obtained from 15 stents (n = 3 patients) revealed a total of seven different genotypes. Whereas two of these patients harbored six different genotypes of S aureus, the third patient was persistently colonized by S aureus over a 15-month period with the identical genotype. Susceptibility testing showed most commonly (65%) sensitivity to a combination of amoxicillin-clavulanate and ofloxacin. After sterilization, 92% of analyzed stent segments showed no bacterial growth. CONCLUSIONS: Granulation tissue commonly occurred next to the silicone (subglottic area, stoma) where S aureus and P aeruginosa were commonly isolated. A combination of mechanical irritation and bacterial infection seems to account for the development of granulation tissue. Polymerase chain reaction fingerprinting showed both prolonged persistence and a change of colonizing strains after multiple stent replacements. A combination of amoxicillin clavulanate and ofloxacin is the most effective antibiotic therapy. Sterilization of the cost-intensive silicone stents is feasible, and reuse in the same patient is justifiable from economic aspects. PMID- 12897566 TI - Spatial distribution of tumor vaccine improves efficacy. AB - OBJECTIVES: Genetically engineered tumor cells were used as a vaccine in a murine model to compare tumor formation after inoculating multiple sites versus a single site. The effect of vaccinating draining lymph node basins was evaluated. STUDY DESIGN: Mice were vaccinated in either the floor of the mouth, the draining nodes of the front legs, the hind leg, or a combination of sites. Seven days later, the mice were challenged with parental tumor cells in the floor of the mouth and followed for tumor growth. METHODS: A retroviral vector was used to transduce the granulocyte-macrophage colony-stimulating factor (GM-CSF) gene into SCCFVII/SF tumor cells, which were then irradiated to prevent replication in vivo. Syngeneic C3H/HeJ mice were vaccinated with 1 x 10(6) cells in various sites, then challenged with 1 x 10(5) parental cells after 7 days. RESULTS: Animals vaccinated in multiple sites had better protection from later tumor challenge than those receiving single vaccinations. Of the animals receiving vaccination at multiple sites, those vaccinated in the site of tumor challenge (floor of the mouth) had more protection than those not vaccinated at the site. CONCLUSIONS: Mice vaccinated at multiple draining lymph node sites were better primed against tumor challenge than mice receiving single inoculations. Vaccination strategies that included the challenge site (floor of the mouth) and the nodes near this site were optimal. PMID- 12897567 TI - Expression of extracellular matrix metalloprotease inducer in laryngeal squamous cell carcinoma. AB - OBJECTIVES/HYPOTHESIS: Head and neck cancer tumor cell invasion is responsible for both local destruction and distant metastasis. Invasion is largely mediated by matrix metalloproteases that are thought to be induced by tumor cell derived extracellular matrix metalloprotease inducer (EMMPRIN) in surrounding fibroblasts. We hypothesize that EMMPRIN is overexpressed in laryngeal cancer. STUDY DESIGN: Retrospective analysis of patients with supraglottic laryngeal cancer. METHODS: Total protein immunoblotting and immunohistochemical analysis of normal and malignant tissue were performed to determine EMMPRIN expression. EMMPRIN immunoreactivity in 33 patients was correlated with clinicopathological features and survival. RESULTS: Whole-tissue lysates of tumors (n = 8) and metastatic lymph nodes (n = 2), but not normal skin (n = 8) or mucosa (n = 6), expressed significant amounts of EMMPRIN by immunoblotting. EMMPRIN membrane immunoreactivity (transmembrane EMMPRIN score) was associated with nodal positivity (P =.07), and it was a borderline significant predictor of survival (Hazards Ratio = 2.4; 95% CI, 0.88-6.55). As a categorical variable, higher transmembrane EMMPRIN score was associated with higher mortality. CONCLUSIONS: The present study helps to establish EMMPRIN as a widely expressed protein in dysplastic mucosa and supraglottic laryngeal cancer, but not in normal epithelial counterparts. PMID- 12897568 TI - Parotid carcinoma: impact of clinical factors on prognosis in a histologically revised series. AB - OBJECTIVE: To analyze clinical data and possible prognostic factors of patients with primary carcinoma of the parotid gland. STUDY DESIGN: A retrospective study was made of 85 patients with suspected parotid cancer who were admitted to the Center of Head and Neck Oncology at Odense University Hospital (Odense, Denmark) from 1975 to 1994. METHODS: Patient records were retrospectively reviewed and tabulated for relevant clinical parameters. A complete revision of histological examination was performed by the same pathologist and the tumors were classified according to the 1991 international guidelines from the World Health Organization. Five tumors (6%) did not fulfill the criteria of malignancy and were reclassified as benign. In another five cases the assumed primary parotid carcinomas were found to be metastatic disease from cancers of the breast, prostate, skin, and lungs. Ten patients (12%) were excluded from the analysis, leaving 75 individuals for the study. RESULTS: Twenty-four percent of patients were classified as T1, 32% as T2, 15% as T3, and 28% as T4. One patient (1%) was not classifiable (TX). Regional lymph node metastases were found in 17 cases (23%). Two of these were classified as N1 (3%), 2 as N2A (3%) and 13 as N2B (17%). Four patients were registered as not classifiable (NX) (5%). No patients were classified as N3. Distant metastases were found in four patients (5%) (one patient had bone and liver metastases and three patients had lung metastases). Five-year recurrence-free survival of the entire study group was 63%, disease specific survival was 69%, and crude survival was 52%. In univariate analysis, tumor size, histological appearance, T status, stage, the presence of lymph node metastases, distant metastases, pain, and facial nerve dysfunction had a significant influence on survival. CONCLUSIONS: A thorough histological revision is pivotal in retrospective parotid carcinoma studies, and tumor size; histological appearance; T, N, and M status; stage; facial nerve dysfunction; and pain from the face and/or neck seem to be significant prognostic indicators for patients with primary parotid carcinoma. PMID- 12897569 TI - A new stent for the management of adult and pediatric laryngotracheal stenosis. PMID- 12897570 TI - Sphenopalatine ganglion block before removal of nasal packing. PMID- 12897571 TI - Inferior turbinate flap for repair of nasal septal perforation. PMID- 12897572 TI - The use of "race" for classification in medicine: is it valid? PMID- 12897573 TI - Effects of 0.005% latanoprost on ocular anterior structures and ciliary body thickness. AB - PURPOSE: To assess the effects of 0.005% latanoprost on the anterior segment geometry and ciliary body thickness using ultrasound biomicroscopy. Intraocular pressure, refraction, visual acuity, and pupil size were also evaluated. PATIENTS AND METHODS: Thirty patients with untreated ocular hypertension or primary open angle glaucoma (mean age: 59.3 +/- 9.9 years) were recruited into this prospective, controlled, open trial. Before and after 1 week of 0.005% latanoprost administration, the following parameters were tested: refraction, visual acuity, pupil diameter, intraocular pressure, 5 conventional ultrasonographic A-scan variables, 16 ultrasound biomicroscopy parameters, and the ultrasound biomicroscopy ciliary body thicknesses at a distance of 1500 microns (CBT1), 2000 microns (CBT2), and 2500 microns (CBT3) from the scleral spur. RESULTS: Latanoprost 0.005% caused a marked intraocular pressure-lowering effect in all patients (from 22.8 +/- 3.1 mm Hg to 14.1 +/- 2.9 mmHg; -38%, P < 0.0001), without any refractive, visual acuity, or pupillary alterations. The A scan echobiometry variables were unchanged, while ultrasound biomicroscopy confirmed a significant posttreatment increase of CBT2 (from 434 +/- 140 microns to 536 +/- 127 microns; +102 microns, P = 0.01) and CBT3 (from 319 +/- 103 microns to 412 +/- 100 microns; +93 microns, P = 0.003) compared with controls (CBT2: from 493 +/- 165 microns to 473 +/- 135 microns, -20 microns, P = NS; CBT3: from 388 +/- 130 microns to 365 +/- 87 microns, -23 microns, P = NS). None of the changes observed in the other UBM parameters was statistically significant. No significant correlation was detected between ciliary body thickness increase and intraocular pressure-lowering effect. CONCLUSION: The increase of ciliary body thickness, which was measured in vivo by ultrasound biomicroscopy and associated with the intraocular pressure-lowering effect, indirectly supports the mechanism of uveoscleral outflow enhancement induced by latanoprost. These data are in agreement with the biochemical hypothesis of the passage of the aqueous flow through the extracellular spaces of the ciliary muscle. PMID- 12897574 TI - Estimating normal optic nerve axon numbers in non-human primate eyes. AB - PURPOSE: The goal of the present study is to develop a semi-automated method to estimate accurately, with minimum variance, the total number of axons by counting a subset of the axons within a primate optic nerve. METHODS: Using an imaging analysis system, axons in 50% of the area of cross-sections of the retrobulbar optic nerve from five adult Rhesus monkeys were counted and extrapolated as an estimate of total axon number of the optic nerves. Both neural and non-neural areas were sampled. With the coordinates of the counts topographically registered, axon numbers within areas ranging from 1 to 50% were resampled. A Monte Carlo and theoretical estimate of the standard deviation of the total axon count for each sampled area was computed. RESULTS: The mean cross-sectional area of the five optic nerves counted was 7.26 +/- 0.6 mm2, and the mean total axon count of the optic nerve area was 1,304,8168 +/- 89,112. When sampling less than 8% of the optic nerve, the standard deviation within the individual of the total estimated axon number increased sharply. CONCLUSION: With this technique, the variance within each individual increased only slightly when the counting area was reduced from 50 to 8%, but increased sharply when the counted area became less than 8%. While counting less than 8% of the optic nerve area gives a good estimation of total axon count, the effect of a substantial increase in the standard deviation on the statistical power needed to differentiate group differences will depend on the study design. PMID- 12897575 TI - Long-term effect of retinal ganglion cell axotomy on the histomorphometry of other cells in the porcine retina. AB - PURPOSE: To determine the effect of retinal ganglion cell axotomy on the thickness of inner plexiform, inner nuclear, and outer plexiform layers, as well as the densities of short- and middle-to-long-wavelength cones, in the porcine retina. METHODS: Unilateral retinal ganglion cell axotomy was performed in seven domestic pigs by either surgical optic nerve section or peripapillary argon laser photocoagulation. Damage to the retinal vasculature was ruled out with fluorescein angiography. Histologic examination of the retinal tissue was performed nine months later. Cone densities were determined immunohistochemically with the anti-visual pigment antibodies COS-1 and OS-2. Image analysis of semithin retinal cross sections was used to measure the thickness of the retinal layers. The effect of axotomy was quantified by optic nerve axon counts and estimations of retinal ganglion cell counts. The data were compared between the eyes with axotomy and the contralateral normal eye using the nonparametric Wilcoxon rank sum test. RESULTS: Treatment of the peripapillary retina with the argon laser resulted in a median decrease in axon counts and retinal ganglion cell density estimates of 31%. No optic nerve axons and cells resembling retinal ganglion cells were found in the eyes with transected optic nerves. There was no significant difference in either the thickness of any retinal layers or cone densities between axotomized and normal control eyes. CONCLUSION: No signs of retrograde transsynaptic degeneration were observed in porcine retinas nine months after retinal ganglion cell axotomy. PMID- 12897576 TI - Laser suture lysis after trabeculectomy. AB - PURPOSE: The author attempted to avoid ocular hypotony following a mitomycin-C trabeculectomy by controlling the interval between surgery and laser suture lysis. METHODS: When 10 seconds of digital pressure on the eye following a mitomycin-C trabeculectomy produced a 20% drop of the intraocular pressure, the decision of whether or not to perform laser suture lysis was delayed until the following examination. RESULTS: Twenty-nine consecutive eyes (27 patients) with primary open-angle glaucoma underwent mitomycin-C trabeculectomy in which laser suture lysis was required during the postoperative period to reach the target intraocular pressure. The eyes were classified by the severity of the glaucoma: mild, moderate, and severe. For moderate and severe glaucoma, an unqualified success was defined as an intraocular pressure of 10 +/- 2 mm Hg. A qualified success had an intraocular pressure of 5 to 7 mm Hg or 13 to 15 mm Hg. Failure was defined as an intraocular pressure less than 5 mm Hg or greater than 15 mm Hg. Fifteen eyes were called success, 7 eyes qualified success, and 7 eyes failure. When the final laser suture lysis was carried out within the first two postoperative months, two of the failure eyes sustained hypotony. If the final laser suture lysis had been carried out after the first two postoperative months, there were no eyes with hypotony. There was no statistical difference in the final intraocular pressure between the early laser suture lysis and late laser suture lysis groups. CONCLUSIONS: From this pilot study it would appear that delaying the final laser suture lysis following trabeculectomy (MMC) until after the second postoperative month may reduce the risk of hypotony without adversely affecting the final intraocular pressure. PMID- 12897577 TI - Which endpoint: mean intraocular pressure or proportion of successful patients? AB - A reduction in intraocular pressure in clinical trials can be determined through the mean intraocular pressure, through the proportion of patients who have the intraocular pressure reduced to a specific target intraocular pressure, or both. Since both these possible endpoints measure the shift of 2 intraocular pressure distributions, we recommend that only one of them be tested. In general, testing the difference between mean-values is much more efficient than testing the difference between proportions. However, proportions of successful patients are valuable in showing the clinical implication of a reduction in mean intraocular pressure, particularly when evaluating a moderate pressure reduction. The effect of a small mean intraocular pressure reduction on the probability to reach the target intraocular pressure is pointed out, particularly the fact that it can be substantial even if the mean reduction is smaller than the measurement error. PMID- 12897578 TI - Comparison between phacoemulsification and the Blumenthal technique of manual small-incision cataract surgery combined with trabeculectomy. AB - OBJECTIVE: To compare results of filtration combined with either phacoemulsification or the Blumenthal technique of manual small-incision cataract surgery. MATERIALS AND METHODS: Records of glaucoma "triple" operations from March 1997 to May 2000 were reviewed. Seventy-eight eyes (70 patients) that underwent phaco-triple were compared with 86 eyes (80 patients) that underwent the Blumenthal technique of manual small-incision cataract surgery combined with filtration (Blumenthal triple). Three minutes of 0.4 mgs/ml Mitomycin was used in all eyes. Posterior chamber IOLs were implanted through 5.5-mm incisions in both groups. Outcome measures were intraocular pressure (IOP) reduction and achievement of target IOP. Fourteen patients who underwent phacoemulsification triple in one eye and Blumenthal triple in the other eye were also evaluated separately. RESULTS: The minimum follow-up period was 6 months (range 6-30 months). At last follow-up review, mean reduction in IOP was 17.7 mm Hg (+/- 9.3 mm Hg) in the phaco group and 17.1 mm Hg (+/- 10 mm Hg) in the Blumenthal group. At last visit, target IOP was achieved in 75.6% of the phaco group and 73% of the Blumenthal group. There was no significant difference between groups in IOP reduction or achievement of target IOP. In the 14 patients who had undergone phaco-triple in one eye and Blumenthal-triple in the other, there was no inter eye difference in IOP reduction. CONCLUSIONS: In this small retrospective study we could not demonstrate a difference in IOP outcomes between the two procedures. PMID- 12897579 TI - New glaucoma classification method based on standard Heidelberg Retina Tomograph parameters by bagging classification trees. AB - PURPOSE: In this article we propose and evaluate nonparametric tree classifiers that can handle non-normal data and a large number of possible predictors using the full set of standard Heidelberg Retina Tomograph measurements for classifying glaucoma. METHODS: The classifiers were trained and tested using standard Heidelberg Retina Tomograph parameters from examinations of 98 subjects with glaucoma and 98 normal subjects of the Erlangen Glaucoma Registry. All patients and control subjects were evaluated by 15 degrees -optic disc stereographs, Heidelberg Retina Tomograph measurements, standard computerized white-in-white perimetry, and 24-hour-intraocular pressure profiles. The subjects were matched by age and sex. Standard classification trees as well as bagged classification trees were used. The classification outcome of the trees was compared with the classification by two published linear discriminant functions based on Heidelberg Retina Tomograph variables with respect to their cross-validated misclassification error. RESULTS: The bagged classification tree had the lowest misclassification error estimate of 14.8% with a sensitivity of 81.6% at a specificity of 88.8%. The cross-validated error rates of the two linear discriminant function procedures were 20.4% (sensitivity 82.6%, specificity 76.7%) and 20.6% (sensitivity 81.4%, specificity 77.3%) for our set of observations. Bagged classification trees were able to reduce the misclassification error of glaucoma classification. CONCLUSIONS: Bagged classification trees promise to be a new and efficient approach for glaucoma classification using morphometric 2- and 3-dimensional data derived from the Heidelberg Retina Tomograph, taking into account all given variables. PMID- 12897580 TI - Replacing maximum-tolerated medications with latanoprost versus adding latanoprost to maximum-tolerated medications: a two-center randomized prospective trial. AB - PURPOSE: To compare the replacement of every drug with latanoprost 0.005% once a day in glaucomatous eyes with poorly controlled intraocular pressure upon combination therapy, versus addition of latanoprost to the pre-existing treatment. PATIENTS AND METHODS: STUDY DESIGN: prospective, investigator-masked, two-center, randomized clinical trial lasting 3 months. ELIGIBILITY CRITERIA: open-angle glaucoma; IOP > or = 21 mm Hg upon the combination of a non-selective beta-blocker with pilocarpine or dorzolamide or both; no previous bulbar surgery; and prior glaucoma therapy lasting at least 2 years. Two treatment arms: (1) addition of latanoprost 0.005% QD to the pre-existing therapy [group A]; (2) substitution with latanoprost alone [group B]. RESULTS: One hundred thirty-six eyes (68 eyes/treatment group) were randomized according to intraocular pressure level and the number of adjunctive medications to beta blocker. Both treatments provided a significant IOP decrease over baseline (from 23.5 +/- 1.4 to 19.7 +/- 1.9 mm Hg in group A, (P < 0.001); from 23.2 +/- 1.3 to 20.1 +/- 2.2 mm Hg in group B (P < 0.001), paired Student t test). At the end of the follow-up period, group A showed a higher number of intraocular readings less than or equal to 18 mm Hg than group B (42.6% vs. 30.8%; Fisher exact test: P = 0.018). CONCLUSIONS: In eyes showing an intraocular pressure greater than 21 mm Hg upon combination therapy, the substitution of the pre-existing treatment with latanoprost can provide a significant IOP decrease. However, adding latanoprost to the pre existing therapy is more likely to achieve a target intraocular pressure less than or equal to 18 mm Hg. PMID- 12897581 TI - Suprachoroidal seton implantation in refractory glaucoma: a novel surgical technique. AB - PURPOSE: To describe a new surgical technique in which aqueous humor is diverted from the anterior chamber to the suprachoroidal space for the augmentation of uveoscleral outflow in the management of refractory glaucoma. METHODS: Four painful-blind eyes of four consecutive patients were included in the study. Mean age of patients was 54.7+/- 9.2 years. Preoperative diagnosis was neovascular glaucoma complicating diabetic retinopathy in three cases and chronic angle closure glaucoma in one case. Mean preoperative intraocular pressure of the patients receiving two medications was 58.5 +/- 9.2 mm Hg. A modified Krupin eye valve with disk was implanted into the suprachoroidal space. The anterior tube part of the Seton device was placed into the anterior chamber through the long scleral tunnel for draining the aqueous humor from the anterior chamber to the suprachoroidal space. RESULTS: The placement of modified Krupin eye valve with disk to the suprachoroidal space was achieved in all cases. While mean preoperative intraocular pressure was 58.5 +/- 9.2 mm Hg, it was 14.2 +/- 4.7 mm Hg at postoperative one week. It was 13.5 +/- 4.6 mm Hg and 15 +/- 4.9 mm Hg at one and three months respectively. At the last follow-up visit, mean intraocular pressure was 17.25 +/- 5.37 mm Hg ranging from 12 to 24 mm Hg. Choroidal detachment was developed in one case and regressed in six weeks. Rubeosis irides regressed at third month in three cases. None of the eyes developed suprachoroidal hemorrhage, retinal detachment, or phthisis bulbi. CONCLUSION: The drainage of aqueous humor from the anterior chamber to the suprachoroidal space with the implantation of the glaucoma Seton device is effective in lowering intraocular pressure in refractory glaucoma. PMID- 12897582 TI - Diode laser transscleral cyclophotocoagulation in the treatment of chronic angle closure glaucoma: a preliminary study. AB - PURPOSE: To evaluate the efficacy and safety of diode laser transscleral cyclophotocoagulation in the treatment of chronic angle-closure glaucoma. PATIENTS AND METHODS: A prospective, non-comparative interventional pilot study was conducted. Fourteen eyes of 14 Chinese patients with chronic angle-closure glaucoma whose intraocular pressures were greater than 21 mm Hg on medications were treated with diode laser transscleral cyclophotocoagulation using the G probe at the United Christian Hospital between February 2000 and May 2001. The inferior 270 degrees quadrant (from 2 to 11 o'clock for right eye and from 1 to 10 o'clock for left eye) was treated and the patients were followed up regularly. The initial laser energy was set at 2,000 mWatt with a duration of 2 seconds. The post-treatment anti-glaucoma medications were adjusted according to the intraocular pressure. If medications failed to lower the intraocular pressure to below 21 mm Hg, cyclophotocoagulation to the same inferior 270 degrees quadrant was repeated. RESULTS: All patients completed a 12-month follow-up period. The total success rate defined as IOP < 21 mm Hg with or without medication(s) was 85.7% at 1 year of follow-up review. The mean +/- SD intraocular pressure decreased from pre-treatment level of 36.9 +/- 11.7 mm Hg to 18.9 +/- 6.5 mm Hg at 1 year after treatment. The difference was statistically significant (P < 0.001) (Paired t test). The mean +/- SD number of IOP-lowering eyedrops was significantly reduced from 1.9 +/- 0.7 before cyclophotocoagulation to 0.4 +/- 0.8 at 1 year after treatment (P = 0.0002) (Paired t test). Two eyes required repeat treatment. Seven eyes (50%) had atonic pupil following the laser treatment. CONCLUSION: Diode laser transscleral cyclophotocoagulation is effective in lowering the intraocular pressure in chronic angle-closure glaucoma and its effect lasts for at least 1 year. PMID- 12897583 TI - Intraocular pressure in a Somali population living in the United States. AB - PURPOSE: Intraocular pressure is a risk factor for the development of glaucomatous optic neuropathy. With few exceptions, higher mean intraocular pressure and greater prevalences of glaucoma have been reported for individuals of African origin. This study was performed to compare the mean intraocular pressure of a group of ethnic East Africans living in the United States with that of Caucasians living in the same community. PATIENTS AND METHODS: Retrospective comparison of Somali patients 30 years of age or greater with age- and gender matched Caucasian control subjects. All patients were seen in the same clinic in Seattle, Washington, between July 1996 and March 1998. Patients were excluded for conditions or medications affecting intraocular pressure and for a diagnosis of glaucoma. Mean intraocular pressure of the two populations was compared using an independent sample two-tailed t test. RESULTS: Following exclusions and defined age limits, 57 Somali patients and 57 Caucasian control subjects were included in the final analysis. Mean age of Somalis was 48.5 +/- 12.2 years versus 48.1 +/- 11.3 years for control subjects (P = 0.84). The mean intraocular pressure of Somalis was 13.76 +/- 3.63 mm Hg versus 13.94 +/- 2.78 mm Hg for control subjects (P = 0.77). CONCLUSION: Some populations of African origin may not have higher mean intraocular pressures compared with non-African populations. Consideration of ethnic and racial origins more specific than "African" should be given when evaluating intraocular pressure in individual patients. PMID- 12897584 TI - Episcleral venous pressure in younger and older subjects in the sitting and supine positions. AB - PURPOSE: Intraocular pressure is higher in older than younger subjects during the day and night. We attempted to determine whether episcleral venous pressure could explain the difference in the sitting and supine positions. PARTICIPANTS AND METHODS: We compared episcleral venous pressure, intraocular pressure, and blood pressure in a group of younger subjects aged 18 to 30 years and in a group of older subjects aged 65 years or older. The above measurements were performed in the sitting position and after 15 minutes in the supine position. RESULTS: The main difference between the groups was their systemic conditions. Comparing both groups, episcleral venous pressure and intraocular pressure were not different in the sitting position. However, in the supine position, episcleral venous pressure (9.6 vs. 8.3 mm Hg) (P < 0.01) and intraocular pressure (17.1 vs. 15.6 mm Hg) (P < 0.05) were higher in the older group. Statistically, a within-group analysis showed a significant increase in intraocular pressure in the supine position for both the younger (+ 0.8 mm Hg) (P < 0.001) and older subjects (+ 1.8 mm Hg) (P < 0.02). This was associated with an increased episcleral venous pressure in younger (+ 0.4 mm Hg) (P < 0.001) and older subjects (+1 mm Hg) (P < 0.02). There was no gender difference in intraocular pressure and episcleral venous pressure. No differences were found for intraocular pressure and episcleral venous pressure in subjects having certain systemic conditions. Blood pressure was higher for older subjects (P < 0.001). It decreased in the supine position for both groups (P < 0.001). CONCLUSION: Intraocular pressure and episcleral venous pressure were not different in the younger and older group in the sitting position. They were higher in the supine position for older subjects. There was no gender difference. PMID- 12897585 TI - Hypotony maculopathy and papilledema after trabeculectomy in a patient with pseudotumor cerebri. PMID- 12897586 TI - Relationship between postural change of the intraocular pressure and visual field loss in primary open-angle glaucoma. AB - PURPOSE: To determine the relationship between the postural changes of the intraocular pressure and the visual field loss in patients with primary open angle glaucoma. METHODS: Eleven normal subjects and 11 patients with primary open angle glaucoma were studied. Intraocular pressure was measured in both the sitting and the supine positions. Visual fields were measured with automated perimetry. RESULTS: When patients moved from a sitting to supine position, the intraocular pressure increased by an average of 3.1 +/- 0.4 mm Hg (mean +/- SEM) in normal subjects and 4.0 +/- 0.2 mm Hg in patients with primary open-angle glaucoma. There was a significant difference between the normal subjects and patients with primary open-angle glaucoma (P = 0.049). Intraocular pressure increased by 4.4 +/- 0.3 mm Hg (P = 0.02) in the worse eye for mean deviation and 3.6 +/- 0.3 mm Hg (P = 0.38) in the better eye for mean deviation. There was no significant difference in intraocular pressure in the sitting position between both groups. CONCLUSIONS: The greatest difference in intraocular pressure between the sitting and supine positions was observed in the worse eye of patients with primary open-angle glaucoma. This result suggests that damage to the optic nerve in primary open-angle glaucoma might occur when patients are asleep in the supine position. PMID- 12897587 TI - Unoprostone isopropyl ester darkens iris color in pigmented rabbits with sympathetic denervation. AB - PURPOSE: Unoprostone isopropyl ester (unoprostone) -induced iris color darkening was evaluated in a rabbit model using a cyclooxygenase inhibitor, an alpha(1) adrenergic antagonist, and sympathetic denervation. MATERIALS AND METHODS: Dutch belted rabbits were divided into five groups based on type of surgery and eyedrop treatment to both eyes: (1) sham surgery (n = 7); (2) bilateral superior cervical ganglionectomy (SCGx, n = 7); (3) SCGx plus flurbiprofen 0.03% (n = 7); (4) SCGx plus thymoxamine 0.5% (n = 6); and (5) SCGx plus flurbiprofen and thymoxamine (n = 6). All rabbits were treated with unoprostone 0.12% to one eye and its vehicle to the contralateral eye twice daily for 43 weeks after SCGx. Periodic color photographs of paired eyes were scored for difference in eye color. Iris melanin and aqueous humor protein were measured at week 43. RESULTS: Twenty-three of the 26 rabbits with bilateral SCGx and unilateral unoprostone treatment demonstrated a darker iris color on the unoprostone-treated side. The average scores (demonstrating difference in iris color) comparing photographs of treated versus control eyes in the four SCGx groups were higher than those in the sham surgery group (P < 0.03), and higher than at week 0 (P < 0.001). The group pretreated with flurbiprofen and thymoxamine had the highest score of all groups. The aqueous humor protein in unoprostone-treated eyes was higher (P < or = 0.0001) than in vehicle-treated eyes. The melanin content of irides of the denervated groups was higher (P < or = 0.01) in unoprostone-treated than in vehicle-treated eyes. CONCLUSION: Unoprostone produced iris color darkening in pigmented rabbit eyes with sympathetic denervation. Pretreatment with flurbiprofen and thymoxamine appeared to enhance this effect but this was not statistically demonstrated by the study. PMID- 12897588 TI - Laser suture lysis for non-valved aqueous drainage implants. AB - PURPOSE: To describe a technique for laser suture lysis with non-valved aqueous drainage implants. METHODS: The surgical juxtaposition of the aqueous drainage tube and notched scleral graft allows the ligature to be placed in a predictable position aiding laser suture lysis. RESULTS: After drainage device encapsulation limits the possibility of hypotony, this technique allows easy postoperative lysis of drainage tube ligatures. CONCLUSION: This simple modification of surgical technique aids surgeons in finding and releasing aqueous drainage device ligature sutures. PMID- 12897589 TI - Writing for Clinical Orthopaedics and Related Research. PMID- 12897590 TI - Evidence-based orthopaedics: a paradigm shift. PMID- 12897591 TI - The history of instrumental precision in medicine. Univ Med Magazine, October 1841. PMID- 12897592 TI - Hierarchy of evidence: from case reports to randomized controlled trials. AB - In the hierarchy of research designs, the results of randomized controlled trials are considered the highest level of evidence. Randomization is the only method for controlling for known and unknown prognostic factors between two comparison groups. Lack of randomization predisposes a study to potentially important imbalances in baseline characteristics between two study groups. There is a hierarchy of evidence, with randomized controlled trials at the top, controlled observational studies in the middle, and uncontrolled studies and opinion at the bottom. This hierarchy has not been supported in two recent publications in the New England Journal of Medicine which identified nonsignificant differences in results between randomized, controlled trials, and observational studies. The current authors provide an approach to organizing published research on the basis of study design, a hierarchy of evidence, a set of principles and tools that help clinicians distinguish ignorance of evidence from real scientific uncertainty, distinguish evidence from unsubstantiated opinions, and ultimately provide better patient care. PMID- 12897593 TI - Methodologic issues in randomized controlled trials of surgical interventions. AB - A physician's ability to make conclusions about the relative efficacy of interventions based on their clinical experience is limited frequently. Therefore, surgeons commonly use research evidence to guide their clinical practice. The randomized controlled trial is the strongest study design. However, randomization in itself does not guarantee the trial results are valid (free from bias). Understanding the potential impact of various methodologic features of a randomized controlled trial allows a clinician to determine the validity of a trial. We present a guide for evaluating the validity of randomized controlled trials giving special consideration to issues confronted in surgical trials. PMID- 12897594 TI - Methodologic issues in observational studies. AB - Assessment of orthopaedic treatments depends primarily on studies that analyze data abstracted from medical records or collected prospectively for research purposes. Because treatment in these studies is determined by the surgeon and the patient rather than controlled by the researcher, these studies are referred to as observational. Most orthopaedics research uses the observational study design because it is less expensive and more timely, includes more representative patients, and leaves medical decision-making under the control of the physician and the patient. Biases associated with observational studies are influenced greatly by study design, methods of data collection, and statistical analysis. All observational studies should report factors influencing relevance (characteristics of patients, treatments, and outcome measures), and factors that influence validity (methods to obtain information and patient risk factors that may differ between treatments). Improved standards for reporting observational studies will facilitate the interpretation of results and the comparison of studies with conflicting results. PMID- 12897595 TI - Methodologic issues in systematic reviews and meta-analyses. AB - Systematic reviews of original research are increasing in number. Systematic reviews are distinct from narrative reviews because they address a specific clinical question, require a comprehensive literature search, use explicit selection criteria to identify relevant studies, assess the methodologic quality of included studies, explore differences among study results, and either qualitatively or quantitatively synthesize study results. Systematic reviews that quantitatively pool results of more than one study are called meta-analyses. Several organizations are collaboratively involved in producing high quality systematic reviews and meta-analyses. Familiarity with how to do a systematic review and meta-analysis will lead to greater skill in using this type of article. For clinicians, teachers, and investigators, systematic reviews and meta analyses are useful sources of evidence. PMID- 12897596 TI - Statistical sampling and hypothesis testing in orthopaedic research. AB - The purpose of the current article was to review the process of hypothesis testing and statistical sampling and empower readers to critically appraise the literature. When the p value of a study lies above the alpha threshold, the results are said to be not statistically significant. It is possible, however, that real differences do exist, but the study was insufficiently powerful to detect them. In that case, the conclusion that two groups are equivalent is wrong. The probability of this mistake, the Type II error, is given by the beta statistic. The complement of beta, or 1-beta, representing the chance of avoiding a Type II error, is termed the statistical power of the study. We previously examined the statistical power and sample size in all of the studies published in 1997 in the American and British volumes of the Journal of Bone and Joint Surgery, and in Clinical Orthopaedics and Related Research. In the journals examined, only 3% of studies had adequate statistical power to detect a small effect size in this sample. In addition, a study examining only randomized control trials in these journals showed that none of 25 randomized control trials had adequate statistical power to detect a small effect size. However, beta, or power, is less well understood. Because of this, researchers and readers should be aware of the need to address issues of statistical power before a study begins and be cautious of studies that conclude that no difference exists between groups. PMID- 12897597 TI - The risk of false-positive results in orthopaedic surgical trials. AB - The risk of concluding that the results of a particular study are true, when, in fact, they really are attributable to chance (or random sampling error) is underappreciated by investigators. This erroneous false-positive conclusion is designated as a Type I or alpha error. The extent to which randomized trials in surgery risk Type I errors is unclear. The current authors hand-searched four orthopaedic journals, six general surgery journals, and five medical journals to identify recently published randomized trials (within the past 2 years). Information on outcomes and statistical adjustment for multiple outcomes was recorded for each study. The risk of a Type I error was calculated for each study that did not explicitly state a primary outcome measure for the main statistical comparison. One hundred fifty-nine studies met the inclusion criteria for the study: 60 studies from orthopaedic journals, 49 studies from nonorthopaedic surgical journals, and 50 studies from medical journals. Of the trials that did not state a primary outcome measure, the risk of Type I errors (false-positive results) in orthopaedic and nonorthopaedic surgery journals (mean 37.3% +/- 13.3% and 37.6% +/- 10.5%, respectively) were significantly greater than medical journals (10.1% +/- 1.9%). In the current review of randomized trials in surgery and medicine, the following is reported: (1) reporting of primary outcomes in trials was inadequate; (2) one in three trials in surgery and one in 10 trials in medicine risked false-positive results; and (3) few trials in surgery and medicine considered adjustment for multiple comparisons. PMID- 12897598 TI - Common statistical methods in orthopaedic clinical studies. AB - We discuss the statistical representation and management of random error in orthopaedic clinical studies. Descriptive studies (such as case series) collect information about a sample that may be generalized to describe a population. Typically this description is in the form of summary statistics, such as means, proportions, or rates. Error in these variables may be represented by confidence intervals. Correlation and regression are techniques for investigation of the relationship between two or more variables. Descriptive statistics, comparisons of groups, especially hypothesis tests, and assessment of association including correlation and regression are important statistical concepts for clinicians involved in the conduct or appraisal of orthopaedic clinical research. PMID- 12897599 TI - A guide to health measurement. AB - Limited healthcare dollars have resulted in insistence that the benefit of new therapies be evaluated before being approved for marketing or reimbursement under health service systems. Adequate evidence of a treatment's effectiveness includes evidence of impact on patient's health-related quality of life, including physical, mental, and emotional health. There are two types of measures of health related quality of life. One, general health and utility measures, inquire about health in a broad sense, and can be applied and compared across many situations. The second type, specific measures, addresses narrower aspects of life related to a specific problem, function, or manifestations of an underlying disease process. Results of studies focusing on health-related quality of life only will be useful if the measurement instrument is valid and capable of detecting important change. Investigators should make a good choice of measurement instrument, and then ensure their study design will yield valid results. We offer basic guidelines for the measurement of health-related quality of life as an outcome in clinical research. This discussion addresses clinicians, who are making decisions regarding the relevance of study results, and investigators who are designing studies. PMID- 12897600 TI - Measures of health-related quality of life and physical function. AB - Outcome measures may be seen as windows, each of which provides a different perspective on a single view, the impact of a disease or disorder on the patient concerned. A comparison of the orthopaedic literature in 1991 with that in 2001 suggests that the intervening decade saw an increasing interest in the use of patient-based measures in clinical outcome studies, particularly randomized controlled trials. The tool most commonly used to determine the patient's point of view was the generic Short Form-36 (SF-36). Other measures included regional assessments such as the Musculoskeletal Functional Assessment, or the Disability of the Arm, Shoulder and Hand. The Western Ontario McMaster Osteoarthritis Index (WOMAC) (hip and knee), the Simple Shoulder Test (shoulder) and the Roland-Morris Questionnaire or Oswestry Disability Index (low back) were the most common joint specific measures. Each of the questionnaires reported was supported by evidence of reliability and validity, and in most cases one or more studies had shown responsiveness in at least musculoskeletal disorders. We provide a brief description of the most common tools, and review the evidence that orthopaedic research is making increased use of measures of health status and function. PMID- 12897601 TI - It is money that matters: decision analysis and cost-effectiveness analysis. AB - Clinical decisions must be made, often under circumstances of uncertainty and limited resources. Decision analysis and cost-effectiveness analysis are methodologic tools that allow for quantitative analysis and the optimization of decision-making. These methods can be useful for decisions regarding individual patient evaluation and treatment options or in formulating healthcare policy. We overview the methodology of expected value decision analysis and of cost effectiveness analysis, including cost-identification, cost-effectiveness, cost benefit, and cost-utility analyses. Examples are provided of these methods and a user's guide to cost-effectiveness analysis is outlined. PMID- 12897602 TI - Evidence-based orthopaedics: a primer. AB - Evidence-based medicine has evolved from the need of solving clinical problems. In contrast to the traditional paradigm of clinical practice, evidence-based medicine acknowledges that intuition, clinical experience, and pathophysiologic rationale are not sufficient for making the best clinical decisions. Although evidence-based medicine recognizes the importance of clinical experience, it includes the evaluation of evidence from clinical research and the integration of patients' values, preferences, and actions for best clinical decision-making. To optimize this process, evidence-based medicine advocates that a formal set of rules must accompany training and clinicians' common sense to interpret and apply evidence from clinical research results effectively. We describe the critical appraisal of studies related to prognosis and therapy or prevention building on an example relevant for the clinical orthopaedist. Based on the example, the authors describe how clinicians can apply measures of association and of intervention effects to their practice and patient care. The authors conclude with describing the appraisal of systematic reviews, their application to the development of practice guidelines, and the process of guideline development and recommendations. PMID- 12897603 TI - Finding current evidence: search strategies and common databases. AB - With more than 100 orthopaedic, sports medicine, or hand surgery journals indexed in MEDLINE, it is no longer possible to keep abreast of developments in orthopaedic surgery by reading a few journals each month. Electronic resources are easier to search and more current than most print sources. We provide a practical approach to finding useful information to guide orthopaedic practice. We focus first on where to find the information by providing details about many useful databases and web links. Sources for identifying guidelines, systematic reviews, and randomized controlled trials are identified. The second section discusses how to find the information, from the first stage of formulating a question and identifying the concepts of interest, through to writing a simple strategy. Sources for additional self-directed learning are provided. PMID- 12897604 TI - Designing, conducting, and evaluating journal clubs in orthopaedic surgery. AB - The first record of a journal club was that founded in 1875 by Sir William Osler at McGill University for the purchase and distribution of periodicals to which he could not afford to subscribe as an individual. Evidence-based medicine is becoming an accepted educational paradigm in medical education at various levels. An analysis of the literature related to journal clubs in residency programs in specialties other than orthopaedic surgery reveals that the three most common goals were to teach critical appraisal skills (67%), to have an impact on clinical practice (59%), and to keep up with the current literature (56%). The implementation of the structured article review checklist has been found to increase resident satisfaction and improves the perceived educational value of the journal club without increasing resident workload or decreasing attendance at the conference. Periodic evaluation of the conference and the institution of appropriate changes ensures that the journal club remains a valuable and successful part of the training program. PMID- 12897605 TI - Glossary of evidence-based orthopaedic terminology. AB - As surgeons incorporate the philosophy of evidence-based orthopaedics into patient care, they need to become familiar with common terminology. A glossary of commonly used terms encountered among evidence-based orthopaedic practitioners is presented. PMID- 12897606 TI - Pectoralis major rupture in elderly patients: a clinical study of 13 patients. AB - The purpose of this study was to analyze and describe circumstances and clinical aspects of pectoralis major rupture in elderly patients compared with younger patients. Eleven patients were nursing home residents and two were community residents. All had severe pain with a large pectoral ecchymosis extending to the lateral chest wall and arm. Twelve patients experienced substantial blood loss (average, 2.3 g/dL hemoglobin), which resulted in hemodynamic decompensation in two of the patients, necessitating a blood transfusion. The most probable mechanism of injury was indirect, caused by a brisk tearing movement applied to stiff, atrophic muscle during commonly used nursing procedures for transferring, positioning, and dressing the patients. The authors suspect this injury to be more common than reported. Although surgical repair is not required in the elderly, a proper diagnosis is imperative because the injury has a severe clinical impact necessitating care and followup. PMID- 12897607 TI - Radial nerve transposition with humeral fracture fixation: preliminary results. AB - Humeral shaft fractures traditionally have been managed with closed treatment. In patients with polytrauma, open fractures, and patients at risk for nonunion, open reduction and internal fixation and intramedullary nailing have been advocated. The current study describes a technique used in humeral shaft fractures that reduces the risk of iatrogenic radial nerve injury during plate osteosynthesis in fracture patterns at high risk of nonunion (highly comminuted, transverse fractures). Ten patients who had radial nerve transposition were reviewed retrospectively using the electronic records database at the authors' institution. The average age of the patients was 27 years and average followup was 25 months. All had humeral shaft fractures, AO class A3.2 in four patients, B3.2 in five patients, and C2.2 in one patient. All fractures were deemed to be at high risk for nonunion. There were no iatrogenic nerve palsies as a result of the transposition, and no infections. Two patients had delayed or nonunion, who achieved healing after a second intervention. Transposition of the radial nerve is a useful adjunct to plating of humeral shaft fractures in patients at high risk for nonunion. The technique is safe, does not cause iatrogenic injury, and protects the radial nerve during all subsequent approaches to the fracture site. PMID- 12897608 TI - Revision total hip arthroplasty: hospital cost and reimbursement analysis. AB - Revision total hip arthroplasty is a complex surgical procedure that frequently requires high levels of hospital resources. The purpose of the current study was to report the actual costs and reimbursement to the hospital for a stratified group of patients having revision total hip arthroplasty based on a severity index. The clinical and financial records of 49 patients (51 hips) stratified by complexity of revision were reviewed. Clinical variable included age, length of stay, operating time, estimated blood loss, number of transfusions, implant type, metallic augmentation, use of bone graft, and time spent in the postanesthesia care unit. Financial review included the actual fixed and variable costs associated with each procedure. Hospital costs associated with revision total hip arthroplasty were significantly greater in the most complex revisions and in older patients. The use of bone grafting techniques on the femur resulted in significantly greater costs. The average loss to the hospital was $5402 US dollars per procedure with a range of $5657 (US dollars) profit to $28,780 (US dollars) loss. Procedures in patients younger than 65 years has an average loss of $1133 US dollars. All procedures in patients who were 65 years or older resulted in a loss to the hospital, with the average loss being $8617 US dollars. Despite improvements in length of stay, use of clinical pathways, and negotiated discounts on implants, the hospital loss on each hip revision procedure averaged $5402 US dollars. PMID- 12897609 TI - Human leukocyte antigen and aseptic loosening in Charnley total hip arthroplasty. AB - Human leukocyte antigen phenotypes and radiographic course were studied in 70 Japanese patients (90 hips) who had Charnley low friction arthroplasty. Of the 28 hips diagnosed as radiographically loose less than 15 years after surgery, seven hips (25.0%) were in patients who had a positive human leukocyte antigen A31(19). Of the 62 hips that were radiographically stable more than 15 years, two hips (3.2%) were in patients who had a positive human leukocyte antigen A31(19). Of the nine hips in patients who were positive to human leukocyte antigen A31(19), seven hips were diagnosed as radiographically loose (loosening rate was 77.8%) 15 years after surgery. On the other hand, of the 81 hips in the patients who were negative to human leukocyte antigen A31(19), only 21 hips were diagnosed as radiographically loose (loosening rate was 25.9%) 15 years after surgery. These data suggest that individual immune response to implant materials, linked to human leukocyte antigen, may participate in the process of aseptic loosening in Charnley low friction arthroplasty. PMID- 12897610 TI - Improvement in driving reaction time after total hip arthroplasty. AB - Decisions made regarding resumption of driving after total hip arthroplasty may be determined by a combination of factors including driving reaction time and when postsurgical precautions need no longer be adhered to. Ninety patients, ranging in age from 34 to 85 years old were recruited after total hip arthroplasty to measure driving reaction time preoperatively and from 1 to 52 weeks postoperatively. Driving reaction time worsened 1 week postoperatively for patients who had a right hip arthroplasty. The driving reaction time then improved up to 1 year postoperatively. Patients who had a left arthroplasty improved from 1 week postoperative. In general, patients reach their preoperative driving reaction time 4 to 6 weeks postoperatively and continue to improve. PMID- 12897611 TI - Acetabular rim degeneration: a constant finding in the aged hip. AB - During recent years the acetabular labrum has gained increased interest because its degeneration frequently is found in association with early osteoarthritis of the hip. To determine spatial distribution of labral degeneration in the aged hip and to identify the pathologic features triggering this event, an anatomic postmortem and an intraoperative in vivo study were done in 30 cadavers (range, 60-90 years) and in 18 elderly patients (range, 69-97 years) who had hemiarthroplasty for displaced femoral neck fractures. In both groups, no gross anatomic or radiographic abnormalities suggesting advanced osteoarthritis were present. All cadaveric acetabuli (30 of 30) revealed labrum and cartilage damage. Labrum damage (17 of 18) and cartilage lesions (16 of 18) occurred less frequently and were smaller in the elderly patients. Peripheral joint degeneration was most frequent at the superior acetabular rim close to the anterior inferior iliac spine. More centrally localized cartilage lesions were present in 47% of cadavers and in 28% of patients. The majority of femurs (cadavers) (80%) showed reduced narrowing at the anteromedial femoral head-neck junction causing impingement against the corresponding acetabular rim or squeezing of this area into the joint during flexion, whereas the femoral head cartilage appeared normal in all but one hip. Acetabular rim degeneration is a constant finding in the aged hip, which seems to be triggered by femoroacetabular impingement. PMID- 12897612 TI - Gender differences in surface rolling and gliding kinematics of the knee. AB - The purpose of the current study was to determine whether knee surface rolling and gliding kinematics differed between genders during open and closed kinetic chain movement conditions. Eleven unimpaired adults (six men and five women) participated in this study. Sagittal plane path of instant center of rotation measurements were obtained with videographic motion analysis and applied to a mathematical knee model from which joint surface rolling and gliding kinematics were obtained. In addition, normalized electromyographic data were collected from subjects' quadriceps and hamstring muscles. During closed kinetic chain knee extension, as the knee approached terminal extension, female participants showed significantly greater relative joint surface gliding than male participants. Female participants also extended the knee in the closed kinetic chain with less relative hamstring activity than males. The relationship between joint surface gliding and relative hamstring activity in females during closed kinetic chain knee extension may explain, in part, the greater incidence of noncontact anterior cruciate ligament injury that occurs in females. PMID- 12897613 TI - In vivo three-dimensional patellar tracking on the femur. AB - In vivo three-dimensional patellar tracking under weightbearing conditions was investigated with the principal reference axes of the femur in the coronal and axial planes, using a biplanar image-matching technique. Three-dimensional knee models of eight healthy volunteers were constructed using computed tomography scanning. Projection images of the models were fitted onto anteroposterior and lateral radiographs of the knees at hyperextension and at every 15 degrees from 0 degrees to 120 degrees flexion. Knee motion then was reconstructed on a computer. Patellar tracking during knee flexion was described simply with a medial jerk shift of 8 mm in early flexion until 30 degrees and a linear tracking with minimal mediolateral translation. The linear tracking portion was located laterally 5 mm from the mediolateral center of the femoral condyles. On average, the direction of this linear tracking was almost perpendicular to the distal condylar line in the coronal plane and perpendicular to the posterior condylar axis in the axial plane. These results help improve the understanding of patellofemoral kinematics and provide useful information for the design and positioning of the prostheses used in total knee arthroplasty. PMID- 12897614 TI - Bowing fractures of the forearm in children: a long-term followup. AB - Eleven children with forearm bowing fractures were followed up after 80 months. They were evaluated specifically for pain, function, and cosmesis. Remodeling was assessed by comparing radiographs. The mean age when the trauma occurred was 7 years 5 months. Forces longitudinal to the forearm axis are considered responsible for this injury. In six patients the deforming force was longitudinal, but in two other patients it might have been transversal. In three patients the mechanism of injury was unknown. The initial diagnosis was missed three times. Reduction was cumbersome in five of seven patients. In nine patients a residual curve was accepted. This resulted in a unique opportunity to evaluate the spontaneous remodeling. After 6 years of age, remodeling was less than generally accepted. From this it can be concluded that a transversal force is also capable of causing these injuries, that remodeling is less than generally accepted, and that reduction under general anesthesia should be considered in patients older than 6 years who have a cosmetically unacceptable bowing deformity that is greater than 10 degrees. PMID- 12897615 TI - Second cervical vertebrae pedicle fractures versus synchondrosis in a child. AB - Fractures of the second cervical vertebrae (C2) can be difficult to diagnose in young children. A 17-month-old infant sustained bilateral C2 pedicle fractures after minor trauma. The initial lateral radiograph showed pedicle defects, but even with computed tomography and a bone scan with single photon emission computed tomography this could not be differentiated from a congenital abnormality. One week after injury, repeat radiographs showed spondylolisthesis confirming the defects were fractures. The fractures were treated by closed reduction and a halo vest and healed uneventfully. This child is among a small number of patients with bilateral C2 pedicle fractures that occur predominantly in young children. Diagnosis can be difficult even with a high index of suspicion. PMID- 12897616 TI - Chemotherapy decreases epiphyseal strength and increases bone fracture risk. AB - To establish the effect of three frequently used chemotherapeutic agents in childhood cancer on the skeleton, growing male Wistar rats were studied. Treatment with doxorubicin, methotrexate, and cisplatin reduces the proximal tibial growth plate shear strength because of a decreased surface area and maximum shear stress. After treatment the bone fracture risk of the tibia and femur is increased because of decreased bending resistance. Doxorubicin and cisplatin reduce the maximum shear stress of the proximal tibial growth plate, none of the chemotherapeutic agents inhibit bone mineralization. These effects are caused by treatment-induced malnutrition and the accompanying weight reduction and a direct effect of the chemotherapeutic agents on the skeleton. The current study confirmed the importance of preventing malnutrition during chemotherapeutic treatment in view of possible skeletal complications. During followup of children treated with chemotherapy, attention should be given to signs and symptoms suggestive of such complications. PMID- 12897617 TI - Peripheral primitive neuroectodermal tumor after radiotherapy. AB - A 41-year-old man had a peripheral neuroectodermal tumor develop at the distal third of the fibula 4 years after radiotherapy for relapsed villonodular synovitis. This type of sarcoma usually is classified into the heterogeneic group of small round-cell bone tumors as a subdivision of Ewing's sarcomas. The immuno staining positivity of the neoplastic cells for the neuron-specific enolase allowed the authors to make the diagnosis of a tumor with neuroectodermal origin. When the histologic study confirmed the diagnosis, the patient was treated with chemotherapy, surgical excision of the tumor, and adjuvant radiotherapy. Radiotherapy is thought to be involved in the genesis of osteogenic sarcomas as it has been shown in several reports, but there is no evidence in the literature of a peripheral neuroectodermal tumor developing after radiotherapy. PMID- 12897618 TI - Detection of biomaterial-associated infections in orthopaedic joint implants. AB - Biomaterial-associated infection of orthopaedic joint replacements is the second most common cause of implant failure. Yet, the microbiologic detection rate of infection is relatively low, probably because routine hospital cultures are made only of swabs or small pieces of excised tissue and not of the surfaces of potentially infected implants. Joint replacements from patients in whom infection was suspected, after clinical, radiologic, and biochemical examinations, were used in this study. The aim of the current study was to compare the detection rate of infection in total joint replacements based on cultures of the excised tissue and scrapings from the biomaterial surface. Joint prostheses were retrieved from 22 patients requiring orthopaedic revision surgery because of suspected infection of their prostheses. Routine hospital culturing of tissue only showed bacterial growth in nine patients (41%). However, after prolonged culturing, bacterial growth was observed in 14 patients (64%), whereas extensive culturing of scrapings from the biomaterial surface indicated bacterial growth in 19 of the 22 patients (86%). In addition, confocal laser scanning microscopy enabled observation of biofilm bacteria on the surfaces of the explanted prostheses. Diagnosis in orthopaedic revision surgery should consider using a microbial or microscopic analysis of the surface of an explanted prosthesis, where the biofilm mode of growth firmly anchors and protects the infecting organisms. Improved detection of infection by analysis of the implant surface is expected to yield ameliorated therapy and a reduced need for revision surgery. PMID- 12897619 TI - Effect of extracorporeal shock wave therapy on osteoblastlike cells. AB - Extracorporeal shock wave therapy has been used increasingly in musculoskeletal disorders although its biologic mechanisms are not understood completely. The current study evaluated the effects of extracorporeal shock wave therapy on human osteoblastlike cells by using an electrohydraulic shock wave generator and comparing three energy levels. (Group A, 14 kV and 0.15 mJ/mm2; Group B, 21 kV and 0.31 mJ/mm2; Group C, 28 kV and 0.40 mJ/mm2; Control Group, no energy) and two total impulses (500, 1000) for each level. At the end of treatment, a reduction by approximately 76% was observed in Group C cell number versus basal value when compared with the other groups. Viability, biochemical activity, and gene expression of cultured cells were evaluated 24 and 48 hours after treatment. The viability test showed a decrease in Group C viability of approximately 54% at both culture times as compared with the other groups. Significant increases in nitric oxide, osteocalcin, and transforming growth factor-beta1 production ranging from 10% to 35% were found in Group A. All treated groups had lower C terminal procollagen Type I values than the Control Group, but important increases were observed between 24 and 48 hours in all groups except Group C. This particular finding reveals that osteoblast differentiation in Group A is enhanced strongly during the first 24 hours after exposure leading after another 24 hours to an increase in C-terminal procollagen Type I production and consequently in bone matrix deposition. The current study showed that one of the most important aspects to be considered is not the total number of impulses used, but the energy level of the shock waves, therefore confirming that extracorporeal shock wave therapy has a dose-dependent initial destructive effect on cells when the selected energy is higher than 21 kV. PMID- 12897620 TI - An evaluation of human demineralized bone matrices in a rat femoral defect model. AB - The osteoconductive and osteoinductive potential of two human allogeneic demineralized bone matrix putties were compared in a critical-sized athymic rat femoral defect model. Defects were treated with (1) a demineralized bone matrix in a hyaluronic acid carrier, (2) a demineralized bone matrix in a glycerol carrier, (3) a hyaluronic acid carrier alone, or (4) with no implant. Radiographic examinations and histologic analyses were done at 4, 8, and 16 weeks postoperatively. Eight of the 48 defects treated with a demineralized bone matrix and none of the 36 surgical controls showed complete radiographic healing by 16 weeks and no statistically significant difference between the radiographic scores for the two demineralized bone matrix preparations was found. On histologic review, both preparations of demineralized bone matrix had passive remineralization. The largest foci of endochondral ossification were seen in limbs treated with a demineralized bone matrix in a hyaluronic acid carrier. The 8-mm rat femoral defect allows for stringent assessment of the osteoinductive potential of bone graft substitutes. Hyaluronic acid and glycerol are viable carriers for demineralized bone matrices. As both de-mineralized bone matrices tested provided an adequate osteoconductive matrix and showed some, although limited, osteoinductive capacity, these materials should be used in clinical practice only as bone graft extenders or enhancers. PMID- 12897621 TI - Healing of bone-tendon junction in a bone trough: a goat partial patellectomy model. AB - Bone-tendon junction healing in a bone trough was investigated in a goat partial patellectomy model. Histologic evaluation and biomechanical tests were done at 6, 12, and 24 weeks. Irregular fibrous tissue seen at the healing bone-tendon junction at 6 weeks gradually assumed longitudinal alignment and remodeled toward a direct bone-tendon junction. Type III collagen deposition was diffuse at 6 weeks, but became localized to the healing interface at 12 weeks. Thickness of newly formed bone increased progressively with time. Bridging collagen fibers were formed at the junction, with fibrochondrocytic cells and a basophilic tidemark detected at 24 weeks. The trabecular line remained discontinuous and there was no safranin O uptake. Most specimens failed at the junctions under tensile loads. The ultimate failure stress increased from 4.78 +/- 0.50 N/mm2 at 6 weeks to 7.99 +/- 0.33 N/mm2 at 24 weeks (mean +/- standard error of the mean), only reaching 15% of normal. Cartilage from the articular cut surface extended into the healing interface, later forming an area of fibrocartilage with densely packed collagen fibers aligned along the direction of force, containing proteoglycans. Cartilage may enhance restoration of a transition zone in bone tendon junction healing. The sequence of events outlined formed a basis to guide clinical practice regarding bone-tendon junction reattachment. PMID- 12897622 TI - Teaching orthopaedics on the run: tell me the story backward. AB - At times clinical teaching can become too passive and the cognitive level of discussions with residents may be inappropriately low. Busy outpatients clinics and emergency departments might be considered hostile educational environments that do not lend themselves well to scholarly discussions and intense teaching sessions. Paradoxically, these same areas are where attendings and residents may spend considerable time, and consequently, significant training opportunities may be lost. This situation demands a teaching tactic that is simple, time efficient, and cognitively challenging. Such a teaching tactic, referred to as Tell Me The Story Backward, is presented. PMID- 12897623 TI - Painful patella in a 64-year-old woman. PMID- 12897624 TI - A 29-year-old man with buttock pain. PMID- 12897625 TI - Low back pain in a 13-year-old girl. PMID- 12897626 TI - Public reporting of healthcare data. PMID- 12897627 TI - Cardiac rehabilitation with nurse care management and telephonic interactions at a community hospital: program evaluation of participation and lipid outcomes. AB - Cardiac rehabilitation (CR) can help patients with heart disease control some risk factors, limit new coronary artery lesions, and decrease death rates. However, participation rates in CR are low. Using a descriptive design, we evaluated a modified CR program in which nurse care managers used telephonic communication with patients in their homes by comparing it to a traditional CR program in a hospital setting. Using multivariate analysis we compared the patient cohorts eligible for each of the programs, and program participants to the nonparticipants for each program. Compared to traditional CR, the modified CR program with nurse care management was associated with significantly improved participation rates (11% vs. 22%) and the apparent overcoming of several well described barriers to CR participation (distance from the hospital and domestic isolation). Risk factor management, including testing of serum lipids and achieving goals for lipid reduction, for participants in both CR programs was superior to risk factor management for nonparticipants. PMID- 12897628 TI - Shifting the healthcare paradigm: the case manager's opportunity and responsibility. AB - Recent shifts in the healthcare system reflect a shift in thinking from sickness to wellness. This article reviews the paradigm shift, incentive system, and issues related to cost and quality. The role of case management in facilitating this transition is discussed. The article concludes with a discussion of the challenge to use nursing's unique disciplinary perspective that is focused on health to develop new approaches and strategies and the opportunities for case management to expand its scope of practice. PMID- 12897629 TI - Counteractions to the paradigm shift. AB - While the benefit of a health-focused paradigm is clear theoretically, the transition is tortuous. This article reviews the difficulties of implementing the paradigm shift presented in the preceding article. This article presents counteractions, clarifies underlying assumptions, presents the need for collaborative action, and highlights the potential contribution of case management to the paradigm shift. PMID- 12897630 TI - Predictive modeling and proactive care management: part 1. PMID- 12897631 TI - Nursing home compare: web site offers critical information to consumers, professionals. AB - The U.S. Department of Health and Human Services has launched a Nursing Home Quality Initiative that provides nursing home consumers and healthcare professionals with publicly reported quality information about every Medicare- or Medicaid-certified nursing facility in the United States. The initiative also offers assistance to nursing homes in their continuing efforts to improve the quality of the care they offer. This article provides an overview of that initiative, and discusses information available on the Nursing Home Compare Web site that case managers will find useful for helping clients in their nursing home decision-making process. PMID- 12897632 TI - Declining medication requirement in some patients with advanced Parkinson disease and dementia. PMID- 12897633 TI - Hypothermia and phenytoin toxicity. PMID- 12897634 TI - Transient neurologic deficits associated with carbamazepine-induced hypertension. AB - Carbamazepine is a well-established, effective treatment of complex partial seizures and is well tolerated in most patients. The adverse effects of carbamazepine include aplastic anemia, agranulocytosis, pancytopenia, bone marrow depression, thrombocytopenia, cardiac conduction abnormalities, congestive heart failure, and peripheral edema. Hypertension or hypotension has also rarely been documented in patients with either therapeutic or toxic blood levels of carbamazepine. It is possible that carbamazepine-induced hypertension in those with therapeutic blood levels is rarely seen because most of the patients who begin treatment are young and do not have baseline hypertension. The authors describe a patient of African-American descent with a history of controlled essential hypertension who developed severe uncontrolled hypertension when started on carbamazepine. Treatment with additional antihypertensive medications did not reduce his blood pressure. In addition, he developed two episodes of transient neurologic deficits, the symptoms of which consisted of dysarthria, vertigo, and unstable gait. A substantial reduction of his carbamazepine dose resulted in the control of his blood pressure and no recurrence of his symptoms. PMID- 12897635 TI - Drug rash with eosinophilia and systemic symptoms secondary to phenobarbitone. AB - The authors describe a case of a rare, potentially fatal multisystem reaction, recently called drug rash with eosinophilia and systemic symptoms (or DRESS) resulting from phenobarbitone. They highlight the main characteristics of this syndrome and the diagnostic difficulties. PMID- 12897636 TI - Mirtazapine in L-dopa-induced dyskinesias. AB - Mirtazapine is a novel antidepressant with a pharmacologic profile (alpha-2 antagonist, 5HT-(1A) agonist, and 5HT-(2) antagonist) that renders it potentially useful for l-dopa-induced dyskinesias. Drugs with 5HT-(1A) agonistic activity, such as buspirone and tandospirone, have been reported to be effective in reducing l-dopa-induced dyskinesias. Furthermore, 5HT-(2) antagonism may, by reducing substantia nigra pars reticulata hyperactivity, play a role in the improvement of Parkinsonian symptoms and l-dopa-induced dyskinesias, as has been observed with ritanserin, a 5HT-(2) antagonist. Alpha-2 antagonists, such as idazoxan, have recently also been reported to improve l-dopa-induced dyskinesias. The authors investigated the antidyskinetic properties of mirtazapine by designing an open-label study of 20 Parkinsonian patients with l-dopa-induced dyskinesias. Mirtazapine proved to be moderately effective in reducing l-dopa induced dyskinesias, either alone or in association with amantadine. Mirtazapine may be of use in patients who do not respond or are intolerant to amantadine. PMID- 12897637 TI - The impact of prolactin elevation with antipsychotic medications on subjective quality of life in patients with schizophrenia. AB - In this cross-sectional study, the author tested the hypothesis that prolactin elevation with antipsychotic medications was associated with low subjective quality-of-life scores in patients with schizophrenia. The subjects were 42 male inpatients meeting Diagnostic and Statistical Manual of Mental Disorders, 4th edition, criteria for schizophrenia on typical antipsychotics. No correlations were found between prolactin or testosterone and the 3 subscales in the Japanese version of the Schizophrenia Quality of Life Scale. Multiple regression analyses showed total variance in the ratings of 3 subscales in the Japanese version of the Schizophrenia Quality of Life Scale as follows: Brief Psychiatric Rating Scale (BPRS) anxiety/depression factor, dosage of antipsychotics and BPRS hostile/suspiciousness factor in the psychosocial subscale (adjusted R2 = 0.394), BPRS anxiety/depression factor and dose of antipsychotics in the motivation/energy subscale (adjusted R2 = 0.475), and dose of antipsychotics and BPRS anergia factor in the symptoms/side effects subscale (adjusted R2 = 0.206). The results did not support the hypothesis. PMID- 12897638 TI - Pharmacokinetic studies of single and multiple oral doses of fampridine-SR (sustained-release 4-aminopyridine) in patients with chronic spinal cord injury. AB - Fampridine (4-aminopyridine) is a potassium channel blocking agent that restores conduction in demyelinated axons and improves neurologic function in patients with chronic spinal cord injury (SCI). Based on the pharmacokinetic profile of orally administered fampridine, multiple daily doses (4 or more) would need to be taken to sustain its therapeutic effects. Two studies were conducted to determine the pharmacokinetics and safety profile of an oral, sustained-release (SR) formulation of fampridine (fampridine-SR, 10-25 mg) administered as a single dose (n = 14) and twice daily for 1 week (n = 16) in patients with chronic, incomplete SCI. Mean plasma concentrations and area under the plasma concentration-time curve were proportional to the dose administered, whereas other pharmacokinetic parameters were independent of dose. Fampridine-SR was absorbed slowly (peak plasma concentration shortly after dosing, 2.6-3.7 hours) and eliminated (plasma half-life, 5.6-7.6 hours), and reached steady state after 4 days of twice-daily administration. Fampridine-SR was well tolerated, with only mild to moderate adverse events reported, and no serious adverse events. The extended plasma half life of fampridine-SR allows convenient twice-daily dosing. Clinical trials designed to assess neurologic and functional improvement using fampridine-SR in patients with chronic SCI are currently underway. PMID- 12897639 TI - Selegiline in the treatment of sexual dysfunction in schizophrenic patients maintained on neuroleptics: a pilot study. AB - A double-blind, placebo-controlled crossover study was undertaken in 10 neuroleptic-treated male schizophrenic outpatients to assess the effect of coadministration of selegiline 15 mg/day for 3 weeks on their sexual dysfunction. Selegiline was not found to be effective in improving any domain of sexual functioning despite a significant decrease in prolactin levels (P < 0.05). This study emphasizes the complex nature of sexual dysfunction in schizophrenic treated patients and the need for placebo-controlled trials for this condition. PMID- 12897640 TI - Waiting for ON: a major problem in patients with Parkinson disease and ON/OFF motor fluctuations. AB - The authors compared the two portions of the OFF period in patients with Parkinson disease and response fluctuations: time to ON (the latency from levodopa intake to turning ON) and wearing off (time from termination of the beneficial dose effect until the time when the next dose was taken). Time to ON was more than twice the duration of wearing off. Although underrecognized, time to ON is the major component of total daily OFF. PMID- 12897641 TI - Adjuvant topiramate administration: a pharmacologic strategy for addressing NMDA receptor hypofunction in schizophrenia. AB - N-methyl-d-aspartate receptor hypofunction (NRH) and its downstream consequences, especially excitotoxicity, may explain the progressive psychosocial deterioration and ventriculomegaly observed in at least some patients with schizophrenia. Topiramate has several properties that address downstream consequences of NRH. In this open-label investigation, the authors examined the salutary therapeutic effects of adjuvant topiramate in 12 patients with schizophrenia and schizoaffective disorder. Patients were selected on the basis of the presence of negative symptoms. An optimal dose of topiramate was determined for each patient during a slow 4-week titration process. Patients were maintained on topiramate and their stable antipsychotic medications for 8 weeks, after which topiramate was tapered and discontinued. Patients were followed for an additional 4 weeks on their stable antipsychotic medications. Clinical measures of efficacy (eg, Positive and Negative Syndrome Scale), cognitive measures (eg, verbal fluency, memory), and safety measures (eg, postural sway) were assessed throughout this study. Topiramate administration (average dose, 110.42 mg/day) decreased total scores on the Positive and Negative Syndrome Scale. Topiramate was also associated with a selective and reversible worsening of verbal fluency performance. These results encourage further testing of topiramate and kainate/alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid receptor antagonists in schizophrenia patients and support the heuristic model of NRH. PMID- 12897642 TI - Albuterol improves response to levodopa and increases skeletal muscle mass in patients with fluctuating Parkinson disease. AB - Animal studies indicate that beta(2)-adrenergic receptor agonists enhance transport of levodopa across the blood-brain barrier. Preliminary studies showed improved response to levodopa in patients with Parkinson disease (PD) who were given albuterol as adjunctive therapy. Beta(2)-adrenergic agonists may offer additional benefits to PD patients via their skeletal muscle anabolic effects, particularly those who experience decreased muscle strength and weight loss. Nondemented, fluctuating PD patients receiving levodopa but not experiencing severe dyskinesias underwent the following tests at baseline and 14 weeks after treatment with albuterol sulfate (4 mg four times a day, orally): Unified Parkinson's Disease Rating Scale motor, tapping, and stand-walk-sit tests every 30 minutes between 8 am and 5 pm; body composition analyses using whole-body plethysmography and computed tomography of the thigh; muscle strength tests; and the Parkinson's Disease Questionnaire (PDQ-39). Results were analyzed using paired t-tests (2 tailed), repeated-measures analysis of variance, and the Wilcoxon signed-rank test. Seven of 8 enrolled patients completed the study; 1 patient withdrew because of headache and anxiety. The area under the curve for all-day UPDRS motor scores improved by 9.8 +/- 9.1% (mean +/- standard deviation; P < 0.05) and tapping improved by 7.6 +/- 8.1% (P < 0.05). The effect was more pronounced when only the response to the first levodopa dose (area under the curve, 8-11 am) was analyzed: 13.0 +/- 9.8% and 9.8 +/- 9.6% respectively. Thigh muscle cross-sectional area increased significantly as measured by computed tomography (5.3 +/- 3.2%, P < 0.01), as did fat-free mass by whole-body plethysmography combined with total-body water determination (9.5 +/- 2.9%, P < 0.05). There was no significant improvement in the stand-walk-sit test, muscle strength tests, other UPDRS sections, daily OFF time, or PDQ-39. Four patients were rated as having a mild global improvement (+1 point) on a -3 to +3-point scale, and 3 of them chose to continue albuterol beyond the termination of the study. The mean heart rate increased from 78.3 +/- 9.3 beats/minute to 85.6 +/- 8.7 beats/minute (P < 0.05). No laboratory abnormalities or electrocardiographic changes were induced by albuterol in any subject. This open-label pilot study suggests that albuterol increases muscle mass and improves the therapeutic response to levodopa in patients with fluctuating PD. A double-blind, placebo controlled study is needed to confirm the effects and safety profile of beta(2) agonists in PD. PMID- 12897643 TI - Pressor response to intravenous tyramine in healthy subjects after safinamide, a novel neuroprotectant with selective, reversible monoamine oxidase B inhibition. AB - Safinamide is a novel neuroprotectant combining sodium and calcium channel blocking properties with selective, reversible monoamine oxidase type B (MAO B) inhibition. Phase 1 studies have demonstrated that in healthy volunteers, the ED50 (a dose that inhibits enzyme activity by 50% in 50% of treated subjects) for MAO B inhibition is 87.5 microg/kg/day orally, and that no MAO A inhibition occurs after 10-mg/kg oral dosing. To assess the risk of inducing the "cheese effect," the effect of safinamide and placebo on the pressor response to tyramine was investigated in a group of healthy male volunteers. The study was an open, single-dose placebo-controlled trial with the 2 treatments in sequence. An increase of 30 mm Hg systolic blood pressure was obtained by intravenous tyramine administered by 0.5-mg incremental boluses injected at 15-minute intervals. The amount of tyramine necessary to achieve such a blood pressure increase was the same after the safinamide 2-mg/kg oral load compared with placebo. These results suggest that dietary restrictions for food with high tyramine content should not be required under safinamide treatment. PMID- 12897644 TI - Metabolite changes resulting from treatment in children with ADHD: a 1H-MRS study. AB - Previously the authors noted an increase in glutamatergic tone in children with attention deficit hyperactivity disorder compared with age- and gender-matched control subjects. In this study they examine the effect of treatment on metabolite concentrations. Fourteen children with attention deficit hyperactivity disorder were investigated medication free and after treatment, using proton magnetic resonance spectroscopy. In the prefrontal cortex and striatum, metabolite peaks of N-acetyl-aspartate, glutamate/glutamine/gamma-aminobutyric acid, creatine/phosphocreatine, and choline compounds were measured, and ratios of the peaks were calculated and compared before and after treatment. The glutamate/glutamine/gamma-aminobutyric acid-to-creatine/phosphocreatine ratio decreased significantly in the striatum. No other metabolites demonstrated any change in response to medication. These findings suggest that glutamate may be involved in treatment response in attention deficit hyperactivity disorder, especially in the striatum. PMID- 12897645 TI - Reverse of the differential uptake intensity of Tc-99m MIBI and Tc-99m V-DMSA by multiple myeloma lesions in response to therapy. AB - PURPOSE: The objective of this study was to compare the uptake changes of Tc-99m 2-methoxy isobutyl isonitrile (MIBI) and Tc-99m pentavalent dimercaptosuccinic acid (V-DMSA) in multiple myeloma (MM) lesions in response to high-dose chemotherapy (HDC). MATERIALS AND METHODS: The authors compared Tc-99m MIBI and Tc-99m V-DMSA scans before and after HDC in a patient with focal MM lesions without amyloidosis who had received previous standard chemotherapy as well. RESULTS: HDC had the effect of eliminating all Tc-99m MIBI uptake in the lesions. Tc-99m V-DMSA uptake was increased in lesions presenting significant initial Tc 99m MIBI uptake. In 1 particular lesion that demonstrated this phenomenon, magnetic resonance showed necrosis of the area of MM. CONCLUSION: The authors consider that the effect of increasing Tc-99m V-DMSA uptake in the absence of an increase in viable plasma cells possibly reflects the treatment-generated inflammatory and fibrotic changes and not necessarily viable tumor tissue. Exclusive focal Tc-99m V-DMSA uptake in this clinical setting could be considered as a sign of effectively treated lesions and not a sign of deterioration. PMID- 12897646 TI - Diagnosis of labia metastasis by F-18 FDG PET and CT fusion imaging in sarcoma follow-up. AB - ABSTRACT A 32-year-old woman with a history of pelvic sarcoma underwent whole body fluorine-18 fluorodeoxyglucose positron emission tomography (PET) and computed tomographic (CT) fusion imaging for restaging after primary surgery and adjuvant combined radiation and chemotherapy. Interpreting the PET images alone showed an area of focal tracer accumulation in the pelvic floor and was interpreted as being secondary to urine contamination. Image fusion with fully coregistered CT scans showed a localized site of increased uptake from a metastasis to the labia, which was confirmed by biopsy. The presented case illustrates the usefulness of combined PET/CT fusion imaging to overcome some pitfalls of PET imaging alone (eg, differentiating malignant tumor from pooling of tracer in the genitourinary tract or contamination). PMID- 12897647 TI - Hepatobiliary scintigraphy in liver transplant patients: the "blind end sign" and its differentiation from bile leak. AB - PURPOSE: Hepatobiliary scintigraphy is commonly performed on liver transplant patients to exclude biliary complications such as obstruction or leak. Biliary reconstruction in these patients is performed using either a direct duct-to-duct anastomosis (choledochocholedochostomy) or a biliary-enteric anastomosis (Roux-en Y hepaticojejunostomy). The authors have observed a finding in scans performed on the latter group of patients that often simulates a bile leak. They postulate this finding to represent retention of activity in the blind end of the Roux limb (the "blind end sign"). The purpose of this study was to determine characteristics of this blind end activity, which may help differentiate it from true bile leaks. METHODS: Two nuclear medicine physicians reviewed 36 Tc-99m mebrofenin hepatobiliary scans (31 patients). Biliary reconstruction was performed in 25 patients with a Roux-en-Y hepaticojejunostomy, and in 6 patients with a choledochocholedochostomy. Each scan was evaluated regarding whether there was focal, persistent retention of activity in the anastomotic region, and, if so, various features of the activity were noted. Activity was concluded to represent a true bile leak based on subsequent surgical confirmation, and if there was no such confirmation, the activity was concluded to represent blind end retention in patients with Roux-en-Y hepaticojejunostomies. RESULTS: There were 8 total examples of blind end activity and 3 examples of bile leak activity. Useful discriminators between the 2 types of activity included the following: fluctuation in size and/or intensity with time (8 of 8 blind end cases vs. 0 of 3 bile leak cases); size comparable with bowel (8 of 8 vs. 1 of 3); tubular (4 of 8 vs. 0 of 3), round (4 of 8 vs. 1 of 3), or irregular (0 of 8 vs. 2 of 3) shape; and movement into bowel (3 of 8 vs. 0 of 3). Delayed imaging was performed in 3 cases at 1.5 hours, but was only conclusive in 1 case in which diffuse extravasation was seen consistent with bile leak. CONCLUSION: Retention of activity in the blind end of the Roux limb in patients with Roux-en-Y hepaticojejunostomies may simulate bile leak, but certain typical features, as well as delayed imaging, may aid in distinguishing the two. PMID- 12897648 TI - Imaging osteomyelitis with Tc-99m-labeled antigranulocyte antibody Fab' fragments. AB - PURPOSE The purpose of this study was to evaluate the diagnostic value of Tc-99m labeled antigranulocyte antibody fragments in the diagnosis of osteomyelitis. MATERIALS AND METHODS Thirty immunoscintigrams were evaluated retrospectively with 740 MBq (20 mCi) Tc-99m-labeled antigranulocyte antibodies. The final diagnoses were confirmed by histology, magnetic resonance imaging, computed tomography, and clinical follow-up. RESULTS In the retrospective analysis, 20 of 30 patients (67%) demonstrated a true-positive result. Three of 30 patients (10%) had a false-positive result. A false-negative result was found in a diabetic patient with a perforating ulcer of the foot. Six of 30 patients had a true negative result. A high sensitivity of 95%, a relatively high specificity of 67%, and a high diagnostic accuracy of 86% were present in this study. CONCLUSION Tc 99m labeled monoclonal antibody-Fab' fragments are suitable for the detection of osteomyelitis. Its clinical application is simple. Its use guarantees a reliable and accurate diagnostic result just 1 to 2 hours after injection, making a late scan unnecessary. Coxarthrosis or hyperostosis can lead to false-positive results. A perforating ulcer of the foot may result in a false-negative conclusion. PMID- 12897649 TI - Thymic uptake of radiopharmaceuticals. AB - The thymus, as the site of T-cell differentiation, plays a critical role in the development of the immune system. The gland increases in weight until puberty and then is slowly replaced by fat. Various radiopharmaceuticals can localize to the thymus before it involutes. This pictorial presentation reviews the appearance and proposed mechanisms of thymic uptake of Ga-67 citrate, F-18 fluorodeoxyglucose, radioiodine, and In-111 pentetreotide. PMID- 12897650 TI - Erythema nodosum associated with diffuse, large B-cell non-Hodgkin lymphoma detected by FDG PET. AB - A patient is described with non-Hodgkin lymphoma and erythematous skin nodules suspected to be erythema nodosum. The patient underwent serial fluorodeoxyglucose (FDG) positron emission tomography (PET), which demonstrated normalization of FDG uptake by the lymphoma after 2 cycles of chemotherapy, but there was new abnormal uptake involving the subcutaneous tissues of the lower extremities. A typical skin lesion was sampled and showed the appearance of erythema nodosum with no evidence of lymphoma. The FDG uptake gradually diminished on serial PET imaging after treatment with nonsteroidal antiinflammatory drugs. In view of the recognized association of erythema nodosum with malignancy and the differential rate of response to chemotherapy, the lesions of erythema nodosum may be a source of a false-positive PET interpretation, and histologic assessment should be considered. PMID- 12897651 TI - Unrecognized renal transplants as a potential source of false-positive interpretation of FDG PET. AB - Renal transplantation has become an effective therapy for patients with late stage renal disease. Fluorodeoxyglucose (FDG) positron emission tomography (PET) is accepted as an important diagnostic technique in the evaluation of suspected or known malignancies or other disorders in the day-to-day practice of medicine. Because FDG is excreted from the kidneys into the urine, unrecognized renal transplants can appear as malignant lesions. Familiarity with the clinical history is a prerequisite in the correct interpretation of FDG PET images in this setting. In addition, FDG PET images should be correlated with anatomic images when such studies are available. When neither clinical history nor anatomic images are available, a combination of "abnormal" activity in the pelvis and absence of normal renal activity should raise suspicion of the existence of a renal transplant. PMID- 12897652 TI - Clinical significance of rapid (accelerated) gastric emptying. AB - PURPOSE: The purpose of this study was to determine the significance and frequency of rapid gastric emptying (RGE) in clinical practice and to examine its relationship to age, gender, and presenting symptoms. METHODS: The review included 750 patients who underwent scintigraphic gastric emptying studies (GES) within the past 11 years. In 80 patients with RGE, the mean gastric emptying time (GET) in female and male patients was compared with study gender differences. Similarly, the mean GET in young and old patients was compared with study age differences. The presenting symptoms and conditions indicating GES were noted. RESULTS: RGE was found in 80 of 750 study patients (10.7%). RGE was equally common in both sexes. Younger patients (mean age, 34.3 +/- 9.5 years) and older patients (mean age, 67.3 +/- 11.6 years) had similar mean GETs. Most patients with RGE presented with symptoms that mimicked gastroparesis. CONCLUSIONS: RGE has no relationship to age or gender. RGE may be representative of early dumping syndrome in most patients. GES help make a clear distinction between early dumping syndrome and gastroparesis, which can present with similar symptoms. PMID- 12897653 TI - Fractured osteophyte demonstrated on SPECT and computed tomography. AB - ABSTRACT We present an interesting case of a fractured osteophyte causing back pain that was demonstrated both on bone single photon emission computed tomography (SPECT) and computed tomography (CT). The magnetic resonance images, thoracic anterior spine plain radiograph, whole-body bone scan passes, and thoracic spot view were not impressive. Bone SPECT was the impetus for getting the CT scan. The CT scan not only demonstrated the osteophyte but a pseudarthrosis that was probably causing the pain. If it were not for the positive SPECT bone scan, the CT scan would not have been ordered after unimpressive magnetic resonance imaging. PMID- 12897654 TI - Fluorodeoxyglucose positron emission tomography excludes pericardial metastasis by recurrent lung cancer. AB - Fluorodeoxyglucose (FDG) positron emission tomography (PET) has been shown to be useful in the diagnosis and staging of a variety of malignancies. Because of its high sensitivity, FDG PET frequently detects malignant lesions that are not demonstrated clearly by anatomic imaging modalities. FDG PET usually has high negative predictive value and, therefore, negative studies are highly suggestive of a benign process. The authors present a patient in whom transesophageal echocardiography and magnetic resonance imaging described pericardial metastasis from a recurrent lung cancer, which on FDG PET was shown correctly to suggest benign scar tissue. PMID- 12897655 TI - Incidental detection of pigmented villonodular synovitis on FDG PET. AB - 18 Fluorine fluorodeoxyglucose (FDG) has been used extensively in positron emission tomographic (PET) imaging for oncology. FDG is a glucose analog demonstrating enhanced uptake in the majority of malignant as well as granulomatous lesions. Although a few cases of pigmented villonodular synovitis using bone scanning agents Tl-201 chloride, Ga-67 citrate, and Tc-99m pentavalent dimercaptosuccinic acid have been published, to our knowledge FDG PET imaging of pigmented villonodular synovitis has not. PMID- 12897656 TI - F-18 FDG PET demonstration of sclerosing mesenteritis. AB - Sclerosing mesenteritis is a rare, combined inflammatory and fibrotic process of unknown etiology. Detailed cross-sectional imaging of this entity has been reported with computed tomography. The author presents an additional case of sclerosing mesenteritis demonstrated on fluorine-18 fluorodeoxyglucose positron emission tomography with multidetector computed tomographic correlation. PMID- 12897657 TI - The usefulness of a repeat study for differentiating between bowel activity and local tumor recurrence on FDG PET scans. AB - The importance of fluorodeoxyglucose (FDG) positron emission tomography (PET) in the evaluation of colorectal cancer is well established. Activity in the rectosigmoid region on an FDG PET scan may vary significantly, which may occasionally lead to difficulties in interpretation when assessing local recurrence of colorectal cancer. The authors present a case in which a repeat PET study performed after an enema 1 week later was useful for differentiating between normal bowel activity and local tumor recurrence. This repeat study obviated the need for surgical intervention, although the role of the enema in this case is unclear. PMID- 12897658 TI - Diffuse bone marrow involvement of Hodgkin lymphoma mimics hematopoietic cytokine mediated FDG uptake on FDG PET imaging. AB - Whole-body fluorine-18 fluorodeoxyglucose (F-18 FDG) positron emission tomography (PET) has been useful in the management of a variety of malignancies. In patients with chemotherapy followed by bone marrow stimulants such as granulocyte colony stimulating factor and granulocyte-macrophage colony-stimulating factor, the bone marrow will have diffuse, increased FDG accumulation. Therefore, diffuse bone marrow FDG uptake is commonly attributable to the effect of hematopoietic cytokines. However, diffuse bone marrow FDG uptake can also be caused by bone marrow involvement by malignancy. The authors report a patient with diffuse bone marrow involvement of Hodgkin disease that appears indistinguishable from hematopoietic cytokine-mediated FDG bone marrow uptake. PMID- 12897659 TI - F-18 FDG PET imaging in gastrointestinal stromal tumor. AB - Gastrointestinal stromal tumor (GIST) is the most frequent mesenchymal malignancy of the gastrointestinal tract. Liver and peritoneum are the most frequent metastatic sites. Surgery is the mainstay of treatment in patients with localized disease. Imatinib mesylate (Gleevec; Novartis Pharmaceuticals, East Hanover, NJ), an inhibitor of tyrosine kinase activity of KIT receptor, has been shown to be an effective treatment in metastatic or unresectable disease. Follow-up of patients treated with Gleevec is controversial. The authors present a case of GIST that showed near-total response with Gleevec on fluorodeoxyglucose positron emission tomography. They suggest positron emission tomography as a useful imaging modality for the monitoring of therapy response in GIST tumors treated with Gleevec. PMID- 12897660 TI - Ureteroinguinal hernia. AB - A ureteroinguinal hernia was demonstrated on both a Tc-99m mercaptoacetyltriglycine (MAG3) renal scan and a computed tomographic scan in an 81-year-old man with suspected urinary tract infection and a prior history of transurethral resection of the prostate. Ureteral hernias are relatively rare, although the ureteroinguinal hernia, found in elderly men on the right side, is the most common type. Ureteral hernias may be asymptomatic but can be present in patients with symptoms of obstructive uropathy. PMID- 12897661 TI - Abnormal MAG3 renal scintigraphy resulting from dehydration. AB - Routine Tc-99m mercaptoacetyltriglycine (MAG3) renal scintigraphy in a 64-year old quadriplegic woman revealed normal blood flow with abnormal, continual parenchymal uptake and minimal excretion into the pelvocaliceal collecting systems and urinary bladder. Because there was no clinical reason to suspect acute tubular necrosis, renal artery stenosis, or ureteropelvic junction (UPJ) obstruction, relative dehydration was considered likely. A repeat MAG3 renal scan 6 days later using intravenous fluids to ensure adequate hydration was completely normal. This case highlights the importance of patient preparation to include adequate hydration. PMID- 12897662 TI - Transient aphasia as late-onset seizure after embolization for dural arteriovenous malformation. AB - Tc-99m HMPAO single photon emission computed tomography was performed in a patient who had experienced aphasia for 3 days. The patient had a medical history of subarachnoid hemorrhage and intracranial hemorrhage. Both of these hemorrhages were related to a dural arteriovenous malformation that was embolized after the onset of the cerebral vascular events. Focal hyperperfusion was seen in the left temporal lobe adjacent to a perfusion defect corresponding to the old cerebral hemorrhage. The finding was interpreted as the focus of the patient's epilepsy. His aphasia resolved after the administration of anticonvulsant medication for several days. PMID- 12897663 TI - Obstructed ectopic kidney mimicking Chilaiditi syndrome on F-18 FDG imaging. AB - Fluorine-18 fluorodeoxyglucose positron emission tomography was obtained in a patient with a duodenal mass. Conventional imaging revealed multiple dilated intestinal loops. Initial impression of positron emission tomographic images was interposition of a loop of bowel between the liver and diaphragm (Chilaiditi syndrome) in an unusual location, with absent right kidney activity. Computed tomography and magnetic resonance imaging of the abdomen showed an obstructed ectopic kidney in this position with a markedly dilated collecting system. There is absence of fluorodeoxyglucose in the markedly dilated renal collecting system resulting from acute obstruction, giving the false appearance of gas-filled bowel loops with an unusual location of the interposition. PMID- 12897664 TI - Recurrent gastrointestinal bleeding diagnosed by delayed scintigraphy with Tc-99m labeled red blood cells. AB - A 56-year-old woman presented with bright-red blood from the rectum. Esophagogastroduodenoscopy revealed mild gastritis. Colonoscopy demonstrated diverticulosis without active bleeding, and in vitro tagged red blood cell scintigraphy was unremarkable. There was no further evidence of bleeding and the patient was discharged home. The patient returned with recurrent bright-red blood from the rectum. Although delayed scintigraphic images seldom demonstrate the site of bleeding, delayed images at 12 hours demonstrated active bleeding near the hepatic flexure in this patient. This was confirmed with selective mesenteric angiography, and was treated with coil embolization of the tertiary branches of the right middle colic artery. PMID- 12897665 TI - Positive leukocyte and negative bone scintigraphy in extensive arthroplasty infection. AB - An 85-year-old male resident of a nursing home presented after a fall with a painful left hip, confusion, and fever. He had multiple medical problems including severe vascular disease and poor vision as a result of macular degeneration. An Austin Moore left hip prosthesis had been inserted for a fractured femoral neck several years before. A septic workup showed a raised leukocyte count but no other focal abnormality. Plain radiography showed a periprosthetic fracture of the left hip. Blood cultures grew beta-hemolytic group A Streptococcus sensitive to penicillin. Bone scintigraphy was thought to be consistent with loosening of the prosthesis without evidence of a recent fracture. Tc-99m leukocyte scintigraphy was markedly abnormal, with extensive soft-tissue uptake suggestive of a large periprosthetic infective collection. This was confirmed at surgery with drainage of 200 mL pus from the left hip, and deep and superficial soft tissues of the thigh. The trochanteric fracture was well granulated and thought to be of long standing. The prosthesis was removed and the patient was treated with appropriate antibiotics with good effect. PMID- 12897666 TI - Is osteoid osteoma an iodophilic lesion?: pathologically proved osteoid osteoma of nasal bone first seen on whole-body iodine-131 scan. AB - The authors present a case of pathologically proved osteoid osteoma that was visualized randomly on an I-131 whole-body scan. Search of the medical literature did not reveal any mention of radioactive iodine uptake by osteoid osteomas. Therefore, the authors concluded that this pathology must be included in the differential diagnostic list of positive findings on I-131 scans. PMID- 12897667 TI - Blunt trauma soft-tissue uptake on skeletal scintigraphy. AB - Muscle and soft-tissue uptake have been described numerous times in the literature in patients undergoing Tc-99m methylene diphosphonate skeletal scintigraphy following trauma. Many of these traumas range from electrical burns, intramuscular injections, sports-related trauma, and overexercising, to actual physical damage resulting in superficial bruising and skin discoloration. In this case the patient presented with continuing low back pain after falling down some stairs. Although the scan was essentially normal for bony trauma, subtle soft tissue uptake was detected that was enhanced with the addition of single photon emission computed tomographic imaging and surface rendering. PMID- 12897668 TI - Gallium-67 imaging in a patient with malignant thyroid lymphoma that spontaneously regressed after subtotal gastrectomy. AB - A patient presented with malignant thyroid lymphoma in association with Hashimoto disease. Abnormal, intense Ga-67 accumulation in the thyroid was seen. The patient underwent subtotal gastrectomy for cancer and the malignant thyroid lymphoma regressed spontaneously. Ga-67 accumulation in the thyroid also decreased. The resected stomach was positive for Helicobacter pylori. The patient achieved complete clinical response after H. pylori eradication. Regression of the malignant thyroid lymphoma might be related to H. pylori eradication and the subtotal gastrectomy. PMID- 12897669 TI - Comparison of I-123 IMP and Ga-67 citrate scintigraphy of malignant melanoma. AB - I-123 iodoamphetamine (IMP) and Ga-67 scintigraphy of malignant melanoma were performed to compare the accumulation of these tracers regarding tumor diameter to determine the site of I-123 IMP accumulation and to evaluate the use of I-123 IMP and Ga-67 in postoperative follow-up. The studies involved 44 patients with a definite diagnosis of malignant melanoma during a 10-year period from April 1991 through March 2001. In this study a very high detection rate of 90.9% was obtained for primary foci with I-123 IMP. With Ga-67, the primary site detection rate was 25%. The results suggest that I-123 IMP is superior to Ga-67 with respect to the detection rate of primary site. PMID- 12897670 TI - Hiatal hernia in iodine-131 scintigraphy: a potential cause of false-positive midline thoracic uptake. AB - There are occasional reports of a hiatal hernia demonstrating thoracic uptake on I-131 scintigraphy. This is a cause of a potential false-positive appearance mimicking a pulmonary or mediastinal metastasis. Hiatal hernia is 1 of a number of causes of false-positive I-131 whole-body scans. Although previous case reports have demonstrated uptake to lie to the left of the midline, this case is slightly unusual in view of the midline appearance of the uptake. Although this appearance could be seen with mediastinal, pulmonary, or prevertebral metastases, hiatal hernia should be considered in the differential diagnosis of uptake in the thoracic midline on I-131 scintigraphy. PMID- 12897671 TI - Abdominal aortic aneurysm demonstrated on renal scintigraphy. AB - A 74-year-old hypertensive woman presented with abdominal discomfort and a pulsatile abdominal mass. Anterior abdominal angiography during cardiac blood pool, and renal scintigraphic imaging demonstrated a large abdominal aortic aneurysm. 1, 2 Before endovascular repair with an aortoiliac endograft, the abdominal aneurysm measured 7.5 x 7.0 cm on abdominal computed tomography. This study demonstrates that a suspected abdominal aortic aneurysm can be confirmed using the addition of anterior abdominal imaging with normal posterior imaging at the time of renal scintigraphy. PMID- 12897673 TI - Diarrhea and wound healing. PMID- 12897674 TI - Partial-thickness burns: identification and management. AB - PURPOSE: To provide an overview of the appropriate evaluation and management of partial-thickness burns. TARGET AUDIENCE: This continuing-education activity is intended for physicians and nurses with an interest in learning about burn wound care. LEARNING OBJECTIVES: After reading the article and taking the test, the participant will be able to: 1. Describe the classification of burn wounds. 2. Identify characteristics of burn wounds and the clinical techniques for diagnosing burn wound depth. 3. Identify the treatment options for partial thickness burns. PMID- 12897675 TI - Pressure ulcer research funding in America: creation and analysis of an on-line database. AB - OBJECTIVE: To systematically collect information on active research grants to characterize pressure ulcer research funding in the United States and to identify potential targets for future research and funding initiatives. DESIGN A descriptive study. MAIN RESULTS: The investigators identified 32 grants, representing 16,444,117 US dollars in research funding. The majority of this funding came from federal sources, including the National Institutes of Health (90%), the Department of Veterans Affairs (7%), the National Institute on Disability and Rehabilitation Research (2%), and the Agency for Healthcare Research and Quality (1%). One quarter of pressure ulcer research grants related to quality improvement. Additional topic areas included risk factors or risk assessment tools (19%), adjunctive therapy (16%), mobilization (13%), and pressure management in foot care for patients with diabetes mellitus (9%). Further grants were in the areas of incidence, assessing tissue damage or healing, support surfaces, dressings and topical agents, nutrition, economic evaluation, and pain. CONCLUSION: The investment in pressure ulcer research is minute compared with pressure ulcer treatment expenditures. Policy makers are urged to encourage increased federal and foundation funding for research concerning pressure ulcers. Researchers are also encouraged to develop well designed proposals to obtain available research funding. Additional research is needed in the areas of pressure ulcer incidence and prevalence, support surface design and use, pain, operative treatment, economic impact, and education strategies for caregivers and patients. PMID- 12897676 TI - Effects of ankle-to-knee external pressures on skin blood perfusion under and distal to compression. AB - OBJECTIVE: To compare the effects of select leg compression pressures on blood perfusion in the skin overlying bone and the skin distal to the leg compression. DESIGN: Blood perfusion of skin overlying the tibia and the foot dorsum was simultaneously measured in 12 healthy subjects using laser Doppler. Each subject's calf was compressed from ankle to knee with an air cast that applied external pressure, ranging from 0 to 40 mm Hg in 10 mm Hg increments. All measurements were initiated after a 20-minute rest interval and were recorded with each subject in a supine position. SETTING: University research center. RESULTS: The main findings show that compression causes a significant decrease in skin blood perfusion (SBF) at both sites, with greater reductions in SBF occurring with increased compression pressure. Greater SBF reductions occurred at the foot than at the directly compressed tibia site. Thus, at the highest pressure (40 mm Hg), the foot SBF reduction (61.6% +/- 13.7%) was greater than at the directly compressed tibial site (33.3% +/- 0.3%), both of which were significantly less than baseline (P <.01). CONCLUSIONS: The present findings and considerable related evidence indicate that lower extremity external compression, within the range of commonly used therapeutic levels, reduces SBF by an amount that depends on the magnitude of the compression pressures. The quantitative difference between the effect of compression on SBF within the compression area, and distal to it, is likely a result of the way that the compression affects arteriolar and venous resistance to blood flow at the 2 sites. From a clinical perspective, it would seem prudent to anticipate that sub-bandage and distal SBF would be negatively affected by compression, with greater effects likely in tissue distal to compression. PMID- 12897678 TI - Cellulitis. PMID- 12897677 TI - 2003 Surgical Dressing Medicare Policy update. PMID- 12897679 TI - Prevalence of human papillomavirus infection among women in Concordia, Argentina: a population-based study. AB - BACKGROUND: Preparing for HPV vaccine programs, studies are needed of HPV infection in different populations. GOAL: The goal was to evaluate HPV prevalence and determinants in Concordia, Argentina. STUDY DESIGN: A stratified random sample of 1786 households was obtained. Consenting women aged > or =15 years were interviewed and underwent examination, including colposcopy. Cells were collected for a Papanicolaou smear and HPV DNA testing with GP5+/6+ primer-mediated PCR EIA. RESULTS: PCR was performed on specimens from 987 women. Prevalence among women reporting no previous sexual activity was 3%, and among sexually active women it was 17.7%, peaking at <25 years of age and decreasing to a minimum at > or =65 years of age. However, low-risk types had similar prevalence (approximately 5%) in all age groups. HPV16 (4.0%), HPV35 (2.6%), and other high risk types were the most common. Almost half of infections were multiple. Younger women initiated sexual activity earlier and had more partners. The main determinants of HPV detection were lifetime number of sex partners and vaginal discharge. CONCLUSION: A clear pattern of decreasing prevalence of HPV with age was observed. This could be explained by development of immunity against specific types over time or related to a cohort effect associated with a recent spread of HPV in this population after recent changes in sexual behavior. PMID- 12897680 TI - Needs assessment for STD/HIV prevention among patients with sexually transmitted diseases in southern China. AB - BACKGROUND AND GOAL: Patients with sexually transmitted diseases (STDs) serve as a bridge population to transmit HIV from higher-risk to lower-risk populations. Our goal was to understand the level of risk behaviors and HIV/STD-related knowledge as well as attitudes among STD patients in China. STUDY DESIGN: An anonymous questionnaire was self-administered by 619 STD patients in Guangzhou, China. RESULTS: About 70% of the males and 18% of the females had had sex with nonregular sex partners in the previous 6 months; the prevalence of consistent condom use was very low (20% for males and 10% for females). Only 20% to 30% of the respondents stated that they would not practice unprotected sex with nonregular sex partners before their STD was cured. Misconceptions about STD/HIV are very common. STD/HIV-related knowledge, perceived susceptibility of contracting HIV, and perceived efficacy of condom use for HIV/AIDS prevention were significantly associated with anticipated or actual risk behaviors during the infectious period. CONCLUSION: Intervention programs are in urgent need to break the cycle of STD infection in China. These would include promotion of relevant knowledge, condom use, and public education. PMID- 12897681 TI - Provision of syndromic treatment of sexually transmitted infections by community pharmacists: a potentially underutilized HIV prevention strategy. AB - BACKGROUND: Sexually transmitted infections (STIs) are known risk factors for HIV infection. GOAL: The goal of this study was to assess the current and potential future role that community pharmacists in Western Cape, South Africa play in the treatment of STIs. STUDY DESIGN: A cross-sectional survey of community pharmacists in the Western Cape region of South Africa. A face-to-face interview that ascertained experience with requests from patients for STI treatment, current STI treatment practices, and willingness to provide syndromic STI treatment was administered to head pharmacists. RESULTS: Ninety pharmacies were selected and 85 (94%) of the head pharmacists participated; 55 from an urban area and 30 from a rural area. Pharmacists reported a median of 40 urban clients and 25 rural clients who sought STI treatment from community pharmacists. When provided with a hypothetical clinical situation, 13% of urban and 17% of rural pharmacists identified the correct medication for male urethral discharge, 8% of urban pharmacists and none of the rural pharmacists identified correct treatment for genital ulcers, and none of the pharmacists identified the correct medication for vaginal discharge. Fifty-three percent of pharmacists in urban regions and 47% of pharmacists in rural regions expressed willingness to provide syndromic STI treatment. Independent predictors of willingness to provide syndromic treatment were knowledge of the link between HIV transmission and STIs (adjusted odds ratio [OR]: 13.78; 95% CI: 2.69, 70.66), past experience prescribing syndromic STI treatment (OR: 11.1; 95% CI: 1.14, 108.6), and male gender (OR: 4.38; 95% CI: 1.15, 16.7). CONCLUSIONS: Pharmacists are frequently called upon to provide STI treatment but have limited knowledge of correct treatment recommendations. Training pharmacists to provide syndromic STI treatment may be one strategy to reduce STI morbidity and HIV transmission. PMID- 12897682 TI - STD program activity in state policy processes, 1995 and 2000. AB - BACKGROUND: Generating state policy support for sexually transmitted disease (STD) prevention is crucial; however, little is known about policy activity by STD programs. GOALS: The goals of the study were to identify state-level policy behaviors by state STD programs and to examine the association between policy behavior and selected state characteristics. STUDY DESIGN: Information was gathered through a survey of STD directors in 2001. Policy activities were recalled for 1995 and 2000 and compared with several state characteristics. RESULTS: A majority of state STD programs reported at least one policy behavior in the state policy process for 1995 and for 2000. An increase was observed with the provision of state legislative testimony, participation in STD coalitions, and dissemination of information to policy-makers by STD programs. CONCLUSION: Reported policy activity may be evidence of an emerging environment enabling state STD directors to engage in public policy activity. An opportunity exists to improve state and local support for STD prevention. PMID- 12897683 TI - Behavioral surveillance surveys of the male clients of female sex workers in Hong Kong: results of three population-based surveys. AB - BACKGROUND: Male clients of female sex workers (FSWs) are an important group vulnerable to HIV infection. GOALS: The goal was to administer and assess behavioral surveillance surveys of male clients of FSWs in Hong Kong. STUDY DESIGN: Three population-based telephone surveys of males aged 18 to 60 were conducted from 1998 to 2001 (n=5159). RESULTS: Of all respondents, about 10.8% to 14% had patronized FSWs in the previous 6 months. Among male clients of FSWs, about 25% did not always use condoms, 4% to 6% had contracted a sexually transmitted disease, 16% had undergone HIV antibody testing, and 20% or more reported having four or more sex partners in the previous 6 months. There was no improvement in condom use over time. Cross-border (from Hong Kong to mainland China) commercial sex was common. CONCLUSION: The level of risk behaviors associated with male clients of FSWs remained substantial. The overall effectiveness and coverage of relevant prevention programs were questioned. PMID- 12897684 TI - Detection of human papillomavirus DNA on the external genitalia of healthy men and male patients with urethritis. AB - BACKGROUND: Only a few studies have been done involving detection of human papillomavirus (HPV) DNA on the external genitalia of men without genital warts, although many have been done for women. We conducted HPV DNA detection among healthy male volunteers and men with urethritis, both having no visible lesions on their external genitalia. GOAL: The goal of the study was to determine the detection rate of HPV DNA in volunteers and patients with urethritis and to determine risk factor(s) for positive DNA. STUDY DESIGN: This was a prospective clinical study. RESULTS: HPV DNA was found in 1.3% of 75 volunteers and in 18.5% of 130 patients with urethritis. DNA of a high-intermediate oncogenic risk was more predominant than the low-risk type. Among various risk factors, only a history of STD was a significant factor for the positive detection of HPV DNA in multiple regression analysis. CONCLUSION: HPV DNA was found in patients with urethritis more frequently than in volunteers, probably because the former had higher sexual activity. PMID- 12897685 TI - Adjudication history and African American adolescents' risk for acquiring sexually transmitted diseases: an exploratory analysis. AB - BACKGROUND: Little is known about sexually transmitted disease (STD) risk behavior among adjudicated African American adolescents. GOAL: The goal of the study was to compare STD-associated risk profiles of African American adolescents reporting a history of adjudication and those not reporting adjudication. STUDY DESIGN: A cross-sectional survey of 304 African American adolescent males and females (aged 15-21 years) was conducted. Adolescents were recruited from primary care clinics and through outreach activities. RESULTS: Twenty-six percent of the adolescents reported adjudication. After adjusting for gender, adjudicated adolescents were about 3.6 and 4.5 times, respectively, more likely than nonadjudicated adolescents to report ever having one of three STDs (gonorrhea, chlamydia, and trichomoniasis) or to report having one of these in the past 90 days. Reporting recent sex with someone known or suspected of having an STD was about nine times more likely among adjudicated adolescents, and they were about 2.6 times more likely than their nonadjudicated counterparts to report using drugs or alcohol during their last sexual experience and 2.2 times more likely to report frequent sex in the past 90 days. CONCLUSIONS: African American adolescents with a history of adjudication may have greater risk for acquisition of STDs than their peers not reporting adjudication. PMID- 12897686 TI - Human papillomavirus infection in men who have sex with men participating in a Dutch gay-cohort study. AB - BACKGROUND: To develop strategies for prevention and early treatment of human papillomavirus (HPV) anal and penile cancer, a better understanding of related sexual behavior risk factors is needed. GOAL: The goal of this study was to establish the prevalence of anal and coronal sulcus HPV in a group of men who have sex with men participating in a Dutch gay-cohort study, to identify risk factors associated with HPV infection in this group, and to investigate the presence of identical HPV types in couples with stable relationships. STUDY DESIGN: A cross-sectional study of 241 HIV-negative and 17 HIV-positive men who have sex with men visiting the sexually transmitted disease clinic of the Erasmus MC for a regular and scheduled examination. Participants underwent a routine venereological examination including HIV serologic analysis, and swabs were taken from the coronal sulcus and anus for HPV DNA testing. All subjects were asked to complete a questionnaire on sexual risk behavior. RESULTS: HPV DNA was detected at the coronal sulcus in 23.5% of the HIV-positive men and in 15.8% of the HIV negative men (P=0.492). In anal specimens, HPV DNA was detected in 64.7% of the HIV-positive men and 32.8% of the HIV-negative men (P=0.015). High-risk HPV types (P=0.007) and 2 or more different HPV genotypes (P=0.006) were seen more often in anal specimens of HIV-positive persons than in specimens of HIV-negative persons. A factor possibly associated with the presence of anal HPV infection was a concomitant anal infection with Chlamydia trachomatis, gonococci, or herpes simplex virus (P=0.059). In only 16.7% of HPV-positive steady couples, both companions showed the presence of one or more identical HPV genotypes. CONCLUSION: In this study, anal HPV DNA was detected more often than HPV DNA at the coronal sulcus. HIV positivity was associated with a higher prevalence of high-risk, but not with low-risk HPV types, at the anus. No association was found between HIV positivity and presence of high-risk HPV at the coronal sulcus. No sexual behavioral determinants for the presence of HPV could be identified. Concomitant anal infection with C trachomatis, gonococci, or herpes simplex virus may be associated with HPV infection. In the majority of steady couples, partners were infected with different HPV types. PMID- 12897687 TI - Bacterial vaginosis in pregnancy: diagnosis and treatment practices of physicians in San Diego, California, 1999. AB - BACKGROUND: Treating symptomatic bacterial vaginosis (BV) early in pregnancy may decrease preterm birth (PTB). Understanding how physicians manage BV is important for the development of interventions. GOAL: The goal was to determine the extent of knowledge and behaviors of physicians related to the diagnosis, treatment, and medical effects of BV in pregnant and nonpregnant patients. STUDY DESIGN: This was a cross-sectional survey. RESULTS: The study group consisted of 208 physicians who provided gynecologic care, including 102 (49%) who provided care to pregnant patients. Only 65% believed that there was a strong causal association between BV and PTB. Physicians who believed that BV causes PTB were much more likely to optimally manage vaginal infections (43% versus 7%). Only 12% of physicians prescribed oral metronidazole or clindamycin during the first trimester of pregnancy to treat BV. CONCLUSION: Physicians should be aware of the relation between symptomatic BV and PTB, seek a specific diagnosis for symptoms of vaginitis, use standard criteria to diagnose BV, and treat BV with effective regimens early in pregnancy. PMID- 12897688 TI - The syphilis reactor grid: help or hindrance for syphilis surveillance? AB - BACKGROUND: Health departments use reactor grids (sex, age, and serologic test for syphilis [STS] titer criteria) to determine which persons to evaluate for untreated syphilis. GOAL: The goal of the study was to assess reactor grid performance in Chicago and reactor grid use nationally in 1999 to 2000. STUDY DESIGN: We reviewed Chicago health department records to identify characteristics of persons with a reactive STS excluded from evaluation by reactor grid criteria and syphilis cases not meeting evaluation criteria. We surveyed health departments regarding reactor grid use. RESULTS: Of persons with a reactive STS, 46% did not meet criteria for health department evaluation, including 62% of men, 29% of women, and 21% with titers > or =1:8. The reactor grid would have excluded 17% of primary syphilis cases. Overall, 82% of health departments use reactor grids. CONCLUSIONS: Reactor grids are widely used and may exclude persons with infectious syphilis from health department evaluation, especially men. The impact of reactor grid use on syphilis control and surveillance in the United States should be evaluated. PMID- 12897689 TI - Community-based screening and treatment for STDs: results from a mobile clinic initiative. AB - BACKGROUND: The persistence of syphilis and other bacterial sexually transmitted diseases (STDs) in many areas of the United States suggests that innovative approaches to controlling these diseases are needed. GOAL: To evaluate the feasibility, acceptability, and yield of mobile community-based STD screening and treatment services in high STD incidence areas. STUDY DESIGN: Free, voluntary, confidential screening and treatment for STDs were conducted in high STD incidence neighborhoods of Baton Rouge, Louisiana, using a 32-foot mobile van. Demographic and behavioral data were obtained from participants. Participants were screened for syphilis, chlamydia, and gonorrhea and were also offered HIV testing. Community attitudes toward the screening program were assessed with street-intercept surveys conducted two weeks after screening events. RESULTS: From March 1997 to August 2000, 256 community-based screening events were held. During this period, 3110 blood samples were collected for syphilis testing, of which 37 (1.2%) new cases of syphilis were identified. Of the 2807 blood samples collected for HIV testing, 70 (2.5%) were positive. Of 2229 urine samples, 185 (8.3%) tested positive for Chlamydia trachomatis and 108 (4.9%) positive for Neisseria gonorrhoeae. Of 389 street-intercept surveys, 97% of respondents thought that neighborhood STD testing was a "good" or "very good" idea. CONCLUSION: Mobile community-based STD screening and treatment are feasible, identify high positivity of STDs, and are accepted by the community as an innovative approach to STD control. PMID- 12897690 TI - The role of STD outreach testing in times of dwindling STD prevention resources. PMID- 12897691 TI - Prevalence of sexually transmitted diseases among Latino immigrant day laborers in an urban setting--San Francisco. AB - BACKGROUND: Although rural Latino migrant laborers have been identified as a group at-risk for acquiring HIV, few studies have examined transmitted diseases (STDs) in urban, Latino migrant laborers. GOAL: To assess the prevalence of STDs in a sample of urban, migrant day laborers in San Francisco. STUDY DESIGN: A convenience sample of participants in the Day Laborer Project of the San Francisco Department of Public Health was screened for STDs from September 1994 to January 2001. Screening included serologic tests for syphilis and nucleic acid amplification tests for gonorrhea and chlamydia. RESULTS: A total of 292 clients participated in the screening program. All participants were male, Latino, and recent immigrants. Of the 235 persons screened for syphilis, secondary syphilis was diagnosed in one (0.4%) participant. Of the 198 persons screened for gonorrhea and chlamydia, 1 (0.5%) had gonorrhea and 7 (3.5%) had chlamydia. CONCLUSION: Urban Latino migrant day laborers are a population at-risk for infection with STDs. Community-based STD screening programs might be an effective way to detect STDs in this population. PMID- 12897692 TI - Participatory mapping of sex trade and enumeration of sex workers using capture recapture methodology in Diego-Suarez, Madagascar. AB - BACKGROUND: Capture-recapture methodology has been employed to estimate the size of hidden or difficult-to-reach human populations such as sex workers, homeless persons, and intravenous drug users. This study took place in the context of efforts to improve the quality of curative and preventive services for sexually transmitted infections offered to sex workers in Diego-Suarez, Madagascar in partnership with sex workers. GOAL: The goal of this study was to better understand sex-trading practices, map out sex work sites, and estimate the size of a mobile sex-worker population in Diego-Suarez, Madagascar using methods that can be reproduced in other resource-poor settings. STUDY DESIGN: Forty sex workers were trained to function as field researchers. Key informants in designated neighborhoods were identified and interviewed. Basic maps of sex-trade sites were designed by hand. The number of sex workers in Diego-Suarez was estimated using capture-recapture methodology. An educational brochure was used to "capture" sex workers during a first phase, and "recapture" 2 weeks later. RESULTS: Most sex trade was street based and mobile. Capture-recapture methods estimated a total of 2684 sex workers (95% CI: 2588-2780) in Diego-Suarez, or approximately 12% of the 15- to 49-year-old female population in the city. CONCLUSION: Geographic mapping and capture-recapture proved to be low-cost and relatively easy to use techniques for counting a hard-to-reach population, such as female street-based sex workers. Sex workers can be capable fieldworkers, and their active participation in research concerning sex work should be considered whenever possible. A high proportion of women of reproductive age were found to engage in sex work, indicating that there is an urgent need to scale up sexually transmitted infection and HIV prevention activities targeting sex workers, their sexual partners, and the general population. PMID- 12897693 TI - The personal cost of caring. PMID- 12897694 TI - Professionals' views and experiences of using outcome measures in palliative care. AB - In palliative care, outcome measures are increasingly used to aid clinical practice, conduct audit and research. The objective of this study was to elicit professionals' views and experiences of using outcome measures, paying special attention to the Palliative care Outcome Scale (POS). This article presents the results of a qualitative study of 26 professionals, experienced in using the POS, who were invited to participate in semi-structured telephone interviews. Of those invited, 22 people took part. Participants' comments were noted verbatim through the interviews and data subjected to content analysis. Analysis of data identified a number of key themes surrounding outcome measures, notably their reasons for use, application in clinical settings and a range of professionals' attitudes. The article concludes that understanding the process of outcome measures is important for improving their implementation. When undertaking further research, attention should be paid to the wider social, cultural and structural contexts, as factors that can influence the implementation of outcome measures. As the drive towards outcome measures continues, it is essential that measures are not developed in a vacuum. Instead they should always be informed by the needs and experiences of individuals and services. PMID- 12897695 TI - Application of the doctrine of double effect in end stage disease. AB - The aim of this article is to explore the doctrine of double effect and to identify whether it is appropriate to use the doctrine, and if so when, in end stage disease. The legal principles of the doctrine of double effect will be explored through the examination of three recent cases involving the death of a patient in the UK. The legal distinction between the doctrine of double effect and the intentional act of homicide will be discussed. PMID- 12897696 TI - The last 48 hours of life: a case study of symptom control for a patient taking a Buddhist approach to dying. AB - Caring for a patient dying of cancer can, at times, be extremely difficult. Sarah was 39 years old when she died, survived by her husband and two children aged 6 and 4 years. During the weeks leading up to her death, Sarah held extensive discussions with family and the multidisciplinary team. Her goal was to live the remainder of her last few hours as comfortable as possible and to die a "peaceful death". Terminal care is an important phase of life, one in which individuals have the right to expect quality of care to ensure that their death occurs with dignity. Sarah practised Buddhism on a daily basis. Issues of spirituality, serenity and peacefulness in dying were very important to her to ensure a good rebirth. PMID- 12897697 TI - The first 2 years of a palliative care consultation team in the Netherlands. AB - In January 1999 an integrated multidisciplinary palliative care consultation (PCC) team was established in Maastricht in the Netherlands. The team included experts in palliative care who had extensive experience in a variety of settings. One of the major tasks of the PCC team was to give support, information and advice to healthcare professionals caring for terminally ill patients. The PCC team was asked by the government to consecutively register and evaluate all consultations. This article describes these consultations, including information on the requesting caregivers, the patients, the questions asked and the recommendations given in the first 26 months. The results show that the PCC team served the needs of professional caregivers in a variety of settings. Most consultations concerned physical and pharmacological problems and the majority of recommendations were evaluated as positive. PMID- 12897698 TI - Dying at home: a care pathway for the last days of life in a community setting. AB - The Cancer Nursing Strategy (Department of Health (DoH), 2000a) expects nurses at senior level to lead the way in improving the organization and delivery of cancer care. Lack of forward planning can be a feature of community palliative care, and may lead to inappropriate crisis admission in the last days despite patients and relatives expressing a wish to stay at home. This article attempt to demonstrate how, though collaboration and effective team working, issues and/or problems encountered in the last days of life were managed by developing and promoting a care pathway for the last days of life. Although based on Ellershaw's pioneering work, this pathway was developed specifically for use in the community to reflect and address the particular problems encountered in this setting when caring for dying patients. PMID- 12897707 TI - Angiogenic and angiostatic factors in the molecular control of angiogenesis. AB - The vascular system that ensures an adequate blood flow is required to provide the cells with sufficient supply of nutrients and oxygen. Two different mechanisms of the formation of new vessels can be distinguished: vasculogenesis, the formation of the first primitive vascular plexus de novo and angiogenesis, the formation of new vessels from preexisting ones. Both processes are regulated by a delicate balance of pro- and anti-angiogenic factors. Physiologically, angiostatic mediators outweigh the angiogenic molecules and angiogenesis does not occur. Under certain conditions such as tumor formation or wound healing, the positive regulators of angiogenesis predominate and the endothelium becomes activated. Angiogenesis is initiated by vasodilatation and an increased permeability. After destabilization of the vessel wall, endothelial cells proliferate, migrate and form a tube, which is finally stabilized by pericytes and smooth muscle cells. Numerous soluble growth factors and inhibitors, cytokines and proteases as well as extracellular matrix proteins and adhesion molecules strictly control this multi-step process. The properties and interactions of angiogenic molecules such as VEGFs, FGFs, angiopoietins, PDGF, angiogenin, angiotropin, HGF, CXC chemokines with ELR motif, PECAM-1, integrins and VE-cadherin as well as angiostatic key players such as angiostatin, endostatin, thrombospondin, CXC chemokines without ELR motif, PEDF are discussed in this review with respect to their molecular impact on angiogenesis. PMID- 12897708 TI - Imaging the effects of anti-angiogenic treatments. AB - It is clear that various imaging modalities have given keen insight into the molecular mechanisms involved in anti-angiogenic treatments. A key to the advancement of anti-angiogenic therapy is not only the discovery of new drugs and treatments, but the analysis of the specific modes of action of these compounds in order to produce the next generation with greater effectiveness. While existing clinical methods incorporate the analysis of serum and urine to measure angiogenic factors, an imaging technique monitoring the effectiveness of anti angiogenic therapy would be a convenient, noninvasive, cost effective technique to aid in treatment planning and disease management. PMID- 12897709 TI - CT imaging of angiogenesis. AB - Tumor angiogenesis has significant implications in the diagnosis and treatment of various solid tumors. With the advent of fast, multi-slice CT scanners, CT imaging techniques capable of qualitative and quantitative analysis of tumor angiogenesis have been the subject of extensive investigation in the past 2 decades. The fundamental bases for CT imaging of angiogenesis are both the transport by blood flow of intravenously administered iodinated contrast material to tissue and the exchange by diffusion of these contrast molecules between the intravascular space and the extravascular interstitial space. With current fast CT scanners both tissue and vascular enhancement can be measured and traced over time at small time intervals to allow detailed modeling of the distribution of contrast agent in tissue. Both compartmental and distributed parameter models for contrast transport and exchange have been developed to quantify from the CT data the following angiogenesis related parameters: tissue blood flow, blood volume, mean transit time, contrast arrival time, capillary permeability surface area product and hepatic arterial fraction in case of the liver. This review addresses the following aspects of CT imaging of angiogenesis: 1) basic concepts related to the understanding of both compartmental and distributed parameter models; 2) comparison between both types of models; 3) practical issues with respect to the measurement of the arterial input function, which is required for the solution of both types of models; and, 4) illustration of the application of a distributed parameter model, the Johnson and Wilson model, in a number of experimental studies. PMID- 12897710 TI - Radiotracer-based strategies to image angiogenesis. AB - Tumour-induced angiogenesis plays an important role in tumour progression. Great efforts are made to develop therapeutic strategies to interfere with this process resulting in the starvation of the tumour. However, strategies to monitor conventional therapies seems to be inappropriate to control these approaches. Thus, there is a keen interest in developing methods supplying information about the corresponding therapeutical effects. Several radiotracer-based approaches focused on different targets in the angiogenic process are currently investigated. One class of tracers is based on matrix metalloproteinases inhibitors. These compounds show promising results in in vitro assays. However, initial data from in vivo studies using murine tumour models could not confirm successful non-invasive monitoring of MMP activity yet. Another strategy uses a radiolabelled single chain fragment against the ED-B domain of fibronectin, an extracellular matrix protein. Promising results demonstrated selective accumulation of the tracer in the tumour vasculature of a murine tumour model. Most of the studies are concentrated on the development of radiolabelled antagonists of the integrin alpha(v)beta(3). This heterodimeric transmembrane glycoprotein is involved in the migration of activated endothelial cells during formation of new vessels. Different compounds have been labelled with (18F), (111)In, (99m)Tc, (90)Y and several iodine isotopes. In in vitro assays most of them revealed high alpha(v)beta(3) affinity and selectivity. Moreover, in different murine tumour models successful non-invasive determination of alpha(v)beta(3) expression has been shown. Some of these approaches indicate that tumour-induced angiogenesis can be monitored in animal studies. Nevertheless, translation of these approaches into clinical settings allowing visualisation of tumour-induced angiogenesis in patients needs still to be demonstrated. PMID- 12897711 TI - Imaging matrix metalloproteinase expression in tumors. AB - metastatic Matrix metalloproteinases (MMPs) are zinc-dependent secreted or transmembrane enzymes constituting a family of over 21 proteolytic members that are capable of selectively digesting a wide spectrum of both extracellular matrix (ECM) and nonmatrix proteins. MMPs play a critical role in tumor growth, angiogenesis, and metastatic processes. MMP inhibitors (MMPIs) have been extensively investigated as anti-tumor drugs, although the clinical trials thus far have been disappointing. In order to better understand the role of MMPs in cancer growth and metastasis, as well as improve the therapeutic efficacy of MMPIs, there is a need to develop new procedures to assess and/or monitor MMP activity in vivo. In addition to determining whether MMPs are present in tumors, it would be desirable to have an imaging agent that better probes other processes associated with MMP overproduction, including angiogenesis and the establishment of the growth of metastatic lesions in distant organ sites. In this paper we review the studies relating to the recent development of in vivo imaging of MMP expression. One of the purposes of this review is to discuss the current status of imaging MMP expression, which includes the types of tracers being developed and the types of imaging modalities available. Although imaging MMP expression is a relatively new area of research, the progress thus far is highly promising. PMID- 12897712 TI - Radiopharmaceuticals for targeting the angiogenesis marker alpha(v)beta(3). AB - A common feature of solid tumors is the formation of new blood vessels (angiogenesis) within the tumor. A receptor called alpha(v)beta(3) is found on endothelial cells lining newly growing blood vessels at a higher density than on mature blood vessels. This receptor may provide a target for radioligands to permit imaging of a wide variety of solid tumors. The radioligands may range from macromolecules such as native ligands or monoclonal antibodies, to small proteins to very small peptides. The differing characteristics of these bio-molecules have an affect on target delivery and clearance time. PMID- 12897724 TI - Patterns of pollen cross-allergenicity. AB - Knowledge of patterns of pollen cross-reactivity is crucial for diagnostics and especially for formulation of immunotherapy vaccines in times of diminishing availability of pollen extract constituents. As phylogenetic relationships have become better clarified, it becomes apparent that cross-reactivity does reflect taxonomy in the very great majority of cases. Contradictory observations of unexpected cross-reactivity between unrelated plants, sometimes remarkably distant ones, require explanation. There are many proteins, presumably performing vital functions, that are tightly preserved throughout the evolutionary tree from plants to animals, such as profilins, lipid transfer proteins, and pathogenesis related proteins. These might function as panallergens. The small differences that exist between these ubiquitous proteins explain why these are frequently minor allergens not reacting in the majority of allergic sera. This review summarizes cross-reactivity studies with both crude pollen extracts and purified or recombinant allergenic proteins. The patterns of cross-allergenicity that emerge should be helpful in guiding both diagnostic and therapeutic decisions. PMID- 12897726 TI - Signaling molecules as therapeutic targets in allergic diseases. AB - A molecular understanding of physiologic and pathologic processes requires complete knowledge about the signal transduction mechanism of involved cells. Signal transduction research is a rapidly growing field in basic science. Unlike intercellular inflammatory mediators, signaling molecules show less functional redundancy. This allows inhibition of multiple cytokines/mediators by blocking one common signaling molecule. Interference with signaling pathways has shown significant potential for inhibition of fundamental processes as well as clinical phenotype of allergic diseases. The purpose of this review was to provide a theoretical classification of signaling molecules based on their function and to analyze various strategies for developing effective signaling inhibitors for allergic diseases. PMID- 12897728 TI - Allergic and nonallergic forms of atopic diseases. AB - Atopic dermatitis, allergic rhinitis, and asthma are atopic diseases that develop on a complex genetic background, the so-called atopic diathesis. Although they target different organs, in most patients they are characterized by the presence of elevated total serum IgE levels. However, a subgroup of atopic patients exhibits normal IgE levels and mechanisms contributing to the so-called "intrinsic" or "nonallergic form" have been the matter of intensive research work in the last years. Because of the rapid advancements in the research field of atopic diseases, it now becomes possible for the first time to delineate a new disease classification of allergic and nonallergic subtypes of atopic diseases, thereby bringing hope to the clinician for a more specific treatment approach for each subgroup of these patients. PMID- 12897729 TI - Pets--good or bad for individuals with atopic predisposition? PMID- 12897730 TI - Cockroach allergen abatement: the good, the bad, and the ugly. PMID- 12897731 TI - Asthma and pregnancy. PMID- 12897732 TI - Perimenstrual asthma: a syndrome without known cause or cure. AB - Perimenstrual worsening of asthma has been documented in 30% to 40% of asthmatic women. This increase in symptoms has been backed up by increased health care use perimenstrually, as well as by cyclic variation in peak expiratory flows. The cause of perimenstrual asthma (PMA) remains unclear. Fluctuations in hormone levels, their ratios, or both are a plausible explanation but have not been demonstrated with any consistency. Influences of sex hormones on inflammation is an area of future research, as are hormone-induced changes in smooth muscle function and beta-adrenergic receptors, prostaglandin levels, and fluid retention in the bronchial mucosa. In the light of the high prevalence of PMA, it is difficult to understand why there has been no randomized controlled trial of hormone therapy. Nevertheless, several case reports have suggested beneficial effects of estrogens, progestins, and their combination. In light of these positive case reports, well-designed, double-blind studies of sufficient sample size should now be performed to give treatment of PMA an evidence base. PMID- 12897733 TI - Asthma morbidity during pregnancy can be predicted by severity classification. AB - BACKGROUND: The 1993 National Asthma Education Program Working Group on Asthma and Pregnancy defined asthma severity as mild, moderate, or severe on the basis of symptoms and spirometry, but no studies have evaluated the relationship between this classification system and subsequent asthma morbidity during pregnancy. OBJECTIVE: The objective of this study was to evaluate the relationship between asthma severity classification during pregnancy and gestational asthma exacerbations. METHODS: Asthma severity was defined according to the 1993 classification, adjusted to include medication requirements, in a volunteer sample of 1739 pregnant asthmatic patients who were less than 26 weeks' gestation. RESULTS: Initial asthma classification (mild, moderate, or severe) was significantly related to subsequent asthma morbidity during pregnancy (hospitalizations, unscheduled visits, corticosteroid requirements, and asthma symptoms during labor and delivery). Exacerbations during pregnancy occurred in 12.6% of patients initially classified as mild, 25.7% of patients classified as moderate, and 51.9% of patients classified as severe (P <.001). Asthma morbidity was similar, whether patients were classified as moderate or severe by symptoms and spirometry or by medication requirement. Thirty percent of initially mild patients were reclassified as moderate-severe during pregnancy, and 23% of the initially moderate-severe patients were reclassified as mild later in pregnancy; asthma morbidity in these patients changed accordingly. CONCLUSION: The National Asthma Education Program Working Group on Asthma and Pregnancy classification of asthma severity, adapted to include medication use, predicts subsequent asthma morbidity during pregnancy. PMID- 12897734 TI - Pet-keeping in childhood and adult asthma and hay fever: European community respiratory health survey. AB - BACKGROUND: Whether pet-keeping early in life protects against or promotes allergy remains unclear. OBJECTIVE: Our aim was to examine the effects of childhood pet-keeping on adult allergic disease in a large international population-based study, including information on sensitization, adult pet keeping, and pet prevalence in the populations. METHODS: We used information from structured interviews (n = 18,530) and specific IgE to common aeroallergens in blood samples (n = 13,932) from participants in the European Community Respiratory Health Survey (ECRHS) to analyze the associations between keeping pets and adult asthma and hay fever. RESULTS: Keeping cats in childhood was associated with asthma only among atopic subjects, an association that varied between centers (P =.002) and was stronger where cats where less common (< 40% cats: odds ratio(wheeze) [OR(wheeze)] = 1.84, 95% CI = 1.31-2.57; 40%-60% cats: OR(wheeze) = 1.33, 95% CI = 1.10-1.61; > or =60% cats: OR(wheeze) = 0.98, 95% CI = 0.73-1.33). Dogs owned in childhood or adulthood were associated with asthma among nonatopic subjects (childhood: OR(wheeze) = 1.28, 95% CI = 1.13-1.46; adulthood: OR(wheeze) = 1.31, 95% CI = 1.14-1.51; both: OR(wheeze) = 1.69, 95% CI = 1.40-2.04). In atopic subjects, those who had owned dogs in childhood had less hay fever (OR = 0.85; 95% CI = 0.73-0.98) and no increased risk of asthma (OR(wheeze) = 1.01, 95% CI = 0.87-1.17). Respiratory symptoms were more common in subjects who had owned birds during childhood (OR(wheeze) = 1.12; 95% CI = 1.02 1.23) independent of sensitization. CONCLUSIONS: The effects of pet-keeping in childhood varied according to the type of pet, the allergic sensitization of the individual, and the wider environmental exposure to allergen. Cats owned in childhood were associated with more asthma in sensitized adults who grew up in areas with a low community prevalence of cats. Dogs owned in childhood seemed to protect against adult allergic disease but promote nonallergic asthma. PMID- 12897735 TI - Efficacy of local nasal immunotherapy for Dp2-induced airway inflammation in mice: Using Dp2 peptide and fungal immunomodulatory peptide. AB - BACKGROUND: Local nasal immunotherapy (LNIT) is an effective immunotherapy. Peptides derived from the group 2 allergen of Dermatophagoides pteronyssinus, Dp2 28-40 and Dp2 28-40A, and fungal immunomodulatory peptide (FIP) have been shown to act as T(H)1 potential and response-inducing adjuvant. LNIT by the use of Dp2 peptides in conjunction with FIP were investigated. OBJECTIVE: We sought to determine whether Dp2-induced airway inflammation in mice could be downregulated by Dp2 peptides or a mixture of Dp2 peptides with FIP. METHOD: Mice were sensitized with rDp2 followed by LNIT with Dp2 peptides, FIP, or FIP and a mixture of Dp2 peptides. After intratracheal challenge with rDp2, the airway inflammation and hyperresponsiveness were determined by bronchoalveolar lavage fluid (BALF) analysis and methacholine challenge. RESULTS: Both Dp2 peptides and FIP were able to inhibit rDp2-induced airway inflammation and airway hyperresponsiveness. An increase in IFN-gamma and a decrease in IL-5 in BALF and sera were found after LNIT with Dp2 peptides, FIP, and mixtures of both. Serum levels of TGF-beta were reduced after LNIT with FIP and Dp2 28-40. Penh values were significantly decreased after methacholine challenge in both the early and late phase. CONCLUSIONS: LNIT with allergen-derived peptides and FIP can produce an anti-inflammatory effect on allergen-induced airway inflammation. LNIT with selected peptides and FIP might be a good alternative therapy for allergic airway disease. PMID- 12897736 TI - Factors influencing symptom expression in children with bronchial hyperresponsiveness at 10 years of age. AB - OBJECTIVES: We sought to identify factors associated with wheezing symptoms in children found to have bronchial hyperresponsiveness (BHR) at 10 years of age. METHODS: Children were seen at birth, 1, 2, 4 and 10 years of age in an entire population birth cohort study (n = 1456). At each stage information was collected prospectively on genetic and environmental risk factors for BHR. Skin prick testing was performed at 4 and 10 years of age. Spirometry and methacholine bronchial challenge were conducted at 10 years of age when BHR was considered present if PC(20) FEV(1) was < 4.0 mg/mL. In children with BHR at 10 years of age, factors independently associated with current wheezing were determined by logistic regression. RESULTS: BHR was identified in 169 10-year-olds at bronchial challenge, 55.6% of whom manifested current wheeze. In children with BHR, current wheezers had higher Log(10) total IgE and greater BHR than those who had never wheezed. Symptomatic BHR was independently associated with atopic sensitization (P <.001) and maternal asthma (P =.011) at 10 years of age. If only factors present in the first 4 years of life were considered, parental smoking at 4 years of age (P =.021), maternal asthma (P =.017), and atopic sensitization at 4 years of age (P =.004) were independently associated with symptomatic BHR at 10 years of age. CONCLUSIONS: Symptomatic BHR is associated with greater degrees of BHR and higher total IgE. Heredity, atopy, and environmental exposure might influence symptom expression in children with BHR. PMID- 12897737 TI - Impulse oscillometry provides an effective measure of lung dysfunction in 4-year old children at risk for persistent asthma. AB - BACKGROUND: Objective lung function measurements are routinely used to diagnose and manage asthma, but their utility for young children has not been defined. OBJECTIVE: Bronchodilator responses were measured by means of impulse oscillometry (IOS) and compared with conventional spirometry to determine the value of lung function measures in 4-year-old asthma-prone children. METHODS: The study participants were in the Childhood Asthma Prevention Study (National Institute of Health/National Institute of Allergy and Infectious Diseases) and at risk for asthma. At age 4 years, concurrent asthma was determined by using a previously validated modified American Thoracic Society questionnaire. Children performed IOS and spirometry before and after albuterol administration and underwent skin prick testing to 13 common allergens to assess atopy. IOS measures were as follows: airways resistance at 5 Hz and 10 Hz, airways reactance at 5 Hz and 10 Hz, and resonant frequency. RESULTS: Asthmatic patients versus nonasthmatic patients significantly differed in their IOS-assessed bronchodilator responses through Delta resistance at 5 Hz (medians, 27% vs 17%; P =.02) and Delta resistance at 10 Hz (24% vs 16%; P =.03). Because atopic children who have frequent wheezing are at risk for persistent asthma, the data were analyzed in regard to atopic patients with or without asthma. IOS strongly distinguished atopic asthmatic children through Delta resistance at 5 Hz (36% vs 13%, P =.007), Delta resistance at 10 Hz (25% vs 11%, P =.02), and Delta reactance at 10 Hz (47% vs 12%, P =.03). Conventional spirometry did not establish similar statistically significant findings. CONCLUSION: IOS bronchodilator responses are remarkably abnormal in 4-year-old children, who are most likely to have persistent asthma. IOS is a useful diagnostic tool in early asthma development and might be a helpful objective outcome measure of early interventions. PMID- 12897738 TI - TLR4 gene variants modify endotoxin effects on asthma. AB - BACKGROUND: Environmental exposure to endotoxin might have a crucial role in immune maturation and development of asthma. OBJECTIVE: The aim of this study was to investigate whether the effect of endotoxin concentration in settled house dust on asthma is modified by the presence of variation in the TLR4 gene. METHODS: We performed a cross-sectional study within the German follow-up of the European Community Respiratory Health Survey. Multivariate logistic regression analysis and nonparametric effect estimates (S-Plus) were applied to examine the association between endotoxin exposure and diagnosed asthma, related clinical symptoms, and bronchial hyperreactivity (BHR) stratified for noncarriers and carriers of G299/I399 polymorphism in the TLR4 gene. RESULTS: In the noncarrier group (n = 279), the prevalence of asthma was significantly increased with elevated endotoxin levels in house dust with adjusted odds ratio 6.24 (95% CI, 1.33-29.17) in the second tertile, and 4.54 (95% CI, 0.94-21.96) in the third tertile compared with the lowest endotoxin tertile. The carriers of the polymorphisms (n = 55) showed a nonsignificant trend to have a lower risk of asthma (crude odds ratio, 0.67; 95% CI, 0.06-8.06 for the second tertile and 1.33; 95% CI, 0.17-10.58 for the third tertile). We found a similar association for wheeze and endotoxin exposure that was also attenuated in subjects with G299/I399 polymorphisms. CONCLUSIONS: The G299/I399 polymorphisms were associated with a modified response to endotoxin, but the functional relationship still needs clarification. PMID- 12897739 TI - The effects of an anti-CD11a mAb, efalizumab, on allergen-induced airway responses and airway inflammation in subjects with atopic asthma. AB - BACKGROUND: Efalizumab is a humanized IgG(1) mAb against the lymphocyte function antigen-1 (LFA-1) alpha chain, CD11a. Blocking of LFA-1/intercellular adhesion molecule interactions could inhibit asthmatic inflammation by blocking adhesion and activation of LFA-1-positive leukocytes. OBJECTIVE: A randomized, double blinded, placebo-controlled, parallel group, multicenter study investigated the effects of efalizumab on allergen-induced airway responsiveness and airway inflammation. METHODS: Thirty-five nonsmoking subjects with mild allergic asthma were randomized to receive efalizumab (n = 24) or placebo (n = 11) in 8 weekly subcutaneous doses (0.7 mg/kg conditioning dose followed by 7 weekly doses of 2.0 mg/kg). Allergen challenges were performed at screening and after 4 and 8 weeks of treatment. Samples of sputum (n = 18 subjects) and blood (n = 35 subjects) were collected the day before challenges, and sputum was collected again at 7 and 24 hours after each challenge. Nonparametric tests were used to compare allergen induced differences between efalizumab and placebo groups. RESULTS: Subjects receiving efalizumab developed headache (48%) and flu syndrome (28%) compared to subjects receiving placebo (0%). After 8 weeks of efalizumab, the maximum late percent fall in FEV(1) (late asthmatic response) was inhibited by 50%, but neither the late response nor the late area under the curve was statistically different than placebo (P =.098 and.062, respectively). Efalizumab had no effect on the maximum early percent fall in FEV(1) (early asthmatic response) or early area under the curve compared to placebo (P >.59). Efalizu-mab significantly reduced the postallergen increase in sputum EG2-positive cells and metachromatic cells (P <.05). No other comparisons were statistically different. CONCLUSIONS: Blocking of LFA-1/intercellular adhesion module interactions by efalizumab inhibits the development of allergen-induced cellular inflammatory responses measured in induced sputum and might attenuate the late asthmatic response. Larger studies are needed to confirm this. PMID- 12897740 TI - Abatement of cockroach allergen (Bla g 1) in low-income, urban housing: A randomized controlled trial. AB - BACKGROUND: Clinically relevant reductions in exposure to cockroach allergen, an important risk factor for asthma in inner-city households, have proven difficult to achieve in intervention trials. OBJECTIVE: This study investigated a method for the abatement of cockroach allergen in low-income, urban homes. The goal was to reduce mean Bla g 1 concentrations below the previously proposed thresholds for allergic sensitization and asthma morbidity. METHODS: A prerandomized, nonmasked trial with 16 intervention and 15 control homes was conducted. Study inclusion was based on 50 to 500 cockroaches trapped in a 3-day period. The interventions consisted of occupant education, placement of insecticide bait, and professional cleaning. Vacuumed dust and multiple swab samples were collected at 0, 1, 2, 4, and 6 months in intervention homes and at 0 and 6 months in control homes. Room maps containing cockroach and allergen data were used to guide and monitor the interventions. RESULTS: From 0 to 6 months among intervention homes, geometric mean Bla g 1 concentrations (U/g dust) decreased from 633 to 24 on kitchen floors (96% reduction), from 25 to 4.3 on living room floors/sofas (83% reduction), from 46 to 7.3 on bedroom floors (84% reduction), and from 6.1 to 1.0 in bedroom beds (84% reduction). These reductions, with the exception of that on the bedroom floor (P =.06), were statistically significant relative to changes in control homes. CONCLUSIONS: Substantial reductions in cockroach allergen levels can be achieved in inner-city homes. In this study, allergen levels were reduced below the sensitization threshold (2 U/g) in beds, arguably the most relevant site for exposure, and below the asthma morbidity threshold (8 U/g) on bedroom floors and living room floors/sofas. The level on kitchen floors, although reduced 96%, remained above the asthma morbidity threshold. Future studies will test the intervention's effectiveness in asthma prevention trials. PMID- 12897741 TI - The prevalence of rat allergen in inner-city homes and its relationship to sensitization and asthma morbidity. AB - BACKGROUND: Rat allergen has proved to be an important cause of IgE-mediated hypersensitivity in the occupational setting. The prevalence and significance of rat allergen in homes has not been studied. OBJECTIVE: The purpose of this study was to determine the prevalence of rat allergen in the homes of inner-city children with asthma and to examine the relationship between rat allergen exposure, sensitization, and asthma morbidity. METHODS: We developed a new monoclonal-based ELISA to determine the prevalence of rat allergen in dust samples from inner-city homes of the National Cooperative Inner-City Asthma Study population. Home characteristics were evaluated to detect variables that were associated with the presence of rat allergen. Data were also analyzed to assess the relationship between the presence of rat allergen, sensitization, and asthma morbidity. RESULTS: Thirty-three percent of inner-city homes had detectable rat allergen (Rat n 1). The presence of rat allergen was associated with reported rat and mouse infestation, as well as evidence of mouse infestation on home inspection. Twenty-one percent of the participants were sensitized to rat allergen; however, sensitization was not more common when rat allergen was found in the home. The number of hospitalizations, unscheduled medical visits, and days with slowed activity because of asthma were significantly increased in those individuals who were both sensitized and exposed to rat allergen. CONCLUSIONS: Rat allergen sensitization and exposure are associated with increased asthma morbidity in inner-city children. PMID- 12897742 TI - Effect of inhaled endotoxin on airway and circulating inflammatory cell phagocytosis and CD11b expression in atopic asthmatic subjects. AB - BACKGROUND: In a cohort of 8 normal and 10 allergic asthmatic volunteers, we previously reported that inhalation of 5 microg of endotoxin (LPS) induced airway inflammation that correlated with CD14 expression that was, in turn, correlated with eosinophil numbers in the airway. Macrophage and neutrophil functions have been reported to be modified by endotoxin in vitro and in vivo, and response to endotoxin is mediated largely by airway phagocytes and related to allergic inflammation. OBJECTIVE: We sought to examine functional and cell-surface phenotype changes in phagocytes recovered from atopic asthmatic subjects after endotoxin challenge. METHODS: Sputum and peripheral blood from 10 allergic asthmatic subjects was recovered after saline and LPS challenge. Assessment of phagocytosis and cell-surface phenotype (CD11b, CD14, and CD64) was performed on phagocytes obtained from sputum (n = 7) and blood samples (n = 10). RESULTS: Phagocytosis of blood and sputum phagocytes was blunted after LPS challenge in a fashion that correlated with the increase in airway neutrophils after LPS challenge. Cell-surface expression of CD14 (membrane-bound CD14) was increased in sputum cells, whereas CD11b was decreased in sputum and circulating phagocytes. Baseline expression of CD11b in blood correlated with the magnitude of the neutrophil response after LPS inhalation, as well as (inversely) with baseline airway eosinophil levels. CONCLUSIONS: Inhalation of endotoxin at levels adequate to induce a neutrophil influx to the airways (but not systemic symptoms) results in decreased phagocytosis in both airway and circulating cells and modifies CD11b expression in a way that implicates its involvement in phagocyte responsiveness to inhaled LPS. PMID- 12897743 TI - Exposure and sensitization to indoor allergens: association with lung function, bronchial reactivity, and exhaled nitric oxide measures in asthma. AB - BACKGROUND: Exposure to high levels of allergens in sensitized asthmatic patients causes worsening of pulmonary function in experimental studies. Chronic exposure to lower, naturally occurring levels of allergens might increase the severity of asthma. OBJECTIVE: We sought to study the associations between sensitization and exposure to common indoor allergens (dust mite, cat, and dog) in the home on pulmonary function, exhaled nitric oxide (eNO), and airway reactivity in asthmatic patients. METHODS: Dust samples were collected from the living room carpet and mattress of 311 subject's homes, and Der p 1, Fel d 1, and Can f 1 concentrations were measured by using ELISAs. Spirometry, nonspecific bronchial reactivity, and eNO were measured. RESULTS: Subjects both sensitized and exposed to high levels of sensitizing allergen had significantly lower FEV(1) percent predicted values (mean, 83.7% vs 89.3%; mean difference, 5.6%; 95% CI, 0.6% 10.6%; P =.03), higher eNO values (geometric mean [GM], 12.8 vs 8.7 ppb; GM ratio, 0.7; 95% CI, 0.5-0.8; P =.001), and more severe airways reactivity (PD(20) GM, 0.25 vs 0.73 mg; GM ratio, 2.9; 95% CI, 1.6-5.0; P <.001) compared with subjects not sensitized and exposed. No significant effect of the interaction between sensitization and exposure was found for FEV(1) percent predicted and eNO values. However, there was a significant effect of the interaction between sensitization and exposure to any allergen (P =.05) and between sensitization and exposure to cat allergen (P =.04) for nonspecific bronchial reactivity. CONCLUSION: Asthmatic subjects who are exposed in their homes to allergens to which they are sensitized have a more severe form of the disease. PMID- 12897744 TI - Cysteinyl leukotrienes induce nuclear factor kappa b activation and RANTES production in a murine model of asthma. AB - BACKGROUND: It has been demonstrated that both cysteinyl leukotrienes (cysLTs) and cytokines are involved in the pathophysiology of bronchial asthma. Nonetheless, the exact mechanism involved in the interaction between these 2 molecules has yet to be determined. OBJECTIVE: The aim of the present study was to determine the effects of cysLTs on allergic airway inflammation and allergen specific cytokine production in a murine model of asthma. METHODS: Four groups of BALB/c mice (control mice, Dermatophagoides farinae allergen-sensitized mice, pranlukast cysLT receptor antagonist-treated allergen-sensitized mice, and dexamethasone-treated allergen-sensitized mice) were examined. RESULTS: Allergen sensitized mice exhibited increased airway responsiveness and inflammation. Pranlukast-treated mice showed significant attenuation of these changes concomitant with reduction of T(H)2 cytokine and IFN-gamma production by isolated lung mononuclear cells (MNCs). A much stronger inhibition of all cytokines was noted in dexamethasone-treated mice. Pranlukast also significantly inhibited production of RANTES and activation of nuclear factor kappa B (NF-kappa B) in the isolated lung MNCs. Leukotriene D(4) stimulated isolated lung MNCs to produce RANTES but not any other cytokines and also activated NF-kappa B in these cells. CONCLUSIONS: Our results suggest that cysLTs activate NF-kappa B and induce RANTES production from isolated lung MNCs, which in turn might cause migration of eosinophils and activated T lymphocytes into the airway. PMID- 12897745 TI - Endothelial protein C receptor-dependent inhibition of human eosinophil chemotaxis by protein C. AB - BACKGROUND: Eosinophil infiltration is a characteristic feature of allergic inflammation. Allergic responses are associated with local activation of the coagulation pathway and accumulation of fibrin. OBJECTIVE: We tested whether protein C and activated protein C (APC), which are endogenous anti-inflammatory coagulation inhibitors, affect eosinophil function. METHODS: Eosinophils were from venous blood of healthy donors. Cell migration and apoptosis were studied by using micropore filter assays and fluorometry, respectively. Receptor expression was investigated by means of RT-PCR and SDS-PAGE of immunoprecipitated protein. RESULTS: Protein C and APC had no significant chemotactic effects on eosinophils. Eosinophils pretreated with protein C or APC showed significantly reduced migration toward chemoattractants. No effect of either protein C preparation was seen in eosinophil apoptosis assays. The inhibiting effect on migration was reversed by an antibody against the endothelial protein C receptor (EPCR). Synthesis of EPCR by eosinophils is suggested by demonstration of receptor mRNA expression and detection of metabolically labeled receptor protein. CONCLUSIONS: Data suggest that an EPCR is expressed by eosinophils whose activation with protein C or APC arrests directed migration. Protein C-affected eosinophil chemotaxis is a novel thrombin-independent component of the protein C pathway. PMID- 12897746 TI - Associations between total serum IgE levels and the 6 potentially functional variants within the genes IL4, IL13, and IL4RA in German children: the German Multicenter Atopy Study. AB - BACKGROUND: Increased total serum IgE levels are a common characteristic of atopic disorders. Six potentially functional variants, including C-590T in the IL4 gene, C-1055T and Arg130Gln in the IL13 gene, and Ile50Val, Ser478Pro, and Gln551Arg in the IL4RA gene, have been evaluated for their involvement in the control of total serum IgE levels and related atopic disorders, but the results of these studies have been inconsistent. OBJECTIVE: We examined whether these 6 variants had genotypic effects on total serum IgE levels in 823 unrelated German children from a large infant cohort, the German Multicenter Atopy Study. METHODS: Marginal effect models were used for the analyses of the repeated IgE measurements. Weighted linear regression and family-based tests of association were performed to minimize the possibility of spurious effects caused by selection bias or confounding on the basis of ethnic background. RESULTS: There are significant associations between increased total serum IgE levels and 2 variants in the IL13 gene (P <.005 and.0002 for Arg130Gln and C-1055T, respectively). These genetic effects are unlikely to be due to solely linkage disequilibrium between 2 polymorphisms, population stratification, or nonrepresentative samples. In addition, exposure to maternal smoking appears to modify the above effects on total serum IgE levels. However, no statistical association was observed between this quantitative phenotype and the other 4 variants examined. CONCLUSION: These findings suggest that variants C-1055T and Arg130Gln of the IL13 gene might play an important role on total serum IgE production in this study population. PMID- 12897747 TI - Exposure to endotoxin and allergen in early life and its effect on allergen sensitization in mice. AB - BACKGROUND: Exposure to endotoxins, allergens, or both in early life might regulate the development of tolerance to allergens later in life. OBJECTIVE: We investigated whether continuous exposure of infant mice to aerosolized endotoxin, allergen, or both inhibits subsequent allergen-induced immune and inflammatory responses. METHODS: Infant BALB/c mice were pre-exposed to aerosolized endotoxin, ovalbumin (OVA), or both (3 times a week for the first 4 weeks of life) before systemic sensitization (days 1-14) and repeated airway challenge (days 28-30) with OVA. RESULTS: Compared with that seen in negative control animals, systemic sensitization and airway allergen challenges induced high serum levels of allergen-specific IgE (0.7 +/- 0.09 vs 0.02 +/- 0.01 OD units), predominant T(H)2 type cytokine production (IL-5 by splenic mononuclear cells in vitro, 1.2 +/- 0.2 vs 0.04 +/- 0.06 ng/mL), airway inflammation (bronchoalveolar lavage fluid leukocytes, 125 +/- 15 vs 64 +/- 7/microL; eosinophils, 28 +/- 5 vs 1 +/- 0/microL) and development of in vivo airway hyperreactivity (maximal enhanced pause, 11 +/- 1.9 vs 4 +/- 0.2). Pre-exposure with LPS before sensitization increased production of specific IgG2a (67 +/- 10 vs 32 +/- 5 U/mL) but failed to prevent T(H)2-mediated immune responses. Pre-exposure with OVA or with OVA plus LPS completely suppressed allergen sensitization, airway inflammation, and development of in vivo airway hyperreactivity; values were similar to those of negative control animals. Inhibition was due to allergen-specific T-cell anergy indicated by omitted allergen-specific T(H)2 and T(H)1 immune responses. In addition, combined exposure to endotoxin and allergen induced a general shift toward an unspecific T(H)1 immune response. CONCLUSION: Exposure with endotoxins before allergen sensitization is not able to induce unresponsiveness but might decrease the susceptibility for sensitization to a variety of common allergens. PMID- 12897748 TI - Complement activation in a model of chronic fatigue syndrome. AB - BACKGROUND: A need exists to identify biological markers in chronic fatigue syndrome (CFS). OBJECTIVE: To use an exercise and/or allergen challenge to induce the symptoms of CFS and to identify a biological marker that correlates with these symptoms. METHODS: Patients with CFS (n = 32) and age-matched, normal control patients (n = 29) exercised for 20 minutes on a stationary bike at 70% of their predicted max work load (Watts). Patients from each group with positive skin test results were also challenged with intranasally administered relevant allergens. Symptoms were recorded for 2 weeks before and 1 week after each challenge, using 3 different instruments. Blood samples were taken before, and 0, 1, 6, and 24 hours after challenges. Levels of complement split products, cell associated cytokines, and eosinophilic cationic protein were measured. Mean preexercise and postexercise symptom scores were evaluated for each group. RESULTS: Exercise challenge induced significant increases of the complement split product C4a, but not C3a or C5a, at 6 hours after exercise only in the CFS group (P <.01), regardless of allergy status. Mean symptom scores were significantly increased after exercise through the use of a daily diary (P <.03) and a weekly diary (P <.01) for the CFS group only. Mean scores for the Multidimensional Fatigue Inventory categories "reduced activity" and "mental fatigue" were significantly increased in the CFS group only (P <.04 and P <.02, respectively). CONCLUSIONS: Exercise challenge may be a valuable tool in the development of diagnostic criteria and tests for CFS. Establishment of a role for complement activation products as markers or participants in production of illness require further study. PMID- 12897749 TI - Blockade of LTC4 synthesis caused by additive inhibition of gIV-PLA2 phosphorylation: Effect of salmeterol and PDE4 inhibition in human eosinophils. AB - BACKGROUND: Prior investigations have demonstrated that beta(2)-adrenoceptor stimulation is ineffective in inhibiting synthesis of eicosanoids in human eosinophils. This effect has been postulated to relate to density or structural differences in the beta(2)-adrenoceptor or its coupled G-protein. However, recent reports indicate that cAMP-specific PDE4 activity in eosinophils is 10-fold that of other inflammatory cells. We postulated that selective blockade of PDE4 in eosinophils would unmask the inhibitory effect of beta(2)-adrenoceptor stimulation and that this inhibition would result from decreased phosphor-ylation of cytosolic group IV-PLA(2) (gIV-PLA(2)). OBJECTIVE: To determine (a) whether PDE4 inhibition alone with rolipram blocked secretions of arachidonic acid (AA) and leukotriene C(4) (LTC(4)) caused by activation of eosinophils with formyl-met leu-phe plus cytochalasin B (FMLP/B), (b) to determine if PDE4 inhibition plus beta(2)-adrenoceptor agonist act additively to augment endogenous cAMP concentration, and (c) to determine the mechanism by which additive inhibition of AA and LTC(4) synthesis is regulated by cAMP. METHODS: Human eosinophils were pretreated with buffer, salmeterol or rolipram (singly or combination) before FMLP/B activation. Release of AA and LTC(4), intracellular cAMP concentration, and phosphorylation and activation of gIV-PLA(2) were determined. RESULTS: Rolipram unmasked the inhibitory effect of beta(2)-adrenoceptor stimulation with salmeterol and significantly attenuated the stimulated release of AA and subsequent LTC(4). Inhibition corresponded to increased cAMP production caused by rolipram alone or rolipram plus salmeterol and blocked proportionately the phosphorylation and activation of gIV-PLA(2) in FMLP/B-activated eosinophils. CONCLUSIONS: Inhibition of PDE4 by rolipram unmasks beta(2)-adrenergic blockade of LTC(4) synthesis caused by FMLP/B. PMID- 12897750 TI - Surface expression of Fc epsilon RI on Langerhans' cells of clinically uninvolved skin is associated with disease activity in atopic dermatitis, allergic asthma, and rhinitis. AB - BACKGROUND: Fc epsilon RI expressed on the surface of human epidermal Langerhans' cells facilitates uptake of IgE-associated allergens and plays a pivotal role in the pathogenesis of atopic dermatitis. Seminal results from studies investigating Langerhans' cell Fc epsilon RI in skin biopsy sections or epidermal cell suspensions demonstrate the highest receptor expression in lesional skin of patients with active atopic dermatitis. OBJECTIVE: We sought to investigate and localize Fc epsilon RI expression on Langerhans' cells within a minimally disturbed tissue environment in clinically uninvolved skin and to compare receptor expression between healthy donors and patients with atopic dermatitis or other allergic diseases. METHODS: Intact epidermal sheets from skin suction blisters, immunofluorescently stained with Langerhans' cell markers and anti-Fc epsilon RI alpha (mAbs 15E5 and 22E7) or anti-IgE, were examined by means of confocal microscopy. Samples incubated with anti-Fc epsilon RI alpha before or after cell fixation-permeabilization were compared to discriminate between cytoplasmic and membrane localization. RESULTS: Cytoplasmic Fc epsilon RI alpha chain was found in Langerhans' cells from all donors, irrespective of atopic status. Surface Fc epsilon RI-bound IgE was detected in the skin of individuals with active atopic dermatitis and in the skin of those with active asthma or rhinitis. No surface Fc epsilon RI was expressed in the skin of patients with a clinical history of atopic dermatitis, asthma, or rhinitis whose disease was in remission or in the skin of nonatopic individuals. CONCLUSION: In clinically uninvolved skin, Langerhans' cell-surface Fc epsilon RI expression is not only linked to atopic dermatitis but is also generally associated with allergic disease. This supports the concept of a systemic regulatory mechanism associated with active allergic disease, which is further aggravated by local inflammation in atopic skin lesions. PMID- 12897751 TI - Is delivery by cesarean section a risk factor for food allergy? AB - BACKGROUND: Cesarean delivery might delay the colonization of the newborn intestine. A delayed or aberrant colonization process has been offered as an explanation for the increase in allergic diseases. OBJECTIVE: The aim of this study was to examine whether cesarean delivery and the use of antibiotics were associated with subsequent food allergy. METHODS: In a population-based birth cohort of 2803 children, information regarding mode of delivery, maternal or infant use of antibiotics, and information on potential confounders was obtained prospectively from parental reports and the Norwegian Birth Registry. Parentally perceived reactions to egg, fish, or nuts, as well as objectively confirmed reactions to egg at the age of 2 1/2 years, were chosen as outcomes. RESULTS: Among children whose mothers were allergic, cesarean section was associated with a 7-fold increased risk of parentally perceived reactions to egg, fish, or nuts (odds ratio, 7.0; CI, 1.8-28; P =.005) and a 4-fold increased risk of confirmed egg allergy (odds ratio, 4.1; CI, 0.9-19; P =.08) in a logistic regression analysis, adjusting for pregnancy complications, birth weight, gestational length, and socioeconomic factors. Among children whose mothers were not allergic, the association was much weaker and not significant. Maternal or infant use of antibiotics was not associated with an increased risk of food allergy. CONCLUSION: The results indicate that in predisposed children cesarean section might increase the risk of development of food allergy, which supports the theory that factors interfering with the colonization process might play a role in the development of food allergy. PMID- 12897752 TI - Detection of clinical markers of sensitization to profilin in patients allergic to plant-derived foods. AB - BACKGROUND: A proper classification of patients allergic to plant-derived foods is of pivotal importance because the clinical features of allergic reactions to fruits and vegetables depend on the nature and characteristics of proteins responsible for sensitization. However, in normal clinical settings this is presently impossible. OBJECTIVE: We sought to detect clinical markers of sensitization to profilin. METHODS: Seventy-one patients allergic to fruits and vegetables but not sensitized to lipid transfer protein or natural rubber latex were studied. Food allergy was ascertained on the basis of clinical history and positive skin prick test responses with fresh foods, commercial extracts, or both. Allergies to foods that had caused less than 2 adverse reactions were confirmed by means of open oral challenge. IgE reactivity to rBet v 1/rBet v 2 and to natural Phleum species profilin were detected. Moreover, IgE to the 30- to 40-kd and 60- to 90-kd birch pollen-enriched fractions, which also can be involved in cross-reactivity phenomena, were measured in sera from 52 patients by means of ELISA. RESULTS: On the basis of in vitro tests, 24, 18, and 25 patients turned out to be sensitized to Bet v 1, Bet v 2, or both, respectively. Four patients had negative test results for both allergens. Hypersensitivity to Bet v 2 was strongly associated with clinical allergy to citrus fruits (39% in patients monosensitized to Bet v 2 vs 4% in patients monosensitized to Bet v 1, P <.025), melon or watermelon (67% vs 0%, P <.001), banana (66% vs 8%, P <.001), and tomato (33% vs 0%, P <.05), whereas Bet v 1 sensitivity was associated with clinical allergy to apple (100% vs 39%, P <.001) and hazelnut (56% vs 0%, P <.001). The sensitivity of a history of allergy to gourd fruits, citrus fruits, tomato, banana, or a combination thereof as a means to detect profilin-hypersensitive patients was 85% (41/48). The specificity of an allergy to any of these fruits exceeded 85%, with positive predictive values ranging between 68% and 91%. CONCLUSION: In clinical settings in which laboratory investigations are not easily accessible, allergy to melon, watermelon, citrus fruits, tomato, and banana can be used as a marker of profilin hypersensitivity once a sensitization to natural rubber latex and lipid transfer protein is ruled out. PMID- 12897753 TI - Mutational analysis of major, sequential IgE-binding epitopes in alpha s1-casein, a major cow's milk allergen. AB - BACKGROUND: Allergy to cow's milk is common in early childhood, and no therapy other than avoidance exists. In murine models of peanut allergy, immunotherapy with mutated, engineered, proteins appears promising. OBJECTIVE: We sought to identify the critical amino acids (AAs) for immunoglobulin E (IgE) binding within the major B-cell epitopes of alpha(s1)-casein, a major cow's milk allergen. This will provide the necessary information to alter the cDNA to encode a protein capable of activating milk-specific T cells, but with reduced IgE-binding capacity. METHODS: For mutational analysis of the IgE-binding epitopes, peptides of 10-14 AAs in length were synthesized on a derivatized cellulose membrane with single or multiple AA substitutions. Membranes were immunolabeled with pooled sera from 15 cow's-milk-allergic patients and with 8 individual sera. RESULTS: With the pooled sera, substitution of a single AA led to complete abrogation of IgE binding to 2 of 8 peptides and diminished binding in the remainder. Substitution of multiple AAs led to an abrogation of binding in the remaining peptides. In 4 of the 8 peptides, the critical AA identified with pooled sera did not result in significant reduction of IgE binding with 1 or more individual patients. For these patients, other critical AAs were identified, indicating a more heterogeneous pattern in IgE recognition. CONCLUSION: This study indicates that single or multiple AA substitutions within IgE-binding epitopes result in reduced binding of milk-specific IgE antibodies by patients' sera. However, for future immunotherapeutic interventions with mutated peptides, critical AAs should be evaluated with individual patient sera to determine B-cell-epitope heterogeneity. PMID- 12897754 TI - The -159 C-->T polymorphism of CD14 is associated with nonatopic asthma and food allergy. AB - BACKGROUND: CD14, the receptor for LPS, plays an important role in innate immunity. A polymorphism in the promotor for CD14, -159 C-->T, has been implicated in atopy. OBJECTIVE: We explored the relationship of this polymorphism with both atopic and nonatopic asthma, as well as with food allergy. METHODS: Patients with asthma and food allergy were recruited along with nonatopic, nonasthmatic control subjects. The -159 C-->T polymorphism was genotyped by using the PCR-based RFLP assay. RESULTS: The -159 T allele was more common among patients with nonatopic asthma and food allergy than among control subjects (chi(2) = 6.03, P =.01 and chi(2) = 4.94; P =.03, respectively). Patients with food allergy had a 4-fold increased odds of having the TT genotype versus carriers of the C allele compared with control subjects (odds ratio [OR] = 3.9, 95% CI = 1.5-10.3), whereas patients with nonatopic asthma had a 3-fold increased odds of having the TT genotype (OR = 3.1 [95% CI = 1.1-9.1]). Controlling for sex differences between groups did not alter this relationship, which remained significant for patients with food allergy (OR = 3.7 [95% CI = 1.4-10.1]) or nonatopic asthma (OR = 2.7 [95% CI = 0.9-8.0]). We performed a stratified analysis, limited to white patients, to reduce population stratification. The relationship with the TT genotype was stronger in white patients with nonatopic asthma (OR = 4.4 [95% CI = 1.3-14.8]) and patients with food allergy (OR = 5.1 [95% CI = 1.6-16.2]), even adjusting for sex differences (OR = 3.9 [95% CI = 1.1 13.5] and OR = 4.6 [95% CI = 1.4-14.8], respectively). CONCLUSIONS: The TT genotype of -159 C-->T CD14 is associated with nonatopic asthma and food allergy, particularly in white subjects. Thus CD14 is a candidate gene specifically for nonatopic asthma and not for asthma in general. This indicates that atopic and nonatopic asthma might be distinct conditions in their genetic predisposition, despite the fact that they are very similar once they have been established. PMID- 12897755 TI - How and by whom care is delivered influences anti-inflammatory use in asthma: Results of a national population survey. AB - BACKGROUND: Studies examining the influence of provider behavior and patterns of care delivery on the use of anti-inflammatory asthma therapy have been limited to selected populations or have been unable to assess the appropriateness of therapy for individuals. We have previously reported the influence of sociodemographic variables and asthma severity on reported use of asthma medications in the United States. OBJECTIVE: We sought to examine the influence of patterns of care delivery and clinician behavioral factors on the use of anti-inflammatory medication by patients with asthma. METHODS: We performed a cross-sectional national random digit dial household telephone survey in 1998 of adult patients and parents of children with current asthma. Respondents were classified as having current asthma if they had a physician's diagnosis of asthma and were either taking medication for asthma or had asthma symptoms during the past year. RESULTS: One or more persons met the study criteria for current asthma in 3273 (7.8%) households in which a screening questionnaire was completed. Of the 2509 persons (721 children <16 years of age) with current asthma interviewed, 507 (20.1%) reported current use of anti-inflammatory medication. In a multiple logistic regression model controlling for asthma symptoms, reported anti inflammatory use was significantly associated with patients reporting their physician having an excellent ability to explain asthma management (odds ratio [OR], 1.47; 95% CI, 1.09-1.98), scheduling regular visits to a physician for asthma (OR, 1.30; 95% CI, 1.02-1.64), having a written asthma action plan (OR, 1.63; 95% CI, 1.29-2.06), and being of white, non-Hispanic ethnicity (OR, 1.53; 95% CI, 1.19-1.98), along with markers of greater asthma morbidity, missing 6 or more days from work or school in the past year (OR, 1.29; 95% CI, 1.01-1.65), and hospitalization for asthma in the past year (OR, 1.74; 95% CI, 1.19-2.53). Anti inflammatory use was less likely to be reported with younger age (OR, 0.82; 95% CI, 0.73-0.94), lower long-term asthma symptom burden (OR, 0.82; 95% CI, 0.71 0.94), use of 4 or fewer reliever inhaler canisters in the past year (OR, 0.50; 95% CI, 0.43-0.58), and smoking (OR, 0.50; 95% CI, 0.37-0.68). CONCLUSION: How asthma care is delivered influences the use of anti-inflammatory medication. Strategies to increase regular evaluation by a physician interested in asthma, particularly for minority patients, and to increase a physician's ability to communicate asthma management to patients might improve use of anti-inflammatory therapy among patients with asthma. PMID- 12897756 TI - Fatal posture in anaphylactic shock. PMID- 12897757 TI - Allergy to ingested mycoprotein in a patient with mold spore inhalant allergy. PMID- 12897758 TI - Association of thromboxane A2 receptor (TBXA2R) with atopy and asthma. PMID- 12897759 TI - Evidence for human mast cell nonreleaser phenotype. PMID- 12897760 TI - Lack of association between the 7888 C/T polymorphism in the AID gene and atopy in a Spanish population. PMID- 12897762 TI - Is the "hygiene hypothesis" being challenged? PMID- 12897763 TI - High-efficiency particle arrest vacuums and cat allergen exposure. PMID- 12897768 TI - Seeded conversion of recombinant prion protein to a disulfide-bonded oligomer by a reduction-oxidation process. AB - The infectious form of prion protein, PrP(Sc), self-propagates by its conversion of the normal, cellular prion protein molecule PrP(C) to another PrP(Sc) molecule. It has not yet been demonstrated that recombinant prion protein can convert prion protein molecules from PrP(C) to PrP(Sc). Here we show that recombinant hamster prion protein is converted to a second form, PrP(RDX), by a redox process in vitro and that this PrP(RDX) form seeds the conversion of other PrP(C) molecules to the PrP(RDX) form. The converted form shows properties of oligomerization and seeded conversion that are characteristic of PrP(Sc). We also find that the oligomerization can be reversed in vitro. X-ray fiber diffraction suggests an amyloid-like structure for the oligomerized prion protein. A domain swapping model involving intermolecular disulfide bonds can account for the stability and coexistence of two molecular forms of prion protein and the capacity of the second form for self-propagation. PMID- 12897767 TI - Structural basis for p38alpha MAP kinase quinazolinone and pyridol-pyrimidine inhibitor specificity. AB - The quinazolinone and pyridol-pyrimidine classes of p38 MAP kinase inhibitors have a previously unseen degree of specificity for p38 over other MAP kinases. Comparison of the crystal structures of p38 bound to four different compounds shows that binding of the more specific molecules is characterized by a peptide flip between Met109 and Gly110. Gly110 is a residue specific to the alpha, beta and gamma isoforms of p38. The delta isoform and the other MAP kinases have bulkier residues in this position. These residues would likely make the peptide flip energetically unfavorable, thus explaining the selectivity of binding. To test this hypothesis, we constructed G110A and G110D mutants of p38 and measured the potency of several compounds against them. The results confirm that the selectivity of quinazolinones and pyridol-pyrimidines results from the presence of a glycine in position 110. This unique mode of binding may be exploited in the design of new p38 inhibitors. PMID- 12897769 TI - Cooperative organization in a macromolecular complex. AB - The mechanism of assembly of multiprotein complexes and the subsequent organization of activity are not well understood. Here we report the application of biophysical tools to investigate the relationship between structure and function in protein assemblies. We used as a model system the SCF(Skp2) complex that targets p27(Kip1) for ubiquitination and subsequent degradation; this process requires an adapter protein, Cks1. By dissecting the interactions between the different subunits we show that the properties of Cks1 are highly context dependent, and its activity is acquired only when the complex is fully assembled. The results provide insights into the central role of small adapters in macromolecular assembly and explain their high sequence conservation. Simultaneous and synergistic binding of multiple subunits in a complex provides the specificity and control required before the key cell-cycle regulator p27 is committed to degradation. PMID- 12897770 TI - Control of tetrapyrrole biosynthesis by alternate quaternary forms of porphobilinogen synthase. AB - Porphobilinogen synthase (PBGS) catalyzes the first common step in the biosynthesis of tetrapyrroles (such as heme and chlorophyll). Although the predominant oligomeric form of this enzyme, as inferred from many crystal structures, is that of a homo-octamer, a rare human PBGS allele, F12L, reveals the presence of a hexameric form. Rearrangement of an N-terminal arm is responsible for this oligomeric switch, which results in profound changes in kinetic behavior. The structural transition between octamer and hexamer must proceed through an unparalleled equilibrium containing two different dimer structures. The allosteric magnesium, present in most PBGS, has a binding site in the octamer but not in the hexamer. The unprecedented structural rearrangement reported here relates to the allosteric regulation of PBGS and suggests that alternative PBGS oligomers may function in a magnesium-dependent regulation of tetrapyrrole biosynthesis in plants and some bacteria. PMID- 12897771 TI - Haplotype blocks and linkage disequilibrium in the human genome. AB - There is great interest in the patterns and extent of linkage disequilibrium (LD) in humans and other species. Characterizing LD is of central importance for gene mapping studies and can provide insights into the biology of recombination and human demographic history. Here, we review recent developments in this field, including the recently proposed 'haplotype-block' model of LD. We describe some of the recent data in detail and compare the observed patterns to those seen in simulations. PMID- 12897772 TI - The human Y chromosome: an evolutionary marker comes of age. AB - Until recently, the Y chromosome seemed to fulfil the role of juvenile delinquent among human chromosomes--rich in junk, poor in useful attributes, reluctant to socialize with its neighbours and with an inescapable tendency to degenerate. The availability of the near-complete chromosome sequence, plus many new polymorphisms, a highly resolved phylogeny and insights into its mutation processes, now provide new avenues for investigating human evolution. Y chromosome research is growing up. PMID- 12897773 TI - Technicolour transgenics: imaging tools for functional genomics in the mouse. AB - Over the past decade, a battery of powerful tools that encompass forward and reverse genetic approaches have been developed to dissect the molecular and cellular processes that regulate development and disease. The advent of genetically-encoded fluorescent proteins that are expressed in wild type and mutant mice, together with advances in imaging technology, make it possible to study these biological processes in many dimensions. Importantly, these technologies allow direct visual access to complex events as they happen in their native environment, which provides greater insights into mammalian biology than ever before. PMID- 12897774 TI - The power of the 3' UTR: translational control and development. AB - Many crucial decisions, such as the location and timing of cell division, cell fate determination, and embryonic axes establishment, are made in the early embryo, a time in development when there is often little or no transcription. For this reason, the control of variation in gene expression in the early embryo often relies on post-transcriptional control of maternal genes. Although the early embryo is rife with translational control, controlling mRNA activity is also important in other developmental processes, such as stem-cell proliferation, sex determination, neurogenesis and erythropoiesis. PMID- 12897775 TI - Genetic regulation of osteoclast development and function. AB - Osteoclasts are the principal, if not exclusive, bone-resorbing cells, and their activity has a profound impact on skeletal health. So, disorders of skeletal insufficiency, such as osteoporosis, typically represent enhanced osteoclastic bone resorption relative to bone formation. Prevention of pathological bone loss therefore depends on an appreciation of the mechanisms by which osteoclasts differentiate from their precursors and degrade the skeleton. The past five years have witnessed important insights into osteoclast formation and function. Many of these discoveries have been made through genetic experiments that involved the rare hereditary disorder osteopetrosis. PMID- 12897776 TI - Evolutionary and ecological functional genomics. AB - A unique combination of disciplines is emerging--evolutionary and ecological functional genomics--which focuses on the genes that affect ecological success and evolutionary fitness in natural environments and populations. Already this approach has provided new insights that were not available from its disciplinary components in isolation. However, future advances will necessitate the re engineering of scientific attitudes, training and institutions, to achieve extensive multidisciplinarity. PMID- 12897777 TI - An everlasting pioneer: the story of Antirrhinum research. AB - Despite the tremendous success of Arabidopsis thaliana, no single model can represent the vast range of form that is seen in the approximately 250,000 existing species of flowering plants (angiosperms). Here, we consider the history and future of an alternative angiosperm model--the snapdragon Antirrhinum majus. We ask what made Antirrhinum attractive to the earliest students of variation and inheritance, and how its use led to landmark advances in plant genetics and to our present understanding of plant development. Finally, we show how the wide diversity of Antirrhinum species, combined with classical and molecular genetics- the two traditional strengths of Antirrhinum--provide an opportunity for developmental, evolutionary and ecological approaches. These factors make A. majus an ideal comparative angiosperm. PMID- 12897778 TI - Interaction between leukemic-cell VLA-4 and stromal fibronectin is a decisive factor for minimal residual disease of acute myelogenous leukemia. AB - Bone-marrow minimal residual disease (MRD) causes relapse after chemotherapy in patients with acute myelogenous leukemia (AML). We postulate that the drug resistance is induced by the attachment of very late antigen (VLA)-4 on leukemic cells to fibronectin on bone-marrow stromal cells. We found that VLA-4-positive cells acquired resistance to anoikis (loss of anchorage) or drug-induced apoptosis through the phosphatidylinositol-3-kinase (PI-3K)/AKT/Bcl-2 signaling pathway, which is activated by the interaction of VLA-4 and fibronectin. This resistance was negated by VLA-4-specific antibodies. In a mouse model of MRD, we achieved a 100% survival rate by combining VLA-4-specific antibodies and cytosine arabinoside (AraC), whereas AraC alone prolonged survival only slightly. In addition, overall survival at 5 years was 100% for 10 VLA-4-negative patients and 44.4% for 15 VLA-4-positive patients. Thus, the interaction between VLA-4 on leukemic cells and fibronectin on stromal cells may be crucial in bone marrow MRD and AML prognosis. PMID- 12897779 TI - Cyclophilin A modulates the sensitivity of HIV-1 to host restriction factors. AB - Many mammalian species express restriction factors that confer host resistance to retroviral infection. Here we show that HIV-1 sensitivity to restriction factors is modulated by cyclophilin A (CypA), a host cell protein that binds the HIV-1 capsid protein (CA). In certain nonhuman primate cells, the CA-CypA interaction is essential for restriction: HIV-1 infectivity is increased >100-fold by cyclosporin A (CsA), a competitive inhibitor of the interaction, or by an HIV-1 CA mutation that disrupts CypA binding. Conversely, disruption of CA-CypA interaction in human cells reveals that CypA protects HIV-1 from the Ref-1 restriction factor. These findings suggest that HIV-1 has co-opted a host cell protein to counteract restriction factors expressed by human cells and that this adaptation can confer sensitivity to restriction in unnatural hosts. Manipulation of HIV-1 CA recognition by restriction factors promises to advance animal models and new therapeutic strategies for HIV-1 and AIDS. PMID- 12897780 TI - The nonimmunoglobulin portion of lambda5 mediates cell-autonomous pre-B cell receptor signaling. AB - The pre-B cell receptor (preBCR), composed of mu immunoglobulin (Ig) and surrogate light chains, signals large 'preB-II' cells to proliferate in the apparent absence of ligands or cooperating cells. We deleted the N-terminal, nonimmunoglobulin (nonlg) portion of lambda5, or mutated seven arginine residues in it to serine residues. PreBCRs with such mutant lambda5 proteins showed increased cell surface representation and a diminished rate of aggregation and internalization. Tyrosine phosphorylation of preBCR complexes containing mutant lambda5 proteins was abolished. These results indicate that the nonIg portion of lambda5, and the seven arginine residues in it, are needed for signal transduction, and that signaling could be cell autonomous. We propose two models to explain the apparently constitutive, ligand-independent signal-transducing capacity of the preBCR. PMID- 12897781 TI - Crystal structure of HLA-A2 bound to LIR-1, a host and viral major histocompatibility complex receptor. AB - Leukocyte immunoglobulin-like receptor 1 (LIR-1), an inhibitory receptor expressed on monocytes, dendritic cells and lymphocytes, regulates cellular function by binding a broad range of classical and nonclassical major histocompatibility complex (MHC) class I molecules, and the human cytomegalovirus MHC class I homolog UL18. Here we describe the 3.4-A crystal structure of a complex between the LIR-1 D1D2 domains and the MHC class I molecule HLA-A2. LIR-1 contacts the mostly conserved beta(2)-microglobulin and alpha3 domains of HLA-A2. The LIR-1 binding site comprises residues at the interdomain hinge, and a patch at the D1 tip. The structure shows how LIR-1 recognizes UL18 and diverse MHC class I molecules, and indicates that a similar mode of MHC class I recognition is used by other LIR family members. PMID- 12897782 TI - Trans-acting regulatory variation in Saccharomyces cerevisiae and the role of transcription factors. AB - Natural genetic variation can cause significant differences in gene expression, but little is known about the polymorphisms that affect gene regulation. We analyzed regulatory variation in a cross between laboratory and wild strains of Saccharomyces cerevisiae. Clustering and linkage analysis defined groups of coregulated genes and the loci involved in their regulation. Most expression differences mapped to trans-acting loci. Positional cloning and functional assays showed that polymorphisms in GPA1 and AMN1 affect expression of genes involved in pheromone response and daughter cell separation, respectively. We also asked whether particular classes of genes were more likely to contain trans-regulatory polymorphisms. Notably, transcription factors showed no enrichment, and trans regulatory variation seems to be broadly dispersed across classes of genes with different molecular functions. PMID- 12897783 TI - LINE-mediated retrotransposition of marked Alu sequences. AB - Alu elements are the most successful transposons in humans. They are 300-bp non coding sequences transcribed by RNA polymerase III (Pol III) and are expected to retrotranspose with the aid of reverse transcriptases of cellular origin. We previously showed that human LINEs can generate cDNA copies of any mRNA transcript by means of a retroposition process involving reverse transcription and integration by the LINE-encoded endonuclease and reverse transcriptase. Here we show mobility of marked Alu sequences in human HeLa cells with the canonical features of a retrotransposition process, including splicing out of an autocatalytic intron introduced into the marked sequence, target site duplications of varying lengths and integrations into consensus A-rich sequences. We further show that the poly-A stretch at the Alu 3' end is essential for mobility, that LINEs are required for transposition and that the rate of retroposition is 100-1,000 times higher for Alu transcripts than for control mRNAs, thus accounting for the high mutational activity of these elements observed in humans. PMID- 12897784 TI - Mutations associated with neutropenia in dogs and humans disrupt intracellular transport of neutrophil elastase. AB - Cyclic hematopoiesis is a stem cell disease in which the number of neutrophils and other blood cells oscillates in weekly phases. Autosomal dominant mutations of ELA2, encoding the protease neutrophil elastase, found in lysosome-like granules, cause cyclic hematopoiesis and most cases of the pre-leukemic disorder severe congenital neutropenia (SCN; ref. 3) in humans. Over 20 different mutations of neutrophil elastase have been identified, but their consequences are elusive, because they confer no consistent effects on enzymatic activity. The similar autosomal recessive disease of dogs, canine cyclic hematopoiesis, is not caused by mutations in ELA2 (data not shown). Here we show that homozygous mutation of the gene encoding the dog adaptor protein complex 3 (AP3) beta subunit, directing trans-Golgi export of transmembrane cargo proteins to lysosomes, causes canine cyclic hematopoiesis. C-terminal processing of neutrophil elastase exposes an AP3 interaction signal responsible for redirecting neutrophil elastase trafficking from membranes to granules. Disruption of either neutrophil elastase or AP3 perturbs the intracellular trafficking of neutrophil elastase. Most mutations in ELA2 that cause human cyclic hematopoiesis prevent membrane localization of neutrophil elastase, whereas most mutations in ELA2 that cause SCN lead to exclusive membrane localization. PMID- 12897785 TI - Afferent modulation of dopamine neuron firing differentially regulates tonic and phasic dopamine transmission. AB - The mesolimbic dopamine system is centrally involved in reward and goal-directed behavior, and it has been implicated in multiple psychiatric disorders. Understanding the mechanism by which dopamine participates in these activities requires comprehension of the dynamics of dopamine release. Here we report dissociable regulation of dopamine neuron discharge by two separate afferent systems in rats; inhibition of pallidal afferents selectively increased the population activity of dopamine neurons, whereas activation of pedunculopontine inputs increased burst firing. Only the increase in population activity increased ventral striatal dopamine efflux. After blockade of dopamine reuptake, however, enhanced bursting increased dopamine efflux three times more than did enhanced population activity. These results provide insight into multiple regulatory systems that modulate dopamine system function: burst firing induces massive synaptic dopamine release, which is rapidly removed by reuptake before escaping the synaptic cleft, whereas increased population activity modulates tonic extrasynaptic dopamine levels that are less influenced by reuptake. PMID- 12897786 TI - Lmx1b is essential for the development of serotonergic neurons. AB - The specification and differentiation of serotonergic (5-HT) neurons require both extrinsic signaling molecules and intrinsic transcription factors to work in concert or in cascade. Here we identify the genetic cascades that control the specification and differentiation of 5-HT neurons in mice. A major determinant in the cascades is an LIM homeodomain-containing gene, Lmx1b, which is required for the development of all 5-HT neurons in the central nervous system. Our results suggest that, during development of 5-HT neurons, Lmx1b is a critical intermediate factor that couples Nkx2-2-mediated early specification with Pet1 mediated terminal differentiation. Moreover, our data indicate that genetic cascades controlling the caudal and rostral 5-HT neurons are distinct, despite their shared components. PMID- 12897787 TI - Alternative splicing of lola generates 19 transcription factors controlling axon guidance in Drosophila. AB - The Drosophila melanogaster transcription factor Lola (longitudinals lacking) is a pivotal regulator of neural wiring that sets the precise expression levels of proteins that execute specific axon guidance decisions. Lola has a zinc finger DNA binding domain and a BTB (for Broad-complex, Tramtrack and Bric a brac) dimerization motif. We now show that alternative splicing of the lola gene creates a family of 19 transcription factors. All lola isoforms share a common dimerization domain, but 17 have their own unique DNA-binding domains. Seven of these 17 isoforms are present in the distantly-related Dipteran Anopheles gambiae, suggesting that the properties of specific isoforms are likely to be critical to lola function. Analysis of the expression patterns of individual splice variants and of the phenotypes of mutants lacking single isoforms supports this idea and establishes that the alternative forms of lola are responsible for different functions of this gene. Thus, in this system, the alternative splicing of a key transcription factor helps to explain how a small genome encodes all the information that is necessary to specify the enormous diversity of axonal trajectories. PMID- 12897788 TI - Adenylyl cyclase I regulates AMPA receptor trafficking during mouse cortical 'barrel' map development. AB - Cortical map formation requires the accurate targeting, synaptogenesis, elaboration and refinement of thalamocortical afferents. Here we demonstrate the role of Ca2+/calmodulin-activated type-I adenylyl cyclase (AC1) in regulating the strength of thalamocortical synapses through modulation of AMPA receptor (AMPAR) trafficking using barrelless mice, a mutant without AC1 activity or cortical 'barrel' maps. Barrelless synapses are stuck in an immature state that contains few functional AMPARs that are rarely silent (NMDAR-only). Long-term potentiation (LTP) and long-term depression (LTD) at thalamocortical synapses require postsynaptic protein kinase A (PKA) activity and are difficult to induce in barrelless mice, probably due to an inability to properly regulate synaptic AMPAR trafficking. Consistent with this, both the extent of PKA phosphorylation on AMPAR subunit GluR1 and the expression of surface GluR1 are reduced in barrelless neurons. These results suggest that activity-dependent mechanisms operate through an AC1/PKA signaling pathway to target some synapses for consolidation and others for elimination during barrel map formation. PMID- 12897789 TI - Behavioral detection of tactile stimuli during 7-12 Hz cortical oscillations in awake rats. AB - Prominent 7-12 Hz oscillations in the primary somatosensory cortex (S1) of awake but immobile rats might represent a seizure-like state in which neuronal burst firing renders animals unresponsive to incoming tactile stimuli; others have proposed that these oscillations are analogous to human mu rhythm. To test whether rats can respond to tactile stimuli during 7-12 Hz oscillatory activity, we trained head-immobilized awake animals to indicate whether they could detect the occurrence of transient whisker deflections while we recorded local field potentials (LFPs) from microelectrode arrays implanted bilaterally in the S1 whisker representation area. They responded rapidly and reliably, suggesting that this brain rhythm represents normal physiological activity that does not preclude perception. PMID- 12897790 TI - Metabolic redesign of vitamin E biosynthesis in plants for tocotrienol production and increased antioxidant content. AB - Tocotrienols are the primary form of vitamin E in seeds of most monocot plants, including cereals such as rice and wheat. As potent antioxidants, tocotrienols contribute to the nutritive value of cereal grains in human and livestock diets. cDNAs encoding homogentisic acid geranylgeranyl transferase (HGGT), which catalyzes the committed step of tocotrienol biosynthesis, were isolated from barley, wheat and rice seeds. Transgenic expression of the barley HGGT in Arabidopsis thaliana leaves resulted in accumulation of tocotrienols, which were absent from leaves of nontransformed plants, and a 10- to 15-fold increase in total vitamin E antioxidants (tocotrienols plus tocopherols). Overexpression of the barley HGGT in corn seeds resulted in an increase in tocotrienol and tocopherol content of as much as six-fold. These results provide insight into the genetic basis for tocotrienol biosynthesis in plants and demonstrate the ability to enhance the antioxidant content of crops by introduction of an enzyme that redirects metabolic flux. PMID- 12897792 TI - Molecular medicine: trendy title or new reality? PMID- 12897791 TI - Random peptide libraries displayed on adeno-associated virus to select for targeted gene therapy vectors. AB - Characterizing the molecular diversity of the cell surface is critical for targeting gene therapy. Cell type-specific binding ligands can be used to target gene therapy vectors. However, targeting systems in which optimum eukaryotic vectors can be selected on the cells of interest are not available. Here, we introduce and validate a random adeno-associated virus (AAV) peptide library in which each virus particle displays a random peptide at the capsid surface. This library was generated in a three-step system that ensures encoding of displayed peptides by the packaged DNA. As proof-of-concept, we screened AAV-libraries on human coronary artery endothelial cells. We observed selection of particular peptide motifs. The selected peptides enhanced transduction in coronary endothelial cells but not in control nonendothelial cells. This vector targeting strategy has advantages over other combinatorial approaches such as phage display because selection occurs within the context of the capsid and may have a broad range of applications in biotechnology and medicine. PMID- 12897793 TI - Cover the eyes of Lady Justice. PMID- 12897794 TI - A global player for public health. An interview with Tikki Pang, Director of Research Policy and Cooperation at the World Health Organization [interviewed by Holger Breithaupt and Caroline Hadley]. PMID- 12897795 TI - Revolution or evolution? PMID- 12897796 TI - Breathing new life into the biology classroom. PMID- 12897797 TI - Attack on amyloid. PMID- 12897798 TI - Coupling the cell cycle to cell growth. AB - In order to multiply, both prokaryotic and eukaryotic cells go through a series of events that are collectively called the cell cycle. However, DNA replication, mitosis and cell division may also be viewed as having their own, in principle independent, cycles, which are tied together by mechanisms extrinsic to the cell cycle--the checkpoints--that maintain the order of events. We propose that our understanding of cell-cycle regulation is enhanced by viewing each event individually, as an independently regulated process. The nature of the parameters that regulate cell-cycle events is discussed and, in particular, we argue that cell mass is not such a parameter. PMID- 12897799 TI - Gli proteins and the control of spinal-cord patterning. AB - The secreted protein sonic hedgehog (Shh) is crucial for the specification of neuronal subtype identity in the vertebrate neural tube. Zinc-finger proteins of the Gli family are known to be transcriptional mediators of Shh signalling, and to coordinately pattern the dorsal-ventral axis of the spinal cord. Recent studies indicate that additional signals may provide positional information in parallel to Shh to specify neuronal fate in this tissue. We review the role of Gli proteins in spinal-cord development and propose that various upstream patterning signals may be integrated by the Gli proteins to direct a coherent programme of neurogenesis. PMID- 12897801 TI - Allosteric effects mediate CHK2 phosphorylation of the p53 transactivation domain. AB - The tumour suppressor p53 is a tetrameric protein that is phosphorylated in its BOX-I transactivation domain by checkpoint kinase 2 (CHK2) in response to DNA damage. CHK2 cannot phosphorylate small peptide fragments of p53 containing the BOX-I motif, indicating that undefined determinants in the p53 tetramer mediate CHK2 recognition. Two peptides derived from the DNA-binding domain of p53 bind to CHK2 and stimulate phosphorylation of full-length p53 at Thr 18 and Ser 20, thus identifying CHK2-docking sites. CHK2 can be fully activated in trans by the two p53 DNA-binding-domain peptides, and can phosphorylate BOX-I transactivation domain fragments of p53 at Thr 18 and Ser 20. Although CHK2 has a basal Ser 20 kinase activity that is predominantly activated towards Thr 18, CHK1 has constitutive Thr 18 kinase activity that is predominantly activated in trans towards Ser 20. Cell division cycle 25C (CDC25C) phosphorylation by CHK2 is unaffected by the p53 DNA-binding-domain peptides. The CHK2-docking site in the BOX-V motif is the smallest of the two CHK2 binding sites, and mutating certain amino acids in the BOX-V peptide prevents CHK2 activation. A database search identified a p53 BOX-I-homology motif in p21(WAF1) and although CHK2 is inactive towards this protein, the p53 DNA-binding-domain peptides induce phosphorylation of p21(WAF1) at Ser 146. This provides evidence that CHK2 can be activated allosterically towards some substrates by a novel docking interaction, and identify a potential regulatory switch that may channel CHK2 into distinct signalling pathways in vivo. PMID- 12897800 TI - Effect of CpG methylation on RAG1/RAG2 reactivity: implications of direct and indirect mechanisms for controlling V(D)J cleavage. AB - It has been suggested that DNA methylation/demethylation is involved in regulating V(D)J rearrangement. Although methylated DNA is thought to induce an inaccessible chromatin structure, it is unclear whether DNA methylation can directly control V(D)J recombination independently of chromatin structure. In this study, we tested whether DNA methylation directly affects the reactivity of the RAG1/RAG2 complex. Specific methylation within the heptamer of the recombination signal sequences (RSS) markedly reduced V(D)J cleavage without inhibiting RAG1/RAG2-DNA complex formation. By contrast, methylation at other positions around the RSS did not affect the reactivity of the RAG proteins. The presence of a methyl-CpG binding-domain protein inhibited the binding of the RAG1/RAG2 complex to all the methylated CpG sites that were tested. Our findings suggest that DNA methylation around the RSS may have a previously unexpected function in regulating V(D)J recombination by directly inhibiting V(D)J cleavage, in addition to its general function of inducing an inaccessible chromatin configuration. PMID- 12897802 TI - Integrin-alpha3 mediates binding of Chordin to the cell surface and promotes its endocytosis. AB - Dorsoventral patterning in animal development is regulated by a morphogenetic gradient of Bone morphogenetic protein signalling, which is established by a set of proteins that are conserved from Drosophila to vertebrates. These include Chordin (Chd)/Short gastrulation, Xolloid/Tolloid and Twisted gastrulation. Here, we report the identification of a cell-surface component of this morphogenetic pathway. Prompted by the observation that Chd protein bound to the surface of certain cell lines with subnanomolar affinity, we identified two cell-surface proteins that bind to Chd, one of which corresponds to Integrin-alpha3. Integrin alpha3 and Chd are co-expressed in the Xenopus embryo. Transfection of Integrin alpha3 increased the binding of Chd to the cell surface, which was competed by an excess of soluble Integrin-alpha3. After binding to the cell surface, Chd was translocated into intracellular endocytic compartments in a temperature-dependent manner. We propose that Integrin-alpha3 may regulate the concentration of Chd protein in the extracellular space by endocytosis. PMID- 12897803 TI - Inotropic effects on mammalian skeletal muscle change with contraction frequency. AB - Over the last decade, we have attempted to determine if mammalian skeletal muscle's steady-level force development as established by mechanical and stimulation parameters can be increased or decreased by physiological signals. In these experiments, nitric oxide (NO), endothelin-1 (ET-1), adenosine (Ado), and beta-adrenergic agonists (beta) modified force production in the soleus and (or) the extensor digitorum longus (EDL) of the mouse. NO and beta increased the force produced by 0.5-s tetanic contractions at 0.6 contractions/min in both muscles. While EDL did not respond to either Ado or ET-1, the developed force of the soleus was amplified by Ado but attenuated by ET-1. Increased cAMP analogue concentrations amplified developed force in both muscles, but a cGMP analogue had no effect on either muscle. Following an increase in the contraction frequency of the soleus, the increased force in response to NO disappeared, as did the decreased force to ET-1. The increase in force due to a cAMP analogue disappeared during fatigue but reappeared quickly during recovery. Thus, steady-level developed force can be modified by a number of substances that can be released from locations in the body or muscle. The response to a given compound is determined by a complex interaction of metabolic and intracellular signals on the force-generating cascade. PMID- 12897804 TI - The physiology of S-nitrosothiols: carrier molecules for nitric oxide. AB - Recent work has demonstrated that inhalation of nitric oxide (NO) can impact the peripheral vasculature, suggesting that an NO-stabilizing moiety may exist in vivo. One possibility is the formation of S-nitrosothiols, which extend the half life of NO manyfold. In this review, we provide evidence that S-nitrosothiols exist in the vasculature, particularly during NO inhalation. The potential biochemical pathways that have been proposed for the formation of these products are also summarized. Finally, we highlight the limited evidence for the role that these potent vasodilating molecules may play as physiologically and therapeutically important regulators of the vascular system. PMID- 12897805 TI - Adaptation in the respiratory control system. AB - Exposure to hypoxia, whether for short or prolonged periods or for repeated episodes, produces alterations in the ventilatory responses. This review presents evidence that these adaptations are likely to be mediated by adaptations in the respiratory chemoreflexes, particularly the peripheral chemoreflex, and proposes models of respiratory control explaining the observed changes in ventilation. After a brief introduction to the respiratory control system, a graphical model is developed that illustrates the operation of the system in the steady state, which will be used later. Next, the adaptations in ventilatory responses to hypoxia that have been observed are described, and methods of measuring the alterations in the chemoreflexes that might account for them are discussed. Finally, experimental data supporting the view that changes in the activity of the peripheral chemoreflex can account for the ventilatory adaptations to hypoxia are presented and incorporated into models of chemoreflex behaviour during exposures to hypoxia of various durations. PMID- 12897806 TI - The essential role of carotid body chemoreceptors in sleep apnea. AB - Sleep apnea is attributable, in part, to an unstable ventilatory control system and specifically to a narrowed "CO2 reserve" (i.e., the difference in P(a)CO2 between eupnea and the apneic threshold). Findings from sleeping animal preparations with denervated carotid chemoreceptors or vascularly isolated, perfused carotid chemoreceptors demonstrate the critical importance of peripheral chemoreceptors to the ventilatory responses to dynamic changes in P(a)CO2. Specifically, (i) carotid body denervation prevented the apnea and periodic breathing that normally follow transient ventilatory overshoots; (ii) the CO2 reserve for peripheral chemoreceptors was about one half that for brain chemoreceptors; and (iii) hypocapnia isolated to the carotid chemoreceptors caused hypoventilation that persisted over time despite a concomitant, progressive brain respiratory acidosis. Observations in both humans and animals are cited to demonstrate the marked plasticity of the CO2 reserve and, therefore, the propensity for apneas and periodic breathing, in response to changing background ventilatory stimuli. PMID- 12897807 TI - Mechanism of the negative inotropic effects of alpha 1-adrenoceptor agonists on mouse myocardium. AB - This study was done to identify the mechanism of the alpha1-adrenoceptor (AR) mediated negative inotropic effects of phenylephrine (PE) on adult mouse myocardium. As reported by others, we also found that the nonselective alpha1AR agonist PE produced a negative inotropic effect on ventricular strips from adult mice that was inhibited by the alpha1AAR antagonist 5-methylurapidil (5MU) but not by the alpha1BAR antagonist chloroethylclonidine (CEC) or the alpha1DAR antagonist BMY 7378. The selective alpha1AAR agonist A61603 also produced a negative inotropic effect, which was antagonized by 5MU. Phorbol 12,13-dibutyrate (activator of all PKC isoforms) mimicked the negative inotropic responses to PE and A61603. The negative inotropic effects of PE were inhibited by bisindolylmaleimide (inhibitor of all PKC isoforms) but not by Go 6976 (inhibitor of Ca2+-dependent PKC). Rottlerin, an inhibitor of Ca2+-independent PKCdelta, antagonized the negative inotropic effects of PE and A61603. PE and A61603 increased the translocation of PKCdelta, which was prevented by rottlerin. These data suggest that the alpha1AR-mediated negative inotropy on adult mouse myocardium is signaled by Ca2+-independent PKCdelta. PMID- 12897808 TI - Functional study of intracellular P-gp- and MRP1-mediated pumping of free cytosolic pirarubicin into acidic organelles in intrinsic resistant SiHa cells. AB - We sought to determine the efficiency of the intracellular functional P-gp- and MRP1-mediated pumping of THP into acidic organelles in SiHa cells and etoposide resistant SiHa/VP16 cells. The expression of both MDR1 and MRP1 genes of SiHa and SiHa/VP16 cells was clearly shown by using RT-PCR. The functional studies of both intracellular functional P-gp- and MRP1-mediated pumping were performed by using THP in a conventional spectrofluorometer, and they demonstrated that SiHa and SiHa/VP16 cells are good models to illustrate the functional role of intracellular P-gp and MRP1 in the transport of free cytosolic drug into acidic organelles. The functional P-gp and MRP1 proteins were identified both on plasma membranes and on intracellular vesicle membranes. Within the limit of experimental error, similar efficiencies in THP transport were observed in the two proteins at both locations in SiHa and SiHa/VP16 cells. The P-gp- and MRP1 mediated pump coefficient (k v a), Michealis-Menten's constant (K V m), and maximal pumping rate (V V max) values of those located on vesicular membranes were 1.87 +/- 0.30 pL x cell-1 x s-1, 1.63 +/- 0.21 microM, and 4.95 +/- 0.45 nM x s-1, respectively. Drug retention inside acidic organelles (C mon V) of SiHa cells was significantly higher than that of SiHa/VP16 cells, perhaps a consequence of slower movement of recycling endosomes and (or) lysosomes to the cell membrane of SiHa cells, leading to distended organelles and cell death. Our results suggest that intracellular P-gp and MRP1 proteins play an important role in the transport of free drug from cytosol to cytoplasmic acidic organelles. PMID- 12897809 TI - Verapamil metabolites: potential P-glycoprotein-mediated multidrug resistance reversal agents. AB - Multidrug resistance in cancer chemotherapy frequently correlates with overexpression of the P-glycoprotein drug transporter. Attempts to reverse P glycoprotein-mediated multidrug resistance with racemic verapamil or its less toxic (R)-enantiomer have been complicated by cardiotoxicity. The objective of this study was to investigate the effects of the major verapamil metabolite, norverapamil, as well as the PR-22 and D-620 metabolites, on P-glycoprotein mediated drug transport. We measured the basolateral-to-apical fluxes of the P glycoprotein substrates digoxin and vinblastine in the presence and absence of verapamil, (R)-norverapamil, (S)-norverapamil, racemic norverapamil, PR-22, or D 620 across confluent monolayers of Madin-Darby canine kidney (MDCK) cells that express P-glycoprotein on their apical membranes. Verapamil and norverapamil nonstereospecifically inhibited the renal tubular secretion of digoxin and vinblastine similarly in a dose-dependent manner. However, there was no decrease in the cellular accumulation of digoxin and vinblastine, suggesting that neither verapamil nor norverapamil prevent the substrates from entering the MDCK cells. Furthermore, the norverapamil metabolite P-22 also inhibited the secretion of these P-glycoprotein substrates. Our results suggest that the verapamil metabolites norverapamil and PR-22, which are less cardiotoxic than the parent compound, have comparable inhibitory abilities to verapamil (norverapamil greater than PR-22) and may be useful in reversing resistance to P-glycoprotein substrates. PMID- 12897810 TI - Effects of barbiturates on human platelet aggregation differ depending on their chemical structures. AB - The effects of barbiturates on human platelet function are not fully understood. Since we have already revealed the effects and mechanisms of thiopental, thiamylal, and pentobarbital in platelets, the present study attempted to elucidate (i) the effects of other barbiturates on human platelet aggregation, (ii) the underlying mechanisms, and (iii) the structure-function relationship of barbiturates in platelets. Barbiturates, including amobarbital, butalbital, secobarbital, barbital, phenobarbital, metharbital, and primidone, were examined. Human platelet aggregation induced by adenosine diphosphate (ADP), epinephrine, and (+)-9,11-epithia-11,12-methano-thromboxane A2 (STA2), a thromboxane A2 analog, was measured using an 8-channel light-transmission aggregometer. The cytosolic free calcium concentration ([Ca2+]i) was measured by fluorometer using fura-2 loaded platelets. Inositol 1,4,5-trisphosphate (IP3) formation induced by STA2 was determined by a commercially available IP3 assay kit. Amobarbital, butalbital, and secobarbital suppressed ADP-, epinephrine- and STA2-induced platelet aggregation and the STA2-induced [Ca2+]i increase, even when Ca2+ influx was blocked by Ni2+. However, they did not affect STA2-induced IP3 formation. Barbital, phenobarbital, metharbital, and primidone (up to 1 mM) had no effect on ADP- and epinephrine-induced platelet aggregation. Thus, we conclude that amobarbital, butalbital, and secobarbital inhibit platelet aggregation by suppressing [Ca2+]i increase without affecting IP3 formation. However, these antiaggregatory effects may not have clinical importance, since the barbiturate concentrations used were higher than clinically relevant ones. The other tested barbiturates had no effects on platelet aggregation. The data indicate that the effects of barbiturates on platelet aggregation differ depending on their chemical structures. PMID- 12897811 TI - Role of metallothionein in regulating the abundance of histochemically reactive zinc in rat tissues. AB - The objectives of this study were (i) to investigate the modulating effects of zinc nutrition on histochemically reactive zinc in the rat intestine and liver and (ii) to assess the relationship between histochemically reactive zinc and metallothionein-bound zinc in these tissues under varying zinc nutrition. Male Wistar rats were fed a zinc-deficient (3 mg zinc/kg), adequate-zinc (30 mg zinc/kg, ad libitum or pair-fed), or zinc-supplemented (155 mg zinc/kg) diet for 2 or 6 weeks. Plasma N-(6-methoxy-8-quinolyl)-para-toluenesulfonamide-reactive zinc reflected dietary zinc intake. Abundance of the intestine histochemically reactive zinc was correlated with dietary zinc intake after 2 weeks of dietary treatment. Dietary zinc intake had no effect on the abundance of the intestine histochemically reactive zinc after 6 weeks of dietary treatment and the hepatic histochemically reactive zinc after both 2 and 6 weeks of dietary treatment. This lack of effect of dietary zinc intake on the abundance of histochemically reactive zinc was associated with a higher level of metallothionein. The molecular-mass distribution profile revealed that N-(6-methoxy-8-quinolyl)-para toluenesulfonamide-reactive zinc and metallothionein-bound zinc represented two different, but interrelated, pools of zinc. Overall, these results suggested that the abundance of histochemically reactive zinc was homeostatically regulated, which was partially achieved through the regulation of metallothionein levels in rats. PMID- 12897812 TI - Effects of Pb2+ on the transient outward potassium current in acutely dissociated rat hippocampal neurons. AB - Modulation of the voltage-dependent transient outward potassium current (IA) by Pb2+ was studied in acutely dissociated rat hippocampal pyramidal cells from the CA1 region at postnatal ages 7-14 days using the conventional whole-cell patch clamp technique. In the presence of different concentrations of external Pb2+, the initial delay and activation time of IA were concentration-dependently lengthened. In particular, the initial delay was even longer in 1 mM Pb2+, showing no signs of saturation. Pb2+ also slowed the inactivation of IA, for decay time constants in the presence of Pb2+ were increased under the same experimental protocols. The activation curves, which were reasonably fitted by a single Boltzmann function, illustrated that Pb2+ increased the voltage threshold of IA and shifted the normalized activation current-voltage curves to more depolarizing voltage commands. Moreover, Pb2+ significantly affected the steady state inactivation of IA. The application of Pb 2+ made the curves of the steady state inactivation of IA shift to more depolarizing voltages with little change in the slopes factors. In brief, the results demonstrated that Pb2+ is a dose- and voltage-dependent, reversible blocker of IA currents of hippocampal CA1 neurons. The observations were fitted by the revised "Kuo and Chen type model", which postulates a Pb2+-selective site near the pore of the IA channel and that modulation of the IA channel by Pb2+ is the result of the competitive influences of Pb2+ on opening and inactivating different pathways. PMID- 12897813 TI - Inhibitors of nitric oxide synthesis block cold-induced thermogenesis in rats. AB - N-nitro-arginine methyl ester (L-NAME), an unspecific nitric oxide synthase inhibitor, was administered to individually caged Sprague-Dawley rats exposed to cold (18 degrees C) and thermoneutral (30 degrees C) environmental temperatures during the active phase of the animals' circadian cycle. Unrestrained rats were administered intraperitoneal injections of 100 mg x kg-1 L-NAME or 1 mL x kg-1 saline. Telemetry was used to measure abdominal temperature. On a separate occasion, metabolic rate and evaporative water loss were measured using indirect calorimetery, before and after the injection of 100 mg x kg-1 L-NAME, in rats exposed to the two environments. Injection of L-NAME had no significant effect on body temperature, metabolic rate, or evaporative water loss in rats exposed to the 30 degrees C environment. In the 18 degrees C environment, L-NAME injection caused a prolonged fall in body temperature ( F(1,12) = 17.43, P = 0.001) and a significant decrease in metabolic rate (Student's t test, P = 0.001) and evaporative water loss (one-sample t test, P = 0.04). Therefore, the effects that systemic injection of L-NAME has on body temperature are dependent on environmental temperature, with nitric oxide synthase inhibition seemingly preventing the metabolic component of cold defence. PMID- 12897814 TI - Effects and consequences of nerve injury on the electrical properties of sensory neurons. AB - Nociceptive pain alerts the body to potential or actual tissue damage. By contrast, neuropathic or "noninflammatory" pain, which results from injury to the nervous system, serves no useful purpose. It typically continues for years after the original injury has healed. Sciatic nerve lesions can invoke chronic neuropathic pain that is accompanied by persistent, spontaneous activity in primary afferent fibers. This activity, which reflects changes in the properties and functional expression of Na+, K+, and Ca2+ channels, initiates a further increase in the excitability of second-order sensory neurons in the dorsal horn. This change persists for many weeks. The source of origin of the pain thus moves from the peripheral to the central nervous system. We hypothesize that this centralization of pain involves the inappropriate release of peptidergic neuromodulators from primary afferent fibers. Peptides such as substance P, neuropeptide Y (NPY), calcitonin-gene-related peptide (CGRP), and brain-derived neurotrophic factor (BDNF) may promote enduring changes in excitability as a consequence of neurotrophic actions on ion channel expression in the dorsal horn. Findings that form the basis of this hypothesis are reviewed. Study of the neurotrophic control of ion channel expression by spinal peptides may thus provide new insights into the etiology of neuropathic pain. PMID- 12897815 TI - Possible role of nicaraven in neuroprotective effect on hippocampal slice culture. AB - Nicaraven is an agent that is especially beneficial in vasospasm or brain damage caused by subarachnoid hemorrhage. It ameliorates neurological deficits of patients and protects the central nervous system from ischemia. We investigated the neuroprotective effect of nicaraven against oxygen-glucose deprivation (OGD) induced or N-methyl-D-aspartic acid (NMDA) induced hippocampal neuronal cell death in organotypic brain slice cultures. The effect of nicaraven on hippocampal neuronal injury was evaluated by inhibition of uptake of propidium iodide (PI) into dead cells. The results demonstrated that nicaraven protected neuronal cells from both OGD- and NMDA-induced cell death. While nicaraven has a strong hydroxyl radical scavenging effect, another radical scavenger, N-acetyl-L-cysteine (NAC), inhibited cell death only caused by OGD. In contrast, the poly(ADP-ribose) synthetase (PARS) inhibitors 3-aminobenzamide (3-AB) and theophylline protected cells from both OGD- and NMDA-induced cell death. Since nicaraven has an inhibitory effect in PARS, as well as a radical scavenging effect, these results suggest that inhibition of hippocampal cell death caused by NMDA may be attributable to PARS inhibition by nicaraven. PMID- 12897816 TI - Protection from solar simulated radiation-induced DNA damage in cultured human fibroblasts by three commercially available sunscreens. AB - Exposure to solar radiation can produce both acute and chronic changes in the skin, including sunburn, edema, immunosuppression, premature skin aging, and skin cancer. At the cellular level, solar radiation can produce adverse structural and functional changes in membrane proteins and lipids and in chromosomal and mitochondrial DNA. The increasing awareness of these adverse effects has led the public to demand better photoprotection. In this study, the alkaline comet assay was used to evaluate the photoprotective effects of three commercially available sunscreens at sun protection factors (SPF) 15 and 30. Human fibroblasts were used as target cells to conveniently study the effects of solar simulated radiation on DNA damage in the presence and absence of sunscreens. When human fibroblasts were exposed to various doses of solar simulated radiation, DNA damage, as measured in sunscreen-protected cells by the comet assay, was not significantly different from that detected in unexposed cells. At 1.0 and 1.5 minimal erythemal doses (MED), all sunscreens, at both SPF 15 and 30, provided nearly 100% photoprotection to the fibroblasts. Further studies are required to elucidate the role of UVA in the production and repair of DNA damage in cells exposed to sunlight. PMID- 12897817 TI - Aerobic conditioning effects on substrate responses during graded cycling in pregnancy. AB - This study was conducted to test the hypothesis that aerobic conditioning prevents exercise-induced hypoglycemia and preserves the capacity to utilize carbohydrates and to produce lactate during heavy exercise in late gestation. The effects of closely monitored cycle ergometer conditioning (heart rate = 143 +/- 2 beats/min, 25 min/day, 3 days/week) during the second and third trimesters were studied in 18 previously sedentary women (exercised group, EG). A nonexercising pregnant control group (CG, n = 9) was also studied. Data collection times for both groups were as follows: start of the second trimester (Entry), ends of the second (TM2) and third (TM3) trimesters (post-training), and 4-6 months postpartum (nonpregnant control). Respiratory gas exchange was studied and venous blood samples were obtained before, during, and after a graded cycle ergometer test that was terminated at a peak heart rate of 170 beats/min. Measurements included plasma glucose, insulin, free fatty acids, the respiratory exchange ratio at peak exercise, and peak postexercise lactate concentration. A significant aerobic conditioning effect in the EG was confirmed by a 17% increase in O2 pulse at peak exercise between Entry and TM3. As expected, values for free fatty acids in the CG rose with advancing gestational age. The CG showed a clear trend for a rise in plasma insulin with advancing gestational age, under all experimental conditions. Also, peak exercise respiratory exchange ratio and peak postexercise lactate concentration were significantly reduced in late gestation, and plasma glucose decreased significantly during and following the end of TM3 testing. Effects of pregnancy to reduce peak postexercise lactate and to reduce plasma glucose during and after exercise at the end of the third trimester were significantly attenuated in the EG. These effects were attributed to attenuation of pregnancy-induced insulin resistance (as reflected by insulin/glucose ratio) by physical conditioning. These findings support our original experimental hypothesis that aerobic conditioning prevents exercise-induced hypoglycemia and preserves the ability to utilize carbohydrate and produce lactate during heavy exercise in late gestation. PMID- 12897818 TI - Vascular responses to alpha-adrenergic stimulation and depolarization are enhanced in insulin-resistant and diabetic Psammomys obesus. AB - Since vascular complications often accompany diabetes, we examined the influence of the endothelial lining on vascular reactivity in Psammomys obesus, a desert gerbil that acquires insulin resistance and diabetes when exposed to a laboratory diet. Vasoconstriction to phenylephrine and depolarizing KCl, as well as carbachol endothelium-dependent relaxation, were assessed in rings of thoracic aortae obtained from three groups: (i) group A, normoglycemic-normoinsulinemic; (ii) group B, normoglycemic-hyperinsulinemic, and (iii) group C, hyperglycemic hyperinsulinemic animals. As expected, marked hypertriglyceridemia and hypercholesterolemia characterized groups B and C, which developed enhanced contractile responsiveness to phenylephrine and KCl compared with controls (group A). Furthermore, both experimental groups displayed a significant decrease in endothelium-dependent relaxation to carbachol. Altered lipid profiles are considered to play some role in the observed modification of aortic reactivity. Overall, our data indicate that vascular contractile responsiveness is enhanced early in the development of insulin resistance and diabetes in the female P. obesus. PMID- 12897819 TI - Chronic carbon monoxide exposure of hypoxic rats increases in vitro sensitivity of pulmonary artery smooth muscle. AB - Exogenous carbon monoxide (CO) induces pulmonary vasodilation by acting directly on pulmonary artery (PA) smooth muscle cells. We investigated the contribution of K+ channels and soluble guanylyl cyclase to the regulation of PA tone by acute CO in chronic hypoxic rats (3 weeks at 0.5 atm (1 atm = 101.325 kPa); hypoxic) and in chronic hypoxic rats exposed to exogenous CO (3 weeks at 0.5 atm + 50 ppm CO; hypoxic-CO). Acute CO induced relaxation in PA rings from all animals. However, the amplitude of CO relaxation was significantly decreased in hypoxic rings and increased in hypoxic-CO rings. This different effect occurred with a decrease and an increase of pD2, respectively, in hypoxic and hypoxic-CO rings. We showed a positive relation between the percentage of inhibition of CO relaxation by a blocker of K+ channels and the increase of CO sensitivity. Thus, we showed for the first time that chronic hypoxia decreases acute CO sensitivity, which in contrast, increases in the presence of chronic CO. The present study provides initial evidence of a link between increased K(+)-channel activity and CO sensitivity. PMID- 12897820 TI - Effects of PACAP(1-27) on the canine endocrine pancreas in vivo: interaction with cholinergic mechanism. AB - The aim of the present study was to characterize the effects of pituitary adenylate cyclase activating polypeptide (PACAP) on the endocrine pancreas in anesthetized dogs. PACAP(1-27) and a PACAP receptor (PAC(1)) blocker, PACAP(6 27), were locally administered to the pancreas. PACAP(1-27) (0.005-5 microg) increased basal insulin and glucagon secretion in a dose-dependent manner. PACAP(6-27) (200 microg) blocked the glucagon response to PACAP(1-27) (0.5 microg) by about 80%, while the insulin response remained unchanged. With a higher dose of PACAP(6-27) (500 microg), both responses to PACAP(1-27) were inhibited by more than 80%. In the presence of atropine with an equivalent dose (128.2 microg) of PACAP(6-27) (500 microg) on a molar basis, the insulin response to PACAP(1-27) was diminished by about 20%, while the glucagon response was enhanced by about 80%. The PACAP(1-27)-induced increase in pancreatic venous blood flow was blocked by PACAP(6-27) but not by atropine. The study suggests that the endocrine secretagogue effect of PACAP(1-27) is primarily mediated by the PAC(1) receptor, and that PACAP(1-27) may interact with muscarinic receptor function in PACAP-induced insulin and glucagon secretion in the canine pancreas in vivo. PMID- 12897821 TI - Ethyl isopropyl amiloride inhibits smooth muscle cell proliferation and migration by inducing apoptosis and antagonizing urokinase plasminogen activator activity. AB - Amiloride inhibits activation of the Na(+)-H+ exchanger (NHE), a critical step in smooth muscle cell (SMC) growth. While amiloride treatment reduces SMC proliferation and migration, as well as experimental lesion formation, these effects are not exclusively due to NHE inhibition and remain incompletely understood. The purpose of this study was to examine the mechanisms involved in amiloride-induced attenuation of SMC proliferation and migration, looking specifically at the potential role of apoptosis and urokinase plasminogen activator (uPA) activity in these processes. Rabbit SMCs in tissue culture were exposed to 10-80 microM of the amiloride analogue ethyl isopropyl amiloride (EIPA). Compared with controls, EIPA reduced DNA synthesis, cell number, and mitochondrial respiration, but without toxic effects on quiescent or proliferating cells. In a Boyden chamber assay, EIPA reduced uPA-induced SMC migration. Moreover, in a SMC scratch assay EIPA treatment resulted in a 66% reduction in the number of repopulating cells, a 92% decrease in the number of proliferating cells, and a 37-fold increase in the number of apoptotic cells. SMC apoptosis was frequently localized to the scratch edges, where cell proliferation and bcl-2 expression were absent. Finally, uPA enzymatic activity in the cell culture media was lower for EIPA-treated versus control SMCs. Therefore, EIPA inhibits both SMC proliferation and migration by inducing apoptosis and antagonizing uPA activity, respectively, and requires further study as an agent for reducing vascular lesion formation. PMID- 12897822 TI - Bradykinin metabolism in rat hearts with left-ventricular hypertrophy following myocardial infarction. AB - The aim of the present study was to assess the contribution of angiotensin I converting enzyme (ACE)and neutral endopeptidase (NEP) in the coronary degradation of bradykinin (BK) after left-ventricular hypertrophy following myocardial infarction (MI) in rats. Myocardial infarction was induced by left descendant coronary artery ligation, and the contribution of ACE and NEP in the degradation of exogenous BK after a single passage through the coronary bed was assessed at 2, 5, and 36 days post-MI. BK degradation rate (V(max)/Km) was found to be significantly lower in hearts at 36 days (3.30 +/- 0.28 min(-1)) compared with 2 days (4.39 +/- 0.32 min(-1)) for noninfarcted hearts, but this reduction was just above the statistical level of significance for post-MI hearts. In infarcted hearts, V(max)/Km was increased significantly 5 days post-MI (4.91 +/- 0.28 min(-1)) compared with the 2 and 36 day-groups (3.43 +/- 0.20 and 2.78 +/- 0.16 min(-1), respectively). The difference between noninfarcted and MI was significant only 2 days post-MI. Treatment with the vasopeptidase inhibitor, omapatrilat, showed that the relative contribution of ACE and NEP combined increased over time in infarcted hearts and became significantly higher 36 versus 2 days post-MI. Finally, the treatment with an ACE inhibitor (enalaprilat) and a NEP inhibitor (retrothiorphan) in the 36-day infarcted and noninfarcted hearts showed that the relative contribution of ACE in infarcted hearts was comparable with that of noninfarcted hearts, whereas the relative contribution of NEP was increased significantly in infarcted hearts. In conclusion, experimental MI in rats induces complex changes in the metabolism of exogenous BK. The changes resulted in an increased relative contribution of NEP 36 days after infarction. PMID- 12897823 TI - Serotonin effect on deiodinating activity in the rat. AB - Serotonin (5-HT) and thyroid hormones are part of a complex system modulating eating behaviour and energy expenditure. 5-Deiodinase (5-D) converts the relatively inactive thyroxine (T4) to triiodothyronine (T3), and its activity is an indirect measure of T3 production in peripheral tissues, particularly in the brain, intrascapular brown adipose tissue (IBAT), heart, liver, and kidney. We evaluated the effect of 5-HT on 5'-D activity during basal conditions and after short (30 min) cold exposure (thyroid stimulating hormone stimulation test, TST). 5'-D activity was assessed in the liver, heart, brain, kidney, and IBAT. TST increases 5'-D activity in the brain, heart, and IBAT and decreases it in kidney, leaving it unchanged in the liver. 5-HT alone did not modify 5'-D activity in the organs under study but decreased it in the IBAT, heart, and brain when injected before the TST was administered. Our results confirm the important role of 5-HT in thermoregulation, given its peripheral site of action, in modulating heat production controlling intracellular T3 production. These effects are more evident when heat production is upregulated during cold exposure in organs containing type II 5'-D, such as the brain, heart, and IBAT, which are able to modify their function during conditions that alter energy balance. In conclusion, 5-HT may also act peripherally directly on the thyroid and organs containing type II 5'-D, thus controlling energy expenditure through heat production. PMID- 12897824 TI - Bacterial extracellular protease activities in field soils under different fertilizer managements. AB - The major extracellular endopeptidase from Bacillus subtilis PF212 (isolated from paddy field soil) and B. subtilis CF80 (isolated from upland field soil) belongs to the group of serine proteases produced by Bacillus spp. known as subtilisins (optimum pH 7.0, optimum temperature 60 degrees C, and molecular mass 28 kDa). The NH2-terminal amino acid sequence (20 amino acids) of the endopeptidase from (i) strain CF80 was identical with that of subtilisin BPN' and (ii) strain PF212 was identical with that of subtilisin Amylosacchariticus. The properties (i.e., effect of inhibitors) of these endopeptidases were similar to those of the overall soil endopeptidase and soil endopeptidases extracted from paddy field soil. From the numbers of B. subtilis we isolated from paddy fields and found to produce a subtilisin-like serine protease, it seemed possible to consider that subtilisin was one of the soil endopeptidases in paddy field soils. The major extracellular endopeptidase from Serratia marcescens (strains 4-12-132, 4-12-131, and 4-60-110) isolated from upland field soils applied with animal slurry is a serratial metalloprotease (optimum pH 9.5, optimum temperature 40 degrees C, and molecular mass 50 kDa). The NH2-terminal amino acid sequence (20 amino acids) of the endopeptidase from strain 4-12-132 was identical with that of serratial metalloprotease, and partial DNA sequence of the endopeptidase gene of S. marcescens 4-12-132 had high homology with that of the serratial metalloprotease gene. The properties (i.e., effect of inhibitors) of this endopeptidase were similar to those of the overall soil endopeptidase in upland fields applied with animal slurry. Thus, it was possible to consider that serratial metalloprotease was one of the soil endopeptidases in upland fields applied with animal slurry. PMID- 12897825 TI - Chromosome DNA fragmentation and excretion caused by defective prophage gene expression in the early-exponential-phase culture of Bacillus subtilis. AB - Bacillus subtilis 168 and its major autolysin mutant, AN8, were shown to excrete two size classes of DNA when cultured in Luria-Bertani medium. Pulsed-field gel electrophoresis of DNA harvested from the cell surface demonstrated the presence of 13-kb-long and circa 50-kb-long strands. Restriction digestion of both sizes of DNA resulted in a smearing pattern, as observed by agarose gel electrophoresis. Shotgun sequencing of DNase I partial digests of 50-kb DNA fragments revealed that the strands originate from various sites on the chromosome. SDS-PAGE analysis of cell surface fractions and culture supernatants demonstrated the presence of several proteins that were thought to be associated with the DNA. Of these, three major proteins were identified, i.e., XkdG, XkdK, and XkdM, by tandem mass spectrometry, all of which were proteins of a defective prophage PBSX residing in the Bacillus subtilis chromosome. Disruption of these PBSX genes resulted in a reduction of 13-kb fragment generation and excretion and also a great reduction of 50-kb fragment excretion. Electron microscopy showed that a few mature phages and numerous membrane vesicle-like particles existed in the cell surface fractions of strain 168. The present findings suggest that the spontaneous generation and excretion of chromosome DNA fragments in Bacillus subtilis are both closely related to the expression of defective prophage genes. PMID- 12897826 TI - Prevalence of Escherichia coli O157:H7 and Salmonella spp. in surface waters of southern Alberta and its relation to manure sources. AB - The Oldman River watershed in southern Alberta, Canada, is an extensively irrigated region in which intensive agricultural practices have flourished. Concern over water quality in the basin has been expressed because of high levels of enteric disease indigenous to the region. To address these concerns, we conducted a 2-year study to estimate the prevalence of Escherichia coli O157:H7 and Salmonella spp. in surface water within the basin. This study is the first of its kind to identify E. coli O157:H7 repeatedly in surface water collected from a Canadian watershed. Prevalence of E. coli O157:H7 and Salmonella spp. in water samples was 0.9% (n = 1,483) and 6.2% (n = 1,429), respectively. While data examined at a regional level show a relationship between high livestock density and high pathogen levels in southern Alberta, statistical analysis of point source data indicates that predicted manure output from bovine, swine, and poultry feeding operations was not directly associated with either Salmonella spp. or E. coli O157:H7 prevalence. However, geography and weather variables, which are likely to influence bacterial runoff, were not considered in this model. We also postulate that variations in time, amount, and frequency of manure application onto agricultural lands may have influenced levels of surface-water contamination with these bacterial pathogens. PMID- 12897827 TI - Toxic effects caused by heavy metals in the yeast Saccharomyces cerevisiae: a comparative study. AB - The decreasing order of toxicity of select heavy metals on the yeast Saccharomyces cerevisiae, in 10 mM MES (2-(N-morpholino)ethanesulfonic acid) pH buffer at pH 6.0, was found to be copper, lead, and nickel. Heavy metal (200 microM) induced a decrease in the number of viable cells by about 50% in the first 5 min for copper and in 4 h for lead, while nickel was not toxic up to a 200 microM concentration over a period of 48 h. Glucose (25 mM) strongly enhanced the toxic effect of 50 microM copper but had little or no effect on the toxicity of 200 microM lead or nickel. Copper, lead, and nickel induced the leakage of UV260-absorbing compounds from cells with different kinetics. The addition of 0.5 mM calcium, before addition of 200 microM copper, showed a protective action against cell death and decreased the release of UV-absorbing compounds, while no effect was observed against lead or nickel toxic effects. Copper complexation capacities of the filtrates of cells exposed for 2 h in 200 microM copper and 24 h in 200 microM lead were 51 and 14 microM, respectively. The implication of the complexation shown by these soluble compounds in the bioavailability of heavy metals is discussed. PMID- 12897828 TI - Modulatory effect of cAMP on fungal ergosterol level and inhibitory activity of azole drugs. AB - The functions and biosynthesis of sterols have been effective targets for fungal control in different areas, including pharmaceutical and agricultural applications. Fungi are among the organisms that synthesize sterols, principally ergosterol. In this paper, the effect of dibutyryl-cAMP (db-cAMP) on ergosterol level and the interaction of drugs that would change the concentration of cAMP with antifungal drugs have been investigated. Sterols were extracted from Candida albicans, and ergosterol was measured using the gas chromatography method. The interaction of different agents was measured by the broth dilution method. It was found that phosphodiesterase inhibitors reverse the inhibitory activity of azole antifungal drugs. Evaluating the ergosterol level of C. albicans incubated with db-cAMP revealed that it increased ergosterol level. Further experiments provided evidence attributing the observed interaction between azoles and phosphodiesterase inhibitors to the relationship between ergosterol and cAMP. The possible significance of this interaction includes potentiation of antifungal activity of drugs by manipulating the cAMP level. PMID- 12897829 TI - Characterization of group B streptococcal glyceraldehyde-3-phosphate dehydrogenase: surface localization, enzymatic activity, and protein-protein interactions. AB - During characterization of the surface antigens of serotype III group B streptococci (GBS), a protein with an apparent M(r) of approximately 173,500 migrating on a SDS--polyacrylamide gel was found to have an N-terminal amino acid sequence identical to that of the plasmin receptor (Plr) of group A streptococci, a surface-localized glyceraldehyde-3-phosphate dehydrogenase (GAPDH). This work begins to characterize GBS GAPDH and to assess its functional activity on the cell surface. The 1.0-kb gapC gene of GBS was amplified by PCR. plr and gapC demonstrated 87% homology. An anti-Plr monoclonal antibody reacted with GBS whole cells, suggesting GBS GAPDH is surface localized. Multiple serotypes of GBS demonstrated functional GAPDH on their surfaces. The anti-Plr monoclonal antibody recognized GBS protein bands of approximately 41 and 173.5 kDa, by Western blot. Presumably, these represent monomeric and tetrameric forms of the GAPDH molecule. GBS GAPDH was demonstrated by Western blot analysis to interact with lys- and glu plasminogens. Fluid-phase GBS GAPDH interacted, by means of ELISA, with immobilized lys-plasminogen, glu-plasminogen, actin, and fibrinogen. Enzymatically active GAPDH, capable of binding cytoskeletal and extracellular matrix proteins, is expressed on the surface of GBS. PMID- 12897830 TI - Wild-type Escherichia coli producing microcins B17, D93, J25, and L; cloning of genes for microcin L production and immunity. AB - For the first time, an Escherichia coli strain producing four microcins (Mcc), B17, D93, J25, and L, and showing immunity to Mcc V was isolated and characterized. Each of the gene clusters encoding the production of Mcc B17, D93, and L was cloned separately. The gene cluster for Mcc L was cloned within a 13.5 kb HindIII-SalI fragment, which includes the Mcc V immunity gene, cvi. PMID- 12897831 TI - High TNT-transforming activity by a mixed culture acclimated and maintained on crude-oil-containing media. AB - A mixed microbial culture originating from a petroleum-contaminated site and maintained on crude oil exhibited high 2,4,6-trinitrotoluene (TNT) transformation activity. Cultivation of the mixed culture in glucose-containing medium for 29 h resulted in almost complete transformation of 100 ppm TNT. TNT transformation was observed with both growing and resting cells. With subculturing, it was found that TNT could support growth of the mixed culture when supplied as sole carbon source, sole nitrogen source, or sole carbon and nitrogen source. The finding that a mixed microbial culture maintained on crude oil exhibited high TNT transformation activity without prior subculture on TNT-containing media is novel and may have potential practical applications in the bioremediation of munitions contaminated soil and wastewater. PMID- 12897832 TI - Comparative analysis of the Bradyrhizobium japonicum sucA region. AB - To study the adjustments made to the tricarboxylic acid cycle during symbiosis of nitrogen-fixing rhizobia with their host legumes, we have characterized the genes encoding the alpha-ketoglutarate dehydrogenase enzyme complex in Bradyrhizobium japonicum. The genes were arranged in the order sucA-sucB-scdA-lpdA, where scdArepresents a short-chain dehydrogenase gene (GenBank accession No. AY049030). All four genes appeared to be co-transcribed, an arrangement that is so far unique to B. japonicum. The mdh gene, encoding malate dehydrogenase, was located upstream of the sucA operon, and its primary transcript appeared to be monocistronic. Primer extension indicated that the sucA operon and mdh were transcribed from typical housekeeping promoters. PMID- 12897833 TI - Marinobacter lutaoensis sp. nov., a thermotolerant marine bacterium isolated from a coastal hot spring in Lutao, Taiwan. AB - A heterotrophic and thermotolerant marine bacterium, designated strain T5054, was isolated from a hot spring on the coast of Lutao, Taiwan. It was a strictly aerobic, Gram-negative rod. Cells grown in broth cultures were non-spore-forming and motile by means of one to several polar flagella. It seems that pilus-like structures were produced from both poles of the cells. Strain T5054 required Na+ for growth and exhibited optimal growth at about 45 degrees C, pH 7, and 3-5% NaCl. It contained iso-C15:0 as the most abundant fatty acid and ubiquinone-8 as the only isoprenoid quinone. Its genomic DNA G+C content was 63.5 mol%. The strain did not require vitamins or other organic growth factors, and it grew on glucose, mannitol, and a variety of organic acids and amino acids as sole carbon sources. Characterization data, together with the results of a 16S rDNA-based phylogenetic analysis, indicated that strain T5054 could be classified as a new species in the genus Marinobacter. The name Marinobacter lutaoensis sp. nov. is proposed for this new bacterium. The type strain is T5054 (CCRC 17087; JCM 11179). PMID- 12897834 TI - Characterization of an antifungal soil bacterium and its antagonistic activities against Fusarium species. AB - Bacteria were isolated from a cultivated soil and screened for antagonistic activity against Fusarium graminearum, a predominant agent of ear rot and head blight in cereal crops. Based on its in vitro effectiveness, isolate D1/2 was selected for characterization and identified as a strain of Bacillus subtilis by phenotypic tests and comparative analysis of its 16S ribosomal RNA gene (rDNA) sequence. It inhibited the mycelial growth of a collection of common fungal phytopathogens, including eight Fusarium species, three other ascomycetes, and one basidiomycete. The cell-free culture filtrate of D1/2 at different dilutions was active against macroconidium germination and hyphal growth of F. graminearum, depending on the initial macroconidium density. It induced the formation of swollen hyphal cells in liquid cultures of this fungus grown from macroconidia. A bioassay also demonstrated that D1/2 offered in planta protection against the damping-off disease in alfalfa seedlings caused by F. graminearum, while the type strain of B. subtilis was ineffective. Hence, B. subtilis D1/2 or its culture filtrate has potential application in controlling plant diseases caused by Fusarium. PMID- 12897835 TI - Release and regeneration of protoplasts from the fungus Trichothecium roseum. AB - A protocol for isolating and regenerating protoplasts from Trichothecium roseum has been described. Protoplasts from T. roseum were isolated using (i) a lytic enzyme combination composed of Novozym 234, chitinase, cellulase, and pectinase at a 5-mg/mL concentration and (ii) 0.6 M KCl as an osmotic stabilizer. A maximum number of 28 x 10(4) protoplasts/mL were obtained at pH 5.5. Experiments on the regeneration and reversion of protoplasts revealed a maximum regeneration (60.8%) in complete medium (potato dextrose--yeast extract agar) amended with 0.6 M KCl. The regenerated protoplasts were similar to the original parent strain in morphology, pigmentation, growth, and sporulation. PMID- 12897836 TI - A phylogenetic analysis of the pSymB replicon from the Sinorhizobium meliloti genome reveals a complex evolutionary history. AB - Microbial genomes are thought to be mosaic, making it difficult to decipher how these genomes have evolved. Whole-genome nearest-neighbor analysis was applied to the Sinorhizobium meliloti pSymB replicon to determine its origin, the degree of horizontal transfer, and the conservation of gene order. Prediction of the nearest neighbor based on contextual information, i.e., the nearest phylogenetic neighbor of adjacent genes, provided useful information for genes for which phylogenetic relationships could not be established. A large portion of pSymB genes are most closely related to genes in the Agrobacterium tumefaciens linear chromosome, including the rep and min genes. This suggests a common origin for these replicons. Genes with the nearest neighbor from the same species tend to be grouped in "patches". Gene order within these patches is conserved, but the content of the patches is not limited to operons. These data show that 13% of pSymB genes have nearest neighbors in species that are not members of the Rhizobiaceae family (including two archaea), and that these likely represent genes that have been involved in horizontal transfer. PMID- 12897837 TI - Relative importance of the endomycorrhizal and (or) ectomycorrhizal associations in Allocasuarina and Casuarina genera. AB - This work was carried out to determine the relative importance of the endomycorrhizal and (or) ectomycorrhizal association in species of Casuarina and Allocasuarina. Under axenic conditions, Pisolithus and Scleroderma isolates formed ectomycorrhizas with a mantle and a Hartig net on Allocasuarina verticillata but failed to form a Hartig net on Casuarina glauca. In a controlled soil system, C. glauca was inoculated with the endomycorrhizal fungus Glomus intraradices Schenck & Smith, and A. verticillata was inoculated with Pisolithus albus IR100 Bougher & Smith and (or) G. intraradices. Both symbionts significantly stimulated growth in both plant species. For A. verticillata, its growth response to ectomycorrhizal inoculation was higher than to endomycorrhizal inoculation. When both symbionts were inoculated, antagonism among the fungal isolates was observed with a higher ectomycorrhizal colonization. These results showed that A. verticillata was ectomycorrhizal dependent, whereas C. glauca was endomycorrhizal dependent. From a practical point of view, this study shows the importance of selecting compatible mycorrhizal fungi for developing successful inoculation programmes. In addition, it would help to further research and determine the effect of ecto- and endo-mycorrhizal symbiosis on the formation and function of N2-fixing actinorhizal nodules. PMID- 12897838 TI - Release of Ni and Co by microbial activity in New Caledonian ultramafic soils. AB - A high positive correlation was found between extractable Ni and Co contents and microbial activity of 40 ultramafic soil samples from New Caledonia, suggesting a possible role of microorganisms in the release of these two metals. A saprolite (ultramafic subsoil) and a hypermagnesian brown soil were incubated 9 months in different conditions. Ni and Co release, measured by their extractability, occurred without reduction of soil pH but did not occur when the native microflora was eliminated by heat treatment. However, when autoclaved soil was re inoculated with a pinch of the same nonheated soil, the release of metals was recorded. The concentrations of extractable Ni and Co were much higher in soils amended with organic compounds in which microbial activity was enhanced, but only if the soil was not heat treated. The presence of Grevillea exul, a metal tolerant plant, stimulated the metal release process, but the stimulation was less effective than it was in the compost-amended soil without plant. The influence of the plant in extractable Ni and Co contents in this amended soil was not significant. The release of the two metals therefore seemed to be induced by the activity of specialized organotrophic microorganisms. PMID- 12897839 TI - Characterization of the sodF gene region of Frankia sp. strain ACN14a and complementation of Escherichia coli sod mutant. AB - The Frankia sp. strain ACN14a superoxide dismutase SodF was previously shown to be induced in response to Alnus glutinosa root exudates, and its gene was sequenced. We report here the sequence of the 9-kb genomic segment surrounding the sodF gene and further characterize this gene and its product. Nine ORFs coding for various proteins, such as regulators, acetyl-CoA transferases, and a bacterioferritin A next to the sodF gene, were found. Northern blot analysis showed that the sodF gene was expressed as a major 1-kb transcript, which indicates that it has its own promoter. The sodF gene strongly complemented an Escherichia coli triple mutant (sodA sodB recA), restoring aerobic growth when the gene was expressed from the synthetic tac promoter but when expressed from its own promoter showed only slight rescue, suggesting that it was poorly recognized by the E. coli RNA polymerase. It is noteworthy that this is the first time that a Frankia gene has been reported to complement an E. coli mutant. The superoxide dismutase activity of the protein was inactivated by hydrogen peroxide, indicating that the metal ligand is iron, which is supported by analysis of the protein sequence. Thus, the SodF protein induced in Frankia by root exudates is an iron-containing enzyme similar to the one present in the nodules. PMID- 12897840 TI - Transforming the sapstaining fungus Ophiostoma piceae with Agrobacterium tumefaciens. AB - A transformation protocol mediated by Agrobacterium tumefaciens is described for the sapstaining fungus Ophiostoma piceae. We compared transformants obtained from Agrobacterium with those obtained from yeast-like cells made into spheroplasts and treated with CaCl2. For all putative transformants analyzed, Southern hybridization confirmed that the hygromycin resistance gene had been integrated into the genomic DNA. While all transformants obtained from the treated spheroplasts had multiple copy vector insertion, 85% of the Agrobacterium mediated transformants had single copy vector insertion. PMID- 12897842 TI - High concentration of DNA in condensed chromatin. AB - The lengths of the DNA molecules of eukaryotic genomes are much greater than the dimensions of the metaphase chromosomes in which they are contained during mitosis. From this observation it has been generally assumed that the linear packing ratio of DNA is an adequate measure of the degree of DNA compaction. This review summarizes the evidence suggesting that the local concentration of DNA is more appropriate than the linear packing ratio for the study of chromatin condensation. The DNA concentrations corresponding to most of the models proposed for the 30-40 nm chromatin fiber are not high enough for the construction of metaphase chromosomes. The interdigitated solenoid model has a higher density because of the stacking of nucleosomes in secondary helices and, after further folding into chromatids, it yields a final concentration of DNA that approaches the experimental value found for condensed chromosomes. Since recent results have shown that metaphase chromosomes contain high concentrations of the chromatin packing ions Mg2+ and Ca2+, it is discussed that dynamic rather than rigid models are required to explain the condensation of the extended fibers observed in the absence of these cations. Finally, considering the different lines of evidence demonstrating the stacking of nucleosomes in different chromatin complexes, it is suggested that the face-to-face interactions between nucleosomes may be the driving force for the formation of higher order structures with a high local concentration of DNA. PMID- 12897843 TI - Getting into chromatin: how do transcription factors get past the histones? AB - Transcriptional activators and the general transcription machinery must gain access to DNA that in eukaryotes may be packaged into nucleosomes. In this review, I discuss this problem from the standpoint of the types of chromatin structures that these DNA-binding proteins may encounter, and the mechanisms by which they may contend with various chromatin structures. The discussion includes consideration of experiments in which chromatin structure is manipulated in vivo to confront activators with nucleosomal binding sites, and the roles of nucleosome dynamics and activation domains in facilitating access to such sites. Finally, the role of activators in facilitating access of the general transcriptional machinery to sites in chromatin is discussed. PMID- 12897844 TI - HMGN dynamics and chromatin function. AB - Recent studies indicate that most nuclear proteins, including histone H1 and HMG are highly mobile and their interaction with chromatin is transient. These findings suggest that the structure of chromatin is dynamic and the protein composition at any particular chromatin site is not fixed. Here we discuss how the dynamic behavior of the nucleosome binding HMGN proteins affects the structure and function of chromatin. The high intranuclear mobility of HMGN insures adequate supply of protein throughout the nucleus and serves to target these proteins to their binding sites. Transient interactions of the proteins with nucleosomes destabilize the higher order chromatin, enhance the access to nucleosomal DNA, and impart flexibility to the chromatin fiber. While roaming the nucleus, the HMGN proteins encounter binding partners and form metastable multiprotein complexes, which modulate their chromatin interactions. Studies with HMGN proteins underscore the important role of protein dynamics in chromatin function. PMID- 12897845 TI - Characteristics of gamma-H2AX foci at DNA double-strand breaks sites. AB - Phosphorylated H2AX (gamma-H2AX) is essential to the efficient recognition and (or) repair of DNA double strand breaks (DSBs), and many molecules, often thousands, of H2AX become rapidly phosphorylated at the site of each nascent DSB. An antibody to gamma-H2AX reveals that this highly amplified process generates nuclear foci. The phosphorylation site is a serine four residues from the C terminus which has been evolutionarily conserved in organisms from giardia intestinalis to humans. Mice and yeast lacking the conserved serine residue demonstrate a variety of defects in DNA DSB processing. H2AX Delta/Delta mice are smaller, sensitive to ionizing radiation, defective in class switch recombination and spermatogenesis while cells from the mice demonstrate substantially increased numbers of genomic defects. gamma-H2AX foci formation is a sensitive biological dosimeter and presents new and exciting opportunities to understand important biological processes, human diseases, and individual variations in radiation sensitivity. These potentialities demonstrate the importance of understanding the parameters and functions of gamma-H2AX formation. PMID- 12897846 TI - A haploid affair: core histone transitions during spermatogenesis. AB - The process of meiosis reduces a diploid cell to four haploid gametes and is accompanied by extensive recombination. Thus, the dynamics of chromatin during meiosis are significantly different than in mitotic cells. As spermatogenesis progresses, there is a widespread reorganization of the haploid genome followed by extensive DNA compaction. It has become increasingly clear that the dynamic composition of chromatin plays a critical role in the activities of enzymes and processes that act upon it. Therefore, an analysis of the role of histone variants and modifications in these processes may shed light upon the mechanisms involved and the control of chromatin structure in general. Histone variants such as histone H3.3, H2AX, and macroH2A appear to play key roles in the various stages of spermiogenesis, in addition to the specifically modulated acetylation of histone H4 (acH4), ubiquitination of histones H2A and H2B (uH2A, uH2B), and phosphorylation of histone H3 (H3p). This review will examine recent discoveries concerning the role of histone modifications and variants during meiosis and spermatogenesis. PMID- 12897847 TI - The complex interactions of p53 with target DNA: we learn as we go. AB - The most import biological function of the tumor suppressor p53 is that of a sequence-specific transactivator. In response to a variety of cellular stress stimuli, p53 induces the transcription of an ever-increasing number of target genes, leading to growth arrest and repair, or to apoptosis. Long considered as a "latent" DNA binder that requires prior activation by C-terminal modification, recent data provide strong evidence that the DNA binding activity of p53 is strongly dependent on structural features within the target DNA and is latent only if the target DNA lacks a certain structural signal code. In this review we discuss evidence for complex interactions of p53 with DNA, which are strongly dependent on the dynamics of DNA structure, especially in the context of chromatin. We provide a model of how this complexity may serve to achieve selectivity of target gene regulation by p53 and how DNA structure in the context of chromatin may serve to modulate p53 functions. PMID- 12897848 TI - Magnetic tweezers: a sensitive tool to study DNA and chromatin at the single molecule level. AB - The advent of single-molecule biology has allowed unprecedented insight into the dynamic behavior of biological macromolecules and their complexes. Unexpected properties, masked by the asynchronous behavior of myriads of molecules in bulk experiments, can be revealed; equally importantly, individual members of a molecular population often exhibit distinct features in their properties. Finally, the single-molecule approaches allow us to study the behavior of biological macromolecules under applied tension or torsion; understanding the mechanical properties of these molecules helps us understand how they function in the cell. In this review, we summarize the application of magnetic tweezers (MT) to the study of DNA behavior at the single-molecule level. MT can be conveniently used to stretch DNA and introduce controlled levels of superhelicity into the molecule and to follow to a high definition the action of different types of topoisomerases. Its potential for chromatin studies is also enormous, and we will briefly present our first chromatin results. PMID- 12897849 TI - The many roles of the transcriptional regulator CTCF. AB - The nuclear factor CTCF was first identified as one of the factors binding to the regulatory regions of the c-myc gene. Further study of this protein revealed roles in transcriptional repression, insulator function, and imprinting genetic information. Recent studies have provided new insight into the mechanism through which this factor acts at various levels of gene regulation. PMID- 12897850 TI - Models for chromatin remodeling: a critical comparison. AB - Nucleosome remodeling has been shown, in many cases, to involve cis displacement of nucleosomes on the DNA. This process seems similar to the long-recognized random diffusion of nucleosomes along DNA, but the remodeling process is unidirectional and ATP dependent. Several years ago, we developed a model for nucleosome migration, based on the diffusion of "twist-defects" within the nucleosomal DNA. This has been modified into a model that incorporates ATP dependent defect generation, and can account for many observations concerning remodeling. However, certain experimental studies in recent years have cast doubt on the applicability of the twist-diffusion model for remodeling, and seem to favor instead a "reptation" model. We discuss herein these problems and propose a resolution. PMID- 12897851 TI - Revisiting the structure and functions of the linker histone C-terminal tail domain. AB - Linker histones stabilize folded chromatin, acting through their long C-terminal tails. The C-termini contain high percentages of evenly distributed lysine and arginine residues and have no secondary structure in solution. Hence, it has generally been believed that the C-termini function by shielding negative charges on the DNA backbone. However, recent evidence supports a mechanism of action of the linker histone C-terminus that involves formation of specific secondary structure(s) upon interaction with other components of the chromatin fiber. PMID- 12897852 TI - Xenopus transcription factor IIIA and the 5S nucleosome: development of a useful in vitro system. AB - 5S RNA genes in Xenopus are regulated during development via a complex interplay between assembly of repressive chromatin structures and productive transcription complexes. Interestingly, 5S genes have been found to harbor powerful nucleosome positioning elements and therefore have become an important model system for reconstitution of eukaryotic genes into nucleosomes in vitro. Moreover, the structure of the primary factor initiating transcription of 5S DNA, transcription factor IIIA, has been extensively characterized. This has allowed for numerous studies of the effect of nucleosome assembly and histone modifications on the DNA binding activity of a transcription factor in vitro. For example, linker histones bind 5S nucleosomes and repress TFIIIA binding in vitro in a similar manner to that observed in vivo. In addition, TFIIIA binding to nucleosomes assembled with 5S DNA is stimulated by acetylation or removal of the core histone tail domains. Here we review the development of the Xenopus 5S in vitro system and discuss recent results highlighting new aspects of transcription factor - nucleosome interactions, PMID- 12897853 TI - HMGA proteins: flexibility finds a nuclear niche? AB - The mammalian HMGA family of chromatin proteins possesses an unusual constellation of physical, biochemical, and biological characteristics that distinguish them from other nuclear proteins. Principal among these is the fact that, unlike other proteins, they possess little detectable secondary structure prior to interactions with other macromolecules (DNA, RNA, proteins). Upon binding to substrates, however, the HMGA proteins undergo specific disordered-to ordered structural transitions and also induce alterations in the structure of the substrates themselves. Their intrinsic structural flexibility, combined with other features such as the control of their substrate interactions via complex patterns of in vivo biochemical modifications, allows the HMGA proteins to actively participate in a wide variety of nuclear activities including DNA replication, DNA repair, chromatin remodeling, control of gene transcription, and regulation of mRNA processing PMID- 12897854 TI - A role for poly(ADP-ribosyl)ation in DNA methylation. AB - The aberrant DNA methylation of promoter regions of housekeeping genes leads to gene silencing. Additional epigenetic events, such as histone methylation and acetylation, also play a very important role in the definitive repression of gene expression by DNA methylation. If the aberrant DNA methylation of promoter regions is the starting or the secondary event leading to the gene silencing is still debated. Mechanisms controlling DNA methylation patterns do exist although they have not been ultimately proven. Our data suggest that poly(ADP ribosyl)ation might be part of this control mechanism. Thus an additional epigenetic modification seems to be involved in maintaining tissue and cell-type methylation patterns that when formed during embryo development, have to be rigorously conserved in adult organisms. PMID- 12897855 TI - Micromechanics of chromatin and chromosomes. AB - The enzymes that transcribe, recombine, package, and duplicate the eukaryotic genome all are highly processive and capable of generating large forces. Understanding chromosome function therefore will require analysis of mechanics as well as biochemistry. Here we review development of new biophysical-biochemical techniques for studying the mechanical properties of isolated chromatin fibers and chromosomes. We also discuss microscopy-based experiments on cells that visualize chromosome structure and dynamics. Experiments on chromatin tell us about its flexibility and fluctuation, as well as quantifying the forces generated during chromatin assembly. Experiments on whole chromosomes provide insight into the higher-order organization of chromatin; for example, recent experiments have shown that the mitotic chromosome is held together by isolated chromatin-chromatin links and not a large, mechanically contiguous non-DNA "scaffold". PMID- 12897856 TI - Histone H1 and the dynamic regulation of chromatin function. AB - Eukaryotic DNA is organized in a complex structure called chromatin. Although a primary function of chromatin is compaction of DNA, this must done such that the underlying DNA is potentially accessible to factor-mediated regulatory responses. Chromatin structure clearly plays a dominant role in regulating much of eukaryotic transcription. The demonstration that reversible covalent modification of the core histones contribute to transcriptional activation and repression by altering chromatin structure and the identification of numerous ATP-dependent chromatin remodeling enzymes provide strong support for this view. Chromatin is much more dynamic than was previously thought and regulation of the dynamic properties of chromatin is a key aspect of gene regulation. This review will focus on recent attempts to elucidate the specific contribution of histone H1 to chromatin-mediated regulation of gene expression. PMID- 12897857 TI - Sperm chromatin remodelling and Wolbachia-induced cytoplasmic incompatibility in Drosophila. AB - Wolbachia pipientis is an obligate bacterial endosymbiont, which has successfully invaded approximately 20% of all insect species by manipulating their normal developmental patterns. Wolbachia-induced phenotypes include parthenogenesis, male killing, and, most notably, cytoplasmic incompatibility. In the future these phenotypes might be useful in controlling or modifying insect populations but this will depend on our understanding of the basic molecular processes underlying insect fertilization and development. Wolbachia-infected Drosophila simulans express high levels of cytoplasmic incompatibility in which the sperm nucleus is modified and does not form a normal male pronucleus when fertilizing eggs from uninfected females. The sperm modification is somehow rescued in eggs infected with the same strain of Wolbachia. Thus, D. simulans has become an excellent model organism for investigating the manner in which endosymbionts can alter reproductive programs in insect hosts. This paper reviews the current knowledge of Drosophila early development and particularly sperm function. Developmental mutations in Drosophila that are known to affect sperm function will also be discussed.incompatibility. PMID- 12897858 TI - Communication over a large distance: enhancers and insulators. AB - Enhancers are regulatory DNA sequences that can work over a large distance. Efficient enhancer action over a distance clearly requires special mechanisms for facilitating communication between the enhancer and its target. While the chromatin looping model can explain the majority of the observations, some recent experimental findings suggest that a chromatin scanning mechanism is used to establish the loop. These new findings help to understand the mechanism of action of the elements that can prevent enhancer-promoter communication (insulators). PMID- 12897859 TI - Proceedings of the 24th International West Coast Chromatin and Chromosomes Conference. December 12-21, 2002. Pacific Grove, California, USA. PMID- 12897860 TI - Analysis of genetic diversity in red clover (Trifolium pratense L.) breeding populations as revealed by RAPD genetic markers. AB - Red clover is an important forage legume species for temperate regions and very little is known about the genetic organization of its breeding populations. We used random amplified polymorphic DNA (RAPD) genetic markers to address the genetic diversity and the distribution of variation in 20 breeding populations and cultivars from Chile, Argentina, Uruguay, and Switzerland. Genetic distances were calculated for all possible pairwise combinations. A high level of polymorphism was found and the proportion of polymorphic loci across populations was 74.2%. A population derived from a non-certified seedlot displayed a higher proportion of polymorphic loci than its respective certified seedlot. Gene diversity values and population genetics parameters suggest that the populations analyzed are diverse. An analysis of molecular variance (AMOVA) revealed that the largest proportion of variation (80.4%) resides at the within population level. RAPD markers are a useful tool for red clover breeding programs. A dendrogram based on genetic distances divided the breeding populations analyzed into three distinct groups. The amount and partition of diversity observed can be of value in identifying the populations that parents of synthetic cultivars are derived from and to exploit the variation available in the populations analyzed. PMID- 12897861 TI - Genetics of drought tolerance during seed germination in tomato: inheritance and QTL mapping. AB - A BC1 population (N = 1000) of an F1 hybrid between a stress-sensitive Lycopersicon esculentum breeding line (NC84173; maternal and recurrent parent) and a germination stress-tolerant Lycopersicon pimpinellifolium accession (LA722) was evaluated for seed germination rate under drought stress (DS) (14% w/v polyethyleneglycol-8000, water potential approximately -680 kPa), and the most rapidly germinating seeds (first 3% to germinate) were selected. The 30 selected BC1 seedlings were grown to maturity and self pollinated to produce BC1S1 progeny seeds. Twenty of the 30 selected BC1S1 progeny families were evaluated for germination rate under DS and their average performance was compared with that of a "nonselected" BC1S1 population of the same cross. Results indicated that selection for rapid germination under DS significantly improved progeny germination rate under DS (selection gain = 19.6%), suggesting a realized heritability of 0.47 for rate of germination under DS in this population. The 30 selected BC1 plants were subjected to restriction fragment length polymorphism (RFLP) analysis, and marker allele frequencies for 119 RFLP markers which spanned 1153 cM of the 12 tomato chromosomes were determined. A distributional extreme marker analysis, which measures statistical differences in marker allele frequencies between a selected and a nonselected population, detected four quantitative trait loci (QTLs) for rate of germination under DS in this population. Of these, two QTLs, located on chromosomes 1 and 9, were contributed by the L. pimpinellifolium donor parent and had larger effects than the other two QTLs, located on chromosomes 8 and 12, which were contributed by the L. esculentum recurrent parent. A few BC1S1 families were identified with all or most of the identified QTLs and with germination rates comparable with that of LA722. These families should be useful for the development of germination drought tolerant tomato lines using marker-assisted selection (MAS). The overall results indicate that drought tolerance during seed germination in tomato is genetically controlled and potentially could be improved by directional phenotypic selection or MAS. PMID- 12897862 TI - QTL mapping of partial resistance in winter wheat to Stagonospora nodorum blotch. AB - Stagonospora nodorum blotch is an important foliar and glume disease in cereals. Inheritance of resistance in wheat appears to be quantitative. To date, breeding of partially resistant cultivars has been the only effective way to combat this pathogen. The partial resistance components, namely length of incubation period, disease severity, and length of latent period, were evaluated on a population of doubled haploids derived from a cross between the partially resistant Triticum aestivum 'Liwilla' and susceptible Triticum aestivum 'Begra'. Experiments were conducted in a controlled environment and the fifth leaf was examined. Molecular analyses were based on bulked segregant analyses using 240 microsatellite markers. Four QTLs were significantly associated with partial resistance components and were located on chromosomes 2B, 3B, 5B, and 5D. The percentage of phenotypic variance explained by a single QTL ranged from 14 to 21% for incubation period, from 16 to 37% for disease severity, and from 13 to 28% for latent period, PMID- 12897863 TI - Molecular mapping of novel genes controlling Fusarium head blight resistance and deoxynivalenol accumulation in spring wheat. AB - Fusarium head blight of wheat is an extremely damaging disease, causing severe losses in seed yield and quality. The objective of the current study was to examine and characterize alternate sources of resistance to Fusarium head blight (FHB). Ninety-one F1-derived doubled haploid lines from the cross Triticum aestivum 'Wuhan-1' x Triticum aestivum 'Maringa' were examined for disease reaction to Fusarium graminearum by single-floret injection in replicated greenhouse trials and by spray inoculation in replicated field trials. Field and greenhouse experiments were also used to collect agronomic and spike morphology characteristics. Seed samples from field plots were used for deoxynivalenol (DON) determination. A total of 328 polymorphic microsatellite loci were used to construct a genetic linkage map in this population and together these data were used to identify QTL controlling FHB resistance, accumulation of DON, and agronomic and spike morphology traits. The analysis identified QTL for different types of FHB resistance in four intervals on chromosomes 2DL, 3BS, and 4B. The QTLs on 4B and 3BS proximal to the centromere are novel and not reported elsewhere. QTL controlling accumulation of DON independent of FHB resistance were located on chromosomes 2DS and 5AS. Lines carrying FHB resistance alleles on 2DL and 3BS showed a 32% decrease in disease spread after single-floret injection. Lines carrying FHB resistance alleles on 3BS and 4B showed a 27% decrease from the mean in field infection. Finally, lines carrying favourable alleles on 3BS and 5AS, showed a 17% reduction in DON accumulation. The results support a polygenic and quantitative mode of inheritance and report novel FHB resistance loci. The data also suggest that resistance to FHB infection and DON accumulation may be controlled, in part, by independent loci and (or) genes. PMID- 12897864 TI - Characterization of the red knot (Calidris canutus) mitochondrial control region. AB - We sequenced the complete mitochondrial control regions of 11 red knots (Calidris canutus). The control region is 1168 bp in length and is flanked by tRNA glutamate (glu) and the gene ND6 at its 5' end and tRNA phenylalanine (phe) and the gene 12S on its 3' end. The sequence possesses conserved sequence blocks F, E, D, C, CSB-1, and the bird similarity box (BSB), as expected for a mitochondrial copy. Flanking tRNA regions show correct secondary structure, and a relative rate test indicated no significant difference between substitution rates in the sequence we obtained versus the known mitochondrial sequence of turnstones (Charadriiformes: Scolopacidae). These characteristics indicate that the sequence is mitochondrial in origin. To confirm this, we sequenced the control region of a single individual using both purified mitochondrial DNA and genomic DNA. The sequences were identical using both methods. The sequence and methods presented in this paper may now serve as a reference for future studies using knot and other avian control regions. Furthermore, the discovery of five variable sites in 11 knots towards the 3' end of the control region, and the variability of this region in contrast to the more conserved central domain in the alignment between knots and other Charadriiformes, highlights the importance of this area as a source of variation for future studies in knots and other birds. PMID- 12897865 TI - Modes of reproduction in Australian populations of Hypericum perforatum L. (St. John's wort) revealed by DNA fingerprinting and cytological methods. AB - Hypericum perforatum L. (St. John's wort) is widely used in homeopathic medicine, but has also become a serious weed in Australia and many other countries. Reproduction in H. perforatum was investigated using markers based on restriction fragment length polymorphism (RFLP) and amplified fragment length polymorphism (AFLP). Between two Australian populations, plants displayed 14 polymorphisms from a total of 22 scorable RFLP markers when genomic DNA was probed with M13 bacteriophage, but individuals within each population exhibited identical RFLP fingerprints. Ninety-four percent of the progeny of four crosses made between the two populations exhibited identical fingerprint and ploidy level to the maternal parent, and probably originated apomictically. Seven seedlings with recombinant RFLP or AFLP fingerprints were found from a total of 121 progeny. Both molecular marker techniques detected the same recombinants from a subset of screened progeny. Cytological analysis showed that the seven recombinants comprised three tetraploids (2n = 4x = 32), three hexaploids (2n = 6x = 48), and one aneuploid (2n - 1 = 31), which suggested that the level of normal reduced embryo sacs was only 2.5%. These results are discussed in relation to the management of invasive populations, and the implications for plant breeding and production of St. John's wort for medicinal purposes. PMID- 12897866 TI - The paternal sex ratio chromosome in the parasitic wasp Trichogramma kaykai condenses the paternal chromosomes into a dense chromatin mass. AB - A recently discovered B chromosome in the parasitoid wasp Trichogramma kaykai was found to be transmitted through males only. Shortly after fertilization, this chromosome eliminates the paternal chromosome set leaving the maternal chromosomes and itself intact. Consequently, the sex ratio in these wasps is changed in favour of males by modifying fertilized diploid eggs into male haploid offspring. In this study, we show that in fertilized eggs at the first mitosis the paternal sex ratio (PSR) chromosome condenses the paternal chromosomes into a so-called paternal chromatin mass (PCM). During this process, the PSR chromosome is morphologically unaffected and is incorporated into the nucleus containing the maternal chromosomes. In the first five mitotic divisions, 67% of the PCMs are associated with one of the nuclei in the embryo. Furthermore, in embryos with an unassociated PCM, all nuclei are at the same mitotic stage, whereas 68% of the PCM-associated nuclei are at a different mitotic phase than the other nuclei in the embryo. Our observations reveal an obvious similarity of the mode of action of the PSR chromosome in T. kaykai with that of the PSR-induced paternal genome loss in the unrelated wasp Nasonia vitripennis. PMID- 12897867 TI - Molecular mapping of Or, a gene inducing beta-carotene accumulation in cauliflower (Brassica oleracea L. var. botrytis). AB - The cauliflower (Brassica oleracea L. var. botrytis) Or gene is a semi-dominant, single-locus mutation that induces the accumulation of high levels of beta carotene in various tissues of the plant, turning them orange. As part of a map based cloning strategy, molecular mapping of the Or gene in the cauliflower genome was undertaken in a mapping population consisting of 195 F2 individuals. By using amplified fragment length polymorphism (AFLP) in conjunction with bulked segregant analysis, we identified 10 AFLP markers closely linked to the Or gene. Four of the most closely linked flanking markers were converted into restriction fragment length polymorphism (RFLP) markers. Mapping of these markers in the mapping population placed two of them at 0.5 cM from the Or locus on one side, while another marker flanked the Or gene at 1.6 cM on the other side. Three of these markers were also successfully converted into sequence-characterized amplified region (SCAR) markers. These PCR-based markers will be useful for a large-scale application in facilitating the positional cloning of the Or gene. PMID- 12897868 TI - Sequence analysis of the ribosomal DNA internal transcribed spacer region in some scallop species (Mollusca: Bivalvia: Pectinidae). AB - The internal transcribed spacer (ITS) region of the ribosomal DNA from the European scallops Aequipecten opercularis, Mimachlamys varia, Hinnites distortus, and Pecten maximus was PCR amplified and sequenced. For each species, three or five clones were examined. The size ranged between 636 and 713 bp (ITS1, 209-276 bp; 5.8S rRNA gene, 157 bp; ITS2, 270-294 bp) and GC content ranged between 47 and 50% (ITS1, 43-49%; 5.8S rRNA gene, 56-57%; ITS2, 44-49%). Variation within repeats was minimal; only clones from M. varia and P. maximus displayed a few variable sites in ITS2. Among scallops, including Chlamys farreri whose ITS sequence appears in databases, significant variation was observed in both ITS1 and ITS2. Phylogenetic analysis using ITS1, ITS2, or both spacer sequences always yielded trees with similar topology. Aequipecten opercularis and P. maximus grouped in one clade and the other three scallops (C. farreri, M. varia, and H. distortus) in another, where M. varia and H. distortus are the more closely related species. These results provide new insights into the evolutionary relationships of scallop species and corroborate the close evolutionary relationship between the tribes Aequipectinini and Pectinini previously deduced from 18S rDNA sequences. PMID- 12897869 TI - Hybridization and introgression of the genomes of Drosophila nasuta and Drosophila albomicans: evolution of new karyotypes. AB - Drosophila nasuta (2n = 8) and Drosophila albomicans (2n = 6) are cross-fertile allopatric sibling chromosomal races of the nasuta subgroup of Drosophila. Hybrids of these races can be maintained for any number of generations. Some of the introgressed hybrid lineages of D. nasuta and D. albomicans, after passing through a transient phase of karyotypic polymorphism, ended up with a stable karyotype whose composition is different from those of the parental races. Such hybrid populations were called cytoraces, in which the chromosomes of D. nasuta and D. albomicans are represented in different combinations. The karyotypic composition of 16 such cytoraces have been presented and discussed with reference to evolutionary strategies such as balancing selection, directional selection, and sex-specific effect on different components of the evolving karyotypes. PMID- 12897870 TI - A combined RFLP-SSR-AFLP map of tetraploid cotton based on a Gossypium hirsutum x Gossypium barbadense backcross population. AB - An interspecific Gossypium hirsutum x Gossypium barbadense backcross population of 75 BC1 plants was evaluated for 1014 markers. The map consists of 888 loci, including 465 AFLPs, 229 SSRs, 192 RFLPs, and 2 morphological markers, ordered in 37 linkage groups that represent most if not all of the 26 chromosomes, altogether spanning 4400 cM. Loci were not evenly distributed over linkage groups, and 18 of the 26 long groups had a single dense region. This paper proposes a partially revised list of the 13 pairs of homoeologous A/D chromosomes of the 2n = 4x = 52 tetraploid cotton genome. The major revisions, which involve the c3-c17, c4-c22, c5-D08, and c10-c20 homoeologous pairs, are based on the mapping of 68 SSR and RFLP loci with a known chromosome assignment, as well as on comparative alignments with previously published G. hirsutum x G. barbadense maps. The overall congruency in the locus orders and distances of common SSR and RFLP loci in these maps allows for an estimation of the consensus length that reaches a minimum of 5500 cM, and is encouraging for future efforts aimed at developing an integrated map of cultivated cotton. The present map also provides a firm framework for precision mapping of Mendelian components of quantitative traits in cotton PMID- 12897871 TI - Molecular genetic linkage maps for allotetraploid Leymus wildryes (Gramineae: Triticeae). AB - Molecular genetic maps were constructed for two full-sib populations, TTC1 and TTC2, derived from two Leymus triticoides x Leymus cinereus hybrids and one common Leymus triticoides tester. Informative DNA markers were detected using 21 EcoRI-MseI and 17 PstI-MseI AFLP primer combinations, 36 anchored SSR or STS primer pairs, and 9 anchored RFLP probes. The 164-sib TTC1 map includes 1069 AFLP markers and 38 anchor loci in 14 linkage groups spanning 2001 cM. The 170-sib TTC2 map contains 1002 AFLP markers and 36 anchor loci in 14 linkage groups spanning 2066 cM. Some 488 homologous AFLP loci and 24 anchor markers detected in both populations showed similar map order. Thus, 1583 AFLP markers and 50 anchor loci were mapped into 14 linkage groups, which evidently correspond to the 14 chromosomes of allotetraploid Leymus (2n = 4x = 28). Synteny of two or more anchor markers from each of the seven homoeologous wheat and barley chromosomes was detected for 12 of the 14 Leymus linkage groups. Moreover, two distinct sets of genome-specific STS markers were identified in these allotetraploid Leymus species. These Leymus genetic maps and populations will provide a useful system to evaluate the inheritance of functionally important traits of two divergent perennial grass species. PMID- 12897872 TI - The genetic diversity of the Vigna angularis complex in Asia. AB - A selected set of accessions of components of the azuki bean (Vigna angularis) complex comprising 123 cultivated accessions and 23 wild or weedy accessions from Bhutan, China (including Taiwan), India, Japan, Korea, and Nepal was analyzed using amplified fragment length polymorphism (AFLP) methodology. Using 12 AFLP primer pairs, 580 unambiguous bands were generated, 313 (53.9%) of which were polymorphic among azuki bean accessions. All 580 bands were used to assess phenotypic (band) and genetic (nucleotide) diversity among the 146 azuki bean accessions. The results indicate five major groups of azuki bean germplasm primarily associated with geographic origin of accessions and their status: wild, weedy, or cultivated. These five groups are (i) Himalayan wild, (ii) Nepal-Bhutan cultivated, (iii) Chinese wild, (iv) Taiwan wild - Bhutan cultivated, and (v) northeast Asian accessions. Within the northeast Asian accessions, three subgroups are present. These consist of (v1) Japanese complex - Korean cultivated, (v2) Japanese cultivated, and (v3) Chinese cultivated accessions. The results suggest domestication of azuki bean occurred at least twice, once in the Himalayan region of southern Asia and once in northeast Asia. The remarkable diversity of azuki bean germplasm in the Himalayan region compared with other regions suggests this is a rich source of germplasm for plant breeding. The results suggest there are important gaps in the germplasm collections of azuki bean and its close relatives from various parts of Asia and that specific collecting missions for Vigna germplasm related to azuki bean in the highlands of subtropical Asia are needed. PMID- 12897873 TI - Soybean seed lustre phenotype and surface protein cosegregate and map to linkage group E. AB - Soybean (Glycine max (L.) Merr.) seeds vary in their surface properties. The lustre, or glossiness, of seeds has been classified into several different phenotypes. Soybean seeds that have a dull lustre or moderate bloom (B) may also have abundant seed surface protein, namely, an abundance of the hydrophobic protein from soybean (HPS). The seed surface protein HPS is an allergen (Gly m 1) that causes asthma in persons allergic to soybean dust. In this study, seed lustre and surface protein content are compared among 71 different soybean cultivars and lines. Dull-seeded phenotypes usually possessed abundant surface protein in comparison to shiny-seeded types, although exceptions were observed. An F2 population of 82 individuals from a cross of OX281 (dull lustre, abundant HPS) and Mukden (shiny lustre, trace amounts of HPS) provided a basis for inheritance studies and genetic mapping analysis. Results indicate that dull seed lustre (B) and surface protein (Hps) loci are dominant Mendelian traits that cosegregate and map to soybean linkage group E. Molecular markers were used to construct a genetic map of 28 cM encompassing B and Hps. Two different molecular markers cosegregated with each of the loci. This study provides additional evidence that Hps may play a role in the adhesion of endocarp tissues to the seed, and offers new methods of selection for seed lustre and surface protein composition in soybean. PMID- 12897874 TI - An integrated physical, genetic and cytogenetic map around the sunn locus of Medicago truncatula. AB - The sunn mutation of Medicago truncatula is a single-gene mutation that confers a novel supernodulation phenotype in response to inoculation with Sinorhizobium meliloti. We took advantage of the publicly available codominant PCR markers, the high-density genetic map, and a linked cytogenetic map to define the physical and genetic region containing sunn. We determined that sunn is located at the bottom of linkage group 4, where a fine-structure genetic map was used to place the locus within a approximately 400-kb contig of bacterial artificial chromosome (BAC) clones. Genetic analyses of the sunn contig, as well as of a second, closely linked BAC contig designated NUM1, indicate that the physical to genetic distance within this chromosome region is in the range of 1000 -1100 kb.cM-1. The ratio of genetic to cytogenetic distance determined across the entire region is 0.3 cM.microm(-1). These estimates are in good agreement with the empirically determined value of approximately 300 kb.microm(-1) measured for the NUM1 contig. The assignment of sunn to a defined physical interval should provide a basis for sequencing and ultimately cloning the responsible gene. PMID- 12897875 TI - Isolation, characterization, and analysis of Leymus-specific DNA sequences. AB - Genomic Southern hybridization using labeled total genomic DNA of Leymus mollis as probe showed intense hybridization signals on all restriction enzyme digested DNA from five species of Leymus Hochst., and four species of Psathyrostachys Nevski. Experiments using the same L. mollis probe, but with unlabeled blocking DNA from Psathyrostachys, showed no hybridization at all. These two genera evidently had the same genomic content. Southern hybridization without blocking allowed identification of DNA fragments abundant in Leymus and Psathyrostachys. Fragments potentially specific to Leymus were cloned. Five repetitive DNA clones from L. mollis and L. arenarius were characterized: pLmIs1, pLmIs44, pLmIs51, pLmIs53, and pLaIs56. These clones hybridized to both Leymus and Psathyrostachys on Southern blots - no clone hybridized to only one of these genera. Both Southern blot and fluorescence in situ hybridization (FISH) experiments showed that all the clones contained dispersed repetitive sequences. They painted all and whole chromosomes uniformly except at centromeres, telomeres, and nucleolar organiser regions. Three of these clones, i.e., pLmIs1, pLmIs44, and pLmIs53, were essentially specific to Leymus and Psathyrostachys - little or no hybridization was detected in other genera such as Triticum, Hordeum, Thinopyrum, or Elymus. Sequence analysis further revealed that the clones were part of retroelements. In particular, the clone pLmIs44 produced hybridization profiles suitable for analysis of genetic relatedness among species. The present study shows that Leymus and Psathyrostachys share the same basic genome, Ns, and therefore provides strong evidence for combining these two genera. PMID- 12897876 TI - The nucleotypic effects of cellular DNA content in cartilaginous and ray-finned fishes. AB - Cytological and organismal characteristics associated with cellular DNA content underpin most adaptionist interpretations of genome size variation. Since fishes are the only group of vertebrate for which relationships between genome size and key cellular parameters are uncertain, the cytological correlates of genome size were examined in this group. The cell and nuclear areas of erythrocytes showed a highly significant positive correlation with each other and with genome size across 22 cartilaginous and 201 ray-finned fishes. Regressions remained significant at all taxonomic levels, as well as among different fish lineages. However, the results revealed that cartilaginous fishes possess higher cytogenomic ratios than ray-finned fishes, as do cold-water fishes relative to their warm-water counterparts. Increases in genome size owing to ploidy shifts were found to influence cell and nucleus size in an immediate and causative manner, an effect that persists in ancient polyploid lineages. These correlations with cytological parameters known to have important influences on organismal phenotypes support an adaptive interpretation for genome size variation in fishes. PMID- 12897877 TI - A single point mutation in Drosophila dihydrofolate reductase confers methotrexate resistance to a transgenic CHO cell line. AB - Sequence analysis of a cDNA encoding dihydrofolate reductase (DHFR) from a selected methotrexate-resistant Drosophila melanogaster cell line (S3MTX) revealed a substitution of Gln for Leu at position 30. Although the S3MTX cells were approximately 1000 fold more resistant to methotrexate (MTX), the karyotype was similar to the parental line and did not show elongated chromosomes. Furthermore, kinetic analysis of the recombinant enzyme showed a decreased affinity for MTX by the mutant DHFR. To determine if the resistance phenotype could be attributed to the mutant allele, Drosophila Dhfr cDNAs isolated from wild type and S3MTX cells were expressed in Chinese hamster ovary (CHO) cells lacking endogenous DHFR. The heterologous insect DHFRs were functional in transgenic clonal cell lines, showing approximately 400-fold greater MTX resistance in the cell line transfected with the mutant Dhfr than the wild type Dhfr. Resistance to other antifolates in the CHO cells was consistent with the drug sensitivities seen in the respective Drosophila cell lines. ELevated Levels of Dhfr transcript and DHFR in transgenic CHO cells bearing the mutant cDNA were not seen. Taken together, these results demonstrate that a single substitution in Drosophila DHFR alone can confer Levels of MTX resistance comparable with that observed after considerable gene amplification in mammalian cells. PMID- 12897878 TI - Rapid genomic changes in interspecific and intergeneric hybrids and allopolyploids of Triticeae. AB - Allopolyploidy is preponderant in plants, which often leads to speciation. Some recent studies indicate that the process of wide hybridization and (or) genome doubling may induce rapid and extensive genetic and epigenetic changes in some plant species and genomic stasis in others. To further study this phenomenon, we analyzed three sets of synthetic allopolyploids in the Triticeae by restriction fragment length polymorphism (RFLP) using a set of expressed sequence tags (ESTs) and retrotransposons as probes. It was found that 40-64.7% of the ESTs detected genomic changes in the three sets of allopolyploids. Changes included disappearance of parental hybridization fragment(s), simultaneous appearance of novel fragment(s) and loss of parental fragment(s), and appearance of novel fragment(s). Some of the changes occurred as early as in the F1 hybrid, whereas others occurred only after allopolyploid formation. Probing with retrotransposons revealed numerous examples of disappearance of sequences. No gross chromosome structural changes or physical elimination of sequences were found. It is suggested that DNA methylation and localized recombination at the DNA level were probably the main causes for the genomic changes. Possible implications of the genomic changes for allopolyploid genome evolution are discussed. PMID- 12897879 TI - A mutation in the spindle checkpoint arresting meiosis II in Brachiaria ruziziensis. AB - Cytological characterization of BRA005568 accession of Brachiaria ruziziensis (2n = 2x = 18) showed a totally unexpected high frequency of abnormal meiotic products, from triads to hexads, and also tetrads with micro nuclei or microcytes. Meiosis I had a low frequency of abnormalities, mainly related to the chiasma terminalization process. In meiosis II, however, frequency of abnormalities increased exceptionally. Early prophase II was normal with the chromosome set enclosed by the nuclear envelope. However, in late prophase II, owing to the breakdown of the nuclear envelope, the chromosomes were scattered in the cytoplasm. Some chromosomes did not reach the metaphase II plate and remained scattered. The behavior of sister cells was inconsistent. While in one cell the chromosomes were totally aligned at the metaphase II plate, in the other they could be found completely scattered, leading to an asynchronous cell division. Cells with scattered chromosomes were unable to progress in meiosis. Thus, anaphase II failed to occur and sister chromatids were not released. Cells with non-aligned chromosomes in the metaphase II plate did not receive the "go ahead" sign to initiate anaphase II. Consequently, the scattered chromosomes produced telophase II nuclei of different sizes in situ. The asynchronous behavior led to the formation of a wide range of meiotic products. Results suggest that the present accession contains a mutation affecting the spindle checkpoint that arrests the second meiotic division. PMID- 12897881 TI - Changes in the pathology reporting of rectal cancer: is it time to adopt synoptic reporting? PMID- 12897882 TI - Pathology reporting of rectal cancer: a national audit. AB - AIM: To audit the quality and completeness of histopathology reports of rectal cancer resections submitted to the National Cancer Registry in 2000. METHODS: All 388 mid- and low-rectal-cancer specimen reports submitted to the Registry were reviewed. Reports were scored according to a pre-defined 'proforma' as to the completeness of the pathological examination and the submitted report. RESULTS: Scores from teaching hospitals, public non-teaching hospitals and private laboratories did not differ significantly. Multiple staging systems were used in 40% of reports and no stage was allocated in 31% of reports. Circumferential margin involvement was recorded in 63% of reports. CONCLUSIONS: No significant differences exist in the quality of pathology reporting of rectal cancer between different laboratory types, either public or private. There is a lack of uniform reporting of rectal cancer stage, with multiple staging systems in use. Circumferential margin involvement is frequently omitted in spite of its documented value as an indicator of quality of rectal cancer surgery, as an important predictor of local recurrence, and its more-recently established value as a marker for distant metastasis and survival. PMID- 12897883 TI - Liver injury in children: causes, patterns and outcomes. AB - AIM: To compare the causes, patterns and outcomes of hepatic trauma in children with those in adults. METHODS: A retrospective audit was conducted of a five-year period from 1996 of adults and children admitted to Christchurch Hospital with liver injuries. Details of age, mechanism of injury, injury severity score (ISS), radiological grade of liver injury, operations and mortality were recorded and analysed. RESULTS: There were 93 liver injuries over the five-year period: 22 in children and 71 in adults. The median age of each group was 7 and 29 years respectively. The most common causes of injury in children were bicycle (7/22) and motor vehicle accidents (MVAs) (7/22). The majority (37/71) of adult injuries were caused by MVAs. The median length of hospital stay was significantly shorter in the paediatric group: 4 days (range 1-12) in children vs 9 days (range 0-52) in adults. CONCLUSIONS: Liver trauma in children has a different spectrum of causes, and results in more severe liver injury than in adults. However, children are more likely to have an isolated liver injury that results in a shorter length of stay in hospital. A nationwide paediatric-injury surveillance system might allow better identification of preventable causes of injury. PMID- 12897884 TI - The Auckland experience with laparoscopic donor nephrectomy. AB - AIMS: To examine the initial experience of laparoscopic donor nephrectomy (LDN) in New Zealand and compare it with open donor nephrectomy (ODN). METHODS: All LDNs performed between June 2000 and June 2002 were reviewed. An equal number of ODNs were reviewed. Data were also collected on the recipients of the grafts. Key clinical data were prospectively collected; remaining data were collected by retrospectively reviewing patient charts. Auckland Hospital databases were accessed for costing analysis. RESULTS: Thirty five cases of each procedure had been performed. There has been 100% LDN graft survival. There was no significant difference in graft function (serum creatinine) at one and 12 months (p = 0.25 and 0.35) between the two groups. There was no significant difference in donor morbidity (26% vs 31%, p = 0.59). LDN resulted in a shorter hospital stay (3 vs 6.5 days, p <0.0001) and convalescence period (3 vs 6 weeks, p <0.0001). LDN was significantly more expensive (13 357 dollars vs 6713 dollars, p <0.0001). CONCLUSIONS: LDN in the New Zealand setting provides effective grafts for renal transplant recipients and is safe for the donor. Advantages for the donor are a shorter hospital stay and convalescence period. The major disadvantage of LDN is its higher cost compared with ODN. PMID- 12897885 TI - Do nonsteroidal anti-inflammatory drugs affect the outcome of patients admitted to hospital with lower gastrointestinal bleeding? AB - AIM: To determine whether the outcome of patients admitted to hospital with lower gastrointestinal bleeding (LGB) is affected by their use of aspirin (ASA) or non aspirin nonsteroidal anti-inflammatory drugs (NANSAIDs). METHODS: A retrospective review of all patients admitted to Wellington Hospital over a four-and-a-half year period from January 1998 with a coded discharge diagnosis that included LGB. Data were collected on requirement for blood transfusion (BT), number of units transfused, drug use, requirement for surgery, and in-hospital mortality. RESULTS: There were 168 admissions to hospital with LGB over the study period of which, after exclusions, 146 formed the basis of this study. The mean age of patients was 69 years, with an equal gender distribution. Fifty three per cent of patients were taking medication known to interfere with platelet function (42% ASA, 18% NANSAIDs, and 7% both). Diverticular disease was the most common diagnosis. Eight patients required surgery for bleeding and there were two in hospital deaths (1.4%). Forty three per cent of admitted patients required BT. Patients taking ASA or NANSAIDs (drug group) were more likely to receive a BT (relative risk 2.7, p <0.00001) than patients in the non-drug group. The median number of units received in transfused patients and requirement for surgery, although higher in the drug group, were not statistically different between the drug the non-drug group. CONCLUSIONS: Patients admitted to hospital with LGB while taking ASA or NANSAIDs are significantly more likely to need BT but use of these agents does not result in a more frequent requirement for surgery. PMID- 12897886 TI - How much has the introduction of laparoscopic surgery changed open surgery? AB - During the last decade of the twentieth century there was a proliferation of laparoscopic surgical procedures. This has been credited with significantly improving the care and outcomes of surgical patients, despite the presence of little evidence to support such claims. Many laparoscopic procedures have developed due to the perception that they are better; however, this perception was not and is not supported by level one data (randomised controlled trial). It is hard to obtain level one data to validate the perception of advantage once a technique has become fully established. At the same time that laparoscopic surgery was developing, many other aspects of peri-operative care changed that influenced outcomes. These changes may account for many of the benefits claimed by laparoscopic surgery. PMID- 12897887 TI - Duodenal somatostatinoma: a rare cause of gastrointestinal bleeding. PMID- 12897888 TI - Non-surgical approach to delayed expansion of traumatic intramural duodenal haematoma. PMID- 12897889 TI - Lignocaine neurotoxicity. PMID- 12897890 TI - Detection of Down's syndrome. PMID- 12897891 TI - Problems with cold and damp housing among Pacific families are not due to ignorance. PMID- 12897892 TI - 6-thioguanine nucleotides and thiopurine methyltransferase activity: important factors determining response to treatment and incidence of adverse effects from azathioprine and 6-MP. PMID- 12897893 TI - Identifying occupational therapists' referral priorities in community health. AB - Occupational therapists in British community mental health teams have been debating how the most effective services can be targeted at the most needy clients. This paper presents the results of a quantitative study that examined 40 British occupational therapists' referral prioritization policies. Results showed half of the participants felt their generic responsibilities, which involved having care co-ordination responsibilities, were too large. Only 25% of participants co-ordinated care for clients whose needs were related to occupational dysfunction. Judgement analysis, that involved regressing the 40 individuals' prioritization decisions onto the 90 respective referral scenarios, was used to statistically model how referral information had been weighted. Group agreement of prioritization was moderate with the reason for referral, history of violence and diagnosis being given the most weighting. Consistency in policy application, as measured by examining prioritization decisions on identical referrals, showed wide variability. Further research is required to identify the optimal and most stable policies within this group. PMID- 12897894 TI - Women's experiences of hassles and uplifts in their everyday patterns of occupations. AB - The aim of this study was to investigate experiences of hassles and uplifts among women. One hundred working mothers were interviewed using the Target Complaints instrument. Content analysis, resulting in both qualitative categories and quantitative variables, was used. Working mothers' hassles were mainly generated by their social, temporal and doing contexts and illustrate the importance of considering women's total patterns of everyday occupations and not focusing one sidedly on the work situation when treating occupation-related ill-health. Women's uplifts were experienced through the social context and by doing such different occupations as going to the movies, cleaning the house, or attending a class. This indicates the appropriateness of using a client-centred approach in interventions with openness to the client's unique situation. Unexpected occupations were identified almost exclusively among the hassles. This is important knowledge for occupational therapists since women will continue to be dual workers and at potential risk of developing unbalanced and detrimental patterns of occupations, in turn causing ill health. PMID- 12897895 TI - A comparison of mothers' and fathers' experience of parenting stress and attributions for parent child interaction outcomes. AB - Parents of children with disabilities are vulnerable to parenting stress, which may place them at physical and psychological risk. However, it is not clear whether fathers experience stress differently to mothers, or whether their experiences are reported less frequently. Additionally, there is little reported on the relationships and gender differences between mothers' and fathers' attributions for parent child interaction outcomes. Parenting stress was assessed in this study using Abidin's (1990) Parenting Stress Index (PSI), and parenting attributions were assessed using the original (Bugental et al., 1989; Bugental and Shennum, 1984), and modified versions of the Parenting Attribution Test, also known as the Child Interaction Survey (CIS) (M-CIS: Esdaile and Greenwood, 1995b). Participants were 53 mothers and 25 fathers of children with disabilities. Having a child with a disability was associated with elevated scores on the PSI; some gender differences were found. Only one significant outcome was found on the assessment of parenting attributions. Thus, the findings suggest that further research is indicated to explore differences in mothers' and fathers' experiences of parenting stress, and the assessment of parenting attributions. The fact that having a child with a disability was associated with elevated scores on the PSI for both mothers and fathers indicates the importance of considering stress management as an integral part of occupational therapy programmes that involve parents of children with special needs. Therapists also need to consider possible gender differences when planning stress management programmes including both mothers and fathers of children with disabilities. PMID- 12897896 TI - Content validity and clinical applicability of the Irena Daily Activity assessment measuring occupational performance in adults with developmental disability. AB - The purpose of this study was to investigate the validity of the Irena Daily Activity (IDA) assessment which measures occupational performance in adults with developmental disabilities. The instrument's content validity was investigated by elucidating its general appropriateness, its clinical applicability and the feasibility of the measurement procedure. Data were collected from a multidisciplinary panel of clinical experts working in day activity centres. Each expert independently judged and reviewed the IDA instrument's content validity. The results show that the content validity index (CVI) values ranged from 0.8 to 1.0 for the instrument's general appropriateness and clinical applicability, and from 0.7 to 1.0 with respect to the significance of the IDA domains' clinical applicability. The applicability of the IDA items were judged as satisfactory (CVI > 80) with respect to their importance for planning treatment and intervention. The feasibility of the measurement procedure was estimated as satisfactory in relation to the items (CVI > 0.70) and satisfactory with respect to the time needed and material used, with CVI values ranging from 0.8 to 1.0. The results of this study support the use of the IDA as a feasible and time efficient assessment that provides insights regarding the occupational performance of adults with developmental disabilities. A limitation of this study was that it could not illuminate all-important aspects of validity. In conclusion, the IDA has the potential to become a reliable and valid clinical assessment, but additional research on psychometric properties is still needed. PMID- 12897897 TI - The voice of detainees in a high security setting on services for people with personality disorder. AB - BACKGROUND: British government Home and Health Departments have been consulting widely about service development for people with ' dangerous severe personality disorder' (DSPD). There has, however, been no consultation with service users, nor is there any user view literature in this area. METHODS: All people detained in one high security hospital under the legal classification of psychopathic disorder were eligible but those on the admission or intensive care wards were not approached. Views of service were elicited using a purpose designed semi structured interview. The principal researcher was independent of all clinical teams. Confidentiality about patients' views was assured. AIMS: To establish views on services from one subgroup of people nominated by the government department as having 'DSPD'. RESULTS: Sixty-one of 89 agreed to interview. With security a given, about half expressed a preference for a high security hospital setting, 20% prison and 25% elsewhere, generally medium secure hospitals. Participants most valued caring, understanding and 'experience' among staff. An ideal service was considered to be one within small, domestic living units, providing group and individual therapies. Some found living with people with mental illness difficult, but some specified not wanting segregated units. Views were affected by gender and comorbidity. CONCLUSIONS: As the sample were all in hospital, the emphasis on treatment may reflect a placement bias. All but five participants, however, had had experience of both health and criminal justice services, so were well placed to talk with authority about preferences. PMID- 12897898 TI - Name change among offender patients: an English high security hospital sample. AB - BACKGROUND: There is scant literature on change of name among psychiatric patients but a more substantial amount on the use of aliases among offenders. No one to our knowledge has explored name changes among offender patients. AIM: This study was undertaken to establish the prevalence of name change among high security hospital patients and to compare those who changed name with those who did not. HYPOTHESIS: It was hypothesized that name change would be associated with male gender, repeated offending and a diagnosis of personality disorder. METHOD: The clinical records of all patients resident in one high security hospital on 31 January 2000 were searched. As all such patients are compulsorily detained, all name changes must be recorded. Any change prior to that date was also noted, together with basic demographic and diagnostic information; supplementary data were obtained from the special hospitals' case register. RESULTS: Seventy-one patients (17%) of the resident population changed names (exclusive of a woman changing her surname on marriage). Name changing was associated with disrupted upbringing. Patients with personality disorder were more likely to change names than those with psychosis, regardless of sex, age or ethnic group. Those with psychosis were more likely to select unusual or symbolic names. CONCLUSIONS: On the evidence of previous literature, people from this serious offender patient population were more likely to change names than other psychiatric patients but less likely than non-mentally-disordered offenders. Changes by people with psychosis seemed related to their illness, whereas changes by those with personality disorder might reflect childhood disruptions in rearing patterns. PMID- 12897899 TI - British forensic psychiatry. PMID- 12897900 TI - The treatment and security needs of patients in special hospitals: views of referring and accepting teams. AB - BACKGROUND: Various studies suggest that 20-30% of patients in England and Wales in high security could be safely managed in medium security but there are no objective criteria and little attention has been paid to differences of opinion. METHOD: The authors compare the views of the referring and receiving team on the security needs of all special hospital patients from two London health authorities. RESULTS: Disagreements were found in only 8% of cases, and they were more likely to occur in patients detained under the legal category of psychopathic disorder. Between 21% and 33% of patients were rated as misplaced by one or other team but these figures include patients who were either already on trial leave in another hospital or on the waiting list for an identified placement. Patients for whom there was agreement on misplacement, with no identified route out of high security, account for 9% of the total. Most patients with treatment-resistant schizophrenia were not receiving atypical anti-psychotic medication. DISCUSSION: It may be more realistic to plan future services on the basis that only 9% of patients are misplaced, rather than the previous estimates that appear to have guided current policy. Patients detained under the legal category of psychopathic disorder present particular problems and there is a need to develop appropriate facilities at medium secure level. In the meantime, no patients should be admitted to high security without consultation with the catchment area service and a jointly agreed plan for future rehabilitation. PMID- 12897901 TI - Evaluation of a group intervention for convicted arsonists with mild and borderline intellectual disabilities. AB - BACKGROUND: The extent to which people with intellectual disabilities (ID) set fires is difficult to ascertain. However, services working with people with ID and offending or quasi-offending histories are increasing the amount of attention that they give to this difficult and perplexing issue. This is due to the real and perceived threat that it presents to society and the seriousness with which it viewed by the criminal justice system. AIM: Against this background there is very little available in the research literature concerning treatment interventions for fire-setting behaviour in this client group, and even less regarding their effectiveness. METHOD: In the current study 14 men and women with mild and borderline ID, convictions for arson and detained in a hospital low secure service were offered and completed a broadly cognitive behavioural, approximately 40-session group-based intervention. The treatment was aimed primarily at reducing fire interest and attitudes associated with fire-setting behaviour. Participants were assessed pre- and post-treatment on a number of fire specific, anger, self-esteem and depression measures. RESULTS: Following treatment, significant improvements were found in all areas assessed, excepting depression. Despite the limitations of the study design, the results provide encouragement and some guidance to practitioners who are required to develop interventions for this challenging, yet much neglected client group. PMID- 12897902 TI - Hepatitis C infections. From diagnosis to treatment. PMID- 12897903 TI - The natural history of hepatitis C virus infection. AB - Chronic hepatitis C virus (HCV) infection is a major health problem worldwide. The natural history of HCV infection is not fully understood. For years, there has been an overestimation of the rate of chronicity in acute HCV. Similar high rates of progression to cirrhosis in chronic HCV were reported. The source of confusion stems from the fact that most acute HCV infections are asymptomatic and never come to medical attention. The consequence of this is that most early studies of natural history reflect the more severe end of the spectrum of the disease. Recent studies reported 43-45% rate of chronicity as opposed to the old rates of 77-85%. Also, the rate of progression to cirrhosis and hepatocellular carcinoma was found to be much lower than previously reported. Multiple factors contribute to the chronicity and progression to cirrhosis, the most important being male gender, age, alcohol intake, and the degree of liver fibrosis on initial biopsy. At least 38% of patients with HCV will manifest symptoms of at least one extrahepatic complication. The most important extrahepatic manifestation is mixed cryoglobulinemia. Other extrahepatic manifestations and their response to antiviral therapy are discussed. PMID- 12897904 TI - The role of the laboratory in the diagnosis and management of chronic hepatitis C patients. AB - Clinical diagnosis of chronic hepatitis C infections is a difficult task. This is due to the insidious nature of the infection and the subclinical and symptomless presentation in the majority of cases. The laboratory plays a principal role not only in the specific diagnosis of the infection but also in assessing the severity and evolution of the liver disease, selection of patients for therapeutic intervention, monitoring treatment and determining the outcome of treatment. To attain these goals, improvements in sensitivity and specificity of various techniques, including molecular diagnostic assays, have been introduced. Most importantly, patients may be excluded if they have conditions that are contra-indicated for treatment as determined by laboratory parameters. In cases of adverse events the drugs may be reduced or withdrawn based on clinical and laboratory results. The improvement over the last decade of laboratory assays has paralleled the success in the sustained response rates reported for hepatitis C virus treatment. A good laboratory provides the tools for diagnosis and treatment essential for good management. This is a multidisciplinary approach involving all branches of pathology. PMID- 12897905 TI - Chronic hepatitis C treatment. Side effects and their management. AB - The current standard of treatment of chronic hepatitis C infection is the combination therapy of pegylated interferon plus ribavirin for 48 weeks for genotype 1 and 4 and 24 weeks for genotype 2 and 3. Side effects such as influenza-like syndrome, gastrointestinal, neuropsychiatric, dermatological and endocrinological symptoms are not uncommon. Laboratory abnormalities such as hematological and biochemical may be frequent. These side effects are compatible with treatment continuation if symptoms are managed carefully. The adverse effects are dose dependent and often reversible. Premature withdrawal rates can be reduced if side effects are identified early. It is, however, pertinent to target treatment to early responders and avoid side effects in patients who have low predictive response to treatment. PMID- 12897906 TI - Hepatis C virus genotype 4. AB - Hepatitis C virus genotype 4 chronic hepatitis predominates in the Eastern Mediterranean region of the world. In the Kingdom of Saudi Arabia, as high as 62% of all cases are due to genotype 4. Results of efficacy and safety clinical trials using pegylated-interferon plus ribavirin for chronic hepatitis C patients have shown great promise, but treatment of early responders should be maintained at 180 mg of pegylated-interferon alfa-2a plus 1000-1200 mg of ribavirin for 48 weeks unlike the 24 weeks recommended for genotypes 2 and 3. Hepatitis C genotype 4 may not be as "difficult to treat" as initially thought. PMID- 12897907 TI - Hepatitis C virus seroprevalence rate among Saudis. AB - OBJECTIVE: The aim of the study is to determine the seroprevalence of hepatitis C virus (HCV) in blood donors, children, pregnant women, hemodialysis patients and drug addicts in Saudi Arabia. METHODS: Using third generation enzyme immunoassay kits, we have screened Saudi cohorts of all ages and sexes, namely infants, pre school, school children, young adults and adults (blood donors and antenatals) for antibodies to HCV. We have also reviewed HCV seroprevalence data among high risk groups from 1998 to 2002. RESULTS: An overall 1.1% (6313 out of 557813) seroprevalence rate was determined among Saudi blood donors; 0.1% (5 out of 3854) in Saudi children; and 0.7% (22 out of 3127) among pregnant women. Hemodialysis patients remain at highest risk of infection at 55.7% whereas intravenous drug addicts have 14% exposure rate. CONCLUSION: We conclude that the present public health schemes have been effective in reducing hepatitis C infection in the general community in the Kingdom of Saudi Arabia but the infection among high risk groups remain a major problem that needs to be actively addressed. PMID- 12897908 TI - Hepatitis C genotypes/subtypes among chronic hepatitis patients in Saudi Arabia. AB - OBJECTIVE: The object of this study is to determine the molecular epidemiology of hepatitis C virus (HCV) in the Kingdom of Saudi Arabia (KSA). METHODS: Four hundred and ninety-two histological proven chronic HCV patients prospectively recruited from all regions of KSA, between November 1999 and March 2002, were genotyped and subtyped using amplified products of specific primers from the 5 UTR region in a reverse transcription polymerase chain reaction (Roche Diagnostics, Switzerland) followed by a reverse hybridization technique (Innolipa HCV II [Innogenetics, Belgium]). RESULTS: Sixty-two percent of Saudis were found to be genotype 4. Other genotypes were 1 (24.1%); 2 (7.4%); 3 (5.9%); 5 (0.3%); and 10 (0.3%). There were no differences in distribution patterns between sexes and ages. All regions showed similar distribution except the Eastern region where subtype 2a/c seem to have emerged. Diabetic patients and those with a history of blood transfusion had the same pattern as those with community acquired HCV. Among the non-Saudis (mostly Egyptians), genotype 4 predominated (88%). CONCLUSION: We conclude that 86% of Saudi chronic hepatitis C cases are due to genotypes 1 and 4. Since these are considered "difficult to treat" an aggressive approach to management using combination therapy of pegylated interferon plus ribavirin for 48 weeks should be considered for all cases of chronic hepatitis C until genotyping proves otherwise. PMID- 12897961 TI - Alpha-neurotoxins of Naja atra and Naja kaouthia snakes in different regions. AB - Recent studies have shown that there are geographic variation of alpha neurotoxins in Naja kaouthia, but the cause is not clear yet. In this work, venoms were collected from adult Naja atra in Zhejiang Province and Naja kaouthia in Yunnan Province, well identified by morphological characters and cytochrome b gene analysis in summer season to avoid age and seasonal variation in the venom composition. Then alpha-neurotoxins were purified and cloned from these two kinds of snakes. Three alpha-neurotoxins from Naja kaouthia (Yunnan) and two from Naja atra (Zhejiang) were identified. Together with previously reported alpha neurotoxins in Naja kaouthia (Thailand) and Naja atra (Taiwan Province), it was found that the alpha-neurotoxins of Naja kaouthia in Yunnan Province were similar to those of Naja atra in Zhejiang and Taiwan Provinces, but different from those of Naja kaouthia in Thailand. This result can hardly be explained by population phylogeny or geographic distance. It might be due to the different climate, habitat and prey in Thailand in comparison with those in Yunnan, Zhejiang and Taiwan Provinces. PMID- 12897962 TI - Jerdonase, a novel serine protease with kinin-releasing and fibrinogenolytic activity from Trimeresurus jerdonii venom. AB - A novel kinin-releasing and fibrin(ogen)olytic enzyme termed jerdonase was purified to homogeneity from the venom of Trimeresurus jerdonii by DEAE Sephadex A-50 anion exchange, Sephadex G-100 (superfine) gel filtration and reverse-phase high performance liquid chromatography (RP-HPLC). Jerdonase migrated as a single band with an approximate molecular weight of 55 kD under the reduced conditions and 53 kD under the non-reduced conditions. The enzyme was a glycoprotein containing 35.8% neutral carbohydrate. The N-terminal amino acid sequence of jerdonase was determined to be IIGGDECNINEHPFLVALYDA, which showed high sequence identity to other snake venom serine proteases. Jerdonase catalyzed the hydrolysis of BAEE, S-2238 and S-2302, which was inhibited by phenylmethylsulfonyl fluoride (PMSF), but not affected by ethylenediaminetetraacetic acid (EDTA). Jerdonase preferentially cleaved the A alpha-chain of human fibrinogen with lower activity towards B beta-chain. Moreover, the enzyme hydrolyzed bovine low-molecular-mass kininogen and releasing bradykinin. In conclusion, all results indicated that jerdonase was a multifunctional venom serine protease. PMID- 12897963 TI - Antigen-expressed recombinant Salmonella typhimurium driven by an in vivo activated promoter is capable of inducing cellular immune response in transgenic mice. AB - To explore the approaches and mechanisms for reversing the immune tolerance in transgenic mouse, and the pathogenicity of hepatitis G virus (HGV), the promoter of phoP-activated gene (P(pagC)) of Salmonella typhimurium was used as a transcriptionally regulating element to construct an attenuated S. typhimurium expressing HGV NS3. The recombinant S. typhimurium was orally administered to HGV transgenic mice. As the results, HGV antigen in serum and liver as well as HGV mRNA in liver were decreased significantly, although the serum anti-HGV NS3 remained undetectable as the control transgenic mice. The spleen cell proliferation, in vitro HGV NS3 specific CTL, and IFN-gamma assays with the primed cultured splenocytes indicated the induction of Th1 immune responses in those administered transgenic mice. Adoptive transfer of fractionated primed spleen cells to the transgenic mice showed that T lymphocytes were responsible for, maybe through IFN-gamma, the down-regulation of HGV mRNA transcription. Histological examination found no significant inflammatory changes in liver of the transgenic mice. These findings suggested that the oral inoculation of the HGV NS3-expressed attenuated S. typhimurium driven by an in vivo-activated promoter should be a simple and effective approach for potential treatment of chronic viral infection. PMID- 12897964 TI - [Assessment of the Escherichia coli Tat protein translocation system with fluorescent proteins]. AB - The possibility of using fluorescent proteins as probes to study the twin arginine translocation (Tat) system was assessed in Escherichia coli. When fused to the twin-arginine signal peptide of trimethylamine N-oxide reductase, the DsRed2 red fluorescent protein from the Discosoma sp. was successfully synthesized and folded in E. coli cells. However, RR-DsRed2 aggregated inside the cells. Therefore, although DsRed2 has been engineered from DsRed for faster maturation and lower non-specific aggregation, it is still not compatible with Tat-dependent translocation. In contrast, the jellyfish green fluorescent protein (GFP) was efficiently exported into periplasm even when the RR motif was changed to KR or RK. These results show that GFP can be used as an efficient reporter protein to study Tat system, but DsRed2 is not suitable for such purpose because of its aggregation property. In addition, when the protein concentration was similar, the fluorescence intensity of KR-GFP and RK-GFP decreased compared with RR-GFP, which would suggest that the twin-arginine signal peptide is not only essential for mediating protein translocation, but also important for the folding of down-stream protein. PMID- 12897965 TI - [Estimating coarse gene networks from yeast gene expression time series]. AB - Gene networks is the collection of gene-gene regulatory relations at the expression level. In this study, a combined approach of the linear transcriptional modeling, identification of promoter elements and gene co expression clustering is developed to decipher yeast gene networks from expression time series. The cell must reorganize the genomic expression to programs required for growth and survival in each environment. The expression of many genes is regulated by environmental stress. The products of many genes that induced in the environmental stress are involved in metabolism of carbohydrates, structural repairs and even sporulation. Interestingly, it is identified that transcription factors Mcm1 and Dal82 matched their binding sites TT[bond]CC[triple bond]T[double bond]GGAAA and TGAAAAWTTT in cell cycle progression and environmental stress response, respectively. These conclusions agree with the known observations. The results indicate that the approach may be useful for modeling gene networks from microarray data. PMID- 12897966 TI - [Overexpression of STAT3 in COS7 cells causes prominent morphological changes]. AB - Signal transducer and activator of transcription 3 (STAT3) plays a central role in mediating signals related to cell growth, differentiation, survival and movement. In this report, the results of immunocytochemistry and Western blot analysis showed that the active form of Stat3 protein existed in COS7 cells. The endogenous Stat3 protein could also induce the expression of m67-sequence directed reporter genes. The transient transfection of Stat3 cDNA into COS7 cells increased Stat3 protein level and the expression of the reporter gene. The overexpression of Stat3 protein in COS7 cells caused prominent cell morphological changes. These cells had a much larger cell body, extended long processes with branches, lamellipodia and filopodia. These results suggest that Stat3 protein may play important roles in cell adhesion, migration and cytoskeleton reorganization. PMID- 12897967 TI - [Separation of hydrophobic NAD(P)H dehydrogenase subcomplexes from cyanobacterium Synechocystis PCC6803]. AB - Many efforts have been paid to the separation of an integrated NA(D)PH dehydrogenase (NDH) complex. Several hydrophilic subcomplexes of NDH have been purified from the cyanobacterium Synechocystis PCC6803. However, no hydrophobic NDH subcomplex has ever been separated from cyanobacteria yet. In this paper, two NDH subcomplexes were separated from n-dodecyl beta-D-maltoside(DM)-treated whole cell extracts of Synechocystis PCC6803 by anion exchange chromatography and gel filtration. Both subcomplexes contained the hydrophobic subunit NdhA, suggesting that they were hydrophobic NDH subcomplexes. Of the two subcomplexes, only one subcomplex contained NdhH. These subcomplexes showed NADPH-nitroblue tetrazolium (NBT) oxidoreductase activity and could specifically oxidize NADPH when several quinone analogues were used as electron acceptors, such as ferricyanide, 2,5 dibromo-3-methyl-6-isopropyl-p-benzoquinone (DBMIB), 2,6-dichlorophenol indophenol (DCPIP), duroquinone, ubiquinone-0 (UQ-0), etc. PMID- 12897968 TI - [Knockout of tyrR gene in Escherichia coli and its effects on the phenylalanine biosynthesis]. AB - TyrR is a global regulation protein encoded by tyrR gene which controls several transcriptional units involving biosynthesis and transportation of aromatic amino acids in Escherichia coli. In this work, the tyrR gene was knocked out by a double-cross homologous recombination. The tyrR- mutant was verified by structural identification by PCR and sequencing, and functional demonstration using lacZ reporter gene. In tyrR- mutant, the activities of two key enzymes in the phenylalanine biosynthesis pathway, whose expression was controlled by TyrR, DAHPS and AT, had been shown to elevate by a 1.08-fold and 2.70-fold compared with the parent strain, respectively. The yield of phenylalanine biosynthesis in the mutant was 1.59 times higher than that of wild type strain. The repression on the transcription of aroP, encoding an aromatic amino acid permease, was eliminated, resulting in a 70.2% increase of the aromatic amino acid transportation in tyrR- mutant strain. PMID- 12897969 TI - [A novel approach for peptide identification by tandem mass spectrometry]. AB - High throughput scoring algorithms that are used to find the match of a tandem mass spectrum to a predicted mass spectrum of a peptide within a database have been applied in shotgun proteomics. However, these algorithms could produce a significant number of incorrect peptide identifications. Here a novel approach was developed to scoring tandem mass spectra against a peptide database, in which fragment ion probabilities, number of enzymatic termini of candidate peptides, matching quality and match pattern between experimental and theoretical spectrum were considered. Benchmarking the novel scorer on a large set of experimental MS/MS spectra, it is demonstrated that PepSearch performs significantly better than the widely used software SEQUEST. The PepSearch software is available at http://compbio.sibsnet.org/projects/pepsearch. PMID- 12897970 TI - Analysis of rat electroencephalogram during slow wave sleep and transition sleep using wavelet transform. AB - The dynamic features of rat EEGs collected during slow wave sleep (SWS) and transition sleep (TS) were investigated in both time and frequency domains using wavelet transform based on multi-resolution signal decomposition. EEGs of freely moving rats were recorded with implanted electrodes and then decomposed into four components of delta, theta, alpha and beta using wavelet transform. The power and power percentage of each component were calculated as functions of time. In SWS EEGs, the results showed that there existed as much as 26.2% +/- 7.7% time duration in which the delta power percentage was less than 50%. In addition, the powers of other three components in small delta EEGs were significantly larger than those in large delta EEGs. This result revealed a reciprocal relationship between delta oscillation and spindle oscillation. Comparatively, the conventional method of FFT based power spectrum could only show a delta power dominating (70.6% +/- 6.4%) spectrum of SWS EEGs. In the non-stationary TS EEG, spindle and non-spindle segments were distinguished based on the wavelet components of theta and alpha, and then the average duration of the spindles was estimated. In conclusion, the wavelet transform may be useful in developing novel quantitative time-frequency measures of sleep EEGs as valuable complements of conventional FFT method to analyze the transient changes in sleep EEGs induced by physiological, pathological or pharmacological conditions. PMID- 12897971 TI - Interaction of C17orf25 with ADP-ribose pyrophosphatase NUDT9 detected via yeast two-hybrid method. AB - The gene C17orf25 was isolated from the liver by RACE PCR. nudt9 gene was screened by yeast two-hybrid method in MatchMaker human HeLa cDNA library. NUDT9 is an enzyme that has pyrophosphatase activity with ADP-ribose as its substrate. Fusion expression of C17orf25 and GFP and computer analysis showed that C17orf25 was probably located in mitochondria. Furthermore, C17orf25 may suppress the cell growth by interaction with NUDT9. PMID- 12897972 TI - [Morphological observation on fibronectin fibrils surrounding human breast carcinoma cells by atomic force microscopy]. AB - This report aimed at investigation on the application of atomic force microscope (AFM) in research on interaction between cells and extracellular matrix, and function of extracellular matrix. Distribution and array of fibronectin fibrils surrounding human breast carcinoma MCF-7-R cells were observed by atomic force microscope, and comparison of AFM with other traditional techniques was performed. Whole and local AFM morphological images of several human breast carcinoma MCF-7-R cells and fibronectin fibrils around cells were obtained by AFM. Distribution and array of fibronectin fibrils were found to be very regular, and the regulated array coordinated with functions of fibronectin fibrils very well. Owing to advantages of simple sample preparation and very high resolution, AFM is an ideal tool to in situ observation of extracellular matrix. PMID- 12897973 TI - [Production and application of rabbit anti-imitative spider dragline silk protein polyclonal antibody]. AB - A synthetic spider dragline gene s600 was cloned into fusion protein expression vector pGEX-KG and expressed in Escherichia coli. Protein S600 was purified and rabbit antiserum was prepared. Amino acid composition analysis confirmed the right expression of S600. Western blot analysis revealed that anti-S600 antiserum could react with natural spider silk, so the synthetic dragline protein, designed by the authors, shares similar immunological characteristics with the natural spider silk. An ELISA system was also established for the quantitative detection of synthetic dragline protein expression in silk gland (or in the cocoon) of transgenic silkworm. PMID- 12897974 TI - [Expression and characterization of Kringle 1-5 domains of human plasminogen]. AB - The cDNA encoding Kringle 1-5 domains of human plasminogen (designated as K1-5), obtained from HepG2 by RT-PCR, was cloned into expression vector pHIL-S1. The recombinant plasmid pHIL-K1-5 was transformed into Pichia pastoris GS115 and the recombinant yeast was induced by methanol to express the recombinant protein. The expressed protein was purified by lysine affinity chromatography. The recombinant K1-5 inhibited the growth of bovine aortic endothelial cells (BAEC) stimulated by the basic fibroblast growth factor (bFGF), in a dosage-dependent manner, with a half maximal concentration of 14 mg/L. And rhK1-5 inhibited 47% of the BAEC migration stimulated by bFGF at the concentration of 50 mg/L. rhK1-5 also affected the cell cycle of BAEC and caused G(0)-G(1) arrest at the concentration of 14 mg/L. PMID- 12897975 TI - [Characterization of the copy number of RIRE10 retrotransposon and transcriptional activity of its LTR in rice genome]. AB - A full-length Ty3-like retrotransposon, named RIRE10, was identified on the long arm of chromosome 4 in rice genome. The internal region between two LTRs had another open reading frame in the region upstream of gag-pol sequence. The transcripts from LTR region were detected by Northern blot hybridization and RT PCR. To assess the activity of RIRE10 in rice genome, the copy number of its internal region and long terminal repeat (LTR) domain were determined by dot blot analyses. Nearly 900 solo-LTR of the RIRE10 retrotransposon exist in rice genome, apart from those LTRs that flank 65 intact RIRE retrotransposons. Based on the experimental results, the retrotransposition of RIRE10 was speculated to be influenced by two factors: transcriptional activity of LTR region and homologous recombination resulting in solo-LTR. PMID- 12897976 TI - [Expression and purification of recombinant SARS coronavirus spike protein]. AB - A novel coronavirus (SARS-coronavirus, SARS-CoV) was discovered in association with cases of severe acute respiratory syndrome (SARS) recently. The first step in coronavirus infection is binding of the viral spike protein to certain receptor on host cells. The spike protein is the main surface antigen of the coronavirus and there should be antibodies against spike protein in patients serum. Thus, to develop and expression protein fragment from spike protein gene are the purposes of this experiment. Partial spike gene fragments (751-1925 bp, 2005-3410 bp, 1-1925 bp and 32-3659 bp) and its intact gene were cloned into pET32 or pGEX vectors, and transformed into competent Escherichia coli BL21(DE3) (pLysS), respectively. 63, 78, 98, 160 and 164 kD fusion proteins were successfully expressed with amounts of 35%, 34%, 24%, 17% and 5% of total cell protein. The soluble parts of the cell crude extract were then partially purified by GST affinity chromatography. PMID- 12897977 TI - Can radial bone mineral density and quantitative ultrasound measurements reduce the number of women who need axial density skeletal assessment? AB - The purpose of this study was to investigate the clinical usefulness of forearm bone mineral density (BMD) and speed of sound (SOS) at the phalanx and radius as pre-selection tests to identify women with low BMD at the axial skeleton. BMD was measured by dual-energy X-ray absorptiometry (DXA) in the forearm, lumbar spine and femoral neck. SOS at the radius and phalanx was measured using a multisite quantitative ultrasound (QUS) device. Measurements were performed on 524 consecutive women referred for the assessment of BMD. Women with a T-score <-1 and T-score < or =-2.5 at either spine or femoral neck were identified, and T score cut-off values for the forearm DXA and QUS variables were determined. Cut off values for the forearm BMD estimated to detect normal women and those with T score <-1 at the axial skeleton identified a total of 82% of subjects with 91% certainty. Cut-off values for the forearm BMD determined to detect women with T score >-2.5 and those with osteoporosis allowed the identification of 62% of the study population with 90% certainty. Cut-off values for the phalangeal and radial SOS estimated to detect normal women and those with T-score <-1 at the axial skeleton identified a total of 49% and 1% of subjects, respectively. Cut-off values estimated for QUS variables to detect women with T-score >-2.5 and those with osteoporosis at the axial skeleton either failed to detect subjects with sufficient certainty (phalangeal SOS) or detected a negligible percentage of patients (radial SOS). In conclusion, forearm BMD may be used as a pre-selection test to identify women with low BMD at the axial skeleton, thus enabling reduction of the number of women who need axial BMD assessment. SOS of the phalanges and radius appears to have less value in the detection of the women with low axial BMD. PMID- 12897978 TI - Comparison of a simple clinical risk index and quantitative bone ultrasound for identifying women at increased risk of osteoporosis. AB - Osteoporosis is a growing problem in Asia, and early identification of at risk subjects for preventive measures is likely the most cost-effective method for managing this disease in developing countries. Patients with low bone mineral density (BMD) have a high risk of future fracture. However, access to BMD measurements is limited in many areas of Asia, and inexpensive methods of targeting high-risk patients for BMD measurements would be valuable. We compared two methods, a simple clinical risk assessment tool, the Osteoporosis Self assessment Tool for Asians (OSTA), and quantitative bone ultrasound (QUS) in identifying subjects with low BMD by DXA in 722 southern Chinese postmenopausal women recruited from the community in Hong Kong. Using the published cutoff value of -1 (versus 0 or higher) for OSTA to identify subjects with femoral neck BMD T score < or =-2.5, basing on our local population peak young mean value, the sensitivity and specificity was 88% and 54% respectively. The optimal cutoff T score of -2.35 for QUS yielded sensitivity and specificity values of 81% and 65%, respectively. The AUC for QUS was 0.78, which was not significantly different from that of 0.80 for OSTA. Both OSTA and QUS correlated significantly with BMD at the femoral neck (0.62 and 0.36, respectively, P both <0.001). When these cut off values were used to identify subjects with either lumbar spine or femoral neck BMD T-score < or =-2.5, the sensitivity and specificity was 79% and 60%, respectively, for OSTA, and 69% and 70%, respectively, for QUS. Combining QUS with OSTA improved the sensitivity to 91%, but the specificity was reduced to 44%. We conclude that the simple clinical risk assessment tool OSTA is a free and effective method for identifying subjects at increased risk of osteoporosis, and its use could facilitate the appropriate and more cost-effective use of bone densitometry in developing countries. PMID- 12897979 TI - Identification of risk factors for future fracture in patients following distal forearm fracture. AB - The challenge for healthcare systems around the world is delivering timely preventative strategies to subjects most likely to develop fragility fractures. The success or failure of national campaigns will be determined at local level, and many studies to date have found under-utilization of osteoporosis treatment strategies due to reduced public and healthcare professional awareness. An important link between the at-risk patients and their appropriate therapy is their identification and assessment. In the context of a medium sized secondary care provider, this study examined the efficiency of a nurse-led service for assessing women following distal forearm fracture, looking for additional risk factors for future fracture and also looking at uptake of the assessment programme among the target population. Two hundred and two women, median (range) age 69 (50-91) years presented with distal forearm fracture over a 9-month period. Fifty-four subjects did not participate in the study: they were significantly older than those who did ( P<0.002). One hundred and forty-eight subjects underwent assessment of falls risk, future hip fracture risk and calcaneal ultrasound attenuation measurement: 107 attended for assessment at the hospital and 41 required a health professional to visit at their place of residence-the latter group were significantly older ( P<0.0002). Of 148 subjects assessed, 76 (51%) had one or more risk factors for a fall: these subjects were significantly older ( P<0.0002) and more likely to have required a home visit ( P<0.002). Twenty-seven (18%) were considered at risk of a future hip fracture: again they were older and more likely to have required a home visit ( P<0.0005). Ninety-one subjects (61%) had a calcaneal ultrasound attenuation T-score below 2.0, and were referred for bone densitometry: again, they were older ( P<0.0002) and more likely to have required a home visit ( P<0.05). Bone densitometry demonstrated osteoporosis in 47 (52%) of those with low attenuation, osteopenia in 31 and normal bone density in 13. Of 76 subjects deemed to be at risk of falls, 31 (41%) had osteoporosis demonstrated on bone densitometry and of those at risk of future hip fracture, 12 (44%) had osteoporosis. Only nine (22%) subjects who received home visits had no risk factors for falls or hip fracture and normal bone density, compared to 45 (42%) of those who attended hospital. This study has demonstrated that the efficiency of a program to assess additional risk of future fracture in a population who have already fractured may be influenced by where the assessment is delivered: it may be that the patients at greatest risk did not avail of the service. PMID- 12897980 TI - Bone mineral density and bone metabolism in Duchenne muscular dystrophy. AB - Very few studies on bone mineral density and bone metabolism in Duchenne muscular dystrophy (DMD) have been reported. DMD is a severe, progressive muscular disease resulting in death at a young age. No specific therapies are available, but corticosteroids induce improvement and slower progression of the disease. However, long-term steroid therapy is a serious risk factor for osteoporosis. This study was aimed at evaluating bone mineral density and calciotropic hormones in a group of children affected by DMD, with or without steroid therapy. Bone mineral density was measured by DXA scan on lumbar spine and total body. Evaluation of calcium, phosphorus, bone turnover markers and calciotropic hormones was performed. Thirty-two children affected by DMD were studied: twenty two on long-term prednisone therapy, ten not taking corticosteroids. Bone mineral density was lower than normal for age in all patients, and even lower in the group of steroid-treated children. Trunk and lower limb bone mineral densities were more reduced than upper limb mineral density, especially in the steroid treated subjects. A marked reduction in spine bone mineral density, hypocalciuria, low 25-hydroxyvitamin D levels, and increased bone turnover markers were observed, and even these especially in the steroid-treated group. In conclusion, decreased bone mineral density and derangement of calcium metabolism were present in DMD patients, and were worsening during corticosteroid therapy. It is thus recommended that bone and mineral metabolism be carefully evaluated in patients with DMD, so that appropriate measures could be taken, especially now that chronic corticosteroid therapy is frequently given. PMID- 12897981 TI - Can a post-operative brace in slight hyperextension prevent extension deficit after anterior cruciate ligament reconstruction? A prospective randomised study. AB - It has been our observation that post-operative anterior cruciate ligament (ACL) braces together with the post-operative bandages do not always allow the knee to reach full extension. In ten uninjured knees with known hyperextension, the knees were bandaged in the same way as after an ACL-reconstruction. The knees were then studied radiologically in a Hypex brace set at 0 degrees, -5 degrees and -10 degrees of knee extension. Not a single knee was found to be straight in the brace set at 0 degrees. At -5 degrees most of the knees were straight or in slight hyperextension. It took -10 degrees to get all knees straight or in hyperextension. In a prospective randomised study 44 patients who underwent an arthroscopic ACL-reconstruction with a bone patellar tendon bone graft were randomised to use either a brace set at -5 degrees or a straight brace (0 degrees ) for at least the first three postoperative weeks. Before and three months after surgery range of motion was determined, using a goniometer with long arms, and sagittal knee laxity was measured with a KT-2000 arthrometer at manual max. Pre- and post-operative pain was evaluated with the Visual Analogue Scale (VAS). The same examiner (blindfolded to what type of brace was used) performed all the measurements. At three months, two of the 22 patients with the brace set at -5 degrees and twelve of the 22 patients with the straight brace had a loss of full extension of 2 degrees or more ( p<0.001). No significant differences were found between the groups in terms of knee flexion, sagittal knee laxity or post operative pain. Although extension deficit after ACL-reconstruction can be prevented also in other ways, a Hypex brace set at -5 degrees seems to be an easy way of ensuring full knee extension. PMID- 12897982 TI - Initial fixation strength of interference nail fixation for anterior cruciate ligament reconstruction with patellar tendon graft (experimental study). AB - The objective of this study was to evaluate initial fixation strength of a new interference nail fixation in anterior cruciate ligament reconstruction using bone-patellar tendon-bone graft. Human cadaver knees were used. Fixation strengths ranged from 500 N to 600 N (mean 550 N). This corresponds to loads in the graft during aggressive rehabilitation. No slippage occurred at fixation site of specimens. Most of the failures appeared from the femoral side with tendon ruptures. With respect to primary fixation strength, interference nail fixation is a reasonable alternative for anterior cruciate ligament reconstruction with bone-patellar tendon-bone graft. PMID- 12897983 TI - Observed kneeling ability after total, unicompartmental and patellofemoral knee arthroplasty: perception versus reality. AB - Kneeling is an important function of the knee, but little information is available on ability to kneel after different knee arthroplasty procedures. Previous work has asked patients about their kneeling ability; in this study it was objectively assessed. One hundred and twenty two patients - 38 having had total knee replacement (TKR), 53 unicompartmental knee replacement (UKR), 31 patello-femoral replacement (PFR) - were observed trying to kneel at 90 degrees on a chair, at 90 degrees on the floor, and at 120 degrees on the floor. Only 37% of patients thought they could kneel, whereas 81% were actually able to kneel ( p<0.001). Ability to kneel on the chair and on the floor at 90 degrees was significantly better than perceived ability for all prosthesis types ( p<0.001). Kneeling at 120 degrees showed no difference between perception and reality except for the PFR group ( p<0.05). In all positions, increased range of movement significantly improved kneeling ability ( p<0.001). Kneeling ability in men was significantly better than in women ( p<0.001). Patient-centred questionnaires do not accurately document kneeling ability after knee arthroplasty. PMID- 12897984 TI - A preliminary report on prevalence of acetabular labrum tears in sports patients with groin pain. AB - To the best of our knowledge, this prospective study is the first to investigate the prevalence of acetabular labrum tears in athletes presenting with groin pain. Eighteen athletes who presented to our sports clinic with groin pain, underwent clinical assessment and magnetic resonance arthrography (MRa) to detect presence or absence of acetabular labrum tears. Ethical committee approval and informed consent was obtained from each patient. In four out of these eighteen athletes (22%) the MRa demonstrated the presence of acetabular labrum tear. Three of them underwent arthroscopic debridement of their acetabular labrum tears and returned to their sporting activities within 8 months. Clicking sensation of the hip was a sensitive (100%) and specific (85%) clinical symptom to predict labral tears. The internal rotation-flexion-axial compression manoeuvre was sensitive (75%) but not specific (43%). The Thomas test was neither sensitive nor specific. The conclusion of the study is that acetabular labrum tears can be a common cause of groin pain in athletes. Sports clinicians managing athletes with groin pain have to be well aware of the condition. PMID- 12897985 TI - Mechanical properties of the rotator cuff: response to cyclic loading at varying abduction angles. AB - The rotator cuff is loaded under static as well as dynamic conditions. Whilst the static properties of the rotator cuff muscle-tendon junctions have been reported, the dynamic mechanical behaviour has not. This study reports the dynamic mechanical properties with varying abduction angles in a human cadaver rotator cuff. No significant effect was found with varying the angle of testing or in the presence of a tear in the tendon. The supraspinatus was found to be the stiffest of the rotator cuff tendons followed by the subscapularis and infraspinatus. PMID- 12897986 TI - Calcium pyrophosphate dihydrate crystal deposition disease mimicking malignant soft tissue tumor. AB - Calcium pyrophosphate dihydrate (CPPD) crystal deposition disease has multiple clinical features with variable courses creating several pitfalls in clinical diagnosis. There are number of reported cases mimicking malignant skeletal tumors such as chondrosarcoma. However, no case of CPPD disease with radiographic noncalcified soft tissue mass has been reported in the literature. Here we report a case of CPPD disease clinically mimicking soft tissue tumor with its magnetic resonance imaging appearance and histopathology. PMID- 12897987 TI - TGF-beta1 impairs homocysteine metabolism in human renal cells: possible implications for transplantation. AB - We hypothesized that TGF-beta1 influences the metabolism of homocysteine (Hcy) and increases its cellular export, which may lead to hyperhomocysteinemia in patients with renal transplants. We exposed human renal proximal tubule epithelial cells (huRPTECs) to different concentrations of TGF-beta1, IL-1alpha, IL-10, or methionine and measured total Hcy (tHcy) in culture supernatants. We then examined the relationship between plasma levels of tHcy and TGF-beta1 in renal graft recipients. In multivariate analysis, the factors mediator (TGF beta1, IL-1alpha, IL-10), mediator concentration, methionine concentration, and "mediator x concentration" interaction independently influenced tHcy concentrations in culture supernatants. A 31% increase in tHcy was observed after exposure of huRPTECs to TGF-beta1 compared to medium alone. However, TGF-beta1 plasma levels in kidney graft recipients showed no independent association with tHcy plasma concentrations. We demonstrated that the release of Hcy from huRPTECs is enhanced by TGF-beta1, but that TGF-beta1 plasma levels in renal graft recipients show no independent relationship with hyperhomocysteinemia. PMID- 12897988 TI - Weight loss of respiratory muscles during mechanical ventilation. PMID- 12897989 TI - Adult presentation of MCAD deficiency revealed by coma and severe arrythmias. AB - We report the case of a 33-year-old man who presented with headaches and vomiting. Soon after admission he became drowsy and agitated, developed ventricular tachycardia and his neurological state worsened (Glasgow coma score 6). Blood analysis showed respiratory alkalosis, hyperlactacidemia (8 mmol/l), hyperammonemia (390 micro mol/l) and hypoglycaemia (2.4 mmol/l). Subsequently, he developed supraventricular tachycardia, ventricular tachycardia and ultimately ventricular fibrillation resulting in cardiac arrest, which was successfully treated. A CT scan of the head revealed cerebral oedema. Whilst in the intensive care unit, he developed renal failure and rhabdomyolysis. The metabolic abnormalities seen at the time of admission normalised within 48 h with IV glucose infusion. Biological investigations, including urinary organic acids and plasma acylcarnitines, showed results compatible with MCAD deficiency. Mutation analysis revealed the patient was homozygous for the classical mutation A985G. This is one of only a few reports of severe cardiac arrhythmia in an adult due to MCAD deficiency. This condition is probably under-diagnosed in adult patients with acute neurological and/or cardiac presentations. PMID- 12897990 TI - Helium-oxygen reduces work of breathing in mechanically ventilated patients with chronic obstructive pulmonary disease. AB - OBJECTIVE: To evaluate whether helium-oxygen mixture reduces inspiratory work of breathing (WOB) in sedated, paralyzed, and mechanically ventilated patients with acute exacerbation of chronic obstructive pulmonary disease (COPD). DESIGN AND SETTING: Open, prospective, randomized, crossover study in the medical intensive care unit in a university hospital. PATIENTS AND PARTICIPANTS: 23 patients admitted for acute exacerbation of COPD and mechanically ventilated. MEASUREMENTS: Total WOB (WOBt), elastic WOB (WOBel), resistive WOB (WOBres), and WOB due to PEEPi (WOBPeepi) were measured. Static intrinsic positive end expiratory pressure (PEEPi), static compliance (Crs), inspiratory resistance (Rins), inspiratory (tinsp) and expiratory time constant (texp) were also measured. These variables were compared between air-oxygen and helium-oxygen mixtures. RESULTS: WOBt significantly decreased with helium-oxygen (2.34+/-1.04 to 1.85+/-1.01 J/l, p<0.001). This reduction was significant for WOBel (1.02+/ 0.61 J/l to 0.87+/-0.47, p<0.01), WOBPeepi (0.77+/-0.38 J/l to 0.54+/-0.38, p<0.001), and WOBres (0.55+/-0.19 J/l to 0.44+/-0.24, p<0.05). PEEPi, Rins, tinsp and texp significantly decreased. Crs was unchanged. CONCLUSIONS: Helium-oxygen mixture decreases WOB in mechanically ventilated COPD patients. Helium-oxygen mixture could be useful to reduce the burden of ventilation. PMID- 12897991 TI - Pulmonary glutamine production: effects of sepsis and pulmonary infiltrates. AB - OBJECTIVE: To define the role of the lung in the production of glutamine in the critically ill and to determine the effects of the presence of pulmonary infiltrates and the presence and severity of sepsis. DESIGN AND SETTING: Prospective clinical study in a single center; interdisciplinary intensive care unit at a university hospital. PATIENTS: Eleven critically ill patients were compared to ten patients prior to cardiac bypass surgery. MEASUREMENTS AND RESULTS: Fluxes of glutamine and other amino acids were measured. Chest radiography was performed, and APACHE II and multiple-organ failure scores were calculated. Septic patients showed significantly higher glutamine efflux from the lungs than controls. At least one-half of this glutamine is estimated to result from protein breakdown. Severity of illness had no impact on glutamine fluxes. In the presence of pulmonary infiltrates on chest radiographs glutamine efflux did not differ from zero. CONCLUSIONS: The lungs produce significant amounts of glutamine in septic patients. Pulmonary infiltrates decrease the glutamine efflux from the lung in septic patients. We suggest that this is caused by uptake of glutamine by white cells in the lung exerting immunological functions. PMID- 12897992 TI - Right atrial thrombus formation associated with central venous catheters utilization in hemodialysis patients. AB - OBJECTIVE: To report five patients on hemodialysis via an indwelling central venous catheter who developed a thrombus located exclusively on the right atrial wall opposing the emptying site of the superior vena cava and to determine the possible cause of this location. DESIGN: Transthoracic echocardiography was performed in four of the patients as work-up for suspected endocarditis or pulmonary embolism and in one patient for syncope evaluation. The right atrial clots were confirmed in all the patients by transesophageal echocardiography. DESIGN AND SETTING: General intensive care unit of a university hospital, tertiary referral center. PATIENTS: Five patients with end-stage chronic renal failure on hemodialysis via subclavian or internal jugular vein catheter. INTERVENTIONS: Three of these patients underwent surgical thrombectomy, and two others were medically treated. MEASUREMENTS AND RESULTS: The clots were 2-4 cm in length and three of them were infected. Two of the three surgically treated patients and one of the two medically treated patients died. All the patients had the catheter tip in the right atrium, in two of them the bent catheter rubbed the atrial endocardium, and in all the cases the clot was located on the atrial free wall facing the superior vena cava emptying. CONCLUSIONS: We postulate that the mechanism of thrombus formation at this location is related to friction of the catheter on the atrial endocardium, and therefore positioning the distal segment of the central venous catheters in the right atrium should be avoided. PMID- 12897993 TI - PEEP decreases atelectasis and extravascular lung water but not lung tissue volume in surfactant-washout lung injury. AB - OBJECTIVE: To examine the effects of positive end-expiratory pressure (PEEP) on extravascular lung water (EVLW), lung tissue, and lung volume. DESIGN AND SETTING: Experimental animal study at a university research facility. SUBJECTS: Fifteen adult sheep. INTERVENTIONS: All animals were studied before and after saline washout-induced lung injury while ventilated with sequentially increasing PEEP (0, 7, 14, or 21 cmH(2)O). MEASUREMENTS AND RESULTS: Lung volume was determined by computed tomography and EVLW by the thermal dye dilution technique. Saline washout significantly increased lung tissue volume (21+/-3 to 37+/-5 ml/kg) and EVLW (9+/-2 to 36+/-9 ml/kg). While increasing levels of PEEP reduced EVLW (30+/-7, 24+/-8, and 18+/-4 ml/kg), lung tissue volume remained constant. Total lung volume significantly increased (50+/-8 ml/kg at PEEP 0 to 77+/-12 ml/kg at PEEP 21). Nonaerated lung volume significantly decreased and was closely correlated with the changes in EVLW ( r=0.67). In addition, a highly significant correlation was found between PEEP-induced decrease in nonaerated lung volume and decrease in transpulmonary shunt ( r=0.83). CONCLUSIONS: The main findings are as follows: (a) PEEP effectively decreases EVLW. (b) The decrease in EVLW is closely correlated with the PEEP-induced decrease in nonaerated lung volume, making EVLW a valuable bedside parameter indicating alveolar recruitment, similar to measurements of transpulmonary shunt. (c) As excess tissue volume remained constant, however, EVLW may not be suitable to reflect overall severity of lung disease PMID- 12897994 TI - Omega-3 vs. omega-6 lipid emulsions exert differential influence on neutrophils in septic shock patients: impact on plasma fatty acids and lipid mediator generation. AB - OBJECTIVE: To compare the effects of a conventional omega-6 lipid infusion and a fish oil based (omega-3) lipid infusion for parenteral nutrition on neutrophil function, lipid mediators, and plasma free fatty acids. DESIGN AND SETTING: Open label, randomized, pilot study in a university hospital medical intensive care unit and experimental laboratory. PATIENTS AND PARTICIPANTS: Ten patients with septic shock and eight healthy controls. INTERVENTIONS: Patients (five per group) requiring parenteral nutrition received intravenously either a omega-3 or a omega 6 lipid emulsion for a 10-day period. MEASUREMENTS AND RESULTS: At baseline levels of plasma free fatty acids were elevated several-fold, including high concentrations of the omega-6 lipid precursor arachidonic acid (AA). Neutrophils isolated from septic patients displayed markedly reduced responsiveness to ex vivo stimulation, including lipid mediator generation [leukotrienes (LT), PAF], respiratory burst, and phosphoinositide hydrolysis signaling. Under the omega-6 lipid infusion regimen abnormalities in plasma free fatty acids and impairment of neutrophil functions persisted or worsened. In contrast, a rapid switch in the plasma free fatty acid fraction to predominance of the omega-3 acids eicosapentaenoic acid and docosahexaenoic acid over AA occurred in response to omega-3 lipid infusion. LTB(5), in addition to LTB(4), appeared upon neutrophil stimulation originating from these patients, and neutrophil function was significantly improved in the omega-3 lipid group. CONCLUSIONS: omega-3 vs. omega 6 lipid emulsions differentially influence the plasma free fatty acid profile with impact on neutrophil functions. Lipid-based parenteral nutrition in septic patients may thus exert profound influence on sequelae and status of immunocompetence and inflammation. PMID- 12897995 TI - Postoperative pain in the neonate: age-related differences in morphine requirements and metabolism. AB - OBJECTIVE: To investigate age-related differences in morphine requirements and metabolism in full-term neonates. DESIGN AND SETTING: Randomized double-blind study in the pediatric surgical intensive care unit. PATIENTS: Sixty-eight neonates (52 aged under 7 days, 16 aged 7 day or older) following major surgery. INTERVENTIONS: After surgery patients were randomly assigned to continuous morphine (10 micro g/kg per hour) or intermittent morphine (30 micro g/kg per 3 hours). Additional morphine was administered on guidance of pain scores. MEASUREMENTS AND RESULTS: Pain was measured by the Comfort behavioral scale and visual analogue scale. Morphine and morphine-6-glucuronide (M6G) plasma concentrations were determined before and 0, 6, 12, and 24 h after surgery. The younger neonates differed significantly from the older neonates in morphine requirement (median 10 vs. 10.8 micro g/kg per hour), morphine plasma concentration [23.0 vs. 15.3 ng/ml), and M6G/morphine ratio (0.6 vs. 1.5). Pain scores did not differ between age groups or morphine treatment groups. Neonates who were mechanically ventilated longer than 24 h had significantly higher morphine plasma concentrations than the spontaneously breathing neonates 12 and 24 h after surgery (29.1 vs. 13.1 ng/ml and 26.9 vs. 12.0 ng/ml, respectively). Morphine plasma concentrations were not correlated with analgesia or respiratory depression. Five neonates (intermittent morphine) showed respiratory insufficiency; however, the difference between the groups was not significant. CONCLUSIONS: Neonates aged 7 days or younger require significantly less morphine postoperatively than older neonates. The two morphine regimens (continuous, intermittent) were equally effective and safe. Mechanical ventilation decreases morphine metabolism and clearance. PMID- 12897996 TI - Gastric emptying in mechanically ventilated critically ill patients: effect of neuromuscular blocking agent. AB - OBJECTIVE: To assess gastrointestinal function in critically ill patients receiving muscle relaxant and to test clinical tolerance to enteral nutrition. DESIGN AND SETTING: Prospective study in an intensive care unit. PATIENTS: 20 critically ill patients requiring sedation with muscle relaxant to obtain adequate mechanical ventilation. MEASUREMENTS AND RESULTS: Patients were randomly selected to receive infusions of opioid sedation during the first session (session 1) and the same sedation with muscle relaxation (cisatracurium) during the second session (session 2). Gastric emptying was assessed by the paracetamol absorption technique. Following the paracetamol absorption 200 ml enteral feed was given, and the residual gastric volume was measured 1 and 2 h after feeding. The maximum plasma concentration (Cmax) was 14 mg/l (range 5-26) when patients received sedation, and 12 mg/l (range 5-30) when they received muscle relaxant. The target time for reaching the maximum plasma concentration (Tmax) was 30 min (range 20-60) and 35 min (range 20-60), respectively, in sessions 1 and 2. There was no significant difference between the two session as regards Tmax, Cmax, or AUC(0-120). The residual volumes were 110+/-65 ml (H1) and 95+/-76 ml (H2) during session 1 and 125+/-85 ml (H1) and 105+/-90 ml (H2) during session 2. CONCLUSIONS: Enteral feeding is one of the most effective methods of supporting nutritional needs in the critically ill patient. We conclude that in critically ill patients requiring sedation gastric emptying is not improved by neuromuscular blocking agent. PMID- 12897997 TI - High-frequency oscillatory ventilation in adult acute respiratory distress syndrome. AB - OBJECTIVE: This study examined whether ARDS patients in whom predefined ventilator settings fail to maintain oxygenation and CO2 removal can be safely transitioned to high-frequency oscillatory ventilation (HFOV), and whether HFOV use is efficacious. DESIGN AND SETTING: Prospective observational study in the 14 bed intensive care unit of a university hospital. PATIENTS AND PARTICIPANTS: 42 patients with ARDS (APACHE II score 28 (IQR 24-37) and ventilation time prior HFOV 3.0 days (0.7-9.1). MEASUREMENTS AND RESULTS: Gas exchange parameters and ventilator data were recorded before and during HFOV treatment (-12 h, -6 h, baseline, 10 min, 1 h, 6 h, 12 h, 24 h). Primary endpoints included: (a) PaO2/FIO2 ratio 24 h after start of HFOV treatment or the last point of measurement if HFOV ended within the first 24 h; (b) HFOV-related complications. Post hoc analysis assessed the relationship between outcome and the response to HFOV, and between outcome and time of mechanical ventilation prior to HFOV. At baseline the median PaO2/FIO2 ratio was 95 (IQR 62-129); after 24 h of HFOV the PaO2/FIO2 ratio had increased significantly to 165 (88-225); only one patient developed a unilateral pneumothorax. Of the 42 patients 18 (43%) had died by day 30. Subset analyses showed a significantly higher 30-day mortality rate in patients with at least 3 days of mechanical ventilation prior to HFOV (64%) and in patients without oxygenation improvement after 24 h on HFOV (71%). CONCLUSIONS: HFOV is an effective and safe method to ventilate ARDS patients. Failure to improve oxygenation within 24 h of HFOV is associated with high mortality. PMID- 12897998 TI - Effect of forced deflation maneuvers upon measurements of respiratory mechanics in ventilated infants. AB - OBJECTIVE: To determine the effect of forced deflation maneuvers on respiratory mechanics and to assess the reproducibility of such measurements in intubated infants with lung disease. DESIGN AND SETTING: Prospective study in the pediatric intensive care unit of a university children's hospital. PATIENTS: Ten clinically stable infants requiring mechanically assisted ventilation for acute pulmonary disease, mean age 5.9 months (1-18), mean weight 5.8 kg (3.2-13). INTERVENTIONS: Two sets of measurements of compliance (Crs) and resistance (Rrs) were obtained at 20-min intervals both before and after +40/-40 cmH(2)O forced deflation maneuvers. Forced deflation measurements were repeated at the end of the study. RESULTS: . Forced deflation caused a significant increase in Crs from 0.53+/-0.09 and 0.58+/-0.11 ml/cmH(2)O/kg to 0.71+/-0.11 and 0.68+/-0.11 ml/cmH(2)O/kg. Rrs measurements did not differ. The low coefficients of variation for repeated measures of the baseline measurements (Crs 4.2+/-0.5%, Rrs 7.1+/-0.8%, for forced vital capacity 8.6+/-2.5%, maximum expiratory flows at 25% vital capacity 16.0%+/ 3.3%) confirmed the good reproducibility during stable conditions. CONCLUSIONS: Inflation and deflation maneuvers affect subsequent measurements of respiratory system compliance but not measurements of maximum expiratory flow-volume relationships in intubated infants, probably through recruitment of lung volume. Careful interpretation and planning of the sequence of infant pulmonary function testing is necessary to reassure that changes are not related to short-term alterations in volume history. PMID- 12897999 TI - Transport of critically ill children in a resource-limited setting. AB - OBJECTIVE: To audit paediatric intensive care unit (PICU) transfer activity and transfer-related adverse events in a resource-limited setting. DESIGN AND SETTING: Twenty-two bed regional PICU of a university children's hospital in Cape Town, South Africa. Prospective 1-year audit of all children transferred directly from other hospitals. Data were collected for patient demographics, diagnostic category, referring hospital, transferring personnel, mode of transport, and technical, clinical, and critical adverse events. Data are median (interquartile range) or percentages. The transfers of 202 children, median age 2.8 months (1.1 14), median weight 3.5 kg (2.5-8.1) were analysed. RESULTS: Most transfers were performed by paramedic personnel (82%) and via road ambulance (76%). One or more technical adverse events occurred in 36%, clinical adverse events in 27%, and critical adverse events in 9% of children. Retrievals by intensive care staff (10%), all from rural hospitals, had a lower incidence of technical adverse events (0%). Children transferred from non-academic hospitals within the metropolitan area had the highest incidence of technical (44%), clinical (39%), and critical (17%) adverse events. Crude mortality was 17% ( n=34). Technical adverse events were not associated with mortality. Non-survivors were more likely to develop shock (32%) or hypoxia (26%) during transfer than survivors (10% and 11%, respectively). CONCLUSIONS: There is a high incidence of transfer-related adverse events, most commonly in transfers from non-academic metropolitan hospitals. Further studies are needed to assess the impact of regional paediatric life support training or a specialised retrieval team on clinical adverse events and mortality. PMID- 12898000 TI - Doppler-guided cannulation of internal jugular vein, subclavian vein and innominate (brachiocephalic) vein--a case-control comparison in patients with reduced and normal intracranial compliance. AB - OBJECTIVE: A case-control comparison of Doppler guidance on the success rate of central venous cannulation in patients with normal or reduced intracranial compliance. DESIGN: A single operator performed central venous access procedures with continuous wave Doppler guidance. It was used on patients on a ventilator. The position of patients with reduced intracranial compliance (RIC) was not changed for the procedure. Patients with normal intracranial compliance (NIC) were put in the Trendelenburg position. SETTING: We prospectively evaluated 249 Doppler-guided central venous access procedures performed over a 12-month period at our 10-bed neuro-intensive care unit at a university hospital. PATIENTS AND PARTICIPANTS: The group with RIC included 26 males and 35 females (n=61) aged 16 79 years. In this group 155 Doppler-guided cannulation procedures (62%) were performed. The group with NIC (n=52) comprised 29 males and 23 females aged 34-76 years; 94 Doppler-guided cannulation procedures (38%) were carried out. MEASUREMENTS AND RESULTS: The veins cannulated in RIC and NIC, respectively, were: right innominate vein: 24/18, left innominate vein 26/12, right subclavian vein 12/7, left subclavian vein 25/14, and right internal jugular vein 33/18 and left internal jugular vein 35/24. The absence of one left internal jugular vein was identified in the NIC group. The success rate of first needle pass in patients with RIC was 92% and in patients with NIC 89%. CONCLUSIONS: This study showed that Doppler guidance allows the cannulation of central veins in patients with RIC placed in head-up position. Cannulation can be ensured and first-pass needle placement maximised. PMID- 12898001 TI - Explicit approach to rounds in an ICU improves communication and satisfaction of providers. AB - OBJECTIVE: To evaluate an explicit approach to responsibilities and reporting during bedside rounds in the intensive care unit (ICU). DESIGN AND SETTING: Before-after comparison in the 15-bed medical-surgical ICU in a tertiary teaching hospital. PATIENTS: All patients in the ICU during two 1-month periods: 2 months before and 16 months after the intervention. INTERVENTIONS: Explicit approach to clinical and educational responsibilities and to reporting assessments and plans during bedside rounds. MEASUREMENTS AND RESULTS: Surveys of all ICU staff (attending physicians, residents, nurses, respiratory therapists, pharmacists, and medical students) were conducted during rounds on each patient. After the intervention, more respondents agreed that there was a long-term plan for each patient (74% vs. 53% before), that the long-term plan was clear (76% vs. 54% before), that there was structured teaching around each patient (46% vs. 30% before), that discussions other than structured teaching around each patient were useful (79% vs. 65% before), and that they were satisfied with the process and outcome of rounds (95% vs. 86% before). In contrast, slightly fewer respondents were aware of a problem list for the patients (96% vs. 99% before), and fewer residents and medical students had examined their patients before rounds (76% vs. 88% before). CONCLUSIONS: An explicit approach to bedside rounds in an ICU improves communication and satisfaction of health care providers. PMID- 12898003 TI - Life-threatening snakebites by Vipera berus. PMID- 12898002 TI - Nosocomial infections in intensive care unit in a Turkish university hospital: a 2-year survey. AB - OBJECTIVE: To determine epidemiology and risk factors for nosocomial infections in intensive care unit (ICU). DESIGN. Prospective incidence survey. SETTING: An adult general ICU in a university hospital in western Turkey. PATIENTS: All patients who stayed more than 48 h in ICU during a 2-year period (2000-2001). MEASUREMENTS AND RESULTS: The study included 434 patients (7394 patient-days). A total of 225 infections were identified in 113 patients (26%). The incidence and infection rates were 56.8 in 1000-patient days and 51.8%, respectively. The infections were pneumonia (40.9%), bloodstream (30.2%), urinary tract (23.6%) and surgical site infections (5.3%). Pseudomonas aeruginosa (22.6%), methicillin resistant Staphylococcus aureus (22.2%) and Acinetobacter spp. (11.9%) were frequently isolated micro-organisms. Median length of stay with nosocomial infection and without were 13 days (Interquartile range, IQR, 20) and 2 days (IQR, 2), respectively ( P<0.0001). In logistic regression analysis, mechanical ventilation [odds ratio (OR): 16.35; 95% confidence interval (CI): 8.26-32.34; P<0.0001), coma (OR: 15.04; 95% CI: 3.41-66.33; P=0.0003), trauma (OR: 10.27; 95% CI: 2.34-45.01; P=0.002), nasogastric tube (OR: 2.94; 95% CI: 1.47-5.90; P=0.002), tracheotomy (OR: 5.77; 95% CI: 1.10-30.20; P=0.04) and APACHE II scores 10-19 (OR: 10.80; 95% CI: 1.10-106.01; P=0.04) were found to be significant risk factors for nosocomial infection. Rate of nosocomial infection increased with the number of risk factors (P<0.0001). Mortality rates were higher in infected patients than in non-infected patients (60.9 vs 22.1%; P<0.0001). CONCLUSION: These data suggest that, in addition to underlying clinical conditions, some invasive procedures can be independent risk factors for nosocomial infection in ICU. PMID- 12898004 TI - Three generations of autoimmune diabetes: an extended family study. AB - AIMS/HYPOTHESIS: One in four children with Type 1 diabetes in a population-based family study has an affected grandparent. We set out to study the clinical and immune features of diabetes in the grandparents' generation, and to examine sharing of HLA class II susceptibility haplotypes between grandparent and grandchild. METHODS: Of 5855 grandparents in the Bart's-Oxford family study, 428 (7.3%) were known to have diabetes. Clinical data and samples were collected from 115 of 213 surviving affected grandparents and from 219 unaffected grandparents within the same families. Samples were tested for ICA and autoantibodies to GAD and IA-2, and typed for HLA-DRB1-DQA1-DQB1. Transmission of HLA class II haplotype from affected and unaffected grandparents to the diabetic proband was compared. RESULTS: Of 115 affected grandparents studied, the median age at diagnosis was 61 years and at analysis was 73 years; 70% were diet or tablet treated and 30% were on insulin. One or more islet autoantibodies were found in 26% and 66% had one or both of the high risk HLA class II susceptibility haplotypes DRB1*03-DQA1*0501-DQB1*0201 or DRB1*04-DQA1*0301-DQB1*0302. In 79 informative families the HLA class II haplotype of the affected grandparent was transmitted to the proband more frequently than expected overall (59%, p=0.02), and in the insulin-treated subgroups (65%, p=0.03). CONCLUSION/INTERPRETATION: A total of 7.3% of grandparents reported a clinical diagnosis of diabetes and 2.2% had features of Type 1 diabetes. Genetic susceptibility was shared between grandparents with diabetes and their affected grandchildren. Diabetes in the grandparents of children with Type 1 diabetes often has an autoimmune basis, even when it presents late in life and does not require insulin treatment. PMID- 12898005 TI - Saturated-fat diet induces moderate diabetes and severe glomerulosclerosis in hamsters. AB - AIMS/HYPOTHESIS: Diets high in saturated fat are thought to be a risk factor for Type 2 diabetes and associated complications. We investigated effects of a medium and high saturated fat diet on the development of diabetes-associated pathologies in Golden Syrian hamsters, an animal that reacts to dietary lipids in a fashion similar to humans. METHODS: Three diets containing 46.5 kcal %, 267.3 kcal %, and 488.2 kcal % as saturated fat respectively, were studied. Metabolic parameters were measured up to 20 weeks. Electron microscopy was used to examine the structure of the pancreas, aorta and kidney. RESULTS: Increased saturated fat consumption was associated with: (i) gradual imbalance of homeostasis, and severe structural alterations of acinar, beta cells and capillaries in the pancreas, and of the kidney glomeruli; (ii) severe hypertriglyceridaemia and augmented creatinine concentrations related to disturbances of the renal function, progressing to nodular glomerulosclerosis and nephropathy; (iii) reduced early insulin secretion in response to glucose; (iv) switch of the aortic endothelium to a secretory phenotype. CONCLUSION/INTERPRETATION: The results show that high caloric saturated fat intake induced diabetes in hamsters, probably linked to delayed insulin secretion. The model was also associated with the development of a range of pathologies characteristic to human diabetes, including nephropathy and defects in vasculature. Thus, high-fat fed hamsters provide a new model that is likely to be useful in understanding the cellular and molecular mechanisms involved in the pathogenesis of diabetes. PMID- 12898006 TI - Little evidence of transdifferentiation of bone marrow-derived cells into pancreatic beta cells. AB - AIMS/HYPOTHESIS: Bone marrow cells contain at least two distinct types of stem cells which are haematopoietic stem cells and mesenchymal stem cells. Both cells have the ability to differentiate into a variety of cell types derived from all three germ layers. Thus, bone marrow stem cells could possibly be used to generate new pancreatic beta cells for the treatment of diabetes. In this study, we investigated the feasibility of bone marrow-derived cells to differentiate into beta cells in pancreas. METHODS: Using green fluorescent protein transgenic mice as donors, the distribution of haematogenous cells in the pancreas was studied after bone marrow transplantation. RESULTS: In the pancreas of green fluorescent protein chimeric mice, green fluorescent protein-positive cells were found in the islets, but none of these cells expressed insulin. Previous data has suggested that tissue injury can recruit haematopoietic stem cells or their progeny to a non-haematopietic cell fate. Therefore, low-dose streptozotocin (30 or 50 mg/kg on five consecutive days) was injected into the mice 5 weeks after bone marrow transplantation, but no green fluorescent protein-positive cells expressing insulin were seen in the islets or around the ducts of the pancreas. CONCLUSIONS/INTERPRETATION: Our data suggests that bone marrow-derived cells are a distinct cell population from islet cells and that transdifferentiation from bone marrow-derived cells to pancreatic beta cells is rarely observed. PMID- 12898007 TI - Gemfibrozil considerably increases the plasma concentrations of rosiglitazone. AB - AIMS/HYPOTHESIS: Our aim was to investigate possible interaction between gemfibrozil and rosiglitazone, a thiazolidinedione antidiabetic drug. METHODS: In a randomised crossover study with two phases, 10 healthy volunteers took 600 mg gemfibrozil or placebo orally twice daily for 4 days. On day 3, they ingested a single 4 mg dose of rosiglitazone. Plasma rosiglitazone and its N-desmethyl metabolite concentrations were measured for up to 48 h. RESULTS: Gemfibrozil raised the mean area under the plasma rosiglitazone concentration-time curve (AUC) 2.3-fold (range 1.5- to 2.8-fold; p=0.00002) and prolonged the elimination half-life (t(1/2)) of rosiglitazone from 3.6 to 7.6 h ( p=0.000002). The peak plasma rosiglitazone concentration (C(max)) was increased only 1.2-fold (range 0.9- to 1.6-fold; p=0.01) by gemfibrozil, but gemfibrozil raised the plasma rosiglitazone concentration measured 24 h after dosing (C(24)) 9.8-fold (range, 4.5- to 33.6-fold; p=0.00008). In addition, gemfibrozil prolonged the t(max) of N desmethylrosiglitazone from 7 to 12 h and reduced the N desmethylrosiglitazone/rosiglitazone AUC(0-48) ratio by 38% ( p<0.01). CONCLUSIONS/INTERPRETATION: Gemfibrozil raises the plasma concentrations of rosiglitazone probably by inhibiting the CYP2C8-mediated biotransformation of rosiglitazone. Co-administration of gemfibrozil, or another potent inhibitor of CYP2C8, and rosiglitazone could increase the efficacy but also the risk of concentration-dependent adverse effects of rosiglitazone. PMID- 12898008 TI - Peripheral insulin resistance develops in transgenic rats overexpressing phosphoenolpyruvate carboxykinase in the kidney. AB - AIMS/HYPOTHESIS: To study the secondary consequences of impaired suppression of endogenous glucose production (EGP) we have created a transgenic rat overexpressing the gluconeogenic enzyme phosphoenolpyruvate carboxykinase (PEPCK) in the kidney. The aim of this study was to determine whether peripheral insulin resistance develops in these transgenic rats. METHODS: Whole body rate of glucose disappearance (R(d)) and endogenous glucose production were measured basally and during a euglycaemic/hyperinsulinaemic clamp in phosphoenolpyruvate carboxykinase transgenic and control rats using [6-(3)H]-glucose. Glucose uptake into individual tissues was measured in vivo using 2-[1-(14)C]-deoxyglucose. RESULTS: Phosphoenolpyruvate carboxykinase transgenic rats were heavier and had increased gonadal and infrarenal fat pad weights. Under basal conditions, endogenous glucose production was similar in phosphoenolpyruvate carboxykinase transgenic and control rats (37.4+/-1.1 vs 34.6+/-2.6 micromol/kg/min). Moderate hyperinsulinaemia (810 pmol/l) completely suppressed EGP in control rats (-0.6+/ 5.5 micromol/kg/min, p<0.05) while there was no suppression in phosphoenolpyruvate carboxykinase rats (45.2+/-7.9 micromol/kg/min). Basal R(d) was comparable between PEPCK transgenic and control rats (37.4+/-1.1 vs 34.6+/ 2.6 micromol/kg/min) but under insulin-stimulated conditions the increase in R(d) was greater in control compared to phosphoenolpyruvate carboxykinase transgenic rats indicative of insulin resistance (73.4+/-11.2 vs 112.0+/-8.0 micromol/kg/min, p<0.05). Basal glucose uptake was reduced in white and brown adipose tissue, heart and soleus while insulin-stimulated transport was reduced in white and brown adipose tissue, white quadriceps, white gastrocnemius and soleus in phosphoenolpyruvate carboxykinase transgenic compared to control rats. The impairment in both white and brown adipose tissue glucose uptake in phosphoenolpyruvate carboxykinase transgenic rats was associated with a decrease in GLUT4 protein content. In contrast, muscle GLUT4 protein, triglyceride and long-chain acylCoA levels were comparable between PEPCK transgenic and control rats. CONCLUSIONS/INTERPRETATION: A primary defect in suppression of EGP caused adipose tissue and muscle insulin resistance. PMID- 12898009 TI - Diabetes-induced activation of protein kinase C inhibits store-operated Ca2+ uptake in rat retinal microvascular smooth muscle. AB - AIMS/HYPOTHESIS: To assess the effects of diabetes-induced activation of protein kinase C (PKC) on voltage-dependent and voltage-independent Ca2+ influx pathways in retinal microvascular smooth muscle cells. METHODS: Cytosolic Ca2+ was estimated in freshly isolated rat retinal arterioles from streptozotocin-induced diabetic and non-diabetic rats using fura-2 microfluorimetry. Voltage-dependent Ca2+ influx was tested by measuring rises in [Ca2+]i with KCl (100 mmol/l) and store-operated Ca2+ influx was assessed by depleting [Ca2+]i stores with Ca2+ free medium containing 5 micromol/l cyclopiazonic acid over 10 min and subsequently measuring the rate of rise in Ca2+ on adding 2 mmol/l or 10 mmol/l Ca2+ solution. RESULTS: Ca2+ entry through voltage-dependent L-type Ca2+ channels was unaffected by diabetes. In contrast, store-operated Ca2+ influx was attenuated. In microvessels from non-diabetic rats 20 mmol/l D-mannitol had no effect on store-operated Ca2+ influx. Diabetic rats injected daily with insulin had store-operated Ca2+ influx rates similar to non-diabetic control rats. The reduced Ca2+ entry in diabetic microvessels was reversed by 2-h exposure to 100 nmol/l staurosporine, a non-specific PKC antagonist and was mimicked in microvessels from non-diabetic rats by 10-min exposure to the PKC activator phorbol myristate acetate (100 nmol/l). The specific PKCbeta antagonist LY379196 (100 nmol/l) also reversed the poor Ca2+ influx although its action was less efficacious than staurosporine. CONCLUSION/INTERPRETATION: These results show that store-operated Ca2+ influx is inhibited in retinal arterioles from rats having sustained increased blood glucose and that PKCbeta seems to play a role in mediating this effect. PMID- 12898010 TI - The effects of a specific growth hormone antagonist on overnight insulin requirements and insulin sensitivity in young adults with Type 1 diabetes mellitus. AB - AIMS/HYPOTHESIS: Growth hormone hypersecretion in Type 1 diabetes could exacerbate insulin resistance and contribute to declining glycaemic control. Our aim was to determine the effects of specific growth hormone blockade on insulin sensitivity and lipolysis in young adults with Type 1 diabetes. METHODS: We studied the effects of two doses of a specific growth hormone antagonist (B2036 PEG; Somavert, Pharmacia Corporation, Milton Keynes, UK) on insulin sensitivity in seven young adults (17-22 yrs, 3M) with Type 1 diabetes. Subjects received 5 and 10 mg B2036-PEG, in random order for 3 weeks, with a 3-week washout. At baseline and following each treatment block, an overnight (03:00 to 08:00 h) insulin infusion for euglycaemia (5 mmol/l), followed by two-step hyperinsulinaemic euglycaemic clamp, using [6,6 2H2] glucose and 2H5 glycerol to measure glucose and glycerol turnover was performed. RESULTS: Compared to baseline, overnight insulin requirements decreased with both doses: (means+/-SEM) 0.34+/-0.02 mU/Kg/min vs 0.25+/-0.01 (5 mg) (p=0.04), and 0.24+/-0.01 (10 mg) (p=0.004). IGF-I (ng/ml) decreased following 10 mg [223.5+/-23.9 vs 154.6+/-28.1 (p=0.005], but not 5 mg. Mean overnight non esterified fatty acid concentrations (mmol/l) decreased with 10 mg [0.51+/-0.04 vs 0.38+/-0.04 (p=0.03)], as did beta hydroxybutyrate (mmol/l); [0.31+/-0.04 vs 0.15+/-0.02 (p=0.004)]. Glycerol production rate, an index of lipolysis, was lower following 10 mg (p=0.04), but insulin sensitivity during the clamp did not change with either dose. CONCLUSION/INTERPRETATION: Treatment with both doses of B2036-PEG reduced overnight insulin requirements. The 10 mg dose suppressed lipolysis and reduced IGF-I. Failure to show enhanced insulin sensitivity during the clamp with the 10 mg dose could reflect opposing actions of growth hormone and IGF-I. PMID- 12898011 TI - Haeme-oxygenase 1 expression in rat pancreatic beta cells is stimulated by supraphysiological glucose concentrations and by cyclic AMP. AB - AIM/HYPOTHESIS: Increased expression of haeme-oxygenase 1 (HO1) and other antioxidant enzymes could improve pancreatic beta-cell survival under stressful conditions, including hyperglycaemia. However, how hyperglycaemia increases islet HO1 expression is not known. METHODS: Rat islets were pre-cultured for 1 week in RPMI medium containing 10 mmol x l(-1) glucose (G10), and further cultured overnight in G5-G30 plus various test substances. Islet HO1 mRNA and protein expression was measured by semiquantitative RT-PCR, western blot, and immunohistochemistry. RESULTS: Islet HO1 mRNA expression was minimal after overnight culture in G10, slightly increased in G5, and increased by five- to ten fold in G30 in parallel with a heterogeneous increase in beta-cell HO1 protein expression. The effect of G30 was fully inhibited by agents decreasing cytosolic Ca2+ (diazoxide, nimodipine), but was only slightly reproduced by agents raising Ca2+ (tolbutamide, 30 mmol x l(-1) potassium). It was also suppressed by the alpha2-adrenoceptor agonist clonidine, whereas dibutyryl-cyclic-AMP largely increased beta-cell HO1 expression. The induction of HO1 mRNA expression by G30 was independent from changes in medium insulin concentration, but was completely inhibited by a cocktail of antioxidants. In contrast to HO1, islet mRNA expression of glutathione peroxidase and constitutive haeme-oxygenase 2 were not affected by G30, nor by dibutyryl-cyclic-AMP. CONCLUSION/INTERPRETATION: High glucose and dibutyryl-cyclic-AMP stimulate expression of HO1 in rat pancreatic beta cells. The inhibition of HO1 expression in G30 by nimodipine, clonidine, and antioxidants, suggests that Ca2+ influx and cyclic-AMP are necessary for the generation of oxidative stress by G30, or for the stimulation of beta-cell HO1 expression by increased oxidative stress. PMID- 12898012 TI - Suppressive effects of a selective inducible nitric oxide synthase (iNOS) inhibitor on pancreatic beta-cell dysfunction. AB - AIMS/HYPOTHESIS: Type 1 diabetes mellitus is an autoimmune disease characterized by dysfunction and destruction of the pancreatic beta cells. Interleukin-1beta (IL-1beta) has been reported to cause suppression of insulin secretion from pancreatic islets via induction of inducible nitric oxide synthase (iNOS) followed by nitric oxide (NO) production. In this study, we investigated the effects of inhibition of iNOS on pancreatic beta-cell dysfunction in non-obese diabetic (NOD) mice and IL-1beta-treated isolated rat pancreatic islets using a novel specific inhibitor, ONO-1714. METHODS: Female NOD mice which received subcutaneous infusion of ONO-1714 (4 microg/kg/day or 40 microg/kg/day) from 10 to 14 weeks after birth were compared with untreated NOD mice. In addition, pancreatic islets were isolated from Sprague-Dawley rats and cultured for 24 h with IL-1beta (100 U/mL) with or without ONO-1714 or the non-selective NOS inhibitor NG-monomethyl-L-arginine (L-NMMA). We measured insulin secretion and insulin content of the islets by ELISA, iNOS mRNA expression by reverse transcription-polymerase chain reaction, and NO generation by Griess Reagent System. RESULTS: Hyperglycaemia was observed in NOD mice. ONO-1714 treatment blunted this increase and tended to preserve insulin secretion, although body weight increase did not differ between the groups. Insulitis was also attenuated in the ONO-1714-administered group compared to the control group. Furthermore, in isolated rat pancreatic islets ONO-1714 prevented IL-1beta-induced inhibition of insulin secretion, this protection being evident in much lower concentrations than with L-NMMA. While ONO-1714 completely inhibited IL-1beta-induced NO production, it did not reduce expression of islet iNOS mRNA. CONCLUSION/INTERPRETATION: These findings indicate that ONO-1714 is promising as a therapeutic agent for autoimmune diabetes. PMID- 12898013 TI - Pulmonary vasodilation in the rat by insulin in vitro could indicate potential hazard for inhaled insulin. AB - AIMS/HYPOTHESIS: Hypoxic pulmonary vasoconstriction is an essential mechanism to prevent hypoxaemia in lung diseases. Insulin is known to be a systemic vasodilator but its effect on the pulmonary circulation is not known. Inhaled particulate insulin can generate locally high concentrations in the lung which could be physiologically important. METHODS: We therefore studied the effects of insulin in vitro on isolated rat pulmonary artery in a small vessel myograph. RESULTS: We have shown that pulmonary artery vasodilatation with insulin occurs in a dose-dependent manner. Pre-constriction with PGF2alpha can be abolished (105.7+/-2.9%, mean+/-SEM) and pre-constriction with hypoxia reduced (68.9+/ 6.5%) by pharmacologically relevant concentrations of insulin. The characteristic phasic vasoconstriction by pulmonary vessel to hypoxia is substantially modified, resulting in sustained vasodilatation. CONCLUSIONS/INTERPRETATION: These effects could be clinically important for patients using inhaled insulins who have acute or occult chronic lung disease. PMID- 12898014 TI - Twin study of genetic and environmental influences on glucose tolerance and indices of insulin sensitivity and secretion. AB - AIMS/HYPOTHESIS: Family and twin studies have reported different estimates of the relative contribution of genetic and environmental factors to the quantitative traits glucose tolerance, insulin secretion, and insulin sensitivity. Our aims were to estimate these relative influences in a large sample of twins from the population and to assess the effect of age. METHODS: In this population-based, cross-sectional study we gave an oral glucose tolerance test to 317 women and 290 men who were same-sex healthy twin pairs between 18 to 67 years of age. The genetic, common environmental and individual environmental variance components for fasting and 120-min glucose and for fasting and 30-min insulin as well as the linear effects of age on these components were estimated by multivariate analysis (using the software FISHER). RESULTS: In women and men the heritability for fasting glucose was 12 and 38%, for 120-min glucose it was 38 and 43%, for fasting insulin it was 54 and 37%, and for 30-min insulin it was 57 and 47%, respectively. Under the assumption of no non-additive genetic effects (no intra- or inter-gene interaction) there was no strong evidence for common environmental effects, barring significant effects for fasting glucose in women. Heritability decreased with age for 120-min glucose in women and fasting insulin in men, whereas it increased for 120-min glucose in men. CONCLUSION/INTERPRETATION: This study indicates a limited additive genetic influence on the result of an OGTT, possibly with sex-specific age effects, and generally little or no influence of the common environment. Accordingly, there is a considerable individual environmental variation. PMID- 12898015 TI - Role of nitric oxide, tetrahydrobiopterin and peroxynitrite in glucose toxicity associated contractile dysfunction in ventricular myocytes. AB - AIMS/HYPOTHESIS: Local overproduction of nitric oxide is seen in early stages of diabetes, which can react with superoxide (O(2)(-)) to form peroxynitrite (ONOO( )). The aim of this study was to examine the effect of scavengers for nitric oxide, O(2)(-), ONOO(-) and NOS cofactor tetrahydrobiopterin (BH(4)) on high glucose-induced cardiac contractile dysfunction. METHODS: Ventricular myocytes were cultured for 24 h with either normal (N, 5.5 mmol/l) or high (25.5 mmol/l) glucose, with or without the nitric oxide scavengers haemoglobin (100 nmol/l), PTIO (100 micromol/l), the NOS inhibitor L-NMMA (100 micromol/l), superoxide dismutase (SOD, 500 U/ml), the ONOO(-) scavengers urate (100 micromol/l), MnTABP (100 micromol/l), BH(4) (10 micromol/l) and its inactive analogue NH(4) (10 micromol/l), and the GTP cyclohydrolase I inhibitor DAHP (1 mmol/l). Myocyte mechanics, NOS protein expression and activity were evaluated. RESULTS: High glucose myocytes showed reduced peak shortening, decreased maximal velocity of shortening/relengthening (+/- dL/dt), prolonged relengthening (TR(90)) and normal shortening duration (TPS) associated with reduced cytosolic Ca(2+) rise compared to normal myocytes. The high glucose-induced abnormalities were abrogated or attenuated by urate, MnTBAP, L-NMMA, BH(4), and SOD, whereas unaffected by haemoglobin, PTIO and NH(4). L-NMMA reduced peak shortening while PTIO and DAHP depressed +/- dL/dt and prolonged TPS or TR(90) in normal myocytes. High glucose increased NOS activity, protein expression of eNOS but not iNOS, which were attenuated by L-NMMA and BH(4), respectively. CONCLUSION/INTERPRETATION: These results suggested that NOS cofactor, NO and ONOO(-) play a role in glucose induced cardiomyocyte contractile dysfunction and in the pathogenesis of diabetic cardiomyopathy. PMID- 12898016 TI - Type 1 diabetes islet associated antibodies in subjects infected by echovirus 16. AB - AIMS/HYPOTHESIS: To determine whether the emergent infection by echovirus 16 that occurred in Cuba during the year 2000 was related to the presence of Type 1 diabetes associated autoantibodies. METHODS: The presence of ICA, IAA, GADA, IA2 antibodies and neutralizing antibodies (NtAb) to echovirus 16 were determined in sera from 38 infected children and adolescents and 80 control subjects, matched in sex, age, local residence and time of sample collection. RESULTS: The occurrence of a large-scale echovirus 16 epidemic was associated with the appearance of humoral autoimmune markers of Type 1 diabetes, especially for ICA, IAA and GADA. In the convalescent stage, ICA, IAA and GADA seroconversion was shown in 92.1%, 44.7% and 28.9% of echovirus 16 infected subjects. None of the 80 uninfected subjects had ICA or IAA, while one was GADA positive. ICA, IAA and GADA frequency was higher in the convalescent than in the acute stage (p<0.0005). A strong positive correlation was found between the NtAb to echovirus 16 and ICA titres in both acute and convalescent stage (r=0.91; p<0.0001, r=0.55; p=0.0003 respectively). CONCLUSION/INTERPRETATION: This work provides evidence of an association between echovirus 16 infection and the presence of Type 1 diabetes related antibodies (ICA, IAA and GADA). Our data show that the echovirus 16 infection might be capable of inducing a process of autoimmune beta-cell damage and support the hypothesis that enterovirus infections are important risk factors for the development of Type 1 diabetes. PMID- 12898017 TI - Apoptotic death of photoreceptors in the streptozotocin-induced diabetic rat retina. AB - AIMS/HYPOTHESIS: Neurodegenerative changes in the diabetic retina occurring before diabetic retinopathy could be inevitable by the altered energy (glucose) metabolism, in the sense that dynamic image-processing activity of the retinal neurons is exclusively dependent on glucose. We therefore investigated the morphological changes in the neural retina, including neuronal cell death, of a streptozotocin-induced model of diabetes. METHODS: Streptozotocin was intravenously injected. Rats were maintained hyperglycaemic without insulin treatment for 1 week and 4, 8, 12, and 24 weeks, respectively. Diabetic retinas were processed for histology, electron microscopy, and immunohistochemistry using the TUNEL method. RESULTS: A slight reduction in the thickness of the inner retina was observed throughout the diabetic retinas and a remarkable reduction was seen in the outer nuclear layer 24 weeks after the onset of diabetes. The post-synaptic processes of horizontal cells in the deep invaginations of the photoreceptors showed degeneration changes from 1 week onwards. A few necrotic ganglion cells were observed after 4 weeks. At 12 weeks, some amacrine cells and a few horizontal cells showed necrotic features. Three to seven cellular layers in the outer nuclear layer and nerve terminals, rolled by the fine processes of the Muller cells near the somata of the degenerated ganglion cells, were apparent at 24 weeks. Apoptosis appeared in a few photoreceptor cells at 4 weeks, and the number of apoptotic photoreceptors increased thereafter. CONCLUSION/INTERPRETATION: These findings suggest that the visual loss associated with diabetic retinopathy could be attributed to an early phase of substantial photoreceptor loss, in addition to later microangiopathy. PMID- 12898019 TI - BIBAC and TAC clones containing potato genomic DNA fragments larger than 100 kb are not stable in Agrobacterium. AB - Development of efficient methods to transfer large DNA fragments into plants will greatly facilitate the map-based cloning of genes. The recently developed BIBAC and TAC vectors have shown potential to deliver large DNA fragments into plants via Agrobacterium-mediated transformation. Here we report that BIBAC and TAC clones containing potato genomic DNA fragments larger than 100 kb are not stable in Agrobacterium. We tested the possible factors that may cause instability, including the insert sizes of the BIBAC and TAC constructs, potato DNA fragments consisting of highly repetitive or largely single-copy DNA sequences, different Agrobacterium transformation methods and different Agrobacterium strains. The insert sizes of the potato BIBAC and TAC constructs were found to be critical to their stability in Agrobacterium. All constructs containing a potato DNA fragment larger than 100 kb were not stable in any of the four tested Agrobacterium strains, including two recA deficient strains. We developed a transposon-based technique that can be used to efficiently subclone a BAC insert into two to three BIBAC/TAC constructs to circumvent the instability problem. PMID- 12898018 TI - Pyruvate inhibits zinc-mediated pancreatic islet cell death and diabetes. AB - AIMS/HYPOTHESIS: We have shown that zinc ion (Zn2+) in secretory granules of pancreatic beta cells could act as a paracrine death effector in streptozotocin induced diabetes. As Zn2+ has been reported to perturb glycolysis, we studied if pyruvate could inhibit Zn(2+)-mediated islet cell death in vitro and streptozotocin-induced diabetes in vivo by normalizing intracellular energy metabolism. METHODS: Cell death was measured by quantitative viable cell staining and Hoechst/propidium iodide staining. ATP was measured by bioluminescence determination. Pyruvate was infused through the tail vein 1 h before streptozotocin administration. Beta-cell volume was measured by point counting of the insulin-containing cells. RESULTS: Zn2+ induced classical necrosis on MIN6N8 insulinoma cells which was associated with a rapid decline of intracellular ATP levels. Pyruvate inhibited Zn(2+)-induced necrosis of insulinoma cells and depletion of intracellular ATP by Zn2+. Pyruvate did not inhibit other types of necrosis or apoptosis. Energy substrates such as oxaloacetate, alpha ketoglutarate and succinic acid dimethylester also attenuated Zn(2+)-induced insulinoma cell death. Methylpyruvate that does not generate NAD+ in the cytoplasm or alpha-ketoisocaproate that stimulates ATP generation exclusively in mitochondria also protected insulinoma cells from Zn(2+)-induced necrosis. Pyruvate infusion inhibited the development of diabetes by protecting beta-cell mass after streptozotocin administration. CONCLUSION/INTERPRETATION: These results indicate that pyruvate inhibits Zn(2+)-induced necrosis of beta cells in vitro by protecting intracellular ATP levels and also streptozotocin-induced diabetes in vivo where Zn2+ has been reported to act as a paracrine death effector. PMID- 12898020 TI - Testing for linkage disequilibrium in the New Zealand radiata pine breeding population. AB - Linkage analysis is commonly used to find marker-trait associations within the full-sib families of forest tree and other species. Study of marker-trait associations at the population level is termed linkage-disequilibrium (LD) mapping. A female-tester design comprising 200 full-sib families generated by crossing 40 pollen parents with five female parents was used to assess the relationship between the marker-allele frequency classes obtained from parental genotypes at SSR marker loci and the full-sib family performance (average predicted breeding value of two parents) in radiata pine ( Pinus radiata D. Don). For alleles (at a marker locus) that showed significant association, the copy number of that allele in the parents was significantly correlated, either positively or negatively, with the full-sib family performance for various economic traits. Regression of parental breeding value on its genotype at marker loci revealed that most of the markers that showed significant association with full-sib family performance were not significantly associated with the parental breeding values. This suggests that over-representation of the female parents in our sample of 200 full-sib families could have biased the process of detecting marker-trait associations. The evidence for the existence of marker-trait LD in the population studied is rather weak and would require further testing. The exact test for genotypic disequilibrium between pairs of linked or unlinked marker loci revealed non-significant LD. Observed genotypic frequencies at several marker loci were significantly different from the expected Hardy-Weinberg equilibrium. The possibilities of utilising marker-trait associations for early selection, among-family selection and selecting parents for the next generation of breeding are also discussed. PMID- 12898021 TI - Mapping of quantitative trait loci associated with ultraviolet-B resistance in rice (Oryza sativa L.). AB - The detection of quantitative trait loci (QTLs) associated with UV-B resistance in rice should allow their practical application in breeding for such a complex trait, and may lead to the identification of gene characteristics and functions. Considerable variation in UV-B resistance exists within cultivated rice (Oryza sativa L.), but its detailed genetic control mechanism has not been well elucidated. We detected putative QTLs associated with the resistance to enhanced UV-B radiation in rice, using 98 BC(1)F(5) (backcross inbred lines; BILs) derived from a cross between Nipponbare (a resistant japonica rice variety) and Kasalath (a sensitive indica rice variety). We used 245 RFLP markers to construct a framework linkage map. BILs and both parents were grown under visible light with or without supplemental UV-B radiation in a growth chamber. In order to evaluate UV-B resistance, we used the relative fresh weight of aerial parts (RFW) and the relative chlorophyll content of leaf blades (RCC). The BIL population exhibited a wide range of variation in RFW and RCC. Using composite interval mapping with a LOD threshold of 2.9, three putative QTLs associated with both RFW and RCC were detected on chromosomes 1, 3 and 10. Nipponbare alleles at the QTLs on chromosome 1 and 10 increased the RFW and RCC, while the Kasalath allele at the QTL on chromosome 3 increased both traits. Furthermore, the existence of both QTLs on chromosomes 1 and 10 for UV-B resistance was confirmed using chromosome segment substitution lines. Plants with Kasalath alleles at the QTL on chromosome 10 were more sensitive to UV-B radiation than plants with them on chromosome 1. These results also provide the information not only for the improvement of UV-B resistance in rice though marker-associated selection, but also for the identification of UV-B resistance mechanisms by using near-isogenic lines. PMID- 12898022 TI - Chromosomal location of a race-specific resistance gene to Mycosphaerella graminicola in the spring wheat ST6. AB - Septoria tritici blotch, caused by Mycosphaerella graminicola, is a serious foliar disease of wheat worldwide. Qualitative, race-specific resistance sources have been identified and utilized for resistant cultivar development. However, septoria tritici blotch resistant varieties have succumbed to changes in virulence of M. graminicola on at least three continents. The use of resistance gene pyramids may slow or prevent the breakdown of resistance. A clear understanding of the genetics of resistance and the identification of linked PCR based markers will facilitate the recovery of wheat lines carrying multiple septoria tritici blotch resistance genes. The resistance gene in ST6 to isolate MG2 of M. graminicola was mapped with microsatellite markers in two populations, ST6/Erik and ST6/Katepwa. Bulk segregant analysis identified a marker on chromosome 4AL putatively linked to the resistance gene. A large linkage group was identified in each population using additional microsatellite markers mapping to chromosome 4AL. The resistance gene in ST6 mapped to the distal end of chromosome 4AL in each mapping population and was designated Stb7. Three of the microsatellite loci, Xwmc313, Xwmc219 and Xgwm160, mapped within 3.5 cM of Stb7; however, none flanked Stb7. Xwmc313 was the closest and mapped 0.3 and 0.5 cM from Stb7 in the crosses ST6/Katepwa and ST6/Erik, respectively. WMC313 will be very useful for marker-assisted selection of Stb7 in Canadian breeding programs because the ST6 allele of Xwmc313 was not identified in any of the Canadian common wheat cultivars tested. PMID- 12898023 TI - Mildew-resistant mutants induced in North American two- and six-rowed malting barley cultivars. AB - Mildew-resistant mutants were induced with sodium azide in three North American malting barley cultivars, two in the six-rowed Ursula (URS1 and URS2), one in the six-rowed Gertrud (GER1), and one in the two-rowed Prudentia (PRU1). Two of the mutants, URS1 and PRU1, showed complete resistance and were shown to have two new alleles at the mlo locus; these were designated, respectively, mlo31 and mlo32. Mutant URS2, showing partial resistance, was inherited as a dominant gene, but was not an allele at the Mla locus. The mean yield of each mutant was higher than that of its parental line, but yield levels varied across environments, although this was independent of the severity of the mildew attack. Other reasons, for example, the severity of the necrotic lesions in the mutants, may account for yield variations. The malting quality of the GER1 mutant proved similar to that of Gertrud, but both URS1 and URS2 showed lower malt extract than Ursula. This lower extract might be due to the smaller grain size of the mutants that could, in turn, result from necrotic lesions in the leaves, as implied by the effects on grain yield. PMID- 12898024 TI - Fine linkage mapping enables dissection of closely linked quantitative trait loci for seed dormancy and heading in rice. AB - Two quantitative trait loci (QTLs) for seed dormancy (tentatively designated Sdr1) and heading date ( Hd8) have been mapped to approximately the same region on chromosome 3 by interval mapping of backcross inbred lines derived from crosses between the rice cultivars Nipponbare (japonica) and Kasalath (indica). To clarify whether Sdr1 and Hd8 could be dissected genetically, we carried out fine-scale mapping with an advanced backcross progeny. We selected a BC(4)F(1) plant, in which a small chromosomal region including Sdr1 and Hd8, on the short arm of chromosome 3, remained heterozygous, whereas all the other chromosomal regions were homozygous for Nipponbare. Days-to-heading and seed germination rate in the BC(4)F(2) plants showed continuous variation. Ten BC(4)F(2) plants with recombination in the vicinity of Sdr1 and Hd8 were selected on the basis of the genotypes of the restriction fragment length polymorphism (RFLP) markers flanking both QTLs. Genotypes of those plants for Sdr1 and Hd8 were determined by advanced progeny testing of BC(4)F(4) families. Sdr1 was mapped between the RFLP markers R10942 and C2045, and co-segregated with C1488. Hd8 was also mapped between C12534S and R10942. Six recombination events were detected between Sdr1 and Hd8. These results clearly demonstrate that Sdr1 and Hd8 were tightly linked. Nearly isogenic lines for Sdr1 and Hd8 were selected by marker-assisted selection. PMID- 12898025 TI - Regional genetic differentiation in Western Australian sandalwood (Santalum spicatum) as revealed by nuclear RFLP analysis. AB - Western Australian sandalwood, Santalum spicatum, is widespread in the semi-arid and arid regions of Western Australia, and there is some morphological variation suggestive of two ecotypes. The level and structuring of genetic diversity within the species was investigated using anonymous nuclear RFLP loci. Santalum spicatum showed moderate levels of genetic diversity compared to other Australian tree species. The northern populations in the arid region showed greater levels of diversity and less population differentiation than the southern populations in the semi-arid region due to differences in the distribution of rare alleles. Equilibrium between drift and gene flow in the northern populations indicated that they have been established for a long period of time with stable conditions conducive to gene flow. In contrast, the southern populations showed a relationship between drift and gene flow indicative of a pattern of fragmentation and isolation where drift has greater effect than gene flow. The different patterns of diversity suggest that the ecotypes in the two regions have been subject to differences in the relative influences of drift and gene flow during their evolutionary history. PMID- 12898026 TI - Molecular studies on genetic integrity of open-pollinating species rye (Secale cereale L.) after long-term genebank maintenance. AB - The genetic integrity of six accessions represented by 14 sub-populations of the open-pollinating species rye ( Secale cereale L.) was investigated. Seeds available from a herbarium collection (first regeneration) and from the cold store (most recent regeneration) were multiplied two to fourteen times and fingerprinted using microsatellite markers. Four accessions had significantly different allele frequencies. These were multiplied seven to thirteen times. Nearly 50% of the alleles discovered in the original samples were not found in the material present in the cold store. However alleles were detected in the most recently propagated sub-populations, that were not observed in the investigated plants of the original one. The change in allele frequencies is a continuous process. Reasons for the occurrence of genetic changes and consequences for managing open pollinating species maintained in ex situ genebanks are discussed. PMID- 12898027 TI - Genetic and physical mapping of sequence-specific amplified polymorphic (SSAP) markers on the 1RS chromosome arm of rye in a wheat background. AB - Three rye-specific repeated sequences, pSc10C, pSc20H and R173-1, were used to design sequence-specific anchored primers. These primers and 16 restriction site specific adaptor primers were used in all possible combinations to establish sequence-specific amplified polymorphic (SSAP) markers for the 1RS chromosome arm of rye in a wheat background. Thirty 1RS-specific SSAP markers were detected in 19 primer combinations. Along with six markers localised previously on 1RS, 26 of the SSAP markers were mapped genetically in wheat genotypes carrying recombinant 1BL.1RS translocations. A clear decrease in recombination frequency from distal to proximal regions was observed. Wheat-rye addition lines for the 1R chromosome with different-sized deletions of the short arm were used to physically localise these markers. Physical mapping suggested an even distribution of the SSAP markers along the total length of the 1RS chromosome arm. PMID- 12898028 TI - AFLP analysis of genetic relationships among aromatic grapevines (Vitis vinifera). AB - Genotypic diversity has been detected among aromatic grapevines (Vitis vinifera) by molecular markers (AFLPs). The 22 primer-pairs generated a total of 1,331 bands of which 564 (40%) were polymorphic over all the genotypes. The bootstrap analysis pointed out that a large number of polymorphic bands (200-400) has to be used for a better estimation of the genetic distances among genotypes; 383 polymorphic AFLP bands were used for the cluster and the principal coordinate analyses because they did not present missing data across all the genotypes. The cluster analysis (UPGMA), based on polymorphic AFLP markers, revealed no relationship between the Moscato and Malvasia grapevines. The Malvasias, unlike the Moscatos distinguished by their distinct muscat aroma, have to be considered a more complex group because it includes muscat and non-muscat grapevines. The principal coordinate analysis (PCO) confirmed the pattern of the cluster analysis only for those varieties which presented a low coefficient of dissimilarity, while for the other varieties there was no correspondence between the two analyses. The pattern of aggregation among aromatic grapevines in the cluster and principal coordinate analyses does not support any classification that might include an aromatic grapevine group in V. vinifera. Even though some synonyms and homonyms are present among aromatic grapevines (V. vinifera), genetic diversity exists among genotypes in AFLP markers. PMID- 12898029 TI - Glycoalkaloids and acclimation capacity of hybrids between Solanum tuberosum and the incongruent hardy species Solanum commersonii. AB - F(1) and backcross hybrids between sexually incompatible species Solanum commersonii and Solanum tuberosum were characterized for glycoalkaloid content and capacity to cold acclimate. Glycoalkaloid (GA) analysis revealed that F(1) triploids and BC(1) pentaploids contained the glycoalkaloids of both parents. In BC(2) (near) tetraploids the situation was different, in that some hybrids produced the GAs of both parents, whereas others contained only the GAs of S. tuberosum. This suggested that the GAs from S. commersonii may be lost rapidly, and that they may have a simple genetic control. The total tuber GA content of BC(1) and BC(2) groups averaged quite acceptable levels (165.9 mg/kg in BC(1) and 192.8 mg/kg in BC(2)), with six genotypes having a GA content <200 mg/kg fresh weight. The F(1) triploid hybrids expressed a capacity to cold acclimate similar to S. commersonii, whereas BC(1) and BC(2) genotypes generally displayed an acclimation capacity higher than the sensitive parent but lower than S. commersonii. However, one BC(1) and two BC(2) genotypes with an acclimation capacity as high as S. commersonii were identified. The polar lipid fatty acid composition in S. commersonii and its hybrid derivatives showed that, following acclimation, there was a significant increase in 18:3. Correlation analysis between the capacity to cold acclimate and the increase in 18:3 was significant, suggesting that the increase in 18:3 can be used as a biochemical marker for the assisted selection of cold-acclimating genotypes in segregating populations. PMID- 12898030 TI - QTLs for agronomic traits in the Mediterranean environment identified in recombinant inbred lines of the cross 'Arta' x H. spontaneum 41-1. AB - A genetic linkage map has been developed for recombinant inbred lines (RILs) of the cross 'Arta' x Hordeum spontaneum 41-1. One hundred and ninety four RILs, randomly chosen from a population of 494 RILs, were mapped with 189 markers including one morphological trait ( btr = brittle rachis locus). The linkage map extended to 890 cM. Agronomic traits such as grain yield, biological yield, days to heading, plant height, cold tolerance and others were evaluated at the ICARDA research stations Tel Hadya and Breda during the years 1996-97 and 1997-98. QTLs for agronomic traits related to drought resistance were localized. For the most important character 'plant height under drought stress', QTLs on 2H, 3H and 7H were detected. The 'plant height' QTLs, specially the one on 3H, showed pleiotropic effects on traits such as days to heading, grain yield and biological yield. QTLs were also identified for other traits associated with adaptation to the Mediterranean environment such as cold tolerance, days to heading and tiller number. The identification of QTLs for agronomic traits is a first step to analyze and to dissect complex characters such as adaptation to drought tolerance. PMID- 12898031 TI - An integrative genetic linkage map of winter wheat (Triticum aestivum L.). AB - We constructed a genetic linkage map based on a cross between two Swiss winter wheat ( Triticum aestivum L.) varieties, Arina and Forno. Two-hundred and forty F(5) single-seed descent (SSD)-derived lines were analysed with 112 restriction fragment length polymorphism (RFLP) anonymous probes, 18 wheat cDNA clones coding for putative stress or defence-related proteins and 179 simple-sequence repeat (SSR) primer-pairs. The 309 markers revealed 396 segregating loci. Linkage analysis defined 27 linkage groups that could all be assigned to chromosomes or chromosome arms. The resulting genetic map comprises 380 loci and spans 3,086 cM with 1,131 cM for the A genome, 920 cM for the B genome and 1,036 cM for the D genome. Seventeen percent of the loci showed a significant ( P < 0.05) deviation from a 1:1 ratio, most of them in favour of the Arina alleles. This map enabled the mapping of QTLs for resistance against several fungal diseases such as Stagonospora glume blotch, leaf rust and Fusarium head blight. It will also be very useful for wheat genetic mapping, as it combines RFLP and SSR markers that were previously located on separate maps. PMID- 12898032 TI - [Hematuria in cases of bladder cancer]. AB - In the majority of cases, painless macroscopic hematuria is a typical initial symptom of bladder carcinoma. The time of occurrence (early and late symptom) neither correlates with tumor size and degree nor with infiltration depth. The respective testing method and threshold value exert a significant influence on frequency and constancy of a potential microscopic hematuria. Without sufficient standardization, however, it is understandable that even the existing guidelines for diagnosis of hematuria recommend different methods and threshold values. This paper provides an overview of the various testing methods and threshold values in microscopic hematuria, their influence on the diagnosis of bladder carcinoma as well as the possibility of differentiating the source of hematuria morphologically. PMID- 12898033 TI - [Urinary cytology in cases of bladder cancer: a critical evaluation]. AB - Is urinary cytology still a useful method in cases of bladder cancer since additional methods (fluorescence image analysis, monoclonal antibody immunocytology, fibrin degradation products, nuclear matrix protein measurement and others) are under investigation? The literature has been reviewed and compared with classical and new cytology statistics: urinary cytology is less useful than cystoscopy and some of the urine bound tests for the diagnosis of low grade tumors. On the other hand, cytology is extremely valuable for the diagnosis of high grade transitional cell cancers (TCC) and especially carcinomas in situ. The presence of high-grade TCC in the cytology specimen from a patient with low grade papillary TCC suggests either an unrecognised carcinoma in situ or high grade disease in the upper urinary tract or urethra. Urinary cytology is still indispensable in the management of patients with transitional cancer. PMID- 12898035 TI - [Polymerase chain reaction in the urinary diagnosis of bladder cancer]. AB - The polymerase chain reaction (PCR) is a highly sensitive and specific method for the detection of genetic material. Over the last few years, this method has been used increasingly for the molecular detection of disease. This report demonstrates the use of PCR in the diagnosis of bladder cancer. The principles of the method are shown by describing the detection of p53, CD44 and telomerase, as well as in microsatellite analysis. PMID- 12898034 TI - [Non-invasive urinary diagnosis of bladder cancer. What do we know?]. AB - Although the current system of classifying bladder cancer by stage and histological grade is very useful, it is still difficult to predict the natural progression of the disease either with or without therapy. Cystoscopy and urine cytology are currently the gold standards in the monitoring and diagnosis of bladder cancer. Classical urine cytology is, however, at least in the diagnosis of G1-tumors, characterized by a relatively low sensitivity. In the last few years, the molecular biological investigation of the basic mechanisms involved in carcinogenesis has provided a host of markers which are of potential diagnostic value for bladder cancer. We provide a current, comprehensive review of the literature on bladder tumor markers and summarize their diagnostic and prognostic potential. At present, no diagnostic marker with a comparable sensitivity and specificity to cystoscopy exists, given that cystoscopy has never been evaluated. The combined analysis of several tumor markers seems to be the most promising approach as an adjunct to cystoscopy. Moreover, the increasing simplification of test systems will increase their acceptance by clinicians. PMID- 12898036 TI - [Biochips in bladder cancer research]. AB - The techniques of DNA microarrays, protein arrays, and tissue microarrays complement one another and in the future the combination of these methods will possibly enable the detection of genes specific for bladder cancer. The SELDI technology (surface-enhanced laser desorption/ionization) will then be used to prove if the genes are translated into proteins and if those proteins have a relevant effect on the development and progression of the cancer disease. The consequence is the facilitation of the development of prognostic markers for this disease entity, which can then be easily tested in a large study population by tissue microarray. Beyond the gain of new insights into the biological understanding of the development and progression of bladder cancer, these techniques will hopefully enable the design of new therapeutic concepts, which apply to the individual biology of the tumor. PMID- 12898037 TI - [Vasectomy today: a review of 1,275 vasectomies in 10 years]. AB - We report on a large series of 1,275 patients who underwent outpatient vasectomy performed by a single urological surgeon within a 10-year period. In addition, the results of a prospective questionnaire-based study on 217 patients regarding their opinions and motivation, and the financial aspects of the vasectomy are discussed. The mean age was 37.0+/-5.9 years. A total of 98.4% of the patients had an uneventful postoperative course. The average duration of sick leave was 2.2+/-0.21 days. During the study period, the length of sick leave dropped significantly from 2.63 (1990/91) to 1.1 days (1998/99; P=0.001). Efficacy was documented with azoospermia rates of 96.6% and 98.5% in patients who presented for two and three sperm examinations, respectively. A vast majority of patients (94.6%) felt that the procedure should be covered by their health insurance, although 88.1% stated they would also pay by themselves. A minority of patients (0.7%: 9/1,275) subsequently considered vasectomy reversal. The wish to reverse the vasectomy was significantly associated with a younger patient age. In conclusion, outpatient vasectomy provides a safe and reliable form of contraception at low cost. Overall satisfaction in appropriately counselled patients is very high. Based on these findings, further attempts to propagate vasectomy as a timely form of contraception are medically and socioeconomically recommended. PMID- 12898038 TI - [Prostate cancer diagnosis using ultrasound elastography. Introduction of a novel technique and first clinical results]. AB - During the last decade screening has improved prostate cancer detection. The main reason for this development is a better understanding of the margins of prostate specific antigen (PSA) serum levels and the classification of PSA subtypes. In contrast, the introduction of transrectal ultrasound has not led to a measurable change in the prostate cancer detection rate. Our aim was to develop a novel ultrasound system for the acquisition of elastographic images of the prostate and evaluate the system regarding its clinical applicability. We used a technically modified conventional ultrasound system and analyzed the high-frequency ultrasonic data with a computer program. The first patient-based results suggest that elastography allows an accurate measurement of tumor size and localization in contrast to conventional transrectal ultrasound. Elastography visualizes different tissue elasticities to distinguish benign and cancerous tissue. Thus, we were able to even correctly classify prostate cancer lesions which are iso- or hyperechoic in B-mode sonography. PMID- 12898039 TI - [Familial versus sporadic prostate cancer in the German population. Clinical and pathological characteristics in patients after radical prostatectomy]. AB - Family history is one of the strongest epidemiological risk factors for the development of prostate cancer. The impact on the clinical presentation and prognosis, however, is controversial. In the present study, we analyzed 464 familial and 492 sporadic prostate cancer patients following radical prostatectomy. The average age at onset was 62.1 years in the familial group and 64.2 years in the sporadic controls (p<0.001). The screening attitude, DRE findings and the PSA values at diagnosis the pT- and pN-stages, and the tumor grade did not differ between both groups. With a median follow-up of 3.3 years, the 5- and 10-year progression-free survival rates were 76.2% and 56.5% in familial and 70.8% and 55.5% in sporadic patients, respectively (n.s.). A multiple logistic regression analysis revealed that family history did not have an influence on disease recurrence. In our population there was no association between a familial predisposition and clinical features or clinical course of the disease. Whether hereditary prostate cancer is distinct from sporadic forms cannot be determined before the underlying genetic alterations are identified. PMID- 12898040 TI - [Ureterocystoplasty: functional results and possible problem areas]. AB - Surgical procedures such as ileal augmentation cystoplasties are often necessary in neurogenic low-compliance bladders. Whenever possible one should avoid using segments of the GI tract in the urinary tract and preferably use the patient's own ureter for augmentation cystoplasty. This procedure includes transperitoneal nephrectomy of a nonfunctioning kidney with preservation of the renal pelvis and the megaureter, followed by detubularization of the ureter and its integration into the opened urinary bladder. We performed this procedure on four patients, two boys and two girls (aged 6-13 years). In the long-term follow-up they all had good rehabilitation results for their lower urinary tracts with an increase of the storage volume and normalization of their reduced compliance. No deterioration of the remaining upper urinary tract was found. Complications such as partial ureteral necrosis have to be taken into consideration when the vascularization of the ureter is compromised, for example, by multiple prior antireflux procedures. PMID- 12898042 TI - [Psychopharmacological treatment in the pre-clinical emergency medicine]. AB - Psychiatric emergencies are a common cause for a call for the pre-clinical emergency physician (EP). However, previous studies reported serious problems in diagnosing and particularly treating these patients. Although evidence-based treatment guidelines and algorithms exist for the most important somatic disorders the EP has to deal with, they do not for psychiatric emergencies. It is the aim of this review, subsequent to an extensive overview on the available literature, to provide the reader with differentiated suggestions for pharmacological crisis intervention as well as with treatment recommendations for different psychiatric disorders with an emphasis on the special needs in the pre clinical emergency medicine. After conducting a literature research from 1971 to 2002, 31 double-blind trials were found comparing different antipsychotics and benzodiazepines for efficacy and tolerability. Further, pharmacological data and product information of the most commonly used drugs were evaluated. Following, antipsychotics and benzodiazepines were generally assessed for their usefulness in the pre-clinical emergency medicine, the advantages of different drugs for specific indications are presented and dosage recommendations are given. As an antipsychotic, haloperidol still is the drug of choice. Within the benzodiazepines, lorazepam seems to have advantages over diazepam. An antipsychotic medication is particularly indicated for the treatment of schizophrenia, mania, drug-induced psychoses and delirium whereas benzodiazepines are favourable for the treatment of anxiety, restlessness and agitation due to neurotic or reactive circumstances. There is some evidence suggesting that newer generation, so-called "atypical" antipsychotics may play a role in the treatment of psychiatric emergencies, however, controlled trials are necessary to substantiate their potential benefits in the preclinical emergency medicine. PMID- 12898041 TI - Not just for housekeeping: protein initiation and elongation factors in cell growth and tumorigenesis. AB - Proteins provide the structural framework of a cell and perform the enzymatic activities sustaining DNA replication and energy production. The hormones and growth factors that facilitate organ-to-organ communication are proteins as are the receptors and signaling intermediaries that integrate extracellular stimuli to intracellular action. As such, eukaryotic cells devote tremendous effort and energy to protein synthesis. The enzymes involved in protein synthesis have traditionally been described as cellular housekeepers. This was meant to imply that while they were necessary for cell viability, they were not thought to have a causal role in activating cell differentiation or neoplastic development the way that a transcription factor or hormone receptor might. However, two protein translation factors, protein initiation factor eIF4E and protein elongation factor eEF1A2, have been identified as important human oncogenes. This review summarizes recent work showing that protein initiation and elongation factors have important regulatory roles in cell growth, apoptosis, and tumorigenesis. PMID- 12898043 TI - [Prophylaxis of postoperative nausea and vomiting (PONV) in children undergoing strabismus surgery. Sevoflurane/N2O plus dimenhydrinate vs.propofol/remifentanil plus dimenhydrinate]. AB - BACKGROUND AND STUDY GOALS: Strabismus surgery in children is frequently associated with a very high incidence of postoperative nausea and vomiting (PONV). In this study we investigated, whether TIVA is superior to inhalation anaesthesia concerning PONV when a prophylaxis using dimenhydrinate (diphenhydramine chlorotheophyllate) is administered in both groups. METHODS: 110 patients (3-10 years) were prospectively and randomly allocated to one of the following groups: Group TD (TIVA/dimenhydrinate; n= 55): anaesthesia was induced and maintained using remifentanil and propofol in common clinical dosages. Patients were ventilated with 30% O(2) in air. Group VD (Volatile/ dimenhydrinate; n= 55): anaesthesia was induced and maintained with sevoflurane in common clinical concentrations together with N(2)O in 30% O(2). All patients received dimenhydrinate 1 mg.kg(-1) i.v. after induction. PONV was recorded within the first 24 h postoperatively. The chi-square test and a multivariate analysis were used for statistical analysis. RESULTS: 52 patients of group TD and 54 of group VD were analysed. There was a trend in the incidence of postoperative nausea (PN): group TD 17%, 95% CI: 8-30%) compared to group VD 31%, 95% CI: 20 46%), p = 0.09. No difference was seen for PV and PONV: 21% (95% CI: 11-35%) in group TD vs. 35% (95% CI: 23-49%) in group VD, p = 0.109. These non-significant results can be easily attributed to the lack of power about 30%) of this study. According to the results of the multivariate analysis volatile anaesthesia was identified as an independent risk factor for PONV (OR: 2.92, 95% CI: 1.02-8.36). Other variables that were found to be an independent risk factor included history of PONV (OR: 8.19, 95% CI: 1.84-36.43), surgery lasting longer than 30 min (OR: 5.89, 95% CI: 1.82-19.82) and "Faden-operations" (retroequatorial myopexy) (OR: 5.48, 95% CI: 1.74-17.21). CONCLUSIONS: TD only showed a trend to lower PN incidences and no differences as for PV and PONV incidences, most likely due to a lack of power of this study. However, according to the results of the multivariate analysis, inhalation anaesthesia was shown to be an independent risk factor for PONV, as were history of PONV, surgery >30 min and "Faden-operations". PMID- 12898044 TI - [Cognitive functions and cerebral oxygenation of older patients after general and regional anaesthesia]. AB - OBJECTIVE: The aim of the present study was to show the influence of cerebral oxygenation (regional cerebral oxygen saturation rSO(2) by near infrared spectroscopy) and of the nocturnal arterial oxygen saturation (SatO(2) by pulse oximetry) on the restitution of cognitive functions in patients aged between 40 and 85 years scheduled for elective hip arthroplasty. METHODS: A total of 40 patients (ASA II) were randomized to be operated either in general anaesthesia or regional anaesthesia. The patients were additionally classified by age (40-64 years and 65-85 years). Cognitive functions were tested 14-16 h pre-operatively (t0), 1.5 h post-operatively (t1) and at the first and third postoperative days (t2 and t3). During testing, as well as during surgery and postoperatively until t1, rSO(2) was continuously measured. SatO(2) was measured in the night before surgery (N0) and for 3 nights after surgery (N1, N2, N3) between 22.00 p.m. and 5.00 a.m. These measurements were divided into interval groups (80-83%, 84-87%, 88-91%, 92-95%, 96-100%). RESULTS: Almost all cognitive functions were significantly reduced at t1 in all groups compared to t0, but recovered up to the third postoperative day (t3). RSO(2) in contrast was significantly reduced in all groups compared to t0 at the third postoperative day (t3). The relative proportion of the intervals compared to the total measurement time for SatO(2) shifted in both anaesthesia procedures: before surgery (N0) the most frequented interval was 96-100%, after surgery (N1, N2) it was 88-91%. There was no correlation between rSO(2), the restitution of the tested cognitive functions and SatO(2). CONCLUSIONS: Cognitive functions recovered completely during the first 3 postoperative days in patients scheduled for elective hip surgery under general or regional anaesthesia regardless of age and type of anaesthesia. This restitution of cognition occurred despite a significant decrease of cerebral oxygenation (rSO(2)) and despite an increase of nocturnal hypoxaemic intervals. Changes of the rsO(2) up to 3% below the baseline values (measured by NIRS) do not predict cognitive restitution. A minimal limiting value of the rSO(2) could not be defined. PMID- 12898045 TI - [Knotting of a catheter in the plexus brachialis. A rare complication]. AB - For the repair of an open injury to the hand, a continuous axillary block was performed in a 40- year-old male patient. Slight resistance was experienced during advancement of the catheter. Adequate surgical anaesthesia developed after the injection of 40 ml of a bupivacaine/prilocaine mixture. For postoperative analgesia an infusion of bupivacaine 0.25% was started. Three days later the removal of the catheter proved to be difficult due to a knot in the distal part of the catheter. This seems to be the first report of a knot in a catheter used for continuous axillary plexus block. PMID- 12898046 TI - [Central anticholinergic syndrome and apnea after general anaesthesia. A rare manifestation of the central anticholinergic syndrome]. AB - The central anticholinergic syndrome (CAS) is a rarely observed condition after general anaesthesia. There are no definitive criteria to set the diagnosis of CAS. The syndrome may manifest in clinical neurological signs, such as hyperactive states or a depressed CNS state. The diagnosis usually depends on a process of exclusion of other conditions and is confirmed after rapid recovery following administration of physostigmine. We report on a 34-year-old patient who suddenly lost consciousness and developed respiratory arrest 1 h after general anesthesia and normal postoperative recovery. CAS was considered, although apnea has not been reported as a clinical symptom of this disease up to now and no peripheral signs of CAS were observed. After the administration of 1 mg physostigmine the symptoms resolved immediately and the patient started sufficient spontaneous breathing. However, corresponding to the plasma elimination half-life of the drug, further comatose episodes with apnea occurred. Therefore, the patient was admitted to the ICU and an infusion of physostigmin at a rate of up to 5 mg/h was started. Due to this therapy the patient's state became stable and 15 h after the first manifestation of CAS the infusion of physostigmin was discontinued. The following postoperative course was uneventful. In case of reduced vigilance with apnea after general anaesthesia, central anticholinergic syndrome should be considered. For diagnostic and therapeutic purposes the administration of physostigmine should be attempted. PMID- 12898047 TI - [Current treatment of chronic heart failure]. AB - Cardiovascular diseases including heart failure represent a common disease in patients referred for anesthesia. In most cases, heart failure is caused by left ventricular dysfunction due to coronary heart disease. The aims of the treatment of chronic heart failure are the relief of symptoms, the improvement of prognosis and the prevention of the progression of heart failure. The first-line treatment involves the underlying heart disease such as myocardial revascularisation procedures in coronary heart disease or the correction of valve diseases. The pharmacological therapy depends on the stage of heart failure and symptoms of the patient. Heart failure therapy includes ACE-inhibitors, betablockers, diuretics und digitalis. Nitrates can be prescribed in patients with symptomatic heart failure despite adequate therapy but calcium antagonists are not recommended. Repeated or prolonged treatment with positive inotropic agents like phosphodiesterase inhibitors or beta-adrenergic drugs increases mortality but this is commonly used in acute stages of heart failure refractory to treatment. Interactions of ACE-inhibitors or AT1- antagonists with anesthetic agents can lead to severe hypotension especially in hypovolemic patients. Whether those drugs should be continued perioperatively or not has been controversially discussed. The use of betablockers has a positive impact on cardiac morbidity and mortality during and early after surgery. Chronic treatment with diuretics can be associated with hypovolemia and an imbalance of electrolytes leading to hypotension and arrhythmia during anesthesia but careful evaluation prior to anesthesia can avoid such complications. The continuation of digitalis during anesthesia has been controversially discussed due to the various interactions with anesthetics. PMID- 12898049 TI - [Informing non-German speaking patients about medical problems: three new decisions on an old theme]. PMID- 12898048 TI - [Vertical infraclavicular blockade of the brachial plexus (VIP). A modified method to verify the puncture point under consideration of the risk of pneumothorax]. AB - INTRODUCTION: The vertical infraclavicular blockade of the brachial plexus (VIP) according to Kilka et al.is a technique which has gained more importance over the past years. This method distinguishes itself from other periclavicular techniques by a very low risk of pneumothorax (0.2%), which seems to be increased with asthenic patients. METHODS: In the study presented we examined 52 patients undergoing a vertical infraclavicular blockade of the brachial plexus, for an alternative method to determine the puncture point. With 31 of the 52 patients, who had a small distance (<20 cm) between the landmarks jugulum and anterior process of the acromion, the puncture point was moved 0.3 cm in a lateral direction for each centimeter less than 20 cm. Additionally we determined the "finger-point", i.e. the medial margin of the anesthetist's index finger, placed in the gap between the M. deltoideus and M. pectoralis with the finger tip touching the clavicle. RESULTS: In 54% of the patients, the "finger-point" corresponded to the measured puncture point. In 46% of the patients, these points varied by a maximum of 1 cm in the lateral or medial direction. In 53% of the patients, the plexus could be found at the measured puncture point,which applied especially to the patients with a small distance between the leading points (<20 cm) and as a consequence a lateralized puncture point. If a correction of the puncture point was necessary to find the plexus, the correction by skin movement would always be in the direction of the "finger-point". CONCLUSIONS: As a consequence, we assume that if the distance between the leading points jugulum and ventral process of acromion is smaller than 20 cm, the puncture point for a vertical infraclavicular blockade of the brachial plexus should be lateralized as described above; additionally, the "finger-point" should be determined in order to verify the puncture point and to finally give an idea of the direction, in case of a possible need for correcting the puncture point. PMID- 12898050 TI - [Congress on interdisciplinary intensive medicine: 9th to 14th March 2003, Garmisch-Partenkirchen]. PMID- 12898051 TI - [Anaphylactic effects after muscle relaxants. Exchange of opinions of the paper "Anesthesia in allergic diathesis" by H. Kisch-Wedel and M. Thiel. In : Der Anaesthesist (2002) 51:868-881]. PMID- 12898052 TI - [Allergic diathesis]. PMID- 12898055 TI - [Brachial plexus. Anesthesia and analgesia]. AB - This review explains the different approaches to the brachial plexus (posterior cervical, interscalene, supra- and infraclavicular, and axillary) and their advantages and disadvantages (indications, contraindications, and complications) for surgery and postoperative or chronic pain management. One of the focussed areas of this review is the use of continuous catheter techniques. Information about the most commonly used local anesthetics as well as adjuncts suggested in the literature is summarized. As essential components for the success of those techniques, organizational and documentation requirements are described. In summary, regional techniques for single shot or continuous block of the brachial plexus are an efficient and safe way of providing anesthesia and analgesia for surgery or pain in the region of the shoulder, arm, or hand. PMID- 12898056 TI - Fibrous dysplasia complicated by aneurysmal bone cyst formation affecting multiple cervical vertebrae. AB - Fibrous dysplasia is a well-known benign dysplastic process of bone. However, fibrous dysplasia is very uncommon in the spine. Further, to our knowledge, coexistence of fibrous dysplasia and aneurysmal bone cyst in the spine has not been reported. This manuscript presents a patient who had both processes involving the cervical spine. PMID- 12898057 TI - Percutaneous drainage of a postoperative intraspinal hematoma using a Tuohy needle. AB - A 78-year-old man developed a large subdural hematoma extending from T12 to L3 levels following L3 laminectomy and L3-5 posterior spinal fusion. He had associated neurological signs and symptoms. MR imaging showed typical signal characteristics of a subacute intraspinal subdural hematoma. Percutaneous drainage was successfully performed under CT guidance by inserting a Tuohy needle through the L3 laminectomy defect. The catheter packaged with the Tuohy needle was inserted cranially into the hematoma and 30 ml of blood was aspirated. Follow up MR imaging confirmed resolution of the hematoma and the patient made a rapid recovery. PMID- 12898059 TI - The migration of femoral components after total hip replacement surgery: accuracy and precision of software-aided measurements. AB - OBJECTIVE: To assess the accuracy and precision of a software-aided system to measure migration of femoral components after total hip replacement (THR) on digitised radiographs. DESIGN AND PATIENTS: Subsidence and varus-valgus tilt of THR stems within the femur were measured on digitised anteroposterior pelvic radiographs. The measuring software (UMA, GEMED, Germany) relies on bony landmarks and comparability parameters of two consecutive radiographs. Its accuracy and precision were calculated by comparing it with the gold standard in migration measurements, radiostereometric analysis (RSA). Radiographs and corresponding RSA measurements were performed in 60 patients (38-69 years) following cementless THR surgery. RESULTS AND CONCLUSIONS: The UMA software measured the subsidence of the stems with an accuracy of +/-2.5 mm and varus valgus tilt with an accuracy of +/-1.8 degrees (95% confidence interval). A good interobserver and intraobserver reliability was calculated with Cronbach's alpha ranging from 0.86 to 0.97. Measuring the subsidence of THR stems within the femur is an important parameter in the diagnosis of implant loosening. Software systems such as UMA improve the accuracy of migration measurements and are easy to use on routinely performed radiographs of operated hip joints. PMID- 12898058 TI - MRI demonstration of intra-articular median nerve entrapment after elbow dislocation. AB - We report a case of median nerve entrapment in the elbow joint diagnosed with MRI 14 months after closed reduction of a dislocated elbow joint. PMID- 12898060 TI - Polychlorinated biphenyl (PCB) degradation and persistence of a gfp-marked Ralstonia eutropha H850 in PCB-contaminated soil. AB - Ralstonia eutropha H850 was labelled chromosomally with a gfp marker gene encoding for the green fluorescent protein, and designated R. eutropha H850g13. Visual observation of green fluorescent cells under an epifluorescence microscope, and PCR amplification products, confirmed that the bacterium was labelled with gfp. Southern blot hybridization products further confirmed the gfp was chromosomally labelled. Using resting cell assays, it was determined that insertion of the gfp gene decreased the microorganisms' ability to degrade biphenyl compared to the parent strain. However, this marker facilitated the identification and monitoring of R. eutropha H850g13 survival in soil microcosm experiments. Survival and polychlorinated biphenyl degradation by R. eutropha H850g13 was analysed in soil microcosms spiked with 2,2',5,5'-tetrachlorobiphenyl (TeCB). R. eutropha H850g13 was detected by viable plate counts and most-probable number/PCR after 102 days in TeCB-contaminated soil microcosms, and was likely outcompeted by indigenous soil microorganisms in microcosms amended with oil and Daramend (an organic amendment, http://www.adventusremediation.com). R. eutropha H850g13 did not degrade TeCB in any of the soil microcosms. This research confirmed that gfp was useful as a marker to distinguish R. eutropha H850g13 from indigenous soil microorganisms over a 102 day period and that, under the experimental conditions used, R. eutropha H850g13 did not degrade TeCB. PMID- 12898061 TI - Azotobacter vinelandii mutants that overproduce poly-beta-hydroxybutyrate or alginate. AB - Azotobacter vinelandii produces two polymers of industrial importance, i.e. alginate and poly-beta-hydroxybutyrate (PHB). Alginate synthesis constitutes a waste of substrate when seeking to optimize PHB production and, conversely, synthesis of PHB is undesirable when optimizing alginate production. In this study we evaluated the effect of a mutation in algA, the gene encoding the enzyme that catalyzes the first step of the alginate biosynthetic pathway in the production of PHB. We also evaluated production of alginate in strain AT6 carrying a phbB mutation that impairs PHB synthesis. The algA mutation prevented alginate production and increased PHB accumulation up to 5-fold, determined in milligrams per milligram of protein. Similarly, the phbB mutation increased alginate production up to 4-fold. PMID- 12898062 TI - Molecular cloning and characterization of a laccase gene from the basidiomycete Fome lignosus and expression in Pichia pastoris. AB - A cDNA encoding for a laccase was isolated from the white-rot fungus Fome lignosus by RT-PCR. It contained an open reading frame of 1,557 bp. The deduced mature protein consisted of 497 amino acids and was preceded by a signal peptide of 21 amino acids. The genomic DNA of the laccase, containing 11 introns, was cloned by PCR. The cDNA was cloned into the vectors pGAPZalphaA and pGAPZA, and expressed in the Pichia pastoris GS115. Laccase-secreting transformants were selected by their ability to oxidize the substrate 2'2-azinobis-(3 ethylbenzthiaoline-6-sufonic acid) (ABTS). The laccase activity obtained with the native signal peptide was found to be fivefold higher than that obtained with the alpha-factor secretion signal peptide. The presence of 0.4 mM copper was necessary for optimal activity of the enzyme. The highest activity value reached 9.03 U ml(-1), and the optimal secreting time was 2~3 days at 20 degrees C. The crude laccase was stable in a pH range from 6.0 to 10.0 and at temperatures lower than 30 degrees C in pH 4.5 for 24 h. The molecular mass of the enzyme was estimated to be 66.5 kDa by SDS-PAGE. The optimum pH and temperature were 2.4 and 55 degrees C. The Km and Vmax values for ABTS were 177 microM and 23.54 micromol min(-1) respectively. The extent of glycosylation of the purified enzyme was 58.6%. PMID- 12898063 TI - Bioadsorption of cadmium ion by cell surface-engineered yeasts displaying metallothionein and hexa-His. AB - The Cd(2+)-chelating abilities of yeast metallothionein (YMT) and hexa-His displayed on the yeast-cell surface were compared. Display of YMT and hexa-His by alpha-agglutinin-based cell-surface engineering was confirmed by immunofluorescent labeling. Surface-engineered yeast cells with YMT and hexa-His fused in tandem showed superior cell-surface adsorption and recovery of Cd2+ under EDTA treatment on the cell surface than hexa-His-displaying cells. YMT was demonstrated to be more effective than hexa-His for the adsorption of Cd2+. Yeast cells displaying YMT and/or hexa-His exhibited a higher potential for the adsorption of Cd2+ than Escherichia coli cells displaying these molecules. In order to investigate the effect of the displayed YMT and hexa-His on sensitivity to toxic Cd2+, growth in Cd(2+)-containing liquid medium was monitored. Unlike hexa-His-displaying cells, cells displaying YMT and hexa-His fused in tandem induced resistance to Cd2+ through active and enhanced adsorption of toxic Cd2+. These results indicate that YMT-displaying yeast cells are a unique bioadsorbent with a functional chelating ability superior to that of E. coli. PMID- 12898064 TI - Inhibiting mild steel corrosion from sulfate-reducing bacteria using antimicrobial-producing biofilms in Three-Mile-Island process water. AB - Biofilms were used to produce gramicidin S (a cyclic decapeptide) to inhibit corrosion-causing, sulfate-reducing bacteria (SRB). In laboratory studies these biofilms protected mild steel 1010 continuously from corrosion in the aggressive, cooling service water of the AmerGen Three-Mile-Island (TMI) nuclear plant, which was augmented with reference SRB. The growth of both reference SRB (Gram-positive Desulfosporosinus orientis and Gram-negative Desulfovibrio vulgaris) was shown to be inhibited by supernatants of the gramicidin-S-producing bacteria as well as by purified gramicidin S. Electrochemical impedance spectroscopy and mass loss measurements showed that the protective biofilms decreased the corrosion rate of mild steel by 2- to 10-fold when challenged with the natural SRB of the TMI process water supplemented with D. orientis or D. vulgaris. The relative corrosion inhibition efficiency was 50-90% in continuous reactors, compared to a biofilm control which did not produce the antimicrobial gramicidin S. Scanning electron microscope and reactor images also revealed that SRB attack was thwarted by protective biofilms that secrete gramicidin S. A consortium of beneficial bacteria (GGPST consortium, producing gramicidin S and other antimicrobials) also protected the mild steel. PMID- 12898065 TI - Overexpression of the ATP-dependent helicase RecG improves resistance to weak organic acids in Escherichia coli. AB - Increased resistance to several weak organic acids was conferred on Escherichia coli by overexpression of the ATP-dependent helicase RecG and, to a lesser extent, by overexpressing the helicase RuvAB. This property of helicases was identified by reproducible selection of recG-bearing clones from genomic libraries of the acetate-resistant species Acetobacter aceti and Staphylococcus capitis. We show that overexpression of RecG from both species, but also from E. coli, increased the maximum biomass concentration attained by E. coli cultures that were grown in the presence of various weak organic acids and uncouplers. Furthermore, overexpression of RecG from A. aceti significantly improved the maximum growth rates of E. coli under weak organic acid challenge. Based on the known role of RecG in DNA replication/repair, our data provide a first indication that weak organic acids negatively affect DNA replication and/or repair, and that these negative effects may be counteracted by helicase activity. PMID- 12898066 TI - Growth characteristics and oxidative capacity of Acetobacter aceti IFO 3281: implications for L-ribulose production. AB - We studied the growth characteristics and oxidative capacities of Acetobacter aceti IFO 3281 in batch and chemostat cultures. In batch culture, glycerol was the best growth substrate and growth on ethanol occurred only after 6 days delay, although ethanol was rapidly oxidized to acetic acid. In continuous culture, both glycerol and ethanol were good growth substrates with similar characteristics. Resting cells in a bioreactor oxidized ribitol to L-ribulose with a maximal specific rate of 1.2 g g(-1) h(-1)). The oxidation of ribitol was inhibited by ethanol but not by glycerol. Biomass yield (Y(SX); C-mmol/C-mmol) on ethanol and glycerol was low (0.21 and 0.17, respectively). In the presence of ribitol the yield was somewhat higher (0.25) with ethanol but lower (0.13) with glycerol, with respectively lower and higher CO(2) production. In chemostat cultures the oxidation rate of ribitol was unaffected by ethanol or glycerol. Cell-free extract oxidized ethanol very slowly but not ribitol; the oxidative activity was located in the cell membrane fraction. Enzymatic activities of some key metabolic enzymes were determined from steady-state chemostat with ethanol, glycerol, or ethanol/glycerol mixture as a growth limiting substrate. Based on the measured enzyme activities, metabolic pathways are proposed for ethanol and glycerol metabolism. PMID- 12898067 TI - A molecule bearing an immunoglobulin-like V region of the CTX subfamily in amphioxus. AB - An expressed sequence tag with significant similarity to a vertebrate T-cell receptor (Tcr) sequence was found in a cDNA library prepared from the posterior part of the adult amphioxus, Branchiostoma lanceolatum ( Brla). Characterization of the corresponding cDNA clone revealed the presence of an open reading frame encoding a 351 amino acid residue-long polypeptide. The putative protein, tentatively designated Brla-VDB for "V-domain bearing", appears to consist of two domains. The N-terminal domain begins with a putative leader peptide followed by a sequence resembling the V domain of the CTX protein originally found to be expressed in the cortical thymocytes of the clawed frog Xenopus. The C-terminal domain of the VDB protein does not show significant sequence similarity to any entry in the databases and contains five hydrophobic segments separated by short intervening hydrophilic stretches. It may therefore belong to a protein that crosses the plasma membrane five times. These findings support the notion that V domains resembling those found in Tcrs evolved in nonvertebrates before the emergence of the adaptive immune system and may have participated in functions not involved directly in immunity. PMID- 12898069 TI - Ultrasonographic findings after laparoscopic repair of paediatric female inguinal hernias: the 'vanishing rosebud'. AB - BACKGROUND: Laparoscopic surgery is a good alternative to the open technique for treating hernias in female paediatric patients. The laparoscopically inverted and sutured hernia sac forms a nodule, the long-term fate of which has not been previously studied radiologically. OBJECTIVE: To describe the early and delayed US changes after laparoscopic inversion and suturing of paediatric female inguinal hernias. MATERIALS AND METHODS: Twenty girls (age 1.5 months to 12 years; median 4.6 years) who underwent laparoscopic inguinal hernia repair were prospectively evaluated with US the day before and the day after the procedure. Delayed scans were obtained at 1, 6 and 12 months. RESULTS: The laparoscopic procedure involved inversion and suturing of the hernia sac, which resulted in a nodule that plugged the internal inguinal ring and resembled a rosebud laparoscopically. US detected the 'rosebud' in all cases on the first postoperative day. Initially appearing as a rounded or ovoid mass with a slightly hypoechoic texture, the 'rosebud' became progressively smaller, more hypoechoic and more lobulated on follow-up. It disappeared in all but two cases at 6 months and in all cases at 1 year. There was no US evidence of recurrence. CONCLUSIONS: The 'rosebud' formed by this laparoscopic procedure displays typical US features and temporal changes. PMID- 12898068 TI - Effect of environmental conditions on aggregation and fibril formation of barstar. AB - The dependence on environmental conditions of the assembly of barstar into amyloid fibrils was investigated starting from the nonnative, partially folded state at low pH (A-state). The kinetics of this process was monitored by CD spectroscopy and static and dynamic light scattering. The morphology of the fibrils was visualized by electron microscopy, while the existence of the typical cross-beta structure substantiated by solution X-ray scattering. At room temperature, barstar in the A-state is unable to form amyloid fibrils, instead amorphous aggregation is observed at high ionic strength. Further destabilization of the structure is required to transform the polypeptide chain into an ensemble of conformations capable of forming amyloid fibrils. At moderate ionic strength (75 mM NaCl), the onset and the rate of fibril formation can be sensitively tuned by increasing the temperature. Two types of fibrils can be detected differing in their morphology, length distribution and characteristic far UV CD spectrum. The formation of the different types depends on the particular environmental conditions. The sequence of conversion: A-state-->fibril type I-->fibril type II appears to be irreversible. The transition into fibrils is most effective when the protein chain fulfills particular requirements concerning secondary structure, structural flexibility and tendency to cluster. PMID- 12898070 TI - Clinical quiz. Extralobar sequestration, esophageal bronchus (bronchopulmonary foregut malformation). PMID- 12898075 TI - Malignant peripheral nerve sheath tumours in neurofibromatosis type 1: MRI supports the diagnosis of malignant plexiform neurofibroma. AB - Plexiform neurofibroma (PNF) is a typical feature of neurofibromatosis 1 (NF1). About 10% of patients with NF1 develop malignant peripheral nerve-sheath tumours (MPNST), usually arising from PNF, and this is the major cause of poor survival. A better prognosis can be achieved if the tumours are diagnosed at an early stage. Our objective was to establish MRI criteria for MPNST and to test their usefulness in detecting early malignant change in PNF. MRI was performed on 50 patients with NF1 and nerve-sheath tumours, of whom seven had atypical pain, tumour growth or neurological deficits indicative of malignancy; the other 43 were asymptomatic. On MRI all seven symptomatic patients had inhomogeneous lesions, due to necrosis and haemorrhage and patchy contrast enhancement. In one patient, the multiplicity of confluent tumours with inhomogeneous areas in addition to central lesions did not allow exclusion of malignancy. Only three of the 43 asymptomatic patients had comparable changes; the other 40 patients had tumours being of relatively homogeneous structure on T1- and T2-weighted images before and after contrast enhancement. All three asymptomatic patients with inhomogeneous lesions were shown to have MPNST. PMID- 12898076 TI - Serial diffusion-weighted imaging in a patient with MELAS and presumed cytotoxic oedema. AB - A patient with mitochondrial myopathy, encephalopathy, lactic acidosis and stroke like episodes (MELAS) was studied with serial diffusion-weighted MRI (DWI) after stroke-like episodes and the apparent diffusion coefficient (ADC) was measured in an infarct-like lesion. In the acute and subacute stages, the affected area gave high signal on DWI and its ADC was much lower than that in a normal control region. In the chronic stage, the ADC became higher than that in normal brain. We therefore suggest that the stroke-like episodes did not cause vasogenic oedema but were related to energy failure and cytotoxic oedema. PMID- 12898077 TI - Craniofacial arteriovenous metameric syndrome (CAMS) 3--a transitional pattern between CAM 1 and 2 and spinal arteriovenous metameric syndromes. AB - We report a rare case of craniofacial arteriovenous metameric syndrome (CAMS) 3 arteriovenous malformations of the mandible, left VIII nerve and petrous bone. The patient, a 19-year-old girl, presented with profuse gingival bleeding during a dental procedure and we diagnosed CAMS 3 during a pre-embolisation angiogram. The distribution of the vascular lesions suggests that CAMS 3 is intermediate CAMS 1 and 2 and spinal arteriovenous metameric syndrome (SAMS). PMID- 12898078 TI - MR spectroscopy in a cervical abscess. AB - MR spectroscopy (MRS) has been used to analyse noninvasively tissues at a molecular level. Hydrogen and phosphorus MRS have been used for characterisation of intracranial solid and cystic masses, gynaecological tumours and lymph nodes. We report a cystic, tick-walled mass in the soft tissues of the neck. Single voxel proton MRS revealed a prominent acetate peak at 1.92 ppm and a diagnosis of abscess was suggested. At operation a pyogenic neck abscess was confirmed, with culture of the pus. PMID- 12898080 TI - Effect of ketoconazole on venlafaxine plasma concentrations in extensive and poor metabolisers of debrisoquine. AB - OBJECTIVE: To study the influence of CYP3A4 inhibition by ketoconazole on the disposition of venlafaxine in individuals with different CYP2D6 pheno- and genotypes. METHODS: In an open two-phase study, 21 healthy volunteers with known CYP2D6 pheno- and genotype [14 extensive metabolisers (EMs), 7 poor metabolisers (PMs)] were given a single oral dose of venlafaxine (50 mg to EMs and 25 mg to PMs). Plasma and urine levels of venlafaxine and its three metabolites were measured and the pharmacokinetics of venlafaxine were determined. After a 2-week washout period, subjects were treated for 2 days with ketoconazole (100 mg twice daily) starting 1 day before the administration of venlafaxine; and the same parameters as for the administration of venlafaxine only were measured. RESULTS: Data were evaluated from 20 subjects (14 EMs and 6 PMs) who completed the study. The dose-corrected AUC of venlafaxine was on average 2.3 times higher ( P<0.01) and that of its active metabolite O-desmethylvenlafaxine 3.4 times lower ( P<0.0001) in PMs than EMs. There was a good correlation between the debrisoquine metabolic ratio and the ratio between the AUC of venlafaxine and that of O desmethylvenlafaxine ( Rs=0.93, P<0.002). The majority of subjects showed higher plasma levels of venlafaxine and O-desmethylvenlafaxine upon co-administration of ketoconazole. AUC of venlafaxine significantly increased by 36% and that of O desmethylvenlafaxine by 26% ( P<0.01). C(max) values increased by 32% and 18%, respectively. The elimination half-life of venlafaxine was unaltered. Three of the PMs displayed marked increases in AUC (81, 126 and 206%) and C(max) (60, 72, 119%) of venlafaxine while the other three showed small or no changes. CONCLUSIONS: Ketoconazole consistently affected the disposition of venlafaxine in EMs of debrisoquine while the response in PMs was erratic. The precise mechanisms underlying this interaction remain to be elucidated. PMID- 12898081 TI - A naturalistic cohort study on effectiveness, safety and usage pattern of an over the-counter nicotine patch. Cohort study on smoking cessation. AB - INTRODUCTION. Nicotine replacement therapies (NRT) are effective for smoking cessation. After having received over-the-counter (OTC) status in Germany, concerns grew about effectiveness, increased risks, especially of adverse cardiovascular reactions, and inappropriate use of NRT. Thus, a pharmacy-based cohort study was launched. OBJECTIVES. To assess effectiveness, safety and appropriateness of use of an OTC nicotine patch (Nicotinell, Novartis Ltd.). Every customer who bought an OTC Nicotinell patch was eligible. All data were collected by self-administered questionnaires at weeks 2, 4, 8, 12 and 24 after inclusion. Six hundred and thirty-three customers were admitted, median duration of smoking was 19 years. Of the participants, 6% smoked up to 10 cigarettes per day, 43.6% between 11 and 20, 34.3% between 21 and 30, and 16.1% more than 30 cigarettes. Twenty-four weeks later, 351 participants replied: 28% (177 of 633) had quit smoking completely. Considering replies only the proportion of complete responders raised to 50.4%. There were no serious adverse events reported; 62.9% complied with the directions for use and did not use the patch for more than 3 months. About 45% smoked simultaneously with NRT. Pharmacy-based cohort studies are feasible. This study indicates that the nicotine patch is effective and safe in an OTC setting. There is still room to improve compliance with the directions for use. PMID- 12898082 TI - Rofecoxib interaction with oral anticoagulant acenocoumarol. PMID- 12898083 TI - The role of pharmacotherapy in assisting smoking cessation. AB - Tobacco use represents the single most important preventable health hazard world wide. Its consumption confers substantially increased mortality and morbidity due to cardiovascular, cerebrovascular and respiratory disease, and malignancy in a number of sites. A key goal towards improving public health in developing and developed countries alike is to reduce tobacco use across populations. Despite this, current data suggest that global tobacco use is increasing. There have been recent pharmacological advances that offer the hope of achieving more effective tobacco cessation and reduced consumption among established tobacco users. These are discussed in further detail, with reference to evidence supporting their use in clinical practice. PMID- 12898085 TI - Letter to the editor concerning Ufer et al.: Eur J Clin Pharmacol 58:779-783 (2003). PMID- 12898086 TI - A haptic size-contrast illusion affects size perception but not grasping. AB - Object features (e.g. size, shape and orientation) are relevant for recognition and identification, but also for the control of manual actions. Converging evidence suggests a dissociation between the visual systems that mediate object perception and object-directed action. Here we present evidence suggesting that a similar dissociation might exist in the haptic domain. We demonstrate that a haptic variation of a size-contrast illusion influences the perceived size of a target object, but not the degree to which the hand is opened when that object is the target of a grasping movement. This finding is consistent with the view that object perception is "scene-based" and takes into consideration not only the size of the target object but also the sizes of other nearby objects. In contrast, the control of object-directed action is primarily driven by the absolute size of the target object independent of the relative sizes of other objects in the environment, suggesting a "actor-based" frame of reference. The present findings suggest that dissociations between action and perception are not unique to the visual system, but might instead reflect a general organizational principle of sensory processing. PMID- 12898087 TI - End posture selection in manual positioning: evidence for feedforward modeling based on a movement choice method. AB - Although it is well known that the postures adopted at the ends of movements depend on where one starts and how one moves, it is not yet clear whether those differing end postures are selected before movements begin. Two experiments were designed to test the hypothesis that end postures for positioning movements are chosen before movements commence. The experiments were further designed to check whether movements are internally simulated before overt movements occur and end postures are still being selected. To address these questions we used a movement choice method. Participants were presented with two possible end postures and were asked to choose between them by moving to the one that seemed easier to adopt. End-posture choices were affected by starting positions and also by the movements that would have to be made, as affected by having obstacles in the way. The results suggest that participants relied on feedforward modeling of prospective movements as they selected end postures prior to overt movement production. The fact that the movement choice method could confirm this suggests that the method holds considerable promise as a tool for investigating motor planning. PMID- 12898088 TI - The role of vision on hand preshaping during reach to grasp. AB - During reaching to grasp objects with different shapes hand posture is molded gradually to the object's contours. The present study examined the extent to which the temporal evolution of hand posture depends on continuous visual feedback. We asked subjects to reach and grasp objects with different shapes under five vision conditions (VCs). Subjects wore liquid crystal spectacles that occluded vision at four different latencies from onset of the reach. As a control, full-vision trials (VC5) were interspersed among the blocked vision trials. Object shapes and all VCs were presented to the subjects in random order. Hand posture was measured by 15 sensors embedded in a glove. Linear regression analysis, discriminant analysis, and information theory were used to assess the effect of removing vision on the temporal evolution of hand shape. We found that reach duration increased when vision was occluded early in the reach. This was caused primarily by a slower approach of the hand toward the object near the end of the reach. However, vision condition did not have a significant effect on the covariation patterns of joint rotations, indicating that the gradual evolution of hand posture occurs in a similar fashion regardless of vision. Discriminant analysis further supported this interpretation, as the extent to which hand posture resembled object shape and the rate at which hand posture discrimination occurred throughout the movement were similar across vision conditions. These results extend previous observations on memory-guided reaches by showing that continuous visual feedback of the hand and/or object is not necessary to allow the hand to gradually conform to object contours. PMID- 12898089 TI - GTP gamma S increases Nav1.8 current in small-diameter dorsal root ganglia neurons. AB - Tetrodotoxin-resistant (TTX-R) sodium current in small-size dorsal root ganglia (DRG) neurons is upregulated by prostaglandin E(2) and serotonin through a protein kinase A (PKA)/protein kinase (PKC) pathway, suggesting G protein modulation of one or more TTX-R channels in these neurons. Recently, GTP(gammaS), a hydrolysis-resistant analogue of GTP, was shown to increase the persistent current produced by the TTX-R Na(v)1.9. In this study, we investigated the modulation of another TTX-R channel, Na(v)1.8, by GTP(gammaS) in small-diameter DRG neurons from rats using whole-cell voltage clamp recordings. Because it has been suggested that fluoride, often used in intracellular recording solutions, may bind to trace amounts of aluminum and activate G proteins, we recorded Na(v)1.8 currents with and without intracellular fluoride, and with the addition of deferoxamine, an aluminum chelator, to prevent fluoride-aluminum binding. Our results show that GTP(gammaS) (100 micro M) caused a significant increase in Na(v)1.8 current (67%) with a chloride-based intracellular solution. Although the inclusion of fluoride instead of chloride in the pipette solution increased the Na(v)1.8 current by 177%, GTP(gammaS) further increased Na(v)1.8 current by 67% under these conditions. While the effect of GTP(gammaS) was prevented by pretreatment with H7 (100 micro M), a non-selective PKA/PKC inhibitor, the fluoride-induced increase in Na(v)1.8 current was not sensitive to H7 (100 micro M), or to inclusion of deferoxamine (1 mM) in the intracellular solution. We conclude that G protein activation by GTP(gammaS) increases Na(v)1.8 current through a PKA/PKC mechanism and that addition of fluoride to the pipette solution further enhances the current, but is not a confounding variable in the study of Na(v)1.8 channel modulation by G proteins independent of a PKA/PKC pathway or binding to aluminum. PMID- 12898090 TI - Hitting moving targets: a dissociation between the use of the target's speed and direction of motion. AB - Previous work has indicated that people do not use their judgment of a target's speed to determine where to hit it. Instead, they use their judgment of the target's changing position and an expected speed (based on the speed of previous targets). In the present study we investigate whether people also ignore the target's apparent direction of motion, and use the target's changing position and an expected direction of motion instead. Subjects hit targets that moved in slightly different directions across a screen. Sometimes the targets disappeared after 150 ms, long before the subjects could reach the screen. This prevented subjects from using the target's changing position to adjust their movements, making it possible to evaluate whether subjects were relying on the perceived or an expected (average) direction to guide their movements. The background moved perpendicular to the average direction of motion in some trials. This influences the target's perceived direction of motion while leaving its perceived position unaffected. When the background was stationary, subjects hit disappearing targets along their trajectory, just as they hit ones that remained visible. Moving the background affected the direction in which subjects started to move their hand, in accordance with the illusory change in direction of target motion. If the target disappeared, this resulted in a hit that was systematically off the target's trajectory. If the target remained visible, subjects corrected their initial error. Presumably they did so on the basis of information about the target's changing position, because if the target disappeared they did not correct the error. We conclude that people do use the target's perceived direction of motion to determine where to hit it. Thus the perceived direction of motion is treated differently than the perceived speed. This suggests that the motion of an object is not broken down into speed components in different directions, but that speed and direction are perceived and used separately. PMID- 12898091 TI - Evidence for recurrent inhibition of reciprocal inhibition from soleus to tibialis anterior in man. AB - Reciprocal inhibition between ankle flexors and extensors has been the subject of numerous studies in Man. They have demonstrated that this reciprocal inhibition is in all likelihood caused by a disynaptic pathway at least partly fed by Ia afferents. It is thus generally agreed that this reciprocally organized inhibition between ankle flexors and extensors in Man is similar to the reciprocal Ia inhibition described in the cat. This conclusion has, however, been challenged, when Jankowska and McCrea described in the cat a non-reciprocal group I inhibition involving interneurones co-excited by Ia and Ib afferents and mediating inhibition to both antagonistic and non-antagonistic motoneurones. The only way to distinguish between reciprocal Ia inhibition and non-reciprocal group I inhibition is to test if the inhibition is blocked by recurrent inhibition, since only Ia interneurones are inhibited by recurrent inhibition. In the present study, reciprocal inhibition from soleus to tibialis anterior was thus investigated following activation of soleus-coupled Renshaw cells in normal human subjects. It was found that reciprocal inhibition induced in tibialis anterior motoneurones by the activation of soleus group I afferents is deeply depressed by activation of soleus-coupled Renshaw cells. This finding provides the missing data to identify disynaptic inhibition between antagonistic ankle muscles as a reciprocal Ia inhibition. PMID- 12898092 TI - Nicotinic effects on excitatory field potentials recorded from the immature CA3 area of rat hippocampal slices. AB - We investigated the nicotinic modulation of the excitatory field potentials recorded from the immature (postnatal day 10-20) hippocampal CA3 area, in the presence of the GABA(A) antagonist bicuculline methiodide (BMI, 10 microM). Nicotine (50 microM) enhanced the evoked field potentials; its effects were also observed in the presence of the GABA(B) antagonist 2-hydroxy-saclofen (250 microM; added to BMI) and were blocked by pre-perfusion with the nicotinic antagonist hexamethonium (HXM, 50 microM). The potentiating effects of nicotine in BMI persisted during prolonged perfusion (more than 20 min), while those in control perfusion medium were transient. The nicotinic antagonists HXM (50 microM), methyllycaconitine (MLA, 0.01 microM) and dihydro-beta-erythroidine (DHbetaE, 50 microM) potentiated CA3-evoked field potentials. Perfusion of HXM in the presence of the anticholinesterase eserine (1 microM) or the muscarinic antagonist atropine (1 microM) did not alter its effects. None of the nicotinic agents tested changed the frequency of spontaneous BMI-induced epileptiform discharges (nicotine, HXM, MLA, DhbetaE), suggesting that nicotinic receptors do not drive spontaneous epileptiform discharges in this in vitro model. These experiments demonstrate that nicotinic receptors are activated tonically during disinhibition and modulate the activity of excitatory synapses in the immature CA3 hippocampal area. The persistent nicotinic facilitatory effects during disinhibition versus the transient in control conditions indicate that nicotinic modulation depends on environmental conditions and also that nicotinic receptors may be a contributing factor in early-life seizures. PMID- 12898093 TI - Inhibition of muscle spindle afferent activity during masseter muscle fatigue in the rat. AB - The influence of muscle fatigue on the jaw-closing muscle spindle activity has been investigated by analyzing: (1) the field potentials evoked in the trigeminal motor nucleus (Vmot) by trigeminal mesencephalic nucleus (Vmes) stimulation, (2) the orthodromic and antidromic responses evoked in the Vmes by stimulation of the peripheral and central axons of the muscle proprioceptive afferents, and (3) the extracellular unitary discharge of masseter muscle spindles recorded in the Vmes. The masseter muscle was fatigued by prolonged tetanic masseter nerve electrical stimulation. Pre- and postsynaptic components of the potentials evoked in the Vmot showed a significant reduction in amplitude following muscle fatigue. Orthodromic and antidromic potentials recorded in the Vmes also showed a similar amplitude decrease. Furthermore, muscle fatigue caused a decrease of the discharge frequency of masseter muscle spindle afferents in most of the examined units. The inhibition of the potential amplitude and discharge frequency was strictly correlated with the extent of muscle fatigue and was mediated by the group III and IV afferent muscle fibers activated by fatigue. In fact, the inhibitory effect was abolished by capsaicin injection in the masseter muscle that provokes selective degeneration of small afferent muscle fibers containing neurokinins. We concluded that fatigue signals originating from the muscle and traveling through capsaicin-sensitive fibers are able to diminish the proprioceptive input by a central presynaptic influence. In the second part of the study, we examined the central projection of the masseter small afferents sensitive to capsaicin at the electron-microscopic level. Fiber degeneration was induced by injecting capsaicin into the masseter muscle. Degenerating terminals were found on the soma and stem process in Vmes and on the dendritic tree of neurons in Vmot. This suggests that small muscle afferents may influence the muscle spindle activity through direct synapses on somata in Vmes and on dendrites of neurons in Vmot. PMID- 12898094 TI - The kinaesthetic perception of Euclidean distance: a study of the detour effect. AB - An experiment investigated the mechanisms by which humans estimate Euclidean distances on the basis of kinaesthetic cues. Blindfolded participants followed straight and curvilinear paths with a hand-held stylus (encoding phase). Then, with a straight movement, they estimated the Euclidean distance between the start and end-points of the path (response phase). The experiment contrasted an On axis condition, in which encoding and response movements were spatially aligned, and an Off-axis condition, in which they were displaced laterally. Performances were slightly more accurate in the On-axis condition than in the Off-axis condition. In both conditions, however, errors were consistently smaller when the path covered a larger surface. The results showed that small paths yielded an overestimation of the Euclidean distance, the relative errors increasing with the length of curvilinear paths. The findings are compared with results of other studies in which distances were estimated on the basis of haptic cues. PMID- 12898095 TI - Motor learning processes in a movement-scaling task in olivopontocerebellar atrophy and Parkinson's disease. AB - Nine Parkinson's disease (PD), seven olivopontocerebellar atrophy (OPCA) patients and two age-matched control groups learned a linear arm movement-scaling task over 2 days, requiring movements proportional in length to visually presented target-bars. Scaling was acquired through knowledge of results (KR concerning the direction and magnitude of errors) following every second acquisition trial. Initial acquisition of both groups was significantly worse than their respective controls (poorer movement scaling), but rapidly improved to nearly identical levels. Retention for the PD group's movement scaling was as good as controls initially, but markedly poorer after 24 h. The OPCA group did not show this deficit. Both patient groups extrapolated accurately to longer, previously unpracticed target distances (no KR provided), suggesting an unimpaired capacity to generate and use an internal representation of the movement scaling. They also rapidly learned a new scaling relationship when the gain was changed. Overall, the learning of this movement-scaling task was not adversely affected in OPCA, and the impairment was restricted primarily to longer-term retention in PD. The study suggests that: (1) the ability to acquire movement scaling in a task that requires conscious use of error feedback and no new coordination may depend little on the cerebellum, and (2) the basal ganglia may participate in longer term storage of scaling information. PMID- 12898096 TI - Differences in the abilities of individual fingers during the performance of fast, repetitive tapping movements. AB - Using 12 healthy male subjects, the dynamic motor ability of individual fingers was investigated under four different finger tapping conditions. These were: maximum speed tapping with one finger (single-finger tapping), alternate movement of two fingers (double-finger tapping), double-finger tapping in an unsupported condition, and submaximum constant speed tapping with one finger in a passive manner. Key-contact forces for all fingers and the movement velocity of the tapping finger were monitored. With the exception of the unsupported condition, non-tapping fingers were maintained in contact with designated keys during the tapping tasks. It was found that the index finger attained the fastest cadence and greatest movement velocity, followed by the middle, little and ring fingers, respectively. Subjective assessment of rank order of "difficulty" of tapping by the subjects was highly correlated with tapping cadence. Thus dynamic motor function, as indicated by rapid, repetitive movement, differs among the individual fingers. Parallel changes were observed in the key-contact force of the neighboring non-tapping fingers during tapping. The range of the non-tapping finger forces was largest during tapping by the ring finger. A similar trend was found for passive tapping, during which the magnitude of key-contact force was less than one-third of that observed during active tapping. The lower cadence achieved by the ring finger may be attributed more to a lack of independence at the level of voluntary neuromuscular control, than to innate mechanical interaction with the other fingers. Tapping cadence of each finger was lower for the double-finger mode than for the single-finger mode. The magnitude of the observed decrease in cadence during double-finger tapping was, on the other hand, strongly dependent on finger-combination. The decrease was smallest for the index middle finger-combination, and greatest for the ring-little finger-combination. Compatibilities with other fingers can play an essential role in the dynamic motor function of individual fingers. During the unsupported task, in which interactions were diminished by allowing all fingers to move freely, tapping cadence increased markedly. Therefore, the lower cadences observed in specific finger-combinations may be partly attributed to anatomical and neural interdigit interactions. PMID- 12898098 TI - Role of the neocortex in consolidation of fear conditioning memories in rats. AB - The aim of the present work was to investigate, by means of the reversible inactivation technique, the distinct roles of three neocortical sites, the Prefrontal (PFC), Frontal (FC) and Parietal (PAC) cortices, during memory consolidation of conditioned freezing to a tone, a defined conditioned stimulus (CS), and to the training context. Reversible tetrodotoxin inactivations were performed at increasing post-acquisition delays (0.25, 24 or 96 h). Retention testing was always performed 72 h after the inactivation procedure in order to avoid any possible influence on the retrieval phase. It was found that none of the three sites are involved in context freezing consolidation, but that FC and PAC are differentially involved in the consolidation of memory to the CS. FC inactivation was followed by retention impairment only when performed immediately after acquisition while PAC inactivation was followed by amnesia when performed both immediately after acquisition and 24 h later, but not when performed 96 h later. PFC inactivation was not followed by retention impairment at any of the employed delays. These findings show that some neocortical structures are involved in auditory-cued fear conditioning during the initial phases of the consolidation process. PMID- 12898097 TI - A new model of experimental parotitis in rats and its implication for trigeminal nociception. AB - A rat model of chronic parotitis was developed following a direct injection of Complete Freund's adjuvant (CFA) into the unilateral parotid gland via the parotid duct without skin incision. The nocifensive behavior, plasma extravasation in the parotid gland, and trigeminal Fos protein expression, a marker of neuronal activation, were analyzed in this model and compared to that of the saline-injected rats. A significant reduction of the escape threshold to mechanical stimulation of the lateral face on the ipsilateral side to the CFA injection was observed at 1-6 days after CFA injection as compared to that of the pre-CFA control ( P<0.01). The lateral face region contralateral to the CFA injection also showed mechanical hyperalgesia at 1-6 days after injection ( P<0.05). The plasma extravasation was significantly increased in the parotid gland ipsilateral to CFA injection as compared to that of the parotid gland with saline injection at 3 days after injection as shown by Evans' blue dye extravasation ( P<0.05). Bilateral expression of Fos protein-like immunoreactive cells was observed in the transition zone between the trigeminal spinal nucleus interpolaris (Vi) and caudalis (Vc) and paratrigeminal nucleus (Pa5). On the other hand, a significant unilateral expression of Fos protein-positive cells was observed on the ipsilateral side of the upper cervical (C2) dorsal horn ( P<0.05). This model of parotitis can be used to study trigeminal pain mechanisms associated with sialadenitis. A unique feature of this preparation is that the inflammation was limited to the parotid gland after intraductal injection of CFA, allowing analysis of peripheral input from a defined orofacial region. The model will be useful in developing new strategies to treat chronic orofacial pain. PMID- 12898099 TI - Effect of trans-bullar gentamicin treatment on guinea pig angular and linear vestibulo-ocular reflexes. AB - This study provides the first systematic examination of the effects of intratympanic gentamicin instillation on vestibulo-ocular responses of guinea pigs during both Earth-vertical yaw axis and off-vertical axis rotation. A scleral search coil was sutured to the right eye of pigmented female guinea pigs prior to trans-bullar instillation of a 0.2-ml bolus of either 20 mg/ml or 40 mg/ml of gentamicin (1) into the right middle ear (unilateral treatment groups) or (2) into both ears (bilateral treatment groups). Two weeks later, eye movement responses were tested during yaw axis sinusoidal rotation at 7 frequencies (0.02, 0.05, 0.1, 0.2, 0.5, 1 and 2 Hz, 40 deg/s peak velocity) and during off-vertical axis rotation at five constant velocities (20, 40, 60, 80 and 100 deg/s), tilted 30 deg relative to the earth-vertical axis. The main result was that unilateral trans-bullar gentamicin instillation produced almost exclusively unidirectional deficits in horizontal angular vestibulocular reflex (HVOR) responses and modulation and bias responses to off-vertical axis rotation (OVAR). The HVOR gain was reduced during rotation toward the injected ear in a dose-dependent manner for frequencies of 1 Hz and lower, but there was no effect on responses during rotation toward the intact ear. Further, the modulation and bias responses to OVAR were reduced profoundly in a dose-dependent manner during rotation toward the treated ear. It is suggested that these effects indicate selective cytotoxic and/or physiologic effects of gentamicin intoxication in the inner ear or, possibly, the vestibular nerve and central nervous system. PMID- 12898100 TI - Responses of gerbil utricular afferents to translational motion. AB - In the present study, we report the sensitivity of utricular afferents to sinusoidal translational motion in the horizontal plane. The head orientation was altered relative to the direction of translational travel in 30 degrees increments to allow determination of the head orientation that elicited the maximal and minimal responses of each afferent neuron. We determined gain and phase relationships at a constant peak linear acceleration of 0.1 g applied at frequencies between 0.20 and 2.0 Hz for multiple head orientations. The response dynamics and vector of maximal sensitivity for the utricular afferents are consistent with those reported for other mammalian species. Irregularly (CV>0.3) and intermediate (0.1 or =100 ms. We found that auditory thalamus bursts occur in synchronized electroencephalogram states (SWS and anesthesia), but also during W. Although the burst characteristics did not differ among the three states, group data showed that the proportion of bursts within spike trains was the greatest under anesthesia. This observation resulted from two types of effects: (1) the percentage of non bursting cells was lowest under anesthesia and (2) some cells under anesthesia exhibited up to 90% of bursts, whereas during W or SWS the highest proportion of bursts did not exceed 40%. The presence of these bursts is discussed with regards to the known fluctuations of membrane potential which occur in these various states. PMID- 12898104 TI - Anodic stripping voltammetric determination of mercury using multi-walled carbon nanotubes film coated glassy carbon electrode. AB - An electrochemical method for the determination of trace levels of mercury based on a multi-walled carbon nanotubes (MWNT) film coated glassy carbon electrode (GCE) is described. In 0.1 mol L(-1) HCl solution containing 0.02 mol L(-1) KI, Hg(2+) was firstly preconcentrated at the MWNT film and then reduced at -0.60 V. During the anodic potential sweep, reduced mercury was oxidized, and then a sensitive and well-defined stripping peak at about -0.20 V appeared. Under identical conditions, a MWNT film coated GCE greatly enhances the stripping peak current of mercury in contrast to a bare GCE. Low concentrations of I(-) remarkably improve the determining sensitivity, since this increases the accumulation efficiency of Hg(2+) at the MWNT film coated GCE. The stripping peak current is proportional to the concentration of Hg(2+) over the range 8 x 10(-10) 5 x 10(-7) mol L(-1). The lowest detectable concentration of Hg(2+) is 2 x 10( 10) mol L(-1) at 5 min accumulation. The relative standard deviation (RSD) at 1 x 10(-8) mol L(-1) Hg(2+) was about 6% ( n=10). By using this proposed method, Hg(2+) in some water samples was determined, and the results were compared with those obtained by atomic absorption spectrometry (AAS). The two results are similar, suggesting that the MWNT-film coated GCE has great potential in practical analysis. PMID- 12898105 TI - Flow injection on-line ultrasound-assisted extraction of iron in meat samples coupled to a flame atomic absorption spectrometric system. AB - Iron was extracted on-line from solid meat samples by a simple and rapid continuous ultrasound-assisted extraction system (CUES). The CUES is connected to a flow injection manifold, which allows the on-line flame atomic absorption spectrometric determination of iron. A Plakett-Burman design was used for the optimisation of the CUES. The method achieved a total sampling frequency of 11 samples per hour with a relative standard deviation for the complete procedure of 0.4%. The detection limit was 0.6 microg g(-1) (dry mass) for a sample amount of 30 mg. Accurate results were obtained by measuring the certified reference materials BCR-186 (pig kidney) and BCR-184 (bovine muscle). The analytical procedure was applied to different real meat samples with satisfactory results. PMID- 12898106 TI - An integrated electrochemical fructose biosensor based on tetrathiafulvalene modified self-assembled monolayers on gold electrodes. AB - An integrated amperometric fructose biosensor based on a gold electrode (AuE) modified with a self-assembled monolayer (SAM) of 3-mercaptopropionic acid (MPA) on which fructose dehydrogenase (FDH) and the mediator tetrathiafulvalene (TTF) are co-immobilized by cross-linking with glutaraldehyde is reported. Variables concerning the behavior of the biosensor were optimized by taking the slope value obtained for the fructose calibration plot in the 0.1-1.0 mM concentration range as a criterion of selection. At an applied potential of +0.20 V, a good repeatability of such slope values (RSD=6.7%, n=10) was obtained with no need to apply a cleaning or pretreatment procedure to the modified electrode. Moreover, results from five different TTF-FDH-MPA-AuEs yielded a RSD of 5.8%. The useful lifetime of one single biosensor was approximately 30 days, exhibiting a 93% of the original response on the 33rd day. A linear calibration graph was obtained for fructose over the 1.0 x 10(-5)-1.0 x 10(-3) M range, with a limit of detection of 2.4 x 10(-6) M. The effect of potential interferents was evaluated. The TTF-FDH-MPA-AuE also performed well in the flow-injection mode. The biosensor was used for the determination of fructose in real samples, and the results compared with those provided by using a commercial enzyme test kit. PMID- 12898107 TI - Determination of methylmercury in environmental samples using static headspace gas chromatography and atomic fluorescence detection after aqueous phase ethylation. AB - A rapid and automated method for the determination of monomethylmercury (MMHg) in environmental samples was developed using headspace gas chromatography with atomic fluorescence detection in combination with aqueous phase ethylation. Sample preparation steps were optimized for sediments, biological samples, and water samples using certified reference materials and real samples with a broad range of MMHg concentrations. Different extraction procedures were compared for both sediments and biological samples. The methods were applied in the intercomparison exercises for the certification of MMHg in sediments (IAEA 405) and in Oyster tissue (BCR 710) and the results were accepted for certification. The detection limits for MMHg are 0.002 ng Hg/g for sediments and biological samples and 0.01 ng Hg/L for water samples. The method was tested for methylation artifacts; no artifact was observed in the sediment samples and CRMs tested. PMID- 12898108 TI - Olfactory receptors: molecular basis for recognition and discrimination of odors. AB - The daunting task of our nose to detect and discriminate among thousands of low molecular-weight organic compounds with diverse chemical structures and properties requires an enormous molecular recognition capacity. This is based on distinct proteins, capable of recognizing and binding odorous compounds, including odorant-binding proteins, which are supposed to shuttle odorous compounds through the nasal mucus, and most notably the odorant receptors, which are heptahelical membrane proteins coupling via G-proteins onto intracellular transduction cascades. From more than a thousand genes each olfactory neuron is supposed to express only one receptor subtype. Receptors appear to be selective but rather non-specific-i.e. a distinct odorant activates multiple receptors and individual receptors respond to multiple odorants. It is the molecular receptive range of its receptor type which determines the reaction spectrum of a sensory neuron. Populations of cells equipped with the same receptor type project their axons to common glomeruli, thereby transmitting the molecular receptive range of a receptor type into the receptive field of glomerulus. Recent insight into the molecular basis of odor recognition and the combinatorial coding principles of the olfactory system may provide some clues for the design and development of technical sensors, electronic noses. In this review more emphasis has been placed on physiological rather than analytical aspects. PMID- 12898109 TI - Design of luminescent biochips based on enzyme, antibody, or DNA composite layers. AB - The use of beads bearing bioactive molecules to develop generic biochips based on chemi- and electro-chemiluminescent detection was evaluated. The biochips were composed of arrayed biosensors, including enzyme-charged beads, antigen-charged beads, or oligonucleotide-charged beads, entrapped in poly(vinyl alcohol) (PVA SbQ) photopolymer. In each case the sensing layers were spotted at the surface of a glassy carbon electrode as 0.3 microL drops, generating 500-800 microm spots. The luminescent reactions were either catalysed by horseradish peroxidase or triggered by application of a +850 mV potential between the glassy carbon electrode and a platinum pseudo-reference. Enzyme biochips were designed for the concomitant detection of choline, glucose, glutamate, lactate, lysine, and urate, based on the corresponding oxidase-charged beads and the electro-chemiluminescent (ECL) reaction with luminol-immobilised beads of the hydrogen peroxide produced. Limits of detection of 1 micromol L(-1) for glutamate, lysine and uric acid, 20 micromol L(-1) for glucose, and 2 micromol L(-1) for choline and lactate were found with detection ranging over three decades at least. Use of the electro chemiluminescent biochip was extended to a tri-enzymatic sensing layer based on kinase-oxidase activity for detection of acetate. A reaction sequence using acetate kinase, pyruvate kinase, and pyruvate oxidase enabled the production of H2O2 in response to acetate injection in the range 10 micromol L(-1) to 100 mmol L(-1). Based on IgG-bearing beads, a chemiluminescent immuno-biochip has been also realised for the model detection of human IgG. Biotin-labelled anti-human IgG were used in a competitive assay, in conjunction with peroxidase-labelled streptavidin. Free antigen could then be detected with a detection limit of 25 pg (10(8) molecules) and up to 15 ng. In a similar way, the use of oligonucleotide immobilised beads enabled the realisation of DNA-sensitive biochips which could be used to detect a biotin-labelled sequence al a level of 5x10(8) molecules. PMID- 12898110 TI - Square-wave voltammetric determination of cefoperazone in a bacterial culture, pharmaceutical drug, milk, and urine. AB - Use of square-wave voltammetry (SWV) for determination of cefoperazone (CFPZ) in some buffers, bacterial culture, urine, and milk is described. CFPZ provides a specific voltammetric signal which is affected by pH and solution components. Determination of CFPZ in Britton-Robinson buffer, pH 4.4, is sensitive with a low detection limit (about 0.5 nmol L(-1)). In a more complex medium (bacterial 2YT medium, pH 7.2) the detection limit was approximately 1.5 micromol L(-1). We provide evidence that SWV is a suitable and quick method for CFPZ determination in a culture of living bacteria without separation of biomass. We have found big differences between methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-sensitive Staphylococcus aureus (MSSA) in cultivation in the presence of CFPZ, depending on time. When CFPZ is cleaved by penicillinase, a new SWV peak b appears at more positive potentials. This peak rises both with increasing concentration of enzyme and with cleavage time while the original CFPZ peak is simultaneously decreasing. We determined the concentration of CFPZ in the drug Pathozone by the standard addition method and achieved good agreement with the declared value of CFPZ in the drug. With a simple pretreatment procedure it is possible to determine CFPZ in milk; for urine no pretreatment was required. Using SWV we could detect CFPZ concentrations as low as 500 nmol L(-1) in bovine milk and human urine. PMID- 12898111 TI - Amperometric and spectrophotometric determination of carbaryl in natural waters and commercial formulations. AB - The work presented describes the development and evaluation of two flow-injection analysis (FIA) systems for the automated determination of carbaryl in spiked natural waters and commercial formulations. Samples are injected directly into the system where they are subjected to alkaline hydrolysis thus forming 1 naphthol. This product is readily oxidised at a glassy carbon electrode. The electrochemical behaviour of 1-naphthol allows the development of an FIA system with an amperometric detector in which 1-naphthol determination, and thus measurement of carbaryl concentration, can be performed. Linear response over the range 1.0 x 10(-7) to 1.0 x 10(-5) mol L(-1), with a sampling rate of 80 samples h(-1), was recorded. The detection limit was 1.0 x 10(-8) mol L(-1). Another FIA manifold was constructed but this used a colorimetric detector. The methodology was based on the coupling of 1-naphthol with phenylhydrazine hydrochloride to produce a red complex which has maximum absorbance at 495 nm. The response was linear from 1.0 x 10(-5) to 1.5 x 10(-3) mol L(-1) with a detection limit of 1.0 x 10(-6) mol L(-1). Sample-throughput was about 60 samples h(-1). Validation of the results provided by the two FIA methodologies was performed by comparing them with results from a standard HPLC-UV technique. The relative deviation was <5%. Recovery trials were also carried out and the values obtained ranged from 97.0 to 102.0% for both methods. The repeatability ( RSD, %) of 12 consecutive injections of one sample was 0.8% and 1.6% for the amperometric and colorimetric systems, respectively. PMID- 12898112 TI - Liquid chromatography-mass spectrometry approach for the characterization and purification of crude synthetic peptide hormones. AB - In this study, conditions for the optimal separation by LC-UV and characterisation by LC-ES-MS of crude mixtures generated during SPPS of several peptide hormones are compiled. The linear solvation energy relationship (LSER) methodology has been used to predict the retention of the target peptides and their side products and then to develop a separation LC methodology with applicability on both the analytical and preparative scale. Identification of these side products by LC-ES-MS analysis has been made on the basis of their calculated molecular masses. This method may be regarded as a key tool for the optimisation of the synthetic procedures and for complying with regulatory agencies' requirements before commercialisation of a safe and effective peptide based pharmaceutical drug. PMID- 12898113 TI - Evaluations of solid electrodes for use in voltammetric monitoring of heavy metals in samples from metallurgical nickel industry. AB - Evaluation of different solid electrode systems for detection of zinc, lead, cobalt, and nickel in process water from metallurgical nickel industry with use of differential pulse stripping voltammetry has been performed. Zinc was detected by differential pulse anodic stripping voltammetry (DPASV) on a dental amalgam electrode as intermetallic Ni-Zn compound after dilution in ammonium buffer solution. The intermetallic compound was observed at -375 mV, and a linear response was found in the range 0.2-1.2 mg L(-1) (r(2)=0.98) for 60 s deposition time. Simultaneous detection of nickel and cobalt in the low microg L(-1) range was successfully performed by use of adsorptive cathodic stripping voltammetry (AdCSV) of dimethylglyoxime complexes on a silver-bismuth alloy electrode, and a good correlation was found with corresponding AAS results (r(2)=0.999 for nickel and 0.965 for cobalt). Analyses of lead in the microg L(-1) range in nickel plating solution were performed with good sensitivity and stability by DPASV, using a working electrode of silver together with a glassy carbon counter electrode in samples diluted 1:3 with distilled water and acidified with H(2)SO(4) to pH 2. A new commercial automatic at-line system was tested, and the results were found to be in agreement with an older mercury drop system. The stability of the solid electrode systems was found to be from one to several days without any maintenance needed. PMID- 12898114 TI - Direct spectrometric determination of proteins in body fluids using a near infrared cyanine dye. AB - A new near-infrared (NIR) dye, 1,1'-disulfobutyl-3,3,3',3' tetramethylindotricarbocyanine (DTCY) has been developed for the quantitation of proteins in solution. The method is based on the binding of DTCY to proteins under acidic conditions. The binding of DTCY to proteins causes a new band at 814 nm. The maximum binding number of bovine serum albumin (BSA) with DTCY was measured as 100. The linear range is 0.3-40 microg mL(-1) for BSA and human serum albumin (HSA), respectively. Except for Fe(2+), Cu(2+), and cetyltrimethylammonium bromide, all of the examined coexisting substances show no interference in the assay. The method has been applied to the quantitation of proteins in serum and urine with recoveries between 96 and 105%. PMID- 12898115 TI - On-line monitoring of a frontal chromatographic separation using inductively coupled plasma atomic emission spectroscopy. AB - A test array is described employing a destructive analytical technique for the long-term monitoring of an industrial-scale separation process. As an example, we chose frontal chromatography as the separation and ICP/AES as the analytical method. The feed solution of the process was conveyed by a process pump via the separation unit to a sample station, where a small portion was diverted and transported by a roller pump into the spectrometer. We equipped our array with different loops for operating the process, calibrating the instrument and verifying the calibration. We obtained identical results for the different loops by absorbing the pulsation of the process pump and arranging for identical suction lines of the spectrometer pump. Based on the results, we redesigned the sample station for a technical application using only commercially available parts. PMID- 12898116 TI - Partition and location of nimesulide in EPC liposomes: a spectrophotometric and fluorescence study. AB - Study of the mechanism of action of anti-inflammatory drugs (NSAIDs) and their side-effects may fall in the domain of membranology. In this work the extent of the interaction between an NSAID (nimesulide) and membrane phospholipids was quantified by the partition coefficient, K(p ), using egg phosphatidylcholine (EPC) liposomes as cell membrane models. The liposome/aqueous phase partition coefficients were determined under physiological conditions, by derivative spectrophotometry and fluorescence quenching. Derivative spectrophotometry allows elimination of background signal effects (light scattering) due to the presence of liposomes. Theoretical models, accounting for simple partition of the NSAID between two different media, were used to fit the experimental data, allowing the determination of K(p)in multilamellar vesicles (MLVs) and large unilamellar vesicles (LUVs). The location of nimesulide in MLVs and LUVs was evaluated by fluorescence quenching using spectroscopic probes located at different sites on the membrane. All n-AS probes were quenched and the relative quenching efficiencies were ordered as 2-AS<6-AS approximately 9-AS<12-AS; this suggests the drug is deeply buried in the membrane. Fluorescence quenching using the 12-AS probe was also used to determine the partition coefficient of the drug in MLVs and LUVs. The two techniques yield similar results. Finally, measurement of zeta potential in the presence of different concentrations of nimesulide was performed to investigate possible changes in the zwitterionic phosphatidylcholine membranes. The membrane surface potential was not altered, which seems to be an indication that nimesulide binds to lipid bilayer mostly by hydrophobic interactions. PMID- 12898117 TI - Miniaturized dynamic liquid-liquid extraction of organophosphate triesters from blood plasma using the hollow fibre-based XT-tube extractor. AB - A miniaturized liquid-liquid extractor for bioanalytical sample preparation is described. The extractor consists of a polypropylene hollow fibre mounted inside polytetrafluoroethylene (PTFE) tubing by means of a cross (X) connector and a tee (T) connector. All parts are commercially available, inexpensive, and easily assembled. The aqueous sample, injected into a carrier flow, is pumped along the outside of the fibre and the organic phase, which also wets the pores of the hollow fibre wall, is pumped inside. Eight organophosphate triester (OPE) plasticisers/flame retardants were extracted from 50 microL spiked blood plasma that had been mixed with 50 microL formic acid to denature plasma proteins. The organic phase was a mixture of hexane and methyl tert-butyl ether (MTBE). A high concentration of formic acid in the sample and of MTBE in the organic phase had positive effects on the recovery of some OPE. When investigating the recovery as a function of extraction time it was found that the extraction reached a maximum after 10 min, at a flow-rate of 15 microL min(-1). Recoveries varied between 40 and 80% with RSD around 4% for most compounds. The whole 150- microL extract was injected into a GC-MS system equipped with a programmed-temperature vaporization (PTV) injector. With the MS in selected-ion monitoring (SIM) mode, the LOD for triphenyl phosphate and 2-ethylhexyl diphenyl phosphate were 0.3 and 0.2 ng mL( 1), respectively. More than 40 plasma extractions were performed with the same fibre without any detectable change in extraction efficiency. PMID- 12898118 TI - An uncertainty budget for trace analysis by isotope-dilution ICP-MS with proper consideration of correlation. AB - Isotope-dilution mass spectrometry (IDMS) is considered to be a method without significant correction factors. It is also believed that this method is well understood. But unfortunately a large number of different uncertainty budgets have been published that consider different correction factors. These differences lead to conflicting combined uncertainties especially in trace analysis. It is described how the known correction factors must be considered in the uncertainty budget of values determined by IDMS combined with ICP-MS (ICP-IDMS). The corrections applied are dead time, background, interference, mass discrimination, blank correction and air buoyancy.IDMS measurements consist always of a series of isotope abundance ratio measurements and can be done according to different measurement protocols. Because the measurement protocols of IDMS are often rather sophisticated, correlations of influence quantities are difficult to identify. Therefore the measurement protocol has to be carefully considered in the specification of the measurand and a strategy is presented to properly account for these correlations. This will be exemplified for the estimation of mass fractions of platinum group elements (PGEs) and Re in the geological reference material UB-N (from CRPG-CNRS, Nancy in France) with ICP-IDMS. The PGEs with more than one isotope and the element Re are measured with on-line cation-exchange chromatography coupled to a quadrupole ICP-MS. All contents are below 10 microg kg(-1). Only osmium is separated from the matrix by direct sparging of OsO(4) into the plasma. This leads to transient signals for all PGEs and Re. It is possible to estimate the combined uncertainties and keep them favourably small despite the low contents, the transient signals and the sophisticated correction model. PMID- 12898119 TI - Determination of sulfur and selected trace elements in metallothionein-like proteins using capillary electrophoresis hyphenated to inductively coupled plasma mass spectrometry with an octopole reaction cell. AB - The determination of sulfur in biologically relevant samples such as metalloproteins is described. The analytical methodology used is based on robust on-line coupling between capillary electrophoresis (CE) and octopole reaction cell inductively-coupled plasma mass spectrometry (ORC-ICP-MS). Polyatomic ions that form in the plasma and interfere with the determination of S at mass 32 are minimised by addition of xenon to the collision cell. The method has been applied to the separation and simultaneous element-specific detection of sulfur, cadmium, copper, and zinc in commercially available metallothionein preparations (MT) and metallothionein-like proteins (MLP) extracted from liver samples of bream ( Abramis brama L.) caught in the river Elbe, Germany. Instrumental detection limits have been calculated according to the German standard procedure DIN 32645 for the determination of sulfur and some simultaneously measured trace elements in aqueous solution. For sulfur detection limits down to 1.3 microg L(-1) ((34)S) and 3.2 microg L(-1) ((32)S) were derived. For the other trace elements determined simultaneously detection limits ranging from 300 ng L(-1) ((58)Ni) to 500 ng L(-1) ((66)Zn, (55)Mn) were achieved. For quantification of sulfur and cadmium in a commercially available MT preparation under hyphenated conditions the use of external calibration is suggested. Finally, the need for proper sample preparation technique will be discussed. PMID- 12898120 TI - Antagonism of the antidepressant-like effects of clenbuterol by central administration of beta-adrenergic antagonists in rats. AB - RATIONALE: Stimulation of central beta(2) adrenergic receptors produces antidepressant-like effects on behavior. At present, it is not known what brain sites are involved in mediating such effects, although some recent evidence suggests the importance of the dorsal hippocampus. OBJECTIVE: Experiments were carried out to determine whether central administration of beta-adrenergic antagonists blocks antidepressant-like effects produced by peripheral administration of the beta(2)-adrenergic agonist clenbuterol. METHODS: The following were determined: 1) the ability of ICV or intrahippocampal administration of the non-selective beta adrenergic antagonists propranolol and CGP-12177, which are lipophilic and hydrophilic, respectively, to antagonize the effects of peripherally administered clenbuterol on differential-reinforcement-of low-rate (DRL) behavior; 2) the effects of clenbuterol, administered bilaterally into the dorsal hippocampus, on DRL behavior. RESULTS: The antidepressant-like effects of clenbuterol, i.e. reduced response rate and increased reinforcement rate under the DRL schedule, were antagonized by either ICV or bilateral intrahippocampal infusions of propranolol or CGP-12177; CGP-12177 was approximately 8-fold more potent than propranolol. Direct infusion of clenbuterol into the bilateral dorsal hippocampus also produced antidepressant-like effects. CONCLUSIONS: Central beta-adrenergic receptors, in particular those in the dorsal hippocampus, are involved in the mediation of the antidepressant-like effect of clenbuterol. Probably resulting from its enhanced access to the sites of action, the hydrophilic antagonist CGP-12177 was more potent than the lipophilic antagonist propranolol, even though they exhibit similar potency in vitro. PMID- 12898121 TI - Effects of continuous nicotine infusion on nicotine self-administration in rats: relationship between continuously infused and self-administered nicotine doses and serum concentrations. AB - RATIONALE: The efficacy of nicotine replacement therapy (NRT) for smoking cessation is limited. One reason for this limited efficacy may be that typical serum nicotine concentrations provided by NRT do not match the peak arterial nicotine concentrations achieved from smoking. OBJECTIVE: The purpose of the present study was to determine whether continuous nicotine infusion at a rate producing serum nicotine concentrations that match the estimated peak arterial nicotine concentrations associated with nicotine self-administration (NSA) in rats produces greater suppression of NSA than lower infusion rates. METHODS: The effects of continuous nicotine infusion were studied by intravenously administering nicotine at various rates (1.0, 3.0, and 8.0 mg/kg per day) to rats concurrently self-administering nicotine (0.03 mg/kg per infusion) during 23-h sessions or cocaine (0.17 mg/kg per infusion) during 2-h sessions. RESULTS: Continuous nicotine infusion suppressed NSA in a rate-related fashion. NSA was suppressed by 17, 50, and 73% at infusion rates of 1.0, 3.0 and 8.0 mg/kg per day, respectively. The 8.0-mg/kg per day infusion rate, which provided venous serum nicotine concentrations equaling the peak arterial concentrations associated with NSA, suppressed NSA to a greater extent than lower infusion rates. The 8.0-mg/kg per day nicotine infusion rate had no effect on cocaine maintained responding, demonstrating that its effects were specific for suppression of NSA. This infusion rate provided a mean percentage replacement of nicotine from NSA of more than 700%. Reacquisition of NSA after suppression by the two highest infusion rates was delayed compared with reacquisition after saline extinction. CONCLUSIONS: Continuous nicotine infusion produced an infusion rate-related suppression of NSA that was greatest when the infusion provided nicotine doses and venous serum concentrations substantially higher than those typically associated with NRT in humans. PMID- 12898122 TI - Haloperidol and clozapine antagonise amphetamine-induced disruption of latent inhibition of conditioned taste aversion. AB - RATIONALE: Latent inhibition (LI) describes a process by which repeated pre exposure of a stimulus without any consequence retards the learning of subsequent conditioned associations with that stimulus. It is well established that LI is impaired in rats and in humans by injections of the indirect dopamine agonist amphetamine (AMPH), and that this disruption can be prevented by co administration of either the typical neuroleptic haloperidol (HAL) or the atypical neuroleptic clozapine (CLZ). OBJECTIVES: Most of what is known of the pharmacology of LI is derived from studies using either the conditioned emotional response or the conditioned active avoidance paradigm. The goal of the present study was to determine whether these results would generalize to the conditioned taste aversion assay. METHODS: We tested whether AMPH (0.5 mg/kg) pretreatment would disrupt LI of a conditioned aversion to sucrose, and if so, which stage of the procedure is critical for mediating the disruption; in addition, we tested whether HAL (0.2 mg/kg) or CLZ (5.0 mg/kg) could restore such an expected LI disruption. RESULTS: We determined that AMPH disrupted LI when it was injected before pre-exposure and prior to conditioning, but not if the rats were injected before either stage alone. When HAL or CLZ was given 40 min before AMPH (before both pre-exposure and conditioning), it blocked LI disruption. CONCLUSION: These results are in line with the pharmacology of LI as derived from other conditioning paradigms. We conclude that the pharmacological regulation of LI in the CTA paradigm is similar to what has been observed previously in the conditioned emotional response and the conditioned active avoidance paradigms. PMID- 12898123 TI - Effects of NRA0045, a novel potent antagonist at dopamine D4, 5-HT2A, and alpha1 adrenaline receptors, and NRA0160, a selective D4 receptor antagonist, on phencyclidine-induced behavior and glutamate release in rats. AB - RATIONALE: Administration of phencyclidine (PCP) to animals produces abnormal behavior such as hyperlocomotion, stereotyped behavior, and ataxia; this abnormal behavior is only weakly blocked by dopamine D(2) receptor antagonists. This study examined the effects of a novel thiazole derivative, NRA0045 which potently antagonizes not only dopamine D(4) receptors but also 5-HT(2A) and alpha(1) adrenaline receptors, and NRA0160, a selective dopamine D(4) receptor antagonist, on PCP-induced abnormal behavior, and accompanying increases in extracellular levels of glutamate in the medial prefrontal cortex. Furthermore, this study compared the effects of these drugs with those of clozapine and haloperidol. METHODS: To study the effects of NRA-drugs, atypical and typical antipsychotics, we measured locomotor activity with an infra-red sensor, and stereotypy and ataxia on a rating scale. Extracellular glutamate levels were measured by in vivo microdialysis. RESULTS: NRA0045 (1 or 3 mg/kg) or clozapine (1 mg/kg) attenuated hyperlocomotion, stereotypy, and ataxia induced by PCP (7.5 mg/kg) without affecting behavior after saline injection. Although haloperidol (0.1 or 1 mg/kg) attenuated or inhibited PCP-induced behavior, this drug also affected behavior after saline injection. NRA0160 (0.1, 1, or 3 mg/kg) had no effect on behavior induced by PCP or saline. NRA0045 (3 mg/kg), but not NRA0160, inhibited PCP induced increases in glutamate levels in the medial prefrontal cortex. PCP induced hyperlocomotion correlated with the PCP-induced increases in glutamate levels in this brain region. CONCLUSIONS: These results suggest that the effects of NRA0045 on PCP-induced abnormal behavior are similar to those of the atypical antipsychotic clozapine. NRA0045 probably attenuates PCP-induced abnormal behavior by inhibiting the PCP-induced increase in glutamate levels in the medial prefrontal cortex; this inhibition may be mediated via the blockade of 5-HT(2A) receptors. PMID- 12898124 TI - Castration in rats impairs performance during acquisition of a working memory task and exacerbates deficits in working memory produced by scopolamine and mecamylamine. AB - RATIONALE: Although much research has focused on the effects of ovarian hormones on learning and memory in females, less information is available regarding the effects of testicular hormones on learning and memory in males. Additionally, despite evidence of an interaction of testicular hormones and the cholinergic system in areas of the brain implicated in learning and memory, no information is available regarding the behavioral consequences of that interaction. OBJECTIVES: We assessed the effect of castration in male rats on working memory during acquisition of a radial maze. We also assessed the interactive effects of castration and scopolamine, a muscarinic receptor antagonist, as well as mecamylamine, a nicotinic receptor antagonist, on behavior. METHODS: Young adult male rats were castrated or underwent sham surgeries. Beginning 10 days after surgeries, performance on a task of working memory was assessed across 24 days of acquisition in an eight-arm radial maze. Following acquisition, scopolamine and mecamylamine dose-effect curves were established. RESULTS: Castration of male rats significantly decreased arm-choice accuracy during acquisition. Castration significantly exacerbated impairments in arm-choice accuracy produced by scopolamine as well as mecamylamine, without altering the disruptive effects of the drugs on the rate at which rats entered the arms of the maze. CONCLUSIONS: These results indicate that castration in male rats impairs working memory during acquisition of a spatial maze task. Additionally, these results suggest that the absence of testicular hormones increases the sensitivity of male rats to the impairing effects of scopolamine as well as mecamylamine on working memory. PMID- 12898125 TI - Neurophysiological evidence for abnormal cognitive processing of drug cues in heroin dependence. AB - RATIONALE: Recent studies provide evidence for specific aspects of cue processing in addictive disorders. OBJECTIVE: The present study employs event related potentials (ERPs) to investigate heroin related visual information processing. METHODS: Neutral and heroin related pictures were presented to 19 male abstinent heroin dependent patients and 14 male healthy controls. RESULTS: Patients exhibited larger slow positive wave (SPW) components of the ERP on heroin related pictures than on neutral pictures. Within healthy control subjects there was no difference on the SPW between neutral and heroin pictures. Within heroin dependent patients, mean SPW response to heroin pictures was correlated with post experiment craving. CONCLUSION: This study provides neurophysiological evidence that information processing of drug-related information is abnormal in heroin dependent patients. The results provide further evidence for the cognitive and neurobiological accounts of substance dependence such as the incentive sensitization theory. PMID- 12898126 TI - The role of nitric oxide in systemic and hepatic haemodynamics in the rat in vivo. AB - The physiological role of nitric oxide (NO) in portal venous and hepatic arterial haemodynamics in the rat in vivo during healthy and diseased conditions remains unclear. The present study determined the physiological role of nitric oxide in hepatic haemodynamics in the rat in vivo during healthy conditions as a basis for future pharmacological work. Male Wistar rats (300-350 g) were anaesthetised with fentany/fluanisone (0.3 mg/kg s.c.) and midazolam (0.3 mg/kg s.c.) and heparinised (30 U/100 g i.v.) via a cannulated left carotid artery for measurement of heart rate, mean arterial pressure, and the difference between systolic and diastolic blood pressures (P(S-D)). Following laparotomy, two distal ileocolic veins were cannulated, one catheter introduced to a distance of 1 cm and used for intraportal drug injections and the other to the main trunk of the portal vein for continuous measurement of portal venous pressure. The portal venous trunk and hepatic artery were carefully isolated and electromagnetic probes placed around each of them for measurement of portal venous flow and hepatic arterial flow. Augmentation of NO production was achieved by intraportal injection of 0.2, 0.4, 0.6 and 0.8 g/kg L-arginine and the NO donor, 3 morpholinosydnonimine (SIN-1), was injected intraportally at 0.2, 0.4, 0.6 and 0.8 mg/kg. L-NAME, the non-selective NOS inhibitor, was injected intraportally in increasing doses of 5, 10, 15 and 20 mg/kg in the absence or presence of L arginine in doses of 0.2 and 0.5 g/kg. L-arginine increased portal blood flow by 25% without significant changes in systemic haemodynamics. SIN-1 decreased mean arterial pressure by 33% with no effect on portal blood flow. Both L-arginine and SIN-1 reduced portal venous pressure by 25% in a dose-dependent manner. L-NAME had no effect on portal haemodynamics despite a significant increase in systemic arterial pressure of 60% that was reduced dose-dependently by L-arginine. Hepatic arterial flow increased by 88% and 49% at the second and third doses of L arginine and by 68% and 27% at the first two doses of L-NAME. No significant changes in hepatic arterial flow were found when L-NAME and L-arginine were given together. It is concluded that augmented endogenous NO production increased portal flow. Inhibition of endogenous NO had no effect on portal haemodynamics. Endogenous NO may not play a major role in regulation of portal haemodynamics in the rat in vivo. PMID- 12898127 TI - The effects of systemically administered taurine and N-pivaloyltaurine on striatal extracellular dopamine and taurine in freely moving rats. AB - The second most abundant cerebral amino acid, taurine, is widely consumed in the so-called "energy drinks". Therefore, its possible actions on the brain are of great interest. In the present experiments taurine was given intraperitoneally to rats in order to study if it can be administered systemically in large enough amounts to alter cerebral dopaminergic transmission or to induce hypothermia. In addition, the effects of subcutaneously administered lipophilic taurine analogue, N-pivaloyltaurine, were studied. The extracellular striatal taurine and dopamine concentrations were estimated using in vivo microdialysis in awake and freely moving rats, and the rectal temperatures were measured. Taurine at the total dose of 45 mmol/kg i.p. led to a maximally 8-fold increased striatal extracellular taurine concentration, induced a long-lasting hypothermia, and significantly reduced the striatal extracellular dopamine concentration. The latter effect was strengthened by co-treatment with reuptake inhibitor nomifensine. N pivaloyltaurine (15 mmol/kg in total, s.c.) only slightly elevated the striatal extracellular taurine concentration, failed to alter the rectal temperature, and in contrast to taurine somewhat elevated the striatal extracellular dopamine concentration suggesting a different mechanism or locus of action from that of taurine. Finally, our experiments using brain microdialysis confirmed the earlier findings that taurine is slowly eliminated from the brain. The results clearly indicate that systemically given taurine enters the brain in concentrations that induce pharmacological effects. PMID- 12898129 TI - Hexachlorobenzene impairs glucose metabolism in a rat model of porphyria cutanea tarda: a mechanistic approach. AB - Hexachlobenzene (HCB), one of the most persistent environmental pollutants, induces porphyria cutanea tarda (PCT). The aim of this work was to analyze the effect of HCB on some aspects of glucose metabolism, particularly those related to its neosynthesis in vivo. For this purpose, a time-course study on gluconeogenic enzymes, pyruvate carboxylase (PC), phosphoenolpyruvate carboxykinase (PEPCK), glucose-6-phosphatase (G-6-Pase) and on pyruvate kinase (PK), a glycolytic enzyme, was carried out. Plasma glucose and insulin levels, hepatic glycogen, tryptophan contents, and the pancreatic insulin secretion pattern stimulated by glucose were investigated. Oxidative stress and heme pathway parameters were also evaluated. HCB treatment decreased PC, PEPCK, and G 6-Pase activities. The effect was observed at an early time point and grew as the treatment progressed. Loss of 60, 56, and 37%, respectively, was noted at the end of the treatment when a considerable amount of porphyrins had accumulated in the liver as a result of drastic blockage of uroporphyrinogen decarboxylase (URO-D) (95% inhibition). The plasma glucose level was reduced (one-third loss), while storage of hepatic glucose was stimulated in a time-dependent way by HCB treatment. A decay in the normal plasma insulin level was observed as fungicide intoxication progressed (twice to four times lower). However, normal insulin secretion of perifused pancreatic Langerhans islets stimulated by glucose during the 3rd and 6th weeks of treatment did not prove to be significantly affected. HCB promoted a time-dependent increase in urinary chemiluminiscence (fourfold) and hepatic malondialdehide (MDA) content (fivefold), while the liver tryptophan level was only raised at the longest intoxication times. These results would suggest that HCB treatment does not cause a primary alteration in the mechanism of pancreatic insulin secretion and that the changes induced by the fungicide on insulin levels would be an adaptative response of the organism to stimulate gluconeogenesis. They showed for the first time that HCB causes impairment of the gluconeogenic pathway. Therefore, the reduced levels of glucose would thus be the consequence of decreased gluconeogenesis, enhanced glucose storage, and unaffected glycolysis. The impairment of gluconeogenesis (especially for PEPCK) and the related variation in glucose levels caused by HCB treatment could be a consequence of the oxidative stress produced by the fungicide. Tryptophan adds its effect to this decrease in the higher phases of HCB intoxication, where its levels overcome the control values possibly owing to the drastic decline of URO D. This derangement of carbohydrates leads porphyric hepatocytes to have lower levels of free glucose. These results contribute to our understanding of the protective and modulatory effect that diets rich in carbohydrates have in hepatic porphyria disease. PMID- 12898132 TI - Characterization of early deaths of non- Saccharomyces yeasts in mixed cultures with Saccharomyces cerevisiae. AB - The survival of Kluyveromyces thermotolerans and Torulaspora delbrueckii in mixed cultures with Saccharomyces cerevisiae was examined at low oxygen availability in a defined grape juice medium. In these fermentations, K. thermotolerans and T. delbrueckii died off earlier than S. cerevisiae, and K. thermotolerans and T. delbrueckii exhibited parabolic death kinetics. Furthermore, the early deaths seemed to be non-apoptotic in nature. In order to understand the mechanism causing the early deaths, various single- and mixed-culture fermentations were carried out. The early deaths could not be explained by nutrient depletion or the presence of toxic compounds. Rather, they seemed to be mediated by a cell-to-cell contact mechanism at high cell densities of S. cerevisiae, and to a lesser ability of K. thermotolerans and T. delbrueckii to compete for space, as compared to S. cerevisiae. These results contribute to an increased understanding of why K. thermotolerans and T. delbrueckii die off before S. cerevisiae in wine fermentations. PMID- 12898128 TI - Magnolol induces apoptosis in vascular smooth muscle. AB - Magnolol, an active component extracted from Magnolia officinalis, has various pharmacological effects, including potent antioxidant activity. In the present study, we investigated the effect of magnolol on apoptosis in rat vascular smooth muscle cells (VSMCs), using terminal-deoxynucleotidyl-transferase-mediated deoxyuridine triphosphate nick end labelling (TUNEL) and flow cytometric analysis. Magnolol (5-20 micro M) concentration-dependently induced significant VSMC apoptosis, this effect being blocked by the caspase inhibitor benzyloxycarbonyl-Val-Ala-Asp-fluoromethylketone (z-VAD-fmk, 50 micro M). To study the molecular mechanism, the mitochondrial death pathway was examined. Magnolol increased caspase-3 and caspase-9 activities significantly and reduced the mitochondrial potential (Deltapsi(m)). The levels of B-cell leukaemia/lymphoma-2 (Bcl-2), but not those of Bcl-2-associated X protein (Bax) or Bcl-x(L), were down-regulated concentration dependently by magnolol. In an animal model, balloon angioplasty-induced neointima formation was inhibited significantly by magnolol and Bcl-2 protein levels were reduced. Taken together, these results show that magnolol induces apoptosis in VSMCs via the mitochondrial death pathway. This effect is mediated through down-regulation of Bcl-2 protein levels, both in vivo and in vitro. Magnolol thus shows potential as a novel therapeutic agent for the treatment of atherosclerosis and re-stenosis. PMID- 12898133 TI - Gene structure and transcriptional regulation specific to the groESL operon from the psychrophilic bacterium Colwellia maris. AB - The groESL operon of a psychrophilic bacterium, Colwellia maris, was cloned and sequenced. The operon contains two ORFs of 291 bp and 1,650 bp separated by 210 bp. Northern blot analysis suggested that the groESL operon was transcribed as a bicistronic mRNA, and that the amount of mRNA markedly increased after the temperature was raised from 10 degrees C to 20 degrees C. Although the optimum temperatures for GroESL function are different in psychrophilic, mesophilic, and thermophilic bacteria, the deduced amino acid sequences of C. maris GroES and GroEL showed remarkably high similarity with those of GroES and GroEL from mesophilic and thermophilic bacteria. A putative promoter similar to the Escherichia coli sigma(32) consensus sequence was identified. One specific feature of C. maris groESL was that in the putative untranslated region the G+C content was about 24 mol%, which is much lower than that of mesophilic bacteria such as E. coli. The low G+C content may be important for maintaining transcription at low temperatures. PMID- 12898134 TI - Agrobacterium- tumefaciens-mediated transformation of Helminthosporium turcicum, the maize leaf-blight fungus. AB - Agrobacterium tumefaciens has the ability to transfer its T-DNA to plants, yeast, filamentous fungi, and human cells and integrate it into their genome. Conidia of the maize pathogen Helminthosporium turcicum were transformed to hygromycin B resistance by a Agrobacterium- tumefaciens-mediated transformation system using a binary plasmid vector containing the hygromycin B phosphotransferase ( hph) and the enhanced green fluorescent protein ( EGFP) genes controlled by the gpd promoter from Agaricus bisporus and the CaMV 35S terminator. Agrobacterium- tumefaciens-mediated transformation yielded stable transformants capable of growing on increased concentrations of hygromycin B. The presence of hph in the transformants was confirmed by PCR, and integration of the T-DNA at random sites in the genome was demonstrated by Southern blot analysis. Agrobacterium- tumefaciens-mediated transformation of Helminthosporium turcicum provides an opportunity for advancing studies of the molecular genetics of the fungus and of the molecular basis of its pathogenicity on maize. PMID- 12898135 TI - Poly beta-hydroxybutyrate depolymerase (PhaZ) in Azospirillum brasilense and characterization of a phaZ mutant. AB - Like many other prokaryotes, rhizobacteria of the genus Azospirillum produce high levels of poly-beta-hydroxybutyrate (PHB) under sub-optimal growth conditions. Utilization of PHB by bacteria under stress has been proposed as a mechanism that favors their compatible establishment in competitive environments. PHB depolymerase (PhaZ) is an essential enzyme in PHB degradation. The phaZ gene was identified in Azospirillum brasilense, cloned, sequenced, and shown to be located on the chromosome. Insertion of a kanamycin-resistant cassette within phaZ of A. brasilense resulted in a phaZ mutant that was unable to degrade PHB; however, carbon source utilization was similar in both the wild-type and the mutant strain. The ability of the wild-type to endure starvation conditions, ultraviolet irradiation, heat, and osmotic shock, and to grow in the presence of hydrogen peroxide was higher than that of the mutant strain. By contrast, the ability of the phaZ mutant strain to endure desiccation was higher than that of the wild type strain. No differences between the strains were seen in their ability to endure sonication, or to survive in carrier materials used for soil inoculants. In addition, motility was the same between the two strains, whereas cell aggregation and exopolysaccharide production were higher in the wild-type than in the phaZ mutant strain. PMID- 12898136 TI - Deformation characteristics of the human mandible in low impact experiments. AB - A total of 11 human mandibles were subjected to physical impacts under standardised conditions. Two impact sites and directions were tested and the impact load was varied in four steps. Two occlusional strengths were applied and the influence of simulated soft tissue covering was recorded. The deformation of the bones was measured using strain gauge strips located at eight defined sites. In a series of frontal to occipital impacts the frontal areas and the collum showed the largest length changes. Increasing impact intensities led to a proportional increase of the length changes. An increase of the occlusional strength was either protective (at the collum) or it increased the deformation (frontal area). The soft tissue covering was only partly protective. Lateral impact was characterised by a compression on the side of the impact and stretching on the other side only. The intensity and speed of deformation increased with increasing distance from the site of impact. A fixed occlusion caused an increase of compression at the site of impact and an increasing stretching in the frontal part of the bone and at the opposite collum. PMID- 12898137 TI - Characterization of eight Y-STR loci and haplotypes in a Chinese Han population. AB - In this study we analyzed the eight Y-STR loci, DYS443, DYS444, DYS448, DYS453, DYS455, DYS456, DYS457 (DYS437) and DYS458, investigated haplotype distributions of these Y-STR loci in a Chinese Han population, and sequenced alleles of the eight loci for clarifying the structure. Extracted DNA was amplified by PCR and the PCR products were analyzed by non-denaturing horizontal polyacrylamide gel electrophoresis with a discontinuous buffer system. Alleles were sequenced on an ABI 3700 using a Dye Terminator Cycle sequencing kit. DYS443, DYS453, DYS455 and DYS456 were found to be simple repeat systems, while DYS444, DYS448, DYS457 (DYS437) and DYS458 were complex repeat systems. The gene diversities of DYS443, DYS444, DYS448, DYS453, DYS455, DYS456, DYS457 (DYS437) and DYS458 were 0.7742, 0.7671, 0.7453, 0.3545, 0.0549, 0.6988, 0.6148 and 0.8213, respectively. The haplotype diversity for 8 Y-STR loci was 0.9996, and the discrimination capacity was 0.9815. The results indicate that these eight loci are useful Y-linked markers for forensic applications. PMID- 12898138 TI - Clinical analysis of extranodal non-Hodgkin's lymphoma in the sinonasal tract. AB - We investigated the clinical analysis of non-Hodgkin's lymphoma (NHL) of the sinonasal tract, including the survival rate and treatment outcome. Fifty patients who had previously received a diagnosis of extranodal NHL of the sinonasal cavity from May 1992 to April 2001 were included. We reviewed the patients' clinical characteristics and the survival rates, retrospectively. Of 50 patients, 49 were classified as having extranodal NK/T cell lymphoma and only one patient as having diffuse large B cell (DLBC) lymphoma according to the new WHO classification. Even though higher mortality rates were observed in patients receiving chemotherapy alone than in those receiving chemotherapy and radiation therapy in the advanced stage, the combination treatment of chemotherapy and radiation therapy failed to demonstrate a significantly higher survival rate. PMID- 12898140 TI - Post-tonsillectomy haemorrhage: reusable and disposable instruments compared. AB - The aim of this study was to determine if the introduction of disposable instruments for tonsillectomy resulted in a significant change in post tonsillectomy haemorrhage rates. This is a prospective comparative study of haemorrhage rates for cold dissection (CD) tonsillectomy in adults using reusable instruments during 1999-2000 ( n=83) and disposable instruments between August and December 2001 ( n=111). Haemorrhage rates in children with reusable instruments ( n=156) and disposable instruments ( n=115) were also compared. Confidence intervals were established for the differences between study groups along with exact levels of significance. No difference was found in the overall reactionary haemorrhage rate [ P=0.32, Diff 0.9% (95% CI; -3.2 to +0.4)] or secondary haemorrhage rate [ P=1.00, Diff 3.4% (95% CI; -0.09 to +0.01)] between reusable and disposable instruments. The introduction of disposable instruments has not produced a statistically significant increase in post-tonsillectomy haemorrhage rates in our centre. PMID- 12898139 TI - Immunomodulation of tissue-engineered transplants: in vivo bone generation from methylprednisolone-stimulated chondrocytes. AB - Subcutaneously implanted, in vitro engineered tissue is generally affected by the immune system of the host even in autogenous transplantation. The aim of this study was to investigate immunomodulation of subcutaneously implanted tissue engineered cartilage transplants by intramuscular methylprednisolone application. Transplants consisted of auricular rabbit chondrocytes, polylactide-polyglycolide co-polymer fleeces and species-specific fibrin or agarose. The transplants were subcutaneously implanted in the ridge. Thereafter, animals were separated into two groups, one with and one without methylprednisolone treatment. The specimens were histologically investigated after 6 and 12 weeks. Fleece fiber degradation was complete after 12 weeks, and all transplants showed areas of calcification. The corticosteroid-treated group presented pronounced trabecular bone generation without fibrous tissue infiltration. The untreated group showed sporadic islets of calcification without coherent bone formation, and adjacent fibrous tissue had infiltrated the transplants. Native controls and corticoid-treated transplants did not exhibit bone generation or signs of fibrous tissue infiltration. This study found that immunomodulation by intramuscular methylprednisolone application protects tissue-engineered autogenous chondrocyte transplants from fibrous tissue infiltration and induces trabecular bone formation. PMID- 12898141 TI - Choanal polyp originating from the middle turbinate. AB - Choanal polyps usually originate from the maxillary sinus. Unusual origins such as the sphenoid sinus, ethmoid sinus, nasal septum and hard and soft palate have been reported in the literature. Herein, we report a rare case of choanal polyp originating from the middle turbinate that was removed by an endoscopic surgery technique. The computed tomographic findings are described and the literature is reviewed. PMID- 12898142 TI - Spontaneous fetal heart rate deceleration: an ominous sign for fetal outcome. AB - BACKGROUND: The aim of this study was to establish the prognostic import of spontaneous fetal heart-rate deceleration, a sign of fetal distress. METHODS: This retrospective study enrolled 169 patients with fetal heart-rate deceleration treated at Shin-Kong Wu Ho Su Memorial Hospital from 1 January 1998 to 24 November 2000. Reviewed variables included type of fetal heart-rate irregularity (including early, variable, late, and spontaneous variants), neonatal outcome, Doppler results, gestational age and weight at birth, and amniotic fluid index. RESULTS: The spontaneous form of fetal heart-rate deceleration was determined for 11 of the 169 patients (6.5%) diagnosed with fetal heart-rate deceleration, with a 5-min Apgar score below 7 observed for 5 (45.5%) of these subjects, and neonatal complications for 9 (81.8%). Mean gestational age was 32.2+/-3.8 weeks; mean birth weight was 1,560+/-723.9 g. Spontaneous deceleration has a sensitivity of 29%, specificity of 99%, positive predictive value of 81.8%, and negative predictive value of 86.1%. CONCLUSION: Although of the different fetal heart-rate deceleration types the spontaneous variant has the lowest incidence rate, the complication and mortality rates are highest and it is also associated with lower birth weight and higher incidence of pre-term labor. Failure of the fetus to cope with the compromised placental flow and uterine compression resulting from oligohydramnios may be fundamental to the etiology of spontaneous fetal-heart rate deceleration. Once diagnosed, Doppler ultrasound may be used to assess placental-vessel flow. Emergency treatment may be mandatory if deterioration is noted. PMID- 12898143 TI - Identification of a mutant allele of the androgen receptor gene in a family with androgen insensitivity syndrome: detection of carriers and prenatal diagnosis. AB - We report the results of a molecular study of a large family segregating the complete form of the Androgen Insensitivity Syndrome (CAIS) in several family members from three generations. We identified the mutant allele by polymerase chain reaction (PCR) amplification of the short tandem repeat (CAG)n, highly polymorphic in the population, present in the first exon of the androgen receptor (AR) gene. In this family four different alleles were detected and one of these showed a perfect segregation with the disease. This study enabled us to identify the heterozygous females in this family. We think that this simple, indirect test, is also suitable for prenatal diagnosis of Morris' syndrome when the mother is heterozygous for the size of the short tandem repeat and one affected subject in the family may be studied. PMID- 12898144 TI - Semen parameters in patients with unilateral testicular cancer compared to patients with other malignancies. AB - AIM: The aim of our study was to evaluate pretreatment semen quality in patients with unilateral testicular cancer in comparison to semen parameters of patients suffering from other malignancies. METHODS: Semen analysis according to WHO recommendations were performed in 16 patients suffering from unilateral testicular cancer (group 1) and in 21 patients who were diagnosed to have other malignant tumors (group 2: non-Hodgkin's lymphoma: n=7, Hodgkin's disease: n=6, leukemia: n=3, osteosarcoma: n=2, rectal cancer: n=2, thyroid carcinoma: n=1) prior to anti-cancer treatment. RESULTS: Normozoospermia was observed in none of the testicular cancer patients but in five of the other patients ( p=0.047). Sperm concentration was significantly decreased in the patients of group 1 compared to those of group 2 (18.7+/-22.3x10(6)/ml versus 35.6+/-31.3x10(6)/ml, p=0.03). The percentage of rapid progressive motile spermatozoa was significantly less in testicular cancer patients than in patients suffering from other malignancies (WHO group a: 1.1+/-2.0% versus 4.7+/-5.6%, p=0.02). Normal morphological findings of spermatozoa were found in a smaller percentage in group 1 (16.2+/-6.0% versus 26.1+/-18.0%, p=0.03). CONCLUSIONS: Our investigations revealed that especially testicular cancer has an adverse influence on sperm quality. Hence, we advise careful and sufficient cryopreservation of semen in those patients taking the negative impact of cytotoxic treatments on semen parameters into account. PMID- 12898145 TI - Retrorectal endometrial cyst. AB - CASE REPORT: An unusual case of retrorectal endometriosis is presented. This presented as an asymptomatic pelvic mass at the level of the pelvic floor not contiguous with the pelvis or peritoneal cavity. RESULT: It was successfully excised via an abdominal approach. PMID- 12898146 TI - Adhesion molecules, activin and inhibin--candidates for the biochemical prediction of hypertensive diseases in pregnancy? AB - BACKGROUND: The aim of the prospective study was to compare standard parameters as Doppler ultrasound and 24-h blood pressure measurement with possible maternal serological markers regarding their prognostic value in predicting hypertensive diseases in pregnancy. MATERIALS: Twenty-four-hour blood pressure measurement was performed before and after 32+0 gestational week in 57 pregnant women with either chronic hypertension ( n=13), preeclampsia ( n=21), pregnancy-induced hypertension (PIH; n=12) or normotension ( n=11). Blood samples were taken and the concentrations of soluble intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), activin A and inhibin A were determined as well as serum uric acid, creatinine, total serum protein and serum albumin. Doppler ultrasound of the uterine arteries was examined before 32+0 gestational week in the same patients. For the statistical evaluation Kruskal Wallis-Test and Mann-Whitney-U-Test were performed. Differences in the predictive value were evaluated by receiver-operating characteristics. RESULTS: VCAM-1 was significantly elevated in women developing hypertensive diseases as compared to normotensive women (preeclampsia: p<0.001; PIH: p<0.05; chronic hypertension: p<0.001). In early pregnancy activin A and inhibin A were significantly higher in preeclamptic patients than in the other groups (activin A: normotension: p<0.005; PIH: p<0.001; chronic hypertension: p<0.005) (inhibin A: normotension: p<0.005; PIH: p<0.001; chronic hypertension: p<0.01), thus suggesting them to be specific markers for the development of preeclampsia. Mean arterial pressure was significantly elevated in preeclampsia ( p<0.001) and chronic hypertension ( p<0.005) as compared to normotensives. CONCLUSION: Twenty-four-hour blood pressure monitoring with determination of mean arterial pressure and measurement of VCAM-1, activin A and inhibin A as serum parameters can be suggested as useful tests in the specific prediction of different types of hypertensive diseases in pregnancy. PMID- 12898147 TI - Oxidizability of apolipoprotein B-containing lipoproteins, levels of lipid peroxidation products and antioxidants in normal pregnancy. AB - OBJECT: The object was to assess oxidizability of apolipoprotein B-containing lipoproteins, levels of lipid peroxidation products and antioxidants in normal pregnancy. METHOD: Serum malondialdehyde, vitamin E, carotenoids, albumin, uric acid, bilirubin, triglyceride, total cholesterol levels and in vitro copper induced oxidation of apolipoprotein B-containing lipoproteins were investigated in 21 healthy pregnant and 22 nonpregnant women. Mann-Whitney U test was used for the statistical analyses. RESULTS: Serum malondialdehyde, vitamin E, triglyceride and total cholesterol levels were significantly higher; albumin and uric acid levels and vitamin E/triglyceride + total cholesterol and carotenoids/triglyceride + total cholesterol ratios were significantly lower in the pregnant group. Basal malondialdehyde concentrations of apolipoprotein B containing lipoprotein fraction and Delta-malondialdehyde values determined at 30, 60, 90, 120, 150 and 180th minutes of incubation were significantly lower in the pregnant group compared to those in the non-pregnant group. CONCLUSION: Although greater lipid peroxidation product levels reflect an enhanced lipid peroxidation status in normal pregnancy, apolipoprotein B-containing lipoprotein fraction of normal pregnant women compared with the nonpregnant subjects seems to be protected from oxidation. PMID- 12898148 TI - Expression and localization of tight junction-associated proteins in human hair follicles. AB - Tight junctions (TJ) are barrier-forming intercellular junctions selectively sealing cells and controlling the paracellular pathway. They have been well characterized in simple epithelia and endothelia but have only recently been described in stratified epithelia such as epidermis, oesophagus and oral mucosa. Various epithelial layers which are partly in morphogenic continuity with the epidermis and develop therefrom during early fetal life, build the human hair follicle. The barrier function of these epithelial layers seems to be important for the universal continuity of the barrier represented by the skin. We show the presence of the TJ proteins ZO-1, occludin, and various claudins in the hair follicle and demonstrate their impressive heterogeneous distribution pattern within a given stratum as well as within its different epithelial layers. Coexpression of the various TJ proteins, arguing for typical TJ structures, can be observed especially in cell layers facing the hair shaft and the stratum corneum, and in addition at the border between the outer and inner root sheaths. Usually they are found in close proximity to desmosomal and adherens junction proteins. The morphological and biological importance of these findings and the possible roles of TJ in hair follicles, e.g. in follicular penetration, are discussed. PMID- 12898149 TI - The mesenchymal substrate influences the epithelial phenotype in a three dimensional cell culture. AB - Cell-matrix interactions are thought to influence epithelial structure, growth, and differentiation. Three-dimensional cell cultures were used to study the effects of the composition of the dermal equivalent on the morphology of epithelium grown from HaCaT skin keratinocytes. Three commercial preparations, a basement membrane preparation from a tumor (matrix 1), two preparations consisting of collagen type I and III (matrix 3 and 4) and a noncommercial preparation containing collagen type I (matrix 2) were investigated. The effects of fibroblasts of different origin (vaginal mucosa, oral buccal mucosa, and skin) were investigated in connection with matrix 4. The histomorphology and expression of the proteins PCNA, Ki-67, p53, p21, pankeratin, involucrin, cytokeratin 10 (Ck10), Ck17, Ck19 and collagen type IV were evaluated. Three-dimensional cultures of HaCaT cells gave rise to an epithelium with an immature and hyperproliferative character, showing active proliferation with intense PCNA staining. Both matrix 1 and matrix 2 resulted in an epithelium with budding into the matrix and some degree of layering. These epithelia showed only scattered Ck17 and Ck19 expression but a low terminal differentiation potential as indicated by scattered Ck10 and involucrin staining. The epithelia of cocultures with matrices 3 and 4 were positive for Ck17 and Ck19. However, the epithelium on matrix 3 showed strong expression of the terminal differentiation marker Ck10 and involucrin. This methodological study provides evidence of the importance of standardization of the composition of the matrix to avoid confounding effects on epithelial morphology and protein expression in studies using a three-dimensional epithelial culture model. PMID- 12898150 TI - Human VAT-1: a calcium-regulated activation marker of human epithelial cells. AB - BACKGROUND AND AIMS: Human VAT-1 (hVAT-1) is a homologue of the synaptic vesicle membrane protein of Torpedo californica. Its coding gene is located near the BRCA1 locus and thus hVAT-1 may be linked to an inherited predisposition to breast and ovary cancer. However, the hVAT-1 protein expression pattern in normal epithelial tissues such as skin, mammary gland and ovary, as well as in tumours of the mammary gland and ovary, has not been studied. METHODS: To address this issue, an immunohistological analysis of biopsies of normal epidermis and lesional epidermis of bullous pemphigoid and pemphigus vulgaris patients was undertaken. RESULTS: hVAT-1-expression was observed in basal keratinocytes of lesional epidermis of bullous pemphigoid patients but not in normal epidermis or in lesional epidermis of pemphigus vulgaris patients. Moreover, hVAT-1 expression in HaCaT cells was found to be calcium-dependent. Normal and malignant mammary and ovary epithelium were found to be hVAT-1-negative. CONCLUSIONS: Our results indicate that hVAT-1 exerts a specific function in keratinocyte physiology, in particular in calcium-regulated processes, with no evident deregulation in malignancies of the breast and ovary. PMID- 12898153 TI - Cerebral amyloid angiopathy in the elderly: vessel walls changes and relationship with dementia. AB - Abeta peptide deposits are observed in brain cortical and leptomeningeal microvessels in a few families, in patients with Alzheimer's disease and in cognitively normal elderly subjects. These deposits, which cause Abeta amyloid angiopathy, are usually associated with other lesions induced by Abeta peptide and tau pathologies. To investigate the consequences of cerebral amyloid angiopathy on arterial morphology and search for correlations with the degree of cognitive impairment, we carried out a prospective clinicopathological and morphometric study in 29 institutionalized elderly patients cognitively normal or affected with sporadic dementia associated with Alzheimer-type lesions, cerebral infarcts or both. We measured the external and internal diameters of arteries 40 120 microm wide, containing moderate or severe Abeta deposits, and of unaffected arteries in the temporal and frontal lobes. We found no differences in the mean external diameters. In contrast, the mean internal diameters of vessels with moderate Abeta deposits were smaller than those of unaffected vessels. Conversely, the internal diameters of severely affected vessels were larger than those of unaffected vessels. This suggests that arterial walls become thicker during the early stages of amyloid angiopathy, and the diameter of the lumen decreases, whereas during advanced stages, the walls become thinner and the lumen becomes larger. In addition, we assessed the overall severity of amyloid angiopathy. This showed that thinner arterial walls and the severity of amyloid angiopathy were correlated to dementia. In a multivariate model that integrates the other macroscopic and microscopic lesions that may be implied in the mechanism of cognitive impairment, the severity of amyloid angiopathy per se explained 10% of the variability in the cognitive impairment. PMID- 12898154 TI - Human ependymomas reveal frequent deletions on chromosomes 6 and 9. AB - Ependymomas are glial tumors of the brain and spinal cord. Genetic aberrations associated with the development of these tumors have not been fully identified yet. In previous cytogenetic and comparative genomic hybridization studies, multiple genomic imbalances in ependymomas were found, including partial or whole chromosome losses (1p, 4q, 6q, 9, 10, 11, 13, 16, 17, 19q, 20q and 22q). The aim of this study was to map particularly the commonly affected regions in ependymomas. Thirty-three pairs of matched normal and tumor specimens from ependymoma patients were genotyped using 34 polymorphic microsatellite markers distributed over 15 chromosomes. Loss of heterozygosity (LOH) was found in 26 of 33 tumors (78.8%). The most frequent LOHs were found on the long arms of chromosomes 6 (30.3%) and 9 (27.3%). LOH was also detected on 3p14 (13.3%), 10q23 (10.3%) and 11q (18.2%). Because of the high percentage of LOH on chromosome 6 and 9, we conclude that important tumor suppressor genes are situated on these two chromosomes. PMID- 12898155 TI - Expression of apolipoprotein E in ballooned neurons-comparative immunohistochemical study on neurodegenerative disorders and infarction. AB - Apolipoprotein E (ApoE) in neurons is suggested to play crucial roles in neuronal degeneration and regeneration. We used antibodies against ApoE and phosphorylated neurofilament (pNF) to investigate the immunohistochemical features of ballooned neurons (BNs) in infarction and in various chronic degenerative disorders, including Pick body disease, corticobasal degeneration/progressive supranuclear palsy, Alzheimer's disease, and frontotemporal dementia. BNs in these chronic degenerative processes were intensely labeled with the anti-pNF as reported, whereas BNs in infarction showed less intense pNF-like immunoreactivity (IR). In addition, BNs in infarction were characterized by an intense ApoE-like IR. This ApoE-like IR was inconsistent or less intense in BNs in the chronic degenerative processes. The rarity of ApoE-positive glial cells in the vicinity of ApoE positive BNs suggests that accumulated ApoE in BNs is generated in the neurons. Accumulation of ApoE in BNs in infarction may be linked to a regenerative process after acute transection of axons, which seems compromised in chronic degenerative processes. PMID- 12898156 TI - Neuropathological analysis in spinal muscular atrophy type II. AB - We performed a neuropathological analysis, including in situ nick end labeling (ISEL) and immunohistochemistry, of two cases of clinicogenetically confirmed infantile spinal muscular atrophy (SMA) type II. Both cases showed severe reduction of the motor neurons and gliosis in the spinal cord and brain stem, although the occurrences of central chromatolysis and ballooned neurons were not frequent. Clark's and lateral thalamic nuclei, which are usually altered in SMA type I, were spared, whereas Betz cells in the precentral gyrus and large myelinated fibers in the lateral funiculus were reduced in number. Regarding apoptosis, only the younger case demonstrated a few ISEL-positive nuclei in the dorsal horn, with reduced Bcl-x expression level in the Purkinje cells. Unlike SMA type I, the expression of neurofilaments was not disturbed and the reduction in synaptophysin expression level in the anterior horn was mild. An oxidative stress-related product was deposited in atrophic motor neurons in the spinal cord, and neurons with nuclei immunoreactive for 8-hydroxy-2'-deoxyguanosine were found in the lateral thalamus. In contrast, the expression of glial glutamate transporters was not altered. These data suggest that oxidative stress and, to a lesser extent, apoptotic cell death, but not disturbed neurofilament metabolism or excitotoxicity, may be involved in neurodegeneration in SMA type II. PMID- 12898157 TI - Distal effects in a model of proximal axonopathy: 3,3'-iminodipropionitrile causes specific loss of neurofilaments in rat vestibular afferent endings. AB - 3,3'-Iminodipropionitrile (IDPN) is a neurotoxic compound that causes both a proximal neurofilamentous axonopathy and loss of the vestibular sensory hair cells. We used immunocytochemistry to examine changes in the expression of heavy, medium and light neurofilament (NF-H, NF-M, NF-L) proteins in the afferent terminals of vestibular sensory epithelia after IDPN exposure in rats. Acute, repeated and subchronic IDPN exposure induced a marked loss of NFs in the nerve terminals. The effect of subchronic IDPN was specific, as demonstrated by comparison with the synaptic membrane protein SNAP-25. In addition, Western blot analysis indicated specific loss of NFs in the vestibular receptors. Ultrastructural analysis revealed that afferent endings in the vestibular receptors were significantly preserved in animals exposed to subchronic IDPN, but that these endings showed NF segregation from microtubules followed by NF loss. These effects were closely paralleled by ultrastructural changes in the nerve terminals, particularly in the afferent contacts with the hair cells, and preceded hair cell loss. Thus, distal NF loss and nerve terminal pathology occur in the IDPN model of proximal neurofilamentous axonopathy. Similar distal pathology could also occur in human diseases characterized by proximal axonal swellings, particularly in amyotrophic lateral sclerosis. PMID- 12898158 TI - Absence of detectable alterations in the putative tumor suppressor gene BTRC in cerebellar medulloblastomas and cutaneous basal cell carcinomas. AB - The sonic hedgehog signal transduction pathway is aberrantly activated in the majority of cutaneous basal cell carcinomas and a subset of cerebellar medulloblastomas. The latter tumors may also show activation of the wingless (Wnt) signaling pathway. In Drosophila, the F-box/WD40-repeat containing protein Slimb has been shown to function as an intracellular negative regulator of both pathways. The BTRC gene (beta-transducin repeat-containing protein) is a human homolog of Slimb that is located at 10q24.3, a chromosome region frequently deleted in medulloblastomas. Here, we report on the mutational analysis of BTRC in 91 human tumors, including 66 primitive neuroectodermal tumors (PNETs) of the central nervous system (62 medulloblastomas and 4 supratentorial PNETs) and 25 cutaneous basal cell carcinomas (BCCs). These analyses revealed no tumor associated BTRC mutations. BTRC transcripts were expressed in non-neoplastic brain tissue, normal skin, as well as in all PNETs and BCCs analyzed by real-time reverse transcription-PCR. Two novel BTRC transcript variants were expressed at higher levels in non-neoplastic brain tissue than in normal skin and the investigated tumors. Taken together, our results indicate that PNETs and BCCs neither show mutations nor loss of mRNA expression of BTRC. Therefore, BTRC alteration does not appear to be involved in the pathogenesis of these neoplasms. PMID- 12898159 TI - Sensitivity and specificity of epithelial membrane antigen staining patterns in ependymomas. AB - Pattern and extent of epithelial membrane antigen (EMA) immunoreactivity in ependymomas as compared to other glial tumors have only been investigated in small series. To determine sensitivity and specificity of EMA staining, 54 ependymomas were evaluated in comparison to 54 glioblastomas, 43 fibrillary astrocytomas and 21 oligodendrogliomas. Distinct punctate intracytoplasmic EMA immunoreactivity was observed in 48/54 ependymomas (89%), whereas ring-like EMA staining was observed in 17/54 ependymomas (31%). Apart from the absence in most myxopapillary ependymomas, neither staining pattern was related to tumor grade or localization. Dot-like EMA immunoreactivity was less frequently observed in glioblastomas [32/54 (59%), P<0.05 vs ependymomas], fibrillary astrocytomas [10/43 (23%), P<0.001 vs ependymomas] and oligodendrogliomas [2/21 (10%), P<0.001 vs ependymomas], whereas ring-like EMA staining was absent. Sensitivity and specificity of punctate EMA staining for the diagnosis of ependymoma as compared to other glial tumors were determined: A finding of 5 EMA dots/high-power field was associated with a sensitivity of 72% and a specificity of 81%. The presence of ring-like EMA positive structures was less sensitive (32%), but highly specific (100%). To conclude, distinct punctate and ring-like EMA staining might serve as sensitive and specific markers of ependymal differentiation in glial tumors and, thus, may aid the diagnosis of ependymoma. PMID- 12898160 TI - Prenatal vitamin E improves lung and heart hypoplasia in experimental diaphragmatic [correction of diaphragamatic] hernia. AB - Nitrofen induces in rats diaphragmatic hernia (CDH) with heart and lung hypoplasia by a mechanism involving oxidation. The aim of this study was to examine if prenatal administration of the anti-oxidant agent vitamin E (VitE) prevents to some extent heart and lung hypoplasia. Pregnant rats received on E9.5 either 100 mg of nitrofen alone or followed by 150 IU of VitE on E16.5-E20.5. Control animals received either vehicle or VitE alone. The fetuses were recovered on E21. The hearts and lungs were weighed and DNA and proteins were measured. Sections of the heart and lung were immunohistochemically stained for ki-67, Tunel and TTF-1, and the proportions of proliferating, apoptotic and TTF-1 expressing cells were determined. Cultured human pneumocytes were exposed to the same agents and similarly processed. TTF-1 expression and the proportion of proliferating cells were quantitated. The ANOVA or Kruskall-Wallis tests were used for comparison with p<0.05 as threshold of significance. Nitrofen-exposed rats had decreased lung and heart weight/body weight ratios, lung and heart DNA and protein, lung TTF-1 expression and proportion of proliferating cells in lung and heart. Additional treatment with VitE ameliorated these decreases except for lung TTF-1 and heart weight. In cultured pneumocytes, TTF-1 expression was decreased by nitrofen and rescued by VitE. Cell proliferation followed the same pattern. Antioxidant VitE partially reverses the effects of nitrofen on the heart and lungs of exposed rats. The same effects are observed in cultured human pneumocytes. These results further substantiate the oxidative nature of the effects of nitrofen and suggest that anti-oxidant agents could have a potential clinical application. PMID- 12898161 TI - Abnormal vasculature in intestinal neuronal dysplasia. AB - Intestinal neuronal dysplasia (IND) is an intestinal motility disorder, which clinically resembles Hirschsprung's disease (HD). Adventitial fibromuscular dysplasia (AFMD) consists of proliferation of smooth muscle cells and collagen fibers in the adventitia of blood vessels. The purpose of this study was to investigate vascular abnormalities in large bowel biopsies from patients with isolated HD, IND associated with HD and isolated IND. Large bowel biopsies from patients presenting with isolated HD ( n=23), IND associated with HD ( n=11), isolated IND ( n=16) and normal bowel as controls ( n=6) were investigated using acetylcholinesterase (AChE) histochemistry, van Gieson staining and alpha-smooth muscle actin (alpha-SMA) immunohistochemistry. Increased AChE activity around submucosal vessels was found in 9/16 (56%) cases with isolated IND, 3/11 (27%) cases of IND associated with HD, 5/23 (21%) isolated HD cases and 0/6 controls. AFMD was found in 10/16 (62%) of the isolated IND cases, 4/11 (362) of the cases with IND associated with HD and 4/23 (17%) cases of HD without IND using van Gieson staining. None of the control specimens revealed AFMD. Increased alpha-SMA immunoreactivity filaments were demonstrated in the submucosal vessel wall in 9/16 (56%) of isolated IND and 2/11(18%) of IND associated with HD cases. Normal alpha-SMA immunoreactivity around submucosal vessels was seen in isolated HD and controls. Abnormal submucosal vasculature is a common histological finding in isolated IND and IND associated with HD and may be a useful additional diagnostic feature in these patients. PMID- 12898163 TI - Partially de-epithelialized preputial flap (triangular soft tissue flap): an aid to prevent coronal urethrocutaneous fistulae. AB - A technique is described to fashion a partially de-epithelialized preputial flap, also called triangular soft tissue flap (TSTF), to cover the neourethra in the region of glans, corona and subcorona. This highly vascular TSTF acted as a mechanical barrier against coronal fistula formation in 16 subcoronal hypospadiacs aged 3-12 years who underwent Snodgrass tubularized incised plate urethroplasty. None had complications at 1-year of follow-up. PMID- 12898162 TI - Effects of retinoic acid on the neural crest-controlled organs of fetal rats. AB - Prenatal exposure of rat embryos to retinoic acid induces severe malformations involving various organs. The mechanisms of this embryopathy are known only in part. This study describes the malformations of the neural crest-derived organs in this model and shows that many of them fit into the pattern of disturbed neural crest organogenic control. Pregnant rats were exposed to either all-trans retinoic acid (125 mg/kg; n=17) or vehicle ( n=10) on E10. Fetuses were recovered on E21 and external and internal malformations were sought. The craniofacial area, the trachea, parathyroids, thymus, thyroid, heart, great vessels, and adrenals were examined. In contrast with normal controls, 100% of retinoic acid animals had craniofacial, 94% anorectal, 90% limb, and 55% neural tube defects. The thymus was absent or ectopic in 76%, the parathyroids were absent or single in 88%, and the thyroid was abnormal in 41%. There were neural crest-type (outflow tract and/or pharyngeal aortic arch defects) cardiovascular malformations in 90% and the adrenals were absent in 52%. Interestingly, 9 of 11 (88%) animals with neural tube defects had absent adrenal glands. This association was significant ( p<0.01) by Fisher exact test. Among the complex mechanisms of retinoic acid teratogenesis, severe disturbances of the neural crest pathway play a leading role. The simultaneous development of neural tube defects and adrenal agenesis suggests common pathogenic pathways. PMID- 12898164 TI - Functional capacities of marsupial hearts: size and mitochondrial parameters indicate higher aerobic capabilities than generally seen in placental mammals. AB - This study of marsupial hearts explored the aerobic capacities of this group of mammals; recent information suggests that marsupials possess higher aerobic abilities than previously accepted. Characteristics such as heart mass, mitochondrial features and capillary parameters were examined. A comprehensive study of the heart of red kangaroos was included because of the high maximum oxygen consumption of this species. Goats were also included as a reference placental mammal. Marsupials have a heart that is generally larger than that of placentals. The allometric equation for the relationship between heart mass and body mass for marsupials was M(h)=7.5M(b)(0.944) (M(h) in g and M(b) in kg); the equivalent equation for placental mammals was M(h)=6.0M(b)(0.97). Mitochondrial volume density and inner mitochondrial surface density do not differ between the two mammal groups; although capillary parameters indicated a lower capillary volume in marsupials. Heart size appears to be the major difference between the two groups. The overall pattern seen in marsupials is similar to that of "athletic" placentals and indicates a relatively high aerobic potential. PMID- 12898165 TI - Cell volume affects glycogen phosphorylase activity in fish hepatocytes. AB - The activity of glycogen phosphorylase (GPase) in the active a-form (GPase a) is dependent on the hydration state of hepatocytes. We establish that GPase a catalysis in catfish (Ameiurus nebulosus) hepatocytes is a function of medium osmolarity and that a linear relationship exists between GPase a activity and osmolarity between 254 mosmol l(-1) and 478 mosmol l(-1). Exposure of isolated hepatocytes to hyperosmotic media increases enzyme activity up to 7-fold, indicative of covalent phosphorylation. GPase activation associated with cell shrinkage peaks within 10 min of exposure. The average degree of activation (2.7 fold-increase of GPase a) is only slightly less than in hepatocytes exposed to glucagon (3.1-fold-increase) under isosmotic conditions; with glucagon, the maximum is reached within 2 min. Phosphorylation status remains elevated during the entire 40 min experimental period; cells do not undergo regulatory volume increase (RVI) during this period and do not regain pre-exposure volume. We interpret the increased GPase a activity as an inherent response to hyperosmotic stress, likely brought about by molecular crowding. Activation of the enzyme results in increased glucose production from endogenous glycogen. Glucose is not retained in the liver cells, but may act as an oxidative substrate in extrahepatic tissues for the increased metabolic demand of ion regulation. Protein kinase A or intracellular Ca(2+) make apparently small contributions to the activation of GPase, leaving us to speculate on alternate routes of enzyme activation. Conversely, hepatocyte swelling in hyposmotic medium leads to significant decreases in GPase a activity and curtailed glucose output. A minimum is attained in 10 min, and pre-insult rates are re-established within 40 min, somewhat lagging behind readjustment in cell volume by regulatory volume decrease (RVD). We conclude that cell swelling and subsequent RVD do not signify stress to the cells and metabolic demand may be decreased under cell swelling conditions. Alteration of GPase phosphorylation with extracellular osmolarity appears to be a general phenomenon, since we also find it in hepatocytes of another freshwater catfish (Clarias batrachus) and a marine scorpaenid (Sebastes caurinus). PMID- 12898166 TI - Freeze or dehydrate: only two options for the survival of subzero temperatures in the arctic enchytraeid Fridericia ratzeli. AB - Hygrophilic soil animals, like enchytraeids, overwintering in frozen soil are unlikely to base their cold tolerance on supercooling of body fluids. It seems more likely that they will either freeze due to inoculative freezing, or dehydrate and adjust their body fluid melting point to ambient temperature as has been shown for earthworm cocoons and Collembola. In the present study we tested this hypothesis by exposing field-collected adult Fridericia ratzeli from Disko, West Greenland, to freezing temperatures under various moisture regimes. When cooled at -1 degrees C min(-1) under dry conditions F. ratzeli had a mean temperature of crystallisation ( T(c)) of -5.8 degrees C. However, when exposed to temperatures above standard T(c) for 22 h, at -4 degrees C, most individuals (90%, n= 30) remained unfrozen. Slow cooling from -1 degrees C to -6 degrees C in vials where the air was in equilibrium with the vapour pressure of ice resulted in freezing in about 65% of the individuals. These individuals maintained a normal body water content of 2.7-3.0 mg mg(-1) dry weight and had body fluid melting points of about -0.5 degrees C with little or no change due to freezing. About 35% of the individuals dehydrated drastically to below 1.1 mg mg(-1) dry weight at -6 degrees C, and consequently had lowered their body fluid melting point to ca. -6 degrees C at this time. Survival was high in both frozen and dehydrated animals at -6 degrees C, about 60%. Approximately 25% of the animals (both frozen and dehydrated individuals) had elevated glucose concentrations, but the mean glucose concentration was not increased to any great extent in any group due to cold exposure. The desiccating potential of ice was simulated using aqueous NaCl solutions at 0 degrees C. Water loss and survival in this experiment were in good agreement with results from freezing experiments. The influence of soil moisture on survival and tendency to dehydrate was also evaluated. However, soil moisture ranging between 0.74 g g(-1) and 1.15 g g(-1) dry soil did not result in any significant differences in survival or frequency of dehydrated animals even though the apparent wetness and structure of the soil was clearly different in these moisture contents. PMID- 12898167 TI - The effect of complexity on the discrimination of oriented bars by the honeybee (Apis mellifera). AB - Visual discrimination of black bars by honeybees was studied in a Y-choice apparatus with fixed vertical patterns at constant range. The problem is to discover how bees remember different degrees of complexity of the orientation cue. Previous conclusions with parallel gratings and single bars disagree. With broad bars versus orthogonal bars, the bees learn the orientation cue if the bars are centred at the same place, but they learn the position cue in the vertical direction when the bars are at different places on the two targets. With several bars on each target, the bees learn their orientation and positions. As fixed patterns increase in complexity, the bees follow a simple rule, to look only at the range of places where the cues were displayed. The frame of reference is disrupted when a black spot is added to the training pattern. There is abundant evidence that the bees do not re-assemble the pattern or learn shapes. The filters that detect the position and orientation cues are coarsely tuned, so that they respond in a graded way, but the memory of the range of directions of the cue, as seen from the point of choice, is more exact. PMID- 12898168 TI - Gradual frequency rises in interacting black ghost knifefish, Apteronotus albifrons. AB - The present paper highlights the relationship between social status and production of gradual frequency rises in interacting Apteronotus albifrons. The gradual frequency rise production was mathematically inferred and a discrete classification deliberately avoided. The results showed little gradual frequency rise production before the hierarchy settlement. Afterwards, only the dominant fish kept this gradual frequency rise production at low levels, while the subdominant fish drastically increased it in all following interaction contexts. The hypothesis of gradual frequency rises being involved in communication as submissive signals was thus strengthened. PMID- 12898169 TI - Wasp uses venom cocktail to manipulate the behavior of its cockroach prey. AB - The sting of the parasitoid wasp Ampulex compressa is unusual, as it induces a transient paralysis of the front legs followed by grooming behavior and then by a long-term hypokinesia of its cockroach prey. Because the wasp's goal is to provide a living meal for its newborn larva, the behavioral changes in the prey are brought about by manipulating the host behavior in a way beneficial to the wasp and its offspring. To this end, the wasp injects its venom cocktail with two consecutive stings directly into the host's central nervous system. The first sting in the thorax causes a transient front leg paralysis lasting a few minutes. This paralysis is due to the presence of a venom component that induces a postsynaptic block of central cholinergic synaptic transmission. Following the head sting, dopamine identified in the venom appears to induce 30 min of intense grooming. During the long-term hypokinesia that follows the grooming, specific behaviors of the prey are inhibited while others are unaffected. We propose that the venom represses the activity of head ganglia neurons thereby removing the descending excitatory drive to the thoracic neurons. PMID- 12898170 TI - Role of positive urethrovesical feedback in vesical evacuation. The concept of a second micturition reflex: the urethrovesical reflex. AB - Upon feeling the urge to urinate, the urinary bladder contracts, the urethral sphincters relax and urine flows through the urethra. These actions are mediated by the micturition reflex. We investigated the hypothesis that vesical contraction is maintained by positive feedback through continuous flow of urine through the urethra, and that the cessation of urine flow aborts detrusor contraction. Normal saline was infused into the urinary bladders of 17 healthy volunteers (age 35.2 years+/-4.2(SD); ten women and seven men) at a rate of 100 ml/min. On urge, which occurred at a mean volume of 408.6 ml+/-28.7 of saline, the subject micturated while the vesical and urethral pressures during voiding were being recorded; residual urine was measured. The test was repeated after anesthetizing the urethra with xylocaine gel or, on another occasion, after applying a bland gel. On micturition, the urine was evacuated as a continuous stream without straining; no residual fluid was collected. After urethral anesthetization, the fluid came out of the urethra in multiple intermittent spurts and only with excessive straining. There was a large amount of residual fluid (184.6 ml+/-28.4). The results of bland gel application showed no significant difference ( P>0.05) from those without gel. Detrusor contraction during micturition is suggested to be maintained by positive urethrovesical feedback elicited by the continued passage of urine through the urethra. This feedback seems to be effected through the urethrovesical reflex, which produces vesical contraction on stimulation of the urethral stretch receptors. Abortion of this reflex by urethral anesthetization resulted in failure of detrusor contraction and excessive straining was needed to achieve bladder evacuation in multiple spurts. The urethrovesical reflex is thus assumed to constitute a second micturition reflex responsible for the continuation of detrusor contraction and urination. The role of this reflex in the pathogenesis of micturition disorders needs to be studied. PMID- 12898171 TI - Apical dissection during radical retropubic prostatectomy without ligature. AB - During radical retropubic prostatectomy, hemostasis of the venous vascular plexus is of the utmost importance for avoiding excessive blood loss and ensuring optimal preservation of urethral length, complete extirpation of all apical prostatic notches and careful preservation of the parasympathetic nerves ('neurovascular bundles'). We have developed a careful bipolar coagulation of the venous plexus that results in a significant reduction of intra-operative blood loss, and the consequent need for transfusion, compared to the standard approach. Furthermore, this coagulation approach results in improved visibility, allowing maximum preservation of urethral length, complete extirpation of all apical prostatic notches and improved application of the nerve sparing technique compared to the standard approach. Follow-up after a mean of 14 months suggests that postoperative continence and potency is at least comparable to the standard approach. However, long-term follow-up and multicenter studies will show if this modification results in equally effective cancer control and equivalent functional results compared to the standard approach. PMID- 12898172 TI - A simple and reliable continent bladder stoma constructed from bladder wall. AB - Patients presenting with recurrent bladder neck strictures or severe incontinence refractory to all forms of infravesical treatment usually require chronic catheterization or supravesical urinary diversion. As an alternative, we use a simple continent bladder stoma made from the bladder wall. After extraperitoneal bladder mobilization, a Lapides-like 4 x 8 cm U-shaped bladder flap is excised and sutured to a tube. Depending on the location of the stoma, the base of the flap can be chosen and/or the length of the flap can be adapted. After bladder neck closure the tube is reflected and, after incision and dissection of the detrusor in the middle of the base of the flap, embedded in Lich-Gregoir's technique. The distal end of the tube is then sutured to the stoma site. Five of our patients now have a minimum follow-up of 11 months (mean 17 months). All are continent with a bladder capacity between 250 and 560 ml. In two, stomal stenosis at the skin level required epifascial stomal revision. PMID- 12898173 TI - Evolution of endoscopic extraperitoneal radical prostatectomy (EERPE)--technical improvements and development of a nerve-sparing, potency-preserving approach. AB - During the last decade, laparoscopy has become a standard technique in the armoury of the urologist due to constant technological advances and refinements. Laparoscopic radical prostatectomy (LRPE), although technically demanding and associated with a considerable learning curve, has become the surgical procedure of choice in selected and specialized urological centers around the globe for patients with clinically localized prostate cancer. However, a major drawback of LRPE is the transperitoneal route of access to the extraperitoneal prostate. The principal disadvantages of LRPE are potential intraperitoneal complications, such as bowel injury, ileus, intraperitoneal bleeding, intraperitoneal urinary leakage, intraperitoneal adhesion formation and concomitant small bowel obstruction. Endoscopic extraperitoneal radical prostatectomy (EERPE) is a further advancement of minimal invasive surgery as it overcomes the limitations of LRPE by the strictly extraperitoneal route of access combining the advantages of minimal invasive surgery with those of an extraperitoneal procedure. Based on our growing experience with this procedure, we have introduced several technical modifications, improvements and refinements including a nerve-sparing, potency preserving approach (nEERPE) in an effort to further improve this minimally invasive procedure. PMID- 12898174 TI - Unenhanced helical computed tomography vs intravenous urography in patients with acute flank pain: accuracy and economic impact in a randomized prospective trial. AB - Unenhanced helical computed tomography (UHCT) has evolved into a well-accepted alternative to intravenous urography (IVU) in patients with acute flank pain and suspected ureterolithiasis. The purpose of our randomized prospective study was to analyse the diagnostic accuracy of UHCT vs IVU in the normal clinical setting with special interest on economic impact, applied radiation dose and time savings in patient management. A total of 122 consecutive patients with acute flank pain suggestive of urolithiasis were randomized for UHCT ( n=59) or IVU ( n=63). Patient management (time, contrast media), costs and radiation dose were analysed. The films were independently interpreted by four radiologists, unaware of previous findings, clinical history and clinical outcome. Alternative diagnoses if present were assessed. Direct costs of UHCT and IVU are nearly identical (310/309 Euro). Indirect costs are much lower for UHCT because it saves examination time and when performed immediately initial abdominal plain film (KUB) and sonography are not necessary. Time delay between access to the emergency room and start of the imaging procedure was 32 h 7 min for UHCT and 36 h 55 min for IVU. The UHCT took an average in-room time of 23 min vs 1 h 21 min for IVU. Mild to moderate adverse reactions for contrast material were seen in 3 (5%) patients. The UHCT was safe, as no contrast material was needed. The mean applied radiation dose was 3.3 mSv for IVU and 6.5 mSv for UHCT. Alternative diagnoses were identified in 4 (7%) UHCT patients and 3 (5%) IVU patients. Sensitivity and specificity of UHCT and IVU was 94.1 and 94.2%, and 85.2 and 90.4%, respectively. In patients with suspected renal colic KUB and US may be the least expensive and most easily accessable modalities; however, if needed and available, UHCT can be considered a better alternative than IVU because it has a higher diagnostic accuracy and a better economic impact since it is more effective, faster, less expensive and less risky than IVU. In addition, it also has the capability of detecting various additional renal and extrarenal pathologies. PMID- 12898175 TI - Pneumonia in neutropenic patients. AB - Radiologists have a special role in the management of neutropenic patients. The appropriate investigational technique, frequently targeted differential diagnosis, and the special needs of these patients, need to be understood. Early detection of a focus is the major goal in febrile neutropenic patients. As pneumonia is the most common focus, chest imaging is a special radiological task. The sensitivity of chest X-ray, especially in supine position, is known to be low; therefore, the very sensitive high-resolution CT (HRCT) became gold standard in neutropenic hosts and will probably be replaced by thin-section multislice CT (MSCT) in the near future. Costs of high-resolution CT are low in comparison to antibiotics. An infiltrate needs to be localised, so that a physician can utilise this information as a guidance for invasive procedures for further microbiological work-up. The radiological characterisation of infiltrates gives a first and rapid hint to differentiate between different sorts of infectious (typical bacterial, atypical bacterial, fungal) and non-infectious aetiologies. Follow-up investigations need careful interpretation according to disease and concomitant treatment. Due to an increased incidence of fungal infiltrates even with appropriate therapy, follow-up of an infiltrate must use further parameters in addition to lesion size. Temporary exclusion of infectious involvement of the lung with high accuracy remains of special interest for clinicians. PMID- 12898176 TI - Invasive breast cancer: correlation of dynamic MR features with prognostic factors. AB - The purpose of this exploratory study was to correlate kinetic and morphologic MR features with histologic prognostic factors in invasive breast cancer. Sixty-one women with invasive breast cancer underwent dynamic contrast-enhanced MR imaging at 1.5 T, using T1-weighted 3D fast low-angle shot technique. The MR characteristics were correlated with classical pathologic prognostic factors (tumor size, histologic type, grade and lymph node status) and immunohistochemically detected biomarkers [c-erbB-2, p53, Ki-67, and estrogen receptor (ER)]. Univariate and multivariate statistical analyses were performed. Presence of rim enhancement pattern, early maximal enhancement and washout phenomenon were independently associated with established predictors of poor prognosis (higher histologic grade, positive Ki-67, and negative ER status). Our results suggest that these MR signs are not only important in differentiating benign from malignant lesions, but may also be useful to noninvasively identify highly aggressive breast carcinomas. PMID- 12898177 TI - Elimination of PPV and PNRSV through thermotherapy and meristem-tip culture in nectarine. AB - The plum pox virus (PPV) and prunus necrotic ringspot virus (PNRSV) cause serious disease problems in stone-fruit trees. In this work, the possibility of obtaining plant material free from these viruses through thermotherapy and meristem-tip culture from infected nectarine shoots (Prunus persica var. nectarina Max, cv. 'Arm King') was studied. In addition, the detection of these viruses in in vitro cultures and young acclimatized plantlets with double antibody sandwich-enzyme linked immunosorbent assay (DAS-ELISA) and multiplex reverse transcriptase polymerase chain reaction (RT-PCR) was studied. Meristem-tip explants (0.8-1.3 mm) derived from sprouted buds of winter wood and spring shoots from field grown plants had a 2-5% regeneration response. However, application of thermotherapy to potted nectarine trees (3 weeks at a maximum temperature of 35 degrees C) facilitated excision of longer meristem tips (1.3-2.0 mm) that resulted in a significantly higher regeneration response (38%) in woody plant medium (WPM) without plant growth regulators. Such explants formed multiple shoots with the addition of 8 microM benzylaminopurine and 0.8 microM indoleacetic acid. When they were tested for the presence of PPV and PNRSV, 86% and 81% were found to be virus-free as detected by DAS-ELISA and multiplex RT-PCR, respectively. Individual shoots excised from virus-free cultures readily rooted in vitro (half strength WPM plus 2 microM indolebutyric acid) and grew to plantlets. The combination of an efficient protocol for virus elimination and the establishment of highly sensitive diagnostics resulted in the production of nectarine plants free from PPV and PNRSV. PMID- 12898178 TI - Regeneration and Agrobacterium-mediated transformation of hop (Humulus lupulus L.). AB - An efficient procedure for direct organogenesis and regeneration of hop (Humulus lupulus L.) was established. For the first time Agrobacterium-mediated genetic transformation of hop (cv. "Tettnanger") was achieved. Shoot internodes from in vitro cultures were identified as the most suitable type of explant for regeneration. Using this type of explant, a shoot-inducing medium was developed that supported direct organogenesis of approximately 50% of the explants. Plantlets were successfully rooted and transferred to the greenhouse. Overall, in less than 6 months hop cultures propagated in vitro were regenerated to plants in the greenhouse. Agrobacterium-mediated genetic transformation was performed with the reporter gene GUS (beta-glucuronidase). The presence and function of transgenes in plants growing in the greenhouse was verified by PCR (polymerase chain reaction) and enzyme assay for GUS activity, respectively. We have obtained 21 transgenic plants from 1,440 explants initially transformed, yielding an overall transformation efficiency of 1.5%. PMID- 12898179 TI - Inhibitory effect of cyclo-oxygenase-2 inhibitor on the production of matrix metalloproteinases in rheumatoid fibroblast-like synoviocytes. AB - Cyclo-oxygenase (COX)-2 has been associated with inflammation in rheumatoid arthritis (RA), but its role in joint destruction remains unclear. In this study, we investigated the effect on cultured rheumatoid fibroblast-like synoviocytes (FLS) of the selective COX-2 inhibitor celecoxib on the expression of matrix metalloproteinases (MMPs), which play an important role in tissue degradation and angiogenesis in rheumatoid synovium. Treatment with nontoxic doses of celecoxib resulted in dose-dependent inhibition of MMP-1, -2, and -3 secretion from FLS when measured by enzyme-linked immunosorbent assay. Celecoxib suppressed proinflammatory cytokines (tumor necrosis factor-alpha and interleukin-1beta) induced augmentation of the gelatinolytic activity on zymography. These results suggest that COX-2 inhibitors might influence matrix degradation or angiogenesis in RA by downregulating the expression of various MMPs in rheumatoid FLS. PMID- 12898181 TI - Import and assembly of Neurospora crassa Tom40 into mitochondria of Trypanosoma brucei in vivo. AB - The TOM complex (translocase of the mitochondrial outer membrane) is a dynamic, multisubunit protein complex. Tom40 is the major component of the complex and forms the preprotein conducting pore. To determine if a heterologous Tom40 could be properly targeted and assembled into the Trypanosoma brucei mitochondrial outer membrane, an ectopic copy of a gene encoding Neurospora crassa Tom40 (NcTom40) was expressed in procyclic trypanosomes from a tetracycline regulated procyclic acidic repetitive protein promoter. The level of NcTom40 expression was found to be maximal within 20-26 h of induction with tetracycline. Immunoblot analysis of subcellular fractions showed that NcTom40 was enriched in the mitochondrial fraction. Alkali extraction of isolated mitochondria revealed that NcTom40 was assembled as an integral membrane protein and limited proteolysis demonstrated that it was present in the outer membrane of the mitochondria. These data demonstrate that a heterologous mitochondrial protein containing internal targeting information can be correctly targeted to T. brucei mitochondria. Following blue native gel electrophoresis, the NcTom40 protein was found in a 370 kDa complex which may contain T. brucei Tom components. A 16 kDa protein was coimmunoprecipitated from T. brucei mitochondria containing NcTom40 using antisera developed against the N. crassa protein. The 16 kDa protein may represent a component of the T. brucei TOM complex that associates with NcTom40. PMID- 12898182 TI - Bioavailability and pharmacokinetics of the cardioprotecting flavonoid 7 monohydroxyethylrutoside in mice. AB - PURPOSE: The pharmacokinetics and bioavailability of monoHER, a promising protector against doxorubicin-induced cardiotoxicity, were determined after different routes of administration. METHODS: Mice were treated with 500 mg.kg(-1) monoHER intraperitoneally (i.p.), subcutaneously (s.c.) or intravenously (i.v.) or with 1000 mg.kg(-1) orally. Heart tissue and plasma were collected 24 h after administration. In addition liver and kidney tissues were collected after s.c. administration. The levels of monoHER were measured by HPLC with electrochemical detection. RESULTS: After i.v. administration the AUC(0-120 min) values of monoHER in plasma and heart tissue were 20.5+/-5.3 micromol.min.ml(-1) and 4.9+/ 1.3 micromol.min.g(-1) wet tissue, respectively. After i.p. administration, a mean peak plasma concentration of about 130 microM monoHER was maintained from 5 to 15 min after administration. The AUC(0-120 min) values of monoHER were 6.1+/ 1.1 micromol.min.ml(-1) and 1.6+/-0.4 micromol.min.g(-1) wet tissue in plasma and heart tissue, respectively. After s.c. administration, monoHER levels in plasma reached a maximum (about 230 microM) between 10 and 20 min after administration. The AUC(0-120 min) values of monoHER in plasma, heart, liver and kidney tissues were 8.0+/-0.6 micromol.min.ml(-1), 2.0+/-0.1, 22.4+/-2.0 and 20.5+/-5.7 micromol.min.g(-1), respectively. The i.p. and s.c. bioavailabilities were about 30% and 40%, respectively. After oral administration, monoHER could not be detected in plasma, indicating that monoHER had a very poor oral bioavailability. CONCLUSIONS: MonoHER was amply taken up by the drug elimination organs liver and kidney and less by the target organ heart. Under cardioprotective conditions (500 mg/kg, i.p.), the Cmax was 131 microM and the AUC(infinity) was 6.3 microM.min. These values will be considered endpoints for the clinical phase I study of monoHER. PMID- 12898180 TI - Mitochondrial genome diversity: evolution of the molecular architecture and replication strategy. AB - Mitochondrial genomes in organisms from diverse phylogenetic groups vary in both size and molecular form. Although the types of mitochondrial genome appear very dissimilar, several lines of evidence argue that they do not differ radically. This would imply that interconversion between different types of mitochondrial genome might have occurred via relatively simple mechanisms. We exemplify this scenario on patterns accompanying evolution of mitochondrial telomeres. We propose that mitochondrial telomeres are derived from mobile elements (transposons or plasmids) that invaded mitochondria, integrated into circular or polydisperse linear mitochondrial DNAs (mtDNAs) and subsequently enabled precise resolution of the linear genophore. Simply, the selfish elements generated a problem - how to maintain the ends of a linear DNA - and, at the same time, made themselves essential by providing its solution. This scenario implies that insertion or deletion of such resolution elements may represent relatively simple routes for interconversion between different forms of the mitochondrial genome. PMID- 12898183 TI - Complete regression of primary gastric plasmacytoma following Helicobacter pylori eradication. AB - We describe the first case of a primary gastric plasmacytoma stage I completely regressed following Helicobacter pylori (H.pylori) eradication. The patient, a 61 year-old man, had a long history of chronic gastritis and gastric ulcers with recurrent gastrointestinal hemorrhage. Diagnosis of H.pylori infection was based on the positive urease breath test, the elevated titers of serum anti- H.pylori antibodies, and the detection of the bacterium in gastric mucosa biopsy specimens. Diagnosis of gastric plasmacytoma was based on the findings of histopathology, immunocytochemistry and in situ hybridization. Eradication of H.pylori with antibiotics was followed by disappearance of endoscopic and histopathologic features of the gastric tumor 3 months after the completion of the treatment. No relapse has been documented 20 months after the initial diagnosis of plasmacytoma. A possible causal relationship between the tumor and the underlying H.pylori infection is discussed. PMID- 12898184 TI - CD20-negative DLBCL transformation after rituximab treatment in follicular lymphoma: a new case report and review of the literature. AB - Rituximab is a monoclonal antibody against the CD20 molecule which is used to treat B-cell lymphomas. In 60% of low-grade B lymphomas in which rituximab was effective at first, there was no clinical response in a second treatment and a few cases of follicular lymphomas (FL) with transformation to diffuse large B cell lymphoma (DLBCL) have been reported. We describe a new case and hypothesize about the mechanisms of transformation: a 52-year-old man, in follow-up during 8 years for FL, who after rituximab treatment and complete remission of FL showed progressive disease involving the liver and duodenal mucosa. Immunohistochemical and molecular studies were performed on paraffin-embedded tissue samples of lymph nodes, the small intestine, and liver tumors. After rituximab treatment, biopsies of a liver lesion and the small bowel both showed CD20-negative large B-cell lymphoma. Molecular study of the initial and relapse specimens shows a CDR2 IgH rearrangement with the same height and t14;18 (MBR). The rapid relapse with the same rearrangement of IgH seems to support the interpretation that the change of grade of lymphoma and loss of CD20 expression occurred before rituximab treatment. The existence of a varying proportion of a CD20-negative cell population in every B-cell lymphoma which does not respond to rituximab should therefore be considered. The reduction of CD20 expression could be a resistance mechanism to rituximab retreatment in DLBCL as a consequence of the progression of low-grade B-cell non-Hodgkin's lymphoma (B-NHL). It is necessary to perform new biopsies to evaluate CD20 expression in relapse or the progression of B-cell lymphoma after rituximab treatment. PMID- 12898185 TI - Reduction of leukocyte count is associated with thalidomide response in treatment of multiple myeloma. AB - Fifty Taiwanese patients with relapsed and/or refractory multiple myeloma (MM) were treated with thalidomide on a dose-escalation schedule, commencing with 100 mg/d nightly and incremented either to the maximally tolerated dose or 800 mg/d. Twenty-two patients (44%) responded, with 10 (45.5%) classified as partial remission and 12 (54.5%) minimal response (MR). Complete response did not occur. Of the 28 non-responders, 14 were progressive disease and 14 stable. The median time from commencement of thalidomide treatment to initial achievement of MR was 29 days (range, 8~155), and the corresponding thalidomide dose was 200 mg/d (range, 100~500). The median tolerated dose of thalidomide for the entire sample was 400 mg/d (range, 100~800), with only two (4%) able to tolerate 800 mg/d. Comparing responsive and non-responsive patients, statistically significant differences were not demonstrated for any characteristics except for CRP level and percentage cytogenetic change, which was slightly higher in the latter group relative to the former. Of particular interest, 18 of the 22 responders experienced transient reduction of leukocyte count preceding the attainment of significant reduction in M-proteins in comparison to only four of the 28 non responders (82% vs. 14%; p<0.001). The median time from commencement of thalidomide treatment to attainment of minimal leukocyte count was 28 days (range, 7~150), with a mean of 2.19x10(9)/l (range, 0.96~3.35x10(9)/l). Leukopenia was generally transient, with rapid recovery despite subsequent continuation of thalidomide. Levels of other non-hematologically adverse effects attributed solely to thalidomide were generally acceptable. For 25 patients, thalidomide treatment was supplemented with low-dose dexamethasone (4 mg, every other day). Of these, 11 had relapsed from and 14 were primarily refractory to thalidomide treatment. Nine of the 25 dexamethasone-supplemented patients were responders (36%). Of particular note were the unusual events noted with this thalidomide-dexamethasone combination, including vascular thrombosis, acute cholecystitis, idiopathic interstitial lung disease and sudden cardiac death. Our results suggest that thalidomide is also effective for Taiwanese patients with refractory and/or relapsed MM. Importantly, the transient reduction in leukocyte count after commencement of thalidomide treatment may serve as a clinical predictor for response. Adverse effects should be carefully monitored when combining thalidomide and dexamethasone, however. PMID- 12898186 TI - A case of myelodysplastic syndrome developed blastic crisis of chronic myelogenous leukemia with acquisition of major BCR/ABL. AB - We describe a rare case of myelodysplastic syndrome that developed chronic myelogeneous leukemia with acquisition of Philadelphia chromosome. The major BCR/ABL transcript was confirmed by molecular analysis. Shortly thereafter, the patient showed transformation to blastic crisis. Hematological remission was achieved after 3 months of treatment with imatinib mesylate. The patient relapsed with additional chromosomal abnormalities and the disease became refractory to the treatment. Acquisition of the Philadelphia chromosome is an infrequent event in myelodysplastic syndrome, and the addition of this change to the initial genetic abnormality that caused MDS may have been associated with the accelerated clinical course of this patient. PMID- 12898187 TI - Molecular basis of weak D in Taiwanese. AB - Two genes, RHD and RHCE, encode the antigens of the RH blood group system. The weak D phenotype is caused by many different RHD alleles encoding aberrant RhD proteins, resulting in distinct serologic phenotypes and anti-D immunization. We analyzed seven weak D phenotypes excluding D(el), using polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) and direct sequencing methods to detect the changes of all ten RHD exons. The results show that there are four types of weak D in Taiwanese: one case each for CGG to CAG mutation at codon 10, GTG to ATG mutation at codon 174, and GTG to GAG mutation at codon 270, and four cases for GGT to GAT mutation at codon 282. In conclusion, we present the first data of a molecular basis of weak D in Taiwanese, which suggest a clinically relevant potential for anti-D immunization and may improve transfusion strategy in weak D Taiwanese patients. PMID- 12898188 TI - Growth hormone (GH) deficiency in patients with beta-thalassemia major and the efficacy of recombinant GH treatment. AB - Patients with beta-thalassemia major still suffer growth retardation. After excluding patients with cortisol deficiency, hypothyroidism, hypogonadism, delayed puberty, malnutrition, severe congestive heart failure, and severely impaired liver function, 29 patients were enrolled in this study. Fifteen (52%) patients exhibited growth retardation and underwent two growth hormone (GH) provocation tests. Eight (53%) of the 15 patients had GH deficiency and were subsequently treated with subcutaneous recombinant human GH (Genotropin, Pharmacia Corporation, Sweden). Growth velocity increased from the pretreatment rate of 3.1+/-0.4 cm/year to 7.1+/-1.6 cm/yr (p<0.001) after 1 year and to 6.8+/ 1.3 cm/year (p<0.001) after 2 years. Patients with growth retardation had lower insulin like growth factor-1 (p=0.001) and insulin like growth factor binding protein-3 (p=0.003) levels than those without growth retardation. In patients with beta-thalassemia major, growth retardation is a common complication and GH deficiency plays an important role. Thalassemic patients with GH deficiency can safely increase their growth velocity with recombinant human GH for 2 years; however, the effect on final height still needs to be determined. PMID- 12898189 TI - Bone marrow-derived stem cells and "plasticity". AB - Studies describing plasticity of somatic stem cells have become a focus of interest because clinical applications in the treatment of degenerative diseases would be at hand. In particular, bone marrow-derived cells and their potential to contribute to skeletal and cardiac muscle, liver, neurons and epithelium have recently been studied extensively. Nevertheless, results of these studies have not always been consistent with each other, and yet it remains to be resolved whether plasticity of adult stem cells truly exists. This review will discuss the role of bone marrow-derived stem cells in the field of experimental and clinical plasticity studies. Observations compatible with the concept of stem cell plasticity will be weighed against limitations of the experimental systems employed. PMID- 12898190 TI - A case of multicentric Castleman's disease associated with advanced systemic amyloidosis treated with chemotherapy and anti-CD20 monoclonal antibody. AB - Multicentric Castleman's disease (MCD) is a rare systemic lymphoproliferative disorder with too few patient series reported in the literature to have a clear idea about the etiology, outcome and the best treatment available. Systemic reactive amyloidosis is a very rare complication of MCD and its presence worsens the prognosis. We report a case of a 28-year-old patient with plasma-cell type, human immunodeficiency virus (HIV)-negative and human herpesvirus-8 (HHV-8) negative MCD who responded to treatment with chemotherapy and the anti-CD20 monoclonal antibody, rituximab. Anti-CD20 therapy could be an interesting adjunctive treatment in MCD. PMID- 12898191 TI - FCGR3B gene frequencies and FCGR3 variants in a Chinese population from Zhejiang Province. AB - The human neutrophil antigens HNA-1a, -1b and -1c play an important role in immune neutropenia. The frequencies of the coding FCGR3B genes were determined in different populations. New FCGR3B variants were also found in some populations. This study investigated the FCGR3B gene frequencies and FCGR3 variants in a Chinese population compared with the results of Northern Germans and African Blacks (Uganda). Our results show that the gene frequencies in 413 healthy Chinese individuals from Zhejiang Province were 0.565 for FCGR3B*1, 0.430 for FCGR3B*2 and 0.00 for FCGR3B*3. The genotype frequency of FCGR3B(null) was 0.48% (2/413). Sequencing of FCGR3 revealed that in seven out of 19 Chinese individuals, cloned and sequenced DNA fragments that exhibited variants caused by single nucleotide exchanges at one or more of the polymorphic positions 141, 147, 227, 266 and 277 in exon 3 also existed in this Chinese population. From the present study, it is concluded that the FCGR3B*1 gene is more frequent in a Chinese population from Zhejiang Province than the FCGR3B*2 gene, and the FCGR3B*3 gene seems to be absent, which is in contrast to studies in the white populations. Gene variants caused by single nucleotide exchanges were found in addition to the well-known forms, but the reason for this remains unclear. PMID- 12898192 TI - Sternalis muscle: importance in surgery of the breast. AB - Mastectomy is a commonly performed surgery for carcinoma of breast. During surgery, pectoral fascia is removed and pectoralis major muscle is laid bare. Sternalis is a rare muscle encountered in the subcutaneous plane. We examined the operative records of 1,152 patients who underwent modified radical mastectomies between 1990 and 2000. Patients who underwent conservative breast surgery or radical mastectomy were excluded. Among 1,152 patients who underwent modified radical mastectomy, eight were identified as having sternalis, a subcutaneously placed muscle oriented craniocaudally. The thickness of the muscle varied. The muscle was spared in all patients. Sternalis is a rare muscle in the subcutaneous plane. It should not be mistaken for a mass on mammography. During surgery it is important to be aware of this rare entity and identify the muscle early so that the dissection plane is appropriate. The depth at which internal mammary nodes are irradiated may also vary in the presence of the muscle. In addition, it should not be mistaken for recurrence on follow-up. PMID- 12898193 TI - "Horseshoe kidney" in an ectopic position. A case report. AB - We present a case study of a horseshoe kidney, harvested from a 68-year-old patient who died suddenly from rupture of an aneurysm of the ascending aorta. The kidney was ectopically placed, its isthmus corresponding to the terminal aorta. The two kidneys were unequal in size, the right kidney being larger than the left. We discovered four renal arteries, two on the right and two on the left side, and four renal veins, one left, one middle and two on the right side. The isthmus was blood supplied by the left and the right inferior renal arteries while the venous blood was collected by the left and middle renal veins. PMID- 12898194 TI - The superior epigastric artery does not pass through Larrey's space (trigonum sternocostale). AB - The passage of the superior epigastric artery (SEA), the terminal branch of the internal thoracic artery (ITA), through the inferior orifice of the thorax differs in different reports. According to some, it passes through Larrey's space (trigonum sternocostale), therefore through a diaphragmatic orifice, but according to others it passes in front of the diaphragm and the transverse abdominal muscle. The aim of this study was to determine the position of the SEA in its thoracoabdominal segment. We carried out a series of 14 dissections (10 on embalmed cadavers and 4 on unembalmed cadavers), and a study of images from the Visible Human Project. Dissections always led to the same conclusions. After having dissected the trigonum sternocostale, we observed that no vascular element was present in the space, which was obstructed downwards by the parietal peritoneum and limited forwards by the aponeurosis of the transverse abdominal muscle. Inferior digitations of transversus thoracis were joined with the transversus abdominis. The SEA passed in front of the plane formed by these two muscles while the sternal and costal parts of the diaphragm were behind this plane. Whatever the level of the section of the Visible Human Project, there was always a musculoaponeurotic plane between Larrey's space and the superior epigastric artery and both veins. Larrey's space, or trigonum sternocostale, was limited medially by the lateral border of the sternal part of the diaphragm, laterally by the medial border of the costal part of the diaphragm, and anteriorly by the musculoaponeurotic plane formed by the transversus thoracis above and the transversus abdominis, below without a clear boundary between those muscles. The SEA, the terminal branch of the ITA, passed in front of this musculoaponeurotic plane. PMID- 12898195 TI - The trapezoid form of the trochlea tali. AB - The wedge-shaped superior surface of the trochlea tali may be seen in a proximal view as a trapezium. The angle of this trapezium was calculated by measuring the length, and the smallest and largest widths of the trochlea, and was found to be 16.04 degrees for the left talus and 12.48 degrees for the right talus, respectively. The difference between these angles is highly significant. Due to biomechanical features of the talocrural joint an increase in the angle of the trochlea tali also intensifies the incongruency of this joint in plantar-flexion. Thus, this incongruency is larger on the left side. With this diminution of the joint-surface contact, increased internal rotation or "wobbling" movements are possible. An explanation for these facts might be found in footedness, similar to handedness. When constructing a prosthesis one has to consider that the trochlea tali cannot be exactly mirrored from one side to the other but may have to be calculated separately for each side. The fact that footedness seems to correlate with the angle of the trochlea tali can be also helpful. PMID- 12898196 TI - Persistent left superior vena cava and central venous catheter position: clinical impact illustrated by four cases. AB - Variations in the course of the blood vessels are often incidental findings during clinical examination. A persistent left superior vena cava (LSVC) is really not rare (healthy individuals, 0.3-0.5%; patients with congenital heart disease, 4%) and serious complications have been described during catheterization in adults with LSVC (shock, cardiac arrest, angina). Therefore variations of the superior vena cava should be considered, especially when central venous catheterization via the subclavian or internal jugular vein is difficult. We describe the embryogenesis and the anatomic variations of persistent LSVC. Subsequently we suggest a classification of superior vena cava according to the positioning of a central venous catheter on the chest radiograph: type I, normal anatomy; type II, only persistent left superior vena cava; type IIIa, right and left superior vena cava with connection; type IIIb, right and left superior vena cava without connection. This classification is illustrated by four clinical cases. PMID- 12898198 TI - Total situs inversus: a genetic material bank as a new tool for anatomical research. PMID- 12898199 TI - Periprosthetic fractures in total hip arthroplasty: an epidemiologic study. AB - This study was based on data from the Finnish Arthroplasty Register. From 1990 to 1999, 33,154 primary hip arthroplasties were performed in Finland. Only periprosthetic fractures treated by a revision arthroplasty were registered. The six most used femoral components were compared using survival analysis and Cox's regression model. The incidence of periprosthetic fractures was calculated separately for the years 1990-1994 and 1995-1999. The incidence in the first period was greater than in the latter. Survival analysis and Cox's regression model showed that gender, prosthesis type and age were of no significance as risk factors for periprosthetic fractures. PMID- 12898200 TI - Re: Puolakka et al. Increased volumetric wear of polyethylene liners with more than 3 years of shelf-life time. Int Orthop (2003) 27:153-9. PMID- 12898201 TI - [18F]FLT PET for diagnosis and staging of thoracic tumours. AB - The nucleoside analogue 3'-deoxy-3'-[18F]fluorothymidine (FLT) has been introduced for imaging of tumour cell proliferation by positron emission tomography (PET). This study evaluated the use of FLT in patients with thoracic tumours prior to treatment. Whole-body FLT PET was performed in 16 patients with 18 tumours [17 thoracic tumours (nine non-small cell lung cancers, five oesophageal carcinomas, two sarcomas, one Hodgkin's lymphoma) and one renal carcinoma] before treatment. Fluorine-18 fluorodeoxyglucose (FDG) PET was performed for comparison except in those patients with oesophageal carcinoma. For semi-quantitative analysis, the average and maximum standardised uptake values (avgSUV and maxSUV, respectively) (FLT, 114+/-20 min p.i.; FDG, 87+/-8 min p.i.; 50% isocontour region of interest) was calculated. All 17 thoracic tumours and 19/20 metastases revealed significant FLT accumulation, resulting in easy delineation from surrounding tissue. The additional small grade 1 renal carcinoma was not detected with either FLT or FDG. In most lung tumours (avgSUV 1.5-8.2) and metastases, FLT showed intense uptake. However, one of two spinal bone metastases was missed owing to the high physiological FLT uptake in the surrounding bone marrow. Oesophageal carcinoma primaries (avgSUV 2.7-10.0) and occasional metastases showed particularly favourable tumour/non-tumour contrast. Compared with FDG, tumour uptake of FLT was lower (avgSUV, P=0.0006; maxSUV, P=0.0001), with a significant linear correlation (avgSUV, r2=0.45; maxSUV, r2=0.49) between FLT and FDG. It is concluded that FLT PET accurately visualises thoracic tumour lesions. In the liver and the bone marrow, high physiological FLT uptake hampers detection of metastases. On the other hand, FLT may be favourable for imaging of brain metastases owing to the low physiological uptake. PMID- 12898203 TI - Rare complication of sinus venosus-type atrial septal defect repair. PMID- 12898204 TI - [Tc(CO)(3)](+) chemistry: a promising new concept for SPET? For. PMID- 12898205 TI - [Tc(CO)(3)](+) chemistry: a promising new concept for SPECT? Against. PMID- 12898206 TI - The need for Health Technology Assessments of PET. PMID- 12898208 TI - Expression of GABA transporters, GAT-1 and GAT-3, in the cerebral cortex and thalamus of the rat during postnatal development. AB - The cellular and subcellular localization of two GABA transporters, GAT-1 and GAT 3, was investigated using immunocytochemical methods in the rat cerebral cortex and thalamus during postnatal development. The distribution of the transporters is compared with that of the neuronal marker GABA, and with that of vimentin and of glial fibrillary acidic protein, which identify immature and mature astrocytes, respectively. Our observations show that the two transporters are already expressed at birth in both brain areas with the same cellular localization as in adult rats, as GAT-1 is present in growth cones and terminals only in the cortex, whereas both transporters are expressed in astrocytes in the cortex and thalamus. The distribution of GAT-1 and GAT-3 undergoes postnatal changes reflecting in general the neurogenetic events of the neocortex and thalamus and, more specifically, the maturation of GABAergic innervation. The adult-like pattern of expression is achieved in the third postnatal week in the cortex and in the second postnatal week in the thalamus. The early expression of GAT-1 in GABAergic terminals confirms previous studies showing the existence of neuronal mechanisms of GABA uptake from the embryonic stages. As for the glial localization, the precocious existence of two astrocytic GABA transporters suggests that they operate through different functional mechanisms from birth, whereas their exclusively glial expression in the thalamus indicates that the astroglia plays a major role in the transport, recycling and metabolism of thalamic GABA. PMID- 12898209 TI - Endoreplication and polyploidy in primary culture of rat hepatic stellate cells. AB - Hepatic stellate cells (HSCs), the pericytes of hepatic sinusoids, and liver myofibroblasts (rMFs), cells located in the portal field and around the pericentral area, are the principal fibrogenic cell types of the liver. In cases of liver damage HSCs undergo "activation," i.e., they acquire a myofibroblast like appearance and synthesize huge amounts of extracellular matrix proteins (ECMs). Their proliferation ability, however, is a matter of debate. In fact, during culture the number of rat HSCs decreases, while DNA synthesis activity and DNA content per cell increase (4+/-0.6 times). Together with the decrease in cell number (60+/-19% at day 6 of primary culture compared to day 3), cell volume increases and many HSCs become multinuclear. On the other hand, in cultures of rMFs, cell number increases along with DNA synthesis, and these cells do not become multinuclear. "Activated" HSCs produce higher levels of cyclin D(1) and E(1) transcripts than rMFs, which correlates with their increased levels of phosphorylated retinoblastoma (Rb) protein. In activated HSCs DNA synthesis seems to be associated with polyploidy and increase in cell volume, while DNA synthesis is followed by mitosis in rMFs. PMID- 12898210 TI - Interdependent development of blood vessels and organs. AB - The cardiovascular system is the first functional organ in the vertebrate embryo, and many organs start to develop adjacent to cells of the cardiovascular system. Endothelial cells (EC) form the inner cell lining of blood vessels and represent the major cell type that interacts with developing organs. On the one hand, EC provide organs with signals. These signals determine the location, differentiation and morphology of an organ. On the other hand, EC receive signals from the organ-specific cell types. Such signals give EC organ-specific features that the organ needs to interact with the circulatory system. This review provides the reader with specific examples of an interdependent development of organs and blood vessels. PMID- 12898211 TI - Role of hypoxia in tumor angiogenesis-molecular and cellular angiogenic crosstalk. AB - The mechanisms by which tumors recruit their vasculature has been subject to intense investigations. The acquisition of a functional blood supply seems to be rate-limiting for the ability of a tumor to grow beyond a certain size and to metastasize to other sites. Accumulating evidence indicates that hypoxia and the key transcriptional system, HIF (hypoxia-inducible factor), are the major triggers for new blood vessel growth in malignant tumors. Although vessel growth and maturation are complex and highly coordinated processes requiring the sequential activation of a multitude of factors, there is a consensus that vascular endothelial growth factor and angiopoietin signaling represent crucial steps in tumor angiogenesis. Recent insights into cellular and molecular crosstalk suggest a model in which hypoxia, HIF, and several HIF target genes participate in the coordinated collaboration between tumor, endothelial, inflammatory/hematopoietic, and circulating endothelial precursor cells to enhance and promote tumor vascularization. A well-integrated understanding of this intricate microenvironment may offer new opportunities for therapeutic intervention. PMID- 12898212 TI - A comparative study of the expression of monoamine oxidase-A and -B mRNA and protein in non-CNS human tissues. AB - The distributions of monoamine oxidase (MAO)-A and -B proteins and mRNAs in human heart, lung, liver, duodenum, kidney and vasculature were compared using immunohistochemistry and cRNA in situ hybridisation. MAO-A and -B mRNA were detected in all tissues, to differing extents, but particularly in glomeruli, hepatocytes, and the crypts, muscularis mucosa and muscularis externa of duodenum. Renal proximal and distal tubules and loops of Henle had more intense labelling for mRNA of MAO-B than MAO-A; this was reflected in MAO protein expression. Little immunoreactivity was detected in glomeruli. Hepatocytes expressed MAO-A moderately, but MAO-B strongly. In lungs, similar moderately intense labelling for both MAO mRNAs and immunoreactivities was evident in pneumocytes, and epithelial and smooth muscle cells. Cardiomyocytes contained both MAO isoforms, but with more, albeit moderate, labelling for MAO-A. Both isoforms were expressed equally in duodenal villi, crypts, muscularis externa and mucosa; lower level expression occurred in mucosal and submucosal cells. MAO-A and -B mRNA were detected in endothelia, adventitia and media of a renal interlobular artery, but protein immunoreactivities were chiefly in the adventitia. The data reveal widespread tissue distribution of MAO mRNAs and proteins, but indicate that presence of MAO mRNAs does not invariably reflect quantitatively its protein expression. PMID- 12898213 TI - Deferoxamine reduces and nitric oxide synthase inhibition increases neutrophil mediated myotube injury. AB - We tested the contribution of reactive oxygen species (ROS), reactive nitrogen species (RNS) and the beta 2 integrin CD18 to neutrophil-mediated myotube injury. Human myotubes were cultured with human neutrophils in the presence or absence of inhibitors directed against ROS, RNS, and CD18. Muscle injury was assessed by a (51)Cr release assay. The inclusion of superoxide dismutase (50-500 U/ml) in the culture medium did not affect myotube injury. A significant protective effect was provided by including catalase (600-2400 U/ml), deferoxamine (1-2 mM), or anti CD18 antibody (10 microg/ml) in the culture medium. S-Ethylisothiourea (500-1000 microM), an inhibitor of nitric oxide synthase (NOS), significantly increased myotube injury and reduced nitric oxide (NO) in cultures consisting of only myotubes. In conclusion, neutrophil-mediated skeletal muscle injury appears to be largely dependent on CD18-mediated neutrophil adhesion and iron-dependent hydroxyl radical production. In addition, skeletal muscle NOS activity may protect skeletal muscle against the injury caused by neutrophils. PMID- 12898215 TI - Evaluation of the microsomal glutathione S-transferase 3 (MGST3) locus on 1q23 as a Type 2 diabetes susceptibility gene in Pima Indians. AB - Elevation of plasma glucose concentration may induce generation of oxygen-free radicals, which can play an important role in the progression of diabetes and/or development of its complications. Various glutathione transferases utilize the availability of reduced glutathione for the cellular defense against oxygen-free radicals. One such enzyme is microsomal glutathione S-transferase 3 encoded by MGST3, which maps to chromosome 1q23, a region linked to Type 2 diabetes mellitus (T2DM) in Pima Indians, Caucasian, and Chinese populations. We investigated the MGST3 gene as a potential susceptibility gene for T2DM by screening this locus for single nucleotide polymorphisms (SNPs) in diabetic and non-diabetic Pima Indians. We also measured the skeletal muscle MGST3 mRNA level by Real-Time (RT) PCR and its relationship with insulin action in non-diabetic individuals. We identified 25 diallelic variants, most of which, based on their genotypic concordance, could be divided into three distinct linkage disequilibrium (LD) groups. We genotyped unique representative SNPs in selected diabetic and non diabetic Pima Indians and found no evidence for association with T2DM. Furthermore, inter-individual variation of skeletal muscle MGST3 mRNA was not correlated with differences in insulin action in non-diabetic subjects. We conclude that alterations of MGST3 are unlikely to contribute to T2DM or differences in insulin sensitivity in the Pima Indians. PMID- 12898216 TI - Zaba: a novel miniature transposable element present in genomes of legume plants. AB - A novel family of miniature transposable elements, named Zaba, was identified in pea (Pisum sativum) and subsequently also in other legume species using computer analysis of their DNA sequences. Zaba elements are 141-190 bp long, generate 10 bp target site duplications, and their terminal inverted repeats make up most of the sequence. Zaba elements thus resemble class 3 foldback transposons. The elements are only moderately repetitive in pea (tens to hundreds copies per haploid genome), but they are present in up to thousands of copies in the genomes of several Medicago and Vicia species. More detailed analysis of the elements from pea, including isolation of new sequences from a genomic library, revealed that a fraction of these elements are truncated, and that their last transposition probably did not occur recently. A search for Zaba sequences in EST databases showed that at least some elements are transcribed, most probably due to their association with genic regions. PMID- 12898217 TI - MCB-mediated regulation of cell cycle-specific cdc22+ transcription in fission yeast. AB - The cdc22+ gene of the fission yeast, Schizosaccharomyces pombe, encodes the large subunit of ribonucleotide reductase, and is periodically expressed during the mitotic cell cycle, transcript abundance reaching a maximum at the G1-S boundary. This regulation of expression is controlled by a transcription factor complex called DSC1, which binds to MCB motifs (ACGCGT) present in the promoter of cdc22+. cdc22+ has a complex pattern of MCBs, including two clusters of four motifs each, one of which is located within the transcribed region. We show that both clusters of MCBs contribute to the regulation of cdc22+ expression during the cell cycle, each having a different role. The MCB cluster within the transcribed region has the major role in regulating cdc22+, as its removal results in loss of transcription. The upstream cluster, instead, controls cell cycle-specific transcription through a negative function, as its removal results in expression of cdc22+ throughout the cell cycle. Both MCB clusters bind DSC1. We show that the interaction of DSC1 with the MCB cluster within the transcribed region has a high "on-off" rate, suggesting a mechanism by which DSC1 could activate expression, and still allow RNA polymerase to pass during transcription. Finally, we show that both clusters are orientation-dependent in their function. The significance of these results, in the context of MCB-mediated regulation of G1-S expression in fission yeast, is discussed. PMID- 12898218 TI - Alloplasmic male sterility in Brassica napus (CMS 'Tournefortii-Stiewe') is associated with a special gene arrangement around a novel atp9 gene. AB - To identify regions of the mitochondrial genome potentially involved in the expression of alloplasmic 'Tournefortii-Stiewe' cytoplasmic male sterility (CMS) in Brassica napus, transcripts of 25 mitochondrial genes were analysed in fertile and near isogenic male-sterile plants (BC(8) generation). Differences were detected in the transcription of genes for subunit 9 of ATP synthase (atp9), cytochrome b (cob) and subunit 2 of NADH dehydrogenase (nad2). Structural analysis of these gene regions revealed differences in genome organisation around atp9 between male-sterile and fertile plants. Three atp9 genes, two of which were hitherto unknown, are present in the mitochondria of CMS plants, and rearrangements upstream of one of these genes have generated a chimeric 193-codon ORF, designated orf193. This region is transcribed as a CMS specific bi-cistronic mRNA of 1.58 kb comprising orf193 and atp9. The level of the aberrant 1.58-kb transcript is reduced in plants restored to fertility by as yet uncharacterized nuclear genes. orf193 encodes a polypeptide of 22.7 kDa which exhibits partial sequence identity to the subunit 6 of the ATP synthase complex. However, as it forms an uninterrupted ORF with one of the newly discovered atp9 genes it may also be translated as a chimeric 30.2-kDa protein. It is likely that either or both gene products interfere with the function or assembly of the mitochondrial F(0)F(1)-ATP synthase, thus impairing the highly ATP-dependent process of pollen development. The novel molecular features of alloplasmic 'Tournefortii-Stiewe' CMS are discussed with respect to the other known mechanisms of CMS in B. napus. PMID- 12898220 TI - Genetic analysis of ryanodine receptor function in Caenorhabditis elegans based on unc-68 revertants. AB - The Caenorhabditis elegans ryanodine receptor is encoded by the unc-68 gene, and functions as a Ca2+-induced Ca2+ release channel during muscle contraction. To investigate the factors that suppress calcium release and identify molecules that interact with the ryanodine receptor, we isolated revertants from two unc-68 mutants. Three of the revertants obtained from the null allele unc-68(e540), which displayed normal motility, had intragenic mutations that resulted in failure to splice out intron 21. The other two, kh53 and kh55, had amino acid insertions in the third of the four RyR domains. The brood size and the egg laying rate remain abnormal in these revertants. This suggests the third RyR domain may be required for egg laying and embryogenesis, although we can not determine a molecular mechanism. Five ketamine sensitive revertants recovered from the missense mutant unc-68(kh30) showed altered responses to caffeine, ryanodine, levamisole and ouabain relative to those of the unc-68(kh30) animals. These may carry second-site suppressor mutations, which may define genes for proteins that regulate the Ca2+ concentration in body-wall muscle. One of these mutants, kh52, shows lower motility and higher sensitivity to drugs, and this mutation was mapped to chromosome X. These observations provide a basis for the study of ryanodine receptor functions in embryogenesis and in calcium-mediated regulation of muscle contraction in C. elegans. This is the first study to show that the conserved RyR domain of the receptor acts in egg laying and embryogenesis. PMID- 12898219 TI - Mutation of a conserved CDK site converts a metazoan Elongation Factor 1Bbeta subunit into a replacement for yeast eEF1Balpha. AB - Elongation factor subunit eEF1Bbeta (formerly EF-1beta in plants and EF-1delta in animals) was identified and cloned in a screen for proteins from pea that interact with a cyclin-dependent kinase (CDK). CDKs are enzymes that regulate progression through meiotic and mitotic cell cycles in eukaryotes. eEF1Bbeta and the related protein eEF1Balpha (formerly EF-1beta' in plants and EF-1beta in animals and fungi) can catalyze GTP/GDP exchange on the G-protein eEF1A (formerly EF-1alpha in plants, animals and fungi) during the elongation phase of protein synthesis in eukaryotes. Recombinant Cdc2 and its native homologues from pea extracts associated both in vitro and in vivo with eEF1Bbeta. A Cdc2-cyclin B complex phosphorylated recombinant plant eEF1Bbetas, but not eEF1Balpha. These interactions between CDK and eEF1Bbeta prompted investigations into the in vivo consequences of this relationship. Expression of cDNAs encoding rice or pea eEF1Bbeta subunits failed to complement a Saccharomyces cerevisiae mutant deleted for the eEF1Balpha gene, as was previously observed for the human eEF1Bbeta. However, replacement of Thr91, the sole consensus CDK phosphorylation site in pea eEF1Bbeta, with alanine allowed the pea protein to substitute for eEF1Balpha function in vivo. In addition, this rescued strain was severely cold sensitive, and more sensitive to translational inhibitors than wild-type yeast. Taken together, these results suggest a physiological connection between the cyclin dependent class of kinases and a translational elongation factor in mitotic cells, and provide the first in vivo evidence that an altered form of eEF1Bbeta can serve as the guanine nucleotide exchange factor for eEF1A. PMID- 12898221 TI - Characterization of the Tuber borchii nitrate reductase gene and its role in ectomycorrhizae. AB - The nitrate assimilation pathway represents a useful model system in which to study the contribution of a mycorrhizal fungus to the nitrogen nutrition of its host plant. In the present work we cloned and characterized the nitrate reductase gene (tbnr1) from Tuber borchii. The coding region of tbnr1 is 2,787 nt in length, and it encodes a protein of 929 amino acids. Biochemical and Northern blot analyses revealed that nitrate assimilation in T. borchii is an inducible system that responds mainly to nitrate. Furthermore, we cloned a nitrate reductase cDNA (tpnr1) from Tilia platyphyllos to set up a quantitative real-time PCR assay that would allow us to determine the fungal contribution to nitrate assimilation in ectomycorrhizal tissue. Using this approach we demonstrated that the level of tbnr1 expression in ectomycorhizae is eight times higher than in free-living mycelia, whereas tpnr1 transcription was found to be down-regulated after the establishment of the symbiosis. Enzymatic assays showed that NADPH dependent nitrite formation markedly increases in ectomycorrhizae. These findings imply that the fungal partner plays a fundamental role in nitrate assimilation by ectomycorrhizae. Amino acid determination by HPLC revealed higher levels of glutamate, glutamine and asparagine in symbiotic tissues compared with mycelial controls, thus suggesting that these amino acids may represent the compounds that serve to transfer nitrogen to the host plant. PMID- 12898222 TI - The gyr genes of Salmonella enterica serovar Typhimurium are repressed by the factor for inversion stimulation, Fis. AB - The DNA sequence of the gyr genes from Salmonella enterica serovar Typhimurium revealed strong similarity between gyrB and its counterpart in Escherichia coli. However, the gyrA gene showed similarity to the E. coli homologue only downstream from the Pribnow box of the promoter, with the sequence upstream diverging markedly. Since this region encompasses the binding sites for the Fis DNA binding protein in E. coli, we investigated the possibility that the gyrA genes in the two species might differ in their responses to this regulatory protein. Fis was found to act as a transcriptional repressor of both gyr genes in S. enterica. In electrophoretic mobility shift assays, Fis was found to bind to both the gyrA and gyrB promoters of S. enterica, despite the strong divergence from the E. coli sequence on the part of the former. The binding sites were mapped by DNase I protection assays, and the results are consistent with conservation of the mechanism of Fis-mediated repression between the two bacterial species. PMID- 12898224 TI - Dietary supplementation with Echinacea and development of immunity to challenge infection with coccidia. AB - An experiment was conducted to examine the effects of dietary supplementation with 0.1% and 0.5% ground root preparations of Echinacea purpurea (EP; common name: purple cone flower) on the development of immunity following live vaccination and subsequent challenge with multiple coccidia species in an experimental model using a commercial live vaccine preparation. Effects of immunization and EP supplementation on weight gains before challenge, and weight gains, lesion scores and plasma levels of carotenoids and NO2-+NO3- following challenge were determined. In this experiment, combined live vaccination and feed supplementation with 0.1% or 0.5% EP during the first 2 weeks of life provided significant weight gain advantage compared to live vaccination alone. This advantage persisted through 2 weeks of EP withdrawal and subsequent challenge infection. EP supplementation also significantly lowered total lesion scores but did not significantly modify the effects of vaccination and challenge on plasma carotenoids or NO2-+NO3-. PMID- 12898223 TI - Shigella flexneri 2a strain 2457T expresses three members of the H-NS-like protein family: characterization of the Sfh protein. AB - Shigella flexneri 2a is known to express the H-NS nucleoid-structuring protein and the paralogous protein StpA. Using bioinformatic analysis we have now discovered a third member of the H-NS protein family, Sfh (Shigella flexneri H-NS like protein), in strain 2457T. This protein is encoded by the sfh gene, which is located on a high-molecular-mass plasmid that is closely related to the self transmissible plasmid R27. When expressed in Escherichia coli, the Sfh protein can complement an hns null mutation, restoring wild-type Bgl, porin protein, and mucoidy phenotypes, and wild-type expression of the fliC and proU genes. While a knockout mutation in the sfh gene alone had no effect on the expression of virulence genes in S. flexneri, an additive effect on virulence gene derepression was seen when the sfh lesion was combined with a mutation in hns. Over-expression of the sfh gene repressed expression of the VirB virulence regulatory protein and transcription of a VirB-dependent structural gene promoter. The purified Sfh protein bound specifically to DNA sequences containing the promoters of the virF and virB virulence regulatory genes. These findings show that Sfh has the ability to influence genetic events beyond the genetic element that encodes it, including the expression of the S. flexneri virulence genes. They raise the possibility of a triangular relationship among three closely related proteins with broad consequences for genetic events in the bacterium that harbours them. PMID- 12898225 TI - Cloning and characterization of Taenia saginata paramyosin cDNA. AB - A lambdaZAP-express cDNA library of Taenia saginata metacestodes was constructed. Antibody screening yielded a clone with an insert of 3,408 bp, an open reading frame of 2,589 bp, a deduced sequence of 863 amino acid and a molecular mass of 98.89 kDa. Alignments of the predicted amino acid sequence showed identity with paramyosins from several species: 98.8% with Taenia solium, 96.3% with Echinococcus.granulosus and about 70% with Schistosoma spp. The insert was expressed and purified. A collagen binding assay was performed which showed that T. saginata GST-paramyosin retained this property in a dose-dependent manner. Problems were encountered due to high backgrounds in serological assays in the homologous T. saginata system. However, the recombinant paramyosin was recognized by antibodies present in 31.6% of sera from T. solium seropositive cysticercosis patients and 100% of the sera from acute cysticercosis patients. The immunodominant epitope was the carboxyl-terminal fragment of the molecule. PMID- 12898226 TI - Human serum antibodies against shared antigens of different stages of Trichinella spiralis: relevance of glycan and protein epitopes. AB - Taking into account that antibodies against the surface antigens of newborn larvae (anti-NBL Abs) present in sera from individuals with chronic trichinellosis recognize antigenic determinants of the excretory-secretory muscle larva products (ML-ESP), and that these products are mainly glycoproteins, the aim of this work was to assess the frequency of anti-NBL Abs in sera from individuals with acute and chronic trichinellosis, to analyse the relevance of glycan and protein epitopes of the ML-ESP in the cross-reactivity phenomenon, and its correlation with the host's serum response towards these products. Anti-NBL surface Abs were determined in sera by indirect immunofluorescence. The degree of recognition by serum and purified anti-NBL Abs was evaluated comparatively before and after chemical deglycosylation of ML-ESP by immunoelectrotransfer blot assay. Results showed that 64% of the sera from individuals with acute trichinellosis and 35% of those belonging to the chronic phase had anti-NBL Abs, and also that the protein epitopes are the major ones responsible for the cross-reactivity phenomenon involving the ML-ESP and the NBL surface during both phases of the infection, while glycan epitopes are immunodominant in the stimulation of the host's immune system. A modulatory phenomenon in the immune response generated towards Trichinella spiralis NBL driven by the ML-ESP is postulated. PMID- 12898227 TI - Descriptions of two new species of the genus Tachygonetria Wedl, 1862 (Nematoda Pharyngodonidae) and discussion of the relationships among the species of the genus. AB - Two species of pinworm, parasites of Testudo graeca Linneaus, 1758 from Morocco, are described: Tachygonetria poulini sp. nov. and Tachygonetria settatensis sp. nov. Light and scanning electron microscopy studies revealed morphological differences in the structure of the cephalic end in both sexes, as well as in the structure of the caudal end in the male, which enabled us to differentiate these species from others, and from each other. The relationships among the species of the genus Tachygonetria are discussed. PMID- 12898228 TI - Ultrastructure of pigmented and unpigmented photoreceptors in cercariae of Trichobilharzia ocellata (Plathelminthes, Trematoda, Schistosomatidae): evidence for the evolution of parasitism in Neodermata. AB - The lens-covered pigment cup ocelli in free-swimming cercariae of Trichobilharzia ocellata were re-examined and the differentiation of the lenticular elements was studied in cercariae still enclosed in sporocysts. Each eye consists of a single rhabdomeric sensory cell, a single cup cell harboring pigment granules and the lens. This lens is developed step by step by the fusion of numerous dark platelets not enclosed by a bordering membrane. The material is presumably proteinaceous. There is no evidence that the lenses are of mitochondrial origin. Besides the pigment cup ocelli, a special type of unpigmented rhabdomeric photoreceptor was discovered. Three unicellular photoreceptors arranged in a three-dimensional configuration exist. These light-sensing organs show a principle of construction similar to that of phaosomous photoreceptors. We hypothesize that both types of photoreceptors are relevant for distinct reactions in the host-finding behaviour of the cercariae. The special type of lensing in the pigmented eyes seems to be restricted to Trichobilharzia species, whereas the phaosomous-like receptors may be more widespread in members of the Schistosomatidae. The obvious absence of any mitochondrial lensing in T. ocellata, and probably all other species of the Digenea, may be correlated with the evolution of endoparasitism in the Trematoda. PMID- 12898229 TI - Cytokine responses in mice infected with Clonorchis sinensis. AB - FVB and BALB/c mice show different morbidity, development of Clonorchis sinensis, and pathological changes following C. sinensis infection. FVB mice are susceptible and BALB/c mice are relatively more resistant to C. sinensis infection. To investigate the relationship between cytokine reaction and susceptibility to C. sinensis infection in FVB and BALB/c mice, we described both the patterns and kinetics of Th1 cytokines and Th2 cytokines in spleen cell culture. Interleukin (IL)-4 and IL-10 cytokine production in the culture supernatants of the concanavalin-A-stimulated spleen cells increased at 2-3 weeks post-infection in both strains. IL-5 production increased between 2 and 5 weeks post-infection in both strains, and reached a peak level at 2 weeks post infection in BALB/c mice and 4 weeks post-infection in FVB mice. In contrast, gamma interferon (IFN-gamma) production decreased between 2 and 4 weeks in both strains. IL-2 production increased slightly in BALB/c mice following infection, but was unchanged in FVB mice. IL-4 production over preinfection levels was significantly higher in FVB mice, whereas IFN-gamma, IL-2, and IL-10 production were significantly higher in BALB/c mice. The levels of serum immunoglobulin E (IgE) and blood eosinophils in both mouse strains significantly increased between 3 and 6 weeks postinfection. Serum IgE levels were significantly higher in FVB mice than in BALB/c mice. The results of this study suggest that susceptibility to C. sinensis infection is associated with Th2 cytokine production, especially IL-4 which is predominant in relatively susceptible FVB mice. PMID- 12898230 TI - Ultrastructure of the body wall of Cystidicoloides ephemeridarum (Nematoda, Cystidicolidae) in relation to the histopathology of this nematode in salmonids. AB - Light, scanning and transmission electron microscope examinations of adult Cystidicoloides ephemeridarum nematodes, parasitic in the stomach of brown trout (Salmo trutta fario L.), revealed that the body wall is composed of a cuticle, a hypodermis and longitudinally oriented somatic musculature. The body cuticle is composed of a trilaminated epicuticle, an outer and inner cortical zone, a median zone with globular bodies and a basal zone consisting of three subzones. Massive cuticle in the three zones and an osmiophilic lining is present in the buccal capsule. Armament is absent both from the buccal capsule and the head end. The surface of the body is transversely striated with cortical annuli. The only specialized attachment devices of C. ephemeridarum are the flexible overlapping margins of the annuli which are elongated in the first third of the body. Together with the sucking pressure of the oesophagus, they seem to play a role in the penetration and mechanical damage of the host's tissue. The hypodermis appears to be syncytial. Up to 300 worms were present in the bolus of food consumed by the host, or attached by their head ends in the lamina epithelialis, which resulted in localized disruptions of the mucosal epithelium with no inflammation. Infection with C. ephemeridarum seems to be a temporary, season dependent event of mild pathogenicity. PMID- 12898231 TI - Detection of P-glycoprotein-mediated multidrug resistance against anthelmintics in Haemonchus contortus using anti-human mdr1 monoclonal antibodies. AB - The "multidrug resistance" (MDR) system involves the action of transmembrane P glycoproteins (Pgp) which may be responsible for failure of chemotherapy in both invertebrates and vertebrates. We previously obtained partial reversion of anthelmintic resistance in nematodes subjected to both anthelmintics and inhibitors of this system. The results presented here are able to describe more accurately the presence of Pgp in nematodes because of the use of C219 and UIC2 monoclonal antibodies, which are used for the detection of human and mouse mdr1 gene products. These antibodies demonstrated the presence of Pgp in eggshells. Their role in these structures, which are considered to be passive barriers, remains to be determined. Flow cytometry analyses of the UIC2 staining allowed determination of the resistance of individuals, which varied within the parasite population. UIC2 demonstrated both the presence and activity of Pgp in nematodes as has previously been shown in tumour cells. Resistance seems to be due to an increase in both the number of Pgp sites and parasites with high levels of Pgp. PMID- 12898232 TI - Surgery for large primary liver cancer more than 10 cm in diameter. AB - PURPOSE: Large primary liver cancer (PLC) more than 10 cm in diameter is not infrequently encountered in clinical practice. This study evaluated the clinicopathological features and long-term results after surgery for large PLC. METHODS: Comparison of clinicopathological data between patients with PLC >/=10 cm ( n=1,227) and PLC <10 cm ( n=2,349) during the same period. RESULTS: In comparison with patients with PLC <10 cm, patients with PLC >/=10 cm were significantly younger ( P<0.01), had a lower incidence of asymptomatic tumors (9.1% vs 39.5%, P<0.001), higher alpha-fetoprotein levels ( >400 ng/ml, 78.3% vs 49.2%, P<0.001), higher gamma-glutamyl transpeptidase levels ( >6U, 87.7% vs 70.5%, P<0.001), a lower incidence of a history of hepatitis (45.0% vs 61.4%, P<0.001) and associated macronodular cirrhosis (cirrhotic nodules >/=0.3 cm, 59.8% vs 66.6%, P<0.001), poor differentiation of tumor cells (Edmondson grade 3 4, 24.3% vs 19.7%, P<0.01), a lower percentage of single nodule tumors (59.9% vs 75.4%, P<0.001) and well-encapsulated tumors (28.5% vs 62.1%, P<0.001), a higher proportion of tumor emboli in the portal vein (20.5% vs 9.0%, P<0.001), a lower resection rate (50.6% vs 86.8%, P<0.001), a lower curative resection rate (54.8% vs 78.3%, P<0.001), a higher operative mortality rate (4.5% vs 2.3%, P<0.001), and less local resection (52.5% vs 80.2%, P<0.001). The 5- and 10-year survival rates after resection were 26.2% and 17.5%, respectively, for patients with PLC >/=10 cm ( n=621), and 54.3% and 39.5%, respectively, for patients with PLC <10 cm ( n=2039) ( P<0.01). CONCLUSIONS: Large PLC had specific clinicopathological features. Surgery is the first choice of treatment. In selected patients, resection is safe and offers the chance of long-term survival. Large PLC does not exclude the possibility of cure. PMID- 12898234 TI - Naturally occurring lignans efficiently induce apoptosis in colorectal tumor cells. AB - Plant-derived lignans caused cell loss by apoptosis in colorectal adenoma and carcinoma cells. Nordihydroguaiaretic acid (NDGA), commonly used for the inhibition of lipoxygenase isoenzymes, showed the strongest growth inhibition with an IC50 of 1.9+/-0.5 microg followed by epiashantin (IC50=9.8+/-4.5 microM) and arctigenin (IC50=16.5+/-8.5 microM). The lignans caused a time- and dose dependent loss of mitochondrial membrane potential (MMP), down regulation of the anti-apoptotic protein bcl(xl) and an increase of the apoptotic index. The time interval until loss of MMP and down modulation of bcl(xl) became evident correlated with the efficiency of growth inhibition by NDGA, epiashantin and yangambin. Bcl2 and caspase 3 were not involved. NDGA also induced a shift of the culture population to the G2/M phase of the cell cycle. With respect to these results, naturally occurring lignans could be useful in the therapy and chemoprevention of colorectal tumors. PMID- 12898233 TI - Dendritic cell-based tumor vaccine for cervical cancer II: results of a clinical pilot study in 15 individual patients. AB - PURPOSE: Human papillomavirus (HPV) type 16 and 18 are the most prevalent genotypes in cervical cancer. The viral oncoproteins E6 and E7 are considered to be tumor-specific targets for immunotherapy. HPV E7 antigen-loaded autologous dendritic cells (DC) were evaluated as cellular tumor vaccine in a case series of cervical cancer patients. METHODS: Autologous monocyte-derived DCs were pulsed with recombinant HPV16 E7 or HPV18 E7 oncoprotein and administered to 15 stage IV cervical cancer patients. Safety, toxicity, and induction of serological and cellular immune responses were monitored. RESULTS: The vaccine was well-tolerated and no local or systemic side effects or toxicity were recorded. A specific serologic response was seen in 3/11 evaluated patients. Specific cellular immune responses (4/11) were detected with 2/10 positive de novo reactions plus one boosted preexistent response in proliferation assays and 3/11 in IFN-gamma ELISpot assays. A transient drop in tumor marker SCC was observed in 5/9 evaluable patients but did not correlate with markers of the immune response. No objective clinical response was observed. Tumor biopsies available from four patients showed severe or complete loss of HLA expression in three of the advanced tumors. CONCLUSION: Autologous dendritic cells pulsed with HPV E7 protein can induce T cell responses in a portion of late stage cervical cancer patients. Boosting of immune responses by adjuvants and vaccination of tumor HLA positive patients will be mandatory in future trials. PMID- 12898235 TI - Expression of caveolin-1 in gastrointestinal and extraintestinal cancers. AB - Caveolae are uniform vesicular invaginations of the cell membrane. Caveolin-1 is responsible for the formation of caveolae and plays a key role in membrane traffic and signal transduction. The contribution of caveolin-1 to carcinogenesis has been widely investigated; however, the expression pattern of caveolin-1 is controversial both in gastrointestinal and extraintestinal cancers. Most of the results based on cancer cell line experiments suggest that caveolin-1 might act as a tumor-suppressor gene. On the contrary, several studies on the expression of caveolin-1 in tumor tissues indicate a possible tumor-promoting effect of caveolin-1. In this article we summarize the divergent results of caveolin-1 expression in gastrointestinal and extraintestinal cancer regarding possible future therapeutic implications. PMID- 12898237 TI - Fructose 1,6 diphosphatase deficiency in a Turkish infant. PMID- 12898238 TI - Acquired angioedema and Helicobacter pylori infection in a child. AB - We describe a 10-year-old boy with acquired Helicobacter pylori infection and simultaneous angioedema which is a rare but life-threatening condition. Our patient was hospitalised with generalised angioedema and severe circulatory shock due to extreme loss of fluids and proteins into interstitial tissues (weight gain 10 kg within 2 days, extreme haemoconcentration--haemoglobin 206 g/l, haematocrit 0.570, leucocytosis 18,300 /microl, high lactate 13.8 mmol/l) and simultaneous failure of the complement system (C3 <0.16 g/l, C4 <0.13 g/l, CH50 45 U/ml, i.e. 50% of normal value, C1 inhibitor 0.21 g/l at the lower limit). All possible known causes of angioedema were excluded (infection, allergy, auto-immune disease, NSAIDs, lymphoproliferative disease) except for the simultaneous H. pylori infection which was proven serologically and histologically. Eradication therapy led to a complete remission of the H. pylori infection. An absence of angioedema and the restoration of the complement system was later observed. To the best of our knowledge, no similar case report of a child has yet been published. CONCLUSION: Helicobacter pylori infection should be considered in the development of angioedema in childhood. PMID- 12898236 TI - Quantitative reverse transcriptase polymerase chain reaction analysis for KiSS-1 and orphan G-protein-coupled receptor (hOT7T175) gene expression in hepatocellular carcinoma. AB - PURPOSE: KiSS-1 has been cloned as a human metastasis suppressor gene and an orphan G-protein-coupled receptor (hOT7T175) identified as the endogenous receptor of the KiSS-1 product. In the present study, we evaluated the clinical importance of KiSS-1 and hOT7T175 gene expression in hepatocellular carcinoma (HCC). METHODS: The expression levels of KiSS-1, hOT7T175 and glyceraldehyde-3 phosphate dehydrogenase (GAPDH) messenger RNAs (mRNAs) were analyzed quantitatively by real-time reverse transcriptase polymerase chain reaction (RT PCR) in 60 surgically resected HCCs. The KiSS-1/GAPDH and hOT7T175/GAPDH ratios of tumors were compared with clinicopathological findings. RESULTS: Loss of KiSS 1 mRNA expression was not detected in HCCs. The mean KiSS-1/GAPDH ratio did not change between non-cancerous cirrhotic livers and carcinomas. On the other hand, the average hOT7T175/GAPDH ratios increased from non-cancerous livers (0.08) to carcinomas (0.48). Overexpression of KiSS-1 and hOT7T175 genes was recognized in 6 tumors, which were in an advanced stage and showed poor survival. CONCLUSION: Overexpression of KiSS-1 and hOT7T175 genes was frequently observed and correlated with HCC progression; thus, the possibility that overexpressed KiSS-1 and hOT7T175 peptides mediate growth signals into cancer cells in HCCs is suggested. PMID- 12898240 TI - Surfactant therapy in neonates with respiratory failure due to haemorrhagic pulmonary oedema. AB - We studied the clinical and biochemical factors associated with surfactant dysfunction and factors affecting the responsiveness to exogenous surfactant among 27 neonates with haemorrhagic pulmonary oedema (HPE). HPE was defined as the presence of a large amount of blood-stained lung effluent and respiratory failure which was difficult to differentiate from respiratory distress syndrome. Among the neonates, 33% had very low birth weight, 96% were preterm, 70% were delivered by caesarean section, and 44% had delivery room intubation. The onset of HPE was at 1.5+/-0.1 h (mean +/- SEM) after birth. In 26 cases, surfactant was administered at 3.0+/-1.3 h after the onset of HPE. The concentrations of surfactant protein A (SP-A), disaturated phosphatidylcholine (DSPC), and albumin in the epithelial lining fluid were determined using the first lung effluent from the patients. The level of inhibitory activity against pulmonary surfactant in the effluent was determined in vitro. Surfactant inhibitory activity was associated with lower birth weight, earlier gestational age, delivery room intubation, earlier onset of HPE, and lower SP-A or DSPC concentration. A good response to exogenous surfactant, which was defined as ventilatory index <0.047 at 1 h after surfactant administration, was seen in 82% of cases, and was associated with lower serum albumin, lower birth weight, and earlier gestational age. Cases with higher DSPC concentration prior to surfactant administration and shorter interval between the onset of HPE and surfactant administration showed an immediate response to surfactant, followed by no increase in ventilatory index for 24 h after surfactant administration. CONCLUSION: exogenous surfactant appeared to be a useful adjunctive therapy for overcoming surfactant inhibition and normalising the respiratory status of infants with haemorraghic pulmonary oedema. Surfactant treatment for this indication awaits further investigations including a randomised controlled study. PMID- 12898241 TI - Primary Sjogren syndrome in the paediatric age: a multicentre survey. AB - Primary Sjogren syndrome (SS) is very rare in childhood. We collected a series of primary paediatric SS cases from different centres. A data collection form was prepared and sent to rheumatologists who were willing to participate. Data on 40 cases of primary SS with onset before the 16th birthday were collected. Almost all patients (35/40) were females, age at onset varied from 9.3 to 12.4 years (mean 10.7 years). Signs and symptoms at disease onset were mainly recurrent parotid swelling followed by sicca symptoms. Abnormal laboratory tests were found in the majority of cases. Regarding treatment, 22 patients were treated at some time with oral corticosteroids, seven with non-steroidal anti-inflammatory drugs, and five with hydroxychloroquine; two patients needed cyclosporine and one cyclophosphamide. Follow-up varied from 0 to 7.5 years from onset, without major complications in the majority of patients. CONCLUSION: recurrent parotid swelling is a common feature of primary Sjogren syndrome in childhood and often occurs as a presenting feature. Sicca symptoms may be rarer. PMID- 12898242 TI - Experience of continuous haemodiafiltration in a male neonate with ornithine transcarbamylase deficiency. PMID- 12898243 TI - Mechanisms of death in the early postoperative period following coronary artery bypass grafting for acquired heart disease. A clinicopathological study of 32 cases. AB - OBJECTIVES: A retrospective cardiopathological and clinical study was conducted in order to determine causes of perioperative death following coronary artery bypass grafting (CABG). EXPERIMENTAL DESIGN: Between January 1992 and June 1995, a total of 5749 CABG procedures were performed at the Heart Center Duisburg (Germany). Following the procedures, 218 patients died in hospital (mortality rate 3.8%). Fifty-eight were autopsied at the Institute of Pathology, Bethesda Hospital, Duisburg, and 32 autopsied cases were amenable to our study. Basis for selection was accessibility of clinical and morphological data and a postoperative death within 30 days. METHODS: In each case, morphological analysis of the heart and an evaluation of surgical and clinical data were performed in order to draw a conclusion on the mechanism of death. RESULTS: Using criteria defined by us, the following causes of death were determined: (1) surgical complications (43%); (2) severe coronary artery disease with incomplete revascularization (41%); (3) congestive heart failure (13%); (4) non-cardiac complications (3%). CONCLUSION: Criteria defined in this study may be useful in evaluations of causes of death after open heart surgery and may help to compare results in future series. Determination of the cause of death is important for the cardiac surgeon to reconsider indications and quality of surgical procedure. PMID- 12898244 TI - CD34+ fibrocytes in tubular carcinomas and radial scars of the breast. AB - The present study was undertaken to analyze whether and to what extent CD34+ fibrocytes and alpha-smooth muscle actin (alpha-SMA)-positive myofibroblasts occur in the stroma of radial scars and tubular carcinoma of the breast. We investigated a total of 24 radial scars obtained from 23 females and a total of 43 tubular carcinomas. All tubular carcinomas showed a virtually complete loss of CD34+ fibrocytes paralleled by a gain of alpha-SMA-positive myofibroblasts. The peripheral parts of radial scars harbored CD34+ fibrocytes comparable to normal breast tissue. In 23 of 24 radial scars, the central core was characterized by a loss of CD34+ fibrocytes accompanied by the presence of alpha-SMA-positive myofibroblasts in six cases, a pattern that up to now has only been described in malignant breast lesions. This finding underlines the close relationship between tubular carcinomas and radial scars. We conclude CD34- and alpha-SMA immunohistochemistry to be valuable adjunctive tools in distinguishing radial scars from tubular carcinomas. In the present study, the presence of CD34+ fibrocytes excluded malignancy. The absence of CD34+ fibrocytes paralleled by the presence of alpha-SMA myofibroblasts indicated malignancy in most cases, although it has to be carefully considered that in a minority of cases (6 of 24) the central core of radial scars may disclose this stromal composition. PMID- 12898245 TI - Periacinar retraction clefting in the prostatic needle core biopsies: an important diagnostic criterion or a simple artifact? AB - The diagnosis of prostatic adenocarcinoma in needle core biopsy is based on major and supportive criteria. One of the supportive criteria is the presence of retraction clefting around neoplastic glands. We analyzed a series of 137 prostatic cancer cases diagnosed by needle core biopsy to determine the frequency, extent and criteria for periacinar retraction clefting. Clefting was analyzed on ten neoplastic and ten normal glands in three different high power fields. One-third or more glands with clefts affecting more than 50% of circumference were significantly more common in tumors (51.8%) than in benign glands (8%) (P<0.0001). A stricter criterion that designated as positive the cases with at least 50% of neoplastic glands (15 of 30) with clefts that affected more than 50% of circumference revealed clefts in only 15.3% of the malignant cases but none in benign cases (0%) (P<0.0001). Regardless of their extension, 15 or more glands with clefts were also more prominent in malignant cases (86.9%) than in benign cases (20.4%) (P<0.0001). We conclude that periacinar retraction clefting represents a reliable criterion for diagnosis of the prostatic adenocarcinoma, especially in cases with clefts affecting more than 50% of circumference in at least 50% of suspicious glands. PMID- 12898246 TI - Histopathological parameters as predictors for the course of Crohn's disease. AB - BACKGROUND: Recent classification systems discriminate penetrating Crohn's disease (CD) from stricturing and non-stricturing/non-penetrating CD. As yet, no easily detectable marker is known that can predict the course of CD. Individual, clinical course-specific new treatment schemes would be highly desirable for different potentially divergent pathophysiological pathways (e.g., fistula vs stenosis). METHODS: Intestinal tissue biopsies from 63 CD patients with a disease follow-up of up to 7 years were studied retrospectively. In biopsy specimens, 34 histopathological features present prior to the onset of (a) strictures or (b) fistulas were evaluated and compared with biopsies from patients with non stricturing/non-penetrating disease. RESULTS: Five histomorphological parameters demonstrated significant associations to different disease courses when applying univariate analysis. In a multivariate logistic regression model (1) severe lymphocytic infiltration of the lamina propria, (2) presence of crypt atrophy, and (3) absence of lymphocytic infiltration of the epithelium are the best variables to predict an uncomplicated disease course (Nagelkerke R2=0.329; P=0.001). The combination of these parameters has a sensitivity of 67% and a specificity of 83% to predict non-stricturing/non-penetrating disease (positive predictive value=0.75). CONCLUSION: Histopathological parameters may help to predict complications of CD prior to their onset. The results of this study have to be confirmed prospectively. PMID- 12898247 TI - Loss of lamin A/C expression revealed by immuno-electron microscopy in dilated cardiomyopathy with atrioventricular block caused by LMNA gene defects. AB - Mutations of the LMNA gene encoding the lamin A and C nuclear envelope proteins cause an autosomal dominant form of dilated cardiomyopathy (DCM) with atrioventricular block (AVB). The aim of this study was to investigate ultrastructural nuclear membrane changes by conventional electron microscopy and protein expression by immuno-electron microscopy in the heart of patients with DCM and AVB due to LMNA gene mutations. Four immunohistochemical techniques were used: pre-embedding and post-embedding in Epon-Araldite resin and London Resin White (LRW), with and without silver enhancement. Parallel light microscopy immunohistochemistry studies were performed. Conventional electron microscopy showed a loss of integrity of the myocyte nuclei with blebs of the nuclear membrane, herniations and delamination of the nuclear lamina and nuclear pore clustering. Post-embedding LRW was the most informative technique for morphology and immuno-labelling. Immuno-labelling was almost absent in the nuclear envelope of patients with LMNA gene mutations, but intensely present in controls. The loss of labelling selectively affected myocyte nuclei; the endothelial cell nuclei were immunostained in patients and controls. Light immunohistochemistry confirmed the results. These findings confirm the hypothesis that LMNA gene defects are associated with a loss of protein expression in the selective compartment of non cycling myocyte nuclei. PMID- 12898248 TI - Disruption of the integrity of human peritoneal mesothelium by interleukin-1beta and tumor necrosis factor-alpha. AB - Inflammatory and neoplastic disease processes of the abdominal cavity are frequently associated with disruption of the integrity of the peritoneal mesothelium. In the present study, we analyzed the effects of the pro inflammatory cytokines interleukin-1beta (IL-1beta) and tumor necrosis factor alpha (TNF-alpha) on the morphology and expression of adhesion molecules of human peritoneal mesothelial cells (HPMC). Treatment of HPMC with IL-1beta and TNF alpha resulted in a time- and dose-dependent alteration of the normal cobblestone morphology of the mesothelium with loss of polarization, cellular retraction and exposure of the submesothelial matrix. The effect was already observable after 6 h of treatment and was most pronounced at a dose of 10 ng/ml of IL-1beta or TNF alpha. These morphological alterations were associated with a significant rearrangement of the expression of mesothelial adhesion molecules as detected by flow cytometry. IL-1beta and TNF-alpha both led to a loss of the expression of the hemidesmosomal integrin subunits alpha6 ( P<0.01 and P<0.001) and beta4 ( P<0.01) and an increased expression of the integrin subunit alpha5 ( P<0.001 and P<0.01). IL-1beta furthermore upregulated the expression of the integrin subunits alpha1, alpha2 and the adhesion molecule CD44 while the latter was downregulated by TNF-alpha. Our data indicate that IL-1beta and TNF-alpha may significantly affect disease processes of the abdominal cavity by their potential to disrupt the mesothelial basal cell-matrix adhesion and, thus, the integrity of the peritoneal mesothelial cell lining. PMID- 12898250 TI - The expression of gelatinase A (MMP-2) is required for normal development of zebrafish embryos. AB - Gelatinase A, also called matrix metalloproteinase 2 (MMP-2), belongs to the matrix metalloproteinase (MMP) family. MMP-2 cleaves type IV collagen, denatured collagen (gelatin), and other extracellular matrix (ECM) components. MMP-2 has been reported to be involved in a number of biological and pathological processes, but previous studies have not indicated that its expression is essential for early embryogenesis. In the current study, we have utilized zebrafish as a developmental model to study the role of MMP-2 during embryogenesis. We have successfully isolated a zebrafish MMP-2 (zMMP-2) homologue showing over 80% identity and over 90% similarity to its human counterpart. In situ analysis showed that zMMP-2 was expressed as early as the one-cell stage implying a maternal origin during oogenesis, and embryos continued to express zMMP-2 through at least the 72-h stage of development. RT-PCR analysis confirmed the in situ expression pattern and gelatin zymography indicated that a metalloproteinase with the same gel mobility as vertebrate MMP-2 was present in zebrafish embryos. Injection of zMMP-2 antisense morpholino oligonucleotides into 1- to 4-cell embryos resulted in a truncated axis, monitored through 72 h of development indicating that this metalloproteinase plays an important role in zebrafish embryogenesis. Monpholino-induced alterations in development began to be observed at 12 h of embryogenesis based on morphological and axis marker studies. The results obtained in zebrafish are in contrast to murine knockout studies that indicate that MMP-2 does not have a major role in mouse embryogenesis. PMID- 12898249 TI - Isolation and characterization of two T-box genes from sponges, the phylogenetically oldest metazoan taxon. AB - It is now well established that all metazoan phyla derived from one common ancestor, the hypothetical Urmetazoa. Due to the basal position of Porifera (Demospongiae) in the phylogenetic tree of Metazoa, studies on the mechanisms controlling the development of these animals can provide clues on the understanding of the origin of multicellular animals and on how the first organization of the body plan evolved. In this report we describe the isolation and genomic characterization of two T-box genes from the siliceous sponge Suberites domuncula. The phylogenetic analysis classifies one into the subfamily of Brachyury, Sd-Bra, and the second into the Tbx2 subfamily, Sd-Tbx2. Analyses of the Sd-Bra and Sd-Tbx2 sequences and their intron-exon structures demonstrate their basal position in the phylogeny of the T-box family, and allows us to hypothesize a model of the phylogenetic evolution of all T-box genes. Furthermore, we report the presence of two different products of alternative splicing of Sd-Bra, and demonstrate that they exist in different phosphorylation and glycosylation states in the sponge tissue. Sd-Bra expression in tissue and 3D cell aggregates (primmorphs) is analyzed, suggesting that Sd-Bra might also have a role in Porifera morphogenesis. PMID- 12898251 TI - Perceiving while acting: action affects perception. AB - In two experiments we studied how motor responses affect stimulus encoding when stimuli and responses are functionally unrelated and merely overlap in time. Such R-S effects across S-R assignments have been reported by Schubo, Aschersleben, and Prinz (2001), who found that stimulus encoding was affected by concurrent response execution in the sense of a contrast (i.e., emphasizing differences). The present study aimed at elucidating the mechanisms underlying this effect. Experiment 1 studied the time course of the R-S effect. Contrast was only obtained for short intertrial intervals (ITIs). With long ITIs contrast turned into assimilation (i.e., emphasizing similarities). Experiment 2 excluded an interpretation of the assimilation effect in terms of motor repetition. Our findings support the notion of a shared representational domain for perception and action control, and suggest that contrast between stimulus and response codes emerges when two S-R assignments compete with each other in perception. When perceptual competition is over, assimilation emerges in memory. PMID- 12898252 TI - DNA damage in the early primordial anther is closely correlated with stamen arrest in the female flower of cucumber (Cucumis sativus L.). AB - To investigate the regulatory mechanisms of sex expression in cucumber, morphological observations and biochemical analyses were carried out on inappropriate stamen development of female flowers of cucumber. It was found that developmental arrest of the inappropriate stamen mainly occurs at the anther primordium. This arrest is closely correlated with DNA damage, as detected by TUNEL assay, and might result from anther-specific DNase activation. It was also found that the DNA damage does not lead to cell degeneration, although chromatin condensation is observed in the anther primordia. PMID- 12898253 TI - Functional characterisation of two cytochrome b5-fusion desaturases from Anemone leveillei: the unexpected identification of a fatty acid Delta6-desaturase. AB - The Ranunculaceae are known to accumulate a wide range of unusual fatty acids in their seed lipids, and this variability has been advocated as a taxonomic marker. The Anemone species, Anemone leveillei L. and Anemone rivularis Buch.-Ham., have previously been reported to accumulate Delta5-desaturated fatty acids in their seed tissue [K. Aitzetmuller (1995) Plant Syst Evol 9:229-240]. Two cDNAs, AL1 and AL2, with similarity to plant cytochrome b5-fusion "front-end" desaturases were isolated from developing seeds of A. leveillei and their function identified by expression in Saccharomyces cerevisiae. AL2 was characterised as a sphingolipid long-chain-base Delta8-desaturase, while AL1 acted as a fatty acid desaturase. However, AL1 did not produce Delta5-desaturated fatty acids as expected; instead, when expressed in transgenic S. cerevisiae or Arabidopsis thaliana this enzyme was functionally characterised as a Delta6-desaturase. Northern analysis confirmed the expression of this gene in seed tissue and leaf tissue of A. leveillei, though Delta6-desaturated fatty acids were found to accumulate only in the leaf tissue. The unexpected characterisation of a Delta6 desaturase in A. leveillei has implications for the use of fatty acids in chemotaxonomic studies. This is also the first report of a higher-plant Delta6 desaturase from a family other than the Boraginaceae. PMID- 12898254 TI - Reduction in the endogenous arginine decarboxylase transcript levels in rice leads to depletion of the putrescine and spermidine pools with no concomitant changes in the expression of downstream genes in the polyamine biosynthetic pathway. AB - We investigated whether down-regulation of arginine decarboxylase (ADC) activity and concomitant changes in polyamine levels result in changes in the expression of downstream genes in the polyamine pathway. We generated transgenic rice (Oryza sativa L.) plants in which the rice adc gene was down-regulated by expression of its antisense oat (Avena sativa L.) ortholog. Plants expressed the oat mRNA adc transcript at different levels. The endogenous transcript was down-regulated in five out of eight plant lineages we studied in detail. Reduction in the steady state rice adc mRNA levels resulted in a concomitant decrease in ADC activity. The putrescine and spermidine pool was significantly reduced in plants with lower ADC activity. Expression of the rice ornithine decarboxylase (odc), S adenosylmethionine decarboxylase (samdc) and spermidine synthase (spd syn) transcripts was not affected. We demonstrate that even though levels of the key metabolites in the pathway were compromised, this did not influence steady-state transcription levels of the other genes involved in the pathway. Our results provide an insight into the different regulatory mechanisms that control gene expression in the polyamine biosynthetic pathway in plants by demonstrating that the endogenous pathway is uncoupled from manipulations that modulate polyamine levels by expression of orthologous transgenes. PMID- 12898255 TI - Plastids of three Cuscuta species differing in plastid coding capacity have a common parasite-specific RNA composition. AB - The chlorophyll containing holoparasitic species Cuscuta reflexa, the achlorophyllous species Cuscuta odorata and the intermediate species Cuscuta gronovii, which contains only traces of chlorophyll, were compared with respect to their plastid coding capacity and plastid gene expression at the level of RNA. While extensive deletions have taken place in the plastid DNA of the achlorophyllous species C. odorata, the green species C. reflexa has retained an almost complete plastid genome. Although the plastid genome of the intermediate species C. gronovii has suffered extensive deletions, in contrast to the plastid genome of C. odorata it has retained photosynthesis-related genes. Hybridization with radioactive 3'-labelled RNA revealed that in all three species only a small 'parasite-specific' portion of the plastid genome consisting of mainly rRNAs and tRNAs is represented at the level of steady-state RNA. Run-on transcription assays revealed that in plastids of C. reflexa the entire genome is transcribed. Hence, the subset of RNA species required for a parasitic lifestyle is preferentially stabilized in Cuscuta plastids. PMID- 12898256 TI - Loose-patch clamp currents from the hypothalamo-neurohypophysial system of the rat. AB - The loose-patch clamp technique was used to study voltage-activated currents from the surface of rat neurohypophysial and hypothalamic regions in situ. In the neurohypophysis, depolarizing pulses of 4-8 ms duration yielded tetrodotoxin (TTX)-sensitive sodium currents, a 4-AP-sensitive "A"-type potassium current, and a long-lasting outward TEA- and tetrandrine-sensitive Ca(2+)-activated potassium current. All of these currents were elicited during the application of the pulse. With high external calcium there were long-lasting inward currents blocked by Ni(2+) and Cd(2+), identifying them as voltage-gated calcium currents. Depolarizing pulses of 0.3-0.7 ms duration yielded fast biphasic responses, of 1 3 ms duration, composed of mostly sodium and "A"-type potassium currents. With high external calcium there were fast inward currents blocked by Ni(2+) and Cd(2+), indicating that these were voltage-gated calcium currents. These responses have the characteristics of action potential currents: they were elicited after the cessation of the applied pulse and the "A" component is eliminated together with the sodium component upon application of TTX. Similar responses to long and short pulses were obtained from the surface of the associated magnocellular somata in the supraoptic nucleus, and their projections. The explant currents are similar to those previously characterized using conventional methods from somata and terminals. PMID- 12898257 TI - Impaired slow inactivation due to a polymorphism and substitutions of Ser-906 in the II-III loop of the human Nav1.4 channel. AB - The loop connecting domains II and III of the sodium channel alpha-subunit is not known to have a major effect on channel gating. Recently mutations in the II-III loop of various sodium channel isoforms have been reported to cause channelopathies suggesting the functional importance of this region. In the II III loop of the skeletal muscle isoform Na(v)1.4, we found a Ser-to-Thr substitution at position 906 in 5% of patients with dyskalemic periodic paralysis but also in 4% of healthy human individuals. To investigate whether this position is important for channel gating, we characterized the following amino acids at 906 by whole-cell patch-clamp experiments: Gln, Ser, Thr, Cys, Pro, Val, ordered according to their hydrophobicity. All substitutions mainly affected slow inactivation. For example, Gln caused a +13-mV right-shift of the steady-state slow inactivation curve, and entry into slow inactivation was 6 times slower compared with Ser, leading to a destabilization of the slow inactivated state; in contrast, Val, at the other end of the hydrophobicity spectrum, shifted the steady-state slow inactivation curve by -6 mV and slowed the recovery from the slow inactivated state threefold compared with Ser, resulting in an enhancement of slow inactivation. Recovery from the slow inactivated state was also slowed by Pro, Cys and Thr. Our results suggest that (1) a benign polymorphism affects channel function, (2) the II-III loop is important for slow inactivation, and (3) the effects on slow inactivation may depend on the hydrophobicity of the residue at position 906. PMID- 12898258 TI - Recovery of cell volume and electrolytes of A6 cells after re-establishing isotonicity following hypotonic stress. AB - Cellular element concentrations and dry weight contents in A6 cells were determined using electron microprobe analysis to establish whether these cells exhibit a regulatory volume increase (post-RVD-RVI) when re-establishing isotonicity following a hypotonically induced regulatory volume decrease (RVD). Hypotonic stress was induced by reducing basolateral [NaCl], and hence, osmolarity fell from 260 to 140 mosmol/l. The alterations in cell volume after re establishing isotonicity, calculated from the cellular dry weight changes, indicate within the first 2 min cell shrinkage from 120 to 76% of control, compatible with almost ideal osmometric behaviour of A6 cells, and thereafter a post-RVD-RVI to 94%. The cellular uptake of osmolytes necessary to explain the post-RVD-RVI could be accounted for solely by a gain in cellular K and Cl. The involvement of a Na-K-2Cl cotransporter in most of the KCl uptake seems plausible since basolateral bumetanide blocked KCl uptake and post-RVD-RVI. The net uptake of cations (K uptake of 185.2, Na loss of 8.2 mmol/kg dry wt) during the isotonic period exceeded the Cl uptake by 38.2 mmol/kg dry wt, suggesting the uptake of another anion and/or the alteration of cellular buffer capacity. The relatively low Na concentration maintained during the isotonic period (13.3 vs. 20.4 mmol/kg wet wt under control conditions) might favour electrolyte uptake via the Na-K-2Cl cotransporter. PMID- 12898259 TI - Functional differences in type-I fibres from two slow skeletal muscles of rabbit. AB - The present study addressed the question of whether the slow fibres of mammalian skeletal muscle, containing the myosin heavy chain MHCI (type-I fibres), are a functionally homogeneous population. We compared various properties of Ca(2+) activated, skinned, type-I fibres from the soleus and semitendinosus muscles of a rabbit. Soleus type-I fibres showed significantly faster kinetics of stretch activation, measured as the time-to-peak of the stretch-induced, delayed force increase, t(3), than semitendinosus fibres (1239+/-438 ms, n=136, vs. 1600+/-409 ms, n=208 respectively) (means+/-SD, 22 degrees C). Similarly, the speed of unloaded shortening at 15 degrees C was faster in soleus than in semitendinosus fibres [0.79+/-0.16 fibre lengths (FL) s(-1), n=44, vs. 0.65+/-0.15 FL s(-1), n=35 respectively]. The kinetics of stretch activation were more temperature sensitive in semitendinosus than in soleus fibres. Finally, the generation of steady-state isometric force was more sensitive to Ca(2+) in semitendinosus than in soleus fibres: [pCa(50) (-log [Ca(2+)] for half-maximal activation) at 22 degrees C: 6.29+/-0.15, n=28, vs. 6.19+/-0.10, n=18 respectively]. These results suggest strongly that there is no functional homogeneity within type-I fibres of different muscles. The observed differences might reflect the existence of more than one functionally different slow myosin heavy chain isoforms or other modifications of contractile proteins. PMID- 12898260 TI - Retroviral endostatin gene transfer inhibits growth of human lung cancer in a murine orthotopic xenotransplant model. AB - BACKGROUND AND AIMS: The administration of endostatin, a potent anti-angiogenic agent, will be required for extended periods of time as a cancer treatment. The aim of the present study was to induce endogenous endostatin secretion in a continuous fashion, based on retroviral gene transfer. The tumor response was evaluated in an orthotopic murine tumor model of human lung cancer. MATERIALS AND METHODS: Human non-small-cell lung cancer cells (KNS 62) were retrovirally transduced with the human endostatin gene. An orthotopic murine xenotransplant model was used to investigate tumor growth, metastases and survival. After 4 weeks of subcutaneous growth, endostatin expression was measured by immunoblot analysis in tumor lysates. RESULTS: The growth of the subcutaneous tumors was significantly delayed, and orthotopic tumor growth and pleural metastases were significantly reduced in endostatin-transduced KNS 62 tumors. Prolongation of survival subsequent to orthotopic tumor induction was demonstrated. Strong endostatin expression was found in subcutaneous tumors after 4 weeks. CONCLUSION: Retroviral transduction of the human endostatin gene is capable of achieving long term endostatin expression. Endostatin transduction provides significant anti tumor effects with regard to local tumor growth, metastases and survival. PMID- 12898262 TI - Performing at extreme altitude: muscle cellular and subcellular adaptations. AB - This review reports on the collaborative efforts of the Department of Physiology of the University of Geneva headed by Paolo Cerretelli, the Research Institute at the Federal School of Physical Education in Magglingen and the Department of Anatomy of the University of Bern to elucidate the functional and structural conditions for and consequences of climbing successfully at altitudes in excess of 8000 m. Using a combination of physiological whole body measurements with biochemical, histochemical and morphometric analyses of muscle biopsy samples we were able to establish specific phenotypical alterations of muscle tissue exposed to extreme hypoxia and stress for prolonged periods of time. The decline in aerobic work capacity could be shown to be a consequence of a loss of muscle mass as well as of muscle tissue oxidative capacity whereby muscle capillarity was found to be maintained. The degradation of muscle tissue was further characterized by an increase in muscle lipofuscin. The latter is believed to be the consequence of lipid peroxidation eventually related to mitochondrial loss. Current work ensuing from our long-term collaboration suggests that Sherpas might be protected against the damaging effect of hypoxia by antioxidant mechanisms protecting their muscles under the conditions of extreme altitude. PMID- 12898263 TI - The optimal locomotion on gradients: walking, running or cycling? AB - On level ground, cycling is more economical than running, which in turn is more economical than walking in the high speed range. This paper investigates whether this ranking still holds when moving on a gradient, where the three modes are expected to be mainly facing the same burden, i.e. to counter gravity. By using data from the literature we have built a theoretical framework to predict the optimal mode as a function of the gradient. Cycling was found to be the mode of choice only below 10-15% gradient, while above it walking was the least expensive locomotion type. Seven amateur bikers were then asked to walk, run and ride on a treadmill at different gradients. The speed was set so as to maintain almost constant the metabolic demand across the different gradients. The results indicate that the "critical slope", i.e. the one above which walking is less expensive than cycling (and running), is about 13-15%. One subject was loaded during bipedal gaits with a bicycle-equivalent mass, to simulate to cross-country cycling situation. The critical slope was close to 20%, due to the higher metabolic cost of loaded walking and running. Part of the findings can be explained by the mechanically different paradigms of the three locomotion types. PMID- 12898264 TI - Impact of menstrual cycle phase on the exercise status of young, sedentary women. AB - The purpose of the present study was to compare exercise status during the follicular (FP) and luteal (LP) phases of the menstrual cycle of a single group of young, sedentary women, where the marked differential in the blood concentrations of 17beta-oestradiol ([E(2)]) and progesterone ([P(4)]) has the potential to alter the metabolic response to exercise. Fourteen females [21.8 (4.0) years, peak oxygen uptake ( VO(2peak)) <45 ml x kg (-1) x min(-1)] performed both incremental exercise to exhaustion and steady-state submaximal cycle ergometer exercise while measurements were made of several metabolic and hormonal variables. With the incremental exercise test, time to exhaustion, maximal power output and total work done were not different between the two phases, nor were the absolute values for VO(2peak) or the corresponding values for ventilation ( VE), respiratory frequency ( f(R)) and heart rate (HR). Resting, end-exercise and peak (post-exercise) plasma lactate concentrations and the lactate threshold were not different between the two phases either. However, as the workloads increased during the incremental protocol, plasma lactate concentration, carbon dioxide output ( VCO(2)) and the respiratory exchange ratio (RER) all were lower during LP, while oxygen uptake ( VO(2)) was higher. With steady-state submaximal exercise, at workloads corresponding to 25% and 75% of menstrual cycle phase-specific VO(2peak), VO(2) and the oxygen pulse ( VO(2)/HR) were higher and RER and plasma lactate concentration lower during LP. Regardless of phase, [E(2)] increased with both incremental and steady-state submaximal exercise, while [P(4)] was unchanged. It is concluded that while exercise capacity, as defined by VO(2peak) and the lactate threshold, is unaffected by cycle phase in young, sedentary women, the metabolic responses in the LP during both incremental and steady-state submaximal exercise suggest a greater dependence on fat as an energy source. PMID- 12898265 TI - The metabolic transition speed between backward walking and running. AB - Although the metabolic transition speed for forward exercise has already been determined, the walk-run transition speed for backward exercise has not been investigated before. The aim of this study was to determine the speed at which it becomes metabolically more efficient to run backwards than to walk backwards. Eighteen healthy volunteers, who successfully completed three backward exercise practice sessions, participated in the study. All subjects randomly performed two exercise tests: backward walking and backward running. Both protocols started at a treadmill speed of 5 km(.)h(-1). Every minute the speed was increased by 0.5 km(.)h(-1) until 8 km(.)h(-1) was reached. Cardiorespiratory variables were continuously measured and blood lactate concentration [La] was determined every 2 min, using the Accusport lactate analyser. At each work load subjects rated their perceived exertion (RPE), using the Borg scale. There were no statistically significant differences in oxygen consumption, minute ventilation and heart rate between 6 and 7 km(.)h(-1), for backward walking and backward running ( P>0.05). There was no statistically significant difference in blood [La] between walking and running at 7.5 km(.)h(-1) ( P>0.05). According to the RPE values, subjects rated running at speeds less than 6 km(.)h(-1) more difficult than walking at similar speeds. We conclude that the metabolic transition speed between backward walking and running is between 6 and 7 km(.)h(-1), which is lower than the metabolic transition speed for forward locomotion (7.2-7.9 km(.)h(-1)). PMID- 12898266 TI - Growth hormone substitution titrated to obtain IGF-I levels in the physiological range in hypopituitary adults: effects upon dynamic strength, endurance and EMG. AB - We studied the effects of individualised growth hormone (GH) substitution, aiming at normal insulin-like growth factor I (IGF-I) levels, on biomechanical output and surface electromyogram (EMG) of isokinetic muscle strength and endurance performance in 18 hypopituitary adults and compared with 17 matched healthy controls. The muscle function tests consisted of isokinetic contractions of the right knee extensors, from which torque and EMG were recorded. Three patients were excluded from the final analysis of the muscle function tests due to technical errors and one control subject moved from the area during the study. We found that GH-deficient adults without GH substitution were weaker and had less endurance than healthy control subjects. At the group level, plasma levels of IGF I were normalised but generally no significant effects upon biomechanical output and EMG were found after dose titration and 6 months of a constant GH dose. However, subjects with the largest changes in IGF-I had significantly better biomechanical output and EMG compared to those with small changes in IGF-I. This finding may indicate that the net increase in IGF-I levels is critical for improvements in biomechanical output, EMG and perception of fatigue to occur. PMID- 12898267 TI - Decrease in peak heart rate with acute hypoxia in relation to sea level VO(2max). AB - The aim of this study was to evaluate the influence of arterial oxygen saturation ( SaO(2)) on maximal heart rate during maximal exercise under conditions of acute hypoxia compared with normoxia. Forty-six males were divided into three groups depending on their sea level maximal oxygen consumption ( VO(2max)): high [GH, VO(2max)=64.2 (3.3) ml x min(-1) x kg(-1)], medium [GM, 50.8 (3.9) ml x min(-1) x kg(-1)] and low [GL, 41.0 (1.9) ml x min(-1) x kg(-1)]. All subjects performed a maximal exercise test in two conditions of inspired oxygen tension ( PIO(2), (149 mmHg and 70 mmHg). Among the GM group, seven subjects performed five supplementary incremental exercise tests at PIO(2) 136, 118, 104, 92, and 80 mmHg. Measurements of VO(2max) and SaO(2) using an ear-oxymeter were carried out at all levels of PIO(2). The decrease in SaO(2 )and peak heart rate (HR(peak)) with PIO(2) became significant from 104 and 92 mmHg. SaO(2) correlated with the decrease in HR(peak). For PIO(2)=70 mmHg, the decrease in VO(2max), SaO(2) and HR(peak) was, respectively, 44%, 62%, and 17.0 bpm for GH, 38%, 68%, and 14.7 bpm for GM, and 34%, 68%, and 11.8 bpm for GL. During maximal exercise in hypoxia, SaO(2) was lower for GH than GM and GL ( p<0.01). Among subjects in GH, five presented exercise-induced hypoxemia (EIH) when exercising in normoxia. The EIH group exhibited a greater decrement in HR(peak) than the non-EIH group at maximal hypoxic exercise (21.2 bpm vs. 15.0 bpm; p<0.05). When subjects are exposed to acute hypoxia, the lower SaO(2), due either to lower PIO(2) or to training status, is associated with lower HR(peak). PMID- 12898268 TI - Effects of the order of running and cycling of similar intensity and duration on pulmonary diffusing capacity in triathletes. AB - To study the pathophysiological mechanisms involved in the decrease of post triathlon diffusing capacity (DLco), blood rheologic properties (blood viscosity: eta(b); changes in plasma volume: deltaPV) and atrial natriuretic factor (ANF) were assessed in ten triathletes during cycle-run (CR) and run-cycle (RC) trials at a metabolic intensity of 75% of maximal oxygen consumption ( VO(2max)). The DLco was measured before and 10 min after trials. ANF and deltaPV were measured at rest, after the cycle and run of CR and RC trials, and at the end of and 10 min after exercise. RC led to a greater deltaDLco decrease, a lower ANF concentration and a lower deltaPV than did CR, whereas for both CR and RC eta(b) was increased throughout exercise and 10 min after. In addition, after CR the deltaDLco decrease was inversely correlated ( r=-0.764; P<0.01) with deltaPV. The association of decreased plasma volume, increased eta(b), and lower ANF concentrations after RC suggested that lower blood pulmonary volume may have caused the greater decrease in Dlco as compared with CR. The inverse correlation between deltaPV and deltaDLco reinforces the hypothesis that fluid shifts limit the post-exercise DLco decrease after the CR succession in triathletes. Lastly, cycling in the crouched position might increase intra-thoracic pressure, decrease thorax volume due to the forearm position on the handlebars, and weaken peripheral muscular pump efficacy, all of which would limit venous return to the heart, and thus result in low pulmonary blood volume. Compared with cycling, running appeared to induce the opposite effects. PMID- 12898269 TI - Effect of side dominance on myoelectric manifestations of muscle fatigue in the human upper trapezius muscle. AB - The purpose of this study was to investigate whether differences in the peripheral and control properties of the neuromuscular system due to long-term preferential use, related to side dominance, affect postural muscles, such as the upper trapezius. Therefore, fatigability properties of the upper trapezius muscles of the dominant and non-dominant side were assessed. Surface EMG signals were detected from the upper trapezius muscles of both sides of nine right- and five left-handed subjects with adhesive linear electrode arrays consisting of eight contact points. Static constant force contractions with the arms 90 degrees abducted were performed by the subjects while holding hand loads of 0 kg, 0.5 kg, and 1 kg. Surface EMG spectral and amplitude variables were computed from the recorded signals. EMG spectral variable rate of change (indicating fatigue) showed a statistically significant difference between the two sides, with the dominant side less fatigable than the non-dominant one. The observed differences held for both the right- and left-handed subject group. A possible explanation for the results is that long preferential use of one side with respect to the other leads to changes in muscle fiber membrane and control properties, in agreement with previous results on limb muscles. PMID- 12898270 TI - Critical review of the epidemiological literature on occupational exposure to perchloroethylene and cancer. AB - OBJECTIVES: Of an estimated 500,000 workers in the USA potentially exposed to perchloroethylene (PCE), the largest share is employed in the dry-cleaning industry. PCE, a non-flammable solvent, has commercial applications as a chemical intermediate, metal degreaser and, since the 1950s, primary solvent in the dry cleaning industry. The International Agency for Research on Cancer (IARC) currently finds sufficient evidence to designate PCE as carcinogenic in animals, with limited evidence in humans. With regard to occupational exposure through dry cleaning, PCE is considered to be possibly carcinogenic to humans. This review was conducted to assess the current epidemiological literature on PCE and specific cancers. METHODS: A comprehensive search was conducted to identify all available epidemiological literature pertaining to the carcinogenic effects of PCE. Forty-four papers that provided reasonable data on up to 17 cancer sites were critically reviewed in the context of the available background literature for each cancer site and were assessed on the basis of specified methodological and scientific quality criteria. RESULTS: While all the epidemiological studies selected for review investigated similar exposure-health outcome relationships, there was a broad diversity of proxy measurements of exposure to PCE, as well as numerous specific cancer outcomes of interest. The widespread lack of valid exposure measurements or other adequate indicators of potential for exposure were consistent limitations. We found no evidence of an association between breast, prostate, skin or brain cancer and exposure to PCE. A relationship between PCE and cancer of the following sites was considered unlikely: oral cavity, liver, pancreas, cervix lung. Scientific evidence was inadequate for laryngeal, kidney, esophageal and bladder cancers. CONCLUSIONS: The current epidemiological evidence does not support a conclusion that occupational exposure to PCE is a risk factor for cancer of any specific site. Priority areas in which additional data are most needed include cancers of the esophagus and bladder. PMID- 12898271 TI - Perceived muscular tension, emotional stress, psychological demands and physical load during VDU work. AB - OBJECTIVES: The aim of this study was to investigate whether perceived muscular tension, psychological demands and emotional stress were associated with physical load or working technique during visual display unit (VDU) work. METHODS: Subjects (28 women and 29 men) from two different organisations volunteered to participate in the study. The study design was cross-sectional, and the data were assessed when the subjects performed their usual work tasks at their usual work place. Multivariate linear and logistic regression models were used to investigate possible associations. The different outcome variables were: median muscle activity and muscular rest in the trapezius muscle bilaterally, wrist movements and working technique. The different explanatory variables were: perceived muscular tension (binary), emotional stress (binary), psychological demands (binary), organisation (binary) and gender (binary). Age (continuous) and present musculoskeletal pain (binary) were also controlled for in the multivariate models. Electromyography (EMG) and electrogoniometers were used to assess the physical load, and the data collection time was 15 min. An ergonomic checklist was used to assess working technique, i.e. work with lifted shoulders. RESULTS: Subjects who perceived muscular tension at least a few times per week the month before the measurements were made worked with higher muscle activity [expressed as per cent of a reference voluntary electrical activity (% RVE)] in the trapezius muscle bilaterally (5% RVE, P=0.05). High emotional stress during the measurement was associated with higher muscle activity in the trapezius muscle on the side not operating the computer mouse (8% RVE, P=0.006). Subjects who reported high levels of emotional stress worked more often with lifted shoulders (odds ratio 6.0, 95% CI 1.2-28.9). However, when present musculoskeletal pain was included in the multivariate model the odds ratio for high emotional stress decreased to 4.5 (95% CI 0.9-23.2). CONCLUSIONS: Perceived muscular tension and emotional stress were associated with physical load, in terms of muscle activity in the trapezius muscles, during VDU work in ordinary occupational settings. PMID- 12898272 TI - Nicotinic receptor alpha 7-subunits are coupled to the stimulation of nitric oxide synthase in rat dorsal root ganglion neurons. AB - In dorsal root ganglia (DRG) intraganglionic communication takes place both among neurons and between neurons and satellite cells. One diffusible substance involved in this signalling is nitric oxide (NO), and acetylcholine (ACh) is a candidate for the stimulation of intraganglionic NO synthesis. DRG neurons react to ACh-receptor stimulation with NO-dependent cGMP production. Here, we investigated the role of the alpha 7-subunit containing Ca(2+)-permeable nicotinic ACh receptors (nAChR) in this process. The alpha 7-nAChR mRNA and the protein were expressed in virtually all lumbar DRG neurons as evidenced by laser assisted cell picking and oligo cell RT-PCR, in situ hybridisation and immunohistochemistry. Strong alpha 7-nAChR immunoreactivity was present in vanilloid receptor 1-immunoreactive, i.e. nociceptive, neurons. A neuronal production of NO in response to nicotine could be demonstrated in DRG slice preparations utilising the NO-sensitive fluorescent indicator diaminofluorescein diacetate (DAF-2DA). This stimulation of NO production was sensitive to inhibition of alpha 7-nAChR by mecamylamine and alpha-bungarotoxin, to inhibition of nitric oxide synthase (NOS) with L-NAME and L-NMMA, and to the blockade of voltage-operated Ca(2+) channels by verapamil. The results show the presence of the alpha 7-nAChR subunit in nociceptive rat DRG neurons and provide evidence for its coupling to NOS activation, indicating a role of this pathway in the intraganglionic communication in sensory ganglia. PMID- 12898273 TI - Spatiotemporal expression patterns of sialoglycoconjugates during nephron morphogenesis and their regional and cell type-specific distribution in adult rat kidney. AB - The expression of alpha2,6- and alpha2,3-linked sialic acids on N-glycans was studied in embryonic, postnatal, and adult rat kidney. Histochemistry and blotting using Polyporus squamosus and Sambucus nigra lectins for alpha2,6-linked sialic acids and the Maackia amurensis lectin for alpha2,3-linked sialic acids were performed and sialyltransferase activity was assayed. N-glycans with alpha2,6- and alpha2,3-linked sialic acid were differently expressed in the two embryonic anlagen and early stages of nephron. Metanephrogenic mesenchyme was positive for alpha2,3-linked sialic acid but not for the alpha2,6-linked one, which became detectable initially in the proximal part of S-shaped bodies. Collecting ducts were positive for alpha2,6-linked sialic acid, whereas alpha2,3 linked sialic acid was restricted to their ampullae. Although positive in embryonic kidney, S1 and S2 of proximal tubules became unreactive for alpha2,3 linked sialic acid in postnatal and adult kidneys. In adult kidney, intercalated but not principal cells of collecting ducts were reactive for alpha2,3-linked sialic acid. In contrast, alpha2,6-linked sialic acids were detected in all cells of adult kidney nephron. Blot analysis revealed a different but steady pattern of bands reactive for alpha2,6- and alpha2,3-linked sialic acid in embryonic, postnatal, and adult kidney. Activity of alpha2,6 and alpha2,3 sialyltransferases was highest in embryonic kidney and decreased over postnatal to adult kidney with the activity of alpha2,6 sialyltransferase always being three to fourfold that of alpha2,3 sialyltransferase. Thus, alpha2,6- and alpha2,3-linked sialic acids are differently expressed in embryonic anlagen and mesenchyme-derived early stages of nephron and show regional and cell type-specific differences in adult kidney. PMID- 12898274 TI - Detection of HER2/neu gene amplification in archival paraffin-embedded breast cancer tissues by fluorescence in situ hybridization. AB - We evaluated HER2/neu gene amplification by fluorescence in situ hybridization (FISH) in archival paraffin-embedded breast cancer tissues. Tumors from 63 human invasive breast cancers were categorized into two groups depending on whether the paraffin-embedded tissue blocks had been stored for more or less than 12 months duration. These were subjected to routine and modified FISH protocols. As microwave oven formalin fixation of tissues was carried out in the majority of the older archived specimens, the effect of this fixation method was also analyzed. FISH signals were obtained in all 13 archival specimens stored for less than 12 months. However, in 50 specimens stored for more than 12 months duration, the procedure was successful in only 10 specimens (20%), for which the pretreatment procedure had to be individually optimized for each specimen. There was no significant difference in the detection of FISH signals between microwave oven and routinely fixed specimens. PMID- 12898275 TI - Expression of glutamate transporters in human and rat retina and rat optic nerve. AB - l-Glutamate is the major excitatory transmitter in the vertebrate retina and plays a central role in the transmission of the various retinal neurons. Glutamate is removed from the extracellular space by at least five different glutamate transporters. The cellular distribution of these has been studied so far mainly using immunocytochemistry. In the present study non-radioactive in situ hybridisation using complementary RNA probes was applied in order to identify the cell types of rat retina and optic nerve expressing generic GLT1, GLT1 variant (GLT1v or GLT1B), GLAST and EAAC1. The results were compared with immunocytochemical data achieved using affinity-purified antibodies against transporter peptides. In the immunohistochemical studies the human retina was included. The study showed that in the rat retina GLT1v and EAAC1 were coexpressed in various cell types, i.e. photoreceptor, bipolar, horizontal, amacrine, ganglion and Muller cells, whereas GLAST was only detected in Muller cells and astrocytes. In the rat optic nerve GLT1v and EAAC1 were preferentially expressed in oligodendrocytes, whereas GLAST was revealed to be present mainly in astrocytes. Generic GLT1 could not be detected in the retina or optic nerve. The cellular distribution of glutamate transporters (only immunocytochemistry) in the human retina was very similar to that of the rat retina. Remarkable results of our studies were that generic GLT1 was not detectable in the rat (and human) retina and that GLT1v and EAAC1 were demonstrable in most cell types of the retina (including photoreceptor cells and their terminals). PMID- 12898276 TI - Robert Feulgen Prize Lecture. Grenzganger: adult bone marrow cells populate the brain. AB - While the brain has traditionally been considered a rather secluded site, recent studies suggest that adult bone marrow (BM)-derived stem cells can generate glia and neurons in rodents and humans. Macrophages and microglia are the first to appear in the murine brain after transplantation of genetically marked BM cells. Within weeks after transplantation, some authors have found astrocytes and cells expressing neuronal antigens. We detected cerebellar Purkinje neurons and interneurons, such as basket cells, expressing the green fluorescent protein (GFP) 10-15 months after transplantation of GFP-labeled BM cells. The results push the boundaries of our classic view of lineage restriction. PMID- 12898277 TI - Robert Feulgen Lecture. Microscopic assessment of membrane protein structure and function. AB - Membrane proteins represent an important class of proteins that are encoded by about 40% of all genes, but compared to soluble proteins structural information is sparse. Most of the atomic coordinates currently available are from bacterial membrane proteins and have been obtained by X-ray crystallography. Recent results demonstrate the imaging power of the atomic force microscope and the accuracy of electron crystallography. These methods allow membrane proteins to be studied while embedded in the bilayer, and thus in a functional state. The low signal-to noise ratio of cryoelectron microscopy is overcome by crystallizing membrane proteins in a two-dimensional protein-lipid membrane, allowing its atomic structure to be determined. In contrast, the high signal-to-noise ratio of atomic force microscopy allows individual protein surfaces to be imaged at subnanometer resolution, and their conformational states to be sampled. This review discusses examples of microscopic membrane protein structure determination and illuminates recent progress. PMID- 12898278 TI - The effects of different intraocular irrigating solutions on the apoptosis of cultured RPE cells. AB - PURPOSE: To investigate the effect on porcine retinal pigment epithelial (RPE) cells of modified intraocular irrigating solutions compared with BSS and BSS plus. METHODS: Confluent cultures of RPE cells were incubated in experimental intraocular irrigating solutions. The cells were then examined for breakdown of DNA by the TUNEL procedure. Fragmentation of the DNA from cells was also confirmed by flow cytometry. RESULTS: Most of the experimentally, treated cells exhibited a shrunken appearance for up to 72 h. There was a steady increase in the number of cells labeled by the TUNEL method in three solutions with time. The data demonstrated that the influence of solutions on the percentage of RPE cell nuclei that gave a clear positive TUNEL stain was, in ascending order: modified solution>BSS>BSS plus. Of the three solutions tested, BSS plus showed least apoptosis. CONCLUSIONS: It is suggested that BSS plus is less harmful to RPE cells than the other solutions tested. Introduction of other adjunctive solutions of antibiotics, mydriatics, miotics, and steroid, resulting in an altered electrolyte balance, pH, or osmolality in the solution, may compromise the safety and efficacy of a properly formulated and packaged solution. PMID- 12898279 TI - Influence of argon laser trabeculoplasty on transforming growth factor-beta 2 concentration and bleb scarring following trabeculectomy. AB - BACKGROUND: The purpose of this study was to evaluate the influence of previous argon laser trabeculoplasty (ALT) on transforming growth factor-beta 2 (TGF-beta 2) concentration of the aqueous humor and its influence on bleb scarring after trabeculectomy. METHODS: Fifty-one patients with primary open-angle glaucoma (POAG) and 29 patients with exfoliation (XFS) glaucoma were recruited for this prospective study before undergoing trabeculectomy. Sixty to 200 micro l of aqueous humor were analyzed for total and biologically active TGF-beta 2 concentrations (R and D Systems). TGF-beta 2 levels and a standardized bleb assessment were compared between the ALT- and non-ALT-treated groups. RESULTS: POAG eyes without ALT showed significantly higher total TGF-beta 2 levels (2,317.7+/-1,041.1 pg/ml) than eyes with previous ALT (1,621.6+/-899.6 pg/ml; P=0.026). No significant difference was found for active TGF-beta 2 levels (ALT: 238.1+/-119.0 pg/ml; no ALT: 220.1+/-96.9 pg/ml; P=0.585). In XFS patients ALT did not alter total TGF-beta 2 levels (ALT: 1,524.9+/-624.9 pg/ml, no ALT: 1,220+/-499.1 pg/ml; P=0.20), but active TGF-beta 2 was significantly higher in the ALT-treated (237.0+/-99.7 pg/ml) than in the non-ALT-treated (140.0+/-95.3 pg/ml, P=0.028) group. Bleb grading revealed no statistical difference between the ALT- and non-ALT-treated groups in POAG (P=0.545, Fisher's exact test), whereas XFS patients with ALT were at increased risk for scarring compared to non ALT-treated patients (P=0.053). CONCLUSIONS: ALT appears to increase the risk of scarring in XFS patients because of increased levels of activated TGF-beta 2. PMID- 12898280 TI - Metipranolol attenuates lipid peroxidation in rat brain: a comparative study with other antiglaucoma drugs. AB - BACKGROUND: Free radical production seems to be involved in the pathogenesis of a number of ocular diseases. Certain beta-adrenoceptor antagonists display antioxidant properties, but these have not been ascribed to any of the presently used ophthalmic beta-adrenoceptor antagonists. Therefore, we examined the influence of ophthalmic beta-adrenoceptor antagonists and other antiglaucoma drugs on stimulated lipid peroxidation in rat brain homogenates. METHODS: Lipid peroxidation in rat brain homogenates was stimulated by iron/ascorbate or sodium nitroprusside. Lipid peroxidation was assessed by the formation of thiobarbituric acid reactive species (TBARS). RESULTS: Of the antiglaucoma drugs tested (brimonidine, carteolol, dorzolamide, latanoprost, levobetaxolol, levobunolol, metipranolol, pilocarpine, timolol, travoprost and unoprostone), only metipranolol and its active metabolite, desacetylmetipranolol, were found to significantly reduce iron/ascorbate-induced lipid peroxidation in rat brain homogenates with IC50 values of 6.9 and 1.1 microM, respectively. Metipranolol and desacetylmetipranolol also concentration-dependently inhibited sodium nitroprusside-stimulated lipid peroxidation in rat brain homogenates, displaying IC50 values of 25.1 and 2.6 microM, respectively. CONCLUSION: These data indicate that metipranolol and desacetylmetipranolol exhibit remarkable antioxidant properties, with an effect not dissimilar from the reference antioxidant trolox. PMID- 12898281 TI - Comparison between central corneal thickness and IOP in patients with macrodiscs with physiologic macrocup and normal-sized vital discs. AB - BACKGROUND: Macrodiscs with physiologic macrocups and elevated intraocular pressure (IOP) are frequently suspicious for glaucoma. Patients with an elevated c/d ratio and an IOP of more than 21 mmHg are often treated for glaucoma. We investigated a possible relationship between macrodiscs, IOP and central corneal thickness. METHODS: Using the Erlangen glaucoma registry, 341 out of 1,096 consecutive patients with unremarkable and repeated visual field (Octopus 500, G1 3, MD<2.1) were selected. Most of these patients had been sent as glaucoma suspects. The following data were collected: corneal ultrasound pachymetry (Tomey, AL-2000), IOP profile, visual acuity and central corneal power. The morphometric analysis was performed by planimetry using the Littmann formula. Optic nerve heads with a disc area of more than 3.1 mm(2) were defined as macrodiscs. RESULTS: Patients were subdivided into the following groups: group A: healthy macrodiscs, IOP <22 mmHg (87 patients); group B: healthy macrodiscs, IOP >21 mmHg (66 patients); group C: normal-sized healthy discs, IOP <22 mmHg (93 patients); group D: normal-sized healthy discs, IOP >21 mmHg (95 patients). Mean corneal thickness was: group A: 575+/-36 microm, group B: 596+/-37 microm, group C: 557+/-31 microm and group D: 585+/-38 microm ( P=0.003). Mean central corneal power measured: group A: 41.7+/-1.3 D, group B: 42.4+/-1.3 D, group C: 43.0+/-1.2 D and group D: 42.8+/-1.2 D ( P<0.05). The maximum of IOP was: group A: 17.6+/ 3.1, group B: 26.2+/-4.1, group C: 17.2+/-3.4 and group D: 29.7+/-5.5 mmHg. Optic disc size measurements were: group A: 3.91+/-0.66, group B: 3.65+/-0.6, group C: 2.56+/-0.30 mm(2) and group D: 2.39+/-0.39 microm, respectively. CONCLUSIONS: Macrodiscs with elevated IOP have a higher corneal curvature (mean: +0.7 D) and a thicker central cornea (mean: +21 microm) compared to macrodiscs without elevated IOP. An exact evaluation of optic disc morphology in combination with pachymetric and keratometric measurements in relation to IOP could avoid an overtreatment in patients with suspect primary open-angle glaucoma. PMID- 12898282 TI - Towards the cortical representation of form and motion stimuli generated by a retina implant. AB - BACKGROUND: A retina implant for restoring basic visual perception in patients who are blind due to photoreceptor loss should not only evoke focal phosphenes at high resolution, but should also generate cortical representations of form and motion. We are currently exploring these potential capabilities in anaesthetised cats. METHODS: Fibre electrodes were inserted through a small scleral incision onto the retinal surface for stimulation. For the recording of cortical population activities we placed up to 16 fibre electrodes in areas 17 and/or 18. Retinal and cortical electrodes were adjusted to corresponding sites, i.e., overlapping receptive fields. Electrical stimuli were charge-balanced impulses (200 micros, 10-100 microA). Basic form stimuli were generated by the selective and synchronous activation of some of the seven retinal stimulation electrodes. Movement stimuli were generated by spatio-temporal shifting of form stimuli. From multiple microelectrode recordings we computed stimulus-related spatio-temporal cortical activation profiles. We used these profiles to estimate the relations between stimulation distance and spatial resolution (form) and between stimulus velocity and spatio-temporal resolution (movement). Influences by the retino cortical pathway were assessed by comparing cortical activations evoked by true form or motion stimuli with synthetic responses composed by superpositioning of responses to appropriate subsets of form and motion stimuli. In addition, we compared cortical responses to form and motion stimuli by a receptive-field-based backprojection of cortical activities. RESULTS: We confirmed our previous finding that electrical retina stimulation may yield a spatial resolution of 1-5 degrees visual angle and a temporal resolution of about 20 ms. We found that the spatio temporal cortical activation profiles are commonly related to retinal form and motion stimuli. Cortical activity analyses showed that for two-point form stimuli the neuronal interaction depends on the stimulation electrodes' distance and that local cortical group activities can exhibit some tuning to the directions or the velocities of moving electrical bars'. Projections of cortical activations to visual space were consistent with electrical form and motion stimulation of the retina. CONCLUSIONS: Our data indicate that retinal stimulation with electrical form and motion stimuli can lead to spatio-temporally related cortical activations. However, the selective activation of single cortical neurones with specific visual tuning properties by electrical retina stimulation and the potential adaptation of the visual system to long-term stimulation with retina implants should be addressed in future work. PMID- 12898283 TI - Significance of Aspergillus fumigatus isolation from respiratory specimens of nongranulocytopenic patients. AB - The aim of this study was to determine the significance of isolation of Aspergillus fumigatus from cultures of respiratory specimens in nongranulocytopenic patients. The medical records of patients with respiratory specimens positive for Aspergillus fumigatus who were admitted to an adult pneumology ward were reviewed during a 2-year period. A total of 80 respiratory specimens from 76 patients yielded Aspergillus fumigatus. Forty-eight patients were colonized with Aspergillus fumigatus, whereas the 28 (37%) remaining patients had pulmonary aspergillosis, manifest as aspergilloma ( n=19 patients), chronic necrotizing pulmonary aspergillosis ( n=7 patients), and bronchial aspergillosis ( n=2 patients). The presence of typical hyphae in direct examination of bronchoscopic specimens was more likely to be found in infected than in colonized patients ( P=0.04). No immunological test was positive in colonized patients, whereas anti- Aspergillus antibodies were detected in 55% of infected patients ( P<0.001). Pulmonary tuberculosis was the most common underlying lung disease in patients with aspergilloma, but it was not found in any patient with chronic necrotizing pulmonary aspergillosis ( P=0.006). Anti- Aspergillus antibodies were more likely to be detected in patients with aspergilloma (78%) than in patients with chronic necrotizing pulmonary aspergillosis (14%) ( P=0.007). The analysis of predisposing factors, in conjunction with immunological tests and examination of bronchoscopic specimens, is helpful in distinguishing between colonization and infection with Aspergillus fumigatus, as well as for differentiating between aspergilloma and chronic necrotizing pulmonary aspergillosis. PMID- 12898284 TI - Evaluation of the Merlin, Micronaut system for automated antimicrobial susceptibility testing of Pseudomonas aeruginosa and Burkholderia species isolated from cystic fibrosis patients. AB - Since accurate antimicrobial susceptibility testing of bacterial cystic fibrosis isolates is known to be problematic and an optimal in vitro testing method has not yet been evaluated, the study presented here was conducted to compare the performance of the reference agar dilution method and broth microdilution with a commercially available automated susceptibility test system (Merlin; Micronaut, Germany). In this pilot study, the susceptibility of 70 clinical strains of Pseudomonas aeruginosa and Burkholderia cepacia-like organisms to nine antimicrobial agents was tested using these methods. Susceptibility results generated by broth microdilution (both automated and according to the National Committee for Clinical Laboratory Standards recommendations) were demonstrated to be of good reproducibility, and they compared favourably to the time- and material-consuming standard agar dilution reference method, especially after a prolonged incubation time (48 h). PMID- 12898285 TI - Cultural innovation and transmission of tool use in wild chimpanzees: evidence from field experiments. AB - Chimpanzees (Pan troglodytes) are the most proficient and versatile users of tools in the wild. How such skills become integrated into the behavioural repertoire of wild chimpanzee communities is investigated here by drawing together evidence from three complementary approaches in a group of oil-palm nut- ( Elaeis guineensis) cracking chimpanzees at Bossou, Guinea. First, extensive surveys of communities adjacent to Bossou have shown that population-specific details of tool use, such as the selection of species of nuts as targets for cracking, cannot be explained purely on the basis of ecological differences. Second, a 16-year longitudinal record tracing the development of nut-cracking in individual chimpanzees has highlighted the importance of a critical period for learning (3-5 years of age), while the similar learning contexts experienced by siblings have been found to result in near-perfect (13 out of 14 dyads) inter sibling correspondence in laterality. Third, novel data from field experiments involving the introduction of unfamiliar species of nuts to the Bossou group illuminates key aspects of both cultural innovation and transmission. We show that responses of individuals toward the novel items differ markedly with age, with juveniles being the most likely to explore. Furthermore, subjects are highly specific in their selection of conspecifics as models for observation, attending to the nut-cracking activities of individuals in the same age group or older, but not younger than themselves. Together with the phenomenon of inter-community migration, these results demonstrate a mechanism for the emergence of culture in wild chimpanzees. PMID- 12898286 TI - Dementia with Lewy bodies: disease concept and genetics. AB - Dementia with Lewy bodies (DLB) was first recognized as a clinicopathological entity about 20 years ago. It is the second most-common degenerative dementia after Alzheimer's disease. Clinically, DLB differs from Alzheimer's disease in that disease symptoms are prone to fluctuate and patients often suffer from visual hallucinations, while short-term memory is relatively preserved. As many as 70% of patients have parkinsonism and up to 50% are sensitive to the extrapyramidal side effects of neuroleptic drugs. About 3 million Europeans will be affected by DLB in 2020 if no cure or effective treatment is found. This article reviews the current disease concept, as well as existing problems concerning classification and delineation of DLB from other conditions with dementia. The literature on genetic findings in this complex disease is critically discussed. PMID- 12898287 TI - Association of late-onset Alzheimer disease with a genotype of PLAU, the gene encoding urokinase-type plasminogen activator on chromosome 10q22.2. AB - Urokinase-type plasminogen activator (uPA) converts plasminogen to plasmin. Plasmin is involved in processing of amyloid precursor protein and degrades secreted and aggregated amyloid-beta, a hallmark of Alzheimer disease (AD). PLAU, the gene encoding uPA, maps to chromosome 10q22.2 between two regions showing linkage to late-onset AD (LOAD). We genotyped a frequent C/T single nucleotide polymorphism in codon 141 of PLAU (P141L) in 347 patients with LOAD and 291 control subjects. LOAD was associated with homozygous C/C PLAU genotype in the whole sample (chi2=15.7, P=0.00039, df 2), as well as in all sub-samples stratified by gender or APOE epsilon4 carrier status (chi2> or = 6.84, P< or =0.033, df 2). Odds ratio for LOAD due to homozygosity C/C was 1.89 (95% confidence interval 1.37-2.61). PLAU is a promising new candidate gene for LOAD, with allele C (P141) being a recessive risk allele or allele T (L141) conferring protection. PMID- 12898288 TI - DNA polymorphisms of lipase related genes. AB - We previously determined sequenced the PNLIP gene encoding pancreatic lipase in cell lines of subjects with clinical deficiency of pancreatic lipase (MIM 246600) and found no putative disease-causing mutations. As part of the ongoing analysis of the genomic DNA of these subjects, we now report the development of genomic amplification primers to sequence the coding regions of CLPS, CEL, PLRP1, and PLRP2, encoding pancreatic co-lipase, carboxyl-ester lipase, and pancreatic lipase-related proteins-1 and -2, respectively. Whereas we found no putative disease-causing missense or nonsense mutations in these samples, we discovered a total of 13 common polymorphisms (12 single nucleotide polymorphisms) in these four genes. Genotypes of these polymorphisms may be useful in future association analyses. PMID- 12898289 TI - Linkage and association of the CA repeat polymorphism of the IL6 gene, obesity related phenotypes, and bone mineral density (BMD) in two independent Caucasian populations. AB - Genetic factors play an important role in osteoporosis and obesity, two serious public health problems in the world. We investigated the relationships between obesity-related phenotypes, bone mineral density (BMD) and the CA repeat polymorphism of the IL6 gene in two large independent samples using the quantitative transmission disequilibrium test (QTDT). The first sample consisted of 1,816 individuals from 79 multigenerational pedigrees. Each pedigree was identified through a proband with BMD Z-scores 0.1) at 6-9 years follow-up. More patients had chronic groin pain following Lichtenstein repair (38%) than after Shouldice repair (7%) ( P<0.05). More information is needed on long-term groin pain following anterior mesh repair. The Shouldice inguinal hernia repair may have a role in open primary herniorrhaphy to decrease the risk of chronic groin pain. PMID- 12898291 TI - Analysis of the effect of electrostatic energy truncation in molecular dynamics simulations of immunoglobulin G light chain dimer. AB - Molecular dynamics (MD) simulations of immunoglobulin G (IgG) light chain dimer using particle mesh Ewald (PME) and cutoff methods of treating electrostatic interactions were performed. The results indicate that structural parameters (RMSD, radius of gyration, solvent accessible surface) are very similar for both schemes; however, PME simulation shows increased mobility of side chains. This leads to larger fluctuations in the distance between the monomers in the dimer molecule, and, as a consequence, results in decreased number of interactions across the dimer interface. The wall clock time of the simulations was also compared. It was shown that the PME method is approximately 30% faster than the cutoff method for the system studied on a single processor. PMID- 12898292 TI - Molecular dynamics exposes alpha-helices in myelin basic protein. AB - Molecular dynamics simulations of models of unmodified and deiminated MBP (myelin basic protein) have been performed on solvated structures with added counterions, for 10 ns using AMBER (assisted model building with energy refinement). The protein structures became extended, and a considerable number of alpha-helical segments formed spontaneously. The degree of molecular extension was greater in the deiminated species, and the alpha-helices were more transient. These structural disruptions may be operative in vivo during multiple sclerosis. PMID- 12898293 TI - Influence of salivary pellicle formation time on enamel demineralization--an in situ pilot study. AB - This study assessed the protective potential of salivary pellicles formed in situ over periods ranging from 2 to 24 h. Pellicles were produced on enamel slabs mounted on the palatal aspect of removable acrylic splints and exposed to the oral environment in three subjects for 2, 6, 12 and 24 h. Enamel specimens with and without pellicles were immersed in citric acid (1%) for 60 s, and the amount of dissolved calcium was measured by atomic absorption spectroscopy. In addition, specimens were processed for transmission electron microscopy (TEM). Mean values (standard deviations) for calcium release (mg/l related to the specimen's surface area of 5 x 5 mm(2)) were: 2-h pellicle 6.94 (1.55); 6-h pellicle 6.69 (2.05); 12 h pellicle 6.57 (2.31); 24-h pellicle 5.71 (2.46); enamel without pellicle 8.95 (1.66). There were no significant differences in calcium release that were dependent on pellicle formation time, but in comparison to enamel specimens without pellicle, significantly less (p <0.05) demineralization of the enamel was observed in pellicle-covered specimens. TEM showed that the pellicle was partly, but not completely dissolved following acid exposure. It is concluded that even a 2-h in-situ-formed pellicle layer protects the enamel surface to a certain extent against demineralization. PMID- 12898294 TI - Two-year clinical performance of a packable posterior composite with and without a flowable composite liner. AB - The aim of the study was to evaluate the clinical performance of a packable fine hybrid dental composite (Prodigy Condensable) and the influence of the additional application of a flowable resin composite (Revolution, SDS Kerr) layer on marginal integrity after 2 years in stress-bearing posterior cavities according to the Ryge criteria. In 50 patients (40.5+/-17.5 years of age), 116 class II fillings (metal matrix system, glass ionomer-cement-base in 36%, rubberdam isolation in 70%) were placed, with at least two restorations per patient. The adhesive Optibond Solo Plus was used for all the restorations. In one of the two fillings in each patient, an additional layer of the flowable composite Revolution was applied in the entire cavity and separately light-cured. Baseline scores have been rated Alfa in > or =95% and Bravo in <5%. After 2 years, the results [%] of the Ryge evaluation for the two groups with/without the additional use of Revolution were: (1) Marginal Adaptation: Alfa:78/70, Bravo:16/27, Charlie:0/0, Delta:6/4; (2) Anatomic Form: Alfa:89/95, Bravo:6/2, Charlie:6/4; (3) Secondary Caries: Alfa:98/100, Bravo:2/0; (4) Marginal Discoloration: Alfa:76/68, Bravo:24/32, Charlie:0/0; (5) Surface: Alfa:90/91, Bravo:4/5, Charlie:0/0, Delta:6/4; (6) Color Match: Oscar:56/57, Alfa:44/39, Bravo:0/4, Charlie:0/0. Within the observation period (recall rate: 95%), three restorations out of 116 at baseline fractured, one restoration showed a secondary caries, one tooth received endodontic treatment, and all other restored teeth remained vital. After 2 years, no statistically significant difference (Chi-square test) in the overall survival rate between the group with the additional use of Revolution (92.8%) and that without Revolution (94.6%) was found. The combined survival rate for both groups together was 93.7% of clinically acceptable restorations. PMID- 12898295 TI - Magnetic resonance imaging diagnosis and new classification of the osteoporotic vertebral fracture. AB - The purpose of this study was to investigate the usefulness of magnetic resonance imaging (MRI) in the diagnosis of, and when deciding on treatment policies for, vertebral fractures in patients with osteoporosis by comparing the diagnostic success rates for such fractures using MRI and plain radiographs. We also devised a new classification of vertebral fractures in osteoporotic patients based on MRI findings. The subjects were 34 patients with a total of 316 osteoporosis-related vertebral fractures treated in our department during the past 2 years. They consisted of 30 women and 4 men with an average age of 82 years (range 61-95 years). The site diagnostic rates of plain radiographs and MRI for the vertebral fractures were compared, and changes in signal intensity of the fractured vertebrae in MRI were examined. The areas of change in signal intensity in T1 weighted sagittal images were classified into six types: total, anterior, posterior, superior, inferior, central. The site diagnostic rate for vertebral fractures was 98% for MRI, which was better than the 87% obtained with plain radiographs. Changes in signal intensity in the vertebrae showed no intraspinal protrusion in the inferior and superior types, and neurological stability was achieved; but there was a high frequency of intraspinal protrusion in the total and posterior types. PMID- 12898296 TI - Quantitative assessment of hand radiographs of rheumatoid arthritis: interobserver variation in a multicenter radiographic study. AB - We investigated interobserver variations in the Larsen radiographic scoring method on hand radiographs of rheumatoid arthritis (RA) patients in a multicenter trial and developed a new radiographic scoring method. Thirteen experienced rheumatologists scored 10 representative RA hand radiograms with the Larsen scoring method and clarified the precipitating factors of interobserver variation. Based on this study, it was proved that the ankylotic joint, overlapping joint, and more precise erosive joint are needed for optimal radiographic evaluation. Therefore, we modified the Larsen scoring method on the basis of these precipitating factors and developed a novel radiographic scoring method. Finally, to determine which scoring system was most reliable, the interobserver variation using three methods (original Larsen method, revised Larsen method, our scoring method) were compared by 13 experienced rheumatologists and 13 residents. Our scoring method proved to have simplicity, reliability, and ease of learning. These results suggest that our novel radiological quantitative assessment method has useful applications for clinical studies in patients with RA. PMID- 12898297 TI - Effect of extracorporeal shock waves on callus formation during bone lengthening. AB - The effects of extracorporeal shock waves (ESWs) on callus formation during bone lengthening were studied in 25 female Japanese white rabbits. Bone lengthening of 9.8 mm was obtained over 2 weeks using the Orthofix M-100 bone fixator. ESWs were applied 3 weeks after surgery. Pins were removed 7 weeks after surgery, and specimens were prepared after the animals were killed at 9 and 24 weeks. The shock wave setting used was 0.42 mJ/mm(2) with a pulse interval of 2 Hz; 3000 shots each were applied to the central and peripheral areas. The specimens were evaluated using radiography, bone mineral density (BMD) measured by dual energy absorptiometry (DXA), and a three-point bending test to evaluate mechanical strength. Histological examination was performed on the lengthened portion. Radiographs and histological observations revealed no apparent fractures in nonlengthening tibias at the shock wave energy densities used. Radiographic observations revealed no apparent differences between the control group and the ESW group. BMD measurements by DXA revealed significantly increased bone mass in the ESW group 9 weeks after surgery. At 24 weeks after surgery the mean BMD had decreased to 25% and 15% of the values at 9 weeks in the control and ESW groups, respectively. The three-point bending test revealed no significant differences between the groups. Histological observations revealed significant capillary formation and osteoblasts and chondrocytes in the bone marrow as well as bridging of newly formed trabeculae 2 weeks after the bone was lengthened. At 4 weeks after treatment, observations included parts of the lengthened portion with no cortex or immature bone. At 9 weeks after surgery, cortex formation and a normal medullary cavity were clearly observed in the control group, whereas observations in the treated group included areas of the lengthened portion with no cortex and formation of immature trabecular structures and increased cancellous bone in the center of the lengthened portion. At 24 weeks after surgery, more prominent cortex formation and fatty marrow were observed in the ESW group than in the control group. PMID- 12898298 TI - Two-dimensional gel electrophoresis of synovial fluid: method for detecting candidate protein markers for osteoarthritis. AB - Synovial fluid (SF) is a dynamic reservoir for proteins originating from serum, synovial tissue, and cartilage. The composition of the SF proteome may reflect the pathophysiological conditions affecting the circulatory system and cartilage. Our long-term goal is to identify reliable protein markers for osteoarthritis (OA) in SF. We first evaluated the pattern of SF proteins on two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) as a function of protein loading, pH range for isoelectric focusing, and concentration of acrylamide in SDS-PAGE. Removal of albumin and Gamma-globulins from the samples did not improve the detection of protein spots on 2D-PAGE. The repeatability of protein spot intensity was tested by triplicate 2D-PAGE of a given sample; these experiments showed low intrasample variability (correlation coefficients 0.89-0.95). Differences between multiple samples were tested by comparing the 2D-PAGE of four samples. These experiments showed slightly greater variation between samples (correlation coefficients 0.85-0.93) and a number of differentially expressed proteins. The intensity of 18 protein spots differed more than fivefold, and the intensity of nine protein spots differed more than 100-fold. These results show that 2D-PAGE can be used under standard conditions to screen SF samples and identify a small subset of proteins in SF that are potential markers associated with OA. PMID- 12898299 TI - Validation of acetabular cup wear volume based on direct and two-dimensional measurements: hip simulator analysis. AB - The volumetric wear in retrieved cups can be assessed by mathematical conversion based on linear measurements and by a fluid-displacement method. We used a hip simulator model to produce wear in 22-, 28-, and 32-mm hip implants and then assessed the volumetric wear using a gravimetric wear method. We then compared the findings with those obtained with the linear and fluid-displacement methods. For the linear method, we translated the linear wear to the volumetric wear using the equations developed by Charnley et al., Kabo et al., and Hashimoto et al. The fluid-displacement method showed the strongest correlation with the gravimetric wear method, and it was found to overestimate the volume slightly (by 3%-9%). According to the linear wear conversion, however, the equation by Kabo widely underestimated the volume by 33%-40%. The equation used by Charnley tended to overestimate the volume (by 4%-17%), whereas Hashimoto's equation tended to slightly underestimate the volume (by 2%-12%). The fluid-displacement method demonstrated an average error of 0.34% +/- 13.40% when the wear exceeded 400 mm(3). The linear wear was thus converted to the volume wear most accurately using Hashimoto's equation, with the average error being -3.8% +/- 14.0%. Of the four measurement modalities, the fluid-displacement method showed the most accurate results. We therefore confirmed that the fluid-displacement method is the most accurate way to determine volumetric wear in retrieved cups. PMID- 12898300 TI - Evaluation of sensory function after median nerve decompression in carpal tunnel syndrome using the current perception threshold test. AB - The recovery level for sensory function after carpal tunnel release for the treatment of idiopathic carpal tunnel syndrome (CTS) was assessed with the current perception threshold (CPT) test. Seventeen CTS patients (21 hands) were followed, and the CPTs at the index finger of each patient was measured preoperatively and at 1, 3, and 6 months postoperatively. After carpal tunnel release, there was significant recovery of CPT at all stimulation frequencies, indicating improvement of all sensory functions including sensations of temperature, pain, touch, and vibration. PMID- 12898301 TI - Biomechanical analysis for stress fractures of the anterior middle third of the tibia in athletes: nonlinear analysis using a three-dimensional finite element method. AB - We evaluated stresses in the anterior middle third of the tibia that have been reported to predict a poor prognosis for tibial stress fractures compared to other predominant sites (posteromedial regions of the distal third and proximal third). The effect of two different loads (bending-compression load and torsional load) on three sites was investigated using a three-dimensional finite element method. The model was constructed using the tibia, fibula, proximal tibiofibular joint, interosseous membrane, and tibiofibular ligament based on computed tomography scans obtained at 4-mm intervals of the lower leg of a 20-year-old woman who exhibited no abnormal findings on roentgenograms. First, a normal model was constructed using normal material properties, and then the model was modified to produce fracture models by varying the mechanical properties of each predominant site and expanding the area in three gradual phases on the assumption that the fracture advanced in three phases. Each model was tested against the same two loads, and stresses at the nodal points on the border of the fracture area and normal area were compared in each cross section to determine the effect of the load on fracture advancement. In response to torsional load, both the normal model and fracture models tended to show higher values for the posteromedial distal third than the anterior middle third. By examining the bending-compression load it could be seen that the mean peak value significantly decreased between the first and second phases in fracture models of the anterior middle third. This finding was inconsistent with our previous belief that the bending-compression load would have more serious consequences than the torsional load. In contrast, when the area of fracture was expanded into the third phase, maximum values were significantly higher than during the second phase. No similar finding was observed for the posteromedial distal third, suggesting that the anterior middle third may have the same stable biomechanical conditions as the posteromedial distal third at an earlier stage and thus have little influence on fractures. When the fracture is more advanced, however, the conditions change suddenly, and a bending-compression load may adversely affect the mechanical conditions in this area and thereby cause complete fracture. PMID- 12898302 TI - Evaluation of articular cartilage with 3D-SPGR MRI after autologous chondrocyte implantation. AB - To assess the maturation process of the cartilage after autologous chondrocyte implantation (ACI), we performed a longitudinal study with three-dimensional spoiled gradient-recalled (3D-SPGR) magnetic resonance imaging (MRI). Five knees of five patients on which ACI of the femoral condyle was performed were studied. The signal intensity of reparative tissue approached that of normal articular cartilage with time. The volume of reparative tissue remained at an almost constant level after implantation. During second-look arthroscopy, the areas on which ACI was performed were covered with hyaline-like cartilage, and the reparative tissue removed by biopsy consisted of normal chondrocytes and extracellular matrix. The increased signal intensity of the reparative tissue represents maturation of implanted autologous chondrocytes. 3D-SPGR MRI is thought to be useful for evaluating reparative tissue after autologous chondrocyte implantation. PMID- 12898303 TI - Simultaneous live video presentation during knee arthroscopy. AB - To improve patient understanding, we evaluated prospectively the effect of patient-observed simultaneous surgeon-explained live video during knee arthroscopy. Patient satisfaction, understanding, and tolerance of the tenderness and pain were measured by a questionnaire completed immediately after the procedure. There were 72 patients (41 men, 31 women) with a mean age of 47 years; 76 joints were treated among them. All but one were comfortably able to watch the live video of the procedure, and 89% of the patients reported a good understanding of and satisfaction with the procedure. Moreover, if the patients ever had to have another arthroscopy, 66% answered that they "certainly" wanted a live video presentation, and 25% "somewhat" desired it. Live video-presented outpatient arthroscopic knee surgery could be considered a method for improving patient understanding of knee pathology. PMID- 12898304 TI - Medial approach without opening the joint capsule for developmental dislocation of the hip. AB - Seventy-one developmentally dislocated hips in 47 children with an average age of 11 months (range 4-16 months) were treated by the medial approach without opening the capsule followed by 6 months of hip spica-cast immobilization. At the latest follow-up 63 hips were rated excellent and good, and 8 hips showed subluxation. No avascular necrosis had occurred. It is concluded that this approach is safe and effective for treating developmental dislocation of the hip. Moreover, opening the capsule is not only an unnecessary task in this age group but also should be avoided to prevent avascular necrosis. PMID- 12898305 TI - Changes in anteroposterior stability following total knee arthroplasty. AB - This study evaluated the changes in anteroposterior stability at flexion and extension following posterior cruciate ligament (PCL)-retaining and PCL substituting total knee arthroplasty (TKA), in comparison with normal knees. Altogether, 29 knees of 24 patients underwent either PCL-retaining ( n = 19) or PCL-substituting ( n = 10) TKA. Anteroposterior displacement was measured with a KT-2000 arthrometer preoperatively, 1 month after surgery, and again an average of 3 years after surgery. Anteroposterior stability at flexion and extension did not change following PCL-retaining TKA and was not significantly different from that of normal subjects, whereas anteroposterior stability increased following PCL-substituting TKA and was significantly greater than that of normal subjects an average of 3 years after surgery. PMID- 12898306 TI - Effects of cyclical etidronate with alfacalcidol on lumbar bone mineral density, bone resorption, and back pain in postmenopausal women with osteoporosis. AB - The purpose of the present open-labeled, randomized, prospective study was to compare the effects of cyclical etidronate combined with alfacalcidol with those of cyclical etidronate alone on lumbar bone mineral density (BMD), bone resorption, and back pain in postmenopausal women with osteoporosis. Forty postmenopausal women with osteoporosis, 60-86 years of age, without any vertebral fractures in the lumbar spine, were randomly divided into two groups with 20 patients in each group. One group was treated with cyclical etidronate (oral etidronate 200 mg daily for 2 weeks every 3 months) and the other was given cyclical etidronate combined with alfacalcidol (cyclical etidronate plus alfacalcidol 1 Ig daily continuously). The BMD of the lumbar spine (L1-L4) measured by dual-energy X-ray absorptiometry, urinary crosslinked N-terminal telopeptides of type I collagen (NTX) measured by an enzyme-linked immunosorbent assay, and back pain evaluated by the face scale score were assessed at baseline, 6 months, and 12 months. There were no significant differences in baseline characteristics including age, body mass index, years since menopause, lumbar BMD, urinary NTX level, and face scale score between the two treatment groups. Both treatments significantly reduced the urinary NTX level and back pain. Cyclical etidronate combined with alfacalcidol significantly increased the lumbar BMD with a more significant reduction in the urinary NTX level than cyclical etidronate alone, but cyclical etidronate alone did not significantly increase the lumbar BMD. Alleviation of back pain was similar in the two groups. These results suggest that cyclical etidronate combined with alfacalcidol appears to be more useful than cyclical etidronate alone for increasing the lumbar BMD by more markedly suppressing bone resorption in postmenopausal women with osteoporosis. PMID- 12898307 TI - Coronal laxity in extension in vivo after total knee arthroplasty. AB - We performed stress arthrometric studies on 77 knees (71 patients) with total knee arthroplasty to determine the presence and magnitude of femoral abduction and adduction in knee extension. A total of 53 knees (49 patients) had posterior cruciate ligament-retaining (PCLR) prostheses, and 24 (22 patients) had PCL substituting (PCLS) prostheses. The selected patients had successful arthroplasties with no clinical complications a minimum of 5 years after primary surgery. Each patient was subjected to a successive abduction and adduction stress test at 0 degrees -20 degrees of flexion using a Telos arthrometer. The mean values for abduction and adduction were 4.8 degrees and 4.5 degrees with a PCLR prosthesis, respectively, and 4.6 degrees and 4.0 degrees with a PCLS prosthesis. There were no statistical differences between PCLR and PCLS knees. The results suggest that approximately 4 degrees of laxity in these directions is suitable in total knee arthroplasty for a satisfactory clinical outcome 5-9 years after surgery. PMID- 12898308 TI - Validation of the Japanese version of the Roland-Morris Disability Questionnaire. AB - The study was designed to validate a translated, culturally adapted questionnaire. We examined the reliability, validity, and responsiveness of the Japanese version of the Roland-Morris Questionnaire (RDQ) when assessing disability in Japanese patients with low back pain. The RDQ is a reliable, validated scale used to measure disability caused by low back pain. However, no validated Japanese version of this questionnaire is available. A series of 214 outpatients with low back pain participated in this validation study. The patients were given the RDQ and the SF-36, and assessed their pain and global rating of health. Among them, 57 who were clinically stable were given the RDQ again 2 weeks later. The reliability was examined based on the test-retest method and internal consistency. Sufficient reliability was demonstrated with a Chronbach's. coefficient of 0.85, and the reproducibility for the 30 patients was r = 0.91. The principal component analysis showed unidimensionality. The RDQ score of the 133 patients was significantly improved after treatment. The Japanese version of the RDQ is a useful scale that is easy to use with reliability, validity, and responsiveness when assessing patients with low back pain. PMID- 12898309 TI - Tissue response to porous hydroxyapatite ceramic in the human femoral head. AB - We describe pathohistological findings of the human femoral head after it was grafted with hydroxyapatite (HA) and allogenic bone. The femoral head was removed because of the recurrence of a giant cell tumor 15 months after the graft. Histological investigation revealed that the entire surface of the HA granules was completely surrounded by the bone in the periphery of the grafted area; in some areas granules were partly in contact with the existing bone, and the rest were surrounded by fibrous tissue. There was no intervening layer of fibrous tissue between the granule surface and the bone where they were in direct contact. Pores were completely filled with fibrous tissue, partially filled with bone in the periphery of the pores, or completely filled with bone. Undecalcified histology revealed the formation of osteoid and ossification at the site where fibrous tissue infiltrated the pores. These findings indicated continued ingrowth of new bone into the HA pores. Under a scanning electron microscope, the surfaces of the granules showed clear demarcation in vitro, but it became less smooth, indicating in vivo changes. Although no foreign body reaction to the HA was noted anywhere, the HA surface appeared to be subject to a biodegradation process. PMID- 12898310 TI - Recognition by cellular and humoral autologous immunity in a human osteosarcoma cell line. AB - Because of the difficulty of developing pairs of osteosarcoma cell lines and cytotoxic T lymphocytes (CTLs), no osteosarcoma tumor antigens that are useful for antiosteosarcoma immunotherapy have yet been identified. In parallel with continuous attempts to develop such pairs from osteosarcoma, we employed serological identification using a recombinant expression cloning (SEREX) method to identify B cell-defined antigens. Consequently, a human osteosarcoma cell line, OS2000, was established from a primary osteosarcoma of a patient cured of hereditary retinoblastoma. Repetitious in vitro stimulations by OS2000 cells to the autologous peripheral T cells induced cytotoxic activity in the autologous osteosarcoma cells but not in the nontumor cells. The cytotoxicity was inhibited by anti-HLA class I monoclonal antibody. SEREX analysis revealed that autologous humoral immunity reacted to two proteins expressed in OS2000. One was the self HLA-Cw*0102 molecule, and the other was wild-type smooth muscle myosin light chain (SMMLC). However, no antigenicity of these proteins was seen versus the sera of the other patients. In conclusion, our results demonstrated the presence of host cellular and humoral immune responses to autologous osteosarcoma cells. This offered the opportunity to identify osteosarcoma antigens recognized by autologous immunity. PMID- 12898311 TI - Experimental evaluation of the usefulness of osteochondral allograft for articular cartilage defect. AB - This study was conducted to evaluate the usefulness of osteochondral allografts for articular cartilage defects. Cartilaginous defects measuring 4.5 mm in diameter were experimentally prepared in both the weight-bearing and non-weight bearing regions of the femur in six male miniature pigs (9 months old). Osteochondral grafting was performed using fresh autografts (group AU), fresh allografts (group AL), or frozen allografts (group FA). Untreated cartilaginous defects were used as the control (group D). All the pigs were killed 4 weeks later, and the respective osteochondral grafts were macro- and microscopically evaluated. Hematoxylin and eosin staining, safranin O staining, and immunostaining [matrix metalloprotease-1 (MMP-1) and tissue inhibitor of metalloprotease-2 (TIMP-2)] were used for the histological evaluations of transplanted cartilage. Macroscopic and microscopic findings were assessed according to the criteria proposed by Wakitani et al. Although groups AU and AL showed similar median scores (ranges) for the evaluation of cartilaginous defect restoration, groups FA and D showed unfavorable scores: 3.9 (0-9) in group AU; 4.5 (0-12) in group AL; 10.2 (4-12) in group FA; and 7.0 (5-11) in group D. Immunostaining revealed almost identical results in groups AU and AL. As there were no histologically significant differences in the status of the osteochondral grafts between fresh autografts and fresh allografts, fresh allografts might be useful donor osteochondral grafts. PMID- 12898312 TI - Effects of anterior lumbar spinal fusion on the distribution of nerve endings and mechanoreceptors in the rabbit facet joint: quantitative histological analysis. AB - The purpose of this study was to investigate the density and distribution of neural endings in rabbit lumbar facet joints after anterior spinal fusion and to evaluate the effects of intervertebral immobilization. An extraperitoneal approach was applied, and L5/6 was fixed with a plate and screws. Bilateral L4/5, L5/6, and L6/7 facet joint capsules were harvested from the rabbits 4, 8, and 16 weeks postoperatively. Capsular tissues were processed using a modified gold chloride staining method, and the specimens were sliced into 15-micro m sections. All sections were analyzed microscopically, and neural ending numbers per unit volume were calculated. Three types of neural ending were identified in each specimen: Pacinian corpuscles, Ruffini corpuscles, and free nerve endings. In the L5/6 fusion segment there was a significant decrease in the number of Pacinian corpuscles at 4 weeks and of Ruffini corpuscles at 4, 8, and 16 weeks after the fusion compared with the control; and in the L4/5 upper adjacent segment there was a significant increase in the number of free nerve endings. The number of Ruffini endings for the L6/7 lower adjacent segment was significantly lower more than 8 weeks after the fusion. These results suggest that immobilization of the intervertebral segment causes a reduction in the number of mechanoreceptors in the facet joint capsules because of the reduction in mechanical stimulation. Moreover, in the upper adjacent facet joint there may be neural sprouting caused by nociceptive stimulation. PMID- 12898313 TI - Effects of lumbar spinal fusion on the other lumbar intervertebral levels (three dimensional finite element analysis). AB - The risk of accelerating the degeneration of adjacent disc levels after lumbar spinal fusion is a controversial issue. A finite element model consisting of L1 to L5 lumbar spines was used to assess the effect on adjacent disc level after lumbar spinal fusion. We compared intact, L4/5 posterior interbody fusion (PLF), and L4/5 posterior lumbar interbody fusion (PLIF) models. The loading conditions applied were compressive force, compressive force plus flexion moment, and compressive force plus extension moment. Evaluations were made for von Mises stress on each vertebral end-plate, Tresca stress of all the annulus fibrosus, and Tresca stress of the annulus fibrosus from the posterior surface of the disc to the neural foramen. As the result, the von Mises stress adjacent to the fusion level was higher than the other nonfusion levels; it was higher under conditions of flexion moment loading plus compression loading [112% (2.59 PMa) in the PLF model and 117% (2.72 Mpa) in the PLIF model] than in the intact model. The Tresca stress of all the annulus fibrosus adjacent to the fusion level was higher than that on other nonfusion intervertebral levels; it was higher under conditions of flexion moment loading plus compression loading [127% (0.57 PMa) in the PLF model and 209% (0.89 Mpa) in the PLIF model] than in the intact model. The Tresca stress of the annulus fibrosus from the posterior surface of the disc to the neural foramen adjacent to the fusion level was higher than that on other nonfusion intervertebral levels; and it was higher under conditions of flexion moment loading plus compression loading [107% (1.48 PMa) in the PLF model and 112% (1.54 Mpa) in the PLIF model] than in the intact model. These findings demonstrate that with lumbar fusion, stresses on the vertebral end-plate and the annulus fibrosus were high adjacent to the fusion level; furthermore, stresses were higher in the PLIF model than in the PLF model. These results suggested that lumbar spinal fusion might bring with it a risk of damage to the annulus fibrosus and the vertebral end-plate adjacent to the fusion level. PMID- 12898314 TI - Determination of the complete cDNA sequence of rat type II collagen and evaluation of distinct expression patterns of types IIA and IIB procollagen mRNAs during fracture repair in rats. AB - Elucidating the molecular mechanisms that underlie fracture healing is crucial to understanding and devising strategies for the management of fractures, especially those associated with a pathological condition such as diabetes or old age. Cartilage formation, and therefore the expression of type II collagen by chondrocytes, is a critical step in frac-ture healing. Two forms of type II collagen, IIA and IIB, are known to be produced by alternative splicing of the Alpha(1) (II) procollagen gene. We have followed the patterns of expression of these two forms of type II collagen to determine the nature of chondrocyte recruitment during fracture healing. First, we sequenced the rat collagen type II cDNA to design the primers. Second, using a competitive quantitative reverse transcription-mediated polymerase chain reaction, we provide evidence that (1) there is a basal level of type IIA collagen expression during the early stages of fracture healing; (2) transient but sharp up-regulation of IIA expression occurs concomitant with chondrogenesis and endochondral ossification; and (3) type IIB collagen is the predominant mRNA variant expressed at virtually all times during fracture repair. PMID- 12898315 TI - Synovial fluid depletion: successful arthrodesis without operative cartilage removal. AB - Operative arthrodesis has been thought to require surgical removal of remaining joint cartilage, but we have found in rabbits that even a joint with intact cartilage can fuse if rigidly fixed. This may enable new percutaneous techniques for arthrodesis. Rigid adaptation of the joint surfaces deprives the cartilage of mechanical stimulation and depletes the cells of synovial fluid transport of oxygen and nutrition. To better understand the requirements for successful arthrodesis, we studied the histological consequences of the complete absence of mechanical stimulation alone or in combination with synovial depletion by placing a metal cap over part of the joint cartilage in rabbits. The cap was either closed or had an opening to permit synovial fluid to reach the cartilage. We also studied if penetration of the bone-cartilage junction by a drill hole would facilitate cartilage resorption. Synovial fluid depletion in combination with a drill hole through the bone-cartilage junction led to disappearance of all cartilage matrix after 7 weeks. Synovial fluid depletion with an intact bone cartilage junction led to complete disappearance of the cartilage matrix in four of seven rabbits after 7 weeks. With a hole in the cap for synovial fluid, the cartilage matrix was still present to varying degrees after 7 weeks in all the rabbits. In conclusion, percutaneous arthrodesis by rigid adaptation may lead to cartilage disappearance due to synovial depletion rather than due to the absence of mechanical stimulation. A combination with perforation of the bone-cartilage junction appears to lead to reasonably quick removal of the cartilage matrix. PMID- 12898316 TI - Carpal tunnel syndrome due to heterotopic ossification. AB - We report a rare case of carpal tunnel syndrome due to heterotopic ossification in the carpal tunnel in a 34-year-old woman without antecedents of neurological injury, musculoskeletal trauma, or metabolic disorder. To our knowledge, this is the first reported case. Incomplete excision of heterotopic ossification resulted in partial relief of symptoms. PMID- 12898317 TI - Unusually large solitary unicameral bone cyst: case report. AB - A 15-year-old boy presented with complaints of pain and swelling of the right leg. Radiography revealed a large lytic lesion involving the proximal half of the tibia. The patient was taken to surgery, where the cavity was curetted and packed with allogeneic bone graft. Tissue was sent for histopathological evaluation. The patient was kept in an above-knee cast for 4 months, after which partial weight bearing was allowed. The histopathological diagnosis was a unicameral bone cyst. A radiograph at 1 year showed incorporation of the graft, but a small part of the cavity was still visible and there was a persistent discharge. The patient was again subjected to allogeneic bone grafting in the remaining cavity. At 3.5 years of follow-up now, the patient is walking unsupported, there is no pain, and the radiograph shows complete obliteration of the cavity. Unicameral bone cysts are usually a few centimeters in size. To the best of our knowledge, a unicameral bone cyst of such a large size has never been reported in the literature. PMID- 12898318 TI - Incomplete Brown-Sequard syndrome caused by cervical kyphosis secondary to neurofibromatosis: report of a case. AB - The case of a 17-year-old boy who suffered from incomplete Brown-SEquard syndrome caused by dystrophic cervical kyphosis secondary to neurofibromatosis is presented. He became aware of weakness of his left upper and lower extremities and was unable to walk within 1 month of his first feeling numbness in the right upper and lower extremities. Thereafter his condition deteriorated rapidly. On radiological investigation the entire dural sac was shifted anteriorly and the spinal cord was flattened anteroposteriorly and asymmetrically at the angular kyphosis level, as in Hirayama's disease. Despite morphological similarities to Hirayama's disease, this patient presented with quite different clinical features: incomplete Brown-SEquard syndrome and rapidly progressive symptoms. It is suspected that the myelopathy in this case can be attributed to compression of the spinal cord between the vertebral body and the posterior dural wall on only one side. He was successfully treated with posterior interspinous wiring combined with anterior decompression and fusion. The neurological deficit fully disappeared, and complete bony union was obtained 1 year after the operation. PMID- 12898319 TI - Locking of the thumb in a patient with de Quervain's disease: a case report. AB - We report a patient with a locked left thumb in association with de Quervain's disease. While bathing her baby 1 month after giving birth a 32-year-old woman suddenly noticed that she could not radially abduct her left thumb. Magnetic resonance imaging showed thickening of the abductor pollicis longus tendon with a heterogeneous signal intensity on T2-weighted images. Bandage fixation for 4 weeks did not improve her thumb movement, and she was subsequently treated by surgery. Operative findings revealed inhibition of the tendon gliding proximally as a result of nodule formation in the abductor pollicis longus tendon distal to the first dorsal compartment. This condition, locking of the thumb, was improved by excising the extensor retinaculum of the first dorsal compartment and tenosynovium around the abductor pollicis longus and extensor pollicis brevis tendon. At the 1-year follow-up examination the patient had no limitations or pain during active radial abduction of the left thumb. PMID- 12898320 TI - Hip dysplasia in Charcot-Marie-Tooth disease: report of a family. AB - Charcot-Marie-Tooth disease is classified into hereditary motor and sensory neuropathy (HMSN) types I and II, and affected patients present with progressive peripheral neuropathy. Some previous orthopedic studies have revealed the association of hip dysplasia with HMSN, in addition to pes cavovarus, scoliosis, and recurrent dislocation of the patella. We describe three patients from the same family who were each diagnosed as having HMSN type I with associated bilateral severe hip dysplasia, borderline abnormalities of both acetabula, and dysplastic osteoarthritis. Based on our experience with these patients and a review of previous reports, we concluded that routine screening of hip joints, especially for those with a family history of HMSN, is necessary for early diagnosis. PMID- 12898321 TI - Mobile submuscular mass resembling a hard-boiled egg: case report. AB - A 68-year-old woman had a rare mobile mass on her back. The mass was similar to encapsulated fat necrosis except for its unusual stalked appearance, large size, and high degree of mobility underneath the anterior serratus muscle on the back. Based on previous descriptions of encapsulated fat necrosis, the clinical presentation of the mass might be regarded as an extreme example of this rare condition. PMID- 12898322 TI - Unilateral drop finger due to cervical spondylosis at the C6/7 intervertebral level. AB - A 64-year-old man who presented right drop fingers without pyramidal signs due to cervical spondylosis is reported. Magnetic resonance imaging showed a high signal intensity change on T2-weighted scans of the spinal cord at the C6/7 intervertebral level. Evoked spinal cord potentials following ulnar nerve stimulation were attenuated at the C6/7 intervertebral level. Compound muscle action potentials (CMAPs) following right cervical nerve root stimulation were recorded from the extensor digitorum communis. CMAPs following right C8 nerve root stimulation were evoked with small amplitudes. Small polyphasic CMAPs with prolonged onset latency were recorded following right C7 nerve root stimulation. Simultaneous C7 radiculopathy and C8 segmental spinal cord lesion due to cervical spondylosis at the C6/7 intervertebral level were the causes of drop fingers in the present case. Unilateral drop finger is a clinical symptom commonly associated with posterior interosseus nerve palsy, but mention should be made about cervical lesions causing drop finger. Electromyographic abnormalities of the triceps and first dorsal interosseus muscle were the key findings for differentiating the cause from paralysis of the posterior interosseous nerve. PMID- 12898324 TI - The biochemical and biological functions of human papillomavirus type 16 E5 protein. AB - Human papillomavirus type 16 (HPV-16) E5 protein, along with the more publicized E6 and E7 proteins of this virus, has been found to be oncogenic. E5 is a highly hydrophobic membrane-bound protein of 83 amino acids associated with the Golgi apparatus, endoplasmic reticulum, and nuclear membrane in infected cells. E5 can activate epidermal growth factor receptor (EGFR) through binding to the 16 kD subunit of protein pump ATPase leading to a reduced downregulation of EGFR receptors. The activation of EGFR can initiate biochemical cascades that lead to overexpression of a variety of protooncogenes and stimulate rapid cell growth. Moreover, E5 can inhibit the expression of tumor suppressor gene p21((WafI/SdiI/CipI)) and impair the control of cell cycle checkpoint. E5 protein has been identified as a potential tumor vaccine target antigen. PMID- 12898323 TI - Models of the bis-histidine-coordinated ferricytochromes: Mossbauer and EPR spectroscopic studies of low-spin iron(III) tetrapyrroles of various electronic ground states and axial ligand orientations. AB - The EPR and magnetic Mossbauer spectra of a series of axial ligand complexes of tetrakis(2,6-dimethoxyphenyl)porphyrinatoiron(III), [(2,6 (OMe)(2))(4)TPPFeL(2)](+), where L= N-methylimidazole, 2-methylimidazole, or 4 (dimethylamino)pyridine, of one axial ligand complex of tetraphenylporphyrin, the bis(4-cyanopyridine) complex [TPPFe(4-CNPy)(2)](+), and of one axial ligand complex of tetraphenylchlorin, [TPCFe(ImH)(2)](+), where ImH=imidazole, have been investigated and compared to those of low-spin Fe(III) porphyrinates and ferriheme proteins reported in the literature. On the basis of this and previous complementary spectroscopic investigations, three types of complexes have been identified: those having (d(xy))(2)(d(xz),d(yz))(3) electronic ground states with axial ligands aligned in perpendicular planes (Type I), those having (d(xy))(2)(d(xz),d(yz))(3) electronic ground states with axial ligands aligned in parallel planes (Type II), and those having the novel (d(xz),d(yz))(4)(d(xy))(1) electronic ground state (Type III). A subset of the latter type, with planar axial ligands aligned parallel to each other or strong macrocycle asymmetry that yield rhombic EPR spectra, cannot be created using the porphyrinate ligand. Type I centers are characterized by "large g(max)" EPR spectra with g>3.2 and well resolved, widely spread magnetic Mossbauer spectra having A(zz)/ g(N)mu(N)>680 kG, with A(xx) negative in sign but much smaller in magnitude than A(zz), while Type II centers have well-resolved rhombic EPR spectra with g(zz)=2.4-3.1 and also less-resolved magnetic Mossbauer spectra, and usually have A(zz)/ g(Nmu(N) in the range of 440-660 kG (but in certain cases as small as 180 kG) and A(xx) again negative in sign but only somewhat smaller (but occasionally larger in magnitude) than A(zz), and Type III centers have axial EPR spectra with g( upper left and right quadrants ) approximately 2.6 or smaller and g( vertical line )<1.0-1.95, but often not resolved, and less-resolved magnetic Mossbauer spectra having A(zz)/ g(N)mu(N) in the range of 270-400 kG, and A(xx) again negative in sign but much smaller in magnitude than A(zz). An exception to this rule is [TPPFe(4-CNPy)(2)](+), which has A(xx)/ g(N)mu(N)=-565 kG, A(yy)/ g(N)mu(N)=629 kG, and A(zz)/ g(N)mu(N)=4 kG. A subset of Type II complexes (Type II') have rhombicities ( V/Delta) much greater than 0.67 and A(zz)/ g(N)mu(N) ranging from 320 to 170 kG, with A(xx) also negative but with the magnitude of A(xx) significantly larger than that of A(zz). These classifications are also observed for a variety of ferriheme proteins, and they lead to linear correlations between A(zz) and either A(xx), g(zz), or V/Delta for Types I and II (but not for A(zz) versus V/Delta for Type II'). Not enough data are yet available on Type III complexes to determine what, if any, correlations may be observed. PMID- 12898325 TI - Characterisation of a pestivirus isolated from persistently infected mousedeer ( Tragulus javanicus). AB - Serum samples from the male Mousedeer A and the mother, father and sister of A were tested for bovine virus diarrhoea viruses (BVDV) by isolation, and for BVDV antibodies by blocking ELISA and homologous neutralisation test. Further, RNA was extracted and tested by RT-PCR protocol analysing the 5'-untranslated region and the E2 gene of pestivirus. The RT-PCR products were subsequently sequenced. Mousedeer A was positive in virus isolation on three occasions (days 1, 19 and 40) and by RT-PCR. The sister and mother of Mousedeer A were also found virus positive by isolation and RT-PCR. Mousedeer A, its sister and its mother, all had an antibody neutralisation titer below 10. The father of A was virus negative but was positive in the blocking antibody ELISA and had a high neutralisation antibody titer. The repeated detection of BVDV in Mousedeer A, the high amount of virus in serum, the lack of antibodies and the virus positive family members documented that the mousedeer were persistently infected with a pestivirus. The father of A probably had an acute infection resulting in antibodies to pestivirus and viral clearance. Sequence analysis and phylogenetic analysis revealed that the mousedeer pestivirus was closely related to BVDV Type 1f. The existences of persistently infected animals in non-domestic species have great implications for BVDV eradication campaigns in cattle. PMID- 12898326 TI - Changes in level of virus accumulation and incidence of infection are critical in the characterization of Rice tungro bacilliform virus (RTBV) resistance in rice. AB - Analysis by enzyme-linked immunosorbent assay showed that Rice tungro bacilliform virus (RTBV) accumulated in a cyclic pattern from early to late stages of infection in tungro-susceptible variety, Taichung Native 1 (TN1), and resistant variety, Balimau Putih, singly infected with RTBV or co-infected with RTBV + Rice tungro spherical virus (RTSV). These changes in virus accumulation resulted in differences in RTBV levels and incidence of infection. The virus levels were expressed relative to those of the susceptible variety and the incidence of infection was assessed at different weeks after inoculation. At a particular time point, RTBV levels in TN1 or Balimau Putih singly infected with RTBV were not significantly different from the virus level in plants co-infected with RTBV + RTSV. The relative RTBV levels in Balimau Putih either singly infected with RTBV or co-infected with RTBV + RTSV were significantly lower than those in TN1. The incidence of RTBV infection varied at different times in Balimau Putih but not in TN1, and to determine the actual infection, the number of plants that became infected at least once anytime during the 4 wk observation period was considered. Considering the changes in RTBV accumulation, new parameters for analyzing RTBV resistance were established. Based on these parameters, Balimau Putih was characterized having resistance to virus accumulation although the actual incidence of infection was >75%. PMID- 12898327 TI - Construction and characterization of a recombinant ovine lentivirus carrying the optimized green fluorescent protein gene at the dUTPase locus. AB - AdUTPase gene ( du) deleted ovine lentivirus (OvLV(Deltadu)) mutant, derived from Visna/maedi virus (VMV) molecular clone KV1772, was constructed. Subsequently, a copy of the optimized green fluorescent protein ( egfp) coding region was fused into the viral pol open reading frame (ORF) at the deleted du locus to generate viral mutant, OvLV(Deltadu-egfp). OvLV(Deltadu) reverse transcriptase (RT) activity and titer of infectious virus in goat synovial membrane (GSM) cell cultures were not affected compared to that of KV1772 and OvLV-85/34 strain (p < 0.05). By contrast, OvLV(Deltadu-egfp) RT activity and virus titer were lower than for KV1772 and OvLV(Deltadu) (p < 0.05). OvLV-85/34 RT in sheep monocyte derived macrophages (SMDM) was higher than that of KV1772, OvLV(Deltadu) and OvLV(Deltadu-egfp) (p < 0.05). The ability to prevent dUTP mis-incorporation into newly synthesized DNA was disrupted in OvLV(Deltadu) and OvLV(Deltadu-egfp) (p < 0.05). Immunoprecipitation demonstrated that GFP is expressed by OvLV(Deltadu egfp) at a low level. OvLV(Deltadu-egfp) retained egfp after 10 passages in cell culture.OvLV(Deltadu-egfp) was re-isolated in GSM cells from peripheral blood mononuclear (PBMN) cells of three of four OvLV(Deltadu-egfp)-inoculated lambs, but by contrast to the in vitro experiments OvLV(Deltadu-egfp) lost the insert. Priming with OvLV(Deltadu-egfp) did not prevent infection with pathogenic OvLV, but cell-associated viremia in a mock-infected contact control lamb was higher than in OvLV(Deltadu-egfp)-primed lambs. OvLV serum antibody titers increased steadily in OvLV(Deltadu-egfp)-inoculated lambs, but in a lamb from which OvLV(Deltadu-egfp) was not reisolated the antibody titer surpassed the negative/positive cut-off value only after challenge with OvLV-85/34. Because OvLV(Deltadu-egfp) is attenuated for pathogenicity in vitro, replicates in vivo and stimulates an antibody response, subsequent experiments need to address the likelihood of using OvLV(Deltadu-egfp) as an attenuated, live-virus vaccine to protect sheep against OvLV-induced disease when challenged with pathogenic OvLV. PMID- 12898328 TI - Virulence of infectious hematopoietic necrosis virus and Infectious pancreatic necrosis virus coinfection in rainbow trout ( Oncorhynchus mykiss) and nucleotide sequence analysis of the IHNV glycoprotein gene. AB - The outcomes of a coinfection of rainbow trout ( Oncorhynchus mykiss) with Infectious hematopoietic necrosis virus (IHNV) strain S46 and Infectious pancreatic necrosis virus (IPNV) strain S46 was determined after waterborne infection. Trout infected with the IHNV/IPNV.S46 sample, (a mixed sample containing equal infectious titers of the viruses) showed 50% less mortality than fish infected with either of the reference viruses alone. Forty-five days after the coinfection, IPNV antigens were detected by flow cytometry in 49 to 63% of the leukocytes from the surviving trout; whereas, only 9-15.6% of the leukocytes expressed IHNV viral antigens. IPNV was easily detected by reverse transcription polymerase chain reaction (RT-PCR), whereas, for IHNV, a second step of amplification of a 753 bp fragment corresponding to the internal sequences of the IHNV G gene was necessary to optimize viral detection. The sequence of the IHNV gene involved in virulence, the glycoprotein (G) gene, was determined for the IHNV.S46 and compared with other sequences available in the GenBank. Changes found were not located in the antigenic domains of the glycoprotein and were considered not significant. PMID- 12898329 TI - The complete nucleotide sequence of two distinct geminiviruses infecting cucurbits in Vietnam. AB - We have characterised two distinct geminiviruses that infect cucurbit cultivars in Vietnam. The genomes of both viruses consisted of two circular ssDNA components (DNA-A and DNA-B), with a genome arrangement and coding sequence typical of viruses in the Begomovirus genus in the family Geminiviridae. The sequence of DNA-A of one of the viruses was approximately 97% similar to Squash leaf curl virus-China (SLCV-Ch), for which a DNA-B has yet to be identified. We have named this virus Squash leaf curl virus-Vietnam (SLCV-Vn). The intergenic region of the SLCV-Vn DNA-B contained a 40 nt deletion between the putative AC1 TATA box and the stem loop. A second virus isolated from loofa in southern Vietnam was only 80% similar to SLCV-Vn over the complete DNA-A sequence, however the nucleotide sequence in the coat protein coding regions was 95% similar. We have named this virus Loofa yellow mosaic virus-Vietnam (LYMV-Vn). Other regions of the SLCV-Vn and LYMV-Vn genomes differed markedly, suggesting the coat protein coding region was recombinant. The DNA-B of both viruses were only 60% similar over the complete nucleotide sequence, although the encoded amino acid sequence of the BC1 gene was 90% identical. PMID- 12898331 TI - Experimental vertical transmission of Borna disease virus in the mouse. AB - We demonstrated the experimental vertical transmission of Borna disease virus (BDV) in pregnant BALB/c mice. Giessen strain He/80 of BDV was used in the present study. Six six-week-old mice were inoculated intraperitoneally with 10(5) 50% tissue culture infective doses (TCID50), and were bred immediately. Four pregnant mice were sacrificed under anaesthesia on the 10th and 14th days after vaginal plug formation. Nine newborns from two maternal mice were sacrificed under anaesthesia on the 7th day after birth. Positive signals with RT-nested PCR techniques for BDV p24-RNAs were seen in the fetuses, placentas and brains of all newborn mice. No immunopositivities for BDV p40 were found in the fetuses or placentas at 10 days' gestation. BDV p40 immunopositivities were found in neurons of the fetal brains and in decidual cells of the placentas at 14 days' gestation. They were also found in neurons of the brains of newborn mice. At 10 days' gestation, no positive signals for BDV p40 sense or antisense riboprobes were seen in the fetal brains or placentas. Positive signals were found in neurons of the fetal brains and decidual cells of the placentas at 14 days' gestation. Positive signals for BDV p40 sense and antisense riboprobes were found in almost all neurons throughout the brains of nine newborn mice. These results suggest that persistent infection with BDV in newborn mice may be induced by vertical transmission during gestation. PMID- 12898330 TI - Serological analysis of hemorrhagic fever with renal syndrome (HFRS) patients in Far Eastern Russia and identification of the causative hantavirus genotype. AB - Hemorrhagic fever with renal syndrome (HFRS) is endemic in East Asia and Europe. The disease is caused by several viruses belonging to the genus Hantavirus, including the Hantaan virus (HTNV), Seoul virus (SEOV), Dobrava Belgrade virus (DOBV), and Puumala virus (PUUV). Recently, HTNV-related viruses, Amur (AMR) and Far East (FE) genotypes were identified as causative agents of HFRS in Far Eastern Russia. To investigate the epidemiology of HFRS and virus transmission, we collected sera from 17 acute and 32 convalescent patients who were clinically diagnosed with HFRS in the Khabarovsk region of Far Eastern Russia, and detected anti-hantavirus antibodies using an ELISA that can differentiate the infected virus serotype using truncated hantavirus nucleocapsid protein antigen. Sixteen of the 17 acute phase patients had antibodies to hantavirus, and all the positive sera had higher optical densities for HTNV-specific antigen than for SEOV-, DOBV , or PUUV-specific antigens. The partial M segment of the viral genome was amplified from blood clots from three acute patients by PCR. The nucleotide sequences had closer identities to the FE genotype (>96%) than to the prototype HTNV (88 to 89%) or AMR genotype (81 to 83%). A phylogenetic analysis found that the virus sequences from the patients clustered with the FE type, and were distinct from the AMR type. Thirty-one of 32 convalescent patient sera had antibodies to HTNV-specific antigen. These data suggest that our ELISA system can detect HTNV-specific antibodies to the FE type, which may be responsible for most of the HFRS in Khabarovsk. PMID- 12898332 TI - Screening for T cell-eliciting proteins of Japanese encephalitis virus in a healthy JE-endemic human cohort using recombinant baculovirus-infected insect cell preparations. AB - The analysis of cell-mediated immune responses in virus-exposed but healthy individuals may contribute to define the features of the T cell response associated with resistance. We report, for the first time, on adaptive T cell responses to 5 largest of the 10 proteins that together constitute 76% of the coding potential of the Japanese encephalitis virus (JEV) genome in a naturally exposed healthy JE-immune human cohort. Fixed and sonified whole cell preparations of insect cells individually expressing recombinant prM, E, NS1, NS3 and NS5 proteins of JEV were used in vitro to stimulate lymphocytes from individuals who had experienced subclinical JEV infections. NS3-specific memory T cells were detected in up to 86% of the JEV-infected cohort whereas prM, E and NS1 each elicited reactions in approximately 45% among individuals tested, suggesting that NS3 is an important target for JEV-specific cell-mediated immune responses. Responses to NS5, the largest viral protein were in contrast the poorest, seen in only 13% of the cohort. Moreover, NS3 stimulated interferon gamma production in both CD4(+) and CD8(+) T cells indicating that a Th1 immune response to the NS3 protein may be a critical determinant of immune control of JEV infection. PMID- 12898334 TI - Molecular characterization and phylogenetic relationship of HERV-W family in the Macaca fuscata. AB - A human endogenous retrovirus, HERV-W, has recently been identified on human chromosome 7 and contains a single complete open reading frame putatively encoding an envelope protein. Its env gene was expressed in various tissues, and mainly in human placenta. We investigated env gene of the HERV-W family in the Macaca fuscata (Japanese monkey). The env gene expression was detected in various tissues (testis, prostate, kidney, cerebellum, thymus, placenta, intestine, stomach, ovary) of the Japanese monkey by RT-PCR. Southern blot analysis indicated that the monkey genome contained at least 15 copies of the HERV-W family. Using the PCR approach with the monkey genome, thirteen env fragments of the HERV-W family were identified and analyzed. These env fragments from monkey showed a high degree of sequence similarity (91-94%) to that of human HERV-W (AF072506). Putative amino acid sequences of the env fragments indicated multiple frameshifts and termination codons by deletion/insertion or point mutation in all clones identified in this study. Phylogenetic analysis of the env fragments derived from the monkey genome with those of the human genome showed a random sister relationship. PMID- 12898333 TI - Characterization of changes in the short unique segment of pseudorabies virus BUK TK900 (Suivac A) vaccine strain. AB - Mutant strains of pseudorabies virus (PRV) of reduced virulence, such as Bartha or BUK-TK900, have been used for vaccination purposes for many years. In contrast to the Bartha strain, BUK-TK900 has not been well characterised at the molecular level. The detailed analysis of this vaccine strain was urged by the fact of the isolation in Poland of field strains which were suspected to originate from BUK TK900. We characterised changes in the U(S) region of this strain, focusing our attention on gE and gI genes. The only deletion, about 300 bp, found in BamHI 7 fragment (covering most of the U(S) region) was located in the 28 K (US2) gene. BUK-TK 900 produced small plaques on all cell lines tested in our laboratory (SK6, Vero, MDBK, 3T3). The plaque size was restored to about 70% of wild type virus plaque size when growing BUK-TK900 virus on 3T3 complementing cell line expressing PRV gE and up to 100% when cell line producing gE and gI was used. Both gE and gI genes from BUK-TK900 and from some derivative field isolates have been amplified by PCR reaction but no deletions in these genes have been found. Molecular weight of gene products differed from wild type proteins: gE was bigger than wild type gE while gI was smaller. Both proteins were correctly recognised by all tested polyclonal and monoclonal antibodies. Radioimmunoprecipitation study showed that BUK-TK900 gE and gI interact forming a complex. The whole ORF of BUK-TK900 gE was sequenced and only few point mutations were found; only two of them led to changes of amino acids in the polypeptide chain. These were: methionine at position 124 replaced by threonine and glutamine at position 162 replaced by arginine. The introduction of first of these mutations (Met to Thr) to PRV wild type strain NIA-3 resulted in 22% reduction of plaque size. This result confirms the importance of this domain of gE for its function; it was found previously by others that deletion of amino acids 125 and 126 reduced virulence and neurotropism of PRV. More changes were found in BUK-TK900 gI sequence. Over 80% of these changes were located in the terminal 1/3rd of the sequence. Some of these mutations may have significant effect on the secondary structure of gI glycoprotein. The change of the secondary structure may be responsible for the decrease of gI stability and the observed reduction of gI molecular mass. PMID- 12898335 TI - Sequence analysis of the fragment of the phosphoprotein gene of Polish distemper virus isolates. AB - The nucleotide sequence analysis of the 429 bp fragment of the P gene of 11 Polish field isolates of Canine distemper virus (CDV), reference strains and other virus isolates available in the GenBank was the aim of the studies. High homology between all dog strains from east-southern region of Poland and reference strains of CDV was demonstrated. It was estimated as 97-100 % for CDV OND; 96.7-99.8 % for CDV-Rock; 96.7-99.8 % for CDV-LED and 96.3-97.9 % for A75 17. The 100 % homology of the nucleotide sequence was observed between CDV Pulawy 92, CDV Pulawy 97 and the reference CDV-OND. The homology between CDV-OND and viruses isolated from the mink and ferret was estimated as 97.7 % and 98.4 %, respectively. Virus strains isolated from blue foxes demonstrated the highest homology to CDV-OND - equal to 97.7 % for DV 79 and 99.5 % for DV 92. The fox isolate from 1992 had higher level of homology to dog isolates (96.5-99.5 %) than the strain isolated from the fox in 1979 (97.2-98.8 %). The phylogenetic tree has two main lineages representing two separated genetic groups: I containing PDV and II containing all distemper virus strains isolated from terrestrial carnivores. CDV strains isolated from dogs from Pulawy region between 1992-1998 and from the fox (DV 92) formed the separate lineage containing also reference strains. They differed from the native isolates from the mink and ferret as well as from Japanese strains of CDV. PMID- 12898336 TI - Epstein-Barr virus nuclear antigen-1 colocalizes with lamin B1 in the nucleoplasm and along the nuclear rim. AB - Epstein-Barr virus nuclear antigen 1 (EBNA-1) is essential for the maintenance of latent EBV plasmids, and is also a transcriptional regulator. Nuclear lamins, components of the nuclear lamina, have also been found in the nucleoplasm. We report here that EBNA-1 coincided with lamin B1 in the nucleoplasm and around the nuclear rim during S-phase by confocal microscopy of cells transfected with EBNA 1 in the absence of EBV plasmids. Lamin B1, which is rarely detected in nuclear soluble fractions, was detected in chromatin and nuclear matrix fractions of the EBNA-1-expressing cells. These observations suggest that EBNA-1 colocalizes with lamin B1 in the subnuclear sites. PMID- 12898338 TI - Viral nomenclature, where next? PMID- 12898339 TI - In memoriam: Harold S. Ginsberg (1917-2003). PMID- 12898340 TI - Protective effect of ginsenoside Rg1 on MPP+-induced apoptosis in SHSY5Y cells. AB - The neuroprotective mechanism of Rg1 was studied in this paper by means of its obvious anti-apoptotic effect on human SHSY5Y cells. SHSY5Y cells were treated with MPP+ (1-methyl-4-phenyl-pyridinium) for 72 hours to induce apoptosis. During the apoptosis, production of reactive oxygen species (ROS), activation of c-Jun N terminal kinase (JNK) and activation of caspase-3 were observed. The results showed that the signal transduction pathway of MPP+-induced apoptosis might be ROS to JNK, then to caspase-3. MPP+-induced apoptosis in SHSY5Y cells was obviously inhibited in both NAC (N-acetylcysteine) pretreated groups and Rg1 pretreated groups. Meanwhile, compared to that of the controls, our results showed decreased level of ROS, less JNK activity and lower expression of cleaved caspase-3 in pretreated NAC groups and in Rg1 pretreated groups. The protection by Rg1 might be mediated by removing of ROS. The removal of ROS might inhibit the activity of JNK and the expression of cleaved caspase-3. These results suggest that ginsenoside Rg1 may take effect through its anti-apoptotic activity in neurodegenerative diseases. PMID- 12898341 TI - Long-lasting astrocyte reaction to persistent Junin virus infection of rat cortical neurons. AB - Immunoperoxidase labeling was performed in histological sections from rat brain harvested during acute (10-30 days), clinically inapparent (90-270 days) and late (450-540 days) stages of Junin virus-induced neurological disease. In frontoparietal cortex, count of viral antigen (+) neurons peaked during the acute period (27.7+/-6.8), dropped within the intermediate (4.8+/-4.0 to 1.4+/-1.1) and increased (7.6+/-4.3) at the onset of the late neurological syndrome. In infected vs. control rats, the number of GFAP (+) astrocytes maximized during the acute stage (19+/-4 vs. 11+/-5), and from the end of the intermediate (27+/-5 vs. 21+/ 5) up to the late (37+/-7 vs. 26+/-6) periods. In turn, surface density of GFAP (+) material in infected samples peaked at 0.196+/-0.066, while it failed to exceed 0.090+/-0.043 in controls. Both astrocyte hypertrophy relapsing into chronicity, as depicted by surface density, and astrocyte hyperplasia preceding the onset of the late neurological syndrome, support their pathogenic contribution to disease expression. PMID- 12898342 TI - Effects of monoamine oxidase inhibitors on the diethyldithiocarbamate-induced enhancement of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine toxicity in C57BL/6 mice. AB - Diethyldithiocarbamate (DDC) is known to potentiate the neurotoxicity of 1-methyl 4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). The aims of the present study were to provide biochemical, pathological and behavioral evidence for the degeneration of dopamine (DA) neurons in C57BL/6 strain mice treated simultaneously with DDC and MPTP, and to evaluate the effects of monoamine oxidase (MAO) inhibitors on DDC enhanced MPTP toxicity. DDC (400 mg/kg)+ MPTP (30 mg/kg) treatment decreased significantly the levels of striatal DA and its metabolites and induced bradykinesia. In mice treated with DDC+MPTP, degenerative areas were found in striatum, substantia nigra and tuberculum olfactorium by assessment of the binding of [125I]RTI-121, a DA transporter ligand. Pretreatment with a MAO-B inhibitor selegiline prior to the administration of DDC and MPTP completely inhibited the decrease in the levels of DA and its metabolites, bradykinesia and degeneration of dopaminergic nerve terminals. In contrast, the protective action of clorgyline was not clearly observed in this model system. PMID- 12898343 TI - Recovery of motor function and dopaminergic parameters in a mouse model of Parkinson's disease induced by co-administration of 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine and diethyldithiocarbamate. AB - Diethyldithiocarbamate (DDC) enhances the neurotoxicity of 1-methyl-4-phenyl 1,2,3,6-tetrahydropyridine (MPTP). We studied the time course of dopaminergic parameters and motor function of MPTP+DDC-lesioned C57BL/6 mice, a model of Parkinson's disease. MPTP+DDC-lesioned mice showed a decrease in dopamine (DA) and its metabolites contents in their striata 1, 3 and 6 weeks after MPTP+DDC treatment, compared with those of each control group. The partial and significant recoveries in DA, 3,4-dihydroxyphenylacetic acid, and homovanillic acid contents were also observed after 6 weeks, compared with those at 1 week after treatment. In addition, bradykinesia due to DA depletion was observed in mice 1 week after MPTP+DDC-treatment, but it was not significant 3 weeks after the treatment. l DOPA alone and a co-administration of l-DOPA and a monoamine oxidase-B inhibitor selegiline improved bradykinesia of this model, also suggesting that bradykinesia observed in the model was mediated to dopaminergic deficiency. On the other hand, the serotonin content increased slightly but significantly after 3 or 6 weeks, suggesting compensatory activation of the serotonergic system against DA depletion. Thus, the partial recovery of dopaminergic parameters, the recovery of motor function and the compensatory activation of the serotonergic system were observed in this model 3-6 weeks after MPTP+DDC treatment. PMID- 12898344 TI - Influence of age, gender, education and dexterity on upper limb motor performance in Parkinsonian patients and healthy controls. AB - Finger tapping, the most widely used test for evaluating motor dysfunction in Parkinson's disease (PD), was found to react sensitively to disease specific factors like disease severity and changes in medication. A possible interference caused by disease unrelated demographic factors--age, gender, education and dexterity--however has not yet been studied systematically. Various components of tapping performance of 187 healthy subjects and 200 PD patients were assessed by means of the BRAIN TEST, a digitalized test battery. The effects of demographic factors--above all education and age--were found to be significant. These influences generally affect different aspects of movement to a different extent, with speed and akinesia being affected more severely than dysmetria and arrhythmokinesis. Our study suggests that whenever precise assement of upper limb motor performance is needed, specific corrections for these demographic factors in both healthy controls and PD patients are necessary. PMID- 12898345 TI - L-DOPA biotransformation: correlations of dosage, erythrocyte catechol O methyltransferase and platelet SULT1A3 activities with metabolic pathways in Parkinsonian patients. AB - The objectives of this study were to determine (1) the effects of dose and drug absorption on pathways of biotransformation of L-DOPA in Parkinsonian patients treated with Sinemet, and (2) the extent to which genetically-determined variations in the activities of erythrocyte catechol O-methyltransferase and/or platelet phenol sulfotransferase might be reflected in individual differences in L-DOPA metabolism. In the 19 patients studied, there were negative correlations between dosage or absorption and extent of O-methylation and of sulfation of L DOPA or its metabolites. Levels of activity for erythrocyte COMT were also reflected in individual variation in the metabolism of L-DOPA. In contrast, differences in platelet phenol sulfotransferase were not reflected in differences in sulfation of L-DOPA or of its metabolites. If such a relationship did exist, it might have been obscured by the effects of high dosage of L-DOPA, effects which might have resulted from a deficiency of the sulfation cosubstrate 3' phosphoadenosine 5'-phosphosulfate in patients taking higher doses of drug. PMID- 12898346 TI - Modifications of plasma and platelet levels of L-DOPA and its direct metabolites during treatment with tolcapone or entacapone in patients with Parkinson's disease. AB - We compared--retrospectively--the effects of a 3-month therapy with catechol- O methyltransferase (COMT) inhibitors tolcapone (100 mg, t.i.d.) and entacapone (200 mg, t.i.d.), on L-DOPA metabolism in two groups of parkinsonian patients with motor fluctuations. Plasma and platelets concentrations of L-DOPA and its direct metabolites, dopamine and 3- O-methyldopa (3-OMD), were measured before starting treatment, after two weeks and at the end of treatment. Patients treated with tolcapone showed significant increases in plasma and platelet L-DOPA levels and marked reduction of plasma and platelet 3-OMD levels, both at short- and long term. Entacapone did not modify L-DOPA levels, while inducing a less marked reduction of plasma and platelet 3-OMD concentrations, with respect to tolcapone, at both time points. Both drugs were similarly effective in increasing plasma and platelet levels of dopamine. These results confirm the different profiles of activity of the two drugs, with tolcapone proving more effective on both the intra- and extra-cellular levels of L-DOPA and 3-OMD. PMID- 12898347 TI - Regional serotonin transporter availability and depression are correlated in Wilson's disease. AB - In patients with Wilson's disease (WD), depression is a frequent psychiatric symptom. In vivo neuroimaging studies suggest that depression and other neuropsychiatric disorders are associated with central serotonergic deficits. However, in vivo measurements of serotonergic neurotransmission have not until now been performed in patients with this copper deposition disorder. The present prospective study revealed that depressive symptomatology is related to an alteration of presynaptic serotonin transporters (SERT) availability as measured by [123I]-2beta-carbomethoxy-3beta-(iodophenyl)tropane ([123I]beta-CIT) and high resolution single-photon emission computed tomography (SPECT). SERT imaging with [123I]beta-CIT-SPECT could therefore become a useful tool for diagnosis and therapy monitoring in depressed WD patients. PMID- 12898348 TI - Characterization of the effect of ganstigmine (CHF2819) on amyloid precursor protein metabolism in SH-SY5Y neuroblastoma cells. AB - We have investigated the effect of ganstigmine (CHF2819), a novel geneserine derived acetylcholinesterase (AChE) inhibitor, on the expression and metabolism of the amyloid precursor protein (APP) in neuroblastoma cell line SH-SY5Y. The rationale was based on the suggestion that cholinergic activity may also be involved in the regulation of APP metabolism. We studied the acute effect on APP metabolism following the secretion of sAPPalpha in the conditioned medium of cells. Following short term treatment (2h), ganstigmine promoted a slight increase in the release of sAPPalpha, the maximal effect approaching on average 1.5 fold baseline value. The data obtained in the long term experiments demonstrate that continuous inhibition of AchE obtained with 100 nM ganstigmine following an exposure of 24 hours did not influence APP isoforms expression. However, the compound appeared to increase the constitutive release of sAPPalpha, with a mechanism that is derived from an indirect cholinergic stimulation. PMID- 12898349 TI - Decreased lysophosphatidylcholine/phosphatidylcholine ratio in cerebrospinal fluid in Alzheimer's disease. AB - Choline containing phospholipids are essential for the integrity of the'cell'membrane. Minor changes in the lysophosphatidylcholine (lyso PC)/phosphatidylcholine (PC) ratio may lead to neuronal damage and cell loss. Several studies have shown protein and lipid oxidation in Alzheimer's disease (AD) affected brain regions. Amyloid-beta peptides may induce free-radical oxidative stress which normally is counteracted by anti-oxidant defense mechanisms. We hypothesize that oxidation may lead to changed concentrations of choline containing phospholipids in cerebrospinal fluid (CSF) of AD patients, because of the susceptibility of the unsaturated acyl-chains of PC for oxidation. PC and lyso-PC were determined in CSF of AD patients (n=19) and subjects with subjective memory complaints without dementia (n=19) by tandem mass spectrometry. No differences in total PC concentrations were observed between both study groups. Furthermore, we could not demonstrate different concentrations of PC species containing linoleic acid and PC species containing arachidonic acid. Interestingly, lyso-PC concentrations tended to be lower while the lyso-PC/PC ratio was significantly decreased in CSF of AD patients compared to controls (0.36% versus 0.54%; P=0.017). A comparable decrease was found for the lyso-PC/PC ratio for PC containing linoleic acid (P=0.022) or arachidonic acid (P=0.010), respectively. The lower lyso-PC/PC ratio in CSF of patients with AD may reflect alterations in the metabolism of choline-containing phospholipids in the brain in AD, and suggests that PC species containing linoleic acid or arachidonic acid are equally involved. PMID- 12898350 TI - Minimally invasive ventral spondylodesis for thoracolumbar fracture treatment: surgical technique and first clinical outcome. AB - A new instrumentation system for ventral stabilization of the spine that can be used for an endoscopic and minimally invasive approach was developed. We describe the implantation technique and report on the first clinical results. This prospective study covers the first 45 patients to undergo this new technique since it was introduced in 1999. In all patients the operation was successfully performed in a completely minimally invasive procedure. Mono- and bisegmental stabilization was performed mainly at the thoracolumbar junction after initial posterior instrumentation in most cases. Lesions varied from fresh/old fractures to metastases (T5-L3). Pre- and postoperative follow-up included clinical examination and radiological visualization via X-ray and computed tomographic scan. Our experience with this minimally invasive procedure demonstrated the feasibility of the method. PMID- 12898351 TI - Revisiting the rDNA sequence diversity of a natural population of the arbuscular mycorrhizal fungus Acaulospora colossica. AB - In 1999, the diversity of a field population of the arbuscular mycorrhizal (AM) fungus Acaulospora colossica was characterized using DNA sequence data. Since 1999, AM fungal sequences have accumulated rapidly within public databases. Moreover, novel phylogenetic tools have been developed and can be used to interpret the data. A second analysis of those sequences collected in 1999 demonstrates that while the majority of the sequences are, in fact, sequences of A. colossica; a minority of the sequences still cannot be identified with confidence. Those sequences identified as A. colossica can be used to show that (1) the nuclear rDNA ITS regions are remarkably diverse, and (2) sequences isolated from different spores of the same site may be more closely related to each other than to sequences of other sites, so that the genetic diversity of an AM fungal field population may be spatially structured; however, identical sequences can also be recovered from different sites. PMID- 12898353 TI - Emerging concepts for the mechanism of alcoholic pancreatitis from experimental models. AB - The pathophysiologic mechanisms that underlie acute and chronic pancreatitis arising from alcohol abuse are poorly understood. The reasons for this state of knowledge result historically from a lack of models for experimental investigation. Ethanol feeding alone, even at high doses, has minimal and inconsistent effects on morphologic findings in the pancreas in experimental animals. This experience, plus the fact that alcohol abuse causes pancreatic pathology in only a minority of patients, suggest that ethanol acts to sensitize the pancreas to the deleterious effects of other stimuli. In this article, we discuss findings to support this concept of ethanol as a sensitizing agent and experimental models developed that can be used to investigate the effects of ethanol on the pathologic processes of pancreatitis. These pathologic processes include inflammation, cell death, intrapancreatic digestive enzyme activation, and fibrosis. PMID- 12898354 TI - Comparison of gastric peristalsis inhibition by scopolamine butylbromide and glucagon: evaluation by electrogastrography and analysis of heart rate variability. AB - BACKGROUND: Activation of glucagon receptors of the smooth muscle membrane suppresses gastric peristalsis. We evaluated autonomic nervous activity by two methods, electrogastrography (EGG) and analysis of heart rate variability, to compare the inhibiting effects of glucagon and scopolamine butylbromide on gastric peristalsis. METHODS: Heart rate variability, EGG, and blood catecholamine levels were measured before and after administration of glucagon (G group), scopolamine butylbromide (SB group), or physiological saline (C group). Autonomic nervous function was evaluated using spectral analysis of heart rate variability, and low frequency (LF) and high frequency (HF) power; the LF/HF ratios were also determined. RESULTS: After administration of scopolamine butylbromide, HF power, an index of parasympathetic nervous activity, decreased; and the LF/HF ratio, an index of sympathetic nervous activity, increased. In contrast, no significant change was observed in autonomic nervous activity after administration of glucagon. The peak power amplitudes of the EGG decreased significantly in the G and SB groups after intramuscular injection, but the difference between the groups was not significant. Furthermore, the dominant frequency increased significantly in the G and SB groups after injection. Serum catecholamine levels showed no significant changes after administration of scopolamine butylbromide or glucagon. CONCLUSIONS: Inhibition of gastric peristalsis by glucagon via glucagon receptors on smooth muscles did not influence autonomic nervous activity, unlike the results obtained after administration of scopolamine butylbromide. Therefore, glucagon may be safe for use with elderly patients and those with cardiopulmonary complications. PMID- 12898355 TI - Differential expression of EGF receptor in the pig duodenum during the transition phase from maternal milk to solid food. AB - BACKGROUND: The aim of this investigation was to study the cell type-specific expression of epidermal growth factor receptor (EGF-R) and to evaluate changes of the EGF-R distribution during transition from maternal milk to solid food in the gastrointestinal tract of young piglets. METHODS: Duodenal tissue probes from six pigs were taken 2 days before (-2d) and 2 days (+2d) and 14 days (+14d) after transition from milk to solid food. The specimens were fixed in methanol/glacial acetic acid (2 : 1). A monoclonal antibody against EGF-R was used to examine the pattern and topographical shift of EGF-R. To assess a possible correlation between EGR-R-positive cells and mitotic activity, the mitotic index (MI) were evaluated based on expression of the Ki-67 antigen. RESULTS: A significant change in the topographical and cellular distribution of the EGF-R could be successfully determined during the transition period. The highest immunoreactivity for EGF-R was found in enterocytes 2 days before transition from maternal milk, predominantly around the villous tips. Two days after transition consistent staining along the villi and crypts could be demonstrated. Fourteen days later the expression was significant lower around the villous tips and was more concentrated in Brunner's glands. Additionally, distinct expression of the receptor is selectively found in stimulated goblet cells. The analysis of the mitotic activity during the transition period shows that cells that highly express the EGF-R have a rather low proliferation rate. CONCLUSIONS: Our findings suggest that EGF plays an important role in cell differentiation (rather than cell proliferation) in young animals, and it may be involved in stimulating mucus secretion. PMID- 12898356 TI - Elucidation of cytomegalovirus disease recurrence in an HIV-1-positive patient. AB - BACKGROUND: Cytomegalovirus (CMV) is responsible for the most common opportunistic viral infection in patients with acquired immunodeficiency syndrome (AIDS). The colon is a common site for CMV infection in patients positive for the human immunodeficiency virus (HIV). The clinical diagnosis of CMV infection is based on the characteristic endoscopic appearance of extensive ulceration of the gastric mucosa. METHODS: A 54-year-old HIV-positive man visited our hospital because of high fever. The patient presented with diarrhea, and colonoscopy was performed. Ganciclovir was administered, with a good clinical response. However, the patient complained of the same symptoms again 6 months later. Nested polymerase chain reaction (PCR) was performed on all the patient's samples to detect CMV, followed by sequencing of the UL97 gene for CMV resistance detection. RESULTS: The PCR tests for Legionella, Chlamydia pneumoniae, Pneumocystis carinii, and Aspergillus were negative. DNA preparations from biopsy specimens of areas of colon ulceration were positive for CMV. Six months after treating the colon ulcerations, the PCR for CMV was positive, and the possible emergence of CMV mutants conferring ganciclovir resistance was examined. Direct sequencing of the PCR products revealed the known V594 mutation in the UL97 gene predisposing for ganciclovir resistance as well as the polymorphisms (579, GGC-->GGT) and (598, GGT-->GGC) in all samples tested. CONCLUSIONS: In summary, molecular biology methods can be used for early detection of CMV in characteristic colonic lesions in HIV-1-positive patients. Furthermore, detection of mutant strains resistant to antiviral drugs as well as polymorphisms elucidate the natural history of the infection. PMID- 12898357 TI - Clinical features and pattern of indeterminate colitis: Crohn's disease with ulcerative colitis-like clinical presentation. AB - BACKGROUND: Although an accurate diagnosis of inflammatory bowel disease (IBD) and differentiation between ulcerative colitis (UC) and Crohn's disease (CD) can be made in most patients, it is sometimes impossible to distinguish UC from CD even after thorough pathological study. Recently, clinicians have used the term indeterminate colitis (IC) for patients with features of both diseases that overlap temporarily or persistently. The frequency, reasons, and outcome of patients with a clinical diagnosis of IC based on radiological, endoscopic, and histopathological findings were investigated retrospectively. METHODS: Based on records of 735 patients with IBD, IC was defined as having features of both UC and CD, with differentiation from each other impossible at least once during the observation period (average 6.8 years) based on diagnostic criteria using endoscopic, radiological, and histological findings. RESULTS: Twenty-three patients were identified as having IC. They were classified into three patterns according to the clinical course and the final diagnosis: (1) UC changing to CD (n = 8); (2) CD changing to UC (n = 5); and (3) UC or CD (n = 10). The frequency of IC was 24.5%-43.4% of colitis-type CD (n = 53), 2.3%-6.5% of all CD (n = 352), and 3.1% of IBD (n = 735). The reasons for the indetermination were temporary (56.5%) or persistent (43.5%) overlapping of UC-like and CD-like presentations. Treatment of IC was inappropriate in only two patients, and the prognoses of all patients except one were fairly good. CONCLUSIONS: Overlapping of UC-like presentations (persistent bloody stool and diffuse colitis) was frequently observed with Crohn's colitis but less so in CD patients during their clinical course. The basis of differentiation and treatment of IC needs more attention. PMID- 12898358 TI - Clinical features of hepatitis B virus genotype A in Japanese patients. AB - BACKGROUND: Hepatitis B virus (HBV) genotype A is predominant in northern Europe and central Africa. In the present study, we examined the clinical features associated with HBV genotype A disease in the Tokyo metropolitan area. METHODS: We investigated 53 cases of HBV surface antigen (HBsAg)-positive Japanese patients with HBV genotype A. The 53 cases were further classified as to their serum alanine aminotransferase (ALT) status being within the normal range (asymptomatic carriers, n = 17), chronic hepatitis (n = 15), liver cirrhosis (n = 4), and acute hepatitis (n = 17). RESULTS: Chronic hepatitis patients had significantly higher HBV DNA levels (P = 0.003) and hepatitis B e antigen (HBeAg) positivity rates at the initial visit than did asymptomatic carriers or patients with liver cirrhosis (P = 0.003 and P = 0.054, respectively). The efficacy of treatment (HBeAg seroconversion rate) was 75% in 12 chronic hepatitis patients, which was excellent. A family history of HBsAg positivity was identified in eight (15%) families (five asymptomatic carriers, three with chronic hepatitis). However, none of the mothers in the study was positive for HBV genotype A. CONCLUSIONS: Maternal transmission of HBV has often been reported in Japan, but our present findings suggest that horizontal infection of HBV genotype A is more prevalent in the Tokyo metropolitan area. Our data indicate that HBV genotype A exhibits a mode of infection different from that of conventional HBV previously seen in Japan. PMID- 12898359 TI - Abnormalities of tumor suppressor gene p16 in pancreatic carcinoma: immunohistochemical and genetic findings compared with clinicopathological parameters. AB - BACKGROUND: Abnormalities of the tumor suppressor gene p16 have been reported in a variety of human tumors but are rare in pancreatic carcinoma except for cancer cell lines and xenografts. Their clinicopathological significance remains unknown. The purpose of this study was to examine immunohistochemical and genetic alterations of p16 in primary pancreatic carcinoma tissues and to investigate the relation between abnormalities of p16 and clinicopathological parameters to elucidate their clinicopathological significance. METHODS: We investigated p16 expression in 60 pancreatic carcinoma cases by immunohistochemistry using a monoclonal antibody clone G175-405. In addition, we analyzed genetic alterations of the p16 gene using DNA extracted from microdissected tissue of pancreatic carcinoma, by polymerase chain reaction, nonradioisotopic single-strand conformation polymorphism (non-RI-SSCP), DNA sequencing, and hypermethylation analyses using restriction enzymes. We compared the abnormalities of p16 alterations with clinicopathological parameters to elucidate their significance. RESULTS: On immunohistochemical study, staining for p16 protein was strongly positive in 22 (37%) of 60 pancreatic carcinoma cases, weakly positive in 24 (40%), and negative in 14 (23%). In contrast, p16 mutations were recognized in 9 (15%) of the 60 pancreatic carcinoma cases. The incidence of p16 mutations was 2 (9%) in 22 cases of pancreatic carcinoma with strongly positive staining, 4 (17%) in 24 with weakly positive staining, and 3 (21%) in 14 with negative staining. Hypermethylation of p16 was detected in the two pancreatic carcinoma cases with weakly positive staining, although homozygous deletions were not found in any case. There was no significant correlation between the expression of p16 protein and any of the clinicopathological parameters. However, there was a tendency for the tumor to be larger in patients with decreased expression of p16 protein than in those with normal expression levels. In contrast, the tumor was significantly larger and the survival period significantly shorter for patients with pancreatic carcinoma with p16 mutation or hypermethylation than for those with pancreatic carcinoma with an intact p16 gene (P < 0.05). CONCLUSIONS: These findings suggest that p16 alterations may participate in the aggressiveness of pancreatic carcinoma. PMID- 12898360 TI - Pancreatic regeneration after ethionine-induced acute pancreatitis in rats lacking pancreatic CCK-A receptor gene expression. AB - BACKGROUND: We examined the effects of cholecystokinin (CCK) on the development of ethionine-induced pancreatitis and pancreatic recovery. We used Otsuka Long Evans Tokushima Fatty (OLETF) rats, a model lacking pancreatic CCK-A receptor gene expression. METHODS: Ethionine-induced pancreatitis was induced in the 7 week-old male OLETF rats and in a control group that does not lack the pancreatic CCK-A receptor, Long-Evans Tokushima Otsuka (LETO) rats. The two groups were maintained on a low-protein diet for 11 days. During the last 4 days of the low protein diet, dl-ethionine 20 mg/100 g body weight was administered intraperitoneally once daily. Histologic and biochemical examinations of the pancreas were performed, and plasma CCK concentrations were measured on days 1, 4, and 7 after the last ethionine administration. RESULTS: Pancreatic histologic scores for inflammation, hemorrhage, and necrosis in the LETO and OLETF rats were highest on days 1 and 4, respectively. Pancreatic weight, DNA content, and protein level per DNA content in both groups decreased during the low-protein diet, and recovery signs were delayed in the OLETF rats. The highest plasma CCK concentrations in the LETO and OLETF rats were reached on days 1 and 4, respectively. CONCLUSIONS: Ethionine-induced pancreatitis developed in the OLETF rats, and their pancreatic regeneration was delayed in comparison to that in the LETO rats. Our results suggested that CCK plays an important role in the development of pancreatitis as well as in the pancreatic repair process. PMID- 12898361 TI - Hemorrhagic rectal ulcer caused by a deformity of the coccyx complicated by normal delivery. AB - A 25-year-old woman delivered her first-born child weighing 3400 g by full-term normal delivery. Bloody stool developed 3 days after delivery. Colonoscopic examination revealed two linear ulcers parallel to the anal column on the posterior side of the rectal ampulla about 5 cm from the anal ring. Coagulated blood was detected at the ulcer bottom, suggesting the site of hemorrhage. Bowel rest in the absence of oral intake resulted in the disappearance of bloody stool. Three days later colonoscopic examination revealed improvement in the ulcer. There was no sign of constipation. A lateral pelvic radiograph showed that the apex of the coccyx was flexed anteriorly. The parturient canal may have physically pressed the rectum against the apex of the coccyx during delivery, causing mucosal injury. Only a few studies have reported the complication of a rectal ulcer due to injury during delivery. Furthermore, none of the previous studies has reported a rectal ulcer caused by deformity of the coccyx. PMID- 12898362 TI - Poorly controlled ulcerative colitis treated by colectomy during remission induced by extracorporeal leukocyte removal therapy. AB - Both monocyte-granulocytapheresis (M-GCAP) and leukocytapheresis (LCAP) are categorized as extracorporeal leukocyte removal therapies (ECCTs). These therapies have been recognized as efficient adjuncts for patients of steroid resistant ulcerative colitis (UC). This study aimed to consider the adaptation and the limitation of these new therapies from the clinical standpoint based on a case of UC showing strong resistance to high-dose continuous steroid injection therapy. The patient successfully underwent a scheduled colectomy while maintaining remission after applying M-GCAP and LCAP independently. Surgical therapy was chosen because of a deep ulcer in the patient's sigmoid colon, which was assumed to constitute a future risk for perforation. This case suggests that combining ECCT with steroid therapy can maintain such poorly controlled and high risk UC patients safely for the scheduled colectomy while improving the prognosis by reducing the dosage of steroid efficiently prior to operation. PMID- 12898363 TI - Effect of a tyrosine kinase inhibitor STI571 in a patient with hepatic metastases from a duodenal gastrointestinal stromal tumor. AB - Gastrointestinal stromal tumors (GISTs) are the most common mesenchymal tumors. The molecular etiology is the result of mutations in the c-Kit gene. The mutant c Kit proteins, which are activated without a stem cell factor, contribute to the tumor development. STI571 selectively inhibits c-Kit, BCR-ABL, and PDGFR tyrosine kinases. Based on this potential to inhibit critical c-Kit function in GISTs, case studies have reported effective outcomes following treatment with STI571. This case report describes a highly effective use of STI571 in a 54-year-old woman with multiple liver metastases from a GIST originating in the duodenum. PMID- 12898364 TI - Scintigraphic study of regenerative nodules due to fulminant hepatic failure. AB - We report the case of a 25-year-old woman with fulminant hepatic failure (FHF). Liver scintigraphy using (99m)Tc-galactosyl human serum albumin (GSA) and (99m)Tc phytate produced interesting findings; regenerative nodules appeared as nodules of increased accumulation of (99m)Tc-GSA, whereas these nodules were expressed as defects of accumulation of (99m)Tc-phytate. These scintigraphic findings suggested that the functions of hepatocytes in regenerative nodules were maintained, whereas those of Kupffer cells were impaired. Although (99m)Tc-GSA scintigraphy indicated hepatic functional reserve enough to survive, she died despite intensive therapy including plasma exchange. Based on this case, it is recommended that not only (99m)Tc-GSA scintigraphy but also (99m)Tc-phytate scintigraphy is required to evaluate the prognosis of patients with FHF. PMID- 12898365 TI - Varicose bleeding after liver transplantation in a patient with severe portosystemic shunts. AB - Recipients for liver transplantation often have portosystemic shunts due to portal hypertension. It is an important problem whether such shunts should be ligated during operations. Ligating the shunts seems of benefit for increasing portal blood flow to the liver, but it is sometimes difficult technically, and it is invasive to the patient. We experienced a recipient with huge portosystemic shunts and no esophageal varices before living-related liver transplantation. Some shunts were ligated during operation to increase portal blood flow to the graft. Unfortunately, the patient suffered severe bleeding from esophagogastric varices after he underwent retransplantation owing to accidental liver failure. Based on our experience, extreme care should be exercised to avoid varicose bleeding after ligating the portosystemic shunts of liver transplantation patients. PMID- 12898366 TI - Early gallbladder carcinoma associated with primary sclerosing cholangitis and ulcerative colitis. AB - Patients troubled with primary sclerosing cholangitis (PSC) and ulcerative colitis (UC) are at high risk for cholangiocarcinoma, whereas cancer of the gallbladder (GBC) is rarely reported to develop in that population. A Japanese man aged 62 years with a 14-year history of PSC and UC had been found to have a protruding lesion of the gallbladder by screening sonography. The preoperative examination suggested the lesion to be GBC at an early stage. Pathology examination after cholecystectomy proved that the lesion was papillary adenocarcinoma localized in the mucosal layer. Although the prognosis of GBC is poor, the outcome of cholecystectomy against early GBC is relatively good. Early detection of the tumor is required for a better prognosis of patients with GBC. According to the review of the literature, PSC and UC patients are regarded as a high-risk group not only for cholangiocarcinoma but also GBC. It is advocated that clinicians perform repeated radiographic examinations including sonography for patients with PSC and UC even if the diseases are being controlled. PMID- 12898367 TI - Improvement of overexpression of endothelin B receptor in human cirrhotic liver by interferon therapy. PMID- 12898368 TI - Barriers to cancer pain management: home-health and hospice nurses and patients. AB - GOALS: Undertreatment of cancer pain remains a major health-care problem. We utilized focus groups of hospice and home-health nurses and patients to elucidate factors contributing to inadequate pain management and to generate solutions for closing the gap between the current reality and optimal pain management. PATIENTS AND METHODS: Focus groups were conducted among hospice and home-health-care nurses (two groups; n=22) and patients (six groups; n=54) using a standardized question guide. Audiotapes were transcribed and analyzed using NUD*IST software. Themes discovered among patients and nurses were analyzed for similarities and differences. MAIN RESULTS: Of 22 participants in the two home-health and hospice nurses focus groups, all were white women, the average age was 43 (range 29-64) years, and the average number of years in nursing was 21 (range 8-47) years. Of 54 participants in the six cancer patient focus groups, 80% were women, the average age was 54 (range 25-76) years, and 76% were white. Fifty-four percent of patients reported a history of pain associated with their cancer, and almost 30% had pain that they rated as 8 or higher on the pain scale. Barriers to adequate pain management fell into four categories: fears; attitudes, beliefs, and values; patient and provider behaviors; and structural barriers. Patients and nurses reported similar barriers to pain management; however, patients identified more barriers related to provider behavior and structure of the health care system. CONCLUSIONS: This study identified several barriers to cancer pain control not previously identified in the literature. Strategies to improve cancer pain control are suggested. PMID- 12898369 TI - Family caregiver knowledge of treatment intent in a longitudinal study of patients with advanced cancer. AB - GOALS OF WORK: Caregivers have become part of a triad of care and frequently attend patient consultations in the ambulatory cancer setting. Effective caregiving and decision making require that they understand the course of the disease and the changing treatment goals. This study sought to evaluate caregiver perception of treatment intent. PATIENTS AND METHODS: A cohort of 317 subjects (181 patients and 136 caregivers) from The Canberra Hospital's Cancer Services were followed for 6 months. Caregiver understanding of patient treatment intent was measured over time together with sources of information. MAIN RESULTS: Most caregivers understood that the illness was life-threatening (92% at week 12) and that treatment goals were to control illness and improve quality of life. Only half understood that treatment was noncurative (48% at week 12); 27% were unsure and 25% believed that treatment would cure. A high proportion of caregivers identified the specialist as the source of information (77%) and almost half also included the general practitioner (47%). These figures remained fairly constant over time. There were significant gender and age differences in understanding. At baseline, more women than men had an accurate perception of treatment intent and these numbers increased over time. Men's perception did not change. CONCLUSIONS: Caregivers' ability to fully engage in the task of caring for those with a terminal illness may be hampered by their lack of understanding of the treatment patients receive. PMID- 12898371 TI - Caring for dying patients--what are the attitudes of medical students? AB - Students entering medical school today will encounter an ageing population and a higher incidence of patients with terminal and life threatening illness. This study was carried out to determine the attitudes of pre-clinical medical students towards the care of patients for whom a cure is not possible. All students were invited to complete a 23-item questionnaire prior to any teaching in palliative care. Of the 186 students who participated in the study, 151 (79%) completed the pre-teaching questionnaire (males=59; females 90; median age 20 years; range 19 27]. Attitudes towards chronically ill and dying patients were generally positive. When comparing age with attitudes it was found that increasing age was associated with a more positive view of caring for patients with chronic or terminal illness, a more positive view of listening to patients reminisce and a more positive view of patients dying at home. Caring for patients at the end of life can be one of the most rewarding aspects of being a doctor. This study suggests that the majority of medical students have a positive attitude towards patients with chronic and terminal illness-the trend for encouraging older students to enter medicine may be an influencing factor. PMID- 12898370 TI - Feasibility, physical capacity, and health benefits of a multidimensional exercise program for cancer patients undergoing chemotherapy. AB - Cancer patients frequently experience considerable loss of physical capacity and general wellbeing when diagnosed and treated for their disease. The aim of this study was to evaluate the feasibility, physical capacity, and health benefits of a multidimensional exercise program for cancer patients during advanced stages of disease who are undergoing adjuvant or high-dose chemotherapy. The supervised program included high- and low-intensity activities (physical exercise, relaxation, massage, and body-awareness training). A total of 23 patients between 18 and 65 years of age (median 40 years) participated in groups of seven to nine patients for 9 h weekly for 6 weeks. Physical capacity in terms of repetition maximum (RM) and maximal oxygen uptake (VO(2)max), physical activity level and psychosocial wellbeing (EORTC QLQ-C30, SF-36, HAD) were compared prior to and after completion of the program. The program was safe and well tolerated. The completion rate was 85.2%. Highly significant increases in physical capacity (1RM, VO(2)max) and an improved level of physical activity were achieved. Quality of life and general wellbeing assessments indicated improvements in several measures, but without reaching significance. It is concluded that an exercise program, which combines high- and low-intensity physical activities, may be used to prevent and/or minimize physical inactivity, fatigue, muscle wasting and energy loss in cancer patients undergoing chemotherapy. PMID- 12898372 TI - Glomerular diseases in Iranian children: clinico-pathological correlations. AB - Glomerular disease constitutes an important part of renal disease. The purpose of this study was to describe and correlate the types of glomerular diseases reported in native kidney biopsy specimens and their clinical presentations in Iranian children. The medical records and pathological diagnoses of patients with glomerular disease whose native kidney biopsy specimens were received by our hospital during a 10-year period were reviewed. In each patient, pathological diagnosis, clinical presentation, age at onset of disease, gender, and presence of nephrotic proteinuria, hematuria, and hypertension, were determined. Minimal change disease was the most frequent glomerular disease and nephrotic syndrome was the most frequent clinical presentation encountered. For each disease, correlations between the clinical presentation and pathological diagnosis were made. This study describes various glomerular diseases encountered in Iranian children, with their respective clinical presentations. PMID- 12898373 TI - Reverse iontophoresis: a non-invasive technique for measuring blood urea level. AB - Monitoring of the urea level of patients with insufficient kidney function requires repetitive blood sampling. The potentially painful nature of blood sampling and the difficulty of venous access, particularly in premature neonates, as well as possible complications of needle injuries, create many disadvantages. A non-invasive technique needs to be developed for monitoring the urea level for these patients. Reverse iontophoresis has recently gained importance and the possibility of extracting some compounds from body fluids using reverse iontophoresis has been reported in the literature. Moreover, a small, watch-type device has been developed for the determination of blood glucose levels using a similar approach. The aim of the current study was to investigate the possibility of extracting urea from blood through skin using reverse iontophoresis to monitor blood urea levels without taking a blood sample. In vitro iontophoresis studies have indicated that urea may be successfully transferred through the full thickness of human skin. The reverse iontophoresis technique was applied to 17 patients with kidney insufficiency and urea was successfully extracted through their skin into the collection solution. A high correlation ( r(2)=0.878) between urea concentrations in collection solutions and urea levels in the blood was observed. These results suggest that it is possible to make a watch-type device for monitoring blood urea levels. PMID- 12898374 TI - Leptin, soluble leptin receptor, and transforming growth factor-beta1 levels in minimal change nephrotic syndrome. AB - Leptin may contribute to renal pathology in some situations by stimulating transforming growth factor-beta1 (TGF-beta1) synthesis. The soluble leptin receptor (sOb-R) is a transport protein contributing to binding and activation of circulating leptin. We investigated the interaction between serum and urinary leptin, TGF-beta1, and serum sOb-R levels in 38 patients with minimal change nephrotic syndrome (MCNS) aged between 6 and 12 years and 10 age- and sex-matched healthy controls (group III). Patients were divided into two groups: group I, proteinuria exceeding >40 mg/m(2) per hour and group II, patients in remission. Serum leptin levels in group I were significantly lower than those in group II and group III ( P=0.011, P=0.007, respectively). There was a negative correlation between serum leptin levels and proteinuria ( r=-0.52, P=0.02) as well as between serum leptin and sOb-R levels ( r=-0.82, P=0.000) in group I. Urine leptin and sOb-R levels in group I were significantly higher than in group II ( P=0.0021, P=0.001, respectively) and group III ( P=0.07, P=0.009, respectively). Serum TGF beta1 levels in healthy controls (406+/-424 pg/ml) were significantly lower than those in groups I and II ( P=0.004, P=0.000, respectively). However, no significant correlation was found between the serum TGF-beta1 and leptin levels in MCNS patients. In conclusion, low serum leptin, high serum TGF-beta1 and sOb-R levels, and elevated urine leptin concentrations were observed at the onset of MCNS. Since long-term proteinuria and leptinuria might be associated with the progression of renal damage, future in vivo and in vitro studies are needed to explain the interaction between these parameters in different types of nephrotic syndrome. PMID- 12898375 TI - Cyclic voiding urosonography in detecting vesicoureteral reflux in children. AB - Three methods are currently used to identify vesicoureteral reflux (VUR) in children, namely X-ray voiding cystourethrography (VCUG), radionuclide voiding cystography (RVC), and, recently, echo-enhanced voiding urosonography (VUS). It is known that the sensitivity of VCUG and RVC for detecting VUR can be improved by using cyclic procedures, such as repeated bladder filling and voiding. The purpose of our prospective study was to evaluate whether the cyclic procedure is superior to the conventional (one cycle only) procedure in VUS also. VUS was performed in 49 patients, aged 1.4-15.8 years (mean 4.1 years). After the first micturition, the catheter was left in place and the whole procedure was repeated under the same conditions. The results of the first and second cycles and the combined procedure were compared. In the initial cycle, 7 of 35 (20%) refluxing renal units that were detected in the second cycle and 4 of 26 (15%) children with at least unilateral VUR were missed. Cyclic VUS detected 25% more VURs than the conventional (one cycle only) VUS ( P=0.049) and revealed 50% more VUR III than the first cycle. Our results suggest that cyclic VUS is superior to conventional VUS. PMID- 12898376 TI - Effect of the peritoneal dialysis prescription on pentosidine in children. AB - Enhanced formation of advanced glycation end products (AGEs) by peritoneal dialysate containing high dextrose concentrations has been implicated as a source of peritoneal membrane toxicity and loss of viability in patients treated with peritoneal dialysis (PD). The goal of this project was to elucidate the relationship between the structurally defined AGE pentosidine accumulation on peritoneal and plasma proteins and peritoneal membrane function, and to identify clinical factors leading to alterations in these parameters. The study comprised 27 pediatric patients (14 continuous ambulatory PD, 13 chronic cycling PD) on PD for a mean duration of 37.0+/-22.8 months (range 1-120 months) and with a mean age of 13.3+/-4.4 years (range 2.4-20 years). The pentosidine contents of plasma and peritoneal proteins were significantly lower in patients with residual renal function than in patients who were anuric (plasma pentosidine 11.2+/-8.8 vs. 24.1+/-16.6, P=0.02, respectively, peritoneal pentosidine 14.9+/-11.9 vs. 31.1+/ 3.7, P=0.01, respectively). There was no effect of treatment modality on plasma pentosidine (18.1+/-11.2, 18.8+/-19.3, CAPD vs. CCPD, P>0.05) or peritoneal pentosidine content (24.1+/-14.1, 24.9+/-19.6, CAPD vs. CCPD, P>0.05). There was no evidence that increased levels of pentosidine on peritoneal proteins reflect or affect peritoneal membrane function in these patients. Furthermore, there was no effect of peritonitis on the pentosidine content of peritoneal proteins or peritoneal function as measured by peritoneal equilibration test. In conclusion, PD represents a well-tolerated therapy in children with no evidence that current practice causes changes in peritoneal membrane function, or in the peritoneal clearance of plasma or peritoneal proteins rich in pentosidine. PMID- 12898377 TI - Serum selenium level and glutathione peroxidase activity in steroid-sensitive nephrotic syndrome. AB - In our previous study the pattern of glutathione peroxidase (GPX) activity in the course of steroid-sensitive nephrotic syndrome (SSNS) in children suggested a defect in antioxidant defense. In the present report the serum selenium (Se) level, an essential component of GPX activity, was measured in a comparable group of children with SSNS at the same clinical stages at which GPX activity was determined in the previous study. Nephrotic children had normal serum Se levels during the edematous stage, at the end of prednisone treatment, and in remission. At the end of high-dose prednisone treatment, the serum Se level increased ( P<0.01) simultaneously with enhanced activity of GPX. These results suggest that children with SSNS have a persistent defect in the antioxidant defense at the important stage of hydrogen peroxide and fatty acid hydroperoxide decomposition. This defect is transiently alleviated by high-dose prednisone treatment. PMID- 12898378 TI - Long-term outcome of heavy proteinuria in patients under 2 years of age. AB - From January 1985 to July 2000, a retrospective study of 53 patients in Taiwan was performed in order to evaluate the underlying diseases causing heavy proteinuria and the clinical outcome in children under 2 years of age (33 boys and 20 girls). Renal biopsy or autopsy was performed in 26 of the children. Renal pathology revealed 2 patients with congenital nephrosis (CNS) (7.7%), 4 with diffuse mesangial sclerosis (DMS) (15.4%), 4 with minimal change nephrotic syndrome (MCNS) (15.4%), 5 with focal segmental glomerulosclerosis (FSGS) (19.2%), 9 with IgM nephropathy in (34.6%), and 2 with hepatitis B virus associated membranous glomerulonephritis (7.7%). Based on available histology and family history of heavy proteinuria progressing to end-stage renal disease (ESRD), patients were divided into two groups. Group I comprised 10 patients, including CNS (2 cases), DMS (4 cases), and 4 children with a familial history of heavy proteinuria progressing to ESRD. All patients in group I were initially steroid resistant. After methylprednisolone pulse therapy plus cyclosporin A treatment, no patients with CNS or DMS responded, but the other 4 patients experienced a remission. Group II comprised 43 patients; 19 patients (44.2%) were initially steroid resistant. Of these steroid-resistant patients, all experienced remission after methylprednisolone pulse therapy plus cyclosporin A, except 3 children with FSGS. One experienced a thromboembolic event during his clinical course. In conclusion, steroid-resistant nephrotic syndrome (NS) was more common than steroid-sensitive NS in Chinese patients under 2 years of age. Patients with CNS, DMS, or a family history of heavy proteinuria progressing to ESRD had a poor prognosis. Methylprednisolone pulse therapy plus cyclosporin A treatment achieved remission in some children who were initially steroid resistant. This study indicates that children with conditions associated with poor steroid responsiveness (e.g., CNS, DMS) do not respond to immunosuppressive therapy, but other children under 2 years of age, including those with a family history of progression to ESRD, may benefit from aggressive immunosuppressive therapy. PMID- 12898379 TI - Measurement of hydrostatic intraperitoneal pressure: a useful tool for the improvement of dialysis dose prescription. AB - The prescription of peritoneal dialysis should be individualized based on parameters of tolerance and adequacy. Determination of the intraperitoneal fill volume is essential for optimal patient care. Fill volume enhancement is a factor of exchange surface area recruitment: the wetted, contact peritoneal dialysis membrane. Nevertheless, fill volume enhancement can also lead to patient discomfort, with the potential risk of too high an intraperitoneal pressure (hernia, gastro-esophageal reflux). The perception of the individual patient is also a subjective parameter of fill volume tolerance assessment. In contrast, measurement of the hydrostatic intraperitoneal pressure (IPP, cmH(2)O) allows an objective approach to fill volume tolerance. From our clinical experience of more than 10 years of IPP measurements in child care, we can give a recommendation for normal values in children: less than 18 cm of water, usually between 5 and 15 cm, correlated to the intraperitoneal fill volume (naturally), but individually taking into account age, gender, "accustomization" and overall body mass index. PMID- 12898380 TI - The relative importance of sexual reproduction versus clonal spread in an aridland bunchgrass. AB - Festuca idahoensis (Idaho fescue) is a perennial caespitose grass, common in semi arid rangelands of the Intermountain West. To determine how individuals are recruited into a population, we studied two long-term monitoring plots that were established in 1937 at the Northern Great Basin Experimental Range in southeastern Oregon. The plots measured 3.05x3.05 m, and were located approximately 30 m apart. One plot was ungrazed, the other was subject to moderate levels of cattle grazing. The number of F. idahoensis plants in both plots increased ten-fold between 1937 and 1996, but whether this was due primarily to reproduction by seed or clonal fragmentation was unknown. In 1996, we mapped and sampled 160 plants of F. idahoensis. We used dominant inter-simple sequence repeat (ISSR) markers and codominant allozyme markers in order to identify genetic individuals and measure genetic diversity. Both plots were characterized by high levels of genetic and clonal diversity. When information from ISSRs, allozymes and sample location were combined, 126 genets were recognized, each consisting of one to four samples (ramets). By measuring the diameter of clones surrounding plants that were present in 1937, we estimated that clonal spread occurred at a rate of approximately 3.7 cm per decade, and thus was of secondary importance in the maintenance and increase of F. idahoensis stands. Sexual reproduction, rather than clonal fragmentation, accounted for most of the recruitment of new plants into these plots. The grazed plot had fewer ramets, genotypes, and clones than the ungrazed plot, but the ramets were significantly larger. Levels of genetic diversity did not differ in the grazed and ungrazed plots, but there was some evidence for a small, but significant level of genetic differentiation between the two. The results also indicate that F. idahoensis has the potential to be a long-lived species with some individuals persisting in excess of 60 years. This study demonstrates how long-term monitoring can be supplemented by genetic analysis to obtain detailed information on the population dynamics of plants. In the case of this community dominant species, this provides essential information for understanding succession and developing management and restoration strategies. PMID- 12898381 TI - Macroecology of habitat choice in long-distance migratory birds. AB - Patterns of habitat choice in ecological communities are not only influenced by present-day selective forces but also by historical processes, such as the biogeographical history of the lineages they are composed of. Nevertheless, it has been very difficult to test historical factors. The possible tropical origin of long-distance migratory birds provides an opportunity for such a test. If habitat choice of long-distance migrants is inherited from their tropical ancestors then Nearctic long-distance migrants might have acquired their habitat choice from Neotropical forest species and European long-distance migrants from African savannah species. Here we use a macroecological approach to show that this hypothesis can be confirmed. Long-distance migrants in the Nearctic are found in forested habitat types, while those in Europe are found in open ones. In comparison, the habitat choice of residents and short-distance migrants (in genera without long-distance migration) does not differ between the Nearctic and Europe. These results demonstrate that habitat choice in temperate bird communities can be explained by the tropical history of long-distance migrants. Thus, habitat choice seems to be shaped not only by local mechanisms, but also by processes acting on much larger spatial and temporal scales. PMID- 12898382 TI - Elevated CO2 reduces sap flux in mature deciduous forest trees. AB - We enriched in CO2 the canopy of 14 broad-leaved trees in a species-rich, ca. 30 m-tall forest in NW Switzerland to test whether elevated CO2 reduces water use in mature forest trees. Measurements of sap flux density (JS) were made prior to CO2 enrichment (summer 2000) and throughout the first whole growing season of CO2 exposure (2001) using the constant heat-flow technique. The short-term responses of sap flux to brief (1.5-3 h) interruptions of CO2 enrichment were also examined. There were no significant a priori differences in morphological and physiological traits between trees which were later exposed to elevated CO2 (n=14) and trees later used as controls (n=19). Over the entire growing season, CO2 enrichment resulted in an average 10.7% reduction in mean daily JS across all species compared to control trees. Responses were most pronounced in Carpinus, Acer, Prunus and Tilia, smaller in Quercus and close to zero in Fagus trees. The JS of treated trees significantly increased by 7% upon transient exposure to ambient CO2 concentrations at noon. Hence, responses of the different species were, in the short term, similar in magnitude to those observed over the whole season (though opposite because of the reversed treatment). The reductions in mean JS of CO2-enriched trees were high (22%) under conditions of low evaporative demand (vapour pressure deficit, VPD <5 hPa) and small (2%) when mean daily VPD was greater than 10 hPa. During a relatively dry period, the effect of elevated CO2 on JS even appeared to be reversed. These results suggest that daily water savings by CO2-enriched trees may have accumulated to a significantly improved water status by the time when control trees were short of soil moisture. Our data indicate that the magnitude of CO2 effects on stand transpiration will depend on rainfall regimes and the relative abundance of the different species, being more pronounced under humid conditions and in stands dominated by species such as Carpinus and negligible in mono-specific Fagus forests. PMID- 12898383 TI - Foliar quality influences tree-herbivore-parasitoid interactions: effects of elevated CO2, O3, and plant genotype. AB - This study examined the effects of carbon dioxide (CO2)-, ozone (O3)-, and genotype-mediated changes in quaking aspen (Populus tremuloides) chemistry on performance of the forest tent caterpillar (Malacosoma disstria) and its dipteran parasitoid (Compsilura concinnata) at the Aspen Free-Air CO2 Enrichment (FACE) site. Parasitized and non-parasitized forest tent caterpillars were reared on two aspen genotypes under elevated levels of CO2 and O3, alone and in combination. Foliage was collected for determination of the chemical composition of leaves fed upon by forest tent caterpillars during the period of endoparasitoid larval development. Elevated CO2 decreased nitrogen levels but had no effect on concentrations of carbon-based compounds. In contrast, elevated O3 decreased nitrogen and phenolic glycoside levels, but increased concentrations of starch and condensed tannins. Foliar chemistry also differed between aspen genotypes. CO2, O3, genotype, and their interactions altered forest tent caterpillar performance, and differentially so between sexes. In general, enriched CO2 had little effect on forest tent caterpillar performance under ambient O3, but reduced performance (for insects on one aspen genotype) under elevated O3. Conversely, elevated O3 improved forest tent caterpillar performance under ambient, but not elevated, CO2. Parasitoid larval survivorship decreased under elevated O3, depending upon levels of CO2 and aspen genotype. Additionally, larval performance and masses of mature female parasitoids differed between aspen genotypes. These results suggest that host-parasitoid interactions in forest systems may be altered by atmospheric conditions anticipated for the future, and that the degree of change may be influenced by plant genotype. PMID- 12898384 TI - Successional changes of Collembola and soil microbiota during forest rotation. AB - Dynamic approaches to forest ecosystems are surprisingly rare. Here we report about successional changes in collembolan community structure and microbial performances during forest rotation. The study was carried out in a chronosequence of four spruce forest stands (5-, 25-, 45-, and 95 years old; Tharandter forest, Germany). CO2 release significantly increased after clear cutting and the amount of C stored in the organic layer subsequently declined. The early phase of forest rotation was characterized by a very active decomposer microflora, stimulation of both fungi and bacteria as well as by a high abundance of surface-oriented Collembola. In addition, collembolan species turnover was accelerated. While the biomass of fungi further increased at intermediate stages of forest rotation, the metabolic activity of the microflora was low, the functional diversity of bacteria declined and the collembolan community became impoverished. Euedaphic species dominated during this stage of forest development. These changes can be explained by both reduction in microhabitat diversity and depletion of food sources associated with an accumulation of recalcitrant soil organic matter. Results of the General Regression Model procedure indicate a shift from specific associations between collembolan functional groups and microbiota at the early stage of forest rotation to a more diffuse pattern at intermediate stages. Though the hypothesis that Collembola are relatively responsive to changes in environmental conditions is confirmed, consistently high community similarity suggests a remarkable persistence of some components of microarthropod assemblages. Our study provides evidence for substantial ecosystem-level implications of changes in the soil food web during forest rotation. Moreover, correlations between bacterial parameters and Collembola point to the overarching impact of differences in the composition of the microbial community on microarthropods. PMID- 12898385 TI - Prevalence of coccidia parasites (Protozoa) in red squirrels (Sciurus vulgaris): effects of host phenotype and environmental factors. AB - We investigated the relative importance of environmental factors versus host phenotype in determining parasite prevalence in Eurasian red squirrels (Sciurus vulgaris). One hundred and forty-three fecal samples of 116 different squirrels collected in 2000 and 2001 from five study areas in the Italian Alps, were examined for intestinal protozoans. Two species of Eimeria were present with a medium to high prevalence in both years and in all areas, while two other species were rare, occurring only in some areas and not in all years. Cryptosporidium parvum had a high prevalence in the two study areas of the Western Alps, while in the three areas of the Central Alps it was recorded only once. The prevalence of Eimeria sciurorum and C. parvum fluctuated in parallel with squirrel density, suggesting a possible correlation between the presence of these protozoans and host density. A gender effect on E. sciurorum prevalence at low density could be explained by different space use patterns and social organization of males and females. C. parvum occurred more frequently in young squirrels, suggesting an acquired immunity in adults, but age-related susceptibility was not found for eimerian species. The coccidian community was more similar within than between regions, and study area and year were key parameters in predicting coccidia infection. There was no evidence of competition between coccidian species, but one positive interaction between E. sciurorum and E. andrewsi was observed. Our results suggest that the effects of geographic region, area features, and year effects probably related to fluctuations in host population density, were more important than individual phenotypic host characteristics in structuring the coccidian assemblage and determining levels of parasite prevalence in red squirrel populations. PMID- 12898386 TI - Disentangling a rainforest food web using stable isotopes: dietary diversity in a species-rich ant community. AB - For diverse communities of omnivorous insects such as ants, the extent of direct consumption of plant-derived resources vs. predation is largely unknown. However, determination of the extent of "herbivory" among ants may be crucial to understand the hyper-dominance of ants in tropical tree crowns, where prey organisms tend to occur scarcely and unpredictably. We therefore examined N and C stable isotope ratios (delta(15)N and delta(13)C) in 50 ant species and associated insects and plants from a tropical rainforest in North Queensland, Australia. Variation between ant species was pronounced (range of species means: 7.1 per thousand in delta(15)N and 6.8 per thousand in delta(13)C). Isotope signatures of the entire ant community overlapped with those of several herbivorous as well as predacious arthropods. Variability in delta(15)N between ants was not correlated with plant delta(15)N from which they were collected. Ant species spread out in a continuum between largely herbivorous and purely predacious taxa, with a high degree of omnivory. Ant species' delta(15)N were consistent with the trophic level predicted by natural feeding observations, but not their delta(13)C. Low delta(15)N levels were recorded for ant species that commonly forage for nectar on understorey or canopy plants, intermediate levels for species with large colonies that were highly abundant on nectar and honeydew sources and were predacious, and the highest levels for predominantly predatory ground-foraging species. Colonies of the dominant weaver-ants (Oecophylla smaragdina) had significantly lower delta(15)N in mature forests (where preferred honeydew and nectar sources are abundant) than in open secondary vegetation. N concentration of ant dry mass showed only very limited variability across species and no correlation with trophic levels. This study demonstrates that stable isotopes provide a powerful tool for quantitative analyses of trophic niche partitioning and plasticity in complex and diverse tropical omnivore communities. PMID- 12898387 TI - Expression of metabotropic glutamate receptor group I in rat gustatory papillae. AB - Taste-metabotropic glutamate receptor 4 (taste-mGluR4) and the heteromers of T1R1 and T1R3 are candidate receptors involved in the sense of umami (monosodium glutamate) taste. Although the expression of group III mGluRs (taste-mGluR4) has been demonstrated in taste tissues, no mention has been made of the expression of group I mGluRs (mGluR1 and mGluR5) in taste tissues. We examined the expression of mGluR1 and mGluR5 in rat gustatory tissues by using reverse transcription polymerase chain reaction (RT-PCR), in situ hybridization, immunohistochemistry and immunoelectron microscopy. RT-PCR assay showed that mGluR1 alpha and mGluR1 beta mRNAs were expressed in circumvallate papillae, but mGluR5 mRNA was not expressed. The positive signals of mGluR1 mRNA were detected only in circumvallate taste buds by in situ hybridization analysis. In cryosections of fungiform, foliate and circumvallate papillae, the antibody against mGluR1 alpha gave intense labeling on the taste hairs in all taste pores examined. In the developing taste buds, the positive signals of mGluR1alpha in taste hairs gradually increased with the increase in number of taste bud cells. These results show that, in addition to taste-mGluR4 and the heteromer of T1R1 and T1R3, mGluR1 alpha may function as a receptor for glutamate (umami) taste sensation. PMID- 12898388 TI - A degeneracy study in the circulant and bordered-circulant approach to birdcage and planar coils. AB - A method for finding closed-form solutions for the normal mode frequencies of systems with circulant (2 pi/n) symmetry was investigated. This method is particularly useful for questions of degeneracy that arise when one considers parallel imaging techniques like SENSE and SMASH in MRI. It is applicable to systems that include birdcage coils as well as planar coils with the appropriate rotational symmetry. A proof is given that complete degeneracy of all normal mode frequencies is impossible when all mutual inductive couplings are included. We tested the method against measurements made on a planar coil array and on an 8 element birdcage coil. The inclusion of the co-rotating end-ring mode changes the fundamental symmetry of the system from circulant to 'bordered circulant.' Closed form solutions for the normal mode frequencies of a bordered circulant system are also given. PMID- 12898389 TI - Metabolic engineering of Escherichia coli for improved 6-deoxyerythronolide B production. AB - Escherichia coli is an attractive candidate as a host for polyketide production and has been engineered to produce the erythromycin precursor polyketide 6 deoxyerythronolide B (6dEB). In order to identify and optimize parameters that affect polyketide production in engineered E. coli, we first investigated the supply of the extender unit ( 2S)-methylmalonyl-CoA via three independent pathways. Expression of the Streptomyces coelicolor malonyl/methylmalonyl-CoA ligase ( matB) pathway in E. coli together with methylmalonate feeding resulted in the accumulation of intracellular methylmalonyl-CoA to as much as 90% of the acyl-CoA pool. Surprisingly, the methylmalonyl-CoA generated from the matB pathway was not converted into 6dEB. In strains expressing either the S. coelicolor propionyl-CoA carboxylase (PCC) pathway or the Propionibacteria shermanii methylmalonyl-CoA mutase/epimerase pathway, methylmalonyl-CoA accumulated up to 30% of the total acyl-CoA pools, and 6dEB was produced; titers were fivefold higher when strains contained the PCC pathway rather than the mutase pathway. When the PCC and mutase pathways were expressed simultaneously, the PCC pathway predominated, as indicated by greater flux of (13)C-propionate into 6dEB through the PCC pathway. To further optimize the E. coli production strain, we improved 6dEB titers by integrating the PCC and mutase pathways into the E. coli chromosome and by expressing the 6-deoxyerythronolide B synthase (DEBS) genes from a stable plasmid system. PMID- 12898390 TI - Harmful algal blooms: causes, impacts and detection. AB - Blooms of autotrophic algae and some heterotrophic protists are increasingly frequent in coastal waters around the world and are collectively grouped as harmful algal blooms (HABs). Blooms of these organisms are attributed to two primary factors: natural processes such as circulation, upwelling relaxation, and river flow; and, anthropogenic loadings leading to eutrophication. Unfortunately, the latter is commonly assumed to be the primary cause of all blooms, which is not the case in many instances. Moreover, although it is generally acknowledged that occurrences of these phenomena are increasing throughout the world's oceans, the reasons for this apparent increase remain debated and include not only eutrophication but increased observation efforts in coastal zones of the world. There is a rapidly advancing monitoring effort resulting from the perception of increased impacts from these HABs, manifested as expanding routine coastal monitoring programs, rapid development and deployment of new detection methods for individual species, toxins, and toxicities, and expansion of coastal modeling activities towards observational forecasts of bloom landfall and eventually bloom prediction. Together, these many efforts will provide resource managers with the tools needed to develop effective strategies for the management and mitigation of HABs and their frequently devastating impacts on the coastal environment. PMID- 12898391 TI - Microbial characterization and hydrocarbon biodegradation potential of natural bilge waste microflora. AB - Shipping operations produce oily wastes that must be managed properly to avoid environmental pollution. The aim of this study was to characterize microorganisms occurring in ship bilge wastes placed in open lagoons and, particularly, to assess their potential to degrade polycyclic aromatic hydrocarbons (PAHs). A first-order kinetic was suitable for describing hydrocarbon biodegradation after 17 days of treatment. The calculated rate constants were 0.0668 and 0.0513 day( 1) with a corresponding half-life of 10.3 and 13.5 days for the aliphatic and aromatic hydrocarbon fractions, respectively. At day 17, PAH removal percentages were: acenaphtylene 100, fluorene 95.2, phenanthrene 93.6, anthracene 70.3, and pyrene 71.5. Methyl phenanthrene removals were lower than that of their parent compound (3-methyl phenanthrene 83.6, 2-methyl phenanthrene 80.8, 1-methyl phenanthrene 77.3, 9-methyl phenanthrene 75.1, and 2,7-dimethyl phenanthrene 76.6). Neither pure cultures nor the microbial community from these wastes showed extracellular biosurfactant production suggesting that the addition of an exogenously produced biosurfactant may be important in enhancing hydrocarbon bioavailability and biodegradation. DNA analysis of bilge waste samples revealed a ubiquitous distribution of the nahAc genotype in the dump pools. Although almost all of the isolates grew on naphthalene as sole carbon source, only some of them yielded nahAc amplification under the experimental conditions used. The variety of PAHs in bilge wastes could support bacteria with multiple degradation pathways and a diversity of catabolic genes divergent from the classical nah-like type. PMID- 12898392 TI - Changes in the rate of photosynthesis accompanying the yield increase in wheat cultivars released in the past 50 years. AB - Photosynthetic rates in different development stages were carefully investigated in 18 cultivars of winter wheat released in the period between 1945 and 1995 in the area of Beijing, China. During this period, the recorded grain yield has increased eightfold. However, when those cultivars were planted and managed in the same environment, the difference was reduced to only 36%, indicating that agronomic practices are the most important factors for grain yield. Agronomic features have changed greatly in the past 50 years, through increasing the harvest index (R2 = 0.89, P < 0.05), shortening plant height (R2 = 0.77, P < 0.05) and slightly increasing flag leaf areas (R2 = 0.45, P < 0.05), which is mostly in agreement with many other researchers. In contrast to many reports, however, this study found a genetic increase in the rate of photosynthesis per unit leaf area. From the mid-stem elongation to soft dough stages, the average photosynthetic rates at saturated photosynthetic photon flux density (P(sat)) increased by 44%. In the process, the stomatal conductance (g(s)) also increased by 122%. Grain yield was positively related to the mean values of P(sat) (R2 = 0.61, P < 0.01) and g(s) (R2 = 0.67, P < 0.01) in the six development stages. Our experiment may suggest that increase in grain yield was associated with the elevation of leaf photosynthetic rate and stomatal conductance over the past 50 years. PMID- 12898393 TI - Multicentric glioma: our experience in 25 patients and critical review of the literature. AB - Multicentric gliomas are interesting and well-recognised entities with a yet unknown rate of occurrence. Single cases or small series are reported in the literature accessible to us, and we think this is the first large series describing true multicentric gliomas. We reviewed 25 patients selected according to the criteria defined by Batzdorf and Malamud. Multicentricity was found in 2% of patients with malignant gliomas. Longer survival was observed in patients who underwent surgical excision of the multicentric lesions. Multicentric tumours are rare clinical entities. Our data suggest that they should be surgically removed whenever possible, and histopathologic examination of the lesions is always advisable if they are located in sites inaccessible to surgery. Stereotactic biopsy represents a safe and satisfactory method for achieving sure diagnosis. PMID- 12898394 TI - Global nutritional profiling for mutant and chemical mode-of-action analysis in filamentous fungi. AB - We describe a method for gene function discovery and chemical mode-of-action analysis via nutrient utilization using a high throughput Nutritional Profiling platform suitable for filamentous microorganisms. We have optimized the growth conditions for each fungal species to produce reproducible optical density growth measurements in microtiter plates. We validated the Nutritional Profiling platform using a nitrogen source utilization assay to analyze 21 Aspergillus nidulans strains with mutations in the master nitrogen regulatory gene, areA. Analysis of these data accurately reproduced expected results and provided new data to demonstrate that this platform is suitable for fine level phenotyping of filamentous fungi. Next, we analyzed the differential responses of two fungal species to a glutamine synthetase inhibitor, illustrating chemical mode-of-action analysis. Finally, a comparative phenotypic study was performed to characterize carbon catabolite repression in four fungal species using a carbon source utilization assay. The results demonstrate differentiation between two Aspergillus species and two diverse plant pathogens and provide a wealth of new data on fungal nutrient utilization. Thus, these assays can be used for gene function and chemical mode-of-action analysis at the whole organism level as well as interspecies comparisons in a variety of filamentous fungi. Additionally, because uniform distribution of growth within wells is maintained, comparisons between yeast and filamentous forms of a single organism can be performed. PMID- 12898395 TI - Learning from yeasts: intracellular sensing of stress conditions. AB - One intriguing challenge in modern biology is to understand how cells respond to, and distinguish between different stressing stimuli. Evidence accumulated in recent years indicates that a network of signaling pathways extends from the plasma membrane to the very core of the cell nucleus to transduce environmental changes into a graded transcriptional response. Although many steps still remain unclear, studies on the stress-activated protein kinase (SAPK) pathways and related mechanisms provide insight into the biochemistry that regulates signal transmission and leads to outcomes such as cell adaptation and differentiation. This review focuses on selected topics of current interest related to the sensing of stress signals in cells of the fission yeast Schizosaccharomyces pombe. Because signaling pathways appear to be evolutionarily well conserved, yeasts may be useful models to learn how higher eukaryotes sense and respond to stresses at the cellular level. PMID- 12898397 TI - Reflections on the classification of yeasts for different end-users in biotechnology, ecology, and medicine. AB - The approach to yeast identification has significantly changed in just a few decades due to the rapid increase in basic biological knowledge, increased interest in the practical applications and biodiversity of this important microbial group, and enormous technological advances. While some conventional methods can still be validly applied, many molecular techniques have been developed that allow for strain classification on all taxonomic levels. A critical evaluation of the actual scope of each identification procedure will in the end determine the most appropriate use of the many protocols now available. Nonetheless, the oldest tool of microbiology, the microscope, is still a fundamental accessory for studies involving yeast biology, biodiversity and taxonomy. PMID- 12898396 TI - Phaffia rhodozyma: colorful odyssey. AB - Phaffia rhodozyma was isolated by Herman Phaff in the 1960s, during his pioneering studies of yeast ecology. Initially, the yeast was isolated from limited geographical regions, but isolates were subsequently obtained from Russia, Chile, Finland, and the United States. The biological diversity of the yeast is more extensive than originally envisioned by Phaff and his collaborators, and at least two species appear to exist, including the anamorph Phaffia rhodozyma and the teleomorph Xanthophyllomyces dendrorhous. The yeast has attracted considerable biotechnological interest because of its ability to synthesize the economically important carotenoid astaxanthin (3,3'-dihydroxy beta, beta-carotene-4,4'-dione) as its major pigment. This property has stimulated research on the biology of the yeast as well as development of the yeast as an industrial microorganism for astaxanthin production by fermentation. Our laboratory has isolated several mutants of the yeast affected in carotenogenesis, giving colonies a vivid array of pigmentation. We have found that nutritional and environmental conditions regulate astaxanthin biosynthesis in the yeast, and have demonstrated that astaxanthin protects P. rhodozyma from damage by reactive oxygen species. We proposed in the 1970s that P. rhodozyma could serve as an economically important pigment source in animal diets including salmonids, lobsters, and the egg yolks of chickens and quail, in order to impart characteristic and desirable colors. Although P. rhodozyma/Xanthomyces dendrorhous has been studied by various researchers for nearly 30 years, it still attracts interest from yeast biologists and biotechnologists. There is a bright and colorful outlook for P. rhodozyma/X. dendrorhous from fundamental and applied research perspectives. PMID- 12898398 TI - Comparative genomics of yeast species: new insights into their biology. AB - The genomes of two hemiascomycetous yeasts (Saccharomyces cerevisiae and Candida albicans) and one archiascomycete (Schizosaccharomyces pombe) have been completely sequenced and the genes have been annotated. In addition, the genomes of 13 more Hemiascomycetes have been partially sequenced. The amount of data thus obtained provides information on the evolutionary relationships between yeast species. In addition, the differential genetic characteristics of the microorganisms explain a number of distinctive biological traits. Gene order conservation is observed between phylogenetically close species and is lost in distantly related species, probably due to rearrangements of short regions of DNA. However, gene function is much more conserved along evolution. Compared to S. cerevisiae and S. pombe, C. albicans has a larger number of specific genes, i.e., genes not found in other organisms, a fact that can account for the biological characteristics of this pathogenic dimorphic yeast which is able to colonize a large variety of environments. PMID- 12898399 TI - Fungal biotechnology. AB - Fungi are used in many industrial processes, such as the production of enzymes, vitamins, polysaccharides, polyhydric alcohols, pigments, lipids, and glycolipids. Some of these products are produced commercially while others are potentially valuable in biotechnology. Fungal secondary metabolites are extremely important to our health and nutrition and have tremendous economic impact. In addition to the multiple reaction sequences of fermentations, fungi are extremely useful in carrying out biotransformation processes. These are becoming essential to the fine-chemical industry in the production of single-isomer intermediates. Recombinant DNA technology, which includes yeasts and other fungi as hosts, has markedly increased markets for microbial enzymes. Molecular manipulations have been added to mutational techniques as a means of increasing titers and yields of microbial processes and in the discovery of new drugs. Today, fungal biology is a major participant in global industry. Moreover, the best is yet to come as genomes of additional species are sequenced at some level (cDNA, complete genomes, expressed sequence tags) and gene and protein arrays become available. PMID- 12898400 TI - Molecular evolution in yeast of biotechnological interest. AB - The importance of yeast in the food and beverage industries was only realized about 1860, when the role of these organisms in food manufacture became evident. Since they grow on a wide range of substrates and can tolerate extreme physicochemical conditions, yeasts, especially the genera Saccharomyces and Kluyveromyces, have been applied to many industrial processes, Industrial strains of these genera are highly specialized organisms that have evolved to utilize a range of environments and ecological niches to their full potential. This adaptation is called "domestication". This review describes the phylogenetic relationships among Saccharomyces and Kluyveromyces species and the different mechanisms involved in the adaptive evolution of industrial yeast strains. PMID- 12898402 TI - [The limits of refractive surgery]. PMID- 12898401 TI - Biotechnology of natural and winery-associated strains of Saccharomyces cerevisiae. AB - A new body of evidence challenges the original consolidated theory of Pasteur on the natural (vineyard) origin of wine strains of Saccharomyces cerevisiae and instead indicates a local, winery-restricted life cycle. The findings open novel biotechnological perspectives for obtaining autochthonous selected starters for the wine industry. A local, individual, and specific fermenting yeast flora, mass selected year after year through many generations of S. cerevisiae in grape must, is present on the surfaces of every winery. These yeast strains are endowed with exceptional enological properties and capable of producing an assortment of volatile compounds apparently contributing to the specific bouquet of locally produced wines. PMID- 12898403 TI - [Understanding trabecular meshwork function will improve glaucoma treatment]. PMID- 12898404 TI - [Phacotrabeculectomy: one site versus two site approach]. PMID- 12898405 TI - [Intracellular pathways leading to apoptosis of retinal cells]. AB - Some ocular diseases, such as retinitis pigmentosa, age-related macular degeneration, glaucoma, retinal detachment, diabetic retinopathy or pathological myopia result in apoptotic death of retinal cells. Caspases are proteases that take part in the intracellular signaling pathways that cause apoptosis in different cell types. By using different experimental models, an increase in caspase activity in retinal cells after damage has been shown. However, some studies have shown that other molecules apart from caspases are activated during apoptosis in the retina. Moreover, the type of caspase that is activated seems to be different depending on the type of retinal cell and the pathology analysed. Inhibition of the processes that result in retinal cell apoptosis could decrease the number of dead cells and prevent the irreversible loss of visual function associated with some pathologies such as glaucoma. In this study we aim to review the basic research studies published in the last years in order to know the molecules and pathways involved in the processes which cause apoptosis of the different types of retinal cells. PMID- 12898406 TI - [Phthiriasis palpebrarum: diagnosis and treatment]. AB - Phthiriasis palpebrarum, caused by Phthirus pubis, is an uncommon cause of blepharoconjunctivitis; therefore, this condition is easily misdiagnosed. When diagnosed, genital involvement must be ruled out. Association with other venereal diseases is common. Affected children must be searched for sexual abuse. The number of diagnosed patients in our department has increased in recent years. We review the epidemiological, clinical and diagnostic features of phthriasis palpebrarum as well as the different treatment options to eradicate the parasite and to prevent infestations. PMID- 12898407 TI - [Normal human conjunctival epithelium expresses MUC13, MUC15, MUC16 and MUC17 mucin genes]. AB - PURPOSE: The ocular surface epithelia express at least five mucin genes of the total of 17 human mucin genes that have been identified so far. This study was designed to determine the expression profile of mucin genes in conjunctival impression cytology (CIC) samples from healthy subjects. METHODS: Two polyethersulfone filters were applied to the superior conjunctiva of both eyes from eight healthy donors. Polymerase chain reaction (PCR) was performed using isolated and retrotranscripted total RNA obtained from the CIC samples to study the expression of all known human mucin genes. Following amplification, PCR products were electrophoresed on 1.5% agarose gel and stained with ethidium bromide to confirm that only a single band was obtained when amplifying all cDNAs with the convenient primers. RESULTS: Transcripts of the previously reported conjunctival mucin genes MUC1, MUC2, MUC4, MUC5AC, and MUC7 were detected in all samples. In addition, transcripts of MUC13, MUC15, MUC16 and MUC17 mucin genes also were detected. Amplified products by conventional PCR showed the expected amplicon size. Transcripts of MUC3A, MUC3B, MUC5B, MUC6, MUC8, MUC11, and MUC12 mucin genes were not detected. CONCLUSION: The expression of four additional mucin mRNA (MUC13, MUC15, MUC16, and MUC17) has been proved in human conjunctival epithelium from healthy donors for the first time. The function of these genes remains to be further elucidated. PMID- 12898408 TI - [Prevalence and risk factors of pseudoexfoliation syndrome in institutionalized geriatric patients in Navarra]. AB - PURPOSE: To establish the prevalence of pseudoexfoliation syndrome (PSX) in an institutionalized geriatric population in Navarra. To study the risk factors for the development of this disease. MATERIAL AND METHODS: 268 nursing home residents were studied, with a mean age of 81 years. The presence of PSX material in the anterior segment was best appreciated by slit lamp after pupillary dilation. We assessed its association with 13 ocular factors and 14 systemic factors. RESULTS: We found 10.1% (27) of subjects with PSX, 19 cases were unilateral and 8 bilateral. The frequency detected in subjects from Navarra was 7.9% versus 21.2% in subjects from other regions (p=0.02), although the significance was lost after multivariant logistic regression. The PSX was 3.5 times more frequent in patients suffering heart failure (p=0.01). The PSX was associated with anti-glaucomatous treatment, which multiplies the risk by 3.2 times (p=0.02); patients affected with age-related geographic macular atrophy had a 2.6 fold increased frequency (p=0.03), both after adjusting for age remained significant. We did not find any association between the PSX and other systemic factors (systemic hypertension and diabetes mellitus) or ocular factors (senile arc and cataract). CONCLUSIONS: The prevalence of PSX syndrome seems to be lower in an institutionalized population in Navarra than in other Spanish regions. The correlation with heart failure and age-related geographic macular atrophy suggests the possibility of a vascular role. The epidemiological association with increased IOP is confirmed. PMID- 12898409 TI - [Juvenile Diabetic cataract. A rare finding which lead us to the diagnosis of this illness]. AB - CASE REPORT: A 13-year-old female presented as an emergency, complaining of loss of vision during the last three months. On examination she had bilateral posterior subcapsular cataracts with snowflake opacities and a blood glucose of 731 mg/dl with ketoacidosis. DISCUSSION: True diabetic cataracts should be differentiated from other lens opacities in diabetics. The latter, identical to senile cataracts, are very common but appear earlier in diabetic patients and are not considered true diabetic cataracts (which are rare). Although true diabetic cataracts are infrequent, the appearance of lens opacities in young patients should alert the ophthalmologist to the existence of diabetes mellitus. PMID- 12898410 TI - [The otolaryngologic, neurologic and ophthalmologic movement in Europe]. PMID- 12898411 TI - [Some pictures by Hermenegildo Arruga]. PMID- 12898412 TI - Phytoestrogens: recent developments. AB - Phytoestrogens are polyphenolic non-steroidal plant compounds with estrogen-like biological activity. Based on their chemical structure, phytoestrogens can be classified into four main groups, i. e., isoflavonoids, flavonoids, stilbenes, and lignans. For each group, the chemistry, dietary sources and biotransformation of the most interesting compounds will be discussed. Since phytoestrogens are structurally very similar to the estrogen 17beta-estradiol, they may exhibit selective estrogen receptor modulating activities. Therefore, special attention will be given to the hormonal effects of various isoflavonoids, including genistein, daidzein, coumestrol and equol, several prenylated flavonoids, especially 8-prenylnaringenin, and the stilbene resveratrol. Furthermore, their non-hormonal effects will be discussed briefly. Finally, the latest developments on the potential protective properties of phytoestrogens and phytoestrogen containing foods against hormone-dependent breast and prostate cancers and cardiovascular diseases, and as estrogen replacement therapy for postmenopausal women will be discussed. PMID- 12898413 TI - Immunomodulatory activities of flavonoids, monoterpenoids, triterpenoids, iridoid glycosides and phenolic compounds of Plantago species. AB - A number of Plantago spp. especially P. major has long been used in the treatment of diseases such as infection, inflammation and cancer. In this study, we evaluated the immunomodulatory activities of five chemical classes of pure compounds obtained from the Plantago genus on human peripheral blood mononuclear cells (PBMC). Studies were conducted on lymphocyte transformation by BrdU immunoassay and secretion of interferon-gamma (IFN-gamma) using an ELISA assay. Results showed that the water-soluble compounds, namely aucubin, chlorogenic acid, ferulic acid, p-coumaric acid and vanillic acid, enhanced the activity of human lymphocyte proliferation and secretion of IFN-gamma. Among the water insoluble compounds, with the exception of luteolin, both baicalein and baicalin showed an enhancement of the human PBMC. Although oleanolic acid and ursolic acid of the triterpenoids did not significantly affect the proliferation of PBMC, they exhibited a strong stimulation of IFN-gamma secretion. Linalool, a monoterpenoid, showed a similar immunomodulatory activity as the triterpenoids. The present study concludes that the tested compounds, which possess immunostimulating activities, may contribute to the traditional claims of Plantago-based natural products used in treating cancers and infectious diseases. PMID- 12898414 TI - Effects of plumbagin on platelet aggregation and platelet-neutrophil interactions. AB - The effects of plumbagin were investigated on platelet aggregation in vitro and ex vivo, on the binding of thrombin-stimulated platelets to neutrophils, and platelet aggregation induced by intact neutrophils and N-formyl-methionyl-leucyl phenylalanine (fMLP) or platelet activating factor (PAF) activated neutrophils, by use of the methods of Hamburger, McEver and Born, respectively. The results showed that plumbagin in vitro significantly inhibited adenosine diphosphate (ADP)-, arachidonic acid (AA)-, or platelet activating factor (PAF)-induced platelet aggregation, in a concentration-dependent manner. The medium inhibitory concentrations (IC 50 ) were 39.4, 82.7 and 38.1 microM, respectively. Intragastric plumbagin at 10 mg/kg markedly suppressed platelet aggregation induced by ADP, AA, or PAF. Plumbagin decreased the binding between thrombin stimulated platelets and neutrophils with an IC 50 of 62.9 microM. Plumbagin significantly inhibited washed platelet aggregation stimulated by fMLP- or PAF activated neutrophils. The IC 50 values were 54.3 and 47.6 microM, respectively. On the other hand, plumbagin and aspirin increased the inhibition of intact neutrophils on AA-induced platelet aggregation. It is suggested that plumbagin inhibited platelet aggregation in vitro and ex vivo, suppressed the binding of activated platelets to neutrophils, inhibited platelet aggregation induced by activated neutrophils, and increased inhibition of intact neutrophils on platelet reactivity. Abbreviations. DMSO:dimethyl sulphoxide fMLP: N-formyl-methionyl leucyl-phenylalanine ADP:adenosine diphosphate AA:arachidonic acid PAF:platelet activating factor PMID- 12898416 TI - Anti-angiogenic and anti-tumor activities of isoflavonoids from the rhizomes of Belamcanda chinensis. AB - The present study was carried out to clarify whether tectorigenin and tectoridin isolated from the rhizomes of Belamcanda chinensis (Iridaceae) inhibit angiogenesis by the experimental methods in vitro and in vivo. Tectorigenin and tectoridin decreased angiogenesis of both chick embryos in the chorioallantoic membrane assay and basic fibroblast growth factor-induced vessel formation in the mouse Matrigel plug assay. Both compounds also reduced the proliferation of calf pulmonary arterial endothelial (CPAE) cells and found to possess relatively weak gelatinase/collagenase inhibitory activity in vitro. Tectorigenin exhibited a much stronger anti-proliferative activity than its glycoside, tectoridin and was almost equipotent to that of genistein, a reference drug. Tectorigenin, when administered subcutaneously at the dose of 30 mg/kg for 20 days to mice implanted with murine Lewis lung carcinoma (LLC), caused a significant inhibition of tumor volume by 30.8 %. Tectorigenin and tectoridin, when treated i. p. at the same dosage for 10 days to ICR mice bearing sarcoma 180, caused a significant suppression in tumor weight by 44.2 and 24.8 %, respectively. PMID- 12898415 TI - In vivo anti-inflammatory and antinociceptive effects of liriodendrin isolated from the stem bark of Acanthopanax senticosus. AB - In the present study, liriodendrin isolated by activity-guided fractionation from the ethyl acetate (EtOAc) extracts of the stem bark of Acanthopanax senticosus, was evaluated for anti-inflammatory and antinociceptive activities. Liriodendrin (5, 10 mg/kg/day, p. o.) significantly inhibited the increase of vascular permeability induced by acetic acid in mice and reduced an acute paw edema induced by carrageenan in rats. When the analgesic activity was measured by the acetic acid-induced writhing test and hot plate test, liriodendrin showed a dose dependent inhibition in animal models. In addition, syringaresinol, the hydrolysate of liriodendrin, more potently inhibited the LPS-induced production of NO, PGE 2 and TNF-alpha production of macrophages than liriodendrin. Consistent with these observations, the expression level of iNOS and COX-2 enzyme was decreased by syringaresinol in a concentration-dependent manner. These results suggest that the anti-inflammatory and antinociceptive effects of liriodendrin after oral administration were attributable to the in vivo transformation to syringaresinol, which may function as the active constituent. PMID- 12898417 TI - Antimicrobial constituents from goldenseal (the Rhizomes of Hydrastis canadensis) against selected oral pathogens. AB - Two new C-methyl flavonoids, 6,8-di- C-methylluteolin 7-methyl ether (1) and 6- C methylluteolin 7-methyl ether (2), were isolated from a commercially available sample of the roots of Hydrastis canadensis, along with seven known compounds, berberine (3), beta-hydrastine (4), canadine (5), canadaline (6), isocorypalmine (7), canadinic acid (8), and beta-sitosterol 3- O-beta- D-glucoside (9). The structures of the new compounds 1 and 2 were elucidated on the basis of their spectral data including 1D and 2D NMR techniques. Of these isolates, berberine (3) and, to a lesser extent, 1 and 2, showed antimicrobial activity when evaluated against the oral pathogens Streptococcus mutans and Fusobacterium nucleatum. Berberine (3) exhibited an additive antimicrobial effect when tested against S. mutans in combination with 1. PMID- 12898418 TI - Inhibitory effect of sterols from Ruprechtia triflora and diterpenes from Calceolaria pinnifolia on the growth of Mycobacterium tuberculosis. AB - Bioactivity-guided fractionation of the CH 2 Cl 2 /MeOH extract of the aerial part of Ruprechtia triflora Griseb. led to the identification of several sterols and a triterpene as the active components against Mycobacterium tuberculosis. This is the first report of a chemical investigation of a member of the genus Ruprechtia. The novel acylated sterol, 5alpha,8alpha-epidioxyergosta-6,22-dien 3beta-yl stearate, was isolated and its structure determined on the basis of spectral evidence including NMR (especially selective 1D NOE, selective 1D TOCSY, HSQC, HMBC) and MS (HR-FAB). In addition, several terpenes obtained from Calceolaria pinnifolia Cav. were also evaluated for their antimycobacterial activity. In a microplate alamar blue assay, sterols from R. triflora were found to be active with MIC values ranging from 2 - 128 microg/mL, with 5alpha,8alpha epidioxyergost-6,22-dien-3beta-ol, 5alpha,8alpha-epidioxystigmasta-6,22-dien 3beta-ol and stigmast-4-en-6beta-ol-3-one being the most active, each with an MIC value of 2 microg/mL. Among the diterpenes from C. pinnifolia, 19 malonyloxydehydroabietinol and 19-methylmalonyloxy- ent-isopimara-8(9),15-diene were most active each with an MIC value of 4 microg/mL. MIC values for the triterpenes 3-epi-ursolic acid and 3-epi-oleanolic acid from C. pinnifolia were determined to be 8 and 16 microg/mL, respectively. PMID- 12898419 TI - The antihyperglycaemic activity of berberine arises from a decrease of glucose absorption. AB - The mechanism of action of berberine as an antihyperglycaemic agent was investigated in the Caco-2 cell line. Berberine was found to effectively inhibit the activity of disaccharidases in Caco-2 cells. It also decreased sucrase activity after preincubation with Caco-2 cells for 72 hours. However gluconeogenesis and glucose consumption of Caco-2 cells were not influenced. 2 Deoxyglucose transporting through Caco-2 cell monolayers was decreased by berberine but the effect was not statistically significant. These results suggest that the antihyperglycaemic activity of berberine is at least partly due to its ability to inhibit alpha-glucosidase and decrease glucose transport through the intestinal epithelium. PMID- 12898420 TI - The sedative effects and mechanism of action of cedrol inhalation with behavioral pharmacological evaluation. AB - It has been reported that cedarwood oil has sedative effects when inhaled. In this study, we evaluated sedative effects of inhaled cedrol, which is a major component of cedarwood oil. Accumulative spontaneous motor activity was significantly decreased in the cedrol-exposed Wistar rats. Similar results were confirmed in caffeine-treated Wistar rats, spontaneously hypertensive rats (SHR), and ddY mice. In addition, exposure to cedrol prolonged pentobarbital-induced sleeping time in Wistar rats. To investigate whether cedrol, which has a very faint aroma, affects the olfactory system, the nasal cavities of Wistar rats were treated with zinc sulfate to reduce olfactory function. Two days later, the pentobarbital-induced sleep time was measured as described above. Compared to intact rats, the sleep prolongation effect was decreased in a lavender-roman chamomile mixed oil exposure positive control group, indicating that olfactory function was impaired. In contrast, prolongation of the sleeping time did not change in the cedrol exposure group. The above findings indicate that cedrol inhalation had marked sedative effects regardless of the animal species or the functional state of the autonomic nerves, suggesting that the mechanism of action is via a pathway other than the olfactory system. PMID- 12898421 TI - Inhibitory effects of rosmarinic acid on Lck SH2 domain binding to a synthetic phosphopeptide. AB - In the course of screening inhibitors from the methanol (MeOH) extracts of 168 medicinal plants against lymphocyte cell-specific kinase (Lck) Src -homology 2 (SH2) binding to a synthetic phosphotyrosine-containing peptide (phosphopeptide), we isolated rosmarinic acid from the MeOH extract of Prunella vulgaris, which showed specific inhibitory activity. The IC 50 value for Lck SH2 binding to phosphopeptide (SGSGEEPQpYEEIPI) of hamster polyomavirus middle-sized tumor (hmT pY324) was 7 microM. However, even at concentrations of 0.1 to 1000 microM, no significant inhibitions were observed against other SH2 domains binding such as the growth factor receptor binding protein 2 (Grb2) SH2 domain to phosphopeptide of Shc and phospholipase Cgamma1 (PLCgamma1) SH2 domain to translational elongation factor 1alpha (EF1alpha) C-terminal. Rosmarinic acid inhibited interleukin-2 (IL-2) gene expression by 50 % at a concentration of 8 microM in Jurkat cells stimulated with anti-CD3 and anti-CD4 antibodies. FK506 and cyclosporin A (CsA) employed as positive controls showed less than 30 % inhibition at the same concentration. In addition, rosmarinic acid inhibited the intracellular [Ca 2+] i increase in Jurkat cells after T cell activation in a dose-dependent manner at concentrations of 1.4 to 140 microM of rosmarinic acid, which is one of the earliest responses of antigen-specific T cell receptor (TCR) and of the upstream pathway of IL-2 expression. Taken together, these results suggest that rosmarinic acid has the potential to specifically inhibit Lck SH2 domain binding to its cognate ligand, including ZAP-70, Cbl, HS-1, and PLCgamma1, and Lck-dependent Ca 2+ signaling pathway of its downstream effector and finally to modulate IL-2 gene expression after T cell activation. PMID- 12898422 TI - Phylogenetic relationship in the genus Panax: inferred from chloroplast trnK gene and nuclear 18S rRNA gene sequences. AB - Chloroplast trnK gene and nuclear 18S rRNA gene sequences of 13 Panax taxa, collected mainly from Sino-Japanese floristic region, were investigated in order to construct phylogenetic relationship and to assist taxonomic delimitation within this genus. The length of trnK gene sequence varied from 2537 bp to 2573 bp according to the taxa, whereas matK gene sequences, embedded in the intron of trnK gene, were of 1512 bp in all taxa. Species-specific trnK/ matK sequence provided much insight into phylogeny and taxonomy of this genus. 18S rRNA gene sequences were of 1808 or 1809 bps in length, only 9 types of 18S rRNA sequences were observed among 13 taxa. Parsimony and neighbor-joining analyses of the combined data sets of trnK-18S rRNA gene sequences yielded a well-resolved phylogeny within genus Panax, where three main clades were indicated. P. pseudoginseng and P. stipuleanatus formed a sister group located at a basal position in the phylogenetic tree, which suggested the relatively primitive position of these two species. Monophyly of P. ginseng, P. japonicus (Japan) and P. quinquefolius, which are distributed in northern parts of Asia or America, was well supported (Northern Clade). The remaining taxa distributed in southern parts of Asia formed a relatively large clade (Southern Clade). The taxonomic debated taxa traditionally treated as subspecies or varieties of P. japonicus or P. pseudoginseng showed various nucleotide sequences, but all fell into one cluster. It might suggest these taxa are differentiated from a common ancestor and are in a period of high variation, which is revealed not only on morphological appearance, but also on molecular divergence. By comparing trnK and 18S rRNA gene sequences among 13 Panax taxa, a set of valuable molecular evidences for identification of Ginseng drugs was obtained. PMID- 12898423 TI - Bioactive coumarins from the leaves of Murraya omphalocarpa. AB - Using antiplatelet aggregation as a guide to fractionation, eight coumarins, omphalocarpinol (1), 5,7-dimethoxy-8-(3'-methyl-2'-oxobutyl)coumarin (2), murralongin, murrayanone, omphamurin (5), murragleinin, mexoticin, and murrangatin, were isolated from the leaves of Murraya omphalocarpa. Compound 1 is new, and 5 is a new enantiomer of omphamurin. The structures of these compounds were elucidated on the basis of spectroscopic techniques, and the structure of compound 1 was confirmed by X-ray crystallographic analysis. Among them, compounds 1, 2 and 5 exhibited significant antiplatelet aggregation activity. PMID- 12898424 TI - Flavonoids and isoflavonoids with antiplasmodial activities from the root bark of Erythrina abyssinica. AB - From the root bark of Erythrina abyssinica a new pterocarpene [3-hydroxy-9 methoxy-10-(3,3-dimethylallyl)pterocarpene] and a new isoflav-3-ene [7,4' dihydroxy-2',5'-dimethoxyisoflav-3-ene] were isolated. In addition, the known compounds erycristagallin, licoagrochalcone A, octacosyl ferulate and triacontyl 4-hydroxycinnamate were identified. The structures were determined on the basis of spectroscopic evidence. The crude extract and the flavonoids and isoflavonoids obtained from the roots of this plant showed antiplasmodial activities. PMID- 12898425 TI - Sesquiterpenes and other constituents from the aerial parts of Inula japonica. AB - Three new sesquiterpenes, 1beta-hydroxy-8beta-acetoxycostic acid methyl ester, 1beta-hydroxy-8beta-acetoxyisocostic acid methyl ester and 1beta-hydroxy 4alpha,11alpha-eudesma-5-en-12,8beta-olide, along with fourteen known compounds were isolated from the aerial parts of Inula japonica. The structures of these new compounds were elucidated by spectroscopic methods (IR, EIMS, HRMS, 1D and 2D NMR). 1,6alpha-dihydroxy-4alphaH-1,10-secoeudesma-5(10),11(13)-dien-12,8beta olide exhibited appreciable cytotoxic activity against cultured SMMC-7721 (human hepatoma cell) and HO-8910 (human ovarian carcinoma cell) with IC 50 values of 52.22 and 21.32 microg/mL, while 5alphaH-eudesma-4(15),11(13)-dien-12,8beta-olide showed remarkable cytotoxic activity against cultured SMMC-7721 and HO-8910 with IC 50 values of 6.21 and 5.28 microg/mL, respectively. PMID- 12898426 TI - A cytotoxic butenolide, two new dolabellane diterpenoids, a chroman and a benzoquinol derivative formosan Casearia membranacea. AB - Investigation of a cytotoxic chloroform-soluble fraction of the stem of Casearia membranacea (Flacourtiaceae) led to the isolation of five new compounds, including one butenolide, casealactone (1), one chroman, caseamemin (2), two dolabellane diterpenoids, casearimene A (3) and casearimene B (4), one benzoquinol ether, casearinone (5), together with fifteen known compounds, including two amides, N- trans-feruloyltyramine (6) and N- cis-feruloyltyramine (7), six steroids, beta-sitosterol (8), stigmast-5-ene-3beta,7alpha-diol (9), stigmast-5-ene-3beta,7beta-diol (10), stigmastane-3beta,5alpha,6beta-triol (11), beta-sitostenone (12), beta-sitosterol 3- O-beta-glucoside (13), two triterpenoids, squalene (14) and friedelin (15), one lignan, (+/-)-syringaresinol (16), two benzenoids, syringaldehyde (17) and vanillic acid (18), one ester, methyl hexadecanoate (19), and anthraquinone (20), respectively. Among these isolates, 1 showed cytotoxicity against P-388 and HT-29 cancer cell lines in vitro, and 6 and 7 showed cytotoxicity against the P-388 cancer cell line. The structures of these compounds were determined by means of spectroscopic techniques, and the structure of 3 was confirmed by X-ray crystallographic analysis. PMID- 12898427 TI - In vitro TGF-beta1 antagonistic activity of ursolic and oleanolic acids isolated from Clerodendranthus spicatus. AB - The mechanism of action of the aerial parts of Clerodendranthus spicatus (Thunb.) C.Y. Wu, [syn. Orthosiphon aristatus (Blume) Miq,] a medicinal plant used in China to treat human renal disease, was investigated. The aqueous and methanol crude extracts exhibited dose-dependent inhibitory activity on 125I-TGF-beta1 binding to its receptor in Balb/c 3T3 cells. Subsequent bioassay-guided fractionation led to identification of two known triterpenoidal constituents, ursolic and oleanolic acids. Ursolic and oleanolic acids inhibited the binding of 125I-TGF-beta1 to its receptor with IC 50 values of 6.9 +/- 0.8 and 21.0 +/- 2.3 microM, respectively. The results suggest that TGF-beta1 antagonistic activity is responsible, at least in part, for the therapeutic efficacy of this plant to treat humans with renal disease. PMID- 12898428 TI - Antibacterial new clerodane diterpenes from the surface of Haplopappus foliosus. AB - A biologically monitored fractionation of the resinous exudate extract of Haplopappus foliosus DC. is reported. Purification of the two active fractions yielded 2-alpha-hydroxy- cis-cleroda-3,13(Z),8(17)-trien-15-oic acid (1) and 2 alpha-acetoxy- cis-cleroda-3,13(Z),8(17)-trien-15-oic acid (2), two new clerodane diterpenes. PMID- 12898429 TI - Aporphine alkaloids from Guatteria spp. with leishmanicidal activity. AB - Fractionation of Guatteria amplifolia yielded the alkaloids xylopine (1), nornuciferine (4), lysicamine (6), and laudanosine (5). Fractionation of Guatteria dumetorum yielded the alkaloids cryptodorine (2) and nornantenine (3). Compounds 1-4 demonstrated significant activity against Leishmania mexicana and L. panamensis. Xylopine (1) was among the most active compounds (LD 50 = 3 microM) and showed a 37-fold higher toxicity towards L. mexicana than macrophages, the regular host cells of Leishmania spp. PMID- 12898430 TI - Arbuscular mycorrhizal inoculation enhances survival rates and growth of Micropropagated plantlets of Echinacea pallida. AB - In an attempt to induce positive effects on the acclimatization of in vitro propagated Echinacea pallida, four arbuscular mycorrhizal (AM) fungi, Glomus mosseae, Gigaspora ramisporophora, Scutellospora fulgida and Entrophospora colombiana were selected to aid the soil adaptation process. Fungal inocula affected the survival of E. pallida plantlets ranging from 83 % to 92 %, depending on the AM species and also contrasting with 58 % survival of the non inoculated plantlets. Growth and development were faster in mycorrhizal treated plantlets than in non-treated ones, especially among those treated with Glomus mossae and Scutellospora fulgida. The presence of well-formed arbuscules and vesicles in Echinacea infested roots was confirmed by microscopic examinations in addition to 90 % success in the survival rate of vigorous plants indicated that mycorrhization is a valuable tool to overcome Echinacea acclimatization shock. PMID- 12898431 TI - Two new ellagic acid rhamnosides from Punica granatum heartwood. AB - From Punica granatum heartwood two new ellagic acid rhamnosides, 3- O methylellagic acid 4-O-alpha-L-rhamnopyranoside (1) and 3,4'-O-dimethylellagic acid 4-O-alpha-L-rhamnopyranoside (2) were isolated together with brevifolincarboxylic acid, 3-O-methylellagic acid and 4,4'-O-dimethylellagic acid. PMID- 12898432 TI - Genetic characterization of the three medicinal Echinacea species using RAPD analysis. AB - The three medicinal species of the Echinacea genus, E. angustifolia DC., E. pallida (Nutt.) Nutt. and E. purpurea (L.) Moench were distinguished using the RAPD (random amplified polymorphic DNA) technique. Species-specific markers were identified from amplicons obtained with four of the twenty 10-mer primers contained in the Operon RAPD kit A. In particular, one marker was identified for E. angustifolia (OPA 20, 1800 pb) and E. pallida (OPA 10, 600 pb) and three markers for E. purpurea (OPA 11 : 1250 pb; OPA 17 : 750, 1800 pb). Genetic distance analysis indicated a high degree of difference among the three species with a relative lower difference between E. angustifolia and E. pallida. PMID- 12898433 TI - Estimating Haemophilus influenzae type b vaccine effectiveness in England and Wales by use of the screening method. AB - In October 1992, Haemophilus influenzae type b (Hib) conjugate vaccine was introduced to infants in the United Kingdom with a "catch-up" program for those aged <4 years. Initially, the rate of invasive Hib disease decreased dramatically but has been increasing since 1999. To determine possible reasons for this increase, the effectiveness of Hib conjugate vaccine was estimated by use of the screening method. Between October 1993 and December 2001, a total of 443 cases of Hib infection occurred in children eligible for vaccination; 363 (82%) were fully vaccinated. Vaccine effectiveness was estimated to be 56.7% (95% confidence interval, 42.5-67.4). Effectiveness was lower in children vaccinated during infancy, compared with those who were vaccinated during the catch-up campaign (P=.0033), declined with time since vaccination (P=.0008), and was lower in children born during 2000-2002, compared with other children scheduled for infant vaccination (P=.0041). Use of a catch-up vaccination program enhanced the control of Hib infection in England and Wales. Since 1999, however, low effectiveness in infants, declining effectiveness with age, and the use of lower-efficacy vaccines have contributed to increased rates of Hib infection. The potential role of boosters needs to be considered. PMID- 12898434 TI - Natural variation in populations of persistently colonizing bacteria affect human host cell phenotype. AB - The highly diverse bacterium Helicobacter pylori, which persistently colonizes the human stomach, provides models to study the role of genome plasticity in host adaptation. Within H. pylori populations from 2 colonized individuals, intragenomic recombination between cagA DNA repeat sequences leads to deletion or duplication of tyrosine phosphorylation sites in the CagA protein, which is injected by a type IV secretion system into host cells. Experimental coculture of gastric epithelial cells with the strains containing these naturally occurring CagA phosphorylation site variants induced markedly divergent host cell morphologic responses. Mutants were constructed in which a phosphorylation site was either added or deleted in the expressed CagA protein; coculture studies confirmed that the naturally occurring differences in CagA phosphorylation are responsible for the observed phenotypic variation. These findings indicate that within an individual host, intragenomic recombination between H. pylori repetitive DNA produces strain variants differing in their signals to host cells. PMID- 12898435 TI - Plasminogen enhances virulence of group A streptococci by streptokinase-dependent and streptokinase-independent mechanisms. AB - Interactions between host plasminogen (Plg) and streptokinase (SK) secreted by group A streptococci (GAS) have been hypothesized to promote bacterial invasion of tissues. The virulence of GAS strain UMAA2616, after being subcutaneously inoculated into mice, was studied. Skin lesions and mortality were observed after inoculation of 7x106 cfu. Coadministration of human Plg with UMAA2616 markedly increased virulence. SK-deficient UMAA2616 (UMAA2616-SK(-)) was generated. Mean skin-lesion area and mortality, after bacterial inoculation (3x105 cfu), were significantly greater with UMAA2616 in the presence of human Plg than with UMAA2616-SK(-) in the presence of human Plg (P=.0001). Human Plg also enhanced UMAA2616-SK(-) virulence. Exogenous human Plg enhanced the virulence of MGAS166, a human clinical isolate. These findings suggest that SK-Plg interactions are an important determinant of GAS invasiveness in vivo and that both SK and host Plg activators appear to promote virulence of GAS by catalyzing plasmin formation. PMID- 12898436 TI - Genetic susceptibility to enteroaggregative Escherichia coli diarrhea: polymorphism in the interleukin-8 promotor region. AB - Enteroaggregative Escherichia coli (EAEC) infection can be identified in 26% of travelers with diarrhea and is associated with fecal interleukin (IL)-8 production. We hypothesized that single-nucleotide polymorphisms (SNPs) in the IL 8 gene are associated with EAEC-related symptoms. Fecal IL-8 production and IL-8 SNPs at 5 loci were identified in 69 US students who remained in Mexico for 5 weeks; 23 subjects had EAEC-associated diarrhea, 7 were asymptomatic EAEC carriers, 22 had nonspecific diarrhea, and 17 were asymptomatic without an enteropathogen. The chances of having EAEC-associated diarrhea were significantly increased among those with the AA genotype at the -251 position (odds ratio [OR], 208.51; 95% confidence interval [CI], 28.5-1525.36) and among those with AT genotype (OR, 14.3; 95% CI, 1.98-105.74), compared with those with the TT genotype at the -251 position. Among subjects with EAEC-associated diarrhea, the AA genotype at the -251 position produced greater concentrations of fecal IL-8 than those with the AT or TT genotype (P=.0053). In the present study, the AA genotype at the -251 position was associated with the occurrence of EAEC associated diarrhea and increased levels of fecal IL-8. PMID- 12898437 TI - Genotypic and phenotypic analysis of type III secretion system in a cohort of Pseudomonas aeruginosa bacteremia isolates: evidence for a possible association between O serotypes and exo genes. AB - The type III secretion system (TTSS) of Pseudomonas aeruginosa was characterized genetically and phenotypically in 92 epidemiologically unrelated bacteremic strains. Four groups of strains (TTSS types) were defined according to the level of type III protein secretion and kinetics of cytotoxicity. Type 1 strains (n=26) were highly and rapidly cytotoxic and secreted ExoU, type 2 strains (n=48) exhibited slower cytotoxic rates and expressed ExoS but not ExoU, type 3 strains (n=14) were poorly cytotoxic, and type 4 strains (n=4) were not cytotoxic. Type 3 and 4 strains did not have detectable secretion phenotype; however, some type 4 strains were able to reach a level of cytotoxicity similar to that of type 1 and type 2 strains when complemented in trans by a functional exsA gene. A statistically significant association (P<.001) was found between TTSS types and detection of the mutually exclusive exoU and exoS genes. In addition, 24 of 25 serotype O:1, O:10, and O:11 strains contained exoU, whereas 54 of 55 serotype O:3, O:4, O:6, O:12, and O:16 strains contained exoS (P<.001). Our results demonstrate correlations among exoU or exoS genotype, TTSS phenotype, and O serotype in bacteremic P. aeruginosa isolates. PMID- 12898438 TI - Detection of Staphylococcus aureus biofilm on tampons and menses components. AB - Culturing has detected vaginal Staphylococcus aureus in 10%-20% of women. Because growth mode can affect virulence expression, this study examined S. aureus biofilm occurrence in 44 paired-tampon and vaginal-wash-specimens from 18 prescreened women, using fluorescent in situ hybridization (FISH). All 44 specimens were also analyzed for S. aureus by standard culturing on mannitol salt agar, which produced positive results for 15 of the 44 specimens. FISH detected S. aureus cells in all 44 specimens, and S. aureus biofilm was observed in 37 of the 44 specimens. Independent confirmation of the presence of S. aureus in specimens from all 18 women was also obtained by amplification, via polymerase chain reaction, of an S. aureus-specific nuclease gene. The results of this study demonstrate that S. aureus biofilm can form on tampons and menses components in vivo. Additionally, the prevalence of vaginal S. aureus carriage may be more prevalent than what is currently demonstrated by standard culturing techniques. PMID- 12898439 TI - A randomized, double-blinded, placebo-controlled trial of intermittent administration of interleukin-2 and prednisone in subjects infected with human immunodeficiency virus. AB - Intermittent administration of interleukin (IL)-2 produces significant and sustained increases in CD4(+) T lymphocyte count in human immunodeficiency virus (HIV)-infected subjects but can be associated with dose-limiting toxicities. The primary objective of this study was to determine whether concomitant administration of prednisone could decrease these toxicities. HIV-seropositive adults receiving highly active antiretroviral therapy (HAART) were randomized to receive either (1) intermittent subcutaneous IL-2 and placebo, (2) intermittent subcutaneous IL-2 and prednisone, (3) intermittent prednisone, or (4) intermittent placebo. Prednisone decreased levels of proinflammatory cytokines during IL-2 cycles but, despite induction of expression of CD25, blunted increases in IL-2-associated CD4(+) T lymphocyte count. Whereas intermittent administration of IL-2 reduced basal proliferation of CD4(+) T cells, this effect was inhibited by prednisone, suggesting that prednisone potentially interferes with IL-2's long-term effects on survival of T lymphocytes. PMID- 12898440 TI - Clinical impact of the M184V mutation on switching to didanosine or maintaining lamivudine treatment in nucleoside reverse-transcriptase inhibitor-experienced patients. AB - Virologic outcome among 104 lamivudine (3TC)-experienced individuals infected with human immunodeficiency virus type 1 who switched to a didanosine (ddI) containing triple- or quadruple-drug regimen was compared with those who continued receiving a 3TC-containing regimen. A significantly increased independent risk of virologic failure was associated with continuing a 3TC containing regimen. In addition, most patients for whom the ddI-containing regimen failed lost the M184V/I mutation. These results show that ddI continues to provide activity against viruses with the M184V/I mutation and suggest that the presence of the M184V/I mutation should not preclude the use of ddI in nucleoside-experienced patients. PMID- 12898441 TI - Antiretroviral concentrations in untimed plasma samples predict therapy outcome in a population with advanced disease. AB - This study was designed to examine the relationship between untimed antiretroviral concentrations measured in plasma samples collected for virus-load testing and response to highly active antiretroviral therapy. Plasma nonnucleoside reverse-transcriptase-inhibitor and protease-inhibitor concentrations were retrospectively measured in all virus-load plasma samples collected during the first year of therapy, for 122 patients in British Columbia, Canada, who initiated therapy between August 1996 and September 1999 and who had CD4 counts <50 cells/micro L. Drug levels were designated a priori as "low" if the concentrations were below the published Ctrough-SD. A single low drug level measured shortly after initiation of therapy (median, 6 weeks) is common (30%) and is predictive of both more-rapid immunological failure (P=.06) and failure to achieve virologic success during the first year of therapy (P=.01). These results may reflect incomplete adherence, since a strong association (P<.001) was found between low drug levels and an imperfect prescription-refill record (<95%). PMID- 12898442 TI - Independent levels of cell-free and cell-associated human immunodeficiency virus 1 in genital-tract secretions of clinically asymptomatic, treatment-naive African women. AB - Using ultrasensitive polymerase chain reaction-based techniques, we assessed levels of cell-free and cell-associated human immunodeficiency virus (HIV)-1 in paired blood and genital samples of 30 clinically asymptomatic, treatment-naive women. Levels of HIV-1 RNA in cervicovaginal-lavage samples were positively correlated with those in plasma samples (r=.50; P=.008), whereas levels of HIV-1 DNA in genital samples were loosely correlated with those in blood samples (r=.31; P=.041). In plasma of peripheral blood, levels of HIV-1 DNA were positively correlated with those of HIV-1 RNA (r=.64; P<.001), whereas no correlation between HIV-1 DNA and HIV-1 RNA was evident in genital secretions. Our results indicate that levels of HIV-1 RNA and HIV-1 DNA are unrelated in the genital tracts of treatment-naive women and suggest that the level of genital HIV 1 RNA is influenced by systemic viral replication-in contrast to genital HIV-1 provirus, which may be influenced as well by local cofactors triggering the migration of HIV-infected cells originating from the cervicovaginal submucosa. These features may be relevant for an understanding of HIV-1 transmission in heterosexual individuals. PMID- 12898444 TI - Pathogenesis of enterovirus 71 brainstem encephalitis in pediatric patients: roles of cytokines and cellular immune activation in patients with pulmonary edema. AB - Taiwan experienced several epidemics of enterovirus 71 (EV71) infections, which were associated with brainstem encephalitis (BE) and pulmonary edema (PE). To elucidate the role of immune mechanisms in the pathogenesis of BE caused by EV71 and its fatal complication, PE, we analyzed the laboratory findings, cytokine, and immunophenotypes of 73 EV71-infected patients with BE. Patients were stratified by disease: PE (n=14), autonomic nervous system (ANS) dysregulation (n=25), and isolated BE (n=34). The mortality rate for PE was 64.3%. Leukocytosis and thrombocytosis were significantly more frequent among patients with PE. A significant elevation of plasma interleukin (IL)-10, IL-13, and interferon (IFN) gamma levels observed in patients with PE. Patients with PE also had lower circulating CD4(+) T cells, CD8(+) T cells, and natural killer (NK) cells. An extensive peripheral and central nervous system inflammatory response with abnormal IL-10, IL-13, and IFN-gamma cytokine production and lymphocyte depletion appears to be responsible for the pathogenesis of EV71-associated PE. PMID- 12898443 TI - Increased risk of high-grade cervical squamous intraepithelial lesions and invasive cervical cancer among African women with human immunodeficiency virus type 1 and 2 infections. AB - To assess the risk of prevalent high-grade cervical squamous intraepithelial lesions (HSILs) or invasive cervical cancer (ICC) associated with human immunodeficiency virus (HIV) type 1, HIV-2, and human papillomavirus (HPV) infections, HIV load, and CD4 cell count, we studied 4119 women attending an outpatient clinic in Senegal. HIV infection was associated with increased rates of cervical infection with high-risk HPVs. Among women infected with high-risk HPVs, those with HIV-1 (odds ratio [OR], 2.2; 95% confidence interval [CI], 1.0 4.8), HIV-2 (OR, 6.0; 95% CI, 2.1-17.1), or dual HIV infection (OR, 8.0; 95% CI, 2.0-31.5) were more likely to have HSILs or ICC diagnosed than were HIV-negative women; this association was not observed among women not infected with high-risk HPVs. Among women with HIV, higher HIV plasma RNA loads and lower CD4 cell counts were associated with high-risk HPV infection and degree of cervical abnormality. Furthermore, HIV-2-positive women were more likely to have HSILs (OR, 3.3; 95% CI, 0.9-12.4) or ICC (OR, 7.9; 95% CI, 1.1-57) than were HIV-1-positive women. PMID- 12898445 TI - Prevalence of chronic hepatitis B and incidence of acute hepatitis B infection in human immunodeficiency virus-infected subjects. AB - We determined incidence and risk factors for acute and chronic hepatitis B virus (HBV) infection and HBV vaccination rates among human immunodeficiency virus (HIV)-infected subjects from the Adult/Adolescent Spectrum of HIV Disease Project, during 1998-2001. Among 16,248 HIV-infected patients receiving care, the incidence of acute HBV was 12.2 cases/1000 person-years (316 cases), was higher among black subjects (rate ratio [RR], 1.4; 95% confidence interval [CI], 1.0 2.0), subjects with alcoholism (RR, 1.7; 95% CI, 1.2-2.3), subjects who had recently injected drugs (RR, 1.6; 95% CI, 1.1-2.4), and subjects with a history of AIDS-defining conditions (RR, 1.5; 95% CI, 1.2-1.9) and was lower in those taking either antiretroviral therapy (ART) with lamivudine (RR, 0.5; 95% CI, 0.4 0.6), ART without lamivudine (RR, 0.5; 95% CI, 0.3-0.7), or >/=1 dose of HBV vaccine (14% of subjects) (RR, 0.6; 95% CI, 0.4-0.9). Prevalence of chronic HBV was 7.6% among unvaccinated subjects. HBV rates in this population were much higher than those in the general population, and vaccination levels were low. HBV remains an important cause of comorbidity in HIV-infected persons, but ART and vaccination are associated with decreased disease. PMID- 12898446 TI - Quantification of the influenza virus load by real-time polymerase chain reaction in nasopharyngeal swabs of patients treated with oseltamivir. AB - Influenza A virus load was assessed by real-time polymerase chain reaction (PCR) in nasopharyngeal swabs of infected patients treated with oseltamivir. The mean pretreatment virus load was significantly lower in the 24 patients (group A) who initiated treatment within 24 h of the onset of symptoms than it was in the 26 patients (group B) who initiated treatment between 24 and 48 h (1.6x105 vs. 8.4x105 copies/600 ng of total RNA, P=.04); after 48 h of treatment, twice as many patients in group B still had a positive PCR result, compared with patients in group A (42.3% vs. 20.7%). These virological results support the clinical benefit provided by early therapeutic intervention of influenza illness. PMID- 12898447 TI - Cytomegalovirus (CMV) glycoprotein B genotypes and response to antiviral therapy, in solid-organ-transplant recipients with CMV disease. AB - Cytomegalovirus (CMV) can be classified into 4 glycoprotein B (gB) genotypes, on the basis of sequence variation in the UL55 gene. We assessed the effect that CMV gB genotype has on virologic and clinical response to therapy, in 50 solid-organ transplant recipients with CMV disease. CMV loads were determined at regular intervals after the start of therapy. Genotype results were correlated with CMV load kinetics in response to therapy with ganciclovir. At the onset of treatment, the distribution of CMV gB genotypes was as follows: gB1, 19/50 (38%); gB2, 9/50 (18%); gB3, 12/50 (24%); gB4, 2/50 (4%); and mixed-genotype infection, 8/50 (16%). Between viral genotype groups, time to clearance of CMV, failure to clear CMV, and calculated CMV-load half-life after the start of therapy were not significantly different. The CMV gB genotype did not affect the rate of disease recurrence or occurrence of tissue-invasive disease. It appears that the gB genotype, which causes CMV disease, does not significantly influence CMV-load kinetics or clinical response to therapy. PMID- 12898448 TI - Selective effects of interleukin (IL)-15 on antifungal activity and IL-8 release by polymorphonuclear leukocytes in response to hyphae of Aspergillus species. AB - The effects of interleukin (IL)-15 on human polymorphonuclear leukocyte (PMNL) activity against Aspergillus fumigatus and Aspergillus flavus were investigated. Pretreatment with IL-15 for 2 h increased PMNL oxidative burst, as measured by superoxide anion (O(2)(-)) release in response to A. fumigatus (P<.05) but not to A. flavus. However, after 22-h, but not 2-h, treatment with IL-15, there was significant enhancement in PMNL-mediated hyphal damage to A. fumigatus. Furthermore, 22-h exposure to IL-15 mediated an enhanced release of IL-8 from PMNLs challenged with hyphae of A. fumigatus and A. flavus (P<.05). In contrast, IL-15 treatment did not affect the release of tumor necrosis factor-alpha from PMNLs. The selective time- and species-dependent enhancement of O(2)(-) production and hyphal damage, as well as its induction of IL-8 release, suggest that IL-15 may play an important role in the immunomodulation of host response to invasive aspergillosis. PMID- 12898449 TI - Hydrolytic gene expression during oroesophageal and gastric candidiasis in immunocompetent and immunodeficient gnotobiotic mice. AB - To investigate whether host immunocompetence influences hydrolytic gene expression, we compared secretory aspartyl proteinase gene (SAP) and phospholipase B gene (PLB) expression during gastric candidiasis in immunocompetent and defined immunodeficient gnotobiotic mice, by reverse transcription polymerase chain reaction. The use of immunodeficient gnotobiotic mice with combined defects in T cells and natural killer cells enabled a comprehensive study of virulence gene expression in various mucosal sites during lethal oroesophageal (tongue, palate, and esophagus) and gastric candidiasis. All 10 SAP and both PLB genes were expressed in both immunocompetent and specific immunodeficient mice, which suggests that the absence of important components of the host defense did not alter gene expression during gastric candidiasis. Although similar patterns of gene expression were evident in different oral tissues, we detected specific differences between Candida albicans-infected oroesophageal and gastric tissues and differences at various time points during the progression of gastric candidiasis. PMID- 12898450 TI - Novel mechanism of drug resistance in kala azar field isolates. AB - Clinical resistance to pentavalent antimonial drugs in the form of sodium antimony gluconate (SAG) has become a major problem in the treatment of kala azar (visceral leishmaniasis) in India. The mechanism of resistance is unclear in these clinical isolates, although work has been conducted with Leishmania species mutants selected in vitro by stepwise increase of drug concentration, using antimony-related metal arsenic and, more recently, SAG. In the present study, we investigated the molecular aspect of drug resistance in clinically confirmed SAG resistant field isolates. Our results show that the mechanisms of resistance postulated for laboratory mutants of Leishmania species are not operating in field isolates of Leishmania donovani. Instead, we identified a novel gene amplified in these drug-resistant parasites whose locus is on chromosome 9. The significant finding was that this isolated fragment confers antimony resistance to wild-type Leishmania species after transfection. We speculate that protein phosphorylation may play a role in signal transduction pathway in the parasite after exposure to drug-conferring resistance. PMID- 12898451 TI - Infection of placental trophoblasts by Toxoplasma gondii. AB - How the intracellular parasite Toxoplasma gondii causes placental inflammation and infects the fetus is unknown. By use of a culture model of primary human trophoblasts, we examined the consequences of infection by a virulent strain of T. gondii. Infection fractions (parasitophorous vacuoles per trophoblast nuclei) < or =0.9 were observed 1 day after challenge at an inoculum ratio of T. gondii to nuclei of 10. The culture content of infectious T. gondii increased 45-fold in 48 h. Two days after infection, almost 30% of trophoblast nuclei became apoptotic, and 30%-35% of nuclei were lost. Almost 90% of apoptotic nuclei were not adjacent to a parasitophorous vacuole, suggesting infection protected against apoptosis. However, there was no T. gondii-dependent accumulation of putative cytotoxic factors, such as tumor necrosis factor-alpha, that could mediate paracrine killing. Both mature and immature trophoblasts can be productively infected, and uninfected, but not infected, cells undergo apoptosis. PMID- 12898452 TI - Fc receptor-mediated antibody regulation of T cell immunity against intracellular pathogens. AB - Immunity to intracellular microbial pathogens, including Chlamydia species, is controlled primarily by cell-mediated effector mechanisms, yet, the absence of antibodies results in inefficient microbial clearance. We investigated the hypothesis that certain Fc receptor functions promote the rapid induction of elevated T helper type 1 (Th1) response, which effectively clears chlamydiae. FcR(-/-) mice exhibited a delayed and reduced frequency of Chlamydia-specific Th1 cells, compared to FcR(+/+) mice. In vitro, antichlamydial antibodies increased the rate of Th1 activation by FcR(+/+) but not FcR(-/-) antigen-presenting cells. FcR(-/-) dendritic cells and the T cell-associated IgG2A and IgA mediate enhanced Th1 activation by antibodies. Immunization with chlamydia-antibody complexes induced elevated and protective Th1 response. These results provide a mechanistic basis for requiring both T cell and humoral immune responses in protective immunity and vaccine evaluation. Findings offer a paradigm in host defense wherein different effector components function indirectly to maximize the principal effector mechanism. PMID- 12898456 TI - Alanine administration does not stimulate gluconeogenesis in preterm infants. AB - Gluconeogenesis partially depends on sufficient precursor supply, and plasma alanine concentrations are generally low in preterm infants. Stimulation of gluconeogenesis may contribute to the prevention of hypoglycemia, an important clinical problem in these infants. In this study we evaluated the effect of extra precursor supply on gluconeogenesis in preterm infants. In 11 infants, gestational age < or = 32 weeks, glucose production rate (GPR) and gluconeogenesis were measured using the [6,6-(2)H(2)]glucose dilution technique and mass isotopomer distribution analysis with [2-(13)C]glycerol, respectively. Unlabeled glucose was administered throughout the study period at a rate of 22 micromol. kg(-1). min(-1). Five infants received alanine (1.5 mg. kg(-1). min( 1)) during the last 3 hours of the study protocol, and 6 infants served as controls. In the control group the rate of gluconeogenesis and GPR remained constant at 4.0 +/- 0.3 micromol. kg(-1). min(-1) and 8.3 +/- 0.6 micromol. kg( 1). min(-1), respectively. In the alanine group plasma alanine concentrations increased from 45 +/- 23 to 829 +/- 115 micromol/L (P =.001); gluconeogenesis and GPR did not differ from control: 3.8 +/- 0.2 micromol. kg(-1). min(-1) and 6.4 +/ 2.0 micromol. kg(-1). min(-1), respectively. We conclude that administration of the gluconeogenic precursor alanine does not stimulate gluconeogenesis in preterm infants, despite a sharp increase in plasma alanine concentrations. We speculate either a restricted capacity of the enzymes involved in the gluconeogenic pathway or a low secretion rate of glucoregulatory hormones as causative mechanisms involved in the gluconeogenic pathway in the preterm neonate. PMID- 12898457 TI - The effect of physical exercise on reverse cholesterol transport. AB - High-density lipoproteins (HDL) are recognized for their role in coronary artery disease (CAD) risk reduction. Plasma HDL plays a pivotal role in the reverse cholesterol transport (RCT) process. Physical exercise is well recognized as a modality that affects HDL metabolism. The purpose of this discussion is to describe the effects of physical exercise on RCT. PMID- 12898458 TI - Fat depot-specific expression of adiponectin is impaired in Zucker fatty rats. AB - Adiposity, particularly increased intra-abdominal fat, is a predisposing factor for the development of insulin resistance in obesity and type 2 diabetes. Visceral fat seems to differ from subcutaneous adipose tissue in adipocytokine production. This fat depot-related difference has been viewed as an important mechanism by which adipose tissue exerts its paracrine/autocrine effects on peripheral tissue in modulating insulin sensitivity. We have studied the relative expression of adiponectin in visceral versus subcutaneous fat in Zucker fatty versus lean rats. Visceral fat, as opposed to subcutaneous fat, exhibited relatively higher levels of adiponectin production in lean animals. However, in Zucker fatty rats, adiponectin expression in visceral fat was suppressed to basal levels, which correlated with significantly reduced plasma adiponectin concentrations and increased insulin resistance. These results suggest that an impaired depot-specific expression of adiponectin is a contributing factor for the development of insulin resistance in Zucker fatty rats. PMID- 12898459 TI - Synergistic effects of testosterone and growth hormone on protein metabolism and body composition in prepubertal boys. AB - During human puberty there is a substantial increase in growth hormone (GH) and sex steroidal hormone concentrations, as well as in GH production rates and insulin-like growth factor-I (IGF-I) These studies were designed to investigate some of the interactions of testosterone (T) and GH in the metabolic changes of puberty. Ten boys with severe GH deficiency (GHD) were studied (mean age, 12.5 +/ 0.5 years) using stable isotope infusions, indirect calorimetry, and body composition analysis. After the baseline study, they received 2 doses of T enanthate (50 to 75 mg, intramuscular [IM]), and they were studied again 4 weeks later. The boys were then begun on daily subcutaneous (SC) GH (0.3 mg/kg/wk), while T therapy was continued for another 4 weeks and the studies repeated a third time. The treatment order was randomized. Protein oxidation rates decreased after T alone (-28%, P <.01), decreasing further after combined T/GH treatment ( 36% v baseline, P <.01). The nonoxidative leucine disposal (NOLD), a measure of whole body protein synthesis, increased significantly after combined T/GH regardless of treatment order. The combination of T/GH also resulted in greater changes in body composition than T alone, with comparable decreases in %FM and corresponding increases in fat free mass (FFM). Measures of carbohydrate (CHO) metabolism, including glucose production and oxidation rates, were unaffected by either T or T/GH combination. Plasma IGF-I concentrations increased after T treatment and even more after T/GH combination, regardless of the treatment order. In conclusion GH and T are synergistic on whole body protein anabolism and body composition in males, even at a young age, but the positive effects of T on protein anabolism and body composition appear to need a basal amount of GH for those effects to be observed. GH and T both potentiate the development of the full body composition and metabolic changes of puberty. PMID- 12898460 TI - The effects of dietary restriction on humeral and mandibular bone in SENCAR, C57BL/6, and DBA/2 mice. AB - Dietary restriction (DR) increases the life span and retards the development of age-related disorders. However, the low body mass that accompanies DR is associated with risk factors for fracture that may outweigh the beneficial effects of DR on cellular aging that are mediated, in part, by limiting free radical generation and oxidative damage. We tested the effects of DR in murine models that differ in free radical generation capacity (SENCAR > C57 > DBA). Male mice of each strain were killed at 10 weeks of age (t(0); time zero) or randomized to an ad libitum-fed (AL-fed) or 30% DR feeding regimen for 6 months. The food consumption of AL-fed mice was measured daily. DR mice received 70% of the amount of food consumed by their respective AL-fed mice the previous day. The DR diet was normalized with respect to calcium (Ca), phosphorus (P), and micronutrients. Lean body mass (LBM), bone mineral density (BMD), and bone mineral content (BMC) in the humerus and mandible were determined by PIXImus densitometry. The length and midshaft width of the humerus were determined by direct measurement. There were highly strain- and diet/time-dependent effects on LBM, humerus length, mandibular and humeral BMD, and humeral BMC. The interaction between diet/time and strain was more significant in the humerus than the mandible. All 30% DR mice had lower humeral BMDs and BMCs than their respective AL-fed controls. However, 30% DR C57 and DBA (but not SENCAR) mice had higher humeral BMD and BMC than their respective t(0) controls. There was a linear relationship between LBM and humeral BMD and BMC in both AL-fed and 30% DR mice, suggesting that the lower BMD and BMC in 30% DR mice, relative to AL-fed controls, reflects a physiologic adaptation to lower biomechanical loading. Mandibular BMC in 30% DR C57 (but not DBA or SENCAR) mice was lower than that observed in their AL-fed controls. Mandibular BMD and BMC increased versus t(0) values in 30% DR mice of all strains. PMID- 12898461 TI - Effect of troglitazone on vascular and glucose metabolic actions of insulin in high-sucrose-fed rats. AB - In rats, diets high in simple sugar induce insulin resistance and alter vascular reactivity. The present study was designed to evaluate the effects of 5 weeks treatment with troglitazone on insulin sensitivity, regional hemodynamics, and vascular responses to insulin in chow-fed and high-sucrose-fed rats. Male rats were randomly divided in 4 groups to receive a regular chow diet in the absence (group 1) or presence of troglitazone (0.2% in food; group 2), or a sucrose enriched diet in the absence (group 3) or presence of troglitazone (group 4) for 5 weeks. The rats were instrumented with Doppler flow probes and intravascular catheters to determine blood pressure, heart rate, and regional blood flows. Insulin sensitivity was assessed by the euglycemic hyperinsulinemic clamp technique. Glucose transport activity was examined in isolated muscles. Sucrose feeding was found to induce insulin resistance and to impair the insulin-mediated skeletal muscle vasodilation. Treatment with troglitazone was found to increase whole-body insulin sensitivity in sucrose- and chow-fed rats, but had no effect on skeletal muscle glucose transport activity measured in isolated muscles from both dietary groups. Changes in regional hemodynamics were observed in both dietary cohorts treated with troglitazone, and the hindquarter vasoconstrictor response to insulin noted in sucrose-fed rats was abolished by the treatment. The vascular effects of troglitazone, and its insulin-related attenuating effects on contractile tone, could have contributed, in part, to improve insulin action on peripheral glucose disposal, presumably by improving blood flow distribution and glucose delivery. PMID- 12898462 TI - Apoptosis of human abdominal preadipocytes before and after differentiation into adipocytes in culture. AB - Differentiation of murine 3T3-L1 preadipocytes into adipocytes is associated with the acquisition of apoptotic resistance accompanied by the upregulation of cell survival genes. We have now examined the effect of adipogenesis on apoptotic susceptibility of human abdominal preadipocytes in primary culture. To induce apoptosis, human preadipocytes, or their differentiated counterparts, were serum deprived for 24 or 48 hours. When indicated, ceramide was also used as an apoptotic trigger. Cell death was assessed by enumeration of adherent viable cells, and its apoptotic nature was verified by Hoechst staining and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL). After 48 hours of serum withdrawal, cell death was 26% +/- 4% in preadipocytes and was increased to 41% +/- 4% in differentiated adipocytes (mean +/- SE; n = 7 patients; P <.002). Under serum-free conditions for 24 hours, ceramide-induced cell death was 40% +/- 6% in preadipocytes and increased to 68% +/- 8% in adipocytes (mean +/- SE; P <.01; n = 8 patients). Neuronal apoptosis inhibitor protein (NAIP), an antiapoptotic protein cell survival that increases upon 3T3-L1 adipogenesis, was reduced in human preadipocytes undergoing differentiation (n = 6 patients). Preadipocytes derived from omental versus subcutaneous abdominal fat were more susceptible to apoptosis induced by serum deprivation, 16% +/- 4% versus 31% +/- 3% cell death, respectively (mean +/- SE; P <.02; n = 7 patients). Although the murine 3T3-L1 preadipocyte cell line is a useful model that approximates primary preadipocyte cell biology, our data derived from human preadipocyte studies suggest important differences with respect to the regulation of apoptosis. PMID- 12898463 TI - Export of metabolites from pancreatic islet mitochondria as a means to study anaplerosis in insulin secretion. AB - Recent evidence suggests that the mitochondrial synthesis (anaplerosis) of alpha ketoglutarate or intermediates that can be converted to alpha-ketoglutarate, such as citrate or glutamate, are important for insulin secretion stimulated by metabolizable secretagogues. In order to focus on the productive role of mitochondria (anaplerosis) separate from the consumptive role of the cytosol (cataplerosis) in insulin secretion, pyruvate and other metabolites of insulin secretagogues were added to microgram amounts of mitochondria obtainable from rat or mouse pancreatic islets and the export of metabolites was surveyed. Cellular levels of metabolites in rat islets were also measured. The export of malate from mitochondria was the most responsive to various substrates. The export of citrate did not increase in the presence of pyruvate alone or pyruvate plus glutamate, but malate plus pyruvate caused citrate to be exported. Citrate levels in intact cells did not change with glucose. Glutamate levels did not increase in intact islets in the presence of glucose, thus not providing evidence for glutamate acting as a messenger in glucose-induced insulin secretion. The citrate level may not need to increase in order to provide increased malonyl-coenzyme A for signaling insulin secretion. Unlike many cells, insulin cells probably obtain cytosolic NADPH equivalents by exporting them from mitochondria to the cytosol via a pyruvate malate shuttle or an isocitrate shuttle. The current results suggest that the reason for anaplerosis in insulin secretion is quite complex and not fully explained by current knowledge. PMID- 12898464 TI - Relationship between circulating E-selectin, DD genotype of angiotensin converting-enzyme, and cardiovascular damage in central obese subjects. AB - Fifty-six young central obese patients were investigated to evaluate relationships between soluble E-selectin (sE-S), angiotensin-converting enzyme (ACE) gene polymorphism, left ventricular function and structure, and carotid morphology by determination of sE-S and ACE genotypes. Our results indicated that central obese subjects with concomitant higher levels of sE-S and ACE DD genotype may be characterized by early cardiovascular alterations and then considered a particular subset of subjects at higher risk of cardiovascular disease. PMID- 12898465 TI - Effect of malnutrition during the first year of life on adult plasma insulin and glucose tolerance. AB - There is evidence linking intrauterine growth retardation with increased cardiovascular risk and diabetes mellitus (DM) later in life. However, little is known about the association between malnutrition during the first year of life and metabolic abnormalities in adulthood. The objective of this study was to assess the effect of documented malnutrition during the first year of life on glucose tolerance, plasma insulin, lipid profile, and blood pressure in early adulthood, as well as to assess the interaction between body mass index (BMI) and malnutrition on these variables. A study group of young men with a documented history of malnutrition during their first year of life was recruited from 4 pediatric hospitals in Mexico City and compared with a control group. Subjects included were 52 men, aged 20.2 +/- 3.6 years, with a mean birth weight of 3.0 +/ 0.7 kg and documented malnutrition in their first year of life; controls were 50 men, aged 23.3 +/- 1.8 years, with a mean birth weight of 3.2 +/- 0.5 kg. Insulin and glucose concentrations, fasting and in response to an oral glucose load, plasma lipids, blood pressure, and an insulin sensitivity index (ISI) were measured. The areas under the curves of glucose (AUCG) and insulin (AUCI) were significantly higher in cases (P =.012 and <.002, respectively), independent of birth weight, BMI, or age. BMI was significantly associated with fasting plasma insulin (FPI), AUCI, ISI, triglyceride, and high-density lipoprotein (HDL) cholesterol concentrations in cases, but not in controls. These data suggest that early malnutrition in extrauterine life, independently of birth weight, has an adverse effect on insulin metabolism and glucose tolerance in young men, and it worsens as body mass increases even within the normal range of BMI. Therefore, it is advisable to prevent obesity in individuals exposed to early malnutrition. PMID- 12898466 TI - A novel peroxisome proliferator-activated gamma (PPAR gamma) agonist, CLX-0921, has potent antihyperglycemic activity with low adipogenic potential. AB - Agonists of the nuclear receptor peroxisome proliferator-activated receptor gamma (PPAR gamma) are pharmacologically active antihyperglycemic agents that act by increasing peripheral tissue sensitivity to insulin. Many of these agonists have antihyperglycemic activity that is directly proportional to their ability to bind and activate PPAR gamma; however, recent data bring this relationship into question. In this report we describe a new PPAR gamma agonist, CLX-0921, that is derived from a natural product. This thiazolidinedione (TZD) has a spectrum of activity that differs from commercially available TZDs. It is a weak activator of PPAR gamma (EC(50) of 0.284 micromol/L) compared to rosiglitazone (EC(50) 0.009 micromol/L). Despite this difference, the drug maintains potent glucose uptake activity in vitro and glucose-lowering activity in vivo that is equipotent to that of rosiglitazone. Moreover, CLX-0921 showed a 10-fold reduction in in vitro adipogenic potential compared to rosiglitazone. CLX-0921 also increases glycogen synthesis, an activity not typically associated with rosiglitazone or pioglitazone. Thus CLX-0921 appears to have a distinct spectrum of activity relative to other TZDs. PMID- 12898467 TI - Physiological changes in circulating mannose levels in normal, glucose intolerant, and diabetic subjects. AB - Mannose is an essential hexose that is required for glycoprotein synthesis. Although circulating mannose levels are known to be influenced by metabolic disorders, how physiological levels of mannose fluctuate in normal and diabetic subjects is largely unknown. We describe a new accurate and sensitive assay for determining circulating mannose levels, which we used to measure plasma mannose levels in 273 normal and diabetic (DM) subjects. Our results revealed a clear correlation (r = 0.754) between fasting plasma mannose (FPM) and fasting plasma glucose (FPG) levels. Our mannose assay showed sensitivity and specificity comparable to that seen for hemoglobin A(1c) (HbA(1c)) assay in subjects with impaired glucose tolerance (IGT) or DM whose FPG levels were normal. Mannose levels were found to increase less than glucose levels in response to an oral glucose tolerance test (OGTT). Furthermore, plasma mannose levels did not significantly change following a meal and more closely correlated with the coefficient of variation (CV) of daily glucose levels than did glucose itself. In conclusion, the close correlation between FPM and FPG levels taken together with the small fluctuations seen in plasma mannose in response to glucose suggests that the measurement of mannose using our assay could potentially play a supplementary role in the diagnosis and screening of patients with mild DM. PMID- 12898468 TI - Physical exercise enhances protein kinase C delta activity and insulin receptor tyrosine phosphorylation in diabetes-prone psammomys obesus. AB - We recently reported that physical exercise prevents the progression of type 2 diabetes mellitus in Psammomys obesus, an animal model of nutritionally induced type 2 diabetes mellitus. In the present study we characterized the effect of physical exercise on protein kinase C delta (PKC delta) activity, as a mediator of the insulin-signaling cascade in vivo. Three groups of Psammomys obesus were exposed to a 4-week protocol: high-energy diet (HE/C), high-energy diet and exercise (HE/EX), or low-energy diet (LE/C). None of the animals in the HE/EX group became diabetic, whereas all the animals in the HE/C group became diabetic. After overnight fast, intraperitoneal (IP) insulin (1U) caused a greater reduction in blood glucose levels in the HE/EX and LE/C groups compared to the HE/C group. Tyrosine phosphorylation of insulin receptor (IR), insulin receptor substrate-1 (IRS-1), and phosphatidylinositol 3 kinase (PI3 kinase) was significantly higher in the HE/EX and LE/C groups compared with the HE/C group. Finally, IR-associated PKC delta was higher in the HE/EX and LE/C groups compared to the HE/C group. Coprecipitation of PKC delta with IR was higher in the HE/EX and LE/C groups compared to the HE/C group. Thus, we suggest that 4 weeks of physical exercise results in improved insulin-signaling response in Psammomys obesus accompanied by a direct connection between PKC delta and IR. We conclude that this mechanism may be involved in the preventive effect of exercise on type 2 diabetes mellitus in Psammomys obesus. PMID- 12898469 TI - Incremental area under response curve more accurately describes the triglyceride response to an oral fat load in both healthy and type 2 diabetic subjects. AB - Elevation of postprandial triacylglycerol (TG)-rich plasma lipoproteins is considered potentially atherogenic. Type 2 diabetic patients have exaggerated postprandial TG compared with healthy subjects. Postprandial TG responses to oral fat loads are usually studied as the area under the TG curve. No consensus exists regarding the method of choice when calculating the TG response area. We evaluated the correlation between fasting TG and postprandial TG responses calculated by the trapezoid rule as total area under the curve (AUC) and incremental area under the curve (iAUC). Furthermore, we compared the AUC and iAUC to a 3-point calculation method. Ten healthy subjects and 47 type 2 diabetic patients ingested test meals consisting of an energy-free soup plus 80 g fat and 50 g carbohydrate. TG responses were measured in total plasma, in a chylomicron (CM)-rich fraction and in a CM-poor fraction. In healthy subjects the AUC, but not iAUC, correlated positively to fasting TG. In type 2 diabetic patients a strong correlation was found between fasting TG and AUC, whereas weak associations were found to the iAUCs. The iAUC was strongly correlated to the postprandial TG rise in both groups. The 3-point areas differed significantly from the trapezoid measurements in both healthy and type 2 diabetic subjects. In conclusion, in both healthy and type 2 diabetic subjects total AUC is highly correlated to fasting TG, whereas iAUC more accurately describes the TG response to an oral fat load. The 3-point test seems less suitable for the determination of postprandial response in both healthy and type 2 diabetic subjects. PMID- 12898470 TI - Postprandial lipidemia is normal in non-obese type 2 diabetic patients with relatively preserved insulin secretion. AB - To assess postprandial lipidemia in normotriglyceridaemic type 2 diabetic patients treated with diet only, 12 non-obese patients (8 males, hemoglobin A(1c) [HbA(1c)] 6.80 +/- 0.67%) and 14 controls of similar age, body mass index (BMI), and fasting triglyceride (Tg) were given a test meal (58 g fat, 100,000 IU vitamin A). Fasting low-density lipoprotein (LDL) cholesterol (LDLc), high density lipoprotein (HDL) cholesterol (HDLc), free fatty acids, and apolipoprotein B (apoB), and fasting and postprandial Tg, retinylpalmitate (RP), LDL size, glucose, and insulin were measured. The homeostasis assessment model (HOMA) index and lipoprotein (Lpl) and hepatic (HL) lipase activities were estimated. Patients showed lower fasting HDLc (1.12 +/- 0.26 v 1.40 +/- 0.28 mmol/L, P =.02) and a trend towards smaller LDL particles, which was significant 4 hours postprandially (25.86 +/- 0.40 v 26.16 +/- 0.30 nm, P =.04). The area under the curve of Tg (AUC-Tg) and RP, and Lpl were similar, but HL was higher in patients (156.63 +/- 23.89 v 118 +/- 43.27 U/L, P =.011). HL correlated inversely with LDL size and directly with the HOMA index. In conclusion, normotriglyceridemic type 2 diabetic patients with insulin resistance but relatively preserved insulin secretion show low fasting HDLc and increased HL, but normal postprandial lipidemia. PMID- 12898471 TI - Fasting and daylong triglycerides in obesity with and without type 2 diabetes. AB - Postprandial hypertriglyceridemia associated with insulin resistance is one of the cardiovascular risk factors in obesity and type 2 diabetes. It is not known whether diabetics have a more pronounced postprandial hypertriglyceridemia than obese subjects. Daylong triglyceridemia, representing postprandial lipemia, was determined in obese subjects with and without type 2 diabetes and in lean subjects. Nineteen type 2 diabetics (F/M: 7/12, body mass index [BMI]: 30.6 +/- 5.4 kg/m(2)), 45 obese nondiabetics (F/M: 16/29, BMI: 29.5 +/- 2.6 kg/m(2)) and 78 lean subjects (F/M: 28/50, BMI: 23.7 +/- 2.2 kg/m(2)) measured capillary triglycerides (TGc) during 3 days on 6 fixed time-points each day in an out-of hospital situation. Daylong TGc profiles were calculated as mean integrated area under the TGc-curve (TGc-AUC). Fasting plasma TG were higher in diabetics and obese nondiabetics (1.81 +/- 0.79 and 1.77 +/- 0.80 mmol/L) compared with lean subjects (1.23 +/- 0.67 mmol/L, P <.001). TGc-AUC was similarly increased in both diabetics and obese nondiabetics (35.0 +/- 12.1 and 35.2 +/- 10.6 mmol.1 h/L) compared with lean controls (25.5 +/- 12.0 mmol.1 h/L, P <.001). Self-reported energy intake was not significantly different between the groups. Fasting TGc (r =.87, P <.001) and waist circumference (r =.51, P <.001) were the parameters best associated with TGc-AUC. Using stepwise multiple regression analysis, fasting TGc, BMI, total cholesterol, and high-density lipoprotein (HDL) cholesterol were the best predictors of TGc-AUC, explaining 77% of the variation. The cut-off level for "normal" TGc-AUC, calculated as the 75th percentile of TGc-AUC in lean subjects, was 30.7 mmol.1 h/L and corresponded with a fasting TGc of 1.8 mmol/L (eg, 1.6 mmol/L in plasma), calculated using univariate regression analysis. In conclusion, daylong triglyceridemia is similarly increased in diabetics and obese nondiabetics compared with lean subjects. Fasting TG and central obesity largely determine daylong triglyceridemia, independent of the presence of type 2 diabetes. Decreasing fasting plasma TG below 1.6 mmol/L could lead to a normalization of postprandial lipemia in obese subjects with and without diabetes. PMID- 12898472 TI - Comparison of the priming effects of pulsatile and continuous insulin delivery on insulin action in man. AB - Insulin is normally secreted in man in regular pulses every 5 to 15 minutes. Disordered pulsation has been demonstrated in several insulin-resistant states and it is unclear whether this represents a primary beta-cell defect contributing to impairment of peripheral insulin action or rather is a consequence of insulin resistance. Basal or near basal insulin administration by pulsatile infusion augments hypoglycemic effect and improves insulin-mediated glucose uptake compared with insulin by continuous infusion. To date no study has examined whether normal basal insulin pulsatility is required to preserve subsequent insulin sensitivity during hyperinsulinemia. We studied the effect of overnight pulsatile versus continuous basal insulin on a subsequent hyperinsulinemic euglycemic clamp. Nineteen normal volunteers (male:female ratio, 17:2; mean age +/- SEM, 26.1 +/- 2.3 years) were studied on 2 occasions each. Endogenous insulin secretion was inhibited by octreotide (0.43 microg kg(-1). h(-1)) and replaced overnight at 5.4 mU kg(-1). h(-1) either by continuous infusion or in 2-minute pulses every 13 minutes (n = 10) or every 7 minutes (n = 9). Glucagon was replaced at physiological concentration by continuous infusion (30 ng. kg(-1). h( 1)). Venous plasma glucose overnight was not significantly different between the pulsatile and continuous protocols. After discontinuing the overnight insulin infusion, insulin action was assessed during a hyperinsulinemic euglycemic clamp (1 mU kg(-1). h(-1)). Glucose infusion rates at steady-state during the hyperinsulinemic clamp were similar between continuous and both frequencies of pulsatile infusion (continuous 44.6 +/- 4.3 micromol. kg(-1). min(-1) v 13-minute pulsatile 41.7 +/- 5.9 micromol. kg(-1). min(-1), P =.27; continuous 34.6 +/- 2.5 micromol. kg(-1) min(-1) v 7-minute pulsatile 41.4 +/- 3.2 micromol. kg(-1). min( 1), P =.08). We conclude that overnight pulsatile compared with continuous insulin administration has no different effect on subsequent peripheral insulin mediated glucose uptake. A priming effect cannot therefore explain the previously demonstrated association between endogenous insulin pulse frequency and peripheral insulin action. PMID- 12898473 TI - Preclinical diabetic cardiomyopathy: relation of left ventricular diastolic dysfunction to cardiac autonomic neuropathy in men with uncomplicated well controlled type 2 diabetes. AB - Diabetic cardiomyopathy is an ill-defined entity. This study was designed to explore the possible association between left ventricular diastolic dysfunction (LVDD) and cardiac autonomic neuropathy (CAN) independently from metabolic control. Three groups of 10 age-matched men each with well-controlled type 2 diabetes were studied: (1) subjects with normal diastolic function, (2) subjects with LVDD characterized by impaired LV relaxation, and (3) subjects with a more severe form of LVDD characterized by a pseudonormalized pattern of LV filling. No subject had evidence of clinical diabetic complications, coronary artery disease (CAD), hypertension, congestive heart failure, or thyroid or overt renal disease, and all had a negative maximal exercise test. LVDD was evaluated by Doppler echocardiographic and CAN was evaluated using spectral analysis of heart rate variability (HRV; time and frequency domains) from 24-hour Holter recordings. Findings showed that the high frequency power (HF: 0.15 to 0.4 Hz) tends to decrease with worsening diastolic function; 5.0 +/- 0.2 ms(2) (mean +/- SE) in group 1, 4.2 +/- 0.3 ms(2) in group 2, and 3.9 +/- 0.4 ms(2) (P =.03) in group 3, respectively, whereas the low frequency power (LF: 0.04 to 0.15 Hz) was similar between groups. In the time domain, the mean squared differences of the successive RR intervals (rMSDD) also showed the same pattern, ie, 31.0 +/- 2.8 ms, 23.8 +/- 1.6 ms, and 21.5 +/- 2.9 ms in groups 1, 2, and 3, respectively (P =.03). The E/A ratio correlated significantly with indices of parasympathetic modulation (HF; r = 0.448, P =.013; rMSDD: r = 0.457, P =.011; pNN50: r = 0.425, P =.019). LVDD and CAN are associated in patients with otherwise uncomplicated well-controlled type 2 diabetes. The parameters defining these 2 abnormalities may serve to better define diabetic cardiomyopathy as a distinct entity and could eventually become useful prognostic indicators as it has been shown in nondiabetic populations. PMID- 12898474 TI - Acute effects of decreased glutamine supply on protein and amino acid metabolism in hepatic tissue: a study using isolated perfused rat liver. AB - Glutamine deficiency, a common finding in severe illness, has a negative influence on immune status, protein metabolism, and disease outcome. In several studies, a close relationship between glutamine, branched-chain amino acid (BCAA), and protein metabolism was demonstrated. The aim of the present study was to investigate the effect of glutamine deficiency on amino acid and protein metabolism in hepatic tissue using a model of isolated perfused rat liver (IPRL). Parameters of protein metabolism and amino acid metabolism were measured using both recirculation and single pass technique with L-[1-(14)C]leucine and [1 (14)C]ketoisocaproate (KIC) as a tracer. Glutamine concentration in perfusion solution was 0.5 mmol/L in control and 0 mmol/L in the glutamine-deficient group. The net release of glutamine (about 11 micromol/g/h) and higher net uptake of most of the amino acids was observed in the glutamine-deficient group. There was an insignificant effect of lack of glutamine on hepatic protein synthesis, proteolysis, and the release of urea. However, significantly lower release of proteins by the liver perfused with glutamine-deficient solution was observed. The lack of glutamine in perfusion solution caused a significant decrease in leucine oxidation (6.66 +/- 1.04 v 13.67 +/- 2.38, micromol/g dry liver/h, P <.05) and an increase in KIC oxidation (163.7 +/- 16.5 v 92.0 +/- 12.9 micromL/g dry liver/h, P <.05). We conclude that decreased delivery of glutamine to hepatic tissue activates glutamine synthesis, decreases resynthesis of essential BCAA from branched-chain keto acids (BCKA), increases catabolism of BCKA, and has an insignificant effect on protein turnover in hepatic tissue. PMID- 12898475 TI - Lower rate of tumor necrosis factor-alpha -863A allele and higher concentration of tumor necrosis factor-alpha receptor 2 in first-degree relatives of subjects with type 2 diabetes. AB - Allelic variants of the tumor necrosis factor-alpha (TNF-alpha) gene seem to contribute to insulin resistance increasing the transcription rate of TNF-alpha. The TNF-alpha -863A allele is associated with a lower expression of TNF-alpha gene and less secretion of the cytokine. To investigate whether an abnormal TNF alpha system regulation may contribute to early impairment of insulin action in first-degree relatives of patients with type 2 diabetes mellitus (DM), we studied the TNF-alpha -863C/A polymorphism and the soluble fraction of TNF-alpha receptor 2 (sTNFR2) concentration in these subjects in comparison to a control group. A total of 52% of subjects in the relatives' group showed an abnormal oral glucose tolerance (either as impaired glucose tolerance [IGT] or diabetes) and had more features of the insulin resistance syndrome, despite showing similar body composition as controls. The plasma concentration of the sTNFR2 was higher and insulin sensitivity (%S) was lower in the relatives' group than in the controls. Likewise, the TNF-alpha -863A allele was more commonly detected in the control group (10 of 41) than in the relative's group (2 of 36, P =.029). In a multivariate linear regression analysis, neither TNF-alpha -863A allele nor sTNFR2 independently determined %S. Only body mass index (BMI) and the presence of a positive family history of DM were independent determinants of insulin resistance. In summary, our study showed a lower rate of TNF-alpha -863A allele and higher concentrations of sTNFR2 in first-degree relatives of DM subjects. These findings could be included among the genetic, metabolic, and clinical heterogeneity that characterizes the pathophysiology of DM. The presence of abnormalities in the TNF-alpha pathway could predispose to the development of DM in subjects at risk for the disease. PMID- 12898476 TI - Dietary sodium restriction exacerbates age-related changes in rat adipose tissue and liver lipogenesis. AB - To investigate the effects of prolonged dietary sodium restriction on lipid metabolism, male rats weighing 35 to 40 g (just weaned) were fed either a low salt (LSD) or a normal salt diet (NSD) and used in metabolic experiments after 1, 2, or 3 months of diet consumption. After 2 and 3 months on the diet, LSD rats showed increased amounts of lipid in carcass and retroperitoneal tissue. In both LSD and NSD, extending the feeding period from 2 to 3 months resulted in a marked reduction in the in vivo rates of adipose tissue fatty acid synthesis that was accompanied by increases in liver lipogenesis and in the activity of adipose tissue lipoprotein lipase (LPL). However, these increases were more marked in LSD rats. Thus, in vivo rates of liver fatty synthesis and LPL activity in LSD rats, which were already higher (by about 35% and 20%, respectively) than in controls after 2 months, attained levels 50% higher than those in NSD animals after another month on the diet. Brown adipose tissue (BAT) thermogenic capacity, estimated after 2 and 3 months by the tissue temperature response to norepinephrine (NE) injection and by guanosine diphosphate (GDP) binding to BAT mitochondria, did not change in controls, but was significantly reduced in LSD rats. This raises the possibility that a decrease in overall energy expenditure, together with an LPL-induced increased uptake of preformed fatty acids from the circulation, may account for the excessive lipid accumulation in LSD rats. Taken together, the data indicate that prolonged dietary sodium restriction exacerbates normal, age-related changes in white and BAT metabolism. PMID- 12898477 TI - Rosiglitazone improves muscle insulin sensitivity, irrespective of increased triglyceride content, in ob/ob mice. AB - The present study was performed to examine the effects of rosiglitazone treatment on tissue-specific insulin sensitivity. Therefore, we used obese, insulin resistant ob/ob mice and measured the effects of rosiglitazone treatment on insulin sensitivity and simultaneously tissue-specific uptake of glucose and free fatty acids (FFA) under hyperinsulinemic euglycemic clamp conditions. Rosiglitazone treatment resulted in significantly higher body weight and decreased plasma levels of glucose, insulin, and triglyceride (TG). Glucose tolerance, as well as insulin sensitivity, was improved upon rosiglitazone treatment, as assessed by glucose tolerance and insulin sensitivity tests. Hyperinsulinemic euglycemic clamps showed increased glucose infusion rates with increased whole body insulin sensitivity. Rosiglitazone treatment resulted in increased glucose uptake by cardiac and skeletal muscle under hyperinsulinemic euglycemic clamp conditions, while no differences were observed in FA uptake. Measurement of TG content showed that rosiglitazone treatment resulted in decreased TG content of cardiac muscle, but increased TG content of skeletal muscle. We conclude that rosiglitazone treatment leads to strong improvement of insulin sensitivity, irrespective of increased muscle TG content, in ob/ob mice. PMID- 12898478 TI - Documentation of ED patient pain by nurses and physicians. AB - The purpose of this study was to evaluate ED documentation of patient pain in light of the Joint Commission of Accreditation of Healthcare Organization's emphasis on pain assessment and management. A prospectively designed pain management survey was offered to patients on ED discharge. Documentation of pain intensity by ED nurses and physicians was retrospectively reviewed. Of 302 patients surveyed, 261 (86%) complete charts were available for review. Initial pain assessments were noted on 94% of the charts, but a pain scale was used for only 23% of the patients. Documentation of pain subsequent to therapy was noted on 39% of the charts, but a pain scale was used only 19% of the time. Subsequent to therapy, nurses were 2.2 x more likely to document pain assessments than physicians (30% vs 16%, P <.001). Patients with severe pain on arrival (46% vs 31%, odds ratio [OR] = 1.9, P <.02), chest pain (72% vs 32%, OR = 5.4, P <.001), or those receiving powerful analgesics (62% vs 32%, 3.5, P <.001) were more likely to receive a documented subsequent pain assessment than other patients. Pain severity is not consistently documented in ED patients, especially after therapy has been provided. Patients with severe pain and those receiving powerful analgesics were more likely to have a pain assessment subsequent to ED therapy. PMID- 12898479 TI - Patterns of injury in hospitalized terrorist victims. AB - Acts of terror increase the demand for acute care. This article describes the pattern of injury of terror victims hospitalized at 9 acute-care hospitals in Israel during a 15-month period of terrorism. To characterize patients hospitalized as a result of terror injuries, we compared terror casualties with other injuries regarding severity, outcome, and service utilization. Using data from the National Trauma Registry, characteristics of casualties are portrayed. During the study period, 23,048 patients were recorded, 561 of them (2.4%) were injured through terrorist acts. Seventy percent were younger than 29 years. Seventy-five percent were males. Thirteen percent of terror victims compared with 3% with other traumatic injuries, arrived by helicopter. Injury mechanism consisted mainly of explosions (n = 269, 48%) and gunshot injuries (n = 266, 47%). One third of the population experienced severe trauma (Injury Severity Score > or = 16). One hundred-forty-two patients (26%) needed to be admitted to the intensive-care unit. Inpatient mortality was 6% (n = 35). Fifty-five percent of the injuries (n = 306) included open wounds and 31% (n = 172) involved internal injuries; 39% (n = 221) sustained fractures. Half of the patients had a procedure in the operating room (n = 298). Duration of hospitalization was longer than 2 weeks for nearly 20% of the population. Injuries from terrorist acts are severe and impose a burden on the healthcare system. Further studies of the special injury pattern associated with terror are necessary to enhance secondary management and tertiary prevention when occurring. PMID- 12898480 TI - Safety and delay time in prehospital thrombolysis of acute myocardial infarction in urban and rural areas in Sweden. AB - Sixteen hospitals in Sweden, including those in urban and more sparsely populated areas, and the associated ambulance organizations were enrolled in a prospective evaluation of the feasibility of treating patients with a ST-elevation infarction with a thrombolytic agent (reteplase) before hospital admission. A physician staffed the ambulances in 1% of cases, a nurse in 67%, and a staff nurse in 32% of cases. In all, 64 patients in urban areas and 90 patients in rural areas were included. The occurrence of complications before hospital admission was low and similar in the 2 groups. The median interval between the onset of symptoms and the start of thrombolysis was 1 hour 44 minutes in urban areas versus 2 hours 14 minutes in rural areas (P = 0.03). The median arrival time (interval between onset of symptoms and arrival of the ambulance) tended to be shorter in urban areas (1 hr 10 min vs 1 hr 33 min; not significant) and the median interval between the arrival of the ambulance and the start of thrombolysis was shorter in urban areas (27 min vs 36 min; P < 0.0001). When comparing urban areas with the least-populated rural areas, differences in various delay times became even more marked. Patients in urban areas had a higher ejection fraction and fewer symptoms of heart failure after 30 days and a lower 1-year mortality. PMID- 12898481 TI - Climatic and diurnal variation in suicide attempts in the ED. AB - The aim of this study was to determine whether there is a relationship between climatic factors and suicidal behavior. A total of 1,119 suicide attempts were collected from hospital records between 1996 and 2001. A clear seasonal variation was seen in suicide attempts in the 15-24, 25-34, and over 65 age groups in men and in the 15-24, 25-34, and 35-44 age groups in women with peaks in the spring and summer. Suicide attempts were more frequent between the hours of 6:00-9:00 pm in males and 3:00-6:00 pm in females. People attempting suicide who have depression, anxiety, or a psychotic disorder usually attempt suicide in the summer. Whereas the monthly averages of humidity, ambient temperature, duration and intensity of sunlight were positively correlated with the number of monthly suicide attempts, cloudiness and atmospheric pressure were negatively correlated. In conclusion, we must keep in mind that suicides and suicide attempts are not only the effect of climatic changes and that the most important component is the individual's ability to deal with conflicts. PMID- 12898482 TI - Variation in outcome in studies of out-of-hospital cardiac arrest: a review of studies conforming to the Utstein guidelines. AB - The objective of this study was to systematically review studies on out-of hospital cardiac arrest published according to the Utstein guidelines to describe the variability in factors of resuscitation and outcome. Articles that reported primary data on survival after out-of-hospital cardiac arrest in the Utstein style were included. Forty-seven articles were identified using Medline. Fourteen studies met our criteria for inclusion. The number of patients in whom resuscitation was attempted varied between 78 and 3,243. The proportion of bystander-witnessed cases varied between 38% and 89%; bystander CPR was performed in 21% to 56% of the cases. Patients with a bystander-witnessed cardiac arrest of cardiac etiology were discharged alive in 2% to 49% of the cases. Even when data are reported in a uniform way as suggested by the Utstein template, there is a tremendous variability in outcome. This did not appear to be entirely explained by variability in the traditional risk factors for a low chance of survival. One cannot exclude the possibility of other factors being of ultimate importance for the outcome. PMID- 12898483 TI - Impact of stress testing on 30-day cardiovascular outcomes for low-risk patients with chest pain admitted to floor telemetry beds. AB - The role of immediate stress testing in low-risk patients with a potential acute coronary syndrome has not been rigorously evaluated with respect to impact on 30 day cardiovascular events. We evaluated the impact of inpatient, outpatient, or no stress testing (ETT) on 30-day cardiovascular outcomes. We performed a prospective cohort study in which consecutive patients with chest pain were admitted to a non-intensive-care telemetry bed over 16 months. Patients were identified in the ED, followed daily through hospitalization, and contacted by telephone at 30 days. Patients were excluded if they were admitted to the coronary care unit, died during hospitalization, sustained an acute myocardial infarction (AMI), or received cardiac catheterization before ETT. Patients were stratified according to whether they received an ETT as an inpatient, outpatient, or no ETT. Main outcomes were 30-day cardiac death, AMI, percutaneous interventions (PCI), and coronary artery bypass graft surgery (CABG). Data are presented as percentages with 95% confidence intervals (CI) for main outcomes. A total of 832 patients were admitted 962 times. A total of 205 patients (21%) received an in-house ETT. Seventy-four patients (10%) without an inpatient ETT received an outpatient ETT. At baseline, the groups were similar with respect to likelihood of ischemia based on mean ACI-TIPI score and Goldman risk score. A total of 98% of patients had 30-day follow-up. The cardiovascular outcomes (with 95% confidence interval) for patients with inpatient ETT versus outpatient ETT versus no ETT were as follows: death, 0% (0-1.5%) vs 0% (0-4.1%) vs 1% (0.3 1.7%); AMI, 1% (0.1-2.4%) vs 1.4% (0.1-4.1%) vs 0.3% (0.1-0.7%); PCI, 0.5% (0.1 1.5%) vs 1.3% (0.1-4.1%) vs 0% (0-0.4%); and CABG, 0.5% (0.1-1.5%) vs 0% (0-4.1%) vs 0.2% (0.1-0.4%). There was no statistical difference in 30-day cardiovascular outcomes among patients who received inpatient, outpatient, or no ETT within 30 days. This suggests that patients with chest pain who are admitted to non intensive-care telemetry (or observation unit) beds might not need stress testing before hospital release. PMID- 12898484 TI - Patient compliance with medical advice given by telephone. AB - Data on compliance with medical advice given by telephone consultation services are currently lacking. The aim of this study was to assess patient compliance with medical advice given by a call center. A cross-sectional telephone survey was carried out on a random sample of 463 callers 72 hours after contacting the Grenoble Dial 15 center in France. Four hundred nine subjects (88.3%) participated in the study. Of these, 286 callers (69.9%) complied with the medical advice given. Compliance was 61.4% among patients who were advised to treat themselves, 83.9% among patients who were advised to consult a general practitioner during business hours, and 64.0% among patients who were advised to go to an accident and ED (P <.01). The survey pointed out adverse events resulting from the service. Assessing patient compliance can be an important source of information for improving aspects of patient management provided by telephone consultation services. PMID- 12898485 TI - Troponin T in elders with suspected acute coronary syndromes. AB - Troponin T (TnT) elevations (> or =0.1 ng/mL) indicate an increased risk of adverse outcomes in patients with acute coronary syndromes (ACS). There is little data on the prognostic significance of TnT in elders with ACS. We sought to define the ability of TnT to predict adverse outcomes in elders with suspected ACS compared with the younger cohort. This is a nested cohort study of patients aged <65 and > or =65 years with suspected ACS. Serial ECGs, CKMB and TnT assays were obtained at presentation and 4, 8, and 16 hours later. Adverse outcomes at hospital discharge included death, nonfatal myocardial infarction, coronary artery bypass grafting, and positive cardiac catheterization. A total of 695 patients were enrolled. A total of 301 (48%) were aged 65 or older. Although there was no difference in TnT sensitivity between the younger and older cohorts, there was a difference in specificity, 94% versus 83% (P <.01), respectively. In both cohorts, renal insufficiency was associated with a significantly lower TnT specificity. In both elders and younger patients with abnormal renal function, low TnT specificity warrants careful consideration of this marker as the sole criterion for aggressive medical management. PMID- 12898486 TI - Tetanus immunization shortage in the United States. AB - The purpose of this study is to determine the effect of the tetanus immunization shortage on EDs and the EPs understanding of the prioritization of persons needing tetanus immunization. A survey consisting of questions about knowledge of the tetanus shortage, prioritization of immunizations, incidence of tetanus infections, and understanding of CDC recommendations was mailed to a random sample 20% of the US ED medical directors. The results of the survey were input into the SPSS program (SPSS, version 10, Chicago, IL). The survey was returned by 618 of the 1,375 (44.9%) ED medical directors in the United States. Almost all (97.2%, 601 of 617) were notified about the tetanus shortage and 58.3% (360 of 617) reported a shortage. A total of 42.2% (199 of 472) gave tetanus toxoid (TT), instead of tetanus and diphtheria toxoids adult type (dT) when indicated. Only 11.6% of those surveyed (56 of 482) established a patient callback system. Routine vaccination was stopped in 37.5% of the reporting hospitals, most often for adults and children (57.5%, 69 of 120). Twelve hospitals (1.9%) reported they had an increase in tetanus. Although 87.5% of the respondents (539 of 616) stated they were familiar with the CDC's prioritization for tetanus immunization, only 1.8% (11 of 616) got the prioritization correct. Although EM directors uniformly know about and are experiencing the tetanus shortage, few correctly reported the tetanus immunization priority. Few EDs had a patient callback system. PMID- 12898487 TI - Early use of glycoprotein IIb/IIIa inhibitors in the ED treatment of non-ST segment elevation acute coronary syndromes: a local quality improvement initiative. AB - A prospective observational study was conducted in 2,007 patients experiencing chest pain to determine impact of local quality improvement (QI) measures on the use of glycoprotein (GP) IIb/IIIa inhibitors in the ED treatment of high-risk patients with non-ST-segment elevation acute coronary syndromes (ACS). Patients with injury on the initial ECG or new sustained injury on continuous ECG were excluded. QI interventions were as follows: control (0-4 mo): no interventions (standardized protocols and prewritten orders in place 4 months prior); phase I (5-8 mo): simple education/awareness program with posted drug information pamphlets and eligibility criteria; phase II (9-12 mo): mandated QI form with real-time feedback and focused one-on-one physician education championed by an ED physician QI advocate. A total of 179 (8.9%) of the study patients met predefined high-risk criteria. Of these, a total of 41 (23.0%) patients had GP IIb/IIIa inhibitor therapy initiated in the ED. Percent of high-risk patients receiving therapy increased from 6.0% during the control phase to 16.1% during phase I and 50.9% during phase II. After controlling for patient demographics, patients treated during phase I had a 2.8 times increased odds (95% confidence interval CI: 0.8-10.3; P =.11 [not significant]) of receiving GP IIb/IIIa inhibitor relative to the control phase, and patients treated during phase II had a 20.2 times increased odds (95% CI: 6.1-66.9; P <.0001) of treatment. In conclusion, local QI measures incorporating standardized protocols, preprinted orders, physician education, and interactive feedback championed by an ED QI physician advocate can increase early use of GP IIb/IIIa inhibitors in the ED treatment of high-risk patients presenting with chest pain. PMID- 12898488 TI - HIV postexposure prophylaxis practices by US ED practitioners. AB - To determine how often US ED practitioners have prescribed HIV post-exposure prophylaxis (HIV PEP) and to discern how willing they are to offer it to patients, the authors surveyed 600 ED practitioners attending a national conference. According to their self-report, 11% had taken HIV PEP themselves. Sixty-eight percent had prescribed HIV PEP at some time. Of these, 92% had treated needlestick-injured health care workers, 48% sexual assault survivors, and 49% nonhealth care needlestick-injured persons. ED practitioners were more willing to offer HIV PEP after exposures to HIV-infected or high-risk sources than unknown or low- risk sources, as well as after sexual assault than consensual sex. Female practitioners, those who had themselves taken HIV PEP, resident physicians, and ED practitioners with fewer than 6 years of clinical practice were generally more apt to offer HIV PEP. Educational campaigns appear to be necessary to help ED practitioners determine when HIV PEP is appropriate. PMID- 12898489 TI - The effect of ED prescription dispensing on patient compliance. AB - The objective of this study was to evaluate whether dispensing prescriptions in the ED affects patient compliance and return visits to the hospital. Seventy-four patients who were deemed suitable candidates for outpatient therapy with a macrolide antibiotic were identified and prospectively randomized to receive either an entire course of azithromycin from the ED or a prescription for azithromycin to be filled at a local pharmacy free of charge. Pharmacy records and telephone interview were used to measure compliance with patients. Significantly fewer patients filled their prescription in the pharmacy group (74.2%) compared with the ED group, in which all patients received their medication. However, there was no difference in the self-reported compliance of completing the entire course of antibiotics between patients in the ED group (94.3%) and in the pharmacy group (96.8%). There was no significant difference between groups in return ED visits or hospital admissions. We conclude that delivery of prescriptions in the ED significantly increases the likelihood that the patient will obtain the medication prescribed. Whether the patients actually take the medication as directed is unknown. Patient's self-report did not accurately reflect true compliance and more objective means for measuring compliance is warranted. PMID- 12898490 TI - Effect of physostigmine and gastric lavage in a Datura stramonium-induced anticholinergic poisoning epidemic. AB - This study examines the impact of the administration of physostigmine and of nasogastric evacuation of Jimsonweed seeds on intensive-care unit (ICU) use and the length of stay in the hospital after Jimsonweed poisoning. Clinical data for this retrospective study were gathered from records of consecutive patients treated for Jimsonweed poisoning from September to November 1997. Descriptive statistics, Fisher's exact test, and Student t-test were used to analyze important clinical and sociodemographic variables. There were 17 victims of the Jimsonweed ingestion epidemic, all of whom presented with an anticholinergic toxidrome 3 to 9 hours after ingestion. Reported quantities of seed ingestion ranged from a low of 7 seeds to as high as 200 seeds. Altered mentation, manifested by combative behavior, necessitated admission of 13 patients to the ICU. The administration of physostigmine did not reduce admissions to the ICU (P = 0.54) or reduce length of stay in the hospital (P = 0.45) compared with the use of benzodiazepines alone. Nasogastric lavage was performed in 14 (82%) and seeds were recovered in 8 (57%) of those lavaged. The successful removal of Jimsonweed seeds did not decrease ICU use rates (P = 0.68) or shorten length of stay in the hospital compared with not recovering seeds (P = 0.85). The use of physostigmine and the successful nasogastric lavage of Jimsonweed seeds did not result in decreased intensive-care use or shorter length of stay in the hospital for Jimsonweed-induced anticholinergic toxicity. PMID- 12898491 TI - Care of children who have had surgery for congenital heart disease. AB - Children who have had surgical correction for congenital heart disease can present to the ED with an acute illness that could be associated with their cardiac lesion. There is no data available to summarize complications that could be associated with surgically corrected congenital heart disease. This work was undertaken to describe the common procedures used, list known complications of these procedures, and review general management principles in caring for the acutely ill child who has had heart surgery. PMID- 12898492 TI - Scurvy: historical review and current diagnostic approach. AB - Scurvy, a deficiency of vitamin C, now most often occurs in disadvantaged groups seen frequently in EDs: alcoholics with poor nutrition, the isolated elderly, and the institutionalized. Its prominent clinical features are lethargy; purpuric lesions, especially affecting the legs; myalgia; and, in advancing disease, bleeding from the gums with little provocation. Common misdiagnoses are vasculitis, blood dyscrasias, and ulcerative gingivitis. Untreated, scurvy is inevitably fatal as a result of infection or sudden death. Fortunately, individuals with scurvy, even those with advanced disease, respond favorably to administration of vitamin C. PMID- 12898493 TI - A visual schematic for clarifying the temporal relationship between the anion and osmol gaps in toxic alcohol poisoning. AB - Delayed treatment of ethylene glycol or methanol poisoning can result in life threatening complications, but obtaining quantitative levels is delayed by several hours in most hospitals. Calculating the anion gap and the osmol gap are recommended to hasten identification and treatment in cases of suspected toxic alcohol poisoning. However, relying on the anion and osmol gap calculations without understanding the temporal relationship between these 2 gaps can lead to delayed identification and increased morbidity in cases of toxic alcohol poisoning. Our "Mountain" schematic illustrates the presence of an elevated osmol gap found early in toxic alcohol poisoning and the presence of an elevated anion gap found later in the course of poisoning. Using the "Mountain" diagram as a clarification of the temporal relationship between the anion and the osmol gap can improve the diagnostic use of these screening assays. PMID- 12898494 TI - Avascular necrosis of common bones seen in the ED. AB - Missed diagnosis of avascular necrosis (AVN) may result in substantial morbidity. Early diagnosis is crucial for appropriate intervention and, ultimately, improved outcome. Emerging physicians need to recognize AVN to avoid unsatisfactory clinical results for their patients. Avascular necrosis (AVN) of the bone can occur when the blood supply to the bone is disrupted and is usually found in areas with terminal circulation. Commonly involved bones include the femoral head, talus, and scaphoid. PMID- 12898495 TI - Diagnosing thyrotoxic periodic paralysis in the ED. AB - Thyrotoxic periodic paralysis (TPP) and sporadic periodic paralysis (SPP) are the most common causes of hypokalemic periodic paralysis (HPP) in EDs in Asia. Their neuromuscular presentations are almost indistinguishable. We conducted this study to identify clinical clues that can help EPs distinguish between TPP and SPP. Thirty-four patients presenting to the ED with HPP were enrolled during a 3-year period. They did not have known hyperthyroidism before the attack and no family history of paralysis. They all had low K(+) excretion rates. Vital signs and blood biochemistry, including acid-base and electrolytes, were measured. TPP was subsequently established by thyroid function tests. Twenty patients had TPP and 14 patients had SPP. There was no significant difference in age and sex distribution between them. Systolic (SBP) but not diastolic blood pressure (SBP 145 +/- 4 vs 128 +/- 4 mm Hg, P < 0.001) and heart rate (106 +/- 3 vs 73 +/- 3 beats/min, P < 0.001) were significantly higher in those experiencing TPP than SPP. Among the biochemical factors, only plasma phosphate concentration (2.2 +/- 0.2 vs 3.2 +/- 0.2 mg/dL, P < 0.001) was significantly lower in those experiencing TPP than SPP. Systolic hypertension, tachycardia, and hypophosphatemia are clinical clues favoring the diagnosis of TPP. PMID- 12898496 TI - Spontaneous hemopneumothorax: an overlooked life-threatening condition. AB - Spontaneous hemopneumothorax is a rare clinical entity. A large spontaneous hemopneumothorax is life-threatening, and mortality increases with delayed recognition and intervention. The initial chest radiography and the amount of blood drained from the inserted chest tube frequently underestimates the actual blood loss from the active bleeder around the ruptured apical bullae, leading to failed recognition of a potentially life-threatening condition until unexpected hemodynamic collapse develops. We report 2 cases of spontaneous hemopneumothorax to emphasize the importance of early recognition and prompt surgical intervention by video-assisted thoracoscopic surgery (VATS). PMID- 12898497 TI - Management of patients with unstable angina and non-ST-segment elevation myocardial infarction: update ACC/AHA guidelines. PMID- 12898500 TI - Evaluation of a mental health treatment court with assertive community treatment. AB - Without active engagement, many adults with serious mental illnesses remain untreated in the community and commit criminal offenses, resulting in their placement in the jails rather than mental health facilities. A mental health treatment court (MHTC) with an assertive community treatment (ACT) model of case management was developed through the cooperative efforts of the criminal justice and mental health systems. Participants were 235 adults with a serious mental illness who were booked into the county jail, and who volunteered for the study. An experimental design was used, with participants randomly assigned to MHTC or treatment as usual (TAU), consisting of adversarial criminal processing and less intensive mental health treatment. Results were reported for 6 and 12 month follow-up periods. Clients in both conditions improved in life satisfaction, distress, and independent living, while participants in the MHTC also showed reductions in substance abuse and new criminal activity. Outcomes are interpreted within the context of changes brought about in the community subsequent to implementation of the MHTC. PMID- 12898501 TI - Perspectives on probation and mandated mental health treatment in specialized and traditional probation departments. AB - Despite the prevalence of mentally ill probationers, and probation officers' (POs') central role in their supervision, this is the first reported study to investigate how POs implement mandates to participate in psychiatric treatment. Five focus groups were conducted in major cities with 32 POs and 20 probationers representing a mix of traditional and "specialty" probation agencies. Three key findings resulted. First, there were considerable differences between POs in specialty and traditional agencies in the nature, range, and timing of strategies applied to monitor and enforce treatment compliance. Second, the quality of PO probationer relationships colored POs' use of these strategies and was perceived as central to probationer outcomes. Relationships characterized by a respectful, personal, approach were perceived as more effective in achieving desired outcomes than those that were more authoritarian. Third, specialty agencies strongly emphasized offender rehabilitation whereas traditional agencies focused more exclusively on community safety. These agencies differed in how well probationers with mental illness "fit" their standard operating procedure. Implications for future research and directions for probation practice are discussed. PMID- 12898502 TI - Does fear of coercion keep people away from mental health treatment? Evidence from a survey of persons with schizophrenia and mental health professionals. AB - Mental health consumer advocates have long argued that involuntary treatment frightens persons with mental disorder and thus deters them from voluntarily seeking help. We surveyed 85 mental health professionals and 104 individuals with schizophrenia spectrum conditions to assess their experience with and perceptions of involuntary treatment and other treatment mandates. Of the clinicians, 78% reported that overall they thought legal pressures made their patients with schizophrenia more likely to stay in treatment. Regarding involuntary outpatient commitment, 81% of clinicians disagreed with the premise that mandated community treatment deters persons with schizophrenia from seeking voluntary treatment in the future. Of the consumer sample, 63% reported a lifetime history of involuntary hospitalization, while 36% reported fear of coerced treatment as a barrier to seeking help for a mental health problem-termed here "mandated treatment-related barriers to care." In bivariate analyses, reluctance to seek outpatient treatment associated with fear of coerced treatment (mandated treatment-related barriers to care) was significantly more likely in subjects with a lifetime history of involuntary hospitalization, criminal court mandates to seek treatment, and representative payeeship. However, experience with involuntary outpatient commitment was not associated with barriers to seeking treatment. Recent reminders or warnings about potential consequences of treatment nonadherence, recent hospitalization, and high levels of perceived coercion generally were also associated with mandated treatment-related barriers to care. In multivariable analyses, only involuntary hospitalization and recent warnings about treatment nonadherence were found to be significantly associated with these barriers. These results suggest that mandated treatment may serve as a barrier to treatment, but that ongoing informal pressures to adhere to treatment may also be important barriers to treatment. PMID- 12898503 TI - Effects of involuntary outpatient commitment on subjective quality of life in persons with severe mental illness. AB - Recent evidence suggests that involuntary outpatient commitment (OPC), when appropriately applied, can improve adherence with psychiatric treatment, decrease hospital recidivism and arrests, and lower the risk of violent behavior in persons with severe mental illness. Presumably these are benefits that improve quality of life (QOL); however, insofar as OPC involves legal coercion, the undesirable aspects of OPC could also exert a negative effect on quality of life, thus offsetting clinical benefits. Involuntarily hospitalized subjects, awaiting discharge under outpatient commitment, were randomly assigned to be released or continue under outpatient commitment in the community after hospital discharge, and were followed for one year. Quality of life was measured at baseline and 12 months follow-up. Treatment characteristics and clinical outcomes were also measured. Subjects who underwent longer periods of outpatient commitment had significantly greater quality of life as measured at the end of the 1 year study. Multivariable analysis showed that the effect of OPC on QOL was mediated by greater treatment adherence and lower symptom scores. However, perceived coercion moderated the effect of OPC on QOL. Involuntary outpatient commitment, when sustained over time, indirectly exerts a positive effect on subjective quality of life for persons with SMI, at least in part by improving treatment adherence and lowering symptomatology. PMID- 12898504 TI - The reported prevalence of mandated community treatment in two Florida samples. AB - Questions on multiple modes of mandated community treatment (e.g. outpatient commitment, advance directive, representative payee, and special housing) were included in two studies of persons with mental illness, using a mail survey of Medicaid enrollees receiving SSI and interviews conducted as part of an evaluation of a specialty mental health court. Results indicate that the majority of individuals reported no experience with any forms of mandated community treatment. However, respondents from the two samples who had been subject to community mandates reported comparatively similar experiences. Additional studies of the prevalence of mandated community treatment are necessary to garner expanded information regarding the use of such mandates. In addition, future studies must assure that the language employed to query individuals regarding such mandates is clearly understood by respondents, as certain terms (e.g. "outpatient commitment") may mean different things to different respondents. PMID- 12898505 TI - Waking Rip van Winkle: why developments in the last 20 years should teach the mental health system not to use housing as a tool of coercion. AB - Many housing programs for people with mental illnesses rely on models that require the person to adhere to treatment as a condition of continuing access to housing. These models that 'bundle' housing and treatment are relics of a past in which persons with mental illnesses were afforded little real choice in treatment, housing and other social supports. Conditioning access to housing in this manner is coercive and at odds with current thinking regarding treatment, as well as legal principles that shape the environment in which treatment is provided. This article summarizes the reasons why housing for people with mental illnesses should be provided free of the use of coercion. PMID- 12898506 TI - The antecedents of aggressive behavior among men with schizophrenia: a prospective investigation of patients in community treatment. AB - The association between psychotic symptoms and violence is unclear, due in part to methodological features of investigations that have examined this question, and in part to the fact that the association likely differs by disorder and treatment conditions. Using data from The Comparative Study of the Prevention of Crime and Violence by Mentally Ill Persons, we examined 128 men with schizophrenia or schizoaffective disorder discharged from general and forensic psychiatric hospitals in Canada, Finland, Germany, and Sweden. The association between symptoms and aggressive behavior was studied during two 6 month periods when the patients lived in the community. Severe positive and negative symptoms of psychosis, depression, and anxiety were measured at the beginning of each of the 6 month periods. In addition, at the beginning of the second 6 month period changes in symptoms in the previous period were indexed. Aggressive behavior was measured in each 6 month period by reports from patients and from collaterals. During the first 6 months post-discharge, after controlling for the presence of antisocial personality disorder or PCL score and past diagnoses of alcohol/drug abuse/dependence, the presence of a severe positive symptom significantly increased the risk of aggressive behavior. During the second 6 month period, after controlling for antisocial personality disorder or PCL score and self reported alcohol/drug use, the presence of a severe positive symptom, a TCO symptom, and an increase in TCO symptoms significantly increased the risk of aggressive behavior. Neither depot medications nor obligatory community treatment reduced the risk of aggressive behavior after controlling for the presence of a severe positive symptom and/or TCO symptoms. These findings suggest that, among men with schizophrenia being treated in the community, the presence of severe psychotic symptoms and the development of TCO symptoms are antecedents of aggressive behavior. PMID- 12898507 TI - TNF-alpha induction of lipolysis is mediated through activation of the extracellular signal related kinase pathway in 3T3-L1 adipocytes. AB - Tumor necrosis factor-alpha (TNF-alpha) increases adipocyte lipolysis after 6-12 h of incubation. TNF-alpha has been demonstrated to activate mitogen-activated protein (MAP) kinases including extracellular signal-related kinase (ERK) and N terminal-c-Jun-kinase (JNK) in different cell types. To determine if the MAP kinases have a role in TNF-alpha-induced lipolysis, 3T3-L1 adipocytes were treated with the cytokine (10 ng/ml), in the presence or absence of PD98059 or U0126 (100 micromoles), specific inhibitors of ERK activity. We demonstrated that U0126 or PD98059 blocked TNF-alpha-induced ERK activity and decreased TNF-alpha induced lipolysis by 65 or 76% respectively. The peroxisome-proliferator activated receptor gamma (PPARgamma) agonists, rosiglitazone (ros), and 15-deoxy Delta-(12,14)- prostaglandin J(2) (PGJ2) have been demonstrated to block TNF alpha-induced lipolysis. Pretreatment of adipocytes with these agents almost totally blocked TNF-alpha-induced ERK activation and reduced lipolysis by greater than 90%. TNF-alpha also stimulated JNK activity, which was not affected by PD98059 or PPARgamma agonist treatment. The expression of perilipin, previously proposed to contribute to the mechanism of lipolysis, is diminished in response to TNF-alpha treatment. Pretreatment of adipocytes with PD98059 or ros significantly blocked the TNF-alpha-induced reduction of perilipin A protein level as determined by Western analysis. These data suggest that activation of the ERK pathway is an early event in the mechanism of TNF-alpha-induced lipolysis. PMID- 12898508 TI - Jun N-terminal kinase pathway enhances signaling of monocytic differentiation of human leukemia cells induced by 1,25-dihydroxyvitamin D3. AB - Recent studies revealed that the MEK/ERK module of the mitogen-activated protein kinase (MAPK) signaling cascades is up-regulated in the early stages of 1alpha,25 dihydroxyvitamin D(3) (1,25D(3))-induced monocytic differentiation of human leukemia cells HL60. In the present study, we investigated whether another MAPK module, the JNK pathway, also participates in this form of differentiation. We found that the dependence on the concentration of the inducer, the vitamin hormone 1,25D(3), in two types of human leukemia cells, HL60 and U937, and the kinetics of monocytic differentiation in HL60 cells, parallel the degree of the activation of the JNK pathway. A blockade of JNK signaling by a stable expression of dominant negative (dn) JNK1 mutant in U937 cells resulted in reduced c-jun phosphorylation, and the differentiation of these cells was markedly decreased. Similarly, inhibition of JNK1 and JNK2 activities by the selective inhibitor SP600125 led to both dose-dependent reduction of c-jun and ATF-2 phosphorylation, and of the differentiation of HL60 cells. In addition, we found that JNK activity is essential for the AP-1 DNA binding induced by 1,25D(3) in HL60 and U937 cells. The results indicate that in cultured human leukemia cells, the JNK pathway participates in the induction of monocytic differentiation by 1,25D(3), probably by activating the AP-1 transcription factor. PMID- 12898509 TI - Pro-apoptotic activity of transiently expressed BCL-2 occurs independent of BAX and BAK. AB - BCL-2 suppresses apoptosis induced by a wide variety of stimuli in multiple cell types. Most of the in vitro studies that have examined the activity of BCL-2 have employed stable cell lines that ectopically express BCL-2. We have reported that BCL-2 is expressed at high levels in the absence of the 5'- and 3'-UTRs of the Bcl-2 gene and transient high level of expression results in potent cell death (Uhlmann et al., [1998]: JBC 278:17926-17932). Expression of BCL-2 under the transcriptional control of the cognate 5'- and 3'-UTRs express lower levels of BCL-2 and does not cause cell death. Our present results suggest that in contrast to BCL-2, transient expression of BCL-xL does not induce cell death and coexpression of BCL-xL with the pro-apoptotic BCL-2 does not suppress cell death. The pro-apoptotic activity of BCL-2 appears to involve activation of the cytochrome c/caspase 9/caspase 3 pathway. Elevated levels of BCL-2 expression results in N-terminal cleavage of BCL-2 at a novel site different from a previously identified caspase cleavage site at Asp 34 by a non-caspase protease. Transient expression of a BCL-2 mutant lacking aa 51-85 within the loop region induces efficient cell death and N-terminal cleavage of BCL-2 while a different deletion mutant lacking aa 30-91 induces reduced levels of cell death in the absence of BCL-2 cleavage suggesting that N-terminal processing of BCL-2 may be an amplification event in BCL-2-mediated cell death. Overexpression of BCL-2 in a Bax-null human colon cancer cell line (HCT116Bax-/-) induces efficient cell death. The pro-apoptotic activity of BCL-2 is also observed in a Bax-null cells in which BAK expression is inhibited by stable RNAi expression. Our results suggest that BCL-2 contains an intrinsic pro-apoptotic activity and can induce apoptosis independent of BAX and BAK under specific conditions. PMID- 12898510 TI - Merosin (laminin-2/4)-driven survival signaling: complex modulations of Bcl-2 homologs. AB - We have shown previously that the promotion of myofiber survival by the basement membrane component merosin (laminin-2 [alpha2beta1gamma1]/laminin-4 [alpha2beta2gamma1]) is dependent on the activity of the tyrosine kinase Fyn, whereas myofiber anoikis induced by merosin deficiency is dependent on the stress activated protein kinase p38alpha. To further understand such merosin-driven survival signaling, we analyzed the expression of five Bcl-2 homologs (Bcl-2, Bcl X(L), Bax, Bak, Bad) and one non-homologous associated molecule (Bag-1) in normal and merosin-deficient myotubes, with or without pharmacological inhibitors for Fyn and p38. Herein, we report that (1) merosin deficiency induces anoikis and causes decreased Bcl-2, Bcl-X(L), and Bag-1 levels, increased Bax and Bak levels, and decreased Bad phosphorylation; (2) Bcl-2, Bcl-X(L), Bag-1, and Bad phosphorylation are also decreased in anoikis-dying, Fyn-inhibited myotubes; (3) the inhibition of p38alpha in Fyn-inhibited and/or merosin-deficient myotubes protects against anoikis and increases Bcl-2 levels above normal, in addition to restoring Bad phosphorylation and Bag-1 levels to normal; (4) the overexpression of merosin in deficient myotubes also rescues from anoikis and increases Bcl-2 levels and Bad phosphorylation above normal, in addition to restoring Bcl-X(L), Bag-1, Bax, and Bak levels to normal; and (5) Bcl-2 overexpression is sufficient to rescue merosin-deficient myotubes from anoikis, even though the expression/phosphorylation levels of the other homologs analyzed are not restored to normal. These results indicate that merosin-driven myofiber survival signaling affects complex, differential modulations of individual Bcl-2 homologs. These further suggest that Bcl-2 can play a major role in suppressing myofiber anoikis. PMID- 12898511 TI - Epiregulin is more potent than EGF or TGFalpha in promoting in vitro wound closure due to enhanced ERK/MAPK activation. AB - Epiregulin (EPR) is a broad specificity EGF family member that activates ErbB1 and ErbB4 homodimers and all possible heterodimeric ErbB complexes. We have previously shown that topical EPR enhances the repair of murine excisional wounds. The purpose of this study was to determine whether EPR was more effective than EGF or TGFalpha in promoting in vitro wound closure and to compare the EPR induced signal transduction pathways with those activated by EGF and TGFalpha. Normal human epidermal keratinocytes or A431 cells were scratch wounded and treated for 24 h with varying doses of EPR, EGF or TGFalpha. Five-fold lower doses of EPR were significantly better than EGF or TGFalpha in stimulating in vitro wound closure. Mitomycin-c reduced EPR induced wound closure by 59%, versus a 9% and 25% decrease in EGF and TGFalpha induced closure. The ERK/MAPK inhibitor PD-98059 decreased EPR induced wound closure by 88%. By contrast, the PLC inhibitor U-73122, only reduced the EPR induced response by 21%. Immunoblot analysis revealed that 2 nM EPR stimulated a six-fold increase in p-ERK1/2, whereas 10 nM EGF or TGFalpha stimulated only a 3- and 2.5-fold increase in p ERK1/2. When compared with EGF or TGFalpha, EPR is a more potent and more effective inducer of in vitro wound closure due to its ability to promote significantly greater ERK/MAPK activation. PMID- 12898512 TI - Down-regulation of Flt-1 gene expression by the proteasome inhibitor MG262. AB - The mechanisms involved in the anti-angiogenic actions of the proteasome inhibitors are poorly understood. Here, we report that the gene expression of the VEGF receptor Flt-1 (vascular endothelial growth factor receptor 1) was down regulated by the reversible proteasome inhibitor MG262 in explant cultures of the developing chicken pecten oculi, a vascular organ consisting of endothelial cells, pericytes, and macrophages. In addition, the inhibitor prevented the induction of Flt-1 by lipopolysaccharide (LPS) in macrophages and down-regulated the expression of Flt-1 after LPS induction. Flt-1 gene expression was also down regulated by MG262 in cultures of human microvascular endothelial cells. Interestingly, a transcript of Flt-1, coding for a soluble form of the receptor (sFlt-1) with anti-angiogenic properties, was not down-regulated in the same extent. Only a small decrease in the expression of VEGF and Ang-2 was detected in the pecten oculi upon inhibition of the proteasome, while no major changes were observed in the expression of other angiogenic molecules, such as KDR or Ang-1. Since recent experiments have demonstrated the importance of anti-Flt-1 therapy in the inhibition of tumor angiogenesis, retinal angiogenesis, arthritis, and atherosclerosis (Luttun et al. [2002]: Nat Med 8:831-840), our observation on down-regulation of Flt-1 in microvascular endothelial cells and macrophages by MG262 supports the postulated role of the proteasome inhibitors as potential candidates for therapeutic modulation of angiogenesis and inflammation. PMID- 12898513 TI - TAK-778 enhances osteoblast differentiation of human bone marrow cells. AB - TAK-778 has been shown to induce bone growth in in vitro and in vivo models. However, there are no studies evaluating the effect of TAK-778 on human cells. Thus, the aim of this study was to investigate osteogenesis induced by TAK-778 on human bone marrow cells. Cells were cultured in 24-well culture plates at a cell density of 2 x 10(4) cells/well in culture medium containing TAK-778 (10(-7), 10( 6), and 10(-5) M, each) or vehicle. During the culture period, cells were incubated at 37 degrees C in a humidified atmosphere of 5% CO(2) and 95% air. For attachment evaluation, cells were cultured for 4 and 24 h. After 7, 14, and 21 days, cell proliferation, cell viability, total protein content, alkaline phosphatase (ALP) activity, and bone-like formation were evaluated. Data were compared by ANOVA and Duncan's multiple range test. TAK-778 did not affect cell attachment and viability. Cell number was reduced by TAK-778 in all time period evaluated in a dose-dependent way. The effect of TAK-778 on total protein content, ALP activity and bone-like formation was a dose-dependent increase. The present results suggest that initial cell events such as cell attachment are not affected by TAK-778 while events that indicate osteoblast differentiation including reduced cell proliferation, and increased both ALP activity and bone like formation are enhanced by TAK-778 in a time and dose-dependent way. It means that TAK-778 could be a useful drug to enhance new bone formation in clinical situations that require rapid restoration of physiologic function, such as orthopedic and maxillofacial surgery. PMID- 12898514 TI - Expression and localization of extracellular matrix metalloproteinase inducer in giant cell tumor of bone. AB - Matrix metalloproteinases (MMPs) are regarded as a significant regulator in tumor invasion and metastasis. Previous studies have shown that extracellular matrix metalloproteinase inducer (EMMPRIN) in tumor cells induces the synthesis of MMPs. EMMPRIN is abundantly present on the surface of tumor cells and stimulate adjacent stromal cells to synthesize MMPs to induce tumor progression. Giant cell tumor (GCT) of bone is a benign but locally aggressive primary neoplasm of bone. The spindle-shaped mononuclear stromal cells are considered to be the tumor components of GCT, which are capable of inducing osteoclast formation by recruiting the circulating monocyte and macrophage. In this study, we proposed that EMMPRIN is associated with the biological progression and aggressiveness of GCT. We have conducted semi-quantitative RT-PCR to determine the correlation of EMMPRIN expression with the clinical stage of GCT. We have also examined the cellular localization of EMMPRIN in GCT using in-situ hybridization (ISH) and Immunohistochemistry (IH). The results showed that EMMPRIN was present in GCT and its mRNA levels were associated with the clinical stage of GCT. Higher expression level of EMMPRIN was observed in GCT with advanced stage (stage III). There was a great significance (P < 0.05) of EMMPRIN expression between stage I & II and stage III GCTs. Both ISH and IH demonstrated that EMMPRIN is present at the multinuclear osteoclast-like giant cells of GCT, with strong immunostaining on the cell membrane. The stromal-like tumor cells were also positively stained but the intensity was weaker. Interestingly, the production of EMMPRIN in osteoclast like cells of GCT seems to be regulated by stromal-like tumor cells. Receptor activator of NF-kappaB ligand (RANKL), which has been previously shown to be produced by the stromal-like tumor cells for the recruitment of osteoclast-like giant cells in GCT, enhanced the expression of EMMPRIN mRNA during the differentiation of macrophage-like RAW(264.7) cells into osteoclasts. In short, our studies suggest that EMMPRIN may be an important regulatory factor involved in the biological behaviors of GCT. PMID- 12898515 TI - 9-cis retinoic acid accelerates calcitriol-induced osteocalcin production and promotes degradation of both vitamin D receptor and retinoid X receptor in human osteoblastic cells. AB - Abstract vitamin D receptor (VDR) and retinoid X receptor (RXR) heterodimerize to mediate the genomic actions of 1alpha,25-dihydroxyvitamin D(3) (1alpha,25(OH)(2)D(3), calcitriol), the biologically active form of vitamin D(3). In this study, we show that 9-cis retinoic acid (9-cisRA), the ligand for RXR, accelerates calcitriol-induced expression of osteocalcin gene, the marker for mature osteoblasts. Calcitriol and its synthetic analog KH1060 (1 nM) induced osteocalcin secretion after a 96-h incubation period as detected by radioimmunoassay. When these compounds were used together with 9-cisRA, osteocalcin protein secretion was, however, detected already after 72 and 48 h, respectively. Detection of osteocalcin mRNA with quantitative PCR revealed elevated mRNA levels already after a 4-h treatment of the cells with calcitriol, KH1060, or 9-cisRA compared with untreated cells. In combination treatments, 9 cisRA rapidly stimulated osteocalcin mRNA synthesis induced by the different vitamin D(3) compounds. In MG-63 cells treated with calcitriol or KH1060, the stimulation was maximal after the first 4 h and diminished thereafter. In fact, after the 48-h incubation 9-cisRA reduced osteocalcin mRNA levels in KH1060 treated cells, the amount of mRNA being only 44% of the levels obtained with KH1060 alone. The reduction was accompanied by an increased degradation rate of both VDR and RXRbeta in the presence of 9-cisRA. Furthermore, 9-cisRA increased the formation of RXRbeta-VDR-VDRE complex on the osteocalcin gene VDRE. These results suggest that 9-cisRA accelerates calcitriol-induced osteocalcin production in human osteoblastic cells through increased formation of transcriptionally active chromatin complexes and, subsequently, promotes degradation of the heterodimeric complex of VDR and RXR. PMID- 12898516 TI - Increased expression of Mcl-1 is responsible for the blockage of TRAIL-induced apoptosis mediated by EGF/ErbB1 signaling pathway. AB - Epidermal growth factor (EGF) protects against death receptor induced apoptosis in epithelial cells. Herein, we demonstrate that EGF protection against tumor necrosis factor related apoptosis-inducing ligand (TRAIL) induced apoptosis is mediated by increased expression of the Bcl-2 family member myeloid cell leukemia 1 (Mcl-1). EGF increased the mRNA and protein levels of Mcl-1. Furthermore, expression of ErbB1 alone or in combination with ErbB2 in NIH3T3 cells up regulates Mcl-1 following EGF treatment. In addition, up-regulation of Mcl-1 by EGF is mediated through AKT and NFkappaB activation since kinase inactive AKT and DeltaIkappaB effectively blocks this up-regulation. NFkappaB was also critical for the ability of EGF to prevent TRAIL induced apoptosis as a dominant negative IkappaB (DeltaIkappaB) blocked NFkappaB activation, and relieved EGF protection against TRAIL mediated mitochondrial cytochrome-c release and apoptosis. Finally, anti-sense oligonucleotides directed against Mcl-1 effectively reduced the protein levels of Mcl-1 and blocked EGF protection against TRAIL induced mitochondrial cytochrome-c release and apoptosis. Taken together, EGF signaling leads to increased Mcl-1 expression that is required for blockage of TRAIL induced apoptosis. PMID- 12898517 TI - Non-random features of loop-size chromatin fragmentation. AB - Upon isolation of DNA from normal eukaryotic cells by standard methods involving extensive proteolytic treatment, a rather homogeneous population of loop-size, double-stranded DNA fragments is regularly obtained. These DNA molecules can be efficiently end-labeled by the DNA polymerase I Klenow fragment, as well as by a 3'- to -5'-exonuclease-free Klenow enzyme, but not by terminal transferase (TdT) unless the ends have been filled up by Klenow, suggesting that dominantly 5' protruding termini are generated upon fragmentation. The filled-up termini were used for cloning the distal parts of the approximately 50 kb fragments. BLAST analysis of the sequence of several clones allowed us to determine the sequence of the non-cloned side of the breakpoints. Comparison of 25, 600 bp-long breakpoint sequences demonstrated prevalence of repetitive elements. Consensus motives characteristic of the breakpoint sequences have been identified. Several sequences exhibit peculiar computed conformational characteristics, with sharp transition or center of symmetry located exactly at the breakpoint. Our data collectively suggest that chromatin fragmentation involves nucleolytic cleavages at fragile/hypersensitive sites delimiting loop-size fragments in a non-random manner. Interestingly, the sequence characteristics of the breakpoints are reminiscent of certain breakpoint cluster regions frequently subject to gene rearrangements. PMID- 12898518 TI - LPS induces pulmonary intravascular macrophages producing inflammatory mediators via activating NF-kappaB. AB - Pulmonary intravascular macrophages (PIMs) are often responsible for the clearance of blood-borne pathogens, including endotoxin, lipopolysaccharide of Gram-negative bacteria. It is well accepted that PIMs play a pivotal role in the pathogenesis of endotoxin-induced acute lung injury. However, the mechanisms by which PIMs are involved in the lipopolysaccharide-induced inflammatory responses remain unclear. Through the present study the following results were found: (1) When challenged with lipopolysaccharide (10 micrograms/ml), PIMs underwent marked cellular enlargement, intercellular adhesion plaques became longer, and some particulates were enwrapped in the pseudopods. (2) Lipopolysaccharide could up regulate the expression of some inflammatory mediators in PIMs, including TNF alpha, IL-1beta, IL-6, IL-8, and COX-2, and these up-regulated expression of inflammatory mediators correlated with NF-kappaB activation. (3) Dexamethasone as well as acetylsalicylic acid reduced the expression of TNF-alpha in lipopolysaccharide-challenged PIMs, and the decreased expression of TNF-alpha was also consistent with decreased NF-kappaB activation. Our results suggest that NF kappaB activation in PIMs followed by phagocytizing lipopolysaccharide resulted in the up-regulation of TNF-alpha, IL-1beta, IL-6, IL-8, and COX-2, which could be alleviated by dexamethasone. PMID- 12898519 TI - alpha-cardiac actin (ACTC) binds to the band 3 (AE1) cardiac isoform. AB - The band 3 protein is the major integral protein present in the erythrocyte membrane. Two tissue-specific isoforms are also expressed in kidney alpha intercalated cells and in cardiomyocytes. It has been suggested that the cardiac isoform predominantly mediates the anion exchange in cardiomyocytes, but the role of the cytoplasmic domain of the band 3 (CDB3) protein in the cardiac tissue is unknown. In order to characterize novel associations of the CDB3 in the cardiac tissue, we performed the two-hybrid assay, using a bait comprising the region from leu 258 to leu 311 of the erythrocyte band 3, which must also be present in the cardiac isoform. The assay revealed two clones containing the C-terminal region of the alpha-cardiac actin. Immunoprecipitation of whole rat heart using an anti-actin antibody, immunoblotted with anti-human band 3, showed that actin binds to band 3 which was confirmed in the reverse assay. The confocal microscopy showed band 3 in the intercalated discs. Thus, besides the in vivo physical interaction in the Saccharomyces cerevisiae cell, we demonstrated using immunopreciptation that there is a physical association of band 3 with alpha cardiac actin in cardiomyocyte, and we suggest that the binding occur "in situ," in the intercalated disc, a site of cell-cell contact and attachment of the sarcomere to the plasma membrane. PMID- 12898520 TI - Non-apoptogenic killing of hela cervical carcinoma cells after short exposure to the alkylating agent N-methyl-N'-nitro-N-nitrosoguanidine (MNNG). AB - We examined the action of N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) on HeLa cells and compared it with that of cisplatin (CP). MNNG directly killed a substantial number of cells within 1 hour and resulted in strong DNA-damage as evidenced by Comet measurements. Despite appearance of DNA lesions, p53 protein was not activated. Analysis of HeLa cells treated with MNNG for 1h, 3h and 6h by flow cytometry and by Hoechst staining did not reveal any sub-G(1) cell population and chromatin condensation/fragmentation characteristic for apoptosis, respectively. Also, no biochemical changes typical for apoptosis such as activation of caspase-3 or release of cytochrome C from mitochondria were detected. Inactivation of PARP-1 reduced the direct cytotoxicity exerted by MNNG. Our results showing that despite appearance of severe DNA lesions after short exposure of HeLa cells to MNNG neither activation of p53 response nor induction of apoptosis occurred implicate that generation of strong DNA damage is not sufficient to stabilize p53 protein in HeLa cells. Our data unequivocally show that the conscientious determination of the type of cell death induced by genotoxic agents is necessary. The assessment of the changes based on at least a few independent criteria is required to discriminate between apoptosis and necrosis. Since the alkylating agents generate DNA strand breaks, the recruitment of methods based on determination of DNA cleavage such as DNA ladder or TUNEL assay for evaluation of apoptosis is not adequate. PMID- 12898521 TI - Characterization of mesenchymal stem cells isolated from murine bone marrow by negative selection. AB - Mesenchymal stem cells (MSCs) are typically enriched from bone marrow via isolation of the plastic adherent, fibroblastoid cell fraction. However, plastic adherent cultures elaborated from murine bone marrow are an admixture of fibroblastoid and hematopoietic cell types. Here we report a reliable method based on immunodepletion to fractionate fibroblastoid cells from hematopoietic cells within plastic adherent murine marrow cultures. The immunodepleted cells expressed the antigens Sca-1, CD29, CD44, CD81, CD106, and the stem cell marker nucleostemin (NST) but not CD11b, CD31, CD34, CD45, CD48, CD90, CD117, CD135, or the transcription factor Oct-4. They were also capable of differentiating into adipocytes, chondrocytes, and osteoblasts in vitro as well as osteoblasts/osteocytes in vivo. Therefore, immunodepletion yields a cell population devoid of hematopoietic and endothelial cells that is phenotypically and functionally equivalent to MSCs. The immunodepleted cells exhibited a population doubling time of approximately 5-7 days in culture. Poor growth was due to the dramatic down regulation of many genes involved in cell proliferation and cell cycle progression as a result of immunodepletion. Exposure of immunodepleted cells to fibroblast growth factor 2 (FGF2) but not insulin-like growth factor (IGF), murine stem cell factor, or leukemia inhibitory factor (LIF) significantly increased their growth rate. Moreover, 82% of the transcripts down regulated by immunodepletion remain unaltered in the presence of FGF2. Exposure to the later also reversibly inhibited the ability of the immunodepleted cells to differentiate into adipocytes, chondrocytes, and osteoblasts in vitro. Therefore, FGF2 appears to function as a mitogen and self-maintenance factor for murine MSCs enriched from bone marrow by negative selection. PMID- 12898522 TI - Co-culture of endothelial cells and smooth muscle cells affects gene expression of angiogenic factors. AB - Endothelial cells (EC) are in contact with the underlying smooth muscle cells (SMC). The interactions between EC and SMC in the vessel wall are considered to be involved in the control of growth and function of blood vessels. A co-culture system of EC and SMC and a method for separation of these cells was developed in order to investigate whether the presence of physical contact between EC and SMC affected the gene expression of angiogenic factors. Human EC and SMC were prepared from the great saphenous veins. Autologous EC were added on top of the confluent layer of SMC. After 72 h in co-culture, the EC were magnetically separated from SMC with the use of superparamagnetic beads. RT-PCR products for bFGF, bFGFR, VEGF, PDGF-AA, PDGF-BB, TGF-beta, and beta-actin were analyzed to study the mRNA expressions. The protein level of selected factors was studied by ELISA technique. In co-cultured SMC there was a statistically significant higher gene expression of VEGF, PDGF-AA, PDGF-BB, and TGF-beta and significant lower gene expression of bFGF and its receptor than in single cultured SMC. The protein level of PDGF-BB and TGF-beta was also significantly higher in co-cultured SMC. In co-cultured EC there were no significant differences in gene expression of PDGF-AA, PDGF-BB, and TGF-beta compared with single cultured EC. The gene expression and protein synthesis of VEGF was significantly higher in co-cultured EC. The findings from the present study suggest that cell-cell interactions of EC and SMC affect the gene and protein expression of angiogenic factors. PMID- 12898523 TI - Phosphorylation regulates the interaction and complex formation between wt p53 protein and PARP-1. AB - We recently characterized the interaction between poly(ADP-ribose) polymerase-1 (PARP-1) and the product of the tumor suppressor gene p53. We investigated which domains of human PARP-1 and of human wild-type (wt) p53 were involved in this protein-protein interaction. We generated baculoviral constructs encoding full length or distinct functional domains of both proteins. Full length PARP-1 was simultaneously coexpressed in insect cells with full length wt p53 protein or its distinct truncated fragments and vice versa. Reciprocal immunoprecipitation of Sf9 cell lysates revealed that the central and carboxy-terminal fragments of p53 were sufficient to confer binding to PARP-1, whereas the amino-terminal part harboring the transactivation functional domain was dispensable. On the other hand, the amino-terminal and central fragments of PARP-1 were necessary for complex formation with p53 protein. As the most important features of p53 protein are regulated by phosphorylation, we addressed the question of whether its phosphorylation is essential for binding between the two proteins. Baculovirally expressed wt p53 was post-translationally modified. At least six distinct p53 isomeres were resolved by immunoblotting following two-dimensional separation of baculovirally expressed wt p53 protein. Using specific phospho-serine antibodies, we identified phosphorylation of baculovirally expressed p53 protein at five distinct sites. To define the role of p53 phosphorylation, pull-down assays using untreated and dephosphorylated p53 protein were performed. Dephosphorylated p53 failed to bind PARP-1 indicating that complex formation between both proteins is regulated by phosphorylation of p53. The marked phosphorylation of p53 at Ser392 observed in unstressed cells suggests that the phosphorylated carboxy-terminal part of p53 undergoes complex formation with PARP-1 resulting in masking of the NES and thereby preventing its export. The functional significance of the interaction between both proteins was investigated at two different conditions: inactivation of PARP-1 and overexpression of PARP-1. Our results unequivocally show that the presence of PARP-1 regulates the basal expression of wt p53 in unstressed cells. PMID- 12898524 TI - Establishment of a T-Ag and NF-kappaB binding elements gene co-transfected stable HUVECs cell line. AB - To set up a T-Ag gene transfected stable human umbilical venous endothelial cells (HUVECs) cell line and a T-Ag, nuclear transcriptional factor kappa B (NF-kappaB) binding elements linked with luciferase reporter gene co-transfected stable HUVECs cell line. Cultured HUVECs were transfected with pCI-neo-T-Ag and pRSV-luc 3XkappaB by lipofectin. The G418 selected monoclones were subcultured. The expression of marker protein, vWF and the characteristic of uptake of lipids were compared by Western blotting and immunocytochemistry in non-transfected and transfected HUVECs. The reporter gene assay was done in the presence of TNF alpha. A T-Ag gene transfected stable HUVECs cell line and a T-Ag and NF-kappaB binding elements linked with luciferase reporter gene co-transfected stable HUVECs cell lines were set up. The expression of vWF of these cell lines was similar with those in non-transformed HUVECs. The function of uptaking of lipids was preserved as well in transfected cell lines. Furthermore, TNF-alpha, a typical cytokine increasing the activity of NF-kappaB was used to treat the transfected cells O/N. The higher luciferase reporter gene activity was seen. A pCI-neo-T-Ag and pRSV-luc-3X kappaB co-transfected stable HUVECs cell line might be used to check reporter gene activity directly. It might be a useful tool to screen drugs acting on transcription level. PMID- 12898525 TI - Effect of overexpressing fibroblast growth factor 2 protein isoforms in osteoblastic ROS 17/2.8 cells. AB - Fibroblast growth factor-2 (FGF-2) is made by osteoblasts and modulates their function. There are high molecular weight (HMW) protein isoforms of FGF-2 that have nuclear localization sequences and a low molecular weight (LMW) 18 kDa FGF-2 protein that is exported from cells. Since FGF-2 is a trophic factor and potent mitogen for osteoblasts, the goal of this study was to utilize targeted overexpression of FGF-2 as a novel means of assessing different FGF-2 isoforms on osteoblastic cell viability and proliferation. Either LMW or HMW human Fgf2 cDNAs were cloned downstream of 3.6 kb alpha1(I)-collagen 5' regulatory elements (Col 3.6). A set of expression vectors, called Col3.6-Fgf2 isoforms-IRES-GFPsaph, capable of concurrently overexpressing either LMW or HMW FGF-2 isoforms concomitant with GFPsaph from a single bicistronic mRNA were built. Viable cell number in ROS 17/2.8 cells stably transfected with Vector (Col3.6-IRES-GFPsaph) versus each of the Col3.6-Fgf2-IRES-GFPsaph constructs were compared. In the presence of 1 or 10% serum, DNA synthesis was increased in cells expressing any isoform of FGF-2 compared with vector. However, cells transfected with HMW isoform had augmented DNA synthesis in 1 or 10% serum compared with cells expressing either ALL or LMW FGF-2 isoforms. A neutralizing FGF-2 antibody significantly reduced the mitogenic response in cells harboring ALL or the LMW FGF-2 isoforms but did not block the mitogenic effect of cells harboring the HMW isoforms. In summary, overexpression of any isoform of FGF-2 protein increased viable cell number and OB proliferation in the presence of low or high concentrations of serum. However, the HMW/nuclear isoforms preferentially mediate augmented OB proliferation. We conclude that differential expression of FGF-2 proteins isoforms is important in modulating OB function. PMID- 12898527 TI - Expression and subcellular localization of Ror tyrosine kinase receptors are developmentally regulated in cultured hippocampal neurons. AB - Ror1 and Ror2 are two novel receptor tyrosine kinases that have been implicated in neuronal differentiation in Caenorhabditis elegans. As a first step toward elucidating their role in the mammalian brain, we analyzed their expression and localization patterns in hippocampal neurons. Our results showed that both receptors are expressed from early stages of development and that their protein levels peak during periods of active synapse formation. Immunocytochemical analysis indicated that Ror1 and Ror2 are highly concentrated in the growth cones of immature neurons and are present throughout the somatodendritic compartment of mature hippocampal cells. Further analysis indicated that they are present not only in the cell membrane but also in Triton- and saponin-insoluble fractions, suggesting that they may be associated with both the cytoskeleton and membrane bound organelles. Taken collectively, our results suggest that Ror1 and Ror2 might play a role during early stages of development in mammalian central neurons. PMID- 12898528 TI - Increased neurogenesis after experimental Streptococcus pneumoniae meningitis. AB - Neuronal damage in the hippocampal formation is a common feature in animal models of bacterial meningitis and human disease. In mouse and rabbit models of Streptococcus pneumoniae meningitis, proliferation of neural progenitor cells quantified by bromodeoxyuridine (BrdU) incorporation was enhanced in the subgranular layer of the dentate gyrus. In mice, the density of BrdU-labeled cells was maximal on Day 2 after infection. Approximately 60% of the cells labeled by BrdU between Days 7 and 10 after infection that remained present 28 days later had migrated into deeper layers of the dentate gyrus and differentiated into neurons, as evidenced by immunohistochemical staining for TUC 4, MAP-2 and beta-tubulin. This suggests that endogenous repair mechanisms may limit consequences of neuronal destruction after meningitis. PMID- 12898530 TI - Microarray analysis of gene expression in proliferating Schwann cells: synergistic response of a specific subset of genes to the mitogenic action of heregulin plus forskolin. AB - Cultured Schwann cells treated with heregulin growth factor require costimulation with a cyclic adenosine monophosphate-elevating agent to produce maximal cell proliferation. Gene chip expression analysis was used to identify genes that are induced or repressed when Schwann cells are treated with heregulin and/or forskolin. By utilizing arrays that contained 8799 probes, the expression of over 1000 genes was found to be significantly changed after 30 hr of treatment with heregulin, forskolin, or heregulin plus forskolin. Hierarchical clustering revealed groups of genes with distinct expression patterns. Of particular interest was a cluster of 140 genes that were up-regulated by heregulin plus forskolin but not by heregulin or forskolin alone. Many of the genes in this group have roles in cell division, such as cyclin B, cyclin D3, E2F-5, cdc 25B, polo-like kinase, and protein kinase C type III. These findings identify a profile of gene expression for Schwann cell proliferation. PMID- 12898529 TI - Alpha-tocopherol, an exogenous factor of adult hippocampal neurogenesis regulation. AB - In previous work, we found that adult hippocampal neurogenesis in rat is affected by vitamin E deficiency. Because vitamin E deficiency is a complex condition involving numerous biological systems, it is possible that its effect on postnatal new neuron production could be mediated by unknown changes in different factors that in turn play a role in this process. To clarify if vitamin E plays a direct role in regulating hippocampal neurogenesis, we studied the neurogenesis in adult control rats and in adult rats under supplementation with alpha tocopherol, the most important compound of vitamin E. The alpha-tocopherol level in control and supplemented rats was monitored. Qualitative and quantitative analysis of cell proliferation and death was carried out and expression of immature neuron markers PSA-NCAM, TUC 4, and DCX was investigated in hippocampus dentate gyrus. alpha-Tocopherol levels increased significantly in both plasma and brain after supplementation. Cell proliferation was inhibited in alpha-tocopherol supplemented rats, the number of dying cells was reduced, and the number of cells expressing the immature neuron markers was increased. The results obtained confirm and extend the idea that vitamin E is an exogenous factor playing a direct role in regulation of different steps of adult hippocampal neurogenesis. Some hypotheses about the possible mechanisms underlying the complex action of alpha-tocopherol, related to its antioxidant and molecule-specific non antioxidant properties, are proposed and discussed. PMID- 12898531 TI - Dysregulation of axonal sodium channel isoforms after adult-onset chronic demyelination. AB - Demyelination results in conduction block through changes in passive cable properties of an axon and in the expression and localization of axonal ion channels. We show here that adult-onset chronic demyelination, such as occurs in demyelinating disorders and after nerve injury, alters the complement of axonal voltage-dependent Na+ (Nav) channel isoforms and their localization. As a model, we used heterozygous transgenic mice with two extra copies of the proteolipid protein gene (Plp/-). Retinal ganglion cell axons in these mice myelinate normally, with young Plp/- and wild-type mice expressing Nav1.2 at low levels, whereas Nav1.6 is clustered in high densities at nodes of Ranvier. At 7 months of age, however, Plp/- mice exhibit severe demyelination and oligodendrocyte cell death, leading to a profound reduction in Nav1.6 clusters, loss of the paranodal axoglial apparatus, and a marked increase in Nav1.2. We conclude that myelin is crucial not only for node of Ranvier formation, but also to actively maintain the proper localization and complement of distinct axonal Nav channel isoforms throughout life. The altered Nav channel isoform localization and complement induced by demyelination may contribute to the pathophysiology of demyelinating disorders and nerve injury. PMID- 12898532 TI - Expression of 2',3'-cyclic nucleotide 3'-phosphodiesterase (CNPase) in the developing olfactory bulb and subventricular zone rostral extension. AB - The olfactory bulb (OB) presents a unique pattern of permanent acquisition of primary afferents and interneurons, but not much detail is known about the differentiation of its oligodendroglial cells. We studied the expression of 2',3' cyclic nucleotide 3'-phosphodiesterase (CNPase), a protein related to axonal ensheathment by myelinating cells. Expression of CNPase in OB follows a general caudorostral gradient, with the exception of the glomerular layer (GL). At postnatal day 5-6 (P5-P6), the first CNPase(+) profiles appeared in the dorsal lateral olfactory tract adjacent to the accessory OB (AOB), followed by rare cell bodies and processes in AOB internal plexiform layer at P7. At P9, the main OB (MOB) granular cell layer (GrCL) already showed intensely stained CNPase(+) processes. From P5 to P12, small numbers of CNPase(+) cells were found in the subventricular zone (SVZ), throughout its rostral extension (SVZ-RE), and in the intrabulbar subependymal layer. The appearance of CNPase(+) profiles delimiting glomeruli started in the GL rostralmost region at P12, extending to all GL levels, but glomeruli remained open caudally at P15. At P18, oligodendroglial glomeruli were evident throughout OB, but the adult pattern was established only after P30. There was no age-related loss of CNPase immunoreactivity in glial cell bodies, possibly indicating de novo ensheathment of neurites. Our results show an earlier onset of oligodendroglial differentiation in OB than previously reported and a rostrocaudal gradient of formation of oligodendroglial glomeruli. They also raise the possibility that a minor fraction of OB oligodendrocytes might derive from the SVZ-RE. PMID- 12898533 TI - Astrocyte-targeted expression of interleukin-6 protects the central nervous system during neuroglial degeneration induced by 6-aminonicotinamide. AB - 6-aminonicotinamide (6-AN) is a niacin antagonist, which leads to degeneration of gray matter astrocytes mainly in the brainstem. We have examined the role of interleukin-6 (IL-6) in this degenerative process by using transgenic mice with astrocyte-targeted IL-6 expression (GFAP-IL6 mice). This study demonstrates that transgenic IL-6 expression significantly increases the 6-AN-induced inflammatory response of reactive astrocytes, microglia/macrophages, and lymphocytes in the brainstem. Also, IL-6 induced significant increases in proinflammatory cytokines IL-1, IL-12, and tumor necrosis factor-alpha as well as growth factors basic fibroblast growth factor (bFGF), transforming growth factor-beta, neurotrophin-3, angiopoietin, vascular endothelial growth factor, and the receptor for bFGF. In accordance, angiogenesis was increased in GFAP-IL6 mice relative to controls after 6-AN. Moreover, oxidative stress and apoptotic cell death were significantly reduced by transgenic IL-6 expression. IL-6 is also a major inducer in the CNS of metallothionein I and II (MT-I+II), which were significantly increased in the GFAP-IL6 mice. MT-I+II are antioxidants and neuroregenerative factors in the CNS, so increased MT-I+II levels in GFAP-IL6 mice could contribute to the reduction of oxidative stress and cell death in these mice. PMID- 12898534 TI - Unusual topographical pattern of proximal astrogliosis around a cortical devascularizing lesion. AB - Class II vessels were disrupted on the cortical surface of adult rats within a circular 5-mm-diameter area. This consistently resulted in the formation of a conical lesion by day 1, with a cystic cavity forming by day 21. Four markers were used to identify the glial response surrounding the lesion. The antibody used against S100beta marked the largest astrocytic pool in the gray matter of the cerebral cortex; only approximately 5% of astrocytes were glial fibrillary acidic protein (GFAP)(+) in control animals. GFAP served as a marker for distal reactive gliosis and vimentin (VIM) for proximal gliosis. Isolectin B4 was used as an additional marker to distinguish VIM(+) microglia from astrocytes inside the lesion area. Three immunohistochemically distinct areas of reactive astrocytes surrounding the lesion were found within 24 hr of injury and lasted through day 6. The first area, in contrast to focal traumatic injuries, consisted of a 196-microm-thick boundary layer of S100beta(+) cells immediately surrounding the lesion that never expressed GFAP or VIM by day 6. This boundary layer turns into a GFAP(+) glial limitans encasing the cystic cavity by day 21. A second unusual extended area around the base of the lesion reaching partly into the corpus callosum consisted of S100beta(+)/GFAP(+)/VIM(+) cells. This region appears to be compatible with the local or proximal gliotic response usually found completely surrounding other focal-type injuries. The proximal response at the base of the lesion developed over the first 3 days in the following sequence: S100beta(+)/GFAP(-)/VIM(-) to S100beta(+)/GFAP(+)/VIM(-) to S100beta(+)/GFAP(+)/VIM(+). Ninety percent of the astrocytes in this area express VIM. This is very high compared with findings in stab-wound preparations, where only 10% of astrocytes (surrounding entire lesion) are found to be VIM(+). A third region, consistent with a remote or distal reactive gliotic response, demonstrated staining for S100beta and had increased GFAP contents throughout the neocortical hemisphere. Cells in this region were never found to be VIM(+). Among S100beta(+) cells close to the boundary region, more than 80% expressed detectable GFAP by 2 days after lesioning. S100beta(+) cells 1 mm more laterally (distal to lesion) did not express GFAP to the same level until day 6. Thus, we find three immunohistochemically distinct populations of reactive astrocytes surrounding the focal ischemic lesion. In contrast to the case for stab-wound traumatic injury, the response closest to and surrounding the lesion did not up regulate GFAP or VIM by day 6. The proximal response was, instead, more remote and only at the base of the lesion, extending partly into the corpus callosum. PMID- 12898535 TI - FGF-2 modulates expression and distribution of GAP-43 in frog retinal ganglion cells after optic nerve injury. AB - Basic fibroblast growth factor (bFGF or FGF-2) has been implicated as a trophic factor that promotes survival and neurite outgrowth of neurons. We found previously that application of FGF-2 to the proximal stump of the injured axon increases retinal ganglion cell (RGC) survival. We determine here the effect of FGF-2 on expression of the axonal growth-associated phosphoprotein (GAP)-43 in retinal ganglion cells and tectum of Rana pipiens during regeneration of the optic nerve. In control retinas, GAP-43 protein was found in the optic fiber layer and in optic nerve; mRNA levels were low. After axotomy, mRNA levels increased sevenfold and GAP-43 protein was significantly increased. GAP-43 was localized in retinal axons and in a subset of RGC cell bodies and dendrites. This upregulation of GAP-43 was sustained through the period in which retinal axons reconnect with their target in the tectum. FGF-2 application to the injured nerve, but not to the eyeball, increased GAP-43 mRNA in the retina but decreased GAP-43 protein levels and decreased the number of immunopositive cell bodies. In the tectum, no treatment affected GAP-43 mRNA but FGF-2 application to the axotomized optic nerve increased GAP-43 protein in regenerating retinal projections. We conclude that FGF-2 upregulates the synthesis and alters the distribution of the axonal growth-promoting protein GAP-43, suggesting that it may enhance axonal regrowth. PMID- 12898536 TI - Glucose uptake in PC12 cells: GLUT3 vesicle trafficking and fusion as revealed with a novel GLUT3-GFP fusion protein. AB - The distribution of glucose transporters at the cell surface has a major impact on cellular glucose uptake. In muscle cells and adipocytes, this distribution is under the control of insulin; however, neuronal glucose uptake is not acutely regulated by insulin. Factors that affect the translocation of the neuronal glucose transporter isoform GLUT3 vesicles to and their fusion with the plasma membrane are not well understood. We report that GLUT3 in PC12 cells colocalizes with SNARE complex proteins SNAP-25 and syntaxin 1, suggesting that fusion of GLUT3-containing vesicles with the plasma membrane is mediated by these proteins. In addition, it seems that GLUT3 vesicle fusion is regulated, as depolarization increases GLUT3 insertion into the plasma membrane. To study the dynamics of GLUT3 vesicle trafficking, we have created a GLUT3-GFP fusion protein that is easily expressed in PC12 cells. Trafficking of GLUT3-GFP seems normal, as 1). its distribution is similar to endogenous GLUT3, 2). GLUT3-GFP containing vesicles fuse with the plasma membrane evidenced by labeling of the fusion protein with an antibody directed against the exofacial epitope of GLUT3, and 3). glucose uptake is similar to PC12 cells not transfected with GLUT3 fusion protein. These studies are the first to examine GLUT3 trafficking and fusion in PC12 cells, and establish a model system to study regulation of the neuronal glucose transporter. PMID- 12898537 TI - Differential modulation of hippocampal chemical-induced injury response by ebselen, pentoxifylline, and TNFalpha-, IL-1alpha-, and IL-6-neutralizing antibodies. AB - The proinflammatory cytokines tumor necrosis factor (TNFalpha), interleukin-1 (IL 1alpha), and interleukin-6 (IL-6) have been associated with various models of hippocampal damage. To examine their role in initiation of an acute hippocampal injury response, 21-day-old male CD-1 mice received an acute intraperitoneal (i.p.) injection of trimethyltin hydroxide (TMT; 2.0 mg/kg) to produce necrosis of dentate granule neurons, astrocyte, and microglia reactivity. Tremors and intermittent seizures were evident at 24 hr. Intercellular adhesion molecule-1 (ICAM-1), glial fibrillary acidic protein (GFAP), anti-apoptotic TNFalpha inducible early response gene (A-20), macrophage inflammatory protein (MIP) 1alpha, TNFalpha, IL-1alpha, IL-6, and caspase 3 mRNA levels were significantly elevated. Pretreatment with the antioxidant, ebselen, decreased ICAM-1, A-20, and TNFbeta elevations. Pentoxifylline blocked elevations in A-20 and decreased elevations in GFAP mRNA levels. Neither prevented histopathology or behavioral effects. Intracisternal injection of TNFalpha-neutralizing antibody significantly inhibited both behavioral effects and histopathology. RNase protection assays showed that TMT-induced elevations in mRNA levels for ICAM-1, A-20, GFAP, MIP 1alpha, IL-1alpha, TNFalpha, TNFbeta, and caspase 3 were blocked by anti TNFalpha. These data demonstrate a significant role for TNFalpha in an acute neuro-injury in the absence of contribution from infiltrating cells. The cerebellum shows limited if any damage after TMT; however, in combination with the i.c.v. injection, elevations were seen in GFAP and in EB-22, a murine acute phase response gene homologous to the alpha (1)-antichymotrypsin gene. Elevations were similar for artificial cerebral spinal fluid and anti-IL-1alpha, and significantly increased with anti-TNFalpha, anti-IL-6, or the combination of antibodies. Responses seen in the cerebellum suggest synergistic interactions between the baseline state of the cell and manipulations in the cytokine environment. Data suggests a role for TNFalpha in the pathogenesis of hippocampal injury induced by TMT. PMID- 12898538 TI - Administration of the p38 MAPK inhibitor SB203580 affords brain protection with a wide therapeutic window against focal ischemic insult. AB - We have reported previously the delayed and differential induction of p38alpha and p38beta mitogen-activated protein kinases (MAPKs) in microglia and astrocytes, respectively, in brain after transient global ischemia. We report here the sustained induction and activation of p38alpha MAPK in activating microglia in rat brain after transient middle cerebral artery occlusion (MCAO). The intraventricular administration of SB203580, a p38 MAPK inhibitor, 30 min before MCAO reduced the infarct volume to 50% of the control, which was accompanied by the significant improvement of neurological deficits. More interestingly, the infarct volume was reduced to 72% and 77% when SB203580 was administered 6 hr and 12 hr after MCAO, respectively. The induction of various factors involved in inflammatory processes, such as inducible nitric oxide synthase (iNOS), tumor necrosis factor-alpha (TNF-alpha), interleukin-1beta (IL 1beta) and cyclooxygenase-2 (COX-2), was suppressed by the administration of SB203580 at 6 hr after MCAO. These results suggest that sustained activation of p38 MAPK pathway and p38 MAPK-associated inflammatory processes play a crucial role in postischemic brain. PMID- 12898539 TI - Spatiotemporal changes of apolipoprotein E immunoreactivity and apolipoprotein E mRNA expression after transient middle cerebral artery occlusion in rat brain. AB - Apolipoprotein E (ApoE) is a constituent of lipoprotein and plays an important role in the maintenance of neural networks. However, spatiotemporal differences in ApoE expression and its long-term role in neural process after brain ischemia have not been studied. We investigated changes of ApoE immunoreactivity and ApoE mRNA expression both in the core and in the periischemic area at 1, 7, 21, or 56 days after 90 min of transient middle cerebral artery occlusion. Double stainings for ApoE plus NeuN or plus ED1 were performed in order to identify cell type of ApoE-positive stainings. The maximal increase of ApoE expression was observed at 7 days in the core and at 7 and 21 days in the periischemic area. In the core, ApoE plus NeuN double-positive cells increased at 1 and 7 days, without ApoE mRNA expression, whereas they increased in the periischemic area, with a peak at 21 days, with ApoE mRNA expression in glial cells but not in neurons. On the other hand, ApoE plus ED1 double-positive cells increased only in the core, with a peak in number at 7 and 21 days and marked ApoE mRNA expression in macrophages. The present study suggests that ApoE plays various important roles in different type of cells, reflecting spatiotemporal dissociation between degenerative and regenerative processes after brain ischemia, and that ApoE is profoundly involved in pathological conditions, such as brain ischemia. PMID- 12898540 TI - Potential roles of Alzheimer precursor protein A4 and beta-amyloid in survival and function of aged spinal motor neurons after axonal injury. AB - To study the potential role of Alzheimer precursor protein A4 (APP) and beta amyloid (A/beta) on aging motor neuron survival, expression of APP, A/beta, and choline acetyltransferase (ChaT) were investigated in aged rats after either distal axotomy or root avulsion injury. Approximately 45% in number of total aged spinal motor neuron were normally APP-positive. A/beta-positive neurites were observed normally in the spinal ventral horn of aged rats. After distal axotomy, without apparent neurodegeneration such as cell loss and decreased ChaT immunoreactivity, increased levels of APP expression were observed in the spinal cords of aged rats post-injury. In contrast, after avulsion, expression of APP and A/beta were downregulated in the spinal ventral horn of aged rats, and marked loss of spinal motor neurons and downregulated ChaT expression were observed. Our data indicate that APP and A/beta might play beneficial roles in neuronal survival of aged spinal motor neurons after axonal injury. PMID- 12898541 TI - Hyperalgesic effects of gamma-aminobutyric acid transporter I in mice. AB - The present study focused on the involvement of gamma-aminobutyric acid transporter I (GAT1) in pain. We found that GABA uptake was increased in mouse spinal cord at 20 min and 120 min after formalin injection and in mouse brain at 120 min, but not 20 min, after formalin injection. In addition, the antinociceptive effects of GAT1-selective inhibitors were examined using assays of thermal (tail-flick) and chemical (formalin and acetic acid) nociception in C57BL/6J mice. The GAT1-selective inhibitors, ethyl nipecotate and NO-711, exhibited significant antinociceptive effects in these nociceptive assays. To study further the effects of GAT1 on pain, we used two kinds of GAT1 overexpressing transgenic mice (under the control of a CMV promoter or a NSE promoter) to examine the nociceptive responses in these mice. In the thermal, formalin, and acetic acid assays, both kinds of transgenic mice displayed significant hyperalgesia after nociceptive stimuli. In addition, the micro opioid receptor antagonist naloxone had no influence on nociceptive responses in wild type and transgenic mice. The results indicate that GAT1 is involved in the regulation of pain processes, and point to the possibility of developing analgesic drugs that target GAT1 other than opioid receptors. PMID- 12898543 TI - Adjusting power for a baseline covariate in linear models. AB - The analysis of covariance provides a common approach to adjusting for a baseline covariate in medical research. With Gaussian errors, adding random covariates does not change either the theory or the computations of general linear model data analysis. However, adding random covariates does change the theory and computation of power analysis. Many data analysts fail to fully account for this complication in planning a study. We present our results in five parts. (i) A review of published results helps document the importance of the problem and the limitations of available methods. (ii) A taxonomy for general linear multivariate models and hypotheses allows identifying a particular problem. (iii) We describe how random covariates introduce the need to consider quantiles and conditional values of power. (iv) We provide new exact and approximate methods for power analysis of a range of multivariate models with a Gaussian baseline covariate, for both small and large samples. The new results apply to the Hotelling-Lawley test and the four tests in the "univariate" approach to repeated measures (unadjusted, Huynh-Feldt, Geisser-Greenhouse, Box). The techniques allow rapid calculation and an interactive, graphical approach to sample size choice. (v) Calculating power for a clinical trial of a treatment for increasing bone density illustrates the new methods. We particularly recommend using quantile power with a new Satterthwaite-style approximation. PMID- 12898542 TI - Acquisition of an appetitive behavior prevents development of stress-induced neurochemical modifications in rat nucleus accumbens. AB - In rats, exposure to chronic unavoidable stress produces a decrease in dopamine output in the nucleus accumbens shell that is accompanied by a decreased density of the dopamine transporter and an increased activity of the dopamine-D(1) receptor complex. These modifications have been hypothesized to be adaptive to decreased dopamine output in stressed rats. We investigated whether the learning of an appetitive behavior sustained by palatable food, which is associated with increased dopamine output in the nucleus accumbens shell as measured by microdialysis experiments, would affect the modifications induced by chronic stress exposure on dopamine transporter density and dopamine-D(1) receptor complex activity in the nucleus accumbens. Rats exposed to chronic unavoidable stress after acquisition of the appetitive behavior showed a higher dopamine extraneuronal release in the nucleus accumbens shell than that of stressed animals, and similar to that of control rats. Moreover, previous acquisition of the appetitive behavior prevented development of a stress-induced decrease in dopamine transporter density, measured by [(3)H]-WIN 35428 binding, a stress induced increase in dopamine-D(1) receptor density, measured by binding of [(3)H] SCH 23390, and SKF 38393-stimulated adenylyl cyclase activity in the nucleus accumbens. These results support the hypothesis that changes induced in pre- and postsynaptic dopaminergic transmission by chronic stress exposure are related to decreased dopamine output. PMID- 12898544 TI - On the performance of random-coefficient pattern-mixture models for non-ignorable drop-out. AB - Random-coefficient pattern-mixture models (RCPMMs) have been proposed for longitudinal data when drop-out is thought to be non-ignorable. An RCPMM is a random-effects model with summaries of drop-out time included among the regressors. The basis of every RCPMM is extrapolation. We review RCPMMs, describe various extrapolation strategies, and show how analyses may be simplified through multiple imputation. Using simulated and real data, we show that alternative RCPMMs that fit equally well may lead to very different estimates for parameters of interest. We also show that minor model misspecification can introduce biases that are quite large relative to standard errors, even in fairly small samples. For many scientific applications, where the form of the population model and nature of the drop-out are unknown, interval estimates from any single RCPMM may suffer from undercoverage because uncertainty about model specification is not taken into account. PMID- 12898546 TI - Separation of individual-level and cluster-level covariate effects in regression analysis of correlated data. AB - The focus of this paper is regression analysis of clustered data. Although the presence of intracluster correlation (the tendency for items within a cluster to respond alike) is typically viewed as an obstacle to good inference, the complex structure of clustered data offers significant analytic advantages over independent data. One key advantage is the ability to separate effects at the individual (or item-specific) level and the group (or cluster-specific) level. We review different approaches for the separation of individual-level and cluster level effects on response, their appropriate interpretation and give recommendations for model fitting based on the intent of the data analyst. Unlike many earlier papers on this topic, we place particular emphasis on the interpretation of the cluster-level covariate effect. The main ideas of the paper are highlighted in an analysis of the relationship between birth weight and IQ using sibling data from a large birth cohort study. PMID- 12898545 TI - Error in timing in regression with observed longitudinal measurements. AB - We consider regression analysis of a disease outcome in relation to longitudinal data which are observations from a random effects model. The covariate variables of interest are the values of the underlying trajectory at some time points, which may be fixed or subject-specific. Because the underlying random coefficients are unknown, the covariates to the primary model are generally unobserved. In addition, measurements are often not observed at the time points of interest. A motivating example to our model is the effects of age at adiposity rebound and the associated body mass index on the risk of adult obesity. The adiposity rebound is a time point at which the trajectory of a child's body fatness declines to a minimum. This general error in timing problem may be applied to an analysis when time-dependent marker variables follow a polynomial model in which the effect of a local maximum or minimum point may be of interest. It can be seen that directly applying estimated covariates, possibly obtained from estimated time points, may lead to bias. Estimation procedures based on expected estimating equations, regression calibration and simulation extrapolation are applied to this problem. PMID- 12898547 TI - Two bivariate geometrically defined reference regions with applications to male reproductive hormones and human growth. AB - Two conceptually simple two-dimensional reference charts are described. The charts are characterized analytically, but also by geometric properties that are easily communicated to subject-matter researchers--the users of such charts. For both charts, the proposed estimators are non-parametric, so no distributional assumptions are needed about the two-dimensional response. An important example that will serve to motivate and illustrate one of the charts is the study of the human reproductive hormones Inhibin B and FSH. A low Inhibin B concentration combined with a high FSH concentration is an indication that something may be wrong in the seminal cells which produce semen, and such measurements help in the process of distinguishing between different reasons for infertility. The aim in the study of the human reproductive hormones was to give a two-dimensional reference curve for the non-normally distributed hormone measurements while taking into account that some measurements were left-censored because of a detection limit in the assay used to measure the concentrations. PMID- 12898548 TI - Estimation of k for the poly-k test with application to animal carcinogenicity studies. AB - This paper extends the survival-adjusted Cochran-Armitage test in order to achieve improved robustness to a variety of tumour onset distributions. The Cochran-Armitage test is routinely applied for detecting a linear trend in the incidence of a tumour of interest across dose groups. To improve the robustness to the effects of differential mortality across groups, Bailer and Portier introduced the poly-3 test by a survival adjustment using a fractional weighting scheme for subjects not at full risk of tumour development. The performance of the poly-3 test depends on how closely it represents the correct specification of the time-at-risk weight in the data. Bailer and Portier further suggested that this test can be improved by using a general k reflecting the shape of the tumour onset distribution. In this paper, we propose a method to estimate k by equating the empirical lifetime tumour incidence rate obtained from the data based on the fractional weighting scheme to a separately estimated cumulative lifetime tumour incidence rate. This poly-k test with the statistically estimated k appears to perform better than the poly-3 test which is conducted without prior knowledge of the tumour onset distribution. Our simulation shows that the proposed method improves the robustness to various tumour onset distributions in addition to the robustness to the effects of mortality achieved by the poly-3 test. Large sample properties are shown via simulations to illustrate the consistency of the proposed method. The proposed methods are applied to analyse two real data sets. One is to find a dose-related linear trend on animal carcinogenicity, and the other is to test an effect of calorie restriction on experimental animals. PMID- 12898549 TI - Characterizing classes of antiretroviral drugs by genotype. AB - This paper develops methods for using HIV-1 genotypic information to group patients who are expected to have similar patterns of sensitivity or resistance to two or more drugs. The methods presented are an extension of prediction based classification to handle multiple drug responses. Here, the goal is to determine the probability that one antiretroviral therapy will be more favourable than another for an individual given the specific genotypic or other characteristics of the infecting viral population. This approach requires a model relating genotype to a vector of drug specific phenotypic responses. A comparison of Nelfinavir and Indinavir is provided using 2746 protease sequences and corresponding in vitro sensitivity assays provided to us by the Virco Group. PMID- 12898550 TI - Use of max and min scores for trend tests for association when the genetic model is unknown. AB - In case-control studies, the Cochran-Armitage (CA) trend test is powerful for detection of an association between a risk allele and a marker. To apply this test, a score should be assigned to the genotypes based on the genetic model. When the underlying genetic model is unknown, the trend test statistic is a function of the score. In this paper, simple procedures are given to obtain two scores (max and min), which respectively maximize and minimize the CA trend test statistics for genetic associations. These two scores can be used to examine the effect of the choice of scores on the test of no association. When the CA trend test statistic with the max (or min) score is less (or greater) than a prespecified value, the conclusion is clear: we will accept (or reject) the null hypothesis of no association for any scores used. When this value is less than the CA trend test statistic with the max score but greater than the one with the min score, the decision of whether or not to reject the null hypothesis depends on the choice of scores. In this situation, the CA trend test with a prespecified score cannot be used without careful scientific justification of the choice of scores. The use of max and min scoring schemes is applied to a real data set. PMID- 12898551 TI - A simple confidence interval for meta-analysis. K. Sidik and J. N. Jonkman, Statistics in Medicine 2002; 21:3153-3159. PMID- 12898554 TI - The role of palatable food and hunger as trigger factors in an animal model of stress induced binge eating. AB - OBJECTIVE: Dieting and stress are etiological factors in eating disorders, and dieting strongly predicts stress-induced overeating in the nonclinical population. We developed an animal model of binge eating in sated rats that is evoked by stress, but only in rats with a history of caloric restriction and only if highly palatable food (HPF) is available after stress. This study investigated the effect of known binge triggers, a taste of HPF and of hunger, on this type of binge eating. METHOD: Female rats were cycled through the R/S protocol but this time were given just a taste of HPF with ad lib regular chow. After another R/S cycle, rats were stressed during restriction (while hungry) and were given HPF and chow. RESULTS: Although binge eating did not occur if only chow was available after stress, just a taste of HPF sufficed to increase chow intake to more than 160% (p < 0.001) of rats with a history of restriction only, stress-only, or neither. Hunger increased the proportion of chow consumed by both restricted groups, but stress magnified this hunger-induced overeating by increasing HPF intake to 137% of restriction-only rats (p < 0.001). DISCUSSION: These effects suggest that binge eating in this model is motivated by reward, not metabolic need, and parallels observations of binge triggers described in clinical binge eating disorders. This strengthens the validity of using this animal model to target the physiology and treatment of eating disorders preceded by dieting and stress. PMID- 12898555 TI - A new animal model of binge eating. PMID- 12898556 TI - Requiring remission of undue influence of weight and shape on self-evaluation in the definition of recovery for bulimia nervosa. AB - OBJECTIVE: The current study evaluated the concurrent validity of requiring remission of undue influence of weight and shape on self-evaluation (undue influence) in defining recovery from bulimia nervosa (BN). METHODS: Three groups completed the Beck Depression Inventory, the Mood and Anxiety Symptom Questionnaire, the Body Shape Questionnaire, and the Social Adjustment Scale: 31 women were fully recovered from BN (FR), 28 women had no behavioral symptoms of BN (partially recovered [PR]), and 59 matched non-eating-disordered controls (MC). RESULTS: The PR group had more pathologic scores on depression, anxiety, body dissatisfaction, and social adjustment compared with both the FR and MC groups, which did not differ from each other. DISCUSSION: These findings suggest that including remission of cognitive symptoms in a standardized definition of recovery may prove to be clinically useful in establishing reliable prognostic indicators. Future research should evaluate the role played by cognitive symptoms in triggering relapse. PMID- 12898557 TI - Anxiety and psychoactive substance use disorder comorbidity in anorexia nervosa or depression. AB - OBJECTIVE: This study considered whether the prevalence and type of anxiety and psychoactive substance use disorder (PSUD) diagnoses differ between women with spectrum anorexia nervosa (AN) (N=40) and women with major depressive disorder (N = 58) participating in outpatient clinical trials. METHOD: Anxiety and PSUD diagnoses (according to criteria in the 3rd Rev. ed. of the Diagnostic and Statistical Manual of Mental Disorders) were assessed using structured clinical interviews. Comparisons were made between AN subtypes (restricting or binge eating/purging) and by history of depression within the AN sample. RESULTS: A high prevalence of obsessive-compulsive disorder (OCD) was found in women with AN. However, social phobia, simple phobia, and PSUD were significantly elevated in both women with depression and women with AN. Prevalences were similar for anxiety and PSUD diagnoses between AN subtypes. DISCUSSION: Women with anorexia or depression were comparable in all respects, except for the elevated OCD prevalence in AN, emphasizing the need to use clinical comparison groups to avoid inadvertently attributing elevated prevalences of comorbid conditions to specific disorders. PMID- 12898558 TI - Stress situation reveals an association between perfectionism and drive for thinness. AB - OBJECTIVE: Several theorists have hypothesized that stress situations may trigger abnormal eating and even eating disorders in individuals with a perfectionistic personality. The purpose of this study was to assess whether a stress situation would reveal an association between perfectionism and measures of eating disorders among female high school students. METHODS: A sample of 42 female high school students completed the Multidimensional Perfectionism Scale and the Eating Disorder Inventory three times: on an average school day, on the day of an examination, and on the day they received the evaluation of the examination. Linear regression analysis was calculated to verify whether the dimensions of perfectionism were associated with the measures of eating disorders. RESULTS: Body Dissatisfaction was associated with perfectionism on all the three occasions, whereas Drive for Thinness was associated with perfectionism only on the day the students received the results of the examination. DISCUSSION: The results suggest that among nonclinical female individuals, stress may make significant a previously absent association between perfectionism and an actual desire or plan to lose weight. Such a finding suggests that stress may stimulate behaviors related to eating disorders in individuals with a perfectionistic personality. PMID- 12898559 TI - Cue exposure in the treatment of resistant bulimia nervosa. AB - OBJECTIVE: It was hypothesized that binge eating (bulimia nervosa [BN]) may be caused by the anticipatory and immediate anxiety associated with certain types of food. Consequently, an extinction schedule should reduce binge eating. METHODS: Cue exposure was carried out with 6 bulimic women who had responded poorly or not at all to the usual pharmacologic or cognitive-behavioral treatments. RESULTS: Binge eating and vomiting were almost totally suppressed in the 6 patients. Symptom suppression was maintained at two follow-ups, one at 4-20 months and another at 2.5-3 years. DISCUSSION: Cue exposure may be effective with BN that is resistant to conventional treatments. The anxiety associated with food plays an important role in provoking and/or maintaining binge eating. Motivation to change is likely to be an important mediator. PMID- 12898560 TI - Binge eating and psychopathology in severely obese adolescents. AB - OBJECTIVE: The authors describe binge eating and psychopathology in severely obese adolescents who are seeking treatment for obesity and search for specific psychological features that may be associated with binge eating. METHOD: Obese adolescents (n = 102) were assessed with the Binge Eating Scale (BES), the State Trait Anxiety Inventory for Children (STAIC), the Beck Depression Inventory (BDI), Coopersmith's Self-Esteem Inventory (SEI), and the Body-Esteem subscale from the Piers-Harris Children's Self-Concept Scale (PHCSS). They were also evaluated with the Montgomery and Asberg Depression Rating Scale (MADRS) and the Brief Scale for Anxiety (BSA). Two subgroups (i.e., binge eaters and non-binge eaters) were then established according to the BES score. RESULTS: Binge eating symptoms were frequent in this population and the binge eating dimension was related to high levels of anxiety and depression, as well as to low levels of self-esteem and body-esteem. The dimensions of anxiety and depression were associated specifically with binge eaters. DISCUSSION: As observed in the obese adult population, binge eating symptoms were found frequently in severely obese adolescents seeking treatment and were related strongly to studied parameters. Obese adolescents who binge eat are a subgroup with high psychopathologic distress. PMID- 12898561 TI - Body images of 4-8-year-old girls at the outset of their first artistic gymnastics class. AB - OBJECTIVE: The primary objective of this investigation was to compare body images of 54 girls (4-8 years old) who were beginning their first gymnastics class to 54 age-matched (+/-1 year) and % body fat-matched (+/-3%) girls not participating in gymnastics. METHOD: Ratings of actual and ideal body size and a computed measure of body dissatisfaction (actual minus ideal) were obtained. Body composition was measured with dual-energy x-ray absorptiometry. RESULTS: Girls beginning gymnastics did not differ significantly from controls on (mean +/- SD) actual (2.2 +/- 1.4 vs. 2.1 +/- 1.4), ideal (1.8 +/- 1.3 vs. 1.8 +/- 1.5), or body dissatisfaction (0.39 +/- 1.8 vs. 0.26 +/- 1.4) scores. DISCUSSION: Body images of 4-8-year-old girls beginning their first artistic gymnastics class do not differ from non-gymnast controls matched on age and % body fat. This observation casts doubt on the idea that young girls who are dissatisfied with their body and want to be smaller are more likely to enroll in gymnastics classes than girls without this type of body dissatisfaction. PMID- 12898562 TI - Are adolescents harmed when asked risky weight control behavior and attitude questions? Implications for consent procedures. AB - OBJECTIVES: This study explores whether asking minors about risky weight control behaviors and attitudes increases the frequency of those behaviors and attitudes. METHODS: Participants were 115 sixth-grade girls who responded to questions on risky weight control behaviors and attitudes at baseline and at 12-month follow up. An additional 107 girls, who had not been part of the baseline, provided data only at follow-up. The two groups were compared on risky weight control behaviors and attitudes at follow-up using chi-square analyses, Mann-Whitney U tests, Cohen's effect sizes, and odds ratios. RESULTS: No evidence of a negative effect in the twice-assessed group was found. All rates decreased from baseline to follow-up. CONCLUSIONS: There is only minimal risk and perhaps even some benefit of asking questions about risky weight control behaviors and attitudes. Implications for determining appropriate consent procedures are discussed. PMID- 12898563 TI - Investigating the structure of the eating inventory (three-factor eating questionnaire): a confirmatory approach. AB - OBJECTIVE: To investigate the internal structure of the Eating Inventory using confirmatory factor analytic (CFA) methods. METHOD: Female participants in a population-based twin study (N = 1,510) completed a reduced version of the Eating Inventory as part of a larger study of eating behaviors. Two CFA estimation methods were implemented using LISREL and Mplus to evaluate three different factor structures reported in the literature. RESULTS: Overall results for Stunkard and Messick's (1985) three-factor model indicated a poor fit to these data. The structure reported by Ganley resulted in an improper solution with factor correlations over 1 and several loadings estimated near 0 or with negative signs. The model proposed by Hyland et al. also fit poorly for all three models, the pattern of loadings on the Restraint factor proved most difficult to interpret. DISCUSSION: This study represents the first application of these confirmatory methods to the three subscales of the Eating Inventory. The instrument, as administered in this study, did not conform well to the structures proposed in previous studies. Because the Eating Inventory is recommended as a treatment planning tool, poor replication of the constructs purported to be assessed by this instrument underscores the importance of further internal validation research. PMID- 12898564 TI - Metoclopramide and unintended weight gain. AB - OBJECTIVE: This paper describes the case of a 19-year-old female with anorexia nervosa (restricting type) who was prescribed metoclopramide (Reglan, Robins, Richmond, VA) for delayed gastric emptying. She subsequently experienced dramatic and unintended weight gain. RESULTS: The initial treatment of delayed gastric emptying with metoclopramide did not result in a significant change in body weight. However, an increase in the dose resulted in a 20% increase in the patient's body weight over a 2-month period. DISCUSSION: The risks of weight gain that accompany the administration of metoclopramide are discussed, as well as the postulated etiology and psychological implications for the patient. PMID- 12898565 TI - Reduced febrile response to bacterial infection in anorexia nervosa patients. AB - OBJECTIVE: To report a reduced febrile response to bacterial infections in anorexia nervosa (AN) patients. METHOD: Four cases were obtained from a retrospective review of charts from the St. Paul's Hospital Eating Disorders Program (Vancouver, Canada). The patients had died or had been admitted to the hospital for treatment of a bacterial infection. In addition, one case was obtained from the Royal Prince Alfred Hospital (Sydney, Australia). RESULTS: All patients suffered a bacterial infection during the course of AN. None of the patients had a temperature higher than 37 degrees C during the infectious illness. DISCUSSION: The absence of fever in AN may delay the diagnosis of bacterial infection and may be a marker of an impaired immune response. Therefore, alternative methods of investigation are necessary in patients with AN suspected of having a bacterial infection. PMID- 12898566 TI - A characteristic reactive hypoglycemia induced by rapid change of eating behavior in anorexia nervosa: a case report. AB - The relationship among reactive hypoglycemia, corresponding insulin metabolism, and eating behavior in anorexia nervosa (AN) has not been well documented. METHOD: A 22-year-old woman with AN was admitted to Kagoshima University Hospital. An oral glucose tolerance test was performed with additional sampling at 15 minutes on the seventh hospital day when her daily caloric intake rapidly increased, reaching 2,000kcal perday. RESULTS: An elevated level of insulin secretion (1,190pmol/L) was observed during the very early phase (15 minutes) after glucose load, whereas corresponding blood glucose elevations were not as high (1.2mmol/L). The patient experienced asymptomatic hypoglycemia (2.1mmol/L). CONCLUSIONS: Our findings suggested that a rapid increase of energy intake might cause reactive hypoglycemia with characteristic insulin metabolism, that is, an elevated level of insulin secretion during the very early phase. PMID- 12898567 TI - Tumor necrosis factor haplotype analysis amongst schizophrenia probands from four distinct populations in the Asia-Pacific region. AB - A single nucleotide polymorphism (TNF(-308A)) within the promoter region of the gene encoding tumor necrosis factor (TNF), has been significantly associated with schizophrenia in a study of Italian patients and control subjects Boin et al. [2001: Mol Psychiatry 6:79-82]. We have applied case-control analyses to examine TNF promoter haplotypes (containing TNF(-308) and two additional promoter variants: TNF(-376) and TNF(-238)) in four schizophrenia cohorts drawn from Australian, Indian Fijian, Indigenous Fijian, and Brahmin populations. In addition, we have applied the sibling transmission disequilibrium (STD) test to promoter haplotypes within 81 trios drawn from Australian Caucasian pedigrees with multiple schizophrenia cases, and 86 trios drawn from the Brahmin population of Tamil Nadu province in Southern India. Within each of these cohorts, we found no evidence of recombination between these tightly linked promoter variants, supporting previous studies which demonstrated that only a subset of the eight possible haplotypes exist. Of the four observed haplotypes, we and others have observed only one carries the TNF(-308A) variant allele. We report no significant differences in TNF promoter haplotype frequencies between the patient and control groups within each population, although the Indian Fijian cohort showed a trend towards reduced TNF(-308A) alleles amongst schizophrenia cases (P = 0.07). We found no evidence of bias in TNF promoter haplotype transmission to schizophrenia probands. Very similar results were obtained when only the TNF(-308) polymorphism was considered. Taken together, these data provide no support for the involvement of TNF promoter variants TNF(-308), TNF(-376), and TNF(-238) in schizophrenia susceptibility within four ethnically distinct cohorts. PMID- 12898568 TI - Association of a haplotype in the serotonin 5-HT4 receptor gene (HTR4) with Japanese schizophrenia. AB - The serotonin 5-HT(4) receptor (5-HT(4)) is implicated in cognitive function, of which impairment is hypothesized as one of the core disturbances of schizophrenia. Linkage analysis shows that 5q33.2, in which HTR4 is located, is schizophrenia-susceptibility loci. We therefore hypothesized that variation in the 5-HT(4) receptor gene (HTR4) modifies genetic susceptibility to schizophrenia. HTR4 coding regions and introns that include the branch sites of HTR4 were investigated in 96 unrelated Japanese schizophrenics using denaturing high-performance liquid chromatography analysis. One silent single nucleotide polymorphism (SNP) within the coding region and six intronic SNPs were detected. 353 + 6G > A was located in the branch site that could be effect to RNA splicing. None of the four SNPs, in which rare-allele frequencies were more than 10% was associated with 189 schizophrenics, in comparison to 299 controls. However, a highly significant association between schizophrenia and haplotype A-T (OR = 0.13 [0.03-0.58]) was detected. These findings suggest that haplotype A-T itself may inhibit the occurrence of schizophrenia, or that another susceptible genetic variants may exist within linkage disequilibrium. PMID- 12898569 TI - CAG-repeat length in exon 1 of KCNN3 does not influence risk for schizophrenia or bipolar disorder: a meta-analysis of association studies. AB - Schizophrenia and bipolar disorder both show some evidence for genetic anticipation. In addition, significant expansion of anonymous CAG repeats throughout the genome has been detected in both of these disorders. The gene KCNN3, which codes for a small/intermediate conductance, calcium-regulated potassium channel, contains a highly polymorphic CAG-repeat array in exon 1. Initial evidence for association of both schizophrenia and bipolar disorder with increased CAG-repeat length of KCNN3 has not been consistently replicated. In the present study, we performed several meta-analyses to evaluate the pooled evidence for association with CAG-repeat length of KCNN3 derived from case-control and family-based studies of both disorders. Each group of studies was analyzed under two models, including a test for direct association with repeat length, and a test for association with dichotomized repeat-length groups. No evidence for a linear relationship between disease risk and repeat length was observed, as all pooled odds ratios approximated 1.0. Results of dichotomized allele-group analyses were more variable, especially for schizophrenia, where case-control studies found a significant association with longer repeats but family-based studies implicated shorter alleles. The results of these meta-analyses demonstrate that the risks for both schizophrenia and bipolar disorder are largely, if not entirely, independent of CAG-repeat length in exon 1 of KCNN3. This study cannot exclude the possibility that some aspect of this polymorphism, such as repeat-length disparity in heterozygotes, influences risk for these disorders. Further, it remains unknown if this polymorphism, or one in linkage disequilibrium with it, contributes to some distinct feature of the disorder, such as symptom severity or anticipation. PMID- 12898570 TI - Genome scan of the fifty-six bipolar pedigrees from the NIMH genetics initiative replication sample: chromosomes 4, 7, 9, 18, 19, 20, and 21. AB - The NIMH genetics initiative on bipolar disorder was established to collect uniformly ascertained bipolar pedigrees for genetic studies. In 1997, the four participating sites published a genome scan on the initial set of 97 bipolar pedigrees. Fifty-six additional bipolar pedigrees have now been ascertained and evaluated. This replication pedigree set contains 354 genotyped subjects, including 139 bipolar I (BPI) subjects, five schizoaffective bipolar type SA/BP subjects, 41 bipolar II (BPII) subjects, and 43 recurrent unipolar (RUP) depression subjects. Our site has recently genotyped the replication study bipolar pedigrees using 107 microsatellite markers from chromosomes 4, 7, 9, 18, 19, 20, and 21. We are now reporting parametric and nonparametric linkage results from this effort. Multipoint nonparametric linkage analysis produced three candidate regions with allele sharing LOD scores >/= 1.0. The linkage signal on 4q35 peaked between markers D4S3335 and D4S2390 with an allele sharing LOD score of 2.49. This finding exceeds standard criteria for suggestive linkage. Two additional loci approach suggestive linkage levels: the 4q32 finding had its maximum near marker D4S1629 with an allele sharing LOD score of 2.16, and the 20p12 finding peaked at D20S162 with an allele sharing LOD score of 1.82. Multipoint parametric linkage analysis produced similar findings. When we combined the genotype data from the original and the replication pedigree sets, 20p12 yielded a nonparametric LOD score of 2.38, which exceeds standard criteria for suggestive linkage, and a corresponding parametric HLOD score of 2.98. The combined analysis did not provide further support for linkage to 4q32 and 4q35. PMID- 12898571 TI - Linkage and association between serotonin 2A receptor gene polymorphisms and bipolar I disorder. AB - Several inconsistent associations between bipolar I disorder (BD1) and polymorphisms of the genes encoding the serotonin 2A receptor (HTR2A) have been published. We conducted the Transmission Disequilibrium Test (TDT) and case control comparisons involving nine single nucleotide polymorphisms at the serotonin 2A receptor gene (four SNPs of HTR2A exons and five flanking SNPs). Comparison of BD1 cases (n = 93) with a group of unrelated population based controls (n = 92) revealed associations with SNPs on exons 2 and 3 (516C/T and 1354C/T, respectively), consistent with haplotype-based differences. Analysis of the cases and their available parents using the TDT suggested significant linkage and associations with 1354C/T, as well as haplotypes bearing this SNP. Our results support an etiological role for HTR2A in BD1. In view of the relatively small sample, replicate studies using large samples are needed. PMID- 12898572 TI - Genetic dissection of psychopathological symptoms: insomnia in mood disorders and CLOCK gene polymorphism. AB - We investigated the possible effect of the 3111T/C CLOCK gene polymorphism on sleep disorders in a sample of 620 patients affected by major depressive disorder (MDD) and bipolar disorder (BP). We detected a significantly higher recurrence of initial (P = 0.0001), middle (P = 0.0009), and early (P = 0.0008) insomnia in homozygotes for the C variant and a similar trend concerning decreased need of sleep in BP (P = 0.0074). Other demographic and clinical features were found not related with CLOCK polymorphisms. This preliminary observation leads to hypothesize a possible involvement of the CLOCK gene polymorphism in the sleep disregulations in MDD and BP. PMID- 12898573 TI - The association of the D2S2944 124 bp allele with recurrent early onset major depressive disorder in women. AB - Major depressive disorder (MDD) and substance use disorders (SUD) are complex behavioral disorders with 40-50% heritability. Recently, Zubenko and colleagues reported that the 124 bp allele of D2S2944, a tetranucleotide repeat marker on 2q35, is strongly associated with recurrent, early onset MDD (RE-MDD) and alcohol use disorders in women. To test this hypothesis, we examined the association of the 124 bp allele in a subset of 171 adoptees from the Iowa Adoption Studies, a population with high rates of MDD and SUD. We report that in our population, the 124 bp allele significantly associated with RE-MDD in women. There was slight evidence of an increased of SUD in women with the 124 bp allele with the rate of cannabis use disorders reaching statistical significance (P < 0.04) before correction for multiple comparisons. Given the history of prior studies implicating 2q35 as a locus encoding vulnerability to co-morbid alcoholism and depression, these findings strongly suggest that sequence variation conveying increased susceptibility to MDD and possibly SUD is in close proximity to D2S2944. PMID- 12898574 TI - D2 and D4 dopamine receptor gene polymorphisms and personality traits in a young Korean population. AB - The correlation between the D4 dopamine receptor gene (DRD4) and the D2 dopamine receptor gene (DRD2) polymorphisms was investigated with personality traits. For this study, homogeneous population consisting of 243 young alcohol- and drug naive Koreans who were blood-unrelated with a mean age (+/-SD) of 13.87 (+/-0.30) years old was analyzed for the DRD4 and the DRD2 polymorphisms with their personality trait by Temperament and character inventory (TCI). The association between Novelty seeking (NS) score and DRD4 long alleles was only observed among the female subjects (t = 2.11, P = 0.037), but not in the male counter part. Female subjects who carried the DRD2 less frequent alleles (TaqI A1, TaqI B1, and Intron6 1) showed higher RD4 scores (dependence vs. independence) of Reward dependence (RD) than those without these alleles (P < 0.05). There was no interaction between DRD4 and DRD2 on the personality traits. These results, thus, confirmed the previous findings in which the long repeats of the DRD4-exon III polymorphism are related to NS personality trait, and also suggested that the DRD2 less frequent alleles were also associated with the reward-dependent trait. PMID- 12898576 TI - Report from the 4th international meeting of the attention deficit hyperactivity disorder molecular genetics network. AB - Investigators from around the world have convened annually since 1999 to share their work on the molecular genetic basis of attention deficit hyperactivity disorder (ADHD). The 4th meeting of the ADHD Molecular Genetics Network was held in May 2002. This conference began with a discussion of ongoing and proposed collaborative projects. Ongoing collaborations include the International Multi Center ADHD Genetics (IMAGE) project, a study of additive and interactive effects of DAT1 and DRD4 on ADHD and a meta-analysis of association studies of ADHD and the DRD5 gene. A new collaborative project regarding DNA pooling was also proposed. Over 20 investigators presented recent findings from their individual research sites. Participants also broke up into working groups to discuss additional projects and topics of mutual interest, including population genetic studies using dimensional phenotypic data and a QTL approach, endophenotypes for ADHD and the genetics of ADHD in adults. Finally, participant feedback was obtained about the utility of the ADHD Molecular Genetics Network conferences and the e-mail network. Although technological and statistical advances in the field of molecular genetics have allowed researchers to begin to identify specific genes that are associated with ADHD, continued collaborative efforts are needed to fully elucidate the genetic underpinnings of this complex phenotype. Data collected at the current conference indicate that the ADHD Molecular Genetics Network has facilitated new collaborative as well as single-site projects among ADHD researchers. PMID- 12898575 TI - Association of the 480 bp DAT1 allele with methylphenidate response in a sample of Irish children with ADHD. AB - Several studies have implicated the dopamine transporter gene (DAT1) as conferring susceptibility to attention deficit hyperactivity disorder (ADHD), in particular, a VNTR situated at the 3' end of the gene. In addition, the 10-repeat VNTR allele associated with ADHD has been reported to be associated with an over active transporter protein (DAT). Thus children possessing this variant might be particularly responsive to methylphenidate, a drug known to act by blocking DAT. We have examined this hypothesis and now report an association between the 10 repeat VNTR DAT1 polymorphism and retrospectively rated methylphenidate response in a sample of 119 Irish children with ADHD (chi(2) = 7.918, df = 1, P = 0.005). Our findings suggest a role for the 10-repeat DAT1 risk allele in medication response and may help to predict positive clinical outcome in ADHD. PMID- 12898577 TI - Segregation analysis of phenotypic components of learning disabilities. II. Phonological decoding. AB - Dyslexia is a common, complex disorder, which is thought to have a genetic component. The study of the genetics of dyslexia is complicated by a lack of consensus on diagnostic criteria, and the probability of genetic heterogeneity-it is possible that deficits in different language processes are caused by different underlying genes. In order to address these difficulties, we study continuous phenotypes that are part of the psychometric test batteries often used to diagnose dyslexia. Prior to embarking on a linkage study, it is helpful to employ segregation analysis, both to identify phenotypes that may be amenable to mapping by linkage analysis, and to determine the best models to use for model based analyses. We study 409 people in 102 nuclear families, and employ (1) oligogenic segregation analysis to estimate the number of quantitative trait loci (QTLs) contributing to each phenotype, and (2) complex segregation analysis in order to identify the most parsimonious inheritance model. In this paper, we consider two measures of phonological decoding ability-word attack and phonemic decoding efficiency. We find evidence for one or two genes of at least modest effect contributing to phonemic decoding efficiency, and the best fitting model is a dominant major gene model with residual familial correlations. For word attack, we find evidence for one or two genes of at least modest effect, and the variation in the trait is best explained by a polygenic model. PMID- 12898578 TI - A psychometrically derived impulsive trait related to a polymorphism in the serotonin transporter gene-linked polymorphic region (5-HTTLPR) in a Japanese nonclinical population: assessment by the Barratt impulsiveness scale (BIS). AB - Although a number of studies have shown that human impulsive traits are associated with indices of central serotonin function, few researchers have investigated the relationship between a polymorphism in the serotonin transporter gene-linked region (5-HTTLPR) and a psychometrically derived impulsive trait. We determined the 5-HTTLPR polymorphism in 123 employed Japanese male adults using the polymerase chain reaction. The distribution of allelic frequency was determined and also investigated the relationship of the 5-HTTLPR polymorphism to a impulsive trait as measured by the Barratt Impulsiveness Scale, 11th version (BIS-11). The distribution of allelic frequency was found to be almost identical to that previously reported in Japanese (the frequency for the long (L)/L, L/short (S), and S/S genotypes was: 3, 28, and 68%, respectively). In a comparison between the genotype groups, the S/S genotype group significantly higher scored for the total BIS-11 and the subscale attentional impulsiveness than the L/S + L/L genotype group. These findings suggest that individuals with a homozygous S-allele may be more impulsive than those with the other genotype. PMID- 12898579 TI - Association between human mu-opioid receptor gene polymorphism, pain tolerance, and opioid addiction. AB - Central to both pain responses and opioid addiction is activity at the micro opioid receptor. To explore the role of the micro -opioid receptor gene (OPRM) in human pain tolerance and opioid addiction, we examined the relationships among OPRM genotype and experimental pain tolerance in opioid addicts in methadone treatment (n = 50) and healthy normal controls (n = 59). Pain phenotype (pain tolerant vs. pain intolerant) was operationalized as tolerance to a standardized noxious stimulus (either thermal or mechanical), and dichotomized based on distribution. One microsatellite and two single nucleotide polymorphisms, A118G and C17T, in exon 1 were typed to study the OPRM gene. Although the established relationship between the phenotypes of opioid addiction and pain intolerance was validated (P = 0.02), genotype differed neither between addict-affected vs. control, nor pain tolerant vs. intolerant subjects. The variant A118G was absent in all individuals and the C17T polymorphism appeared in only three African American individuals (two addicts and one control). The absence of this polymorphism, the small sample size and the heterogeneous ethnic backgrounds of participants in the pilot study allow only tentative conclusions based on the results, thus the role of the opioid receptor in pain and opioid reward response remains uncertain. PMID- 12898580 TI - Linkage disequilibrium and haplotype analysis between serotonin receptor 1B gene variations and subtypes of alcoholism. AB - A number of studies have reported a possible association between serotonergic pathway genes and alcoholism. A silent polymorphism (G to C substitution) in the gene encoding the autoreceptor 5-HT1B was linked to antisocial alcoholism in Finnish and an American Indian populations [Lappalainen et al., 1998: Arch Gen Psychiatry 55:989-994]. Several other polymorphisms of this gene have been investigated for their association with neuropsychiatric disorders. In the present study, a sample of 133 alcoholics without and 39 alcoholics with medical complications, and 88 normal controls was screened for three single nucleotide polymorphisms (SNPs), G861C, G261T, and C129T, in the 5-HT1B gene. The goal was to investigate their association with the disease, to measure the strength of linkage disequilibrium (LD) between the SNPs, and to compare haplotype frequencies between alcoholic groups and normal controls. Data was also analyzed on the basis of Type I (n = 47) and Type II (n = 85) alcoholism. There was no significant difference in the allele frequencies or the genotype distribution between any alcoholic groups, alcoholic subgroups, and controls for any polymorphism. G861C and C129T polymorphisms were in complete LD. The pattern of distribution of haplotypes was similar in patients and controls. It is concluded that these SNPs are not playing any direct role in the development of susceptibility to alcoholism in our patient sample. PMID- 12898581 TI - Multipoint and single point non-parametric linkage analysis with imperfect data. AB - We used simulation to explore the impact of common data imperfections (i.e., missing parents, genotyping error, map error, and missing genotypes) upon the performance of multipoint and single point linkage analysis in the analyses of linkage data from pairs of siblings affected with an idealized complex trait. The performance of single point and multipoint linkage was similar under an unrealistic best case scenario; however, when four data imperfections were combined, the performance of single point linkage analysis appeared to be superior to multipoint. The absence of parental genotypes in the presence of 1% genotype error led to marked degradation of linkage signal, particularly for multipoint analyses. PMID- 12898582 TI - Bayesian analysis of a previously published genome screen for panic disorder reveals new and compelling evidence for linkage to chromosome 7. AB - This article presents a Bayesian re-analysis of a linkage study of panic disorder Crowe et al. [2001: Am J Med Genet (Neuropsychiatr Genet) 105:105-109]. In the initial analysis Crowe et al. failed to find compelling evidence for linkage based on either LOD scores or NPL scores anywhere in the genome. The maximum LOD score was 2.23 on chromosome 7 at marker D7S2846 (57.79 cM according to Marshfield). Over the past several years we have been developing a Bayesian alternative approach to linkage analysis, based on direct measurement of the posterior probability of linkage (PPL), and have shown elsewhere that this approach has several advantages over the available alternatives for mapping complex-disease genes Vieland [1998: Am J Med Genet 63:947-954]; Wang et al. [1999: Genet Epidemiol 17(Suppl 1):S749-S754]; Wang et al. [2000: Ann Hum Genet 64:533-553]; and Vieland et al. [2001: Hum Hered 51:199-208]. One limitation of this approach in previous applications has been that it required the investigator to specify a fixed genetic model for the trait. We employ a new implementation of the PPL that treats the unknown trait model as a vector of nuisance parameters, which is integrated out of the PPL equation. When we apply this new model integrated version of the PPL to the data of Crowe et al. [2001: Am J Med Genet (Neuropsychiatr Genet) 105:105-109] we obtain much clearer evidence than previously reported for a locus on chromosome 7, with an 80% probability of linkage to marker D7S521. A second location is also identified on chromosome 16 near marker D16S749 (PPL = 24%). The results for the remainder of the genome are consistently low. The two loci identified here are also supported by independent evidence from other studies. PMID- 12898584 TI - Children at familial risk for Tourette's disorder: Child and parent diagnoses. AB - Tourette's disorder (TD) is a neuropsychiatric disorder that onsets in childhood and appears to result from an interaction of genetic and environmental factors. The goal of the current prospective longitudinal study is to examine the onset of TD in offspring of parents with TD. At-risk children and control children were enrolled between the ages of 3 and 6 years and prospectively studied with yearly structured assessments over intervals of 2-5 years. Of 34 at-risk children free of tics at baseline, 10 (29%) subsequently developed a tic disorder, including 3 with TD. None of the 13 control children developed a tic disorder (P = 0.045). OCD or subclinical OCD emerged in 11 cases but no controls (P = 0.021). Finally, ADHD occurred in 14 at-risk children but no controls (P = 0.005). When at-risk families were dichotomized into bilineal or unilineal categories, rates of TD in children with two affected parents were three times the rate observed for children with one affected parent. These findings support the hypothesis that at least some cases of TD result from a complex genetic etiology. TD, OCD, and ADHD are significantly increased in children with TD parents. Furthermore, the risk is even greater when both parents are affected. Finally, our diagnosis of tic disorders in 21% of subjects initially reported by parents to be unaffected affirms the value of structured clinical assessments. PMID- 12898583 TI - Analysis of transmission of novel polymorphisms in the somatostatin receptor 5 (SSTR5) gene in patients with autism. AB - Infantile autism is a pervasive developmental disorder with a strong genetic component. The mode of inheritance appears to be complex and no specific susceptibility genes have yet been identified. Chromosome 16p13.3 may contain a susceptibility gene based on findings from genome scans and reports of chromosome abnormalities in individuals with autism. The somatostatin receptor 5 (SSTR5) gene is located on chromosome 16p13.3 and is thus a positional candidate gene for autism. SSTR5 may also be a functional candidate gene for autism because somatostatin inhibits growth hormone secretion, and increased growth hormone response has been reported in some individuals with autism. Moreover, the somatostatinergic system interacts with the dopaminergic system, which has been hypothesized to be involved in the etiology of autism; in particular, somatostatin secretion is regulated by dopamine, and the dopamine D2 receptor and the SSTR5 receptor interact to form a receptor complex with enhanced functional activity. In the present study, we tested whether the alleles of twelve new single nucleotide polymorphisms (SNPs) in the SSTR5 gene were preferentially transmitted, using the transmission disequilibrium test (TDT) in a sample of 79 trios with autism (18 from Denmark and 61 from France). Furthermore, we combined four missense SNPs into haplotypes and searched for preferential transmission using the program TRANSMIT. No significant preferential transmission of the alleles and haplotypes of the twelve SNPs was found. Our results do not suggest the SSTR5 gene as a susceptibility gene for autism. PMID- 12898585 TI - Extended investigation of tau and mutation screening of other candidate genes on chromosome 17q21 in a Swedish FTDP-17 family. AB - Frontotemporal dementia and Parkinsonism linked to chromosome 17 (FTDP-17) is an autosomal dominant condition clinically characterized by behavioral, cognitive and motor disturbances. It was recently discovered that the majority of the FTDP 17 families carry missense or 5' splice mutations in the exons coding for the microtubule-binding domains of the tau protein. However, in at least five FTDP-17 families, no such mutations could be identified. In the present study, we aimed at further investigate abnormalities in the tau gene in a Swedish FTDP-17 family, where no mutations in the tau gene previously have been identified. Initially, we searched for larger deletions by Southern blot hybridization. Furthermore, possible abnormal splicing events was investigated by RT-PCR from brain tissue of affected individuals. In addition, we investigated the presence of mutations in other genes in the FTDP-17 candidate region on chromosome 17q21; Gamma-tubulin, Glial Fibrillary Acid Protein (GFAP), Human dual specificity phosphatase tyrosine/serine (VHR), Rap-interacting protein 8 (RPIP8), P35, and the recently identified FTDCG1. In conclusion, no pathological changes in the tau gene were observed, neither was any mutations segregating with the disease detected in the investigated candidate genes. Further investigation of extended intron sequences or promoter regions of the tau gene and additional candidate genes on chromosome 17q21, therefore seems to be necessary in order to identify the additional causes of FTDP-17. PMID- 12898586 TI - Clinical features of boys with fragile X premutations and intermediate alleles. AB - Fragile X syndrome has a characteristic behavioural and physical phenotype. Clinical experience and case reports suggest that boys with premutations and intermediate alleles may have similar, but possibly milder, clinical features than those with the full mutation. We conducted detailed physical, psychiatric, psychological and speech and language evaluations on a clinical series of 10 boys, with either premutation or intermediate CGG triplet expansions. Wherever possible we measured the levels of FMR1 protein in participants' hair roots. Many participants demonstrated striking resemblance in their clinical picture, behavioural and physical, to individuals with the fragile X syndrome full mutation. However, protein expression was normal in all participants where it was assessed, despite large variation in CGG triplet repeats. We propose that the findings are unlikely to be attributable to ascertainment bias alone. Replication on larger independent samples is required to confirm our impression that fragile X premutations and intermediate alleles may be associated with important developmental disabilities and physical features. PMID- 12898587 TI - Association of the C766T polymorphism of the low-density lipoprotein receptor related protein gene with Alzheimer's disease. AB - The low-density lipoprotein receptor-related protein (LRP) is one of the most important cholesterol receptors in the brain. Gene variation of its ligand, apolipoprotein E, is a major genetic risk-factor for Alzheimer's disease (AD). The C-allele of the silent C766T polymorphism in exon 3 of the LRP gene might be associated with AD, however, results are conflicting and thus discussed controversially. Consequently, we compared the prevalence of this polymorphism in a homogenous cohort of patients with AD and control subjects. We found that carriers of a C-allele were at lower risk of AD; in agreement with this observation, AD patients who were carriers of a C-allele presented with a later age at onset of the disease than carriers of the TT genotype. These data suggest that LRP polymorphism influences the risk as well as the age at onset of AD. Our results contrast with other studies which described the C-allele to be a risk factor for AD, but are in line with a recent publication on the effect of LRP polymorphism on longevity and on the risk for coronary artery disease. Further research on LRP polymorphisms is needed to evaluate their effects on the risk of AD, on coronary artery disease and on longevity. PMID- 12898588 TI - Tissue-based stem cells: ABC transporter proteins take centre stage. AB - In adult tissues, with the notable exception of the haematopoietic system, the identity of somatic stem cells has often been inferred based on no more evidence than their location. Functional data (clonogenicity) to confirm such an appellation has been absent due to the inability to find appropriate cellular markers with which to isolate live stem cells. Flow cytometry has isolated haematopoietic stem cells based on their ability to exclude Hoechst dye. Named the side population (SP), this phenotype may be a universal marker of stem cells in adult tissues. PMID- 12898589 TI - Lymphatics and vascular channels; the differences behind similarity. AB - Although sharing many features and functions, there is a mounting body of evidence to suggest that there are subtle differences between lymphatics and vascular channels. New data suggest that Ets-1 and Ets-1-target MMP genes are differentially expressed in lymphatic and vascular channels. Since it is generally accepted that the Ets family of transcription factors play a significant role in the regulation of the expression of angiogenic factors, the finding that Ets-1 is apparently not involved in lymphangiogenesis may highlight one aspect where angiogenesis and lymphangiogenesis are differently regulated. PMID- 12898590 TI - High expression of MDR1, MRP1, and MRP3 in the hepatic progenitor cell compartment and hepatocytes in severe human liver disease. AB - An increase in bile ductular structures is observed in diverse human liver diseases. These structures harbour the progenitor cell compartment of the liver. Since ATP-binding cassette (ABC) transporters may have a cytoprotective role in liver disease, an immunohistochemical study was performed on human liver specimens from patients with primary biliary cirrhosis (PBC), chronic hepatitis C virus (HCV) infection, submassive cell necrosis, and normal liver. The expression of MDR1, MDR3, BSEP, MRP1, MRP2, and MRP3 was determined using specific antibodies. Dilution series were constructed to determine the critical staining level in order to estimate the factor of up-regulation. In normal liver, hepatocytes showed canalicular staining for MDR3, BSEP, and MRP2. MDR1 stained the canalicular membrane of hepatocytes as well as that of cholangiocytes. MRP3 showed low immunoreactivity of bile duct epithelial cells and centrilobular hepatocytes only. Normal liver showed no immunoreactivity for MRP1. In diseased liver, the expression of MDR3, BSEP, and MRP2 was relatively stable. In PBC, HCV, and submassive necrosis, the expression levels of MDR1, MRP1, and MRP3 were increased. The strongest immunoreactivity was seen after submassive necrosis, where remaining islands of hepatocytes showed strong canalicular staining for MDR1 and MRP3. Regenerating bile ductules at the interface of portal tracts and necrotic areas stained intensely for MDR1, MRP1, and MRP3. In conclusion, MDR1, MRP1, and MRP3 are up-regulated in hepatocytes in severe human liver disease. Strong MDR1, MRP1, and MRP3 reactivity is seen in regenerating human bile ductules. PMID- 12898591 TI - Ets 1 is expressed in capillary blood vessels but not in lymphatics. AB - Little is known about the mechanisms that regulate lymphangiogenesis and, in particular, about regulation at the transcriptional level. To determine whether parallels exist in the mechanisms of angiogenesis and lymphangiogenesis, the expression of the Ets 1 transcription factor (which has previously been shown to be involved in angiogenesis) was examined in two transgenic mouse lines, namely RipVEGF-C mice (which develop lymphatic capillaries ectopically around islets of Langerhans) and Rip1Tag2 mice (which develop insulinomas that are not lymphangiogenic). Crossing the two lines results in double transgenic mice that develop lymphatics around insulinomas, which in turn promotes metastasis to regional lymph nodes. By immunohistochemistry, it was found that, in contrast to blood vessels, lymphatic vessels in wild-type and transgenic mice did not express Ets 1. Immunohistochemical staining for matrix metalloproteinase (MMP)-2 and MMP 9 as well as membrane type 1-MMP (MT1-MMP/MMP-14), all of which are encoded by known or potential Ets 1 target genes, showed the same cellular distribution as Ets 1. These findings suggest that the transcriptional regulatory mechanisms of lymphangiogenesis differ from those involved in angiogenesis. Furthermore, if proteases are involved in lymphangiogenesis, these observations suggest that they are different from those considered to be important for blood vessel formation. PMID- 12898592 TI - Association of ets-related transcriptional factor E1AF expression with tumour progression and overexpression of MMP-1 and matrilysin in human colorectal cancer. AB - Expression of E1AF/PEA3 (ETV4), an ets family transcription factor, has been implicated in the invasive potential of several cancer cell lines through induction of matrix metalloproteinase (MMP) expression. The aim of this study was to examine E1AF mRNA expression and to determine whether it is correlated with progression of, and/or MMP expression in, human colorectal cancer. Using the semi quantitative reverse transcriptase-polymerase chain reaction (RT-PCR), 100 colorectal cancer tissues were analysed for E1AF mRNA expression. Expression of ER81 (ETV1) and ERM (ETV5), the other two members of the PEA3 subfamily, and Ets 1 and Ets-2 was also analysed. The results were correlated with clinicopathological characteristics and MMP expression. Immunohistochemical analysis and an in vitro invasion assay were also performed. E1AF mRNA expression was detected in 62% of the 100 colorectal cancer tissues, but was undetectable or only faintly detected in adjacent non-tumour tissues. E1AF mRNA was detected in all of the ten liver metastases from colorectal cancers. E1AF expression correlated significantly with depth of invasion, lymphatic and venous invasion, lymph node and distant metastasis, advance in pathological tumour-node-metastasis stage, and recurrence. Patients with E1AF-positive tumours had significantly shorter overall and disease-free survival periods than did those with E1AF negative tumours (p < 0.0001 and p < 0.0001, respectively). E1AF expression retained its significant predictive value for overall and disease-free survival in multivariate analysis that included conventional clinicopathological factors (p = 0.0066 and p = 0.0109, respectively). Among the MMPs analysed, expression of MMP-1 and matrilysin correlated significantly with E1AF expression. In contrast, expression of ER81 and ERM did not correlate with clinicopathological characteristics or the expression of these MMPs. Immunohistochemical expression of E1AF was predominantly observed at the invasive front, where the expression of MMP-1 and matrilysin and nuclear beta-catenin expression were often co-localized. Antisense E1AF-transfected HT-29 colon cancer cells expressed reduced levels of MMP-1 and matrilysin and were less invasive in vitro than neo-transfected HT-29 cells. The results of this study suggest that E1AF, the expression of which is closely correlated with the expression of MMP-1 and matrilysin, plays a key role in the progression of colorectal cancer. PMID- 12898593 TI - Quantitative analysis of colorectal tissue microarrays by immunofluorescence and in situ hybridization. AB - The accuracy and reliability of in situ studies may be compromised by qualitative interpretations. Quantitation imposes a greater degree of objectivity, is more reproducible, and facilitates the clarity of definitions. The aim of this study was to validate the utility of laser imaging systems for the in situ quantitative analysis of gene expression in tissue microarrays. Immunofluorescence was employed to quantify the expression of the tumour suppressor p53, a marker of proliferation (Ki67), an endothelial cell marker (CD31), and the mismatch repair proteins human Mut L homologue 1 and human Mut S homologue 2 in an arrayed series of colorectal tissues (n = 110). Quantitative data on this panel of antigens were compared objectively with qualitative scoring of immunohistochemical chromogen deposition. In addition, the expression of vascular endothelial growth factor (VEGF)-A, placental growth factor, hepatocyte growth factor, and c-Met mRNA was quantified by phosphor image analysis of in situ hybridization reactions. The quantified data on p53, Ki67, and CD31 expression were significantly associated with the pathologist's score (p < or = 0.001). While hepatocyte growth factor and placental growth factor were not up-regulated, c-Met expression was increased up to 2.5-fold and the median VEGF-A expression was elevated 4-fold (p = 0.003) in this series of colorectal tumours. Laser imaging systems are therefore feasible for high-throughput, quantitative profiling of tissue microarrays. PMID- 12898594 TI - Nuclear localization of the metastasis-related protein S100A4 correlates with tumour stage in colorectal cancer. AB - A large number of experimental studies have linked the S100A4 gene product to the metastatic phenotype of cancer cells and clinical evidence indicates a correlation between S100A4 expression and poor prognosis in several cancer types. The aim of the present study was to analyse the expression of the S100A4 protein in colorectal cancer. Paraffin-embedded samples from 277 colorectal cancer patients were immunostained with anti-S100A4 antibody. Cytoplasmic staining was observed in 178 of 277 samples (64%), whereas, unexpectedly, nuclear expression of S100A4 was found in 88 of 277 of the samples (32%). This novel finding was confirmed by western blot analysis of nuclear fractions isolated from frozen tumour tissue. Statistical analysis revealed a significant correlation between nuclear expression of S100A4 and tumour stage at diagnosis, while there was no such correlation between cytoplasmic staining and tumour stage. The nuclear localization of S100A4 in colorectal cancer and its relationship to tumour stage suggest that this protein may be involved in gene regulatory pathways of relevance to the metastatic phenotype of cancer cells. PMID- 12898595 TI - Immunoreactivity for cyclin D3 is frequently detectable in high-grade primary gastric lymphomas in the absence of the t(6;14)(p21.1;q32.3) chromosomal translocation. AB - Cyclin D3 plays a pivotal role in controlling the physiological progression from the G1 to the S phase of the cell cycle. Recent data suggest that cyclin D3 may be deregulated in extranodal non-Hodgkin's lymphomas (NHLs) as a consequence of the t(6;14)(p21.1;q32.3) translocation. The present study investigated for the first time by dual-colour fluorescence in situ hybridization (FISH) on interphase nuclei and immunohistochemistry the prevalence of the t(6;14) translocation and cyclin D3 immunoreactivity (IR) in a series of 29 stage I-IIE primary gastric NHLs (PGLs). No case showed the t(6;14) translocation. However, in five (17.2%) cases (two extranodal marginal zone lymphomas of MALT type, LGM; one diffuse large-cell lymphoma with a MALT component, DLCLM; and two diffuse large-cell lymphomas without a MALT component, DLCL), three to four cyclin D3 signals were detected by FISH. Co-hybridization with probes specific for the centromeric region and long arm of chromosome 6 indicated trisomy in one case (DLCL), whereas in the remaining four cases the pattern was highly suggestive of the presence of an isochromosome 6p. One (12.5%) case of LGM, six (75%) cases of DLCLM, and seven (53.8%) cases of DLCL (p = 0.0378) were immunoreactive for cyclin D3. Cyclin D3 IR was detected in two (40%) of the five cases with extra cyclin D3 signals and in 12 of the remaining 24 cases (50%, p = 1.000). These results suggest that the t(6;14) may represent a rare event in the pathogenesis of PGL and that cyclin D3 deregulation is most likely the result of epigenetic mechanisms. PMID- 12898596 TI - Raman spectroscopy, a potential tool for the objective identification and classification of neoplasia in Barrett's oesophagus. AB - Histopathology remains the gold standard technique for the diagnosis of intraepithelial neoplasia (dysplasia) in Barrett's oesophagus, but it is highly subjective and relies on blind biopsy targeting. The aim of this study was to evaluate Raman spectroscopy, a rapid, non-invasive, molecular, specific analytical technique, for the objective identification and classification of Barrett's neoplasia in vitro. A secondary objective was to demonstrate the need for a rigorous gold standard in the development of new diagnostic techniques. Forty-four patients with a mean age of 69 years (range 34-89 years) undergoing surveillance for Barrett's oesophagus were included in the study. Three consultant pathologists independently assessed snap-frozen oesophageal biopsy specimens. Raman spectra were measured on 87 histopathologically homogeneous samples. Spectral classification models were developed using multivariate analysis for the prediction of pathology. Histopathology and Raman classification results were compared. Raman spectral prediction with a consensus pathology classification model gave sensitivities between 73% and 100% and specificities of 90-100%. A high level of agreement (kappa = 0.89) was demonstrated between the three-subset biopsy targeting model and consensus pathology opinion. This compares favourably with the agreement measured between an independent pathologist and the consensus pathology opinion for the same spectra (kappa = 0.76). Raman spectroscopy appears to provide a highly sensitive and specific technique for the identification and classification of neoplasia in Barrett's oesophagus. PMID- 12898597 TI - Expression of p53, pRB, and p16 in lung tumours: a validation study on tissue microarrays. AB - Tissue microarrays have been created from 326 lung tumours, including 173 squamous cell carcinomas (SCCs) and 132 adenocarcinomas (ADs). In order to evaluate the usefulness of this microarray series, the expression of p53, p16, and Rb proteins was compared by immunohistochemistry on both the tissue microarrays and the corresponding whole sections for all 326 tumours. The presence of replicate punches improved both the yield and the concordance of data relative to the whole section results, so that the consensus score from the replicates agreed with the whole section result in more than 90% of informative tumours. The large number of tumours in this series also allowed significant differences in protein expression patterns to be detected between SCC and AD, the major subtypes of non-small cell lung carcinoma (NSCLC). SCC had higher levels of p53 staining (67% vs 52% in AD) and substantially increased p16 loss (SCC 75%, AD 53%) combined with greater retention of pRB expression (SCC 86% vs 67% in AD). The strong inverse correlation between p16 and pRB seen in SCC was essentially absent in AD. This study represents the largest single immunohistochemical survey of protein expression for p53, p16, and RB in NSCLCs. PMID- 12898598 TI - Survivin gene expression in early-stage non-small cell lung cancer. AB - Survivin is an inhibitor of apoptosis protein, overexpressed in most human malignancies and implicated in mitosis regulation and preservation of cell viability. In order to investigate the prevalence and clinical significance of survivin in early-stage non-small cell lung carcinoma (NSCLC), survivin mRNA levels and protein expression were evaluated, using quantitative real-time RT-PCR and immunohistochemistry, respectively, in a series of 83 patients with stage I (IA and IB) surgically resected NSCLC. Detectable survivin mRNA levels could be demonstrated in all non-neoplastic lung tissue samples and in the tumours analysed. Survivin mRNA levels were elevated in 80 carcinomas (96%) compared to normal lung (p = 0.008). Among all tumours, survivin transcripts were present at a higher level in squamous cell carcinomas (p = 0.0022). Cytoplasmic and nuclear immunoreactivity was found in 70% and 80% of tumours, respectively and both were present in 54%. Cytoplasmic immunoreactivity correlated with tumour stage (p = 0.019). Survivin expression levels did not correlate with patient survival. In one specimen, cytoplasmic and focal nuclear immunostaining was observed in dysplastic bronchial squamous metaplasia. These results document that survivin overexpression is almost always present in early-stage NSCLC, suggesting that this protein may play a role in lung tumourigenesis. This ubiquitous expression makes survivin an appealing new target for novel therapies in lung cancer. In addition, this study also documents that survivin overexpression could be exploited for diagnostic purposes and that quantitative real-time RT-PCR can be a useful tool for evaluating survivin activation in NSCLC. PMID- 12898599 TI - Beta-catenin abnormalities and associated insulin-like growth factor overexpression are important in phyllodes tumours and fibroadenomas of the breast. AB - The aim of this study was to assess the expression of IGF-I and IGF-II in phyllodes tumours and fibroadenomas and to see if there is any correlation between nuclear beta-catenin expression and IGF-I and IGF-II expression in these tumours. In a previous study, it has been shown that Wnt signalling is important in the pathogenesis of phyllodes tumours of the breast. Epithelial Wnt5a overexpression and stromal Wnt2 overexpression were associated with abnormal, nuclear localization of beta-catenin in the stromal cells of these tumours. However, not all tumours with beta-catenin accumulation showed Wnt overexpression. One other possible cause of beta-catenin accumulation is overexpression of insulin-like growth factors (IGFs), as both IGF-I and IGF-II have been shown to stabilize beta-catenin. In this study, 30 fibroadenomas of the breast were assessed for beta-catenin expression using immunohistochemistry and the results were compared with previous data from 119 phyllodes tumours. In situ hybridization was used to assess IGF-I and IGF-II expression in 23 phyllodes tumours and 16 fibroadenomas. Nineteen phyllodes tumours (83%) showed widespread overexpression of IGF-II and 5/23 (22%) showed widespread overexpression of IGF I. IGF-I expression correlated with nuclear beta-catenin staining in phyllodes tumours. Malignant phyllodes tumours showed no or little IGF-I expression. There was a degree of nuclear beta-catenin expression in the stroma (weak in 33%, moderate in 27%, and strong in 40%) in all fibroadenomas and nuclear beta-catenin staining correlated with IGF-I overexpression. Extensive IGF-II overexpression was also found in the majority of fibroadenomas (12/16). These results support the hypothesis that IGF-I and IGF-II overexpression may be important in the pathogenesis of fibroepithelial neoplasms of the breast and that IGF-I overexpression is likely to be contributing to the nuclear beta-catenin localization observed in the tumours. PMID- 12898600 TI - Loss of CD59 expression in breast tumours correlates with poor survival. AB - CD59 (protectin), a phosphatidylinositol-anchored glycoprotein, is a member of the cell membrane-bound complement regulatory proteins that inhibits the formation of the terminal membrane attack complex (MAC) of complement. In this study, the expression of CD59 was evaluated in 520 breast carcinomas from patients with a mean follow-up of 87 months. This expression was correlated with clinicopathological features and patient survival. Marked variation in the intensity of CD59 expression, which correlated with histological grade and Nottingham prognostic index (NPI), was found, with higher expression of CD59 found more often in well and moderately differentiated tumours and those of good prognosis (NPI < or = 3.4). In contrast, high grade and poor prognosis (NPI > 5.4) carcinomas significantly demonstrated lack of CD59 expression (p < 0.001). Moreover, it was found that the percentage of CD59-positive cells correlated significantly with patient survival, ie patients with a high percentage of positive cells (>50%) had a better overall survival (p = 0.006). A correlation was also found between the percentage of CD59-positive cells and tumour type and also the development of distant metastases. No association was found between either the intensity or the percentage of cells expressing CD59 and vascular invasion, lymph node stage, tumour size, patient age or menopausal status. In multivariate analysis, CD59 percentage positivity was of independent prognostic significance with grade and lymph node stage. These findings indicate that loss of CD59 may offer a selective advantage for breast cancers, resulting in more aggressive tumours and conferring a poor prognosis for patients. PMID- 12898601 TI - Loss of PDCD4 expression in human lung cancer correlates with tumour progression and prognosis. AB - The programmed cell death 4 gene (PDCD4), a newly identified transformation suppressor, was analysed in lung tumour cell lines and primary lung carcinomas. Reduced PDCD4 mRNA expression was observed in two immortalized lung cell lines and 18 cancer cell lines by northern blot analysis. In the survey of primary lung tumours, PDCD4 cDNA was poorly represented in 47 lung tumours compared with normal lung tissue by cDNA microarray analysis and this poor representation was significantly associated with high-grade (G3) adenocarcinomas (p = 0.012). Immunohistochemical analysis of 124 primary carcinomas comprising all subtypes demonstrated that PDCD4 protein expression was widely lost in tumour samples (83%) and was negatively related to poor prognosis (p = 0.013). The loss of PDCD4 expression correlated with higher grade and disease stage (p = 0.045 and 0.034, respectively), but not tumour size and nodal status. Similarly to the cDNA data, lack of PDCD4 expression was significantly linked to tumour grade in adenocarcinoma (n = 59, p = 0.048), while in squamous cell carcinoma (n = 58), no relationship between PDCD4 expression and clinicopathological parameters was established. These data suggest that the loss of PDCD4 expression is a prognostic factor in lung cancer and may correlate with tumour progression. PMID- 12898602 TI - Expression of nitric oxide synthase isoforms in human liver cirrhosis. AB - Several mediators of systemic vasodilatation in liver cirrhosis have been reported. Among these is nitric oxide (NO), which has been proposed as one of the main mediators. In this study, sera and liver biopsies were analysed from 15 patients with clinically and pathologically diagnosed liver cirrhosis. In addition, sera from seven and liver biopsies from three healthy controls were used. Serum levels of nitrite (the end product of NO) were measured using the Griess reaction and the expression of the inducible nitric oxide synthase (iNOS) and constitutive nitric oxide synthase (ecNOS) proteins was investigated using immunohistochemistry. This study shows that serum nitrite levels (94 +/- 9.8 micro mol/l) in cirrhotic patients were significantly (p < 0.05) increased in comparison with the controls (36.6 +/- 11.03 micro mol/l). iNOS was completely absent from the control group but was highly expressed in the livers from the cirrhotic group. iNOS was seen mainly in the inflammatory cells infiltrating the portal tracts, blood monocyte-like cells, hepatocytes, sinusoidal cells, and endothelial cells. However, expression of ecNOS was only seen in the vascular endothelial cells of both the control and the cirrhotic groups, but was much higher in the latter. It is therefore clear that NO is augmented in cirrhotic patients and it is mainly produced by induction of iNOS. Moreover, NO up regulation is dependent on the inflammatory stage of liver cirrhosis. ecNOS production could be a normal chronic adaptation mechanism of the endothelium to the chronically increased splanchnic blood flow secondary to portal hypertension. In the near future, the appropriate inhibition of NO activity by using NOS-active agents may provide a novel strategy for the treatment of patients with liver cirrhosis. PMID- 12898603 TI - Typical genomic imbalances in primary MALT lymphoma of the orbit. AB - Primary orbital non-Hodgkin lymphoma is a mucosa-associated lymphoid tissue (MALT)-type extranodal marginal zone lymphoma. Little information is available on its genome as conventional cytogenetics is limited by scarce biopsy material, while fluorescence in situ hybridization (FISH) explores only selected regions. Comparative genomic hybridization (CGH) performs full genomic analysis and is applicable to different sources of DNA, such as fresh and frozen cells, as well as paraffin-embedded tissues. In this study, CGH was used to analyse primary MALT lymphoma of the orbit. Aneuploidy was identified in six of the ten cases studied. Gains (19) were more frequent than losses (5). The most frequent duplications involved chromosome 3 (common region at 3q24-qter), as expected in marginal zone lymphoma, and chromosome 6 (common region at 6p21.1-21.3), which is typical of an orbital location. Other chromosome gains were found at 1p, 7, 8q, 9q, 12, 13, 17, 18, 19, 22, and X. Losses were located at 1q, 6q, 9q, 11q, and 13q. Two cases showed isolated duplications of chromosome 6p or 9q. Isolated imbalances were found only in tumours affecting the conjunctiva. Complex aneuploidies were observed in lymphoma of the retro-orbital tissue. In summary, CGH in orbital MALT lymphoma provided new insights into typical genomic imbalances and underlying pathogenetic mechanisms. PMID- 12898604 TI - Measurement of telomere length on tissue sections using quantitative fluorescence in situ hybridization (Q-FISH). AB - Loss of telomere repeat sequences occurs after each cell division and telomere shortening has been implicated in cellular senescence. The measurement of telomere length might therefore assess the lifespan of a cell. The aim of this study was to set up and validate a technique enabling the assessment of telomere length on tissue sections. Quantitative fluorescence in situ hybridization (Q FISH) with telomeric probes was performed on smears and sections from cell preparations or human tissues. The mean fluorescence intensity of telomere spots (FI/spot) was automatically quantified by image analysis. Telomeric restriction fragment (TRF) length was assessed by Southern blotting. There was a positive significant correlation between telomere length, as assessed by Q-FISH, and TRF length determined by Southern blotting in corresponding samples (p < 0.01, r = 0.6 for tissue and p < 0.01, r = 0.79 for cells). FI/spot was higher on smears than on sections, but pairwise comparison showed a significant correlation both for cells and for tissues (r = 0.77, p < 0.001 for cells and p < or = 0.01, r = 0.64 for tissue). Finally, since telomere length is expected to shorten with age, FI/spot was assessed in liver samples according to the age of patients: a negative correlation was demonstrated (r = 0.76, p < 0.01). Inter-assay variation was 7% for Q-FISH performed on tissue sections and 12% on touch preparations. This study shows that Q-FISH can be performed with confidence on fixed frozen tissue sections in order to assess telomere length. It is an easy, accurate, and reproducible in situ method for assessing telomeres in the context of cell type and tissue architecture. PMID- 12898605 TI - The wt1-heterozygous mouse; a model to study the development of glomerular sclerosis. AB - In the present study, it is shown that mice heterozygous for wt1 develop glomerular sclerosis and the nature and time course of events leading to the glomerular scarring are determined. Wt1-heterozygous (wt1het) mice and their wild type littermates were closely monitored from birth and plasma levels of urea, creatinine, and albumin were compared with histological data and clinical features. One of the first indications of nephropathy in the wt1het mouse was the development of proteinuria, accompanied by progressive elevation of the plasma levels of urea and creatinine. Subsequently, the mice developed albuminuria, which correlated with thickening of the glomerular basement membrane and fusion of the podocyte foot processes. Glomerulosclerosis was a relatively late event, accompanied by severe albuminuria and loss of WT1, nephrin, CD2AP, and alpha actinin-4. PMID- 12898607 TI - Sorting and convergence of primary olfactory axons are independent of the olfactory bulb. AB - Primary olfactory axons expressing the same odorant receptor gene sort out and converge to fixed sites in the olfactory bulb. We examined the guidance of axons expressing the P2 odorant receptor when they were challenged with different cellular environments in vivo. In the mutant extratoes mouse, the olfactory bulb is lacking and is replaced by a fibrocellular mass. In these animals, primary olfactory axons form glomerular-like loci despite the absence of normal postsynaptic targets. P2 axons are able to sort out from other axons in this fibrocellular mass and converge to form loci of like axons. The sites of these loci along mediolateral and ventrodorsal axes were highly variable. Similar convergence was observed for larger subpopulations of axons expressing the same cell surface carbohydrates. The sorting out and convergence of like axons also occurred during regeneration following bulbectomy. Olfactory axon behaviour in these models demonstrates that sorting and convergence of axons are independent of the target, which instead provides distinct topographic cues for guidance. PMID- 12898606 TI - Vocal circuitry in Xenopus laevis: telencephalon to laryngeal motor neurons. AB - Sexually differentiated calling patterns of Xenopus laevis are conveyed to the vocal organ by a dedicated neuromuscular system. Here, we define afferents to vocal motor neurons and determine whether the connectivity of the vocal pathway is sexually differentiated. The use of fluorescent dextran amines and the isolated brain preparation readily permitted identification of anterograde and retrograde connectivity patterns. The whole-mount preparation allowed us to observe projections in their entirety, including cells of origin of a projection (for retrograde projections), terminal fields (for anterograde connections), and fiber tracts. Major findings are the confirmation of a robust and reciprocal connection between cranial nucleus (n.) IX-X and the pretrigeminal nucleus of the dorsal tegmental area of the medulla (DTAM) as well as between DTAM and the ventral striatum (VS). Newly revealed is the extensive connectivity between the rostral subdivision of the dorsal nucleus raphe (rRpd) and candidate vocal nuclei. In contrast to previous results using peroxidase, we did not observe dramatic sex differences in connectivity, although some connections were less robust in female than in male brains. Some retrograde connections previously observed (e.g., anterior preoptic area to DTAM) were not confirmed. Plausible hypotheses are that a set of rhombencephalic neurons located in DTAM, the inferior reticular formation and n.IX-X are responsible for generating patterned vocal activity, that activity is modulated by neurons in rRpd, and that activity in VS (particularly that evoked by conspecific calls), together with effects of steroid hormones at many sites in the vocal circuit, contribute to the initiation of calling. PMID- 12898608 TI - Pretectotectal pathway: an ultrastructural quantitative analysis in cats. AB - Both the pretectum (PT) and the superior colliculus (SC) play an important role in directing eye movements and in sensorimotor coupling. A reciprocal connection between the PT and the SC has been described, which suggests a strong interplay between these two structures. We injected the cat SC with retrograde tracers and examined the labeled pretectotectal (PTT) cells at the light and electron microscopic level. PTT cells were distributed mostly in the nucleus of the optic tract and 93.1% contained gamma amino butyric acid (GABA). We also observed that PTT cells are located outside of pretectal regions distinguished by dense retinal terminals and clusters of cells that contain calbindin. This suggests that the GABAergic PTT cells are distinct from the GABAergic pretectogeniculate cells that have been previously described as being distributed within these regions. Finally, to determine the synaptic targets of PTT terminals, we injected the PT with anterograde tracers and examined terminals labeled in the SC at the ultrastructural level. The labeled PTT terminals were beaded fibers that were distributed mainly within the stratum griseum superficiale (SGS) of the SC. Using postembedding immunocytochemistry, 94.5% were found to be GABAergic. The PTT terminals were mostly small in size and primarily contacted GABA-negative dendrites (88.1%) and in some cases somata (4.7%). The remainder terminated on GABAergic dendrites (7.2%). Our results suggest that the PTT cells constitute a separate population of GABAergic efferent cells in the PT, which may function to inhibit the activity of non-GABAergic SC efferent cells in the SGS. PMID- 12898609 TI - Differential patterns of glial fibrillary acidic protein-immunolabeling in the brain of adult lizards. AB - The present study describes by means of immunohistochemistry the comparative distribution of glial fibrillary acidic protein (GFAP)-positive cells in the forebrain and midbrain of three species of lizards: Eumeces algeriensis, Scincoidae; Agama impalearis, Agamidae; Tarentola mauritanica, Gekkonidae. In the species studied, the different types and proportions of glial cells expressing GFAP showed considerable variation. These cells include radial glia, oval cells, tanycytes, ependymocytes, glia limitans, and astrocytes. In Eumeces, astrocytes are particularly abundant and their processes form numerous perivascular end feet; in addition well-differentiated ependymal cells and glia limitans express GFAP. These mature glial features are concordant with the relatively advanced phylogenetic level of Eumeces. In Tarentola, relatively few GFAP-expressing glial cells are observed, consisting mainly of radial glia and tanycytes. These features indicate a relatively immature state of the glial cell populations in this species. In Agama, GFAP-immunostained cells are confined to the periventricular and subpial brain areas; the ventricular lining contains numerous GFAP-immunopositive tanycytes and well-differentiated glia limitans. This pattern indicates that the glial cell profile in Agama exhibits characteristics intermediate between Eumeces and Tarentola, a feature which is discordant with the relatively primitive phylogenetic level of Agamidae compared to Gekkonidae. Together, the results of the present study provide novel data on the characterization of GFAP-expressing cell populations in different species of lizards. We suggest that the different glial patterns observed in the lizard brain correlates with developmental and functional aspects. PMID- 12898610 TI - Interference with anoikis-induced cell death of dopamine neurons: implications for augmenting embryonic graft survival in a rat model of Parkinson's disease. AB - One promising therapy for the treatment of Parkinson's disease is transplantation of embryonic ventral mesencephalic tissue. Unfortunately, up to 95% of grafted cells die, many via apoptosis. In this study we attempted to prevent anoikis induced cell death, which is triggered during the preparation of cells for grafting, and examine the impact on graft viability and function. We utilized the extracellular matrix molecule tenascin-C (tenascin) and an antibody (Ab) to the cell adhesion molecule L1 to specifically mimic survival signals induced by cell matrix and cell-cell interactions. In vitro, both tenascin- and L1 Ab-treated cultures doubled the number of tyrosine hydroxylase immunoreactive (THir) neurons compared to control. Additionally, cell survival assays determined that tenascin and L1 Ab-treated cell suspensions yielded more metabolically active and fewer dead cells than control suspensions. In contrast to the culture results, tenascin and L1 Ab-treated mesencephalic grafts did not yield an increase in the number of THir neurons using our standard grafting paradigm (3 microl of 100,000 cells/microl). However, under low-density conditions (3 microl of 3,000 cells/microl), tenascin augmented grafted THir neuron survival. These findings are consistent with the view that cell density can dramatically influence the degree of stress placed on THir neurons and consequently affect the success of survival strategies in vivo. In conclusion, pretreatment with tenascin may prove beneficial to prevent anoikis in dilute cell suspension grafts, while long-term in vivo delivery methods need to be explored to determine if L1 can prevent anoikis in grafts of mesencephalic dopamine neurons after transplantation. PMID- 12898611 TI - Neuronal expression of tachykinin-related peptides and gene transcript during postembryonic development of Drosophila. AB - The gene Dtk, encoding the prohormone of tachykinin-related peptides (TRPs), has been identified from Drosophila. This gene encodes five putative tachykinin related peptides (DTK-1 to 5) that share the C-terminal sequence FXGXRamide (where X represents variable residues) as well as an extended peptide (DTK-6) with the C-terminus FVAVRamide). By mass spectrometry (MALDI-TOF-MS), we identified ion signals with masses identical to those of DTK-1 to 5 in specific brain regions. We have analyzed the distribution of the Dtk transcript and peptides, by in situ hybridization and immunocytochemistry during postembryonic development of the central nervous system (CNS) of Drosophila. Antiserum against a cockroach TRP that cross-reacts with the DTKs was used for immunocytochemistry. Expression of transcript and peptides was detected from first to third instar larvae, through metamorphosis to adult flies. Throughout postembryonic development, we were able to follow the strong expression of TRPs in a pair of large descending neurons with cell bodies in the brain. The number of TRP expressing neuronal cell bodies in the brain and ventral nerve cord increases during larval development. In the early pupa (stage P8), the number of TRP expressing cell bodies is lower than in the third instar larvae. The number drastically increases during later pupal development, and in the adult fly about 200 TRP-expressing neurons can be seen in the CNS. The continuous expression of TRPs in neurons throughout postembryonic development suggests specific functional roles in both larval and imaginal flies and possibly also in some neurons during pupal development. PMID- 12898612 TI - Translocation of presynaptic delta opioid receptors in the ventrolateral periaqueductal gray after swim stress. AB - Immunolabeling for the delta opioid receptor (DOR) is localized primarily to axon terminals in the ventrolateral periaqueductal gray (vlPAG). However, rather than on the plasma membrane, DOR immunoreactivity is usually located within the cytoplasmic compartment, often associated with dense-core vesicles. In this study, the hypothesis that a behavioral stimulus, a cold water swim stress (3 minutes at 4 degrees C; CWSS), could initiate the translocation of the DOR was tested. The subcellular distribution of DOR was examined using a preembedding immunogold-labeling method and ultrastructural analysis in control rats and in rats that had a CWSS. In both cases, dense-core vesicles associated with DOR labeling were often within 100 nm of the plasma membrane. When the dense-core vesicles were near the plasma membrane, sometimes electron-dense "tethers" appeared between the vesicle and the plasma membrane. However, in rats exposed to CWSS, there was a decrease in immunolabeling associated with dense-core vesicles that were near the plasma membrane and a significant increase in DOR immunoreactivity associated with the plasma membrane. In addition, there was a significant increase in the fraction of DOR immunoreactivity associated with large clear-core vesicles; possibly early endosomes. Moreover, after a CWSS, dense-core vesicles containing DOR immunoreactivity could be visualized fusing with the plasma membrane of synaptic boutons. These data suggest the involvement of DOR in the vlPAG in the behavioral response to CWSS. Furthermore, the results support the hypothesis that the cell surface distribution of presynaptic receptors can be regulated in an activity-dependent manner by virtue of transport via dense-core vesicles. PMID- 12898613 TI - Characterization of lacZ-expressing cells in the gut of embryonic and adult DbetaH-nlacZ mice. AB - In mice that express lacZ under the control of a human dopamine beta-hydroxylase gene promoter (DbetaH-nlacZ mice), the nuclei of enteric neurons express the transgene, as shown by the presence of beta-galactosidase (beta-gal) staining (Mercer et al. [1991] Neuron 7:703-716). The transgene is also expressed by neural crest-derived cells in the developing gut before their differentiation into neurons or glial cells (Kapur et al. [1992] Development 116:167-175). However, the cell types expressing the DbetaH-nlacZ transgene within the developing and adult gut have not been fully characterized. Whole-mount preparations of embryonic and adult gut were processed for histochemistry or immunohistochemistry to reveal beta-gal plus markers of undifferentiated neural crest cells (in embryos) or enteric neurons (in adults). In embryonic mice, over 90% of undifferentiated neural crest-derived cells (identified using antibodies to p75) were beta-gal(+). Importantly, crest-derived cells at the migratory wavefront were all beta-gal(+). In adult mice, only a subpopulation of enteric neurons was beta-gal(+), while glial cells showed no beta-gal staining. Considerable variation was observed between the small intestine and colon in the proportion of myenteric neurons that showed beta-gal staining. We examined whether known classes of enteric neurons varied in their expression of DbetaH nlacZ. In the myenteric plexus of the jejunum and colon, large calretinin(+) neurons did not express lacZ, suggesting that the incomplete penetrance of the DbetaH-nlacZ transgene observed in adult mice is not random. We conclude that the DbetaH-nlacZ transgene provides a reliable marker for examining the colonization of the developing gut by neural crest cells. However, in adult mice, there is variation between mice, between gut regions, and between different classes of enteric neurons in the expression of the transgene. PMID- 12898614 TI - Overlapping distributions of orexin/hypocretin- and dopamine-beta-hydroxylase immunoreactive fibers in rat brain regions mediating arousal, motivation, and stress. AB - A double-label immunohistochemical study was carried out to investigate overlap between dopamine-beta-hydroxylase (DbetaH) -immunopositive projections and the projections of hypothalamic neurons containing the arousal- and feeding-related peptide, orexin/hypocretin (HCRT), in rat brain. Numerous intermingled HCRT immunopositive and DbetaH-immunopositive fibers were seen in a ventrally situated corridor extending from the hypothalamus to deep layers of the infralimbic cortex. Both fiber types avoided the nucleus accumbens core, caudate putamen, and the globus pallidus. In the diencephalon, overlap was observed in several hypothalamic areas, including the perifornical, dorsomedial, and paraventricular nuclei, as well as in the paraventricular thalamic nucleus. Intermingled HCRT containing and DbetaH-containing fibers extended from the hypothalamus into areas within the medial and central amygdala, terminating at the medial border of the lateral subdivision of the central nucleus of the amygdala. Dense overlap between the two fiber types was also observed in the periaqueductal gray, particularly in the vicinity of the dorsal raphe, as well as (to a lesser extent) in the ventral tegmental area, the retrorubral field, and the pedunculopontine tegmental nucleus. Hypocretin-containing cell bodies, located in the perifornical and lateral hypothalamus, were embedded within a dense plexus of DbetaH immunopositive fibers and boutons, with numerous cases of apparent contacts of DbetaH-containing boutons onto HCRT-immunopositive soma and dendrites. HCRT containing fibers were observed amid the noradrenergic cells of the locus coeruleus, and in the vicinity of the A1, A2, and A5 cell groups. Hence, the projections of these two arousal-related systems, originating in distinctly different parts of the brain, jointly target several forebrain regions and brainstem monoaminergic nuclei involved in regulating core motivational processes. PMID- 12898615 TI - Dendritic domains of nociceptive-responsive parabrachial neurons match terminal fields of lamina I neurons in the rat. AB - This study investigates, in the anesthetized rat, the dendritic extent of parabrachial (PB) neurons whose nociceptive response to noxious stimuli has been previously recorded with an extracellular micropipette. The PB neurons were then injected with biocytin through the recording micropipette, via a juxtacellular technique. The dendritic arborization of individual PB neurons was carefully compared with the projections of medullary (trigeminal) and spinal lamina I neurons. The latter projections were labeled in separate animals that received injections of Phaseolus vulgaris-leucoagglutinin restricted to the superficial layers of spinal or medullary dorsal horn. We report here that: 1) PB neurons excited chiefly by noxious stimulation of the face have their dendritic tree located primarily within the field of lamina I trigeminal projections, i.e., in the caudal portion of PB area, around the external medial and the caudal part of the external lateral subnuclei; and 2) PB neurons excited chiefly by noxious stimulation of the paw or the tail have their dendritic tree located primarily within the field of lamina I spinal projections, i.e., in PB mid-extent, around the borderline between the external lateral and both the lateral crescent and the superior lateral subnuclei. Our results suggest the presence of an extensive excitatory axodendritic link between lamina I projections and PB nociceptive neurons around the lateral crescent and the external medial subnuclei. These findings strengthen the possibility of involvement of a subgroup of PB neurons in nociceptive processes. PMID- 12898617 TI - Nature's gift to science (Nobel lecture). PMID- 12898618 TI - Caenorhabditis elegans: the cell lineage and beyond (Nobel lecture). PMID- 12898620 TI - Macrophomate synthase: the first structure of a natural Diels-Alderase. PMID- 12898619 TI - Worms, life, and death (Nobel lecture). PMID- 12898621 TI - A stem cell molecular signature: are there hallmark properties that are shared by all stem cells? PMID- 12898622 TI - Alteration of the substrate specificity of benzoylformate decarboxylase from Pseudomonas putida by directed evolution. AB - Alteration of the substrate specificity of thiamin diphosphate (ThDP)-dependent benzoylformate decarboxylase (BFD) by error-prone PCR is described. Two mutant enzymes, L476Q and M365L-L461S, were identified that accept ortho-substituted benzaldehyde derivatives as donor substrates, which leads to the formation of 2 hydroxy ketones. Both variants, L476Q and M365L-L461S, selectively catalyze the formation of enantiopure (S)-2-hydroxy-1-(2-methylphenyl)propan-1-one with excellent yields, a reaction which is only poorly catalyzed by the wild-type enzyme. Different ortho-substituted benzaldehyde derivatives, such as 2-chloro-, 2-methoxy-, or 2-bromobenzaldehyde are accepted as donor substrates by both BFD variants as well and conversion with acetaldehyde resulted in the corresponding (S)-2-hydroxy-1-phenylpropan-1-one derivatives. As deduced from modeling studies based on the 3D structure of wild-type BFD, reduction of the side chain size at position L461 probably results in an enlarged substrate binding site and facilitates the initial binding of ortho-substituted benzaldehyde derivatives to the cofactor ThDP. PMID- 12898623 TI - Structural investigation of the HIV-1 envelope glycoprotein gp160 cleavage site, 2: relevance of an N-terminal helix. AB - Proteolytic activation of the HIV-1 envelope glycoprotein gp160 is selectively performed by the proprotein convertase furin at the C-terminus of the sequence R508-E-K-R511 (site 1), in spite of the presence of another consensus sequence, Lys500-Ala-Lys-Arg503 (site 2). On the basis of the solution structural analysis of the synthetic peptide p498, spanning the gp160 sequence Pro498-Gly516, we previously suggested a possible role of an N-terminal helix in regulating the exposure and accessibility of the gp160 physiological cleavage site, enclosed in a loop. Here we report on the activity and conformation of the 23-residue peptide h-REKR, designed to exhibit a large N-terminal helix, followed by the gp160 native sequence, Arg508-Gly516. h-REKR is digested by furin with high efficiency, comparable to the full native p498. Circular dichroism analyses, in mixtures from pure water to 98 % trifluoroethanol, outline a significant content of helical structure in the peptide conformation. The molecular model obtained from NMR data collected in trifluoroethanol/water, by means of DYANA and AMBER simulations, indeed has helical structure on a large N-terminal segment. Such a long helix does not seem to affect the loop conformation of the C-terminal site 1-containing sequence, which exhibits the same proton chemical shifts already observed for the full native p498. PMID- 12898624 TI - Protein stabilisation by compatible solutes: effect of mannosylglycerate on unfolding thermodynamics and activity of ribonuclease A. AB - Differential scanning calorimetry, optical spectroscopy, and activity measurements were used to investigate the effect of mannosylglycerate, a negatively charged osmolyte widely distributed among thermophilic and hyperthermophilic archaea and bacteria, on the thermal unfolding of ribonuclease A (RNase A). For comparison, assays in the presence of trehalose, a canonical solute in mesophiles, and potassium chloride were also carried out. A thermodynamic analysis was performed by using differential scanning calorimetry data. The changes in the heat capacity for unfolding were similar for the different solutes examined. Mannosylglycerate was an efficient thermostabiliser of RNase A and induced an increase of 6 degrees C mole(-1) in the melting temperature. Moreover, the performance of mannosylglycerate as a stabiliser depended on the net charge of the molecule, with the maximal effect being observed at pH values above 4.5. Analysis of the enthalpic and entropic contributions to unfolding, derived from calorimetric data, revealed that the stabilisation rendered by mannosylglycerate is primarily achieved through a decrease in the unfolding entropy. Also, the number of protons taken up by RNase A upon denaturation in the presence of mannosylglycerate was considerably higher than with other solutes, a result consistent with a more rigid structure of the native protein. Mannosylglycerate (potassium salt) inhibited the activity of RNase A, albeit to a smaller extent than KCl, and acted as an efficient suppressor of aggregation of the denatured protein, thereby having a remarkable beneficial effect on the inactivation of RNase A upon thermal denaturation. The results are discussed in view of the physiological role of this charged compatible solute. PMID- 12898625 TI - Comparative study of purine and pyrimidine nucleoside analogues acting on the thymidylate kinases of Mycobacterium tuberculosis and of humans. AB - Thymidine monophosphate kinase (TMPK) from Mycobacterium tuberculosis (TMPKmt) is an attractive target for the design of specific inhibitors. This fact is the result of its key role in the thymidine pathway and of unique structural features in the active site observed by X-ray crystallography, especially in comparison to its human counterpart (TMPKh). Different 5-modified thymidine derivatives, as well as purine and pyrimidine analogues or C-nucleosides were tested on TMPKmt and TMPKh, and the results were rationalized by docking studies. 5-Halogenated 2' deoxyuridines are the best inhibitors of TMPKmt found and present the highest selectivity indexes in favor of TMPKmt. PMID- 12898626 TI - Selection of D-amino-acid peptides that bind to Alzheimer's disease amyloid peptide abeta1-42 by mirror image phage display. AB - A mirror image phage display approach was used to identify novel and highly specific ligands for Alzheimer's disease amyloid peptide Abeta(1-42). A randomized 12-mer peptide library presented on M13 phages was screened for peptides with binding affinity for the mirror image of Abeta(1-42). After four rounds of selection and amplification the peptides were enriched with a dominating consensus sequence. The mirror image of the most representative peptide (D-pep) was shown to bind Abeta(1-42) with a dissociation constant in the submicromolar range. Furthermore, in brain tissue sections derived from patients that suffered from Alzheimer's disease, amyloid plaques and leptomeningeal vessels containing Abeta amyloid were stained specifically with a fluorescence labeled derivative of D-pep. Fibrillar deposits derived from other amyloidosis were not labeled by D-pep. Possible applications of this novel and highly specific Abeta ligand in diagnosis and therapy of Alzheimer's disease are discussed. PMID- 12898627 TI - Tamoxifen derivatives for delivery of the antitumoral (DACH)Pt group: selective synthesis by McMurry coupling, and biochemical behaviour. AB - The goal of our study was to potentiate the effects of the ((R,R)-trans-1,2 diaminocyclohexane)-platinum(II) fragment [(DACH)Pt], known for its cytotoxic properties, either with tamoxifen (Tam), the most widely used antiestrogen in the treatment of hormone-dependent breast cancers, or with its active metabolite hydroxytamoxifen (hydroxy-Tam). We coupled Tam or hydroxy-Tam derivatives bearing a malonato group at the para position of the beta aromatic ring with the (DACH)Pt fragment. The malonato-Tam and malonato-hydroxy-Tam compounds were prepared through McMurry coupling of the appropriate ketones. The presence of the malonate group resulted in a pronounced stereospecificity in the reaction, since malonato Tam was obtained only as the Z isomer, while malonato-hydroxy-Tam was obtained as an 80/20 E/Z mixture. Attribution of the isomeric structures was achieved by 2D NMR spectroscopy. The platinum complexes (DACH)Pt-malonato-Tam and (DACH)Pt malonato-hydroxy-Tam were then prepared by coupling the barium salts derived from the malonato-Tam and malonato-hydroxy-Tam with the nitrate derived from (DACH)PtCl(2). Study of the biochemical properties of these two platinum complexes showed that, while the hydroxy-Tam complex is satisfactorily recognized by the estrogen receptor (relative binding affinity, RBA=6.4 %), the Tam complex is less well recognized (RBA=0.5 %). The effects of these complexes on two hormone-dependent breast cancer cell lines (MCF7 and MVLN) were studied in vitro. Both complexes showed an antiproliferative effect on MCF7 cells, and an antiestrogenic effect on MVLN cells. The observed effects appear to be essentially antihormonal, since incorporation of the (DACH)Pt fragment into the tamoxifen skeleton did not cause an increase in the cytotoxicity of the complexes. PMID- 12898628 TI - Photoreversible inhibition of cholinesterases: catalytic serine-labeled caged butyrylcholinesterase. AB - The photoregulation of the catalytic activity of butyrylcholinesterase (BChE) was investigated by treating the enzyme with a newly developed carbamylating reagent, N-methyl-N-(2-nitrophenyl)carbamoyl chloride, which has proved to be an efficient photoremovable alcohol-protecting group. BChE was efficiently inhibited in a time and concentration-dependent manner, and the enzyme could be protected against inhibition by active-site-specific ligands (that is, tacrine). The inactivation kinetics showed a biphasic character, which can be analyzed as the result of the existence of two conformational states in solution. Pseudo-irreversible inactivation of BChE, which results from catalytic serine carbamylation, was suggested by recovery of the enzyme activity after dilution and was demonstrated by X-ray crystallography. Remarkably, the 3D structure of the carbamylated BChE conjugate showed a nonambiguous carbamylation of the catalytic serine residue as the only chemical modification on the protein. The photoreversibility of the enzyme inactivation was analyzed by irradiating the inactivated enzyme at 365 nm and was shown to reach completion in some conditions. The efficient and specific "caging" of BChE, together with the availability of carbamylated BChE crystals, will offer a unique possibility to study the catalytic properties of this enzyme by kinetic crystallography after cryophotolytic uncaging of the enzyme conjugate crystals. PMID- 12898629 TI - CloN6, a novel methyltransferase catalysing the methylation of the pyrrole-2 carboxyl moiety of clorobiocin. AB - The aminocoumarin antibiotic clorobiocin contains a 5-methylpyrrole-2-carboxylic acid unit. This pyrrole unit is derived from L-proline, and it would be expected that its 5-methyl group should be introduced by a methylation reaction. However, sequence analysis of the clorobiocin biosynthetic gene cluster did not reveal a gene with sequence similarity to the SAM-dependent methyltransferases that could be assigned to this reaction. This study, however, has provided evidence that the gene cloN6 is involved in this methylation reaction. Its gene product CloN6 shares conserved sequence motifs with the recently identified radical SAM protein superfamily, and it has been suggested that members of this family can catalyse methylcobalamin-dependent methylation reactions. cloN6 was inactivated in the clorobiocin producer Streptomyces roseochromogenes var. oscitans DS 12.976 by use of the PCR-targeting method. The cloN6(-) mutants accumulated, instead of clorobiocin, a derivative lacking the 5"'-methyl group of the pyrrole moiety (termed novclobiocin 109). A structural isomer carrying the pyrrole-2-carboxyl moiety at 2"-OH rather than at the 3"-OH of the deoxysugar (novclobiocin 110), and a derivative completely lacking the pyrrole unit (novclobiocin 104) were also identified. The structures of the metabolites were confirmed by NMR and MS analysis. Antibacterial activity tests against Bacillus subtilis showed that novclobiocin 109 and novclobiocin 110 have antibacterial activities about eight times less than that of clorobiocin, whereas novclobiocin 104 showed no activity under the test conditions. PMID- 12898630 TI - Easy access to (R,R)-3,4-dihydroxy-3,4-dihydrobenzoic acid with engineered strains of Escherichia coli. PMID- 12898631 TI - Direct visualization of DNA duplex formation on the surface of a giant liposome. PMID- 12898634 TI - Gene therapy for HIV/AIDS: the potential for a new therapeutic regimen. AB - Human Immunodeficiency Virus (HIV) is the etiologic agent of Acquired Immunodeficiency Syndrome (AIDS). HIV/AIDS is a disease that, compared with the not so distant past, is now better held in check by current antiretroviral drugs. However, it remains a disease not solved. Highly active antiretroviral therapy (HAART) generally uses two non-nucleoside and one nucleoside reverse transcriptase (RT) inhibitor or two non-nucleoside RT and one protease inhibitor. HAART is far more effective than the mono- or duo-therapy of the past, which used compounds like the nucleoside reverse transcriptase inhibitor AZT or two nucleoside reverse transcriptase inhibitors. However, even with the relatively potent drug cocktails that comprise HAART, there are the issues of (i). HIV escape mutants, (ii). an apparent need to take the drugs in an ongoing manner, and (iii). the drugs' side effects that are often severe. This review speaks to the potential addition to these potent regimens of another regimen, namely the genetic modification of target hematopoietic cells. Such a new treatment paradigm is conceptually attractive as it may yield the constant intracellular expression of an anti-HIV gene that acts to inhibit HIV replication and pathogenicity. A body of preclinical work exists showing the inhibition of HIV replication and decreased HIV pathogenicity by anti-HIV genetic agents. This preclinical work used hematopoietic cell lines and primary cells as the target tissue. More recently, several clinical trials have sought to test this concept in vivo. PMID- 12898635 TI - Lentivirally transduced dendritic cells as a tool for cancer immunotherapy. AB - BACKGROUND: Dendritic cells (DC) are the professional antigen-presenting cells of the immune system, fully equipped to prime naive T cells and thus essential components for cancer immunotherapy. METHODS: We tested the influence of several elements (cPPT, trip, WPRE, SIN) on the transduction efficiency of human DC. Human and murine DC were transduced with tNGFR-encoding lentiviruses to assess the effect of transduction on phenotype and function. Human DC were transduced with lentiviruses encoding huIi80MAGE-A3 and murine DC with huIi80tOVA to test antigen presentation. RESULTS: A self-inactivating (SIN) lentiviral vector containing the trip element was most efficient in transducing human DC. The transduction of DC with trip/SIN tNGFR encoding lentiviral vectors at MOI 15 resulted in stable gene expression in up to 94.6% (murine) and 88.2% (human) of the mature DC, without perturbing viability, phenotype and function. Human huIi80MAGE-A3-transduced DC were able to stimulate MAGE-A3-specific CD4(+) and CD8(+) T cell clones and could prime both MAGE-A3-specific CD4(+) and CD8(+) T cells in vitro. Murine huIi80tOVA-transduced DC were able to present OVA peptides in the context of MHC class I and class II in vitro and induced a strong OVA specific cytotoxic T lymphocyte response in vivo, that was protective against subsequent challenge with OVA-expressing tumor cells. CONCLUSIONS: We show that, using lentiviral vectors, efficient gene transfer in human and murine DC can be obtained and that these DC can elicit antigen-specific immune responses in vitro and in vivo. The composition of the transfer vector has a major impact on the transduction efficiency. PMID- 12898636 TI - Dendritic cells engineered to express the Flt3 ligand stimulate type I immune response, and induce enhanced cytoxic T and natural killer cell cytotoxicities and antitumor immunity. AB - BACKGROUND: Tumor antigen presentation by dendritic cells (DCs) to T cells in lymphoid organs is crucial for induction of antitumor immune responses. Fms-like tyrosine kinase 3 ligand (Flt3L) is a regulator of hematopoietic cell development. METHODS: To investigate the potential effect of Flt3L transgene expression on DC-based cancer vaccines, we constructed a recombinant adenovirus AdVFlt3L expressing Flt3L, transfected DCs with AdVFlt3L, and investigated the efficacy of antitumor immunity by vaccination of DC(Flt3L) engineered to express Flt3L transgene. RESULTS: Our data demonstrated that AdVFlt3L transfection up regulated the expression of cytokine IL-1beta and chemokines MIP-1alpha, MIP 1beta, IP-10, MCP-1 and MIP-2, and stimulated DC(Flt3L) cell proliferation in vitro and migration toward regional lymph nodes in vivo. Our data also demonstrated that vaccination of Mut1-pulsed DC(Flt3L) cells was able to stimulate (i). a type 1 immune response comprising CD4(+) Th1 and CD8(+) Tc1 activation and (ii). around 2- and 3-fold enhanced tumor-specific cytotoxic T lymphocyte (CTL) and non-specific NK responses (p < 0.05) than vaccination with similarly pulsed control virus-transfected and untransfected DCs, respectively. More importantly, vaccination of Mut1-pulsed DC(Flt3L) cells induced enhanced antitumor immunity in vivo, even against poorly immunogenic 3LL tumor cells. Vaccinations of Mut1-pulsed DCs, DC(pLpA) and DC(Flt3L) all protected mice from challenge of low dose (0.5 x 10(5)) tumor cells. However, only vaccination of the last one was able to protect 63% (6/8) mice from challenge of high dose (3 x 10(5)) 3LL tumor cells (p < 0.01). CONCLUSIONS: DCs engineered to secrete Flt3L may offer a new strategy in DC-based cancer vaccines. PMID- 12898637 TI - Proliferative activity and in vitro replication of HSV1716 in human metastatic brain tumours. AB - BACKGROUND: The neurotropic herpes simplex virus mutant HSV1716 lacks the gene encoding the virulence factor ICP34.5 and cannot replicate in non-dividing cells where proliferating cell nuclear antigen (PCNA) is not actively engaged in cellular DNA synthesis. In the brain, tumoral expression of PCNA therefore confers on it oncolytic specificity and may determine its efficacy. Three phase I trials in glioma patients and one in metastatic melanoma patients have established that HSV1716 is safe and replicates selectively in tumour tissue. Here we examine the in situ PCNA profiles of common human metastatic brain tumours and determine their in vitro permissivity for HSV1716 replication to ascertain their suitability for HSV1716 therapy. METHODS: Histological sections of tumour biopsies obtained from patients undergoing craniotomies were stained for PCNA expression. The replicative ability of HSV wild-type (17(+)) and mutant (1716) viruses was assessed in tissue cultures of the same tumour biopsies and in cancer cell lines by plaque assay. RESULTS: Biopsies of all 10 metastatic tumours (3 melanoma, 4 carcinoma and 3 adenocarcinoma) as well as 4 glioblastoma multiforme were positive for PCNA immunoexpression and supported the replication of HSV1716. The PCNA-positive cells in the metastatic tumours were distributed comparatively in larger and more contiguous areas than in glioblastoma (1.69 +/- 1.61 mm(2) vs. 0.73 +/- 0.77 mm(2)) and numbered 29.0 +/- 12.4 and 12.6 +/- 4.7%, respectively. CONCLUSIONS: The results show that human cerebral metastatic tumours have generally larger and more contiguous proliferative areas, support efficient HSV1716 replication, and are thus potential candidates for such oncolytic viral therapy. PMID- 12898638 TI - Phase I immunotherapy with a modified vaccinia virus (MVA) expressing human MUC1 as antigen-specific immunotherapy in patients with MUC1-positive advanced cancer. AB - BACKGROUND: The MUC1 protein is a highly glycosylated mucin normally found at the apical surface of mucin-secreting epithelial cells in many types of tissues. MUC1 is expressed, but heavily underglycosylated, in different human tumors. TG4010 is a viral suspension of a recombinant vaccinia vector (MVA) containing DNA sequences coding for the human MUC1 antigen and interleukin-2 (IL-2). This product was developed for use as a vaccine in cancer patients whose tumors express the MUC1 antigen. The objective of the present study was to determine the safety of the product and to define the dose of TG4010 to be used in further clinical trials. MATERIALS AND METHODS: Thirteen patients with different solid tumors were treated by repeated intramuscular injection with increasing doses of TG4010 in two separate phase I studies, one in Europe (Basel-CR) and one in the United States (UCLA-RF): a total of 6 patients were treated at a dose of 5 x 10(6) pfu, 3 patients at 5 x 10(7) pfu, and 4 patients at 10(8) pfu. Safety, efficacy, and different immunological tests were the endpoints of the study. RESULTS: Tolerance of TG4010 was excellent, and side effects mainly consisted of injection site pain and influenza-like symptoms. There was no apparent detrimental effect of repeated injections of the vaccinia virus. Four of thirteen evaluable patients showed stabilization of their disease for 6 to 9 months. One lung cancer patient who was initially progressing after the first injections later showed a marked decrease in the size of his metastases that lasted for 14 months. Some T cell proliferative immune responses were seen in five patients. CONCLUSIONS: The administration of TG4010 was generally well tolerated in patients with metastatic tumors, and transient disease stabilization was observed in several patients, warranting further clinical studies with the product. PMID- 12898639 TI - Low molecular weight protamine as an efficient and nontoxic gene carrier: in vitro study. AB - BACKGROUND: The structural similarity between low molecular weight protamine (LMWP), prepared by enzymatic digestion of protamine, and HIV-TAT protein transduction peptide suggested the feasibility of LMWP as an efficient carrier for delivering therapeutic genes while alleviating the cytotoxicity of currently employed gene carriers. METHODS: LMWP was prepared by enzymatic digestion of protamine with thermolysine. The prepared LMWP peptide and TAT peptide, as well as their complexes with plasmid DNA (pDNA), were examined for cellular uptake behaviors by using confocal microscopy and flow cytometry. The complexation of pDNA and LMWP was monitored by gel retardation test as well as size and zeta potential measurements, and was then further assessed by DNase I protection assay. The transfection efficiency of pDNA/LMWP was examined by varying the pDNA content and charge ratio in the complex, and then compared with that of pDNA/PEI. Cytotoxicity induced by pDNA/LMWP and pDNA/PEI was also examined. RESULTS: Prepared LMWP showed similar transcellular localization behavior and kinetics to those of TAT, and efficiently transferred the pDNA into nucleus and cytoplasm in a short time period. The size and zeta potential of the pDNA/LMWP complex were 120 nm and 30 mV, respectively, which were adequately suitable for cellular uptake. After forming the complex, LMWP appeared to effectively protect pDNA against DNase I attack. The pDNA/LMWP complex showed significantly enhanced gene transfer than both naked pDNA and the pDNA/PEI complex, while exhibiting a markedly reduced cytotoxicity than that of the pDNA/PEI complex. CONCLUSIONS: The present study suggested that LMWP could be a useful and safe tool for enhancing delivery of bioactive molecules and therapeutic DNA products into cells when applied in gene therapy. PMID- 12898640 TI - Increased non-viral gene transfer levels in mice by concentration of cationic lipid DNA complexes formed under optimized conditions. AB - BACKGROUND: A number of non-viral gene transfer reagents including cationic lipid DNA complexes (LDC) have been developed and were improved by changing the ratio of their components. To determine the effect of other parameters during complex formation affecting the efficacy of LDC, the conditions during complexation were varied without changing the ratios of the components. METHODS: LDC were formed at fixed ratios of an equimolar mixture of DOTAP and cholesterol to DNA to the condensing agent protamine sulfate according to different protocols at varying final concentrations. The influence of these parameters on transfection efficiency and physical properties of the complexes was determined. RESULTS: Changing the order of addition of compounds during complex formation affected the size distribution, the charge of the LDC, the interaction between the lipids and the accessibility of the DNA. At fixed ratios of the components, higher transfection efficiencies were observed with more condensed LDC. Complexation in higher volumes increased transduction efficiency of the complexes after intravenous inoculation. Due to restrictions on the injectable volume, the LDC were formed in the optimal volume and subsequently concentrated by ultrafiltration. The concentrated complexes maintained transduction efficiency and up to 60-fold higher in vivo transduction levels were obtained. CONCLUSIONS: In addition to the ratio of the components of cationic lipid DNA complexes, the final concentration and the order of addition of compounds during complex formation are critical for high transduction efficiency. Concentration of LDC formed under optimal conditions can be used to further increase in vivo gene transfer levels. PMID- 12898641 TI - Development of a dual-luciferase fusion gene as a sensitive marker for site directed DNA repair strategies. AB - BACKGROUND: Several novel techniques have been developed recently for the site specific repair of DNA as an approach to gene therapy. Correction efficiencies as high as 40% have been reported, well within the range of therapeutic impact for a number of genetic diseases. Unfortunately, many of the model systems in which these methods have been employed typically target genes that are not well suited for analyzing the various techniques. METHODS: To address this, we have constructed and characterized a dual-luciferase fusion gene as a sensitive marker for optimizing repair strategies. The genes encoding two distinct luciferase proteins were fused so that expression of one luciferase necessitated expression of the other. Engineering a stop codon in the downstream luciferase gene created an ideal tool to study the efficiency of various site-directed DNA repair techniques as one luciferase can act as an internal control while the other is targeted for correction. RESULTS: Fusing two luciferase genes resulted in a single protein that produces two bioluminescent activities in a constant ratio. The utility of this system as a target for site-directed DNA repair research was demonstrated using two of the recently developed gene repair techniques, small fragment homologous replacement and oligonucleotide-mediated repair, to mediate correction and by the ability to detect repair efficiencies of less than 5 x 10( 6) (<1 event in 200000). CONCLUSIONS: The ability to rapidly and accurately quantify the amount of correction using the dual-luciferase fusion system will allow the comparison and evaluation of the many factors involved in successful gene repair and lead to the optimization of these techniques, both in cell culture and in whole animals. PMID- 12898643 TI - In vitro human phase I metabolism of xenobiotics I: pesticides and related compounds used in agriculture and public health, May 2003. AB - This is the first revision of a database covering human phase I enzymes and their isoforms that metabolize pesticides and related compounds. The original version included enzymes that metabolize chloroacetamide and triazine herbicides, and organophosphorus insecticides. This revision also includes carbamate, nicotinoid, and pyrethroid insecticides and insect repellents. PMID- 12898644 TI - Influence of culture time on the expression of drug-metabolizing enzymes in primary human hepatocytes and hepatoma cell line HepG2. AB - Primary cultures of human hepatocytes and hepatoma cell line HepG2 are frequently used to evaluate the hepatic disposition of drugs and other xenobiotics. To check the variability of the expression of drug-metabolizing enzymes in these in vitro models, expression of genes coding for several cytochrome P450 isoforms and phase II enzymes was quantified during culture time by real-time RT-PCR. Gene expression was determined daily for primary hepatocytes maintained in a sandwich culture over 1 week and for HepG2, during the first 10 passages. In primary hepatocytes characteristic expression trends were observed which could be abstracted into three major classes of time curves. Genes of the first and the second class had an expression maximum around day 6 and day 4 in culture, respectively. The third class of genes had two expression peaks: at day 1 and 5 in culture. Surprisingly, also the cell line HepG2 showed significant expression changes during passages. For example, gene expression of cytochrome 1A1 varied 8 fold, that of cytochrome 2B6 30-fold, and that of NADP-quinone reductase 1 more than 200-fold within the first 10 passages. In conclusion, neither primary hepatocytes nor HepG2 cell line display a model for constant expression of drug metabolizing enzymes. PMID- 12898645 TI - Diabetes and mitochondrial bioenergetics: alterations with age. AB - Several studies have been carried out to evaluate the alterations in mitochondrial functions of diabetic rats. However, some of the results reported are controversial, since experimental conditions, such as aging, and/or strain of animals used were different. The purpose of this study was to evaluate the metabolic changes in liver mitochondria, both in the presence of severe hyperglycaemia (STZ-treated rats) and mild hyperglycaemia (Goto-Kakizaki (GK) rats). Moreover, metabolic alterations were evaluated both at initial and at advanced states of the disease. We observed that both models of type 1 and type 2 diabetes presented alterations on respiratory chain activity. Because of continual severe hyperglycaemia, 9 weeks after the induction of diabetes, the respiratory function declined in STZ-treated rats, as observed by membrane potential and respiratory ratios (RCR, P/O, and FCCP-stimulated respiration) assessment. In contrast, GK rats of 6 months age presented increased respiratory ratios. To localize which respiratory complexes are affected by diabetes, enzymatic respiratory chain activities were evaluated. We observed that succinate dehydrogenase and cytochrome c oxidase activities were significantly augmented both in STZ-treated rats and GK rats of 6 months age. Moreover, H(+)-ATPase activity was also significantly increased in STZ-treated rats with 3 weeks of diabetes and in GK rats of 6 months age as compared to controls. Therefore, these results clearly suggest that both animal models of diabetes present some metabolic adjustments in order to circumvent the deleterious effects promoted by the high glucose levels typical of the disease. PMID- 12898646 TI - Argemone oil induced cellular damage in the reproductive tissues of transgenic Drosophila melanogaster: protective role of 70 kDa heat shock protein. AB - We explored the reproductive toxicity of argemone oil and its principal alkaloid fraction in transgenic Drosophila melanogaster (hsp70-lacZ) Bg(9). The toxicity of argemone oil has been attributed to two of its physiologically active benzophenanthridine alkaloids, sanguinarine and dihydrosanguinarine. Freshly eclosed first instar larvae of transgenic Drosophila melanogaster were transferred to different concentrations of argemone oil and its alkaloid fraction contaminated food. Virgin flies that eclosed from the contaminated food were pair mated to look into the effect on reproduction. The study was further extended by investigating hsp70 expression and tissue damage in larval gonads, genital discs, and reproductive organs of adult fly. Our results showed that argemone oil was more cytotoxic than its principal alkaloid fraction. Moreover, it was the male fly that was more affected compared to its opposite number. The accessory glands of male reproductive system of the fly, which did not express hsp70, exhibited severe damage as evidenced by Trypan blue staining. This prompted us to explore the ultrastructural morphology of the gland, which showed acute signs of necrosis in both the cell types as evident by necrotic nuclei, higher vacuolization, and disorganized endoplasmic reticulum, decrease in the number of Golgi vesicles and disorganized, loosely packed filamentous structures in the lumen of the accessory gland, at the higher concentrations of the adulterant. The study showed the reproductive toxicity of argemone oil and its alkaloid fraction in transgenic Drosophila melanogaster and further confirmed the cytoprotective role of hsp70. PMID- 12898647 TI - Evidence for the existence of a common ancestor of scorpion toxins affecting ion channels. AB - All scorpion toxins from different 30 species are simply reviewed. A new classification system of scorpion toxins is first proposed: scorpion toxins are classified into three families (long-chain scorpion toxins with 4 disulfide bridges, short-chain scorpion toxins with 3 disulfide bridges, and intermediate type scorpion toxins with 3 or 4 disulfide bridges). Intermediate-type scorpion toxins provide a strong proof for the conclusion that channel toxins from scorpion venoms evolve from a common ancestor. Common organization of precursor nucleotides and genomic sequence, similar 3-dimensional structure, and the existence of intermediate type scorpion toxins and functionally intercrossing scorpion toxins show that all scorpion toxins affecting ion channels evolve from the common ancestor, which produce millions of scorpion toxins with function diversity. PMID- 12898648 TI - Binding of 3-hydroxybenzo[a]pyrene to bovine hemoglobin and albumin. AB - Previous studies examined the bioavailability and first-pass biotransformation of 3-hydroxy[(3)H]benzo[a]pyrene ([(3)H]-3-OHBaP) in an isolated perfused catfish intestinal model. This work showed that 3-OHBaP, or a metabolite formed in intestine, bound covalently to blood protein. In this study, the blood adducts were characterized in vitro by incubating bovine ferric hemoglobin or albumin with [(3)H]-3OHBaP under various conditions. Incubation of 2 microM [(3)H]-3 OHBaP with hemoglobin for 1 h resulted in 7.49 pmol bound/mg protein, while albumin binding was 1.37 pmol/mg protein. Mild acid hydrolysis released only 5% of the radioactivity from 3-OHBaP-hemoglobin adducts. After gel filtration, the 3 OHBaP-hemoglobin adducts were examined by HPLC analysis. A single peak of radioactivity was detected at the same retention time as the heme component of hemoglobin. Unbound 3-OHBaP was oxidized to BaP-3,6-dione during incubation with ferric hemoglobin. Treatment of hemoglobin with ascorbic acid decreased the formation of hemoglobin adducts by 33%, while hydrogen peroxide treatment increased adduct formation by 44%. Incubation of [(3)H]-BaP-3-beta-D-glucuronide (BaP-3G) with hemoglobin and beta-glucuronidase resulted in greater binding to hemoglobin than incubation with [(3)H]-3-OHBaP alone. The hemoglobin adduct obtained from [(3)H]-BaP-3G also co-migrated with heme. These results indicate that an oxidative process is involved in formation of the heme adduct and that 3 OHBaP or BaP-3G might be a precursor of the bound metabolite. PMID- 12898649 TI - Direct tissue analysis using matrix-assisted laser desorption/ionization mass spectrometry: practical aspects of sample preparation. AB - Practical guidelines for the preparation of tissue sections for direct analysis by matrix-assisted laser desorption/ionization (MALDI) mass spectrometry are presented. Techniques for proper sample handling including tissue storage, sectioning and mounting are described. Emphasis is placed on optimizing matrix parameters such as the type of matrix molecule used, matrix concentration, and solvent composition. Several different techniques for matrix application are illustrated. Optimal instrument parameters and the necessity for advanced data analysis approaches with regards to direct tissue analysis are also discussed. PMID- 12898650 TI - Derivatization reaction of the mycotoxin moniliformin with 1,2-diamino-4,5 dichlorobenzene: structure elucidation of an unexpected reaction product by liquid chromatography/tandem mass spectrometry and liquid chromatography/nuclear magnetic resonance spectroscopy. AB - The derivatization reaction of the mycotoxin moniliformin with 1,2-diamino-4,5 dichlorobenzene was previously introduced to improve distinctly the sensitivity of an assay applying high-performance liquid chromatography prior to fluorescence detection. In the course of the implementation of this derivatization approach into a liquid chromatographic/mass spectrometric method, an unexpected derivatization product has now been discovered by mass spectrometry. In order to elucidate its structure, detailed investigations with liquid chromatography/tandem mass spectrometry and liquid chromatography coupled on-line with NMR spectroscopy were performed. These studies give evidence for a heterocyclic structure that has been formed by the loss of one water and one carbon monoxide molecule. A reasonable mechanism for this derivatization reaction is proposed. PMID- 12898651 TI - Internal residue loss produced by rearrangement of a novel cationic glycosphingolipid, glyceroplasmalopsychosine, in collision-induced dissociation. AB - A novel plasmal conjugate of galactosylsphingosine (psychosine), Gro1(3)-O plasmal-O-6Galbeta-sphingosine (glyceroplasmalopsychosine), was analyzed by electrospray ionization and liquid secondary ion mass spectrometry with low- or high-energy collision-induced dissociation (CID). In the product ion spectra of the [M + H](+) ions, [M + H - glycerol](+) ions arising from the loss of a glycerol were predominant. Unexpectedly, CID of the [M + H - glycerol](+) ion produced an outstanding ion, [(M + H - glycerol) - Hex](+), which required the loss of the galactose from inside the molecule. This ion was greatly reduced in the spectra of N,N-dimethyl derivatives, indicating that the [(M + H - glycerol) Hex](+) ion is formed from an intramolecular rearrangement with migration of the plasmal residue to the free amino group of sphingosine. It would be expected that the rearrangement occurs simultaneously with the elimination of glycerol or a rearranged [M + H](+) ion leads to the elimination of glycerol, to form a Schiff base-type [M + H - glycerol](+) ion, from which the terminal galactose could be removed by the normal mechanism of glycosidic cleavage. On the other hand, the [M + Na - glycerol](+) ion derived from the sodiated molecule did not produce an ion corresponding to the rearrangement reaction, possibly owing to a higher stability of the sodiated ions against conformational changes. PMID- 12898652 TI - Studies on the non-covalent complexes between oleanolic acid and cyclodextrins using electrospray ionization tandem mass spectrometry. AB - Non-covalent inclusion complexes formed between an anti-inflammatory drug, oleanolic acid (OA), and alpha-, beta- and gamma-cyclodextrins (CDs) were investigated by means of solubility studies and electrospray ionization tandem mass spectrometry (ESI-MS(n)). The order of calculated association constants (K(1 : 1)) of complexes between OA and different CDs in solution is in good agreement with the order of their relative peak intensities and the relative CID energies of the complexes under the same ESI-MS(n) conditions. These results indicate a direct correlation between the behaviors of solution- and gas-phase complexes. ESI-MS can thus be used to evaluate solution-phase non-covalent complexes successfully. The experimental results show that the most stable 1 : 1 inclusion complexes between three CDs and OA can be formed, but 2 : 1 CD-OA complexes can be formed with beta- and gamma-CDs. Multi-component complexes of alpha-CD-OA-beta CD (1 : 1 : 1), alpha-CD-OA-gamma-CD (1 : 1 : 1) and beta-CD-OA-gamma-CD (1 : 1 : 1) were found in equimolar CD mixtures with excess OA. The formation of 2 : 1 and multi-component 1 : 1 : 1 non-covalent CD-OA complexes indicates that beta- and gamma-CD are able to form sandwich-type inclusion non-covalent complexes with OA. The above results can be partly supported by the relative sizes of OA and CD cavities by molecular modeling calculations. All the complexes allow the detection of gaseous deprotonated CD-OA complexes in the negative ion mode at high abundances. The relative stabilities of the CDs-OA inclusion complexes in the gas phase can be evaluated from the relative CID energies in the ion trap (alpha-CD-OA < beta-CD-OA < gamma-CD-OA) in the negative ion mode. PMID- 12898653 TI - A sensitive and selective liquid chromatographic/electrospray ionization tandem mass spectrometric assay for the simultaneous quantification of alpha-,beta arteether and its metabolite dihydroartemisinin in plasma, useful for pharmacokinetic studies. AB - A sensitive, selective, specific and rapid liquid chromatographic/electrospray ionization tandem mass spectrometric assay method was developed and validated for the simultaneous quantitation of alpha-,beta-arteether (alpha-,beta-AE) and its metabolite alpha-dihydroartemisinin (DHA) in monkey plasma using the propyl ether analogue of beta-arteether (PE) as an internal standard. The method involves a simple two-step liquid-liquid extraction with hexane. The analytes were chromatographed on a C(18) reversed-phase chromatographic column by isocratic elution with methanol-ammonium acetate buffer (pH 4) (92 : 8, v/v) and analysed by mass spectrometry in the multiple reaction monitoring mode. The chromatographic run time was 7 min and the weighted (1/x(2)) calibration curves were linear over the range 0.78-200 ng ml(-1). The method was validated in terms of accuracy, precision, absolute recovery, freeze-thaw stability, bench-top stability and re-injection reproducibility. The limit of detection and lower limit of quantification in monkey plasma were 0.39 and 0.78 ng ml(-1) respectively for all the analytes. The intra- and inter-batch precision and accuracy were found to be well within acceptable limits (<15%). All three analytes were stable even after three freeze-thaw cycles (deviation < 15%). The average absolute recoveries of alpha-,beta-AE, DHA and PE, used as an internal standard, from spiked plasma samples were 85.85 +/- 6.56, 70.10 +/- 7.06, 54.37 +/- 3.39 and 93.90 +/- 6.9%, respectively. The assay method described here could be applied to study the pharmacokinetics of alpha-,beta-AE and DHA in rhesus monkeys. PMID- 12898654 TI - High-sensitivity quantitation of cabergoline and pergolide using a triple quadrupole mass spectrometer with enhanced mass-resolution capabilities. AB - Quantitative analysis of pharmaceuticals with low systemic plasma levels requires the utmost in sensitivity and selectivity from the analytical method used. A recently introduced triple-quadrupole mass spectrometer with unique enhanced mass resolution capability was evaluated in the analysis of two such drugs, cabergoline and pergolide, in plasma. Liquid chromatographic/electrospray ionization selected reaction monitoring determination of cabergoline in plasma at unit mass-resolution demonstrated improved sensitivity (50 fg on-column), coupled with suitable accuracy and precision over a broad linear dynamic range covering five orders of magnitude (50 fg to 5 ng on-column). Liquid chromatographic/atmospheric pressure chemical ionization selective reaction monitoring determination of pergolide in plasma also attained a high level of sensitivity (500 fg on-column) at unit mass-resolution, with accuracy and precision values well within pharmaceutical industry standards. Again, a linear dynamic range covering five orders of magnitude (500 fg to 50 ng on-column) was achieved for the assay. Utility of the enhanced mass-resolution feature of the triple-quadrupole mass spectrometer in the determination of pergolide resulted in an improvement in analyte sensitivity (250 fg on-column) and linear dynamic range (250 fg to 50 ng on-column). PMID- 12898655 TI - Characterization of alkylacyl, alk-1-enylacyl and lyso subclasses of glycerophosphocholine by tandem quadrupole mass spectrometry with electrospray ionization. AB - Positive ion tandem quadrupole mass spectrometric methods for structural characterization of the subclasses of sn-glycero-3-phosphocholine (PC), including alkylacyl- and alk-1-enylacylphosphocholine and lysophosphatidylcholine (LPC), are described. Following collisionally activated dissociation, the [M + Li](+) ions generated by electrospray ionization yield abundant informative fragment ions that permit structural determination, and distinction of regioisomers among lysophosphatidylcholine can be easily achieved. In contrast, structurally informative ions arising from [M + H](+) or [M + Na](+) ions are less prominent. The most abundant ion observed in the product-ion spectra of the [M + Li](+) ions of plasmenyl- and plasmanyl-PC and of LPC arises from loss of N(CH(3))(3) ([M + Li - 59](+)). This feature permits their distinction from a product-ion spectrum arising from a diacylphosphatidylcholine, in which the [M + Li - 183](+) ion reflecting loss of phosphocholine is the most prominent. Examples for identification of various subclasses of PC in biological extracts by tandem mass spectrometry applying various constant neutral loss scannings are also shown. PMID- 12898656 TI - Mass spectrometric methods for the analysis of chlorinated and nitrated isoflavonoids: a novel class of biological metabolites. AB - Electrospray ionization combined with tandem mass spectrometry was applied to a study of some representative chlorinated and nitrated isoflavones-potential metabolites of isoflavones in inflammatory cells. Upon collision-induced dissociation of deprotonated [M - H](-) ions of these compounds, a number of structurally characteristic product ions were produced. The product ion analysis of 3'- and 8-chlorodaidzein in the tandom mass spectra led to ready differentiation of these isomers. 3-Nitro derivatives of both genistein and daidzein have product ions due to the losses of HNO(2) and two OH groups. Chlorinated derivatives of isoflavones were detected in cell-based experiments and their structures were proposed by comparing the tandem mass spectra of their product ions with those of standards. This work provides a suitable analytical basis to aid the characterization of chlorinated and nitrated metabolites in studies in vivo and in vitro. PMID- 12898657 TI - Two-laser infrared and ultraviolet matrix-assisted laser desorption/ionization. AB - Matrix-assisted laser desorption/ionization (MALDI) was performed using two pulsed lasers with wavelengths in the IR and UV regions. A 10.6 micro m pulsed CO(2) laser was used to irradiate a MALDI target, followed after an adjustable delay by a 337 nm pulsed nitrogen laser. The sample consisted of a 2,5 dihydroxybenzoic acid matrix and bovine insulin guest molecule. The pulse energy for both of the lasers was adjusted so that the ion of interest, either the matrix or guest ion, was not produced by either of the lasers alone. The delay time for maximum ion yield occurs at 1 micro s for matrix and guest ions and the signal decayed to zero in approximately 400 micro s. A mechanism is presented for enhanced UV MALDI ion yield following the IR laser pulse based on transient heating. PMID- 12898658 TI - Energetics of an intracluster beta-elimination process driven by acetate anions. The case of a Fmoc-protected peptide investigated by high-resolution electrospray ionization tandem mass spectrometry. PMID- 12898659 TI - Current literature in mass spectrometry. PMID- 12898660 TI - Construction of the contingent valuation market in health care: a critical assessment. AB - Contingent valuation (CV) has been criticised for being too hypothetical, with expressed values bearing little relation to actual values. The magnitude of this divergence, however, depends upon how realistic and believable the contingent market is. This paper presents an overview of five key aspects in the construction of the contingent market: (i) scenario development and presentation; (ii) payment vehicle; (iii) expression of risk; (iv) time period of valuation; and (v) survey administration. CV studies in health care since 1985, totalling 111, are critically reviewed with respect to these five aspects. It is concluded that CV studies in health care have performed poorly in the construction, specification and presentation of the contingent market, and that there has been little, if any, improvement in this respect over the last 15 years. Suggestions are made concerning why this may be the case, and how the construction of the contingent market may be improved in future. PMID- 12898661 TI - The missing link: on the line between C and E. AB - In this paper it is argued that the separation of elements associated with the time spent by the patient is not conducted in a consistent way. This is the case for income (for which there at least has been some attention) and for other time elements like lost unpaid work, leisure and role-functioning. The use of general rather than specific preferences in health state assessments makes the separation of time-elements into costs and effects difficult. While costs are calculated specifically for the patient group under study, effects are normally derived from preferences in the general public. The characteristics of these two groups in terms of (the opportunity of) spending time on activities need not coincide. The use of specific time-group valuations of health states may be a good alternative to using general health state valuations. PMID- 12898662 TI - The value of the change in health in Sweden 1980/81 to 1996/97. AB - The study aimed to estimate the value of the change in health in Sweden 1980/81 to 1996/97. Quality-adjusted life years (QALYs) for men and women at specific ages were estimated for 1980/81, 1988/89 and 1996/97, by combining survival rates and health state scores. Data from the Swedish Survey of Living Conditions (n = 39,966) were used to estimate age-specific health state scores. Responses to selected survey questions were mapped into the EQ-5D measure, using the UK EQ-5D index tariff to derive health state scores. The monetary value of a QALY was assumed to be 100,000 dollars. Life expectancy for infants increased by 3.68 years for males and 2.70 years for females between 1980/81 and 1996/97. Average health status decreased in younger age groups whereas it increased in older age groups. Expected QALYs for infants increased by 2.64 for males and 0.54 for females. With 3% discounting the gain was 0.11 QALYs (11,000 dollars) among males and a loss by 0.58 QALYs (58,000 dollars) among females. The corresponding gain in discounted QALYs for a 75-year-old was 1.15 (115,000 dollars) and 0.80 (80,000 dollars), respectively. It is concluded that older persons have experienced considerable health gains whereas the health gains have been small or non existent for younger women. PMID- 12898663 TI - The TTO method and procedural invariance. AB - In a pilot study we investigate whether the inferences we draw about people's preferences towards health care treatments are altered if we vary the procedure that is used to elicit these preferences. In a conventional time trade-off (TTO) question, respondents express their preferences towards treatment by comparing a period of ill-health with a shorter period in a higher quality of life. In our less conventional TTO question, we vary the procedure by asking respondents their preferences towards treatment by comparing a period of ill-health with a longer period in a lower quality of life. The quantitative data are equivocal about whether preferences for treatment differ between the conventional and unconventional questions. The qualitative data support the notion of contrasting issues in the questions that involve prolonging time in a lower quality of life and appear to account for a failure to find quantitative differences in all of the questions. PMID- 12898664 TI - Hospital efficiency targets. AB - Cost-efficiency targets, used to encourage downward pressure on hospital unit costs, have been employed within the UK NHS for many years. There has been considerable speculation that these targets create incentives to reduce beds and increase occupancy rates at the expense of holding spare capacity to accommodate fluctuations in emergency admissions. This research used panel data for the period 1994/1995-1999/2000, supplemented by a series of semi-structured interviews, to explore the strategies Trusts employ to reduce unit costs. No relationship could be found between published targets and changes in unit costs, nor that targets were successful in reducing the dispersion of unit costs over time. Interviews revealed that efficiency gains required of Trusts, usually dictated by the local health economy, often bore no correspondence to the national or regional published targets. Results further indicated that contrary to prior speculation, Trusts divide into two distinct groups, those with high occupancy rates and those with a high proportion of free beds to accommodate emergencies, with Trust characteristics displaying stability over time. A pressing need for future work is the development of measures to encourage efficiency that take account of quality improvement. PMID- 12898665 TI - Public spending on health care and the poor. AB - This paper uses new cross-country data to assess the relationship between public spending on health care and the health status of the poor. Data are drawn from two sources: (i) existing data on health status by income quintile tabulated from demographic health surveys in 44 countries; and (ii) our estimates of the health status of the poor in over 70 countries drawn from a new technique in decomposing social indicators. Our estimates confirm that the poor have significantly worse health status than the nonpoor and the regression results provide new evidence that public spending on health care matters more to them. However, the results suggest that increased public spending alone will not be sufficient to significantly improve health status. PMID- 12898666 TI - Methodological issues raised by preference-based approaches to measuring the health status of children. AB - This paper identifies and discusses a number of methodological issues that require consideration when applying preference-based approaches to the measurement of the health status of children. It is argued that the decision about which dimensions of health status to incorporate into health state descriptions or classifications should depend, in part, upon whether the measure will be used to inform resource allocation within or across age groups. In addition, the paper identifies and discusses a number of methodological issues surrounding the appropriate respondents for descriptions and valuations of health status in different contexts; potential sources of bias in the description and valuation processes; and the psychometric integrity of alternative measurement approaches. PMID- 12898667 TI - Solid supports for microarray immunoassays. AB - Stimulated by the achievements of the first phase in genomics and the resulting need of assigning functions to the acquired sequence information, novel formats of immunoassays are being developed for high-throughput multi-analyte studies. In principle, they are similar in nature to the microarray assays already established at the level of nucleic acids. However, the biochemical diversity and the sheer number of proteins are such that an equivalent analysis is much more complex and thus difficult to accomplish. The wide range of protein concentration complicates matters further. Performing microarray immunoassays already represents a challenge at the level of preparing a working chip surface. Arrays have been produced on filter supports, in microtiter plate wells and on glass slides, the last two usually coated with one-, two- or three-dimensionally structured surface modifications. The usefulness and suitability of all these support media for the construction and application of antibody microarrays are reviewed in this manuscript in terms of the different kinds of immunoassay and the various detection procedures. Additionally, the employment of microarrays containing alternative sensor molecules is discussed in this context. The sensitivity of microspot immunoassays predicted by the current analyte theory is not yet a reality, indicating the extent of both the technology's potential and the size of the task still ahead. PMID- 12898668 TI - Long-range and short-range mechanisms of hydrophobic attraction and hydrophilic repulsion in specific and aspecific interactions. AB - Among the three different non-covalent forces acting in aqueous media, i.e. Lifshitz-van der Waals (LW), Lewis acid-base (AB) and electrical double layer (EL) forces, the AB forces or electron-acceptor/electron-donor interactions are quantitatively by far the predominant ones. A subset of the AB forces acting in water causes the hydrophobic effect, which is the attraction caused by the hydrogen-bonding (AB) free energy of cohesion between the water molecules which surround all apolar as well as polar molecules and particles when they are immersed in water. As the polar energy of cohesion among water molecules is an innate property of water, the hydrophobic attraction (due to the hydrophobic effect) is unavoidably always present in aqueous media and has a value of DeltaG(hydrophobic) = -102 mJ/m(2), at 20 degrees C, being equal to the AB free energy of cohesion between the water molecules at that temperature. The strong underlying hydrophobic attraction due to this effect can, however, be surmounted by very hydrophilic molecules and particles that attract water molecules more strongly than the free energy of attraction of these molecules or particles for one another, plus the hydrogen-bonding free energy of cohesion between the water molecules, thus resulting in a net non-electrical double layer repulsion. Each of the three non-covalent forces, LW, AB or EL, any of which can be independently attractive or repulsive, decays, dependent on the circumstances, as a function of distance according to different rules. These rules, following an extended DLVO (XDLVO) approach, are given, as well as the measurement methods for the LW, AB and EL surface thermodynamic properties, determined at "contact". The implications of the resulting hydrophobic attractive and hydrophilic repulsive free energies, as a function of distance, are discussed with respect to specific and aspecific interactions in biological systems. The discussion furnishes a description of the manner by which shorter-range specific attractions can surmount the usually much stronger long-range aspecific repulsion, and ends with examples of in vitro and in vivo effects of hydrophilization of biopolymers, particles or surfaces by linkage with polyethylene oxide (PEO; also called polyethylene glycol, PEG). PMID- 12898669 TI - Molecular electrostatic potential as a factor of drug-receptor recognition. AB - When a drug molecule approaches a non-specific acceptor both molecules are in electrostatic fields of equal sign which prevents drug-acceptor complex formation. At the same time, the drug-acceptor system does not achieve the thermodynamic global minimum. Otherwise, when a certain drug interacts with its specific receptor, mutual compensation of their molecular electrostatic potentials takes place. Then separate atoms and groups of the drug molecule can bind to the receptor. We show that the fundamental role of molecular electrostatic potential in the process of drug-receptor recognition consists in fast correction of errors. PMID- 12898670 TI - Analysis of the pH-dependencies of the association and dissociation kinetics of HIV-1 protease inhibitors. AB - The kinetic constants for the interactions between HIV-1 protease and a selection of inhibitors were determined at different pH-values using a biosensor based interaction assay. Since this technique does not involve a substrate, it was possible to determine the pH-dependencies of the association and dissociation rates of an inhibitor, without the complication of a pH-dependent enzyme substrate/product equilibrium. The importance of these interactions was evaluated by correlating the free energy changes upon association and dissociation of inhibitors with the predicted change in electrostatic properties of the interacting groups as a result of altered pH. It was found that the kinetic parameters varied with pH in a unique manner for all inhibitors, demonstrating that the kinetic features were associated with the specific structure of each inhibitor. Association and dissociation had different pH-profiles, indicating that the two processes proceeded by different pathways/mechanisms. The energy barrier for dissociation of the enzyme-indinavir complex increased with pH from 4.1 to 7.4, while it was generally reduced for the other inhibitors as the pH was increased from 5.1 to 7.4. The pH-dependent interactions involved in the recognition/binding of inhibitors and in the stabilization of the complex were identified by analysing three-dimensional structures of enzyme-inhibitor complexes. The interaction between the pyridine nitrogen of indinavir with Arg-8 was hypothesized to be responsible for the unique pH-dependency of indinavir. The analysis revealed features of interactions that are significant for understanding enzyme function and for optimization of new drug leads. It also highlighted the importance of environmental conditions on interactions. PMID- 12898671 TI - Directional shape complementarity at the protein-DNA interface. AB - Nature utilizes various styles of architecture for DNA-binding proteins to recognize diverse DNA sequences, a process facilitated by a complementary surface between protein and DNA. However, the extent and ways this 'shape complementarity' occurs at the protein-DNA interface have yet to be characterized. Here, by analyzing a set of diverse protein-DNA complexes of known three-dimensional structures, we investigated whether the normal vectors of a protein surface at the interface exhibited any relationship with DNA conformation. Generally, the normal vectors of a DNA-contacting protein surface distinctly preferred certain angles, enabling them to align with certain axes characterizing the conformation of DNA. Thus, a new geometric property of DNA binding protein is demonstrated, i.e. the "shape complementarity" of protein-DNA recognition clearly bears the property of "directionality". PMID- 12898673 TI - Intramolecular dimers: a new design strategy for fluorescence-quenched probes. AB - Fluorogenic probes dual-labeled with reporter and quencher dyes use a change in fluorescence to monitor biochemical events (e.g., substrate binding or enzyme digestion). Such events change the reporter-quencher distance, which affects fluorescence. Recently, it is has been shown that static quenching through intramolecular dimers is an important mechanism that can sometimes be more efficient than Forster resonance energy transfer (FRET). PMID- 12898674 TI - From supramolecular block copolymers to advanced nano-objects. AB - The formation of asymmetric bis-complexes, based on terpyridine ligands and ruthenium ions, is described as a powerful tool for the self-assembly of polymer blocks end-functionalized with terpyridine units. This is illustrated in this contribution for the synthesis of amphiphilic metallo-supramolecular block copolymers, which are further used to produce aqueous micelles. Finally, the reversibility of the supramolecular bond opens new avenues for the preparation and manipulation of these nano-objects. PMID- 12898675 TI - Synthesis and solid-state organization of coil-ring-coil block copolymers. AB - Shape-persistent macrocycles based on the phenyl-ethynyl-butadienyl backbone containing two extraannular hydroxyl groups were prepared by the oxidative coupling of the appropriate phenylethynyl oligomers. Carbodiimide-directed coupling with independently synthesized polystyrene carboxylic acid oligomers led to ABA coil-ring-coil block copolymers in which the central macrocycle serves as rigid and the polystyrene oligomers as flexible elements. Depending on the size of the coil blocks, these structures aggregate in cyclohexane into supramolecular hollow cylindrical brushes in which the rigid core is surrounded by the flexible matrix. However, in the solid state it is not possible to identify a morphology in which isolated channels based on aggregated macrocycles are embedded in a matrix of polystyrene. Detailed X-ray and electron diffraction studies on samples prepared from a solution in cyclohexane under equilibrium conditions show that the material adopts a lamellar morphology in the solid state in which columns of macrocycles are aggregated into layers which are separated by polystyrene. PMID- 12898676 TI - Characteristics of organic transformations in a confined dendritic core: studies on the AIBN-initiated reaction of dendrimer cobalt(II) porphyrins with alkynes. AB - Cobalt(II) complexes of poly(aryl ester) dendrimer porphyrins [(m-[Gn]TPP)Co(II)] (generation number n=0-4), in the presence of azobisisobutyronitrile (AIBN) at 60 degrees C, underwent alkenylation with several alkynes at the metal center. A complete inhibition of double-bond migration (secondary transformation) was observed for [(m-[Gn]TPP)Co(II)] (n=3 and 4), which gave [(m-[Gn]TPP)Co(III) C(=CH(2))R] (n=3 and 4) exclusively. Overall reaction rates for [(m [Gn]TPP)Co(II)] (n=0-3) were hardly dependent on the size of the dendritic substituents, while a notable retardation was observed for the largest dendrimer, [(m-[G4]TPP)Co(II)]. Mechanistic studies on double-bond migration with pure [(m [Gn]TPP)Co(III)-C(=CH(2))Bu] (n=0-4) demonstrated that the secondary transformation involves participation of [(m-[Gn]TPP)Co(III)H] (n=0-4), derived from [(m-[Gn]TPP)Co(II)] and AIBN, rather than [(m-[Gn]TPP)Co(II)] alone. Crossover experiments using [(m-[Gn]TPP)Co(III)-C(=CH(2))Bu] (n=2-4), in combination with nondendritic [(m-[G0]TPP)Co(II)] and AIBN, indicated a high level of steric protection of the active center by a robust [G4]-dendritic cage, as suggested by a (1)H NMR pulse relaxation time profile of m-[G4]TPPH(2). PMID- 12898677 TI - The first successful crystallographic characterization of a cyclodextrin dimer: efficient synthesis and molecular geometry of a doubly sulfur-bridged beta cyclodextrin. AB - Beta-cyclodextrin is transannularly disulfonylated at the 6(A)- and 6(B) positions, and then converted to the corresponding 6(A),6(B)-diiodide and 6(A),6(B)-dithiol. Cross-coupling of the latter two species yields a single head to-head-coupled beta-cyclodextrin dimer 5 with two sulfur linkers at adjacent 6 methylene carbons. NMR and X-ray analysis revealed the trans-type ("aversive") linkage of both beta-cyclodextrin units. In the solid-state structure of 5.5 MeOH.23 H(2)O, the undistorted cyclodextrin macrocycles feature almost parallel ring planes pointing away from each other, leaving 5 with a "handcuff-like" appearance of approximate C(2) symmetry. This work represents the first successful crystallographic study on a cyclodextrin dimer. PMID- 12898678 TI - Covalent chemistry and conformational dynamics of topologically chiral amide based molecular knots. AB - The readily available in gram quantities tris(allyloxy)knot of the amide-type 5 (knotane) can be completely and partially deprotected with nBu(3)SnH in the presence of a palladium catalyst resulting in hydroxyknotanes 7-9. These, in turn, react with diethylchlorophosphate giving rise to knotanes equipped with between one and three phosphoryl groups. Sulfonylation of bis(allyloxy)monohydroxyknotane 8 with p-toluenesulfonyl chloride and, following removal of one or two allyl groups from the intermediate monosulfonate 13, give rise to sulfonyloxy-allyloxy-hydroxy- and sulfonyloxy-dihydroxy-knotanes 15 and 14, respectively. This provides a convenient method for the preparation of knotanes with any substitution pattern. All new knotanes have been isolated in preparative amounts and as highly pure substances with an exception of allyloxy dihydroxyknotane 9. This compound could only be obtained as a mixture with the corresponding monohydroxy-derivative 8. The structures of all synthesized compounds were established by means of FAB and MALDI TOF mass spectrometry, (1)H and (31)P NMR spectroscopy. The triphosphorylated knotane 10 exhibits high solubility in alcohols, allowing its complete enantiomeric resolution with a commercially available chiral HPLC column. (1)H,(1)H DQF-COSY correlation spectroscopy along with H/D exchange experiments and ab initio calculations provided the first detailed (1)H NMR signal assignments of knotanes in [D(6)]DMSO solution. The combination of variable temperature (1)H and (31)P NMR spectroscopy and molecular modeling has been applied to study the conformational behavior of the new knotanes in different solvents. It has been shown that in DMSO solution at room temperature knotanes exist in a relatively rigid nonsymmetrical conformation similar to that found in the solid state while faster conformational exchange leading to the average D(3) symmetrical structure was detected in a number of other solvents. PMID- 12898679 TI - Comparison of side-on and end-on coordination of E2 ligands in complexes [W(CO)5E2] (E=N, P, As, Sb, Bi, Si-, Ge-, Sn-, Pb-). AB - Complexes of W(CO)(5) with neutral diatomic pnictogen ligands N(2), P(2), As(2), Sb(2), and Bi(2) and anionic Group 14 ligands Si(2) (2-), Ge(2) (2-), Sn(2) (2-), and Pb(2) (2-) coordinated in both side-on and end-on fashion have been optimized by using density functional theory at the BP86 level with valence sets of TZP quality. The calculated bond energies have been used to compare the preferential binding modes of each respective ligand. The results were interpreted by analyzing the nature of the interaction between the ligands and the metal fragment using an energy partitioning method. This yields quantitative information regarding the strength of covalent and electrostatic interactions between the metal and ligand, as well as the contributions by orbitals of different symmetry to the covalent bonding. Results show that all the ligands studied bind preferentially in a side-on coordination mode, with the exception of N(2), which prefers to coordinate in an end-on mode. The preference of the heavier homologues P(2)-Bi(2) for binding in a side-on mode over the end-on mode in the neutral complexes [(CO)(5)WE(2)] comes mainly from the much stronger electrostatic attraction in the former species. The energy difference between the side-on and end-on isomers of the negatively charged complexes with the ligands Si(2) (2-), Ge(2) (2-), Sn(2) (2-), and Pb(2) (2-) is much less and it cannot be ascribed to a particular bonding component. PMID- 12898680 TI - Synthesis and properties of the first stable silylene-isocyanide complexes. AB - The first stable silylene-isocyanide complexes, [Tbt(Mes)SiCNAr] (5 c: Ar=Tip, 5 d: Ar=Tbt, 5 e: Ar=Mes*; Tbt=2,4,6-tris[bis(trimethylsilyl)methyl]phenyl, Mes=mesityl, Tip=2,4,6-triisopropylphenyl, Mes*=2,4,6-tri-tert-butylphenyl) were successfully synthesized by the reaction of a kinetically stabilized disilene, [Tbt(Mes)Si=Si(Mes)Tbt] (1), with bulky isocyanides, ArNC (3c-e). The spectroscopic data of 5 c-e and theoretical calculations for a model molecule indicated that 5 c-e are not classical cumulative compounds but the first stable silylene-Lewis base complexes. The reactions of 5 c-e with triethylsilane and 1,3 dienes gave the corresponding silylene adducts, and they underwent isocyanide exchange reactions in the presence of another isocyanide at room temperature. These results indicate dissociation of complexes 5 c-e to the corresponding silylene 2 and isocyanides 3 c-e under very mild conditions. The reaction of 5 c with methanol gave the MeOH adduct 16, [Tbt(Mes)SiHC(OMe)NTip], which has a hydrogen atom on the silicon atom. This regioselectivity can be explained in terms of the contribution of zwitterionic resonance structures D and E, which have an anion on the silicon atom. This result indicates that 5 c is not a classical cumulene having Si=C double bonds that should react with methanol to give adducts bearing a methoxyl group on the silicon atom. Although the reactions of 5 c-e with electrophilic reagents such as methanol, hydrogen chloride, and methyl iodide gave the formal silylene adducts, the studies on the reaction mechanism by trapping experiments and the observation of the intermediate suggested that the reaction mainly or partially proceeds by initial nucleophilic attack of the silicon atom, as is the case in the formation of 16 in the reaction of 5 c with methanol. It was revealed that 5 c-e show the nucleophilicity of the silicon atom, most likely resulting from the contribution of the zwitterionic resonance structures D and E. PMID- 12898681 TI - Electron delocalization in cross-conjugated p-phenylenevinylidene oligomers. AB - The synthesis, structure, and electronic properties of a series of cross conjugated p-phenylenevinylidene oligomers with one to four double bonds are reported. The X-ray crystal structure of the compound with two double bonds reveals a nonplanar conformation with torsion angles about the C(phenylene) C(vinylidene) and C(phenyl)-C(vinylidene) formal single bonds of 39.5(2) degrees and 30.5(2) degrees, respectively. Admixture of quinoid character in the ground state is observed. Infrared and Raman spectroscopy do not provide a clear picture of the degree of electron delocalization in the series, since the C=C stretching mode does not adequately reflect the C=C bond order and has a local nature. In contrast, electronic spectra and electrochemical data, as well as AM1 and PPP/SCF calculations, reveal that the cross-conjugated compounds basically behave as linearly pi-conjugated systems in the sense that molecular orbitals are delocalized over the entire structure and systematically change in energy. The electronic interaction between the repeating units is, however, not very strong, which has the consequence that spatial extension of the molecular orbitals does not lead to a red shift of the highest occupied molecular orbital-lowest unoccupied molecular orbital (HOMO-LUMO) electronic transition. This is related to the feature that the modest narrowing of the HOMO-LUMO gap with the chain length is accompanied by a relatively large reduction of electron repulsion. This finding implies that care should be taken in the use of electronic spectra for the evaluation of conjugation phenomena. PMID- 12898682 TI - GdIII complexes with fast water exchange and high thermodynamic stability: potential building blocks for high-relaxivity MRI contrast agents. AB - On the basis of structural considerations in the inner sphere of nine-coordinate, monohydrated Gd(III) poly(aminocarboxylate) complexes, we succeeded in accelerating the water exchange by inducing steric compression around the water binding site. We modified the common DTPA(5-) ligand (DTPA=(diethylenetriamine N,N,N',N",N"-pentaacetic acid) by replacing one (EPTPA(5-)) or two (DPTPA(5-)) ethylene bridges of the backbone by propylene bridges, or one coordinating acetate by a propionate arm (DTTA-prop(5-)). The ligand EPTPA(5-) was additionally functionalized with a nitrobenzyl linker group (EPTPA-bz-NO(2) (5-)) to allow for coupling of the chelate to macromolecules. The water exchange rate, determined from a combined variable-temperature (17)O NMR and EPR study, is two orders of magnitude higher on [Gd(eptpa-bz-NO(2))(H(2)O)](2-) and [Gd(eptpa)(H(2)O)](2-) than on [Gd(dtpa)(H(2)O)](2-) (k(ex)298=150x10(6), 330x10(6), and 3.3x10(6) s(-1), respectively). This is optimal for attaining maximum proton relaxivities for Gd(III)-based, macrocyclic MRI contrast agents. The activation volume of the water exchange, measured by variable-pressure (17)O NMR spectroscopy, evidences a dissociative interchange mechanism for [Gd(eptpa)(H(2)O)](2-) (DeltaV(not equal sign)=(+6.6+/-1.0) cm(3) mol(-1)). In contrast to [Gd(eptpa)(H(2)O)](2-), an interchange mechanism is proved for the macrocyclic [Gd(trita)(H(2)O)](-) (DeltaV (not equal sign)=(-1.5+/-1.0) cm(3) mol(-1)), which has one more CH(2) group in the macrocycle than the commercial MRI contrast agent [Gd(dota)(H(2)O)](-), and for which the elongation of the amine backbone also resulted in a remarkably fast water exchange. When one acetate of DTPA(5-) is substituted by a propionate, the water exchange rate on the Gd(III) complex increases by a factor of 10 (k(ex)298=31x10(6) s(-1)). The [Gd(dptpa)](2-) chelate has no inner-sphere water molecule. The protonation constants of the EPTPA-bz-NO(2) (5-) and DPTPA(5-) ligands and the stability constants of their complexes with Gd(III), Zn(II), Cu(II) and Ca(II) were determined by pH potentiometry. Although the thermodynamic stability of [Gd(eptpa bz-NO(2))(H(2)O)](2-) is reduced to a slight extent in comparison with [Gd(dtpa)(H(2)O)](2-), it is stable enough to be used in medical diagnostics as an MRI contrast agent. Therefore both this chelate and [Gd(trita)(H(2)O)](-) are potential building blocks for the development of high-relaxivity macromolecular agents. PMID- 12898683 TI - Lipopeptaibol metabolites of tolypocladium geodes: total synthesis, preferred conformation, and membrane activity. AB - We have synthesized by solution methods and characterized the lipopeptaibol metabolite LP237-F8 extracted from the fungus Tolypocladium geodes and five selected analogues with the Etn-->Aib or Etn-->Nva replacement at position 8 and/or a triple Gln-->Glu(OMe) replacement at positions 5, 6, and 9 (Etn=Calpha ethylnorvaline, Aib=alpha-aminoisobutyric acid, Nva=norvaline). Conformation analysis, performed by FT-IR absorption, NMR, and CD techniques, strongly supports the view that the six terminally blocked decapeptides are highly helical in solution. Helix topology and amphiphilic character are responsible for their remarkable membrane activity. At position 8 the combination of high hydrophobicity and Calpha tetrasubstitution, as in the Etn-containing LP237-F8 metabolite, has a positive effect on membrane interaction. PMID- 12898684 TI - N-Heterocyclic carbene functionalized iridium phosphinidene complex [Cp*(NHC)Ir=PMes*]: comparison of phosphinidene, imido, and carbene complexes. AB - The novel phosphinidene complex [Cp*(NHC)Ir=PMes*] (3; NHC=1,3-diisopropyl-4,5 dimethylimidazol-2-ylidene) was prepared in high yield from [Cp*(NHC)IrCl(2)] (2) and [LiPHMes*].3 THF. It represents the first example of an NHC ligated transition metal phosphinidene complex. The X-ray crystal structure for 3 is also reported. DFT calculations on the N-heterocyclic carbene containing parent complexes [Cp(NHC)Ir=E] (E=PH, NH, CH(2)) show that the NHC ligand acts as good sigma-donor/weak pi-acceptor ligand and forms strong Ir-C(NHC) single bonds. The Ir=E double bonds result from strong triplet-triplet interactions between [Cp(NHC)Ir] and E. PMID- 12898685 TI - Tuning the redox potentials of dinuclear tungsten oxo complexes [(Cp*W(4,4'-R,R 2,2'-bpy)(mu-O))2][PF6]2 toward photochemical water splitting. AB - A series of novel dinuclear tungsten(IV) oxo complexes with disubstituted 4,4' R,R-2,2'-bipyridyl (R(2)bpy) ligands of the type [(Cp*W(R(2)bpy)(mu O))(2)][PF(6)](2) (R=NMe(2), tBu, Me, H, Cl) was prepared by hydrolysis of the tungsten(IV) trichloro complexes [Cp*W(R(2)bpy)Cl(3)]. Cyclic voltammetry measurements for the tungsten(IV) oxo compounds provided evidence for one reversible oxidation and two reversible reductions leading to the oxidation states W(V)W(IV), W(IV)W(III) and W(III)W(III). The corresponding complexes [(Cp*W(R(2)bpy)(mu-O))(2)](n+) [PF(6)](n) (n=0 for R=Me, tBu, and 1, 3 for both R=Me) could be isolated after chemical oxidation/reduction of the tungsten(IV) oxo complexes. The crystal structures of the complexes [(Cp*W(R(2)bpy)(mu O))(2)][BPh(4)](2) (R=NMe(2), tBu) and [(Cp*W(Me(2)bpy)(mu-O))(2)](n+)[PF(6)](n) (n=0, 1, 2, 3) show a cis geometry with a puckered W(2)O(2) four-membered ring for all compounds except [(Cp*W(Me(2)bpy)(mu-O))(2)] which displays a trans geometry with a planar W(2)O(2) ring. Examining the interaction of these novel tungsten oxo complexes with protons, we were able to show that the W(IV)W(IV) complexes [(Cp*W(R(2)bpy)(mu-O))(2)][PF(6) (-)](2) (R=NMe(2), tBu) undergo reversible protonation, while the W(III)W(III) complexes [(Cp*W(R(2)bpy)(mu O))(2)] transfer two electrons forming the W(IV)W(IV) complex and molecular hydrogen. PMID- 12898686 TI - Stabilization of CoI by ZnII in pure acetonitrile and its reaction with aryl halides. AB - The study of the electrochemical behavior of cobalt(II) bromide (CoBr(2)) in pure acetonitrile allowed us to demonstrate that Co(2+) is the catalyst precursor involved in the electrochemical and chemical conversions of arylhalides, ArX, to arylzinc compounds in that solvent. The reduction of Co(2+) leads to the Co(+) species, which disproportionates too rapidly to react further with aryl halides. However, the presence of zinc(II) bromide allows us to stabilize the electrogenerated cobalt(I) and to observe it on the timescale of slow cyclic voltammetry. Under such conditions, the Co(I) species has time to react with aryl halides and produce [Co(III)ArX](+) complexes that are reduced into [Co(II)ArX] by a single electron uptake at the same potential at which Co(2+) is reduced. Rate constants for the oxidative addition of ArX to Co(I) have been determined for various aryl halides and compared to the values obtained in an acetonitrile (ACN)/pyridine (9:1, v/v) mixture. It is shown that Co(I) is stabilized more by ZnBr(2) than by pyridine. A transmetallation reaction between [Co(II)ArX] and ZnBr(2) has also been observed. We finally propose a mechanism for the cobalt catalyzed electrochemical conversion of aryl bromides into organozinc species in pure acetonitrile. PMID- 12898687 TI - Cofactor recycling mechanism in asymmetric biocatalytic reduction of carbonyl compounds mediated by yeast: which is the efficient electron donor? AB - In asymmetric reduction of carbonyl compounds mediated by microorganisms, the cofactors that transfer hydride should be regenerated by using a recycling system. In most cases, this recycling system consists of carbohydrate molecules, especially glucose or sucrose. Other molecules such as ethanol and acetate have been used as electron donors too. The reduction can even be conducted without added electron donors. To improve biocatalytic synthesis, it is important to understand the cofactor recycling mechanism. In this work, the hydride-transfer mechanism in cofactor regeneration, which takes place in bioreduction mediated by yeast, was studied by means of an isotope tracing technique. The results show that, when glucose was used, the NADH involved in the glycolysis was consumed directly in the formation of ethanol and was not used in the bioreduction. Hence, the regeneration of cofactors in the reduction is not coupled with glycolysis. Nevertheless, glucose is an efficient electron donor that transfers hydride through the hexose monophosphate (HMP) pathway in which the main hydrogen source is C-1 and C-3 hydrogen of glucose. Ethanol is not a good electron donor, since, when it was used, only a small quantity of hydrogen was transferred from this molecule, and the main hydrogen source was water. Therefore, the ethanol oxidation pathway may not be efficient. In the absence of added auxiliary substrates, the yeast cells may use electron donors stored in its cellules. However, in this case we observed that the main hydrogen source for cofactor recycling was water, while only very few hydrogen atoms were from unexchangeable sites. This is similar to the case in which ethanol is used, and is in contradiction with the HMP pathway if stored glucose was the electron donor. The question that remains to be investigated is "what is the efficient electron donor recycling mechanism in the yeast cellules?" PMID- 12898688 TI - On the electronic structures of the 1,3-diboracyclobutane-1,3-diyls and their valence isomers with a B2E2 skeleton (E=N, P, As). AB - The concept of through-space versus through-bond interactions on the stabilization of biradical structures with a singlet or triplet ground state is evaluated for the 1,3-diboracyclobutane-1,3-diyls and related congeners. Singlet biradicals are favored when the intermediate units E feature singlet character (PH(2) (+), AsH(2) (+)), while E fragments with triplet character (NH(2) (+)) induce small energy separations between the lowest singlet and triplet states. These considerations are supported by quantum chemical calculations with energy optimization at 1) MCSCF level plus MR-MP2 correction, 2) MR-MP2 level, and 3) two different types of density functional levels for the planar (D(2h)) geometries. The singlet-triplet energy separations in the planar compounds increase with increasing singlet stability of the corresponding E fragments. In addition to this newly developed principal features for singlet stabilization, which primarily occurs in bonded structures with higher main-group elements, the corresponding valence isomers with bicyclobutane, cyclobutene and cis-butadiene structures are investigated. PMID- 12898689 TI - Formation of dinuclear titanium and zirconium complexes by olefin metathesis- catalytic preparation of organometallic catalyst systems. AB - The titanium complex [(C(5)H(4)bond;allyl)TiCl(3)] (2) undergoes olefin metathesis coupling when treated with 3 mol % of [Cl(2)(L(1))(L(2))Ru=CHPh] (L(1)=L(2)=PCy(3), 4 a; L(1)=PCy(3), L(2)=(H(2)IMes), 4 b) to yield the dimetallic complex [Cl(3)Ti(C(5)H(4))-CH(2)CH=CHCH(2)-(C(5)H(4))TiCl(3)] (5). The allyl-substituted titanocene complex [Cp(C(5)H(4)bond;allyl)TiCl(2)] (3) analogously yields the dimetallic system 6 when treated with 4. The ansa zirconocene complex [Me(2)Si(C(5)H(4))(C(5)H(3)bond;allyl)ZrCl(2)] (7) cleanly yields the analogous dimetallic coupling product 8 (>95 % isomerically pure), when treated with catalytic amounts of 4 b in toluene. Complex 8 gives an active homogeneous ethene or propene polymerization catalyst, especially at elevated temperatures, when treated with excess methylalumoxane. PMID- 12898690 TI - High-pressure preparation, crystal structure, and properties of alpha-(RE)2B4O9 (RE=Eu, Gd, Tb, Dy): oxoborates displaying a new type of structure with edge sharing BO4 tetrahedra. AB - High-pressure/high-temperature conditions of 10 GPa and 1150 degrees C were used to synthesize the new rare-earth oxoborates alpha-(RE)(2)B(4)O(9) (RE=Eu, Gd, Tb, Dy) in a Walker-type multianvil apparatus. Single-crystal X-ray structure determination of alpha-(RE)(2)B(4)O(9) (RE=Eu, Gd, Tb) revealed: C2/c, Z=20, alpha-Eu(2)B(4)O(9): a=2547.9(5), b=444.3(1), c=2493.8(5) pm, beta=99.82(3) degrees, R1=0.0277, wR2=0.0693 (all data); alpha-Gd(2)B(4)O(9): a=2539.0(1), b=443.3(1), c=2490.8(1) pm, beta=99.88(1) degrees, R1=0.0457, wR2=0.0643 (all data); alpha-Tb(2)B(4)O(9): a=2529.4(1), b=441.6(1), c=2484.3(1) pm, beta=99.88(1) degrees, R1=0.0474, wR2=0.0543 (all data). The isotypic compounds exhibit a new type of structure that is built up of BO(4) tetrahedra to form a network that incorporates the rare-earth cations. The most important feature is the existence of the new structural motif of edge-sharing BO(4) tetrahedra next to the known motif of corner-sharing BO(4) tetrahedra, which is realized in the presented compounds alpha-(RE)(2)B(4)O(9) (RE=Eu, Gd, Tb, Dy) for the second time. Furthermore, we report the temperature-resolved in-situ powder-diffraction measurements, DTA, IR/Raman spectroscopic investigations, and magnetic properties of the new compounds. PMID- 12898691 TI - Synthesis and reactivity of calix[4]arene-supported group 4 imido complexes. AB - New mononuclear titanium and zirconium imido complexes [M(NR)(R'(2)calix)] [M=Ti, R'=Me, R=tBu (1), R=2,6-C(6)H(3)Me(2) (2), R=2,6-C(6)H(3)iPr(2) (3), R=2,4,6 C(6)H(2)Me(3) (4); M=Ti, R'=Bz, R=tBu (5), R=2,6-C(6)H(3)Me(2) (6), R=2,6 C(6)H(3)iPr(2) (7); M=Zr, R'=Me, R=2,6-C(6)H(3)iPr(2) (8)] supported by 1,3 diorganyl ether p-tert-butylcalix[4]arenes (R'(2)calix) were prepared in good yield from the readily available complexes [MCl(2)(Me(2)calix)], [Ti(NR)Cl(2)(py)(3)], and [Ti(NR)Cl(2)(NHMe(2))(2)]. The crystallographically characterised complex [Ti(NtBu)(Me(2)calix)] (1) reacts readily with CO(2), CS(2), and p-tolyl-isocyanate to give the isolated complexes [Ti[N(tBu)C(O)O](Me(2)calix)] (10), [[Ti(mu-O)(Me(2)calix)](2)] (11), [[Ti(mu S)(Me(2)calix)](2)] (12), and [Ti[N(tBu)C(O)N(-4-C(6)H(4)Me)](Me(2)calix)] (13). In the case of CO(2) and CS(2), the addition of the heterocumulene to the Ti-N multiple bond is followed by a cycloreversion reaction to give the dinuclear complexes 11 and 12. The X-ray structure of 13.4(C(7)H(8)) clearly establishes the N,N'-coordination mode of the ureate ligand in this compound. Complex 1 undergoes tert-butyl/arylamine exchange reactions to form 2, 3, [Ti(N-4 C(6)H(4)Me)(Me(2)calix)] (14), [Ti(N-4-C(6)H(4)Fc)(Me(2)calix)] (15) [Fc=Fe(eta(5)-C(5)H(5))(eta(5)-C(5)H(4))], and [[Ti(Me(2)calix)](2)[mu-(N-4 C(6)H(4))(2)CH(2)]] (16). Reaction of 1 with H(2)O, H(2)S and HCl afforded the compounds [[Ti(mu-O)(Me(2)calix)](2)] (11), [[Ti(mu-S)(Me(2)calix)](2)] (12), and [TiCl(2)(Me(2)calix)] in excellent yields. Furthermore, treatment of 1 with two equivalents of phenols results in the formation of [Ti(O-4 C(6)H(4)R)(2)(Me(2)calix)] (R=Me 17 or tBu 18), [Ti(O-2,6 C(6)H(3)Me(2))(2)(Me(2)calix)] (19) and [Ti(mbmp)(Me(2)calix)] (20; H(2)mbmp=2,2' methylene-bis(4-methyl-6-tert-butylphenol) or CH(2)([CH(3)][C(4)H(9)]C(6)H(2) OH)(2)). The bis(phenolate) compounds 17 and 18 with para-substituted phenolate ligands undergo elimination and/or rearrangement reactions in the nonpolar solvents pentane or hexane. The metal-containing products of the elimination reactions are dinuclear complexes [[Ti(O-4-C(6)H(4)R)(Mecalix)](2)] [R=Me (23) or tBu (24)] where Mecalix=monomethyl ether of p-tert-butylcalix[4]arene. The products of the rearrangement reaction are [Ti(O-4-C(6)H(4)Me)(2) (paco Me(2)calix)] (25) and [Ti(O-4-C(6)H(4)tBu)(2)(paco-Me(2)calix)] (26), in which the metallated calix[4]arene ligand is coordinated in a form reminiscent of the partial cone (paco) conformation of calix[4]arene. In these compounds, one of the methoxy groups is located inside the cavity of the calix[4]arene ligand. The complexes 24, 25 and 26 have been crystallographically characterised. Complexes with sterically more demanding phenolate ligands, namely 19 and 20 and the analogous zirconium complexes [Zr(O-4-C(6)H(4)Me)(2)(Me(2)calix)] (21) and [Zr(O 2,6-C(6)H(3)Me(2))(2)(Me(2)calix)] (22) do not rearrange. Density functional calculations for the model complexes [M(OC(6)H(5))(2)(Me(2)calix)] with the calixarene possessing either cone or partial cone conformations are briefly presented. PMID- 12898692 TI - Synthesis of a new potential tridentate anthracene ligand bearing deprotectable methoxymethoxy (OMOM) group at 1,8-positions: attempt to synthesize anionic hypervalent carbon compounds. AB - A novel potential tridentate ligand bearing deprotectable coordinating atoms, 1,8 bis(methoxymethoxy)-9-bromoanthracene (15), was synthesized. The key steps are as follows: 1) stepwise mono-oxygenation from 1,8-dibromo-9-methoxyanthracene by use of electrophilic oxaziridine and gaseous dioxygen, and 2) selective reduction of the methoxy group by LDBB (lithium di-tert-butylbiphenylide) followed by treatment with BrCF(2)CF(2)Br. The corresponding 1,8-bis(methoxymethoxy)-9-lithio anthracene (14), which should be a useful versatile trianion equivalent, could be generated by treatment of the bromide with one equivalent of nBuLi. The lithioanthracene reacted with hexafluoroacetone to give the deprotected ether 17 together with the adduct alcohol 16. The ether could easily be deprotected to give 1,2-dihydro-1,1-bis(trifluoromethyl)-2-oxa-9-hydroxyanthrylene (8), which was deprotonated with KH in the presence of [18]crown-6 to give the corresponding anion (9-K([18]crown-6)). The X-ray structure and NMR spectra of 9-K([18]crown-6) showed that it has an unsymmetrical structure probably due to the interaction between the oxygen atom of the phenoxide and the potassium cation surrounded by the crown ether. PMID- 12898693 TI - Construction of artificial signal transducers on a lectin surface by post photoaffinity-labeling modification for fluorescent saccharide biosensors. AB - A new general method, post-photoaffinity-labeling modification (PPALM), for constructing fluorescent saccharide biosensors based on naturally occurring saccharide-binding proteins, lectins, is described in detail. An active-site directed incorporation of a masked reactive site into a lectin was conducted by using a photoaffinity labeling technique followed by demasking and then chemical modification to yield a fluorescent lectin. Two photoaffinity labeling reagents were designed and synthesized in this study. The labeling reagent with a photoreactive site appended through a disulfide link to a mannoside unit was bound to the saccharide-binding pocket of the lectin concanavalin A (Con A). After light irradiation, the mannoside unit was cleaved by reduction. The unique thiol group thus produced was site-specifically modified with various fluorescent groups (dansyl, coumarin, or dimethylaminobenzoate derivatives) to afford fluorescent Con As. The labeling site was characterized by protease-catalyzed digestion followed by HPLC, MALDI-TOF MS, and tandem mass-mass spectrometry; these methods indicated that the photolabeling step is remarkably site specific. Strong fluorescence was observed in the engineered Con A with a fluorophore, and the emission changed sensitively upon saccharide complexation. The binding constants for various saccharides were determined by fluorescence titration and demonstrated that the binding selectivity and affinity of the engineered Con As are comparable to those of native Con A. The red shift of the emission maximum, the decrease in the fluorescence anisotropy of the dansyl unit, and the increase in the twisted intramolecular charge transfer emission caused by sugar binding to the engineered Con A explicitly indicate that the microenvironment of the appended fluorophores changes from a restricted and relatively hydrophobic environment into a rather freely mobile and hydrophilic environment. PMID- 12898694 TI - Vibrational and quantum-chemical study of push-pull chromophores for second-order nonlinear optics from rigidified thiophene-based pi-conjugating spacers. AB - Two types of push-pull chromophores built around thiophene-based pi-conjugating spacers rigidified by either covalent bonds or noncovalent intramolecular interactions have been analysed by means of IR and Raman spectroscopical measurements in the solid state as well as in a variety of solvents. Comparison of the Raman features of NLO-phores based on a covalently rigidified dithienylene (DTE) spacer with those of their open chain DTE analogues shows that the bridging of the central double bond of DTE with the nearest beta-positions of the thienyl units through two ethylene bridges significantly improves the intramolecular charge transfer. This also occurs for NLO-phores based on a 2,2'-bi(3,4 ethylenedioxythiophene) (BEDOT) spacer as compared with their corresponding parent compounds based on an unsubstituted bithiophene (BT) spacer. For NLO phores based on a BEDOT spacer, noncovalent intramolecular interactions between sulfur and oxygen atoms are responsible for the rigidification of the spacer. The Raman spectra of these NLO-phores obtained in the form of solutes in dilute solutions reveal two different behaviours: i) chromophores based on covalently bridged or open chain DTE spacers display Raman spectral profiles in solution quite similar to those of the corresponding solids, with a very little dependence on the polarity of the solvent, while ii) larger spectral changes are noticed for NLO-phores built around BEDOT or BT spacers on going from solids to solutions. In the second case, spectral changes must be ascribed not solely to conformational distortions of the donor and acceptor end groups with respect to the pi conjugated backbone mean-square-plane (as for the DTE-based NLO-phores) but also to distortions of the thienyl units of the pi-conjugating spacer from coplanarity. The insertion of vinylenic bridges between the thienyl units of the pi-conjugating spacer and between the spacer and the donor and acceptor end groups is a suitable strategy to reach a fairly large intramolecular charge transfer both in polar and nonpolar solvents. Density functional theory (DFT) calculations have been carried out to assign the relevant electronic and vibrational features and to derive useful information about the molecular structure of these NLO-phores. PMID- 12898695 TI - The synthesis of acid- and base-labile lipopeptides on solid support. AB - Lipidated peptides, including characteristic partial structures of human Ras proteins, were synthesized by means of a new solid-phase technique in 22-68 % yield. This technique gives access to farnesylated, palmitoylated, and doubly lipidated peptides as methyl esters or carboxylic acids carrying a fluorescent tag or a maleimide moiety for coupling to proteins. The peptide backbones were built up on the resin by using 9-fluorenylmethoxycarbonyl chemistry together with the oxidatively cleavable hydrazide linker. As a key step, the acid-labile farnesyl and basic-labile palmitoyl lipid groups were introduced onto the resin after the cleavage of appropriate acid- or reduction-sensitive protecting groups from the cysteine residues. Optional introduction of different fluorescent tags or a maleimide group into the peptide was followed by release of the resin-bound target peptide as the methyl ester or carboxylic acid by very mild copper(II) mediated oxidation in slightly acidic or basic media. This new methodology should substantially facilitate the access to lipidated peptides for the study of important biological phenomena like biological signal transduction, localization, and vesicular transport. PMID- 12898696 TI - The role of cystine knots in collagen folding and stability, part I. Conformational properties of (Pro-Hyp-Gly)5 and (Pro-(4S)-FPro-Gly)5 model trimers with an artificial cystine knot. AB - In analogy to the cystine knots present in natural collagens, a simplified disulfide cross-link was used to analyse the conformational effects of a C terminal artificial cystine knot on the folding of collagenous peptides consisting of solely (Pro-Hyp-Gly) repeating units. Assembly of the alpha chains into a heterotrimer by previously applied regioselective disulfide-bridging strategies failed because of the high tendency of (Pro-Hyp-Gly)(5) peptides to self-associate and form homotrimers. Only when side-chain-protected peptides were used, for example in the Hyp(tBu) form, and a new protection scheme was adopted, selective interchain-disulfide cross-linking into the heterotrimer in organic solvents was successful. This unexpected strong effect of the conformational properties on the efficiency of well-established reactions was further supported by replacing the Hyp residues with (4S)-fluoroproline, which is known to destabilise triple-helical structures. With the related [Pro-(4S)-FPro-Gly](5) peptides, assembly of the heterotrimer in aqueous solution proceeded in a satisfactory manner. Both the intermediates and the final fluorinated heterotrimer are fully unfolded in aqueous solution even at 4 degrees C. Conversely, the disulfide-crossbridged (Pro-Hyp-Gly)(5) heterotrimer forms a very stable triple helix. The observation that thermal unfolding leads to scrambling of the disulfide bridges was unexpected. Although NMR experiments support an extension of the triple helix into the cystine knot, thermolysis is not associated with the unfolding process. In fact, the unstructured fluorinated trimer undergoes an equally facile thermodegradation associated with the intrinsic tendency of unsymmetrical disulfides to disproportionate into symmetrical disulfides under favourable conditions. The experimental results obtained with the model peptides fully support the role of triple-helix nucleation and stabilisation by the artificial cystine knot as previously suggested for the natural cystine knots in collagens. PMID- 12898697 TI - The role of cystine knots in collagen folding and stability, part II. Conformational properties of (Pro-Hyp-Gly)n model trimers with N- and C-terminal collagen type III cystine knots. AB - In mature collagen type III the homotrimer is C-terminally cross-linked by an interchain cystine knot consisting of three disulfide bridges of unknown connectivity. This cystine knot with two adjacent cysteine residues on each of the three alpha chains has recently been used for the synthesis and expression of model homotrimers. To investigate the origin of correct interchain cysteine pairings, (Pro-Hyp-Gly)(n) peptides of increasing triplet number and containing the biscysteinyl sequence C- and N-terminally were synthesised. The possibilities were that this origin may be thermodynamically coupled to the formation of the collagen triple helix as happens in the oxidative folding of proteins, or it could represent a post-folding event. Only with five triplets, which is known to represent the minimum number for self-association of collagenous peptides into a triple helix, air-oxidation produces the homotrimer in good yields (70 %), the rest being intrachain oxidised monomers. Increasing the number of triplets has no effect on yield suggesting the formation of kinetically trapped intermediates, which are not reshuffled by the glutathione redox buffer. N-terminal incorporation of the cystine knot is significantly less efficient in the homotrimerisation step and also in terms of triple-helix stabilisation. Compared to an artificial C-terminal cystine knot consisting of two interchain disulfide bridges, the collagen type III cystine knot produces collagenous homotrimers of remarkably high thermostability, although the concentration-independent refolding rates are not affected by the type of disulfide bridging. Since the natural cystine knot allows ready access to homotrimeric collagenous peptides of significantly enhanced triple-helix thermostability it may well represent a promising approach for the preparation of collagen-like innovative biomaterials. Conversely, the more laborious regioselectively formed artificial cystine knot still represents the only synthetic strategy for heterotrimeric collagenous peptides. PMID- 12898699 TI - Evidence for astrocyte heterogeneity: a distinct subpopulation of protoplasmic like glial cells is detected in transgenic mice expressing Lmo1-lacZ. AB - The adult mammalian central nervous system (CNS) contains a large number of different cell types, which arise from the ventricular (VZ) and subventricular zones during embryonic development. In this study, we used a transgenic mouse expressing Lmo1-LacZ from a randomly inserted promoter/reporter gene construct to identify a glial subpopulation. LMO1 is an LIM domain-containing protein, thought to act in protein-protein interactions. We found first that in the adult transgenic CNS, beta-galactosidase (beta-gal) was expressed in a specific subpopulation of protoplasmic-like cells, which did not express detectable levels of glial fibrilary acidic protein unless a lesion was produced. Secondly, during development, beta-gal(+) cells were found arising from discrete regions of the VZ. Taken together, these results identify a subpopulation of protoplasmic glial cells in the adult CNS and suggest that they arise from a restricted VZ region during CNS development. PMID- 12898700 TI - Expression and regulation of L-cystine transporter, system xc-, in the newly developed rat retinal Muller cell line (TR-MUL). AB - The purpose of the present study was to elucidate the expression and regulation of the L-cystine transporter, system x(c) (-), in Muller cells. In this study, newly developed conditionally immortalized rat Muller cell lines (TR-MUL) from transgenic rats harboring the temperature-sensitive SV 40 large T-antigen gene were used as an in vitro model. TR-MUL cells express large T-antigen and grow well at 33 degrees C with a doubling time of 30 h, but do not grow at 39 degrees C. TR-MUL cells express typical Muller cell markers such as S-100, glutamine synthetase, and EAAT1/GLAST, whereas EAAT2/GLT-1 and EAAT5 are not detected. TR MUL cells also exhibit little or no expression of glial fibrillary acidic protein. We found that TR-MUL5 cells exhibited [(14)C]L-cystine uptake activity and expressed xCT and 4F2hc, which involve system x(c) (-). The uptake of [(14)C]L-cystine was significantly inhibited by L-glutamic acid and L-aspartic acid, whereas L-leucine had no effect. Following diethyl maleate (DEM) treatment, the glutathione concentration in TR-MUL5 cells was reduced in the first 24 h, then gradually recovered for more than 24 h. The L-cystine uptake rate and the xCT expression level in TR-MUL5 cells were enhanced by DEM treatment. In contrast, the 4F2hc expression level was unchanged. In conclusion, TR-MUL cells have the properties of Muller cells and exhibit system x(c) (-)-mediated L cystine uptake activity. The oxidative stress conditions following DEM treatment activate L-cystine transport in TR-MUL cells due to the enhanced transcription of the xCT gene. PMID- 12898701 TI - Nucleotide-mediated calcium signaling in rat cortical astrocytes: Role of P2X and P2Y receptors. AB - ATP is the dominant messenger for astrocyte-to-astrocyte calcium-mediated communication. Definition of the exact ATP/P2 receptors in astrocytes and of their coupling to intracellular calcium ([Ca(2+)](i)) has important implications for brain physiology and pathology. We show that, with the only exception of the P2X(6) receptor, primary rat cortical astrocytes express all cloned ligand-gated P2X (i.e., P2X(1-5) and P2X(7)) and G-protein-coupled P2Y receptors (i.e., P2Y(1), P2Y(2), P2Y(4), P2Y(6), and P2Y(12)). These cells also express the P2Y like UDP-glucose receptor, which has been recently recognized as the P2Y(14) receptor. Single-cell image analysis showed that only some of these receptors are coupled to [Ca(2+)](i). While ATP induced rapid and transient [Ca(2+)](i) increases (counteracted by the P2 antagonists suramin, pyridoxal-phosphate-6 azophenyl-2'-4'-disulfonic acid and oxidized ATP), the P2X(1)/P2X(3) agonist alphabetameATP produced no changes. Conversely, the P2X(7) agonist BzATP markedly increased [Ca(2+)](i); the presence and function of the P2X(7) receptor was also confirmed by the formation of the P2X(7) pore. ADP and 2meSADP also produced [Ca(2+)](i) increases antagonized by the P2Y(1) antagonist MRS2179. Some cells also responded to UTP but not to UDP. Significant responses to sugar-nucleotides were also detected, which represents the first functional response reported for the putative P2Y(14) receptor in a native system. Based on agonist preference of known P2 receptors, we conclude that, in rat astrocytes, ATP-induced calcium rises are at least mediated by P2X(7) and P2Y(1) receptors; additional receptors (i.e., P2X(2), P2X(4), P2X(5), P2Y(2), P2Y(4), and P2Y(14)) may also contribute. PMID- 12898702 TI - Phagocytosis of apoptotic inflammatory cells by microglia and its therapeutic implications: termination of CNS autoimmune inflammation and modulation by interferon-beta. AB - Apoptosis of autoaggressive T-cells in the CNS is an effective, noninflammatory mechanism for the resolution of T-cell infiltrates, contributing to clinical recovery in T-cell-mediated neuroinflammatory diseases. The clearance of apoptotic leukocytes by tissue-specific phagocytes is critical in the resolution of the inflammatory infiltrate and leads to a profound downregulation of phagocyte immune functions. Adult human microglia from surgically removed normal brain tissue was used in a standardized, light-microscopic in vitro phagocytosis assay of apoptotic autologous peripheral blood-derived mononuclear cells (MNCs). Microglia from five different patients had a high capacity for the uptake of apoptotic MNCs in contrast to nonapoptotic target cells with the phagocytosis rate for nonapoptotic MNCs amounting to only 61.6% of the apoptotic MNCs. A newly described phosphatidylserine receptor, critical in the phagocytosis of apoptotic cells by macrophages, is also expressed at similar levels on human microglia. The effects of the therapeutically used immunomodulatory agent interferon-beta (IFNbeta) were investigated using Lewis rat microglia and apoptotic, encephalitogenic, myelin basic protein-specific autologous T-cells. Also, rat microglia had a high capacity to phagocytose apoptotic T-cells specifically. IFNbeta increased the phagocytosis of apoptotic T-cells to 36.8% above the untreated controls. The enhanced phagocytic activity was selective for apoptotic T-cells and was not mediated by increased IL-10 secretion. Apoptotic inflammatory cells may be efficiently and rapidly removed by microglial cells in the autoimmune-inflamed human CNS. The in vitro increase of phagocytosis by IFNbeta merits further investigations whether this mechanism could also be therapeutically exploited. PMID- 12898703 TI - Cytokine, chemokine and growth factor gene profiling of cultured human astrocytes after exposure to proinflammatory stimuli. AB - Astrocytes play key roles in CNS development, inflammation, and repair by producing a wide variety of cytokines, chemokines, and growth factors. Understanding the regulation of this network is important for a full understanding of astrocyte functioning. In this study, expression levels of 268 genes encoding cytokines, chemokines, growth factors, and their receptors were established in cultured human adult astrocytes using cDNA arrays. Also, changes in this gene profile were determined following stimulation with TNFalpha, IL 1beta, and IFNgamma. The data obtained reveal a highly reproducible pattern of gene expression not only between different astrocyte cultures from a single source, but also between astrocytes from different donors. They also identify several gene products not previously described for human astrocytes, including a.o. IL-17, CD70, CD147, and BIGH3. When stimulated with TNFalpha astrocytes respond with increased expression of several genes, notably including those encoding the chemokines CCL2 (MCP-1), CCL5 (RANTES), and CXCL8 (IL-8), growth factors including BMP-2A, BMP-3, neuromodulin (GAP43), BDNF, and G-CSF, and receptors such as the CRF receptor, the calcitonin receptor (CTR), and TKT. The response to IL-1beta involves largely the same range of genes, but responses were blunted in comparison to the TNFalpha response. Treatment with IFNgamma had no or only marginal effects on expression of any of the 268 genes analyzed. Astrocytes treated with a mixture of all three stimuli together displayed responses that are largely similar to those found in response to TNFalpha or IL-1beta alone, with only few additional synergistic effects. PMID- 12898704 TI - Essential role for ERK mitogen-activated protein kinase in matrix metalloproteinase-9 regulation in rat cortical astrocytes. AB - Matrix metalloproteinases (MMPs) contribute to the pathophysiology of brain injury and inflammation but little is known about their regulatory signaling pathways in brain cells. Here we examine the role of mitogen-activated protein (MAP) kinase pathways in MMP-9 regulation in cortical rat astrocytes. The protein kinase C (PKC) activator phorbol 12-myristate 13-acetate (PMA) induced MMP-9 but not MMP-2 secretion as measured by gelatin zymography. Northern blot and RT-PCR analysis showed that MMP-9 responses occurred at the mRNA level. Although PMA increased phosphorylation in all three major MAP kinase pathways (ERK, p38 MAP kinase, and JNK), only inhibition of the ERK pathway by the MEK/ERK inhibitor U0126 (0.1-10 microM) significantly reduced MMP-9 upregulation, even when treatment was delayed for 4 h after PMA exposure. Inhibitors of p38 MAP kinase (SB203580) and JNK (SP600125) had no effect. This PKC pathway was compared to a cytokine response by exposing astrocytes to TNFalpha, which also activated MAP kinase and induced MMP-9 upregulation. But in this case, all three MAP kinase inhibitors (U0126, SB203580, and SP600125) reduced TNFalpha-induced MMP-9 upregulation. Taken together, these results suggest that the ERK MAP kinase is essential for MMP-9 upregulation via PKC and cytokine pathways in astrocytes. PMID- 12898705 TI - Modulation of intercellular calcium signaling in astrocytes by extracellular calcium and magnesium. AB - The extracellular concentrations of Ca(2+) and Mg(2+) are well known to play important roles in the function of the central nervous system. We examined the effects of extracellular Ca(2+) and Mg(2+) on ATP release and intercellular signaling in astrocytes. The extent of propagation of intercellular Ca(2+) waves evoked by mechanical stimulation was increased by reduction of extracellular Ca(2+) ([Ca(2+)](o)) or Mg(2+) concentration ([Mg(2+)](o)) and was decreased by elevated [Mg(2+)](o). Reduction of extracellular Ca(2+) concentration ([Ca(2+)](o)) evokes intercellular Ca(2+) signaling in astrocytes; a similar effect was observed in response to change from 5 mM [Mg(2+)](o) to 0 [Mg(2+)](o). Release of low-molecular-weight dyes and ATP was also activated by low [Ca(2+)](o) or [Mg(2+)](o) and inhibited by high [Ca(2+)](o) or [Mg(2+)](o). Astrocytes showed low [Ca(2+)](o)-activated whole cell currents consistent with currents through connexin hemichannels. These currents were inhibited by extracellular Mg(2+). We conclude that extracellular divalent cations modulate intercellular Ca(2+) signaling in astrocytes by modulating the release of ATP, possibly via connexin hemichannels. PMID- 12898706 TI - Microglia density decreases with age in a mouse model of Huntington's disease. AB - Huntington's disease (HD) is characterized by selective neuronal loss and reactive gliosis. In the R6/2 transgenic HD mouse model, there is no selective cell loss, although astrocytosis has been reported. Since there have been no previous studies on microglia in this model, we have undertaken a detailed investigation of microglia in six different forebrain regions in the R6/2 mouse and their wild-type littermates at two time points. Microglia were identified using the histochemical marker isolectin B4 and interactions of genotype, region, and age were analyzed. Results showed that there was a significant decrease in the number of microglia with age in both wild-type and R6/2 brains, which was more pronounced in the transgenic mouse. There were also morphological changes with age observed in both genotypes. As early as 7 weeks of age, structural microglial abnormalities could be seen in R6/2 brains, including bulbous swellings and long stringy processes; comparable changes were seen at 16 weeks in wild-type brains. At 14.5 weeks, microglia in R6/2 mouse brains were smaller in size with condensed nuclei and fragmentation of their processes. We suggest that the density and morphology of microglia change with normal aging and that this process is accelerated in R6/2 brains. Such changes in the dynamic status of microglia may lead to an impairment of their neurosupportive functions. Further studies are needed to understand better the role of microglia in aging and neurodegeneration. PMID- 12898707 TI - Cultured astrocytes express toll-like receptors for bacterial products. AB - It has become apparent that astrocytes may be important contributors to inflammatory immune responses within the brain in response to microbial challenges. To date, the mechanisms that underlie activation of this major glial cell type by such challenges have not been investigated. In the present study, we present evidence for members of a recently discovered family of receptors for highly conserved microbial components, the Toll-like receptors (TLRs), in isolated cultures of primary murine astrocytes. We describe the low-level constitutive expression of messenger RNA-encoding TLR2, TLR4, TLR5, and TLR9 in resting cultures of these cells. Importantly, the level of expression of messenger RNA for each of these receptors is markedly elevated following exposure to specific bacteria-derived ligands for these receptors. The functional expression of these receptor proteins is further supported by the ability of known ligands for each TLR to induce both message expression and protein secretion of the proinflammatory cytokine, interleukin-6. In addition, the recent availability of antibodies to TLR2 and TLR4 has enabled us to demonstrate directly the presence of these receptors on astrocytes by Western blot and immunofluorescence analysis, respectively. Furthermore, we have confirmed the sensitivity of such receptor expression to ligand stimulation. The present demonstration of Toll-like microbial pattern-recognition receptors on primary astrocytes provides a mechanistic link between bacterial challenge and inflammatory immune responses that may be an important component of the pathologies of bacterially induced inflammatory CNS disorders. PMID- 12898708 TI - Binding of an antibody against a noncompact myelin protein to presumptive glial cells in the visual system of the crab Ucides cordatus. AB - Glial cells, in both vertebrate and invertebrate nervous systems, provide an essential environment for developmental, supportive, and physiological functions. However, information on glial cells themselves and on glial cell markers, with the exception of those of Drosophila and other insects, is not abundant in invertebrate organisms. A common ultrastructural feature of invertebrate nervous systems is that layers of glial cell cytoplasm-rich processes ensheath axons and neuronal and glial somata. In the present study, we have examined the binding of a monoclonal antibody to 2',3'-cyclic nucleotide 3'-phosphodiesterase (CNPase) in the compound eye and optic lobe of the crab Ucides cordatus using both light and electron microscopy. CNPase is a noncompact myelin protein that is a phenotypic marker of oligodendroglial and Schwann cells, is apparently involved in the ensheathment step prior to myelin compaction, and is also expressed by the potentially myelinating olfactory ensheathing glia. CNPase has raised much interest, first by virtue of its unusual enzymatic activity and more recently by its membrane-skeletal features and possible involvement in migration or expansion of membranes. We have found CNPase-like immunoreactivity in most cells of the compound eye basement membrane and both in optic cartridges of the synaptic layer and cells of the outer sublayer of the lamina ganglionaris. The results suggest that in the crab visual system some, but not all, glial cells, including some adaxonal glia, may express the noncompact myelin protein CNPase or a related protein. PMID- 12898709 TI - Protective roles of mitochondrial manganese-containing superoxide dismutase against various stresses in Candida albicans. AB - Candida albicans contains copper- and zinc-containing superoxide dismutase but also two manganese-containing superoxide dismutases (MnSODs), one in the cytosol and the other in the mitochondria. Among these, the SOD2 gene encoding mitochondrial MnSOD was disrupted and overexpressed to investigate its roles in C. albicans. The null mutant lacking mitochondrial MnSOD was more sensitive than wild-type cells to various stresses, such as redox-cycling agents, heating, ethanol, high concentration of sodium or potassium and 99.9% O2. Interestingly, the sod2/sod2 mutant was rather more resistant to lithium and diamide than the wild-type, whereas overexpression of SOD2 increased susceptibility of C. albicans to these compounds. The inverse effect of mitochondrial MnSOD on lithium toxicity was relieved when the sod2/sod2 and SOD2-overexpressing cells were grown on the synthetic dextrose medium containing sulphur compounds such as methionine, cysteine, glutathione or sulphite, indicating that mitochondrial MnSOD may affect lithium toxicity through sulphur metabolism. Moreover, disruption or overexpression of SOD2 increased or decreased glutathione reductase activity and cyanide-resistant respiration by alternative oxidase, respectively. Taken together, these findings suggest that mitochondrial MnSOD is important for stress responses, lithium toxicity and cyanide-resistant respiration of C. albicans. PMID- 12898710 TI - Studies on the ATP3 gene of Saccharomyces cerevisiae: presence of two closely linked copies, ATP3a and ATP3b, on the right arm of chromosome II. AB - In this paper, we present evidence that there are two closely linked copies of the ATP3 gene coding for the gamma subunit of the F(1)F(0)-ATPase complex (EC3.6.1.34) in four laboratory strains of Saccharomyces cerevisiae, even though the yeast genome project has reported that ATP3 is a single-copy gene on chromosome II. We previously reported that the gene dosage (three copies) of ATP1 and ATP2 is coincident with the subunit number of F(1)-alpha and F(1)-beta, but that the gene dosage of ATP3 was not consistent with the subunit stoichiometry of F(1)F(0)-ATPase. By applying long PCR and gene walking analyses, we estimated that the two copies of ATP3 were approximately 20 kb apart, and we designated that which is proximal to the centromere ATP3a, while we named that which is distal ATP3b. The nucleotide sequences of the two copies of ATP3 were identical to the reported sequence in the W303-1A, W303-1B and LL20 strains, while only the DC5 strain had a single base substitution in its ATP3a. With the exception of this substitution, the other nucleotide sequences were identical to the upstream 860 bp and the downstream 150 bp. The differences between ATP3 with the single base substitution (Ser(308) to Phe) and ATP3 without the substitution on the complementation of the ATP3 disruptant and on the maintenance of the mitochondrial DNA were observed, suggesting that Atp3ap and Atp3bp in the DC5 strain might have different functions. However, it should not always be necessary for yeast cells to carry different types of ATP3 because the other three strains carry the same type of ATP3. It was also demonstrated that the disruption of the ATP3 genes basically leads to a loss of wild-type mtDNA, but the stability of the mtDNA is not dependent on the ATP3 alone. PMID- 12898711 TI - NET1 and HFI1 genes of yeast mediate both chromosome maintenance and mitochondrial rho(-) mutagenesis. AB - An increase in the mitochondrial rho(-) mutagenesis is a well-known response of yeast cells to mutations in numerous nuclear genes as well as to various kinds of stress. Despite extensive studies for several decades, the biological significance of this response is still not fully understood. The genetic approach to solving this enigma includes a study of genes that are required for the high incidence of spontaneous rho(-) mutants. We have obtained mutations of a few nuclear genes of that sort and found that mutations in certain genes, including CDC28, the central cell-cycle regulation gene, result in a decrease in spontaneous rho(-) mutability and simultaneously affect the maintenance of the yeast chromosomes and plasmids. Two more genes resembling CDC28 in this respect are identified in the present work as a result of the characterization of four new mutants. These two genes are NET1 and HFI1 which mediate important regulatory protein-protein interactions in the yeast cell. The effects of four mutations, including net1-srm and hfi1-srm, on the maintenance of the yeast mitochondrial genome, chromosomes and plasmids, as well as on the cell's sensitivity to ionizing radiation, are also described. The data presented suggest that the pleiotropic srm mutations determining coordinate changes in the fidelity of mitotic transmission of chromosomes, plasmids and mtDNA molecules identify genes that most probably operate high up in the hierarchy of the general genetic regulation of yeast. PMID- 12898712 TI - Functional analysis of the cysteine residues and the repetitive sequence of Saccharomyces cerevisiae Pir4/Cis3: the repetitive sequence is needed for binding to the cell wall beta-1,3-glucan. AB - Identification of PIR/CIS3 gene was carried out by amino-terminal sequencing of a protein band released by beta-mercaptoethanol (beta-ME) from S. cerevisiae mnn9 cell walls. The protein was released also by digestion with beta-1,3-glucanases (laminarinase or zymolyase) or by mild alkaline solutions. Deletion of the two carboxyterminal Cys residues (Cys(214)-12aa-Cys(227)-COOH), reduced but did not eliminate incorporation of Pir4 (protein with internal repeats) by disulphide bridges. Similarly, site-directed mutation of two other cysteine amino acids (Cys(130)Ser or Cys(197)Ser) failed to block incorporation of Pir4; the second mutation produced the appearance of Kex2-unprocessed Pir4. Therefore, it seems that deletion or mutation of individual cysteine molecules does not seem enough to inhibit incorporation of Pir4 by disulphide bridges. In fks1Delta and gsc2/fks2Delta cells, defective in beta-1,3-glucan synthesis, modification of the protein pattern found in the supernatant of the growth medium, as well as the material released by beta-ME or laminarinase, was evident. However, incorporation of Pir4 by both disulphide bridges and to the beta-1,3-glucan of the cell wall continued. Deletion of the repetitive sequence (QIGDGQVQA) resulted in the secretion and incorporation by disulphide bridges of Pir4 in reduced amounts together with substantial quantities of the Kex2-unprocessed Pir4 form. Pir4 failed to be incorporated in alkali-sensitive linkages involving beta-1,3-glucan when the first repetitive sequence was deleted. Therefore, this suggests that this sequence is needed in binding Pir4 to the beta-1,3-glucan. PMID- 12898713 TI - New 'marker swap' plasmids for converting selectable markers on budding yeast gene disruptions and plasmids. AB - Marker swap plasmids can be used to change markers for genes disrupted with nutritional markers in the yeast Saccharomyces cerevisiae. We describe 18 new marker swap plasmids, and we also review other plasmids available for marker conversions. All of these plasmids have long regions of flanking sequence identity, and thus the efficiency of homologous recombination mediated by marker conversion is very high. Marker swaps allow one to easily perform crosses to construct double mutant strains even if each of the disrupted strains contains the same marker, as is the case with the KanMX marker used in the yeast knockout collection. Marker swaps can also be used to change the selectable marker on plasmids, eliminating the need for subcloning. PMID- 12898714 TI - Saccharomyces cerevisiae QNS1 codes for NAD(+) synthetase that is functionally conserved in mammals. AB - NAD(+), an essential molecule involved in a variety of cellular processes, is synthesized through de novo and salvage pathways. NAD(+) synthetase catalyses the final step in both pathways. Here we show that this enzyme is encoded by the QNS1 gene in Saccharomyces cerevisiae. Expression of Escherichia coli or Bacillus subtilis NAD(+) synthetases was able to suppress the lethality of a qns1 deletion, while a B. subtilis NAD(+) synthetase mutant with lowered catalytic activity was not. Overexpression of QNS1 tagged with HA led to elevated levels of NAD(+) synthetase activity in yeast extracts, and this activity can be recovered by immunoprecipitation using anti-HA antibody. An allele of QNS1 was constructed that carries a point mutation predicted to reduce the catalytic activity. Overexpression of this allele, qns1(G521E), failed to elevate NAD(+) synthetase levels and qns1(G521E) could not rescue the lethality caused by the depletion of Qns1p. These results demonstrate that NAD(+) synthetase activity is essential for cell viability. A GFP-tagged version of Qns1p displayed a diffuse localization in both the nucleus and the cytosol. Finally, the rat homologue of QNS1 was cloned and shown to functionally replace yeast QNS1, indicating that NAD(+) synthetase is functionally conserved from bacteria to yeast and mammals. PMID- 12898715 TI - The ailing and failing implant: prevention and treatment. PMID- 12898716 TI - [Pars plana vitrectomy and primary implantation of silicone oil in the treatment of acute exogenous endophthalmitis in eyes without retinal detachment]. AB - Acute exogenous endophthalmitis (AEE) is the most serious post-traumatic or postoperative complication which without immediate treatment leads to amaurosis or anatomical loss of the eye. If comprehensive antibiotic treatment fails immediate pars plana vitrectomy (PPV) is indicated. Empirically and later also by laboratory methods the antiinflammatory effect of silicone oil (SO) was revealed. The objective of the present retrospective study is to demonstrate anatomical and functional results of PPV with implantation of SO for AEE on eyes without detachment of the retina. In 1990-2000 the authors operated a total of 22 eyes of 22 patients with AEE. The mean age of the patients was 59.6 years, the mean observation period 35.5 months. In 17 eyes postoperative and in 5 eyes post traumatic AEE was involved. The mean interval between the original operation and PPV was 20.8 days and between the injury and PPV 5.4 days. In all patients we took at the beginning of PPV specimens of the vitreous body for cultivatin which gave in 63.6% a positive result. The intraocular lens was explanted in 81.8% eyes. After subtotal resection of the vitreous body we implanted SO and in all aphakic eyes we performed iridectomy in no. 6. PPV was completed by administration of vancomycin and ceftazidime into the vitreous space. In 95.4% of the eyes we preserved anatomically the bulbus, in one instance we eviscerated the bulbus three days after failure of PPV (4.6%). In 86.4% the retina remained anatomically attached and in the remaining 9% eyes beneath the SO traction detachment of the retina developed. Visual acuity improved in 72.7% eyes, it remained unchanged in 9.1% eyes and deteriorated in 18.2% eyes. Silicone oil was explanted in 40.9% eyes after a mean interval of 4.6 months following PPV. In the remaining patients we did not indicate explantation of SO, in particular because of the patients' advanced age, CONCLUSION: We recommend PPV and implantation of SO in AEE on eyes without detachment of the retina as the method of choice. PMID- 12898717 TI - [Pneumatic retinopexy--method of choice in treatment of retinal detachment]. AB - Authors are presenting their experience with pneumatic retinopexy (PR) in the therapy of primary rhegmatogenous retinal detachment. Technique was for the first time described by Hilton and Grizzard in 1986 as an intravitreal injection of expending gas in combination with transconjunctival cryotherapy or laser photocoagulation of retinal tears. After gas application a patient, particularly his head, must be positioned the way that the rising gas bubble perfectly seals the retinal tear. This also subretinal fluid to be reabsorbed. In the study group of 30 eyes, 11 were pseudophakic and 19 phakic. The reattachment after single PR operation was 77% (23 eyes), in 24% (7 eyes) of cases reoperation was needed. 2 cases required multiple injection of the gas, in 5 cases retinal reattachment after PR was never occurred. The most common causes of failure to reattach the retina were initiation and acceleration of proliferative vitreoretinopathy. In these cases a following operation was needed--most frequently pars plana vitrectomy with tamponading the vitreous cavity by gas or silicone oil. The final reattachment rate was then 94% (28 eyes), one patient refused reoperation, in one case retina remained detached. Pneumatic retinopexy is quite modest and time saving operative technique which can be used in indicated cases as a one-day surgical procedure, which corresponds with current heading of the eye microsurgery. Although there is always the possibility of failure to reattach the retina using PR as a method of choice. The risk of failure can be significantly lowered by adequate selection of patients. PMID- 12898718 TI - [Importance of mathematical analysis of the retinal image in diabetic retinopathy]. AB - The authors test whether for evaluation of the severity of diabetic retinopathy it is possible to use the method of computer assisted mathematical processing of the retinal image. This method is based on the identification of vascular endings in the visualized part of the retina. It is assumed that with the severity of the finding the number of vascular endings increases in conjunction with the process of neogenesis. For assessment of the number of vascular endings the authors use the Adaptive Contrast Control (ACC) method. By a special procedure the vascular endings are identified and their number is assessed in the investigated area of the retina. The method makes it possible to assess in addition to the number of vascular endings also the total length of the vessels, their volume, surface, area they cover and the histogram of the diameter of the vessels. The authors examined 19 patients (38 eyes) with a quite normal finding on the retina. The mean age of the group was 37.3 years. Moreover they examined 10 patients (20 eyes) with middle advanced and advanced non-proliferative diabetic retinopathy and 10 patients (20 eyes) with risk proliferative diabetic retinopathy. In every patient a standard digital photograph of the fundus of both eyes size "tif" was made and each picture was subjected to mathematical analysis. The group of patients was divided into three sub-groups (patients with a normal finding, a non proliferative finding and a finding of proliferative diabetic retinopathy). In the conclusion the authors provide evidence that with increasing severity of the finding on the retina also the number of vascular endings increases. PMID- 12898719 TI - [Immunofluorescence identification of limbal stem cells in a culture of donor corneas]. AB - The purpose of this study was to identify limbal stem cells in the culture of human donor cornea. After standard cultivation procedure we used monoclonal antibody against alpha-enolase for identification of stem cells. This antibody serves as a probe for the identification of biochemical marker of stem cells. Using immunofluorescent microscopy we identified in specimens alpha-enolase positive cells which in vitro but not in situ showed also some morphologic differences from other cellular elements of the culture. PMID- 12898720 TI - [Microbial inflammations of the orbit in children]. AB - The author evaluates in a retrospective study the incidence, diagnosis and causal agent and therapeutic results of microbial inflammations of the orbit. During the period from Jan. 1 1997 till Dec. 31 1999 66 children were hospitalized with the diagnosis of inflammation of the orbit. The mean age of the children was 6.2 years, the mean period of hospitalization 10.2 days. Chandler's classification of inflammations of the orbit was used. The diagnosis was established in collaboration by an ophthalmologist, otolyryngologist and roentgenologist. The main cause of inflammations of the orbit was inflammation of the paranasal sinuses. The most frequent microbial agents were streptococci, staphylococci and Haemophilus influenzae. All patients were treated during hospitalization by antibiotics administered by the i.v. route. Surgery was necessary in 8 patients- in 4 patients with an orbital abscess and in 4 patients with a subperiostal abscess not responding to conservative treatment. The operation was performed by an otolaryngologist using always the endonasal approach. The authors did not record in their group any permanent ophthalmological complications resulting from the inflammation of the orbit. PMID- 12898721 TI - [Effect of strabismus on the quality of life in adults]. AB - The aim of this paper was to prove by help of questionnaire the psychosocial problems in 26 adult patients operated for strabismus. We stated that the biggest difficulties of patients occurred by work and sport activities. The patients with diplopia also complained of difficulties with uncertain orientation, walking on steps and by locating the objects. The uncertainty by communication was also a great problem. The surgery fulfilled the expectations by 18 patients (69%), 8 patients (31%) were disappointed. PMID- 12898722 TI - [Corneal surface analysis]. AB - Fourier analysis is a powerful method of evaluating the surface of the cornea. In 50 patients after photorefractive keratectomy included in our study we measured spherical equivalent, regular astigmatism, irregular astigmatism and decentration retrospectively acquired by Fourier series analysis of corneal topography data 1, 3, 6 and 12 months after operation. Postoperative decentration increased significantly from a mean preoperative value. The preoperative and postoperative values are not significantly different. The Fourier spherical equivalent and the values of keratometric spherical (equivalent) are highly correlated. Correlation among decentration, irregular astigmatism and best corrected visual acuity is important for clinical practice. Due to Fourier analysis we obtain information, which could explain worse results of some surgeries. PMID- 12898723 TI - [Refractive surgery in myopia up to -6D]. AB - OBJECTIVE: To evaluate the clinical results in patients with myopia up to -6 dioptres (D) after refraction surgery, evaluate their stability and compare photorefractive keratectomy (PRK) and laser in situ keratomileusis (LASIK). MATERIAL AND METHODS: The authors evaluated before and after surgery the refraction values in patients with myopia up to -6 D operated in 1999 and 2000. The patients were subjected to standard examination before operation, one week, 1, 3, 6 and 12 months after operation. The group comprised a total of 542 eyes, 452 PRK eyes and 90 LASIK eyes. RESULTS: After PRK surgery the mean spherical refraction after one year was -0.35 +/- 0.71 D, the mean astigmatism -0.63 +/- 0.38 D, UCVA 0.90 +/- 0.19, BCVA 0.98 +/- 0.11. After LASIK surgery after one year the mean spherical refraction was -0.10 +/- 0.81. D, the mean astigmatism 1.05 +/- 0.65 D, UCVA 0.80 +/- 0.24, BCVA 0.86 +/- 0.21. CONCLUSION: Both methods proved effective and safe for correction of myopia up to -6 D. The results at our clinic are comparable with other departments and are stable in different years. PRK and LASIK achieve equal postoperative results after an interval of 3 to 6 months following surgery. PMID- 12898724 TI - [Long-term results of photorefractive keratectomy in myopia up to -6 D]. AB - OBJECTIVE: To present clinical results of patients with myopia up to -6 dioptres (D) after photorefractive keratectomy (PRK) and evaluate their stability during a 4-year period. MATERIAL AND METHODS: The authors evaluated retrospectively the refraction before and after surgery in patients with myopia up to -6 D during the period from September 1997 till December 2000. The group comprised a total of 847 eyes. The mean spherical refraction during the whole follow up period was before surgery -3.61 +/- 1.17 D, the mean astigmatism was -0.65 +/- 0.63 D, non corrected visual acuity (UCVA) 0.09 +/- 0.09, the visual acuity with optimal correction (BCVA) 1.21 +/- 0.30. RESULTS: One year after PRK we achieved in our group a mean spherical refraction of -0.17 +/- 0.74 D, a mean astigmatism of 0.61 +/- 0.43 D, UCVA 0.92 +/- 0.25, BCVA 1.02 +/- 0.19. CONCLUSION: Photorefractive keratectomy is an effective and safe method for correction of myopia up to -6 D. The results as regards refraction were comparable with other departments and did not differ much during individual years. Stabilization occurs 3-6 months after the operation. PMID- 12898725 TI - [Chirocaine in ophthalmology]. AB - The author presents an account on the properties and initial own experience with a new long acting amide local anaesthetic levobupivacaine. Levobupivacaine supplied under the name Chirocaine is a laevorotatory enanciomere of racemic bupivacaine known this country under the name Marcaine. Contrary to racemic bupivacaine levobupivacaine has a lower cardiotoxicity and neurotoxicity while it has the same anaesthetic and analgetic effect. The author tested the anaesthetic in 100 patients using the same dosage as in hitherto commonly used bupivacaine. Based on data in the literature and their own experience the authors consider levobupivacaine very suitable also for ophthalmosurgery, in particular in risk patients and in long operations where a larger amount of anaesthetic is necessary. PMID- 12898726 TI - [Simulation of quality of vision in a woman accused of killing her husband with a firearm--comments from a forensic expert]. PMID- 12898727 TI - [Compendium of the first 100 years of scientific articles, writings, documents, editorials, and historical facts about medicine in the light of the times of the Boletin Cientifico de la Asociacion Medica de Puerto Rico]. PMID- 12898728 TI - [Uncinariasis. 1916]. PMID- 12898729 TI - [Apropos of the current outbreak of smallpox. 1916]. PMID- 12898730 TI - [Tuberculosis in Puerto Rico. What has been done to combat it; what is being done, what remains to be done: modern approaches. 1926]. PMID- 12898731 TI - [Inauguration of the chair of the Asociacion Medica de Puerto Rico. Address of President Dr. Ramon M. Suarez]. PMID- 12898732 TI - [Malaria in Puerto Rico. 1931]. PMID- 12898733 TI - [The campaign against cancer. Extent, volume, and practical result of work performed during the 1st year of the emergency clinical of the Puerto Rican League against Cancer]. PMID- 12898734 TI - The position of the University of Puerto Rico. 1947. PMID- 12898735 TI - [Practical points in the treatment of the diabetic patient. 1956]. PMID- 12898736 TI - An epidemiological study on coronary heart disease in PR. The Puerto Rico Heart Health Program. 1969. PMID- 12898737 TI - [Pain in the lower back. Etiological factors and clinical evaluation. 1971-1980]. PMID- 12898738 TI - Bronchial asthma in Puerto Rican children. 1971-1980. PMID- 12898739 TI - [The ground-breaking ceremony of the Cardiovascular Center of Puerto Rico and the Caribbean. 1987]. PMID- 12898740 TI - [Depression and anxiety. Etiological factors and clinical evaluation. 1991]. PMID- 12898741 TI - [Dr. Diego Alvarez Chanca. Introducer of universal medicine in America. 1992]. PMID- 12898742 TI - Characteristics and survival of adult AIDS cases in Puerto Rico, 1999. PMID- 12898743 TI - Prenatal diagnosis of right-sided diaphragmatic hernia. 1997. PMID- 12898744 TI - [Our thought. 1903]. PMID- 12898745 TI - [Pediatrics, medical research and the health of the children in Puerto Rico: Pessimism or crude reality?]. PMID- 12898746 TI - Severe neutropenia in an infant with methylmalonic acidemia. AB - Neutropenia exists when the neutrophil counts is less than 1000/mm3 in infants between 2 weeks and 1 year of age and less than 1500/mm3 beyond 1 year of age (1). Severe infections occur when the absolute neutrophil count is below 500/mm3 with perirectal abscesses, pneumonia, and sepsis being common. Granulocyte Colony Stimulating Factor (G-CSF) produces a sustained neutrophil recovery in patients with severe neutropenia, reduces the incidence and severity of infection, and improves the quality of life. Various cytopenias, including neutropenia, thrombocytopenia and pancytopenia, have been reported in association with inborn errors of branched aminoacid metabolism such as methylmalonic, propionic and isovaleric acidemia. We report an infant with methylmalonic acidemia who presented severe neutropenia. PMID- 12898747 TI - Childhood cancer and hematopoietic stem cell transplantation: a seven year experience at University Pediatric Hospital in Puerto Rico. PMID- 12898748 TI - Evaluation of patient age as a risk assigment parameter in high risk acute lymphocytic leukemia. AB - Historically, age has been used as criteria for risk group assignment in patients with acute lymphocytic leukemia (ALL). The prognosis varies among different age groups. Differences in the biological characteristics of the leukemic clone and clinical findings at diagnosis are also used for risk assignment in ALL. It is our hypothesis that age is a lesser risk defining parameter than are the biological characteristics of the leukemic clone and the clinical characteristics at presentation. An analytic, retrospective study was conducted where fifty-seven patients were evaluated. Patients were divided in two subgroups: nine years and older (up to 18 years of age) patients with favorable biological characteristics of the leukemic clone and favorable clinical findings at diagnosis (riesgo 0) and nine years and older (up to 18 years of age) patients with unfavorable characteristics of the leukemic clone and/or unfavorable clinical findings at diagnosis (riesgo 1). All patients received treatment according to the same treatment protocols. The subgroup of patients with favorable ALL characteristics (riesgo-0) showed an increased median survival time of 11 years compared to those patients with unfavorable ALL characteristics (riesgo-1) who had a median survival time of 5.4 years. Age as a risk assignment perameter had a minimal impact in survival in patients whose leukemia had good biologic characteristics treated on these treatment protocols. PMID- 12898749 TI - Technique for automated exchange transfusion in pediatric patients with sickle cell crisis. PMID- 12898750 TI - [Pediatrics graduate medical education in Puerto Rico in the 21th Century]. PMID- 12898751 TI - [Notes for the history of pediatrics in Puerto Rico]. PMID- 12898752 TI - [The effect of gene transfection of vascular endothelial growth factor-C on the metastasis ability of Tca8113]. AB - OBJECTIVE: To study the role that vascular endothelial growth factor-C plays in the lymph node metastasis of the squamous cell carcinoma (SCC) of tongue. METHODS: An established VEGF-C transfected Tca8113 cell strain (VcTca) was heterotransplanted into BALB/C nude mice subcutaneously. The expression of VEGF-C and VEGFR-3 was detected. The property of tumorigenesis and metastasis of the transfected cell was investigated. RESULTS: Jugular lymphatic metastases developed in 4/6 mice of VcTca group in 6 weeks after transplantation, while no lymph node metastasis was found in mice with tumors of non-transfected Tca8113 cell (Tca) and mock transfected cell(pDTca) at the same time. There was no difference in tumor weight among VcTca, pDTca and Tca (P > 0.05). CONCLUSION: It is suggested that VEGF-C could promote lymph node metastatic ability of Tca8113 cell, and could be utilized as a target of new regimen in prevention and treatment for lymphatic metastasis of oral carcinomas. PMID- 12898753 TI - [Construction and expression of traceable DNA vaccine for prevention of caries]. AB - OBJECTIVE: Streptococcus mutans has been proved as a causative bacteria of human dental caries. The surface protein antigen is one of the important pathogenic factors. The A region of the surface protein antigen pac gene (pacA) can enrich T cells and B-cells epitope. In this study, a DNA vaccine carrying pacA and gfp gene (a reporter gene) for caries prevention was constructed. The DNA vaccine was liable to be traced in vitro and in vivo. METHODS: The fragment of pacA (1.3 kb) was amplified by PCR with the plasmid pPC41 as template, and inserted into a pEGFP-C1 vector. The recombinant plasmid produced was named as pEGFPC1-pacA. After the COS1 cell line was transfected by the recombinant plasmid, the expression of gfp was detected by observing the green fluorescence and measuring the fluorescence intensity, and the expression of pacA was detected by RT-PCR. RESULTS: Restricted analyzing, sequencing and PCR technique were employed to identify the recombinant plasmid. The phase and orientation of the pacA gene inserted into the vector pEGFPC1 were correct and no changes of their open reading frames were discovered. The transfected COS1 carrying green fluorescent protein (GFP) was observed; the GFP expression level of transfected cells was higher than that of controlled cell. The transcript of pacA gene was confirmed by RT-PCR. CONCLUSION: Construction of the recombinant plasmid was successful. The gfp gene and pacA gene in the plasmid was transcribed and expressed simultaneously in the transfected cells. Moreover, detection of GFP is simple, safe and effective for living cells. PMID- 12898754 TI - [A study on age-related changes of histological structure and DNA repair capacity in oral mucosa]. AB - OBJECTIVE: To investigate the age-related changes of histological structure and DNA repair capacity (P53 protein expression) in oral mucous membrane and try to explain why the elders are prone to oral squamous cell carcinoma. METHODS: The routine HE staining and an immunohistochemical technical (SP method) were used respectively to study the changes of histological structure and P53 expression in normal oral mucosa of different age groups. The SP staining results were evaluated by semiquantity method. RESULTS: 1. With aging, a series of histological changes were observed in the oral mucosa; 2. With aging, an elevating tendency of P53 protein level emerged in the oral mucosa (P < 0.05). CONCLUSION: The sensitivity to carcinogenic agents in elder people may be accentuated by the age-related histological changes and DNA repair capacity changes of the normal oral mucosa. PMID- 12898755 TI - [The effects of nm23-H1 on metastases ability and chemo-sensitivity in Tca8113 cell lines]. AB - OBJECTIVE: The purpose of this study was to establish a stable, high-efficient and low-toxic way of transfecting nm23-H1 into Tca8113 line cells, and then to find out whether nm23-H1 could affect the invasion and metastases ability of Tca8113 line cells. METHODS: nm23-H1 was transfected into Tca8113 line cells with Lipofect. The different expressions of nm23-H1 between transfected and non transfected line cells were detected by the method of immunohistochemistry. The difference of the invasion and metastases ability between transfected and non transfected line cells was detected by transwell-room and wash techniques. The change of chemo-sensitivity was evaluated by MTT. RESULTS: Using pCMV-NEO-BAM system to keep stable expression of nm23-H1, the significant difference of NDPKA expression between transfected and non-transfected Tca8113 line cells was discovered; The metastases ability of transfected Tca8113 line cells decreased significantly; The chemo-sensitivity of transfected Tca8113 line cells to CDDP increased significantly. CONCLUSION: nm23-H1 can inhibit the metastases of Tca8113 line cells and increase the chemo-sensitivity to CDDP significantly. PMID- 12898756 TI - [The effect of cryotreat on the wear resistance of the medium melting-point castable alloy]. AB - OBJECTIVE: The purpose of this study was to introduce cryotreat technique into prosthetic dentistry by investigating the wear resistance changes of SDA-II medium melting-point castable alloy before and after cryotreat. METHODS: 15 samples were divided into three groups, i.e. control group, cryotreat group, cryotreated plus post-cryogenic treat group. Weight loss measurement was used to examine the wear resistance. RESULTS: For SDA-II medium melting-point castable alloy, cryotreat effectively improved its wearing resistance; so far as the wear resistance was concerned, the post-cryogenic treatment was unnecessary. CONCLUSION: The results obtained above suggest that cryotreat was an effective method in enhancing the wear resistance of SDA-II medium melting-point castable alloy. PMID- 12898757 TI - [The effect of deep branches of cervical nerve on motorial function of trapezius muscle]. AB - OBJECTIVE: The aim of this study was to examine the effects of deep branches of cervical plexus on trapezius muscle. METHODS: The evoked electromyography of 32 trapezius of 16 SD rats was examined in different time and results obtained were processed by statistical methods. RESULTS: After cutting the accessory nerves, stimulating the deep branches of cervical plexus evoked the myoelectricity, the myoelectricity results varied according to different time. CONCLUSION: The deep branches of cervical plexus are another important motorial supply to trapezius muscles. After cutting the accessory nerves, if the branches were undamaged, the shoulder's function would be greatly preserved. PMID- 12898758 TI - [The effects of different enamel etching time on bonding strength]. AB - OBJECTIVE: The aim of this study was to investigate the effect of short enamel etching time on bonding strength. METHODS: 80 healthy premolars were randomly divided into two equal groups, one was etched for 15 seconds, the other for 60 seconds, after normally bonding Edgewise brackets, the tooth specimen was stored in water at room temperature for 24 hours. An MTS NEW810 100KN testing machine was used to examine the shear force. RESULTS: Although the bonding strength for 15 seconds etching time was weaker than that for 60 seconds, the means of the bonding strength in 15 seconds group reached 5.8625 MPa. CONCLUSION: Etching for 15 seconds could provide enough bond strength for orthodontic practice. Furthermore, the adhesive remained on tooth after debonding was less by comparison with 60 seconds etching group, therefore brackets could be removed easily and the work efficiency could be increased in clinic by means of 15 seconds etching time. PMID- 12898759 TI - [Survival rate in mucoepidermoid carcinoma of salivary glands]. AB - OBJECTIVE: To study the clinico pathologic factors influencing postoperative survival rate in mucoepidermoid carcinoma of salivary glands. METHODS: A retrospective study of seventy patients with mucoepidermoid carcinoma of salivary glands has been carried out. All cases underwent primary surgical resection in our hospital were followed up and clinicopathologic factors influencing the postoperative survival rate of patients were investigated. Statistical analysis was conducted using a computer program (SPSS). The data were statistically treated according to the Kaplan-Meier method. The Log rank tests were employed to assess the statistical significance of various groups. RESULTS: The 3-, 5- and 10 year overall survival rate were 90.66%, 88.85% and 86.63%, respectively. The patients with tumors' diameter less than 4 cm, early clinical stage (stage I and II), well differentiated tumor cells, without lymph nodes involvement, without local recurrence and metastasis after primary surgical resection had a long survival term. The patients with tumors' diameter more than 4 cm, with lymph nodes involvement, advanced clinical stage (stage III and IV), poor differentiated, with local recurrence and metastasis after primary surgical resection demonstrated a short survival term. CONCLUSION: TNM and clinical stage, the degree of differentiation, with or without local recurrence and metastasis after primary surgical resection are the important factors influencing the postoperative survival time. PMID- 12898760 TI - [The neck treatment of cN0 patients with squamous cell carcinoma of buccal mucosa]. AB - OBJECTIVE: The purpose of this study was to discuss the principle in neck treatment of cN0 patients with squamous cell carcinoma of buccal mucosa. METHODS: 101 patients of squamous cell carcinoma of buccal mucosa at the stage of cN0, who had hospitalized in West China College of Stomatology, Sichuan University from 1980 to 2000, were investigated retrospectively. All the patients received a comprehensive therapy consisting of surgical procedures combined with chemotherapy and radiotherapy. The combining radical therapy of buccal, mandible and neck was the main surgical method. Lymph nodes were cleared after operation and examined by pathological method. The patients had been followed up for more than 3 years. RESULTS: 17 patients had lymph nodes metastasis, the occult metastasis (OM) rate was 16.83%. It increased in the high stage of original lesion, the OM rate of T3 and T4 was 18.18% and 52.00%, respectively. The metastasized lymph nodes were mainly located in submandible and superior deep cervix lymph nodes, their respective metastasis rate was 41.18% and 29.41%. CONCLUSION: The rate of occult metastasis of squamous cell carcinoma of buccal mucosa is high, and then we should adopt actively selective neck dissection for the cN0 patients of squamous cell carcinoma of buccal mucosa. PMID- 12898761 TI - [Clinical study on chitosan in curing irreducible anterior disc displacement of tempormandibular joint]. AB - OBJECTIVE: To evaluate the clinical efficacy of intra-articular injection with 2% chitosan as a treatment for irreducible anterior disc displacement of the TMJ. METHODS: Total 30 patients with irreducible anterior disc displacement were divided into two groups according to quasi-randomizaion. The patients in test group received intra-articular injection with 1.0 ml of 2% chitosan into upper cavities of the suffered joints, in control group with 12.5 mg of prednisolone. The patients were followed up at the 1st day and 14th day after injection and the maximal mouth opening was measured. RESULTS: The maximal mouth opening at 14th day was (36.73 +/- 4.69) mm in test group, with 11.73 mm increase from baseline; and (28.53 +/- 5.81) mm in control group, with 3.86 mm increase, respectively. The increase of maximal mouth opening in test group was significantly higher than that of control group (P < 0.05). CONCLUSION: Chitosan is an effective biomaterial in curing irreducible anterior disc displacement of TMJs. PMID- 12898762 TI - [The study on the clinical manifestation of developmental enamel defects in primary dentition]. AB - OBJECTIVE: To study the clinical manifestation and its pathogenesis of the developmental enamel defects in primary dentition of children with low birth weight and premature birth history. METHODS: One hundred and seventy-six children (aged 3-8 years old) were studied about the clinical manifestation of the developmental enamel defects in the primary dentition and its relationship with their medical history. RESULTS: The prevalence of enamel defects in primary dentition in these children was 77.3%. There was no significant correlation between enamel defects and gender. Enamel opacity mostly affected the upper and lower second primary molars. Enamel hypoplasia mostly affected the maxillary and mandibular primary incisors and the maxillary first primary molars. CONCLUSION: Enamel defects mainly result from children's general disorder at birth or within one year after birth. PMID- 12898763 TI - [The effect of root canal preparation with nickel-titanium rotary instruments in reducing post-operative pain]. AB - OBJECTIVE: To examine the effect of nickel-titanium rotary instruments in reducing post-operative pain. METHODS: 95 molars with pulpal and/or periapical involvement were randomly assigned to two groups for root canal preparation. One group was treated with ProFile nickel-titanium rotary instruments; the other was treated by using stainless steel hand files-FlexoFile. The incidence and degree of post-operative pain after root canal preparation in the two groups were recorded. RESULTS: In ProFile group, the incidence of post-operative pain was 27.7%, while 62.5% of the teeth developed post-operative pain after hand preparation. The difference between two groups was statistically significant (P < 0.01). CONCLUSION: Root canal preparation on molars with stainless steel hand files resulted in relatively high incidence of post-operative pain. Mechanical preparation using ProFile nickel-titanium instruments generated less post operative pain and discomfort. PMID- 12898764 TI - [Comparison of extraction versus non-extraction orthodontic treatment results--a preliminary study]. AB - OBJECTIVE: The purpose of this study was to make a comparison between extraction and non-extraction orthodontic treatment results concerning teeth alignment, occlusal relation, facial profile and treatment interval. METHODS: 39 extraction non-extraction borderline cases were selected by 5 orthodontic specialists through reviewing their pre-treatment records. 5 specialists were then asked to rank the results by marking the post-treatment record of each case. And then the samples were divided into different groups according to extraction, non extraction and extraction pattern by reviewing case history. RESULTS: No statistically significant difference was found in teeth alignment, overbite and overjet, midline symmetry, lateral occlusal relation and treatment intervals. However, the post-treatment facial profile of extraction groups obtained higher marks from 5 specialists (P = 0.001). CONCLUSION: While both extraction and non extraction treatment can achieve comparable results in teeth alignment and occlusal relation with equivalent time interval in borderline cases, extraction treatment, whatever extraction of four first premolars or second premolars is performed, can achieve better facial profile. PMID- 12898765 TI - [Establishment of normal value of posterior arch length of adolescence in Chengdu]. AB - OBJECTIVE: To investigate the development of third molar embryo and establish the normal value of posterior arch length of adolescence in Chengdu. METHODS: The samples consisted of 40 males and 41 females with Class 1 dentitions, normal second molar occlusion, no history of orthodontic treatment, and good facial balance. Cephalometry and statistic analysis were conducted for all the subject. RESULTS: The normal value of posterior arch length of maxilla in male was(16.52 +/- 2.35) mm and in female was(16.42 +/- 2.55) mm. The normal length of mandible was(13.00 +/- 2.44) mm in male and(12.43 +/- 2.18) mm in female. But statistic analysis showed no gender difference. Most of the third molar embryo had been formed and their tooth crowns had been calcified, the occurrence ratio was 84.0% in maxilla and 85.2% in mandible. Most of them located in the middle and inclined mesially. CONCLUSION: The data obtained in this study will be valuable for our clinical practice and be helpful for the study concerning dento-facial growth and development. PMID- 12898766 TI - [A study of Bolton tooth-size discrepancies of malocclusion patients]. AB - OBJECTIVE: To analysis the sum and frequencies of Bolton tooth-size discrepancies Angle Class I, II, III malocclusion patients. METHODS: Measured each tooth crown mesial-distal size between the first molar of 439 dental plaster casts of malocclusion patients and obtained the sums of six anterior tooth-size and twelve tooth-size between the first molar of upper or lower and calculated the anterior and total tooth-size discrepancies by Bolton rate standards, then statistic analyses were done. RESULTS: Patients whose sums of anterior tooth-size discrepancy were pass 1.5 mm or less than -1.5 mm was 14.02% in Angle Class I malocclusion patients and 9.49% in Class II and 19.32% in Class III. Patients whose sums of total tooth-size discrepancy were pass 1.5 mm or less than -1.5 mm was 19.63% in Angle Class I malocclusion patients and 15.33% in Class II and 20.45% in Class III. The upper sum of anterior tooth-size of Class I malocclusion patients whose sums of anterior tooth-size discrepancies were pass 1.5 mm or less than -1.5 mm was always smaller than normal and the lower sum was always larger. The sums of anterior tooth-size discrepancy of Class I, II, III and total tooth size discrepancy of Class I were always between 1.5 mm-2.5 mm or -2.5 mm(-)-1.5 mm. Patients whose sums of anterior tooth-size discrepancy were pass 3.5 mm or less than -3.5 mm was 2.34% in Class I malocclusion patients and 0 in class II and 0 in class III. Patients whose sums of total tooth-size discrepancy were pass 3.5 mm or less than -3.5 mm was 4.21% in Class I and 1.46% in Class II and 4.54% in Class III. CONCLUSION: Tooth-size discrepancy of malocclusion patients was not the general cause of malocclusion. PMID- 12898767 TI - [The influences of metal-ceramic restoration margins on gingival health]. AB - OBJECTIVE: Today metal-ceramic restorations are widely accepted in dental practice because of their good aesthetic and biocompatible properties, but there are still several problems to be resolved, among which the influences of their margins on gingival health is noticeable. The purpose of this study was to investigate the influences of metal-ceramic restoration margin on gingival health. METHODS: 129 abutments of metal-ceramic restorations, including their margins' fitness, location and smoothness were examined and their influences on gingival health were analyzed. RESULTS: There was a close relation between the metal-ceramic restorations' margins and the gingival health. CONCLUSION: In order to maintain the gingival health, the margins should be smooth, closely fitted, no overhanging and in a moderate location (i.e. not below the gingival more than 1 mm). PMID- 12898768 TI - [A study on the color difference between Au-Pt alloy porcelain and Ni-Cr alloy porcelain]. AB - OBJECTIVE: To investigate the color difference between Au-Pt alloy porcelain and Ni-Cr alloy porcelain. METHODS: 30 metal-ceramic specimens with different dentin porcelain thickness were fabricated with two types of metal-ceramic alloy, each type of alloy had 15 specimens. L*, a*, b* were measured after opaque porcelain was applied, and dentin porcelain was fired 1, 3, 5, 7 times by MINOLTA CR-100. Then delta E was calculated which reflected the color difference between high gold alloy porcelain and Ni-Cr alloy porcelain. RESULTS: Comparing with Ni-Cr alloy porcelain, the color of Au-Pt alloy porcelain was reddish, yellowish and less bright. The delta E between high-gold alloy porcelain and Ni-Cr alloy porcelain in shade A2 was largest when opaque porcelain was applied. It decreased when dentin porcelain was applied. It became smallest when fired 3 times, and increased along with the increase of fire times. It was larger than 1.5 except firing 3 times. When dentin porcelain was applied, delta E which was larger than 1.5 among different dentin porcelain thickness decreased along with the increase of dentin porcelain thickness. CONCLUSION: The color difference between the two types of metal-ceramic alloy should be carefully taken into account in order to improve the quality of color matching. PMID- 12898769 TI - [Clinical evaluation on effects of functional muscular equal pressure zone on complete denture]. AB - OBJECTIVE: To study the clinical effect of the functional muscular equal pressure zone on complete denture. METHODS: Artificial teeth were arranged within the functional muscular equal pressure zone. With this technique 35 complete dentures were made to rehabilitate 35 edentulous patients whose alveolar bones were moderately or seriously absorbed. After three months' following up, the satisfaction of these patients were investigate. RESULTS: After prosthesis treatment, 35 patients enunciated well and the aesthetic effect was favorable. 89 percent of these patients satisfied the retention and the stability of the maxillary denture, and 78 percent satisfied the retention and the stability of the mandibular denture. 79.5 percent of these patients satisfied the mastication efficiency. CONCLUSION: With the technique, the complete denture is fairly stable in patient's mouth. The technique can enhance the denture's retention and improve the patient's mastication efficiency, especially to these whose alveolar bones are moderately or seriously absorbed. Clinically the technique is of high practical value to these edentulous patients who are difficult to be rehabilitated. PMID- 12898770 TI - [Clinical evaluation of two temporary restoration materials: Composite and self cured acrylic resin]. AB - OBJECTIVE: The purpose of this study was to compare the clinical application and effect of two temporary restorations materials: composite and self-cured acrylic resin. METHODS: 153 composite temporary crowns (and bridges) placed in 44 patients and 144 self-cured acrylic resin temporary crowns (and bridges) placed in 49 patients were evaluated according to the California Dental Association's (CDA) criteria and questionnaire. RESULTS: It showed that composite temporary crowns (and bridges) had better surface smoothness, marginal integrity and anatomic form than self-cured acrylic resin temporary crowns (and bridges). In the 49 patients with self-cured acrylic resin temporary crowns (and bridges), 67% and 88% of them complained of food impaction and mouth odor, while only 2% and 30% in the 44 patients with composite temporary crowns (and bridges) had the same complaint. CONCLUSION: Between the two temporary restoration materials, composite is better than self-cured acrylic resin. PMID- 12898771 TI - [The study of malocclusion of treatment priority index in Xi'an adolescent]. AB - OBJECTIVE: To evaluate the prevalence of malocclusion and orthodontic treatment priority in adolescent. METHODS: 591 boys and 567 girls (aged 12 years old) were studied with their study models and the treatment priority index (TPI) were obtained. RESULTS: 7.03% adolescent had orthodontic treatments (TPI 1:21.33%, TPI 2:47.06%, TPI 3-TPI 5:31.61%). CONCLUSION: TPI is a valid tool to evaluate malocclusion and orthodontic treatment priority. It's very important to establish a regular examination system need for orthodontic treatment in school. PMID- 12898772 TI - [Study on potential anti-caries DNA vaccine pcDNA3-gtfB integration into host cell genome]. AB - OBJECTIVE: Gene vaccine security is of concern because of the possibility of insertion mutagenesis resulting in inactivation of tumor suppressor or activation of oncogene. The purpose of this study was to examine the potential of anti caries DNA vaccine pcDNA3-gtfB integrating into the host cell genome. METHODS: Anti-caries DNA vaccine pcDNA3-gtfB was constructed by the previous study. The gtfB gene(904-4,578 bp, genebank M17361) was cloned from Streptococcus mutans GS 5. 36 Wistar rats were divided into 2 groups: submandibular gland-targeted injection(SGT) group and control group. Rats in SGT group were injected with 100 micrograms of plasmid pcDNA3-gtfB, rats in control group with PBS solution. Genomes from submandibular gland, kidney, heart, liver, lung, and brain tissues were isolated later in 12 weeks. Genomes from different tissues were purified by low-melting agarose electrophoresis. Using the purified genomes as template, plasmid integration were examined by PCR(upper primer: 5' ATATGGTACCATGACCGAAGCGACATCTAAGCAAGA-3', lower primer: 5' ACTACTCGAGTTAGAACCATTGACCCTG AGCATTGC-3'). The sensitivity level of PCR was determined by adding gradient plasmid copies into genomes in control group. RESULTS: The examination of 6 tissues failed in revealing any evidence of integration at the sensitivity level that could detect 1 copy integration in 10,000 nuclei. CONCLUSION: The potential frequency of plasmid pcDNA3-gtfB integration into host cell genome would not exceed that of the spontaneous mutation. It was indicated that pcDNA3-gtfB was genetically safe as a promising anti-carious DNA vaccine. PMID- 12898773 TI - [The effect of telomerase inhibitors on oral squamous carcinoma cells]. AB - OBJECTIVE: To study the potential anti-tumor effect of telomerase inhibitors. METHODS: Human oral squamous carcinoma cell line KB was selected as target cell. The effects of 3'-azido-3'-deoxythymidine (AZT) and human antisense phosphorothioate (AS-ONS) for telomerase template on KB cell line were investigated. The cytotoxic effect of AZT and AS-ONS on tumor cells was quantified using MTT colorimetric assay. Assay of 3H-TdR incorporation was undertaken to measure the cell proliferation. The changes of telomerase activity after treatment was detected and quantified by telomeric repeat amplification protocol (TRAP) semi-quantitative analysis. The Flow Cytometry was used to detect apoptosis and measure cell cycle. RESULTS: Both AZT and AS-ONS inhibited the growth of KB cell line in a certain range of concentrations, and meanwhile the telomerase activity was reduced after treatment. In addition, both AZT and AS-ONS can induce apoptosis and arrest G1 phase of cell cycle. CONCLUSION: The results obtained above indicated that AZT and AS-ONS could be potentially used as an anti oral carcinoma agent or an auxiliary treatment for cancer. Those inhibitory effects might be partially due to the induction of apoptosis and the prolongation of cell cycle. PMID- 12898774 TI - [The study of effects of static magnetic field on SP-mRNA in trigeminal ganglion in rats]. AB - OBJECTIVE: To evaluate the effect of static magnetic field on the expression of SP-mRNA in TG in rats. METHODS: 44 Wistar rats aged 6-7 weeks were put into static magnetic field and were sacrificed at 1 h, 2 h, 6 h, 12 h, 24 h, respectively. In situ hybridization method was used to evaluate the changes of SP mRNA expression at different time point. RESULTS: Many neurons in TG were marked with SP probes in each group, the expression of SP-mRNA increased remarkably in static magnetic field group. In this group, the percentage of SP-mRNA positive neurons in TG increased greatly in 1 h, reached its peak in 2 h, from then on, decrease of the percentage started slowly but a moderate percentage was kept until 24 h, which was thought to be enough to maintain orthodontic tooth movement. The tendency of control group was almost the same with that of experimental group. The expression of SP-mRNA was higher in experimental group within 2 h but became lower after 2 h as compared with control group, this indicated that magnetic field reduced the SP-mRNA expression and exerted restoring effect on trauma. There were significant differences between experimental groups and control group at different time points (P < 0.01). CONCLUSION: The expression of SP-mRNA in TG in rats increased significantly in static magnetic field. PMID- 12898775 TI - [A study on the mechanical properties and microstructure of Alumina-zirconia nano composite infiltrated ceramic]. AB - OBJECTIVE: To evaluate the mechanical properties and microstructure of a new dental full-ceramic material: alumina-zirconia nano-composite infiltrated ceramic. METHODS: The flexural strength was tested with three-point bending method and the fracture toughness with single edge notch bend (SENB) method. The composition of crystal phases in the infiltrated ceramic was analyzed by X-ray diffraction (XRD). The microstructure of the infiltrated ceramic was examined by scanning electronic microscope (SEM). RESULTS: The average three-point flexural strength of the material was (610.85 +/- 37.07) MPa and the average fracture toughness determined by SENB method was (6.51 +/- 1.38) MPa.m1/2. The main crystal phases in this composite ceramic were alpha-Al2O3 and TZP-ZrO2. CONCLUSION: Alumina-zirconia nano-composite infiltrated ceramic is a new infiltrated ceramic with favorable mechanical properties. It demonstrated a promising future for clinical application. PMID- 12898777 TI - [Is general surgery dead?]. PMID- 12898776 TI - [A study on the thermal expansion characteristics of Alumina-zirconia nano composite infiltrated ceramic]. AB - OBJECTIVE: To study the characteristics of thermal expansion of the alumina zirconia nano-composite infiltrated ceramic. METHODS: The thermal expansion coefficients of alumina-zirconia composite matrix, infiltrating glass and infiltrated ceramic were determined by various methods, respectively. RESULTS: The coefficients of thermal expansion of composite matrix with 10 wt%, 20 wt% and 30 wt% zirconia were 7.607, 7.690 and 8.111 microns/(m. degree C) respectively. The thermal expansion coefficients of AZ-8 and AZ-10 infiltration glass were 6.867 and 7.333 microns/(m. degree C), respectively. The thermal expansion coefficient of infiltrated ceramic was 8.413 microns/(m. degree C). CONCLUSION: The thermal expansion coefficients of the glass and composite matrix matched well. The thermal expansion coefficients of the alumina-zirconia nano-composite infiltrated ceramic and several common veneering porcelains matched well, too. PMID- 12898778 TI - [Patient survival analysis in surgery of bronchogenic carcinoma from 1986 to 1997]. AB - Surgery is the most effective treatment of lung cancer (LC) in early stage disease (I-IIIa) of non small cell lung carcinoma (NSCLC). Even in resectable patients there are differences in survival accordingly to some prognostic factors. In 12 years period (1986-1997) there were 363 resected patients, in 350 of them lung cancer was confirmed. This group was analysed according to morphologic types, stages, and sex of patients, co morbidity and the type subsequent adjuvant therapy in relation to survival. Sensitivity and specificity of staging methods were also evaluated as well as the impact of staging accuracy to survival. Epidermoid cancer was found in 193 patients (55%), adenocarcinoma in 124 (36%), large cell-anaplastic type in 29 patients (8%) and small cell lung cancer in 4 patients (1%). Early postoperative mortality is 4%. 5 year after surgical resection survive 26% (90 patients). Other results are given in the text. Surgery of LC remains optimal therapeutic method. Survival of patients after lung resection doesn't change significantly in last years, even that we can use much more better diagnostic methods now. The way for the future seems to be the preoperative chemotherapy and the biologic target therapy. We need more precious staging procedures and more effective adjuvant therapeutic schedules in early as well as more advanced stages. PMID- 12898779 TI - [Principles of post-resection brachytherapy of solid tumors using high dose rate (HDR) afterloading]. AB - Postresection interstitial brachytherapy is one of the modern methods of radiotherapy the aim of which is to administer a larger radiation dose without greater irradiation of the surrounding sound tissues. The administration of higher radiation doses leads in some solid tumours to better local control of the disease and makes it thus possible to use the organ and its function after preserving surgical operations. Post-resection interstitial brachytherapy belongs in the wider concept to intraoperative radiotherapy the advantage of which is direct control of the irradiated area during surgery. The method of introduction of the radiation source into the applicators in the tumour during surgery (afterloading technique) makes its possible to start radiotherapy after obtaining the definite histological result. Post-resection brachytherapy reduces markedly the total time of irradiation treatment. PMID- 12898780 TI - [Surgery of hemorrhoids using the Long method and its complications]. AB - BACKGROUND: The aim of this article is an assessment of new surgical procedure- stapled hemorroidectomy according to Longo. We do concentrate on surgical complications and possibilities of it's management. METHODS: Prospective, clinical follow up of patients in which stapled hemorrhoidectomy was performed during the period of 2 years (1st December 2000--30st November 2002). Observation concentrates on surgical complications of this method. All patients had a clinical check up 3 weeks and 3 months after surgery. In case of any problems treatment and follow up continues. RESULTS: Stapled hemorroidectomy was performed during the period of 2 years in 52 patients (100%). There was 11 patients (21.2%) with some of surgical complication. The most serious one was massive rectal bleeding after surgery, which has been observed in 4 patients (7.6%). Other surgical complications observed in our group were anal stenosis, local infection, acute anal fissure and retention of urine. CONCLUSION: Stapled hemorroidectomy is now one of feasible alternatives for surgical treatment of hemorrhoids. Serious surgical complications observed in our patients were bleeding from the stapled suture line and anal stenosis. The aim of this article is to refer possible surgical complications of this method, it's prevention and management. PMID- 12898781 TI - [Intensive care medicine and its place among the medical specialties]. AB - The cost of providing intensive care is enormous. This resource consumption is focused on a relatively small number of patients. Over the past 30 years, despite new technologies and escalating costs, mortality rates for major disease conditions treated in the intensive care units remain unchanged. The studies done on organizational aspects of intensive care medicine do provide a strong argument that full-time intensivists improve the organization of intensive care, reduce mortality and increase efficiency of resource use. There are two general models of intensive care organization: open and closed. In the open model, an intensivist, if available, may provide consultation in particular department as needed. For patients with complex illness or prolonged length of stay, open systems are fragmented and poorly co-ordinated. In contrast, the closed system involves transfer of the patient to the exclusive care of a team of intensivists upon admission to the intensive care unit. The closed system typically provides formal triage, and consistent control, and mandated care by the intensivist team. Significant reductions in mortality rates and improvements in resource efficiency are seen with use of intensivist teams, a closed intensive care unit model and systematic triage. PMID- 12898782 TI - [Initial experience in use of the Freka Trelumina catheter]. AB - The authors present a report on their initial experience with the use of a Freka Trelumina probe. It is a nasal triluminal special probe for duodenal or jejunal nutrition and concurrent decompression of the stomach in case of its impaired motility and impossible oral nutrition. They draw attention in particular to the use in conservative treatment of acute pancreatitis and inform on favourable experience with the use of the probe under X-ray control. PMID- 12898783 TI - [Ultrasonography as an auxiliary method in diagnosis of acute appendicitis]. AB - The diagnosis of acute appendicitis remains a difficult task for even the experienced surgeon. For this reason every supplementary examination affords help for the surgeon its diagnosis. The goal of our study was to evaluate the significance of ultrasonography (USG) in diagnosing right lower quadrant abdominal pain. 204 patients admitted for suspect acute appendicitis were evaluated retrospectively. The accuracy of USG was studied by comparing this finding with preoperative, histopathological findings respectively. In comparison leucocytosis was also studied. In our studied cohort, USG examination had a sensitivity of 88.4% and specificity of 93.0%. Leucocytosis for diagnosis gave a sensitivity of 88.6% and specificity of 60.0%. PMID- 12898784 TI - [Adenocarcinoma of the small intestine: a rare diagnosis--4 case reports]. AB - The authors submit four case-histories of rare primary adenocarcinoma of the small intestine. The poor symptomatology and equivocal results of examinations caused a long delay of the final solution. As compared with data in the literature the submitted cases can be considered typical. PMID- 12898785 TI - Dreamscapes: portrayals of rectangular spaces in Doris Lessing's Memoirs of a Survivor and in dreams. PMID- 12898786 TI - Poor Ebenezer: avarice as corruption of the erotic and search for a transformative object. PMID- 12898787 TI - The cure of souls in the therapeutic state. PMID- 12898788 TI - Disquieting phantasy: Klein-Freud controversies. PMID- 12898789 TI - The changing place of the dream in psychoanalytic history, Part II: other perspectives, sociocultural influences and the challenges of neuroscience. PMID- 12898790 TI - Serum levels of inhibin B in men of different age groups. AB - The decline of testosterone levels in aging men is well documented. However, it is unclear to what extent the Sertoli cell marker inhibin B changes during aging. Herein we report on the determination of serum levels of inhibin B, follicle stimulating hormone (FSH), luteinizing hormone (LH) and testosterone in 906 patients from 16 to 89 years of age, presenting to our department for different complaints. There was a weak but significant correlation of the levels of inhibin B with age (r = 0.064, p < 0.05). A significant negative correlation of FSH and inhibin B was documented in all age groups (r = -0.423, p < 0.01). Also, the LH levels increased significantly with low inhibin B levels (r = -0.289, p < 0.01). Testosterone levels showed no significant correlation with inhibin B. We conclude from our study that Sertoli cell function as documented by serum levels of inhibin B is stable throughout life. In addition, the ongoing correlation of FSH and inhibin levels also indicates no significant decline of Sertoli cell function in the aging male. PMID- 12898791 TI - Assessment of andropause awareness and erectile dysfunction among married men in Ile-Ife, Nigeria. AB - Andropause (also known as androgen decline in aging males) has implications for the reproductive health and quality of life of older males. Very few studies have, however, been reported among the Nigerian population on andropause-related issues. This study assesses the perspective and level of awareness of married men in Ile-Ife, South-west Nigeria, of andropause. We also assessed their experience of erectile dysfunction, using a questionnaire based on the review of the International Index of Erectile Dysfunction. The study involved 355 married men, aged between 30 and 70 years. Our result shows a high level of misconception about andropause among our respondents, with 38.9% indicating that it is a myth, and another 23.6% attributing it to various causes other than being a natural aging process. We recorded a prevalence of erectile dysfunction of 43.8% (8.0% severe dysfunction and 35.8% moderate dysfunction). The prevalence of erectile dysfunction increased significantly with age, varying from 38.5% for age 31-40 years to 63.9% for the older age group of 61-70 years. The trend in prevalence of erectile dysfunction with age was significant (p < 0.05). An odds ratio of 2.82 (95% confidence interval 1.19-6.76) was recorded for the prevalence of erectile dysfunction at age 61-70 years compared with age 31-40 years. Our findings indicate a need for health education about andropause in Nigeria, and increased attention to the reproductive health concerns of males, and the older population. PMID- 12898792 TI - Oral testosterone undecanoate (Andriol) supplement therapy improves the quality of life for men with testosterone deficiency. AB - In a single-blind, placebo-controlled study, the effects of a 3-month oral administration of 160 mg/day testosterone undecanoate (Andriol) on the quality of life of men with testosterone deficiency were evaluated. The subjects included ten men with primary hypogonadism and 29 with andropause with sexual dysfunction as the most common problem. The changes in subjective symptoms were evaluated by the PNUH QoL scoring system and the St. Louis University Questionnaire for androgen deficiency in aging males (ADAM). Digital rectal examination (DRE) was performed and serum testosterone, prostate-specific antigen (PSA) and liver profile were monitored. Testosterone undecanoate treatment (n = 33) significantly improved sexual dysfunction and symptom scores of metabolic, cardiopulmonary, musculoskeletal and gastrointestinal functions compared to baseline and to placebo (n = 6). ADAM score also significantly improved after 3 months of treatment. Serum testosterone was significantly increased compared to pretreatment levels only in the testosterone undecanoate group. In the placebo group, no significant changes compared to baseline were found for testosterone levels and QoL questionnaires. No abnormal findings were detected on DRE or laboratory findings in either group. Adverse events, such as gastrointestinal problems and fatigue, were mild and self-limiting. It is concluded that androgen supplement therapy with oral testosterone undecanoate (Andriol) restores the quality of life through improvement of general body functions in men with testosterone deficiency. PMID- 12898793 TI - Oral testosterone undecanoate reverses erectile dysfunction associated with diabetes mellitus in patients failing on sildenafil citrate therapy alone. AB - AIMS: To evaluate the cause of failure of sildenafil citrate (Viagra) to restore erections in patients with organic erectile dysfunction (ED) associated with type II diabetes mellitus (DM) and receiving oral antidiabetic drugs. METHODS: Diabetic ED patients (n = 120), aged 43-74 years, failing to respond at least three times to 100 mg Viagra were evaluated. After at least 2 weeks' treatment with oral testosterone undecanoate (Andriol), 100 mg Viagra was used before coitus. ED was assessed with the International Index of Erectile Function (IIEF). Serum total testosterone, prolactin, thyroid stimulating hormone, lipid profile and prostate-specific antigen (PSA) were determined by standard methods and prostate volume by digital rectal examination. Age-matched diabetic ED patients (n = 100) served as controls for baseline values. RESULTS: Viagra non-responders had, at baseline, significantly lower testosterone and more depressed libido than controls. Andriol restored testosterone to normal levels and increased libido. In 84/120 (70%) Viagra non-responders, combined therapy with Andriol induced satisfactory erections, a significant increase in IIEF scale (question (Q) 3 from 2.0 +/- 0.2 to 3.7 +/- 0.3, Q4 from 1.9 +/- 0.1 to 3.4 +/- 0.2, Q12 from 1.0 +/- 0.1 to 4.2 +/- 0.4) and increased sexual contacts from 0.5 to 3-4 per month. No adverse events were noted, and PSA levels remained below 4 ng/ml. CONCLUSION: Decreased testosterone levels in patients with ED and type II DM receiving oral antidiabetic against may be responsible for failure to respond to sildenafil citrate therapy. Combination with oral testosterone undecanoate restores sexual function in these patients. PMID- 12898794 TI - Oxidative stress and antioxidant defense system in healthy, elderly men: relationship to physical activity. AB - The aim of the present study was to compare selected markers of oxidative stress and the antioxidant system between groups of men aged 65 or more, differentiated by their level of physical activity. Thirty-one community-dwelling healthy, elderly men (mean age 73.4 +/- 6.1 years) with a good nutritional status participated in the study. The Caltrac accelerometer was used to estimate energy expenditure related to physical activity. Subjects were divided into two groups according to the criteria proposed by Bauman: less active (< 1600 kcal/week) and more active (> 1600 kcal/week). In all the men, plasma total antioxidant status (TAS), thiobarbituric acid reactive substances (TBARS) concentrations and hydrogen peroxide (H2O2) levels were determined. The reduced glutathione (GSH) content and glutathione peroxidase (GPx) activity in hemolyzed red blood cells were also measured. There were no significant differences in TAS, H2O2 concentrations and the activity of GPx between less and more active men. In the group of more active subjects, plasma levels of GSH were higher, while TBARS concentrations were lower compared with the group of less active men (p < 0.05). Negative correlation between GSH and TBARS levels was observed (r = -0.71; p < 0.01), and GSH concentrations positively correlated with level of physical activity (r = 0.63; p < 0.05). The results indicate that habitual physical activity can favorably affect antioxidant potential and prevent lipid peroxidation in healthy, elderly men. PMID- 12898795 TI - Harmonized French version of the Aging Males' Symptoms Scale. AB - Interest in clinical investigations about the health-related quality of life (HRQoL) of aging men has increased in recent years. The aim of this paper is to inform the scientific community about a harmonized French Aging Males' Symptoms (AMS) Scale. There were two slightly different French AMS Scales, which both underwent an up-to-date linguistic and cultural translation process, i.e. were valid to be applied in research. However, it was felt to be unfortunate that two versions of one language in one country existed. Therefore, an ad hoc committee of both translation teams were asked to develop a harmonized single French AMS Scale. The harmonization meeting developed a consensus item-by-item and the new French reference scale was agreed upon. It was agreed that only this scale should be published to avoid confusion among future users. The French AMS Scale published in this paper should be used for future research and necessary cultural/linguistic adaptations in the French-speaking world. PMID- 12898796 TI - The aging male in the Old Testament. AB - According to the first books of the Old Testament--namely the Pentateuch--the antediluvian and post-diluvian patriarchs lived for hundreds of years having sons and daughters until the end of their exceptionally long lives. These unbelievable statements can be explained only if the particular character of the biblical chronologies is considered. The continence of David and the behavior of Solomon and of some of the 'judges of the people' pose very interesting historical, cultural and gerontological problems. PMID- 12898797 TI - The strengths and frailties of women with cardiovascular disease. PMID- 12898798 TI - Gender differences in cardiovascular risk factors. AB - Coronary heart disease is less common in premenopausal women compared to age matched men. This difference disappears in the post-fertile years, and is presumably related to the reduced levels of female sex hormones, with subsequent metabolic and hemodynamic modifications. Ovarian exhaustion induces a more atherogenic lipid profile, which may partly explain the increased risk of cardiovascular disease observed in post-fertile women as compared to age-matched fertile women. After the menopause, the combination of aging and estrogen deficiency negatively affects glucose metabolism. Diabetes mellitus blunts the beneficial condition associated with the female gender; furthermore, it increases the incidence of myocardial infarction, claudication and stroke in women more than in men. Finally, the unfavorable effects of menopause on the coronary risk seem to be mediated partly by changes in clotting and fibrinolytic factors. PMID- 12898799 TI - Gender differences in the neurohumoral control of the cardiovascular system. AB - The cardiovascular system is regulated by many complex neurohumoral mechanisms which ensure the cardiac, cerebral and renal functions. The nervous control of the heart is mainly mediated by the vagal and sympathetic systems and by their interaction, known as the sympatho-vagal balance. An increased sympathetic tone is found in many abnormal situations, such as arterial hypertension, diabetes, chronic heart failure and myocardial infarction, and is associated with an increase in over-all mortality. The hormonal control of the cardiovascular system is mediated by various substances such as renin-angiotensin, catecholamines, insulin and estrogens, that are themselves correlated with the autonomic nervous system. In contrast to men, fertile women show a predominant vagal tone. Sex related differences in the neurohumoral control of the cardiovascular system have been demonstrated during physical effort and in the hemodynamic adaptation to orthostatism. They have been postulated to explain the lower mortality in women compared to men among hypertensive or chronic heart failure patients. Prospective studies are needed to better define the gender differences in the pathophysiological mechanisms underlying cardiovascular diseases, in order to refine prevention and therapy. PMID- 12898800 TI - Gender differences in the clinical presentation to the emergency department for chest pain. AB - Previous studies suggest that the presentation, treatment, and outcome of acute coronary syndromes differ between women and men. Women arriving to the emergency department for acute coronary syndromes are usually older, have a higher prevalence of diabetes and hypertension, show a longer delay from symptom onset and have a slightly higher in-hospital mortality than men. In our experience including 4843 emergency department admissions for chest pain, the time to presentation was longer and diagnostic mistakes more frequent in women than in men, whereas medical treatment and the rate of revascularization procedures were similar. The introduction of Chest Pain Units with pre-defined diagnostic and therapeutic procedures should facilitate the early identification of highrisk female patients, reduce inappropriate hospitalizations, and improve the diagnosis and outcome of coronary heart disease. PMID- 12898801 TI - Treatment of patients with concomitant coronary and carotid lesions. PMID- 12898802 TI - Remodeling and recovery following myocardial infarction. PMID- 12898803 TI - Coronary endothelial dysfunction after ischemia and reperfusion and its prevention by ischemic preconditioning. AB - In the coronary circulation, when reperfusion follows ischemia, endothelial dysfunction occurs. This is characterized by a reduced endothelial release of nitric oxide and by an increased release of reactive oxygen species and endothelin. The reduced availability of nitric oxide leads to the adhesion of neutrophils to the vascular endothelium, platelet aggregation and, with the contribution of endothelin, vasoconstriction, which are responsible for the "no reflow" phenomenon. Neutrophil adhesion is followed by the release of the superoxide anion from neutrophils and endothelial cells. Preconditioning limits the endothelial damage by ischemia-reperfusion. A relevant role is attributed to the increased endothelial release of nitric oxide, while that of adenosine is controversial. Another effect of preconditioning on the coronary vasculature is the acceleration of vasodilation in reactive hyperemia after a brief coronary occlusion. The acceleration is prevented if myocardial protection is achieved by means of the activation of the mitochondrial adenosine triphosphate sensitive potassium channels by diazoxide and persists when ischemic preconditioning is induced after blockade of the same channels by 5-hydroxydecanoate. PMID- 12898804 TI - Atrial flutter: arrhythmia circuit and basis for radiofrequency catheter ablation. AB - The term atrial flutter was introduced 90 years ago for an arrhythmia with a unique electrocardiographic pattern. The development of endocardial mapping techniques in the last decade allowed the detailed characterization of the tachycardia circuit and the identification of the cavotricuspid isthmus as its critical part. This review stresses the position of atrial flutter in the new classification of atrial tachycardias and focuses on its unique electrophysiological characteristics and different variants described in humans. Transcatheter radiofrequency ablation across the cavotricuspid isthmus constitutes a feasible and safe therapy, which prevents flutter recurrences during the long-term follow-up. This paper describes the different techniques that validate bidirectional isthmus block, which is an important endpoint for successful ablation. PMID- 12898805 TI - Coarctation repair with prosthetic material: surgical experience with aneurysm formation. AB - BACKGROUND: Late aneurysm formation is a common complication after repair of an aortic coarctation with prosthetic material; its incidence varies between 5 and 46%. We reviewed our experience with the management of this complication and propose a radical surgical treatment, which has proved to be free from severe complications; furthermore, we suggest the possibility of a new percutaneous management of this complication. METHODS: From September 1974 to November 2002, 195 patients underwent primary repair of an aortic coarctation with prosthetic material (Dacron, polytetrafluorethylene or heterologous pericardium), with patch aortoplasty as the most common technique. During the follow-up period, reoperation for aneurysm formation was required in 13 asymptomatic patients. The diagnosis was made at angiography in 3 patients and at magnetic resonance imaging in 10. The indication for reoperation was an isthmic-diaphragmatic aortic diameter ratio > 1.5. Aneurysmectomy and tube graft interposition was performed in 12 patients; femoro-femoral cardiopulmonary bypass with a period of deep hypothermic circulatory arrest was carried out in 7 cases while 5 patients were submitted to normothermic atrio-femoral bypass; 1 patient underwent endovascular prosthesis implantation. RESULTS: There were no in-hospital deaths. Three patients experienced postoperative complications: bleeding (n = 1), left phrenic nerve paresis (n = 1), and chylothorax (n = 1). The mean follow-up period was 51.8 +/- 46.2 months; all patients were asymptomatic without clinical or instrumental evidence of recurrence. CONCLUSIONS: Aneurysm formation after primary repair of an aortic coarctation using prosthetic material is a potentially worrisome late complication and lifelong surveillance of these patients by means of magnetic resonance is mandatory. Surgical management, when indicated, has proved to be a definitive treatment and free from major complications. In highly selected patients, interventional management by percutaneous techniques may provide promising results. PMID- 12898806 TI - Exercise blood pressure response, cardiac output and 24-hour ambulatory blood pressure monitoring in children after aortic coarctation repair. AB - BACKGROUND: The aim of this study was to assess blood pressure at rest, the response to exercise and the 24-hour ambulatory blood pressure monitoring (ABPM) profile in children operated for aortic coarctation. METHODS: Twenty children were operated upon for aortic coarctation. The patients' data were compared with those obtained from 19 healthy controls of the same age. Treadmill exercise testing was performed and cardiac output was determined using the acetylene rebreathing method and indexed for the body surface area; ABPM was performed only in the patients group. The main outcome measures were the time of exercise, systolic (SBP) and diastolic (DBP) blood pressure both at rest and at peak exercise, maximal heart rate, total peripheral vascular resistance at rest and at peak exercise, and the pulse pressure (PP = SBP-DBP) at rest, at peak exercise and at ABPM. The Mann-Whitney test (non-parametric) and linear regression analysis were used when appropriate. RESULTS: Patients compared with healthy controls showed significant differences in SBP and PP at rest, and in DBP, cardiac index, total peripheral vascular resistance and PP at peak exercise. In the patients group only, linear regression analysis showed a significant correlation between PP and cardiac output, both at rest and at peak exercise, and between the arm-leg gradient at rest and PP at ABPM. CONCLUSIONS: These findings suggest that blood pressure abnormalities could be due both to the altered baroceptor reflex control mechanism, resulting in cardiac output and total peripheral vascular resistance abnormalities, and to the progressive increase in resistance during exercise at the site of the repair, resulting in the higher PP, that may be related to a local loss of the natural aortic elasticity. PMID- 12898807 TI - Preclinical thalassemic cardiopathy: a study by acoustic densitometry. AB - BACKGROUND: The cardiac function in thalassemia major has never been studied at ultrasonic backscatter techniques. We assessed the utility of acoustic densitometry in thalassemic patients without clinical or echocardiographic signs of heart failure. METHODS: Three groups of subjects with comparable age, sex and body surface area were analyzed: 25 with beta-thalassemia major (group A), 14 with thalassemia intermedia (group B) and 10 healthy subjects (group C). All patients were asymptomatic and without conventional echocardiographic signs of systo-diastolic dysfunction. The left ventricular mass and volumes were echocardiographically evaluated. The ultrasonic myocardial integrated backscatter signal (IBS) was recorded and analyzed by means of acoustic densitometry in the parasternal long-axis view at the septum and posterior wall, both at the basal and intermediate levels. Both the average image intensity and the systo-diastolic variations of the IBS (cyclic variation index-CVIibs and peak-to-peak intensity PPI), respectively related to the structure and contractility of the myocardium, were calculated. The serum ferritin and liver iron concentrations were also measured, as markers of tissue iron storage. RESULTS: The CVIibs was significantly lower in groups A and B than in group C at basal (22.7 +/- 8.4 vs 22.1 +/- 7.8 vs 31.8 +/- 10.2%; p = 0.001) and intermediate septum (24.4 +/- 7.6 vs 25.3 +/- 8.1 vs 30 +/- 9.8%; p = 0.03) and at basal (25.9 +/- 7.6 vs 24.5 +/- 6.1 vs 31.1 +/- 10.6%; p = 0.02) and intermediate posterior wall (25.1 +/- 5.1 vs 24.3 +/- 6.2 vs 30.2 +/- 6.6%; p = 0.02). The PPI was also significantly lower in groups A and B than in group C. Both CVIibs and PPI were comparable in groups A and B. The average image intensity and left ventricular mass and volumes were not significantly different in the three groups. No correlation was found between the densitometric findings and markers of tissue iron storage. CONCLUSIONS: In asymptomatic patients with thalassemia major with normal conventional indexes of systo-diastolic cardiac function, acoustic densitometry may show a reduced cyclic variation of the IBS as a possible marker of initial myocardial contractile deficiency. On the contrary, neither structural alterations nor the extent of myocardial iron stores are detectable by this technique in this type of patients. PMID- 12898808 TI - Iatrogenic aortic dissection during coronary intervention. AB - Iatrogenic acute dissection of the ascending aorta following coronary angiography and percutaneous intervention is rare. Localized aortic dissections have been treated by sealing the entry with a coronary stent. Extensive dissections may require a surgical intervention. We describe a coronary dissection with retrograde extension to the ascending aorta that occurred during angioplasty of the right coronary artery. The extensive dissection, which was limited to the ascending aorta, was successfully treated by stenting of the right coronary artery and monitoring the aortic dissection by means of transesophageal echocardiography. PMID- 12898809 TI - Postmenopausal hormone replacement therapy. PMID- 12898810 TI - Perspectives. Medicare overhaul: it's not just about drugs. PMID- 12898811 TI - Non-tumoral non-infectious intracranial calcifications. AB - Intracranial calcifications are often an accidental finding on conventional radiograms or computed tomography (CT) scans. They can be physiologic or pathologic, accompanying various diseases of the central nervous system. Pathologic calcifications can be found in tumors as well as in other non neoplastic diseases. In differential diagnosis of intracranial calcifications there should be mentioned two groups: infectious and non-infectious calcifications. In the paper there were discussed non-tumoral non-infectious intracranial calcifications. Although more sporadic than tumoral or infectious calcifications, calcified lesions in non-infectious diseases should also be taken into account in differential diagnosis of intracranial calcifications. PMID- 12898812 TI - Prevalence of infections of the Salmonella strains in the Lublin voivodeship and in Poland in the years 1980-2000. AB - The aim of the paper is presentation of the prevalence of infections of the Salmonella strains in the Lublin voivodeship in the years 1980-2000 in comparison with their nationwide prevalence in the same period. The analysed material comprised epidemiological data concerning the incidence rates of salmonelloses registered in Poland in the studied period. Since the beginning of the 1980s there was observed a constantly increasing trend in incidence rates of salmonelloses. After 1988 throughout Poland and after 1990 in the Lublin voivodeship there was noted gradual decline in incidence of infections caused by Salmonellas. Administrative decisions and recent improvements in sanitary situation in our country caused appearance of that favourable, decreasing trend of incidence rates of salmonelloses. The statistics concerning extraintestinal infections with Salmonellas are still unreliable, especially in the Lublin voivodeship. PMID- 12898813 TI - Foodborne infections and intoxications in the Lublin voivodeship in the years 1980-2000 in comparison with their prevalence in the Polish population. AB - The aim of the paper is presentation of foodborne infections and intoxications registered in the Lublin voivodeship in comparison with their nationwide prevalence in the whole country in the years 1980-2000. The analysed material comprised incidence rates of foodborne infections and intoxications, of which those caused by Salmonellas of animal source, registered in Poland and in the Lublin voivodeship in the studied years. In the analysed period in the Lublin voivodeship and throughout Poland there occurred a huge increase in incidence of foodborne infections and intoxications. The epidemics caused by Salmonella strains to the greatest extent influenced the increase in total number of cases of the disease, caused by deterioration in sanitary situation of the country, hygiene of the society and decline in quality of food of animal origin. The epidemiological situation of the disease in the Lublin voievodship is worse than nationwide average. PMID- 12898814 TI - Angiotensin-converting enzyme gene insertion/deletion polymorphism in Polish patients with myocardial infarction. AB - Both environmental and genetic factors play an important role in the pathogenesis of coronary artery disease. In view of angiotensin-converting enzyme (ACE) role in physiology and pathology and benefits of ACE-inhibitors therapy, the gene coding this protein meets criteria for candidate gene for the susceptibility to cardiovascular disease including myocardial infarction. The goal of our research was to assess the association of ACE gene I/D polymorphism with myocardial infarction in Polish population regarding different variables related to the disease (smoking, concomitant hypertension, obesity, plasma cholesterol level and family history). To detect ACE I/D polymorphism method of Rigat with some modification was used. The study population consisted of 314 individuals of Polish origin. 178 of them suffered from acute myocardial infarction or have survived it. Another 136 people without apparent signs and symptoms of cardiovascular pathology or diabetes served as control subjects. No one from the control group reported positive family history. Both allele frequencies and genotype distribution did not differ substantially between study groups. They were similar to those reported in other Caucasian populations. Further analysis did not detect any important differences in allele and genotype frequencies between experimental and control subjects regarding body mass index, total plasma cholesterol level and concomitant arterial hypertension. Cigarette smoking status influenced allele frequencies and genotype distribution in the group of patients with premature myocardial infarction. DD genotype was observed more often among non-smoking patients who suffered from myocardial infarction before the age of 50. These differences had reached the level of statistical significance (p < 0.05). CONCLUSIONS: 1. Insertion/deletion polymorphism in ACE gene does not seem to be associated with myocardial infarction in the studied Polish population. 2. The obtained results suggest that DD genotype may be associated with smoking dependent risk of myocardial infarction. Further studies are needed to prove this hypothesis. PMID- 12898815 TI - Dynamic computed tomography assessment of local extent and resectability of esophageal carcinoma. AB - The aim of this study was to define the usefulness of dynamic computed tomography in the preoperative assessment of esophageal cancer. CT scanning was performed in 93 patients with proven carcinoma of the esophagus. All results of CT were compared with surgical and pathologic findings. The analysis used the AJCC classification adapted to the needs of CT. The local extent of the primary tumour (T-stage) and its resectability were assessed and compared for different locations of the tumour. The overall accuracy of CT in tumour staging was 68.8%, 20.4% cases were overstaged and 10.4% were understaged. The highest sensitivity of CT has been obtained in estimation of tumours involving adjacent organs--T4- (86.5%) and the lowest for T3 (37.5%). Infiltration of surrounding vital structures could be proven with the positive predictive value of 69.7% and the negative predictive value of 97.3%. The efficiency of CT estimation for local tumour extent did not differ according to the tumour location. A high agreement (81.7%) was noted between CT results and intraoperative findings in assessing resectability. PMID- 12898816 TI - Dynamic CT of hepatic cirrhosis. AB - The aim of this research was the estimation of the dynamic sequential CT scanning (d-CT) method of the cirrhotic liver. The examined group consisted of 23 patients (8 women and 15 men, average age--57) histopathologically diagnosed as the cirrhotic liver. Dynamic CT scanning was performed 30-40 seconds after contrast material injection administered with an automatic syringe in the amount of 125 ml at flow time 2-3 ml/s. After the administration of contrast material the measurements of the hepatic parenchyma density were taken both in the left and right lobes. Time-density curves of the liver, spleen and portal vein were drawn for each patient. The curves of the liver and spleen of cirrhotic liver patients were not different from the curves obtained due to examining people with no medical problems. The time of peak enhancement of the liver and spleen of cirrhotic liver patients was delayed compared with control group. Time-density curves of the liver and spleen were considerably flattened of peak and were slow and longer in washout phase. Moreover, the time of peak enhancement of the portal vein curves was longer and the curves were flattened. Three cirrhotic liver patients had focal neoplastic lesion. The patterns of the time-density curves of the liver in dynamic CT scanning may contain diagnostic information useful for prophylactic monitoring of the neoplastic lesion, which develops in hepatic cirrhosis. PMID- 12898817 TI - Effect of nursing care on self-care efficiency of patients with hemiparesis. AB - The aim of the study was to determine the role of a nurse in rehabilitation of patients with hemiparesis. The study covered 60 patients--21 males and 39 females, aged 39-65, who underwent cerebral stroke and retained good verbal contact. While the patients were performing various instrumental-manual activities they were trained for self-care activities, and the effects were analysed. Self-care activities included hygienic activities and control of the body functions, such as: control of cardiovascular parameters, consumption of meals, control of sphincter muscles, prevention of muscular contractures and atrophy, as well as bed-sores and hemi-neglect syndrome. At Stage 1 of the study a considerable percentage of patients (approx. 50%) represented a lower category of self-care, while at Stage 2 the greatest number of patients attained a higher category (approx. 73.0%). In general, a considerable increase was noted in the percentage of patients who were efficient with respect to self-care, i.e. by 23.0%. PMID- 12898818 TI - Personality features of children treated due to vocal nodules. AB - There were estimated 14 features of personality of the children treated for vocal nodules. The purpose of study was definition of the selected features, which are typical of the examined group. The conclusion drawn from the conducted studies was: the children cured of the vocal nodules are more excitable, nervous, independent and often--individualists. They are inclined to leading and dominating. The results of the study suggest that psychotherapy can be completion of traditional treatment for vocal nodules in children. PMID- 12898819 TI - TEOAE after treatment of otitis media with effusion. AB - Otoacoustic emission is a noninvasive method of testing inner ear function. OAE is susceptible to the influence of many different factors of which the principal ones are the changes taking place in the middle ear. In transiently evoked otoacoustic emission (TEOAE) we deal with double signal transmission through the middle ear. This means that the sound reaching the cochlea depends on the function of middle ear. In the case of secretion into the tympanic cavity, suppression of the high-frequency otoemission components results from the effect of the mass increasing in the middle ear. The purpose of the study was the evaluation of otoemission in children treated with adenotomy and myringotomy due to MEE. The OAE value was estimated during the first 24 hours post surgery. The results of OAE study were compared with the clinical data, the level of hearing loss and the type of middle ear secretion. The study confirmed the presence of OAE in the majority of the cases. The results of the study indicate the applied treatment was effective. PMID- 12898820 TI - Plasma activity of interleukin-6 and some acute phase proteins in severe drug induced skin adverse reactions. AB - Plasma concentrations of Interleukin-6 (IL-6), C-reactive protein (CRP) and alpha 2 macroglobulin (alpha-2 MG) were examined in 9 patients with severe drug-induced cutaneous reactions (3 patients with toxic epidermal necrolysis and 6 patients with Stevens-Johnson syndrome). The activity of selected protein was measured using the immunoenzymatic ELISA method: a) in the acute stage of disease, before the treatment was applied and b) after clearing of skin lesions due to effective treatment. In the acute stage of the disease considerably increased plasma levels of IL-6 (p < 0.05) and both acute phase proteins (p < 0.001) were found. After clearing of clinical symptoms the concentrations of the examined proteins lowered towards control. But despite deep decrease, CRP and alpha-2 MG levels were still highly significantly increased (p < 0.001) in comparison with the control values. The results of this study indicate that in severe drug-induced skin reactions the acute phase response can be initiated and that the increased activity of examined proteins is longer observed than clinical symptoms of the disease. PMID- 12898821 TI - The frequency of different plural renal arteries rare variants. AB - The aim of this work was to evaluate the frequency of different plural renal arteries rare variants in healthy patients and in many different pathological diseases. Abdominal aortography with following selective angiography was performed on 35 randomly selected patients of Medical Department, with the use of radiopaque and with followed radiological parameters: P--75 kV, I--9 mA, T--10 sec. According to the available literature the following classification was introduced: double renal arteries--two vessels originating from the aorta, similar in diameter, blood supply areas, and their branches entering kidney through hilum (observed in 20% of cases), triple renal arteries--three vessels originating from the aorta with different diameter and blood supply areas, and their branches entering kidney through hilum (observed in 3%), upper pole accessory artery--originates from aorta, diameter of which being comparable to segmental arteries, entering kidney through hilum (6%), lower pole accessory artery (40%), upper pole perforating artery--originates from aorta or one of its major branches, diameter of which being comparable to segmental or subsegmental arteries, supplying one segment only, entering kidney outside of hilum (26%), and lower pole perforating artery (3%). PMID- 12898822 TI - Shift of zinc in serum and tissues in course of experimental acute pancreatitis. AB - The experiment was conducted upon 200 male rats of Wistar breed, weight 250-350 g, according to Heinkel and Aho's method. Four groups of rats were separated: Z- healthy (20), K1--control (60), K2--control (60), D--experimental (60). After 6, 12, 24, 48 hours since the acute pancreatitis was induced, the rats were anesthetized again (15 after each period), thoracotomy was performed and the blood from the left chamber was taken. Then after it turned into a clot, it was centrifuged for 20 minutes at 2000 rot./min. The blood serum obtained was then frozen in -25 degrees C until it was analyzed. The whole heart and right lung were taken out from the chest. The peritoneal cavity was re-opened and the pancreas, liver and left kidney were taken out. The method of the spectrophotometry of atom absorption was applied to mark the concentration of the chosen elements in the collected material. The SP 192 Pye Unicam spectrophotometer was used. The collected data were then statistically analyzed in order to define the arithmetic average and the standard deviation in the control group as well as in the examined groups. The statistic significance was defined by the application of the t-Student test and C. Cochren-Cox's test; accepting the value p < 0.05 for which the differences were taken as statistically significant. When marking zinc concentrations in the course of acute pancreatitis, the shift of this element in tissues was observed. The fall of zinc in blood serum, pancreas, kidneys, heart and lung was observed. The zinc increase was present in the liver only. Statistically significant differences were observed in the pancreas and liver in D and K2 groups, and in blood serum and kidneys in D group at 6th and 12th hour of the experiment. Differences observed in the heart muscle and lungs were not statistically significant. PMID- 12898823 TI - Odontogenic keratocyst in Gorlin-Goltz syndrome. AB - Gorlin-Goltz syndrome is an autosomal dominant disorder characterized by exteremly pleomorphic clinical manifestations including multiple odontogenic keratocysts. These cysts constitute only 3% to 11% of all jaw cysts, but are often of great concern to clinicians because of high recurrence rate, aggressive behavior and association with Gorlin-Goltz syndrome as they are found in 75-80% of the affected patients. In the paper there were presented histological, clinical, and radiological features of odontogenic keratocyst in Gorlin-Goltz syndrome. PMID- 12898824 TI - Styles of coping with stress among patients with allergic diseases. AB - Each man has his own characteristic and individual mode of coping in stress circumstances. The mode constitutes a permanent, personal inclination to a certain form of struggling with difficult situations. A mode of coping is at the same time a set of strategies applied in real confrontation with stress. The objective of the study is an attempt at analyzing the sources and the level of intensity of stress among patients with allergic diseases. The large majority, as many as 88.5% of the respondents claimed that their ability to cope with experienced stress was low or very low. Only 10.6% of the total number of the patients answered that they could cope with stress in this situation. PMID- 12898825 TI - The analysis of the structure of suicidal behaviours in the region of the Lublin voivodship in the years 1997-1999. AB - Suicide is a phenomenon that raises particularly strong emotions of the public creating a question about the sense of life and death. The phenomenon reflects customs, personal and social approach, as well as it undergoes historical and cultural changes. The aim of the study is to find the factors that influence making the decision about suicide. The analysis was concerned with the statistical data taken from the Voivodship Police Headquarters in Lublin and with death certificates in the Lublin voivodship. The highest suicide rates were for people included in the age group between 31 and 50. The suicides more frequently were males. PMID- 12898826 TI - Social support for patients with allergic diseases. AB - In recent years special attention has been paid to the issue of social support. So far there has been no special, applied, definition that would explicitly describe what social support is indeed. It results from the fact that the issue of social support has been of interest for numerous disciplines of science that have own fields of research and practical application. These are, among others, psychology, sociology, pedagogy, medicine. The objective of the study is an attempt to analyze the level of social support by people with allergic diseases. The research instrument was a self-structured inquiry sheet consisting of 25 questions and socio-demographic details. The strongest support for the patients was offered by the closest family (84%), followed by friends (51%), further members of the family (28.8%), acquaintances (26%) and institutions (14.4%). PMID- 12898827 TI - The behaviours of the inhabitants of the Lublin region in the situations of sudden cardiac arrest as reflected in the opinions of the Ambulance Service doctors. AB - Diseases of blood circulation system constitute the major cause of sudden cardiac arrest and deaths in Poland. Approximately 70% of all the cases of cardiac arrest occurs outside hospitals, frequently at patient's home. In the case of applying immediate resuscitation by the event witness in the situation of sudden cardiac arrest the percentage of the victims' survival is estimated at approximately 43%. Each delay of the initiation of Basic (BLS) and afterwards Advanced Life Support (ALS) lowers the chance of survival. Thus, it is extremely significant to educate Polish society in the area of basic resuscitative actions, particularly due to the fact that laypeople are usually the first ones at the scene of an accident. The aim of the paper was to determine to what extent the inhabitants of the Lublin region are prepared to rescue victims of sudden accidents by means of cardiopulmonary resuscitation and proper response in the situation of the sudden cardiac arrest. As results from the executed survey the inhabitants of the Lublin region in the situation of sudden cardiac arrest mostly do nothing but call an ambulance. The witnesses of the sudden cardiac arrest more frequently apply only rescue breathing or only indirect heart massage rather than rescue breathing and indirect heart massage simultaneously. The competence for applying CPR among the inhabitants of the Lublin region is poor and the resuscitative actions are much more frequently taken up by men. Premedical aid trainings are considered insufficient to prepare witnesses of sudden cardiac arrest for applying first aid. They should be conducted by doctors, in properly prepared and equipped training centers. The majority of the Ambulance Service doctors in Lublin indicated the demand for reminder premedical aid trainings. PMID- 12898828 TI - The usefulness of computed tomography spatial imaging in diagnosing lung tumours. AB - Diagnostic value of spatial CT reconstruction was analysed on the material of 17 patients with histopathologically confirmed lung tumours. Spatial 3D CT reconstructions of lung tumours were found to be a precious supplement of conventional CT examination providing important information concerning localisation, volume, surface structure, mutual relations of tumour and vessels as well as the evaluation of pleural infiltration. PMID- 12898829 TI - Antimicrobial therapy of upper respiratory tract infections in children. AB - Respiratory tract infections still represent a serious medical problem, mainly in people with different dysfunctions of the immune system. That kind of infections frequently occurs in the children and in the elderly. The aim of the present work was to estimate paediatrician's preferences in antibiotics prescription and therapy effectiveness in cases of ambulatory bacterial upper respiratory tract infections in children. Paediatricians working in the Lublin area were asked to fill in an inquiry that estimated their preferences to the prescription of antibiotics in children. Additionally, the microbiological examinations were performed determining the treatment effectivity. The inquiry was filled by 47 persons. The most frequently prescribed antibiotic was cefuroxime (93.6%), and the macrolids were prescribed rarely. During infection state, the predominant strain isolated was S. aureus (70.5%) and S. pyogenes (14.7%). After treatment, there was a significant increase in C. albicans isolation (44.1%) and H. influenzae (20.5%). There was a significant fall in isolation of S. pyogenes and S. aureus. Antibiotic prescription without earlier determination of susceptibility to antimicrobial agents had an influence on the appearance of postantimicrobial candidiosis, establishment of carrier state and increasing resistance among bacteria. PMID- 12898830 TI - Labyrinthitis in chronic otitis media. AB - Labyrinthitis is the most frequent complication of chronic otitis media (COM). Retrospective analysis of 1,210 patients operated due to COM was carried out. Circumscribed labyrinthitis was found in 17.2% and deafness in 1.2% of patients. To evaluate the effect of labyrinthitis on bone conduction (BC) we measured BC threshold in a group of 140 patients with unilateral COM without labyrinthitis and in a group of 42 patients with unilateral COM with labyrinthitis. BC threshold of the healthy ear was subtracted from the BC threshold of the affected ear in the same patient to eliminate presbyacusis. There was no significant difference in bone conduction threshold between the patients with circumscribed labyrinthitis and patients with COM without labyrinthitis. The incidence of labyrinthitis was higher in patients with cholesteatoma than in patients without cholesteatoma. Management of labyrinthine fistula is presented. PMID- 12898831 TI - The application of ultrasound contrast, 3D imaging and tissue harmonic imaging in the differential diagnosis of lymph nodes enlargement in children. AB - The application of Power Doppler mode examination introduced the assessment of vascularisation and measurement of blood flow parameters in lymph node's vessels as a differentiating criterion of benign and malignant lymphadenopathy. However, those criteria appeared insufficient in evaluation of the malignancy grade of enlarged lymph nodes, especially in the cases of lymph nodes with invisible or scantly visible vascularisation in Power Doppler mode. Introducing contrast media in ultrasonographic examinations enabling intensification of the Doppler signal even by 20 dB creates the hope of increasing diagnostic efficacy of ultrasonography in evaluation of vascularisation in lymph nodes enlargement. The purpose of the study was to define the usefulness of ultrasonographic contrast media (Levovist), 3D presentation and harmonic imaging in differential diagnosis of lymph nodes enlargement in children. 32 children with cervical lymph nodes enlargement underwent examination with ultrasonography. In the examinations, Levovist by Schering was used in concentration 300, the amount depending on patient's body mass. The analysis of results obtained in the study revealed that application of contrast media enables better visualisation of lymph node vascularisation. Localisation of the vessels which were not shown in conventional Doppler mode enables visualisation of the vessel architecture in the lymph node and better defining of vascularisation pattern. Application of the new methods of THI and 3D imaging and contrast media in Power Doppler examinations increases the diagnostic efficacy of ultrasonography in differentiating lymph nodes alternations. PMID- 12898832 TI - Taste perception of cigarette smokers. AB - In this work we have ventured to establish the dependencies between taste sensitivity for four basic tastes: sweet, salty, sour and bitter and endogenous factors such as sex and age and one exogenous factor, namely cigarette smoking. 471 randomly selected people, including 194 men and 277 women of different age were examined. Individual taste sensitivity was determined in the examined group, by means of saccharose, sodium chloride, acetic acid and quinine solutions. This examination showed that individual sensitivity limits range greatly to encompass varying concentrations of given taste substances. Values of individual sensitivity thresholds for stimulus substances and sensory minimum values obtained in this work showed that a substantial part of the examined people identified individual tastes at concentrations to a great extent exceeding mean threshold values. It can be supposed that those were cases of hypogeusia. There were also some instances of ageusia--a complete lack of sensitivity for sweet and bitter tastes. It was showed that cigarette smoking influences taste sensitivity of the examined people to a small extent. The changes observed were diversified and, in the majority of cases, statistically irrelevant. It can be assumed that taste receptor deficits caused by toxic compounds present in cigarette smoke are compensated by other senses. The conducted research pointed to no particular connection between identification thresholds of four basic senses and the age of the examined people. It can be caused by great diversification of individual threshold values. The research shows that, to a certain extent, taste sensitivity depends on gender. Generally speaking, women exhibit greater taste sensitivity in comparison to men. It seems that these differences are of hormonal nature. PMID- 12898833 TI - Concentration of mineral requirements in bone marrow tissue of patients with lymphoma malignum. AB - The concentration of microelements was tested in the bone marrow tissue of 20 patients with lymphoma malignum. The patients were not treated. There was no neoplastic infiltration in their bone marrows. The concentrations of magnesium, manganese, zinc, copper and ferrum were determined by the spectrophotometric method using atomic absorption spectrophotometer (Pye Unicam SP-192). The level of elements was compared to the value of blood morphology. The small value of MCV (mean corpuscular volume) was accompanied by a low level of copper, zinc and manganese although the values of MCH (mean corpuscular hemoglobin) and MCHC (mean corpuscular hemoglobin concentration) were normal. PMID- 12898834 TI - The influence of oxidative stress on the level of malondialdehyde (MDA) in different areas of the rabbit brain. AB - Lipid peroxidation is a process which is determined by the extent of peroxide forming free radicals mechanism and the peroxide-removing antioxidant system. To assess the peroxidation process, products of lipid peroxidation are analysed. Products of lipid peroxidation, such as thiobarbituric acid (TBA) are defined as thiobarbituric acid reactive material. In the present study we measured lipid peroxidation in three areas of the rabbit brain: brain hemispheres, brainstem and cerebellum. Supernatant and 10% homogenate were used to determine the level of malondialdehyde (MDA) according to the Ohkawa method. We measured levels of MDA before and after oxidative stress. Our results showed an important role of oxidative stress, because levels of MDA were increased significantly in comparison to the level of control, in all different areas of the rabbit brain. PMID- 12898835 TI - The eye lens evaluation of the atorvastatin-treated white rat. AB - The aim of this study is to determine potential cataractogenic activity of atorvastatin the 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitor. HMG-CoA reductase inhibitors (atorvastatin, lovastatin, simvastatin, pravastatin, fluvastatin, cerivastatin) (statins) are the most potent cholesterol and LDL-C lowering drugs. Statins differ in many aspects e.g. intensity of side effects. It is possible that some hydroxymethylglutaryl coenzyme A reductase inhibitors (statins) therapy is associated with cataract occurrence. The purpose of these studies was to focus on the potential cataractogenic contribution of atorvastatin. The studies were carried out on white Wistar rats. The animals were given atorvastatin (Sortis--Parke-Davis, USA) in two doses: 1.14 mg/kg mg/day, and 11.4 mg/day. Lens structure was observed in stereoscopic, dark-filed and in optic microscope, the cataract was observed in the examined preparations, specially high doses administered. We concluded that lens turned out to be reacting to atorvastatin. Drug dose corresponded to increase in the number and duration of cataract episodes (changes were more significant in the experimental groups 11.4 mg/kg). Alterations in the examined lens design may be a result of atorvastatin effect. PMID- 12898836 TI - The epidermis mitochondria ultrastructure evaluation of the etretinate-treated white rat. AB - Etretinate belongs to the group of drugs known as retinoids which are vitamin A derivates. It is a representative of second-generation retinoids, i.e. a synthetic, aromatic analog of transretinoic acid. The administration of etretinate induces clinical effects and morphological alterations in the structure of both pathological and healthy epidermis in men and animals. The aim of the present work was to examine the influence of long-term various doses of etretinate (Tigason) treatment on the ultrastructural design of white Wistar rats epidermis mitochondria. The mitochondria turned out to be reacting to Tigason. The drug dose-dependent changes as well as the ultrastructural mitochondria design, and the degree of other ultrastructural reproductive epidermis layer alterations cell organelle were observed. The increase in the mitochondria number and size was noticed. We concluded that etretinate application modulates the ultrastructural status of the epithelial mitochondria, and the administered drug dose correlates with the mitochondria ultrastructural design changes and the degree of ultrastructural alterations in the other organelle of the epidermis reproductive-layer cells. PMID- 12898837 TI - Information needs of the patients undergoing bone marrow transplantation. AB - The number of allogenic and autologous bone marrow transplantation (BMT) and the number of centers which perform these procedures have increased dramatically over the last 10 years. This increase can be attributed to the progress in supportive care which caused a significant decrease of transplant related mortality. Hematopoietic stem cell transplantation (HSCT) is now established therapy for many hematological malignancies, solid tumors and autoimmune disease. In spite of the great progress made, BMT is still a drastic procedure that may make patients suffer very much and can be even fatal. Careful exploration of the social and psychological situation of the patient is a very important factor which may prevent future difficulties. Many patients suffer psychological difficulties related to bone marrow transplantation from inadequate psycho-social preparation and support. It is important to prepare an information document about the newly diagnosed hematological malignancies. It must be a clear language and must answer the initial questions the patient will ask after diagnosis is made. The information leaflet distributed to the patients suffering from hematological malignancies can play an important role in the recovery of the immune system, thereby promoting a return to health. PMID- 12898838 TI - Palliative care--transiting old tradition and values into the modern health care practice. AB - Care of the dying patient is care for the patient who is still living, it is helping that patient to live his or her life to the fullest whether at home or hospice, or in hospital. It is care that is not just centered on the individual but includes members of the family. It is focused on improving the quality of patients' life, by incorporating into the professional care most of unique human values, such as respect, empathy, compassion and tender loving care. Palliative care is a broad band of care of indeterminate length, and does not necessarily end with the death of the patient. Until life begins to glow again, some surviving relatives may need support during the bereavement period. Thanks to Dr Elisabeth Kubler-Ross and Dame Cicely Saunders, two great women visionaries and leaders of the emergence of Palliative Care, one of the best things that has happened to medicine in the 20th century, the development of care for dying patients and their families was possible. PMID- 12898839 TI - Quality of care. Understanding the theoretical framework and learning the practical setting. AB - Different professional reports set out recommendations for the delivery of high quality services in cancer centers, units and primary health care. The explicit recommendations of the reports emphasized the need for cancer service to take account of the views and preferences of patients as well as of their families and careers. The assessment of the compliance with those recommendations is a base for the professional audit, measurement of the actual quality of care given, as well as the accreditation of cancer centers and units. Therefore, ensuring high quality in all areas of health care system is of great importance. PMID- 12898840 TI - Serum zinc (Zn) level dynamics in blood serum of patients with acute viral hepatitis B and early recovery period. AB - The aim of the present study was an analysis of serum zinc level dynamics in patients with acute hepatitis B and early recovery period compared with control group. The investigation included 39 patients aged 18-76 hospitalised in the Department of Infectious Diseases of the Medical University of Lublin, because of acute hepatitis B. Determinations of zinc (Zn) level in blood serum were made four times during hospitalisation and once, four weeks after discharging from the clinic in the early recovery period using atomic absorption spectrometry (AAS). The control group included 24 persons aged 22-69. Zinc (Zn) levels of those people were determined once. The obtained numerical data were subjected to statistical analysis. Lack of significant differences between men and women allowed to calculate the range of our norm, which was assumed at the level of M +/- 2SD, that is between 12.948-19.036 mumol/l. The significantly decreased serum zinc level was observed during hospitalisation while the differences stated in the serum level of this element in initial and early recovery determination compared with control group results are markedly at random. PMID- 12898841 TI - Prolactin concentration in the serum of male patients with chronic hepatitis C. AB - In this work we analysed the dynamics of prolactin serum concentration in male patients with chronic hepatitis C. A group of 52 men were included in the study, 26 of them constituted the control group. The diagnosis of the disease was confirmed by the presence of HCV-RNA in the serum and by the histological examination of the liver. None of the examined men was diagnosed with any co existing disease nor any infection markers of HAV and HBV were found. The prolactin concentration was determined in the serum of patients twice: on the 2nd day of hospitalisation and after 4 weeks of hospitalisation by the use of radioimmunological method (RIA) applying a prepared set of reagents RIA-PROL-CTK 4 (Sorin Biomedica, Italy). The results were statistically analysed. An analysis of the dynamics of prolactin serum concentration in male patients with chronic hepatitis C shows a statistically important increase in this hormone serum concentration. PMID- 12898842 TI - Fertility following laparoscopic uterine myomectomy in an infertile patient treated for 10 years. Case report. AB - The presence of myomas in the uterus is a relevant factor in infertility cases. Myomas may reduce contractility of the uterus, hinder migration of sperm and vascular changes within myomas themselves interfere with embryo implantation. The paper presents the case of a patient with over 10-year-history of infertility treatment diagnosed with numerous myomas of the uterus. The medical records revealed that the patient had undergone laparoscopy and laparotomy due to the left ovarian cyst and had been subjected to hysteroscopic removal of the uterine partition twice. The patient took part in the IVF programme twice--without success. At our Department the patient underwent laparoscopic myomectomy during which 5 myomas were removed: 4 subserous ones, 1.5-2 cm in diameter and an intramural one, 6 cm in diameter. The sites of the biggest myoma were laparoscopically sewn putting in 3 single sutures. In her next cycle the patient spontaneously conceited. The pregnancy was without serious complications and at 38 weeks was terminated by Caesarian section. The infant was delivered in good general condition (10 Apgar points) and weighted 3,360 g. On Caesarian section no uterine dehiscence following laparoscopic myomectomy was observed. PMID- 12898843 TI - Histological examination of the extrabulbar segment of the optic nerve in experimental animals after administration of Cladribine. AB - The experiment was carried out on rabbits, females of New Zealand breed, weighting about 3 kg. Rabbits from the experimental group I received Cladribine in the dose corresponding to the schema of the experimental treatment in the hairy cell leukemia and animals from the experimental group II, the dose corresponding to the experimental treatment in multiple sclerosis. The optic nerves were collected for histological examinations in the light microscope. It appeared that administration of Cladribine in the dose corresponding to the therapeutic dose used in therapy of hairy cell leukemia and sclerosis multiplex does not cause evident morphological changes in the extrabulbar segment of the optic nerve on the level of the light microscope. PMID- 12898844 TI - Influence of the new medicine--2-CDA (Cladribine) on the ultrastructure of the extrabulbar segment of the optic nerve in rabbits of New Zealand breed. AB - Experiments were carried out on the rabbits of New Zealand breed weighting about 3 kg. Rabbits from the experimental group received Cladribine in the dose of 0.07 mg/kg/24 h each morning subcutaneously for 6 days, three cycles with 5-week intervals. Specimens of the optic nerve were stained according to the Reynold's method and observed in Tesla BS-500 transmission electron microscope. Results achieved from examinations of slides in experimental group indicate that Cladribine administered in the dose corresponding to therapeutic dose used in humans for experimental treatment of sclerosis multiplex does not cause the damage of extrabulbar segment of optic nerve in experimental animals. The achieved results suggest necessity of new research works regarding the eye structures of mesenchymal origin. PMID- 12898845 TI - Protective effects of ACTH (4-9) in degeneration of hippocampal nerve cells caused by dexamethasone: ultrastructural studies. AB - The purpose of this research was an assessment of protective action of ACTH (4-9) in dexamethasone-induced neurodegeneneration on the base of ultrastructural examinations of hippocampal neurons in the CA3 region. The experiments were carried out on adult Albino Swiss mouse males. The animals were divided into three groups: control group, experimental group 1-dexamethasone 8 mg/kg/24 h for 28 days, experimental group 2-dexamethasone and ACTH (4-9) 50 micrograms/kg twice a week. Results of our investigations show that ACTH (4-9) prevents neurotoxic influence of dexamethasone and its protective action is connected with the ability to inhibit degenerative processes in neurons having a character of apoptosis. PMID- 12898846 TI - Immunocytochemical examinations of hippocampal nerve cells after experimental administration of dexamethasone. AB - The purpose of this research was an assessment of MAP2 immunoreactivity in hippocampal neurons after administration of toxic doses of dexamethasone. Experiments were led on Albino-Swiss mouse males. The obtained results indicate that dexamethasone causes significant decrease of MAP2 immunoreactivity in hippocampal nerve cells of the CA3 region. Our results show damage of neuronal cytoskeleton in this area of the brain. PMID- 12898847 TI - Neuroprotective effect of ACTH (4-9) in degeneration of hippocampal nerve cells caused by dexamethasone: morphological studies. AB - The purpose of this research was an assessment of neuroprotective effect of ACTH (4-9) in degenerative changes of nerve cells induced by dexamethasone. Experiments were led on Albino-Swiss mouse males. We examine morphological changes of neurons in the dorsal hippocampus in slides stained with cresyl violet and we performed quantitative analysis of neurodegenerative changes using a computer analyser of histological pictures. Achieved results indicate that ACTH (4-9) shows neuroprotective effect against neurotoxic influence of dexamethasone. This chemical inhibits dexamethasone induced degeneration of hippocampal nerve cells having morphological features characteristic of apoptosis. PMID- 12898848 TI - The influence of nitroglycerin on myocardial 99mTc-Tetrofosmin uptake defects in coronary artery disease. AB - Nitroglycerin (Ntg) is known to increase blood flow to the hypoperfused myocardial area and thus to reduce regional ischaemia of the heart muscle in coronary artery disease (CAD). The aim of this study was to assess the effect of a therapeutic dose of Ntg on 99mTc-Tetrofosmin uptake defects occurring in patients (pts) with CAD in myocardial SPECT at rest. The study population comprised 34 pts with CAD. All subjects underwent myocardial perfusion SPECT at rest, using 2-day, protocol. On the first day 99mTc-Tetrofosmin was injected in baseline conditions and on the second day, after sublingual administration of 0.5 mg Ntg. The baseline examination revealed altogether 78 Tetrofosmin uptake defects in all 34 pts. The SPECT after Ntg showed improved tracer uptake in 33/78 perfusion defects in 18/34 pts, no change in 29/78 defects in 7/34 cases and more reduced tracer uptake in 16/78 defects in 9/34 pts. No explicit, significant relationship was found between 99mTc-Tetrofosmin uptake changes after Ntg on the one hand and the number of involved vessels, degree of coronary artery stenosis, history of myocardial infarction and LV global and regional function on the other. It is concluded that sublinqual administration of Ntg may result in improvement as well as in deterioration of regional perfusion defects, assessed with 99mTc-Tetrofosmin in CAD. The relationships between Ntg-augmented changes in uptake defects of this tracer on the one hand and conditions of the LV function and severity of heart muscle perfusion abnormalities on the other are unclear and should be verified on a larger study population. PMID- 12898849 TI - Attitudes of young people towards problems connected with human procreation. AB - The aim of the paper was to get acquainted with opinions and attitudes of secondary school students concerning the issues of human procreation. The survey was carried out in secondary comprehensive and technical schools in Lublin during the years 1999 and 2000 among 300 students aged 16-19. The survey shows that mass media play the leading role in conveying knowledge and shaping attitudes and behaviours of teenagers in the sphere of sexuality. The function of family and school in this field is realized to an insignificant extent. Media make use of issues and interests of teenagers that are natural at certain age and succeed in promoting acceptance of early sexual relationships, not showing the risk connected with them at the same time. Among the group under the survey 37% of the respondents have already had sexual initiation. Girls accounted for 41.5% of the total and boys--for 58.5%. The average age of sexual initiation among the respondents was 15.87. The declaration of love, according to the majority, gave the right to take up sexual activity. They accepted the rule that you can make sex whenever you wish and a stable partner is not necessary, which is a sign of psychological immaturity and willingness to grow up fast. The use of contraception was approved of by nearly all the respondents. The most frequent preventive methods they used were: condoms (44.8%), withdrawal method (15.3%), both characterized by easy accessibility but at the same time low efficiency. 7.6% of sexually active students do not use any method of contraception. Such behaviours create a serious risk of premature pregnancy and sexually transmitted diseases. PMID- 12898850 TI - Hypoplastic left heart syndrome: morphological study of 21 autopsy cases. AB - Hypoplastic left heart syndrome is a group of relatively rare congenital malformations associated with severe obstruction of the left outflow tract. Twenty-one cases of Clinical with the syndrome were found in the autopsy files of the Department of Pathomorphology, Medical University of Lublin in the years 1991 2000. Most children were males, delivered at term, with normal body weight. None of them was surgically treated nor survived longer than one month. The most common subtype of valvular malformations included in the syndrome was aortic stenosis with mitral stenosis. In 15 cases microscopic examination revealed: recent myocardial necrosis, interstitial fibrosis and focal calcification. In 6 cases the syndrome coexisted with the congenital malformations of other systems. PMID- 12898851 TI - Asymptomatic bacteriuria in patients on continuous ambulatory peritoneal dialysis. AB - Chronic inflammatory state and chronic infections appear to be one of the most important risk factors of morbidity and mortality in patients (pts) on dialysis. The aim of our study was to determine the frequency of urinary tract infections (UTI) in clinically asymptomatic patients on CAPD. The study included 43 (22F, 21M) clinically stable asymptomatic CAPD patients with diuresis ranged from 300 to 2500 ml per 24 h. Clean catch urine specimen, was cultured and identified by standard technique. Urine from 31 patients had no significant organism growth, from 12 patients grew over 100,000 colonies per ml; 8 patients had been infected with E. coli, 2 pts with Proteus mirabilis, one with Enterococcus gallinarum, one with Klebsiella oxytocea. 7/12 infected pts were diabetic, 67% infected and 13% not infected pts had pyuria. CONCLUSIONS: Immune system disturbance and renal failure can predispose to common asymptomatic UTI in CAPD pts. E. coli is the most common bacterial strain found in infected dialysis pts. Diabetes mellitus is and additional predisposing factor for UTI in patients on CAPD. Pyuria is a valuable parameter of UTI in CAPD patients. PMID- 12898852 TI - Changes of surface tension of dialysate depending on its composition in continuous ambulatory peritoneal dialysis. AB - Surface phenomena resulting from interactions among molecules occur commonly in nature. Surfactants, substances with surface activity, have a big influence on surface tension. They reduce surface tension when added to the solution, even in minimal concentration. The behaviour of surface tension of the dialysate is difficult to properly assess, because of its complex composition. Dialysate is a mixture of organic and inorganic substances, both macro- and micromolecular. Among them are proteins, phospholipids, fatty acids. In the literature there are no publications showing the behaviour of surface tension of the dialysate. The aim of the study was the assessment of changes of dialysate surface tension in non-complicated continuous peritoneal dialysis (CAPD), depending on time of the presence of dialysis fluid in the peritoneal cavity and on the concentration of different substances in it. The study was performed on 39 dialysate samples obtained from 6 patients chronically treated with continuous ambulatory peritoneal dialysis (CAPD). During the study patients had no symptoms of peritonitis. Surface tension of the dialysate was determined, using the Wilhelmy's method. Ten measurements were performed in every sample of the dialysate and the mean values and standard deviation were calculated. Simultaneously the concentrations of sodium, glucose, urea, creatinine, total protein were measured in the dialysate. As a result of the study a significant negative correlation between dialysate protein concentration and surface tension was found. The significant negative correlation between surface tension and the dwell time was also found. There was no significant correlation between the value of surface tension of the dialysate and concentrations of glucose, urea, creatinine and sodium. PMID- 12898853 TI - Effect of acetate and bicarbonate dialysate on whole body bioimpedance (BIS) and segmental (thoracic) bioimpedance in hemodialysed (HD) patients. AB - There are sparse data about the influence of dialysis itself on bioimpedance measurement. The aim of this study was to evaluate the influence of dialysis without ultrafiltration on bioimpedance of thorax and bioimpedance of the whole body in hemodialysed patients. Isovolemic HD was performed during the first hour of dialysis session, using bicarbonate and then acetate dialysate. Thoracic impedance, whole body bioimpedance, blood pressure and blood density were measured. After bicarbonate dialysis significant TI lowering was observed with simultaneous R increase. There was a significant blood density lowering. Acetate hemodialysis caused no significant changes in TI but significant increase in R values. Blood density did not change after acetate HD. The influence of other factors, not only hydration status changes, must be considered in interpretation of bioimpedance measurement analysis. PMID- 12898854 TI - Adequacy of hemodialysis. AB - The concept adequate includes blood purification, maintenance of electrolyte fluid, and acid-base equilibrium. Dialysis can completely reverse the retention of some small molecules, electrolytes (K, Na, Mg), water and acid-base, and has no effect on anemia, hypertension, arteriosclerosis, cardiomyopathies, malnutrition, bleeding, or partial effect on Ca-P balances. Other therapeutic interventions are necessary to optimally treat the uremic syndromes. Urea kinetic modelling is probably the most objective way of measuring the efficiency of dialysis but we must remember that there are numerous alterations in ESRD which cannot be measured by urea kinetic modelling. PMID- 12898855 TI - Diagnostic value of HRCT and 3DCT in the assessment of chronic maxillary sinusitis. AB - In a group of 37 patients with chronic maxillary sinusitis in whom CT examination had been performed diagnostic value of secondary spatial reconstructions and high resolution sections was analysed. It was found that the supplementation of standard CT examination with high-resolution sections and spatial pictures increases diagnostic possibilities in the assessment of maxillary sinusitis as for recognising, differentiating and determining anatomical etiologic factors. PMID- 12898856 TI - Wegener's granulomatosis--exacerbation with pulmonary manifestation. AB - There was described a case of a 56-year-old woman with lesions in chest X-ray. Three years before she was treated in the Department of Laryngology of Medical University of Lublin because of ulcerations of nasopharynx. There was established diagnosis of Wegener's granulomatosis. PMID- 12898857 TI - Concentrations of branched amino acids: isoleucine, valine, leucine in serum of hemodialysis patients during one-year observation. AB - Malnutrition is commonly reported in haemodialysis patients (HD pts) and is multifactorial in origin. Previous studies indicated a possible relationship between plasma levels of branched-chain amino acids (BCAA): isoleucine, leucine, valine and nutritional status of HD pts. The BCAA have important influence on metabolism of energy in skeletal muscles, in addition the leucine makes easy the synthesis of protein. In the present investigation we evaluated some parameters of nutritional state and the concentration of branched-amino acids in haemodialysis patients during one-year observation. Sixty-seven patients undergoing haemodialysis were assessed in the first study (study I), after 12 months this assessment was repeated in 70 patients (study II). Blood was drawn before the midweek dialysis session. Morphology, plasma albumin concentration and urea for KT/V calculation were determined by routine methods. Branched amino acids were measured by chromatography method with amino acid analyser AAA 400, using a lithium-citric buffer system. The plasma amino acid pattern was measured in the study group and compared with 20 healthy persons. Mean amino acids concentration observed in both study groups was significantly lower than in the control group. Mean concentration of BCAA was also lower than in control group. There was a small negative correlation between albumin concentrations and the length of dialysis therapy. There were significant positive correlations between: total amino acids, branched amino acids as well as albumin, especially in study II. PMID- 12898858 TI - Genetic polymorphisms of renin-angiotensin system and progression of interstitial nephritis. AB - Genes of the renin-angiotensin system (RAS) are involved in the progression of renal failure. Among them, the angiotensin-converting enzyme (ACE), angiotensinogen (AGT) and angiotensin II type 1 receptor (AT1R) genes are of particular interest. We examined polymorphisms of these three genes for association with the development of interstitial nephritis and progression to end stage renal failure. The allele frequency and genotype distribution were compared in 90 patients with interstitial nephritis and 200 healthy controls. DNA samples were genotyped by polymerase chain reaction (PCR). We did not find statistically significant differences between groups in the insertion/deletion polymorphism of the ACE gene. An involvement of M235T polymorphism of the AGT gene in renal disease was observed in our study. The frequency of the T allele was higher in patients than in controls (32% vs. 24%). In the A1166C AT1R polymorphism the homozygous CC genotype was also more frequent in interstitial nephritis patients (7% vs. 3.5%). In patients carrying the C allele, an average time to ESRD was significantly shorter than in subjects with the AA genotype. Our study shows the association of the AGT and AT1R gene polymorphisms with the development and progression of interstitial nephritis. The C allele of the A1166C polymorphism appears to be a risk factor for faster disease progression. PMID- 12898859 TI - Changes of extracellular volumes measured by whole and segmental bioimpedance analysis during hemodialysis in end-stage renal disease (ESRD) patients. AB - The bioimpedance technique is postulated as not expensive and non-invasive method for measurement of body water compartments in humans. However, the accuracy of the whole body bioimpedance technique has been questioned by several authors because of nonhomogenic nature of the human body. In the present study we used the whole body bioimpedance technique in comparison to segmental measurement from the arm and leg for assessment of extracellular water compartment. The extracellular water compartment value as measured by the whole body technique significantly correlated with extracellular water compartment value measured from segmental measurement in the leg. We postulate practical use of segmental measurement from the leg as an indicator of body hydration. PMID- 12898860 TI - Nutrition and hydration status improve with exercise training using stationary cycling during hemodialysis (HD) in patients with end-stage renal disease (ESRD). AB - Patients with end-stage renal disease (ESRD) undergoing hemodialysis and peritoneal dialysis are functionally limited as a consequence of their physical, emotional, and social problems. Exercise intolerance is a major problem in chronic renal failure. Stationary cycle training during hemodialysis is recommended as safe, effective, and practical in ESRD patients treated on hemodialysis. The aim of the present study was to evaluate the effect of cycling exercises in 10 HD patients during 6-month period (including each of dialysis sessions) on nutrition, dialysis adequacy, and fluid parameters as measured by biochemical, and bioimpedance parameters. A significant increase in serum albumin concentration, Kt/V, and nPCR, and decrease in serum CRP have been observed after 6 months of regular stationary cycling during hemodialysis. Relative changes (pre post HD) in extracellular water compartment and ECW/TBW ratio have significantly increased after 6 months of observation period. PMID- 12898861 TI - How are hemodialysis patients treated in the Lublin region--based on the selected issues of diagnosis and therapy. AB - The aim of the paper was to answer the questions about the quality of life of dialysis patients treated in the selected dialysis centres of the Lublin region in relation to the recent quantitative and qualitative breakthrough in dialysis therapy in Poland. The studies including 288 patients of 7 medium-sized centres were performed using the questionnaires filled in by interviewers. The present paper describes the results of analysis of the questions addressed to the centre staff but concerning the way of treatment of a particular patient. The results show that to follow the world-wide trends, chronic dialysis treatment should be of an optimum quality and the methods evaluating it widely propagated. Moreover, much is to be done in the field of the latest diagnostic procedures and therapy of dialysis patients. On the other hand, the technical aspects of dialysis therapy look quite good. To evaluate the changes in the aforementioned processes more accurately, it should be worthy to broaden the studies including the remaining centres of our region and to continue them in the future. PMID- 12898862 TI - Care provided for hemodialysis patients in selected dialysis centres of the Lublin region. AB - The aim of the study was to answer the questions about the quality of dialysis for the patients treated in the selected dialysis centres of the Lublin region in relation to a quantitative and qualitative breakthrough observed in the field of Polish dialysis therapy in recent years. The study including 288 patients in 7 medium-sized centres used the questionnaire filled in by interviewers. The present paper analyses the patients' answers to the questions about medical care provided in a particular centre. It is difficult to interpret the results explicity since there are no available standards of such management. We believe that this study could be used to define them. Nevertheless, the authors think that medical care for the patients treated in dialysis centres of our region is good, although only the patients' answers were analysed. To evaluate possible changes in the processes mentioned above, it should be worthy to broaden the studies including the remaining centres of our region and to continue them in the future. PMID- 12898863 TI - New aspects of osteoporosis--a literature review. AB - On the basis of the literature the author describes some aspects of osteoporosis (pathogenesis, diagnosis, prophylaxis, and treatment). He focuses attention on the new factors which take part in this process such as: markers of bone turnover, local factors--cytokines and genetics. He also notices that osteoporosis can concern maxilla and mandible, therefore, maxillofacial surgeon should know this disease. PMID- 12898864 TI - Activity of superoxide dismutase and glutathione peroxidase in human semen in normozoospermia and spermatopathy. AB - The aim of the study was to assess the levels of antioxidative enzymes- superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in the whole semen of men treated for infertility. The first group consisted of the patients with normozoospermia; the second--included the men with oligozoospermia. According to sperm motility, the patients were divided into the third group--motility higher than 50% and the fourth group--motility lower than 50%. The study showed that the levels of both enzymes were higher in men with oligozoospermia than those with normozoospermia. It was also found that the activities of SOD and GSH-Px were higher in men with low motility(< 50%) compared to high motility patients. The results confirm that the activity of superoxide dismutase and glutathione peroxidase is related to the parameters of human semen assessed according to the WHO standards. PMID- 12898865 TI - Histological examination of the retinal ganglion layer in rabbits after experimental administration of the new immunosuppressive medicine--Cladribine. AB - The experiment was carried out on rabbit females of New Zealand breed weighing about 3 kg. Animals from the experimental group received Cladribine according to the schema of experimental treatment in the hairy cell leukemia and rabbits from the experimental group II--according to the schema of experimental treatment in multiple sclerosis. Specimens of retina were collected for histological examinations in the light microscope. It was revealed that administration of Cladribine in the dose corresponding to the therapeutic dose used in human for the treatment of the hairy cell leukemia and multiple sclerosis does not cause evident morphological changes in retinal ganglion cells on the level of light microscope. PMID- 12898866 TI - Ultrastructural changes of retinal ganglion cells in rabbits of New Zealand breed after experimental administration of 2-CDA (Cladribine). AB - The experiment was carried out on rabbit females of New Zealand breed weighing about 3 kg. The animals from the experimental group received. Cladribine in the dose of 0.1 mg/kg for 7 days. Specimens of retina were collected for ultrastructural examinations. It was revealed that administration of Cladribine in the dose corresponding to the therapeutic dose used in human for the treatment of the hairy cell leukemia causes morphological changes in ultrastructure of retinal ganglion cells which are classified as reversible in cytophysiology. The necessity of periodic, regular ophthalmological examinations during Cladribine treatment was indicated. PMID- 12898867 TI - Zinc and copper in the human placenta membranes. AB - This paper determines the distribution of bioelements such as zinc and copper in the membranes of the placenta, i.e. the chorion and the amniotic sac. The study material was taken in the course of labour from women with normal pregnancy. In the material taken, the level of microelements was determined by nuclear absorption spectrometry. In the chorion a higher level of zinc and copper was found in comparison with the amniotic sac. PMID- 12898868 TI - The activity of dental caries in students of the Faculty of Stomatology examined by using microbiological and biochemical tests--Dentocult LB. AB - The aim of the study was to examine the relation between the value of Dentocult LB and numbers of DMF, DMFs and Ds in the 20-25 age group. Sixty-six randomly chosen students of the Faculty of Stomatology, Medical University of Lublin were included in the study. In all of them, their dental state was assessed by using average numbers of DMF, DMFs and Ds. In the microbiological examinations, a ready made medium from the Dentocult LB set was used. Statistically significant, directly proportional dependence of the number of surfaces with Ds active caries and the LB number was found. PMID- 12898869 TI - Can nonsteroidal anti-inflammatory drugs favourably affect the alimentary canal? AB - Nonsteroidal anti-inflammatory drugs (NSAIDs) represent one of the most commonly used therapeutic drug groups world wide. 1.5% of the world's population is estimated as taking NSAIDs. Although NSAIDs are still generally believed to have toxic effects on the alimentary tract, some evidence of their favourable effects on the alimentary canal has recently been reported. Wider knowledge concerning these effects may allow us in the future to use NSAIDs as preventive measures in carcinoma or adenocarcinoma of the colon or even in carcinoma of the esophagus, in the treatment of motor disorders of the alimentary tract or inflammatory diseases, e.g. reflux esophagitis. The clinical studies evaluating the NSAID effects on the treatment of acute biliary colic and the short-term administration of small doses of aspirin effective in secondary prevention of cholelithiasis are also encouraging. At present the studies are being carried out to show the therapeutic action of NSAIDs in pancreatitis. PMID- 12898870 TI - Disadvantageous effects of nonsteroidal anti-inflammatory drugs on the alimentary tract. AB - Nonsteroidal anti-inflammatory drugs (NSAIDs) are the most frequently recommended pharmaceutical agents worldwide. These drugs are known to damage the gastric and duodenal mucosa and also esophagus, small and large intestine mucosa. Severe complications are thought to develop in 1-2% patients after the 3-month treatment with NSAIDs. The most severe complications include erosions, ulceration accompanied by bleeding and perforation. These severe adverse reactions may occur in patients without any complaints. The factors of increased risk of complications during NSAIDs therapy are well known. The most important are: morbus ulcerosus disease, above 60 years of age, larger doses of NSAIDs, simultaneous steroid therapy, anticoagulants and Helicobacter pylori infection. NSAIDs may cause strictures and enteropathy in small intestine; and may exacerbate already existing diseases of the large bowel or also cause strictures. So, before NSAIDs prescribing one should take into account all possible harmful effects and identify the patients who are particularly at risk. PMID- 12898871 TI - Histological and ultrastructural investigations of placenta villi in intrauterine growth restriction of fetuses (IUGR). AB - The investigations were carried out on five placentas derived from normal pregnancies and on the five placentas derived from the pregnancies with intrauterine growth restriction (IUGR) of fetuses. Semithin sections were stained with methylene blue and azure and estimated by light microscope. Ultrathin sections were stained according to the Reynold's method and estimated by the electron microscope. Two types of irregularity were observed: 1) lack of vessels with a concomitant increase of macrophage number in the stroma of villi, 2) an increased number of vessels and the presence in syncytiotrophoblast of a wide space near the basal membrane preventing the exchange between the maternal blood and the fetal blood. PMID- 12898872 TI - Collagenoses and cutaneous manifestations of hepar diseases. AB - The authors discuss cutaneous manifestations of some systemic diseases (collagenoses) and their connection with diseases of liver cells. PMID- 12898873 TI - The liver and its ultrasonographic picture in children suffering from vitiligo. AB - The size and the structure of the liver in 38 children with vitiligo and 43 healthy children suffering from a mild form of pityriasis capitis were measured in an ultrasonographic examination and then compared. In the examined group there were found single cases beyond the norm. No changes in the structure of the liver parenchyma were stated in any of the vitiligo cases. For the liver measured in L1 there were stated 94.7% cases of vitiligo within the range from 2.5 to 97.5 percentile values in the control group. For the liver measured in L2--92.5% cases from 2.5 to 97.5 percentile values in the control group and for the liver measured in L3--92.1% cases of vitiligo were within the range 2.5 do 97.5 percentile values in the control group. PMID- 12898874 TI - Changes of resistance parameters of femoral bone in adult female rats after application of zoladex and vincristin. AB - The study consisted in testing the influence of vincristin and zoladex on mechanical resistance of the femur bone in adult female rats. Vincristin was administered intraperitoneally and zoladex--subcutaneously. The experiments imply that in the group of animals receiving vincristin the bone resistance decreased by 6.4% as compared to the control group, and in the group of animals receiving zoladex--by 14.3%. PMID- 12898875 TI - The combined effect of systemic hypertension and obesity on biventricular diastolic function. AB - Left ventricular (LV) cardiomyopathy in hypertensive obese patients is a consequence of increased LV afterload due to systemic hypertension and increased LV pre-load subsequent to the augmentation of total blood volume in obese. However, little is known about right ventricular (RV) functional status and biventricular interaction in patients with systemic hypertension and concurrent obesity. The aim of our study was the echocardiographic assessment of left and right ventricular function and diastolic performance in hypertensive obese patients. The study group consisted of 31 subjects with BMI > 26 kg/m2 and mild to moderate systemic hypertension, patients with systemic hypertension and normal BMI served as a control group. M-mode echocardiographic measurements of left and right ventricles were performed. By pulsed Doppler echocardiography LV and RV recordings of diastolic filling and systolic flows in pulmonary artery were obtained. Hypertensive obese patients demonstrated significantly thicker posterior wall and enlarged left atrium, while no differences in RV structure were found between both groups. In obese subjects we showed significantly lower LV early to late filling wave velocity ratio and LV early to late filling wave velocity-time-integral ratio (p < 0.01). Only RV early to late filling wave velocity-time-integral ratio differentiated obese and lean subjects (p < 0.05). In obese subjects we demonstrated significantly increased mean pulmonary artery pressure. The results of our study indicate that RV is protected from the direct influence of hemodynamic changes observed in overweight/obesity better than the left ventricle and RV abnormalities in patients with systemic hypertension and obesity depend mainly on increased RV afterload, secondary to LV dysfunction. PMID- 12898876 TI - Arthrogryposis multiplex congenita (AMC)--case report. AB - AMC (which can be also described as fetal muscle cicatrisation) is a term describing the presence of various arthrogenic contractures at the moment of birth, limiting, to a certain degree, the range of joints mobility. The incidence of the disease is low--1 per 3,000 births and it is usually not determined genetically. In a 12-year-old patient treated in Paedodontic Dentistry Department of the Medical University of Lublin the contractions were observed in elbow joints, wrist joints and ankle joints. Dental examination revealed microgenia, limited mobility in temporo-mandibular joints and delayed teething. DMF mean value of permanent teeth was 4. Earlier the patient has not undergone any dental treatment. The conservative and orthodontic treatment applied was difficult due to limited mandible abduction (2-3 cm) and limited lateral movements. Children and adolescents with such diseases should undergo intensive prophylactic procedures because their dental treatment is very complicated. PMID- 12898877 TI - Oral hygiene in children with Down's syndrome. AB - The clinical examination comprised 62 children with Down's syndrome: 12 children had low grade mental retardation, 36 children--moderate grade mental retardation and 14 children--high grade mental retardation. The study showed that oral hygiene was unsatisfactory and was found to be dependent upon the grade of mental retardation. Most children had birth weight over 2,500 g. Besides, also birth length was found to be dependent upon the grade of mental retardation. In most families the child with Down's syndrome was the last baby born among other healthy siblings. Educational career in children with Down's syndrome was found to be dependent upon the grade of mental retardation. PMID- 12898878 TI - Regaining functional independence by neurological patients. AB - The aim of the study was to evaluate the effect of nursing care on the process of regaining functional independence by neurological patients. The study covered 80 patients--28 males and 52 females, aged 39-67. The data concerning patients were collected by means of a questionnaire and observation. The results obtained were expressed as a definite number of scores on the Neurological Patient's Functional Independence Scale developed by the author. The study was conducted twice among the same patients. PMID- 12898879 TI - The study of the level of self-care among neurological patients. AB - The assumption of the study was to show the differences in shaping self-care by patients in two different models of nursing: traditional and modern (nursing process). In Group E self-care efficiency increased from the initial value of 10.02 to 31.13 in the third measurement, i.e. over three times, whereas in Group C--from 9.93 to 19.15 i.e., it was nearly twice as high. Only positive effects of changes were observed in both groups which consisted of patients attaining higher categories of self-care. PMID- 12898880 TI - The influence of nonsteroidal anti-inflammatory drugs abuse and chronic stress on the development of necrotizing fasciitis--case report. AB - In this report we present the case of a 19-year-old woman who was admitted to hospital due to an intense pain of the left gluteofemoral region and the symptoms of toxic shock. For 5 days before admission to hospital the patient had a strong pain and fever, up to 40 degrees C, treated with high doses of nonsteroidal anti inflammatory drugs administered by oral and intramuscular routes. During the next 20 hours a large oedema of the left femoral region developed, the skin became purplish, and bullae filled with the hemorrhagic fluid appeared. On the group of typical skin changes, results of computed tomography and findings during surgical treatment necrotizing fasciitis was recognized. The patient was transferred by plane to the National Center of Hyperbaric Medicine Institute of Sea and Tropical Medicine where she was successfully treated with hyperbaric oxygen therapy. In this paper we discuss the problem of the association between the use of nonsteroidal anti-inflammatory drugs, chronic stress and the development of necrotizing fasciitis. PMID- 12898881 TI - The value of spatial reconstructions with computed tomography (3D CT) in recognising the fissure of vertebral arch. AB - The aim of the study is to assess the value of spatial reconstructions with computed tomography (3D CT) in recognising vertebral arch fissure. The material comprises 20 patients in whom 3D CT revealed vertebral arch fissure. Spatial reconstructions at bone threshold were done from computer data of axial sections. 3D CT was assessed in real time in optional projections and lightened at a selected angle. It was found that the most favourable projections for the assessment of vertebral arch fissure are lateral and antero-oblique ones performed from the inside of the vertebral canal after cutting off part of the reconstructed picture along spinous processes. In patients with degree I slip 3D CT in recognising arch fissure is a technique of choice. PMID- 12898882 TI - The value of radiological examination in postoperative diagnostics of cardiospasm. AB - The aim of the study is to assess the value of radiological examination in determining the efficacy of combined operative treatment of cardiospasm. The study group comprised 38 patients, in whom control radiological examinations were performed after surgical treatment of achalasia of oesophagus. The assessment of motor function and efficiency of the cardia was done with physical, radiological examination, oesophagoscopy including taking specimens for histopathologic examination, 24-hour pH-metry and manometry. Different radiological pictures of the gullet and cardia after cardiotomy surgery with subsequent antireflux procedure were presented. It was found that radiological examination in patients surgically treated for cardiospasm enabled the assessment of the efficacy of gullet emptying, width of fundoplication cuff and efficacy of antireflux procedure and that in the assessment of motor function it shows a limited value in correlation with manometric data. PMID- 12898883 TI - Lipid and hormone profile in psoriatic females. AB - The serum concentrations of sex hormones in appropriate days of the menstrual cycle, and of lipids in 15 ovulating women, were measured. The psoriatic patients were compared with a control group of healthy subjects matched for age, sex and body mass index (BMI). In psoriatic patients, the levels of measured hormones were lower than in the controls, with the exception of LH concentrations, which were increased but in a statistically insignificant way. The testosterone levels and LH/FSH ratio were significantly lower in the psoriatic group. The triglyceride concentrations were significantly higher in the patients, and this phenomenon was not related to their body mass. PMID- 12898884 TI - Interleukin-18 serum concentration in patients with psoriasis triggered by infection. AB - The blood serum concentrations of IL-18 and the subpopulation of blood lymphocytes in peripheral blood were investigated in 14 patients with psoriasis preceded by a bacterial infection. The IL-18 concentrations were measured with the ELISA method, and the lymphocyte subpopulation--by flow cytometry. The elevated IL-18 concentrations in blood serum were observed; however, the increase was statistically insignificant in the group of psoriatic patients in comparison with the control (p > 0.4). The result of flow cytometry showed that a statistically significant increase of the percentage of CD45RO4 and CD45RO cells, and a statistically significant decrease of MFI expression of CD19+ and CD3+ cells (p < 0.004; p < 0.02; p < 0.05; p < 0.05) occurred in the patients. PMID- 12898885 TI - Burdensome factors occurring at school according to students. AB - The aim of the work was to find out about school environment and burdensome factors that occur at school according to students. The research was conducted at Primary School no. 23 in Tarnow. 244 students were examined, including 135 students of 8th form and 109 students of 4th form. Troublesome factors, which students are exposed to, may be divided into two groups. The first group is related to physiochemical environment, the second group are the factors which result from a child's way of living and working in the school environment. On the basis of the analysis of the results, the necessity of restriction of both physical and psychosocial burden of students is emphasised. What seems to be especially crucial is ergonomic school furniture, diminishing of noise, improvement of hygienic condition of toilets, rational school timetable, and elimination of dangers resulting from other students' behaviour. PMID- 12898886 TI - Changes in serum concentrations and in cytokine production by blood cell cultures of a patient with major depression--case report. AB - The aim of the paper was to compare the periodical changes in serum cytokine levels and in cytokine production in short-term blood lymphocyte cultures of two persons: the patient with major depression and healthy control. In sera of both persons such cytokines as IL-1 beta, IFN-gamma and IL-6 were detected, but IL-6 level in serum of depressed patient was higher than that observed in healthy control. In both persons examined serum cytokine level changed periodically during 9 days of observation showing rhythmic waves. When cytokines were induced in lymphocyte cultures periodical changes in their production were also observed, but IL-1 alpha, IL-1 beta and IL-12 production was significantly lower in comparison to lymphocytes of healthy control. After 36 days of medical treatment cytokine levels in serum of patient normalized, except for IL-12. Our results suggest that inconsistence in the data from papers of different authors concerning cytokine production in major depression could result from periodical changes in their level and from the fact that patients were examined at the time when certain cytokine was near its peak or declined significantly. The rhythmic changes in cytokine level should be taken into consideration when the role of cytokines in major depression is examined. PMID- 12898887 TI - Anxiety and fear level in patients after myocardial infarction over five years of rehabilitation. AB - The aim of the study was to characterise the structure of anxiety and fear in patients after myocardial infarction observed during their five-year rehabilitation. The study group comprised 114 men aged 24-63 years treated at the Chair and Department of Cardiology, Medical University of Lublin with recent myocardial infarction diagnosed according to generally accepted clinical criteria. Besides basic clinical examinations the patients were performed echocardiography and psychological examination. After completion of the first clinical therapy a five-year post-infarct outpatient rehabilitation was organised for the patients. To assess the level of anxiety and fear Cattell's IPAT Self Analysis was used. The method lets find out that: 1. In every study period the level of general anxiety was close to the neurotic level. 2. During the whole period of five-year rehabilitation in about 60% of patients the normal level of internal integration was observed. 3. In all study periods about 30% of patients showed emotional instability. 4. In the structure of anxiety and fear about 60% of patients always showed high level of inner tension associated with a conflict between superego demands and impulsive sphere. 5. Observation of patients after myocardial infarction lasting many years has shown that the study group is not homogenous as for personality traits. The use of agglomerate analysis allowed distinguishing 7 groups of patients with the most similar personality characteristics. Individual groups of patients significantly differed from one another with regard to the structure of anxiety and fear. PMID- 12898888 TI - The assessment of the 12-year-olds' dental care in rural areas. AB - The following paper presents an analysis of the state of health of the oral cavity in 12-year-olds from the Lublin province. The frequency of dental caries in the examined population as well as its intensity was measured with the mean DMF number, including its particular components. The results of the study revealed a high frequency of dental caries and mean DMF as well as a low treatment index. The results emphasize the need for some intensification of basic dental care among children from rural areas which will concentrate on prophylactics, education and health as well as treatment. PMID- 12898889 TI - Retrospective analysis of mortality rates among the patients with cerebral stroke hospitalized at the Department of Neurology of the Medical University of Lublin in 1990 and 2000. AB - The aim of the study was to assess the mortality rate among the patients with cerebral stroke treated at the Department of Neurology of the Medical University of Lublin. The retrospective analysis included 413 patients hospitalized in 1990 and 2000. The following parameters were evaluated: the type of stroke, age, sex, the length of hospitalization, kinds of complications. The patients' age ranged from 17 to 89 years. The examinations were performed only in the patients with the determined diagnosis of stroke (confirmed clinically, by CT, MRI and anatomicopathologically). These examinations were also used to determine the type of stroke (haemorrhagic, ischaemic). The statistical analysis was carried out using the chi 2 Pearson's test; p < 0.005 was assumed to be statistically significant. The findings reveal a statistically significant decrease in the mortality rate among the patients with haemorrhagic cerebral stroke in 2000 compared to 1990. The mortality rate in the ischaemic stroke patients was also reduced, however the result was not statistically significant. PMID- 12898890 TI - Histological examination of the lacrimal gland after experimental administration of Cladribine. AB - The experiment was carried out on female rabbits of New Zealand breed weighing about 3 kg. The rabbits from experimental group I received Cladribine in the dose corresponding to the schema of treatment in the hairy cell leukemia and the animals from the experimental group II the dose corresponding to the experimental treatment in multiple sclerosis. The lacrimal glands were collected for histological examinations in the light microscope. It was observed that administration of Cladribine in the dose corresponding to the therapeutic dose used in the therapy of hairy cell leukemia could have caused morphological changes in the lacrimal gland, but the medicine administered in therapeutic doses in experimental therapy of multiple sclerosis does not cause morphological changes in the structure of the lacrimal gland on the level of the light microscope. PMID- 12898891 TI - The occurrence of primary open angle glaucoma in a family. AB - The occurrence of primary open glaucoma (POAG) in families proves the hypothesis that POAG is a disease with underlying genetic factors. This work describes a family of four generations, in which eight members suffered from POAG. In the 3rd and 4th generation members, the disease occurred at an early age (between the age of 20-40 years), i.e. at a much earlier age than it could be expected. Therefore, a need arises to keenly observe families in which POAG is present. PMID- 12898892 TI - Hospital urinary tract infections in surgical patients. AB - The subject of the present study was evaluation of the prevalence of the urinary tract infections (UTI) in surgical patients, as well as determination of the type and susceptibility of bacterial flora responsible for the UTI. The UTI constituted 21% of all hospital infections in the surgical ward and involved 2.3% of all operated patients. The main etiological factor was Gram-negative bacteria, which were responsible for 82.3% of infections. These infections were characterised by high susceptibility to carbapenems, the 3rd generation of cephalosporins, and fluorochinolons, as well as marked resistance to nitrofurantion derivatives, aminoglycosides, second generation of cephalosporins, and amoxicillin with clavulanic acid. Over 21% of Gram-negative bacteria produced beta-lactamase of extended spectrum (ES beta L). Moreover, 94% of recognised hospital UTI were associated with previous catheterisation of the urinary bladder. PMID- 12898893 TI - Risk factors of infection in surgical wounds. AB - The article deals with the problem of surgical wound infections in patients of surgical wards. Modern definition and etiology of surgical wound infections in Poland and elsewhere have been presented here. At the same time present opinions referring to the problem of surgical wound infections have been discussed. PMID- 12898894 TI - The influence of application of 3D CT reconstructions on classification of maxillofacial fractures. AB - The aim of the paper was to study of the influence of application of three dimensional CT reconstructions on diagnostics and classification of maxillofacial fractures. The material comprised 97 patients presenting with maxillofacial trauma who underwent CT examination in the 2nd Department of Medical Radiology of the Medical University of Lublin in the years 1995-2001. The CT examinations were performed using the Somatom AR-T machine in coronal and axial slices, 2 and 3 mm thick. Then the 3D CT reconstructions were obtained. All cases of trauma were classified according to Wanyura first on the basis of radiograms and CT examination, next on the basis of 3D CT reconstructions. The results of the two analyses differed in cases of properly diagnosed zygomatico-orbital fractures and orbitonasal dislocations, which were less numerous during the first reviewing. It was found that 3D CT reconstructions were generally useful in classifying the maxillofacial fractures with the exception of isolated orbital floor fractures. PMID- 12898895 TI - Tumour's angiogenesis--the function of VEGF and bFGF in colorectal cancer. AB - The main objective of this paper was to present the view on the vascular endothelial growth factor (VEGF) and basis fibroblast growth factor (bFGF) in the angiogenesis in the colorectal career. The analysis includes the works of these scientists, who proved the relationship between VEGF and bFGF expression and the advancement level of colorectal cancer and the survival of the patients ill with this disease. It was stated that the highest levels of these factors were connected with the advancement of neoplastic process, especially when metastases coexisted at the same time. It was also proved that the higher levels of VEGF and bFGF gave worse prognosis as far as survival was concerned. Apart from this, other factors effecting angiogenesis and inhibiting factors were also presented. PMID- 12898896 TI - The correlation of US and CT values in the diagnostics of giant pseudo-cysts of the liver. AB - The aim of the study was a comparative assessment of the value of US and CT examinations in the diagnostics of giant hepatic pseudo-cysts. The material comprises 15 cases of complex hepatic pseudo-cysts with diameter exceeding 5 cm, which in 6 cases had traumatic etiology, in 4 neoplastic (cystoadenomas and cystoid adenocarcinomas) and in 5 cases infectious (postinflammatory and parasitic). Considerable disproportions in US and CT pictures of liver cysts were revealed. Posttraumatic cysts showed smooth inner contours in CT while in US irregular contours with thin septa intussuscepting to the inside were found. In 2 cases of neoplastic cysts thin inner septa were only shown with US examination. In parasitic cysts CT visualized only a part of septa visible in US. It was found out that considerable disproportions of US and CT pictures of complex hepatic cysts require combination of both imaging methods in their diagnostics. US shows high sensitivity in revealing septa, determining internal contours and wall thickness. CT reveals calcifications and after contrast enhancement vascularised walls, septa and tissue component. CT is useful in determining topographic relations of big hepatic cysts. PMID- 12898897 TI - The effect of chronic alcohol abuse on gastric and duodenal mucosa. AB - Alcohol consumed in small quantities is not dangerous for health but if it is drunk in big amounts it has a negative effect on somatic and psychical health. A number of studies have been published on the harmful effect of ethanol on the nervous system, circulation, endocrine and immune systems. Ethyl alcohol can directly damage the mucosa of the alimentary tract. The studies conducted and published so far have estimated acute ethanol damage of gastric mucosa in experimental animals. The views found in literature on the effect of continuous consumption of ethanol on the stomach are differentiated. The purpose of the paper was to provide a morphological and functional analysis of stomach and duodendum in patients who continuously abuse alcohol. Besides, at attempt was made to find the answer to the question whether the changes observed in gastric mucosa point to any connection with the period of alcohol abuse. The study referred to 79 persons: 61 male aged 17-65 who had been chronically drinking alcohol for the period of 5-37 years and 18 persons aged 17-59 (9 male and 9 female) who had never drunk alcohol or who had drunk it only rarely and in small were divided into three groups differing with the period of addiction (I--5-10 years; II--10-20 years; III--over 20 years). Hydrochloric acid secretion in gastric juice was marked and gastroscopy was performed in all the examined patients. Gastroscopy estimated the appearance of the gastric and duodenal mucosa, while biopsy specimens were taken for histological examination from the pyloric area, the body of the stomach and the duodendal bulb. Hydrochloric acid secretion was determined using the Kay's aspiration method. Pentagastrin was used to stimulate gastric secretion. Within the group of 79 patients, gastric mucosa inflammation was observed in all patients chronically drinking alcohol and in 72% patients who were the control. Both groups differed considerably with the degree of progression of the inflammatory process, which was estimated with histological examination. Atrophic gastritis was observed only in patients addicted to alcohol. The appearance of atrophic changes pointed to a close relation with the period of addiction. 13 out of 14 people with this type of inflammation had been drinking alcohol for at least 10 years. The studies did not show any relation existing between the percentage of cases with atrophic inflammation and the kind of drinks or the content of ethanol in them. Examinations concerning the secretory function of the stomach showed lower values of hydrochloric acid secretion, both in basic conditions and after pentagastrin stimulation, in patients addicted to alcohol as compared to the control. The results prove that continuous abuse of alcohol predisposes to atrophic inflammation of the gastric mucosa, and the appearance of this type of inflammatory changes is related to the duration of addiction. The longer the addiction, the lower the secretion of hydrochloric acid is. PMID- 12898898 TI - The promotion of happiness. AB - The human mind is capable of creating an internal world--the psychic sphere- including the phenomena characteristic of human nature, such as selfconsciousness, conscious experiences, conceptual thinking, symbolic language, dreams, art, creation of culture, sense of values, interest in the distant past and care about the distant future. According to the exceptionally concordant opinions of the sages and scholars of the East and the West it is just within this internal world that human happiness dwells. Happiness is a state of the spirit which consists in: internal peace, satisfaction with one's life, the joy of life, benevolence and cordiality towards oneself and towards others; sensitivity to the beauty of nature, culture and art; harmonious co-existence with the surroundings. The achievement and experiencing of the states of thus understood happiness depend mainly on ourselves and, similarly to the project of health promotion, require knowledge, willingness and possibilities. Happiness Promotion denotes the commendation and popularizing of a certain definite way of thinking and acting--showing the road which leads to the frequent experiencing of happy moments. PMID- 12898899 TI - Does ACE inhibitor modify the morphofunctional state of the nephron? AB - Captopril, the inhibitor of the angiotensin convertase (ACE) was administered to white rats in two doses: 0.23 mg/day/rat and 0.71 mg/day/rat. The preparation was being applied for the period of 3 weeks. Observations were carried out in a transmission electron microscope. In the kidneys of the animals subjected to the experiment, in the cells of the proximal canaliculi dilution of the cytoplasm, numerous vacuoles in the apical and basal part of the cell, dilation of the intercellular space as well as more numerous lysosomes were observed. The above described changes were more distinctly marked in animals which were given a higher dose of the drug. The studies in the electron microscope revealed morphofunctional changes of the nephron related to the application of the Captopril. Application of the Captopril may temporarily modulate the morphofunctional status of certain structures of the nephron, which was demonstrated in the foregoing experiment. PMID- 12898900 TI - Comparison of selenium distribution in mice organs after the supplementation with inorganic and organic selenium compound selenosemicarbazide. AB - Studies on selenium organ content and its function in living organisms just like studies on other elements provide interesting results although their interpretation is not always clear. The aim of our study was to determine the concentration and distribution of selenium in several organs and tissues in mice after supplementation with our newly synthesized organic compound of selenium selenosemicarbazide (4-o-tolyl-selenosemicarbazide of o-chlorobenzoic acid) as compared to the effects of the supplementation with inorganic compounds. SWISS mice were fed with both types of compounds at the dose of 10(-3) g Se per kg for the period of 10 days. The concentrations of selenium in brains of mice treated with selenocarbazide and sodium selenite were higher than in controls (38.04 micrograms g-1 and 32.00 micrograms g-1 vs. 26.18 micrograms g-1). There was a statistically significant increase in the selenium contents in lungs after supplementation with selenosemicarbazide and sodium selenite (11.81 micrograms g 1 and 6.79 micrograms g-1 vs. 1.75 micrograms g-1 in controls). We found a statistically insignificant increase in selenium contents in intercostal muscles after supplementation with inorganic selenium compounds and a statistically significant increase after the supplementation with selenosemicarbazide (10.13 micrograms g-1; 14.21 micrograms g-1 and 28.84 micrograms g-1, respectively). Our investigations lead to a conclusion that 4-o-tolyl-seleno-semicarbazide of o chlorobenzoic acid, an organic selenium compound may be more easily absorbed than inorganic sodium IV selenite. PMID- 12898901 TI - Prognostic value of the atrial pulsed Doppler recordings of transmitral flow in the assessment of left ventricular diastolic dysfunction. AB - Volume sample location is the most important factor influencing pulsed Doppler recordings of left ventricular (LV) diastolic filling. Transmitral diastolic flow velocities measured at the tips of mitral valve leaflets are usually higher than those obtained from atrial location. Nevertheless, independently of volume sample location, differences between early to late velocities ratio (E/A) are nonsignificant, so E/A became the most useful index of ventricular diastolic performance. In some patients, however, with E/A > 1 measured between the tips of mitral leaflets, the pattern of delayed LV relaxation, with diminished E and predominant A wave (E/A < 1), is often obtained from left atrial volume sample location. The aim of our study was to compare LV dimensions and wall thickness between patients with E/A > 1 recorded both at the tips of mitral valve leaflets and in the left atrium and patients with E/A > 1 at the tips of mitral valve leaflets and E/A < 1 in the left atrium. The study population consisted of 51 patients with E/A > 1 recorded between the tips of mitral valve (E/Avent) and excluded pseudonormalized filling pattern. The study population was divided into two groups based on Doppler profile recorded in the left atrium: group A--26 patients with E/A < 1 (E/Aatr) and group B--25 patients with E/Aatr > 1. Patients from group A were significantly older (55.6 +/- 12.8 vs 43.5 +/- 10.5 yrs, p < 0.001) and had increased body mass index (28.8 +/- 4.4 vs 25 +/- 4.1 kg/m2, p < 0.01) compared to subjects from group B. The average thickness of intraventricular septum was 13.5 +/- 2.8 mm in group A and 9.7 +/- 1.6 mm in group B, the average thickness of LV posterior wall was 11.6 +/- 1.7 and 9.2 +/- 1.8 mm, respectively. The results of this study suggest that abnormal atrial recordings of diastolic flows may reflect the earliest stage of left ventricular diastolic dysfunction in asymptomatic patients with echocardiographic signs of mild cardiac damage. PMID- 12898902 TI - Comparative analysis of the spirographic and hemodynamic parameters in preoperative evaluation of patients with lung cancer. AB - Surgery is the recommended treatment for nonmicrocellular lung cancer. The spirometric tests completed with gasometric systemic blood analysis and electrocardiographic investigation are the standard in the evaluation for tolerance of lung tissue resection. The catheterization of the right heart is used to assess circulatory systems efficiency in patients qualified for pulmonary tissue resection. The purpose of the study was to evaluate the correlations between basic spirographic and hemodynamic parameters measured at rest and after exercise. The studied population consisted of 50 consecutive male patients 21 to 72 years old with bronchial carcinoma, considered to be candidates for lung tissue resection. The vital capacity, and one-second forced expiratory volume were obtained. The hemodynamic examination was performed using Swan-Ganz thermodilution catheter. The systolic, diastolic and medium central venous pressure, pulmonary artery pressure and pulmonary wedge pressure were measured. Gasometric examination for peripheral arterial and mixed venous blood was performed--pH, PO2, PaCO2, SaO2. The results were subject to statistical analysis. The correlation between spirographic indicators and hemodynamic parameters were studied. Statistical analysis revealed high level of correlation between the average blood pressure in pulmonary artery, both at rest and after exercise, and ratio of one-second forced expiratory volume related to current vital capacity. The revealed correlations allow to solve regression equations making in possible to calculate likely values of the average blood pressure in pulmonary artery on the basis of spirographic values. PMID- 12898903 TI - Comparative analysis of the gasometric and hemodynamic parameters in preoperative evaluation of patients with lung cancer. AB - Measurements including spirographic examination, arterial blood gas tensions, standard electrocardiogram are routinely used to define risk factors for patients undergoing lung tissue resection. In some instances routine functional check-ups should be accompanied by the assessment of the hemodynamics of pulmonary circulation. The purpose of the study was to evaluate the correlations between gasometric and hemodynamic parameters measured at rest and after exercise. In case of the presence of such correlations I wanted to find the principles to provide pulmonary artery hypertension in order to avoid right heart catheterization. The gasometric parameters in systemic and in mixed venous blood (pH, PaO2, PaCO2, SaO2) as well as hemodynamic parameters of pulmonary circulation (PAP, PCWP, CVP) were measured in 50 male patients with lung carcinoma. All measurements were taken at rest and after an exercise test--5 minutes, 50 W workload on cycle ergometer in supine position. CI, PVR and SVR were calculated. The study proved statistically significant correlations between gasometric and hemodynamic parameters and made possible to calculate the regression lines equations, which help to predict pulmonary artery pressure before tissue resection. PMID- 12898904 TI - Diagnostic value of electrocardiographic investigations in preoperative evaluation of patients with lung cancer. AB - The pulmonary hypertension and cor pulmonale in patients before lung tissue resection must be predicted even if invasive studies are required. The purpose of the study was to evaluate the diagnostic value of electrocardiographic investigation for patients with lung cancer, considered to be candidates for lung tissue resection. The study comprised 50 male patients with bronchial carcinoma before lung tissue resection. All of them had gasometric, and hemodynamic parameters of pulmonary circulation assessed at rest and after exercise. The ECG and spirographic investigations were done in all patients. It was found that 23 of 50 patients had electrocardiographic traits of right ventricle hypertrophy and only one of them had normal pulmonary circulation parameters. There were also found correlations between ECG curve and FEV1%, most of gasometric and some of pulmonary hemodynamic parameters. PMID- 12898905 TI - The influence of interferon alpha on the rat liver injured by chronic administration of carbon tetrachloride. AB - Due to their complex and not fully known etiopathogenesis as well as difficulties in treatment, chronic hepatitis and cirrhosis still remain one of the main problems of hepatologists. Nowadays, the use of IFN alpha is considered the most effective method of treatment in chronic hepatitis. Recently, a new property of IFN, i.e. its effects on the reduction of fibrosis, has been discovered. The aim of the paper was to examine the effects of IFN alpha on biochemical parameters (AlAt and AspAt activities), on the metabolic function of the liver and its morphologic picture observed under the light and electron microscope after the 3- and 6-week CCl4-induced damage. The experiments were carried out in Wistar male rats. To evaluate the liver function, the test of aminophenazone elimination in the isolated perfused rat livers was used according to Miller modified by Hafte. Additionally, AspAt and AlAt activities were determined. The liver specimens were analysed under the light and electron microscope and using immunohistochemical methods. The findings show that after the 3-week CCl4-induced liver damage, IFN alpha does not significantly affect AlAt and AspAt activities, irrespective of the dose used. IFN alpha administered after the 6-week damage significantly changes those activities when the doses used are high. It was found that carbon tetrachloride does not result in evident cirrhotic changes, however it activates Ito cells, causes focal retraction of the stroma and fibrosis. The increased number of Ito cells in Disse's space observed in immunohistochemical and ultrastructural examinations is indicative of the activation of liver fibrotic processes following CCl4 administration in both variants used. IFN alpha substantially weakens fibrogenesis of the CCl4-damaged liver which is visible in the decreased number of Ito cells and weaker expression of the stroma retraction. Moreover, IFN alpha administered to the experimental animals after the CCl4 induced injury of the liver increases aminophenazone clearance, especially when used in higher doses. Positive effects of IFN confirmed in the studies suggest that the drug may be used in patients with chronic hepatitis and early cirrhosis since it is likely not only to eliminate the virus but also to improve the liver function and reduce fibrosis. PMID- 12898906 TI - Ultrastructural changes of ciliary epithelial nucleus in experimental diabetes- an animal model. AB - The aim of this study was to observe in electron microscope the New Zealand rabbits ciliary epithelial nucleus structure changes, depending on the duration of experimental diabetes mellitus. The animals were administered alloxan in the form of 10% solution in 0.9% NaCl, at the dose of 100 mg/kg b.m. If the glycemia level was 11 mmol/l or higher the animal was included in the experimental group. 15 eyes of 15 adult New Zealand rabbits were studied. Conventional electron microscopy was used to show ultrastructural changes of ciliary epithelium nucleus. During the study we noticed severe changes in the structure of ciliary epithelial nucleus depending on diabetes mellitus duration. We found: deep indentations of nuclear membrane, open nuclear pores, irregularity in heterochromatin location and small sizes of the epithelial nucleus. PMID- 12898907 TI - The influence of baclofen on reflex circulatory reactions evoked by stimulation of the vagus nerve in the rabbit. AB - The purpose of our investigations was to investigate the effect of baclofen upon the reflex fall of arterial blood pressure and heart rate evoked by stimulation of bilateral vagus nerves. The experiments were carried out on 30 rabbits of both sexes and mixed breed weighing 3000-4500 g under urethane anaesthesia (1.5 g/kg). Mean arterial pressure and heart rate were measured in the ear middle artery by the electromanometer and registered by PC computer. Parameters of stimulation were: frequency 5c/s., duration-20 s., width of single rectangular pulse-1 msec., intensity of stimulation--multiplicity of threshold excitation (T). The intervals between stimulations were not shorter than 5 min. Baclofen was administrated intravenously (1.0 mg/kg). In the control group the facilitation of circulatory reaction was observed only at the smallest intensity of stimulation (1.0T-1.5T). When greater intensity of stimulations was applied the sum of unilateral effects was greater than the effect of simultaneous bilateral vagus nerve stimulation. This occlusion effect increased up to the stimulus value of 4-5T. After baclofen administration the facilitation of circulatory responses disappeared and only occlusion was observed. Baclofen increased the size of the area of active overlapping of the afferent innervation of the vagus nerve. Administration of baclofen produced an increase in mean arterial blood pressure (15 +/- 6 mm Hg) and heart rate (16 +/- 5 beats per minute) and decrease in the vagal reflex fall of mean arterial pressure and heart rate. Stimulating of the GABAB receptors markedly attenuates the baroreceptor reflex resulting in increase in mean arterial blood pressure and heart rate. These results support the hypothesis that GABA acts tonically on GABAB receptors to attenuate the baroreceptor reflex, thereby contributing to the regulation of circulatory parameters. PMID- 12898908 TI - Emotional responses of patients with schizophrenia to their illness. Attitudes of social environment towards schizophrenes. AB - The aim of this paper was to define emotional responses of the patients with diagnosed schizophrenic psychosis to their illness. It was also intended to recognize the attitudes of patients' families and more distant social environment towards them. The study was conducted on 84 patients of the Neuropsychiatric Hospital, both hospitalized and ambulant, treated in Mental Outpatient Clinic and, simultaneously, taken care of by Social Self-Help Home "Misericordia". All the examined patients met the diagnostic criteria in accordance with ICD-10 for schizophrenic psychosis or schizophrenic disorder; all were in the period of symptomatic remission. The study was carried out in the years 2000-2001, using the distributed inquiry questionnaire technique. The supplementary source of information was case records of the examined patients. The predominant feelings of schizophrenics at the moment of becoming aware of having fallen ill with mental disease were fear (anxiety) and sorrow. The passage of time caused changes in emotional responses to mental illness. At the moment of the study the predominant feelings were the acceptance of illness and the sense of inferiority because of it. The sufferers of schizophrenia experienced mainly sympathy, acceptance of the illness and indifference in the environment outside the family circle. PMID- 12898909 TI - Changes in epidemiology of viral hepatitis in Poland at the end of the 20th century. AB - Viral hepatitis is, apart from influenza, the most often registered infectious disease in the world. The material comprised epidemiological data concerning the number of cases of viral hepatitis registered in Poland in the years 1990-2000. There were analysed the number of cases, incidence, number of deaths and mortality from viral hepatitis of different types in Poland in the years 1990 2000. The incidence rates from all types of viral hepatitis have been increasing in Poland until 1993 and decreased abruptly after 1993, mainly due to the diminution of number of cases of viral hepatitis type A. There was registered a decreasing trend in incidence rates of viral hepatitis type B that was significantly intensified after 1993, i.e. since the year when mass prophylaxis vaccinations have been initiated. Since registration of cases of viral hepatitis type C was started in 1997, the number of analytically confirmed cases of this disease has doubled. Mortality from all types of viral hepatitis has decreased twofold in Poland in the years 1990-2000. PMID- 12898910 TI - Diagnostic imaging of ameloblastoma. AB - The value of various radiological techniques in cases of ameloblastoma was discussed on the basis of analysis of radiograms and the results of diagnostic imaging methods of patients examined in the Medical University of Lublin in the years 1995-2000. It was concluded that diagnostic imaging of ameloblastoma requires both conventional radiograms and modern imaging techniques such as computed tomography and magnetic resonance imaging. It is imperative to correlate the results of computed tomography and magnetic resonance imaging in cases of recurrence. PMID- 12898911 TI - Radiodensitometric measurements in cases of chronic periapical changes of endodontically treated teeth. AB - The purpose of the paper was radiodensitometric evaluation of hard tissues of roots and alveolar bone of endodontically treated teeth with clinically detected chronic periapical inflammatory changes. The material consisted of 139 digital intraoral radiograms of endodontically treated teeth with chronic periapical changes: 54 teeth with granulomatous inflammation, 62 with fibrous changes and 23 teeth with purulent lesions. There was discussed the value of digital radiography and radiodensitometric measurements in cases of periapical inflammations of endodontically treated teeth. It was concluded that radiodensitometric measurements allowed detailed examinations of inflammatory changes at the apices of endodontically treated teeth. Digital radiography is a valuable and objective tool in diagnosis of periapical changes, monitoring of the course of endodontic treatment as well as in follow-up after such treatment. At the same time it allows reduction of ionising radiation dose, thus decreasing diagnostic risks to a patient. PMID- 12898912 TI - Optimisation of conditions of visualisation of dental crowns in dental radiography. AB - Digital X-ray imaging systems are an alternative to conventional intraoral radiograms. In the paper there are discussed the possibilities of application of different digital radiography tools as well as density measurements in optimization of diagnostics of pathological lesions of teeth crowns. There were analyzed 1,491 digital intraoral radiograms obtained in 985 patients in the Digora Soredex digital radiography system. On all taken X-ray pictures there were evaluated the crowns of teeth in cases of primary and secondary caries, dental fillings, internal resorption, traumatic lesions of incisors as well as the relationships between deep carietic lesions and pulp horns as well as root furcation. An attempt was made to assess the possibilities of differentiation of cervical caries and cervical burn-out. In the paper there were presented the optimal options of digital radiography software in diagnostics of the discussed pathological lesions. PMID- 12898913 TI - Studies on angiogenesis in the benign and malignant ovarian neoplasms with the use of color and pulsed Doppler sonography and serum CA-125, CA-19.9, CA-72.4 and vascular endothelial growth factor measurements. AB - The aim of the study was to investigate whether a combination of serum CA-125, CA 72.4; CA-19.9 and VEGF levels along with several currently used gray-scale criteria and pulsed and semiquantitative color Doppler blood flow assessment can be useful in preoperative discrimination of ovarian tumors. Ultrasound examinations were performed between 1994 and 1999 with the use of transvaginal probes 5-6.5-7.0 MHz and transabdominal 3.5 MHz probe (B&K Medical 3535 and 2002 ADI, Denmark). Preoperative serum CA-125 and CA-19.9 (Roche, Switzerland) and CA 72.4 (Centocor, USA) were measured with EIA II immunoassays. VEGF (R&D Systems, USA) serum levels were measured by microplate immunoenzymatic method. Retrospective study included 4,876 women referred for sonography of pelvic structures. For final analysis 451 patients with persistent adnexal tumors were available. In all these women age, menopausal status, tumor size, volume and morphology as well as blood flow indices (PI, RI, PSV) and semiquantitative color assessment were noted. In women who were not operated the cysts were followed for 10-12 weeks until resolution which was confirmed by repeated sonography. Multiple regression analysis was used to determine which of the independent variables had prognostic significance in the constructed predictive model. ROC curves were plotted and areas under ROC for each test were calculated and compared. 286 women were operated and in 118 patients their tumors regressed. The sensitivity for Doppler indices was between 85% (RI), 67% (PSV) and 66% (PI). For the subjective color assessment sensitivity was 87%. 294 women had their CA-125 serum levels available. For CA-19.9, CA-72.4 and VEGF the corresponding numbers were 109, 92 and 116 patients, respectively. Optimal cutoff level for CA-125 was 67.5 U/ml and this produced diagnostic accuracy of 69.4%. For CA-19.9 the best predictive value was 13.4 U/ml (accuracy of 74%) and for the cut-off CA-72.4 level of 4.1 U/ml the accuracy of the test was 77.8%. Multiple regression analysis revealed that only RI measurements and subjective color assessment had significant influence on the constructed predictive model. The best cut-off value of regression index "Z" was obtained following ROC curves construction for sensitivity (true positive rate) and 1-specificity (false positive rate). In all 116 patients who had Doppler indices with both CA-125 and VEGF levels measured the highest accuracy was associated with "Z" = 1.45 in postmenopausal women. The prognostic model proposed in this study can be log transformed and further used in the estimation of the true probability of malignancy of a given mass in the new set of prospectively studied patients with ovarian tumors. Color and pulsed Doppler can improve preoperative diagnosis of adnexal tumors when compared to transvaginal sonography alone or tumor markers assessment. The proposed semiquantitative evaluation of tumor vascularity increases the predictive value in terms of sensitivity and specificity. VEGF serum concentration was not useful in the preoperative discrimination of malignant and benign ovarian tumors. PMID- 12898914 TI - Influence of time period of cadmium intoxication on the concentrations of ascorbic acid selenium in certain tissues and blood serum of rats. AB - Ascorbic acid and selenium are important antioxidants for the reactive forms of oxygen. The purpose of these experiments was to determine the influence of various concentrations of cadmium on ascorbic acid and selenium concentrations in certain tissues of rats. The results showed that cadmium, added to rats' feed, influenced the concentration of ascorbic acid in blood serum and tested rats' tissues. Cadmium influenced the concentration of selenium in certain tissues as well. Furthermore, it has been demonstrated that ascorbic acid and selenium concentrations are dependent on the type of tissue tested as well as on the size of cadmium concentrations and length of the experiments. PMID- 12898915 TI - The influence of zinc and copper supplementation in feed on the concentrations of certain metals in rats' skin. AB - Zinc and copper, administered to the organism in excess can also influence the concentrations of other elements, and have a secondary influence on the processes that these elements regulate. The purpose of this experiment was to determine the influence of zinc and copper dietary supplementation on the concentrations of magnesium, calcium, zinc and copper in the rat's skin. The results showed that zinc supplementation caused a decrease in calcium concentration as well as an increase in zinc concentration in the tested materials. The concentrations of magnesium and copper changed insignificantly. Copper supplementation caused a decrease in the concentration of magnesium, calcium and zinc but caused an increase in copper concentration. Changes in concentrations of the tested metals were dependent upon the concentrations of various metals added to the rat's diets. PMID- 12898916 TI - Blockade of reticular formation activity, due to carisoprodol maternal administration, and its effects on rat skeleton development. AB - The purpose of this experiment was to study the influence of reticular formation blockade, due to carisoprodol maternal administration, on rat skeleton development. The drug was administered three times a day orally by stomach tube at doses: T1--20 mg/kg/24 h, T2--200 mg/kg/24 h, T3--400 mg/kg/24 h. The fetuses obtained on 21st day of gestation were counted and macroscopically examined. Placental and fetal weight, fetal and tail length were checked. After fixation in 95% ethanol the fetuses were stained under single alizarin red S Dawson method and examined under a stereo-dissection microscope. Morphological examination revealed no major malformations. Insignificant number of subcutaneous ecchymose and various skeleton anomalies were observed. The experiment revealed that carisoprodol has no influence on rat skeleton development. PMID- 12898917 TI - Desmopressin--a nontransfusional form of prevention of postextractive bleeding in some patients with von Willebrand's type I disease. Case report. AB - In this article the possibility of using the Minirin (desmopressin) in maxillofacial surgery in person with hemostatic disorders has been described. The our case of patient with von Willebrand's disease type I indicates that this drug is very effective in bleeding after teeth extractions. PMID- 12898918 TI - Pseudoductules in the rat liver in experimental adriamycin-induced nephrotic syndrome. AB - Pseudoductules proliferation in the liver is the condition observed in some liver diseases in human and experimental animals. It is observed in viral and alcoholic hepatitis, in hepatic cirrhosis and toxical liver damage. In that study pseudoductules were observed after i.p. single dose of adriamycin (5 mg/kg of body weight) administered to rats. Adriamycin given in this way is used to induce experimental nephrotic syndrome. PMID- 12898919 TI - Efficacy of the triple therapy: proton pomp inhibitors, amoxicillin and tynidazole in Helicobacter pylori infection treatment. AB - The aim of the paper is to present the triple therapy results combined with the application of proton pump inhibitors with amoxicillin and tynidazole in treatment of the contagion caused by Helicobacter pylori. The therapy was carried out among 108 people after realization of endoscopic biopsy and with quick urease test. In the endoscopic investigation 72 people were found to have duodenum ulceration. Inflammation of mucous duodenum was found in 36 patients. Among 108 people under investigation 51.85% were men and 48.15% women. These people were aged between 15-81 (the average age was 44). After the use of eradication 99 out of 108 patients (91.66%) were cured. The lack of improvement was found in 9 people (8.34%). The age of these people varied from 18 to 75, which gives the average age 54.5 (this age is about 10 years and a half higher in relation to the average age of the examined group of patients). The results prove high efficiency of this schema of treatment in relation to comparatively low costs. They also indicate greater efficiency of the treatment of contagious caused by Helicobacter pylori in younger people. PMID- 12898920 TI - Histological changes in the pancreas of white rats after administering cephalexin and ethanol. AB - The research was conducted on male Wistar rats weighing approximately 200 g. Animals of experimental group I were administered 20% ethyl alcohol for drinking, animals of experimental group II--cephalexin in the dose of 42 mg daily, animals of experimental group III--simultaneously alcohol and cephalexin in the mentioned doses. After 10 days the animals were guillotined and pancreas was taken for research. On paraffin sections 7 mu thick there were carried out H + E stain and PAS reaction aimed at discovering neutral mucopolysaccharides. After administering alcohol there was stated a decrease in the activity of exocrime cells and after administering of cephalexim--an increase in this activity. A simultaneous administration of ethyl alcohol and antibiotic causes trophic changes, which can be noticed as introductory, but at this stage--reversible, degenerative changes. PMID- 12898921 TI - Histological examination of the submandibular gland following experimental administration of Metizol. AB - The submandibular gland of the white Wistar rats was examined. The animals were given Metizol for 21 days and 42 days at the dose of 1 mg/kg b.m./24 h. The submandibular gland samples were taken for histological and histochemical examination. Then they were stained with hematoxylin and eosine as well as by Masson's, PAS's and Feulgen's method. The mean area of section of cell nuclei was measured. The results of examination were counted statistically. The following changes were noticed: After 21 days of administration of Metizol in the submandibular gland the mean area of the tubules was increased. The quantity of tubules increased as well. In the tubules cells some more secretion was noticed. The follicles shrank. After 42 days of administration of Metizol the appearance, number and stainability of the tubules and follicles were similar to control group. PMID- 12898922 TI - Histological examination of the Loeventhal gland after experimental administration of Metizol. AB - The Loeventhal gland of the white Wistar rats was examined. The animals were given Metizol for 21 days and 42 days at the dose of 1 mg/kg b.m./24 h. The Loeventhal gland's samples were taken for histological and histochemical examination. Then they there stained with hematoxylin and eosin, Masson's, PAS's, and Feulgen's method. The mean area of section of cell nuclei was measured. Results of examination were counted statistically. The following changes were noticed: after 21 days of administration of Metizol in the Loeventhal gland the mean area of the section of cells nuclei was decreased; after 42 days of administration of Metizol the mean area of the section of cell nuclei was decreased as well, but to a lesser degree than in group 1. New follicles appeared which can be the expression of cell mitotic activity. PMID- 12898923 TI - The influence of chronic obstructive pulmonary disease on the occurrence rate and intensification of osteoporosis. AB - The aim of this study was the estimation of the relationship between osteopenia and osteoporosis and chronic obstructive pulmonary disease (COPD). The group under investigation consisted of 77 people suffering from chronic obstructive pulmonary disease in different stages of its development and control group. All the patients underwent spirometry carried out by means of abcPneumoRS system and bone densitometry carried out by means of Osteometer A/S using the DEXA method; the results were compared with those of the reference groups. Static and dynamic lung volumns were measured in all patients. By means of densitometry the mass, bone mineral density and T-score were determined in all patients' distal forearm. The research confirmed the relationship between the mass and density of bones and patients' age, sex, weight and body surface. Osteoporosis frequency rate was much higher among patients with chronic obstructive pulmonary disease, particularly women. The following relationship between the degree of disease progression and the mass and mineral density of the bones was observed: the more advanced airflow obstruction was, the lower osseous mass coefficients were. Forced expiratory volume during the first second and airflow obstruction reversibility rate determined by means of bronchodilator response also had a considerable influence on osseous mass coefficients. PMID- 12898924 TI - The value of health in the opinion of patients after myocardial infarction. AB - Health as the counter-value of disease is a vital value. The disease as a stressful situation affects the existing attitudes of man and his relations with the environment. This is as opportunity for changing the hierarchy of values and mobilising own creative potential. The study aimed at determining the attitudes of people after myocardial infarctions towards the value of health. Investigations were performed in 1999 in a group of 250 persons, who had had heart attack in the previous 4 years' period. Study results indicate that health is declared as the value "most important of all most important ones" and the most desirable in the category of "life goals endeavours". A stronger than earlier tendency is also observed in the situation of chronic disease for undertaking wholesome efforts. Generally, however, there is still found too small dedication to create a healthy lifestyle, both on the level of knowledge and skills and motivation necessary for carrying it out. PMID- 12898925 TI - The level of alpha-antitrypsin and ceruloplasmin in the subretinal fluid of patients with rhegmatogenous retinal detachment. AB - The aim of the study was to determine alpha-antitrypsin (AAT) and ceruloplasmin (CER) level in the subretinal fluid of patients with rhegmatogenous retinal detachment (RRD). Investigation was performed in 41 patients (22 women, 19 men) operated on for RRD at the age from 24 to 81. The group of patients was divided with regard to the extent and duration of RRD. In 24 patients detachment involved 1 or 2 quadrants and in 17 patients--3 or 4 quadrants. In 16 subjects RRD lasted shorter than 2 weeks, in 6 patients--about one month and in 19 patients--from 1 to 3 months. The subretinal fluid was collected during operation. The levels of the examined substances were determined with the radial diffusion method on Nor Partigen plates by Behring. The results were as follows: mean AAT concentration in the subretinal fluid of all the patients operated on for RRD was 0.739 +/- 0.475 g/l and CER was 0.186 +/- 0.122 g/l. Mean AAT concentrations did not differ statistically significantly between the group with bigger and smaller retinal detachments while CER level was higher in the group with a bigger RRD. Mean AAT and the CER concentration was statistically significantly higher in the group with RRD lasting from 1 to 3 months compared with the group with RRD lasting shorter than one month. CONCLUSIONS: the fact of finding acute phase proteins in subretinal fluid suggests that they may play a part in the pathogenesis of RRD by affecting the intensity of local inflammatory reaction. PMID- 12898926 TI - Does a correlation between transaminase activity and the size of the liver in psoriatic children exist? AB - Psoriasis is a systemic disease. On the basis of the literature it has been stated that there is a possibility that a disease which is going on in the skin may be reflected by the increase in the activity of the hepatic cell. This activity was estimated by the McLaghan units, concentration of aspartate and alanine aminotransferases and morphometric measurements of the liver in 3 oblong dimensions. The study was conducted on 56 children suffering from psoriasis and 32 healthy ones matched with sex and age. None of the investigated parameters differs statistically between ill and healthy children. No association was stated between psoriasis and the enlargement of the liver dimensions by means of ultrasonography or the McLaghan units, ALAT and ASPAT. PMID- 12898927 TI - The effect of epilepsy on emotional state in Hamilton and Beck questionnaire studies. AB - Patients with epilepsy present higher rate of psychological and psychiatric problems and emotional disturbances are estimated as the most frequent ones with prevalence of depression about tree times higher than it was observed in general population. In the presented study the occurrence of depressive symptoms was compared in a total of 60 adult patients with newly recognized and chronic epilepsy during remission of illness as well as pharmacoresistant. 21-items Hamilton Depressive Scale and Beck Depressive Inventory were applied once in each testified patient, consecutively by a physician or a patient himself during interictal period. Our study has found a higher level of depressive signs in newly diagnosed than in chronic patients during remission of illness but also in patients with an active process. The scoring rates both on HDRS and BDI were respectively characteristic of moderate or mild intensity of depressive disturbances. This comparison allows us to make a suggestion that diagnosis of epilepsy with its well-known stigmatization and limitations of different life activities may act as a stressing factor that enhances anxiety and depressive reactions in early phase of illness. PMID- 12898928 TI - Effectiveness of adjunctive therapy with tiagabine in mentally disabled patients with epilepsy. AB - Previous reports justified positive impact of tiagabine, a new-generation antiepileptic drug on neuropsychological functioning as well as minimal risk of psychiatric exacerbations. In this prospective, open, observational, non comparative, short-term--6 months' study we evaluated efficacy and tolerability of tiagabine as add-on in polytherapy in 30 young adults with both refractory epilepsy and mild or moderate mental retardation. About 40% of patients experienced improvement in seizure frequency of 50% or more and none complained of seizure deterioration. Overall seizure frequency fell down from a mean 9.9 at the baseline to 6.3 seizure days per month. The seizure rate was reduced from 14.9 to 9.2 per month after 24 weeks of add-on phase. Adjunctive tiagabine therapy appeared to be associated with reduction in overall seizure severity characteristics expressed as enhancement of mild seizures from 30 to 50% of patients and reduction in severe seizures from 30 to 16%. Responders reported an improvement in most health-related quality of life domains but mostly cognition, medication effects and social functioning. The majority (90%) of patients did not report additional adverse effects with tiagabine; the most common complains associated with central nervous system (vertigo, weakness, nervousness) were transient and did not cause discontinuation. Blood counts and liver functional tests did not show any clinically relevant changes. In this short-term observation tiagabine seemed to be a beneficial antiepileptic drug for mentally retarded patients with epilepsy as it decreased seizure frequency and severity, improved the patients' sense of the quality of life without enhancement of the risk of adverse effects in polytherapy. PMID- 12898929 TI - The level of total sialic acid, alfa-antitrypsin, ceruloplasmin in the serum of patients with choroidal melanoma. AB - Melanoma of the choroid is a fatal disease because of its metastases. The quest is ongoing for more reliable serum markers for detecting and staging ocular melanoma. Total serum sialic acid and acute phase proteins are valuable adjuncts in the management of malignancies, including melanoma. The aim of the paper was to asses the level of total sialic acid (TSA), total sialic acid to total protein (TP) ratio (TSA/TP) and the level of alfa-antitrypsin (AAT) and ceruloplasmin (CER) in patients with choroidal melanoma. The concentrations of TSA, TP, AAT and CER were evaluated in 61 patients with choroidal melanoma and 84 healthy controls. 36 patients had larger tumors and 25 patients had smaller melanomas. 36 patients were treated with brachytherapy. The mean concentration of TSA in all intraocular melanoma patients was 84.86 +/- 19.37 mg/dl and was significantly higher than in control group 53.63 +/- 8.47 mg/dl (p < 0.001). TSA level was significantly higher in patients with large tumors than in those with smaller choroidal melanomas. There were not differences between groups of patients treated with brachy-therapy and those not treated. TSA/TP in melanoma patients was 11.88 +/- 2.97 and it was higher than in control group 7.32 +/- 1.21 (p < 0.001). AAT level was 236.56 +/- 141.53 mg/dl in the group of melanoma patients and in the control group was 226.42 +/- 46.74 mg/dl but the differences were not statistically significant. The concentration of CER in the study group was 28.25 +/- 11.01 mg/dl and in the control group it was 29.56 + 6.33 mg/dl but the differences were not statistically significant. The assessment of TSA in blood serum may be useful in evaluation of patients with choroidal melanoma. PMID- 12898930 TI - Influence of adenosine receptors on the development of caerulein-induced acute pancreatitis. AB - Acute pancreatitis leads to hypoxia caused by vasoconstriction and to activation of lysosomal and digestive enzymes resulting in pancreas autodigestion and damage. This causes activation of leucocytes and increased expression of adhesive molecules enabling margination and adhesion of activated leucocytes to the endothelium. Activated leucocytes are the source of proinflammatory cytokins and oxygen-free radicals which intensify the inflammatory response. The reports indicating that adenosine may prevent activation of the above-mentioned processes in ischaemia prompted us to undertake this study. The study was performed in two stages. The first stage was to evaluate the effects of agonists and antagonists of adenosine receptors on normal pancreas while the second one was to determine the influence of these substances on the development of caerulein-induced acute pancreatitis. During the first stage, the animals were injected intraperitoneally with the substances examined: the A1 receptor antagonist--DPCPX, the A2 receptor agonist--CGS 21680, the A2 receptor antagonist--ZM 241385 and the A3 receptor agonist--IB-MECA and then received intravenous saline. The control animals were subjected only to the 12 h intravenous infusion of 0.15 M NaCl. During the second stage, after the intraperitoneal administration of adenosine receptor agonists and antagonists (as in the first stage), acute pancreatitis was induced with the 12 h intravenous infusion of 5 micrograms/kg/h caerulein. Identical acute pancreatitis was induced in the control animals, however no other substances were administered. The pancreatic tissue samples were collected directly after intravenous infusion. The severity of inflammatory processes in the pancreas was evaluated on the basis of the plasma amylase activity, pancreatic weight and enhancement of histopathological changes observed in this organ. In the animals infused with saline alone, no effects of the substances examined on the pancreatic weight, plasma amylase activity and histopathological features were observed. The intravenous caerulein infusion induced acute pancreatitis expressed as bigger pancreatic weight, increased plasma amylase activity and tissue damage (oedema, cell vacuolization, leucocyte infiltration). The A2 receptor agonist administration preceding the induction of acute pancreatitis decreased the pancreas damage caused by caerulein. Lower weight of the pancreas and decreased plasma amylase activities were observed; on histopathological examination- oedema, leucocyte infiltration and intensity of alveolar cell vacuolization were lower. On the other hand, intraperitoneal pretreatment with the A2 receptor antagonist intensified the pancreas injury. The A1 receptor blockade and A3 receptor stimulation in the animals injected with caerulein did not affect the pancreatic weight, plasma amylase activity or histopathological picture of the organ. The administration of A2 receptor agonists decreases the organ injury in caerulein-induced acute pancreatitis in rats. This action may result from modulating effects of these substances on different stages of the cascade of inflammatory reactions. However, the present study did not reveal any effect of the A1 receptor agonist or A3 receptor antagonist on inflammatory processes in the experimental model described. PMID- 12898931 TI - Ways of spending leisure time by the third year-students of the Faculty of Pharmacy, Medical University of Lublin. AB - The aim of the study was the assessment of leisure time among third-year students from the Faculty of Pharmacy of the Medical University of Lublin. It analysed quantity of time devoted to school activity and ways of spending free time. The study involved 114 students (82 women and 32 men). The study revealed that women had less free time than men, who, most probably did not attend some lectures. The most popular activities among the questioned students were: sleeping (average 6.8 hours a day), studying (average 3.6 hours a day), listening to the radio (average 2.9 hours a day), talking with friends (average 1.9 hours a day), personal hygiene (average 1.1 hours a day), watching TV (average 1.1 hours a day), housework. Students devoted the least of their free time on active rest, for example walking (women did it more often than men) or practising sport (more popular among men). Cultural life of the students consisted only of meetings with friends and going to the cinema (women did it more often). The least popular way of spending free time was going to the theatre, opera, concerts and exhibitions. Few students spent their time working. Their number increased significantly during holidays. The way of spending free time by third-year students from the Faculty of Pharmacy (both men and women) during the day was similar, differences related only to the amount of time devoted to each activity. PMID- 12898932 TI - The influence of diazepam on the behaviour of rabbits in spontaneous conditions. AB - The study dealt with a popular drug diazepam also known as Relanium. Two doses of the substance were tested: 0.2 mg/kg and 1 mg/kg. The experiment was carried out on 10-rabbit groups for two hours. The animals were observed in peaceful environment of the laboratory after one hour of adaptation. There were distinguished a few phases of behaviour: active and passive observation, comfort, tension, grooming, food and water intake. Both doses eliminated completely the tension phase. Changes in other phases were similar to each other but depending on the dose there were differences in strength and duration. The smaller dose caused significant prolongation of the comfort phase at the expense of active and passive observation time decrease during the first hour of observation. The second hour trends were not changed but lost their statistical relevance. The changes caused by the bigger dose were almost identical and they were statistically significant during both hours of the experiment, but their amplitude was stronger than that caused by the lower dose. The outcome of the experiment confirms anxiolytic, sedative and miorelaxant properties of diazepam. PMID- 12898933 TI - The influence of Atorvastatin on ultrastructure of rat pancreatic exocrine cells in the course of experimental intoxication with ethanol. AB - The experiment was carried out on mature Wistar male rats. The animals from experimental group I received Atorvastatin in the dose 0.28 mg/24 h for 3 weeks. Experimental group II received 20% ethanol ad libitum apart from Atorvastatin. Ultrastructural examinations of pancreatic exocrine cells showed the stimulation of secretory processes in cells of animals receiving Atorvastatin and considerable damage of cell organelles in experimental group II, more intense than in the pancreas of control animals which drank ethanol. PMID- 12898934 TI - Intravascular thrombosis as a result of central venous access. AB - Central venous access represents one of the most basic therapeutic procedures in modern medicine. Unfortunately, numerous advantages that result from maintaining a central venous line are accompanied by some complications among which the venous thrombosis is the most significant clinically. The study was designed to assess frequency and natural history of this complication in the setting at a multi profile clinical hospital. Central venous cannulation was performed by a fully qualified anaesthesiologist in every case. There were 887 cannulations and only 5 patients with clinically significant venous thrombosis. The analysis of the collected data allowed us to state that the frequency of intravascular thrombosis is low, but this complication is often associated with extensive impairment of patency of the central veins. Full recanalization is not always achieved regardless of the treatment applied. Pulmonary embolism in the course of central venous thrombosis was diagnosed in one patient only and appeared as a multiple and fine X-ray infiltrates. It seems that in the presence of permanent or even life threatening complications of central venous thrombosis their risk should be minimized by frequent examination of the cannulation site and early initiation of antithrombotic treatment. PMID- 12898935 TI - Changes in health status of the Polish population after 1990. AB - The health status of the Polish population in the past decade has been strongly influenced by economic and political transformations. One of the measures of such transformations is the analysis of alteration in the structure of causes of deaths and mortality rates. In order to present the changes in health status of Polish population that occurred after the year 1990, in the paper there have been analysed epidemiological data in the form of mortality rates and the causes of deaths in the Polish population in the years 1990-2000. PMID- 12898936 TI - Evaluation of state of deciduous dentition in children in Lublin day nurseries. AB - 408 children aged 1-4 years from Lublin nurseries were examined. The paper aimed at evaluation of the mean number of dmf and the level of oral hygiene in these children. The studies confirmed high caries frequency as well as high OHI index. PMID- 12898937 TI - The role of oral hygiene in dental caries prevention in children and adolescents. AB - The role of oral hygiene in prevention of caries and periodontal diseases has been presented based on the literature and own clinical experience. PMID- 12898938 TI - High resolution computed tomography (HRCT) in diffuse pulmonary intraparenchymatous disorders. AB - The value of high resolution computed tomography (HRCT) was presented in the material of 62 diffuse pulmonary intraparenchymatous disorders. Visualisation of milk-glass type opacities, honeycomb picture, nodular changes, alveolar emphysema, usually associated with intralobular or periseptal, was analyzed. Expirational HRCT section facilitate the discovery of air traps. HRCT make possible differentiation of active changes from irreversibles (fibrous). PMID- 12898939 TI - Insulin resistance, parameters of carbohydrate metabolism, anthropometric parameters, interactions in diabetic patients. AB - The aim of the study was to investigate relationships between insulin resistance index and basic parameters of carbohydrate metabolism and anthropometric values. We investigated patients with diabetes mellitus t.2 (n = 36) treated with oral hypoglycemic agents. The control group represented patients with normal glucose tolerance (n = 37). Both, the group with diabetes and control had similar values of anthropometric parameters. The insulin resistance index showed significant correlations with fasting plasma insulin and glucose concentrations and HbA1C level in both groups. In the group with normal glucose tolerance we showed significant correlations between insulin resistance index and anthropometric parameters (waist circumference, BMI, body weight, WHR). We did not find any significant relationships between insulin resistance and anthropometric parameters in the diabetic group. PMID- 12898940 TI - The loss of alveolar processus, mandible density and bone tissue structure in reference to the computerized analysis of intraoral radiography. AB - 98 patients treated in the Department of Dental and Maxillofacial Surgery, Medical University and Institute of Rural Medicine in Lublin had radiovisiographically tested premolar teeth area on the left side. Reviewing radiovisiographical picture, there were defined bone mineral density and mandible bone structure and degree of loss of alveolar processus. It was concluded that the degree of loss of alveolar processus is significantly correlated with the researched patients' age, and is inversely correlated with mineral density and equivalent of mandible bone mineral contents. The achieved results prove that demineralisation changes can significantly influence teeth loss. PMID- 12898941 TI - Assessment of density and mandible bone structure in patients with generic osteoporosis symptoms. AB - In a group of 130 patients with diagnosed osteoporosis and in 110 patients in control group radiovisiographic test of mandible bone structure had been carried out. Digital images analysis was carried out using Software Radiograph Workshop. It proved that in the group of patients with osteoporosis process in comparison with the control group there has been observed a significant deterioration of mandible bone density, defined as reduced bone mineral contents (BMC), reduced bone mineral density (BMC), and reduced mandible bone density (BD). Bone structure analysis showed in the control group of the treated patients with osteoporosis lower than in the control group quantity of structure of trabecular number (STN), deterioration of bone trabecular density expressed in % of area and greater structure of trabecular width than that in healthy patients. PMID- 12898942 TI - Effect of age and gender on the condition of bone tissue of stomatognathic system. AB - The 240 patients, with the mean age 50.3 year for males and 54.4 years for females treated in the Department of Dental and Maxillofacial Surgery Medical University and Institute of Rural Medicine in Lublin had radiovisiographically tested premolar teeth area on the left hand body side. Digital radiovisiographical picture was developed using computer software Radiograph Workshop, and obtained factors of bone mineral density and mandible bone structure were reviewed with regard to gender and age of the treated. It was concluded that relative density, mineral density and bone mineral contents is significantly greater with male than female patients, and is deteriorating significantly with age. Age and gender of the treated in a lesser degree influences on radiological structure factors resulting from radiovisiographical picture analysis using Radiograph Workshop. PMID- 12898943 TI - Influence of estrogen deficiency on the copper level in rat teeth and mandible. AB - On the basis of atomic absorption spectrophotometry rat teeth and mandible copper levels in experimental postmenopausal osteoporosis and after administration of 17 beta-estradiol were measured. The results showed that estrogen deficiency leads to decrease in copper content in rat teeth and mandible, and giving of 17 beta estradiol positively influence the content of mineral components in the examined tissues. PMID- 12898944 TI - Results of Nd:YAG laser capsulotomy in posterior capsule opacification. AB - The most common postoperative complication of cataract extraction is posterior capsule opacification. It causes the deterioration of visual acuity, however, there is no effective method to prevent it. The posterior capsule opacification is treated by surgical or laser capsulotomy. The authors analysed the results of Nd:YAG application in discission of the opacified posterior capsule. Twenty-eight consecutive cases (23 patients) of extracapsular cataract extraction and phacoemulsification with implantation of posterior chamber intraocular lenses were reviewed. In all cases of capsulotomy the Nd:YAG laser of Q-switch Zeiss type was applied. The improvement of visual acuity was achieved in 25 cases (89%). The best visual acuity (0.8-1.0) was obtained in 72% of eyes and in 22% the improvement was from 0.25 to 0.5. In two cases visual acuity after capsulotomy deteriorated because of concomitant glaucoma. In one case repeated capsulotomy did not turn out to be successful. It was caused by diabetic neuropathy of the optic nerve. We did not observe any serious complications following Nd:YAG laser capsulotomy described by the others: the intraocular pressure elevation, cystoid macular edema, intraocular lens injury, retinal detachment, iridocyclitis. Nd:YAG laser capsulotomy is one of the best methods of treatment of posterior capsule opacification. It appears to be effective, safe, non-invasive and it does not require patient hospitalisation. PMID- 12898945 TI - Application of scintigraphy MDP 99 mTc in the diagnosis of osteoarticular pains of obscure etiology. AB - The three-phase scintigraphy of bones is useful in detecting disorders of bone tissue perfusion metabolism in patients with the occult etiology of joint pains. The bone scintigraphy with application of MDP plays the main role in the orthopaedic diagnosis. It is of the greatest importance when radiograms are not normal. In spite of application of the other modern methods it is the technique of choice in the early diagnosis. PMID- 12898946 TI - Urinary iodine excretion in inhabitants of rural region living in the Lublin Province. AB - Iodine is the element necessary for the human organism to function properly. Iodine is essential especially for the synthesis of thyroid hormones that regulate the cellular metabolism of the whole organism. The purpose of the work was to estimate the concentration of iodine in urine among the inhabitants of a typical agricultural region as a simple method of detecting deficiency of iodine in the human organism. The biggest iodine excretion was observed in a group above 40 years of age. The amount of the excretion of iodine together with urine was similar among men and women up to 40 years old. Monitoring of iodine concentration in urine is a sensitive marker of iodine deficiency in the human organism. PMID- 12898947 TI - Leisure time activities in teenagers in urban and rural areas. AB - The work aimed to determine the leisure time activities in teenagers on weekdays, weekends, during winter and summer holidays. Vast majority of teenagers spend their leisure time resting in a passive way, i.e. watching TV or playing computer games irrespectively of the season. As a result of this, the number of kids with posture defects increase. On weekdays the country teenagers spend much more time doing outdoor sports and games than town children. They also more often help their parents and less frequently travel away from their homes than children living in towns. PMID- 12898948 TI - Results of coronary arteries bypass grafting in patients over 70 years old. AB - At present, according to the increase of average age in modern societies, the patients who are over 70 years old and require coronary arteries reconstruction constitute a constantly growing group. In the Cardiosurgical Clinic of the Medical University of Lublin 30 coronary arteries bypass grafting operations in over 70-year-old patients were performed in 2000. The operations were performed using extra-body circulation. In the early postoperation period 3 patients died (10%), 2 patients (6%) required applying mechanical support of the left ventricle (IABP). 2 patients (6%) needed reoperation because of excessive drainage in the immediate postoperational period. CABG in patients over 70 years of age are connected with higher "perioperational" risk, prolonged recovering period and higher mortality. According to the results of the survey, the overwhelming majority of patients at that age gets improvement in their health and in the quality of life. PMID- 12898949 TI - Mercurialism in the material of the Regional Centre of Acute Poisonings of the Institute of Rural Medicine in Lublin. AB - Against the background of clinical symptoms of mercury poisonings a case of suicidal poisoning which had favourable course and prognosis was presented. The impact of toxic renal damage and different CNS symptoms was emphasised. PMID- 12898950 TI - Arteriovenous angiomas in pictures of digital subtractional angiography (DSA). AB - The value of digital subtractional angiography (DSA) was presented in the material of 16 arteriovenous angiomas found in the group of 591 angiographies of cerebral arteries. Visualisation of arterial supplying vessels, vascular mass, nucleus and draining vessels was analyzed. Angiograms were correlated with CT contrast sections to reconstruct coexisting intracerebral and subarachnoid hemorrhages. The degree of visualization of mass effect in the two examinations was compared. There was emphasized the role of angiographic demonstration of the sucking effect of angiomas in the so-called stealing syndromes. PMID- 12898951 TI - The activity of dental caries in students of the Faculty of Dentistry, the study with the use of microbiological and biochemical tests (Dentocult SM). AB - The aim of the paper is to answer the question whether the Dentocult SM tests could be used to study dental caries activity in the 20-25 age group. 66 students of the Faculty of Dentistry, Medical University of Lublin, were included in the study. In all cases dental state was assessed by using average numbers of DMF and DMFs Ds. In the microbiological examinations a ready-made medium from Dentocult SM set was used. The results revealed that the risk of dental caries in the population studied cannot be determined only on the basis of Streptococcus mutans values. PMID- 12898952 TI - General practitioners' opinions on the intake of painkillers among patients. AB - The goal of this paper is to evaluate the consumption of painkillers among patients on the basis of the opinions of general practitioners. The most frequent ailments including pain symptoms were the cases of long-continued pain (64.3%); less frequent were acute pain syndromes in the course of a disease (35.7%). The phenomenon of the excessive use of painkillers among patients with long-continued pain syndromes is observed by the GPs. Uncontrolled self-treatment is possible owing to an easy access to this type of medicaments. The excessive use of analgesic medicines in therapy frequently results from the lack of simultaneous application of other methods of pain treatment e.g., in physiotherapy, psychotherapy. Long lasting use of various types of painkillers can lead to drug addiction. This problem is observed by over a half of the GPs (67.1%). PMID- 12898953 TI - Long-term survival after bronchial sleeve resections. AB - Between February 1990 and February 2000, 53 anatomical pulmonary tissue resections with sleeve bronchial resections have been performed in the Department of Thoracic Surgery of Medical University of Lublin. The aim of the present study was the evaluation of long-term results of operative treatment. Most of the cases (86.8%) were patients with non-small cell lung cancer. Three perioperative deaths (5.7%) were recorded. The mean time of survival for all patients was 45.6 months and for NSCLC patients was 41.2 months. The cumulative 5 years' survival rate in cancer patients was 38.6%. Preoperative performance status, presence of N2 and non-radical resection have been recognised as significant prognostic factors in cancer patients. In our opinion, in view of comparable efficacy, low morbidity, mortality and functional benefits pulmonary sleeve resections should be considered in every patient with favourable anatomical and functional conditions. PMID- 12898954 TI - Causes and circumstances of injuries in children. AB - The problem of the incidence of traumas among children is still topical and statistical data confirm an increasing tendency in the last decades. The purpose of the work was to estimate the causes and circumstances of injuries in children at the age of 7 to 15, who were admitted to Surgical Ward of Specialist Paediatric Hospital in Kielce between 30th June 1997 and 30th June 1998. A retrospective analysis of medical documentation concerned 768 patients. Our studies showed that the biggest number of injuries was related to children 10-12 years old (46%). The majority of patients were boys. Most injuries occurred in summer (33%). In the studied population dominant were accidents at home (56%) and then road accidents (18%), injuries at school (13%) and on farms (7%). Accidents and injuries occurred more often in children from the rural areas (53%). PMID- 12898955 TI - Life style and the risk of development of circulatory system diseases. AB - Circulatory system diseases (c.s.d.) belong to the group of civilization diseases. The risk factors of circulatory system diseases are: stress, overuse of alcohol, smoking cigarettes, bad eating habits, sitting life style. Calorie overconsumption, animal fat rich diet, sitting life style result in the development of android obesity, hypercholesterolemia, that enhance atheromatosis. The most dangerous consequences of atheromatosis are: angina pectoris, hypertension, myocardial infarction, brain insult, type II diabetes. The aim of the work was to evaluate life style and its correlation with development of circulatory system diseases. The results obtained significantly indicate how important is health education about the danger of widely understood unhealthy life style. Propagation of staying healthy model with emphasizing real profit resulting from leading healthy life style is the only righteous way to improve it. PMID- 12898956 TI - Starting with the hospice/palliative care project--the philosophy and the practice. AB - The goals of the palliative care is the maximizing of the quality of life of people with the chronic progressive illness, their families and those who care about them. The palliative care is characterized by the interdisciplinary and patient-centered approach which consists of five dimensions: the sphere of the biological, psycho-social and spiritual functioning centered on the patients life, and the others: the care for the family and care for ourselves concerned with the care for the caregivers. The authors describe the development of the palliative care in the last twenty years and stress the importance of the difficult but necessary balance-keeping between the philosophy, mission and the practical needs. PMID- 12898957 TI - Palliative care professional education in the new millennium: global perspectives -universal needs. AB - In the 20th century, the palliative care is based mostly upon the Anglo-Saxon and Christian contemporary model of care. Is such a model sufficient and could be transformed to the places outside of Judeo-Christian based, west civilization? Is such a model transcultural and could be used for problems solving for those who are professionally involved in the end-of-life care? The universal needs of the dying human being and the family are seen by the authors in the global perspective, and that perspective is tailored as a base for the transcultural model of the education for the professionals within the palliative care. PMID- 12898958 TI - Surgical treatment in Paget's disease of the breast. AB - The extent of the operation in Paget's disease of the breast depends largely on the degree of clinical advancement of neoplasm as well as counter-indications for breast conserving therapy. Despite the increasing use of breast conserving therapy, rather few such operations of this kind have been performed in our department. We have carried out 29 mastectomies and only 6 breast conserving operations. No matter what type of therapy was applied the percentages of 5-year survivals for the patients with non-invasive component and for the ones with invasive component were respectively 89.76% and 49.32%. PMID- 12898959 TI - The possibilities of surgical cover of skin defect in locally advanced breast cancer. AB - Three types of surgical procedure were applied to cover skin defect in locally advanced breast cancer after simple mastectomy. The skin defect was impossible for primary suture. In four cases we used latissimus dorsi myocutaneous flap with a very good effect. In two cases skin flap from the abdomen was used, and one case skin graft. The female patients survived 2.1 years with better quality of life. PMID- 12898960 TI - Ultrastructural changes of pancreas of white rats after experimental administration of ethanol and cephalexin. AB - The experiment was carried out on Wistar rat males weighting about 200 g. Animals from experimental group I received 20% ethanol for drinking (ad libitum), animals from experimental group II--cephalexin in the dose of 42 mg/24 h, animals from experimental group III--cephalexin and ethanol in the mentioned doses. After 10 days the animals were decapitated and pancreases were collected for ultrastructural examinations in electron microscope. The performed experiments showed that 10-day administration of ethanol causes mainly the decrease of amount of ribosomes and zymogen granules in pancreatic exocrine cells, whereas cephalexin causes increase of amount of these organelles. Administration of both ethanol and cephalexin causes reversible degenerative changes including distinct decreasing of the number of ribosomes, swelling of many mitochondria and the presence of myelinic structures inside cells. PMID- 12898961 TI - Degenerative changes of the vertebral column in spatial imaging of 3D computed tomography. AB - In a group of 38 patients with radicular pain syndromes diagnostic value of spatial reconstructions with computed tomography (3D CT) was assessed in examinations of bone structures of the vertebral column. It was found that 3D CT is a technique of choice in the assessment of degenerative stenosis of the vertebral canal, internal surface of the vertebral canal, bone narrowings of intervertebral foramens and lateral recesses. PMID- 12898962 TI - Selected determinants of the quality of hospital care. I. Accreditation and effectiveness of managing a therapeutic team. AB - In order to provide a higher level of the quality of medical services an external process of evaluation of health care units called accreditation has been implemented. Obtaining the Quality Certificate by a hospital greatly depends on knowledge and engagement of the staff in the accreditation process as well as on the effective managing of therapeutic teams by ward head/managers of clinics. Therefore, studies were undertaken to discover the knowledge of requirements concerning the possession of the Quality Certificate and how managing therapeutic teams is evaluated by physicians and nurses from hospitals with and without accreditation. The study covered 560 physicians and nurses from 4 hospitals with accreditation and 17 hospitals which did not possess the Quality Certificate. The research method was a diagnostic survey, and the technique--a questionnaire form. The material was subject to statistical analysis. Significant differences were observed in the knowledge of requirements concerning the possessing of the Quality Certificate between the staff of hospitals with accreditation, compared to the respondents from health units without the Quality Certificate. Respondents from hospitals without accreditation evaluated the effectiveness of managing therapeutic teams by ward head/managers of clinics in more negative terms, compared to the staff of hospitals which possessed the Quality Certificate. PMID- 12898963 TI - Selected determinants of the quality of hospital care. II. Managing a therapeutic team. AB - The effects of teamwork depend to a large extent on the organizational skills of the manager. In health care units a physician/ward head is responsible for coordinating the work of a therapeutic team. The study was undertaken to discover to what extent doctors and nurses are aware that the ward head manages the work of the therapeutic team, and how they evaluate the skills of their managers. The study covered 161 doctors and 339 nurses from 4 hospitals with accreditation and 17 health units which did not possess the Quality Certificate. The study was conducted by the method of a diagnostic survey, and the technique was a questionnaire form. The results of the survey showed that charge nurses in hospitals with accreditation and ward heads in hospitals without the Quality Certificate significantly more often perceived the effectiveness of managing a therapeutic team by a ward head/manager of a clinic in positive terms. A greater number of negative evaluations were expressed by charge nurses in hospitals without accreditation and ward head nurses in hospitals with the Quality Certificate. It was confirmed that doctors and nurses from hospitals with accreditation significantly more frequently perceived the ward head as the manager of the therapeutic team, compared to the staff of hospitals without the Quality Certificate. PMID- 12898964 TI - Selected determinants of the quality of hospital care. III. Scope of duties, authorization and responsibility among members of a therapeutic team. AB - An important precondition for an effectively working team is the distribution of competence among the members of the team. The accreditation criteria for Polish hospitals lay down a requirement concerning the formulation in a written form of the scope of duties, authorization and responsibility at individual workplaces. This conditions a high level of services provided by the members of a therapeutic team. Therefore, the study was undertaken in order to discover whether there was an explicit share of competence and if it exerted an influence on the level of performance of the scope of duties, authorization and responsibility among members of therapeutic teams in hospital wards. The study covered 55 ward head/managers of clinics, 72 ward head nurses, 106 assistant doctors and 327 charge nurses from 21 hospitals, who were employed in 4 hospitals which possessed the Quality Certificate, and in 17 hospitals without accreditation. The study was conducted by the method of a diagnostic survey, the technique being a questionnaire form. The results of comparative analysis showed that among members of therapeutic teams in hospitals with the Quality Certificate the distribution of occupational competence was significantly more often observed. The greatest number of respondents in this group possessed the scope of duties, authorization and responsibility in a written form, compared to physicians and nurses from hospitals without accreditation. Moreover, it was confirmed that the most frequent lack of the scope of duties in a written form as well as the greatest discrepancies concerning the performance of occupational tasks were noted among the members of therapeutic teams in clinical hospitals without accreditation. PMID- 12898965 TI - Fatal intracranial hemorrhage after double valve replacement in infective endocarditis. AB - Endocarditis is a life-threatening disease, although it is relatively uncommon. The complications of infective endocarditis may involve any organ system. Cardiac and extracardiac complications including neurologic, vascular and renal diseases are common and are usually caused by either embolization of vegetation or immune complexes. This case report presents a 31-year-old male with combined mitro aortic valves defect and infective endocarditis on mitral valve. The replacement of mitral and aortic valves prosthesis were performed. During postoperative period low output heart failure, renal failure and respiratory failure developed. 25 days after the first operation the reoperation was performed because of mitral prosthesis insufficiency and thrombus just under the valve. On 31st day after the first operation patient died because of subdural hematoma and acute cerebral oedema and encephalomalatia. PMID- 12898966 TI - Benign prostatic hyperplasia, prostate cancer and other prostate diseases diagnosed as a result of screening procedure among 1,004 men in the Lublin district. AB - Urological problems especially connected with prostate diseases appear in older men: prostatitis, benign prostatic hyperplasia, prostate cancer and others. Prostate cancer is the most common cancer in men and the second leading cause of deaths in male population. Therefore, it seems important to early detect prostate cancer in general practice setting due to screening procedures such as digital rectal exam and prostate-specific-antigen test. The aim of the study was to examine the character of diagnoses of a prostate disease among 1,004 men of the Lublin district who reported to a doctor during screening procedure carried out in urology outpatient clinic at Clinical Hospital in Lublin in the year 2000. After physical examination urologists initially diagnosed a prostate disease and sent men suspected to have a prostate cancer to further investigations. There was studied age and place of living. Benign prostate hyperplasia was the most common diagnosis made in 77.1% of subjects. It occurred most often in men aged 51-70 years. Prostate cancer was suspected in 3.5% of subjects. Frequency of benign prostate hyperplasia and prostate cancer suspicion increased with age. On the basis of studies screening procedures seem beneficial in the early detection of prostate cancer in men over 50. PMID- 12898967 TI - Objective assessment of self-care and non-professional care: a proposal. AB - Nursing care today, both in theory and practice, consists not only of professional care rendered by nurse but, to a significant degree, involves also self-care and non-professional care. This tendency is also reflected in the philosophical premises of Polish Nursing. The interest in self-care and non professional care in Poland, and, consequently, in the D. Orem's self-care deficit theory of nursing, as well as attempts at their implementation in nursing practice inspired the present attempt to develop a tool for objective assessment of these two types of care. The proposed Scale of Patient's and Care Provider's Agency (SPCPA) allows for the objective assessment of self-care as patient's self care agency, and of non-professional care as the main care provider's agency measured in points. The aim of this study is to describe the tool and to assess its two psychometric properties: reliability and validity. The results of the statistical testing of the SPCPA show that the tool is capable of assessing precisely both the patient's self care agency and of the main provider's caring agency. PMID- 12898968 TI - Caring in nursing--conditioning of realisation. AB - Caring manifested by the concern of a nurse, attention and carefulness of activities with respect to a human being in need is the essence of nursing care, while caring actions constitute the most independent scope of nurses' activities. The aim of the study was to determine whether the nurses in their everyday occupational practice understand the core of caring actions and perceive the conditioning of their realisation. The study covered 130 nurses employed in inpatient health facilities. The survey was conducted by means of a questionnaire form containing variously constructed questions. The results of the study showed that nurses undertook caring actions 'willingly' more often than 'very willingly', the percentage of respondents undertaking caring actions in category 'very willingly' increasing with the duration of employment. Over 86% of nurses in the study independently undertook caring actions, as soon as they recognized patients' demand; however, only 7% of respondents admitted that caring actions determine the distinct character of the nursing profession. According to the greatest number of respondents the following factors determined the quality of caring services: organization of work in the ward, provision of material and means for nursing, personality-related factors, incentives for work, and supervisor's requirements. Nurses applying the method of individual work with a patient undertook caring actions more often than those working by the method of traditional nursing. PMID- 12898969 TI - Allergy to house dust mites in primary health care subjects with chronic or recurrent inflammatory states of respiratory system. AB - Chronic and recurrent respiratory tract disorders are a frequent problem in general practice. The purpose of the study was to investigate the role of hypersensitivity to house dust mites in respiratory tract diseases in general practice patients. We tried to assess the influence of determined risk factors exposure on development of respiratory tract allergy. Patients from family practitioners surgeries with chronic or recurrent respiratory tract symptoms who had no diagnosis of allergy were recruited to the study (n = 89). All patients responded to a questionnaire focused on history of symptoms, atopic conditions in family and exposure to determined environmental factors like dwelling conditions, obstetrician history, diet in the first year of life. All patients underwent skin prick test with common inhalant allergens. Families of the patients were asked to participate in the study. Families who agreed to take part also responded to the questionnaire and underwent skin tests. In patients and their families blood samples were taken to determine total IgE and specific IgE antibodies to mites allergens. Dust samples were collected by vacuuming of patients' bedroom carpets and mattresses to determine house dust mites allergens concentration. Data on 30 complete patients family sets of their brotherhood, mother and father were collected. Total and specific serum IgE antibodies were determined by disc enzyme immunoassay (Analco). Mites allergens concentration in dust was measured by simple Acarex strip test (Nexter). The results of the assays (positive skin tests and/or elevated levels of specific IgE) showed allergy to house dust mites in 24 of 89 study patients from general practitioners surgeries (27%). The prevalence of chronic rhinitis, recurrent bronchitis, chronic or recurrent cough, wheezing, dyspnoea was higher in allergic than in nonallergic subjects. Patients with the diagnosis of allergy to house dust mites had usually worse dwelling conditions. Especially the influence of dampness in flats on several respiratory symptoms was observed. Subjects who had been found to be allergic were more frequently exposed to higher concentrations of house dust mites allergens in bedroom carpets and in mattresses. In allergic children early introducing of sensitizing components into the diet in infancy related to shorter breast feeding was observed. PMID- 12898970 TI - Sleeping disturbances and pharmacological/nonpharmacological interventions in old people. AB - Sleeping disturbance constitutes a major problem occurring in 25% of people aged over 65. Among the most frequent reasons of insomnia one can mention the overuse of coffee, alcohol, smoking, using some drugs, stress, depression, neurosis, dementia, chronic diseases. Idiopathic insomnia constitutes less than 5% of cases. Before the treatment of insomnia is started, its cause should be found and it should be determined if it is transient or chronic. The treatment of insomnia can be based on pharmacotherapy and non-pharmacotherapy methods. Behavioural therapy is mentioned among non-pharmacotherapeutic methods. It aims at changing the sleep-related habits. Pharmacotherapeutic treatment (still mostly based on benzodiazepines) should not last longer than 2-3 weeks and drugs of short elimination T 1/2 (below 6 hours) should be used. Side-effects of this group of drugs should always be taken into consideration. It should be remembered that sleep disturbances can be symptoms of other diseases and their treatment cannot consist in prescribing hypnotics only. PMID- 12898971 TI - Primary health care and geriatric medicine in Denmark. AB - In Denmark with its population of 5.1 million, 15% of inhabitants are people aged over 65 and--just like in other European countries--the percentage of elderly people has increased in the recent years. A model of geriatric care in Denmark was presented in this study. The Danish health care system has been briefly described in respect of the care for the elderly. Life conditions of the elderly people in Denmark as well as social and health care services provided for them were discussed. PMID- 12898972 TI - Diagnosis, treatment and registration of urinary tract infections in geriatric patients. AB - Bacteriuria (> or = 10(5) CFU/ml) is a very common phenomenon in elderly people, occurring twice as frequently in women than in men. There are symptomatic and asymptomatic types of bacteriuria. Risk factors include: a decrease in the estrogen level in women after the menopause, catheterisation, urinary bladder dysfunction, hypertrophy of the prostate gland, diabetes, neurological illnesses. The diagnosis of bacteriuria is based on quantitative urine culture (positive result--> or = 10(5) CFU/ml bacteriae). The most frequent pathogens are: E. coli, enterococci, staphylococci, Pseudomonas aeruginosa, Proteus mirabilis. The antimicrobial therapy is not advised for asymptomatic bacteriuria. In the case of symptomatic bacteriuria it is advised to take urine for culture and to perform sensitivity testing as well as blood culture and to start a "blind therapy". In order to use the antimicrobial treatment effectively, the most frequently occurring pathogens should be registered and their sensitivity patterns in the given hospital recognised. PMID- 12898973 TI - Type 2 diabetes mellitus--a multifactorial disease. AB - Type 2 diabetes is a multifactorial disease caused by both oligo- and polygenic genetic factors as well as non-genetic factors that result from a lack of balance between the energy intake and output and other life style related factors. ADA and WHO have recently reclassified diabetes on both clinical stages and aetiological grounds. There is a lot of data related to the genetics of type 2 diabetes. However, many genes and gene products as well as their interactions with the environment at the molecular, cellular, tissue, and the whole organism levels are still unknown. Changes in the frequency of diabetes occurrence in various urban and rural populations and ethnic groups prove the relationship with the transition to the 'western' life style. Understanding of diabetes pathogenesis is essential to the development of new methods of treatment and strategies of effective prevention of this disease. PMID- 12898974 TI - Primary and specialist diabetes care three years after introduction of health care system reform in Poland. AB - The authors discuss epidemics of diabetes in the world and in Poland. In the Lublin region (eastern Poland), for instance, they found type 2 diabetes (DM 2) in 15.6% of the examined aged over 35 (according to the WHO criteria of 1985). The health care system reform in Poland has made more difficult the access of the diabetic to a specialist that treats this disease. Therefore doctors and nurses of primary health care have become more responsible for diabcare than before. The authors believe that the systematic education of primary health care doctors by specialists so that they can treat patients according to the modern standards of practical diabetology as well as sharing of tasks and responsibilities between primary and specialist diabetologic care, are very important. Primary health care would be in charge of prevention and early diagnosis of DM 2 as well as prevention and early diagnosis of concomitant complications of the disease. Specialists would have consultation on the patients at the moment of diagnosis and then at least once a year. They would also take care of search for and diagnosis of remote diabetes complications. Primary health care doctors would still treat most of diabetics with DM 2; specialist centres doctors would treat most of diabetics with DM type 1, patients with complications and from special risk groups (e.g. women with gestational diabetes). PMID- 12898975 TI - Shaping of psycho-social independence in neurological patients. AB - The aim of the study was to determine the degree of psycho-social independence among neurological patients. The study was conducted by means of the Neurological Patients' Psycho-Social Independence Scale developed by the author, and covered 120 patients--51 males and 69 females aged 21-72; unmarried--34 and married--86, with the level of education: elementary--33, secondary school--79 and university- 8; who had cerebral stroke--67, multiple sclerosis--31, radicular syndrome--22. During the first stage of the study psycho-social independence remained primarily on the medium level with respect to the knowledge concerning life style, apart from the consumption of alcohol, strong tea and coffee, the patients showing a good knowledge of this problem prior to the implementation of the education programme. With respect to skills and the remaining psycho-social aspects (motivation, self-acceptance, social functioning, intellectual efficiency) the respondents represented primarily a very high or high level during the second study. Unfortunately, a considerable number of patients still showed a medium or low level. Population who represented a very high and high levels of psycho social independence, increased by 25.0%. PMID- 12898976 TI - Psycho-social problems of HIV carriers. AB - HIV carrier state and AIDS generate multiple medical, psychological, social and legal problems. The aim of the work was to evaluate selected psycho-social problems among HIV carriers. The examination method was a questionnaire. We interviewed 60 HIV carriers. The results obtained confirmed the existence of the problems affecting the carriers and their families. It creates need for social education in this field. PMID- 12898977 TI - Interaction between local anesthetics and centrally acting antihypertensive drugs. AB - Local anesthetics are widely used in alleviating pain concomitant with small surgery procedures and in dentistry. Especially medication in ductal anesthesia after penetrating into systemic circulation can have significant influence on the central nervous system and stimulus conduction in the heart. Clonidine and reserpine are centrally acting antihypertensive drugs. The aim of this study was to examine an interaction of lidocaine, articaine and mepivacaine with some antihypertensive drugs clonidine and reserpine on the pentylenetetrazole induced seizures. IN CONCLUSION: articaine is the most safe local anesthetic and can be used in epileptic patients Co-administration of local anesthetics with centrally acting antihypertensive drugs did not influence seizures activity in mice. PMID- 12898978 TI - Influence of iodine ions in drinking water on their excretion with urine. AB - The purpose of the work was to appraise the correlation between magnitude of iodine excretion with urine and concentration of iodine ions in drinking water in the population living in the Lublin province. The tests were also performed among the citizens of the Zamosc voivodship. Mean concentration of iodine in urine of the tested citizens was 132.0 micrograms/g creatinine. Mean value of I ions in drinking water was 27.5 micrograms/l. The obtained results of concentration of iodine both in urine and drinking water did not differ as shown in references. Statistically significant correlation between the measure of I excretion in urine and I ions concentration in drinking water was confirmed. There is no ascertained deficiency of iodine in Zamosc region inhabitants. The concentration of iodine in drinking water from adjacent wells was fluctuating in wide limits, but did not differ from recommended norms. There is a positive correlation between concentration of iodine in drinking water and its level in excreted urine. PMID- 12898979 TI - The demand for health education among patients with diabetes. AB - Education of the sick with diabetes has been a stable element of treatment for plenty of years. However, the system of trainings related to this disease is still not perfect and the knowledge of patients about this subject is insufficient. The objective of the study was to determine the demand for health education among patients with diabetes by means of evaluating the level of their knowledge. The study included 130 people suffering from diabetes at the age from 18 to 96 with the disease duration ranging from one year to above 20 years. The level of patients' knowledge about this disease was examined with the use of the survey questionnaire, especially designed for these reasons. The applied survey consisted of 54 questions of which a substantial part was concerned with general information about people suffering from diabetes. Further sections of the survey explored the knowledge about the nature of the disease, its complications, self control, lifestyle and contained the questions concerning the sources of the acquired knowledge on this subject, participation in educational meetings and trainings concerning the issue of coping with diabetes as well as self-evaluation of the level of knowledge. The survey shows that 54.6% of the respondents cannot explain the mechanisms of type 1 diabetes, while 39.2% of the patients do not know the nature of type 2 diabetes. None of the patients was able to enumerate all the symptoms of hypoglycemia and hyperglycemia or the organs and systems most frequently affected by complications. More than 1/3 of the subjects did not know the proper nutrition rules in diabetes. None of the respondents was able to specify correctly all the control examinations and tests that should be taken at least once a year or once every six months. Only 6.1% of the patients evaluated their knowledge about diabetes at a very good degree, while 53.1% of them defined it as good. They were mostly insulin dependent patients under the care of diabetology clinics. However, as many as 40.8% of the respondents considered their level of knowledge unsatisfactory. The research results showed that the leading source of knowledge about the nature of the disease and ways of coping with it were diabetologists (61.5%), followed by nurses (33.8%) and general practitioners (26.1%). The survey results indicate the need for the increased accessibility and intensity of the educational activities in diabetological health care. PMID- 12898980 TI - How soon will we prevent neurodisability in childhood? Symposium proceedings. 14 15 February 2002. PMID- 12898981 TI - Recent trends in neurodisability: implications for research and prevention. PMID- 12898982 TI - Normal and abnormal development of the central nervous system I. PMID- 12898983 TI - Normal and abnormal development of the central nervous system II. AB - In this brief review, I have summarized some of the 'normal' events as found in fetal autopsy material and depicted the prevalence of several specific pathological abnormalities as well as their risk factors. PMID- 12898984 TI - Neurodisability and the channelopathies. PMID- 12898985 TI - Neurodisability: metabolic and hormonal aspects. PMID- 12898986 TI - Studies of children in developing countries. How soon can we prevent neurodisability in childhood? PMID- 12898987 TI - Likely changes in obstetric practice and neurodevelopmental outcome--the next 10 years. PMID- 12898989 TI - Autism: known and unknown. PMID- 12898990 TI - Prevention in severe learning disability: lessons from a follow-up study of young adults with a history of childhood learning disability. PMID- 12898991 TI - Childhood epilepsy: secondary prevention is crucial. PMID- 12898993 TI - Back-seat driver. Interview by Sarah Grant. PMID- 12898994 TI - Knight of data. Interview by Alastair McLellan. PMID- 12898995 TI - Emergency care. Change of pace. AB - A GP practice set up a new system of assessing patients, with an emphasis on appropriate immediate need. The aim is to speed up treatment and reduce inappropriate accident and emergency admissions. A new role of community paramedic on loan from the ambulance trust was a vital part of the team. The first eight months dramatically cut accident and emergency admissions and sped up category A response times. PMID- 12898996 TI - Cancer care. Exchange visit. AB - A trust has set up an ongoing visiting/research cancer programme with two New York Institutions. Notable strengths of US services were greater resourcing and influence of clinicians, better 'pathway' design and increased IT investment. Specific recommendations for the trust include more clinical nurse specialists, more emphasis on research for clinicians and better communication within the network. PMID- 12898997 TI - HSJ people. Directive action. PMID- 12898998 TI - Structural engineering. PMID- 12899000 TI - Head for figures... Assist survey of attitudes to the national programme for NHS IT. PMID- 12899001 TI - Cold front. PMID- 12899002 TI - Ringing the changes. PMID- 12899003 TI - Share alike. PMID- 12899004 TI - [Outlooks for promoting antituberculosis care for the Siberian population]. AB - An analysis of the TB situation in three Siberian territories, i.e. the Kemerovo, Omsk and Tomsk Regions, is presented. Conclusions pertaining to the key specific features were made and a correlation between an epidemic level tension, on the one hand, and an efficiency of the anti-TB activity management, on the other hand, was established. The conclusions were considered as a precondition for setting up a management structure in the sphere of the anti-TB activity that would accumulate the achievements of home phthisiology and of modern methodological and methodical management approaches. Results of elaborations of Novosibirsk Research Institute of TB Issues, which are related with establishing an anti-TB management system in Siberia, are presented; such system would comprise aims, functions, a structure and the key management components. Supposedly, the described approach is expected to optimize the efficiency and good results in the anti-TB management activity at all levels with the resources being distributed and used on the rational basis. PMID- 12899005 TI - [A category of patients with tuberculosis concomitant with HIV infection in an anti-TB facility]. AB - A category of patients with tuberculosis concomitant with HIV infection, who were admitted for inpatient care to the infection department of Tuberculosis Clinical Hospital No. 7, Moscow, during 1996-2001, was analyzed. Peculiarities of the mentioned patients' category (205 subjects) were studied at the anti-TB facility. It was established that males (83.4%), aged 21-30 (48.9%), as well as unemployed (71%) prevailed. As much as 14% of them were homeless and 33% had a prison history. Drug-addiction (76%) and hepatitis C and B (77%) were found to be the key concomitant pathologies in them. HIV was primarily diagnosed at the anti-TB facility in 52% of patients, while tuberculosis had set on before HIV in 34.8% of patients. A major part of patients with tuberculosis concomitant with HIV, who were at the anti-TB facility, had early HIV stages. Specific features of the clinical course of tuberculosis were defined for patients with early HIV stages. It was established that tuberculosis concomitant with early HIV stages is deprived of any peculiarities except for the primary signs' stage, if it has the form of an acute infection. An exacerbation of the tuberculosis process, which quite often leads to its generalization and fatal outcome, can happen during the mentioned period due to a pronounced immunodeficiency. PMID- 12899006 TI - [Experience with using anti-TB preparations (rifabutin)]. AB - The study deals with the first experience of practically using a computer program designed to monitor the treatment process with a subsequent analysis of the efficiency of an anti-TB therapy with regard for different characteristics of patients. The efficiency of chemotherapy was compared (by using the clinical, microbiological and laboratory criteria) in two patients' groups, i.e. a control group (standard chemotherapy regimens) and a group, whose patients received rifabutin. Subgroups were isolated from among the last mentioned patients according to the below factors: primary and secondary therapy courses, completed and interrupted chemotherapy courses, and patients with multidrug resistance who discharged M. tuberculosis. Statistically reliable advantages of rifabutin were shown in respect to the arrest of bacterial discharge during a sufficiently prolonged (at leas 4 months) treatment course applicable to primarily diagnosed patients including those with multidrug resistance and M. tuberculosis. Disadvantages related with the application of rifabutin (a lack of clear-cut indications and abuse of treatment terms virtually in 50% of cases), which reduces the efficiency of its prescription, were equally detected. PMID- 12899007 TI - [Therapy strategy in the treatment of patients with drug-resistant pulmonary tuberculosis]. AB - The article contains a clinical- X-ray- and laboratory description of 56 patients with tuberculosis (disease length ranging from 2 to 5 years), who, during the previous treatment stage, received a therapy in compliance with the generally accepted methods and without any effect. All patients discharged bacteria. Drug resistance to 3 and more preparations was registered in all patients. A complex approach to the therapy of patients with destructive pulmonary tuberculosis is suggested, within the study, with regard for the presence of multidrug-resistance of Mycobacterium tuberculosis (MBT), for a degree of inactivation of isoniazid and for a degree of bacterial-static activity of patient's blood, which provides for a higher treatment efficiency. PMID- 12899008 TI - [Extracorporeal immunopharmacological therapy in the treatment of patients with acutely progressing froms of pulmonary tuberculosis]. AB - Extracorporeal immune-pharmacological therapy, involving Diuciphonium, was added to the complex treatment of 46 patients primarily diagnosed as having acutely progressing destructive pulmonary tuberculosis. Investigations showed that this method sped up the regression of clinical and X-ray signs, during 1 to 2 months, and recovered the immune reactivity in 89% of patients. The clinical changes were dynamically compared with those of 50 other patients in another group (including 39 patients with caseous pneumonia), who were treated by small-volume plasmapheresis. Advantages of applying the extracorporeal immune-pharmacological therapy were demonstrated; they comprise: a shorter time needed to arrest the endotoxicosis, a fuller and faster recovery of the main populations and sub populations of lymphocytes and a less number of fatal outcomes. PMID- 12899009 TI - [Mycobacterium tuberculosis resistance in newly diagnosed patients and in patients with disease relapse]. AB - The frequency rate of resistance of Mycobacterium tuberculosis (MBT) was investigated in 273 patients with primary diagnosis and in 117 patients with relapses, who were registered with tuberculosis of respiratory organs, during 2000-2001, at a TB prophylactic center. Resistance was detected in 12.5 and 17.1% of the obtained cultures, respectively. Mono-resistance was found, from among 54 patients with stable strains, in 25.9% of cases, while multi-resistance, which was detected more often in relapses rather than in persons with primary diagnosis, was found in 50% of cases. The efficiency of treatment depended on a number of preparations, to which there was MBT resistance. The best results were obtained in cases of mono-resistance, the worst ones--in cases of multi resistance. The latter is an indication for changing the standard chemotherapy scheme. The treatment of primarily diagnosed patients turned out to be more effective as compared to that of patients with relapses. PMID- 12899010 TI - [Specific features of tuberculosis of peripheral lymph nodes with specific process in the respiratory organs]. AB - 102 patients with verified tuberculosis of the peripheral lymph nodes (TPLN), including 40 patients with specific process in the respiratory apparatus, were examined. The lymph nodes of the neck were found to be primarily affected in such patients (89%); by the 2nd month, conglomerates develop from such nodes, with the latter evolving into fistulas (44.4%) and into abscesses (40%). A high specificity and sensitivity of the test with the blood serum for adenosine deaminase were proven. Different types of anti-tuberculosis antibodies are isolated with various rates; therefore, it is advisable to study the answers to all serological reactions. PMID- 12899011 TI - [Diagnosis of tuberculosis within the general therapeutic system]. AB - The articles contains an analysis of a dependence of the clinical-pattern nature of primarily diagnosed pulmonary tuberculosis on a tuberculosis form; on a time elapsed from the onset of primary disease signs to the requesting the medical care by a patient; and on a duration of patient's stay in a general therapeutic facility until he or she is transferred to a specialized anti-TB hospital. Medical histories of 177 patients, who were admitted to a regional anti-TB health center, were investigated. The specific pulmonary process was detected, in all patients, upon their asking for medical care within the general therapeutic network. Young patients, aged 20 to 29, were found to ask for medical assistance more often during the first days after the disease onset. Patients after 40 visited their doctors during a month and even more after the appearance of the initial disease symptoms. Pronounced clinical signs of the disease were shown to contribute to an earlier applying for medical care by patients. However, the diagnostics was impaired in pulmonology hospitals due to an intensified nature of clinical syndromes. Such syndromes, when interpreted without paying respect to peculiarities of the disease onset and without a thorough collection of anamnesis data, resulted in the wrong diagnosis. Pneumonia was diagnosed in a majority of patients, i.e. 126 cases (71.2%), in pulmonology department. A long-term stay in a non-specialized hospital and an inadequate therapy, involving large-spectrum antibiotics, aggravated the tuberculosis process. 30 from among 177 patients, who were transferred to the Regional TB health center of the Bryansk city from the general therapeutic facilities, died of the progressing specific pulmonary process. PMID- 12899012 TI - [TNF-alpha in prognosis of a recovery of the spinal marrow function in patients with spinal tuberculosis]. AB - The TNF-alpha significance in forecasting a degree of recovering of the spinal marrow functions was studied in complicated courses of tuberculous spondylitis in 37 patients with generalized and multiple tuberculosis. The TNF-alpha level in the cerebrospinal fluid was found to be related with a severity of inflammation and of neurological disorders, as well as with a degree of spinal marrow compression and with a speed of regression of postoperative disorders in the spinal marrow. The initial TNF-alpha concentration of > or = 400 pg/ml was indicative of a possibility to ensure a fast regression of postoperative disorders in the spinal marrow, while no complete recovery of spinal-marrow functions was observed in cases the TNF-alpha was < 400 pg/ml. PMID- 12899013 TI - [Long-term results of surgical treatment of pulmonary tuberculosis wtih respect to the extent of bacterial isolation and to drug resistance of Mycobacterium tuberculosis]. AB - The remote postoperative results were followed up for as long as up to 10 years. It was established in case of abacillary patients that the frequency rate of relapses was minimal ranging from 3.6% in the group of patients operated for tuberculosis and caseous-necrotic tuberculosis to 11.5% in fibrous-cavernous tuberculosis. Patients with bacterial discharge, primarily of the extensive nature, had more often relapses (19.4% in operated patients for caseous-necrotic tuberculosis and 31.7% in patients operate for fibrous-cavernous tuberculosis). Drug resistance (DR) of Mycobacterium tuberculosis (MBT) to three and more anti TB preparations essentially increase a possibility of relapses in all clinical forms of tuberculosis with maximum threat being in patients with fibrous cavernous lesions (36%). As for the repeatedly operated patients for postoperative relapses involving DR MBT to isoniazid and rifampicin, only slightly above 50% of them survived 5 years after surgery; the index of the 5 year survival was 33% among the patients with DR to 4 and more drugs. PMID- 12899014 TI - [Tuberculosis of respiratory organs concomitant with tuberculosis of bones and joints in young children]. AB - Specific features of thoracic tuberculosis were studied in children with various forms of tuberculosis of bones and joints, i.e. tuberculous spondylitis, arthritis and osteitis. Peculiarities of the clinical course and of complications of thoracic tuberculosis were detected in children with the mentioned clinical forms, which are mostly pronounced in spinal tuberculosis and tuberculosis of joints. A complicated course of thoracic process predetermines the duration of treatment and affects the choice of time for surgical intervention. PMID- 12899015 TI - [Dysfunctions of respiration and blood circulation in patients with concomitant tuberculosis of the spinal column and lungs]. AB - Functional disorders in the respiratory apparatus and blood circulation were studied in 76 patients with tuberculosis of the spinal column concomitant with pulmonary tuberculosis. Spirography, pneumotachography, elctrocardiography, integral rheography and the method variability of cardiac rate were made use of in the diagnostics of the mentioned disorders. Combined tuberculosis of the spinal column and lungs was found to be accompanied, in case of an active pulmonary process, by instable hemodynamic indices, a reduced contractive myocardium function, an insufficient hemodynamic provision and an increased tension in the respiratory system (impaired ventilation and bronchial patency). Simultaneously, a significant vegetative imbalance with an extremely pronounced hypersympathicotonia and over-tension of the adaptation mechanisms in an active pulmonary process, were registered. The detected dysfunctions need to be corrected due to a forthcoming surgical treatment of tuberculous spondylitis. PMID- 12899016 TI - [Osseous-perichondral autoplasty in surgery for tuberculosis of the joints]. AB - The results of 68 reconstruction surgeries made in the hip and knee joints in tuberculosis and concomitant diseases were studied. Advantages of operations involving the component plasty of joint ends (osseous autoplasty, as well as auto perichondral plasty and their variations), including under the conditions of revascularization of the osseous bed, were proven on the basis of the results of a comparative analysis of clinical-and-functional data and X-ray examinations. The use of the method ensuring additional sources for vascularizing the joint tissues improves the conditions of autograft assimilation; the application of perichondral grafting preserves or expands the joint split, which provides an increased mobility amplitude in a damaged joint. The use of perichondral plasty and of its combination with revascularization of the joint osseous joint tissues secured more favorable results as compared to the implementation of osseous plasty interventions alone. PMID- 12899017 TI - [Regimens and efficiency of electromagnetic irradiation in patients with sarcoidosis of the respiratory organs]. AB - The efficiency of different regimens of electromagnetic irradiation therapy (MEIT) was analyzed in cases of 87 patients with primary sarcoidosis of the respiratory apparatus. The study resulted in choosing the optimal MEIT regimens with regard for a form of thoracal sarcoidosis. A high efficiency and safety of MEIT, when applied in patients with sarcoidosis of the respiratory apparatus, as well as its advantages versus systemic corticosteroids were demonstrated. PMID- 12899018 TI - [Drug resistance of Mycobacterium tuberculosis in the North-West of Russia]. AB - The morbidity of primary and secondary drug resistance (DR) of Mycobacterium tuberculosis (MBT) was studied in the North-West of Russia during 1991-2001. The frequency rate and structure of, mainly, secondary DR MBT was investigated in tuberculosis of different localizations on the basis of data provided by clinics of Saint-Petersburg Research Institute for Phthisiopulmonology (PRIP) during 1990 2000. A sharp increase of primary DR MBT (in the North-West of Russia) up to 35 45.5% was detected in the Murmansk, Arkhangelsk and Saint-Petersburg Regions as well as in the Republic of Karelia, an increase of up to 61% was registered in the Kaliningrad Region. The frequency rate of secondary DR MBT went up by 1.5-2 times reaching 80-86% in the Murmansk, Arkhangelsk and Kaliningrad Regions. According to the PRIP clinics, there is also an essential growth of DR MBT in all tuberculosis localizations with a trend of the stability structure shifting towards poly-resistance, which accounts for 65.2% in pulmonary tuberculosis and for 33.6% in extra-pulmonary tuberculosis. PMID- 12899019 TI - [An experimental evaluation of new plastic materials in the surgery of osseous tuberculosis]. AB - Experiments related with xenoplasty made in 108 rabbits are described. An original method of processing the spongy xenografts is suggested; it provides for both minimizing the immune response and for obtaining a new plastic material applicable for replacement of the postoperative osseous cavities. A complete healing of the operated bone and a close-to-norm recovery of osseous structures were ensured in experiments with animals. A similar result was obtained in experimental osseous tuberculosis after the removal of tuberculous foci. The study materials are passed over for testing in clinical practice. PMID- 12899020 TI - [A case of multiple operations in pulmonary tuberculosis]. PMID- 12899021 TI - [Prevention of adverse reactions of antituberculous drugs in pulmonary tuberculosis in patients with different genetic background]. AB - A hundred and sixty-seven patients with pulmonary tuberculosis were examined to study the genetic predisposition to adverse reactions (AR) from chemical drugs. The phenotype of haptoglobin carriage, the type of, the activity of the enzyme glucose-6-phosphate dehydrogenase inactivation were used as a genetic marker. If with a poor combination of the above parameters AR developed in 100%; with their relative combination, it occurred in 78%. With other types of their combinations, AR took place only in 38%. A new method for preventing AR in pulmonary tuberculosis is proposed by using non-traditional methods, such as phytotherapy alone and in combination with ultraviolet autoblood irradiation. The use of this method is shown to reduce the incidence of AR, to attenuate their severity and clinical manifestations particularly in patients with a relatively poor genetic background. PMID- 12899022 TI - [Tuberculosis in Siberia in early 21st century]. PMID- 12899023 TI - Special action projects for the elimination of leprosy. PMID- 12899024 TI - Chronology of travel recommendations, areas with local transmission. PMID- 12899025 TI - [Who is to decide in health care? A psychological deadlock between physicians and administrators]. PMID- 12899026 TI - [Increased risk of cardiovascular diseases and diabetes in mentally ill persons. Life style is a contributing factor]. PMID- 12899027 TI - [Audit of prescribing practices in nursing homes. Many preparations but few serious interactions]. AB - Information on all prescribed drugs to 112 patients in three nursing homes in the community of Malmoe and Trelleborg were analysed. The age of the patients was 84.5 years (mean) and number of prescribed drugs was 9.6 (mean). A computer program was used to analyse any interactions according to the Swedish Drug Compendium (FASS). Among 74 potential interactions only four were classified as clinically important, some of doubtful value in view of new data. Use of psychotropic drugs were investigated and judged for their appropriateness. Although 28% were given neuroleptic drugs, prescribed doses were moderate. To improve treatment and monitoring of antidepressive drugs, the need for use of rating scale for depression became obvious and was planned. To be worthwhile, any intention to change chronic drug therapy of patients in nursing homes must be carefully monitored for clinical effects over a sufficient time. PMID- 12899028 TI - [A chest tube can be inserted safely utilizing standard technique. Minithoracotomy should not be performed routinely]. AB - The National Swedish Board of Health and Welfare has concluded that a minithoracotomy should be used for the insertion of a chest tube. Our opinion is that a less than 2 cm skin incision followed by blunt dissection through the chest wall is equally safe provided that the anatomical landmarks of the safe triangle are identified and the lung is not adherent to the chest wall. A prerequisite for safe treatment is an examination of the chest x-ray, supervision of chest tube function and the patient. Chest tube malfunction or complications should be managed urgently. PMID- 12899029 TI - [An unusual case of a side effect. Cefotaxime-induced confusion in a patient with renal failure]. AB - Third-generation cephalosporins in general have few adverse effects and cefotaxime (Claforan) particularly is considered to be a good choice because of the favourable side effect profile. In patients with severe renal failure there have been reports of confusion and psychosis. The manufacturer therefore recommends that half the ordinary dose should be given to patients with severe renal failure. Half the ordinary dose can still be too much. We describe a patient in hemodialysis who reacted with a reversible encephalopathy with psychosis in spite of reduced doses of cefotaxime. The plasma concentration of cefotaxime was high and the reaction was diagnosed as a dose dependent side effect. PMID- 12899030 TI - [Physicians do not longer rule the health care-roost. Good cooperation between the team and the teamleader is more and more important]. PMID- 12899031 TI - [Prohibited marketing and shift in meaning of indications--a case for the IGM]. PMID- 12899032 TI - [Many physicians gave their lives to solve the mystery of yellow fever]. PMID- 12899033 TI - [About the protective effects of alcohol against cardiovascular diseases: no reason to increase drinking habits]. PMID- 12899034 TI - [A final reply on antihypertensive agents: we want a more powerful authority!]. PMID- 12899035 TI - [Snuff is less dangerous than smoking for the cardiovascular system. Wrong to hide the scientific basis which does not confirm the hypothesis]. PMID- 12899036 TI - [Evidence-based psychiatry, sure--just as we do not stop to think!]. PMID- 12899037 TI - [Psychiatry does need the professional conversation]. PMID- 12899038 TI - [A physiotherapist/scientologist encouraged young people to cease medication with antidepressive agents]. PMID- 12899039 TI - [What information do patients receive prior to randomized surgical studies?]. PMID- 12899040 TI - [Do you treat patients with Seroxat?]. PMID- 12899041 TI - [The National Board of Health and Welfare: in agreement on the chest tube insertion technique]. PMID- 12899042 TI - ["Children's hospital" in TV channel 4, public education and ethical rules]. PMID- 12899043 TI - [Aikido in the dentist's chair]. PMID- 12899044 TI - [Anxiety in the dental office: theoretical aspects and treatment strategies]. AB - Although mild tension and fear when visiting the dentist are considered normal, some patients experience extreme anxiety for dental treatment, which interferes with treatment and oral health, and causes significant distress. The present article reviews the literature on the definition and occurrence of dental fear. Causal and maintenance factors of dental fear are identified and a conceptual model of these factors is presented. From this model several interventions are presented that can be used to treat moderately anxious patients and pathological dental fear. The effects of these treatments are discussed. PMID- 12899045 TI - [Is anxiety your master or are you the master of your anxiety?]. AB - By means of four cases the author demonstrates how he manages with varying success dental anxiety. There is a call for dentists willing to invest time and teach patients how to deal successfully with such anxiety. This activity is part of the dentist's competence, with back support of a supervising psychologist or therapist. PMID- 12899046 TI - [The treatment of anxiety in handicapped persons]. AB - Having worked as a dentist in an institution for handicapped people for 25 years, the author wants to share some of his experiences. By means of the three cases presented here, the author tries to explain that although each mentally disabled patient has a different character, an equal strategy of treatment can be used. PMID- 12899047 TI - [Hypnosis technics used to diminish anxiety and fear: review of the literature]. AB - Hypnosis can be a valuable technique in the management of patients who fear medical treatment. Hypnosis leads to a stronger concentration and a more focussed attention, and thus to a better acceptance and greater effectiveness of suggestions. Literature indicates that hypnotherapy can reduce pain and fear in case of dental or medical phobia. After a short review of the existing literature on the topic, the practical aspects of the hypnotic approach are explained. PMID- 12899048 TI - [Hypnosis and hypnotic phenomenon. Preliminary aid in cases of stress for the patient and dentist]. AB - Dental treatment is often as stressful an experience for the dentist as it is for the patient. Some examples of hypnotic communication and hypnotic phenomena are given, and their value in dental practice discussed. With hypnosis, treatment is possible in a creative and relaxing way, and this reduces stress for dentist and patient alike. PMID- 12899049 TI - [Hypnosis and dentistry: water and fire?]. AB - At first sight a combination of hypnosis and dentistry may very well appear not to be evident at all. Nevertheless, it works quite well in the everyday dental practice. In this article the author provides an overview of indications in which hypnosis can be used. Anxiety and pain are among the most common indications. Hypnotic techniques in dental practice can be explained at two levels: as a general way of communication between the dentist and his or her patient. Or as an advanced technique in the treatment of severe anxiety. The author discusses the several stages related to the latter technique: the intake interview, the induction, the deepening, the therapeutic stage and the deduction. The author also briefly mentions the disadvantages of hypnosis, but on the other hand encourages every dentist to become a hypnodontist. Not only for the benefit of his patient but also, and not the least, for his or her own well-being. All the more because hypnosis has become a scientific issue involving serious research at many renowned universities, including in Belgium. PMID- 12899051 TI - [Too few or too many nursing personnel?]. PMID- 12899050 TI - [Eyes here and there]. PMID- 12899052 TI - [Work and health screening]. PMID- 12899053 TI - [Learning requirements and professional life]. PMID- 12899054 TI - [Application of qualifications for clients]. PMID- 12899055 TI - [Where does the path lead?]. PMID- 12899056 TI - [How ideas also define claims]. PMID- 12899057 TI - [Nurses and the conflict between ethics and economy]. PMID- 12899058 TI - [Help in mediation, constructive conflict and solutions]. PMID- 12899059 TI - Insuring satisfaction. Health plans are steadily upgrading their customer relationship management capabilities. PMID- 12899060 TI - In touch with your assets. Software helps manage equipment, meet regulations and organize maintenance staff. PMID- 12899061 TI - Drug safety for the long-term. Long-term care facilities have the same medication management concerns as hospitals, plus others. PMID- 12899062 TI - The electronic option. A clinical messaging system smooths many paths. PMID- 12899063 TI - Creating a special user experience. Web-based benefits communication offers real value--if members fully utilize it. PMID- 12899064 TI - [A short summary. The history of glucocorticoids]. PMID- 12899065 TI - [On genomic and nongenomic effects. Molecular basis of glucocorticoid action]. PMID- 12899066 TI - [What happens in the body with glucocorticoids? Pharmacokinetics and pharmacodynamics]. PMID- 12899067 TI - [Systemic therapy with glucocorticoids. Treatment of rheumatic diseases]. PMID- 12899068 TI - [Bronchial asthma and inflammatory dermatoses. Topical glucocorticoid treatment]. PMID- 12899069 TI - [Treatment of bronchial asthma. Dosage forms for inhalational glucocorticoids]. PMID- 12899070 TI - [Between Cortisone anxiety and euphoria. Pharmaceutical surveillance of long-term medication]. PMID- 12899072 TI - [Integrated functions of the nervous and endocrine systems]. PMID- 12899071 TI - [Solid organ transplantation. Glucocorticoids in kidney transplantation]. PMID- 12899073 TI - Tubulointerstitial dysfunction in early post-transplant period. AB - Dysfunction of a transplanted kidney may develop at any time in the post transplant period. The aim of this study was to differentiate levels of early dysfunction of a transplanted kidney. The study included 45 examinees undergoing kidney transplantation. They were divided into four groups, in regard to length of hospitalization and post-transplant complications: group I (up to 15 days, complication-free); group II (up to 15 days, with complications); group III (up to 30 days); group IV (up to 60 days). The control group included patients undergoing abdominal surgery, without uropoetic system disorders. The following parameters were examined on a daily basis a month after transplantation on average: creatinine clearance, creatinine and urea. Statistical analysis of these parameters revealed the following levels of renal dysfunction: control group- circulatory tubular dysfunction without azotemia; group I--polyuric acute tubular necrosis; group II and group III--severe or moderately severe polyuric acute tubular necrosis and group IV--polyuric acute tubular necrosis. PMID- 12899074 TI - [A successful population-based method for smoking cessation]. AB - INTRODUCTION: Our country is in the third place in Europe concerning tobacco smoking. Although strict law regulations regarding indoor smoking have been brought, the law banning all tobacco advertising, and the behaviour of our population are inadequate. Our objective was to persuade smokers for the "Quit and Win" campaign and to establish the number of smokers in health personnel employed in health facilities. MATERIAL AND METHODS: This population-based smoking cessation campaign was coordinated by Federal Institute of Public Health, through a network of Public Health Institutes within the country with the support of national and local media. RESULTS AND DISCUSSION: Quit and Win campaign was organized for the third time. The campaign was realised with the financial support and sponsorship at community level throughout the country. The Federal Ministry provided a national health award. The national and local media accompanied the campaign. The campaign included 3.178 smokers and 2.575 supporters, that is 0.1% of the population over 18 years of age. This is in accordance with participants in some other countries, who had a better support. More than 60% of health care facility employees are smokers. CONCLUSIONS: Our tradition, habits in the society and overall situation encourage smoking habits to spread in general population. Our campaign has proved that people should be motivated to quit smoking, but they need to be informed. Actions taken in general population and based on a positive smoking cessation program in which smokers are willing to stop smoking have given unexpectedly good results. PMID- 12899075 TI - Contralateral acoustic suppression of transient evoked otoacoustic emissions- activation of the medial olivocochlear system. AB - Medial olivocochlear pathway represents the final part of efferent acoustic pathway which comes from the superior olivary complex ending at outer hair cells. Activation of medial olivocochlear system (MOCS) alters the cochlear output decreasing the travelling wave within cochlea. Stimulation of MOCS provides protection against moderate levels of noise, encoding noise signals as well as selecting hearing attention. Activation of MOCS can be performed using contralateral acoustic stimulation. The principal result of presentation of contralateral acoustic stimulation during screening of transient evoked otoacoustic emission (TEOAE) is an attenuation of the TEOAE amplitude. Thirty eight ears were examined in this study: twenty-eight ears from 14 normal-hearing adults and 10 patients with unilateral deafness. Healthy subjects were exposed to contralateral broad-band noise of various intensities (40, 30, 20 and 10 dB SL), as well as 30 dB SL pure tone stimulation (1 kHz and 4 kHz). A decrease of TEOAE amplitudes during contralateral stimulation with 40 and 30 dB SL broad-band noise and pure tones was established. This effect was a result of MOCS activation. A greater intensity of contralateral stimulation evoked greater decrease of TEOAE amplitude; stimulation with broad-band noise caused greater attenuation than with pure tone stimulation. Contralateral stimulation of deaf ears in the group with unilateral deafness was also performed. Statistically significant difference between TEOAE amplitude before and during contralateral stimulation was not established. This circumstance explains that activation of MOCS and consequent reduction of outer hair cells motility is very possibly caused by contralateral acoustic stimulation. Apart from studying physiological significance of efferent auditory system, results of this and similar studies can be used for production of hearing aids improving speech discrimination in noisy environment. PMID- 12899076 TI - [Serum copper levels in early pregnancy complicated by symptoms of spontaneous abortion and infection]. AB - INTRODUCTION: The study evaluates serum levels of copper, chorionic gonadotropin, estradiol, progesterone and prolactin in patients with symptoms of miscarriage and in uncomplicated pregnancies in cases with or without cervical or vaginal infections detected by vaginal or cervical smears, as well as Chlamydia testing. MATERIAL AND METHODS: The study included 50 patients with symptoms of threatening miscarriage and 50 patients with uncomplicated pregnancies. Hormone levels were determined by ELISA method and copper was evaluated by acid medium colorimetry. RESULTS: We found that values of serum copper, estradiol, progesterone and prolactin were significantly lower in patients with lower genital tract infection. CONCLUSION: Decreased levels of serum copper could be used as a method of choice for detecting infection during the first trimester of pregnancy. PMID- 12899077 TI - [Low level laser irradiation and its effect on repair processes in the skin]. AB - INTRODUCTION: Application of laser beams for therapeutic purposes is of relatively recent date, but today there is no field of medicine where lasers cannot be used. PHYSICAL CHARACTERISTICS OF LASER RADIATION: Laser radiation is a type of electromagnetic radiation with some specific characteristics such as coherence, monochromaticity and parallelity. TYPES OF LASER DEVICES: Nowadays, there are many laser devices on the market used in medicine and dentistry. According to the type of their active medium, lasers can be classified as solid, gas, semiconductor and liquid. EFFECTS OF LOW LEVEL LASER THERAPY ON BIOLOGICAL SYSTEMS: The exact mechanism of action of low level laser therapy is still not completely understood. Its basic feature is to modulate cell behaviour, without causing significant temperature increase. During irradiation of a tissue with a laser beam, an interaction between cells and photons takes place--photochemical reaction. After a cell absorbs the photon, the photon stops existing, and its energy is incorporated into the molecule which has absorbed it. Once this energy is transferred to different biomolecules, it can be transferred to other molecules as well. The energy transferred to the molecule can increase its kinetic energy, and activate or deactivate enzymes or alter physical or chemical properties of main macromolecules. EFFECTS OF LOW LEVEL LASER THERAPY ON WOUND HEALING: Effects of low level laser therapy on wound healing process is one of the most fully studied aspects of this type of therapy. It affects all phases of this very complex process. This paper offers a more detailed analysis of these aspects. PMID- 12899078 TI - [Development and differentiation of adipose tissue]. AB - INTRODUCTION: For years adipose tissue has been considered inert, serving only as a depot of energy surplus. However, there have been recent changes, undoubtedly due to advancement of methods for studying the morphology and metabolic activities of adipose tissue (microdialysis and adipose tissue catheterization). In normal-weight subjects, adipose tissue makes 10-12% with males and 15-20% with females. About 80% of adipose tissue is located under the skin, and the rest envelops the internal organs. With humans there are white and brown adipose tissues, which is predominant with infants and small children. HISTOLOGIC CHARACTERISTICS: From a histological point of view, it is a special form of reticular connective tissue, which contains adipocytes with netlike structure. Human adipose tissue has four types of adrenergic receptors with different topographic dispositions, which manifest different metabolic activity of adipocytes of particular body organs. Changes in adipose tissue are associated with the process of adipocyte differentiation. Critical moments for this process are last months of pregnancy, the first six months of infancy and then puberty. However, the differentiation process may also begin during maturity. Namely, as size of adipocytes can increase to a certain limit, this process can be activated after reaching a "critical" adipocyte volume. The differentiation process is affected by a number of hormones (insulin, glucagon, corticosteroids, somatotropin (STH), thyroid gland hormones, prolactin, testosterone), but also by some other substances (fatty acids, prostaglandins, liposoluble vitamins, butyrate, aspirin, indomethacin, metylxanthine, etc.). PMID- 12899079 TI - [Disorders in fracture healing]. AB - INTRODUCTION: Bone fracture healing is a complex cascade of events at cellular and biochemical levels, that ends by complete structural and functional restoration of a damaged bone. Impaired healing develops in 5-10% of all fractures, and manifests as delayed union or non-union. This paper deals with the problem of impaired healing as well as with methods of fracture healing enhancement. CAUSES OF IMPAIRED FRACTURE HEALING: There are many factors causing impaired fracture healing (inadequate vascularization, mechanical causes, infection, etc.), and it is very important to recognize the principle cause of delayed union and non-union, since therapy is based on eliminating the factor that causes it. FRACTURE HEALING ENHANCEMENT: Through constant attempts to find adequate solutions and procedures in order to resolve the problem of impaired fracture healing, many alternatives in treatment of impaired healing have been developed. Some of these procedures may also be useful in treatment of fresh fractures, especially when it comes to fractures that are prone to delayed union and non-union more than usual. All currently known methods of healing enhancement may be classified as biological, mechanical and biophysical. CONCLUSION: Certain methods are in clinical use for several decades. The newest methods, such as locally applied growth factors, composite biosynthetic grafts, gene therapy and systemic approaches are studied all around the world, and are on the verge of clinical application. Due to impressive number of therapeutic options, certain therapeutic procedures of choice will be developed for specific impairments. PMID- 12899080 TI - [High thoracic epidural anesthesia in coronary surgery]. AB - INTRODUCTION: High thoracic epidural anesthesia and analgesia are being increasingly used for coronary artery bypass graft surgery. The reasons for this include excellent perioperative pain control with advantage of early tracheal extubation, improved postoperative pulmonary function, and cardiac protection due to sympthatetic blockade. EFFECTS OF HIGH THORACIC EPIDURAL ANESTHESIA: Cardiac protection is the consequence of decreased heart rate, myocardial contractility and arterial blood pressure, without changes in coronary perfusion pressure. Therefore, high thoracic epidural analgesia beneficially alters major determinants of myocardial oxygen consumption, without jeopardizing coronary perfusion pressure. In addition, decrease of functional residual capacity, may reduce postoperative pulmonary morbidity. RESULTS OF CLINICAL STUDIES: Patients with high thoracic epidural anesthesia revealed a more favourable perioperative hemodynamic profile, lower incidence of ischemia and better response to perioperative stress. HIGH THORACIC EPIDURAL ANESTHESIA TECHNIQUE: The epidural catheter should be placed at the Th2/Th3 interspace at least one hour before administration of heparin. After local anesthetic bolus dose, a continuous epidural infusion is recommended. CONCLUSION: There is strong evidence for beneficial effects of high thoracic epidural anesthesia in patients undergoing surgical myocardial revascularization. However, it is still underutilized in current clinical practice. PMID- 12899081 TI - [Ovarian epithelial carcinoma--a malignant disease sparing no age group]. AB - INTRODUCTION: Ovarian epithelial carcinoma is one of the most common gynecologic malignancies and the fifth most frequent cause of cancer death in women. Ovarian cancer affects women 65 years of age and older more frequently than younger women. MATERIAL AND METHODS: Four young patients with ovarian epithelial carcinoma were treated at the Institute of Oncology in Sremska Kamenica, Serbia and Montenegro, in the period 1981-2000. The median age at diagnosis was 17.3 years. Final histopathological findings revealed cystadenocarcinoma, either mucinous or serous. The stage of the disease was established using International Federation of Gynecology-Obstetrics (FIGO) Calssification. Spread of the disease dictated the extent of operation and post-operative chemotherapy. Patients were strictly followed-up at the Institute. RESULTS: The first two patients underwent unilateral adnexectomy as initial treatment. Final histopathological examination revealed an epithelial ovarian carcinoma, stage IIIa and IIb. Total hysterectomy with unilateral adnexectomy and total omentectomy were performed in both patients as second treatment with chemotherapy, according to the Cisplatin/Carboplatin and Cyclophos-phamide (CP) protocol following surgery. Another two patients underwent total hysterectomy with bilateral adnexectomy and total omentectomy as initial treatment with chemotherapy, CP protocol, following surgery. Both patients had stage IIc. Despite treatment, in two patients with stage IIIa and IIc, metastases were diagnosed. In 2 patients survival was over five years. One patient lived less than 3 years, and one less than 4 years. DISCUSSION: Ovarian carcinomas are difficult to diagnose at early stage. Histologic confirmation of the diagnosis, surgical staging, and aggressive surgical debulking, when possible, are all part of the initial evaluation and treatment. In most cases, surgery is followed by chemotherapy. The risk increases from 15.7 to 54 per 100,000 in the age group 40 79 years. Our study included 4 patients, medium age 17.3, with epithelial ovarian carcinoma which warns us to think twice when we get an adolescent patient with an adnexal mass. CONCLUSION: Advancing age, the major risk factor for development of ovarian carcinoma is, of course, unalterable. We investigated 4 patients medium age 17.3 years, with epithelial ovarian carcinoma. Pelvic masses found in women of reproductive age, must also be evaluated preoperatively to determine the probability of malignancy. PMID- 12899082 TI - [Tonsillectomies and adenoidectomies over a 60-year period (1939-1998) at the Otorhinolaryngology Department in Subotica]. AB - INTRODUCTION: This paper presents available epidemiological data on number and types of surgeries of Waldeyer's ring over a sixty-year period, as well as their interactions. RESULTS: Out of 80.477 patients operated on in the period from December 1, 1939 to Dec 31, 1998, statistical survey included 56.564 patients (Table 1). 26.808 (46.57%) patients were operated in local anesthesia, 23.833 (41.40%) under ether narcosis, and 6.923 (12.026%) patients were operated on under general endotracheal anesthesia. 38.846 (67.48%) tonsillectomies, 12.864 (22.34%) tonsilloadenoidectomies and 5.854 (10.16%) adenoidectomies were performed. DISCUSSION: The percentage of these surgeries is very high, but in the past ten years it has fallen below 50%. General endotracheal anesthesia is being used at the Department since 1966, and ether anesthesia is not being used since 1991. We believe that the number of tonsillectomies is high, because it also includes surgeries performed under local anesthesia, in patients over ten years of age. However, we can conclude that anesthesiological procedures effectively followed trends of medical science. PMID- 12899083 TI - [Use of self-reported values of body weight and body height in nutritional status evaluation in adolescents]. AB - INTRODUCTION: The aim of our study was to assess validity of self-reported body weight and height compared to measured values in a group of adolescents. We also investigated differences in reported and measured values and factors responsible for them in this particular population. MATERIAL AND METHODS: 295 adolescents aged 11-14 (114 boys and 131 girls) attending primary schools in central Belgrade, recorded their BMI and completed a series of questionnaires measuring body satisfaction and eating patterns. All participants were measured afterwards. RESULTS: Differences between reported and measured weights were highly significant in girls (48.84 +/- 8.42 kg vs 49.69 +/- 9.28 kg). Significant differences between reported and measured weights were established in the group of adolescents with weight problems (-3.18 +/- 4.03 kg). Adolescents on diets presented a difference of -1.36 +/- 3.36 kg. Weight problems (OR 3.72, 95%CI: 2.10-4.75) and female gender (errors occurred more often for 22%, 95%CI: 0.23 1.80) were significant predictors in multivariate regression model for discrepancy of reported and measured weights. In a multivariate regression model, where differences in BMI were dependent variable, weight category was a significant predictor, with differences being 1.65 times more common (95%CI: 1.13 2.11). DISCUSSION: Our results suggest that self-reported body weight and height have limited reliability and can be used only in normal weight male adolescents. This method should not be recommended in surveys that involve overweight and/or obese adolescents and those who are on diet. PMID- 12899084 TI - [Distribution of the ABO, Rh, MNSs, Kell and Duffy blood group antigens in the population of Vojvodina]. AB - INTRODUCTION: Analysis of erythrocyte blood group antigen polymorphisms and genetic variability in population of Vojvodina was performed by investigating gene and genotype frequencies which determine antigens of ABO, Rh, MNSs, Kell and Duffy blood-group systems. MATERIAL AND METHODS: We investigated 350 unrelated persons from Vojvodina in regard to appurtenance of ABO, Rh, MNSs, Kell and Duffy blood-group systems. We calculated gene, genotype, phenotype frequencies and proportion significance test. RESULTS AND DISCUSSION: Results of investigation revealed that gene and genotype frequencies of investigated blood-group systems are similar to corresponding data for majority of European populations, while statistically significant differences were established in inhabitants of geographically distant regions. Values of proportion significance test revealed statistically significant differences of genotype frequencies for ABO and MNSs blood-group antigens in populations of: Australian Aborigines, Chinese population, Arabians, Blacks, Eskimos, American Indians (Navaho and Pueblo) and population of Papua New Guinea. Statistically significant differences of genotype frequencies were established in inhabitants of narrow geographical areas of Europe such as: Finland, Germany, Sweden, Albania, England and Netherlands. CONCLUSION: Our results point to the fact that erythrocyte blood-groups have different frequencies in some parts of the world, and that there are great differences in frequencies of some blood-groups among inhabitants of various continents and races. Genetical peculiarity of the population of Vojvodina points to the fact that differences in blood-group frequencies are also present among inhabitants of narrow geographical areas. PMID- 12899085 TI - [Role of intravenous cholangio-cholecystography in the diagnosis of asymptomatic choledocholithiasis]. AB - INTRODUCTION: Standard diagnostic procedures (anamnesis, physical examination, laboratory analyses, ultrasound diagnosis), commonly used in diagnosis and preparation for surgical intervention of patients with cholelithiasis, are in most cases a reliable indicator for evaluation of the disease and conditions planned for surgery. DISCUSSION: In some cases by application of these narrow diagnostic models, some conditions, anatomic variations and biliary tract malformations remain unrecognized. Asymptomatic ("silent") choledocholithiasis (2.02%) represents a special diagnostic and therapeutic problem. CONCLUSION: Our extended diagnostic protocol includes routine intravenous cholangio cholecystography as a standard diagnostic procedure for evaluation of cholecysto choledocholithiasis prior planning cholecystectomy. PMID- 12899086 TI - [Persistent hyperemesis gravidarum as a psychosomatic dysfunction--case report]. AB - INTRODUCTION: Persistent severe vomiting in pregnancy is called hyperemesis gravidarum. The etiology of this disorder is still unknown. However, endocrinological disorders and psychological factors are mentioned as possible causes. Vomiting usually stops after the end of the first trimester of pregnancy. CASE REPORT: A case of persistent, severe vomiting during pregnancy is reported. The patient was admitted to the Department of Gynecology and Obstetrics in Novi Sad in the 29th week gestation due to persistent vomiting from the beginning of pregnancy. A psychological evaluation including current psychological status was performed. The obtained results showed that hyperemesis gravidarum was a consequence of associated psychosocial factors, and conflicts causing somatic disorders. CONCLUSION: Such cases point to the importance of psychotherapy in pregnancy either with or without drug therapy. In addition, we underline the importance of psychological preparation before pregnancy and better information about pregnancy and motherhood. PMID- 12899087 TI - [Telemedicine and cardiology]. AB - INTRODUCTION: Telecommunications and information technology provide clinical care at distance by means of telemedicine. Hospitals and other health care facilities use medical telemetry devices for monitoring patients' vital signs. These portable devices are used for measuring patient vital signs such as ECG, blood pressure, heart rate, respiration, capnography (CO2) and other important parameters and then transmit these information to a remote location using a nearby receiver. APPLICATION OF TELEMETRY: Eliminating the need for wired connection with the patient, monitors allow, otherwise bedridden patients to be mobile, which shortens the recovery time. Wireless technology is also useful in the emergency care units, because emergency physicians need not leave their patients while consulting a handheld wireless device. This equipment also enables a paramedic to communicate with emergency physicians for early assessment, well before patients' arrive in hospital. TELEMEDICINE: Certain types of medical telemetry devices may be used in home conditions. Telemetry can provide monitoring and home health care services at distance, using advanced telecommunications and information technology in patients with reasonably stable, but a severe, chronic, difficult condition and caring home environment. This information can enable health-care providers to effectively manage treatment without need for acute emergency treatment and hospitalization. CONCLUSION: Hospitals worldwide are under constant pressure to decrease healthcare cost and to improve treatment outcome. Telemedicine and home health care may be one of the solutions for the problem. PMID- 12899088 TI - [Development of endocrinology in Serbia]. AB - The beginnings of endocrinology in Serbia date back to the second half of the 19th century. At that time some empiric case reports were published, mostly in Serbian Archives of Medicine and today we know that they belonged to the field of endocrinology. Between two World Wars professional articles and more serious studies in endocrinology were also published, but sporadically and non systematically. Rapid progress in endocrinology in Serbia occurred after the Second World War, in many segments reaching European and World levels. Enormous contribution to development of clinical endocrinology was made by Prof. Dr. Mioljub Kicic, Prof. Dr. Dusan Duric and Academician Prof. Dr. Borislav Bozovic, and to basic, experimental endocrinology by Academician Prof. Dr. Radivoj Milin. Apart from chronological reviews concerning foundation of certain endocrinology clinics and institutes in Belgrade, this paper also deals with foundation of endocrinology institutions in Novi Sad and other university centers in Serbia. PMID- 12899090 TI - Geisinger shifts from global to primary care capitation. PMID- 12899091 TI - Capitated groups track claims, reduce costs using MD Hawkeye. AB - The "MD Hawkeye" software program from Doctor-Driven Systems helps capitated groups manage costs, reduce utilization, and improve quality measurements. See how one capitated group saved more than $100,000 on one project. PMID- 12899092 TI - Capitated groups increasingly incentivized to boost vaccinations. PMID- 12899093 TI - New CA law protects capitated providers against rising specialty pharmacy costs. PMID- 12899094 TI - [Screening for Down syndrome using triple marker testing in the second trimester of pregnancy]. AB - The aim of this study was to check the validity of the biochemical screening of pregnancies with Down's syndrome during the second trimester of pregnancy, in order to reduce the incidence of invasive diagnostic procedures. We used the optimal balance between sensitivity and specificity to determine the "cut off" values to estimate the results of the biochemical screening. Between January 1995 and December 2000, 2000 pregnancies were checked by double (determining hCG and AFP serum levels) and triple test, (determining hCG, AFP and uE3 serum levels). Competitive radioimmunochemical procedures (2nd trimester Amerlax-M, Ortho Clinical Diagnostics, USA) were used. The risk of Down's syndrome was calculated by Prenata program (Ortho Clinical Diagnostics, USA). The "cut off" median MoM values in pregnancies with Down's syndrome were 0.73 (AFP); 2.02 (hCG) and 0.74 (nE3). The calculated risk was compared with possibility 1:300 to estimate the results of biochemical screening. Our results were checked in the cytogenetic laboratory where samples of amniotic fluid, that we also took, were sent. We observed lower AFP levels (0.96 +/- 0.09 MoM), uE3 levels (0.65 +/- 0.1 MoM) and higher levels of hCG (1.57 +/- 0.27 MoM) in pregnancies with Down's syndrome, in comparison with euploid pregnancies of the corresponding gestational age. With 1:200 risk, the sensitivity of triple test is 80%, with acceptable number of false-positive results. This cut-off value showed to be acceptable for separating positive from negative results. Invasive procedures should be performed in pregnancies with positive screening result, with the aim of getting the tissue sample of the fetus for further cytogenetic analysis. PMID- 12899095 TI - [The Mediterranean diet and occurrence of malignant tumors of the digestive system in the Croatian islands]. AB - The aim of this study was to compare the age-standardised incidence rates of gastric bowel and pancreatic cancer between two regions of the Republic of Croatia: (1) five middle Dalmatian islands (Brac, Hvar, Korcula, Vis and Lastovo) with "Mediterranean" nutrition; (2) the control population of Koprivnicko krizevacka County (KKC) with "continental" nutrition. The characterisation of nutrition habits in the two study regions revealed very significant differences. The newly registered cases of gastric, bowel and pancreatic cancer between 1986 and 1995 were retrieved from the Cancer Registry of Croatia. The resulting incidence rates were standardised to the standard world population using the method of direct standardisation. Age-standardised incidence rates in the islands in comparison to the control KKC population per 100,000 inhabitants were 17.2 vs. 39.4 (p < 0.001) in males with gastric cancer, 9.1 vs. 16.5 (p < 0.01) in females with gastric cancer, 34.5 vs. 31.4 (p > 0.05) in males with bowel cancer, 18.3 vs. 20.3 (p > 0.05) in females with bowel cancer, 5.5 vs. 9.0 (p < 0.05) in males with pancreatic cancer and 2.7 vs. 5.0 in females with pancreatic cancer (p < 0.05). Standardised incidence rates of gastric and pancreatic cancer in the island populations were significantly lower from the standardized Croatian average. The study showed that "Mediterranean" type of diet could have a protective effect against gastric and pancreatic cancer. PMID- 12899096 TI - [Occult papillary carcinoma of the thyroid gland with unusual neurologic symptoms]. AB - We present a patient who had progressive cognitive decline and Jackson epileptic seizures and CT scan of nonspecific multiple cortical lesions. Post mortem examination revealed multiple cortical metastases of the brain and primary occult papillary carcinoma of the thyroid gland. Solitary metastases of the left kidney and pulmonary lymphangitic spread were also present. Multiple cortical metastases of the brain from primary occult papillary carcinoma of the thyroid gland with such neurological symptoms have not been reported previously. PMID- 12899097 TI - [Evaluation of genetic causes of mental retardation]. AB - Causes of mental retardation that affects 1-2% of general population are numerous and heterogeneous and include genetic and environmental factors. Improvement of diagnostic techniques and progress made in mapping genes associated with specific mental retardation syndromes provide the possibility to make precise diagnosis in a proportion of mentally retarded individuals. The present study reviews the current diagnostic possibilities, highlights the problems involved in the diagnosis of mental retardation, providing practical guidelines for rational diagnostic approach and work up in individuals with mental retardation. Diagnosis is highly dependent on a detailed and comprehensive family and personal history, careful physical examination and long-term follow up of the children with developmental delay. Referral to specialised genetic units, where extensive evaluation based on rational selection of the specific investigations is possible, yields best diagnostic results. PMID- 12899098 TI - [Renal dyslipidemia in patients on chronic hemodialysis]. AB - Disorder of blood lipids plays an important role in atherosclerosis progress in patients ongoing chronic haemodialysis (PCHD). These patients have specific features of blood lipids with increment of triglycerides and decrement of HDL cholesterol. Phenotype of lipid disorder in PCHD is mostly type IV according to Fredrickson (30%), and IIA and IIB fenotypes are less frequent. About 9% of lipid disorders in PCHD are isolated increase of Lp(a). Main reason of hypertriglyceridemia in PCHD is attenuated metabolism of VLDL-cholesterol because of lipoprotein lipasis inhibition. There are changes in lipoproteins quality, specially changes in LDL particle have atherogenic potential. Renal dyslipidemia treatment must be vigorous in the early stages of renal insufficiency. Treatment can be dietary measures (specially omega-3-fatty acids), statins, gemfibrozil, intravenous L-carnitin and bicarbonate given per os. Haemodialysis modifications such as highflux haemodialysis, low molecular weight heparin, vitamin E coated dialyzers and LDL-apheresis in extreme cases have important role in renal dyslipidemia treatment. PMID- 12899099 TI - [Allergic disorders of the respiratory system and skin as a public health problem]. AB - The prevalence of allergic diseases is rapidly increasing over the last 40 years, especially in most industrialized countries of the world. Manifestations of allergic diseases are observed in about 35% of the general population. In this paper, the most frequent allergic respiratory and skin diseases are described (rhinitis, asthma, atopic eczema/dermatitis syndrome, contact eczema/dermatitis, urticaria). The most important individual and environmental factors involved in the occurrence of allergic diseases are described: genetic predisposition, exposure to allergens, environmental pollution and modern life style are discussed. The results of our investigations related to the prevalence of allergy markers in general and working population are presented. The preventive measures concerning allergic diseases are considered. PMID- 12899100 TI - [Modern approach to hospital care of children]. AB - The article presents children's hospitalization in the Republic of Croatia and in the world, with the accent on contemporary awareness and efforts in humanization of children's hospitalization. First of all, that means shorter children's hospitalization; pledge on the rights of parent's cooperation in medical care and all treatments; reduction of physical and emotional pain of the children, and also other different contents for children in hospital. Such approach to the children in hospital requires the interdisciplinary approach, with participation of all experts in sick children and their family care. PMID- 12899101 TI - Differences in worldwide tobacco use by gender: findings from the Global Youth Tobacco Survey. AB - The World Health Organization (WHO) attributes 4.9 million deaths annually to tobacco. That figure could reach 10 million by 2030. The Global Youth Tobacco Survey (GYTS), an international surveillance project developed jointly by WHO and the US Centers for Disease Control and Prevention (CDC), enables countries to monitor youth tobacco use and guide implementation and evaluation of tobacco prevention and control programs. The GYTS has been completed at 121 sites in 76 countries plus the Gaza Strip/West Bank, with national-level data generated in 52 countries, and city, state, or provincial/regional data generated in 24 countries. This paper reports on gender differences in tobacco use among young people in the six WHO Regions worldwide. Two unexpected findings emerged from the study. First, little difference existed between the genders in cigarette smoking or in use of other tobacco products. From 120 sites that collected data on cigarette smoking by boys and girls, more than one-half (n = 61) showed no difference by gender. For other tobacco products, 82 of 117 sites (70.1%) showed no difference by gender. Second, analysis revealed surprisingly high use of other tobacco products compared to cigarette smoking. Findings suggest programs should focus broadly on all tobacco products, not just cigarettes. Also, programs need gender-sensitive components that focus on unique consequences for females, such as effects on reproduction. Lack of gender differences in the study underscores the potential growth of the tobacco epidemic, especially among women in developing countries--where most sites in this study were located. PMID- 12899102 TI - Children with diabetes: perceptions of supports for self-management at school. AB - Diabetes affects many children. Researchers know little about children's perceptions of what type of support they need at school, which was a focus of this study. Group interviews and surveys examined children's perceptions of support in caring for their diabetes (type I diabetes) from school nurses, teachers, and friends. Results indicated the children felt supported at school, but improved flexibility by teachers and nurses (e.g., let me keep my meter with me always) and individualized care plans may improve their ability to manage their diabetes at school. Participating in after-school activities may be difficult for middle school youth. Children reported they needed additional help and support to cope with hypoglycemic episodes. PMID- 12899103 TI - Integrating school-based and therapeutic conflict management models at schools. AB - Including children with emotional and behavioral needs in mainstream school systems leads to growing concern about the increasing number of violent and nonviolent conflicts. Schools must adapt to this evolution and adopt a more therapeutic dimension. This paper explores the possibility of integrating school based and therapeutic conflict management models and compares two management models: a school-based conflict management program. Teaching Students To Be Peacemakers; and a therapeutic conflict management program, Life Space Crisis Intervention. The authors conclude that integration might be possible, but depends on establishing a positive school atmosphere, the central position of the teacher, and collaborative and social learning for pupils. Further implementation of integrated conflict management models can be considered but must be underpinned by appropriate scientific research. PMID- 12899104 TI - Association of age at onset of cigarette and alcohol use with subsequent smoking and drinking patterns among Japanese high school students. AB - This study examined relationships between age of smoking and drinking initiation, and subsequent patterns of smoking and drinking among Japanese high school students. Self-administered anonymous questionnaires were conducted using a sample of 1,466 students in grades 10 through 12 at seven public senior high schools in urban areas of Okinawa, Japan. After controlling for demographic variables, students who began smoking at age 12 or younger were more likely to become heavy smokers and heavy drinkers than those who began smoking at ages 15 or older. Age of smoking initiation was not related to risks for current smoking and current drinking. Students who started drinking at age 12 or younger, and at 13 or 14 years, were significantly associated with current smoking, current drinking, heavy smoking, and heavy drinking, compared to students who began drinking at ages 15 or older. Early onset smoking and drinking represent significant predictors for subsequent use of cigarettes and alcohol. These findings suggest that smoking and drinking prevention programs should begin during elementary school. PMID- 12899105 TI - Developing a school functioning index for middle schools. AB - Despite widespread recognition of schools' role in the healthy development of youth, surprisingly little research has examined the relationships between schools' overall functioning and the health-related behavior of students. School functioning could become an important predictor of students' health-related behavior and may be amenable to intervention. This paper describes the development and testing of the School Functioning Index (SFI) as a first step in investigating this question. The index was developed for use with middle schools and conceived as a predictor of students' violent behavior, with the potential for extending research applications to additional health and social behaviors. Using social cognitive theory, social ecological theory, and social disorganization theory as guides, three domains were identified to operationalize school functioning and identify candidate SFI items: 1) resources available to the school and students; 2) stability of the school population; and 3) the schools' performance as a socializing agent for students. Data for candidate SFI items were collected from public archives and directly from 16 middle schools participating in a school-based dietary intervention study. Data collection from schools, particularly concerning student aggressive behavior and disciplinary actions, presented challenges. The final SFI comprised nine items and demonstrated good internal consistency and variability. The SFI was modestly correlated in expected directions with violence and other health behaviors. This work supports the feasibility of combining multiple school-level indicators to create a measure of overall school functioning. Further investigation of validity and more acceptable data collection methods are warranted. PMID- 12899106 TI - Using health infomercials to develop media literacy skills. PMID- 12899107 TI - Dental disadvantage among the disadvantaged: double jeopardy for rural school children. PMID- 12899108 TI - Dyslipidaemia and cardiovascular disease risk associated with antiretroviral drugs. PMID- 12899109 TI - Indinavir did not further increase mean triglyceride levels in HIV-infected patients treated with nucleoside reverse transcriptase inhibitors: an analysis of three randomized clinical trials. AB - OBJECTIVES: Metabolic abnormalities including hyperlipidemia have developed in patients infected with the human immunodeficiency virus (HIV) after treatment with protease inhibitor drugs. It is unclear whether the deleterious effects on plasma triglyceride concentrations observed in patients receiving highly active antiretroviral therapy are a class effect of protease inhibitors. Hypertriglyceridemia may constitute a risk factor for cardiovascular disease. The purpose of this retrospective analysis of HIV-infected patients enrolled in three randomized, double-blind trials of indinavir therapy was to determine whether indinavir use was associated with a larger increase in triglyceride levels than treatment without a protease inhibitor. METHODS: Using a mixed-effects model, we compared average changes in nonfasting plasma triglyceride levels among randomized treatment groups for each protocol separately. RESULTS: The median increase in triglyceride levels during the 1st year of antiretroviral monotherapy was less with indinavir than with either zidovudine or stavudine. The combination of indinavir and nucleoside-analogue reverse-transcriptase inhibitors (NRTI) resulted in smaller increments in triglyceride levels than NRTI monotherapy. Indinavir also augmented the reduction in triglyceride levels observed with combination therapy using zidovudine and lamivudine in persons with far advanced HIV-infection. However, up to 7% of patients receiving a NRTI and indinavir experienced elevations of nonfasting triglyceride levels in excess of 750 mg/dl. CONCLUSIONS: On average, the combination of indinavir and NRTI therapy was not associated with a greater elevation of non-fasting triglyceride levels in HIV infected men with at least moderately advanced immunosuppression than treatment with NRTI drugs alone. PMID- 12899110 TI - Use of the New Zealand Intensive Medicines Monitoring Programme to study the levonorgestrel-releasing intrauterine device (Mirena). AB - PURPOSE: To demonstrate how the Intensive Medicines Monitoring Programme (IMMP) can be used to monitor adverse events associated with an intrauterine device, using the levonorgestrel-releasing intrauterine device (Mirena) as an example. METHODS: A long-term prospective observational cohort study using Prescription Event Monitoring (PEM) is currently being undertaken in women using Mirena in New Zealand. This report describes the method used and reports the early results for those women who used the device between March 1998 and March 2001. Adverse events were recorded by inserting doctors and general practitioners on registration forms and systematic follow-up questionnaires. RESULTS: Between March 1998 and March 2001, the IMMP received 3519 registration forms for insertions in 3452 women. 'Difficult insertion' was the most frequently reported event (3.6% of all insertions). Approximately, 2% of the Mirena insertions were carried out under GA and there were three uterine perforations (0.9 per 1000 insertions) in the total cohort. To date, follow-up questionnaires have been processed for 495 patients. The response rate for these was 83%. CONCLUSION: As adapted in the IMMP, PEM is an effective tool for the early post-marketing surveillance of an intrauterine device in real life clinical practice. PMID- 12899111 TI - Prescribing patterns of tricyclic and selective serotonin reuptake inhibitor antidepressants among a sample of adolescents and young adults. AB - PURPOSE: There are few studies that document antidepressant prescribing in young patients. Although tricyclic antidepressants (TCAs) are considered equal in efficacy to selective serotonin reuptake inhibitors (SSRIs), the latter have an improved side effect profile. The aim of this study was to investigate the prescribing patterns of TCAs and SSRIs among adolescents and young adults, with reference to prescribing frequency, cost and dose. METHOD: A retrospective drug utilization study was conducted over a 14-month period, from January 2000 to February 2001. RESULTS: There were 166 antidepressants prescribed to 98 adolescents and young adults. TCAs were prescribed more frequently than SSRIs, with amitryptiline and fluoxetine being the two most frequently prescribed antidepressants. Fluoxetine accounted for a higher ratio of cost to prescribing frequency than amitryptiline. Amitryptiline was issued in small quantities of tablets, resulting in a low average calculated prescribed daily dose (PDD). Duration of treatment was not considered optimal for SSRIs or TCAs. CONCLUSION: This study elicits prescribing patterns that contribute to the relative scarcity of data on antidepressant drugs for young patients. PMID- 12899112 TI - Factors influencing prescription of CNS medications in different elderly populations. AB - Factors influencing the use of neuroleptic and other CNS-acting medications ('antidepressants' and 'hypnotics/anxiolytics') were examined in different elderly populations (> 65 years): long-stay care hospital subjects (n = 381), nursing home subjects (n = 1247), private residential home subjects (n = 321), statutory residential home subjects (n = 525), old age psychiatry facility subjects (n = 48), community dwellers (n = 97), in different geographical areas (urban n = 1223 and rural n = 1396). Neuroleptics were prescribed in 28% of the individuals, hypnotics/anxiolytics in 33% and antidepressants in 12%. Prescription of neuroleptics was significantly predicted by institutional placement (other than long-stay care hospital facility), lower cognitive function and rural geographical area. Age greater than 75 years and lower functional score (lower dependency) significantly decreased the likelihood of receipt of neuroleptics. Antidepressant use was significantly less likely in male subjects, patients with cognitive impairment and patients greater than 85 years. Institutional placement (other than long-stay care hospital facility and old age psychiatry facility) and rural locality predicted increased likelihood of antidepressant prescription. Institutional placement (other than long-stay care hospital facility) and geographical (rural) location were significant independent predictors of hypnotic/anxiolytic use. Prescription of hypnotic/anxiolytic drug class was significantly less likely in those individuals with lower cognitive status. In this representative elderly cohort, patient characteristics: age, sex, cognitive score, functional score, place of residence and geographical location markedly influenced drug utilisation. PMID- 12899113 TI - Medication and fitness to drive. AB - PURPOSE: The aim of this study is to analyze the consumption patterns of medicaments among motor vehicle drivers who attend 'Medical Driving Test Centres' and the relation between habitual consumption of medicaments and fitness to drive. METHODS: The study was carried out on 8043 drivers who attended 25 Medical Driving Test Centres. RESULTS: 24.7% of drivers chronically consume medicaments while 6.8% consume medicaments along with alcohol every day. Of those who chronically consume medicaments with a warning about the medications on driving, 65.8% were considered 'fit' to drive, 27.3% 'fit with restrictions', 5.1% 'suspended' and 0.4% 'unfit'. CONCLUSIONS: The results show how frequent the consumption of medicaments along with alcohol is and that the great majority of drivers who take medicaments are considered fit to drive. PMID- 12899114 TI - The adjunctive use of a centralised database in the monitoring of clozapine related neutropenia. AB - OBJECTIVE: To assess the effectiveness of centralised routine monitoring of blood counts in subjects receiving clozapine. DESIGN: A cohort study based on a prospective drug exposure database. SETTING AND PARTICIPANTS: Patients with schizophrenia in 38 specialised centres across Australia. OUTCOME MEASURES: The time from detection of neutropenia to the interruption of therapy, the time to recovery of the absolute neutrophil count, the eventual outcome of subjects developing neutropenia. RESULTS: A total of 1500 subjects were registered on the database. Seven subjects developed severe neutropenia (a neutrophil count < 1.5 x 10(9)/L) whilst on therapy. Mean time to cessation of therapy was 1.6 days (range < 1-6 days). Neutrophil counts recovered to normal levels in all subjects after 6.4 days (range < 1-13). CONCLUSIONS: In the population investigated, the addition of a centralised database to clinical monitoring resulted in very rapid detection of severe neutropenia and an equally rapid cessation of therapy. In the sample investigated, this led to very low morbidity and zero mortality. PMID- 12899115 TI - Use of psychiatric drugs in Slovenia in comparison to Scandinavian countries. AB - PURPOSE: To find out the consumption of psychotropic drugs in Slovenia and the extent to which Slovenian consumption correlates to Scandinavian countries. METHODS: Data of ambulatory prescribing of psychiatric drugs in Slovenia for the years 1999 and 2000 and data for Norway, Sweden and Denmark are shown. All data are expressed in defined daily doses per thousand inhabitants per day (DDD's). RESULTS: The use of all neuroleptics in Slovenia is lower (5.3 vs 6.1-7.6 DDD's), however the share of atypicals (34%) is comparable to Scandinavian countries. Use of anticholinergics is up to five times greater (1.6 vs 0.3-0.5 DDD's). The use of lithium is five time lower (0.2 vs 1.1-1.4). Regarding the use of antidepressants, Slovenia is behind Scandinavia (12.2 vs 30-47 DDD's), whereas the share of SSRI's (73%) is at the same level. The use of anxiolytics is higher (25.4 vs 16-22.4 DDD's) and ratio with antidepressants is also unfavourable. The use of hypnotics and sedatives is favourably low (10.7 vs 31.8-44.6 DDD's). CONCLUSIONS: Low use of hypnotics and sedatives is favourable. The share of new neuroleptics and antidepressants is comparable. The use of antidepressants is probably too low. However, prescribing guidelines for anxiolytics, anticholinergics (too high use of both group) and lithium (too low use) should be introduced in Slovenia. PMID- 12899116 TI - The Cuban experience in focusing pharmaceuticals policy to health population needs: initial results of the National Pharmacoepidemiology Network (1996-2001). AB - In 1996, a National Pharmacoepidemiology Network was set up in Cuba in order to disseminate problem-oriented information and develop continuing education activities on drug therapy, to perform research on drug utilisation and to promote interventions to improve drug use. It is coordinated by the Centre for the Development of Pharmacoepidemiology, which is also in charge of regulatory and administrative duties and collaborates with the National Pharmaceutical Industry. By 2001, a number of educational and information initiatives had been undertaken, particularly in primary health care, which resulted in improvement of indicators of drug use. Cuba keeps good health standards in spite of a short health budget. PMID- 12899117 TI - Validation studies of drug exposures in pregnant women. AB - The aim of these studies was to check the quality of data collection concerning drug use during pregnancy in the Hungarian Case-Control Surveillance of Congenital Abnormalities from four different aspects. (a) The parents of non respondent cases were visited at home to collect data, but the Ethics Committee did not allow collection of comparable data from non-respondent control women. Thus only 200 selected non-respondent controls were visited at home and questioned about drug use as part of a validation study. There was no difference in the distribution and occurrence of frequently used drugs between the respondent and non-respondent control groups. (b) Data collection was compared between the usual method of the Case-Control Surveillance and personal interview in case with congenital limb deficiencies. In general, the two methods produced similar results with respect to drug use, but the personal interview revealed a higher use of unintended contraceptive pills while the use of antenatal care logbooks in the Case-Control Surveillance showed a higher use of drugs to prevent threatened abortion and preterm delivery. (c) A check of source and completeness of drug exposure showed that about one-quarter of the drugs used during pregnancy were not recorded in the Case-Control Surveillance, and the proportion of lack of recall was about 23%. A small group of women (2.4%) did not use prescribed drugs due to the suspected teratogenic risk. (d) The teratogenic labelling of drugs deters pregnant patients from the use of prescribed drugs, or at least shortens the duration of drug intake. Thus the evaluation of drug use on the basis of medically recorded prescribed drugs and/or usual recommendations produces a bias thus it is necessary to complete it with information from the patients. PMID- 12899118 TI - Doctors' prescription behaviour regarding dosage recommendations for preparations of kava extracts. AB - BACKGROUND: There are many publications on the use of kava extracts as a treatment alternative to tricyclic antidepressants and benzodiazepines, but little has been done to investigate the potential adverse effects that may be associated with prolonged or high-dose usage. OBJECTIVES: To investigate the extent to which physicians comply with the recommended daily dose (RDD) of an antidepressant such as kava and the consequences of this behaviour with regard to the safety and efficacy of frequently prescribed kava extracts (Trade names: Antares, Laitan and Kavasporal forte). METHOD: The analysis was carried out using the patient database MediPlus, which provides anonymous access to a representative and valid panel of physicians and patients in Germany. RESULTS: Prescriptions for Antares showed that the RDD of 1 tablet per day was exceeded in 58.2% of the cases, while those for Laitan showed that the RDD of 1 capsule per day was significantly exceeded in 75.6% of the cases; a one-off prescription was issued in 66% of the cases. As a result, the required effect diminished very quickly. Prescriptions for Kavasporal forte showed that the daily dosage of 2 capsules per day complied with the recommendations in only 49.1% of the cases; in 38.8% of the cases, the dosage dropped below the respective recommendation. As a consequence, the lower-dosed Kavasporal forte was prescribed for longer periods than the higher-dosed Antares and Laitan. CONCLUSIONS: In the case of a low dosage recommendation, we saw a trend to over-dose in prescribing behaviour, which increases the risk of undesirable adverse reactions. On the other hand, we saw a trend to under-dose in the case of a higher dosage recommendation. This fact may explain the unexpected inefficacy of the therapy. These trends affected the duration of therapy, whereby a dosage lower than the RDD was prescribed for a longer period than a dosage higher than the RDD. PMID- 12899119 TI - Increased suicide attempt rate among patients interrupting use of atypical antipsychotics. AB - PURPOSE: The aim of the study is to investigate whether patients at risk to commit suicide could be identified based on their drug refill patterns. METHODS: Data for this study were obtained from the PHARMO system comprising drug dispensing data linked to hospital discharge records. All patients aged 15-45 years, suspected to suffer from schizophrenia and exclusive users of olanzapine or risperidone were selected. The use of antipsychotics was converted into treatment episodes of uninterrupted use. Therapy non-compliance was defined as a drug holiday of at least 30 days. The follow-up started with a treatment episode of at least 90 days. Relative risks for suicide attempts and 95% confidence limits due to drug holidays were estimated using Poisson regression analyses. RESULTS: Of 603 patients, 33% interrupted treatment for at least 30 days. An increased suicide attempt rate was observed when comparing uninterrupted and interrupted drug use (20.0/1000 person years vs 72.1/1000 person years, respectively). A four-fold increased risk for attempting suicide among patients with drug holidays was found (RR adjusted for age and gender 4.2, 95% CI: 1.7 10.1) compared to patients without drug holidays. CONCLUSIONS: Patients who do not refill atypical antipsychotics in time can be identified in the pharmacy and are most likely those with an increased risk to commit suicide. PMID- 12899120 TI - Postmarketing drug dosage changes. PMID- 12899121 TI - Understanding drug-herb interactions. PMID- 12899122 TI - Current awareness in pharmacoepidemiology and drug safety. PMID- 12899123 TI - Monopyrrolic natural compounds including tetramic acid derivatives. PMID- 12899124 TI - [Endometriosis and diferelin]. PMID- 12899125 TI - [Toxicity of stannum and its determination]. PMID- 12899126 TI - EMS takes flights. PMID- 12899127 TI - Mortality trend in female breast cancer in Japan: 1960-2000. PMID- 12899128 TI - [Preclinical evaluation of drug induced QT prolongation]. PMID- 12899129 TI - Radiation dose in helical CT for detection of pulmonary embolism. PMID- 12899130 TI - Contrast ultrasound of the urethra in children. PMID- 12899131 TI - [To carry out the vitreoretinal surgery energetically and steadily]. PMID- 12899132 TI - [Bronchoalveolar carcinoma]. PMID- 12899133 TI - Emergence and global spread of a dengue serotype 3, subtype III virus. AB - Over the past two decades, dengue virus serotype 3 (DENV-3) has caused unexpected epidemics of dengue hemorrhagic fever (DHF) in Sri Lanka, East Africa, and Latin America. We used a phylogenetic approach to evaluate the roles of virus evolution and transport in the emergence of these outbreaks. Isolates from these geographically distant epidemics are closely related and belong to DENV-3, subtype III, which originated in the Indian subcontinent. The emergence of DHF in Sri Lanka in 1989 correlated with the appearance there of a new DENV-3, subtype III variant. This variant likely spread from the Indian subcontinent into Africa in the 1980s and from Africa into Latin America in the mid-1990s. DENV-3, subtype III isolates from mild and severe disease outbreaks formed genetically distinct groups, which suggests a role for viral genetics in DHF. PMID- 12899134 TI - A case of hemiparesis associated with cranial nerve lesions due to intravascular lymphomatosis(2003:4b). AB - Intravascular lymphomatosis is a rare variant of non-Hodgkin's lymphoma with an unusual predilection for the central nervous system. Most cases are not diagnosed until postmortem because of variable clinical presentation and non-specific laboratory findings. We studied a 41-year-old lady who presented with progressive neurological symptoms. MR showed multifocal grey and white matter lesions. Brain biopsy showed an intravascular lymphoma of the B-cell lineage. This report illustrates the ischaemic origin of the radiological lesions, as they are all hyperintense on the diffusion-weighted images. This has, to our knowledge, never been published before. MR did not show any enhancement after intravenous Gadolinium-DTPA (parenchymal or meningeal)which is a very uncommon finding in this entity. Intravascular lymphomatosis should be taken into account in the differential diagnosis of repeated cerebral ischemia of unclear aetiology. PMID- 12899135 TI - [84th Roentgen Congress 28 -31 May 2003 in retrospect]. PMID- 12899136 TI - [Pay attention to the continuative medical education to elevate the quality of professionals in ophthalmology]. PMID- 12899137 TI - [Malpractice judgment: content of referral determines extent of the examination]. PMID- 12899138 TI - Taenia solium Cysticercosis, Irian Jaya, Indonesia. PMID- 12899139 TI - Recombinant vaccine-derived poliovirus in Madagascar. PMID- 12899140 TI - West Nile virus infection in crocodiles. PMID- 12899141 TI - Rickettsia aeschlimannii in Spain: molecular evidence in Hyalomma marginatum and five other tick species that feed on humans. PMID- 12899142 TI - [To further elevate diagnostic and therapeutic levels of retinal detachment in our country]. PMID- 12899143 TI - Hantaviruses in Sao Paulo State, Brazil. PMID- 12899144 TI - Israeli spotted fever Rickettsia in Sicilian Rhipicephalus sanguineus ticks. PMID- 12899145 TI - Co-feeding transmission and its contribution to the perpetuation of the Lyme disease spirochete Borrelia afzelii. PMID- 12899146 TI - West Nile virus southeast conference. PMID- 12899147 TI - West Nile virus and wildlife health. PMID- 12899148 TI - S-adenosyl-L-methionine for treatment of depression, osteoarthritis, and liver disease. PMID- 12899149 TI - Understanding and Managing Sexually Coercive Behavior. June 7-9, 2002. Washington, DC, USA. Proceedings. PMID- 12899150 TI - [Combined treatment for infectious keratopathy]. PMID- 12899151 TI - Brain03 and BrainPET '03. Abstracts of the XXIst International Symposium on Cerebral Blood Flow, Metabolism and Function and the VIth International Conference on Quantification of Brain Function with PET. June 29-July 3, 2003. Calgary, Alberta, Canada. PMID- 12899152 TI - Proceedings of the 7th International Conference on Nerve Growth Factor, dedicated to Rita Levi-Montalcini. Modena, Italy, 15-19 May 2002. PMID- 12899153 TI - [Devote attention to regular training, and spread phacoemulsification for cataract extraction energetically and safely]. PMID- 12899154 TI - And God created the Heavens and the Earth. PMID- 12899155 TI - Emergency contraception and risk of ectopic pregnancy: is there need for extra vigilance? PMID- 12899156 TI - The emergency medicine workforce. PMID- 12899157 TI - Caveat emptor: the positive D-dimer slippery slope. PMID- 12899159 TI - Deadly game among children and adolescents. PMID- 12899160 TI - Proceedings of the 6th Symposium on Instrumental Analysis. Graz, Austria, 24-27 June 2001. PMID- 12899161 TI - Allelochemical interaction between plant and other organisms--combined approach of laboratory and field. AB - This issue of Biological Sciences in Space is dedicated to "Allelochemical interaction between plant and other organisms-Combined approach of laboratory and field". Papers were presented at the symposium that was held in Osaka on March 27, 2003. PMID- 12899162 TI - [Improve the quality of glaucoma surgery]. PMID- 12899163 TI - [New option for successful therapy of chronic hepatitis B]. PMID- 12899164 TI - [Survival rate of HIV infected patients depends on CD4 count at therapy initiation]. PMID- 12899165 TI - [HIV infections: further options for combination therapy]. PMID- 12899166 TI - [Gambian giant pouched rat and prairie dogs: monkeypox outbreak in America]. PMID- 12899167 TI - [Proceedings of the 3rd Glaucoma Congress of the Polish Society of Ophthalmology. October 10-12, 2002]. PMID- 12899168 TI - Cases from the Osler Medical Service at Johns Hopkins University. Wegener's granulomatosis. PMID- 12899170 TI - Medical forecasting in classic antiquity and now, the same or different? PMID- 12899169 TI - Robert Burns: an Ayrshire lad. PMID- 12899172 TI - A Scottish women's hospital on the western front. PMID- 12899171 TI - Professor Macleod's researches on the source of insulin. PMID- 12899173 TI - William Smellie: a conservative in labor. PMID- 12899174 TI - Not least in the crusade: the centenary of the Royal Army Medical Corps. PMID- 12899175 TI - Architecture for hope: some modernist aspects of design and architecture and their association with tuberculosis. PMID- 12899176 TI - Murder in the university. PMID- 12899177 TI - [Study of a patient's right protection system: modeled after the patient's right protection system of the state of NY]. PMID- 12899178 TI - The canine veterinary medicine in the Middle Ages according to the Livre de chasse by Gaston Phoebus. AB - Gaston Phoebus (1331-1391) of Foix-Bearn was a mighty vassal of the king of France as well as of the king of England. His book, the Livre de Chasse was the first technical treaty of this scope. It was fundamental for the regulation of hunting in Europe, as it was copied and plagiarized later on by a number of authors. Chapter 16 is about diseases of the dog and their treatment. In this article Phoebus views about pathology, etiology and therapy are discussed. The absence of superstition, magic and preconceived notions is remarkable. His writings are based on personal observations which are detailed. His therapy is generally rational and probably often effective, never harmful. His love of hounds is evident. It is regrettable that this highly rational approach didn't have a longer lasting effect on the development of canine medicine. PMID- 12899179 TI - "Death talk": debating euthanasia and physician-assisted suicide in Australia. PMID- 12899180 TI - "Death talk": debating euthanasia and physician-assisted suicide in Australia. PMID- 12899181 TI - "Death talk": debating euthanasia and physician-assisted suicide in Australia. PMID- 12899182 TI - "Death talk": debating euthanasia and physician-assisted suicide in Australia. PMID- 12899183 TI - "Death talk": debating euthanasia and physician-assisted suicide in Australia. PMID- 12899184 TI - Deficient response modulation and emotion processing in low-anxious Caucasian psychopathic offenders: results from a lexical decision task. AB - The clinical and research literatures on psychopathy have identified an emotion paradox: Psychopaths display normal appraisal but impaired use of emotion cues. Using R. D. Hare's (1991) Psychopathy Checklist-Revised and the G. S. Welsh Anxiety Scale (1956), the authors identified low-anxious psychopaths and controls and examined predictions concerning their performance on a lexical-decision task. Results supported all the predictions: (a) low-anxious psychopaths appraised emotion cues as well as controls; (b) their lexical decisions were relatively unaffected by emotion cues; (c) their lexical decisions were relatively unaffected by affectively neutral word-frequency cues; and (d) their performance deficits were specific to conditions involving right-handed responses. The authors propose that deficient response modulation may underlie both the emotional and cognitive deficits associated with low-anxious psychopaths. PMID- 12899185 TI - Eye movement assessment of emotional processing in anxiety. AB - Eye fixations of participants high or low in trait anxiety were monitored during reading of context sentences predicting threatening or nonthreat events, followed by sentences in which a target word represented the predictable event or an inconsistent event. No effects were found on the target word but on the regions following it. When this word represented a threatening event suggested by the context, high anxiety facilitated reading of the posttarget region. When the target word was inconsistent with a threatening event, high anxiety was associated with interference in the final region. No such effects occurred with nonthreat sentences. This reveals selective prediction of threat in high anxiety. However, rather than being automatic, this bias involves elaboration that takes time to develop. PMID- 12899186 TI - Effects of normal aging and Alzheimer's disease on emotional memory. AB - Recall is typically better for emotional than for neutral stimuli. This enhancement is believed to rely on limbic regions. Memory is also better for neutral stimuli embedded in an emotional context. The neural substrate supporting this effect has not been thoroughly investigated but may include frontal lobe, as well as limbic circuits. Alzheimer's disease (AD) results in atrophy of limbic structures, whereas normal aging relatively spares limbic regions but affects prefrontal areas. The authors hypothesized that AD would reduce all enhancement effects, whereas aging would disproportionately affect enhancement based on emotional context. The results confirmed the authors' hypotheses: Young and older adults, but not AD patients, showed better memory for emotional versus neutral pictures and words. Older adults and AD patients showed no benefit from emotional context, whereas young adults remembered more items embedded in an emotional versus neutral context. PMID- 12899187 TI - Sadness and amusement reactivity differentially predict concurrent and prospective functioning in major depressive disorder. AB - Depressed individuals often fail to react to emotionally significant stimuli. The significance of this pattern of emotional dysregulation in depression is poorly understood. In the present study, depressed and nondepressed participants viewed standardized neutral, sad, fear, and amusing films; and experiential, behavioral, and physiological responses to each film were assessed. Compared with nondepressed controls, depressed participants reported sadness and amusement in a flattened, context-insensitive manner. Those depressed participants who reported the least reactivity to the sad film exhibited the greatest concurrent impairment. Prospectively, the depressed participant who exhibited the least behavioral and heart rate reactivity to the amusing film were the least likely to recover from depression. Loss of the context-appropriate modulation of emotion in depression may reflect a core feature of emotion dysregulation in this disorder. PMID- 12899188 TI - Comparison of emotional responses in monkeys with rhinal cortex or amygdala lesions. AB - Four emotionally arousing stimuli were used to probe the behavior of monkeys with bilateral ablations of the entorhinal and perirhinal cortex. The monkeys' behavioral changes were then contrasted with those observed earlier (M. Meunier, J. Bachevalier, E. A. Murray, L. Malkova, & M. Mishkin, 1999) in monkeys with either neurotoxic or aspiration lesions of the neighboring amygdala. Rhinal cortex ablations yielded several subtle behavioral changes but none of them resembled any of the disorders typically seen after amygdalectomies. The changes produced by rhinal damage took mainly the form of heightened defensiveness and attenuated submission and approach responses; that is, just the opposite of some of the most distinctive symptoms following amygdala damage. These findings raise the possibility that the rhinal cortex and amygdala have distinct, interactive functions in normal behavioral adaptation to affective stimuli. PMID- 12899189 TI - Individual differences in misremembering pre-psychotherapy distress: personality and memory distortion. AB - In 2 studies, clients were asked, after terminating psychotherapy, to complete a measure of symptomatic distress exactly as they had in their pre-therapy assessment. Most clients overestimated their pre-therapy distress, which may lead to an illusion of positive change. A 3rd study found no overestimation of previously reported distress in a control sample. The degree of overestimation for psychotherapy clients was positively correlated with anxiety, depression, and neuroticism, and negatively correlated with ego strength, self-deception, and lying. Distress levels at termination mediated these effects of individual differences. Individual differences were also correlated with specific types of recall error, such as adding to, exaggerating, omitting, and minimizing previously reported symptoms. Misremembering prior emotions may both reflect and create individual differences. PMID- 12899190 TI - Observing emotion in infants: facial expression, body behavior, and rater judgments of responses to an expectancy-violating event. AB - Eleven-month-old European-American, Japanese, and Chinese infants (ns = 23, 21, and 15, respectively) were videotaped during baseline and stimulus episodes of a covert toy-switch procedure. Infants looked longer at the object during the expectancy-violating event (stimulus episode) but did not produce more surprise related facial expressions. American and Japanese infants produced more bodily stilling during stimulus than baseline, and American infants also produced more facial sobering. Naive raters viewing both episodes could correctly identify the expectancy-violating event. Rater judgments of surprise were significantly related to infants' bodily stilling and facial sobering. Judgments of interest were related to cessation of fussing. Thus, observer judgments of infant emotions can be systematically related to behaviors other than prototypic emotional facial expressions. PMID- 12899191 TI - Linkages between facial expressions of anger and transient myocardial ischemia in men with coronary artery disease. AB - The authors examined whether facial expressions of emotion would predict changes in heart function. One hundred fifteen male patients with coronary artery disease underwent the Type A Structured Interview, during which time measures of transient myocardial ischemia (wall motion abnormality and left ventricular ejection fraction) were obtained. Facial behavior exhibited during the ischemia measurement period was videotaped and later coded by using the Facial Action Coding System (P. Ekman & W. V. Friesen, 1978). Those participants who exhibited ischemia showed significantly more anger expressions and nonenjoyment smiles than nonischemics. Cook-Medley Hostility scores did not vary with ischemic status. The findings have implications for understanding how anger and hostility differentially influence coronary heart disease risk. PMID- 12899192 TI - The relations of parental emotional expressivity with quality of Indonesian children's social functioning. AB - In Western societies, parental expression of positive emotion has been positively related to the quality of children's social functioning, whereas their expression of negative emotion has been negatively or inconsistently related. The relations of parental expressivity to 3rd-grade Indonesian children's dispositional regulation, socially appropriate behavior, popularity, and sympathy were examined. Parents, teachers, and peers reported on children's social functioning and regulation, and parents (mostly mothers) reported on their own expression of emotion in the family. Generally, parental expression of negative emotion was negatively related to the quality of children's social functioning, and regression analyses indicated that the relations of parental negative expressivity to children's popularity and externalizing behaviors might be indirect through their effects on children's regulation. Unexpectedly, parental expression of positive emotion was unrelated to children's social functioning. PMID- 12899193 TI - Cortical brain regions engaged by masked emotional faces in adolescents and adults: an fMRI study. AB - Face-emotion processing has shown signs of developmental change during adolescence. Functional magnetic resonance imaging (fMRI) was used on 10 adolescents and 10 adults to contrast brain regions engaged by a masked emotional face task (viewing a fixation cross and a series of masked happy and masked fearful faces), while blood oxygen level dependent signal was monitored by a 1.5 T MRI scanner. Brain regions differentially engaged in the 2 age groups were mapped by using statistical parametric mapping. Summed across groups, the contrast of masked face versus fixation-cross viewing generated activations in occipital-temporal regions previously activated in passive face-viewing tasks. Adolescents showed higher maxima for activations in posterior association cortex for 3 of the 4 statistical contrasts. Adolescents and adults differed in the degree to which posterior hemisphere brain areas were engaged by viewing masked facial displays of emotion. PMID- 12899194 TI - Offset control during recurrent 20% carbon dioxide-enriched air induction: relation to individual difference variables. AB - Although control over aversive events maintains a central role in contemporary models of anxiety pathology, particularly panic disorder, there is little understanding about the emotional consequences of specific types of control processes. In the present study, offset control over 8 20% carbon dioxide enriched air administrations was experimentally manipulated in a large nonclinical population (n = 96) varying in anxiety sensitivity (high or low) and gender. Dependent measures included self-reported anxiety, affective reports of valence, arousal, emotional control, and physiological indices of heart rate and skin conductance. High anxiety-sensitive participants who lacked offset control reported significantly greater elevations in self-reported anxiety, emotional displeasure, arousal, and dyscontrol relative to their yoked counterparts with offset control. In contrast, low anxiety-sensitive individuals responded with similar levels of cognitive and affective distress regardless of the offset control manipulation. Although the provocation procedure reliably produced bodily arousal relative to baseline, at a physiological level of analysis, no significant differences emerged across conditions. These findings are discussed in relation to offset control during recurrent interoceptive arousal, with implications for better understanding anxiety about abrupt bodily sensations. PMID- 12899195 TI - Anxiety and cognitive inhibition. AB - In 3 experiments, the authors investigated whether anxiety proneness is associated with impaired inhibitory processing. Participants made speeded decisions requiring inhibition of threatening or neutral meanings of ambiguous words, which were inappropriate in their current context. In Experiment 1 there were no differences found in inhibitory processing associated with anxiety. However, in Experiment 2, when the capacity for controlled processing was reduced by imposition of a mental load, anxious individuals showed a response pattern consistent with a general impairment of inhibitory processing. In Experiment 3, a group who had experienced a traumatic event also showed evidence of impaired inhibition, despite the absence of additional load. Thus anxiety proneness is associated with a general deficit of inhibitory processing, but this may be revealed only under conditions that limit the availability of controlled processing resources. PMID- 12899196 TI - Stress affects the selection of relevant from irrelevant stimuli. AB - Two experiments were used to examine the effects of stress on latent inhibition (LI; poorer learning with a previously exposed irrelevant stimulus rather than a novel stimulus). In Experiment 1, stress was induced in college students by threatening participants' self-esteem with a difficult number series completion test that was related to intelligence. In Experiment 2, the participants were job seekers who were either informed or not that the LI test was part of the selection process. In both experiments, LI was attenuated in high- as compared with low-stressed participants. The results suggest that stress and/or anxiety impairs the inhibition of irrelevant-preexposed stimuli. Implications for understanding the impaired selective attentional processes in schizophrenia and schizotypy are discussed. PMID- 12899197 TI - Familial amyotrophic lateral sclerosis and parkinsonism-dementia complex- tauopathy without mutations in the tau gene? AB - We present the clinical and genetic characteristics of a Japanese patient with neuropathologically confirmed familial amyotrophic lateral sclerosis/parkinsonism dementia complex (ALS/PDC). The 68-year-old proband with an 8-year history of parkinsonism and neurogenic amyotrophy and her three siblings suffering from parkinsonism associated with dementia originated from the Kii Peninsula of Japan. The proband's brain exhibited mild frontal lobe atrophy, moderate atrophy of the pes hippocampi, decoloration of the substantia nigra and locus coerules, and atrophy of the anterior root of the spinal cord. Microscopic examinations revealed degeneration of the CA1 portion of the hippocampus to the parahippocampus gyrus, substantia nigra, locus coerules and the spinal anterior horn with Bunina bodies. Neurofibrillary tangles (NFTs) were observed in widespread regions of the central nervous system through the cerebral cortex to the spinal cord. The predominant distribution of NFTs in the the third layer of the cerebral cortex was compatible with the characteristic feature of ALS/PDC in Guam. No tau mutation was found in the proband. The lack of mutations in the tau gene not only in this patient but also in earlier reported cases of ALS in the Western Pacific seems to suggest that other genetic factors may be contributing to ALS/PDC. PMID- 12899198 TI - Apolipoprotein E polymorphism and low density lipoprotein (LDL) oxidation in patients with dementia. AB - In patients with dementia, 29 diagnosed as probably suffering from Alzheimer's disease and 46 subjects with dementia of vascular origin, and in 41 non demented control subjects LDL oxidation in vitro was compared in carriers of various apolipoprotein E alleles. Restriction isotyping was performed by gene amplification and cleavage with Hhal, LDL oxidation was investigated by determination of conjugated dienes and vitamin E (alpha tocopherol) plasma level was measured by HPLC. In subjects with dementia oxidation of LDL was shown to be higher in carriers of epsilon4 allele as compared with non-carriers of this allele. It was especially observed in the propagation phase, which illustrates oxidation intensity after the exhaustion of the antioxidant reserve in LDL. Vitamin E level did not show differences between carriers of different alleles. It is concluded that the differences in oxidation susceptibility of LDL between demented subjects possessing particular apolipoprotein E forms can result partially from differing antioxidant properties of apolipoprotein E isoforms and, in a substantial degree, from the size and quality of LDL. PMID- 12899199 TI - Preclinical development of biodegradable polymer foils for intracerebral delivery of cytotoxic nucleosides. AB - Intracerebral implantation of biodegradable polymers loaded with cytotoxic or radiosensitising nucleoside analogues is a promising treatment strategy for malignant gliomas, which are currently intractable. The aim of the study was to develop biodegradable polymers containing nucleosides which could be implanted intracerebrally. Methods of synthesis were developed for the copolymers composed of D,L-lactide, glycolide and caprolactone in different proportions, as well as a novel method of introducing nucleosides to these copolymers at the polymerisation step. Upon degradation in an aqueous medium some of these copolymers emit nucleosides in micromolar concentration over several months. Their in situ degradation and biocompatibility with brain tissues was assessed by means of scanning and transmission electron microscopy. At the ultrastructural level tissue responses to the copolymer implantation closely resembled the responses to mechanical trauma. PMID- 12899200 TI - Constitutive patterns of RANTES, MCP-1 and MIP-1 alpha expression at the mRNA and protein level during postnatal development of the rat brain. AB - The importance of chemokines seems to extend far beyond their well-known role as mediators of an inflammatory response. The most interesting hypothesis is that these molecules may influence the migration of progenitor cells during development. Primary sensory neurones have been shown to migrate towards RANTES in vitro. Recent evidence has revealed that MCP-1, MIP-1alpha and MIP-1beta are potent chemoattractants for glial cell populations. The exact role of the constitutive appearance of chemokines in the CNS during postnatal development is still largely unknown. The intention was to show whether constitutive temporal and spatial profiles of RANTES, MCP-1 and MIP-1alpha expression vary during the postnatal development of the rat brain. RT-PCR was used to assess the levels of mRNA production at different developmental stages. Semi-quantitative analysis of the immunofluorescence signal from glial cells harbouring chemokines was used to determine the spatial-temporal patterns of protein expression. It has been shown here that all chemokines are constitutive factors within the brain microenvironments where the postnatal migration phenomenon occurs. The chemokines were characterised by variable temporal patterns of mRNA production and distinct spatial-temporal patterns of protein appearance. This may support the differences between RANTES, MCP-1 and MIP-1alpha functional significance in vivo in terms of influence on the migration of distinct cell populations. PMID- 12899201 TI - Expression of macrophage/histiocytic antigens in pleomorphic xanthoastrocytomas. AB - Pleomorphic xanthoastrocytoma (PXA) is a rare variant of a superficial cerebral astrocytoma characterised by distinct clinical and histological features. Its derivation from subpial astrocytes has been proposed, although the capacity of neoplastic cells for expression of different immunohistochemical markers is still under debate. These immunohistochemical studies were performed on eight cases of PXA in order to evaluate the expression and co-expression of glial and macrophage/histiocytic markers in various tumour cell populations. The expression of antigens was examined with the use of single- and double-immunolabelling methods for GFAP, vimentin, LCA, CD68, HLA-class II and MAC 387. All the cases of PXA showed variable immunoreactivity to GFAP, both in spindle-shaped and pleomorphic lipidised tumour cells. A subset of neoplastic cells was stained strongly with HLA-class II monoclonal antibody and with antibody to CD68. The reactivity to LCA and MAC 387 was absent in neoplastic cells, while it was easily evidenced in the non-neoplastic infiltrative component. The immunohistochemical double staining demonstrated the co-expression of GFAP and HLA-class II or CD68 antigens in the cytoplasm of individual neoplastic cells, including large pleomorphic, lipid-laden ones. It seems that tumour cells in PXAs derived from subpial astrocytes reveal monocyte/macrophage immunophenotype and demonstrate the capability of functional behaviour as mesenchymal cells with phagocytic activities. The variability in expression of antigens related to glial and monocytic/macrophage differentiation stressed the immunophenotypic heterogeneity of tumour cells in PXAs. PMID- 12899202 TI - The prognostic role of proliferation activity in human CNS tumours: the determination of AgNOR, PCNA and Ki-67 expression. Part 1: AgNOR expression in CNS tumours. AB - The aim of the study was an evaluation of the proliferation activity of Central Nervous System tumours by using silver colloid staining of nucleolar organiser regions (AgNORs) and comparing the results with the proliferation activity estimated by using PCNA and Ki-67 antibodies to clarify the clinicopathological significance of this method. The first part of the study describes the results of AgNOR expression and its correlation with the histological type and grading of the tumours. The study group comprised samples from 204 cases of CNS tumours and 10 cases of post-traumatic neuroma. The analysis of staining results with the use of a computerised image analysis system revealed significant differences between the expression of AgNOR in CNS tumours of different grades. A spectrum of gradually increasing AgNOR expression from benign to low and high grade tumours indicates that the AgNOR staining method can be a useful supplement to the histopathological diagnostics of CNS tumours. PMID- 12899203 TI - Neuropathological and anatomopathological analyses of acardiac and "normal" siblings in an acardiac-twin pregnancy. AB - Acardiac twinning is a very rare complication of multiple pregnancy. The authors present the neuropathological and anatomopathological description of the twins of the multiple pregnancy complicated by the acardiac foetus and terminated at 26 weeks of gestation. An anatomopathological examination of the "normal" twin showed hyaline membrane syndrome, cardiomegaly and hepatomegaly. Neuropathologically, numerous hypoxic-ischaemic lesions, most likely associated with haemodynamic disorders during pregnancy as well as less pronounced perinatal changes were revealed. The acardiac foetus, classified as acardius acephalus, demonstrated the presence of some abdominal organs and a histologically well developed spinal cord. In view of the neuropathological changes, monitoring "normal" twins for discreet pathological central nervous system signs, which may be similar in character to those described, may play a significant role. PMID- 12899204 TI - Treatment of an infantile aneurysm of the vein of Galen by total excision: a case report. AB - The paper presents a case of an aneurysm of the vein of Galen (VGA) in an infant, successfully treated by total open surgical excision (after previous ineffective embolisation). Current strategies of management of these rare yet potentially disastrous lesions are discussed and pertinent literature is reviewed. PMID- 12899205 TI - The phrenic ganglion in man. AB - During educational dissections we observed a phrenic ganglion on the nerve of the phrenic artery originating from the upper pole of the right coeliac ganglion, which accompanied the right inferior phrenic artery on a female cadaver at the age of 34. In our case the left coeliac ganglion, the inferior phrenic artery, the right and left greater, lesser and least splanchnic nerves were present and normal. However, the left nerve of the phrenic artery and the phrenic ganglion were absent. We consider that this rarely reported neural formation may be of importance for anatomists and clinicians. PMID- 12899206 TI - Stop inhaling smoke: prevent coronary heart disease. AB - Acute myocardial infarction (AMI) was rare a century ago and was diagnosed in few living patients prior to 1925. By 1950, it was the most common heart problem seen by clinicians. Thought at first to have been overlooked, there were many explanations offered for its neglect. Smoking, hypertension, and elevated cholesterol are associated with AMI, but of these only smoking should be considered a cause. Hypertension and hypercholesterolemia may be co-effects, perhaps of inflammation stimulated in the lung and blood vessels by smoking and air pollution, thus affecting vessels and arteries subjected to systemic blood pressure. Air pollution--the 20th century's other "big smoke"--deserves consideration as a 2nd cause. Auto exhaust blankets the world's cities. It consists of smoke and other effluents of petroleum vaporization and combustion that emanate from the crankcases and exhaust pipes of trucks and automobiles. The major living spaces (conurbations) of the world now imitate and exceed Los Angeles in their levels of air pollution. Auto exhaust gases fit the timeline, and their increasing amounts parallel the worldwide rise in coronary heart disease. Increasing doses of these chemicals imitate cigarette smoke and stimulate inflammation in the lungs. They appear to be absorbed into the blood, where they cause inflammation in blood vessels, increased blood pressure, and clogged coronary arteries. Avoidance is the obvious solution. Quit inhaling cigarette smoke and motor vehicle exhaust. The benefits have been shown and can be proved by intervention. The quest for clean air is hygienic-like avoiding water contaminated with feces was 150 yr ago. Clear air must be made a moral right. Its attainment requires a major revolution in priorities for energy use and lifestyle. Two types of smoke must be avoided. The world's most lethal disease. PMID- 12899207 TI - Elevated childhood cancer incidence proximate to U.S. nuclear power plants. AB - Numerous reports document elevated cancer rates among children living near nuclear facilities in various nations. Little research has examined U.S. rates near the nation's 103 operating reactors. This study determined that cancer incidence for children < 10 yr of age who live within 30 mi (48 km) of each of 14 nuclear plants in the eastern United States (49 counties with a population > 16.8 million) exceeds the national average. The excess 12.4% risk suggests that 1 in 9 cancers among children who reside near nuclear reactors is linked to radioactive emissions. If cancer incidence in 5 western states is used as a baseline, the ratio is closer to 1 in 5. Incidence is particularly elevated for leukemia. Childhood cancer mortality exceeds the national average in 7 of the 14 study areas. PMID- 12899208 TI - Water chlorination and birth defects: a systematic review and meta-analysis. AB - The authors used the Medline data retrieval system for 1966-2001 to identify and synthesize the epidemiological evidence for adverse effects of prenatal exposure to chlorination byproducts on birth defects. For 5 studies that provided estimates of the effect of exposure on 1 or several birth defects, the authors calculated summary odds ratios and 95% confidence intervals using the Mantel Haenszel method for a fixed-effects model, and the DerSimonian-Laird method for a random-effects model. The results of this meta-analysis provide evidence for an effect of exposure to chlorination byproducts on the risk of any birth defect, and of neural tube and urinary system defects. The results for respiratory system, major cardiac, and oral cleft defects were heterogeneous and inconclusive. Differences in the susceptibility of populations, and different approaches in exposure assessment, may explain the heterogeneity of the results. PMID- 12899209 TI - Molecular pathology of skin carcinogenesis due to arsenicalism from coal-burning. AB - Arsenicalism has been observed throughout the world and has become an urgent public health concern. The authors explored the mechanism of carcinogenesis of inorganic arsenic in patients with arsenicalism from coal-burning pollution. The 68 subjects were divided into 3 groups--carcinoma, precarcinoma, and common-on the basis of pathological diagnosis. The expressions of proliferating cell nuclear antigen (PCNA), mutant-type P53, and B-cell lymphoma/leukemia-2 (BCL-2) proteins were detected by immunohistochemical staining. PCNA, P53, and BCL-2 proteins were overexpressed. The proteins' overexpressions correlated with the pathological changes seen in each pathological study group (i.e., common < precarcinoma < carcinoma). Statistical correlation was observed between P53 and BCL-2, and between PCNA and BCL-2. The authors concluded that cell proliferation, antiapoptosis, and up-regulation of the mutant-type P53 gene played vital roles in the pathological development of arsenicalism. PMID- 12899210 TI - Exposure to environmental tobacco smoke in restaurants without separate ventilation systems for smoking and nonsmoking dining areas. AB - In this study, the author examined (a) levels of airborne pollutants from environmental tobacco smoke in 8 restaurants, and (b) changes in urinary cotinine and nicotine levels among 97 nonsmoking subjects (i.e., 40 restaurant employees, 37 patrons, and 20 referents). Airborne pollutant levels were significantly lower in the control environments than in the nonsmoking dining rooms in which smoking was not permitted, and the levels were significantly lower in the dining rooms in which smoking was not permitted than in the dining rooms in which smoking was permitted. Levels of urinary cotinine and nicotine increased among subjects in the dining rooms in which smoking was permitted, and the increase was significantly greater in employees than patrons. There was a significant positive correlation between levels of urinary nicotine increase and the levels of airborne nicotine and solanesol. The results of this study support the restriction of smoking to designated areas that have separate ventilation systems, or the prohibition of smoking in restaurants. PMID- 12899211 TI - Simultaneous determination of urinary cadmium, cobalt, lead, and nickel concentrations in steel production workers by differential pulse stripping voltammetry. AB - Screening for metals in urine is important because toxic levels of these elements are linked to disease. In the current study, the authors used differential pulse stripping voltammetry on a hanging mercury drop electrode for the simultaneous determination of cadmium, cobalt, lead, and nickel in the urine of 63 production and 63 quality control workers in a steel production plant, along with 63 matched normal controls. Urinary sampling is a noninvasive procedure, and in this study participants had good compliance. Metal levels for both the production and quality control workers were significantly higher than for controls. The results reveal the need for immediate improvements in workplace ventilation and industrial hygiene practices for this cohort. PMID- 12899212 TI - Health risks of residential exposure to polycyclic aromatic hydrocarbons. AB - A disease prevalence study and follow-up health surveillance were conducted among residents of an African-American community situated at the site of a former creosote wood-treatment facility contaminated with polycyclic aromatic hydrocarbons. Household interviews were conducted among 214 residents living around the hazardous waste site (target population) and 212 comparison residents in a neighborhood 2.4 km away from the site. Target area residents reported a higher prevalence of skin rashes than comparison residents (relative risk [RR] = 5.7; 95% confidence interval [CI] = 3.0, 10.9). The prevalence of reported rashes increased with increasing levels of anthracene detected in yards (test for linear trend, p = 0.02). With adjustment for environmental worry, reports of chronic bronchitis and difficulties becoming pregnant did not differ significantly between target and comparison residents (p > 0.05). PMID- 12899213 TI - Environmental factors and the proportion of males at birth in Italy. AB - A significant decline in the proportion of male births has been observed in recent decades in several western countries, and several researchers have raised the question of the possible role of environmental pollution in this trend. In the present analysis, the authors examined the effect of environmental factors (pollution from pesticides, urban pollution, and industrial pollution) on the sex ratio (proportion of males) of Italian singleton 1st births for the years 1989 1993, controlling for the age of the mother. The data show a slight increase in the sex ratio from north to south; however, no reduction in the proportion of males was seen in areas with higher pesticide consumption or intense industrial pollution. A small reduction in sex ratio was found in the major urban centers. The analysis was based on individual information on birth order and maternal age, as well as on ecological data on environmental pollution. No significant association was found between environmental pollution and the proportion of male births in Italy during the study period. PMID- 12899214 TI - Case report: further upward migration of a misplaced double-J ureteral stent. AB - A 45-year-old man was referred to our hospital for the removal of a misplaced double-J ureteral stent. The lower end of the stent shortly after insertion was at the level of sacroiliac joint. However one week later the stent was further migrated upwards to the level of 4th lumbar vertebra. Immediate ureterorenoscopy was performed to prevent further migration of the stent to the renal pelvis and the stent was removed by the grasping forceps under direct vision successfully. PMID- 12899215 TI - Endoscopic approach of the overinfected urachal cyst. AB - The overinfected urachus cyst presents itself in quite different forms. In spite of the accuracy of ultrasound and abdominal CT scan, sometimes the diagnosis of this urachal abnormality is imprecise and even misleading. This article describes the transvesical endoscopic drainage of an infected urachal cyst with optimum result upon the treatment of this condition. PMID- 12899216 TI - Polypoid cystitis unrelated to indwelling catheters: a report of eight patients. AB - Since polypoid cystitis (PC) is generally caused by indwelling catheter use, in order to evaluate the patients with PC unrelated to a intravesical catheter, a retrospective analysis of the records of the Pathology Department of Turgut Ozal Medical Center was performed and this revealed 8 patients. Mean age of the 2 female and 6 male patients was 48 years (28 to 70). None of the patients had bacterial growth in urine cultures. All cases were diagnosed incidentally by radiologic and cystoscopic examinations in the evaluation of different conditions, such as hematuria, ovarian abscess, bladder carcinoma, erectile dysfunction, neurogenic bladder, benign prostate hyperplasia and unexplained dysuria. At the beginning, all patients were diagnosed mistakenly as bladder carcinoma. The definitive diagnosis was made after histopathologic examinations of transurethrally resected specimens. Patients were followed for 6 months to 2 years after first diagnosis. No recurrence was established during follow-up. The final urologic examinations which were done currently, were normal. In conclusion, PC is a benign lesion and should be considered in the differential diagnosis of transitional cell carcinoma of the bladder. PMID- 12899217 TI - Acute pyelonephritis causing acute renal allograft dysfunction. AB - In renal transplant recipients, acute pyelonephritis may cause acute deterioration of renal function. We report a case with acute allograft failure due to acute pyelonephritis, which was confirmed by graft biopsy. After appropriate antimicrobial therapy, allograft function recovered. PMID- 12899218 TI - Fluid distribution measurement between body compartments in primates in disclosing fluid retention during prolonged hypokinesia. AB - Body fluid homeostasis undergoes significant changes during hypokinesia (diminished movement). Understanding of fluid transfer between body fluid compartments and its regulating mechanisms was the aim of this study for disclosing impaired fluid retention during hypokinesia (HK). Studies were done on 12 male Macaca mulatta (rhesus monkeys) aged three to five years (5.15 to 6.56 kg) during 90 days period of pre-HK and 90 days period of HK. All primates were divided equally into two groups: vivarium control primates (VCP) and hypokinetic primates (HKP). Hypokinetic primates were kept for 90 days in small individual cages that restricted their movements in all directions without hindering food and fluid intakes. Control primates were housed in individual cages without their movements being restricted. Total body fluid (TBF), intracellular fluid volume (IFV) and circulating plasma volume (CPV) decreased significantly (p < 0.05), while extracellular fluid volume (EFV) and interstitial fluid volume (IsFV) decreased significantly (p < 0.05) at the initial seven days of the HK period and after the 7th day increase progressively in HKP compared with their pre-HK values and their respective control values in VCP. Fluid excretion, urinary and plasma sodium (Na) and potassium (K) levels increased significantly (p < 0.05), while fluid intake and fluid retention decreased significantly (p < 0.05) in HKP compared with their pre-HK values and their respective controls in VCP. The measured parameters did not change in VCP compared with their pre-HK values. It was concluded that decreased IFV and CPV may demonstrate fluid depletion, while marked increase of fluid loss despite of fluid depletion may demonstrate impair fluid retention during HK. Dissociation between fluid loss and fluid depletion may demonstrate the presence of reduced fluid retention as the mechanism of development of fluid depletion during HK. PMID- 12899219 TI - Psoas abscess twenty-one years after ipsilateral nephrectomy. AB - We report an unusual case of psoas abscess, which developed twenty-one years after ipsilateral nephrectomy and was caused by infrequent pathogen, Proteus mirabilis. It was diagnosed by computed tomography and was drained percutaneously with a nephrostomy tube guided by ultrasonography. PMID- 12899220 TI - Vascular plexus is a differentation criterion for muscularis mucosa from muscularis propria in small biopsies and transurethral resection materials of urinary bladder? AB - OBJECTIVE: Smooth muscle fibres are found within the superficial lamina propria of urinary bladder (Muscularis Mucosa). These muscle fibres of muscularis mucosa should be distinguished from the muscularis propria in cases with urothelial carcinoma, because the depth of infiltration affects prognosis and therapy modalities. The aim of the present study was to evaluate whether the presence of vascular plexus is a criterion for muscularis mucosa and to distinguish it from muscularis propria in urinary bladder biopsies and transurethral resection specimens. MATERIALS AND METHODS: Hematoxylin-Eosin and Masson-Trichrome stained slides of 54 cases with urothelial carcinoma were reviewed. RESULTS: In all cases (100%), thick walled vessels were observed within the lamina propria of urinary bladder. Smooth muscle layer of lamina propria was seen in 48 of 54 cases (88.8%), and these smooth muscle fibres were co-existed with thick walled vessels. In invasive urothelial carcinoma; thin smooth muscle fibres and thick walled vessels were seen within the lamina propria in 22 of 24 cases (91.7%). There were two cases (8.3%) including only vessels in small biopsies of invasive urothelial carcinoma cases. In cases with non-invasive urothelial carcinoma; muscle fibres of muscularis mucosa and vessels were found within the lamina propia in 26 of 30 cases (86.7%). In this group, muscle fibres were not detected in 4 cases (13.3%). CONCLUSION: Muscularis mucosa can be detected histologically in urinary bladder biopsies and TUR materials. We emphasize that the presence of vascular network is a useful criterion to determine muscularis mucosa of urinary bladder, even in the absence of muscle fibres. PMID- 12899221 TI - The effects of piroxicam to the muscosal barrier of the bladder after overdistension--an experimental study in rabbits. AB - The effects of piroxicam on the duration and severity of damage to the mucosal barrier of the urinary bladder after overdistension was investigated. Overdistension of the bladder was induced for 3 h in 16 New Zealand male rabbits by giving Ringer's lactate infusion (40 mL/kg/h) and furosemide (1 mg/kg) to the peritoneal cavity. Insertion of 8 Fr Foley catheter was used for obstruction of the bladder neck. In both control (C) and piroxicam (P) groups, 20 mL of 2% solution of Trypan blue in 0.9% NaCl solution was instilled into the bladder for 1 hr at 0, 24, 48 h and 7 days after overdistension. In group P daily intramuscular injection of 5 mg/kg piroxicam, and isotonic saline in group C was administrated for 7 days. Full-thickness samples were taken from the bladder at 0, 24, 48 h and 7 days after overdistension. The bladder wall was deep blue throughout in both groups at 0, 24, and 48 h. The severity and duration of inflammatory reaction was lower and nearly normalized on the 7th post procedure day in piroxicam group. CONCLUSION: Inflammatory reaction can be prevented by administration of anti-inflammatory drugs such as piroxicam but the prevention of increased permeability is unclear after overdistension of the bladder. PMID- 12899222 TI - Autocystectomy following extensive genitourinary tuberculosis: presentation and management. AB - Genitourinary tuberculosis is an important cause of morbidity in developing and developed countries. We describe a case of extensive genitourinary tuberculosis in which there was complete destruction of the right kidney, stricture of the entire left ureter and an autocystectomized bladder. He presented in azotemia with urinary incontinence and was managed by right nephroureterectomy, ileal replacement of left ureter and ileal neobladder after a preliminary nephrostomy and antituberculous chemotherapy. PMID- 12899223 TI - Evaluation of multiple recurrence events in superficial bladder cancer patients treated with intravesical bacillus Calmette-Guerin therapy using the Andersen Gill's model. AB - To evaluate factors affecting recurrence after intravesical bacillus Calmette Guerin (BCG) therapy (Tokyo 172 strain), we reviewed data for 101 patients with superficial bladder cancer (pTa [n = 80] and pT1 [n = 21]) treated between 1985 and 1999. The median follow-up period was 58.9 months. Factors affecting the first tumour recurrence were evaluated using Cox's proportional hazards model and those affecting multiple recurrence with Andersen-Gill's model. The 5-year recurrence-free rate was 63% for all 101 patients. The recurrence frequency, defined as times per 100 patient-months of follow-up, greatly decreased from 7.3 +/- 9.6 (SD) before the instillation to 2.6 +/- 5.6 after the therapy (p < 0.0001). Patients with pT1 tumours tended to have earlier recurrence than those with pTa tumours (p = 0.06). Multivariate analysis using Cox's proportional hazards model revealed that a history of bladder cancer and pathological stage were independent factors affecting the first tumour recurrence after the BCG therapy. When multiple endpoints of recurrence were evaluated using the Andersen Gill's model, number of tumours as well as a history of bladder cancer and pathological stage demonstrated significant links to tumour recurrence after the BCG therapy. The 5-year progression-free and 5-year survival rates were 89.3% and 85.3% for all the 101 patients, respectively. Because intravesical recurrence may involve multiple events during the clinical course of patients with bladder cancer, the Andersen-Gill's model appears useful for evaluation of risk factors. PMID- 12899225 TI - Prostatic atrophy: evidence for a possible role of local ischemia in its pathogenesis. AB - Prostatic atrophy may be histologically and at ultrasound similar to adenocarcinoma causing diagnostic confusion, its frequency increases with age but the etiopathogenesis is unknown. Based on a systematic study in autopsies previously done by one of us, ischemia due to local intense arteriosclerosis seems to be a potential factor for its pathogenesis. Absent blood flow in areas of prostatic atrophy might be a further evidence for a possible role of ischemia. From a total of 298 patients biopsied and studied by gray-scale and color Doppler transrectal ultrasound in the period 1998 to 2001, 33 patients had suspicious lesions (37 hypoechoic nodules and 3 heterogeneous lesions) showing prostatic atrophy as the only diagnosis on all these biopsied lesions. Adenocarcinoma, high grade intraepithelial neoplasia or other atypical lesions were absent in all patients. On color Doppler the suspicious areas showed absent flow in 24/40 (60%), present flow in 12/40 (30%), and increased flow in 4/40 (10%) of the lesions. Absent flow in the majority of the lesions studied may be a further evidence for a possible role of local ischemia in the etiopathogenesis of prostatic atrophy. PMID- 12899224 TI - Vesicouterine fistulas following cesarean section: report on a case, review and update of the literature. AB - Herein we report on 1 more case of vesicouterine fistula following cesarean section with review and update of the literature concerning this unusual topic. The disease presented with vaginal urinary leakage, cyclic hematuria and amenorrhea. The fistula was successfully repaired by delayed surgery. Actually, all over the world the prevalence of the disease is increasing for the frequent use of the cesarean section. Fistulas may develop immediately after a cesarean section, manifest in the late puerperium or occur after repeated procedures. Spontaneous healing is reported in 5% of cases. Vesicouterine fistulas present with vaginal urinary leakage, cyclic hematuira (menouria), amenorrhea, infertility, and first trimester abortions. The diagnosis is ruled out by showing the fistulous track between bladder and uterus as well as by excluding other more frequent urogenital fistulas. The disease treatment options include conservative treatment as well as surgical repair. Rarely, patients refuse any kind of treatment because of the benignity of symptoms and prognosis of the disease. Conservative management by bladder catheterization for at least 4-8 weeks is indicated when the fistula is discoveredjust after delivery since there is good chance for spontaneous closure of the fistulous track. Hormonal management should be tried in women presenting with Youssef's syndrome. Surgery is the maninstay and definitive treatment of vesicouterine fistulas after cesarean section. Patients scheduled for surgery should undergo pretreatment of urinary tract infections. Surgical repair of vesico-uterine fistulas are performed by different approaches which include the vaginal, transvesical-retroperitoneal and transperitoneal access which is considered the most effective with the lowest relapse rate. Recently, laparoscopy has been proposed as a valid option for repairing vesicouterine fistulas. The endoscopic treatment may be effective in treating small vesicouterine fistulas. The pregnancy rate after repair is 31.25% with a rate of term deliveries of 25%. The disease may be prevented by emptying the bladder as well as by carefully dissecting the lower uterine segment. It is advisable that after vesicouterine fistula repair delivery should be performed by repeating a cesarean section since the risk of fistula recurrence. Usually, vesicouterine fistulas are diagnosed postoperatively. As a result, at least 95% of patients will undergo another operation for repairing the fistula. In the meantime they are bothered by related symptoms which impair their quality of life. As far as we are concerned intraoperative diagnosis is the gold standard in detecting vesicouterine fistulas for allowing immediate repair. We propose intraoperative sonography by the transvaginal (or transrectal) route for the Foley transurethral catheter producing bloody urine, for suspecting bladder injury while dissecting the uterine lower segment and for monitoring patients who already had had vesicouterine fistula repair. As a result patients will avoid the familial and social problems related to the disease as well another operation. Moreover, ultrasound Doppler examination may help in better investigating and understanding the pathophysiology of vesicouterine fistulas. PMID- 12899226 TI - Lower urinary tract symptoms, pain and quality of life assessment in chronic non bacterial prostatitis patients treated with alpha-blocking agent doxazosin; versus placebo. AB - The efficacy of doxazosin monotherapy in chronic non-bacterial prostatitis was investigated in terms of urinary symptom, pain and quality of life assessment versus placebo. A total of 60 men with chronic non-bacterial prostatitis were randomised to daily supplement of 4 mg doxazosin or a placebo, for 3 months. International Prostate Symptom Score (IPSS) questionnaire was self administered at the entry and at 3 months after the cessation of the treatment. In addition, patients were asked to complete 2-item questionnaire on pain related symptoms of chronic prostatitis. Quality of life was assessed with a single item included in IPSS. Three months after cessation of the treatment there was a significant difference between the overall mean IPSS, pain and quality of life scores of the two groups in favour of alpha-blocking agent use (p = 0.001, p < 0.001 and p < 0.001, respectively). In patients undergone doxazosin treatment; symptom, pain and quality of life status revealed 32.94 +/- 5.27%, 36.57 +/- 5.67% and 36.78 +/ 4.75% overall improvement, respectively. IPSS appeared to be a valuable tool in assessing treatment outcome of chronic non-bacterial prostatitis. PMID- 12899227 TI - Incidence of neuroendocrine cells in the seminal vesicles and the prostate--an immunohistochemical study. AB - INTRODUCTION: Prostate and seminal vesicles (sv) are both androgen-dependent male sex accessory glands. The cancer incidence in these two organs is vastly different. Neuroendocrine (NE) cells are involved in the regulation of prostate growth, differentiation and in prostate cancer carcinogenesis. Thus, knowledge of the incidence of NE cells in sv may add to our understanding of prostate cancer etiology. METHODS: Samples of histologically confirmed normal prostate tissue and normal sv tissue from 20 men were immunostained for chromogranin A. The incidence of stained cells was evaluated semiquantitately. RESULTS: Neuroendocrine cells were detected in all prostate specimens, but not a single stained cell was found in any of the sv specimens. CONCLUSIONS: The lack of NE cells and, subsequently, of biogenic amines, peptides and growth factors may be a reason for the low cancer incidence in the seminal vesicles. Alternatively, the absence of NE cells can be seen as a hint that the stem cells of the prostate and sv react differently to endogenous and exogenous stimuli, and thus in the seminal vesicles, stem cells are not transformed into NE cells. PMID- 12899228 TI - Synchronous diagnosis of prostatic adenocarcinoma and sarcoidosis. AB - Sarcoidosis and prostatic carcinoma in the same patient is a rare clinical entity. This is the third reported case of such pathology. Immunodeficiency caused by sarcoidosis may induce immunity disorientation resulting in the presence of prostatic carcinoma. Sarcoidosis in relation with malignancy may give some answers for the etiopathology of cancer. PMID- 12899229 TI - Ascites following prostatectomy: a rare presentation of comorbid disease. AB - We report a rare case of ascites developing in the early post-prostatectomy period due to a mass originating from the head of the pancreas. Comorbid diseases that are not documented preoperatively may emerge with signs mimicking the complication of prostate surgery. PMID- 12899230 TI - Lycopene therapy in idiopathic male infertility--a preliminary report. AB - Excessive generation of reactive oxygen species (ROS) containing free oxygen radicals has been identified as one of the causes of male infertility. Lycopene is a component of human redox defence mechanism against free radicals. It is found in high concentrations in the testes and seminal plasma and decreased levels have been demonstrated in men suffering from infertility. We evaluated the effect of oral lycopene therapy in men with idiopathic infertility. Beginning March 2000, thirty men with idiopathic non-obstructive oligo/astheno/teratozoospermia were enrolled for the trial. All patients were administered 2000 mcg of Lycopene, twice a day for three months. Semen analysis was performed at three months and sperm concentration, motility and morphology were evaluated. All patients completed the trial without any complications. Twenty patients (66%) showed an improvement in sperm concentration, sixteen (53%) had improved motility and fourteen (46%) showed improvement in sperm morphology. In cases showing an improvement, the median change in concentration was 22 million/ml, motility 25% and morphology 10%. The improvement in concentration and motility were statistically significant. Baseline sperm concentration less than 5 million/ml was associated with no significant improvement. Higher baseline concentrations were associated with significant improvement and resulted in six pregnancies in 26 patients (23%). Oral Lycopene therapy seems to have a role in the management of idiopathic male infertility. Maximum improvement seems to occur in the sperm concentration (66% cases). Patients without severe oligospermia (sperm density > 5 million/ml) may be given a trial of therapy with lycopene. However, larger randomised controlled trials are essential before definitive therapeutic guidelines can be made. PMID- 12899231 TI - Solitary neurofibroma of the spermatic cord. AB - We report a case of solitary neurofibroma of the spermatic cord. Such benign tumors arise from perineural and Schwann cells and may be located in various parts of the body, but are rarely observed in the spermatic cord. No clinical or laboratorial manifestations of von Recklinghausen disease (Neurofibromatosis) were identified. Distinct criteria have been established for a diagnosis of von Recklinghausen disease, so that a solitary neurofibroma may not represent this complex. Only a little number of solitary neurofibromas of the spermatic cord are reported in the literature. PMID- 12899232 TI - Urethral fistula caused by acne inversa (hidradenitis suppurativa): a case report. PMID- 12899233 TI - The utility of lockout valve reservoirs in preventing autoinflation in penile prostheses. AB - INTRODUCTION: Autoinflation is a troublesome complication following penile prosthesis placement that may be potentiated by prevesical scarring following radical prostatectomy. We evaluated the frequency of autoinflation and other complications following penile prosthesis placement in radical prostatectomy patients and controls as a surrogate to establishing the utility of lockout reservoirs in preventing autoinflation. METHODS: 139 prostheses (including 14 with lockout reservoirs) were placed in 132 men (including 35 post-prostatectomy patients) over a 5(1/2) year period at our institution. Outcomes assessed include postoperative complications and the need for revision or replacement of the prosthesis. Multivariable regression analysis was used to determine the association of patient, device-specific, and perioperative characteristics with these outcomes. RESULTS: There was no difference in the postoperative complication and re-operation rates between post-prostatectomy patients and controls (both p > 0.77). The incidence of autoinflation in post-prostatectomy patients and controls was 3% and 5%, respectively (p > 0.99). Patients with prior prostheses were 3 times as likely to develop a postoperative complication or require prosthesis revision (p = 0.02). CONCLUSION: Penile prostheses are well tolerated in post-prostatectomy patients with comparable outcomes to those men with organic erectile dysfunction. The frequency of autoinflation does not appear to be increased in post-prostatectomy patients. Initial results with the lockout valve reservoir in preventing autoinflation are encouraging though additional study is warranted to justify their routine use. PMID- 12899234 TI - Acute scrotum secondary to filarial infection: a case report. AB - A 23-year-old man immigrated from Sri Lanka came to our observation for an acute painful volume increase of the right scrotum without fever. Clinical examination suggested a diagnosis of testis torsion. An exploratory surgical procedure was performed. An inflammatory spermatic cord and epididymis with a purple nodule of the middle portion were found. The nodule was excised and sent to pathologist that diagnosed a filarial infection. The patient was successfully treated with diethylcarbamazine. PMID- 12899235 TI - A life-threatening infection: Fournier's gangrene. AB - Fournier's gangrene is a life-threatening disorder in which infection of the perineum and scrotum spreads along fascial planes, causing soft tissue necrosis. If urgent surgery is delayed, the disease will soon result in septic shock, multiorgan failure, and death. In this study, we present 21 patients with Fournier's gangrene who were treated in period between 1994 and 2001. Patients' charts were reviewed retrospectively and are discussed in the light of literature. All patients received aggressive surgical debridment. Penicillin or Ceftriaxone, aminoglicoside and metronidazole were administered intravenously. Of the 21 patients, 5 had scrotal carbuncle, 1 had urethral stricture, 1 had chronic indwelling urethral catheterization, 2 had perirectal abscess, and 1 had hemorrhoidectomy. In eleven patients we couldn't identify any cause. Twelve patients had diabetes mellitus, and two had chronic alcoholism. Escherichia coli was isolated in 12 purulent tissue cultures, and Bacteroides fragilis in eight. Seventeen patients survived, whereas four died. Fourier's gangrene is considered a surgical emergency. Early surgical intervention is essential, as the gangrene can spread rapidly at rates reaching 2 mm per hour. So that Fournier's gangrene is an abrupt, rapidly progressive, gangrenous infection of the external genitalia and perineum and is a real urologic emergency. PMID- 12899236 TI - Beyond Kt/V: redefining adequacy of dialysis in the 21st century. AB - I presume that every dialysis patient needs a minimal amount of dialysis, since less dialysis will lead to death. Until we come up with a better index, for hemodialysis, I propose that we should return to Scribner's 1974 recommendation on adequacy of dialysis and concentrate on the homeostatic function of the kidney, as indicated in Table 1. For those who still want to measure dialysis dose by Kt/V, the optimal dose for HD is a Kt/V of 1.2/treatment and for PD 1.7/week. None of these would be adequate without good UF and control of patients' fluid status. Frequent (4, 5, 6 or 7 days/week) and prolonged dialyses (5-8 hours) should be the norm and I believe that the Hemodialysis Product should be used as a guide, pending confirmation by prospective studies. A high hemodialysis product reflects much more closely the function of the normal kidneys than 3-4 hours of dialysis three times a week with a Kt/V of 1.2 per session. Peritoneal dialysis that is already continuous needs to improve its efficiency. For PD patients, I believe that the total daily volume (Kt) might be a better index of adequacy, especially if it is combined with a good ultrafiltration (at least of 1 L/day) and blood pressure control. Of course, this has to be confirmed in the future. In the anuric patients, the daily volume can be kept to 8 L/day if the patient is free of symptoms, but should be increased to 10-12 L/day for CAPD and 15-20 L/day for APD patients if they develop symptoms of underdialysis and fluid overload. PMID- 12899237 TI - Peritoneal dialysis in the nursing home. AB - BACKGROUND: During the past few decades, the demographics of end stage renal disease have been changed significantly with the emerging predominance of elderly patients. Elderly dialysis patients are usually more dependent and may need long term placement in a long-term care facility. Failure to meet the needs of these patients may have a significant impact on the peritoneal dialysis program. We report our experience of starting peritoneal dialysis program in a community based Long Term Care Facility (LTCF). METHODS: During the period of 2000-2001, after appropriate training of nursing home personnel, we admitted 8 peritoneal dialysis patients to one community-based nursing home. All information presented here has been collected through chart review. RESULTS: At the time of admission the average age of the 8 patients was 77.3 +/- 7.2 years (range 69 to 91 years). All patients had several comorbid diseases and six of the eight were bed-ridden. The patients stayed in the facility for a total of 29.57 patient months. One patient had three episodes of peritonitis within three months (all culture negative) and has been excluded from the analysis of the overall peritonitis rate. The peritonitis rate for the other seven patients was 1 per 7.54 patient month. Six patients were readmitted to hospital because of peritonitis, severe malnutrition and hip fracture. Four of them died in the hospital. One died in the nursing home. One patient remains in the nursing home at the present time. CONCLUSIONS: Our experience suggests that peritoneal dialysis can be achieved in a community-based nursing home. This requires a systematic training program for the LTCF personnel and the availability of a "dedicated" nephrology dialysis staff. This is crucial to the success of the program. It is important that patients, their families and ESRD care professionals are informed of the limited survival expectation particularly for very old and severely impaired patients. PMID- 12899238 TI - Failure of intravenous fluid therapies to decrease serum sodium levels in elderly hospitalized patients. AB - Elderly patients may have a tendency to develop hyponatremia due to sensitivity to stimuli that release ADH as well as an impaired ability to excrete a water load. We evaluated changes in serum sodium in elderly hospitalized patients who received various forms of intravenous fluid therapies. All patients were required to have a baseline serum sodium of 136-145 meq/L. Fourteen patients were enrolled in the study. The mean age was 82.9 +/- 6.8 years (mean +/- SEM). Thirty-six % were nursing home residents. Seventy-nine % were females. Seventy-two % received half normal saline and the remainder received normal saline as intravenous fluid therapy. The patients received a mean of 1098 +/- 145 mL of intravenous fluid per day, in addition to oral fluids. Mean follow up period was 5.9 days (3-10 days). Mean baseline serum sodium was 140.2 +/- 0.7 meq/L andmean follow up serum sodium was 141.4 +/- 0.9 meq/L. The m ean baseline BUN was 25 +/- 3.6 mg/dL and mean follow u BUN was 19.6 +/- 3.4 mg/dL. The mean baseline serum creatinine was 0.9 +/- 0.1 mg/dL and mean follow up creatinine was 0.9 +/- 0.1 mg/dL. The postintravenous fluid therapy serum sodium in the group receiving half normal saline was 141.7 +/- 0.7 meq/L and 140.8 +/- 3 meq/L in the normal saline group. No significant difference was observed between the pre and post fluid therapy for any of these paramenters (p > 0.05). Mean baseline plasma renin activity was 1.6 +/- 0.7 ng/ml/hour and fifty-seven % had PRA of less than 1 ng/ml/hour. Mean plasma aldosterone was 8.5 +/- 1.8 ng/mL and forty-two % were less than 5.5 ng/mL. Plasma ADH and ANP was 5.7 +/- 3.4 pg/mL and 83.6 +/- 26.9 pg/mL, respectively. Mean serum and urine osmolalities were 290 +/- 3.1 mOsm/kg and 471 +/- 57.7 mOsm/kg, respectively. No patient developed hyponatremia and 7 of the 14 patients experienced an increase in serum sodium during the follow up period. We conclude that many elderly patients hospitalized for acute medical illnesses either maintain a stable serum sodium or experience an increase in serum sodium. This occurs because total fluids administered to these patients are generally insufficient. PMID- 12899239 TI - Pyocystis in patients on chronic dialysis. A potentially misdiagnosed syndrome. AB - Pyocystis is an important complication of non-functioning urinary bladder, which often poses diagnostic difficulties. We present a case of pyocystis in a patient on chronic hemodialysis who was anuric for one year. The patient was initially diagnosed with diverticulitis. An abdominal C-T scan suggested the diagnosis of pyocystis, which was confirmed by bladder catheterization. The patient was treated with bladder drainage and a prolonged antibiotic course, followed by intermittent saline washing of the bladder. PMID- 12899240 TI - Clinical pharmacology physiology conference: metformin and lactic acidosis (LA). PMID- 12899241 TI - Understanding the selectivity of fumagillin for the methionine aminopeptidase type II. AB - The aim of this study is to explain the selectivity of the antiangiogenic drug fumagillin for the eukaryotic enzyme methionine aminopeptidase type II (MetAP-II, EC 3.4.11.18) over the structurally very similar MetAP-I. A homology model for the human MetAP-I is constructed and molecular dynamics simulations are performed on this model with and without a docked fumagillin molecule. These simulations are compared with analogous simulations that were performed on the experimentally determined structure of the human MetAP-II enzyme. We observe an increased flexibility of the active site histidine that is covalently modified by fumagillin in the MetAP-I enzyme. The MetAP-I active site residues, particularly the fumagillin-binding histidine, have a lower probability to be in a conformation that is prone to react with the drug than their MetAP-II counterparts. This result offers an explanation for the selectivity of fumagillin for the eukaryotic MetAP-II enzyme. PMID- 12899242 TI - In vitro antitumor SAR of threo/cis/threo/cis/erythro bis-THF acetogenins: correlations with their inhibition of mitochondrial Complex I. AB - Annonaceous acetogenins (ACG) are a large family of natural products that have been described as the most potent in vitro inhibitors of the mitochondrial respiratory chain Complex I. During the last two decades a large number of related structures have been discovered, increasing the number of members of this family. The large diversity of structural moieties and the general trends observed for inhibiting both growth of tumor cell lines and mitochondrial respiratory chain activity have resulted in the classification of these compounds into several structural groups according to their potency. Among them, the adjacent bis-tetrahydrofuranic acetogenins (bis-THF ACG) with a threo/cis/threo/cis/erythro relative configuration, have been described as the most potent subgroup, the prototypical member of which, rolliniastatin-1, was originally isolated from Rollinia membaranacea seeds. In this report we describe the different structure-activity relationships (SAR) observed for some natural ACG and semisynthetic derivatives as growth inhibitors of human tumor breast, lung, liver, and colon cell lines. All the compounds assayed showed potencies in the micromolar range. Trends observed in the cytotoxicity assay have been compared with previous data reported for these compounds as inhibitors of mitochondrial respiratory chain. PMID- 12899243 TI - SAR: flavonoids and COX-2 inhibition. AB - An analysis based upon structure-activity relationships (SAR) of the COX-2 inhibiting properties of flavonoids, a group of potential cancer chemopreventative agents, reveals that there is a dual structural basis for these activities. Each of these structural determinants (pharmacophores) alone is sufficient for activity. One of the pharmacophores is a 2D 6.9 A distance descriptor that spans the A and C rings and includes the 4-OXO and 7-hydroxyl moieties. The potency associated with that pharmacophore is determined by a series of structural modulators that can increase, decrease, or even abolish the COX-2-inhibiting potential associated with that pharmacophore. The second pharmacophore describes a para-substituted phenolic B ring that requires unsubstituted meta and ortho positions. Based upon this, it indicates that hydroxylation at the 4'-position and a free 5'-position are sufficient for COX-2 inhibiting activity. The potency associated with this pharmacophore is modulated by log P2 and by the molecular weight. PMID- 12899244 TI - Synthesis and antiproliferative activity of basic ethers of 1,2 dihydropyrrolo[1,2-a]indole, 6H-isoindolo[2,1-a]indole, and 6H benz[5,6]isoindolo[2,1-a]indole. AB - Monobasic ethers of 1,2-dihydropyrrolo[1,2-a]indole, 6H-isoindolo[2,1-a]indole, and 6H-benz[5,6]isoindolo[2,1-a]indole and bis-basic ethers of 6H-isoindolo[2,1 a]indole were prepared using an intramolecular Wittig cyclization as a key step. All these compounds were firstly evaluated for their cytotoxicity effects against L1210 cell line. Only the tetracyclic bis-basic ether 14d displayed submicromolar cytotoxic effect. Moreover, despite the fact that the presence of these two amino side chains in 14c, 14d, and 14f led to strong DNA binding effect, they are not topoisomerase II inhibitors. Among the monobasic ethers 14a, 14b, 22, and 29, which do not bind to DNA, the pentacyclic analog 29 exhibited micromolar cytotoxic activity against L1210 and HT-29 cell lines and induced a weak topoisomerase II inhibition. PMID- 12899245 TI - Molecular and functional characterization of the extracellular calcium-sensing receptor in human colon cancer cells. AB - Presence of a functional extracellular calcium-sensing receptor (CaR) is of particular relevance for the growth-inhibitory action of Ca2+ on human colon carcinoma cells. In order to detect CaR gene alterations that may have occurred during the tumorigenic process, we applied Southern blot, DNA sequence, and RT PCR analysis to DNA from normal human colon mucosa and from cancerous lesions of different grading, as well as from primary cultured and established colonic carcinoma cell lines (e.g., Caco-2). No evidence was obtained for mutations or other sequence alterations in the CaR gene in any of the colon carcinoma cells analyzed. Only a differential expression of two splice variants of the CaR gene, which are generated by usage of different promoters in the 5'-untranslated region, was detected in colon carcinomas of different grade. From Western blot analysis a tendency towards lower CaR protein levels in carcinoma cells in parallel with tumor progression became apparent. Activation of the CaR by extracellular Ca2+ or by specific receptor agonists resulted in substantial growth inhibition in Caco-2 cells. Activation of the CaR was transduced into inhibition of phospholipase A2-mediated arachidonic acid formation, but also into increased production of cAMP and IP3. This provides evidence for a cell type specific function of the CaR in human colonocytes. We conclude that neoplastic colon epithelial cells can respond to antimitogenic signals generated by activation of the CaR as long as they express sufficient amounts of the CaR protein. This provides a rationale for the use of calcium in chemoprevention of colon tumor development. PMID- 12899246 TI - Synergistic interaction between cyclophosphamide or paclitaxel and the bioreductive compound NLCPQ-1, in vivo. AB - The antitumor effect of cyclophosphamide (CPM) and paclitaxel was investigated in BALB/c mice bearing EMT6 tumors, in combination with the bioreductive compound NLCPQ-1 by using the in vivo/in vitro assay as the endpoint. An optimum administration schedule for a synergistic interaction between NLCPQ-1 and CPM/paclitaxel was determined and dose modification factors (DMF) were calculated for antitumor effect and bone marrow toxicity. All drugs were given by IP injection; NLCPQ-1 at 15 mg/kg, which is much less than its maximally tolerated dose (MTD greater than 50 mg/kg), paclitaxel up to 25 mg/kg, and CPM up to 200 mg/kg. Bone marrow toxicity studies were performed in parallel by using a modified CFU-GM assay. A schedule-dependent synergistic interaction was observed for both chemotherapeutic agents combined with NLCPQ-1 but with entirely different patterns, as has been previously seen with the analog NLCQ-1. The optimal degree of potentiation, P (percentage of tumor cells that were killed due to clear potentiation), was 31 and 33 when NLCPQ-1 was administered 2 h before CPM and 3-3.5 h after paclitaxel, respectively. At the above time schedules, NLCPQ-1 modified the dose of CPM and paclitaxel, for 60% tumor cell killing, by a factor of 1.8 and 2.1, respectively. Bone marrow toxicity was not enhanced by combining either chemotherapeutic agent with NLCPQ-1. Comparison with results from previous similar studies with NLCQ-1 revealed that, on a molar basis, NLCPQ 1 was a less potent chemosensitizer than NLCQ-1. However, the results still suggest a potential clinical use of NLCPQ-1 as an adjuvant to CPM or paclitaxel therapy against solid tumors. PMID- 12899247 TI - Brain-computer interface technology: a review of the Second International Meeting. AB - This paper summarizes the Brain-Computer Interfaces for Communication and Control, The Second International Meeting, held in Rensselaerville, NY, in June 2002. Sponsored by the National Institutes of Health and organized by the Wadsworth Center of the New York State Department of Health, the meeting addressed current work and future plans in brain-computer interface (BCI) research. Ninety-two researchers representing 38 different research groups from the United States, Canada, Europe, and China participated. The BCIs discussed at the meeting use electroencephalographic activity recorded from the scalp or single-neuron activity recorded within cortex to control cursor movement, select letters or icons, or operate neuroprostheses. The central element in each BCI is a translation algorithm that converts electrophysiological input from the user into output that controls external devices. BCI operation depends on effective interaction between two adaptive controllers, the user who encodes his or her commands in the electrophysiological input provided to the BCI, and the BCI that recognizes the commands contained in the input and expresses them in device control. Current BCIs have maximum information transfer rates of up to 25 b/min. Achievement of greater speed and accuracy requires improvements in signal acquisition and processing, in translation algorithms, and in user training. These improvements depend on interdisciplinary cooperation among neuroscientists, engineers, computer programmers, psychologists, and rehabilitation specialists, and on adoption and widespread application of objective criteria for evaluating alternative methods. The practical use of BCI technology will be determined by the development of appropriate applications and identification of appropriate user groups, and will require careful attention to the needs and desires of individual users. PMID- 12899248 TI - ERPs evoked by different matrix sizes: implications for a brain computer interface (BCI) system. AB - A brain-computer interface (BCI) system may allow a user to communicate by selecting one of many options. These options may be presented in a matrix. Larger matrices allow a larger vocabulary, but require more time for each selection. In this study, subjects were asked to perform a target detection task using matrices appropriate for a BCI. The study sought to explore the relationship between matrix size and EEG measures, target detection accuracy, and user preferences. Results indicated that larger matrices evoked a larger P300 amplitude, and that matrix size did not significantly affect performance or preferences. PMID- 12899249 TI - Use of the evoked potential P3 component for control in a virtual apartment. AB - Virtual reality (VR) may prove useful for training individuals to use a brain computer interface (BCI). It could provide complex and controllable experimental environments during BCI research and development as well as increase user motivation. In the study reported here, we examined the robustness of the evoked potential P3 component in virtual and nonvirtual environments. We asked subjects to control several objects or commands in a virtual apartment. Our results indicate that there are no significant differences in the P3 signal between subjects performing a task while immersed in VR versus subjects looking at a computer monitor. This indicates the robustness of the P3 signal over different environments. For an online control task, the performance in a VR environment was not significantly different from performance when looking at a computer monitor. There was, however, a more significant result when the subject's head view of the virtual world was fixed (p < 0.05) when compared with looking at a computer monitor. We also found that subjects' self-reported qualitative experiences did not necessarily match their objective performance. Six out of nine subjects liked the VR environment better, but only one of these subjects performed the best in this environment. The possible ramifications of this, as well as plans for future work, are discussed. PMID- 12899250 TI - Developing wearable bio-feedback systems: a general-purpose platform. AB - Microprocessors, even those in PocketPCs, have adequate power for many real-time biofeedback applications for disabled people. This power allows design of portable or wearable devices that are smaller and lighter, and that have longer battery life compared to notebook-based systems. In this paper, we discuss a general-purpose hardware/software solution based on industrial or consumer devices and a C++ framework. Its flexibility and modularity make it adaptable to a wide range of situations. Moreover, its design minimizes system requirements and programming effort, thus allowing efficient systems to be built quickly and easily. Our design has been used to build two brain computer interface systems that were easily ported from the Win32 platform. PMID- 12899251 TI - The thought-translation device (TTD): neurobehavioral mechanisms and clinical outcome. AB - The thought-translation device (TTD) consists of a training device and spelling program for the completely paralyzed using slow-cortical brain potentials (SCP). During the training phase, the self-regulation of SCPs is learned through visual auditory feedback and positive reinforcement of SCPs; during the spelling phase, patients select letters or words with their SCPs. A psychophysiological system for detection of cognitive functioning in completely paralyzed patients is an integral part of the TTD. The neurophysiological and anatomical basis of SCP regulation was investigated by recording of BOLD-response in functional magnetic resonance imaging. Results showed involvement of basal ganglia and premotor cortex for required SCP positivity. The clinical outcome of 11 paralyzed patients using the TTD and quality of life of severely paralyzed patients is described. First attempts to improve learning of brain regulation with transcranial magnetic stimulation were successful. PMID- 12899252 TI - Current trends in brain-computer interface research at the Neil Squire Foundation. AB - The Neil Squire Foundation (NSF) is a Canadian nonprofit organization whose purpose is to create opportunities for independence for individuals who have significant physical disabilities. Over the last ten years, our team in partnership with researchers at the Electrical and Computer Engineering Department, the University of British Columbia, has been working to develop a direct brain-controlled switch for individuals with significant physical disabilities. The NSF Brain Interface Project primarily focuses on the development of brain-computer interface switch technologies for intermittent (or asynchronous) control in natural environments. That is, technologies that will work when the User intends control but also remains in a stable off state when there is no intent to control. A prototype of such a switch has successfully been developed. This switch has demonstrated classification accuracies greater than 94%. The initial results are promising, but further research is required to improve switch accuracies and reliability and to test these switch technologies over a larger population of users and operating conditions. This paper provides an overview of the NSF brain-switch technologies and details our approach to future work in this area. PMID- 12899254 TI - The use of EEG modifications due to motor imagery for brain-computer interfaces. AB - The opening of a communication channel between brain and computer [brain-computer interface (BCI)] is possible by using changes in electroencephalogram (EEG) power spectra related to the imagination of movements. In this paper, we present results obtained by recording EEG during an upper limb motor imagery task in a total of 18 subjects by using low-resolution surface Laplacian, different linear and quadratic classifiers, as well as a variable number of scalp electrodes, from 2 to 26. The results (variable correct classification rate of mental imagery between 75% and 95%) suggest that it is possible to recognize quite reliably ongoing mental movement imagery for BCI applications. PMID- 12899253 TI - Boosting bit rates and error detection for the classification of fast-paced motor commands based on single-trial EEG analysis. AB - Brain-computer interfaces (BCIs) involve two coupled adapting systems--the human subject and the computer. In developing our BCI, our goal was to minimize the need for subject training and to impose the major learning load on the computer. To this end, we use behavioral paradigms that exploit single-trial EEG potentials preceding voluntary finger movements. Here, we report recent results on the basic physiology of such premovement event-related potentials (ERP). 1) We predict the laterality of imminent left- versus right-hand finger movements in a natural keyboard typing condition and demonstrate that a single-trial classification based on the lateralized Bereitschaftspotential (BP) achieves good accuracies even at a pace as fast as 2 taps/s. Results for four out of eight subjects reached a peak information transfer rate of more than 15 b/min; the four other subjects reached 6-10 b/min. 2) We detect cerebral error potentials from single false-response trials in a forced-choice task, reflecting the subject's recognition of an erroneous response. Based on a specifically tailored classification procedure that limits the rate of false positives at, e.g., 2%, the algorithm manages to detect 85% of error trials in seven out of eight subjects. Thus, concatenating a primary single-trial BP-paradigm involving finger classification feedback with such secondary error detection could serve as an efficient online confirmation/correction tool for improvement of bit rates in a future BCI setting. As the present variant of the Berlin BCI is designed to achieve fast classifications in normally behaving subjects, it opens a new perspective for assistance of action control in time-critical behavioral contexts; the potential transfer to paralyzed patients will require further study. PMID- 12899255 TI - EEG changes accompanying learned regulation of 12-Hz EEG activity. AB - We analyzed 15 sessions of 64-channel electroencephalographic (EEG) data recorded from a highly trained subject during sessions in which he attempted to regulate power at 12 Hz over his left- and right-central scalp to control the altitude of a cursor moving toward target boxes placed at the top-, middle-, or bottom-right of a computer screen. We used infomax independent component analysis (ICA) to decompose 64-channel EEG data from trials in which the subject successfully up- or down-regulated the measured EEG signals. Applying time-frequency analysis to the time courses of activity of several of the resulting 64 independent EEG components revealed that successful regulation of the measured activity was accompanied by extensive, asymmetrical changes in power and coherence, at both nearby and distant frequencies, in several parts of cortex. A more complete understanding of these phenomena could help to explain the nature and locus of learned regulation of EEG rhythms and might also suggest ways to further optimize the performance of brain-computer interfaces. PMID- 12899256 TI - A BCI-based environmental controller for the motion-disabled. AB - With the development of brain-computer interface (BCI) technology, researchers are now attempting to put current BCI techniques into practical application. This paper presents an environmental controller using a BCI technique based on steady state visual evoked potential. The system is composed of a stimulator, a digital signal processor, and a trainable infrared remote-controller. The attractive features of this system include noninvasive signal recording, little training requirement, and a high information transfer rate. Our test results have shown that this system can distinguish at least 48 targets and provide a transfer rate up to 68 b/min. The system has been applied to the control of an electric apparatus successfully. PMID- 12899257 TI - Comparison of linear, nonlinear, and feature selection methods for EEG signal classification. AB - The reliable operation of brain-computer interfaces (BCIs) based on spontaneous electroencephalogram (EEG) signals requires accurate classification of multichannel EEG. The design of EEG representations and classifiers for BCI are open research questions whose difficulty stems from the need to extract complex spatial and temporal patterns from noisy multidimensional time series obtained from EEG measurements. The high-dimensional and noisy nature of EEG may limit the advantage of nonlinear classification methods over linear ones. This paper reports the results of a linear (linear discriminant analysis) and two nonlinear classifiers (neural networks and support vector machines) applied to the classification of spontaneous EEG during five mental tasks, showing that nonlinear classifiers produce only slightly better classification results. An approach to feature selection based on genetic algorithms is also presented with preliminary results of application to EEG during finger movement. PMID- 12899258 TI - How many people are able to operate an EEG-based brain-computer interface (BCI)? AB - Ninety-nine healthy people participated in a brain-computer interface (BCI) field study conducted at an exposition held in Graz, Austria. Each subject spent 20-30 min on a two-session BCI investigation. The first session consisted of 40 trials conducted without feedback. Then, a subject-specific classifier was set up to provide the subject with feedback, and the second session--40 trials in which the subject had to control a horizontal bar on a computer screen--was conducted. Subjects were instructed to imagine a right-hand movement or a foot movement after a cue stimulus depending on the direction of an arrow. Bipolar electrodes were mounted over the right-hand representation area and over the foot representation area. Classification results achieved with 1) an adaptive autoregressive model (39 subjects) and 2) band power estimation (60 subjects) are presented. Roughly 93% of the subjects were able to achieve classification accuracy above 60% after two sessions of training. PMID- 12899259 TI - A decision tree for brain-computer interface devices. AB - This paper is a first attempt to present a "decision tree" to assist in choosing a brain-computer interface device for patients who are nearly or completely "locked-in" (cognitively intact but unable to move or communicate.) The first step is to assess any remaining function. There are six inflexion points in the decision-making process. These depend on the functional status of the patient: 1) some residual movement; 2) no movement, but some residual electromyographic (EMG) activity; 3) fully locked-in with no EMG activity or movements but with conjugate eye movements; 4) same as 3 but with disconjugate eye movements; 5) same as 4 but with inadequate assistance from the available EEG-based systems; 6) same as 5 and accepting of an invasive system. PMID- 12899260 TI - Silicon-substrate intracortical microelectrode arrays for long-term recording of neuronal spike activity in cerebral cortex. AB - This study investigated the use of planar, silicon-substrate microelectrodes for chronic unit recording in the cerebral cortex. The 16-channel microelectrodes consisted of four penetrating shanks with four recording sites on each shank. The chronic electrode assembly included an integrated silicon ribbon cable and percutaneous connector. In a consecutive series of six rats, 5/6 (83%) of the implanted microelectrodes recorded neuronal spike activity for more than six weeks, with four of the implants (66%) remaining functional for more than 28 weeks. In each animal, more than 80% of the electrode sites recorded spike activity over sequential recording sessions during the postoperative time period. These results provide a performance baseline to support further electrode system development for intracortical neural implant systems for medical applications. PMID- 12899261 TI - Dynamical dimension of a hybrid neurorobotic system. AB - The goal of this work is to understand how neural tissue can be programed to execute predetermined functions. We developed a research tool that includes the brainstem of a lamprey and a two-wheeled robot interconnected in a closed loop. We report here the development of a framework for studying the dynamics of the neural tissue based on the interaction of this tissue with the robot. PMID- 12899262 TI - Asynchronous BCI and local neural classifiers: an overview of the Adaptive Brain Interface project. AB - In this communication, we give an overview of our work on an asynchronous brain computer interface (where the subject makes self-paced decisions on when to switch from one mental task to the next) that responds every 0.5 s. A local neural classifier tries to recognize three different mental tasks; it may also respond "unknown" for uncertain samples as the classifier has incorporated statistical rejection criteria. We report our experience with 15 subjects. We also briefly describe two brain-actuated applications we have developed: a virtual keyboard and a mobile robot (emulating a motorized wheelchair). PMID- 12899263 TI - Real-world applications for brain-computer interface technology. AB - The mission of the Georgia State University BrainLab is to create and adapt methods of human-computer interaction that will allow brain-computer interface (BCI) technologies to effectively control real-world applications. Most of the existing BCI applications were designed largely for training and demonstration purposes. Our goal is to research ways of transitioning BCI control skills learned in training to real-world scenarios. Our research explores some of the problems and challenges of combining BCI outputs with human-computer interface paradigms in order to achieve optimal interaction. We utilize a variety of application domains to compare and validate BCI interactions, including communication, environmental control, neural prosthetics, and creative expression. The goal of this research is to improve quality of life for those with severe disabilities. PMID- 12899264 TI - Linear and nonlinear methods for brain-computer interfaces. AB - At the recent Second International Meeting on Brain-Computer Interfaces (BCIs) held in June 2002 in Rensselaerville, NY, a formal debate was held on the pros and cons of linear and nonlinear methods in BCI research. Specific examples applying EEG data sets to linear and nonlinear methods are given and an overview of the various pros and cons of each approach is summarized. Overall, it was agreed that simplicity is generally best and, therefore, the use of linear methods is recommended wherever possible. It was also agreed that nonlinear methods in some applications can provide better results, particularly with complex and/or other very large data sets. PMID- 12899265 TI - Training locked-in patients: a challenge for the use of brain-computer interfaces. AB - Training severely paralyzed patients to use a brain-computer interface (BCI) for communication poses a number of issues and problems. Over the past six years, we have trained 11 patients to self-regulate their slow cortical brain potentials and to use this skill to move a cursor on a computer screen. This paper describes our experiences with this patient group including the problems of accepting and rejecting patients, communicating and interacting with patients, how training may be affected by social, familial, and institutional circumstances, and the importance of motivation and available reinforcers. PMID- 12899266 TI - Response error correction--a demonstration of improved human-machine performance using real-time EEG monitoring. AB - We describe a brain-computer interface (BCI) system, which uses a set of adaptive linear preprocessing and classification algorithms for single-trial detection of error related negativity (ERN). We use the detected ERN as an estimate of a subject's perceived error during an alternative forced choice visual discrimination task. The detected ERN is used to correct subject errors. Our initial results show average improvement in subject performance of 21% when errors are automatically corrected via the BCI. We are currently investigating the generalization of the overall approach to other tasks and stimulus paradigms. PMID- 12899267 TI - Graz-BCI: state of the art and clinical applications. AB - The Graz-brain-computer interface (BCI) is a cue-based system using the imagery of motor action as the appropriate mental task. Relevant clinical applications of BCI-based systems for control of a virtual keyboard device and operations of a hand orthosis are reported. Additionally, it is demonstrated how information transfer rates of 17 b/min can be acquired by real time classification of oscillatory activity. PMID- 12899268 TI - Learning to control brain rhythms: making a brain-computer interface possible. AB - The ability to control electroencephalographic rhythms and to map those changes to the actuation of mechanical devices provides the basis for an assistive brain computer interface (BCI). In this study, we investigate the ability of subjects to manipulate the sensorimotor mu rhythm (8-12-Hz oscillations recorded over the motor cortex) in the context of a rich visual representation of the feedback signal. Four subjects were trained for approximately 10 h over the course of five weeks to produce similar or differential mu activity over the two hemispheres in order to control left or right movement in a three-dimensional video game. Analysis of the data showed a steep learning curve for producing differential mu activity during the first six training sessions and leveling off during the final four sessions. In contrast, similar mu activity was easily obtained and maintained throughout all the training sessions. The results suggest that an intentional BCI based on a binary signal is possible. During a realistic, interactive, and motivationally engaging task, subjects learned to control levels of mu activity faster when it involves similar activity in both hemispheres. This suggests that while individual control of each hemisphere is possible, it requires more learning time. PMID- 12899269 TI - A data analysis competition to evaluate machine learning algorithms for use in brain-computer interfaces. AB - We present three datasets that were used to conduct an open competition for evaluating the performance of various machine-learning algorithms used in brain computer interfaces. The datasets were collected for tasks that included: 1) detecting explicit left/right (L/R) button press; 2) predicting imagined L/R button press; and 3) vertical cursor control. A total of ten entries were submitted to the competition, with winning results reported for two of the three datasets. PMID- 12899270 TI - Controlling cellular reactive responses around neural prosthetic devices using peripheral and local intervention strategies. AB - While chronic use of indwelling micromachined neural prosthetic devices has great potential, the development of reactive responses around them results in a decrease in electrode function over time. Since the cellular events responsible for these responses may be anti-inflammatory in nature, we have tested the effectiveness of dexamethasone and cyclosporin A as potential drugs for developing intervention strategies following insertion of single-shank micromachined silicon devices. Peripheral injection of dexamethasone was effective in attenuating increased expression of glial fibrillary acidic protein and astrocyte hyperplasia observed during both initial- and sustained-reactive responses observed at one and six weeks post insertion, respectively. Peripheral injection of cyclosporin A had no positive effect. If anything, application of this drug increased the early reactive response. Effectiveness of local release of dexamethasone in rat neocortex was tested by inserting ribbons of poly (ethyl vinyl) acetate containing 35% (w/w) dexamethasone. Initial concentrations of dexamethasone were similar to those obtained by peripheral injection. Local drug release provided continued control of cellular reactive responses during the six week study period. These results demonstrate that peripheral delivery of dexamethasone can be used to control reactive responses and that local drug delivery by slow-release from biocompatible polymers may be a more effective method of drug intervention. Incorporating these strategies on micromachined devices may provide an intervention strategy that will insure the chronic functioning of electrodes on intracortical neuroprosthetic devices. PMID- 12899271 TI - Data communication between brain implants and computer. AB - Recent advances in neuroscience, microelectronics, and information technology have allowed construction of miniature, but highly intelligent, devices to be implanted within the brain to perform in vitro diagnostic and therapeutic functions. However, there exists a significant problem in establishing an effective wireless data communication link between brain implants and external computer. This communication investigates this link and presents a new design using the mechanism of volume conduction of biological tissues. A theoretical model of volume conduction of the head is utilized to compute signal strength in data communication and the result is evaluated by a physical model. The two-way data communication sensitivity of the volume conduction channel is found to be symmetric, as suggested by the reciprocity theorem. A high-performance, x-shaped volume conduction antenna has been designed. Experiments are performed on animals which demonstrate the effectiveness of this volume conduction approach. PMID- 12899272 TI - Probabilistic methods in BCI research. AB - This paper suggests a probabilistic treatment of the signal processing part of a brain-computer interface (BCI). We suggest two improvements for BCIs that cannot be obtained easily with other data driven approaches. Simply by using one large joint distribution as a model of the entire signal processing part of the BCI, we can obtain predictions that implicitly weight information according to its certainty. Offline experiments reveal that this results in statistically significant higher bit rates. Probabilistic methods are also very useful to obtain adaptive learning algorithms that can cope with nonstationary problems. An experimental evaluation shows that an adaptive BCI outperforms the equivalent static implementations, even when using only a moderate number of trials. This suggests that adaptive translation algorithms might help in cases where brain dynamics change due to learning effects or fatigue. PMID- 12899273 TI - Information conveyed through brain-control: cursor versus robot. AB - Microwire electrode arrays were implanted in the motor and premotor cortical areas of rhesus macaques. The recorded activity was used to control the three dimensional movements of a virtual cursor and of a robotic arm in real time. The goal was to move the cursor or robot to one of eight targets. Average information conveyed about the intended target was calculated from the observed trajectories at 30-ms intervals throughout the movements. Most of the information about intended target was conveyed within the first second of the movement. For the brain-controlled cursor, the instantaneous information transmission rate was at its maximum at the beginning of each movement (averaged 4.8 to 5.5 bits/s depending on the calculation method used). However, this instantaneous rate quickly slowed down as the movement progressed and additional information became redundant. Information was conveyed more slowly through the brain-controlled robot due to the dynamics and noise of the robot system. The brain-controlled cursor data was also used to demonstrate a method for optimizing information transmission rate in the case where repeated cursor movements are used to make long strings of sequential choices such as in a typing task. PMID- 12899274 TI - Multimodal neuroelectric interface development. AB - We are developing electromyographic and electroencephalographic methods, which draw control signals for human-computer interfaces from the human nervous system. We have made progress in four areas: 1) real-time pattern recognition algorithms for decoding sequences of forearm muscle activity associated with control gestures; 2) signal-processing strategies for computer interfaces using electroencephalogram (EEG) signals; 3) a flexible computation framework for neuroelectric interface research; and d) noncontact sensors, which measure electromyogram or EEG signals without resistive contact to the body. PMID- 12899275 TI - The Wadsworth Center brain-computer interface (BCI) research and development program. AB - Brain-computer interface (BCI) research at the Wadsworth Center has focused primarily on using electroencephalogram (EEG) rhythms recorded from the scalp over sensorimotor cortex to control cursor movement in one or two dimensions. Recent and current studies seek to improve the speed and accuracy of this control by improving the selection of signal features and their translation into device commands, by incorporating additional signal features, and by optimizing the adaptive interaction between the user and system. In addition, to facilitate the evaluation, comparison, and combination of alternative BCI methods, we have developed a general-purpose BCI system called BCI-2000 and have made it available to other research groups. Finally, in collaboration with several other groups, we are developing simple BCI applications and are testing their practicality and long-term value for people with severe motor disabilities. PMID- 12899276 TI - Chewing on cardboard and other pleasures of knowledge utilization. PMID- 12899277 TI - A model for interdisciplinary collaboration. AB - Social workers have worked with colleagues from other disciplines since the early days of the profession; yet, they were without clear models to guide this interdisciplinary work. The author uses multidisciplinary theoretical literature and conceptual and research pieces from social work literature to support the development of such a model. First, current trends relevant to interdisciplinary practice are noted to emphasize its importance. The article describes a two-part model. Part one of the model consists of five components that constitute interdisciplinary collaboration between social workers and other professionals: interdependence, newly created professional activities, flexibility, collective ownership of goals, and reflection on process. Part two of the model consists of four influences on collaboration: professional role, structural characteristics, personal characteristics and a history of collaboration. Implications for social work practice are discussed. PMID- 12899278 TI - Interorganizational relationships among nonprofits in the aftermath of welfare reform. AB - Although it is widely acknowledged that nonprofit agencies are critical actors in the nation's social service arena, few studies to date have investigated how the 1996 welfare reform legislation affected them. This article reports on one component of a research project in southeast Michigan that explored how the Personal Responsibility and Work Opportunity Reconciliation Act of 1996 transformed the interorganizational environment of private nonprofit social services providers. The study found that smaller agencies and those that focus on clients' emergency needs appeared to be particularly vulnerable in today's climate. Private nonprofit providers, however, are critical to the survival of low-income individuals and families, particularly TANF recipients, and to the ultimate success of welfare reform. PMID- 12899279 TI - Advocating in schools for children with disabilities: what's new with IDEA? AB - All social workers who work with children and families, regardless of their practice setting, should be aware of the important educational rights to which children with disabilities and their families are entitled. The Individuals with Disabilities Education Act (P.L. 101-476), one of the most sweeping laws protecting children with disabilities, was substantially amended in 1997, and its regulatory provisions became legally effective in October 1999. This article provides information about the requirements of the law and the impact of those changes on children's educational rights. The changes discussed and their practice implications include expansion of categories of children with disabilities; new requirements for mobile, homeless, or culturally diverse populations and participants in the individualized education program process; payment for private school placements for children with disabilities; discipline of children with disabilities; and provision of social work services in the schools. PMID- 12899280 TI - People with cognitive disabilities: the Argument from Marginal Cases and social work ethics. AB - A primary argument used by animal rights advocates to engage the public in questioning our maltreatment of animals is the Argument from Marginal Cases (AMC). It is important that social workers are aware of this argument, because it has the potential to diminish our consideration of people with severe cognitive disabilities. This article provides a brief overview of the argument, followed by a description of the means by which people with cognitive disabilities have been denigrated over the past century through animalistic rhetoric and negative comparisons with animals. The "animalization" of marginalized groups of all types has often served to reinforce and justify dehumanizing treatment of group members. Questions related to the logic of the AMC are raised, and the importance of the argument for the social work community is discussed. PMID- 12899281 TI - Social work and the law: a therapeutic jurisprudence perspective. AB - Social workers in all practice areas are affected by interactions with the law and legal systems. Unfortunately, the profession has paid inadequate attention to education, scholarship, and practice in legal environments. This article introduces a new perspective for examining social work and the law called "therapeutic jurisprudence," which emanates from psychology/mental health law. Therapeutic jurisprudence focuses on the therapeutic and antitherapeutic consequences of laws, legal rules, and legal actions. It provides a framework for assessing legal issues, applying social science research to legal questions, and structuring interventions in the legal environment. The authors argue that this perspective has the potential to serve as a lens through which social workers can view the law and, ultimately, use the knowledge gained to achieve better outcomes for clients. PMID- 12899282 TI - Probation foster care as an outcome for children exiting child welfare foster care. AB - Little research has been done to help us understand what happens to children who exit child welfare foster care for reasons other than emancipation. Almost no research exists to help us understand out-of-home placement supervised by other providers of children's social services such as juvenile probation or mental health. This study examined school-age children who entered out-of-home placement supervised by probation departments after they left child welfare foster or group care. Instability in child welfare placements significantly increased the risk of a transition to probation foster care. Among the children exiting child welfare placements, those who had entered their first spell in care at ages 12 through 14 or were first removed because of sexual abuse or neglect were at greater risk of probation out-of-home placement. Implications for social work policy and services are discussed. PMID- 12899283 TI - From misery to mission: forensic social workers on multidisciplinary mitigation teams. AB - Social workers are well-equipped by experience and education to play a pivotal role in death penalty mitigation teams. They offer expertise in researching complete social histories, providing for people under threat of execution, and helping those individuals cope with that threat. The social worker's primary role is to develop the client's story through an extensive empirical inquiry into the person's life. An equally important role for social workers is to work with the multidisciplinary defense team to ensure that the client's life story becomes a part of the defense. This article, through a case example of a condemned prisoner, examines the mitigation team concept, focusing on the social work role. PMID- 12899284 TI - What is Catholic about Catholic Charities? AB - Sectarian social services agencies play an important and increasing role in contemporary social welfare. Among sectarian social welfare organizations, Catholic Charities USA has emerged as the largest private provider of social welfare services. This article reviews the history, services, and practice controversies of Catholic Charities USA and examines issues regarding the ability of sectarian social services organizations to provide nonbiased and fair services. Through an analysis of this organization, the authors raise and discuss questions of accountability and philosophical approaches. PMID- 12899285 TI - Managing managed care through accreditation standards. AB - Accreditation is emerging as a dominant strategy for ensuring accountability and basic quality of services in managed care organizations. Knowledge of accreditation standards can help social workers interact effectively with managed care companies when requesting services for clients and appealing denials. Familiarity with patient's rights protections, the appeals process, and provider credentialing guidelines can help protect social workers as well as clients from unfair decisions by managed care organizations. This article provides an overview of managed care accreditation standards and strategies for integrating this knowledge into advocacy efforts. PMID- 12899286 TI - Finding meaning in medical necessity. AB - Managed care organizations have become partners in patient care through their representatives/liaisons---care managers. This article uses personal experiences and relevant literature to highlight the professional conflicts care managers face in the struggle to fulfill two main clinical functions: applying criteria to determine medical necessity and educating practitioners and patients about effective and efficient treatments. Many care managers struggle to maintain cordial relationships with colleagues, while determining which requests for treatment are necessary versus discretionary, and insuring treatment excellence. PMID- 12899287 TI - Excavating our frames of mind: the key to dialogue and collaboration. AB - Conversations between colleagues who hold divergent views of a case or practice situation often take the form of a win-lose debate in which contenders, arguing from within their respective frameworks, find they are unable to reach mutual understanding. The resulting impasse makes mutual understanding and collaboration difficult. Our typical way of resolving the impasse is to invoke the evidence for our own position, hoping that the facts somehow speak for themselves. Strangely, this may only exacerbate the impasse and worsen our view of our colleagues, whom we presume to be ignoring the facts. This article attempts to show that reflecting on the frame--not debating the facts--might be a more helpful way to approach such impasses and to reach mutual understanding. PMID- 12899288 TI - Open adoption of infants: adoptive parents' feelings seven years later. AB - Adoptions today increasingly include contact between adoptive and birth families. What do these "open adoptions" look like? How do the participants feel about them? This article, based on part of a longitudinal study that first examined adoptive parents' perceptions of their infants' open adoptions seven years ago, explores the parents' reactions now that their children are school age. This qualitative descriptive research revealed changes in the openness in the adoptions over time and identified four dimensions along which open adoptions vary. Findings showed parents' enthusiasm for the openness in their adoptions, regardless of the type and extent of openness. Implications for social work practice, education, and policy are explored. PMID- 12899289 TI - Enhancing relationships in nursing homes through empowerment. PMID- 12899290 TI - When is a mental health clinic not a mental health clinic? Drug trial abuses reach social work. PMID- 12899291 TI - Social work, Evangelical Christians, and values. PMID- 12899292 TI - Comparison of methods used for pre-concentrating small volumes of organic volatile solutions. AB - Eight pre-concentration techniques were compared for their capacity to retain volatile and semi-volatile solutes during evaporation of solvent (dichloromethane). The 2-ml test-samples containing 0.2 ppm or 2 ppm (v/v) of volatile and semi-volatile solutes were concentrated to a final volume of 1 ml, 200 microl and 50 microl, respectively. When pre-concentrating to 50 microl, the highest recoveries for both the diluted (0.2 ppm) and concentrated (2 ppm) solutions were found by passive evaporation in a test tube at 22 degrees C. The pre-concentration time from 2 ml to 50 microl by this method was 19-20 h. Heating the test tube to 47 degrees C yielded lower recoveries in dilute samples, but the recoveries of concentrated samples were only slightly lower than the recoveries obtained by passive evaporation. The evaporation time was decreased to 1-2 h. The recoveries and the reproducibility of these methods were superior to the other pre-concentration methods tested. Loss of solute was apparently mainly caused by the fast vapour streams created when speeding up the process of evaporation by heating or by introducing a gas stream into the tube. This increased co evaporation and thereby solute loss. The capacity of the methods to trap the escaping vapours and create a reflux determined the capacity of the methods to recover the solutes. The experiments demonstrated that more solute is lost during the pre-concentration of dilute samples compared to more concentrated solutions. PMID- 12899293 TI - On-chip protein sample desalting and preparation for direct coupling with electrospray ionization mass spectrometry. AB - A membrane-based desalting step integrated in a MS microchip is presented: drugs, peptides and proteins are adsorbed on a hydrophobic poly(vinylidene difluoride) membrane, which allows the washing out of salts. The integration with microfluidics permits a controlled elution of analytes from the membrane and their direct mass spectrometric analysis by electrospray ionisation MS. The desalting process is demonstrated with picomole amounts of propanolol, insulin and cytochrome c. Moreover, this stop-and-go desalting process is tolerant to high concentrations of urea, and to the presence of reductants such as dithiothreitol. This particular feature allowed the chemical tagging of cysteines in beta-lactoglobulin A with iodoacetamide. Finally, the integration of chemical tagging, on-chip desalting and MS microchip paves the way for the development of high-throughput analytical procedure for structural proteomics. PMID- 12899294 TI - Determination of antibiotics from soil by pressurized liquid extraction and liquid chromatography-tandem mass spectrometry. AB - A method for the analysis of several macrolide and ionophore antibiotics, as well as tiamulin, from soil was developed using pressurized liquid extraction (PLE), reversed-phase liquid chromatography and atmospheric pressure chemical ionisation (APCI) tandem mass spectrometry (LC-APCI+-MS-MS). The analytes were extracted from soil by PLE in 30 min and the extracts were cleaned up by solid-phase extraction (SPE) on a diol SPE cartridge. Liquid chromatographic (LC) separation of the antibiotics was achieved in 35 min. Recovery experiments were performed using spiked soil and concentrations varying from 1 to 2000 microg/kg. By using a macrolide internal standard the recovery rates for the macrolides erythromycin and roxithromycin ranged from 43 to 94% (RSD 20-23%), for the ionophore salinomycin the recovery rate was 76% (RSD 29%), while the pleuromutilin tiamulin was completely recovered. The limits of detection ranged from 0.2 to 1.6 microg/kg. In soil samples a maximum concentration of 0.7 microg/kg tiamulin was found. PMID- 12899295 TI - Solid-phase extraction clean-up of soil and sediment extracts for the determination of various types of pollutants in a single run. AB - A new sample clean-up procedure based on solid-phase extraction (SPE) sorbents was proposed for the determination of pesticides, polycyclic aromatic hydrocarbons and polychlorinated biphenyls in soils and sediments. The main purpose of the research was to find a combination of sorbents for the SPE method that would permit the determination of many types of analytes (polycyclic aromatic hydrocarbons, polychlorinated biphenyls, N-, P- and Cl-containing pesticides) in a single run. Elution profiles for both the analytes and the interfering components were determined for several types of SPE sorbents (alumina, silica and surface-modified silica) and combinations of them. The efficiency of the clean-up method developed was evaluated using real soil samples. PMID- 12899296 TI - Repeatability and reproducibility of high concentration data in reversed-phase liquid chromatography. I. Overloaded band profiles on Kromasil-C18. AB - Single-component adsorption-isotherm data were acquired by frontal analysis (FA) for six low-molecular-mass compounds (phenol, aniline, caffeine, theophylline, ethylbenzene and propranolol) on one Kromasil-C18 column, using water-methanol solutions (between 70:30 and 20:80, v/v) as the mobile phase. Propranolol data were also acquired using an acetate buffer (0.2 M) instead of water. The data were modeled for best agreement between calculated and experimental overloaded band profiles. The adsorption energy distribution was also derived and used for the selection of the best isotherm model. Widely different isotherm models were found to model best the data obtained for these compounds, convex upward (i.e. Langmuirian), convex downward (i.e. anti-Langmuirian), and S-shaped isotherms. Using the same sample size for all columns (loading factor, Lf approximately 10%), overloaded band profiles were recorded on four different columns packed with the same batch of Kromasil-C18 and five other columns packed with different batches of Kromasil-C18. These experimental band profiles were compared to the profile calculated from the isotherm measured by FA on the first column. The repeatability as well as the column-to-column and the batch-to-batch reproducibilities of the band profiles are better than 4%. PMID- 12899297 TI - Study of the competitive isotherm model and the mass transfer kinetics for a BET binary system. AB - The competitive adsorption behavior of the binary mixture of phenetole (ethoxy benzene) and propyl benzoate in a reversed-phase system was investigated. The adsorption equilibrium data of the single-component systems were acquired by frontal analysis. The same data for binary mixtures were acquired by the perturbation method. For both compounds, the single-component isotherm data fit best to the multilayer BET model. The experimental overloaded band profiles are in excellent agreement with the profiles calculated with either the general rate model or the modified transport-dispersive models. The competitive adsorption data were modeled using the ideal adsorbed solution (IAS) theory. The numerical values of the coefficients were derived by fitting the retention times of the perturbation pulses to those calculated using the IAS theory compiled with the coherence conditions. Finally, the elution profiles of binary mixtures were recorded. They compared very well with those calculated. As a characteristic feature of this case, an unusual retainment effect of the chromatographic band of the more retained component by the less retained one was observed. The combination of the General Rate Model and the adsorption isotherm model allowed an accurate prediction of the band profiles. PMID- 12899298 TI - Analytical solution of the ideal, nonlinear model of reaction chromatography for a reaction A-->B and a parabolic isotherm. AB - The analytical solution of the ideal, nonlinear model of reaction chromatography for the simplest possible chemical reaction, A-->B and with a parabolic isotherm for the reagent, is derived for two types of boundary conditions, the injection of a rectangular concentration pulse of finite width (elution) and that of an instantaneous concentration jump (Riemann problem or breakthrough curve). The areas of the profiles of the reagent and of the product of the reaction are calculated in both the ideal and the nonideal cases. The effects of the nonlinear behavior of the equilibrium isotherm and of axial dispersion on the reagent profile are discussed using analytical and numerical methods. PMID- 12899299 TI - Effect of phase ratio on van't Hoff analysis in reversed-phase liquid chromatography, and phase-ratio-independent estimation of transfer enthalpy. AB - In analysis of the thermodynamics of the transfer of a solute from the mobile phase to the stationary phase in reversed-phase liquid chromatography, it is nearly always assumed that the phase ratio is constant. This type of analysis is typically performed by applying a form of the van't Hoff equation, which relates the retention factor to temperature via the enthalpy and entropy of transfer. When non-linear van't Hoff plots are observed, it is often assumed that the enthalpy and entropy of transfer change with temperature. However, when the possibility of a change in the phase ratio is considered, it becomes apparent that non-linear van't Hoff behavior may or may not be due to changes in enthalpy or entropy. In this work, we present mathematical evidence that phase ratio changes, if they occur, can cause deviations from linearity in a van't Hoff plot. We also show that the phase ratio influence can be eliminated by considering the molecular difference between two solutes instead of the solutes themselves. The resulting selectivity van't Hoff plots may be linear, even when the van't Hoff plots of the two solutes are non-linear. In such cases, temperature-dependent phase ratio changes, and not necessarily changes in the transfer enthalpy, may be responsible for the curved van't Hoff plots of the individual solutes. In addition, we present chromatographic evidence that different solutes may "see" different thermodynamic phase ratios. It is clear that the concept of a phase ratio in reversed-phase chromatography is not nearly as well defined as a phase ratio in a bulk system like a liquid-liquid extraction. PMID- 12899300 TI - System maps for retention of neutral organic compounds under isocratic conditions on a reversed-phase monolithic column. AB - The solvation parameter model is used to create systems maps for the separation of neutral organic compounds on a Chromolith Performance RP-18e octadecylsiloxane bonded silica-based monolithic column for water-acetonitrile and water-methanol mobile phase compositions from 10 to 70% (v/v) organic solvent. These results demonstrate that the retention properties of the monolithic column are similar to those of conventional octadecylsiloxane-bonded silica particle-packed columns. It is further shown that the selectivity for the monolithic column falls within the selectivity range for typical particle-packed columns at two mobile phase compositions for which a direct comparison is possible. PMID- 12899301 TI - Isocratic separation of monosaccharides using immobilized Concanavalin A. AB - The successful fractionation of a mixture of monosaccharides using an immobilized lectin column operating under isocratic conditions is described. The key factors for effective separation were found to be sample size, feed velocity and column length. Under optimal conditions it was possible to obtain complete resolution of a mixture of L-arabinose, D-fructose and D-mannose in less than 40 min. PMID- 12899302 TI - Determination of noradrenaline and dopamine in Chinese herbal extracts from Portulaca oleracea L. by high-performance liquid chromatography. AB - A simple, rapid and accurate high-performance liquid chromatographic (HPLC) technique coupled with photodiode array (PDA) detection was developed for the simultaneous determination of noradrenaline (NA) and dopamine (DA) in Chinese herbal plant extracts from the different parts of Portulaca oleracea L. The effects of various parameters, such as sodium dodecyl sulfate (SDS) concentration, pH value and proportion of methanol on chromatographic behavior of the analytes (NA and DA) were investigated. Separation of NA and DA was achieved within 10 min by a mobile phase consisting of 70% (v/v) methanol and 0.02 M potassium dihydrogen phosphate solution, which contained 30 mM SDS and was adjusted to pH 3.0 with H3PO4. NA and DA showed good linear relationships in the range of 0.004-6.00 microg and 0.011-8.25 microg respectively. The correlation coefficients of the calibration curve for the analytes exceeded 0.999. The detection limits for NA and DA were 0.40 ng and 0.55 ng, at a signal-to-noise ratio of 3, respectively. Moreover, the optimized HPLC method was employed to analyze three different parts of Portulaca oleracea L. PMID- 12899303 TI - Determination of nutrients in the presence of high chloride concentrations by column-switching ion chromatography. AB - Determination of inorganic anions in waters of high salinity is one of the most difficult task in analytical chemistry. A simple column-switching method, based on an original chromatographic set-up, for the determination of nutrients (nitrate, nitrite and phosphate) in chloride rich aqueous matrices is presented. A pre-separation system (made of two in line pre-columns, Dionex AG9-HC 4 mm) connected to an analytical column (Dionex AS9-HC 4 mm) by a four way pneumatic valve, allows chloride to be eluted off into the waste and nutrients to be separated and detected by a conductimeter and/or a UV spectrophotometer. Neither chemical pre-treatment nor sample dilution are required; sample matrices presenting a large range of chloride concentrations can be investigated. Moreover by using this technology, automation for routine analysis, low analysis time and low costs can be achieved. LODs of 100, 300, 1000 microg/l for nitrate, nitrite and phosphate, respectively, have been obtained by spiking a synthetic sea water sample containing 20,000 mg/l of chloride and 3000 mg/l of sulphate. Analyte calibration curves of analytes are linear (r>0.99) in the range between the LODs and 60 mg/l. This method was applied to nutrients determination in sea water samples collected near a river outlet. PMID- 12899304 TI - Gas chromatographic-mass spectrometric characterisation of amiton and the recovery of amiton from concrete, paint, rubber and soil matrices. AB - Amiton [O,O-diethyl S-[2-(diethylamino)ethyl] phosphorothiolate], is an organophosphorus chemical included in Schedule 2 of the Chemical Weapons Convention (CWC). Verification provisions under the CWC rely on the existence of a database of analytical information for scheduled chemicals and related compounds. Little analytical information is available for amiton. In this study, gas chromatography-mass spectrometry (GC-MS) characterisation of amiton and its typical impurities (including by-products and degradation products), supported by selective GC detection and 31P NMR data, was undertaken. Twenty-one compounds, including a by-product unique to amiton from an industrial source, were identified. Involatile degradation products of amiton were derivatised to enable their identification by GC-MS. The recovery of amiton from matrices that may be expected in an inspection scenario (i.e. concrete, paint, rubber and soil) was also examined. Paint and concrete matrices were the most useful matrices for the detection of amiton, and its by-products and degradation products. Amiton was readily detected in these matrices after 28 days. PMID- 12899305 TI - Rapid method development for chiral separation in drug discovery using sample pooling and supercritical fluid chromatography-mass spectrometry. AB - A novel strategy for rapid chiral method development has been implemented using sample pooling and supercritical fluid chromatography-mass spectrometry (SFC-MS) on four chiral stationary phases, namely Chiralpak AD and AS, and Chiralcel OJ and OD, and eight different modifier concentrations (5 to 40% methanol-0.2% isopropylamine). The screening is performed under an outlet pressure of 110 bar at 35 degrees C, and at a flow-rate of 2.5 ml/min for the initial 20 min and then ramped up to 4 ml/min and held for 4.5 min to elute all solutes from the column. The entire process is fully automated from injection to data processing, and operates unattended for 15 h overnight to obtain optimal chiral separation for multiple compounds. A unique feature of using SFC-MS to monitor chiral synthesis is the negligible interferences from achiral impurities. In addition, with SFC MS, enantiomeric excess can be determined with much lower detection limits than UV and much shorter analysis times compared to normal-phase/reversed-phase liquid chromatography. PMID- 12899306 TI - Chiral analysis of neurotransmitters using cyclodextrin-modified capillary electrophoresis equipped with microfabricated interdigitated electrodes. AB - We present cyclodextrin-modified capillary electrophoresis equipped with a microfabricated chip consisting of an array of eight interdigitated microband platinum electrodes (IDs) for simultaneous analysis of three chiral models: epinephrine, norepinephrine and isoproterenol. The IDE chip, positioned very close to the capillary outlet, served as an amplification/detection system. Emerging neurotransmitters at the IDE surface were oxidized at +1.1 V by seven electrodes of the array and then detected by the remaining electrode, poised at +0.0 V. There was an amplification effect on the detecting electrode owing to the recycle between the reduced and oxidized forms of the optical isomers at the electrode surface. The detecting "amplification" current response was governed by the applied potential, the detecting electrode position, the number of adjacent electrodes used for recycling and the distance between the oxidative and reductive electrodes. The six chiral forms of the three neurotransmitters were resolved using 25 mM heptakis(2,6,di-o-methyl)-beta-cyclodextrin with a detection limit of approximately 5 microM. The scheme detected a reduced compound at a reducing potential instead of conventional oxidation detection to alleviate electrode fouling and electroactive interferences. The concurrent oxidation/reduction detection of compounds also facilitated and ascertained peak identification as interfering compounds were unlikely to have the same oxidative/reductive characteristics and mobilities as the analytes of interrogation. PMID- 12899307 TI - Capillary electrophoretic analysis of the derivatives and isomers of benzoate and phthalate. AB - A capillary electrophoretic method for the analysis of 12 commonly found derivatives and isomers of benzoate and phthalate, including p-toluic acid, p acetamido and p-hydroxy derivatives of benzoic acid, salicylic acid and its acetyl ester, 2- and 4-isomers of carboxybenzaldehyde, meta-, para-, and ortho isomers of phthalic acid, and monomethyl terephthalic acid was developed. Capillary electrophoresis (CE) was performed in the free zone electrophoresis mode. Performing CE in 10 mM phosphate buffer, pH 7.0 could separate most of the benzoic acid derivatives except the structural or positional isomers. The positional isomers of phthalic acids could be completely separated with co addition of alpha- and beta-cyclodextrins. Addition of poly(ethylene glycol) 600 (4%) could further resolve some structural isomers. The CE method developed here is rapid, i.e. complete separation could be achieved in less than 8 min for the nine monoanionic benzoate derivatives and in less than 14 min for the three dianionic phthalate isomers. The new method has good precision and linearity and can be readily applied to real samples for quantitative analysis. It is sensitive and can detect sub-ppm (w/w) level of impurity in real terephthalic samples. PMID- 12899308 TI - Determination of imidacloprid and its metabolite 6-chloronicotinic acid in greenhouse air by application of micellar electrokinetic capillary chromatography with solid-phase extraction. AB - A method is described for the analysis of the insecticide imidacloprid [1-(6 chloro-3-pyridylmethyl)-N-nitroimidazolidin-2-ylideneamine] and its metabolite 6 chloronicotinic acid by micellar electrokinetic chromatography with diode-array detection at 270 and 227 nm, respectively. The best results were obtained using sodium dodecyl sulphate at a concentration of 60 mM and a running buffer of NH4Cl/NH3 at 15 mM (pH 8.5). The selection of instrumental parameters such as voltage at 30 kV with an injection time of 20 s gave the best resolution with an analysis time of less than 6 min. The method yields similar sensitivity for the parent compound and for the metabolite, with detection limits of 0.71 and 1.18 microg/ml for imidacloprid and 6-chloronicotinic acid, respectively. The sampling and analysis of these two pesticides in greenhouse air was carried out using personal samplers connected to XAD-2 cartridges as sampling media, investigating the dissipation of analytes in a 24-h period after their application. PMID- 12899309 TI - Simultaneous determination of inorganic nitrogen species by microcolumn ion chromatography. AB - Inorganic nitrogen species (nitrate, nitrite and ammonium ions) were simultaneously determined by microcolumn ion chromatography. Nitrate and nitrite were determined by UV detection at 206 nm, whereas ammonium ion was determined by fluorescence detection at excitation 410 nm and emission 470 nm. The latter fluorescence detection is based on the postcolumn reaction of ammonium ion with o phthalaldehyde in the presence of 2-mercaptoethanol. Effects of the reagent concentration, pH, and other reaction conditions on the signal intensity were examined, and the optimum condition was explored. The present method allowed simultaneous determination of nitrate, nitrite and ammonium ions in river water. PMID- 12899310 TI - Evaluation of multi-walled carbon nanotubes as gas chromatographic column packing. AB - Purified multi-walled carbon nanotubes (PMWCNTs), activated charcoal and graphitized carbon black (Carbopack B) were used as column packing materials to compare their separation ability and other gas chromatographic behavior for aromatic hydrocarbons, alkanes, halogenated hydrocarbons, alcohols, ketones, esters, and ethers. The results show that PMWCNTs can be an excellent gas chromatographic packing material. Compared to Carbopack B with the same surface area, PMWCNTs had a stronger retention ability, a more homogenous surface and smaller theoretical plate numbers. Polar compounds can present symmetric peaks on PMWCNTs. PMWCNTs were found to be an interesting alternative adsorbent to activated charcoal and Carbopack B as gas chromatographic column packing material for volatile compounds, especially those with relatively low boiling points. PMID- 12899311 TI - Separation of biogenic amines by micellar electrokinetic chromatography with on line chemiluminescence detection. AB - A method of on-line chemiluminescence detection with capillary electrophoresis for biogenic amines (diaminopropane, putrescine, cadaverine and diaminohexane) labeled with N-(4-aminobutyl)-N-ethylisoluminol is reported for the first time. Two separation modes, capillary zone electrophoresis and micellar electrokinetic chromatography (MEKC), were studied. The results show that excellent resolution was achieved in MEKC. Parameters affecting separation process and chemiluminescence detection have been examined in detail. Under the optimum conditions, the baseline separation of four amines was obtained within 7.5 min. The detection limits (S/N=3) of diaminopropane, putrescine, cadaverine and diaminohexane are 3.5 x 10(-8), 3.5 x 10(-8), 3.9 x 10(-8) and 1.2 x 10(-7) M, respectively. The method was applied to the analysis of biogenic amines in lake water. PMID- 12899312 TI - Novel polyamine coating providing non-covalent deactivation and reversed electroosmotic flow of fused-silica capillaries for capillary electrophoresis. AB - A new polycationic coating for use in capillary electrophoresis has been developed that enables chemical modification of fused-silica capillary surfaces for analysis of compounds like basic proteins. The cationic polyamine, containing short aliphatic blocks of combined 2 and 3-carbon length, was physically adsorbed onto the negatively charged fused-silica surface through ionic interaction by flushing the capillary with a polyamine solution, followed by a self stabilization step. The polyamine coated capillaries generated an anodal electroosmotic flow that was independent of pH in the investigated range of pH 4 8. The capillary performance was demonstrated by fast separations of basic proteins with peak efficiencies in the range of 265,000-584,000 plates. PMID- 12899313 TI - Individual differences in the responses of naive rhesus monkeys to snakes. AB - The authors demonstrated individual differences in inhibited behavior and withdrawal responses of laboratory-born rhesus monkeys when initially exposed to a snake. Most monkeys displayed a small significant increase in their behavioral inhibition in the presence of a snake. A few monkeys had marked responses, and some actively withdrew. Although the responses of the most extreme laboratory born monkeys were comparable to feral-born monkeys, the responses of the laboratory-born monkeys rapidly habituated. The individual differences in the responses of naive monkeys likely reflect a continuum from orienting to wariness to fear. A neurobiological model is presented that addresses potential mechanisms underlying these individual differences, their relation to fear, and how they may predispose to phobia development. PMID- 12899314 TI - There is more to anger coping than "in" or "out". AB - There is growing dissatisfaction with a dichotomized "anger-in" versus "anger out" view of anger coping. Three studies using student and community adultsamples revealed a broader understanding of the nature of anger coping styles and led to the development of the new Behavioral Anger Response Questionnaire (BARQ). The BARQ is empirically derived and factorially validated and has good psychometrics. Results suggest that dichotomizing anger responses as "in" versus "out" is too coarse and that a 6-factor model may be more appropriate. The 6 factors identified here are Direct Anger-Out, Assertion, Support-Seeking, Diffusion, Avoidance, and Rumination. Women reported use of a wider range of anger coping styles, especially more social support-seeking and more use of anger diffusion strategies than men. PMID- 12899315 TI - Emotionality, emotion regulation, and adaptation among 5- to 8-year-old children. AB - This study investigated relations between emotionality, emotion regulation, and children's behavioral adaptation in a longitudinal design. Mothers rated emotionality and emotion regulation related to anger, fear, and positive emotions exuberance for 151 children at age 5 and later at age 6 years 6 months. Emotionality and emotion regulation measures were modestly related. Preschool ratings at age 6 (n = 125), maternal ratings at age 6 years 6 months (n = 133), and elementary school ratings at age 8 (n = 135) of problems and competence were also collected. High anger emotionality and low regulation of positive emotions and exuberance predicted externalizing problem behavior and prosocial behavior. High fear emotionality and low fear regulation predicted internalizing problem behavior. There were few interactive effects of emotionality and regulation. PMID- 12899316 TI - The social consequences of expressive suppression. AB - At times, people keep their emotions from showing during social interactions. The authors' analysis suggests that such expressive suppression should disrupt communication and increase stress levels. To test this hypothesis, the authors conducted 2 studies in which unacquainted pairs of women discussed an upsetting topic. In Study 1, one member of each pair was randomly assigned to (a) suppress her emotional behavior, (b) respond naturally, or (c) cognitively reappraise in a way that reduced emotional responding. Suppression alone disrupted communication and magnified blood pressure responses in the suppressors' partners. In Study 2, suppression had a negative impact on the regulators' emotional experience and increased blood pressure in both regulators and their partners. Suppression also reduced rapport and inhibited relationship formation. PMID- 12899317 TI - High frequency of facial expressions corresponding to confusion, concentration, and worry in an analysis of naturally occurring facial expressions of Americans. AB - College students were instructed to observe symmetric and asymmetric facial expressions and to report the target's judgment of the "emotion" she or he was expressing, the facial movements involved, and the more expressive side. For both asymmetric and symmetric expressions, some of the most common emotions or states reported are neither included in standard taxonomies of emotion nor studied as important signals. Confusion is the most common descriptor reported for asymmetric expressions and is commonly reported for symmetrical expressions as well. Other frequent descriptors were think-concentrate and worry. Confusion is characterized principally by facial movements around the eyes and has many properties usually attributed to emotions. There was no evidence for lateralization of positive versus negative valenced states. PMID- 12899318 TI - Now you see it, now you don't--the confusing case of confusion as an emotion: commentary on Rozin and Cohen (2003). AB - P. Rozin and A. B. Cohen (2003) contend that confusion is an emotion because it is valenced, it has a distinct facial expression, and it has a distinct internal state. On the basis of these criteria, they call for further study of this unstudied stateand challenge emotion researchers to consider "confusion" to be an emotion. The author agrees with Rozin and Cohen (2003) that confusion is an affective state, is valenced, has an (internal) object, may be expressed facially, and that laypersons may, under certain circumstances, consider it an emotion. However, its expression is likely to be an expressive component of emotions for which goal obstruction is central. Further, confusion may also not be as commonly considered an emotion by laypersons, as Rozin and Cohen contend. Finally, confusion is not unstudied, only most of the time it is not emotion researchers who do the researching. PMID- 12899319 TI - Confusion, concentration, and other emotions of interest: commentary on Rozin and Cohen (2003). AB - P. Rozin and A. B. Cohen's (2003) method of sending students out to observe each other in familiar circumstances undoubtedly exaggerated the apparent prevalence of confusion, concentration, and worry. The expressions they observed probably ranged from regulatory feedback and communicative signals to expressions of the "intellectual emotions" described by C. Darwin (1872/1965). Appraisal theories can easily accommodate these affective states; there is no need to postulate new "basic emotions" unless one adheres to a rigid categorical view of emotion. Finally, Rozin and Cohen have made a valuable contribution by reminding us of the importance of emotions related to interest. PMID- 12899320 TI - New displays and new emotions: a commentary on Rozin and Cohen (2003). AB - In this article, the authors elaborate on 3 ideas advanced in P. Rozin and A. B. Cohen's (2003) innovative study of facial expression. Taking a cue from their discovery of new expressive behaviors (e.g., the narrowed eyebrows), the authors review recent studies showing that emotions are conveyed in more channels than usually studied, including posture, gaze patterns, voice, and touch. Building on their claim that confusion has a distinct display, the authors review evidence showing distinct displays for 3 self-conscious emotions (embarrassment, shame, and pride), 5 positive emotions (amusement, desire, happiness, love, interest), and sympathy and compassion. Finally, the authors offer a functional definition of emotion to integrate these findings on "new" displays and emotions. PMID- 12899321 TI - Measuring emotional intelligence with the MSCEIT V2.0. AB - Does a recently introduced ability scale adequately measure emotional intelligence (EI) skills? Using the Mayer-Salovey-Caruso Emotional Intelligence Test (MSCEIT; J. D. Mayer, P. Salovey, & D. R. Caruso, 2002b), the authors examined (a) whether members of a general standardization sample and emotions experts identified the same test answers as correct, (b) the test's reliability, and (c) the possible factor structures of EI. Twenty-one emotions experts endorsed many of the same answers, as did 2,112 members of the standardization sample, and exhibited superior agreement, particularly when research provides clearer answers to test questions (e.g., emotional perception in faces). The MSCEIT achieved reasonable reliability, and confirmatory factor analysis supported theoretical models of EI. These findings help clarify issues raised in earlier articles published in Emotion. PMID- 12899322 TI - Interleukin 10 inhibits the rat VSMC proliferation and collagen secretion stimulated by angiotensin II. AB - OBJECTIVE: To study the effect of interleukin 10 (IL-10) on the angiotensin II (AngII) stimulated rat VSMC proliferation and collagen secretion, and furthermore, explore its mechanism. METHODS: On cultured VSMC of rat, 3H-thymine (3H-TdR) and 3H-proline incorporations were used to evaluate the DNA and collagen synthesis, respectively. Western blot and immunoprecipitation were applied to assay the expression and activity of focal adhesion kinase (FAK), respectively. RESULTS: IL-10 (10(-8) approximately 10(-10) g/ml) inhibited the increase of 3H TdR and 3H-proline incorporation as well as FAK activity, which was induced by 10(-7) mol/L AngII (P < 0.05 or P < 0.01). IL-10 also obviously downregulated the synthesis and secretion of collagen by AngII stimulated VSMC. But there was no difference in the protein expression of FAK among all the groups (P > 0.05). CONCLUSION: IL-10 antagonizes the VSMC proliferation and collagen synthesis by regulating FAK activity stimulated by AngII. PMID- 12899323 TI - Expression and glycosylation of rotavirus strain SA11 VP4 protein in a recombinant adenovirus. AB - OBJECTIVE: Using a recombinant human adenovirus to express modified VP4 gene of rotavirus SA11 strain. METHODS: A whole VP4 gene was obtained with PCR and induced the signal peptide at the gene N terminal. The chimera gene was cloned into pCMV plasmid that consists of human cytomegalovirus promoter, and then the gene was cloned to the transfer vector of human adenovirus type 5. Homologous recombination was performed by co-transfection to 293 cell lines with recombinant plasmid and viral genome using CaPO4 precipitation. RESULTS: No mutation was found in the whole VP4 gene sequence of 2362 base pair. The expressed product in recombinant adenovirus was confirmed to be specific and more antigenicity by indirect immunofluorescence assay. Both the Western blot and immunoprecipitation assay showed that the molecular mass of the expressed protein was higher than the wild type VP4 protein, and that the modified product was corresponding to a glycosylation of VP4 protein. CONCLUSION: To modify the target gene might be an effective method to enhance the stability, antigenicity and immunogenicity of expressed protein. PMID- 12899324 TI - Comparative experimental study of Ho-Yag laser and true-cut biopsy needle in myocardial revascularization. AB - OBJECTIVE: To study the mechanism and effects of blood perfusion on acute ischemic region of myocardium through channel created by Ho-Yag laser and True cut biopsy needles with myocardial contrast echocardiography. METHODS: We partially ligated the left anterior descending coronary artery of canine hearts between the 1st and 2nd diagonal branches to produce two groups of acute myocardial ischemia models and then performed transmyocardial revascularization (TMR) on this region with Ho-Yag laser and True-cut biopsy needles. Myocardial contrast echocardiography was performed with a new generation of ultrasound contrast agent and second harmonic imaging of this region before, during ischemia and after revascularization. Pictures were taken with "R" wave trigger skill. RESULTS: Acoustic density (dB) in the ischemic region (anterior wall) with myocardial contrast echocardiography decreased obviously after the left anterior descending artery was ligated (Laser group: 5, 40 +/- 1.81, Needle group: 7.11 +/ 2.51) compared with that before (Laser group: 11.69 +/- 1.61, Needle group: 12.96 +/- 2.88, P < 0.01). dB increased remarkably after TMR by either laser or True-cut biopsy needle (Laser group: 11.02 +/- 2.01, Needle group: 10.01 +/- 4.45. P < 0.01) compared to that during ischemia and approximated to that before ischemia (P > 0.05). We found that the acoustic density of the contrast developed one picture (one cardiac cycle) ahead in the transmyocardial revascularization region than that in the lateral and other region of the left ventricle wall in the scan of both groups. CONCLUSIONS: Acute ischemic myocardium can be perfused immediately by oxygenated blood from the left ventricle through channels created with both Ho-Yag laser and True-cut biopsy needles. Evidence of blood perfusion through these channels mainly during systolic phase was detected, and myocardial contrast ultrasound using intravenous perfluorocarbon-exposed sonicated dextrose albumin was regarded as a reliable method in the study of transmyocardial revascularization. PMID- 12899325 TI - Bioactivity of bioresorbable osteosynthetic devices made of hydroxyapatite/poly DL-lactide composites: an experimental study. AB - AIM: To investigate the bioactivity of the self-designed biodegradable osteosynthetic devices made of resorbable hydroxyapatite microparticles/poly-DL lactide (HA/PDLLA) composites. METHOD: Forty-three rabbits with a transverse transcondylar osteotomy of the distal femur were fixed intramedullary by a HA/PDLLA rod, the duration of follow-up were 3, 6, 12, 24 and 36 weeks. Histological, scanning electron microscopic (SEM), energy dispersive X-ray (EDX) and biomechanical analyses were done. RESULTS: Active new bone formation and direct bone-bonding were seen at the bone-implant interface. Generous apatite crystals deposited and grew on the surface of the composites at 3 approximately 6 weeks postoperation. The interfacial shear strength increased significantly. CONCLUSION: Through the incorporating of resorbable HA microparticles, specific bone-bonding and active osteogenic capacity is introduced. This kind of bioactivity, together with other properties such as sufficient mechanical strength, enhanced biocompatibility and radiopacity, which are intrinsically unobtainable in totally resorbable polymer/polymer systems, make the HA/PDLLA composites become a desirable material for the internal fixation of cancellous bone. PMID- 12899326 TI - The influence of human single chain inteleukin-12 gene transduction on the biological behavior of hepatoma 7721 cells. AB - OBJECTIVE: To investigate the anti-tumor effects of human single chain interleukin-12 (hscIL-12). METHOD: pcDNA/hscIL-12 recombinant was transfected into human hepatic carcinoma cells (7721 cells) by lipofectin method. The 7721/hscIL-12 cells which secrete hscIL-12 stably, were obtained via G418 selection, and in vitro the influence of hscIL-12 gene transduction on the growth of tumor cells was evaluated by cell cycle analysis. In vivo, genetically engineered 7721 cells (7721/hscIL-12, 7721/pcDNA) and parental cells were implanted into BALB/c nude mice, respectively. 7721/pcDNA and 7721/hscIL-12 groups were divided into two sub-groups on day 8: one was administered with hPBL twice, 6 days at interval; the other was given equal volume of PBS. Mice were sacrificed on day 26, and spleens and tumors were taken out for histologic assay. RESULTS: hscIL-12 produced stably by 7721/hscIL-12 cells had bioactivity, and it was proved by Western blot, immunocytochemistry, and in situ hybridization. In vitro, compared with 7721 and 7721/pcDNA, the 7721/hscIL-12 grew much more slowly. FACS assay showed apparent G1 arrest of 7721/hscIL-12 cells. In animal experiment, on day 8 after inoculation, the tumors of 7721 and 7721/pcDNA group were up to 5 approximately 7 mm, while those of 7721/hscIL-12 group were 2 approximately 4 mm. When treated with hPBL, the tumor of 7721/hscIL-12 group disappeared completely. Histologically, the tumors from 7721/hscIL-12 without hPBL treatment had numerous lymphocyte infiltration, the tumor cells displayed depression looking, atrophy, focal necrosis and apoptosis, whereas the tumors of 7721 and 7721/pcDNA groups grew thrivingly. CONCLUSION: hscIL-12 transduced 7721 cells could induced significant antitumor immune response which resulted in tumor regression totally when the hPBL was inoculated, and also hscIL-12 has certain effects on mice immune system. These findings suggest that hscIL-12 and hscIL-12 gene therapy might have promising prospects in clinical application. PMID- 12899327 TI - Preliminary study of three-tone paradigm auditory P300 on aging dementia. AB - OBJECTIVE: To explore the changes of three-tone paradigm P300 and observe the improvement of the diagnostic sensitivity of P300 in aging dementia. METHODS: Thirty-four patients of aging dementia underwent traditional P300 (2t-P300) and three-tone paradigm auditory P300 (3t-P300). The results of these two methods were compared. RESULTS: Abnormal P300 was found in 18 of 34 dementia cases (52.9%) by 2t-P300, and in 26 of 30 patients who finished the 3t-P300 tests (86.7%). In 2t-P300, 9 of 24 mild cases, 7 of 8 moderate cases and 2 severe cases showed abnormal results. In 3t-P300, 20 of 24 mild cases and all 6 moderate cases showed abnormal results. Abnormal 3t-P300 was found in 13 of 16 cases which showed normal 2t-P300. Besides prolonged and bad-shaped P300, no certain response of P300 to rare non-target stimulus (r-nT) was found in 10 mild cases by 3t-P300. CONCLUSION: Three-tone paradigm auditory P300 had higher sensitivity in early diagnosis of aging dementia than traditional auditory P300. No P300 response to r nT in 3t-P300 is an important index in early diagnosis of mild dementia. PMID- 12899328 TI - The pathogenesis of experimental model of mitochondrial myopathy induced by germanium dioxide. AB - OBJECTIVE: The purpose of the study was to build up an animal model of mitochondrial myopathy in order to analyse the pathogenesis of the disease. METHODS: The skeletal muscles from Wistar rats treated with germanium dioxide for 24 weeks were analysed by histopathologic and electron-microscopic studies. A quantitative analysis was carried out in mitochondrial DNAs of these samples. The biological function of the model was determined. RESULTS: An animal model of mitochondrial myopathy was built up, in which oxygen free radicals were increased and mitochondrial DNA copies were decreased contrasted with controls. CONCLUSION: It suggested that environmental toxin may play a role in the pathogenesis of mitochondrial myopathy. The increase of oxygen free radicals is an important link causing the disease. PMID- 12899329 TI - DNA typing system for HLA-A2 alleles by polymerase chain reaction with sequence specific primers. AB - OBJECTIVE: To establish a PCR-SSP method for discriminating as many HLA-A*02 alleles, which could easily be introduced into a routine laboratory. METHODS: In this study we typed HLA-A*02 polymorphisms by a sequence-specific primer (SSP) method, which involved round 1 and round 2 PCR reactions to detect 17 HLA-A*02 alleles (they are HLA-A*0201-0217 alleles) covering exon 2 and exon 3. RESULTS: We have found that DNA sample concentration and purity were the most important variable in determining the quality of the result. For identifying correct band size, the size marker used was important. We noticed that different PCR machines performed differently. By this method, we detected 20 HLA-A*02 positive genomic DNA samples and found 4 kinds of HLA-A*02 alleles. They were HLA-A*0201, 0203, 0206 and 0210. CONCLUSION: The HLA-A*02 PCR-SSP method was proven to be a reliable and easily applicable typing method. Our results suggest that the SSP described here provides an optimal HLA-A*02 typing technique that may be useful in selecting donor-recipient pairs in bone marrow transplantation between unrelated individuals. PMID- 12899330 TI - Combined intraoperative iliac artery stents and femoro-popliteal bypass for multilevel atherosclerotic occlusive disease. AB - OBJECTIVE: To review our preliminary experience and evaluate our early results of a combined intraoperative iliac angioplasty and stenting with infrainguinal revascularization in multilevel atherosclerotic occlusive disease. METHODS: From July 1999 to April 2000, intraoperative iliac angioplasty and stenting combined with simultaneous femoro-popliteal bypass were performed on 12 lower extremities of 10 patients suffering from multilevel atherosclerotic occlusive disease. There were 8 men and 2 women, average 72 years. The indications for procedures included disabling claudication in 3 and rest pain in 7 patients. RESULTS: Eleven iliac angioplasty and stent procedures combined with simultaneous 9 femoro-popliteal by pass and 3 femoro-femoral-popliteal bypass were performed in 12 limbs of 10 patients. Angioplasty and stent placement was technically successful in all patients. One contralateral femoral-popliteal bypass was failure after femoro femoral-popliteal bypass. There were no additional instances of procedural or postoperative morbidity or mortality. Mean follow-up was 5 months (range 1 approximately 10 months). During the follow-up period, one femoro-infrapopliteal graft became occluded after 7 months and above-knee amputation was required. The cumulative primary patency rate of stented iliac arteries, femoro-femoral bypass grafts and femoro-popliteal bypass grafts were 100% (11/11), 100% (3/3) and 90.9% (10/11) in the follow-up period, respectively. The amputation rate was 8.3% (1/12). CONCLUSIONS: Intraoperative iliac artery PTA and stent placement can be safely and effectively performed simultaneously with infrainguinal revascularization for multilevel atherosclerotic occlusive disease by skilled vascular surgeon, using a portable C arm fluoroscopy inthe operating room. Furthermore, iliac artery PTA and stenting was valuable adjunct to distal bypass either to improve inflow and outflow, or to reduce the extent of traditional surgical intervention, and also, any angioplasty and stenting-related complications can be immediately corrected as well. PMID- 12899331 TI - The dynamic observation of serum level of SIL-2R and TNF in maxillary sinus cancer. PMID- 12899332 TI - Investigation of thrombomodulin and plasminogen activator inhibitor type-I in pregnancy induced hypertension and its clinical significance. AB - OBJECTIVE: To measure the circulating levels of thrombomodulin (TM) and plasminogen activator inhibitor type-I (PAI-I) in women with pregnancy induced hypertension (PIH). METHODS: Blood samples were drawn from 97 pregnant women in their third trimester, grouped as 25 mild PIH, 26 moderate PIH, 22 severe PIH and 24 normotensive healthy pregnant women for determining levels of TM by ELISA, PAI I by colorimetric assay methods, and creatinine (Cr) in serum by biochemical method. RESULTS: Circulating levels of TM, PAI-I and TM/Cr ratio increased with increasing severity of PIH. There were no significant differences between mild and normotensive pregnant women. The parameters were significantly changed in the moderate and severe PIH groups. CONCLUSION: TM and PAI-I may serve as meaningful clinical markers for the assessment of the endothelial damage in PIH, which is very important in evaluating and following the development of PIH. PMID- 12899333 TI - Relationship between human parvovirus B19 infection and aplastic anemia. AB - OBJECTIVE: To explore the relationship between human parvovirus B19 (HPV B19) infection and aplastic anemia (AA) and to investigate the role of HPV B19 in the occurrence of AA. METHODS: The presence of HPV B19 DNA was detected in the peripheral blood samples of 60 patients with AA (children 38 and adults 22) by nested polymerase chain reaction (PCR) assay, and 30 healthy persons were selected as control. RESULTS: Sixteen (26.7%) of 60 AA cases were HPV B19 DNA positive, while all the samples in the control group were negative for HPV B19 (P = 0.000914). Among the case group, the positive rates of HPV B19 DNA were 21.4% (6/28), 30.0% (3/10), 20.0% (1/5) and 35.3% (6/17) in children acute AA (AAA), children chronic AA (CAA), adults AAA and adults CAA patients respectively, which were significantly higher than that in the control group. Furthermore, there was no remarkable difference between children AA and adults AA in the 16 HPV B19 DNA positive patients; neither was there between AAA and CAA. CONCLUSION: HPV B19 infection is not only correlated with the occurrence of children AAA and CAA, but also with adults AAA and CAA, and might be an important viral cause for AA in humans. PMID- 12899334 TI - National survey on prevalence of cancer pain. AB - OBJECTIVE: To collect nationwide basic data about cancer related pain. METHODS: Sixty cancer patients in each province were randomly selected to participate in this survey. The subjects represented all stages of cancer, tumor sites, and different demographic characteristics. Two self-designed structured questionnaires including reasons, types of pain and pain management were used by patients and physicians respectively. Subjects were asked to report whether he/she had experienced any type of cancer related pain and filled out the equivalent questionnaire. The severity of pain was assessed by using "visual analogue scale". Original data input and analysis were using EPI-INFO software package. RESULTS: The result showed that 61.6% (958/1555) of patients had different types of cancer related pain. Majority of pain (85.1%) were caused by advanced cancer. The major reasons (64.4%) for poor management or impedimental factors of pain care are due to patient including over-concern on opioid analgesic addiction, reluctance to report pain or refused to use opioid analgesic until at times when pain is intolerable; 26.8% belonged to physician's reasons including fear to cause addiction on opioid and lack of knowledge about cancer pain management; 16.2% are due to lack of different kinds of opioid analgesic for use and 16.1% belonged to drug regulation. CONCLUSIONS: The results showed that majority of patients (61.6%) had different types of cancer related pain. In most of patients, cancer pain was relieved when they were treated. The major reason for under-treatment or impeded factors for effective relief of cancer pain was fear of opioid addiction by both medical professionals and patients. PMID- 12899335 TI - Introduction of the high throughput screening system. AB - In this article, we introduce the system of high throughput screening (HTS). Its role in new drug study and current development is described. The relationship between research achievements of genome study and new type screening model of new drugs is emphasized. The personal opinions of current problems about HIS study in China are raised. PMID- 12899336 TI - The roles of bcl-2 gene family in the pulmonary artery remodeling of hypoxia pulmonary hypertension in rats. AB - OBJECTIVE: To investigate the roles of apoptosis in the pulmonary artery remodeling of pulmonary hypertension secondary to hypoxia and illustrate the relative genes expression. METHODS: Thirty rats were divided into hypoxia group (10% O2, 8 h/d) and normal control group. On the 15th day of hypoxia, pulmonary artery pressure and right ventricular hypertrophy index were measured and pulmonary artery vessels were studied by light microscope. Then terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) technique was used to detect nucleosomal DNA fragmentation of apoptotic cells. In situ hybridization and RT-PCR were used to detect the expression level of bcl-2 and bax. RESULTS: The pulmonary artery pressure and right ventricular hypertrophy index of hypoxia group were increased significantly, the pulmonary artery wall of hypoxic group become incrassate than control group. Apoptotic cells can be found in lung with hypoxia or without hypoxia. Compared with control group, apoptotic index of hypoxic group decreased significantly. Through the methods of in situ hybridization and RT-PCR, we found the expression of bcl-2 increased whereas bax decreased significantly in the hypoxic group. CONCLUSION: The alternation in bcl 2 and bax expression induced by hypoxia play an important role in the pulmonary artery remodeling which is the main pathologic change of pulmonary hypertension secondary to hypoxia. PMID- 12899337 TI - Comparison of T-lymphocyte subsets and phenotypes between HIV-positive subjects and HIV-negative subjects. PMID- 12899338 TI - Importance of epidermal growth factor in pathogenesis of fetal growth restriction. PMID- 12899339 TI - Role of calcineurin in angiotensin II-induced cardiac myocyte hypertrophy of rats. AB - OBJECTIVE: The present study investigated the role of calcineurin in angiotensin II(AngII) -induced cardiac myocyte hypertrophy of rats. METHOD: The primary cardiac myocytes were cultured under the standard conditions. The calcineurin activity in AngII-treated cardiomyocytes was tested by using PNPP; protein synethsis rate was assessed by 3H-leucine incorporation; atrial natriuretic factor(ANF) mRNA level was determined by Northern blot analysis. Cell viability was estimated by lactate dehydrogenase (LDH) levels in cultured medium and by dyed cell numbers. RESULT: After stimulation of 10, 100 and 1 000 nmol/L of AngIi, calcineurin activities in the cardiomyocytes were increased by 13%, 57% (P < 0.05) and 228% (P < 0.01) respectively, compared with control group. Cyclosporin A(CsA), a specific inhibitor of calcineurin, markedly inhibited the calcineurin activity and decreased the 3H-leucine incorporation in AngII-treated cardiomyocytes in a dose-dependent manner. It was also found that CsA slightly reduced the mRNA level of ANF gene in AngII-stimulated cardiomyocytes. CONCLUSION: During AngII-induced cardiac myocyte hypertrophy, calcineurin signal pathway is activated, and inhibition of the pathway can attenuate AngII-induced cardiac myocyte hypertrophy, which suggests that the calcineurin signal pathway may play an important role in AngII-induced myocardial hypertrophy of rats. PMID- 12899340 TI - Effects of the HOE 694 on transient inward current and Na(+) - Ca(2+) exchange in guinea pig cardiomyocytes. AB - OBJECTIVE: To determine the effects of HOE 694, a new and potent Na(+)-H+ exchanger blocker, on transient inward current (It(i)) and Na(+) - Ca(2+) exchange during hypoxia-reoxygenation in guinea pig cardiomyocytes. METHODS. Cardiomyocytes were isolated from adult guinea pig ventricle. Experiment was performed in an experimental chamber that allowed the cells to be exposed to a sufficiently low O2 pressure. The cells were subjected to hypoxia and reoxygenation. The ionic currents were studied with patch clamp technique. RESULTS: In the absence of HOE 694, hypoxia-reoxygenation induced It(i) in 12 of 15 experiments; but in cardiomyocytes pretreated with HOE 694 (10 approximately 50 micro mol/L), the incidence of It(i) observed during reoxygenation was reduced to 5 of 11 experiments and 3 of 10 experiments, P < 0.05 vs control respectively. The Na(+) - Ca(2+) exchange current was unaffected by HOE 694 under normoxic condition. However, when cells were pretreated with 10 micromol/L HOE 694 for 10 min, then subjected to hypoxia condition, the Na(+) - Ca(2+) exchange current significantly inhibited. CONCLUSIONS: Blockade of the Na(+)-H+ exchange by HOE 694 could reduce Ca(2+) overload upon hypoxia-reoxygenation, and inhibition of Na(+) - H+ exchange may also indirectly decrease Na(+) - Ca(2+) exchange activity during hypoxia. PMID- 12899341 TI - Externalization and internalization of cardiac endothelin receptors during different phases of sepsis in rat. AB - OBJECTIVE: To study the redistribution of endothelin-1 (ET-1) receptors in two subcellular organelles, the sarcolemmal membrane and the light vesicle, of rat heart during the progression of sepsis. METHODS: Sepsis was induced by cecal ligation and puncture (CLP). ET1 receptor was assayed by using [125I]-ET1 binding. Marker enzyme activities, protein yield, and dry-to-wet weight ratio of cardiac membranes were measured. RESULTS: Septic rat heart exhibited two distinct phases: an initial hyperdynamic phase( 9h after CLP; early stage of sepsis) followed by a hypodynamic (18h after CLP, late stage of sepsis) phase. [125I]-ET1 binding study showed that during early stage of sepsis, the B(max) of ET1 receptors was increased by 30% in sarcolemma but decreased by 19% in light vesicles, while during late stage of sepsis, the B(max) was decreased by 24% in sarcolemma but increased by 38% in light vesicles. The total binding of sarcolemma and light vesicles was increased by 25% during early stage of sepsis but decreased by 17% during late stage of sepsis. CONCLUSIONS: These data indicated that ET1 receptors in the rat heart were externalized from light vesicles to sarcolemmal membranes during early hyperdynamic phase while internalized from surface membranes to intracellular compartment during late hypodynamic phase of sepsis. PMID- 12899342 TI - Carotid intima-media thickness and the association with cardiovascular risk factors in the elderly. AB - OBJECTIVE: To investigate the association of carotid arterial intima-media thickness (IMT) with principal cardiovascular risk factors in the elderly. METHODS: Carotid arterial IMT was measured by high resolution B mode ultrasound in 94 elderly subjects (old-aged group), and compared with subjects aged < 60 (middle-aged group). RESULTS: In comparison with the middle-aged group, the prevalence of coronary heart disease, cerebral vascular disease, hypertension and diabetes mellitus was significantly higher, and serum cholesterol and systolic blood pressure were also significantly higher in old-aged group. Although there was no obvious difference in IMT between the two groups, carotid plaque and carotid wall thickening were more frequently found in old-aged group. Age, systolic blood pressure and serum cholesterol were shown as the independent determinants for carotid IMT in the total participants, whereas no such independent relation was found in old-aged group. CONCLUSION: Age is the major risk factor for carotid atherosclerosis in the elderly. In other words, the occurrence of carotid atherosclerosis is the result of advancing age combined with the effect of multiple cardiovascular risk factors. PMID- 12899343 TI - Experimental study of the morphology of cerebral bridging vein. AB - OBJECTIVE: To investigate the morphological properties of pig cerebral bridging vein. METHODS: The morphology and fibre arrangement of 15 cerebral bridging veins obtained from 7 Danish Yorkshire landrace pigs were observed. RESULTS: There was a narrow region at the junction of the cerebral bridging veins and superior sagittal sinus termed "outflow cuff segment". The diameter and length of outflow cuff segment were much smaller and the thickness was higher than those of the cerebral bridging veins ( P < 0.01), and circumferential collagen fibres were most dense in the outflow cuff segment. The opening angle of the outflow cuff segment and the cerebral bridging veins were 115 +/- 4 degrees and 120 +/- 4 degrees ( P > 0.05). CONCLUSIONS: There were differences in fibre arrangement and morphological properties between the outflow cuff segment and the cerebral bridging vein, just like a resistance valve, the outflow cuff segment may play an important role in stabilizing cerebral venous outflow and regulating intracranial pressure. PMID- 12899344 TI - Regulating effects of vascular endothelial growth factor and ANG II on frog's pericardial stomata, mesothelium and angiogenesis. AB - OBJECTIVE: To observe the regulating effects of vascular endothelial growth factor (VEGF) and angiotensin II (ANG II) on the frog's pericardium, lymphatic stomata and angiogenesis so as to reveal their effects and mechanism on the mesothelial permeability, lymphatic stoma regulation and myocardial hypertrophy. METHODS: VEGF and ANG II were injected into the frog's peritoneal cavity so as to examine the changes of the pericardial stromata by using transmission electron microscopy, scanning electron microscopy and computerized imaging analysis. RESULTS: Scattered distributed pericardial stomata were found on the parietal pericardium of the frog with a few sinusoid mesothelial cells, whose blood supply was directly from the cardiac chambers flowing into the trabecular spaces of the myocardium (because there are no blood vessels in the myocardium of the frog). The average diameters of the pericardial stomata in VEGF and ANG II groups were 1. 50 microm and 1.79 microm respectively, which were much larger than those in the control group (0.72 microm, P < 0.01); the average distribution densities of the stomata were 8. 25/0.1 mm2 and 12.80/0.1 mm2 in VEGF and ANG II groups, which were also much higher than those in the control group (3.57/0.1 mm2, P < 0.01); the sinusoid areas in VEGF and ANG II groups were 2442.95 microm2/0.1 mm2 and 2121.79 microm2/0.1 mm2, which were larger than that in the control group (995.08 microm2/0.1 mm2 , P < 0.01); no angiogenesis was found in the frogs of the experimental groups. CONCLUSIONS: VEGF and ANG II could strongly regulate the pericardial stomata by increasing their numbers and openings with larger diameters and higher distribution density. They could also increase the sinusoid areas with the result of the higher permeability of the pericardium, which clearly indicated that VEGF and ANG II could speed up the material transfer of the pericardial cavity and play an important role in preventing myocardial interstitial edema. Yet there was no strong evidence to show the angiogenesis in the myocardium. PMID- 12899345 TI - Endogenous expression and HLA stabilization assay of Plasmodium falciparum CTL epitope minigene in human HLA-A2. 1 and HLA-B51 cells. AB - OBJECTIVE: To evaluate the Plasmodium falciparum CTL epitope vaccines in HLA class I allele specific human cell lines that have high frequency among Chinese population. METHODS: Synthesized oligonucleotides encoding for P. f. CTL epitope genes, constructed eukaryotic expression plasmids, transfected the minigenes into HLA class I allele specific human cell lines and identified endogenous expressing of the minigenes by RT-PCR and HLA stabilization assay. RESULTS: Two mini-genes encoding Plasmodium falciparum CTL epitopes were designed and cloned, respectively, into an eukaryotic expressing vector to form TR26 which was restricted to HLA-B51, SH6 which was restricted to HLA-A2. 1, and TS, which had the two aforementioned mini-genes fused in tandem. All of these CTL epitope genes were transfected and endogenously expressed in respective cell lines containing appropriate HLA molecules. The obviously increased expressions of HLA class I molecules were detected in the transfected cell lines. It was demonstrated that the two discrete Plasmodium falciparum epitope genes were effectively processed and presented, and the close proximity of the two epitope genes in one chain as in mini-gene TS did not interfere with the processing and presenting of each epitope gene in corresponding cell line. CONCLUSION: A successful expression and presentation of multiple CTL epitope mini-gene in MHC class I allele specific human cell lines were demonstrated by an in vitro assay, which could be corresponding to the vaccination of CTL vaccines in people with different MHC I molecules. This work also suggested the possibility of constructing a multiple CTL epitope Plasmodium falciparum DNA vaccine that could cover most of Chinese population. PMID- 12899346 TI - Identification of glucose transporter-1 and it's functional assay in mouse glomerular mesangial cells cultured in vitro. AB - OBJECTIVE: To evaluate the role of glucose transporter-1 (GLUT1) in the glucose uptake of glomerular mesangial cells. METHODS: Cultured C57/SJL mouse mesangial cells were used in the study. The expression of GLUT1 mRNA was detected by RT PCR. The expression of GLUT1 protein was detected by immunofluorescence and flow cytometry. The uptake of glucose and its kinetics were determined by 2-deoxy-[3H] -D-glucose uptake. RESULTS: Both GLUT1 mRNA and protein were found in mouse glomerular mesangial cells. 2-deoxy-D-glucose uptake and kinetics assay showed that this glucose transporter had high affinity for glucose and the glucose uptake specificity was further confirmed by phloretin. CONCLUSION: Functional GLUT1 did present in mouse mesangial cells cultured in vitro and it might be the predominant transporter mediated the uptake of glucose into mesangial cells. PMID- 12899347 TI - Assessing the resectability of pancreatic ductal adenocarcinoma: comparision of dual-phase helical CT arterial portography with conventional angiography. AB - OBJECTIVE: To evaluate the respective value of dual-phase helical CT arterial portography (CTAP) and conventional angiography in preoperative predicting resectability of pancreatic ductal adenocarcinoma. SUBJECTS AND METHODS: Tumor resectability was prospectively evaluated in 54 patients with pathologically proven pancreatic ductal adenocarcinoma who later underwent surgery. Both dual phase helical CT scanning and selective angiography were obtained in each patient preoperatively. For optimal enhancement of pancreas and major peripancreatic vessels, two catheters connected to an automatic injector via a Y-shaped tube were placed selective angiography, one in celiac trunk, the other in superior mesenteric artery. Then the patient underwent dual-phase helical CTAP of pancreas and liver. The criteria of irresectability for CTAP include: tumor diameter > or = 5 cm, extrapancreatic invasion, distant metastases and vascular involvement (occlusion, stenosis or semicircular encasement of superior mesenteric artery, hepatic artery, splenic artery, celiac axis; portal vein, superior mesenteric vein or splenic vein). The results of both modalities were correlated with findings from surgery or pathology. RESULTS: Thirty-eight of 54 patients had nonresectable disease. In prediction the irresectability, sensitivity, specificity, positive predictive value, negative predictive value and overall accuracy were 94.7%, 100%, 100%, 88.9%, 96.3% respectively for helical CTAP and 63.2%, 93.89%, 96.0%, 51.7%, 72. 2% respectively for selective angiography. In assessing vascular involvements, dual-phase helical CTAP was also superior to selective angiography. CONCLUSION: Dual-phase helical CTAP is superior to angiography in assessing resectability of pancreatic ductal adenocarcinoma. The combination of the two modalities may further improve overall creatic PMID- 12899348 TI - Epidermal growth factor prevents increased permeability and bacterial translocation in rats with acute pancreatitis. AB - OBJECTIVE: To evaluate the effects of epidermal growth factor (EGF) on intestinal permeability and bacterial translocation in rats with acute pancreatitis during total parenteral nutrition (TPN). METHODS: Thirty-two male Sprague-Dawley rats that underwent injection of 3.5% sodium taurocholate solution into the pancreatic duct were randomly divided into one of the following two groups: (1) received only TPN (control group) or (2) received TPN with EGF at a dose of 0.2 mg x kg( 1) x day(-1) (Egf group). On fifth day of total parenteral nutrition, samples from mesenteric lymph nodes, pancreas, liver and spleen were harvested for cultures. Water, protein and DNA content in jejunal mucosa were determined. D xylose and fluorescein isothiocyanate (FTC)-dextran were instilled into the lumen of a ligated segament of small intestine. Thirty minutes later, superior mesenteric vein D-xylose and plasma FITC-dextran concentration were measured. RESULTS: Positive cultures in liver and spleen, as well as FITC-dextran concentration in the Egf group were significantly lower than in the control group. Protein and DNA content in jejunal mucosa in the Egf group were significantly higher than in the control group. CONCLUSION: The results indicate that EGF may prevent increased intestinal permeability and bacterial translocation in rats with acute pancreatitis during TPN. PMID- 12899349 TI - Clinical characteristics and basic research development of Peutz-Jeghers syndrome. AB - OBJECTIVE: To improve clinical knowledge of Peutz-Jeghers syndrome. METHODS: Eight patients with Peutz-Jeghers syndrome from 1984 to 1998 in our hospital were retrospectively reviewed and analyzed in the present study. RESULT: The result of this analysis showed that there were 4 patients appeared with family histories of Peutz-Jeghers syndrome. All of the included patients admitted to the hospital with various complications, and eventually received surgical interventions for these complications, among which, 6 of them had intestinal obstructions mostly (5/6) due to small bowel intussusception, and 2 of them suffered with hemafecia. Post-operative recoveries were generally satisfactory with zero mortality. CONCLUSION: Peutz-Jeghers syndrome is an uncommon digestive dominant hereditary disease. The diagnosis of it with history, symptoms, signs, and proper examinations usually is not difficult Surgical interventions are necessary once complications occur. PMID- 12899350 TI - Spirochetes--the possible etiological factor of the cheilitis granulomatosa. AB - OBJECTIVE: To detect spirochetes in sections and to study the therapeutic effect of penicillin in cheilitis granulomatosa (CG) and Melkersson-Rosenthal syndrome (MRS) and the relationship between the spirochetes infection and CG and MRS. METHODS: Routine HE sections and Warthin-Starry special staining were carried out in 20 cases of CG and 6 cases of MRS. Meanwhile there were 9 cases of CG and 2 caes of MRS were treated by penicillin (12 000 000u, i.v., per day) for two courses (14 days). RESULTS: A kind of spirochete was discovered in the sections of all cases of the CG and MRS. The CG and MRS could be divided into two types histopathologically, that is granuloma type and interstitial inflammatory type (non-granuloma type), those morphological changes tallied with spirochetosis. After treatment by penicillin, the facial and labial swelling of the 11 cases of CG and MRS were abated. CONCLUSION: CG and MRS probably are infectious diseases caused by spirochetes. PMID- 12899351 TI - Study of T cell receptor gene rearrangements in patients with myasthenia gravis. PMID- 12899352 TI - Flashlamp-pumped pulsed dye laser in treatment of port-wine stains. AB - OBJECTIVE: To assess the effectiveness of the flashlamp-pumped pulsed dye laser (Photogeneca V, Synosure Corp, Boston, United States) in the treatment of port wine stains. METHODS: One hundred and ninety-four consecutive patients with port wine stains were treated with a flashlamp-pumped pulsed dye laser in Peking Union Medical College Hospital from January 1998 to August 1999. RESULTS: Of 194 patients who completed treatment, 56.2% had more than 60% fading of the lesion and only 6.7% had less than 20% fading. An average of 3.6 treatments were needed to achieve more than 60% fading. The response was better in children than in adults, although the difference was not significant. Pigmentary change (usually transient) occurred in 3.1% of patients. CONCLUSIONS: This study confirms the efficacy of the flashlamp-pumped pulsed dye laser in the treatment of port-wine stains in children and adults. PMID- 12899353 TI - Progress in DNA chip technology. AB - DNA chip technology employs light-directed in situ oligonucleotide synthesis and/or DNA microarray printing device to produce arrays of large number of probes in the tiny surface of silicon substrates, which makes it possible that the gene detection be conducted efficiently with high speed and sensitivity. The DNA chip may take important part in genome research, gene diagnoses and so on. PMID- 12899354 TI - Affective habituation: subliminal exposure to extreme stimuli decreases their extremity. AB - Following a functional perspective on evaluation, the authors hypothesized that subliminal exposure to extreme stimuli (e.g., extremely negative or positive words) would lead these stimuli to be perceived as less extreme. This process- affective habituation--was tested in 4 experiments. In Experiment 1, participants were subliminally exposed to extremely positive and extremely negative words. In a subsequent explicit-judgment task, these words were rated as less extreme than extreme words that had not been presented. In Experiment 2, these results were replicated with an implicit evaluation measure. In Experiments 3 and 4, subliminal exposure to extreme positive and negative words made the words "behave" as words that are only moderately positive or negative. Several implications are discussed. PMID- 12899355 TI - An index of self-regulation of emotion and the study of repression in social contexts that threaten or do not threaten self-concept. AB - This research tests a model of repression (M. Mendolia, 1999; M. Mendolia, J. Moore, & A. Tesser, 1996) which specifies that the interaction of individual differences in emotional responsiveness and situational threats to self-concept contributes to one's tendency to regulate emotional responsiveness. This research demonstrates that (a) individuals regulate their autonomic activity, facial muscle activity, cognitive attention, and subjective experience during isolated and repeated exposures to self-threatening negative and positive emotional events and (b) repressive behavior can be predicted by the Index of Self-Regulation of Emotion, which complements and extends conventional categorical measures of dispositional repression. This model provides a more detailed understanding of basic mechanisms in emotion by identifying how individual differences in emotionality and particular social contexts contribute to self-regulation of emotion. PMID- 12899356 TI - Right-sided human prefrontal brain activation during acquisition of conditioned fear. AB - This H2(15)O positron emission tomography (PET) study reports on relative regional cerebral blood flow (rCBF) alterations during fear conditioning in humans. In the PET scanner, subjects viewed a TV screen with either visual white noise or snake videotapes displayed alone, then with electric shocks, followed by final presentations of white noise and snakes. Autonomic nervous system responses confirmed fear conditioning only to snakes. To reveal neural activation during acquisition, while equating sensory stimulation, scans during snakes with shocks and white noise alone were contrasted against white noise with shocks and snakes alone. During acquisition, rCBF increased in the right medial frontal gyrus, supporting a role for the prefrontal cortex in fear conditioning to unmasked evolutionary fear-relevant stimuli. PMID- 12899357 TI - Latent inhibition in human affective learning. AB - This study presents the results of a new visual procedure designed to generate affective learning, namely the change in the affective rating of a previously neutral stimulus by simply pairing it with a liked or disliked stimulus. Specifically, an experiment was conducted to evaluate the effect of nonreinforced preexposures to the to-be-conditioned stimulus (a non-sense shape) on affective conditioning. This manipulation, intended to produce latent inhibition, typically results in retarded learning when the preexposed stimulus is paired with the unconditioned stimulus. The results revealed that it is possible to modify the affective value of a previously neutral non-sense shape by pairing it with a liked or disliked unconditioned stimulus, and that latent inhibition affects affective conditioning. PMID- 12899358 TI - Informational mood impact on effort-related cardiovascular response: the diagnostic value of mood counts. AB - Drawing on the mood-behavior model (G. H. E. Gendolla, 2000), 2 experiments examined moods' informational impact on effort-related cardiovascular response. After being induced into positive versus negative moods, participants performed a memory task (Experiment 1) or a letter-cancellation task (Experiment 2). Half the participants received a cue that their mood could have been manipulated. As expected, both studies found stronger reactivity of systolic blood pressure in a negative mood than in a positive mood when no cue was provided. This effect diminished in the cue conditions. Additionally, achievement corresponded to systolic blood pressure reactivity (Experiment 1), the cue manipulation had no effect on mood, and mood had a congruency effect on subjective task difficulty in the no-cue conditions (Experiment 2). PMID- 12899359 TI - The effects of adult aging and induced positive and negative mood on planning. AB - Changes in executive functions have been found in older adults and also in young adults experiencing positive or negative mood states. The current study investigated the hypothesis that older adults would show greater executive function impairment following mood induction than young adults. Ninety-six participants (half aged 19-37, half aged 53-80) completed a neutral, positive, or negative mood induction procedure, followed by the Tower of London planning task. Significant interactions were found between age and mood such that older adults showed greater planning impairment than young adults in both the positive and negative mood conditions. Emotionally salient events occurring before testing may interfere with executive function in older adults. PMID- 12899360 TI - Anxiety-related bias in the classification of emotionally ambiguous facial expressions. AB - High- and low-trait socially anxious individuals classified the emotional expressions of photographic quality continua of interpolated ("morphed") facial images that were derived from combining 6 basic prototype emotional expressions to various degrees, with the 2 adjacent emotions arranged in an emotion hexagon. When fear was 1 of the 2 component emotions, the high-trait group displayed enhanced sensitivity for fear. In a 2nd experiment where a mood manipulation was incorporated, again, the high-trait group exhibited enhanced sensitivity for fear. The low-trait group was sensitive for happiness in the control condition. The mood-manipulated group had increased sensitivity for anger expressions, and trait anxiety did not moderate these effects. Interpretations of the results related to the classification of fearful expressions are discussed. PMID- 12899361 TI - The eyebrow frown: a salient social signal. AB - Seven experiments investigated the finding that threatening schematic faces are detected more quickly than nonthreatening faces. Threatening faces with v-shaped eyebrows (angry and scheming expressions) were detected more quickly than nonthreatening faces with inverted v-shaped eyebrows (happy and sad expressions). In contrast to the hypothesis that these effects were due to perceptual features unrelated to the face, no advantage was found for v-shaped eyebrows presented in a nonfacelike object. Furthermore, the addition of internal facial features (the eyes, or the nose and mouth) was necessary to produce the detection advantage for faces with v-shaped eyebrows. Overall, the results are interpreted as showing that the v-shaped eyebrow configuration affords easy detection, but only when other internal facial features are present. PMID- 12899362 TI - Facial expressions of emotion: does the prototype represent central tendency, frequency of instantiation, or an ideal? PMID- 12899363 TI - Implicit anxiety measure predicts cardiovascular reactivity to an evaluated speaking task. AB - Explicit personality tests assess introspectively accessible self-descriptions. By contrast, implicit personality tests assess introspectively inaccessible processes that operate outside of awareness. Despite their inaccessibility, implicit processes are presumed to influence a variety of current responses. This study tested the hypothesis that an implicit anxiety test should predict cardiovascular reactivity during a speech stressor task. In all, 97 participants completed a measure of attention allocation toward threat (implicit test) and an anxiety questionnaire (explicit test) 1 week before giving an evaluated speech. Whereas the explicit test showed modest relations within only 1 measure of cardiovascular reactivity, the implicit test predicted heart rate and blood pressure reactivity during preparation and delivery of the speech. These findings encourage the broader use of implicit measures to assess cardiovascular responses to threat. PMID- 12899364 TI - Positive mood and executive function: evidence from stroop and fluency tasks. AB - Contrasting predictions have been made about the effects of positive mood states on the performance of frontal lobe tests that tap executive functions such as inhibition, switching, and strategy use. It has been argued that positive mood is likely to improve some cognitive processes, particularly those dependent on the frontal cortex and anterior cingulate of the brain. However, there is some evidence that happy mood may impair executive functioning. The current experiments investigated the effects of positive mood on Stoop and fluency tests, which are frequently used to assess executive function. Positive mood impaired performance on a switching condition of the Stroop test, but improved performance on a creative uses test of fluency. The effect of positive mood on an executive task may therefore depend on whether a task is inherently motivating or is impaired by diffuse semantic activation. PMID- 12899365 TI - Neural systems for recognition of emotional prosody: a 3-D lesion study. AB - Which brain regions are associated with recognition of emotional prosody? Are these distinct from those for recognition of facial expression? These issues were investigated by mapping the overlaps of co-registered lesions from 66 brain damaged participants as a function of their performance in rating basic emotions. It was found that recognizing emotions from prosody draws on the right frontoparietal operculum, the bilateral frontal pole, and the left frontal operculum. Recognizing emotions from prosody and facial expressions draws on the right frontoparietal cortex, which may be important in reconstructing aspects of the emotion signaled by the stimulus. Furthermore, there were regions in the left and right temporal lobes that contributed disproportionately to recognition of emotion from faces or prosody, respectively. PMID- 12899366 TI - Duchenne smile, emotional experience, and autonomic reactivity: a test of the facial feedback hypothesis. AB - This study examined the modulatory function of Duchenne and non-Duchenne smiles on subjective and autonomic components of emotion. Participants were asked to hold a pencil in their mouth to either facilitate or inhibit smiles and were not instructed to contract specific muscles. Five conditions--namely lips pressing, low-level non-Duchenne smiling, high-level non-Duchenne smiling, Duchenne smiling, and control--were produced while participants watched videoclips that were evocative of positive or negative affect. Participants who displayed Duchenne smiles reported more positive experience when pleasant scenes and humorous cartoons were presented. Furthermore, they tended to exhibit different patterns of autonomic arousal when viewing positive scenes. These results support thefacial feedback hypothesis and suggest that facial feedback has more powerful effects when facial configurations represent valid analogs of basic emotional expressions. PMID- 12899367 TI - Cross-cultural patterns in emotion recognition: highlighting design and analytical techniques. AB - This article highlights a range of design and analytical tools for studying the cross-cultural communication of emotion using forced-choice experimental designs. American, Indian, and Japanese participants judged facial expressions from all 3 cultures. A factorial experimental design is used, balanced n x n across cultures, to separate "absolute" cultural differences from "relational" effects characterizing the relationship between the emotion expressor and perceiver. Use of a response bias correction is illustrated for the tendency to endorse particular multiple-choice categories more often than others. Treating response bias also as an opportunity to gain insight into attributional style, the authors examined similarities and differences in response patterns across cultural groups. Finally, the authors examined patterns in the errors or confusions that participants make during emotion recognition and documented strong similarity across cultures. PMID- 12899368 TI - Fundamental dimensions of subjective state in performance settings: task engagement, distress, and worry. AB - Subjective state constructs are defined within the traditional domains of affect, motivation, and cognition. Currently, there is no overarching state model that interrelates constructs within the different domains. This article reports 3 studies that provide converging evidence for 3 fundamental state dimensions labeled task engagement, distress, and worry that integrate constructs across the traditional domains. Study 1 differentiated the state dimensions by factor analysis of the scales of the Dundee Stress State Questionnaire (G. Matthews et al., 1999). Study 2 showed differential state response to performance of tasks making different cognitive demands. Study 3 showed that states are correlated with differing patterns of appraisal and coping. The 3 stress state dimensions provide a general descriptive framework consistent with transactional accounts of stress and performance. PMID- 12899369 TI - Culture and alexithymia: mean levels, correlates, and the role of parental socialization of emotions. AB - Two studies examined the relationship between culture and alexithymia. In Study 1, mean levels and correlates of alexithymia were examined in 3 cultures: European American (EA), Asian American (AA), and Malaysian college students. Both Asian groups had higher alexithymia levels than the EA group. Somatization was more strongly associated with alexithymia in the Asian groups than in the EA group. Mood and life satisfaction were associated with alexithymia in similar ways across groups. In Study 2, the relations among culture, gender, retrospective reports of parental socialization of emotions, and alexithymia were examined among EA and AA college students. Cultural and gender differences were found in alexithymia and emotion socialization levels. Most important, parental emotion socialization mediated the relations among culture, gender, and alexithymia. PMID- 12899370 TI - Alexithymia, memory of emotion, emotional awareness, and perfectionism. AB - The Toronto Alexithymia Scale (TAS-20; R. M. Bagby, J. D. A. Parker, & G. J. Taylor, 1994) is a self-assessment instrument designed to measure deficits in meta-emotional functioning (e.g., difficulties in identifying and describing emotions). Four studies were carried out to examine the association between the TAS-20 and (a) performance measures of meta-emotional functioning (memory of emotion and emotional awareness; Studies 1-2) and (b) measures of perfectionistic standards that may possibly be involved in the self-assessment of abilities difficulties (Studies 3-4). The TAS-20 failed to correlate in the predicted direction with the performance measures but showed sizable correlations with measures of perfectionism. Moreover, perfectionism was found to predict TAS-20 scores independent of depression, anxiety, and somatic complaints. The results are discussed in terms of the TAS-20 measuring primarily certain aspects of meta emotional self-efficacy. PMID- 12899371 TI - Emotional expression and physiology in European Americans and Hmong Americans. AB - Ethnographic and clinical observations suggest that Asians are less expressive than European Americans. To examine whether this difference emerged in online emotional responding, 50 Hmong Americans (HAs) and 48 European Americans (EAs) were asked to relive past episodes of intense happiness, pride, love, anger, disgust, and sadness. Facial behavior and physiological reactivity were measured. For most emotions, more cultural similarities than differences were found. There were some exceptions: During happiness, fewer HAs than EAs showed non-Duchenne smiles (i.e., "social" smiles), despite similarities in reported emotional experience and physiological reactivity. Within-group differences between "less Hmong" and "more Hmong" HAs were also found. Implications of these findings for our understanding of culture-emotion relations are discussed. PMID- 12899372 TI - Facial affect recognition in criminal psychopaths. AB - Prior studies provide consistent evidence of deficits for psychopaths in processing verbal emotional material but are inconsistent regarding nonverbal emotional material. To examine whether psychopaths exhibit general versus specific deficits in nonverbal emotional processing, 34 psychopaths and 33 nonpsychopaths identified with Hare's (R. D. Hare, 1991) Psychopathy Checklist- Revised were asked to complete a facial affect recognition test. Slides of prototypic facial expressions were presented. Three hypotheses regarding hemispheric lateralization anomalies in psychopaths were also tested (right hemisphere dysfunction, reduced lateralization, and reversed lateralization). Psychopaths were less accurate than nonpsychopaths at classifying facial affect under conditions promoting reliance on right-hemisphere resources and displayed a specific deficit in classifying disgust. These findings demonstrate that psychopaths exhibit specific deficits in nonverbal emotional processing. PMID- 12899373 TI - Experiencing activation: energetic arousal and tense arousal are not mixtures of valence and activation. AB - R. E. Thayer (1989) proposed 2 types of activation: energetic arousal (awake tired) and tense arousal (tense-calm). This view has been challenged by claims that energetic arousal and tense arousal are mixtures of valence and a single activation dimension. The authors present a direct test of this hypothesis by computing the correlation between the residuals of energetic arousal and tense arousal after removing the shared variance with valence. Whereas the valence activation hypothesis predicts a strong positive correlation between the 2 residuals, the authors found that it was not significantly different from 0. This finding reaffirms the view of energetic arousal and tense arousal as 2 distinct types of activation. PMID- 12899374 TI - Study on GMA-DNA adducts. AB - OBJECTIVE: DNA modification fixed as mutations in the cells may be an essential factor in the initiation step of chemical carcinogenesis. In order to explore the mechanism of gene mutation and cell transformation induced by glycidyl methacrylate (GMA), the current test studied the characteristics of GMA-DNA adducts formation in vitro. METHODS: In vitro test, dAMP, dCMP, dGMP, dTMP and calf thymus DNA were allowed to react with GMA (Glycidyl Methacrylate). After the reaction, the mixtures were detected by UV and subjected to reversed-phase HPLC on ultrasphere ODS reversed-phase column, the reaction products were eluted with a linear gradients of methanol (solvent A) and 10 mmol/L ammonium formate, pH 5.0 (solvent B). The synthesized adducts were then characterized by UV spectroscopy in acid (pH 1.0), neutral (pH 7.2), alkaline (pH 11.0) and by mass spectroscopy. RESULTS: The results showed that GMA could bind with dAMP, dCMP, dGMP and calf thymus DNA by covalent bond, and the binding sites were specific (N6 of adenine, N3 of cytosine). Meanwhile, a main GMA-DNA adduct in the reaction of GMA with calf thymus DNA was confirmed as N3-methacrylate-2-hydroxypropyl-dCMP. CONCLUSIONS: GMA can react with DNA and/or deoxynucleotide monophosphate and generate some adducts such as N6-methacrylate-2-hydroxypropyl-dAMP and N3 methacrylate-2-hydroxypropyl-dCMP, ets. Formation of GMA-DNA adducts is an important molecular event in gene mutation and cell transformation induced by GMA. PMID- 12899375 TI - Change of tumor necrosis factor-alpha levels in patients with Binswanger's disease. PMID- 12899376 TI - Detection of chromosome aberrations in twelve primary gastric cancers by direct chromosome analysis and FISH. AB - Direct chromosome analysis and FISH were performed on twelve primary gastric carcinomas. Two of them had simple chromosome changes: 48, XX, +8, +20, and 49, XY, +2, +8, +9, and the others had complicated chromosome changes, which includes much more numerical and structural chromosome aberrations. Frequent structural changes in the complicated types involved chromosome 7, 3, 1, 5 and 12 etc. The del 7q was noted in eight cases. The del (3p) and del (1p) were noted in six and five cases, respectively. The results provide some important clues for isolation of the genes related to gastric cancer. PMID- 12899377 TI - Intracellular expression of multimerized antisense TAR-Core RNAs inhibit the replication of human immunodeficiency virus type 1 in human CD4+ T lymphocytes. AB - Gene therapy is one of several approaches that are being tested in the search for an effective anti-HIV treatment. In this strategy, a "resistant" gene would be introduced into target cells, rendering them resistance to the infection of HIV. The HIV-1 Tat protein transactivate HIV-1 gene expression at the transcriptional level by interacting with its response element (TAR) in the long terminal repeat (LTR). Previously, we have shown that antisense polyTAR-Core RNAs can inhibit the transactivation of HIV-1 Tat protein in transiently transfected Jurkat cells. To determine whether this antisense polyTAR-Core RNAs could inhibit HIV-1 replication in CD4+ T cells, we transfected the antisense polyTAR-Core gene to MT4 cells and challenged them with HIV-1 SF33 strain. Levels of HIV-1 p24gag antigen were reduced more than 4-fold in cultures of the transduced MT4/LR cells infected with HIV-1 SF33 strain. In contrast, cultures of nontransduced MT4 cells and control LX vector transduced MT4/LX cells infected with the same viruses had high levels of HIV-1 p24gag. Our work showed that antisense polyTAR-Core RNAs were able to inhibit HIV-1 replication in CD4+ T cells, and could be used as resistance gene in further studying for gene therapy against HIV-1. PMID- 12899378 TI - Expression of cathepsin D and its prognostic significance in NSCLC patients. PMID- 12899379 TI - Defense mechanisms of urinary bladder: studies on antimicrobial polypeptides from bladder mucosa. AB - The acid-soluble extract of the bladder mucosal surface was obtained by washing out the bladder with dilute acetic acid in the presence of protease inhibitors. The wash-out materials from rats, rabbits, pigs, and humans manifested strong bactericidal activity against E. coli in vitro. The ultrafiltrate of the human material, which contained two major peptides with apparent molecular masses of 6.7 kD and 8.5 kD, respectively, showed potent bactericidal activity against E. coli, Pseudomonas aeruginosa, Staphylococcus aureus, and Streptococcus sanguis. Three antibacterial polypeptides (PiBPs) were purified from the porcine material. The molecular masses of PiBP-5, PiBP-11 and PiBP-25 were 5773.3 Da, 11127.8 Da and 25073 Da, respectively. PiBP-5 was unusually rich in glycine, serine and threonine residues (20.0, 16.3 and 10.4 mol%, respectively), and N-terminal amino acid sequencing revealed that PiBP-5 was homologous (83.3% identity in an 18 residue overlay) to the "tail" of human cytokeratin-7. Although the amino acid compositions of PiBP-11 and PiBP-25 were established, both had blocked N-termini and primary sequence data were not obtained. These results provided evidence indicating that the presence of peptides in the bladder mucosa could enable it to kill adherent bacteria. PMID- 12899380 TI - Role of bivalent cations and choline in ATP-induced apoptosis of human lymphocytes with P2Z receptors. AB - The role of bivalent cations and choline in ATP-induced apoptosis via P2Z purinoceptor was investigated in human leukemic lymphocytes. In vitro exposure of leukemic lymphocytes with P2Z receptors to 1 mmol/L ATP or 0.1 mmol/L benzoylbenzoic ATP (BzATP) for 8 h in the presence of choline, 1 mmol/L Mg2+ or other bivalent cations, and ATP-induced DNA breaks, associated with apoptosis were quantified by TdT assay. We observed that (1) Extracellular Mg2+ or Ca2+ stimulated ATP-induced DNA fragmentation in a dose-dependent manner, and the compatible evidence was provided by the inhibition of ATP-induced DNA fragmentation in the present of EGTA or EDTA; (2) ATP-induced DNA fragmentation was completely inhibited by 1 mmol/L Zn2+; (3) ATP-induced DNA breaks were not affected by Ba2+, Sr2+, Co2+ when they were substituted for extracellular Mg2+ or Ca2+; (4) Choline, an inhibitor of phospholipase D (PLD) stimulated by ATP through P2Z receptor in human lymphocytes, was also a partial inhibitor of ATP induced DNA fragmentation, and the results were confirmed by flow cytometric analysis (FCA); (5) ATP-induced DNA fragmentation was completely obliterated when the temperature was lower than 10 degrees C. These results suggest that the endonuclease and PLD may be involved in ATP-induced apoptosis in human lymphocytes via P2Z receptor. PMID- 12899381 TI - Predominant expression of human Agamma--in contrast with beta-globin gene in MEL cells transfected with the construct muLCRAgamma psibeta deltabeta. AB - A cosmid construct muLCRAgamma psibeta deltabeta were induced into mouse erythroleukemia cell lines 585 that expresses murine adult globin only and MEL GM 979 that expresses both murine embryonic and adult globins. Similar patterns of human globin gene expression were displayed in the two MEL cell lines transfected with the construct. Inducible expression of the Agamma- and beta-gene was observed during induced cell differentiation. However, the expression level of the Agamma-globin gene is much higher than that of the beta-globin gene in either uninduced or induced MEL transformants. No gamma to beta switching happened in the stable MEL transformants following a continuous culture. The much more effective enhance of the muLCR on the Agamma-globin gene than that on the beta globin gene is resulted probably from the fact that the distance between the LCR and the beta-globin gene is much longer than that between the LCR and the Agamma globin gene in the construct, in comparison with other constructs containing HS2 or muLCR linked to both of gamma- and beta-globin genes in different order. Two suggestions can be derived from these results: 1) A competition between the gamma and beta-globin gene for interaction with the LCR may indeed present, but only an enough long distance difference between the LCR to the gamma- and to the beta gene can effectively influence the competition; 2) Unlike transgenic mice, MEL cells are incapable of reconstructing the regulatory information involved in developmental control when it is provided by a fragment of the beta-globin gene cluster with limited length. PMID- 12899382 TI - The effects of S-3-1 on lipid peroxidation and scavenging free radicals in vitro. AB - S-3-1 is a simplified synthetic analogue of the active principle of Salvia miltiorrhioza. Electron spin resonance spectrometry using 5,5'-dimethyl-1 pyrroline-N-oxide as a scavenger of free radicals indicated that 200 microg/ml of S-3-1 scavenged 1,1-diphenyl-2-picrylhydrazyl radicals completely. 25 microg/ml of this compound quenched 100% of superoxide anion and a concentration of 250 microg/ml of S-3-1 quenched 63% of hydroxyl radicals. It was also shown that 3.56 mg/ml of S-3-1 could inhibit lipid peroxidation in microsome fraction from rat liver induced by FeSO4 and cysteine. These results show that S-3-1 is an effective antioxidant by scavenging free radicals. PMID- 12899383 TI - Role of endogenous carbon monoxide in neointimal formation induced by balloon injury in rat aorta. AB - OBJECTIVE: The present study investigated the role of endogenous carbon monoxide (CO) in the pathogenesis of neointimal formation induced by balloon injury in rat. METHOD: Endothelial denudation of the left common carotid artery of rat was carried out by three passages of a Fogarty 2F balloon catheter. DNA, collagen and elastin contents of each intima-media were estimated; and heme oxygenase (HO) activity and CO production in vascular smooth muscle cell (VSMC) were measured after administration of HO inhibitor. RESULT: Our data showed that neointima occurred in the rat on day 7 and day 21 after balloon injury, and at the same time HO activity and CO production in VSMC were markedly increased. Administration of HO inhibitor, zinc deuteroporphyrin 2,4-bisglycol (ZnDPBG), could effectively inhibit HO activity and CO production, significantly enhance neointimal formation (aortic intima/media ratio were 21.4+/-1.8% vs 17.6+/-2.0%, P<0.05 on day 7; and 30.5+/-2.4% vs 23.0+/-2.2%, P<0.01 on day 21, respectively, compared with balloon alone group). CONCLUSION: We concluded that 1) inhibition of CO production may enhance neointimal formation induced by endothelial denudation, implying endogenous CO play an protective role in response to vascular injury, and 2) induction of HO activity may be applied clinically for preventing restenosis after angioplasty. PMID- 12899384 TI - The association of Ala54Thr variant of intestinal fatty acid binding protein gene with general and regional adipose tissue depots. AB - OBJECTIVE: To ascertain the relationship between the Ala54Thr variation of FABP2 gene and general as well as regional adipose tissue depots. SUBJECTS: 165 subjects, in which 86 were subjects with normal glucose tolerance (NGT) [age 54.45+/-9.80, male/female 1.05, body mass index (BMI) 26.48+/-4.01] and 79 were subjects with non-insulin-dependent diabetes mellitus (NIDDM) (age 55.86+/-10.00, male/female 1.08, BMI 26.75+/-3.30). DESIGN AND MEASUREMENTS: An association study of FABP2-Ala54Thr variation detected by PCR/HhaI digestion with general and regional adipose tissue depots determined by BMI and magnetic resonance imaging [abdominal subcutaneous and visceral adipose tissue area (SA and VA) and femoral subcutaneous adipose tissue area (FA)]. RESULTS: The geneotype and allele frequencies of FABP2-Ala54Thr variation in Chinese were quite close to the frequencies in American Caucasians and Pima Indians reported in the literature. Significant difference in genotype frequency distribution was observed between FA subgroups comparisons (FA > or = 75 cm2 versus FA < 75 cm2) in NIDDM subjects (X2 = 11.460, P = 0.003), with significantly increased in Thr54-carrier [Thr54 (+)] genotype frequency and Thr54 allele frequency in NIDDM subject with FA < 75 cm2 (odd ratio for genotype was 4.62, X2 = 10.112, P = 0.001; and for allele = 2.36, X2 = 5.379, P = 0.020). The FA in NIDDM-Thr54 (+) subgroup was significantly lower than that in subjects with NIDDM-Thr54 (-) subgroup (61.19+/-21.51 cm2 versus 75.36+/-31.70 cm2, P = 0.021). Stepwise regression analysis revealed that FABP2-Thr54 genotype variation was an independent factor contributing to the variation of FA in NIDDM (P = 0.003). CONCLUSION: FABP2 is associated with regional adipose tissue depot. The decreased femoral subcutaneous adipose tissue depot in NIDDM subjects is related to FABP2-Thr54 variant. PMID- 12899385 TI - Kinetic changes of antibody to M. leprae and its relation with clinical disease. AB - A sero-epidemiological survey on 1,833 healthy residents was carried out in 6 villages of a leprosy high-endemic area in Wenshan and Guangnan counties, Yunnan Province. The part of the residents with initially antibody-positive as well as the part of residents with initially antibody-negative have been followed up for 3 consecutive years by serology and clinical examination for studying kinetic changes of antibody to M. leprae and its relation with clinical disease. The results showed that the rates of subclinical infection of leprosy in a high endemic area are different from village to village, and the risk of developing clinical disease does not associate with subclinical infection rate. It correlates with the number of cured accumulative leprosy cases and active cases within the village. The authors consider that in leprosy high-endemic villages, especially those cropped up new multi-bacillary leprosy cases frequently in recent years, it may be helpful to use serology to detect early leprosy cases. PMID- 12899386 TI - Role of outer membrane proteins in imipenem diffusion in Pseudomonas aeruginosa. AB - The present study identified the properties of porins in the outer membrane in Pseudomonas aeruginosa, and showed the role of outer membrane in determining imipenem diffusion in Pseudomonas aeruginosa. The molecular weight of the major outer membrane protein was analyzed by SDS-PAGE. The purification of the porins in Pseudomonas aeruginosa was achieved by DEAE ion-exchange HPLC. The purified outer membrane proteins were reconstituted with phosphatidylcholine and dicetylphosphate into membrane vesicles, and were tested by the liposomes swelling method for the diffusion of imipenem. The permeability assay showed that OprC (70 kD), OprD2 (46 kD), and OprE (43 kD) were the channel-forming proteins. But only OprD2 was thought to be the likely route of imipenem diffusion. PMID- 12899387 TI - Experimental study of nerve graft with vascular implantation. AB - OBJECTIVE: To study the performance of free nerve graft with vascular implantation. MATERIAL AND METHOD: Three groups of rats were used. A 10 mm nerve defect was made in the sciatic nerve. It was repaired respectively with vascularised nerve graft (VNG), free nerve graft with vascular implantation (NGV) or free nerve graft (NG). Before 30 days the revascularisation of the nerve graft was assessed; at 2 to 3 month regeneration of nerve was evaluated. Histochemical staining, vascular morphology and electrophysiological examination were undertaken. RESULT AND CONCLUSION: The number of regenerated blood vessels in VNG and NGV was significantly increased than that in NG before 30 days. The ratio of regenerated nerve fiber area and conduction velocity in the first two groups were much better than those in NG at 2 to 3 months. PMID- 12899388 TI - An experimental study of the tumour necrosis factor levels in aqueous humor after traumatic cataract and intraocular lens implantation. AB - PURPOSE: This paper studies the tumour necrosis factor (TNF) levels in aqueous humor after traumatic cataract extraction and posterior chamber (PC) intraocular lens (IOL) implantation in rabbits, and discusses the effect of TNF on postoperative anterior ocular inflammation. METHODS: Twenty-seven pigmented rabbits were divided into three groups: for the first group, the IOL were placed in the capsular bag after traumatic cataract extraction; for the second, the Extracapsular cataract extraction without IOL implantation; and for the third, the control group without surgical intervention. On the 1st, 3rd, 7th and 14th day postoperatively, aqueous humor samples were obtained. A modified double antibodies indirect sandwich ELISA was used to detected for the presence of TNF. The data were studied by means of analysis of variance in SAS software. RESULT: The TNF level was increased in aqueous humor and reached its maximum on the 1st postoperative days after traumatic cataract extraction and PC IOLs implantation, and the TNF levels are significantly higher (P<0.05) on the 1st, 3rd, 7th and 14th day postoperatively in traumatic cataract extraction and PC IOL implanted group than that in the non-surgical-intervention group and extracapsular cataract extraction group. CONCLUSIONS: The increase of TNF levels had a close relationship with presence of the IOL itself which induces the secretion of TNF. This suggested that TNF as the principal mediators of immunological and inflammatory responses, so that may play critical role in anterior ocular inflammative response after traumatic cataract extraction and IOL implantation. PMID- 12899389 TI - Involvement of the Ca(2+)-protein kinase C and adenyilate cyclace signal pathways in the activation of thymocytes in response to whole-body irradiation with low dose X-rays. AB - OBJECTIVE: To study the molecular mechanism of the stimulatory effect of low dose radiation (LDR) on T cell activation. METHODS: Thymocytes from Kunming mice exposed to whole-body irradiation (WBI) with different doses of X-rays were analyzed for the changes in signal molecules of the phospholipase C phosphatidylinositol biphosphate (PLC-IP2) and G protein-adenylate cyclase (AC) pathways. RESULTS: It was found that [Ca2+]i increased in response to doses within 0. 2 Gy which was most marked after 0. 075 Gy and the increase was accentuated in the presence of Con A. The changes in CD3 and calcineurin (CN) expression of the thymocytes followed the same pattern as the alterations in [Ca2+]i after LDR. The expression of alpha, beta1 and beta2 isoforms of protein kinase C (PKC) was all up-regulated after 0.075 Gy with the increase in PKC-beta1 expression being most marked. The cAMP/cGMP ratio and PKA activity of the thymocytes was lowered after low dose radiation and increased after doses above 0. 5 Gy in a dose-dependent manner, thus giving rise to J-shaped dose-response curves. The Ca antagonist TMB-8 and cAMP stimulant cholera toxin suppressed the augmented thymocyte proliferation induced by LDR. CONCLUSION: Data presented in the present paper suggest that activation of the PLC-PIP2 signal pathway and suppression of the AC-cAMP signal pathway are involved in the stimulation of the thymocytes following WBI with low dose X-rays. PMID- 12899390 TI - Responsibility of insulin to the change of glucose level in newborn small for gestational age infants. PMID- 12899391 TI - Effects of XW630 on bone formation in ovariectomized rats and in human osteoblast like cells. AB - OBJECTIVE: To study the effects of XW630 on bone formation in overiectomized (OVX) rats and in human osteoblast-like cell line TE85. METHOD: Bone histomorphometric analysis was performed with undecalcified bone sections and tetracycline intraperitoneally labeling. RESULTS: Compared with that of OVX rats, the static data of trabecular bone volume (TBV)/total tissue volume (TTV), TBV/sponge bone volume (SBV) and mean trabecular plate density (MTPD) were enhanced while mean trabecular plate spacing (MTPS) decreased after treated with XW630 for 13w. The dynamic data of single-labeled surface [Sfract(s)], double labeled surface [Sfract(d)], Sfract(d+1/2s), trabecular osteoid surface (TOS) and bone formation rate in tissue level (Svf) were increased and osteoid maturation period (OMP) shortened in XW630 group. In osteoblast-like cells, both 3H thymidine incorporation and cell count increased after treated with XW630 for 48. Treated with XW630 for 12 approximately 18h, inducible nitric oxide synthase (iNOS) activity and cGMP content increased in time-dependent manners. CONCLUSIONS: XW630 enhanced bone activation frequency and increased trabecular connectivity, stability, and strength. The cellular mechanism related to effects of XW630 on bone formation in ovariectomized rats. PMID- 12899392 TI - Protection against leptospirosis by immunization with plasmid DNA encoding 33 kDa endoflagellin of L. interrogans serovar lai. AB - OBJECTIVE: To evaluate how the efficacy of DNA inocutation affects the ability to raise protective immunity against Leptospira. METHODS: A pair of oligonucleotide primers were designed to amplify the endoflagellar gene of L. interrogans sensu stricto serovar lai. An approximately 840bp fragment was generated with PCR and inserted into VR1012, a plasmid DNA expression vector, after the fragment and VR1012 were digested respectively with EcoRV and Sal I. A recombinant plasmid designated as VR1012+flaB2 was obtained. The vector, VR1012 consits of a pUC18 backbone with the cytomegalovirus (CMV) IE1 enhancer, promoter, and intron A, transcription regulatory elements and the BGH polyadenylation sequences driving the expressing of leptospiral endoflagellar gene of L. interrogans sensu stricto serovar lui. Plasmid encoding leptospiral endoflagellin gene was injected into quadriceps of NZW rabbits. RESULTS: This resulted in the generation of specific leptospiral antibody with high ELISA titer (1:32768) in the rabbits. Immuno/protection was performed in guinea pigs without adjuvant. The group "VR1012 + flaB2" showed higher survival rate (90%, 9/10 animals), compared with the group "VR1012 lack flaB2" and the group "normal saline". CONCLUSION: The technique of DNA vaccine has potential advantages over certain other vaccine preparation technologies. However whether DNA vaccine will be useful for vaccine development remains to be tested. PMID- 12899393 TI - The transmembrane signal transduction in HEp-2 cells induced by bacterial adherence. AB - In order to understand the role of transmembrane signal transduction of host cells in the early steps of infection, the adherence of E. coli to HEp-2 cells and the change of activity of phospholipase C-gamma (PLC-gamma) induced by the adherence were investigated. The adherence of enteropathogenic E. coli (EPEC), strain E. 7, induced a significant increase of inositol-triphosphat (IP-3) level in HEp-2 cells. The adherence of the bacteria and the increase of IP-3 was kinetically correlated. Whereas the increase of IP3 level induced by the adherence of the control strain EPEC (H511), a non-piliated strain, was much meager than that by E7, a piliated strain. The results highlighted an important role of transmembrane signals like 1P-3 in the pathogenesis of EPEC. PMID- 12899394 TI - Polymorphism of angiotensin-converting enzyme gene and genetic susceptibility to asthma with familial aggregation. AB - OBJECTIVE: Angiotensin-converting enzyme (ACE) plays a key role in the metabolism of angiotensin II (AT II) and inactivation of bradykinins and tachykinins, which are potent bronchialconstrictors and mediators of inflammation asthma, and ACE is heavily expressed in the lungs. An insertion-deletion (D/I) polymorphism of ACE gene has been shown to be associated with levels of ACE. We investigate whether the polymorphism of ACE gene is associated with asthma and bronchial responsiveness. METHODS: A case-control study was carried out in 50 asthmatics, 7 families with at least 2 asthmatic individuals, and 50 healthy subjects. The insertion/deletion (I/D) polymorphism of ACE gene was amplified by polymerase chain reaction (PCR). Methacholine brocho-provocation and pulmonary function tests were performed in all asthmatics. RESULTS. There was an higher gene frequency of DD genotype of ACE gene in asthmatic subjects and families individuals compared with healthy subjects (46%, 53% vs 16%, P<0.05; odd ratio 4.98). An- higher prevalence of DD genotype of ACE was in patients with bronchial hyperresposiveness (BHR) (67% vs 33%, P<0.05; odd ratio 3.8). Accordingly, the mean values of FEV1% and FEV1/FVC were higher in asthmatics carrying non-DD alleles than patients with DD genotype (73.78% vs 56.56%, P<0.05, 79.19% vs 69.29%, P<0.05, respectively). CONCLUSION: These results suggested that DD allele of ACE genotype was significantly involved in genetic susceptibility to asthma. DD genotype of ACE might be a risk factor for the degree of airway obstruction, it could also be implicated in pathogenesis of bronchial hyperresponsiveness. PMID- 12899395 TI - Variation of myelin basic protein and its antibody in serum in senile dementia patients. PMID- 12899396 TI - Dobutamine magnetic resonance imaging predicts contractile reserve of chronically dysfunctional myocardium: comparison with was fluorine-18 fluorodeoxyglucose positron emission tomography. AB - OBJECTIVE: This study sought to investigate whether low-dose dobutamine-MRI can detect residual myocardial viability in patients with chronic myocardial infarction and, left ventricular dysfunction. METHODS: Eleven patients with chronic myocardial infarction and left ventricular dysfunction were employed for identification of viable myocardium by cine-MRI during dobutamine infusion. All patients underwent coronary angiography and left ventriculography, 18FDG-PET, MRI at rest and stress. The systolic wall thickening measured at rest and during stress was compared with the results of 18FDG-PET, respectively. RESULTS: A significant difference of either dobutamine-induced systolic wall thickening (SWthstress) or dobutamine-induced contractile reserve (deltaSWth = SWthstress - SWthrest) was present between viable and scar regions (1.0 +/- 0.3 versus -0.3 +/ 0.1, P < 0.01; 1.0 +/- 0.3 versus -0.2 +/- 0.2, P < 0.01). CONCLUSIONS: obutamine-induced contractile reserve can be predicted in the regions of akinesia or dyskinesia at rest when systolic wall thickening was > or = 1.0 mm during dobutamine stimulation. PMID- 12899397 TI - Excitatory connections between spinal motoneurons in the adult rat. AB - OBJECTIVES: Dendro-dendritic and dendro-somatic projections are common between spinal motoneurons. We attempted to clarify whether there are functional connections through these projections. METHODS: Motoneurons were antidromically stimulated by the muscle nerve and recorded intracellularly to examine the direct interaction between them after the related dorsal roots had been cut. RESULTS: Excitatory connections, demonstrated by depolarizing potentials in response to muscle nerve stimulation, were found between motoneurons innervating the same muscle or synergistic muscles, but never between motoneurons innervating antagonistic muscles. These potentials were finely graded in response to a series of increasing stimuli and resistant to high frequency (50Hz) stimulation. CONCLUSIONS: These results indicate that excitatory connections, with certain specificity of spatial and temporal distribution, occur in the spinal motoneurons. It is also suggested that electrical coupling should be involved in these connections and this mechanism should improve the excitability of the motoneurons in the same column. PMID- 12899398 TI - Protective effect of melatonin on neural cells against the cytotoxicity of oxyradicals. AB - OBJECTIVE: To investigate the exact mechanism of melatonin to prohibit the apoptosis of neural cells induced by various kinds of cytotoxic agents. METHODS: We used the methods of phase contrast microscopy, MTT assay and hoechst dye staining to check this mechanism in SKNSH and U251 cell lines. RESULTS: Both 2 mmol/L H2O2 and 0.5 micromol/L amyloid beta-protein (Abeta) induce these two cell lines die via apoptosis. Either melatonin or glutathione can significantly protect both cell lines. The protective effect of 10 micromol/L melatonin is as same as that of 60 micromol/L glutathione. CONCLUSION: Melatonin can partly inhibit the cytotoxicity of H2O2 and Abeta through its role as a free radical scavenger. PMID- 12899399 TI - Chromosome studies in 51 couples with spontaneous abortions. PMID- 12899400 TI - The role of hysterectomy in the therapy of gestational trophoblastic tumor. AB - OBJECTIVE: To evaluate the role of hysterectomy for patients with gestational trophoblastic tumor. METHODS: We retrospectively analyzed 68 cases of gestational trophoblastic neoplasia treated by hysterectomy from 1985 to approximately 1997 at PUMC hospital. Thirty-eight cases were diagnosed of choriocarcinoma and 30 were invasive mole. RESULTS: Twenty-three elder patients who didn't desire to preserve fertility were selected for hysterectomy after shorter courses of chemotherapy, 22 of them had a complete remission (95.6%), the total average courses of chemotherapy was 4.2. Of twenty-seven chemorefractory cases who were suspected of a refractory isolated lesion in the uterus, delayed hysterectomy as an adjunct to chemotherapy was performed, 20 of them got a complete remission (71.1%), the total average courses of chemotherapy were 9.4. Emergency hysterectomy is indicated in 18 patients with uterine perforation or life threatening hemorrhage, 17 cases had a complete remission (94.4%), the total average courses of chemotherapy were 7.6. CONCLUSION: Although the development of effective chemotherapy has resulted in improved survival of patients with gestational trophoblastic tumor, hysterectomy remains an important adjuncts in the treatment of a selected subset of patients; in order to operate more completely and prevent recurrence, it's better to perform extended hysterectomy for the indicated patients. PMID- 12899401 TI - A retrospective study of bilateral alveolar bone grafting. AB - OBJECTIVE: To evaluate the treatment results of bilateral alveolar bone grafting (BABG) in patients with bilateral complete clefts of lip and palate. METHODS: A retrospective study was performed in 66 bilateral complete cleft lip and palate patients who received the procedure of BABG, among them 15 were primary BABG and 51 were secondary BABG. The patients were further divided into three groups according to age and eruption stage of the canine at the time of surgery. The result of BABG was evaluated on the radiographs. RESULTS: (1) The overall success rate of BABG was 75.0%, with 83.3% and 72.5% for primary and secondary BABG respectively; (2) The marginal bone level was found to be significantly higher in the youngest age group than in the other groups both for primary and secondary BABG; (3) For both primary and secondary BABG, Group C (patients' age more than 16 years) had the least optimal success rate, with 66.7% and 65.1% respectively. CONCLUSION: Simultaneous primary palate repair and BABG is safe and feasible procedure for treating unoperated bilateral complete cleft lip and cleft palate patients. For both primary and secondary BABG, significantly better results can be achieved if the operation is performed before eruption of the canine. PMID- 12899402 TI - Congenital choledochal cyst--report of 56 cases. AB - OBJECTIVE: The aim of this paper is to describe the mechanism, clinical manifestation, diagnosis and surgical management of congenital choledochal cyst. METHODS: From 1984 to 1997, 56 cases of congenital choledochal cyst were reviewed. Among them, 8 patients were male, 48 patients were female, the age ranges from 12 to 50 years old with an average of 26.3. RESULTS: The main clinical manifestation includes jaundice, abdominal pain and abdominal mass. All of them were confirmed by ultrasonography, while 39 cases were performed ERCP and PTC with the same diagnosis. Fifty-one patients were performed cystectomy and hepatojejunostomy, two cases were performed cystjejunostomy because of diffused angioma and severe hemorrhage respectively, external drainage was performed in one case with emergent cholangitis, the other two cases were reported malignancy through biopsy and operation was abandoned. CONCLUSIONS: Ultrasound diagnostics is essential to accurately diagnose the cyst, preoperative ERCP is helpful for differentiating pancreatic duct from bile duct, while MRCP is a reliable method; cystectomy and cholangiojejunostomy is recommended, laparoscopic procedure is becoming more and more accepted. PMID- 12899403 TI - Endomyocardial fibrosis in China. AB - PURPOSE: To introduce the epidemical, pathological, and clinical characteristics as well as the diagnostic and therapeutical experiences of endomyocardial fibrosis (EMF) in China. DATA SOURCES: A CMBdisc search was done of the Chinese language literature published from January 1983 through June 1997 about EMF and/or restrictive cardiomyopathy. A manual search was then done for other contributions, including abstracts, between January 1965 and June 1997. RESULTS: Eighty-seven Chinese cases of EMF were collected in this paper. There were 49 men and 38 women, with a mean age of 28 +/- 13 years (range, 8 to 68 years). The distribution of the cases is mainly in the south of China. Combined right and left ventricular disease occurs in 48 percent of cases, with pure right ventricular involvement occurring in 12 percent and pure left ventricular involvement in the remaining 10 percent of patients who are examined postmortem. The diagnosis of EMF was confirmed in 21 cases at autopsy, and in 66 cases by echocardiography, angiocardiography, and/or endomyocardial biopsy which showed the characteristic changes. Clinically, right-sided disease is the commonest variety. Endocardiectomy and tricuspid (n - 7) or mitral (n - 1) valves replacement have been performed in 8 patients. There were 2 operative deaths. Six patients had a satisfactory recovery postoperatively and living well in the follow-up duration. CONCLUSION: EMF has been diagnosed clinically and confirmed at necropsy in a number of cases in the south of China. The etiology, incidence and epidemiology are still unknown. The pathological and clinical features are similar to those in tropical areas, but right ventricular involvement is the commonest type in our country. PMID- 12899404 TI - Sacral fractures. AB - OBJECTIVE: To delineate the clinical spectrum and treatment choice of sacral fractures. METHODS: In this series, 39 sacral fractures were retrospectively reviewed and classified utilizing Denis' classification. There were 21 Zone I fractures, 6 Zone II fractures and 12 Zone III fractures. Neurological deficits were present in seven patients. Thirty seven patients were treated conservatively and two underwent surgical management. RESULTS: Thirty eight patients were followed up for three months to 19 years. Thirty three have recovered, four improved, and one remained disabled. CONCLUSION: The treatment of sacral fractures requires assessment of pelvic stability and existing nerve injury. The patients with pelvic ring instability and neurological deficits should be treated with fracture reduction and stability reconstruction. When the patients with pelvic fracture are complicated with neurological deficits, sacral fracture should be first suspected. Once the diagnosis of sacral fracture is made, fracture reduction should be indicated. Conservative treatment usually permits satisfactory results. PMID- 12899405 TI - The value of 99mTc-MIBI myocardial perfusion spect imaging in detecting coronary artery disease in patients with valvular disease before operation. AB - OBJECTIVE: The aim of this study was to detect coronary artery disease using 99mTc-MIBI myocardial perfusion imaging in patients with valvular disease. METHODS: Thirty patients with valvular disease confirmed by echocardiography underwent 99mTc-MIBI myocardial perfusion imaging using multiSPECT 1h after stress test (exercise, dipyridamole or dobutamine test) and were performed coronary angiography within 1 month before valvular operation. RESULTS: For 29 out of the 30 patients, the results of 99mTc-MIBI myocardial perfusion imaging were similar with those of coronary angiography, the concordance rate was 96.7% and the negative predictability was 100%. CONCLUSION: 99Tc-MIBI myocardial perfusion imaging is a reliable non-invasive method for detecting coronary artery disease in patients with valvular disease and so as to draw up suitable operation programs for them. PMID- 12899406 TI - Pentacapsula neurophila sp.n. (Multivalvulida) from the central nervous system of striped trumpeter, Latris lineata (Forster). AB - Striped trumpeter, Latris lineata, are being experimentally cultured by the Tasmanian Aquaculture and Fisheries Institute at Taroona, Tasmania. Fish surviving over 30 days post-hatching have frequently developed nervous aberrations associated with a severe granulomatous meningoencephalomyelitis. The myxozoan parasite Pentacapsula neurophila sp.n. was revealed as the parasite causing the disease in the striped trumpeter juveniles. Measurements made of isolated spores indicated that the organism was distinct from all previously described Pentacapsula species. This is the first report of a marine myxozoan parasite of the genus Pentacapsula in Australian waters. PMID- 12899407 TI - Non-specific immune response of turbot, Scophthalmus maximus (L.), experimentally infected with a pathogenic Vibrio pelagius. AB - The effect of a pathogenic Vibrio pelagius, isolated during a mass mortality of turbot larvae, on the non-specific immune response of turbot, Scophthalmus maximus (L.), macrophages was studied both in vitro and in vivo. The in vitro treatment of head kidney (HK) macrophages with viable V. pelagius caused a significant inhibition of the chemiluminescence (CL) response in comparison with untreated macrophages, while incubation with heat-killed bacteria did not affect this response. In vivo, the intraperitoneal injection of V. pelagius resulted in a significant inhibition of the CL response in infected fish at days 1 and 4 post infection compared with the control fish response. The HK macrophage nitric oxide (NO) production was enhanced by in vitro incubation with intermediate doses of viable V. pelagius (5 x 10(3) and 5 x 10(4) bacteria mL(-1)) and higher doses of the heat-killed bacteria (5 x 10(4)-5 x 10(6) bacteria mL(-1)). In both cases, the NO inhibitor N-omega -nitro-L-arginine was capable of down-regulating the specific NO induction caused by incubation with the bacterial treatments. In contrast, incubation with ECPs at higher doses caused a reduction in NO production. In vivo, a significant enhancement in NO production was also observed in macrophage supernatants at day 10 post-infection. Lysozyme concentration in the serum was also significantly increased in the experimentally infected fish at days 4 and 10 post-injection. In addition, viable V. pelagius and its ECPs significantly reduced HK macrophage viability in vitro, whereas no significant differences in viability were observed during the incubation with heat-killed bacteria. As NO production was enhanced in the experimentally infected fish, the inhibitory effect of the NO donor, S-nitroso-acetyl-penicillamine (SNAP), was tested in vitro in a cell-free assay. The results showed that growth of V. pelagius was significantly inhibited using SNAP at a high concentration (1 mM). PMID- 12899408 TI - Protection against atypical furunculosis in Atlantic halibut, Hippoglossus hippoglossus (L.); comparison of a commercial furunculosis vaccine and an autogenous vaccine. AB - Atlantic halibut, Hippoglossus hippoglossus (L.), was shown to be sensitive to infection by three different isolates of Aeromonas salmonicida ssp. achromogenes in pre-challenge tests using intraperitoneal (i.p.) and intramuscular (i.m.) injections as well as bath challenges. A commercial furunculosis vaccine, Alphaject 1200, and an autogenous vaccine, AAS, based on the challenge strain, induced immune protection as shown in challenge tests 8 weeks post-immunization. The survival rate of vaccinated fish after i.p. challenge was 100%, whereas mortality of control fish was 61%. Employing i.m. challenge, relative percentage survival induced by the furunculosis vaccine and the AAS vaccine was 47 and 44, respectively. Mortality of i.m. injected controls was 68%. Vaccinated fish behaved normally following vaccination but the weight gain was significantly reduced in vaccinated fish 8 weeks post-vaccination compared with control fish receiving phosphate-buffered saline. At the same time, intra-abdominal adhesions were observed in fish injected with either of the two vaccines or adjuvant alone. Antibody response against A. salmonicida ssp. achromogenes was detected in sera from fish receiving either vaccine. PMID- 12899409 TI - A single-dose pharmacokinetic study of oxolinic acid and vetoquinol, an oxolinic acid ester, in cod, Gadus morhua L., held in sea water at 8 degrees C and in vitro antibacterial activity of oxolinic acid against Vibrio anguillarum strains isolated from diseased cod. AB - The pharmacokinetic properties of the antibacterial agent oxolinic acid and vetoquinol, the carbitol ester of oxolinic acid, were studied after intravenous (i.v.) and oral (p.o.) administration to 100-150 g cod, Gadus morhua L., held in sea water at 8 degrees C. Following i.v. injection, the plasma drug concentration time profile showed two distinct phases. The distribution half-life (t1/2alpha) was estimated at 1.3 h, the elimination half-life (t1/2beta) as 84 h and the total body clearance (Cl(T)) as 0.047 L kg(-1) h(-1). The volume of distribution at steady state, Vd(ss) was calculated to be 5.5 L kg(-1), indicating good tissue penetration of oxolinic acid in cod. Following p.o. administration of oxolinic acid or vetoquinol, the peak plasma concentrations (C(max)) of oxolinic acid and the time to peak plasma concentrations (T(max) were estimated to be 1.2 and 2.5 microg mL(-1) and 24 and 12 h, respectively. The bioavailabilities of oxolinic acid following p.o. administration of oxolinic acid and vetoquinol were calculated to be 55 and 72%, respectively. The in vitro minimum inhibitory concentration (MIC) values of oxolinic acid against three strains of Vibrio anguillarum isolated from diseased cod were 0.016 microg mL(-1) (HI-610), 0.250 microg mL(-1) (HI-618) and 0.250 microg mL(-1) (HI-A21). Based on a MIC value of 0.016 microg mmL(-1) a single p.o. administration of 25 mg kg(-1) of oxolinic acid maintains plasma levels in excess of 0.064 microg mL(-1), corresponding to four times the MIC-value, for approximately 12 days. The analogous value for a single p.o. dose of 25 mg kg(-1) of oxolinic acid administered as vetoquinol was 13 days. PMID- 12899410 TI - Putative virulence factors of atypical Aeromonas salmonicida isolated from Arctic charr, Salvelinus alpinus (L.), and European grayling, Thymallus thymallus (L.). AB - Atypical Aeromonas salmonicida (AAS) causes generalized lethal infections in farmed Arctic charr, Salvelinus alpinus (L.), and European grayling, Thymallus thymallus (L.), and is thus a serious threat for culture of these fish species. Virulence factors were studied among isolates of AAS from Arctic charr (n = 20), European grayling (n = 19) and other fish species (n = 20), of which 48 were of Finnish and 11 of Swedish origin. All isolates produced an A-layer. Extracellular products (ECP) of the AAS isolates did not produce detectable gelatinase and caseinase activity in test assays. Analysis of the same ECP preparations with substrate sodium dodecyl sulphate-polyacrylamide gel electrophoresis showed weak proteolytic activity, indicating the different sensitivity of the detection methods used. The ECP from AAS isolates showed low cytotoxic activity against cultured cells. However, the ECP did not induce mortality in challenged Arctic charr. The results suggest that toxic components, like ECP, secreted by the bacterium may not be the major virulence factor in AAS-infection in Arctic charr and European grayling, and hence the pathogenesis also differs from the pathogenesis of AAS-infection in Atlantic salmon, Salmo salar L. PMID- 12899411 TI - Viral encephalopathy and retinopathy of Dicentrarchus labrax (L.) farmed in fresh water in Greece. PMID- 12899412 TI - Immunostimulatory CpG oligodeoxynucleotides increase resistance against amoebic gill disease in Atlantic salmon, Salmo salar L. PMID- 12899413 TI - Pasteurellosis in Atlantic salmon, Salmo salar L.: immunohistochemistry of the naturally-occurring disease. PMID- 12899414 TI - Age and gender differences in cardiac reactivity and subjective emotion responses to emotional autobiographical memories. AB - This research reports age and gender differences in cardiac reactivity and subjective responses to the induction of autobiographical memories related to anger, fear, sadness, and happiness. Heart rate (HR) and subjective state were assessed at baseline and after the induction of each emotion in 113 individuals (61 men, 52 women; 66% European American, 34% African American) ranging in age from 15 to 88 years (M = 50.0; SD = 20.2). Cardiac reactivity was lower in older individuals; however, for anger and fear, these age effects were significantly more pronounced for the women than the men. There were no gender differences in subjective responses, however, suggesting that the lower cardiac reactivity found among older people is dependent on gender and the specific emotion assessed. PMID- 12899415 TI - Corticolimbic mechanisms in emotional decisions. AB - Midline frontolimbic networks are engaged in monitoring simple actions. They may also provide evaluative control for more complex decisions. Subjects read a trait descriptive word and responded either "yes" or "no" within 1,500 ms whether it was self-descriptive. By 300 ms, an electrophysiological discrimination between good and bad words was seen over centromedial regions of the frontal lobe for both friend and self-decisions. By 350 ms, an interaction effect between evaluation and endorsement appeared, and by 500 ms, activity specific to self evaluation was seen in both anterior and posterior midline sites. An evaluative decision thus begins by recruiting motivational and semantic influences within limbic networks, and these influences appear to shape the development of the decision within multiple neocortical regions. PMID- 12899416 TI - What do facial expressions convey: feeling states, behavioral intentions, or action requests? AB - Emotion theorists assume certain facial displays to convey information about the expresser's emotional state. In contrast, behavioral ecologists assume them to indicate behavioral intentions or action requests. To test these contrasting positions, over 2,000 online participants were presented with facial expressions and asked what they revealed-feeling states, behavioral intentions, or action requests. The majority of the observers chose feeling states as the message of facial expressions of disgust, fear, sadness, happiness, and surprise, supporting the emotions view. Only the anger display tended to elicit more choices of behavioral intention or action request, partially supporting the behavioral ecology view. The results support the view that facial expressions communicate emotions, with emotions being multicomponential phenomena that comprise feelings, intentions, and wishes. PMID- 12899417 TI - Separating emotion and motivational direction in fear and anger: effects on frontal asymmetry. AB - State effects on frontal alpha electroencephalograph asymmetry (ASY) are thought to reflect approach and withdrawal motivational tendencies. Although this motivational direction model has inspired a large body of research, efforts to disentangle influences of emotion (EMO) and motivational direction (MOT) on ASY are rare. The authors independently manipulated EMO (fear and anger) and MOT (approach and withdrawal) in a between-subjects design. Irrespective of MOT, anger led to greater changes toward relative left frontal activation (LFA) than did fear. Conversely, higher ratings of negative valence were associated with greater changes toward LFA in withdrawal but with greater changes toward relative right frontal activation in approach. Results are discussed within a model based on behavioral inhibition system-behavioral activation system theory. PMID- 12899418 TI - It takes one to know one: interpersonal sensitivity is related to accurate assessments of others' interpersonal sensitivity. AB - Interpersonal sensitivity (emotional and social) is the ability to accurately assess others' abilities, states, and traits from nonverbal cues. The authors predicted that individuals' interpersonal sensitivity would be related to accurate judgments of friends' interpersonal sensitivity. Fifty participants were recruited, each bringing a friend to participate in performance-based, self report, and other-rating measures of emotional and social sensitivity. Interpersonal sensitivity was related to accurate judgments of others' interpersonal sensitivity (the "it-takes-one-to-know-one effect"). Neither gender nor acquaintanceship was directly related to accurate judgments of interpersonal sensitivity, nor did either variable moderate the it-takes-one-to-know-one effect. PMID- 12899419 TI - Autobiographical memory specificity and affect regulation: an experimental approach. AB - This study investigated J. M. G. Williams's (1996) affect-regulation hypothesis that level of specificity of autobiographical memory (AM) is used to minimize negative affect. It was found that a negative event leads to more reports of subjective stress in high- as compared with low-specific participants. Also, afterward, high-specific participants rated their unprompted memories for the event as more unpleasant. The results indicate that, relative to high specificity, being less specific in the retrieval of AMs is associated with less affective impact of a negative event. Results are discussed within the affect regulation model. It is suggested that future research take a more functional perspective on AM specificity. PMID- 12899420 TI - The role of forensic science in the trial of Harold Shipman. PMID- 12899421 TI - The Shipman inquiry: a progress report. PMID- 12899422 TI - Death certification and Dr. Shipman. PMID- 12899423 TI - Depression, homicide and diminished responsibility: new Scottish directions. AB - In a recent Scottish Appeal Court opinion (Kim Louise Scarsbrook or Galbraith v. Her Majesty's Advocate, 2001) it was successfully argued by the appellant that her conviction of murder was unsound inter-alia on the basis of overly restrictive pre-existing definitions of diminished responsibility in Scottish law resulting in unduly narrow directions being given by the trial judge to the jury in her case. We felt it timely to present a revised overview of the defence of diminished responsibility in Scotland and to consider the issues surrounding its applicability in cases of clinical depression. The psychiatric literature regarding depression and homicide is reviewed. PMID- 12899424 TI - Childhood homicide: a 1990-2000 retrospective study at the Institute of Legal Medicine in Lausanne, Switzerland. AB - The authors retrospectively reviewed all homicides of children under 15 years of age referred to the Institute of Legal Medicine in Lausanne from January 1990 to December 2000. We report on 41 cases during this eleven-year period. Among those, 17 newborns and 24 older children, a higher proportion of males was found. Regarding newborn homicides, when the identity of the mother was known she was always identified as the perpetrator. With non-newborn homicides, the majority of the victims were killed at home and knew the offender. Two common profiles of these homicides could be determined. In the first scenario, one of the parents shot all the children and committed suicide afterwards. The second profile is defined as 'fatal child abuse' and concerned younger victims whose cause of death was the result of a cranio-cerebral trauma from battering or shaken baby syndrome. PMID- 12899425 TI - Undetermined drowning. AB - Drowning is one of the leading causes of death when the manner of death remains undetermined. In the present study, we examined the epidemiological and medico legal profile of 276 undetermined deaths (M:F=3.4:1; mean age 41.9+/-16.0 SD) among 1,707 consecutive bodies found in water and autopsied at the Department of Forensic Medicine, University of Helsinki, from 1976 to 2000. We also describe the differences between the police investigator's initial opinion and the forensic pathologist's death certification, and the different approaches among forensic pathologists when determining the cause of death. There was considerable variation among individual pathologists in the percentage of deaths considered undetermined but these differences were not significantly related to their level of training. Medico-legal training should focus on a standardised diagnostic approach to borderline cases, in which essential factors in determining the manner of death are often ambiguous. PMID- 12899426 TI - Gender-related traumatic deaths in Transkei: incidence and causes. AB - This study is unique in that it strives to unfold, perhaps for the first time, the problem of female mortality due to trauma in the Transkei region of the Eastern Cape Province of South Africa. This study was carried out in the Umtata and Ngqeleni magisterial districts which have a combined population of about 400,000. Most of the people have very few resources and have historically relied on money repatriated by migrant workers. The objective was to establish the incidence and the causes of deaths due to gender-related trauma and to formulate recommendations which could probably help prevent or reduce these deaths. The study reviewed cases of female traumatic death during the period January 1993 to December 1999 that were brought to the mortuary in Umtata General Hospital (UGH). There were 1,054 (23%) traumatic deaths recorded in females between 1993 and 1999. Of these 486 (28%) were related to motor vehicle collisions, 219 (18%) due to gunshot injuries, 152 (19%) due to stab wounds and 139 (21%) as a result of blunt trauma. The male to female ratio was 3.3:1 in traumatic deaths. In homicides the male:female ratio was 4.4:1, gunshot 4.5:1, stab 4.2:1 and blunt injuries 3.7:1. In motor vehicle collisions (MVC's) the ratio was 2.5:1. There is an increasing incidence of traumatic deaths in women. Gun shot injuries are the commonest among traumatic deaths in females. This article recommends stricter measures to protect women in the form of legislation. Social uplifting and economic support should be carried out as part of the process of social change. In this case educating the entire community is necessary to safeguard women and their future survival. PMID- 12899427 TI - Suicides in northern India: causes, methods used and prevention. AB - Suicide patterns and rates differ in various populations and cultures. It has been reported that developing countries have lower suicide rates, possibly because of a lower level of environmental stress but the Indians have a very high suicide rate (Elfawal, 1999). The choice of method used to commit suicide depends on availability of means, knowledge about lethal effectiveness, and the victim's motivation and intent. The present work was designed to investigate the different methods of self-destruction, the age and gender susceptibility to suicide, the groups particularly affected and the underlying motivating factor for such an extreme step among the North Indians. Various suggestions relating to decreasing the tensions of the modern mechanical life-style, educating the public in general and the availability, use and storage of agrochemicals have been put forward. Marriage counselling and the concept of family planning are also advocated. PMID- 12899428 TI - Ascertaining capacity to consent: a survey of approaches used by psychiatrists. AB - Doctors are increasingly criticised for imposing treatment without adequate consent. Approaches used by psychiatrists (of all grades and in all sub specialities) to assess capacity to consent in a large west London psychiatric trust were surveyed with a questionnaire. The questionnaire consisted of two case vignettes and a series of seven questions (six with forced choice yes/no answers and one with an open-ended answer). The responding psychiatrists' assessment of capacity and that judged by the research psychiatrist were compared. There was no relationship between age, sex, duration of experience in psychiatry, grade and speciality, and (i) the respondents' assessment of capacity and (ii) the research psychiatrist's assessment of capacity. The kappa concordance between the responding psychiatrists' assessment of capacity and that judged by the research psychiatrist was 0.17 (P=0.067) and 0.12 (P>0.05) for vignettes 1 and 2 respectively. Individual psychiatrists appear to variably interpret and apply the criteria for assessment of capacity to consent. PMID- 12899429 TI - A study of the investigation of death (coroner system) in Sri Lanka. AB - Sri Lanka (Ceylon) inherited the 'coroner system' of investigating death in the early nineteenth century. Unlike in England and Wales, the coroner system in Sri Lanka did not change much in the last century. This study, the first of its kind, was performed for a period of three months in 1995 to analyse the causes and circumstances of deaths reported for inquest, and the number of autopsies performed. Of the 868 cases reported to the Inquirer, the circumstances were not determined in 94 cases at the initial inquest proceedings. Of the other 774 cases, 454 (58.7%) were natural deaths. Autopsies were performed on 44.5% of natural deaths, 58.2% of accidental deaths, 96% of road traffic accidents, 44.9% of suicides and 81.6% of homicides. Coronary heart disease was the leading cause of natural death (33.9%). Head injuries were responsible for 31.8% of accidental deaths while burns accounted for 24.5% of accidental deaths and 46.3% of suicides. There were 38 cases (4.9%) of homicide of which autopsies were performed in 31. There is an urgent need to reform the century-old laws relating to inquest procedures in the country. PMID- 12899430 TI - Blood analysis by headspace gas chromatography: does a deficient sample volume distort ethanol concentration? AB - This study was prompted by a recent judgment in the Royal Courts of Justice (Gregory v. Director of Public Prosecutions, 2002) in a case of driving a motor vehicle after consuming too much alcohol (Road Traffic Act 1988). An expert witness for the defence alleged that a deficient volume of blood in the tube sent for analysis meant an excess amount of sodium fluoride (NaF) preservative, which would increase the concentration of ethanol, determined by headspace gas chromatography (HS-GC), owing to a salting-out effect. The prosecution did not produce expert evidence to rebut this argument and the drunk driving suspect was acquitted. A small volume of blood and excess sodium fluoride might have increased the concentration of ethanol in the air-space in the tube sent for analysis but this does not mean that the result of the HS-GC analysis would be higher. This follows because prior to analysis an aliquot of blood is removed and diluted (approximately 10 times) with n-propanol as the internal standard. The dilution lowers the concentration of NaF in the blood and for quantitative analysis the ratio of the ethanol to n-propanol response is measured. The use of a ratio also helps to compensate for any salting-out effect of ethanol. Our experiments showed that a deficient volume of blood and excess NaF actually lowered the concentration of ethanol by 2-3% compared with heparinised blood. Seemingly, n-propanol (n-PrOH) a 3-carbon straight chain alcohol is salted out slightly more effectively than the 2-carbon ethanol (EtOH) causing a lower peak area ratio (EtOH/n-PrOH) and a lower apparent concentration of ethanol. In a separate study, we showed that the concentration of ethanol was lowered even more when a 4-carbon alcohol (t-butanol) was used as the internal standard. PMID- 12899431 TI - First compulsory admissions under part II of the Mental Health Act 1983: a retrospective five-year study. AB - The aim was to explore what happened during a five-year period to a group of patients with no previous psychiatric admissions, who were detained for the first time in a psychiatric hospital under part II of the Mental Health Act 1983. Subjects (n=189) who were detained under Section 2 or 3 in 1996 and 1997 in Queen Elizabeth Psychiatric Hospital, Birmingham were identified from computerised inpatient admission records. The mean age was 40.0 years (range 21-65). Nearly half of the cohort had a diagnosis of psychoses. Asians had the highest proportion of psychoses (64%) compared to African-Caribbean patients (48%) and White patients (41%). Almost half of the sample had a further admission, with Asian and African-Caribbean patients more likely to be readmitted than their white counterparts. Nearly a third of the cohort had a further compulsory detention. On re-admission, African-Caribbean subjects were more likely to be detained compared to Asian and White subjects. Fifty per cent had a further compulsory admission within a year of the index admission. The high number of psychoses in the Asian group may be due to their psychosis being more serious at presentation as a result of denial of illness and an increased co-morbidity with drug use in Asian men. PMID- 12899432 TI - Psychiatric support to magistrates' courts. AB - There are many problems associated with magistrates' courts obtaining psychiatric advice when faced with defendants with mental health problems. Often the disposal of such individuals is less than ideal because of the unavailability of expert advice. In order to overcome these difficulties in Hampshire, a protocol has been developed to guide the court towards other sources of expert psychiatric advice as an alternative to the traditional route of using a consultant psychiatrist. By using existing court diversion schemes, prison mental health teams and care co ordinators in community mental health teams, it is hoped that this will result in a more timely and reliable flow of psychiatric information. Not only will this provide an improved service to the courts but the mentally disordered defendant should benefit from a better informed bench when they are considering his/her disposal. PMID- 12899433 TI - Child sexual abuse in Singapore with special reference to medico-legal implications: a review of 38 cases. AB - There is a paucity of studies on child sexual abuse in Asia. This study reviews the trends of child sexual abuse in Singapore and discusses the medico-legal considerations in the identification, assessment and management (including forensic implications) of such cases. It is a retrospective case review of 38 consecutive cases of child sexual abuse seen at a child guidance clinic in Singapore. Our study showed that most of the children were young (74% below age 9) and female (78.9%) with perpetrators who are males and usually known to the victims. There is a need to have integrated protocols to streamline assessment and reduce the need for repeated questioning. Adequate support of the victims in the form of specialised handling during the Court proceedings and in the use of new technology (such as video conferencing) for obtaining the child's testimony may be necessary. A child's fitness to testify is determined by the credibility and competence of the child. Repressed memories are of questionable validity. PMID- 12899434 TI - Sudden unexpected death of a methamphetamine abuser with cardiopulmonary abnormalities: a case report. AB - A 51-year-old man suddenly collapsed while being arrested. He was an abuser of methamphetamine (MP) with a history of previous convictions. The blood MP concentration at autopsy was below the lethal level. The heart revealed hypertrophy and endocardial thickening of the bilateral ventricles. On microscopic examination, the typical findings of hypertrophic cardiomyopathy were not seen, however endocardial thickening with increasing elastic fibres was seen in all the ventricles and in the atria. In the conduction system, the AV node artery with luminal narrowing was longitudinally compressed by circumferential tissue. Severe interstitial fibrosis and small scar formation of the superiorventricular septum were also seen. In addition, diffuse hypertensive pulmonary arteriopathy was seen in the lung parenchyma. We surmise that the AV node artery had been chronically compressed by the circumferential architecture, due to a chronic increase in intracardiac pressure. We believe that the pathological lesions noted in both the cardiac conduction system and the lung in our case may be a complication of MP abuse, and that an MP abuser with such lesions could easily die suddenly upon experiencing emotional stress. PMID- 12899435 TI - Law and science. Civil law. Medical negligence. PMID- 12899436 TI - The man who teached sciences by travelling around the world. PMID- 12899437 TI - Immunolocalization of S100A2 calcium-binding protein in cartilage and bone cells. AB - S100A2 protein, a Ca2+ binding protein, was investigated by immunocytochemistry in the epiphyseal cartilage and bone cells of growing rats, and in primary cultures of osteoblasts. S100A2 was detected in the chondrocytes and in the extracellular cartilage matrix. In the later however, its presence only in the calcifying areas of the epiphyseal cartilage suggests that it could be involved in the process of calcification of cartilage. PMID- 12899438 TI - Elemental maps in human allantochorial placental vessels cells. 3. 5 hydroxytryptamine effects. AB - The membrane potential, a regulator of vascular tone, is a function of the physiological activities of ionic channels (particularly, K+ and Ca2+ channels in these cells). These channels regulate the ionic distribution into these cells. Micro-particule induced X-ray emission (PIXE) analysis was applied to determine the ionic composition of vascular smooth muscle cells (VSMCs) and of vascular endothelial cells (VECs) in the placental human allantochorial vessels in a physiological medium (Hanks'solution) modified by the addition of a chemical stimulus: 5-hydroxytryptamine (5-HT), an activator of the voltage-sensitive Ca2+ channels. In VSMCs (media layer), the addition of 5-HT induced no modification of the Na, K, Cl, P, S and Ca concentrations but increased Mg concentration. In endothelium (VECs) 5-HT addition implicated an increase of the K, S, Ca concentrations, the concentration of the other ions remained constant. In VECs, Ca and K increase is due to open of L-type voltage-dependent Ca2+ channels and of K(Ca) channels. 5-HT induces also a secretion of endothelium hyperpolarizing factors which implicate decrease of [Ca2+]i in VSMCs opposite to a direct increase by 5-HT. Increase in [Mg2+]i may be due to activation of the Ca/Mg exchanger. PMID- 12899439 TI - Acute porphyrias in the Argentinean population: a review. AB - The porphyrias are a group of inherited metabolic disorders of heme biosynthesis which result from a partial deficiency in one of its seven specific enzymes, after its first and rate limiting enzyme, delta-aminolevulinic acid synthetase. They can be classified on the basis of their clinical manifestations into cutaneous, acute and mixed disorders. Acute intermittent porphyria (AIP) is the most common type of hepatic acute porphyrias, inherited as an autosomal dominant trait, caused by a defect in the gene which codifies for the heme enzyme porphobilinogen deaminase. Its prevalence in the Argentinean population is about 1:125,000. A partial deficiency in another enzyme, protoporphyrinogen oxidase, produces variegate porphyria (VP), the second acute porphyria most frequent in the Argentinean population (1:600,000). Here, we review all the mutations we have found in 46 AIP and 9 VP unrelated Argentinean patients. To screen for mutations in symptomatic patients, we have proposed a geneticresearch strategy. PMID- 12899440 TI - Assessment of the effect of Punica granatum (pomegranata) on the bioavailability of the radiopharmaceutical sodium pertechnetate (99mTc) in Wistar rats. AB - The many desirable characteristics of technetium-99m (99mTc) have stimulated the development of labeling techniques for different molecular and cellular structures. It is generally accepted that a variety of factors other than disease can alter the bioavailability of radiopharmaceuticals and one such factor is the drug therapy. The use of medicinal plants has increased in the last decades all over the world. Punica granatum (pomegranata) is used as food or as medication in folk medicine for antiviral, anthelmintic, antifungal, antibacterial and antimicrobial activity. We have studied in rats, the effect of the medicinal plant Punica granatum on the bioavailability of the radiopharmaceutical 99mTc sodium pertechnetate (Na(99m)TcO4). The infusion of pomegranata was administered by intragastric via into Wistar rats during seven days. After that, the animals received by ocular plexus via, 0.1 ml of the Na(99m)TcO4 (3.7MBq) and the animals were rapidly sacrificed after 5, 20 and 40 min. The organs were isolated (brain, heart, thyroid, liver, lungs, kidneys, stomach, testis, intestines, pancreas, spleen, bladder, muscle and bone), the radioactivity determined in a well counter, the percentages of radioactivity (%ATI) in the organs were calculated and statistical analyses were performed by Wilcoxon test (p < 0.05). The results have shown a significant (p < 0.05) increase of the activity of the Na(99m)TcO4 in spleen, heart, stomach, liver, stout bowel, pancreas, lungs and testis at 5 min. Twenty minutes after the administration of the radiopharmaceutical, the analysis of the results reveals a significant (p < 0.05) increase of the %ATI in heart, stomach, femur, pancreas, lungs and kidneys. Forty minutes after the administration of the Na(99m)TcO4, the results show a significant (p < 0.05) increase in spleen, brain, heart, stomach, liver, stout bowel, muscle, femur, lungs, pancreas, kidneys and testis. These results can be justified by therapeutic effect of this extract and/or by generation of active metabolites capable to interfere with the biodistribution of the studied radiopharmaceutical. PMID- 12899441 TI - Evaluation of glycogen loss in human liver transplants. Histochemical zonation of glycogen loss in cold ischemia and reperfusion. AB - To find a prognosis model of human liver transplant, we evaluate 62 surgical biopsies for the loss of glycogen and its variations in relation to cold ischemia, reperfusion, lobular zonation and donor's ages. We applied univariate, multivariate and discriminant analysis and logistic regression. There was a clear lobular zonation of glycogen during cold ischemia and at reperfusion. During cold ischemia, the mean loss was 48% in periportal zones and 74% in pericentrilobular zones. At reperfusion, it was in the range of 60% in periportal zones and 95% in pericentrilobular zones. It was observed in 64% of the grafts for an ischemia time less than 10 hr and in 82% of the grafts for an ischemia time of 10 hr or more. It was increased by 90% at reperfusion with pericentral predominance. Donors' age was an aggravating factor of glycogen loss beyond 28 years of age. In conclusion, in periportal zones, mean global glycogen depletion was about 54% during cold ischemia and reperfusion. It decreased by 90% at reperfusion with pericentral predominance. Logistic regression has allowed modelization of cold ischemia and reperfusion. PMID- 12899442 TI - Possible functional implications of aquaporin water channels in reproductive physiology and medically assisted procreation. AB - Spermatogenesis, the maturation of spermatozoa and their concentration and storage in the seminiferous vessels are associated with considerable fluid secretion or absorption in the male reproductive tract. These fluid movements are in total agreement with the presence of multiple aquaporin (AQP) water channel proteins in germ cells and other tissues within the male reproductive tract. A series of functions of prime importance have already been hypothesized for aquaporins in the physiology of male reproduction. Aquaporins could be involved in the early stages of spermatogenesis, in the secretion of tubular liquid and in the concentration and storage of spermatozoa in the epididymis. In the male reproductive tract, alterations in the expression and functionality and/or regulation of aquaporins have already been demonstrated to be at the basis of forms of male infertility. Indeed, rats with reduced reabsorption of seminiferous fluid in the efferent ducts have been shown to be sub-fertile or infertile. Functions have also been suggested in the fertilization process, where aquaporins may play a role in maintaining osmotic homeostasis in gametes during fertilization. Aquaporins have also been suggested to mediate water movement into antral follicles and to be the pathway for transtrophectodermal water movement during cavitation. Aquaporins are the subject of considerable technological interest for cryopreservation used in medically assisted procreation, as they could be the molecular pathway by which water and/or solutes move across the plasma membrane during the process of freezing/thawing gametes and embryos. Indeed, artificial expression ofAQP3 has been showed to improve the survival of mouse oocytes after cryopreservation. PMID- 12899443 TI - Induction of melatonin synthesis in Tetrahymena pyriformis by hormonal imprinting -a unicellular "factory" of the indoleamine. AB - Melatonin is present in Tetrahymena and its synthesis can be enhanced by pretreatment (imprinting) with melatonin. Two days after imprinting melatonin level is elevated in the cells and more elevated in the supematant. Such a minute quantity, as 10(-12) M melatonin for 1 hour is able to provoke imprinting, however the effect is more expressed using 10(-6) M. Maintenance in light conditions further elevated the amount of melatonin in the cells and supematant alike, related to the melatonin content of cells kept in darkness. The experiments call attention to the light-sensitivity of imprinting-provoked melatonin production in Tetrahymena and to the possibility of using this property for important physiological functions in higher grades of phylogeny. PMID- 12899444 TI - Evaluation of serum levels of cytokines and intercellular adhesion molecule-1 (ICAM-1) in astrocytic tumours. AB - Soluble form of intercellular adhesion molecules (sICAM) are increased in serum of many inflammatory diseases and tumours: the expression of such molecules is regulated by cytokines. In the present paper serum levels of interleukin-2 (IL 2), soluble interleukin-2 receptor (sIL-2R) and sICAM-1 were evaluated in patients with glioma compared with different tumours (lung and kidney carcinoma) in order to investigate the compromise of the immune system in these malignancies and to understand the host defence mechanisms. 14 cases of astrocytomas (WHO grade II, III), 20 cases of glioblastomas (GBL, WHO grade IV), 5 cases of lung carcinoma and 6 cases of kidney carcinoma were studied; the results were compared with 15 healthy controls. IL-2, sIL-2R, sICAM-1 concentrations were assessed by an enzyme-linked immunosorbent assay (ELISA) technique. The results were analyzed by Student's t test. Our findings showed that serum levels of IL-2 and sIL-2R were increased in all cancer patients; on the contrary, sICAM-1 serum levels were not significantly increased in GBL and astrocytoma patients. The increased values of IL-2 and sIL-2R are in agreement with a depression of the immune reactivity in patients with glioblastoma and astrocytoma, as reported in literature. On the contrary the levels of sICAM-1 are unchanged in astrocytic tumours while patients with kidney carcinoma presented the higher levels and an unfavourable prognosis. PMID- 12899445 TI - Aflatoxin related occupational exposure to maize processing workers. AB - A study was undertaken on environmental mycoflora of a maize processing industry in Ahmedabad. The airborne fungal communities were isolated and identified both qualitatively by Petri-plate exposure method and quantitatively by using Andersen 6-stage viable sampler, Midget impinger and high volume samples (cone and Hexhlet for total and respirable dusts, respectively). Of all the isolates genus Aspergillus was the dominant environmental mycoflora and among all the species of Aspergillus A. flavus was the common isolates irrespective of the method applied for sample collection. Maximum number of isolates were recovered from Elevator department. From total and respirable dusts, about 56.6% and 44.4% of recovery accounted for genus Aspergillus alone. Total percentages of aflatoxin positive strains of A. flavus were 5.65% and 9.73% from total and respirable dusts, respectively. These toxigenic strains were identified on various media like CZ with 0.05% anisaldehyde, APA and CAM. Surface morphology of toxigenic strains and dust samples were carried out using SEM. PMID- 12899446 TI - Non-toxic type 2 ribosome-inactivating proteins (RIPs) from Sambucus: occurrence, cellular and molecular activities and potential uses. AB - Ribosome-inactivating proteins (RIPs) are a family of enzymes that trigger the catalytic inactivation of ribosomes. The most known member of the family is the highly poisonous two-chain ricin isolated from Ricinus communis L. Sambucus species contain a number of two-chain RIPs structurally and enzymatically related to ricin which have the noteworthy feature that, having an enzymatic activity on ribosomes, leading to the inhibition of protein synthesis, higher than ricin, they are lacking of the tremendous unspecific toxicity of ricin. Therefore, they have been called non-toxic type 2 RIPs. The most representative and studied members are nigrin b present in the bark of the common (black) elder Sambucus nigra L. and ebulin 1 present in the leaves of the dwarf elder Sambucus ebulus L. The molecular basis for the low unspecific activities of nigrin b and ebulin 1 as compared with ricin seems to be related with single changes of amino acids in the high affinity sugar binding sites of the B chains. These changes determine the intracellular traffic of these proteins and thus the cellular toxicity. Conjugation ofnigrin b or ebulin 1 to either transferrin or monoclonal antibodies provided highly active conjugates targeting cancer. Thus these non-toxic type 2 RIPs are promising tools for cancer therapy. PMID- 12899447 TI - Recent advances in catalytic peroxidase histochemistry. AB - Immunoassays have developed to become an important analytical tool in life sciences for detection of endogenous and exogenous targets. Among the most important enzyme labels horseradish peroxidase (HRP), alkaline phosphatase (AP), and beta-D-galactosidase (GAL) is HRP the smallest enzyme and plays nowadays an outstanding role. The oldest substrates are chromogens widely applied for localization of sites of peroxidase (PO) activity in histochemistry as well as for colorimetric applications. They are represented by a diversity of aromatic amines and phenols. Encouraged by development of light excitation and measuring techniques and the commercial availability of highly sensitive equipment, luminescent labels represent the most sensitive and worthwhile detection tools to date. In contrast to chromogens fluorescent labels for detection PO activity are confined only to a few substrates developed more recently. These substrates are mostly applied in histochemistry at a short time scale due to their frequently high solubility. At the long time scale sole exception is so far the tyramine based fluorochome deposition technique (more general: catalytic reporter deposition, CARD). Despite quite different staining behavior both fluorometric and product deposition related principles are based on 4-hydroxy phenylalkyl substrates. The following article reviews basic principles of peroxidatic substrate degradation processes including chromogenic and fluorescent approaches with emphasis on recent advances in development of chromogens and fluorogens for application in histology. As a result of systematic efforts towards the design of substrates, the range of classical precipitating chromogens as well as fluorescent techniques could be complimented by novel highly sensitive substrates with superior staining capabilities: a) Metal chelating 2-hydroxy benzylamines are derived from classical aniline substrates (two steps) and utilize metal catalytic effects in an efficient intramolecular way. The enzymatically yielded dark colored polycondensation products are applicable in histochemistry, in colorimetry and especially as precipitating electron opaque labels with enhanced osmiophilic properties for light and electron microscopy. b) Fluorescent 4 hydroxy-styryl derivatives are capable of oxidative selfanchoring reactions at the cellular level close to sites of PO activity. In contrast to deposition of tyramine conjugated fluorochromes an altered fluorochrome with improved fluorescence properties is furnished during oxidative crosslinking of the substrate. This results in a highly specific and photostable fluorescence response and an outstanding low background staining. Histochemical and immunohistochemical applications are presented. PMID- 12899448 TI - What should the standard of proof be in scientific misconduct proceedings relating to public health service-funded research? AB - This article discusses the nature and purposes behind the three standards of proof commonly used in the United States. It summarizes the analytical constructs or standards of review courts commonly use to determine the constitutional validity of standards of proof (as well as other procedural protections) in physician disciplinary proceedings. It applies these constructs to the context of scientific misconduct and an illustrative case, and shows that sound policy and morals as well as procedural due process and equal protection provisions of the United States and some state constitutions require the use of the clear and convincing evidence standard of proof in scientific misconduct proceedings. That standard is necessary to protect scientists from misuse of scientific misconduct charges and proceedings, entailing, as they do, vast discretion in bureaucratic officials as well as staggering costs. The imminent rule making proceedings at the federal level will provide a special opportunity to right a wrong that long has been long visited upon academic scientists throughout the United States. PMID- 12899449 TI - Experimental tendon repair: glycosaminoglycan arrangement in newly synthesized collagen fibers. AB - Changes in the macromolecular orientation and metachromasy of glycosaminoglycans (GAG) in newly synthesized and assembled collagen fibers in rat Achilles tendon after tendon excision were investigated in toluidine blue (TB)-stained preparations, based in the selective absorption of polarized light (= linear dichroism, LD) and of absorption of unpolarized light in situ. Extrinsic LD was observed microspectrophotometrically from the early phases of tendon repair onwards, although the absorption peaks in both parallel and perpendicular directions with respect to the plane of polarized light and the long axis of the collagen fibers occurred at the same wavelength, and thus differed from the pattern situation in normal adult controls. Compared to normal adult tendons, the pattern of LD in newly synthesized and assembled fibers was still not fully attained 110 days after surgical tendon removal. This incomplete recovery possibly reflected the influence of aging during the repair process. There was no correlation between LD and metachromasy. The highest absorption values for metachromatic staining occurred on the 7th day after tendon removal, at a time when LD was not intense. Treatment with hyaluronidase showed that the LD in the early stages of tendon repair was mostly due to hyaluronate whereas the LD in the later stages was due to chondroitin sulfates. The changes in LD during Achilles tendon repair were attributed to gradual modifications in the composition and macromolecular orientation of GAGs relative to the long axis of the collagen fibers. PMID- 12899450 TI - New appreciation of nitrogen balance obtained from Japanese navy data. AB - In 1943, one of the authors (Miura) worked on the nutritional study of Japanese navy sailors. Fortunately the raw data of this study were conserved in secret in the private file of Miura. Recently, another author (Hashimoto) was seeking the data concerning nitrogen balance of Japanese people. Miura, recalling the remaining of the old data; he picked up the records of nitrogen metabolism of three sailors (A, B and C). Subject A, 34 years old, showed a net nitrogen balance of 77 g of protein per day; whereas subject B, 20 years old and subject C, 15 years old showed positive nitrogen balance. The authors remarked the relationship between unabsorbed nitrogen and nitrogen balance: The unabsorbed ratio of food nitrogen was the greatest in A (29%), but in B (22%) and C (19%) showed rather low ratio. Recent study using tracers, revealed that some amino acids, especially unabsorbed leucine in the intestine, might act as regulators of nitrogen balance in adults. PMID- 12899451 TI - Light and electron microscopic radioautographic studies on macromolecular synthesis in amitotic hepatocytes of aging mice. AB - The problem on cell divisions whether cells proliferate by mitosis or amitosis has been the heated controversy among many biologists since the late 19th century. We confirmed by extensive experiments since the mid 20th century that all the cells proliferated by mitosis not by amitosis but that amitosis actually existed in some glandular cells such as hepatocytes or pancreatic acinar cells which showed only amitotic nuclear divisions without cytoplasmic division producing binucleate cells in several kinds of experimental animals. We further verified that such amitotic cells did not synthesize macromolecular compounds incorporating macromolecular precursors such as 3H-thymidine for DNA, 3H-uridine for RNA or 3H-leucine for proteins. Recent experiments at the end of 20th century using many groups of aging mice, from fetal day 19 to postnatal month 24, injected with such precursors, amitotic cells and resulting binucleate cells in the hepatocytes were detected by electron microscopic radioautography and compared to mononucleate cells. The results demonstrated that only a few hepatocytes showing amitotic nuclear division were found labelled with the 3 precursors demonstrating DNA, RNA and protein synthesis. However, the numbers of silver grains showing incorporations of labelled precursors in respective amitotic cells were very few. It was clarified that the amitotic cells did not synthesize such macromolecules as mononucleate hepatocytes did. On the other hand, more binucleate cells were found than the amitotic cells. DNA synthesis of mononucleate and binucleate hepatocyte nuclei was observed at perinatal stage and disappeared at adult stage. The labeling index of mononucleate hepatocytes was greater than that of binucleate hepatocytes. RNA and protein syntheses in karyoplasm and cytoplasm in both mononucleate and binucleate cells increased from perinatal stage, reaching the maxima at adult stage then decreased to senescent stage. Grain counts revealed that synthesized RNA and proteins were more in binucleate cells than mononucleate cells at respective aging stages. PMID- 12899452 TI - A proposal on the hard tissue remineralization in osteoarthritis of the knee joint investigated by FT-IR spectrometry. AB - An investigation was performed on hard tissues of the affected area in the cases of osteoarthritis of the knee joint by using Fourier transform infrared spectroscopy (FT-IR) to obtain the information on components and characteristics of the compounds in the hard tissues at the molecular level. Biological specimens used were: normal cortical bone, normal cancellous bone, sclerotic area of the affected site (subchondral bone), osteophyte and cartilage of the affected joint. FT-IR spectra of these specimens were measured and spectroscopic characteristics of various types of hard tissues were quantified. As the result of the analysis, signal ratio of PO4(3-) to amide I (PO4(3-)/amide I) was 0 in cartilage, and the ratio was at the highest in normal cortical bone of the ulna, being as high as 1.82. The signal ratio of the sclerotic area of the affected site was 1.69, and this was higher than the values in osteophyte (1.44) or cancellous bone of the tibia (1.61). It is inferred from these data that the degree of remineralization was considerably higher in osteoarthritis of the knee joint. On the other hand, in the osteophyte, which is frequently seen in the radiography of osteoarthritis, it was 1.44 for the attached osteophyte and 1.37 for the detached osteophyte, and these values were lower compared with the values in the sclerotic area of the affected site (1.69). From these results, it appears that remineralization of mineral components is very likely to occur under the acidic conditions in the articular region in this disease. We propose that the mechanism of hard tissue formation in the knee joint of osteoarthritis is a form of remineralization. PMID- 12899453 TI - Component analysis and growth process of nasopharyngeal calculus as revealed by Fourier transform infrared (FT-IR) spectroscopy. AB - A quite rare case of nasopharyngeal calculus in a woman in her twenties associated with the nasal discharge of pseudomonas infection was reported. As the substance was irregularly large in size, we extracted it partially by piecemeal resection using forceps and also by cracking technique using the holmium yttrium aluminum-garnet (YAG) laser, under saline irrigation and stereotactic microscopic navigator (SMN) system under endoscopic observation. The substance was firmly fixed to the pharyngeal tonsil bed. The final extract was a small piece of singly folded bandage, which is probably the focal background for calculus formation. In a cross section of calculus specimen removed during surgery, Fourier transform infrared (FT-IR) analysis revealed that a) signal ratio of methylene group (organic substance) to amide I (protein) was 21.6% at the nasal cavity side, gradually decreased toward nasal mucous membrane showing approximate 50%, b) signal ratio of amide I to P04(3-) (inorganic substance) ranged between 17.7% and 26.7% at the different sites and inside the calculus, the protein content was approximate 1/5 of the inorganic substance, and c) signal ratio of the methylene group to amide I at the nasal cavity site showed that their contents were almost equal. The quantity of the organic substance was estimated at approximate 1/2 quantity of the protein at both the central part and the part contacted with the mucous membrane. From these results, it seems that throughout the course of calculus growth, both inorganic substance and protein remain almost constant inside the calculus, while organic substance is released from the internal part of the calculus being probably formed at an early stage. PMID- 12899454 TI - Effect of tamoxifen treatment on motility related proteins in rat spermatozoa. AB - The study was undertaken to identify the effect of tamoxifen on the expression and phosphorylation of motility related proteins in the adult male rats. For this purpose, tamoxifen, at a dose of 0.4 mg/kg/day, was administered per os to the male rats for a period of 60 days. Cauda sperms, epididymal fluid and tissue proteins were extracted and analyzed by electrophoresis. Testicular tissues fixed in paraffin wax were analyzed for changes in the immunoexpression of interstitial tissue estrogen receptor alpha. Phosphorylation pattern of sperm proteins was studied in vitro after incubating with 32P-ATP. The expression of dynein and tubulin in sperms, and estrogen receptors in epididymis were analyzed by immunoblotting. Tamoxifen treatment did not alter the protein profile in the cauda sperms, epididymal fluid and tissues. Endogenous phosphorylation pattern of sperm proteins in vitro was also not affected, though it is possible that 32P incorporation observed in the 66 kDa protein could be estrogen receptor. Expression of sperm dynein, tubulin and epididymal estrogen receptors was unchanged as was the expression of testicular estrogen receptors. It was concluded that tamoxifen administration alters forward motility pattern characteristic of cauda sperm without any demonstrable change in the expression or activation of motility related proteins and the phosphorylation of the sperm estrogen receptors may be involved in the regulation of sperm motility. PMID- 12899455 TI - Approaches to the treatement of diabetes mellitus: an overview. AB - Diabetes related complications, if not treated, can be lethal. The basis of diabetes treatment is management of these complications by different approaches with the aim of providing a healthy life to diabetics. This article gives an overview of the various approaches currently in use to control hyperglycemia like pharmacological compounds and natural products. Many natural products have been used in traditional medicine, but only a few of them are discussed here. A combination therapy appears more useful for the treatment of diabetes rather than the use of a single compound. PMID- 12899456 TI - Allatostatin-like immunoreactivity in the optic lobe of the fly Sarcophaga bullata. AB - An antiserum against Diploptera allastostain 1 (Dip-AST1) was used to map the distribution of allatostain containing neurons in the optic lobes of the fly Saccrophaga bullata. Strongly immunoreacting neurons were found in two areas of the optic ganglia, namely, the medulla and the area between medulla and lobula. These cells were generally interneurons arborizing the base of the medulla. The positive reaction of specific populations of the optic lobe neurons against allatostain antiserum suggests some role for this neuropeptide in the visual physiology of the fly. PMID- 12899457 TI - So-called "overlap syndromes" in hepatology: combinations or associations of concurrent autoimmune liver and other diseases. AB - Based on present knowledge, the so-called "overlap syndromes" consisting of autoimmune liver diseases do not constitute entities in their own right. Likewise, no generally accepted marker such as an autoantibody is so far known. It would, therefore, be more correct to speak of combinations or associations of two or more concurrent diseases. Sixteen such combinations are considered here. PMID- 12899458 TI - Enhanced potential for metabolic studies using narrow-band sensitized FT-IR microspectroscopy. AB - The ability to detect and measure the relative concentration of deuterated substance found in tissue specimens is enhanced by adding a custom made-to-order 50 microm narrow MCT band that has an optimized response profile for the CD, ND and OH absorption bands. In the short range of interest it has a 2:1 signal advantage over the stock MCT detector and the steep cut off before the finger print region significantly diminishing the noise. The result is that levels of specific deuterated compounds experimentally administered to biological systems and dispersed by the host organism can be detected and measured The custom small target size narrow-band detector with increased signal and reduced noise enhances the potential for the use of FT-IR microspectoscopy as a novel way to study metabolism without the use of radioactive materials. PMID- 12899459 TI - Developmental biology meets with reproductive engineering; interdisciplinary science area as a breakthrough. AB - It has been postulated many times that different scientific topics and strategies often encounter each other, which create cutting edge research field resulting in further significant progresses of science. This was also a lesson bestowed by Prof. Raymond Wegmann where he created innovative research field for biology, molecular biology and biochemistry-biophysics. Progresses of developmental biology were boosted by molecular biology and reproductive engineering where ES cells and embryonic manipulation are necessary. There are no questions about the utility of their technologies. Reviews on their contributions with respect to the condition of genome manipulation are addressed. PMID- 12899460 TI - Trace elements and biominerals in the human bone. AB - Trace elements are movable from the bone before or even after death. S is most constant and Si is possible essential in maintenance of the back bone. PMID- 12899461 TI - Therapeutic vaccine to control stromal tumor-induced immunosuppression in human uterine cervixc cancer. AB - Cancer cells may escape immune surveillance by secreting in their microenvironment soluble factors that may locally paralyze the stromal effector immune cells. In the human uterine cervix cancer, HPV-16 E7 protein, released in the stroma, should contribute to cancer cells immune escape since this protein inhibits the cellular immune response to recall antigens or alloantigens and strongly enhances the release of immunosuppressive cytokines by APCs. This prompted us to prepare a therapeutic vaccine triggering anti-E7 neutralizing Abs to antagonize the E7-induced stromal immunosuppressive effects and allow cellular immune reaction towards cancer cells including specific CTLs, induced by conventional vaccine, to be effective. Since HPV-16 is a mucosotropic virus, this therapeutic vaccine has been prepared to generate systemic as well as mucosal immunity. PMID- 12899462 TI - The small proteoglycan biglycan is associated with thick collagen fibrils in the mouse decidua. AB - In mice, embryo implantation induces profound changes in the endometrium. These changes include redifferentiation of endometrial fibroblasts and extensive remodeling of extracellular matrix components. We have previously shown that, during this process, there is an impressive increase in the thickness of collagen fibrils present in decidualised areas that surround the implantation site, while collagen fibrils in non-decidualised areas and in the interimplantation site remain thin. In vitro and in vivo experiments have identified small leucine rich proteoglycans (SLRPs) as regulators of collagen fibrillogenesis. In a previous study, we demonstrated a difference between the pre-implantation and the post implantation expression and distribution of four SLRPs types in uterine tissues. The present study, utilising immunocytochemical electron microscopy, shows that biglycan is associated with the presence of thick collagen fibrils in decidualised regions of the endometrium and that decorin is associated exclusively with thin collagen fibrils in non-decidualised endometrial areas. These results strongly indicate that biglycan plays a role in collagen fibrillogenesis and probably participates in the determination of collagen fibril thickness in the mouse decidua. PMID- 12899463 TI - Health in exchange: teachers, doctors, and the strength of informal practices in Russia. AB - This article explores teachers' and doctors' informal medical exchange practices in the context of the transforming health care system in post-Soviet Russia. Despite the advent of a medical marketplace, most Russians have low incomes and cannot buy the goods and services the market offers. Instead, they bypass the formal market mechanisms (such as obtaining cheaper medicine through personal connections) and official procedures (such as obtaining free or cheaper health care services despite the emergence of paid services) by using their social networks. This paper uses a network perspective to investigate how doctors' and teachers' mutual relations are formed and what resources form the basis of these informal exchange practices. Drawing on structured diary data and qualitative interviews with 20 teachers, in addition to interviews with eight doctors in St. Petersburg, the study goes beyond mere statistics on health care and attempts to depict "from below" the implications health and illness have for survival in contemporary Russia. PMID- 12899464 TI - Toward a culture-bound syndrome-based insanity defense? AB - The American Psychiatric Association's recent inclusion of a Glossary of Culture Bound Syndromes within DSM-IV draws upon decades of medical anthropological and cultural psychiatric research to afford culture-bound syndromes (CBSs) a newfound legitimacy within professional Western psychiatric nosology. While DSM-IV's recognition of the CBS concept as a category of psychosocial distress has important clinical implications for mental health care practitioners throughout the world, it also has significant legal implications. Given that several CBSs involve a degree of psychological impairment that may satisfy the standard for legal insanity under certain circumstances, this essay focuses on the potential emergence of an insanity defense based on the claim that an immigrant or minority defendant was suffering from a CBS at the time of his or her criminal act. Aimed at initiating interdisciplinary debate over the reification of the CBS concept, the essay discusses the theoretical ambiguity and status of CBSs within professional Western psychiatry, describes what a CBS-based insanity defense might look like, and considers the relevant challenges facing medical anthropologists and cultural psychiatrists, on the one hand, and legal practitioners, on the other. The essay identifies a pressing need for interdisciplinary debate concerning the validity, scope, and viability of CBS based insanity defenses. PMID- 12899465 TI - Hard questions in court culture and psychiatry on trial. PMID- 12899466 TI - The importance of a pleasant process of treatment: lessons on healing from South India. AB - This paper considers the significance of the positive and negative aesthetic qualities of different therapies--in other words, how "pleasant" (a term that is elaborated in the paper) it is to undergo various treatments. Interviews were conducted with patients undergoing three forms of healing for mental illness and related problems in the state of Kerala in southern India--ayurvedic (indigenous) psychiatry, allopathic (biomedical) psychiatry, and religious healing. Informants revealed concerns about the aesthetic process of therapy, reporting adverse reactions to allopathic treatments and in some cases asserting that they enjoyed ayurvedic procedures. Some informants with long-term illnesses had chosen to live in the process of therapy and reside indefinitely in the aesthetically engaging environment of a mosque, temple, or church after pursuing medical therapies for years. Thus considerations of the quality of the process of therapy also call for an examination of the limitations of the concept of "cure" for describing what is accomplished in healing in some therapeutic settings. PMID- 12899467 TI - Identity, activity, and the well-being of adolescents and youths: lessons from young people in a Micronesian society. AB - Comparative studies of the health, well-being, and social functioning of adolescents and youths have, in the past two decades, been focused on young people in developing countries where rapid social, cultural, and economic changes have been associated with dramatic increases in the rates of social problems indicative of poorer mental health. Young people in many Pacific Island societies suffer from some of the highest rates of social problems like suicide and substance abuse in the world. It is generally agreed that the increases in rates of social problems among youths in this region result from increases in psychosocial stress, anger, and frustration surrounding intergenerational conflict within the family. Much less is known about the aspects of everyday experiences of young people in the Pacific that can lead to psychosocial stress and the angry episodes of interpersonal conflict that often precede suicide attempts and binge drinking. This paper examines 40 cases of interpersonal conflict in young men's and women's experiences in the islands of Chuuk of the Federated States of Micronesia to better understand what can lead to elevated levels of psychosocial stress for youths in the Pacific. This study shows that the emotional crises of young people in Chuuk often emerge from the incongruence in their pursuit of valued personal and social identities within the family, the community, and the peer group. Thus youths who experience more incongruity in their engagements across the multiple activity settings of everyday life are at greater risk for stressful experiences. PMID- 12899468 TI - Reformulation of diagnosis with attention to cultural dynamics: case of a Japanese woman hospitalized in Melbourne, Australia. PMID- 12899469 TI - Image quality in myopic eyes corrected with laser in situ keratomileusis and phakic intraocular lens. AB - PURPOSE: To compare image quality due to higher-order aberrations following laser in situ keratomileusis (LASIK) or implantation of phakic intraocular lens (PIOL) to correct high myopia. METHODS: Postoperative wavefront examinations, normalized to a pupil size of 5.5 mm, were obtained for 19 LASIK and 20 PIOL eyes for the same surgeon over the same time period. Higher-order aberrations and simulated retinal images were compared. RESULTS: For this small sample, the LASIK eyes yielded an average three times more spherical aberration and two times more coma than PIOL eyes. The effects of these differences were visualized using the simulated retinal images. CONCLUSION: Spherical aberration and coma are the major differences between postoperative LASIK and PIOL higher-order aberrations, and simulated retinal images can be used to visualize these effects. PMID- 12899470 TI - Wavefront-supported photorefractive keratectomy: 12-month follow-up. AB - PURPOSE: To evaluate safety, efficacy, predictability, and stability of wavefront supported photorefractive keratectomy (PRK) for correction of myopia and myopic astigmatism, with 12-month follow-up. METHODS: Thirty eyes of 23 patients with myopia less than -8.00 D (mean -3.76 +/- 1.90 D) and cylinder less than -3.50 D (mean -0.81 +/- 0.71 D) were selected. Aberrometry measurements were taken with the Asclepion aberrometer in order to perform customized wavefront-supported PRK. Eyes were treated with the Asclepion MEL 70 excimer laser and were followed for 12 months. RESULTS: UCVA of 20/16 or better was achieved by 47% (14 eyes) at 1 month, 67% (20 eyes) at 3 months, 77% (23 eyes) at 6 months, 90% (27 eyes) at 9 months, and 83% (25 eyes) after 1 year. No eye lost more than 1 line of BSCVA at 3, 6, 9, or 12 months. Two eyes (7%) gained more than 2 lines 1 month postoperatively, and 13% (4 eyes) gained more than 2 lines at 6, 9, and 12 months postoperatively. BSCVA of 20/10 or better was achieved in nine eyes (30%) at 1 and 12 months. Visual acuity under low contrast at 3 and 12 months after PRK was unchanged in 87% (26 eyes). Visual acuity under glare remained unchanged at 3 months after PRK in 86% (26 eyes) and at 12 months in 83% (25 eyes). CONCLUSION: Excimer laser wavefront-supported PRK with the Asclepion MEL 70 laser was safe and effective for the treatment of myopia and myopic astigmatism. Daylight visual acuity and mesopic visual acuity outcomes remained stable over 1 year. PMID- 12899471 TI - Laser epithelial keratomileusis: outcome of initial cases performed by an experienced surgeon. AB - PURPOSE: To evaluate refractive outcome and objective clinical data, and determine efficacy, predictability, and safety of laser epithelial keratomileusis (LASEK) for myopic treatments. METHODS: We performed a retrospective non comparative single-surgeon case series on the first 20 LASEK procedures (Alcon LADARVision 4000 laser). Mean patient age was 41.2 years (range 21 to 60 yr): 13 men and 7 women. Mean preoperative spherical equivalent refraction was -6.47+/ 2.78 D. Corneal haze, uncorrected and spectacle-corrected visual acuity and manifest refraction were evaluated. RESULTS: Of 20 eyes studied, 3 were corrected for monovision. In the non-monovision group, 20/40 or better visual acuity was achieved in 94% (16 eyes) at 1 month, 100% (13 eyes) at 3 months, and 91% (10 eyes) at 6 months after LASEK; 20/20 or better was achieved in 12% (2 eyes) at 1 month, 46% (6 eyes) at 3 months, and 45% (5 eyes) at 6 months. Corneal haze at 1 month was grade 0.5 in 35% (7 eyes), 1 in 20% (4 eyes) and 2 in 10% (2 eyes). At 3 months, 62% (12 eyes) had grade 0.5 and 31% (6 eyes) had grade 1. At 6 months, 58% (12 eyes) had grade 0.5, 25% (5 eyes) had grade 1, and 8% (2 eyes) had grade 2. CONCLUSIONS: LASEK is a challenging procedure. Creating the epithelial flap is not simple and may have contributed to the high haze incidence in our study. PMID- 12899472 TI - Laser in situ keratomileusis for correction of induced astigmatism from cataract surgery. AB - PURPOSE: To evaluate the efficacy, predictability, stability, and safety of laser in situ keratomileusis (LASIK) to correct residual astigmatism after cataract surgery. METHODS: LASIK was performed on 20 eyes of 20 patients with refractive myopic or mixed astigmatism (3.00 to 6.00 D) at least 1 year after extracapsular cataract extraction with posterior chamber intraocular lens implantation without complication. Each eye received bitoric LASIK with the Nidek EC-5000 excimer laser and the Automated Corneal Shaper microkeratome. RESULTS: At 6 months after LASIK, mean refractive cylinder decreased from 4.64+/-0.63 D to 0.44+/-0.24 D (P<.001). Mean percent reduction of astigmatism was 90.4+/-5.0% (range 80% to 100%). Mean spherical equivalent refraction decreased from -2.19+/-0.88 D (range 1.00 to -3.88 D) to -0.32+/-0.34 D (range -1.25 to +0.38 D) (P<.001). Vector analysis showed that the mean amount of axis deviation was 0.7+/-1.2 degrees (range 0 degrees to 4.3 degrees) and the mean percent correction of preoperative astigmatism was 92.1+/-5.9% (range 85.6% to 108%). Eighty-five percent of all eyes had a mean spherical equivalent refraction and mean cylinder within +/-0.50 D of emmetropia. Change in spherical equivalent refraction and cylinder from 2 weeks to 6 months was < or = 0.50 D in 90% (18 eyes) and 95% (19 eyes), respectively. Spectacle-corrected visual acuity was not reduced in any eye. Diffuse lamellar keratitis occurred in three eyes (15%) after LASIK, and were treated successfully with eyedrops. CONCLUSION: LASIK was an effective, predictable, stable, and safe procedure for correction of residual myopic or mixed astigmatism ranging from 3.00 to 6.00 D with a low spherical component after cataract surgery. PMID- 12899473 TI - Conductive keratoplasty to correct hyperopic astigmatism. AB - PURPOSE: To evaluate the efficacy of conductive keratoplasty in the treatment of pre-existing and surgically induced hyperopic astigmatism. METHODS: In this prospective, noncomparative case series, four eyes of four subjects, two female and two male (age 25 to 47 yr) were treated for hyperopia (up to +5.50 D) and hyperopic astigmatism (up to +5.75 D) with the Refractec ViewPoint conductive keratoplasty system. The follow-up period was 6 months. Uncorrected and spectacle corrected visual acuity, manifest and cycloplegic refraction, and videokeratographs were obtained before and after surgery. We treated two patients who had already had LASIK, one of them with a decentered ablation and the other with flap striae, one patient after PRK, and one patient with keratoconus. RESULTS: No complications were observed. No eye lost lines of spectacle-corrected visual acuity. All eyes showed improvement of uncorrected visual acuity of 3 or more lines. Videokeratographs demonstrated improved centration and reduction in keratometric power readings. Each eye was analyzed separately, including a comparative analysis of the proposed nomograms and quality of vision after surgery. CONCLUSIONS: Conductive keratoplasty may be a minimally invasive solution for patients with irregular hyperopic astigmatism, offering improved quality of vision in instances of flap striae by tightening the central cornea. PMID- 12899474 TI - Optic nerve head blood flow using scanning laser Doppler flowmetry after laser in situ keratomileusis. AB - PURPOSE: To investigate ocular blood flow changes following laser in situ keratomileusis (LASIK) in myopic eyes. METHODS: Seventeen eyes of ten myopic patients were included in the study. Mean age of the seven men and three women was 29.1+/-2.5 years. Mean spherical equivalent refraction was -7.70+/-2.52 D. LASIK was performed with the Hansatome microkeratome and Summit SVS Apex excimer laser. Optic nerve head blood flow was measured by using the Heidelberg Retinal Flowmeter preoperatively and postoperatively at 1 day, 1 week, and 1 month. Preoperative ocular blood flow data (blood flow, volume, velocity) were compared with postoperative data using variance analysis. Correlation analysis was performed for age of patients, suction time, and ocular blood flow data. RESULTS: Mean suction time was 19+/-7 seconds. There was no significant correlation between suction time, age, and ocular blood flow. We found an increase in blood flow at the lamina cribrosa taken 1 day postoperatively (P=.04). Blood flow returned to preoperative values at the 1-week and 1-month measurements in the lamina cribrosa region. There was no significant change in ocular blood flow data in the neuroretinal rim except an increase in velocity at the lower nasal quadrant on postoperative day 1 (P=.03). CONCLUSIONS: LASIK caused a temporary increase in blood flow at the lamina cribrosa region of the optical nerve head in healthy myopic eyes. The increase in blood flow may suggest a compensatory change for maintaining blood flow in the ocular nerve head following LASIK-induced ischemia. PMID- 12899475 TI - Technical improvements in photorefractive keratectomy for correction of high myopia. AB - PURPOSE: To evaluate the effects of hardware and software improvements in photorefractive keratectomy (PRK) for the treatment of highly myopic eyes. METHODS: A retrospective study was carried out in 554 patients (582 eyes) with myopia between -7.00 and -17.00 D (mean -11.20 +/- 3.60 D) who had undergone PRK using the Aesculap Meditec laser. Group 1 with a 5-mm-diameter single ablation zone; Group 2 with a 5-mm-diameter single zone surrounded by a 2-mm tapered transitional zone; Group 3 and Group 4, same as Group 2 but with the laser upgraded with a smoke aspiration control system (Group 3), and with a computer controlled fluence (Group 4). RESULTS: In Group 1 at 2 years after PRK (50 eyes), 10 eyes (20%) were within +/-1.00 D of attempted correction and 13 eyes (26%) were within +/-2.00 D. In Group 2 at 2 years (118 eyes), 42 eyes (36%) were within +/-1.00 D of attempted correction and 65 eyes (55%) were within +/-2.00 D. In Group 3 at 2 years (43 eyes), 18 eyes (42%) were within +/-1.00 D of attempted correction and 28 eyes (65%) were within +/-2.00 D. In Group 4 at 2 years (47 eyes), 25 eyes (53%) were within +/-1.00 D of attempted correction and 29 eyes (62%) were within +/-2.00 D. CONCLUSIONS: Software and hardware improvements facilitated PRK correction of high myopia with reasonable predictability, especially if a tapered transition zone was used. At 12 and 24-month follow-up, only the use of a tapered transition zone was associated with a statistically significant improvement in predictability. PMID- 12899476 TI - Influence of ablation parameters on refractive changes after phototherapeutic keratectomy. AB - PURPOSE: The aim of the current study was to control the hypothetical effects of decreased laser energy delivered to the peripheral cornea during phototherapeutic keratectomy (PTK) and provide quantitative calculation of induced low and high order aberrations. METHODS: We employed a model eye to simulate the refractive effect of homogeneous laser corneal irradiation, as in PTK, for different laser fluences (range 125 to 225 mJ/cm2) and treatment depths up to 200 microm. RESULTS: The hyperopic shift induced by the relatively lower energy delivered at the peripheral ablation zone during PTK was proportional to the treatment depth and inversely proportional to the energy fluence. The hyperopic shift calculated using the above ablation parameters was lower compared to previously reported clinical results. Higher order wavefront aberration (total root mean square) changes were of minimal significance for treatment depths up to 200 microm. CONCLUSIONS: After PTK, a hyperopic shift cannot be attributed to the energy delivery method alone. Modification of laser energy delivery algorithms may only minimize PTK-induced hyperopia. PMID- 12899477 TI - Scraping and mitomycin C to treat haze and regression after photorefractive keratectomy for myopia. AB - PURPOSE: To evaluate the efficacy, safety, and predictability of therapeutic scraping and application of a diluted 0.02% mitomycin C solution to treat haze and regression after photorefractive keratectomy (PRK) for myopia. METHODS: We performed a non-comparative, non-randomized retrospective study of 35 eyes of 30 patients who had previously undergone PRK for myopia and developed haze and regression after treatment. The range of refractive error after regression was 0.75 to -5.50 D (mean -2.92 D). Haze, ranging from grade 3 to 4 (scale: 0 to 4) caused a best spectacle-corrected visual acuity loss of 1 to 6 Snellen lines. All eyes were treated 6 to 12 months after PRK by scraping the stromal surface and application of a 0.02% mitomycin C solution for 2 minutes using a soaked merocel sponge. No laser ablation was performed. Corneal transparency, refractive error, and visual outcomes were evaluated over a 12-month follow-up period. RESULTS: All eyes had significant improvement in corneal transparency. Thirty-one eyes had haze grades lower than 1, which were maintained over time. Only four eyes showed haze grades 1 to 2 and two of them needed further treatment using the same technique. At 12 months after treatment, all eyes were within +/-1.50 of original intended correction (mean 0.04 +/- 0.63 D). BSCVA improved in all eyes. No toxic effects were observed during re-epithelialization or during follow-up. CONCLUSION: Single application of diluted mitomycin C 0.02% solution following scraping of the corneal surface was effective and safe in treating haze and regression after PRK. PMID- 12899478 TI - Interface fluid after laser in situ keratomileusis. AB - PURPOSE: To describe a case of interface fluid after laser in situ keratomileusis (LASIK) and review the literature on this rare complication after LASIK. METHODS: We present a case report and literature review. Articles for this review were chosen from electronic database and manual literature searches. MEDLINE searches were made from 1990 to April 2002, using the key words "interface fluid" and "LASIK." RESULTS: A 40-year-old man had uneventful LASIK for residual refractive error from previous penetrating keratoplasty in his right eye. Diffuse lamellar keratitis began 1 day postoperatively. Topical corticosteroids were administered. Six weeks after LASIK, a layer of interface fluid developed. Intraocular pressure was 9 mmHg when measured centrally by Goldmann applanation tonometry and 30 mmHg by Tono-pen tonometry. The interface fluid resolved with antiglaucoma agents and corticosteroids combined with cyclosporine. CONCLUSIONS: This case, along with other reported cases, demonstrate the clinical features of interface fluid after LASIK. PMID- 12899479 TI - Rupture of radial keratotomy incisions by blunt trauma 6 years after combined photorefractive keratectomy/radial keratotomy. AB - PURPOSE: To report a case of traumatic corneal rupture that occurred 6 years previous, following combined radial keratotomy (RK) and photorefractive keratectomy (PRK). METHODS: A 28-year-old man experienced a severe direct trauma to the right eye. Upon initial examination, a ruptured globe was diagnosed. Four of the eight radial incisions were ruptured with extrusion of intraocular tissues. The patient was also diagnosed with a fracture of the medial wall of the right orbit. The patient received immediate surgery during which the radial incisions that had been torn were stitched with a suture. RESULTS: Following the postoperative period, the patient had visual acuity of light perception. CONCLUSION: PRK, carried out on a cornea already treated with RK, may increase the likelihood of rupture in blunt trauma. PMID- 12899480 TI - Choroidal neovascularization after LASIK. PMID- 12899481 TI - Decentration of the optical zone: centering on the pupil versus the coaxially sighted corneal light reflex in LASIK for hyperopia. PMID- 12899482 TI - Treatment of presbyopia by infrared laser radial sclerectomy. PMID- 12899483 TI - Spatial vision under low luminance after laser refractive surgery. PMID- 12899484 TI - What's new in first trimester ultrasound. AB - There are several advantages to ultrasound examination in early pregnancy. Ultrasound performed during the first trimester confirms an intrauterine pregnancy, establishes accurate dating, and is crucial in diagnosing early pregnancy failure and ectopic pregnancy. As sonographic spatial resolution continues to improve, first trimester sonography increasingly will offer early pregnancy screening for chromosomal abnormalities and fetal structural abnormalities. PMID- 12899485 TI - Ultrasound detection of first trimester malformations: a pictorial essay. AB - Knowledge of normal and abnormal anatomy in the first trimester aids in early detection of anomalies and the avoidance of potential pitfalls. PMID- 12899486 TI - Prenatal diagnosis for detection of aneuploidy: the options. AB - The value of all noninvasive prenatal tests must be viewed with the perspective of the consequences of invasive testing. Regarding second trimester noninvasive testing, biochemical screening is more accurate in establishing risk than maternal age alone. One or more major ultrasound abnormalities, nuchal thickening, or a shortened humerus should raise concern for Down syndrome regardless of the patient's a priori risk based on age or biochemical markers. Isolated minor ultrasound markers should not be used in calculating risk in low risk patients regarding Down syndrome unless the biochemical profile already places the patient at risk or in a borderline risk zone. If the ultrasound finding is hyperechoic bowel, problems other than aneuploidy may be the cause, including cystic fibrosis, infection, or hemorrhage, and these problems must be considered if hyperechoic bowel is an isolated finding. Improved risk adjustment seems to be applicable to a priori high-risk patients with completely normal sonograms. Genetic sonograms with specific risk adjustment schemata may be used to adjust a priori risk (either maternal age or biochemical screening results) at centers in which this has proven to be accurate, but whether this is statistically sound remains to be determined. The goal of second trimester ultrasound screening is to identify at-risk fetuses better and offer invasive testing to a more select group of patients. As the value of first trimester screening becomes more evident and practical, and if the risk of chorionic villus sampling becomes an acceptable norm, the patient population that reaches the second trimester of pregnancy will be select. Therefore, we can anticipate that second trimester screening and invasive testing may be needed only in a minority of cases, and the practice standards of prenatal testing and sonography (including minor ultrasound markers) will change entirely. PMID- 12899487 TI - Complications of monochorionic twins. AB - Sonography has made a dramatic impact on the obstetric management of complicated twin pregnancies. This is based in part on the ability to use prenatal US to diagnose syndromes and complications of MC twinning. All twin pregnancies are at high risk for perinatal morbidity and mortality compared with singleton gestations, but when one of the described complications is recognized, the difficulties in management are compounded dramatically. Despite the relative rarity of some of the entities described, it is vitally important to be familiar with these problems and their sonographic evaluation and diagnosis. PMID- 12899488 TI - Tips and tricks of fetal MR imaging. AB - The technique of performing fetal MR examinations differs from routine pelvic MR imaging because the fetus is in motion, and image planes must be selected orthogonal to fetal anatomy. The small size of the fetus also increases examination difficulty because thin slices (limited by SNR) are often needed for adequate examination of anatomy and pathology. Knowledge of fetal anatomy and neonatal anatomy/ pathology are needed in the assessment of fetal MR imaging. Knowledge of MR artifacts aids in image interpretation. PMID- 12899489 TI - MR imaging of pelvic floor relaxation. AB - The wide variety of available surface coils, pulse sequences, and post-processing techniques make MR imaging a useful clinical and research tool for evaluation of pelvic floor relaxation. Cases of isolated cystocele do not require imaging; however, in cases in which multiple compartments of the pelvis are involved or the patient has failed prior surgery, MR imaging should be considered for preoperative planning. PMID- 12899490 TI - Imaging of female infertility. AB - Normal reproduction requires healthy female anatomy. Cervical, uterine, tubal, ovarian, and peritoneal factors can coexist and cause female infertility. Ultrasound, hysterosalpingography, MR imaging, and fallopian tube catheterization are the radiologists' armamentarium for diagnosis. This article illustrates important findings in the infertile woman. An understanding of these entities helps i n accurateand sympathetic treatment. PMID- 12899491 TI - Ultrasonographic evaluation of the endometrium in postmenopausal vaginal bleeding. AB - Transvaginal ultrasound with SIS is a cost-minimizing screening tool for perimenopausal and postmenopausal women with vaginal bleeding. Its use decreases the need for invasive diagnostic procedures for women without abnormalities, and ultrasound increases the sensitivity of detecting abnormalities in women with pathologic conditions. Vaginal sonography is preferred over uniform biopsy of postmenopausal women with vaginal bleeding because it (1) is a less invasive procedure, (2) is generally painless, (3) has no complications, and (4) may be more sensitive for detecting carcinoma than blind biopsy. Transvaginal sonography is rarely nondiagnostic. Endometrial sampling is less successful in women with a thin endometrial stripe on ultrasound than in women with real endometrial pathologic condition. A limitation of ultrasound is that an abnormal finding is not specific: ultrasound cannot always reliably distinguish between benign proliferation, hyperplasia, polyps, and cancer. Although ultrasound may not be able to distinguish between hyperplasia and malignancy, the next step in the clinical treatment requires tissue sampling. Because of the risk of progression of complex hyperplasia to carcinoma, patients with this finding may benefit from hormonal suppression, dilatation and curettage, endometrial ablation, or hysterectomy, depending on the clinical scenario. The inability to distinguish these two entities based on ultrasound alone should not be seen as a limitation because tissue sampling is required in either case. Occasionally (in 5% to 10% of cases), a woman's endometrium cannot be identified on ultrasound, and these women also need further evaluation. Ultrasonography also may be used as a first-line investigation in other populations with abnormal uterine bleeding. In a multicenter, randomized, controlled trial of 400 women with abnormal uterine bleeding, the investigators found that transvaginal sonography combined with Pipelle endometrial biopsy and outpatient hysteroscopy was as effective as inpatient hysteroscopy and curettage. The subject, included women older than 35 years with PMB, menorrhagia, intermenstrual bleeding, postcoital bleeding, or irregular menses. Transvaginal sonography may be a cost-effective. sensitive, and well-tolerated method to evaluate most women with abnormal bleeding in combination with physical examination and endometrial biopsy and hysteroscopy us indicated. Hysteroscopy is likely to become the new gold standard in the future because of its ability to visualize directly the endometrium and perform directed biopsies as indicated. As office-based hysteroscopy becomes more practical and widespread, the technique may become more cost effective. An evaluation plan using transvaginal sonography as the initial screening evaluation followed by endometrial biopsy or, more likely, hysteroscopy is likely to become the standard of care (Fig. 12). It remains unproven whether certain patients at higher risk for carcinoma should proceed directly to invasive evaluation. Women on tamoxifen with persistent recurrent bleeding, women with significant risk factors for carcinoma, and women with life-threatening hemorrhage comprise this group. Further studies are still necessary to evaluate high-risk patients and determine whether ultrasound or biopsy is really the most cost-effective initial test. PMID- 12899492 TI - Sonohysterography. AB - Sonohysterography can distinguish focal from diffuse pathology reliably and has become a crucial imaging test in the triage of PMB and in premenopausal patients with dysfunctional uterine bleeding or infertility. Polyps and submucosal fibroids are the most common focal findings during SHG. In postmenopausal patients, detection and accurate localization of findings, rather than lesion characterization, are the primary goals of the procedure. Most, if not all, focal lesions in this patient population require tissue diagnosis, even when the imaging features suggest benign lesions. PMID- 12899493 TI - MR imaging of the ovaries: normal appearance and benign disease. AB - MR imaging enables a physician to make an accurate diagnosis of various benign adnexal masses and helps to obviate unnecessary surgery. PMID- 12899494 TI - Osteoporosis imaging. AB - Because osteoporotic fractures may be prevented, diagnostic techniques are essential in the assessment of osteoporosis. Conventional radiographs of the spine are not suited for diagnosing early osteoporosis, but they show fractures that may have no clinical symptoms. The radiologist should be aware of the enormous significance of these fractures for future osteoporotic fractures. Bone mass measurements are standard techniques in the diagnosis of osteoporosis, which are the basis of the WHO definition of osteoporosis. In this article the authors presented these standard techniques and newer diagnostic techniques that provide insights in the structure of trabecular bone. PMID- 12899495 TI - Current uses of ultrasound in the evaluation of the breast. AB - Ultrasound is an important imaging modality in evaluating the breast. One of the most common uses of ultrasound is to help distinguish benign from malignant breast disease, primarily with gray-scale ultrasound but also with Doppler ultrasound. Another common use is to provide guidance for interventional procedures. Less common uses include assisting in staging of breast cancer and evaluating patients with implants. Recently there has been an interest in using ultrasound to screen asymptomatic women for breast cancer, as is done with mammography. Further studies must be performed to assess if this reduces mortality from breast cancer. Although primarily used to image the female breast, ultrasound also can be used to evaluate breast-related concerns in men. Uses of contrast-enhanced ultrasound are still experimental and would add an invasive component to an otherwise noninvasive study. PMID- 12899496 TI - On the long and winding road to an evidence-based diet. PMID- 12899497 TI - Colorectal cancer. AB - Colorectal cancer (CRC) is one of the leading causes of mortality and accounts for approximately 200000 deaths per year in Europe and the USA. Chemotherapy and radiotherapy have established roles in the treatment of colorectal cancer and can contribute to cure rate, prolongation of survival, reduction of local rates of recurrence and enhanced quality of life in patients with advanced disease. However, over the past few years there have been major advances in our understanding of the molecular basis of this tumour and its progression from adenoma to carcinoma that hold potential for translation into novel strategies for the treatment of CRC. Furthermore, newer agents directed against different intracellular targets have also been shown to be efficacious in CRC treatment. Such improvements should continue to lead to increased cure rates in early disease and better overall survival in advanced cases. The aim of this review is to provide an overview of the pathogenesis of CRC and to present the evidence base for chemotherapy and some of the novel strategies that are currently being evaluated in phase I and II trials. PMID- 12899498 TI - Psychosocial issues in cancer genetics--current status and future directions. AB - Diagnostic tests are now available that allow genetic testing for several types of cancer. The aim of genetic counseling and testing for cancer is to educate individuals about cancer risk and cancer prevention, which it is hoped will lead to a reduction in morbidity and mortality. However, at this relatively early stage in the development of genetic counseling and testing programs, information is needed on the psychosocial impact of such programs on both the individual counselee and his/her family. This paper reviews the findings obtained during the past decade on the uptake of genetic testing, reasons for undergoing genetic testing, and the impact of genetic counseling and testing on feelings of distress and guilt. Specific attention is paid to experiences with prophylactic mastectomy and oophorectomy and the effectiveness of the uptake of and satisfaction with these risk-reducing procedures. In addition, the possible impact of genetic testing on insurance, work and future plans is discussed. Suggestions are given for translating research findings into psychosocial services and future research efforts. PMID- 12899499 TI - Randomized trial with fruits and vegetables in prevention of cancer. AB - Along with tobacco use, diet has the greatest impact on the development of human cancer. Within the broad category 'diet', the consumption of fruits and vegetables apparently plays a dominant role. Considerable efforts have been made to prove the preventive effect of different kinds of fruits and vegetables but randomized chemoprevention trials have failed to prove this presumed effectiveness of their single ingredients. The conclusive demonstration of a cancer-protective effect of a high consumption of fruits and vegetables is considered to be impractical. However, current historical changes in Europe offer a unique opportunity to conduct such a randomized trial in specific European countries. This study describes the nutritional situation and the conditions of the health system in the Baltic countries as appropriate geographic areas and demonstrates some basic design issues of the trial for three variants of outcome assumptions. A realistic assumption would be that a trial needs about 30000 participants, an intervention period of 10 years and a subsequent follow-up time of 20 years. Annual costs could range between dollars 5 and dollars 10 million. A high intake of fruits and vegetables should be proven scientifically as a valid tool for cancer prevention. For a comparably short period the Baltic countries offer a time-window for a randomized trial. It is unlikely that the costs of such a trial would considerably exceed the costs of the available chemoprevention trials. PMID- 12899500 TI - Chemotherapy and antiangiogenesis--drug-specific, dose-related effects. AB - Dose-response effects of fluorouracil, paclitaxel, doxorubicin, cisplatin, methotrexate, cyclophosphamide and etoposide on VEGF165/164-mediated angiogenesis using the rat mesenteric-window angiogenesis assay are reported. VEGF is a pivotal pro-angiogenic factor in most tumors. Microvessel spatial extension, density, pattern formation and segment length were assessed quantitatively and objectively. A single i.v. injection of each drug was given at a low, intermediate or high dose, 7 days before sacrifice. All the drugs elicited significant responses in terms of one or more measured variables. Only paclitaxel, doxorubicin and cyclophosphamide significantly suppressed the overall angiogenic response (p < or = 0.0001, p < or = 0.0002 and p < or = 0.05, respectively), however. Taking toxicity into account, paclitaxel was more potent in inhibition of angiogenesis than the other agents. No clear correlation was found between drug half-life, the degree of toxic effects (in terms ofbody weight changes) and the antiangiogenic effect. The antiangiogenic effects were distinctly drug specific. PMID- 12899501 TI - Effects of probucol on endothelial damage by 5-fluorouracil. AB - Cardiotoxicity is a serious side effect of cancer treatment with the commonly used drug 5-fluorouracil (5-FU). The pathophysiology of this is unclear. Experimental studies show a thrombogenic effect of 5-FU, secondary to a direct toxic effect on the endothelium, possibly mediated by radical generation. Probucol is a lipid-lowering drug with strong antioxidant properties. The aim of this study was to evaluate the possibility of using probucol treatment to protect against the toxicity of 5-FU on vascular endothelium of the central artery in the ears of rabbits. Five groups of rabbits were treated with 1) 5-FU, 2) saline, 3) probucol high-dose and saline, 4) probucol high-dose and 5-FU, 5) probucol low dose and 5-FU. Damage to the arterial endothelium was evaluated by scanning electron microscopy. Damage to the endothelium in 5-FU + probucol-treated animals was minimal and comparable to that of the control group. Intima disruption or thrombus formation was seen with 5-FU only. The results of the study indicate that treatment with probucol prevents 5-FU-induced endothelial injury. PMID- 12899502 TI - Vinorelbine, epirubicin and fluorouracil as first-line therapy in metastatic breast cancer--a phase II trial. AB - A phase II study was conducted to assess the toxicity and response rate of vinorelbine (NavelbineR) combined with epirubicin and fluorouracil (NEF) in metastatic breast cancer. Vinorelbine was delivered at a dose of 25 mg/m2 on days 1 and 8, epirubicin at 60 mg/m2 on day 1 and fluorouracil at 600 mg/m2 on day 1, at 3-week intervals. Forty consecutive ambulant patients with breast cancer with measurable metastases were treated with a total of 310 cycles (median 8) as first line therapy. The objective response rate was 83% (95% CI 71-95) (6/40 CR 15%, 27140 PR 68%). In 3 patients, CNS metastases were detected during NEF therapy those who had a partial response in their visceral metastases. Median time to progression was 13 months (95% CI 7-19) and estimated median survival time was 32 months. The main dose-limiting adverse effect, grade III-IV haematological toxicity, was reported in 92% of patients. One patient died of neutropenic sepsis. Grade III infections requiring hospitalization were observed in 8 patients (20%). Half of the patients complained of mild constipation, nausea or stomatitis, which were easily managed. Almost all patients had grade III alopecia. One patient with previous adjuvant anthracycline therapy (CEF x 9 two years earlier) developed fatal grade IV cardiac failure associated with pulmonary emboli 2 months after completion of NEF therapy (PR with 6 cycles). In line with the observations of others conducting phase II first-line trials combining vinorelbine and epirubicin, it is concluded that the NEF regimen is effective in metastatic breast cancer. Haematological toxicity, however, requires dose reductions in many patients. Furthermore, careful monitoring of cardiac function is necessary, particularly in patients who received prior adjuvant anthracycline therapy. PMID- 12899503 TI - Cisplatin and radiotherapy in the treatment of locally advanced head and neck cancer--a review of their cooperation. AB - A review of the published literature has been undertaken to ascertain the trends in treatment schedules of unresectable head and neck cancer (HNC) using cisplatin in conjunction with radiotherapy. In addition, four trials were reviewed where cisplatin was examined as a single agent without radiotherapy, demonstrating that cisplatin alone has a palliative effect only, and for a curative intent should be used in combination with radiation. Of the numerous clinical trials published on such combined chemo-radiotherapy, 16 were selected for analysis fulfilling the following criteria: cisplatin used as the sole chemotherapeutic agent in combination with radiation for patients with unresectable squamous cell carcinomas of the head and neck, with no previous treatment for the same malignancy. Daily low-dose cisplatin performed in 6 out of the 16 trials demonstrated increased tumour control with less toxicity as compared to weekly high-dose drug delivery. The increased tumour control with a daily low-dose schedule reflects the observation that cisplatin dissociates from the DNA in 24 h. Other cisplatin properties (e.g. radiosensitizing) also have the potential to influence chemo-radiotherapy treatment outcomes. There is significant scope to further optimize the treatment schedule for the unresectable HNC through detailed study of the pharmacokinetics and radiobiology of combined chemo-radiotherapy. PMID- 12899504 TI - Daily-diary evaluated side effects of dose-escalation radiotherapy of prostate cancer using the stereotactic BeamCath technique. AB - The aim of this study was to evaluate self-assessed late side effects in patients with prostate cancer treated with frameless stereotactic dose-escalation radiotherapy using BeamCath, a new technique that has been developed for accurate positioning of the prostate at treatment set-up, and in which a specially designed urethral catheter containing high-density fiducial markers is used. The method was tested in the first 104 patients in a Scandinavian dose-escalation study with doses up to 76 Gy. Side effects were reported in a daily diary and evaluated at the start of treatment (baseline) and at 1-year follow-up. The patients were compared with those treated with conventional (n = 53) and conformal techniques (n = 175). Dose-escalation radiotherapy (76 Gy) decreased urinary frequency, urgency and starting problems at 1-year in comparison with baseline. The dose-escalation therapy did not induce any increase in gastrointestinal side effects in comparison with the effect of conformal therapy < or = 70 Gy at the 1-year follow-up, apart from a slight increase in rectal mucus in the 76 Gy group. All groups, except patients receiving the 74 Gy with smaller fields, reported a slight increase in gastrointestinal toxicity at 1-year compared with baseline. Dose-escalation radiotherapy of prostate cancer using the BeamCath technique did not induce any significant increase in late side effects in comparison with conformal technique. PMID- 12899505 TI - Clinico-pathological features of thyroid cancer as observed in five referral hospitals in Iran--a review of 1177 cases. AB - In this study some of the clinical and pathological aspects of thyroid carcinomas in Iran are defined. Until fairly recently, Iran was an area of endemic iodine deficiency. The medical records of 1177 confirmed cases of thyroid cancer treated over a 15-year period at 5 tertiary referral centers were reviewed. The female/male ratio was 1.8/1. Mean age was 42.8 +/- 0.9 years, male patients being significantly older. The most common clinical presentation (initial manifestation) was a central neck mass; 28.6% of the tumors had metastasized, usually to the cervical lymph nodes, by the time the patient presented with disease. The mean size of non-metastatic tumors was 5.0 cm at their longest diameter. The distribution of tumors in our study did not reflect the expectations for an iodine-deficient area, where follicular thyroid carcinoma is common, but rather what is seen in iodine-rich areas. Papillary and follicular types accounted for79.7% and 8.8% of cases, respectively. PMID- 12899506 TI - Prognostic factors for the survival of surgically treated patients for non-small cell lung cancer. AB - The survival and outcome rates of 284 patients who underwent surgical treatment for non-small cell lung cancer were assessed retrospectively. Resectability rate was 94.1%, hospital mortality 3.9% (n = 11) and the mortality rates in patients who underwent pneumonectomy or lobectomy were 8.9% and 0.6%, respectively. The overall 5-year survival was 43.6%. Female gender, earlier stages of disease and a complete resection were strongly predictive for a long-term survival. Women in stage IA disease had a 5-year survival rate of 92.7%. The 5-year survival rate for patients in stages IIIA and N2 disease who underwent a complete resection was 21.9%, and 9% for those who did not undergo a complete resection. It is concluded that the best surgical results were observed in women who were operated on at an early stage of disease. A complete resection also contributed to a better outcome, even for patients in stage IIIA and N2 disease. PMID- 12899507 TI - Cardiac toxicity associated with capecitabine therapy. PMID- 12899508 TI - Epidermal growth factor receptor inhibition induces trichomegaly. PMID- 12899509 TI - Gastrointestinal cancer and the long-term use of pravastatin in the elderly. PMID- 12899510 TI - Technetium-99m-tetrofosmin myocardial imaging in cardiotoxicity screening for left-sided breast cancer irradiation. PMID- 12899511 TI - Evolution and function of vitamin D. AB - It is remarkable that phytoplankton and zooplankton have been producing vitamin D for more than 500 million years. The role of vitamin D in lower non-vertebrate life forms is not well understood. However, it is critically important that most vertebrates obtain an adequate source of vitamin D, either from exposure to sunlight or from their diet, in order to develop and maintain a healthy mineralized skeleton. Vitamin D deficiency is an unrecognized epidemic in most adults who are not exposed to adequate sunlight. This can precipitate and exacerbate osteoporosis and cause the painful bone disease osteomalacia. Once vitamin D is absorbed from the diet or made in the skin by the action of sunlight, it is metabolized in the liver to 25-hydroxyvitamin D [25(OH)D] and then in the kidney to 1,25-dihydroxyvitamin D [1,25(OH)2D]. 1,25(OH)2D interacts with its nuclear receptor (VDR) in the intestine and bone in order to maintain calcium homeostasis. The VDR is also present in a wide variety of other tissues. 1,25(OH)2D interacts with these receptors to have a multitude of important physiological effects. In addition, it is now recognized that many tissues, including colon, breast and prostate, have the enzymatic machinery to produce 1,25(OH)2D. The insights into the new biological functions of 1,25(OH)2D in regulating cell growth, modulating the immune system and modulating the renin angiotensin system provides an explanation for why diminished sun exposure at higher latitudes is associated with increased risk of dying of many common cancers, developing type 1 diabetes and multiple sclerosis, and having a higher incidence of hypertension. Another calciotropic hormone that is also produced in the skin, parathyroid hormone-related peptide, is also a potent inhibitor of squamous cell proliferation. The use of agonists and antagonists for PTHrP has important clinical applications for the prevention and treatment of skin diseases and disorders of hair growth. PMID- 12899512 TI - Current understanding of the function of the nuclear vitamin D receptor in response to its natural and synthetic ligands. AB - The vitamin D receptor (VDR), the high affinity receptor for 1alpha,25 dihydroxyvitamin D3 (1alpha,25(OH)2D3), is a member of the nuclear receptor superfamily. VDR preferentially forms a heterodimeric complex with the retinoid X receptor (RXR) and binds to 1alpha,25(OH)2D3 response elements (VDREs) that consist of two hexameric motifs in a directly repeated (DR) or inverted palindromic (IP) arrangement. DNA-complexed VDR acts as a molecular switch of nuclear 1alpha,25(OH)2D3 signaling by transmitting its activation status to different chromatin loci containing the 1alpha,25(OH)2D3 target genes. Approximately 0.5% of the human genome (about 200 genes) are estimated to be primary targets of 1alpha,25(OH)2D3, but via various mechanisms the VDR appears to interfere in the regulation of even more genes. The molecular basis of the regulatory actions of 1alpha,25(OH)2D3 and its synthetic analogs are ligand triggered protein-protein interactions of the ligand-binding domain (LBD) of the VDR with coactivator (CoA), corepressor (CoR) and other nuclear proteins. Most analogs have been identified as agonists, a few as antagonists (ZK159222 and TEI 9647) and only Gemini and some of its variations as nonagonists. The positioning of helix 12 of the LBD is of critical importance for the agonistic, antagonistic and nonagonistic conformation of the VDR. In each of the three conformations, the VDR performs different protein-protein interactions, which then result in a characteristic functional profile. The functional profile of some 1alpha,25(OH)2D3 analogs, such as EB1089 and Gemini, can be modulated by protein and DNA interaction partners of the VDR. This provides them with some selectivity for DNA-dependent and -independent signaling pathways and VDRE structures. PMID- 12899513 TI - Approaches to evaluating the association of vitamin D receptor gene polymorphisms with breast cancer risk. AB - The steroid hormone 1,25 dihydroxyvitamin D3 is thought to protect against breast cancer. Its actions are mediated via the vitamin D receptor (VDR) and a number of polymorphisms in the VDR gene have been identified, some of which may alter susceptibility to breast cancer. This study has investigated whether specific VDR gene polymorphisms are associated with breast cancer risk in a UK Caucasian population. Female breast cancer patients (n = 313) and control women with a negative screening mammogram (n = 410) were recruited and their VDR polymorphisms were determined. The 3' VDR polymorphism BsmI was significantly associated with breast cancer risk; odds ratio bb vs. BB genotype = 1.79 (95% CI, 1.12-2.86; P = 0.0221). In addition, over 70% of seven commonly used breast cancer cell lines were found to have the at-risk genotype bb. The 5' FokI gene variant was not associated with breast cancer risk. Further investigations into how these different genotypes may affect the functional mechanisms of the VDR will provide a better strategy for identifying women at risk of breast cancer and for developing improved treatments. PMID- 12899514 TI - Ligand structure-function relationships in the vitamin D endocrine system from the perspective of drug development (including cancer treatment). AB - It has become readily apparent to many scientists and pharmaceutical companies that the vitamin D endocrine system offers a wide array of drug development opportunities. There are already successes, as noted by 1alpha,25(OH)2D3 (Roche, and Abbott) for renal osteodystrophy and osteoporosis and 1alpha(OH)D3 (Leo, Chugai, Teijin) for renal osteodystrophy and (in Japan) osteoporosis, 1alpha,24(OH)2-24-cyclopropyl-D3 (Dovonex) and 1alpha,24(OH)2D3 (Teijin) for psoriasis, and 19-nor-1alpha,25(OH)2D2 (Abbott) for renal osteodystrophy, as well as drugs under active development. Yet there are still many important and challenging drug development frontiers, particularly in the area of cancer treatment and immune system disorders where exploration is only in the initial early stages. In addition, the application of vitamin D-related drugs in neurology and brain pathology should not be overlooked. It is to be hoped that the cellular and molecular basis for the vexing problem of analog-induced hypercalcemia will be elucidated. Given that there are believed to be over 2000 analogs of 1alpha,25(OH)2D3 already available for consideration, it is to be expected that over the next decade a significant number of new vitamin D structure-function drug development projects will be brought to conclusion. PMID- 12899515 TI - Antiproliferative signalling by 1,25(OH)2D3 in prostate and breast cancer is suppressed by a mechanism involving histone deacetylation. AB - Breast and prostate cancer are leading causes of cancer death in the Western world. Hormone ablation is the primary therapy for invasive disease, but the tumour often recurs in an androgen or oestrogen receptor negative form for which novel therapies are sought urgently. The vitamin D receptor (VDR) may provide an important alternative therapeutic target. However, cancer cell line models from these tissues display a range of sensitivities to the antiproliferative effects of 1alpha,25dihydroxyvitamin D3 (1alpha,25(OH)2D3). The reason for apparent 1alpha,25(OH)2D3 insensitivity is currently unknown and we have investigated epigenetic mechanisms that may suppress the transcriptional activity of the VDR. Nuclear co-repressors have associated histone deacetylase (HDAC) activity, which keeps chromatin in a closed, transcriptionally silent state. We have found that the aggressive cancer cell lines with relative insensitivity to 1alpha,25(OH)2D3 have elevated nuclear co-repressor levels. For example, PC-3 prostate cancer cells have a significant 1.8-fold elevation in the co-repressor SMRT compared to normal epithelial cells (P < 0.05). We believe that a combination of elevated co repressor level with reduced VDR content can cause 1alpha,25(OH)2D3 resistance. Consistent with this, we have shown that combining a low dose of HDAC inhibitor Trichostatin A (15 nM TSA) with 1alpha,25(OH)2D3 (100 nM) synergistically inhibits the proliferation of PC-3 prostate and MDA-MB-231 breast cancer cell lines. The inhibition of proliferation was potentiated further by treating cells with 19-nor-hexafluoride vitamin D3 analogues instead of 1alpha,25(OH)2D3, plus TSA. For example, the combination of 1alpha,25(OH)2D3 and TSA-inhibited MDA-MB 231 cell proliferation by 38% (+/-5%), whereas Ro26-2198 (1alpha,25-(OH)2-16,23Z diene-26,27-F6-19-nor-D3) and TSA inhibited growth by 62% (+/-2%). Therapeutically the hypercalcaemic side effects associated with 1alpha,25(OH)2D3 could be minimized by combining low doses of potent 1a,25(OH)2D3 analogues with HDAC inhibitors as a novel anticancer regime for hormone-insensitive prostate and breast cancer. PMID- 12899516 TI - Squamous cell carcinomas fail to respond to the prodifferentiating actions of 1,25(OH)2D: why? AB - 1,25(OH)2D regulates a number of cellular events which contribute to its ability to stimulate differentiation of the keratinocyte. 1,25(OH)2D raises the intracellular calcium (Cai) level in part by increasing the expression of the calcium receptor (CaR). This sensitizes the cell to extracellular calcium, triggering the signaling pathway coupled to the CaR, which results in a rise in Cai. 1,25(OH)2D induces the family of phospholipases C (PLC). These enzymes mediate the hydrolysis of phosphatidyl inositol bisphosphate (PIP2) to form inositol tris phosphate (IP3) and diacylglycerol (DG), which stimulate calcium release from intracellular stores and activate protein kinases C (PKC), respectively. The CaR and other G protein coupled receptors signal through PLC beta, whereas tyrosine kinase growth factor receptors such as the EGF receptor signal through PLC-gamma. Calcium and PKC regulate the expression of genes in part by controlling the levels and activity of AP-1 transcription factors. 1,25(OH)2D also directly induces structural genes such as involucrin, a substrate for transglutaminase, which crosslinks it to other substrates to form the cornified envelope. 1,25(OH)2D regulates gene expression by activating the vitamin D receptor (VDR), a transcription factor, which, in combination with the retinoid X receptor (RXR) or retinoid A receptor (RAR), binds to its vitamin D response elements (VDRE) in the promoters of genes whose expression it regulates. The VDR also binds to one of two coactivator complexes, Mediator/DRIP (VDR interacting proteins) or p160/SRC (steroid hormone receptor complex), complexes which link the VDR to the RNA polymerase complex. We have recently discovered that the binding of VDR to these complexes is sequential. Binding to Mediator/DRIP occurs in the undifferentiated keratinocyte, but as the cell differentiates, DRIP(205) (the key protein of the DRIP complex binding to the VDR) levels fall, and p160/SRC binding takes over. We hypothesize that this sequential replacement of Mediator/DRIP by p160/SRC is critical for differentiation. Squamous cell carcinomas (SCC) fail to respond to the prodifferentiating actions of 1,25(OH)2D. These cells have normal levels of VDR and normal binding of VDR to VDREs. However, they fail to down-regulate DRIP(205) such that the p160/SRC complex fails to bind to VDR. This lack of sequential binding of these coactivator complexes to the VDR, we believe, maintains the cell in a state of continued proliferation and blocks the ability of 1,25(OH)2D to induce the expression of genes required for the differentiation process. PMID- 12899518 TI - Extrarenal sites of calcitriol synthesis: the particular role of the skin. AB - Calcitriol (1alpha,25(OH)2D3), the hormonally active form of vitamin D3 (D3) is produced by a cascade of reactions, including photochemical D3 synthesis in the skin and subsequent hydroxylation at the C-25 atom in the liver and finally at C 1alpha position in the kidney. However, there is substantial evidence for additional extrarenal sites of calcitriol synthesis. In vitro, many nonrenal cells, including bone, placenta, prostata, keratinocytes, macrophages, T lymphocytes and several cancer cells (e.g., from lung, prostata and skin) can enzymatically convert calcidiol (25OHD3) to 1alpha,25(OH)2D3. We have demonstrated that keratinocytes of the skin have unique properties in the D3 pathway; they are not only capable of producing D3 from 7-dehydrocholesterol (7 DHC), but also generate 1alpha,25(OH)2D3 from the substrates 25OHD3, lalpha hydroxyvitamin D3 (1alpha-OHD3) and even D3. It is evident that keratinocytes possess at least 1alpha-OH-, 25OH- and 24OHase activity, which is necessary for enzymatic conversion of D3 to 1alpha,25(OH)2D3, and the 24OHase activity for initiation of catabolism of 1alpha,25(OH)2D3 to more polar metabolites. Thus, the skin is apparently the only extrarenal tissue where the complete UVB-induced pathway from 7-DHC to 1alpha,25(OH)2D3 takes place under physiological circumstances. PMID- 12899517 TI - A novel vitamin D-regulated immediate-early gene, IEX-1, alters cellular growth and apoptosis. AB - 1alpha,25-Dihydroxyvitamin D3 (1alpha,25(OH)2D3) inhibits the expression of an immediate-early gene, IEX-1, which is involved in the regulation of cellular growth and apoptosis in a variety of cells. 1alpha,25(OH)2D3 alters the subcellular localization of IEX-1 by causing an efflux of IEX-1 from the nucleus, and the sterol decreases the expression of IEX-1 messenger RNA in cells via a novel DR3 repeat-type DNA response element. PMID- 12899519 TI - Cultured rat growth plate chondrocytes express low levels of 1alpha-hydroxylase. AB - Growth plate chondrocytes are the target of the hydroxylated vitamin D metabolites 1alpha,25(OH)2D3 and 24,25(OH)2D3. Because studies on the production of these polar metabolites were inconclusive in various in vitro systems, the expression of a potential paracrine/autocrine vitamin D system was examined in primary cultures of rat growth plate chondrocytes using real-time RT-PCR. Compared to UMR cells and renal homogenates primary cultures of growth plate chondrocytes expressed low levels of 25-hydroxy-1alpha-hydroxylase as well as 25 hydroxy-24-hydroxylase. The expression of both is modulated by 25 vitamin D3, but 1alpha,25(OH)2D3 affected only 25-hydroxy-24-hydroxylase. If these findings are confirmed in intact growth plates, the polar vitamin D metabolites could act in a paracrine/autocrine fashion within the growth plate. PMID- 12899520 TI - Gene amplification and splice variants of 25-hydroxyvitamin D3 1,alpha hydroxylase (CYP27B1) in glioblastoma multiforme--a possible role in tumor progression? AB - We report a gene amplification of 25-hydroxyvitamin D3 1,alpha-hydroxylase and the appearance of various splice variants in glioblastoma multiforme. Some protein products of the widespread expressed variants may exert a biological function. The analysis of amplification and overexpression of alternative transcripts will provide new insights in the regulation of the 25-hydroxyvitamin D3 1,alpha-hydroxylase expression and may have implications for prognosis in glioblastoma. PMID- 12899521 TI - Regulation of the 25-hydroxyvitamin D-1alpha-hydroxylase gene and its splice variant. AB - The 25-hydroxyvitamin D-1alpha-OHase (1alpha-OHase) is responsible for producing the active form of vitamin D, 1alpha,25-dihydroxyvitamin D. The enzyme not only is expressed in kidneys, but also is expressed in many nonrenal tissues, including skin. In this study, we compared the regulation of the 1alpha-OHase expression in kidney cells and keratinocytes. Using transfected luciferase reporter gene constructs, we compared the activity and regulatory features of the human 1alpha-OHase gene promoter in C-21 human kidney cells (PTH/PTHrP receptor positive) and cultured human keratinocytes (NHKs). We found that two regions, 1,100 bp and -396 bp from the ATG, were highly sensitive to parathyroid hormone (PTH) in C-21 cells but not in NHK. Furthermore, three CRE-like sequences (CLS) were identified within this PTH-sensitive area of the 1alpha-OHase promoter and when deleted they reduced induction of PTH by 50%-95% in C-21 cells. To further investigate the differential regulation profile, we examined the protein products of 1alpha-OHase in kidney and skin. Western blot analysis of whole cell extracts from these tissues with a 1alpha-OHase-specific antibody revealed the predicted 1alpha-OHase protein product of 56 kDa in kidney and a larger protein product of 59 kDa in skin. Using RT-PCR for the 1alpha-OHase in skin and kidney, we detected an insertion between exons 2 and 3 in skin but not in kidney. These results suggest that the regulation of renal and skin 1alpha-OHase gene expression may be tissue specific and possibly produce different splice variants, and that this specificity is likely conferred by differential expression of CRE-binding proteins in different cell types. In conclusion, the differential tissue expression of 1alpha-OHase gene variants and the tissue-specific regulation profile open up a new paradigm in the understanding of the role of 25 hydroxyvitamin D3 1alpha-hydroxylase gene in the regulation of vitamin D physiology. PMID- 12899522 TI - Combination of vitamin D metabolites with selective inhibitors of vitamin D metabolism. AB - 1alpha,25(OH)2D3 exerts antiproliferative, differentiating effects on many cell types, including cancer tissues. In most of its target cells, levels of 1alpha,25(OH)2D3 are regulated by local synthesis via CYP27B and metabolism via CYP24. Rapidly induced by vitamin D, CYP24 repeatedly hydroxylates the vitamin D side chain and ultimately terminates hormonal activity. Aiming at increased hormone levels, lifetime and function, numerous vitamin D analogs have been synthesized with structural modifications, which impede oxidation of the vitamin D side chain. Our group followed a different strategy, namely, blocking 1,25(OH)2D3 metabolism with inhibitors of CYP24. As appropriate inhibitors, we exploited compounds termed azoles, which directly bind to the heme iron of the CYPs via an azole nitrogen and to other parts of the substrate site. We synthesized some 400 azoles and tested their potential to selectively inhibit CYP24, but not hormone synthesis by the related CYP27B. Using primary human keratinocyte cultures as the source of CYP24 and CYP27, we discovered some 50 inhibitors of CYP24 with IC50 values in the nanomole range and selectivities up to 60-fold. As the first representative of selective CYP24 inhibitors, VID400 underwent preclinical development. In human keratinocytes, VID400 stabilized levels of endogenously produced 1alpha,25(OH)2D3, and thereby strongly amplified and prolonged expression of CYP24, a surrogate marker of hormonal function. In parallel, antiproliferative activity showed up at 100-fold or more lower concentrations of 1alpha,25(OH)2D3. This data suggests that CYP24 inhibitors could become attractive drugs in antiproliferative therapy, used as single entities to increase or extend endogenous hormone function or in combination with low doses of potent analogs. Moreover, we used selective inhibitors as valuable tools to (a) elucidate regulatory mechanisms of vitamin D synthesis and metabolism, (b) determine intrinsic activities of the otherwise highly transient vitamin D metabolites and (c) model the active sites of CYP24 and CYP27B. PMID- 12899523 TI - Importance of cytochrome P450-mediated metabolism in the mechanism of action of vitamin D analogs. AB - The elucidation of the metabolic pathway for vitamin D, including the delineation of the specific cytochrome P450s (CYPs) involved in activation and catabolism, has emphasized the overall importance of metabolic considerations in vitamin D analog design. This short review attempts to summarize recent findings with isolated CYPs and animal models in which CYPs are genetically manipulated to draw attention to structural features of vitamin D analogs that make them more or less resistant to metabolic enzymes. We conclude by placing metabolic considerations in the context of the other important aspects of vitamin D analogs. PMID- 12899524 TI - The role of vitamin D in prostate cancer. AB - Prostate cancer (PCa) cells harbor receptors for vitamin D (VDR) as well as androgens (AR). 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] increases AR expression and enhances androgen actions linking the two receptor systems. 1,25(OH)2D3 exhibits antiproliferative activity in both AR-positive and AR-negative PCa cells. Less calcemic analogs of 1,25(OH)2D3, with more antiproliferative activity, are being developed and will be more useful clinically. The mechanisms underlying differential analog activity are being investigated. In target cells, 1,25(OH)2D3 induces 24-hydroxylase, the enzyme that catalyzes its self inactivation. Co-treatment with 24-hydroxylase inhibitors enhances the antiproliferative activity of calcitriol. Primary cultures of normal or cancer derived prostatic epithelial cells express 1alpha-hydroxylase, the enzyme that catalyzes the synthesis of 1,25(OH)2D3, the levels being much lower in the cancer derived cells and in PCa cell lines. This finding raises the possibility of using 25-hydroxyvitamin D3 [25(OH)D3] as a chemopreventive agent in PCa. In LNCaP human PCa cells, 1,25(OH)2D3 and its analogs exert antiproliferative activity predominantly by cell cycle arrest, but also induce apoptosis, although to a much lesser degree. Growth arrest is mediated by induction of IGF binding protein-3 (IGFBP-3), which in turn increases the expression of the cell cycle inhibitor p21, leading to growth arrest. Other actions of 1,25(OH)2D3 in PCa cells include promotion of pro-differentiation effects and inhibition of tumor cell invasion, metastasis and angiogenesis. Combination therapy with retinoids, other anticancer agents or 24-hydroxylase inhibitors augments the inhibitory activity of 1,25(OH)2D3 in PCa and provides another effective approach in PCa treatment. Small clinical trials have shown that 1,25(OH)2D3 can slow the rate of prostate specific antigen (PSA) rise in PCa patients, demonstrating proof of concept that 1,25(OH)2D3 or its analogs will be clinically effective in PCa therapy. Current research involves further investigation of the role of 1,25(OH)2D3 and its analogs for the therapy or chemoprevention of PCa. PMID- 12899525 TI - Vitamin D autocrine system and prostate cancer. AB - 25-Hydroxyvitamin D-1alpha-hydroxylase (lalpha-OHase) is expressed in prostate cells. The expression suggests that local production of 1,25-dihydroxyvitamin D could provide an important cell growth regulatory mechanism. However, there is differential expression of 1alpha-OHase activity among the primary cultures of prostate cells derived from cancerous, benign prostatic hypertrophy and normal tissue, and among noncancerous (PZHPV-7) and various cancer cell lines (PC-3, DU145). No activity was found in cancer cell line LNCaP. The observed marked decrease in 1alpha-OHase activity in prostate cancer cells suggests some defect of the 1alpha-OHase in these cells. Using luciferase reporter gene assay, we observed a step-wise decrease in the basal promoter activity in two truncated promoter fragments, AN2 (-1,100 bp) and AN5 (-394 bp), with the highest basal activities found in PZHPV-7 and with loss of promoter activity in LNCaP. In order to understand the mechanism underlying the differential promoter activities among different prostate cells, we investigated the possible role of phosphorylation of cyclic AMP response element binding protein (CREB) on the regulation of 1alpha OHase promoter activity in the four prostate cell lines. First we compared the levels of CREB phosphorylation among PZHPV-7, DU145, PC-3 and LNCaP cells by Western blot analysis using antibody against phosphorylated CREB. We observed that CREB was phosphorylated to a greater extent in PZHPV-7 than in DU145 cells. No significant phosphorylation of CREB was found in PC-3 and LNCaP cells. Next, we utilized activators and inhibitors of protein kinase A (PKA), protein kinase C (PKC), mitogen-activated protein kinase kinase (MAPKK) and calcium/calmodulin dependent protein kinase II (CaMKII) to determine which kinases might be involved in phosphorylating the CREB in PZHPV-7 cells. We demonstrated that forskolin (an activator of PKA) increased the AN2 basal promoter activity 50%, whereas H-89 (an inhibitor of PKA) inhibited the basal and forskolin-stimulated AN2 promoter activity 40% and 70%, respectively. We also showed that PD98059 (an inhibitor of MAPKK) decreased the AN2 promoter activity 70%. Phorbol 12-myristate 13-acetate (an activator of PKC), GF109203 (an inhibitor of PKC) and KN-93 (an inhibitor of CaMKII) had no effect on AN2 promoter activity in PZHPV-7 cells. Thus, our results suggest that differential phosphorylation of CREB through PKA and MAPK pathways may be involved in the regulation of 1alpha-OHase promoter activity. PMID- 12899526 TI - Analysis of the vitamin D system in cervical carcinomas, breast cancer and ovarian cancer. AB - 1,25-Dihydroxyvitamin D3 (1,25(OH)2D3) is the biologically active metabolite of vitamin D and has been shown to regulate the growth of various cell types. There are two principal enzymes involved in the formation of circulating 1,25(OH)2D3 from vitamin D, the vitamin D 25-hydroxylase (25-OHase) and the 1alpha hydroxylase (1alpha-OHase). Recently, extrarenal activity of 1alpha-OHase has been reported in various cell types. The aim of this study was to analyze expression of VDR and the main enzymes involved in the synthesis and metabolism of calcitriol in gynecological malignancies and corresponding normal tissue. Expression of VDR, 25-OHase, 1alpha-OHase, and 24-OHase was analyzed in breast carcinomas (BC), ovarian cancer (OC), cervix carcinomas (CC) and normal corresponding tissues using real-time PCR and specific hybridization probes as well as using immunohistochemistry. RNA for VDR, 1alpha-OHase, 24-OHase and 25 OHase was up-regulated in breast cervical and ovarian carcinomas as compared to normal tissue. VDR immunoreactivity was increased in breast and ovarian cancer and in cervix carcinomas as compared to normal corresponding tissue. Our findings indicate that cervical carcinomas, breast cancer and ovarian cancer may be considered as potential targets for prevention or therapy with new vitamin D analogs that exert little or no calcemic side effects or by pharmacological modulation of 1,25(OH)2D3 synthesis and metabolism in these tumor cells. PMID- 12899527 TI - Short-chain fatty acids and colon cancer cells: the vitamin D receptor--butyrate connection. AB - Butyrate and its prodrug tributyrin, as well as 1,25-dihydroxyvitamin D3 (1,25 (OH)2D3), have important physiological effects on proliferation and differentiation in a variety of malignant cells. The aim of this study was to elucidate the role of the vitamin D receptor (VDR) in butyrate-induced cell differentiation and cell cycle arrest in Caco-2 cells, a human colon cancer cell line. Cell differentiation was evaluated by analyzing the activity of alkaline phosphatase (AP). Protein of VDR, cyclins, cyclin-dependent kinases (cdks) and of cdk inhibitors was quantified by Western blot analysis, VDR-mRNA by PCR. Pre- and postconfluent cells were assessed for VDR binding activity. Cell cycle was analyzed by flow cytometry. Tributyrin significantly increased VDR-mRNA level (250% vs. control) and VDR binding activity. Butyrate also enhanced VDR protein content in the nucleus in a time- and dose-dependent manner and more potently than other short-chain fatty acids of a related structure. Both butyrate (640% vs. control) and 1,25-(OH)2D3 (350% vs. control) significantly stimulated differentiation, whereas combined treatment with butyrate and 1,25-(OH)2D3 resulted in a synergistic amplification of AP activity (1400% vs. control). In the presence of the VDR antagonist ZK 191732, butyrate-induced differentiation was completely abolished (150% vs. control). While butyrate alone increased p21Waf1/Cip1 expression and down-regulated cdk 6 and cyclin A, and combined exposure with 1,25-(OH)2D3 resulted in a synergistic enhancement of butyrate induced changes, expressions did not change from control level after treatment with butyrate and ZK 191732. G1 cell cycle arrest induced by butyrate was also abolished after combined treatment with butyrate and ZK 191732. In conclusion, differentiation and cell cycle arrest of Caco-2 cells induced by butyrate are mediated by up-regulation of VDR, followed by a stimulation of the negative cell cycle regulator p21Waf1/Cip1 and by a down-regulation of cdk 6 and cyclin A, both involved in cell cycle progression. PMID- 12899528 TI - Analysis of the vitamin D system in cutaneous malignancies. AB - 1,25-dihydroxyvitamin D3, the biological active metabolite of vitamin D, has great impact on keratinocyte growth and differentiation, and consequently has already been successfully used in the therapy of hyperproliferative skin disorders. We have now characterized the key components of the vitamin D system (VDR, 1alpha-OHase, 24-OHase and 25-OHase) in cutaneous basal cell carcinomas (BCC) and squamous cell carcinomas (SCC), using immunohistochemical and quantitative real-time PCR techniques. Additionally, proliferative activity (Ki 67 expression), differentiation status (cytokeratin 10 and transglutaminase K expression), rate of apoptosis (TUNEL assay) and the abundance of the main heterodimerization partners of VDR (RXRs) was determined for these tumours and correlated with the components of the Vitamin D system. Our findings indicate that the Vitamin D system may be of high importance for the growth behaviour of BCCs and SCCs and that new vitamin D analogues that exert less calcaemic side effects may be effective in the prevention or treatment of these tumours. PMID- 12899529 TI - Evaluation of vitamin D analogs as therapeutic agents for prostate cancer. AB - Prostate cancer cells contain specific receptors (VDR) for la,25-dihydroxyvitamin D (1alpha,25(OH)2D), which is known to inhibit the proliferation and invasiveness of these cells. These findings support the use of 1alph,25(OH)2D for prostate cancer therapy. However, because 1alpha,25(OH)2D can cause hypercalcemia, analogs of 1alpha,25(OH)2D that are less calcemic but which exhibit potent antiproliferative activity would be attractive as therapeutic agents. We studied four vitamin D compounds: 25-hydroxyvitaminD3 [25(OH)D3], which is converted to 1alpha,25(OH)2D3 in prostate cells, and three analogs of 1alpha,25(OH)2D3: EB1089, 19-nor-1alpha,25(OH)2D2 and hexafluoro-1alpha,25(OH)2D3 (F6 1alpha,25(OH)2D3). 19-nor-1alpha,25(OH)2D2 has been shown to be less calcemic than 1alpha,25(OH)2D3 in clinical trials. F6-1alpha,25(OH)2D3 has been shown to be 100-fold more active than 1alpha,25(OH)2D3 and to be longer-lasting in inhibiting keratinocyte proliferation in vitro. EB1089 has been shown to be less calcemic than 1alpha,25(OH)2D3 in rats implanted with Leydig cell tumors. For 25(OH)D3, 19-nor-1alpha,25(OH)2D2 and F6-1alpha,25(OH)2D3, we studied the in vitro effects and compared their activity to 1alpha,25(OH)2D3 on cellular proliferation by 3H-thymidine incorporation assay. In addition, we studied transactivation of the VDR in the presence of 25(OH)D3 and 19-nor 1alpha,25(OH)2D2 in prostate cells. For EB1089, we compared its inhibition of prostate cancer metastasis to that induced by 1alpha,25(OH)2D3 in vivo in the rat Dunning MAT LyLu prostate cancer model. We found that 1alpha,25(OH)2D3 and 19-nor 1alpha,25(OH)2D2 caused similar dose-dependent inhibition in 3H-thymidine incorporation into DNA in prostate cells and behaved similarly in the CAT reporter gene transactivation assay in PC-3/VDR cells. F6-1alpha,25(OH)2D3 is 10- to 50-fold more active than 1alpha,25(OH)2D3 in 3H-thymidine incorporation into DNA in the primary cultured prostate cells. Likewise, 25(OH)D3 had comparable antiproliferative activity to la,25(OH)2D3. In the rat model, tumor volumes and the number of metastases in the lungs were significantly reduced by both 1alpha,25(OH)2D3 (10.4 +/- 2.81 tumor foci) and EB1089 (7.7+/-1.29 tumor foci) compared to controls (22.7 +/- 1.98 tumor foci). Although serum calcium levels were significantly elevated in both 1alph,25(OH)2D3- and EB1089-treated rats, EB1089 was significantly less calcemic than 1alpha,25(OH)2D3 (12.59+/-0.21 mg/dl versus 14.47+/-.46 mg/dL; 1 microg/kg; p < 0.001). In conclusion, our data indicate that 25(OH)D3 and the three 1alpha,25(OH)2D analogs represent two different solutions to the problem of hypercalcemia associated with vitamin D based prostate cancer therapies: 25(OH)D3 requires the presence of 25 hydroxyvitaminD-1alpha-hydroxylase, whereas 19-nor-1alpha,25(OH)2D2, F6 1alpha,25(OH)2D3 and EB1089 do not. These compounds may be good candidates for human clinical trials in prostate cancer. PMID- 12899530 TI - Design, synthesis, and biological studies of the A-ring-modified 1,25 dihydroxyvitamin D3 analogs. AB - Antitumor effects of 1alpha,25-dihydroxyvitamin D3 analogs have recently become one of the major topics of the vitamin D research field. We focused on the structure-activity relationships of the A-ring moiety of the vitamin D molecule and found several strong agonists of the vitamin D receptor, using a design of introducing a functional group into the C2 position. In the first step, all eight possible diastereomers of novel 2-methyl-1,25-dihydroxyvitamin D3 were synthesized using the convergent method with palladium catalyzed coupling reaction. We studied conformational analysis of each isomer based on 1H NMR and computer calculations; and biologically, VDR binding affinity, potency of induction of HL-60 cell differentiation, and apoptosis were investigated in detail. The biological effect of double modification in a combination of the CD ring side chain (20-epi, 20-epi-22R-methyl, and KH-1060 types) and the 2-methyl group was then evaluated. In this context, 5,6-trans derivatives of 2-methyl analogs were also synthesized and tested. Through these experiments, our accumulated knowledge that the 2a-methylated analog with the natural la,3fl dihydroxyl groups possesses a strong and unique biological profile guided us the next synthetic goal, i.e., three kinds of longer functional groups: 2alpha-alkyl, 2alpha-hydroxyalkyl, and 2alpha-hydroxyalkoxyl groups, which were introduced into 1alpha,25-dihydroxyvitamin D3, stereoselectively. We found that five of our new 2alpha-modified analogs show higher VDR-binding affinity than that of the natural hormone. HL-60 cell differentiation induction activities and calcium mobilization were studied for some of these compounds. These are the first examples, including the pioneer 2a-methyl analog, that exhibit higher VDR-binding affinity than 1alpha,25-dihydroxyvitamin D3 with pure A-ring modifications. To explain the effect, docking studies of the synthetic ligands to VDR are also described. This study could stimulate the development of antitumor medicines of the vitamin D analogs. PMID- 12899531 TI - Induction of apoptosis by vitamin D metabolites and analogs in a glioma cell line. AB - Gliomas are the most common malignant tumors in brain. Recent studies demonstrate the capacity of 1alpha,25(OH)2D3 to specifically induce cell death (apoptosis) in model glioma cell lines and in primary cultures from tumor tissue, but not in primary astrocytes. In spite of this promising activity, a broad therapeutic application of vitamin D metabolites and analogs is still restricted because of their poor bioavailability and their hypercalcemic actions. Compared to 1alpha,25(OH)2D3, its natural 3alpha-epimer exhibits far higher metabolic stability and a reduced calcemic effect. Focusing on a possible therapeutic advantage of the 3alpha-conformation, we have examined the apoptotic potential of a representative set of vitamin D analogs, each of them in the 3alpha- and 3beta conformation, and of natural vitamin D metabolites in the rat C6 glioma cell line. Exposure of these cells to the synthetic analogs resulted in all cases in a pronounced reduction of cell density (tested by incorporation of neutral red) and induction of apoptosis, monitored by staining nuclei with Hoechst 33258 dye and by following DNA fragmentation by capillary electrophoresis. The 3alpha-epimers showed equivalent or even higher activity on C6 cells than their respective 3beta forms. For their potent effects on growth and apoptosis of tumor cells and their high metabolic stability combined with a low calcemic potential, we speculate that these 3a-epimers could provide advantages for a prospective treatment of glioma. PMID- 12899532 TI - Vitamin D analogs and breast cancer. AB - Breast cancer is the most frequent malignancy of women in the Western world. Vitamin D compounds constitute a novel alternative to the conventional use of antiestrogens for chemoprevention and chemotherapy. The biologically active form of vitamin D, 1,25-Dihydroxyvitamin D3 [1,25(OH)2D3], not only plays an essential role in the control of calcium homeostasis, but also acts on cells of a variety of tissues to promote inhibition of cellular proliferation and induction of differentiation. The potential use of 1,25(OH)2D3 in the treatment of cancer is limited by its propensity to cause hypercalcemia at pharmacologically active doses. This has led to the synthesis of analogs of vitamin D that exhibit potent anticancer effects, but have low calcemic activity. Evidence from both in vitro and in vivo studies has demonstrated that vitamin D compounds can inhibit the growth of breast cancer cells, suggesting their therapeutic value in the treatment or prevention of this disease. PMID- 12899533 TI - A low-calcemic vitamin D analog (Ro 25-4020) inhibits the growth of LNCaP human prostate cancer cells with increased potency by producing an active 24-oxo metabolite (Ro 29-9970). AB - In this study, we have characterized a novel less-calcemic vitamin D analog Ro 25 4020 (1alpha, 25 dihydroxy-16-ene-5,6-trans-vitamin D3) and investigated the mechanisms underlying its enhanced growth inhibitory properties. We found that Ro 25-4020 (IC50 = 0.3 nM) exhibited greater inhibitory activity than 1,25(OH)2D3 (IC50 = 1 nM) on LNCaP human prostate cancer cell growth. However, Ro 25-4020 was tenfold less active than 1,25(OH)2D3 in receptor-binding assays, ligand-induced heterodimerization and transactivation assays using VDR. HPLC and GC-MS analyses revealed that 1,25(OH)2D3 is converted to a 24-hydroxy metabolite, which has been shown to be less potent than 1,25(OH)2D3. In contrast, Ro 25-4020 was converted to a major 24-oxo metabolite that was more stable. Ligand-binding assays reveal that both Ro 25-4020 and its 24-oxo metabolite have similar affinity for VDR. Synthetic 24-oxo-Ro 25-4020, however, inhibited LNCaP cell proliferation as potently as 1,25(OH)2D3 and was more potent in transactivation of two out of three vitamin D target genes tested. These results suggest that conversion of Ro 25-4020 into an active and more stable 24-oxo metabolite with longer half-life contributes significantly to its potent antiproliferative actions on the LNCaP cells. PMID- 12899534 TI - Prospects for vitamin D receptor modulators as candidate drugs for cancer and (auto)immune diseases. AB - The development of less calcemic vitamin D analogs creates possible therapeutic applications for immune modulation (e.g., autoimmune diseases and graft rejection), inhibition of cell proliferation and induction of cell differentiation (e.g., cancer). Recently more insight was obtained in the mechanism of action of the analogs at the biological and molecular level. Critical remarks are summarized on why the step towards the clinic has not yet been taken and how better selective vitamin D receptor modulators could be designed. PMID- 12899535 TI - The role of reactive oxygen species in the anticancer activity of vitamin D. AB - Calcitriol, the hormonal form of vitamin D, enhances the anticancer activity of the immune cytokine tumor necrosis factor, interleukin 1 and interleukin 6 in human breast and renal cell carcinoma cells without affecting the cytotoxic action of interferon-alpha or killer lymphocytes. It also enhances cytotoxicity induced by the anticancer drug doxorubicin, by the redox cycling quinone menadione, and by the reactive oxygen species hydrogen peroxide. The synergistic interaction was accompanied by increased oxidative stress, as manifested by glutathione depletion and was abolished by exposure to the thiol antioxidant N acetylcysteine. The hormone on its own brought about an increase in the cellular redox state as reflected in the ratio between oxidized and reduced glutathione and glyceraldehyde-3-phosphate dehydrogenase, and a reduction in the expression of the antioxidant enzyme Cu/Zn superoxide dismutase. These results support the notion that the interplay between active vitamin D derivatives and other anticancer agents such as immune cytokines and anticancer drugs plays a role in the in vivo anticancer activity of vitamin D and that reactive oxygen species are involved in the anticancer activity of vitamin D on its own and in its cross-talk with other anticancer modalities. PMID- 12899536 TI - Ecologic studies of solar UV-B radiation and cancer mortality rates. AB - Solar ultraviolet B (UV-B) radiation (280-320 nm) has been associated with reduced risk of cancer of the breast, colon, ovary, and prostate, as well as non Hodgkin's lymphoma (NHL) through the production of vitamin D in papers extending back to 1980. Using data on the geographic distribution of cancer mortality rates in the US, another ten cancers have been added to the list for which UV-B/vitamin D is a risk reduction factor (Grant 2002b; submitted). These associations persist even after additional cancer risk and risk reduction factors such as smoking, urban or rural residence, Hispanic heritage, poverty, dietary factors, and use of nonsteroidal anti-inflammatory drugs are added to the analysis. As a further test of the protective role of UV-B radiation, an ecologic study of cancer mortality rates in Europe with UV-B radiation and dietary factors was conducted. Inverse correlations are found for UV-B radiation for a number of cancers, with those for bladder, breast, endometrial, ovarian, prostate, and renal cancer, and multiple myeloma and NHL having the strongest correlations in this and ongoing multicountry ecologic studies. These studies add further support for the role of UV-B radiation and vitamin D in reducing the risk of a large number of cancers. PMID- 12899537 TI - Phytoestrogens and 17beta-estradiol influence vitamin D metabolism and receptor expression-relevance for colon cancer prevention. AB - 1alpha,25(OH)2D3 is a potent growth inhibitor of different cancer cell lines. The steroid hormone is not only synthesized in the kidney, but also at extrarenal sites. Unfortunately, this potential autocrine/paracrine defense mechanism is lost during the late stages of colon tumor progression. It is therefore desirable to find a pharmacological means to maintain or enhance endogenous production of 1alpha,25(OH)2D3 during early periods in tumorigenesis. The phytoestrogen genistein was shown to regulate different cytochrome P450 enzymes, a family of proteins to which both of the vitamin D-metabolizing CYP27B1 (1alpha-hydroxylase) and CYP24 (24-hydroxylase) belong. Therefore, we used two colon cancer cell lines, Caco-2 and COGA-1, and investigated possible influences of genistein on different parameters of extrarenal vitamin D metabolism by HPLC, RT-PCR, and Western blot analysis. Differences between the two cell lines were found in both their basic enzymatic activities and in their response to treatment with 1alpha,25(OH)2D3. Whereas Caco-2 cells responded to administration of 100 nM genistein with a down-regulation of 24-hydroxylase activity, COGA-1 cells showed not only a significant down-regulation of 24-hydroxylase protein expression, but also a clear induction of vitamin D receptor (VDR) expression. Similar effects on VDR expression were achieved by administration of 10 nM 17beta-estradiol. This suggests an estrogenic mode of action of genistein, which might be dependent on differential distribution of estrogen receptors alpha and beta in our cell lines. PMID- 12899538 TI - Efficacy and mechanism of action of 1alpha-hydroxy-24-ethyl-cholecalciferol (1alpha[OH]D5) in breast cancer prevention and therapy. AB - It is now well established that the active metabolite of vitamin D3, 1alpha,25(OH)2D3, regulates cell growth and differentiation in various in vitro cancer models. However, its clinical use is precluded due to its hypercalcemic activity in vivo. Hence, several less calcemic vitamin D analogs have been synthesized and evaluated for their chemopreventive and therapeutic efficacy in experimental carcinogenesis models. A novel analog of vitamin D3, 1alpha-hydroxy 24-ethyl-cholecalciferol (1alpha[OH]D5), has currently been under investigation in our laboratory for its application in breast cancer prevention and therapy. 1alpha(OH)D5 had been shown to inhibit development of estrogen- and progesterone dependent ductal lesions as well as steroid hormone-independent alveolar lesions in a mammary gland organ culture (MMOC) model. Moreover, the inhibitory effect was more significant if 1alpha(OH)D5 was present during the promotional phase of the lesion development. The growth inhibitory effect of 1alpha(OH)D5 has also been manifested in several breast cancer cell lines, including BT-474 and MCF-7. Breast cancer cell lines that responded to 1alpha(OH)D5 treatment were vitamin D receptor positive (VDR+). Vitamin D receptor-negative (VDR-) cell lines, such as MDA-MB-231 and MDA-MB-435, did not show growth inhibition upon incubation with 1alpha(OH)D5. This suggests the requirement of VDR in 1alpha(OH)D5-mediated growth effects. Interestingly, breast cancer cells that were VDR+ as well as estrogen receptor positive (ER+) showed cell cycle arrest and apoptosis, while VDR+ but ER- cells (UISO-BCA-4 breast cancer cells) showed enhanced expression of various differentiation markers with la(OH)D5 treatment. Transcription and expression of estrogen-inducible genes, progesterone receptor (PR) and trefoil factor 1 (pS2), were significantly down-regulated in ER+ BT-474 cells with 1alpha(OH)D5 treatment. This implies a differential effect of 1alpha(OH)D5 on ER+ vs. ER- cells. Additionally, comparison between the effects of 1alpha(OH)D5 on normal vs. transformed cells indicated that 1alpha(OH)D5 does not suppress cell prolifera- PMID- 12899539 TI - Regulation of extrarenal vitamin D metabolism as a tool for colon and prostate cancer prevention. AB - Though 1,25-dihydroxyvitamin D3 (1,25-D3) as well as some vitamin D analogs have an antimitotic as well as a differentiating action, therapeutic application in tumor patients is still precluded due to their hypercalcemic action at the necessary concentration. Our observation that early during progression, colon tumor cells express CYP27B1, the enzyme essential for 1,25-D3 synthesis, as well as the vitamin D receptor (VDR) at a higher level than normal colon cells led to the speculation that, by induction of this expression, a physiological defense against tumor progression could be activated and enhanced. In some Asian countries where soy products are a main staple, prostate and breast tumor incidence is extremely low. We speculated that this could be due to regulation of CYP enzymes by phytoestrogens present in soy such as genistein. In prostate tumor cells, the 1,25-dihydroxyvitamin D3 catabolizing enzyme CYP24 is frequently highly expressed. We were able to demonstrate that genistein down-regulates expression of CYP24 to almost nil, which would result in enhancement of local 1,25-D3 levels and improved mitotic control of tumor cells. PMID- 12899540 TI - Modulation of X-ray-induced apoptosis in human keratinocytes (HaCaT) by 1,25 dihydroxyvitamin D3. AB - Possible effects of 1,25-dihydoxyvitamin D3 (vitamin D) on ionizing radiation induced cell damage have been unknown until now. The task of the present study was to analyze, in a human keratinocyte cell line (HaCaT), the effects of a preincubation with vitamin D on the X-ray-induced mRNA expression of different genes related to apoptosis (gene array). The first results show that ionizing radiation leads to a down-regulation of various apoptosis-relevant genes in HaCaT cells pretreated with vitamin D. Therefore it can be speculated that vitamin D could prove to be a promising radioprotective substance. PMID- 12899541 TI - The effects of hydroxyapatite coating and bone allograft on fixation of loaded experimental primary and revision implants. AB - We used our established experimental model of revision joint replacement to examine the roles of hydroxyapatite coating and bone graft in improving the fixation of revision implants. The revision protocol uses the Soballe micromotion device in a preliminary 8-week period of implant instability for the presence of particulate polyethylene. During this procedure, a sclerotic endosteal bone rim forms, and a dense fibrous membrane is engendered, having macrophages with ingested polyethylene and high levels of inflammatory cytokines. At the time of revision after 8 weeks, the cavity is revised with either a titanium alloy (Ti) or a hydroxyapatite (HA) 6.0 mm plasma-sprayed implant, in the presence or absence of allograft packed into the initial 0.75 mm peri-implant gap. The contralateral limb is subjected to primary surgery with the same implant configuration, and serves as control. 8 implants were included in each of the 8 treatment groups (total 64 implants in 32 dogs). The observation period was 4 weeks after revision. Outcome measures are based on histomorphometry and mechanical pushout properties. The revision setting was always inferior to its primary counterpart. Bone graft improved the revision fixation in all treatment groups, as also did the HA coating. The sole exception was revision-grafted HA implants, which reached the same fixation as primary Ti and HA grafted implants. The revision, which was less active in general, seems to need the dual stimulation of bone graft and HA implant surface, to obtain the same level of fixation associated with primary implants. Our findings suggest that the combination of HA implant and bone graft may be of benefit in the clinical revision implant setting. PMID- 12899542 TI - Good performance of a titanium femoral component in cementless hip arthroplasty in younger patients: 97 arthroplasties followed for 5-11 years. PMID- 12899543 TI - Cementless Spotorno tapered titanium stems: excellent 10-15-year survival in 141 young patients. AB - We evaluated the clinical and radiographic results of the first consecutive 154 implantations of a cementless, double-tapered straight femoral stem (cementless Spotorno (CLS), Sulzer Orthopedics) in 141 patients under the age of 55 (mean 47 (13-55)) years. After a mean follow-up of 12 (10-15) years, 11 patients (11 hips) had died and 7 (7 hips) could not be located. 5 patients (5 hips) underwent femoral revision-1 for infection, 1 for periprosthetic fracture and 3 for aseptic loosening of the stem. The overall survival rate of the stem was 97% at 12 years (95% confidence limits, 93%-100%), and survival with femoral revision for aseptic loosening as an end point 98 (95-100)%. The survival rate of the acetabular components was 78 (71-85)% after 12 years. The median Harris hip score at follow up was 84 points. None of the patients had thigh pain. Radiolucent lines in Gruen regions 1 and 7 were present in 21 hips (17%). 2 hips had radiolucent lines in regions 2-6 on anteroposterior (AP) radiographs. No femoral osteolysis was detected. The mid- to long-term survival with this type of femoral component is excellent and compares favorably with cemented stems in this age group. However, the high rate of cup loosening and the low Harris hip scores are a concern in this subgroup of young patients. PMID- 12899544 TI - Comparison of hydroxyapatite- and porous-coated stems in total hip replacement. AB - 24 patients underwent bilateral hip replacement, using the SROM modular system (Depuy Johnson & Johnson, MA, USA) with the porous-coated sleeve in one hip and an HA-coated sleeve in the other. 20 patients were followed for at least 4 years with clinical and radiographic evaluations. We found no difference between the two sides as regards the time of disappearance of thigh pain and hip scores. Radiographs showed a buttress sign in 18 HA-coated and 15 porous coated stems. There were no differences in the serial bone-remodeling pattern around the proximal sleeve and measurement of wear volume between the two groups. In the short- to medium-term, our findings support the view that the hydroxyapatite coated femoral stem has no advantages over those with porous coating. PMID- 12899545 TI - Effectiveness of the ROBODOC system in preventing intraoperative pulmonary embolism. AB - Intraoperative pulmonary embolism occurs not only during cemented but also during cementless total hip arthroplasty (THA). We determined whether the ROBODOC femoral milling system can reduce intraoperative pulmonary embolism, by using of transesophageal echocardiography and hemodynamic monitoring. We did a prospective clinical trial with 71 patients (75 hips) who were divided into 2 groups: group 1, 46 patients (50 hips) who underwent cementless THA with preparation of the femoral canal using ROBODOC; group 2, 25 patients (25 hips) who underwent conventional cementless THA surgery in whom separate measurements were made during preparation of the femur, insertion of the stem and relocation of the hip. The incidence of severe embolic events was lower in group 1 than in group 2. Our findings suggest that the ROBODOC femoral milling system may reduce the risk of clinically significant pulmonary embolism during cementless THA. PMID- 12899546 TI - Robotic bone preparation does not increase cement penetration into the proximal femur: a matched-pair cadaver study comparing hand-broaching versus robotic bone preparation. AB - In a cadaver study, we prepared 20 matched pairs of human femora using chipped tooth broaches and robotic milling with the same geometry. For robotic bone preparation the CASPAR robotic system with a rotating milling head was used. Cancellous bone was irrigated with 1 liter of pulsed lavage and the specimens were embedded in specially-designed pots. After vacuum mixing, bone cement was introduced in a retrograde manner and subjected to a standard pressure protocol with a constant force of 3,000 N. Radiographs were taken and horizontal sections were obtained at predefined levels, using a diamond saw. Microradiographs of the bone slices were taken, digitized and analyzed to assess cement penetration into cancellous bone. No femoral fractures or fissures occurred with either preparation technique. The microradiographic evaluation showed no morphometric differences between chipped-tooth broaches and robotic milling as regards cement penetration into cancellous bone. Therefore, in the presence of pulsed lavage, we conclude that robotic bone preparation does not increase cement penetration into cancellous bone of the proximal end of the femur. PMID- 12899547 TI - Salter's innominate osteotomy for hip dysplasia in adolescents and young adults: results in 58 patients (69 osteotomies) at 4-12 years. AB - We reviewed 69 Salter's innominate osteotomies (SIO) in 58 patients with developmental dysplasia of the hip. Their mean age at operation was 22 (13-37) years. 24 hips were operated on in patients under 18 years of age (adolescents) and 45 hips in patients 18 years or over (adults). The mean follow-up was 7.6 (3.8-12) years. The mean Harris hip score (HHS)/Merle d'Aubigne score improved from 66 (38-97)/14.0 (8.0-18) points before surgery to 81 (22-100) points/15 (6 18) points at final review. Using the endpoints total hip arthroplasty and a Merle d'Aubigne score of < 13 and/or a HHS < 71, 13 of the 69 hips had failed and the cumulative survival rate was 0.79 (95% CI, 0.69-0.89) at 12 years. Their age at operation and the grade of preoperative coxarthrosis affected the clinical outcome. No deterioration in the mean grade of coxarthrosis was noted at final review, as compared to the condition before surgery. We conclude that the SIO is a well-standardized operation, with a low risk of complications, which gives encouraging results in adolescents and young adults with or without mild arthrosis. The operation may retard or even arrest the coxarthrosis. PMID- 12899548 TI - Coverage of the femoral head by the acetabular labrum in dysplastic hips: quantitative analysis with radial MR imaging. AB - Hip dysplasia is an important cause of osteoarthrosis of the hip, especially in Japanese. Recently, the role of the acetabular labrum in hip diseases has attracted the attention of researchers, but the condition of the labrum in the dysplastic hips has not been described. We used radial MRI to measure the degree of coverage of the femoral head by the acetabulum and the labrum, corresponding to 7 positions on the acetabular rim at every 15 degrees from anterosuperior 45 degrees via midsuperior to posterosuperior 45 degrees. 38 Japanese patients (51 hips) with dysplasia, and 11 healthy controls 122 hips) were studied. In the dysplastic hips, the labrum was larger than in the controls. We found no correlation between the coverage of the acetabulum in the anterosuperior positions and the size of the labrum among the dysplastic hips. PMID- 12899549 TI - Costs of internal fixation and arthroplasty for displaced femoral neck fractures: a randomized study of 68 patients. AB - We included in a prospective, randomized study 68 patients aged 70 years or older, with displaced cervical hip fractures. The patients were randomized to internal fixation with hook-pins (36) or primary arthroplasty (32) (total or hemiarthroplasty due to their prefracture status) and followed for 2 years. Patients with rheumatoid arthritis, mental confusion and/or residence in an institution were excluded. The postoperative stay in hospital, rehabilitation wards or nursing homes were recorded as well as complications and the costs of surgery. The aim of this study was to compare the accumulated costs of each method, during the first 2 years after the fracture. In the internal fixation group, 15/36 were considered failures, as compared to 1/32 in the arthroplasty group. As regards primary treatment of the fracture, the durations of surgery and hospital stay were shorter after internal fixation, but the total need for hospitalization/institutionalization was somewhat longer in these patients. The mean 2-year cost for a patient with internal fixation was USD 21,000 and of one with primary arthroplasty USD 15,000. We conclude that primary arthroplasty is a cost-efficient treatment. Considering the very much higher failure rate after internal fixation--leading to increased suffering for these patients--primary arthroplasty stands out as the best method for displaced fractures of the femoral neck. PMID- 12899550 TI - Risk of clinical pulmonary embolism after joint surgery in patients receiving low molecular-weight heparin prophylaxis in hospital: a 10-year prospective register of 3,954 patients. AB - We studied the incidence of nonfatal, radiologically-confirmed, clinical pulmonary embolism (PE) after major joint surgery during 10 years of observation. The findings are based on a prospective register of all patients undergoing total hip replacement (THR), total knee replacement (TKR), or nailed hip fracture (NHF) in a Scandinavian hospital between 1989 and 1998. All patients received thromboprophylaxis with low-molecular-weight heparin, continued until discharge. Patients with suspected PE underwent ventilation/perfusion scintigraphy and/or spiral CT. Patients with concomitant clinical signs of deep vein thrombosis (DVT) were also subjected to imaging diagnostics. 3,954 patients underwent THR, TKR, or NHF; 122 of them were readmitted on clinical suspicion of PE, and 50 cases were confirmed. Of patients with confirmed PE, 6/50 had DVT. The average time to readmission was 35 (5-94) days after THR, 24 (1-173) days after NHF, and 9 (2-17) days after TKR. Following major hip surgery, the incidence of PE remained high for at least 2-3 months (less following TKR) in those given thromboprophylaxis for about 10 days. The differences in PE incidence and the time when it developed in NHF versus THR and TKR patients suggest that these patients should be considered separately when determining the optimal thromboprophylactic regimen. PMID- 12899551 TI - Muscle strength in children treated for displaced femoral fractures by external fixation: 31 patients compared with 31 matched controls. AB - In a prospective study (1993-2000), we measured the isokinetic strength of the quadriceps and hamstring muscles in 31 children aged 5-17 years, on average, 3 (1.5-5) years after treatment for a displaced femoral fracture by external fixation and early mobilization. A group of age-, sex- and weight-matched children without previous injury were used as controls. The hop-index test was used to assess the patient's confidence in the injured limb and was similar in the fractured and unfractured legs as well as in the patients and controls. We measured the peak torque output at 2 angular velocities (60 degrees/s and 180 degrees/s) in the hamstring and quadriceps muscles, using Cybex testing equipment. Torque to body weight ratios were used to compare muscle strength in patients and controls. We found no differences in muscle strength between patients and controls or in the distribution of which leg was stronger, equal or weaker in the patients or controls at any test speed. External fixation and early mobilization seem to prevent residual muscle weakness, which occurs with traction or a cast for femoral fractures in children. PMID- 12899552 TI - Transverse process wiring for thoracic scoliosis: a new technique. AB - We describe a method of reducing neurological risk by avoiding neural elements when performing segmental spinal fixation for scoliosis. This was done by making use of the transverse processes of the thoracic vertebrae 1-11 and wired plates. The wires are prevented from cutting the bone by attaching them to rectangular plates. These wired plates on the two sides of each segment should be firmly fixed to the base of the transverse process by tying them to each other. Maximum correction is obtained by spreading the force applied by stabilizing each thoracic vertebra from both sides. Derotation is attempted when all the wires have been tightened. Tension forces are spread to all fixated vertebrae simultaneously and the correction should be carried out at all levels at the same time. On the convex side, each wire connects over the rod to the next one above it and, tightened one by one, provides compression. We treated 15 patients having a thoracic curve with this method and followed them for 25-39 months. In 2 cases of congenital scoliosis, a 50% correction was obtained. In 3 cases of neurofibromatosis scoliosis, a 55% correction was achieved. In 10 cases of idiopathic scoliosis, the correction was 60%. When the thoracic sagittal contour was evaluated, thoracic kyphosis, which had been between -20 degrees and +90 degrees, had improved to between +8 degrees and +43 degrees in all cases. No infection or neurological complications occurred. PMID- 12899553 TI - Attachment of autogenous tendon graft to cortical bone is better than to cancellous bone: a mechanical and histological study of MCL reconstruction in rabbits. AB - We analyzed the mechanical and histological variables after the attachment of an autogenous tendon graft to cortical or cancellous bone. We reconstructed the medial collateral ligament of the knee in 33 Japanese white rabbits, using a bone socket procedure. The floor of the bone socket was cortical bone in group A and cancellous bone in group B. Mechanically, the pull-out test showed a tendency towards an increase in maximum failure load, with 10.9 N, 35 N and 37 N in group A, and 11 N, 18 N and 36 N in group B at 2, 4 (statistically significant difference) and 8 weeks after surgery, respectively. Histologically, the attachments were immature at 2 weeks. At 4 weeks, granulations had matured and Sharpey's fiber-like structures were seen. These fibers were more abundant in group A than in group B. At 8 weeks, the attachments in both groups were rather like the normal 4-zone structure. With time, tendon attachments matured in both groups. Our study showed that reattachment of tendons to cortical bone may be bettter than to cancellous bone. PMID- 12899554 TI - Cartilage injuries in distal radial fractures. AB - Subchondral hematomas have been found with arthroscopy in one third of patients with dislocated distal radial fractures. The aim of the present, prospective study was to determine whether these hematomas might cause radiographic osteoarthrosis. We studied 41 patients (age 20-57 years, 22 women) with a dislocated distal radial fracture. At the time of fracture, 12 patients had subchondral hematomas in a radiocarpal compartment without a fracture line, as defined by arthroscopy. The 1-year follow-up included clinical and radiographic examinations. At follow-up, radiographic subchondral bone plate changes occurred in unfractured compartments in 8 patients, of whom 7 had had a previous arthroscopically diagnosed subchondral hematoma (p = 0.02) in the same compartment. Of the 8 patients with radiographic changes, 4 had also developed joint space narrowing (osteoarthrosis (OA) grade 1) after 1 year and 6 after 3 years. All but 1 had had a hematoma in the same compartment. More importantly, 3 of the 16 patients with entirely extra-articular fractures had subchondral bone plate changes in a compartment corresponding to a previous subchondral hematoma (p = 0.02). One of these had also developed joint space narrowing. The patients with radiographic changes had a worse outcome, as measured with the Gartland and Werley wrist score (p = 0.06). In conclusion, subchondral hematomas in distal radial fractures can lead to early onset of mild OA and worse outcome after 1 year. PMID- 12899555 TI - Regeneration of the donor side after autogenous fibula transplantation in 53 patients: evaluation by dual x-ray absorptiometry. AB - We determined the degree of fibular regeneration at the donor site, using radiographs and dual x-ray absorptiometry, in 53 patients who underwent autogenous nonvascularized fibular transplantation for tumor reconstruction in long bones (mean follow-up 15 (3-26) years). Logistic regression was used to determine whether gender, age at transplantation, time since transplantation, bone mineral density (BMD), and length of the graft were associated with fibular regeneration. 26 patients had spontaneous complete bone regeneration. Younger age at transplant was the only predictor of fibular regeneration. In predicting fibular regeneration, sensitivity was 96% and specificity 74%, using 15 years of age as a cut-off. In the long-term follow-up, we found only gradual changes in the BMD and the values ranged from 24% to 217%. We found no correlations of bone mineral density with age, gender, length of the graft, or time since transplantation. PMID- 12899556 TI - Kinematics of the distal tibiofibular syndesmosis: radiostereometry in 11 normal ankles. AB - In 11 healthy volunteers, the normal kinematics of the tibiofibular syndesmosis of the ankle during weight bearing and external rotation stress were compared to a nonweight-bearing neutral position by radiostereometry. We found very small rotations and displacements in this "normal" group, which indicated that the fibula is closely attached to the tibia, thereby preventing larger movements at the level of the ankle. We found no common kinematic pattern during weight bearing in the neutral position. Application of a 75 Nm external rotation moment on the foot caused external rotation of the fibula between 2 and 5 degrees, medial translation between 0 and 2.5 mm and posterior displacement between 1.0 and 3.1 mm. These data can be used as normal reference values for studies of patients with suspected syndesmotic injuries. PMID- 12899557 TI - No improvement in the overall survival of 194 patients with chondrosarcoma in Finland in 1971-1990. AB - We describe the clinicopathologic profile and survival of 306 patients with chondrosarcoma reported to the Finnish Cancer Registry in 1971-1990. 218 cases were available for reevaluation. Owing to their various clinicopathologic characteristics, we excluded the histologic variants of chondrosarcoma. Therefore, the final study population included 194 patients. The minimum follow up was 9 years. The study population included 69 grade 1 tumors, 114 grade 2 tumors, and 11 grade 3 tumors. The commonest tumor sites were the chest, pelvis and femur. A local recurrence developed in 25% of the patients and metastatic lesions in 18%. The patients were treated in 31 hospitals (in 22 hospitals during the 1970s and in 26 in the 1980s), and the number of patients biopsied before the referral remained about the same from the 1970s (15%) to the 1980s (18%). The 5 and 10-year disease-specific survival rates were 70% and 57%, respectively. Multivariate analysis showed that the most important independent predictors of shortened survival were high histologic grade, age 50 years or older, and a diagnosis in the 1980s, as compared to the 1970s. Most findings accorded with reports from specialist treatment centers, but to our surprise, the survival rate declined among patients diagnosed in the 1980s versus the 1970s. The failure to improve patient survival is probably due to treatment of the patients in 31 hospitals rather than in a few centers dealing with treatment of cancer. PMID- 12899558 TI - Decreased cancer risk in patients who have been operated on with total hip and knee arthroplasty for primary osteoarthrosis: a meta-analysis of 6 Nordic cohorts with 73,000 patients. AB - 3 Nordic cohorts of total hip (THA) and total knee arthroplasty (TKA) in patients operated on for primary osteoarthrosis during 1967-1995 were partly adapted and combined for meta-analysis. THA was performed in 49,000 patients and TKA in 24,000 totaling 497,000 person years. The mean and maximum follow-up times were 6.8 and 30 years, respectively. Standardized incidence ratios (SIRs) with 95% confidence intervals (95% CI) were calculated for the observed and expected number of cancers. The expected numbers were based on national incidence rates. The overall SIRs as well as the site-specific ones were similar in the THA and TKA patients. The observed number of cancers at all sites was 7,639 and the expected one was 8,202 (SIR 0.93, 95% CI 0.91-0.95). The SIRs for lung cancer (0.69, 0.64-0.75) and laryngeal cancer (0.64, 0.44-0.92) were reduced. The incidence was also low for cancers of the stomach (SIR 0.76, 0.67-0.84), colon (SIR 0.86, 0.79-0.93) and rectum (SIR 0.89, 0.80-0.98). Among TKA patients, the only increases in SIRs were seen in cancers of the endometrium (SIR 1.36, 1.05 1.74), prostate (SIR 1.19, 1.02-1.38) and, among both THA and TKA patients, in skin melanoma (combined SIR 1.21, 1.03-1.41). PMID- 12899559 TI - Intramedullary cancellous screw fixation for nonunion of midshaft clavicular fractures. AB - 6 patients with symptomatic nonunion of midshaft clavicular fractures were treated by internal fixation with an intramedullary cancellous screw and autologous cancellous bone grafting. At follow-up after median 21 (12-72) months, all nonunions had healed. The Constant score was median 98 (57-100) points. 1 patient, with shortening of the clavicle, had poor function. PMID- 12899560 TI - HA particles can be released from well-fixed HA-coated stems. PMID- 12899561 TI - Kinetic and kinematic characteristics of gait in patients with medial knee arthrosis. PMID- 12899562 TI - Accreditation strengthens the disease management bridge over the quality chasm. PMID- 12899563 TI - Improving cancer care in a Kentucky managed care plan: a case study of cancer disease management. AB - CHA Health (CHA) is a provider-owned managed care plan serving a predominantly rural membership in Kentucky. Quality Oncology, Inc. is a national disease management company specializing in cancer care. This paper reports the results over the first 2 years of a cancer disease management program installed at CHA. The authors also review the evidence-based health services research that has guided the development and implementation of the program. The authors conclude that cancer is a disease state where specialized management can improve efficiency, treatment effectiveness, patient coordination, and outcomes reporting. PMID- 12899564 TI - The complete "how to" guide for selecting a disease management vendor. AB - Decision-makers in health plans, large medical groups, and self-insured employers face many challenges in selecting and implementing disease management programs. One strategy is the "buy" approach, utilizing one or more of the many vendors to provide disease management services for the purchasing organization. As a relatively new field, the disease management vendor landscape is continually changing, uncovering the many uncertainties about demonstrating outcomes, corporate stability, or successful business models. Given the large investment an organization may make in each disease management program (many cost 1 million dollars or more in annual fees for a moderately sized population), careful consideration must be given in selecting a disease management partner. This paper describes, in detail, the specific steps necessary and the issues to consider in achieving a successful contract with a vendor for full-service disease management. PMID- 12899565 TI - Body mass and physical capacity indicators of hip osteoarthritis patients with and without malignancy histories: implications for prevention and rehabilitation. AB - The prevalence of patients with cancer histories and types of cancers prevailing among a cohort of adults with end-stage hip osteoarthritis was established in order to determine if this group might require some form of enriched pre- and postoperative rehabilitation in view of their adverse medical history. Body weights and selected physical capacity indicators were specifically compared among hip surgical patients with and without cancer histories to specify characteristics that could direct potentially desirable and improved intervention efforts. The medical records of 1,000 hip osteoarthritis surgical candidates were scrutinized, and numbers with and without malignancy histories were recorded. Malignancy typologies and selected body mass and physical capacity indices were recorded. Specific subgroup comparisons among these variables were then made for 40 cancer survivors and an age- and gender-matched subgroup of 40 otherwise healthy osteoarthritis patients, and for selected breast, prostate, and colon cancer survivors. (1) Fourteen percent of the present patient group had a cancer history. (2) The most common malignancy noted was breast cancer, followed by prostate and then colon cancer. (3) Among subjects matched for age and gender, 85% with a cancer history were overweight or obese, compared with 60% of those with no comorbid disease history. (4) Patients with cancer histories were more impaired immediately before, and after, surgery than patients with no cancer history. (5) Patients with breast and colon cancer histories had significantly slower recovery rates after hip surgery than those with a prostate cancer history (p < 0.05). Thus, breast, prostate, and colon cancer survivors constitute a modest proportion of patients undergoing surgery for painful disabling hip osteoarthritis. As a subgroup, cancer survivors, especially breast cancer survivors, are overweight, and more impaired before and after surgery than adults of the same age without a cancer history undergoing hip surgery. PMID- 12899566 TI - The JeffCare preceptor model for asthma: a primary care physician tutorial training model. AB - The societal and economic impact of asthma is a well-documented phenomenon in this country. Despite improved knowledge and techniques of care, there have been signs of worsening morbidity/mortality and a seeming disconnect between physician and patients as regards communication of care strategies. In an attempt to fashion innovative educational strategies to enhance primary care physician (PCP) and caregiver efficiencies in improving patient outcomes (clinical and financial), the "Preceptor" model of one-to-one PCP/asthma specialist has been developed. Review of utilization and prescribing data demonstrates a clear pattern of statistically significant cost improvement in the aggregate care setting, as well as improved appropriateness of use of proper asthma medications. Use of the "Preceptor" model of PCP learning is an effective and unique way to enhance both caregiver knowledge and improved care efficiency in asthma management. PMID- 12899567 TI - The HEDIS performance NAVIGATOR for controlling high blood pressure: a resource to assist health plans improve patient adherence. AB - This article describes how the resources in the Health Plan Employer and Data Information Set (HEDIS; a registered trademark of the National Committee for Quality Assurance) Performance NAVIGATOR for Controlling High Blood Pressure (HEDIS NAVIGATOR) developed by Merck & Co., Inc. assist managed care organizations in understanding and encouraging the active involvement of their members in adhering to a treatment plan developed in partnership by physicians and patients. (HEDIS is a set of standardized performance measures that enable purchasers and consumers to make informed decisions regarding the quality of managed care organizations.) The discussion uses the Health Belief and Transtheoretical Models as a framework to better understand patient behavior change. Targeted strategies to facilitate adherence are also addressed. Examples of structured programs using nationally recommended guidelines and HEDIS NAVIGATOR resources are included to illustrate coordinated efforts to assist patients with high blood pressure adhere to their treatment plan and help managed care organizations improve patient outcomes and HEDIS scores. PMID- 12899568 TI - Vaccine adjuvants. PMID- 12899569 TI - Survey of human-use adjuvants. AB - Although there are only four adjuvants used in licensed vaccines for humans, a wealth of information on novel vaccine adjuvants has become available in both animal models and clinical studies over the past decade. Many vaccine candidates require immunopotentiation to achieve a satisfactory immune response, which is driving the search for new and safer approaches. In this review, we take a brief look at what is known of the mechanisms of action, consider some of the elements of product development, then survey several of the classes of adjuvants within the context of human trials. PMID- 12899570 TI - Virosomal adjuvanted antigen delivery systems. AB - The development of novel and increasingly safer vaccines frequently utilizes well characterized antigens, in particular highly purified proteins or synthetic peptides. In spite of some achievements, this approach is frequently impeded by the fact that such antigens are often poor immunogens when administered alone. This fact has necessitated the development of suitable adjuvants that possess the ability to enhance the immunogenicity of a given antigen, preferably with little or no side effects. This paper discusses one of the successes of vaccinology of the past decade: virosomal vaccines. The principles of the concept, immunoadjuvant action and application of virosomes in two currently licensed vaccines are detailed, with specific reference to the induction of both humoral and cellular immunity. PMID- 12899571 TI - MF59-adjuvanted vaccines: increased immunogenicity with an optimal safety profile. AB - The need to enhance the immunogenicity of purified subunit antigens has prompted the development of several new adjuvants. However, many of these new molecules have demonstrated a reactogenicity profile that is not suitable for their inclusion in vaccines for human use. In this context, the adjuvant emulsion MF59 has been developed, tested in combination with different antigens in several animal models and subsequently evaluated in humans. Clinical trials with several MF59-adjuvanted vaccines have been performed in different age groups (from newborns to the elderly) and have shown an increased immunogenicity of coadministered antigens, associated with a high level of safety and tolerability. MF59 has been the first adjuvant to be licensed for human use after alum and, as part of an enhanced influenza vaccine for the elderly, is now available in the marketplace of several countries worldwide. PMID- 12899572 TI - Mucosal adjuvants and anti-infection and anti-immunopathology vaccines based on cholera toxin, cholera toxin B subunit and CpG DNA. AB - The mucosal immune system consists of an integrated network of lymphoid cells that work in concert with innate host factors to promote host defence. Mucosal immunization can be used both to protect the mucosal surfaces against colonization and invasion by microbial pathogens and to provide a means for immunological treatment of selected autoimmune, allergic or infectious immunopathological disorders through the induction of antigen-specific tolerance. The development of mucosal vaccines, whether for prevention of infectious diseases or for oral tolerance immunotherapy, requires efficient antigen delivery and adjuvant systems. Significant progress has recently been made to generate partly or wholly detoxified derivatives of cholera toxin (including the completely nontoxic cholera toxin B subunit) and the closely related Escherichia coli heat-labile enterotoxin, with retained adjuvant activity. Cholera toxin B subunit is a protective component of a widely registered oral vaccine against cholera, and has proven to be a promising vector for either giving rise to anti infective immunity or for inducing peripheral anti-inflammatory tolerance to chemically or genetically linked foreign antigens administered mucosally. Promising advances have also recently been made in the design of efficient mucosal adjuvants based on bacterial DNA that contains CpG-motifs and various imidazoquinoline compounds binding to different Toll-like receptors on mucosal antigen-presenting cells. PMID- 12899573 TI - Enhancement of antigen-specific immunity via the TLR4 ligands MPL adjuvant and Ribi.529. AB - MPL (Corixa) adjuvant is a chemically modified derivative of lipopolysaccharide that displays greatly reduced toxicity while maintaining most of the immunostimulatory activity of lipopolysaccharide. MPL adjuvant has been used extensively in clinical trials as a component in prophylactic and therapeutic vaccines targeting infectious disease, cancer and allergies. With over 33,000 doses administered to date, MPL adjuvant has emerged as a safe and effective vaccine adjuvant. Recently, scientists at Corixa Corporation have developed a library of synthetic lipid A mimetics (aminoalkyl glucosaminide 4-phosphates) with demonstrated immunostimulatory properties. Similar to MPL adjuvant, these synthetic compounds signal through Toll-like receptor 4 to stimulate the innate immune system. One of these compounds, Ribi.529 (RC-529), has emerged as a leading adjuvant with a similar efficacy and safety profile to MPL adjuvant in both preclinical and clinical studies. PMID- 12899574 TI - Development of RTS,S/AS02: a purified subunit-based malaria vaccine candidate formulated with a novel adjuvant. AB - During the past decade, tremendous progress has been made in process development allowing for the production of large quantities of recombinant antigens, as well as in the understanding of the immune mechanisms underlying protection. Parallel to this, various and numerous adjuvant systems have been developed and tested in animal models and in clinical trials but have rarely induced protection. This review will discuss the development of a new adjuvant system (AS02) in combination with a malaria vaccine antigen candidate. To date, this vaccine is the only one to demonstrate protection in man in artificial challenge as well as in natural field trials. It has been established that this adjuvant system is capable of eliciting high antibody titers along with strong cell-mediated immunity which both contribute to the efficacy of the vaccine. PMID- 12899575 TI - Development of a leishmaniasis vaccine: the importance of MPL. AB - The parasite Leishmania has been used for pioneering work to define T-cell subsets and cytokine patterns mediating susceptibility or resistance to infectious pathogens. This understanding has been essential for the development of a new generation of candidate vaccines for major diseases, such as leishmaniases themselves, tuberculosis and others. It is clear that effective vaccines can be developed through a combination of both antigen and adjuvant selection. Until recently, no adjuvants acceptable for use in human T-cell vaccines were available. However, one such adjuvant, monophosphoryl lipid A, has been shown to be safe and effective. Just as the understanding of T-cell responses has been necessary for the development of a new generation of vaccines, an understanding of signaling by antigen-presenting cells has been essential for adjuvant selection. A combination of antigens and an adjuvant that is effective at promoting durable T-helper 1 responses and is safe for human use comprise a promising vaccine candidate, Leish-111f. This vaccine has potential application in both the prevention and treatment of leishmaniasis. PMID- 12899576 TI - Transcutaneous immunization and immunostimulant strategies: capitalizing on the immunocompetence of the skin. AB - The skin is an attractive target for vaccine delivery. Topical application of adjuvants results in potent immune responses and good safety profiles. Adjuvants can be coadministered in a patch with vaccine antigens (transcutaneous immunization) or similar delivery format, or administered separately with an injection or IS patch (Iomai), leading to enhanced immune responses. These observations have moved into the clinic, highlighting the likelihood that skin delivery of vaccines will play an important future role in vaccine applications. PMID- 12899577 TI - Microparticles as vaccine adjuvants and delivery systems. AB - Adjuvants can be broadly divided into two groups, based on their principal mechanisms of action: vaccine delivery systems and immunostimulatory adjuvants. Vaccine delivery systems are generally particulate (e.g., emulsions, microparticles, immunostimulatory complexes and liposomes) and function mainly to target associated antigens into antigen-presenting cells. However, increasingly, more complex formulations are being developed in which delivery systems are exploited both for the delivery of antigens and also for the delivery of coadministered immunostimulatory adjuvants. The rationale for this approach is to ensure that both antigen and adjuvant are delivered into the same population of antigen-presenting cells. In addition, delivery systems can focus the effect of the adjuvants onto the key cells of the immune system and limit the systemic distribution of the adjuvant, to minimize its potential to induce adverse effects. The formulation and delivery of potent adjuvants in microparticles may allow the development of prophylactic and therapeutic vaccines against cancers and chronic infectious diseases, which are currently poorly controlled. In addition, microparticle formulations may also allow vaccines to be delivered mucosally. PMID- 12899578 TI - Mutants of the Escherichia coli heat-labile enterotoxin as safe and strong adjuvants for intranasal delivery of vaccines. AB - Cholera toxin and Escherichia coli heat-labile enterotoxin are powerful mucosal adjuvants but their high toxicity hampers their use in humans. Site-directed mutagenesis has allowed the generation of several cholera toxin and E. coli heat labile enterotoxin mutants with abolished or strongly reduced toxicity that still retain strong mucosal adjuvanticity. Among them, LTK63 (Ser to Lys substitution at position 63 in the A subunit) is completely nontoxic and LTR72 (Ala to Arg at position 72) retains a very low residual enzymatic activity. Both of them have been shown to be safe and effective in enhancing the immunogenicity of intranasally coadministered vaccines, also resulting in protective responses in several animal models. Clinical grade preparations of these mutants have now been produced, tested in animals and proven to be totally safe. Indeed, they did not induce any inflammatory event in the respiratory tract nor, more importantly, in the olfactory bulbs and in the meninges. The fully nontoxic LTK63 mutant has now been successfully tested in human volunteers with a trivalent subunit influenza vaccine. PMID- 12899579 TI - Nonclinical safety evaluation of Escherichia coli heat-labile toxin mucosal adjuvant as a component of a nasal influenza vaccine. AB - Conventional influenza vaccines currently in use are administered parenterally and generally confer good protection against systemic disease through the induction of high titers of serum virus-neutralizing antibodies. Parenteral vaccines are suboptimal in that they fail to induce a local mucosal response that may prevent the early stages of virus infection. Thus, the intranasal administration of a vaccine may provide a viable alternative to the parenteral route. Indeed, intranasal administration of vaccine antigens when formulated with an appropriate mucosal adjuvant (e.g., bacterial toxins), results in a vigorous local and systemic immune response. This review discusses the nonclinical safety evaluation of Escherichia coli heat-labile toxin as a mucosal adjuvant for an intranasally administered influenza vaccine. PMID- 12899580 TI - CpG DNA as a vaccine adjuvant. AB - Synthetic oligodeoxynucleotides containing unmethylated CpG motifs act as immune adjuvants, accelerating and boosting antigen-specific antibody responses by up to 500-fold. CpG motifs promote the production of T-helper 1 and pro-inflammatory cytokines and induce the maturation/activation of professional antigen-presenting cells (including macrophages and dendritic cells). These effects are optimized by maintaining close physical contact between the CpG DNA and the immunogen. Coadministering CpG DNA with a variety of vaccines has improved protective immunity in animal challenge models. Ongoing clinical studies indicate that CpG oligodeoxynucleotides are safe and well-tolerated when administered as adjuvants to humans and in some cases increase vaccine-induced immune responses. PMID- 12899581 TI - Cytokines as clinical adjuvants: how far are we? AB - In spite of the explosive growth in the discovery of cytokines and chemokines and in the understanding of their modes of action, clinical use of such agents as adjuvants has been primarily restricted to patients with cancer or chronic viral infections suffering from various levels of immune impairment and for whom the chemotherapeutic armamentarium, as well as other forms of immunotherapy, have been exhausted. This cautious approach has been justified by the difficulties inherent to the biological function and delivery of such pleiotropic agents, where doses needed to achieve the targeted immune enhancement often result in serious side effects, especially during systemic administration. In addition, optimization of dosages, administration schedules and biological effects in humans often do not correlate well with preclinical data derived from animal models. Nevertheless, novel preventive immunization strategies that target a precise type of immune response in immunocompetent individuals are expected to greatly benefit from the incorporation of cytokines and chemokines. This review provides an overview of current clinical administration of cytokines as well as a description of select Phase I testing of new agents designed to enhance immune defenses in vivo. PMID- 12899582 TI - Vaccine adjuvant technology: from mechanistic concepts to practical applications. AB - Distinct types of immune responses are required for efficient elimination of different pathogens. Programming of the desired type of immune response by safe nonreplicating vaccines requires suitable vaccine adjuvants. Adjuvants largely determine the magnitude and quality of immune responses specific for the coadministered antigen. Unfortunately, rational vaccine design requiring a rational choice of vaccine adjuvant, is hampered by a lack of knowledge about the mechanism(s) of vaccine adjuvant activity. The current review addresses different critical immunological processes possibly explaining adjuvant functions. In addition, we discuss traditional vaccine adjuvant formulations and their possible mode of action. Finally, we reflect on the latest technologies for the identification of novel adjuvants using molecular analysis of immune activation and functional genomics. PMID- 12899583 TI - Sexual function in men older than 50 years of age: results from the health professionals follow-up study. AB - BACKGROUND: Although many studies have provided data on erectile dysfunction in specific settings, few studies have been large enough to precisely examine age specific prevalence and correlates. OBJECTIVE: To describe the association between age and several aspects of sexual functioning in men older than 50 years of age. DESIGN: Cross-sectional analysis of data from a prospective cohort study. SETTING: U.S. health professionals. PARTICIPANTS: 31 742 men, age 53 to 90 years. MEASUREMENTS: Questionnaires mailed in 2000 asked about sexual function, physical activity, body weight, smoking, marital status, medical conditions, and medications. Previous biennial questionnaires since 1986 asked about date of birth, alcohol intake, and other health information. RESULTS: When men with prostate cancer were excluded, the age-standardized prevalence of erectile dysfunction in the previous 3 months was 33%. Many aspects of sexual function (including overall function, desire, orgasm, and overall ability) decreased sharply by decade after 50 years of age. Physical activity was associated with lower risk for erectile dysfunction (multivariable relative risk, 0.7 [95% CI, 0.6 to 0.7] for >32.6 metabolic equivalent hours of exercise per week vs. 0 to 2.7 metabolic equivalent hours of exercise per week), and obesity was associated with higher risk (relative risk, 1.3 [CI, 1.2 to 1.4] for body mass index >28.7 kg/m2 vs. <23.2 kg/m2). Smoking, alcohol consumption, and television viewing time were also associated with increased prevalence of erectile dysfunction. Men who had no chronic medical conditions and engaged in healthy behaviors had the lowest prevalence. CONCLUSIONS: Several modifiable health behaviors were associated with maintenance of good erectile function, even after comorbid conditions were considered. Lifestyle factors most strongly associated with erectile dysfunction were physical activity and leanness. PMID- 12899584 TI - Effects of losartan or atenolol in hypertensive patients without clinically evident vascular disease: a substudy of the LIFE randomized trial. AB - BACKGROUND: Cardiovascular morbidity and mortality are reduced by treatment with the angiotensin II AT(1)-receptor antagonist losartan compared with conventional treatment with the beta-blocker atenolol in patients with hypertension and electrocardiogram-defined left ventricular hypertrophy, many of whom had known vascular disease. OBJECTIVE: To determine whether losartan reduces cardiovascular event rates in lower-risk hypertensive patients without clinically evident vascular disease. DESIGN: Subgroup analysis of a randomized trial. SETTING: The Losartan Intervention for Endpoint reduction in hypertension (LIFE) study. PATIENTS: 6886 men and women (57% women) 55 to 80 years of age (average, 66 years) with essential hypertension (sitting blood pressure, 160 to 200/95 to 115 mm Hg [average, 174/98 mm Hg]) and electrocardiogram-defined left ventricular hypertrophy who did not have clinically evident vascular disease. INTERVENTION: Patients were randomly assigned to once-daily double-blind treatment with losartan or atenolol. MEASUREMENTS: An end point committee ascertained end points (cardiovascular death, stroke, or myocardial infarction). RESULTS: Blood pressure was reduced similarly by losartan and atenolol. The primary composite end point occurred in 282 losartan-treated patients (17.5 per 1000 patient-years) and 355 atenolol-treated patients (21.8 per 1000 patient-years; relative risk, 0.81 [95% CI, 0.69 to 0.95]; P = 0.008). Cardiovascular death occurred in 103 losartan treated patients and 132 atenolol-treated patients (relative risk, 0.80 [CI, 0.62 to 1.04]; P = 0.092), stroke (nonfatal and fatal) occurred in 125 losartan treated patients and 193 atenolol-treated patients (relative risk, 0.66 [CI, 0.53 to 0.82]; P < 0.001), and myocardial infarction (nonfatal and fatal) occurred in 110 losartan-treated patients and 100 atenolol-treated patients (relative risk, 1.14 [CI, 0.87 to 1.49]; P > 0.2). New-onset diabetes occurred less often in patients treated with losartan (n = 173) than in patients treated with atenolol (n = 254) (relative risk, 0.69 [CI, 0.57 to 0.84]; P < 0.001). Benefits of losartan treatment were numerically smaller, but not significantly so, in patients with preexisting vascular disease. CONCLUSION: In hypertensive patients without clinically evident vascular disease, losartan was more effective than atenolol in preventing cardiovascular morbidity and death, predominantly stroke, independent of blood pressure reduction. PMID- 12899585 TI - Chlamydia trachomatis among patients infected with and treated for Neisseria gonorrhoeae in sexually transmitted disease clinics in the United States. AB - BACKGROUND: For two decades, treatment guidelines for sexually transmitted diseases (STDs) have recommended empirical co-treatment for chlamydia when patients are treated for gonorrhea. Because the epidemiology of and diagnostic testing for STDs have changed over time, co-treatment may no longer be needed as a clinical or public health strategy. OBJECTIVE: To assess the prevalence of chlamydia among patients at STD clinics who are infected with and treated for Neisseria gonorrhoeae and to determine whether co-treatment recommendations are still justified. DESIGN: Cross-sectional analysis of data from a multisite study. SETTING: Five public STD clinics (Baltimore, Maryland; Denver, Colorado; Long Beach, California; Newark, New Jersey; and San Francisco, California), July 1993 through October 1995. PATIENTS: 3885 heterosexual patients (2184 men and 1701 women) who agreed to participate in a trial of counseling interventions and had conclusive results from diagnostic tests for gonorrhea and chlamydia performed routinely as part of the trial. MEASUREMENTS: Infection with Chlamydia trachomatis as determined by polymerase chain reaction. RESULTS: Chlamydia trachomatis was detected in 20% (95% CI, 16% to 24%) of 411 men and 42% (CI, 35% to 50%) of 151 women with laboratory-confirmed N. gonorrhoeae. Chlamydia trachomatis was detected in 19% (CI, 15% to 22%) of 410 men and 35% (CI, 28% to 43%) of 154 women with treatment indications for gonorrhea who would not otherwise have been treated for chlamydia: chlamydia prevalence among these patients was significantly higher than among patients without treatment indications for either gonorrhea or chlamydia: 7% in men and 9% in women (relative risk, 2.58 [CI, 1.92 to 3.47] and 4.12 [CI, 3.05 to 5.57], respectively). CONCLUSION: The frequent presence of chlamydia among patients at STD clinics who received treatment for gonorrhea, including sex partners of gonorrhea-infected patients, supports continuing current recommendations for co treatment. PMID- 12899587 TI - Diagnosis of adrenal insufficiency. AB - BACKGROUND: The cosyntropin stimulation test is the initial endocrine evaluation of suspected primary or secondary adrenal insufficiency. PURPOSE: To critically review the utility of the cosyntropin stimulation test for evaluating adrenal insufficiency. DATA SOURCES: The MEDLINE database was searched from 1966 to 2002 for all English-language papers related to the diagnosis of adrenal insufficiency. STUDY SELECTION: Studies with fewer than 5 persons with primary or secondary adrenal insufficiency or with fewer than 10 persons as normal controls were excluded. For secondary adrenal insufficiency, only studies that stratified participants by integrated tests of adrenal function were included. DATA EXTRACTION: Summary receiver-operating characteristic (ROC) curves were generated from all studies that provided sensitivity and specificity data for 250-microg and 1-microg cosyntropin tests; these curves were then compared by using area under the curve (AUC) methods. All estimated values are given with 95% CIs. DATA SYNTHESIS: At a specificity of 95%, sensitivities were 97%, 57%, and 61% for summary ROC curves in tests for primary adrenal insufficiency (250-microg cosyntropin test), secondary adrenal insufficiency (250-microg cosyntropin test), and secondary adrenal insufficiency (1-microg cosyntropin test), respectively. The area under the curve for primary adrenal insufficiency was significantly greater than the AUC for secondary adrenal insufficiency for the high-dose cosyntropin test (P < 0.001), but AUCs for the 250-microg and 1-microg cosyntropin tests did not differ significantly (P > 0.5) for secondary adrenal insufficiency. At a specificity of 95%, summary ROC analysis for the 250-microg cosyntropin test yielded a positive likelihood ratio of 11.5 (95% CI, 8.7 to 14.2) and a negative likelihood ratio of 0.45 (CI, 0.30 to 0.60) for the diagnosis of secondary adrenal insufficiency. CONCLUSIONS: Cortisol response to cosyntropin varies considerably among healthy persons. The cosyntropin test performs well in patients with primary adrenal insufficiency, but the lower sensitivity in patients with secondary adrenal insufficiency necessitates use of tests involving stimulation of the hypothalamus if the pretest probability is sufficiently high. The operating characteristics of the 250-microg and 1-microg cosyntropin tests are similar. PMID- 12899586 TI - The effect of selective intestinal decontamination on the hyperdynamic circulatory state in cirrhosis. A randomized trial. AB - BACKGROUND: Peripheral vasodilatation is central to the pathogenesis of the accompanying hyperkinetic circulatory state and portal hypertension in cirrhotic patients. Selective intestinal decontamination with norfloxacin has been demonstrated to partially correct nitric oxide production in the forearm vasculature of cirrhotic patients. OBJECTIVE: To examine the effects of selective intestinal decontamination on regional and systemic hemodynamics in cirrhotic patients. DESIGN: Randomized, double-blind, placebo-controlled, crossover study. SETTING: Alfred Hospital, Melbourne, Australia. PATIENTS: 14 patients with alcohol-related cirrhosis and 14 matched healthy controls. INTERVENTION: Norfloxacin, 400 mg twice daily, for 4 weeks. MEASUREMENTS: Venous occlusion plethysmography was used to determine forearm blood flow. Cardiac output and the hepatic venous pressure gradient were determined after cardiac catheterization. Glomerular filtration rate was assessed by measuring inulin clearance. Serum levels of endotoxin were determined by chromogenic Limulus amebocytelysate assay. RESULTS: Norfloxacin significantly diminished serum endotoxin levels (average change, -2.14 EU/mL [95% CI, -3.6 to -0.68 EU/mL]). Derived systemic vascular resistance increased significantly with norfloxacin (2.94 units [CI, 0.74 to 5.11 units]) and was accompanied by an increase in mean arterial pressure (8.70 mm Hg [CI, 2.65 to 14.73]), a trend toward decreased cardiac output (-1.207 L/min [range, 0.05 to -2.37 L/min]), a decrease in forearm blood flow (-0.99 mL/100 mL per min [CI, -1.80 to -0.17 mL/100 mL per min]), and a trend toward reduced hepatic venous pressure gradient (-2.43 mm Hg [CI, -5.2 to 0.34 mm Hg]). Norfloxacin did not significantly alter glomerular filtration rate. CONCLUSION: Selective intestinal decontamination with norfloxacin partially reverses the hyperdynamic circulatory state in cirrhotic patients without harming splanchnic or renal hemodynamics. PMID- 12899588 TI - The thermogenic effect of thyroid hormone and its clinical implications. PMID- 12899589 TI - Pathogenesis and treatment of HIV-associated renal diseases: lessons from clinical and animal studies, molecular pathologic correlations, and genetic investigations. AB - HIV infection is associated with several renal syndromes, including acute renal failure. Chronic renal failure directly linked to HIV infection includes thrombotic microangiopathic renal diseases, immune-mediated glomerulonephritides, and HIV-associated nephropathy. A renal biopsy may be necessary for diagnosis. The development of HIV-associated nephropathy has been definitively linked to renal cellular infection, but the disease affects only a minority of patients, typically men of African descent. Therefore, factors determining disease expression in infected patients must now be emphasized. The pathogenic mechanisms involved in HIV-associated renal disease remain obscure. Genetic factors, as well as renal cellular responses, mediated by HIV proteins (including an immune activated microenvironment) capable of presenting antigen in susceptible hosts probably explain most cases. HIV-associated nephropathy has a characteristic pathologic phenotype, including glomerular, tubular, and interstitial changes, and ultrastructural findings. Infection of the glomerular epithelial cell, or podocyte, and consequent structural and biochemical changes may be pivotal in pathogenesis. The HIV-1 transgenic mouse is an important model for understanding disease pathogenesis, particularly the role of HIV proteins in mediating renal tissue injury. Rigorously controlled randomized trials have not evaluated treatment, but corticosteroids and angiotensin-converting enzyme inhibitors have been used. Highly active antiretroviral therapy seems to have decreased the incidence of end-stage renal disease related to HIV infection and, in case reports, to have improved renal functional and pathologic outcomes of HIV associated nephropathy. Outcomes in patients undergoing hemodialysis and peritoneal dialysis have improved, and current research focuses on renal transplantation for treatment of HIV-infected patients. PMID- 12899590 TI - Saving office practice. PMID- 12899591 TI - Confession. PMID- 12899593 TI - Untaken steps and untold stories. PMID- 12899594 TI - bcl-2 rearrangement in mixed cryoglobulinemia. PMID- 12899595 TI - Health care--associated bloodstream infections. PMID- 12899597 TI - Hospital-onset infections: a patient safety issue. PMID- 12899598 TI - Anti-cyclic citrullinated peptide antibodies in advanced rheumatoid arthritis. PMID- 12899599 TI - The relative safety of ephedra compared with other herbal products. PMID- 12899602 TI - Summaries for patients. Sexual function in men older than 50 years of age. PMID- 12899603 TI - Summaries for patients. Oral administration of an antibiotic (norfloxacin) may help treat the cardiac and circulatory complications of liver failure. PMID- 12899604 TI - Summaries for patients. Chlamydia infection among patients receiving treatment for gonorrhea in sexually transmitted disease clinics in the United States. PMID- 12899605 TI - Summaries for patients. Benefits of losartan in patients with hypertension and left ventricular hypertrophy but no vascular disease. PMID- 12899606 TI - Airborne dispersal of Staphylococcus aureus associated with symptomatic rhinitis allergica. PMID- 12899607 TI - Osteoporotic fractures of the distal humerus in elderly women. PMID- 12899608 TI - Angioedema as a complication of upper endoscopy. PMID- 12899609 TI - Severe neutropenia and thrombocytopenia associated with infliximab. PMID- 12899610 TI - Sir2 regulation by nicotinamide results from switching between base exchange and deacetylation chemistry. AB - Life span regulation and inhibition of gene silencing in yeast have been linked to nicotinamide effects on Sir2 enzymes. The Sir2 enzymes are NAD(+)-dependent protein deacetylases that influence gene expression by forming deacetylated proteins, nicotinamide and 2'-O-acetyl-ADPR. Nicotinamide is a base-exchange substrate as well as a biologically effective inhibitor. Characterization of the base-exchange reaction reveals that nicotinamide regulates sirtuins by switching between deacetylation and base exchange. Nicotinamide switching is quantitated for the Sir2s from Archeaglobus fulgidus (Sir2Af2), Saccharomyces cerevisiae (Sir2p), and mouse (Sir2alpha). Inhibition of deacetylation was most effective for mouse Sir2 alpha, suggesting species-dependent development of this regulatory mechanism. The Sir2s are proposed to form a relatively stable covalent intermediate between ADPR and the acetyl oxygen of the acetyllysine-protein substrate. During the lifetime of this intermediate, nicotinamide occupation of the catalytic site determines the fate of the covalent complex. Saturation of the nicotinamide site for mouse, yeast, and bacterial Sir2s causes 95, 65, and 21% of the intermediate, respectively, to return to acetylated protein. The fraction of the intermediate committed to deacetylation results from competition between the nicotinamide and the neighboring 2'-hydroxyl group at the opposite stereochemical face. Nicotinamide switching supports the previously proposed Sir2 catalytic mechanism and the existence of a 1'-O-peptidyl-ADPR.Sir2 intermediate. These findings suggest a strategy for increasing Sir2 enzyme catalytic activity in vivo by inhibition of chemical exchange but not deacetylation. PMID- 12899611 TI - The hydantoin lesions formed from oxidation of 7,8-dihydro-8-oxoguanine are potent sources of replication errors in vivo. AB - Single-stranded DNA genomes have been constructed that site-specifically contain the 7,8-dihydro-8-oxo-2'-deoxyguanine (8-oxoG) oxidation products guanidinohydantoin (Gh) and the two stable stereoisomers of spiroiminodihydantoin (Sp1 and Sp2). The circular viral genomes were transfected into wild-type AB1157 Escherichia coli, and the efficiency of lesion bypass by DNA polymerase(s) was assessed. Viral progeny were analyzed for mutation frequency and type using the recently developed restriction endonuclease and postlabeling (REAP) assay. Gh was bypassed nearly as efficiently as the parent 8-oxoG but was highly mutagenic, causing almost exclusive G --> C transversions. The stereoisomers Sp1 and Sp2 were, in comparison, much stronger blocks to DNA polymerase extension and caused a mixture of G --> T and G --> C transversions. The ratio of G --> T to G --> C mutations for each Sp lesion was dependent on the stereochemical configuration of the base. All observed mutation frequencies were at least an order of magnitude higher than those caused by 8-oxoG. Were these lesions to be formed in vivo, our data show that they are absolutely miscoding and may be refractory to repair after translesion synthesis. PMID- 12899612 TI - Electrostatic tuning of ion conductance in potassium channels. AB - Members of the K(+) channel family display remarkable conservation of sequence and structure of the ion selectivity filter, whereas the rates of K(+) turnover vary widely within the family. Here we show that channel conductance is strongly influenced by charge at the channel's intracellular mouth. Introduction of a ring of negative charges at this position in KcsA, a bacterial K(+) channel, augments the conductance in a pH-dependent manner. These results are explained by a simple electrostatic effect based on known channel structures, where the negative charges serve to alter the electrical potential at the inner mouth and, thus, to increase the local K(+) concentration. In addition, removal of the conserved negative charges at equivalent positions in a high-conductance eukaryotic K(+) channel leads to a decrease in conductance. PMID- 12899613 TI - Sequence-specific and 3'-end selective single-strand DNA binding by the Oxytricha nova telomere end binding protein alpha subunit. AB - Oxytricha nova telomere end binding protein (OnTEBP) specifically recognizes and caps single-strand (T(4)G(4))(2) telomeric DNA at the very 3'-ends of O. nova macronuclear chromosomes. The discovery of proteins homologous to the N-terminal domain of the OnTEBP alpha subunit in Euplotes crassus, Schizosaccharomyces pombe, and Homo sapiens suggests that related proteins are widely distributed in eukaryotes. Previously reported crystal structures of the ssDNA binding domain of the OnTEBP alpha subunit both uncomplexed and complexed with telomeric ssDNA have suggested specific mechanisms for sequence-specific and 3'-end selective recognition of the single-strand telomeric DNA. We now describe comparative binding studies of ssDNA recognition by the N-terminal domain of the OnTEBP alpha subunit. Addition of nucleotides to the 3'-end of the TTTTGGGG telomere repeat decreases the level of alpha binding by up to 7-fold, revealing a modest specificity for a 3'-terminus relative to an internal DNA binding site. Nucleotide substitutions at specific positions within the t(1)t(2)t(3)T(4)G(5)G(6)G(7)G(8) repeat show that base substitutions at some sites do not substantially decrease the binding affinity (<2-fold for lowercase letters), while substitutions at other sites dramatically reduce the binding affinity (>20-fold decrease for the uppercase bold letter). Comparison of the structural and binding data provides unique insights into the ways in which proteins recognize and bind single-stranded DNA. PMID- 12899614 TI - Subtype specific effects of peroxisome proliferator-activated receptor ligands on corepressor affinity. AB - Natural ligands for nuclear receptors are believed to activate gene transcription by causing dissociation of corepressors and promoting the association of coactivator proteins. Using multiple biophysical techniques, we find that peptides derived from one of the nuclear receptor interacting motifs of the corepressors nuclear receptor corepressor (NCoR) and silencing mediator of retinoid and thyroid receptors (SMRT) are able to bind the ligand binding domains (LBD) of all three PPAR (peroxisome proliferator activated receptor) subtypes. Using these peptides as tools, we find that ligands designed as selective agonists for PPAR gamma promote the association of coactivator peptides and dissociation of corepressor peptides as expected on PPAR gamma but surprisingly have varied effects on the binding of corepressor peptides to the other PPAR subtypes. In particular, some members of a class of L-tyrosine-based compounds designed as selective agonists for PPAR gamma reduce the affinity for corepressor peptides on PPAR gamma but increase the affinity for the same peptides on PPAR delta and in one case on PPAR alpha. We provide structural data that suggests that the molecular basis for these observations are variations in the ligand binding pockets of the three PPAR subtypes that are perturbed differentially by individual ligands and result in altered presentations of the overlapping coactivator/corepressor binding surfaces. PMID- 12899615 TI - Experimental evidence for a beta beta alpha-Me-finger nuclease motif to represent the active site of the caspase-activated DNase. AB - The caspase-activated DNase (CAD) is an important nuclease involved in apoptotic DNA degradation. Results of a sequence comparison of CAD proteins with beta beta alpha-Me-finger nucleases in conjunction with a mutational and chemical modification analysis suggest that CAD proteins constitute a new family of beta beta alpha-Me-finger nucleases. Nucleases of this family have widely different functions but are characterized by a common active-site fold and similar catalytic mechanisms. According to our results and comparisons with related nucleases, the active site of CAD displays features that partly resemble those of the colicin E9 and partly those of the T4 endonuclease VII active sites. We suggest that the catalytic mechanism of CAD involves a conserved histidine residue, acting as a general base, and another histidine as well as an aspartic acid residue required for cofactor binding. Our findings provide a first insight into the likely active-site structure and catalytic mechanism of a nuclease involved in the degradation of chromosomal DNA during programmed cell death. PMID- 12899616 TI - Molecular characterization of binding of calcium and carbohydrates by an early activation antigen of lymphocytes CD69. AB - CD69 is the earliest leukocyte activation antigen playing a pivotal role in cellular signaling. Here, we show that a globular C-terminal domain of CD69 belonging to C-type lectins binds calcium through Asp 171, Glu 185, and Glu 187 with K(d) approximately 54 microM. Closure of the calcium-binding site results in a conformational shift of Thr 107 and Lys 172. Interestingly, structural changes in all of these amino acids lead to the formation of high-affinity binding sites for N-acetyl-D-glucosamine. Similarly, a structural change in Glu 185 and Glu 187 contributes to a high-affinity site for N-acetyl-D-galactosamine. Site-directed mutagenesis and molecular modeling allowed us to describe the structural details of binding sites for both carbohydrates. These studies explain the importance of calcium for recognition of carbohydrates by CD69 and provide an important paradigm for the role of weak interactions in the immune system. PMID- 12899617 TI - Single disulfide and linear analogues corresponding to the carboxy-terminal segment of bovine beta-defensin-2: effects of introducing the beta-hairpin nucleating sequence d-pro-gly on antibacterial activity and Biophysical properties. AB - Mammalian defensins (alpha as well as beta forms) have a beta-hairpin structural motif spanning approximately 20 residues at the carboxy-terminal end. We have investigated the antibacterial activity and biophysical properties of synthetic peptides corresponding to the carboxy-terminal segment of bovine beta-defensin-2 (BNBD-2): VRNHVTC(1)RINRGFC(2)VPIRC(3)PGRTRQIGTC(4)FGPRIKC(5)C(6)RSW (positions of disulfide bonds are C(1)[bond]C(5), C(2)[bond]C(4), and C(3)[bond]C(6)). The parent sequence chosen was RCPGRTRQIGTIFGPRIKCRSW (P1), which spans the carboxy terminal region of BNBD-2. Since the dipeptide sequence D-Pro-Gly favors nucleation of beta-hairpin structures even in acyclic peptides, analogues of P1 with one D-Pro-Gly at the central portion and two D-Pro-Gly segments near the N- and C-terminal ends were generated. An analogue in which GP (residues 14 and 15) in P1 was switched to PG was also synthesized. It was observed that the cyclic form as well as their linear forms exhibited antibacterial activity. Circular dichroism and theoretical studies indicated that while the beta-hairpin conformation is populated, there is conformational plasticity in the cyclic and linear peptides. The mode of bacterial killing was by membrane permeabilization. The entire mammalian defensin sequence does not appear to be essential for manifestation of antibacterial activity. Hence, short peptides corresponding to the C-terminal segments of mammalian defensins could have potential as therapeutic agents. PMID- 12899618 TI - Importance of hydrophobic and polar residues in ligand binding in the family 15 carbohydrate-binding module from Cellvibrio japonicus Xyn10C. AB - Modular glycoside hydrolases that degrade the plant cell wall often contain noncatalytic carbohydrate-binding modules (CBMs) that interact with specific polysaccharides within this complex macromolecule. CBMs, by bringing the appended catalytic module into intimate and prolonged association with the substrate, increase the rate at which these enzymes are able to hydrolyze glycosidic bonds. Recently, the crystal structure of the family 15 CBM (CBM15) from Cellvibrio japonicus (formerly Pseudomonas cellulosa) Xyn10C was determined in complex with the ligand xylopentaose. In this report we have used a rational design approach, informed by the crystal structure of the CBM15-ligand complex, to probe the importance of hydrophobic stacking interactions and both direct and water mediated hydrogen bonds in the binding of this protein to xylan and xylohexaose. The data show that replacing either Trp 171 or Trp 186, which stack against xylose residues n and n + 2 in xylopentaose, with alanine abolished ligand binding. Similarly, replacing Asn 106, Gln 171, and Gln 217, which make direct hydrogen bonds with xylopentaose, with alanine greatly reduced the affinity of the protein for its saccharide ligands. By contrast, disrupting water-mediated hydrogen bonds between CBM15 and xylopentaose by introducing the mutations S108A, Q167A, Q221A, and K223A had little effect on the affinity of the protein for xylan or xylohexaose. These data indicate that CBM15 binds xylan and xylooligosaccharides via the same interactions and provide clear evidence that direct hydrogen bonds are a key determinant of affinity in a type B CBM. The generic importance of these data is discussed. PMID- 12899619 TI - Complex with a phage display-derived peptide provides insight into the function of insulin-like growth factor I. AB - The dramatic improvement in the NMR spectra of insulin-like growth factor I (IGF I) in the presence of a peptide identified from a phage display library has allowed for the first time the determination of a high-resolution solution structure for much of IGF-I. The three helices of IGF-I in this complex have an arrangement similar to that seen in high-resolution crystal structures of IGF-I and insulin, although there are differences in the conformation and precise location of helix 3. A cluster of hydrophobic and basic side chains within the turn-helix motif of the peptide contact a hydrophobic patch on helices 1 and 3 of IGF-I. The importance of this patch for tight binding was verified using alanine scanning mutagenesis of the peptide in two different phage display formats. Consistent with its antagonistic activity, the peptide binds to a region implicated by mutagenesis studies to be important for association with IGF binding proteins (IGFBPs). The ability of the peptide to also inhibit signaling has important implications for the manner in which IGF-I interacts with its receptor. Interestingly, the peptide uses the same binding site as detergent and a fragment of IGFBP-5 identified in other IGF-I complexes. The ligand-induced structural variability of helix 3 in these complexes suggests that exchange between such conformations may be the source of the dynamic nature of free IGF-I and likely has functional significance for the ability of IGF-I to recognize two signaling receptors and six binding proteins with high affinity. PMID- 12899620 TI - Structure of mammalian cytochrome P450 2C5 complexed with diclofenac at 2.1 A resolution: evidence for an induced fit model of substrate binding. AB - The structure of the anti-inflammatory drug diclofenac bound in the active site of rabbit microsomal cytochrome P450 2C5/3LVdH was determined by X-ray crystallography to 2.1 A resolution. P450 2C5/3LVdH and the related enzyme 2C5dH catalyze the 4'-hydroxylation of diclofenac with apparent K(m) values of 80 and 57 microM and k(cat) values of 13 and 16 min(-1), respectively. Spectrally determined binding constants are similar to the K(m) values. The structure indicates that the pi-electron system of the dichlorophenyl moiety faces the heme Fe with the 3'- and 4'-carbons located 4.4 and 4.7 A, respectively, from the Fe. The carboxyl moiety of the substrate is hydrogen bonded to a cluster of waters that are also hydrogen bonded to the side chains of N204, K241, S289, and D290 as well as the backbone of the protein. The proximity of the diclofenac carboxylate to the side chain of D290 together with an increased binding affinity at lower pH suggests that diclofenac is protonated when bound to the enzyme. The structure exhibits conformational changes indicative of an adaptive fit to the substrate reflecting both the hydration and size of the substrate. These results indicate how structurally diverse substrates are recognized by drug-metabolizing P450 enzymes. PMID- 12899621 TI - New lytic peptides based on the D,L-amphipathic helix motif preferentially kill tumor cells compared to normal cells. AB - Despite significant advances in cancer therapy, there is an urgent need for drugs with a new mode of action that will preferentially kill cancer cells. Several cationic antimicrobial peptides, which bind strongly to negatively charged membranes, were shown to kill cancer cells slightly better than normal cells. This was explained by a slight increase (3-9%) in the level of the negatively charged membrane phosphatidylserine (PS) in many cancer cells compared to their normal counterparts. Unfortunately, however, these peptides are inactivated by serum components. Here we synthesized and investigated the anticancer activity and the role of peptide charge, peptide structure, and phospholipid headgroup charge on the activity of a new group of diastereomeric lytic peptides (containing D- and L-forms of leucine and lysine; 15-17 amino acids long). The peptides are highly toxic to cancer cells, to a degree similar to or larger than that of mitomycin C. However, compared with mitomycin C and many native antimicrobial peptides, they are more selective for cancer cells. The peptides were investigated for (i) their binding to mono- and bilayer membranes by using the surface plasmon resonance (SPR) technique, (ii) their ability to permeate membranes by using fluorescence spectroscopy, (iii) their structure and their effect on the lipid order by using ATR-FTIR spectroscopy, and (iv) their ability to bind to cancer versus normal cells by using confocal microscopy. The data suggest that the peptides disintegrate the cell membrane in a detergent-like manner. However, in contrast to native antimicrobial peptides, the diastereomers bind and permeate similarly zwitterionic and PS-containing model membranes. Therefore, cell selectivity is probably determined mainly by improved electrostatic attraction of the peptides to acidic components on the surface of cancer cells (e.g., O-glycosylation of mucines). The simple composition of the diastereomeric peptides and their stability regarding enzymatic degradation by serum components make them excellent candidates for new chemotherapeutic drugs. PMID- 12899622 TI - Differential binding of ligands to the apolipoprotein E receptor 2. AB - Apolipoprotein E receptor 2 (apoER2) is an important participant in the Reelin signaling pathway that directs cell positioning during embryogenesis. ApoER2 is a cell surface molecule that elicits intracellular signal transduction through binding of Reelin. The structural requirements for Reelin binding to apoER2 and the receptor domains involved in this process are unclear at present. Using a series of receptor mutants, we characterized the interaction of apoER2 with Reelin and compared this interaction to that of apoER2 with the receptor associated protein (RAP), an apoER2 ligand that does not induce signaling. By surface plasmon resonance we demonstrate that apoER2 exhibits 6-fold higher affinity for Reelin than the very low density lipoprotein receptor (VLDLR), which also functions as a Reelin receptor (K(D) 0.2 nM versus K(D) 1.2 nM). Acidic amino acid residues in complement-type repeat domains 1 and 3 of apoER2 are required for Reelin binding. The same regions of the receptor are also bound by RAP with a 25-fold lower affinity (K(D) 5 nM). Whereas RAP binds to apoER2 with a 1:1 stoichiometry, experimental evidence suggests that Reelin associates with two or more receptor molecules simultaneously to achieve high-affinity interaction. This finding indicates that aggregation of apoER2 by multivalent ligands such as Reelin may be the structural basis for signal transduction. PMID- 12899623 TI - Sequence gamma 377-395(P2), but not gamma 190-202(P1), is the binding site for the alpha MI-domain of integrin alpha M beta 2 in the gamma C-domain of fibrinogen. AB - The interaction between the leukocyte integrin alpha(M)beta(2) (CD11b/CD18, Mac 1, CR3) and fibrinogen mediates the recruitment of phagocytes during the inflammatory response. Previous studies demonstrated that peptides P2 and P1, duplicating gamma 377-395 and gamma 190-202 sequences in the gamma C domain of fibrinogen, respectively, blocked the fibrinogen-binding function of alpha(M)beta(2), implicating these sequences as possible binding sites for alpha(M)beta(2). To determine the role of these sequences in integrin binding, recombinant wild-type and mutant gamma C domains were prepared, and their interactions with the alpha(M)I-domain, a ligand recognition domain within alpha(M)beta(2), were tested. Deletion of gamma 383-411 (P2-C) and gamma 377-411 produced gamma C mutants which were defective in binding to the alpha(M)I-domain. In contrast, alanine mutations of several residues in P1 did not affect alpha(M)I domain binding, and simultaneous mutations in P1 and deletion of P2 did not decrease the binding function of gamma C further. Verifying the significance of P2, inserting P2-C and the entire P2 into the homologous position of the beta C domain of fibrinogen imparted the higher alpha(M)I-domain binding ability to the chimeric proteins. To further define the molecular requirements for the P2-C activity, synthetic peptides derived from P2-C and a peptide array covering P2-C have been analyzed, and a minimal recognition motif was localized to gamma(390)NRLTIG(395). Confirming a critical role of this sequence, the cyclic peptide NRLTIG retained full activity inherent to P2-C, with Arg and Leu being important residues. Thus, these data demonstrate the essential role of the P2, but not P1, sequence for binding of gamma C by the alpha(M)I-domain and suggest that the adhesive function of P2 depends on the minimal recognition motif NRLTIG. PMID- 12899624 TI - Investigating the interfacial binding of bacterial phosphatidylinositol-specific phospholipase C. AB - The interactions of PI-PLC with nonsubstrate zwitterionic [phosphatidylcholine (PC)] and anionic [phosphatidylmethanol (PMe), phosphatidylserine, phosphatidylglycerol, and phosphatidic acid] interfaces that affect the catalytic activity of PI-PLC have been examined. PI-PLC binding is strongly coupled to vesicle curvature and is tighter at acidic pH for all of the phospholipids examined. PI-PLC binds to small unilamellar vesicles (SUVs) of anionic lipids with much higher affinity (K(d) is 0.01-0.07 microM for a site consisting of n = 100 +/- 25 lipids when analyzed with a Langmuir adsorption isotherm) than to zwitterionic PC SUVs (K(d) is 5-20 microM and n = 8 +/- 3). The binding to PC surfaces is dominated by hydrophobic interactions, while binding to anionic surfaces is dominated by electrostatic interactions. The contributions of specific cationic side chains and hydrophobic groups at the rim of the alpha beta barrel to zwitterionic and anionic vesicle binding have been assessed with mutagenesis. The results are used to explain how PC activates the enzyme for both phosphotransferase and cyclic phosphodiesterase activities. PMID- 12899625 TI - Trans stimulation provides evidence for a drug efflux carrier as the mechanism of chloroquine resistance in Plasmodium falciparum. AB - The mechanism underpinning chloroquine drug resistance in the human malarial parasite Plasmodium falciparum has remained controversial. Currently considered models to explain the resistance phenotype include acquisition of a chloroquine efflux pump, changes in intracellular chloroquine partitioning, diminished binding affinity of chloroquine to its intracellular target, heme, and changes in heme crystallization. To challenge these different models, we have investigated chloroquine accumulation under trans-stimulation conditions and in the presence and absence of glucose. We show that, in chloroquine-sensitive strains, labeled chloroquine accumulation is steadily reduced as the pre-equilibrated chloroquine concentration is raised. In the resistant cells, the extent of accumulation is, strikingly, raised at the lower levels of preloading, in comparison with resistant controls in the absence of chloroquine. The trans-stimulation effect observed in chloroquine-resistant cells is strictly energy-dependent. The data are interpreted in terms of a model in which chloroquine is bound to intracellular binding sites, not different as between sensitive and resistant cells, but where, in resistant cells, there exists an energy-dependent carrier that moves chloroquine out of this intracellular compartment. A mathematical model describing the kinetics of these processes is presented. PMID- 12899626 TI - Influence of tryptophan on lipid binding of linear amphipathic cationic antimicrobial peptides. AB - We recently demonstrated that a linear 18-residue peptide, (KIGAKI)(3)-NH(2), designed to form amphipathic beta-sheet structure when bound to lipid bilayers, possessed potent antimicrobial activity and low hemolytic activity. The ability of (KIGAKI)(3)-NH(2) to induce leakage from lipid vesicles was compared to that of the amphipathic alpha-helical peptide, (KIAGKIA)(3)-NH(2), which had equivalent antimicrobial activity. Significantly, the lytic properties of (KIGAKI)(3)-NH(2) were enhanced for mixed acidic-neutral lipid vesicles containing phosphatidylethanolamine instead of phosphatidylcholine as the neutral component, while the potency of (KIAGKIA)(3)-NH(2) was significantly reduced [Blazyk, J., et al. (2001) J. Biol. Chem. 276, 27899-27906]. In this paper, we measured the lytic properties of these peptides, as well as several fluorescent analogues containing a single tryptophan residue, by monitoring permeability changes in large unilamellar vesicles with varying lipid compositions and in Escherichia coli cells. The binding of these peptides to lipid bilayers with defined compositions was compared using surface plasmon resonance, circular dichroism, and fluorescence spectroscopy. Surprisingly large differences were observed in membrane binding properties, particularly in the case of KIGAKIKWGAKIKIGAKI-NH(2). Since all of these peptides possess the same charge and very similar mean hydrophobicities, the binding data cannot be explained merely in terms of electrostatic and/or hydrophobic interactions. In light of their equivalent antimicrobial and hemolytic potencies, some of these peptides may employ mechanisms beyond simply increasing plasma membrane permeability to exert their lethal effects. PMID- 12899627 TI - C5L2, a nonsignaling C5A binding protein. AB - C5a anaphylatoxin, a potent inflammatory mediator, is known to act through a specific G protein coupled receptor. However, some of the complex effects of C5a in vivo may not be explained solely by the deletion of the known receptor. Here, we show that an orphan receptor, identified as C5L2, is a high affinity C5a binding protein. Unlike the previously described C5aR, C5L2 is obligately uncoupled from heterotrimeric G proteins, in part by virtue of an amino acid alteration in the so-called DRY sequence at the end of the third transmembrane segment. Both human and murine C5L2 bear a leucine for arginine replacement at this site. C5L2, when transfected into several cell types, is weakly phosphorylated in transfected cells following binding of C5a but does not induce significant activation of MAP kinases, mediate calcium flux, or stimulate chemotaxis. Bone marrow cells from wild type respond robustly to C5a with induction and suppression of a number of inflammation related genes. In contrast, C5a receptor deficient mice, which bear C5L2 alone, do not respond to C5a with changes in gene transcription by microarray analyses. Biophysical properties of the C5L2, including slow ligand on and off rates, absence of internalization, and relatively high affinity for the C5a des Arg metabolite, suggest that this receptor may serve to modulate C5a biological functions in vivo. Finally, in contrast to previous reports, we find absolutely no interaction of C5L2 with other anaphylatoxins C3a and C4a. PMID- 12899628 TI - Structure-function studies of human apolipoprotein A-V: a regulator of plasma lipid homeostasis. AB - To investigate structure and function relations of a new member of the exchangeable apolipoprotein family that modulates plasma lipid levels, recombinant human apolipoprotein (apo) A-V was produced in Escherichia coli and isolated by a combination of nickel chelation affinity chromatography and reversed-phase HPLC. Antibodies directed against apoA-V were generated and employed in immunoblotting experiments. Anti-apoA-V IgG gave a strong response against recombinant apoA-V from E. coli and human apoA-V expressed in transgenic mice, but did not recognize human apoA-I or apoA-IV. In neutral-pH buffers, at concentrations of >0.1 mg/mL, isolated lipid-free apoA-V is poorly soluble. By contrast, apoA-V is soluble in 50 mM sodium citrate (pH 3.0). Far-UV circular dichroism analysis and spectral deconvolution reveal that apoA-V possesses 32% alpha-helix, 33% beta-sheet, 16% beta-turn, and 18% random coil secondary structure conformers. Temperature-induced denaturation studies gave rise to a transition midpoint of 47.1 degrees C. Upon being cooled to ambient temperature from 85 degrees C, apoA-V failed to recover all of the negative ellipticity present in unheated apoA-V. ApoA-V interacts with bilayer vesicles of dimyristoylphosphatidylcholine to form discoidal complexes with diameters in the range of 15-20 nm. However, apoA-V was a poor activator of lecithin:cholesterol acyltransferase where the activity was 8.5 +/- 1.8% of that of apoA-I. Furthermore, apoA-V failed to support enhanced efflux of cholesterol from cAMP treated J774 macrophages, although low levels of efflux were obtained from unstimulated cells. Taken together, the results demonstrate recombinant apoA-V possesses unique structural and functional characteristics, in keeping with its proposed role in the modulation of plasma lipid levels. PMID- 12899629 TI - Inhibition of the activity of SRC and Abl tyrosine protein kinases by the binding of the Wiskott-Aldrich syndrome protein. AB - The Wiscott-Aldrich syndrome protein, WASP, is an effector through which cdc42, a Rho family GTPase, regulates the actin cytoskeleton in hematopoietic cells. We have found that WASP binds readily to a number of tyrosine protein kinases including the Src kinases and the Abl kinase when the proteins are coexpressed during transient transfection. Binding inhibited the activity of each of these kinases strikingly, both in vitro and in vivo. Surprisingly, the binding was not due to an interaction between the proline-rich domain of WASP and the SH3 domain of these kinases. Rather, residues 83-93 in WASP were found to bind to the catalytic domains of the kinases. Binding did not decrease the affinity of Src kinases for either ATP or a peptide substrate noticeably. Rather, the V(max) of substrate phosphorylation was reduced by the binding of the peptide. This inhibition represents a novel form of regulation of protein kinase activity and suggests that that the isolation of small molecules that exploit this inhibitory mechanism may be possible. PMID- 12899630 TI - Yeast pol eta holds a cis-syn thymine dimer loosely in the active site during elongation opposite the 3'-T of the dimer, but tightly opposite the 5'-T. AB - Polymerase eta is a member of the Y family of DNA polymerases which is able to bypass thymine dimers efficiently and in a relatively error-free manner. To elucidate the mechanism of dimer bypass, the efficiency of dAMP and pyrene nucleotide insertion opposite the thymine dimer and its N3-methyl derivatives was determined. Pol eta inserts pyrene nucleotide with greater efficiency than dAMP opposite the 3'-T of an undimerized or dimerized T and is an effective inhibitor of DNA synthesis by pol eta. Substitution of the N3H of the 3'-T of an undimerized T or a dimerized T with a methyl group has little effect on the insertion efficiency of pyrene nucleotide but greatly inhibits the insertion of dAMP. Together, these results suggest that the error-free insertion of dAMP opposite the 3'-T of the cis-syn thymine dimer happens by way of a loosely held dimer in the active site which can be displaced from the active site by pyrene nucleotide. In contrast, pol eta cannot insert pyrene nucleotide opposite the 5' T of the dimer, whereas it can insert dAMP with efficiency comparable to that opposite the 3'-T. The inability to insert pyrene nucleotide opposite the 5'-T of the dimer is consistent with the idea that while the polymerase binds loosely to a templating nucleotide, it binds tightly to the nucleotide to its 3'-side. Overall, the results show a marked difference from similar studies on pol I family polymerases, and suggest mechanisms by which this Y family polymerase can process damaged DNA efficiently. PMID- 12899631 TI - Signal sequence cleavage and plasma membrane targeting of the retinal rod NCKX1 and cone NCKX2 Na+/Ca2+ - K+ exchangers. AB - Retinal rod and cone photoreceptors express two distinct Na(+)/Ca(2+)-K(+) exchanger (NCKX) gene products. Both the rod NCKX1 and cone NCKX2 are polytopic membrane proteins thought to contain a putative cleavable signal peptide. A cleavable signal peptide is unusual for plasma membrane proteins; moreover, predictive algorithms suggest the presence of a cleavable signal peptide for all rod NCKX1 proteins and a noncleavable signal anchor for the cone NCKX2 proteins. In this study we have placed a peptide tag at different positions of the NCKX sequence to examine whether the putative signal sequence is indeed cleaved in either NCKX1 or NCKX2 proteins expressed in heterologous systems. The signal peptide was found to be, at least in part, cleaved in dolphin rod NCKX1 and in chicken and human cone NCKX2 expressed in HEK293 cells; no signal peptide cleavage was observed for chicken rod NCKX1 despite the fact that the SignalP predictive algorithm assigned this sequence to have the highest likelihood for a cleavable signal peptide among the three NCKX sequences tested here. For the two NCKX proteins that contained a cleavable signal peptide, only cleaved NCKX protein was found in the plasma membrane of HEK293 cells. Deletion of the signal sequence in both dolphin rod NCKX1 or cone NCKX2 did not affect NCKX protein synthesis but did disrupt plasma membrane targeting as judged from abolition of NCKX function and from lack of surface biotinylation. These results are consistent with delayed signal peptide cleavage for the rod and cone NCKX proteins. PMID- 12899632 TI - An infrared reflection-absorption spectroscopy study of the secondary structure in (KL4)4K, a therapeutic agent for respiratory distress syndrome, in aqueous monolayers with phospholipids. AB - KLLLLKLLLLKLLLLKLLLLK (KL(4)) has been suggested to mimic some aspects of the pulmonary surfactant protein SP-B and has been tested clinically as a therapeutic agent for respiratory distress syndrome in premature infants [Cochrane, C. G., and Revak, S. D. (1991) Science 254, 566-568]. It is of obvious interest to understand the mechanism of KL(4) function as a guide for design of improved therapeutic agents. Attenuated total reflection (ATR) IR measurements have indicated that KL(4) is predominantly alpha-helical with a transmembrane orientation in lipid multilayers (1), a geometry quite different from the originally proposed peripheral membrane lipid interaction. However, the lipid multilayer model required for ATR may not be the best experimental paradigm to mimic the in vivo function of KL(4). In the current experiments, IR reflection absorption spectroscopy (IRRAS) was used to evaluate peptide secondary structure in monolayers at the air/water interface, the physical state that best approximates the alveolar lining. In contrast to the ATR-IR results, KL(4) (2.5-5 mol %) films with either DPPC or DPPC/DPPG (7/3 mol ratio) adopted an antiparallel beta-sheet structure at all surface pressures studied > or =5 mN/m, including pressures physiologically relevant for lung function (40-72 mN/m). In contrast, in DPPG/KL(4) films, the dominant conformation was the alpha-helix over the entire pressure range, a possible consequence of enhanced electrostatic interactions. IRRAS has thus provided unique molecular structure information and insight into KL(4)/lipid interaction in a physiologically relevant state. A structural model is proposed for the response of the peptide to surface pressure changes. PMID- 12899633 TI - Folding of a monomeric porin, OmpG, in detergent solution. AB - OmpG, a porin from E. coli, has been examined in planar lipid bilayers and in detergent solution. First, bilayer recordings were used to reinforce the evidence that the functional form of OmpG is a monomer. Both pH-dependent gating and blockade by covalent modification add support to this proposal. The findings contrast with the properties of the classical porins, which function as trimers. Second, the folding of OmpG in detergent solution was examined. A water-soluble form of OmpG was obtained by dialysis from denaturant into buffer. Incubation of water-soluble OmpG in detergent results in conversion to a form that possesses the hallmarks of a beta barrel. The folding of water-soluble OmpG in detergent was monitored by circular dichroism, protease resistance, and heat modifiability. OmpG is first transformed into an intermediate with increased beta-sheet content on the time scale of minutes at 23 degrees C. This is followed by the slow acquisition of heat modifiability and protease resistance over several hours. The formation of a beta barrel during this period was demonstrated in a double cysteine mutant by using intramolecular disulfide bond formation to report N and C terminus proximity. Finally, conditions are presented for folding OmpG with greater than 90% efficiency, thereby paving the way for structural studies. PMID- 12899634 TI - Kinetic analysis of the action of tissue transglutaminase on peptide and protein substrates. AB - Tissue transglutaminase (TGase) catalyzes transfer of gamma-acyl moieties of Gln residues in peptides or protein substrates to either water or amine nucleophiles through an acyl-enzyme intermediate formed from initial acyl-transfer to an active site Cys residue. Natural substrates for this enzyme include proteins (e.g., tau, alpha-synuclein, and huntingtin) whose TGase-promoted polymerization may be causative in neurodegenerative diseases. As part of a program to find inhibitors of TGase, we have undertaken kinetic and mechanistic studies of the enzyme from guinea pig (gpTGase) and humans (hTGase). Key findings of this study include: (i) gpTGase-catalyzed transamidation of Z-Gln-Gly by Gly-OMe proceeds essentially as described above but with the involvement of substrate inhibition by Gly-OMe. This phenomena, resulting from the binding of nucleophile to free enzyme, appears to be a common feature of TGase-catalyzed reactions. (ii) Solvent deuterium isotope effects for hydrolysis of Z-Gln-Gly by gpTGase are (D)(k(c)/K(m)) = 0.45 and (D)k(c) = 3.6. While the latter results from general catalysis of deacylation, the former originates purely from the reactant state, hydrogen fractionation factor of the active site thiol with no involvement of general catalysis of acylation. (iii) Studies of the transamidation of N,N dimethylated casein by Gly-OMe and dansyl-cadaverine suggest a complex kinetic mechanism for both enzymes that reflects contributions from four reactions: Gln hydrolysis, intramolecular transpeptidation, intermolecular transpeptidation, and transamidation by added nucleophile. PMID- 12899635 TI - The strength of dehalogenase-substrate hydrogen bonding correlates with the rate of Meisenheimer intermediate formation. AB - 4-Chlorobenzoyl-coenzyme A (4-CBA-CoA) dehalogenase catalyzes the hydrolytic dehalogenation of 4-CBA-CoA to 4-hydroxybenzoyl-CoA by using an active site aspartate as the nucleophile. Formation of the corresponding Meisenheimer complex (EMc) is followed by chloride ion expulsion which forms the arylated intermediate (EAr). This is then hydrolyzed to the product. In this paper, we explore the relationship between active site polarizing forces acting on the benzoyl carbonyl and the rate of formation of the Meisenheimer complex. The polarizing forces at the C[double bond]O group were modulated by introducing site-selected mutations (A112V, Y65D, G113A, G113S, G113N, and F64P), near the C[double bond]O binding site. Using either the substrate, 4-CBA-CoA, or the substrate analogue, 4 methylbenzoyl-CoA (4-MBA-CoA), Raman difference spectroscopy provided the position of the C[double bond]O stretching frequency (nu(C)[double bond](O)) for a total of 10 enzyme-ligand complexes. In turn, the values of the C[double bond]O frequencies could be converted to differences in effective hydrogen bonding strengths between members of the series, based on earlier model studies [Clarkson, J., Tonge, P. J., Taylor, K. L., Dunaway-Mariano, D., and Carey, P. (1997) Biochemistry 36, 10192-10199]. Catalysis in the F64P, G113A, G113S, and G113N dehalogenase mutants was very slow with k(cat) values ranging from 8 x 10( 3) to 7.6 x 10(-6) s(-1). The EAr intermediate did not accumulate to a detectable level on these enzymes during a single turnover. Catalysis in the Y65D and A112V dehalogenase mutants were almost as efficient as catalysis in wild-type dehalogenase with k(cat) values of 0.1-0.6 s(-1). In wild-type dehalogenase, 22% of the bound substrate accumulated as the EAr intermediate during a single turnover (k(obs) for EAr formation = 24 s(-(1)); in the Y65D mutant, the level of accumulation is 17% (k(obs) for EAr formation = 3 s(-1)), and in the A112V mutant, the level is 23% (k(obs) for EAr formation = 17 s(-1)). The k(obs) for EAr formation in wild-type dehalogenase and the more active dehalogenase mutants (Y65D and A112V) was taken to be an estimate of the k for EMc formation, and the k(obs) for EP formation in a single turnover was taken to be an estimate of the k for EMc formation in the severely impaired mutants (F64P, G113A, G113S, and G113N). A plot of the log k(obs) for EMc formation versus the C[double bond]O stretching frequency of bound 4-CBA-CoA (or 4-MBA-CoA) is a straight line (R(2) = 0.9584). Throughout the series, nu(C)[double bond](O) varied by 61 cm(-1), corresponding to the change in hydrogen bonding enthalpy of 67 kJ/mol. The results show that changes in polarizing forces at the benzoyl carbonyl are transmitted to the benzoyl (4) position and correlate with the rate of aromatic nucleophilic addition five chemical bonds away. Interestingly, the relationship between effective polarizing forces and reactivity seen here for dehalogenase is similar to that reported for the addition-elimination reaction involving the hydrolysis of a series of acyl serine proteases. PMID- 12899636 TI - The tetraheme cytochrome CymA is required for anaerobic respiration with dimethyl sulfoxide and nitrite in Shewanella oneidensis. AB - The tetraheme c-type cytochrome, CymA, from Shewanella oneidensis MR-1 has previously been shown to be required for respiration with Fe(III), nitrate, and fumarate [Myers, C. R., and Myers, J. M. (1997) J. Bacteriol. 179, 1143-1152]. It is located in the cytoplasmic membrane where the bulk of the protein is exposed to the periplasm, enabling it to transfer electrons to a series of redox partners. We have expressed and purified a soluble derivative of CymA (CymA(sol)) that lacks the N-terminal membrane anchor. We show here, by direct measurements of electron transfer between the purified proteins, that CymA(sol) efficiently reduces S. oneidensis fumarate reductase. This indicates that no further proteins are required for electron transfer between the quinone pool and fumarate if we assume direct reduction of CymA by quinols. By expressing CymA(sol) in a mutant lacking CymA, we have shown that this soluble form of the protein can complement the defect in fumarate respiration. We also demonstrate that CymA is essential for growth with DMSO (dimethyl sulfoxide) and for reduction of nitrite, implicating CymA in at least five different electron transfer pathways in Shewanella. PMID- 12899637 TI - Thermodynamic analysis of the structural stability of the shiga toxin B-subunit. AB - The conformational stability of Shiga toxin B-subunit (STxB), a pentameric protein from Shigella dysenteriae has been characterized by high sensitivity differential scanning calorimetry and circular dichroism spectroscopy under different solvent conditions. It is shown that the thermal folding/unfolding of STxB is a reversible process involving a highly cooperative transition between folded pentamer and unfolded monomers. The conformational stability of STxB is pH dependent and because of its pentameric nature is also concentration dependent. STxB is maximally stable in the pH range from 5 to 9 (Delta G upon unfolding is close to 13 kcal per mol of monomer at 25 degrees C), and its stability decreases both at lower and at higher pH values. The pH dependence of the Gibbs energy of stabilization between pH 2.5 and 5 is consistent with the change in the ionizable state of an average of four groups per monomer upon unfolding. Structural thermodynamic calculations show that the stabilization of the STxB pentamer is primarily due to the interactions established between monomers rather than intramonomer interactions. The folding of an isolated monomer into the conformation existing in the pentamer is unfavorable and expected to be characterized by a free-energy change upon folding in the order of 2.5 kcal mol( 1) at 25 degrees C. On the average, intersubunit interaction induced upon oligomerization of folded monomers should contribute close to -13.4 kcal per mol of monomer to bring the overall Gibbs energy to the experimentally determined value at this temperature. PMID- 12899638 TI - Hydrogen exchange-mass spectrometry analysis of beta-amyloid peptide structure. AB - beta-Amyloid peptide (A beta) is the primary protein component of senile plaques in Alzheimer's disease and is believed to be responsible for the neurodegeneration associated with the disease. A beta has proven to be toxic only when aggregated; however, the structure of the aggregated species associated with toxicity is unknown. In the present study, we use hydrogen-deuterium isotope exchange (HX)-electrospray ionization mass spectrometry (MS) along with enzymatic digestion as a tool to examine at near residue level, the changes in A beta structure associated with aggregation to a fibril form. Our results show that the structure of A beta intermediate species formed early in the course of fibrillogenesis is dependent upon solvent conditions. Additionally, the HX-MS data of peptic A beta fragments suggest that the C-terminal segment of the peptide is approximately 35% protected from exchange in fibril-containing samples, relative to monomeric A beta species prepared in DMSO/H(2)O. The N terminus (residues 1-4) is completely unprotected from exchange, and the fragment containing residues 5-19 is over 50% protected from exchange in the fibril containing samples. This work contributes to our understanding of A beta structure associated with aggregation and toxicity and further application of this approach may aid in the design of agents that intervene in the A beta aggregation processes associated with neurotoxicity. PMID- 12899639 TI - Protecting innovation in bioinformatics and in-silico biology. AB - Commercial success or failure of innovation in bioinformatics and in-silico biology requires the appropriate use of legal tools for protecting and exploiting intellectual property. These tools include patents, copyrights, trademarks, design rights, and limiting information in the form of 'trade secrets'. Potentially patentable components of bioinformatics programmes include lines of code, algorithms, data content, data structure and user interfaces. In both the US and the European Union, copyright protection is granted for software as a literary work, and most other major industrial countries have adopted similar rules. Nonetheless, the grant of software patents remains controversial and is being challenged in some countries. Current debate extends to aspects such as whether patents can claim not only the apparatus and methods but also the data signals and/or products, such as a CD-ROM, on which the programme is stored. The patentability of substances discovered using in-silico methods is a separate debate that is unlikely to be resolved in the near future. PMID- 12899640 TI - Cationic antimicrobial peptides : issues for potential clinical use. AB - Many different types of organisms use antimicrobial peptides, typically 20-40 amino acids in length, for defence against infection. Most are capable of rapidly killing a wide range of microbial cells. They have been classified according to their active structures into six extensive groups. It is not yet clear how these peptides kill bacterial cells, but it is widely believed that some cationic antimicrobial peptides kill by disrupting bacterial membranes, allowing the free exchange of intra- and extra-cellular ions. The selectivity of these peptides appears to relate to differences between the external membranes of prokaryotic and eukaryotic cells. The action of the peptides may involve the formation of 'barrel-stave' or 'torroidal' pores, the introduction of packing defects in the membrane phospholipids, or large-scale disruption of the membrane by a very dense aggregation of parallel-oriented peptide, called the 'carpet mechanism'. Antimicrobial peptides are attractive candidates for clinical development because of their selectivity, their speed of action and because bacteria may not easily develop resistance against them. Some antimicrobial peptides are already in clinical and commercial use, including ambicin (nisin), polymixin B and gramicidin S. There have been several attempts at developing peptides to make them more suitable for clinical use. For those peptides that act against bacterial membranes, it is possible to differentiate between those structural features that contribute to the specificity of initial membrane binding and those that contribute to the subsequent breach of membrane integrity. The design of novel antimicrobial peptides would necessitate the optimisation of multiple parameters, a problem that has proved difficult to solve. Potential problems to be overcome include high production costs, toxicity against eukaryotic cells, susceptibility to proteolytic degradation and the development of allergies to the peptides. Biosynthesis, using recombinant DNA techniques, could make commercial scale synthesis feasible but the peptides are usually lethal to the micro organisms used to produce them. Proteolytic degradation can be reduced by modifying the peptides to contain nonstandard amino acids, or by restricting the use of peptides to topical applications. The problem of sensitisation could be overcome by the use of our own natural antibiotics to prevent or treat infections. Despite early hopes that bacteria would not easily develop resistance to antimicrobial peptides, it is clear that some strains of bacteria already have. PMID- 12899641 TI - Quorum sensing : a novel target for the treatment of biofilm infections. AB - Present-day treatment of chronic infections is based on compounds that aim to kill or inhibit growth of bacteria. Two problems are recognised to be intrinsically associated with this approach: (i) the frequently observed development of resistance to antimicrobial compounds; and (ii) the fact that all therapeutics are considerably less effective on bacteria growing as biofilms when compared with planktonic cells. The latter point is of particular importance as evidence has accumulated over the past few years that most chronic bacterial infections involve biofilms. The discovery of bacterial communication systems (quorum sensing systems) in Gram-negative bacteria which are believed to orchestrate important temporal events during the infectious process, including the production of virulence factors and the formation of biofilms, has afforded a novel opportunity to control the activity of infecting bacteria by other means than interfering with growth. Compounds that interfere with communication systems are present in nature. Such compounds should not only specifically attenuate the production of virulence factors but should also affect biofilm formation in a manner that is unlikely to pose a selective pressure for the development of resistant mutants. PMID- 12899642 TI - Selective replicating viral vectors : potential for use in cancer gene therapy. AB - Treatment of cancer is limited by toxicity to normal tissue with standard approaches (chemotherapy, surgery and radiotherapy). The use of selective replicating viral vectors may enable the targeting of gene-modified viruses to malignant tissue without toxic effect. Studies of these vectors have demonstrated tumour-selective replication and minimal evidence of replication in normal tissue. The most advanced clinical results reported involve gene-modified adenoviral vectors. Several completed, histologically confirmed responses to local/regional injection have been induced, particularly in recurrent squamous cell carcinoma involving the head and neck region. Dose limiting toxicity above 10(13) viral particles per injection has been observed. Anti-tumour effect is demonstrable in animal models without evidence of significant toxicity when these vectors are used alone or in combination with chemotherapy, radiation therapy or as gene delivery vehicles. Preliminary clinical trials, particularly with E1B deleted adenoviruses, report evidence of clinical activity in comparison with expected historical responses. Enhancement in replication selectivity to malignant tissue is also demonstrated preclinically and clinically with an E1B deleted adenovirus utilising a prostate-specific antigen promoter. Other selective replicating viral vectors such as herpes simplex virus and vaccinia virus have also been explored clinically and suggest evidence of activity in patients with cancer. Modifications may one day enable more aggressive use of these new and exciting therapeutics as systemic gene delivery vehicles. PMID- 12899643 TI - T-cell-activation inhibitors in rheumatoid arthritis. AB - As rheumatoid arthritis (RA) is a chronic inflammatory disabling disease and a cure is not available, optimisation of therapeutic strategies is mandatory. Within recent years many new details of the inflammatory cascade(s) have been elaborated, leading to new therapeutic options such as neutralisation of tumour necrosis factor-alpha (TNFalpha). T-cell inhibition is another new approach to the treatment of RA. However, it is important to note two points: first, the role of T lymphocytes in the initiation and/or perpetuation of RA is still debated controversially. Second, there are few truly T-cell-specific agents that have proven to be effective and are established in the treatment of inflammatory disorders. Leflunomide may be considered one such agent; another in development is the fusion protein CTLA4-Ig. From a clinical perspective, studies demonstrating efficacy of these agents might represent the strongest support for a role of T cells in RA. In addition to leflunomide and CTLA4-Ig, therapeutic agents with activity against T cells, including anti-CD4 antibodies, cyclosporin, tacrolimus and T-cell receptor (TCR)-Vbeta-chain vaccination strategies, have been studied in patients with RA. Combination therapies including any of these T cell-activation inhibitors with non-T-cell-specific agents such as methotrexate, antimalarials or anti-TNFalpha biologicals may prove the most effective strategies in controlling this complex disease. PMID- 12899644 TI - Meta-analysis of basiliximab for immunoprophylaxis in renal transplantation. AB - BACKGROUND: Basiliximab is a high-affinity chimeric monoclonal antibody directed against the alpha-chain of the interleukin (IL)-2 receptor. Individual studies have shown that it is highly effective in preventing acute rejection and causes no measurable incremental toxicity. However, incorporation of basiliximab immunoprophylaxis into routine practice depends upon the demonstration of benefit across treatment regimens and quantitation of the treatment effect. METHODS: This study employed a meta-analysis to examine the clinical benefit of basiliximab. Parameter estimates were derived from four randomised prospective double-blind studies conducted in 93 renal transplant centres in 18 countries. A total of 1185 adult primary allograft recipients were randomised within the centres to receive either basiliximab 20mg intravenously on days 0 and 4 or placebo, in addition to double or triple immunosuppression consisting of cyclosporin-microemulsion (Neoral((R))The use of tradenames is for product identification purposes only and does not imply endorsement.), corticosteroids, and azathioprine or mycophenolate mofetil. Key clinical events included patient and graft survival, graft rejection and complications. Analysis was performed using a variable model; odds ratios and the numbers needed to treat (NNT) to benefit or to harm one patient were calculated for each principal outcome at 6 or 12 months post-transplant. RESULTS: Basiliximab reduced the relative risk (RR) and absolute risk (AR) of clinical and biopsy-proven acute graft rejection across all treatment regimens. The overall RR of clinical acute graft rejection was decreased by 35% in patients receiving basiliximab. AR was reduced by 15.6% (pooled incidence: 28.8% vs 44.4%, p < 0.0001), and the NNT for efficacy was six. The reduction in RR of biopsy-proven rejection was similar (32%) with an absolute risk reduction (ARR) of 11.7% (pooled incidence: 25.1% vs 36.8%, p < 0.0001) and NNT of nine over 6 months. There was a concomitant reduction in the risk of graft loss which did not reach statistical significance (p = 0.14). The RR of graft loss was reduced by 26% with an AR reduction of 2.3% (pooled incidence: 6.4% vs 8.7%) and an NNT of 42 over 6 months. The risk of death was unchanged. CONCLUSIONS: Immunoprophylaxis with basiliximab produces a significant reduction in the RR and AR of clinical and biopsy-proven acute graft rejection with a trend towards a concomitant reduction in the risk of graft loss. The magnitude of protection provided by basiliximab, the fact that it is observed across treatment regimens and the safety of this therapy are arguments for its routine use in renal transplantation. PMID- 12899645 TI - Interferon-alpha (IFN alpha) daily dose versus IFN alpha plus ribavirin for treatment-naive chronic hepatitis c patients infected by genotype 1b. AB - INTRODUCTION: Infection with hepatitis C virus genotype 1b (HCV1b) is known to be a predictive factor of poor response to both interferon-alpha (IFN alpha) alone and IFN alpha plus ribavirin combination therapy.STUDY DESIGN, PATIENTS AND METHODS: This randomised study evaluated the efficacy and safety of daily IFN alpha administration versus the combination of IFN alpha plus ribavirin in treatment-naive patients infected with chronic HCV1b. Sixty-two patients were randomised to receive either human leucocyte IFN alpha 6MU three times weekly for 12 months plus ribavirin 15 mg/kg/day for the first 6 months (group A: 29 patients), or human leucocyte IFN alpha 3MU daily for 12 months (group B: 33 patients). Response was evaluated by monitoring serum alanine aminotransferase (ALT) and HCV-RNA levels during treatment and follow-up (12 months). RESULT AND CONCLUSION: Both treatment schedules were relatively well tolerated. Normal ALT levels and negative serum HCV-RNA were observed in 16 of 29 patients (55%) of group A and in 18 of 33 patients (54.5%) of group B at the end of treatment, as well as in 10 of 29 patients (34.5%) of group A and in 12 of 33 patients (36%) of group B at the end of the follow-up. There was no significant difference between the response rates obtained with the two regimens. In naive patients with chronic HCV1b infection, the efficacy of daily administration with IFN alpha is similar to that of IFN alpha plus ribavirin administered three times a week. PMID- 12899646 TI - Urofollitropin - Ferring: FERRING hFSH, hFSH - Ferring, human follicle stimulating hormone - Ferring, Bravelle. AB - Ferring Pharmaceuticals (US) is developing a highly purified form of urofollitropin [human follicle-stimulating hormone, hFSH, FERRING hFSH, Bravelle] for use in ovarian stimulation in fertility treatments. The highly purified human derived follicle-stimulating hormone (HP-FSH), administered by either intramuscular (IM) or subcutaneous (SC) injection, was in phase III trials in the US and in May 2002 the product received US FDA approval for use in conjunction with human chorionic gonadotropin for infertility patients undergoing ovulation induction. The product will be marketed as Bravelle. Ferring Pharmaceuticals has also submitted a New Drug Application for additional indications for Bravelle in infertility treatment, which is currently under review by the US FDA. Bravelle is produced from the urine of menopausal women and has been shown to be as effective as Organon's recombinant FSH product, Follistam. Preclinical studies have shown that Ferring Pharmaceuticals' hFSH can be mixed in the same syringe as purified human menopausal gonadotropin (hMG) to reduce the number of daily injections required in ovarian stimulation protocols without altering the bioactivity of FSH or luteinising hormone. In addition, Ferring Pharmaceuticals' hFSH administered SC or IM showed positive results compared with SC Repronex in a randomised, open label study in patients undergoing in vitro fertilisation.Ferring Pharmaceuticals has teamed up with the American Infertility Association and RESOLVE: the National Infertility Association to improve access to infertility treatment in the US, with the recent launch of the 'Bravelle' HEART (Helping Expand Access to Reproductive Therapy) Programme. The programme is intended to provide urofollitropin treatment at significant cost savings to patients and their physicians. Serono is also developing a highly purified urofollitropin product. PMID- 12899647 TI - Iodine-131 Tositumomab: (131)I-anti-B1 antibody, (131)I-tositumomab, anti-CD20 murine monoclonal antibody-I-131, B1, Bexxar, (131)I-anti-B1 antibody, iodine-131 tositumomab, iodine-131 anti-B1 antibody, tositumomab. AB - Iodine-131 tositumomab [B1, Bexxar , iodine-131 anti-B1 antibody] is a murine antibody conjugated to iodine 131 that recognises and binds to the B1 (CD20) antigen which is found specifically on B lymphocytes. Iodine-131 tositumomab has a dual mechanism of action. It is capable of initiating a host immune response to those B cells to which it is attached, and it also triggers apoptosis in a significant proportion of the cells to which it binds. The product was first discovered by Coulter Corporation, Miami, in collaboration with the Dana-Farber Cancer Institute and the University of Michigan. The spin-off company Coulter Pharmaceutical, upon its formation, obtained worldwide rights to iodine-131 tositumomab. (Coulter Corporation was acquired subsequently by Beckman Instruments in October 1997. The union of the two companies produced Beckman Coulter.) In December 2000, Coulter Pharmaceutical was acquired by, and merged into, Corixa Corporation.Iodine-131 tositumomab is available for licencing in Japan. Corixa Corporation and GlaxoSmithKline signed an agreement to jointly develop and commercialise iodine-131 tositumomab. The total agreement has a potential value of up to $US132 million, plus shared profits and royalties. The two companies will jointly market the antibody in the US following regulatory approval. Corixa Corporation has announced it expects iodine-131 tositumomab to be approved in the US for non-Hodgkin's lymphoma (NHL). Under the terms of the original agreement, GlaxoSmithKline will receive exclusive marketing rights outside the US, excluding Japan. However, an amended agreement between the two companies will allow Corixa Corporation to also market the product outside the US. In February 2003, the European Commission granted iodine-131 tositumomab orphan-drug designation. Corixa Corporation and GlaxoSmithKline also intend to jointly investigate the use of the product in other indications. GlaxoSmithKline may also receive access to second generation anti-CD20 compounds under its agreement with Corixa Corporation. In May 2003, Corixa Corporation entered into an agreement that will see GlaxoSmithKline market Bexxar in Canada. Under the terms of the agreement, Corixa Corporation will manufacture and supply the product to GlaxoSmithKline, who will register, market and sell it in Canada. In October 2001, Amersham PLC, a supplier of medical equipment, announced that its Amersham Health unit had signed a marketing agreement with Corixa Corporation. The agreement allows Amersham Health to market iodine-131 tositumomab in Europe. Corixa Corporation formed agreements with Boehringer Ingelheim Pharma KG and Lonza Biologics to produce the B1 antibody and radiolabelling of the antibody has been contracted out to MDS Nordion.Iodine-131 tositumomab has received orphan drug and fast track designation for the treatment of NHL. Corixa Corporation submitted a Biologics Licence Application (BLA) to the US FDA in 1999, seeking permission to market Bexxar in the US for the treatment of relapsed or refractory, low grade or transformed low grade B-cell NHL. Following a priority review, the US FDA requested that Corixa Corporation reformat certain sections and perform additional analyses of existing data in its BLA. Corixa Corporation and GlaxoSmithKline resubmitted their BLA to the US FDA in September 2000. The BLA was subsequently accepted by the US FDA in November 2000. However, in March 2001, the US FDA requested additional information in its complete review letter to Corixa Corporation and marketing partner GlaxoSmithKline. The two companies submitted data pertaining to the chemistry, manufacturing and controls section of the BLA, and to the majority of the questions regarding the clinical section of the BLA in August 2001. Corixa Corporation and GlaxoSmithKline submitted the remainder of the response to the US FDA in September 2001, following an independent review of clinical trial data. In March 2002, Corixa Corporation received another complete review letter from the US FDA, which stated that additional clinical trials would have to be conducted in order to provide adequate evidence of the safety and clinical benefit of Bexxa. The US FDA also denied Corixa Corporation's request for accelerated approval, stating that the data provided was inadequate to show that Bexxar filled an unmet medical need. Corixa Corporation has met formally with the US FDA but the two were unable to resolve their differences. Corixa Corporation will now file a formal request for dispute resolution under the Food and Drug Administration Modernisation Act. Corixa Corporation also requested a presentation of Bexxa data to the US FDA's scientific advisors. In June 2002, the US FDA granted the company's appeal for additional regulatory review. In December 2002, the US FDA's Oncologic Drugs Advisory Committee agreed that Bexxar has clinical benefit for patients with NHL. In May 2003, Corixa Corporation and GlaxoSmithKline announced that they had fulfilled many of the steps required for US FDA approval, however the US FDA has extended its review of the application for another 3 months. This extension will allow for further refinement of post marketing commitments and package insert language, and to ensure they are consistent with an updated safety database requested by the US FDA and submitted by Corixa Corporation in early April. GlaxoSmithKline was waiting for the outcome of the situation before deciding on marketing plans for Bexxar. Corixa Corporation and GlaxoSmithKline will conduct a head-to-head study of Bexxar and Idec's Zevalin, planned for mid-2003. The trial will likely be one of three phase IV studies that the US FDA requires for accelerated approval of Bexxar. Corixa Corporation initiated its Expanded Access Program for Bexxar in response to requests from physicians and patients for continued access to Bexxar during the period prior to potential US FDA marketing approval.A phase II multicentre trial of Bexxar in combination with CHOP chemotherapy is underway in the US as first-line therapy in patients with intermediate-grade NHL. Corixa Corporation has initiated a phase II trial of iodine-131 tositumomab in combination with cyclophosphamide, vincristine and prednisone for the treatment of previously untreated low-grade NHL. The trial was initiated while the company was preparing its BLA for Bexxar for use as a single agent for relapsed or refractory NHL. Corixa Corporation intends to pursue additional trials to expand the potential use of iodine-131 tositumomab to other indications, including chronic lymphocytic leukaemia. The agent is also in a clinical trial for preparation in autologous bone marrow transplant patients. The trial is designed to test the combination of iodine-131 tositumomab and chemotherapy. The trial began in 1995 and has so far enrolled 40 patients. In addition, a phase II dose-escalation trial has begun at the University of Nebraska for the combined use of iodine-131 tositumomab and chemotherapy as preparation for autologous bone marrow transplant. Corixa Corporation has received an issued US patent covering methods for administering and dosing radioimmunotherapy for the treatment of B-cell lymphomas. The patent covers iodine-131 tositumomab and other anti-CD20 antibodies used to aid in selective tumour targeting. Corixa Corporation has exclusive rights to the patent.A February 2000 media release from GlaxoSmithKline and Corixa Corporation stated that they had been issued a composition patent relating to radiolabelled monoclonal antibodies (including Bexxar) for the treatment of B-cell lymphomas. On 11 September 2001, IDEC announced that it had filed two separate lawsuits. The first lawsuit is against Corixa Corporation and the University of Michigan on six patents pertaining to products and processes related to radioimmunotherapy. They seek a declaration that Zevalin does not infringe Corixa Corporation's issued US patents. The second lawsuit involves two patents relating to cell culture media, and is against GlaxoSmithKline. IDEC's lawsuit in this case, seeks a declaration that its manufacture of Zevalin does not infringe GlaxoSmithKline's issued US patents. Corixa Corporation and GlaxoSmithKline have also filed a complaint for patent infringement against IDEC. These actions however, should have no effect on the regulatory process that Zevalin is completing, or prevent IDEC from launching the drug before iodine-131 tositumomab.A year earlier, in March 2001, the Financial Times reported that Bexxar could reach peak sales of $US120 million. In 1998, Coulter Pharmaceutical received a licencee fee payment of $US34 million from SmithKline Beecham (now GSK) in the fourth quarter of the year, as part of the joint development and commercialisation agreement for Bexxar. PMID- 12899648 TI - Human haemoglobin - Northfield. AB - PolyHeme is a chemically modified haemoglobin solution derived from human blood. It has been developed by Northfield Laboratories as an alternative to transfused blood for the treatment of acute blood loss (such as in trauma or surgery). Single molecules of haemoglobin are toxic to the kidneys and cause vasoconstriction. The haemoglobin molecules in PolyHeme are polymerised to create small chains of linked tetramers. In August 2001, Northfield Laboratories stated that it had submitted a Biologics Licence Application (BLA) to the US FDA for approval of PolyHeme as a blood substitute for use in the treatment of urgent, life-threatening blood loss. There were some undisclosed problems with the BLA, and the US FDA asked for more information before it would accept the application or allow the company to conduct a trial for this indication. In March 2003, Northfield Laboratories announced that it had received clearance from the US FDA to proceed with a pivotal phase III trial evaluating the safety and efficacy of PolyHeme in improving patient survival when used to treat severely injured and bleeding trauma patients at the scene of injury and during ambulance transport to the hospital. Because of the severity of patients' injuries, this trial will be conducted under federal regulations that allow clinical research in emergency settings without obtaining informed consent. This trial will be conducted in urban settings where transit times to hospital may be short and not long enough to demonstrate superiority to the control group, therefore dual primary endpoints of superiority and non-inferiority will be evaluated. Northfield Laboratories has initiated a public disclosure and community consultation process; because informed consent will not be possible, this is one of the US FDA requirements that must be met prior to initiating the trial. In June 2003, Northfield Laboratories reached an agreement with the US FDA on Special Protocol Assessment (SPA) for the trial. The SPA is an acknowledgment and agreement between the trial sponsor and the US FDA for successful completion of the proposed trial to form the primary basis for an efficacy claim. In October 2002, Northfield Laboratories announced that it was collaborating with the US Army in developing a treatment Investigational New Drug Application (IND) for the battlefield use of PolyHeme. The IND is being developed to gain authorisation for the use of PolyHeme in patients in shock in remote battlefield settings, where red blood cells are not available. Phase III trials in the UK, Germany and Sweden are underway with Pharmacia Corporation (now Pfizer), who has licenced PolyHeme in these countries. On 16 April 2003, Pharmacia Corporation was acquired by, and merged into, Pfizer. Northfield Laboratories' US phase III trials have focused on elective surgery indications while the European trials are focusing on surgery with acute trauma patients where large amounts of blood are lost. Up to 20 units of PolyHeme have been infused in US trials in trauma patients. This volume represents 200% of an average person's blood volume. Northfield Laboratories has estimated that of the approximately 14 million units of blood which were used for transfusions in the US in 2001, about 8.5 million units were for acute blood loss, for which patient charges amounted to an estimated $US2.5 billion. The US transfusion market consists of two principal segments: the acute blood loss segment, which accounts for about 60% of the total market according to Northfield Laboratories, and the chronic blood loss segment, accounting for the remaining 40%. The former includes transfusions in connection with trauma, surgery and unexpected blood loss, while the latter includes transfusions used in general medical applications and chronic anaemias. Northfield Laboratories believes that PolyHeme's largest potential is in the elective surgery market, at the time estimated to use approximately 6.5 million units of blood in the US annually.Earlier, on 7 March 2003, the newswires quoted er, on 7 March 2003, the newswires quoted the CEO of Northfield Laboratories, stating that it is to administer 10 000 units of PolyHeme to 20 US trauma centres, following US FDA approval of a third phase III trial. Pending market approval of PolyHeme, the company will initially produce 75 000 units of the product, but expects this to increase to 400 000 units annually. Wall Street analysts have estimated the market potential for a substitute blood product to be in the "multibillion-dollar" range, according to the CEO. However, launch of PolyHeme was at the time said to be "a couple of years away". PMID- 12899649 TI - Testosterone-topical fortigel - cellegy: fortigel, tostrex. AB - Cellegy Pharmaceuticals in the US is developing a topical gel formulation of testosterone replacement therapy [Tostrex, Fortigel] for men, based on the company's proprietary Permeate technology. The new gel formulation, containing 2% testosterone, is intended to be a more convenient alternative to injectable testosterone, and is designed to reduce the skin irritation associated with currently available dermal patch products. Fortigel (previously known as Tostrex Gel) will be provided in a proprietary metered dose canister. Fortigel is engineered to deliver physiological levels of testosterone in an easy-to-use, non greasy, non-staining topical formulation, which is designed for discreet application to the inner thigh.A similar product is also being developed for women. In April 2003, following a request by the US FDA, Cellegy changed the trade name of its topical gel formulation of testosterone from Tostrex Gel to Fortigel. The name change was made in response to the US FDA's observation that the original name could potentially create confusion on the part of medical professionals and consumers when compared with currently marketed products. Cellegy have submitted a European Marketing Authorization Application (MAA) for Fortigel as a treatment for male hormone replacement therapy or hypogonadism. Sweden is acting as the reference member state for the Mutual Recognition Procedure. Data presented at the 98th Annual Meeting of the American Urological Association have shown that the topical testosterone gel restored serum testosterone levels to within the physiological range in 92% of hypogonadal men who completed the trial. Cellegy is considering outlicencing possibilities for the product outside the US. Fortigel is covered by an issued patent in the US and patents are pending in other markets. In April 2001, Cellegy estimated the US market size for products such as Tostrex to be $US5 billion; approximately 5 million men are affected by male hypogonadism in the US. PMID- 12899650 TI - Intrauterine undernutrition and programming as a new risk of cardiovascular disease in later life. AB - It is believed that atherogenesis is a multifactorial process, which could already start in utero. Development of atherosclerosis progresses over decades and leads to the cardiovascular morbidity and mortality in adulthood. At present, we have no exact explanation for all the risk factors acting in the pathogenesis of atherosclerosis. This review should provide an overview about the possible role of intrauterine undernutrition in the development of risk factors for cardiovascular disease. Intrauterine undernutrition leads to changes in fetal growth and metabolism and programs later development of some of these risk factors. A number of experimental and human studies indicates that hypertension as well as impaired cholesterol and glucose metabolism are affected by intrauterine growth. Intrauterine undernutrition plays an important role and acts synergistically with numerous genetic and environmental factors in the development of atherosclerosis. There is evidence that undernutrition of the fetus has permanent effects on the health status of human individuals. PMID- 12899651 TI - Dehydroepiandrosterone - is the fountain of youth drying out? AB - Dehydroepiandrosterone (DHEA) and its sulphate-bound form (DHEAS) are important steroids mainly of adrenal origin. Their physiological and pathophysiological functions are not yet fully identified, although a number of various possible features have been hypothesized. Most popular is the description of the "hormone of youth" as the long-term dynamics of DHEA levels are characterized by a sharp age-related decline in the late adulthood and later. Low levels of DHEA are, however, associated not only with the ageing process but also with diabetes mellitus, cardiovascular diseases and some neurological or immunological entities. In the past decade, a number of brief studies have concentrated on these relationships and also on the role of exogenous DHEA in health, disease and human well-being. This article tries to summarize some of the most important facts achieved recently. PMID- 12899652 TI - Preconditioning by hypoventilation increases ventricular arrhythmia threshold in Wistar rats. AB - Hypoventilation, as one of ventilatory disorders, decreases the electrical stability of the heart similarly as ischemia. If preconditioning by short cycles of ischemia has a cardioprotective effect against harmful influences of a prolonged ischemic period, then preconditioning by hypoventilation (HPC) can also have a similar effect. Anesthetized rats (ketamine 100 mg/kg + xylasine 15 mg/kg i.m., open chest experiments) were subjected to 20 min of hypoventilation followed by 20 min of reoxygenation (control group). The preconditioning (PC) was induced by one (1PC), two (2PC) or three (3PC) cycles of 5-min hypoventilation followed by 5-min reoxygenation. The electrical stability of the heart was measured by a ventricular arrhythmia threshold (VAT) tested by electrical stimulation of the right ventricle. Twenty-minute hypoventilation significantly decreased the VAT in the control and 1PC groups (p<0.05) and non-significantly in 2PC vs. the initial values. Reoxygenation reversed the VAT values to the initial level only in the control group. In 3PC, the VAT was increased from 2.32+/-0.69 mA to 4.25+/-1.31 mA. during hypoventilation (p<0.001) and to 4.37+/-1.99 mA during reoxygenation (p<0.001). It is concluded that cardioprotection against the hypoventilation/ reoxygenation-induced decrease of VAT proved to be effective only after three cycles of HPC. PMID- 12899653 TI - Time course of leukocyte response and free radical release in an early reperfusion injury of the superior mesenteric artery. AB - The sequence of changes in circulating immune cells and in free radical production was studied during the small intestine reperfusion. Rat small intestine ischemia/reperfusion was induced by a 45 min superior mesenteric artery occlusion followed by a 4-hour reperfusion. Samples of peripheral blood were collected every 20 min during reperfusion. While the number of polymorphonuclear leukocytes increased significantly both in the sham-operated controls and the experimental group (about 400 per cent at the end of reperfusion), a decrease in lymphocyte counts to 60 per cent was observed in the experimental group only. Although there were no changes in the counts of total T lymphocytes, a significant reduction in B cell counts was observed. Flow-cytometrical measurements showed no changes in the Tc subpopulation, while the Th subpopulation increased in the experimental group only. Free radical generation in blood (luminometric measurements) increased gradually and reached an eight fold level by the end of reperfusion in both groups. Thus, it has been shown that the increase in free radical production is mainly due to the increased number of polymorphonuclear leukocytes mobilized already at the initial stages of reperfusion. The reduction in B lymphocyte population is probably due to homing mechanisms PMID- 12899654 TI - Red wine polyphenols induce vasorelaxation by increased nitric oxide bioactivity. AB - The aim of the present study was to investigate the mechanism of vasorelaxant responses induced by red wine polyphenolic compounds (Provinol). Rings of rat femoral artery with or without functional endothelium were set up in a myograph for isometric recording and precontracted with phenylephrine (10(-5) M). Provinol in cumulative doses (10(-9) to 10(-3) mg/ml) elicited endothelium- and dose dependent relaxation of the artery with maximal relaxation of 56 per cent at the concentration of 10(-5) mg/ml. The relaxant responses to Provinol correlated well with the increase of NO synthase activity in the vascular tissue after administration of cumulative doses of Provinol (10(-9) to 10(-3) mg/ml). N(G) nitro-L-arginine methylester (L-NAME, 3x10(-4) M) significantly attenuated the endothelium-dependent relaxation produced by Provinol. Administration of L arginine (3x10(-5) M) restored the relaxation inhibited by L-NAME. The relaxant responses of Provinol were abolished in the presence of Ca(2+)-entry blocker, verapamil (10(-6) M). Administration of hydrogen peroxide (H(2)O(2)) abolished acetylcholine (10(-5) M)-induced relaxation of the rat femoral artery, while administration of Provinol (10(2) mg/ml) together with H(2)O(2) helped to maintain the acetylcholine-induced relaxation. Provinol only partially affected the concentration-response curve for the NO donor sodium nitroprusside-induced relaxation in rings without endothelium. In conclusion, Provinol elicited endothelium-dependent relaxation of rat femoral artery by the Ca(2+)-induced increase of NO synthase activity and by protecting NO from degradation. PMID- 12899655 TI - Role of endothelin and nitric oxide in the pathogenesis of arterial hypertension in autosomal dominant polycystic kidney disease. AB - The pathogenesis of arterial hypertension in autosomal dominant polycystic kidney disease (ADPKD) is complex and likely dependent on interaction of hemodynamic, endocrine and neurogenic factors. We decided to evaluate the role of endothelin (ET1) and nitric oxide (NO) in the regulation of arterial blood pressure (BP) and to determine plasma levels of ET1 and NO in the group of patients with ADPKD. The ADPKD group (18 patients, 6 men + 12 women, mean age 44.6+/-11.7 years, with creatinine clearancecorrig > 1.1 ml/s) was compared with a control group of 27 healthy volunteers of comparable age. Plasma levels of ET1 assessed by direct RIA determination in the group of ADPKD patients (11.03+/-1.8 fmol/ml) were significantly increased (p<0.001) in comparison with the control group (2.66+/ 0.58 fmol/ml), while no significant differences were observed between normotensive and hypertensive patients in the ADPKD group. Serum levels of NO were evaluated according to the determination of serum levels of their metabolites - nitrites/nitrates. Serum levels of NO in the group of ADPKD patients (39.85+/-.38 micro mol/l) were significantly higher (p<0.05) in comparison with the control group (22.7+/-1.20 micro mol/l), whereas in the ADPKD group no significant differences were observed between normotensive and hypertensive patients. Thus, our study supports the concept of complex alteration of both vasoconstrictor and vasodilator systems in the pathogenesis of arterial hypertension in ADPKD. PMID- 12899656 TI - Microvascular reactivity in patients with hypercholesterolemia: effect of lipid lowering treatment. AB - Impaired NO-dependent vasodilation of resistance vessels is an early marker of an increased risk of atherosclerosis; utility of the examination of microcirculation, however, is far less established. We have therefore tested the hypothesis that hypercholesterolemia is associated with an impaired microvascular reactivity and that this defect is at least partially reversible by lipid lowering treatment. Twenty-seven otherwise healthy patients with severe hypercholesterolemia (HLP) were examined at rest and then after 10 weeks of atorvastatin treatment (20 mg/day). Skin microvascular reactivity (MVR) was examined by laser-Doppler flowmetry. Baseline MVR values of the studied group were compared to healthy control subjects, HLP patients with coronary artery disease (CAD) and diabetic patients with and without diabetic retinopathy. MVR was normal in HLP subjects without CAD. On the contrary, MVR was impaired in HLP patients with CAD. There was no effect of atorvastatin on MVR, despite the profound reduction of serum lipids. MVR values did not correlate with cholesterol levels. In diabetic subjects, the MVR was substantially impaired only in patients with retinopathy. In the subjects without retinopathy, MVR was either normal (type I diabetes) or moderately impaired (type II diabetes). MVR was thus normal in HLP patients without manifest vascular disease and was not influenced by lipid lowering therapy. Impairment in the MVR was only evident in subjects with HLP and severe CAD. These results suggest that microcirculation is not involved in the early vascular dysfunction induced by HLP and that MVR rather reflects changes which appear later in the course of the atherosclerotic disease. PMID- 12899657 TI - Involvement of phospholipids in the mechanism of insulin action in HEPG2 cells. AB - The mechanism of action by which insulin increases phosphatidic acid (PA) and diacylglycerol (DAG) levels was investigated in cultured hepatoma cells (HEPG2). Insulin stimulated phosphatidylcholine (PC) and phosphatidyl-inositol (PI) degradation through the activation of specific phospholipases C (PLC). The DAG increase appears to be biphasic. The early DAG production seems to be due to PI breakdown, probably through phosphatidyl-inositol-3-kinase (PI3K) involvement, whereas the delayed DAG increase is derived directly from the PC-PLC activity. The absence of phospholipase D (PLD) involvement was confirmed by the lack of PC derived phosphatidylethanol production. Experiments performed in the presence of R59022, an inhibitor of DAG-kinase, indicated that PA release is the result of the DAG-kinase activity on the DAG produced in the early phase of insulin action. PMID- 12899658 TI - Cerebrolysin inhibits lipid peroxidation induced by insulin hypoglycemia in the brain and heart of mice. AB - As a consequence of enhanced production of oxygen free radicals, lipid peroxidation leads to the degradation of membrane lipids and disturbances of membrane permeability. Lipid peroxidation increases under stress conditions such as hypoxia, ischemia or acidosis as well as in metabolic diseases, e.g. diabetes mellitus. We have shown that subcomatous doses of insulin (6.0 IU/kg) significantly increase thiobarbituric acid reactive substances (TBARs), especially malondialdehyde (MDA) - the endproduct of lipid peroxidation, in the brain and heart of mice. In our model of insulin-induced hypoglycemia, mice were treated with the neuroprotective, peptide-containing drug Cerebrolysin (100 mg/kg b.w.). Animals were sacrificed by decapitation two or three hours after the injection of tested substance and samples were taken to determine several serum parameters (glucose, total protein, triglycerides and lactic acid) and TBARs in the brain and heart. Although Cerebrolysin was not able to affect serum parameters after subcomatous insulin injection, the drug significantly influenced lipid peroxidation. A single injection of Cerebrolysin already decreased TBARs levels in the brain and heart tissue. Presuming that an increase of TBARs reflects disturbances of the cell membrane, we have documented a promising effect of Cerebrolysin on cell integrity. PMID- 12899659 TI - Hepatoprotective effect of rooibos tea (Aspalathus linearis) on CCl4-induced liver damage in rats. AB - Hepatoprotective properties of rooibos tea (Aspalathus linearis) were investigated in a rat model of liver injury induced by carbon tetrachloride (CCl(4)). Rooibos tea, like N-acetyl-L-cysteine which was used for the comparison, showed histological regression of steatosis and cirrhosis in the liver tissue with a significant inhibition of the increase of liver tissue concentrations of malondialdehyde, triacylglycerols and cholesterol. Simultaneously, rooibos tea significantly suppressed mainly the increase in plasma activities of aminotransferases (ALT, AST), alkaline phosphatase and billirubin concentrations, which are considered as markers of liver functional state. The antifibrotic effect in the experimental model of hepatic cirrhosis of rats suggests the use of rooibos tea as a plant hepatoprotector in the diet of patients with hepatopathies. PMID- 12899661 TI - Long release latencies are increased by acetylcholine at frog endplate. AB - Uni-quantal endplate currents (EPCs) were recorded extracellularly at the frog neuromuscular synapse and their latency dispersions expressed as P(90) were estimated in the presence of acetylcholine. Stimulation-evoked EPCs with long release latencies increased in number when acetylcholine was applied. P90, which is designated as the interval between the minimal synaptic delay and the time at which 90 per cent of all measured uni-quantal EPCs had occurred, was significantly and reversibly increased by 66 per cent from 0.51 ms to 0.85 ms in the presence of 5x10(-4) M acetylcholine. This indicates that the evoked release pattern is less synchronous and the increased asynchrony leads to a substantial drop (by 28 per cent) in the amplitude of reconstructed multi-quantal currents. PMID- 12899660 TI - In vitro glycosidation potential towards olomoucine-type cyclin-dependent kinase inhibitors in rodent and primate microsomes. AB - Interspecies differences in glycosidation potential in mammalian tissues represent a factor contributing to ambiguity when endobiotic and/or xenobiotic metabolic pathways are extrapolated from animals to man. Using the TLC/autoradiographic technique, we conducted an in vitro investigation involving mouse, rat, monkey, as well as human liver and kidney microsomes to evaluate their glycoconjugation potential towards (3)H-labeled, purine-derived selective inhibitors of cyclin-dependent kinases such as olomoucine, bohemine, roscovitine, 6-(2-hydroxybenzyl)amino-2-(1-hydroxymethyl-2-methylpropyl)amino-9 isopropylpurine (compound A-4), and 6-(3-hydroxybenzyl)amino-2-[(1(R/S) hydroxymethyl)propyl]amino-9-isopropylpurine (compound A-5) as aglycones. Principally, this study confirmed the aliphatic hydroxyl group of olomoucine-type inhibitors as a relatively suitable target for glucuronide, glucoside, xyloside, galactoside, and/or N-acetylaminoglucoside conjugation. Of the tissues examined, only the mouse microsomes were able to perform glucosidation and galactosidation reactions with the aglycones. On the other hand, monkey microsomes were superior to the mouse microsomes in a variety of glucuronide conjugates produced with compounds A-4 and A-5. PMID- 12899662 TI - Cauda equina syndrome and nitric oxide synthase immunoreactivity in the spinal cord of the dog. AB - The development of the cauda equina syndrome in the dog and the involvement of spinal nitric oxide synthase immunoreactivity (NOS-IR) and catalytic nitric oxide synthase (cNOS) activity were studied in a pain model caused by multiple cauda equina constrictions. Increased NOS-IR was found two days post-constriction in neurons of the deep dorsal horn and in large, mostly bipolar neurons located in the internal basal nucleus of Cajal seen along the medial border of the dorsal horn. Concomitantly, NOS-IR was detected in small neurons close to the medioventral border of the ventral horn. High NOS-IR appeared in a dense sacral vascular body close to the Lissauer tract in S1-S3 segments. Somatic and fiber like NOS-IR appeared at five days post-constriction in the Lissauer tract and in the lateral and medial collateral pathways arising from the Lissauer tract. Both pathways were accompanied by a dense punctate NOS immunopositive staining. Simultaneously, the internal basal nucleus of Cajal and neuropil of this nucleus exhibited high NOS-IR. A significant decrease in the number of small NOS immunoreactive somata was noted in laminae I-II of L6-S2 segments at five days post-constriction while, at the same time, the number of NOS immunoreactive neurons located in laminae VIII and IX was significantly increased. Moreover, high immunopositivity in the sacral vascular body persisted along with a highly expressed NOS-IR staining of vessels supplying the dorsal sacral gray commissure and dorsal horn in S1-S3 segments. cNOS activity, based on a radioassay of compartmentalized gray and white matter regions of lower lumbar segments and non compartmentalized gray and white matter of S1-S3 segments, proved to be highly variable for both post-constriction periods. PMID- 12899663 TI - Electroencephalogram is activated by addition of pentobarbital in the isolated perfused head of the rat. AB - To evaluate the direct effects of a barbiturate on cerebral functions without its influence on brain perfusion pressure, circulatory hormones and metabolites, the electroencephalogram (EEG) was studied in the isolated rat head. Male Wistar rats were anesthetized, and EEG electrodes were inserted into the cranium. A Krebs Ringer bicarbonate buffer solution containing heparinized rat whole blood, 20 mmol/l glucose, 200 mmol/l mannitol and 0.1 mg/ml dexamethasone was used for the perfusate. The bilateral common carotid arteries were cannulated, pumped at a rate of 6 ml/min and the head was isolated. The venous effluent was reoxygenated and recirculated into the brain. Twenty-five min after isolation of the heads pentobarbital was added to the perfusate at concentrations of 0.1, 0.5 and 2.5 mg/ml. EEG was recorded before and during perfusion. EEG activity could be recorded for more than 25 min after the beginning of perfusion. EEG activity gradually declined from 42+/-5 microV before perfusion (in vivo) to 4+/-1 microV at 25 min after the beginning of perfusion. Then, 3 min after the addition of pentobarbital, the EEG activity became significantly higher in the high dose groups; 12+/-3 microV in the 0.5 mg/ml group (p<0.05) and 12+/-1 microV in 2.5 mg/ml group (p<0.05) compared with the group without pentobarbital (2+/-2 microV). The present study suggests that a barbiturate has mitigating effects on the brain damage induced by the in vitro brain perfusion. PMID- 12899664 TI - Does the treatment of primary hyperaldosteronism influence glucose tolerance? AB - Primary hyperaldosteronism (PH) is frequently considered to be a secondary form of diabetes mellitus (DM). In our previous study we attempted to evaluate the prevalence of DM among patients with PH compared to control subjects with essential hypertension (EH). We have noted a relatively high prevalence of DM and impaired glucose tolerance in PH, but the differences between the PH and EH groups did not reach statistical significance. We performed this study to assess whether the effective treatment of PH (surgical and conservative) would improve the glucose tolerance. We have studied 24 patients with PH of the following two subtypes: aldosterone-producing adenoma (APA) treated with adrenalectomy and idiopathic hyperaldosteronism (IHA) treated with spironolactone. No significant changes of glucose levels were found in the 60th and 120th min of the oral glucose tolerance test (OGTT) in the APA group. On the other hand, fasting glucose levels were decreased significantly after adrenalectomy. Plasma glucose levels were significantly increased in the 60th min, but no differences were found in fasting values and in the 120th min in the IHA group. There was a significantly higher incidence of impaired glucose tolerance (36 per cent before, 45 per cent after treatment) and DM (9 per cent, 18 per cent) in the IHA group compared to the APA group (8 per cent, 32 per cent; DM 0 per cent, 0 per cent). In conclusion, the treatment of PH does not improve glucose tolerance. Mild worsening of glucose tolerance after treatment could be explained by an increase of the body mass index. These data, in accordance with our previous study, do not support the idea that PH is a secondary form of diabetes mellitus. PMID- 12899665 TI - Candidate genetic markers and the risk of restenosis after coronary angioplasty. AB - The aim of the present study was to test for possible associations between candidate gene polymorphisms and the risk of restenosis and recurrent restenosis after percutaneous transluminal coronary angioplasty (PTCA) without stenting. We followed up 511 PTCA patients, and restenosis and recurrent restenosis were defined according to angiographical criteria. Genotyping of the beta-fibrinogen 455 G/A, glycoprotein (GP) IIIa PlA1/PlA2, plasminogen activator inhibitor-1 (PAI 1) 4G/5G, factor V Leiden 1691 G/A, tumour necrosis factor alpha (TNFalpha) -238 G/A, TNFalpha -308 G/A, interleukin (IL)-1alpha -889 C/T, IL-1beta -511 C/T, methylenetetrahydrofolate reductase (MTHFR) 677 C/T and endothelial nitric oxide synthase (eNOS) 4 b/a gene polymorphisms was performed by PCR and restriction fragment-length-polymorphism-based techniques. One hundred and sixty patients (31.3%) developed restenosis and in 130 of these patients, of whom 123 were available for analysis, a second PTCA without stenting was performed. Of these patients, 35 (28.5%) developed recurrent restenosis. None of the investigated genotypes were associated with the risk of restenosis or recurrent restenosis after PTCA. The degree of stenosis before and immediately after PTCA and the severity of the lesion were independent predictors for restenosis after PTCA. In conclusion, there was no association between the beta-fibrinogen -455 G/A, GP IIIa PlA1/A2, PAI-1 4G/5G, factor V Leiden 1691 G/A, TNFalpha -238 G/A, TNFalpha 308 G/A, IL-1alpha -889 C/T, the IL-1beta -511 C/T, MTHFR 677 C/T and eNOS 4 b/a gene polymorphisms and the risk of restenosis after PTCA as well as recurrent restenosis after repeated PTCA. PMID- 12899666 TI - ECL cell histamine mobilization studied by gastric submucosal microdialysis in awake rats: methodological considerations. AB - The ECL cells are endocrine/paracrine cells in the acid-producing part of the stomach. They secrete histamine in response to circulating gastrin. Gastric submucosal microdialysis has been used to study ECL-cell histamine mobilization in awake rats. In the present study we assess the usefulness and limitations of the technique. Microdialysis probes were implanted in the gastric submucosa. Histological analysis of the stomach wall around the probe revealed a moderate, local inflammatory reaction 1-2 days after implantation; the inflammation persisted for at least 10 days. Experiments were conducted 3 days after the implantation. The "true" submucosal histamine concentration was determined by perfusing at different rates (the zero flow method) or with different concentrations of histamine at a constant rate (the no-net-flux method): in fasted rats it was calculated to be 87+/-5 (means+/-S.E.M.) nmol/l and 76+/-9 nmol/l, respectively. The corresponding histamine concentrations in fed rats were 93+/-5 and 102+/-8 nmol/l, respectively. With a perfusion rate of 74 microl/hr the recovery of submucosal histamine was 49%, at 34 microl/hr the recovery increased to 83%. At a perfusion rate below 20 microl/hr the microdialysate histamine concentration was close to the actual concentration in the submucosa. The ECL-cell histamine mobilization was independent of the concentrations of Ca2+ in the perfusion medium (0-3.4 mmol/l Ca2+). In one experiment, histamine mobilization in response to gastrin (10 nmol/kg/hr subcutaneously) was monitored in rats pretreated with prednisolone (60 mg/kg) or indomethacin (15 mg/kg). The two antiinflammatory agents failed to affect the concentration of histamine in the microdialysate either before or during the gastrin challenge, which was in accord with the observation that the inflammatory reaction was modest and that inflammatory cells were relatively few around the probe and in the wall of the probe. In another experiment, rats were given aminoguanidine (10 mg/kg) or metoprine (10 mg/kg) 4 hr before the start of gastrin infusion (5 nmol/kg/hr intravenously). Metoprine (inhibitor of histamine N-methyl transferase) did not affect the microdialysate histamine concentration, while aminoguanidine (inhibitor of diamine oxidase) raised both basal and gastrin-stimulated histamine concentrations. We conclude that microdialysis can be used to monitor changes in the concentration of histamine in the submucosa of the stomach, and that the inflammatory reaction to the probe is moderate and does not affect the submucosal histamine mobilization. PMID- 12899667 TI - The CB1 cannabinoid receptor agonist, HU-210, reduces levodopa-induced rotations in 6-hydroxydopamine-lesioned rats. AB - Parkinson's disease is a chronic neurodegenerative disease of the extrapyramidal system associated with dopaminergic neuronal loss in the basal ganglia. However, several other neurotransmitters, such as serotonin, gamma-amino-butyric acid and glutamate, are also related to the symptoms of Parkinson's disease patients and their response to levodopa treatment. The co-expression of cannabinoid and dopamine receptors in the basal ganglia suggests a potential role for endocannabinoids in the control of voluntary movement in Parkinson's disease. In the present study we treated unilaterally 2,4,5-trihydroxyphenethylamine (6 hydroxydopamine)-lesioned rats with the enantiomers of the synthetic cannabinoid 7-hydroxy-delta6-tetrahydrocannabinol 1,1-dimethylheptyl. Treatment with its (-)- (3R, 4R) enantiomer (code-name HU-210), a potent cannabinoid receptor type 1 agonist, reduced the rotations induced by levodopa/carbidopa or apomorphine by 34% and 44%, respectively. In contrast, treatment with the (+)- (3S, 4S) enantiomer (code-name HU-211), an N-methyl-D-aspartate antagonist, as well as the psychotropically inactive cannabis constituent: cannabidiol and its primary metabolite, 7-hydroxy-cannabinol, did not show any reduction of rotational behavior. Our results indicate that activation of the CB1 stimulates the dopaminergic system ipsilaterally to the lesion, and may have implications in the treatment of Parkinson's disease. PMID- 12899668 TI - Novel effect of N-palmitoyl-L-serine phosphoric acid on cytosolic Ca2+ levels in human osteoblasts. AB - The effect of N-palmitoyl-L-serine phosphoric acid (L-NASPA), which has been used as an inhibitor of lysophosphatidic acid receptors, on intracellular Ca2+ concentration ([Ca2+]i) in human osteosarcoma MG63 cells was measured by using fura-2. L-NASPA (0.1-10 microM) caused a rapid and transient plateau [Ca2+]i rise in a concentration-dependent manner (EC50=0.5 microM). The L-NASPA-induced [Ca2+]i rise was partly reduced by removal of extracellular Ca2+ but was not altered by L-type voltage-gated Ca2+ channel blockers. In Ca2+-free medium, thapsigargin, an inhibitor of the endoplasmic reticulum Ca2+-ATPase, induced a [Ca2+]i rise, after which the increasing effect of L-NASPA on [Ca2+]i was completely inhibited; also, pretreatment with L-NASPA partly reduced thapsigargin induced [Ca2+]i rise. U73122, an inhibitor of phospholipase C, abolished histamine (but not L-NASPA)-induced [Ca2+]i rise. Overnight incubation with 1 microM L-NASPA did not affect cell proliferation, but 10-20 microM L-NASPA exerted 4% and 15% inhibition, respectively. Collectively, L-NASPA rapidly increased [Ca2+]i in MG63 cells by evoking both extracellular Ca2+ influx and intracellular Ca2+ release, and is cytotoxic at higher concentrations. PMID- 12899669 TI - A screening study on the liability of eight different female sex steroids to inhibit CYP2C9, 2C19 and 3A4 activities in human liver microsomes. AB - The aim of this study was to screen the inhibitory potential of different clinically used oestrogen and progestin hormones on CYP2C9, 2C19 and 3A4 activities in human liver microsomes. The degree of inhibition by desogestrel, 3 ketodesogestrel, 17-beta-oestradiol, gestodene, aethinyloestradiol, medroxyprogesterone acetate, norethisterone or L-norgestrel were studied at 100 microM on losartan oxidation (CYP2C9), R-omeprazole 5'-hydroxylation (CYP2C19) and R-omeprazole sulphoxidation (CYP3A4) with a 10-min preincubation with NADPH in human liver microsomes prepared from 6 individual genotyped donor livers. Aethinyloestradiol was found to be a potent inhibitor (55% mean inhibition; 95% CI 32% to 79%) of losartan oxidation (CYP2C9) and R-omeprazole 5-hydroxylation (70%; 63% to 77%) (CYP2C19), while it had little effect on R-omeprazole sulphoxidation (CYP3A4) activity. 17-beta-Oestradiol did not produce significant inhibition on any of the studied enzyme activities. Of the progestin hormones studied, gestodene and 3-ketodesogestrel were potent inhibitors of CYP2C19 (57%; 47% to 67% and 51%; 29% to 45%) and CYP3A4 (45%; 30% to 59% and 40%; 19% to 62%), but had little effect on the CYP2C9 activity. In addition, medroxyprogesterone acetate was found to inhibit CYP2C9 (55%; 45% to 65%), while not having significant effect on 2C19 or 3A4. In conclusion, the liability of clinically used female sex steroids to inhibit CYP2C9, 2C19 and 3A4 activities in human liver microsomes is very distinctive and these differences among both the oestrogen and progestin hormones may, at least in part, explain the variable results from clinical trials examining inhibitory effects of hormone replacement therapy and oral contraceptives on drug metabolism. PMID- 12899670 TI - Gastrointestinal and systemic uptake of bismuth in mice after oral exposure. AB - Bismuth compounds have been used in medicine for more than 200 years. In recent years, bismuth has gained renewed interest as a remedy for eradication of gastrointestinal pathogens, especially Helicobacter pylori. In this study we describe the anatomical distribution of bismuth in the gastrointestinal tract and other organs after oral exposure in a mouse model. After exposure of the experimental animals to ranitidine bismuth citrate or bismuth citrate, we used the autometallographic silver enhancement technique to demonstrate the presence of bismuth in tissue samples from the gastrointestinal tract, liver, spleen, thymus, kidney and lymph nodes. We exposed cultured murine peritoneal macrophages to bismuth citrate and examined the bismuth accumulation over time. We found that in the mouse bismuth is absorbed systemically after a single dose of either compound, ranitidine bismuth more easily than bismuth citrate. Uptake could be shown in the stomach, duodenum, ileum and kidney for hours after exposure. Weeks after the exposure, deposits of bismuth were found in lymph nodes, liver, spleen and kidney as well as in macrophages in the gastrointestinal lamina propria. At the subcellular level, bismuth was found exclusively in lysosomes, primarily in macrophages and dendritic cells. Subsequent analyses of macrophage cultures showed lysosomal accumulations to be time and dose dependent. PMID- 12899671 TI - Modulation of GABAA receptors and neuropeptide secretion by the neurosteroid allopregnanolone in posterior and intermediate pituitary. AB - A number of neurosteroids bind to GABAA receptors and alter their responsiveness to neurotransmitters. Considerable effort has been devoted to understanding how this form of receptor modulation alters inhibitory synaptic function. Neurosteroid-sensitive GABAA receptors have also been demonstrated in many endocrine cells, but little is known about how neurosteroids modulate the release of hormones. Here, the action of allopregnanolone, a neurosteroid that enhances GABAA receptor-mediated responses, was investigated in posterior pituitary nerve terminals and intermediate pituitary endocrine cells. Patch clamp recordings showed that GABA-evoked currents were enhanced to similar degrees and with similar concentration dependences in both locations. An organ bath preparation of the neurointermediate lobe was used to investigate drug effects on secretion of vasopressin and alpha-melanocyte stimulating hormone. GABA increased the basal release of vasopressin and alpha-melanocyte stimulating hormone from the posterior and intermediate pituitary lobe, respectively, an effect that could be blocked by picrotoxinin. Vasopressin release evoked by electrical stimulation was also examined, and a small statistically significant inhibition by 5 microM GABA was observed. Allopregnanolone increased the basal release of vasopressin, and this effect was blocked by the GABAA receptor antagonist picrotoxinin. Allopregnanolone had no effect in conjunction with GABA. In contrast to the posterior lobe, allopregnanolone had no effect on release from the intermediate lobe. Thus, allopregnanolone in physiological relevant concentrations modulates GABAA receptors in both the posterior and intermediate lobes, but only affects hormone release in the posterior lobe. PMID- 12899672 TI - Survival following a metformin overdose of 63 g: a case report. PMID- 12899673 TI - Chronic ethanol administration and withdrawal decreases 5-HT1A mRNA, but not 5 HT4 expression in the rat hippocampus. PMID- 12899675 TI - Palatable solutions during paradoxical sleep deprivation: reduction of hypothalamic-pituitary-adrenal axis activity and lack of effect on energy imbalance. AB - Paradoxical sleep deprivation (PSD) induces increased energy expenditure in rats, insofar as rats eat more but loose weight throughout the deprivation period. In the present study, rats were offered water, saccharin or sucrose to drink during the deprivation period, since it has been proposed that carbohydrates reduce the hypothalamic-pituitary-adrenal (HPA) axis response to stress. Rats were submitted to the flower pot technique for 96 h. During the PSD period, they were weighed daily and food and fluid intake was assessed twice a day. At the end of the PSD period, rats were killed and plasma concentrations of glucose, adrenocorticotropic hormone (ACTH) and corticosterone were assayed. Compared to their control counterparts, all paradoxical sleep-deprived rats consumed more food, but lost weight. Paradoxical sleep-deprived rats given sucrose drank more than their control counterparts (especially in the light phase of the light/dark cycle). Paradoxical sleep-deprived rats showed increased food intake during all periods throughout the experiment, with peak intake during the dark phase and nadir during the light phase of the light/dark cycle. All paradoxical sleep deprived rats showed lower glucose plasma levels than control rats and increased relative adrenal weight. However, when given saccharin or sucrose, paradoxical sleep-deprived rats showed lower concentrations of ACTH and corticosterone than their water-provided counterparts, indicating that palatable fluids were capable of lowering HPA axis activation produced by PSD. The fact that PSD induced energy imbalance regardless of the relative attenuation of the HPA axis activity produced by saccharin or sucrose suggests that the HPA axis may play only a secondary role in this phenomenon, and that other mechanisms may account for this effect. The data also suggest that supply of palatable fluids can be an additional modification to reduce the stress of the flower pot method. PMID- 12899676 TI - Leptin-target neurones of the rat hypothalamus express somatostatin receptors. AB - Hypothalamic leptinoceptive neurones can be visualized by histochemical demonstration of leptin-induced nuclear translocation of the signalling molecule STAT3. We investigated the relationship of the leptinoceptive neurones to the somatostatin signalling system. With double-labelling immunohistochemistry, we studied the colocalization of leptin-activated transcription factor, STAT3, with somatostatin receptor subtypes, sst1, sst2A, sst2B, sst3 and sst4, or the neuropeptide itself, in the rat hypothalamus. Immunoreactivity for all the entities was widely distributed throughout the entire hypothalamus. Despite the wide distribution, only few cases of colocalization of somatostatin with leptin activated STAT3 were detected in the paraventricular, arcuate and dorsomedial nuclei. A moderate to high degree of colocalization of nuclear STAT3 and all investigated subtypes of somatostatin receptors was found in the lateral and dorsal hypothalamic areas and in the dorsomedial hypothalamic nucleus. Immunoreactivity for sst1, sst2B and sst4 was present in STAT3-containing nuclei of the paraventricular, periventricular, arcuate and ventromedial hypothalamic neurones, as well as in the retrochiasmatic and posterior hypothalamic areas. Despite the wide distribution of sst2A in the rat hypothalamus, few events of colocalization with leptin-activated STAT3 were observed in the dorsomedial nucleus and in the lateral and dorsal hypothalamic areas only. Many leptin responsive neurones of the dorsal, lateral, periarcuate, perifornical and posterior hypothalamic areas, as well as in the ventromedial and dorsomedial hypothalamic nuclei, displayed sst3 immunoreactivity at their neuronal cilia. These results provide strong anatomical evidence for the direct interaction of leptin and the somatostatin systems in neuroendocrine control loops such as the energy homeostasis, growth or stress response. PMID- 12899677 TI - Evidence that gonadotropin-releasing hormone II is not a physiological regulator of gonadotropin secretion in mammals. AB - Gonadotropin-releasing hormone (GnRH)-II stimulates luteinizing hormone (LH) and follicle-stimulating hormone (FSH) secretion when administered at high doses in mammals, and this effect has been assumed to be mediated through the GnRH-II receptor expressed on gonadotropes. This study used two selective GnRH-I receptor antagonists to test the alternative hypothesis that GnRH-II acts through the GnRH I receptor to elicit gonadotropin secretion. The antagonist, antide, was used to characterize the receptor-relay because it was a pure antagonist in vitro based on inositol phosphate responses in COS-7 cells transfected with either mammalian GnRH-I and GnRH-II receptors and, in vivo, potently antagonized the gonadotropin releasing effect of a single injection of 250 ng GnRH-I in our sexually inactive sheep model. In a series of studies in sheep, antide (i). blocked the acute LH response to a single injection of GnRH-II (20 microg antide: 10 microg GnRH-II); (ii). blocked both the acute, pulsatile LH response and the FSH priming response to 2-hourly injections of GnRH-II over 36 h (100 microg antide/8 h: 4 microg GnRH II/2 h); and (iii). chronically blocked both the pulsatile LH response and the marked FSH priming response to 4-hourly injections of GnRH-II over 10 days (75 microg antide/8 h: 4 microg GnRH-II/4 h). In two final experiments, the GnRH-I antagonist 135-18, shown previously to agonize the mammalian GnRH-II receptor, blocked the gonadotropin-releasing effects of GnRH-I (250 ng) but failed to elicit an LH response when given alone, and simultaneous administration of GnRH II (250 ng) failed to alter the LH-releasing effect of GnRH-I (50-500 ng). These data thus support our hypothesis. Based on additional literature, it is unlikely that the GnRH-II decapeptide is a native regulator of the gonadotrope in mammals. PMID- 12899678 TI - Receptor activity modifying protein 2 distribution in the rat central nervous system and regulation by changes in blood pressure. AB - The neuropeptide, adrenomedullin, acts in the central nervous system (CNS) to regulate blood pressure, at least partly through an adrenomedullin receptor which is composed of receptor activity modifying protein 2 (RAMP-2) and calcitonin receptor-like receptor (CRLR). We used in situ hybridization to localize RAMP-2 mRNA throughout the brain, and we performed reverse transcription-polymerase chain reaction to detect CRLR mRNA in the brain. We found that RAMP-2 mRNA is expressed in numerous areas, including autonomic nuclei such as the paraventricular, supraoptic, arcuate and ventromedial nuclei, as well as the nucleus of the solitary tract (NTS), area postrema and dorsal motor nucleus of the vagus. Many regions expressing RAMP-2 mRNA also express low levels of CRLR mRNA. We examined changes in the mRNA expression of RAMP-2 and preproadrenomedullin in the brain in response to blood pressure manipulations. Rats received intravenous infusions of nitroprusside or phenylephrine to decrease or increase blood pressure, respectively. Decreased blood pressure elicited an increase in RAMP-2 mRNA expression in the NTS and a decrease in preproadrenomedullin mRNA expression in the paraventricular nucleus (PVN). Increased blood pressure elicited a decrease in RAMP-2 mRNA expression in the PVN and NTS. The CNS distribution and modulation of adrenomedullin signalling components by changes in blood pressure provide anatomical and physiological evidence for a homeostatic role for adrenomedullin in the brain. PMID- 12899679 TI - Sleep loss reduces diurnal rhythm amplitude of leptin in healthy men. AB - The aim of the current study was to investigate the effects of sleep loss on the diurnal rhythm of circulating leptin levels. An indwelling forearm catheter was used to sample blood at 90-min intervals for a total of 120 h, which included 88 h of sustained sleeplessness, in 10 healthy men. The diurnal amplitude of leptin was reduced during total sleep deprivation and returned toward normal during the period of recovery sleep. This finding provides evidence that sleep influences the nocturnal leptin profile, and may have implications for the understanding of the role of sleep in metabolic regulation and the aetiologies of obesity and the night eating syndrome. PMID- 12899680 TI - Immunocytochemical detection of progesterone receptor in the female rabbit forebrain: distribution and regulation by oestradiol and progesterone. AB - There is no information on the neuroanatomical distribution of the progesterone receptor (PR) in the rabbit. Therefore, we mapped the distribution of PR immunoreactive cells in the forebrain of ovariectomized female rabbits. Vehicle injected ovariectomized rabbits showed PR-immunoreactive cells only in the infundibular nucleus (IN) and nucleus X (lateral to the ventromedial hypothalamic nucleus). The injection of oestradiol benzoate (EB; 5 micro g/day for 5 days) increased the number of PR-immunoreactive cells in the IN and in three nuclei of the preoptic region (periventricular, medial, and principal). Abundant PR were also found in the paraventricular nucleus and nucleus X. Administration of progesterone (10 mg/day) for 3 days to EB-treated rabbits (a treatment that induces digging behaviour for the maternal nest and suppresses sexual receptivity and scent-marking) eliminated PR-immunoreactivity from all brain areas analysed except the IN. Thus, one-third of the number of cells seen in the ovariectomized + EB condition persisted in this region despite progesterone injections. Withdrawal of progesterone (and continuation of EB) for 5 (but not for 2) days (in a schedule similar to the one that induces straw-carrying and hair-pulling for the maternal nest) increased the number of PR-immunoreactive cells in all regions analysed. These results show that restricted regions of the female rabbit forebrain express abundant PR which are either: (i). up-regulated by oestradiol and down-regulated by progesterone; (ii). oestradiol-insensitive and down regulated by progesterone; or (iii). insensitive to both oestradiol and progesterone. PMID- 12899681 TI - Early expression of chicken gonadotropin-releasing hormone-1 in the developing chick. AB - Gonadotropin-releasing hormone (GnRH), which is essential for reproductive function, is made by neurones that migrate from the nasal region into the brain during early embryonic development. This migration begins in chick when the olfactory pit is formed. This is approximately the time that GnRH neurones can be detected immunocytochemically. The present study investigated (i). how early in development the GnRH gene is expressed and (ii). the sites of its expression. Accordingly, reverse transcriptase-polymerase chain reaction (PCR) and in situ hybridization were performed on chick embryos before gastrulation up until the stage by which GnRH neurones have begun to migrate into the central nervous system. Primers were made to the 5'- and 3'-UTR region of the message for cGnRH I, the form of the peptide that is essential for reproductive function in the chicken. PCR product was found in all stages and the sequences of products from all stages were identical. Thus, the GnRH gene is expressed continuously throughout embryonic development. In situ hybridization with a digoxygenin labelled riboprobe revealed staining along the primitive streak immediately before gastrulation. In later stages, cGnRH-I gene expression was seen in association with the anterior neural ridge. The expression was subsequently restricted to a narrow, clearly defined region, which is associated with the presumptive nasal cavity and olfactory placode. Later, GnRH neurones could be seen in their migratory routes by both in situ hybridization and immunocytochemistry. Expression of the GnRH gene has been described in preimplantation stages in mammals and there is evidence that the neuropeptide plays a role in formation and maintenance of the placenta. What role (if any) it may play in early avian development remains unknown. The demonstration of sites of GnRH expression during the early period of neurulation suggests that GnRH neurones arise before olfactory placode formation. PMID- 12899682 TI - Impaired somatodendritic responses to pituitary adenylate cyclase-activating polypeptide (PACAP) of supraoptic neurones in PACAP type I -receptor deficient mice. AB - The role of pituitary adenylate cyclase-activating polypeptide (PACAP) type I receptor (PAC1 receptor) in regulating hypothalamic supraoptic neurones was investigated using PAC1 receptor-deficient male mice (PAC1-/-). The effects of PACAP on [Ca2+]i were investigated in freshly dissociated supraoptic neurones and on the somatodendritic release of vasopressin and oxytocin, examined on intact supraoptic nuclei. In supraoptic neurones from wild-type mice (PAC1+/+), 100 nm PACAP induced an increase in [Ca2+]i and release of vasopressin and oxytocin, whereas in heterozygous (PAC1+/-) and null-mutant mice (PAC1-/-), PACAP was much less effective. PACAP had no effect on these two parameters when applied to isolated neurohypophysial nerve terminals of PAC1+/+ and PAC1-/- mice, and rats. In conclusion, the PAC1 receptor is solely responsible for the PACAP-induced [Ca2+]i signalling and secretion of vasopressin and oxytocin in the somatodendritic region of supraoptic neurones. PMID- 12899683 TI - Androgen 5-alpha-reductase type 2 is highly expressed and active in rat spinal cord motor neurones. AB - Spinal cord motoneurones express high levels of androgen receptor. However, in responsive tissue, the effects of testosterone is often mediated by the more potent androgenic derivative 5-alpha-dihydrotestosterone (DHT). This compound is formed in androgen target cells by the enzyme 5-alpha-reductase. Two isoforms of the 5-alpha-reductase, with limited degree of homology, have been cloned, type 1 and type 2. The low affinity-constitutive type 1 isoenzyme is widely distributed in the body; the high affinity-androgen regulated 5-alpha-reductase type 2 is confined to androgen-dependent structures and shows a peculiar pH optimum at acidic values. We have previously shown that high levels of 5-alpha-reductase activity are detectable in rat spinal cord. Here, using reverse transcriptase polymerase chain reaction, we show that both isoforms are expressed in the whole spinal cord of the rat. The enzymatic pH optimum measured in immortalized spinal cord motoneurones (NSC34) is typical of the type 2 isoenzyme. Using in situ hybridization technique, we found that 5-alpha-reductase type 2 is confined to the motoneuronal cells of the anterior horns of the rat spinal cord, the cells that also are known to express high levels of androgen receptor. Because of the close association of androgen receptor and 5-alpha alpha-reductase type 2, motoneuronal cells should be considered as target cells for androgens. PMID- 12899684 TI - mu-Opioid receptor modulates peptide release from rat neurohypophysial terminals by inhibiting Ca(2+) influx. AB - The activation of opioid receptors in neurones of the central nervous system leads to a variety of effects including the modulation of diuresis and parturition, processes that are directly controlled by the hypothalamic neurohypophysial system (HNS). The effects of mu-opioid receptor activation on peptide release, voltage-gated Ca2+ currents and intracellular calcium levels ([Ca2+]i) were studied in isolated nerve terminals of the HNS. The mu-receptor agonist, DAMGO ([d-Ala2,N-Me-Phe4,Gly5-ol]-enkephalin) inhibited high K+-induced peptide release in a dose-dependent manner, with oxytocin release being more sensitive to block than vasopressin release at all concentrations tested. The addition of the mu-receptor antagonist CTOP (d-Phe-Cys-Tyr-d-Trp-Orn-Thr-Pen-Thr amide) was able to overcome the inhibitory effects of DAMGO. By contrast to previous results, voltage-gated Ca2+ currents were sensitive to blockage by DAMGO and this inhibition was also prevented by CTOP. Furthermore, [Ca2+]i measurements with Fura-2 corroborated the inhibition by DAMGO of calcium entry and its reversal by the micro -receptor antagonist in these nerve terminals. Thus, the decrease in neuropeptide release, particularly for oxytocin, induced by the activation of mu-opioid receptors in neurohypophysial terminals is mediated, at least in part, by a corresponding decrease in Ca2+ entry due to the inhibition of voltage-gated Ca2+ channels. PMID- 12899685 TI - Responses of cells in the rat suprachiasmatic nucleus in vivo to stimulation of afferent pathways are different at different times of the light/dark cycle. AB - Conventional extracellular recordings were made from single cells in the suprachiasmatic nucleus (SCN) region of the anaesthetized rat. Each cell was tested for its response to stimulation at three sites; the contralateral optic nerve, the ipsilateral supraoptic nucleus (SON) or the ipsilateral arcuate nucleus (ARC) to determine whether the behaviour of the synapses in the SCN was different at different times. Responses to stimulation were tested once each hour and assessed by creating peristimulus time histograms. Excitatory, inhibitory or complex (consisting of more than one component) responses were seen. The responses of some cells that were recorded for several hours changed with time. Changes were seen in the responses of SCN cells to stimulation of the ARC (31/91 cells) and the SON (26/90 cells) regions, but only rarely to stimulation of the optic nerve (2/72 cells). Such differences in proportion are unlikely to have occurred by chance (P < 0.001; chi-square test). Changes seen included the appearance of both excitatory and inhibitory responses in cells that were initially unresponsive. In some cells, one component of a complex response remained constant while another component changed with time. When the cells in the SCN were treated as a group, the proportion of excitatory, inhibitory or complex responses to ARC stimulation did not remain constant throughout the light/dark cycle (P = 0.014; chi-square test). The proportion of excitatory, inhibitory or complex responses to SON and optic nerve stimulation showed no significant variation with the light/dark cycle. If a change in response can be interpreted as a change in the behaviour of a neural connection, the results imply that some of the projections to the SCN from within the hypothalamus change at different times of the light/dark cycle, whereas no change could be seen in the input from the optic nerve. Thus, some of the connections of the SCN appear not to be hard wired, but change rapidly with time. PMID- 12899686 TI - Abstracts of the 19th Congress of the International Association of Paediatric Dentistry. New Orleans, Louisiana, USA. 15-18 October 2003. PMID- 12899687 TI - Fidelity and spatio-temporal control in MAP kinase (ERKs) signalling. AB - The mitogen activated protein (MAP) kinase module: (Raf -->MEK-->ERKs) is central to the control of cell growth, cell differentiation and cell survival. The fidelity of signalling and the spatio-temporal activation are key determinants in generating precise biological responses. The fidelity is ensured by scaffold proteins - protein kinase 'insulators' - and by specific docking sites. The duration and the intensity of the response are in part controlled by the compartmentalization of the signalling molecules. Growth factors promote rapid nuclear translocation and persistent activation of p42/p44 MAP kinases, respectively and ERK2/ERK1, during the entire G1 period with an extinction during the S-phase. These features are exquisitely controlled by the temporal induction of the MAP kinase phosphatases, MKP1-3. MKP1 and 2 induction is strictly controlled by the activation of the MAP kinase module providing evidence for an auto-regulatory mechanism. This negative regulatory loop is further enhanced by the capacity of p42/p44 MAPK to phosphorylate MKP1 and 2. This action reduces the degradation rate of MKPs through the ubiquitin-proteasomal system. Whereas the two upstream kinases of the module (Raf and MEK) remain cytoplasmic, ERKs (anchored to MEK in the cytoplasm of resting cells) rapidly translocate to the nucleus upon mitogenic stimulation. This latter process is rapid, reversible and controlled by the strict activation of the MAPK cascade. Following long-term MAPK stimulation, p42/p44 MAPKs progressively accumulate in the nucleus in an inactive form. Therefore we propose that the nucleus represents a site for ERK action, sequestration and signal termination. With the generation of knockdown mice for each of the ERK isoforms, we will illustrate that besides controlling cell proliferation the ERK cascade also controls cell differentiation and cell behaviour. PMID- 12899688 TI - Tritium planigraphy comparative structural study of tobacco mosaic virus and its mutant with altered host specificity. AB - Spatial organization of wild-type (strain U1) tobacco mosaic virus (TMV) and of the temperature-sensitive TMV ts21-66 mutant was compared by tritium planigraphy. The ts21-66 mutant contains two substitutions in the coat protein (Ile21-->Thr and Asp66-->Gly) and, in contrast with U1, induces a hypersensitive response (formation of necroses) on the leaves of plants bearing a host resistance gene N' (for example Nicotiana sylvestris); TMV U1 induces systemic infection (mosaic) on the leaves of such plants. Tritium distribution along the coat protein (CP) polypeptide chain was determined after labelling of both isolated CP preparations and intact virions. In the case of the isolated low-order (3-4S) CP aggregates no reliable differences in tritium distribution between U1 and ts21-66 were found. But in labelling of the intact virions a significant difference between the wild type and mutant CPs was observed: the N-terminal region of ts21-66 CP incorporated half the amount of tritium than the corresponding region of U1 CP. This means that in U1 virions the CP N-terminal segment is more exposed on the virion surface than in ts21-66 virions. The possibility of direct participation of the N-terminal tail of U1 CP subunits in the process of the N' hypersensitive response suppression is discussed. PMID- 12899689 TI - Diversity, taxonomy and evolution of medium-chain dehydrogenase/reductase superfamily. AB - A comprehensive, structural and functional, in silico analysis of the medium chain dehydrogenase/reductase (MDR) superfamily, including 583 proteins, was carried out by use of extensive database mining and the blastp program in an iterative manner to identify all known members of the superfamily. Based on phylogenetic, sequence, and functional similarities, the protein members of the MDR superfamily were classified into three different taxonomic categories: (a) subfamilies, consisting of a closed group containing a set of ideally orthologous proteins that perform the same function; (b) families, each comprising a cluster of monophyletic subfamilies that possess significant sequence identity among them and might share or not common substrates or mechanisms of reaction; and (c) macrofamilies, each comprising a cluster of monophyletic protein families with protein members from the three domains of life, which includes at least one subfamily member that displays activity related to a very ancient metabolic pathway. In this context, a superfamily is a group of homologous protein families (and/or macrofamilies) with monophyletic origin that shares at least a barely detectable sequence similarity, but showing the same 3D fold. The MDR superfamily encloses three macrofamilies, with eight families and 49 subfamilies. These subfamilies exhibit great functional diversity including noncatalytic members with different subcellular, phylogenetic, and species distributions. This results from constant enzymogenesis and proteinogenesis within each kingdom, and highlights the huge plasticity that MDR superfamily members possess. Thus, through evolution a great number of taxa-specific new functions were acquired by MDRs. The generation of new functions fulfilled by proteins, can be considered as the essence of protein evolution. The mechanisms of protein evolution inside MDR are not constrained to conserve substrate specificity and/or chemistry of catalysis. In consequence, MDR functional diversity is more complex than sequence diversity. MDR is a very ancient protein superfamily that existed in the last universal common ancestor. It had at least two (and probably three) different ancestral activities related to formaldehyde metabolism and alcoholic fermentation. Eukaryotic members of this superfamily are more related to bacterial than to archaeal members; horizontal gene transfer among the domains of life appears to be a rare event in modern organisms. PMID- 12899690 TI - Characterization of the serotoninergic system in the C57BL/6 mouse skin. AB - We showed expression of the tryptophan hydroxylase gene and of tryptophan hydroxylase protein immunoreactivity in mouse skin and skin cells. Extracts from skin and melanocyte samples acetylated serotonin to N-acetylserotonin and tryptamine to N-acetyltryptamine. A different enzyme from arylalkylamine N acetyltransferase mediated this reaction, as this gene was defective in the C57BL6 mouse, coding predominantly for a protein without enzymatic activity. Serotonin (but not tryptamine) acetylation varied according to hair cycle phase and anatomic location. Serotonin was also metabolized to 5-hydroxytryptophol and 5-hydroxyindole acetic acid, probably through stepwise transformation catalyzed by monoamine oxidase, aldehyde dehydrogenase and aldehyde reductase. Activity of the melatonin-forming enzyme hydroxyindole-O-methyltransferase was notably below detectable levels in all samples of mouse corporal skin, although it was detectable at low levels in the ears and in Cloudman melanoma (derived from the DBA/2 J mouse strain). In conclusion, mouse skin has the molecular and biochemical apparatus necessary to produce and metabolize serotonin and N acetylserotonin, and its activity is determined by topography, physiological status of the skin, cell type and mouse strain. PMID- 12899691 TI - An Escherichia coli twin-arginine signal peptide switches between helical and unstructured conformations depending on the hydrophobicity of the environment. AB - The Tat system catalyzes the transport of folded globular proteins across the bacterial plasma membrane and the chloroplast thylakoid. It recognizes cleavable signal peptides containing a critical twin-arginine motif but little is known of the overall structure of these peptides. In this report, we have analyzed the secondary structure of the SufI signal peptide, together with those of two nonfunctional variants in which the region around the twin-arginine, RRQFI, is replaced by KKQFI or RRQAA. Circular dichroism studies show that the SufI peptide exists as an unstructured peptide in aqueous solvent with essentially no stable secondary structure. In membrane-mimetic environments such as SDS micelles or water/trifluoroethanol, however, the peptide adopts a structure containing up to about 40% alpha-helical content. Secondary structure predictions and molecular modelling programs strongly suggest that the helical region begins at, or close to, the twin-arginine motif. Studies on the thermal stability of the helix demonstrate a sharp transition between the unstructured and helical states, suggesting that the peptide exists in one of two distinct states. The two nonfunctional peptides exhibit almost identical spectra and properties to the wild-type SufI peptide, indicating that it is the arginine sidechains, and not their contribution to the helical structure, that are critical in this class of peptide. PMID- 12899692 TI - Multiple enzymic activities of human milk lactoferrin. AB - Lactoferrin (LF) is a Fe3+-binding glycoprotein, first recognized in milk and then in other human epithelial secretions and barrier fluids. Many different functions have been attributed to LF, including protection from iron-induced lipid peroxidation, immunomodulation and cell growth regulation, DNA binding, and transcriptional activation. Its physiological role is still unclear, but it has been suggested to be responsible for primary defense against microbial and viral infection. We present evidence that different subfractions of purified human milk LF possess five different enzyme activities: DNase, RNase, ATPase, phosphatase, and malto-oligosaccharide hydrolysis. LF is the predominant source of these activities in human milk. Some of its catalytically active subfractions are cytotoxic and induce apoptosis. The discovery that LF possesses these activities may help to elucidate its many physiological functions, including its protective role against microbial and viral infection. PMID- 12899693 TI - Human intrinsic factor expressed in the plant Arabidopsis thaliana. AB - Intrinsic factor (IF) is the gastric protein that promotes the intestinal uptake of vitamin B12. Gastric IF from animal sources is used in diagnostic tests and in vitamin pills. However, administration of animal IF to humans becomes disadvantageous because of possible pathogenic transmission and contamination by other B12 binders. We tested the use of recombinant plants for large-scale production of pathogen-free human recombinant IF. Human IF was successfully expressed in the recombinant plant Arabidopsis thaliana. Extract from fresh plants possessed high B12-binding capacity corresponding to 70 mg IF per 1 kg wet weight. The dried plants still retained 60% of the IF activity. The purified IF preparation consisted of a 50-kDa glycosylated protein with the N-terminal sequence of mature IF. Approximately one-third of the protein was cleaved at the internal site em leader PSNP downward arrow GPGP. The key properties of the preparation obtained were identical to those of native IF: the binding curves of vitamin B12 to recombinant IF and gastric IF were the same, as were those for a B12 analogue cobinamide, which binds to IF with low affinity. The absorbance spectra of the vitamin bound to recombinant IF and gastric IF were alike, as was the interaction of recombinant and native IF with the specific receptor cubilin. The data presented show that recombinant plants have a great potential as a large scale source of human IF for analytical and therapeutic purposes. PMID- 12899694 TI - Mapping the functional domain of the prion protein. AB - Prion diseases such as Creutzfeldt-Jakob disease are possibly caused by the conversion of a normal cellular glycoprotein, the prion protein (PrPc) into an abnormal isoform (PrPSc). The process that causes this conversion is unknown, but to understand it requires a detailed insight into the normal activity of PrPc. It has become accepted from results of numerous studies that PrPc is a Cu-binding protein and that its normal function requires Cu. Further work has suggested that PrPc is an antioxidant with an activity like that of a superoxide dismutase. We have shown in this investigation that this activity is optimal for the whole protein and that deletion of parts of the protein reduce or abolish this activity. The protein therefore contains an active domain requiring certain regions such as the Cu-binding octameric repeat region and the hydrophobic core. These regions show high evolutionary conservation fitting with the idea that they are important to the active domain of the protein. PMID- 12899695 TI - Glucose sensing in the intestinal epithelium. AB - Dietary sugars regulate expression of the intestinal Na+/glucose cotransporter, SGLT1, in many species. Using sheep intestine as a model, we showed that lumenal monosaccharides, both metabolisable and nonmetabolisable, regulate SGLT1 expression. This regulation occurs not only at the level of transcription, but also at the post-transcriptional level. Introduction of d-glucose and some d glucose analogues into ruminant sheep intestine resulted in > 50-fold enhancement of SGLT1 expression. We aimed to determine if transport of sugar into the enterocytes is required for SGLT1 induction, and delineate the signal transduction pathways involved. A membrane impermeable d-glucose analogue, di(glucos-6-yl)poly(ethylene glycol) 600, was synthesized and infused into the intestines of ruminant sheep. SGLT1 expression was determined using transport studies, Northern and Western blotting, and immunohistochemistry. An intestinal cell line, STC-1, was used to investigate the signalling pathways. Intestinal infusion with di(glucos-6-yl)poly(ethylene glycol) 600 led to induction of functional SGLT1, but the compound did not inhibit Na+/glucose transport into intestinal brush-border membrane vesicles. Studies using cells showed that increased medium glucose up-regulated SGLT1 abundance and SGLT1 promoter activity, and increased intracellular cAMP levels. Glucose-induced activation of the SGLT1 promoter was mimicked by the protein kinase A (PKA) agonist, 8Br-cAMP, and was inhibited by H-89, a PKA inhibitor. Pertussis toxin, a G-protein (Gi) specific inhibitor, enhanced SGLT1 protein abundance to levels observed in response to glucose or 8Br-cAMP. We conclude that lumenal glucose is sensed by a glucose sensor, distinct from SGLT1, residing on the external face of the lumenal membrane. The glucose sensor initiates a signalling pathway, involving a G protein-coupled receptor linked to a cAMP-PKA pathway resulting in enhancement of SGLT1 expression. PMID- 12899696 TI - Evidence for proteasome dysfunction in cytotoxicity mediated by anti-Ras intracellular antibodies. AB - Anti-Ras intracellular antibodies inhibit cell proliferation in vivo by sequestering the antigen and diverting it from its physiological location [Lener, M., Horn, I. R., Cardinale, A., Messina, S., Nielsen, U.B., Rybak, S.M., Hoogenboom, H.R., Cattaneo, A., Biocca, S. (2000) Eur. J. Biochem.267, 1196 1205]. Here we demonstrate that strongly aggregating single-chain antibody fragments (scFv), binding to Ras, induce apoptosis, and this effect is strictly related to the antibody-mediated aggregation of p21Ras. Proteasomes are quickly recruited to the newly formed aggregates, and their activity is strongly inhibited. This leads to the formation of aggresome-like structures, which become evident in the vast majority of apoptotic cells. A combination of anti-Ras scFv fragments with a nontoxic concentration of the proteasome inhibitor, lactacystin, markedly increases proteasome dysfunction and apoptosis. The dominant-negative H ras (N17-H-ras), which is mostly soluble and does not induce aggresome formation or inhibit proteasome activity, only affects cell viability slightly. Together, these observations suggest a mechanism linking antibody-mediated Ras aggregation, impairment of the ubiquitin-proteasome system, and cytotoxicity. PMID- 12899697 TI - Pherokine-2 and -3. AB - Drosophila is a powerful model system to study the regulatory and effector mechanisms of innate immunity. To identify molecules induced in the course of viral infection in this insect, we have developed a model based on intrathoracic injection of the picorna-like Drosophila C virus (DCV). We have used MALDI-TOF mass spectrometry to compare the hemolymph of DCV infected flies and control flies. By contrast with the strong humoral response triggered by injection of bacteria or fungal spores, we have identified only one molecule induced in the hemolymph of virus infected flies. This molecule, pherokine-2 (Phk-2), is related to OS-D/A10 (Phk-1), which was previously characterized as a putative odor/pheromone binding protein specifically expressed in antennae. The virus induced molecule is also similar to the product of the gene CG9358 (Phk-3), which is induced by septic injury. Both Phk-2 and Phk-3 are strongly expressed during metamorphosis, suggesting that they may participate in tissue-remodeling. PMID- 12899698 TI - Regulation of Cyr61/CCN1 gene expression through RhoA GTPase and p38MAPK signaling pathways. AB - Cysteine-rich protein 61 (Cyr61/CCN1) is an angiogenic factor and a member of a family of growth factor-inducible immediate-early genes with functions in cell adhesion, proliferation and differentiation. We investigated the regulatory mechanisms and signaling pathways involved in Cyr61/CCN1gene activation in smooth muscle cells. Treatment of these cells with sphingosine 1-phosphate (S1P), a bioactive lysolipid, increased rapidly but transiently the expression of the Cyr61/CCN1 gene at both the mRNA and protein levels. Cyr61/CCN1 mRNA stability was not altered but the transcription rate of the Cyr61/CCN1 gene was increased fivefold in isolated nuclei from S1P-stimulated cells indicating that the level of control is primarily transcriptional. Transfection experiments showed that a 936-bp promoter fragment of the human Cyr61/CCN1 gene is functional and induces a reporter gene activity in S1P-treated cells. Using a combination of cis-element mutagenesis and expression of dominant negative inhibitors of transcription factors, we showed that both a CRE and AP-1 site and their cognate transcription factors, cAMP response element binding protein (CREB) and AP-1, were responsible for the promoter activity in S1P-stimulated cells. Furthermore, by using either pharmacological inhibitors or active forms of known signaling molecules, we showed that inducible Cyr61/CCN1 gene expression occurs through RhoA GTPase and that additional signaling through the p38 pathway is required. In particular, p38 seems to regulate Cyr61/CCN1 promoter activity through modulation of phosphorylation of CREB and the CREB kinase, MSK1. These findings demonstrate the transcriptional regulation of the Cyr61/CCN1 gene and provide clues to the signaling molecules and transcription factors involved in such regulation. PMID- 12899699 TI - Purification and properties of a new S-adenosyl-L-methionine:flavonoid 4'-O methyltransferase from carnation (Dianthus caryophyllus L.). AB - A new enzyme, S-adenosyl-l-methionine:flavonoid 4'-O-methyltransferase (EC 2.1.1. ) (F 4'-OMT), has been purified 1 399-fold from the tissues of carnation (Dianthus caryophyllus L). The enzyme, with a molecular mass of 43-45 kDa and a pI of 4.15, specifically methylates the hydroxy substituent in 4'-position of the flavones, flavanones and isoflavones in the presence of S-adenosyl-l-methionine. A high affinity for the flavone kaempferol was observed (Km = 1.7 micro m; Vmax = 95.2 micro mol.min-1.mg-1), while other 4'-hydroxylated flavonoids proved likewise to be suitable substrates. Enzyme activity had no apparent Mg++ requirement but was inhibited by SH-group reagents. The optimum pH value for F 4' OMT activity was found to be around neutrality. Kinetic analysis of the enzyme bi substrate reaction indicates a Ping-Pong mechanism and excludes the formation of a ternary complex. The F 4'-OMT activity was increased, in both in vitro and in vivo carnation tissues, by the inoculation with Fusarium oxysporum f. sp. dianthi. The enzyme did not display activity towards hydroxycinnamic acid derivatives, some of which are involved, as methylated monolignols, in lignin biosynthesis; the role of this enzyme could be therefore mainly defensive, rather than structural, although its precise function still needs to be ascertained. PMID- 12899700 TI - Subunit sequences of the 4 x 6-mer hemocyanin from the golden orb-web spider, Nephila inaurata. AB - The transport of oxygen in the hemolymph of many arthropod and mollusc species is mediated by large copper-proteins that are referred to as hemocyanins. Arthropod hemocyanins are composed of hexamers and oligomers of hexamers. Arachnid hemocyanins usually form 4 x 6-mers consisting of seven distinct subunit types (termed a-g), although in some spider taxa deviations from this standard scheme have been observed. Applying immunological and electrophoretic methods, six distinct hemocyanin subunits were identified in the red-legged golden orb-web spider Nephila inaurata madagascariensis (Araneae: Tetragnathidae). The complete cDNA sequences of six subunits were obtained that corresponded to a-, b-, d-, e-, f- and g-type subunits. No evidence for a c-type subunit was found in this species. The inclusion of the N. inaurata hemocyanins in a multiple alignment of the arthropod hemocyanins and the application of the Bayesian method of phylogenetic inference allow, for the first time, a solid reconstruction of the intramolecular evolution of the chelicerate hemocyanin subunits. The branch leading to subunit a diverged first, followed by the common branch of the dimer forming b and c subunits, while subunits d and f, as well as subunits e and g form common branches. Assuming a clock-like evolution of the chelicerate hemocyanins, a timescale for the evolution of the Chelicerata was obtained that agrees with the fossil record. PMID- 12899701 TI - Structural characterization of the lipopolysaccharide O-polysaccharide antigen produced by Flavobacterium columnare ATCC 43622. AB - The structure of the antigenic O-chain polysaccharide of Flavobacterium columnare ATCC 43622, a Gram-negative bacterium that causes columnaris disease in warm water fish, was determined by high-field 1D and 2D NMR techniques, MS, and chemical analyses. The O-chain was shown to be an unbranched linear polymer of a trisaccharide repeating unit composed of 2-acetamido-2-deoxy-d-glucuronic acid (d GlcNAcA), 2-acetamidino-2,6-dideoxy-l-galactose (l-FucNAm) and 2-acetamido-2,6 dideoxy-d-xylo-hexos-4-ulose (d-Sug) (1 : 1 : 1), having the structure: [structure: see text]. PMID- 12899702 TI - Purification and characterization of NTPDase1 (ecto-apyrase) and NTPDase2 (ecto ATPase) from porcine brain cortex synaptosomes. AB - We purified to homogeneity and characterized NTPDase1 and NTPDase2 from porcine brain cortex synaptosomes. SDS/PAGE and immunoblotting with antibodies specific to these enzymes revealed a molecular mass estimated at 72 kDa for NTPDase1 and 66 for NTPDase2. Both enzymes exhibited kinetic properties typical for all members of the NTPDase family, e.g. low substrate specificity for tri- and diphosphonucleosides, divalent cations dependency and insensitivity towards ATPase inhibitors. The calculated Km value for NTPDase1 in respect to ATP as a substrate (97 microm) was three times lower in comparison to analogous values for NTPDase2 (270 microm). Additionally, NTPDase1 had a three times higher Kcat/Km coefficient than NTPDase2 (860 and 833 micromol product.s(-1), respectively). We have also demonstrated that in spite of differences in the affinity of ATP for both hydrolases, these enzymes have similar molecular activity. Taken together, these results indicate that NTPDase1 would terminate P2 receptor-mediated signal transmission whereas activity of NTPDase2 may contribute to decreasing high (toxic) concentrations of ATP and/or to production of another signal molecule, ADP. PMID- 12899703 TI - Purification, characterization, cDNA cloning and nucleotide sequencing of a cellulase from the yellow-spotted longicorn beetle, Psacothea hilaris. AB - A cellulase (endo-beta-1,4-glucanase, EC 3.2.1.4) was purified from the gut of larvae of the yellow-spotted longicorn beetle Psacothea hilaris by acetone precipitation and elution from gels after native PAGE and SDS/PAGE with activity staining. The purified protein formed a single band, and the molecular mass was estimated to be 47 kDa. The purified cellulase degraded carboxymethylcellulose (CMC), insoluble cello-oligosaccharide (average degree of polymerization 34) and soluble cello-oligosaccharides longer than cellotriose, but not crystalline cellulose or cellobiose. The specific activity of the cellulase against CMC was 150 micro mol.min-1.(mg protein)-1. TLC analysis showed that the cellulase produces cellotriose and cellobiose from insoluble cello-oligosaccharides. However, a glucose assay linked with glucose oxidase detected a small amount of glucose, with a productivity of 0.072 micro mol.min-1.(mg protein)-1. The optimal pH of P. hilaris cellulase was 5.5, close to the pH in the midgut of P. hilaris larvae. The N-terminal amino-acid sequence of the purified P. hilaris cellulase was determined and a degenerate primer designed, which enabled a 975-bp cDNA clone containing a typical polyadenylation signal to be obtained by PCR and sequencing. The deduced amino-acid sequence of P. hilaris cellulase showed high homology to members of glycosyl hydrolase family 5 subfamily 2, and, in addition, a signature sequence for family 5 was found. Thus, this is the first report of a family 5 cellulase from arthropods. PMID- 12899704 TI - Identification of sodium salicylate as an hsp inducer using a simple screening system for stress response modulators in mammalian cells. AB - As heat shock proteins (Hsps) are involved in protecting cells and also in the pathophysiology of diseases such as inflammation, cancer and neurodegenerative disorders, modulators of Hsp expression in mammalian cells would seem to be useful for the treatment of various diseases. In this study, we isolated mammalian cell lines for screening of Hsp modulators; mouse C3H10T1/2 cells stably transfected with a plasmid containing the mouse Hsp105 or human Hsp70B promoter upstream of a luciferase or beta-galactosidase reporter gene, respectively. Using these cells, we examined the effect of sodium salicylate (SA), which may induce the transcription of hsp genes, on stress response in mammalian cells. When these cells were treated with SA for 1 h at 37 degrees C, both promoter activities were up-regulated by SA at concentrations of more than 45 mm. The activation of heat shock factor and the subsequent accumulation of Hsp105alpha and Hsp70 were detected in cells treated with SA at concentrations of more than 20 and 45 mm, respectively. Furthermore, SA induced resistance against a subsequent lethal stress. These findings suggested that SA is a potent hsp inducer, and may be used to protect cells against deleterious stressors. PMID- 12899705 TI - Cause of mortality in insects under severe stress. AB - Mortality in the host armyworm larvae Pseudaletia separata parasitized by the parasitic wasp Cotesia kariyai was dramatically increased when they were simultaneously infected by the entomopathogen Serratia marcescens. Previous studies have shown that this strong insecticidal effect is due to a metalloprotease-like insecticide (MPLI) released from S. marcescens enterobacter. This study was conducted to elucidate the exact cause of the mortality resulting from MPLI. Injection of MPLI caused a sharp increase in hemolymph dopamine concentration followed by elevated levels of brain dopamine in armyworm larvae. [3H]Dopamine injected into the hemocoel, was incorporated into the brains of MPLI injected larvae to a level eight times greater than in BSA-injected control larvae. Transmission electron microscopy showed an obvious decrease in thickness and density of the brain sheath in insects injected with MPLI. This was probably due to the MPLI-induced elevation of hemocyte metalloprotease activities. Further, electron microscopic and TUNEL staining analyses showed a significant increase in apoptotic cells in the brain 12 h after the injection. Injection of 3 iodotyrosine (a tyrosine hydroxylase inhibitor) before MPLI completely prevented the increase in hemolymph dopamine in test larvae and their following death. From these observations, we conclude that MPLI-injected larvae may have suffered mortal damage through increased apoptosis of brain cells caused by an influx of dopamine from the hemolymph. PMID- 12899706 TI - Fanconi's anaemia. PMID- 12899707 TI - Wolman's disease in an infant. PMID- 12899708 TI - Flt3 mutations and leukaemia. PMID- 12899709 TI - The history of acute promyelocytic leukaemia. PMID- 12899710 TI - Early history of iron deficiency. PMID- 12899711 TI - Acute promyelocytic leukaemia in patients originating in Latin America is associated with an increased frequency of the bcr1 subtype of the PML/RARalpha fusion gene. AB - The PML/RARalpha fusion gene in acute promyelocytic leukaemia (APL) has three subtypes based on the breakpoint site of the PML gene: long (bcr1), short (bcr3) and variable (bcr2) subtypes. The PML/RARalpha fusion protein is involved in the pathogenesis of APL and the breakpoint site of the PML gene might be associated with aetiological factor(s). Because APL is over-represented in patients that originate in Latin America (Latinos), we evaluated whether the distribution of the PML/RARalpha fusion mRNA in this population is different to that reported in non-Latinos. Among 52 APL patients (28 from Mexico and Central America diagnosed in Los Angeles and 24 from Peru, South America), bcr1, bcr2 and bcr3 expression was 75%, 10% and 15% respectively. However, bcr1 breakpoints were significantly higher compared with non-Latino patients (340/654, 52%) reported in four studies. Often bcr1 and bcr2 are reported together; 862 (60%) of 1429 non-Latino APL patients reported in nine studies were either bcr1 or bcr2, compared with 44 (85%) in our 52 Latino patients. This difference was also statistically significant when our patients were compared to each of the individual studies from USA and Europe, but not for a small series from China and Japan. These results suggest that the overrepresentation of APL among Latin American patients can be accounted for by an increase of a single subtype--bcr1, and the breakage sites in the PML gene may not be random but possibly influenced by genetic and/or environmental factor(s). PMID- 12899712 TI - Aberrant gene promoter methylation in acute promyelocytic leukaemia: profile and prognostic significance. AB - Acute promyelocytic leukaemia (APL) has distinct clinicopathological and molecular features. However, the profile of aberrant gene promoter methylation is undefined. In this study, methylation-specific polymerase chain reaction (MSP) was used to define the methylation status of a panel of nine genes, comprising p15, p16, RARbeta, oestrogen receptor (ER), E-cadherin (E-CAD), p73, caspase 8 (CASP8), VHL and MGMT, in 29 patients with APL. Aberrant methylation of p15, ER, RARbeta, p16 and E-CAD occurred, respectively, in 23 (79%), 14 (48%), six (21%), six (21%) and two (7%) patients at diagnosis, but p73, VHL, CASP8 and MGMT were not methylated in any patients. There was methylation of one gene in 13 patients (45%), two genes in four patients (14%), three genes in six patients (21%) and four genes in three patients (10%). Concurrent methylation of two or more genes occurred in 13 patients (45%). No association was identified between gene methylation and presenting clinicopathological features. However, p15 methylation was significantly associated with an inferior disease-free survival (DFS, P = 0.008), and remained the only poor prognostic factor in multivariate analysis (P = 0.019). In APL, p15, p16, ER and RARbeta were most frequently methylated. This profile is distinct from other types of myeloid leukaemias. p15 methylation has a poor prognostic impact on DFS. PMID- 12899713 TI - CXCR4 chemokine receptors (CD184) and alpha4beta1 integrins mediate spontaneous migration of human CD34+ progenitors and acute myeloid leukaemia cells beneath marrow stromal cells (pseudoemperipolesis). AB - Marrow stromal cells play an important role in regulating the development and proliferation of haematopoietic stem cells (HSC) within the marrow microenvironment. However, the molecular mechanisms of stem cell-stromal cell interactions are not fully understood. We observed that mobilized peripheral blood and cord-blood-derived CD34+ progenitor cells, or CD34+ acute myeloid leukaemia (AML) cells spontaneously migrated beneath marrow stromal cells, an in vitro migration phenomenon termed pseudoemperipolesis. In contrast, the CD34+ myeloid leukaemia cell line, Kasumi-1, did not display pseudoemperipolesis. Cord blood CD34+ cells had a higher capacity than granulocyte-colony-stimulating factor-mobilized CD34+ cells for pseudoemperipolesis (28.7 +/- 12%vs 18.1 +/- 6.1% of input cells within 24 h, mean +/- SD, n = 8), whereas 9.4 +/- 12.6% (mean +/- SD, n = 10) of input AML cells displayed this phenomenon. Pseudoemperipolesis of CD34+ progenitor and AML cells was significantly inhibited by pertussis toxin and antibodies to the CXCR4 chemokine receptor (CXCR4, CD184), but not control antibodies. Moreover, CD34+ and AML cell migration was significantly inhibited by a CS1 peptide that blocks alpha4beta1 integrin binding, but not by a control peptide, in which the fibronectin binding motif was scrambled. Pseudoemperipolesis was associated with an increased proliferation of migrated CD34+ progenitor cells but not AML cells within the stromal layer, demonstrated by cell cycle analysis and cell division tracking. We conclude that alpha4beta1 integrin binding and CXCR4 chemokine receptor activation are prerequisites for the migration of CD34+ haematopoietic progenitors and AML cells beneath marrow stromal cells. These observations suggest a central role of marrow stromal cells for HSC trafficking and homing within the marrow microenvironment. PMID- 12899714 TI - A 2.6 Mb interval on chromosome 6q25.2-q25.3 is commonly deleted in human nasal natural killer/T-cell lymphoma. AB - Natural killer (NK)/T-cell lymphoma is a special subtype of rare malignant lymphoma that is more prevalent in Asia than in America and Europe. This newly characterized haemato-lymphoid malignancy is highly aggressive and frequently present in nasal and upper aerodigestive sites. Several studies have reported the commonly deleted region of chromosome 6q21-25 in this particular type of lymphoma. To refine the smallest region of overlapping (SRO) deletion for localization of potential tumour suppressor (TS) genes, we performed loss of heterozygosity (LOH) and homozygosity mapping of deletion (HOMOD) analyses on 37 nasal and nasal-type NK/T-cell lymphoma patients using a panel of 25 microsatellite markers, covering the 6q21-q25 region. In all patients studied, LOH was detected in eight (89%) paired-sample patients, while hemizygous deletion was detected in three (11%) single-sample patients. Combination of the LOH and HOMOD results defined a distinct 3 Mb SRO on chromosome 6q25. Quantitative multiplex polymerase chain reaction analysis of 10 sequence-tagged sites further refined the putative TS-gene-containing region to a 2.6 Mb interval between TIAM2 and SNX9. Eighteen known genes/Unigene clusters and 25 hypothetical genes are located within this 2.6 Mb region, but none are previously identified TS genes. These results provide a framework for future positional cloning of novel TS gene(s) at 6q25.2-q25.3. PMID- 12899715 TI - Disease activity and pretreatment, rather than hypogammaglobulinaemia, are major risk factors for infectious complications in patients with chronic lymphocytic leukaemia. AB - To identify patients at high risk of life-threatening infections, we retrospectively analysed the prevalence of infectious complications in 187 chronic lymphocytic leukaemia patients treated in our institution since 1999 and correlated them with clinical features. A questionnaire with detailed questions regarding infectious complications was mailed to patients and their general practitioners. Major infections (requiring intravenous antibiotics or inpatient treatment) were reported in 37 patients (19.8%) and minor infections (requiring oral antibiotics and outpatient treatment) in 113 patients (60.4%). Univariate analysis identified advanced disease (P = 0.02), gender (P = 0.01), duration of disease (P = 0.007), number of previous chemotherapy regimens (P < 0.001), previous therapy with purine analogues and monoclonal antibodies (P < 0.001; P = 0.019), massive splenomegaly (P = 0.03), low granulocyte count (P < 0.001), low serum immunoglobulin concentration (P = 0.005), low haemoglobin concentration (P < 0.001) and high serum lactate dehydrogenase (LDH) concentration (P < 0.001) as risk factors for major infections. In multivariable logistic regression analysis, only the number of previous chemotherapy regimens (risk ratio [RR] = 1.8; 95% confidence interval [CI] 1.2-8.0) and haemoglobin concentration (RR = 0.6; CI 0.5 0.8) remained significant for major infections. The number of previous chemotherapy regimens was the only independent risk factor for minor (RR = 7.6; CI 2.2-25.7) and varicella-zoster virus infections (RR = 2.1; CI 1.3-3.4). In untreated patients, the only risk factor for major infections was LDH concentration. Patients treated with purine analogues or autologous stem cell transplantation had a higher risk of developing viral infections. In conclusion, disease activity and pretreatment extent have a stronger impact on the risk of severe infectious complications than hypogammaglobulinaemia. Preferably, prophylactic strategies should be evaluated in patients defined by these parameters. PMID- 12899716 TI - Hyperfractionated cyclophosphamide in combination with pulsed dexamethasone and thalidomide (HyperCDT) in primary refractory or relapsed multiple myeloma. AB - Sixty patients with advanced multiple myeloma received 2-6 monthly treatment courses combining hyperfractionated cyclophosphamide (300 mg/m2 i.v. over 3 h q 12 h x 6, d 1-3) with pulsed dexamethasone (20 mg/m2/d p.o., d 1-4, 9-12, 17-20) and once daily thalidomide at individually escalating doses (100-400 mg/d) depending on tolerability (HyperCDT). Responding patients were maintained on daily thalidomide and monthly dexamethasone pulses. Complete, partial and minor response rates were 4%, 68% and 12% respectively; overall response rate was 84% (efficacy analysis). Median event-free and overall survival was 11 and 19 months respectively. During at least one treatment cycle, 67% of patients experienced grade 4 neutropenia resulting in 17% grade 3 and 9% grade 4 infections. Side effects, presumably related to thalidomide, included neuropathy (40% grade 2, 16% grade 3), constipation (17%), oedema (5%), bradycardia (5%), skin reactions (3%), cerebrovascular events (5%) and deep vein thromboses (8%). Thromboses were not related to known thrombophilic risk factors. Four patients with prior myeloma therapy > 50 months developed myelodysplastic syndrome or secondary acute myeloid leukaemia 2-4 months after study entry. HyperCDT is a highly active and reasonably well-tolerated salvage regimen in advanced or refractory multiple myeloma. PMID- 12899717 TI - Severe and selective deficiency of interferon-gamma-producing invariant natural killer T cells in patients with myelodysplastic syndromes. AB - Here we show that patients with myelodysplastic syndromes (MDS) have a severe deficiency of glycolipid reactive Valpha24+/Vbeta11+ natural killer T (NKT) cells, but not NK cells or CD4+ or CD8+ T cells. Neither the blood nor marrow of MDS patients had detectable interferon-gamma-producing NKT cells in response to the NKT ligand, alpha-galactosyl ceramide, although influenza-virus-specific effector T-cell function was preserved. This severe and selective deficiency of an important immune regulatory cell may contribute to the pathogenesis of MDS. PMID- 12899718 TI - Kinetics of haematopoietic recovery after dose-intensive chemo/radiotherapy in mice: optimized erythroid support with darbepoetin alpha. AB - Despite its frequency and impact on clinical outcomes, anaemia in cancer patients remains poorly understood and suboptimally treated. The definition of optimum treatment schedules with erythropoietic agents requires a suitable model of chemotherapy-induced progressive anaemia. This study investigated novel strategies such as once-per-chemotherapy-cycle dosing, synchronization between erythroid supportive care and chemotherapy, and definition of the optimum timing of erythroid support. A murine model of carboplatin chemotherapy/radiotherapy (CRT)-induced anaemia was used, which caused progressive anaemia across multiple cycles. Weekly administration of recombinant human erythropoietin (rHuEPO) was effective, but the longer-acting darbepoetin alpha resulted in superior responses. In all animals, anaemia became progressive and more refractory across cycles because of accumulated bone marrow damage. Exploiting a specific enzyme linked immunosorbent assay, which could distinguish between darbepoetin alpha and endogenous erythropoietin, the effect of CRT upon the pharmacokinetics of darbepoetin alpha showed that clearance of darbepoetin alpha, and presumably erythropoietin, was at least partially dependent on a chemotherapy-sensitive pathway. Scheduling data suggested that administration of erythropoietic agents prior to chemotherapy was more effective than administration after chemotherapy. There was no evidence that erythropoietic agents exacerbated anaemia, even when administered immediately prior to CRT in an attempt to "prime" erythroid cells for the effects of CRT. PMID- 12899719 TI - Graft-versus-host disease (GVHD)-specific survival and duration of systemic immunosuppressive treatment in patients who developed chronic GVHD following allogeneic haematopoietic cell transplantation. AB - We investigated graft-versus-host disease (GVHD)-specific survival (GSS) and the duration of systemic immunosuppressive treatment (IST) in 82 patients who developed chronic GVHD following allogeneic haematopoietic cell transplantation (HCT). These two major study endpoints were calculated using the Kaplan-Meier method. Deaths solely due to the relapse of underlying disease or accidental deaths were censored at the time of occurrence for the analysis of GSS. The probability of GSS at 5 years was 74.2%. The median duration of systemic IST for chronic GVHD was 272 d (range: 7-1450), and the probability of withdrawal of systemic IST at 1, 2 and 3 years was 67.3%, 82.4% and 89.0% respectively. Analysis based on a multivariate model showed that a diagnosis other than leukaemia or myelodysplastic syndrome (P = 0.049), prior occurrence of grade III IV acute GVHD (P = 0.021), onset of chronic GVHD before d 120 (P = 0.013), serum alkaline phosphatase over 120 IU/l (P = 0.034), and serum bilirubin over 34.2 micromol/l (P = 0.015) were independent adverse prognostic factors for GSS. Prior occurrence of grade III-IV acute GVHD significantly influenced the duration of systemic IST (P = 0.048). In conclusion, analyses of GSS and the duration of systemic IST will allow patients with different outcomes to be stratified for appropriate treatment application and will provide important parameters in prospective trials for the treatment of chronic GVHD. PMID- 12899720 TI - Does sticky blood predict a sticky end? Associations of blood viscosity, haematocrit and fibrinogen with mortality in the West of Scotland. AB - There is increasing evidence that blood viscosity and its major determinants (haematocrit, plasma viscosity and fibrinogen) are associated with an increased risk of incident cardiovascular events; however, their associations with mortality are not established. We therefore studied the associations of these variables with cardiovascular events and total mortality in 1238 men and women aged 25-64 years, followed for 13 years in the first North Glasgow MONICA (MONItoring CArdiovascular disease) survey and West of Scotland centres in the Scottish Heart Health Study. After adjustment for age and sex, increasing whole blood viscosity, plasma viscosity, haematocrit and fibrinogen (analysed by both von Clauss and heat precipitation assays) were significantly associated with mortality. Only the association for fibrinogen (von Clauss assay) remained significant after adjustment for major cardiovascular risk factors. We conclude that clottable fibrinogen may be independently associated with mortality. However, the significance of this association, and the extent to which viscosity is associated with mortality, remain to be established in larger studies and meta analyses. PMID- 12899721 TI - Activation of platelets in whole blood by recombinant factor VIIa by a thrombin dependent mechanism. AB - Using a diluted whole blood method of flow cytometric analysis, we have shown that platelets could be activated in vitro in the presence of high concentrations (100 nmol/l) of recombinant factor (F) VIIa (rFVIIa; NovoSeven(R)) and 2.5 mmol/l calcium chloride. This was demonstrated by a significant increase in the mean percentage of platelets expressing CD62P and their mean fluorescent intensity (MFI) after 30 min versus platelets incubated with calcium or rFVIIa alone or diluted blood alone. The presence of rFVIIa and calcium increased the exposure of the PAC-1 activation epitope of glycoprotein (Gp) IIb/IIIa. This effect was equally influenced by the presence of calcium alone but not by rFVIIa. The effect of rFVIIa was time and concentration dependent. Thrombin generation was also necessary, as the effect of rFVIIa was completely abrogated by the additional presence of hirudin. Furthermore, soy bean trypsin inhibitor (SBTI) but not corn trypsin inhibitor (CTI) abrogated CD62P exposure, suggesting that thrombin was derived via FX but not FXII activation. Exposure of CD62P demonstrated a significant lag phase, sometimes of the order of > 30 min, as well as large intersubject variation. Significant platelet activation was observed at a concentration as low as 25 nmol/l rFVIIa. Platelet-leucocyte aggregation was also increased in the presence of 25 nmol/l rFVIIa and calcium. No significant difference was observed between levels of CD62P in diluted whole blood and platelet-rich plasma adjusted to an identical platelet count after their exposure to rFVIIa and calcium for 30 min. PMID- 12899722 TI - Expression of Plasmodium falciparum G6PD-6PGL in laboratory parasites and in patient isolates in G6PD-deficient and normal Nigerian children. AB - As the production of NADPH in the pentose phosphate pathway is the main antioxidant defence mechanism available to the Plasmodium falciparum, we have studied the expression of P. falciparum glucose 6-phosphate dehydrogenase-6 phosphogluconolactonase (PfG6PD-6PGL) in G6PD-deficient and normal erythrocyte host cells. Both erythrocytes infected in vitro with a laboratory isolate and erythrocytes from natural human infections were used. Total RNA was prepared from parasites collected from five G6PD-deficient and nine G6PD-normal children in Ibadan, Nigeria, selected after screening 189 rural schoolchildren and 68 clinical malaria patients, and was subjected to Northern blot analysis. The probe was a cDNA fragment of the G6PD domain of the PfG6PD-6PGL gene, with an internal control probe of P. falciparum 18S ribosomal RNA. Quantification was performed using a phosphoimager. Relative to internal control, the abundance of PfG6PD-6PGL mRNA (mean +/- standard deviation) was lower in parasites from G6PD-deficient children (0.29 +/- 0.27) than in G6PD-normal control subjects (0.74 +/- 0.26) (P = 0.014, Mann-Whitney U-test). Although confirmation in a larger study is required, our results suggest a lower relative abundance of PfG6PD-6PGL, and presumably antioxidant activity, in malaria parasites from G6PD-deficient hosts, thus extending the current knowledge of the mechanism of G6PD-deficiency related host protection. PMID- 12899723 TI - Simultaneous (AC)n microsatellite polymorphism analysis and single-stranded conformation polymorphism screening is an efficient strategy for detecting ankyrin-1 mutations in dominant hereditary spherocytosis. AB - Nonsense/stop mutations in the ankyrin-1 gene (ANK1) are a major cause of dominant HS (dHS) (frequency of 23% in German dHS patients). To date, no common mutation has been found and therefore a simple mutation screening is not feasible. The reduced expression of one cDNA allele in the (AC)n microsatellite polymorphism of the ankyrin-1 gene, as seen in about 20% of Czech patients with dHS, may identify candidates with a possible frameshift/nonsense mutation. In order to verify the efficiency of this screening we screened the ankyrin-1 gene of 22 Czech dHS patients for both the reduced cDNA allele expression in the frequent (AC)n and the common exonic 26/39 polymorphisms, as well as for polymerase chain reaction (PCR) single-stranded conformation polymorphisms in any one of the 42 exons of ANK1. Anomalous PCR products were sequenced. We found seven new ANK1 frameshift/nonsense mutations in nine patients with, but in none of six patients without, a reduced cDNA allele expression (efficiency of 78%). We conclude that screening of dHS patients for such a reduced allele expression in common ANK1 polymorphisms is an efficient procedure for the identification of candidates for frameshift/nonsense mutations in the ankyrin-1 gene. PMID- 12899724 TI - Increasing oxygen tension improves filtration of sickle trait donor blood. AB - A major cause of filter failure of red cell (RBC) components from donors with sickle cell trait (HbAS) is the polymerization of haemoglobin. The oxygen saturation (sO2) of blood stored in various plastics and different volumes of air was assessed. Blood from 10 HbAS donors was collected and divided into two bags, one with air added, one without. Bags with added air had increased sO2 levels (from 49 +/- 10% to 76 +/- 6%). Filtration was successful for nine of 10 components with air, and one of 10 without air. Successful filtration of RBC components occurs when sO2 is increased. PMID- 12899725 TI - McLeod syndrome resulting from a novel XK mutation. AB - McLeod Syndrome (MLS) is a rare X-linked disorder characterized by haemopoietic abnormalities and late-onset neurological and muscular defects. The McLeod blood group phenotype is typically associated with erythrocyte acanthocytosis, absence of the Kx antigen and reduced expression of Kell system antigens. MLS is caused by hemizygosity for mutations in the XK gene. We describe a patient with MLS who first showed symptoms in 1989 (aged 51 years). As the disease progressed, the patient developed a slight dementia, aggressive behaviour and choreatic movements. A cardiomyopathy was also diagnosed. An electroneuromyography showed neuropathic and myopathic changes. Liver enzymes were elevated and a blood smear showed acanthocytes. MLS was confirmed by serological analysis of the Kell antigens. Analysis of red blood cells by flow cytometry revealed the patient and his grandson to have reduced Kell antigen expression. The patient's daughters had two populations of red cells, consistent with them being heterozygous for an XK0 allele. The molecular basis of MLS in this family is a novel mutation consisting of a 7453-bp deletion that includes exon 2 of the XK gene. This confirms that the patient's 7-year-old grandson, who is currently asymptomatic, also has the XK0 allele and is therefore likely to develop MLS. PMID- 12899726 TI - Transient leukaemia in newborn infants with trisomy 21. PMID- 12899727 TI - Successful use of gemtuzumab ozogamicin in a child with relapsed CD33-positive acute lymphoblastic leukaemia. PMID- 12899728 TI - Isolated epidural chloroma with translocation t(15; 17) successfully treated with chemotherapy and all-trans-retinoic acid. PMID- 12899729 TI - Rituximab effective in a patient with refractory autoimmune haemolytic anaemia and CD20-negative multiple myeloma. PMID- 12899731 TI - Dual effects of nicotine on dopamine neurons mediated by different nicotinic receptor subtypes. AB - Burst firing of dopaminergic neurons has been found to represent a particularly effective means of increasing dopamine release in terminal areas as well as activating immediate early genes in dopaminoceptive cells. Spontaneous burst firing is largely controlled by the level of activation of NMDA receptors in the ventral tegmental area (VTA) as a consequence of glutamate released from afferents arising mainly in the prefrontal cortex. Nicotine has been found to effectively increase burst firing of dopaminergic cells. This effect of nicotine may be due to an alpha 7 nicotinic receptor-mediated presynaptic facilitation of glutamate release in the VTA. By the use of in-vivo single-cell recordings and immunohistochemistry we here evaluated the role of alpha 7 nicotinic receptors in nicotine-induced burst firing of dopamine cells in the VTA and the subsequent activation of immediate early genes in dopaminoceptive target areas. Nicotine (0.5 mg/kg s.c.) was found to increase firing rate and burst firing of dopaminergic neurons. In the presence of methyllycaconitine (MLA, 6.0 mg/kg i.p.) nicotine only increased firing rate. Moreover, in the presence of dihydro-beta erythroidine (DH beta E, 1.0 mg/kg i.p.), an antagonist at non-alpha 7 nicotinic receptors, nicotine produced an increase in burst firing without increasing the firing rate. Nicotine also increased Fos-like immunoreactivity in dopamine target areas, an effect that was antagonized with MLA but not with DH beta E. Our data suggest that nicotine's augmenting effect on burst firing is, indeed, due to stimulation of alpha 7 nicotinic receptors whereas other nicotinic receptors seem to induce an increase in firing frequency. PMID- 12899730 TI - Intermediate filament-co-localized molecules with myosin heavy chain epitopes define distinct cellular domains in hair follicles and epidermis. AB - BACKGROUND: Proteins linking intermediate filaments to other cytoskeletal components have important functions in maintaining tissue integrity and cell shape. RESULTS: We found a set of monoclonal antibodies raised against specific human sarcomeric myosin heavy chain (MyHC) isoforms labels cells in distinct regions of the mammalian epidermis. The antigens co-localize with intermediate filament-containing structures. A slow MyHC-related antigen is punctate on the cell surface and co-localizes with desmoplakin at desmosomal junctions of all suprabasal epidermal layers from rat foetal day 16 onwards, in the root sheath of the hair follicle and in intercalated disks of cardiomyocytes. A fast MyHC related antigen occurs in cytoplasmic filaments in a subset of basal cells of skin epidermis and bulb, but not neck, of hair follicles. A fast IIA MyHC-related antigen labels filaments of a single layer of cells in hair bulb. This 230,000 Mr antigen co-purifies with keratin. No obvious candidate for any of the antigens appears in the literature. CONCLUSIONS: We describe a set of molecules that co localize with intermediate filament in specific cell subsets in epithelial tissues. These antigens presumably influence intermediate filament structure or function. PMID- 12899732 TI - Enhancing action of LSD on neuronal responsiveness to serotonin in a brain structure involved in obsessive-compulsive disorder. AB - Potent serotonin (5-HT) reuptake inhibitors are the only drugs that consistently exert a therapeutic action in obsessive-compulsive disorder (OCD). Given that some hallucinogens were reported to exert an anti-OCD effect outlasting their psychotomimetic action, possible modifications of neuronal responsiveness to 5-HT by LSD were examined in two rat brain structures: one associated with OCD, the orbitofrontal cortex (OFC), and another linked to depression, the hippocampus. The effects of concurrent microiontophoretic application of LSD and 5-HT were examined on neuronal firing rate in the rat OFC and hippocampus under chloral hydrate anaesthesia. In order to determine whether LSD could also exert a modification of 5-HT neuronal responsiveness upon systemic administration, after a delay when hallucinosis is presumably no longer present, it was given once daily (100 microg/kg i.p.) for 4 d and the experiments were carried out 24 h after the last dose. LSD attenuated the firing activity of OFC neurons, and enhanced the inhibitory effect of 5-HT when concomitantly ejected on the same neurons. In the hippocampus, LSD also decreased firing rate by itself but decreased the inhibitory action of 5-HT. The inhibitory action of 5-HT was significantly greater in the OFC, but smaller in the hippocampus, when examined after subacute systemic administration of LSD. It is postulated that some hallucinogens could have a beneficial action in OCD by enhancing the responsiveness to 5-HT in the OFC, and not necessarily in direct relation to hallucinosis. The latter observation may have theoretical implications for the pharmacotherapy of OCD. PMID- 12899733 TI - Tolerance to the anhedonic effects of lipopolysaccharide is associated with changes in syntaxin immunoreactivity in the nucleus accumbens. AB - The bacterial endotoxin lipopolysaccharide (LPS) produces a host of effects in mammals known collectively as 'sickness' behaviours. Acute treatment with LPS also results in a loss of hedonic capacity in rodents that can be measured by changes in responding for reinforcing electrical stimulation of the lateral hypothalamus. In contrast, repeated exposure to LPS typically leads to the development of tolerance to many of the physiological and behavioural effects of endotoxin, although the effect of chronic treatment with LPS on anhedonia remains unknown. In the present experiment, rats were trained to respond on an ascending series current-intensity intracranial self-stimulation (ICSS) protocol, and were then treated with either acute or sub-chronic LPS (100 microg). Compared to vehicle-treated subjects, acute exposure to LPS induced a dramatic loss of ICSS responding; however, with repeated exposure to LPS, rats developed a behavioural tolerance to its anhedonic effects. To investigate a potential molecular substrate for the anhedonic effects of LPS, quantitative immunohistochemistry was used to measure levels of the synaptic proteins syntaxin, SNAP-25 and synaptophysin in the dorsal and ventral striatum of rats treated acutely and sub chronically with LPS. A single injection of LPS produced a significant decrease in syntaxin immunoreactivity in the nucleus accumbens core and shell, while similar treatment in chronically treated rats that displayed behavioural tolerance had no effect. These results demonstrate a novel molecular substrate for the effects of LPS, and imply that the underlying physiology of the transient anhedonic effects of LPS may differ from that involved in chronic psychiatric disorders in humans. PMID- 12899734 TI - N-methyl-norsalsolinol modulates serotonin metabolism in the rat caudate nucleus: correlation with behavioural changes. AB - In earlier studies the dihydroxylated tetrahydroisoquinoline derivative 2(N) methyl-norsalsolinol (NMNorsal) was identified in patients with Parkinson's disease. In the present study, NMNorsal (20 or 40 mg/kg) was given intraperitoneally to rats kept under normal light-dark cycles. Using brain microdialysis technique, serotonin (5-HT), 5-hydroxyindolacetic acid (HIAA), dopamine (DA), and 3,4-dihydroxyphenylacetic acid (DOPAC) were determined in the dialysate from caudate nucleus in vivo and from tissue in vitro at various times following NMNorsal administration. Even after high-dose NMNorsal administration (40 mg/kg) and measurements up to 48 h after administration, levels of DA and its metabolite DOPAC were not modified. In contrast to the DA metabolism, 5-HT levels in the dialysate increased to approx. 2-fold during the 48 h following administration of a single high-dose of NMNorsal while HIAA decreased to approx. 50%. These changes of 5-HT and HIAA were nearly identical in the homogenate preparation of the caudate nucleus when compared to the amounts present in the dialysate. During assessment controls and low-dose-treated animals were almost always sleeping. Only high-dose NMNorsal-treated rats were active, with maximum activity after 48 h, however, behavioural activity was clearly different to the classical 5-HT behavioural syndrome. Taken together, increased 5-HT levels in the striatum found in our studies seem to be linked to the behavioural activity induced by high-dose NMNorsal, and NMNorsal appeared to perturb normal diurnal rhythms of spontaneous locomotor activity. The precise mechanism by which NMNorsal acts on 5-HT metabolism and behaviour is, however, unclear and further investigation is required. PMID- 12899735 TI - Algorithm-based treatment of major depression in an outpatient clinic: clinical correlates of response to a specific serotonin reuptake inhibitor and to triiodothyronine augmentation. AB - Clinicians who treat major depression are faced with a bewildering choice of antidepressants. Given that all have a lag period before they are effective, choice of the right antidepressant can potentially minimize the duration of symptoms, reduce cost and enhance compliance. Unfortunately, there is very little evidence upon which to base such a decision. We developed a treatment algorithm for non-psychotic, unipolar major depression, and applied it in our clinic. It includes progression from a specific serotonin reuptake inhibitor (SSRI), usually fluoxetine, from a dose of 20 mg to a dose of 40 mg in patients who do not respond. Non-responders to this dose receive augmentation with triiodothyronine (T3, 25-50 microg). All interventions are for fixed time periods and guided by overall clinical improvement as defined by the Clinical Global Inventory. Ninety patients commenced open-label treatment with 20 mg SSRI (fluoxetine, n=81; paroxetine, n=9). Seventy-four patients completed 4 wk of treatment and 44 (48.9%) were responders (intent to treat analysis). Raising the SSRI dose to 40 mg for a further 2 wk was effective in only 5 patients (16.6%). Non-responders to SSRI were significantly more depressed at baseline as reflected by their rating scales scores. Addition of T3 was effective in 10 out of 16 women (62.5%) while none of the 9 males responded. Although values were within the normal range, patients who responded to T3 had higher serum thyroid-stimulating hormone levels than those who did not. Our experience with algorithm-based treatment of unipolar, non-psychotic major depression in outpatients suggests that more than 40% of patients will not respond to initial treatment with an SSRI even when the dose is increased to 40 mg/d; that severity of depression may be an important predictor of response and that T3 may be useful as an augmenter of response in SSRI non-responders but may be less effective in men than in women. The effect of T3 may be related to thyroid function even within the normal range. PMID- 12899736 TI - Serotonin depletion increases respiratory variability in freely moving rats: implications for panic disorder. AB - To elucidate if serotonergic transmission affects respiratory variability, a parameter consistently found increased in patients with panic disorder, we studied the effect of a serotonin synthesis inhibitor, para-chlorophenylalanine (PCPA), on respiratory variability at baseline and during CO2-induced hyperventilation in awake and unrestrained rats. Forty male Wistar rats were given intraperitoneal injections of PCPA (300 mg/kg) or saline 72, 48 and 24 h before registration of respiration in a plethysmograph allowing the animals to move freely. PCPA-treated rats displayed significantly higher tidal volume variability and minute volume variability, both at baseline and during CO2 exposure, compared to controls. The results support the notion that serotonin dysfunction may contribute to the enhanced respiratory variability observed in patients with panic disorder. PMID- 12899737 TI - Glucuronidation enzymes, genes and psychiatry. AB - The phase I cytochrome P450 (CYP) isoenzymes have received substantial attention in the pharmacogenetic literature. Researchers are beginning to examine the role of the phase II UDP-glucuronosyltransferase (UGT) enzymes, which produce products that are more water-soluble, less toxic and more readily excreted than the parent compounds. Several reasons may have contributed to neglect of UGTs (compared to CYPs) including: (1) the overlapping activity of UGTs and lack of selective probes; (2) the complexity of the glucuronidation cycle; and (3) the difficulty in developing analytic methods to measure glucuronides. Current CYP knowledge is used as a model to predict advances in UGT knowledge. At least 24 different UGT human genes have been identified and are classified in two families (UGT1 and UGT2) based on sequence homology. The UGT1A subfamily (genes located on chromosome 2) glucuronidates bilirubin, thyroid hormones, and some medications. UGT1A4 metabolizes tricyclic antidepressants and some antipsychotics. The UGT2B subfamily (genes located on chromosome 6) glucuronidates sexual steroids and bile acids. Oxazepam and lorazepam are mainly metabolized by glucuronidation. Anti epileptics with mood-stabilizing properties are frequently metabolized by UGTs. Opioid and nicotine addiction may also be influenced by glucuronidation. Glucuronidation of serotonin may be important during fetal development. UGTs appear to be in small concentrations in brain tissue (and higher concentrations at brain capillaries). However, UGTs may be localized in certain brain areas to provide a neuroprotective function. This review illustrates the importance of glucuronidation and the implications for psychiatry. PMID- 12899738 TI - Vagus nerve stimulation (VNS): utility in neuropsychiatric disorders. AB - Early in the last century, neuroscientists initially speculated that it might be possible to influence brain activity and associated behaviours by stimulating the vagus nerve in the neck. Several animal studies pointed to this possibility. In the 1980s, Zabara discovered that vagus nerve stimulation (VNS) could reduce or eliminate seizures in dogs. His discovery led to enhanced methods of direct VNS and the development of an implantable device for human VNS. VNS is now an accepted long-term treatment for epilepsy, used by over 15000 patients worldwide. Many have wondered whether VNS could treat other neuropsychiatric disorders. This paper reviews the neurobiology involved in VNS, highlights the VNS clinical findings to date, and discusses potential VNS research opportunities for both clinical and basic investigators. Comments on the clinical utility of VNS in actual clinical practice are provided. PMID- 12899739 TI - Fluvoxamine-induced hyperglycaemia in a diabetic patient with comorbid depression. PMID- 12899740 TI - Mania associated with venlafaxine discontinuation. PMID- 12899741 TI - Another marker for different types of depression. PMID- 12899744 TI - [Clinical analysis of 190 cases of outbreak with atypical pneumonia in Guangzhou in spring, 2003]. AB - OBJECTIVE: To study methods of diagnosis and treatment for atypical pneumonia (Severe Acute Respiratory Syndrome), outbreak of the illness in Guangzhou during Jan. - Mar., 2003. METHODS: 190 cases with atypical pneumonia were analyzed, and the cases were admitted in Guangzhou municipal First Hospital and Guangzhou municipal Eighth Hospital. RESULTS: Patients were infected by close quarters contacting each other. All patients manifest high fever, and accompany by dyspnea, cough, palpitate, weakness, headache and swirl. Other 46 cases were accompanied by diarrhea. Most of patients, manifestation of lungs was negative. Chest X-ray, shadow of lungs were light in beginning, and change to severity slowly or suddenly during 5 - 10 days. Of these cases, 36 cases develop to ARDS and 11 cases died with severity ARDS. Using general antibiotic was of no effect for the illness. Continual positive airway pressure (CPAP) and glucocorticoid was required that can control deprivation of the disease when toxicosis symptom of patients was severity and shadow of lungs diffuse more and more. CONCLUSION: Infectivity of the illness is evidence and spread by airway. Using general antibiotic was of no effect for the illness. Continual positive airway pressure (CPAP) and glucocorticoid are effective for control of the disease. PMID- 12899745 TI - [Interaction of Helicobacter polyri and indomethacin in gastric mucosa injury of Balb/c mice]. AB - OBJECTIVE: To investigate the interaction of Helicobacter pylori (Hp) and indomethacin in gastric mucosa injury of Balb/c mice. METHODS: Fifty specific pathogen free 6-week-old Balb/c mice were randomly divided into 5 groups: group A (n = 8, 25 mg/kg indomethacin was injected intraperitoneally every other day for 6 times, and 0.2 ml Hp culture containing 1 x 10(8) - 1 x 10(9) CFU/ml was inoculated intra-gastrically every other day for 6 times), B (n = 8, 1 x 10(8) - 1 x 10(9) CFU/ml Hp culture was inoculated intra-gastrically on the 1st, 2nd, and 3rd days, and since the 7th day 25 mg/kg indomethacin was injected intraperitoneally every other day for 6 tomes), group C (n = 9, 0.2 ml normal saline was inoculated intra-gastrically on the 1st, 2nd, and 3rd days and since the 7th day 25 mg/kg indomethacin was injected intraperitoneally every other day for 6 times), group D (n = 10, 1 x 10(8) - 1 x 10(9) CFU/ml Hp culture was inoculated intra-gastrically on the 1st, 2nd, and 3rd days, and since the 7th day 0.2 ml normal saline was injected intraperitoneally every other day for 6 times), and group E (n = 10, 0.2 ml normal saline was inoculated intra-gastrically on the 1st, 2nd, and 3rd days, then 0.2 ml normal saline was injected intraperitoneally since the 7th day every other day for 6 times). The mice were killed in the 4th week. The stomachs were observed morphologically. Tissues from gastric antrum underwent HE staining, silver staining, and urease examination. Rauws classification was used to grade the gastritis thus discovered. RESULTS: Two mice in group A, one mouse in group B and 2 mice in group C died. One mouse in group B was Hp silver staining negative. Twenty-six of the total 30 mice infected with Hp showed inflammation with an infection rate of 66.7%. Moderate to severe inflammatory reaction was seen in all 8 mice in group A with an average gastritis score of 6.38 and with Hp found in the epithelium. Two out of the 8 mice in group showed focal erosion. All mice in group B showed moderate inflammation and Hp in mucosa with an average gastritis score of 5.13. Focal erosion was seen in one mouse in group B. Light moderate inflammation was seen in all 9 mice in group C with an average gastritis score of 3.89. Mild inflammation was seen in all 10 mice in group D with an average gastritis score of 2.8. No erosion was found in groups C and D. No inflammation was seen in the mice in group E with an average gastritis score of 0.4. The differences between any 2 groups were all significant. CONCLUSION: Hp and indomethacin synergetically aggravate the gastric epithelial injury. Prior Hp infection may be a protective factor of indomethacin induced gastritis to a certain degree. PMID- 12899746 TI - [Helicobacter pylori inhibits cell growth and induces G1/S arrest in AGS gastric epithelial cells]. AB - OBJECTIVE: To observe the effects of Helicobacter pylori (H. pylori) on growth and cell cycle progression of AGS gastric epithelial cells. METHODS: AGS cells and H. pylori were co-cultured for 1 - 4 days. Vital cell numbers were counted by heamatocounter. Apoptosis was detected by fluorescence stain of cell nucleus and DNA agarose electrophoresis. Cell cycle was analyzed by flow cytometry technique. RESULTS: During co-culture of AGS cells and H. pylori, cell growth inhibition was observed. H. pylori induces apoptosis and cell cycle arrest from G1 to S phase accompanied by high expression of cyclinD1. CONCLUSION: Exposure of gastric epithelial cells to H. pylori causes block of G1 to S progression of cell cycle and alters expression of cyclinD1. The cell cycle arrest and the associated induction of apoptosis are responsible for the growth inhibition of gastric epithelial cells caused by H. pylori. PMID- 12899747 TI - [Cloning and immunogenicity of conservative region of adhesin gene of Helicobacter pylori]. AB - OBJECTIVE: To construct a candidate strain of Helicobacter pylori (Hp) that expresses the proteins of the conservative region of 4 adhesins (BabA2, AlpA, AlpB, and HopZ) and study its immunogenicity. METHODS: The DNA of Hp was extracted. Primers were designed according to the C-terminal structural gene sequence (called CB) of AlpA. The CB gene was amplified by PCR and inserted into the prokaryotic expression vector pET-22b (+) and expressed in BL21 (DE3) strain of Escherichia coli. The product of expression, CB, was purified by affinity chromatography and identified by Western blot analysis. ELISA assay was used to measure the CB-specific antibody in the specimens of serum of 55 Hp infected patients. Rapid urease test (RUT) was used on biopsy specimens collected by gastroscopy as parallel control. RESULTS: A recombinant plasmid pET-22b (+)/CB was constructed with the conservative region of the 4 adhesins. DNA sequencing showed one open reading frame of 588 bp encoding a polypeptide of 195 amino acids. The recombinant CB (rCB) protein, with a molecular weight of 22.5KD, amounted to 29% of the total bacterial protein. The purity of purified rCB was 96%. Western blot analysis showed that the rCB protein could be specifically recognized by the serum from Hp infected patients. The kappa coefficient was 0.76 for evaluation by ELISA and RUT results. CONCLUSION: CB has the potential to be used as a vaccine against Hp infection. PMID- 12899748 TI - [Treatment of hepatocellular carcinoma by transfecting interleukin-12 and interleukin-2 fusion gene intrasplenically, an experimental study]. AB - OBJECTIVE: To study the inhibitory effect of retroviral packaging cells injected intrasplenically encoding mouse interleukin-12 (mIL-12) and human interleukin-2 (hIL-2) fusion gene on the growth of hepatocellular carcinoma. METHODS: The retroviral vectors encoding mIL-12 gene, hIL-2 gene, and mIL-12 and hIL-2 genes, GCIL12EXPN, GCXEIL2PN, and GCIL12EIL2PN were constructed and then transfected into the retroviral packaging cells PA317 to construct cells PA317-GCIL12EXPN, PA317-GCXEIL2PN, and PA317-GCIL12EIL2PN. Rat hepatocellular carcinoma cells CBRH3 were implanted into the livers of Wistar rats to establish hepatoma animal model. Then the rats were divided into 5 groups to be injected intrasplenically with normal saline one day after the implantation (0.8 ml/rat, group I, n = 10), blank vector PA317-GCXEXPN one day after the implantation (10(7) cells/rat, group II, n = 10), PA317-GCIL12EXPN containing IL-2 gene (1 x 10(7) cells/rat 1, 3, 5, or 7 days after the implantation, group III, n = 40), PA317-GCXEIL2PN containing mIL 12 gene (1 x 10(7) cells/rat 1, 3, 5, or 7 days after the implantation, group IV, n = 40), and PA317-GCIL12EIL2PN containing IL-12-IL-2 fusion gene (1 x 10(7) cells/rat 1, 3, 5, or 7 days after the implantation, group V, n = 40) respectively. The rats surviving longer than 2 months were re-injected with hepatocellular carcinoma cells. The therapeutic effect, immune function and toxic effect were evaluated. CT was conducted on the liver before and after the experiment. Laparotomy was performed 3 and 7 days after treatment to resect some of the carcinoma tissues to undergo pathological examination and OX8 immunohistostaining. Serum mIL-12 and hIL-2 were detected one day before and 3, 7, 30, and 60 days after treatment. RESULTS: The average survival times of the rats treated with IL-12-IL-2 fusion gene at the first, third, fifth and seventh day after tumor implantation were 53.3 +/- 3.7 days, 49.3 +/- 4.2 days, 31.0 +/- 2.1 days, and 24.3 +/- 1.4 days respectively, longer than those treated with IL-2 gene (25.0 +/- 2.5 days, 23.5 +/- 2.0 days, 18.3 +/- 2.4 days, and 12.0 +/- 1.8 days respectively, P < 0.001), and those treated with IL-12 gene (39.0 +/- 4.8 days, 32.0 +/- 3.9 days, 23.0 +/- 2.5 days, and 19.4 +/- 2.1 days respectively, P < 0.001). Long survival (>or= 60 days) rate in the rats treated with IL-12-IL-2 fusion gene on the first and third day was 30%. The serum mIL-12 and hIL-2 levels in these rats remained high on the 60th day after treatment. The pathological study showed that the number of infiltrating lymphocytes in liver tumor tissues was increased in the IL-12-IL-2 fusion gene treatment group. CONCLUSION: The retroviral packaging cell line injected intrasplenically encoding mIL-12 and hIL 2 fusion gene inhibits the growth of hepatocellular carcinoma significantly in rats. The therapeutical efficacy of early administration is superior to that of late one. PMID- 12899749 TI - [Overexpression and genomic amplification of decoy receptor 3 in hepatocellular carcinoma and significance thereof]. AB - OBJECTIVE: To investigate the mRNA expression and genomic amplification of decoy receptor 3 (DcR3) in hepatocellular carcinoma (HCC) and the significance thereof. METHODS: RT-PCR was used to examine the expression of DcR3 mRNA in 48 cases of HCC tissues and the normal tissues adjacent to the tumor resected in operation and proved pathologically. The DcR3 gene amplification in the cancer tissues was examined by quantitative genomic PCR. The correlation between the DcR3 mRNA expression and genomic amplification of HCC was analyzed statistically. The differences in DcR3 expression and amplification in HCC with different clinical and pathological features were compared. RESULTS: DcR3 mRNA was detected in 29 out of the 48 HCC cases with a positive rate of 60.4%. No DcR3 mRNA expression was detected in the 48 cases of normal tissues adjacent to tumor. DcR3 mRNA was expressed significantly higher in HCC > 5 mm, at stages and, and with infiltration and metastasis than those 0.05). The DcR3 gene amplification in 48 cases of HCC ranges from 0.18 to 3.86-fold. Seven of the 48 cases of HCC showed significantly higher DcR3 gene amplification than the normal control tissues. The positive rate of DcR3 mRNA expression was 40% in the 25 HCC patients with the amplification fold < 1.0 (10/25), significantly lower than that in the 23 patients with the amplification fold > 1.0 (82.6%, 19/23), P < 0.01. The average DcR3 gene amplification fold was 1.53 +/- 0.82 in the patients with positive mRNA expression, significantly higher than that in those with negative expression (0.75 +/- 0.33, P < 0.01). The DcR3 gene amplification in HCC showed no relationship with the clinical and pathological feature of the patients (all P > 0.05). Sequencing analysis showed 3 point mutations in the DcR3 gene mRNA of HCC in one case of positive amplification product. CONCLUSION: DcR3 mRNA is overexpressed in HCC, but not in the normal tissues adjacent to the tumor. The expression of DcR3 gene is related to the size, clinical stage, infiltration, and metastasis of HCC. DcR3 gene amplification occurs in HCC and is correlated with the mRNA expression. Point mutation of the DcR3 gene exists in HCC. PMID- 12899750 TI - [Human papillomavirus infection and other risk factors for cervical cancer in Chinese and Australian patients]. AB - OBJECTIVE: To investigate the infection rate and genotypes of HPV in cervical cancer and analyse the clinicopathological characteristics and risk factors for cervical cancer with HPV infection in Chinese and Australian patients. METHODS: The human papillomavirus (HPV) DNA in cervical cancer specimens from 115 Chinese women and 79 Australian women was detected by polymerase chain reaction.HPV types were determined by sequencing and reverse hybridization. The relationship a between the HPV types and the clinicopathological characteristics of cervical cancer in Chinese and Australian women were analyzed. RESULTS: HPV DNA positive rate was 76. 5% in the Chinese women (88/115) and 89.9% in the Australian women (71/79, P = 0. 017). There were no significant differences of HPV 16 and HPV 18 prevalence between the two groups (P = 0. 806). HPV 58 and 59 were more prevalent in Chinese women (both 6. 8%) than in Australian women (1. 4% and 0). In China, HPV 58 prevalence rate was significantly higher in the Hunan region than in the Guangdong region (P = 0. 007). The mean ages at diagnosis was 44. 24 +/- 11. 31 years for Chinese and 53.08 +/- 16. 54 years for Australian women (P <0. 001). Combining the two groups, no relationship was found between HPV positivity/negativity and the FIGO stage, macroscopic features, histological type and grade of tumour, as well as pelvic lymphatic metastasis. However, HPV type wa significantly correlated with the histological type (P < 0. 001 ) and grade of cervical tumour (P = 0. 028). In comparison with the Chinese group, the Australian group presented more advanced cancers, a greater proportion of endocervical patterns, and more non-squamous cell carcinoma. There were no significant differences in grade of tumour and lymphatic node status between the two groups. CONCLUSION: The HPV infection rate in cervical cancer patients is significantly higher among Australians than among Chinese, which may be caused by more sexual transmitted diseases and more sex partners among the former group. HPV 58 and HPV 59 tie for third common genotypes in cervical cancers in China, however, these types are uncommon in Australia. There was a significant variation in types of HPV infection among different histological type and grade of cervical tumors. Both HPV 18 and 59 appear to be associated with the malignancy of cervical cancer. PMID- 12899751 TI - [The clinical value of three-dimensional computed tomographic angiography in embolization of acutely ruptured intracranial aneurysms with Guglielmi detachable coil]. AB - OBJECTIVE: To evaluate the clinical value of three-dimensional computed tomographic angiography (3D-CTA) in embolization of ruptured intracranial aneurysms with Guglielmi detachable coil (GDC). METHODS: From December 1998 to February 2002, 175 consecutive patients with acute subarachnoid hemorrhage (SAH) underwent 3D-CTA on the day of admission. All the patients with intracranial aneurysms diagnosed by 3D-CTA received emergency intraaneurysmal GDC embolization in case the ruptured aneurysm was suitable to endovascular therapy. RESULTS: Eighty patients with intracranial aneurysms received successfully emergency GDC embolization according to the results of 3D-CTA, all verified by digital subtraction angiography (DSA). 3D-CTA was superior to DSA in depiction of the 3D anatomy of aneurysms and parent arteries and provided valuable information for embolization of intracranial aneurysms prior to catheter angiography, especially for anterior communicating artery aneurysms. CONCLUSION: 3D-CTA discloses the existence of intracranial aneurysms early, establishes the indications of embolization prior to the catheter angiography, determines the side of insertion of guiding catheters, helps choose the appropriate coil size, and may become a routine diagnostic modality. PMID- 12899752 TI - [Treatment of acute ruptured aneurysm with Guglielmi detachable coil embolization]. AB - OBJECTIVE: To investigate the curative effect of Guglielmi detachable coil (GDC) on acute ruptured intracranial aneurysms. METHODS: The clinical data of 34 patients with acute rupture of intracranial aneurysms treated with early GDC embolization within 6 days after the rupture, 6 being of grade I, 23 of grade II, 4 of grade III and 1 of grade IV according to Hunt-Hess grading, were analyzed retrospectively. Fourteen patients were treated within 3 days, and 20 patients were treated in the 4th to 6th days after the rupture. Two cases of wide-neck aneurysm were embolized with Medtronic AVE stent supported GDC. Angiography was used to observe the effects. RESULTS: Angiography showed complete occlusion in 28 cases and 90% occlusion in 4 cases. One aneurysm was ruptured during the procedure. The caudal tip of GDC protruded to the parent vessel in 2 cases. Two cases failed to be embolized due to severe cerebrovascular spasm. There were no mortality and complications. No re-rupture occurred in the follow-up. CONCLUSION: Less invasive and with less risk and complication, early GDC embolization is one of the ideal therapeutic methods to manage acute ruptured aneurysms. PMID- 12899753 TI - [Endovascular treatment of intracranial aneurysms with mechanical detachable spiral-neurological]. AB - OBJECTIVE: To evaluate the efficacy of endovascular treatment of intracranial aneurysms with mechanical detachable spiral-neurological (MDS-N). METHODS: Thirty six patients with intracranial aneurysms were treated with MDS-N by Seldinger's approaches through femoral artery puncture. A catheter was sent into the aneurysm with proper-size MDS-N system and a spiral was pushed into the aneurysmal lumen. RESULTS: The aneurysms were completely occluded in 32 cases while partial occlusion was obtained in 4 patients. The parent arteries remained unobstructed. Follow-up of 3 - 6 months showed that all patients recovered well. No death or permanent serious complication occurred. CONCLUSION: Simple, safe, and low-cost, treatment of intracranial aneurysm with MDS-N is effective and suitable for use in larger hospital. PMID- 12899754 TI - [Application of the third generation cementing technique in total hip anthroplasty]. AB - OBJECTIVE: To investigate the effect of the third generation cementing technique in total hip anthroplasty (THA). METHODS: The clinical data of THA performed on 107 hips in 90 patients suffering from different kinds of orthopedic diseases, 54 males and 36 females, with an average age of 55 years and the average follow-up period of 36 months, were analyzed. RESULTS: According to the Harris scoring system, the 90 patients achieved a score of 34 (range 4 - 78) before the operation and achieved a score of 91 (range 64 - 100) after the operation. 97.1% of the patients showed good or excellent results during the short-term follow-up. CONCLUSION: The third generation cementing technique is rather effective in total hip anthroplasty. PMID- 12899755 TI - [Cytomegalovirus diseases after allogeneic hematopoietic stem cell transplantation (Allo-HSCT) and related risk factors]. AB - OBJECTIVE: To analyze the incidence of cytomegalovirus (CMV) diseases after allogeneic hematopoietic stem cell transplantation (allo-HSCT) and related risk factors. METHODS: The clinical data of 131 patients undergoing allo-HSCT August 1999 to July 2001 were analyzed retrospectively. RESULTS: Twenty-eight of the 131 patients developed CMV interstitial pneumonia (IP) with an incidence rate of 21.35%, 9 patients developed CMV enteritis with an incidence rate of 6.87%; in total 37 of the 131 patients developed CMV disease with the accumulative one-year incidence rate of 32.54%. Thirteen patients (35.14%)died of CMV diseases. Univariate analysis showed that accumulative incidence of CMV disease was associated with unrelated donor, II-IV degrees acute graft versus host disease (aGVHD), additional immunosuppressive therapy for GVHD, chronic graft versus host disease (cGVHD), antilymphocyte globulin (ALG)/monoclonal antibody (MoAb) administration, high dose of methylprednisolone (MP), plasma viramia-PCR (PV-PCR) positivity of patients, blood transfusion and pre-emptive treatment. Multivariate analysis showed that risk factors for CMV disease included plasma CMV-DNA positivity, immense amount of blood transfused, and additional immunosuppressive therapy for aGVHD, and pre-emptive therapy capable of turning PV-PCR positive reduced the incidence of CMV disease (RR: 3.309, 1.046, 2.242, and 0.346). CONCLUSION: CMV disease has a high morbidity and mortality in allo-HSCT patients. Patients with positive plasma CMV-DNA should receive pre-emptive therapy till their plasma CMV-DNA turns to be negative, especially in condition of severe aGVHD, immense amount of blood transfused, or additional immunosuppressive therapy. PMID- 12899756 TI - [Using GST-tag to capture protein interaction of an interferon-inducible systemic lupus associated gene, IFIT1]. AB - OBJECTIVE: To investigate the protein-to-protein interaction of interferon induced protein with tetratricopeptide repeats 1 (IFIT1), a newly discovered systemic lupus erythematosus (SLE) related up-regulated gene, and its possible function. METHODS: Peripheral blood of 40 SLE patients was obtained to extract total RNA and synthesized cDNA. Real-time PCR was used to determine the IFIT1 expression at transcript level. Peripheral blood of another 10 SLE patients was extracted to obtain specimens of white blood cell lysate. Molecular cloning and a modified gluthion S-transferase (GST)-pull down assay were used to capture the protein interacting with IFIT1 in the specimens of white blood cell lysate. MALDI TOF mass spectrometry (MS) was preformed to identify the captured protein that could interact with IFIT1. Twenty-nine sex and age-matched healthy persons were used as controls. RESULTS: By real-time PCR showed that the IFIT1Delta Ct value (x +/- s) was 2.344 +/- 1.200 in the SLE patients and was 3.734 +/- 1.274 in the controls (P < 0.001), showing a significant up-regulation in SLE patients. IFIT1 was cloned and GST-IFIT1 fusion protein was expressed in Escherichia coli. GST IFIT1 fusion protein was further purified using Glutathione Sepharose 4B column, and was treated as bait to capture prey from peripheral white blood cell lysate of SLE patients. MALDI-TOF MS detected protein interaction between Rho/Rac guanine nucleotide exchange factor and IFIT1. CONCLUSION: IFIT1 may interact with Rho/Rac guanine nucleotide exchange factor, and regulate the activation of Rho/Rac proteins, thus being involved in the pathogenesis of SLE. PMID- 12899757 TI - [A 3-amino acid deletion of apolipoprotein E found in 3 Chinese lipoprotein glomerulopathy patients]. AB - OBJECTIVE: To analyze the mutation of apolipoprotein E (apoE) gene so as to investigate the pathogenesis of lipoprotein glomerulopathy (LPG). METHODS: Restriction fragment length polymorphism methods was used to analyze the apoE genotypes of three Chinese patients with LPG and 3 of their relatives. Automated DNA sequencing was performed upon their apoE genes. RESULTS: A 9-bp deletion in exon 4 of apoE, resulting in a 3-amino acid deletion (residues 142-144-0) was identified. CONCLUSION: A 3-amino acid deletion of apoE has been discovered which is associated with LPG. PMID- 12899758 TI - [Bone marrow stromal cells of adult mice differentiate into smooth muscle cells in vitro]. AB - OBJECTIVE: To explore the potential of murine bone marrow stromal cells (MSCs) to differentiate into vascular smooth muscle cells so as to contribute to neointimal formation after vascular injury. METHODS: Murine MSCs were isolated from bone marrow and cultured in M199 with 10% FBS supplemented with medium conditioned from human smooth muscle cell culture to induce differentiation. Immunofluorescent technique was used to observe the morphology of cells to examine the smooth muscle characters. Vasoactive substance was added onto the cultured smooth muscle-like cells. Inverted microscopy was used to observe the contractile changes of the cells. RESULTS: After 6 passages of subculture in differentiation medium, MSCs demonstrated smooth muscle cell-like spindle-shaped morphology and assumed a hill-and-valley growth pattern. Immunofluorescent technique revealed positive signals for alpha-smooth muscle actin, calponin and smooth muscle myosin heavy chains. The smooth muscle cell-like cells displayed robust calcium transients. In response to stimulation of vasoactive substance the [Ca(2+)] of the cells instantaneously increased which was coupled by prominent cell contraction. CONCLUSION: MSCs can differentiate into vascular smooth muscle cells with contractile ability. This model may be useful in study of smooth muscle cell differentiation and the origin of neointimal cells after vascular injury. PMID- 12899759 TI - [Expression of MMP-9 and MMP-9 mRNA in gastric carcinoma and its correlation with angiogenesis]. AB - OBJECTIVE: To investigate the expression of MMP-9 and MMP-9 mRNA in the tissue and cell line of the gastric carcinoma and its correlation with invasion, metastasis and angiogenesis of the cancer. METHODS: Seventy-four specimens of resected human gastric carcinoma were collected. Human lowly differentiated gastric carcinoma cells of BGC823 line were cultured. Hematoxylin and eosin staining, immunohistochemistry, in situ hybridization, and zymography were used to examine the expression of MMP-9, MMP-9 mRNA, CD34, alpha-SMA, VEGFR-1, VEGFR 2. Invasion test was made to observe the number of cells penetrating the Matrigel. The relationship between the expression of MMP-9 and MMP-9 mRNA and the clinical parameters was analyzed. RESULTS: The positive rate of MMP-9 expression was 77.03% in gastric carcinoma tissues (57/74). Most of the MMP-9 positive cells were located at the invasive areas. The average microvascular density (MVD) of the 74 cases of gastric carcinoma was 73.76 +/- 11.50/200 x visual field. MVD was 75.46 +/- 10.98 in the MMP-9 positive group and was 68.08 +/- 11.71 in the MMP-9 negative group (P < 0.05). The positive rate of MMP-9 mRNA expression was 72.97% in the gastric carcinoma tissues (54/74). The expressions of MMP-9 and its mRNA were closely related to invasion, differentiation and lymph nodes status of the gastric carcinoma; MVD was closely related to the invasion and lymph nodes status of the gastric carcinoma. All BGC823 cells were MMP-9, MMP-9 mRNA, VEGFR-2 positive and VEGFR-1 negative. The number of invasive BGC823 cells was (18 +/- 4)/400 x in the MMP-9 antibody group and was (69 +/- 5)/400 x in the control group. BGC823 cells cultivated in culture dishes expressed the proenzyme of MMP-2 and MMP-9 stably. BGC823 cells cultivated on the surface of type I collagen expressed more MMP-2 proenzyme and active MMP-2. When BGC823 cells were incubated with VEGF, the content of MMP-9 proenzyme was increased by VEGF dose-dependently. CONCLUSION: Gastric cancer cells have the ability to produce and secret MMP-9 which can promote the invasion and metastasis of the neoplasm. PMID- 12899760 TI - [Differentially expressed genes in immunosuppressive and healthy mice after intranasal inoculation of Aspergillus fumigatus]. AB - OBJECTIVE: To screen out the differential expression genes between the immune suppressive host and healthy host after intranasal inoculation of Aspergillus fumigatus (A fumigatus). METHODS: Six immune suppressive mice and 6 healthy mice were inoculated with A, fumigatus intranasally. Three days after the inoculation the mice were killed and autopsied. The total RNA of the lungs was isolated. Gene chip technology was used to examine the differentially expressed genes. RESULTS: 66 differentially expressed genes were screened out of the 4098 genes. In the experimental group the genes encoding CD22, CD53, and 24p3 and the genes related to engulfment and digestion of macrophage or monocytes were up-regulated. The genes encoding surfactant associated protein C, immunoglobulin lambda chain and the genes related to pulmonary metabolism were down-regulated. The genes encoding IL18, CD8, and CD28 were not differentially expressed. CONCLUSION: The up regulation of CD22 gene may be one of the ways by which the host defense is inhibited. CD53 may take part in the regulation of engulfment of phagocytes. 24p3 protein, an acute stress protein, protects the host from excessive inflammatory response. PMID- 12899761 TI - [Evaluation of bone architecture and biomechanic properties by peripheral quantitative computed tomography in rats]. AB - OBJECTIVES: To evaluate the value of peripheral quantitative computed tomography (pQCT) in measuring bone architecture and biomechanic properties. METHODS: 50 virgin female Wistar rats six months old were randomly divided into 4 groups: (1) 8 rats were killed as baseline group; (2) 8 rats underwent sham operation and then were killed 14 weeks after (sham operation group); (3) 16 rats underwent bilateral ovariectomy (OVX) without further intervention. Six and 14 weeks after the operation each 8 rats were killed (OVX group); and (4) 18 rats underwent OVX too. After the OVX 9 of the 18 rats were treated with 17beta-estradiol 20 micro g/kg/d IH and 9 rats were treated with estradiol valerate 800 micro g/kg/d po for 8 weeks respectively. Then the 18 rats were killed (OVX plus estrogen group, O + E group). The right tibiae of the rats were taken for histomorphometric analysis, and the right femora were prepared for pQCT scanning and bone biomechanical measurement with indentation test and three-point bending test. RESULTS: Histomorphometric analysis showed that the trabecular volume of proximal tibia (Cn-BV/TV) in the OVX group was 8.1 +/- 1.4%, significantly lower than that in the sham operation group (19.5 +/- 1.5%, P < 0.01). pQCT scanning showed that the femoral trabecular bone mineral content (Trab BMC) in the OVX group was 1.7 +/- 0.3 mg/mm, significantly lower than that in the sham operation group (3.2 +/- 0.5 mg/mm, P < 0.01) and the femoral trabecular bone mineral density (Trab BMD) in the OVX group was 158 +/- 32 mg/mm(3), significantly lower than that in the sham operation group (320 +/- 39 mg/mm(3), P < 0.01). The cancellous maximal load (Can load) of the distal shaft of femur in the OVX group was 12.5 +/- 2.5 N, significantly lower than that in the sham operation group (45.9 +/- 3.2 N, P < 0.01). The cancellous stiffness (Can Stiff) of the distal shaft of femur in the OVX group was 226 +/- 48 N/mm, significantly lower than that in the sham operation group (396 +/- 72 N/mm, P < 0.01). The Can load of O + E group was 21.8 +/- 3.7 N, significantly higher than that in the OVX group (P < 0.05). The Can Stiff of the O + E group was 382 +/- 54 N/mm, significantly higher than that in the OVX group (P < 0.05). There were no significant differences in cortical bone determined by pQCT as well as biomechanic properties in measured by three point test after OVX and estrogen treatment. A significant positive correlation was shown between Trab BMD and Cn-TV/BV and between Trab BMD and Tb N (r = 0.88 and 0.73, both P < 0.01). Similarly, both Trab BMC and Trab BMD of the femur were significantly correlated with the Can load and Can Stiff determined by indentation test (r = 0.47 - 0.68, all P < 0.01). There was also a significant correlation of parameters measured by pQCT in cortical bone with the maximal load and stiffness for the femur midshaft, and the best correlation was found between the maximal load of femur midshaft and Crt BMC and Crt A (both r = 0.76 and P < 0.01). CONCLUSION: The geometric, densitometric and mechanical properties in cortical and trabecular bones of rat can be well described by pQCT. PMID- 12899762 TI - [Relationship between learning ability and memory and free radical in hippocampus of old rats]. AB - OBJECTIVE: To study the relationship between the amount of free radical in hippocampus and the learning ability and memory in old rats. METHODS: Morris water maze was used for 10 days to examine the learning scores, latency scores, and the loci of movement in the maze among 46 old rats (aged 20 months), 36 young rats (aged 6 months), and 34 adult rats (aged 12 months). Salicylate was injected intraperitoneally and hippocampus dialysis was performed. Chromatography was used to measure the basic values of 2,3DHBA and 2,5DHBA in dialysis fluid chemiluminometry was used to measure the superoxide dismutrase (SOD) in the sample. The rats were killed and their brains were taken out. Histological examination was conducted to calculate the number of neurons in CA1, CA4, and PM regions in hippocampus. RESULTS: The learning scores on the first, fifth, and tenth days were 36.5 +/- 5.9 sec, 38.6 +/- 5.9 sec, and 39.4 +/- 6.9 sec (P > 0.05) in the old rats; 60.2 +/- 5.4 sec, 156.8 +/- 5.8 sec, and 165.1 +/- 6.8 sec in the young rats; and 61.7 +/- 5.8 sec, 152.3 +/- 6.9 sec, and 168.7 +/- 6.5 sec in adult rats. The learning scores at any time point of old rats were significantly lower than those of the young and adults rats (all P < 0.01). The latency scores of the first, fifth, and tenth days were 25.7 +/- 1.2 sec, 27.5 +/ 1.9 sec, and 27.7 +/- 1.9 sec in the old rats without significant difference between any two of these values (P > 0.05), however, the 3 latency scores of the old rats were all significantly larger than those of the young and adult rats (21.8 +/- 1.7 sec, 5.9 +/- 1.0 sec, and 3.6 +/- 0.6 sec, and 21.8 +/- 1.6 sec, 5.0 +/- 0.9 sec, and 4.8 +/- 0.7 sec respectively, all P < 0.01). The basic value of 2,3DHBA after intraperitoneal injection of salicylate was 20.7 +/- 0.3 pmol/ml in the old rats, 5 times that of the young rats and 6 times that of the adult rats (P < 0.01). The basic value of 2,5DHBA after intraperitoneal injection of salicylate was 60.12 pmol/ml in the old rats, 6 times that of the young rats and 5.8 times that of the adult rats (P < 0.01). The SOD activity was 410 U x g(-1) x min(-1) +/- 50 U x g(-1) x min(-1), significantly lower than those in the young and adult rats (880 U x g(-1) x min(-1) +/- 62 U x g(-1) x min(-1) and 860 U x g( 1) x min(-1) +/- 60 U x g(-1) x min(-1) (both P < 0.01). The values of neuron density in CA1, CA4, and PM regions of the old rats were all significantly lower than those of the young and adult rats (all P < 0.01). CONCLUSION: The decrease of learning ability in old rat is positively correlated with the increase of free radicals and loss of neurons in hippocampus. PMID- 12899763 TI - [Management of critical severe acute respiratory syndrome and risk factors for death]. AB - OBJECTIVE: To investigate the management of critical severe acute respiratory syndrome (SARS) and possible risk factors for death. METHODS: Thirty-three patients with SARS referred to Guangzhou Institute of Respiratory Disease (GIRD) between December 2002 and March 2003 were studied retrospectively. Paired t-test using statistical software SPSS 10.0 was employed to compare the respiratory frequency and pulse O(2) saturation (SpO(2)) before and after noninvasive ventilation with continuous positive airway pressure (CPAP) on the 33 patients. Among them, 18 patients who presented with SpO(2) of 90% - 93% at rest under O(2) inhalation 5 L/min were further recorded for their SpO(2) during slight physical movement and CPAP. The possible death-related risk factors including age, underlying diseases, leucocytosis, thrombocytopenia, and lymphopenia were analyzed by the Mantel-Haenszel chi(2) test. RESULTS: The respiratory frequencies were significantly decreased and SpO(2) was improved in the 33 patients after one hour of noninvasive ventilation. The SpO(2) in the latter 18 patients fell during slight physical movement and improved one hour after CPAP (p < 0.01). The RRs of the five death predictors concerning thrombocytopenia, age (> 50 yrs, underlying diseases, leucocytosis, and lymphopenia were 25.83, 8.57, 6.40, 1.64, and 1.17, respectively, with the 95% CI of 3.64 - 183.59, 1.94 - 37.83, 1.75 - 23.33, 0.38 7.05 and 0.16 - 8.48, respectively. Management with corticosteroids effectively ameliorated and suppressed the development of pulmonary fibrosis. CONCLUSIONS: Noninvasive ventilation relieves dyspnea and SpO(2) in patients with critical SARS, and should also be employed in those with SpO(2) of 90% - 93% at rest under O(2) inhalation 5 L/min. Noninvasive as an add-on management may probably cut down on the dosage and duration of corticosteriod therapy. Among 5 possible risk factors, 3 were recognized as death-related, turning out to be thrombocytopenia, age (> 50 yrs) and underlying diseases. PMID- 12899764 TI - [Severe acute respiratory syndrome in 78 patients: a retrospective study]. AB - OBJECTIVE: To analyze the clinical features of severe acute respiratory syndrome (SARS) and the diagnosis and treatment of the disease. METHODS: Seventy-eight patients with SARS referred to the Guangzhou Institute of Respiratory Diseases (GIRD), China from December 22, 2002 to March 2003 were studied retrospectively. The data reviewed included those of clinical manifestations, laboratory investigation and roentgenology. RESULTS: The patients consisted of 42 men and 36 women, aged 20 - 75 years (mean 37.5 +/- 11.6 years), including 44 infected health-care professionals. Clinical symptoms of these patients were fever (100%), coughing (88%), and dyspnea (80%). Routine blood test revealed WBCs <4.0 x 10(9)/L in 12 patients (15%), (4.0 -10.0) x 10(9)/L in 49 (63%), and over 10.0 x 10(9)/L in 17 (22%) [average (7.6 +/- 5.0) x 10(9)/L]. The level of neutrophilic granulocyte was 0.75 +/- 0.13 and that of lymphocyte was 0.18 +/- 0.11. Chest X ray and CT scanning revealed changes related to pneumonia. The transmission of the disease was likely via close contact with contagious droplets. The prevalences of acute lung injury (ALI,37 patients) and acute respiratory distress syndrome (ARDS, 21 of the 37 patients) were considerably high among the patients. Seven patients who developed ARDS complicated with MODS died. CONCLUSIONS: A history of close contact, fever, X-ray signs of pneumonia and normal or lowered WBC counts are favorable for the diagnosis of SARS. Recognition of ALI as the important index for critical SARS and comprehensive supportive management are of paramount importance in decreasing the mortality of patients with SARS. PMID- 12899765 TI - [Serum antibodies detection for serological diagnosis of severe acute respiratory syndrome]. AB - OBJECTIVE: To evaluate the significance of serum specific antibodies against severe acute respiratory syndrome (SARS) virus in the diagnosis of SARS and the concordance of IFA and ELISA in the antibodies measurements. METHODS: Specific antibodies of IgM and IgG against SARS virus in 267 serum specimens from patients with SARS and 132 from non-SARS patients or healthy persons were assayed with IFA or ELISA. The sensitivity, specificity, positive predictive value (ppv), negative predictive value (NPV), and accuracy were used to evaluate the significance of the antibodies in diagnosis of SARS. The concordance of different measurements was tested with Kappa. RESULTS: The positive rate of specific IgM and IgG detected by IFA increased sharply on the eleventh day of the disease. The sensitivity, specificity, PPV, NPV, and accuracy of IgM in diagnosis of SARS on eleventh day or later were 65.6%, 100.0%, 100.0%, 71.0% and 81.3%, respectively, and IgG were 91.1%, 97.0% 97.3%, 90.1%, and 93.8%, respectively. Similar results could be seen when IgM and IgG were detected by ELISA. The Kappa of IFA and ELISA for IgM and IgG were 0.640 and 0.779 respectively. CONCLUSIONS: The antibodies against SARS virus can be detected in the mid-and late stage of SARS. The antibodies can be used in the serological diagnosis of SARS. There is high concordance of IFA and ELISA results in measurement of SARS antibodies. PMID- 12899766 TI - [Measurement of subsets of blood T lymphocyte in 93 patients with severe acute respiratory syndrome and its clinical significance]. AB - OBJECTIVE: To investigate the changes of subsets of blood T lymphocyte in patients with severe acute respiratory syndrome (SARS) and their clinical significance. METHODS: Subsets of blood T lymphocyte in 93 patients with SARS were detected by flow cytometer. The patients comprised 40 men and 53 women, aged 17 - 88 years (average 44 years). The results detected in 64 normal subjects and 50 patients with AIDS served as controls. RESULTS: The numbers of CD(3)(+), CD(4)(+), and CD(8)(+) lymphocytes all significantly decreased in acute phase of SARS patients compared with those in normal persons. Their findings was different from what we observed in patients with AIDS who had decreased CD(4)(+), but increased CD(8)(+) counts. The counts of CD(3)(+), CD(4)(+), and CD(8)(+) lymphocytes decreased more apparently in patients with critical SARS. All the five patients who died had CD(4)(+) counts less than 200/microl. As the patients' conditions improved, the counts of CD(3)(+), CD(4)(+), and CD(8)(+) gradually returned to normal. CONCLUSION: The patients with SARS were found having damage of cellular immunity markedly. PMID- 12899767 TI - [Roentgenography and CT appearance in patients with severe acute respiratory syndrome]. AB - OBJECTIVE: To evaluate the appearance and dynamic change of chest roentgenography and CT in patients with severe acute respiratory syndrome (SARS). METHODS: Serial roentgenography and CT of 61 confirmed SARS patients were analyzed. RESULTS: The onset symptom was fever in all patients. In the early phase of SARS, the main abnormal appearance was single or bilateral localized patchy clouding opacity on roentgenography, which progressed rapidly. The area of opacity peaked at about 2 weeks, involving the majority area of the lung in 57% of the patients. The area involved was correlated with the severity of clinical symptoms. The opacity disappeared gradually in 5 weeks in 84% of the patients. Lungs complicated infection were found in 6 patients with a prolonged duration of the illness. Among the 61 patients, 4 died. CT scan was performed in 16 SARS patients in the recovery phase, of whom 13 showed lung fibrosis. CONCLUSIONS: Lung infiltration in early phase develops rapidly and involves most area of the lung in advanced phase in most SARS patients. Lungs fibrosis was found in some patients in the recovery phase. PMID- 12899768 TI - [Different cytokine profiles in usual interstitial pneumonia and nonspecific interstitial pneumonia]. AB - OBJECTIVE: To study the distribution, the expression and the significance of TGF beta(1), b-FGF, IL-8, IL-13 and IFN-gamma in different lung tissue compartments in usual interstitial pneumonia/idiopathic pulmonary fibrosis (UIP/IPF) and nonspecific interstitial pneumonia (NSIP). METHODS: Specimens were obtained by open or video-assisted thoracoscopic lung biopsy from patients with UIP (n = 5) and NSIP (n = 8). Control specimens were obtained by surgical lobectomy from patients with primary lung cancer (n = 5). The distribution of these cytokines in lung tissues was observed by semi-quantitative method using immunohistochemical staining. RESULTS: TGF-beta(1), IL-8 and b-FGF were localized in alveolar epithelial cells, alveolar macrophages, and the bronchial epithelium. Overall intensity of TGF-beta(1), IL-8 and b-FGF expression in UIP was stronger in comparison with NSIP. IL-13 was distributed in alveolar epithelial cells, alveolar macrophages and interstitial mononuclear cells. Its expression in UIP was similar to that in NSIP. IFN-gamma was expressed mainly in interstitial mononuclear cells. Its expression in NSIP was stronger than that in UIP. The ratio of IL-13 to IFN-gamma in UIP (2.18 +/- 0.76) was significantly higher than that in NSIP (0.95 +/- 0.28) or that in the control (0.91 +/- 0.16) (P < 0.05, UIP versus NSIP or control), whereas the ratio of IL-13 to IFN-gamma in NSIP was similar to that in the control. In normal lungs, only alveolar macrophages expressed these cytokines. CONCLUSION: The different expression of TGF-beta(1), IL-8 and b-FGF in UIP and NSIP and the balance of IL-13/IFN-gamma may be involved in the different pathogenesis in these two diseases. PMID- 12899769 TI - [The effect of a mouse IL-12 plasmid on airway inflammation and cytokine production in a mouse asthmatic model]. AB - OBJECTIVE: To investigate the effect of a mouse IL-12 gene expressive plasmid (mIL-12 plasmid) on the airway inflammation and the cytokine production in asthmatic mice and to study the possible mechanisms. METHODS: A mouse model of asthma was established by sensitization with ovalbumin (OVA). Forty-one BALB/c mice were divided into six groups including an asthmatic model group (group A, eight mice, sensitized with OVA plus challenging with OVA by aerosol), a model control group (group B, six mice, sensitized with OVA plus aerosolizing with normal saline), a mIL-12 plasmid prevention group (group C, eight mice, receiving intramuscularly mIL-12 plasmid 100 micro g on day 1, day 3, and day 5), a mIL-12 plasmid treatment group (group D, eight mice, receiving intramuscularly mIL-12 plasmid 100 micro g on day 14, day 16, and day 18), an empty plasmid prevention group (group E, five mice, receiving intramuscularly empty plasmid 100 micro g on day 1, day 3, and day 5), and an empty plasmid treatment group (group F, six mice, receiving intramuscularly empty plasmid 100 micro g on day 14, day 16 and day 18). The number of EOS and the concentration of IL-4, IL-5 and IFN-gamma in the mouse bronchoalveolar lavage fluids (BALF) were detected. RESULTS: The number of EOS and the concentration of IL-4, IL-5 and IFN-gamma from group B were (0.01 +/- 0.03) x 10(8)/L, (24 +/- 4) pg/ml, (33 +/- 6) pg/ml, (725 +/- 59) pg/ml,respectively; those from group C were (0.06 +/- 0.04) x 10(8)/L, (43 +/- 13) pg/ml, (63 +/- 10) pg/ml, (626 +/- 60) pg/ml, respectively, and those from group D were (0.11 +/- 0.12) x 10(8)/L, (38 +/- 14) pg/ml, (66 +/- 14) pg/ml, (661 +/- 40) pg/ml, respectively; the difference was significant as compared with those from group A [(2.97 +/- 1.20) x 10(8)/L, (122 +/- 45) pg/ml, (126 +/- 34) pg/ml, and (435 +/- 49) pg/ml] (P < 0.001). The number of EOS and the concentration of IL-4, IL-5 and IFN-gamma from group C and group D also showed significant difference in comparison with those from group E [(1.96 +/- 0.93) x 10(8)/L, (110 +/- 24) pg/ml, (112 +/- 11) pg/ml and (464 +/- 51) pg/ml], and group F [(2.11 +/- 0.90) x 10(8)/L, (88 +/- 17) pg/ml, (107 +/- 6) pg/ml and (481 +/- 64) pg/ml] (P < 0.01). CONCLUSION: The mIL-12 plasmid can significantly inhibit airway inflammation. Its regulatory effect on the balancing of Th1/Th2 cytokines may be a possible mechanism. PMID- 12899770 TI - [The inhibitory mechanism of nitri-oxide synthase gene transfection on hypoxia induced proliferation of rat pulmonary arterial smooth muscle cells]. AB - OBJECTIVE: The underlying mechanism by which nitric-oxide synthase (iNOS) gene transfer inhibits hypoxia-induced PASMCs proliferation remains unknown. The aim of this study is to investigate if iNOS gene transfer to PASMCs during hypoxia has any effect on cell cycle progression. METHODS: Using the cationic liposome mediation method, we transfected a recombinant pLNCX/Inos vector into rat PASMCs. The instantaneous transgenic expression and the function of the recombinant protein were detected. Cell cycle analysis was performed by flow cytometry and cell proliferation assay by [(3)H] thymidine incorporation. The proteins involved in cell cycle control (P27 and P21) were determined by RT-PCR and flow cytometry. RESULTS: iNOS expression was detected in the transfected PASMCs. NO(2)(-) levels were increased in iNOS-transfected cells as compared to the untransfected cells. Expression of iNOS in rat PASMCs under hypoxia resulted in a delay in inhibition of DNA synthesis and cell cycle progression. The incorporation of [(3)H] thymidine in iNOS-transfected group (15,145 +/- 1,514) dpm was significantly lower than those in the hypoxia group (18,011 +/- 2,521) dpm (P < 0.01). The G(0)/G(1) cell cycle arrest rate in the iNOS-transfected group (67.8%) was significantly higher than those in the hypoxia group (46.8%) (P < 0.01). The protein level of P27 was down-regulated by hypoxia but not in iNOS-transfected cells under hypoxia, and the level of the latter was similar to that under normoxia. CONCLUSIONS: Pre-transfer of iNOS gene to PASMCs under hypoxia inhibits cell proliferation via blocking P27 down-regulation, which is an important mechanism for the delay of cell cycle progression. PMID- 12899771 TI - [Clinical feature and airway inflammation in eosinophilic bronchitis]. AB - OBJECTIVE: To study the clinical features and airway inflammation in eosinophilic bronchitis (EB) and the treatment outcomes. METHODS: Irwin's anatomic protocol for diagnosing chronic cough was used in 86 patients with chronic cough, and induced sputum by hypertonic saline aerosol inhalation was performed. Differential cell counts were performed in induced sputum, and eosinophilic cationic protein (ECP) was measured with fluoroimmunoassay, while interleukin-8 (IL-8) was measured with enzyme-linked absorbed immunoassay. EB was diagnosed according to Gibson's criteria and treated with inhaled budesonide 200 - 400 micro g twice daily for four weeks, and in some patients oral prednisone 10 - 15 mg/d or methyl-prednisone 8 - 12 mg/d was given for one week. RESULTS: 13 (15%) out of 86 patients with chronic cough were diagnosed as having EB. Dry cough was the major compliant and all had normal lung function with negative histamine provocation test. The Eos count was 0.1862 +/- 0.1632 and the concentration of ECP (2.53 +/- 2.07) mg/L in induced sputum were significant higher in patients with EB as compared with those normal subjects (P < 0.01). The cough disappeared in all patients at the end of one week of inhaled or orally administered corticosteroids. CONCLUSION: EB, an eosinophilic airway inflammation, is one of important causes of chronic cough and responds well to corticosteroid therapy. PMID- 12899772 TI - [Dynamic changes of T-lymphocytes and immunoglobulins in patients with severe acute respiratory syndrome]. AB - OBJECTIVE: To study the immunological changes in patients suffered from severe acute respiratory syndrome (SARS). METHODS: T-lymphocyte subsets of peripheral blood from 124 patients with different phases of SARS were measured by flow cytometry. Immunoglobulins were examined simultaneously. Blood specimens from 51 patients with AIDS and 57 normal persons were collected as controls and subjected to the same measurements. RESULTS: The levels of CD3(+), CD4(+) and CD8(+) T lymphocytes in SARS group [(589 +/- 435) x 10(6)/L, (316 +/- 267) x 10(6)/L and (215 +/- 165) x 10(6)/L, respectively] were significantly lower than those of the normal group (P < 0.01). When compared the SARS group with AIDS group, we found the level of CD4(+) T-lymphocyte in the former was higher (P < 0.01), while the CD3(+) and CD8(+) T-lymphocytes were lower (P < 0.01). There were 54 (44%) patients in SARS group and 39 (76%) patients in AIDS group showed low levels of CD4(+) T-lymphocyte (< 200 x 10(6)/L). The count of T-lymphocyte subsets of SARS group decreased to the lowest level during the 10(th) and 12(th) day after the onset of symptoms. Both serum IgG and IgE in SARS group [(15.1 +/- 8.5) g/L and (0.6 +/- 0.9) g/L, respectively] were higher than those of the normal group. CONCLUSION: The levels of CD3(+), CD4(+) and CD8(+) T-lymphocyte subsets in peripheral blood of SARS patients decrease significantly and go down to the lowest point at days 10 approximately 12 after onset of the disease. PMID- 12899773 TI - [Epidemiologic features, clinical diagnosis and therapy of first cluster of patients with severe acute respiratory syndrome in Beijing area]. AB - OBJECTIVE: Investigate the features of outbreak epidemic, clinical disease progression of the first SARS cases in Beijing and evaluate the efficacy of therapeutic regimen. METHODS: Twenty-nine patients (11 men and 18 women, 20 - 74 years old age range) were diagnosed with infectious SARS and admitted in our hospital from the March 5th to April 14th, 2003 in this study. The data of clinical presentation and disease progression of all the patients including index subject as the infectious SARS resource patient, her family infected members and 21 health care workers were abstracted. RESULTS: The first SARA outbreak in Beijing was characterized with the cluster feature of resource patient family members and health care providers. The incubation period ranged from 2 to 14 days. All the patients had a fever (temperature > 38 degrees C for over 24 hours) and other manifestations as reported before. Serial chest radiographs showed progressive pathologic air-space disease. Twenty patients showed the severe syndrome with various time ranged from 1 day to 14 days. Two patients died of progressive acute respiratory distress disease. The histologic analysis of one death patient showed diffuse alveolar damage in the two lungs. Twenty-six patients receiving the combined therapy including use of corticosteroid, antiviral ribavirin agents after the onset of symptoms and showed they had an acute self-limited disease course. The oldest patient (74 year old, male) received the healthy convalescent plasma infusion (50 ml) from recovered SARS subject and completely recovered within 21 days, having a shorter disease course. CONCLUSION: SARS is a kind of new self-limited and acute infectious disease. Early diagnosis, early isolation, early antiviral therapy for patients and efficient prevention for health care providers are urgently recommended. In particular, a combinational therapy of use of antiviral agents, preventive antibacterial antibiotics and pulsed dosage of corticosteroid can efficiently raise the clinical recovery rate and decrease mortality of SARS patients. PMID- 12899774 TI - [Surveillance on drug resistance of gram-negative bacilli isolated from hospital acquired infections and community acquired infections (2000 - 2001)]. AB - OBJECTIVE: To determine the resistance rates of gram-negative bacilli isolated from patients with hospital acquired infections (HAI) or community acquired infections (CAI) in the 34 study wards of 13 hospitals located at different areas in China. METHODS: 1596 strains of gram-negative bacilli from the 2401 pathogenic strains isolated from 13 hospitals 1 July 2000 to 30 June 2001 were collected for susceptibility test using international standard two-fold agar dilution method. According to the criteria of guidelines of NCCLS (2001), MIC(50) and MIC(90) were detected for antibacterial activity of antimicrobial agents and R%, I%, and S%, were calculated to show the resistant, intermediate and susceptible rates of bacteria to the compounds tested. RESULTS: 15 species of gram-negative pathogens were isolated and most of them could be isolated from both CAI and HAI patients. The resistance rates of isolates, especially Enterobacter cloacae, Serratia spp. and Acinetobacter spp. from HAI were higher than those of CAI. The carbopenems were the most active compounds tested against Enterobacteriaceae. Cefoperozone/sulbactam and cefprime demonstrated excellent activity against most of Gram-negative bacilli. Most of the Gram-negative bacilli were still rather susceptible to cefatazidime, a third generation cephalosporin, with a susceptibility rate of 70% to 100%. Fluoroquinolones showed strong and broad spectrum activity against Gram-negative bacilli, but more than 50% of Escherichia coli strains were resistant to fluoroquinolones, and the susceptible rates to fluoroquinolones of Pseudomonas aeruginosa and Haemophilus influenzae had decreased. CONCLUSION: (1) The results obtained in the two surveillance-years, 2000 - 2001 and 1998 - 1999, were similar in the bacterial resistance pattern and the trend of rising of resistant rates for some Gram-negative pathogens. (2) The resistance rates of isolates from HAI patients were higher than those of CAI. PMID- 12899775 TI - [Antimicrobial resistance of Escherichia coli isolates collected from inpatients and outpatients]. AB - OBJECTIVE: To investigate the antimicrobial resistance of Escherichia coli (E. coli) isolates collected from the inpatients in the departments of medicine, surgery, and pediatrics, and intensive care unit (ICU), and from the outpatients. METHODS: Disc diffusion test was used to study the antimicrobial resistance of 3909 strains of E. coli collected from the inpatients in the departments of medicine, surgery, and pediatrics, and Intensive care unit (ICU), and from the outpatients, mostly isolated from urine, sputum, blood, and different secreta in the year 2001. WHONET 5 software was used for analysis of the antimicrobial resistance; and significant differences were tested by chi(2) to compare the resistance rates to antibiotics. RESULTS: The incidences of extended-spectrum beta-lactamases producing strains were 11.2% (195/1737), 14.3% (141/983), 17.7% (28/158), 19.7% (24/122) and 8.4% (76/909) in the strains of E. coli isolated from the inpatients in the departments of medicine, surgery, and pediatrics, and ICU and from the outpatients respectively, with a detectable rate among the outpatients significantly lower than that among the inpatients (P < 0.005), and a detectable rate among the inpatients in the department of medicine significantly lower than those among the inpatients in the department of pediatrics and ICU (both P < 0.05). The resistance rates to cefazolin, cefotaxime, gentamicin and aztreonam of the isolates from the outpatients were significantly lower than those of the inpatients (all P < 0.05). The resistance rates to amoxicillin/clavulanic acid, ceftazidime, cefepime and amikacin of the isolates from the outpatients were significantly lower than those of the inpatients in the department of surgery and ICU (all P < 0.05); The resistance rates to ciprofloxacin and trimethoprim/sulfamethoxazole of the isolates from the outpatients were significantly lower than those from the ICU patients (both P < 0.05). The resistance rates to cefazolin and cefotaxime of E. coli isolates collected from the inpatients in the department of medicine were significantly lower than those of the isolates from the department of surgery and ICU (all P < 0.01); the resistance rates to gentamicin of the isolates from the department was significantly lower than that of the isolates from the department of surgery (P < 0.05). The resistance rates to amoxicillin/clavulanic acid and aztreonam of the isolates from the department of medicine were significantly lower than those of the isolates from the ICU (both P < 0.05). The resistance rate to ciprofloxacin of isolates from the inpatients in the department of pediatrics was significantly lower than that of the other isolates (all P < 0.01). CONCLUSIONS: It is of guiding significance for empirical use of antimicrobial agents in clinic to study on the resistant rates of the strains of E. coli isolated from different departments in hospital. PMID- 12899776 TI - [Analysis of antimicrobial resistance of clinical isolates of enterococci from Beijing and other areas in China]. AB - OBJECTIVE: To analyze the antimicrobial resistance of enterococci in Beijing and other areas in China. METHODS: Disk diffusion method was used to test the antimicrobial resistance of 1978 enterococcal isolates from 4 teaching hospitals in Beijing and 53 grade 3 A hospitals in other areas of China The laboratory data were analyzed by WHONET5 statistic software. RESULTS: Enterococcus. faecalis isolates and E. facium isolates accounted for 80.7% and 17.1% of the enterococcus isolates in Beijing and 79.4% and 15.2% in other areas respectively. The enterococcus isolates from urine, sputum, secretion with pus, blood, and bile accounted for 33.7%, 22.9%, 9.8%, 4.4%, and 3.0% of the enterococcus isolates in Beijing and 28.4%, 16.3%, 14.6%, 5.0%, and 4.5% in other areas respectively. The resistance rates of E. faecalis to vancomycin, teicoplanin, ampicillin, and high level gentamicin were 2.0%, 1.4%, 17.7%, and 47.4% in Beijing, and 5.1%, 3.7%, 24%, and 49.2% in other areas; The resistance rates of E. faecium to vancomycin, teicoplanin, ampicillin, and high -level gentamicin were 1.5%, 2.6%, 64.2%, and 64.9% in Beijing, and 2.1%, 3.1%, 68.5%, and 62.3% in other areas respectively. The vancomycin-resistant rate of the total isolates of enterococci was 3.2%. CONCLUSION: Enterococcus faecalis is the main species in the enterococcus isolates from clinical samples. E. facium, as compared with E. faecalis resists many antimicrobial agents more strongly. Both of E. faecalis and E. facium show the least resistance to vancomycin and teicoplanin. PMID- 12899777 TI - [Changes of clinical aspect of primary liver cancer in China during the past 30 years--control study for 3,250 cases with primary liver cancer]. AB - OBJECTIVE: To study the changes of the clinical aspects of primary liver cancer (PLC) during the past 30 years. METHODS: The clinical data of 3,250 patients with PLC, from 10 regions of China were collected, analyzed, and compared with the clinical data of 3254 PLC cases 30 years before. RESULTS: The 3,250 patients were aged 1- 80, with an average age of 49.1 years, younger than those 30 years before (43.7 years). The male to female ratio was 2.3:1, lower than that 30 years before (7.7:1). 73.5% of them sought medical advice within 3 months after the onset in comparison of 63.8% 30 years before. Compared with those 30 years before the symptoms and signs were alleviated in general. The HBsAg positive rate was 81.0%, the HCV-Ag positive rate was 13.2%, and the alpha-fetoprotein positive rate was 75%. 1912 cases underwent pathological examination of which 91.8% were diagnosed as with hepatocellular carcinoma. The overall resection rate was 46,3%. Those of early, median, and late stages accounted for 29.9%, 51.5%, and 18.6% respectively in comparison with the rates of 0.4%, 47.0%, and 52.6% 30 years before. The one year survival rate, three-year survival rate, and five-year survival rate were 66.1%, 39.7%, and 32.5% respectively for the whole group, 93.5%, 70.1%, and 59.1% for the early stage patients, and 65.3%, 30.5%, and 23.5% respectively for the median stage patients. The half-year survival rate and one-year survival rate of the late stage patients were 52.5% and 14.7% respectively. Compared with the data 30 years before a lower percentages of the patients died of hepatic coma, hemorrhage of upper digestive tract and hemorrhage due to rupture of tumor, and a higher percentage of then died of asthenia universalis and other causes. CONCLUSION: In comparison with the situation 30 years ago, PLC can be diagnosed earlier. More patients undergo resection. The prognosis of PLC has been improved greatly. PMID- 12899778 TI - [Role of leptin in the pathogenesis of obesity-related hypertension]. AB - OBJECTIVE: To assess the role of leptin in the pathogenesis of obesity-related hypertension and the relationship between blood pressure (BP), and body mass index (BMI), insulin resistance and leptin were examined. METHODS: 560 non diabetic men, aged 35 - 75, selected from volunteers of health screening test during 2 000 were divided into hypertension group (n = 321, BP >or= 140/90 mm Hg without antihypertensive medication). and normal blood pressure group (n = 239). The body weight, waist hip ratio,BP, plasma glucose, serum lipids, true insulin (TI) and leptin were measured after overnight fast. Insulin sensitivity was assessed by the HOMA insulin resistance index (HOMA-R). RESULTS: Fasting leptin level showed good correlation with BMI, fasting TI, HOMA-R, BP and also triglycerides (all P < 0.01). After adjustment for age, BMI and HOMA-R, serum leptin was still positively correlated to SBP (r = 0.11, P < 0.05), and was significantly higher in hypertensive subjects than in normotensive subjects (geometric mean 6.4 vs 4.7 micro g/L, P < 0.001). Logistic regression analysis demonstrated that leptin remained significantly associated with hypertension after adjustment for potentially confounding factors. CONCLUSION: Leptin may play an important role in the pathogenesis of obesity related hypertension. PMID- 12899779 TI - [Cervical posterior laminoplasty operation using T-saw and coralline hydroxyapatite spacer]. AB - OBJECTIVE: To assess the efficacy of using T-saw and coralline hydroxyapatite (CHA) spacer in the cervical spinous process-splitting laminoplasty. METHODS: 38 patients were operated with cervical laminoplasty using T-saw and CHA spacer, among them there were 31 multisegmental cervical spondylotic myelopathy patients, 5 ossification of posterior longitudinal ligament (OPLL) patients, 2 anterior fusion failure patients. All the patients were evaluated by JOA scores. RESULTS: we followed the 38 patients for average 26 months. Numbness,strength and gait were improved obviously. The flat rate of spinal cord in MRI improved 0.23;CHA fusion percentage 74%; no primary spinal cord injury was found. Average operation time was 2 hour 50 min;average bleeding during the operation was 280 ml; The average JOA score improve rate was 61%. CONCLUSION: The methods, using T-saw and CHA spacer in the cervical spinous process-splitting laminoplasty, which was characterized by simple, short time on operation and thorough decompression of spinal cord, was a ideal optional method in the cervical laminoplasty operation. PMID- 12899780 TI - [Surgical treatment of degenerative scoliosis]. AB - OBJECTIVE: To investigate the techniques and indications of treatment of degenerative scoliosis. METHODS: The clinical data of 45 degenerative scoliosis patients, 18 males and 27 females, aged 41 - 77, all suffering from severe low back pain, were analyzed. Twenty of them underwent posterior lumbar interbody fusion with pedicle screws combined with correction of scoliosis. Sixteen patients underwent depression of vertebral canal and pure internal fixation and fusion of pedicle. Eight patients underwent pure depression, such as laminectomy or incision of intervertebral foramen. The corrective effects of scoliosis, condition of intervertebral fusion, and complications were observed. RESULTS: All operations were performed successfully and all the patients made a remarkable recovery. During the 13 months' follow-up, 88% of the patients showed remarkable relief from pain and claudication and improved life quality. No loss in correction angle and height of disc space was found among the patients who had undergone internal fixation and interbody fusion. The residual symptom of low back pain was more manifest in the patients who had not undergone internal fixation than in those who had. CONCLUSION: Posterior lumbar interbody fusion with pedicle screws is safe and effective in the treatment of severe degenerative scoliosis. PMID- 12899781 TI - [Expression of hyaluronic acid and its receptor in the process of wound healing in different skin tissues and its significance]. AB - OBJECTIVE: To explore the expression of hyaluronic acid (HA) and its receptor, Cluster of differentiation 44 (CD44) in proliferative scar and in the process of wound healing of normal human adult skin and fetal skin, and the effect of HA and its receptor on the process of human fetal skin scarless healing. METHODS: An incision and then a hypodermic cavity were made on each side of the dorsal median line of 32 female adult BALB/c rats. Skin grafts from 8 human fetuses delivered by natural abortion, full-thickness skin grafts from 8 normal adults undergoing plastic operation, skin wound sample from the donor sites in legs of 8 patients undergoing dermatoplasty with intermediate split thickness skin graft, and proliferative scar from 8 patients of plastic surgery, non-adult and adult, were grafted into the hypodermic cavities. The levels of HA and its receptor were examined by radioimmunoassay, immunohistochemistry and flow cytometry. RESULTS: The level of HA in normal fetal skin was 143 micro g/g +/- 10 micro g/g, 283 micro g/g +/- 12 micro g/g 12 hours after injury, 315 micro g/g +/- 12 micro g/g one days after injury, reached the peak (321 micro g/g +/- 12 micro g/g) 3 days after injury, and then decrease, became 319 micro g/g +/- 11 micro g/g one week after injury (P > 0.05 in comparison with that 3 days after injury). The level of HA in normal fetal skin was 143 micro g/g +/- 10 micro g/g, significantly higher than that in normal adult skin (51 micro g/g +/- 4 micro g/g), skin wound of normal adult (92 micro g/g +/- 6 micro g/g), and proliferative scar (72 micro g/g +/- 5 micro g/g, all P < 0.01). The level of HA in wounded adult skin was significantly higher than that in the proliferative scar, and even much higher than that in normal skin (P < 0.01). The level of CD44 in normal fetal skin was significantly higher than that in proliferative scar and adult skin (all P < 0.01). The level of CD44 in wounded fetal skin 24 hours after injury decreased, significantly lower than that in normal fetal skin. There was no statistically significant difference between the level of CD44 in fetal skin one week after injury and that 24 hours after injury (P > 0.05). The level of CD44 in wounded adult skin was significantly higher than that in the normal adult skin (P < 0.01). The level of CD44 in the proliferative scar was between the level of CD44 in normal adult skin and that in wounded adult one. In normal fetal skin, CD44, positively stained at a moderate level, was distributed in keratinized cells, basic cells of hair follicle, and fibroblast of dermis. After injury, staining of CD44 became milder, especially by the incision. Immunohistochemistry showed that in normal adult skin, CD44 was distributed mainly in fibroblast of dermis and basic cells of hair follicle, weakly positively stained. After injury, the staning became stronger. CONCLUSION: The expression of HA and its receptor during the process of wound healing in human fetal skin is different from that in proliferative scars and adult skin, which might be one of the important causes of scarless healing of wounded fatal skin. PMID- 12899782 TI - [Advanced glycation end products stimulate human endothelial cells to produce monocyte chemoattractant protein-1]. AB - OBJECTIVE: To investigate the effects of advanced glycation end products (AGE) on secretion of monocyte chemoattractant protein-1 (MCP-1) by human endothelial cells and its signal transduction pathway. METHODS: Human umbilical vein endothelial cells (HUVECs) and HUVEC-derived cell line (ECV304) were cultured in vitro with indicated concentration of AGE modified human serum albumin (AGE-HSA) or AGE modified bovine serum albumin (AGE-BSA). The production of MCP-1 was evaluated by Western blotting and enzyme-linked immunoadsorbent assay (ELISA). The MCP-1 mRNA expression was assayed by reverse-transcription polymerase chain reaction (RT-PCR). Intracellular oxidative stress was detected by flow cytometry. The phosphorylation activity of cellular p38 mitogen-activated protein kinase (p38-MAPK) was analyzed by Western blotting using a phospho-specific antibody. RESULTS: AGE-HSA and AGE-BSA, but not their unmodified form, upregulated the expression of MCP-1 mRNA and protein dose- and time-dependently. The MCP-1 concentration in the supernatant of HUVECs incubated with 50 micro g/ml AGE-HSA for 12 hours increased from 48.3 pg/ micro g +/- 0.6 pg/ micro g protein to 148.1 pg/ micro g +/- 12.6 pg/ micro g protein (P < 0.01). AGE modified proteins were associated with enhanced oxidative stress and p38-MAPK phosphorylation activity. Incubation of HUVECs with 50 micro g/ml AGE-HSA for 30 minutes resulted in increase of p38-MAPK phosphorylation activity by 91% +/- 14% (P < 0.01). Antioxidant or SB 203580, a specific inhibitor of p38, could block the over expression of MCP-1. CONCLUSION: AGE modified proteins stimulate endothelial cells to produce MCP-1 through activation of the p38 signal pathway. This effect may contribute to the pathogenesis of atherosclerosis seen in AGE-associated diseases. PMID- 12899783 TI - [Prediction synthesis and identification of HLA-A2-restricted cytotoxic T lymphocyte epitopes of the tumor antigen MAGE-n]. AB - OBJECTIVE: To predict, synthesize, and identify HLA-A2-restricted cytotoxic T lymphocyte (CTL) epitopes of the tumor the novel antigen MAGE-n. METHODS: Long distance prediction system SYFPEITHI combined with polynomial method was used to predict the HLA-A2-restricted CTL epitopes of the tumor antigen MAGE-n. The candidate epitopes were synthesized with solid phase strategies, purified with reverse phase high-performance liquid chromatography and identified by mass spectrometry, The binding affinity and biding stability of the synthesized peptides were examined by cellular competition-based HLA-A2 peptide binding assay, T2 peptide stabilization assay, and peptide-major histocompatibility complex dissociation assay. RESULTS: Five HLA-A2-restricted CTL epitopes of MAGE n were selected: The epitopes QLVFGIEVV (159-167), IMPKTGGLI (195-203), and FLIIVLMI (201-209) with high HLA-A2 binding affinity (LC(50) < 15 micro mol/L) and binding stability (DT(50) > 6 h) were selected as candidate epitopes for further study in immunotherapy for tumor. CONCLUSION: Epitope prediction combined with epitope reconstruction improves the study of HLA-A2-restricted CTL epitopes of the tumor antigen MAGE-n. The selected epitopes of MAGE-n may be used in the design of therapeutic peptide vaccine for hepatocellular carcinoma. PMID- 12899784 TI - [Interaction effects between 1,25-dihydroxyvitamin D3 and transforming growth factor-beta1 on the proliferation and differentiation of human embryonic osteoblasts]. AB - OBJECTIVE: To characterize the interaction effects between 1,25-dihydroxyvitamin D(3) [1,25(OH)(2)D(3)] and human transforming growth factor-beta1 (hTGF-beta1) on the proliferation and differentiation of human embryonic osteoblasts. METHODS: Human embryonic osteoblasts were cultured. 1,25(OH)(2)D(3), of the concentration of 50 nmol/L, hTGF-beta1 of different concentrations (1 x 10(-8) g/L, 1 x 10(-7) g/L, 1 x 10(-6) g/L, and 1 x 10(-5) g/L), and both hTGF-beta1 of different concentrations and 1,25(OH)(2)D(3) of the concentration of 50 nmol/L were added into the cultures with a stimulation time of 96 hours. 3-(4, 5-dimethylthiazol-2 yl) - 2, 5 - diphenyl - tetrazolium bromide (MTT) was added into the cultures. ELISA technique was used to measure the optical density values so as to observe cell's number and proliferation. Alkaline phosphatase (ALP) activity was tested by King's method. Osteocalcin (OC) was tested by radioimmunoassay. The mRNA expression level of SMAD proteins, the key proteins of the TGF-beta signal transduction passageway, was detected by reverse transcriptase polymerase chain reaction (RT-PCR). RESULTS: MTT staining showed an absorbance of 0.086 +/- 0.022 in the 1,25(OH)(2)D(3) group, significantly lower than that of the control group (0.124 +/- 0.031, P < 0.05). The ALP activity in the culture of 1,25(OH)(2)D(3) group, hTGF-beta1 groups, and combined 1,25 (OH)(2)D(3) and hTGF-beta1 groups were 1.3 - 2.0 times that of the control group (all P < 0.05). When the concentration of hTGF-beta1 is 1 x 10(-6) g/L, an interaction effect was found between 1,25(OH)(2)D(3) and hTGF-beta1 on the ALP activity of human embryonic osteoblasts. The OC value in cell medium was 5.3 micro g/L +/- 1.6 micro g/L and 5.4 micro g/L +/- 0.9 micro g/L respectively in the hTGF-beta1 1 x 10(-6) g/L and 1 x 10(-5) g/L combined with 1,25(OH)(2)D(3) (50 nmol/L) groups, significantly higher than that in the control group (P < 0.05), but no interaction effect was found between 1,25(OH)(2)D(3) and hTGF-beta1 on the OC production. hTGF-beta1 of different concentrations and hTGF-beta1 of different concentrations combined with 1,25(OH)(2)D(3) all increased the SMAD3 mRNA expression level in human embryonic osteoblasts (all P < 0.05). When the human embryonic osteoblasts were treated with 1,25(OH)(2)D(3) combined with 1 x 10(-6) g/L hTGF-beta1, its SMAD3 mRNA level reached the peak, about 6-fold that of the control group. 1,25(OH)(2)VD(3) and/or hTGF-beta1 did not influenced the SMAD4 mRNA expression. CONCLUSION: 1,25(OH)(2)D(3) inhibits the proliferation of human embryonic osteoblasts and increases the expression of osteoblastic markers, such as ALP and OC, thus promoting its differentiation. An interaction effect exists between 1,25(OH)(2)D(3) and hTGF-beta1 on the differentiation of human embryonic osteoblasts, which may be partly induced by SMA D(3) protein in the TGF-beta signal transduction passageway. PMID- 12899785 TI - [The association of cyclin D1 (A870G) polymorphism with susceptibility to esophageal and cardiac cancer in north Chinese population]. AB - OBJECTIVE: To investigate the association of Cyclin D1 (A870G) single nucleotide polymorphism (SNP) with the susceptibility to esophageal and cardiac cancer in northern Chinese population. METHODS: By polymerase chain reaction-single strand conformation polymorphism analysis (PCR-SSCP), the Cyclin D1 (A870G) genotyping was performed among 178 patients with esophageal or esophageal -gastric junction carcinoma (120 with esophageal squamous cell cancer and 58 with cardiac adenoma cancer) and 122 health controls. RESULTS: There were no significant differences in Cyclin D1 (A870G) allele frequencies between cancer patients and health controls (P = 0.075). There is no genotype distribution difference was found between non-smoker patients and controls (P > 0.05). The risk for esophageal and cardiac cancer is 2.5 times higher in A/A genotype carried smokers than that in A/G or G/G genotype carried non-smokers, with the adjusted Odd Ratio of 2.57 (95% confidence interval is 1.19 approximately 5.57). The G/G genotype reduces the susceptibility to esophageal cancer, with the adjusted Odd Ratio of 0.39 and 95% confidence interval of 0.18 - 0.82. The A/A frequency in cardiac patients (34.48%) is higher than that in health controls (23.77%) but the difference does not reach significant (P = 0.212). CONCLUSION: In northern Chinese population, the smoking individuals carrying Cyclin D1 (A870G) A/A genotype increase the susceptibility to esophageal and cardiac cancer. The G/G genotype probably plays a protecting role in the occurrence of esophageal cancer. PMID- 12899786 TI - [Clinical manifestation, treatment, and outcome of severe acute respiratory syndrome: analysis of 108 cases in Beijing]. AB - OBJECTIVE: To investigate the clinical manifestations, treatment, and outcome of severe acute respiratory syndrome (SARS). METHODS: The clinical data of 108 SARS in-patients were analyzed. RESULTS: Among the 108 cases, 35 males (32.4%) and 73 females (67.6%), aged 37 +/- 9 (range 13 approximately 83 years), most were white collar workers, medical workers accounting for 28.7%. 87.0% had a definite contact history. 20.4% were complicated by other internal diseases. The main clinical manifestations included fever, cough, pectoralgia, chest distress, headache, etc. Involvement of multiple organs was often. The incubation period was 2 approximately 14 days. The course included early stage, progressive stage, climax, and convalescence. Mild type accounted for 4.6%, common type 39.8%, severe type 20.4%, and extremely severe type 35.2%. At the early stage, white blood cell count, lymphocyte count and ratio of lymphocyte, proalbumin, transferring, CD(3)(+), CD(4)(+), CD(8)(+) cell count were remarkably decreased, and C-reactive protein, alpha-acid glycoprotein, and alpha(2)-globulin were remarkably increased. X-ray chest film showed solitary or multiple local exudative changes, mostly in pulmonary lower field. During the progressive stage the hyaline or consolidation images were enlarged. High solution CT showed solitary or multiple cotton wadding like images and ground glass-like or consolidation images. Hypoxemia was common. At the progressive stage application of adrenocortical hormones and non-invasive mechanical ventilation helped stop the progress of disease. Ninety-four cases were discharged, 14 cases died. Traditional Chinese medicine (TCM) regards SARS as one of epidemic febrile diseases. Treatment by combination of TCM and Western medicine was effective. CONCLUSION: SARS mainly affects youth and people in their prime of life. It has its own characteristic clinical manifestations. The classification system of 5 types and 4 stages helps judge the condition. Treatment by combination of TCM and Western medicine is encouraged. PMID- 12899787 TI - [The epidemiological findings and the experience in the prevention and control of SARS in the Hong Kong Special Administrative Region]. AB - OBJECTIVE: To describe the epidemiological findings and the experience in the prevention and control of SARS in the Hong Kong Special Administrative Region. METHOD: The outbreak of SARS in Hong Kong was reviewed. Relevant data was collected and epidemiological studies conducted. Preventive measures taken by the Government from the public health perspective were introduced. RESULT: Based on the results of epidemiological investigation, the incubation period of SARS typically ranges from 2 - 7 days, but can be over 10 day. Transmission of SARS is mainly through respiratory droplets, and contacts with objects contaminated with a patient's respiratory droplets and secretions. Most of the SARS patients are health care workers and close contacts of SARS patients. Outbreak control measures include: firstly, controlling the source of infection through early detection, early diagnosis, early reporting, early isolation and early treatment: secondly, break the chain of transmission: thirdly, protecting the vulnerable; and fourthly, dissemination of information. At the same time, specific measures which include health checks at border points, home confinement of contacts and school suspension were taken. All of these measures help contain the SARS outbreak. CONCLUSION: Prevention of infectious diseases calls for macro and integrated strategies as well as effective surveillance. Decisive and prompt actions are necessary to control epidemics. PMID- 12899788 TI - [Epidemiological study of the transmission chain of a severe acute respiratory syndrome outbreak]. AB - OBJECTIVE: To investigate the process of SARS transmission and to evaluate infectivity of SARS patients in different periods of disease development. METHODS: A case of SARS outbreak beginning from a male, 74-year-old patient in Beijing. Two hundred and seventy five primary, secondary and tertiary contacts, including family members relatives, colleagues, and others exposed to him, were traced by field investigation and telephone interview using a standardized questionnaire. The transmission process and infectivity of disease were analyzed with transmission chain diagram and contact history diagram. RESULTS: Among the 275 contacts of the index case, 41 fell ill of SARS with an attack rate of 14.9% and 5 deaths with a fatality rate of 12.2%. The transmission chain of this outbreak could be clearly delineated, and all the 41 cases had close contacts with their infectors, SARS cases of the previous generation within the symptomatic period thereof. All of the 114 contacts that contacted their infectors, SARS cases of the previous generation during the incubation period thereof remained healthy. The incubation period of 27 cases that had contacted their infectors only once was 1 - 10 days with a median of 3 days. The highest risk of being infected was caring the patients unprotectedly, followed by living together with patient, visiting patient, and handling patient without protection. CONCLUSION: All the secondary cases have the history of close contact with their symptomatic infectors of previous generation. It cannot be proved at all that SARS is infective during the incubation period. The infectivity of SARS is related to the degree of closeness of contact with the patient. PMID- 12899789 TI - [Clinical analysis of SARS: 27 cases report]. AB - OBJECTIVE: To realize the clinical characteristic of severe acute respiratory syndrome (SARS). METHODS: Observe the clinical signs and symptoms of each patients; do laboratory examination periodically; and at last do Stat analysis of all the cases. RESULTS: The first onset symptom of the 27 cases is high fever accompany with toxic symptom such as muscular soreness, headache, etc. The patient's condition may be most serious in the 2nd week. The amount of end-brush blood WBC decrease, especially the lymphocyte decrease more obviously. The amount of CD(4)(+) and CD(8)(+) decrease obviously is another important characteristic. The chest X-ray show pulmonary lesion in the 2nd day after onset of the illness, but 40.7% cases show positive change after 6 days onset. During the 7 - 12 days the pulmonary lesion become most serious. Seven cases are treated by steroid in the first 3 days when the chest X-ray is normal, but they become positive during 6 - 8 days of the course. Whether the state of the illness is serious or not seems no obvious relationship to when they are treated by steroid. And large dosage of steroid may inhibit the level of CD(4)(+) and CD8 (P < 0.05). Two severe cases present double infection after large dosage of steroid. CONCLUSION: The course of a infectious atypical pneumonia is almost 3 weeks, the 1st week can be decided as early stage, the 2nd week is decided as fastigium, the 3rd week is decided as stage of recovery. During the fastigium, the disease transform very quickly, we need pay more attention to the patient. The CD(4)(+), CD(8)(+) and CD(3)(+) of SARS patient decrease obviously, it seems the immuno-function inhibited seriously. The serious case is easily accompany with severe secondary infection in the late stage, so the use of steroid must be carefully and the dosage need not to be much more. PMID- 12899790 TI - [Antineutrophil cytoplasmic antibody associated vasculitis induced by antithyroid agents]. AB - OBJECTIVE: To investigate the significance of antineutrophil cytoplasmic antibody (ANCA) in hyperthyroidism at different stages and treated with different drugs and to investigate the relationship between the ANCA specific target antigens and the clinical manifestations of vasculitis. METHODS: Two hundred and sixteen patients with hyperthyroidism were divided into four groups: untreated (n = 34), treated with propylthiouracil (PTU) (n = 62), treated with methimazole (MMI) (n = 77), and treated with both PTU and MMI (n = 43). Sera were collected from the 216 patients. Indirect immunofluorescence (IIF) test was used to detect the ANCA and antinuclear antibody (ANA). Antigen-specific ELISA was used to detect the existence of 7 specific target antigens of ANCA: myeloperoxidase (MPO), proteinase 3 (PR3), lactoferrin (LF), human leukocyte elastase (HLE), azurocidin (AZU), cathepsinG (CG), and bactericidal/permeability-increasing protein (BPI) in the sera positive in ANCA and ANA determined by IIF. RESULTS: The IIF positive rate was 5.9% (2/34) in the untreated group; 22.6% (14/62) in the patients treated with PTU, all IIF-ANCA positive; 6.5% (5/77) in the patients treated with MMI, all IIF-ANA positive; and 27.9% (12/43) in the patients treated with both PTU and MMI, 8 IIF-ANA positive and 3 IIF-ANCA positive. The IIF-ANCA positive rate in the patients receiving PTU was significantly higher than that in the untreated patients (P < 0.017). The IIF-ANA and IIF-ANCA positive rates of the patients treated with PTU were both significantly higher than those of the patients treated with MMI (P < 0.017). Six of the IIF-positive 31 patients administered with drugs (19.4%) had signs and symptoms associated with vasculitis. Of the six patients, 4 were MPO antibody positive (66.7%), 2 were PR3 antibody positive (33.3%), 4 were LF antibody positive, 3 were HLE antibody positive, 3 were AZU antibody positive, and 2 were CG antibody positive. None was anti-BPI antibody positive. Two of the six patients were both anti-MPO and anti PR3 negative. All MPO-ANCA positive patients had clinical signs and symptoms of vasculitis. CONCLUSION: PTU is associated with the production of ANCA in patients with hyperthyroidism. PTU induced ANCA is caused by polyclonal activation of B cells. Anti-MPO antibody may be related to the occurrence of clinical vasculitis. PMID- 12899791 TI - [Correlation between serum thyroglobulin and thyroid stimulating hormone in populations with non-toxic goiter]. AB - OBJECTIVE: To investigate the correlation between serum thyroglobulin (Tg) and thyroid stimulating hormone (TSH) in populations with non-toxic goiter. METHODS: Thyroid ultrasonography was conducted, and determination of serum triiodothyronine (FT3), thyroxin (FT4), Tg, thyroid peroxidase antibody (TPOAb), thyroglobulin antibody (TGAb) and TSH, and urine iodine were carried out among 609 persons with an average age of 35.4 in Panshan county, a mild iodine deficient area in Liaoning province, and 1136 persons aged 39.1 on average in Zhangwu county, an iodine sufficient area in Liaoning province, totally 2 320 persons. RESULTS: The serum TSH levels of the populations with diffuse and nodular goiter in Panshan county were (0.11 +/- 0.08) mU/L and (0.90 +/- 0.10) mU/L, significantly higher than those of the normal population in the same areas (1.33 +/- 0.04) mU/L, both P < 0.01. The serum TSH levels of the populations with diffuse and nodular goiter in Zhangwu county were (1.29 +/- 0.13) mU/L and (0.89 +/- 0.14) mU/L, significantly higher than those of the normal population in the same areas (1.74 +/- 0.04) mU/L, both P < 0.01. The serum TSH level of normal population in iodine deficient area was significantly lower than that of normal population in iodine sufficient area (P < 0.01). The serum TSH level of the normal population in Panshan County was significantly lower than that of the normal population in Zhangwu County (P < 0.01). The serum Tg level of the populations with diffuse and nodular goiter in Panshan county were (12.0 +/- 1.21) ng/L and (50.1 +/- 12.20) ng/L, significantly higher than that of the normal population in the same area (9.1 +/- 2.44) ng/L, P < 0.05 and P < 0.01. The serum Tg level of the populations with diffuse and nodular goiter in Zhangwu county were (12.4 +/- 2.11) ng/L and (61.2 +/- 11.86) ng/L, significantly higher than that of the normal population in the same area (7.7 +/- 2.3) ng/L, P < 0.05 and P < 0.01. The Tg level of the normal population in Panshan County was significantly higher than that of the normal population in Zhangwu county (P < 0.01). The Tg level was positively correlated with the thyroid volume in the nodular goiter population. CONCLUSION: The occurrence of goiter is not directly related to TSH stimulation in mild iodine deficient area. Nontoxic goiter shows a thyroid autonomic function. The major cause of high serum Tg in nodular goiter may be the leakage of Tg into blood circulation caused by degeneration of large colloid follicles and destruction of follicular architecture, not the volume increase of thyroid and its autonomic function. PMID- 12899792 TI - [Significance of combining tests of thyroid autoantibodies in differential diagnosis between Graves's disease and Hashimoto's hyperthyroidism]. AB - OBJECTIVE: To investigate the differential diagnosis between Graves's disease and Hashimoto's hyperthyroidism to improve positive diagnostic rate in Hashimoto's hyperthyroidism, to facilitate proper management. METHODS: Cytological examination was conducted by fine needle biopsy (FNB) on the thyroids of patients with hyperthyroidism diagnosed by clinical data, including 22 cases of Hashimoto's hyperthyroidism, 5 males and 17 females, aged 39 +/- 12, and 49 cases of Graves's disease, 20 males and 29 females, aged 41 +/- 11. Thyroid auto antibodies-thyroid peroxidase antibody (TPOAb), thyroid microsomal antibody (TMA), thyroglobulin antibody (TGA), thyroid-stimulating hormone receptor antibody (TRAb), and free serum triiodothyronine (FT3), free thyroxin (FT4), and serum thyroid-stimulating hormone (S-TSH) were tested. The correlation between the pathology of thyroid and different autoantibodies and thyroid hormone was analyzed. RESULTS: TRAb was 42.28 +/- 75.89 in Graves disease group, significantly higher than that in Hashimoto's hyperthyroidism group (5.77 +/- 12.69, P < 0.001). FT3 was 16.13 +/- 9.95, significantly higher than that in Hashimoto's hyperthyroidism group (9.54 +/- 6.76, P < 0.01). In Graves's disease group the positive rates for TRAb, TPOAb, TMA, and TGA were 83.67%, 75.51%, 57.14% and 48.98% respectively. In Hashimoto's hyperthyroidism group, the positive rate for TRAb was 13.64%, and the positive rates for the rest autoantibodies were all 86.36%. The double positive rate of TPOAb and TGA was 86.36% in Hashimoto's hyperthyroidism group, significantly higher than that in Graves's disease group (48.98%). There was a positive association between TRAb and FT3, and between diffuse and symmetric increased size of thyroid and FT3 TRAb. CONCLUSION: Comprehensive analysis of TRAb, TPOAb, TGA, and FT3 is helpful to the differential diagnosis between Graves's disease and Hashimoto's hyperthyroidism. PMID- 12899793 TI - [Clinical study of anti-hepatic fibrosis effect of IFN-gamma in patients with chronic hepatitis B]. AB - OBJECTIVE: This was an open, random, control and multicenter clinical trial. In the study, the anti-hepatic fibrosis effect of rhIFN-gamma and its side reactions were evaluated. METHODS: A total of 289 patients enrolled in clinical trial, 153 patients in trial group and 136 patients in control group. The routine strategy is given in all patients. In addition, every patient in trial group received rhIFN-gamma at a dose of 1 MU intramuscularly daily for the first three months and 1 MU every other day for the following six months. Patients of two groups were followed up for another three months after the treatment. Histological indices, serum hepatic fibrosis indices, ultrasound B and symptoms and signs evaluated the effect. For patients who accepted two liver biopsies before and after treatment, the effect was determined by semiquantitative score system while for patients who did not accepted liver biopsy, serum hepatic fibrosis indices and ultrasound B were major criteria. RESULTS: The efficient of treatment were 66% in trial group vs. 16.2% in control group and the obviously efficient of treatment was 27.8% in trial group vs. 7.4% in control group (P < 0.001). For patients who accepted two liver biopsies before and after treatment, the efficient of treatment was 63% in trial group vs. 24.1% in control group and the obviously efficient of treatment was 27.8% in trial group vs. 13.8%. For patients who did not accepted liver biopsy, the efficient of treatment was 67.7% in trial group vs. 14.0% in control group and the obviously efficient of treatment was 22.2% in trial group vs. 5.6%. None of patients emerged serious side reactions during clinical trial. CONCLUSION: The results confirmed that rhIFN-gamma have a better anti-hepatic fibrosis effect to patients with chronic hepatitis B. PMID- 12899794 TI - [Treatment of malignant lymphoma by autologous hematopoietic stem cell transplantation combined with high dose radiotherapy and chemotherapy]. AB - OBJECTIVE: To investigate the effects of treatment of malignant lymphoma (ML) by autologous hematopoietic stem cell transplantation (AHSCT) combined with high dose radiotherapy and chemotherapy. METHODS: From June 1991 to October 2002, 56 patients, aged 9 - 65 years, with ML, 44 non-Hodgkin's lymphoma (NHL) and 12 Hodgkin's disease (HD) were treated with AGSCT combined with high dose radiotherapy and chemotherapy. Among them, 12 received autologous bone marrow transplantation (ABMT) and 44 received autologous peripheral blood stem cell transplantation (APBSCT), In the latter the APBSCs were mobilized by carboplatin and etoposide (CE) or CHOP (CTX, ADM, VCR, PDN) plus granulocyte-colony stimulating factor (G-CSF) and/or granulocyte-macrophage colony stimulating factor (GM-CSF) 10 micro g/kg/d. Pretreatment included BEAM (BCNU, Vp-16, AraC, MEL) or MEL 140 mg/m(2) + Vp-16,200 mg/L total body irradiation (TBI) 8 Gy. The patients were flowed up for 40 - 4,310 days with a median of 1,820 days. RESULTS: Hematopoietic reconstruction was observed in all patients. The 1, 2, and 5-year disease-free survival rates were 87.5%, 60.7%, and 57.1% respectively, with the longest survival of 11 years. Fourteen patients developed relapse. No transplantation-related death was seen. CONCLUSION: Autologous hematopoietic stem cell transplantation combined with high dose radiotherapy and chemotherapy is effective on Ml. AHSCT shows a more rapid effect on hematopoietic reconstruction than ABMT. PMID- 12899795 TI - [Application of diffusion-weighted and perfusion magnetic resonance imaging in definition of the ischemic penumbra in hyperacute cerebral infarction]. AB - OBJECTIVE: To investigate the value of diffusion-weighted imaging (DWI) and perfusion imaging (PI) in definition of the ischemic penumbra in hyperacute cerebral infarction. METHODS: (1) Animal study: Fifty-five SD rats were randomly divided into 5 groups of 10 rats. Nylon monofilament was inserted into the left middle cerebral artery (MCA) and immediately pulled out in 10 rats as controls (sham operation group or group A). The MCA of 40 rats was occluded by nylon monofilament for 30 minutes (group B), 60 minutes (group C), 180 minutes (group D), and 360 minutes (group E). Then all rats were examined by T(1)WI, T(2)WI, DWI, and PI. Apparent diffusion coefficient (ADC), regional cerebral blood volume (rCBV), relative cerebral blood flow (rCBF) and mean transit time (MTT) topographical maps were archived by the Workstation (GE SUN ADW3.0). The relative values of ADC, rCBV, rCBF, and MTT (ratios between the values of the occluded side and the opposite side) were calculated. The rats were killed and the cerebral infarction areas were stained by TTC (C(19)H(15)N(4)CI) and observed by light microscopy and electron microscopy. The results of microscopic examination were compared with the DWI results. (2) Clinical study: Forty-three patients, 24 males and 19 females,aged 68 years on average with acute stroke symptoms underwent MRI examination within 6 hours of symptom onset. T(1)WI, T(2)WI, DWI and PI were performed on all patients at the first MRI examination and T(2)WI was performed for follow up. The relevant parameters, including ADC, rCBV, rCBF and MTT were computed. The values of ADC, rCBV, rCBF and MTT within the lesions were compared to those at the normal sides and the ratio was calculated. The lesion volumes were measured in ADC map (as vDWI), MTT map (as vPI) and follow-up T(2)WI (as vCI). The differences between vDWI and vPI, and between vDWI and vCI were compared. RESULTS: (1) Animal study: In the control group DWI and PI showed no change in the signal intensity and no pathological change was found. Hyper intensity signal was found in the T(2)WI in groups D and E. No abnormal signal was found in the T(1)WI in all groups. Abnormal signals were found in the MCA supplying region in the DWI and PI of groups B, C, D, and E. The size of hyper intensity signal in DWI along with the extension of occlusion time. TTC staining showed that the volume of unstained area was similar to that of hyper-intensity signal area in DWI image (t test, P > 0.05). PI showed a decrease of cerebral perfusion in the MCA-supplying region along with the extension of occlusion time. However, the size enlargement of occluded region was not as severe as shown by DWI along with the extension of occlusion time. The size of region with abnormal perfusion shown by PI was larger than that shown by DWI in groups B, C, and D. PI/DWI mismatch region, indicating the existence of ischemic penumbra zone, became gradually smaller along with the extension of occlusion time and almost disappeared 360 minutes after MCA occlusion. No significant change was found in the PI/DWI mismatch region by light microscope but slight swelling of mitochondrion and Golgi apparatus in neurons were observed by electron microscope. (2) Clinical study: The cases of 38 of the 43 patients were diagnosed as cerebral infarction by clinical and MRI follow-up examinations. The first MRI examination of these 38 cases showed no abnormal signal in T(1)WI and hyper intensity signal in T(2)WI of 4 cases. The vPI was larger than vDWI in 31 cases and the vPI was equal to or smaller than the vDWI in 7 cases. Follow-up examination showed that vCI was larger than vDWI in 28 cases and vCI almost equaled to vDWI in 10 cases. Spearman correlation analysis showed a positive correlation between the volume of ischemic penumbra zone and that of infarction (r = 0.689, P < 0.001) which suggests that the volume change of ischemic penumbra zone influences the size of final infarction directly. CONCLUSION: Combination of DWI and PI clearly defines the ischemic penumbra in hyperacute stroke patient, thus laying, thus laying a basis for therapeutic intervention plan for stroke patients. PMID- 12899796 TI - [Copper ion electrochemistry in treatment of bleeding and prolapse due to hemorrhoid, experimental and clinical studies]. AB - OBJECTIVE: To observe the effect of copper ion electrochemistry (CIE) in treatment of bleeding and prolapse due to hemorrhoid. METHODS: Bleeding was caused at several points on the sacrospinal muscles of 26 rabbits. Routine hemostasis was used at one side (control side) and CIE was used on the opposite side (experimental side). The stanch times at the different sides were recorded. Then the rabbits were killed. Pathological examination was made to the lung, liver, and kidney to observe the amount of small thrombus and inflammation. CIE was performed at the right side of anus of 6 dogs and the right side was used as control side. Tissue specimens of rabbit muscle and dog anus mucosa were collected to examine the copper ion concentration. A copper needle was inserted into each pile to the depth of 10 - 15 mm for 4'40" (with the mean number of treated points of 9.8) upon 202 patients suffering from bleeding due to hemorrhoid, 79 with inner hemorrhoids and 123 patients with mixed hemorrhoids (experimental group). Another 171 patients suffering from bleeding due to hemorrhoid, 64 with inner hemorrhoids and 107 with mixed hemorrhoids were treated with suppository as controls. CEI was performed on 128 patients suffering from prolapse of hemorrhoid, 41 with inner hemorrhoids and 87 with mixed hemorrhoids (experimental group). Another 115 patients suffering from prolapse of hemorrhoid, 40 with inner hemorrhoids and 75 with mixed hemorrhoids were treated with suppository as controls. Biopsy specimens of anus mucosa were taken from 18 patients with mixed hemorrhoid who underwent CIE. The copper ion concentration was measured in 30 patients undergoing CIE. RESULTS: The stanch time in the experimental side of rabbit was (1.16 +/- 0.18) min, significantly shorter than that in the control side [(2.13 +/- 0.46) min, P = 0.0037]. Extensive small thrombi and edema of vascular wall were seen in the rabbit liver, lung, kidney, and muscle tissues in comparison with the tissues of the control side (P < 0.01). Copper ion complex was seen in the experimental rabbit tissues. The copper ion concentration was significantly higher in the experimental area of dog anus mucosa than in the control area. Clinical study showed that after CIE the curative rate for bleeding was 95.5%, significantly higher than that of the control group (8.8%, U = 44.6, P < 0.001). The curative rate for prolapse in the experimental group was 60.8%, significantly higher than that of the control group (20.9%, U = 313.2, P < 0.01). No patient felt pain while treated and later. After the CIE treatment, the patients only needed to take a rest for 4 hours. Pathology showed there were much more small thrombi in the treated tissues. The blood copper ion concentration in the trial group did not increase significantly after CIE in comparison with that before treatment. CONCLUSION: CIE is safe, effective and easy to perform in treatment of bleeding and prolapse due to hemorrhoid. PMID- 12899797 TI - [The study of peri-operative chemotherapy in stage I-IIIa NSCLC]. AB - OBJECTIVE: A number of studies had evaluated the benefit of neoadjuvant chemotherapy combined surgery on stage IIIa-IIIb NSCLC, survival benefit was found in several papers. We attempt to evaluate the survival and prognosis of cisplatinum-based schedule as peri-operative CT for resectable stage I-IIIa NSCLC. METHODS: A prospective, randomized, multicenter study was conducted by Shanghai Lung Cancer Team (supported by Shanghai Branch of Discipline Foundation) since 1995-1997 for 211 cases of stage I-IIIa NSCLC with curative resection (99 stage I, 47 stage II, 65 stage III), age of or= 80, staged by 1997 AJC TNM Criteria. They were randomized to be 103 cases with 1 - 2 cycles of pre operative CT and 108 cases with no pre-operative CT, 2 - 4 cycles of post operative CT were used for stage II and stage IIIa NSCLC, it was totally 4 cycles of MVP or MOP CT schedule each case. Follow-up team had been trained, the follow up rate should be >or= 95%, last follow-up date was March of 2002. Lobectomy was performed for most patients. Accumulated survival, log rank, MST, Cox uni variance and multi-variance analyses were used as statistics for evaluation. RESULTS: The two arms were well balanced for baseline demographic and clinical characteristics (P > 0.05 for all). Stage I NSCLC had the best year-survival in whole patients. No statistical survival difference was found between the group with pre-op CT and with no pre-op CT, P = 0.074, 0.087 and 0.097, respectively, 5 year survival rates were of 31.98%:36.68%. In various stage, a statistical survival difference was only shown in stage IINSCLC, P = 0.042, 5-year survival rates and MST were worse in the group with pre-operation CT, 20%:65.2% and 24 months:48 months, respectively, but no difference was seen in stage I and stage IIIa NSCLC. Stage and post-operation CT were the only two meaningful parameters with statistical survival difference calculated by multi-variance analyses, P = 0.000 all, but no difference was found in others 4 parameters (age, sex, type and pre-operation CT). The response rate of pre-operation CT was of 50%. Though there was no statistical difference, the responders were with slightly better year survival rates than MR + NR patients, 38.9% and 33.3%, respectively. In the cases with pathological "T" down stage and "T" unchanged after pre-operation CT had a better yr-survival rates than "T" up-stage, P = 0.03, 5-year survival rates were of 41.67%, 40.51% and 11.76%, respectively, thus, effective chemotherapy might be beneficial to survival. Besides, in the cases with >or= 3 cycles of post operation CT have better survival rates than less cycles. CONCLUSION: A prospective, randomized, multicenter peri-operation CT study for stage I-IIIa NSCLC conducted in Shanghai, China., it showed there had no benefit in survival between with pre-operation CT arm and with no pre-operation CT arm. In stage II NSCLC, pre-operation CT cases had a worse year-survival than with no pre operation CT, P = 0.042, but no difference was seen in stage I and stage IIIa NSCLC. The responder of CT and "T" down stage, "T" unchanged had better survival rates than those of not response and "T" up-stage. From multivariate analyses, stage and post-operation CT were the two meaningful parameters to year-survival, >or= 3 - 4 cycles of post-operation CT had a better statistical higher year survival than less cycles. Nutrition, supportive treatment, immunity status and prevention of toxicity might be the next study worthy to conduct, for CT combined with OP. PMID- 12899798 TI - [Integrin alpha3beta1 mediates hepatocellular carcinoma cell adhesion and chemotaxis to type IV collagen]. AB - OBJECTIVE: To investigate the effects of Integrin alpha(3)beta(1) on the adhesion and chemotaxis of hepatocellular carcinoma (HCC) cells to type IV collagen (Col IV). METHODS: (1) HCC cells were culture and suspension of HCC cells was made. Anti-alpha(3) and Anti-beta(1) were added into the HCC cell suspension. Flow cytometry was used to determine the expression of integrin alpha(3)beta(1) on the surface of HCC. (2) 5 micro g/ml Col IV was used to coat a cell with the diameter of 25 mm. Digested HCC cells were added. Anti-alpha(3) and Anti-beta(1) of the concentrations of 5 micro g/ml and 10 micro g/ml respectively were added into the cell suspension. Before and after the addition of Anti-alpha(3) and Anti-beta(1), micropipette technique was used to measure the adhesion force of HCC on Col IV coated surface, as function of the square of internal radius of micropipette and the critical negative pressure needed to detach a single HCC cell away from the substrate. (3) Col IV of the concentration of 600 micro g/ml was added into the dual micropipettes. Then the dual micropipettes were led towards the HCC cells. A HCC cell was made to seal the openings of the 2 micropipettes with different parts of the cell contacting Col IV in different micropipettes. The pseudopod protrusion was observed dynamically and recorded with tape recorder. The length of pseudopod was measured and plotted against the chemotactic time so as to obtain a pseudopod growth curve. RESULTS: (1) The expression rates of integrin subunit alpha(3) and beta(1) on the surface of HCC cells were 95.55% and 95.78% respectively. (2) The adhesion force of HCC cells to the 5 micro g/ml Col IV coated surface was 932 +/- 134 (x 10(-10) N, n = 60). Upon treatment of the HCC cells with Anti-alpha(3) of the concentrations of 5 micro g/ml and 10 micro g/ml, the adhesion force decreased by 42% and 49%, to 536 +/- 122 (x 10(-10) N, n = 60) and 476 +/- 63 (x 10(-10) N, n = 60) respectively. Upon treatment of the HCC cells with Anti-beta(1) of the concentrations of 5 micro g/ml and 10 micro g/ml, the adhesion force decreased by 52% and 76%, to 449 +/- 119 (x 10(-10) N, n = 60) and 220 +/- 78 (x 10(-10) N, n = 60) respectively. (3) The length of pseudopod increased along with the chemotactic time. The pseudopod length and growth curve were almost identical in the dual micropipettes when they were filled with Col IV. When Anti-alpha(3) or Anti-beta(1) was added into one of the dual micropipettes, the HCC cell pseudopod protrusion was almost blocked completely, while the HCC cell pseudopod in the opposite micropipette became more evident. CONCLUSION: Integrin alpha(3)beta(1) is an important constituent receptor in mediating HCC cell adhesion and chemotactic pseudopod protrusion to Col IV. PMID- 12899799 TI - [Ovarian carcinoma cell inhibits T cell JAK-STAT signal transduction pathway, an experimental study]. AB - OBJECTIVE: To investigate the effect of ovarian carcinoma cell on T cell JAK-STAT signal transduction pathway and its role in the ovarian carcinoma induced immunosupression. METHODS: Human ovarian carcinoma cells of OVCAR3. CAOV3, and SKOV3 lines were cultured. CD8(+) T cells were isolated from the peripheral venous blood of healthy persons. Then the supernatants of these ovarian carcinoma cell lines and RPMI-1640 were added into the culture of CD8(+) T cells (groups I, II, III, and control). Thiazolyl blue (MTT) method was used to detect the growth of CD8(+) T cell. The cell cycle was examined by flow cytometry. The secretion of the Tc1 type cytokine interferon (IFN)-gamma mRNA and the secretion of the Tc2 type cytokine interleukin (IL)-10 mRNA were detected by RT-PCR. The expression of signaling molecules JAK and JAK3 and the phosphorylated activation of STAT3 and STAT5 in the CD8(+) T cell were analyzed by Western blotting. RESULTS: The absorbance at the wavelength 570 nm of CD8(+) T cell culture was 0.23 +/- 0.03, 0.28 +/- 0.06, and 0.29 +/- 0.05 in the group I, II, and III, all significantly lower than that in the group IV (0.79 +/- 0.07, all P < 0.01). The percentages of CD8(+) T cells at the stage S and stage G(2)/M were lower, and those in stage G(1)/G(0) were higher in groups I, II, and III than in group IV (all P < 0.01). The IFN-gamma expression was significantly lower in groups I, II, and III in comparison with that in group IV. However, the expression of IL-10 was significantly higher in groups I, II, and III in comparison with that in group IV. The expression of JAK3 protein, but not JAK1 protein, was significantly lower in groups I, II, and III in comparison with that in group IV. The phosphorylated activation of STAT5 was suppressed significantly in groups I, II, and III, whereas the phosphorylated activation of STAT3 was suppressed only in group I. CONCLUSION: Ovarian carcinoma may suppress T cell proliferation through inhibition of the JAK-STAT signal transduction pathway, which may be a mechanism of ovarian carcinoma induced immunosuppression. PMID- 12899800 TI - [Expression of aFGF in ovarian epithelial cancer and its signal transduction pathway]. AB - OBJECTIVE: To explore the expression of acidic fibroblast growth factor (aFGF) and its receptor FGFR1 in ovarian epithelial cancer and observe the effects of aFGF and TPK inhibitor Genistein on intracellular PKC and ERK activity in ovarian epithelial cancer cells line CAOV3. METHODS: The expression levels of aFGF and FGFR1 were evaluated by RT-PCR and western blot in 40 cases of ovarian epithelial cancer. The activity of PKC and ERK in cells induced by different concentration of aFGF and Genistein were detected by incorporation of [gamma-(32)P]-ATP into exogenous substrate. RESULTS: The expression levels of aFGFmRNA and FGFR1mRNA in the ovarian epithelial cancer were 0.981 +/- 0.130 and 1.047 +/- 0.148, respectively. Compared with normal ovary, ovarian tumor like condition and benign ovary tumors, the difference was significant (P < 0.05). The expression levels in stage III - IV were significantly higher than those in stage I - II (P < 0.05). There were overexpression of aFGF and FGFR1 in the ovarian epithelial cancer in western blot, too. The intracellular PKC and ERK activity increased with aFGF in a dose dependent manner, Genistein suppressed the intracellular PKC and ERK activity also in a dose dependent manner. CONCLUSION: aFGF may play an important role in carcinogenesis, development and invasion of ovarian epithelial cancer. Its receptor in human ovarian cancer cell line CAOV3 possessed TPK activity. These tyrosine-specific protein phosphorylation may initiate a cascade of biochemical events, which may increase the intracellular PKC and ERK activity. PKC and ERK locate downstream of TPK in CAOV3 cell line. PMID- 12899801 TI - [Infection of Coxsackievirus group B type 3 regulates the expression profile of chemokines in myocardial tissue/cells]. AB - OBJECTIVE: To investigate the role of Coxsackievirus group B type 3 (CVB3) infection on the expression profile of chemokines (ChKs) in myocardial tissue/cells. METHODS: CVB3 was inoculated into male BALB/c intraperitoneally and primary neonatal myocardial cells of BALB/c to establish CVB3 infection models in vivo and in vitro, where the expression profile of ChKs was detected at different time points post-infection as well as under different loading of CVB3 qualitatively and quantitatively by RT-PCR. RESULTS: The expression of MIP-2 and IP-10 was induced post-infection, while SDF-1, MCP-1, MCP-2, MCP-3, MCP-5, MDC, FKN and Ltn were constitutively expressed in myocardial tissue. The expression of MCP-1, MCP-2, MCP-3, MCP-5, MDC and Ltn increased 1.8, 1.9, 3.7, 1.7, 1.3 and 1.2 folds post-infection higher than that of uninfected control (P < 0.01). There was not significant difference in the expression of SDF-1 and FKN between infected myocardial tissue and uninfected myocardial tissue (P > 0.05). The expression of Eot was not detected in infected and uninfected myocardial tissue. Every chemokine had different expression at different infection time points. For example, the expression of MIP-2 at the 4th day was 1.1 and 1.5 times than that of the 7th and 14th day (P < 0.01). IP-10 showed similar expression between the 4th and 7th days (P > 0.05), which is 2.47 and 2.54 times compared to that of the 9th day (P < 0.01). And the expression of MCP-1 at the 14th day post-infection was 1.3, 1.2 and 1.0 times comparing to that of the 4th, 7th, 9th day post infection, which showed statistical meaning. The expression of MCP-2 at the 4th was 1.4, 1.5 and 2.2 times comparing to that of the 7th, 9th, 14th day, and moreover, the expression was lower than that of the basal expression. The expression of MCP-1 and MCP-3 was up-regulated significantly, which occurred at different time points post-infection in vitro. While the expression of MIP-2 and MCP-5 was down-regulated in vitro. The expression patterns of MCP-2, MCP-3, MCP-5 and MDC were consistent with CVB3 loading. But that of the others (FKN, SDF-1, et al) were inconsistent with CVB3 loading. There was a correlation between change patterns of MCP-3 and CVB3 loading post-infection (r = 0.881, P < 0.05) within 14 days after infection. The varied expression trend of MCP-1 and MCP-3 was similar to the titer of anti-CVB3 antibody (r = 0.913, P = 0.031), while the expression of MCP-2, MCP-5 and MDC shows contrary change. There was not a significant correlation between change patterns of other ChKs (IP-10, SDF-1, et al) and the titer of anti-CVB3 antibody. A positive correlation between anti-CVB3 antibody and MCP-1 (r = 0.976, P < 0.05) was showed. CONCLUSION: The expression level and kind of ChKs in vivo and in vitro was varied significantly in clusters after CVB3 infection. The ChKs changed in clusters consisted of the expression profiles of ChKs. There were complexity and unbalance in the change of the expression of ChKs in every expression profile. It suggested that CVB3 infection could regulate the expression of ChKs in myocardial tissue/cells likely in different ways. PMID- 12899802 TI - [Effect of zinc phthalocyanine-mediated photodynamic therapy on bone marrow purging, an experimental study]. AB - OBJECTIVE: To probe into the purging effects of zinc phthalocyanine-mediated photodynamic therapy (PDT) on simulated remission bone marrow grafts of chronic granulocytic leukemia. METHODS: (1) K562 cells, aline chronic granulocytic leukemia cells, and normal mononuclear cells (MNC) were cultured. Zinc phthalocyanine (ZnPcS(2)P(2)), a photosensitizer, with the terminal concentration of 1.0 micro g/ml was added into the cultures. The K562 cells and normal MNCs in the suspensions were broken. Fluorescence spectrophotometry was used to determine the concentration of zinc phthalocyanine in cells at different time points so as to find the optimal time for photodynamic purging process. (2) Suspensions of K562 cells and MNCs were made and incubated with zinc phthalocyanine of different concentrations (0.062 5, 0.125, 0.25, 0.5, and 1.0 micro g/ml) for 5 hours. A blank control group (sodium chloride of the same volume was added), a PDT control group (without photosensitizer), and a photosensitizer control group (zinc phthalocyanine was added without PDT) were established. Then the suspensions were irradiated with 670 nm laser. Trypan blue dye exclusion technique was used to calculate the number of live cells for a period of 5 days. The proliferative potency of K562 cells was detected by MTT colorimetric assay. The OD value was detected with ELISA apparatus to calculate the inhibition rate. Colony formation of K562 cells and MNCs was determined. (3) K562 cells were mixed into normal MNCs at the ratios of 1:100 and 1:1,000 so as to create the model of simulated remission bone marrow. After PDT treatment, colony formation test was done and nested-PCR was used to detect the bcr-abl mRNA expression in K562 cells. Colony formation test was made on the MNCs treated with PDT. The antiproliferative effects of PDT on normal hematopoietic progenitors were evaluated by CFU-Mix, CFU GM and CFU-E assays. RESULTS: (1) The zinc phthalocyanine content in the MNCs reached its peak within the first hour of incubation and then rapidly decreased to the lowest value in 4 hours. However, the zinc phthalocyanine content in the K562 cells gradually increased within the first 4 hours of incubation and reached its peak by the fifth hour with a ratio of zinc phthalocyanine content in K562 cells to that in MNCs of 4.59. Therefore, the fifth hour after incubation was selected as the optimal time to irradiate the suspensions using the laser with a wavelength of 670 nm. (2) The inhibitory rate of laser on the colony information rate was 91.1% for the K562 cells, 18.0% for the MNCs in CFU-Mix methyl cellulose culture system, 18.6% for the MNCs in CFU-GM methyl cellulose culture system, and 17.8% for the MNCs in CFU-E methyl cellulose culture system At the concentration of 0.25 micro g/ml, K562 cells were inhibited by 91.1%, however, CFU-Mix, CFU-GM and CFU-E were relatively spared, inhibitory rate being 18.0%, 18.6% and 17.8% respectively. (3) At the concentration of 0.25 micro g/ml, residual K562 cells in the simulated remission bone marrow were completely photoinactivated. CONCLUSION: Zinc phthalocyanine -based PDT selectively kills K562 cells. It would be a promising purging technique for chronic granulocytic leukemia. PMID- 12899803 TI - [Orexin A and neuropeptide Y levels in plasma and hypothalamus of rats with chronic renal failure]. AB - OBJECTIVE: To explore the changes of orexin A and neuropeptide (NPY) in plasma and hypothalamus of rats with chronic renal failure (CRF). METHODS: 41 male Wister rats weighing 200 approximately 250 g were randomly divided into three groups: normal group, sham operation group, and CRF group (with the right kidney and 2/3 of the left kidney resected). A certain number of rats were decapitated 4, 8,and 12 weeks after respectively. Their hypothalami were removed and blood collected. Radioimmunoassay was used to measure the levels of orexin A and NPY in hypothalamus and plasma. Automatic biochemical analyzer was used to measure the serum creatinine. RESULTS: The serum creatinine level of CRF rats was both significantly higher than those of the sham operation rats at week 8 and week 12, respectively. The plasma orexin A level of CRF rats at week 12 was 264 pg/ml +/- 62 pg/ml, significantly higher than that of sham operation group (183 pg/ml +/- 56pg/ml, P = 0.039). The hypothalamus orexin A level of CRF rats were 10.5 fmol/mg +/- 2.7 fmol/mg wet weight at week 12, significantly lower than that of sham operation rats (17.4 fmol/mg +/- 3.9 fmol/mg wet weight, P = 0.023). The plasma NPY levels of CRF rats at week 8 and week 12 were significantly higher than those of the sham operation rats (7.1 pmol/ml +/- 1.7 pmol/ml vs 5.0 pmol/ml +/- 0.5 pmol/ml, P = 0.01; and 7.9 pmol/ml +/- 1.1 pmol/ml vs 4.8 pmol/ml +/- 1.1 pmol/ml, P = 0.0008). The hypothalamus NPY level of CRF rats at week 12 were 70 fmol/mg +/- 23 fmol/mg wet weight, significantly lower than that of the sham operation rats (113 fmol/mg +/- 31 fmol/mg wet weight, P = 0.03). CONCLUSION: Loss of renal function may diminish the excretion of orexin A and neuropeptide. The lowering of hypothalamus orexin A and neuropeptide Y levels may be one of the causes inducing anorexia in CRF. PMID- 12899804 TI - [Effects of vascular endothelial growth factor and basic fibroblast growth factor in early stage diabetic retinopathy and their molecular pathological mechanism]. AB - OBJECTIVE: To study the effects of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) in early stage diabetic retinopathy and their mechanisms, so as to guide the clinical work theoretically. METHODS: Fifty five Wistar rats were randomly divided into four groups. Ten rats were used as controls (M group). Streptozocin was injected intraperitoneally to induce diabetes for I month (M(1) group, n = 15), 3 months (M(3) group, n = 15), and 5 months (M(5) group, n = 15). At the experimental ends of each group, the rats were over-anesthetized and their eyeballs were extracted to make digest preparation. In situ hybridization and immunohistochemistry were used to investigate the expression of bFGF and VEGF on retinal vascular. Transmission electron microscopy was used to observe the histology of the retinal vessels. RESULTS: There was no difference in the number of pericytes between M(1) group and M group (P > 0.05), but the number of pericytes was significantly lower in M(3) and M(5) groups than in the M and M(1) groups (P < 0.001). VEGF in situ hybridization showed an expression rate of 34% in the M(5) group. VEGF immunohistochemistry showed an expression rate of 56% in M(5) group. bFGF in situ hybridization showed an expression rate of 78% in M(3) group and an expression rate of 89% in M(5) group. Transmission electron microscopy showed no change in M group. However, it showed swelling of endothelial cells, finger-like process into the capillary cavity, and uneven distribution of heterochromatin in pericytes in M(1) group. Obvious fragmental thickening and splitting of basement membrane, swelling and deformation, finger like process to the capillary cavity, and concentration and margination of heterochromatin in endothelial cells and swelling and deformation of rough surfaced endoplasmic reticulum and mitochondrion in pericytes were seen in the M(3) and M(5) groups, especially the latter. CONCLUSION: During the course of diabetic retinopathy, morphologic changes of vessels occurs prior to the expression of growth factors, in which the expression of VEGF follows the expression of bFGF. PMID- 12899805 TI - Suicide risk not increased with SSRI antidepressants. AB - Depressed adult patients are no more likely to commit suicide while taking selective serotonin reuptake inhibitors (SSRIs) than any other class of antidepressants. The low suicide rate in the non-SSRI category, which included tricyclic antidepressants (TCAs), reminds us that no evidence exists to consider tricyclics more dangerous than other antidepressants, despite what is commonly believed. Thus, clinicians who avoid prescribing any antidepressant because of risk of suicide may change their clinical practice. It is important to remember that these patients are adults and had mild to moderate depression without any significant comorbid conditions. PMID- 12899806 TI - Low-dose warfarin prevents recurrent thromboembolism. AB - Low-intensity warfarin (target international normalized ratio [INR], 1.5-2.0) effectively prevents recurrent venous thromboembolism without increasing the risk of major bleeding when used long-term for secondary prophylaxis. This is a reasonable approach following at least 3 to 12 months of full-intensity warfarin after the initial thromboembolic event. PMID- 12899807 TI - Steroids ineffective for pain in children with pharyngitis. AB - In children with acute pharyngitis, oral dexamethasone does not provide clinically significant reductions in time to initial or complete pain relief. Reserve its use for children with group A beta-hemolytic streptococcus pharyngitis who have moderate to severe pain, realizing that the benefit is of questionable significance. PMID- 12899808 TI - Do irbesartan and amlodipine reduce cardiovascular events in diabetic patients? AB - When added to antihypertensive treatment in patients with diabetes and nephropathy, neither the angiotensin receptor blocker (ARB) irbesartan nor the calcium channel blocker amlodipine reduced the overall occurrence of cardiovascular events. However, irbesartan decreased the rate of heart failure and amlodipine reduced the rate of acute myocardial infarction. PMID- 12899809 TI - Low-dose doxycycline moderately effective for acne. AB - The authors propose that moderate acne may be treated with doxycycline in subantimicrobial doses (20-mg tablets taken twice daily). This regimen was well tolerated, moderately effective in reducing skin lesions, and did not have a detectable effect on the antibiotic resistance of skin flora. The cost of Periostat (the only form of doxycycline 20 mg available in the US) is about 55 US dollars per month, while generic doxycycline 100 mg is about 10 US dollars. This study has some significant flaws, but a trial of low-dose doxycycline in an adult with acne severe enough to warrant antibiotics would still seem a reasonable, albeit expensive, option PMID- 12899810 TI - Selective aldosterone blockade reduces mortality after MI. AB - Eplerenone, a selective mineralocorticoid receptor antagonist currently approved by the Food and Drug Administration for treatment of hypertension, reduces mortality following myocardial infarction in patients with left ventricular dysfunction and clinical signs of congestive heart failure. Previous research by the same group established that spironolactone does the same at lower cost than eplerenone. PMID- 12899811 TI - Blood cultures not helpful for community-acquired pneumonia. AB - Blood cultures rarely contribute to the management of uncomplicated community acquired pneumonia. A positive blood culture has no relation to the severity of the illness or to patient outcomes. Physicians should order blood cultures only for those patients with severe illness and for those in whom initial therapy fails. PMID- 12899812 TI - Zonisamide effective for weight loss in women. AB - Zonisamide (Zonegran), in conjunction with a reduced-calorie diet (deficit of 500 kcal/d), resulted in an additional mean 5-kg (11-pound) weight loss compared with diet alone. This regimen was well-tolerated in obese female patients. Further evaluation of long-term side effects and continued weight loss beyond 32 weeks is needed. PMID- 12899813 TI - Continuous use of oral contraceptives reduces bleeding. AB - This study provides good evidence that continuous use of combination oral contraceptives for 1 year decreases bleeding without clinically important changes in blood pressure, weight, or hemoglobin when compared with cyclic users. Clinicians should consider offering this option to their patients, while continuing to look for evidence that addresses long-term sequelae, as well as patients of color or those with less than a college education. PMID- 12899814 TI - Clindamycin for vaginosis reduces prematurity and late miscarriage. AB - Using oral clindamycin to treat women with asymptomatic bacterial vaginosis during their second trimester (between 12 and 22 weeks estimated gestational age) reduces the number of premature births and late miscarriages. The study did not demonstrate a difference in the number of neonatal intensive care unit admissions, mean birth weight, or gestational age. This is the first study demonstrating benefits in treating asymptomatic bacterial vaginosis early in pregnancy. It still needs to be determined, in larger trials, whether generalized screening and treatment for asymptomatic bacterial vaginosis in the early second trimester is beneficial and cost-effective. PMID- 12899815 TI - Evaluation and management of hip pain: an algorithmic approach. AB - Start by determining whether pain is located in the anterior, lateral, or posterior hip. As the site varies, so does the etiology. Besides location, consider sudden vs insidious onset, motions and positions that reproduce pain, predisposing activities, and effect of ambulation or weight bearing. Physical examination tests that elucidate range of motion, muscle strength, and pain replication will narrow the diagnostic search. Magnetic resonance imaging is usually diagnostic if plain x-rays and conservative therapy are ineffective. Conservative measures and selective use of injection therapy are usually effective. PMID- 12899816 TI - ADHD treatment and academic performance: a case series. AB - Most new cases of attention deficit-hyperactivity disorder (ADHD) are of the predominantly inattentive subtype. Research on the use of psychostimulants in these patients has shown a high rate of nonresponders. Although psychostimulants showed a short-term decrease in symptoms in students diagnosed with predominantly inattentive ADHD, they did not significantly improve grade-point averages. PMID- 12899817 TI - Caribbean itch. PMID- 12899818 TI - Complementary medicine: where is the evidence? AB - Herbal medicines have been submitted to systematic reviews more frequently than any other complementary therapy, and it is here where the most positive evidence can be found. There is not much research into potential serious risks of complementary medicine. Possible risks range from the toxicity of herbs to vertebral artery dissection or nerve damage after chiropractic manipulation. Currently the Cochrane Library contains 34 systematic reviews of complementary medicine: 20 of herbal medicines, 7 of acupuncture, 3 of homeopathy, 2 of manual therapies, and 2 of other forms. PMID- 12899819 TI - Sibutramine in the management of obesity. PMID- 12899820 TI - Immediate- vs extended-release metoprolol in heart failure. PMID- 12899821 TI - Reducing emergency department visits among high-using patients. AB - Intervention using a real-time database system was accepted by physicians and reduced high-cost encounters. The risk of a high-cost encounter was significantly greater for the minimal intervention than for the moderate or maximal intervention groups. The probability of an emergency department visit was significantly reduced for minimal compared with moderate and maximal intervention. The risk for emergency department events was the same for the moderate and maximal intervention groups. Moderate intervention seems the most cost-effective because of reductions achieved with minimal staff involvement. PMID- 12899822 TI - Clinical inquiries. Heat or ice for acute ankle sprain? PMID- 12899823 TI - Clinical inquiries. How should patients with mitral regurgitation be followed? PMID- 12899824 TI - Clinical inquiries. Does glucosamine relieve arthritis joint pain? PMID- 12899825 TI - Clinical inquiries. Are nasal steroid sprays effective for otitis media with effusion? PMID- 12899826 TI - Transcriptional and post-transcriptional control of cold-shock genes. AB - A mesophile like Escherichia coli responds to abrupt temperature downshifts (e.g. from 37 degrees C to 10 degrees C) with an adaptive response that allows cell survival and eventually resumption of growth under the new unfavorable environmental conditions. During this response, bulk transcription and translation slow or come to an almost complete stop, while a set of about 26 cold shock genes is preferentially and transiently expressed. At least some of the proteins encoded by these genes are essential for survival in the cold, but none plays an exclusive role in cold adaptation, not even the "major cold-shock protein" CspA and none is induced de novo. The majority of these proteins binds nucleic acids and are involved in fundamental functions (DNA packaging, transcription, RNA degradation, translation, ribosome assembly, etc.). Although cold-induced activation of specific promoters has been implicated in upregulating some cold-shock genes, post-transcriptional mechanisms play a major role in cold adaptation; cold stress-induced changes of the RNA degradosome determine a drastic stabilization of the cold-shock transcripts and cold shock-induced modifications of the translational apparatus determine their preferential translation in the cold. This preferential translation at low temperature is due to cis elements present in the 5' untranslated region of at least some cold-shock mRNAs and to trans-acting factors whose levels are increased substantially by cold stress. Protein CspA and the three translation initiation factors (IF3 in particular), whose stoichiometry relative to the ribosomes is more than doubled during the acclimation period, are among the trans elements found to selectively stimulate cold-shock mRNA translation in the cold. PMID- 12899827 TI - Mapping the active sites of bacterial translation initiation factor IF3. AB - IF3C is the C-terminal domain of Escherichia coli translation initiation factor 3 (IF3) and is responsible for all functions of this translation initiation factor but for its ribosomal recycling. To map the number and nature of the active sites of IF3 and to identify the essential Arg residue(s) chemically modified with 2,3 butanedione, the eight arginine residues of IF3C were substituted by Lys, His, Ser and Leu, generating 32 variants that were tested in vitro for all known IF3 activities. The IF3-30S subunit interaction was inhibited strongly by substitutions of Arg99, Arg112, Arg116, Arg147 and Arg168, the positive charges being important at positions 116 and 147. The 70S ribosome dissociation was affected by mutations of Arg112, Arg147 and, to a lesser extent, of Arg99 and Arg116. Pseudo-initiation complex dissociation was impaired by substitution of Arg99 and Arg112 (whose positive charges are important) and, to a lesser extent, of Arg116, Arg129, Arg133 and Arg147, while the dissociation of non-canonical 30S initiation complexes was preserved at wild-type levels in all 32 mutants. Stimulation of mRNA translation was reduced by mutations of Arg116, Arg129 and, to a lesser extent, of Arg99, Arg112 and Arg131 whereas inhibition of non canonical mRNA translation was affected by substitutions of Arg99, Arg112, Arg168 and, to a lesser extent, Arg116, Arg129 and Arg131. Finally, repositioning the mRNA on the 30S subunit was affected weakly by mutations of Arg133, Arg131, Arg168, Arg147 and Arg129. Overall, the results define two active surfaces in IF3C, and indicate that the different functions of IF3 rely on different molecular mechanisms involving separate active sites. PMID- 12899828 TI - Positive and negative selection of mutant forms of prokaryotic (cyanobacterial) ribulose-1,5-bisphosphate carboxylase/oxygenase. AB - A system for biological selection of randomly mutagenized ribulose 1,5 bisphosphate carboxylase/oxygenase (Rubisco) genes from the cyanobacterium Synechococcus PCC6301 was designed in which a Rubisco deletion mutant of the photosynthetic bacterium Rhodobacter capsulatus served as a host. Trans complementation with the Synechococcus PCC6301 rbcLS genes enabled anaerobic photoautotrophic growth of the R.capsulatus deletion strain with 5% CO(2), but not with 1.5% CO(2) in the atmosphere, and this strain could not grow under aerobic chemoautotrophic conditions. Phenotypic differences between the R.capsulatus host strain complemented with the wild-type rbcLS genes and transconjugates carrying mutated genes were used to identify mutants that were able to complement to photoautotrophic growth with 1.5% CO(2). These "positive" mutant proteins were unaffected for any measured kinetic properties, with a single exception. A mutant with a valine substitution at phenylalanine 342 had an increased affinity for ribulose-1,5-bisphosphate. Mutants with changes in the affinity for CO(2) were isolated through negative selection, in which mutants incapable of complementing R.capsulatus to photoautotrophic growth with 5% CO(2) were identified. Mutations at aspartate 103 resulted in enzymes that were greatly affected for different kinetic parameters, including an increased K(m) for CO(2). This study demonstrated that random mutagenesis and bioselection procedures could be used to identify mutations that influence important properties of bacterial Rubisco; these residues would not have been identified by other methods. PMID- 12899829 TI - The frameshift stimulatory signal of human immunodeficiency virus type 1 group O is a pseudoknot. AB - Human immunodeficiency virus type 1 (HIV-1) requires a programmed -1 ribosomal frameshift to produce Gag-Pol, the precursor of its enzymatic activities. This frameshift occurs at a slippery sequence on the viral messenger RNA and is stimulated by a specific structure, downstream of the shift site. While in group M, the most abundant HIV-1 group, the frameshift stimulatory signal is an extended bulged stem-loop, we show here, using a combination of mutagenesis and probing studies, that it is a pseudoknot in group O. The mutagenesis and probing studies coupled to an in silico analysis show that group O pseudoknot is a hairpin-type pseudoknot with two coaxially stacked stems of eight base-pairs (stem 1 and stem 2), connected by single-stranded loops of 2nt (loop 1) and 20nt (loop 2). Mutations impairing formation of stem 1 or stem 2 of the pseudoknot reduce frameshift efficiency, whereas compensatory changes that allow re formation of these stems restore the frameshift efficiency to near wild-type level. The difference between the frameshift stimulatory signal of group O and group M supports the hypothesis that these groups originate from a different monkey to human transmission. PMID- 12899830 TI - Inverting enantioselectivity of Burkholderia cepacia KWI-56 lipase by combinatorial mutation and high-throughput screening using single-molecule PCR and in vitro expression. AB - The enantioselectivity of lipase from Burkhorderia cepacia KWI-56 has been inverted using a novel in vitro technique for construction and screening of a protein library by single-molecule DNA amplification by PCR followed by in vitro coupled transcription/translation system termed single-molecule-PCR-linked in vitro expression (SIMPLEX). Four amino acid residues (L17, F119, L167, and L266) in the hydrophobic substrate-binding pocket of the lipase were selected for mutation based on a structural model of a substrate-enzyme complex, and a combinatorial mutation library was constructed by SIMPLEX and screened for (R) and (S)-configurations of p-nitrophenyl 3-phenylbutyrate. Some combinations of amino acid substitutions in the four positions of the lipase were found as effective for changing the enantiopreference from the (S)-form substrate to the (R)-form. Two variants were expressed in the original host cells and purified to homogeneity, showing completely reversed enantioselectivity for the (R)-form of ethyl 3-phenylbutyrate (selectivity factor E(R)=38 or 33), whereas the wild-type lipase was (S)-selective (selectivity factor E(S)=33). Thus the semi-rational and semi-random combinatorial design of a mutant library followed by a high throughput screening based on their enzymatic activity should be a powerful tool to engineer the enantioselectivity of enzymes. PMID- 12899831 TI - Human 20alpha-hydroxysteroid dehydrogenase: crystallographic and site-directed mutagenesis studies lead to the identification of an alternative binding site for C21-steroids. AB - Human 20alpha-hydroxysteroid dehydrogenase (h20alpha-HSD; AKR1C1) catalyzes the transformation of progesterone (Prog) into 20alpha-hydroxy-progesterone (20alpha OHProg). Although h20alpha-HSD shares 98% sequence identity with human type 3 3alpha-HSD (h3alpha-HSD3, AKR1C2), these two enzymes differ greatly in their activities. In order to explain these differences, we have solved the crystal structure of h20alpha-HSD in a ternary complex with NADP(+) and 20alpha-OHProg at 1.59A resolution. The steroid is stabilized by numerous hydrophobic interactions and a hydrogen bond between its O20 and the N(epsilon ) atom of His222. This new interaction prevents the formation of a hydrogen bond with the cofactor, as seen in h3alpha-HSD3 ternary complexes. By combining structural, direct mutagenesis and kinetic studies, we found that the H(222)I substitution decreases the K(m) value for the cofactor 95-fold. With these results, we hypothesize that the rotation of the lateral chain of His222 could be a mediating step between the transformation of Prog and the release of the cofactor. Moreover, crystal structure analysis and direct mutagenesis experiments lead us to identify a new residue involved in the binding of Prog. Indeed, the R(304)L substitution leads to a 65-fold decrease in the K(m) value for Prog reduction. We thus propose that Prog is maintained in a new steroid-binding site composed mainly of residues found in the carboxy-terminal region of the protein. PMID- 12899832 TI - Sequence analysis of the genome of the temperate Yersinia enterocolitica phage PY54. AB - The temperate Yersinia phage PY54 belongs to the unusual group of phages that replicate as linear plasmids with covalently closed ends. Besides Escherichia coli phage N15, PY54 is the only member of this group to be identified. We have determined the complete sequence (46,339 bp) of the PY54 genome. Bioinformatic analyses revealed 67 open reading frames (ORFs) with good coding potential located on both DNA strands. The comparison of the deduced PY54 gene products with known proteins encoded by other phages and bacteria along with functional studies have enabled us to assign the possible functions of 25 ORFs. In the left arm of the PY54 genome, we identified a number of ORFs that obviously code for head and tail proteins. Furthermore, this part of the phage genome contains genes probably involved in plasmid partitioning. Regarding the predicted gene functions and gene order, the PY54 and N15 left arms are similar. However, there are only weak DNA homologies and, in contrast to N15, the Yersinia phage harbours only a few ORFs related to genes found in lambdoid phages. The PY54 right arm comprises mainly regulatory genes as well as genes important for plasmid replication, DNA methylation, and host cell lysis. Out of 36 deduced products of the right arm, 13 revealed strongest database homologies to N15 proteins, of which the protelomerase and the Rep protein are exclusively homologous to their N15 counterparts. A number of PY54 genes essential for the lytic or lysogenic cycle were identified by functional analysis and characterization of phage mutants. In order to study transcription during the lytic and lysogenic stage, we analysed 34 PY54 ORFs by reverse transcriptase (RT)-PCR. The phage transcription patterns in lysogenic bacteria and at the late lytic stage of infection are nearly identical. The reasons for this finding are spontaneous release of phages during lysogeny and a high rate of phages that lysogenize their Yersinia host upon infection. PMID- 12899833 TI - Sequence variability analysis of human class I and class II MHC molecules: functional and structural correlates of amino acid polymorphisms. AB - Major histocompatibility complex class I (MHCI) and class II (MHCII) molecules display peptides on antigen-presenting cell surfaces for subsequent T-cell recognition. Within the human population, allelic variation among the classical MHCI and II gene products is the basis for differential peptide binding, thymic repertoire bias and allograft rejection. While available 3D structural analysis suggests that polymorphisms are found primarily within the peptide-binding site, a broader informatic approach pinpointing functional polymorphisms relevant for immune recognition is currently lacking. To this end, we have now analyzed known human class I (774) and class II (485) alleles at each amino acid position using a variability metric (V). Polymorphisms (V>1) have been identified in residues that contact the peptide and/or T-cell receptor (TCR). Using sequence logos to investigate TCR contact sites on HLA molecules, we have identified conserved MHCI residues distinct from those of conserved MHCII residues. In addition, specific class II (HLA-DP, -DQ, -DR) and class I (HLA-A, -B, -C) contacts for TCR binding are revealed. We discuss these findings in the context of TCR restriction and alloreactivity. PMID- 12899834 TI - Crystal structure of alanine:glyoxylate aminotransferase and the relationship between genotype and enzymatic phenotype in primary hyperoxaluria type 1. AB - A deficiency of the liver-specific enzyme alanine:glyoxylate aminotransferase (AGT) is responsible for the potentially lethal hereditary kidney stone disease primary hyperoxaluria type 1 (PH1). Many of the mutations in the gene encoding AGT are associated with specific enzymatic phenotypes such as accelerated proteolysis (Ser205Pro), intra-peroxisomal aggregation (Gly41Arg), inhibition of pyridoxal phosphate binding and loss of catalytic activity (Gly82Glu), and peroxisome-to-mitochondrion mistargeting (Gly170Arg). Several mutations, including that responsible for AGT mistargeting, co-segregate and interact synergistically with a Pro11Leu polymorphism found at high frequency in the normal population. In order to gain further insights into the mechanistic link between genotype and enzymatic phenotype in PH1, we have determined the crystal structure of normal human AGT complexed to the competitive inhibitor amino oxyacetic acid to 2.5A. Analysis of this structure allows the effects of these mutations and polymorphism to be rationalised in terms of AGT tertiary and quaternary conformation, and in particular it provides a possible explanation for the Pro11Leu-Gly170Arg synergism that leads to AGT mistargeting. PMID- 12899835 TI - The crystal structure of Trypanosoma brucei enolase: visualisation of the inhibitory metal binding site III and potential as target for selective, irreversible inhibition. AB - The glycolytic enzymes of the trypanosomatids, that cause a variety of medically and agriculturally important diseases, are validated targets for drug design. Design of species-specific inhibitors is facilitated by the availability of structural data. Irreversible inhibitors, that bound covalently to the parasite enzyme alone, would be potentially particularly effective. Here we determine the crystal structure of enolase from Trypanosoma brucei and show that two cysteine residues, located in a water-filled cavity near the active-site, are modified by iodoacetamide leading to loss of catalytic activity. Since these residues are specific to the Trypanosomatidae lineage, this finding opens the way for the development of parasite-specific, irreversibly binding enolase inhibitors. In the present structure, the catalytic site is partially occupied by sulphate and two zinc ions. Surprisingly, one of these zinc ions illustrates the existence of a novel enolase-binding site for divalent metals. Evidence suggests that this is the first direct visualization of the elusive inhibitory metal site, whose existence has hitherto only been inferred from kinetic data. PMID- 12899836 TI - Strong bending of purple membranes in the M-state. AB - Structure changes of purple membranes during the photocycle were analysed in solution by measurements of the electric dichroism. The D96N-mutant was used to characterize the M-state at neutral pH. The transition from the resting state to 61% photo-stationary M-state is associated with a strong reduction of the dichroism decay time constant by a factor of approximately 2. Because the change of the time constant is independent of the bacteriorhodopsin concentration, the effect is not attributed to light-induced dissociation but to light-induced bending of purple membranes. After termination of light-activation the dichroism decay of the resting state is restored with a time constant close to that of the M-state decay, which is more than two orders of magnitude slower than proton transfer to the bulk. Thus, bending is not due to asymmetric protonation but to the structure of the M-state. A very similar reduction of decay time constants at a corresponding degree of light-activation was found for wild-type bacteriorhodopsin at pH-values 7.8-9.3, where the lifetime of the M-state is extended. Light-induced bending is also reflected in changes of the stationary dichroism, whereas the overall permanent dipole moment remains almost constant, suggesting compensation of changes in molecular and global contributions. Bead model simulations indicate that disks of approximately 1 microm diameter are bent at a degree of photo-activation of 61% to a radius of approximately 0.25 microm, assuming a cylindrical bending modus. The large light-induced bending effect is consistent with light-induced opening of the protein on the cytoplasmic side of the membrane detected by electron crystallography, which is amplified due to coupling of monomers in the membrane. Bending may function as a mechanical signal. PMID- 12899837 TI - Lipidic cubic phase crystal structure of the photosynthetic reaction centre from Rhodobacter sphaeroides at 2.35A resolution. AB - Well-ordered crystals of the bacterial photosynthetic reaction centre from Rhodobacter sphaeroides were grown from a lipidic cubic phase. Here, we report the type I crystal packing that results from this crystallisation medium, for which 3D crystals grow as stacked 2D crystals, and the reaction centre X-ray structure is refined to 2.35A resolution. In this crystal form, the location of the membrane bilayer could be assigned with confidence. A cardiolipin-binding site is found at the protein-protein interface within the membrane-spanning region, shedding light on the formation of crystal contacts within the membrane. A chloride-binding site was identified in the membrane-spanning region, which suggests a putative site for interaction with the light-harvesting complex I, the cytochrome bc(1) complex or PufX. Comparisons with the X-ray structures of this reaction centre deriving from detergent-based crystals are drawn, indicating that a slight compression occurs in this lipid-rich environment. PMID- 12899838 TI - Investigations into sequence and conformational dependence of backbone entropy, inter-basin dynamics and the Flory isolated-pair hypothesis for peptides. AB - The populations and transitions between Ramachandran basins are studied for combinations of the standard 20 amino acids in monomers, dimers and trimers using an implicit solvent Langevin dynamics algorithm and employing seven commonly used force-fields. Both the basin populations and inter-conversion rates are influenced by the nearest neighbor's conformation and identity, contrary to the Flory isolated-pair hypothesis. This conclusion is robust to the choice of force field, even though the use of different force-fields produces large variations in the populations and inter-conversion rates between the dominant helical, extended beta, and polyproline II basins. The computed variation of conformational and dynamical properties with different force-fields exceeds the difference between explicit and implicit solvent calculations using the same force-field. For all force-fields, the inter-basin transitions exhibit a directional dependence, with most transitions going through extended beta conformation, even when it is the least populated basin. The implications of these results are discussed in the context of estimates for the backbone entropy of single residues, and for the ability of all-atom simulations to reproduce experimental protein folding data. PMID- 12899839 TI - Structural evidence for the interaction of C-protein (MyBP-C) with actin and sequence identification of a possible actin-binding domain. AB - C-protein (MyBP-C) is a myosin-binding protein that is usually seen in two sets of seven to nine positions in the C-zones in each half of the vertebrate striated muscle A-band. Skeletal muscle C-protein is a modular structure containing ten sub-domains (C1 to C10) of which seven are immunoglobulin-type domains and three (C6, C7 and C9) are fibronectin-like domains. Cardiac muscle C-protein has an extra N-terminal domain (C0) and also some sequence insertions, one of which provides phosphorylation sites. It is conceivable that C-protein has both a structural and regulatory role within the sarcomere. The precise mode of binding of C-protein to the myosin filament has not been determined. However, detailed ultrastructural studies have suggested that C-protein, which binds to myosin, can give rise to a longer periodicity (about 435A) than the intrinsic myosin filament repeat of 429A. The reason for this has remained a puzzle for over 25 years. Here we show by modelling and computation that the presence of this longer periodicity could be explained if the myosin-binding part of C-protein binds to myosin with the expected 429A repeat, but if there are systematic interactions of the N terminal end of C-protein with the neighbouring actin filaments in the hexagonal lattice of filaments in the A-band. We also show that if they occur these interactions would probably only arise in defined muscle states. Further analysis of the MyBP-C sequence identifies a possible actin-binding domain in the Pro-Ala rich sequence found at the N terminus of skeletal MyBP-C and between domains C0 and C1 in the cardiac sequence. PMID- 12899840 TI - The molecular basis of vitamin E retention: structure of human alpha-tocopherol transfer protein. AB - Alpha-tocopherol transfer protein (alpha-TTP) is a liver protein responsible for the selective retention of alpha-tocopherol from dietary vitamin E, which is a mixture of alpha, beta, gamma, and delta-tocopherols and the corresponding tocotrienols. The alpha-TTP-mediated transfer of alpha-tocopherol into nascent VLDL is the major determinant of plasma alpha-tocopherol levels in humans. Mutations in the alpha-TTP gene have been detected in patients suffering from low plasma alpha-tocopherol and ataxia with isolated vitamin E deficiency (AVED). The crystal structure of alpha-TTP reveals two conformations. In its closed tocopherol-charged form, a mobile helical surface segment seals the hydrophobic binding pocket. In the presence of detergents, an open conformation is observed, which probably represents the membrane-bound form. The selectivity of alpha-TTP for RRR-alpha-tocopherol is explained from the van der Waals contacts occurring in the lipid-binding pocket. Mapping the known mutations leading to AVED onto the crystal structure shows that no mutations occur directly in the binding pocket. PMID- 12899841 TI - Changes in the expression of TGFbeta-isoforms in the anterior pituitary during withdrawal and resumption of feeding in hens. AB - Our goal was to determine whether TGFbeta-isoforms were involved in the remodeling of the pituitary cell population which occurred by regulation of feeding. The current study examined whether TGFbeta-isoforms were produced in the anterior pituitary, and the mRNA expression of TGFbeta-isoforms changed during withdrawal and resumption of feeding. White Leghorn laying hens were subjected to feed withdrawal for 4 days with a resumption of feeding thereafter. The anterior pituitary tissues were collected from hens of pretreatment (PT), 3 days after feed withdrawal (3DFW), 1 and 5 days after the resumption of feeding (1DRF and 5DRF, respectively), and on the day of resumption of egg-laying (RL). They were processed for semi-quantification of TGFbeta2, beta3, and beta4 mRNA expressions by reverse transcription-polymerase chain reaction (RT-PCR) and for immunocytochemistry for TGFbeta3. TGFbeta2, TGFbeta3, and TGFbeta4 mRNA expression with a product size of 269, 236, and 163bp, respectively, was observed in the anterior pituitaries in all groups of hens. Although the expression of TGFbeta2 and TGFbeta4 mRNA did not show any significant change, that of TGFbeta3 mRNA significantly declined in the 1DRF hens and recovered by the resumption of laying. Immunostaining revealed that TGFbeta3 was located in the cytoplasm of glandular cells with granule forms in all groups of hens. The ovarian and oviductal weights sharply declined in the 3DFW and 1DRF groups, followed by a full recovery in the RL hens. These results indicate that TGFbeta2, beta3, and beta4 were expressed in the chicken anterior pituitary, and TGFbeta3 mRNA expression was changed in correlation with the regulation of feeding, suggesting that this isoform may play a significant role in the regulation of the glandular cell population and/or differentiation. PMID- 12899842 TI - Profiles of PGH-alpha, GTH I-beta, and GTH II-beta mRNA transcript levels at different ovarian stages in the wild female Japanese eel Anguilla japonica. AB - The complete complementary DNA (cDNA) encoding pituitary gonadotropin II-beta subunit (GTH II-beta) of Japanese eel Anguilla japonica was cloned and sequenced, and the profiles of pituitary glycoprotein hormone alpha subunit (PGH-alpha), GTH I-beta, and GTH II-beta mRNA transcript levels at different stages of ovarian development before vitellogenesis in the wild females were investigated. The maturity of female eels was divided into four stages: juvenile, sub-adult, pre silver, and silver stages based on ovarian development and skin color. The GTH II beta cDNA was cloned by reverse transcription and polymerase chain reaction (RT PCR) amplification from total pituitary RNA. The full length GTH II-beta cDNA was obtained using 5(')- and 3(')-rapid amplification of cDNA ends. The cloned eel GTH II-beta cDNA consists of 646 bp nucleotides, including 53 bp nucleotides of 5(')-untranslated region (UTR), 423 bp of open reading frame, and 170 bp nucleotides of 3(')-UTR followed by a poly(A) tail. It encodes a 140-amino acid precursor molecule of GTH II-beta subunit with a putative signal peptide of 24 amino acids and a mature peptide of 116 amino acids. RT-PCR analysis showed that the pituitary transcript levels of alpha subunit steadily increased during eel silvering. The expression of GTH I-beta and II-beta mRNA levels, however, varied in different ovarian developmental stages. The mRNA expression of both GTH I-beta and GTH II-beta were detectable in juvenile stage. The expression levels of GTH II-beta mRNA, but not GTH I-beta, were significantly increased in sub-adult stage. The transcript levels of GTH I-beta and II-beta subunits further increased in pre-silver and silver stages. We demonstrated for the first time that the differential transcription patterns of pituitary PGH-alpha, GTH I-beta, and GTH II-beta mRNAs occur during silvering of the wild female Japanese eels. PMID- 12899843 TI - Environmental temperature increases plasma GH levels independently of nutritional status in rainbow trout (Oncorhynchus mykiss). AB - Like many poecilotherms, salmonids exhibit seasonal variations of growth rate in relation with seasonal temperatures and plasma GH level. However, temperature alters other parameters like food intake, which may directly modify the level of plasma GH. In order to determine whether temperature regulates plasma GH levels independently of nutritional status, fish were reared at 8, 12, or 16 degrees C and either fed ad libitum (fish with different food intake) to determine the global effect of temperature, or with the same ration (1.2%/body weight) to observe the temperature effect in fish with the same growth rate. Plasma insulin level was inversely proportional to the temperature (8, 12, and 16 degrees C) in fish fed ad libitum (12.1+/-0.3 ng/ml, 10.9+/-0.3 ng/ml, 9.5+/-0.4 ng/ml; P<0.001) and in restricted fish (14.0+/-0.3 ng/ml, 11.3+/-0.3 ng/ml, 10.0+/-0.2 ng/ml; P<0.0001), probably due to a prolonged nutrient absorption, and delayed recovery of basal insulin level at low temperature. Conversely, temperature did not affect plasma T3 level of fish fed ad libitum (2.5+/-0.2 ng/ml, 2.4+/-0.1 ng/ml, 2.5+/-0.1 ng/ml at 8, 12, and 16 degrees C) while fish fed with the same ration present less T3 at 16 degrees C than at 8 degrees C (1.83+/-0.1 ng/ml versus 1.2+/-0.1 ng/ml; P<0.001) throughout the experiment; these observations indicate that different plasma T3 levels reflect the different nutritional status of the fish. The levels of GH1 and GH2 mRNA, and GH1/GH2 ratio were not different for whatever the temperature or the nutritional status. Pituitary GH content, of fish fed ad libitum did not exhibit obvious differences at 8, 12, or 16 degrees C (254+/-9 ng/g bw, 237+/-18 ng/g bw, 236+/-18 ng/g bw), while fish fed with the same ration have higher pituitary GH contents at 16 degrees C than at 8 degrees C (401+/-30 ng/g bw versus 285+/-25 ng/g bw; P<0.0001). Interestingly, high temperature strongly increases plasma GH levels (2.5+/-0.3 ng/ml at 8 degrees C versus 4.8+/-0.6 ng/ml at 16 degrees C; P<0.0001) to the same extent in both experiments, since at a given temperature average plasma GH was similar between fish fed ad libitum or a restricted diet. Our results, demonstrate that temperature regulates plasma GH levels specifically but not pituitary GH content, nor the levels of GH1 and GH2 mRNA. In addition no differential regulation of both GH genes was evidenced whatever the temperature. PMID- 12899844 TI - Molecular characterization of the GnRH system in zebrafish (Danio rerio): cloning of chicken GnRH-II, adult brain expression patterns and pituitary content of salmon GnRH and chicken GnRH-II. AB - The zebrafish has proven to be a model system with unparalleled utility in vertebrate genetic and developmental studies. Substantially less attention has been paid to the potential role that zebrafish can play in answering important questions of vertebrate reproductive endocrinology. As an initial step towards exploiting the advantages that the zebrafish model offers, we have characterized their gonadotropin-releasing hormone (GnRH) system at the molecular level. GnRHs comprise a family of highly conserved decapeptide neurohormones widely recognized to orchestrate the hormonal control of reproduction in all vertebrates. We have isolated the gene and cDNA encoding chicken GnRH-II (cGnRH-II) from zebrafish, as well as several kilobases of upstream promoter sequence for this gene. As the gene encoding salmon GnRH (sGnRH) has been previously isolated (Torgersen et al, 2002), this is the second GnRH gene isolated from zebrafish to date. We have localized expression of these two genes in the brains of reproductively mature zebrafish using in situ hybridization. sGnRH is localized to the olfactory bulb terminal nerve region (OB-TN), the ventral telencephalon-preoptic area (VT-POA) and, as we report here for the first time in any teleost species, the hindbrain. cGnRH-II is expressed exclusively in the midbrain, as has been found in all other jawed vertebrate species examined. Finally, the levels of both GnRH peptides in pituitaries of reproductively mature zebrafish were quantified using specific ELISAs. sGnRH pituitary peptide levels were shown to be 3- to 4-fold higher than cGnRH-II pituitary peptide. The cumulative results of these experiments allow us to conclude that zebrafish express just two forms of GnRH in a site-specific manner within the brain, and that sGnRH is the hypophysiotropic GnRH form. This work lays the foundation for further research into the control of reproduction in zebrafish, such as the functional significance of multiple GnRHs in vertebrates, and the molecular mechanisms controlling tissue-specific GnRH expression. PMID- 12899845 TI - Estrogen receptors in Xenopus: duplicate genes, splice variants, and tissue specific expression. AB - The estrogenic steroid hormones, acting primarily through the nuclear estrogen receptors ERalpha and ERbeta, regulate sexual differentiation in a wide variety of vertebrates. In the frog Xenopus laevis, estrogen regulates the strength of vocal neuromuscular synapses and contributes to the physiological basis of sexually differentiated songs. To understand the mechanisms by which estrogen produces these effects, we have characterized the ERs of X. laevis and their expression in laryngeal muscle and other tissues. We found a remarkable molecular diversity in the estrogen receptor population within individuals. First, we have identified two distinct ERalpha genes, xlERalpha1 and xlERalpha2, which represent, to our knowledge, the first discovery of retained duplicates of the ERalpha gene in any species. These two genes are highly conserved at the amino acid level but have distinct nucleotide sequences; moreover, ERalpha2 has no N terminal domain. Cloning of ERalpha and ERbeta in the related species Xenopus tropicalis and phylogenetic analysis indicate that the two xlERalpha loci were generated by a duplication specific to the X. laevis lineage-most likely the genome duplication that led to a doubling of the X. laevis chromosome number about 30 million years ago. The primary ER expressed in X. laevis laryngeal muscle is the novel gene xlERalpha2; ERalpha1 is primarily expressed in liver, forebrain, and oviduct. Alternatively spliced transcripts of both xlERalpha1 and xlERalpha2 are also expressed in a tissue-specific manner. We propose that complementary spatial expression of these two genes and their alternatively spliced transcripts contributes to their conservation over such a long period of time, consistent with the subfunctionalization model for evolution after gene duplication. PMID- 12899846 TI - Distribution of adrenomedullin and proadrenomedullin N-terminal 20 peptide immunoreactivity in the pituitary gland of the frog Rana perezi. AB - Adrenomedullin (AM) and proadrenomedullin N-terminal 20 peptide (PAMP) are two multifunctional peptides processed from a common precursor which have been described in numerous mammalian organs, including the pituitary gland. Previous studies have found AM immunoreactivity in neurohypophysis nerve fibers of amphibian pituitary. In the present study, immunocytochemical and Western blot analysis in the pituitary gland of the amphibian Rana perezi demonstrated in the adenohypophysis both AM and PAMP. AM-like immunoreactivity was found in a moderate number of endocrine cells of the pars distalis. In the neurohypophysis, AM was observed not only in nerve fibers of pars nervosa and axonal projections innervating the pars intermedia, but also in the outer zone of the median eminence. PAMP staining was observed in numerous endocrine cells scattered all over the pars distalis and in some cells of the pars tuberalis, but not in the neurohypophysis. In order to compare the quantity of AM and PAMP immunoreactivity between pars distalis of female and male specimens, an image analysis study was done. Significant differences for AM immunoreactivity (p<0.001) between sexes was found, the males showing higher immunostained area percentage. Differences of PAMP immunoreactivity were not significant (p=0.599). Western blot analysis detected bands presumably corresponding to precursor and/or intermediate species in the propeptide processing. PMID- 12899847 TI - Dexamethasone induced preadipocyte recruitment and expression of CCAAT/enhancing binding protein alpha and peroxisome proliferator activated receptor-gamma proteins in porcine stromal-vascular (S-V) cell cultures obtained before and after the onset of fetal adipogenesis. AB - The present study examined the influence of dexamethasone (DEX) treatment on preadipocyte recruitment and expression of transcription factor proteins in adipose tissue stromal-vascular (S-V) cell cultures from 50 and 75 day old pig fetuses and young pigs. C/EBPalpha, C/EBPdelta, and PPARgamma immunoreactive cells had evenly reactive nuclei and unreactive nucleoli. DEX recruited many more preadipocytes in 75 day than in 50 day fetal S-V cultures. However, DEX did not increase the number of differentiated preadipocytes (lipid+, C/EBPalpha+) in 50 day S-V cultures and only slightly increased this number in 75 day fetal S-V cultures. In fetal cultures, extensive, precocious increases in C/EBPalpha expression (number of reactive cells) by day three were followed by extensive decreases in expression. However, PPARgamma expression was not expressed precociously since preadipocyte lipid accretion and PPARgamma immunoreactivity were strongly linked in fetal and pig S-V cultures. Nevertheless, all cells with lipid in fetal S-V cultures were C/EBPalpha and PPARgamma reactive. DEX increases preadipocyte differentiation in pig S-V cultures and in this study DEX increased PPARgamma expression to a much greater degree in pig than in fetal S-V cultures. These studies suggest that restricted adipogenesis in the pig fetus is attributable to limited DEX induced PPARgamma expression. PMID- 12899848 TI - Presence of the delta-MSH sequence in a proopiomelanocortin cDNA cloned from the pituitary of the galeoid shark, Heterodontus portusjacksoni. AB - Since a fourth MSH sequence, delta-MSH, has been detected in the proopiomelanocortin (POMC) gene of a dogfish and a stingray, members of superorder Squalea (class Chondrichthyes), it is possible that this novel MSH sequence might be a feature common to the POMC genes of all modern sharks and rays. As an initial step towards addressing this question, a full-length POMC cDNA was cloned and sequenced from the pituitary of the Port Jackson shark, Heterodontus portusjacksoni. The Port Jackson shark represents one of the oldest lineages in superorder Galea, and this superorder together with superorder Squalea form infraclass Neoselachii (the extant sharks and rays). The Port Jackson shark POMC cDNA has an open reading frame that is 1032 nucleotides in length and encodes the deduced amino acids sequences for beta-endorphin, ACTH/alpha-MSH, beta-MSH, gamma-MSH, and delta-MSH. Port Jackson shark delta-MSH has 83% primary sequence identity with dogfish and stingray delta-MSH, and it appears that the delta-MSH sequence may have been the result of an internal domain duplication and reinsertion of the beta-MSH sequence. The presence of the delta-MSH sequence in the POMC genes of representatives of both superorders of infraclass Neoselachii would indicate that the delta-MSH sequence must have been present in the ancestral euselachian shark that gave rise to the neoselachian radiation. PMID- 12899849 TI - Broiler chickens (Ross strain) lack insulin-responsive glucose transporter GLUT4 and have GLUT8 cDNA. AB - Identification of insulin-responsive glucose transporter proteins, GLUT4 and GLUT8, was attempted in chickens that characteristically are hyperglycemic and insulin resistant. Northern blot analysis using rat GLUT4 cDNA probe and RT-PCR using primers designed against the conserved regions in mammalian GLUT4 cDNA were not successful in identifying GLUT4 homologue(s) in various chicken tissues. Furthermore, GLUT4 homologues could not be detected in chicken tissues by genomic Southern blot analyses using a rat GLUT4 cDNA probe. These data, therefore, suggest that the GLUT4 homologous gene is deficient in chicken tissues. However, GLUT8, another insulin-responsive glucose transporter in the blastocyst, was identified with the aid of RACE (rapid amplification of cDNA ends) reactions in the chicken testis. Chicken GLUT8 was composed of 1449 bp with a coding region for a 482 amino acid protein. The deduced amino acid sequence was 58.8, 56.3, and 56.8% identical with human, rat, and mouse GLUT8, respectively. By RT-PCR, GLUT8 mRNA expressions were detected in chicken brain, kidney, adrenal, spleen, lung, testis, and pancreas; and barely detectable in skeletal muscle, liver, adipose tissue, and heart. Here we firstly report that GLUT8 was identified in chickens, while GLUT4, a major insulin-responsive transporter in mammals, is deficient in these animals. We propose the hypothesis that the hyperglycemia and insulin resistance observable in chickens is associated with their possible deficiency of GLUT4. PMID- 12899850 TI - Developmental expression and subcellular localization of wallaby gonadotropin releasing hormone receptor and its splice variants. AB - The developmental expression of gonadotropin-releasing hormone receptor (GnRH-R) and its splice variants was examined in the gonads of tammar wallaby pouch young in order to elucidate the functional role of GnRH-R in the developing testis and ovary. Wallaby GnRH-R, like eutherian GnRH-Rs, contains three exons and two introns. In the present study, the transcripts of two splice variants (GnRH-R Delta 1 and GnRH-R Delta 2) were cloned from the pituitary. GnRH-R Delta 1 contained a 291 bp deletion from nucleotide positions 232 to 522 within exon 1. This transcript appears to be distinctive in the wallaby and has not been reported in other species. GnRH-R Delta 2 contained a 220 bp deletion from nucleotide positions 523 to 742, corresponding to exon 2. We examined the subcellular localization of the wild type GnRH-R and its splice variants with confocal microscopy, showing that both the wild type receptor and the splice variants were membrane-associated molecules. The different pattern of expression of the wild type receptor and the variants transcripts found in adult and neonatal tissues suggests a specific developmental regulation of the GnRH-R Delta 2 transcript. In addition, the developmental expression of the GnRH-R and GnRH-R Delta 1 transcripts showed a possible association with key physiological events during gonadal development in the wallaby pouch young, suggesting that GnRH-R may be involved in the regulation of early development in the testis and ovary. PMID- 12899851 TI - In vitro progesterone production by maternal and embryonic tissues during gestation in the southern snow skink (Niveoscincus microlepidotus). AB - The southern snow skink, Niveoscincus microlepidotus, has a protracted gestation, lasting approximately one year. Ovulation occurs in spring (November) and embryonic development is completed by early autumn (March); however, birth does not occur until the following spring. Previous studies have shown that plasma progesterone concentration peaks in preovulatory females (spring, October), remains high during early gestation, and decreases to basal by autumn. In vitro progesterone production by corpora lutea, non-luteal ovary, anterior oviduct, placental tissues, muscle, and embryonic adrenal-gonads from N. microlepidotus was assessed throughout gestation. Tissues were incubated with or without the precursor pregnenolone for 3h at 24 degrees C; the resulting media were analysed for progesterone using radioimmunoassay. In vitro progesterone production by corpora lutea in media only was high during early gestation, dropping to basal by autumn. Maternal adrenal glands produced progesterone in vitro in media only throughout gestation; however, the pattern of production did not correlate with plasma concentrations and may represent steroid that is normally converted to corticosterone. Non-luteal ovary, anterior oviduct, placental tissues, muscle, and embryonic adrenal-gonads produced minimal progesterone in media only, but were able to convert pregnenolone to progesterone; this suggests steroid metabolic capability within these tissues. Further research is needed to address the possible endocrine role(s) of placental and embryonic tissues during gestation in viviparous squamates. PMID- 12899852 TI - Vitamin D metabolism in a frugivorous nocturnal mammal, the Egyptian fruit bat (Rousettus aegyptiacus). AB - The nocturnal, frugivorous Egyptian fruit bat (Rousettus aegyptiacus) has no obvious access to either endogenous or dietary sources of vitamin D. We hypothesized that this species under natural conditions would be vitamin D deficient and that both serum mineral concentrations and vitamin D metabolite concentrations would be low. Both wild populations and captive populations appear to have an impoverished vitamin D status, as concentrations of the principle circulating metabolite, 25-hydroxyvitamin D [25(OH)D] are undetectable (<4 ng/mL) and those of the active metabolite, 1,25-dihydroxyvitamin D [1,25(OH)(2)D] are low. Intraperitoneal administration of labelled 25(OH)D revealed enhanced 1 alpha hydroxylase activity confirming a natural state of vitamin D deficiency. This may account for the undetectable levels of 25(OH)D; for limited amounts of the prohormone substrate are rapidly converted to the active hormone. Both vitamin D(2) and D(3) metabolites were detected in bat serum, albeit in very small amounts, inferring that in their natural habitat fruit bats may have limited access to both exogenous dietary sources and endogenous sources. Despite the low levels of vitamin D metabolites in wild-caught and captive D-unsupplemented individuals, serum mineral concentrations were well regulated and similar to those of bats receiving D-supplements, with no pathological problems associated with vitamin D deficiency evident. PMID- 12899853 TI - Reproductive physiology of free-living White Ibises (Eudocimus albus) in the Florida Everglades. AB - We measured plasma concentrations of testosterone, estradiol, progesterone, and corticosterone; and recorded changes in gonad size, body condition, molt, and brood patch development of free-living adult White Ibises (Eudocimus albus) during the breeding season in the Florida Everglades. White Ibises are colonially breeding, long-legged wading birds that inhabit freshwater and estuarine wetlands. They have flexible breeding schedules (nest initiation dates can range from January to September) and onset of nesting is usually associated with increased prey availability caused by concentration of small fish in pools during periods of wetland drying. In this paper, we present the hormonal and physical characteristics of White Ibis reproductive physiology. We classified White Ibis breeding into five stages: pre-breeding, display, copulation/egg production, incubation, and chick rearing. White Ibises showed cyclic gonadal development which corresponded to reproductive stage. Male and female testosterone concentrations increased during the display stage and decreased during copulation, incubation, and chick rearing. Female estradiol concentrations were highest during display and chick rearing and male estradiol concentrations were lowest during copulation. Female progesterone concentrations increased during display and remained high throughout the breeding season. Female ibises had low corticosterone concentrations that increased during incubation and were highest during chick rearing, concomitant with lower body condition and flight muscle mass scores. Male ibis progesterone and corticosterone concentrations did not show seasonal changes and were more variable than concentrations in female ibises at similar stages. Males and females had elevated body condition scores during the display stage, which were depleted by the onset of incubation. Increased energy stores during display may be used later for fasting in male birds that do not eat during the 10-day copulation/egg production stage, and for egg production in female birds. During incubation, male and female ibises developed brood patches. Ibises molted in all stages of reproduction, indicating that ibis molt and reproductive physiology may not inhibit each other as in most temperate bird species. White Ibises showed similar patterns in reproductive physiology to other monogamous, seasonally breeding bird species in which both sexes incubate and care for the young. PMID- 12899854 TI - Vasopressin and galanin expression in the hypothalamus of two African rodents, Taterillus gracilis and Steatomys caurinus, subjected to water-restriction. AB - The expression of arginine-vasopressin (AVP) and galanin (GAL) was studied by immunohistochemistry and in situ hybridization in the hypothalamus of two species of African rodents. In the wild, these animals experience successive arid and wet seasons that alternately stimulate their antidiuretic and diuretic systems. In this study, animals were subjected to both standardized laboratory conditions and to eight days of water-restriction. Under both sets of conditions, AVP and GAL were detected in the supraoptic nucleus (SON), paraventricular nucleus (PVN), and median eminence (ME). AVP and GAL responses to water-restriction differed in the two species, as did behavioral adaptations to the hot-dry season. In Taterillus gracilis, AVP- and GAL-LI (like immunoreactivity) peptide and mRNA levels increased in the SON. AVP-LI peptide and mRNA levels increased in the PVN, whereas only AVP-LI peptide levels increased in the ME. Pituitary gland AVP pools were unchanged by water deprivation, whereas urinary AVP levels and osmolality increased. The AVP response is typical of that of desert rodents, favoring survival under conditions of water-restriction. In Steatomys caurinus, which estivates, AVP and GAL-LI peptide levels decreased in the hypothalamus, as they did in the laboratory rat. In the SON, AVP, and GAL mRNA levels increased, whereas, in the PVN, only AVP mRNA levels increased. Pituitary gland AVP levels decreased, whereas urinary AVP levels and osmolality increased. In both species, the changes in the amount of GAL-LI peptide appeared to be closely linked to changes in AVP levels, suggesting that this peptide is involved in the osmoregulatory response to water-restriction. PMID- 12899855 TI - Determination of tissue and plasma concentrations of PTHrP in fish: development and validation of a radioimmunoassay using a teleost 1-34 N-terminal peptide. AB - A specific and sensitive radioimmunoassay (RIA) for the N-terminus of sea bream (Sparus auratus) and flounder (Platichthys flesus) parathyroid hormone-related protein (PTHrP) was developed. A (1-34) amino-terminal sequence of flounder PTHrP was synthesized commercially and used as the antigen to generate specific antiserum. The same sequence with an added tyrosine (1-35(Tyr)) was used for iodination. Human (1-34) parathyroid hormone (PTH), human (1-34) PTHrP, and rat (1-34) PTHrP did not cross-react with the antiserum or displace the teleost peptide. Measurement of PTHrP in fish plasma was only possible after denaturing by heat treatment due to endogenous plasma binding activity. The minimum detectable concentration of (1-34) PTHrP in the assay was 2.5 pg/tube. The level of immunoreactive (1-34) PTHrP in plasma was 5.2+/-0.44 ng/ml (mean+/-SEM, n=20) for flounder and 2.5+/-0.29 ng/ml (n=64) for sea bream. Dilution curves of denatured fish plasma were parallel to the assay standard curve, indicating that the activity in the samples was indistinguishable immunologically from (1-34) PTHrP. Immunoreactivity was present, in order of abundance, in extracts of pituitary, oesophagus, kidney, head kidney, gills, intestine, skin, muscle, and liver. The pituitary gland and oesophagus contained the most abundant levels of PTHrP, 37.7+/-6.1 ng/g wet tissue and 2.3+/-0.7 ng/g wet tissue, respectively. The results suggest that in fish PTHrP may act in a paracrine and/or autocrine manner but may also be a classical hormone with the pituitary gland as a potential major source of the protein. PMID- 12899856 TI - Effects of sex steroids, sex, and sexual maturity on cortisol production: an in vitro comparison of chinook salmon and rainbow trout interrenals. AB - Sex steroids appear to be responsible for hyperactivation of the hypothalamus pituitary-interrenal (HPI) axis that occurs in mature semelparous Pacific salmon as a prelude to post-spawning (programmed) death. This study was undertaken to examine the direct effects of sex steroids on interrenal activity of semelparous (chinook salmon) and iteroparous (rainbow trout) salmonids using an in vitro incubation system. In addition, phenotypic sex differences in cortisol production by interrenals of sexually mature (spawning) rainbow trout and chinook salmon were investigated. Interrenal tissue from juvenile and sexually mature chinook salmon and rainbow trout was incubated for 48 h in culture medium containing either no steroid (controls), 1 microM estradiol (E2) or 1 microM 11 ketotestosterone (11-KT). This tissue was then challenged for 3h with either pregnenolone, dibutyryladenosine 3('):5(')-cyclic monophosphate (dbcAMP) or forskolin, or synthetic human adrenocorticotropic hormone (ACTH(1-24)). Sex differences in in vitro interrenal cortisol production were assessed using separate tissue pools challenged with the same agents. Cortisol in media was measured by radioimmunoassay. E2 suppressed the ability of juvenile chinook salmon interrenals to utilize pregnenolone as substrate for cortisol synthesis. In mature female chinook salmon the suppressive effect of E2 was less pronounced, but was observed as a reduced response of interrenals to both pregnenolone and dbcAMP. E2 did not affect ACTH(1-24) stimulated cortisol production. Immature and mature rainbow trout interrenals were both relatively insensitive to E2. 11-KT did not affect cortisol production by juvenile chinook salmon and juvenile or mature rainbow trout, and had only minor effects in male and female spawning chinook salmon. In mature chinook salmon and rainbow trout, the interrenals of females were more responsive to ACTH stimulation and showed a greater utilization of pregnenolone as a substrate than interrenals of males. Mature female rainbow trout were also more responsive to dbcAMP stimulation than males. The results of this study suggest that the onset of sexual maturation and gonadal steroid production may contribute to sexually dimorphic cortisol responses in vitro. PMID- 12899857 TI - Neurophysiologic correlates of psychiatric disorders and potential applications in epilepsy. AB - There is increasing interest in psychiatric assessment using neurophysiologic tools such as electroencephalography (EEG), magnetoencephalography (MEG), and transcranial magnetic stimulation (TMS). This is because these technologies have good temporal resolution, are relatively noninvasive, and (with the exception of MEG) are economical. Many different experimental paradigms and analysis techniques for the assessment of psychiatric patients involving these technologies are reviewed including conventional quantitative electroencephalography (QEEG), EEG cordance, low-resolution electromagnetic tomography (LORETA), frontal midline theta, midlatency auditory evoked potentials (P50, N100, P300), loudness dependency of the auditory evoked potential (LDAEP), mismatch negativity (MMN), contingent negative variation (CNV), and transcranial magnetic stimulation (TMS). Many of these neurophysiologic stimulus paradigms hold the promise of improving psychiatric patient care by improving diagnostic precision, predicting treatment response, and providing new phenotypes for genetic studies. Large cooperative multisite studies need to be designed to test and validate a few of these paradigms so that they might find use in routine clinical practice. PMID- 12899858 TI - Lamotrigine as adjunctive therapy in patients with refractory epilepsy and mental retardation. AB - This study evaluated the effects of lamotrigine as adjunctive therapy for refractory epilepsy in patients with mental retardation. Patients with epilepsy and mental retardation having uncontrolled seizures despite treatment with other antiepileptic drugs were eligible (n=67). The open-label study comprised a Baseline Phase, an Escalation Phase during which lamotrigine was titrated to a target dose, an 8-week Maintenance Phase during which doses of lamotrigine and concomitant antiepileptic drugs were maintained, and a 12-week Optimization Phase during which doses of lamotrigine and other antiepileptic drugs could be adjusted. Almost half (44%) of patients experienced a 50% reduction in seizure frequency during the Maintenance Phase after addition of lamotrigine; 15% of patients became seizure-free. A similar pattern of results was reported for the Optimization Phase. Investigator-rated clinical status was improved relative to baseline in 66 and 74% of patients at the end of the Maintenance and Optimization Phases, respectively. Most patients experienced improvements in seizure frequency, duration, and intensity during the Maintenance Phase (62 to 72%) and the Optimization Phase (65 to 74%). Many patients were rated as having improved social functioning during the Maintenance Phase (42%) and the Optimization Phase (46%). The Aberrant Behavior Checklist score for lethargy and the mean Habilitative Improvement Scale score were improved at the ends of the Maintenance and Optimization Phases relative to baseline (P< or =0.04). One limitation of this study is its open-label design, which limits the ability definitively to attribute the clinical improvements to lamotrigine. Adjunctive lamotrigine in patients with refractory epilepsy and mental retardation appears to decrease seizure frequency and improve behavior while permitting a reduction in dose of concomitant antiepileptic drugs. PMID- 12899859 TI - Sleep structure in patients with psychogenic nonepileptic seizures. AB - Psychogenic nonepileptic seizures (PNES) are a significant public health problem, occurring in perhaps 25% of patients admitted to epilepsy monitoring units. Additional distinguishing characteristics for these patients would be helpful from both a clinical and a scientific standpoint. This study examines sleep structure by polysomnography in patients with PNES compared with patients with epileptic seizures (ES). ES and PNES were verified by video-EEG monitoring. All patients with PNES were evaluated by a psychiatrist. Eight female patients with PNES were compared with 10 female ES patients in the same age group. Percentage REM sleep was significantly greater for women with PNES (23+/-1%) than for those with ES (18+/-1%). There were no differences in other sleep stages, total sleep time, or sleep efficiency. REM latency was less in PNES patients although not significantly. The study suggests that patients with PNES have a sleep architecture similar to that found in major depression, known to be associated with increased REM sleep. PMID- 12899860 TI - Mesial temporal lobe seizures may activate thermoregulatory mechanisms in humans: an infrared study of facial temperature. AB - Facial temperature T(f), an important link in cerebral thermoregulation and an ignored index of autonomic activity, was studied in three patients with mesial temporal epilepsy. Sets of infrared images were obtained automatically during seizures and also hourly, round-the-clock. For each patient, scan sets were sorted into: (1) ictal; (2) interictal with negligible motion; and (3) interictal with motion. Comparisons of mean, variance, and trend in each scan set among these groups showed considerable variability. However, significant trends in T(f) occurred (P<0.05) in two-thirds of seizures in one patient, three-fourths in another, and two-ninths in the third, consisting of steady decreases in T(f) beginning at peak seizure intensity R(max) and continuing into the postictus. Maximal ictal T(f) was greater than interictal T(f) (P<0.05), but it negatively correlated with R(max) in two subjects. The ictal changes in T(f), which may have a homeostatic role, suggest activation of thermoregulatory mechanisms during mesial temporal seizures. PMID- 12899861 TI - Relationship of sexual dysfunction to epilepsy laterality and reproductive hormone levels in women. AB - Sexual dysfunction has been reported to be common among women with epilepsy. Controlled studies, quantitative data, and investigations of potentially contributory factors, however, have been few. The purpose of this investigation was to determine if (1) sexual dysfunction is unusually common among women with partial seizures of temporal lobe origin (TLE), and (2) sexual dysfunction varies in relation to the laterality of EEG epileptiform discharges, antiepileptic drug use, and serum gonadal steroid levels. This controlled prospective investigation used a quantitative sexual rating scale and reproductive hormone measures to compare sexual dysfunction in women with left and right unilateral temporolimbic epilepsy and controls. Sexual dysfunction scores were significantly higher in women with TLE, and sexual dysfunction affected substantially more women with epilepsy than controls. Women with right-sided foci were affected more than women with left-sided foci. There was a significant inverse correlation between sexual dysfunction and bioactive testosterone levels in women with epilepsy as well as in controls. Serum estradiol was lower in women with TLE but did not correlate significantly with overall sexual dysfunction. The findings suggest that sexual dysfunction is significantly more common in women with right-sided epileptiform discharges than in controls and is inversely correlated with bioactive testosterone levels. The value of hormonal replacement or supplementation remains to be explored. PMID- 12899862 TI - Atypical hemispheric language dominance in left temporal lobe epilepsy as a result of the reorganization of language functions. AB - Hemispheric language dominance, as determined by intracarotid short-acting barbiturate injections (Wada testing), was retrospectively evaluated in 44 right handed patients with medically intractable left (n=26) or right (n=18) temporal lobe epilepsy (TLE). Atypical hemispheric language dominance (right or bilateral) was revealed in 13.6% of all patients investigated. A significantly higher rate of deviance from complete left hemisphere dominance was observed in left TLE (23.1%) than in right TLE (0%). Patient age at the time of seizure onset was proven a critical variable affecting atypical speech development in left TLE. The mean patient age at the time of seizure onset in left TLE patients with atypical language representation was 5.61 years, but was 13.13 years in patients with left hemisphere language dominance. Our results strongly support the influential impact of epileptiform activity spreading from the left temporal lobe structures on the reorganization of language functions in patients with early brain injury. PMID- 12899863 TI - The effect of lamotrigine on the EEGs of children and adolescents with epilepsy. AB - Lamotrigine (LTG) is one of the newer-generation antiepileptic drugs (AEDs) with broad-spectrum efficacy against a variety of seizures and epileptic syndromes. We retrospectively evaluated the effects of LTG as add-on therapy on EEGs of children and adolescents. The EEGs of 53 patients (mean age: 12.5 years) with primarily pharmacoresistant epilepsy were reviewed prior to and after LTG add-on therapy. Multiple seizure types were seen in 25, generalized seizures in 15, and complex partial seizures in 13 of the patients. Preceding LTG therapy, the baseline EEG was abnormal because of slow background in 60.3% and localized spikes in 35.8%, generalized spikes in 28.3%, or both in 24.5%. The EEG analysis during the 2-year follow-up period showed improvement in the background in 21.9%, interictal activity in 37.8%, and ictal pattern in 41.1% of the EEG recordings. Overall, LTG resulted in improvement in electrographic features which paralleled the clinical improvement. PMID- 12899864 TI - Psychogenic nonepileptic seizures: acute change in event frequency after presentation of the diagnosis. AB - Seizure frequency during inpatient video EEG monitoring was examined before and after the diagnosis of psychogenic nonepileptic seizures (PNES) was presented to patients (N=22). A control group of 10 patients with epileptic seizures (ES) were also followed from pre- to postdiagnosis. The number of PNES or ES within the 24 hour period prior to diagnosis was compared with the number of events that occurred within the 24-hour period after presentation of the diagnosis. Findings indicate that patients with PNES had a significant decrease in the frequency of events after diagnosis, while those with ES showed no change in event frequency after diagnosis. Eighteen of twenty-two patients with PNES had no further events during an acute follow-up period. Results suggest that providing patients with a diagnosis of PNES appears to reduce the acute frequency of PNES and may be an important first step in the long-term remediation of PNES. Long-term follow-up is needed to determine if such feedback alters the course of the disorder. PMID- 12899865 TI - The importance of being seizure free: topiramate and psychopathology in epilepsy. AB - The aim of this study was to analyze in detail psychopathology associated with topiramate (TPM) prescription and to analyze the relationship between psychopathology and seizure freedom. We analyzed the data on 103 patients who developed psychiatric disorders during TPM therapy. Forty-six patients developed an affective disorder, 22 aggressive behavior, 16 psychosis, 11 anxiety, and 8 personality changes such as anger, agitation, and hostile behavior. Patients with psychosis were more likely to be seizure-free during psychopathology, to receive psychotropic drug prescription, and to be admitted to hospital. In general, patients seizure-free during psychopathology were more likely to have a diagnosis of idiopathic generalized epilepsy, to be in co-therapy with vigabatrin, and to remit after drug reduction. We observed that psychopathology was related to seizure control in a subgroup of patients. The role of forced normalization, interlinked with interactions with other antiepileptic drugs, could be relevant. PMID- 12899866 TI - The influence of cognitive reserve on seizure-induced injury. AB - Status epilepticus (SE) is associated with a significant risk of cognitive impairment. While many factors likely determine cognitive outcome following SE, there is evidence that cognitive ability prior to a neurological insult may be an important determinant of outcome. Patients with greater cognitive abilities or so called cognitive reserve may be less vulnerable to injury than patients with limited cognitive ability. Here we tested the hypothesis that cognitive abilities prior to SE would be predictive of cognitive outcome. Immature rats were tested in the water maze, a test of visual-spatial memory, and divided into fast and slow learners. Animals were then subjected to SE and retested in the water maze 23 days later. Control rats were tested in the same manner but not subjected to SE. SE resulted in marked impairment in water maze performance. However, no statistical difference was noted in performance between slow and fast learners in either the SE or control group. Likewise, no differences were seen in the histopathology of the slow and fast learners. This study demonstrates that SE adversely effects visual-spatial memory equally in both fast and slow learners and does not support the theory that cerebral reserve plays a major role in cognitive function following a cerebral insult. PMID- 12899867 TI - Cognitive-behavioral group treatment program for adults with epilepsy in Hong Kong. AB - A subjective sense of uncertainty is a common theme in the experience of patients with epilepsy. Lack of control has also been implicated as an etiologic factor in the development of psychosocial problems in epilepsy. Cognitive-behavioral interventions emphasizing the mediating role of thoughts and self-control appear to be particularly relevant for patients with epilepsy because anxiety and depression are the most common problems. Despite its apparent importance, very little systematic research has been published on the effectiveness of psychological treatment of epilepsy. The purpose of the present study was to conduct a seminal outcome evaluation of psychological intervention for adult patients in Hong Kong. Using a waitlist control group, the treatment program focused particularly on cognitive restructuring and seizure control. Results indicated significant gains by the treatment group in overall quality of life and self-efficacy. In particular, marked improvements were noted for emotional well being, seizure, and stress management. The clinical implications are discussed in the context of developing specific intervention programs and outcome measures specific to the various needs of adult patients with epilepsy. PMID- 12899868 TI - Levetiracetam monotherapy during pregnancy: a case series. AB - The safety and efficacy of novel anticonvulsants during pregnancy in women with epilepsy are not well established. Although there are a variety of pregnancy registries accruing data on various outcomes, health care professionals managing women of childbearing age are interested in any information that may assist with clinical decisions. This case series summarizes the clinical outcomes of three women with epilepsy, all of whom took levetiracetam as monotherapy during pregnancy. PMID- 12899869 TI - Dramatic changes in artistic preference after left temporal lobectomy. PMID- 12899871 TI - Beyond LGMD1A: myotilin is a component of central core lesions and nemaline rods. AB - Myotilin is a Z-disc protein that binds alpha-actinin, gamma-filamin and F-actin. The essential role of myotilin in skeletal muscle is highlighted by the recent observation that autosomal dominant limb girdle muscular dystrophy type 1A is caused by mutations in the human myotilin gene. We studied the expression and subcellular distribution of myotilin in nemaline myopathy, central core disease, centronuclear myopathy, and myopathies with tubular aggregates. A prominent myotilin immunostaining of nemaline rods and core lesions was detected in all ten cases of nemaline myopathy and five cases of central core disease. This renders myotilin a sensitive, though non-specific marker for these structural lesions. Western blot analysis did not indicate an increased myotilin expression in nemaline myopathy muscle. However, the analysis indicated upregulation of a 75 kDa immunoreactive band, very weak in control muscle but previously detected in limb girdle muscular dystrophy 1A samples. Our findings indicate that myotilin is a core structural molecule in nemaline rods and central core lesions and suggest modification of myotilin in nemaline myopathy, and further support the notion that myotilin may have a key role in the dynamic molecular events mediating myofibril assembly in normal and diseased human skeletal muscle. PMID- 12899872 TI - Deficiency of the syntrophins and alpha-dystrobrevin in patients with inherited myopathy. AB - The syntrophins and dystrobrevins are members of the dystrophin-associated protein complex, and are thought to function as modular adaptors for signalling proteins recruited to the sarcolemmal membrane. We have characterised the expression of the syntrophins (alpha-, beta1-, and beta2-) and alpha-dystrobrevin by immunohistochemistry in normal human muscle and in biopsies from 162 patients with myopathies of unknown aetiology (with normal staining for dystrophin and other dystrophin-associated proteins). Unlike mice, beta2-syntrophin is expressed at the sarcolemma in post-natal human skeletal muscle. Deficiency of alpha dystrobrevin +/- beta2-syntrophin was present in 16/162 (10%) patients, compared to age-matched controls. All patients presented with congenital-onset hypotonia and weakness, although there was variability in clinical severity. Two major clinical patterns emerged: patients with deficiency of beta2-syntrophin and alpha dystrobrevin presented with severe congenital weakness and died in the first year of life, and two patients with deficiency of alpha-dystrobrevin had congenital muscular dystrophy with complete external ophthalmoplegia. We have sequenced the coding regions of alpha-dystrobrevin and beta2-syntrophin in these patients, and identified a new isoform of dystrobrevin, but have not identified any mutations. This suggests that disease causing mutations occur outside the coding region of these genes, in gene(s) encoding other components of the syntrophin-dystrobrevin subcomplex, or in gene(s) responsible for their post-translational modification and normal localisation. PMID- 12899873 TI - X-inactivation patterns in carriers of X-linked myotubular myopathy. AB - X-linked myotubular myopathy is a rare severe muscle disorder in affected male neonates. Most female carriers are free from symptoms. Skewed X inactivation has been proposed to be responsible for the affected phenotype seen in some carriers. We have compared the X inactivation patterns in blood DNA with the clinical phenotype in carriers of X-linked myotubular myopathy. The X-inactivation analysis was performed using HpaII predigestion of DNA followed by polymerase chain reaction of the highly polymorphic CAG repeat of the androgen receptor (AR) gene. The frequency of skewed X inactivation was similar in the X-linked myotubular myopathy carriers (22%) and in 235 controls (18%). Three overtly affected carriers had skewed X inactivation with the mutated X as the predominantly active X in at least two of them. Four females with mild symptoms had random X inactivation. The unaffected X-linked myotubular myopathy carriers had either skewed X inactivation in favour of expression from the normal X or random X-inactivation. Thus, there was a tendency for females with a more severe phenotype to have a skewed pattern of X inactivation, while females with an intermediate phenotype had a random pattern of X-inactivation. PMID- 12899874 TI - A Japanese family with FEOM1-linked congenital fibrosis of the extraocular muscles type 1 associated with spinal canal stenosis and refinement of the FEOM1 critical region. AB - We identified a Japanese family with congenital fibrosis of the extraocular muscles, including 24 affected individuals through five generations. To determine their form of congenital fibrosis of the extraocular muscles, we performed clinical and linkage studies. DNA typing for linkage to the FEOM1 (12p11.2-q12) and FEOM3 (16qter) loci was performed on genomic DNA, using fluorescent microsatellite polymorphic markers. All affected individuals shared the common manifestations of congenital fibrosis of the extraocular muscles type 1 including congenital ptosis, infraducted globe position in primary gaze, and upward gaze palsy in both eyes. Unexpectedly, we found apparent spinal canal stenosis in the cervical spine in all affected family members who were examined. Genetic analysis revealed linkage to the FEOM1 locus with a maximum lod score of 4.42 at theta of zero. One affected family member harbored a recombination event between D12S345 and D12S1692, narrowing the FEOM1 locus from the published 3-cM region flanked by D12S1584 and D12S1668 to a 2.1-cM region flanked by D12S345 and D12S1668. Thus, we have established that this family segregates congenital fibrosis of the extraocular muscles type 1 as an autosomal dominant trait and that their disorder both maps to and refines the FEOM1 locus. This is the first clinical and genetic report of such a family in the Japanese population and the first report of spinal involvement in congenital fibrosis of the extraocular muscles. PMID- 12899875 TI - Modification of the functional capacity of sarcoplasmic reticulum membranes in patients suffering from chronic fatigue syndrome. AB - In chronic fatigue syndrome, several reported alterations may be related to specific oxidative modifications in muscle. Since sarcoplasmic reticulum membranes are the basic structures involved in excitation-contraction coupling and the thiol groups of Ca(2+) channels of SR terminal cisternae are specific targets for reactive oxygen species, it is possible that excitation-contraction coupling is involved in this pathology. We investigated the possibility that abnormalities in this compartment are involved in the pathogenesis of chronic fatigue syndrome and consequently responsible for characteristic fatigue. The data presented here support this hypothesis and indicate that the sarcolemmal conduction system and some aspects of Ca(2+) transport are negatively influenced in chronic fatigue syndrome. In fact, both deregulation of pump activities (Na(+)/K(+) and Ca(2+)-ATPase) and alteration in the opening status of ryanodine channels may result from increased membrane fluidity involving sarcoplasmic reticulum membranes. PMID- 12899876 TI - Expression of plectin in muscle fibers with cytoarchitectural abnormalities. AB - Plectin is a protein belonging to the cytoskeletal anchoring system, concentrated at sites of mechanical stress in different cell types. In normal skeletal muscle, plectin is located at level of Z-discs, sarcolemma, post-synaptic membrane, and intermyofibrillar network. We investigated plectin immunocytochemistry in lobulated fibers, fibers with tubular aggregates, target fibers, central core disease and centronuclear myopathy. Thirty to forty percent of lobulated fibers had patchy increase of plectin immunoreactivity at sarcolemmal level with focal subsarcolemmal increases. Tubular aggregates revealed a low binding for plectin. Ten percent of central cores exhibited faint focal increase of plectin immunoreactivity. Target formations had a normal plectin pattern. In centronuclear myopathy, plectin immunoreactivity was increased around the centrally located nuclei in 8-12% of the fibers, at the sarcolemma of 50% of type 2 fibers, and at the membrane of small vacuoles located peripherally around the central nuclei. We postulate that plectin may play a role in the subsarcolemmal aggregation of mitochondria in the lobulated fibers, and in the central position of nuclei as well as in shape formation, positioning and moving of the vacuoles in centronuclear myopathy. PMID- 12899877 TI - The cranial sartorius muscle undergoes true hypertrophy in dogs with golden retriever muscular dystrophy. AB - The degree of atrophy or hypertrophy of selected pelvic limb muscles was determined in the canine homologue of Duchenne muscular dystrophy. While most muscles were atrophied, the caudal and cranial sartorius were hypertrophied. Cranial sartorius weights were corrected for body weight and endomysial space to determine true muscle weights (g/kg; mean+/-SD) in three golden retriever muscular dystrophy age groups, 4-10 (Group 1; n=15), 13-26 (Group 2; n=4), and 33 66 (Group 3; n=4) months and grouped normal dogs (6-20 months; n=12). Group 1 golden retriever muscular dystrophy weights (2.2063+/-0.6884) were greater than those of normal dogs (1.2699+/-0.1966), indicating that young golden retriever muscular dystrophy dogs have true cranial sartorius muscle hypertrophy. Values of Group 2 (1.3758+/-0.5078) and Group 3 (0.5720+/-0.2423) golden retriever muscular dystrophy dogs were less than those of Group 1, suggesting that the cranial sartorius muscle atrophies over time. Given that cranial sartorius muscle weight correlated with tarsal joint angle in affected dogs (r=-0.817), the hypertrophied muscle may play a role analogous to iliotibial band tightness in Duchenne muscular dystrophy. PMID- 12899878 TI - 109th ENMC International Workshop: 5th workshop on nemaline myopathy, 11th-13th October 2002, Naarden, The Netherlands. PMID- 12899879 TI - 108th ENMC International Workshop, 3rd Workshop of the MYO-CLUSTER project: EUROMEN, 7th International Emery-Dreifuss Muscular Dystrophy (EDMD) Workshop, 13 15 September 2002, Naarden, The Netherlands. PMID- 12899881 TI - Review of behaviour of oil in freezing environments. AB - The current knowledge of the physical fate and behaviour of crude oil and petroleum products spilled in Arctic situations is reviewed. The fate and final deposition of oil in marine conditions is presented as based on the extant literature. Spreading models were evaluated for oil on ice, under ice, in snow, in brash ice, and between blocks of ice. Models of oil transport under sheet and broken ice were considered, both for sea and river conditions. The ability of ice sheets to trap oil is discussed in relation to oil storage capacity. The effects of oil on a growing ice sheet were examined, both in terms of ice formation and the thermal effects of oil inclusions in ice. The migration of oil through ice was reviewed, focussing primarily on the movement through brine channels. The effects of oil on the surface of ice were considered, with emphasis on the effects of surface pools on ice melt. Similar consideration was given to the effects of oil on snow on the surface of ice. The few quantitative studies of oil in open and dynamic ice conditions are reviewed. Observations of intentional small-scale spills in leads and ice fields are reviewed and compared with observations from real spills. The conditions under which "oil pumping" from leads occurs were quantified. The most common ultimate fate of oil in an ice field is to be released onto the water surface. PMID- 12899882 TI - Development and application of damage assessment modeling: example assessment for the North Cape oil spill. AB - Natural resource damage assessment (NRDA) models for oil spills have been under development since 1984. Generally applicable (simplified) versions with built-in data sets are included in US government regulations for NRDAs in US waters. The most recent version of these models is SIMAP (Spill Impact Model Application Package), which contains oil fates and effects models that may be applied to any spill event and location in marine or freshwater environments. It is often not cost-effective or even possible to quantify spill impacts using field data collections. Modeling allows quantification of spill impacts using as much site specific data as available, either as input or as validation of model results. SIMAP was used for the North Cape oil spill in Rhode Island (USA) in January 1996, for injury quantification in the first and largest NRDA case to be performed under the 1996 Oil Pollution Act NRDA regulations. The case was successfully settled in 1999. This paper, which contains a description of the model and application to the North Cape spill, delineates and demonstrates the approach. PMID- 12899883 TI - Development and verification of deep-water blowout models. AB - Modeling of deep-water releases of gas and oil involves conventional plume theory in combination with thermodynamics and mass transfer calculations. The discharges can be understood in terms of multiphase plumes, where gas bubbles and oil droplets may separate from the water phase of the plume and rise to the surface independently. The gas may dissolve in the ambient water and/or form gas hydrates -a solid state of water resembling ice. All these processes will tend to deprive the plume as such of buoyancy, and in stratified water the plume rise will soon terminate. Slick formation will be governed by the surfacing of individual oil droplets in a depth and time variable current. This situation differs from the conditions observed during oil-and-gas blowouts in shallow and moderate water depths. In such cases, the bubble plume has been observed to rise to the surface and form a strong radial flow that contributes to a rapid spreading of the surfacing oil. The theories and behaviors involved in deepwater blowout cases are reviewed and compared to those for the shallow water blowout cases. PMID- 12899884 TI - Studies of the formation process of water-in-oil emulsions. AB - This paper summarizes studies to determine the formation process of water-in-oil emulsions and the stability of such emulsions formed in the laboratory and in a large test tank. These studies have confirmed that water-in-oil mixtures can be grouped into four states: stable emulsions, unstable water-in-oil mixtures, mesostable emulsions, and entrained water. These states are differentiated by rheological properties as well as by differences in visual appearance. The viscosity of a stable emulsion at a shear rate of one reciprocal second is about three orders of magnitude greater than that of the starting oil. An unstable emulsion usually has a viscosity no more than about 20 times greater than that of the starting oil. A stable emulsion has a significant elasticity, whereas an unstable emulsion does not. A mesostable emulsion has properties between stable and unstable, but breaks down within a few days of standing. The usual situation is that emulsions are either obviously stable, mesostable, or unstable. Entrained water, water suspended in oil by viscous forces alone, is also evident. Very few emulsions have questionable stability. Analytical techniques were developed to test these observations. The type of emulsion produced is determined primarily by the properties of the starting oil. The most important of these properties are the asphaltene and resin content and the viscosity of the oil. The composition and property ranges of the starting oil that would be required to form each of the water-in-oil states are discussed in this paper. PMID- 12899885 TI - Interaction of oil and mineral fines on shorelines: review and assessment. AB - The interaction of fine mineral particles with stranded oil in an aqueous medium reduces the adhesion of the oil to solid surfaces, such as sediments or bedrock. The net result is the formation of stable, micron-sized, oil droplets that disperse into the water column. In turn, the increase in surface area makes the oil more available for biodegradation. This interaction, referred to as oil mineral aggregate (OMA) formation, can explain how oiled shorelines are cleaned naturally in the absence of wave action in very sheltered coastal environments. OMA formation also plays an important role in the efficacy of shoreline treatment techniques, such as physical mixing and sediment relocation that move oiled sediments into the zone of wave action to promote the interaction between oil and mineral fines. Successful application of these shoreline treatment options has been demonstrated at two spill events (the Tampa Bay response in Florida and the Sea Empress operation in Wales) and at a controlled oil spill experiment in the field (the 1997 Svalbard ITOSS program). Sediment relocation harnesses the hydraulic action of waves so that the processes of fine-particle interaction and physical abrasion usually occur in tandem on open coasts. There has been no evidence of significant detrimental side-effects of residual oil in pelagic or benthic environments associated with the use of these treatment options to enhance rates of dispersion and oil biodegradation. PMID- 12899886 TI - Meso-scale testing and development of test procedures to maintain mass balance. AB - The Conrad Blucher Institute for Surveying and Science (Texas A&M University- Corpus Christi) has conducted numerous petroleum experiments at the Shoreline Environmental Research Facility (Corpus Christi, Texas, USA). The meso-scale facility has multiple wave tanks, permitting some control in experimental design of the investigations, but allowing for real-world conditions. This paper outlines the evolution of a materials balance approach in conducting petroleum experiments at the facility. The first attempt at a materials balance was during a 1998 study on the fate/effects of dispersant use on crude oil. Both water column and beach sediment samples were collected. For the materials balance, the defined environmental compartments for oil accumulation were sediments, water column, and the water surface, while the discharge from the tanks was presumed to be the primary sink. The "lessons learned" included a need to quantify oil adhesion to the tank surfaces. This was resolved by adhering strips of the polymer tank lining to the tank sides that could be later removed and extracted for oil. Also, a protocol was needed to quantify any floating oil on the water surface. A water surface (oil slick) quantification protocol was developed, involving the use of solid-phase extraction disks. This protocol was first tested during a shoreline cleaner experiment, and later refined in subsequent dispersant effectiveness studies. The effectiveness tests were designed to simulate shallow embayments which created the need for additional adjustments in the tanks. Since dispersant efficacy is largely affected by hydrodynamics, it was necessary to scale the hydrodynamic conditions of the tanks to those expected in our prototype system (Corpus Christi Bay, Texas). The use of a scaled model permits the experiment to be reproduced and/or evaluated under different conditions. To minimize wave reflection in the tank, a parabolic wave dissipater was built. In terms of materials balance, this design reduced available surface area as a sink for oil adsorption. PMID- 12899887 TI - The development of the SCAT process for the assessment of oiled shorelines. AB - The Shoreline Cleanup Assessment Team (SCAT) process is a tool to assess oiled shorelines and is now an integral component of spill response operations. The key element of a SCAT survey is a systematic documentation using standard terms and definitions of the shoreline in the areas affected by an oil spill. SCAT programs were initially established to provide objective and accurate shoreline oiling information directly to cleanup operations. The role of the SCAT program has since expanded and the information generated by the field teams is used now by planners and decision-makers and to develop shoreline treatment recommendations, to select appropriate treatment techniques, and to establish the level or degree of treatment that is appropriate. This latter point is an integral part of establishing shoreline treatment criteria or standards and treatment end points. PMID- 12899888 TI - Development of oil hydrocarbon fingerprinting and identification techniques. AB - Oil, refined product, and pyrogenic hydrocarbons are the most frequently discovered contaminants in the environment. To effectively determine the fate of spilled oil in the environment and to successfully identify source(s) of spilled oil and petroleum products is, therefore, extremely important in many oil-related environmental studies and liability cases. This article briefly reviews the recent development of chemical analysis methodologies which are most frequently used in oil spill characterization and identification studies and environmental forensic investigations. The fingerprinting and data interpretation techniques discussed include oil spill identification protocol, tiered analytical approach, generic features and chemical composition of oils, effects of weathering on hydrocarbon fingerprinting, recognition of distribution patterns of petroleum hydrocarbons, oil type screening and differentiation, analysis of "source specific marker" compounds, determination of diagnostic ratios of specific oil constituents, stable isotopic analysis, application of various statistical and numerical analysis tools, and application of other analytical techniques. The issue of how biogenic and pyrogenic hydrocarbons are distinguished from petrogenic hydrocarbons is also addressed. PMID- 12899889 TI - MORICE--new technology for mechanical oil recovery in ice infested waters. AB - Mechanical oil recovery in ice infested waters (MORICE) was initiated in 1995 to develop technology for the recovery of oil spills in ice. It has been a multinational effort involving Norwegian, Canadian, American and German organizations and researchers. Through a stepwise approach with the development organized in six separate phases, laboratory tests and field experiments have been conducted to study various ideas and concepts, and to refine the ideas that were considered to have the best potential for removing oil in ice. Put together in one unit, these concepts included ice processing equipment and two alternative oil recovery units installed on a work platform. In January 2002, the final oil and ice testing with MORICE concepts was conducted at the Ohmsett test facility in Leonardo, New Jersey. The unit has been referred to as a harbor version to indicate the size and operating conditions, but the concepts could be scaled up to increase the capacity of oil and ice processing. For heavier ice conditions it would also be necessary to increase the overall strength. PMID- 12899890 TI - Latest update of tests and improvements to US Coast Guard viscous oil pumping system. AB - Over the past nine years, the US Coast Guard has incorporated the prevention through people (PTP) philosophy as a "human factors" approach to learn how maritime operations can be regulated safer and be more efficient by evaluating training, management policies, operational procedures, and establishing partnerships with the maritime industry. One of the key elements of applying a PTP approach is identifying and incorporating lessons learned from major marine casualties and pollution incidents. Since 1997, the US Coast Guard National Strike Force has responded to three major oil spills involving foreign freight vessels grounding, which included the removal of highly viscous oil using various lightering equipment and systems. An informal workgroup consisting of the US Coast Guard, US Navy Supervisor of Salvage (NAVSUPSALV), and various representatives from oil pollution clean-up companies met at the following facilities: the Chevron Asphalt Facility in Edmonds, WA (September 1999), the Oil and Hazardous Materials Simulated Environmental Test Tank (OHMSETT) testing facility in Leonardo, New Jersey (November 1999 and March 2000), the Alaska Clean Seas (ACS) warehouse annex in Prudhoe Bay, AK (October 2000), and Cenac Towing Company facility in Houma, LA (May 2002). The group shared ideas and techniques, and tested different pumps and hose lengths with viscous oil. It was during the early tests that the first quantitative results showed just how efficient lubricated transport of heavy oil product could be, and broadened the knowledge of such methods to the entire industry. Although this technology had existed for many years in the oil production and handling industry, its use had never been investigated in a laboratory setting with regard to salvage response lightering systems. PMID- 12899891 TI - Review of the development of laser fluorosensors for oil spill application. AB - As laser fluorosensors provide their own source of excitation, they are known as active sensors. Being active sensors, laser fluorosensors can be employed around the clock, in daylight or in total darkness. Certain compounds, such as aromatic hydrocarbons, present in petroleum oils absorb ultraviolet laser light and become electronically excited. This excitation is quickly removed by the process of fluorescence emission, primarily in the visible region of the spectrum. By careful choice of the excitation laser wavelength and range-gated detection at selected emission wavelengths, petroleum oils can be detected and classified into three broad categories: light refined, crude or heavy refined. This paper will review the development of laser fluorosensors for oil spill application, with emphasis on system components such as excitation laser source, and detection schemes that allow these unique sensors to be employed for the detection and classification of petroleum oils. There have been a number of laser fluorosensors developed in recent years, many of which are strictly research and development tools. Certain of these fluorosensors have been ship-borne instruments that have been mounted in aircraft for the occasional airborne mission. Other systems are mounted permanently on aircraft for use in either surveillance or spill response roles. PMID- 12899892 TI - Development of airborne oil thickness measurements. AB - A laboratory sensor has now been developed to measure the absolute thickness of oil on water slicks. This prototype oil slick thickness measurement system is known as the laser-ultrasonic remote sensing of oil thickness (LURSOT) sensor. This laser opto-acoustic sensor is the initial step in the ultimate goal of providing an airborne sensor with the ability to remotely measure oil-on-water slick thickness. The LURSOT sensor employs three lasers to produce and measure the time-of-flight of ultrasonic waves in oil and hence provide a direct measurement of oil slick thickness. The successful application of this technology to the measurement of oil slick thickness will benefit the scientific community as a whole by providing information about the dynamics of oil slick spreading and the spill responder by providing a measurement of the effectiveness of spill countermeasures such as dispersant application and in situ burning. This paper will provide a review of early developments and discuss the current state-of-the art in the field of oil slick thickness measurement. PMID- 12899893 TI - Impact of training on rating competence in cognitive therapy. AB - This naturalistic study examines the impact of a training package on the inter rater reliability scores of a scale devised to measure competence in cognitive therapy (CT). The study was conducted at the Newcastle Cognitive and Behavioural Therapies Centre, where trainees are routinely required to submit videotapes of their therapeutic work for marking independently by two supervisors. This process was used as vehicle for the present work, whereby the trainees' videos were assessed in terms of their competence using a recently revised rating scale (CTS R). The inter-rater reliability of the supervisors' ratings was assessed both before and after the administration of a teaching package designed to improve the reliability in the use of the CTS-R. The results showed an improvement in the inter-rater agreement of five out of 12 items of the scale. The overall agreement also improved significantly following training. PMID- 12899894 TI - Multiple-cue probabilistic learning in spider-fearful and in panic-prone individuals. AB - Probabilistic learning was studied in spider-fearful and in panic-prone individuals. Participants learned to predict the occurrence of aversive outcomes on the basis of relevant and irrelevant stimuli. The salience of irrelevant stimuli was manipulated by the use of threat-related objects, i.e. pictures of spiders and pictures of emergency situations. In spider-fearful individuals, the presence of irrelevant pictures of spiders interfered with the adequate acquisition of the association between neutral relevant stimuli and aversive outcomes. Salient, i.e. threat-related, irrelevant stimuli exerted competitive effects on the accruing of associative strengths between neutral stimuli and aversive outcomes in spider-fearful, but not in panic-prone, individuals. PMID- 12899895 TI - Differential aversive outcome expectancies for high- and low-predation fear relevant animals. AB - There is now considerable evidence that phobic responding is associated with a bias towards expecting aversive or traumatic outcomes following encounters with the phobic stimulus (e.g. Behavioural Brain Sci. 18 (1995) 289-325; Phobias: A Handbook of Theory, Research and Treatment. Wiley, Chichester, 1997). In terms of conditioning contingencies, this can be described as a bias towards expecting an aversive unconditioned stimulus (UCS) following a phobic conditioned stimulus (CS). The disease-avoidance model of animal fears (Anxiety Res. 4 (1992a) 314; Matchett and Davey, 1991) suggests that common animal fears may be mediated by at least two kinds of selective associations: (1) a bias towards expecting physically harmful consequences associated with predatory animals, and (2) a bias towards expecting disgust or disease-relevant consequences associated with animals that are fear-relevant (FR) but normally physically harmless. The present study investigated this model of selective associations by comparing the UCS expectations elicited by high-predation FR, low-predation FR and safe (fear irrelevant) animals. The results indicate that high-predation animals are selectively associated with a pain relevant UCS, whilst low-predation animals are selectively associated with a disgust-relevant UCS. Safe animals were not strongly associated with either class of UCS. These findings provide evidence for a possible associative mechanism by which changes in nonspecific levels of disgust sensitivity may directly affect levels of fear to low-predation FR animals. PMID- 12899896 TI - Treatment of panic disorder via the Internet: a randomized trial of CBT vs. applied relaxation. AB - A randomized trial was conducted of two different self-help programs for panic disorder (PD) on the Internet. After confirming the PD-diagnosis with an in person structured clinical interview for DSM-IV (SCID) interview 22 participants were randomized to either applied relaxation (AR) or a multimodal treatment package based on cognitive behavioral therapy (CBT). Overall, the results suggest that Internet-administered self-help plus minimal therapist contact via e-mail has a significant medium to large effect (Cohen's d=0.71 for AR and d=0.42 for CBT). The results from this study generally provide evidence to support the continued use and development of Internet-distributed self-help programs. PMID- 12899897 TI - The perseveration of checking thoughts and mood-as-input hypothesis. AB - This paper describes two experiments designed to investigate how a current model of task perseveration, the mood-as-input hypothesis, might be applied to activities relevant to compulsive checking. The mood-as-input hypothesis predicts that perseveration at an open-ended task will be determined by a combination of the "stop rules" adopted for the task, and the valency of the mood state in which the task is conducted. Experiment 1 required participants to generate items that should be checked for safety/security if they were leaving their home unattended. Experiment 2 used an analogue recall task, in which participants were asked to recall items from a comprehensive list of items that should be checked if they were to leave their home safe/secure. Both experiments found that perseveration at the tasks was determined by particular configurations of mood and stop rules for the task. Of most relevance to compulsive checking was the fact that facilitated perseveration occurred when participants were asked to undertake the tasks in a negative mood using "as many as can" stop rules. Implications for the factors that develop and maintain compulsive checking are discussed. PMID- 12899898 TI - Emotional responding to hyperventilation as a predictor of agoraphobia status among individuals suffering from panic disorder. AB - Some data suggest that panic patients with extensive agoraphobia (PDA) display more intense respiratory distress during their panic attacks than Panic disorder (PD) patients. However, no studies have determined if PDA patients also show heightened sensitivity to a respiratory challenge compared to PD patients. The current study examined the differential emotional responding to hyperventilation among PDA patients, PD patients, and a non-clinical group with a history of panic attacks. Response to hyperventilation challenge did not distinguish non-clinical panickers from panic patients; however, behavioral tolerance to hyperventilation challenge significantly predicted agoraphobia status among panic disorder patients, even after controlling for demographic and clinical status variables. PMID- 12899899 TI - Personality pathology and cognitive-behavioral treatment of fear of flying. AB - Studies have been inconclusive about the influence of personality pathology on treatment outcome in anxiety disorders. In general, it has been presumed that treatment outcome is negatively influenced by the presence of personality pathology. This is a study of the prevalence of personality pathology among persons who were seeking help for fear of flying. Moreover, the effects of personality pathology on the results of a multimodal, standardized, cognitive behavioral fear of flying treatment program employed by an agency that specializes in treating people with fear of flying were studied. Personality pathology was determined with a self-report questionnaire, which provides ICD-10 diagnoses of personality disorders and dimensional severity scores for personality pathology. Treatment outcome was assessed with three different fear of flying questionnaires. Based on clinical judgment after individual-case conceptualization, participants (N=922) were assigned to a particular treatment for fear of flying. Self-report data for fear of flying were collected at pretreatment and at 3, 6 and 12-month follow-ups in 659 participants who followed the 2-day treatment program. Moreover, the number of flights made in the year following treatment was determined.The results of this study showed that participants with personality pathology, mainly from cluster C (anxiety), report greater fear of flying before treatment than participants without personality pathology. After treatment fear of flying was significantly reduced. Presence of personality pathology was not predictive of the number of flights after treatment and scores on the VAFAS scale at short or long term. Only on two questionnaires for fear of flying collected at short-term participants with personality pathology obtained significantly higher scores, although the size of the differences was relatively small. It was concluded that participants with personality pathology also benefited from fear of flying treatment and that the presence of personality pathology although cannot be regarded as a contra indication for a standardized, cognitive-behavioral group treatment. PMID- 12899900 TI - Methods for studying private sector supply of public health products in developing countries: a conceptual framework and review. AB - The private sector is an important supplier of public health products (PHPs) in developing countries. Although there are concerns about the quality and affordability of these products, private providers also offer possibilities for expanding access to key commodities. This paper proposes a conceptual framework for understanding the public health implications of private sales of PHPs. It reviews methods for studying these sales, together with their advantages and shortcomings. Ten methods are identified which can be used for studying the behaviour of providers and consumers. The effects of seasonal variation are discussed, together with the challenges of creating a sampling frame and studying illicit behaviour. We conclude that relatively little is known about the sales of PHPs, that more is known about contraceptives and drugs than about the newer products, and that the demand side of the market has been studied in greater depth than the behaviour of suppliers. The existing toolbox is biased towards formal providers, and thus, probably towards understanding the provision of PHPs to those who are better off. Methods for studying the supply of PHPs in outlets used by poor people is a priority area for further methodological development. PMID- 12899901 TI - The moral relevance of personal characteristics in setting health care priorities. AB - This paper discusses the moral relevance of accounting for various personal characteristics when prioritising between groups of patients. After a review of the results from empirical studies, we discuss the ethical reasons which might explain-and justify-the views expressed in these studies. The paper develops a general framework based upon the causes of ill health and the consequences of treatment. It then turns to the question of the extent to which a personal characteristic-and the eventual underlying ethical justification of its relevance could have any relationships to these causes and consequences. We attempt to disentangle those characteristics that may reflect a potentially relevant justification from those which violate widely accepted principles of social justice. PMID- 12899902 TI - Are seasonalities in suicide dependent on suicide methods? A reappraisal. AB - In general, suicide frequencies peak in spring and early summer and depict a low in autumn and winter months. Suicide seasonality is a ubiquitous phenomenon; however, recent studies have indicated that it is related particularly to violent suicide methods. We investigated differences in suicide seasonalities by suicide methods using Swiss mortality data. The suicide data were extracted from the Swiss cause of death statistics and cover the period 1969-1994. In this period 37,518 suicides (ICD-8 codes 950-959) were registered in the mortality statistics. The data were analysed by the Edwards' method, which examines unimodal sinusoidal patterns in aggregated monthly data, and by spectral analysis with disaggregated monthly data. Overall, Swiss suicide data in 1969-1994 depict the expected pattern with a peak in May and June and a low in December. However, analyses by suicide methods show that there is no common pattern. Suicide seasonalities are absent or weak in some violent as well as non-violent methods (firearms, cutting, crashing, poisoning). Even if clearly present (hanging, drowning, jumping from high places, unspecified/other), the seasonalities show distinct patterns and cycles. Apart from sinusoidal patterns, there are also outliers: in overall data as well as in most method-specific data, the December frequencies fall more than 10% below the neighbouring months. The decrease of the frequencies culminates at Christmas/New Year and then returns to average levels. Hence, the seasonality of the overall Swiss suicide frequencies is but a compound seasonality. It largely depends on specific suicide methods and different cyclical dynamics. PMID- 12899903 TI - Urban-rural differences in suicide trends in young adults: England and Wales, 1981-1998. AB - Suicide rates amongst young people, particularly males, have increased in many industrialised countries since the 1960s. There is evidence from some countries that the steepest rises have occurred in rural areas. We have investigated whether similar geographical differences in trends in suicide exist in England and Wales by examining patterns of suicide between 1981 and 1998 in relation to rurality. We used two complementary population-based indices of rurality: (1) population density and (2) population potential (a measure of geographic remoteness from large concentrations of population). We used the electoral ward (n=9264, median population aged 15-44: 1829) as the unit of analysis. To assess whether social and economic factors underlie rural-urban differences in trends we used negative binomial regression models to investigate changes in suicide rates between the years for which detailed national census data were available (1981 and 1991). Over the years studied, the most unfavourable trends in suicide in 15 44-year olds generally occurred in areas remote from the main centres of population; this effect was most marked in 15-24-year-old females. Observed patterns were not explained by changes in age- and sex-specific unemployment, socio-economic deprivation or social fragmentation. The mental health of young adults or other factors influencing suicide risk may have deteriorated more in rural than urban areas in recent years. Explanations for these trends require further investigation. PMID- 12899904 TI - Social capital and trust in providers. AB - Trust in providers has been in decline in recent decades. This study attempts to identify sources of trust in characteristics of health care systems and the wider community. The design is cross-sectional. Data are from (1) the 1996 Household Survey of the Community Tracking Study, drawn from 24 Metropolitan Statistical Areas; (2) a 1996 multi-city broadcast media marketing database including key social capital indicators; (3) Interstudy; (4) the American Hospital Association; and (5) the American Medical Association. Independent variables include individual socio-demographic variables, HMO enrollment, community-level health sector variables, and social capital. The dependent variable is self-reported trust in physicians. Data are merged from the various sources and analyzed using SUDAAN. Subjects include adults in the Household Survey who responded to the items on trust in physicians (N=17,653). Trust in physicians is independently predicted by community social capital (p<0.001). Trust is also negatively related to HMO enrollment and to many individual characteristics. The effect of HMOs is not uniform across all communities. Social capital plays a role in how health care is perceived by citizens, and how health care is delivered by providers. Efforts to build trust and collaboration in a community may improve trust in physicians, health care quality, access, and preserve local health care control. PMID- 12899905 TI - Declining subscriptions to the Maliando Mutual Health Organisation in Guinea Conakry (West Africa): what is going wrong? AB - Mutual Health Organisations (MHOs) are a type of community health insurance scheme that are being developed and promoted in sub-Saharan Africa. In 1998, an MHO was organised in a rural district of Guinea to improve access to quality health care. Households paid an annual insurance fee of about US$2 per individual. Contributions were voluntary. The benefit package included free access to all first line health care services (except for a small co-payment), free paediatric care, free emergency surgical care and free obstetric care at the district hospital. Also included were part of the cost of emergency transport to the hospital. In 1998, the MHO covered 8% of the target population, but, by 1999, the subscription rate had dropped to about 6%. In March 2000, focus groups were held with members and non-members of the scheme to find out why subscription rates were so low. The research indicated that a failure to understand the scheme does not explain these low rates. On the contrary, the great majority of research subjects, members and non-members alike, acquired a very accurate understanding of the concepts and principles underlying health insurance. They value the system's re-distributive effects, which goes beyond household, next of kin or village. The participants accurately point out the sharp differences that exist between traditional financial mechanisms and the principle of health insurance, as well as the advantages and disadvantages of both. The ease with which risk pooling is accepted as a financial mechanism which addresses specific needs demonstrates that it is not, per se, necessary to build health insurance schemes on existing or traditional systems of mutual aid. The majority of the participants consider the individual premium of 2 US dollars to be fair. There is, however, a problem of affordability for many poor and/or large families who cannot raise enough money to pay the subscription for all household members in one go. However, the main reason for the lack of interest in the scheme, is the poor quality of care offered to members of the MHO at the health centre. PMID- 12899906 TI - Explaining variation in health status across space and time: implications for racial and ethnic disparities in self-rated health. AB - We use the Metropolitan Community Information Center-Metro Survey-a serial cross section of adults residing in the City of Chicago, USA, conducted from 1991 through 1999-in combination with 1990 census data to simultaneously examine the extent to which self-rated health varies across Chicago neighborhoods and across time. Three-level hierarchical logit models are employed to decompose individual, spatial, and temporal variance in self-rated health. Results indicate that variation in self-rated health across neighborhoods is explained, in part, by variation in the level of neighborhood affluence. Neighborhood level poverty, however, is not a significant predictor of self-rated health. Community level affluence, moreover, accounts for a substantial proportion of the residual health deficit experienced by African-Americans when compared with Whites (after controlling for individual level SES). The effects of affluence hold when controlling for spatial autocorrelation and when considered in primarily African American neighborhoods. Findings also indicate that individuals living in the City of Chicago became significantly healthier over the decade of the 1990s, and that this improvement in health is explained largely by the increasing education and income levels of Chicago residents. PMID- 12899907 TI - Do age and social relations moderate the relationship between self-rated health and mortality among adult Danes? AB - It is well established that self-rated health (SRH) predicts mortality even after controlling for a wide range of factors. We explored the extent to which age and social relations (structural and functional) influenced the relationship between SRH and mortality (after 13 years follow-up) in a representative sample of adult Danes (N=6693). After controlling for socioeconomic status, illness, and lifestyle variables, we found that age moderated the SRH-mortality relationship such that it was present for respondents under 55 but absent for respondents over 56. In addition, weaker structural (but not functional) social relations increased mortality directly but neither structural nor functional social relations moderated the SRH-mortality relationship. We discuss the theoretical and practical implications of these findings. PMID- 12899908 TI - Predictors of psychological distress in Lebanese hostages of war. AB - A cross-sectional study addressed the experience of Lebanese hostages of war in Lebanon. It specifically assessed the prevalence of general distress and its relationship to captivity-related factors and selected psychosocial variables. Trained field researchers using standard measurements interviewed 118 Lebanese hostages released from Khiam prison, an Israeli detention center in Lebanon. Questionnaires administered included the GHQ-12 and the Harvard Trauma Questionnaire. Individuals were detained for 3.4 years on average, and 86% were tortured. Psychological distress was present in 42.1% of the sample compared to 27.8% among the control group. In the multivariate analysis, the significant predictors for distress were: years of education and increase in religiosity after release. In conclusion, even after 2 years of release, more than one-third of the Lebanese hostages released from Khiam prison were found to have psychological distress. Caregivers need to pay special attention to the mental health of hostages of war. The paper discusses the meaning and implications of the factors predicting resilience and vulnerability in this particular population. PMID- 12899909 TI - Preliminary development of the World Health Organsiation's Quality of Life HIV instrument (WHOQOL-HIV): analysis of the pilot version. AB - The assessment of quality of life (QoL) is central to understanding how people's lives are affected by HIV infection. A reliable and valid measurement tool developed for cross-cultural use will be important in evaluating the global impact of the disease. This paper reports on the development and preliminary assessment of the WHOQOL-HIV pilot instrument that is designed for use with the WHOQOL-100 for persons living with HIV and AIDS (PLWHA). In this study, 900 people with a mean age of 32 from six culturally diverse sites completed the WHOQOL-100 along with 115 HIV specific items. Respondents were HIV asymptomatic (23%), HIV symptomatic (23%), had AIDS (20%) or were well (34%). Analyses to select the best items from the piloted instrument resulted in the inclusion of 33 items covering 12 new facets for a field trial version of the WHOQOL-HIV instrument; e.g. symptoms of HIV, body image, social inclusion, death and dying, and forgiveness. The results indicate excellent internal consistency for the scale (alpha=0.98) and its domains (alpha=0.87-0.94). For PLWHA, pain and discomfort, positive feelings, dependence on medication, sexual activity, financial resources and spiritual connection were particularly poor, indicating that the severest impact of HIV extends beyond physical well-being to the psycho social-spiritual and environmental areas of QoL. Comparisons using ANOVA showed that persons who are at later stages of HIV infection, or are currently ill report poorer QoL than those that were well (p<0.01). Women report poorer QoL than men for almost every facet (p<0.01) and older persons (>30) reported lower negative feelings, and better social inclusion, spiritual connection, forgiveness and spiritual experience than younger persons. Finally, those with no education, or only primary education showed some of the poorest means. It is concluded that these new items and facets add value for measurement of QoL in PLWHA. PMID- 12899910 TI - Predicting young children's quality of life. AB - This paper represents an investigation into the determinants of young children's quality of life (QOL) in Thailand. The empirical work is based upon a sample of 498 children (aged 5-8); 220 were urban children and 278 children of construction workers in Bangkok. Their QOL was assessed using a new self-report QOL measure for children. Multiple regression analyses indicated that the father's income and education, type of school, mode of transportation to school, and the amount of time that the child spent on extra study courses were significant explanatory variables. It was found that these factors had different influences on the QOL of urban children and those of construction workers. Extra sport-related activities and extra work (other than housework) improved the QOL of urban children, while the QOL of construction workers' children was directly linked to father's education and income. This result is consistent with income having a diminishing marginal effect on the QOL of children. There is also evidence that amongst construction workers' children, boys have a lower QOL than girls. The different causal explanations for the QOL of urban and construction workers' children suggests that it is context specific, and what impacts one group of children's QOL within a particular context may not impact another group in a different situation. This has important policy implications. Throughout the study we could find no significant impact of health on QOL-neither chronic, acute nor severe illness has any significant impact on QOL. This is consistent with the hypothesis that QOL is influenced by expectations (Social Science and Medicine 41 (10) (1995) 1403). Findings of the effects of social and environmental factors on children's QOL are new in this field and should be further investigated. PMID- 12899911 TI - A systematic and critical review of the process of translation and adaptation of generic health-related quality of life measures in Africa, Asia, Eastern Europe, the Middle East, South America. AB - In recent years there has been a worldwide increase in demand for subjective measures of health-related quality of life (HRQL). Researchers have the choice of whether to develop a new measure or whether to adapt an existing measure in another language. This review evaluates the processes used in translating and adapting nine generic HRQL instruments (15D, Dartmouth COOP/WONCA Charts, EuroQol, HUI, NHP, SIP, SF-36, QWB, WHOQOL) for use in Africa, Asia, Eastern Europe, the Middle East, and South America. The review adopts a universalist model of equivalence, outlined by Herdman, Fox-Rushby, and Badia (Qual. Life Res. 7 (1998) 323), to judge the 58 papers reviewed. Research spans 23 countries and is dominated by research in the East Asia and Pacific region and the SF-36. Results are reported for conceptual, item, semantic, operational, measurement and functional equivalence. It is argued that currently there is a misguided pre occupation with scales rather than the concepts being scaled and too much reliance on unsubstantiated claims of conceptual equivalence. However, researchers using the WHOQOL approach are more likely to establish reliable conclusions concerning the equivalence of their instrument across countries. It is a key conclusion of this review that research practice and translation guidelines still need to change to facilitate more effective and less biased assessments of equivalence of HRQL measures across countries. PMID- 12899912 TI - Daily geographies of caregivers: mobility, routine, scale. AB - The bulk of care provided to elderly people living in the community and needing assistance is provided informally by family and friends. This paper investigates themes from an interpretation of interviews with informal caregivers about their experiences of caring for a frail, ill, or disabled elderly person at home. These themes include mobility, routine, and inter-relationships of scale. The caregivers' narratives illustrate the interconnected nature of physical, material and social, emotional aspects of care, and the profound spatial and social impact of providing informal care to a family member on their everyday lives. These are often at odds with political and social constructions of what it means to care at home. Concepts of space, place, and time are shown to be a helpful framework through which to understand issues and experiences of caring. The social and the physical aspects of the many interconnected scales and places which caregivers negotiate on an everyday basis both shape and are shaped by caregiving. PMID- 12899913 TI - Women doctors in Norway: the challenging balance between career and family life. AB - In most Western countries, women doctors are still underrepresented in the higher positions in the medical hierarchy and in the most prestigious specialities. A crucial question is whether family responsibilities affect female and male career differently. The article examines how Norwegian physicians balance their work and family responsibilities and demonstrates differences in the way doctors combine work and family obligations, between women and compared with men. Among women doctors, the probability of becoming a specialist decreased with an increasing number of children. Moreover, postponing the birth of the first child increased the probability of completing hospital specialities. Although more women than men work part-time, this was the case only for a small proportion of women doctors. Transition from full-time to part-time work is primarily an accommodating strategy to family responsibilities, however strongly influenced by variations in the opportunity structure of different specialities. The findings further demonstrate that being married to another doctor had a positive impact on the career, especially for women doctors. PMID- 12899914 TI - Opioid receptor interactions: local and nonlocal, symmetric and asymmetric, physical and functional. AB - The pharmacological effects of opioid drugs and endogenous opioid peptides are mediated by several kinds of receptors, the major ones being mu, delta and kappa. Though classically it has been thought that a particular effect mediated by a drug or other ligand results from its interaction with a single type of receptor, accumulating evidence demonstrates that interactions between receptors play a major role in opioid actions. These interactions may be either local, involving receptors within the same tissue, or nonlocal, between receptors located in different tissues. Nonlocal interactions always involve intercellular mechanisms, whereas local interactions may involve either intercellular or intracellular interactions, the latter including physical association of receptors. Both local and nonlocal interactions, moreover, may be either symmetric, with ligand interaction at one receptor affecting interaction at the other, or asymmetric; and either potentiating or inhibitory. In this article we discuss major examples of these kinds of interactions, and their role in the acute and chronic effects of opioids. Knowledge of these interactions may have important implications for the design of opioids with more specific actions, and for eliminating the addictive liability of these drugs. PMID- 12899915 TI - Induction of apoptosis by takrisodokyeum through generation of hydrogen peroxide and activation of caspase-3 in HL-60 cells. AB - Takrisodokyeum (TRSDY), a Chinese herbal medicine, has been known to exert anti tumoral activity in Korea. However, its molecular mechanism of action is not understood. In this study, we found that TRSDY induced apoptosis in HL-60 cells as evidenced by both a characteristic ladder pattern of discontinuous DNA fragments and an increase of annexin V+/PI- stained cell population. Our data demonstrated that TRSDY-induced apoptotic cell death was accompanied by activation of caspase-3 and cleavages of its substrates, poly(ADP-ribose) polymerase (PARP) and RhoGDP dissociation inhibitor (RhoGDI-2; also called D4 GDI) in a time- and concentration-dependent manner. Caspase-3 inhibitor, but not caspase-1 inhibitor, prevented TRSDY-induced apoptosis. Furthermore, treatment with TRSDY increased the production of intracellular hydrogen peroxide and pretreatment of cells with anti-oxidants conferred complete protection against hydrogen peroxide generation and subsequent caspase-3 activation. Taken together, these results suggest that TRSDY induces hydrogen peroxide generation, which, in turn, causes activation of caspase-3, degradation of PARP and D4-GDI, and eventually leads to apoptotic cell death. PMID- 12899916 TI - Altered expression of NCAM in hippocampus and cortex may underlie memory and learning deficits in rats with streptozotocin-induced diabetes mellitus. AB - Neurological and structural changes are paralleled by cognitive deficits in diabetes mellitus. The present study was designed to evaluate the expression of neural cell adhesion molecules (NCAM) in the hippocampus, cortex and cerebellum and to examine cognitive functions in diabetic rats. Diabetes was induced in male albino rats via intraperitoneal streptozotocin injection. Learning and memory behaviors were investigated using a passive avoidance test and a spatial version of the Morris water maze test. NCAM expression was detected in the hippocampus, cortex and cerebellum by an immunoblotting method. The diabetic rats developed significant impairment in learning and memory behaviours as indicated by deficits in passive avoidance and water maze tests as compared to control rats. Expression of NCAM 180 and 120 kDa were found to be higher in hippocampus and cortex of diabetic rat brains compared to those of control, whereas expression of NCAM 140 kDa decreased in these brain regions. Our findings suggest that streptozotocin induced diabetes impairs cognitive functions and causes an imbalance in expression of NCAM in those brain regions involved in learning and memory. Altered expression of NCAM in hippocampus may be an important cause of learning and memory deficits that occur in diabetes mellitus. PMID- 12899917 TI - Antinociception induced by beta-lactotensin, a neurotensin agonist peptide derived from beta-lactoglobulin, is mediated by NT2 and D1 receptors. AB - In this study, we examined the antinociceptive effect of beta-lactotensin, a neurotensin agonist that has been isolated from the chymotrypsin digest of beta lactoglobulin as an ileum-contracting peptide. Beta-lactotensin showed naloxone insensitive antinociceptive activity by the tail-pinch test after i.c.v. (200 nmol/mouse) or s.c. (300 mg/kg) administration in ddY mice. Tolerance was not developed to antinociception induced by beta-lactotensin after repeated s.c. administration for 5 days. The antinociceptive activity of beta-lactotensin was blocked by treatment with the neurotensin NT2 receptor antisense ODN, while treatment with the NT1 receptor antisense ODN had no effect. The antinociceptive activity was also blocked by a dopamine D1 receptor antagonist, SCH23390 (1 microg/mouse, i.c.v.), while a D2 receptor antagonist, raclopride (0.5 microg/mouse, i.c.v.), did not block the activity. These results indicate that the antinociceptive activity of beta-lactotensin is mediated by NT2 and D1 receptors. PMID- 12899918 TI - Antihyperglycemic effect of aqueous and ethanolic extracts of Gongronema latifolium leaves on glucose and glycogen metabolism in livers of normal and streptozotocin-induced diabetic rats. AB - The present study was designed to investigate the antihyperglycemic effects of aqueous and ethanolic extracts from Gongronema latifolium leaves on glucose and glycogen metabolism in livers of non-diabetic and streptozotocin-induced diabetic rats. To investigate the effects of aqueous or ethanolic leaf extracts of G. latifolium, non-diabetic and STZ diabetic rats were treated twice daily (100 mg/Kg) for two weeks. Diabetic rats showed a significant decrease in the activities of hepatic hexokinase (HK), phosphofructokinase (PFK) and glucose-6 phosphate dehydrogenase (G6PDH) and an increase in glucokinase (GK) activity. The levels of hepatic glycogen and glucose were also increased in diabetic rats. However, there were no significant differences in the activities of glucose-6 phosphatase (G6Pase) in treated and untreated diabetic rats. The ethanolic extract significantly increased the activities of HK (p<0.01), PFK (p<0.001) and G6PDH (p<0.01) in diabetic rats, decreased the activity of GK (p<0.05) and the levels of hepatic glycogen (p<0.01) and both hepatic (p<0.001) and blood glucose (40%). The aqueous extract of G. latifolium was only able to significantly increase the activities of HK and decrease the activities of GK but did not produce any significant change in the hepatic glycogen and both hepatic and blood glucose content of diabetic rats. Our data show that the ethanolic extract from G. latifolium leaves has antihyperglycemic potency, which is thought to be mediated through the activation of HK, PFK, G6PDH and inhibition of GK in the liver. The ethanolic extract is under further investigation to determine the chemical structure of the active compound(s) and its/their mechanism of action. PMID- 12899919 TI - Tachykinins mediate non-adrenergic, non-cholinergic excitatory neurotransmission to the hamster ileum via NK1 and NK2 receptors. AB - The present study was designed to investigate Substance P (SP) and a related tachykinin, Neurokinin A (NKA), contributions to the excitatory neurotransmission to the circular smooth muscle of the hamster ileum. In the presence of atropine (0.5 microM), guanethidine (3 microM) and NG-nitro-L-arginine methyl ester (L NAME) (200 microM), electrical field stimulation (EFS) evoked a non-adrenergic, non-cholinergic (NANC) excitatory junction potential (EJP) and contraction of circular smooth muscle. Applications of SP and NKA produced depolarizing and contractile responses in a concentration-dependent fashion. The EJP and contraction were almost abolished by the non-specific tachykininergic antagonist, spantide (3 microM). Application of SP antagonist, L-732,138, (1 microM) markedly inhibited EJP (82.5%) and contraction (68.9%) and completely blocked excitatory responses produced by exogenous application of SP. While application of NKA antagonist, SR48968 (1 microM) completely blocked the depolarising and contractile responses to NKA, it only slightly inhibited those to EFS (17.2% and 31.4% respectively). These results provide evidence that, in the circular muscle of hamster ileum, endogenous tachykinins are the main NANC excitatory neurotransmitters and their action is mediated by both NK1 and NK2 receptors. PMID- 12899920 TI - Abnormal brain activation to visual stimulation in cocaine abusers. AB - Chronic cocaine abuse has been associated with cerebrovascular pathology. This is likely to reflect its vasoactive effects; cocaine produces vasoconstriction and reduces cerebral blood flow. We propose that cerebrovascular pathology in chronic cocaine abusers would result in abnormal BOLD [blood oxygenation level dependent] responses to activation stimuli. Here, we used fMRI to compared the BOLD response to photic visual stimulation in neurologically intact active cocaine abusers to that in non-drug-using healthy controls. Cocaine abusers showed a significantly enhanced positive BOLD response to photic stimulation when compared to control subjects. The enhanced activation in the cocaine abusers could result from low resting cerebral blood flow secondary to increased vasoconstriction and/or from low oxidative metabolism during activation. Alternatively, the larger signal intensity in the cocaine abusers could result from inefficient neuronal processing as has been shown to occur in other conditions of cerebral pathology. These findings provide evidence of cerebral dysfunction with chronic cocaine abuse, which could reflect cerebral blood flow or neuronal changes. Further studies are required to determine if the cerebrovascular changes we observed in the cocaine abusers recover with detoxification and to assess their functional consequences. PMID- 12899921 TI - The multiple promoter methylation profile of PR gene and ERalpha gene in tumor cell lines. AB - The changes of methylation status of various gene promoters are a common feature of malignant cells and these changes can occur early in the progression process. Therefore, abnormal methylation can be used as cancer marker. Such studies will first require the development of a panel of methylated markers that are methylated in cancer tissues but unmethylated in normal tissues or methylated status is different between cancer tissues and normal tissues. By using methylation-specific PCR (MSP) assay method, we observed alterations in DNA methylation at the double promoter regions of the progesterone receptor (PR) gene and estrogen receptor (ERalpha) gene in various tumor cell lines. Compared with normal white blood cell, the methylation status of PRA promoter in various cancer cell lines changed from unmethylation pattern to methylation pattern. That of PRB promoter changed from both unmethylated and methylated alleles to only methylated allele. The methylation status of ERalpha-A and ERalpha-B promoter in various cancer cell lines are cell -specific. This study indicates that PR promoter methylation may be a molecular marker in various cancer detections. And the methylation status of ERalpha-A and ERalpha-B is cell-specific. PMID- 12899922 TI - Polydeoxyribonucleotide (PDRN) promotes human osteoblast proliferation: a new proposal for bone tissue repair. AB - Several researchers have recently shed new light upon the importance of extracellular nucleotides and nucleosides to stimulate cells growth. PDRN, a mixture of deoxyribonucleotides polymers of different lengths, has recently demonstrated to stimulate "in vitro" fibroblast proliferation and collagen production, probably stimulating the purinergic receptor system. In this work we evaluated the effects of PDRN on human cultured osteoblasts, focusing our attention on cell proliferation and alkaline phosphatase activity. PDRN at a concentration of 100 microg/ml induce an increase in osteoblasts growth after 6 days as compared to control (+21%). The addition of DMPX 50 microM and suramine (P2 inhibitor) 10 microM give different results: suramine has no significant effect, while DPMX reduce, even if partially, the PDRN induced cell growth. The alkaline phosphatase activity shows a gradual enhancement starting from day 0 to day 10, even if PDRN treated cells, examined at day 6, present a sensibly lower phosphatase activity when compared to controls. Our data demonstrate that PDRN acts as an osteoblast growth stimulator. Its action is partially due to a stimulation of the purinergic system mediated by A2 purinoreceptors, however we can not exclude the involvement of other mechanism like salvage pathway. PMID- 12899923 TI - Angiogenesis and nerve regeneration in a model of human skin equivalent transplant. AB - The angiogenesis and reinnervation were studied in a porcine model of human skin equivalent (SE) graft and the relationship between the two processes was investigated. Confocal laser scanning microscopy was used to monitor, during the healing process, the pattern of vascularization and reinnervation at different time points. The SE was obtained by co-culturing fibroblasts and keratinocytes on a collagen-glycosaminoglycan-chitosan biopolymer and grafted on dorsal wounds generated by full-thickness resection in 25/30 Kg Large white pigs. Frozen sections were obtained from biopsies performed in autograft and xenograft, then were immunolabeled by using the endothelial marker lectin Lactifolia and with the neuronal marker gene product PGP9.5. Cajal staining was also used to visualize the nerve fibers. The results show that the vascularization precedes the innervation process. These data are consistent with the view that the development of nervous tissue is driven by nutritional and trophic factors provided by the vascular system. The arborization of the two systems observed during the third week from the graft might play a key role in maintaining the healing process and the graft survival. PMID- 12899924 TI - The antinociceptive effect of iontophoretic direct application of diclofenac to arthritic knee-joints of rats. AB - This study compared the antinociceptive effect produced by cathodic iontophoresis of sodium diclofenac close to an arthritic knee-joint in rats with that of systemic application. Arthritic nocifensive incapacitation was induced by LPS (1 microg) injection into a knee-joint previously (72 h) primed with carrageenan (300 microg). Diclofenac (0.1, 0.25 and 0.5 mg/kg) given intraperitoneally 1 h after LPS injection caused dose-dependent inhibition of incapacitation. Diclofenac iontophoresis was performed by varying either the current density (0.1, 0.2, and 0.3 mA/cm2) or the duration of application (4, 10, 20 and 30 min) of a polyvinylpirrolydone-hydroxymethylcellulose gel containing 1% sodium diclofenac. A clear, current density-dependent effect was observed for 0.1, 0.2 and 0.3 mA/cm2 (10 min period), which was similar to the effect observed for the intraperitoneal application of 0.1, 0.25 and 0.5 mg/kg doses. Combining different application periods with different current densities, in a manner that resulted in the same total current (1.6 mA*min) application, did not produce similar therapeutic effects, but the antinociceptive effect was directly proportional to the current density. The ipsilateral iontophoresis (0.25 mA/cm2 x 10 min or 0.5 mA/cm2 x 4 min) of diclofenac produced an effect significantly greater than the same contralateral application (p<0.05). In conclusion, our results suggest that the therapeutic effect depends on the current density but not on the application time, and also that the iontophoretic, direct application to the inflamed knee joint enhances the therapeutic effect probably as a result of the direct delivery of the drug. PMID- 12899925 TI - Utilization of in vivo ultrafiltration in biomedical research and clinical applications. AB - Ultrafiltration (UF) is a filtrate selection method with a wide range of biomedical and clinical applications, including detoxification of blood in hemodialysis and peritoneal dialysis. New is, however, the use of UF as a convenient in vivo sampling method that, for example, has been used in diabetics. Ultrafiltration avoids complicated and time-consuming recovery calculations that are necessary when using in vivo microdialysis, as recoveries of low molecular weight molecules are near 100%. The subcutaneously or intravenously placed UF probes have been studied for off-line sample analysis and for continuous on-line monitoring, in a wide variety of species, including dogs, rats, pigs and humans. This review discusses the potential of in vivo UF as a continuous tissue sampling technique in clinical research areas, and in several major biomedical applications including glucose and lactate monitoring and drug kinetic studies. PMID- 12899926 TI - cGMP stimulates endoplasmic reticulum Ca(2+)-ATPase in vascular endothelial cells. AB - Calcium is a crucial regulator of many physiological processes such as cell growth, division, differentiation, cell death and apoptosis. In this study, we examined the effect of cGMP on agonist-induced [Ca(2+)](i) transient in isolated rat aortic endothelial cells. 100 microM ATP was applied to the cells bathed in a Ca(2+)-free physiological solution to induce a [Ca(2+)](i) transient that was caused by Ca(2+) release from intracellular stores. cGMP, which was applied after [Ca(2+)](i) reached its peak level, accelerated the falling phase of [Ca(2+)](i) transient. Pre-treatment of the cells with CPA abolished the accelerating effect of cGMP on the falling phase of [Ca(2+)](i) transient. The effect of cGMP was reversed by KT5823, a highly specific inhibitor of protein kinase G. Taken together, these data suggest that cGMP may reduce [Ca(2+)](i) level by promoting Ca(2+) uptake through sarcoplasmic/endoplasmic reticulum ATPase and that the effect of cGMP may be mediated by protein kinase G. PMID- 12899927 TI - Major royal jelly protein 3 modulates immune responses in vitro and in vivo. AB - We have recently shown that royal jelly has potent antiallergic properties in a mouse model of immediate hypersensitivity. However, it is still unclear which components of royal jelly exhibit antiallergic activity. In this study, we have screened for antiallergic factors in royal jelly based on inhibition of IL-4 production by anti-CD3 stimulated spleen cells derived from OVA/alum-immunized mice. Using a series of column chromatographies, we purified a 70 kDa glycoprotein, major royal jelly protein 3 (MRJP3), that suppresses IL-4 production. In in vitro experiments, MRJP3 suppressed the production of not only IL-4 but also that of IL-2 and IFN-gamma by T cells concomitant with inhibition of proliferation. The MRJP3-mediated suppression of IL-4 production was also evident when lymph node cells from OVA/alum-immunized mice were stimulated with OVA plus antigen presenting cells. We next examined the purified suppressive factor on OVA/alum-induced allergic responses in mice. Interestingly, in spite of the antigenicity of MRJP3 itself as an extraneous foreign protein, intraperitoneal administration of MRJP3 inhibited serum anti-OVA IgE and IgG1 levels in immunized mice. In addition, heat-treated soluble MRJP3 treatment reduced its antigenicity while maintaining its inhibitory effects on antibody responses to OVA. These results indicate that MRJP3 can exhibit potent immunoregulatory effects in vitro and in vivo. Furthermore, considering the intriguing immunomodulatory effects of MRJP3, it may be of clinical significance to design MRJP3-derived antiallergic peptides by identifying the associated polypeptide regions. PMID- 12899928 TI - Reactive oxygen species mediate doxorubicin induced p53-independent apoptosis. AB - Doxorubicin (DOX) is a common anticancer drug. The mechanisms of DOX induced apoptosis and the involvement of reactive oxygen species (ROS) in apoptotic signaling were investigated in p53-null human osteosarcoma Saos-2 cells. Accumulation of pre-G1 phase cells and induction of DNA laddering, which are the hallmarks of apoptosis, were detected in cells at 48 h upon DOX treatment. Furthermore, DOX increased the intracellular hydrogen peroxide and superoxide levels, followed by mitochondrial membrane depolarization, cytochrome c release, caspase-3 activation, prior to DNA laddering in Saos-2 cells. In addition, DOX treatment also upregulated Bax and downregulated Bcl-2 levels in the cells. The role of ROS in DOX induced cell death was confirmed by the suppression effect of catalase on DOX induced ROS formation, mitochondrial cytochrome c release, procaspase-3 cleavage, and apoptosis in Saos-2 cells. The catalase treatment however only suppressed DOX induced Bax upregulation but had no effect on Bcl-2 downregulation. Results from the present study suggested that ROS might act as the signal molecules for DOX induced cell death and the process is still functional even in the absence of p53. PMID- 12899929 TI - The effect of natural clinoptilolite on the serotonergic receptors in the brain of mice with mammary carcinoma. AB - The ex vivo effect of tribomechanically micronized zeolite (MZ) on the binding of 3H-8-OH-DPAT to 5-HT(1A) and 3H-5-HT to 5-HT(1B) receptors was investigated in the brain of nontumorous (control) and mammary carcinoma bearing female mice. During 14 and 28 days mice were fed with standard food, standard food supplemented with 25% of MZ, or standard food supplemented with 25% of non tribomechanically micronized zeolite (non-MZ). A reduced binding of 3H-8-OH-DPAT to 5-HT(1A) receptors in mammary carcinoma bearing mice was found when compared to control mice fed with standard food for 28 days, suggesting a time dependent alteration of 5-HT(1A) receptors in mammary carcinoma. The addition of MZ for 28 days in these mice abolished the decrease in 5-HT(1A) receptors binding, indicating a possible beneficial effect of MZ, at least on 5-HT(1A) receptors in mammary carcinoma bearing mice. The preliminary data show that MZ administered as a food supplement (25%) for 14 days induced a transient decrease in the binding of 3H-5-HT to brain 5-HT(1B) receptors only in control, but not in tumor-bearing mice, that disappeared after 28 days of MZ-supplemented food administration. The mechanism of the indirect action of MZ on the brain serotonergic receptors might be achieved by the alterations in the electrolytes balance, and/or by the regulation of the immune system. PMID- 12899930 TI - ATP and ADP hydrolysis in brain membranes of zebrafish (Danio rerio). AB - Nucleotides, e.g. ATP and ADP, are important signaling molecules, which elicit several biological responses. The degradation of nucleotides is catalyzed by a family of enzymes called NTPDases (nucleoside triphosphate diphosphohydrolases). The present study reports the enzymatic properties of a NTPDase (CD39, apyrase, ATP diphosphohydrolase) in brain membranes of zebrafish (Danio rerio). This enzyme was cation-dependent, with a maximal rate for ATP and ADP hydrolysis in a pH range of 7.5-8.0 in the presence of Ca(2+) (5 mM). The enzyme displayed a maximal activity for ATP and ADP hydrolysis at 37 degrees C. It was able to hydrolyze purine and pyrimidine nucleosides 5'-di and triphosphates, being insensitive to classical ATPase inhibitors, such as ouabain (1 mM), N ethylmaleimide (0.1 mM), orthovanadate (0.1 mM) and sodium azide (0.1 mM). A significant inhibition of ATP and ADP hydrolysis (68% and 34%, respectively) was observed in the presence of 20 mM sodium azide, used as a possible inhibitor of ATP diphosphohydrolase. Levamisole (1 mM) and tetramisole (1 mM), specific inhibitors of alkaline phosphatase and P1, P(5)-di (adenosine 5'-) pentaphosphate, an inhibitor of adenylate kinase did not alter the enzyme activity. The presence of a NTPDase in brain membranes of zebrafish may be important for the modulation of nucleotide and nucleoside levels, controlling their actions on specific purinoceptors in central nervous system of this specie. PMID- 12899931 TI - Dietary alpha-linolenic acid suppresses the formation of lysophosphatidic acid, a lipid mediator, in rat platelets compared with linoleic acid. AB - Rats fed a high linoleic acid (LA, 18:2n-6) diet or a high alpha-linolenic acid (ALA, 18:3n-3) diet for 4 months after weaning. Platelets from the high-LA group contained more arachidonic acid (AA, 20:4n-6) and less eicosapentaenoic acid (EPA, 20:5n-3) and docosahexaenoic acid (DHA, 22:6n-3) compared with those from the high-ALA group. Incorporation of [32P]orthophosphate into platelet phospholipids was increased by thrombin-treatment, and was greater by ca. 30% in the high-LA group than in the high-ALA group both in the presence and absence of thrombin. The formation of [32P]lysophosphatidic acid (LPA), a lipid messenger, in [32P]orthophosphate-labeled platelets was increased 6.6-fold in the high-LA group and 4.1-fold in the high-ALA-group by thrombin-treatment. The formation of [32P] LPA in activated platelets was reduced by 35% in the high-ALA group. PMID- 12899932 TI - 2'-benzyloxychalcone derivatives stimulate glucose uptake in 3T3-L1 adipocytes. AB - By a cell-based glucose uptake screening assay, a chalcone derivative, 3-nitro-2' benzyloxychalcone (compound 1) was identified. Compound 1 stimulated glucose uptake and potentiated insulin-stimulated glucose uptake in a concentration dependent manner in 3T3-L1 adipocytes. When cells were treated with various concentrations of insulin in the presence of compound 1, marked enhancement of insulin-stimulated glucose uptake was observed at each concentration, suggesting that the compound might function as an insulin sensitizer. Preliminary study on the structure-activity relationships revealed that two aromatic benzene rings tolerated several substituents, but substitution by acidic or highly polar groups abolished the activity. Among several chalcone derivatives, 4-chloro-2' benzyloxychalcone (compound 8) showed the highest level of activity. Compound 8 stimulated glucose uptake was almost completely inhibited by wortmannin, a specific inhibitor of phosphatidylinositol 3-kinase (PI3K). These results suggest that the action of chalcone derivatives is mediated via a pathway involving PI3K. PMID- 12899933 TI - Effects of IgM from rheumatic fever patients on intracellular calcium levels of neonatal rat cardiac myocytes. AB - Rheumatic fever (RF), a potential sequela of Streptococcus pyogenes pharyngitis, sometimes results in myocarditis and heart failure. Antibodies have been implicated in the pathogenesis of RF and anti-cardiac myosin antibody levels are elevated in RF patients. Since myocarditis is associated with altered cardiomyocyte calcium transients it was of interest to determine the direct effects of RF patient antibodies on calcium transients in cultured myocytes. RF patient polyclonal IgM treatment caused increased calcium retention by neonatal rat heart cells in vitro as determined with isotopically labeled calcium. Therefore, to further characterize this finding, calcium transients were evaluated by real time fluorescence spectroscopy and deconvolution imaging. RF patient polyclonal IgM produced increased calcium retention during the relaxation stage of the contraction cycle leading to a slowing of contraction rate, disorganized calcium transients, and eventual tetany. In contrast, calcium transient studies of cardiomyocytes following treatment with monoclonal anti myosin antibodies revealed declining intracellular calcium levels, accompanied by disorganized transients and tetany. Treatment with both antibodies led to myocyte dysfunction and these novel findings suggest a role for antibodies in the pathogenesis of the myocarditis associated with rheumatic carditis. PMID- 12899934 TI - Effects of sensorial denervation induced by capsaicin injection at birth or on day three of life, on puberty, induced ovulation and pregnancy. AB - Evidence that ovarian innervation plays a role in the regulation of ovarian functions has been widely reported. The present study analyzed the effects of treating neonatal rats with capsaicin on: the onset of puberty, serum hormone levels, timing of first vaginal estrus, ovulation rates, ovarian and uterine weights, follicular development, and noradrenaline concentration in the ovaries. The study was based on two experimental models: untouched rats and hemispayed animals. The results indicate that newborns treated with capsaicin did not show changes in the onset of puberty, though the number of ova shed by these animals was lower than in control rats. The number of follicles counted in the ovaries of newborn, or 3-day old-capsaicin-treated animals, was significantly lower and showed an increase in the number of atretic follicles compared to vehicle treated rats. All capsaicin treated animals copulated, but only 6/23 delivered pups. The number of ova shed by capsaicin-treated animals after eCG or eCG-hCG treatment was significantly lower than in the control group. Present results could indicate that some specific neural information registered in the ovary is sent to the central nervous system, and that such information plays a role in the mechanism regulating ovarian function. PMID- 12899935 TI - In vivo and in vitro stimulatory effects of Cordyceps sinensis on testosterone production in mouse Leydig cells. AB - The in vivo and in vitro effects of Cordyceps sinensis (CS) and its extracted fractions on the secretion of testosterone in mice were studied. CS, F2 (water soluble protein), and F3 (poorly water soluble polysaccharide and protein) significantly stimulated in vitro testosterone production in purified mouse Leydig cells. However, F1 (water soluble polysaccharide) had no effect (p>0.05). F2 and F3 stimulated in vitro testosterone production in dose- and time-dependent relationships with maximal responses at 3 mg/ml for 3 h (p<0.05). An in vivo study illustrated that testosterone levels in plasma were significantly increased by CS, F2, and F3, respectively (p<0.05). Because CS, F2, and F3 stimulated both in vitro and in vivo testosterone secretions in mice, it is possible that CS might contribute to an alternative medicine for the treatment of some reproductive problems caused by insufficient testosterone levels in human males. PMID- 12899936 TI - Reactive oxygen species in pulmonary inflammation by ambient particulates. AB - Exposure to ambient air pollution particles (PM) has been associated with increased cardiopulmonary morbidity and mortality, particularly in individuals with pre-existing disease. Exacerbation of pulmonary inflammation in susceptible people (e.g., asthmatics, COPD patients) appears to be a central mechanism by which PM exert their toxicity. Health effects are seen most consistently with PM with aerodynamic diameter < 2.5 micrometers (PM(2.5)), although 10 micrometers < PM < 2.5 micrometers can also be toxic. Through its metal, semi-quinone, lipopolysaccaride, hydrocarbon, and ultrafine constituents, PM may exert oxidative stress on cells in the lung by presenting or by stimulating the cells to produce reactive oxygen (ROS). In vivo, PM increase cytokine and chemokine release, lung injury, and neutrophil influx. In vitro analysis of PM effects on the critical cellular targets, alveolar macrophages, epithelial cells, and neutrophils, demonstrates PM- and oxidant-dependent responses consistent with in vivo data. These effects have been observed with PM samples collected over years as well as concentrated PM(2.5) (CAPs) collected in real time. Oxidative stress mediated by ROS is an important mechanism of PM-induced lung inflammation. PMID- 12899937 TI - Pathways of cell signaling in hyperoxia. AB - Administration of high concentrations of oxygen (hyperoxia) is a mainstay of supportive treatment for patients suffering from severe respiratory failure. However, hyperoxia, by generating excess systemic reactive oxygen species (ROS), can exacerbate organ failure by causing cellular injury. Therefore, a better understanding of the signal transduction pathways in hyperoxia may provide the basis for effective therapeutic interventions. The major biological effects of hyperoxia include cell death, induction of stress responses, inflammation, and modulation of cell growth. Major signaling pathways that appear to be involved include the mitogen-activated protein kinases (MAPKs), AP-1, and NF-kappa B, which converge, ultimately, to the expression of a range of stress response genes, cytokines, and growth factors. PMID- 12899938 TI - Pharmacological rescue of the 14CoS/14CoS mouse: hepatocyte apoptosis is likely caused by endogenous oxidative stress. AB - Whereas ch/ch wild-type mice and ch/14CoS heterozygotes are viable, 14CoS/14CoS mice homozygous for a 3800 kb deletion on chromosome 7 die during the first day postpartum. Death is caused by disruption of the fumarylacetoacetate hydrolase (Fah) gene; absence of FAH, final enzyme in the tyrosine catabolism pathway, leads to accumulation of reactive electrophilic intermediates. In this study, we kept 14CoS/14CoS mice alive for 60 d with oral 2-(2-nitro-4-trifluoromethyl benzyol)-1,3-cyclohexanedione (NTBC), an inhibitor of p-hydroxyphenylpyruvate dioxygenase, second enzyme in the tyrosine catabolic pathway. The 70% of NTBC treated 14CoS/14CoS mice that survived 60 d showed poor growth and developed corneal opacities, compared with ch/14CoS littermates; NTBC-rescued Fah(-/-) knockout mice did not show growth retardation or ocular toxicity. NTBC-rescued 14CoS/14CoS mice also exhibited a striking oxidative stress response in liver and kidney, as measured by lower GSH levels and mRNA induction of four genes: glutamate cysteine ligase catalytic (Gclc) and modifier (Gclm) subunits, NAD(P)H:quinone oxidoreductase (Nqo1), and heme oxygenase-1 (Hmox1). Withdrawal of NTBC for 24-48 h from rescued adult 14CoS/14CoS mice resulted in severe apoptosis of the liver, detected histologically and by cytochrome c release from the mitochondria, increased caspase 3-like activity, and further decreases in GSH content. In kidney, proximal tubular epithelial cells were abnormal. Human hereditary tyrosinemia type I (HT1), caused by mutations in the FAH gene, is an autosomal recessive disorder in which the patient usually dies of liver fibrosis and cirrhosis during early childhood; NTBC treatment is known to prolong HT1 children's lives-although liver fibrosis, cirrhosis, hepatocarcinoma, and corneal opacities sometimes occur. The mouse data in the present study are consistent with the possibility that endogenous oxidative stress-induced apoptosis may be the underlying cause of liver pathology seen in NTBC-treated HT1 patients. PMID- 12899939 TI - Hepatoprotective mechanism of schisandrin B: role of mitochondrial glutathione antioxidant status and heat shock proteins. AB - In this study, the time course of schisandrin B- (Sch B-) induced changes in hepatic mitochondrial glutathione antioxidant status (mtGAS) and heat shock protein (HSP) 25/70 induction was examined to study their differential roles in the hepatoprotection afforded by Sch B pretreatment against carbon tetrachloride (CCl(4)) toxicity in mice. Dimethyl diphenyl bicarboxylate (DDB), a nonhepatoprotective analog of Sch B, was also included for comparison. The results indicate that Sch B treatment (2 mmol/kg) produced maximum enhancement in hepatic mtGAS and increases in both hepatic HSP 25 and HSP 70 levels at 24 h after dosing. While the extent of hepatoprotection afforded by Sch B pretreatment against CCl(4) was found to correlate inversely with the elapsed time postdosing, the protective effect was associated with the ability to sustain mtGAS and/or HSP 70 levels in a CCl(4)-intoxicated condition. On the other hand, DDB (2 mmol/kg) treatment, which did not sustain mtGAS and HSP 70 level, could not protect against CCl(4) toxicity. Abolition of the Sch B-mediated enhancement of mtGAS by buthionine sulfoximine/phorone did not completely abrogate the hepatoprotective action of Sch B. The results indicate that Sch B pretreatment independently enhances mtGAS and induces HSP 25/70 production, particularly under conditions of oxidative stress, thereby protecting against CCl(4) hepatotoxicity. PMID- 12899940 TI - Nitric oxide and cGMP activate the Ras-MAP kinase pathway-stimulating protein tyrosine phosphorylation in rabbit aortic endothelial cells. AB - The free radical nitric oxide is a very effective signal transducer, stimulating the enzyme guanylyl cyclase, the oncoprotein p21Ras, and protein tyrosine phosphorylation. In the present study using rabbit aortic endothelial cells (RAEC), it is demonstrated that the nitric-oxide-generating substances sodium nitroprusside and S-nitroso-N-acetylpenicillamine, and a stable analog of cyclic GMP, 8BrcGMP stimulate p21Ras activity. Tyrosine phosphorylation of cytosolic proteins was stimulated and intracellular production of cGMP was increased, indicating that the NO/cGMP-stimulated tyrosine phosphorylation-dependent signaling pathway is most likely associated with the activation of p21Ras. NO and cGMP-dependent activation of p21Ras result in binding of the oncoprotein to the Ras-binding domain of Raf-1 kinase. Incubation of RAEC with FPT II, a potent and selective inhibitor of p21Ras, prevented NO-dependent tyrosine phosphorylation. ODQ, a potent inhibitor of the soluble form of guanylyl cyclase, inhibited the signal as well. Conversely, the use of KT5823, a cGMP-dependent protein kinase (PKG) blocker, showed no effect on protein tyrosine phosphorylation. To further establish a role for p21Ras on the NO-stimulated tyrosine phosphorylation signaling pathway, RAEC were constitutively transfected with a dominant negative mutant of p21Ras, N17Ras. NO and cGMP-stimulated tyrosine phosphorylation were prevented in N17Ras-expressing RAEC exposed to NO donors and 8BrcGMP. The above findings indicate that NO and cGMP stimulation of protein tyrosine phosphorylation requires the participation of fully functional p21Ras. ERK1/2 MAP kinases and their subsequent targets, the transcription factors, lie downstream to Ras, Raf-1 kinase, and MEK. Treatment of both RAEC and mock-transfected RAEC with NO resulted in phosphorylation and activation of ERK1/2. On the other hand, NO did not stimulate phosphorylation of ERK1/2 in N17Ras-expressing RAEC. In addition, PD98059, a MEK inhibitor, prevented overall tyrosine phosphorylation and phosphorylation of ERK1/2. Upstream to Ras ERK1/2 MAP kinases target the EGF receptor. Incubation of RAEC or mock-transfected RAEC with NO donors resulted in activation of the EGF receptor autophosphorylation. PD98059 effectively blocked this activation. EGF receptor autophosphorylation was insensitive to NO stimulation in N17Ras-expressing RAEC. It is concluded that NO and cGMP stimulate a signaling pathway involving p21Ras-Raf-1 kinase-MEK-ERK1/2. Activation of this signaling pathway is connected to NO-stimulated overall tyrosine phosphorylation that also involves the transactivation of the EGF receptor mediated by ERK1/2. PMID- 12899941 TI - Modulation of hOGG1 DNA repair enzyme in human cultured cells in response to pro oxidant and antioxidant challenge. AB - The putative modulation of the base excision repair enzyme, human 8-oxoguanine glycosylase (hOGG1), important in the removal of the potentially mutagenic lesion 8-oxo-2'-deoxyguanosine (8-oxodG), was investigated in human cell culture models. The expression of specific mRNA and protein was measured following pro-oxidant and antioxidant treatments in one human lymphoblastoid and one keratinocyte line. The measurement of intracellular reactive oxygen species generation was monitored by a fluorogenic assay and potential genotoxic effects confirmed by the dose dependent increase in formamidopyrimidine-DNA glycosylase (Fpg) sensitive sites by alkaline unwinding following sub-lethal doses of hydrogen peroxide. The generation of a potentially antioxidant environment was assessed by the intracellular increase and extracellular depletion in ascorbic acid, confirmed by capillary electrophoresis. Despite these pro-oxidant and antioxidant treatments no significant change in mRNA of hOGG1 was observed in either cell line. Western analysis revealed that relatively high, yet noncytotoxic, doses of hydrogen peroxide caused a consistent approximate 50% decrease in hOGG1 protein in lymphoblastoid cells. The lack of upregulation of hOGG1 suggests the gene is constitutively expressed, which is further supported by studies examining the sequence of its promoter region. However, hOGG1 protein turnover may be sensitive to intracellular redox changes. PMID- 12899942 TI - T cell receptor-stimulated generation of hydrogen peroxide inhibits MEK-ERK activation and lck serine phosphorylation. AB - Previous studies indicated that antigen receptor (TcR) stimulation of mature T cells induced rapid generation of reactive oxygen species (ROS). The goal of the current study was to examine the role(s) of ROS in TcR signal transduction, with a focus upon the redox-sensitive MAPK family. TcR cross-linking of primary human T blasts and Jurkat human T cells rapidly activated the ERK, JNK, p38 and Akt kinases within minutes, and was temporally associated with TcR-stimulated production of hydrogen peroxide (H(2)O(2)). TcR-induced activation of ERK was selectively augmented and sustained in the presence of pharmacologic antioxidants that can quench or inhibit H(2)O(2) production (NAC, MnTBAP and Ebselen, but not DPI), while activation of JNK and Akt were largely unaffected. This was paralleled by concurrent changes in MEK1/2 phosphorylation, suggesting that ROS acted upstream of MEK-ERK activation. Molecular targeting of H(2)O(2) by overexpression of peroxiredoxin II, a thioredoxin dependent peroxidase, also increased and sustained ERK and MEK activation upon TcR cross-linking. Enhancement of ERK phosphorylation by antioxidants correlated with increased and sustained serine phosphorylation of the src-family kinase lck, a known ERK substrate. Thus, the data suggest that TcR-stimulated production of hydrogen peroxide negatively feeds back to dampen antigen-stimulated ERK activation and this redox-dependent regulation may serve to modulate key steps in TcR signaling. PMID- 12899943 TI - Copper blocks quinolinic acid neurotoxicity in rats: contribution of antioxidant systems. AB - Reactive oxygen species and oxidative stress are involved in quinolinic acid (QUIN)-induced neurotoxicity. QUIN, a N-methyl-D-aspartate receptor (NMDAr) agonist and prooxidant molecule, produces NMDAr overactivation, excitotoxic events, and direct reactive oxygen species formation. Copper is an essential metal exhibiting both modulatory effects on neuronal excitatory activity and antioxidant properties. To investigate whether this metal is able to counteract the neurotoxic and oxidative actions of QUIN, we administered copper (as CuSO(4)) intraperitoneally to rats (2.5, 5.0, 7.5, and 10.0 mg/kg) 30 min before the striatal infusion of 1 microliter of QUIN (240 nmol). A 5.0 mg/kg CuSO(4) dose significantly increased the copper content in the striatum, reduced the neurotoxicity measured both as circling behavior and striatal gamma-aminobutyric acid (GABA) depletion, and blocked the oxidative injury evaluated as striatal lipid peroxidation (LP). In addition, copper reduced the QUIN-induced decreased striatal activity of Cu,Zn-dependent superoxide dismutase, and increased the ferroxidase activity of ceruloplasmin in cerebrospinal fluid from QUIN-treated rats. However, copper also produced significant increases of plasma lactate dehydrogenase activity and mortality at the highest doses employed (7.5 and 10.0 mg/kg). These results show that at low doses, copper exerts a protective effect on in vivo QUIN neurotoxicity. PMID- 12899944 TI - Effect of prenatal exposure to ethanol on hepatic elongation factor-2 and proteome in 21 d old rats: protective effect of folic acid. AB - In this article, we study the effects of ethanol intake during pregnancy and lactation on hepatic and pancreatic elongation factor-2 (EF-2) of 21 d old progeny. At the same time, the effect of ethanol on the level of other relevant hepatic proteins was determined using proteomic analysis. The results show that ethanol not only produces a general increase of protein oxidation, but also produces an important depletion of EF-2 and several other proteins. Among the hepatic proteins affected by ethanol, the concomitant supplementation with folic acid to alcoholic mother rats prevented EF-2, RhoGDI-1, ER-60 protease, and gelsolin depletion. This protective effect of folic acid may be related to its antioxidant properties and suggests that this vitamin may be useful in minimizing the effect of ethanol in the uterus and lactation exposure of the progeny. PMID- 12899945 TI - Relief of oxidative stress by ascorbic acid delays cellular senescence of normal human and Werner syndrome fibroblast cells. AB - Primary human cells have a definite life span and enter into cellular senescence before ceasing cell growth. Oxidative stress produced by aerobic metabolism has been shown to accelerate cellular senescence. Here, we demonstrated that ascorbic acid, used as an antioxygenic reagent, delayed cellular senescence in a continuous culture of normal human embryonic cells, human adult skin fibroblast cells, and Werner syndrome (WS) cells. The results using human embryonic cells showed that treatment with ascorbic acid phospholic ester magnesium salt (APM) decreased the level of oxidative stress, and extended the replicative life span. The effect of APM to extend the replicative life span was also shown in normal human adult cells and WS cells. To understand the mechanism of extension of cellular life span, we determined the telomere lengths of human embryonic cells, both with and without APM treatment, and demonstrated that APM treatment reduced the rate of telomere shortening. The present results indicate that constitutive oxidative stress plays a role in determining the replicative life span and that suppression of oxidative stress by an antioxidative agent, APM, extends the replicative life span by reducing the rate of telomere shortening. PMID- 12899946 TI - EPR spectroscopy studies on the structural transition of nitrosyl hemoglobin in the arterial-venous cycle of DEANO-treated rats as it relates to the proposed nitrosyl hemoglobin/nitrosothiol hemoglobin exchange. AB - In vivo studies show a dynamic cycle in which alpha-nitrosylated hemoglobin is mainly in the relaxed state in arterial blood of rats treated with 2-(N,N diethylamino)-diazenolate-2-oxide, but converts mainly to the tense state during the arterial-venous transit. A detailed analysis shows that different electron paramagnetic resonance spectra recorded for alpha-nitrosyl hemoglobin in arterial and venous blood at 77 K originate only from a different ratio between 5- and 6 coordinate heme without any change in the concentration of nitrosyl hemoglobin. In venous blood, the five- and six-coordination equilibrium of the alpha-nitrosyl heme is shifted in favor of the 5-coordinate state (58% venous vs. 20% arterial). These results are not consistent with the recently proposed exchange of nitrosyl heme with the beta-93 nitrosothiol group of hemoglobin during the arterial-venous cycle. PMID- 12899947 TI - Stir bar sorptive extraction-thermal desorption-capillary GC-MS for profiling and target component analysis of pharmaceutical drugs in urine. AB - Stir bar sorptive extraction (SBSE) in combination with thermal desorption (TD) on-line coupled to capillary gas chromatography-mass spectrometry (CGC-MS) was applied to the analysis of pharmaceutical drug compounds and metabolites in urine. SBSE implies stirring of the aqueous sample (urine, blood, etc.) with a glass stir bar coated with a thick layer (24 microl) of polydimethylsiloxane (PDMS) for sorptive enrichment of the analytes of interest. In combination with quantitative TD, on-line coupled with CGC-MS, the technique showed to be very versatile and sensitive for the analysis of a wide range of drug substances. Moreover, the relative high enrichment efficiencies of SBSE allow to use mass spectrometric detection (MSD) in the full scan mode. In situ derivatization of polar compounds before SBSE is demonstrated for the analysis of paracetamol and this resulted in both improved chromatographic behavior and higher sensitivity. The quantitative performance of SBSE-TD-CGC-MS is illustrated with the analysis of some barbiturates in urine. PMID- 12899948 TI - Characterization of antisense oligonucleotide-peptide conjugates with negative ionization electrospray mass spectrometry and liquid chromatography-mass spectrometry. AB - Covalent post-synthesis or solid-phase conjugation of peptides to oligonucleotides has been reported as a possible method of delivering antisense oligonucleotides into cells. While synthesis strategies for preparing these conjugates have been widely addressed, few detailed reports on their structural characterization have been published. This paper discusses the negative ion electrospray ionization mass spectrometric (ESI-MS) and liquid chromatography mass spectrometric (LC-MS) analysis of various peptide-oligonucleotide conjugates ranging from small T(6)-nucleopeptides to large peptide-oligonucleotide phosphorothioate conjugates and ribozyme-peptide hybrids (3-13 kDa). Molecular weight determination with mass errors of 0.1-3.1 amu were conducted, employing on line IP-RP-HPLC and high m/z range mode to facilitate the analysis of large compounds and difficult modifications. PMID- 12899949 TI - Quantitative determination of paclitaxel in human plasma using semi-automated liquid-liquid extraction in conjunction with liquid chromatography/tandem mass spectrometry. AB - This paper describes a high-throughput sample preparation procedure combined with LC-MS/MS analysis to measure paclitaxel in human plasma. Paclitaxel and an internal standard were extracted from plasma by a semi-automated robotic method using liquid-liquid extraction. Thereafter compounds were separated on a RP C18 column. Detection was by a PE Sciex API 3000 mass spectrometer equipped with a TurboIonSpray interface. The compounds were detected in positive ion mode using the mass transition m/z 854.6-->286.2 and m/z 831.6-->263.2 for paclitaxel and the internal standard, respectively. The limit of quantitation for paclitaxel was 1 ng/ml with an imprecision of 5.2% following extraction of 0.1 ml of plasma. Linearity was confirmed over the whole calibration range (1-1000 ng/ml) with correlation coefficients higher than 0.99 indicating good fits of the regression models. The inter and intra-day precision was better than 9.5% and the accuracy ranged from 90.3 to 104.4%. The assay was simple, fast, specific and exhibited excellent ruggedness. PMID- 12899950 TI - LC-MS-MS determination of brostallicin in human plasma following automated on line SPE. AB - LC-MS-MS method using automated on-line solid-phase extraction (SPE) has been developed and validated for the quantitation of brostallicin (I), a new distamycin derivative, in human plasma. I is a DNA minor groove binder currently under phase I-II clinical evaluation as an anticancer drug. Plasma (0.4 ml) was spiked with 0.2 ml stable label IS solution and placed in a 96-well plate maintained at +4 degrees C. Aliquots of 0.1 ml of prepared samples were loaded into the on-line SPE HySphere Resin SH cartridges (10 mm x 2 mm ID) and the analytes back eluted with the mobile phase into LC-MS-MS system. A Platinum Cyano column (100 mm x 4.6 mm, 3.6 microm) was used to perform the chromatographic analysis. The mobile phase was acetonitrile-ammonium formate buffer (pH 3.5; 20 mM) (70:30, v/v) at a flow rate of 1.0 ml/min. LC flow was split so that 300 microl/min was directed toward the mass spectrometer interface. Retention time of I was 2.6 min and the total cycle time was 8 min. MS detection used an Applied Biosystems/MDS SCIEX API 365 with a TurboIonSpray interface and MRM (m/Z: 725/257 for I and m/Z: 729/257 for IS) operated in positive ion mode. The method was validated over the calibration range 0.124-497 ng/ml. A negligible carry-over effect from the system was observed. In spite of the known instability of I in human plasma (about 20% decrease over 12 h), the ratio analyte/IS peak area showed good stability over the analysis time required for 96 samples. The automated on-line SPE method can be considered as a valid alternative to the off line manual SPE procedure previously developed. PMID- 12899951 TI - A sensitive LC/MS/MS method using silica column and aqueous-organic mobile phase for the analysis of loratadine and descarboethoxy-loratadine in human plasma. AB - A sensitive method using liquid chromatography with tandem mass spectrometric detection (LC/MS/MS) was developed and validated for the simultaneous analysis of antihistamine drug loratadine (LOR) and its active metabolite descarboethoxy loratadine (DCL) in human plasma. Deuterated analytes, i.e. LOR-d(3) and DCL-d(3) were used as the internal standards (I.S.). Analytes were extracted from alkalized human plasma by liquid/liquid extraction using hexane. The extract was evaporated to dryness under nitrogen, reconstituted with 0.1% (v/v) of trifluoroacetic acid (TFA) in acetonitrile, and injected onto a 50 x 3.0 mm I.D. 5 microm, silica column with an aqueous-organic mobile phase consisted of acetonitrile, water, and TFA (90:10:0.1, v/v/v). The chromatographic run time was 3.0 min per injection and flow rate was 0.5 ml/min. The retention time was 1.2 and 2.0 min for LOR and DCL, respectively. The tandem mass spectrometric detection was by monitoring singly charged precursor-->product ion transitions: 383-->337 (m/z) for LOR, 311-->259 (m/z) for DCL, 388-->342 (m/z) for LOR-d(3), and 316-->262 (m/z) for DCL-d(3). The low limit of quantitation (LLOQ) was 10 pg/ml for LOR and 25 pg/ml for DCL. The inter-day precision of the quality control (QC) samples was 3.5-9.4% relative standard deviation (R.S.D.). The inter day accuracy of the QC samples was 99.0-107.9% of the nominal values. PMID- 12899952 TI - Sensitive determination of buprenorphine and its N-dealkylated metabolite norbuprenorphine in human plasma by liquid chromatography coupled to tandem mass spectrometry. AB - A highly sensitive method based on liquid chromatography-tandem mass spectrometry (LC-MS/MS) has been developed for the quantitative determination of buprenorphine and its active metabolite norbuprenorphine in human plasma. Automated solid phase extraction (SPE) on disposable extraction cartridges (DEC) is used to isolate the compounds from the biological matrix and to prepare a cleaner sample before injection and analysis in the LC-MS/MS system. After conditioning, the plasma sample (1.0 ml) is loaded on the DEC filled with octyl silica (C8) and washed with water. The analytes are, therefore, eluted by dispensing methanol containing 0.1% of acetic acid. The eluate is collected and evaporated to dryness. The residue is dissolved in mobile phase and an aliquot is injected in the LC-MS/MS system. On-line LC-MS/MS system using atmospheric pressure chemical ionization (APCI) has been developed for the determination of buprenorphine and norbuprenorphine. The separation is obtained on a RP-18 stationary phase using a mobile phase consisting in a mixture of methanol and 50 mM ammonium acetate solution (50:50, v/v). Clonazepam is used as internal standard (IS). The MS/MS ion transitions monitored are m/z 468-->468, 414-->414 and 316-->270 for buprenorphine, norbuprenorphine and clonazepam, respectively. The method was validated regarding recovery, linearity, precision and accuracy. The limits of quantification (LOQs) were around 10 pg/ml for buprenorphine and 50 pg/ml for norbuprenorphine. PMID- 12899953 TI - Liquid chromatography/atmospheric pressure chemical ionization ion trap mass spectrometry of terpene lactones in plasma of animals. AB - Liquid chromatography/atmospheric pressure chemical ionization ion trap mass spectrometry (LC/APCI-ITMS) was applied to evaluate the bioavailibility of two different forms (free and complexed with soy phospholipids) of pure bilobalide and ginkgolide B in rats after acute administration. The same technique was used to measure the levels of ginkgolide A, B and bilobalide in plasma of guinea pigs fed Ginkgo biloba extract enriched in terpene lactones after chronic administration. The ratio R(P)/R(A) increased two to four times after the administration in the phytosomic form, where R(P) and R(A) represent the percentage ratio between the concentration of each terpene lactone in plasma and in the administrated form, respectively. PMID- 12899954 TI - Comparative biotransformation and disposition studies of nabumetone in humans and minipigs using high-performance liquid chromatography with ultraviolet, fluorescence and mass spectrometric detection. AB - The disposition of the non-steroidal anti-inflammatory drug (NSAID) nabumetone after a single oral dose administration of nabumetone tablets to humans and minipigs was investigated. Nabumetone is a prodrug, which is metabolized in the organism to the principal pharmacodynamically active metabolite -- 6-methoxy-2 naphthylacetic acid (6-MNA), and some other minor metabolites (carbonyl group reduction products, O-desmethylation products and their conjugates with glucuronic and sulphuric acids). Standards of the above-mentioned metabolites were prepared using simple synthetic procedures and their structures were confirmed by NMR and mass spectrometry. A simple HPLC method for the simultaneous determination of nabumetone, 6-MNA and the other metabolites was developed, validated and used for xenobiochemical and pharmacokinetic studies in humans and minipigs and for distribution studies in minipigs. Naproxen was chosen as the internal standard (I.S.), both UV (for higher concentrations) and fluorescence detection (for very low concentrations) were used. The identity of the nabumetone metabolites in biological samples was confirmed using HPLC-MS experiments. Pharmacokinetics of nabumetone, 6-MNA and 6-HNA (6-hydroxy-2-naphthylacetic acid) in human and minipig plasma was evaluated and compared. The concentration levels of nabumetone metabolites in urine, bile and synovial fluid were also evaluated. PMID- 12899955 TI - LC-APCI-MS/MS analysis of urinary 8-hydroxy-2'-deoxyguanosine. AB - 8-Hydroxy-2'-deoxyguanosine (8OHdG) is regarded as an important biomarker of oxidative DNA damage and it may be estimated by using different techniques in various biological matrices, most notably DNA and urine. In the case of DNA, artifactual oxidation may take place during the isolation of DNA, its hydrolysis and possible derivatization (as for GC-MS), invalidating the measurement of 8OHdG. Therefore, the direct analysis of 8OHdG excreted into urine was preferred. Interferences from the urine matrix were excluded by applying LC-APCI-MS/MS in the multiple reaction monitoring (MRM) mode. The abundant fragment ion at m/z 168 arising from 8OHdG was monitored in the urine sample of volunteers supplemented with tomato concentrate for different times. The procedure allowed the detection of levels of 8OHdG as low as 1 ng/ml in urine sample. PMID- 12899956 TI - RP-HPLC separation of the diastereomers of technetium-99m labelled tropanes and identity confirmation using radio-LC-MS. AB - 99mTc-TRODAT-1 (technetium(V)-oxo-2-[[2-[[[3-(4-chlorophenyl)-8-methyl-8 azabicyclo[3.2.1]oct-2-yl]methyl](2-mercaptoethyl)amino]ethyl]]amino] ethanethiolato(3-)) and 99mTc-TRODAT-M, the 4-methylphenyl derivative of 99mTc TRODAT-1, are at this moment being evaluated in clinical trials as imaging agents for the central nervous dopamine transporter system. Both compounds are formed as a mixture of two major diastereomers. As the tracer concentration in preparations for clinical investigations is very low (30-150 pmol/ml), identification of these 99mTc-complexes was, up to now, carried out indirectly using X-ray diffraction analysis of the corresponding rhenium complexes which can be synthesized in gram amounts. In this study, we developed a convenient and practical reversed phase HPLC-method for purification and isolation of the respective diastereomers of 99mTc-TRODAT-1 and three of its derivatives using mixtures of solvents which are compatible with biological studies, i.e. aqueous buffers and ethanol. Furthermore, direct identity confirmation of the 99mTc-complexes using radio-LC MS was successfully elaborated. PMID- 12899957 TI - Identity confirmation of 99mTc-MAG3, 99mTc-sestamibi and 99mTc-ECD using radio-LC MS. AB - Due to the low concentrations in which 99mTc-radiopharmaceuticals are obtained (4 40 ng/ml), confirmation of the identity of these tracer agents in the European Pharmacopoeia is generally performed only indirectly by assessment of their retention times on RP-HPLC. We have investigated whether it is possible to obtain more direct proof of the identity of technetium-99m labelled radiopharmaceuticals using radio-LC-MS. As representative examples, negatively charged 99mTc-MAG3, positively charged 99mTc-Sestamibi and neutral 99mTc-ECD were used. The three technetium-99m radiopharmaceuticals were prepared in several conditions to obtain variable relative amounts of radiochemical impurities and variable concentrations of the complexes (pico- to nanomolar). The preparations were analyzed on a reversed phase C18 HPLC column using a radio-LC-MS system equipped with a time of flight mass spectrometer with electrospray ionization in positive (99mTc Sestamibi, 99mTc-ECD) or negative (99mTc-MAG3, 99mTc-ECD) mode. For each of the studied complexes, the main peak in the radiometric channel coincided with the expected molecular ion mass of the corresponding technetium complex in the mass spectrometer channel. The relative error on the measured accurate mass was in the range of 10 ppm. The identity of several radiochemical impurities of the three technetium complexes was also confirmed or established. It is concluded that radio-LC-MS can be a sensitive aid in quality control of 'no carrier added' radiopharmaceuticals. PMID- 12899958 TI - An efficient HPLC method for the analysis of isomeric purity of technetium-99m exametazime and identity confirmation using LC-MS. AB - 99mTc-exametazime (99mTc-d,l-HMPAO, 99mTc-d,l-hexamethylpropyleneamine oxime) is a neutral rather unstable complex of short-lived 99mTc (t(1/2)=6 h) with the d,l isomer (mixture of D,D- and L,L-isomers) of a bis-amine bis-oxime tetraligand. It is widely used for measurement of regional cerebral perfusion in nuclear medicine. The meso-isomer (D,L-form) should not be present in a preparation as it is not retained in brain and thus does not provide clinically useful information. Meso-HMPAO is removed from the ligand during the synthesis procedure by repeated recrystallization, but can still be present as impurity in d,l-isomer. Due to the lack of a suitable chromatographic method for analysis of the isomeric purity of 99mTc-exametazime preparations, United States Pharmacopoeia 25 (USP 25) prescribes a biological test in rats for quality control purpose. In this study, we developed a suitable high-performance liquid chromatography (HPLC) method which allows to demonstrate the relative amounts of d,l- and meso-isomer in 99mTc exametazime and so obviates the need for a biodistribution test in animals as part of the quality control. Due to the low concentrations in which 99mTc-d,l HMPAO is obtained (typically 2-6 ng/ml), confirmation of the identity of 99mTc d,l-HMPAO in the monograph of the European Pharmacopoeia is now performed only indirectly by TLC and assessment of its retention time on RP-HPLC. To investigate the potential of radio-LC-MS for assessment of the identity of 99mTc-exametazime, 99mTc-d,l-HMPAO and 99mTc-meso-HMPAO prepared using a Tc-rich eluate were analyzed using a radio-LC-MS system equipped with a time-of-flight mass spectrometer with electrospray ionization. The main peak in the radiometric channel coincided with the molecular ion mass of 99mTc-d,l-HMPAO in the mass spectrometer channel and the measured accurate mass differed only by 0.26 ppm from the theoretical mass. The identity of 99mTc-meso-HMPAO was also confirmed. Thus, radio-LC-MS allowed to obtain strong evidence for the structure of 99mTc d,l-HMPAO and 99mTc-meso-HMPAO at nanomolar concentration. It is concluded that radio-LC-MS can become a sensitive aid in quality control of "no carrier added" radiopharmaceutical preparations. PMID- 12899959 TI - Polypyrrole based DNA hybridization assays: study of label free detection processes versus fluorescence on microchips. AB - In this paper, we present different ways to detect DNA hybridization on a solid support. The grafting chemistry is based on the electro-controlled copolymerization of a pyrrole-modified oligonucleotide and pyrrole. This process allows an easy functionalization of conducting materials. Three kind of devices were studied: silicon chips bearing an array of addressable 50 or 4 microm microelectrodes, quartz crystal microbalance (QCM) and a non patterned gold/glass slide bearing 500 microm spots. Each device is compatible with a specific detection process: a classical indirect fluorescence detection for the microchips, a microgravimetric measurement for the QCM and a surface plasmon resonance imaging process (SPRi) for the gold slides. Both QCM and SPRi are a label-free real time detection process whereas the fluorescence methodology gives end-point data but only the fluorescence and the SPRi give multiparametric results. Although the hybridization experiments show that the detection limit for an oligonucleotide is better for the fluorescence (1-10 pM) than that found for SPRi (10 nM) and QCM (250 nM), the information content of real time measurement techniques such as SPRi is of interest for many biological studies. PMID- 12899960 TI - Optical biosensor analysis in studying herpes simplex virus glycoprotein D binding to target nectin1 receptor. AB - Studies on molecular interactions between cellular receptors of herpes simplex virus (HSV) and the viral glycoproteins showing receptor-binding activity are of great relevance for understanding the molecular basis of virus entry. Information on such interactions further provides the basis for a rational design of antiviral drugs. A variety of biochemical and biophysical methodologies are used for determining the binding parameters of interacting biomolecules. Most of them require relatively high amounts of the analyzed compounds, or the use of labeled target molecules. Here, we report the study of the binding of two recombinant forms of HSV glycoprotein D, gD(Delta 290-299t) and gD(305t), and a recombinant form of the human cellular receptor for HSV, nectin1-Fc, by using an optical biosensor (IAsys Plus, Affinity Sensors, UK). This device detects and quantifies the changes in refractive index in the vicinity of the surface of sensor chips to which ligands are immobilized. The changes in the refractive index are proportional to the change in the absorbed mass, thus the analysis allows the monitoring of the interaction process in real-time and the determination of the binding parameters. HSV cellular receptor has been immobilized on the surface of the biosensor cuvette, bearing a carboxymethyl dextran layer. The immobilized receptor cuvette was then used for the binding experiments of the two glycoproteins. A significant difference in their dissociation constants was determined, showing for the gD(Delta 290-299t) protein a much higher affinity (K(D), 2.8 x 10(-7) M) with respect to gD(305t) (K(D), 2.8 x 10(-6) M). The active ligand concentration decreased on time, however the binding properties of the immobilized receptor were maintained over 5 weeks. PMID- 12899961 TI - Rapid screening of small ligand affinity to human serum albumin by an optical biosensor. AB - Here we report the use of IAsys biosensor technology for determining the binding parameters of low molecular weight compounds, such as warfarin and bilirubin, to surface immobilized human serum albumin. The protein was covalently immobilized on the surface of the biosensor cuvette, bearing a carboxymethyl dextran layer, through a condensing reaction between the carboxyl groups of the biosensor surface and epsilon-amine groups of protein lysine residues. This system detects and quantifies the changes in refractive index in the vicinity of the surface of the sensor chip to which the protein is immobilized. The changes in the refractive index are proportional to the change in the absorbed mass, thus the analysis allows the monitoring of the interaction process and the determination of the binding parameters. Optical biosensor analysis, most suited for studying protein/protein or protein/nucleic acid interactions, was sensitive enough to monitor the binding of low molecular weight compounds to human serum albumin and then suitable for a rapid screening of libraries of potential drugs when bioavailability is the research target. PMID- 12899962 TI - Lipases for biocatalysis: development of a chromatographic bioreactor. AB - The development of a new chromatographic reactor based on immobilized Candida rugosa lipase (CRL) is described. The chromatographic system has been used to evaluate the rate differences by which the product enantiomers of esterolytic reactions catalyzed by immobilized CRL are obtained. The method has been applied to a series of racemic 2-aryloxyalkanoic acids and isosteric analogous methyl esters and to some non-steroidal antiinflammatory drugs 2-arylpropanoic acids methyl esters in order to study the structure effects on reaction rate and enantioselectivity. Lipase from C. rugosa has been non-covalently and covalently immobilized on HPLC chromatographic silica supports to develop an immobilized enzyme reactor (IMER). The reactor was connected through a switching valve to an analytical reversed-phase column, which was used for the on-line determination of the hydrolysis rate by peak area integration. The enantiomeric excess of the hydrolytic reaction products was determined off-line on a CSP utilizing immobilized penicillin G acylase (PGA-CSP). PMID- 12899963 TI - Determination of the antioxidant capacity of samples of different types of tea, or of beverages based on tea or other herbal products, using a superoxide dismutase biosensor. AB - Research was performed to experimentally evaluate the antioxidant capacity of different plant products sold by herbalists (ginger, dog rose, ginseng and camomile) and of several types of tea (ordinary tea, green tea, detheinated tea, lemon and peach flavoured tea) using a superoxide dismutase (SOD) biosensor recently developed by the present authors. Measurements were carried out by comparing biosensor response to the superoxide radical produced in solution using the xanthine-xanthine oxidase system, both in the presence and absence of the antioxidant sample considered. Precision of antioxidant capacity measures for herbal products and for non diluted samples was good, generally with a R.S.D.%< or =10% and a LOD value about 0.1 for relative antioxidant capacity. Also a "pool" of polyphenols from different tea samples was measured using a tyrosinase biosensor (LOD approximately 2 microM). PMID- 12899964 TI - Determination of hydrogen peroxide in disinfectant solutions using a biosensor with two antagonist enzymes. AB - The development and characterisation of a new biosensor for hydroperoxides is described, which is obtained by combining an oxygen gas diffusion amperometric electrode and two immobilized enzymes (peroxidase and tyrosinase) working in parallel and competing for the same substrate (catechol). The response of the biosensor to several hydroperoxides was investigated (LOD=0.5.10(-4) M for hydrogen peroxide). It was experimentally found that the biosensor is able to respond also to aqueous solutions of ionic peroxides (LOD=0.2.10(-4) M for potassium peroxidisulphate). The biosensor was applied to the determination of the hydrogen peroxide content of pharmaceutical products, i.e. aqueous disinfectant solutions (RSD% < or =0.5; recoveries by standard addition method between 96.0 and 98.5%). PMID- 12899965 TI - An analysis of the SFSTP guide on validation of chromatographic bioanalytical methods: progress and limitations. AB - The Societe Francaise des Sciences et Techniques Pharmaceutiques (SFSTP) published in 1997 a guide on the validation of chromatographic bio-analytical methods, which introduces new concepts in three different areas: stages of the validation, test of acceptability of a method and design of experiments to perform. In 'stages of validation', the SFSTP guide requires two phases to validate a method. The first phase, called 'prevalidation', is intended to (1) identify the model to use for the calibration curve; (2) evaluate the limits of quantitation; and (3) provide good estimates of the precision and bias of the method before designing the 'validation' phase per se. In the 'test of acceptability', the use of the interval hypotheses is envisaged by the SFSTP guide, not on the parameters of bias and precision, but on individual results by mixing mean bias and intermediate precision in a single test. The SFSTP guide also avoids the use of Satterthwaite's df for testing the acceptability. The reasons for those choices are discussed extensively. In 'design of experiments', much effort has been devoted to improving the quality of results by optimally designing and sizing the experiments to perform in validation. The rationale for using near D-optimal designs for the calibration curve is demonstrated and sample sizes are proposed to correctly size the validation experiments. PMID- 12899966 TI - Measurement uncertainty from validation and duplicate analysis results in HPLC analysis of multivitamin preparations and nutrients with different galenic forms. AB - An approach to calculate the measurement uncertainty in the HPLC analysis of several hydro- and liposoluble vitamins in multivitamin preparations with different galenic composition and properties is described. In the first instance it is examined if duplicate analysis results, obtained with a fully validated analysis method on different lots of an effervescent tablet preparation spread over several points of time, might contribute to calculate the measurement uncertainty of the HPLC method used and if the established uncertainty is acceptable in the assessment of compliance with the legal content limits. Analysis of variance (ANOVA) and precision calculations, based on the ISO 5725-2 norm are applied on the analysis results obtained to estimate precision components, necessary to derive the measurement uncertainty. In the second instance it is demonstrated to which extent the fully validated method of analysis for effervescent tablets is applicable to other galenic forms as e.g. capsules with oily emulsions, tablets, coated tablets, oral solutions, em leader and which specific modifications in the analysis steps are involved. By means of duplicate analysis results, acquired from a large series of real samples over a considerable period of time and classified according to their similarity in content, galenic forms and matrices, estimations of measurement uncertainty calculations are shown. PMID- 12899967 TI - Application of capillary electrophoresis with pH-mediated sample stacking to analysis of coumarin metabolites in microsomal incubations. AB - A sensitive method for the analysis of metabolites of coumarin by capillary electrophoresis (CE), incorporating pH-mediated sample stacking, was developed. The analytes were detected in phosphate buffer (pH 7.5; 25 mM), the matrix of the microsomal incubations. Detection was by direct UV absorbance. The three metabolites studied were 7-hydroxycoumarin (7-OHC), 4-hydroxycoumarin (4-OHC) and 2-hydroxyphenylacetic acid (HPAA), and the limits of detection of the analytes were 0.1, 0.5 and 0.3 microM, respectively. The developed method was then applied to microsomal incubations of coumarin. Male Cynomologus monkey microsomes were used in the study and 7-OHC was detected in the incubation mixture. PMID- 12899968 TI - Capillary electrophoresis: a tool for studying interactions of glycans/proteoglycans with growth factors. AB - Heparan sulfate (HS) and heparin bind to various growth factors and modulate their activities. Interactions of heparin and HS with members of the fibroblast growth factor (FGF) family are prerequisites for binding of FGFs to their high affinity cell receptors. The sulfation patterns of distinct oligosaccharide domains within heparin and HS chains determine their high affinity binding with basic FGF (bFGF). In order to study the structural basis of interactions of HS with bFGF, we developed a capillary electrophoresis (CE) method in order to monitor the ability of HS-derived oligosaccharides to bind this growth factor. HS was degraded to Delta-di- and Delta-oligosaccharides with digestion with heparitinase and the obtained Delta-saccharides were analyzed by capillary zone electrophoresis (CZE), using 50 mM phosphate, pH 3.5, as operating buffer, reversed polarity (30 kV) and detection at 232 nm. Under these conditions all differently sulfated HS Delta-disaccharides and the various Delta-oligosaccharide groups were resolved. Following incubation of the digest with bFGF and re electrophoresis of the mixture, the bFGF interacting oligosaccharide groups were easily detected and identified. In view of the obtained results, CE is a multipotent analytical tool for determining disaccharide composition in HS, separating the various oligosaccharide groups produced by the action of heparitinase and identifying those interacting with bFGF. PMID- 12899969 TI - Fully automated LC method for the determination of sotalol in human plasma using restricted access material with cation exchange properties for sample clean-up. AB - A simple and rapid fully automated bio-analytical method for the liquid chromatographic (LC) determination of sotalol in human plasma has been described. The method is based on the use of a new kind of porous silica restricted access material (RAM) with cation exchange properties for sample clean-up. 100 microl of plasma samples were directly injected into the precolumn coupled on-line to a reversed-phase column (RP-Select B) by means of column switching system. The plasma matrix was washed out for 10 min using a washing liquid composed of 2 mM lithium perchlorate and methanol (97:3; v/v). By rotation of the switching valve, the analytes were then eluted in back-flush mode for 2 min and transferred to the analytical column by the LC mobile phase constituted of a mixture of methanol and 50 mM potassium phosphate buffer (pH 7.0) containing 1 mM 1-octanesulphonic acid sodium salt (20:80; v/v). The flow-rate was 1.0 ml/min and sotalol was detected using fluorescence detection at 235 and 300 nm as excitation and emission wavelengths, respectively. The method was then validated using a new approach based on accuracy profile over a concentration range from 5 to 500 ng/ml. The limit of quantitation (LOQ) was 5 ng/ml and the total analysis time was 19 min. PMID- 12899970 TI - Application of an alkyl-diol silica precolumn in a column-switching system for the determination of meloxicam in plasma. AB - The group of LiChrospher alkyl-diol silica (ADS) phases that make part of the unique family of restricted-access materials, have been developed as special packings used in the liquid chromatographic integrated sample processing of biofluids. The advantage of these phases lies in the possibility of direct injection of untreated plasma. An on-line elimination of the protein matrix is achieved with a quantitative recovery together with an on-column enrichment. The present method describes a hand-operated on-line switching high-performance liquid chromatographic system for the determination of meloxicam. Spiked plasma samples were introduced on the ADS precolumn using a 0.05 M phosphate buffer, pH 6.0. After washing with the buffer the ADS column was backflushed with the mobile phase 0.05 M phosphate buffer-30% (v/v) acetonitrile (ACN)-25 mM t-butylamine (TBA) at a pH of 7.0, thus transferring the analyte to the analytical column LiChrocart 125-4 LiChrospher RP-8. The eluent was monitored by a UV-detector set at 364 nm. The developed column-switching method is fully applicable to plasma injections. PMID- 12899971 TI - Experimental design approach for the optimisation of a HPLC-fluorimetric method for the quantitation of the angiotensin II receptor antagonist telmisartan in urine. AB - A high performance liquid chromatographic method with fluorimetric detection has been developed for the quantitation of the angiotensin II receptor antagonist (ARA II) 4-((2-n-propyl-4-methyl-6-(1-methylbenzimidazol-2-yl)-benzimidazol-1 yl)methyl)biphenyl-2-carboxylic acid (telmisartan) in urine, using a Novapak C18 column 3.9 x 150 mm, 4 microm. The mobile phase consisted of a mixture acetonitrile-phosphate buffer (pH 6.0, 5 mM) (45:55, v/v) pumped at a flow rate of 0.5 ml min(-1). Effluent was monitored at excitation and emission wavelengths of 305 and 365 nm, respectively. Separation was carried out at room temperature. Chromatographic variables were optimised by means of experimental design. A clean up step was used for urine samples consisting of a solid-phase extraction procedure with C8 cartridges and methanol as eluent. This method proved to be accurate (RE from -12 to 6%), precise (intra- and inter-day coefficients of variation (CV) were lower than 8%) and sensitive enough (limit of quantitation (LOQ), ca. 1 microg l(-1)) to be applied to the determination of the active drug in urine samples obtained from hypertensive patients. Concentration levels of telmisartan at different time intervals (from 0 up to 36 h after oral intake) were monitored. PMID- 12899972 TI - New HPLC method for separation of blood plasma phospholipids. AB - The aim of the present work was to develop a new HPLC method for separation of phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylinositol (PI) and lysophosphatidylcholine (LPC) from small-volume samples of blood plasma. Human plasma glycerophospholipids were separated by liquid-liquid extraction method followed by solid phase extraction (SPE) on aminopropyl columns. Reversed phase Sephasil C8 column (10 cm x 2.1 mm, I.D. 5 microm) and micropreparative chromatograph "SMART" were used for separation of PC, PE, LPC and PI from SPE phospholipids extract. Binary-step gradient of eluent A: acetonitrile-methanol (130:5, v/v) and B (0.01% trifluoroacetic acid) provided good, fast and reproducible resolution of investigated phospholipids classes in 12 min at 30 degrees C. Eluted phospholipids were detected at wavelengths lambda=235 and 254 nm. This method made it possible to determine quantitatively: 5 microg ml(-1) PC, 1 microg ml(-1) LPC, 4 microg ml(-1) PE and 3 microg ml(-1) PI in blood plasma samples. PMID- 12899973 TI - The study of codeine-gluthetimide pharmacokinetic interaction in rats. AB - A high-performance liquid chromatographic (HPLC) assay with native fluorescence detection was developed for the simultaneous quantification of codeine and its two metabolites, morphine and morphine-3-glucuronide (M-3-G), in rat plasma. Solid-phase extraction was used to separate codeine and its metabolites from plasma constituents. Extraction efficiencies of codeine, morphine and M-3-G from rat plasma samples were 97, 92 and 93%, respectively. The chromatographic separation was performed using a reversed-phase C18 column and an elution gradient at ambient temperature. Using native fluorescence detection (excitation at 245 nm and emission at 345 nm), the detection limits of 50 ng/ml for morphine, 25 ng/ml for codeine and 20 ng/ml for M-3-G were obtained. The method had good precision, accuracy and linearity, and was applied to the study of glutethimide's influence on codeine metabolism in rat, following single doses of codeine glutethimide association. The results confirmed the fact that glutethimide was responsible for a significant increase of morphine plasma levels and for their maintenance in time, concomitant with a significant decrease of M-3-G plasma levels, explained by the inhibition of morphine glucuronidation. In conclusion, glutethimide potentiates and prolongs the analgesic effect of codeine by a pharmacokinetic mechanism. PMID- 12899974 TI - Determination of ticlopidine in human plasma by capillary gas chromatography with ion trap detection. AB - A gas chromatographic method for determination of ticlopidine in human plasma using mass spectrometric detection was developed. A Perkin Elmer model 8500 gas chromatograph coupled to an ion trap detector (ITD) in electron impact mode (EI) was used. The gas chromatograph was fitted with a 30 m x 0.32 mm i.d., DB-17 capillary column with a film thickness of 0.25 microm. The compounds were isolated from plasma by Extrelut-1 solid-phase extraction using hexane as the solvent of elution. The detection and quantification limits for this method were 0.005 and 0.01 microg/ml, respectively. The calibration curves were linear from 0.01 and 2.5 microg/ml. Recoveries from human plasma spiked at 0.01 and 2.0 microg/ml ranged from 84.4 and 87.3%. The coefficient of variation (CV) was between 5.1 and 6.9%. The results show that the sensitivity, accuracy and precision of the method are adequate for the determination of ticlopidine in human plasma samples. The ITD in SIM mode allows the unequivocal identification of ticlopidine. The suitability of this method was evaluated in the determination of ticlopidine in plasma from healthy volunteers following oral administration of a dose of 500 mg/day. PMID- 12899975 TI - Simultaneous high-throughput determination of clenbuterol, ambroxol and bromhexine in pharmaceutical formulations by HPLC with potentiometric detection. AB - Potentiometric detection of clenbuterol, ambroxol and bromhexine in marketed pharmaceuticals was described in six isocratic HPLC systems. The podant- and macrocyclic-type neutral ionophores, N,N,N',N'-tetracyclohexyl-oxybis(o phenyleneoxy)diacetamide (TOPA) and hexakis(2,3,6-tri-O-octyl)-alpha-cyclodextrin (OCD), were applied in poly(vinyl)chloride (PVC)-based liquid membrane electrodes. Both types of neutral ionophores improve the sensitivity for all mentioned drugs when compared with a tetrakis(p-chlorophenyl)borate (BOR)-based electrode as well as with single wavelength UV detection. Detection limits (S/N=3) of 2.6 x 10(-10) mol l(-1) (injected concentration) for the highly hydrophobic bromhexine were achieved with the TOPA-based electrode and a cyano reversed-phase (RP)-HPLC with Uptisphere UP5CN-25QS silica column (250 x 4.6 mm i.d.) eluted with acetonitrile (AcN)-ethanol-perchloric acid (1.66 mM) (60:2:38, v/v/v) (pH* 2.45). Comparable result was obtained with OCD-based electrodes and an XTerra RP18 hybrid silica-polymer column eluted with AcN-phosphoric acid (20 mM) (25:75, v/v) (pH* 2.60). In the mobile phases containing 60-75% v/v AcN or methanol, stable and reproducible response of both types of neutral ionophore based electrodes was observed for at least 3 days. The results of the validated procedure for reliable simultaneous determination of the drugs in fortified representative samples of pharmaceuticals were also presented. PMID- 12899976 TI - Analysis of optically active compounds using conventional chromatography with a circular dichroism detector. AB - Analysis of optically active compounds in complex samples is often based on chiral chromatography or capillary electrophoresis in order to separate the enantiomers. This requires a chiral reagent, when using conventional chromatography, or an expensive chiral column, or a chiral selector, when using capillary electrophoresis. The type of column, reagent, or additive depends highly on the compound to be analysed. A simple and generally applicable method is using a conventional HPLC column coupled to a CD detector. Separation of enantiomers is not required, as they can be identified by a positive or negative peak. A racemate does not produce a peak; neither does an optically inactive compound. The application of HPLC-CD for the identification of pharmacologically active compounds, such as dexamphetamine, 5-hydroxytryptophan, (-)-huperzine A, and interferon, as standards, in registered drugs, in falsifications, and in food supplements is described. PMID- 12899977 TI - Comparison of morphine and hydromorphone analysis on reversed phase columns with different diameters. AB - A comparison of a reversed phase high-performance liquid chromatographic (HPLC) method performed on columns with different internal diameters is reported for the quantitative routine determination of morphine.HCl and hydromorphone.HCl in solutions used for intramuscular injection. The method is based on the ion pairing properties of sodium dodecyl sulphate (SDS) with alkaloids on a reversed phase LiChrospher RP-18 packing material and UV-detection at 230 nm. The mobile phase consisted of an acetonitrile: water mixture 35:65 (v/v) containing 0.5% (w/v) SDS and 0.4% (v/v) acetic acid. Precision, linearity and limit of detection were compared on the 2, 3 and 4 mm i.d. x 125 mm columns. A robustness test for the determination of hydromorphone.HCl was also evaluated. PMID- 12899978 TI - Simultaneous determination of methylparaben, propylparaben, hydrocortisone acetate and its degradation products in a topical cream by RP-HPLC. AB - A novel reversed-phase high-performance liquid chromatographic method with UV spectrophotometric detection was developed and validated for the determination of compounds in topical cream. The method describes determination of active component hydrocortisone acetate (HCA), its degradation products hydrocortisone (HC) and cortisone acetate (occurring in formulation after long-term stability tests) and two preservatives presented in the cream-methylparaben and propylparaben, using dexamethasone as an internal standard. The chromatographic separation was performed on a 5 microm SUPELCO Discovery C18 125 x 4-mm ID column. The optimised mobile phase for separation of all the compounds consists of methanol, acetonitrile and water (15:27:58, v/v/v), with the analysis time less than 13 min. The method was applicable for routine analysis (assays and stability tests) of active compound HCA, preservatives and degradation products in pharmaceutical product--topical cream Hydrocortizone cream 1%. PMID- 12899979 TI - Photochemical degradation of solid-state nisoldipine monitored by HPLC. AB - The photochemical degradation of solid-state nisoldipine, 1,4-dihydropyridine calcium antagonist, was investigated under daylight and UV light conditions. Degradation products were identified by using the retention times of corresponding standards and quantified by high-performance liquid chromatographic method. The daylight illumination induced appearance of nitrosophenylpyridine, while formation of second degradation product, nitrophenylpyridine, was observed only upon UV light illumination. The photodegradation kinetics of solid-state nisoldipine under daylight and UV light illumination belongs to class of zero order reactions. The rate constants of disappearance of nisoldipine upon illumination were determined for raw material as well as pharmaceuticals (tablets, film-tablets and capsules). PMID- 12899980 TI - Capillary electrophoresis in analysis of veterinary drugs. AB - Antibiotics are extensively applied in veterinary medicine for the treatment of various bacterial infections. Because of their use in food producing animals, the risk of occurrence of unwanted residues in edible products exists. To ensure human food safety, The European Union has defined maximum residue limits (MRLs) for veterinary drug residues in food products. Analytical methods need to be developed to confirm the presence of antibiotics at the MRL level. A capillary electrophoresis (CE) method with UV detection is proposed for the quantitative determination of residues from poultry and porcine tissues. Eight of the most frequently used antibiotics and nifursol, which routinely used as poultry coccidiostat, were analysed. CE technique permitted analysing substances to be separated from muscle, liver, kidney and skin with fat after a simple extraction with acetonitrile or ethyl acetate under basic conditions. Proposed method is capable to identify drug residues in tissues at level below 20 microg/kg. PMID- 12899981 TI - Determination of ursodeoxycholic acid in pharmaceutical preparations by capillary electrophoresis with indirect UV detection. AB - A simple and rapid capillary electrophoretic method, with indirect UV detection, for the quantification of ursodeoxycholic acid (UDCA) in pharmaceutical preparations was developed in this study. Sodium p-hydroxy benzoate was used as background electrolyte (BGE) (5 mM, pH 8.0) and visualization agent. Separation was carried out on a fused-silica capillary (50 microm x 72 cm) at a potential of 25 kV under ambient temperature and detected at 250 nm. Glycocholic acid was used as internal standard for quantification. Both run-to-run repeatability and day-to day reproducibility of migration time were below 0.1% relative standard deviation (R.S.D.). Repeatability and reproducibility of relative peak height were 3.3 and 3.8% R.S.D., respectively. Accuracy was tested by spiking two pharmaceutical tablets with standards and the recoveries were 101.9+/-9.87 and 99.6+/-9.60% (n=3), respectively. Linearity of relative peak height was tested in the range 20 100 microg/ml. Limit of detection (LOD) was 3 microg/ml. The method could be used to assay UDCA raw materials and formulation products. PMID- 12899982 TI - Adsorptive stripping voltammetric determination of cefetamet in human urine. AB - On the basis of previously established mechanism of cefetamet (CEF) reduction, two methods were suggested for CEF determination-differential pulse polarographic and differential pulse adsorptive stripping voltammetric method. Two pH values were chosen, 2.0 and 8.4, where the electrochemical process was defined as one four-electron and two two-electron processes, respectively. The methods were performed in Britton-Robinson (BR) buffer and the corresponding calibration graphs were constructed and statistical parameters were evaluated. Applying the AdSV method at pH 2.0 linearity was achieved from 2 x 10(-8) to 2 x 10(-7) M with limit detection and limit determination of 4 x 10(-9) and 1.4 x 10(-8) M, respectively. At pH 8.4, the linearity was obtained between 6 x 10(-8) and 6 x 10(-7) M, with limit detection and limit determination of 1.5 x 10(-8) and 5 x 10(-8) M, respectively. Since the AdSV method enabled lower concentrations of CEF to be determined, this method was tested for CEF determination in spiked urine samples, and DPP method was used as a comparative one. PMID- 12899983 TI - Voltammetric and electrochemical quartz crystal microbalance studies of sulforaphane and its Zn(II) complexes. AB - Cyclic voltammetry (CV) and electrochemical quartz crystal microbalance (EQCM) methods were used to study the behavior of sulforaphane (SFR). To established electrochemical properties of SFR methyl-, ethyl-, 2-chloroethyl-, propyl-, butyl and tert-butyl-isothiocyanates were examined at gold electrode. Optimal measurement parameters were established and stock solutions developed. The electrode activity of the studied compounds (cathodic peak E(p)=-0.6 V) was found to be primarily due to the oxidation of the isothiocyanate group. By the EQCM method it was found that during the reduction of SFR the mass of the electrode systematically increased in successive scans by ca. 120 ng per scan and in the mixture of SFR and Zn(2+) ions ca. 350 ng. Formation of SFR complexes with Zn(2+) ions was confirmed by EQCM and UV methods. Cyclic voltammetric method has been elaborated for SFR determination and compared with UV method. PMID- 12899984 TI - Development of electrochemical methods for determination of tramadol--analytical application to pharmaceutical dosage forms. AB - A square-wave voltammetric (SWV) method and a flow injection analysis system with amperometric detection were developed for the determination of tramadol hydrochloride. The SWV method enables the determination of tramadol over the concentration range of 15-75 microM with a detection limit of 2.2 microM. Tramadol could be determined in concentrations between 9 and 50 microM at a sampling rate of 90 h(-1), with a detection limit of 1.7 microM using the flow injection system. The electrochemical methods developed were successfully applied to the determination of tramadol in pharmaceutical dosage forms, without any pre treatment of the samples. Recovery trials were performed to assess the accuracy of the results; the values were between 97 and 102% for both methods. PMID- 12899985 TI - Analytical methods for the determination of folic acid in a polymeric micellar carrier. AB - Amphiphilic copolymers have been the object of growing scientific interest due to their ability to form polymeric micelles in aqueous environments entrapping lipophilic drugs in their inner core. In this study, polyvinylalcohol substituted with oleic acid was employed as an amphiphilic micellar carrier for folic acid (FA), a model drug similar for its chemical-physical characteristics to methotrexate. In order to investigate the stability of the polymeric micelles, the drug incorporation and the kinetic aspects of drug release from these systems, selective analytical methods are required. The development of three analytical methods suitable for selectively identifying and reliably determining FA contained in the micelles and in the delivery systems is reported. UV derivative (first and second order) spectrophotometry was first applied to the aqueous solution of the FA containing micelles obtained at pH 9.0 and provided a characteristic spectral profiling with sharp peaks, related to the analyte, whose amplitude was used for quantitative application. A second approach involved a solid phase extraction (strong anion exchanger), which provided an effective clean up of the FA micelles solution, allowing accurate analysis to be performed also by a conventional spectrophotometric method. A RP-HPLC method, selectively supplying the FA separation from the micelles' components, was then used as a reference method to determine the accuracy of the spectrophotometric methods. These methods were applied to various micelle composition and to the delivery system study. PMID- 12899986 TI - Detection and characterization of cyclodextrin complexes with beta-carboline derivatives by spectroscopic techniques. AB - beta-Carboline alkaloids exhibit a great variety of pharmacological activities. The solid inclusion complexes of harmane and harmine with beta-cyclodextrin and also with hydroxypropyl-beta-cyclodextrin, have been prepared following different procedures. IR and NMR spectroscopies were employed to verify the interaction of the guest molecules with the cyclodextrin cavities. The differences observed in the IR and NMR spectra are in agreement with those described in the literature for other guest molecules. The shifts in the 13C- and 1H-NMR spectra confirm the existence of the inclusion complexes. The fluorescence emission spectra of these complexes dissolved in buffered aqueous solution (pH 7.3) exhibit the characteristic peaks of the cationic form for harmane alkaloids. The neutral bands are not present for the free alkaloids in aqueous solutions. Fluorescence quenching emission of the complexes is compared to that of the corresponding free alkaloids. PMID- 12899987 TI - Spectrophotometric determination of cimetidine in pharmaceuticals and urine using batch and flow-injection methods. AB - Two sensitive and fast spectrophotometric methods using batch and flow-injection procedures for the determination of cimetidine (CMT) are proposed. The methods are based on the formation of a green complex between this drug and Cu(II) in acetic/acetate medium of pH 5.9. The calibration graphs resulting from measuring the absorbance at 330 nm are linear over the ranges 2.5 x 10(-6)-1.0 x 10(-3) and 5 x 10(-6)-2.0 x 10(-3) M with detection limits of 9.5 x 10(-7) and 2.1 x 10(-6) for batch and flow-injection methods, respectively. The methods are applied to the routine analysis of CMT in pharmaceuticals and human urine. PMID- 12899988 TI - Multi-pumping flow system for the spectrophotometric determination of dipyrone in pharmaceutical preparations. AB - A novel flow system for the spectrophotometric determination of dipyrone with p dimethylaminobenzaldehyde exploiting the multi-pumping approach was developed. The proposed methodology utilises several micro-pumps for propelling the involved fluids under improved mixing conditions, introducing sample/reagent aliquots and providing commuting facilities. As a consequence the multi-pumping system presents high versatility and manifold simplicity, as well as a straightforward operational control and enhanced analytical capabilities. Linearity of the analytical curve was observed within 10 and 400 mg l(-1) dipyrone (r=0.9997; n=6), results were precise (r.s.d.<0.12%; n=20) and sampling rate was 50 h(-1). Detection limit was estimated as 1 mg l(-1) dipyrone. The method was applied to pharmaceutical preparations and the results were in agreement with those obtained by the reference procedure with relative deviations within -1.7 and +2.2%. PMID- 12899989 TI - First-order UV-derivative spectrophotometry in the analysis of omeprazole and pantoprazole sodium salt and corresponding impurities. AB - The first-order UV-derivative spectrophotometry, applying zero-crossing method was developed for the determination of omeprazole (OM), omeprazole sulphone (OMS), pantoprazole sodium salt (PANa), and N-methylpantoprazole (NPA) in methanol-ammonia 4.0% v/v, where the sufficient spectra resolutions of drug and corresponding impurity were obtained, using the amplitudes 1D(304), 1D(307), 1D(291.5) and 1D(296.5), respectively. Method showed good linearity in the ranges (microg ml(-1)): 1.61-17.2 for OM; 2.15-21.50 for OMS; 2.13-21.30 for PANa and 2.0-20.0 for NPA, accuracy and precision (repeatability and reproducibility). The experimentally determined values of LOD (microg ml(-1)) were 1.126; 0.76; 0.691 and 0.716 for OM, OMS, PANa and NPA, respectively. The obtained values of 2.91% w/w for OMS and 3.58% w/w for NPA in the presence of their parent drug, by applying the method of standard additions, point out the usage of the proposed method in stability studies. Zero-crossing method in the first-order derivative spectrophotometry showed the impurity-drug intermolecular interactions, due to the possible intermolecular hydrogen bonds, confirmed by divergences of experimentally obtained amplitudes for impurities OMS and NPA in comparison to expected values according to regression equations of calibration graphs. PMID- 12899990 TI - Specific spectrophotometric method with trifluoroacetic acid for the determination of selenium(IV) in selenitetriglycerides. AB - The role of selenium as an antioxidant and anticancer agent is very well documented in the literature. Selenium compound showing the highest activity as a free radicals scavenger and as an anticancer agent should contain selenium at +4 oxidation level. The synthesis of selenitetriglycerides (named selol) was carried out in the Department of Drug Analysis at Warsaw Medical University (Polish Patent 1999). Selenitetriglycerides showed a dimeric structure. In a single dose toxicity studies performed in rats, LD50 was 100 mg Se kg(-1) after oral administration of selol. The subcutaneous and intraperitoneal administration of selol showed extremely low toxicity. The aim of this work was to develop a new specific method for the determination of Se(IV) in selol. We stated that selenitetriglycerides react quantitatively with trifluoroacetic acid (TFA) in dichloromethane giving a red-coloured conjugate. However, recorded spectrum showed the maximum absorption in the wavelength 380 nm. The optimal conditions of the reaction were established, namely temperature 35 degrees C and reaction time 35 min. The reaction was proved to be specific because neither selenites nor other selol constituents react with TFA. The constructed calibration curve obeyed the Lambert-Beer law in the range of 0.1-7.4 mg ml(-1). Molar absorption coefficient is epsilon =9.46 x 10(3) l mol(-1) cm(-1) and epsilon =2.36 x 10(5) l mol(-1) cm(-1) calculated for selenium and selenitetriglyceride dimer (m.w. 1972.72), respectively. Obtained results for selenium determination were confirmed by AAS method. The developed method showed specificity and high sensitivity. PMID- 12899991 TI - Quality control of commercial tablets containing the novel antipsychotic quetiapine. AB - Quetiapine (bis [2-(2-[4-(dibenzo[b,f][1,4]thiazepin-11 yl]ethoxy)ethanol]fumarate) is the most recent agent introduced on the drug market for the treatment of psychotic disorders. Two different analytical methods for the quality control of quetiapine in commercial formulations have been developed and compared: a spectrophotometric method and a capillary zone electrophoretic (CZE) method. The spectrophotometric assay was carried out measuring the absorbance at a wavelength of 246 nm. The CZE method used an uncoated fused-silica capillary and a pH 2.5, 50 mM phosphate buffer as the background electrolyte. The detection wavelength was 205 nm, the separation voltage was 15 kV, and a complete electrophoretic run lasts less than 2.5 min. Extraction of quetiapine from the commercial tablets consisted of a simple one step treatment with a pH 2.5, 50 mM phosphate buffer. Linearity was observed in the 5-25 microg ml(-1) concentration range of quetiapine for the spectrophotometric method, and in the 5-50 microg ml(-1) concentration range for the electrophoretic method. Both methods gave satisfactory results in terms of repeatability and intermediate precision (RSD<1.9%). Also accuracy values were very good for both methods, the recovery being between 98.2 and 100.5%. PMID- 12899993 TI - Enantiomeric resolution of some 2-arylpropionic acids using L-(-)-serine impregnated silica as stationary phase by thin layer chromatography. AB - The enantiomeric resolution of certain 2-arylpropionic acids was achieved on thin silica gel plates impregnated with optically pure L-(-)-serine as chiral selector. The mobile phase enabling successful resolution of (+/-)-ibuproxam and (+/-)-ketoprofen was acetonitrile-methanol-water (16:4:0.5, v/v/v) and (16:3:0.5, v/v/v) for (+/-)-tiaprofenic acid. The spots were detected with iodine vapors and the detection limits were found to be different for each of the 2-arylpropionic acid, ranging between 0.25 and 0.50 microg/ml. The effect of concentration of the impregnating chiral selector, temperature and pH on resolution has been studied. The procedure was applied successfully to resolve commercial ampoules of ketoprofen dosage formulation. PMID- 12899992 TI - Herbal remedies of Solidago--correlation of phytochemical characteristics and antioxidative properties. AB - In this study the correlation of phytochemical characteristics and antioxidative properties of classical herbal tea extracts-Infusum solidaginis, Decoctum solidaginis, Maceratum solidaginis-and tinctures prepared by various concentration of ethanol (40, 70, 96% v/v) have been examined for the release of flavonoids and their antioxidant activity. Quantitative and composition determination of flavonoids were carried out by spectrophotometry, high performance liquid chromatography and capillary electrophoresis, respectively. Hydrogen-donating ability and reducing power properties were used to define in vitro radical scavenging activity of Solidago extracts, but integral antioxidative capacity was determined by luminometry (Photochem), calculating the ascorbic acid equivalents. Chlorogenic acid, quercetin-3-O-beta-D-rutinoside, quercetin-3-O-beta-D-galactoside, quercetin-3-O-beta-D-glucoside, quercetin-3-O beta-D-rhamnoside, kaempferol-3-O-alpha-L-rhamnoside and quercetin were confirmed by retention times and UV spectra. Based on the dissolution rate, variance of flavonoid release and ascorbic acid equivalents it was concluded, that Tinctura solidaginis (70% v/v ethanol) and Infusum solidaginis are the most appropriate preparations. PMID- 12899994 TI - HPLC determination of tramadol in human breast milk. AB - Tramadol is a centrally acting analgesic used for prevention and treatment of moderate to severe pain. It is estimated that 0.1% of the administered dose passes into breast milk causing potentially unwanted effects in nursing babies. Pharmacokinetically, breast milk is supposed to be a separate compartment into which the drug is excreted-mainly by passive diffusion. Due to a complex composition of breast milk, a suitable sample preparation procedure is needed with a subsequent chromatographic analysis for drug determination. Among several sample cleanup procedures tested we chose the liquid-liquid extraction procedure using n-hexane as an organic phase with back extraction into aqueous phase since it was considered the most suitable and the most compatible with the subsequent HPLC analysis. The precision and the reproducibility of the method were improved approximately two times by using metoprolol as an internal standard thus making the method also more robust with regard to a variable composition of milk samples. These characteristics, together with low detection limit and short analysis time, proved that the developed method is suitable for monitoring of tramadol in human breast milk. PMID- 12899995 TI - Degradation of components in drug formulations: a comparison between HPLC and DSC methods. AB - Information about the stability of drug components and drug formulations is needed to predict the shelf-life of the final products. The studies on the interaction between the drug and the excipients may be carried out by means of accelerated stability tests followed by analytical determination of the active principle (HPLC and other methods) and by means of the differential scanning calorimetry (DSC). This research has been focused to the acetyl salicylic acid (ASA) physical-chemical characterisation by using DSC method in order to evaluate its compatibility with some of the most used excipients. It was possible to show, with the DSC method, the incompatibility of magnesium stearate with ASA; the HPLC data confirm the reduction of ASA concentration in the presence of magnesium stearate. With the other excipients the characteristic endotherms of the drug were always present and no or little degradation was observed with the accelerated stability tests. Therefore, the results with the DSC method are comparable and in good agreement with the results obtained with other methods. PMID- 12899996 TI - Fast drug stability determination by LC variable-parameter kinetic experiments. AB - Variable-parameter kinetic experiments were carried out using HPLC as analytical instrument. The hydrolysis of aspirin was followed both at variable-temperature and at variable-pH conditions. The peak areas relative to salicylic acid were processed by direct fit to a mathematical model and/or by differential method obtaining, by single experiments, the values of the apparent rate constant in the whole range of temperature and pH studied. The results, although the discontinuity of this kind of analysis, are in agreement with those obtained by constant-parameter kinetics but saving experimental time. PMID- 12899997 TI - Determination of 4-aminophenol impurities in multicomponent analgesic preparations by HPLC with amperometric detection. AB - A method for the determination of 4-aminophenol, the main impurity of paracetamol, by high-performance liquid chromatographic (HPLC) method with amperometric detection has been developed. The analysis was performed in an isocratic mode on a reversed phase Luna column 5 microm C-18 (100 x 4.6 mm). A mobile phase (0.05 mol l(-1) LiCl solution containing 18% methanol adjusted to pH 4.0 with orthophosphoric acid) was suitable for the separation and determination of 4-APh. Chromatograms were recorded for 250 s by means of an amperometric detector at a potential of +325 mV of the glassy carbon electrode versus the reference electrode Ag/AgCl. The proposed liquid chromatographic method was successfully applied to the analysis of commercially available multicomponent dosage forms. The sensitivity of the detection for 4-aminophenol was 1 ng ml(-1) for substance and 4 ng ml(-1) for tablets or capsules. The method developed in this study is sensitive and selective and can be applied for routine studies of pharmaceuticals in the form of tablets or capsules. PMID- 12899998 TI - Simultaneous determination of naproxen and related compounds by HPLC using porous graphitic carbon column. AB - A simple, selective and sensitive high performance liquid chromatographic (HPLC) method has been developed for the simultaneous determination of naproxen and its main degradation products such as 1-(6-methoxy-2-naphthyl) ethanol (MNE), 2 methoxy-6-ethyl naphthalene (MEN) and 2-acetyl-6-methoxy naphthalene (AMN). The separation of these compounds was achieved on porous graphitic carbon (PGC) column using tetrahydrofuran-methanol as the mobile phase, and the effluent from the column was monitored at 272 nm. At a flow rate of 1 ml min(-1), the retention time of the last eluting compound was less than 10 min. Correlation coefficient for calibration curves in the ranges 2-25 microg ml(-1) for all compounds studied were greater than 0.999. The sensitivity of detection is 0.05 microg l(-1) for naproxen, MNE and MEN and 0.20 microg ml(-1) for AMN. The reproducibility of the peak area of these compounds using isocratic elution were quite high, and the standard deviations (S.D.) were below 2% (n=5). The reproducibility of retention times of these compounds was within 1% (n=5). The proposed liquid chromatographic method was successfully applied to the analysis of commercially available naproxen sodium (NS) dosage forms with recoveries of 98.8-102%. A comparative study shows that the selectivity of these compounds on PGC column was different to that obtained with octadecyl silica (ODS) columns. PMID- 12899999 TI - A new proposal for fast determination of vitamin B2 from aqueous pharmaceutical products. PMID- 12900001 TI - Drug analysis 2002. Proceedings of a joint meeting of the 7th International Symposium on Drug Analysis and the 13th International Symposium on Pharmaceutical and Biomedical Analysis. April 21-25, 2002. Bruges, Belgium. PMID- 12900000 TI - Serum levels of beta-trace protein and glomerular filtration rate--preliminary results. AB - The aim of this study was to evaluate the relationship between serum levels of beta-trace protein (BTP), a low molecular weight (MW) protein, and glomerular filtration rate (GFR). GFR and serum levels of BTP, and for comparison creatinine (Creat), cystatin C (Cys) and beta 2-microglobulin (beta 2M), were measured in 60 patients, with renal function ranging from normality to advanced renal failure. Serum levels of BTP progressively increased with the reduction of GFR. A good correlation was found between GFR and serum levels of BTP (r=0.918), Creat (r=0.932), Cys (r=0.937), and beta 2M (r=0.924). Furthermore, no statistically significant difference was found between BTP and Creat, Cys, beta 2M, as indicators of a moderate GFR impairment. These preliminary data indicate that BTP might be suitable as an indicator of GFR. PMID- 12900003 TI - The new World Health Organization/International Society of Urological Pathology (WHO/ISUP) classification for TA, T1 bladder tumors: is it an improvement? AB - The World Health Organization/International Society of Urological Pathology (WHO/ISUP) consensus classification of urothelial neoplasms of the urinary bladder was developed in an attempt to both improve upon prior classification systems as well as to adopt a classification system that would have widespread acceptance. Prior to this classification system, numerous diverse grading schemes for bladder cancer existed whereby the same lesion seen by different pathologists would result in very different diagnoses solely based on definitional differences of lesions. Another strength of the consensus classification system is that it provides detailed histological criteria for papillary urothelial lesions. In contrast, prior grading systems for bladder tumors were vague and subjective. The current classification system allows for designation of a lesion (papillary urothelial neoplasm of low malignant potential), which biologically has a very low risk of progression, yet is not entirely benign. In the past, these lesions were a source of controversy, as some experts in the field required very restrictive criteria for the diagnosis of papilloma and would label such lesions as malignant. Other experts in the field, not wanting to label a patient with such a low-grade papillary lesion as having carcinoma, would diagnose these lesions as papilloma. This intermediate category allows both schools of thought to diagnose a lesion as not fully malignant, yet still documents need for additional follow-up. Since its inception, several studies have been published documenting the relationship of tumors classified using the WHO/ISUP system to prognosis. These articles are summarized within this review. PMID- 12900004 TI - Current bladder cancer tests: unnecessary or beneficial? AB - Bladder cancer is currently diagnosed using cystoscopy and cytology in patients with suspicious signs and symptoms. These same tests are used to monitor patients with a history of bladder cancer for recurrence. The recurrence rate for bladder cancer is high, thus necessitating long-term follow-up. Urine cytology requires an experienced cytopathologist and is costly. It has high specificity, but low sensitivity for low-grade bladder tumors. Recently many non-invasive bladder cancer tests, utilizing markers found in the urine, have been developed. The FDA has approved several of these for the use is bladder cancer diagnosis, and many others are undergoing development and investigation. An ideal bladder cancer test would be non-invasive, highly sensitive and specific, inexpensive, easy to perform, and yield highly reproducible results. Many of the tests reviewed meet some, but not all, of these criteria. PMID- 12900005 TI - Intravesical therapy of superficial bladder cancer. AB - The aim of treatment of superficial bladder cancer with intravesical therapy is threefold: (1) Eradicate existing disease. (2) Prevention of recurrence. (3) Prevention of tumor progression. The prognostic factors allow differentiation in different risk groups and this is useful in planning treatment. Studies on pharmacokinetics have proved the efficacy of optimized drug delivery. Comparing resection with and without intravesical chemotherapy a short term approximately 15% decrease in tumor recurrence with chemotherapy can be obtained but no effect on progression was proven. No agent has proved more effective than the other. Single, early instillation of chemotherapy has proven effective but the role of maintenance therapy has been controversial. Immunotherapy in the form of Bacillus Calmette-Guerin generally have proven more efficacious than chemotherapy. The results in comparison to mitomycin C have not been as conclusive. Several new approaches are explored to improve the efficacy of this therapy. PMID- 12900006 TI - Combination of intravesical chemotherapy and hyperthermia for the treatment of superficial bladder cancer: preliminary clinical experience. AB - The prevalence of superficial transitional cell carcinoma of the bladder (STCCB) is still increasing in spite of improved adjuvant chemotherapeutic and/or immunoprophylaxis approaches. Thus, there is certainly an urgent need to improve our ability to control this disease. Local hyperthermia has a therapeutical potential for the treatment of many solid tumors, especially when used in combination with other treatments, such as radiation and chemotherapy. In particular, a synergistic or, at least, supra-additive anti-tumor cell killing effect was documented when local hyperthermia was administered in combination with selected cytostatic drugs. Recently, advances in miniaturized technology have allowed the development of a system specifically designed for delivering an endovesical thermo-chemotherapy regimen in humans. In preliminary clinical experiences, insofar mainly carried out as mono-institutional investigations, the combined treatment using this system was demonstrated to be feasible, minimally invasive and safe when performed on out-patient basis. Moreover, the anti-tumoral efficacy seemed to be significantly enhanced when compared with that obtained using intravesical chemotherapy alone for both adjuvant (prophylaxis) and neo adjuvant (ablative) approaches to superficial bladder cancer. PMID- 12900007 TI - Radical cystectomy with or without urethrectomy? AB - After the introduction of bladder replacement procedures, the indications for prophylactic urethrectomy have become more and more restricted. While years ago, a prophylactic urethrectomy was performed in many patients with cutaneous diversions, it has become clear that only patients with invasion by transitional cell carcinoma at the level of the prostatic urethra or bladder neck have a substantial risk of developing subsequent urethral recurrence. The pre- or intra operative assessment of the prostatic urethra in males and of the bladder neck in females is the key to appropriate management of the urethra in patients with bladder cancer. PMID- 12900008 TI - Orthotopic bladder reconstruction in women--what we have learned over the last decade. AB - Approximately 10 years ago protocols for urethra-sparing cystectomy and orthotopic urinary diversion to the urethra in female patients with bladder cancer were initiated at several centers. Long-term data regarding the oncological and functional outcome are the subject of this review. Studies regarding the relationship between primary bladder cancer and secondary urethral tumors in women revealed in most studies a lower risk for women than for men in most studies. In a recent meta-analysis the incidence of urethral tumors was 6.8% in 5657 male and 3.6% in 841 female patients with transitional cell cancer of the bladder. Anatomical and functional studies revealed that smooth musculature can be found in the entire length of the female urethra. The rhabdosphincter which is the important structure for postoperative continence in low pressure intestinal reservoirs is in the midportion of the urethra which will not be touched during urethra-sparing surgery. A recent study looked at the oncological and functional results of 102 women with orthotopic urinary diversion after a follow-up ranging from one and half to 100 months (mean 26, median 24 months). There was no perioperative mortality, and an early and late complication rate requiring secondary intervention in 5 (5%) and 12 (12%) patients. With 88 of 102 patients alive and 83 of 102 patients disease free, a disease specific survival of 74% and a disease free survival of 63% was estimated at 5 years. No pelvic recurrence was seen in 81 patients with TCC. Daytime continence was 82%; nocturnal continence was 72%. Twelve patients (12%) were unable to empty their bladders completely and needed some form of catheterization. Increasing experience in recent years confirms the initial preliminary results showing that sparing the urethra at cystectomy will not compromise oncological outcome and can be satisfactorily used for orthotopic reconstruction of the lower urinary tract. Both diurnal and nocturnal continence and clean intermittent catheterization rates after 6 months justify the use of orthotopic neobladders as the procedure of choice in the majority of female patients with bladder neoplasms. PMID- 12900009 TI - Transitional cell carcinoma of the ureter and renal pelvis. AB - Transitional cell carcinoma (TCC) of ureter and renal pelvis is relatively uncommon. Smoking, occupational carcinogens, analgesic abuse, Balkan nephropathy are the risk factors. Cytogenetic studies revealed that the most frequent aberration is the partial or complete loss of chromosome 9. Approximately 20-50% of patients with upper urinary tract (UUT) TCC have bladder cancer at some point on their course, whereas the incidence of UUT TCC after primary bladder cancer is 0.7-4%. Excretory urography and retrograde pyelography are the conventional diagnostic tools; however, ureteropyeloscopy combined with cytology and biopsy is more accurate. Grade and stage of the disease have the most significant impact on survival. Nephroureterectomy with bladder cuff excision has been the mainstay of treatment. Local resection may be appropriate for distal ureteral lesions especially when the disease is low grade and stage. Advances in endourology have made it possible to treat many tumors conservatively. Ureteroscopic and to a certain extent percutaneous surgical approaches are widely used today especially in patients with low grade, low stage disease. Endoscopic close surveillance is mandatory for these patients. Adjuvant topical therapies appear to be safe but confirmation of any benefits awaits the results of further large studies. More recently, laparoscopic techniques have become a viable alternative to open surgery, but long term cancer control data are lacking. Aggressive surgical resection does not affect the outcome of patients with advanced disease. Adjuvant radiotherapy is ineffective, and systemic chemotherapy results in a low complete response rate for patients with metastases. PMID- 12900010 TI - Is there standard chemotherapy for metastatic bladder cancer? Quality of life and medical resources utilization based on largest to date randomized trial. AB - A large multinational phase III trial performed during the late 1990s compared two systemic chemotherapy regimens gemcitabine plus cisplatin and methotrexate, vinblastine, doxorubicin, cisplatin (M-VAC) in more than 400 patients with advanced or metastatic urothelial cancer. This trial has been discussed to landmark the beginning of a new era following M-VAC polychemotherapy which has dominated treatment of advanced urothelial cancer throughout the previous decade. Despite the fact that gemcitabine/cisplatin combination therapy did not surpass M VAC therapy in regard to patient survival as initially intended, this combination demonstrated a more favourable toxicity profile with improved tolerability and superior cost effectiveness, rendering this combination an attractive alternative to M-VAC. This review on the largest to date phase III trial for advanced or metastatic urothelial cancer will focus on issues of quality of life including indicators such as performance status and changes in body weight. Furthermore, data on medical resources utilization as accounted during systemic polychemotherapy and related toxic events will be reconsidered, particularly under the impression of decreasing health care resources worldwide. PMID- 12900011 TI - Molecular biology of transitional cell carcinoma. AB - Transitional cell carcinoma (TCC) is the fifth most common solid malignancy in the USA. Radical cystectomy will cure a substantial fraction of patients with minimally invasive TCC, but approximately 50% of patients with muscle-invasive or extravesical disease treated by radical cystectomy alone die of metastatic TCC. Transitional cell carcinoma have a diverse collection of biologic and functional characteristics. This is reflected in a differing clinical course. The limited value of established prognosticators, make the analysis of new molecular parameters of interest in predicting the prognosis of bladder cancer patients; in particular, high-risk patient groups that are at risk of progression and recurrence. Over the past decade, there had been major progress in the elucidation of the molecular genetic changes leading to the development of TCC. This review will highlight these important advances and emphasize the ways in which molecular biology is likely to affect the development of future therapies. PMID- 12900012 TI - New drugs and new approaches in metastatic bladder cancer. AB - The median survival of patients with metastatic cancer of the urothelium who receive best supportive care only in 4-6 months. With the introduction of combination chemotherapy regimens including cisplatin and methotrexate for the management of metastatic urothelial cancer, median overall survival has doubled. Nevertheless, death due to cancer ultimately occurs in more than 80% of these patients, thus more effective therapy is required. The new available treatment modalities range from new combinations of conventional chemotherapeutic agents to combinations incorporating novel drugs like gemcitabine and the taxanes. These new combinations incorporate the new active agents in two, three or multiple drug combinations, administered either in one regimen or sequentially in various combinations and schedules intended to improve the outcome of bladder cancer patients. Ongoing phase III studies will help to define the role of these new combinations in the treatment of advanced bladder cancer. The improved understanding of the molecular biology of urothelial malignancies is helping to define the role of new prognostic indices that can direct the most appropriate choice of treatment for advanced disease. In addition, advances in the molecular biology of urothelial malignancies may allow identification of specific genetic lesions and biochemical pathways upon which future therapeutic approaches can be focused. The integration of newer biologic agents, probably to supplement rather than to supplant chemotherapeutic drugs, should be a primary direction of research with the objective to interfere with multiple aspects of bladder cancer progression. PMID- 12900014 TI - Regulation of capsule biosynthesis in serotype A strains of Pasteurella multocida. AB - The capsule of Pasteurella multocida serotype A strain ATCC 11039 is composed of hyaluronic acid and is an important virulence factor. Repeated subculturing of certain capsular serotype A strains results in dissociation from a capsulated to a noncapsulated phenotype with a concomitant loss of virulence. Although noncapsulated variants have been thought to arise as a result of mutation, the molecular mechanisms underlying this event are unknown. In this study, we demonstrate that restoration of the capsulated phenotype occurs in vivo subsequent to intraperitoneal inoculation of BALB/c mice with a noncapsulated variant. Moreover, reverse transcription polymerase chain reaction analysis revealed the capsule locus to be under transcriptional control. Cloning and sequencing of a 290-bp fragment within the promoter containing intergenic region of the capsule locus of 11039/iso revealed no significant alterations occurred subsequent to subculturing. These results demonstrate that serotype A P. multocida strain ATCC 11039 regulates capsule expression in response to an unidentified environmental factor(s), thereby providing insights into the molecular mechanisms underlying colonial dissociation. PMID- 12900013 TI - New members of the Escherichia coli sigmaE regulon identified by a two-plasmid system. AB - A previously established method, based on a two-plasmid system, was used to identify promoters recognized by RNA polymerase containing the extracytoplasmic stress response sigma factor sigmaE in Escherichia coli. In addition to previously identified rpoE-dependent promoters, 11 new promoters potentially directing the expression of 15 genes were identified that were active only after over-expression of rpoE. The promoters were confirmed and transcriptional start points of the promoters were determined by primer extension analysis and S1 nuclease mapping. All the promoters contained sequences similar to the consensus sequence of rpoE-dependent promoters. The new rpoE-dependent promoters governed expression of genes encoding proteins involved in primary metabolism (fusA, tufA, recR), phospholipid and lipopolysaccharide biosynthesis (psd, lpxP), signal transduction (sixA), proposed inner or outer membrane proteins (bacA, sbmA, smpA, yeaY), and proteins with unknown function (ybaB, yaiW, yiiS, yiiT, yfeY). PMID- 12900015 TI - Identification of a transmissible plasmid from an Argentine Sinorhizobium meliloti strain which can be mobilised by conjugative helper functions of the European strain S. meliloti GR4. AB - We describe in this work the identification and the conjugal properties of two cryptic plasmids present in the strain Sinorhizobium meliloti LPU88 isolated from an Argentine soil. One of the plasmids, pSmeLPU88b (22 kb), could be mobilised from different S. meliloti strains to other bacteria by conjugation only if the other plasmid, pSmeLPU88a (139 kb), was present. This latter plasmid, however, could not be transferred via conjugation (frequency <10(-9) transconjugants per recipient) contrasting with the conjugal system from the previously described strain GR4, where one plasmid is mobilisable and a second one (helper) is self transmissible. Despite the differences between the two systems, the conjugative helper functions present in the cryptic plasmids of strain GR4 were active in the mobilisation of plasmid pSmeLPU88b from strain LPU88. Contrasting with this, plasmid pSmeLPU88b was not mobilised by the helper functions of the broad-host range plasmid RP4. Eckhardt gel analysis showed that none of the plasmids from strain GR4 were excluded in the presence of plasmid pSmeLPU88b suggesting that they all belong to different incompatibility groups for replication. The small plasmid from strain LPU88, pSmeLPU88b, was only able to replicate in members of the Rhizobiaceae family such as Rhizobium leguminosarum, Rhizobium tropici and Agrobacterium tumefaciens, but not in Escherichia coli or Pseudomonas fluorescens. The observation suggests that most likely plasmid pSmeLPU88b was not received from a phylogenetically distant bacterium. PMID- 12900016 TI - Contribution of intracellular calcium stores to an increase in cytosolic calcium concentration induced by Mannheimia haemolytica leukotoxin. AB - The contribution of intracellular calcium stores to Mannheimia haemolytica leukotoxin (LKT)-induced increase in cytosolic calcium concentration was studied by pharmacologically inhibiting transport of calcium across the plasma and endoplasmic reticulum membranes of bovine neutrophils exposed to LKT. Active intracellular storage of calcium by sarcoplasmic/endoplasmic reticulum calcium ATPase, influx of extracellular calcium across the plasma membrane, and release of stored calcium via inositol triphosphate receptors and ryanodine-sensitive calcium channels were inhibited using thapsigargin, lanthanum chloride, xestospongin C, and magnesium chloride, respectively. Pre-incubation with thapsigargin attenuated the increase in cytosolic calcium concentration produced by LKT, thus confirming the involvement of intracellular calcium stores. Inhibitory effects of lanthanum chloride, xestospongin C, and magnesium chloride indicated that the increase in cytosolic calcium concentration induced by LKT resulted from both influx of calcium across the plasma membrane and release of calcium from intracellular stores. PMID- 12900017 TI - Utilization for lactic acid production with a new acid hydrolysis of ram horn waste. AB - The use of ram horn hydrolysate (RHH) as a substrate for lactic acid production was investigated using Lactobacillus casei. For this purpose, first RHH was produced. Ram horns were hydrolyzed by treating with acids (6N-H2SO4 and 6N-HCl) and neutralizing the solutions. The amounts of protein, nitrogen, ash, some minerals, total sugars, total lipids and amino acids of the RHH were determined. The effect of different concentrations (1-10% v/v) of RHH on the production of biomass, lactic acid concentration and sugar consumption was investigated, and a concentration of 6% RHH was found to be optimal. The content of lactic acid in the culture broth containing 6% RHH (44 g x l(-1)) for 26 h was 22% higher than that of the control culture broth (36 g x l(-1)). From this result, RHH was demonstrated to be a suitable supplement for lactic acid production, a use that would resolve a local environmental problem. PMID- 12900018 TI - BldD from Streptomyces coelicolor is a non-essential global regulator that binds its own promoter as a dimer. AB - We have shown that the bldD gene of Streptomyces coelicolor, while required for antibiotic production and morphological differentiation, is not essential for viability. We have also demonstrated that BldD forms a higher order complex both in solution and when bound to target DNA. Purified BldD exists in three forms in solution, as a tetramer, dimer and monomer, but only in the dimeric form when bound to its own promoter/operator. PMID- 12900020 TI - Pythium glomeratum, a new species isolated from agricultural soil taken in north eastern France, its ITS region and its comparison with related species. AB - Pythium glomeratum sp. nov. is described here. It was isolated from soil samples taken in the northern France in 1992, was wrongly identified as Pythium heterothallicum and was kept aside. Recently, the ITS region of the rDNA of this oomycete was amplified and sequenced. The differences between the sequence and a more detailed study of the morphological characters of these two species, revealed that both are related but different species. P. glomeratum is characterized by the presence of branched antheridia that wrap around the oogonia, aplerotic to almost plerotic oospores, and the lack of zoosporangia and zoospores. Taxonomical description of this new species, its comparison with related oomycetes, polymerase chain reaction (PCR) amplification of the internal transcribed region (spacers ITS1, ITS2, and the gene 5.8S) of its ribosomal nuclear DNA and the nucleotide sequence of this region are given here. PMID- 12900019 TI - Expression and localization of the linear DNA plasmid-encoded protein (RS224) in Rhizoctonia solani AG2-2. AB - Expression of the linear DNA plasmid-encoded protein (RS224) from the plant pathogenic fungus Rhizoctonia solani isolate H-16, anastomosis group 2-2, and its localization were studied. Extracts from Escherichia coli cells expressing the open reading frame (ORF) of RS224 (RS224ORF in pRS224) contain a 92-kDa T7.Tag RS224orf fusion protein. Antisera raised against the fusion protein obtained from E. coli cells cross-reacted with a 90-kDa protein in the mycelia. To analyze the subcellular localization of the 92-kDa protein, mycelia of R. solani were disrupted and fractionated. Antibodies against RS224 proteins specifically reacted to the mitochondrial fraction, suggesting that RS224 is localized in mitochondria. PMID- 12900021 TI - A gene transfer system for the glycopeptide producer Nonomuraea sp. ATCC39727. AB - The filamentous actinomycete Nonomuraea sp. ATCC39727 produces the industrially important glycopeptide antibiotic A40926. We developed a gene transfer system based on intergeneric conjugation from Escherichia coli. Analysis of the ex conjugants revealed that the incoming plasmid pSET152 had integrated at two sites in the Nonomuraea genome. One of these was characterized and found to be highly related to other PhiC31 attB sites described in Streptomyces spp., including the core TTS sequence, where crossover occurs. Surprisingly, pSET152 was also found in episomic form in the Nonomuraea ex-conjugants. PMID- 12900022 TI - The Streptomyces coelicolor ssgB gene is required for early stages of sporulation. AB - ssgB was identified as a novel early sporulation gene in Streptomyces coelicolor. An ssgB deletion mutant failed to sporulate, over-produced actinorhodin, and its colonies were significantly larger than those of the parental strain, suggesting an important role for the ssgB gene product in the process of growth cessation prior to sporulation-specific cell division. This places ssgB temporally before the paralogous sporulation gene ssgA. Analysis of ssgB mutant hyphae by electron microscopy and by confocal fluorescence microscopy showed that it was defective in the initiation of sporulation, as no sporulation septa could be identified, and DNA segregation had not yet been initiated in the mutant. PMID- 12900023 TI - Construction of a genetically engineered microorganism for CO2 fixation using a Rhodopseudomonas/Escherichia coli shuttle vector. AB - The CO2 fixation ability of Rhodopseudomonas palustris DH was enhanced by introducing the recombinant plasmid pMG-CBBM containing the form II ribulose-1,5 bisphosphate carboxylase/oxygenase (RubisCO) gene (cbbM) isolated from Rps. palustris NO. 7. Sequencing of a 3.0-kb PstI fragment containing the cbbM gene revealed an open reading frame encoding 461 amino acids, homologous to known cbbM genes, with a ribosome binding site upstream of cbbM and a terminator downstream of cbbM, without promoter. pMG-CBBM, a Rhodopseudomonas/Escherichia coli shuttle expression plasmid, was derived from the Rhodopseudomonas/E. coli shuttle cloning vector pMG105, by inserting the promoter of the pckA gene and the cbbM gene into its multiple cloning site. Plasmid pMG-CBBM was transformed into Rps. palustris DH by electroporation, and was stably maintained when transformants were grown either photoheterotrophically or photolithoautotrophically in the absence of antibiotics. This is the first report of an expression plasmid containing a Rps. palustris-specific promoter that allows stable expression of a foreign gene in the absence of antibiotic selection. PMID- 12900024 TI - Role of 5'-TGN-3' motif in the interaction of mycobacterial RNA polymerase with a promoter of 'extended -10' class. AB - In a systematic approach to understand the transcriptional machinery of mycobacteria, we had previously isolated and characterized mycobacterial promoter regions. In this study, we have investigated molecular interactions between mycobacterial RNA polymerase holoenzyme, reconstituted with different sigma subunits and the promoter element of the Mycobacterium tuberculosis gene pknH (Rv1266c), a representative of promoters belonging to the 'extended -10' class. In vitro transcription assays using the pknH promoter and reconstituted RNA polymerase holoenzyme demonstrated that transcription from the pknH promoter is specifically initiated by sigmaA, the principal sigma factor of mycobacteria. DNase I protection assay and deletion studies with the pknH promoter revealed that the minimal region required for optimal transcription carries the sequence from position -37 to position +6. Moreover, mutation in the TGN motif of the pknH promoter resulted in the loss of >75% of its activity. Binding of RNA polymerase with wild-type promoter as well as its TG- mutant revealed that the TGN motif is required for the transition from a close complex into an open complex. Further, it was observed that the presence of the TGN motif reduces the thermal energy required for the conversion of a close complex into an open complex, necessary for initiation of transcription. PMID- 12900025 TI - Cloning and expression of the inorganic pyrophosphatase gene from the amino acid producer Brevibacterium lactofermentum ATCC 13869. AB - A 20-kDa Brevibacterium lactofermentum protein was detected when purifying the His-tagged FtsZBL. The protein was identified by matrix-assisted laser desorption/ionisation time of flight as the inorganic pyrophosphatase encoded by the ppa gene, which is present as a single copy in the genome of Corynebacterium glutamicum. The ppa gene was cloned from B. lactofermentum chromosomal DNA by polymerase chain reaction; it seemed to be an essential gene and it might represent an attractive target for drug discovery. The cloned ppa gene complemented a ppa- Escherichia coli mutant and a ppa-gfp gene fusion revealed that the gene product mainly accumulated at the cell poles in both E. coli and B. lactofermentum. PMID- 12900026 TI - The putative anti-anti-sigma factor BldG is post-translationally modified by phosphorylation in Streptomyces coelicolor. AB - The Streptomyces coelicolor bldG gene encodes a protein showing similarity to the SpoIIAA and RsbV anti-anti-sigma factors of Bacillus subtilis. Purified maltose binding protein-BldG could be phosphorylated in vitro by wild-type S. coelicolor crude extract, and both the phosphorylated and unphosphorylated forms of BldG could be detected in vivo using isoelectric focusing. ATP was shown to serve as the phosphoryl group donor, and phosphorylation of BldG was abolished when the putative phosphorylation site was changed from a serine to an alanine residue. A bldG mutant strain expressing the non-phosphorylatable BldG protein was unable to undergo morphological differentiation or produce antibiotics even after prolonged incubation, suggesting that phosphorylation of BldG is necessary for proper development in S. coelicolor. PMID- 12900028 TI - Systematic characterization of Escherichia coli genes/ORFs affecting biofilm formation. AB - To understand the nature and function of bacterial biofilm and the process of its formation, we have performed systematic screening of a complete set of Escherichia coli genes/open reading frames (ORFs) to identify those that affect biofilm development upon over-expression. In contrast to the biofilm of strain AG1 used as a control, some of the genes/ORFs when over-expressed led to the formation of an abnormal biofilm such as thin, mat-like, filamentous or one easily detaching from various surfaces. Disruptants of selected genes were constructed in order to clarify their roles in the different stages of biofilm formation. Our results suggest that diverse metabolic pathways contribute to the development of biofilm. PMID- 12900027 TI - Construction and characterization of a Porphyromonas gingivalis htpG disruption mutant. AB - Our previous reports implicated the Hsp90 homologue (HtpG) of Porphyromonas gingivalis (Pg) in its virulence in periodontal disease. We investigated the role of the HtpG stress protein in the virulence of Pg. This report describes the (i) expression of a recombinant Pg HtpG (rHtpG), (ii) generation and characterization of a polyclonal rabbit anti-Pg rHtpG antiserum, and (iii) construction of a Pg htpG isogenic mutant and evaluation of the growth, adherence and invasion properties compared to the wild-type parental strain. The disruption of the htpG gene did not significantly affect growth, and had no effect on Pg adherence to and invasion of cultured human cells. PMID- 12900029 TI - Development of specific fluorescent oligonucleotide probes for in situ identification of wine lactic acid bacteria. AB - A rapid method for the identification of lactic acid bacteria (LAB) from wine has been developed. This method is based on fluorescence in situ hybridisation (FISH), using fluorescent oligonucleotide probes, homologous to 16S rDNA of those species of LAB commonly found in wines. The protocol for the specific detection of these bacteria was established through the hybridisation of 36 reference strains. The specificity of the probes was evaluated by using pure cultures. Probes were used to identify species in different wines, making it evident that direct identification and quantification from natural samples without culturing is also possible. The results show that FISH is a promising technique for the rapid identification of LAB, allowing positive identification in a few hours (4 16 h). PMID- 12900030 TI - In vitro conjugal transfer of tetracycline resistance from Lactobacillus isolates to other Gram-positive bacteria. AB - The ability of 14 Lactobacillus strains, isolated from fermented dry sausages, to transfer tetracycline resistance encoded by tet(M) through conjugation was examined using filter mating experiments. Seven out of 14 tetracycline-resistant Lactobacillus isolates were able to transfer in vitro this resistance to Enterococcus faecalis at frequencies ranging from 10(-4) to 10(-6) transconjugants per recipient. Two of these strains could also transfer their resistance to Lactococcus lactis subsp. lactis, whereas no conjugal transfer to a Staphylococcus aureus recipient was found. These data suggest that meat lactobacilli might be reservoir organisms for acquired resistance genes that can be spread to other lactic acid bacteria. In order to assess the risk of this potential hazard, the magnitude of transfer along the food chain merits further research. PMID- 12900031 TI - Extragenic suppression of the requirement for diaminopimelate in diaminopimelate auxotrophs of Mycobacterium smegmatis. AB - Mycobacteria, like many prokaryotes, have a peptidoglycan with peptides composed of L-alanine (or glycine), D-iso-glutamine, meso-diaminopimelate, and D-alanine. We sought to study mycobacterial peptidoglycan biosynthesis by constructing diaminopimelate (DAP) auxotrophs of Mycobacterium smegmatis and then isolating spontaneous mutants of these auxotrophs that can grow in the absence of DAP. Here we report the isolation and characterization of seven classes of spontaneous M. smegmatis mutants with extragenic mutations that can suppress the DAP requirement of DAP auxotrophs. PMID- 12900032 TI - Conserved temperature-dependent expression of RNA-binding proteins in cyanobacteria with different temperature optima. AB - The expression of the rbp genes, which encode small RNA-binding proteins with a single RNA-recognition motif, is known to increase at low temperature in Anabaena variabilis M3. The 5'-untranslated region (UTR) of the rbpA1 gene is involved in the cold-regulation. We compared the regulation of the rbp genes in three strains of cyanobacteria having different temperature optima, namely, a mesophilic strain Anabaena sp. PCC 7120, a thermophilic strain Thermosynechococcus elongatus BP-1, and a psychrophilic Antarctic strain Oscillatoria sp. SU1. In Anabaena 7120 and T. elongatus, all the rbp gene sequences are known, and the 5'-UTR sequences of some rbp genes have a high similarity to the 5'-UTR of rbpA1. We found that transcripts as well as protein products of these rbp genes accumulated at low temperature. In addition, the expression of rbp genes increased at low temperature in the Oscillatoria sp. SU1. This suggests that a mechanism of cold regulation of rbp genes is common among various species of cyanobacteria that belong to different taxa and have different temperature optima. PMID- 12900033 TI - Presence of yeasts in southern Italian sourdoughs from Triticum aestivum flour. AB - Samples of sourdoughs obtained from 13 artisanal bakeries located in the Molise and Campania regions were analysed. The sourdoughs were produced with the exclusive use of Triticum aestivum wheat flour. pH values of sourdoughs from Molise were generally lower than those from Campania. The number of yeasts in the samples of sourdoughs from Molise was generally higher than in those from Campania, which in two cases evidenced counts about 2 log cfu x g(-1). By utilising and comparing traditional and biomolecular techniques of identification a complete picture of the isolates was obtained: 58 strains were identified as Saccharomyces cerevisiae, five as Candida colliculosa, four as C. lambica, three as C. krusei, three as C. valida and two as C. glabrata. PMID- 12900035 TI - Identification of the promoter regions and sigma(s)-dependent regulation of the gadA and gadBC genes associated with glutamate-dependent acid resistance in Shigella flexneri. AB - Resistance to killing by low pH is a common feature of both Escherichia coli and Shigella flexneri. The most effective E. coli acid resistance system utilizes two isoforms of glutamate decarboxylase encoded by gadA and gadB, and a putative glutamate/gamma-amino butyric acid antiporter encoded by gadC. Expression of the gad system is dependent upon the alternate sigma factor, sigma(s). We confirm that gadA, gadB, and gadC are also all dependent upon sigma(s) for their expression in S. flexneri. -10 sequences similar to the sigma(s)-10 consensus sequence were identified by primer extension in the upstream promoters of all three genes and the transcriptional start points were identical in both E. coli and S. flexneri. PMID- 12900034 TI - Characterization of In111, a class 1 integron that carries the extended-spectrum beta-lactamase gene blaIBC-1. AB - A class 1 integron, In111, carried by a self-transferable plasmid from an Escherichia coli clinical strain was characterized. The variable region of In111 constituted an array of gene cassettes encoding the extended-spectrum beta lactamase IBC-1, the aminoglycoside-modifying enzymes AAC(6')-Ib and ANT(3")-Ia, dihydrofolate reductase I and a putative polypeptide (SMR-2) sharing similarity with the Qac transporters. Transcription of the gene cassettes was driven by a hybrid-type P1 promoter located in a typical 5' conserved segment (CS). The 3'CS included sulI, qacEDelta1, orf5 and orf6. In111 was bounded on the right by an inversely oriented IRt. The 5'CS was preceded by an intact IS26 element followed by an aphA1 gene. PMID- 12900036 TI - Chloride dependence of growth in bacteria. AB - Chloride is an abundant anion on earth but studies analyzing a possible function of chloride in prokaryotes are scarce. To address the question, we have tested 44 different Gram-negative and Gram-positive bacteria for a chloride dependence or chloride stimulation of growth. None required chloride for growth at their optimal growth (salt) conditions. However, in hyperosmotic media containing high concentrations of Na+, 11 bacteria (Aeromonas hydrophila, Bacillus megaterium, Bacillus subtilis, Corynebacterium glutamicum, Escherichia coli, Paracoccus denitrificans, Proteus mirabilis, Proteus vulgaris, Staphylococcus aureus, Thermus thermophilus, and Vibrio fischeri) had a strict chloride dependence for growth or were significantly stimulated by chloride. These data show that chloride is essential for growth at high salt (Na+) concentrations in various species of the domain Bacteria. PMID- 12900037 TI - Oxidant-inducible resistance to hydrogen peroxide killing in Agrobacterium tumefaciens requires the global peroxide sensor-regulator OxyR and KatA. AB - Induced adaptive and cross-protective responses to peroxide stress are important strategies used by bacteria to survive stressful environments. We have shown that exposure to low levels of peroxide (adaptive) and superoxide anions (cross protection) induced high levels of resistance to peroxide killing in Agrobacterium tumefaciens. The mechanisms and genes involved in these processes have not been identified. Here, the roles played by peroxide (oxyR) and superoxide (soxR) global regulators and a catalase gene (katA) during these responses were investigated. H2O2-induced adaptive protection was completely abolished in both the oxyR and katA mutants. Superoxide generator (menadione) induced cross-protection to H2O2 killing was observed in a soxR mutant, but not in either an oxyR or a katA mutant. In vivo analysis of the katA promoter, using a katA::lacZ transcriptional fusion, revealed that it could be induced by menadione in an oxyR-dependent manner. These results lead us to conclude that H2O2 and superoxide anions directly or indirectly oxidize OxyR and it is the resulting activation of katA expression that is responsible for the induced protection against lethal concentrations of H2O2. PMID- 12900038 TI - Spatial and non-spatial learning in turtles: the role of medial cortex. AB - In mammals and birds, hippocampal processing is crucial for allocentric spatial learning. In these vertebrate groups, lesions to the hippocampal formation produce selective impairments in spatial tasks that require the encoding of relationships among environmental features, but not in tasks that require the approach to a single cue or simple non-spatial discriminations. In reptiles, a great deal of anatomical evidence indicates that the medial cortex (MC) could be homologous to the hippocampus of mammals and birds; however, few studies have examined the functional role of this structure in relation to learning and memory processes. The aim of this work was to study how the MC lesions affect spatial strategies. Results of Experiment 1 showed that the MC lesion impaired the performance in animals pre-operatively trained in a place task, and although these animals were able to learn the same task after surgery, probe test revealed that learning strategies used by MC lesioned turtles were different to that observed in sham animals. Experiment 2 showed that the MC lesion did not impair the retention of the pre-operatively learned task when a single intramaze visual cue identified the goal. These results suggest that the reptilian MC and hippocampus of mammals and birds function in quite similar ways, not only in relation to those spatial functions that are impaired, but also in relation to those learning processes that are not affected. PMID- 12900039 TI - Voluntary control of postural equilibrium patterns. AB - The ability to voluntarily transit from one whole-body movement to another is based on the multisensory integration of visual, vestibular, and somatosensory information. The role of functional sensory ranges and mechanical constraints on the ability to voluntarily transit between whole-body movements was studied by requiring subjects to switch from a head-fixed-to-surface to head-fixed-in-space postural pattern (and vice versa). The head-fixed-to-surface pattern required an erect stance characterized by an in-phase relationship between center of pressure (CoP) and platform motion. The head-fixed-in-space pattern required subjects to fix trunk-head position in-space while producing an anti-phase relationship between CoP and platform motion. The voluntary transition was performed with and without vision while standing on a surface oscillating in the anterior-posterior (A/P) direction. The support surface oscillated at five frequencies (0.2-1Hz) with amplitude fixed at 15cm. The voluntary transition was initiated with an auditory cue. The appropriate CoP-platform phase relationship for the two postural patterns was produced for all frequencies with and without vision. Upper trunk kinematics revealed that subjects often failed to produce the head-fixed-to surface pattern for frequencies >/=0.6Hz, while producing the head-fixed-in-space pattern at all frequencies with vision. Without vision, neither pattern was produced consistently based on upper-trunk kinematics. These findings demonstrate separate control processes for upper- and lower-body motion and that functional sensory ranges and mechanical constraints can facilitate or inhibit voluntary production of whole-body movements based on these control processes. The results are discussed in reference to neurological substrates that may be involved in the planning and execution of motor set-switching. The experimental protocol we employ may also have application as a diagnostic tool for the evaluation of postural deficits. PMID- 12900040 TI - Ictal stimulus processing during spike-wave discharges in genetic epileptic rats. AB - In the present experiment it was investigated whether and to what extent auditory information processing is possible during the presence of spike-wave discharges in rats. To that end, WAG/Rij rats which are an animal model for absence epilepsy, were provided with cortical electrodes for the registration of the electroencephalogram (EEG). The animals were first trained in an appetitively motivated conditioning paradigm to learn to discriminate between two auditory stimuli with equal duration and frequency but with different intensities. Next, the stimuli were presented in the test phase in pseudorandom order during spike wave discharges. The reactivity of the ongoing EEG was analysed. It was found that the presentation of the reinforced stimulus induced a larger number of aborted spike-wave discharges than the non-reinforced stimulus, regardless of the intensity of the stimuli. This implies that during generalised spike-wave discharges the brain is still capable of evaluating the meaning of an ictally presented stimulus. It also shows that sensory, attentional and mnemonic processes are at least partially intact during the occurrence of a spike-wave discharge. The results of the present study are largely in agreement with results on human spike-wave activity-related cognitive disturbances. Moreover, they may lead to a refinement of the concept of epileptic consciousness and may emphasise the heuristic value of rodent models for studying both ictal and interictal information processing. PMID- 12900041 TI - Infusion of apomorphine into the dorsocentral striatum produces acute drug induced recovery from neglect produced by unilateral medial agranular cortex lesions in rats. AB - Previous studies have shown that systemic administration of apomorphine is effective in producing acute drug-induced recovery from neglect induced by unilateral medial agranular cortex (AGm) lesions. More recent studies have demonstrated that recovery from neglect may be due to plastic changes occurring in the dorsal central striatum (DCS). Further, lesions of the DCS produce neglect that does not respond to systemic administration of apomorphine, suggesting that this area may be crucial for the therapeutic effects of apomorphine. In the present study, the behavioral effects of apomorphine infused into the DCS of animals with AGm lesion-induced neglect were examined to determine whether the DCS is a site of drug action. An infusion of 0.375 micro g of apomorphine into the DCS, but not a lateral striatal control area, was effective in producing acute recovery from neglect. The results of this study support the crucial role of the DCS in recovery from neglect induced by unilateral AGm lesions and suggest that the DCS may be an important site of action for the therapeutic effects of apomorphine. Because dopamine agonist therapy has been shown to be effective in humans with neglect, the results of the current study may represent an important step in the development of future pharmacotherapies. PMID- 12900042 TI - Mecamylamine reversal by nicotine and by a partial alpha7 nicotinic acetylcholine receptor agonist (GTS-21) in rabbits tested with delay eyeblink classical conditioning. AB - The aim of this experiment was to investigate the effects of nicotinic acetylcholine receptor (nAChR) agonism and antagonism on learning. Eyeblink classical conditioning (750ms delay procedure) was tested for 15 daily sessions in a total of 82 young rabbits: 58 rabbits were tested in the paired procedure when the conditioned stimulus (CS) was always followed by the unconditioned stimulus (US), and 24 rabbits were tested in the explicitly unpaired procedure in which CS and US presentations were independent. We used the nAChR agonists nicotine and GTS-21 (a selective alpha7 nAChR partial agonist that antagonizes alpha4beta2 nAChRs) and the relatively nonselective nAChR antagonist, mecamylamine. Groups of young rabbits were injected with 0.5mg/kg mecamylamine alone and in combination with two doses of nicotine or GTS-21 and compared to vehicle-treated rabbits. Explicitly unpaired control groups received vehicle, mecamylamine plus the highest nicotine dose, or mecamylamine plus the highest GTS 21 dose. Both GTS-21 and nicotine reversed the deleterious effect of mecamylamine on the acquisition of conditioned responses. Combinations of GTS-21 or nicotine and mecamylamine did not cause sensitization or habituation in the unpaired condition. Reversal of mecamylamine-induced learning deficits by nicotine and GTS 21 suggests that nAChR agonists may have efficacy in ameliorating deficits caused by the loss of some types of nAChRs in diseases such as AD. PMID- 12900043 TI - Effect of dorsal periaqueductal gray lesion on cardiovascular and behavioural responses to contextual conditioned fear in rats. AB - Contextual conditioned fear in the rat is characterized by a freezing immobility associated with a marked increase in blood pressure, a slow increase in heart rate, and ultrasonic vocalizations. A previous Fos study also revealed a marked activation of the ventrolateral part of the periaqueductal gray (VLPAG) and a much smaller activation of its dorsal part (DPAG). Recent chemical blockade experiments indicate that the main role of the VLPAG in the response is to impose the immobility necessary for the expression of the freezing component. We now test the role of the DPAG to see if its small activation (as revealed by Fos) is of any functional significance in the contextual fear response. Large N-methyl-D aspartate (NMDA) excitotoxic lesions that destroyed most of the DPAG were made in 10 rats. Another group of 10 rats had sham lesions with saline. The animals were then implanted with blood pressure telemetric probes, fear conditioned, and finally tested. There was no significant difference in the amount of freezing and in the blood pressure response between the two groups. However, there was a complete abolition of ultrasonic vocalizations and a significantly greater increase in heart rate in the DPAG-lesioned group. The effect on vocalization and heart rate may be explained by lesion of adjacent structures: the lateral PAG and the superior colliculus (baroreflex alteration), respectively. Thus, most of DPAG appears to play little role in the expression of the contextual fear response. PMID- 12900044 TI - Transplantation of neurospheres after granule cell lesions in rats: cognitive improvements despite no long-term immunodetection of grafted cells. AB - EGF-responsive C17 murine-derived neural stem cells (neurospheres) were grafted into the dentate gyrus of adult male rats after dentate granule cells lesions produced by colchicine injections. Behavioural performance was evaluated over two post-grafting periods, using tests sensitive to hippocampal dysfunctions. The first period began 1 month after grafting and testing conducted in the water maze and the radial maze distinguished working- and reference-memory performance. The second period began 9 months after grafting and learning performance was also evaluated in a Hebb-Williams maze, in addition to both other tests. The lesions induced lasting deficits in all tests. During the first period, the grafts had no effect in either test. Conversely, during the second period, grafted rats showed a weak improvement in the water maze and a significant increase of reference memory performance in the radial maze. In the Hebb-Williams maze, performance of grafted rats was close to normal. Strengthening the idea that dentate gyrus granule cells play an important role in the acquisition of new (perhaps more configural than only spatial) information, our results, moreover, suggest that neurosphere grafts may foster recovery after damage to point-to-point connection systems in the adult brain. PMID- 12900045 TI - Antidepressant-like effects in various mice strains in the tail suspension test. AB - Several studies have reported rodent strain differences in the response to antidepressants in animal models of depression. The aim of the present study was to investigate the potential contribution of genetic factors to antidepressant response in an animal model of depression: the tail suspension test (TST). For this study four mice strains (Swiss and NMRI, two outbred strains and DBA/2 and C57BL/6J Rj, two inbred strains) were submitted to the TST after acute administration of five antidepressants: the tricyclic antidepressants (TCAs) imipramine and desipramine, the selective serotonin (5-HT) reuptake inhibitors (SSRIs) paroxetine and citalopram and the dopamine reuptake inhibitor bupropion. The C57BL/6J Rj strain had a longer baseline immobility time in comparison to the other strains. All antidepressants studied in this work decreased immobility time in the Swiss and C57BL/6J Rj strains. However, the Swiss strain displayed greater sensitivity to citalopram (from 2mg/kg) and C57BL/6J Rj to paroxetine (from 0.5mg/kg). This latter presented a greater size-effect with citalopram than with other strains and reached more than 60% from 8mg/kg. Moreover the size-effect of desipramine, paroxetine and bupropion in Swiss mice was greater than in the other strains in the TST. The NMRI and DBA/2 mice only responded to 5-HT reuptake inhibitors, both selective (paroxetine, citalopram) or non-selective (imipramine). The NMRI strain was more sensitive to imipramine and presented a size-effect (43% at 8mg/kg) superior to those of other strains. DBA/2 strain was more sensitive to citalopram than paroxetine and imipramine. Our results suggest that response to an antidepressant treatment is under control of genetic factors and that the strain of mouse is an important parameter to consider. PMID- 12900047 TI - Perinatal hypothyroidism effects on neuromotor competence, novelty-directed exploratory and anxiety-related behaviour and learning in rats. AB - Thyroid hormone is essential for proper development of the mammalian CNS. Previous studies have documented a decrease in the ability of neonatal hypothyroid animals to learn and to habituate to maze tests and an increase in spontaneous activity. However, there is little information about the effects of perinatal (i.e. perinatal and postnatal) hypothyroidism on behaviour. The aim of the present work was to investigate the longitudinal effects of perinatal hypothyroidism on certain aspects of the behaviour in rats. Neuromotor competence was tested at 21, 40 and 60 days, novelty-directed exploratory behaviour and anxiety-related behaviour were evaluated at 40 and 60 days by means of the Boissier tests and associative learning ability was tested at 80 days by means of a step-through passive avoidance task. The persistence of the effects of perinatal hypothyroidism on psychomotor performance was highly dependent on the task examined. Perinatal hypothyroidism caused an increase of locomotor activity as revealed by the total distance travelled in the Boissier test and this increase also comprised a component of decreased anxiety-related behaviour. Methimazole-treated subjects also had higher head-dip scores than controls at 40 days while no differences were observed at 60 days. Finally, our results showed that methimazole-treated rats performed poorly in a passive avoidance learning task. PMID- 12900046 TI - Reduced startle habituation and prepulse inhibition in mice lacking the adenosine A2A receptor. AB - Adenosine and dopamine receptors interact in the CNS to modulate behaviour, including sensorimotor gating. Prepulse inhibition (PPI) has been suggested to be an operational measure of sensorimotor gating. PPI and startle habituation are disrupted in patients with schizophrenia. In experimental animals, both parameters are modulated by dopaminergic and adenosine receptor agonists and antagonists. In the present study, we measured PPI and startle habituation in mice that lack functional adenosine A(2A) receptors. Startle amplitudes, startle habituation and PPI were significantly reduced in mice homozygous null for the adenosine A(2A) receptor (A(2A)(-/-)). In addition, differential effects of amphetamine and MK-801 on startle amplitude, startle habituation and PPI were observed between A(2A)(-/-) and wildtype controls. These data support the involvement of adenosine A(2A) receptors in regulation of PPI and startle habituation. PMID- 12900048 TI - Novel vaccine strategies with protein antigens of Streptococcus pneumoniae. AB - Infections caused by Streptococcus pneumoniae (pneumococcus) are a major cause of mortality throughout the world. This organism is primarily a commensal in the upper respiratory tract of humans, but can cause pneumonia in high-risk persons and disseminate from the lungs by invasion of the bloodstream. Currently, prevention of pneumococcal infections is by immunization with vaccines which contain capsular polysaccharides from the most common serotypes causing invasive disease. However, there are more than 90 antigenically distinct serotypes and there is concern that serotypes not included in the vaccines may become more prevalent in the face of continued use of polysaccharide vaccines. Also, certain high-risk groups have poor immunological responses to some of the polysaccharides in the vaccine formulations. Protein antigens that are conserved across all capsular serotypes would induce more effective and durable humoral immune responses and could potentially protect against all clinically relevant pneumococcal capsular types. This review provides a summary of work on pneumococcal proteins that are being investigated as components for future generations of improved pneumococcal vaccines. PMID- 12900049 TI - Immunobiotics and the probiotic evolution. AB - The term 'probiotic' was coined by the food industry to describe microbes beneficial to health. Scientific criticism based on poorly characterised isolates, soft clinical targets, and an absence of a mechanistic framework limited value of this concept. Current clinical and animal model studies support a role for certain 'probiotics', activating the common mucosal system through the stimulation of gut antigen-presenting cells to both promote protection and to switch regulatory mechanisms. It is concluded that a new term is required to identify bacteria that promote health through driving mucosal immune mechanisms, compared to those with strictly local effects. The term 'immunobiotics' is suggested as appropriate to fulfil this need. PMID- 12900050 TI - Regulation of innate and adaptive immunity by the female sex hormones oestradiol and progesterone. AB - Women mount more vigorous antibody- and cell-mediated immune responses following either infection or vaccination than men. The incidence of most autoimmune diseases is also higher in women than in men; however, during pregnancy many autoimmune diseases go into remission, only to flare again in the early post partum period. Successful pregnancy requires that the female immune system tolerate the presence of a semi-allogeneic graft for 9 months. Oral contraceptive use can increase susceptibility to certain genital tract infections and sexually transmitted diseases in women. Moreover, treatment of mice and rats with female sex hormones is required to establish animal models of genital tract Chlamydia, Neisseria and Mycoplasma infection. This review describes what is currently known about the effects of the female sex hormones oestradiol and progesterone on innate and adaptive immune responses in order to provide a framework for understanding these sex differences. Data from both human and animal studies will be reviewed. PMID- 12900051 TI - Loss of hemolysin expression in Staphylococcus aureus agr mutants correlates with selective survival during mixed infections in murine abscesses and wounds. AB - During the screening of a Staphylococcus aureus signature-tagged mutagenesis library, it was noted that nonhemolytic bacteria became more abundant as time passed in murine abscess and wound models, but not within organ tissues associated with systemic infections. To examine this further, a mixed population of hyperhemolytic, hemolytic, and nonhemolytic S. aureus strain RN6390 cells were inoculated into mice using abscess, wound, and systemic models of infection. After 7 days in the abscess, the hyperhemolytic group markedly declined, whereas the nonhemolytic population increased significantly. A similar phenomenon occurred in murine wounds, but not during the systemic infection. Sequencing of several of the signature-tagged mutants indicated mutations in the agrC gene or within the agrA-agrC intergenic region. Both alpha-hemolysin and delta-hemolysin activity was curtailed in these mutants, but beta-hemolysin activity was unaffected. Single strain comparisons between wild-type strain 8325-4 and strain DU1090 (hla-) as well as between strain RN6911 (agr) and wild-type strain RN6390 were performed using the same three animal models of infection. The agr mutant strain and the hla mutant strain showed no difference in bacterial counts in murine wounds compared to their respective parent strains. The same held true in murine abscesses at day 4, but strain RN6911 counts then declined at day 7. Considerable clearing of the hla mutant strain and the agr mutant strain occurred in the systemic model of infection. Mixed infections with the DU1090 and 8325-4 strains in the abscess model showed a slight advantage given to the DU1090 population, but a distinct selection for the parental 8325-4 strain in the liver. These results suggest that agr mutations cause reductions in the expression of several secreted proteins, including alpha- and delta-hemolysin, which in turn contribute to a growth advantage of this agr mutant group within a mixed population of S. aureus cells residing in abscesses and wounds. PMID- 12900052 TI - Recombinant expression and antigenic properties of a 31.5-kDa keratinolytic subtilisin-like serine protease from Microsporum canis. AB - A secreted 31.5-kDa keratinolytic subtilase (SUB3; AJ431180) is thought to be a Microsporum canis virulence factor and represents a candidate for vaccination trials. In this study, the recombinant keratinase (r-SUB3) was produced by the Pichia pastoris expression system and purified to homogeneity. Recombinant SUB3 displayed identical biochemical properties with the native protease. Experimentally cutaneously infected guinea pigs showed specific lymphoproliferative response towards r-SUB3, while no specific humoral immune response was induced except for one animal. The heterologous expression of SUB3 provides a valuable tool for addressing further investigations on the role of this keratinase in the specific cellular immune response and on its use in vaccination trials in the cat. PMID- 12900053 TI - Identification, purification and characterization of a receptor for dengue virus induced macrophage cytotoxin (CF2) from murine T cells. AB - Dengue type-2 virus infection in mice induces a subpopulation of T lymphocytes to produce a cytokine cytotoxic factor, which induces macrophages (Mphi) to produce a biologically active cytotoxic cytokine, the Mphi cytotoxin (CF2). Previously we have identified the presence of intermediate-affinity receptors for CF2 on mouse peritoneal Mphi. The present study was undertaken to identify the CF2-receptors (CF2-R) on murine T cells followed by their purification and characterization. Receptor binding assay and Scatchard analysis revealed single, high-affinity (1.0309 nM) receptors for CF2 on T cells (22000 receptors per cell). The binding of [125I]CF2 on murine T cells was saturable and specific. Furthermore, CF2-R was purified from normal mouse T cell plasma membrane by affinity chromatography followed by reversed-phase high-pressure liquid chromatography. The presence of CF2-R was confirmed by indirect dot-blot assay and its binding with [125I]CF2. The purified CF2-R is a 90-95-kDa protein as characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblot analysis. The chemical crosslinking of [125I]CF2 and its receptor complex showed a product of 100-110 kDa on different subpopulations of murine T cells. The pretreatment of target cells with anti-CF2-R antisera inhibited the cytotoxic activity of CF2 in a dose dependent manner and thus confirmed the biological significance of CF2-R. Moreover, the presence of CF2-R was also identified on normal human peripheral blood mononuclear cells and T and B cells by crosslinking with [125I]CF2, thus revealing the possible role of CF2 and CF2-R in the immunopathogenesis of dengue virus disease. PMID- 12900054 TI - The stress-induced hsp12 gene shows genetic variation among Helicobacter pylori strains. AB - The transcriptional regulation, genetic variation and clinical relevance of the strain-specific hsp12 gene of the human gastric pathogen Helicobacter pylori were investigated. Although the transcription of the hsp12 gene in H. pylori strain 1061 was induced by growth under iron-, pH- and temperature-stress conditions, the gene was not essential for growth under these stress conditions. The locus containing the hsp12 gene showed considerable genetic variation. A total of eight different strain-specific alleles were identified, of which three are mosaic variants of the hsp12 gene and five that are unrelated to the hsp12 gene. The hsp12 locus of six paired sets of strains obtained from patients with 7-10-year time intervals remained unaltered, indicating that genetic variation does not occur during chronic infection. No significant association was found between the presence of a hsp12 gene and peptic ulcer disease in clinical isolates obtained from 26 patients. The stress-regulated, strain-specific hsp12 genes may be involved in adaptation of individual H. pylori strains to their specific hosts, and contribute to long-term colonization of the gastric niche. PMID- 12900055 TI - Serum high-density lipoprotein (HDL) inhibits in vitro enterohemolysin (EHly) activity produced by enteropathogenic Escherichia coli. AB - Enterohemolysin (EHly) produced by Escherichia coli shows hemolytic activity towards washed erythrocytes from different animal species on blood agar plates. It has been shown recently that EHly activity is inhibited by normal mammalian serum and by cholesterol in vitro. Plasma lipoproteins can interact with bacterial toxins, such as endotoxin, to reduce their toxicity. In this work, we examine the ability of human purified chylomicrons, very low-density lipoproteins, intermediate-density, low-density and high-density lipoproteins, to inhibit the hemolytic activity of EHly. Our results show that these lipoproteins are hemolysin inactivators, and that high-density lipoprotein is the most potent inhibitor of enterohemolytic activity. PMID- 12900056 TI - Rhinovirus increases human beta-defensin-2 and -3 mRNA expression in cultured bronchial epithelial cells. AB - Human beta-defensins (hBDs) are antimicrobial peptides that play important roles in host defense against infection, inflammation and immunity. Previous studies showed that micro-organisms and proinflammatory mediators regulate the expression of these peptides in airway epithelial cells. The aim of the present study was to investigate the modulation of expression of hBDs in cultured primary bronchial epithelial cells (PBEC) by rhinovirus-16 (RV16), a respiratory virus responsible for the common cold and associated with asthma exacerbations. RV16 was found to induce expression of hBD-2 and -3 mRNA in PBEC, but did not affect hBD-1 mRNA. Viral replication appeared essential for rhinovirus-induced beta-defensin mRNA expression, since UV-inactivated rhinovirus did not increase expression of hBD-2 and hBD-3 mRNA. Exposure to synthetic double-stranded RNA (dsRNA) molecule polyinosinic:polycytidylic acid had a similar effect as RV16 on mRNA expression of these peptides in PBEC. In line with this, PBEC were found to express TLR3, a Toll-like receptor involved in recognition of dsRNA. This study shows that rhinovirus infection of PBEC leads to increased hBD-2 and hBD-3 mRNA expression, which may play a role in both the uncomplicated common cold and in virus associated exacerbations of asthma. PMID- 12900057 TI - Various cells of the immune system and intestine differ in their capacity to reduce hexavalent chromium. AB - The cells of the immune system form a strong line of defence against foreign substances. The present study was undertaken to investigate the capacity of different cells of Wistar rats to reduce potentially carcinogenic hexavalent chromium (Cr-VI) into less toxic trivalent chromium in vitro. 5 x 10(6) cells were incubated with 10 or 25 microg ml(-1) of Cr (VI) in the form of K2Cr2O7 at 37 degrees C in the presence of 5% CO2 in air. At various time periods the remaining amount of Cr (VI) was measured and the percentage of Cr (VI) reduced was calculated. Among the single cell suspensions from the splenic cells a peak reduction of 55% was observed with the total spleen cells, 40% with the B lymphocyte-enriched subpopulation, 10% with T-lymphocytes and 24% with the macrophages. The reduction by splenic and peritoneal macrophages was similar. Total thymocytes reduced 54% of the Cr (VI). Since the most common route of entry of chromium is through drinking water and food, intestinal cells were also investigated. Among the intestinal cells the maximum reduction of 100% (of 10 microg ml(-1)) was observed with the upper villus cells and 72% with the middle villus cells while reduction was the least (4%) with the crypt cells. The reduction in the intestinal loop in situ was 100%. The time taken by each cell type for the peak reduction to Cr (VI) was markedly different. The findings thus show that the capacity of different cells in the body differs vastly in their capacity and time taken to reduce hexavalent chromium. The most efficient handling of Cr (VI) by the intestine, due to the presence of a variety of cells and bacteria, protects the body from its adverse effects. PMID- 12900059 TI - The acute phase protein serum amyloid A primes neutrophils. AB - We studied here the effect of the acute phase protein serum amyloid A (SAA) on the oxidative burst of neutrophils. Incubation of neutrophils with SAA increased the rate of oxygen uptake and the production of reactive oxygen species of neutrophils activated with opsonized zymosan (OZ). The increment in the neutrophil oxidative burst was dependent on SAA concentration in the range of 3 33 microg protein ml(-1) and was observed only in the presence of a relatively low amount of OZ (1 x 10(6) particles ml(-1)). SAA did not affect oxygen consumption and reactive oxygen production triggered by other stimuli, such as f Met-Leu-Phe, phorbol myristate acetate or non-opsonized zymosan. Our finding points to a priming effect of SAA probably associated with mobilization of receptors for opsonized particles and strengthens the role of SAA as an effector of neutrophil functions in inflammation. PMID- 12900058 TI - Osteoclast differentiation by human osteoblastic cell line SaOS-2 primed with bacterial lipid A. AB - We examined the responses of human osteoblastic cell line SaOS-2 to bacterial lipid A, a bioactive center of lipopolysaccharide, during osteoclast differentiation of human peripheral blood mononuclear cells (PBMC). SaOS-2 cells expressed mRNA for Toll-like receptor (TLR) 4, MD-2, CD14, and myeloid differentiation factor 88, whereas they failed to express mRNA for TLR2. Escherichia coli-type synthetic lipid A (compound 506) induced cytokine mRNA expression and nuclear factor (NF)-kappaB activation in SaOS-2 cells. Compound 506 also increased the expression of receptor activator of NF-kappaB ligand. Further, cells primed with compound 506 augmented the differentiation of PBMC into osteoclastic cells, and the effect was inhibited by anti-TLR4 monoclonal antibody. These findings suggest that the TLR signaling cascade in osteoblastic cells is involved in regulating the function of osteoclastogenesis. PMID- 12900060 TI - Retrospective detection of a subclinical hepatitis A virus (HAV) epidemic affecting juvenile cohorts of the Hungarian population. AB - Sero-epidemiological surveys of serum samples taken in 1982, 1987, 1994 and 1999 have been performed with hepatitis A virus-specific (HAV-specific) serological tests. Results obtained during these surveys show that the proportion of seropositive blood donors decreased from 69% to 18% within 17 years. The authors have recognised a (mainly subclinical) epidemic, affecting about 115000 teenagers in 1992-1994 in Hungary, is a threatening phenomenon. It was calculated that only about 3600 clinical diseases were associated with the epidemic, recognised retrospectively from the findings of the four sero-epidemiological surveys. Epidemiological data indicated that the excess clinical diseases caused by HAV concentrated in the southern counties of Hungary, which have been affected by the social and military activities between 1992 and 1994. Due to the decrease of subjects seropositive for HAV, sera from preselected or actively immunised donors will be required in the future and vaccination against HAV with killed virus is likely to be recommended for risk groups. Furthermore, health authorities might promote active immunisation of young children against HAV infection; for that, promotion of manufacturing combination vaccines of HAV/HBV/DPT or, for certain countries, HAV/DPT would be desirable. PMID- 12900061 TI - Continuing challenges for computer-based neuropsychological tests. AB - A number of issues critical to the development of computer-based neuropsychological testing systems that remain continuing challenges to their widespread use in occupational and environmental health are reviewed. Several computer-based neuropsychological testing systems have been developed over the last 20 years, and they have contributed substantially to the study of neurologic effects of a number of environmental exposures. However, many are no longer supported and do not run on contemporary personal computer operating systems. Issues that are continuing challenges for development of computer-based neuropsychological tests in environmental and occupational health are discussed: (1) some current technological trends that generally make test development more difficult; (2) lack of availability of usable speech recognition of the type required for computer-based testing systems; (3) implementing computer-based procedures and tasks that are improvements over, not just adaptations of, their manually-administered predecessors; (4) implementing tests of a wider range of memory functions than the limited range now available; (5) paying more attention to motivational influences that affect the reliability and validity of computer based measurements; and (6) increasing the usability of and audience for computer based systems. Partial solutions to some of these challenges are offered. The challenges posed by current technological trends are substantial and generally beyond the control of testing system developers. Widespread acceptance of the "tablet PC" and implementation of accurate small vocabulary, discrete, speaker independent speech recognition would enable revolutionary improvements to computer-based testing systems, particularly for testing memory functions not covered in existing systems. Dynamic, adaptive procedures, particularly ones based on item-response theory (IRT) and computerized-adaptive testing (CAT) methods, will be implemented in new tests that will be more efficient, reliable, and valid than existing test procedures. These additional developments, along with implementation of innovative reporting formats, are necessary for more widespread acceptance of the testing systems. PMID- 12900062 TI - Further standardization of some NES3 tests. AB - NES3 is a computer-based neurobehavioral testing system designed for use in investigating potential cognitive impairment. Data from NES3 tests employed in a study of epilepsy self-management were analyzed to estimate test-retest reliability for NES3 tests on a large sample and to estimate the effect of some common covariates of test performance. A total of 319 participants in an epilepsy self-management study were examined on three occasions (baseline, 3-month follow up, and 6-month follow-up) with a set of psychological questionnaires and seven neuropsychological tests from NES3. Test-retest correlations were calculated between measures obtained at baseline and at 3 months. Principal components analysis was performed on the baseline data. The potential effects of covariates (age, education, reading test scores, depression status, and examiner) were investigated in regression models. Three-month test-retest correlations were excellent for Adult Reading Test (ART) (r=0.95), strong for Digit-Symbol (r=0.82), Sequence B (r=0.79), and Sequence A (r=0.76); and modest (r's between 0.56 and 0.67) for Digit Span Forward and Backward, Visual Span Forward and Backward, and Pattern Memory. Alternate-forms correlations were strong for HVLT (r's between 0.71 and 0.82). Principal components analysis yielded four interpretable components. Age and reading score were significant covariates of virtually all of the test summary measures, while education, gender, race, and depression were not generally significant covariates. Changes to the method of calculation of some summary measures, changes to the initial instructions to the subjects, and addition of correctional feedback to subjects during the tests appeared to improve the reliability of some NES3 tests. Implementation of the HVLT and ART in computer-assisted format added breadth of coverage to the battery. NES3 tests may provide reliable, efficient data for use in epidemiologic studies of potential cognitive effects of occupational and environmental exposures. PMID- 12900063 TI - A computer-based reading test for use as an index of premorbid general intellectual level in North American English-speaking adults. AB - Brief tests of vocabulary or reading ability are often used as an index of general intellectual level to hypothesize deficits in patients, to establish comparability of comparison groups, and as a covariate of performance on other neuropsychological tests. We evaluated a new variant of a reading test for potential use in estimating premorbid general intellectual ability and implementation as a computerized-adaptive test. A total of 319 outpatients who participated in an epilepsy self-management study were tested on three occasions, completing the Wide-Range Achievement Test-3 (WRAT-3) Reading test at baseline and the NES3 Adult Reading Test (ART) at the 3-month follow-up. At the 6-month follow-up, some participants completed the ART and some the NES2 Vocabulary test. ART and WRAT-3 Reading scores obtained 3 months apart were highly correlated (r=0.87, n=179), while ART and NES2 Vocabulary scores were somewhat less correlated (r=0.80, n=61). The ART test-retest correlation was high (r=0.95, n=79). The ART distribution was spread out more evenly at the high end than that of the WRAT-3 Reading. ART score was significantly related to education, race, income, and examiner category, but was unrelated to age category and depression status. Non-significant gender differences were observed. An item-response theory (Rasch model) analysis showed good fit of the model to the observed data and yielded an improved ranking of the ART items in order of difficulty. The NES3 ART exhibited high reliability and strong association with WRAT-3 Reading. These results suggest that the NES3-ART may be useful as an index of general intellectual ability for comparison of groups and for use as a covariate in data analyses among English-speaking, North American adults. The Rasch model analysis provides a basis for implementing the ART as a computerized-adaptive test, which should improve its time efficiency and reduce participant burden in taking the test. PMID- 12900064 TI - Measures of short-term test-retest reliability of computerized neurobehavioral tests. AB - Neurobehavioral test batteries are often administered repeatedly to evaluate changes over time or effects of clinical interventions or neurotoxic exposures. Time intervals between test sessions range from very short (hours) to very long (decades). The aim of this study was to compare the impact of two brief time intervals on the test-retest reliability of frequently used neurobehavioral tests. Fifty healthy adults were recruited through newspaper advertisements in Portland, Oregon. Participants were divided into either a 6h (same-day) or 1-week retest group. All participants completed a battery of tests from the computerized Behavioral Assessment and Research System (BARS). Reliability was assessed by Pearson product-moment correlation and by intraclass correlation coefficient (ICC). The test battery generally showed adequate reliability in the short-term (week) and very short-term (day) and stability in performance over repeated administration when examined by multiple measures. Intraclass correlation coefficient ranged from 0.35 to 0.85. The magnitude of variation of performance in the administered tests was equally distributed around zero (i.e. no difference). The findings suggest that neurobehavioral tests such as BARS may be a useful tool for the assessment of acute exposures and clinical status where short-term evaluation is required (e.g. in the same-day or within 1 week). PMID- 12900066 TI - Cross-cultural comparison of neurobehavioral performance in Asian workers. AB - Widely-used neurobehavioral tests have been developed and standardized on Western populations, but studies on subject factors for Asian populations have been very limited. For the effective application and interpretation of neurobehavioral tests in Asian populations, an evaluation of the effects of subject factors, including cultural background, is necessary. A cross-cultural study was conducted to evaluate the effects of cultural background and the interaction between cultural background and education on neurobehavioral tests in Asian populations. The Korean version of the Swedish Performance Evaluation System (Simple Reaction Time, Symbol Digit, and Finger Tapping Speed) and a pegboard test were administered to 537 workers who were not exposed to chemicals at work from Fareast (Korea and Chinese), Central (Uzbekistan and Tajikistan), and South Asia (Sri Lanka and Indonesia). The Fareast Asian group exhibited better performance in adjusted test scores than other Asian groups, achieving significance for Symbol Digit and Finger Tapping Speed in both genders. The magnitude of the effect of cultural background on Symbol Digit was comparable to the effect of about 10 years of education. Cultural background did not modify the relation between years of education and Symbol Digit in either males or females. This study may provide the first evidence that cultural background has a large impact on neurobehavioral test performance, even within Asian populations, and suggests that cultural background is a critical confounding factor that must be controlled in epidemiologic studies which include Asian populations in the sample. PMID- 12900065 TI - Development of the Behavioral Assessment and Research System (BARS) to detect and characterize neurotoxicity in humans. AB - The Behavioral Assessment and Research System (BARS) is a computer-based testing system designed to assess neurobehavioral function in humans. It was developed to provide a series or battery of neurobehavioral tests optimized for the detection of neurotoxicity in non-mainstream human populations, specifically people with limited education or literacy. Key to meeting this goal were simply-stated instructions divided into an elemental series of steps, a 9BUTTON response unit to replace the computer keyboard for responding, and spoken instructions. Modifications all underwent serial testing in target populations to successively hone the changes to be more effective. A similar process was followed when developing adjustable parameters, test reliability assessments, and when implementing these tests with populations from different cultural groups and children. The principles and experiences that guided the development of BARS should inform the development of future testing systems to ensure that the new tests can be used with non-mainstream populations, which may be increasingly subject to neurotoxic exposures. PMID- 12900067 TI - Evaluation of the French version of EUROQUEST: a questionnaire for neurotoxic symptoms. AB - BACKGROUND: EUROQUEST was designed to explore various neuropsychiatric symptoms caused by neurotoxic agents in occupational epidemiological studies. This paper was aimed at assessing (i) the internal consistency of the French version of EUROQUEST, (ii) its sensitivity to age, educational level and exposure, (iii) and its convergent validity by examining relationships between declared symptoms and results from psychometric tests on the same mental functions. DESIGN: Data from four cross-sectional studies were selected: 180 workers exposed to toluene, mixed solvents or anaesthetic gas, and 588 non-exposed ones. The internal consistency of the questionnaire was assessed by computing Cronbach's alphas. Sensitivity to age, educational level and exposure were assessed with multiple linear regression models. Relationships between EUROQUEST scores and correspondent neuropsychological tests were assessed with Pearson's correlation coefficients. The relationships between EUROQUEST domain specific scores, age and educational level were evaluated for men and women from the Reference group. Most of the analyses were performed in men and women separately. RESULTS: The results of the Cronbach's alphas computed for each EUROQUEST domain, showed a reasonable internal consistency for the questionnaire. For both women and men, memory problems and sleep disorders increased significantly with age. A significant association between EUROQUEST scores and exposure to neurotoxic agents was found for workers exposed to anaesthetic gas only. In addition, EUROQUEST memory symptoms from the Reference population were found to be correlated with two memory neuropsychological tests, both for men and women. CONCLUSION: Our results showed a high internal consistency and a good convergent validity on assessed domains. Moreover, they showed a significant sensitivity to age, educational level and highlighted sensitivity to anaesthetic gas exposure. PMID- 12900068 TI - Suboptimal performance on neuropsychological tests in patients with suspected chronic toxic encephalopathy. AB - Suboptimal performance during neuropsychological testing can seriously complicate assessment in behavioral neurotoxicology. We present data on the prevalence of suboptimal performance in a group of Dutch patients with suspected chronic toxic encephalopathy (CTE) after long-term occupational exposure to solvents. One hundred and forty-five subjects referred to one of two Dutch national assessment centers for CTE were administered the Amsterdam Short-Term Memory Test (ASTM) and the Test of Memory Malingering (TOMM), two tests specifically developed for the detection of suboptimal performance. For both tests, very cautious cut-off scores were chosen with a specificity of 99%. Results indicated that suboptimal performance appears to be a substantial problem in this group of patients with suspected CTE after long-term exposure to organic solvents. Only 54% of our subjects obtained normal scores on both tests of malingering, i.e. at or above cut-off score. The two tests seemed to measure the same concept in that nearly all the subjects with low TOMM scores also had low ASTM scores. However, a higher proportion of subjects scored below the cut-off on the ASTM than on the TOMM. PMID- 12900069 TI - Neuropsychological effects of ethylene dichloride exposure. AB - Production of 1,2-dichloroethane (EDC) has increased and exposure to it has been reported to produce adverse effects in animals and CNS effects in humans. A group of 221 EDC exposed hazardous clean-up workers incurred chronic exposure to EDC without protective equipment. A clinical history and the following tests were administered: the WHO Adult Environmental Neurobehavioral Test Battery (AENTB), Wechsler Adult Intelligence Scale (WAIS-III), Wechsler Memory Scale (WMS-III); WRAT 3 Reading, Cancellation H, Trail Making, Stroop, Rey Osterreith, Animal Naming, COWAT, TOMM, Rey 15, Lanthony d-15 Color Vision, and Vistech Contrast Sensitivity. Mood and symptoms were assessed with the SCL90-R, BDI, BAI and IES. The clinical neuropsychological evaluation of these workers indicates lower neuropsychological functioning in the domains of processing speed; attention; cognitive flexibility; motor coordination and speed; verbal memory; verbal fluency; and visuo-spatial abilities. These workers also showed disturbed mood and impaired vision. Dose/response relationships between exposure to EDC and test scores are shown. PMID- 12900070 TI - Inhalation of 1-bromopropane causes excitation in the central nervous system of male F344 rats. AB - The present study investigates the effects of 1-bromopropane (1BP) on animal behavior to determine the extent of toxicity to the central nervous system (CNS). We measured the spontaneous locomotor activity (SLA) of rats before and after 3 weeks of exposure to 1BP for 8 h per day. In control and 10 ppm groups, the SLA values were similar to pre-exposure levels on post-exposure Day 1 and thereafter. However, the SLA values in the 50 and 200 ppm groups were higher than pre exposure levels. Open-field behavior was evaluated after exposure and freezing time decreased with exposure to increasing concentrations of 1BP. Ambulation and rearing scores in the exposed groups were higher than control values, particularly in the 50 and 200 ppm groups. The frequency of defecation and urination decreased almost dose-dependently. Exposure to 50-1000 ppm of 1BP did not affect passive avoidance behavior examined using a step-through type apparatus. The amount of time swimming in the water maze test was not affected in the controls, or groups exposed to 50 and 200 ppm 1BP, but that in the 1000 ppm group was increased compared with control. Exposure at 50-1000 ppm dose dependently decreased the traction performance of rats, indicating decreased muscle strength. We found that 10-200 ppm of 1BP exposure did not affect motor coordination determined by rota-rod performance. The increased SLA values and open-field activity support the notion that 1BP has excitatory effects on the CNS of F344 male rats. In addition, 1BP reduced the grip or muscle strength of the rats. Memory function was not disordered and the motor coordination of all four limbs remained normal. PMID- 12900071 TI - Neurobehavioral effects of pesticides: state of the art. AB - The authors have reviewed the literature on neurobehavioral toxicity of pesticides to assess the status of knowledge on this matter. Some data suggest that exposure to DDT and fumigants may be associated with permanent decline in neurobehavioral functioning and increase in psychiatric symptoms, but, due to the limited number of studies available and the scarce knowledge on exposure levels, no firm conclusion can be drawn. Data on subjects acutely poisoned with organophosphorous compounds suggest that an impairment in neurobehavioral performance and, in some cases, emotional status may be observed as a long-term sequela, but the possibility still remains that these effects were only an aspecific expression of damage and not of direct neurotoxicity. Studies carried out on subjects chronically exposed to organophosphates, but never acutely poisoned, do not provide univocal results but the slight changes consistently observed in sheep dippers suggest the need of focusing on activities characterized by relatively higher exposure levels. In general, the main limits of existing knowledge are the variability of the testing methods used, which makes it difficult to compare the results of single studies, and the scarce knowledge on exposure levels. A promising approach may be the conduction of prospective longitudinal or cohort studies, where exposure and dose assessment can be more easily controlled, or the evaluation of cohorts of workers a priori selected for the availability of environmental and biological monitoring data. The follow up of the populations under study may give an answer at the problem of the prognostic significance of the observed changes. Also the protocols used to assess neurobehavioral functioning need to be standardized. PMID- 12900072 TI - Neural injury biomarkers of novel shellfish toxins, spirolides: a pilot study using immunochemical and transcriptional analysis. AB - In 1991, routine biotoxin monitoring of bivalve molluscs at aquaculture sites along the eastern shore of Nova Scotia, Canada revealed a group of novel seafood toxins called spirolides, whose origin was the dinoflagellate Alexandrium ostenfeldii. Result from this preliminary study in rodents demonstrates a highly toxic lethal response in rats and mice after intraperitoneal injections of lipophilic extracts. To elucidate the modes of action and toxicologic pathology, brain and internal organs were examined by histology and various biomarkers of neural injury were monitored by immunohistochemistry (IH) and/or transcriptional analysis. The histological and transcriptional data showed that the effects of spirolides are species dependent for mice and rats. Histopathology showed that in the mouse brain, the hippocampus and brain stem appeared to be the major target regions but no histological changes were observed in the rat. Transcriptional analysis in the mouse brain showed no alterations in the biomarkers whereas in the rat brain there were major changes in the markers of neuronal injury. These biomarkers included the early injury markers HSP-72, c-jun and c-fos which are essential for converting stimuli into intracellular changes within neurons. The potential effects of spirolides were also evaluated with respect to different subtypes of the acetylcholine receptors (AChRs) since earlier reports showed these as putative targets. Both the muscarinic and nicotinic AChRs were found to be upregulated. Hence, transcriptional and immunohistochemical analysis does provide insight to the molecular mechanisms of this novel group of shellfish toxins. No histological changes were observed in other tissues. PMID- 12900073 TI - Application of a latent variable model for a multicenter study on early effects due to mercury exposure. AB - A latent variable model was applied to the results of an Italian multicenter nation-wide cross-sectional study to assess the earliest health effects due to mercury (Hg) exposure caused by occupation, dental amalgams (DENTAM) and fish eating (FISH). The studied population included subjects recruited from four different geographical areas. A total number of 122 workers from chloroalkali plants and production of thermometers and neon lamps formed the occupationally exposed group, whereas 196 subjects, recruited from the same areas and not occupationally exposed to mercury, formed the control group. Neuropsychological functions were assessed with neurobehavioral testing including vigilance, motor and cognitive function, tremor measurements, and with symptoms concerning neuropsychological and mood assessment. Neuroendocrine function was examined with the measurement of prolactin (PRL) level. Parameters of immunological and renal function were also measured. The target population was characterized by the number and surface of dental amalgams and consumption of fish. In the exposed workers the average urinary mercury (U-Hg) was 10.4+/-6.9 (geometric mean 8.3, range 0.2-35.2) microg/g creatinine, whereas in the control group it was 1.9+/ 2.8 (geometric mean 1.2, range 0.1-33.2) microg/g creatinine. The preliminary results indicated that finger tapping (FT) and the Branches alternate movement task (BAMT) coordination test were associated with the occupational exposure (OCCEXP). PRL was significantly decreased among the exposed workers, and inversely related to U-Hg. Among the immunological and renal parameters, cytokine serum interleuchin-8 (sIL8) and beta(2) micro globulin (beta(2)MG) were lower in the exposed group and negatively correlated to U-Hg. Small-size fish consumption was associated to a beneficial effect on symptoms reporting. No effects were observed concerning dental amalgams. After first evaluating the relationship between mercury exposure and each indicator of effect, further assessment was performed to identify the earliest effects related to mercury exposure among those who resulted in being associated in the preliminary elaboration. Two latent variables "exposure" and "effect" were identified, integrating respectively the different forms of exposure (occupational, due to dental amalgams and fish consumption) and the indicators of effects (FT, BAMT, PRL, sIL8, beta(2)MG). Confounding factors (age, alcohol, body mass index (BMI)) were considered in the same model. This further analysis showed that an inverse association of occupational exposure to mercury with PRL and BAMT, with Hg-U mediating the effect on PRL, was predominant with respect to the other form of mercury exposure, the other indicators of effect and the confounders. In conclusion, this study supports the finding of alterations of neuroendocrine secretion and motor coordination at very low occupational exposure levels of inorganic mercury, below the current ACGIH Biological Exposure Index. These changes occur at lower levels than other subtle effects on the renal function and the immunitary system. On the contrary, dental amalgams and small-size fish consumption do not seem to be associated to any adverse health effect at these exposure levels. PMID- 12900074 TI - Sub-clinical neurobehavioral abnormalities associated with low level of mercury exposure through fish consumption. AB - In order to assess early neurotoxic effects associated with relatively low levels of mercury absorbed through fish eating, two groups of 22 adult male subjects, habitual consumers of tuna fish, and 22 controls were examined using a cross sectional field study. The assessment included neurobehavioral tests of vigilance and psychomotor function, hand tremor measurements and serum prolactin assessment. Mercury in urine (U-Hg) and serum prolactin (sPRL) were measured in all exposed subjects and controls, whereas measurements of the organic component of mercury in blood (O-Hg) were available for only 10 exposed and six controls. U Hg was significant higher among exposed subjects (median 6.5 microg/g of creatinine, range 1.8-21.5) than controls (median 1.5 microg/g of creatinine, range 0.5-5.3). The median values of O-Hg were 41.5 microg/l among the tuna fish eaters and 2.6 microg/l in the control group. Both U-Hg and O-Hg were significantly correlated with the quantity of fish consumed per week. Significant differences in sPRL were found between exposed (12.6 ng/ml) and controls (9.1 ng/ml). Individual sPRL were significantly correlated with both U-Hg and O-Hg levels. The neurobehavioral performance of subjects who consumed tuna fish regularly was significantly worse on color word reaction time, digit symbol reaction time and finger tapping speed (FT). After considering the education level and other covariates, the multiple stepwise regression analysis indicated that O-Hg concentration was most significantly associated with individual performance on these tests, accounting for about 65% of the variance in test scores. PMID- 12900075 TI - Association of chronic and current measures of lead exposure with different components of brainstem auditory evoked potentials. AB - Current blood lead (PbB) affects brainstem auditory evoked potentials (BAEPs) in children but whether a similar association exists in lead-exposed adults remains unclear. During an investigation of the neurobehavioral effects of occupational lead exposure we performed BAEPs on 359 English- and French-speaking, currently exposed, male, lead smelter workers having a mean (S.D.) age of 41 (9.0) years, employment duration of 17 (7.9) years, PbB of 28 (8.4)microg/dl, working-lifetime weighted average blood lead (TWA) of 39 (11.9) microg/dl, and working-lifetime integrated blood lead (IBL) index of 719 (421.0) microg.year/dl, the latter a measure of cumulative lead dose. BAEPs were performed at a click stimulation of 10s(-1) for 1000 repetitions, at an intensity 75dB above the threshold of the ear tested. Right-sided latencies for peak waves I, III, and V and the corresponding interpeak intervals (IPI) I-V, I-III, and III-V were chosen for analyses. Age correlated significantly with BAEPs, PbB, TWA, and IBL. Partial correlation analyses adjusting for age found PbB and TWA significantly associated with wave I latency r=0.13, P<0.01 and r=0.11, P<0.05, respectively, and IBL significantly associated with wave III latency r=0.16, P<0.01. The contribution of age, PbB, TWA and IBL to the variances of different BAEPs was assessed using multiple regression analysis. In the regression model of the full group, after the contribution of age, PbB and TWA accounted for significant variance of wave I, Deltar(2)=1.8, P<0.01 and Deltar(2)=1.2%, P<0.04, respectively, and IBL accounted for significant variance of wave III latency, Deltar(2)=2.8%, P=0.00 and I-III interpeak interval, Deltar(2)=1.4%, P<0.03. Four groups similar in age were created with increasing abnormalities based upon clinical cut-off scores for wave I latency and I-V interpeak interval. PbB, TWA, and IBL were significantly higher in the group with abnormalities of both latency in wave I and IPL I-V. Lead exposure interferes with BAEPs in a dose-dependent manner. Current lead exposure in this population of lead smelter workers preferentially affected conduction in the distal auditory nerve while chronic lead exposure appeared to impair conduction in the auditory nerve and the auditory pathway in the lower brainstem. PMID- 12900076 TI - Manganese intoxication: the cause of an inexplicable epileptic syndrome in a 3 year old child. AB - Excess manganese (Mn) can cause several neurotoxic effects, however only a few studies have reported epileptic syndromes related to manganese intoxication. We describe an epileptic syndrome due to manganese intoxication in a 3 year old male child. His blood manganese was elevated, but no other abnormal values or toxic substances were found in blood or urine. The electroencephalogram (EEG) showed a picture of progressive encephalopathy, while brain magnetic resonance was normal. The patient's conditions rapidly worsened to epileptic status despite the use of antiepileptic drugs. Chelating treatment with CaNa(2)EDTA was initiated to remove excess manganese and promptly succeeded in reverting epileptic symptoms. Concurrently, manganese blood levels and electroencephalogram progressively normalized. Thereafter it has been possible to discontinue antiepileptic treatment, and the patient remains in excellent conditions without any treatment. PMID- 12900077 TI - Blood manganese and alcohol consumption interact on mood states among manganese alloy production workers. AB - Long-term exposure to manganese (Mn) can induce neurotoxic effects including neuromotor, neurocognitive and neuropsychiatric effects, but there is a great interpersonal variability in the occurrence of these effects. It has recently been suggested that blood Mn (MnB) may interact with alcohol use disorders, accentuating neuropsychiatric symptoms. The objective of the present study was to explore a possible interaction between alcohol consumption and MnB on mood states, using an existing data set on Mn exposed workers. Respirable Mn exposure in the plant averaged 0.23mg/m(3) and was correlated with MnB. All participants for whom all data on MnB concentration and mood (assessed with the Profile of Mood States (POMS)) were available and who reported currently drinking alcohol were included in the analyses (n=74). Workers were grouped according to their MnB concentration (<10 and > or =10 microg/l) and alcohol consumption (<400 and > or =400g per week). Two-way ANOVAs were performed on each POMS scale and Mann Whitney tests were used to assess group differences. Workers in the higher alcohol consumption group had higher scores on three POMS scales: tension, anger and fatigue. There was no difference for POMS scale scores between MnB subgroups. Dividing the group with respect to alcohol consumption and MnB showed that the group with high alcohol consumption and high MnB displayed the highest scores. In the lower MnB category, those in the higher alcohol consumption group did not have higher scores than the others. The interaction term for alcohol consumption and MnB concentration was statistically significant (P<0.05) for the depression, anger, fatigue and confusion POMS scales. There was a tendency for tension (P<0.06), and it was not significant for vigor. This study shows the first evidence of an interaction between MnB and alcohol consumption on mood states among Mn exposed workers and supports the results from a previous population based study. PMID- 12900078 TI - Nervous system effects of occupational manganese exposure on South African manganese mineworkers. AB - Occupational exposure to airborne manganese dust has been shown to produce adverse effects on the central nervous system. Four hundred and eighty-nine blue and white collar manganese mineworkers from South Africa were studied cross sectionally to investigate the nervous system effects of medium to low occupational manganese exposures. The different facilities included underground mines, surface processing plants, and office locations. A job exposure matrix was constructed using routine occupational hygiene data. Exposure variables included years of service, a cumulative exposure index (CEI) and average intensity of exposure (AINT) across all jobs, and blood manganese. Endpoints included items from the Q16, WHO-NCTB, SPES, and Luria-Nebraska test batteries, and a brief clinical examination. Potential confounders and effect modifiers included age, level of education, past medical history including previous head injury, previous neurotoxic job exposures, tobacco use, alcohol use and home language. Associations were evaluated by multiple linear and logistic regression modeling. Average exposure intensity across all jobs was 0.21mg/m(3) manganese dust. Multivariate analyses showed that none of the symptom nor test results were associated with any measure of exposure including blood manganese, after adjustment for confounders. This relatively large null study indicates that manganese miners exposed on average across all jobs to MnO(2) at levels near the American Conference of Governmental Industrial Hygienists Threshold Limit Value (ACGIH TLV) are unlikely to have a subclinical neurotoxicity problem. PMID- 12900080 TI - Manganese, monoamine metabolite levels at birth, and child psychomotor development. AB - Several studies have demonstrated neurobehavioral impairment related to manganese (Mn) exposure in the workplace. Exposure to high doses of manganese is associated with irreversible neurodegenerative disorders resembling idiopathic Parkinson disease. Although there is a risk of Mn accumulation in the foetus during pregnancy, little information exists about developmental effects of environmental low-level exposure in human. For this reason, we conducted a prospective epidemiological study in 247 healthy pregnant women and their babies to determine the long-term effect of in utero Mn levels on child's psychomotor development. Concurrently, we examined the relationship between Mn tissue levels at delivery and foetal plasma monoamine metabolites. Of the newborns, 195 were examined at 9 months, 126 at 3 years and 100 at 6 years. At 9 months, the Brunet-Lezine scales were administered. The McCarthy scales of children's abilities were used at 3 and 6 years. After adjustment for potential confounding co-factors (child's gender, mother's educational level), negative relationships were observed between cord blood Mn levels and several psychomotor sub-scales at age of 3 years: "attention" (partial r=-0.33, P<0.001), "non-verbal memory" (partial r=-0.28, P<0.01), and "hand skills" (partial r=-0.22, P<0.05). No significant relationships were observed between Mn measures at birth and the general psychomotor indices, Brunet Lezine developmental quotient (DQ) at 9 months or McCarthy general cognitive index (GCI) at 3 and 6 years. Maternal blood Mn levels were negatively associated with foetal plasma HVA and 5-HIAA concentrations (adjusted for labour duration, child's gender, and smoking during pregnancy), but the adjustment for monoamine levels at birth did not change the association between the Mn levels and the psychomotor scores. These results suggest that environmental Mn exposure in utero could affect early psychomotor development. PMID- 12900079 TI - Effect of manganese exposure on MPTP neurotoxicities. AB - We used a 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-treated mice model to evaluate whether manganese (Mn) exposure can affect MPTP-induced neurotoxicity. We randomly assigned adult male C57BL/6 mice (n=5-7 per group) the following treatments: SO, Mn(-) x MPTP(-); MO, Mn(+) x MPTP(-); SM, Mn(-) x MPTP(+); MM, Mn(+) x MPTP(+). Mn (MnCl(2).4H(2)O) was administered intraperitoneally at a dose of 2 mg/kg daily for 3 weeks. MPTP was then administered intraperitoneally at a dose of 30 mg/kg daily for 5 days in the SM and MM groups. Seven days after the last MPTP injection, the animals were sacrificed. Blood Mn levels were elevated in the Mn-exposed groups. Striatal Mn levels were not influenced by Mn treatment alone, however, they were decreased following MPTP. Tyrosine hydroxylase (TH)-immunoreactive (ir) neurons in the substantia nigra pars compacta (SNpc) were decreased significantly in the MPTP exposed groups. Densities of TH- and dopamine transporter (DAT)-ir axon terminals in the caudate-putamen (CPU) were also decreased in the MPTP-treated groups. Furthermore, glial fibrillary acidic protein (GFAP)-ir astrocytes increased in the CPU with MPTP treatment. However, no effects were observed with Mn exposure. Concentrations of dopamine (DA), 3,4-dihydrophenyl acetic acid (DOPAC) and homovanillic acid (HVA) in the corpus striatum were also decreased significantly with MPTP treatment alone, but Mn had no effect. Thus, decreased dopaminergic activities with MPTP led to decreased DA and its metabolites. Significant hypertrophies of GFAP-ir astrocytes in the globus pallidus (GP) were observed in Mn-exposed groups, especially in the MM group. MPTP targeted dopaminergic systems whereas Mn neurotoxicities occurred in the GP. In conclusion, our data suggest that Mn does not potentiate the neurotoxicity of MPTP. PMID- 12900081 TI - Occupational exposure to chemicals and sensory organs: a neglected research field. AB - The effect of industrial chemicals on the sensory perception of exposed workers has received scant attention from the medical community to date, and the scientific literature is mainly limited to some case-reports or isolated studies. Possible explanations for this include the complexity of sensory perception, and the lack of agreement among researchers on methods for testing large groups of subjects. Nevertheless, some published studies showed that vision, hearing and olfactory function can be affected by various industrial metals and solvents, and some data exist also for touch and taste. This review discusses the main industrial chemicals involved. The pathogenesis of the toxicity of chemicals to sensory perception may be related to an action on receptors, nerve fibers, and/or the brain; probably, different pathogenetic mechanisms are involved. One of the main problems in this research field is that most of the studies to date evaluated the effect of a single industrial chemical on a single sense: as an example, we know that styrene exposure can impair smell and also hearing and vision but we have little idea whether different senses are impaired in the same worker, or whether each impairment is independent. In addition, workers are frequently exposed to different chemicals: co-exposure may have no effect, or result in both an increase or a decrease of the effect, as was observed for hearing loss, but studies on this aspect are largely insufficient. Research shows that both occupational and environmental exposure to industrial chemicals can affect sense organs, and suggests that the decline of perception with age may be, at least partly, related to this exposure. Nevertheless, available evidence is incomplete, and is largely inadequate for an estimation of a "safe" threshold of exposure. Good quality further research in this field is needed. This is certainly complex and demands adequate resources, but is justified by the ultimate result: the possibility to prevent an avoidable part of the decline in sensory function with age. PMID- 12900082 TI - Color vision impairment in workers exposed to neurotoxic chemicals. AB - Recent research shows that occupational exposure to several solvents, metals and other industrial chemicals can impair color vision in exposed workers. Occupation related color vision impairment usually results in blue-yellow color discrimination loss or, less frequently, a combination of blue-yellow and red green loss. The eyes may be unequally involved, and the course is variable depending on exposure and other factors. The pathogenesis of occupational color vision loss has not been elucidated; it may be due to, e.g. a direct action of neurotoxins on receptors, possibly on the cone's membrane metabolism, and/or to an interference with neurotransmitters within the retina. Other possible pathogenetic mechanisms, such as a direct effect to the optic nerve, have also been suggested. Occupational color vision loss is usually sub-clinical, and workers are unaware of any deficit. It can be assessed using sensitive tests, such as the Farnsworth-Munsell 100 Hue (FM-100) or the Lanthony D-15 desaturated panel (D-15 d). The latter is the most widely used for studies in groups of exposed workers, and offers the possibility of a quantitative evaluation of the results by calculation of the Bowman's Color Confusion Index (CCI), or of the Vingrys' and King Smith's Confusion Index (CI). Other advantages of D-15 d are the possibility to perform the test directly at the workplace, and the reproducibility when performed in standardized conditions. In most cases, occupation-related color vision impairment is correlated to exposure levels, and has often been observed in workers exposed to environmental concentrations below the current occupational limit proposed by the ACGIH. Progression with increasing cumulative exposure has been reported, while reversibility is still discussed. Acquired color vision impairment related to occupational exposure to styrene, perchloroethylene (PCE), toluene, carbon disulfide, n-hexane, solvent mixtures, mercury and some other chemicals are discussed. Results show that color vision testing should be included in the evaluation of early neurotoxicity of chemicals in exposed workers. The D-15 d would be useful in the surveillance of workers exposed to solvents and other chemicals toxic to the visual system. PMID- 12900083 TI - Visual dysfunction in workers exposed to a mixture of organic solvents. AB - The aim of this study was to investigate the relationship between the workers occupationally exposed to a mixture of organic solvents and their visual functions. Here the visual functions included color vision (CV), visual contrast sensitivity (CS) and visual evoked potentials (VEP). Test subjects were 182 workers at 53 furniture factories in the same industrial area of Japan. As control, a group consisted of 96 workers without exposure to any organic solvent was also tested. Exposure assessments were made both by the environmental concentration and biological monitoring. CV and CS tests were carried out for all the subjects. VEP was measured for 21 exposed subjects who were considered to have impaired CV and CS. In the results, the color confusion index (CCI) values of the exposed subjects were significantly higher than that of the age-matched controls (P<0.01). Their CS values were significantly lower than those in the controls at spatial frequencies of 6 and 12 cycles per degree (cpd) (P<0.01 and <0.05, respectively). A significant correlation between the concentration of urinary methylhippuric acid and contrast sensitivity was found by a multiple regression analysis (P<0.05). CCI showed a negative correlation at all spatial frequencies of CS in a simple regression analysis, no abnormal data were found by the VEP test in the exposed subjects who were found to have impaired CV and CS. The results suppose that a low concentration of the mixed organic solvents might affect the retina and optic nerve. However, it needs to be further researched if such an impact affects the Brodmann's areas of visual cortex in the brain. PMID- 12900084 TI - Color discrimination impairment in workers exposed to mercury vapor. AB - OBJECTIVE: To study color discrimination impairment in workers exposed to elemental mercury (Hg) vapor. SUBJECTS: Twenty-four male workers from a chloralkali plant exposed to Hg vapor, aged 42+/-9.8 years, duration of exposure 14.7+/-9.7 years, were examined. The 8h TWA air-borne Hg concentration in workplace was 59 microg/m(3); mean Hg urinary excretion (HgU) was 20.5+/-19.3 microg/g creatinine; mean Hg urinary excretion after the administration of a chelating agent, sodium 2,3-dimercapto-1-propane-sulfonate (DMPS), was 751.9+/ 648 microg/48h. Twenty-four age- and gender-matched control subjects were compared. Visual acuity, alcohol intake, smoking habits, and history of diseases or drugs potentially influencing color vision were registered. METHODS: The Lanthony 15-Hue desaturated test (L-D15-d) was used to assess color vision. The results were expressed quantitatively as Bowman's Color Confusion Index (CCI), and qualitatively according to Verriest's classification of acquired dyschromatopsias. RESULTS: The CCI was significantly higher in the exposed group than in the control (mean CCI 1.15 versus 1.04; P=0.04). The proportion of subjects with errorless performance on the Lanthony test was significantly lower in the Hg exposed group compared to referents (52% versus 73%; P=0.035). The exposed group showed higher frequency of type III dyschromatopsias (blue-yellow confusion axis) in comparison with the control group (12.5% versus 8.3%), however, the difference did not reach statistical significance. Multiple regression did not show any significant relationship between the CCI, and age, alcohol consumption, or measures of exposure. CONCLUSION: In agreement with previous studies by Cavalleri et al. [Toxicol. Lett. 77 (1995) 351; Environ. Res. Sec. A 77 (1998) 173], the results of this study support the hypothesis that exposure to mercury vapor can induce sub-clinical color vision impairment. This effect was observed at an exposure level below the current biological limit for occupational exposure to mercury. This raises doubts on the actual protection afforded by this limit concerning the effect of mercury on color vision. PMID- 12900085 TI - Olfactory function in workers exposed to moderate airborne cadmium levels. AB - OBJECTIVE: The present study investigated the olfactory function in workers exposed to moderate airborne cadmium (Cd) levels, with the purpose of identifying possible early adverse effects of the metal, not demonstrable with the traditional diagnostic methods. METHODS: The exposed group consisted of 33 men employed in cadmium fusion, sintering and alloys lamination. Two reference groups were considered: the first consisted of 39 subjects assigned to manual workings, but not exposed to harmful substances for olfaction (drivers, warehousemen); the second was characterized by 23 subjects exposed to iron and steel welding fumes (iron base alloys), non-specific irritants on the respiratory tract. Olfactory threshold and odor identification ability were separately quantified. Individual occupational exposure was studied by calculating the mean blood and urinary cadmium values (CdB and CdU, respectively) of the five years period during which, for each worker, the highest levels of dose indicators were measured. As indicator of renal tubular damage, urinary beta(2)-microglobulin levels in the same quinquennium were monitored. RESULTS: Mean olfactory threshold scored significantly higher in Cd workers (-5.26 log(10)v/v, P=0.02) than did in controls (welders: -5.78 log(10)v/v; P=0.26 compared to non-exposed -6.37 log(10)v/v). The odor identification test findings for Cd workers were similar to those of the reference groups. Moderate blood and urine cadmium levels (mean CdB 3.7 microg/l; mean CdU 4.4 microg/g creatinine) accompanied threshold impairment. Olfactory threshold weakening was confirmed in the subgroup of Cd workers in which urinary beta(2)-microglobulin never exceeded 300 microg/l (-5.41 log(10)v/v; P=0.045 compared to non-exposed). CONCLUSIONS: These data suggested that possible early toxic effects of the metal can occur at low levels, close to the limits proposed by the American Conference of Governmental Industrial Hygienists (ACGIH), confirming the hypothesis that primary olfactory neuron may represent the early target for cadmium toxic action. The action of the metal seemed to be due to an elective tropism for the olfactory epithelium and not to a non-specific irritant effect on the nasal cavity. The mechanism of this effect did not appear linearly dose-related. These findings underline the importance of olfactory tests to identify the early effects of xenobiotics even at low-exposure levels, and to contribute to verify the adequacy of the current exposure limits. PMID- 12900086 TI - Assessment of visual functions following prenatal exposure to organic solvents. AB - Prenatal exposure to organic solvents has been previously associated with increased risk of color vision deficits and reduced visual acuity in young children. These findings prompted us to evaluate visual functioning in solvent exposed infants using more sensitive non-invasive visual evoked potential (VEP) techniques. VEP techniques are described in the context of an ongoing prospective longitudinal cohort study of infants exposed to organic solvents in utero. VEPs are recorded via three active electrodes fitted over the occipital cortex while infants view changing visual stimuli. The sweep VEP is used to assess contrast detection and visual acuity by presenting sinusoidal gratings that "sweep" across a range of contrasts and spatial frequencies. Transient VEPs are used to assess responses to equiluminant chromatic- and luminance-modulated sinusoidal gratings presented in pattern onset-offset format. A single case study is presented showing abnormal chromatic responses and reduced contrast sensitivity in a 2.5 year-old boy following prenatal exposure to perchloroethylene (PCE). These VEP techniques therefore appear promising for the clinical assessment of visual toxicity in pediatric populations. PMID- 12900087 TI - Evidence of neurotoxicity in working children in Lebanon. AB - The potential for exposure to neurotoxicants in Lebanon is high, especially in the absence of strict control and regulatory measures in workplaces. Children working in mechanical and other trade workshops are at significant risk of exposure to organic solvents and as a result at significant risk to develop clinical and subclinical signs of neurotoxicity. This study investigated the association between exposure to solvents and neurobehavioral performance of 10-17 years old working children in comparison to groups of non-exposed working children and non-exposed children at school. A convenience cross-sectional sample of 100 working children (10-17 years) exposed to organic solvents at work, a positive control of 100 non-solvent exposed working children, and a negative control of 100 non-working, non-exposed school children were recruited to the study. The exposed and non-exposed groups were, as far as possible, frequency matched on geographic location of residence and on age. Neurotoxic effects were assessed through a questionnaire and the child's performance on a battery of neurobehavioral tests, which were specifically selected for a non-English speaking population and were tailored to assess the specific modalities of the central nervous system commonly known to be affected by the neurotoxicants identified. Children in the non-exposed groups (working and non-working) performed better than the working exposed group on reaction speed for the choice reaction time, symbol-digit and dual task tracking tests, but accuracy of performance was not affected on these tests. Children in the working exposed group also complained of more headaches, loss of concentration, memory deficits and higher irritability. This study identifies an association between exposure to solvents and lower neurobehavioral performance. Response speed appears to be particularly affected in solvent exposed working children. PMID- 12900088 TI - Developmental aspects of blood-brain barrier (BBB) and rat brain endothelial (RBE4) cells as in vitro model for studies on chlorpyrifos transport. AB - The mammalian central nervous system (CNS) is characterized by the blood-brain barrier (BBB), a restrictive barrier endowed with the maintenance of homeostatic control of an optimal milieu within the brain. Whereas in tissues other than the CNS, concentrations of various metabolites (amino acids, K+) can undergo frequent fluctuations, the CNS must keep rigorous control over the extracellular cerebral fluid composition, preventing the mirroring of transient fluctuations in blood, because abrupt changes in these metabolites can translate to aberrant CNS function. The BBB is a specialized structure accomplished by individual endothelial cells that are continuously linked by tight junctions. This brief review will address pertinent issues to development of the BBB. Particular emphasis will be directed at the role of astrocytes in the induction and maintenance of the restrictive properties of this barrier, and the utility of in vitro culture models in surveying transport kinetics, exemplified by recent studies with the pesticide, chlorpyrifos. PMID- 12900089 TI - The effects of chronic arsenic exposure from drinking water on the neurobehavioral development in adolescence. AB - This cross-sectional study examined the possible influence on the development of cognitive function among adolescents due to long-term arsenic exposure. Forty nine junior school students drinking arsenic-containing well water and 60 controls matched with age, sex, education, body height, body weight, body mass index, and socioeconomic status were compared. The former was divided into two groups: high and low exposure, with mean cumulative arsenic levels of 520629.0+/ 605824.2 and 13782.2+/-12886.0 ppm, respectively. Four neurobehavioral tests including continuous performance test (CPT), symbol digit (SD), pattern memory (PM) and switching attention (SA) were applied. A strong correlation between age and education caused collinearity in the multiple regression model (r=0.84, P<0.0001). Only education and sex, excluding age, were entered into the model as covariates. Pattern memory and switching attention were significantly affected by long-term cumulative exposure to arsenic after adjusting for education and sex. It is suggested that the arsenic levels in the well water may be monitored extensively, but if there is no intervention, then neurobehavioral function will not be protected. Limitations of the current study require replication of this effect in other studies to confirm this conclusion. PMID- 12900090 TI - From patient education to communication in health care. PMID- 12900091 TI - Shared decision making. PMID- 12900092 TI - Patient perceptions that limit a community-based intervention to promote participation. AB - A workshop designed to teach seniors to communicate more effectively with their physicians and enhance patient participation in the consultation was held in a community centre. A grounded theory analysis of follow-up telephone interviews provided examples of effectiveness but also revealed six categories of barriers to changing the pattern of established communication, particularly over the short term. PMID- 12900093 TI - Validation of a tool to assess health practitioners' decision support and communication skills. AB - As patients become more involved in decisions affecting their health, it is important to monitor and improve the support clinicians provide to facilitate shared decision making. The Decision Support Analysis Tool (DSAT) was developed as a research tool to evaluate practitioners' use of decision support and related communication skills during a clinical encounter. The DSAT, consisting of six categories of decision support skills and four categories of communication skills, was tested with 34 actual transcripts of patient-physician dialogue. The patients were prepared for the clinical encounter with either a detailed decision aid plus worksheet (n=16) or a pamphlet (n=18). Pairs of raters, blinded to the intervention allocation, coded each transcript independently. The overall inter rater agreement and kappa coefficients were, respectively 75% and 0.59 for the decision support skills and 76% and 0.68 for the communication skills categories. The frequency of DSAT skills coded: (a) were significantly correlated with three out of six patient and physician outcome measures (r>0.30, P<0.05); and (b) showed significant discrimination (P=0.05) or trends (P<0.15) in discrimination between the decision aid and pamphlet groups. The DSAT shows promise as a reliable and valid evaluation tool but requires further testing with larger samples. PMID- 12900094 TI - Informed choice: understanding knowledge in the context of screening uptake. AB - This study evaluates a scale measuring knowledge about a screening test and investigates the association between knowledge, uptake and attitudes towards screening. One thousand four hundred ninety-nine pregnant women completed the knowledge scale of the multidimensional measure of informed choice (MMIC). Three hundred forty-five of these women and 152 professionals providing antenatal care also rated the importance of the knowledge items. Item characteristic curves show that, with one exception, the knowledge items reflect a spread of difficulty and are able to discriminate between people. All items were seen as essential or helpful by both women and health professionals, with two items seen as particularly important and one as unimportant. There were some differences between health professionals, women with low risk results and women with high risk results. Knowledge was not associated with uptake, attitude, or the extent to which uptake was consistent with women's attitudes towards undergoing the test. PMID- 12900095 TI - Is anxiety a suitable measure of decision aid effectiveness: a systematic review? AB - Several trials have employed anxiety measures to assess decision aid effectiveness. This study employed a systematic review method to integrate their findings. The affective impact of decision aids and the appropriateness of anxiety as a measure of decision aid effectiveness are explored. From 11,361 citations generated from searching electronic databases and journals, 26 randomised controlled trials evaluated patient decision aids; 10 included anxiety measures (HADS; STAI). The data were too heterogeneous to integrate statistically. No studies showed an increase in anxiety from exposure to decision aids versus usual care. Some patterns emerged between level of anxiety and characteristics of the decision. As raised levels of anxiety are associated with both more effective decision strategies and stressful health interventions, anxiety is an inappropriate measure to employ when evaluating decision aids. Future research needs to investigate the relationship between affect, cognition and decision aids in order to facilitate effective patient decision making. PMID- 12900096 TI - Clinical decision-making at the end of life: the role of the patient's wish. AB - Information about medical decision-making in the last stage of life and the wishes of terminally ill patients is scarce. At two hospital departments, we observed 16 multidisciplinary meetings and recorded 110 discussions concerning 74 patients. Thirty-three of these discussions concerned either starting or forgoing potentially life-prolonging therapy or applying potentially life-shortening medical interventions. The wishes of the patients were considered in 15 of these discussions. In 6 of these 15 discussions, the patient's wish was unknown. In five discussions the decision was postponed to enable discussions with the patients. Potentially life-shortening decisions were never made if patients were known to disagree. No decisions about physician-assisted death were made. The study shows that medical end-of-life decisions are commonly shared between the involved caregivers and the patients. PMID- 12900097 TI - Integrated decision making: definitions for a new discipline. AB - The birth and early growth of a new discipline from the union of evidence-based medicine (EBM) and the theoretical foundations of human psychology and behaviour has led to a plethora of new terms to define and redefine a paradigm shift in clinical decision making. This paper provides a short glossary of (1) terms to describe the practitioner-patient relationship in decision making and (2) terms describing some of the tools for facilitating evidence-based decision making. In addition, it is proposed that a more acceptable name for this discipline may be 'integrated decision making' (IDM). A case is argued for the fact that this term better addresses many of the concerns raised by practitioners and consumers about barriers to the integration of evidence and patient preferences in clinical practice and the changing nature of the practitioner-patient relationship in healthcare today. PMID- 12900098 TI - Extending decision support: preparation and implementation. AB - The shift towards patient-centered care requires innovative, flexible decision making models. Current decision aids provide information and help patients clarify values. By focusing almost exclusively on these two components, the aids miss some necessary elements that can improve decision quality. We discuss two such elements in this paper. First is avoiding the tendency to move into providing information about options and outcomes before adequate preparation. The second is the tendency to neglect to provide adequate implementation support. PMID- 12900099 TI - Developing and using quantitative instruments for measuring doctor-patient communication about drugs. AB - Previous qualitative work has identified communication problems between doctors and patients in general practice consultations, particularly in relation to prescribing. This study aimed to develop quantitative measures to extend the research and provide instruments for both researchers and practitioners to use in monitoring communication and prescribing. Questionnaires were developed from existing instruments. When used with patients and doctors in a variety of general practices, the instruments appeared to be acceptable and had high response rates. While many consultations were satisfactory in terms of patients' expectations and their experiences with medicines, only 38% did not have any poor outcome. The results using quantitative instruments were similar to but less striking compared to our previous qualitative work. The research was developmental and findings suggest that unnecessary prescribing and problems in communication are more likely to lead to poor outcomes in terms of non-adherence and patients having barriers to using their medication. PMID- 12900100 TI - A qualitative study of physicians' perceptions of three decision aids. AB - The study objective was to investigate physicians' perceptions of three patient decision aids (DA). Semi-structured telephone interviews were conducted with 20 family physicians and 12 gynecologists about a DA for women considering long-term hormone replacement therapy; with 16 respirologists about a DA for the use of intubation and mechanical ventilation for patients with severe chronic obstructive pulmonary disease; and with 19 physicians (geriatricians, gastroenterologists, internists) about a DA for long-term placement of feeding tubes in the elderly. Participants were identified by a snowball sampling technique. The interviews were analyzed using standard qualitative methods. Most participants (81%) indicated some willingness to use the DAs. The characteristics of the DA viewed positively included it being: balanced, well organized, a useful tool, evidence-based, improves decision making process and multimedia. Some of the negative characteristics were stated as: too complex, the cost, the availability, only appropriate for certain groups of patients, and time consuming. The DAs were acceptable to most participants. Perceived positive and negative factors were similar for all DAs. Uptake of decision aids may be facilitated if physicians have an opportunity to examine and try them, and if they can have unfettered access to them for distribution purposes. PMID- 12900101 TI - Patient comprehension of information for shared treatment decision making: state of the art and future directions. AB - How does the way that information is presented potentially influence patients' consent for health treatments, in a shared decision making process? The goal of this paper is to present an overview of selected recent literature concerning patient health information presentation/use for treatment decision making. Recent work with patient populations has begun to extend early cognitive psychological work showing systematic biases in thinking. Key research findings are organized by type of format (probability, graphic, and qualitative/quantitative dimensions). The applied literature on this topic is amenable to only limited integration in regard to key findings, and relatively few novel approaches to improving information comprehension have been described in the health literature. Promising approaches being proposed, developed, and tested are described, such as enhanced-access computerized patient choice modules, "debiasing" techniques, and tailoring of information. Additional theoretical and practical issues are discussed, as well as selected policy implications of current knowledge. PMID- 12900102 TI - General practitioners' views on shared decision making: a qualitative analysis. AB - Despite rhetoric about increasing partnership in the consultation, shared decision making (SDM) is not necessarily happening in practice. As patient involvement in decision making cannot be achieved without general practitioners' (GPs) co-operation, attention to their concerns is essential. Previous research has used simulated patients to explore this area. Here, data from consultations and interviews with patients and GPs were used in focus group discussions enabling GPs to ground their views of the practical application of SDM in their own and other participants' real life practice. The following ideas were discussed; what constitutes SDM in practice, negotiation about treatment, perceptions of patients' viewpoints and perceived barriers to SDM. Agreement with SDM in theory may not translate into practice. The process outlined is being developed in order to establish whether the techniques could provide a supportive environment in which GPs can make changes to their practice to increase patient participation. PMID- 12900103 TI - Participation by clients and nurse midwives in family planning decision making in Indonesia. AB - In order to enhance understanding of the quality of decision making during family planning consultations in developing countries, provider competencies and client behaviors during 179 randomly selected consultations in Indonesia were assessed. Results show that family planning clients make a significant contribution to the quality of the decision-making process, most notably by identifying the problem requiring a decision, expressing their feelings about using a method, and asking questions. Client involvement may compensate for provider weaknesses, which tend to be in areas calling for interpersonal rather than technical skills. However, the programmatic ideal of informed choice has not yet been realized. Supervisors, trainers, communicators, and program managers can improve the quality of decision making by: creating opportunities for client involvement during consultations, strengthening providers' ability to fully inform clients about their options, and making providers aware of the opportunities for decision making in consultations with continuing clients. PMID- 12900104 TI - Incorporating patients' views and experiences of life with IBS in the development of an evidence based self-help guidebook. AB - To become active partners in chronic illness management, patients need relevant information which clearly explains treatment choices. This paper describes a method of producing information to feed into production of a guidebook for people with irritable bowel syndrome (IBS). Users were involved throughout the development process. Five focus groups were held involving 23 people with IBS out of 147 who responded to a newspaper article asking for help with a study of self care in IBS. Qualitative analysis of transcripts from these meetings formed the basis of guidebook development. Patients described their experiences, coping strategies, experiences of the healthcare system, treatments and social consequences of the condition. Medical literature searches were made to find answers to identified areas of information need. The book includes direct quotes from patients describing their own experiences. Guided and practical ways of support are required for people with IBS who want to self-manage their condition. PMID- 12900105 TI - The development of COMRADE--a patient-based outcome measure to evaluate the effectiveness of risk communication and treatment decision making in consultations. AB - Some instruments have been developed to evaluate 'decision effectiveness' such as the 'satisfaction with decision (SWD)' and 'decision conflict (DCS)' scales and are validated for the US context. Patients identify further outcome domains that are not fully covered in these scales. We developed a patient-based outcome measure to evaluate risk communication and decision making effectiveness, addressing these further domains and validated for use in the UK. Formulation of items was based on literature review and key informant interviews. The instrument was piloted and developed (including psychometric assessment) in the context of a trial of decision making and risk communication interventions in general practice. Patients were recruited to the trial with known atrial fibrillation, prostatism, menorrhagia or menopausal symptoms. High response rates (96% in first phase) indicated acceptability to patients. A 20 item patient-based outcome measure, with two sub-scales for 'risk communication' and 'confidence in decision', was produced. It allows for paternalistic, shared or informed choice decision making models. Usage in further studies will facilitate systematic reviews. Consequently, lessons for wider application of these interventions in practice, and policy implications regarding greater patient involvement should then be clearer. PMID- 12900106 TI - Understanding why decision aids work: linking process with outcome. AB - Decision aids help patients make treatment choices. There is little empirical evidence to explain how they work. The results from this randomised controlled trial comparing routine with decision-aided consultations in the prenatal diagnosis for Down's syndrome context are used to describe the strategies employed during decision making, to assess the impact of a decision aid on decision processes, and to investigate decision process and outcome associations. Data were elicited from two content analyses of consultation transcripts and questionnaires assessing knowledge, anxiety, decisional conflict, reasons, and information usefulness. 68/106 women completed measures at consultation and follow-up. Decision-aided women employed more cognitive and emotional strategies during decision making. More negative evaluations during decision making were associated with better outcomes. Decision-aided consultations facilitated the employment of strategies associated with more effective choices. These consultations take longer and elicit greater expressions of negative affect, so may be less rewarding encounters for health professionals. PMID- 12900108 TI - Prognostic factors and treatment of penetrating gunshot wounds to the head. AB - BACKGROUND: In an attempt to evaluate the predictive factors of morbidity and mortality in patients who suffered from civilian gunshot wounds to the head, we reviewed a series of 319 patients admitted to the Hospital Santa Marcelina, Sao Paulo, Brazil, between 1994 and 2000. METHODS: Clinical and cranial computed tomography (CT) results are described. The initial Glasgow Coma Scale (GCS), the presence of an unilateral dilated pupil or medium fixed pupils, and five different findings in the CT scan were considered as variables. The Qui-Square Fisher test was utilized to verify the correlation between the presence of the variables and the occurrence of an increased mortality rate and of an unfavorable outcome (Glasgow Outcome Scale = 2 and 3). RESULTS: In 265 cases the missile penetrated the dura (83%). In our study there was a significant correlation between the low GCS scores on admission and a higher mortality (p < 0.001). This kind of correlation was also noted with patients admitted with unilateral dilated pupil and medium fixed pupil. There were 187 patients (70.5%) evaluated by CT scan. There was a significant correlation between the presence of transventricular or bihemispheric central type trajectory and high mortality. The patients admitted with unilobar supratentorial wounds resulted in better outcome when compared to those with bilobar or multilobar wounds (p < 0.001). A group of 156 patients was submitted to an aggressive surgical protocol. The best results were seen in patients admitted with initial GCS score higher than 8. CONCLUSIONS: We conclude that low GCS scores at admission, unilateral dilated pupil or medium fixed pupil, transventricular or bihemispheric central type trajectory, and bilobar or multilobar wounds noted through CT scan are predictive factors of high morbidity and mortality in patients with gunshot wounds to the head, in our clinical experience. We also conclude that surgical treatment is not recommended for patients with penetrating wounds and GCS score of 3 to 5 in the absence of hematoma causing a mass effect. PMID- 12900112 TI - The treatment of wartime brain wounds: traditional versus minimal debridement. AB - BACKGROUND: Since World War II, surgeons in Western armies dealing with brain wounds have generally advocated thorough missile track debridement, and many have urged meticulous dural closure to prevent cerebral spinal fluid (CSF) leakage and subsequent infection. For the last decade some reports have appeared wherein wartime brain wounds have been treated by minimal brain debridement with little or no attention to tight dural closure. This report compares and contrasts postdebridement complications reported with each method. METHODS: I reviewed the pertinent neurosurgical papers from Vietnam, the Israeli war in Lebanon, and the Iran-Iraq war and compared the results of traditional debridement and water-tight dural closure to those following minimal debridement without close attention to dural closure. RESULTS: Minimal brain debridements without tight dural closure were up to 20 to 30 times more likely to require additional debridement, had a 10 15 times higher incidence of life-threatening CSF leakage, a fivefold increase in postdebridement meningitis, and a 2.5 times greater risk of fatal meningitis. CONCLUSIONS: Compared to thorough brain debridement and watertight dural closure, minimal debridements and nonwatertight dural closure give inferior results. PMID- 12900110 TI - Minimal debridement or simple wound closure as the only surgical treatment in war victims with low-velocity penetrating head injuries. Indications and management protocol based upon more than 8 years follow-up of 99 cases from Iran-Iraq conflict. AB - BACKGROUND: During military conflicts, surviving victims traditionally underwent large enough craniectomy or craniotomy to achieve extensive debridement of the in driven bone, shell fragments, and debris. Meanwhile, the completely as well as the partially devitalized adjacent brain tissue would also be debrided. This might have led to additional neurologic deficit or lesser degree of recovery of functions. METHOD: We studied the efficacy and safety of much more limited or even absence of surgical intervention in a selected group of wartime head-wounded patients. Among 1,150 war victims with penetrating head wounds referred to two major centers affiliated with Tehran University of Medical Sciences during the 8 year period spanning the Iran-Iraq conflict, there were 191 head-wounded patients who underwent either no surgical intervention or a very limited debridement. We have attempted to follow up as closely as possible for more than 8 years. Ninety nine of them fulfilled the criteria to be included in this series. Close follow up for development of cerebral infections, functional recovery, or development of new neurologic deficits and epilepsy were performed. At least 3 contrast-enhanced CT scans were taken for each patient. All patients were victims of the battle field areas with low- to moderate-velocity missile or shell fragment injury. The Glasgow Coma Scale (GCS) scores of the patients were between 8-14 when visited by the first attending neurosurgery staff. The factors considered to be effective in the outcome analysis were the entry point, the number of the in-driven bone fragments, the retained shell fragments, the brain compartments affected, and the paranasal sinus or skull base involvement with or without cerebrospinal fluid (CSF) fistula. RESULTS: There were 13 unilobar, 44 bilobar, 4 trilobar, 38 transventricular, and 3 skull base lesions. The number of retained metal fragments varied between 1 to 6. The number of retained bone fragments varied between 1 to 5 in 73 victims and more than five chips in other cases. The outcome was good (back to work) in 90 patients and poor (severely disabled) in 4. Five patients died because of severity of brain damage and meningitis. The effect of the different enumerated variables upon the outcome was measured using chi(2) and Fisher exact test, which was nonsignificant for all of the variables except for the orbitofacial entry point of injury (p = 0.00006). CONCLUSION: This study seems to indicate that not only is reoperation for retained bone or shell fragments unnecessary, but surgeons having modern neuroradiological instruments available to detect the proper explanation for changes in GCS of the patients may decide not to proceed with any surgical intervention in a good number of patients. As a result of the "no surgical treatment protocol," one can preserve cerebral tissue without exposing the patient to undue risk of seizures, infection, or death by leaving behind some or all the in-driven bone and shell fragments. PMID- 12900113 TI - Penetrating head wound: a remarkable case. AB - BACKGROUND: The modern management of penetrating head injuries is generally considered a neurosurgical specialty that relies heavily on the use of sequential imaging techniques, an ever-changing armamentarium of sophisticated antibiotics and specific neurosurgical operative skills. Unfortunately these optimal therapeutic components are frequently not available to patients injured in underdeveloped countries. METHODS: An unusual case of a young patient suffering a penetrating brain wound and undergoing delayed treatment in an African mission hospital is reviewed. RESULTS: A functional but neurologically impaired outcome resulted from limited surgical debridement and short-term broad-spectrum antibiotic administration. CONCLUSIONS: The multiple exigencies of surgical practice in an underdeveloped African nation do not preclude successful management of penetrating head trauma. PMID- 12900115 TI - Risk factors for outcome and complications of dorsal foraminotomy in cervical disc herniation. AB - BACKGROUND: Dorsal foraminotomy is a standard operative procedure for lateral cervical disc herniation. Factors associated with surgical complications and clinical outcome in dorsal foraminotomy of cervical disc herniation were evaluated in a retrospective cohort study. METHOD: Thirty-nine patients were operated upon for unilateral, monosegmental, mediolateral cervical disc herniation (+/- associated spondylosis) from 1997 to 1999. Preoperative radiologic imaging and surgical reports were analyzed. Motor disfunction, neck irritation, and radicular pain were evaluated. Outcome was ranked according to modified Odom's criteria at 6 weeks and 1 year postoperatively. RESULTS: Six weeks after injury 7 of 39 patients (18%) showed neck irritation. No new neurologic deficit was seen. All patients with preoperative paresis improved; two had early relapses of a medial soft disc prolapse (2/39). Residual radicular pain was seen in 3 of 39 patients (8%) within 30 days postoperatively, necessitating surgical revision. Factors of surgical failure were associated spondylosis (2/3) and residual mediolateral disc protrusion (1/3). In one patient with associated spondylosis, local pain due to a symptomatic fracture of the lateral process of D1 resolved after revision. Duration of preoperative radicular pain was identified as a risk factor for unfavorable outcome. CONCLUSION: In lateral cervical disc herniation, associated spondylosis or medial disc protrusion poses a significant risk of surgical failure and complications of dorsal foraminotomy. Reducing the radicular failure rate by enlarging the bony decompression may lead to local failure. In well-selected patients with a lateral cervical free disc fragment, dorsolateral foraminotomy is successful and safe. PMID- 12900118 TI - Square cervical laminoplasty incorporating spinous process: surgical technique. AB - BACKGROUND: We have used a laminoplasty technique in multilevel cervical canal stenosis that incorporates spinous process in the neural arch and does not require free graft or foreign material. METHODS: In this technique, laminae and spinous processes were used to enlarge stenotic cervical spinal canal. Three patients (mean age 34.6 years) formed the study group with a mean follow-up period of 24.6 months. Postoperative computed tomography (CT) and lateral radiographs were used to assess results in terms of bony union, canal diameter, and alignment of cervical spine. RESULTS: All patients noted some improvement in both sensory and motor functions. The average increase in sagittal diameter of cervical spinal canal was 4.2 mm, and decrease in range of motion (ROM) was 13.2 degrees. Bony fusions at the gutters were seen after 6 months. CONCLUSION: This new technique of cervical laminoplasty is safe, effective, relatively easy, and avoids complications related to free grafts or metallic or nonmetallic foreign materials used in other techniques PMID- 12900120 TI - Multiple aneurysms. PMID- 12900121 TI - A case of giant expanding cephalhematoma: does the administration of blood coagulation factor XIII reverse symptoms? AB - BACKGROUND: The mechanism of continuous massive hemorrhage into the subperiosteal space in children without coagulation defects is unknown. We report a case of giant expanding cephalhematoma reversed by the administration of blood coagulation factor XIII concentrate. METHOD: The patient was an 8-year-old boy with a history of minor head trauma who developed a giant expanding cephalhematoma with intraorbital extension. The laboratory data showed severe anemia, but a routine blood coagulation test showed no abnormalities except for a low factor XIII level. RESULT: The administration of factor XIII concentrate completely reversed the symptoms in 2 weeks. CONCLUSION: We speculate that one of the possible mechanisms of cephalhematoma expansion without blood coagulation defects might be acquired factor XIII deficiency from severe hemorrhage in a hematoma. PMID- 12900124 TI - Clinical outcomes in patients with Chiari I malformation: a review of 27 cases. AB - BACKGROUND: Chiari I malformation is a congenital maldevelopment defined as downward herniation of the cerebellar tonsils through the foramen magnum. It has been treated using a variety of surgical procedures. Surgical outcomes have been reported with varying results throughout the literature. METHODS: We retrospectively reviewed 27 patients with Chiari I malformation operated at the University Hospital during a 9-year period from 1988 to 1997. We assessed preoperative and postoperative signs and symptoms in all the patients including headache, neck pain, nystagmus, vertigo, weakness, spasticity, atrophy, numbness, pain and temperature dissociation, diplopia, dysphagia, and sphincter dysfunction. Each patient was analyzed to determine if there was an arrest in the progression of the disease after surgical intervention. RESULTS: Syringomyelia was present in 59% of the patients. Only 1 patient who presented with neck pain improved. One patient reported new onset headache, and one patient described his headache resolved. Vertigo resolved in three patients; two patients stated mild improvement, and one patient reported worsening. Nystagmus improved or resolved in six patients. Weakness improved in only two patients who did not have syringomyelia. Dysphagia improved in two patients, and in the others it remained unchanged. Diplopia, spasticity, atrophy, and numbness remained unchanged. CONCLUSIONS: Our study provides evidence that the main benefit of the surgical management in patients with Chiari I malformation with or without syringomyelia is to arrest the progression of the disease. PMID- 12900126 TI - Percutaneous trigeminal ganglion balloon compression for treatment of trigeminal neuralgia, part II: results related to compression duration. AB - OBJECTIVE: The purpose of this study is to find out how different compression time affects the results of percutaneous trigeminal ganglion compression for treatment of trigeminal neuralgia. METHODS: This study includes 80 patients with intractable third-branch trigeminal (V3) neuralgia who had received percutanious trigeminal ganglion balloon compression. All the patients received some treatment protocol except for the duration of compression. Group 1 patients received 60 second and group 2 patients received 180-second compression. A computerized pressure recording system was used for pressure monitoring and analysis. RESULTS: Both groups had 100% immediate pain relief and all patients experienced mastication weakness immediately after the procedure. The facial numbness was severe in group 2 in the first trigeminal (V1) distribution (p<0.05) but not in the second or third trigeminal (V2,3) distribution (p>0.05) in the immediate period after the operation. The recurrence rate in the first year follow-up was higher in group 1 (5%) than in group 2 (2.5%) but did not reach the statistical difference (p>0.05). At 1 year follow-up after the procedure, group 1 and lower incidence of facial numbness over all trigeminal distribution (V1,2,3) than group 2 (p<0.05) mastication weakness all recovered during the first-year follow up. CONCLUSIONS: With accurate monitoring of the balloon pressure during the percutaneous trigeminal ganglion compression, it was found that the shorter duration of compression had less side effect. At one year follow-up, the incidence of recurrrence rate was slightly higher in the patients who received 60 second compressions than in those who received 180-second compressions, but there was no significant statistical difference. Whether patients with first or second branch of trigeminal neuralgia require longer compression duration needs further study. PMID- 12900129 TI - Frontal sinus repair with free autologous bone grafts and fibrin glue. AB - BACKGROUND: The authors describe a technique for repairing the frontal sinus with autologous bone grafts removed during craniotomy and fibrin glue. METHODS: This technique was used in 12 patients who underwent craniotomy for aneurysms (n = 9), brain tumors (n = 2), and acute epidural hematoma (n = 1). RESULTS: The repair was successful in all cases. There were no instances of postoperative infection or leakage of cerebrospinal fluid. No complications from the repair have occurred over a mean follow-up of 51 months. CONCLUSION: This technique is simpler than others used to seal a frontal sinus. PMID- 12900131 TI - Brain and systemic temperature in patients with severe subarachnoid hemorrhage. AB - BACKGROUND: The significance of brain temperature (BT) in patients with severe brain damage remains unclear. This study investigated the relationships between BT, systemic temperature (ST), and clinical outcome in patients with severe subarachnoid hemorrhage. METHODS: Thirty-one comatose patients with severe subarachnoid hemorrhage underwent ventricular drainage immediately after admission. The ventricular catheter also allowed monitoring of BT. ST was continuously measured using a bladder catheter with thermistor probe. RESULTS: BT at the start of the monitoring was lower than ST in four patients, and all died of brain swelling. BT was higher than ST at first but later fell below ST ("temperature reversal") in 12 patients, who all died of acute brain swelling. BT was higher than ST throughout the monitoring in 15 patients. Five of these patients died of causes other than brain swelling such as rerupture of the cerebral aneurysm, multiple organ failure, or respiratory failure. The other 10 patients survived with various degrees of disability. CONCLUSIONS: Observation of BT and ST can predict the outcome of severe subarachnoid hemorrhage. PMID- 12900133 TI - Safety of magnesium-lidocaine combination for severe head injury: the Turin lidomag pilot study. AB - BACKGROUND: Neuroprotection in the setting of severe head injury (SHI) remains an unsettled problem. We tested a combination of high-dose magnesium and low-dose lidocaine, infused over 3 days, in a pilot study to assess safety. This combination appears indicated to protect both gray and white matter from secondary injury following SHI. METHODS: We studied 32 consecutive patients admitted to the emergency department of our hospital, a large tertiary referral center. RESULTS: No toxicity was observed. Mortality was lower than published statistics. CONCLUSIONS: These results open the stage to a controlled randomized study. PMID- 12900135 TI - En Bloc removal of soft tumors within the brain parenchyma using spoon retractors: traction-dissection method. Technical note. AB - BACKGROUND: En bloc removal of soft tumors within the brain parenchyma has rarely been performed. We describe a safe technique for en bloc removal of the tumors using a spoon retractor, which enables retraction while holding the soft mass upward. METHODS: Dissection of a tumor mass is performed under traction of the surrounding brain tissue by retracting the mass using a spoon retractor. A dissection plane is first established in a relatively safe area, not opposite the critical area, after a corticotomy. The dissection plane is then extended toward the critical side. After circumferential dissection of the tumor mass, the dissection is continued spirally into deeper regions by retracting and holding the mass upward using a spoon retractor. In this way, the tumor is removed en bloc. CONCLUSIONS: The traction-dissection method using spoon retractors is useful in performing en bloc removal of soft tumors within the brain parenchyma less invasively and provides an appropriate operating field even at depth, reducing intraoperative bleeding, and in vascular rich tumors, possibly preventing tumor seeding in cases of malignant tumors. PMID- 12900136 TI - The mystery of gold foil. PMID- 12900137 TI - Whatever happened to dreaming? PMID- 12900138 TI - Primary empty sella syndrome. PMID- 12900139 TI - Modeling transmission of directly transmitted infectious diseases using colored stochastic Petri nets. AB - In order to improve our understanding of directly transmitted pathogens within host populations, epidemic models should take into account individual heterogeneities as well as stochastic fluctuations in individual parameters. The associated cost results in an increasing level of complexity of the mathematical models which generally lack consistent formalisms. In this paper, we demonstrate that complex epidemic models could be expressed as colored stochastic Petri nets (CSPN). CSPN is a mathematical tool developed in computer science. The concept is based on the Markov Chain theory and on a standard well codified graphical formalism. This approach presents an alternative to other computer simulation methods since it offers both a theoretical formalism and a graphical representation that facilitate the implementation, the understanding and thus the replication or modification of the model. We explain how common concepts of epidemic models--such as the incidence function--can be easily translated into an individual based point of view in the CSPN formalism. We then illustrate this approach by using the well documented susceptible-infected model with recruitment and death. PMID- 12900140 TI - Global dynamics of an SEIR epidemic model with saturating contact rate. AB - Heesterbeek and Metz [J. Math. Biol. 31 (1993) 529] derived an expression for the saturating contact rate of individual contacts in an epidemiological model. In this paper, the SEIR model with this saturating contact rate is studied. The basic reproduction number R0 is proved to be a sharp threshold which completely determines the global dynamics and the outcome of the disease. If R0 < or =1, the disease-free equilibrium is globally stable and the disease always dies out. If R0 > 1, there exists a unique endemic equilibrium which is globally stable and the disease persists at an endemic equilibrium state if it initially exists. The contribution of the saturating contact rate to the basic reproduction number and the level of the endemic equilibrium is also analyzed. PMID- 12900141 TI - Analyzing bioterror response logistics: the case of smallpox. AB - To evaluate existing and alternative proposals for emergency response to a deliberate smallpox attack, we embed the key operational features of such interventions into a smallpox disease transmission model. We use probabilistic reasoning within an otherwise deterministic epidemic framework to model the 'race to trace', i.e., attempting to trace (via the infector) and vaccinate an infected person while (s)he is still vaccine-sensitive. Our model explicitly incorporates a tracing/vaccination queue, and hence can be used as a capacity planning tool. An approximate analysis of this large (16 ODE) system yields closed-form estimates for the total number of deaths and the maximum queue length. The former estimate delineates the efficacy (i.e., accuracy) and efficiency (i.e., speed) of contact tracing, while the latter estimate reveals how congestion makes the race to trace more difficult to win, thereby causing more deaths. A probabilistic analysis is also used to find an approximate closed-form expression for the total number of deaths under mass vaccination, in terms of both the basic reproductive ratio and the vaccination capacity. We also derive approximate thresholds for initially controlling the epidemic for more general interventions that include imperfect vaccination and quarantine. PMID- 12900142 TI - The effect of heterogeneity on optimal regimens in cancer chemotherapy. AB - Optimal drug regimens for cancer chemotherapy are determined when knowledge is only available on the behaviour of the tumour and the drugs used, over a population of patients. The case of two drugs is investigated where they are equivalent on average. Our calculations indicate that the optimal regimen has both drugs given initially but then sequences the two drugs. Our calculations also indicate that as tumour heterogeneity increases, the benefit to be gained from the optimal regimen can decrease in comparison to reasonable regimens. This has the effect of complicating the calculation of optimal regimens in a clinical setting, and may explain why results in experimental oncology fail to carry over to clinical oncology. PMID- 12900143 TI - On vaccine efficacy and reproduction numbers. AB - We consider the impact of a vaccination programme on the transmission potential of the infection in large populations. We define a measure of vaccine efficacy against transmission which combines the possibly random effect of the vaccine on individual susceptibility and infectiousness. This definition extends some previous work in this area to arbitrarily heterogeneous populations with one level of mixing, but leads us to question the usefulness of the concept of vaccine efficacy against infectiousness. We derive relationships between vaccine efficacy against transmission, vaccine coverage and reproduction numbers, which generalize existing results. In particular we show that the projected reproduction number RV does not depend on the details of the vaccine model, only on its overall effect on transmission. Explicit expressions for RV and the basic reproduction number R0 are obtained in a variety of settings. We define a measure of projected effectiveness of a vaccination programme PE=1-(RV/R0) and investigate its relationship with efficacy against transmission and vaccine coverage. We also study the effective reproduction number Re(t) at time t. Monitoring Re(t) over time is an important aspect of programme surveillance. Programme effectiveness PE is less sensitive than RV or the critical vaccination threshold to model assumptions. On the other hand Re(t) depends on the details of the vaccine model. PMID- 12900144 TI - Global context, generic tools. PMID- 12900145 TI - Science, politics and animal health policy: epidemiology in action. AB - Public policy decisions underlie society's response to current animal health issues ranging from emerging diseases and public health threats to food safety concerns and sustainable animal agriculture strategies. Despite strong calls for "science-based" decisions, animal health policy most commonly emerges at the interface of science and politics. Too often scientists' disdain for politics limits their involvement in formulating policy. By contrast, epidemiologists are ideally qualified to bring scientific skills to complex policy issues through analytical, macro-epidemiological approaches that consider the economic, legal, and cultural context of policy issues as well as the biological and medical aspects. Risk analysis provides a systematic approach to evaluating animal health issues and comparing policy options. Capturing these opportunities for applied epidemiology requires an understanding of the policy-making process as well as the basic principles of epidemiology. Furthermore, epidemiology training programs must incorporate communications skill building and experiential learning opportunities in a team environment. PMID- 12900146 TI - Associations between dogs that were serologically positive for Rickettsia conorii relative to the residences of two human cases of Mediterranean spotted fever in Piemonte (Italy). AB - A geographic information system and K-function analysis were used to evaluate the spatial association of canine serological results for Rickettsia conorii, the causative agent of Mediterranean spotted fever (MSF), and clinical cases of MSF in humans in Piemonte, northwest Italy. The residences of dog owners were clustered in two rural villages in the province of Cuneo, where two human cases of MSF occurred in 1997 and 1998. Eighteen out of 116 dogs examined were positive by indirect immunofluorescent assay (IFA+, titre > or =1:160) for MSF. K functions were compared for IFA+ dogs and for all dogs sampled. Monte Carlo and bootstrap simulations demonstrated that clustering of IFA+ dogs was significantly greater than clustering of all dogs, at distances of less than 0.6 km from human cases of MSF. Logistic regression analysis indicated that the risk of being IFA+ was highest for dogs residing within the first quartile of distance (0.7 km) from human cases of the disease, and for dogs that were not confined. However, year and season of blood collection were not associated with IFA status. It was concluded that a relatively high dog population density along with a rural or semi-rural environment favours the occurrence of emergent foci of MSF in the province of Cuneo. PMID- 12900147 TI - Risk analysis of the spread of classical swine fever virus through "neighbourhood infections" for different regions in Belgium. AB - Risk factors associated with the occurrence of "neighbourhood infections" [Epidemiology of classical swine fever. In: Truszczynski, M. (Ed.), Proceedings of the Workshop on Diagnostic Procedures and Measures to Control Classical Swine Fever in Domestic Pigs and the European Wild Boar. Pulaway, Poland, pp. 119-130] during classical swine fever (CSF) outbreaks were examined based on information collected during a CSF-epidemic, which occurred in the East Flanders Province of Belgium in 1994. The only risk factor that was associated with the occurrence of "neighbourhood infections" was a kernel estimation of the intensity of neighbouring herds (P=0.055) [Interactive spatial data analysis. Pearson Education Limited, Harlow, Essex], i.e. the higher the kernel estimation, the higher the risk for the occurrence of neighbourhood infections. In a second part of the study, the likelihood for the occurrence of neighbourhood infections within an area with a 1 km radius was predicted for every Belgian pig herd, assuming that the herd was infected with CSF-virus. For the prediction of these likelihoods, the model resulting from the risk assessment was used. Finally, the predicted likelihoods were transformed into a raster map after applying a smoothing technique. As a result, different areas in Belgium of higher or lower risk for CSF-virus spread through "neighbourhood infections" could be identified on the map. The areas in Belgium where CSF-outbreaks including "neighbourhood infections" occurred in the past decades were all predicted by the model to be of high risk. PMID- 12900148 TI - A review of porcine reproductive and respiratory syndrome virus in Dutch breeding herds: population dynamics and clinical relevance. AB - Understanding the spread of porcine reproductive and respiratory syndrome virus (PRRSV) in pig populations is essential to the development of effective PRRS prevention and control strategies. Moreover, knowledge of the field dynamics of PRRSV in pigs will provide insights into the clinical relevance of PRRS, and will enable the targeting of interventions. This review of PRRSV includes discussion on the occurrence of outbreaks, the persistence of infection and the fade-out of infection in Dutch breeding herds. The dynamic character of PRRSV infections in endemically infected herds and the relevance of the disease under Dutch field conditions are also highlighted. Furthermore, several strategies aimed at controlling the spread of PRRSV are discussed. PMID- 12900149 TI - Design and analysis of an Actinobacillus pleuropneumoniae transmission experiment. AB - This paper describes a methodology to quantify the transmission of Actinobacillus (A.) pleuropneumoniae from subclinically infected carrier pigs to susceptible contact pigs, and to test the effect of possible interventions on the transmission. The methodology includes the design of a transmission experiment, and a method with which A. pleuropneumoniae transmission can be quantified and with which the effect of an intervention on the transmission can be tested. The experimental design consists of two parts. First, subclinically infected carrier pigs are created by contact exposure of specific-pathogen-free pigs to endobronchially inoculated pigs. Second, transmission is observed from the group of carrier pigs to a second group of susceptible contact pigs after replacing the inoculated pigs by new contact pigs. The presented analytical method is a generalised linear model (GLM) with which the effect of an intervention on the susceptibility and infectivity can be tested separately, if the transmission is observed in heterogeneous populations. The concept of the experimental transmission model is illustrated by describing an A. pleuropneumoniae transmission experiment in which the effect of vaccination on the susceptibility is quantified. Although it could not be demonstrated that vaccination has an effect on the susceptibility of pigs, it was demonstrated that nasal excretion of A. pleuropneumoniae is related to the infectivity of pigs. PMID- 12900150 TI - Development of the Dutch Johne's disease control program supported by a simulation model. AB - The development of a simulation model, "JohneSSim", was part of a research program aimed at designing a national Johne's disease control program for The Netherlands. Initially, the focus was mainly directed towards different compulsory "test-and-cull" strategies. However, the results from the JohneSSim model showed that eradication of Johne's disease based on such strategies would not be possible within 20 years and that it was also economically unattractive. However, improved calf management seemed to be more effective in reducing the prevalence within the same time period. Simulation of a strategy using an "ideal test" (80% sensitivity in all infected animals) showed a considerably faster decrease in prevalence. However, this strategy proved to be economically unattractive because of the high culling rate of (young) test-positive animals. The simulation model was also adapted to study beef cow herds. However, the results indicated that none of the strategies were able to reduce the mean true prevalence to almost zero for such herds. Only strategies based on "separation of calves and adult animals" proved to significantly reduce the prevalence but such a strategy is unpractical and uneconomic for Dutch beef cow herds. Due to this finding and the relative low number of Dutch beef cow farms, first priority has been given to the development of a Johne's disease control program for dairy farms. Based on the results of the "JohneSSim" model, the new national voluntary Johne's disease control program for dairy, Paratuberculosis Program Netherlands (PPN), started in September 2000. The PPN is based on a stepwise improvement of calf hygiene, with little dependency on "test and culling". The model results indicated that, if dairy farmers consistently carried out the necessary management adaptations, PPN considerably decreased prevalence and that it was economically more attractive than any previous plans. PMID- 12900151 TI - Controlling Echinococcus multilocularis-ecological implications of field trials. AB - Two field trials to reduce the prevalence of Echinococcus multilocularis in foxes have been conducted in recent years. Although both trials reduced prevalence considerably, they failed to eradicate the parasite in the study region. Following the control trial in northern Germany, prevalence recovered unexpectedly and rapidly, reaching pre-control levels five quarters (15 months) after the end of control. To understand the internal dynamics of the parasite host system's reaction to control, we developed a spatially explicit simulation model, Echi. The simulation model incorporates the information available concerning fox tapeworm population dynamics. Using epidemiological parameters to adjust pre-control prevalence, the model predicts the temporal evolution of the prevalence of E. multilocularis in controlled foxes without departing from the range of uncertainty of the field data. However, the model does not predict the rapid pre-control recovery observed in the field trial. The deviation of the model's prediction from field data indicates the involvement of processes not yet taken into account. We modified the model step by step to mimic processes with the potential to cause the rapid post-control recovery of the prevalence of E. multilocularis in foxes. Neither the longevity of tapeworm eggs nor the migratory behaviour of foxes showed any influence on the post-control reaction of the parasite-host system. However, landscape structures leading to a heterogeneous distribution of infected foxes have the potential to alter the system's reaction to control. If infected foxes are concentrated in multiple clusters in the landscape, the model prediction tallied with the range of uncertainty of the field data. Such spatial distribution of infected foxes may be caused by differential abiotic conditions influencing the survival of tapeworm eggs. The model was found to comply best with field data if the foxes acquire partial immunity by being exposed to the fox tapeworm. Both hypotheses explaining the rapid post-control recovery of the prevalence of E. multilocularis observed in the fox population were supported by field data. Both hypotheses have far reaching consequences for future control trials. The spatial aggregation of infected foxes would enable control efforts to be concentrated on these highly infected areas. However, the acquisition of immunity acts as a buffer to control, necessitating intensified control measures. PMID- 12900152 TI - A case control study of factors and infections associated with clinically apparent respiratory disease in UK Thoroughbred racehorses. AB - A matched case control study was used to determine infections and other factors associated with clinically apparent respiratory disease in young racehorses in training in the UK. A total of 170 cases, defined as horses with sudden onset coughing, nasal discharge or pyrexia, were identified and matched to 632 non affected controls by trainer and time period. Factors examined included age, sex, time since entry into the training yard, time since last race and different infections including tracheal and nasopharyngeal (NP) bacteria and viruses. Multivariable conditional logistic regression (CLR) modelling was used to evaluate the risk of being a case for variables after adjustment for other factors. Three analyses were conducted using clinical cases as outcomes, which were compared with: (i) controls without evidence of subclinical inflammatory airway disease (IAD) (ii) controls with evidence of subclinical IAD and (iii) all controls irrespective of IAD status. A fourth analysis was conducted comparing the two groups of controls, i.e. those with and without IAD. Younger horses and those that had entered training more recently were at increased risk of suffering episodes of clinically apparent respiratory disease. Among the infections, increasing numbers of Pasteurella/Actinobacillus spp. in tracheal washes were associated with increasing risk of clinical disease. Tracheal infection with Streptococcus zooepidemicus was associated with both clinical respiratory disease and subclinical IAD when compared with controls with no evidence of IAD. This explained the lack of association between clinical cases and S. zooepidemicus when all controls were used. Tracheal isolation of Mycoplasma felis was also associated with clinical disease after controlling for other factors. An inverse association was identified between risk of clinically apparent disease and isolation from tracheal washes of the transient, non-pathogenic bacteria Staphylococcus and Acinetobacter spp. There was no significant association identified between clinical disease and infection with equine herpesviruses-1 and -4, rhinoviruses-1 and -2 or adenovirus. Equine influenza was significantly associated with clinical respiratory disease but it was a very rare infection in this well-vaccinated population, only occurring in three cases. PMID- 12900153 TI - Open status of pig-breeding farms is associated with slightly higher seroprevalence of F18+ Escherichia coli in northern Belgium. AB - F18+ enterotoxigenic and verotoxigenic Escherichia coli (E. coli) are widespread and induce post-weaning diarrhoea and oedema disease, respectively. No commercial vaccine exists. Therefore, the F18 herd seroprevalence of 178 randomly selected pig-breeding farms in northern Belgium was evaluated by determining F18-specific IgG antibody titres in pooled sera of sows per breeding farm. This survey reported the first data concerning F18 herd seroprevalence and showed that F18+ E. coli seroprevalence was common in northern Belgium. Significantly more open pig-breeding farms were F18 seropositive (96.4%) compared to closed (88.8%). Furthermore, no clear association was determined between the F18 herd seroprevalence and the number of sows per breeding farm (rsp = 0.19), or with the number of fattening pigs per breeding farm (rsp = 0.17). PMID- 12900154 TI - Perceived constraints to private veterinary practice in India. AB - We used an exploratory study in two districts of West Bengal State, India to find out the constraints of private veterinary practice. Privatization of veterinary services is being seen as an alternative to government-sponsored veterinary services to livestock. In December 2000 to February 2001, 50 private veterinary practitioners were interviewed from the districts. Their opinion was that weak regulations about competition from illegal practitioners and the use of veterinary drugs, competition by animal-health technicians (who are likely to charge less than veterinarians), absence of efficient diagnostic-laboratory support, and lack of commercial livestock farming were the main constraints faced. PMID- 12900155 TI - Single versus double testing of meat-juice samples for Salmonella antibodies, in the Danish pig-herd surveillance programme. AB - In Denmark, a national serological surveillance-and-control programme for Salmonella in pigs has been in operation since 1995. The programme is based on the Danish mix-ELISA and uses double testing (two ELISA-wells used per sample) of meat-juice samples taken in relation to slaughter. All herds are classified monthly into one of the three levels; the classification is based on the percentage of positive serological results in the previous 3 months. In connection with evaluation of the programme in 2001, we investigated whether single testing (testing in one well only) could be expected to be sufficiently precise compared to double testing. Data from the year 2000 were used, and mathematical modelling. Single testing was simulated by randomised selection of one of the two results in the double testing. A slight increase in the prevalence of Salmonella-positive samples (1.02-1.09 times more through the four quarters of the year 2000) was found in the simulated single testing, as compared to the double testing. Around 0.5% of the herds would be allocated to another herd level in single testing-almost equal numbers one level up and one level down. No herd being seronegative in double testing would be allocated to levels 2 or 3 (herds with >40 or >70%, respectively, serological reactors) in single testing. The prevalence of "false-positive" diagnoses (positive in single testing and negative in double testing) and inversely defined "false-negative" diagnoses varied from 4.2 to 8.7% and from 3.2 to 4.5%, respectively, through the four quarters of the year 2000. The probability of allocating a herd to a wrong level due to sampling error was on the average 6.2 (varying from 1.66 to over 100) times higher than the probability of allocating a herd to a wrong level due to the test inaccuracy introduced by going from double to single testing. This is, however, an average; a herd with a true prevalence close to one of the level border cut-offs (40 and 70% weighted seroprevalence, respectively) would have a higher risk of being allocated to a wrong level than a herd with a true prevalence far from the level border cut-offs. The results are based on the current Danish sample sizes in the surveillance scheme, which implies that 60, 75 or 100 samples are taken annually in a herd, depending on its size. Other sample sizes would produce other results. PMID- 12900156 TI - The agreement in isolation of Salmonella enterica IIIb 61:k:1,5,(7) from rectal swabs, faecal samples and ileo-caecal lymph nodes from sheep. AB - The agreements between various culturing-method combinations and specimens to detect sheep naturally infected by Salmonella enterica IIIb 61:k:1,5,(7) were tested. Rectal swabs, faecal samples and ileo-caecal lymph nodes were collected from each individual. Rectal swabs called "group I" (n = 54) were cultured directly on selective media (selenite cysteine, SC). Rectal swabs called "group II" (n = 47) were pre-incubated in buffered peptone water. The four other combinations of culturing-method and specimen (one lymph node and three faecal) in each of the two groups were cultured directly on SC. The results from all the combinations of method and specimens were compared to the result from the rectal swab test by kappa and McNemar's chi-square. Kappas for the agreement between rectal swabs and faecal sample tests were only 0.4. The lymph-node test agreed even-more poorly, with mean kappa value -0.02. The McNemar's test revealed that the discordancies between the rectal-swab test and all of the other tests went in both directions but sometimes did show bias (directionality) in the discordancies. PMID- 12900157 TI - Dam-related effects on heart girth at birth, morbidity and growth rate from birth to 90 days of age in Swedish dairy calves. AB - We investigated the effects of dam-related factors (such as calving performance, milk leakage, diseases, milk production, and somatic-cell count (SCC)) on heart girth at birth and the incidence risk of diarrhoea and respiratory disease during the first 90 days in Swedish dairy calves. The effects of these dam-related factors and environmental and management-related (but not dietary) factors on the calves' growth rate during the first 90 days of life also were analysed. The study used nearly 3,000 heifer calves born in 1998 on 122 farms in the south-west of Sweden. Individual health records were kept by the farmers and visiting project veterinarians. The calf's heart girth was measured at birth and weaning. We used generalised linear mixed models for the size of the calf at birth and growth rate. Variables associated with the heart girth at birth were breed, calving performance, mastitis in the dam in the last 49 days before calving, milk production and parity. Variables associated with the growth rate were breed, calving performance, disease in the calf during its first 90 days of life, heart girth at birth, and housing of calves. The effect of the dam on the relative risk of diarrhoea and/or respiratory disease in the calf was evaluated by a generalised linear mixed model with a logit link. Morbidity in the dam during late pregnancy, retained placenta and SCC were associated with the relative risk of respiratory disease in the calf. None of the explanatory variables (other then breed) was associated with the relative risk of diarrhoea. PMID- 12900158 TI - Evaluation of herd sampling for Salmonella isolation on midwest and northeast US dairy farms. AB - Epidemiologic investigations of Salmonella infections in dairy cattle often rely on testing fecal samples from individual animals or samples from other farm sources to determine herd infection status. The objectives of this project were to evaluate the effect of sampling frequency on Salmonella isolation and to compare Salmonella isolation and serogroup classification among sample sources on 12 US dairy farms sampled weekly for 7-8 weeks. Three herds per state were enrolled from Michigan, Minnesota, New York and Wisconsin based upon predefined herd-size criteria. Weekly samples were obtained from cattle, bulk tank milk, milk filters, water and feed sources and environmental sites. Samples were submitted to a central laboratory for isolation of Salmonella using standard laboratory procedures. The herd average number of cattle fecal samples collected ranged from 26 to 58 per week. Salmonella was isolated from 9.3% of 4049 fecal samples collected from cattle and 12.9% of 811 samples from other sources. Serogroup C1 was found in more than half of the samples and multiple serogroups were identified among isolates from the same samples and farms. The percentage of herd visits with at least one Salmonella isolate from cattle fecal samples increased with overall herd prevalence of fecal shedding. Only the three herds with an average fecal shedding prevalence of more than 15% had over 85% of weekly visits with at least one positive fecal sample. The prevalence of fecal shedding from different groups of cattle varied widely among herds showing that herds with infected cattle may be classified incorrectly if only one age group is tested. Testing environmental sample sources was more efficient for identifying infected premises than using individual cattle fecal samples. PMID- 12900159 TI - Longitudinal serological responses to Salmonella enterica of growing pigs in a subclinically infected herd. AB - A longitudinal survey was conducted in France in a subclinically Salmonella infected farrow-to-finish pig farm to describe the time-course of the serological response to Salmonella enterica in growing pigs. We used three batches of sows and their corresponding litters (n = 31 litters). Among these, 256 pigs randomly selected and individually identified were followed from the first week of age until slaughter. Serial individual blood samples were submitted to indirect Salmonella-ELISA testing. Salmonella shedding was investigated by bacteriological testing of faecal material regularly collected from the sows and pigs and by environment swabs taken from the pens. Caecal contamination was checked at the slaughterhouse. Information about litter composition (filiation), location of the pigs in post-weaning and fattening pens, sanitary events, sex and body weights was recorded. 11.6% of the pigs shed S. enterica; 52% of pigs seroconverted before slaughter. The age-related variation of the natural logarithm of calibrated optical densities (COD) of pigs was described with two linear mixed models. From 8 to 61 days of age, the decrease in COD with age was fitted with a model including random effects of the animal and the dam on the intercept and slope, a batch random effect on the intercept and an individual birth-weight fixed effect on the intercept. The dam random effect was explained by the parity of the sow, Salmonella shedding by the sow during the farrowing phase and the value of the optical density of colostrum collected at parturition. A second model fitting the increase in COD from 61 days of age until slaughter included the random effect on intercept of the batch and the random effects on slope and intercept of the animal, the dam and the pen in which the followed animals were located during the fattening phase and the environmental contamination as fixed effect. In this second model, no relation was found between individual slaughter bacteriological results and increasing COD values. Considering seroconversion time between 61 days of age and slaughter, survival function were constructed using the Cox proportional-hazards model. Both methods suggested that seroconversions generally occurred during the last third of the fattening phase (from 140 days of age to slaughter), while shedding was observed during the first half of the fattening period. The fitted models suggest the existence of clusters (such as pen and litter of origin). PMID- 12900160 TI - Associations between leptospiral infection and seropositivity in rodents and environmental characteristics in Argentina. AB - Our objective was to look for associations between leptospiral infection in rodents and selected environmental and rodent characteristics in Santa Fe, Argentina. Rodents (n = 214) were trapped alive from January 1998 to December 1999 in three environmental settings. Kidneys from 118 rodents were cultured and serum samples from 201 were processed by enzyme-linked immunosorbent assay (ELISA). Logistic regression was performed with ELISA seropositivity as the dependent variable and rodent characteristics were offered as independent variables. Overall prevalence of positive ELISA reactions was 42% (84/201). In urban areas, leptospiral isolations belonged to the Ballum serogroup; in natural corridors, they belonged to the Icterohaemorragiae serogroup. M. musculus (house mouse) was the most-frequently captured species and the predominant one in urban areas. Most isolates and seropositivity results were obtained on this species. Adults and subadults had higher seroprevalences than juvenile rodents. Oligoryzomys flavescens had higher seroprevalence than Akodon azarae, Mus musculus, Rattus rattus and Rattus norvegicus. PMID- 12900161 TI - Herd-level ELISA seroprevalence of bovine viral diarrhea antibodies in bulk-tank milk in Chilean dairy herds. AB - Our objective in this cross-sectional study was to determine the seroprevalence in 1996 of commercial dairy herds in central Chile positive to bovine viral diarrhea (BVD) virus, by the detection of antibodies in bulk-tank milk samples. Samples were obtained from 501 dairy farms ( approximately 50,000 cows) from the Metropolitan, VI, VII and VIII regions of Chile. An indirect ELISA was used to detect BVD antibodies. An absorbance (A450) > or = 0.55 was considered positive. The apparent proportion of positive herds ranged from 71.2 to 83% by region and did not differ by region. We concluded that exposure to BVD was widely distributed in the central area of Chile in 1996. PMID- 12900162 TI - Evidence for a new field Culicoides vector of African horse sickness in South Africa. AB - Between February and May 1998, approximately 100 horses died of African horse sickness (AHS) in the cooler, mountainous, central region of South Africa. On 14 affected farms, 156,875 Culicoides of 27 species were captured. C. imicola Kieffer, hitherto considered the only field vector for AHS virus (AHSV), constituted <1% of the total Culicoides captured, and was not found on 29% of the farms. In contrast, 65% of the Culicoides were C. bolitinos Meiswinkel, and was found on all farms. Five isolations of AHSV were made from C. bolitinos, and none from 18 other species of Culicoides (including C. imicola). PMID- 12900163 TI - Seroprevalence of Johne's-disease infection in dairy cattle in California, USA. AB - A total of 1,950 serum samples from dairy cattle in California, USA were tested for the presence of antibodies to Mycobacterium avium subspecies paratuberculosis (MAP) using a commercial enzyme-linked immunosorbent assay (ELISA) kit. The sampled animals came from 65 herds and were sampled to reflect the relative numbers and distribution of dairy herds within three geographic regions in the state. Using the manufacturers suggested cut-off for a positive test, 89 animals (4.6%) were positive. The seroprevalence was 6.9% in the northern region of the state, 3.7% in the central region and 5.2% in the southern region. Using the sensitivity and specificity claimed by the manufacturer of the ELISA kit, the true prevalence in California dairy cattle overall was calculated as 9.4% (99% CI, 7.7%, 11.1%) and the true prevalences for the northern, central and southern regions were 14.1% (99% CI, 9.6%, 18.65%), 7.5% (99% CI, 5.6%, 9.4%), and 10.6% (99% CI, 5.9%, 15.6%), respectively. PMID- 12900164 TI - Neural connections that compute. AB - The UK's Foresight Cognitive Systems Project brings together researchers in the life sciences and physical sciences to see where they can learn from one another and to debate, and plan, the future of research in cognitive systems. The project, a part of the UK government's Foresight initiative, sets out to identify potential opportunities for the economy or society from new science and technology. Through a series of research reviews, the project has created 'snap shots' of research in cognitive systems. A major conference in Bristol in September will consider series of research manifestos created by interdisciplinary groups. PMID- 12900165 TI - Numb and Numblike control cell number during vertebrate neurogenesis. AB - A fundamental problem in nervous system development is how to control progenitor cell divisions to generate the correct number of cells for the mature brain and spinal cord. A recent report by Petersen and colleagues has identified an essential role for Numb and a related protein, Numblike, in this process. This study demonstrates that Numb and Numblike function to maintain the self-renewal properties of neural progenitor cells in the vertebrate neural tube. PMID- 12900166 TI - Does neurofilament phosphorylation regulate axonal transport? AB - Phosphorylation of neurofilaments has long been considered to regulate their axonal transport rate and, in doing so, to provide stability to mature axons. Interpretation of data recently obtained following C-terminal deletion experiments has prompted a challenge to this hypothesis. We present evidence that these deletion studies remain consistent with, rather than refute, a role for C terminal phosphorylation in regulation of neurofilament axonal transport. PMID- 12900167 TI - Persistent activity and memory in the entorhinal cortex. AB - The entorhinal cortex (EC) is an essential component of the medial temporal lobe long-term-memory system. Recently, Egorov et al. demonstrated that neurons in layer 5 of the EC show graded persistent activity: each neuron could maintain a constant rate firing over several minutes, even in the absence of input from other neurons. These findings suggest that layer-5 EC neurons could play a central role in the formation of associations among stimuli that occur at different times. PMID- 12900168 TI - Alcohol and error processing. AB - A recent study indicates that alcohol consumption reduces the amplitude of the error-related negativity (ERN), a negative deflection in the electroencephalogram associated with error commission. Here, we explore possible mechanisms underlying this result in the context of two recent theories about the neural system that produces the ERN - one based on principles of reinforcement learning and the other based on response conflict monitoring. PMID- 12900169 TI - Potential roles of insulin and IGF-1 in Alzheimer's disease. AB - Aging is characterized by a significant decline of metabolic and hormonal functions, which often facilitates the onset of severe age-associated pathologies. One outstanding example of this is the reported association of deranged signaling by insulin and insulin-like-growth-factor 1 (IGF-1) with Alzheimer's disease (AD). Recent compelling biological data reveal effects of insulin and IGF-1 on molecular and cellular mechanisms underlying the pathology of AD. This review discusses available biological data that highlight the therapeutic potential of the insulin-IGF-1 signaling pathway in AD. PMID- 12900170 TI - 'Unfolding' pathways in neurodegenerative disease. AB - The endoplasmic reticulum responds to stress by initiating a cascade of events known as the 'unfolded-protein response' (UPR). The accumulation of misfolded proteins in the leukodystrophy Pelizaeus-Merzbacher disease activates this stress response, resulting in apoptosis of oligodendrocytes. Although it remains uncertain whether the UPR plays a mechanistic role in prototypical neurodegenerative disorders such as Alzheimer's disease, this is plausible because misfolded proteins are directly implicated in the pathogenesis of these disorders. PMID- 12900171 TI - Oligodendrocytes, GRPs and MNOPs. PMID- 12900172 TI - Synaptotagmin I, a Ca2+ sensor for neurotransmitter release. AB - The discovery that Ca(2+) triggers rapid neurotransmitter release has prompted the search for the Ca(2+) sensor. There is now general agreement that the vesicle associated Ca(2+)-binding protein, synaptotagmin I, is required for the tight temporal coupling between Ca(2+) influx and synaptic vesicle fusion. However, the precise mechanism of Ca(2+)-sensing by synaptotagmin I is still under debate despite intensive investigation using genetic, biochemical and electrophysiological tools. Here, we discuss many of the genetic manipulations from the past few years that have shed light on the Ca(2+)-sensing function of synaptotagmin I. We also present our view as to how the Ca(2+) signal is translated rapidly into membrane fusion at fast chemical synapses. PMID- 12900173 TI - A computational substrate for incentive salience. AB - Theories of dopamine function are at a crossroads. Computational models derived from single-unit recordings capture changes in dopaminergic neuron firing rate as a prediction error signal. These models employ the prediction error signal in two roles: learning to predict future rewarding events and biasing action choice. Conversely, pharmacological inhibition or lesion of dopaminergic neuron function diminishes the ability of an animal to motivate behaviors directed at acquiring rewards. These lesion experiments have raised the possibility that dopamine release encodes a measure of the incentive value of a contemplated behavioral act. The most complete psychological idea that captures this notion frames the dopamine signal as carrying 'incentive salience'. On the surface, these two competing accounts of dopamine function seem incommensurate. To the contrary, we demonstrate that both of these functions can be captured in a single computational model of the involvement of dopamine in reward prediction for the purpose of reward seeking. PMID- 12900174 TI - Hemispheric asymmetries in cerebral cortical networks. AB - Since the middle of the 19th century it has been recognized that several higher cognitive functions, including language, are lateralized in cerebral cortex. Neuropsychological studies on patients with brain lesions and rapid developments in brain imaging techniques have provided us with an increasing body of data on the functional aspects of language lateralization, but little is known about the substrate on which these specializations are realized. Much attention has been focused on the gross size and shape of cortical regions involved, but recent findings indicate that the columnar and connectional structure within auditory and language cortex in the left hemisphere are distinct from those in homotopic regions in the right hemisphere. These findings concern parameters that are closely linked to the processing architecture within the respective regions. Thus, the comparison of these microanatomical specializations with their respective functional counterparts provides important insights into the functional role of cerebral cortical organization and its consequences for processing of cortical information in the implementation of complex cognitive functions. PMID- 12900175 TI - When inhibition goes incognito: feedback interaction between spiny projection neurons in striatal function. AB - Theories about basal ganglia function have always been driven by our knowledge about the spiny projection neurons of the striatum. At the core of these theories lies the question of how, precisely, spiny projection neurons process cortical inputs. Most recently, studies demonstrating the role of spiny projection neurons in local synaptic GABA transmission have provided several new avenues for exploring striatal dynamics. They have also suggested new experimental directives for examining the specific ways in which spiny projection neurons both compete and cooperate through their local axon collaterals during cortical input processing. PMID- 12900176 TI - Visualizing phosphoinositide signalling in single neurons gets a green light. AB - There is now substantial evidence, from single-cell imaging, that complex patterns of release from Ca(2+) stores play an important role in regulating synaptic efficacy and plasticity. Moreover, the major mechanism of store release depends on the generation of inositol 1,4,5-trisphosphate [Ins(1,4,5)P(3)] through the action of phospholipase(s) C on phosphatidylinositol 4,5-bisphosphate [PtdIns(4,5)P(2)], and several neurotransmitters can enhance receptor-mediated activation of this enzyme. The recent development of techniques to image real time changes in PtdIns(4,5)P(2) hydrolysis according to generation of Ins(1,4,5)P(3) and diacylglycerol in single cells has significantly advanced our ability to investigate these signalling pathways, particularly in relation to single-cell Ca(2+) signals. This article reviews these new approaches and how they have provided novel insights into mechanisms underlying spatio-temporal Ca(2+) signals and phospholipase C activation in neurons. PMID- 12900177 TI - Cerebrovascular modelling: a union of physiology, clinical medicine and biomedical engineering. PMID- 12900178 TI - The critical closing pressure of the cerebral circulation. AB - The critical closing pressure (CrCP) of the cerebral circulation indicates the value of arterial blood pressure (ABP) at which cerebral blood flow (CBF) approaches zero. Measurements in animals and in humans, have shown that the CrCP is significantly greater than zero. A simple mathematical model, incorporating the effects of arterial elasticity and active wall tension, shows that CrCP can be influenced by several structural and physiological parameters, notably intracranial pressure (ICP) and active wall tension. Due to the non-linear shape of the complete ABP-CBF curve, most methods proposed for estimation of CrCP can only represent the linear range of the pressure-flow (or velocity) relationship. As a consequence, only estimates of apparent CrCP can be obtained, and these tend to be significantly higher than the true CrCP. Estimates of apparent CrCP have been shown to be influenced by arterial PCO2, ICP, cerebral autoregulation, intra thoracic pressure, and mean ABP. There is a lack of investigation, under well controlled conditions, to assess whether CrCP is altered in disease states. Studies of the cerebral circulation need to take CrCP into account, to obtain more accurate estimates of cerebrovascular resistance changes, and to reflect the correct dynamic relationship between instantaneous ABP and CBF. PMID- 12900179 TI - Linearity and non-linearity in cerebral hemodynamics. AB - BACKGROUND: Transcranial Doppler ultrasound has been extensively used to study cerebral hemodynamics, and yet the basic characteristics of the input/output system of blood pressure/velocity are little known. We examine whether this system can best be considered linear or non-linear. METHODS: We assessed the adequacy of linear modeling in four ways: (1) Known properties of cerebral blood flow were reviewed and analyzed from a systems standpoint; (2) 1100 ARX & OE model types were tested with data from 29 normal subjects, with and without lowpass filtering; (3) time-frequency analysis was used to identify nonstationary behavior and markers of non-linearity (such as bifurcations, chirps, and intermittent autoregulatory impairment) in the same data sets; (4) simple computer models of autoregulation incorporating time delays and non-linear elements were tested for production of spontaneous oscillations. RESULTS: (1) Several aspects of cerebral hemodynamics are poorly described by linear models, (2) the ARX & OE models performed poorly, (3) time-frequency analysis showed non linear and nonstationary behavior, (4) the computer models produced spontaneous oscillations similar to those observed in humans. CONCLUSIONS: There is strong evidence that the blood pressure/velocity system is non-linear. PMID- 12900180 TI - Dynamic cerebral autoregulation assessment using an ARX model: comparative study using step response and phase shift analysis. AB - Middle cerebral arterial blood velocity (MCAv) response to spontaneous and manipulated changes of arterial blood pressure (ABP) was studied in eight subjects using a linear autoregressive with exogenous input (ARX) model. ABP and MCAv were measured non-invasively by photoplethysmograph and transcranial Doppler ultrasound, respectively. Data were recorded at rest (spontaneous changes in ABP) and during thigh cuff (step-wise changes) and lower body negative pressure (sinusoidal changes of 1/12 Hz) tests in both normocapnia and hypercapnia (5% CO2). Since autoregulation is modulated by CO2, respiratory CO2 was simultaneously monitored to allow comparison of cerebral autoregulation status with different CO2 levels. ABP and MCAv were fitted by ARX models and dynamic cerebral autoregulation was estimated by analysing both the step responses and phase shift at the 1/12 Hz of the corresponding ARX models. The ARX model consistently modelled the phase lead of MCAv to ABP and it showed that the phase shift at 1/12 Hz of ARX model is consistent with the real phase shift of the data (p=0.59). Strong linear relationships between pCO2 and gradient of the step response (r=-0.58, p<0.0001) and between pCO2 and phase shift (r=-0.76, p<0.0001) were observed, which suggests that cerebral autoregulation can be assessed by step response or phase shift analysis of the ARX model fitted to ABP and MCAv data with spontaneous changes. PMID- 12900181 TI - Quantitative assessment of cerebral autoregulation from transcranial Doppler pulsatility: a computer simulation study. AB - Transcranial Doppler (TCD) ultrasonography is largely used today to achieve non invasive assessment of cerebral autoregulation and cerebrovascular reactivity in neurosurgical patients. Recent experimental and clinical studies suggest that not only the pattern of mean velocity, but also velocity pulse amplitude alterations during changes in cerebral perfusion pressure (CPP) contain information on autoregulation status. The aim of this work is to investigate the relationship between cerebral autoregulation and TCD pulsatility by means of a comprehensive mathematical model of intracranial dynamics and cerebrovascular regulation. Simulation results, performed using different values of the most important clinical parameters of the model (autoregulation strength, cerebrospinal fluid (CSF) outflow resistance and intracranial elastance coefficient) show that velocity pulse amplitude increases with a reduction in CPP in patients with intact autoregulation, whereas changes in velocity pulsatility are modest in patients with weak autoregulation. Finally, velocity pulse amplitude decreases during a CPP reduction in patients with impaired autoregulation. Moreover, the relationship between the velocity pulse amplitude changes and autoregulation strength is almost linear in a wide range of CPP values, and is scarcely affected by changes in CSF circulation and intracranial elasticity. Starting from these results, we suggest a new quantitative index to assess autoregulation strength, i.e. G(aut)% = (s-b)/a, where G(aut)% is autoregulation strength (100% means intact autoregulation, 0% means impaired autoregulation), a approximately -0.03; b approximately 1.5 and s is the slope of the relationship ' percentage changes of velocity pulse amplitude to arterial pressure pulse amplitude vs. CPP changes'. PMID- 12900182 TI - Implementation of non-invasive brain physiological monitoring concepts. AB - The paper presents innovative methods and technology for non-invasive intracranial hemodynamics monitoring based on the measurement of brain parenchyma acoustic properties. The clinical investigation of new technology shows the similarity between the invasively recorded intracranial pressure (ICP) and non invasively recorded intracranial blood volume (IBV) pulse waves, slow waves and slow trends under intensive care unit (ICU) conditions. Also, the applicability of the non-invasive IBV slow wave monitoring technique for cerebrovascular autoregulation non-invasive long-term monitoring is demonstrated by theoretical and experimental studies. PMID- 12900183 TI - Intracranial pressure dynamics: changes of bandwidth as an indicator of cerebrovascular tension. AB - The transmission bandwidth (BW) of arterial blood pressure (ABP) to intracranial pressure (ICP) was examined as a means of bedside monitoring of the state of cerebrovascular tension. Changes of BW of a black box identification model, relative arteriolar resistance and intracranial compliance were obtained from a piglet model equipped with a cranial window during induction of asphyxia, hypercapnia, and hypoxia. Changes of black box BW values and simulated changes of BW produced by a physiologically based lump parameter model of ICP dynamics are used to evaluate the hypothesis that during active cerebrovascular tension, changes of BW are inversely related to cerebral perfusion pressure (CPP), and during passive cerebrovascular tension, changes of BW are not inversely related to changes of CPP. Induction of asphyxia (n = 3) produced BW changes of the black box model that were simulated as an active cerebrovascular tension phase during decreasing CPP followed by a passive tension phase. Reventilation after prolonged asphyxia produced significant increases of BW that were simulated by a passive tension. Hypercapnic (n = 6) and hypoxic (n = 6) challenges produced: (1) significant changes of BW that were matched with simulations of the lumped parameter model for active tension; and (2) relationships between values of BW and relative average cerebral arteriolar resistance and intracranial compliance were inverse and correlated to a regression function of approximately x(-1). Changes of BW of the black box model and the simulations of the lumped parameter model support the feasibility of the stated hypothesis. As such, the evaluation of changes of BW of the black box model with respect to changes of CPP may be a useful method for monitoring the state of cerebrovascular tension. PMID- 12900184 TI - The myogenic response in isolated rat cerebrovascular arteries: smooth muscle cell model. AB - Previous models of the cerebrovascular smooth muscle cell have not addressed the interaction between the electrical, chemical, and mechanical components of cell function during the development of active tension. These models are primarily electrical, biochemical or mechanical in their orientation, and do not permit a full exploration of how the smooth muscle responds to electrical or mechanical forcing. To address this issue, we have developed a new model that consists of two major components: electrochemical and chemomechanical subsystem models of the smooth muscle cell. Included in the electrochemical model are models of the electrophysiological behavior of the cell membrane, fluid compartments, Ca2+ release and uptake by the sarcoplasmic reticulum (SR), and cytosolic Ca2+ buffering, particularly by calmodulin (CM). With this subsystem model, we can study the mechanics of the production of intracellular Ca2+ transient in response to membrane voltage clamp pulses. The chemomechanical model includes models of: (a) the chemical kinetics of myosin phosphorylation, and the formation of phosphorylated (cycling) myosin cross-bridges with actin, as well as attached (non-cycling) latch-type cross-bridges; and (b) a model of force generation and mechanical coupling to the contractile filaments and their attachments to protein structures and the skeletal framework of the cell. The two subsystem models are tested independently and compared with data. Likewise, the complete (combined) cell model responses to voltage pulse stimulation under isometric and isotonic conditions are calculated and compared with measured single cell length-force and force-velocity data obtained from literature. This integrated cell model provides biophysically based explanations of electrical, chemical, and mechanical phenomena in cerebrovascular smooth muscle, and has considerable utility as an adjunct to laboratory research and experimental design. PMID- 12900185 TI - The myogenic response in isolated rat cerebrovascular arteries: vessel model. AB - We develop an integrated model of isolated rat arterial resistance vessel (RV), which can simulate its major property of myogenic response. The vascular smooth muscle cell is an important component of the wall of this vessel, and serves as a vasomotor organ providing the active tension generation that underlies the myogenic response of the wall to stretch. In the previous study, we focused on the development of a smooth muscle cell model that can mimic the strain-sensing and force-generating features of the myogenic mechanism. In the current model, we embed this cell model in a larger vessel wall configuration, and couple the time course of cellular contractile activation to macroscopic changes in vessel diameter. The integrated model is used to mimic published pressure-vessel diameter data obtained from isolated RVs that are mounted in a hydraulic test apparatus. The model provides biophysically based insights into the myogenic mechanism as it responds to changes in transmural pressure, in the presence and absence of Ca2+ blockers applied to the bathing fluid.It mimics measured data very well and provides a model that is able to link events at subcellular level to macroscopic changes in vessel diameter. The model initiates a mechanistic approach to investigate myogenic response, which has not been taken previously by any other models. PMID- 12900186 TI - Learning from others: overseas nurses' views of UK nursing. PMID- 12900187 TI - International nursing students: a phenomenological perspective. AB - Colleges and universities are faced with the challenge of how to create learning environments relevant to the internationalization of society and culture. International learning experiences, the focus of this article, may be one effective tool for internationalizing nursing curricula. Baccalaureate nursing students with international education experiences who were residing in a rural USA state were invited to participate in a qualitative descriptive study informed by phenomenology. Sixteen nursing students participated in mail surveys and eleven in interviews. Participants were mainly female, Caucasian, university seniors with an average age of 31 years. Thematic statements were isolated into nine experiential structures and four essential themes. Themes were identified as preparing, adjusting, caring, and transforming and were found to be influenced by culture, values, and ethics. A descriptive diagram was created summarizing the structure of meaning. Significance of this nursing study was the identification of plausible descriptive insights that may be useful to educators who assist students with international education experiences. PMID- 12900188 TI - Cross-infection prevention, basic hygiene practices and education within nursing and health care in Latvia: a Swedish-Latvian practice development project. AB - The primary aim of this practice development project was to explore the level of knowledge of nursing and medical staff at the Infection Clinic at Liepaja regarding the spread of infectious diseases. Arrangements were also made for some of the Latvian health care staff to visit the Infection Clinic in Gothenburg to increase their knowledge about basic hygiene and to enable them to study the research in this area. Later we were able to formulate this new knowledge into written guidelines. The theoretical education started with questions regarding staffs expertise in preventing the spread of infections. Areas covered were handwashing, hand disinfection, use of disposable gloves and protective clothing. This revealed a need for development of both theoretical and practical knowledge in this area. However, hygiene practices improved after the theoretical education and the visits to the Infection Clinic in Gothenburg. On our return visit to Liepaja dispensers of liquid soap together with hand disinfectants were evident at every wash basin on the unit. Disposable gloves were also routinely used for the dressing of wounds and invasive procedures. Furthermore, disposable coats and masks were used when caring for highly infectious patients. The key cultural differences were the lack of nursing documentation and the relative absence of a dialogue directly between the nurse and the patient. PMID- 12900189 TI - An educational panopticon? New technology, nurse education and surveillance. AB - This papers raises concerns about the use of new technology in nurse education. It is possible that new forms of computer-based learning and teaching carry with them the possibility of new kinds of 'panoptic' surveillance and control of students. We discuss how this is possible, and set this development within a wider social context. These forms of panoptic surveillance are, we believe, inimical to the underlying values and philosophy of nurse education. New technology does not necessarily have to be used in this way, and can be part of a liberating pedagogy. We would not propose that surveillance of students should never be used, but rather that its use should be considered and debated. PMID- 12900190 TI - Nursing students' and lecturers' perspectives of objective structured clinical examination incorporating simulation. AB - Objective Structured Clinical Examination (OSCE) has been widely and increasingly used since it was developed. Research has shown that it is an effective evaluation tool to assess practical skills (Sloan et al. 1995). In many instances the OSCE process has been adapted to test trainees from different healthcare related disciplines. In nursing education, as presented in this paper, principles of OSCE can also be used in a formative way to enhance skill acquisition through simulation. The aim of this approach to teaching is safely to help students gain more confidence when confronted by technical instruments present in the hospital environment, and to encourage them to reflect on a range of skills and competences they need to acquire. The OSCE stations can be designed in the form of small scenarios where students have to set-up or interact with technical instruments, or communicate with patients. This type of simulation exercise can be varied as a whole and specifically within each station at the same time. The use of this hybrid formative OSCE is being assessed by nursing students and lecturers. The feedback received regarding this teaching method and the results of this study are useful and show that OSCE are favourably perceived. PMID- 12900191 TI - Strategies for interprofessional education: the Interprofessional Team Objective Structured Clinical Examination for midwifery and medical students. AB - This paper provides an overview of the processes involved in implementing an interprofessional education (IPE) strategy in a recently established School of Human Development at the University of Nottingham. The merger of the academic divisions of child health, midwifery, obstetrics and gynaecology was a deliberate initiative to create an organisational infrastructure intended to enhance opportunities for interprofessional collaborations in research and education. As a first step, a small group of academic midwives and obstetricians formed a project group to find the best way of facilitating IPE for medical and midwifery students at undergraduate level. A discussion is provided of the work the project group undertook to: determine an agreed definition of IPE; decide an action research approach was needed; determine the ways in which teaching and learning strategies were to be implemented, evaluated and compared; and identify the factors inhibiting and enhancing developments. Evaluations have demonstrated that the Interprofessional Team Objective Structured Clinical Examination (ITOSCE) focusing on intrapartum scenarios is effective in promoting interprofessional learning. Both medical and midwifery students and facilitators agree that team working and understanding each other's roles has been enhanced and that although resource intensive, IPE is worth the time and effort involved. PMID- 12900192 TI - Australian nurses' perception of the impact of their postgraduate studies on their patient care activities. AB - Postgraduate nursing education, formal or informal, has grown enormously and the benefits, to the students, their employers or their patients or its impact on clinical practice has not been well researched, particularly in Australia. The authors commenced a 10 year longitudinal study at a university in Sydney, Australia to track five cohorts of postgraduates at two yearly intervals to determine their career paths, changes in professional behaviour and the perceived impact of their postgraduate education on the care they deliver. This paper will report these five cohorts' (N=236) perceptions of the impact of their postgraduate studies on their care delivery two years after completion of their study. The results indicate that their postgraduate education had a positive to strongly positive impact on the majority of items. These as well as those items identified as not being affected will also be presented and discussed. PMID- 12900193 TI - Theatre and pedagogy: using drama in mental health nurse education. AB - Drama is a feature of all civilisations. It has long been used as an educative tool. In this paper, the authors advocate for the use of drama in mental health nurse education. An historical overview of the relationship between drama and education is offered. Through their own experiences the authors give examples of the use of drama in the classroom. Drama challenges the power position of the participants, teacher and student alike. A humanistic framework is offered that promotes empathic understanding between all those concerned, especially the client. PMID- 12900194 TI - Why do student nurses leave? Suggestions from a Delphi study. AB - The high dropout rate of nursing students is a major concern. However, there is little research available about the reasons why students leave. Universities collect some information from 'exit' interviews but, due to ethical sensitivities, it is not made available for research analysis. The purpose of this study was to establish a consensus view of the reasons why student nurses leave their pre-registration education programme. The study was undertaken in two phases. Initially, an exploratory phase using focus groups and one-to-one interviews was used to gather multi-professional views about the reasons why students leave. In the second phase a questionnaire was developed from the themes arising from the data analysis in phase one. The questionnaire was administered to an expert panel of student nurses in the form of a three-round Delphi Study. The consensus level was set at 75%. It is important to clarify that, for reasons stated above, the views expressed in this paper are those of current students about others who had left the programme. From this study, it is apparent that, with the exception of academic failure, there was no single contributing factor that was thought to make students leave. However, there were a number of important issues identified as factors that may result in student nurses leaving. These include communication and operational factors between the university and clinical areas, feelings of not being valued, unmet expectations, and stress. These issues were of concern to students and appeared to have a cumulative effect that led them to question whether they should continue their education programme. On the basis of these findings, several recommendations are made to improve the student nurse experience. PMID- 12900195 TI - Pharmacology education: a theoretical framework of applied pharmacology and therapeutics. AB - United Kingdom Government initiatives such as The NHS Plan (DoH 2000) identified the need for nurses to undertake multi-skilled professional roles. In the United Kingdom, the transition to a larger graduate nurse workforce continues, as Universities offer nurse pre-registration courses at diploma and degree levels. Concomitant with the change in educational standard is the need to teach student nurses the theoretical principles of medication management and to develop skills in clinical reasoning skills. Both elements are limited in current educational pre-registration nurse programmes, in order to develop the future, multi-skilled workforce such courses should incorporate the theory and skills of health assessment, physical examination, applied pharmacology and clinical reasoning. This paper aims to examine how knowledge of applied pharmacology and therapeutics can be integrated into an undergraduate pre-registration nursing programme. Discussion focuses on how this generic framework can provide educators with an outline of the theoretical constructs, their application, the teaching strategies involved and instruction on how to prepare nurses to clinically reason with regard to medication management issues. This framework can be adapted to accommodate nurses studying for all parts of the register. PMID- 12900201 TI - Physical theory, origin of flight, and a synthesis proposed for birds. AB - Neither flapping and running to take-off nor gliding from heights can be disproved as the assured evolutionary origin of self-powered flight observed in modern vertebrates. Gliding with set wings would utilize available potential energy from gravity but gain little from flapping. Bipedal running, important in avian phylogeny, possibly facilitated the evolution of flight. Based on physical principles, gliding is a better process for the origin of powered flight than the "ground-up" process, which physically is not feasible in space or time (considering air resistance, metabolic energy costs, and mechanical resistance to bipedal running). Proto-avian ancestors of Archaeopteryx and Microraptor probably flapped their sparsely feathered limbs synchronously while descending from leaps or heights, with such "flutter-gliding" presented as a synthesis of the two earlier theories of flight origin (making use of the available potential energy from gravity, involving wing thrusts and flapping, coping with air resistance that slows air speed, but effecting positive fitness value in providing lift and slowing dangerous falls). PMID- 12900200 TI - SARS outbreaks in Ontario, Hong Kong and Singapore: the role of diagnosis and isolation as a control mechanism. AB - In this article we use global and regional data from the SARS epidemic in conjunction with a model of susceptible, exposed, infective, diagnosed, and recovered classes of people ("SEIJR") to extract average properties and rate constants for those populations. The model is fitted to data from the Ontario (Toronto) in Canada, Hong Kong in China and Singapore outbreaks and predictions are made based on various assumptions and observations, including the current effect of isolating individuals diagnosed with SARS. The epidemic dynamics for Hong Kong and Singapore appear to be different from the dynamics in Toronto, Ontario. Toronto shows a very rapid increase in the number of cases between March 31st and April 6th, followed by a significant slowing in the number of new cases. We explain this as the result of an increase in the diagnostic rate and in the effectiveness of patient isolation after March 26th. Our best estimates are consistent with SARS eventually being contained in Toronto, although the time of containment is sensitive to the parameters in our model. It is shown that despite the empirically modeled heterogeneity in transmission, SARS' average reproductive number is 1.2, a value quite similar to that computed for some strains of influenza (J. Math. Biol. 27 (1989) 233). Although it would not be surprising to see levels of SARS infection higher than 10% in some regions of the world (if unchecked), lack of data and the observed heterogeneity and sensitivity of parameters prevent us from predicting the long-term impact of SARS. The possibility that 10 or more percent of the world population at risk could eventually be infected with the virus in conjunction with a mortality rate of 3 7% or more, and indications of significant improvement in Toronto support the stringent measures that have been taken to isolate diagnosed cases. PMID- 12900202 TI - Sperm competition games: sperm selection by females. AB - We analyse a co-evolutionary sexual conflict game, in which males compete for fertilizations (sperm competition) and females operate sperm selection against unfavourable ejaculates (cryptic female choice). For simplicity, each female mates with two males per reproductive event, and the competing ejaculates are of two types, favourable (having high viability or success) or unfavourable (where progeny are less successful). Over evolutionary time, females can increase their level of sperm selection (measured as the proportion of unfavourable sperm eliminated) by paying a fecundity cost. Males can regulate sperm allocations depending on whether they will be favoured or disfavoured, but increasing sperm allocation reduces their mating rate. The resolution of this game depends on whether males are equal, or unequal. Males could be equal: each is favoured with probability, p, reflecting the proportion of females in the population that favour his ejaculate (the 'random-roles' model); different males are favoured by different sets of females. Alternatively, males could be unequal: given males are perceived consistently by all females as two distinct types, favoured and disfavoured, where p is now the frequency of the favoured male type in the population (the 'constant-types' model). In both cases, the evolutionarily stable strategy (ESS) is for females initially to increase sperm selection from zero as the viability of offspring from unfavourable ejaculates falls below that of favourable ejaculates. But in the random-roles model, sperm selection decreases again towards zero as the unfavourable ejaculates become disastrous (i.e. as their progeny viability decreases towards zero). This occurs because males avoid expenditure in unfavourable matings, to conserve sperm for matings in the favoured role where their offspring have high viability, thus allowing females to relax sperm selection. If sperm selection is costly to females, ESS sperm selection is high across a region of intermediate viabilities. If it is uncostly, there is no ESS in this region unless sperm limitation (i.e. some eggs fail to be fertilized because sperm numbers are too low) is included into the model. In the constant-types model, no relaxation of sperm selection occurs at very low viabilities of disfavoured male progeny. If sperm selection is sufficiently costly, ESS sperm selection increases as progeny viability decreases down towards zero; but if it is uncostly, there is no ESS at the lowest viabilities, and unlike the random-roles model, this cannot be stabilized by including sperm limitation. Sperm allocations in the ESS regions differ between the two models. With random roles, males always allocate more sperm in the favoured role. With constant types, the male type that is favoured allocates less sperm than the disfavoured type. These results suggests that empiricists studying cryptic female choice and sperm allocation patterns need to determine whether sperm selection is applied differently, or consistently, on given males by different females in the same population. PMID- 12900203 TI - The dynamics of gas bubbles in conduits of vascular plants and implications for embolism repair. AB - Pressure-induced tensions in the xylem, the water conducting tissue of vascular plants, can lead to embolism in the water-conducting cells. The details and mechanisms of embolism repair in vascular plants are still not well understood. In particular, experimental results which indicate that embolism repair may occur during xylem tension cause great problems with respect to current paradigms of plant water transport. The present paper deals with a theoretical analysis of interfacial effects at the pits (pores in the conduit walls), because it was suggested that gas-water interfaces at the pit pores may be involved in the repair process by hydraulically isolating the embolized conduit. The temporal behaviour of bubbles at the pit pores was especially studied since the question of whether these pit bubbles are able to persist is of crucial importance for the suggested mechanism to work. The results indicate that (1) the physical preconditions which are necessary for the suggested mechanism appear to be satisfied, (2) pit bubbles can achieve temporal stability and therefore persist and (3) dissolving of bubbles in the conduit lumen may lead to the final breakdown of the hydraulic isolation. The whole process is, however, complex and strongly dependent on the detailed anatomy of the pit and the contact angle. PMID- 12900204 TI - Simulation of circadian rhythm generation in the suprachiasmatic nucleus with locally coupled self-sustained oscillators. AB - In mammals, circadian rhythms are driven by a pacemaker located in the suprachiasmatic nuclei (SCN) of the anterior hypothalamus. The firing rate of neurons within the SCN exhibits a circadian rhythm. There is evidence that individual neurons within the SCN act as circadian oscillators. Rhythm generation in the SCN was therefore modeled by a system of self-sustained oscillators. The model is composed of up to 10000 oscillatory elements arranged in a square array. Each oscillator has its own (randomly determined) intrinsic period reflecting the widely dispersed periods observed in the SCN. The model behavior was investigated mainly in the absence of synchronizing zeitgebers. Due to local coupling the oscillators synchronized and an overall rhythm emerged. This indicates that a locally coupled system is capable of integrating the output of individual clock cells with widely dispersed periods. The period of the global output (average of all oscillators) corresponded to the average of the intrinsic periods and was stable even for small amplitudes and during transients. Noise, reflecting biological fluctuations at the cellular level, distorted the global rhythm in small arrays. The period of the rhythm could be stabilized by increasing the array size, which thus increased the robustness against noise. Since different regions of the SCN have separate output pathways, the array of oscillators was subdivided into four quadrants. Sudden deviations of periodicity sometimes appeared in one quadrant, while the periods of the other quadrants were largely unaffected. This result could represent a model for splitting, which has been observed in animal experiments. In summary, the multi-oscillator model of the SCN showed a broad repertoire of dynamic patterns, revealed a stable period (even during transients) with robustness against noise, and was able to account for such a complex physiological behavior as splitting. PMID- 12900205 TI - The kinetics of monospermic and polyspermic fertilization in free-spawning marine invertebrates. AB - The equation of Vogel et al. (1982) is widely used in fertilization studies of free-spawning marine invertebrates to predict the percentage of viable eggs that will be fertilized at any specified levels of gamete concentration and contact time. Here, the random collision model that underlies the Vogel et al. equation is extended to distinguish between monospermic and polyspermic fertilization, and separate equations for the percentages of monospermic and polyspermic fertilization are obtained. These equations provide an explanation for empirical observations which have shown a decreased percentage of successful egg development at high sperm concentrations. Comparison is made with an earlier heuristic attempt (Styan, 1998) to predict the extent of polyspermic fertilization, and it is found that this earlier method can underestimate the percentage of polyspermic fertilization by up to 10 percent. Moreover, the approach used here retains the flexibility to model changes in sperm concentration due to dispersal mechanisms, and is able to model different mechanisms for the block to polyspermy. PMID- 12900206 TI - Systemic metabolic reactions are obtained by singular value decomposition of genome-scale stoichiometric matrices. AB - Genome-scale metabolic networks can be reconstructed. The systemic biochemical properties of these networks can now be studied. Here, genome-scale reconstructed metabolic networks were analysed using singular value decomposition (SVD). All the individual biochemical conversions contained in a reconstructed metabolic network are described by a stoichiometric matrix (S). SVD of S led to the definition of the underlying modes that characterize the overall biochemical conversions that take place in a network and rank-ordered their importance. The modes were shown to correspond to systemic biochemical reactions and they could be used to identify the groups and clusters of individual biochemical reactions that drive them. Comparative analysis of the Escherichia coli, Haemophilus influenzae, and Helicobacter pylori genome-scale metabolic networks showed that the four dominant modes in all three networks correspond to: (1) the conversion of ATP to ADP, (2) redox metabolism of NADP, (3) proton-motive force, and (4) inorganic phosphate metabolism. The sets of individual metabolic reactions deriving these systemic conversions, however, differed among the three organisms. Thus, we can now define systemic metabolic reactions, or eigen-reactions, for the study of systems biology of metabolism and have a basis for comparing the overall properties of genome-specific metabolic networks. PMID- 12900207 TI - Exact solutions to the continuous-quality equation for soil organic matter turnover. AB - All living systems depend on transformations of elements between different states. In particular, the transformation of dead organic matter in the soil (SOM) by decomposers (microbes) releases elements incorporated in SOM and makes the elements available anew to plants. A major problem in analysing and describing this process is that SOM, as the result of the decomposer activity, is a mixture of a very large number of molecules with widely differing chemical and physical properties. The continuous-quality equation (CQE) is a general equation describing this complexity by assigning a continuous-quality variable to each carbon atom in SOM. The use of CQE has been impeded by its complicated mathematics. Here, we show by deriving exact solutions that, at least for some specific cases, there exist solutions to CQE. These exact solutions show that previous approximations have overestimated the rate by which litter decomposes and as a consequence underestimated steady state SOM amounts. The exact and approximate solutions also differ with respect to the parameter space in which they yield finite steady-state SOM amounts. The latter point is important because temperature is one of the parameters and climatic change may move the solution from a region of the parameter space with infinite steady-state SOM to a region of finite steady-state SOM, with potentially large changes in soil carbon stores. We also show that the solution satisfies the Chapman-Kolmogorov theorem. The importance of this is that it provides efficient algorithms for numerical solutions. PMID- 12900208 TI - Measuring mast seeding behavior: relationships among population variation, individual variation and synchrony. AB - Mast seeding, or masting, is the variable production of flowers, seeds, or fruit across years more or less synchronously by individuals within a population. A critical issue is the extent to which temporal variation in seed production over a collection of individuals can be viewed as arising from a combination of individual variation and synchrony among individuals. Studies of masting typically quantify such variation in terms of the coefficient of variation (CV). In this paper we examine mathematically how the population CV relates to the mean individual CV and synchrony, concluding that the relationship is a complex one which cannot isolate an overall measure of synchrony, and involves additional factors, principally the number of plants sampled and the mean productivity per plant. Our development suggests some simple approximate relationships of population CV to individual variability, synchrony and the number of individuals. These were found to fit quite well when applied to data from 59 studies which included seed production at the individual level. PMID- 12900209 TI - A tale of two defectors: the importance of standing for evolution of indirect reciprocity. AB - Indirect reciprocity occurs when the cooperative behavior between two individuals is contingent on their previous behavior toward others. Previous theoretical analysis indicates that indirect reciprocity can evolve if individuals use an image-scoring strategy. In this paper, we show that, when errors are added, indirect reciprocity cannot be based on an image-scoring strategy. However, if individuals use a standing strategy, then cooperation through indirect reciprocity is evolutionarily stable. These two strategies differ with respect to the information to which they attend. While image-scoring strategies only need attend to the actions of others, standing strategies also require information about intent. We speculate that this difference may shed light on the evolvability of indirect reciprocity. Additionally, we show that systems of indirect reciprocity are highly sensitive to the availability of information. Finally, we present a model which shows that if indirect reciprocity were to evolve, selection should also favor trusting behavior in relations between strangers. PMID- 12900210 TI - Microsatellite mutations during the polymerase chain reaction: mean field approximations and their applications. AB - We develop a novel mathematical model for microsatellite mutations during polymerase chain reaction (PCR). Based on the model, we study the first- and second-order moments of the number of repeat units in a randomly chosen molecule after n PCR cycles and their corresponding mean field approximations. We give upper bounds for the approximation errors and show that the approximation errors are small when the mutation rate is low. Based on the theoretical results, we develop a moment estimation method to estimate the mutation rate per-repeat-unit per PCR cycle and the probability of expansion when mutations occur. Simulation studies show that the moment estimation method can accurately recover the true mutation rate and probability of expansion. Finally, the method is applied to experimental data from single-molecule PCR experiments. PMID- 12900211 TI - Retroviral gene therapy--where now? PMID- 12900212 TI - Astrocytes: a cellular player in Abeta clearance and degradation. PMID- 12900213 TI - A new role for astrocytes: beta-amyloid homeostasis and degradation. PMID- 12900214 TI - Phenothiazinium-based photosensitizers: antibacterials of the future? PMID- 12900215 TI - Prospects for regeneration of gastrointestinal epithelia using bone-marrow cells. PMID- 12900216 TI - Getting muscles moving again after botulinum toxin: novel therapeutic challenges. PMID- 12900217 TI - The genetic and molecular basis of epilepsy. AB - In the past decade, studies of large families in which epilepsy has been inherited in an autosomal dominant fashion have revealed several mutated genes, most of which encode ion channel subunits. Despite these exciting findings, only a few families with similar phenotypes have mutations in these known genes. More frustrating has been the genetic research into idiopathic epilepsies with complex inheritance. Although these forms are more common than those with Mendelian inheritance, their unknown mode of inheritance, phenotypic heterogeneity and the uncertainty of the genetic overlap among syndrome subtypes have hampered gene mapping. New techniques of molecular analysis could help the dissection of genes for epilepsies with complex inheritance. Hopefully, in the near future, successful genetic studies will make possible the discovery of new and more targeted anti-epileptic drugs. PMID- 12900218 TI - Targeting cancer cells by exploiting their resistance. AB - Cancer cells are intrinsically resistant to growth arrest and can further acquire multidrug resistance. Current approaches to this problem are intended to reverse, overcome or prevent the drug resistance. However, the resistance of cancer cells can be exploited to kill resistant cells selectively, while sparing sensitive normal cells. As the simplest example, multidrug-resistant cells pump out protectors, such as pharmacological inhibitors of apoptosis. A sequence of at least two agents must include an exclusive protector, which is ineffective in resistant cancer cells, and an inclusive cytotoxic drug, which kills unprotected cells. By abolishing several dose-limiting side effects of chemotherapy, this strategy provides a means to treat selectively most deranged, aggressive and resistant cancers. PMID- 12900219 TI - The molecular basis of myocardial hypertrophy and heart failure. AB - Heart failure (HF) is the inability of the heart to cope with the metabolic demands of the periphery. It is the common end-stage of many frequent cardiac diseases and is characterized by relentless progression. Mechanisms of progression include renal sodium and water retention, neurohumoral activation and alterations of the protein composition (gene programme) of the heart itself. In this review, we explain the often confusing terminology in the subject, briefly touch upon the peripheral mechanisms of HF, and then focus on the changes in the gene programme of the failing heart and the molecular mechanisms leading to them. Understanding the basic processes underlying HF will help uninitiated readers to gain insight into recent novel approaches to its treatment. PMID- 12900221 TI - Methods of quantifying posterior capsule opacification. PMID- 12900223 TI - Needle capsulorhexis: the importance of using an anterior chamber maintainer. PMID- 12900225 TI - Managing after-cataract in pediatric aphakia. PMID- 12900226 TI - Acrysof pupillary capture without pupillary block. PMID- 12900227 TI - Biometry surprises. PMID- 12900228 TI - Multiport-illumination-system-assisted phacoemulsification in eyes with dense vitreous hemorrhage during combined cataract and vitreous surgery. AB - We describe the use of a multiport illumination system to enhance the red reflex during combined phacoemulsification and pars plana vitrectomy in 10 patients with cataract and vitreous hemorrhage. This method improves illumination and visualization of the lens structures and the instruments, which may reduce the risk of complication. To our knowledge, this approach has not been reported. PMID- 12900229 TI - Removal of lens material dropped into the vitreous cavity during cataract surgery using an optical fiber-free intravitreal surgery system. AB - We describe a technique for removing lens fragments dropped into the vitreous cavity during cataract surgery using an optical fiber-free intravitreal surgery system (OFFISS) that was developed for bimanual vitrectomy. The dropped nucleus was removed by 2-port corneal vitrectomy using the OFFISS in 3 patients during complicated cataract surgery. No complications occurred, and good visual recovery was achieved. The OFFISS provided a good view of the fundus without fiber optics and allowed the use of 2 ports for vitrectomy through the corneal wound. This technique is suitable for cataract surgeons who have not been trained in 3-port pars plana vitrectomy. PMID- 12900230 TI - Laser-assisted subepithelial keratectomy enhancement of residual myopia after primary myopic LASEK: six-month results in 10 eyes. AB - PURPOSE: To determine whether residual myopia after primary laser-assisted subepithelial keratectomy (LASEK) can be corrected by repeated LASEK and whether this procedure is safe and effective. SETTING: University Eye Clinic, University of Regensburg, Regensburg, Germany. METHODS: Laser-assisted subepithelial keratectomy was performed in 10 eyes that had had primary LASEK for myopia. The initial mean spherical equivalent (SE) was -6.2 diopters (D) (range -2.5 to -8.0 D), and the residual mean SE was -1.6 D (range -1.0 to -3.0 D). The retreatment ratio was 7.6%. The exposure time of the corneal epithelium to alcohol (20% ethanol) was 30 seconds plus an additional 10 to 15 seconds in cases of strong epithelial adherence. Laser ablation was performed with a Keracor 117 excimer laser (Chiron Technolas). The follow-up after the second LASEK treatment was 6 months. RESULTS: The epithelial flap for repeated LASEK was successfully created in all 10 eyes. The postoperative course in the eyes was uneventful. Six months after LASEK enhancement, the SE refraction was within +/-0.5 D in all eyes. No eye lost more than 1 line of best corrected visual acuity, and the uncorrected visual acuity was 20/20 or better in 8 eyes. Significant haze was not observed after repeated LASEK (mean 0.35 before and 6 months after LASEK enhancement). CONCLUSIONS: Laser-assisted subepithelial keratectomy enhancement appears to be safe and effective for the correction of residual myopia after primary LASEK. Repeated epithelial flap preparation is possible without technical modifications. PMID- 12900231 TI - Microkeratome-assisted lamellar keratoplasty for keratoconus: stromal sandwich. AB - PURPOSE: To evaluate microkeratome-assisted lamellar keratoplasty for the treatment of keratoconus when it is not possible to correct the astigmatic ametropia with contact lenses. SETTING: Ophthalmology Department, School of Medicine, Gazi University, Ankara, Turkey. METHODS: This prospective study comprised 9 eyes of 7 keratoconus patients with contact lens intolerance. The donor cornea was prepared with a microkeratome and punched with a 7.25 mm or 7.50 mm trephine. Following the creation of a standard 9.0 mm corneal flap in the host cornea, the donor stromal button was implanted under this corneal flap like a sandwich. Transepithelial photorefractive keratectomy or laser in situ keratomileusis was performed when the corneal topography and refraction stabilized by the end of the sixth postoperative month. Follow-up ranged from 7 to 22 months. RESULTS: All patients gained 5 or more lines (mean 7.2 lines +/- 1.6 [SD]), and no patient lost a line of vision. The mean corneal thickness was 432.7 +/- 36.1 micrometers preoperatively and 578.1 +/- 45.1 micrometers after refractive surgery. CONCLUSION: The early visual results of this surgical technique are promising and seem to be comparable to those with penetrating keratoplasty. PMID- 12900232 TI - Postoperative endophthalmitis: establishment and results of a national registry. AB - PURPOSE: To determine the incidence, clinical presentation, etiology, and outcomes of postoperative endophthalmitis in Norway and to explore the potential of and establish a platform for improving diagnostics, prophylaxis, and treatment of postoperative endophthalmitis. SETTING: All ophthalmic surgical units and relevant microbiology laboratories in Norway. METHODS: A national registry of cases of postoperative endophthalmitis was established in 1996. All ophthalmic surgery units in Norway were asked to return forms including preoperative, perioperative, and postoperative data for each case of endophthalmitis occurring between January 1996 and December 1998 after any intraocular surgery irrespective of the time between the surgery and the onset of endophthalmitis symptoms. All microbiology laboratories in Norway were asked to return questionnaires regarding their routine handling procedures of eye specimens in cases of endophthalmitis. RESULTS: From 1996 to 1998, 111 suspected endophthalmitis cases were reported, all after cataract surgery. Eighty cases were culture positive, 75 with gram positive bacteria, 4 with gram-negative bacteria, and 1 with Candida albicans. Depending on the definition, the incidence of postoperative endophthalmitis was between 0.11% and 0.16%. Thirteen percent of cases had "delayed" endophthalmitis. Twenty-nine percent of eyes had an outcome of permanent amaurosis or light perception visual acuity; 56% (47/84) retained or improved their categorized visual acuity in the affected eye compared to preoperatively. Positive bacterial growth was associated with a worse visual outcome (P =.008). The degree of inflammation when endophthalmitis was diagnosed and the visual outcomes were worse in cases with growth of streptococci than in cases with growth of staphylococci (P =.009 and P<.001, respectively). The questionnaire to the microbiology laboratories revealed a lack of consensus on how to handle the specimens. CONCLUSIONS: Postoperative endophthalmitis remains a serious complication of intraocular surgery, although the prognosis depends greatly on the microbe isolated. Common guidelines should be established regarding clinical and microbiological diagnosis and treatment. Further improvement of the registry would make it a suitable platform for evaluating prophylactic treatments. PMID- 12900233 TI - Simultaneous bilateral cataract surgery. AB - PURPOSE: To evaluate the safety and outcomes of simultaneous bilateral cataract surgery (SBCS). SETTING: York Finch Eye Associates and Humber River Regional Hospital, Toronto, Ontario, Canada. METHODS: This retrospective study reviewed the results of 1020 consecutive patients (2040 eyes) who had SBCS by endolenticular phacoemulsification through a clear corneal incision on the corneal steep axis with foldable posterior chamber intraocular lens implantation. The surgeries were performed by the same surgeon from January 1996 to January 2002 as 2 consecutive independent procedures under topical and intracameral anesthesia. Outcome measures included intraoperative and postoperative complications, postoperative uncorrected and best spectacle-corrected visual acuities, refractive error, and patient satisfaction. RESULTS: Complications were few and would not likely have been prevented had the surgery been performed monocularly. CONCLUSIONS: Simultaneous bilateral cataract surgery did not lead to an increased incidence of intraoperative or postoperative complications. The visual acuity results were good, and the patients were pleased. No complications were observed that could be attributed to the procedures being done bilaterally. PMID- 12900234 TI - Slitlamp stretching of the corneal flap after laser in situ keratomileusis to reduce corneal striae. AB - PURPOSE: To examine the efficacy of treating immediate postoperative corneal striae and poor flap alignment by stretching the corneal flap at the slitlamp with a cotton-tip applicator and compare the results using this technique with those using the conventional technique of refloating the flap with balanced salt solution (BSS(R)). SETTING: TLC Laser Eye Center, Garden City, New York, USA. METHODS: Charts of 7154 eyes having laser in situ keratomileusis (LASIK) were reviewed retrospectively. During the 11 months prior to January 1, 2000, 3516 eyes had LASIK. Patients with flap striae or a malpositioned flap at the immediate slitlamp evaluation were treated by refloating the flap (Group A). During the 11 months subsequent to January 1, 2000, 3638 eyes had LASIK. Patients with flap striae or a malpositioned flap at the immediate slitlamp evaluation were treated by flap stretching with a cotton-tip applicator (Group B). The number of eyes that required retreatment for flap striae was analyzed. RESULTS: Twenty-nine eyes in Group A (0.82%) required retreatment for visually significant flap striae after day 1, and 11 eyes in Group B (0.30%) required retreatment. A loss of best corrected visual acuity occurred in 3 Group A eyes and 2 Group B eyes. CONCLUSIONS: During the immediate postoperative period, stretching the flap with a cotton-tip applicator was a simple, safe, and effective technique for reducing visually significant flap striae. Unlike refloating the flap with BSS, flap stretching at the slitlamp does not require additional anesthesia, exposure time, or dehydration of the ocular surface. PMID- 12900235 TI - Changes in corneal curvature in accommodation. AB - PURPOSE: To determine the changes in corneal curvature in accommodation. SETTING: Department of Ophthalmology, St. Luke's International Hospital, Tokyo, Japan. METHODS: Twenty-eight eyes of 14 healthy volunteers with a mean age of 38.9 years (range 28 to 65 years) were enrolled in this prospective study. Corneal topographies were taken before and during accommodation. Subjects achieved maximal accommodation by gazing at a target letter "A" in front of the eye. Keratometric values (K-values) obtained by topographies before and during accommodation were compared. RESULTS: Steepened corneal curvatures during accommodation were observed with the difference-plot view. Maximum K-values for the central 3.0 mm, 5.0 mm, and 7.0 mm corneal diameters significantly increased by a mean of 0.62 diopters (D) +/- 0.83 (SD) (P =.0005, paired t test), 0.60 +/- 0.90 D (P =.0015), and 0.72 +/- 0.65 D (P<.0001), respectively. Respective minimum K-values significantly increased by a mean of 0.62 +/- 0.83 D (P=.0005), 0.67 +/- 0.68 D (P<.0001), and 0.64 +/- 0.62 D (P<.0001). No statistically significant relationship was found between age and change in K-values. CONCLUSION: The results suggest that changes in corneal curvature in accommodation participate in the mechanism of accommodation. PMID- 12900236 TI - Superficial keratectomy with mitomycin-C for the treatment of Salzmann's nodules. AB - PURPOSE: To evaluate the success of treatment and prevention of recurrence of Salzmann's nodules by superficial keratectomy with intraoperative mitomycin-C (MMC). SETTING: Price Vision Group, Indianapolis, Indiana, USA. METHODS: This retrospective consecutive case series comprised 30 eyes of 25 patients who had superficial keratectomy with MMC between January 1997 and June 2000 at a large tertiary-care center. RESULTS: Twenty-four patients reported improvement in their symptoms. The remaining patient lost 1 line of visual acuity. Ninety-seven percent of eyes maintained or improved best corrected visual acuity. No patient had a recurrence of Salzmann's nodular degeneration over a mean follow-up of 28 months +/- 15 (SD) (range 4.0 days to 4.1 years). CONCLUSION: Superficial keratectomy with MMC appears to be a valid and safe method for treating and preventing the recurrence of Salzmann's nodular degeneration. PMID- 12900237 TI - Clinical study of the 1CU accommodating intraocular lens. AB - PURPOSE: To compare the near functional capacities of patients with an accommodating intraocular lens (IOL) with those of patients with a conventional monofocal IOL. SETTING: Department of Ophthalmology, University of Chieti, Chieti, Italy. METHODS: This prospective double-blind case-control study comprised 42 eyes that had phacoemulsification and implantation of 1 of 2 types of IOLs: HumanOptics accommodating 1CU(R) (study group) and Eurocrystal IFP 3G 6.00 (control group). The main outcome measures were subjective refraction, uncorrected distance acuity, best corrected distance acuity, distance corrected near acuity at 40 cm, best corrected near acuity at 40 cm, and subjective amplitude of accommodation. Patients were examined 7, 30, 90, and 180 days after surgery. RESULTS: Postoperatively, both groups had excellent uncorrected distance acuity, best corrected distance acuity, and best corrected near acuity. In the study group, the mean distance corrected near acuity (Jaeger) was 5.43 +/- 0.98 (SD) (range 4 to 7) at 7 days, 2.33 +/- 0.48 (range 2 to 3) at 1 and 3 months, and 3.66 +/- 2.12 (range 2 to 7) at 6 months. In the control group, the mean distance corrected near acuity was 7.43 +/- 0.50 (range 7 to 8) during the entire follow-up. The differences between the groups was statistically significant (P<.001). The mean amplitude of accommodation was 0.00 diopter (D) in the control group and 1.14 +/- 0.44 D (range 0.75 to 2.00 D) in the study group at 7 days, 2.36 +/- 0.28 D (range 2.00 to 2.75 D) at 30 and 90 days, and 1.90 +/- 0.77 D (range 0.75 to 2.75 D) at 6 months. CONCLUSIONS: The 1CU accommodating IOL provided better useful spectacle-free near visual acuity than the conventional monofocal IOL. However, the accommodating mechanism can play a role in capsule fibrosis. PMID- 12900238 TI - Corneal thickness measurements: scanning-slit corneal topography and noncontact specular microscopy versus ultrasonic pachymetry. AB - PURPOSE: To compare central corneal thickness measurements taken with 3 pachymetry systems: Orbscan scanning-slit corneal topography/pachymetry, Topcon SP2000P noncontact specular microscopy, and Tomey ultrasonic pachymetry. SETTING: Multicenter study, Tokyo, Japan. METHODS: In 216 healthy eyes of 114 subjects, scanning-slit topography, noncontact specular microscopy, and ultrasonic pachymetry were used in that sequence to record central corneal thickness. In another 20 healthy eyes of 13 subjects, 2 sets of measurements were repeated for each pachymetry to assess repeatability. RESULTS: The mean central corneal thickness was compatible between scanning-slit topography (546.9 micrometers +/- 35.4 [SD] ) and ultrasonic pachymetry (548.1 +/- 33.0 micrometers); however, noncontact specular microscopy gave a significantly smaller mean (525.3 +/- 31.4 micrometers) than the other 2 tests (P<.001, Tukey multiple comparison). There were significant linear correlations between scanning-slit topography and noncontact specular microscopy (r = 0.846, P<.001), noncontact specular microscopy and ultrasonic pachymetry (r = 0.897, P<.001), and ultrasonic pachymetry and scanning-slit topography (r = 0.852, P<.001). Noncontact specular microscopy tended to show the best repeatability; however, the difference was not statistically significant (P =.663, repeated-measure analysis of variance). CONCLUSIONS: Corneal thickness readings were comparable between scanning-slit topography and pachymetry; noncontact specular microscopy gave significantly smaller values. The measurements of the 3 methods showed significant linear correlations with one another. All methods provided acceptable repeatability of measurements. PMID- 12900239 TI - Effect of contact lens wear on central corneal thickness measurements. AB - PURPOSE: To compare central corneal thickness measurements as a function of contact lens wear in patients presenting for refractive surgery. SETTING: Gimbel Eye Centre, Saskatoon, Saskatchewan, Canada. METHODS: A retrospective study of charts yielded 634 eyes meeting the inclusion criterion. Pachymetry measurements of spectacle-wearing control eyes were compared to those of full-time soft contact lens wearers and rigid gas-permeable contact lens wearers. All measurements were taken with the DGH ultrasonic pachymeter by a single observer. RESULTS: Soft contact lens wearers had a mean reduction in corneal thickness measurements of 22 micrometers compared to the control population (P =.00001). Rigid gas-permeable lens wearers had a reduction of 24 micrometers compared to the control population (P =.00001). CONCLUSION: There was a clinically significant reduction in corneal thickness measurements associated with full-time contact lens wear. PMID- 12900240 TI - Actual and theoretical risks for visual loss following use of the implantable contact lens for moderate to high myopia. AB - PURPOSE: To estimate the actual and theoretical risks for visual loss associated with use of the implantable contact lens (ICL). SETTING: Multicenter clinical trial data. METHODS: An established model for estimating the risk for visual loss associated with retinal detachment (RD) following clear lens extraction (CLE) was modified to be applicable to the ICL. The risk for developing RD and the outcomes of RD surgery based on current lens extraction and RD repair techniques were updated. Non-RD-related risks for pseudophakia based on United States Food and Drug Administration (FDA) guidelines, actual visual loss data from the FDA ICL study, and calculated risks assuming a large future risk (10 times the current risk) of cataract development were also added. RESULTS: The additional risk for visual loss due to cataract surgery is estimated at 0.19% and for subsequent RD assuming 10 times the current rate of cataract is 0.37%. Using an ultraconservative (high) model and adding the actual loss observed with the ICL, the total risk is 1.39%. The risk increases by 0.5% to 0.6% with each 10% increase in cataract incidence (eg, 10% to 20%, 20% to 30%). The risk for visual loss with CLE and/or laser in situ keratomileusis/photorefractive keratectomy in the same range of myopia is possibly 3.0% to 6.5%. CONCLUSIONS: The ICL represents a viable alternative to currently available refractive procedures based on an analysis of the risk for visual loss. PMID- 12900241 TI - Retinal detachment after phacoemulsification cataract extraction. AB - PURPOSE: To estimate the incidence of retinal detachment (RD) after phacoemulsification cataract extraction in an unselected group of patients. SETTING: Herlev University Hospital, Copenhagen, Denmark. METHODS: This retrospective review comprised 6521 cataract patients who had phacoemulsification over a 3-year period. The incidence of postoperative RD and the risk factors were assessed. RESULTS: During the 15- to 52-month observation period, 22 cases of RD in 21 patients were observed, with an estimated risk (Kaplan-Meier) of 0.41% (95% confidence limit, 0.25%-0.57%) at 52 months. The results confirmed recognized preoperative risk factors for RD such as an axial length greater than 25.0 mm (P<.001), age younger than 65 years (P<.001), and male sex (P<.02). In 18 eyes, the retina was reattached after 1 operation. CONCLUSIONS: The incidence of RD after phacoemulsification cataract extraction in an unselected group of patients was low. A long axial length, young age, and male sex were confirmed as risk factors. PMID- 12900242 TI - Laser cataract surgery: technique and clinical results. AB - PURPOSE: To study the effectiveness of an erbium:YAG (Er:YAG) laser in removing nuclei of various densities. SETTING: University clinic. METHODS: This nonrandomized prospective clinical study included 147 consecutive cataractous eyes: Group 1 (n = 71) had conventional ultrasound, and Group 2 (n = 76) had surgery with the Er:YAG laser. The 2 groups were further classified by cataract density. Patients were examined 6 months postoperatively to evaluate surgical time, intraocular pressure (IOP), pachymetry, endothelial cell loss, and visual acuity. RESULTS: With ultrasound cataract surgery, there were no significant changes in IOP compared with the preoperative measurements. Laser treatment of 3+ and 4+ cataracts resulted in IOP increases of 12.6% and 18.2%, respectively, compared with 1+ and 2+ cataracts (P<.001). Slight endothelial cell losses occurred in all cases. With ultrasound, the mean decrease was 2.8%, and with laser surgery, the results were similar except for a significant decrease of 13.4% with 4+ cataracts. There was a correlation between pachymetry and surgical time when the trauma related to surgical time and hydrodynamic flow in the anterior chamber was assessed. With ultrasound, the visual acuity was similar with all types of cataract; with the laser, similar normal values were seen with 1+ and 2+ cataracts but decreased values were seen with 3+ cataracts and lower values with 4+ cataracts-0.43 compared to 0.84 best corrected visual acuity with same-density cataracts treated with ultrasound (P<.0005). Patients with 4+ cataracts treated with the laser developed more complications than those treated with ultrasound. CONCLUSIONS: The Er:YAG laser was as effective and reliable as ultrasound in removing soft and medium-density cataracts. With dense cataracts, the surgical length produced more trauma and complications during laser treatment than during ultrasound treatment. PMID- 12900243 TI - Intraocular lens power calculation after refractive surgery. AB - PURPOSE: To analyze the results of phacoemulsification cataract surgery in eyes that had had refractive surgery and to compare the predictability of various methods of intraocular lens (IOL) power calculation. SETTING: Instituto de la Vision, Buenos Aires, Argentina. METHODS: The study involved 7 cases that had phacoemulsification after radial keratotomy or laser in situ keratomileusis. The spherical equivalent (SE) and visual acuity were evaluated preoperatively and postoperatively to assess the changes before cataract development. The IOL power calculated with conventional keratometry (CK), adjusted keratometry, the clinical history method (CHM), corneal topography (CT), and the contact lens method (CLM) was compared with the final refractive and keratometric results measured with the BackCalcs (Holladay(R) IOL Consultant Program, Holladay Consulting, Inc.) to assess the accuracy and predictability of each method. RESULTS: The mean SE was 4.82 diopters (D) +/- 5.13 (SD) before phacoemulsification and +0.19 +/- 1.01 D after phacoemulsification, and the mean best corrected visual acuity was 0.39 +/- 0.07 (20/50) and 0.80 +/- 0.06 (20/25), respectively. CONCLUSIONS: Post phacoemulsification refraction in cases with previous refractive surgery appeared to be predictable when the appropriate calculation method was applied. When all the data were available, the CHM provided the best results. Adjusted keratometry and CT seemed to be more accurate than CK and the CLM. PMID- 12900244 TI - Intraocular lens calculations in patients with corneal scarring and irregular astigmatism. AB - PURPOSE: To compare various methods of estimating corneal power for intraocular lens (IOL) calculation in patients with irregular corneal astigmatism. SETTING: Pepose Vision Institute, St. Louis, Missouri, USA. Case reports and review of the medical literature. RESULTS: Two patients with irregular corneal astigmatism had an IOL exchange after a "surprise" post-cataract-surgery refraction. In the first case, the patient had a post-cataract-surgery refraction of +5.50 -0.75 x 69 and in the second case, a refraction of -7.00 -1.00 x 180. The central corneal power before IOL exchange was assessed using manual keratometry, various computerized videokeratography curvature and power maps, and contact lens overrefraction. The total axial power map (Orbscan(R), Bausch & Lomb), total optical power map (Orbscan), and contact lens overrefraction method provided the most accurate estimates of central corneal power in these 2 patients. CONCLUSION: Computerized scanning-slit videokeratography, which analyzes the anterior and posterior surfaces of the cornea, and the contact lens overrefraction method gave good estimations of corneal power in patients with irregular corneal astigmatism. This type of analysis may improve the accuracy of IOL calculation in patients with corneal pathology and irregular astigmatism. PMID- 12900245 TI - Intraocular lens power calculation formulas in Chinese eyes with high axial myopia. AB - PURPOSE: To compare the accuracy of intraocular lens (IOL) power calculation formulas in Chinese eyes with high axial myopia. SETTINGS: Department of Ophthalmology, Tuen Mun Hospital, Hong Kong, China. METHODS: This retrospective study reviewed 125 Chinese patients with high myopia and axial lengths (ALs) longer than 25.0 mm who had cataract surgery during the year 2000. Eyes with pathology or operative complications affecting the refractive status and those with missing data were excluded. In each case, the power of the implanted IOL was used to calculate the predicted postoperative refractive error by 4 IOL power calculation formulas: SRK II, SRK/T, Holladay 1, and Hoffer Q. The predictive accuracy of the formulas was analyzed by comparing the difference between the "actual" and "predicted" postoperative refractive errors. The Student t test was used for statistical analysis. The performance of the formulas in subcategories of long AL was also tested. RESULTS: One hundred twenty-five eyes (110 patients) were studied. Thirty-seven eyes (29.6%) were excluded. The Hoffer Q, Holladay 1, and SRK/T formulas showed a slight tendency toward resultant hyperopia, with a mean of +0.36 diopters (D), +0.53 D, and +0.74 D, respectively. The SRK II caused the largest hyperopic error, with a mean of +1.47 D. All 4 formulas caused a refractive error shift toward myopia in the subcategories of AL >28.0 mm, minus power IOL, and extracapsular cataract extraction (ECCE). CONCLUSIONS: In Chinese eyes with high axial myopia with an AL longer than 25.0 mm, the 4 formulas caused a slight postoperative hyperopic refractive error that was less in eyes with a minus-power IOL or an AL longer than 28.0 mm and in those that had ECCE. The Hoffer Q formula provided the best predictive result, and Holladay 1 and SRK/T were comparable in IOL power calculation. The SRK II was the least accurate in all subgroups. PMID- 12900246 TI - Effect of visual acuity on biometry prediction error after cataract surgery. AB - PURPOSE: To determine the effect of visual acuity on biometry prediction error. SETTING: Postgraduate teaching hospital. METHODS: The study was an observational case series of the first operated eye of 2149 consecutive patients who had cataract surgery. Biometry prediction error was defined as the difference between the planned refraction determined by biometry and the spherical equivalent of the final refraction. The principal outcome measure was the percentage of eyes within +/-1.00 diopter (D) of the intended refraction. This outcome was calculated for patients with visual loss caused by cataract alone, defined as eyes with a postoperative acuity of 6/6 or better, and eyes with different levels of postoperative visual acuity. RESULTS: Complete data were available for 1978 eyes (92%). Of these, 1438 (73%) were within +/-1.00 D of the intended refraction. When the final corrected visual acuity was 6/6 or better, the percentage of eyes within +/-1.00 D of the predicted refraction was at least 78%. When the preoperative acuity was worse than 6/60, 74% in the counting fingers group and 69% in the hand movements group with a final corrected acuity of 6/6 or better were within +/-1.00 D. This trend bordered on statistical significance (P =.05). There was a rapid reduction in the percentage of eyes within +/-1.00 D of the intended refraction as the postoperative acuity decreased because of ocular comorbidity, with a statistically significant difference between eyes with a postoperative acuity of 6/6 or better and each group with an acuity of 6/9 or worse (P<.01). CONCLUSIONS: In eyes without visually significant ocular comorbidity, cataract must reduce vision to 6/60 or worse before there is an increase in biometry prediction error. In contrast, ocular comorbidity that reduces the postoperative acuity causes an early and marked increase in biometry prediction error. PMID- 12900247 TI - Endophthalmitis secondary to corneal sutures: series of delayed-onset keratitis requiring intravitreal antibiotics. AB - PURPOSE: To describe 5 cases of endophthalmitis secondary to corneal sutures. SETTING: Department of Ophthalmology, University College Hospital, Galway, Ireland. METHODS: A retrospective review was done of 5 patients who presented over a period of 19 months with suture-related bacterial keratitis that progressed to endophthalmitis. Intravitreal antibiotics were used to control the infection. RESULTS: Despite intensive topical broad-spectrum antibiotic treatment for suture-related infective keratitis, the infection progressed to endophthalmitis in all 5 patients. Intravitreal antibiotics resolved the infection. CONCLUSIONS: Corneal sutures are a risk factor for infective keratitis irrespective of their site. Suture-related keratitis may progress rapidly to endophthalmitis despite intensive topical antibiotics. Intravitreal antibiotics are indicated when endophthalmitis is present. PMID- 12900248 TI - Confocal microscopic evaluation of particles at the corneal flap interface after myopic laser in situ keratomileusis. AB - PURPOSE: To investigate the appearance and origin of interface particles in the cornea after myopic laser in situ keratomileusis (LASIK) using the confocal microscope. SETTING: Department of Optometry and Neuroscience, UMIST, Manchester, United Kingdom. METHODS: An in vivo slit-scanning, real-time confocal microscope (Tomey ConfoScan P4) fitted with an Achroplan(R) 40X/0.75 NA immersion objective (Zeiss) was used to examine the morphology of the central cornea in 6 patients (12 eyes) at the initial visit (before surgery) and 1 week and 1, 3, and 6 months after LASIK for myopia. RESULTS: In all eyes, several interspersed particles of variable size and brightness were observed 1 week after surgery at the flap interface. The overall density of these particles decreased with time (F = 14.34, P =.01). For analysis, the particles were divided by density into high brightness and low brightness. The low-brightness particles significantly decreased with time (F = 13.26, P =.02). The high-brightness particles remained constant in density at all postoperative visits (F = 1.3, P =.15). CONCLUSIOINS: The particles of low reflectivity may represent 1 or more types of remains. It is unclear whether the low-reflectivity particles will eventually disappear. The high-reflectivity particles may represent fine metallic debris arising from the mechanical drive mechanism and/or the disposable knife of the microkeratome. If they are metallic and magnetic, they could be cleared during surgery (immediately before flap replacement) using a specially adapted magnet. PMID- 12900249 TI - Conversion from trabeculectomy to deep sclerectomy. Prospective study of the first 44 cases. AB - PURPOSE: To evaluate the outcomes of conversion from trabeculectomy to deep sclerectomy with placement of a drainage implant. SETTING: Department of Ophthalmology, Hospital of Buskerud, Drammen, Norway. METHODS: Deep sclerectomy was performed in 41 patients (44 eyes) from November 1999 to January 2002. An absorbable implant (AquaFlow((R)), Staar Surgical) and a nonabsorbable implant (T.Flux((R)), IOLTech Laboratories) were used in 75% and 25% of eyes, respectively. Patients with open-angle glaucoma who needed filtering surgery were enrolled consecutively in a prospective study. Patients with 2 or more previous filtering surgeries or neovascular glaucoma were excluded, as were children. Of the original 48 eyes, accidental perforation of the trabeculo-Descemet's membrane occurred in 8.3%; these eyes were excluded from the study. The severity of optic nerve damage was considered in the decision to start therapy. An intraocular pressure (IOP) above 22 mm Hg was always treated. RESULTS: After a mean follow-up of 12.9 months +/- 7.1 (SD), 61.4% of eyes were controlled without therapy. When eyes with previous surgery were excluded, the rate increased to 66.7% after a mean follow-up of 13.8 +/- 7.2 months. The mean IOP preoperatively and after 12 months was 27.9 +/- 6.9 mm Hg and 14.6 +/- 3.5 mm Hg, respectively (P<.001). The mean number of medications decreased from 3.4 +/- 1.0 preoperatively to 0.6 +/- 1.1 12 months postoperatively (P<.001). A reoperation was required in 2 cases (4.5%). There were no complications related to hypotony or other significant complications. CONCLUSIONS: This study shows that conversion from trabeculectomy to deep sclerectomy with placement of an implant is safe. The postoperative IOP reduction was acceptable, and no significant complications occurred. PMID- 12900250 TI - Surface scattering in implanted hydrophobic intraocular lenses. AB - PURPOSE: To describe surface scattering, a new phenomenon in polyacrylic intraocular lenses (IOLs), and discuss the possible cause of this finding. SETTING: Showa University, Fujigaoka Hospital, Yokohama, Kanagawa, Japan. METHODS: This observational case series involved 40 patients who received the AcrySof(R) IOL (Alcon Laboratories, Inc.) as part of a clinical trial in 1991. Four patients (5 eyes) were evaluated. Thirty-three patients had the same surgery in 1999, and the findings in these patients were compared with those in the earlier patients. Lens surface scattering was evaluated by area densitometry using Scheimpflug photography. No statistical analysis was done because of the small sample size. RESULTS: The 5 IOLs implanted in 1991 showed various degrees of surface scattering. The degree of scattering was greater than that in the 1999 group. There was no degradation in visual performance and no manifestation of intraocular inflammation or other adverse effect from the IOLs in either group. Attempts to remove foreign material on the IOL using the neodymium:YAG (Nd:YAG) laser were unsuccessful. CONCLUSIONS: The surface scattering did not appear to be due to deposition of foreign material on the IOL surface. Densitometry and the results of Nd:YAG laser treatment suggest that the changes occurred in the near surface layer. Phase separation of water near the IOL surface in the inherently hydrophobic material may explain the phenomenon. Newer AcrySof IOLs appear to have less surface scattering. PMID- 12900251 TI - Electron microscopy of the epithelial flap created by ethanol treatment in the rabbit corneal epithelium. AB - PURPOSE: To assess the effect of corneal debridement with 25% ethanol on rabbit corneal epithelium by electron microscopy. SETTING: Department of Ophthalmology, Osaka University School of Medicine, Osaka, Japan. METHODS: Rabbit corneas were deepithelialized by applying 25% ethanol for 3 minutes, and a hinged epithelial flap was created and repositioned. Ten corneas were evaluated immediately after the epithelial debridement procedure and at 1, 3, and 7 days. Histological changes were evaluated using scanning and transmission electron microscopy. RESULTS: Corneas treated with ethanol showed a decrease in microvilli, breaks in intercellular junctions, epithelial cell edema, and damage in basement membrane hemidesmosomes. These toxic changes recovered to an almost normal appearance after 24 hours and rapidly improved over the following week. An increase in desquamated cells was observed after debridement; this returned to normal after 1 week according to the normal cellular cycle. Wing cells showed no damage at any stage. CONCLUSIONS: Although ethanol appeared to have a toxic effect on rabbit epithelial cells, the effect did not persist over time. The first signs of recovery were observed 24 hours after debridement, and full recovery was observed over the following week. The ethanol path to the central basement membrane appeared to be centripetally from a round cut through the basement membrane, leaving the wing cells intact. PMID- 12900252 TI - Corneal power after refractive surgery for myopia: contact lens method. AB - PURPOSE: To clarify the theoretical background of the rigid contact lens overrefraction (CLO) method to determine corneal power after corneal refractive surgery. SETTING: University Eye Clinic, University of Wurzburg, Wurzburg, Germany. METHODS: Using paraxial geometrical optics, the measurement situation for the contact lens method was analyzed and the definitions of corneal refractive power were reviewed. Based on the theoretical Gullstrand eye, model eyes were constructed, representing 1 emmetropic and 2 myopic eyes (primary refraction -5.21 diopters [D] and -10.25 D, respectively) before and after photorefractive keratectomy and laser in situ keratomileusis. In these eyes, the application of the CLO was mathematically simulated using Gaussian thick-lens optics and commercial ray-tracing software. RESULTS: The CLO method measured neither the equivalent (total) power nor the vertex (back) power of the cornea but rather the quantity 336/R(1C) (R(1C) = anterior corneal radius). Based on these results and the Gullstrand eye, new formulas are proposed to derive the equivalent power and vertex power of the cornea by the CLO method. CONCLUSIONS: Depending on whether intraocular lens calculation formulas are based on equivalent (total) corneal power or vertex corneal power, the respective new formulas for the CLO method should be applied in patients after corneal refractive surgery. An increase in prediction accuracy of the refractive outcome is expected. PMID- 12900253 TI - Practice styles and preferences of ASCRS members--2002 survey. AB - A survey of the practice styles and preferences of members of the American Society of Cataract and Refractive Surgery (ASCRS) with a United States ZIP code was performed in August 2002. Approximately 18% (1056) of the 5816 questionnaires mailed were returned. Three profile questions were used to cross-tabulate: age of the respondent, geographic location, and volume of cataract surgery per month. The refractive surgical questions were cross-tabulated for the volume of laser in situ keratomileusis (LASIK). Data in this survey were compared with those in previous surveys of ASCRS members. PMID- 12900254 TI - Intracorneal rings for the correction of pellucid marginal degeneration. AB - We report a case of Intacs(R) (KeraVision) implantation for the correction of pellucid marginal degeneration (PMD). Preoperatively, the patient's uncorrected visual acuity (UCVA) was 0.05, the best spectacle-corrected visual acuity (BSCVA) was 0.1, and the refraction was -2.00 -7.00 x 90 in the right eye. The flattest meridian (K1) measured 43.8@104 and the steepest meridian (K2), 51.3@14. Ultrasound pachymetry revealed a thin cornea of 420 micrometers. One month postoperatively, the UCVA was 0.2, the BSCVA was 0.5, and the refraction was 8.00 -7.00 x 50. Contact lens best corrected visual acuity was 0.9; 6 months postoperatively, it improved to 1.0. The use of Intacs to treat PMD seems to be viable and improves visual acuity. The residual error can be corrected with contact lenses. PMID- 12900255 TI - Fibrin glue for the prevention of epithelial ingrowth after laser in situ keratomileusis. AB - We present 3 cases of recurrent epithelial ingrowth following laser in situ keratomileusis. All patients consented to flap lifting, epithelial ingrowth removal, and placement of fibrin glue (Tisseel(R) VH, Baxter) at the flap edges. Two of 3 eyes had no evidence of recurrent epithelial ingrowth within a relatively short follow-up. One eye that also had had radial keratotomy had a small nest of stable epithelial cells that had not progressed. Fibrin glue may be a useful adjunct to prevent recurrent epithelial ingrowth. There were no complications associated with its use. PMID- 12900256 TI - Phototherapeutic keratectomy with amniotic membrane for severe subepithelial fibrosis following excimer laser refractive surgery. AB - We report 2 patients who developed subepithelial opacities and myopic regression after photorefractive keratectomy (PRK) and laser-assisted subepithelial keratectomy. Both cases were treated successfully with manual debridement of the epithelium, phototherapeutic keratectomy, and PRK with amniotic membrane application. PMID- 12900257 TI - Corneal edema resolution after "descemetorhexis". AB - A 79-year-old woman had left eye phacoemulsification complicated by inadvertent excision of Descemet's membrane (DM). One day postoperatively, the visual acuity was counting fingers with diffuse corneal edema. The patient was treated medically, with gradual resolution of the corneal edema over a 6-month period. The torn edge of DM could be visualized as the edema cleared, and no endothelial cell count could be obtained centrally. Approximately 1 year postoperatively, the central endothelial cell count was 2114 cells/mm(2) in the right eye and 827 cells/mm(2) in the left eye. The decreased cell count, increased polymegethism, and pleomorphism suggested that endothelial cells migrated to cover the stromal surface area in the absence of DM. PMID- 12900258 TI - Hemorrhagic Descemet's membrane detachment after viscocanalostomy. AB - We report a case of hemorrhagic detachment of Descemet's membrane after viscocanalostomy. A 60-year-old man with pseudoexfoliative glaucoma had an uneventful viscocanalostomy in the right eye. On the first postoperative day, a 5.0 mm x 5.0 mm Descemet's membrane detachment was present in the superior nasal quadrant. The space between Descemet's membrane and the stroma was filled with a hemorrhage. The hemorrhage resorbed by 6 months postoperatively, and Descemet's membrane completely reattached without surgical manipulation. The final visual acuity dropped from 20/40 to 20/100, final intraocular pressure was 20 mm Hg with 1 medication, and a paracentral fibrinoid scar persisted 18 months after surgery. Hemorrhagic detachment of Descemet's membrane can cause visual acuity deterioration and should be recognized as a rare complication of viscocanalostomy. PMID- 12900259 TI - Filtering blebs at the site of sutured posterior chamber intraocular lenses. AB - A 78-year-old man with a traumatic giant retinal tear and phacodonesis had 3-port pars plana vitrectomy (3PPPV), lensectomy, and sutured posterior chamber intraocular lens (IOL) implantation. Two years after surgery, a filtration bleb was noted at 1 of the suture sites. In another case, a 32-year-old man with lens subluxation secondary to Marfan's syndrome had 3PPPV, lensectomy, and sutured posterior chamber IOL implantation. Two months after surgery, a filtration bleb was noted at 1 of the suture sites. Sutured posterior chamber IOL implantation is 1 of the few instances in which there is virtually a full-thickness suture through the sclera. We presume the filtering bleb formed as a direct result of the permanent passage created from the posterior chamber to the subconjunctiva due to presence of the suture. Presence of a filtering bleb can lead to complications including endophthalmitis. PMID- 12900260 TI - Management of bipseudophakia with implantation of an iris claw lens. AB - We discuss a case of bipseudophakia with secondary dislocation of a posterior chamber intraocular lens (IOL) into the anterior chamber that was managed by explantation of the anterior and posterior chamber IOLs and implantation of an iris claw IOL. PMID- 12900261 TI - Isolated microspherophakia with optic disc colobomata. AB - We describe a case of isolated bilateral microspherophakia with optic disc colobomata. A satisfactory outcome was achieved following uneventful lensectomy and scleral fixation of an intraocular lens. PMID- 12900262 TI - Visual experiences of cataract surgery. PMID- 12900263 TI - AB interno side ports in complicated phacoemulsification and pediatric cataract surgery. PMID- 12900265 TI - Whole-body analysis of T cell responses. AB - In vivo methods to detect antigen-specific T cell responses at the single-cell level have greatly increased our knowledge of how the immune system works. However, most of these approaches have been confined to analysis of lymphoid tissues. Recently, the development of imaging techniques capable of simultaneously monitoring all the tissues of the body has led to the realization that antigen-experienced T cells reside in non-lymphoid tissues and may play a vital role in protecting the host against pathogens. Therefore, single-cell imaging at the level of the whole organism is needed to fully understand the dynamics of protective immunity. PMID- 12900266 TI - Real-time imaging of lymphocytes in vivo. AB - New preparations, fluorescent probes and imaging techniques are providing the means to observe the behavior of cells in the tissue environment of lymphoid organs. In particular, when combined with two-photon laser microscopy, intravital imaging of surgically exposed lymph nodes provides a unique view of lymphocyte migration and antigen presentation as it occurs within the living animal. The view is emerging that lymphocytes migrate randomly within lymphoid organs, and that lymphocyte contact with antigen-presenting cells may be a stochastic process rather than one guided by chemokine gradients. PMID- 12900267 TI - Micro-PET imaging and small animal models of disease. AB - Positron emission tomography (PET) has been used clinically to measure enzyme reactions, ligand-receptor interactions, cellular metabolism and cell proliferation. Until recently, however, PET has not been suitable for small animal models because of resolution limitations. Development of micro-PET instrumentation for small animal imaging and the availability of positron emitting tracers has made this technology accessible for the non-invasive, quantitative and repetitive imaging of biological function in living animals. The development of new probes and positron-imaging based reporter genes has extended micro-PET applications to investigations of metabolism, enzyme activity, receptor ligand interactions, protein-protein interactions, gene expression, adoptive cell therapy and somatic gene therapy. Because small animal PET is immediately extrapolatable to the clinic, laboratory advances should rapidly be translated to clinical practice. PMID- 12900268 TI - The year(s) of the contrast agent - micro-MRI in the new millennium. AB - The beginning of the new millennium has been a dynamic time for the field of magnetic resonance imaging (MRI). Exciting recent advances have been made at all levels of imaging, ranging from the visualization of single cells to rodents, birds and the human brain. Many of these techniques employ contrast agents to visualize the movement or activity of cells or organs in vivo; examples of this include the observation of stem cell migration, the tracking of labeled T cells, and the visualization of the events of gastrulation in developing Xenopus embryos. Other advances include improved techniques for elucidating white matter tracts in brain by either monitoring the diffusion of water along the tracts or tracing active neuronal tracts in vivo with Mn(2+) ions. Imaging of the immune system presents two dramatically different challenges: imaging most if not all of the body to follow cell trafficking, and imaging at cellular resolution to follow key intercellular and intracellular events. PMID- 12900270 TI - Recognition of microbial infection by Toll-like receptors. AB - The Toll-like receptors (TLRs) of the innate immune system detect host invasion by pathogens and initiate immune responses. All of the TLRs use the adaptor MyD88 to transduce a signal; however, two newly identified signaling molecules, TIRAP and TRIF, interact with a subset of the TLRs, suggesting a signaling specificity that may be relevant to the type of infection. Activation of the TRIF pathway, for example, leads to the production of antiviral gene products via the transcription factor, IRF3. In vivo experiments in TLR-deficient mice underscore the importance of TLRs in overcoming infection. PMID- 12900271 TI - Toll-like receptors and innate antiviral responses. AB - Toll-like receptors (TLRs) have a unique and important role in detecting the presence of pathogenic infection. TLRs can recognize conserved structures from a wide variety of microorganisms, such as bacteria, mycobacteria, spirochetes and yeast. However, they are generally not thought to play a major role in viral infection. Several reports have now identified distinct viral ligands for the TLRs, and evidence is accumulating for a functional role of the TLRs in mediating antiviral effector mechanisms. PMID- 12900272 TI - Innate sensors for Gram-positive bacteria. AB - More than half of invasive bacterial infections are Gram-positive in origin. This class of bacteria has neither endotoxins nor an outer membrane, yet it generates some of the most powerful inflammatory responses known in medicine. Some recent seminal studies go a long way toward settling the controversies that surround the process by which Gram-positive bacterial surfaces trigger the human immune system. Although the components of the cell wall are now chemically defined in exquisite detail and the interaction with the toll-like receptor 2 pathway has been discovered, it is only very recently that definitive studies combining these advanced biochemical and cell biological tools have been carried out. It is these breakthrough studies that have finally confirmed the paradigm of innate sensors for Gram-positive bacteria. PMID- 12900273 TI - The role of dendritic cell subsets in immunity to viruses. AB - Dendritic cells orchestrate the adaptive immune response. As well as presenting MHC-restricted antigen for T-cell activation, they provide all the co-receptor signals required for full T-cell priming. As a consequence, they play a central role in the immune response to infections caused by many pathogenic agents, including viruses. In recent times, it has become apparent that dendritic cells represent a particularly heterogeneous population with individual subsets playing specialized roles in response to infection. PMID- 12900274 TI - Shaping the immune response to parasites: role of dendritic cells. AB - Parasites represent a diverse group of pathogens that often trigger highly polarized immune responses that become tightly regulated during chronic infection. Recent studies have implicated the parasite-dendritic-cell interaction as a key determinant of the host response to these eukaryotic invaders. Dendritic cells appear to be pivotal in the initiation of cellular immunity against parasites as well as in directing Th1/Th2 effector choice. Moreover, there is increasing evidence that parasites regulate dendritic-cell function for the purposes of evading host immunity. This regulation also benefits parasites by protecting their host niche from the potentially lethal consequences of an uncontrolled inflammatory response. PMID- 12900275 TI - Cell-mediated immunity to respiratory virus infections. AB - The mucosal surfaces of the lungs pose tremendous problems for an immune system charged with maintaining a sterile pulmonary environment. Despite these problems, the immune system is effective at controlling most pulmonary infections. Over the past few years significant progress has been made in our understanding of how adaptive (humoral and cellular) immunity is able to control infections in the respiratory tract. Recent advances include the identification of effector memory T-cell populations in the lungs and an appreciation for the role of cytokines in regulating memory T-cell pools. PMID- 12900276 TI - Quantitative studies of CD8+ T-cell responses during microbial infection. AB - MHC class I tetramer staining, intracellular cytokine staining and ELISPOT assays have made it possible to quantify CD8(+) T-cell responses precisely during and following viral and bacterial infection. Although these quantitative methods are by now familiar and trusted components of the immunologist's toolbox, their application to models of microbial infection continues to provide surprising insights into mammalian adaptive immunity. In the past year there have been many exciting new findings on CD8(+) T-cell priming, expansion and memory formation in response to microbial infection. PMID- 12900277 TI - Subversion of immune responses by hepatitis C virus: immunomodulatory strategies beyond evasion? AB - Hepatitis C virus (HCV) is an important human pathogen that causes mild to severe liver disease worldwide. This positive-strand RNA virus is remarkably efficient at establishing persistent infection. In order for a non-cytopathic virus such as HCV to persist, the virus must escape immune recognition or inhibit the host immune responses. Immune escape via mutations in antigenic sites may occur under selective pressure during B-cell or T-cell responses to HCV infection, and may serve as a mechanism for the establishment HCV persistence. In addition to antigenic variation, HCV is able to subvert the host immune response by encoding specific viral gene product(s). An understanding of the mechanisms behind HCV persistence will provide a basis for the rational design of vaccines and novel therapeutic agents targeting human HCV infection. PMID- 12900278 TI - Immune evasion by Mycobacterium tuberculosis: living with the enemy. AB - Mycobacterium tuberculosis is successful as a pathogen because of its ability to persist in an immunocompetent host. This bacterium lives within the macrophage, a cell whose function is the elimination of microbes. Recent advances have improved our understanding of how M. tuberculosis evades two major antimicrobial mechanisms of macrophages: phagolysosome fusion and the production of toxic reactive nitrogen intermediates. M. tuberculosis also modulates antigen presentation to prevent the detection of infected macrophages by CD4(+) T cells. PMID- 12900279 TI - Vaccines for parasitic and bacterial diseases. AB - The first decade of the millennium should mark the beginning of a new era in vaccine development, reaping the rewards of advances in genome characterization, antigen identification, understanding the molecular bases of protective immune responses, and adjuvant design and development. Advances in all of these areas have culminated in vaccine candidates entering clinical testing. These include vaccines against two of humankind's oldest and deadliest diseases, tuberculosis and malaria. Several vaccine candidates for each of these diseases will be tested in humans during the next few years. A candidate vaccine for leishmaniasis, an infection that has taught us much about T-cell regulation of protection and disease in animal models, has been developed and is now in the clinic. There are indications both in animal models and in patients that vaccines may be used not only to protect but also to treat leishmania infections. PMID- 12900280 TI - Epitope-based vaccines: an update on epitope identification, vaccine design and delivery. AB - The basic premise of the epitope-based approach to vaccine development is that, in certain cases, the responses induced by the natural immunogen are not optimal, and can be improved upon by isolation or optimization of specific components of the response. For example, immunodominance is a key factor limiting the type and breadth of adaptive immunity. Recent advances in understanding the mechanisms of immunodominance thus represent an opportunity to further develop the epitope based approach. PMID- 12900281 TI - DNA vaccination: the answer to stable, protective T-cell memory? AB - Distinct populations of both CD4(+) and CD8(+) memory T cells have been identified on the basis of their location (lymphoid versus non-lymphoid tissues) and function. There have been several recent advances in our understanding of the factors that govern the generation and maintenance of T-cell memory in vivo. Utilising this knowledge could contribute to the refinement of DNA vaccine technology to induce more robust and longer-lasting protective T-cell based immunity. PMID- 12900282 TI - 11C-labeled stilbene derivatives as Abeta-aggregate-specific PET imaging agents for Alzheimer's disease. AB - A series of stilbene derivatives as potential diagnostic imaging agents targeting amyloid plaques in Alzheimer's disease (AD) were synthesized and evaluated. The syntheses of the stilbenes were successfully achieved by a simple Wadsworth Emmons reaction between diethyl (4-nitrobenzyl)phosphonate and 4 methoxybenzaldehyde. 4-N,N-dimethylamino-4'-methyoxy and the corresponding 4-N monomethylamino-, 4'-hydroxy stilbenes showed good binding affinities towards Abeta aggregates in vitro (K(i) < 10 nM). The (11)C labeled 4-N-methylamino-4' hydroxystilbene, [(11)C]4, was prepared by (11)C methylation of 4-amino-4' hydroxystilbene. The [(11)C]4 displayed a moderate lipophilicity (log P = 2.36), and showed a very good brain penetration and washout from normal rat brain after an iv injection. In vitro autoradiography of transgenic AD mouse brain sections showed a high specific labeling of beta-amyloid plaques, whereas the control sections showed no binding. Taken together the data suggest that a relatively simple stilbene derivative, [(11)C]4, N-[(11)C]methylamino-4'-hydroxystilbene, may be useful as a positron emission tomography (PET) imaging agent for mapping Abeta plaques in the brain of patients with Alzheimer's disease. PMID- 12900283 TI - Dimethylamino-fluorenes: ligands for detecting beta-amyloid plaques in the brain. AB - Formation of beta-amyloid (Abeta) plaques in the brain is a major contributing factor in the pathogenesis of Alzheimer's disease (AD). Detection of Abeta plaques in the brain will be potentially useful in early diagnosis and monitoring the progression of the disease. A series of novel Abeta aggregate-specific ligands based on fluorenes, which are simple and rigid tricyclic molecules, are synthesized and characterized. Starting with 2- or 3-aminofluorenes, 1a-1f, the amino group was converted to the N,N-dimethylamino group (2a-2f) in excellent yield. It was found that 7-iodo-2-N,N-dimethylaminofluorene (2f) showed an extremely high binding affinity to preformed Abeta40 aggregates (K(i) = 0.9 nM). In vitro autoradiography study using brain sections obtained from transgenic mice (Tg2576) with [(125)I]2f showed exquisitely high specific binding to Abeta plaques. The same section also displayed an equivalent labeling when stained by Thioflavin-S, a commonly used fluorescent dye for Abeta plaques. When [(125)I]2f was injected intravenously into normal mice, it exhibited an excellent brain uptake. Taken together the data suggest that [(125)I]2f may be useful as an in vivo imaging agent to detect Abeta plaques in the brain. PMID- 12900284 TI - Synthesis and evaluation of novel bifunctional chelating agents based on 1,4,7,10 tetraazacyclododecane-N,N',N",N"'-tetraacetic acid for radiolabeling proteins. AB - Detailed synthesis of the bifunctional chelating agents 2-methyl-6-(p isothiocyanatobenzyl)-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (1B4M-DOTA) and 2-(p-isothiocyanatobenzyl)-5, 6-cyclohexano-1, 4, 7, 10 tetraazacyclododecane-1, 4, 7, 10-tetraacetate (CHX-DOTA) are reported. These chelating agents were compared to 2-(p-isothiocyanatobenzyl)-1, 4, 7, 10 tetraazacyclododecane-1, 4, 7, 10-tetraacetic acid (C-DOTA) and 1, 4, 7, 10 Tetraaza-N-(1-carboxy-3-(4-nitrophenyl)propyl)-N', N", N"'-tris(acetic acid) cyclododecane (PA-DOTA) as their (177)Lu radiolabeled conjugates with Herceptin. In vitro stability of the immunoconjugates radiolabeled with (177)Lu was assessed by serum stability studies. The in vivo stability of the radiolabeled immunoconjugates and their targeting characteristics were determined by biodistribution studies in LS-174T xenograft tumor-bearing mice. Relative radiolabeling rates and efficiencies were determined for all four immunoconjugates. Insertion of the 1B4M moiety into the DOTA backbone increases radiometal chelation rate and provides complex stability comparable to C-DOTA and PA-DOTA while the CHX-DOTA appears to not form as stable a (177)Lu complex while exhibiting a substantial increase in formation rate. The 1B4M-DOTAmay have potential for radioimmunotherapy applications. PMID- 12900285 TI - In vitro and in vivo assessment of 99mTc-UBI specificity for bacteria. AB - Technetium-99m labeled ubiquicidin peptide 29-41 ((99m)Tc-UBI) is a cationic human antimicrobial peptide fragment that has been shown to bind bacteria in vitro and accumulates at sites of infection in experimental animals. To help determine if (99m)Tc-UBI is bound to the bacterial cell envelope by a simple nonspecific electrostatic interaction, a comparative study of the in vitro binding of (99m)Tc-UBI and two different (99m)Tc labeled cationic peptides ((99m)Tc-Tat-1-Scr and (99m)Tc-Tat-2-Scr) to bacteria and to two tumor cell line (LS174T and ACHN) was performed. The in vivo specificity of (99m)Tc-UBI for infection in mice was also evaluated using dual labels in the same animal and comparing the target/non-target ratio for (67)Ga-citrate and (99m)Tc-UBI at sites of induced infection and sterile inflammation. Under conditions of this study, the in vitro binding of (99m)Tc-UBI, (99m)Tc-Tat-1-Scr and (99m)Tc-Tat-2-Scr to S. aureus was 35, 78 and 87% respectively. While the binding of (99m)Tc-Tat-1-Scr and (99m)Tc-Tat-2-Scr was 37 and 33% to colon tumor cells (LS174T) and 39 and 41% to renal tumor cells (ACHN) respectively, the binding of (99m)Tc-UBI to both cell types was much lower at less than 4%. In vivo studies revealed that there is a significant difference (p < 0.05) in the radioactive accumulation of (99m)Tc-UBI between the sites of infection and inflammation compared to (67)Ga-citrate. Thus, (99m)Tc-UBI showed an average infection/inflammation ratio of 2.08 +/- 0.49 compared to 1.14 +/- 0.45 for (67)Ga-citrate. In conclusion, the in vitro and in vivo results provide evidence that a specific mechanism is responsible of the (99m)Tc-UBI bacterial intracellular accumulation. PMID- 12900286 TI - Lys and Arg in UBI: a specific site for a stable Tc-99m complex? AB - The aim of this study was to help establish if ubiquicidin peptide 29-41 fragment (UBI) contains a specific site for 99mTc labeling by a new direct method under alkaline conditions. Since this peptide does not have cysteine residues, it is possible that neighboring arginine and lysine in the peptide amino acid sequence (Thr-Gly-Arg-Ala-Lys-Arg-Arg-Met-Gln-Tyr-Asn-Arg-Arg) could be a specific coordination site to form a stable 99mTc-UBI complex. Following direct labeling, the in vitro stability of 99mTc-UBI was compared to UBI radiolabeled by one indirect method using HYNIC/tricine and HYNIC/tricine/EDDA. Radiochemical purity of 99mTc-UBI averaged 97% compared to 88% for 99mTc-HYNIC-UBI/tricine and 98% for 99mTc-HYNIC-UBI/tricine/EDDA. Both 99mTc-HYNIC-UBI (tricine or EDDA) and 99mTc UBI showed stability in human serum and solutions of cysteine. 99mTc-UBI radiochemical purity 24 h after dilution in 0.9% NaCl was greater than 90% at pH 9 and greater than 95% at pH 6.5. Under one set of experimental conditions, in vitro binding to bacteria of 99mTc-UBI was 35% and identical to that of 99mTc HYNIC-UBI/tricine and 99mTc-HYNIC-UBI/tricine/EDDA at 32% and 31% respectively. The biodistribution of 99mTc-UBI in mice showed a rapid renal clearance. To help identify the site(s) of 99mTc binding following direct labeling, molecular mechanics and quantum-mechanical calculations were performed which showed that the amine groups of Arg(7) and Lys are the most probable site. The calculations show that these groups can form a square pyramid with two water molecules for the Tc cation (dxysp(3)). It will be necessary to isolate and characterize the 99Tc(V)(O)-UBI.(H2O)n complex to confirm these results. PMID- 12900287 TI - Synthesis of dendrimer-based biotin radiopharmaceuticals to enhance whole-body clearance. AB - To synthesize a biotin radiopharmaceutical that clears rapidly, dendrimer was used as a carrier and conjugated with succinimidyl 3-[(125)I]iodobenzoate and tetrafluorophenyl norbiotinamidosuccinate. Then, succinic anhydride was used to reduce its pI. In mice, the non-succinylated product showed high liver (67% ID/g) and kidney (44% ID/g) uptakes and whole-body retention (94% ID) at 20 min that persisted for 12 hr. The corresponding organ uptakes (22% and 11% ID/g) and the whole-body retention (47% ID) were drastically reduced by succinylation (p<0.0001). Lysine co-injection further lowered renal uptake. PMID- 12900288 TI - To predict response chemotherapy using technetium-99m tetrofosmin chest images in patients with untreated small cell lung cancer and compare with p-glycoprotein, multidrug resistance related protein-1, and lung resistance-related protein expression. AB - Our preliminary studies found technetium-99m tetrofosmin (Tc- TF) chest imaging was related to Pgp or MRP1 expression and successfully predict chemotherapy response and in SCLC in human. However, there was no published literature to study relationship of Tc-TF chest images and LRP expression in SCLC patients. Therefore, the aim of this study was to investigate the relationships among Tc- TF accumulation in untreated small cell lung cancer (SCLC), the expression of P glycoprotein (Pgp), multidrug resistance related protein-1 (MRP1), and lung resistance-related protein (LRP), as well as the response to chemotherapy in patients with untreated SCLC. Thirty patients with SCLC were studied with chest images 15 to 30 minutes after intravenous injection of Tc-TF before chemotherapeutic induction. Tumor-to-background (T/B) ratios were obtained on the static and plantar Tc-TF chest images. The response to chemotherapy was evaluated upon completion of chemotherapy by clinical and radiological methods. These patients were separated into 15 patients with good response and 15 patients with poor response. No significant differences of prognostic factors (Karnofsky performance status, tumor size, or tumor stage) were found between the patients with good and poor responses. Immunohistochemical analyses were performed on multiple nonconsecutive sections of biopsy specimens to detect Pgp, MRP1, and LRP expression. The difference in T/B ratios on the Tc-TF chest images of the patients with good versus poor response was significant. The differences in T/B ratios of the patients with positive versus negative Pgp expression and with positive versus negative MRP1 expression were significant. The difference in T/B ratios of the patients with positive versus negative LRP expression was not significant. We concluded that Tc-TF chest images could accurately predict chemotherapy response of patients with SCLC. In addition, The Tc-TF tumor uptake was related to Pgp or MRP1 but not LPR expression in SCLC. PMID- 12900289 TI - Technetium-99m scintigraphy to visualize T-cell homing in vivo: a preclinical study. AB - The knowledge of lymphocyte distribution is very usefulness in monitoring therapeutic treatments. We present here a method employed in clinical practice, the scintigraphy, to study in the rat the physiologic lymphocyte traffic. Rat T cells labeled with 99mTc were injected in syngeneic animals, and their fate was studied by serial scintigraphic scanning. Sorted naive CD4+ CD45RC(bright) T cells homed to lymphoid organs and accumulated in spleen. CD4+ CD45RC(dim) memory lymphocytes first reached the liver and the lungs and recirculated. The results obtained by using the scintigraphic method to in vivo study the lymphocyte homing in rats are comparable to those obtained with previously used experimental methods. We consider the scintigraphic method a useful tool to in vivo track lymphocytes and to address therapeutic treatment in men. PMID- 12900290 TI - Imaging of striatal dopamine transporters in rat brain with single pinhole SPECT and co-aligned MRI is highly reproducible. AB - A recently developed pinhole high-resolution SPECT system was used to measure striatal to non-specific binding ratios in rats (n = 9), after injection of the dopamine transporter ligand (123)I-FP-CIT, and to assess its test/retest reproducibility. For co-alignment purposes, the rat brain was imaged on a 1.5 Tesla clinical MRI scanner using a specially developed surface coil. The SPECT images showed clear striatal uptake. On the MR images, cerebral and extra cerebral structures could be easily delineated. The mean striatal to non-specific [(123)I]FP-CIT binding ratios of the test/retest studies were 1.7 +/- 0.2 and 1.6 +/- 0.2, respectively. The test/retest variability was approximately 9%. We conclude that the assessment of striatal [(123)I]FP-CIT binding ratios in rats is highly reproducible. PMID- 12900291 TI - Evaluation of [18F]FHPG as PET tracer for HSVtk gene expression. AB - In rats, the relationship between [(18)F]FHPG accumulation and HSVtk expression was studied with PET and autoradiography. [(18)F]FHPG distribution closely corresponded with HSVtk immunohistochemical staining. ROI analysis of tracer uptake 2 hours p.i. and Patlak graphical analysis were applied to quantify the PET data. For both analysis methods, the [(18)F]FHPG PET signal correlated well with the fraction of HSVtk expressing cells implanted, but showed a plateau when plotted against HSVtk protein levels. This might be due to rate limiting [(18)F]FHPG membrane transport. PMID- 12900292 TI - Evaluation of radioiodinated 8-Cyclopentyl-3-[(E)-3-iodoprop-2-en-1-yl]-1 propylxanthine ([*I]CPIPX) as a new potential A1 adenosine receptor antagonist for SPECT. AB - 8-Cyclopentyl-3-[(E)-3-[(131)I]iodoprop-2-en-1-yl]-1-propylxanthine (2*) was generated by iododestannylation of the tributyl-stannyl-precursor with [(131)I]NaI and chloramine T. The radiochemical yield of 2* was 82 +/- 4%, and the purity exceeded 98%. The specific activity was 33 +/- 19 GBq/micromol. Affinities for rat, pig and human A(1) adenosine receptors (A(1)ARs) were in the low nanomolar range, but poor selectivity for the human A(1)AR over the A(2A)AR was found. Additionally, in vitro and ex vivo autoradiographic studies revealed high unspecific binding which makes this ligand unsuitable for SPECT imaging. PMID- 12900293 TI - Demonstration of high monoaminoxidase-A levels in neuroendocrine gastroenteropancreatic tumors in vitro and in vivo-tumor visualization using positron emission tomography with 11C-harmine. AB - BACKGROUND AND AIMS: A majority of neuroendocrine gastroenteropancreatic (GEP) tumors can be detected by conventional radiological methods and scintigraphic techniques. Still there are problems to visualize small tumor lesions and non functioning tumors. The aim of this study was to investigate some of the monoamine processing pathways of neuroendocrine GEP-tumors and try to find a new tracer substance for in vivo characterization and visualization by Positron Emission Tomography (PET). SUBJECTS AND METHODS: Autoradiography of tumor sections from 8 midgut carcinoids (MGC) and 8 endocrine pancreatic tumors (EPT) was performed with (11)C-labeled tracers for serotonin and dopamine transporters, serotonin HT2A-, dopamine D1- and muscarinic receptors and for monoamine oxidase A (MAO-A). The in vitro results initiated PET studies with (11)C-Harmine in 4 patients with MGC and 7 patients with EPT (one insulinoma, two glucagonomas and four non-functioning EPT). RESULTS: The MAO-A-ligand Harmine expressed specific in vitro binding of 87 +/-21% for MGC and 125 +/- 50% for EPT, compared to reference tissue (rat brain, 100%). All other substances showed relatively low specific binding. (11)C-harmine-PET could visualize tumors in all patients. The mean standardized uptake value (SUV) for MGC was 7.5 +/- 3.9 and for EPT 12.9 +/- 2.7, whereas the SUV of normal liver, intestine and pancreas were 3.1 +/- 0.5, 3.4 +/- 1.2 and 8.9 +/- 3.0 respectively. CONCLUSIONS: This study demonstrates in vitro and in vivo that neuroendocrine GEP-tumors are characterized by a high MAO A-expression, thereby adding to the similarities of neuronal and neuroendocrine tissue. It also indicates a possible application for (11)C-harmine as a new PET tracer for neuroendocrine GEP-tumors with the potential to visualize also non functioning EPT's. PMID- 12900294 TI - The value of referring to recently introduced antipsychotics as "second generation". PMID- 12900295 TI - Mental health. AB - OBJECTIVE: Only in the last 30 years has psychiatry begun to develop empirical approaches to conceptualizing and assessing positive mental health. Six models of mental health are reviewed here. METHOD: The author points out pitfalls in research on mental health, e.g., equating average with healthy, failing to distinguish trait from state, overlooking cultural norms, and conversely, blindly accepting the culture's values. He describes the six models and provides history and research needs for each. RESULTS: The first model, being "above normal," is epitomized by DSM-IV's axis V, the Global Assessment of Functioning Scale. High scores represent "superior functioning in a wide range of activities, life's problems never seem to get out of hand, is sought out by others because of his or her many positive qualities." The goal of the second model, positive psychology, is intervention to maximize positive qualities, such as self-efficacy. Maturity and Erikson's four developmental tasks (identity, intimacy, generativity, integrity) are the basis of the third model. The author adds two other tasks: career consolidation and "keeper of the meaning." The fourth model is emotional or social intelligence, the ability to read other people's emotions. Surprisingly, subjective well-being, the fifth model, is as much a characteristic of temperament as of a benign environment. The last model, resilience, is epitomized by DSM-IV's Defense Function Scale, which categorizes coping mechanisms in terms of adaptational value. CONCLUSIONS: As with the blind men and the elephant, each model describes only some aspects of mental health. Further research may reveal the contribution of each. PMID- 12900296 TI - Examining the effects of prevention programs on the incidence of new cases of mental disorders: the lack of statistical power. AB - OBJECTIVE: In the past few decades about 1,000 controlled studies have examined the effects of mental health prevention programs, but few studies have examined the effects of such programs on the incidence of new cases of mental disorders defined according to diagnostic criteria. A major reason why so few studies have examined this important question is that very large numbers of subjects are needed to provide sufficient statistical power for these studies. In this article this power problem is explored. METHOD: Power calculations are presented for studies examining universal prevention (aimed at the general population regardless of risk status), selective prevention (aimed at high-risk groups), and indicated prevention (aimed at subjects who have some symptoms of a disorder without meeting full diagnostic criteria). RESULTS: Studies examining universal prevention are hardly feasible, as the number of subjects required amounts to tens of thousands at least. Research examining selective prevention is more feasible, but the number of subjects needed for these studies is still very high. Studies of indicated prevention are possible. Three major studies of indicated prevention examining the effects on the incidence of new cases of mental disorders are described. CONCLUSIONS: There are several strategies for increasing statistical power in prevention studies: 1) focus on high-incidence groups (by concentrating on indicated prevention, by targeting high-risk groups with multiple risk factors, by targeting groups with multiple disorders), 2) strengthen the effects of prevention programs, and 3) make more efficient use of other trials through cumulative meta-analyses. PMID- 12900297 TI - Schizophrenia, IV: neuregulin-1 in the human brain. PMID- 12900300 TI - Comparative efficacy and safety of atypical and conventional antipsychotic drugs in first-episode psychosis: a randomized, double-blind trial of olanzapine versus haloperidol. AB - OBJECTIVE: Few long-term studies have compared the efficacy and safety of typical and atypical antipsychotic medications directly in patients with a first episode of psychosis who met the criteria for schizophrenia or a related psychotic disorder. This study compared the acute and long-term effectiveness of haloperidol with that of olanzapine in patients with first-episode psychosis in a large, controlled clinical trial. METHOD: Patients with first-episode psychosis (N=263) were randomly assigned under double-blind conditions to receive haloperidol or olanzapine and were followed for up to 104 weeks. Domains measured included psychopathology, psychosocial variables, neurocognitive functioning, and brain morphology and metabolism. This report presents data from clinical measures of treatment response and safety data from the 12-week acute treatment phase. RESULTS: Haloperidol and olanzapine were associated with substantial and comparable baseline-to-endpoint reductions in symptom severity, which did not differ significantly in last-observation-carried-forward analyses. However, in a mixed-model analysis, olanzapine-treated subjects had significantly greater decreases in symptom severity as measured by the Positive and Negative Syndrome Scale total score and negative and general scales and by the Montgomery-Asberg Depression Rating Scale but not as measured by the Positive and Negative Syndrome Scale positive scale and by the Clinical Global Impression severity rating. Olanzapine-treated patients experienced a lower rate of treatment-emergent parkinsonism and akathisia but had significantly more weight gain, compared with the haloperidol-treated patients. Overall, significantly more olanzapine-treated subjects than haloperidol-treated subjects completed the 12-week acute phase of the study (67% versus 54%). CONCLUSIONS: As expected on the basis of previous studies, both olanzapine and haloperidol were effective in the acute reduction of psychopathological symptoms in this group of patients with first-episode psychosis. However, olanzapine had several relative advantages in therapeutic response. Although the nature of adverse events differed between the two agents, retention in the study was greater with olanzapine. Retention in treatment is important in this patient population, given their risk of relapse. Longer-term results are needed to determine whether treatment with atypical antipsychotics results in superior outcomes for a first episode of schizophrenia. PMID- 12900301 TI - Maintenance treatment of schizophrenia with risperidone or haloperidol: 2-year outcomes. AB - OBJECTIVE: Most controlled studies comparing second-generation and conventional antipsychotics have focused on the acute treatment of schizophrenia. The authors compared symptom outcomes, side effects, and social adjustment in stable schizophrenia outpatients who received 2 years of maintenance treatment with risperidone or haloperidol. METHOD: This was a 2-year, randomized, double-blind comparison of 6 mg of risperidone versus haloperidol in 63 patients with stabilized DSM-IV schizophrenia. Study patients also received 15 months of standard behavioral skills training or enhanced training with a case manager who promoted patients' use of their skills in the community. RESULTS: The risk of psychotic exacerbations and the risk of leaving the study were similar for both drug treatment groups. However, patients who received both risperidone and the enhanced community-based skills training were more likely to remain in the study than those in the other treatment groups. Patients demonstrated significant improvement in score on the Brief Psychiatric Rating Scale over time with both medications. There were no between-group differences in cluster scores for thought disturbance, hostile-suspiciousness, and withdrawal-retardation. A significant between-group difference favoring risperidone was found for the anxious-depression cluster. Risperidone resulted in significantly greater reductions in tremor and akathisia and greater improvements in most items on the SCL-90-R. CONCLUSIONS: When compared with patients given a low dose of haloperidol, risperidone-treated patients experienced similar improvements in positive and negative symptoms and similar risks of psychotic exacerbations. However, risperidone-treated patients appeared to feel subjectively better, as indicated by less anxiety and depression and fewer extrapyramidal side effects. PMID- 12900302 TI - Is regionally selective D2/D3 dopamine occupancy sufficient for atypical antipsychotic effect? an in vivo quantitative [123I]epidepride SPET study of amisulpride-treated patients. AB - OBJECTIVE: Atypical antipsychotic drug treatment is clinically effective with a low risk of extrapyramidal symptoms. Explanations for the mechanism underlying this beneficial therapeutic profile of atypical over typical antipsychotic agents include 1) simultaneous antagonism of dopamine D(2) and serotonin 5-HT(2A) receptors or 2) selective action at limbic cortical dopamine D(2)-like receptors with modest striatal D(2) receptor occupancy. Amisulpride is an atypical antipsychotic drug with selective affinity for D(2)/D(3) dopamine receptors and provides a useful pharmacological model for examining these hypotheses. The authors' goal was to evaluate whether treatment with amisulpride results in "limbic selective" D(2)/D(3) receptor blockade in vivo. METHOD: Five hours of dynamic single photon emission tomography data were acquired after injection of [(123)I]epidepride (approximately 150 MBq). Kinetic modeling was performed by using the simplified reference region model to obtain binding potential values. Estimates of receptor occupancy were made relative to a healthy volunteer comparison group (N=6). RESULTS: Eight amisulpride-treated patients (mean dose=406 mg/day) showed moderate levels of D(2)/D(3) receptor occupancy in the striatum (56%), and significantly higher levels were seen in the thalamus (78%) and temporal cortex (82%). CONCLUSIONS: Treatment with amisulpride results in a similar pattern of limbic cortical over striatal D(2)/D(3) receptor blockade to that of other atypical antipsychotic drugs. This finding suggests that modest striatal D(2) receptor occupancy and preferential occupancy of limbic cortical dopamine D(2)/D(3) receptors may be sufficient to explain the therapeutic efficacy and low extrapyramidal symptom profile of atypical antipsychotic drugs, without the need for 5-HT(2A) receptor antagonism. PMID- 12900303 TI - The relationship of clozapine and haloperidol treatment response to prefrontal, hippocampal, and caudate brain volumes. AB - OBJECTIVE: The study was designed to assess the predictive relationship between brain structure volume and positive and negative symptom response to clozapine and haloperidol. METHOD: Partially responsive outpatients with schizophrenia who participated in a 10-week, parallel-group, double-blind comparison of clozapine and haloperidol and who had an available magnetic resonance imaging scan were included in the current study. Prefrontal gray and white matter, hippocampal, and caudate volumes were manually measured. The Scale for the Assessment of Negative Symptoms (SANS) and the Brief Psychiatric Rating Scale (BPRS) were used to assess symptom changes. The Simpson-Angus Rating Scale was used to assess extrapyramidal symptoms. RESULTS: Twenty-two patients randomly assigned to clozapine and 23 patients assigned to haloperidol met study entry criteria. There were significant interactions between treatment and right prefrontal gray matter volume for BPRS total score and SANS total score. There were no significant treatment-by-brain structure interactions for BPRS positive symptom items. Right prefrontal gray matter volume was also related to differential treatment effects for the BPRS subscales of anxiety/depression and hostility and the Simpson-Angus Rating Scale akathisia item. CONCLUSIONS: These results suggest that there is a differential interaction among clozapine and haloperidol, brain structure, and treatment response. Partially responsive patients with larger brain volumes may be more likely to experience the benefits of clozapine treatment, but they may be more vulnerable to side effects and experience a subsequent worsening of their symptoms when treated with haloperidol. PMID- 12900304 TI - Transfer of olanzapine into breast milk, calculation of infant drug dose, and effect on breast-fed infants. AB - OBJECTIVE: This study characterized infant drug doses and breast-milk-to-plasma area-under-the-curve ratios for olanzapine and determined plasma concentrations and effects of this drug on breast-feeding infants. METHOD: Seven mother-infant nursing pairs were studied. Olanzapine was measured in plasma and milk with high performance liquid chromatography over a dose interval (for six patients) or at a single time after dose ingestion (for one patient) at steady state. Infant drug exposure was estimated as the product of an assumed milk production rate and average drug concentration in milk, normalized to body weight, and expressed as a percentage of maternal drug dose, normalized to body weight. RESULTS: The median infant dose of olanzapine ingested through milk was 1.02% of the maternal dose; the median milk-to-plasma area-under-the-curve ratio was 0.38 for the six patients with data collected over the dose interval. Corresponding values in the patient with single-point data were 1.13% and 0.75. Olanzapine was not detected in the plasma of the six infants with an evaluable plasma sample. All of the infants were healthy and experienced no side effects. CONCLUSIONS: Breast-fed infants were exposed to a calculated olanzapine dose of approximately 1%-well below the 10% notional level of concern. In infant plasma, olanzapine was below the detection limit; there were no adverse effects on the infants. These data support the use of olanzapine during breast-feeding. However, the authors recommend that breast-fed infants be monitored closely and the decision to breast feed be made after individual risk-benefit analysis. PMID- 12900305 TI - Are benzodiazepines still the medication of choice for patients with panic disorder with or without agoraphobia? AB - OBJECTIVE: Recently, pharmacological treatment guidelines for panic disorder have changed as newer treatment options have become available. The authors examined how the use of psychotropic drugs has shifted over the course of 10 years to determine if prescribing patterns have changed to reflect these revised treatment guidelines. METHOD: A total of 443 patients with panic disorder were enrolled in the Harvard/Brown Anxiety Research Project, a prospective longitudinal study of anxiety disorders. These patients were interviewed over the course of 10 years to examine their use of psychotropic medications. RESULTS: Despite efforts aimed at increasing the use of selective serotonin reuptake inhibitors (SSRIs) in patients with panic disorder (e.g., APA's practice guideline for panic disorder, Food and Drug Administration approval of particular SSRIs for the treatment of panic disorder), only a modest increase in their use was found. Treatment patterns for psychotropic drugs appear to have remained stable over the past decade, with benzodiazepines being the most commonly used medication for panic disorder. In comparison, SSRI use throughout the follow-up period has remained low. Patients using an SSRI did not have a more favorable clinical course than those using a benzodiazepine, nor were there significantly better rates of remission in patients using SSRIs and benzodiazepines concomitantly. CONCLUSIONS: These results highlight a gap between pharmacological treatment guidelines and actual delivery of care in that recommendations to use SSRIs to treat panic disorder are not being followed. Factors potentially associated with promoting and ignoring treatment recommendations are discussed. PMID- 12900306 TI - Enhanced salience and emotion recognition in Autism: a PET study. AB - OBJECTIVE: This study examined neural activation of facial stimuli in autism when the salience of emotional cues was increased by prosodic information. METHOD: Regional cerebral blood flow (rCBF) was measured while eight high-functioning men with autism and eight men without autism performed an emotion-recognition task in which facial emotion stimuli were matched with prosodic voices and a baseline gender-recognition task. RESULTS: Emotion processing in autistic subjects, compared to that in comparison subjects, resulted in lower rCBF in the inferior frontal and fusiform areas and higher rCBF in the right anterior temporal pole, the anterior cingulate, and the thalamus. CONCLUSIONS: Even with the enhanced emotional salience of facial stimuli, adults with autism showed lower activity in the fusiform cortex and differed from the comparison subjects in activation of other brain regions. The authors suggested that the recognition of emotion by adults with autism is achieved through recruitment of brain regions concerned with allocation of attention, sensory gating, the referencing of perceptual knowledge, and categorization. PMID- 12900307 TI - Proton magnetic resonance spectroscopy of bipolar disorder versus intermittent explosive disorder in children and adolescents. AB - OBJECTIVE: The diagnosis of bipolar disorder in juveniles is controversial. This study was designed to compare proton magnetic resonance spectroscopy ((1)H MRS) in patients with bipolar disorder or intermittent explosive disorder, two groups with symptomatic overlap but categorical distinction. Children with intermittent explosive disorder designate patients whose illness clinically resembles pediatric bipolar disorder but does not satisfy DSM-IV criteria for mania. Based on the authors' previous report of higher levels of (1)H MRS cingulate myo inositol/creatine in youngsters with bipolar disorder than in normal comparison subjects, they hypothesized that patients with bipolar disorder would have higher cingulate myo-inositol/creatine-phosphocreatine measurements than patients with intermittent explosive disorder and normal comparison subjects. METHOD: Myo inositol levels were measured with a 2x2x2 cm(3) voxel placed in the anterior cingulate for acquisition of (1)H MRS in 10 patients with bipolar disorder, 10 patients with intermittent explosive disorder, and 13 normal comparison subjects. N-Acetylaspartate, choline moieties, creatine-phosphocreatine, and glutamate glutamine metabolite levels were also measured. RESULTS: The patients with bipolar disorder showed significantly higher anterior cingulate myo inositol/creatine-phosphocreatine and myo-inositol (mmol/liter) levels than the patients with intermittent explosive disorder and the normal comparison subjects. No significant differences were found across groups for myo-inositol or other metabolites in the occipital cortex. CONCLUSIONS: These data provide evidence that differences in the concentration of myo-inositol (mmol/liter) in the anterior cingulate cortex in (1)H MRS may differentiate these two populations. Follow-up studies involving larger samples may conclusively estimate the biological specificity between pediatric bipolar disorder and other disorders, which overlap clinically. PMID- 12900308 TI - Relationship between multiple forms of childhood maltreatment and adult mental health in community respondents: results from the adverse childhood experiences study. AB - OBJECTIVE: This study examined the prevalence of a history of various combinations of childhood maltreatment types (physical abuse, sexual abuse, and witnessing of maternal battering) among adult members of a health maintenance organization (HMO) and explored the relationship with adult mental health of the combinations of types of childhood maltreatment and emotional abuse in the childhood family environment. METHOD: A total of 8,667 adult members of an HMO completed measures of childhood exposure to family dysfunction, which included items on physical and sexual abuse, witnessing of maternal battering, and emotional abuse in the childhood family environment. The adults' current mental health was assessed by using the mental health scale of the Medical Outcomes Study 36-item Short-Form Health Survey. RESULTS: The prevalences of sexual abuse, physical abuse, and witnessing of maternal violence were 21.6%, 20.6%, and 14.0%, respectively, when the maltreatment types were considered separately. Among respondents reporting any of the maltreatment types, 34.6% reported more than one type of maltreatment. Lower mean mental health scores were associated with higher numbers of abuse categories (mean=78.5, 75.5, 72.8, and 69.9 for respondents with no, one, two, and three abuse types, respectively). Both an emotionally abusive family environment and the interaction of an emotionally abusive family environment with the various maltreatment types had a significant effect on mental health scores. CONCLUSIONS: Childhood physical and sexual abuse, as well as witnessing of maternal battering, were common among the adult members of an HMO in this study. Among those reporting any maltreatment, more than one-third had experienced more than one type of maltreatment. A dose-response relation was found between the number of types of maltreatment reported and mental health scores. In addition, an emotionally abusive family environment accentuated the decrements in mental health scores. Future research examining the effects of childhood maltreatment on adult mental health should include assessments of a wide range of abusive experiences, as well as the family atmosphere in which they occur. PMID- 12900309 TI - Discriminative validity and clinical utility of an abuse-neglect interview for adolescents with conduct and substance use problems. AB - OBJECTIVE: Many youths with conduct and substance use problems experience abuse and neglect. A valid measure of the severity of abuse-neglect events could facilitate research and clinical care. The authors' goal was to examine the discriminative validity, clinical utility, and severity correlations of the Colorado Adolescent Rearing Inventory, a structured, 20-45-minute abuse-neglect interview in the assessment of adolescents with conduct and substance use problems. METHOD: Ninety-eight adolescent patients with conduct and substance use problems and 102 comparison subjects (about 40% of the subjects in each group were female) completed the Colorado Adolescent Rearing Inventory, the Diagnostic Interview Schedule for Children, and the Composite International Diagnostic Interview-Substance Abuse Module. RESULTS: Although the subjects were advised that reports of abuse-neglect would be communicated to child welfare agencies, nearly two-thirds of the patients with conduct and substance use problems endorsed items indicating probable abuse-neglect, compared with about a third of the comparison subjects, a highly significant difference. Clinicians judged that 68%-80% of the cases of the patients and comparison subjects who endorsed such items warranted reporting to child welfare agencies. Scores on the Colorado Adolescent Rearing Inventory correlated significantly with severity of substance involvement, conduct disorder, and major depression. Compared to males, significantly more females (including 15% of comparison females) reported sexual abuse. Patients were significantly more likely than comparison subjects to attribute adverse life effects to their reported abuse-neglect experiences. CONCLUSIONS: Colorado Adolescent Rearing Inventory scores revealed many cases of serious abuse-neglect, generated many reports to child welfare agencies, demonstrated discriminative validity, correlated with clinical measures, and reflected important gender differences. The Colorado Adolescent Rearing Inventory can contribute to clinical evaluation and research involving youths with conduct and substance use problems. PMID- 12900310 TI - Early risk factors for violence in Colombian adolescents. AB - OBJECTIVE: Violence and homicide are more prevalent in Colombia, South America, than in the United States, but the role of psychosocial factors in the violent behavior of Colombian adolescents remains unclear. The objective of the study was to identify personality, familial, peer, and ecological variables associated with violence in Colombian adolescents. METHOD: A survey of adolescents was conducted in 1995-1996. A standard self-report measure was adapted to ensure linguistic and cultural relevance. A total of 2,837 adolescents ages 12-17 years from various self-reported ethnic groups were randomly selected from the community in three Colombian cities: Bogota, Medellin, and Barranquilla. Eighty percent of eligible adolescents agreed to participate. Data were collected concerning the adolescent's personality attributes, family characteristics, peer characteristics, and ecological/cultural factors, including the availability of illicit drugs and the prevalence of violence in the community. The dependent variable was the adolescent's self-reported frequency of violent behavior. RESULTS: Violence directed at the adolescent and the adolescent's own drug use were both more highly correlated with the adolescent's violent behavior than were other risk factors. Significant risk factors of less importance included tolerance of deviance, peer drug use, peer deviance, and exposure to violence on television. CONCLUSIONS: The results supported a model in which violent behavior was correlated independently with a number of risk factors from several domains. The findings point to the use of specific intervention procedures for adolescents to prevent their own subsequent acts of violent behavior. PMID- 12900311 TI - Comparisons of problems reported by youths from seven countries. AB - OBJECTIVE: This study compared ratings for self-reported behavioral and emotional problems in adolescents from seven countries. METHOD: Youth Self-Report scores were analyzed for 7,137 adolescents ages 11-18 years from general population samples from Australia, China, Israel, Jamaica, the Netherlands, Turkey, and the United States. RESULTS: Comparisons of problems scores yielded small to medium effect sizes for cross-cultural variations. Youths from China and Jamaica had the highest and youths from Israel and Turkey had the lowest mean total problems scores. With cross-cultural consistency, girls scored higher for internalizing and lower for externalizing than boys. Cross-cultural correlations were high among the mean item scores. CONCLUSIONS: Empirically based assessment provided a robust method for assessing and comparing adolescents' self-reported problems. Self-reports thus supplemented empirically based assessments of parent-reported problems and offered a cost-effective way of identifying problems for which adolescents from diverse cultural backgrounds may need help. PMID- 12900312 TI - Peripubertal suicide attempts in offspring of suicide attempters with siblings concordant for suicidal behavior. AB - OBJECTIVE: The authors sought to determine 1) whether the risk for familial transmission of suicidal behavior is greater with increased family loading for suicide attempts, and 2) whether the transmission of suicidal behavior is mediated by impulsive aggression. METHOD: A reanalysis of a high-risk study compared the offspring of three mood disorder proband groups: suicide attempters with a sibling who also attempted suicide (N=19), suicide attempters whose siblings never made a suicide attempt (N=73), and nonsuicidal probands whose siblings also never engaged in suicidal behavior (N=73). Probands and offspring were assessed with respect to psychopathology, suicide attempt history, impulsive aggression, and exposure to familial adversity. RESULTS: Offspring of suicide attempters with siblings concordant for suicidal behavior showed a higher risk of suicide attempt than did offspring of nonsuicidal probands and had an earlier age at onset of suicidal behavior than offspring of suicide attempters with siblings discordant for suicidal behavior. Probands from sibling pairs concordant for suicidal behavior and their offspring reported greater lifetime impulsive aggression compared with each of the other two proband/offspring groups. In the offspring, impulsive aggression was the most powerful predictor of early age at first suicide attempt. CONCLUSIONS: Familial loading for suicide attempts may affect rates of transmission as well as age at onset of suicidal behavior, and its effect may be mediated by the familial transmission of impulsive aggression. PMID- 12900313 TI - Comorbidity of axis I and axis II disorders in patients who attempted suicide. AB - OBJECTIVE: The authors compared the characteristics of suicide attempters with and without comorbid psychiatric and personality disorders to identify factors that explain the high suicide risk associated with psychiatric comorbidity. METHOD: A representative group of 111 patients who had attempted suicide (72 female and 39 male) was assessed for psychiatric and personality disorders according to ICD-10 criteria. The characteristics of patients with both types of disorder were compared with those of patients without comorbid disorders. A semistructured interview schedule and standardized questionnaires were used to investigate patients' background characteristics, the circumstances of the suicide attempts, psychological characteristics, and outcome after 12-20 months. RESULTS: Comorbidity of psychiatric and personality disorders was present in 49 patients (44%). More patients with comorbid disorders had made previous suicide attempts (N=41 [84%] versus N=28 [45%]) and repeated attempts during the follow up period (N=25 [51%] versus N=9 [15%]). Differences in precipitants and motives for the index episode were also found: patients with comorbid disorders were more depressed and hopeless, reported more episodes of aggression, were more impulsive, and had lower self-esteem and poorer problem-solving skills. Differences in self-esteem and problem-solving skills distinguished between the groups in a stepwise discriminant function analysis. More of the patients with comorbid disorders reported not being loved by their parents and parental suicidal behavior. CONCLUSIONS: Suicide attempters with comorbid psychiatric and personality disorders show marked differences from those without both of these disorders. Comorbidity may contribute to greater suicide risk. Some of the characteristics of patients with comorbid disorders pose major clinical challenges that should be addressed in an effort to reduce suicide risk. PMID- 12900314 TI - Deliberate self-harm in a nonclinical population: prevalence and psychological correlates. AB - OBJECTIVE: Research on deliberate self-harm (intentionally injuring oneself without suicidal intent) has focused on clinical and forensic populations. Studying only these populations, which typically have serious psychopathology, may lead to inflated estimates of the association between self-harm and psychiatric disorder, as well as of the prevalence of deliberate self-harm. The present study investigated the prevalence and correlates of deliberate self-harm in a large group of nonclinical subjects. METHOD: Participants were 1,986 military recruits, 62% of whom were men, who were participating in a study of peer assessment of personality traits and pathology. Individuals who did and did not report a history of self-harm were compared on measures of personality and psychopathology. RESULTS: Approximately 4% of the participants reported a history of deliberate self-harm. Compared with participants without a history of deliberate self-harm, self-harmers scored higher on self- and peer-report measures of borderline, schizotypal, dependent, and avoidant personality disorder symptoms and reported more symptoms of anxiety and depression. Item-level analyses indicated that peers viewed self-harmers as having strange and intense emotions and a heightened sensitivity to interpersonal rejection. CONCLUSIONS: About one of every 25 members of a large group of relatively high-functioning nonclinical subjects reported a history of self-harm. Self-harmers had more symptoms of several personality disorders than non-self-harmers, and their performance across measures suggested that anxiety plays a prominent role in their psychopathology. Future research should investigate whether psychotherapies or psychiatric medications known to reduce symptoms of anxiety can be effective in treating deliberate self-harm. PMID- 12900315 TI - Psychopathology and comorbidity of psychiatric disorders in patients with kleptomania. AB - OBJECTIVE: This study compared patients with kleptomania, patients with alcohol abuse or dependence, and psychiatric patients without impulse-control disorders or substance-related disorders on several key psychopathological dimensions. In addition, the comorbidity of kleptomania with other psychiatric disorders was examined. METHOD: Eleven patients with kleptomania recruited over a cumulative 2 year period and 60 patients with alcohol abuse or dependence and 29 psychiatric comparison patients recruited over a consecutive 6-month period participated in structured clinical interviews to determine the presence of impulse-control and substance-related disorders and of other psychiatric disorders that were comorbid with kleptomania. Psychopathological dimensions were measured with the Barratt Impulsiveness Scale, the Sensation Seeking Scale, the Montgomery-Asberg Depression Rating Scale, and the anxiety and depression subscales of the Hospital Anxiety and Depression Scale. RESULTS: Significant group effects were found for the Barratt Impulsiveness Scale total and cognitive impulsivity scores, with the patients with kleptomania having higher impulsivity scores than the other groups. Significant group differences were found on the Sensation Seeking Scale total and disinhibition scores. No significant group effects were found for the mood and anxiety measures. Patients with kleptomania had high rates of comorbid psychiatric disorders, particularly mood disorders, other impulse-control disorders, and substance abuse or dependence (mainly nicotine dependence). CONCLUSIONS: Kleptomania presented a specific psychopathological profile that distinguished patients with this disorder from patients with alcohol abuse or dependence and other psychiatric comparison patients. Impulsivity was the major psychopathological feature of kleptomania. A link between kleptomania and affective disorder was supported by the high rate of comorbid affective disorders in patients with kleptomania and a specific pattern of variation in the two conditions over time. Further prospective studies are needed to confirm this pattern. Because kleptomania is characterized by a low rate of comorbid substance related disorders other than nicotine dependence and by severe psychopathology, it could be an appropriate disorder in which to study the information processes and psychobiology underlying impulsivity. PMID- 12900317 TI - Untreated depression and hippocampal volume loss. AB - OBJECTIVE: The purpose of this study was to investigate the effect of antidepressant treatment on hippocampal volumes in patients with major depression. METHOD: For 38 female outpatients, the total time each had been in a depressive episode was divided into days during which the patient was receiving antidepressant medication and days during which no antidepressant treatment was received. Hippocampal gray matter volumes were determined by high resolution magnetic resonance imaging and unbiased stereological measurement. RESULTS: Longer durations during which depressive episodes went untreated with antidepressant medication were associated with reductions in hippocampal volume. There was no significant relationship between hippocampal volume loss and time depressed while taking antidepressant medication or with lifetime exposure to antidepressants. CONCLUSIONS: Antidepressants may have a neuroprotective effect during depression. PMID- 12900316 TI - Clozapine as a first treatment for schizophrenia. AB - OBJECTIVE: The authors' goal was to explore whether clozapine given as the first antipsychotic treatment favorably affects the course of schizophrenia. METHOD: Thirty-four inpatients experiencing their first episode of schizophrenia or schizoaffective disorder were treated first with clozapine and then followed for up to 4 years. In a previous study, the authors followed patients experiencing their first episode of schizophrenia or schizoaffective disorder who were given fluphenazine as the first treatment. In the current study and the previous study, response criteria required sustained remission of positive symptoms. RESULTS: Nineteen of the 34 subjects met response criteria while taking clozapine. The median time to treatment response was 11 weeks (range=2-13). Using survival analysis, the authors determined that the cumulative response rate for the 34 patients was 66.4% at the end of 13 weeks, which is comparable to the response rate to fluphenazine in the previous study. All responses to clozapine occurred by 13 weeks. Eight (42%) of the clozapine responders discontinued clozapine before 6 months, and only six (32%) remained on clozapine for all of their time in the study. CONCLUSIONS: The authors found no benefit for clozapine over conventional antipsychotics for acute treatment of the first episode of schizophrenia or schizoaffective disorder. Long-term benefits could not be studied because of the high rate of early discontinuation of clozapine treatment. PMID- 12900318 TI - Short-term use of estradiol for depression in perimenopausal and postmenopausal women: a preliminary report. AB - OBJECTIVE: The authors examined the effect of a 4-week course of estrogen therapy on depression in perimenopausal and postmenopausal women. METHOD: Twenty-two depressed women who were either perimenopausal (N=10) or postmenopausal (N=12) received open-label treatment with transdermal 17beta-estradiol (100 micro g/day) for 4 weeks. The Montgomery-Asberg Depression Rating Scale and the Beck Depression Inventory were used to assess depressive symptoms, the Greene Climacteric Scale was used to assess menopause-related symptoms, and the Clinical Global Impression (CGI) was used to assess global clinical improvement in these women at baseline and after treatment. Remission of depression was defined as a score <10 on the Montgomery-Asberg Depression Rating Scale and a score 80% of patients undergoing catheter ablation. Moreover, the isolation of PVs is not crucial for curing AF. PMID- 12900337 TI - Intra-atrial pressure increases rate and organization of waves emanating from the superior pulmonary veins during atrial fibrillation. AB - BACKGROUND: Atrial fibrillation (AF) commonly associates with atrial dilatation by poorly understood mechanisms. We hypothesized that elevation of intra-atrial pressure elicits high-frequency and spatio-temporally organized left atrial (LA) sources emanating from the superior pulmonary veins. METHODS AND RESULTS: We used a stretch-related AF model in the sheep heart to induce stable episodes of AF (>40 minutes) in 9 animals. Video movies of the LA free wall (LAFW) and LA superior pulmonary vein junction (JPV) were obtained by using di-4-ANEPPS. Electrograms from the right atrium were recorded. At intra-atrial pressures >10 cm H2O, the maximum dominant frequency (DFMax) was significantly higher in the JPV than in the LAFW (12.0+/-0.2 and 10.5+/-0.2 Hz, respectively [mean+/-SEM]; P<0.001). Below 10 cm H2O, DFMax was similar in the JPV and LAFW (10.8+/-0.3 versus 10.2+/-0.3 Hz; P=0.6); DFMax in both JPV and LAFW was significantly higher than in the right atrium (7.8+/-0.3 Hz; P<0.001). Analysis of excitation direction in JPV showed positive correlation between intra-atrial pressure and the number of waves emanating from the left superior pulmonary vein (r=0.79, P=0.02) but not from the LAFW (r=0.54, P=0.09). The number of spatio-temporally periodic waves in the JPV correlated with pressure (r=0.92, P=0.002). In 3 cases, JPV rotors were identified with a cycle length equal to 1/DFMax. CONCLUSIONS: We demonstrate for the first time that an increase in intra-atrial pressure increases the rate and organization of waves emanating from the superior pulmonary veins underlying stretch-related AF. PMID- 12900338 TI - A20 is dynamically regulated in the heart and inhibits the hypertrophic response. AB - BACKGROUND: Nuclear factor (NF)-kappaB signaling has been implicated in cardiomyocyte hypertrophy. Here, we determine the cardiac regulation and biological activity of A20, an inhibitor of NF-kappaB signaling. METHODS AND RESULTS: Mice were subjected to aortic banding, and A20 expression was examined. A20 mRNA upregulation (4.3+/-1.5-fold; P<0.05) was detected 3 hours after banding, coinciding with peak NF-kappaB activation. A20 was also upregulated in cultured neonatal cardiomyocytes stimulated with phenylephrine or endothelin-1 (2.8+/-0.6- and 4+/-1.1-fold, respectively; P<0.05), again paralleling NF-kappaB activation. Infection of cardiomyocytes with an adenoviral vector (Ad) encoding A20 inhibited tumor necrosis factor-alpha-stimulated NF-kappaB signaling with an efficacy comparable to dominant negative inhibitor of kappa-B kinase beta (dnIKKbeta). Ad.dnIKKbeta-infected cardiomyocytes exhibited increased apoptosis when they were serum starved or subjected to hypoxia-reoxygenation, whereas Ad.A20-infected cardiomyocytes did not. Expression of Ad.A20 inhibited the hypertrophic response in cardiomyocytes stimulated with phenylephrine or endothelin-1. CONCLUSIONS: A20 is dynamically regulated during acute biomechanical stress in the heart and functions to attenuate cardiac hypertrophy through the inhibition of NF-kappaB signaling without sensitizing cardiomyocytes to apoptotic cell death. PMID- 12900339 TI - Randomized study to assess the effectiveness of slow- and moderate-release polymer-based paclitaxel-eluting stents for coronary artery lesions. AB - BACKGROUND: Early clinical studies demonstrated the feasibility of local paclitaxel delivery in reducing restenosis after treatment of de novo coronary lesions in small patient populations. METHODS AND RESULTS: We conducted a randomized, double-blind trial of 536 patients at 38 medical centers evaluating slow-release (SR) and moderate-release (MR) formulations of a polymer-based paclitaxel-eluting stent (TAXUS) for revascularization of single, primary lesions in native coronary arteries. Cohort I compared TAXUS-SR with control stents, and Cohort II compared TAXUS-MR with a second control group. The primary end point was 6-month percent in-stent net volume obstruction measured by intravascular ultrasound. Secondary end points were 6-month angiographic restenosis and 6- and 12-month incidence of major adverse cardiac events, a composite of cardiac death, myocardial infarction, and repeat revascularization. At 6 months, percent net volume obstruction within the stent was significantly lower for TAXUS stents (7.9% SR and 7.8% MR) than for respective controls (23.2% and 20.5%; P<0.0001 for both). This corresponded with a reduction in angiographic restenosis from 17.9% to 2.3% in the SR cohort (P<0.0001) and from 20.2% to 4.7% in the MR cohort (P=0.0002). The incidence of major adverse cardiac events at 12 months was significantly lower (P=0.0192) in the TAXUS-SR (10.9%) and TAXUS-MR (9.9%) groups than in controls (22.0% and 21.4%, respectively), predominantly because of a significant reduction in repeat revascularization of the target lesion in TAXUS treated patients. CONCLUSIONS: Compared with a bare metal stent, paclitaxel eluting stents reduced in-stent neointimal formation and restenosis and improved 12-month clinical outcome of patients with single de novo coronary lesions. PMID- 12900341 TI - Effect of lower dose of oral conjugated equine estrogen on size and oxidative susceptibility of low-density lipoprotein particles in postmenopausal women. AB - BACKGROUND: Estrogen replacement therapy (ERT) has an antioxidant effect that opposes the oxidation of LDL. Oral ERT-induced increases in plasma triglyceride, however, are associated with decreased LDL size, which may counteract this antioxidant effect. Because lower doses of oral estrogen do not affect plasma triglyceride concentrations, LDL size might not change, and the antioxidant effect of estrogen might be preserved. We investigated whether a lower dose of oral estrogen could eliminate the adverse effects of high-dose oral ERT on the size and oxidative susceptibility of LDL in postmenopausal women. METHODS AND RESULTS: Postmenopausal women received no treatment or were treated with oral conjugated equine estrogen (CEE) 0.625 or 0.3125 mg/d for 3 months. CEE at a dose of 0.625 mg/d significantly increased plasma triglyceride concentrations and decreased LDL diameter, but the concentrations of LDL-derived thiobarbituric acid reactive substances (TBARS) and lag time for conjugated diene formation did not change. In contrast, 0.3125 mg of CEE did not affect plasma triglyceride concentrations or LDL diameter and significantly decreased LDL-derived TBARS concentrations and significantly prolonged LDL lag time. Estrogen-induced changes in LDL diameter correlated negatively with changes in plasma triglyceride (r= 0.44, P<0.01) and LDL-derived TBARS (r=-0.57, P<0.001) but positively with changes in LDL lag time (r=0.42, P<0.01). CONCLUSIONS: Because oral CEE at a dose of 0.3125 mg/d does not elevate plasma triglyceride, resulting in unchanged size of LDL particles that are resistant to oxidation, the antioxidant effect of estrogen can be preserved. PMID- 12900340 TI - Systemic delivery of bone marrow-derived mesenchymal stem cells to the infarcted myocardium: feasibility, cell migration, and body distribution. AB - BACKGROUND: Systemic delivery of bone marrow-derived mesenchymal stem cells (BM MSCs) is an attractive approach for myocardial repair. We aimed to test this strategy in a rat model after myocardial infarction (MI). METHODS AND RESULTS: BM MSCs were obtained from rat bone marrow, expanded in vitro to a purity of >50%, and labeled with 99mTc exametazime, fluorescent dye, LacZ marker gene, or bromodeoxyuridine. Rats were subjected to MI by transient coronary artery occlusion or to sham MI. 99mTc-labeled cells (4x10(6)) were transfused into the left ventricular cavity of MI rats either at 2 or 10 to 14 days after MI and were compared with sham-MI rats or MI rats treated with intravenous infusion. Gamma camera imaging and isolated organ counting 4 hours after intravenous infusion revealed uptake of the 99mTc-labeled cells mainly in the lungs, with significantly smaller amounts in the liver, heart, and spleen. Delivery by left ventricular cavity infusion resulted in drastically lower lung uptake, better uptake in the heart, and specifically higher uptake in infarcted compared with sham-MI hearts. Histological examination at 1 week after infusion identified labeled cells either in the infarcted or border zone but not in remote viable myocardium or sham-MI hearts. Labeled cells were also identified in the lung, liver, spleen, and bone marrow. CONCLUSIONS: Systemic intravenous delivery of BM MSCs to rats after MI, although feasible, is limited by entrapment of the donor cells in the lungs. Direct left ventricular cavity infusion enhances migration and colonization of the cells preferentially to the ischemic myocardium. PMID- 12900342 TI - Mechanism of complement activation and its role in the inflammatory response after thoracoabdominal aortic aneurysm repair. AB - BACKGROUND: Complement activation contributes to ischemia-reperfusion injury. Patients undergoing thoracoabdominal aortic aneurysm (TAAA) repair suffer extensive ischemia-reperfusion and considerable systemic inflammation. METHODS AND RESULTS: The degree and mechanism of complement activation and its role in inflammation were investigated in 19 patients undergoing TAAA repair. Patients undergoing open infrarenal aortic surgery (n=5) or endovascular descending aortic aneurysm repair (n=6) served as control subjects. Substantial complement activation was seen in TAAA patients but not in controls. C1rs-C1-inhibitor complexes increased moderately, whereas C4bc, C3bBbP, C3bc, and the terminal SC5b 9 complex (TCC) increased markedly after reperfusion, reaching a maximum 8 hours after reperfusion. Interleukin (IL)-1beta, tumor necrosis factor alpha (TNF alpha), and IL-8 increased significantly in TAAA patients but not in controls, peaking at 24 hours postoperatively and correlating closely with the degree of complement activation. IL-6 and IL-10 increased to a maximum 8 hours after reperfusion in the TAAA patients, were not correlated with complement activation, and increased moderately in the control subjects. Myeloperoxidase and lactoferrin increased markedly before reperfusion in all groups, whereas sICAM-1, sP selectin, and sE-selectin were unchanged. No increase was observed in complement activation products, IL-1beta, TNF-alpha, or IL-8 in a mannose-binding lectin (MBL)-deficient TAAA patient, whereas IL-6, IL-10, myeloperoxidase, and lactoferrin increased as in the controls. Two other MBL-deficient TAAA patients receiving plasma attained significant MBL levels and showed complement and cytokine patterns identical to the MBL-sufficient TAAA patients. CONCLUSIONS: The data suggest that complement activation during TAAA repair is MBL mediated, amplified through the alternative pathway, and responsible in part for the inflammatory response. PMID- 12900343 TI - Quality of life of children after repair of transposition of the great arteries. AB - BACKGROUND: We sought to assess quality of life of children with transposition of the great arteries (TGA) enrolled during transition in management strategy from atrial to arterial switch operation. METHODS AND RESULTS: Neonates enrolled by the Congenital Heart Surgeons Society in a prospective study of TGA between 1985 and 1989 were eligible. A Child Health Questionnaire was sent for completion by the child between February and June 2000. Data were compared with published normative values. Child Health Questionnaires were completed by 306 of 708 survivors at a mean age of 13+/-1 years. Diagnosis included TGA (n=202, 66%), TGA/ventricular septal defect (VSD) (n=84, 27%), and TGA/VSD/pulmonary stenosis (n=20, 7%). Repair type was arterial switch (n=189, 62%), atrial switch (n=105, 34%; Senning=58, Mustard=47), or Rastelli (n=12, 4%). Children with TGA scored significantly higher than published norms in all categories except self-esteem. TGA/VSD/pulmonary stenosis was associated with lower scores than TGA and TGA/VSD in physical functioning (P=0.002), general health perceptions (P=0.012), and mental health (P=0.048). Arterial repair was associated with higher scores than atrial or Rastelli repair in physical functioning (P<0.001), pain (P=0.004), mental health (P=0.019), self-esteem (P=0.004), and general health perceptions (P<0.001). By multivariable analyses, the most common independent factors impacting scores were repair type, perfusion parameters, and gender. CONCLUSIONS: Quality of life and health status as perceived by children 11 to 15 years after TGA repair is excellent when compared with published normative data and is better after arterial switch operation than after atrial repair. PMID- 12900344 TI - Plasma carotenoids and tocopherols and risk of myocardial infarction in a low risk population of US male physicians. AB - BACKGROUND: Increased intake of carotenoids and vitamin E may protect against myocardial infarction (MI). However, prospective data on blood levels of carotenoids other than beta-carotene and vitamin E (tocopherol) and risk of MI are sparse. METHODS AND RESULTS: We conducted a prospective, nested case-control analysis among male physicians without prior history of cardiovascular disease who were followed for up to 13 years in the Physicians' Health Study. Samples from 531 physicians diagnosed with MI were analyzed together with samples from paired control subjects, matched for age and smoking, for 5 major carotenoids (alpha- and beta-carotene, beta-cryptoxanthin, lutein, and lycopene), retinol, and alpha- and gamma-tocopherol. Overall, we found no evidence for a protective effect against MI for higher baseline plasma levels of retinol or any of the carotenoids measured. Among current and former smokers but not among never smokers, higher baseline plasma levels of beta-carotene tended to be associated with lower risk (P for interaction=0.02). Men with higher plasma levels of gamma tocopherol tended to have an increased risk of MI (P for trend=0.01). CONCLUSIONS: These prospective data do not support an overall protective relation between plasma carotenoids or tocopherols and future MI risk among men without a history of prior cardiovascular disease. PMID- 12900345 TI - Cost effectiveness of defibrillation by targeted responders in public settings. AB - BACKGROUND: Out-of-hospital cardiac arrest is frequent and has poor outcomes. Defibrillation by trained targeted nontraditional responders improves survival versus historical controls, but it is unclear whether such defibrillation is a good value for the money. Therefore, this study estimated the incremental cost effectiveness of defibrillation by targeted nontraditional responders in public settings by using decision analysis. METHODS AND RESULTS: A Markov model evaluated the potential cost effectiveness of standard emergency medical services (EMS) versus targeted nontraditional responders. Standard EMS included first responder defibrillation followed by advanced life support. Targeted nontraditional responders included standard EMS supplemented by defibrillation by trained lay responders. The analysis adopted a US societal perspective. Input data were derived from published or publicly available data. Future costs and effects were discounted at 3%. Monte Carlo simulation and sensitivity analyses assessed the robustness of results. Standard EMS had a median of 0.47 (interquartile range [IQR]=0.32 to 0.69) quality-adjusted life years and a median of 14 100 dollars (IQR=8600 dollars to 21 900 dollars) costs per arrest. Targeted nontraditional responders in casinos had an incremental cost of a median 56 700 dollars (IQR=44 100 dollars to 77 200 dollars) per additional quality-adjusted life year. The results were sensitive to changes in time to defibrillation, incidence of arrest, and number of devices required to implement rapid defibrillation. CONCLUSIONS: Where cardiac arrest is frequent and response time intervals are short, rapid defibrillation by targeted nontraditional responders may be a good value for the money compared with standard EMS. The incidence of arrest should be considered when choosing locations to implement public access defibrillation. PMID- 12900346 TI - Detectable serum cardiac troponin T as a marker of poor prognosis among patients with chronic precapillary pulmonary hypertension. AB - BACKGROUND: Right ventricular failure is a leading cause of death in patients with chronic pulmonary hypertension (PH). We checked whether detection of cardiac troponin T (cTnT), a specific marker of myocyte injury, could be useful in prognostic stratification of those patients. METHODS AND RESULTS: Initial evaluation of 56 clinically stable patients (age 41+/-15 years) with pulmonary arterial (51 patients) or inoperable chronic thromboembolic (5 patients) PH (mean pulmonary arterial pressure 60+/-18 mm Hg) included cTnT test, allowing detection of its serum levels > or =0.01 ng/mL [cTnT(+)]. cTnT was detectable in 8 of 56 (14%) patients (mean+/-SD, 0.034+/-0.022; range, 0.010 to 0.077 ng/mL). Despite similar pulmonary hemodynamics, they had higher heart rate (92+/-15 versus 76+/ 14 bpm, P=0.004), lower mixed venous oxygen saturation (50+/-10% versus 57+/-9%, P=0.04), and higher serum N-terminal pro-B-type natriuretic peptide (4528+/-3170 versus 2054+/-2168 pg/mL, P=0.03) and walked less during the 6-minute walk test (298+/-132 versus 396+/-101 m, P=0.02). Cumulative survival estimated by Kaplan Meier curves was significantly worse at 24 months in cTnT(+) compared with cTnT( ) (29% versus 81%, respectively, log-rank test P=0.001). Multivariate analysis revealed cTnT status (hazard ratio, 4.89; 95% CI, 1.18 to 20.29; P=0.03), 6 minute walk test (hazard ratio, 0.93 for each 10 m; P=0.01), and pulmonary vascular resistance (hazard ratio, 1.13; P=0.01) as independent markers of mortality. All 3 cTnT(+) patients who survived the follow-up period converted to cTnT(-) during treatment. CONCLUSIONS: Detectable cTnT is a so-far ignored independent marker of increased mortality risk in patients with chronic precapillary PH, supporting the role of progressive myocyte injury in the vicious circle leading to hemodynamic destabilization. PMID- 12900347 TI - Effects of unilateral and bilateral carotid baroreflex stimulation on cardiac and neural sympathetic discharge oscillatory patterns. AB - BACKGROUND: Left and right carotid baroreflex afferents participate in generating the spontaneous variability of heart rate (HR), arterial pressure (AP), and muscle sympathetic nerve activity (MSNA), but the relative contribution of each side is unclear. Pathophysiological conditions unilaterally affecting carotid baroreceptor function might result in abnormal changes of HR, AP, and MSNA variability, thus markedly affecting prognosis. We tested the hypothesis that unilateral carotid baroreceptor perturbation might differentially affect HR, AP, and MSNA variability compared with stimulation of the opposite side. METHODS AND RESULTS: In 12 healthy volunteers, 4 sinusoidal neck suction procedures (0.1 Hz, from 0 to -50 mm Hg) were applied at the right, left, and combined right and left sides of the neck, in concordance or with phase opposition. Respiration was controlled at 0.25 Hz. Power spectrum analysis assessed the changes in the 0.1-Hz oscillatory component of the R-R interval, systolic AP (SAP), and MSNA variability induced by rhythmic baroreceptor stimulation. Mean R-R interval, SAP, and MSNA were unchanged during all procedures. The increase of the 0.1-Hz component of R-R and SAP variability during right and combined right and left carotid baroreceptor stimulation was greater than the changes induced by left sided stimulation. The increase in the 0.1-Hz oscillatory component of MSNA variability was similar during all neck suction procedures. CONCLUSIONS: Right carotid baroreflex loading was as efficient as bilateral stimulation and more effective than left carotid suction in modulating R-R and SAP variability. There was no asymmetry in neural sympathetic discharge responses after single-sided carotid baroreceptor stimulation. PMID- 12900348 TI - Role of cyclooxygenase-2 in the generation of vasoactive prostanoids in the rat pulmonary and systemic vascular beds. AB - BACKGROUND: Prostanoid synthesis by the cyclooxygenase (COX)-2 pathway plays an important role in inflammation, and recent studies have shown the presence of COX 2 in the normal rat lung. However, the role of COX-2 in the generation of vasoactive prostanoids in the rat is uncertain. In the present study, the hypothesis that synthesis of vasoactive prostanoids via the COX-2 pathway can alter pulmonary and systemic vascular resistance was investigated, and the effects of selective COX-2 inhibitors on pulmonary and systemic responses to the prostanoid precursor arachidonic acid were examined in the anesthetized rat with a recently developed right-heart catheterization technique. METHODS AND RESULTS: Injections of arachidonic acid caused dose-related increases in pulmonary vascular resistance and decreases in systemic vascular resistance. These responses were attenuated by selective COX-2 inhibitors and a selective COX-1 inhibitor, whereas responses to exogenous prostanoids were not altered. Nimesulide or NS-398 did not alter arachidonic acid-induced platelet aggregation in rat platelet-rich plasma. Western blot analysis and immunostaining showed the expression of both COX isoforms in the rat lung. CONCLUSIONS: The results of these experiments suggest that arachidonic acid is converted into vasoactive prostanoids by the COX-2 and COX-1 pathway in the pulmonary and peripheral vascular beds in the rat and that TXA2 is a major prostanoid formed in the normal rat lung. PMID- 12900349 TI - Human cytomegalovirus seropositivity is associated with impaired vascular function. AB - BACKGROUND: Herpesvirus infection is a possible risk factor for atherogenesis, and diabetics may be at particular risk. Endothelial dysfunction is an early marker for atherosclerosis, and the present study tests the hypotheses that (1) prior infection with cytomegalovirus (CMV) and herpes simplex virus (HSV) is associated with endothelial dysfunction and (2) this may be more marked in diabetics. METHODS AND RESULTS: Serum samples were tested for anti-IgG antibodies to CMV and HSV from 400 subjects (mean age for diabetics and nondiabetics, 37.8+/ 4.3 and 37.9+/-3.7 [SD]). We also assessed Helicobacter pylori and Chlamydia pneumoniae serology. Coronary atheroma was quantified by means of electron beam computed tomography. Subjects (n=157) underwent venous occlusion plethysmography with acetylcholine, bradykinin, glyceryl trinitrate, norepinephrine, and l-NG monomethyl-l-arginine. Individuals who were seropositive for CMV had reduced responses to bradykinin (P=0.005) and glyceryl trinitrate (P=0.006). The reduced response to bradykinin remained significant (P=0.045) after adjusting for the response to glyceryl trinitrate and was independent of conventional risk factors. Positive serology for the other organisms did not have an independent effect on reactivity. There was a weaker association between CMV and coronary artery calcification (P=0.09). Positive serology for each of the other pathogens did not affect reactivity, but there was a relation between total pathogen burden and impaired vascular reactivity. No significant differences were found between diabetics and nondiabetics. CONCLUSIONS: This study shows that CMV-seropositive individuals have endothelial dysfunction and impaired responses to NO. This association was independent of conventional risk factors and may be associated with increased atherosclerosis burden. PMID- 12900350 TI - Amniotic fluid as a novel source of mesenchymal stem cells for therapeutic transplantation. PMID- 12900351 TI - In B-cell chronic lymphocytic leukemias, 7q21 translocations lead to overexpression of the CDK6 gene. PMID- 12900352 TI - Evaluating the relevance of the platelet transcriptome. PMID- 12900353 TI - Does cytogenetic mosaicism in CD34+CD38low cells reflect the persistence of normal primitive hematopoietic progenitors in myeloid metaplasia with myelofibrosis? PMID- 12900354 TI - Aberrant BCL10 nuclear expression in nasal NK/T-cell lymphoma. PMID- 12900355 TI - Silencing of the SOCS-1 gene by CpG methylation? PMID- 12900356 TI - Acute chest syndrome, transfusion, and neurologic events in children with sickle cell disease. PMID- 12900357 TI - Rituximab-associated immune myelopathy. PMID- 12900358 TI - Vagaries of genetic association studies in myocardial infarction. PMID- 12900359 TI - Pathologic response to preoperative systemic therapy: the new biological paradigm for predicting outcome and planning therapy. PMID- 12900360 TI - Preoperative chemotherapy for breast cancer: do we need to redefine our end points? PMID- 12900361 TI - Perspectives and risks of breast-conservation therapy for breast cancer. PMID- 12900362 TI - Angiogenesis and antiangiogenic therapy of colon cancer liver metastasis. AB - The fact that tumor growth and metastatic spread relies on angiogenesis has been widely proven and accepted. The understanding of cancer biology and metastasis formation has led to the development of new therapeutic approaches that target tumor biology. The survival and establishment of metastatic lesions depend on a shift in the normal balance of proangiogenic and antiangiogenic factors that favor angiogenesis. Colorectal cancer is one of the leading cancer deaths worldwide. Angiogenesis has been associated with colon cancer progression and metastatic spread, thereby significantly affecting patient survival. New experimental approaches that inhibit angiogenic processes have demonstrated promising antineoplastic effects on metastatic colorectal cancer and are partially being investigated in clinical trials. This review focuses on angiogenesis in colorectal cancer metastasis formation as a target for antiangiogenic therapy, describing the experience from experimental studies and current clinical trials. PMID- 12900363 TI - Histopathologic evidence of tumor regression in the axillary lymph nodes of patients treated with preoperative chemotherapy correlates with breast cancer outcome. AB - BACKGROUND: The benefits of primary tumor downstaging and assessment of chemoresponsiveness have resulted in expanded applications for induction chemotherapy. However, the pathologic evaluation and prognostic significance of response in preoperatively treated lymph nodes have not been defined. METHODS: The axillary lymph nodes of 71 patients with locally advanced breast cancer treated with induction chemotherapy were evaluated for histological evidence of tumor regression as defined by the presence of nodal fibrosis, mucin pools, or aggregates of foamy histiocytes. RESULTS: Complete pathologic response in the breast and axilla occurred in 10 patients (14%); 19 (26.8%) had evidence of tumor regression in 1 or more lymph nodes. Patients without nodal metastases and no evidence of tumor regression had the best outcome (median disease-free survival, 31.5 months; relapse rate, 27%). Patients with residual nodal metastases and no evidence of treatment effect had the worst outcome (median disease-free survival, 19.8 months; relapse rate, 55%). The median disease-free survival was 22.1 months, and the relapse rate was 32% for patients with histopathologic evidence of tumor regression in the axillary lymph nodes. CONCLUSIONS: Detection of treatment effect in axillary lymph nodes after induction chemotherapy identifies a subset of patients with an outcome intermediate between that of completely node negative and node-positive patients. The axillary lymph nodes of patients receiving preoperative chemotherapy should be routinely analyzed for the presence of these features. PMID- 12900364 TI - Small invasive breast carcinomas in Taiwanese women. AB - BACKGROUND: Female Taiwanese breast cancer patients are younger than their Western counterparts. This study examined the predictors of axillary lymph node metastases in Taiwanese women with T1 breast cancer. METHODS: Data from 394 Taiwanese women with T1 invasive breast carcinoma were retrospectively reviewed. RESULTS: The data contained 6 T1a, 51 T1b, and 337 T1c breast tumors. The patients' ages ranged from 23 to 82 years (mean +/- SD, 48.2 +/- 11.4 years; median, 46.4 years). Axillary nodal metastases were present in 38.3% of the patients (16.7% in T1a, 35.3% in T1b, and 39.2% in T1c tumors). The patients with nodal metastases had significantly greater body weights and S-phase fractions than those without nodal metastases. Univariate analysis revealed that unfavorable pathology, lymphovascular invasion, S-phase fraction >7%, and nondiploid DNA ploidy were significantly associated with lymph node metastases. Lymphovascular invasion was the only significant variable as the independent predictor in the multiple logistic regression analysis. In the Cox proportional hazards regression analysis, axillary nodal status and lymphovascular invasion were significantly associated with survival. CONCLUSIONS: Taiwanese women with small breast cancer displayed a relatively higher incidence of axillary lymph node metastases than Western women. Axillary lymph node dissection or sentinel lymph node biopsy should be conducted on Taiwanese patients with small invasive breast carcinomas, particularly when risk factors exist. PMID- 12900365 TI - The finding of invasive cancer after a preoperative diagnosis of ductal carcinoma in-situ: causes of ductal carcinoma-in-situ underestimates with stereotactic 14 gauge needle biopsy. AB - BACKGROUND: For the evaluation of nonpalpable lesions of the breast, image-guided 14-gauge automated needle biopsy is increasingly replacing wire-localized excision. When ductal carcinoma-in-situ (DCIS) is diagnosed at core biopsy, invasive cancer is found in approximately 17% of excision specimens. These so called DCIS underestimates pose a problem for patients and surgeons, because they generally cause extension of treatment. We evaluated DCIS underestimates in detail to assess reasons for missing the invasive component at core biopsy. This evaluation also included a histological comparison with true DCIS (DCIS at core biopsy and excision). METHODS: Between 1997 and 2000, DCIS was diagnosed at 14 gauge needle biopsy in 255 patients. In 41 cases (16%), invasive cancer was found at excision. We performed a thorough histopathological and radiological review of all these DCIS underestimates, including a histological comparison with core biopsy specimens of 32 true DCIS cases. We assessed the main reason for missing the invasive component at core biopsy. RESULTS: Causes for DCIS underestimates were categorized into "mainly radiological" (n = 20), "mainly histopathological" (n = 15), and "histological disagreements" (n = 6). High-grade DCIS and periductal inflammation in core biopsies made a DCIS underestimate 2.9 and 3.3 times more likely, respectively. CONCLUSIONS: A variety of radiological and histopathological reasons contribute to the DCIS underestimate rate. Approximately half of these are potentially avoidable. PMID- 12900366 TI - Neoadjuvant treatment for resectable cancer of the esophagus and the gastroesophageal junction: a meta-analysis of randomized clinical trials. AB - BACKGROUND: There is no general agreement on the effect of neoadjuvant treatment for esophageal cancer on patient survival. METHODS: A meta-analysis was performed to determine the effect of preoperative treatment on survival of patients with resectable esophageal cancer and the effect of preoperative treatment on patient mortality. A standard variance-based method was used to derive summary estimates of the absolute difference in both 2-year survival and treatment-related mortality. RESULTS: Eleven randomized trials involving 2311 patients were analyzed. Preoperative chemotherapy improved 2-year survival compared with surgery alone: the absolute difference was 4.4% (95% confidence interval [CI],.3% 8.5%). Marginal evidence of heterogeneity was eliminated by restricting attention to the four most recent studies, which increased the estimate to 6.3% (95% CI, 1.8%-10.7%). For combined chemoradiotherapy, the increase was 6.4% (nonsignificant; 95% CI, -1.2%-14.0%). Treatment-related mortality increased by 1.7% with neoadjuvant chemotherapy (95% CI, -.9%-4.3%) and by 3.4% with chemoradiotherapy (95% CI, -.1%-7.3%), compared with surgery alone. CONCLUSIONS: There seems to be a modest survival advantage for patients who receive neoadjuvant chemotherapy followed by surgery, as compared with surgery alone. There is an apparent increase in treatment-related mortality, mainly for patients who receive neoadjuvant chemoradiotherapy. PMID- 12900368 TI - Radiofrequency tissue ablation: effect of hepatic blood flow occlusion on thermal injuries produced in cirrhotic livers. AB - BACKGROUND: Radiofrequency thermal ablation has been used as a treatment for several types of hepatic malignancies. Many of these lesions exist in the presence of cirrhosis. Limitations exist to the size of the ablations and, subsequently, the efficacy of treatment. Hepatic vascular inflow occlusion has been advocated as an adjunctive measure to increase the efficacy of the ablation. We present a model in the human cirrhotic liver that demonstrates the advantage of blood flow occlusion during radiofrequency ablation. METHODS: Five patients with advanced endstage liver disease scheduled to have orthotopic liver transplantation were enrolled in this study. After laparotomy and before hepatectomy, radiofrequency ablation was performed without and with hepatic blood flow occlusion. After hepatectomy, the liver was sectioned, the area of ablation was measured in three dimensions, and the volume of ablation calculated. RESULTS: Three of the patients had had previously placed transjugular intrahepatic portosystemic shunt. The mean volume of the ablation without blood flow occlusion was 22.5 +/- 7.4 cm(3) and that with blood flow occlusion was 48.4 +/- 24.0 cm(3) (P =.05). CONCLUSIONS: Ablation area is increased significantly with hepatic blood flow occlusion in the human cirrhotic liver. This result may have application in the treatment of larger (>3 cm) hepatic malignancies. PMID- 12900367 TI - Gene therapy of pancreatic cancer with green fluorescent protein and tumor necrosis factor-related apoptosis-inducing ligand fusion gene expression driven by a human telomerase reverse transcriptase promoter. AB - BACKGROUND: Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) induces apoptosis in malignant cells but not in normal cells. Ad/g-TRAIL, an adenoviral vector in which expression of green fluorescent protein (GFP) and TRAIL is driven by a human telomerase reverse transcriptase promoter, has shown promise as a targeted antitumor agent. METHODS: To investigate the effects of TRAIL gene therapy on pancreatic cancer, BxPC-3, MIA-PaCa-2, Panc-1, and ASPC-1 cells were treated with Ad/g-TRAIL. Transfection and protein expression were determined by using immunoblotting and identification of GFP with fluorescent microscopy and flow cytometry. Cell viability was determined by proliferation assay. Cell-cycle analysis and quantification of caspase-3 were used to identify apoptosis. The in vivo efficacy of Ad/g-TRAIL was characterized in a novel red fluorescent protein murine model of MIA-PaCa-2 pancreatic cancer. RESULTS: Cells treated with Ad/g-TRAIL expressed GFP and exhibited apoptotic morphology within 2 days of treatment. Treatment with this vector in vitro resulted in less cell viability, increased caspase-3 activity, and a greater apoptotic fraction than treatment with controls. In vivo, treatment with Ad/g-TRAIL significantly suppressed tumor growth. CONCLUSIONS: TRAIL gene therapy induces apoptosis of pancreatic tumor cells both in vitro and in vivo and is a promising therapy in the treatment of pancreatic cancer. PMID- 12900369 TI - Prospective identification of risk factors for wound infection after lower extremity oncologic surgery. AB - BACKGROUND: Surgical site infections (SSI) are frequent causes of morbidity and mortality after orthopaedic oncologic procedures. This study was conducted to identify the surgical site infection rate following a lower extremity or pelvic procedure and assess the risk factors for acquiring SSI by direct observation of orthopaedic oncology patients' wounds at a comprehensive cancer center. METHODS: One hundred ten consecutive patients were prospectively studied. The surveillance of surgical site infections was carried out by a surgeon-trained nurse from the Infectious Disease Service. Nineteen variables were analyzed as risk factors. RESULTS: The overall SSI rate was 13.6% (15 of 110). Excluding those patients with known preoperative infections, the SSI rate was 9.5% (10 of 105). Two statistically significant risk factors for surgical site infection in these patients emerged in the multivariate analysis: blood transfusion (P =.007) and obesity (P =.016). Procedure category was significant in univariate analysis only. Preoperative length of stay, length of procedure, prior adjuvant treatment (chemotherapy or radiotherapy), prior surgery, and use of an implant or allograft were not statistically significant risk factors for wound infection. Antibiotic usage patterns did not influence SSI rate. CONCLUSIONS: Blood transfusion and obesity should be considered individual risk factors for the development of wound infection in patients having orthopaedic oncologic procedures. PMID- 12900370 TI - Accurate lymph node staging is of greater prognostic importance than subclassification of the T2 category for gastric adenocarcinoma. AB - BACKGROUND: Examination of 15 or more lymph nodes is necessary for accurate staging of gastric adenocarcinoma. This study examined whether prognostic discrimination is improved by distinguishing between pT2 tumors limited to the muscularis propria (MP) and those extending to subserosa (SS). METHODS: A single institution, prospectively maintained database for 1985-2000 was reviewed for patients who had had R0 resection of pT2 gastric carcinoma. RESULTS: There were 161 patients with MP and 201 patients with SS. The prevalence of nodal metastasis was significantly lower with MP than with SS (47% vs. 66%, respectively; P <.001). As compared with MP, SS was associated with a similar prevalence of pN1 (44% vs. 43%) but a significantly higher prevalence of pN2 or pN3 (3% vs. 23%; P <.001). Five-year survival was significantly greater for patients with MP than with SS (64% vs. 49%; P =.005). On multivariate analyses, however, only the pN category and tumor site were independently significant prognostic factors. With accurate nodal staging, patients with MP or SS had similar survival in association with pN0 (90% vs. 86%) or pN1 (56% vs. 44%). pN2 or pN3 were mainly limited to SS tumors and these patients had poor survival (26% and 0%). CONCLUSION: For pT2 gastric adenocarcinoma, the depth of mural invasion was not an independently significant prognostic factor. PMID- 12900371 TI - Cell-cycle regulators and the Ki-67 labeling index can predict the response to chemoradiotherapy and the survival of patients with locally advanced squamous cell carcinoma of the esophagus. AB - BACKGROUND: We investigated whether aberrant p53 and p16 expression, the Ki-67 labeling index (LI), and int-2/cyclin D1 gene amplification predict the response to chemoradiotherapy (CRT) in patients with locally advanced esophageal squamous cell carcinoma (ESCC). METHODS: p53 and p16 expression status, the Ki-67 LI, and int-2/cyclin D1 amplification were assessed by immunohistochemical staining and slot blot analysis in pretreatment endoscopic biopsy specimens of 41 patients with T4 or M1 Lym (distant lymph node metastasis) ESCC. All patients received a course of chemotherapy (5-fluorouracil and cisplatin) with radiotherapy. RESULTS: The CRT therapeutic response rate was 71%, and resection after CRT was successful in 15 of the cases in which the CRT effect was significant. The cumulative survival rate after CRT in the p53-negative patients was significantly higher than in the p53-positive patients (P =.037). The mean Ki-67 LI in the CRT response cases was significantly higher than in the CRT no-response cases (P =.023). Multivariate regression analysis revealed high Ki-67 LI to be an independent variable linked to a pathologic complete response to CRT (P =.033). The cumulative survival rate after CRT in the group that was p53-negative and int 2/cyclin D1 amplification-positive was significantly higher than in the other groups (P =.008). CONCLUSIONS: Evaluating predictive factors in pretreatment endoscopic biopsy specimens may allow selection of more suitable multimodal treatment for ESCC patients and improve their quality of life. PMID- 12900372 TI - Decreased expression of stem cell factor in esophageal and gastric mucosa after esophagogastric anastomosis for cancer: potential relevance to motility. AB - BACKGROUND: Esophageal replacement with gastric tube is a well-established reconstruction of the alimentary tract after esophagectomy in cancer patients. The resulting molecular events in the transposed gastric tube and residual esophagus have yet to be investigated. Stem cell factor (SCF) was recently shown to be critical for signaling in gastrointestinal motility. SCF expression is here correlated with changes in mucosal morphology, acid and biliary reflux, and motility in the residual esophagus and gastric tube. METHODS: Thirteen patients surgically resected for squamous esophageal carcinoma with gastric tube replaced by esophagogastric anastomosis underwent upper endoscopy, esophageal manometry, 24-hour pH monitoring, and bile reflux detection. Esophageal and gastric mucosa samples were examined for SCF expression by immunohistochemical and semiquantitative reverse transcriptase-polymerase chain reaction analysis and for SCF serum levels by enzyme-linked immunosorbent assay. RESULTS: All patients showed severe residual esophagus hypoperistalsis and no gastric tube motor activity. The 24-hour pH monitoring was positive in most; 24-hour bile detection was mostly negative. SCF levels in the residual esophageal and gastric tube mucosa were dramatically decreased compared with those of normal subjects. The correlation between SCF and slow-wave activity was positive. CONCLUSIONS: Hypomotility of the residual esophagus and gastric tube seems closely associated with disruption of the SCF/c-kit signaling pathway. However, the absence of notable relations between mucosal changes after chronic exposure to acid, biliary gastric content, and SCF expression indicates that this analysis cannot be considered part of endoscopic follow-up. PMID- 12900373 TI - Synergistic effect of a granulocyte-macrophage colony-stimulating factor transduced tumor vaccine and systemic interleukin-2 in the treatment of murine colorectal cancer hepatic metastases. AB - BACKGROUND: Granulocyte-macrophage colony-stimulating factor-transduced tumor cell vaccines are less effective against cancer as the interval between metastasis and the initial vaccination increases. METHODS: Hepatic metastases were generated in BALB/c mice by using a syngeneic colorectal cancer line (CT26) with a splenic injection model. Irradiated CT26 cells transduced to secrete granulocyte-macrophage colony-stimulating factor were used as vaccine. Treatment groups received vaccine, systemic interleukin (IL-2), or both. Livers were examined for gross metastases 21 days after tumor challenge. Splenocytes were analyzed for in vitro activity against CT26 by using an enzyme-linked immunospot assay and a cytotoxic T lymphocyte assay. RESULTS: Eighty-eight percent of mice treated with vaccines and IL-2 were tumor free on day 21 (P 500 mg/dl in STZ-injected mice. BTC (0.2 microg/g) significantly reduced the plasma glucose concentration, but a higher concentration was ineffective. The effect of BTC was marked by day 4 but became smaller on day 6 or later. The plasma insulin concentration and the insulin content were significantly higher in mice treated with 0.1 and 0.2 microg/g BTC. BTC treatment significantly increased the number of beta-cells in each islet as well as the number of insulin-positive islets. Within islets, the numbers of 5-bromo-2-deoxyuridine/somatostatin-positive cells and pancreatic duodenal homeobox-1/somatostatin-positive cells were significantly increased by BTC. These results indicate that BTC improved hyperglycemia induced by a high dose of STZ by promoting neoformation of beta-cells, mainly from somatostatin-positive islet cells. PMID- 12900380 TI - O-glycosylation of Sp1 and transcriptional regulation of the calmodulin gene by insulin and glucagon. AB - Both insulin and glucagon stimulate steady-state levels of Sp1 transcription factor, but only insulin stimulates transcription of the calmodulin (CaM) gene in liver. Because O-glycosylation of Sp1 by O-linked N-acetylglucosamine (O-GlcNAc) is thought to regulate its ability to activate transcription, we assayed the levels of Sp1 with anti-Sp1 and anti-O-GlcNAc antibodies in Western blots by use of extracts of H-411E liver cells treated with insulin (10,000 microU/ml) or glucagon (1.5 x 10(-5) M). We also assessed subcellular localization of the native and glycosylated Sp1 in H411E cells treated with either hormone in the presence of deoxynorleucine (DON, an indirect inhibitor of O-glycosylation) or streptozotocin (STZ, an indirect stimulator of O-glycosylation). Insulin stimulated both total and O-GlcNAc-modified Sp1 primarily in the nucleus and induced CaM gene transcription (P < 0.0001). In contrast, glucagon promoted accumulation of Sp1 in the cytoplasm but not the nucleus, without significantly stimulating (P = not significant) either its O-glycosylation or transcription of the CaM gene. DON inhibited O-glycosylation of Sp1 and its ability to migrate to the nucleus and transactivate CaM gene transcription. In contrast, cotreatment of cells with STZ and glucagon enhanced O-glycosylation of Sp1, promoting its migration to the nucleus and resulting in increased CaM gene transcription. Thus O-glycosylation of Sp1 by insulin, but not glucagon, apparently enhances its (Sp1) nuclear recruitment and results in activation of CaM gene transcription. PMID- 12900381 TI - Activation of alpha2-adrenergic receptors blunts epinephrine-induced lipolysis in subcutaneous adipose tissue during a hyperinsulinemic euglycemic clamp in men. AB - The aim of this study was to investigate whether hyperinsulinemia modifies adrenergic control of lipolysis, with particular attention paid to the involvement of antilipolytic alpha2-adrenergic receptors (AR). Eight healthy male subjects (age: 23.9 +/- 0.9 yr; body mass index: 23.8 +/- 1.9) were investigated during a 6-h euglycemichyperinsulinemic clamp and in control conditions. Before and during the clamp, the effect of graded perfusions of isoproterenol (0.1 and 1 microM) or epinephrine (1 and 10 microM) on the extracellular glycerol concentration in subcutaneous abdominal adipose tissue was evaluated by using the microdialysis method. Both isoproterenol and epinephrine induced a dose-dependent increase in extracellular glycerol concentration when infused for 60 min through the microdialysis probes before and during hours 3 and 6 of the clamp. The catecholamine-induced increase was significantly lower during the clamp than before it, with the inhibition being more pronounced in hour 6 of the clamp. Isoproterenol (1 microM)-induced lipolysis was reduced by 28 and 44% during hours 3 and 6 of the clamp, respectively, whereas the reduction of epinephrine (100 microM)-induced lipolysis was significantly greater (by 63 and 70%, P < 0.01 and P < 0.04, respectively) during the same time intervals. When epinephrine was infused in combination with 100 microM phentolamine (a nonselective alpha-AR antagonist), the inhibition of epinephrine (10 microM)-induced lipolysis was only of 19 and 40% during hours 3 and 6 of the clamp, respectively. The results demonstrate that, in situ, insulin counteracts the epinephrine-induced lipolysis in adipose tissue. The effect involves 1) reduction of lipolysis stimulation mediated by the beta-adrenergic pathway and 2) the antilipolytic component of epinephrine action mediated by alpha2-ARs. PMID- 12900382 TI - T3 increases lactate transport and the expression of MCT4, but not MCT1, in rat skeletal muscle. AB - Triiodothyronine (T3) regulates the expression of genes involved in muscle metabolism. Therefore, we examined the effects of a 7-day T3 treatment on the monocarboxylate transporters (MCT)1 and MCT4 in heart and in red (RG) and white gastrocnemius muscle (WG). We also examined rates of lactate transport into giant sarcolemmal vesicles and the plasmalemmal MCT1 and MCT4 in these vesicles. Ingestion of T3 markedly increased circulating serum T3 (P < 0.05) and reduced weight gain (P < 0.05). T3 upregulated MCT1 mRNA (RG +77, WG +49, heart +114%, P < 0.05) and MCT4 mRNA (RG +300, WG +40%). However, only MCT4 protein expression was increased (RG +43, WG +49%), not MCT1 protein expression. No changes in MCT1 protein were observed in any tissue. T3 treatment doubled the rate of lactate transport when vesicles were exposed to 1 mM lactate (P < 0.05). However, plasmalemmal MCT4 was only modestly increased (+13%, P < 0.05). We conclude that T3 1) regulates MCT4, but not MCT1, protein expression and 2) increases lactate transport rates. This latter effect is difficult to explain by the modest changes in plasmalemmal MCT4. We speculate that either the activity of sarcolemmal MCTs has been altered or else other MCTs in muscle may have been upregulated. PMID- 12900383 TI - In vivo stimulation of sympathetic nervous system modulates osteoblastic activity in mouse calvaria. AB - Previously, we demonstrated that epinephrine induced the expression of interleukin (IL)-6 mRNA via beta-adrenoceptors in cultured human osteoblastic cells. IL-6 is well known to modulate bone metabolism by regulating the development and function of osteoclasts and osteoblasts. Recently, restraint stress and intracerebroventricular injection of lipopolysaccharide (LPS) have been reported to induce the expression of IL-6 mRNA in peripheral organs in mice in which expression is mediated by the activation of the sympathetic nervous system. To prove the physiological role of sympathetic nerves in bone metabolism in vivo, we examined by RT-PCR analysis the effects of restraint stress and intracerebroventricular injection of LPS on IL-6 mRNA expression in mouse calvaria. The expression of IL-6 mRNA in mouse calvaria was stimulated by either restraint stress (30 min) or intracerebroventricular injection of LPS (50 ng/mouse, 60 min). The treatment of mice with the neurotoxin 6-hydroxydopamine (6 OHDA, 100 mg x kg-1 x day-1 ip for 3 days) inhibited LPS (icv)-induced expression of IL-6 mRNA in their calvaria. The expression of IL-6 mRNA induced by the restraint stress was not influenced by 6-OHDA, which destroys noradrenergic nerve terminals. Furthermore, pretreatment with a beta-blocker, propranolol (15 or 25 mg/kg ip), inhibited both stress- and LPS-induced increases in the level of IL-6 mRNA, but pretreatment with an alpha-blocker, phentolamine (5 mg/kg sc), did not inhibit them in mouse calvaria. In addition, treatment of calvaria with isoprenaline or norepinephrine increased IL-6 synthesis in the organ culture system. These results indicate that in vivo adrenergic stimulation modulates the osteoblastic activity in mouse calvaria via noradrenergic nerve terminals. PMID- 12900384 TI - Flow-dependent regulation of endothelial nitric oxide synthase: role of protein kinases. AB - Vascular endothelial cells are directly and continuously exposed to fluid shear stress generated by blood flow. Shear stress regulates endothelial structure and function by controlling expression of mechanosensitive genes and production of vasoactive factors such as nitric oxide (NO). Though it is well known that shear stress stimulates NO production from endothelial nitric oxide synthase (eNOS), the underlying molecular mechanisms remain unclear and controversial. Shear induced production of NO involves Ca2+/calmodulin-independent mechanisms, including phosphorylation of eNOS at several sites and its interaction with other proteins, including caveolin and heat shock protein-90. There have been conflicting results as to which protein kinases-protein kinase A, protein kinase B (Akt), other Ser/Thr protein kinases, or tyrosine kinases-are responsible for shear-dependent eNOS regulation. The functional significance of each phosphorylation site is still unclear. We have attempted to summarize the current status of understanding in shear-dependent eNOS regulation. PMID- 12900385 TI - Rho's role in cell volume: sensing, strutting, or signaling? Focus on "Hyperosmotic stress activates Rho: differential involvement in Rho kinase dependent MLC phosphorylation and NKCC activation". PMID- 12900386 TI - PKC zeta participates in activation of inflammatory response induced by enteropathogenic E. coli. AB - We showed previously that enteropathogenic Escherichia coli (EPEC) infection of intestinal epithelial cells induces inflammation by activating NF-kappa B and upregulating IL-8 expression. We also reported that extracellular signal regulated kinases (ERKs) participate in EPEC-induced NF-kappa B activation but that other signaling molecules such as PKC zeta may be involved. The aim of this study was to determine whether PKC zeta is activated by EPEC and to investigate whether it also plays a role in EPEC-associated inflammation. EPEC infection induced the translocation of PKC zeta from the cytosol to the membrane and its activation as determined by kinase activity assays. Inhibition of PKC zeta by the pharmacological inhibitor rottlerin, the inhibitory myristoylated PKC zeta pseudosubstrate (MYR-PKC zeta-PS), or transient expression of a nonfunctional PKC zeta significantly suppressed EPEC-induced I kappa B alpha phosphorylation. Although PKC zeta can activate ERK, MYR-PKC zeta-PS had no effect on EPEC-induced stimulation of this pathway, suggesting that they are independent events. PKC zeta can regulate NF-kappa B activation by interacting with and activating I kappa B kinase (IKK). Coimmunoprecipitation studies showed that the association of PKC zeta and IKK increased threefold 60 min after infection. Kinase activity assays using immunoprecipitated PKC zeta-IKK complexes from infected intestinal epithelial cells and recombinant I kappa B alpha as a substrate showed a 2.5-fold increase in I kappa B alpha phosphorylation. PKC zeta can also regulate NF-kappa B by serine phosphorylation of the p65 subunit. Serine phosphorylation of p65 was increased after EPEC infection but could not be consistently attenuated by MYR PKC zeta-PS, suggesting that other signaling events may be involved in this particular arm of NF-kappa B regulation. We speculate that EPEC infection of intestinal epithelial cells activates several signaling pathways including PKC zeta and ERK that lead to NF-kappa B activation, thus ensuring the proinflammatory response. PMID- 12900387 TI - Effects of nonselective cation channels and PI3K on endothelin-1-induced PYK2 tyrosine phosphorylation in C6 glioma cells. AB - We recently demonstrated that endothelin-1 (ET-1) activates two types of Ca2+ permeable nonselective cation channels (designated NSCC-1 and NSCC-2) in C6 glioma cells. In the present study, we investigated the effects of NSCCs on the ET-1-induced proline-rich tyrosine kinase 2 (PYK2) phosphorylation in C6 glioma cells. In addition, we examined the effects of phosphoinositide 3-kinase (PI3K) on the ET-1-induced NSCCs activation and PYK2 phosphorylation. The PI3K inhibitors wortmannin and LY-294002 inhibited ET-1-induced Ca2+ influx through NSCC-2 but not NSCC-1. On the other hand, addition of these inhibitors after stimulation with ET-1 failed to suppress Ca2+ influx through NSCC-2. PYK2 phosphorylation was abolished by blocking Ca2+ influx through NSCCs. The PI3K inhibitors blocked the NSCC-2-dependent part of ET-1-induced PYK2 phosphorylation. These results indicate that 1) NSCC-2 is stimulated by ET-1 via a PI3K-dependent cascade, whereas NSCC-1 is stimulated via a PI3K-independent cascade; 2) PI3K seems to be required for the activation of the Ca2+ entry, but not for its maintenance; 3) Ca2+ influx through NSCC-1 and NSCC-2 plays an essential role in ET-1-induced PYK2 phosphorylation; and 4) PI3K is involved in the ET-1-induced PYK2 phosphorylation that depends on the Ca2+ influx through NSCC-2. PMID- 12900388 TI - In vitro and in vivo characterization of the minimal promoter region of the human thiamin transporter SLC19A2. AB - The molecular mechanisms involved in the regulation of thiamin transport in mammalian cells are poorly understood. Previous studies established that a human thiamin transporter, SLC19A2, plays a role in thiamin uptake in human tissues. We cloned the 5' regulatory region of the SLC19A2 gene, identified the minimal promoter required for basal activity, and located multiple putative cis elements. To further characterize the SLC19A2 promoter, we investigated, in the present study, the role of the putative cis elements in regulating the activity of the SLC19A2 promoter in vitro and confirmed the activity of the SLC19A2 promoter in vivo. In vitro studies demonstrated that mutation of specific cis elements in the SLC19A2 minimal promoter [Gut-enriched Krupple-like factor (GKLF), nuclear factor 1 (NF-1), and stimulating protein-1 (SP-1)] led to a decrease in activity. Using electrophoretic mobility shift assays, four specific DNA/protein complexes were identified. The interacting factors were determined by oligonucleotide competition and antibody supershift analysis and shown to be GKLF, NF-1, and SP 1. Cotransfection studies of the SLC19A2 promoter with an SP-1 containing vector in Drosophila SL2 cells further confirmed a role for SP-1 in regulating SLC19A2 promoter activity. In vivo studies using transgenic mice established the functionality of the full-length and minimal SLC19A2 promoters. Furthermore, our studies revealed that the pattern of expression of the SLC19A2 promoter Luciferase constructs in transgenic mice was similar to the reported SLC19A2 RNA expression pattern in native human tissues. The results demonstrate the importance of GKLF, NF-1, and SP-1 in regulating the activity of the SLC19A2 promoter and provide direct in vivo confirmation of promoter activity. PMID- 12900389 TI - Loss of active MEK1-ERK1/2 restores epithelial phenotype and morphogenesis in transdifferentiated MDCK cells. AB - Constitutive activation of the MAPK/ERK kinase (MEK)1-ERK2 signaling module in Madin-Darby canine kidney (MDCK)-C7 cells disrupts their ability to form cyst like structures in collagen gels and induces an invasive, myofibroblast-like phenotype. However, the reversibility of these cellular events, as well as the relative role of both MEK isoforms (MEK1 and MEK2) and both ERK isoforms (ERK1 and ERK2) during these processes, has not yet been investigated. We now report that loss of constitutively active MEK1 (caMEK1) and, thus, loss of active ERK1/2 in C7caMEK1 cells is associated with increased MEK2 protein expression, reexpression of ERK1 protein, and epithelial redifferentiation of these cells. The morphological changes toward an epithelial phenotype in these revertant cell lines (C7rev4, C7rev5, C7rev7) are reflected by the upregulation of epithelial marker proteins, such as E-cadherin, beta-catenin, and cytokeratin, by the loss of alpha-smooth muscle actin expression, and by the ability of these epithelial revertants to form well-organized spherical cysts when grown in three-dimensional collagen gels. Further evidence for a role of the MEK1-ERK1/2 module in epithelial-mesenchymal transition was obtained from the analysis of two novel, spontaneously transdifferentiated MDCK-C7 cell clones (C7e1 and C7e2 cells). In these clones, increased MEK1/2-ERK1/2 phosphorylation, reduced MEK2 protein expression, and loss of ERK1 protein expression is associated with phenotypic alterations similar to those observed in transdifferentiated C7caMEK1 cells. C7e1 cells at least partially regained some of their epithelial characteristics at higher passages. In contrast, C7e2 cells maintained a transdifferentiated phenotype at high passage, were unable to generate cyst-like epithelial structures, and retained invasive properties when grown on a three-dimensional collagen matrix. We conclude that in renal epithelial MDCK-C7 cells, stable epithelial-to-mesenchymal transition (EMT) is associated with loss of ERK1 protein expression, reduced MEK2 protein expression, and increased basal ERK2 phosphorylation. In contrast, loss of active MEK1-ERK1/2 results in increased MEK2 protein expression and reexpression of ERK1 protein, concomitant with the restoration of epithelial phenotype and the ability to form cystic structures. PMID- 12900390 TI - After Hrs with HIV. AB - To efficiently bud off from infected cells, HIV and other enveloped viruses hijack the host cellular machinery that is normally involved in vacuolar protein sorting and multivesicular body (MVB) biogenesis. The HIV Gag protein mimics hepatocyte growth factor-regulated tyrosine kinase substrate (Hrs), a modular adaptor protein that links membrane cargo recognition to its degradation after delivery to MVBs. In contrast to T cells, where HIV budding occurs at the plasma membrane, virus buds into vacuoles of macrophages, a process that may facilitate its spread within the infected host. PMID- 12900391 TI - Direct observation of microtubule dynamics at kinetochores in Xenopus extract spindles: implications for spindle mechanics. AB - Microtubule plus ends dynamically attach to kinetochores on mitotic chromosomes. We directly imaged this dynamic interface using high resolution fluorescent speckle microscopy and direct labeling of kinetochores in Xenopus extract spindles. During metaphase, kinetochores were stationary and under tension while plus end polymerization and poleward microtubule flux (flux) occurred at velocities varying from 1.5-2.5 micro m/min. Because kinetochore microtubules polymerize at metaphase kinetochores, the primary source of kinetochore tension must be the spindle forces that produce flux and not a kinetochore-based mechanism. We infer that the kinetochore resists translocation of kinetochore microtubules through their attachment sites, and that the polymerization state of the kinetochore acts a "slip-clutch" mechanism that prevents detachment at high tension. At anaphase onset, kinetochores switched to depolymerization of microtubule plus ends, resulting in chromosome-to-pole rates transiently greater than flux. Kinetochores switched from persistent depolymerization to persistent polymerization and back again during anaphase, bistability exhibited by kinetochores in vertebrate tissue cells. These results provide the most complete description of spindle microtubule poleward flux to date, with important implications for the microtubule-kinetochore interface and for how flux regulates kinetochore function. PMID- 12900392 TI - Microtubules are the only structural constituent of the spindle apparatus required for induction of cell cleavage. AB - Structural constituents of the spindle apparatus essential for cleavage induction remain undefined. Findings from various cell types using different approaches suggest the importance of all structural constituents, including asters, the central spindle, and chromosomes. In this study, we systematically dissected the role of each constituent in cleavage induction in grasshopper spermatocytes and narrowed the essential one down to bundled microtubules. Using micromanipulation, we produced "cells" containing only asters, a truncated central spindle lacking both asters and chromosomes, or microtubules alone. We show that furrow induction occurs under all circumstances, so long as sufficient microtubules are present. Microtubules, as the only spindle structural constituent, undergo dramatic, stage specific reorganizations, radiating toward cell cortex in "metaphase," disassembling in "anaphase," and bundling into arrays in "telophase." Furrow induction usually occurs at multisites around microtubule bundles, but only those induced by sustained bundles ingress. We suggest that microtubules, regardless of source, are the only structural constituent of the spindle apparatus essential for cleavage furrow induction. PMID- 12900393 TI - Vps27 recruits ESCRT machinery to endosomes during MVB sorting. AB - Down-regulation (degradation) of cell surface proteins within the lysosomal lumen depends on the function of the multivesicular body (MVB) sorting pathway. The function of this pathway requires the class E vacuolar protein sorting (Vps) proteins. Of the class E Vps proteins, both the ESCRT-I complex (composed of the class E proteins Vps23, 28, and 37) and Vps27 (mammalian hepatocyte receptor tyrosine kinase substrate, Hrs) have been shown to interact with ubiquitin, a signal for entry into the MVB pathway. We demonstrate that activation of the MVB sorting reaction is dictated largely through interactions between Vps27 and the endosomally enriched lipid species phosphatidylinositol 3-phosphate via the FYVE domain (Fab1, YGL023, Vps27, and EEA1) of Vps27. ESCRT-I then physically binds to Vps27 on endosomal membranes via a domain within the COOH terminus of Vps27. A peptide sequence in this domain, PTVP, is involved in the function of Vps27 in the MVB pathway, the efficient endosomal recruitment of ESCRT-I, and is related to a motif in HIV-1 Gag protein that is capable of interacting with Tsg101, the mammalian homologue of Vps23. We propose that compartmental specificity for the MVB sorting reaction is the result of interactions of Vps27 with phosphatidylinositol 3-phosphate and ubiquitin. Vps27 subsequently recruits/activates ESCRT-I on endosomes, thereby facilitating sorting of ubiquitinated MVB cargoes. PMID- 12900395 TI - Hrs regulates multivesicular body formation via ESCRT recruitment to endosomes. AB - Hrs and the endosomal sorting complexes required for transport, ESCRT-I, -II, and -III, are involved in the endosomal sorting of membrane proteins into multivesicular bodies and lysosomes or vacuoles. The ESCRT complexes are also required for formation of intraluminal endosomal vesicles and for budding of certain enveloped RNA viruses such as HIV. Here, we show that Hrs binds to the ESCRT-I subunit Tsg101 via a PSAP motif that is conserved in Tsg101-binding viral proteins. Depletion of Hrs causes a reduction in membrane-associated ESCRT-I subunits, a decreased number of multivesicular bodies and an increased size of late endosomes. Even though Hrs mainly localizes to early endosomes and Tsg101 to late endosomes, the two proteins colocalize on a subpopulation of endosomes that contain lyso-bisphosphatidic acid. Overexpression of Hrs causes accumulation of Tsg101 on early endosomes and prevents its localization to late endosomes. We conclude that Hrs mediates the initial recruitment of ESCRT-I to endosomes and, thereby, indirectly regulates multivesicular body formation. PMID- 12900394 TI - HIV Gag mimics the Tsg101-recruiting activity of the human Hrs protein. AB - The HIV-1 Gag protein recruits the cellular factor Tsg101 to facilitate the final stages of virus budding. A conserved P(S/T)AP tetrapeptide motif within Gag (the "late domain") binds directly to the NH2-terminal ubiquitin E2 variant (UEV) domain of Tsg101. In the cell, Tsg101 is required for biogenesis of vesicles that bud into the lumen of late endosomal compartments called multivesicular bodies (MVBs). However, the mechanism by which Tsg101 is recruited from the cytoplasm onto the endosomal membrane has not been known. Now, we report that Tsg101 binds the COOH-terminal region of the endosomal protein hepatocyte growth factor regulated tyrosine kinase substrate (Hrs; residues 222-777). This interaction is mediated, in part, by binding of the Tsg101 UEV domain to the Hrs 348PSAP351 motif. Importantly, Hrs222-777 can recruit Tsg101 and rescue the budding of virus like Gag particles that are missing native late domains. These observations indicate that Hrs normally functions to recruit Tsg101 to the endosomal membrane. HIV-1 Gag apparently mimics this Hrs activity, and thereby usurps Tsg101 and other components of the MVB vesicle fission machinery to facilitate viral budding. PMID- 12900396 TI - A mutant heterodimeric myosin with one inactive head generates maximal displacement. AB - Each of the heads of the motor protein myosin II is capable of supporting motion. A previous report showed that double-headed myosin generates twice the displacement of single-headed myosin (Tyska, M.J., D.E. Dupuis, W.H. Guilford, J.B. Patlak, G.S. Waller, K.M. Trybus, D.M. Warshaw, and S. Lowey. 1999. Proc. Natl. Acad. Sci. USA. 96:4402-4407). To determine the role of the second head, we expressed a smooth muscle heterodimeric heavy meromyosin (HMM) with one wild-type head, and the other locked in a weak actin-binding state by introducing a point mutation in switch II (E470A). Homodimeric E470A HMM did not support in vitro motility, and only slowly hydrolyzed MgATP. Optical trap measurements revealed that the heterodimer generated unitary displacements of 10.4 nm, strikingly similar to wild-type HMM (10.2 nm) and approximately twice that of single-headed subfragment-1 (4.4 nm). These data show that a double-headed molecule can achieve a working stroke of approximately 10 nm with only one active head and an inactive weak-binding partner. We propose that the second head optimizes the orientation and/or stabilizes the structure of the motion-generating head, thereby resulting in maximum displacement. PMID- 12900397 TI - Local ERM activation and dynamic growth cones at Schwann cell tips implicated in efficient formation of nodes of Ranvier. AB - Nodes of Ranvier are specialized, highly polarized axonal domains crucial to the propagation of saltatory action potentials. In the peripheral nervous system, axo glial cell contacts have been implicated in Schwann cell (SC) differentiation and formation of the nodes of Ranvier. SC microvilli establish axonal contact at mature nodes, and their components have been observed to localize early to sites of developing nodes. However, a role for these contacts in node formation remains controversial. Using a myelinating explant culture system, we have observed that SCs reorganize and polarize microvillar components, such as the ezrin-binding phosphoprotein 50 kD/regulatory cofactor of the sodium-hydrogen exchanger isoform 3 (NHERF-1), actin, and the activated ezrin, radixin, and moesin family proteins before myelination in response to inductive signals. These components are targeted to the SC distal tips where live cell imaging reveals novel, dynamic growth cone-like behavior. Furthermore, localized activation of the Rho signaling pathway at SC tips gives rise to these microvillar component-enriched "caps" and influences the efficiency of node formation. PMID- 12900398 TI - Dynamic changes in the osteoclast cytoskeleton in response to growth factors and cell attachment are controlled by beta3 integrin. AB - The beta3 integrin cytoplasmic domain, and specifically S752, is critical for integrin localization and osteoclast (OC) function. Because growth factors such as macrophage colony-stimulating factor and hepatocyte growth factor affect integrin activation and function via inside-out signaling, a process requiring the beta integrin cytoplasmic tail, we examined the effect of these growth factors on OC precursors. To this end, we retrovirally expressed various beta3 integrins with cytoplasmic tail mutations in beta3-deficient OC precursors. We find that S752 in the beta3 cytoplasmic tail is required for growth factor induced integrin activation, cytoskeletal reorganization, and membrane protrusion, thereby affecting OC adhesion, migration, and bone resorption. The small GTPases Rho and Rac mediate cytoskeletal reorganization, and activation of each is defective in OC precursors lacking a functional beta3 subunit. Activation of the upstream mediators c-Src and c-Cbl is also dependent on beta3. Interestingly, although the FAK-related kinase Pyk2 interacts with c-Src and c Cbl, its activation is not disrupted in the absence of functional beta3. Instead, its activation is dependent upon intracellular calcium, and on the beta2 integrin. Thus, the beta3 cytoplasmic domain is responsible for activation of specific intracellular signals leading to cytoskeletal reorganization critical for OC function. PMID- 12900399 TI - Regulation of vascular tone in animals overexpressing the sarcolemmal calcium pump. AB - The mechanisms governing vascular smooth muscle tone are incompletely understood. In particular, the role of the sarcolemmal calcium pump PMCA (plasma membrane calmodulin-dependent calcium ATPase), which extrudes Ca2+ from the cytosol, and its importance compared with the sodium/calcium exchanger remain speculative. To test whether the PMCA is a regulator of vascular tone, we generated transgenic mice overexpressing the human PMCA4b under control of the arterial smooth muscle specific SM22alpha promoter. This resulted in an elevated systolic blood pressure compared with littermate controls. In PMCA-overexpressing mice, endothelium dependent relaxation of norepinephrine-preconstricted aortic rings to acetylcholine did not differ from wild type controls (76 +/- 8% versus 79 +/- 8% of maximum relaxation; n = 12, n.s.). De-endothelialized aortas of transgenic mice exhibited stronger maximum contraction to KCl (100 mmol/liter) compared with controls (86 +/- 6% versus 68 +/- 7% of reference KCl contraction at the beginning of the experiment; p <0.05). Preincubation of de-endothelialized vessels with the nitric oxide synthase (NOS) inhibitor l-NAME (l-N(G) nitroarginine methyl ester) (10-5 mol/liter) resulted in a stronger contraction to KCl (p <0.05 versus without l-NAME), thus unmasking vasodilatory effects of inherent NO production. Maximum contraction to KCl after preincubation with l NAME did not differ between PMCA mice and controls. In analogy to the results in PMCA-overexpressing mice, contractions of de-endothelialized aortas of neuronal NOS-deficient mice to KCl were significantly increased compared with controls (151 +/- 5% versus 131 +/- 6% of reference KCl contraction; p <0.05). In conclusion, our data suggest a model in which the sarcolemmal Ca2+ pump down regulates activity of the vascular smooth muscle Ca2+/calmodulin-dependent neuronal NOS by a functionally relevant interaction. Therefore, the PMCA represents a novel regulator of vascular tone. PMID- 12900400 TI - Enhanced expression of L-type Cav1.3 calcium channels in murine embryonic hearts from Cav1.2-deficient mice. AB - Voltage-gated calcium (Ca2+) channels play a key role in the control of heart contraction and are essential for normal heart development. The Cav1.2 L-type calcium channel is the predominant isoform in cardiomyocytes and is essential for excitation-contraction coupling. Although the inactivation of the Cav1.2 gene caused embryonic lethality before embryonic day E14.5, hearts were contracting before E14 depending on a dihydropyridine-sensitive calcium influx. We analyzed the consequences of the deletion of the Cav1.2 channel on the expression level of other voltage-gated calcium channels in the embryonic mouse heart and isolated cardiomyocytes. A strong compensatory up-regulation of the Cav1.3 calcium channel was observed on the mRNA as well as on the protein level. Reverse transcriptase PCR indicated that the recently identified new Cav1.3(1b) isoform was strongly up regulated, whereas a more moderate increase was found for the Cav1.3(1a) variant. Heterologous expression of Cav1.3(1b) in HEK293 cells induced Ba2+ currents with properties similar to those found in Cav1.2 (-/-) cardiomyocytes, suggesting that this isoform constitutes a major component of the residual L-type calcium current in Cav1.2 (-/-) cardiomyocytes. In summary, our results imply that calcium channel expression is dynamically regulated during heart development and that the Cav1.3 channel may substitute for Cav1.2 during early embryogenesis. PMID- 12900401 TI - Characterization of the interaction between neuronal RNA-binding protein HuD and AU-rich RNA. AB - Hu proteins have been shown to bind to AU-rich elements (AREs) in the 3' untranslated region of unstable mRNAs. They can thereby inhibit the decay of labile transcripts by antagonizing destabilizing proteins that target these AU rich sequences. Here we examine the sequence preferences of HuD to elucidate its possible role in counteracting mRNA decay. Using repeats of the prototype destabilizing sequence UU(AUUU)nAUU, we show that all three HuD RNA-binding domains participate in binding to AU-tracts that can be as short as 13 residues, depending on the position of the remaining As. Removal of the A residues, resulting in a poly(U)-tract, increased the affinity of HuD for RNA, suggesting that the presence of As in destabilizing elements might favor the recruitment of other proteins and/or prevent HuD from binding too tightly to AREs. In vitro selection experiments with randomized RNAs confirmed the preference of HuD for poly(U). RNA binding analysis of the related protein HuB showed a similar preference for poly(U). In contrast, tristetraprolin, an mRNA destabilizing protein, strongly prefers AU-rich RNA. Many labile mRNAs contain U-tracts in or near their AREs. Individual AREs may thus differentially affect mRNA half-life by recruiting a unique complement of stabilizing and destabilizing factors. PMID- 12900402 TI - Direct activation of phospholipase C-epsilon by Rho. AB - Unique among the phospholipase C isozymes, the recently identified phospholipase C-epsilon (PLC-epsilon) contains an amino-terminal CDC25 domain capable of catalyzing nucleotide exchange on Ras family GTPases as well as a tandem array of Ras-associating (RA) domains near its carboxyl terminus that are effector binding sites for activated H-Ras and Rap. To determine whether other small GTPases activate PLC-epsilon, we measured inositol phosphate accumulation in COS-7 cells expressing a broad range of GTPase-deficient mutants of Ras superfamily proteins. RhoA, RhoB, and RhoC all markedly stimulated inositol phosphate accumulation in PLC-epsilon-expressing cells. This stimulation matched or exceeded phospholipase activation promoted by co-expression of PLC-epsilon with the known regulators Ras, Galpha12/13, or Gbeta1gamma2. In contrast, little effect was observed with the other Rho family members Rac1, Rac2, Rac3, and Cdc42. Truncation of the two carboxyl-terminal RA domains caused loss of responsiveness to H-Ras but not to Rho. Truncation of PLC-epsilon to remove the CDC25 and pleckstrin homology (PH) domains also did not cause loss of responsiveness to Rho, Galpha12/13, or Gbeta1gamma2. Comparative sequence analysis of mammalian phospholipase C isozymes revealed a unique approximately 65 amino acid insert within the catalytic core of PLC-epsilon not present in PLC-beta, gamma, delta, or zeta. A PLC-epsilon construct lacking this region was no longer activated by Rho or Galpha12/13 but retained regulation by Gbetagamma and H-Ras. GTP-dependent interaction of Rho with PLC-epsilon was illustrated in pull-down experiments with GST-Rho, and this interaction was retained in the PLC-epsilon construct lacking the unique insert within the catalytic core. These results are consistent with the conclusion that Rho family GTPases directly interact with PLC-epsilon by a mechanism independent of the CDC25 or RA domains. A unique insert within the catalytic core of PLC epsilon imparts responsiveness to Rho, which may signal downstream of Galpha12/13 in the regulation of PLC-epsilon, because activation by both Rho and Galpha12/13 is lost in the absence of this sequence. PMID- 12900403 TI - Molecular cloning, expression, and characterization of novel hemolytic lectins from the mushroom Laetiporus sulphureus, which show homology to bacterial toxins. AB - We describe herein the cDNA cloning, expression, and characterization of a hemolytic lectin and its related species from the parasitic mushroom Laetiporus sulphureus. The lectin designated LSL (L. sulphureus lectin), is a tetramer composed of subunits of approximately 35 kDa associated by non-covalent bonds. From a cDNA library, three similar full-length cDNAs, termed LSLa, LSLb, and LSLc, were generated, each of which had an open reading frame of 945 bp encoding 315 amino acid residues. These proteins share 80-90% sequence identity and showed structural similarity to bacterial toxins: mosquitocidal toxin (MTX2) from Bacillus sphaericus and alpha toxin from Clostridium septicum. Native and recombinant forms of LSL showed hemagglutination and hemolytic activity and both activities were inhibited by N-acetyllactosamine, whereas a C-terminal deletion mutant of LSLa (LSLa-D1) retained hemagglutination, but not hemolytic activity, indicating the N-terminal domain is a carbohydrate recognition domain and the C terminal domain functions as an oligomerization domain. The LSL-mediated hemolysis was protected osmotically by polyethylene glycol 4000 and maltohexaose. Inhibition studies showed that lacto-N-neotetraose (Galbeta1-4GlcNAcbeta1 3Galbeta1-4Glc) was the best inhibitor for LSL. These results indicate that LSL is a novel pore-forming lectin homologous to bacterial toxins. PMID- 12900404 TI - Palmitoylation of the V2 vasopressin receptor carboxyl tail enhances beta arrestin recruitment leading to efficient receptor endocytosis and ERK1/2 activation. AB - A large number of G protein-coupled receptors are palmitoylated on cysteine residues located in their carboxyl tail, but the general role of this post translational modification remains poorly understood. Here we show that preventing palmitoylation of the V2 vasopressin receptor, by site-directed mutagenesis of cysteines 341 and 342, significantly delayed and decreased both agonist-promoted receptor endocytosis and mitogen-activated protein kinase activation. Pharmacological blockade of receptor endocytosis is without effect on the vasopressin-stimulated mitogen-activated protein kinase activity, excluding the possibility that the reduced kinase activation mediated by the palmitoylation less mutant could result from altered receptor endocytosis. In contrast, two dominant negative mutants of beta-arrestin which inhibit receptor endocytosis also attenuated vasopressin-stimulated mitogen-activated protein kinase activity, suggesting that the scaffolding protein, beta-arrestin, represents the common link among receptor palmitoylation, endocytosis, and kinase activation. Coimmunoprecipitation and bioluminescence resonance energy transfer experiments confirmed that inhibiting receptor palmitoylation considerably reduced the vasopressin-stimulated recruitment of beta-arrestin to the receptor. Interestingly, the changes in beta-arrestin recruitment kinetics were similar to those observed for vasopressin-stimulated receptor endocytosis and mitogen activated protein kinase activation. Taken together the results indicate that palmitoylation enhances the recruitment of beta-arrestin to the activated V2 vasopressin receptor thus facilitating processes requiring the scaffolding action of beta-arrestin. PMID- 12900405 TI - Kinetics of the superoxide reductase catalytic cycle. AB - The steady state kinetics of a Desulfovibrio (D.) vulgaris superoxide reductase (SOR) turnover cycle, in which superoxide is catalytically reduced to hydrogen peroxide at a [Fe(His)4(Cys)] active site, are reported. A proximal electron donor, rubredoxin, was used to supply reducing equivalents from NADPH via ferredoxin: NADP+ oxidoreductase, and xanthine/xanthine oxidase was used to provide a calibrated flux of superoxide. SOR turnover in this system was well coupled, i.e. approximately 2O*2 reduced:NADPH oxidized over a 10-fold range of superoxide flux. The reduction of the ferric SOR active site by reduced rubredoxin was independently measured to have a second-order rate constant of approximately 1 x 10(6) m-1 s-1. Analysis of the kinetics showed that: (i) 1 microM SOR can convert a 10 microM/min superoxide flux to a steady state superoxide concentration of 10(-10) m, during which SOR turns over about once every 6 s, (ii) the diffusion-controlled reaction of reduced SOR with superoxide is the slowest process during turnover, and (iii) neither ligation nor deligation of the active site carboxylate of SOR limits the turnover rate. An intracellular SOR concentration on the order of 10 microM is estimated to be the minimum required for lowering superoxide to sublethal levels in aerobically growing SOD knockout mutants of Escherichia coli. SORs from Desulfovibrio gigas and Treponema pallidum showed similar turnover rates when substituted for the D. vulgaris SOR, whereas superoxide dismutases showed no SOR activity in our assay. These results provide quantitative support for previous suggestions that, in times of oxidative stress, SORs efficiently divert intracellular reducing equivalents to superoxide. PMID- 12900406 TI - Structural and functional uncoupling of the enzymatic and angiogenic inhibitory activities of tissue inhibitor of metalloproteinase-2 (TIMP-2): loop 6 is a novel angiogenesis inhibitor. AB - Tissue inhibitors of metalloproteinases (TIMPs) regulate tumor growth, progression, and angiogenesis in a variety of experimental cancer models and in human malignancies. Results from numerous studies have revealed important differences between TIMP family members in their ability to inhibit angiogenic processes in vitro and angiogenesis in vivo despite their universal ability to inhibit matrix metalloproteinase (MMP) activity. To address these differences, a series of structure-function studies were conducted to identify and to characterize the anti-angiogenic domains of TIMP-2, the endogenous MMP inhibitor that uniquely inhibits capillary endothelial cell (EC) proliferation as well as angiogenesis in vivo. We demonstrate that the COOH-terminal domain of TIMP-2 (T2C) inhibits the proliferation of capillary EC at molar concentrations comparable with those previously reported for intact TIMP-2, while the NH2 terminal domain (T2N), which inhibits MMP activity, has no significant anti proliferative effect. Interestingly, although both T2N and T2C inhibited embryonic angiogenesis, only T2C resulted in the potent inhibition of angiogenesis driven by the exogenous addition of angiogenic mitogen, suggesting that MMP inhibition alone may not be sufficient to inhibit the aggressive neovascularization characteristic of aberrant angiogenesis. We further mapped the anti-proliferative activity of T2C to a 24-amino acid peptide corresponding to Loop 6 of TIMP-2 and show that Loop 6 is a potent inhibitor of both embryonic and mitogen-stimulated angiogenesis in vivo. These findings demonstrate that TIMP-2 possesses two distinct types of anti-angiogenic activities which can be uncoupled from each other, the first represented by its MMP-dependent inhibitory activity which can inhibit only embryonic neovascularization and the second represented by an MMP-independent activity which inhibits both normal angiogenesis and mitogen driven angiogenesis in vivo. In addition, we report, for the first time, the discovery of Loop 6 as a novel and potent inhibitor of angiogenesis. PMID- 12900407 TI - AMP-activated protein kinase activity is critical for hypoxia-inducible factor-1 transcriptional activity and its target gene expression under hypoxic conditions in DU145 cells. AB - AMP-activated protein kinase (AMPK) functions as an energy sensor to provide metabolic adaptations under the ATP-deprived conditions such as hypoxia. In the present study, we considered a role of AMPK in the adaptive response to hypoxia by examining whether AMPK is involved in the regulation of hypoxia-inducible factor-1 (HIF-1), a heterodimeric transcription factor that is critical for hypoxic induction of physiologically important genes. We demonstrate that hypoxia or CoCl2 rapidly activated AMPK in DU145 human prostate cancer cells, and its activation preceded the induction of HIF-1 alpha expression. Under these conditions, blockade of AMPK activity by a pharmacological or molecular approach significantly attenuated hypoxia-induced responses such as HIF-1 target gene expression, secretion of vascular endothelial growth factor, glucose uptake, and HIF-1-dependent reporter gene expression, indicating that AMPK is critical for the HIF-1 transcriptional activity and its target gene expression. Its functional requirement for HIF-1 activity was also demonstrated in several different cancer cell lines, but AMPK activation alone was not sufficient to stimulate the HIF-1 transcriptional activity. We further present data showing that AMPK transmits a positive signal for HIF-1 activity via a signaling pathway that is independent of phosphatidylinositol 3-kinase/AKT and several mitogen-activated protein kinases. Taken together, our results suggest that AMPK is a novel and critical component of HIF-1 regulation, implying its new roles in oxygen-regulated cellular phenomena. PMID- 12900408 TI - Tyrosine phosphorylation of the beta2 subunit of clathrin adaptor complex AP-2 reveals the role of a di-leucine motif in the epidermal growth factor receptor trafficking. AB - Tyrosine phosphorylation of the beta2 subunit of clathrin adaptor complex AP-2 was detected in three types of cells treated with epidermal growth factor (EGF). The tyrosine phosphorylation was observed during recruitment of EGF receptors into coated pits at 4 degrees C and reached maximum at 37 degrees C at post recruitment stages of endocytosis. An inhibitor of EGF receptor kinase completely abolished this phosphorylation in all cell types, whereas the inhibitor of Src family kinases partially inhibited beta2 phosphorylation in A-431 cells but not in HeLa cells. By using beta2 subunit tagged with yellow fluorescent protein that is effectively assembled into AP-2 complex, the major phosphorylation site of beta2 was mapped to Tyr-6. Analysis of cells expressing dominant-interfering mutant mu2 subunit of AP-2 suggested that beta2 phosphorylation is partially mediated by the receptor interaction with the mu2 subunit. Mutation of leucine residues 1010 and 1011 motif in the EGF receptor resulted in the severe inhibition of beta2 tyrosine phosphorylation. From these data, we propose that interactions of the EGF receptor with AP-2 mediated by the receptor 974YRAL and di-leucine motifs may contribute to beta2 tyrosine phosphorylation. Surprisingly, mutation of the Leu-1010/Leu-1011 motif resulted in impaired degradation of EGF receptors, suggesting the role of this motif in lysosomal targeting of the receptor. PMID- 12900409 TI - Akt-mediated cardiomyocyte survival pathways are compromised by G alpha q-induced phosphoinositide 4,5-bisphosphate depletion. AB - Expression of the wild type alpha subunit of Gq (GqWT) in cardiomyocytes induces hypertrophy, whereas a constitutively active G alpha q subunit (GqQ209L) induces apoptosis. Akt phosphorylation increases with GqWT expression but is markedly attenuated in cardiomyocytes expressing GqQ209L or in those expressing GqWT and treated with agonist. A membrane-targeted Akt rescues GqQ209L-expressing cardiomyocytes from apoptotic cell death. In contrast, leukemia inhibitory factor fails to activate Akt or promote cell survival in these cells. Association of Akt and PDK-1 with the membrane is also diminished in GqQ209L-expressing cardiomyocytes. Phosphatidylinositol 3,4,5-trisphosphate (PIP3), the primary regulator of Akt, increases significantly in GqWT-expressing cells but not in cardiomyocytes expressing GqQ209L. Levels of phosphatidylinositol 4,5 bisphosphate (PIP2), the immediate precursor of PIP3, are also markedly lower in GqQ209L-expressing compared to control cells. Expression of a GqQ209L mutant that has diminished capacity to activate phospholipase C does not decrease PIP2 or Akt or induce apoptosis. In transgenic mice with cardiac G alpha q overexpression, heart failure and increased cardiomyocyte apoptosis develop during the peripartal period. Akt phosphorylation and PIP2 levels decrease concomitantly. Our findings suggest that an Akt-mediated cell survival pathway is compromised by the diminished availability of PIP2 elicited by pathological levels of Gq activity. PMID- 12900410 TI - Ganglioside glycosyltransferases organize in distinct multienzyme complexes in CHO-K1 cells. AB - The synthesis of gangliosides is compartmentalized in the Golgi complex. In most cells, glycosylation of LacCer, GM3, and GD3 to form higher order species (GA2, GM2, GD2, GM1, GD1b) is displaced toward the most distal aspects of the Golgi and the trans-Golgi network, where the involved transferases (GalNAcT and GalT2) form physical and functional associations. Glycosylation of the simple species LacCer, GM3, and GD3, on the other hand, is displaced toward more proximal Golgi compartments, and we investigate here whether the involved transferases (GalT1, SialT1, and SialT2) share the property of forming physical associations. Co immunoprecipitation experiments from membranes of CHO-K1 cells expressing epitope tagged versions of these enzymes indicate that GalT1, SialT1, and SialT2 associate physically in a SialT1-dependent manner and that their N-terminal domains participate in these interactions. Microscopic fluorescence resonance energy transfer and fluorescence recovery after photobleaching in living cells confirmed the interactions, and in addition to showing a Golgi apparatus localization of the complexes, mapped their formation to the endoplasmic reticulum. Neither co-immunoprecipitation nor fluorescence resonance energy transfer detected interactions between either GalT2 or GalNAcT and GalT1 or SialT1 or SialT2. These results, and triple color imaging of Golgi-derived microvesicles in nocodazole-treated cells, suggest that ganglioside synthesis is organized in distinct units each formed by associations of particular glycosyltransferases, which concentrate in different sub-Golgi compartments. PMID- 12900411 TI - Amino acids 1-1,680 of ryanodine receptor type 1 hold critical determinants of skeletal type for excitation-contraction coupling. Role of divergence domain D2. AB - To identify domains of the ryanodine receptor (RyR1) that are functionally relevant for excitation-contraction (EC) coupling in vivo, we have studied the ability of RyR1/RyR3 chimera to rescue skeletal EC coupling in dyspedic myotubes. In this work we show that chimeric receptors containing amino acids 1-1,680 of RyR1 were able to render depolarization-induced Ca2+ release to RyR3. Within this region, residues 1,272-1,455, containing divergent domain D2 of RyR1, proved to be a critical element because the absence of this region selectively abolished depolarization-evoked Ca2+ transients without affecting chemically induced activation. Although the D2 domain by itself failed to restore skeletal EC coupling to RyR3, the addition of the D2 region resulted in a dramatic enhancement of EC coupling restored by an RyR3 chimera containing amino acids 1,681-3,770 of RyR1. These results suggest that although the D2 domain of RyR1 plays a key role during EC coupling, additional region(s) from the N-terminal end of RyR1 as well as previously identified regions of the central portion of the receptor are needed in order to allow normal EC coupling. PMID- 12900412 TI - p-Coumaroylnoradrenaline, a novel plant metabolite implicated in tomato defense against pathogens. AB - The Avr9 peptide elicitor from the fungus Cladosporium fulvum, the bacterial pathogen Pseudomonas syringae pathovar tomato carrying the avirulence gene avrPto (Pst (avrPto)), and the organophosphorous insecticide fenitrothion induce resistance-related responses in tomato lines carrying the Cf-9, Pto, and Fen genes, respectively. These responses were associated with synthesis of p coumaroyloctopamine and p-coumaroylnoradrenaline, a novel compound for plants. In susceptible near isogenic tomato lines (Cf-0, pto, fen) and wounded tomato leaves, the levels of these compounds were reduced or undetectable. The elevated levels of p-coumaroyloctopamine and p-coumaroylnoradrenaline were accompanied by elevated mRNA levels of genes encoding phenylalanine ammonia lyase, p-coumarate CoA ligase, and hydroxycinnamoyl-CoA:tyramine N-(hydroxycinnamoyl)transferase (THT), enzymes that are involved in the hydroxycinnamic acid amide biosynthesis. Southern hybridization indicated that THT is encoded by a multigene family in tomato. Four different THT full-length cDNAs were derived by reverse transcriptase-PCR using degenerate primers based on potato and tobacco THT sequences. Transcripts for all four homologs were present in unchallenged tomato leaves, but only tomTHT1-3 was highly expressed following challenge with Pst (avrPto). Furthermore, tomTHT1-3 showed a more substantial and rapid induction in the incompatible interaction than in the compatible interaction. The cDNAs tomTHT1-3, tomTHT7-1, and tomTHT7-8 encoded proteins with a high degree of amino acid sequence homology, although the recombinant proteins had different preferences for octopamine and noradrenaline. The fourth cDNA, tomTHT1-4, directed synthesis of a truncated enzymatically inactive protein due to the presence of a premature stop codon. PMID- 12900413 TI - IQGAP1 promotes cell motility and invasion. AB - The dynamic processes of cell migration and invasion are largely coordinated by Rho family GTPases. The scaffolding protein IQGAP1 binds to Cdc42, increasing the amount of active Cdc42 both in vitro and in cells. Here we show that overexpression of IQGAP1 in mammalian cells enhances cell migration in a Cdc42- and Rac1-dependent manner. Importantly, cell motility was significantly decreased both by knock down of endogenous IQGAP1 using small interfering RNA and by transfection of a dominant negative IQGAP1 construct, IQGAP1DeltaGRD. Cell invasion was similarly altered by manipulating intracellular IQGAP1 concentrations. Moreover, invasion mediated by constitutively active Cdc42 was attenuated by IQGAP1DeltaGRD. Thus, IQGAP1 has a fundamental role in cell motility and invasion. PMID- 12900414 TI - Ebony, a novel nonribosomal peptide synthetase for beta-alanine conjugation with biogenic amines in Drosophila. AB - Using Ebony protein either expressed in Escherichia coli or in Schneider S2 cells, we provide evidence for its substrate specificity and reaction mechanism. Ebony activates beta-alanine to aminoacyladenylate by an adenylation domain and covalently attaches it as a thioester to a thiolation domain in a nonribosomal peptide synthetase (NRPS) related mechanism. In a second reaction, biogenic amines act as external nucleophiles on beta-alanyl-S-pantetheine-Ebony, thereby releasing in a fast reaction the dipeptide (peptidoamine) in a process that is novel in higher eucaryotes. Therefore, we define Ebony as a beta-alanyl-biogenic amine synthetase. Insight into the reaction mechanism stems from mutational analysis of an invariant serine that disclosed Ebony as a multienzyme with functional analogy to the starting modules of NRPSs. In light of a putative biogenic amine-deactivating capacity, Ebony function in the nervous system must be reconsidered. We propose that in the Drosophila eye Ebony is involved in the transmission process by inactivation of histamine through beta-alanyl conjugation. PMID- 12900415 TI - Functional analysis of interleukin 6 response elements (IL-6REs) on the human gamma-fibrinogen promoter: binding of hepatic Stat3 correlates negatively with transactivation potential of type II IL-6REs. AB - Several families of transcription factors play important roles in modulating liver-specific gene expression during an acute phase response (APR). Stat3/APR factor is the main transactivator of gene expression by the interleukin (IL)-6 family of cytokines signaling through gp130. During an APR, fibrinogen (FBG) genes are coordinately up-regulated by IL-6 and glucocorticoids. Except for rat gammaFBG, attempts to demonstrate direct binding of IL-6-activated Stat3 to FBG CTGGGAA promoter elements have not been successful. Herein we show the presence of three functional type II IL-6 response elements (IL-6REs) on the human gammaFBG promoter and that the magnitude of Stat3 binding to these elements correlates negatively with their functional activity in reporter gene assays. Stat3-specific binding to gammaFBG IL-6REs was confirmed by cross-competition with alpha2-macroglobulin IL-6RE and specific interactions with anti-Stat3 in electrophoretic mobility shift assays. All type II IL-6REs contributed to full promoter activity; however, transactivation from Site II at -306 to -301 was strongest. In contrast to a previous report, IL-6 failed to induce activation of serum amyloid A-activating factor-1/c-Myc-associated zinc finger protein (SAF 1/MAZ), and mutation of the SAF-1RE had little effect on IL-6 induction of gammaFBG promoter activity. In the absence of a functional glucocorticoid receptor response element, dexamethasone potentiated IL-6-induced gammaFBG promoter activity 2-fold, requiring promoter-proximal Site I and Site II; the promoter-distal Site III had no effect on dexamethasone potentiation of IL-6 induced promoter activity. Notably the propensity for Stat3 binding to human gammaFBG IL-6REs was low compared with Stat3 binding to the alpha2-macroglobulin IL-6RE. Together these data suggest that Stat3 transactivation via IL-6REs on FBG promoters likely involves participation of additional transcription factors and/or coactivators to achieve optimal coordinated up-regulation during an APR. PMID- 12900417 TI - Cardiomyopathic tropomyosin mutations that increase thin filament Ca2+ sensitivity and tropomyosin N-domain flexibility. AB - The relationship between tropomyosin thermal stability and thin filament activation was explored using two N-domain mutants of alpha-striated muscle tropomyosin, A63V and K70T, each previously implicated in familial hypertrophic cardiomyopathy. Both mutations had prominent effects on tropomyosin thermal stability as monitored by circular dichroism. Wild type tropomyosin unfolded in two transitions, separated by 10 degrees C. The A63V and K70T mutations decreased the melting temperature of the more stable of these transitions by 4 and 10 degrees C, respectively, indicating destabilization of the N-domain in both cases. Global analysis of all three proteins indicated that the tropomyosin N domain and C-domain fold with a cooperative free energy of 1.0-1.5 kcal/mol. The two mutations increased the apparent affinity of the regulatory Ca2+ binding sites of thin filament in two settings: Ca2+-dependent sliding speed of unloaded thin filaments in vitro (at both pH 7.4 and 6.3), and Ca2+ activation of the thin filament-myosin S1 ATPase rate. Neither mutation had more than small effects on the maximal ATPase rate in the presence of saturating Ca2+ or on the maximal sliding speed. Despite the increased tropomyosin flexibility implied by destabilization of the N-domain, neither the cooperativity of thin filament activation by Ca2+ nor the cooperative binding of myosin S1-ADP to the thin filament was altered by the mutations. The combined results suggest that a more dynamic tropomyosin N-domain influences interactions with actin and/or troponin that modulate Ca2+ sensitivity, but has an unexpectedly small effect on cooperative changes in tropomyosin position on actin. PMID- 12900416 TI - UGT73C6 and UGT78D1, glycosyltransferases involved in flavonol glycoside biosynthesis in Arabidopsis thaliana. AB - Flavonol glycosides constitute one of the most prominent plant natural product classes that accumulate in the model plant Arabidopsis thaliana. To date there are no reports of functionally characterized flavonoid glycosyltransferases in Arabidopsis, despite intensive research efforts aimed at both flavonoids and Arabidopsis. In this study, flavonol glycosyltransferases were considered in a functional genomics approach aimed at revealing genes involved in determining the flavonol-glycoside profile. Candidate glycosyltransferase-encoding genes were selected based on homology to other known flavonoid glycosyltransferases and two T-DNA knockout lines lacking flavonol-3-O-rhamnoside-7-O-rhamnosides (ugt78D1) and quercetin-3-O-rhamnoside-7-O-glucoside (ugt73C6 and ugt78D1) were identified. To confirm the in planta results, cDNAs encoding both UGT78D1 and UGT73C6 were expressed in vitro and analyzed for their qualitative substrate specificity. UGT78D1 catalyzed the transfer of rhamnose from UDP-rhamnose to the 3-OH position of quercetin and kaempferol, whereas UGT73C6 catalyzed the transfer of glucose from UDP-glucose to the 7-OH position of kaempferol-3-O-rhamnoside and quercetin 3-O-rhamnoside, respectively. The present results suggest that UGT78D1 and UGT73C6 should be classified as UDP-rhamnose:flavonol-3-Orhamnosyltransferase and UDP-glucose:flavonol-3-O-glycoside-7-O-glucosyltransferase, respectively. PMID- 12900418 TI - Solution conformation of alphaA-conotoxin EIVA, a potent neuromuscular nicotinic acetylcholine receptor antagonist from Conus ermineus. AB - We report the solution three-dimensional structure of an alphaA-conotoxin EIVA determined by nuclear magnetic resonance spectroscopy and restrained molecular dynamics. The alphaA-conotoxin EIVA consists of 30 amino acids representing the largest peptide among the alpha/alphaA-family conotoxins discovered so far and targets the neuromuscular nicotinic acetylcholine receptor with high affinity. alphaA-Conotoxin EIVA consists of three distinct structural domains. The first domain is mainly composed of the Cys3-Cys11-disulfide loop and is structurally ill-defined with a large backbone root mean square deviation of 1.91 A. The second domain formed by residues His12-Hyp21 is extremely well defined with a backbone root mean square deviation of 0.52 A, thus forming a sturdy stem for the entire molecule. The third C-terminal domain formed by residues Hyp22-Gly29 shows an intermediate structural order having a backbone root mean square deviation of 1.04 A. A structurally ill-defined N-terminal first loop domain connected to a rigid central molecular stem seems to be the general structural feature of the alphaA-conotoxin subfamily. A detailed structural comparison between alphaA conotoxin EIVA and alphaA-conotoxin PIVA suggests that the higher receptor affinity of alphaA-conotoxin EIVA than alphaA-conotoxin PIVA might originate from different steric disposition and charge distribution in the second loop "handle" motif. PMID- 12900419 TI - Ca(2+)-dependent regulation of TrkB expression in neurons. AB - The neurotrophin brain-derived neurotrophic factor (BDNF), via activation of its receptor, tyrosine receptor kinase B (trkB), regulates a wide variety of cellular processes in the nervous system, including neuron survival and synaptic plasticity. Although the expression of BDNF is known to be Ca2+-dependent, the regulation of trkB expression has not been extensively studied. Here we report that depolarization of cultured mouse cortical neurons increased the expression of the full-length, catalytically active isoform of trkB without affecting expression of the truncated isoform. This increase in protein expression was accompanied by increased levels of transcripts encoding full-length, but not truncated, trkB. Depolarization also regulated transcription of the gene, TRKB, via entry of Ca2+ through voltage-gated Ca2+ channels and subsequent activation of Ca2+-responsive elements in the two TRKB promoters. Using transient transfection of neurons with TRKB promoter-luciferase constructs, we found that Ca2+ inhibited the upstream promoter P1 but activated the downstream promoter P2. Ca2+-dependent stimulation of TRKB expression requires two adjacent, non identical CRE sites located within P2. The coordinated regulation of BDNF and trkB by Ca2+ may play a role in activity-dependent survival and synaptic plasticity by enhancing BDNF signaling in electrically active neurons. PMID- 12900420 TI - Alleviating the suppression of glycogen synthase kinase-3beta by Akt leads to the phosphorylation of cAMP-response element-binding protein and its transactivation in intact cell nuclei. AB - Glycogen synthase kinase-3beta (GSK-3beta) activity is suppressed when it becomes phosphorylated on serine 9 by protein kinase B (Akt). To determine how GSK-3beta activity opposes Akt function we used various methods to alleviate GSK-3beta suppression in prostate carcinoma cells. In some experiments, LY294002, a specific inhibitor of phosphatidylinositol 3-kinase (a kinase involved in activating Akt) and tumor necrosis factor-alpha (TNF-alpha) were used to activate GSK-3beta. In other experiments mutant forms of GSK-3beta, GSK-3betadelta9 (a constitutively active deletion mutant of GSK-3beta) and GSK-3betaY216F (an inactive point mutant of GSK-3beta) were used to alter GSK-3beta activity. LY294002, TNF-alpha, and overexpression of wild-type GSK-3beta or of GSK 3betadelta9, but not GSK-3betaY216F, alleviated the suppression of GSK-3beta activity in prostate carcinoma cells and enhanced the turnover of beta-catenin. Forced expression of wild-type GSK-3beta or of GSK-3betadelta9, but not GSK 3betaY216F, suppressed cell growth and showed that the phosphorylation status of GSK-3beta can affect its intracellular distribution. When transcription factors activator protein-1 and cyclic AMP-response element (CRE)-binding protein were analyzed as targets of GSK-3beta activity, overexpression of wild-type GSK-3beta suppressed AP1-mediated transcription and activated CRE-mediated transcription. Overexpression of GSK-3betadelta9 caused an (80-fold) increase in CRE-mediated transcription, which was further amplified (up to 130-fold) by combining GSK 3betadelta9 overexpression with the suppression of Jun activity. This study also demonstrated for the first time that expression of constitutively active GSK 3betadelta9 results in the phosphorylation of CRE-binding protein on serine 129 and enhancement of CRE-mediated transcription in intact cell nuclei. PMID- 12900421 TI - Frameshifted beta-amyloid precursor protein (APP+1) is a secretory protein, and the level of APP+1 in cerebrospinal fluid is linked to Alzheimer pathology. AB - Molecular misreading of the beta-amyloid precursor protein (APP) gene generates mRNA with dinucleotide deletions in GAGAG motifs. The resulting truncated and partly frameshifted APP protein (APP+1) accumulates in the dystrophic neurites and the neurofibrillary tangles in the cortex and hippocampus of Alzheimer patients. In contrast, we show here that neuronal cells transfected with APP+1 proficiently secreted APP+1. Because various secretory APP isoforms are present in cerebrospinal fluid (CSF), this study aimed to determine whether APP+1 is also a secretory protein that can be detected in CSF. Post-mortem CSF was obtained at autopsy from 50 non-demented controls and 122 Alzheimer patients; all subjects were staged for neuropathology (Braak score). Unexpectedly, we found that the APP+1 level in the CSF of non-demented controls was much higher (1.75 ng/ml) than in the CSF of Alzheimer patients (0.51 ng/ml) (p < 0.001), and the level of APP+1 in CSF was inversely correlated with the severity of the neuropathology. Moreover the earliest neuropathological changes are already reflected in a significant decrease of the APP+1 level in CSF. These data show that APP+1 is normally secreted by neurons, preventing intra-neuronal accumulation of APP+1 in brains of non-demented controls without neurofibrillary pathology. PMID- 12900422 TI - Involvement of the VDE homing endonuclease and rapamycin in regulation of the Saccharomyces cerevisiae GSH11 gene encoding the high affinity glutathione transporter. AB - The Saccharomyces cerevisiae gene HGT1/GSH11 encodes the high affinity glutathione transporter and is repressed by cysteine added to the culture medium. It has been found previously that a 5'-upstream cis-element, CCGCCACAC, is responsible for regulating GSH11 expression and that several proteins bind to this element (Miyake, T., Kanayama, M., Sammoto, H., and Ono, B. (2002) Mol. Genet. Genomics 266, 1004-1011). In this report we present evidence that the most prominent of these proteins is VDE, known previously as the homing endonuclease encoded by VMA1. We show also that GSH11 is not expressed in a VDE-deleted strain and that inability to express the GSH11 of this strain is overcome by introduction of the coding region of VDE or the entire VMA1 gene. It is also found that VDE does not cut DNA in the vicinity of the GSH11 cis-element. Rapamycin, an inhibitor of the target of rapamycin (TOR) signal-transduction system, is found to enhance expression of GSH11 in a VDE-dependent manner under conditions of sulfur starvation. These results indicate that GSH11 is regulated by a system sensitive to sulfur starvation (presumably via cysteine depletion) and a more general system involving the nutritional starvation signal mediated by the TOR system. Both systems need to be operational (inhibition of TOR and sulfur starvation) for full expression of GSH11. PMID- 12900423 TI - Extended substrate specificity of rat mast cell protease 5, a rodent alpha chymase with elastase-like primary specificity. AB - Chymases are mast cell serine proteases with chymotrypsin-like primary substrate specificity. Amino acid sequence comparisons of alpha-chymases from different species indicated that certain rodent alpha-chymases have a restricted S1 pocket that could only accommodate small amino acids, i.e. they may, despite being classified as chymases, in fact display elastase-like substrate specificity. To explore this possibility, the alpha-chymase, rat mast cell protease 5 (rMCP-5), was produced as a proenzyme with a His6 purification tag and an enterokinase susceptible peptide replacing the natural propeptide. After removal of the purification tag/enterokinase site by enterokinase digestion, rMCP-5 bound the serine-protease-specific inhibitor diisopropyl fluorophosphate, showing that rMCP 5 was catalytically active. The primary specificity was investigated with chromogenic substrates of the general sequence succinyl-Ala-Ala-Pro-X-p nitroanilide, where the X was Ile, Val, Ala, Phe or Leu. The activity was highest toward substrates with Val or Ala in the P1 position, whereas low activity toward the peptide with a P1 Phe was observed, indicating that the substrate specificity of rMCP-5 indeed is elastase-like. The extended substrate specificity was examined utilizing a phage-displayed random nonapeptide library. The preferred cleavage sequence was resolved as P4-(Gly/Pro/Val), P3-(Leu/Val/Glu), P2 (Leu/Val/Thr), P1-(Val/Ala/Ile), P1'-(Xaa), and P2'-(Glu/Leu/Asp). Hence, the extended substrate specificity is similar to human chymase in most positions except for the P1 position. We conclude that the rat alpha-chymase has converted to elastase-like substrate specificity, perhaps associated with an adoption of new biological targets, separate from those of human alpha-chymase. PMID- 12900424 TI - A novel interaction between perlecan protein core and progranulin: potential effects on tumor growth. AB - In an in vivo search of novel partners for perlecan, a major heparan sulfate proteoglycan of basement membranes and cell surfaces, we identified progranulin, a secreted growth factor, as a strong interacting protein. Unambiguous interaction, first observed with the yeast two-hybrid system, was corroborated by co-immunoprecipitation studies using cell-free transcription/translation and transient cell transfection assays. The interaction of progranulin with perlecan domain V involved the first two laminin- and epidermal growth factor-like repeats. Within progranulin, the subdomains interacting most with perlecan harbored granulins F and B. Kinetics analysis of the interaction using surface plasmon resonance showed a saturable binding of relative low affinity (KD approximately 1 microM). These results were supported by significant expression overlap of these two proteins in a series of ovarian tumor tissue microarrays. Progranulin was present within proliferating blood vessels of ovarian carcinomas and perivascular matrices, with a distribution similar to perlecan. Notably, both progranulin and domain V stimulated the growth of adrenal carcinoma cells. However, when used together in equimolar amounts, the two proteins counteracted each other's activity. Thus, progranulin/perlecan interaction could contribute to a fine regulation of tumor angiogenesis and could ultimately affect cancer growth. PMID- 12900426 TI - Is there any advantage to using an arterial transfer function? PMID- 12900427 TI - Amlodipine modulates THP-1 cell adhesion to vascular endothelium via inhibition of protein kinase C signal transduction. AB - Inflammatory responses play an important role in atherosclerosis. To critically assess the effect of dihydropyridines in inflammatory reactions, we conducted a monocyte-endothelial adhesion assay with monocytic THP-1 cells treated with amlodipine under flow conditions in vitro. THP-1 cells were incubated in the presence of amlodipine (10 micromol/L) for 48 hours and then perfused over activated (interleukin-1beta, 10 U/mL, 4 hours) human umbilical vein endothelial cells. The adhesion of THP-1 cells was significantly reduced after amlodipine treatment (P<0.001); however, flow cytometric analysis reveled that the expression levels of integrins in THP-1 cells were not significantly altered. Furthermore, Western blotting analysis of THP-1 cell lysates revealed that translocation of RhoA from the cytosol to the membrane was significantly diminished after amlodipine treatment. In addition, activation of protein kinase C-alpha and -beta, as well as intracellular calcium influx, induced by phorbol 12 myristate 13-acetate, was diminished after amlodipine treatment. Pretreatment of THP-1 cells with calphostin C, a potent inhibitor of protein kinase C, significantly reduced THP-1 adhesion to vascular endothelium, whereas activation of beta1-integrin was reduced after amlodipine treatment in THP-1 cells, based on the immunoreactivity of an activation-specific antibody for beta1-integrin. Similar inhibitory effects were observed when we used freshly isolated peripheral blood mononuclear cells. These findings suggest a potential role for amlodipine in monocyte-endothelial interactions by modulation of protein kinase C- and RhoA dependent mechanisms, which might account for its vascular protective effects. PMID- 12900428 TI - Recovery of high blood pressure after chronic lesions of the commissural NTS in SHR. AB - Acute electrolytic lesions of the commissural nucleus of the solitary tract (commNTS) reduce blood pressure (BP) in SHR but not in normotensive Wistar-Kyoto and Wistar rats and abolish the pressor response to intravenous injection of potassium cyanide. We investigated the chronic effect of commNTS lesions on mean arterial pressure (MAP), and on baroreceptor and chemoreceptor reflex responses in SHR. The contribution of the sympathetic nervous system and the hormones vasopressin and angiotensin II to maintenance of BP in lesioned SHR was also investigated. MAP fell to normotensive levels the day after lesioning the commNTS but returned to the hypertensive level 9 days later. The reflex tachycardia evoked by sodium nitroprusside remained attenuated for 10 days after commNTS lesions but became enhanced 30 days after commNTS lesions. The pressor component of the chemoreflex elicited by potassium cyanide remained blocked for 30 days after lesions. Vasopressin antagonist or ACE blocker did not change MAP in sham or commNTS-lesioned SHR. Ganglionic blockade with hexamethonium elicited similar reductions in MAP in sham and commNTS-lesioned SHR. Results demonstrated that commNTS lesions in SHR produce a transient fall in BP and a long-lasting inhibition of the pressor response of the chemoreflex. Therefore, the blockade of the pressor response to peripheral chemoreflex activation is not sufficient to chronically reduce MAP in SHR. In the chronic absence of the commNTS, other subnuclei of the NTS or other brain stem nuclei may reorganize to replace the function of commNTS neurons, restoring sympathetic activity and high BP in SHR. PMID- 12900429 TI - Serum heat shock protein 70 levels predict the development of atherosclerosis in subjects with established hypertension. AB - Although heat shock proteins (Hsp's) are present in the sera of healthy individuals and at elevated levels in subjects with early cardiovascular disease, their physiologic role in and value for predicting the development and/or progression of atherosclerosis have not been evaluated. Serum was obtained from 218 subjects with established hypertension (diastolic pressure >95 mm Hg) before their enrollment in the European Lacidipine Study on Atherosclerosis. Hsp60 and Hsp70, and anti-human Hsp60, anti-human Hsp70, and anti-mycobacterial Hsp65 antibody levels were measured by enzyme immunoassay. As an indicator of the presence/progression of atherosclerosis, the means of the maximum intima-media (I M) thicknesses in the far walls of common carotid arteries and bifurcations (CBMmax) were determined by ultrasonography at the time of enrollment and 4 years afterward. Increases in I-M thicknesses at follow-up were less prevalent in subjects having high serum Hsp70 levels (75th percentile) at the time of enrollment (odds ratio, 0.42; 95% confidence interval [CI], 0.22 to 0.8, P=0.008). Although a similar trend was observed for serum Hsp60 levels, this was not statistically significant (odds ratio, 0.6; 95% CI, 0.32 to 1.11, P=0.10). There was no relation between anti-Hsp antibody levels and changes in I-M thicknesses. The relation between Hsp70 levels and changes in I-M thickness was independent of age, atenolol or lacidipine treatment, smoking habits, and blood lipid levels. These findings indicate that circulating Hsp70 levels predict the development of atherosclerosis in subjects with established hypertension, and an intriguing possibility is that Hsp70 protects against or modifies the progression of atherosclerosis in this subject group. PMID- 12900430 TI - Elastase-induced suppression of endothelin-mediated Ca2+ entry mechanisms of vascular contraction. AB - Abdominal aortic aneurysm (AAA) is associated with increased endothelin (ET-1), both systemically and locally in the aorta. Also, elastase activity is increased in human AAA, and elastase perfusion of the aorta induces aneurysm formation in animal models of AAA. However, whether elastase directly affects the ET-1-induced mechanisms of aortic smooth muscle contraction is unclear. Isometric contraction and 45Ca2+ influx were measured in aortic strips isolated from male Sprague Dawley rats and treated with elastase (5 U/mL). To avoid degradation of the extracellular matrix proteins by elastase, experiments were performed in the presence of elastin (10 mg/mL). In normal Krebs solution (2.5 mmol/L Ca2+), ET-1 (10(-7) mol/L) caused contraction of aortic strips that was inhibited by elastase (5 U/mL). The elastase-induced inhibition of ET-1 contraction was slow in onset (4.6+/-0.4 minutes), time-dependent, complete in 34+/-3 minutes, and reversible. In Ca2+-free Krebs solution, caffeine (25 mmol/L) caused a small contraction that was not inhibited by elastase, suggesting that elastase does not inhibit Ca2+ release from the intracellular stores. Membrane depolarization by 96 mmol/L KCl, which stimulates Ca2+ entry from the extracellular space, caused a contraction that was inhibited by elastase in a concentration-dependent, time-dependent, and reversible fashion. The reversible inhibitory effects of elastase, particularly in the presence of elastin, suggest that they are not due to dissolution of the extracellular matrix or smooth muscle contractile proteins. Elastase also inhibited ET-1 and KCl-induced 45Ca2+ influx. Thus, elastase directly inhibits ET 1-induced Ca2+ entry mechanisms of vascular smooth muscle contraction, which may explain the role of elastase and ET-1 during the development of AAA. PMID- 12900431 TI - Hyperuricemia in childhood primary hypertension. AB - Experimental animal models suggest that uric acid might have a pathogenic role in the early development of primary hypertension. We hypothesized that serum uric acid is correlated with blood pressure in children with new-onset, untreated, primary hypertension. We evaluated 125 consecutive children referred to the Baylor Pediatric Renal Program for evaluation of hypertension. None of the subjects had previously been evaluated or treated for hypertension. The children ranged in age from 6 to 18 years (mean, 13.4+/-3.3) and had normal renal function (creatinine clearance >80 mL x min(-1) x 1.73 m(-2)). Sixty-three children had primary hypertension, 40 had secondary hypertension, and 22 had white-coat hypertension. Forty controls with normal blood pressure were recruited from the renal clinic. Uric acid levels were directly correlated with systolic (r=0.80, P=0.0002) and diastolic (r=0.66, P=0.0006) blood pressure in controls and in subjects with primary hypertension and were independent of renal function. Serum uric acid concentrations >5.5 mg/dL were found in 89% of subjects with primary hypertension, in 30% with secondary hypertension, in 0% with white-coat hypertension, and in 0% of controls. We conclude that serum uric acid is directly correlated with blood pressure in untreated children and that a serum uric acid value >5.5 mg/dL in an adolescent being evaluated for hypertension strongly suggests primary hypertension as opposed to white-coat or secondary hypertension. These results are consistent with the hypothesis that uric acid might have a role in the early pathogenesis of primary hypertension. PMID- 12900432 TI - Effects of exogenous heme on renal function: role of heme oxygenase and cyclooxygenase. AB - We examined the effects of heme administration (15 mg/kg IV) on indexes of renal carbon monoxide production and contrasted the renal functional response to heme in anesthetized rats pretreated and not pretreated with stannous mesoporphyrin (40 micromol/kg IV) to inhibit heme oxygenase or sodium meclofenamate (5 mg/kg IV plus infusion at 10 microg/kg per minute) to inhibit cyclooxygenase. In rats without drug pretreatment, heme administration decreased renal vascular resistance and increased renal blood flow, urine volume, and sodium excretion associated with augmented urinary excretion of 6-keto-PGF1alpha and enhanced concentration of carbon monoxide in the renal cortical microdialysate. Pretreatment with stannous mesoporphyrin did not prevent heme from producing renal vasodilation and increasing renal blood flow but abolished the diuretic and natriuretic responses. Conversely, pretreatment with sodium meclofenamate blunted the renal vasodilatory effect of heme but affected neither the diuretic nor the natriuretic effect. We conclude that heme-induced renal vasodilation is a cyclooxygenase-dependent response involving increased synthesis of PGI2, whereas heme-induced diuresis and natriuresis are heme oxygenase-dependent responses involving inhibition of tubular reabsorption of sodium and water through undefined mechanisms. PMID- 12900433 TI - N-domain angiotensin I-converting enzyme with 80 kDa as a possible genetic marker of hypertension. AB - We have previously described angiotensin I-converting enzyme (ACE) forms in urine of normotensive (190 and 65 kDa) and hypertensive patients (90 and 65 kDa, N domain ACEs). Based on the results described above, experimental and genetic models of hypertension were investigated to distinguish hemodynamic and genetic influence on the generation of ACE profile in urine: Wistar-Kyoto and Brown Norway rats (WKY and BN), spontaneously and stroke-prone spontaneously hypertensive rats (SHR and SHR-SP), one kidney/one clip rats (1K1C), deoxycorticosterone acetate (DOCA) salt-treated and untreated rats, and enalapril treated SHR (SHRen). Two peaks with ACE activity were separated from the urine of WKY and BN rats submitted to an AcA-44 column, WK-1/BN-1 (190 kDa), and WK-2/BN-2 (65 kDa), as described for urine of normotensive subjects. The same results were obtained for urine of 1K1C and DOCA salt-treated and untreated rats, analyzed to evaluate the influence of hemodynamic factors in the ACE profile in urine. The urine from SHR, SHR-SP, and SHRen presented 80 (S-1, SP-1, Sen-1) and 65 (S-2, SP 2, Sen-2) kDa ACE forms, differing from the urine profile of normotensive rats, but similar to that described for hypertensive patients. The presence of 80 kDa ACE in urine of SHR, SHR-SP, and SHRen and its absence in urine of experimental hypertensive rats (1K1C and DOCA salt) support the hypothesis that this enzyme could be a possible genetic marker of hypertension. Taken together, our results provide evidence that ACE forms with 90/80 kDa isolated from the urine of hypertensive subjects and genetic hypertensive animals behaves as a possible genetic marker of hypertension and not as a marker of high blood pressure. PMID- 12900434 TI - Pregnant rats with 5/6 nephrectomy have normal volume expansion despite lower renin and kallikrein. AB - To test the hypothesis that normal volume expansion during pregnancy is impaired during chronic renal failure, we evaluated the effects of 5/6 nephrectomy (Nx) in Sprague-Dawley rats. Partial Nx was produced by ligation of 2/3 renal arteries and contralateral Nx. Control rats had a sham operation. Four weeks later, rats were assigned to nonpregnant (n=6/each) or pregnant groups (n=11 to 12/each). At day 21, blood pressure, plasma volume, renal function, hormonal levels, and reproductive outcome were determined. Statistical analysis was performed by ANOVA, and values were expressed as mean+/-SEM. Rats with 5/6 Nx had proteinuria and lower creatinine clearance; pregnancy did not affect these parameters. Blood pressure increased in 5/6 Nx rats and was reduced by pregnancy. Both pregnant groups had lower hematocrit and higher plasma volume values (nonpregnant control, 13.4+/-0.2; nonpregnant 5/6 Nx, 14.4+/-0.2; pregnant control, 19.1+/-0.7, pregnant 5/6 Nx, 19+/-0.9 mL, P<0.001). Pregnancy increased plasma renin activity only in control rats (nonpregnant control, 2.0+/-0.7; nonpregnant 5/6 Nx, 1.6+/ 1.1; pregnant control, 36.1+/-7.9, pregnant 5/6 Nx, 6.1+/-1.8 ng AI/mL per hour, P<0.001). Serum aldosterone levels were unaffected by 5/6 Nx and were higher in pregnant than in nonpregnant rats. Urinary kallikrein activity was reduced by 5/6 Nx and not changed by pregnancy (nonpregnant control, 1499+/-237; nonpregnant 5/6 Nx, 346+/-90; pregnant control, 1595+/-180, pregnant 5/6 Nx, 374+/-95 nmol/16 hours, P<0.001). No significant differences in fetal and placental weights were observed between control and 5/6 Nx rats. Present data indicate that 5/6 Nx pregnant rats were able to normally expand plasma volume despite lower renin and kallikrein levels. PMID- 12900435 TI - Role of COX-2 in the enhanced vasoconstrictor effect of arachidonic acid in the diabetic rat kidney. AB - In the rat isolated perfused kidney, arachidonic acid elicits cyclooxygenase dependent vasoconstriction through activation of PGH2/TxA2 receptors; responses are enhanced in kidneys from diabetic rats. This study examined the roles of cyclooxygenase-1/cyclooxygenase-2 in the enhanced renal vasoconstrictor effect of arachidonic acid in streptozotocin-diabetic rats. Release of 20-HETE was also determined, as this eicosanoid has been reported to elicit cyclooxygenase dependent vasoconstriction. We confirmed that vasoconstrictor responses to arachidonic acid were enhanced in the diabetic rat kidney associated with a 2 fold-greater increase in the release of 6-ketoPGF1alpha, which was used as an index of cyclooxygenase activity. One and three micrograms of arachidonic acid increased perfusion pressure by 85+/-37 and 186+/-6 mm Hg, respectively, in diabetic rat kidneys compared with 3+/-1 and 17+/-8 mm Hg, respectively, in control rat kidneys. Inhibition of both cyclooxygenase isoforms with indomethacin (10 micromol/L) abolished the vasoconstrictor response to arachidonic acid in both diabetic and control rat kidneys, whereas inhibition of cyclooxygenase-2 with nimesulide (5 micromol/L) reduced perfusion pressure responses to 1 and 3 microg arachidonic acid only in the diabetic rat kidney to 15+/-8 and 108+/-26 mm Hg, respectively, consistent with a 3-fold increase in the renal cortical expression of cyclooxygenase-2. 20-HETE release from the diabetic rat kidney was reduced almost 6-fold and was not increased in response to arachidonic acid. These results demonstrate that the renal vasoconstrictor effect of arachidonic acid is solely dependent on cyclooxygenase activity, with no evidence for a contribution from 20-HETE; in the diabetic rat, cyclooxygenase-2 activity contributes to the renal vasoconstrictor effect of arachidonic acid. PMID- 12900436 TI - Salt-sensitive hypertension after exposure to angiotensin is associated with inability to upregulate renal epoxygenases. AB - The current study was designed to determine whether angiotensin II infusion could lead to persistent salt-sensitive hypertension and to examine involvement of renal microvascular epoxygenases in this process. Six groups were studied: rats maintained on a normal salt diet for 4 weeks (NS); rats maintained on a high salt diet for 4 weeks (HS); and all other animals receiving angiotensin II (ANG) infusion and being fed a normal or high salt diet for 2 weeks; then the angiotensin II infusion was stopped and diets were either maintained or switched (ANG/NS-NS, ANG/NS-HS, ANG/HS-HS, ANG/HS-NS). Angiotensin II infusion resulted in a rise in blood pressure and an increase in urinary albumin excretion over the 2 week period. After angiotensin II withdrawal, blood pressure returned to normal in animals receiving a normal salt diet from weeks 2 to 4 (ANG/NS-NS and ANG/HS NS groups). In contrast, blood pressure remained elevated in the group maintained on a high salt diet throughout the entire 4-week period (ANG/HS-HS group). Renal microvascular CYP2C11 and CYP2C23 protein levels were decreased by 50% to 60% in the ANG/HS-HS group compared with the NS group. Likewise, renal microvascular CYP2J protein was significantly decreased in the ANG/HS-HS group versus the NS group. Renal microvascular CYP2C11 and CYP2C23 mRNA levels were reduced in the ANG/HS-HS group compared with both the NS and HS groups. These results support the hypothesis that angiotensin II infusion induces persistent salt-sensitive hypertension after withdrawal of angiotensin II that may be due to downregulation of CYP2C and CYP2J epoxygenases in renal microvessels. PMID- 12900437 TI - Beta2 adrenoceptor functional gene variants, obesity, and blood pressure level interactions in the general population. AB - We investigated the association of beta2 adrenoceptor functional gene variants (Arg16Gly, Gln27Glu, and Thr164Ile polymorphisms), obesity phenotypes, and blood pressure levels in a large, ethnically mixed urban population. The individuals (n=1576) were randomly selected for a cross-sectional study of cardiovascular risk factors in Vitoria, Brazil. Statistically significant associations among systolic blood pressure and the Arg16Gly and Thr164Ile variants were identified in univariate analysis. The Gly16/Gly16 genotype was still associated with systolic blood pressure (SBP) in multivariate analysis adjusting for age, gender, ethnicity, total cholesterol, diabetes, and body mass index (BMI) (P=0.01). The Arg16 allele was the only genotypic variable associated with BMI, and, in a dominant model, it remained associated with an increased BMI even after adjustment for age, gender, ethnicity, triglycerides, HDL cholesterol, LDL cholesterol, diabetes, and hypertension status (P=0.02). Although the different polymorphisms did not interact in the determination of SBP, a significant interaction with BMI (P=0.02), not through linkage disequilibrium, was identified between the Gln27Glu and the Thr164Ile variants. Furthermore, a significant interaction among the Arg16Gly polymorphism and BMI (P=0.036) and waist-hip ratio (P=0.003) in determining SBP was disclosed by ANOVA factorial modeling, with SBP used as the dependent variable. An interaction between the Thr164Ile polymorphism and waist-hip ratio was also identified (P=0.018). Finally, multiple logistic regression models showed a 1.48-fold increase in the risk of hypertension in individuals harboring the Gly16/Gly16 genotype and a 1.31-fold (P=0.01) and a 1.49-fold (P=0.003) increased risk of obesity in individuals harboring the Gln27/Gln27 genotype or the presence of the Arg16 allele, respectively. Taken together, these data provide evidence for a strong but complex relation between beta-adrenoceptor gene variants, hypertension, and obesity. PMID- 12900438 TI - Perturbation of D1 dopamine and AT1 receptor interaction in spontaneously hypertensive rats. AB - The dopaminergic and renin-angiotensin systems interact to regulate blood pressure. Because this interaction may be perturbed in genetic hypertension, we studied D1 dopamine and AT1 angiotensin receptors in immortalized renal proximal tubule (RPT) and A10 aortic vascular smooth muscle cells. In normotensive Wistar Kyoto (WKY) rats, the D1-like agonist fenoldopam increased D1 receptors but decreased AT1 receptors. These effects were blocked by the D1-like antagonist SCH 23390 (10(-7) mol/L per 24 hours). In spontaneously hypertensive rat (SHR) RPT cells, fenoldopam also decreased AT1 receptors but no longer stimulated D1 receptor expression. Basal levels of AT1/D1 receptor coimmunoprecipitation were greater in WKY RPT cells (29+/-2 density units, DU) than in SHR RPT cells (21+/-2 DU, n=7 per group, P<0.05). The coimmunoprecipitation of D1 and AT1 receptors was increased by fenoldopam (10(-7) mol/L per 24 hours) in WKY RPT cells but decreased in SHR RPT cells. The effects of fenoldopam in RPT cells from WKY rats were similar in aortic vascular smooth muscle cells from normotensive BD IX rats, that is, fenoldopam decreased AT1 receptors and increased D1 receptors. Our studies show differential regulation of the expression of D1 and AT1 receptors in RPT cells from WKY and SHR. This regulation and D1/AT1 receptor interaction are different in RPT cells of WKY and SHR. An altered interaction of D1 and AT1 receptors may play a role in the impaired sodium excretion and enhanced vasoconstriction in hypertension. PMID- 12900439 TI - Sympathoexcitation by PVN-injected bicuculline requires activation of excitatory amino acid receptors. AB - Acute blockade of gamma-aminobutyric acid (GABA)-A receptors in the hypothalamic paraventricular nucleus (PVN) increases mean arterial pressure (MAP), heart rate (HR), and sympathetic nerve activity (SNA). However, the underlying neural mechanisms have not been fully determined. We tested the hypothesis that responses to GABA-A receptor blockade in the PVN require activation of local ionotropic excitatory amino acid (EAA) receptors. MAP, HR, and renal SNA responses to unilateral PVN microinjection of bicuculline methobromide (BIC, 0.1 nmol) were recorded before and after ipsilateral PVN injection of either vehicle (saline), the nonselective ionotropic EAA receptor antagonist kynurenate (KYN), the NMDA receptor antagonist D(-)-2-amino-5-phosphonopentanoic acid (AP5), or the non-NMDA receptor antagonist 2,3-dioxo-6-nitro-1,2,3,4 tetrahydrobenzo[f]quinoxaline-7-sulfonamide disodium (NBQX). Responses to PVN injected BIC were unaltered by vehicle injection. In contrast, injection of KYN (7.2 nmol; n=4) nearly abolished ABP and renal SNA responses to BIC (P<0.01) and significantly attenuated (P<0.05) HR responses as well. Similarly, graded doses of AP5 (0.6, 3, and 6 nmol) and NBQX (0.26, 1.3, and 2.6 nmol) reduced responses to PVN-injected BIC in a dose-related manner, with the 3 nmol (n=7) and 1.3 nmol (n=6) doses producing maximal effects (P<0.05). KYN, AP5, and NBQX did not affect baseline parameters. Effects of a cocktail containing AP5 (3 nmol) and NBQX (1.3 nmol) were greater (P<0.01) than either antagonist alone and were not statistically different from KYN. These data indicate that cardiovascular and renal sympathetic responses to acute GABA-A receptor blockade in the PVN require local actions of EAAs at both NMDA and non-NMDA receptors. PMID- 12900440 TI - Embryo morphogenesis: getting down to cells and molecules. PMID- 12900441 TI - Consequences of the depletion of zygotic and embryonic enhancer of zeste 2 during preimplantation mouse development. AB - Enhancer of zeste 2 (Ezh2), a SET domain-containing protein, is crucial for development in many model organisms, including early mouse development. In mice, Ezh2 is detected as a maternally inherited protein in the oocyte but its function at the onset of development is unknown. We have used a conditional allele of Ezh2 to deplete the oocyte of this maternal inheritance. We show that the loss of maternal Ezh2 has a long-term effect causing severe growth retardation of neonates despite 'rescue' through embryonic transcription from the paternal allele. This phenotypic effect on growth could be attributed to the asymmetric localisation of the Ezh2/Eed complex and the associated histone methylation pattern to the maternal genome, which is disrupted in Ezh2 mutant zygotes. During subsequent development, we detect distinct histone methylation patterns in the trophectoderm and the pluripotent epiblast. In the latter where Oct4 expression continues from the zygote onwards, the Ezh2/Eed complex apparently establishes a unique epigenetic state and plasticity, which probably explains why loss of Ezh2 is early embryonic lethal and obligatory for the derivation of pluripotent embryonic stem cells. By contrast, in the differentiating trophectoderm cells where Oct4 expression is progressively downregulated Ezh2/Eed complex is recruited transiently to one X chromosome in female embryos at the onset of X inactivation. This accumulation and the associated histone methylation are also lost in Ezh2 mutants, suggesting a role in X inactivation. Thus, Ezh2 has significant and diverse roles during early development, as well as during the establishment of the first differentiated cells, the trophectoderm, and of the pluripotent epiblast cells. PMID- 12900442 TI - The Arabidopsis ACR4 gene plays a role in cell layer organisation during ovule integument and sepal margin development. AB - The mechanisms regulating cell layer organisation in developing plant organs are fundamental to plant growth, but remain largely uninvestigated. We have studied the receptor kinase-encoding ARABIDOPSIS CRINKLY4 gene and shown that its expression is restricted to the L1 cell layer of most meristems and organ primordia, including those of the ovule integuments. Insertion mutations show that ARABIDOPSIS CRINKLY4 is required for regulation of cellular organisation during the development of sepal margins and ovule integument outgrowth. We show that ARABIDOPSIS CRINKLY4 encodes a functional kinase that, in ovules and possibly other tissues, is abundant in anticlinal and the inner periclinal plasma membrane of 'outside' cells. We propose that ARABIDOPSIS CRINKLY4 may be involved in maintaining L1 cell layer integrity by receiving and transmitting signals from neighbouring L1 cells and/or from underlying cell layers. PMID- 12900443 TI - Feedback loops comprising Dll1, Dll3 and Mesp2, and differential involvement of Psen1 are essential for rostrocaudal patterning of somites. AB - Elaborate metamerism in vertebrate somitogenesis is based on segmental gene expression in the anterior presomitic mesoderm (PSM). Notch signal pathways with Notch ligands Dll1 and Dll3, and the transcription factor Mesp2 are implicated in the rostrocaudal patterning of the somite. We have previously shown that changes in the Mesp2 expression domain from a presumptive one somite into a rostral half somite results in differential activation of two types of Notch pathways, dependent or independent of presenilin 1 (Psen1), which is a Notch signal mediator. To further refine our hypothesis, we have analyzed genetic interactions between Dll1, Dll3, Mesp2 and Psen1, and elucidated the roles of Dll1- and Dll3 Notch pathways, with or without Psen1, in rostrocaudal patterning. Dll1 and Dll3 are co-expressed in the PSM and so far are considered to have partially redundant functions. We find in this study that positive and negative feedback loops comprising Dll1 and Mesp2 appear to be crucial for this patterning, and Dll3 may be required for the coordination of the Dll1-Mesp2 loop. Additionally, our epistatic analysis revealed that Mesp2 affects rostrocaudal properties more directly than Dll1 or Dll3. Finally, we find that Psen1 is involved differently in the regulation of rostral and caudal genes. Psen1 is required for Dll1-Notch signaling for activation of Dll1, while the Psen1-independent Dll3-Notch pathway may counteract the Psen1-dependent Dll1-Notch pathway. These observations suggest that Dll1 and Dll3 may have non-redundant, even counteracting functions. We conclude from our analyses that Mesp2 functions as a central mediator of such Notch pathways and regulates the gene expression required for rostrocaudal patterning of somites. PMID- 12900444 TI - Anterior and posterior waves of cyclic her1 gene expression are differentially regulated in the presomitic mesoderm of zebrafish. AB - Somite formation in vertebrates depends on a molecular oscillator in the presomitic mesoderm (PSM). In order to get a better insight into how oscillatory expression is achieved in the zebrafish Danio rerio, we have analysed the regulation of her1 and her7, two bHLH genes that are co-expressed in the PSM. Using specific morpholino oligonucleotide mediated inhibition and intron probe in situ hybridisation, we find that her7 is required for initiating the expression in the posterior PSM, while her1 is required to propagate the cyclic expression in the intermediate and anterior PSM. Reporter gene constructs with the her1 upstream sequence driving green fluorescent protein (GFP) expression show that separable regulatory regions can be identified that mediate expression in the posterior versus intermediate and anterior PSM. Our results indicate that the cyclic expression is generated at the transcriptional level and that the resulting mRNAs have a very short half-life. A specific degradation signal for her1 mRNA must be located in the 5'-UTR, as this region also destabilises the GFP mRNA such that it mimics the dynamic pattern of the endogenous her1 mRNA. In contrast to the mRNA, GFP protein is stable and we find that all somitic cells express the protein, proving that her1 mRNA is transiently expressed in all cells of the PSM. PMID- 12900445 TI - The chemokine SDF1/CXCL12 and its receptor CXCR4 regulate mouse germ cell migration and survival. AB - In mouse embryos, germ cells arise during gastrulation and migrate to the early gonad. First, they emerge from the primitive streak into the region of the endoderm that forms the hindgut. Later in development, a second phase of migration takes place in which they migrate out of the gut to the genital ridges. There, they co-assemble with somatic cells to form the gonad. In vitro studies in the mouse, and genetic studies in other organisms, suggest that at least part of this process is in response to secreted signals from other tissues. Recent genetic evidence in zebrafish has shown that the interaction between stromal cell derived factor 1 (SDF1) and its G-protein-coupled receptor CXCR4, already known to control many types of normal and pathological cell migrations, is also required for the normal migration of primordial germ cells. We show that in the mouse, germ cell migration and survival requires the SDF1/CXCR4 interaction. First, migrating germ cells express CXCR4, whilst the body wall mesenchyme and genital ridges express the ligand SDF1. Second, the addition of exogenous SDF1 to living embryo cultures causes aberrant germ cell migration from the gut. Third, germ cells in embryos carrying targeted mutations in CXCR4 do not colonize the gonad normally. However, at earlier stages in the hindgut, germ cells are unaffected in CXCR4(-/-) embryos. Germ cell counts at different stages suggest that SDF1/CXCR4 interaction also mediates germ cell survival. These results show that the SDF1/CXCR4 interaction is specifically required for the colonization of the gonads by primordial germ cells, but not for earlier stages in germ cell migration. This demonstrates a high degree of evolutionary conservation of part of the mechanism, but also an area of evolutionary divergence. PMID- 12900446 TI - GP130, the shared receptor for the LIF/IL6 cytokine family in the mouse, is not required for early germ cell differentiation, but is required cell-autonomously in oocytes for ovulation. AB - GP130 is the shared receptor for members of the IL6 family of cytokines. Members of this family have been shown to enhance the survival of migratory (E10.5) or postmigratory (E12.5) murine primordial germ cells (PGCs) in culture; however, it is uncertain what role these cytokines play during PGC development in vivo. We have examined PGC numbers in E13.5 GP130-deficient mouse embryos and found that males exhibited a slight decrease in PGC numbers; females were normal. Also, we used the Cre-loxP system to inactive GP130 specifically in germ cells and found that this resulted in a fertility defect in females. These animals were found to have a slight reduction in the number of primary follicles and a major defect in ovulation. This data suggests that GP130 is required in female germ cells for their normal function, but is dispensable in male germ cells. PMID- 12900447 TI - Wise, a context-dependent activator and inhibitor of Wnt signalling. AB - We have isolated a novel secreted molecule, Wise, by a functional screen for activities that alter the anteroposterior character of neuralised Xenopus animal caps. Wise encodes a secreted protein capable of inducing posterior neural markers at a distance. Phenotypes arising from ectopic expression or depletion of Wise resemble those obtained when Wnt signalling is altered. In animal cap assays, posterior neural markers can be induced by Wnt family members, and induction of these markers by Wise requires components of the canonical Wnt pathway. This indicates that in this context Wise activates the Wnt signalling cascade by mimicking some of the effects of Wnt ligands. Activation of the pathway was further confirmed by nuclear accumulation of beta-catenin driven by Wise. By contrast, in an assay for secondary axis induction, extracellularly Wise antagonises the axis-inducing ability of Wnt8. Thus, Wise can activate or inhibit Wnt signalling in a context-dependent manner. The Wise protein physically interacts with the Wnt co-receptor, lipoprotein receptor-related protein 6 (LRP6), and is able to compete with Wnt8 for binding to LRP6. These activities of Wise provide a new mechanism for integrating inputs through the Wnt coreceptor complex to modulate the balance of Wnt signalling. PMID- 12900448 TI - Tracing of her5 progeny in zebrafish transgenics reveals the dynamics of midbrain hindbrain neurogenesis and maintenance. AB - The midbrain-hindbrain domain (MH) of the vertebrate embryonic neural tube develops in response to the isthmic organizer (IsO), located at the midbrain hindbrain boundary (MHB). MH derivatives are largely missing in mutants affected in IsO activity; however, the potentialities and fate of MH precursors in these conditions have not been directly determined. To follow the dynamics of MH maintenance in vivo, we used artificial chromosome transgenesis in zebrafish to construct lines where egfp transcription is driven by the complete set of regulatory elements of her5, the first known gene expressed in the MH area. In these lines, egfp transcription faithfully recapitulates her5 expression from its induction phase onwards. Using the stability of GFP protein as lineage tracer, we first demonstrate that her5 expression at gastrulation is a selective marker of MH precursor fate. By comparing GFP protein and her5 transcription, we further reveal the spatiotemporal dynamics of her5 expression that conditions neurogenesis progression towards the MHB over time. Finally, we trace the molecular identity of GFP-positive cells in the acerebellar (ace) and no-isthmus (noi) mutant backgrounds to analyze directly fgf8 and pax2.1 mutant gene activities for their ultimate effect on cell fate. We demonstrate that most MH precursors are maintained in both mutants but express abnormal identities, in a manner that strikingly differs between the ace and noi contexts. Our observations directly support a role for Fgf8 in protecting anterior tectal and metencephalic precursors from acquiring anterior identities, while Pax2.1 controls the choice of MH identity as a whole. Together, our results suggest a model where an ordered MH pro-domain is identified at gastrulation, and where cell identity choices within this domain are subsequently differentially controlled by Fgf8 and Pax2.1 functions. PMID- 12900449 TI - Coherent development of dermomyotome and dermis from the entire mediolateral extent of the dorsal somite. AB - We have previously shown that overall growth of the myotome in the mediolateral direction occurs in a coherent and uniform pattern. We asked whether development of the dermomyotome and resultant dermis follow a similar pattern or are, alternatively, controlled by restricted pools of stem cells driving directional growth. To this end, we studied cellular events that govern dermomyotome development and the regional origin of dermis. Measurements of cell proliferation, nuclear density and cellular rearrangements revealed that the developing dermomyotome can be subdivided in the transverse plane into three distinct and dynamic regions: medial, central and lateral, rather than simply into epaxial and hypaxial domains. To understand how these temporally and spatially restricted changes affect overall dermomyotome growth, lineage tracing with CM-DiI was performed. A proportional pattern of growth was measured along the entire epithelium, suggesting that mediolateral growth of the dermomyotome is coherent. Hence, they contrast with a stem cell view suggesting focal and inversely oriented sources of growth restricted to the medial and lateral edges. Consistent with this uniform mediolateral growth, lineage tracing experiments showed that the dermomyotome-derived dermis originates from progenitors that reside along the medial as well as the lateral halves of somites, and whose contribution to dermis is regionally restricted. Taken together, our results support the view that all derivatives of the dorsal somite (dermomyotome, myotome and dermis) keep a direct topographical relationship with their epithelial ascendants. PMID- 12900450 TI - Unique and combinatorial functions of Fgf3 and Fgf8 during zebrafish forebrain development. AB - Complex spatiotemporal expression patterns of fgf3 and fgf8 within the developing zebrafish forebrain suggest their involvement in its regionalisation and early development. These factors have unique and combinatorial roles during development of more posterior brain regions, and here we report similar findings for the developing forebrain. We show that Fgf8 and Fgf3 regulate different aspects of telencephalic development, and that Fgf3 alone is required for the expression of several telencephalic markers. Within the diencephalon, Fgf3 and Fgf8 act synergistically to pattern the ventral thalamus, and are implicated in the regulation of optic stalk formation, whereas loss of Fgf3 alone results in defects in ZLI development. Forebrain commissure formation was abnormal in the absence of either Fgf3 or Fgf8; however, most severe defects were observed in the absence of both. Defects were observed in patterning of both the midline territory, within which the commissures normally form, and neuronal populations, whose axons comprise the commissures. Analysis of embryos treated with an FGFR inhibitor suggests that continuous FGF signalling is required from gastrulation stages for normal forebrain patterning, and identifies additional requirements for FGFR activity. PMID- 12900451 TI - Developmental regulation and significance of KNOX protein trafficking in Arabidopsis. AB - Intercellular communication delivers critical information for position-dependent specification of cell fate. In plants, a novel mechanism for cell-to-cell communication involves the intercellular trafficking of regulatory proteins and mRNAs. The maize KNOTTED1 (KN1) gene acts non cell-autonomously in the maize leaf, and KN1 was the first plant protein shown to traffic cell-to-cell, presumably through plasmodesmata. We have compared the intercellular trafficking of green fluorescent protein (GFP) fusions of KN1 and Arabidopsis KN1-related homeobox proteins to that of the viral movement protein from turnip vein clearing tobamovirus. We show that there is specific developmental regulation of GFP approximately KN1 trafficking. GFP -- KN1 was able to traffic from the inner layers of the leaf to the epidermis, but not in the opposite direction, from epidermis to mesophyll. However, GFP or the GFP -- movement protein fusion moved readily out of the epidermis. GFP -- KN1 was however able to traffic out of the epidermal (L1) layer in the shoot apical meristem, indicating that KN1 movement out of the L1 was developmentally regulated. GFP -- KNAT1/BREVIPEDICELLUS and GFP -- SHOOTMERISTEMLESS fusions could also traffic from the L1 to the L2/L3 layers of the meristem. In a test for the functional significance of trafficking, we showed that L1-specific expression of KN1 or of KNAT1 was able to partially complement the strong shootmeristemless-11 (stm-11) mutant. However, a cell autonomous GUS fusion to KN1 showed neither trafficking ability nor complementation of stm-11 when expressed in the L1. These results suggest that the activity of KN1 and related homeobox proteins is maintained following intercellular trafficking, and that trafficking may be required for their normal developmental function. PMID- 12900452 TI - Interaction of Par-6 and Crumbs complexes is essential for photoreceptor morphogenesis in Drosophila. AB - Apicobasal cell polarity is crucial for morphogenesis of photoreceptor rhabdomeres and adherens junctions (AJs) in the Drosophila eye. Crumbs (Crb) is specifically localized to the apical membrane of photoreceptors, providing a positional cue for the organization of rhabdomeres and AJs. We show that the Crb complex consisting of Crb, Stardust (Sdt) and Discs-lost (Dlt) colocalizes with another protein complex containing Par-6 and atypical protein kinase C (aPKC) in the rhabdomere stalk of photoreceptors. Loss of each component of the Crb complex causes age-dependent mislocalization of Par-6 complex proteins, and ectopic expression of Crb intracellular domain is sufficient to recruit the Par-6 complex. We also show that the absence of Par-6 complex proteins results in severe mislocalization and loss of Crb complex. We further demonstrate that Dlt directly binds to Par-6, providing a molecular basis for the mutual dependence of the two complexes. These results suggest that the interaction of Crb and Par-6 complexes is required for the organization and maintenance of apical membranes and AJs of photoreceptors. PMID- 12900453 TI - The expression and function of the achaete-scute genes in Tribolium castaneum reveals conservation and variation in neural pattern formation and cell fate specification. AB - The study of achaete-scute (ac/sc) genes has recently become a paradigm to understand the evolution and development of the arthropod nervous system. We describe the identification and characterization of the ac/sc genes in the coleopteran insect species Tribolium castaneum. We have identified two Tribolium ac/sc genes - achaete-scute homolog (Tc-ASH) a proneural gene and asense (Tc-ase) a neural precursor gene that reside in a gene complex. Focusing on the embryonic central nervous system we find that Tc-ASH is expressed in all neural precursors and the proneural clusters from which they segregate. Through RNAi and misexpression studies we show that Tc-ASH is necessary for neural precursor formation in Tribolium and sufficient for neural precursor formation in Drosophila. Comparison of the function of the Drosophila and Tribolium proneural ac/sc genes suggests that in the Drosophila lineage these genes have maintained their ancestral function in neural precursor formation and have acquired a new role in the fate specification of individual neural precursors. Furthermore, we find that Tc-ase is expressed in all neural precursors suggesting an important and conserved role for asense genes in insect nervous system development. Our analysis of the Tribolium ac/sc genes indicates significant plasticity in gene number, expression and function, and implicates these modifications in the evolution of arthropod neural development. PMID- 12900454 TI - Mutation of the photoreceptor specific homeodomain gene Pph13 results in defects in phototransduction and rhabdomere morphogenesis. AB - The expression and organization of the phototransduction signaling proteins into a specialized light-sensing organelle, the rhabdomere, is required for photoreceptor cells to detect light. We report the characterization of the mutant Pph13(hazy). Pph13 is a homeodomain transcription factor expressed only in photoreceptor cells. Pph13 expression correlates with the differentiation and not specification of photoreceptor cells. In agreement with its expression profile, we find Pph13 is required for both rhabdomere morphogenesis and for the proper detection of light. In addition, we demonstrate that Pph13 exerts its effect by the regulation of photoreceptor specific gene expression. PMID- 12900455 TI - CD41 expression defines the onset of primitive and definitive hematopoiesis in the murine embryo. AB - The platelet glycoprotein IIb (alpha(IIb); CD41) constitutes the alpha subunit of a highly expressed platelet surface integrin protein. We demonstrate that CD41 serves as the earliest marker of primitive erythroid progenitor cells in the embryonic day 7 (E7.0) yolk sac and high-level expression identifies essentially all E8.25 yolk sac definitive hematopoietic progenitors. Some definitive hematopoietic progenitor cells in the fetal liver and bone marrow also express CD41. Hematopoietic stem cell competitive repopulating ability is present in CD41(dim) and CD41(lo/-) cells isolated from bone marrow and fetal liver cells, however, activity is enriched in the CD41(lo/-) cells. CD41(bright) yolk sac definitive progenitor cells co-express CD61 and bind fibrinogen, demonstrating receptor function. Thus, CD41 expression marks the onset of primitive and definitive hematopoiesis in the murine embryo and persists as a marker of some stem and progenitor cell populations in the fetal liver and adult marrow, suggesting novel roles for this integrin. PMID- 12900456 TI - Reduced leaf complexity in tomato wiry mutants suggests a role for PHAN and KNOX genes in generating compound leaves. AB - Recent work on species with simple leaves suggests that the juxtaposition of abaxial (lower) and adaxial (upper) cell fates (dorsiventrality) in leaf primordia is necessary for lamina outgrowth. However, how leaf dorsiventral symmetry affects leaflet formation in species with compound leaves is largely unknown. In four non-allelic dorsiventrality-defective mutants in tomato, wiry, wiry3, wiry4 and wiry6, partial or complete loss of ab-adaxiality was observed in leaves as well as in lateral organs in the flower, and the number of leaflets in leaves was reduced significantly. Morphological analyses and expression patterns of molecular markers for ab-adaxiality [LePHANTASTICA (LePHAN) and LeYABBY B (LeYAB B)] indicated that ab-adaxial cell fates were altered in mutant leaves. Reduction in expression of both LeT6 (a tomato KNOX gene) and LePHAN during post primordial leaf development was correlated with a reduction in leaflet formation in the wiry mutants. LePHAN expression in LeT6 overexpression mutants suggests that LeT6 is a negative regulator of LePHAN. KNOX expression is known to be correlated with leaflet formation and we show that LeT6 requires LePHAN activity to form leaflets. These phenotypes and gene expression patterns suggest that the abaxial and adaxial domains of leaf primordia are important for leaflet primordia formation, and thus also important for compound leaf development. Furthermore, the regulatory relationship between LePHAN and KNOX genes is different from that proposed for simple-leafed species. We propose that this change in the regulatory relationship between KNOX genes and LePHAN plays a role in compound leaf development and is an important feature that distinguishes simple leaves from compound leaves. PMID- 12900457 TI - Drosophila E-cadherin is essential for proper germ cell-soma interaction during gonad morphogenesis. AB - In most animal species, germ cells require intimate contact with specialized somatic cells in the gonad for their proper development. We have analyzed the establishment of germ cell-soma interaction during embryonic gonad formation in Drosophila melanogaster, and find that somatic cells undergo dramatic changes in cell shape and individually ensheath germ cells as the gonad coalesces. Germ cell ensheathment is independent of other aspects of gonad formation, indicating that separate morphogenic processes are at work during gonadogenesis. The cell-cell adhesion molecule Drosophila E-cadherin is essential both for germ cell ensheathment and gonad compaction, and is upregulated in the somatic gonad at the time of gonad formation. Our data indicate that differential cell adhesion contributes to cell sorting and the formation of proper gonad architecture. In addition, we find that Fear of Intimacy, a novel transmembrane protein, is also required for both germ cell ensheathment and gonad compaction. E-cadherin expression in the gonad is dramatically decreased in fear of intimacy mutants, indicating that Fear of Intimacy may be a regulator of E-cadherin expression or function. PMID- 12900458 TI - Kette regulates actin dynamics and genetically interacts with Wave and Wasp. AB - During development of the Drosophila nervous system, kette is required for axonal growth and pathfinding. It encodes a highly conserved homolog of the Nck associated protein 1 (NAP1) that genetically interacts with the Drosophila homolog of Nck, dock. We show that in vivo as well as in tissue culture models most of the Kette protein is found in the cytoplasm where it colocalizes with F actin to which it can bind via its N-terminal domain. Some Kette protein is localized at the membrane and accumulates at focal contact sites. Loss of Kette protein results in the accumulation of cytosolic F-actin. The kette mutant phenotype can be suppressed by reducing the wave gene dose, demonstrating that kette antagonizes wave function. Overexpression of the wild-type Kette protein does not interfere with normal development, whereas expression of an activated, membrane-tethered Kette protein induces the formation of large F-actin bundles in both, tissue culture cells and in vivo. This gain-of-function phenotype is independent of wave but can be suppressed by reducing the wasp gene dose, indicating that Kette is able to regulate Wasp, to which it is linked via the Abelson interactor (Abi). Our data suggest a model where Kette fulfils a novel role in regulating F-actin organization by antagonizing Wave and activating Wasp dependent actin polymerization. PMID- 12900460 TI - Role of Islet1 in the patterning of murine dentition. AB - It is believed that mouse dentition is determined by a prepatterning of the oral epithelium into molar (proximal) and incisor (distal) regions. The LIM homeodomain protein Islet1 (ISL1) is involved in the regulation of differentiation of many cell types and organs. During odontogenesis, we find Islet1 to be exclusively expressed in epithelial cells of the developing incisors but not during molar development. Early expression of Islet1 in presumptive incisor epithelium is coincident with expression of Bmp4, which acts to induce Msx1 expression in the underlying mesenchyme. To define the role of ISL1 in the acquisition of incisor shape, we have analysed regulation of Islet1 expression in mandibular explants. Local application of bone morphogenetic protein 4 (BMP4) in the epithelium of molar territories either by bead implantation or by electroporation stimulated Islet1 expression. Inhibition of BMP signalling with Noggin resulted in a loss of Islet1 expression. Inhibition of Islet1 in distal epithelium resulted in a loss of Bmp4 expression and a corresponding loss of Msx1 expression, indicating that a positive regulatory loop exists between ISL1 and BMP4 in distal epithelium. Ectopic expression of Islet1 in proximal epithelium produces a loss of Barx1 expression in the mesenchyme and resulted in inhibition of molar tooth development. Using epithelial/mesenchymal recombinations we show that at E10.5 Islet1 expression is independent of the underlying mesenchyme whereas at E12.5 when tooth shape specification has passed to the mesenchyme, Islet1 expression requires distal (presumptive incisor) mesenchyme. Islet1 thus plays an important role in regulating distal gene expression during jaw and tooth development. PMID- 12900459 TI - A functional overlap of plasminogen and MMPs regulates vascularization during placental development. AB - Both plasminogen activators and matrix metalloproteinases (MMPs) have been implicated in a variety of developmental processes in the mouse during embryo implantation and placentation. We show here that pharmacological treatment of plasminogen-deficient mice with the broad spectrum MMP inhibitor galardin leads to a high rate of embryonic lethality. Implantation sites from plasminogen deficient galardin-treated mice at 7.5 days post coitus (dpc) showed delay in both decidualization and invasion of maternal vessels into the decidua. At 8.5 dpc, half of the embryos were runted and still at the developmental stage of a 7.5 dpc embryo. Most embryos that escaped these initial defects eventually died, probably from defective vascularization and development of the labyrinth layer of the placenta, although a direct role on embryo development cannot be ruled out. These results demonstrate that the combination of MMPs and plasminogen is essential for the proper development of the placenta. Plasminogen deficiency alone and galardin treatment alone had much less effect and there was a pronounced synergism on both placental vascularization and embryonic lethality, indicating a functional overlap between plasminogen and MMPs. PMID- 12900462 TI - The Pax-homeobox gene eyegone is involved in the subdivision of the thorax of Drosophila. AB - The eyegone (eyg) gene is known to be involved in the development of the eye structures of Drosophila. We show that eyg and its related gene, twin of eyegone (toe), are also expressed in part of the anterior compartment of the adult mesothorax (notum). We report experiments concerning the role of these genes in the notum. In the absence of eyg function the anterior-central region does not develop, whereas ectopic activity of either eyg or toe induces the formation of the anterior-central pattern in the posterior or lateral region of the notum. These results demonstrate that eyg and toe play a role in the genetic subdivision of the notum, although the experiments indicate that eyg exerts the principal function. However, by itself the Eyg product cannot induce the formation of notum patterns; its thoracic function requires co-expression with the Iroquois (Iro) genes. We show that the restriction of eyg activity to the anterior-central region of the wing disc is achieved by the antagonistic regulatory activities of the Iro and pnr genes, which promote eyg expression, and those of the Hh and Dpp pathways, which act as repressors. We argue that eyg is a subordinate gene of the Iro genes, and that pnr mediates their thoracic patterning function. The activity of eyg gives rise to a new notum subdivision that acts upon the pre-extant one generated by the Iro genes and pnr. As a result the notum becomes subdivided into four distinct genetic domains. PMID- 12900461 TI - A Twist-like bHLH gene is a downstream factor of an endogenous FGF and determines mesenchymal fate in the ascidian embryos. AB - Ascidian larvae develop mesenchyme cells in their trunk. A fibroblast growth factor (FGF9/16/20) is essential and sufficient for induction of the mesenchyme in Ciona savignyi. We have identified two basic helix-loop-helix (bHLH) genes named Twist-like1 and Twist-like2 as downstream factors of this FGF. These two genes are phylogenetically closely related to each other, and were expressed specifically in the mesenchymal cells after the 110-cell stage. Gene-knockdown experiments using a specific morpholino oligonucleotide demonstrated that Twist like1 plays an essential role in determination of the mesenchyme and that Twist like2 is a downstream factor of Twist-like1. In addition, both overexpression and misexpression of Twist-like1 converts non-mesenchymal cells to mesenchymal cells. We also demonstrate that the upstream regulatory mechanisms of Twist-like1 are different between B-line mesenchymal cells and the A-line mesenchymal cells called 'trunk lateral cells'. FGF9/16/20 is required for the expression of Twist like1 in B-line mesenchymal precursor cells, whereas FGF, FoxD and another novel bHLH factor called NoTrlc are required for Twist-like1 to be expressed in the A line mesenchymal precursor cells. Therefore, two different but partially overlapping mechanisms are required for the expression of Twist-like1 in the mesenchymal precursors, which triggers the differentiation of the mesenchyme in Ciona embryos. PMID- 12900463 TI - Mechanism of inhibition of the Drosophila and mammalian EGF receptors by the transmembrane protein Kekkon 1. AB - The transmembrane protein Kekkon 1 (Kek1) has previously been shown to act in a negative feedback loop to downregulate the Drosophila Epidermal Growth Factor Receptor (DER) during oogenesis. We show that this protein plays a similar role in other DER-mediated developmental processes. Structure-function analysis reveals that the extracellular Leucine-Rich Repeat (LRR) domains of Kek1 are critical for its function through direct association with DER, whereas its cytoplasmic domain is required for apical subcellular localization. In addition, the use of chimeric proteins between Kek1 extracellular and transmembrane domains fused to DER intracellular domain indicates that Kek1 forms an heterodimer with DER in vivo. To characterize more precisely the mechanism underlying the Kek1/DER interaction, we used mammalian ErbB/EGFR cell-based assays. We show that Kek1 is capable of physically interacting with each of the known members of the mammalian ErbB receptor family and that the Kek1/EGFR interaction inhibits growth factor binding, receptor autophosphorylation and Erk1/2 activation in response to EGF. Finally, in vivo experiments show that Kek1 expression potently suppresses the growth of mouse mammary tumor cells derived from aberrant ErbB receptors activation, but does not interfere with the growth of tumor cells derived from activated Ras. Our results underscore the possibility that Kek1 may be used experimentally to inhibit ErbB receptors and point to the possibility that, as yet uncharacterized, mammalian transmembrane LRR proteins might act as modulators of growth factor signalling. PMID- 12900465 TI - Polemic and public health. PMID- 12900468 TI - Vide VIDUS. PMID- 12900469 TI - Training on the wards. PMID- 12900471 TI - Training on the wards. PMID- 12900473 TI - An Oasys for occupational asthma. PMID- 12900474 TI - Training on the wards. PMID- 12900475 TI - COX-2 inhibitors and type 4 error. PMID- 12900476 TI - Effects of thrombolysis for acute stroke in patients with pre-existing disability. AB - BACKGROUND: Thrombolysis for acute stroke is beneficial in selected patients. Because clinical trials generally exclude patients with pre-existing disability, this subgroup of patients has not been studied. We examined the outcomes after thrombolysis of patients with and without disability before their stroke. METHODS: We prospectively followed 112 consecutive patients with acute ischemic stroke who were given intravenous thrombolysis treatment according to published protocols. Three-month outcomes of the patients with pre-existing disability (defined as a prestroke score of 2 or more on the modified Rankin scale [MRS]) were compared with those of patients without pre-existing disability (defined as a prestroke MRS score of 0 or 1) and with those of 168 patients similarly treated in the National Institute of Neurological Disorders and Stroke trial. RESULTS: At 3 months after the stroke, patients with pre-existing disability (21% of the 112) had a higher mortality rate than those without (33% v. 14%) (odds ratio 3.2, 95% confidence interval 1.0-10.1) and worse function (median MRS score 3 v. 2, p = 0.03). However, there was little difference between the 2 groups in neurologic impairment among the survivors (median score on the National Institutes of Health stroke scale 4 v. 2, p = 0.41) or in the total proportion of those with an MRS score of 0 or 1 or, for those with a prestroke score greater than 1, a return to the prestroke score (42% v. 41%, p = 0.87). INTERPRETATION: Although the true effectiveness of thrombolysis for acute stroke in patients with pre-existing disability is not known, treated patients appear able to return to their prestroke level of function as often as patients without pre-existing disability, despite a significantly higher mortality rate. PMID- 12900477 TI - Risk factors for early-onset group B streptococcal disease in neonates: a population-based case-control study. AB - BACKGROUND: Infection with group B streptococcus (GBS) is a major cause of neonatal illness and death. We examined the antenatal and perinatal risk factors for early-onset GBS disease among neonates. METHODS: We identified cases by population-based surveillance in all microbiology laboratories serving Alberta. A case was defined as any instance of a positive sterile-site GBS culture in an infant born between 1993 and 1997 who was either less than 7 days old or stillborn after 20 weeks' gestation. We randomly selected controls from a computer-compiled list of all hospital births, including stillbirths after 20 weeks' gestation, in Alberta during the study period. To increase power, we chose 5 or 6 control infants born in the same year as each case infant. We reviewed hospital, prenatal clinic and physician health records and, between 1997 and 1999, conducted maternal interviews by telephone. RESULTS: There were no differences between the 90 cases and 489 controls in sociodemographic variables or in many reproductive and behavioural variables. Case infants were more likely than control infants to be of low birth weight (odds ratio [OR] 3.60, 95% confidence interval [CI] 1.68-7.65), to have been delivered preterm (OR 3.89, 95% CI 2.08-7.27), or to have a mother with amnionitis (OR 15.03, 95% CI 5.58-41.89), intrapartum fever (OR 4.65, 95% CI 2.48-8.69) or premature rupture of the membranes (OR 2.39, 95% CI 1.38-4.14). After adjustment for potential confounders, intrauterine fetal monitoring was associated with a more than 2-fold increase in the risk of neonatal GBS disease (OR 2.24, 95% CI 1.22-4.13). INTERPRETATION: Intrauterine fetal monitoring should be added to the list of risk factors in risk-based screening. Since many of the cases had no identifiable maternal risk factors, universal screening for GBS may be appropriate. PMID- 12900478 TI - Colorectal cancer screening in Canada: why not consider nurse endoscopists? PMID- 12900479 TI - New grades for recommendations from the Canadian Task Force on Preventive Health Care. PMID- 12900480 TI - DEET-based insect repellents: safety implications for children and pregnant and lactating women. AB - Reducing the risk of mosquito bites is currently the only way to reduce the risk of West Nile virus infection. Methods for avoiding mosquito bites include limiting the time spent outdoors at dawn and dusk, wearing protective clothing and using an insect repellent. Repellents containing DEET (N,N-diethyl-m toluamide, also known as N,N-diethyl-3-methylbenzamide) are the most effective and most widely used. However, concerns have been raised over the risk of adverse toxic effects, especially in young children and pregnant and lactating women. In this article, we review the available evidence on the effectiveness and safety of DEET-based products. The evidence does not support increased risk in young children. PMID- 12900481 TI - Use of back belts to prevent occupational low-back pain. Recommendation statement from the Canadian Task Force on Preventive Health Care. PMID- 12900482 TI - Do hip protectors help prevent hip fractures? PMID- 12900483 TI - Acute venous stasis and swelling of the lower abdomen and extremities in an infant after circumcision. PMID- 12900485 TI - CYPHER coronary stents and risk of thrombosis. PMID- 12900487 TI - Medical marijuana to sell for 5 dollars per gram. PMID- 12900486 TI - Neurosurgeon takes health care helm in Quebec. PMID- 12900491 TI - Health of the nation: "I'm fine, eh". PMID- 12900495 TI - Has my patient achieved adequate myocardial reperfusion? PMID- 12900496 TI - Aortic dissection: new frontiers in diagnosis and management: Part I: from etiology to diagnostic strategies. PMID- 12900497 TI - Cardiology patient pages. Ambulatory electrocardiographic recording. PMID- 12900498 TI - Images in cardiovascular medicine. Detection of a vulnerable coronary plaque: a treatment dilemma. PMID- 12900500 TI - Coarse-grained microscopic model of glass formers. AB - We introduce a coarse-grained model for atomic glass formers. Its elements are physically motivated local microscopic dynamical rules parameterized by observables. Results of the model are established and used to interpret the measured behaviors of supercooled fluids approaching glass transitions. The model predicts the presence of a crossover from hierarchical super-Arrhenius dynamics at short length scales to diffusive Arrhenius dynamics at large length scales. This prediction distinguishes our model from other theories of glass formers and can be tested by experiment. PMID- 12900499 TI - Paclitaxel-eluting stents come out winners again. PMID- 12900501 TI - Generation of systemin signaling in tobacco by transformation with the tomato systemin receptor kinase gene. AB - The tomato systemin receptor, SR160, a plasma membrane-bound, leucine-rich repeat receptor kinase that signals systemic plant defense, and the brassinolide (BL) receptor, BRI1, that regulates developmental processes, have been shown recently to have identical amino acid sequences. We report herein that tobacco, a solanaceous species that does not express a systemin precursor gene nor responds to systemin, when transformed with the SR160 receptor gene, expresses the gene in suspension-cultured cells, evidenced by mRNA and protein analyses and photoaffinity-labeling experiments. Additionally, systemin induced an alkalinization response in the transgenic tobacco cells similar to that found in tomato cells, but not in WT cells. The gain in function in tobacco cells indicates that early steps of the systemin signaling pathway found in tomato are present in tobacco cells. A tomato line, cu-3, in which a mutation in the BRI1 gene has rendered the plant nonfunctional in BL signaling, exhibits a severely reduced response to systemin. In leaves of WT tomato plants, BL strongly and reversibly antagonized systemic signaling by systemin. The results suggest that the systemin-mediated systemic defense response may have evolved in some solanaceous species by co-opting the BRI1 receptor and associated components for defense signaling. PMID- 12900502 TI - Enhanced diffusion-edited NMR spectroscopy of mixtures using chromatographic stationary phases. AB - We introduce an analytical method that combines in one pot the advantages of column chromatography separation and NMR structural analysis. The separation of the NMR spectra of the components of a mixture can be achieved according to their apparent diffusion rates [James, T. L. and McDonald, G. G. (1973) J. Magn. Reson. 58, 58-61]. We show that the separation of the spectral components, corresponding to single molecular species, can be enhanced by order of magnitudes upon addition of a typical stationary phase used in HPLC. The solid phase imbibed by the mixture for analysis is an heterogeneous ensemble, so that solid-state NMR methods (high-resolution magic angle spinning) are necessary to recover high resolution spectra. We demonstrate applications of this combination of high resolution magic angle spinning and NMR diffusometry on test mixtures for direct (silica gel) and inverse (C18) columns. However, many common chromatographic supports available for HPLC should be readily adaptable for use with this technique. PMID- 12900503 TI - Minimal haplotype tagging. AB - The high frequency of single-nucleotide polymorphisms (SNPs) in the human genome presents an unparalleled opportunity to track down the genetic basis of common diseases. At the same time, the sheer number of SNPs also makes unfeasible genome wide disease association studies. The haplotypic nature of the human genome, however, lends itself to the selection of a parsimonious set of SNPs, called haplotype tagging SNPs (htSNPs), able to distinguish the haplotypic variations in a population. Current approaches rely on statistical analysis of transmission rates to identify htSNPs. In contrast to these approximate methods, this contribution describes an exact, analytical, and lossless method, called BEST (Best Enumeration of SNP Tags), able to identify the minimum set of SNPs tagging an arbitrary set of haplotypes from either pedigree or independent samples. Our results confirm that a small proportion of SNPs is sufficient to capture the haplotypic variations in a population and that this proportion decreases exponentially as the haplotype length increases. We used BEST to tag the haplotypes of 105 genes in an African-American and a European-American sample. An interesting finding of this analysis is that the vast majority (95%) of the htSNPs in the European-American sample is a subset of the htSNPs of the African American sample. This result seems to provide further evidence that a severe bottleneck occurred during the founding of Europe and the conjectured "Out of Africa" event. PMID- 12900504 TI - Regulation of the murine renal vitamin D receptor by 1,25-dihydroxyvitamin D3 and calcium. AB - Renal vitamin D receptor (VDR) is required for 1,25-dihydroxyvitamin D3 [1,25(OH)2D3]-induced renal reabsorption of calcium and for 1,25(OH)2D3-induced 1,25(OH)2D3 24-hydroxylase. The long-term effect of vitamin D and dietary calcium on the expression of renal VDR was examined in the nonobese diabetic mouse. Vitamin D-deficient and vitamin D-replete mice were maintained on diets containing 0.02%, 0.25%, 0.47%, and 1.20% calcium with or without 50 ng of 1,25(OH)2D3 per day. Vitamin D-replete mice on a 1.20% calcium diet had renal VDR levels of 165 fmol/mg protein. Calcium restriction caused renal VDR levels to decrease to <30 fmol/mg protein in vitamin D-deficient mice and to approximately 80 fmol/mg protein in vitamin D-replete mice. When dietary calcium was present, 50 ng of 1,25(OH)2D3 elevated the VDR levels 2- to 10-fold, depending on vitamin D status and the level of calcium. In the absence of either vitamin D or calcium, the VDR mRNA was expressed at a basal level. 1,25(OH)2D3 supplementation caused relative VDR mRNA to increase 8- to 10-fold in the vitamin D-deficient mouse when dietary calcium was available. This increase was completely absent in the calcium restricted mice. This in vivo study demonstrates that 1,25(OH)2D3 and calcium are both required for renal VDR mRNA expression above a basal level, furthering our understanding of the complex regulation of renal VDR by 1,25(OH)2D3 and calcium. PMID- 12900505 TI - A gene expression-based method to diagnose clinically distinct subgroups of diffuse large B cell lymphoma. AB - To classify cancer specimens by their gene expression profiles, we created a statistical method based on Bayes' rule that estimates the probability of membership in one of two cancer subgroups. We used this method to classify diffuse large B cell lymphoma (DLBCL) biopsy samples into two gene expression subgroups based on data obtained from spotted cDNA microarrays. The germinal center B cell-like (GCB) DLBCL subgroup expressed genes characteristic of normal germinal center B cells whereas the activated B cell-like (ABC) DLBCL subgroup expressed a subset of the genes that are characteristic of plasma cells, particularly those encoding endoplasmic reticulum and golgi proteins involved in secretion. We next used this predictor to discover these subgroups within a second set of DLBCL biopsies that had been profiled by using oligonucleotide microarrays [Shipp, M. A., et al. (2002) Nat. Med. 8, 68-74]. The GCB and ABC DLBCL subgroups identified in this data set had significantly different 5-yr survival rates after multiagent chemotherapy (62% vs. 26%; P < or = 0.0051), in accord with analyses of other DLBCL cohorts. These results demonstrate the ability of this gene expression-based predictor to classify DLBCLs into biologically and clinically distinct subgroups irrespective of the method used to measure gene expression. PMID- 12900506 TI - The VirD2 pilot protein of Agrobacterium-transferred DNA interacts with the TATA box-binding protein and a nuclear protein kinase in plants. AB - The bacterial virulence protein VirD2 plays an important role in nuclear import and chromosomal integration of Agrobacterium-transferred DNA in fungal, plant, animal, and human cells. Here we show that in nuclei of alfalfa cells, VirD2 interacts with and is phosphorylated by CAK2Ms, a conserved plant ortholog of cyclin-dependent kinase-activating kinases. CAK2Ms binds to and phosphorylates the C-terminal regulatory domain of RNA polymerase II largest subunit, which can recruit the TATA box-binding protein. VirD2 is found in tight association with the TATA box-binding protein in vivo. These results indicate that recognition of VirD2 is mediated by widely conserved nuclear factors in eukaryotes. PMID- 12900508 TI - Rhythmic expression of clock and clock-controlled genes in the rat oviduct. AB - The rhythmic expression of clock and clock-controlled genes in the rat oviduct was investigated by real time RT-PCR. per1, per2, Clock, Bmal1, cry1 and cry2 were all expressed in the oviduct. With 4-hourly sampling over 24 h in a normal photoperiod, analysis of variance indicated that per2 and Bmal1 had highly significant sinusoidal-like changes with an amplitude of 3- and 10-fold respectively. Of the other clock genes, per1 and cry1 had non-significant rhythm amplitudes of 2.5- and 1.8-fold respectively. Using the same experimental approach the rhythmic expression of Bmal1, per1 and per2 mRNA in the liver was found to be highly significant with amplitudes of approximately 20-, 10- and 5 fold respectively. The expression of the clock-controlled transcription factors DBP and Rev-erb alpha showed significant rhythmicity in the oviduct with 5-fold changes in amplitude for both genes. Plasminogen activator inhibitor-1 (PAI-1), which has been implicated in oviduct function during the preimplantation period, also had a significant rhythm of expression (2.5-fold amplitude), peaking at the same time as the other clock-controlled genes, DBP and Rev-erb alpha. These results show for the first time that the female reproductive tract is inherently rhythmic and suggests that the developing embryo may be subjected to rhythmic changes in the environment created by the oviduct during transition to the uterus. PMID- 12900507 TI - Dissociation of amyloid fibrils of alpha-synuclein and transthyretin by pressure reveals their reversible nature and the formation of water-excluded cavities. AB - Protein misfolding and aggregation have been linked to several human diseases, including Alzheimer's disease, Parkinson's disease, and systemic amyloidosis, by mechanisms that are not yet completely understood. The hallmark of most of these diseases is the formation of highly ordered and beta-sheet-rich aggregates referred to as amyloid fibrils. Fibril formation by WT transthyretin (TTR) or TTR variants has been linked to the etiology of systemic amyloidosis and familial amyloid polyneuropathy, respectively. Similarly, amyloid fibril formation by alpha-synuclein (alpha-syn) has been linked to neurodegeneration in Parkinson's disease, a movement disorder characterized by selective degeneration of dopaminergic neurons in the substantia nigra. Here we show that consecutive cycles of compression-decompression under aggregating conditions lead to reversible dissociation of TTR and alpha-syn fibrils. The high sensitivity of amyloid fibrils toward high hydrostatic pressure (HHP) indicates the existence of packing defects in the fibril core. In addition, through the use of HHP we are able to detect differences in stability between fibrils formed from WT TTR and the familial amyloidotic polyneuropathy-associated variant V30M. The fibrils formed by WT alpha-syn were less susceptible to pressure denaturation than the Parkinson's disease-linked variants, A30P and A53T. This finding implies that fibrils of alpha-syn formed from the variants would be more easily dissolved into small oligomers by the cellular machinery. This result has physiological importance in light of the current view that the pathogenic species are the small aggregates rather the mature fibrils. Finally, the HHP-induced formation of fibrils from TTR is relatively fast (approximately 60 min), a quality that allows screening of antiamyloidogenic drugs. PMID- 12900509 TI - Molecular basis of the TaqI p49a,f polymorphism in the DYS1 locus containing DAZ genes. AB - The previously described TaqI p49a,f polymorphism at the DYS1 locus in the non recombinant part of the Y chromosome is widely exploited to investigate many facets of human population genetics. It has been shown that the DYS1 locus corresponds to the four DAZ genes in the AZF-c region of the Y chromosome. As the DNA sequence of the DAZ genes is known in its entirety, it is now possible to establish correspondences between the main Southern polymorphic TaqI bands (A, C, D, F and I) at the DYS1 locus and TaqI fragments deduced from the sequence, by way of comparing band sizes and sequence homologies between hybridized fragments. Transitions between polymorphic forms for each variable TaqI fragment can be explained regarding the restriction maps, by postulating a parsimonious number of TaqI site losses during human evolution. Most of the codon changes caused by TaqI site losses located in the exons of the four DAZ genes have potentially high selective values. PMID- 12900510 TI - No AZF deletion in 160 patients with testicular germ cell neoplasia. AB - Testicular germ cell cancer is aetiologically linked to genital malformations and male infertility and is most probably caused by a disruption of embryonic programming and gonadal development during fetal life. In some cases, germ cell neoplasia is associated with a relative reduction of Y chromosomal material (e.g. 45,X/46,XY) or other abnormalities of the Y chromosome. The euchromatic long arm of the human Y chromosome (Yq11) contains three azoospermia factors (AZFa, AZFb, AZFc) functionally important in human spermatogenesis. Microdeletions encompassing one of these three AZF loci result in the deletion of multiple genes normally expressed in testis tissue and are associated with spermatogenic failure. The aim of our study was to investigate whether AZF microdeletions, in addition to causing infertility, predispose also to germ cell neoplasia, since subjects with poor spermatogenesis have an increased risk of testicular cancer. We screened for putative deletions of AZF loci on the Y chromosome in DNA isolated from white blood cells of 160 Danish patients with testicular germ cell neoplasia. Interestingly, although AZF microdeletions are found frequently in patients with idiopathic infertility, in all cases studied of testicular germ cell cancer the Yq region was found to be intact. We conclude that the molecular aetiology of testicular germ cell neoplasia of the young adult type most likely does not involve the same pathways as male infertility caused by AZF deletions. Malignant transformation of germ cells is thus caused by the dysfunction of some other genes that still need to be identified. PMID- 12900511 TI - ICSI-generated mouse zygotes exhibit altered calcium oscillations, inositol 1,4,5 trisphosphate receptor-1 down-regulation, and embryo development. AB - ICSI bypasses not only fusion of the gametes but also a series of signalling events that occur in the sperm prior to and during interaction with the oocyte's vestments. The effect of this altered encounter of the gametes on the fertilization-associated intracellular calcium ([Ca2+]i) oscillations has not been thoroughly investigated. Here, ICSI and IVF were performed using gametes from two mouse strains, B6D2F1 and CD1, and in-vitro development, pattern of [Ca2+]i oscillations and down-regulation of inositol 1,4,5-trisphosphate receptor 1 (IP3R-1) in the produced embryos were compared. ICSI and IVF resulted in comparable rates of activation and pre-implantation development. However, ICSI generated zygotes cleaved at a slower rate, had lower cell numbers and lower hatching rates. The deleterious effects of ICSI could not be exclusively attributed to the injury by the injection since sham-injected IVF zygotes only exhibited delayed progression to the blastocyst stage. ICSI and IVF induced similar initial [Ca2+]i responses, although ICSI zygotes exhibited shorter durations of [Ca2+]i oscillations and showed diminished degradation of IP3R-1. Importantly, sperm manipulation affected the pattern of oscillations, which further decreased pre-implantation developmental rates. Our results demonstrate that ICSI-induced [Ca2+]i responses are not equivalent to those initiated by IVF and that this may have developmental consequences. PMID- 12900512 TI - Macrophage inhibitory cytokine 1 in fetal membranes and amniotic fluid from pregnancies with and without preterm labour and premature rupture of membranes. AB - The placenta and fetal membranes are the site of expression of macrophage inhibitory cytokine (MIC-1), a member of the transforming growth factor (TGF) beta superfamily. We hypothesized that MIC-1 may act as an immune regulator in pregnancy complications associated with intrauterine inflammation. Decidual cells, chorionic trophoblasts and amnion epithelial cells were identified by immunohistochemistry as the predominant MIC-1-containing cell type in term membranes. Amnion and choriodecidual explants all produced MIC-1 in culture, the latter having the greatest production rate (206 +/- 74.5 pg/mg tissue/24 h, n=6; mean +/- SEM). Production was not responsive to stimulation by pro-inflammatory cytokines. MIC-1 was detectable in 217 transabdominal amniotic fluid (AF) samples taken from 15 to 41 weeks gestation, concentrations ranging from 0.9-51.1 ng/ml. AF MIC-1 concentrations in pregnancies with premature rupture of membranes (PROM) or preterm labour, either with or without microbial invasion of the amniotic cavity, were not significantly different from those delivered at term either with or without labour. Treatment with MIC-1 (0.25-25 ng/ml) did not alter production of interleukin-6 or -8 by amnion or choriodecidual cells in vitro. We conclude that AF MIC-1 is derived from the fetal membranes and decidua, but that MIC-1 is unlikely to be involved in the pathophysiology of preterm birth or PROM. PMID- 12900513 TI - Fibroids display an anti-angiogenic gene expression profile when compared with adjacent myometrium. AB - The aetiology of uterine fibroids remains unknown, despite causing significant gynaecological morbidity. Fibroids have a reduced microvascular density when compared with adjacent myometrial tissue. The aim of this study was to identify genes with differential expression between fibroid and adjacent normal myometrium, particularly genes with a role in angiogenesis. Total RNA was extracted from fibroid/myometrium pairs from 12 hysterectomy specimens, and used to perform 24 cDNA microarrays. There were 10,500 genes screened on each microarray for differential expression. Analysis of expression data was carried out using multiple t-tests, as well as a novel class prediction algorithm (GeneRaVE). The differential gene expression of selected genes was confirmed by quantitative 'real time' RT-PCR. Selected genes with a role in angiogenesis were further analysed for expression in isolated cell populations of endothelial cells (fibroid and myometrium) and smooth muscle cells (fibroid and myometrium), to see if their expression was confined to particular cell types. Twenty-five genes with differential gene expression between fibroid and myometrium were identified. Insulin-like growth factor-2, endothelin A receptor, connective tissue growth factor (CTGF), cysteine-rich angiogenic inducer 61 (CYR61) and collagen 4alpha2 (COL4A2) were confirmed by RT-PCR. CTGF and CYR61, both angiogenesis promoters, were reduced in expression relative to myometrium. COL4A2, the precursor for an angiogenesis inhibitor, canstatin, was increased relative to myometrium. These three genes display an anti-angiogenic expression profile in fibroids relative to myometrium. These findings may explain the reduced microvascular density seen in fibroids relative to myometrium. PMID- 12900514 TI - Expression of membrane-bound HLA-G at the maternal-fetal interface is not associated with pregnancy maintenance among patients with idiopathic recurrent pregnancy loss. AB - Recurrent pregnancy loss (RPL) has many known aetiologies. However, using current diagnostic testing, a large fraction of recurrent pregnancy losses remain unexplained. Many of these may have immune underpinnings. HLA-G is a non classical MHC class I product whose fairly restricted expression at the maternal fetal interface suggests a role in successful embryonic implantation and/or subsequent pregnancy maintenance. The study of immune-mediated RPL should be enhanced by comparing groups of idiopathic RPL patients with normal fetal chromosomes to RPL patients with known chromosomal abnormalities in their index pregnancies. We hypothesized that if alteration of HLA-G expression at the maternal-fetal interface were associated with immune-mediated RPL, such changes might be detectable using these comparisons. HLA-G protein expression at the maternal-fetal interface in maternal and gestational age-matched women with history of idiopathic RPL and normal male fetuses were compared with expression in RPL patients with known fetal trisomy 16 in their index pregnancy. We detected no significant quantitative differences in the levels of HLA-G between these groups of RPL patients. Within the limitations of this study, we conclude that HLA-G expression is not a major immunological determinant of pregnancy maintenance among patients with idiopathic RPL. PMID- 12900515 TI - Improving clinical preimplantation genetic diagnosis for cystic fibrosis by duplex PCR using two polymorphic markers or one polymorphic marker in combination with the detection of the DeltaF508 mutation. AB - Cystic fibrosis (CF) is an autosomal recessive disease characterized by obstruction and chronic infection of the respiratory tract and pancreatic insufficiency. The first preimplantation genetic diagnosis (PGD) for CF was carried out in 1992. At our centre the first cycle was performed in 1993. However, the number of known CF mutations is >1000, so developing mutation specific PCR protocols for PGD is unfeasible. This is why a number of marker based duplex PCRs were developed at the single cell level. A duplex PCR of a mutation and one or two microsatellites is not only a diagnostic tool, but it can also be used as a control for allele drop-out and contamination. During PGD, embryos obtained in vitro are analysed for the presence or absence of a particular genetic disease, after which only embryos shown to be free of this disease are returned to the mother. In total, 22 PGD cycles with duplex PCR (IVS8CA/IVS17BTA, DeltaF508/IVS8CA, DeltaF508/IVS17BTA and D7S486/D7S490) were carried out in 16 couples, which resulted in four ongoing pregnancies and one miscarriage. PMID- 12900516 TI - Differentiation of effector/memory Vdelta2 T cells and migratory routes in lymph nodes or inflammatory sites. AB - Vdelta2 T lymphocytes recognize nonpeptidic antigens without presentation by MHC molecules and mount both immediate effector functions and memory responses after microbial infection. However, how Vdelta2 T cells mediate different facets of a memory response remains unknown. Here, we show that the expression of CD45RA and CD27 antigens defines four subsets of human Vdelta2 T cells with distinctive compartmentalization routes. Naive CD45RA+CD27+ and memory CD45RA-CD27+ cells express lymph node homing receptors, abound in lymph nodes, and lack immediate effector functions. Conversely, memory CD45RA-CD27- and terminally differentiated CD45RA+CD27- cells, which express receptors for homing to inflamed tissues, are poorly represented in the lymph nodes while abounding at sites of inflammation, and display immediate effector functions. These observations and additional in vitro experiments indicate a lineage differentiation pattern for human Vdelta2 T cells that generates naive cells circulating in lymph nodes, effector/memory cells patrolling the blood, and terminally differentiated effector cells residing in inflamed tissues. PMID- 12900517 TI - Peroxisome proliferator-activated receptors alpha and gamma down-regulate allergic inflammation and eosinophil activation. AB - Allergic asthma is characterized by airway hyperresponsiveness, eosinophilia, and mucus accumulation and is associated with increased IgE concentrations. We demonstrate here that peroxisome proliferator-activated receptors (PPARs), PPAR alpha and PPAR-gamma, which have been shown recently to be involved in the regulation of various cell types within the immune system, decrease antigen induced airway hyperresponsiveness, lung inflammation, eosinophilia, cytokine production, and GATA-3 expression as well as serum levels of antigen-specific IgE in a murine model of human asthma. In addition, we demonstrate that PPAR-alpha and -gamma are expressed in eosinophils and their activation inhibits in vitro chemotaxis and antibody-dependent cellular cytotoxicity. Thus, PPAR-alpha and gamma (co)agonists might be of therapeutic interest for the regulation of allergic or inflammatory reactions by targeting both regulatory and effector cells involved in the immune response. PMID- 12900518 TI - The chymase, mouse mast cell protease 4, constitutes the major chymotrypsin-like activity in peritoneum and ear tissue. A role for mouse mast cell protease 4 in thrombin regulation and fibronectin turnover. AB - To gain insight into the biological role of mast cell chymase we have generated a mouse strain with a targeted deletion in the gene for mast cell protease 4 (mMCP 4), the mouse chymase that has the closest relationship to the human chymase in terms of tissue localization and functional properties. The inactivation of mMCP 4 did not affect the storage of other mast cell proteases and did not affect the number of mast cells or the mast cell morphology. However, mMCP-4 inactivation resulted in complete loss of chymotryptic activity in the peritoneum and in ear tissue, indicating that mMCP-4 is the main source of stored chymotrypsin-like protease activity at these sites. The mMCP-4 null cells showed markedly impaired ability to perform inactivating cleavages of thrombin, indicating a role for mMCP 4 in regulating the extravascular coagulation system. Further, a role for mMCP-4 in connective tissue remodeling was suggested by the inability of mMCP-4 null peritoneal cells to process endogenous fibronectin. PMID- 12900519 TI - Gamma delta T cells provide an early source of interferon gamma in tumor immunity. AB - Interferon (IFN)-gamma is necessary for tumor immunity, however, its initial cellular source is unknown. Because gammadelta T cells primarily produce this cytokine upon activation, we hypothesized that they would provide an important early source of IFN-gamma in tumor immunosurveillance. To address this hypothesis, we first demonstrated that gammadelta T cell-deficient mice had a significantly higher incidence of tumor development after challenge with a chemical carcinogen methylcholanthrene (MCA) or inoculation with the melanoma cell line B16. In wild-type mice, gammadelta T cells were recruited to the site of tumor as early as day 3 after inoculation, followed by alphabeta T cells at day 5. We then used bone marrow chimeras and fetal liver reconstitutions to create mice with an intact gammadelta T cell repertoire but one that was specifically deficient in the capacity to produce IFN-gamma. Such mice had a higher incidence of tumor development, induced either with MCA or by inoculation of B16 melanoma cells, compared with mice with IFN-gamma-competent gammadelta T cells. Moreover, genetic deficiency of gammadelta T cells resulted in impaired IFN-gamma production by tumor antigen-triggered alphabeta T cell upon immunization with tumor lysate. These results demonstrate that gammadelta T cells can play a necessary role in tumor immunity through provision of an early source of IFN-gamma that in turn may regulate the function of tumor-triggered alphabeta T cells. PMID- 12900520 TI - CSF-induced and HIV-1-mediated distinct regulation of Hck and C/EBPbeta represent a heterogeneous susceptibility of monocyte-derived macrophages to M-tropic HIV-1 infection. AB - Granulocyte/macrophage colony-stimulating factor (GM-CSF)-induced monocyte derived macrophages (GM-MPhi) are permissive to M-tropic HIV-1 entry, but inhibit viral replication at posttranscriptional and translational levels, whereas M-CSF induced macrophages (M-MPhi) produce a large amount of HIV-1. M-MPhi express a high level of Hck and a large isoform of C/EBPbeta, and HIV-1 infection increases the expression of Hck but not of C/EBPbeta. GM-MPhi express a high level of C/EBPbeta and a low level of Hck, and HIV-1 infection drastically increases the expression of a short isoform of C/EBPbeta but decreases that of Hck. Treatment of M-MPhi with antisense oligonucleotide for Hck (AS-Hck) not only suppresses the expression of Hck, but also stimulates the induction of the short isoform of C/EBPbeta and inhibits the viral replication. Treatment of GM-MPhi with a moderate amount of AS-C/EBPbeta not only inhibits the expression of the small isoform of C/EBPbeta preferentially, but also stimulates the induction of Hck and stimulates the virus production at a high rate. These results suggest that CSF induced and HIV-1-mediated distinct regulation of Hck and small isoform of C/EBPbeta represent the heterogeneous susceptibility of tissue MPhi to HIV-1 infection, and the regulation of Hck and C/EBPbeta are closely related and these two molecules affect one another. PMID- 12900521 TI - The neurotrophins nerve growth factor, brain-derived neurotrophic factor, neurotrophin-3, and neurotrophin-4 are survival and activation factors for eosinophils in patients with allergic bronchial asthma. AB - Neurotrophins (nerve growth factor [NGF], brain-derived neurotrophic factor [BDNF], neurotrophin [NT]-3, and NT-4) have been observed in elevated concentrations in allergic diseases. Neurotrophin levels are up-regulated endobronchially after allergen challenge. This coincides with an influx of activated eosinophils into the bronchial lumen. These eosinophils have an increased viability and CD69 expression 18 h after segmental allergen provocation (SAP) which is not present in peripheral blood. To investigate whether these observations are related we studied the influence of neurotrophins on eosinophil function in allergic asthma. Incubation with NGF, BDNF, NT-3, or NT-4 caused a significant increase in the viability and CD69 expression of isolated eosinophils from bronchoalveolar lavage fluid (BALF) but not from peripheral blood, suggesting a unique sensitivity of endobronchial eosinophils to neurotrophins. To elucidate the underlying mechanisms expression of the neurotrophin receptors p75NTR, trkA, trkB, and trkC on eosinophils was analyzed by RT-PCR and immunocytology. After SAP expression of all neurotrophin receptors was markedly elevated on eosinophils from BALF. Our findings suggest that neurotrophin mediated activation of bronchial eosinophils might play a role in the regulation of eosinophilic inflammation in allergic asthma. PMID- 12900522 TI - VEGF164-mediated inflammation is required for pathological, but not physiological, ischemia-induced retinal neovascularization. AB - Hypoxia-induced VEGF governs both physiological retinal vascular development and pathological retinal neovascularization. In the current paper, the mechanisms of physiological and pathological neovascularization are compared and contrasted. During pathological neovascularization, both the absolute and relative expression levels for VEGF164 increased to a greater degree than during physiological neovascularization. Furthermore, extensive leukocyte adhesion was observed at the leading edge of pathological, but not physiological, neovascularization. When a VEGF164-specific neutralizing aptamer was administered, it potently suppressed the leukocyte adhesion and pathological neovascularization, whereas it had little or no effect on physiological neovascularization. In parallel experiments, genetically altered VEGF164-deficient (VEGF120/188) mice exhibited no difference in physiological neovascularization when compared with wild-type (VEGF+/+) controls. In contrast, administration of a VEGFR-1/Fc fusion protein, which blocks all VEGF isoforms, led to significant suppression of both pathological and physiological neovascularization. In addition, the targeted inactivation of monocyte lineage cells with clodronate-liposomes led to the suppression of pathological neovascularization. Conversely, the blockade of T lymphocyte mediated immune responses with an anti-CD2 antibody exacerbated pathological neovascularization. These data highlight important molecular and cellular differences between physiological and pathological retinal neovascularization. During pathological neovascularization, VEGF164 selectively induces inflammation and cellular immunity. These processes provide positive and negative angiogenic regulation, respectively. Together, new therapeutic approaches for selectively targeting pathological, but not physiological, retinal neovascularization are outlined. PMID- 12900523 TI - Normal thymocyte negative selection in TRAIL-deficient mice. AB - The molecular basis of thymocyte negative selection, which plays a critical role in establishing and maintaining immunological tolerance, is not yet resolved. In particular, the importance of the death receptor subgroup of the tumor necrosis factor (TNF)-family has been the subject of many investigations, with equivocal results. A recent report suggested that TRAIL was a critical factor in this process, a result that does not fit well with previous studies that excluded a role for the FADD-caspase 8 pathway, which is essential for TRAIL and Fas ligand (FasL) signaling, in negative selection. We have investigated intrathymic negative selection of TRAIL-deficient thymocytes, using four well-established models, including antibody-mediated TCR/CD3 ligation in vitro, stimulation with endogenous superantigen in vitro and in vivo, and treatment with exogenous superantigen in vitro. We were unable to demonstrate a role for TRAIL signaling in any of these models, suggesting that this pathway is not a critical factor for thymocyte negative selection. PMID- 12900524 TI - A conduit system distributes chemokines and small blood-borne molecules through the splenic white pulp. AB - Access to the splenic white pulp is restricted to lymphocytes and dendritic cells. Here we show that movement of molecules from the blood into these confined areas is also limited. Large molecules, such as bovine serum albumin (68 kD), immunoglobulin G (150 kD), and 500 kD dextran are unable to enter the white pulp, whereas smaller blood-borne molecules can directly permeate this compartment. The distribution is restricted to a stromal network that we refer to as the splenic conduit system. The small lumen of the conduit contains collagen fibers and is surrounded in the T cell areas by reticular fibroblasts that express ER-TR7. It also contains the chemokine CCL21. Conversely, in B cell follicles the B cell attracting chemokine CXCL13 was found to be associated with the conduit and absence of ER-TR7+ fibroblasts. These results show heterogeneity of reticular fibroblasts that enfold the conduit system and suggest that locally produced chemokines are transported through and presented on this reticular network. Therefore, the conduit plays a role in distribution of both blood-borne and locally produced molecules and provides a framework for directing lymphocyte migration and organization of the splenic white pulp. PMID- 12900526 TI - Hodgkin's lymphoma: the hazards of success. PMID- 12900525 TI - Toll-like receptor 9-mediated recognition of Herpes simplex virus-2 by plasmacytoid dendritic cells. AB - Plasmacytoid dendritic cells (pDCs) have been identified as a potent secretor of the type I interferons (IFNs) in response to CpG as well as several viruses. In this study, we examined the molecular mechanism of virus recognition by pDCs. First, we demonstrated that the CD11c+Gr-1intB220+ pDCs from mouse bone marrow secreted high levels of IFN-alpha in response to either live or UV-inactivated Herpes simplex virus-2 (HSV-2). Next, we identified that IFN-alpha secretion by pDCs required the expression of the adaptor molecule MyD88, suggesting the involvement of a Toll-like receptor (TLR) in HSV-2 recognition. To test whether a TLR mediates HSV-2-induced IFN-alpha secretion from pDCs, various knockout mice were examined. These experiments revealed a clear requirement for TLR9 in this process. Further, we demonstrated that purified HSV-2 DNA can trigger IFN-alpha secretion from pDCs and that inhibitory CpG oligonucleotide treatment diminished HSV-induced IFN-alpha secretion by pDCs in a dose-dependent manner. The recognition of HSV-2 by TLR9 was mediated through an endocytic pathway that was inhibited by chloroquine or bafilomycin A1. The strict requirement for TLR9 in IFN-alpha secretion was further confirmed by the inoculation of HSV-2 in vivo. Therefore, these results demonstrate a novel mechanism whereby the genomic DNA of a virus can engage TLR9 and result in the secretion of IFN-alpha by pDCs. PMID- 12900527 TI - Intravenous nitroglycerin in the management of retained placenta. AB - Thirty patients in the third stage of delivery complicated by retained placenta were prospectively managed with intravenous nitroglycerin. We studied the effect of nitroglycerin on the hemodynamics and characteristics of the procedure, and also analyzed labor and maternal and neonatal data. Intravenous nitroglycerin effectively aided the extraction of the retained placenta in all cases. The average procedure time was 5.3 +/- 1.1 min, and the dose range was 50-200 microg. Systolic and diastolic blood pressure fell significantly from 111 +/- 7.5 to 103 +/- 6 mm Hg and from 74 +/- 6.7 to 67 +/- 6.6 mm Hg, respectively (p<0.05). Although statistically significant, this was not evident clinically and there were no complications. Intravenous nitroglycerin at a dose of 200 microg or less is safe, effective and predictable in the management of retained placenta and could obviate the need of general anesthesia. PMID- 12900528 TI - Analgesic efficacy of etoricoxib in primary dysmenorrhea: results of a randomized, controlled trial. AB - OBJECTIVE: To determine the efficacy of etoricoxib in the treatment of primary dysmenorrhea. METHODS: Seventy-three women were randomly assigned to receive single oral doses of etoricoxib 120 mg, placebo, or naproxen sodium 550 mg at the onset of moderate to severe pain associated with menses. During 3 consecutive menstrual cycles in this double-blind, 3-period, crossover study, pain intensity and pain relief were assessed over the 24-hour period following dosing, and global ratings of therapy were made at 8 and 24 h after dosing. Tolerability was assessed by spontaneous reports of adverse experiences. RESULTS: Etoricoxib 120 mg provided analgesic efficacy superior to placebo for the primary endpoint, total pain relief over 8 h (TOPAR8, p<0.001), and for all secondary endpoints (p<0.050). The analgesic effect of etoricoxib 120 mg over the first 8 h was similar to that of naproxen sodium 550 mg. All treatments were well tolerated. CONCLUSIONS: Etoricoxib 120 mg provided rapid and sustained analgesia that was superior to placebo and similar to that of naproxen sodium 550 mg. PMID- 12900529 TI - Pancreatic pseudocyst in chronic pancreatitis: endoscopic and surgical treatment. PMID- 12900530 TI - 3-dimensional computed tomography image of a benign gastrocolonic fistula. PMID- 12900531 TI - Pancreatic transection from blunt abdominal trauma: early versus delayed diagnosis and surgical management. AB - BACKGROUND AND AIMS: Pancreatic trauma is relatively uncommon, but carries high morbidity and mortality rates, especially when diagnosis is delayed or inappropriate surgery is attempted. PATIENT MATERIAL: The clinical course and surgical management of 14 patients with distal pancreatic transection or severe laceration with or without main pancreatic duct (MPD) injury caused by blunt abdominal trauma were analyzed in a university teaching hospital. The average age of the 14 patients (12 male, 2 female) was 28.9 years (range 5-56). Six patients had isolated pancreatic trauma, and intra-abdominal and extra-abdominal (mean 0.8) injuries associated with pancreatic transection were seen in the other 8 patients. RESULTS: Nine patients were diagnosed and operated on within the first 24 h. Eight of them underwent transection of the gland with MPD injury; distal pancreatectomy with splenectomy was performed in 3 and without splenectomy in 2, distal pancreatogastrostomy in 1, and - due to associated duodenal laceration and/or contusion of the pancreatic head - pylorus-preserving pancreatoduodenectomy in 2. In 1 case (grade II laceration) only external drainage was necessary. All the patients with early, correctly diagnosed parenchymal and ductal injury survived. Only 1 patient required reoperation due to haemorrhage after pancreatoduodenectomy. The other 5 cases were referred elsewhere after initial treatment, and all of them underwent some kind of external drainage. Three had undetected MPD injury, and in the other 2 cases the parenchymal lesions were either underestimated or missed. All of these cases required subsequent resection (1), internal drainage due to fistula (2), or drainage of developed abscess (2). Three of them had severe septic and pulmonary complications; 1 patient with MPD injury was lost to follow-up. CONCLUSION: Patients requiring delayed surgical intervention after an unsuccessful period of observation or a subsequent operation due to undetected MPD injury demonstrated a higher rate of pancreas-specific mortality and morbidity. PMID- 12900532 TI - Treatment of secondary peritonitis: is a less expensive broad-spectrum antibiotic as effective as a carbapenem? AB - BACKGROUND: In this retrospective study, the possibility of using amoxicillin/clavulanate for empirical therapy of secondary peritonitis as an alternative to imipenem was explored. METHODS: All secondary peritonitis cases at our institution between 1998 and 2000 were included. Susceptibility to imipenem and amoxicillin/clavulanate of microorganisms isolated in peritoneal fluid and success rates were compared. Therapeutic failure was defined as death, necessity of repeated surgical intervention, or clinical deterioration with persistent positive cultures. RESULTS: Seventy-six cases of secondary peritonitis with 156 microorganisms were found. One hundred and forty-nine (98%) were susceptible to imipenem versus 124 (82%) to amoxicillin/clavulanate (p = 0.0001). Thirteen therapeutic failures occurred in 52 patients treated with amoxicillin/clavulanate (25%) versus 3 out of 8 (38%) with imipenem (p = 0.43). The proportion of organisms resistant to amoxicillin/clavulanate in therapeutic failures was greater in nosocomial versus community-acquired secondary peritonitis (p = 0.041). CONCLUSION: Despite its better in vitro bacteriological activity, clinical efficacy of imipenem was identical to amoxicillin/clavulanate. The use of amoxicillin/clavulanate instead of imipenem would save 889 Euro per case. PMID- 12900533 TI - Lymphatic mapping and sentinel node biopsy during laparoscopic gastrectomy for early cancer. AB - BACKGROUND/AIMS: Intraoperative lymphatic mapping and sentinel node (SN) biopsy can potentially be combined with minimally invasive surgery, but there are few reports of laparoscopic lymphatic mapping for gastrointestinal cancer. We examined the feasibility and accuracy of laparoscopic lymphatic mapping in predicting lymph node status in patients with gastric cancer. METHODS: Seventeen patients with gastric cancer invading the mucosal or submucosal layers (T1) underwent laparoscopic gastrectomy with lymphatic mapping between March 2001 and May 2002. The day before surgery, a technetium-99m-labelled tin colloid solution was injected endoscopically around the tumor. Immediately after the pneumoperitoneum, patent blue was injected. Gastrectomy was performed in all patients, and blue-stained or radioactive nodes were defined as SNs. Fresh SNs were immediately processed for frozen-section examination by hematoxylin-eosin (H and E) and immunohistochemical (IHC) staining. All non-SNs harvested from resected specimens were subjected to histological examination with H and E. RESULTS: SNs were detected in all patients by combination of the two kinds of tracers. Three patients had lymph node metastases in their final examination, and SNs in these 3 were operatively diagnosed as positive by H and E or IHC staining. Lymphatic mapping and SN biopsy under laparoscopic surgery were performed with 100% accuracy. CONCLUSION: Our preliminary study shows the feasibility of intraoperative lymphatic mapping in laparoscopic gastrectomy for T1 gastric cancer. PMID- 12900534 TI - Double resection for patients with pancreatic cancer and a second primary renal cell cancer. AB - BACKGROUND: Reports of synchronous or metachronous double kidney-pancreas cancers are very rare. METHODS: We present 2 patients with renal cell carcinoma and synchronous (1 patient) or metachronous (1 patient) primary pancreatic ductal adenocarcinoma. The patients underwent resection for both cancer types with a worthwhile outcome. RESULTS: The appearance of different primaries in an individual may indicate a genetic predisposition to different neoplasms. The study of double primary cancers is important because it might provide understanding of a shared genetic basis of different solid tumors. CONCLUSIONS: The association between these two cancers demands more detailed epidemiological and molecular investigation. From a clinical viewpoint a resectional policy is recommended. PMID- 12900535 TI - Intussusception in adults: a 21-year experience in the university-affiliated emergency center and indication for nonoperative reduction. AB - BACKGROUND: While intussusception is relatively common in children, it is rare in adults. METHODS: We retrospectively reviewed the records of all patients older than 18 years with the diagnosis of intussusception between 1981 and 2001. RESULTS: Eleven patients with surgically or endoscopically proven intussusception were encountered at the University-affiliated emergency center. The patients ranged in age from 19 to 88 years with a mean age of 45 years. Males predominated by a ratio of 7:4. Most patients (82%) presented with symptoms of bowel obstruction. The mean duration of symptoms was 4.5 days with a range of 4 h to 25 days. Correct pre-treatment diagnosis was made in 82% of the patients using abdominal ultrasonography and computed tomography (CT). The causes of intussusception were organic lesions in 64% of the patients, postoperative in 18% and idiopathic in 18%, respectively. 73% of patients had emergency operations, and an attempt at nonoperative reduction was performed and completed successfully in 3 patients with ileo-colic or colonic type of intussusception. There have been no cases of morbidity or mortality in our series and no recurrence has occurred up to the present time. CONCLUSIONS: Abdominal ultrasonography and CT were effective tools for the diagnosis of intussusception. Patients with ileo-colic and colonic intussusception without malignant lesions could be good candidates for nonoperative reduction prior to definitive surgery. PMID- 12900536 TI - Invasive carcinoma after endoscopic ablative therapy for high-grade dysplasia in Barrett's oesophagus. AB - BACKGROUND: Patients with high-grade dysplasia (HGD) in Barrett's oesophagus carry a significant risk of developing adenocarcinoma. Endoscopic mucosal resection (EMR) and photodynamic therapy (PDT) aim at the radical ablation of the dysplastic area. METHODS: We used EMR to resect the macroscopic area of dysplastic mucosa followed by PDT to eliminate residual disease. PDT was performed after oral administration of 5-aminolevulinic acid (ALA, 40 mg/kg), using fractionated illumination 3 and 6 h later with 630 nm light at 100 J/cm(2) through an endoscopic balloon diffuser. RESULTS: We report 2 patients who developed adenocarcinoma shortly after incomplete endoscopic ablation of Barrett's epithelium. In a 61-year-old man with HGD in 8-cm Barrett's segment, HGD persisted 3 months after treatment. The oesophagectomy specimen showed a 2.3 cm pT2N0M0 adenocarcinoma in Barrett's. In a 69-year-old woman with extensive HGD in 5-cm Barrett's, HGD persisted after 3 PDT sessions in 1 year. Adenocarcinoma occurred 6 months after treatment. The oesophagectomy showed a pT1bN0M0 adenocarcinoma and extensive multifocal HGD in Barrett's. CONCLUSIONS: The combination of EMR and PDT may be a promising option for local treatment of patients with HGD in Barrett's oesophagus, provided all dysplastic tissue can be removed. Currently it should be offered only to patients who are willing to participate in a clinical trial with an intensive endoscopic follow-up programme. PMID- 12900537 TI - Preoperative natural killer cell activity as a prognostic factor for distant metastasis following surgery for colon cancer. AB - BACKGROUND/AIMS: To determine whether preoperative natural killer (NK) cell activity has any prognostic significance in colon cancer patients. METHODS: The study population consisted of 140 patients with colon cancer. NK cell activity was determined within 2 weeks before surgery in 128 patients and at the time of diagnosis in the remaining 12 patients who either did not undergo surgery or who underwent palliative surgery only. Disease progression and postoperative prognosis were examined in relation to NK cell activity. RESULTS: Decreases in NK cell activity did not necessarily correspond to tumor stage. In curatively operated stage I-III diseases, preoperative NK cell activity of 20% or less correlated with poor survival. Lower activity was also associated with metachronous distant metastases but not with local recurrences. In particular, more than half the stage III patients with attenuated NK cell activity developed metastases. Multivariate analysis indicated that attenuated NK cell activity was a significant parameter for predicting distant metastasis following curative surgery for colon cancer. CONCLUSION: Preoperative NK cell activity has a significant prognostic value in curatively operated colon cancer, particularly for the development of metachronous distant metastasis in stage III patients. PMID- 12900538 TI - Oral nifedipine in the treatment of chronic anal fissure. AB - BACKGROUND: The purpose of this study was to demonstrate the effect of oral nifedipine on maximal resting anal pressure (MRAP) in healthy volunteers and to evaluate its role in the treatment of chronic anal fissure (CAF). METHODS: MRAP was measured in 10 healthy volunteers and 10 patients with CAF before and after oral nifedipine (20 mg b.i.d.). Patients were assessed on the first visit and every fortnight for measurement of MRAP, pain scores, blood pressure, pulse rate, healing of the fissure and adverse effects. Treatments were continued until healing had occurred or for up to 8 weeks. RESULTS: MRAP values before and after nifedipine were 73.2 and 49.3 cm H2O, respectively, in healthy volunteers (p < 0.001). Nifedipine caused a reduction in mean MRAP from 105.2 to 74.0 cm H2O (p < 0.001) in patients with CAF. Pain scores were significantly reduced after 2 weeks of treatment with nifedipine (p < 0.001) and continued throughout the treatment period. At the end of the study 6 of the 10 patients treated with nifedipine were deemed to be healed (n = 5) or improved (n = 1). Headache occurred in 1 patient. CONCLUSION: We conclude that oral nifedipine is effective in reducing MRAP and should become the first-line treatment for CAF. PMID- 12900539 TI - X chromosome inactivation: theme and variations. AB - My contribution to this special issue on Vertebrate Sex Chromosomes deals with the theme of X chromosome inactivation and its variations. I will argue that the single active X--characteristic of mammalian X dosage compensation--is unique to mammals, and that the major underlying mechanism(s) must be the same for most of them. The variable features reflect modifications that do not interfere with the basic theme. These variations were acquired during mammalian evolution--to solve special needs for imprinting and locking in the inactive state. Some of the adaptations reinforce the basic theme, and were needed because of species differences in the timing of interacting developmental events. Elucidating the molecular basis for the single active X requires that we distinguish the mechanisms essential for the basic theme from those responsible for its variations. PMID- 12900540 TI - X chromosome inactivation, differentiation, and DNA methylation revisited, with a tribute to Susumu Ohno. AB - X chromosome inactivation and DNA methylation are reviewed, with emphasis on the contributions of Susumu Ohno and the predictions made in my 1975 paper (Riggs, 1975), in which I proposed the "maintenance methylase" model for somatic inheritance of methylation patterns and suggested that DNA methylation would be involved in mammalian X chromosome inactivation and development. The maintenance methylase model is discussed and updated to consider methylation patterns in cell populations that have occasional, stochastic methylation changes by de novo methylation or demethylation, either active or passive. The "way station" model for the spread of X inactivation by LINE-1 elements is also considered, and some recent results from my laboratory are briefly reviewed. PMID- 12900541 TI - ICF syndrome cells as a model system for studying X chromosome inactivation. AB - Mutations in the DNMT3B DNA methyltransferase gene cause the ICF immunodeficiency syndrome. The targets of this DNA methyltransferase are CpG-rich heterochromatic regions, including pericentromeric satellites and the inactive X chromosome. The abnormal hypomethylation in ICF cells provides an important model system for determining the relationships between replication time, CpG island methylation, chromatin structure, and gene silencing in X chromosome inactivation. PMID- 12900542 TI - X chromosome inactivation: how human are mice? AB - Mammals perform dosage compensation of X-linked gene products between XY males and XX females by transcriptionally silencing all but one X chromosome per diploid cell, a process called X chromosome inactivation (XCI). XCI involves counting X chromosomes in a cell, random or imprinted choice of one X to remain active, initiation and spread of the inactivation signal in CIS throughout the other X chromosomes, and maintenance of the inactive state of those X chromosomes during cell divisions thereafter. Most of what is known of the molecular mechanisms involved in the different steps of XCI has been studied in the mouse. In this review we compare XCI in mouse and human, and discuss how much of the murine data can be extrapolated to humans. PMID- 12900543 TI - Escape from X inactivation. AB - Although the process of X inactivation in mammalian cells silences the majority of genes on the inactivated X chromosome, some genes escape this chromosome-wide silencing. Genes that escape X inactivation present a unique opportunity to study the process of silencing and the mechanisms that protect some genes from being turned off. In this review, we will discuss evolutionary aspects of escape from X inactivation, in relation to the divergence of the sex chromosomes. Molecular characteristics, expression, and epigenetic modifications of genes that escape will be presented, including their developmental regulation and the implications of chromatin domains along the X chromosome in modeling the escape process. PMID- 12900544 TI - Spontaneous reactivation of the inactive X chromosome in mouse embryonal carcinoma cells. AB - The mouse embryonal carcinoma cell line MC12 carries two X chromosomes, one of which replicates late in S phase and shares properties with the normal inactive X chromosome and, therefore, is considered to be inactivated. Since the hypoxanthine phosphoribosyl transferase (HPRT) gene on the active X chromosome is mutated (HPRT(NDASH;)), MC12 cells lack HPRT activity. After subjecting MC12 cells to selection in HAT medium, however, a number of HAT-resistant clones (HAT(R)) appeared. The high frequency of HAT resistance (3.18 x 10(-4)) suggested reactivation of HPRT(PLUS;) on the inactive X chromosome rather than reversion of HPRT(NDASH;). Consistent with this view, cytological analyses showed that the reactivation occurred over the length of the inactive X chromosome in 11 of 20 HAT(R) clones isolated. The remaining nine clones retained a normal heterochromatic inactive X chromosome. The spontaneous reactivation rate of the HPRT(PLUS;) on the inactive X chromosome was relatively high (1.34 x 10(-6)) and comparable to that observed for XIST-deleted somatic cells (Csankovszki et al., 2001), suggesting that the inactivated state is poorly maintained in MC12 cells. PMID- 12900545 TI - Nonrandom X chromosome inactivation in mouse embryos carrying Searle's T(X;16)16H translocation visualized using X-linked LACZ and GFP transgenes. AB - Only the morphologically normal X chromosome is inactivated in female mice heterozygous for Searle's X-autosome translocation, T(X;16)16H. Here we performed a visual study of the primary and secondary events that culminate in the completely nonrandom inactivation of the X in female embryos having this translocation. The data we have obtained so far indicate that the initial choice of the future inactive X chromosome is biased, with the degree of skewing somewhere between 70:30% and 90:10% in favor of the morphologically normal X chromosome. The majority of genetically unbalanced cells that inactivate a translocated X chromosome are quickly eliminated from the embryo proper by E8.5, although the survival of such cells is sporadically observed thereafter. The initial nonrandom choice demonstrated in this study supports the contention that the T(X;16)16H translocation disrupts one of the loci involved in the randomness of the choice of the future inactive X chromosome. Although the HMG-LACZ transgene in H253 stock mice is an excellent marker of X chromosome inactivation, the present study suggests that it is infrequently de-repressed on the inactive X chromosome. PMID- 12900546 TI - Antisense regulation in X inactivation and autosomal imprinting. AB - The regulation of epigenetic phenomena by elements encoding antisense RNA's is one of the most rapidly emerging themes in mammalian gene expression. Such regulation is epitomized by X chromosome inactivation (XCI) and autosomal imprinting. In XCI, TSIX serves as an antisense regulator of XIST, the silencer element for XCI which itself makes a non-coding transcript. Numerous antisense transcripts have also been discovered in autosomally imprinted loci, including the IGF2R/AIR locus, the Prader-Willi/Angelman Syndrome (PWS/AS) locus, and the Beckwith-Wiedemann Syndrome (BWS) locus. How these antisense elements regulate XCI and imprinting remains unsolved. However, various structural and functional similarities among them imply the possibility of shared mechanism. Among the most interesting are the antagonistic relationship between sense and antisense loci and the initiation of antisense transcripts within imprinting centers. This article reviews the latest developments in antisense regulation in XCI and autosomal imprinting and speculates on molecular means by which antisense genes can regulate silencing in mammals. PMID- 12900547 TI - Differential patterns of histone methylation and acetylation distinguish active and repressed alleles at X-linked genes. AB - In female mammals, one of the two X chromosomes is inactivated to compensate for the difference in dosage of X-linked genes between males and females. X inactivation involves sequential alterations to the chromatin that ultimately lead to the transcriptional repression of genes on the X chromosome. Here, histone methylation and acetylation along X-linked genes are investigated by chromatin immunoprecipitation (ChIP) of adult fibroblast cell lines. At PGK1 and HPRT, chromatin on the active X chromosome reveals H3 lysine 4 methylation and acetylation of histones H3 and H4. These modifications are absent on the repressed allele, which is marked by H3 lysine 9 methylation. On the expressed allele of XIST (on the inactive X chromosome), we found that H3 acetylation is confined to the promoter, whereas H3 lysine 4 methylation and H4 acetylation are present along the entire gene. On the repressed XIST allele, in contrast, the promoter and gene exhibit H3 lysine 9 methylation. At only 1.5 kb upstream of the XIST gene, chromatin on the inactive X chromosome has strongly reduced levels of H4 acetylation and is marked by both H3 lysine 9 and H3 lysine 4 methylation. These data demonstrate that patterns of histone methylation and acetylation are distinct along and upstream of XIST and suggest that the inactive X chromatin configuration occurs at a region close to the 5' end of the gene. PMID- 12900548 TI - Integrated kinetics of X chromosome inactivation in differentiating embryonic stem cells. AB - Inactivation of the X chromosome during early female development and the subsequent maintenance of this transcriptionally inert state through countless cell divisions remain a paradigm for epigenetic regulation in mammals. Nevertheless, the exact mechanisms underlying this chromosome-wide silencing process remain unclear. Using differentiating female embryonic stem (ES) cells as a model system, we recently found that histone H3 tail modifications are among the earliest known chromatin changes in the X inactivation process, appearing as soon as Xist RNA accumulates on the X chromosome, but prior to transcriptional silencing of X-linked genes (Heard et al., 2001). In this report we present an integrated analysis of the sequence of early events and chromatin modifications underlying X inactivation in differentiating female ES cells. We have extended our previous analysis concerning changes in histone tail modification states. We find that the hypomethylation of Arg-17 and that of Lys-36 on histone H3 also characterize the inactive X chromosome, and that these profiles show a similarly early onset during the initiation of X inactivation. In addition, we have investigated the kinetics of the shift in replication timing of the X chromosome undergoing inactivation. This event occurs slightly later than Xist RNA coating and the chromatin modifications. Finally, from an early stage in the X inactivation process, characteristic histone modification patterns can be found on the X chromosome at mitosis, suggesting that they represent true epigenetic marks of the inactive state. PMID- 12900549 TI - Overall DNA methylation and chromatin structure of normal and abnormal X chromosomes. AB - DNA methylation patterns were studied at the chromosome level in normal and abnormal X chromosomes using an anti-5-methylcytosine antibody. In man, except for the late-replicating X of female cells, the labeled chromosome structures correspond to R- and T-bands and heterochromatin. Depending on the cell type, the species, and cell culture conditions, the late-replicating X in female cells appears to be more or less undermethylated. Under normal conditions, the only structures that remain methylated on the X chromosomes correspond to pseudoautosomal regions, which harbor active genes. Thus, active genes are usually hypomethylated but are located in methylated chromatin. Structural rearrangements of the X chromosome, such as t(X;X)(pter;pter), induce a Turner syndrome-like phenotype that is inconsistent with the resulting triple-X constitution. This suggests a position effect controlling gene inactivation. The derivative chromosomes are always late replicating, and their duplicated short arms, which harbor pseudoautosomal regions, replicate later than the normal late replicating X chromosomes. The compaction or condensation of this segment is unusual, with a halo of chromatin surrounding a hypocondensed chromosome core. The chromosome core is hypomethylated, but the surrounding chromatin is slightly labeled. Thus, unusual DNA methylation and chromatin condensation are associated with the observed position effect. This strengthens the hypothesis that DNA methylation at the chromosome level is associated with both chromatin structure and gene expression. PMID- 12900550 TI - Ectopic XIST transcripts in human somatic cells show variable expression and localization. AB - XIST encodes a functional RNA that is expressed exclusively from the inactive X in female mammals and is required for the silencing of most of the genes on the chromosome. XIST transcripts remain in the nucleus, and their specific localization to the inactive X is important for silencing; however, it is not known how these transcripts localize to the inactive X chromosome. Expression of mouse and human XIST from ectopic sites has suggested that localization to the chromosome from which the gene is expressed may be dependent upon either the copy number of the integrated constructs or the level of ectopic XIST expression. To further examine the behavior of XIST transgenes when expressed from ectopic sites, we introduced an XIST-containing PAC into the human male somatic cell line HT-1080. In five different transformant clones, the degree of localization and associated DNA condensation of the surrounding chromatin varied within nuclei of the same clone, as well as among different clones. Comparing the number of integrated transgenes and the levels of XIST expression revealed that neither factor was sufficient for a tight localization of the XIST signal. Therefore, the extent of expression and localization of XIST transcripts from ectopic transgenes is likely dependent upon many interacting factors, including the number of integrated transgenes, the level of XIST expression, and the site of integration. PMID- 12900551 TI - Functional analysis of the highly conserved exon IV of XIST RNA. AB - X inactivation is effected by a large CIS-acting RNA molecule termed the X inactive specific transcript (XIST). Exon IV of XIST RNA is highly conserved at the primary sequence level and is predicted to form a stable stem-loop structure. These features suggest that it is important for XIST RNA function. We have used homologous recombination to delete exon IV of the mouse XIST gene. Surprisingly we found no detectable effects on X inactivation. Heterozygous female animals show normal random X inactivation and transcripts from the mutant allele were seen to localise IN CIS over the length of the inactive X chromosome. There was however a reduced steady state level of mutant relative to wild type XIST RNA. This effect was not attributable to decreased stability, suggesting that the deletion affects transcription or processing of XIST RNA. PMID- 12900552 TI - Chromatin reprogramming of male somatic cell-derived XIST and TSIX in ES hybrid cells. AB - In mammalian somatic cells, the X chromosome is active in XY males, whereas one X chromosome is inactivated in XX females. On the active male X chromosome, the XIST and TSIX genes are transcribed in undifferentiated cells of pre-implantation embryos (undifferentiated state) and then down-regulated upon cell differentiation (differentiated state). To explore the epigenetic mechanism involved in the on-off switching of XIST and TSIX transcription in the active X chromosome, male somatic cells were hybridized with male embryonic stem (ES) cells. Fluorescence in situ hybridization analysis revealed that the XIST gene derived from somatic cells was derepressed, as shown by the advent of two pinpoint signals. This was confirmed by strand-specific RT-PCR of XIST and TSIX genes. To analyze changes in chromatin structure in the promoter regions of XIST and TSIX derived from somatic cells, histone tail modifications were studied by chromatin immunoprecipitation analysis. Histones H3 and H4, which were hypoacetylated in the somatic cells, were hyperacetylated in the hybrid cells, and histone H3 lysine 4, which was hypomethylated in the somatic cells, was hypermethylated in the hybrid cells, indicating that the reactivation of XIST and TSIX was linked with chromatin modifications. In the telomeric region of DXPAS34, acetylation of histones H3 and H4 was dependent on reactivation of XIST and TSIX, whereas histone H3 lysine 4 was constitutively methylated independent of the transcriptional activity of those genes. We propose that the chromatin reprogramming is linked with the resetting of the memory found in the process of choosing an active X chromosome. PMID- 12900553 TI - Effect of TSIX disruption on XIST expression in male ES cells. AB - XIST and its antisense partner, TSIX, encode non-coding RNAs and play key roles in X chromosome inactivation. Targeted disruption of TSIX causes ectopic expression of XIST in the extraembryonic tissues upon maternal transmission, which subsequently results in embryonic lethality due to inactivation of both X chromosomes in females and a single X chromosome in males. TSIX, therefore, plays a crucial role in maintaining the silenced state of XIST in CIS and regulates the imprinted X inactivation in the extraembryonic tissues. In this study, we examined the effect of TSIX disruption on XIST expression in the embryonic lineage using embryonic stem (ES) cells as a model system. Upon differentiation, XIST is ectopically activated in a subset of the nuclei of male ES cells harboring the TSIX-deficient X chromosome. Such ectopic expression, however, eventually ceased during prolonged culture. It is likely that surveillance by the X chromosome counting mechanism somehow shuts off the ectopic expression of XIST before inactivation of the X chromosome. PMID- 12900555 TI - The evolution of sex chromosomes. AB - Mammalian sex chromosomes appear, behave and function differently than the autosomes, passing on their genes in a unique sex-linked manner. The publishing of Ohno's hypothesis provided a framework for discussion of sex chromosome evolution, allowing it to be developed and challenged numerous times. In this report we discuss the pressures that drove the evolution of sex and the mechanisms by which it occurred. We concentrate on how the sex chromosomes evolved in mammals, discussing the various hypotheses proposed and the evidence supporting them. PMID- 12900554 TI - Sex chromosome elimination, X chromosome inactivation and reactivation in the southern brown bandicoot Isoodon obesulus (Marsupialia: Peramelidae). AB - Cytogenetic studies have shown that bandicoots (family Peramelidae) eliminate one X chromosome in females and the Y chromosome in males from some somatic tissues at different stages during development. The discovery of a polymorphism for X linked phosphoglycerate kinase (PGK-1) in a population of Isoodon obesulus from Mount Gambier, South Australia, has allowed us to answer a number of long standing questions relating to the parental source of the eliminated X chromosome, X chromosome inactivation and reactivation in somatic and germ cells of female bandicoots. We have found no evidence of paternal PGK-1 allele expression in a wide range of somatic tissues and cell types from known female heterozygotes. We conclude that paternal X chromosome inactivation occurs in bandicoots as in other marsupial groups and that it is the paternally derived X chromosome that is eliminated from some cell types of females. The absence of PGK 1 paternal activity in somatic cells allowed us to examine the state of X chromosome activity in germ cells. Electrophoresis of germ cells from different aged pouch young heterozygotes showed only maternal allele expression in oogonia whereas an additional paternally derived band was observed in pre-dictyate oocytes. We conclude that reactivation of the inactive X chromosome occurs around the onset of meiosis in female bandicoots. As in other mammals, late replication is a common feature of the Y chromosome in male and the inactive X chromosome in female bandicoots. The basis of sex chromosome loss is still not known; however later timing of DNA synthesis is involved. Our finding that the paternally derived X chromosome is eliminated in females suggests that late DNA replication may provide the imprint for paternal X inactivation and the elimination of sex chromosomes in bandicoots. PMID- 12900556 TI - Evolution of the human X--a smart and sexy chromosome that controls speciation and development. AB - In humans, as in other mammals, sex is determined by an XX female/XY male chromosome system. Most attention has focused on the small, degenerate Y chromosome, which bears the male-dominant gene SRY. The X, in contrast, has been considered a well-behaved and immaculately conserved element that has hardly changed since the pre-mammal days when it was just another autosome pair. However, the X, uniquely in the genome, is present in two copies in females and only one in males. This has had dire consequences genetically on the evolution of its activity--and now it appears, on its gene content and/or the function of its genes. Here we will discuss the origin of the human X, and the evolution of dosage compensation and gene content, in the light of recent demonstrations that particular functions in sex and reproduction and cognition have accumulated on it. PMID- 12900557 TI - The Azoospermia region AZFa: an evolutionar y view. AB - Compared to other regions on the human Y chromosome, the genomic segment encompassing the functionally defined AZFa locus has undergone higher X-Y sequence divergence, which is detectable by fluorescence in-situ hybridisation. This allows an evolutionary definition of an interval enclosing AZFa with a size of about 1.1 Mb. The region includes the genes USP9Y, DBY and UTY and is limited by evolutionary breakpoints within the PAC clones 41L06 and 46M11. These breakpoints restrict an area of possible male specific evolution that may have resulted in the acquisition of male specific functions, including a role in spermatogenesis. PMID- 12900558 TI - Speculations on the role of natural antisense transcripts in mammalian X chromosome evolution. AB - Recent comprehensive transcriptome analyses in mice have revealed tremendous numbers of natural antisense transcripts in a hitherto ignored category of genes in eukaryotes. We discuss the possible biological roles of these transcripts and their relationships with mammalian sex chromosome evolution. Of 60,770 full length cDNA sequences, as many as 2,500 pairs of sense-antisense transcripts (SATs) with the potential to form RNA duplex via their complementary sequences have been identified. This high number of antisense transcripts indicates their generic roles in gene expression regulation. These SATs are almost evenly distributed along the chromosomes, with the exception of the X chromosome. The rate of occurrence of SATs on the X chromosome is one-third to one-half that on the autosomes, and this under-representation must be related to a property intrinsic to the X chromosome. Here we hypothesize that monoallelically expressed antisense RNA regulates its sense partner, but that this regulatory system cannot operate on the mammalian X chromosome, as the mammalian X chromosome is effectively in a hemizygous state in both sexes. Loss of such regulation may be involved in the evolution of the X chromosome itself. PMID- 12900559 TI - Early origins of the X and Y chromosomes: lessons from tilapia. AB - Differentiated sex chromosome pairs in diverse species display certain common characteristics, normally comprising one largely heterochromatic genetically inactive chromosome and one euchromatic genetically active chromosome (e.g. the mammalian Y and X respectively). It is widely accepted that dimorphic sex chromosomes evolved from homologous pairs of autosomes. Although the exact mechanisms through which the pair diverged are not fully understood, an initial suppression of recombination in the sex-determining region is required by all of the major theories. Here we address the question of the mechanism by which this initial suppression of recombination occurs. Our model postulates that the stochastic, de novo accumulation of heterochromatin in the sex determining region can delay pairing of the sex chromosomes in meiosis, resulting in a decrease in recombination. Data to support this model is presented from the cichlid fish, Oreochromis niloticus. Although such a decrease would in most circumstances be evolutionarily disadvantageous, if the region concerned included the major sex determining gene and other gene(s) with sex-specific functions, then this would be selectively advantageous and could trigger the process(es) which, ultimately, lead to the differentiation of the sex chromosomes. PMID- 12900560 TI - Chromosome evolution in fish: sex chromosome variability in Eigenmannia virescens (Gymnotiformes: Sternopygidae). AB - New data are presented on the sex chromosomes of the fish species Eigenmannia virescens (Gymnotiformes, Sternopygidae). A new finding, involving the occurrence of ZZ/ZW sex chromosomes, is described in specimens sampled from the Sao Francisco and Amazon river basins in Brazil. All individuals had a chromosome number of 2n = 38. The homologs of the sex chromosome pair from the Sao Francisco river basin sample differed only in their morphology, while those from the Amazonian sample differed both in morphology and heterochromatin pattern. A possible model for the evolution of the sex chromosomes in E. virescens is proposed, including data from populations from the Parana (Brazil) river basin, in which male heterogamety has already been described. The occurrence of different sex chromosome systems in species and populations of the neotropical freshwater fish fauna is discussed. PMID- 12900561 TI - Sex determination and sex chromosome evolution in the medaka, Oryzias latipes, and the platyfish, Xiphophorus maculatus. AB - The first vertebrate master sex-determining gene different from Sry has been very recently discovered in a small aquarium fish, the medaka (Oryzias latipes). In this fish, the X and Y chromosomes apparently differ only by a 250-kb Y-specific region containing dmrt1bY (also called DMY and dmrt1Y), a male-specific copy of the autosomal gene dmrt1. Dmrt1 is a putative transcription factor probably involved in testis formation in different vertebrate lineages. Dmrt1bY is the only gene having escaped the drastic process of degeneration that devastated the small Y-specific region of the medaka. Mutations leading to truncation or lower expression of dmrt1bY result in XY male-to-female sex reversal. Hence, both genetic and functional evidences converge in making dmrt1bY an outstanding candidate for the function of a master sex-determining gene in fish. Nevertheless, dmrt1bY could not be detected in certain other Oryzias species or in more divergent fishes. Phylogenetic analysis revealed that the duplication of the autosomal dmrt1 that formed dmrt1bY is young in evolutionary terms. Hence, dmrt1bY is not the universal master sex-determining gene in fish. Because the classical fish models, such as zebrafish and pufferfish, are not very adequate to study the basis of genetic sex determination, alternative models, such as the platyfish (Xiphophorus maculatus), are re-emerging. In this fish, which is a well suited laboratory organism, gene loci involved in pigmentation, melanoma formation, and sexual maturity have been mapped close to the master sex determining gene. Interestingly, the platyfish can harbor three different sex chromosomes (W, X, and Y) in certain natural populations. Bacterial artificial chromosome contigs covering the sex-determining region of the platyfish are already available, and the positional cloning of the master sex-determining gene(s) should provide new insights into sex determination and sex chromosome evolution in fish and other vertebrates. PMID- 12900562 TI - Comparative mapping of Z-orthologous genes in vertebrates: implications for the evolution of avian sex chromosomes. AB - Sex chromosomes of birds and mammals are highly differentiated and share several cytological features. However, comparative gene mapping reveals extensive conserved synteny between the chicken Z sex chromosome and human chromosome 9 but not the human X sex chromosome, implying an independent origin of avian and mammalian sex chromosomes. To better understand the evolution of the avian Z chromosome we analysed the synteny of chicken Z-linked genes in zebrafish, which is the best-mapped teleost genome so far. Existing zebrafish maps do not support the existence of an ancestral Z linkage group in the zebrafish genome, whereas mammalian X-linked genes show at least some degree of synteny conservation. This is consistent with in situ hybridisation mapping data in the freshwater pufferfish, Tetraodon nigroviridis where mammalian X-linked genes show a much higher degree of conserved synteny than human chromosome 9 or the avian Z chromosome. Collectively, these data argue in favour of a more recent evolution of the avian Z chromosome, compared with the mammalian X. PMID- 12900564 TI - Cytogenetic studies of Hynobiidae (Urodela)XVIII. A ZZ/ZW sex-determining mechanism in a hynobiid salamander species, Hynobius tokyoensis Tago. AB - The karyotype of Hynobius tokyoensis (2n = 56) was analyzed using three kinds of banding methods to determine the morphological differentiation of the sex chromosomes of this species. Salamanders and egg sacs were collected from seven localities around Tokyo, Japan. Of 28 chromosome pairs, microchromosome No. 21 was identified as a ZZ/ZW-type sex chromosome. The Z chromosome was acrocentric, whereas the W chromosome was submetacentric, with a heterochromatic, elongated short arm. Interestingly, the W chromosome is of three distinct types, W(A), W(B), and W(C), based on R-banding and Ag-NOR patterns. W(A) was detected in five populations from southern habitats, whereas W(B) and W(C) were detected in one population each from northern habitats. W(A), W(B), and W(C) were all found to carry Ag-NORs on their heterochromatic short arms. Considering the karyotypes of other species belonging to the same genus, we discuss the evolution of the sex chromosomes of H. tokyoensis. PMID- 12900563 TI - The prototype of sex chromosomes found in Korean populations of Rana rugosa. AB - The seventh largest chromosome in Japanese populations of the frog Rana rugosa morphologically evolved as a sex chromosome. The sex chromosome is XX/XY type in one geographic form and ZZ/ZW type in another. In contrast, the seventh chromosomes are still homomorphic between the sexes in the other two geographic forms: they are more subtelocentric in the Kanto form and subtelocentric in the western Japanese form. To identify a prototype of the sex chromosomes, we extended our investigation in this study to the Korean form, which is supposed to be close to the phylogenetic origin of this species. The karyotype, a sex-linked gene sequence, and mechanisms of sex determination and gonadal differentiation were all examined. In addition, phylogenetic analyses were performed based on mitochondrial gene sequences and the results of crossings between the Korean and Japanese forms. As a consequence, the more subtelocentric seventh chromosome, shared by the Korean and Japanese Kanto forms, was concluded to be the prototype of the sex chromosomes. Starting at the prototype, a whole process of morphological sex chromosome evolution was reconstructed. PMID- 12900565 TI - Mammalian embryonic development--insights from studies on the X chromosome. AB - This paper reviews our early studies on the cycle of changes of X chromosome activity in different lineages of the developing female mouse embryo. The emphasis is placed on the insights gained into key developmental processes such as the temporal and spatial aspects of developmental totipotency, the timing and cell numbers involved in the origin of the germ line, the molecular mechanisms of genetic deprogramming, reprogramming and X chromosome imprinting, and transgenerational epigenetic inheritance. When viewed in this way, it is quite remarkable to see how much was learned about mammalian development from early studies on the X chromosome. Indeed several paradigm shifts occurred as a result of these studies and these are highlighted in this review. PMID- 12900566 TI - The role of the X chromosome in mammalian extra embryonic development. AB - Accumulating evidence points to the importance of the X chromosome for trophoblast development. In rodents, the extraembryonic cell lineage differs from somatic tissues in that X chromosome inactivation is imprinted, preferentially silencing the paternal X chromosome. As a consequence, trophoblast development is extremely susceptible to deviations from normal X inactivation and is impaired in situations of increased and reduced X-linked gene dosage. Mouse mutants have also shown that maintenance of X chromosome silencing in extraembryonic tissues requires a special set of heterochromatin proteins. Moreover, the X chromosome has been implicated in causing several malformations of the placenta. The observed importance of the X chromosome for placental development can be explained by the presence of many trophoblast-expressed genes, especially in the proximal and central regions. Given that the placenta represents a postzygotic barrier to reproduction, evolutionary constraints may be responsible for the presence of placental genes on the X chromosome that are often co-expressed in brain and testis. PMID- 12900567 TI - The X chromosome and ovarian function. PMID- 12900568 TI - Fertility, sex determination, and the X chromosome. AB - Because of its function, the X chromosome has a special status in mammalian genomes, with the specific occurrence of genes that influence both female and male fertility. Long ago, the XO karyotype (Turner syndrome) was associated with infertility, proving the correlation between normal X chromosome dosage and normal female fertility. Nevertheless, the search for specific X-borne fertility genes was not completely successful and suggested, instead, that female X-linked fertility, for example, depends upon groups of X-linked genes. Conversely, X linked hyperfertility has been observed in sheep, where a mutation in BMP15 leads to a hyperfertile phenotype, but only in the heterozygous state. Many male fertility genes map to the X chromosome, consistent with a genetic model developed in the early 1990s. Ironically, NR0B1 (formerly DAX1), once presented as the paradigm of genes responsible for ovarian development and function, is probably one of these male fertility factors and is active in the maintenance of spermatogenesis. Indeed, duplications of this gene on the human X chromosome lead to XY sex reversal, as NR0B1 is able to counterbalance the effect in humans. Nevertheless, invalidation experiments in mice demonstrate the effect of this factor on male germ-cell production. PMID- 12900569 TI - The role of ZIC3 in vertebrate development. AB - ZIC3 is a C2H2 zinc finger transcription factor that is involved in early patterning of the vertebrate embryo. Human patients with mutations in the X linked ZIC3 gene have a complex developmental phenotype that includes laterality defects, congenital heart disease, and lumbosacral and anal anomalies, including neural tube defects. Similar phenotypes are found in the bent tail (BN) mouse, a spontaneous mutation that is associated with a submicroscopic deletion of the ZIC3 locus, as well as in a ZIC3 null allele generated through homologous recombination. These findings suggest that ZIC3 plays important roles during development in establishing a proper left-right axis and in midline neural patterning. This review will summarize our current understanding of the role of ZIC3 in patterning of the vertebrate embryo, based on studies in model organisms such as Xenopus and the mouse. In addition, a comparison of ZIC3 with other vertebrate ZIC family members will be provided. PMID- 12900570 TI - Z and W chromosomes of chickens: studies on their gene functions in sex determination and sex differentiation. AB - Since the discovery of SRY/SRY as a testis-determining gene on the mammalian Y chromosome in 1990, extensive studies have been carried out on the immediate target of SRY/SRY and genes functioning in the course of testis development. Comparative studies in non-mammalian vertebrates including birds have failed to find a gene equivalent to SRY/SRY, whereas they have suggested that most of the downstream factors found in mammals including SOX9 are also involved in the process of gonadal differentiation. Although a gene whose function is to trigger the cascade of gene expression toward gonadal differentiation has not been identified yet on either W or Z chromosomes of birds, a few interesting genes have been found recently on the sex chromosomes of chickens and their possible roles in sex determination or sex differentiation are being investigated. It is the purpose of this review to summarize the present knowledge of these sex chromosome-linked genes in chickens and to give perspectives and point out questions concerning the mechanisms of avian sex determination. PMID- 12900572 TI - Evidence that postnatal growth retardation in XO mice is due to haploinsufficiency for a non-PAR X gene. AB - XO Turner women, irrespective of the parental source of the X chromosome, are of short stature, and this is now thought to be largely a consequence of haploinsufficiency for the pseudoautosomal region (PAR) gene SHOX. X(p)O mice (with a paternal X) are developmentally retarded in fetal life, are underweight at birth, and show reduced weight gain in the first few weeks after birth. X(m)O mice, on the other hand, are more developmentally advanced than their XX siblings in fetal life; their postnatal growth has not hitherto been assessed. Here we show that X(m)O mice are not underweight at birth, but they nevertheless show reduced weight gain postnatally. The fact that postnatal growth is affected in X(p)O and X(m)O mice, means that this must be due to X dosage deficiency. In order to see if haploinsufficiency for a PAR gene was responsible for this growth deficit (cf SHOX deficiency in Turner women), X(m)Y*(X) females, in which the Y*(X) chromosome provides a second copy of the PAR, were compared with XX females. These X(m)Y*(X) females were also growth-retarded relative to their XX sibs, suggesting that it may be haploinsufficiency for a non-dosage-compensated X gene or genes outside the PAR that is responsible for the postnatal growth deficit in XO mice. The X genes known to escape X inactivation in the mouse have closely similar Y homologues. X(m)YSRY-negative females were therefore compared with XX females to see if the presence of the SRY-negative Y chromosome corrected the growth deficit; this proved to be the case. The postnatal growth deficit of XO mice is therefore probably due to haploinsufficiency for a non-dosage compensated X gene that has a Y homologue that provides an equivalent function in the somatic tissues of males. PMID- 12900571 TI - DMRT1 in a ratite bird: evidence for a role in sex determination and discovery of a putative regulatory element. AB - Unlike mammals, birds have a ZZ male/ZW female sex-determining system. In most birds, the Z is large and gene rich, whereas the W is small and heterochromatic, but the ancient group of ratite birds are characterized by sex chromosomes that are virtually homomorphic. Any gene differentially present on the ratite Z and W is therefore a strong candidate for a sex-determining role. We have cloned part of the candidate bird sex-determining gene DMRT1 from the emu, a ratite bird, and have shown that it is expressed during the stages of development corresponding to gonadal differentiation in the chicken. The gene maps to the distal region of the Z short arm and is absent from the large W chromosome. Because most sequences on the emu W chromosome are shared with the Z, the Z-specific location constitutes strong evidence that differential dosage of DMRT1 is involved in sex determination in all birds. The sequence of emu DMRT1 has 88% homology with chicken DMRT1 and 65% with human DMRT1. Unexpectedly, an unexpressed 270-bp region in intron 3 of emu DMRT1 showed 90% homology with a sequence in the corresponding intron of human DMRT1. This extraordinarily high conservation across 300 million years of evolution suggests an important function, perhaps involved in control of DMRT1 expression and vertebrate sex determination. PMID- 12900573 TI - The X chromosome and fragile X mental retardation. AB - Fragile X syndrome represents the most common inherited cause of mental retardation. It is caused by a stretch of CGG repeats within the fragile X gene, which increases in length as it is transmitted from generation to generation. Once the repeat exceeds a threshold length, no protein is produced, resulting in the fragile X phenotype. Both X chromosome inactivation and inactivation of the FMR1 gene are the result of methylation. X inactivation occurs earlier than inactivation of the FMR1 gene. The instability to a full mutation is dependent on the sex of the transmitting parent and occurs only from mother to child. For most X-chromosomal diseases, female carriers do not express the phenotype. A clear exception is fragile X syndrome. It is clear that more than 50% of the neurons have to express the protein to ensure a normal phenotype in females. This means that a normal phenotype in female carriers of a full mutation is accompanied by a distortion of the normal distribution of X inactivation. PMID- 12900574 TI - Clinical and molecular contributions to the understanding of X-linked mental retardation. AB - X-linked mental retardation (XLMR) was first recognized in the 1940s, long before any human genes had been mapped. It is now estimated that XLMR has a prevalence of 2.6 cases per 1,000 population, accounting for over 10% of all cases of mental retardation. It is likely that over 150 genes are associated with XLMR. Fragile X syndrome, the most common form of XLMR, has a prevalence of about 1 in 4,000 males. Clinically, XLMR exists in syndromic (mental retardation with other somatic, neurological, behavioral, or metabolic findings) and nonsyndromic (mental retardation without other distinguishing features) forms. However, recent findings have caused this distinction to become blurred as mutations in some genes have been found in both syndromic and nonsyndromic XLMR. Progress in XLMR gene identification has allowed some insight into various pathways and cellular activities involved in developing cognitive functions. The genes involve signaling pathways, transcription factors, cytoskeletal organization, cell adhesion and migration, and maintenance of the cell membrane potential. PMID- 12900576 TI - Mapping of a further locus for X-linked craniofrontonasal syndrome. AB - Craniofrontonasal syndrome is a rare dysostosis syndrome with an unusual pattern of X-linked inheritance, because males are usually not or less severely affected than females. Previously, a CFNS locus has been localised in Xp22. We report on a haplotype analysis in a German CFNS family, mapping the CFNS locus to the pericentromeric region of the X chromosome. This discrepancy can be explained by locus heterogeneity. Furthermore, random X inactivation could be demonstrated in affected females. The most plausible interpretation for this unusual pattern of X linked inheritance is metabolic interference. Consequently, we propose that the CFNS gene escapes X inactivation. PMID- 12900575 TI - The proportion of cells with functional X disomy is associated with the severity of mental retardation in mosaic ring X Turner syndrome females. AB - Turner syndrome females (45,X) do not have mental retardation (MR), whereas some mosaic ring X Turner syndrome females, with 45,X/46,X,r(X), have severe MR. The MR is believed to be caused by a failure of X chromosome inactivation (XCI) of the small ring X chromosome, which leads to functional X disomy (FXD), To explore this hypothesis, we examined the proportion of FXD cells in the peripheral blood of four ring X Turner syndrome females with various levels of MR, using two newly developed XCI assays based on DNA methylation of X-linked genes. As a result, the two patients with extremely severe MR showed complete FXD patterns, whereas the remaining two patients with relatively milder MR showed partial FXD patterns. These results indicate that the proportion of FXD cells may be associated with the severity of MR in mosaic ring X Turner syndrome females, although this association should be confirmed by examining brain cells during development. One of the cases with severe MR and a complete FXD pattern neither lacked the XIST gene nor had uniparental X isodisomy, and we discuss the mechanism of the failure of XCI in this case. PMID- 12900577 TI - Microphthalmia with linear skin defects (MLS), Aicardi, and Goltz syndromes: are they related X-linked dominant male-lethal disorders? AB - Gene identification for X-linked dominant sporadic disorders is challenging because no extended families exist that can be studied by linkage analysis. Therefore, classic positional cloning approaches are not possible, and other methods have to be used to search for candidate genes. These conditions present the next challenge for disease-gene identification of Mendelian disorders. The various issues and difficulties involved, such as male lethality, X chromosome inactivation, and analysis of phenotypic similarities among different conditions are illustrated through discussion of three X-linked developmental disorders: microphthalmia with linear skin defects (MLS) syndrome, Aicardi syndrome, and Goltz syndrome (focal dermal hypoplasia). PMID- 12900578 TI - Microphthalmia with linear skin defects syndrome (MLS): a male with a mosaic paracentric inversion of Xp. AB - The microphthalmia with linear skin defects syndrome (MLS) is an X-linked dominant disorder with male lethality. In the majority of the patients reported, the MLS syndrome is caused by segmental monosomy of the Xp22.3 region. To date, five male patients with MLS and 46,XX karyotype ("XX males") have been described. Here we report on the first male case with MLS and an XY complement. The patient showed agenesis of the corpus callosum, histiocytoid cardiomyopathy, and lactic acidosis but no microphthalmia, and carried a mosaic subtle inversion of the short arm of the X chromosome in 15% of his peripheral blood lymphocytes, 46,Y,inv(X)(p22.13 approximately 22.2p22.32 approximately 22.33)[49]/46,XY[271]. By fluorescence IN SITU hybridization (FISH), we showed that YAC 225H10 spans the breakpoint in Xp22.3. End-sequencing and database analysis revealed a YAC insert of at least 416 kb containing the genes HCCS and AMELX, and exons 2-16 of ARHGAP6. Molecular cytogenetic data suggest that the Xp22.3 inversion breakpoint is located in intron 1 of ARHGAP6, the gene encoding the Rho GTPase activating protein 6. Future molecular studies in karyotypically normal female MLS patients to detect submicroscopic rearrangements including the ARHGAP6 gene as well as mutation screening of ARHGAP6 in patients with no obvious chromosomal rearrangements will clarify the role of this gene in MLS syndrome. PMID- 12900579 TI - X-chromosomal localization of mammalian Y-linked genes in two XO species of the Ryukyu spiny rat. AB - Ryukyu spiny rats (genus Tokudaia), which are endemic to the central part of the Nansei Shoto archipelago in Japan, have unique karyotypes with odd numbers of chromosomes and no cytologically recognizable Y chromosome. The chromosome numbers of Tokudaia osimensis from Amamioshima and of Tokudaia sp. from Tokunoshima are 2n = 25 and 2n = 45, respectively, with a putative single X chromosome. The mouse X probe hybridized to the unpaired X chromosome, except for the distal part of the short arm in a female specimen of T. osimensis and in one male and one female of Tokudaia sp. Fluorescence in situ hybridization with the Tspy (testis-specific protein, Y-encoded) gene from both male and female cells of Tokudaia sp. by PCR localized Tspy to the distal part of the long arm of the X chromosome. Another Y-related gene, Zfy, from Tokudaia sp. was also localized to the same region in both species. Although the Sry gene is absent in this species, the present results suggest that the Y-chromosome segment carrying functional Y linked genes, such as Tspy and Zfy, is translocated onto the distal part of the long arm of the X chromosome. PMID- 12900580 TI - Biallelic expression of Z-linked genes in male chickens. AB - In birds, females are heterogametic (ZW), while males are homogametic (ZZ). It has been proposed that there is no dosage compensation for the expression of Z linked genes in birds. In order to examine if the genes are inactivated on one of the two Z chromosomes, we analyzed the allelic expression of the B4GALT1 and CHD Z genes on Z chromosomes in male chickens. One base substitution was detected among 15 chicken breeds and lines examined for each gene, and cross mating was made between the breeds or lines with polymorphism. cDNAs were synthesized from cultured cell colonies each derived from a single cell of an F1 male embryo. The allelic expression of the B4GALT1 gene was examined by restriction fragment length polymorphism analysis of the PCR products digested with RSAI, and that of the CHD-Z gene by the single nucleotide primer extension (SNuPE) method. Both of the genes displayed biallelic expression, suggesting that these Z-linked genes were not subject to inactivation in male chickens. Comparison between expression levels in males and females by real-time quantitative PCR suggested that expression was compensated for the CHD-Z gene but not for the B4GALT1 gene. PMID- 12900581 TI - X chromosomes of American marsupials contain minimal amounts of euchromatin. AB - The karyotypes of four South American didelphid marsupials, representing diploid numbers of 2n = 14 and 18, have been analyzed by a variety of banding techniques. The 2n = 14 karyotypes display a high degree of homoeology, but there also exist distinct similarities between the 2n = 14 and 2n = 18 karyotypes. The interspecific differences found are due to centric fissions, pericentric inversions, and variations in the amount and composition of the constitutive heterochromatin. Contrary to the evolutionary conservation of the banding patterns in all autosomal arms, there are multiple differences in the number and chromosomal location of the nucleolus organizer regions. In species with X-linked nucleolus organizers, the 18S + 28S ribosomal RNA genes escape inactivation in female cells. Measurements on the X chromosomes of Marmosa fuscata and Micoureus demerarae unexpectedly reveal the lowest quantities of euchromatin so far determined in the X chromosomes of mammals: 1.5% and 1.8%, respectively, of their haploid female genomes. This is significantly less than the amount of euchromatin in the basic X chromosomes of other marsupials (3%) or eutherians (5%). PMID- 12900582 TI - Reorganization of the X chromosome in voles of the genus Microtus. AB - Comparative chromosomal analysis is a powerful tool in the investigation of the mechanisms of chromosomal evolution. The accuracy of the analysis depends on the availability of region-specific markers to follow the fate of the particular chromosomal region through the evolution of species. We have assigned 12 unique sequences to the euchromatic part of the vole X chromosome, which serve as reliable markers of chromosomal segments. Together with region-specific libraries and GTG banding, these markers allow us to delineate the homologous regions of the X chromosomes in five species of the genus Microtus. We found that X chromosomes of these species differ by numerous rearrangements and all rearrangements are clustered at specific breakpoints. Moreover, these breakpoints were found to colocalise with repetitive and/or duplicated DNA sequences. We suggest that clusters of repeated and/or duplicated DNA sequences have played a crucial role in the formation of rearrangement hot spots during evolution of the X chromosome in the subgenus Microtus. PMID- 12900583 TI - Chromosome banding in Amphibia. XXVI. Coexistence of homomorphic XY sex chromosomes and a derived Y-autosome translocation in Eleutherodactylus maussi (Anura, Leptodactylidae). AB - A 15-year cytogenetic survey on one population of the leaf litter frog Eleutherodactylus maussi in northern Venezuela confirmed the existence of multiple XXAA male symbol /XAA(Y) female symbol sex chromosomes which originated by a centric (Robertsonian) fusion between the original Y chromosome and an autosome. 95% of the male individuals in this population are carriers of this Y autosome fusion. In male meiosis the XAA(Y) sex chromosomes pair in the expected trivalent configuration. In the same population, 5% of the male animals still possess the original, free XY sex chromosomes. In a second population of E. maussi analyzed, all male specimens are characterized by these ancestral XY chromosomes which form normal bivalents in meiosis. E. maussi apparently represents the first vertebrate species discovered in which a derived Y-autosome fusion still coexists with the ancestral free XY sex chromosomes. The free XY sex chromosomes, as well as the multiple XA(Y) sex chromosomes are still in a very primitive (homomorphic) stage of differentiation. With no banding technique applied it is possible to distinguish the Y from the X. DNA flow cytometric measurements show that the genome of E. maussi is among the largest in the anuran family Leptodactylidae. The present study also supplies further data on differential chromosome banding and fluorescence in situ hybridization experiments in this amphibian species. PMID- 12900584 TI - The X chromosome deletion in HYP mice extends into the intergenic region but does not include the SAT gene downstream from Phex. AB - The murine Hyp mutation is a model for X-linked hypophosphatemia (XLH), the most prevalent form of inherited rickets in humans. Although mutations in the murine Phex gene and the human PHEX gene have been identified in both murine and human disorders, the extent of the Hyp deletion on the mouse X chromosome has not been delineated. In the present study we demonstrate that the Hyp deletion starts in the middle of Phex intron 15 and includes approximately 48 kb of the 3' region of the Phex gene and approximately 10 kb of intergenic sequence on the mouse X chromosome. In addition, we show that the Hyp deletion does not involve the downstream spermidine/spermine N1-acetyl transferase (Sat; formerly Ssat) gene and thus is not a contiguous gene deletion syndrome. Our data indicate that the Hyp mouse is a true homolog of XLH in humans and underscore the validity of this murine model in studies of XLH pathophysiology and for testing novel treatment modalities. PMID- 12900585 TI - A practical approach to persistent elevation of blood pressure in the hypertension clinic. AB - Patients seen with persistent elevation of office blood pressure (BP) in the hypertension clinic are common and pose a significant challenge to the hypertension specialist. These are usually patients sent to the clinic because of difficulty in achieving BP goals. In this clinical setting there is usually data available from home self measurement and ambulatory BP readings. We propose a scheme to encourage specialists to consciously consider the causes of persistent elevation of office BP. The scheme relies initially on consideration that the measured office readings are falsely elevated due to pseudohypertension, small cuff size or other equipment problems. If this is not the case then the measured office BP is truly elevated. If so, a further distinction can be made in this group into those with (1). transient office BP elevation and normal home and/or ambulatory readings and (2). elevated office and out of office readings. The first subgroup is usually called white-coat hypertension but may also be seen with transient increases in BP due to talking or other factors. The second subgroup probably is the majority of patients with persistent high office BP and reflects sustained hypertension and its own list of causes including secondary hypertension. The nature and intensity of evaluation for secondary hypertension varies depending on the clinical presentation of the patient. We recommend an explicit consideration of reasons for persistent high office BP in the hypertension clinic. PMID- 12900586 TI - Effect of standard cuff on blood pressure readings in patients with obese arms. How frequent are arms of a 'large circumference'? AB - OBJECTIVE: To measure the effect on blood pressure readings when a standard cuff is used on patients with arms of a large circumference, and to determine the frequency of arms of a large circumference. SUBJECTS: Blood pressures were taken in 120 subjects with an arm circumference greater than 33 cm. Also, the arm circumference was determined in 244 patients from a family health unit, and in 216 patients from a hypertension clinic. METHOD: A mercury sphygmomanometer and two different cuff sizes were used in a random sequence; therefore, 60 patients' blood pressure were first measured with a large cuff, followed by a standard cuff; the opposite sequence was then applied for another 60 patients. With the obtained values and using a regression analysis, the difference in blood pressure overestimation was calculated. Arm circumference measurement percentages were used to determine the frequency of arms of a large circumference. RESULTS: Both systolic and diastolic blood pressures were significantly greater when the standard cuff was used. For every 5 cm increase in arm circumference, starting at 35 cm, a 2-5 mmHg increase in systolic blood pressure, and a 1-3 mmHg increase in diastolic blood pressure was observed. The prevalence of arms with a large circumference in the family medicine unit and hypertension clinic was 42% and 41.8%, respectively. CONCLUSIONS: There is an overestimation of blood pressure when a standard cuff is used in obese subjects. The high prevalence of these individuals in our environment, both in the hypertensive and normotensive population, makes it necessary to have on hand different sizes of cuffs for taking blood pressure in order to avoid incorrect decisions. PMID- 12900587 TI - Ambulatory blood pressure correlates with renal volume and number of renal cysts in children with autosomal dominant polycystic kidney disease. AB - OBJECTIVE: In adult patients with autosomal dominant polycystic kidney disease (ADPKD) renal volume was found to be significantly greater in hypertensive compared to normotensive patients. The purpose of this study was to find out if blood pressure (BP) is related to renal size also in children with ADPKD, for example, in an early stage of the disease. METHOD AND RESULTS: Sixty-two children with ADPKD and normal renal function (mean age 12.3+/-4.3 years) were examined by renal ultrasound and ambulatory BP monitoring (ABPM). Twenty-two children were hypertensive and 40 normotensive. Mean renal volume was significantly greater in hypertensive than in normotensive children (2.7+/-2.3 SDS versus 1.2+/-2.5 SDS, P<0.01) despite similar anthropometric data and renal function. Similarly the mean number of cysts was significantly higher in hypertensive patients than in normotensive (35+/-15 cysts versus 23+/-14 cysts, P<0.01). Renal volume correlated with daytime as well as with night-time systolic and diastolic BP (r=0.41-0.47, P<0.01). Correlations with renal length and the number of renal cysts were somewhat less (r=0.29-0.43, P<0.05 and 0.01, respectively). CONCLUSIONS: This study revealed a significant relationship between renal volume, renal length and number of renal cysts and BP. It is suggested that children with ADPKD should regularly be checked for BP changes by ABPM, especially those who show increased renal size or a high number of renal cysts on ultrasound. All these children are at high risk for development of hypertension. PMID- 12900588 TI - Comparison of telmisartan versus losartan: meta-analysis of titration-to-response studies. AB - OBJECTIVES: To compare the ability of telmisartan and losartan to reduce mean diastolic blood pressure (DBP) during the last 6 h of the 24-h dosing interval in a prospectively planned meta-analysis of ambulatory blood pressure monitoring (ABPM) data from two independent studies. METHODS: Data were from two independent randomized, double-blind, double-dummy, titration-to-response studies conducted in patients with mild-to-moderate hypertension (seated cuff DBP 95-109 mmHg, 24-h mean ambulatory DBP >or=85 mmHg). After a 4-week placebo run-in period, patients received once-daily telmisartan 40 mg or losartan 50 mg, with up-titration after 4 weeks to telmisartan 80 mg or losartan 100 mg, respectively, if seated trough cuff DBP >or=90 mmHg. Blood pressures were recorded using ABPM immediately before randomization and after 8 weeks of active treatment. In addition, seated trough cuff blood pressures were measured at baseline and after 4 and 8 weeks of active treatment. RESULTS: Titration to the higher dose was required in 60.1% of telmisartan patients and 69.5% of losartan patients (P=0.01). Reductions from baseline in the last 6 h mean ambulatory DBP with telmisartan and losartan were 6.6+/-0.4 and 5.1+/-0.4 mmHg, respectively (P<0.01, adjusted for baseline and study); the effects were homogeneous across the two studies. During the last 6 h of the 24-h dosing interval, telmisartan produced greater reductions in each of the observed hourly mean ambulatory DBP values. Telmisartan-induced reductions were also greater for the majority of the observed hourly mean ambulatory DBP values over the entire 24-h dosing interval. Reductions from baseline in the last 6 h adjusted mean ambulatory systolic blood pressure (SBP) for telmisartan and losartan were 9.9+/-0.6 and 7.8+/-0.6 mmHg, respectively (P=0.01). The 24-h profiles of ambulatory SBP hourly mean reductions were similar to those for DBP. Both telmisartan and losartan were found to be safe and well tolerated. CONCLUSIONS: Telmisartan 40/80 mg is superior to losartan 50/100 mg in controlling DBP and SBP during the last 6 h of the 24-h dosing interval. PMID- 12900589 TI - Effect of doxazosin GITS on 24-hour blood pressure profile in patients with stage 1 to stage 2 primary hypertension. AB - OBJECTIVE: To investigate the effect of the doxazosin gastrointestinal therapeutic system (GITS) on the 24 h blood pressure (BP) profile by ambulatory blood pressure measurements (ABPM) in patients with stage 1 to stage 2 primary hypertension. METHODS AND RESULTS: Seventeen hypertensive patients-either untreated or after a two-week run-in/washout period-underwent office and ABPM monitoring before and six weeks after an open-label once-daily morning dose of 4 mg of doxazosin GITS, an alpha(1)-adrenoceptor antagonist. Fourteen patients responded; three did not. Data analyses refers to the responders: linear analysis demonstrated statistically significant reductions from baseline in daytime, night time, and total 24 h means for systolic BP (SBP) (7-10 mmHg) and diastolic BP (DBP) (5-10 mmHg) after treatment, with no statistically significant change in heart rate (HR). Rhythm analysis demonstrated statistically significant reductions from baseline in mean mesor (8 mmHg), maximum (6 mmHg) and minimum (10 mmHg) values in SBP, and in mean mesor (5 mmHg), maximum (7 mmHg) and minimum (5 mmHg) values in DBP. Circadian rhythm parameters in BP and HR were not significantly altered by treatment. Treatment with doxazosin GITS was well tolerated. CONCLUSIONS: A single morning dose of doxazosin GITS at 4 mg significantly reduced ambulatory SBP and DBP throughout a 24 h period while preserving a normal 24 h BP and HR rhythm profile in stage 1 to stage 2 hypertensives. PMID- 12900590 TI - Evaluation of two devices for self-measurement of blood pressure according to the international protocol: the Omron M5-I and the Omron 705IT. AB - BACKGROUND: Two devices for self-measurement of blood pressure at the brachial artery-the Omron M5-I and the Omron 705IT-were evaluated according to the international protocol of the European Society of Hypertension. DESIGN: The international validation protocol is divided into two phases: the first phase is performed on 15 selected subjects and if the device passes this phase, 18 supplementary subjects are included making a total number of 33 subjects on which the final validation is performed. METHODS: For each subject, four blood pressure (BP) measurements were performed simultaneously by two trained observers using mercury sphygmomanometers alternately with three measurements by the tested device. The difference between the BP value given by the device and that obtained by the two observers (mean of the two observers) was calculated for each measure. The 99 differences were classified into categories ( 0.05). There was also no significant difference in the complication rate of both of the Childrens Hospital Los Angeles groups compared with the Royal Children's Hospital group (p > 0.05). This study indicates that cleft lip repair performed in an outpatient setting may be a safe alternative to the inpatient operation. Certain preexisting medical conditions, however, may dictate the need for inpatient hospitalization after repair. PMID- 12900595 TI - Secondary rhinoplasty using flying-bird and vestibular tornado incisions for unilateral cleft lip patients. AB - To correct the nasal deformity in cleft lip patients, a new procedure of open rhinoplasty using a "flying-bird" incision in the nostril tip with a vestibule "tornado"-shaped incision in the cleft side is presented. The newly designed vestibular incision produces effective vestibular advancement with the freed lower lateral cartilage. The flying-bird incision makes it possible to produce a suitable nostril tip appearance with symmetrical external nostril vestibules. If the vestibular defect after flap advancement is wide, a full-thickness skin graft is used to give priority for making a good external nostril shape. This procedure is useful for most cleft lip noses, particularly in cases of moderate to severe deformity. PMID- 12900596 TI - Blind canalicular marsupialization in complete punctal absence as part of a systematic approach for classification and treatment of lacrimal system obstructions. AB - The procedure of choice for epiphora caused by bipunctal and canalicular obstruction is conjunctivodacryocystorhinostomy. Despite its high success rate, it may result in multiple complications, such as extrusion, migration, and secondary obstruction. The author describes a simple alternative procedure to conjunctivodacryocystorhinostomy for patients with epiphora caused by bipunctal and proximal canaliculus complete occlusion and a systematic approach to treat lacrimal system obstructions. Ten instances of bipunctal and proximal canaliculus absence in five consecutive patients, caused in four patients by ocular surface disorders (topical drug toxicity, herpetic keratoconjunctivitis, and trachoma), were treated by blunt dissection of the presumed lower punctal site under a surgical microscope. The punctal site was determined by several landmarks, the peaked medial lid margin, a dimple at that site, or an area of relative avascularity. The canaliculus was exposed and expanded to create a pocket. After the procedure, the lacrimal drainage system was found patent in nine of the 10 procedures. After one additional procedure, irrigation of the lacrimal drainage system revealed a nasolacrimal duct obstruction that was treated with dacryocystorhinostomy and silicone tube insertion. After these procedures, an objective resolution of the epiphora was noted in all patients. Epiphora resulting from lack of punctal and proximal canaliculus caused by ocular surface diseases may be treated with blind exposure and marsupialization of the proximal canaliculus instead of conjunctivodacryocystorhinostomy. If, in addition, the nasolacrimal duct is obstructed, a dacryocystorhinostomy may be performed. If this proposed procedure fails, the patient can still undergo conjunctivodacryocystorhinostomy or other procedures. The procedure may be part of a systematic approach to treat lacrimal drainage obstructions that is based on an association between the location and the cause of the obstruction. PMID- 12900597 TI - The free deltoid flap: microscopic anatomy studies and clinical application to oral cavity reconstruction. AB - The deltoid free flap is a fasciocutaneous flap that should be thin, hairless, of an adequate size, and capable of sensory reinnervation. Because of its excellent color-matching and texture-matching characteristics, it has recently been widely used for the reconstruction of soft-tissue defects during oral and maxillofacial surgery. Furthermore, a characteristic of oral and maxillofacial soft-tissue defects is that they are not large; therefore, flap size will be small, allowing the donor site to be directly closed. Because of natural variation in parts of the anatomy, there has sometimes been great difficulty in clinical application. The authors decided to study this by performing anatomical studies of the deltoid region on 21 cadavers. The result indicates that the pedicle of the deltoid free flap penetrates the "quadrangular space" in 90 percent of cases but passes and does not penetrate the quadrangular space in the remaining cases. The authors also confirmed that the skin has a vascular network comprising five layers and, furthermore, that the vascular network of the deep fascia is dense. The authors also report six cases of its clinical use complicated by anatomic variation and local infection in which the deltoid flap showed a completely successful outcome. PMID- 12900598 TI - Algorithm of hair restoration surgery in children. AB - Hair is an inseparable element of external appearance of every human being. Although various fashion trends come and go, the lack of hair is for many a major aesthetic and psychological problem. Even if men's balding can be accepted as a natural phenomenon, hair loss in children is considered to be a condition demanding correction. During an 18-year period, 8440 hair restoration operations were performed at the Hair Clinic Poznan, in Poznan, Poland. Most patients were men treated for androgenic alopecia. Among the patients were 57 children in whom hair loss resulted from hereditary factors, perinatal traumas, radiotherapy, and mechanical, thermal, and chemical damage. Methods of restoration were adjusted to type of hair loss, patient age, and ability to cooperate with the surgeon. In cases of single massive scars, skin flap correction was usually used. The flaps were prepared with the use of expanders. In cases of numerous scattered defects or considerable thinning of the scalp, the method of choice was hair transplantation. The "four-hand stick-and-place" technique developed by the authors enabled the surgeon to quickly and precisely carry out the procedure. Application of varied surgery techniques in scalp reconstruction procedures in children gave very good aesthetic results with a minimal complication rate. PMID- 12900599 TI - Hair-bearing submental artery island flap for reconstruction of mustache and beard. AB - Loss of mustache and beard in the adult male caused by severe burn, trauma, or tumor resection may cause cosmetic and psychological problems for these patients. Reconstruction of the elements of the face presents difficult and often daunting problems for plastic surgeons. The tissue that will be used for this purpose should have the same characteristics as the facial area, consisting of thin, pliable, hair-bearing tissue with a good color match. There is a very limited amount of donor area that has these characteristics. A hair-bearing submental island flap was used successfully for mustache and beard reconstruction in 11 male patients during the last 5 years. The scar was on the mentum in four cases, right cheek in two cases, right half of the upper lip in two cases, left cheek in one case, left half of the upper lip in one case, and both sides of the upper lip in one case. The submental island flap is supplied by the submental artery, a branch of the facial artery. The maximum flap size was 13 x 6 cm and the minimum size was 6 x 3 cm (average, 10 x 4 cm) in this series. Direct closure was achieved at all donor sites. Patients were followed up for 6 months to 5 years. No major complication was noted other than one case of temporary palsy of the marginal mandibular branch of the facial nerve. The mean postoperative stay was 7 days. Color and texture match were good. Hair growth on the flap was normal, and characteristics of the hair were the same as the intact side of the face in all patients. The submental island flap is safe, rapid, and simple to raise and leaves a well-hidden donor-site scar. The authors believe that the submental artery island flap surpasses the other flaps in reconstruction of the mustache and beard in male patients. Application of the technique and results are discussed in this article. PMID- 12900600 TI - Comparison of donor-site healing under Xeroform and Jelonet dressings: unexpected findings. AB - Split-thickness skin grafts remain central to the strategy of burn wound treatment. The dressing used to cover the donor wound site has a significant effect on healing parameters. The purpose of this study was to compare split thickness skin graft donor site reepithelialization under Xeroform and Jelonet dressings. A dermatome was used to cut two consecutive strips of skin from 25 paired donor sites on the thigh, calf, or back of 19 participants. Standardization of the harvest method was achieved by using the same surgeon to harvest the compared skin graft strips, with attention to consistency of dermatome skin-thickness setting, downward pressure, and angle of dermatome approach. A strip of Xeroform or Jelonet was applied to one of each pair of wounds. Epidermal and dermal thickness was measured from biopsy specimens cut at the midpoint of each split-thickness graft strip. The day of final dressing separation was declared the day of complete donor reepithelialization (healing). The mean healing time for Xeroform and Jelonet was 10.4 +/- 2.6 days (n = 25) and 10.6 +/- 2.8 days (n = 25) (p = 0.76) at sites cut to a mean depth of 0.23 +/- 0.08 mm and 0.23 +/- 0.09 mm (p = 0.89), respectively. There was no correlation between graft thickness and healing time for sites dressed with Xeroform (r = 0.17) or Jelonet (r = 0.02). Donors sites reharvested 10 to 21 days after a prior harvest healed an average of 3.1 days earlier than virgin sites (8.4 +/- 1.6 versus 11.5 +/- 2.6 days, p < 0.001), although reharvested grafts were on average 0.05 mm thicker (p = 0.10). The mean thickness of reepithelialized donor-site epidermis (0.13 +/- 0.04 mm, n = 30) was found to be twice the thickness of virgin epidermis from the same sites (0.06 +/- 0.02 mm, n = 38, p < 0.001). Thirty-six grafts harvested with dermatomes set to cut 8/1000 inch (0.20 mm) deep ranged from 0.12 to 0.42 mm thick, with only eight of these grafts measuring within +/-10 percent of the desired thickness setting. Before donor dressing separation, Xeroform and Jelonet dressings were judged to be more comfortable by nine patients and one patient, respectively, whereas no difference was detected by six patients. The authors now use Xeroform as the preferred donor dressing. PMID- 12900601 TI - Aesthetic subunits of the breast. AB - Surgery for breast cancer has traditionally addressed the breast as if it were a geometric circle with associated quadrants. Cosmetic reconstruction should not follow geometric patterns but should emphasize perceived contour and normal clothing lines. Similar to nasal reconstruction, a subunit principle in breast reconstruction planning may significantly improve the appearance of the result. To better identify the most aesthetic subunits for breast reconstruction, 10 years of autogenous reconstruction in 264 patients was reviewed. Various patterns of breast subunits were identified. The more favorable subunits of the breast in terms of postoperative appearance and camouflage of scars included the nipple, areola, and expanded areola subunits. For larger skin defects, the best subunits were the inferolateral, lower half, and a total breast subunits. Dividing the breast into reconstructive subunits that are to be replaced as a whole rather than as a patch gives superior results. Photographed examples of aesthetic subunits illustrate the placement of scars along natural lines that maximize the advantages of camouflage afforded by clothing. PMID- 12900602 TI - Alternative venous outflow vessels in microvascular breast reconstruction. AB - The lack of adequate recipient vessels often complicates microvascular breast reconstruction in patients who have previously undergone mastectomy and irradiation. In addition, significant size mismatch, particularly in the outflow veins, is an important contributor to vessel thrombosis and flap failure. The purpose of this study was to review the authors' experience with alternative venous outflow vessels for microvascular breast reconstruction. In a retrospective analysis of 1278 microvascular breast reconstructions performed over a 10-year period, the authors identified all patients in whom the external jugular or cephalic veins were used as the outflow vessels. Patient demographics, flap choice, the reasons for the use of alternative venous drainage vessels, and the incidence of microsurgical complications were analyzed. The external jugular was used in 23 flaps performed in procedures with 22 patients. The superior gluteal and transverse rectus abdominis musculocutaneous (TRAM) flaps were used in the majority of the cases in which the external jugular vein was used (72 percent gluteal, 20 percent TRAM flap). The need for alternative venous outflow vessels was usually due to a significant vessel size mismatch between the superior gluteal and internal mammary veins (74 percent). For three of the external jugular vein flaps (13 percent), the vein was used for salvage after the primary draining vein thrombosed, and two of three flaps in these cases were eventually salvaged. In three patients, the external jugular vein thrombosed, resulting in two flap losses, while the third was salvaged using the cephalic vein. A total of two flaps were lost in the external jugular vein group. The cephalic vein was used in 11 flaps (TRAM, 64.3 percent; superior gluteal, 35.7 percent) performed in 11 patients. In five patients (54.5 percent), the cephalic vein was used to salvage a flap after the primary draining vein thrombosed; the procedure was successful in four cases. In three patients, the cephalic vein thrombosed, resulting in two flap losses. One patient suffered a thrombosis after the cephalic vein was used to salvage a flap in which the external jugular vein was initially used, leading to flap loss, while a second patient experienced cephalic vein thrombosis on postoperative day 7 while carrying a heavy package. There was only one minor complication attributable to the harvest of the external jugular or cephalic vein (small neck hematoma that was aspirated), and the resultant scars were excellent. The external jugular and cephalic veins are important ancillary veins available for microvascular breast reconstruction. The dissection of these vessels is straightforward, and their use is well tolerated and highly successful. PMID- 12900603 TI - The correction of capsular contracture by conversion to "dual-plane" positioning: technique and outcomes. AB - Little has been published regarding the treatment of patients with long established capsular contracture after previous submuscular or subglandular breast augmentation. This study reviews 7 years of experience in treating established capsular contracture after augmentation mammaplasty by relocating implants to the "dual-plane" or partly subpectoral position. A retrospective chart review was performed on all patients who were treated for capsular contracture using this technique between 1993 and 1999. Data collected included the date of the original augmentation, the original implant location, date of revision and type of implant used, length of follow-up, outcome, and any ensuing complications. Different surgical techniques were used, depending on whether the prior implant was located in a subglandular or submuscular plane. All patients had revisions such that their implants were relocated to a dual plane, with the superior two thirds or so of the implant located beneath the pectoralis major muscle and the inferior one third located subglandularly. Of 85 patients reviewed, 54 had their original implants in a submuscular position and 31 had their initial augmentation in a subglandular position. Of the 54 patients whose implants were initially submuscular, 23 patients (43 percent) had silicone gel implants, 15 patients (28 percent) had double-lumen implants, and the remaining 16 patients (30 percent) had saline implants. Of the 31 patients whose implants were initially subglandular, 20 patients (65 percent) had silicone gel implants, three patients (10 percent) had double-lumen implants, and the remaining eight patients (26 percent) had saline implants. Fifty-one patients (60 percent) had replacement with saline implants (37 smooth saline, 14 textured saline), whereas 34 (40 percent) had silicone gel implants (seven smooth gel, 27 textured gel). The average time from previous augmentation to revision was 9 years 9 months. The average follow-up time after conversion to the dual-plane position was 11.5 months. Only three of 85 patients required reoperation for complications, all of which involved some degree of implant malposition. Of patients converted to the dual plane, 98 percent were free of capsular contracture and were Baker class I at follow-up, whereas 2 percent were judged as Baker class II. There were no Baker level III or IV contractures at follow-up. The dual-plane method of breast augmentation has proved to be an effective technique for correcting established capsular contracture after previous augmentation mammaplasty. This technique appears to be effective when performed with either silicone or saline-filled implants. PMID- 12900604 TI - Infectious complications following breast reconstruction with expanders and implants. AB - The incidence of infection following breast reconstruction with expanders and implants ranges from 1 to 24 percent. Numerous factors associated with infection have been described; however, a one-variable at time setting and multifactorial analysis have not been performed. The purpose of this study was to analyze a set of factors that may predispose women to infection of the expander or implant. Between 1997 and 2000, a total of 168 implant reconstructions were performed in 130 women at a single institution. The mean age for all women was 48.2 years (range, 25 to 77 years). The factors that were analyzed included axillary lymph node dissection, chemotherapy, radiation therapy, tumor stage, timing of implant insertion, number of sides (unilateral versus bilateral), tobacco use, and presence or absence of diabetes mellitus. Statistical analysis was performed with stepwise logistic regression. Mean time to follow-up for all patients was 29 months (range, 12 to 47 months). Infectious complications occurred in 10 women (7.7 percent) and in 10 expanders or implants (5.9 percent). Infected implants were removed an average of 116 days following insertion (range, 14 to 333 days). Cultured bacteria included Staphylococcus aureus and Serratia marcescens. A significant association (p < 0.04) was detected between implant infection and radiation therapy. The chance for implant infection was 4.88 times greater for implants that were exposed to radiation therapy compared with those that were not. In addition, there was suggestive (p < 0.09) evidence that the chance of implant infection following lymph node dissection was 6.29 times higher than when no lymph nodes were removed. No significant association between implant infection and age, diabetes, tobacco use, tumor stage, timing of implant insertion, or chemotherapy was found. PMID- 12900605 TI - Abdominal wall partitioning (the accordion effect) for reconstruction of major defects: a retrospective review of 10 patients. AB - Ten patients underwent abdominal wall reconstruction using the technique of abdominal wall partitioning. All defects were closed in the midline by approximating fascia to fascia with the assistance of a general surgeon. One patient had skin grafted small bowel. Five patients had chronically infected mesh and previous failed attempts at repair. Four patients had large ventral hernias following gastric reduction operations and massive weight loss. No defect in any dimension was less than 20 cm. All patients had secure abdominal wall repair by reconstruction of a midline anchor for the abdominal wall musculature. One patient was lost to follow-up after 3 weeks. The average follow-up time for the remaining nine patients was 18.6 months (range, 6 months to 4.7 years). One patient required readmission to the hospital for management of a limited area of skin necrosis. Two patients had minor wound infections, and three patients had subcutaneous seromas, all of which were managed on an outpatient basis. One patient developed a 2 x 2-cm subxiphoid hernia recurrence. Technical details include subcutaneous undermining of the abdominal skin to the anterior axillary lines bilaterally, mobilization of the viscera to expose the white lines of Toldt bilaterally, and parallel, parasagittal, staggered releases of the transversalis fascia, transversalis muscle, external oblique fascia, external oblique muscle, and rectus fascia. These multiple releases allow expansion and translation of the abdominal wall by an accordion-like effect. This accordion-like effect allows closure of abdominal wall defects that are substantially larger than what can be closed with current techniques. PMID- 12900606 TI - Patterns of initial recurrence and prognosis after sentinel lymph node biopsy and selective lymphadenectomy for melanoma. AB - The histologic status of the sentinel lymph node is a highly significant prognostic factor for patients with clinically localized cutaneous melanoma. The patterns of initial treatment failure of patients with positive sentinel lymph node biopsy versus those with negative results have not been well described. The purpose of this study was to determine the relative prognostic importance of sentinel lymph node status and to compare patterns of initial treatment failure and prognosis of node-positive versus node-negative cutaneous melanoma patients staged by sentinel lymph node biopsy and selective lymphadenectomy. The authors reviewed the pertinent demographic and surgical data in a consecutive series of patients with cutaneous melanoma who underwent sentinel lymph node staging of nonpalpable regional nodes. Sentinel lymph node biopsy was performed using a combination of blue dye and radiolocalization. Patients with positive biopsy results underwent selective lymphadenectomy, whereas those with negative results were observed. Site(s) and date(s) of initial recurrence and death were determined, and disease-free and overall survival probabilities were compared between positive and negative groups using the log-rank test and multivariable Cox regression analysis. Between February of 1994 and August of 2000, 408 patients with melanoma underwent sentinel lymph node biopsy to stage 518 regional lymph node basins. Mean Breslow tumor thickness was 2.27 mm (range, 0.2 to 14.0 mm). Eighty-five patients (20.8 percent) had at least one histologically positive sentinel lymph node, and selective lymphadenectomy yielded additional positive lymph nodes in 18 of 84 patients (21.4 percent). Recurrences were noted in 70 patients (17 percent) at a median follow-up period of 31.4 months. Recurrences were more frequent in patients with positive biopsy results (36.5 percent) than in those with negative results (12.1 percent, p < 0.0001). Distant sites of initial recurrence were more likely in the positive group than in the negative group (71 percent versus 49 percent of recurrences, respectively; p = 0.06). The false-negative rate for sentinel lymph node staging was 4.5 percent and overall accuracy was 99 percent compared with clinical follow-up. Disease-free and overall survival correlated significantly with tumor thickness, ulceration, sentinel lymph node status, and the number of tumor-positive lymph nodes (two sided p < 0.0001 for all comparisons). Multivariable analysis revealed that sentinel lymph node status (p = 0.003), tumor thickness (p = 0.016), ulceration (p = 0.006), and age (p = 0.003) were significant independent predictors of survival for the entire group. Tumor thickness and ulceration were significant predictors of recurrence and survival in sentinel node-negative patients but not in sentinel node-positive patients. Sentinel lymph node histology is possibly the most important negative predictor of early recurrence and survival in patients with American Joint Committee on Cancer stage I and II melanoma. The number of positive lymph nodes provides additional prognostic information. Although sentinel node-negative patients are a prognostically favorable group, various combinations of local and regional recurrences comprise the most common pattern of initial relapse after a negative sentinel lymph node biopsy result. PMID- 12900607 TI - Early experience with computed tomographic angiography in microsurgical reconstruction. AB - Preoperative angiography is frequently used in the planning of microsurgical reconstruction. However, several potentially devastating complications can result from angiography, including arterial occlusion and pseudoaneurysm. Computed tomographic angiography is a relatively new technique that can provide detailed information about vascular anatomy as well as soft and bony tissue without the risks of traditional angiography. In addition, three-dimensional image reconstruction uniquely demonstrates anatomical relationships among blood vessels, bones, and soft tissue. Fourteen computed tomographic angiograms were obtained in 10 patients undergoing microsurgical reconstruction of the head and neck, lower extremity, or upper extremity. The average patient age was 46.9 years (range, 22 to 67 years). Charges related to the computed tomographic procedure were compared with those of conventional preoperative imaging for microsurgical repair. At our institution, the average computed tomographic angiogram charge was 1140 US dollars, whereas the average charge for traditional arteriography was 3900 US dollars. When compared with intraoperative evaluation, computed tomographic angiograms demonstrated clinically relevant surgical anatomy. No complications were noted for the radiographic procedure or after free flap reconstruction. Computed tomographic angiography provides high-resolution, three dimensional arterial, venous, and soft-tissue imaging without the risks of traditional angiogram and at a lower cost. PMID- 12900608 TI - The role of magnetic resonance imaging in the management of vascular malformations of the trunk and extremities. AB - Vascular malformations can usually be diagnosed on clinical grounds. They have a well-defined appearance on magnetic resonance imaging, which can effectively determine their tissue and flow characteristics. However, the role of cross sectional imaging in the management of vascular malformations is not well defined. Most reviews suggest that magnetic resonance imaging should be reserved for cases in which the extent of the lesion cannot be estimated on physical examination. However, to date no group has compared the accuracy of physical examination alone to that of magnetic resonance imaging in determining this extent. A review was performed of all the patients evaluated for vascular malformations at the New York University Trunk and Extremity Vascular Anomalies Conference between July of 1994 and August of 1999. Patients who underwent magnetic resonance evaluation at other institutions and whose images were not available for review were excluded. All study patients either underwent magnetic resonance imaging examination at New York University Medical Center or had outside films reviewed at the center. The physical examination findings were compared with the magnetic resonance findings and the surgeon and radiologist made a joint decision about whether there was a correlation between the magnetic resonance and physical examination findings. Fifty-eight patients met the study criteria, 44 (76 percent) of whom were found to have more extensive disease on magnetic resonance examination than appreciated on physical examination. Of the 51 patients with low-flow vascular malformations (venous vascular malformations, lymphatic malformations, and capillary malformations), 39 (76 percent) had more extensive disease on magnetic resonance examination than on physical examination. Of the seven patients with high-flow arteriovenous malformations, five had more extensive disease on magnetic resonance. In all of the 44 patients whose magnetic resonance imaging findings did not correlate with those of the physical examination, therapeutic decision making was affected. Contrary to the conventional wisdom of published reviews, physical examination findings significantly underestimated the extent of vascular malformations in the majority of cases. Magnetic resonance imaging should be performed in all patients with vascular malformations of the trunk and extremities before therapy is planned. In an age when physicians are asked to justify their decisions, especially where the use of expensive diagnostic modalities is concerned, the situations in which these tests are indispensable must be clearly defined or else patients will be denied access to them. PMID- 12900609 TI - Anatomic study of vaginal width in male-to-female transsexual surgery. AB - Despite the recent improvement in the design of male-to-female sex reassignment operations to enlarge the vaginal vault and depth, the size of the neovagina remains somewhat limited and the exterior of the neovagina may be compressed by the bony structure of pubic rami. The purpose of this study was to determine by anatomic study the possible cause of this limitation. Eighteen male and 10 female cadavers were dissected to measure the distance between the bilateral bony pubic rami (interramic distance) at a level that corresponds to the same level in the vaginal canal of females. At the same level of the vaginal canal in the female, which corresponds to the lower border of the prostate in the male, empirically 3 cm below the bony symphysis pubis, the mean value of the interramic distance was 3.95 +/- 0.25 cm in the male and 5.20 +/- 0.36 cm in the female (p = 0.000). The interramic distance in the male and female is significantly different. In those who have undergone the male-to-female transsexual operation, the newly structured vagina may be ventrolaterally limited. Several factors cause narrowing of the vaginal orifice in male-to-female transsexuals. On the basis of this study, it seems that the bony structure of the pubic rami compresses the vagina ventrolaterally. This finding may suggest refinements of the structural design of the neovagina and prompt procedural changes in male-to-female sex reassignment operations. Future investigation should be directed toward modifying vaginoplasty so that neovaginal width can be increased to the patient's satisfaction. PMID- 12900610 TI - Osteogenesis in calvarial defects: contribution of the dura, the pericranium, and the surrounding bone in adult versus infant animals. AB - Guided bone regeneration is a promising means for reconstructing bone defects in the cranium. The present study was performed to better define those factors that affect osteogenesis in the cranium. The authors studied a single animal model, investigating the contribution of the dura, the pericranium, and the adjacent calvarial bone in the process of calvarial regeneration in both mature and immature animals. Bilateral, 100-mm2, parietal calvariectomies were performed in immature (n = 16) and mature (n = 16) rabbits. Parietal defects were randomized to one of four groups depending on the differential blockade of the dura and/or the pericranium by expanded polytetrafluoroethylene membranes. Animals were humanely killed after 12 weeks, and histometric analysis was performed to quantitate the area of the original bone defect, new bone formation, and new bone density. Bone formation was quantified separately both at the periphery and in the center of the defects. Extrasite bone formation was also quantified both on the dural and on the pericranial sides of the barriers. Bone regeneration was incomplete in all groups over the 12-week study period, indicating that complete bone healing was not observed in any group. The dura was more osteogenic than the pericranium in mature and immature animals, as there was significantly more extrasite bone formed on the dural side in the double expanded polytetrafluoroethylene barrier groups. In both the dural and the double expanded polytetrafluoroethylene barrier groups, dural bone production was significantly greater in immature compared with mature animals. The dura appeared to be the source of central new bone, because dural blockade in the dural and double expanded polytetrafluoroethylene groups resulted in a significant decrease in central bone density in both mature and immature animals. Paradoxically, isolation of the pericranium in mature animals resulted in a significant reduction in total new bone area, whereas pericranial contact appeared to enhance peripheral new bone formation, with the control group having the greatest total new bone area. The present study establishes a model to quantitatively study the process of bone regeneration in calvarial defects and highlights differences in the contribution of the dura and pericranium to calvarial bone regeneration between infant and adult animals. On the basis of these findings, the authors propose that subsequent studies in which permeability of the expanded polytetrafluoroethylene membranes is altered to permit migration of osteoinductive proteins into the defect while blocking prolapse of adjacent soft tissues may help to make guided bone regeneration a realistic alternative for the repair of cranial defects. PMID- 12900611 TI - FGF-2 stimulation affects calvarial osteoblast biology: quantitative analysis of nine genes important for cranial suture biology by real-time reverse transcription polymerase chain reaction. AB - Appropriately timed closure of the cranial sutures is a critical factor in normal postnatal morphogenesis of the cranial vault. Suture patency is necessary to permit rapid neonatal expansion of the cerebral hemispheres, and later ossification is important for bony protection of the cerebrum. Premature suture ossification (craniosynostosis) leads to myriad adverse functional and developmental consequences. Several murine studies have implicated dura-derived fibroblast growth factor-2 (FGF-2) paracrine signaling as a critical factor promoting physiologic posterior frontal suture fusion. In this study, the authors used real-time reverse transcription polymerase chain reaction (RT-PCR) to study an in vitro system that models the in vivo stimulation of suture calvarial osteoblasts by dura-derived FGF-2. The authors advocate real-time RT-PCR as a powerful and rapid technique that offers advantages in the highly sensitive, specific, and reproducible analyses of nine genes known to be important in cranial suture biology. The genes studied were growth factors [FGF-2, transforming growth factor (TGF)-beta 1, TGF-beta 2, and TGF-beta 3], growth factor receptors (FGF-R1, FGF-R2, TGF-beta RI, and TGF-beta RII), and a marker of osteoblast differentiation (Co1-I alpha I). These analyses provide a "snapshot" of several important genes involved in suture fusion that is more inclusive and quantitative than that which has been previously reported. PMID- 12900612 TI - Comparison of the biological activities of high-density porous polyethylene implants and oxidized regenerated cellulose-wrapped diced cartilage grafts. AB - The use of alloplastic materials in plastic surgery has become more extensive with advancement of autogenous-tissue reconstruction techniques for the repair of defects, tissue augmentation, and the stabilization of bones. An ideal alloplastic material should be nonallergenic, noncarcinogenic, sterilizable, and easy to shape and should not cause rejection. Alloplastic material used for tissue augmentation should have a low rate of resorption and distortion. High density porous polyethylene implants (Medpor) have been used widely and successfully for tissue augmentation. The Turkish Delight is a material composed of diced cartilage grafts wrapped in oxidized regenerated cellulose (Surgicel). Its indications are similar to those of the Medpor implant, and an additional donor site is usually not needed. Both materials are used in the same anatomical locations, especially for augmentation. Therefore, the authors evaluated the long term stability of and suitable anatomical sites for these materials. Medpor implants or Turkish Delights were placed subperiosteally or subfascially in 10 young rabbits, and the resultant changes were evaluated 16 weeks after the operation by macroscopy and histopathology. Changes in projections were measured with an ocular micrometer. Medpor implants were neither resorbed nor distorted when placed subperiosteally or subfascially, and were highly stabilized by the surrounding tissues. Turkish Delight also enabled tissue augmentation, but had a significantly higher rate of resorption compared with the Medpor implant and was loosely bound to the surrounding tissue. The Turkish Delight was less resorbed and better fixed to adjacent tissues when placed subperiosteally than when placed subfascially. PMID- 12900613 TI - Reliable assessment of skin flap viability using orthogonal polarization imaging. AB - Intraoperative evaluation of skin flap viability has primarily been dependent on clinical judgment. The purpose of this study was to determine whether an orthogonal polarization spectral imaging device could be used to accurately predict viability of random-pattern skin flaps. Orthogonal polarization spectral imaging is a newly developed technique that visualizes the microcirculation using reflected light without the use of fluorescent dyes and allows for noninvasive real-time observation of functional microvascular networks. In Sprague-Dawley rats (n = 24), three types of random skin flaps were designed with unknown zones of viability (n = 8 per group). After flap elevation, the skin flaps were evaluated by both clinical examination and orthogonal polarization spectral imaging. Areas of the flap determined to be nonviable by clinical examination were measured and marked. Orthogonal polarization spectral imaging was subsequently performed, and areas of the skin flap with stasis (i.e., cessation of red blood cell movement) in the dermal microcirculation on orthogonal polarization spectral imaging were measured and marked. The skin flaps were then secured in place. Flaps were evaluated on a daily basis for clinical signs of ischemia and necrosis. On postoperative day 7, the total amount of random skin flap necrosis was measured and recorded. Clinical examination of the random skin flaps significantly underestimated the actual amount of eventual flap necrosis, and as result was a very poor predictor of flap necrosis. By contrast, assessment of microcirculatory stasis using the orthogonal polarization spectral imaging device correlated well with the subsequent development of necrosis in all groups. In the three groups, the average amount of flap necrosis predicted by clinical examination deviated from actual necrosis by approximately 2 to 4 cm. However, the amount that orthogonal polarization spectral imaging differed from actual necrosis was 0.1 to 0.3 cm. Therefore, orthogonal polarization spectral imaging was an excellent predictor of eventual flap necrosis and much more accurate than clinical observation (p < 0.001). Intraoperative evaluation of axial and random pattern flap viability has traditionally been based on clinical examination as no other reliable, convenient test currently exists. The authors demonstrated that an orthogonal polarization spectral imaging device accurately predicts zones of necrosis in random pattern flaps by directly visualizing cessation of microcirculatory flow. Intraoperative stasis in the dermal microcirculation correlated precisely with subsequent flap necrosis. Orthogonal polarization spectral imaging was significantly more accurate than clinical examination, which consistently underestimated flap necrosis. The orthogonal polarization spectral imaging technique may have value in the intraoperative assessment of skin flap perfusion such as that required after skin-sparing mastectomy. PMID- 12900614 TI - Evaluation of the mechanism of vascular endothelial growth factor improvement of ischemic flap survival in rats. AB - This study evaluated the effects of exogenous vascular endothelial growth factor (VEGF) on the regulation of cytokines in a rat dorsal ischemic skin flap model. Exogenous VEGF (1 microg/ml) was injected subdermally into the flaps of 12 rats before the flaps were sutured back in place. Another 12 rats with flaps received saline injections, as a control group. Biopsy specimens were obtained from the flaps treated with VEGF or saline solution, at positions 2.5, 5.5, and 8.5 cm from the distal edge of the flaps, at 12 hours (n = 6 for each group) and 24 hours (n = 6 for each group) after suturing of the flaps. Expression of cytokine, growth factor, and inducible nitric oxide synthase was measured. The results demonstrated that expression of tumor necrosis factor-alpha and nitric oxide synthase in the distal part of the VEGF-treated flaps was significantly decreased, compared with the control values, at 12 and 24 hours postoperatively. It was concluded that administration of exogenous VEGF could protect flaps from ischemia-reperfusion injury through the regulation of proinflammatory cytokines and the inhibition of cytotoxic nitric oxide production. PMID- 12900615 TI - Small intestinal submucosa in abdominal wall repair after TRAM flap harvesting in a rat model. AB - The strength of porcine small intestinal submucosa in abdominal wall repair after transverse rectus abdominis myocutaneous flap harvesting was examined in a rat model. Changes in the levels of selected molecular markers of inflammation after small intestinal submucosa implantation were also studied. Eighty-three rats were divided into three groups. In experimental group I, an abdominal wall defect created by removal of the rectus abdominis muscle was repaired with placement of a 1.5 x 5-cm2 patch of small intestinal submucosa. In experimental group II, the muscle defect was repaired with a combination of small intestinal submucosa patch placement and fascial closure. In the control group, the defect was repaired with direct fascial closure. At postoperative times of 3 days, 2 weeks, 1 month, and 2 months, the muscle tissues adjacent to the abdominal wall repair site were subjected to biopsies for assessment of inflammation markers. Full-thickness sections of the abdominal wall from the repair site in each animal were removed for tensile strength testing and histological examinations. The results demonstrated that interleukin-6 and interferon-gamma levels were increased in the two experimental, small intestinal submucosa-treated groups at 3 days and 2 weeks postoperatively. The results of mechanical testing demonstrated that the average tensile strength of the repaired abdominal wall in the repair model with combined small intestinal submucosa placement and fascial repair was significantly greater than the values for repairs with fascial closure or small intestinal submucosa placement alone. The use of small intestinal submucosa placement in combination with fascial repair can significantly improve the strength of the repaired abdominal wall after transverse rectus abdominis myocutaneous flap harvesting. PMID- 12900616 TI - Dorsal scapular breast in a woman. PMID- 12900617 TI - A novel approach to correction of the prominent lobule during otoplasty. PMID- 12900618 TI - Apligraf in the treatment of severe mitten deformity associated with recessive dystrophic epidermolysis bullosa. PMID- 12900619 TI - Simultaneous repair of two adjacent digital defects with a longitudinally split reverse radial forearm flap. PMID- 12900620 TI - The ten test revisited. PMID- 12900621 TI - Tensor fasciae latae flap: long-term follow-up and refinement of its application. PMID- 12900622 TI - Hepatitis C: a review. AB - Hepatitis C virus is an RNA virus in the Flavivirus family that was identified in 1989. Since then, blood donor screening has reduced the incidence of acute infections; however, because this virus frequently leads to asymptomatic chronic infection, the prevalence of infection remains high. Chronic infection leads to increased risks of cirrhosis and hepatocellular carcinoma, as well as extrahepatic manifestations. Guidelines for widespread screening continue to evolve, and early diagnosis is likely to become more important with the development of more effective treatments. Current recommendations regarding screening are reviewed. PMID- 12900623 TI - Common craniofacial anomalies: facial clefts and encephaloceles. AB - The wide variety of craniofacial malformations makes classification difficult. A simple classification system allows an overview of the current understanding of the causes, assessments, and treatments of the most frequently encountered craniofacial anomalies. Facial clefts and encephaloceles are reviewed with respect to their diverse causes, pathogenesis, anatomical features, and treatments. Approaches to the surgical treatment of these conditions are reviewed. PMID- 12900624 TI - The role of prophylactic antibiotics in plastic surgery: whom are we treating? PMID- 12900625 TI - Measuring outcomes in plastic surgery: body image and quality of life in abdominoplasty patients. AB - Abdominoplasty is an increasingly common aesthetic surgery procedure that has yet to be evaluated using the most recently developed and psychometrically sophisticated measures of body image and quality-of-life outcomes. This study prospectively evaluated 30 consecutive female abdominoplasty patients, preoperatively and postoperatively, using measures of body image, psychological investment in appearance, and general psychosocial functioning. One-way repeated measures (pretest versus posttest) analyses of variance revealed significant positive postsurgical changes on the Appearance Evaluation subscale of the Multidimensional Body-Self Relations Questionnaire. Mean scores for the questionnaire's Body Areas Satisfaction Scale improved postoperatively (p < 0.001). Scores for the Body Exposure and Sexual Relations Questionnaire also improved significantly (p < 0.001) postoperatively. These findings indicate significant improvements in body image outcome, including positive changes in patients' evaluations of their overall appearance, their average body image dissatisfaction, and their experiences of self-consciousness and avoidance of body exposure during sexual activities. As predicted, no changes were seen on any measure of psychological investment in appearance or on patients' reports of general psychosocial functioning (self-esteem, satisfaction with life, or social anxiety). PMID- 12900626 TI - Endoscopic brow lift: a retrospective review of 628 consecutive cases over 5 years. AB - Since its introduction in 1992, endoscopic brow lift has gained tremendous recognition because it has been promoted as a novel technique to correct brow ptosis as well as glabella rhytids in a minimally invasive manner with fewer complications than the classic coronal brow lift method. In this retrospective study, 628 endoscopic brow lift procedures performed over a 5-year period (1997 2001) at Manhattan Eye Ear and Throat Hospital were reviewed. The number of endoscopic brow lift procedures performed at this institution has declined 70 percent. The purpose of this study was to elucidate the causes of this striking trend by soliciting the opinions of 21 New York plastic surgeons on their current brow ptosis management. The response rate was 84 percent (21 of 25 surgeons contacted). Currently, 25 percent of the interviewed plastic surgeons perform endoscopic brow lift regularly, 50 percent of the plastic surgeons perform endoscopic brow lift occasionally, and 25 percent of the participants no longer perform endoscopic brow lift. While most patients (70 percent) were satisfied with their results, only 50 percent of the plastic surgeons were pleased with the long-term results (after more than 2 years of follow-up). Observed postsurgical complications of endoscopic brow lift included alopecia, hairline changes, infected hardware, brow asymmetry requiring surgical revision, prolonged forehead/brow paresthesia, frontal branch nerve paralysis, and scalp dysesthesia. These complications were similar to those resulting from open brow lifts. Seventy one percent of the surveyed New York plastic surgeons routinely administered botulinum toxin type A (Botox) within 6 months of the endoscopic brow lift procedure. Possible explanations for the decline in the overall number of endoscopic brow lift procedures include the following: (1) the selection criteria for the ideal endoscopic brow lift patients are currently more limited; (2) other techniques equal or surpass endoscopic brow lift in effectiveness and predictability; and (3) endoscopic brow lift is ineffective in the majority of patients. There is no single superior surgical procedure for brow ptosis management available at this time. PMID- 12900627 TI - Safety and efficacy in an accredited outpatient plastic surgery facility: a review of 5316 consecutive cases. AB - Advances in medicine have improved the delivery of health care, making it more technologically superior than ever and, at the same time, more complex. Nowhere is this more evident than in the surgical arena. Plastic surgeons are able to perform procedures safely in office-based facilities that were once reserved only for hospital operating rooms or ambulatory surgery centers. Performing procedures in the office is a convenience to both the surgeon and the patient. Some groups have challenged that performing plastic surgery procedures in an office-based facility compromises patient safety. Our study was done to determine whether outcomes are adversely affected by performing plastic surgery procedures in an accredited outpatient surgical center. A retrospective review was performed on 5316 consecutive cases completed between 1995 and 2000 at Dallas Day Surgical Center, Dallas, Texas, an outpatient surgical facility. Most cases were cosmetic procedures. All cases were analyzed for any potential morbidity or mortality. Complications requiring a return to the operating room were determined, as were infection rates. Events leading to inpatient hospitalization were also included. During this 6-year period, 35 complications (0.7 percent) and no deaths were reported. Most complications were secondary to hematoma formation (77 percent). The postoperative infection rate for patients requiring a return to the operating room was 0.11 percent. Seven patients required inpatient hospitalization following their procedure secondary to arrhythmias, angina, and pulmonary emboli. Patient safety must take precedence over cost and convenience. Any monetary savings or time gained is quickly lost if safety is compromised and complications are incurred. The safety profile of the outpatient facility must meet and even exceed that of the traditional hospital-based or ambulatory care facility. After reviewing our experience over the last 6 years that indicated few complications and no deaths, we continue to support the judicious use of accredited outpatient surgical facilities by board-certified plastic surgeons in the management of plastic surgery patients. PMID- 12900628 TI - A morphometric study of the external ear: age- and sex-related differences. AB - The human ear is a defining feature of the face. Its subtle structures convey signs of age and sex that are unmistakable yet not easily defined. With analysis of normative cross-sectional data, this study explored anatomic and aesthetic differences in the ear between men and women, as well as changes in ear morphology with age. A total of 123 volunteers were randomly selected for this study. The cohort consisted of 89 women ages 19 to 65 years (median age, 42 years) and 34 men ages 18 to 61 years (median age, 35 years). The average total ear height across the entire cohort for both left and right ears was 6.30 cm, average lobular height was 1.88 cm, and average lobular width was 1.96 cm. As expected based on head size, significant sex-related differences were noted in the distance from the lateral palpebral commissure to both the helical root and insertion of the lobule. Measured distances in both vectors were approximately 4.6 percent longer in men than in women. Similarly, the height of the pinna was significantly larger in men than in women by approximately 6.5 percent. The average height and width of the lobule, however, were nearly identical in men and women. Analysis of age-related data showed a significant difference in the total ear height between the subpopulations; however, this difference was not significant after the lobular height was subtracted from total ear height, suggesting that the lobule was the only ear structure that changed significantly with age. In addition, lobular width decreased significantly with age. This study establishes normative data for ear morphology and clearly demonstrates the changes in earlobe morphology that occur with advancing age. PMID- 12900629 TI - Aesthetic eyelid ptosis correction: a review of technique and cases. AB - Upper eyelid ptosis can present both functional and aesthetic problems. Because proper correction of ptosis can be difficult to achieve, numerous surgical procedures have been developed. Plication of levator aponeurosis can be combined with aesthetic blepharoplasty and facial rejuvenation procedures to successfully address ptosis. The authors assessed the effectiveness of levator aponeurosis plication for correction of acquired upper eyelid ptosis in patients presenting for concomitant cosmetic facial procedures. The medical records of 74 consecutive patients (68 women and six men) who had upper eyelid ptosis correction in conjunction with cosmetic facial procedures from January of 1994 to January of 2000 were reviewed. During this period, 400 endoscopic forehead lifts and 479 face lifts were performed. The correction was performed through an external upper blepharoplasty approach removing an ellipse of skin and orbicularis muscle. Once the orbital septum was opened, a plication of the levator aponeurosis was accomplished by one or more horizontal mattress sutures of 6-0 clear nylon (with the first bite placed at or just medial to the vertical level of the pupil). The average follow-up period was 14 months. Long-term correction of the ptosis was excellent. The complications were minor, with the most common occurrence being asymmetry. Revisions were performed on only four patients. Correction of ptosis can be performed safely and effectively in conjunction with periorbital and facial rejuvenation. The technique described is simple, reliable, and reproducible. PMID- 12900630 TI - Short-scar face lift with extended SMAS platysma dissection and lifting and limited skin undermining. AB - In the past 8 years, short-scar cervicofacial rhytidectomy has been used with great success and a high level of patient satisfaction. The operation involves limited incisions in the sideburn and preauricular areas, extending just around the fold of the ear lobule. Extensive undermining of the superficial musculoaponeurotic system (SMAS)/platysma is performed, and lifting of the face and neck is performed mainly at this level. Skin undermining is kept to a minimum, eliminating the need for extended incisions to reduce dog-ears. The preservation of fibrous bands between the skin and the SMAS/platysma unit is an important factor in making such an operation practical and is the reason why the skin incisions can be kept short. Dissection of large areas of skin and separation of the skin from the underlying SMAS necessitate larger incisions. The operation is safe, speedy, less traumatic, and well tolerated among patients at high risk, such as smokers. The risks of complications are also reduced with this operation. PMID- 12900631 TI - Correction of thin lips: a 17-year follow-up of the original technique. PMID- 12900633 TI - The work. PMID- 12900632 TI - Labial incompetence: a marker for progressive bone resorption in silastic chin augmentation: an update. PMID- 12900634 TI - Of chickens and red flags. PMID- 12900636 TI - An anatomic study and clinical applications of the reversed submental perforator based island flap. PMID- 12900637 TI - Limited incision for breast surgery. PMID- 12900639 TI - A novel skin incision for digital access. PMID- 12900638 TI - Transposing a chest wall flap to the breast. PMID- 12900640 TI - Arnica montana and homeopathic dosing guidelines. PMID- 12900641 TI - The use of epinephrine in breast reduction. PMID- 12900642 TI - Improved subcutaneous mastectomy with hydrodissection of the subcutaneous space. PMID- 12900643 TI - Scalpel caliper. PMID- 12900644 TI - Tram flap breast reconstruction and weight fluctuations: it is alive! PMID- 12900645 TI - Sickle cell trait: a risk factor for flap necrosis. PMID- 12900646 TI - Treatment of breast asymmetry with free adiposal flap from the lower abdomen: unsuccessful attempt. PMID- 12900647 TI - The deviated nose: optimizing results using a simplified classification and algorithmic approach. PMID- 12900648 TI - A new ear reduction method without an anterior scar: posterior approach. PMID- 12900649 TI - The advantage of the anterolateral thigh flap for reconstruction of the anterior skull base defect in recurrent cases. PMID- 12900650 TI - Technique for optimizing exposure in great toe revascularization and replantation. PMID- 12900651 TI - Gigantic metameric seborrheic keratosis. PMID- 12900652 TI - An interesting etiologic reason for skin defect in plastic surgery: air bag deployment. PMID- 12900653 TI - The reproducible tie-over dressing. PMID- 12900654 TI - Is the hypoglossal nerve a reliable donor nerve for transfer in brachial plexus injuries? PMID- 12900655 TI - Ankle-arm index versus angiography for preassessment of the fibula free flap. PMID- 12900656 TI - Intertask transfer in plastic surgery. PMID- 12900657 TI - Arm restraint in children with cleft lip and palate. PMID- 12900658 TI - Incisional endometrioma. PMID- 12900659 TI - A long-lasting pyogenic granuloma. PMID- 12900661 TI - Anatomic facial retractors. PMID- 12900660 TI - No need for drainage after reduction mammaplasty. PMID- 12900662 TI - A "splash" twist to a painless breast augmentation. PMID- 12900663 TI - Removal of perforated silicone implants: how to avoid a sticky operational field. PMID- 12900668 TI - What's different about heart disease in women? PMID- 12900669 TI - Keeping West Nile virus at bay. PMID- 12900670 TI - Without warning. PMID- 12900671 TI - Nailing a key assessment. Learn the significance of certain nail anomalies. AB - Discover how a patient's fingernails can point out specific health problems. PMID- 12900672 TI - When your patient needs peritoneal dialysis. Brush up on this necessary but infrequently used skill that you may need if your patient has chronic renal failure. PMID- 12900673 TI - New drugs 2003, part III. PMID- 12900674 TI - Epidural hematoma. PMID- 12900675 TI - ACSM and the emergence of the profession of exercise physiologist. PMID- 12900676 TI - The human gene map for performance and health-related fitness phenotypes: the 2002 update. AB - This review presents the 2002 update of the human gene map for physical performance and health-related phenotypes. It is based on peer-reviewed papers published by the end of 2002 and includes association studies with candidate genes, genome-wide scans with polymorphic markers, and single gene defects causing exercise intolerance to variable degrees. The genes and markers with evidence of association or linkage with a performance or fitness phenotype in sedentary or active people, in adaptation to acute exercise, or for training induced changes are positioned on the genetic map of all autosomes and the X chromosome. Negative studies are reviewed, but a gene or locus must be supported by at least one positive study before being inserted on the map. By the end of 2000, 29 loci were depicted on the map. The 2001 map includes 71 loci on the autosomes and two on the X chromosome. In contrast, the 2002 human gene map for physical performance and health-related phenotypes includes 90 gene entries and QTL, plus two on the X chromosome. To all these loci, one must add 14 mitochondrial genes in which sequence variants have been shown to influence relevant fitness and performance phenotypes. PMID- 12900677 TI - Resistance training on physical performance in disabled older female cardiac patients. AB - PURPOSE: We evaluated the value of resistance training on measures of physical performance in disabled older women with coronary heart disease (CHD). METHODS: The study intervention consisted of a 6-month program of resistance training in a randomized controlled trial format. Training intensity was at 80% of the single repetition maximal lift. Control patients performed light yoga and breathing exercises. Study participants included 42 women with CHD, all >or= 65 yr of age and community dwelling. Subjects were screened by questionnaire to have low self reported physical function. The primary study measurements related to the performance of 16 household activities of the Continuous Scale Physical Functional Performance test (CSPFP). These ranged from dressing, to kitchen and cleaning activities, to carrying groceries and walking onto a bus with luggage, and a 6-min walk. Activities were measured in time to complete a task, weight carried during a task, or distance walked. Other measures included body composition, measures of aerobic fitness and strength, and questionnaire-based measures of physical function and depression score. RESULTS: Study groups were similar at baseline by age, aerobic capacity, strength, body composition, and in performing the CSPFP. After conditioning, 13 of 16 measured activities were performed more rapidly, or with increased weight carried, compared with the control group (all P < 0.05). Maximal power for activities that involved weight bearing over a distance, increased by 40% (P < 0.05). CONCLUSIONS: Disabled older women with CHD who participate in an intense resistance-training program improve physical capacity over a wide range of household physical activities. Benefits extend beyond strength-related activities, as endurance, balance, coordination, and flexibility all improved. Strength training should be considered an important component in the rehabilitation of older women with CHD. PMID- 12900679 TI - Sedentary lifestyle, poor cardiorespiratory fitness, and the metabolic syndrome. AB - PURPOSE: The cross-sectional associations of leisure-time physical activity (LTPA) and cardiorespiratory fitness with the metabolic syndrome (MS) were investigated in a population-based sample of 1069 middle-aged men without diabetes, cardiovascular disease, or cancer. METHODS: LTPA was assessed using a detailed quantitative questionnaire. Maximal oxygen uptake VO(2max) and core and related features of the MS were determined. A modified World Health Organization definition of the MS and factor analysis were used. RESULTS: Men who engaged in at least moderate-intensity (>or=4.5 metabolic equivalents) LTPA <1.0 h.wk-1 were 60% more likely to have the MS than those engaging in >or= 3.0 h.wk-1 even after adjustment for confounders. Low-intensity (<4.5 metabolic equivalents) LTPA was not associated with the metabolic syndrome. Men with a VO(2max) <29.1 mL x kg-1 x min-1 were almost seven times more likely to have the MS than those with a VO(2max) >or=35.5 mL.kg-1.min-1 even after adjusting for confounders. In first order factor analysis using a promax rotation, the principal factor explained 20% of total variance and had heavy loadings for VO(2max) (-0.57) and at least moderate-intensity LTPA (-0.44), and moderate or heavy loadings for the main components of the MS. The second-order factor analysis generated a principal factor that was strongly loaded on by the main components of the MS and VO(2max) (-0.50) but not LTPA. CONCLUSION: A sedentary lifestyle and especially poor cardiorespiratory fitness are not only associated with the MS but could also be considered features of the MS. Measurement of VO(2max) in sedentary men with risk factors may provide an efficient means for targeting individuals who would benefit from interventions to prevent the MS and its consequences. PMID- 12900680 TI - Genetic correlation of exercise with heart rate and respiratory sinus arrhythmia. AB - PURPOSE: A twin design was used to test whether the association between exercise behavior and heart rate and the association between exercise behavior and respiratory sinus arrhythmia (RSA) derive from a common genetic factor. METHODS: Data were available from 157 adolescent (aged 13-22) and 208 middle-aged twin pairs (aged 35-62), divided into five sex by zygosity groups (male and female monozygotic twin pairs, and dizygotic twin pairs of same or opposite sex). Exercise behavior was assessed as the average weekly METs spent on sports activities or other vigorous activities in leisure time (sportMETS) in the last 3 months. RSA and heart period (HP) were assessed in the time domain from the combined ECG and respiration signals. RESULTS: Heritability estimates were 16% and 29% for RSA, 64% and 68% for HP, and 79% and 41% for sportMETS in young and middle-aged twins, respectively. A significant association was found between RSA and sportMETS (0.17) in the adolescent twins that derived entirely from a common genetic factor. No association was found between sportMETS and RSA in the older twins. A significant association was found between HP and sportMETS in both adolescent (0.35) and middle-aged (0.18) twins. A large contribution of common genetic factors to these associations was found amounting to 84% and 88% in the young and middle-aged twins, respectively. CONCLUSIONS: Although the results of this study do not preclude causal effects of exercise on RSA and heart rate, they show that the association between exercise and these cardiovascular risk factors largely derives from a common genetic factor. PMID- 12900678 TI - A community-based group exercise program for persons with chronic stroke. AB - PURPOSE: The purpose of this study was to evaluate the physical and psychosocial effects of an 8-wk community-based functional exercise program in a group of individuals with chronic stroke. METHODS: Twenty-five subjects (mean age 63 yr) participated in a repeated measures design that evaluated the subjects with two baseline assessments 1 month apart, one postintervention assessment, and one retention assessment 1 month postintervention. Physical outcome measures assessed were the Berg Balance Test, 12-Minute Walk Test distance, gait speed, and stair climbing speed. Psychosocial measures assessed were the Reintegration to Normal Living Index (RNL) and Canadian Occupational Performance Measure (COPM). The 8-wk training consisted of a 60-min, 3 x wk-1 group program that focused on balance, mobility, functional strength, and functional capacity. The program was designed to be accessible by reducing the need for costly one-on-one supervision, specialized settings, and expensive equipment. RESULTS: Improvements from the exercise program were found for all physical measures and these effects were retained 1-month postintervention. Subjects with lower function improved the most relative to their initial physical status. Significant effects were found for the COPM, but not the RNL Index; however, subjects with lower RNL improved the most relative to their initial RNL Score. CONCLUSION: A short-term community-based exercise program can improve and retain mobility, functional capacity, and balance and result in a demonstrable impact upon the performance of activities and abilities that were considered meaningful to the subjects. Implementation of such community-based programs has potential for improving activity tolerance and reducing the risk for secondary complications common to stroke (e.g., falls resulting in fractures and cardiac events). PMID- 12900681 TI - Acupuncture and responses of immunologic and endocrine markers during competition. AB - INTRODUCTION: Acupuncture is used to modulate the physical well-being of athletes in Asian countries. However, there is little information on the immediate effects of acupuncture treatment on physiological or psychological responses to exercise. PURPOSE: The purpose of this study was to examine the effect of acupuncture treatment on the physical well-being of elite female soccer players during a competition period. METHODS: Subjects were divided into two groups: those who received acupuncture treatment (18.1 +/- 2.3 yr [+/-SD], N = 9) and a control group (17.7 +/- 2.8 yr, N = 12). In the treatment group, acupuncture stimulus was applied at LI 4 (Goukoku), ST 36 (Ashi-sanri) for 20 min, and ST 6 (Kyosya), LU 6 (Ko-sai) points for 15 min 4 h after the game every night during the competition period. The measured parameters included salivary secretory immunoglobulin A (SIgA) level, cortisol level in saliva, subjective rating of physical well-being, and profile of mood states (POMS). RESULTS: The following were the main results: 1). Exercise-induced decrease of salivary SIgA and increase of salivary cortisol were inhibited by acupuncture. 2). Acupuncture improved subjective rating of muscle tension and fatigue. 3). The POMS score was modulated by acupuncture. CONCLUSION: These results support the effectiveness of acupuncture for physical and mental well-being of athletes. PMID- 12900682 TI - Effects of sodium bicarbonate ingestion on prolonged intermittent exercise. AB - PURPOSE: The aim of this study was to determine the effects of sodium bicarbonate ingestion on prolonged intermittent exercise and performance. METHODS: Eight healthy male subjects (mean +/- SD: age 25.4 +/- 6.4 yr, mass 70.9 +/- 5.1 kg, height 179 +/- 7 cm, VO(2max) 4.21 +/- 0.51 L.min-1) volunteered for the study, which had received ethical approval. Subjects undertook two 30-min intermittent cycling trials (repeated 3-min blocks; 90 s at 40% VO(2max), 60 s at 60% VO(2max), 14-s maximal sprint, 16-s rest) after ingestion of either sodium bicarbonate (NaHCO(3); 0.3 g.kg-1) or sodium chloride (NaCl; 0.045 g x kg(-1). Expired air, blood lactate (BLa), bicarbonate (HCO(3)-), and pH were measured at rest, 30 and 60 min postingestion, and during the 40% VO(2max) component of exercise (4, 10, 16, and 29 min). RESULTS: After ingestion, pH increased from rest to 7.46 +/- 0.03 and 7.40 +/- 0.01 for NaHCO(3) and NaCl, respectively (main effect for time and trial; P < 0.05). Values decreased at 15 min of exercise to 7.30 +/- 0.07 and 7.21 +/- 0.06, respectively, remaining at similar levels until the end of exercise. BLa peaked at 15 min (12.03 +/- 4.31 and 10.00 +/- 2.58 mmol.L-1, for NaHCO(3) and NaCl, respectively; P > 0.05) remaining elevated until the end of exercise (P < 0.05). Peak power expressed relative to sprint 1 demonstrated a significant main effect between trials (P < 0.05). Sprint 2 increased by 11.5 +/- 5% and 1.8 +/- 9.5% for NaHCO(3) and NaCl, respectively. During NaHCO(3), sprint 8 remained similar to sprint 1 (0.2 +/- 17%), whereas a decrease was observed during NaCl (-10.0 +/- 16.0%). CONCLUSION: The results of this study suggest that ingestion of NaHCO(3) improves sprint performance during prolonged intermittent cycling. PMID- 12900683 TI - L-arginine ingestion after rest and exercise: effects on glucose disposal. AB - PURPOSE: There is considerable interest, both in health and disease, in enhancing postexercise glucose uptake and glycogen resynthesis in skeletal muscle. The amino acid, arginine, is known to stimulate insulin release and enhance glucose stimulated insulin release. METHODS: The present investigation examined whether an oral dose of L-arginine (10 g), when given with 70 g carbohydrate (CHO, in the form of simple sugars) improved factors associated with glucose disposal in previously exercised and nonexercised healthy males. The effects of different modes of activity (resistance or cycling exercise) upon these factors were also examined. RESULTS: Whole-blood glucose and serum insulin concentrations after L arginine + CHO ingestion were not significantly different from the placebo condition (glycine + CHO ingestion) in all experimental treatments (nonexercised, resistance exercise, and cycling exercise). Similarly, CHO oxidation, forearm blood flow, blood pressure, and heart rate during the postingestion period were unaffected by L-arginine + CHO consumption in all three experimental treatments. CONCLUSION: A 10-g oral dose of L-arginine was found to have no effect on blood glucose disposal in human subjects after oral CHO ingestion, either when rested or after different modes of exercise known to differentially affect glucose disposal. These results suggest that the addition of L-arginine to a CHO beverage would not augment postexercise CHO replenishment in healthy human subjects. PMID- 12900684 TI - Effects of pseudoephedrine on maximal cycling power and submaximal cycling efficiency. AB - PURPOSE: To study the effects of a therapeutic dose of pseudoephedrine on anaerobic cycling power and aerobic cycling efficiency. METHODS: Eleven healthy moderately trained males (VO (2peak) 4.4 +/- 0.8 L x min(-1) participated in a double-blinded crossover design. Subjects underwent baseline (B) tests for anaerobic (Wingate test) and aerobic (VO (2peak) test) cycling power. Subjects ingested either 60 mg of pseudoephedrine hydrochloride (D) or a placebo (P) and, after 90 min of rest, a Wingate and a cycling efficiency test were performed. During the cycling efficiency test, heart rate (HR) and VO(2) were averaged for the last 5 min of a 10-min cycle at 40% and 60% of the peak power achieved during the VO (2peak) test. RESULTS: There were no significant differences in peak power (B = 860 +/- 154, D = 926 +/- 124, P = 908 +/- 118 W), total work (B = 20 +/- 3, D = 21 +/- 3, P = 21 +/- 3 kJ), or fatigue index (B = 39 +/- 8, D = 45 +/- 5, P = 43 +/- 5%). There were no significant differences in HR at 40% power (D = 138 +/- 10, P = 137 +/- 10 beats.min-1) or 60% power (D = 161 +/- 11, P = 160 +/- 11 beats x min(-1). There were no significant differences in cycling efficiency at 40% power (D = 18.8 +/- 1.8, P = 18.5 +/- 1.8%) or 60% power (D = 20.3 +/- 2.0, P = 20.1 +/- 2.1%). CONCLUSION: A therapeutic dose of pseudoephedrine hydrochloride does not affect anaerobic cycling performance or aerobic cycling efficiency. PMID- 12900685 TI - Adiponectin responses to continuous and progressively intense intermittent exercise. AB - PURPOSE: Adiponectin is a recently discovered adipocyte protein that is lower in patients with coronary artery disease and in Type II diabetics who have insulin resistance. Regular exercise is known to be a preventative factor in the development of atherosclerosis and Type II diabetes. Acute exercise increases insulin sensitivity; however, it also increases beta-adrenergic and glucocorticoid activities that may suppress adiponectin expression. Two experiments were conducted to determine whether acute exercise affects adiponectin concentrations. METHODS: In the first experiment, six healthy male subjects completed 30 min of heavy continuous running exercise at 79% of VO (2max). In the second experiment, well-trained runners completed strenuous intermittent exercise consisting of treadmill running at 60, 75, 90, and 100% VO (2max). A resting control trial for the second experiment was also conducted. RESULTS: Glucose and insulin were not altered significantly in the first experiment, but both increased significantly (P < 0.05) in the second experiment. A significant increase (P < 0.05) in adiponectin in the first experiment was no longer significant after correction for plasma volumes shifts. In the second experiment, there were significant (P < 0.05) changes in adiponectin concentrations over time but not a significant difference between adiponectin responses in exercise and control trials. CONCLUSIONS: The data suggest that 30 min of heavy continuous running or more strenuous intermittent running does not stimulate an increase in production and release of adiponectin, and small increases in adiponectin concentrations resulting from the exercise may be attributed to normal plasma volume shifts. PMID- 12900686 TI - Effect of prolonged exercise and carbohydrate on total neutrophil elastase content. AB - PURPOSE: The purpose of the present study was twofold: first, to assess the effect of prolonged intense exercise on total neutrophil elastase content in endurance-trained cyclists and to determine whether this is associated with postexercise falls in lipopolysaccharide (LPS)-stimulated neutrophil elastase release; and second, to determine the effect of carbohydrate (CHO) ingestion during exercise on these responses. METHODS: In a randomized design, nine trained male cyclists cycled for 2 h at 75% VO(2max) on two occasions with either CHO (6.4%, i.e., 64 g x L-1) or placebo (PLA) beverage ingestion before (5 mL x kg 1), during (2 mL x kg-1), and after (5 mL x kg-1) the exercise. Venous blood samples were obtained at rest, immediately postexercise, and at 1 h postexercise. RESULTS: After exercise, CHO ingestion was associated with a higher plasma glucose concentration (P < 0.05) and fewer numbers of circulating neutrophils compared with the PLA trial (P < 0.01). Neither exercise nor CHO ingestion affected total neutrophil elastase content, yet LPS-stimulated neutrophil elastase release fell postexercise by approximately 47% on the PLA trial (P < 0.01). Values did not change significantly from preexercise on the CHO trial. CONCLUSIONS: These findings suggest that neither exercise nor CHO-beverage ingestion influences the total elastase content of neutrophils. Therefore, changes in neutrophil elastase content cannot account for the fall in LPS stimulated neutrophil elastase release after prolonged intense exercise or for the blunting of this response with CHO ingestion. PMID- 12900687 TI - The effect of age and gender on heart rate variability after endurance training. AB - PURPOSE: This research investigated the age and gender differences in cardiovascular adaptation to a standardized/quantified endurance-training program that included two taper periods. METHODS: The latter was analyzed from spectral analysis of electrocardiogram records of heart rate variability (HRV) at rest in groups of young (19-21 yr) and middle aged (40-45 yr), mixed gender groups (6 males and 6 females), pre- and poststandardized training. All subjects were recreational runners who completed the same 12-wk running program. Before, and subsequent to training, HRV was measured during supine rest and submaximal cycling. RESULTS: There was a significant decrease in heart rate both at rest (2.7 +/- 0.45 beats x min-1) and during submaximal exercise (8.1 +/- 0.67 beats x min-1) in both age groups after training. After training, total spectral power increased (560.7 +/- 308.9 ms2), as well as high-frequency power (362.3 +/- 405.5 ms2), in both age groups at rest. The young group showed a greater increase in total power (849.0 +/- 308.7 ms2) after the training program. CONCLUSION: It is concluded that a well-designed 12-wk endurance-training program will decrease resting and submaximal heart rate in both younger and older adults. The significant increase in HRV, total power, and high-frequency power in all groups after endurance training indicates that HRV measurement appears to provide an effective, noninvasive assessment of cardiovascular adaptation to aerobic training. PMID- 12900689 TI - Effect of repeated caffeine ingestion on repeated exhaustive exercise endurance. AB - PURPOSE: The purpose of this study was to examine the effect of repeated doses of caffeine on repeated exercise endurance. METHODS Nine male caffeine users performed exercise rides (ER) to exhaustion at 80% VO(2max) after ingesting a placebo, 5 mg x kg-1 of caffeine, or 2.5 mg x kg-1 of caffeine 1 h before the ER. Two ER were performed weekly on the same day once in the morning (AM) and 5 h later in the afternoon (PM). There were four treatments containing either caffeine or placebo, i.e., trial A representing 5-mg x kg-1 caffeine in the AM and 2.5-mg x kg-1 caffeine in the PM; trial B, which was placebo in both AM and PM; trial C representing 5-mg x kg-1 caffeine in the AM and placebo in the PM; and trial D representing a placebo in the AM and 5-mg x kg-1 caffeine in the PM. The order of the treatment trials was double blind and randomized. RESULTS: Caffeine ingestion significantly increased exercise time to exhaustion in the AM (trial A 24.9 +/- 10.2 min and trial C 21.8 +/- 4.9 vs trial B 18.0 +/- 6.4 min and D 17.7 +/- 4.3 min). This effect was maintained in the PM and greater than placebo (B 18.3 +/- 4.8 min) regardless of whether redosing (trial A 21.5 +/- 8.6 min) or placebo (trial C 21.0 +/- 6.8) followed the initial morning dose. Caffeine dosing in the PM (trial D 22.4 +/- 7.2 min) also increased ER after placebo trial D in the AM. CONCLUSIONS: It was concluded that redosing with caffeine after exhaustive exercise in the AM was not necessary to maintain the ergogenic effect of the drug during subsequent exercise 6 h later. PMID- 12900688 TI - CPX/D underestimates VO(2) in athletes compared with an automated Douglas bag system. AB - PURPOSE: Based on persistent reports of low oxygen consumption VO(2) from Medical Graphics CPX/D metabolic carts, we compared the CPX/D against an automated Douglas bag system. METHODS: Twelve male athletes completed three, randomized 25 min bouts (5 min at 100, 150, 200, 250, and 300 W) on a cycle ergometer with intervening 30-min rests. One bout was measured on each of the CPX/D, the CPX/D with altered software (CPX/DDelta), and an automated Douglas bag system at Flinders University (FU). The CPX/DDelta software alteration was an apparent lag time correction factor of 60 ms. RESULTS: For the CPX/D, both VO(2) and VCO(2) were significantly lower than the FU system at 100-300 W, and the relative differences ranged -10.7 to -12.0% and -7.7 to -8.2%, respectively. Altering the software approximately halved the VO(2) discrepancy between the CPX/DDelta and FU systems. When data from all five workloads were pooled, V(E) of the CPX/D (67.2 +/- 26.4 L x min-1) and CPX/DDelta (67.5 +/- 26.9 L x min-1) were significantly lower than for the FU system (70.5 +/- 27.1 L x min-1); and at 300 W, the relative differences were -4.0% and -3.4% for the CPX/D and CPX/DDelta, respectively. Altering the software changed the pooled %O(2) from 16.24 +/- 0.40% for the CPX/D to 16.04 +/- 0.39% for the CPX/DDelta, and these were significantly different than pooled data for the FU system (16.15 +/- 0.39%). CONCLUSIONS: During submaximal exercise, the CPX/D yields VO(2) values that are approximately 11% lower than the criterion system, and the source of the discrepancy does not appear to be primarily related to volume measurement. A disturbing observation is that factory defaults for the lag time use different correction factors, which vary by 60 ms and this significantly alters VO(2) and VCO(2). PMID- 12900690 TI - Genome-wide linkage scan for physical activity levels in the Quebec Family study. AB - PURPOSE AND METHODS: It is commonly recognized that there is familial aggregation for physical activity level. However, the genes and sequence variants responsible for the familial clustering have not been investigated. We performed a genome wide linkage scan based on 432 markers typed in 767 subjects from 207 families of the Quebec Family study with the aim of identifying loci affecting physical activity levels. Four physical activity level phenotypes were used. RESULTS: Promising evidence of linkage (P < 0.0023) was found for physical inactivity on chromosome 2p22-p16. Suggestive linkages (0.0023

P > 0.0001) was observed for maximal activities of enzymes of the energy production pathways. With regard to the training response, significant familial aggregation (0.05 > P < 0.0001) was observed for maximal activities of enzymes of the energy production pathways. CONCLUSION: These data provide evidence of familial aggregation for enzyme activities of the main energy metabolism pathways of the skeletal muscle in the sedentary state and in response to regular exercise. PMID- 12900692 TI - Activity levels and body mass index of children in the United States, Sweden, and Australia. AB - PURPOSE: Assess the physical activity and body mass index (BMI) levels of children in the United States, Sweden, and Australia. METHODS: A total of 1954 children, 6-12 yr old (711 American, 563 Australian, and 680 Swedish) wore sealed pedometers for four consecutive days. Height and weight measures were obtained. RESULTS: Descriptive data for step counts and BMI by sex, age, and country were calculated to determine activity levels and BMI. Three-way multivariate ANOVA for step counts and BMI between countries at each age and sex found that, in general, the Swedish children were significantly more active than the Australian and American children, and the American children were significantly heavier than the Australian and Swedish children. For boys, the mean step counts ranged from 15673 to 18346 for Sweden, 13864 to 15023 for Australia, and 12554 to 13872 for America. For girls, the mean step counts ranged from 12041 to 14825 for Sweden, 11221 to 12322 for Australia, and 10661 to 11383 for America. The activity curve is somewhat level during the preadolescent years. The rate of increase in BMI with age is much greater in the American children than in the Swedish or Australian children. The percent of American, Swedish, and Australian boys classified as overweight/obese was 33.5, 16.6, and 15.8, respectively. The percent of American, Swedish, and Australian girls classified as overweight/obese was 35.6, 16.8, and 14.4, respectively. Correlation analysis found few significant negative relationships between step counts and BMI. CONCLUSIONS: American children tend to be the least active and heaviest with the greatest rate of increase in BMI. The Swedish children are the most active group followed by Australia. Swedish and Australian children maintain lower BMI throughout their prepubescent years than do the American children who have a greater percentage who are classified as overweight. PMID- 12900693 TI - The validity of the Stanford Seven-Day Physical Activity Recall in young adults. AB - PURPOSE: To evaluate the criterion validity of the 7-Day Physical Activity Recall (7D-PAR) and factors associated with reporting error, in a sample of moderately overweight, young adult men and women. METHODS: Average total daily energy expenditure (TDEE) and physical activity energy expenditure (PAEE) from the 7D PAR were compared with the same parameters assessed by doubly labeled water in 17 men, age = 23.9 +/- 3.8 yr, and 29 women, age = 23.3 +/- 4.6 yr, who volunteered to participate in a 16-month supervised aerobic exercise trial. PAEE was estimated from the 7D-PAR and from DLW [0.9 * TDEE -resting metabolic rate (RMR) (indirect calorimetry)]. In addition, peak oxygen uptake and percent body fat were obtained. RESULTS: No significant differences in TDEE (kJ.d-1) were noted between the 7D-PAR (11825 +/- 1779) and DLW (11922 +/- 2516) for the complete sample (N = 46) or for men (7D-PAR = 13198 +/- 1638, DLW = 13885 +/- 2754) or women (7D-PAR = 11018 +/- 1323, DLW = 10771 +/- 1457. The mean PAEE from the 7D PAR was not different from DLW in the total sample (7D-PAR = 3286 +/- 502, DLW = 3508 +/- 1863) as well as in men (7D-PAR = 3650 +/- 490, DLW = 3989 +/- 2461) and women (3073 +/- 377, DLW = 3223 +/- 1360). In a regression model, PAEE, peak oxygen uptake, gender and percent fat accounted for 86% of the reporting error in total daily energy expenditure when using the 7D-PAR. CONCLUSION: The 7D-PAR provided a reasonable estimate of both the mean TDEE and PAEE in this sample; however, estimates of energy expenditure on an individual basis using the PAR were subject to considerable error. PMID- 12900694 TI - International physical activity questionnaire: 12-country reliability and validity. AB - BACKGROUND: Physical inactivity is a global concern, but diverse physical activity measures in use prevent international comparisons. The International Physical Activity Questionnaire (IPAQ) was developed as an instrument for cross national monitoring of physical activity and inactivity. METHODS: Between 1997 and 1998, an International Consensus Group developed four long and four short forms of the IPAQ instruments (administered by telephone interview or self administration, with two alternate reference periods, either the "last 7 d" or a "usual week" of recalled physical activity). During 2000, 14 centers from 12 countries collected reliability and/or validity data on at least two of the eight IPAQ instruments. Test-retest repeatability was assessed within the same week. Concurrent (inter-method) validity was assessed at the same administration, and criterion IPAQ validity was assessed against the CSA (now MTI) accelerometer. Spearman's correlation coefficients are reported, based on the total reported physical activity. RESULTS: Overall, the IPAQ questionnaires produced repeatable data (Spearman's rho clustered around 0.8), with comparable data from short and long forms. Criterion validity had a median rho of about 0.30, which was comparable to most other self-report validation studies. The "usual week" and "last 7 d" reference periods performed similarly, and the reliability of telephone administration was similar to the self-administered mode. CONCLUSIONS: The IPAQ instruments have acceptable measurement properties, at least as good as other established self-reports. Considering the diverse samples in this study, IPAQ has reasonable measurement properties for monitoring population levels of physical activity among 18- to 65-yr-old adults in diverse settings. The short IPAQ form "last 7 d recall" is recommended for national monitoring and the long form for research requiring more detailed assessment. PMID- 12900695 TI - International physical activity questionnaire: 12-country reliability and validity. PMID- 12900696 TI - Influence of ball velocity, attention, and age on response time for a simulated catch. AB - PURPOSE: The ability of a baseball infielder to respond to a batted ball may provide the best defense for avoiding injury. This study investigated the response times of young athletes performing a simulated baseball-fielding task to estimate the maximum velocity with which a baseball can leave the bat and allow a player, standing 13.7 m away, to safely respond to the approaching ball. METHODS: Fifty boys and 50 girls between the ages of 8 and 16 yr participated. Baseballs were projected at the subjects who were standing in a standardized position behind a safety net. Two components of response time, reaction time and movement time, were determined using a motion capture system. The influences of baseball velocity (26.8 and 33.5 m.s(-1) and level of attention (full attention and attention splitting) on response time for a simulated baseball-fielding task were characterized. Based on the response times for each age group, the maximum exit velocity from a baseball-bat interaction that would allow a young baseball player sufficient time to safely respond to an approaching baseball was calculated. RESULTS: The results showed that subjects had sufficient time to respond to exit velocities from 26.8 m.s-1 (8- to 9-yr-old group) to 33.5 m.s-1 (16-yr-old group). However, the accuracy of the response was negatively affected by baseball velocity. CONCLUSIONS: If the exit-velocities seen during actual competition exceed the calculated maximum exit-velocities for these age groups, then our preliminary data suggest that modifications to the game of baseball that would reduce the actual exit-velocities and serve as an effective means to reduce the potential for serious or catastrophic injury are warranted. PMID- 12900697 TI - Linear and angular head accelerations during heading of a soccer ball. AB - PURPOSE: Cognitive deficits observed in professional soccer players may be related to heading of a soccer ball. To assess the severity of a single instance of heading a soccer ball, this study experimentally and theoretically evaluated the linear and angular accelerations experienced by the human head during a frontal heading maneuver. METHODS: Accelerations were measured using a set of three triaxial accelerometers mounted to the head of each of four adult male subjects. These measurements (nine signals) were used to estimate the linear acceleration of the mass center and the angular acceleration of the head. Results were obtained for ball speeds of 9 and 12 m.s(-1) (approximately 20 and 26 mph). A simple mathematical model was derived for comparison. RESULTS: At 9 m.s(-1), peak linear acceleration of the head was 158 +/- 19 m.s(-2) (mean +/- standard deviation) and peak angular acceleration was 1302 +/- 324 rad.s(-2); at 12 m.s( 1), the values were 199 +/- 27 m.s-2 and 1457 +/- 297 rad.s-2, respectively. The initial acceleration pulses lasted approximately 25 ms. Measured head accelerations confirmed laboratory headform measurements reported in the literature and fell within the ranges predicted by the theoretical model. CONCLUSIONS: Linear and angular acceleration levels for a single heading maneuver were well below those thought to be associated with traumatic brain injury, as were computed values of the Gadd Severity Index and the Head Injury Criterion. However, the effect of repeated acceleration at this relatively low level is unknown. PMID- 12900698 TI - The curvature constant parameter of the power-duration curve for varied-power exercise. AB - INTRODUCTION: The tolerable duration (t) for high-intensity cycle ergometry bears a hyperbolic relationship to the power output (P) with an asymptote termed the critical power (CP), and a curvature constant (W') that is numerically equivalent to an amount of work that can be performed above CP. The physiological nature of W' has received little consideration compared with CP, e.g., whether the total amount of work above CP remains constant when the power actually changes during the high-intensity task. PURPOSE: The purpose of this study was to compare W' derived from the standard estimation method, consisting of several different constant-P tests, and the total amount of work above CP during an exhausting exercise bout using a variable-P protocol. METHODS: Eleven healthy male subjects (age: 21-40 yr) volunteered to participate in this study. Each initially performed four-to-six high-intensity square-wave exercise bouts for estimation of CP [mean (SD); 213.3 (22.4) W] and W' [12.68 (3.08) kJ]. The subjects subsequently performed two variable-P tests to the limit of tolerance. During the first part, P was 117% or 134% of CP for a duration that expended approximately half of W'. The work rate was then abruptly increased to 134% (UP protocol) or decreased to 117% (DOWN protocol) of CP for the second part. RESULTS: There were no significant differences between W' [12.68 (3.08) kJ] and the total amount of work above CP during the UP [12.14 (4.18) kJ] and DOWN [12.72 (4.05) kJ] protocols (P > 0.05). CONCLUSION: We conclude that the work equivalent of W' is not affected by power variations during exhausting cycle ergometry, at least in the P range of 100-134% of CP. PMID- 12900699 TI - Age-related differences in 100-m sprint performance in male and female master runners. AB - PURPOSE: This study was undertaken to investigate age-related differences in the velocity and selected stride parameters in male and female master sprinters and to determine which stride characteristics were related to the overall decline in the performance of the 100 m with age. METHODS: The performances of 70 finalists (males 40-88 yr, females 35-87 yr) at the European Veterans Athletics Championships were recorded using two high-speed cameras (200 Hz) with a panning video technique and distance markers at 10-m intervals. Velocity, stride length (SL), stride rate (SR), ground contact time (CT), and flight time (FT) during the acceleration, peak velocity, and deceleration phases of the 100-m race were determined from the video records with the aid of the Peak Performance analysis system. RESULTS: There was a general decline in sprint performances with age, the decrease becoming more evident around 65-70 yr of age. The velocity during the different phases of the run declined on average from 5 to 6% per decade in males and from 5 to 7% per decade in females. Similarly, SL showed clear reductions with increasing age, whereas SR remained unchanged until the oldest age groups in both genders. Furthermore, the CT, which correlated with velocity, was significantly longer, and FT, which correlated with both velocity and SL, was shorter in older age groups. CONCLUSION: Our findings indicated that age associated differences in velocity in elite master sprinters were similar in each phase of the 100-m run. The deterioration of the overall performance with age was primarily related to reduction in SL and increase in CT. PMID- 12900700 TI - Cardiovascular response of trained preadolescent boys to mental challenge. AB - PURPOSE: To compare the cardiovascular reactivity response to mental challenge of aerobically trained and untrained preadolescent boys. METHODS: Heart rate (HR) and blood pressure, as well as stroke volume, total peripheral resistance, rate pressure product, and cardiac contractility response in 15 aerobically trained (10.3 yr) and 15 untrained (9.5 yr) subjects during and recovering from the Stroop task and a computer game (Tetris) were examined. Cardiac vagal activity was also assessed using time series analysis of heart period variability (HPV(ts)). RESULTS: Untrained compared with trained subjects made significantly more errors during Stroop, whereas performance on Tetris was similar. Trained subjects had significantly lower (P < 0.05) HR and significantly greater cardiac vagal activity at rest. Both groups had significant increases in HR, peripheral resistance, systolic and diastolic blood pressures, rate pressure product, and a significant decrease in vagal activity during Stroop. In contrast, only diastolic blood pressure increased and vagal activity decreased during Tetris. Cardiac contractility of both groups failed to increase during either stressor. Trained compared with untrained subjects had significantly lower absolute HR during and recovering from Stroop. Also HR and vagal activity change, relative to baseline, was significantly greater for the trained during Stroop. During and recovering from Tetris, absolute HR was significantly less for the trained subjects, but change scores were not significantly different. CONCLUSIONS: The greater HR and vagal activity change of trained subjects to the Stroop suggests greater cardiac reactivity to mental challenge. PMID- 12900701 TI - Short-term peak power changes in adolescents of similar anthropometric characteristics. AB - PURPOSE: The present study was undertaken to examine changes of cycling peak power (P(max)), optimal pedaling frequency (Vopt), and optimal pedaling force (Fopt) with age in subjects with the same lean leg volume (LLV), leg length (LL), and percentage body fat (%BF). METHOD: A total of 132 males aged 9.5-16.5 volunteered for this study. The population was divided into prepubertal (G1), pubertal (G2), and postpubertal (G3) groups. Within G1, G2, and G3, although the subjects were divided into three different age subgroups, there were no significant differences for LLV, %BF, and LL. RESULTS: Results showed that within G1, G2, and G3, P(max) increased significantly with age. Optimal velocity (Vopt) increased significantly with age in G1, whereas optimal force (Fopt) increased significantly with age into the other groups (G2 and G3). CONCLUSION: This study demonstrated that when anthropometric characteristics were controlled (LLV, LL, and %BF), P(max) and its two components (Vopt and Fopt) still increased with age. This indicates that other factors of qualitative nature have to be considered when determining P(max), Vopt, and Fopt. PMID- 12900702 TI - Measuring energy expenditure in habitually active and sedentary pregnant women. AB - PURPOSE: To describe patterns of energy expenditure (EE) during pregnancy and to assess the convergent validity of three methods of estimating EE. METHODS: We administered heart rate (HR) telemetry, accelerometry, and a physical activity record (PAR) over two consecutive days at weeks 20 and 32 of pregnancy and 12 wk postpartum to 28 habitually active and 28 habitually sedentary women. RESULTS: Mean daily waking-time EE at 20 wk by HR telemetry was 1814 (SD 443) kcal in active women and 1738 (448) kcal in sedentary women (P > 0.50), and did not change over the period of study (for active women P > 0.40; for sedentary women P > 0.70). Compared with HR telemetry, accelerometry underestimated EE by approximately 400 kcal x d-1, and the PAR overestimated EE by a similar amount, at all time periods in both active and sedentary women. EE, expressed per unit body weight, was consistently higher for active than for sedentary women during pregnancy. Pairwise correlations between methods ranged from 0.37 to 0.90 across time periods in both active and sedentary women. Correlations were lower (range 0.07-0.81) when adjusted for the length of the recording day. CONCLUSIONS: All methods were sensitive to variation in both the rate of EE and the duration over which activity was monitored. Accelerometry and PAR are useful methods for categorizing EE in epidemiologic studies among pregnant women but absolute estimates are biased relative to HR. PMID- 12900703 TI - Reexamination of validity and reliability of the CSA monitor in walking and running. AB - PURPOSE: To evaluate the reliability and validity of the CSA (model 7164) accelerometer (MTI) in a wide walking-running speed range in laboratory and field. METHOD: Twelve male subjects performed three treadmill walking/running sessions and one field trial with the same continuous protocol involving progressively increasing velocities at 5 min per interval from 3 to 6 km x h(-1) (walking) and 8 to 20 km x h(-1) (running). In the field trial, this protocol was terminated after 35 min (14 km.h(-1)), but the trial then extended with 5-km running at a freely chosen velocity. In both scenarios, two CSAs were mounted on each hip and the step frequency measured at each velocity. Oxygen uptake VO(2) x kg(-1) was measured on the last two treadmill sessions. Correlation analyses were performed for mean CSA output relationship with speed, VO(2) per kilogram, and step frequency. RESULTS: In all trials, CSA output rose linearly (R2 = 0.92, P < 0.001) with increasing speed until 9 km.h-1 but remained at approximately 10000 counts.min-1 during running, thus underestimating VO(2) per kilogram at speeds > 9 km x h(-1). Estimation errors increased with speed from 11% (P < 0.01) at 10 km x h(-1) to 48% (P < 0.001) at 16 km x h(-1), when assuming a linear relationship. Freely chosen velocities in the field trial ranged from 10.9 to 16.3 km.h-1. No difference in the CSA-speed relationship was observed between the two scenarios. Differences in CSA output between subjects could partially be attributed to differences in step frequency (R = -0.34 (P = 0.02) for walking and R = -0.63 (P < 0.001) for running). CONCLUSION: CSA output increases linearly with speed in the walking range but not in running, presumably due to relatively constant vertical acceleration in running. Between-subject reliability was related to step frequency because CSA data are filtered most at higher movement frequencies. Epidemiological CSA data should thus be interpreted with these limitations in mind. PMID- 12900704 TI - Validity of 10 electronic pedometers for measuring steps, distance, and energy cost. AB - PURPOSE: This study examined the effects of walking speed on the accuracy and reliability of 10 pedometers: Yamasa Skeletone (SK), Sportline 330 (SL330) and 345 (SL345), Omron (OM), Yamax Digiwalker SW-701 (DW), Kenz Lifecorder (KZ), New Lifestyles 2000 (NL), Oregon Scientific (OR), Freestyle Pacer Pro (FR), and Walk4Life LS 2525 (WL). METHODS: Ten subjects (33 +/- 12 yr) walked on a treadmill at various speeds (54, 67, 80, 94, and 107 m x min-1) for 5-min stages. Simultaneously, an investigator determined steps by a hand counter and energy expenditure (kcal) by indirect calorimetry. Each brand was measured on the right and left sides. RESULTS: Correlation coefficients between right and left sides exceeded 0.81 for all pedometers except OR (0.76) and SL345 (0.57). Most pedometers underestimated steps at 54 m x min-1, but accuracy for step counting improved at faster speeds. At 80 m x min-1 and above, six models (SK, OM, DW, KZ, NL, and WL) gave mean values that were within +/- 1% of actual steps. Six pedometers displayed the distance traveled. Most of them estimated mean distance to within +/- 10% at 80 m x min-1 but overestimated distance at slower speeds and underestimated distance at faster speeds. Eight pedometers displayed kilocalories, but except for KZ and NL, it is unclear whether this should reflect net or gross kilocalories. If one assumes they display net kilocalories, the general trend was an overestimation of kilocalories at every speed. If one assumes they display gross kilocalorie, then seven of the eight pedometers were accurate to within +/-30% at all speeds. CONCLUSION: In general, pedometers are most accurate for assessing steps, less accurate for assessing distance, and even less accurate for assessing kilocalories. PMID- 12900705 TI - Infant formulas. Recent developments and new issues. AB - Infant formulas on the market today should be aimed at providing the best alternative to breast milk for infants of those women who are unable to continue breastfeeding until 6 months of age and substituting ideally for human milk after 6 months of age approaching the structural and functional effects observed in breastfed infants. The aim is to mimic the functional outcome of the breastfed infant (e.g. growth and development), and not to copy the composition of human milk. For this purpose, the following compounds have been added to formulas and are reviewed: long-chain polyunsaturated fatty acids (LCPUFA) for brain composition and neurodevelopment, probiotics and prebiotics for the fecal flora and the local intestinal defense, and nucleotides for promoting the immune response. Changes in protein quantity and quality allow to balance the blood amino acid pattern (possibly relevant to the early stages of brain development for the neurotransmitter function) and reducing the protein intake could be important for the prevention of later overweight. Hydrolysed proteins are important in the prevention of atopic disorders. Many trials have been published so far with short-term assessments, most of them with positive findings. However, we need more data on the long-term follow-up of infants who were fed the new formulas. Such data will allow to look at neural performance, prevention of overweight and obesity, and effects on the immune-allergic pattern. PMID- 12900706 TI - Human milk substitutes. An American perspective. AB - Formula is a substitute for the preferred nutritional fluid for infants, human milk. Considering that human milk is uniquely suited for human infants, formula feeds have done remarkably well in promoting growth and development of bottle-fed and partially breast-fed infants. Early in the last century, the horrendously high infant mortality rate spurred the development of infant formula. A clean source of milk was thought to be necessary to overcome the mainly diarrheal deaths due to unhygienic conditions. Initially, the success of a formula was measured merely by the survival of the infant. As formulas more closely mimicked the performance of human milk, feedings were compared by the ability to support growth. Now, the additions and alterations to formulas have become more sophisticated and the outcome measures may be protection from infections, visual acuity and mental development. Studies to test these outcomes are difficult and often long term. Additionally, mere growth is recognized as an insufficient measure, rather quality of growth needs to be considered. Formula, like no other food, is potentially the sole source of an individual's nutrition for 4 to 6 months of life. In recognition of its importance, formula is the only food that is regulated by its own law in the United States, the Infant Formula Act of 1986. For the future, breast feeding will remain preferable for almost all infants and breastfeeding should be encouraged. Formulas will continue to be used and will continue to be needed. Improving and assuring the adequacy and safety of infant formulas is an increasingly difficult challenge. PMID- 12900707 TI - Formula feeding during cow's milk allergy. AB - Cow's milk allergy (CMA) is an immunologically mediated reaction to cow's milk proteins, which affects infant and young children. Cow milk elimination requires either breast-feeding, with or without elimination diet in mother or the use of specific formulas, based on cow's milk protein extensively hydrolyzed, which fit 90-95% of children with cow's milk allergy. In others, still reactive to allergic remnants in hydrolysates, an amino acid based formula is the optimal option. The good tolerance of soy formulas in a reasonable proportion of children with cow's milk allergy make them useful, including as a 1(st)-choice alternative, except probably for those below the age of 6 months. Any elimination diet in children is at risk of nutritional deficiency so that a constant monitoring of the growth parameters should be kept in those children. PMID- 12900708 TI - Formula-fed preterm neonates. AB - In very immature babies, nutrition often begined with human milk, but a lot of mothers have a difficulty in lactating and banked human milk is not always available. Therefore, preterm formulas have been specifically designed for very low birth weight (VLBW) infants during hospitalisation stay. They differ significantly from standard term formulas and their derivatives, such as extensively hydrolyzed protein formulas, which are not nutritionally adapted for these infants. Partially hydrolyzed protein formulas have been tried, but infants fed these formulas have a decreased nitrogen intestinal absorption rate. As a result the nitrogen content needs to be increased each time a partially hydrolyzed protein source is used in preterm formulas instead of an entire protein source. Although mineral retention in VLBW infants fed a formula is lower than in utero it might be sufficient. We recently observed an early catch up of bone mineralisation at theoretical term in VLBW infants fed a preterm formula containing highly soluble calcium salts. Probiotics and prebiotics are not yet well evaluated in preterm infants but might help in improving the development of physiologic intestinal flora and enteral feeding tolerance. There is still debate about optimal posthospital nutrition in preterm infants who are not breastfed. Feeding a nutrient-enriched formula provides a growth benefit when compared to infants fed a term formula, mainly during the first 2 to 3 months after discharge. Data about the harmful effects of using such formulas for a longer period are scarce. Therefore, the use of nutrient-enriched formula might be suggested for the first 2 months after discharge, as it is a crucial period in the development of these VLBW infants. PMID- 12900709 TI - Zinc absorption by infants. AB - Zinc is a vital mineral in human nutrition and rare cases of overt zinc deficiency are well described in term and preterm infants. A variety of methods have been developed to assess zinc absorption, retention and balance in humans, either using mass (metabolic) balance or stable isotope-based METHODS: The different methods have different benefits and disadvantageous. Many studies have used such methods to examine zinc absorption in term and preterm infants. In preterm infants the main interest has been establishing whether different diets lead to positive zinc balance and approached in utero accretion rates. The earliest studies were concerning as they suggested that preterm infants may be in negative zinc balances for prolonged periods after birth. Subsequent studies have been more contradictory and it remains unclear from the literature how much zinc preterm infants absorb early in life. Nor is it clear whether poor zinc absorption during this period has any long-term consequences. Studies in term infants have suggested no such prolonged period of negative zinc balance. Generally the absorption values are in line with those used by the Institute of Medicine in the most recent edition of the Dietary Reference Intakes. Other studies have been designed to assess the effect of changes in formula composition on zinc intake (e.g. the presence or absence of lactose, high vs low zinc contents, etc.). Ho-wever, despite almost 20 studies of zinc absorption in infants many physiologically important questions remain unanswered, especially in preterm infants. PMID- 12900710 TI - Feeding disorders in infancy: an empirical study on mother-infant interactions. AB - AIM: The objective of this paper is an empirical analysis of mother-infant feeding interactions through an observational instrument (Feeding Scale), which is applied to the videotape of a feeding session. METHODS: Three samples of mothers and their children have been examined (104 couples with children ranging in age from 2 to 36 months: a first group in which children presented a feeding disorder on organic and functional base (food intolerances and/or gastroesophageal reflux), a second group in which the disorder was not due to a medical condition and a third group of control. The Authors meant to test three hypotheses: 1) the presence of developmental changes in mother-infant interactional patterns during the course of the first three years of life; 2) higher degree of dysfunction of mother-infant interaction in those dyads in which the feeding disorder had not organic causes; 3) dysfunctional interactions in the couples of mothers and children in which the feeding disorder was on an organic base. RESULTS: The results of the statistic analyses have confirmed the hypotheses, providing different elements of reflection. CONCLUSIONS: The Authors underline the importance of a multidimensional, global and integrated approach in the clinical assessment and treatment, which evaluates the possible interconnections among organic, psychogenetic and relational factors in the origin and course of feeding disorders in infancy and early childhood. PMID- 12900711 TI - [Psychological findings in children with short stature]. AB - AIM: The aim of this study is to evaluate the psychological findings in patients with short stature. METHODS: We studied 19 subjects, 13 males and 6 females, with age range 7-14 years. We evaluated heigth, growth velocity, bone age, target height and growth hormone secretion after provocative stimuli. Psychological evaluation included: Kovacs Scale, Children's Depression Inventory (CDI), Anxiety Scale (Busnelli-Dall'Aglio-Farina); drawing of the human figure (Goodenough Test); Raven Test for neuropsychological performances (P.M. 38 and 47). Statistical analysis was performed using Mann-Whitney U-test. RESULTS: We diagnosed familial short stature (FSS) in 7 patients and growth hormone deficit (GHD) in 12. No statistical difference was found in the anxiety and depression tests, although the score was higher in GHD patients. The human figure drawing and the interview revealed low self-esteem, sense of inadequacy, dependence from parents, social inhibition in all patients. These characteristics were more evident in patients with GHD. Neuropsycho-logical evaluation by Raven test showed normal score in all patients, however subjects with FSS exhibited a higher score than with GHD (p<0.05). CONCLUSIONS: Our data suggest a negative influence of short stature on the affective field of children with short stature; GHD patients exhibited lower neuropsychological performances and more psychological problems than patients with FSS. PMID- 12900712 TI - Psychopathological assessment in children affected by Duchenne de Boulogne muscular dystrophy. AB - BACKGROUND: Several studies on the intelligence of children suffering from Duchenne muscular dystrophy (DMD) found impairments in linguistic functions, memory and affective development. Memory deficit mainly involves long-term memory; short-term memory impairment was also present, even if less frequently. Some studies tried to establish a correlation between the distal deletion of the dystrophin gene and low intelligence levels. METHODS: The present study investigated the cognitive, emotional and relational states in a group of DMD children. The patients were selected on the basis of physical examination, muscle biopsy, histochemical analysis, genetic testing, regular schooling. Molecular genetic testing detected a deletion in 2 high frequency deletion regions in all subjects. The following psychodiagnostic tests were used for assessment: WISC-R, Spontaneous Drawing, Draw-A-Person test, Family Drawing, Thematic Apperception test and Rorschach test. RESULTS: No impairment of the intellectual function was detected in the group of children we examined; only 2 subjects had a total IQ below average. Instead, psychic disorders were detected such as proneness to isolation, self-depreciation, past experiences of marginalization, minor depressive symptoms, insecurity signs, hypochondriac worries and high level of anxiety. In addition children exhibited poor adaptation to the environment and labile affectivity, scarcely controlled by the higher conscious levels. CONCLUSIONS: These disorders are often a dynamic prolongation of a psychological process starting when the diagnosis is made and continuing in a slow and latent fashion throughout the evolution of the disease. PMID- 12900713 TI - [Human recombinant tissue-transglutaminase antibodies: evaluation of a commercial kit and comparison of methods using guinea pig antigen]. AB - AIM: The tissue-Trans-Glutaminase (t-TG) has been reported to be the target for endomysial antibodies in coeliac disease. Coeliac disease if untreated, although clinically silent, predispose for other autoimmune disease, large scale screening methods are needed for an early diagnosis. The recently introduced ELISA methods to detect tTG antibodies, that used as antigen Guinea Pig tissue transglutaminase provide an efficient alternative to the anti-endomysial (EMA) immunofluorescent method and are suitable for screening. Our aim was to compare the commercial kit utilizing tTG from guinea pig (GP-tTG) to the standard method that we developed in our laboratory and with the new method that employed human recombinant tTG (hr tTG). METHODS: We tested serum samples from 16 untreated celiac patients, 10 coeliac patients in gluten free diet, 22 subjects with other disease and 32 healthy controls, for a total number of 80 sera, with 3 commercial kit which employed GP-tTG and 1 that employed hr-tTG, as reference method we used an ELISA method developed in our laboratory and the immunofluorescence method to detect EMA. RESULTS: Results show that methods employing human tTG had a sensibility of 100% with a specificity of 98% and a predictive value of 94%, instead methods employing GP-tTG show inferior results. CONCLUSION. We conclude that is better using hr-tTG as antigen in screening methods for coeliac disease. PMID- 12900714 TI - Delayed presentation of congenital diaphragmatic Bochdalek hernia. Case report. AB - The authors describe the case of a 1-month-old patient admitted for crisis of paleness, accompanied by hyporeactivity, tachycardia and polypnea; the symptoms, attenuated after detension of the abdomen, grew worse, with compromise of general conditions and respiratory failure. Chest X-ray shows the herniation of intestinal loops in the thoracic cavity. After the first emergency care, it became necessary to submit the baby to HFO and cardiovascular drugs for some hours before getting oxygenation index (OI) and circulation stabilization. Late presenting congenital diaphragmatic hernia (LCDH) is a condition that can occur during first or second infancy with respiratory or abdominal symptoms, quickly worsening or intermittent. The prognosis is good if diagnosis is timely. Patient stabilization before surgical intervention must be valued not only by O.I. but also by circulation conditions. PMID- 12900715 TI - Spontaneous intestinal perforation in a full-term infant: association with infection. AB - The term spontaneous intestinal perforation suggests a perforation in the gastrointestinal tract of a newborn of no demonstrable cause. Only a few cases have been described in full-term newborns. The aetiology and pathogenesis of the disease are unknown although multiple theories have been proposed. Some authors suggest ischemia as the most likely cause. Conditions associated with fetal or neonatal hypoxia are important antecedents for this emerging distinct entity. We present a case of a spontaneous, intestinal perforation in a full-term neonate with urinary tract infection. There was no clinical evidence of necrotizing enterocolitis or bowel obstruction. Radiological images revealed a pneumoperitoneum. An emergency explorative laparotomy was performed. A localized linear perforation was identified in the transverse colon. Pathological examination of the resected specimens failed to reveal any etiology for the perforation. The neonate recovered rapidly, with no gastrointestinal complications. In our case none of the factors which have previously been associated with intestinal perforation could be implicated. We suggest that focal intestinal perforation is possibly the result of infection. Further studies, including careful recording of cases and close histopathological examination of resected specimens, are required in order to provide more information and improve our understanding of the aetiology of this rare occurance. PMID- 12900716 TI - Spontaneous pneumomediastinum: is a chest X-ray sufficient? AB - Spontaneous pneumomediastinum (SPM) is an uncommon disease defined as a non traumatic presence of free air in the mediastinum, without underlying disease. We present a 13-year- old boy who was previously in a perfect health, who was presented with subcutaneous cervical emphysema, dysphagia, chest and neck pain. The chest roentgenogram revealed the presence of subcutaneous emphysema without any other abnormal findings. A computer tomography (CT) scan was obtained, and this confirmed the existence of subcutaneous cervical emphysema and also the presence of pneumomediastinum. The child's progress was uneventful and after 6 days he was discharged from the hospital in excellent clinical condition. We propose that chest CT is useful, in less obvious cases of SPM, to detect the free air in the mediastinum and probably SPM is underdiagnosed in clinical practice in the young people. PMID- 12900717 TI - Organic nitrate tolerance and endothelial dysfunction: role of folate therapy. AB - Tolerance to organic nitrates has been demonstrated in patients with acute coronary syndromes following continuous, long-term therapy, and has been shown to occur within 24 to 48 h after administration of a nitrate preparation. Dosing schedules that include a nitrate-free period often fail to ameliorate the development of nitrate tolerance, and, in fact, can result in an increase in rebound ischemia. Several mechanisms have been suggested to explain the phenomenon of nitrate tolerance, notably, nitrate-mediated depletion of intracellular thiols, and enhanced reactive oxygen species formation. Recently, increased superoxide production, owing to the un-coupling of the endothelial isoform of nitric oxide synthase (eNOS) and/or increased NAD(P)H oxidase activity, has been implicated in the development of nitrate tolerance. Based on these observations, strategies to overcome tachyphylaxis to nitrates have been designed to modulate the production of reactive oxygen species. Folic acid and its derivatives have been shown to prevent nitrate tolerance by preventing eNOS uncoupling, and, thereby, eNOS-mediated superoxide production resulting in improved endothelial function. Folic acid, which has a benign side-effect profile, may, therefore, be a simple pharmacological intervention to prevent nitrate tolerance and may have broad application in the treatment of atherothrombotic vascular disease. PMID- 12900718 TI - [Therapeutic indications for acute venous thromboembolism. Current status and future perspectives]. AB - Except for selected patients requiring aggressive therapies, the large majority of patients with acute venous thromboembolism are currently treated with full doses of unfractionated or low-molecular-weight heparins (LMWH) followed by oral anticoagulants for variable periods of time. LMWHs present a number of potential advantages over unfractionated heparin: a longer plasma half-life, improved subcutaneous bioavailability, and a more predictable dose-response relationship. As a result of these pharmacokinetic properties, these compounds have the potential to greatly simplify the initial treatment of venous thromboembolism, making the treatment of suitable patients feasible in an outpatient setting with considerable saving in costs and improvement in patients' quality of life. The use of unfractionated heparin is still desirable in the initial treatment of acute pulmonary embolism in non-critically ill patients. The use of heparin protocols assures that virtually all patients will promptly achieve the therapeutic range for the activated partial thromboplastin time. Although the optimal duration of anticoagulation in patients suffering an episode of venous thromboembolism is presently unknown, it seems reasonable to administer a short term course of coumarin drugs to patients with thrombosis associated with transient risk factors, while a longer course should be considered in patients with idiopathic thrombosis and in those with permanent risk factors. At present, indefinite anticoagulant therapy remains a clinical judgment in the individual patient. The efficacy and safety of emerging drugs (pentasaccharide, ximelagatran) in the treatment and secondary prevention of venous thromboembolic disorders is currently under investigation. PMID- 12900720 TI - [Early effects of cigarette smoking in hypertensive and normotensive subjects. An ambulatory blood pressure and thermographic study]. AB - BACKGROUND: Among the acute effects of cigarette smoking there are an increase in the arterial blood pressure and a decrease in skin temperature. The aim of this study is to evaluate whether these effects are more evident in hypertensive subjects (HTS) than in normotensive (NTS) ones. METHODS: A monitored Ambulatory Blood Pressure method, an Agema 880 Thermograph and a Surgitron thermal stimulator have been employed. Ten NTS males and 10 HTS, age paired (50-60 years, mean 57.8), were examined. Area of interest, the volar face of the fingertips. The pressural and thermographic tests were performed with patients staying in a proper room for 2 h, smoking consecutively 4 cigarettes, one every 14 min. The tests were made at 0 time (baseline), after smoking each cigarette and, finally, 60' after the beginning of the last one. RESULTS: A rise in blood pressure and a fall in skin thermal gradients both in HTS and in NTS was seen since the first cigarette, and these values, more evidently in hypertensive subjects, were appeared progressively increased along with the other 3 cigarettes. CONCLUSIONS: These results confirm the damage of smoking on peripheral blood vessels verified and also the cumulative effect of smoking more cigarettes. The similar increased percentage of ABP and telethermography (TT) values in these tests lead us to consider TT as a routinary method for the evaluation of a more general vascular damage provoked by cigarette smoking. Also, the immediate visual thermal effect (=thermal amputation) after smoking is very useful in smoking educative programs. PMID- 12900719 TI - Multidetector helical angio CT oblique reconstructions orthogonal to internal carotid artery for preoperative evaluation of stenosis. A prospective study of comparison with color Doppler US, digital subtraction angiography and intraoperative data. AB - AIM: The recent introduction to clinical practice of multidetector helical angio CT (MHACT) has generated a new interest in the diagnosis of carotid artery atherosclerosis. In recent years there has been a redefinition of the indications to carotid artery endarterectomy; there is a tendency to appraise plaque morphology and composition, and not only stenosis degree. The aim of this prospective study is to analyze the validity of MHACT in the diagnosis of atheromasic stenosis of the carotid bifurcation, in comparison with US color Doppler (USCD), digital subtraction angiography (DSA) and intraoperative evidence (OP). Special emphasis is given to the analysis of plaque composition and to precise evaluation of the stenosis percentage computed as an area rate on oblique reconstructions performed exactly orthogonal to the axis of the vessel at the point of maximal stenosis. METHODS: Twenty-seven carotid stenosis (in 24 patients) were preoperatively evaluated by USCD, MHACT, DSA. We calculated the stenosis degree with USCD, by the ECST method, both as a diameter rate and as a bidimensional value. By MHACT we computed the percentage of stenosis as an area rate, in an MPR oblique plane orthogonal to the vessel axis, at the point of maximal stenosis; by DSA we used NASCET and the common carotid artery method (CC). DSA measurements have been mathematically converted as area rate (NASCET2 and CC2). All the patients were operated on with the eversion technique (EEA); it was always possible to obtain an intact cylindrical specimen of the plaque and to perform a reliable and accurate evaluation of the degree of stenosis by sectioning and measuring it on the desk. We gave a score to the presence and amount of lipid and calcium components, and these data were compared to those obtained with the different diagnostic instruments. RESULTS: Statistical analysis of the 7 data sets showed an important underestimate of the angiographical method (DSA), even if the measurements were carried on by a precision caliper. The mathematical conversion of a linear stenosis rate into an area rate gives these values more reliability, indicating, as for NASCET2, non statistically significant differences when compared to surgical evidence, notwithstanding a high standard deviation. The rates obtained by USCD (area rate) and MHACT proved to be very accurate with respect to surgical evidence. In this study there was no significant difference in the detection of various components (lipid and calcium) of the atheromasic lesion, by MHACT and surgical evidence (OP). CONCLUSION: MHACT promises to be a very accurate instrument in the detection of plaque components and identification of the stenosis degree. It shows some limits in very calcific lesions and occasionally, in relevant cardiac failure, may be useless. Hence one feels the need to reassess the role of DSA, once considered the "gold standard", with more extensive prospective studies, including comparison with MHACT and USCD. PMID- 12900721 TI - A new autologous venous valve by intimal flap. One case report. AB - Various surgical techniques have been proposed for the treatment of chronic venous insufficiency of post-thrombotic recanalized deep veins of the lower limbs. The preferable method seems to be represented by intravenous valvuloplasty except for the cases affected by extensive valvular damage. For this reason some experimental autologous, heterologous and prosthetic venous valves have been proposed. Such a problem emerged for 1 patient (male, aged 78 years, right limb, leg dystrophy, multiple ulcerations at the ankle) which was selected by duplex, Doppler venous pressure index, photoplethysmography and ascending phlebography. An iliac-femoral and popliteal post-thrombotic, recanalized, decompensated venous insufficiency and one Cockett's perforator incompetence were diagnosed (CEAP classification: C6s Es As2d14 Pr). A bicuspid apparently repairable popliteal valve was detected by phlebography. A traditional intravenous valvuloplasty was planned but the valve was not found at surgical exploration. A monocuspid valve reconstruction by intimal flap vein was performed. The following results were obtained and controlled after one year: stable ulceration healing, dystrophy reduction, improvement in the quality of life, normalization of the hemodynamic parameters and of the radiological morphology of the new valve. It can be concluded that monocuspid valvular repair by intimal flap can be successfully performed in cases affected by secondary valveless deep venous insufficiency of the lower limbs. PMID- 12900722 TI - [Treatment of a femoral venous aneurysm with foam-sclerotherapy]. AB - Venous aneurysms are uncommon. Venous dilatation and large varices could be treated with injections of foam and sclerosing agents followed by local compression which obliterate or thrombose the aneurysmal space. However, the use of foam-sclerotherapy to obliterate venous aneurysms has never been reported before. The obstruction or sclerosis of the aneurysmal space should be obtained without altering the femoral flow. The patient is a 65-year old lady without history of trauma involving the femoral region. A compressible lump is present at the third, internal part of the left femoral fold. The lump is painless and easily obliterated by compression. Color duplex shows a large cavity (4 x 4 x 6 cm) and venous flow is visible. Flow produces a venous jet, visible with color and ejecting from the common femoral vein. A first injection is made under ultrasound guide using a foaming agent: 10 ml of foam including 4 ml of 3% sclerosing solution are injected. During the injection the femoral vein is compressed with the probe to avoid passage of the sclerosing foam into the femoral vein. After 2 weeks, ultrasound indicate only a partial occlusion of the aneurysm. The procedure is repeated and after 2 more weeks the ultrasound scan shows a complete occlusion of the aneurysm. The femoral vein and the long saphenous vein are patent. After 12 weeks the situation is unchanged and the results appear to be permanent. This new method, never described before is an important, minimally invasive method in case of venous aneurysms. Results from larger studies should be available to define indications and modalities of treatment. PMID- 12900723 TI - [Chronic venous insufficiency: an open trial of FLEBS Crema]. AB - AIM: This study was realized to evaluate the FLEBS CREMA tolerability and its therapeutic efficacy, in patients with functional disease linked to not complicated venous or lymphatic insufficiency. This compound acts as vasoconstrictor thanks to the vegetable extract of Ruscus aculeatus, but it is also able to reduce the edema, thanks to the vegetable extract of Melilotus officinalis. METHODS: All the patients were treated for three weeks with FLEBS CREMA, that was applied 2 or 3 times a day, on both legs. The traditional clinical parameters of chronic venous insufficiency (IVC) (edema, pain, heaviness, itch, cramps and other symptoms) were evaluated during the treatment. RESULTS: The results show an improvement for all the monitored symptoms, also if the positive clinical response was different for each evaluated parameter. The best results were obtained for edema, pain, heaviness and itch: the statistical analysis shows indeed a significant difference between the beginning and the end of the treatment. In this study the dermo-cosmetic qualities and the tolerability of FLEBS CREMA were also evaluated: both these parameters obtained a positive clinical evaluation. CONCLUSIONS: Basing on these data, the final evaluation for this compound was good/very good for the 80-90% both doctor and patients. PMID- 12900724 TI - Cardiac tamponade mimicking ventricular rupture after thrombolysis for acute myocardial infarction. PMID- 12900725 TI - Trends in movement disorders' therapy. PMID- 12900726 TI - Cell transplantation for Parkinson's disease in the age of deep brain stimulation. PMID- 12900727 TI - Neural stem cells. Biological features and therapeutic potential in Parkinson's disease. AB - AIM: Neural stem cells (NSC) are clonogenic cells, capable of self-renewal and multilineage differentiation, since, under the appropriated experimental conditions, they proliferate indefinitely as undifferentiated neurospheres or differentiate in neurons, astrocytes and oligodendrocytes. Parkinson's disease is a neurodegenerative disorder characterized by the progressive loss of dopaminergic neurons. METHODS: Here we investigated the suitability of recently identified and characterized neuronal progenitor cells at eliciting functional recovery in unilateral 6HODA-lesioned mice. We describe herein that intrastriatal engraftment of stem cell-derived neurons isolated from the olfactory bulb to give rise dopaminergic-like neurons results in long lasting functional recovery in 6OHDA-injured mice. RESULTS: Unilateral injection of 6OHDA resulted in a progressive neurodegeneration of the nigro-striatal pathway. Likewise, the systemic administration of L-DOPA in these mice elicited a marked contralateral turning which was evident 1 week post, increased during the following week and than stabilize throughout the time of the experiment. Conversely, the intrastriatal implantation of partially differentiated stem cells at 14 days postlesion, resulted in a profound decrease in L-DOPA-induced circling behavior; interestingly, the effect was evident 1 week after the engraftment and was retained during the following 9 weeks. Detailed biochemical and immunohistochemical evaluation is currently under investigation in our laboratory. Conclusion. Our observation opens new perspectives for the treatment of neurodegeneration in Parkinson's disease. PMID- 12900728 TI - Different methods for anatomical targeting. AB - AIM: Several procedures are used in the different neurosurgical centers in order to perform stereotactic surgery for movement disorders. At the moment no procedure can really be considered superior to the other. We contribute with our experience of targeting method. METHODS: Ten patients were selected, in accordance to the guidelines for the treatment of Parkinson disease, and operated by several methods including pallidotomy, bilateral insertion of chronic deep brain electrodes within the internal pallidum and in the subthalamic nucleus (18 procedures). INTERVENTIONS: in each patient an MR scan was performed the day before surgery. Scans were performed axially parallel to the intercommissural line. The operating day a contrast CT scan was performed under stereotactic conditions. MEASURES: after digitalization of the MRI images, it was possible to visualize the surgical target and to relate it to parenchimal and vascular anatomic structures readable at the CT examination. The CT scan obtained was confronted with the MR previously performed, the geometrical relation between the different parenchimal and vascular structures and the selected targets were obtained. Stereotactic coordinates were obtained on the CT examination. RESULTS: It was possible to calculate the position of the subthalamic nucleus and of the internal pallidum on the CT scan, not only relating to the intercommissural line, but considering also the neurovascular structures displayed both on the MRI and the CT scans. CONCLUSION: The technique that our group presents consist in an integration between information derived from the CT and the MR techniques, so that we can benefit from the advantages of both methods and overcome the disadvantages. PMID- 12900729 TI - MRI indirect stereotactic targeting for deep brain stimulation in Parkinson's disease. AB - AIM: Stereotactic localisation of brain targets for functional neurosurgery might be obtained with different neuroimaging sources. Magnetic resonance (MR) might be of particular interest for its high definition of the various brain structures; unfortunately, magnetic field distortion and inaccuracy in imaging reproduction constrain the use of MRI in stereotactic surgery. METHODS: We present an experimental work devoted to the utilization of MRI in the targeting of the subthalamic nucleus for deep brain stimulation (DBS) in Parkinsonian patients. RESULTS: Experimental data were obtained after PVC phantom and fixed human brain measurements of the stereotactic coordinates of specific basal ganglia structures. Intrinsic and external sources of error and imaging distortion have been carefully corrected. Afterwards, 36 patients, operated for DBS in the subthalamus for Parkinson's disease (PD), have been investigated. MRI targeting proved to be clinically successful; comparing anatomical and neurophysiological findings, MRI targeting scattered from the neurophysiological targeting in a minority of cases. CONCLUSION: MRI targeting proved to be reliable for functional stereotactic surgery, pending careful and adequate quality control of the distortion and of the sources of error. PMID- 12900730 TI - Multiple sequential image-fusion and direct MRI localisation of the subthalamic nucleus for deep brain stimulation. AB - AIM: Deep brain stimulation (DBS) is the treatment of choice for advanced Parkinson's disease. The target co-ordinates are traditionally calculated in relation to the intercommissural distance. Anterior (AC) and posterior commissures (PC) may be visualised by the means of ventriculography, CT or MRI. METHODS: We have studied the efficacy of direct visualisation of the subthalamic red nucleus complex on MRI, the advantage of fusion of stereotactic CT and MR images (Multiple Sequences Image Fusion - MuSIF). These methods are combined with double check of indirect calculation of the target co-ordinates based on AC-PC line, as well as the corrispondence to the stereotactic electronic atlas. RESULTS: Subthalamic nucleus (STN) was well recognisable in fused images in all 22 sides. At 3 months from surgery it was possible to reduce 76% of L-dopa equivalent daily dose. Dyskine-sias reduced to 50% and motor fluctuation up to 45%. CONCLUSION: In our experience MuSIF offers very high rate of accuracy in calculation of target co-ordinates. Direct visualisation of STN in MR and MuSIF are reliable and facilitate the accuracy of identification of target co ordinates. Intraoperative neurophysiological recording increases the accuracy of microelectrode position. PMID- 12900731 TI - Functional neuroimaging (PET and SPECT) in the selection and assessment of patients with Parkinson's disease undergoing deep brain stimulation. AB - Deep brain Stimulation (DBS) is an effective treatment for patients with advanced Parkinson's disease (PD) and motor complications who can no longer be improved by adjustment of medical therapy. Selection of surgery candidates and follow-up after surgery are critical for good outcome. Functional neuroimaging can help in the clinical assessment of these patients. We have used single photon emission computed tomography (SPECT) and the tracer ECD to measure regional cerebral blood flow before and 6 months after DBS of the subthalamic nucleus (STN) in 20 patients with advanced PD. We found a significant increase in the anterior cingulate/supplementary motor cortex in the 12 good responders (change in off unified UPDRS >50%). Conversely, patients with poor response (n=8; change in off UPDRS-III <50% following DBS) revealed a significant worsening of cortical hypoperfusion particularly in the prefrontal areas. No flow decrements were detected in the basal ganglia and in the thalamus in both groups during DBS stimulation suggesting that DBS does not have a "lesion like" effect. If DBS stimulates and does not inactivate STN projection neurons, flow reduction in the poor responders may be secondary to increased inhibitory basal ganglia output. PMID- 12900732 TI - Deep brain stimulation in Parkinson's disease: bilateral implantation of globus pallidus and subthalamic nucleus. AB - AIM: Deep brain stimulation (DBS) of subthalamic nucleus (STN) and of the pars interna of globus pallidus (GPi) is used to improve Parkinsonian symptoms and attenuate levodopa-induced motor complications in Parkinson's disease (PD). What are the physiological effect of DBS and the best anatomical structure to stimulate are still not completely clear. In this way we could evaluate the clinical effects of simultaneous stimulation of STN and GPi as well as the isolated stimulation of each target. METHODS: The stereotactic methods used to localise STN and GPi were based on non-telemetric ventriculography, with 3P Maranello or Leksell Stereotactic System. The effects of DBS have been assessed in 13 cases of PD, immediately after (30 minutes) the stimulation has turned on and during chronic stimulation (weeks or months). RESULTS: Most of the studies have been conducted on patients with STN implantation, and these studies reported relevant improvement in motor function and relatively low rate of complication. CONCLUSION: The large experience of ablative surgery associate with the DBS experience of some group worldwide indicate that GPi is a possible and very promising target for the management of Parkinsonian symptoms. Our patients demonstrate in acute and chronic evaluation, the best clinical results with contemporary activation of DBS in both targets. PMID- 12900733 TI - Treatment of dystonic syndromes by chronic electrical stimulation of the internal globus pallidus. AB - AIM: Dystonia is a medically intractable condition causing twisting or myoclonic movements and abnormal postures. There is an important heterogeneity among etiologies of dystonia. The electrical stimulation of the globus pallidus has been used successfully in primary generalized dystonia. The aim of this study was to examine the long-term efficacy and safety of deep brain stimulation (DBS) in the treatment of primary and secondary generalized dystonia in children and adults. METHODS: Fifty-three patients were included. Electrodes were bilaterally implanted under stereotactic guidance and connected to neurostimulators, subcutaneously inserted. Efficacy was evaluated by comparing scores on the clinical and functional Burke-Marsden-Fahn dystonia rating scales (BMFDRS) before and after implantation. Patients were divided into 3 groups: group 1 comprised 15 patients with DYT1 dystonia; group 2, 17 patients with dystonia of unknown etiology and group 3, 21 patients with secondary dystonia. The mean follow-up was 26.6+/-12.3 months for primary dystonia and 23.1+/-11.8 for secondary dystonia. RESULTS: After 1 year, the improvement of the clinical score is 71% in group 1, 74% in group 2 and 31% in group 3. The functional score was improved by 63% in group 1, 49% in group 2 and 7% in group 3. We did not find any significant difference between children and adults. In secondary dystonia, efficacy of the stimulation is more limited. The efficacy of the stimulation improved with time for the 3 groups. COMCLUSION: Electrical stimulation of the internal globus pallidus proved to be an effective treatment for generalized dystonia and should be considered as first-line therapy. PMID- 12900734 TI - Do intraoperative microrecordings improve subthalamic nucleus targeting in stereotactic neurosurgery for Parkinson's disease? AB - AIM: The clinical importance of intraoperative microrecordings for subthalamic nucleus (STN) localization in neurosurgical practice remains a matter of debate in the various groups. METHODS: To investigate their usefulness in localizing the STN, we retrospectively evaluated how intraoperative microelectrode recordings changed the targeting of the STN estimated only on intraoperative stimulation and neuroanatomic targeting procedures. For neuroradiologic targeting of the nucleus we used a TC-MRI fusion algorithm and direct visualization of the STN. Besides standard microrecordings we also analyzed the power spectral density (PSD) pattern of physiological signals along the track and its neuroanatomic and clinical correlations. RESULTS: In our series of 12 patients with Parkinson's disease undergoing surgery for implantation of deep-brain stimulation (DBS) electrodes in the STN we found that in 25% (1/4) of patients, microrecordings determined the choice of the optimal track. In all the tracks analyzed the PSD peak coincided with the point selected for the final electrode implantation on the basis of the standard procedure for intraoperative monitoring based on both microstimulation and recordings. CONCLUSION: Intraoperative microrecordings are of determinant importance for accurate STN localization and are essential for optimal results in neurosurgical practice. PSD analysis is a simple and quick quantitative signal descriptor that will probably provide even more precise, simple and rapid tool for intraoperative neurophysiological localization of the STN. PMID- 12900735 TI - Regional brain injury epidemiology as the basis for planning brain injury treatment. PMID- 12900736 TI - ICNC VI and beyond. PMID- 12900738 TI - Effect of antianginal medication on resting myocardial perfusion and pharmacologically induced hyperemia. AB - BACKGROUND: Patients scheduled for myocardial perfusion imaging are often taking several antianginal drugs. There is presently no consensus concerning a regimen of discontinuation before either rest or pharmacologic stress myocardial perfusion imaging. Whether antianginal treatment affects diagnostic sensitivity and specificity is not well documented. Methods and results The effect of the three most commonly used antianginal drugs (nitroglycerin, 400 microg [NTG]; metoprolol, 50 mg [MET]; and amlodipine, 5 mg [AML]) on myocardial perfusion was tested in 49 patients (age, 63 +/- 8 years; 43 men) allocated prospectively to one of the treatments (NTG, n = 25; MET, n = 14; and AML, n = 10). All patients had documented coronary artery disease and were scheduled for elective percutaneous coronary intervention. Patients were studied once on treatment and once off treatment with an interval of 1 to 3 weeks. For NTG, the measurements were performed on the same day with an interval of 1 hour. The MET and AML groups were also studied during dipyridamole-induced hyperemia (0.56 mg. kg(-1). min(-1) for 4 minutes). So that a quantitative value of myocardial perfusion in milliliters per gram per minute could be obtained, myocardial perfusion was quantified with nitrogen 13 ammonia positron emission tomography as an average of the midventricular perfusion in each of the 3 vascular territories. NTG treatment increased the overall resting perfusion (0.75 +/- 0.18 vs 0.86 +/- 0.22, P <.05), whereas resting perfusion was reduced after MET treatment (0.92 +/- 0.14 vs 0.82 +/- 0.17, P <.05). AML treatment did not alter resting perfusion (0.87 +/- 0.22 vs 0.87 +/- 0.23, P = NS). Dipyridamole-induced hyperemia was reduced after treatment with MET (2.02 +/- 0.66 vs l.57 +/- 0.52, P <.001), whereas the hyperemic response was unchanged after treatment with AML (1.54 +/- 0.49 vs 1.86 +/- 0.91, P = NS). CONCLUSIONS: Antianginal medication can alter both resting and hyperemic myocardial perfusion and might affect the ability to detect flow limiting stenosis. NTG increases perfusion, MET reduces perfusion, and AML does not affect perfusion. Larger-scale trials are warranted to establish a consensus for optimal antianginal medication for patients undergoing perfusion imaging. PMID- 12900739 TI - Correct spatial normalization of myocardial perfusion SPECT improves detection of multivessel coronary artery disease. AB - BACKGROUND: This study sought to improve the detection of multivessel coronary artery disease (CAD) with the use of rest and exercise single photon emission computed tomography (SPECT) perfusion scintigraphy by developing a processing scheme, which provides proper regional normalization of the images for interpretation. When SPECT perfusion images are interpreted, one area of myocardium serves as "normal." We hypothesized that if this "normal" region changes location from rest to stress, the stress images must be adjusted for proper interpretation. By taking into account the level of tracer activity in this "normal" area on the resting images, we could more accurately identify patients with multivessel CAD. Methods and results Dual-isotope rest (thallium 201) and exercise dobutamine or adenosine stress (technetium 99m sestamibi) perfusion SPECT studies were examined in 258 patients with 2- or 3-vessel CAD on coronary arteriography performed within 6 months of each other (mean interval, 19 days). If a shift in regional location of the "normal" segment from rest to stress was present, the images were (1) interpreted in the usual fashion for the number of vessels with ischemia (PRE-NORM) and (2) reinterpreted after quantitative normalization (ie, adjusting the display window until the intensity of the "normal" segment was matched at rest and stress [POST-NORM]). Interpretation was performed with blinding to arteriographic results. An angiographic stenosis was defined as luminal diameter stenosis greater than 50%. Three control groups comprising (1) single-vessel CAD (n = 119), (2) no significant angiographic CAD (n = 118), and (3) a normalcy group of low pre- and post-test probability of CAD (n = 44) were also studied to determine the incidence of false-positive results induced by the renormalization technique. A shift in the "normal" segment occurred in 81 studies of 258 patients (31%), 80 of which were read as abnormal PRE-NORM (sensitivity, 99%); however, for their 216 stenosed vessels, only 143 were detected PRE-NORM (vessel sensitivity, 66%; accuracy, 65%). The mean POST-NORM change in the display was 11%. POST-NORM, all 82 patients' studies were interpreted as abnormal (sensitivity, 100%), and 196 of 216 vascular territories were abnormal (vessel sensitivity, 91%; accuracy, 83%; both P <.0001 vs PRE-NORM). In the single-vessel disease, no significant disease, and normalcy groups, 19 of 119, 15 of 118, and 11 of 44 patients, respectively, demonstrated a shift in the peak pixel location. However, there were no significant changes in single-vessel sensitivity, angiographic specificity, or normalcy in these patients. CONCLUSIONS: With multivessel CAD, the "normal" region on SPECT often changes in location from rest to stress, potentially masking the extent and severity of multivessel ischemia. Renormalization of the images to match their resting level before image interpretation allows diagnosis of contralateral ischemia and strikingly improves the detection of multivessel CAD, without a substantive loss in specificity. PMID- 12900740 TI - Residual exercise SPECT ischemia on treatment is a main determinant of outcome in patients with coronary artery disease treated medically at long-term with beta blockers. AB - BACKGROUND: Beta-blockers are potent anti-ischemic medications, able to improve prognosis in patients with coronary artery disease (CAD). However, it is not known whether beta-blockers have the same beneficial prognostic effect when residual ischemia persists on treatment. METHODS AND RESULTS: The prognostic impact of exercise single photon emission computed tomography (SPECT) ischemia was analyzed in 442 patients with chronic CAD, who were treated with beta blockers and who were referred to exercise thallium 201 SPECT, while they were receiving their daily-life medications. Ischemic and viable myocardium was documented on Tl-201 SPECT in 190 patients (43%), of whom only 23% had angina and only 26% had positive exercise testing results. During a follow-up of 3.8 +/- 1.7 years, 36 patients died and survival curves were progressively divergent between patients with and those without ischemic and viable myocardium: at 5 years, the respective survival rates were 81% +/- 4% and 94% +/- 2% (P =.004). By multivariate analysis, the best independent predictors of death were large extent of necrosis (>25% of left ventricle on Tl-201 SPECT, P <.001) and ischemic and viable myocardium (P =.001). CONCLUSIONS: In the CAD patients treated on a long term basis with beta-blockers, survival is strongly influenced by persistent exercise SPECT ischemia on treatment. Therefore exercise SPECT on treatment could be a useful tool for selecting those who might benefit from additional anti ischemic therapeutic interventions. PMID- 12900741 TI - Tl-201 myocardial SPECT in differentiation of ischemic from nonischemic dilated cardiomyopathy in patients with left ventricular dysfunction. AB - BACKGROUND: The differentiation between ischemic and nonischemic cardiomyopathy by noninvasive modalities is of clinical importance. Whether thallium 201 single photon emission computed tomography (SPECT) could accurately distinguish the two groups remains unclear. METHODS AND RESULTS: Twenty-nine patients with chronic heart failure (left ventricular ejection fraction < or =40%), including fourteen patients with ischemic cardiomyopathy and fifteen patients with nonischemic dilated cardiomyopathy, underwent Tl-201 SPECT. The stress protocols included treadmill exercise in 8 patients, dipyridamole in 6 patients, and dobutamine infusion in 15 patients. Myocardial SPECT was interpreted with the use of a 17 segment model and 0- to 4-point scale system. Patients with ischemic cardiomyopathy had higher summed stress defect scores (27.9 +/- 9.4 vs 20.6 +/- 8.9, P =.04), more fixed defect segments (5.9 +/- 2.9 vs 3.8 +/- 2.9, P =.05), and more moderate or severe perfusion defect segments on stress scan (7.2 +/- 2.0 vs 4.5 +/- 2.6, P =.004) than did those with nonischemic dilated cardiomyopathy. However, considerable overlap of the scan patterns between the two groups existed. Moderate or severe perfusion defects on stress scan in at least 7 segments were noted in 71% of patients (10/14) with ischemic cardiomyopathy, as compared with 20% of patients (3/15) with nonischemic cardiomyopathy (P =.016). CONCLUSIONS: Assessment of Tl-201 myocardial SPECT yields only modest value to distinguish nonischemic dilated cardiomyopathy from ischemic cardiomyopathy in patients with chronic heart failure. This technique cannot clearly differentiate individual patients. PMID- 12900742 TI - Accuracy of detection of myocardial viability and residual infarct vessel stenoses with rest Tl-201 and adenosine Tc-99m sestamibi imaging after coronary reperfusion in dogs with experimental acute myocardial infarction. AB - BACKGROUND: We sought to determine whether a dual-isotope imaging strategy (rest thallium 201/stress technetium 99m sestamibi) might be useful for assessing myocardial viability and residual ischemia in the infarct zone very early after reperfusion. METHODS AND RESULTS: Fifteen open-chest dogs had left anterior descending coronary artery occlusion for 60 minutes, followed by full reperfusion (group 1, n = 8) or reperfusion through a residual critical stenosis (group 2, n = 7). Tl-201 was injected at rest 45 minutes after reperfusion, and initial and 2 hour redistribution images were acquired. Tc-99m sestamibi was then injected during vasodilator stress, followed by imaging. Infarct size was similar in both groups (risk area, 21% +/- 4% vs 22% +/- 3%). Rest Tl-201 defect count ratios (left anterior descending coronary artery/left circumflex artery) were comparable (0.71 +/- 0.03 vs 0.74 +/- 0.02) and reflected infarct size. With vasodilation, Tc-99m sestamibi defect count ratio in group 1 (0.71 +/- 0.02) was comparable to rest Tl-201 and was significantly greater than in group 2 (0.62 +/- 0.02) with residual stenoses (P <.01). Although vasodilator Tc-99m sestamibi imaging unmasked the presence of residual stenoses, Tc-99m sestamibi uptake underestimated their functional severity (flow ratio, 0.38 +/- 0.03). CONCLUSIONS: Dual-isotope imaging very early after reperfusion may have limited utility for detecting residual stenoses in the infarct zone. Underestimation of the flow disparity by Tc-99m sestamibi may make the detection of stenoses more difficult, and impaired flow reserve after ischemic insult may complicate the detection of fully reperfused segments. PMID- 12900743 TI - Myocardial glucose uptake after dobutamine stress in chronic hibernating swine myocardium. AB - BACKGROUND: In patients with hibernating myocardium, regional uptake of the glucose analog 2-fluorine 18-fluoro-2-deoxy-d-glucose (FDG) is increased under resting conditions. It is unclear whether the degree of increased FDG uptake correlates with the degree of impaired blood flow response and whether chronic changes in the glucose transporters may play a role in the enhanced FDG uptake under fasted conditions. METHODS AND RESULTS: Twelve swine were instrumented with a constrictor on the left anterior descending (LAD) artery. Serial echocardiography and positron emission tomography studies were done to assess temporal changes in myocardial function, blood flow, and FDG uptake. One week after surgery (early study), wall thickening, blood flow, and postdobutamine FDG uptake in LAD and remote territories were similar. By approximately 6 weeks (late study), baseline wall thickening in the LAD region was lower than in remote regions (20% +/- 7% and 36% +/- 6%, P <.05), as was dobutamine-stimulated blood flow (0.92 +/- 0.16 mL. min(-1). g(-1) and 1.17 +/- 0.20 mL. min(-1). g(-1) in LAD and remote regions, respectively; P <.05). After the dobutamine infusion, FDG uptake in the LAD region during fasted conditions was higher than in remote regions (0.128 +/- 0.053 micromol. min(-1). g(-1) and 0.098 +/- 0.044 micromol. min(-1). g(-1), respectively; P <.05), and the increase was proportional to the impairment in dobutamine blood flow (r(2) = 0.62, P <.001). After the animals were killed, the LAD region showed a higher content of GLUT4 by immunoblots and a greater degree of translocation as estimated by immunohistochemistry. In 5 additional hibernating pigs studied under resting fasted conditions, FDG uptake and GLUT4 translocation were also higher in the LAD region, in the absence of dobutamine stimulation. CONCLUSIONS: In hibernating myocardium, regional FDG uptake under fasting conditions is higher than in remote regions, both at rest and after an infusion of dobutamine. The degree of poststress FDG uptake is proportional to the impaired stress-induced blood flow. Total GLUT4 content as well as membrane-bound protein is higher in the hibernating tissue, and these changes may facilitate the observed increase in FDG uptake. PMID- 12900744 TI - Noncardiac findings on dual-isotope myocardial perfusion SPECT. AB - BACKGROUND: This study examined the frequency of reporting noncardiac findings (NCFs), such as malignancies from inspection of raw projection images with dual isotope single photon emission computed tomography (SPECT) perfusion imaging, which could potentially be of greater clinical importance than myocardial perfusion imaging alone. Dual-isotope (ie, rest thallium 201 and stress technetium 99m sestamibi [MIBI] or Tc-99m tetrofosmin [TET]) SPECT myocardial perfusion imaging combines multipotential tracers for noncardiac purposes (Tl-201 for renal or splenic imaging, inflammation, or lymphoma and MIBI or TET for hepatobiliary imaging and detecting increased mitochondrial number or activity in neoplastic processes). These images are optimally interpreted with cinematic inspection of the raw projection data, but this may not be practiced uniformly in every laboratory. METHODS AND RESULTS: We reviewed 12,526 computer-generated text reports of dual-isotope perfusion SPECT studies from a 6-year period for NCFs. NCFs were categorized by organ and by probability of malignancy: high (eg, focal breast or lung uptake of MIBI or TET), intermediate (eg, lymph node uptake or thyroid abnormalities), or low (eg, filling defects in liver, kidney, spleen, or gall bladder; ascites; or pleural effusions). Confirmatory imaging studies or clinical confirmation for each NCF was sought. There were a total of 207 NCFs identified in 180 reports (1.7% of reports, ranging from 0% to 2.8% of reports of individual interpreters). Of these, 107 NCFs were unsuspected before SPECT; 24% were considered at high probability for malignancy, and 24% were considered intermediate in likelihood of malignancy. Follow-up data were available for 178 NCFs, confirming 88% of these findings, including 82% of breast foci, 62% of lung foci, 86% of hepatobiliary/spleen abnormalities, and 94% of renal abnormalities. The probability of malignancy was highest (82%) in breast or lung foci in which uptake of both Tl-201 and the Tc-99m-labeled agent was present. CONCLUSIONS: In patients referred for evaluation of myocardial perfusion, NCFs are unusual and require systematic and careful inspection of projection images for their detection. With Tl-201, TET, MIBI, or dual-isotope imaging, detecting and reporting NCFs may occasionally result in life-saving early cancer identification. PMID- 12900745 TI - Stem cells and cardiovascular disease. AB - Several recent discoveries have shifted the paradigm that there is no potential for myocardial regeneration and have fueled enthusiasm for a new frontier in the treatment of cardiovascular disease-stem cells. Fundamental to this emerging field is the cumulative evidence that adult bone marrow stem cells can differentiate into a wide variety of cell types, including cardiac myocytes and endothelial cells. This phenomenon has been termed stem cell plasticity and is the basis for the explosive recent interest in stem cell-based therapies. Directed to cardiovascular disease, stem cell therapy holds the promise of replacing lost heart muscle and enhancing cardiovascular revascularization. Early evidence of the feasibility of stem cell therapy for cardiovascular disease came from a series of animal experiments demonstrating that adult stem cells could become cardiac muscle cells (myogenesis) and participate in the formation of new blood vessels (angiogenesis and vasculogenesis) in the heart after myocardial infarction. These findings have been rapidly translated to ongoing human trials, but many questions remain. This review focuses on the use of adult bone marrow derived stem cells for the treatment of ischemic cardiovascular disease and will contrast how far we have come in a short time with how far we still need to go before stem cell therapy becomes routine in cardiovascular medicine. PMID- 12900746 TI - Pulmonary hypertension: work in progress. AB - Pulmonary arterial hypertension is a rare disorder defined by mean pulmonary artery pressures that exceed 25 mm Hg at rest or are greater than 30 mm Hg with exercise. The mortality rate is high for untreated patients, mainly as a result of progressive right heart dysfunction. Pulmonary arterial hypertension has been historically classified as primary pulmonary hypertension or pulmonary hypertension resulting from an underlying disease process. Ongoing research in the nuclear medicine field holds great promise for understanding the pathophysiologic pathways for this condition, as well as the monitoring of the continually evolving therapeutic options. PMID- 12900748 TI - Effects of therapy with beta-blocker agents on myocardial perfusion and outcome. PMID- 12900747 TI - Scintigraphic blood pool and phase image analysis: the optimal tool for the evaluation of resynchronization therapy. PMID- 12900749 TI - Antianginal medications and diagnostic accuracy of myocardial perfusion imaging. PMID- 12900750 TI - Increased lung uptake on exercise Tl-201 SPECT may reflect more than extent of myocardial ischemia. PMID- 12900751 TI - Recurrent myocardial infarction with near-normal coronary angiogram and myocardial ischemia detected by Tc-99m SPECT and magnetic resonance perfusion imaging. PMID- 12900753 TI - UK cash for gene therapy: government pledges support. PMID- 12900755 TI - T-cell activation by recombinant immunoreceptors: impact of the intracellular signalling domain on the stability of receptor expression and antigen-specific activation of grafted T cells. AB - Recombinant immunoreceptors are modularily composed of extracellular antigen binding and intracellular signalling domains that are preferentially derived from CD3zeta or Fc epsilon RIgamma. The impact of the signalling domain on the stability of immunoreceptor expression and function is not completely understood. To address this issue, we generated and expressed a panel of recombinant zeta- and gamma-chain immunoreceptors, respectively, in human peripheral blood T cells. The expression level of zeta-chain immunoreceptors in human T cells is significantly lower than those of the homologous gamma-chain receptors. Low zeta chain receptor expression in peripheral T cells is because of the intracellular signalling domain and independent of the Fc epsilon RIgamma or CD3zeta transmembrane region. Expression of both receptors decreases upon prolonged cultivation. Shortly after receptor engraftment, target cell lysis and induction of IFN-gamma secretion are mediated with similar efficiency by zeta- and gamma chain immunoreceptors. Upon prolonged propagation, however, T-cell activation mediated by zeta-chain immunoreceptors is more efficient than by gamma-chain receptors, indicating that the initial high expression level of gamma-chain immunoreceptors compensates its lower activation capacity. Consequently, gamma chain immunoreceptors exhibit a higher threshold value for specific activation and are more pronouncedly inhibited by soluble ligand antigen compared to the homologous zeta-chain receptor. These findings have substantial consequences for the design of recombinant immunoreceptors for use in adoptive immunotherapy. PMID- 12900756 TI - DNA delivery from an intravascular stent with a denatured collagen-polylactic polyglycolic acid-controlled release coating: mechanisms of enhanced transfection. AB - We previously demonstrated that DNA-polylactic-polyglycolic acid (PLGA)-coated stents can deliver genes to the arterial wall with reporter expression involving 1% of neointimal cells. The present study investigated a novel formulation utilizing denatured collagen in DNA-stent coatings; denatured collagen was hypothesized to enhance gene transfer due to adhesion molecule interactions and actin-related mechanisms. Arterial smooth muscle cells (SMCs) cultivated on denatured collagen had significantly greater plasmid DNA (beta-galactosidase) transfection than SMC grown on native collagen (18.3+/-1.2 vs 1.0+/-0.1%, P<0.001). The denatured-collagen effect was completely blocked with anti alpha(v)beta(3) integrin antibody. SMCs cultivated on native collagen supplemented with tenascin-C (TN-C), a protein recognized by alpha(v)beta(3) integrins, showed a 33-fold increase in transfection compared to control (P<0.001); this effect was also blocked with anti-alpha(v)beta(3) antibody. We observed that cells grown on denatured collagen had marked F-actin-enriched stress fibers and intense perinuclear G actin, compared to those grown on native collagen, which demonstrated F-actin-enriched focal adhesions without perinuclear G-actin localization. Cytochalasin-D, an F actin depolymerizing agent, caused significantly increased SMC transfection in cells cultivated on native collagen compared to control cells (18.0+/-1.8 vs 3.02+/-0.9%, P<0.001) further supporting the view that actin-related cytoskeletal changes influence transfection. A denatured-collagen-PLGA composite vascular stent coating similarly resulted in increased plasmid DNA green fluorescent protein (GFP) expression compared to controls (P<0.001) in SMC cultures; the increased transfection was blocked by anti-alpha(v)beta(3) antibody. Pig coronary studies comparing denatured-collagen PLGA-coated stents containing plasmid DNA (encoding GFP) to coated stents without DNA demonstrated 10.8% of neointimal cells transfected; this level of expression was almost an order of magnitude greater than previously reported with a DNA delivery stent. It is concluded that denatured collagen incorporated into plasmid DNA-stent coating formulations may increase the level of gene expression in vitro and in vivo because of integrin-related mechanisms and associated changes in the arterial smooth muscle cell actin cytoskeleton. PMID- 12900757 TI - Mapping alveolar binding sites in vivo using phage peptide libraries. AB - Targeting lung tissue is nonselective due in part to the lack of specific cell surface receptors identified on target lung cells. We used in vivo phage display to identify a panel of peptides that can bind selectively to lung epithelial cells with less binding to nonepithelial cells. By direct intratracheal instillation of phage libraries into the lung, we isolated and identified 143 individual phage clones. Three phage clones revealed enhanced binding to the lung in vitro and in vivo. These three identified peptides were synthesized and demonstrated selective binding to epithelial cells in lung tissue versus the control peptide. Further, the peptides specifically bound to freshly isolated type II alveolar epithelial cells compared with Hep2 cells. The results suggest that the airway phage display approach could be exploited for analyzing the molecular diversity in the lower respiratory tract. PMID- 12900758 TI - Viral oncoapoptosis of human tumor cells. AB - Many cancer cells refractory to radiation treatment and chemotherapy proliferate because of loss of intrinsic programmed cell death (apoptosis) regulation. Consequently, the resolution of these cancers are many times outside the management capabilities of conventional therapeutics. We now report that replication-defective delta27 herpes simplex virus (rd delta27) triggers apoptosis in three representative transformed human cell lines. Susceptibility to virus-induced cell death is dependent on the abundance and distribution of modified p53 protein in the tumor cells indicating specific targeting of the treatment. Primary human and mouse fibroblast cells that produce modified p53 are resistant to rd delta27 killing but not to apoptosis induced by nonviral environmental factors. These results suggest that induction of apoptosis by nonreplicating virus is a feasible genetic therapy approach for killing human cancer cells. Our findings may have important implications in designing novel virus-based anticancer strategies in appropriate animal model systems. PMID- 12900759 TI - Effective transduction and stable transgene expression in human blood cells by a third-generation lentiviral vector. AB - Difficulty in gene transduction of human blood cells, including hematopoietic stem cells, has hampered the development of gene therapy applications for hematological disorders, encouraging the development and use of new gene delivery systems. In this study, we used a third-generation self-inactivating (SIN) lentiviral vector system based on human immunodeficiency virus type 1 (HIV-1) to improve transduction efficiency and prevent vector-related toxicity. The transduction efficiency of the HIV-1-based vector was compared directly with the Moloney murine leukemia virus (MLV) SIN vector in human leukemia cell lines. Initial transduction efficiencies were almost 100% for the HIV and less than 50% for the MLV vectors. Similar results were observed in 11 types of primary cells obtained from leukemia or myeloma patients. Transgene expression persisted for 8 weeks in cells transduced with the HIV vector, but declined with the MLV vector. In addition, resting peripheral blood lymphocytes and CD34(+) hematopoietic cells were transduced successfully with the HIV vector, but not with the MLV vector. Finally, we confirmed vector gene integration in almost all colony-forming cells transduced with the HIV vector, but not with the MLV vector. In conclusion, this lentiviral vector is an excellent gene transduction system for human blood cells because of its high gene transduction and host chromosome integration efficiency. PMID- 12900760 TI - A strict-late viral promoter is a strong tumor-specific promoter in the context of an oncolytic herpes simplex virus. AB - Confinement of transgene expression to target cells is highly desirable in gene therapy. Current strategies of transcriptional targeting to tumors usually rely on tissue-specific promoters to control gene expression. However, such promoters generally have much lower activity than the constitutive viral promoters. We have explored an alternative approach, using a strict-late viral promoter (UL38p) in the context of an oncolytic herpes simplex virus (HSV) for tumor-selective gene expression. As with many DNA viruses, the genomic transcription of HSV is a tightly regulated molecular cascade in which early and late phases of gene expression are separated by viral DNA replication. In particular, some of the late transcripts are categorized as strict-late, whose expression depends rigorously on the initiation of viral DNA replication. Our in vitro and in vivo characterization showed that in normal nondividing cells, where the oncolytic HSV has limited ability to replicate, the UL38p has minimal activity. However, in tumor or cycling cells where the virus can fully replicate, transgene expression from UL38p was almost as high as from the cytomegalovirus immediate-early promoter. These results suggest that delivery of therapeutic genes driven by UL38p through an oncolytic HSV may be an effective approach to gene therapy for malignant diseases. PMID- 12900761 TI - Effect of a DNA nuclear targeting sequence on gene transfer and expression of plasmids in the intact vasculature. AB - Although the use of nonviral vectors for gene therapy offers distinct advantages including the lack of significant inflammatory and immune responses, the levels of expression in vivo remain much lower than those obtained with their viral counterparts. One reason for such low expression is that unlike many viruses, plasmids have not evolved mechanisms to target to the nucleus of the nondividing cell. In the absence of mitosis, plasmids are imported into the nucleus in a sequence-specific manner, and we have shown in cultured cells by transfection and microinjection experiments that the SV40 enhancer mediates plasmid nuclear import in all cell types tested (Dean et al., 1999, Exp Cell Res 253: 713-722). To test the effect of this import sequence on gene transfer in the intact animal, we have recently developed an electroporation method for DNA delivery to the intact mesenteric vasculature of the rat. Plasmids expressing luciferase or GFP from the CMV immediate-early promoter/enhancer and either containing or lacking the SV40 enhancer downstream of the reporter gene were transferred to the vasculature by electroporation. When transfected into actively dividing populations of smooth muscle or epithelial cells, the plasmids gave similar levels of expression. By contrast, the presence of the SV40 sequence greatly enhanced gene expression of both reporters in the target tissue. At 2 days post-transfer, plasmids with the SV40 sequence gave 10-fold higher levels of luciferase expression, and at 3 days the difference was over 40-fold. The presence of the SV40 sequence did not simply increase the rate of nuclear import and expression, since expression from the SV40-lacking plasmid did not increase beyond that seen at day 2, the time of maximum expression for either plasmid. In situ hybridization experiments confirmed that the increased gene transfer and expression was indeed due to increased nuclear localization of the delivered SV40 sequence-containing plasmid. Based on these findings, the ability to target DNA to the nucleus can increase gene transfer in vivo and inclusion of the SV40 sequence into plasmids will enhance nonviral gene delivery. PMID- 12900762 TI - Transgene delivery of plasmid DNA to smooth muscle cells and macrophages from a biostable polymer-coated stent. AB - Metallic stents coated with a polyurethane emulsion containing plasmid DNA were implanted in rabbit iliac arteries to evaluate transgene delivery and expression in the vessel wall. The expression of the plasmid-encoded marker genes, beta galactosidase, luciferase and green fluorescence protein (GFP), were evaluated at 7 days after implantation. In all cases, plasmid transfer was confined to the vessel wall at the site of stent implantation, plasmid DNA was not observed in vessel segments immediately proximal or distal to the stent and dissemination of plasmid DNA to lung, liver or spleen was not observed. Expression of transgenes occurred only in vessel segments in contact with the stent and analysis of the GFP expression pattern revealed a high frequency of marker protein-positive cells occurring at or near the luminal surface. The extent of transgene expression was dependent upon the quantity of DNA loaded onto the stent and no signal was detected in vessel segments that received polymer-coated stents lacking plasmid DNA. Of significance, colocalization studies identified transgene expression not only in vascular smooth muscle cells but also in macrophages. Hence, polymer coated stents provide a new capability for transgene delivery to immune cells that are believed to contribute to the development of in-stent restenosis. PMID- 12900763 TI - Gene therapy to target dendritic cells from blood to lymph nodes. AB - Peripheral lymph nodes (PLN) are strategic microenvironments where antigen presenting dendritic cells (DC), loaded with environmental antigens, and naive lymphocytes meet to initiate immune responses. The unique capacity of DC to induce primary immune responses has led to their use in clinical medicine; however, delivering DC to lymph nodes is problematic. Intravenously injected DC cannot access to PLN, while DC injected into tissue migrate inefficiently through lymphatics to PLN. We achieved DC targeting to T-cell areas of PLN by endowing DC with a novel receptor for peripheral node addressin (PNAd), an adhesion molecule present on the lymph node venular endothelium. This novel receptor is a chimeric E/L-selectin (ELS) that, we have previously shown, binds to PNAd. DC were genetically modified by retroviral transduction to express ELS. ELS expression was targeted to tips of microvilli, and mediated rolling of DC on PNAd both in vivo and in vitro. Such genetically engineered DC could extravasate directly from blood through the lymph node endothelium as opposed to nontransduced DC. This study provides evidence that the trafficking of DC can be modified using gene transfer technologies. More efficient delivery of DC to PLN should assist the development of improved vaccination strategies. PMID- 12900764 TI - Attenuation of ganglioside GM1 accumulation in the brain of GM1 gangliosidosis mice by neonatal intravenous gene transfer. AB - A single intravenous injection with 4 x 10(7) PFU of recombinant adenovirus encoding mouse beta-galactosidase cDNA to newborn mice provided widespread increases of beta-galactosidase activity, and attenuated the development of the disease including the brain at least for 60 days. The beta-galactosidase activity showed 2-4 times as high a normal activity in the liver and lung, and 50 times in the heart. In the brain, while the activity was only 10-20% of normal, the efficacy of the treatment was distinct. At the 30th day after the injection, significant attenuation of ganglioside GM1 accumulation in the cerebrum was shown in three out of seven mice. At the 60th day after the injection, the amount of ganglioside GM1 was above the normal range in all treated mice, which was speculated to be the result of reaccumulation. However, the values were still definitely lower in most of the treated mice than those in untreated mice. In the histopathological study, X-gal-positive cells, which showed the expression of exogenous beta-galactosidase gene, were observed in the brain. It is noteworthy that neonatal administration via blood vessels provided access to the central nervous system because of the incompletely formed blood-brain barrier. PMID- 12900765 TI - Herpes simplex virus type-1 infection upregulates cellular promoters and telomerase activity in both tumor and nontumor human cells. AB - Targeted gene expression through viral vectors has been a promising approach for gene therapy. However, the effects of viral gene products expressed from virus vectors on the expression of the host gene are not well known. In the present study, we examined the activities of cellular promoters, including the promoter for genes of human telomerase reverse transcriptase (hTERT), tyrosinase and probasin, in both tumor and normal cells after infection with herpes simplex virus type 1 (HSV-1) vectors. Our results showed that infection with replication defective HSV-1 vectors significantly upregulated the activity of all three cellular promoters in a nonsequence specific fashion in all cell types tested. Furthermore, viral infection upregulated activities of the hTERT promoter and endogenous telomerase in nontumoral cells. Additional experiments suggested that the viral immediate-early gene product, infected cell protein 0, might be responsible for the deregulation of cellular promoter activity and activation of telomerase. Our study alerts to the potential risk of oncogenesis through deregulation of host gene expression, such as the telomerase by viral vectors in normal cells. PMID- 12900766 TI - Simian lentiviral vector-mediated retinal gene transfer of pigment epithelium derived factor protects retinal degeneration and electrical defect in Royal College of Surgeons rats. AB - Retinitis pigmentosa (RP) is a heterogenous group of inherited retinal diseases resulting in adult blindness caused by mutations of various genes. Although it is difficult to cure the blindness that results from these diseases, delaying the disease progression may be of great benefit, since the majority of RP diseases are seen in middle age or later. To test a gene therapy strategy for RP using a neurotrophic factor gene, we assessed the effect of simian lentivirus (SIV) mediated subretinal gene transfer of pigment epithelium-derived factor (PEDF), a potent neurotrophic factor, during the disease progression in Royal College of Surgeons (RCS) rats, a well-accepted animal model of RP. Regional gene transfer via SIV into the peripheral subretinal space at the nasal hemisphere was performed in all animals to monitor site-specific transgene expression as well as the therapeutic effect in each retina. Gene transfer of lacZ and PEDF was observed in the regional pigment epithelium corresponding to the regional gene transfer. Histologically, PEDF gene transfer significantly protected the loss of photoreceptor cells (PCs) corresponding to the regions of the gene transfer, compared to those of control groups during the course of the experiment. The antiapoptotic effect of PEDF on PCs is likely to be a related mechanism, because a significant reduction of terminal dUTP-nicked end labeling-positive PC numbers was found in PEDF-treated eyes compared to those of the control group (P<0.05). PEDF-treated eyes also retained a significant sensitivity to light flash during the experimental course. These findings clearly show that neuroprotective gene therapy using PEDF can protect retinal degeneration and functional defects in individuals with RP. PMID- 12900767 TI - Generating MHC Class II+/Ii- phenotype after adenoviral delivery of both an expressible gene for MHC Class II inducer and an antisense Ii-RNA construct in tumor cells. AB - Tumor cells engineered by gene transduction to be MHC Class II+/Ii- are novel APCs capable of presenting endogenous tumor antigen epitopes to activate T helper cells. The MHC Class II+/Ii- tumor cell phenotype is created by transfecting genes for either CIITA or IFN-gamma, and inhibiting induced Ii mRNA by an Ii reverse gene construct (Ii-RGC). Adenoviral vectors are preferred for the delivery of such genes because of high infection efficiency and ubiquity of the adenoviral receptor on many cell types and tumors. Here we show that at 5 MOI (multiplicity of infection), recombinant adenoviruses with CIITA or IFN-gamma genes converted virtually all MC-38 colon adenocarcinoma cells and Renca renal carcinoma cells in culture to MHC Class II+/Ii+ cells. A single recombinant adenovirus with both genes for IFN-gamma and Ii-RGC (rAV/IFN-gamma/Ii-RGC) efficiently induced the MHC Class II+/Ii- phenotype. Injection of tumor nodules with rAV/Ii-RGC and rAV/CIITA/IFN-gamma combined with a suboptimal dose of rAV/IL 2 induced a potent antitumor immune response. The methods are adaptable for producing enhanced genetic vaccines, attenuated virus vaccines (eg, vaccinia), and ex vivo cell-based vaccines (dendritic and tumor cells). PMID- 12900768 TI - Directed apoptosis in Cox-2-overexpressing cancer cells through expression targeted gene delivery. AB - The principle of promoter-targeted gene delivery was used to direct the expression of reporter genes and inducible caspases to Cox-2-overexpressing cancer cells. The polycation poly(ethylenimine) was used in unmodified form to nonvirally deliver genes into cells, and targeting was achieved at the transcriptional level. Results demonstrated that reporter expression was reduced by an average of 89.8% in normal cells and cell lines not overexpressing Cox-2 when the strong cytomegalovirus promoter was replaced with the human Cox-2 promoter in delivered plasmids. Cocultures of normal and Cox-2-overexpressing cancer cells showed less than 0.5% reporter expression in normal fibroblast cells but over 35% reporter expression in PC3 prostate cancer cells. This targeting method was then used to direct the expression of inducible forms of caspases 3 and 9 to Cox-2-overexpressing cancer cells of the bladder and prostate. Following activation of the resulting caspase pro-forms, cells underwent apoptosis as evidenced by DNA fragmentation and cytoskeletal degradation. This result was also observed in cells resistant to apoptosis in terms of TNF-alpha initiation. Such directed apoptosis could eventually serve as a treatment for an entire class of Cox-2-overexpressing carcinomas. PMID- 12900769 TI - Recombinant AAV serotype 1 transduction efficiency and tropism in the murine brain. AB - Recombinant adeno-associated virus serotype 2 (rAAV2) vectors have shown promise as therapeutic agents for neurologic disorders. However, intracerebral administration of this vector leads to preferential transduction of neurons and a restricted region of transgene expression. The recently developed rAAV vectors based upon nonserotype 2 viruses have the potential to overcome these limitations. Therefore, we directly compared a rAAV type 1 to a type 2 vector in the murine brain. The vectors were engineered to carry identical genomes (AAV2 terminal repeat elements flanking an enhanced green fluorescent protein expression cassette) and were administered by stereotaxic-guided intracerebral injection. We found that the rAAV1 vector (rAAV1-GFP) had a 13- to 35-fold greater transduction efficiency than that of the rAAV2 vector (rAAV2-GFP). Also, rAAV1-transduced cells were observed at a greater distance from the injection site than rAAV2-transduced cells. Neurons were the predominant cell type transduced by both vector types. However, in contrast to rAAV2-GFP, rAAV1-GFP was capable of transducing glial and ependymal cells. Thus, rAAV1-based vectors have biologic properties within the brain distinct from that of rAAV2. These differences might be capitalized upon to develop novel gene transfer strategies for neurologic disorders. PMID- 12900770 TI - N-acetyl-L-cysteine does not affect the pharmacokinetics or myelosuppressive effect of busulfan during conditioning prior to allogeneic stem cell transplantation. AB - Busulfan is currently used as a main component in the conditioning regimen prior to allogeneic stem cell transplantation (SCT). Several studies have shown a correlation between exposure to busulfan and transplantation-related liver toxicity, such as venoocclusive disease (VOD) in patients undergoing SCT. Busulfan is metabolized mainly through glutathione (GSH). During high-dose therapy, busulfan may deplete hepatocellular levels of GSH. As part of the conditioning therapy, busulfan is usually followed by high doses of cyclophosphamide. The activation of cyclophosphamide yields a cytotoxic metabolite, 4-hydroxy cyclophosphamide, which is highly reactive and detoxified through GSH. According to recent studies using cell lines and animal models N acetyl-L-cysteine (NAC), a GSH precursor, does not hamper the myeloablative effect of busulfan during conditioning. In the present study, we administered NAC during conditioning to 10 patients at risk of VOD due to pretransplant liver disorders or elevated liver enzymes. No side effects related to the NAC infusions were observed and busulfan concentrations were not affected. All patients became pancytopenic and engrafted with 100% donor cells. None of the patients developed VOD or liver failure. Increased liver enzymes during conditioning decreased or normalized in all patients. We suggest that NAC therapy is safe and does not impair the myeloablative effect of busulfan during conditioning prior to SCT. PMID- 12900771 TI - ATG as part of the conditioning regimen reduces transplant-related mortality (TRM) and improves overall survival after unrelated stem cell transplantation in patients with chronic myelogenous leukemia (CML). AB - Matched unrelated donor transplants have an increased risk of severe graft-versus host disease and transplant-related mortality (TRM). ATG has been introduced to decrease GvHD and to facilitate engraftment. We conducted a retrospective analysis of 333 patients with chronic myelogenous leukemia, who were treated with Fresenius ATG (n=145, average=90 mg/kg bw, range 40-90 mg/kg bw) or standard immunosuppression without ATG (n=188). Both groups were comparable regarding distribution of age, sex, HLA-matched vs mismatched donors. ATG Fresenius led to a faster leukocyte engraftment, decreased the incidence of acute GvHD and TRM (P=0.01 and P=0.03) and led to a significant better overall survival (70 vs 57%, P=0.03). We concluded that a prospective randomized study is needed to evaluate the definite role of ATG in hemopoietic stem cell transplantation. PMID- 12900772 TI - Induction of oral tolerance in bone marrow transplantation recipients suppresses graft-versus-host disease in a semiallogeneic mouse model. AB - Graft-versus-host disease (GVHD) is the major obstacle for successful allogeneic stem cell transplantation (SCT). Morbidity and mortality are high, and novel therapeutic strategies are required. Current therapy, which is based mainly on immunosuppression, is associated with a high degree of complications. Immune hyporesponsiveness induced by oral antigen administration has recently been shown to prevent the development of chronic GVHD (cGVHD) in a murine model. The aim of the present study was to evaluate whether it is possible to induce tolerance and to alleviate GVHD in a semiallogeneic transplantation model in mice. GVHD was generated by infusing 2 x 10(7) splenocytes from C57BL/6 donor mice into (C57BL/6 x Balb/c)F1 recipient mice, which received 7 Gy (60)Co total body irradiation (TBI) prior to transplantation. Oral tolerance was induced by feeding recipient F1 mice with five oral doses of proteins, 50 micro g/mouse, extracted from C57BL/6 splenocytes on alternate days following transplantation. In vitro mixed lymphocyte reaction (MLR) from tolerized and nontolerized mice was performed. Recipient mice were followed for chimerism, and for clinical and histological parameters of GVHD. Induction of tolerance was documented by a significant reduction in MLR response of tolerated vs nontolerated splenocytes. A significant alleviation of the clinical and pathological manifestation of GVHD was observed in the liver, small bowel, and skin. Tolerance induction did not jeopardize engraftment. These results may constitute a step towards reducing the frequency of GVHD via manipulation of the immune system. PMID- 12900773 TI - Tissue-restricted T cell alloresponses across HLA barriers: selection and identification of leukemia-restricted CTL in HLA-mismatched stimulator-responder pairs. AB - Exploiting the graft-versus-leukemia (GVL) effect in mismatched transplants requires its separation from graft-versus-host disease (GVHD). We generated leukemia-specific cytotoxic T lymphocytes (CTL) in three haplotype-mismatched, two class I-mismatched and two single HLA-A locus-matched stimulator-responder pairs. Six patients with chronic myelogenous leukemia and one patient with acute myeloid leukemia transformed from MDS were studied. CTL generated after 10 days stimulation with unselected leukemic peripheral blood mononuclear cells inhibited leukemic CFU-GM colony growth (>85% at 10:1 effector:target ratio) with no third party colony inhibition. In five pairs, responders were cultured separately with leukemia cells, PHA-B or LCL from the stimulator. After 2-4 restimulations, the T cell repertoire was examined by flow analysis using Vbeta-specific antibodies. Test cultures (but not controls) showed preferential expansion of 1-4 Vbeta families either common to two or more stimulators or unique to a particular stimulator. Notably, we elicited leukemia-specific TCR Vbeta expansions on four out of five occasions. In two pairs, responder cells selected for the appropriate leukemia-specific Vbeta family were shown to have leukemia-specific cytotoxicity. These leukemia-restricted T-cells were CD8+ or CD4+ and CD25+ or CD57+. The results support the development of strategies to selectively deplete GVHD and conserve GVL reactivity in mismatched transplants. PMID- 12900774 TI - A prospective comparison of immune reconstitution in pediatric recipients of positively selected CD34+ peripheral blood stem cells from unrelated donors vs recipients of unmanipulated bone marrow from related donors. AB - Positively selected CD34(+) hematopoietic stem cells from unrelated donors (UD HSCT) have been successfully transplanted, but little is known about immune reconstitution in this setting. Here we report a prospective comparison of immune reconstitution in recipients of UD-HSCT and of unmanipulated bone marrow from matched sibling donors (MSD-BMT). T-cell reconstitution occurred more than 100 days later in the UD-HSCT than in the MSD-BMT group. The first T cells after UD HSCT were almost exclusively CD45RO(+) HLA-DR(+), whereas early-emerging T cells after MSD-BMT more frequently expressed CD62L, CD28, and CD25. In both groups, numbers of CD45RA(+) naive T cells increased after 180 days. After UD-HSCT, the T cell-receptor (TCR)-repertoire was severely skewed and showed significantly reduced diversity during the first year, but only minor abnormalities were seen after MSD-BMT. TCR-diversity increased simultaneously with the number of naive T cells. In both groups, we observed transient expansions of gammadelta T cells. B cells were reconstituted more rapidly in UD-HSCT than in MSD-BMT recipients, whereas the rapidity of NK-cell reconstitution was similar in the two groups. In summary, T-cell reconstitution was slower after UD-HSCT than after MSD-BMT because of the delayed recovery of early memory-type T cells with reduced TCR diversity, whereas naive T-, NK-, and B cells were reconstituted similarly in the two groups. PMID- 12900775 TI - Serum stem cell growth factor for monitoring hematopoietic recovery following stem cell transplantation. AB - Stem cell growth factor (SCGF) is a novel cytokine for primitive hematopoietic progenitor cells. Although it has burst-promoting activity and granulocyte/macrophage colony-promoting activity in vitro, its significance in hematopoiesis in vivo has not been elucidated. In this study, we have established enzyme-linked immunosorbent assay (ELISA) to quantify human SCGF and measured serum cytokines in normal volunteers and 27 patients undergoing stem cell transplantation (SCT), including six autologous and 21 allogeneic transplants. SCGF levels gradually increased after SCT regardless of graft-versus-host disease or type of transplant. The maximum level of SCGF was observed during the rapid granulocyte recovery phase in patients subjected to an autologous transplantation, and during the granulocyte stabilization phase in allogeneic patients. SCGF levels in PBSCT patients began to rise earlier than in BMT patients. Two patients with no increment of SCGF after SCT showed delayed engraftment. The source of SCGF was further analyzed by RT-PCR and we found that SCGF was highly expressed in bone marrow (BM) CD34(+) and CD34(-)CD33(+) cells, but not in BM CD34(-)CD33(-) cells, BM stromal cells and peripheral blood cells. The cell population expressing SCGF in BM possess the colony-forming cell activity. Therefore, serum SCGF can be an indicator of hematopoietic recovery following SCT. PMID- 12900776 TI - Pegfilgrastim, a sustained-duration form of filgrastim, significantly improves neutrophil recovery after autologous marrow transplantation in rhesus macaques. AB - Daily administration of filgrastim decreases the duration of severe neutropenia in the clinical setting. A sustained-duration form of filgrastim, pegfilgrastim, significantly reduces scheduling protocols to a single injection per chemotherapy cycle while maintaining therapeutic efficiency. We examined the ability of a single injection of pegfilgrastim to significantly improve neutrophil recovery following autologous bone marrow transplantation (AuBMT) in rhesus macaques. On day 1, postmyeloablation (920 cGy x-irradiation) and AuBMT, animals received either 0.1% autologous serum for 18 consecutive days (n=13), or single doses of pegfilgrastim via the subcutaneous (s.c.) or intravenous (i.v.) route (300 or 100 micro g/kg), or a single dose of filgrastim at 300 micro g/kg via the s.c. or i.v. route, or filgrastim at 10 micro g/kg via the s.c. route (n=4) on a daily basis (range=days 12-17). Pharmacokinetic parameters and neutrophil recovery were assessed. A single dose of pegfilgrastim via the i.v. or s.c. route was as effective as daily filgrastim administration, resulting in significant improvement of neutrophil recovery after myeloablation and ABuMT. Effective pegfilgrastim plasma concentrations were maintained in neutropenic animals until after the onset of hematopoietic recovery. Enhanced pharmacokinetics in AuBMT cohorts are consistent with self-regulating, neutrophil-mediated clearance. PMID- 12900777 TI - Acute renal failure requiring dialysis after allogeneic blood and marrow transplantation identifies very poor prognosis patients. AB - We examined the incidence, risk factors and associated mortality of acute renal failure requiring dialysis (Renal Bearman Grade [BG] 3) in a 3-year cohort of 97 consecutive allogeneic blood and marrow transplantation (alloBMT) patients. In all, 20 (21%) developed Renal BG3 (all died by day +132) and 77 (79%) developed renal insufficiency (Renal BG1-2). Renal BG3 was a contributing or primary cause of death in 18 (90%) patients who continued to require dialysis at time of death. The two Renal BG3 patients whose deaths were not related to renal failure died on day +103 of hemorrhage and day +132 of underlying disease. By univariate analysis, age, unrelated donor, veno-occlusive disease (VOD) and grade III-IV acute graft-versus-host disease with hepatic involvement were significantly associated with Renal BG3. The multivariate model of time to Renal BG3 determined only a prior diagnosis of severe acute GVHD (RR=4.1, 95% CI 1.6-10.3, P=0.003) and VOD (RR=9.1, 95% CI 3.5-23.7, P<0.001) as significant independent predictors. Renal BG3 is generally considered a conditioning regimen-related toxicity. This study demonstrates that Renal BG3 is most commonly a complication of hepatic co morbidities after allogeneic blood and marrow transplantation and identifies patients with a very poor prognosis. PMID- 12900778 TI - Changes of serum growth factors (IGF-I,-II and IGFBP-2,-3) prior to and after stem cell transplantation in children with acute leukemia. AB - SUMMARY: Insulin-like growth factors (IGFs) and IGF-binding proteins (IGFBPs) may play an important role in tumor proliferation. This study aimed to investigate the IGF system in children with acute leukemia prior to and after hematological stem cell transplantation (HSCT). In 51 patients (AML n=27; ALL n=24; mean age 11.2+/-4.8 years), serum parameters (IGF-I,-II, IGFBP-2,-3) were investigated up to 18 months after HSCT by RIA. Patients with AML showed a significant increase of IGFBP-2 up to 100 days after HSCT (mean +/-s.d. prior to HSCT: 3.2+/-3.6 SDS vs 100 days after HSCT: 5.3 degrees +/-3.4 SDS, P=0.005). Furthermore, IGF-I and IGFBP-3 were significantly decreased (IGF-I: -0.3+/-1.5 vs -0.7 +/-1.2 SDS, P=0.001; IGFBP-3: -0.3+/-1.1 vs -1.0+/-1.1 SDS, P=0.02). Children with AML showed significantly higher IGFBP-2 (P=0.04) and significantly lower IGF-I (P=0.03) and IGFBP-3 (P=0.05) levels than children with ALL at day 100 after HSCT. We conclude that children with acute leukemia show important changes in the IGF system after HSCT. In particular, IGFBP-2 was significantly elevated at day 100 after HSCT. Increased IGFBP-2 and decreased IGF-I and IGFBP-3 may be associated with the increased proliferation rate of transplanted bone marrow. PMID- 12900779 TI - Clinical features and treatment of hematopoietic stem cell transplantation associated gastric antral vascular ectasia. AB - Gastric antral vascular ectasia (GAVE) may occur after hematopoietic stem cell transplantation (HSCT) and cause severe and prolonged gastric bleeding. The underlying pathology of transplant-associated GAVE (HSCT-GAVE) is poorly understood and an effective therapeutic strategy has not been established yet. We retrospectively reviewed the medical records of 230 consecutive allogeneic transplant recipients in our institution between January 1997 and June 2002. We identified five patients who developed HSCT-GAVE (2.2%). Four patients had bleeding from HSCT-GAVE and one patient had HSCT-GAVE discovered incidentally. The clinical features of these patients were similar in that they all received conditioning treatment with busulfan and had history of thrombotic microangiopathy. Furthermore, treatment with a beta-blocker apparently improved the outcome of HSCT-GAVE in three patients. PMID- 12900781 TI - Myeloablative therapy and bone marrow transplantation in Jehovah's Witnesses with malignancies: single center experience. AB - Hematological malignancies in Jehovah's Witnesses are often difficult to cure since these patients deny transfusions. By a retrospective analysis, we report the possibility of treating some tumors, mostly hematological, with either autologous or allogeneic bone marrow transplantation (BMT) without blood support. Eight patients were evaluated, including lymphoma (two patients), acute lymphoblastic (one patient) and myeloblastic (one patient) leukemia, chronic lymphocytic leukemia (one patient), refractory anemia with blasts in transformation (one patient), chronic myeloid leukemia (one patient) and metastatic breast cancer (one patient). All patients experienced a severe cytopenia with no major side effects or life-threatening complications. We had four deaths: three from relapse and progression of the disease (at 5, 8 and 15 months after the stem cell infusion), and one from acute intestinal GVHD (at 2 months after the stem cell infusion). Four patients are in complete clinical remission (at 8, 10, 16 and 26 months after the stem cell infusion), and this was related to the disease outcome. We conclude that autologous and allogeneic BMT are feasible without the support of transfusions. We believe that this should be performed as soon as possible in the course of the disease. PMID- 12900780 TI - Monthly prospective analysis of hematopoietic chimerism after allogeneic hematopoietic cell transplantation. AB - SUMMARY: Hematopoietic chimerism (HpC) was assayed monthly using a sensitive, polymerase chain reaction (PCR) -based method in consecutive patients. Between January 1998 and April 2002, 181 patients underwent non-T cell depleted allogeneic hematopoietic cell transplantation (HCT). A total of 163 patients were evaluable for HpC at 1 month (11 early deaths; no informative band for HpC analysis/no genomic DNA in seven). In all, 53 of 163 patients (33%, median recipient DNA of 15% (range 5-95)), 39 of 151 patients (26%), and 27 of 142 patients (19%) showed mixed chimerism (MC) at 1, 2, and 3 months after HCT, respectively. Conditioning regimen (busulfan-fludarabine-ATG vs BuCy, relative risk 3.99 (95% CI 1.16-10.92)), neutrophil engraftment (>/=day 17 vs /=5% recipient DNA at 1 month. Five patients experienced secondary graft failure. All five patients showed MC at 1 month with median recipient DNA of 40%. None of the 109 patients with complete chimerism experienced graft failure (P=0.002). Our study showed that MC shown on monthly analysis of HpC after allogeneic HCT is a significant predictor of secondary graft failure. Bone Marrow Transplantation (2003) 32, 423-431. doi:10.1038/sj.bmt.1704147 PMID- 12900782 TI - Transfusion-free reduced-intensity conditioned allogeneic stem cell transplantation in a Jehovah's witness. AB - With an estimated 6000000 Jehovah's witnesses worldwide, haematologists may encounter patients who decline blood transfusions as a matter of personal belief. Here, we describe the first case of a transfusion-free allogeneic stem cell transplant (SCT) in a Jehovah's witness with CML. The patient underwent HLA identical peripheral blood SCT after reduced-intensity conditioning with total body irradiation and fludarabine. Conditioning and the period after transplantation were uneventful, complete donor chimerism was achieved and the patient continues to be in molecular remission without significant chronic graft versus-host disease. When treating Jehovah's witnesses with diseases that may be cured by allogeneic SCT, this approach should be kept in mind. PMID- 12900783 TI - Induction of graft-versus-autoimmune (GVA) disease effect against refractory psoriasis by complete donor-type chimerism and graft-versus-host disease after allogeneic hematopoietic stem cell transplantation. AB - A 67-year-old man with AML, who had a 21-year history of psoriasis without remission, received a reduced-intensity transplantation from an HLA-identical sibling. The preparative regimen consisted of busulfan and fludarabine. Graft versus-host-disease (GVHD) prophylaxis was cyclosporine and methotrexate. Psoriasis was completely resolved on day 18. The subsequent clinical course was uneventful until day 42, when psoriasis recurred at the same sites as before RIST. Peripheral blood examined on day 63 showed mixed chimerism with 54% recipient type. Cyclosporine was rapidly tapered off over the next 2 weeks. On day 90, 100% donor-type chimerism was confirmed. Subsequently, psoriasis improved simultaneously with the occurrence of mucositis and rash as a manifestation of GVHD. Scattered erythematous patches of psoriasis disappeared again by day 105. We initiated 0.5 mg/kg prednisolone on day 119, and resumed cyclosporine on day 133. At 7 months after RIST, he still suffers from chronic GVHD, but his psoriasis remains in remission for the first time in 21 years. The anti-psoriasis effect of the conditioning is mild and transient, while the graft-versus autoimmunity effect, related to the induction of complete donor-type chimerism and GVHD, is more profound and persisting. A graft-versus-autoimmunity effect lies in the delicate balance between alloimmunity and immunosuppressant used for GVHD prophylaxis/treatment. PMID- 12900784 TI - T-cell-depleted peripheral blood stem cell transplantation for alpha mannosidosis. AB - Alpha-mannosidosis (alpha-mannosidosis) is a lysosomal storage disease characterized by accumulation of oligosaccharides in various tissues leading to symptoms such as coarse facial features, dysostosis multiplex, hearing disabilities, mental developmental delay and skeletal involvement (dysostosis multiplex). Without treatment, the severe infantile onset form of this autosomal recessive disease leads to progressive neurodegeneration and sometimes to early death. Stem cell transplantation has been shown to be an effective treatment. In the five patients published so far, correction of skeletal abnormalities and improvement of neuropsychological capabilities have been observed. We report the first patient who received a T-cell-depleted peripheral blood stem cell transplantation (PBSCT) for alpha-mannosidosis. The diagnosis of alpha mannosidosis was made at the age of 14 months. At the age of 24 months, he underwent PBSCT with T-cell depletion by CD34-positive selection from his HLA phenotypically identical mother. Conditioning was carried out with busulfan (20 mg/kg), cyclophosphamide (200 mg/kg), OKT3 and methylprednisolone. The patient is alive and well 27 months after PBSCT and has made significant developmental progress. The pattern of urinary oligosaccharides has returned to almost normal. CD34-positive-selected PBSCT is a feasible option to reduce risk for GVHD for these patients. PMID- 12900785 TI - Nephrotic syndrome following allogeneic stem cell transplantation associated with increased production of TNF-alpha and interferon-gamma by donor T cells. AB - SUMMARY: Tumor necrosis factor-alpha (TNF-alpha) has been implicated in the immunological complications of stem cell transplantation (SCT) including graft versus-host disease (GVHD). In this report of a patient undergoing allogeneic SCT for AML, serial cytokine measurements by real-time PCR revealed increased production of interferon-gamma (IFN-gamma) and TNF-alpha, but not interleukin (IL)-4 in purified T cells following withdrawal of immunosuppression. Cytokine changes were contemporaneous with the onset of nephrotic syndrome (NS), a rare manifestation of GVHD. These findings indicate that serial cytokine monitoring may allow for the prediction of GVHD during immunosuppression withdrawal and lend further insight into the pathogenesis of NS. Bone Marrow Transplantation (2003) 32, 447-450. doi:10.1038/sj.bmt.1704151 PMID- 12900786 TI - Esophageal actinomycosis after allogeneic peripheral blood stem cell transplantation for extranodal natural killer/T cell lymphoma, nasal type. AB - We report a 19-year-old man with extranodal natural killer (NK)/T cell lymphoma, nasal type treated by allogeneic peripheral blood stem cell transplantation (allo PBSCT). His lymphoma was chemoresistant, and disseminated during local radiotherapy. The patient received allo-PBSCT from his HLA-1 locus mismatched sister using busulfan (BU), cyclophosphamide (CY) and VP-16 as the conditioning regimen. His course was complicated by esophageal actinomycosis 9 months after transplantation, which resulted in the rupture of the right common carotid artery. These observations suggest that actinomycosis should be monitored carefully after transplantation in patients who have received local radiation therapy before the procedure. PMID- 12900787 TI - The meaning of psychosocial occupational therapy in a life-story perspective. A long-term follow-up of three cases. AB - This study is a long-term follow-up in narrative form of three former psychiatric patients who had been discharged eight to nine years earlier from an outpatient occupational therapy programme. The purpose of the study was to gain an understanding about the participants' views about the period of therapy from a long-term perspective and of how the outcome of therapy could be understood within the context of the patients' life plots. In-depth retrospective interviews and two former interviews with each participant, conducted at the time of therapy, comprised the data. The interview transcripts were subjected to narrative analysis. The fit between the characteristics of the therapy programme and important life themes of the informants seemed crucial for the long-term outcome of therapy, suggesting that taking life histories is important for occupational therapists in clinical practice in order to design appropriate interventions. Four components of the therapy, the activities, the social interaction, the milieu, and the therapeutic relationship, appeared as vital in re-shaping the informants' life plots. PMID- 12900788 TI - Facilitating professional identify in occupational therapy students. AB - The purpose of this study was to explore how a graduate course in occupational therapy theory can help prepare students to develop a professional identity. Thirty students participated in the study. The course included content on the history of occupational therapy, models of practice and the social, philosophical, political and economic factors that impact on occupational therapy. The students were divided into groups of four or five where they discussed issues assigned by the instructor. An electronic blackboard was used to share the discussion with the class. Surveys of the students' opinions were used to collect data on what the students viewed as the strengths and weaknesses of the course. The students felt that the class discussions were the strongest part of the course. They felt that the course improved their critical thinking and problem solving significantly. It was concluded from the results that the course facilitated their professional identity through the connection made between theory and practice. There is a recognized need to explore the issue of developing a professional identity in occupational therapy students through courses integrating philosophical topics and clinical practice. Specifically, it is recommended that further research be carried out in educational settings with larger samples, using comparison groups and other methodologies to evaluate the issue of facilitating professional identity in occupational therapy students. PMID- 12900789 TI - Stability of tactile defensiveness across cultures: European and American children's responses to the Touch Inventory for Elementary school aged children. AB - Screening and assessment for single and discrete sensory systems, such as screening for tactile defensiveness, can provide valid information for identifying sensory processing dysfunction of children. Tactile defensiveness may be considered to be a specialized type of what was previously called a sensory integrative disorder and now is termed a sensory modulation disorder. The current study was designed to examine further the phenomenon of tactile defensiveness, particularly the stability of the concept, across different cultures. In this study, the test scores of 28 European children (16 girls and 12 boys of three different nationalities) on the Touch Inventory for Elementary School Aged Children (TIE) were collected and analysed (Royeen and Fortune, 1990). The TIE scores of the European sample were compared with that of the American sample in the original TIE study (Royeen and Fortune, 1990). Results revealed that the mean test scores on the TIE between the American sample and European sample were very similar (n = 415, M = 41.0 vs. n = 28, M = 39.5) and no significant difference was found between the two samples (t = 0.99, p < 0.05). Internal consistency of the TIE on the European sample was found to be good and close to that obtained from the American sample (0.78 vs. 0.89). The current study provides evidence that tactile defensiveness is a stable phenomenon across different cultures as measured by the TIE. Limitations of the study include use of a small and convenient sample of European children. Further studies are recommended to examine tactile defensiveness as a clinical phenomenon. PMID- 12900790 TI - Treatment of sensory defensiveness in adults. AB - This pilot study explored the relationship between sensory defensiveness and anxiety, as well as the impact of a sensory integration treatment protocol on normal adults. Fifteen adult subjects identified as having sensory defensiveness completed the Adult Sensory Questionnaire (ASQ), Adult Sensory Interview (ADULT SI), and Beck Anxiety Inventory (BAI) at pre-test and post-test intervals to measure sensory defensiveness and anxiety. A treatment protocol was implemented which included providing insight into sensory defensiveness, regular and daily sensory input, and engagement in activities of choice providing primarily proprioceptive, vestibular and tactile sensory input. Subjects engaged in an individualized self-treatment protocol for one month. Results indicated a significant correlation between anxiety and sensory defensiveness (r = 0.62, p = 0.027). The differences in pre-test and post-test mean scores of the Adult Sensory Interview (p = 0.048) and the Beck Anxiety Inventory (p = 0.0453) supported the use of a sensory treatment protocol to decrease sensory defensiveness and secondary anxiety. There is a need for further research using experimental methodology with a control group to explore the effectiveness of a sensory treatment protocol on sensory defensiveness and anxiety. PMID- 12900791 TI - The paternal-age effect in Apert syndrome is due, in part, to the increased frequency of mutations in sperm. AB - A paternal-age effect and the exclusive paternal origin of mutations have been reported in Apert syndrome (AS). As the incidence of sporadic AS births increases exponentially with paternal age, we hypothesized that the frequency of AS mutations in sperm would also increase. To determine the frequency of two common FGFR2 mutations in AS, we developed allele-specific peptide nucleic acid-PCR assays. Analyzing sperm DNA from 148 men, age 21-80 years, we showed that the number of sperm with mutations increased in the oldest age groups among men who did not have a child with AS. These older men were also more likely to have both mutations in their sperm. However, this age-related increase in mutation frequency was not sufficient to explain the AS-birth frequency. In contrast, the mutation frequency observed in men who were younger and had children with AS was significantly greater. In addition, our data suggest selection for sperm with specific mutations. Therefore, contributing factors to the paternal-age effect may include selection and a higher number of mutant sperm in a subset of men ascertained because they had a child with AS. No age-related increase in the frequency of these mutations was observed in leukocytes. Selection and/or quality control mechanisms, including DNA repair and apoptosis, may contribute to the cell-type differences in mutation frequency. PMID- 12900792 TI - A genome scan for modifiers of age at onset in Huntington disease: The HD MAPS study. AB - Huntington disease (HD) is caused by the expansion of a CAG repeat within the coding region of a novel gene on 4p16.3. Although the variation in age at onset is partly explained by the size of the expanded repeat, the unexplained variation in age at onset is strongly heritable (h2=0.56), which suggests that other genes modify the age at onset of HD. To identify these modifier loci, we performed a 10 cM density genomewide scan in 629 affected sibling pairs (295 pedigrees and 695 individuals), using ages at onset adjusted for the expanded and normal CAG repeat sizes. Because all those studied were HD affected, estimates of allele sharing identical by descent at and around the HD locus were adjusted by a positionally weighted method to correct for the increased allele sharing at 4p. Suggestive evidence for linkage was found at 4p16 (LOD=1.93), 6p21-23 (LOD=2.29), and 6q24 26 (LOD=2.28), which may be useful for investigation of genes that modify age at onset of HD. PMID- 12900793 TI - Estimation of the inbreeding coefficient through use of genomic data. AB - Many linkage studies are performed in inbred populations, either small isolated populations or large populations with a long tradition of marriages between relatives. In such populations, there exist very complex genealogies with unknown loops. Therefore, the true inbreeding coefficient of an individual is often unknown. Good estimators of the inbreeding coefficient (f) are important, since it has been shown that underestimation of f may lead to false linkage conclusions. When an individual is genotyped for markers spanning the whole genome, it should be possible to use this genomic information to estimate that individual's f. To do so, we propose a maximum-likelihood method that takes marker dependencies into account through a hidden Markov model. This methodology also allows us to infer the full probability distribution of the identity-by descent (IBD) status of the two alleles of an individual at each marker along the genome (posterior IBD probabilities) and provides a variance for the estimates. We simulate a full genome scan mimicking the true autosomal genome for (1) a first-cousin pedigree and (2) a quadruple-second-cousin pedigree. In both cases, we find that our method accurately estimates f for different marker maps. We also find that the proportion of genome IBD in an individual with a given genealogy is very variable. The approach is illustrated with data from a study of demyelinating autosomal recessive Charcot-Marie-Tooth disease. PMID- 12900794 TI - A full-likelihood method for the evaluation of causality of sequence variants from family data. AB - In many disease genes, a substantial fraction of all rare variants detected cannot yet be used for genetic counselling because of uncertainty about their association with disease. One approach to the characterization of these unclassified variants is the analysis of patterns of cosegregation with disease in affected carrier families. Petersen et al. previously provided a simplistic Bayesian method for evaluation of causality of such sequence variants. In the present report, we propose a more general method based on the full pedigree likelihood, and we show that the use of this method can provide more accurate and informative assessment of causality than could the previous method. We further show that it is important that the pedigree information be as complete as possible and that the distinction be made between unaffected individuals and those of unknown phenotype. PMID- 12900796 TI - Selection and evaluation of tagging SNPs in the neuronal-sodium-channel gene SCN1A: implications for linkage-disequilibrium gene mapping. AB - Association studies are widely seen as the most promising approach for finding polymorphisms that influence genetically complex traits, such as common diseases and responses to their treatment. Considerable interest has therefore recently focused on the development of methods that efficiently screen genomic regions or whole genomes for gene variants associated with complex phenotypes. One key element in this search is the use of linkage disequilibrium to gain maximal information from typing a selected subset of highly informative single-nucleotide polymorphism (SNP) markers, now often called "tagging SNPs" (tSNPs). Probably the most common approach to linkage-disequilibrium gene mapping involves a three-step program: (1) characterization of the haplotype structure in candidate genes or genomic regions of interest, (2) identification of tSNPs sufficient to represent the most common haplotypes, and (3) typing of tSNPs in clinical material. Early definitions of tSNPs focused on the amount of haplotype diversity that they explained. To select tSNPs that would have maximal power in a genetic association study, however, we have developed optimization criteria based on the r2 measure of association and have compared these with other criteria based on the haplotype diversity. To evaluate the full program and to assess how well the selected tags are likely to perform, we have determined the haplotype structure and have assessed tSNPs in the SCN1A gene, an important candidate gene for sporadic epilepsy. We find that as few as four tSNPs are predicted to maintain a consistently high r2 value with all other common SNPs in the gene, indicating that the tags could be used in an association study with only a modest reduction in power relative to direct assays of all common SNPs. This implies that very large case-control studies can be screened for variation in hundreds of candidate genes with manageable experimental effort, once tSNPs are identified. However, our results also show that tSNPs identified in one population may not necessarily perform well in another, indicating that the preliminary study to identify tSNPs and the later case-control study should be performed in the same population. Our results also indicate that tSNPs will not easily identify discrepant SNPs, which lie on importantly discriminating but apparently short genealogical branches. This could significantly complicate tagging approaches for phenotypes influenced by variants that have experienced positive selection. PMID- 12900797 TI - Age-related macular degeneration--a genome scan in extended families. AB - We performed a genomewide scan and genetic linkage analysis, to identify loci associated with age-related macular degeneration (AMD). We collected 70 families, ranging from small nuclear families to extended multigenerational pedigrees and consisting of a total of 344 affected and 217 unaffected members available for genotyping. We performed linkage analyses using parametric and allele-sharing models. We performed the analyses on the complete pedigrees but also subdivided the families into nuclear pedigrees. Finally, to dissect potential genetic factors responsible for differences in disease manifestation, we stratified the sample by two major AMD phenotypes (neovascular AMD and geographic atrophy) and by age of affected family members at the time of our evaluation. We have previously demonstrated linkage between AMD and 1q25-31 in a single large family. In the combined sample, we have detected the following loci with scores exceeding a LOD=2 cutoff under at least one of the models considered: 1q31 (HLOD=2.07 at D1S518), 3p13 (HLOD=2.19 at D3S1304/D3S4545), 4q32 (HLOD=2.66 at D4S2368, for the subset of families with predominantly dry AMD), 9q33 (LODZlr=2.01 at D9S930/D9S934), and 10q26 (HLOD=3.06 at D10S1230). Using correlation analysis, we have found a statistically significant correlation between LOD scores at 3p13 and 10q26, providing evidence for epistatic interactions between the loci and, hence, a complex basis of AMD. Our study has identified new loci that should be considered in future mapping and mutational analyses of AMD and has strengthened the evidence in support of loci suggested by other studies. PMID- 12900795 TI - Genetic disorders of the skeleton: a developmental approach. AB - Although disorders of the skeleton are individually rare, they are of clinical relevance because of their overall frequency. Many attempts have been made in the past to identify disease groups in order to facilitate diagnosis and to draw conclusions about possible underlying pathomechanisms. Traditionally, skeletal disorders have been subdivided into dysostoses, defined as malformations of individual bones or groups of bones, and osteochondrodysplasias, defined as developmental disorders of chondro-osseous tissue. In light of the recent advances in molecular genetics, however, many phenotypically similar skeletal diseases comprising the classical categories turned out not to be based on defects in common genes or physiological pathways. In this article, we present a classification based on a combination of molecular pathology and embryology, taking into account the importance of development for the understanding of bone diseases. PMID- 12900801 TI - Analgesia in patients with ESRD: a review of available evidence. AB - Moderate to severe pain frequently accompanies chronic diseases in general and end-stage renal disease (ESRD) in particular. Several analgesic agents and associated metabolites show altered pharmacokinetics in the presence of reduced glomerular filtration rate. Drug-related side effects may exacerbate symptoms frequently observed in persons with chronic kidney disease (CKD; eg, fatigue, nausea, vomiting, and constipation) or those often attributed to hemodialysis therapy (eg, orthostatic hypotension and impaired cognition). Persons with advanced CKD and ESRD are at increased risk for adverse effects of analgesic agents because of enhanced drug sensitivity, comorbid conditions, and concurrent medication use. Dose adjustment and avoidance of certain analgesics may be required in patients with advanced CKD and ESRD. We review the available evidence on pharmacokinetics and adverse drug effects of various analgesic agents commonly used in patients with advanced CKD and ESRD. Determining an optimal approach to the control of pain in patients with advanced CKD and ESRD will require additional research. PMID- 12900798 TI - Y-chromosome evidence for differing ancient demographic histories in the Americas. AB - To scrutinize the male ancestry of extant Native American populations, we examined eight biallelic and six microsatellite polymorphisms from the nonrecombining portion of the Y chromosome, in 438 individuals from 24 Native American populations (1 Na Dene and 23 South Amerinds) and in 404 Mongolians. One of the biallelic markers typed is a recently identified mutation (M242) characterizing a novel founder Native American haplogroup. The distribution, relatedness, and diversity of Y lineages in Native Americans indicate a differentiated male ancestry for populations from North and South America, strongly supporting a diverse demographic history for populations from these areas. These data are consistent with the occurrence of two major male migrations from southern/central Siberia to the Americas (with the second migration being restricted to North America) and a shared ancestry in central Asia for some of the initial migrants to Europe and the Americas. The microsatellite diversity and distribution of a Y lineage specific to South America (Q-M19) indicates that certain Amerind populations have been isolated since the initial colonization of the region, suggesting an early onset for tribalization of Native Americans. Age estimates based on Y-chromosome microsatellite diversity place the initial settlement of the American continent at approximately 14,000 years ago, in relative agreement with the age of well-established archaeological evidence. PMID- 12900802 TI - Developing a comprehensive diagnostic and interventional nephrology program at an academic center. AB - Procedure-related delays in the treatment of patients with renal disease can be minimized and nephrology care can be delivered more efficiently by a nephrologist trained in nephrology-related procedures. Referrals to a radiologist for renal ultrasound and biopsy, to a surgeon for dialysis access placement, and to an interventional radiologist for dialysis catheter placement and vascular access procedures are time consuming and inconvenient to patients with renal disease. Moreover, such an approach may result in delays in the availability of critical diagnostic information and a break in the continuity of care. In an effort to optimize the care of nephrology patients, we developed a diagnostic and interventional nephrology (DIN) program that effectively deals with nephrology related procedures in a timely manner. At present, some of the commonly performed nephrology-related procedures at our center include diagnostic ultrasonography, ultrasound-guided renal biopsy, peritoneal dialysis access procedures, permanent tunneled hemodialysis catheter placement, and endovascular procedures for arteriovenous dialysis access dysfunction. To date, we have performed 893 procedures during a period of 2 years. This article describes our approach and the tools required to develop a DIN program at an academic medical center. PMID- 12900803 TI - Analgesic use and change in kidney function in apparently healthy men. AB - BACKGROUND: Studies using a single creatinine measurement have yielded inconsistent results regarding analgesic use and kidney function. METHODS: This is a prospective cohort study of 4,494 US male physicians who provided blood samples in both 1982 and 1996. Outcomes measured were increase in plasma creatinine level of 0.3 mg/dL or greater (> or =26.5 micromol/L) and decline in glomerular filtration rate (GFR) of 29.0 mL/min/1.73 m2 or greater during this 14 year period. Self-reported use of aspirin, acetaminophen, and other nonsteroidal anti-inflammatory drugs (NSAIDs) was classified as never (<12 pills during the study period), 12 to 1,499 pills, 1,500 to 2,499 pills, and 2,500 or greater pills during the study period. RESULTS: Blood measurements made 14 years apart showed increased creatinine levels in 242 participants and decreased GFRs in 224 participants. Compared with never use, multivariable-adjusted odds ratios (ORs) of increased creatinine levels were 0.98 (95% confidence interval [CI], 0.43 to 2.23) for 2,500 or greater pills of aspirin, 1.02 (95% CI, 0.55 to 1.90) for 2,500 or greater pills of acetaminophen, and 1.12 (95% CI, 0.67 to 1.87) for 2,500 or greater pills of other NSAIDs. For decreased GFRs, ORs for intake of 2,500 or greater pills were 0.75 (95% CI, 0.35 to 1.57) for aspirin, 1.22 (95% CI, 0.66 to 2.26) for acetaminophen, and 1.11 (95% CI, 0.65 to 1.90) for other NSAIDs. Use of aspirin, but not acetaminophen or other NSAIDs, was associated with a reduced risk for change in kidney function in participants without cardiovascular risk factors and a possible but nonsignificant increase in those with cardiovascular risk factors. CONCLUSION: Occasional to moderate analgesic intake of aspirin, acetaminophen, or NSAIDs does not appear to increase the risk for decline in kidney function during a period of 14 years. PMID- 12900804 TI - Testing for microalbuminuria in 2002: barriers to implementing current guidelines. AB - BACKGROUND: Testing for microalbuminuria is recommended to detect early kidney damage in patients with diabetes or other diseases. However, few studies have examined laboratory practices for microalbuminuria testing in the general community. METHODS: In 2002, all laboratories in Montana and reference laboratories used by Montana laboratories for microalbuminuria measurement were surveyed by mail to ascertain if they provided testing for microalbuminuria, specific tests performed, and units and cutoff values used to report microalbuminuria results. RESULTS: One hundred three of 126 laboratories (82%) responded to the survey. Overall, 79% of laboratories offered quantitative testing for microalbuminuria, either on site or through a reference laboratory. Twenty-five laboratories (24%) surveyed provided quantitative testing for microalbuminuria on site. Only 14 of 23 laboratories offering albumin-creatinine ratios on site reported results in units and cutoff values consistent with current recommendations. Fewer laboratories provided 24-hour (6 of 17 laboratories) or other timed (2 of 7 laboratories) testing, and many of these laboratories did not report results using recommended units and cutoff values. Overall, only 11 of 25 laboratories (44%) with on-site testing reported microalbuminuria values from 1 or more types of specimens exclusively using recommended units and cutoff values. CONCLUSION: Quantitative testing for microalbuminuria is not offered universally, and results often are reported in units and cutoff values that differ from current clinical recommendations. PMID- 12900805 TI - Receipt of renal replacement therapy in the United States: a population-based study of sociodemographic disparities from the Second National Health and Nutrition Examination Survey (NHANES II). AB - BACKGROUND: Persons with chronic kidney disease who need kidney replacement therapy to sustain life have health insurance. We examined whether young adults, women, blacks, less-educated persons, the poor, and persons residing in less populated areas receive treatment when health insurance is no longer a barrier. METHODS: We conducted a case-control study nested in the Second National Health and Nutrition Examination Survey Mortality Study. Cases were persons treated with kidney replacement therapy determined by linkage to the end-stage renal disease treatment registry. Controls were untreated persons with kidney disease who died not appearing in the registry. RESULTS: During 12 to 16 years, 44 persons developed treated disease, and 145 persons, untreated disease. After adjustment for sex, age, education, population of residential area, and comorbid conditions in logistic regression analysis, younger versus older age and living in a highly populated versus less populated area were both independently associated with treatment (relative odds of treatment, 5.57; 95% confidence interval, 1.72 to 18.0; and 4.33; 95% confidence interval, 2.09 to 8.97, respectively). Race, sex, education, and poverty were not associated with less treatment. CONCLUSION: We found no disparity in life-saving chronic kidney disease treatment with regard to race or socioeconomic status in this population-based study. Less receipt of treatment by older adults may reflect greater comorbid disease or choices made by persons or their providers. Strategies to render treatment in less populated areas, including incentives to deliver care to such areas, should be encouraged. PMID- 12900806 TI - Leukocytosis, hypoalbuminemia, and the risk for chronic kidney disease in US adults. AB - BACKGROUND: Atherosclerosis and chronic kidney disease (CKD) share several common antecedents. However, the association between inflammatory markers and incident CKD is unknown. METHODS: We determined risk for incident CKD, defined by treatment for kidney failure or death related to kidney disease, in 9,250 US adults aged 30 to 74 years who participated in the Second National Health and Nutrition Examination Survey (NHANES II), a nationally representative prospective cohort study with 17 years of follow-up. RESULTS: After adjusting for age, race, sex, blood pressure, smoking, and body mass index, there was a graded positive association with increasing total white blood cell (WBC) count and risk for CKD (P for trend < 0.001; relative hazard (RH) highest versus lowest quartile, 2.34; 95% confidence interval [CI], 1.30 to 4.19). This association remained statistically significant after adjusting further for the presence of diabetes and cardiovascular disease at baseline (RH, 2.01; 95% CI, 1.11 to 3.65). A similarly strong and graded association with incident CKD was observed for hypoalbuminemia after adjusting for age, race, sex, blood pressure, smoking, and body mass index (P for trend = 0.02; RH lowest versus highest quartile, 1.91; 95% CI, 0.89 to 4.07) and additionally adjusting for the presence of diabetes and cardiovascular disease at baseline (P for trend = 0.02; RH lowest versus highest, 2.05; 95% CI, 0.96 to 4.39). CONCLUSION: In a nationally representative sample of US adults, elevated WBC count and hypoalbuminemia were associated with future risk for CKD. These results support the hypothesis that systemic inflammation is an independent risk factor for CKD. PMID- 12900807 TI - Effects of pentoxifylline administration on urinary N-acetyl-beta-glucosaminidase excretion in type 2 diabetic patients: a short-term, prospective, randomized study. AB - BACKGROUND: Tubulointerstitial injury is a major feature of diabetic nephropathy and an important predictor of renal dysfunction. In 45 patients with type 2 diabetes mellitus (DM), we prospectively analyzed urinary excretion of N-acetyl beta-glucosaminidase (NAG), a marker of tubular renal damage; the potential relationship with urinary protein excretion; and effects of pentoxifylline (PTF) administration. METHODS: Forty-five patients with type 2 DM initially were compared with 15 healthy controls matched for age and sex. After randomization, PTF (1,200 mg/d) was administered for 4 months to 30 patients and results were compared with data from a control group (n = 15). RESULTS: Proteinuria and urinary NAG excretion were significantly greater in patients with DM with respect to healthy controls. Before PTF administration, baseline parameters were similar in both groups of patients with DM. At the end of the study, urinary protein excretion and NAG-creatinine ratios decreased in the active group from 920 +/- 522 mg/d and 14.3 +/- 16.9 U/g to 803 +/- 523 mg/d (P < 0.001) and 10.5 +/- 9.3 U/g (P < 0.05), respectively. Conversely, proteinuria and urinary NAG excretion did not change in the control group. Regression analysis showed that urinary NAG excretion was significantly associated with duration of DM (r = 0.61; P < 0.001) and proteinuria (r = 0.51; P < 0.001). CONCLUSION: Urinary NAG excretion is elevated in patients with type 2 DM compared with healthy individuals and increases as nephropathy progresses. PTF administration is effective in reducing proteinuria and urinary NAG excretion in these patients. These findings suggest that PTF may have beneficial effects on tubulointerstitial damage in diabetic kidney disease. PMID- 12900808 TI - Polymorphisms of interleukin-4--related genes in Japanese children with minimal change nephrotic syndrome. AB - BACKGROUND: Minimal change nephrotic syndrome (MCNS) in children frequently is associated with allergy and immunoglobulin E production. T Helper subtype 2 cytokines, such as interleukin-4 (IL-4), may have an important role in the development of atopy. We investigated the associations of gene polymorphisms of IL-4, its receptor (IL-4R), and the signal transducer and activator 6 (STAT6) gene with MCNS. METHODS: We analyzed these polymorphisms in Japanese children with MCNS (n = 58) and healthy controls with neither allergic nor renal disease (n = 63). The polymerase chain reaction (PCR) and restriction fragment length polymorphism method was used for the IL-4 promoter gene polymorphism (-590C/T), and PCR single-strand conformation polymorphism analysis was used for the IL 4Ralpha chain gene (1902A/G) and STAT6 3' untranslated region (2964G/A) polymorphisms. RESULTS: There was a significant difference between the MCNS group and controls in genotypic distribution of IL-4 promoter gene polymorphism. Frequency of the T allele was significantly lower in the MCNS group than controls. There was no difference between the MCNS group and controls in the IL 4R gene polymorphism. In the STAT6 gene, no significant differences in genotypic and allelic distribution were observed between the 2 groups. However, there were significant differences between patients who did not need cytotoxic agents or who experienced 3 or fewer relapses and controls. CONCLUSION: These results suggest that genetic variations in the IL-4 and STAT6 genes may be associated with predisposition to MCNS, partially the clinical course of MCNS. PMID- 12900809 TI - Risk factors for aortic atherosclerosis determined by transesophageal echocardiography in patients with CRF. AB - BACKGROUND: The significance of various risk factors for cardiovascular disease (CVD) in the pathogenesis of atherosclerosis in patients with chronic renal failure (CRF) is, to a great deal, unresolved. The high risk for CVD in patients with CRF may be caused by the high prevalence of recognized risk factors for CVD or by factors characteristic of CRF in these patients. In this prospective cross sectional study, we examined risk factors for thoracic aortic atherosclerosis in a population of patients with CRF consisting of predialysis and dialysis patients, as well as renal transplant recipients. METHODS: Of 118 patients, 52 patients had moderate to severe predialysis CRF, 32 patients were on dialysis treatment, and 34 patients were renal transplant recipients. Mean age was 52 +/- 12 years, and 35 patients (30%) had diabetes. Multiplane transesophageal echocardiography (TEE) was performed using local anesthesia. RESULTS: Large aortic plaques (LAPs; > or = 3.0 mm in diameter) were found in 39 patients (33%). In univariate analysis, age, duration of hypertension, pulse pressure, low diastolic blood pressure, elevated fibrinogen level, C-reactive protein level, total cholesterol level, low-density lipoprotein cholesterol level, and duration of dialysis or a functioning renal transplant were significantly associated (P < 0.05) with LAP. In multivariate analysis, age, duration of hypertension, and total cholesterol level were associated with LAP. CONCLUSION: Results of the present TEE study suggest that in addition to duration of hypertension and renal disease, hypercholesterolemia has a role in the pathogenesis of atherosclerosis in patients with CRF. PMID- 12900810 TI - Linkage of hypoalbuminemia, inflammation, and oxidative stress in patients receiving maintenance hemodialysis therapy. AB - BACKGROUND: Hypoalbuminemia is a powerful predictor of cardiovascular mortality in maintenance hemodialysis patients. Increased biomarkers of acute-phase inflammation and oxidative stress are highly prevalent and also correlate with cardiovascular morbidity and mortality. The extent to which hypoalbuminemia, biomarkers of inflammation, and biomarkers of oxidative stress are linked in this patient population is unknown. We hypothesized that a high proportion of hypoalbuminemic hemodialysis patients also would manifest increased levels of biomarkers of inflammation and oxidative stress. METHODS: We surveyed 600 maintenance hemodialysis patients and identified 18 severely hypoalbuminemic patients (serum albumin level < 3.2 g/dL [32 g/L]) without recent infection or hospitalization. We then identified 18 age-, race-, sex-, and diabetes-matched normoalbuminemic hemodialysis patients, as well as 18 age-, race-, sex-, and diabetes-matched healthy subjects, for cohort comparison. Measurements of plasma interleukin-6 (IL-6) levels, plasma protein reduced thiol content, plasma protein carbonyl content, and plasma free F2-isoprostane levels, as well as serum concentrations of C-reactive protein (CRP) and prealbumin, were performed for study purposes. RESULTS: Levels of serum CRP, IL-6, plasma protein thiol oxidation, and protein carbonyl formation were significantly elevated in both hypoalbuminemic and normoalbuminemic hemodialysis patients compared with healthy subjects and also were significantly different in hypoalbuminemic maintenance dialysis patients compared with normoalbuminemic hemodialysis patients. Prealbumin levels were significantly lower in hypoalbuminemic hemodialysis patients than in other groups. CONCLUSION: There is a high prevalence of inflammation and oxidative stress in the maintenance hemodialysis population. Levels of inflammatory and oxidative stress biomarkers are increased further in hypoalbuminemic compared with normoalbuminemic dialysis patients. Hypoalbuminemia, acute-phase inflammation, and oxidative stress may act synergistically to increase cardiovascular morbidity and mortality risk in maintenance hemodialysis patients. PMID- 12900811 TI - Association between inflammatory mediators and muscle mass in long-term hemodialysis patients. AB - BACKGROUND: Muscle wasting is highly prevalent in long-term hemodialysis (HD) patients. Although inflammatory indices have been associated with malnutrition in these patients, the role of inflammation in muscle wasting has not yet been determined. METHODS: The relationship between the inflammatory mediators C reactive protein (CRP) and interleukin-6 (IL-6) and the muscle mass indices thigh muscle area (TMA), measured by computed tomography, and creatinine (Cr) production, estimated by the Cr kinetic model (Cr-CKM), were investigated in 188 HD patients. RESULTS: Serum IL-6 level (7.3 +/- 7.8 pg/mL) was significantly elevated in HD patients, whereas mean serum CRP level (4.8 +/- 7.5 mg/L) remained within the normal range. Similar to serum albumin, muscle mass indices had significantly negative correlations with both serum IL-6 and CRP levels (TMA/dry weight [DW] versus log IL-6, r = -0.28; P < 0.01; TMA/DW versus log CRP, r = 0.38; P < 0.001; Cr-CKM versus log IL-6, r = -0.31; P < 0.01; Cr-CKM versus log CRP, r = -0.24; P < 0.01). Although muscle mass indices also were associated with both age and sex, a multiple regression analysis confirmed that these inflammatory indices were significantly associated with muscle mass in HD patients. CONCLUSION: Data indicate that muscle wasting is associated closely with inflammatory indices in long-term HD patients. It may be important to clarify the mechanism for the increasing inflammatory status and suppress the inflammatory response in these patients to improve their malnutrition and recover muscle mass. PMID- 12900812 TI - The progression of vascular calcification and serum osteoprotegerin levels in patients on long-term hemodialysis. AB - BACKGROUND: The aortic calcification index (ACI), estimated on abdominal computed tomographic scans, has been associated with the extent of arteriosclerosis in hemodialysis patients. However, the contribution of biochemical markers to the progression of vascular calcification in patients undergoing hemodialysis is not fully understood. METHODS: We examined the relationship between coronary risk factors; metabolic factors, including serum osteoprotegerin (OPG) concentration; and progression of vascular calcification in 26 dialysis patients. RESULTS: Mean patient age was 52.6 +/- 8.7 (SD) years, and mean duration of dialysis therapy was 7.7 +/- 5.8 years. ACI was measured twice in each patient, and the mean interscan period was 4.9 +/- 0.3 years. Mean ACI changed from 22.2 +/- 24.2 to 33.9 +/- 28.8 overall, and mean change in ACI (DeltaACI) was 12.0 +/- 9.9. Patients were divided into 2 groups: slow progressors, with DeltaACI of 4.1 +/- 3.2 (n = 13), and rapid progressors, with DeltaACI of 19.8 +/- 7.9 (n = 13). Serum fasting glucose and CRP levels of rapid progressors were high, and their serum albumin and intact parathyroid hormone levels were low. Multiple regression analyses showed that serum OPG levels were independently associated with vascular calcification in the hemodialysis patients studied. CONCLUSION: Rapid progression of vascular calcification was associated with dose of calcium carbonate prescribed and serum OPG concentration. The clinical significance of these observations remains to be determined. PMID- 12900813 TI - Single-dose pharmacokinetics of ofloxacin during continuous venovenous hemofiltration in critical care patients. AB - BACKGROUND: Ofloxacin is a quinolone administered to patients with severe infections. Pharmacokinetic data on ofloxacin in critically ill patients on renal replacement therapy are sparse and conflicting. METHODS: Eight patients with anuric acute renal failure were administered 400 mg of ofloxacin intravenously. The pharmacokinetics of ofloxacin was analyzed during continuous venovenous hemofiltration (CVVH) with a high-flux polysulfone membrane. Serum and ultrafiltrate levels of ofloxacin were measured by means of high-performance liquid chromatography. RESULTS: Mean serum ofloxacin concentration peak was 5.5 +/- 0.7 mg/L. Elimination half-life, hemofiltration clearance, and total removal were 2.8 +/- 0.5 hours, 89.9 +/- 4.5 mL/min, and 76.9% +/- 7.1%, respectively. CONCLUSION: Ofloxacin is significantly and rapidly eliminated by CVVH with a high flux polysulfone membrane. PMID- 12900814 TI - A multicenter, prospective, randomized, comparative evaluation of dual- versus triple-lumen catheters for hemodialysis and apheresis in 485 patients. AB - BACKGROUND: The purpose of this study is to compare a new temporary triple-lumen catheter (TLC) for dialysis that has a third lumen devoted to fluid and medication administration or blood sampling with a marketed dual-lumen catheter (DLC). METHODS: Four hundred eighty-five patients referred for acute hemodialysis or apheresis were randomly assigned to either a TLC or DLC in a multicenter, prospective, randomized trial. RESULTS: Analysis of blood flow rates was completed on 464 patients (228 patients, DLC; 236 patients, TLC) with a total of 1,681 hemodialysis (808 treatments, DLC; 873 treatments, TLC) and 82 apheresis treatments (37 treatments, DLC; 45 treatments, TLC). During hemodialysis, a median achieved flow rate (AFR) of 267 mL/min was realized for both groups (P = 0.58). During apheresis, a median AFR of 72.5 mL/min (range, 50 to 150 mL/min) was achieved in the DLC group, and 87 mL/min (range, 60 to 150 mL/min), in the TLC group (P = 0.14). Three hundred ninety-three patients (193 patients, DLC; 200 patients, TLC) had blood and catheter tip cultures performed on removal, and catheter-related bloodstream infection (CRBSI) status was determined. Thirty-one patients (7.9%) had a CRBSI: 16 patients (8.3%), DLC; and 15 patients (7.5%), TLC (P= 0.77). Incidence densities of CRBSI were 12.4/1,000 DLC-days and 10.2/1,000 TLC-days (P = 0.59). The CRBSI incidence of 18.2/1,000 catheter-days for femoral sites was significantly greater than the 7/1,000 catheter-days for jugular sites (P = 0.02) and 6.6/1,000 catheter-days for combined jugular and subclavian sites (P = 0.01). In multivariate analysis, antibiotic use was the only factor related to CRBSI (odds ratio, 0.30; 95% confidence interval, 0.12 to 0.76). There were no statistically significant differences in rates of other complications between the 2 catheters. CONCLUSION: Results show that the new TLC is similar to the marketed DLC. PMID- 12900816 TI - Diagnostic accuracy of ultrasound dilution access blood flow measurement in detecting stenosis and predicting thrombosis in native forearm arteriovenous fistulae for hemodialysis. AB - BACKGROUND: Vascular access surveillance by ultrasound dilution blood flow rate (Qa) measurement is widely recommended; however, optimal criteria for detecting stenosis and predicting thrombosis in arteriovenous fistulae (AVFs) are still not clearly defined. METHODS: In a blinded trial, we evaluated the accuracy of single Qa measurement, Qa adjusted for mean arterial pressure (Qa/MAP), and decrease in Qa over time (dQa) in detecting stenosis and predicting thrombosis in an unselected population of 120 hemodialysis subjects with native forearm AVFs (91 AVFs, located at the wrist; 29 AVFs, located at the midforearm). All AVFs underwent fistulography, which identified greater than 50% stenosis in 54 cases. RESULTS: Receiver operating characteristic curve analysis showed that dQa, Qa, and Qa/MAP have a high stenosis discriminative ability with similar areas under the curve (AUCs), ie, 0.961 +/- 0.025, 0.946 +/- 0.021, and 0.912 +/- 0.032, respectively. In the population as a whole, optimal thresholds for stenosis were Qa less than 750 mL/min alone and in combination with dQa greater than 25% (efficiency, 90%); however, the best threshold depended on anastomotic site; it was Qa less than 750 mL/min for an AVF at the wrist and Qa less than 1,000 mL/min for an AVF in the midforearm. Qa was the best predictor of incipient thrombosis (AUC, 0.981 +/- 0.013) with an optimal threshold at less than 300 mL/min (efficiency, 94%). Pooled intra-assay and interassay variation coefficients were 8.2% for MAP, 7.9% for Qa, and 11.2% for Qa/MAP. CONCLUSION: Our study shows that ultrasound dilution Qa measurement is a reproducible and highly accurate tool for detecting stenosis and predicting thrombosis in forearm AVFs. Neither Qa/MAP nor dQa improve the diagnostic performance of Qa alone, although its combination with dQa increases the test's sensitivity for stenosis. PMID- 12900815 TI - Clinical evaluation of heme iron polypeptide: sustaining a response to rHuEPO in hemodialysis patients. AB - BACKGROUND: Optimizing iron and recombinant human erythropoietin (rHuEPO) therapy is necessary to achieve target hemoglobin levels and minimize costs as the end stage renal disease (ESRD) population expands. Oral iron products in patients with ESRD have been largely abandoned, and the safety of intravenous (IV) iron preparations has improved with the introduction of new-generation compounds that have little allergenicity. Recent work suggests oral heme iron may be an effective supplement for hemodialysis (HD) patients because it is absorbed by patients with high ferritin levels, has fewer side effects, and its absorption is stimulated by erythropoietin administration. METHODS: We performed an open, 6 month, prospective evaluation of heme iron in HD patients who had been on maintenance IV iron therapy. IV iron was discontinued and replaced with oral heme iron. Serum iron level, hematocrit (Hct), and erythropoietin and IV iron dose were monitored. RESULTS: During 6 months, 4 of 37 patients (11%) dropped out because of insufficient iron supplementation or intolerance and 5 patients (14%) were dropped because of unrelated complications or protocol violation. A slight reduction in average transferrin saturation (TSAT) was seen early, but reversed, and no significant changes were seen in TSAT or Hct. A significant reduction in average serum ferritin level was seen at months 4 through 6 (P < 0.01). CONCLUSION: During the 6-month study period, heme iron polypeptide successfully replaced IV iron therapy in a majority of HD patients and maintained target Hcts with no concomitant use of IV iron. This treatment was associated with a significant increase in rHuEPO efficiency (P = 0.04). PMID- 12900817 TI - Levofloxacin pharmacokinetics in ESRD and removal by the cellulose acetate high performance-210 hemodialyzer. AB - BACKGROUND: No published data are available describing the pharmacokinetics of intravenous levofloxacin in patients with end-stage renal disease (ESRD). Objectives of this study are to determine the pharmacokinetics and dialytic clearance of levofloxacin and develop dosing strategies in these patients. METHODS: Eight noninfected subjects receiving long-term thrice-weekly hemodialysis, with no measurable residual renal function, were administered intravenous levofloxacin, 250 mg, over 1 hour after a scheduled hemodialysis session. Blood samples were collected serially during the interdialytic period, during the next intradialytic period, and immediately after the next hemodialysis session. Serum concentrations of levofloxacin were determined by high-performance liquid chromatography. Differential equations describing a 2-compartment open infusion pharmacokinetic model were fit to each individual subject's serum concentration-time data by iterative nonlinear weighted least-squares regression analysis using Adapt II (Biomedical Simulations Resource, University of Southern California, Los Angeles, CA). Ratios of maximum serum concentration (C(max)) to minimum inhibitory concentration (MIC) were calculated for common respiratory pathogens by using MIC for 90% of isolates (MIC90) data from published studies. RESULTS: All subjects completed the study, and no adverse events were reported. Median systemic clearance, volume of distribution at steady state, elimination half-life, and C(max) were 37.0 mL/min (range, 12.8 to 42.7 mL/min), 103.3 L (range, 39.8 to 139.3 L), 34.4 hours (range, 28.4 to 39.3 hours), and 5.2 microg/mL (range, 4.1 to 11.3 microg/mL), respectively. Median dialytic clearance and levofloxacin reduction ratios were 84.4 mL/min (range, 61.8 to 107.6 mL/min) and 0.244 (range, 0.181 to 0.412), respectively. Median C(max)-MIC90 ratios were 10 or greater for Haemophilus influenzae, Moraxella catarrhalis, Enterobacter cloacae, and Klebsiella pneumoniae, approximately 5 for Streptococcus pneumoniae, and less than 1 for Pseudomonas aeruginosa. CONCLUSION: The administration of levofloxacin to patients with ESRD as 500 mg initially, followed by 250 mg every 48 hours, will provide adequate C(max)-MIC ratios after the first and subsequent doses for most patients with respiratory tract infections caused by organisms with levofloxacin MICs of 1 microg/mL or less. PMID- 12900818 TI - Depression and its association with peritonitis in long-term peritoneal dialysis patients. AB - BACKGROUND: Depression is the most common psychological disorder among patients with end-stage renal disease and has been associated with mortality in patients maintained on hemodialysis therapy. Peritonitis is the leading cause of technique failure among long-term peritoneal dialysis (PD) patients. This prospective study is designed to examine the relationship between depression and peritonitis. METHODS: All patients on long-term PD therapy in our unit between January 1, 1997, and January 31, 2002, completed a Beck Depression Inventory (BDI) assessment at 6-month intervals. BDI scores were analyzed 2 ways. First, patients were placed into either group I (BDI score < 10) or group II (BDI score > or = 11) and were reclassified based on subsequent scores. Second, multivariable analysis was performed looking at initial BDI score as a risk factor for peritonitis, adjusting for age older than 65 years, diabetes, coronary artery disease, and race. RESULTS: One hundred sixty-two patients were enrolled, and 281 individual BDI assessments were completed. There was a significantly greater incidence of diabetes and coronary artery disease in group II. Rates for overall and gram-positive peritonitis were significantly greater in group II patients compared with group I patients. Using Cox regression, only BDI score of 11 or greater was associated with the development of peritonitis (hazard ratio, 2.7; 95% confidence interval, 1.2 to 6.0). CONCLUSION: There is an association between BDI score of 11 or greater and the development of peritonitis. Whether treatment of depression can impact on the rate of peritonitis remains to be examined. PMID- 12900819 TI - C-reactive protein, lipoprotein(a), homocysteine, and male sex contribute to carotid atherosclerosis in peritoneal dialysis patients. AB - BACKGROUND: In patients with end-stage renal disease, the morbidity and mortality of cardiovascular disease are substantially greater than in the general population. Advancement in understanding the pathogenesis of atherosclerotic vascular disease suggests a central role of inflammation in atherogenesis. However, clinical data evaluating the role of inflammation in atherogenesis are sparse in peritoneal dialysis (PD) patients. METHODS: We measured serum C reactive protein (CRP), intact parathyroid hormone, lipoprotein(a) [Lp(a)], interleukin-1 receptor antagonist (IL-1Ra), tumor necrosis factor soluble receptor (TNF-sR), fibrinogen, and plasma homocysteine (Hcy), as well as intima media thickness (IMT) and number of atherosclerotic plaques (plaque score [PS]) in the carotid arteries by means of carotid B-mode ultrasonography in 59 PD patients (35 men, 24 women; mean age, 52.4 years; average dialysis period, 36 months). All patients had chronic glomerulonephritis. RESULTS: Sixty-eight percent of PD patients had at least 1 plaque. Serum CRP level was greater than the upper limit of the normal range in 52.5% of patients. Compared with PD patients with normal CRP levels, concentrations of such proinflammatory cytokines as IL-1Ra and TNF-sR, Lp(a), and Hcy were increased in PD patients with elevated CRP levels. However, no differences in plasma fibrinogen and intact parathyroid hormone levels were found between PD patients with increased and normal CRP levels. In a multiple regression model, age, male sex, CRP level, and Lp(a) level were independent predictors of IMT. Similarly, male sex, CRP level, Lp(a) level, and Hcy level were independent correlates of PS. CONCLUSION: This study suggests that Lp(a) and Hcy levels and male sex, and especially CRP level, have an important role in carotid atherosclerosis in PD patients. PMID- 12900820 TI - Effects of exercise training on coronary heart disease risk factors in renal transplant recipients. AB - BACKGROUND: Multiple risk-factor interventions that include lifestyle changes have been proved to be effective in reducing risk profile in persons at high risk for developing coronary heart disease (CHD). There have not been similar studies involving transplant recipients. The purpose of this study is to examine effects of exercise training on cardiovascular risk profile during the first year after renal transplantation. We used traditional CHD risk factors and the Framingham CHD prediction methods. METHODS: Ninety-six transplant recipients were randomly assigned to 2 groups at 1 month posttransplantation: the exercise training (EX group; n = 51) and usual care groups (UC group; n = 45). Testing was performed at baseline and 12 months posttransplantation and included maximal exercise testing and evaluation of CHD risk factors and risk-factor categories, determined by means of the Framingham equations. RESULTS: Overall 10-year CHD risk score did not change in either group. All patients showed increases in total cholesterol (TC) level, high-density lipoprotein fraction of cholesterol (HDL-C), and body mass index over time. No differences were observed between groups, except for a trend toward greater improvement in HDL cholesterol levels in the EX group (P = 0.07). Significantly more patients in the EX group moved out of the high-risk category in TC-HDL ratio. All patients remained in the high-risk category for physical fitness. There was a significant negative correlation of CHD risk and maximal exercise capacity (r = -0.406; P < 0.001). CONCLUSION: Exercise training alone does not reduce CHD risk during the first year after transplantation. Research to determine the effects of multiple risk interventions during a longer period in transplant recipients is warranted. PMID- 12900821 TI - Long-term outcome of live donor kidney transplantation for renal amyloidosis. AB - BACKGROUND: The short-term outcome of kidney transplantation in patients with amyloidosis has been reported. The aim of this study is to investigate long-term results in patients with renal amyloidosis. METHODS: We studied results of renal transplantation in 23 amyloidotic transplant recipients compared with those in a control group of 47 nonamyloidotic patients. Amyloidosis was secondary to familial Mediterranean fever (FMF) in 16 patients, whereas it was primary (idiopathic) in 7 transplant recipients. The 2 groups were homogeneous regarding age, sex, HLA matching, immunosuppression, and duration of transplantation. RESULTS: Five- and 10-year actuarial graft survival rates were similar in both groups (79.35% versus 84.04% and 65.92% versus 56.61%, respectively ). Five- and 10-year actuarial patient survival rates also were similar (80% versus 94% and 68% versus 87%, respectively). Moreover, 72.4% of controls experienced at least 1 rejection episode, whereas only 43.5% of amyloidotic transplant recipients experienced 1 or more such events (P = 0.02). Nonetheless, mean serum creatinine concentrations did not differ between the 2 groups during the observation period. Maintenance colchicine therapy prevented the recurrence of both FMF symptoms and amyloidosis. Recurrence was documented in only 1 amyloidotic transplant recipient (4.3%) 10 years posttransplantation. Significant gastrointestinal (GI) problems were more frequent in amyloidotic patients (65% versus 38%; P = 0.03). Amyloidotic patients with GI problems, except for 2 patients, were administered cyclosporine. Eleven of these patients had FMF, which appeared to reflect the effects of both cyclosporine and colchicine. Infections were similar in the groups; whereas amyloidotic patients had significantly lower blood pressures. CONCLUSION: In our experience, long-term (5 to 10 years) outcome of live related donor kidney transplantation in patients with amyloidosis is similar to that in the general transplant population. PMID- 12900822 TI - Doxycycline decreases proteinuria in glomerulonephritis. AB - Treatment options for crescentic glomerulonephritis include the use of steroids, cytotoxic therapy, and, in severe cases, intravenous immunoglobulins and plasmapheresis. Injury and lysis of capillary glomerular basement membrane, which is made up of type IV collagen, laminin, fibronectin, and proteoglycans, by serine proteinases and matrix metalloproteinases (MMPs) likely is an important participant in the pathogenesis of crescentic glomerulonephritis. Tetracycline derivatives inhibit not only the activity of MMPs, but also their production, and have been investigated for the treatment of disorders in which the MMP system becomes amplified, such as degenerative osteoarthritis, periodontitis, cancer, and abdominal aortic aneurysm. We report an interesting case of crescentic glomerulonephritis in a young man who was treated with cyclophosphamide and prednisone. The patient developed steroid-induced acne that was treated with long term oral doxycycline therapy. During the period the patient was administered doxycycline, proteinuria decreased by 70% and recurred when doxycycline was stopped. To our knowledge, this is the first report of possible benefits of a metalloproteinase inhibitor (doxycycline) in glomerulonephritis in humans. Future studies are urgently required to explore the option of metalloproteinase inhibitors in the treatment of proliferative glomerulonephritis. PMID- 12900823 TI - Multicentric papillary renal carcinoma in renal allograft. AB - A renal transplant recipient with 13 years of excellent allograft function was found incidentally to have a malignant mass in his transplanted kidney. After resection, pathological analysis showed 29 separate lesions of renal cell carcinoma. All tumors were confined within the renal capsule. The majority of tumors (21 of 29 tumors) were chromophil basophilic carcinoma with papillary architecture, 5 tumors were clear cell, 2 tumors were mixed cell type, and 1 tumor was chromophil eosinophilic papillary carcinoma. These histological findings are similar to those reported in hereditary papillary renal carcinoma. To our knowledge, this is the first case of multicentric papillary renal carcinoma occurring in the renal allograft. We speculate that the allograft in this case is predisposed to malignant changes because of preexisting genetic mutations, as well as prolonged immunosuppression. PMID- 12900824 TI - Analgesics and renal disease in the postphenacetin era. PMID- 12900825 TI - Interventional nephrology: work in progress. PMID- 12900826 TI - Detecting early chronic kidney disease: are clinical laboratories a problem? PMID- 12900827 TI - An outline of essential topics in glomerular pathophysiology, diagnosis, and treatment for nephrology trainees. PMID- 12900828 TI - A dual pattern of immunofluorescence positivity. PMID- 12900830 TI - Venous access: are women equal? PMID- 12900832 TI - Say "no" and comorbidity. PMID- 12900834 TI - Monoclonal cryoglobulinemia in hepatitis C virus-associated, membranoproliferative glomerulonephritis. PMID- 12900841 TI - Quiz page. Acute interstitial nephritis and minimal change disease lesion, caused by NSAID injury. PMID- 12900843 TI - Visual impairment caused by retinal abnormalities in mesangiocapillary (membranoproliferative) glomerulonephritis type II ("dense deposit disease"). AB - Patients with mesangiocapillary glomerulonephritis (MCGN) type II usually present by early adulthood with hematuria, proteinuria, and renal impairment, and these features often are accompanied by a partial lipodystrophy and an autoantibody for the alternative complement pathway convertase (C3NeF). The diagnosis of MCGN type II depends on the demonstration of "dense deposits" in the glomerular basement membrane (GBM). Most patients also have multiple subretinal white spots or drusen that are histopathologically identical with the GBM deposits and evident ophthalmoscopically by the time renal failure develops. Initially visual acuity and visual fields are preserved, but fluorescein angiography and specialized tests of retinal function, such as dark adaptation, electroretinography, and electrooculography, may be abnormal and will worsen progressively. Over the next 20 years, vision often deteriorates because of retinal atrophy, and sometimes because of subretinal neovascular membranes, macular detachment, and central serous retinopathy. The authors describe a patient with MCGN type II who presented with renal failure and impaired vision at the age of 59. He already had widespread retinal atrophy, and subsequently a subretinal membrane developed. The drusen seen in MCGN type II, like the partial lipodystrophy, are a helpful clinical pointer to the diagnosis of this condition. All patients with MCGN type II should be warned of the risk of retinal complications and reviewed by an ophthalmologist at presentation and regularly after about 10 years to minimize the loss of visual acuity from complications of the retinopathy. PMID- 12900844 TI - Multiorgan failure during a sickle cell crisis in sickle/beta-thalassemia. AB - In contrast to the chronic nephropathy associated with sickle cell syndromes, acute renal failure and multiorgan dysfunction caused by acute sickling crisis are encountered infrequently. The authors present the first case of extensive multiorgan failure during a sickling episode in a patient with sickle/beta+thalassemia. The authors also review the interaction of the thalassemias with sickle cell disease and outline the distinctive course of their patient in comparison with previous reports. PMID- 12900846 TI - ARF after retrograde pyelography: a case report and literature review. AB - Acute renal failure (ARF) occasionally occurs after intravenous injection of contrast medium, but complications are rare after retrograde pyelography. After reviewing the reports in the English-language literature, the authors found very few on those complications after retrograde pyelography. The authors present a patient who had ARF after the technique. The patient had a history of hypopharyngeal cancer with underlying serum creatinine level at the high end of the normal limits. Bilateral flank pain and decreased urine amount were noted soon after the procedure of retrograde pyelography. Subsequently, blood urea nitrogen and creatinine levels both elevated, and hemodialysis was needed. Several days later, diuretic phase took place. Thereafter, the symptoms subsided gradually. Pyelorenal extravasation of contrast medium was remarkable during the procedure. There was no evidence of hydronephrosis during the course of ARF. Early awareness and management may prevent the complications of ARF such as acute lung edema and hyperkalemia. Therefore, clinical physicians should be aware of the occurrence of ARF and its clinical presentation after performing retrograde pyelography. PMID- 12900845 TI - Hyperkalemia with concomitant watery diarrhea: an unusual association. AB - Four patients presented to the emergency room with life-threatening hyperkalemia and concomitant watery diarrhea. Hypovolemia, acidosis, and renal insufficiency were present in all 4 cases. In 2 patients, hyperkalemia followed initiation of angiotensin-converting enzyme (ACE) inhibitor therapy, whereas 1 patient experienced hyperkalemia after a dose increase of an ACE inhibitor, and the fourth patient was on continuous ACE-inhibitor therapy at the time of the hyperkalemia episode. Two of the 3 patients with functioning kidneys required hemodialysis to correct the hyperkalemia, whereas the other patient was on long term hemodialysis therapy. In the 2 patients in whom transtubular potassium (K+) gradients were available, their values ranged far below normal, indicating tubular failure to secrete K+. This abnormality was attributed to decreased distal delivery of sodium and water and to renin/angiotensin II/aldosterone blockade. It has been proposed that aldosterone blockade impairs the capacity of the colonic epithelial cells to secrete K+. In all 4 patients the watery diarrhea ceased in parallel with the correction of serum K+ to normal values. It is suggested that hyperkalemia, most likely by stimulating intestinal motility, induced the watery diarrhea in all 4 patients. The watery diarrhea, however, failed to compensate for the renal tubular failure to secrete K+. PMID- 12900847 TI - Peritoneal-mediastinal leakage complication of peritoneal dialysis. AB - The authors report a case of mediastinal fluid collection resulting from peritoneal-mediastinal communication after continuous ambulatory peritoneal dialysis (CAPD). To the best of the authors' knowledge, this is the first reported case in the medical literature. A dry cough developed in the patient who had been receiving CAPD for 4 years. A mediastinal mass owing to peritoneal leakage of dialysate to the mediastinum was confirmed by a computed tomography scan taken 4 hours after the intraperitoneal infusion of contrast-mixed dialysate. The leakage persisted for 12 weeks after the discontinuation of CAPD fluid instillation. PMID- 12900849 TI - ANCA-positive crescentic glomerulonephritis associated with minocycline therapy. AB - Minocycline is an oral antibiotic widely used for the long-term treatment of acne and rheumatoid arthritis. A few patients develop antineutrophil cytoplasmic antibodies (ANCAs) during minocycline therapy. In this report, the authors describe a case of severe pauci-immune crescentic and necrotizing glomerulonephritis associated with positive cytoplasmic ANCA (C-ANCA) titers and proteinase 3 (PR3) levels after minocycline therapy. Discontinuation of minocycline and initiation of immunosuppressive treatment resulted in improvement of renal function and decline in C-ANCA titers and PR3 levels. A high degree of suspicion, testing for ANCA titers, prompt discontinuation of the drug, and initiation of immunosuppressive treatment are crucial to the diagnosis and treatment of drug-induced ANCA-associated glomerulonephritis. PMID- 12900848 TI - Renal manifestations of a mutation in the uromodulin (Tamm Horsfall protein) gene. AB - BACKGROUND: Uromodulin (Tamm Horsfall glycoprotein) is the most abundant protein found in normal human urine. Its function has yet to be determined. Identifying mutations in the uromodulin gene may be helpful in understanding the function of uromodulin. There has been 1 report of 4 families suffering from mutations in the uromodulin gene, resulting in the autosomal dominant transmission of hypouricosuric hyperuricemia and chronic renal failure. This case report describes another family with similar clinical manifestations. METHODS: A family was identified with clinical characteristics of hypouricosuric hyperuricemia and renal failure occurring in a mother and daughter. Clinical characteristics were identified, and laboratory studies were obtained in the proband and the proband's daughter. A genetic analysis was performed to evaluate for mutations in the uromodulin gene. RESULTS: The proband suffered from hyperuricemia at an early age and progressive renal failure with end-stage renal disease developing at age 49 years. The proband's daughter suffered from hyperuricemia, a reduced fractional excretion of uric acid, and mild renal insufficiency. A g.2105G > A mutation in exon 4 of the uromodulin gene resulting in a substitution of tyrosine for cysteine was identified in both the proband and the proband's daughter. The clinical characteristics were similar to those of other patients suffering from uromodulin mutations and to those of patients suffering from medullary cystic kidney disease type 2 and familial juvenile hyperuricemic nephropathy. CONCLUSION: Uromodulin associated kidney disease results in hyperuricemia and renal failure. The specific uromodulin mutation found in this family is consistent with the hypothesis that mutations disrupt highly conserved cysteine residues in the uromodulin protein. Potential mechanisms for these pathologic changes are discussed. The authors would appreciate referral of other families for screening for mutations. PMID- 12900850 TI - Isolated adrenocorticotropic hormone deficiency presenting with hypercalcemia in a patient on long-term hemodialysis. AB - The authors report on a 44-year-old female hemodialysis (HD) patient who presented with hypercalcemia secondary to isolated adrenocorticotropic hormone (ACTH) deficiency. She had been suffering from nausea and abdominal pain caused by recurrent esophageal ulcer. Blood calcium (Ca) adjusted for serum albumin concentration was increased to 14.9 mg/dL (3.72 mmol/L) concurrently with fever and hypotension. Serum intact parathyroid hormone (PTH)-related peptide was not elevated, but serum intact PTH and 1,25-(OH)2 vitamin D3 were decreased to 31 pg/mL (ng/L) and 8.1 pg/mL (2.6 pmol/L), respectively. Endocrinologic examination found that plasma ACTH was reduced below 5.0 pg/mL (0.22 pmol/L). A single ACTH stimulation normally increased blood cortisol, whereas a single corticotropin releasing hormone injection failed to increase plasma ACTH and cortisol. Pituitary magnetic resonance imaging disclosed no enlargement of pituitary gland. Circulating bone formation and absorption markers were not elevated. Blood Ca was normalized shortly after pamidronate disodium administration without glucocorticoid supplementation. This case suggested that secondary adrenal insufficiency caused by isolated ACTH deficiency could be an occult cause of severe hypercalcemia in HD subjects. PMID- 12900851 TI - Pharmaceutical and biomedical applications of chiral capillary electrophoresis and capillary electrochromatography: an update. AB - Capillary electrophoresis is often considered an ideal method for the chiral analysis of compounds due to the high separation power of the technique and has therefore found widespread acceptance for the analysis of drugs and pharmaceuticals. In contrast, capillary electrochromatography is still more or less in an infancy state searching for its place among the analytical separation techniques although interesting applications have been published. The present review summarizes recent developments and applications of chiral pharmaceutical analysis by electromigration techniques published in 2002 and early 2003. PMID- 12900852 TI - Microchip electrophoresis for chiral separations. AB - Microchip electrophoresis (MCE) is a promising new technique for the separation of enantiomers. This recently introduced technique enables chiral separations to be performed in seconds on tiny micromachined devices. This review is intended to give a brief introduction into the principles of chiral separations with MCE with regard to methodology and instrumentation. Different approaches to realize chiral separations in microfluidic devices are described and discussed. This review gives an overview of original work done in this field with emphasis on approaches to improve detection and resolution in chiral MCE. PMID- 12900853 TI - Chiral electromigration methods in food analysis. AB - This review article addresses the different chiral capillary electrophoretic methods that are being used for the study and characterization of foods and food compounds (e.g., amino acids, organic acids, sugars, pesticides). An updated overview, including works published till December 2002, on the principal applications of enantioselective procedures together with their main advantages and drawbacks in food analysis is provided. Some anticipated applications of chiral electromigration methods in food characterization are also discussed. PMID- 12900854 TI - Separation of basic drug enantiomers by capillary electrophoresis using chicken alpha1-acid glycoprotein: insight into chiral recognition mechanism. AB - Recombinant chicken alpha(1)-acid glycoprotein (alpha(1)-AGP) was prepared by the Escherichia coli expression system and completely deglycosylated alpha(1)-AGP (cd alpha(1)-AGP) was obtained by treatments of native alpha(1)-AGP with a mixture of endoglycosidase and N-glycosidase. The average molecular masses of chicken alpha(1)-AGP, cd-alpha(1)-AGP and recombinant alpha(1)-AGP were estimated to be about 29 200, 21 700 and 20 700, respectively, by matrix-assisted laser desorption-time of flight-mass spectrometry. We compared the chiral recognition ability of chicken alpha(1)-AGP, cd-alpha(1)-AGP and recombinant alpha(1)-AGP using them as chiral selectors in capillary electrophoresis. The chicken alpha(1) AGP showed higher resolution for eperisone, pindolol and tolperisone than cd alpha(1)-AGP or recombinant alpha(1)-AGP. Recombinant alpha(1)-AGP still showed chiral recognition for three basic drugs tested. By addition of propranolol as a competitor in the separation solution in CE, no enantioseparations of three basic drugs were observed with chicken alpha(1)-AGP, cd-alpha(1)-AGP or recombinant alpha(1)-AGP. These results reveal that the protein domain of the chicken alpha(1)-AGP is responsible for the chiral recognition ability, and that the chiral recognition site(s) for basic drugs exists on the protein domain. PMID- 12900856 TI - pH-dependent cyclodextrin capillary electrophoresis resolution of atropisomers. AB - Five noncommercial and four commercially available cyclodextrin (CD) derivatives were tested as chiral auxiliaries for the capillary electrophoretic (CE) resolution of racemic 1,1'-bi-(2-naphthol) (BN), 1,1'-binaphthyl-2,2'-diyl hydrogenphosphate (BNHP), and 1,1'-binaphthyl-2,2'-diamine (BNA) at pH 4.0, 6.5, and 8.6. The noncommercial CDs were ethyloxycarbonyl-gamma-CD (ethylcarbonate gamma-CD), dimethylamino ethyloxycarbonyl-beta-CD, a mercaptosuccinic acid derivative of beta-CD, a maleic acid derivative of beta-CD and heptakis(6-deoxy-6 amino)-beta-CD derivative with one amino group on the C-6 carbon of each glucose unit. Except for the latter, the remaining derivatives were synthesized for this work. Also commercially available methyl-beta-CD, hydroxypropyl-beta-CD and the native beta- and gamma-CDs were examined. Among the nine CDs tested, the maleic acid derivative of beta-CD gave the most interesting performances, since it resolved the atropisomers of BNA and BNHP in the same electrophoretic run at pH 4.0. It resolved the BNA racemate also at pH 6.5. Both the negatively charged CD tested were found to resolve anionic BNHP enantiomers, while positively charged CDs did not with cationic BNA. Several of the CDs investigated in this work were found to resolve the BNHP racemate, although at nonoptimal concentration. None of the experimented CDs was found to resolve the electrically neutral BN atropisomers pair at the three pHs considered, while some among these nine, experimented in a previous work, did so at higher pH. PMID- 12900855 TI - Mechanistic study of the enantiomeric recognition of a basic compound with negatively charged single-isomer gamma-cyclodextrin derivatives using capillary electrophoresis, nuclear magnetic resonance spectroscopy, and infrared spectroscopy. AB - The possible mechanisms for the chiral recognition of 2-(R)-N-[1-(6-aminopyridin 2-ylmethyl)piperidin-4-yl]-2-[(1R)-3,3-difluorocyclopentyl]-2-hydroxy-2 phenylacetamide (RR-M3), and its enantiomer (SS-M3) with octakis(2,3-di-O-acetyl 6-sulfo)-gamma-cyclodextrin (ODAS-gamma-CD) and octakis(6-sulfo)-gamma cycopdextrom enantiomer; (OS-gamma-CD), were investigated using capillary electrophoresis (CE), proton ((1)H), fluorine ((19)F) and carbon ((13)C) nuclear magnetic resonance spectroscopy (NMR), and infrared (IR) spectroscopy. Clear evidence for the formation of diastereomeric complexes between the enantiomers and the two CDs was observed. NMR spectra suggest that the phenyl and difluorocyclopentyl rings are involved in the complexation. The phenyl ring on the guest molecule is deeply penetrated into the cavity of OS-gamma-CD, but it is not included into the cavity of ODAS-gamma-CD. The continuous variation plots built based on the (1)H NMR and IR spectra indicate a 1:1 complex stoichiometric ratio of the M3 enantiomers for both CDs. The affinity of the enantiomers for the two CDs is opposite. PMID- 12900857 TI - Quantitative chiral analysis in capillary electrophoresis from unresolved peaks using derivative electropherograms, experimental design, and artificial neural networks. AB - Quantitative capillary electrophoretic analysis of chiral compounds might be difficult or even impossible when baseline separation is not reached. In this work, the use of n-th derivative of the electropherogram was studied and examined on model and experimental data. The electropherograms should be first smoothed using Savitzky-Golay method and the quantitative analysis is then possible using either a graphical method or multivariate calibration applying a combination of experimental design (ED) and artificial neural networks (ANNs). The best results were obtained for the first derivative, higher derivatives are not suitable because of noise accumulation. The method was applied to real experimental data to quantify chiral amino acids from unresolved peaks, but it is applicable for quantitative analysis of any other chiral analytes from poorly resolved peaks. Precision of analysis from partially resolved peaks reached was about +/- 3.2% relative standard deviation. PMID- 12900858 TI - Influence of peak measurement parameters on the quality of chiral electrophoretic separations. AB - The effect of nine peak measurement/analysis parameters on chiral capillary electrophoresis (CE), followed by UV detection, was studied. The parameters pertained UV-detection (detection wavelength, reference wavelength, and wavelength bandwidths), signal processing (data acquisition rate, type and amount of filtering) and peak detection (detection threshold and peak width). The influence of these factors on the chiral separation of dimethindene enantiomers was studied at two different concentrations (i.e., at high and low signal-to noise (S/N) ratio) by the means of experimental designs. The electropherogram characteristics considered were the resolution between the two enantiomers, the peak areas, and the S/N ratio. A D-optimal design was first used as screening design to identify the most critical parameters. Afterwards, a modelling of the different responses as a function of these critical parameters was performed based on the results of a face-centered central composite design. The results showed that the signal-processing parameters should be carefully selected when developing a CE separation since very important variations in the separation, the S/N ratio and the peak area of the substances can occur by setting these parameters at different levels. The detection wavelength should also be carefully chosen for optimal peak area measurement. The role of these parameters becomes more important with decreasing concentration of the analytes (i.e., low S/N ratio). This study showed that the peak measurement/analysis parameters should be optimized as the chemical and physical parameters of a method. They also should always be well specified in order to allow a good transfer of a method from one instrument to another. PMID- 12900859 TI - Coated microfluidic devices for improved chiral separations in microchip electrophoresis. AB - Chiral separations of fluorescein isothiocyanate-labeled amines have been performed in poly(vinyl alcohol) (PVA)-coated microfluidic glass chips. Baseline separation of enantiomers could be realized in coated devices while they could not be resolved in uncoated chips. The electroosmotic flow (EOF) in PVA-coated channels is suppressed over a wide pH range which leads to a considerable improved reproducibility of migration times in repetitive analysis. Due to the high resolution obtained in such devices, it was possible to reliable determine the enantiomeric purity with high accuracy. One percent of the minor enantiomer could be determined in the presence of large excess of the other enantiomer. As the EOF was suppressed, the anionic compounds were detected at the anode whereas the dominant EOF in uncoated devices resulted in an effective mobility to the cathode. Applying PVA-coated channels considerable improved precision of migration times was found. The relative standard deviation of migration times was below 1% in PVA-coated devices. Accordingly, excessive rinsing or etching steps in order to stabilize the EOF could be omitted while this was necessary for a reliable operation of uncoated devices. PMID- 12900860 TI - Synthesis, analytical characterization and initial capillary electrophoretic use in acidic background electrolytes of a new, single-isomer chiral resolving agent: hexakis(2,3-di-O-acetyl-6-O-sulfo)-alpha-cyclodextrin. AB - The sodium salt of hexakis(2,3-di-O-acetyl-6-O-sulfo)-alpha-CD (HxDAS), the first member of the family of single-isomer, fully sulfated alpha-CDs, has been synthesized and used for the initial capillary electrophoretic separation of the enantiomers of nonionic, weak acid, weak base, and ampholytic analytes. HxDAS complexes less strongly with many of the analytes tested than the analogous beta- and gamma-cyclodextrin derivatives, namely, heptakis(2,3-di-O-acetyl-6-O-sulfo) beta-cyclodextrin (HDAS) and octakis(2,3-di-O-acetyl-6-O-sulfo)-gamma cyclodextrin (ODAS). Nevertheless, it facilitated the separation of the enantiomers of a large number of weak electrolyte and nonelectrolyte analytes in acidic aqueous background electrolytes. For all analytes, the effective mobilities and separation selectivities as a function of the background electrolyte concentration of HxDAS followed the trends that were found for HDAS and ODAS. PMID- 12900861 TI - Enantioselectivity of alcohol-modified polymeric surfactants in micellar electrokinetic chromatography. AB - A novel method of modifying sodium undecanoyl-L-leucinate (SUL) micelles employed in chiral separation of analytes in micellar electrokinetic chromatography (MEKC) to enhance selectivity toward specific analytes is discussed. The current study aimed at modifying the SUL micelles by introducing different alcohols into the mono-SUL micelles. The micellar solutions were then polymerized in the presence of alcohols followed by postpolymerization extraction of the alcohols to yield alcohol-free polymeric surfactants (poly-L-SUL). The effects of hexanol (C(6)OH) and undecylenyl alcohol (C(11)OH) on micellar properties of this surfactant were investigated by use of surface tensiometry, fluorescence spectroscopy, pulsed field gradient-nuclear magnetic resonance (PFG-NMR), and MEKC. The surface tension and PFG-NMR studies indicated an increase in the critical micelle concentration (cmc) and micellar size upon increasing the alcohol concentration. Fluorescence measurements suggested that alcohols induce closely packed micellar structures. Coumarinic and benzoin derivatives, as well as (+/-)-1, 1'-binaphthyl 2,2'-dihydrogen phosphate (BNP) were used as test analytes for MEKC experiments. Examination of MEKC data showed remarkable resolutions and capacity factors of coumarinic derivatives obtained with modified poly-L-SUL as compared to the unmodified poly-L-SUL. Evaluation of fluorescence, PFG-NMR, and MEKC data suggest a strong correlation between the polarity and hydrodynamic radii of alcohol modified micelles and the resolution of the test analytes. PMID- 12900862 TI - Use of chiral zwitterionic surfactants for enantiomeric resolutions by capillary electrophoresis. AB - The enantiomeric resolution of 1,1'-binaphthyl-2,2'-diamine and Troger's base was investigated using the commercially available zwitterionic surfactants 3-[(3 cholamidopropyl)dimethylammonio]-1-propanesulphonate (CHAPS) and 3-[(3 cholamidopropyl)dimethylammonio]-2-hydroxy-1-propanesulphonate (CHAPSO). Resolution of the weakly basic chiral probes was achieved using varying concentrations of surfactant, above their critical micellar concentrations, in a phosphate buffer (pH 2.5; 100 mM) to ensure ionisation of the analytes. Both CHAPS and CHAPSO were employed in the absence of additional coselectors or surfactants as sole micellar-forming agents. The addition of organic modifiers, methanol and acetonitrile (ACN), to the background electrolyte (BGE) was found to have a detrimental effect on enantioselectivity presumably by alteration of the phase polarity. PMID- 12900863 TI - Polymeric alkenoxy amino acid surfactants: I. Highly selective class of molecular micelles for chiral separation of beta-blockers. AB - Two amino acid-based alkenoxy micelle polymers were synthesized for this study. These include polysodium N-undecenoxy carbonyl-L-leucinate (poly-L-SUCL) and polysodium N-undecenoxy carbonyl-L-isoleucinate (poly-L-SUCIL). The polymerization time and concentration of the synthesized micelle polymers were optimized by (1)H-nuclear magnetic resonance (NMR) and capillary electrophoresis (CE) experiments. Detailed physicochemical properties ((1)H NMR, critical micelle concentration (CMC), optical rotation, partial specific volume, aggregation number, and polarity) were determined, and these molecular micelles were introduced as a pseudostationary phase in micellar electrokinetic chromatography to study the molecular recognition and to develop a method for simultaneous separation of eight chiral beta-blockers. It is found that poly-L-SUCL gives overall better chiral resolution and wider chiral window than poly-L-SUCIL. After optimizing the type of micelle polymer, injection size and temperature, simultaneous separation and enantioseparation of eight beta-blockers were achieved in less than 35 min. A comparison with the amide-type surfactants of the same polar head group and alkyl chain length showed that carbamate-type surfactants always work better than the corresponding amide-type surfactant. PMID- 12900864 TI - Separation of some chiral flavanones by micellar electrokinetic chromatography. AB - Micellar electrokinetic chromatography (MEKC) was applied for enantioseparation of selected flavanones, including naringin, hesperidin, neohesperidin, naringenin, hesperetin, pinostrobin, isosakuranetin, eriodictyol, and homoeriodictyol. gamma-Cyclodextrin (gamma-CD) and sodium cholate (SCh) were used as chiral modifiers inducing enantioselectivity to the background electrolyte. From among many investigated selectors only these two appeared to possess the best enantioselective properties in respect to studied flavanones. The mechanisms of their action are a little different; SCh used above critical micelle point concentration forms chiral micelles itself while gamma-CD is deprived of this property and requires addition of surfactants as, e.g., sodium dodecyl sulfate. It was found that SCh enables separation of flavanone glycosides diastereomers while separation of enantiomers of flavanone aglycones may be achieved with gamma CD. Consideration of structural relation led to the suggestion that interaction of sugar moiety of glycosides with SCh micelles give rise to chiral recognition. MEKC appeared to be a suitable and efficient analytical tool to follow enantiomeric composition of flavanones. PMID- 12900865 TI - Enantiomeric impurity profiling in ephedrine samples by enantioselective capillary electrochromatography. AB - This study reports on the development and preliminary validation of a capillary electrochromatographic (CEC) method for the enantioselective impurity profiling of D-ephedrine. As chiral selector a novel low-molecular-weight strong chiral cation exchanger, based on penicillamine sulfonic acid, immobilized on thiol modified silica particles (3.5 microm) was employed. Under optimized conditions, the ephedrine enantiomers were separated on this chiral stationary phase (CSP) with an enantioselectivity of 1.11, an average efficiency of 321 550 plates per meter, and a resolution value of 4.77. A preliminary method validation was carried out to demonstrate the applicability of CEC for enantiomeric excess (ee) determination. Run-to-run repeatabilities (n = 5) reached relative standard deviation values (RSD) of 0.18 and 0.19% for the migration times of L- and D enantiomer, respectively, 0.3% for the resolution, and about 0.9% for the peak efficiencies. An approach called self-internal standard method was utilized to measure a standard calibration curve. Excellent linearity with a correlation coefficient of R(2) = 0.9998 was found for samples with concentrations in the range between 0.03 and 5 mg.mL(-1) D-ephedrine spiked with L-ephedrine at a constant concentration of 0.2 mg.mL(-1). The high loadability of the investigated CSP and good peak sensitivity allowed us to determine less than 0.1% enantiomeric impurity with good accuracy. The limit of detection (LOD) for the L-enantiomer in a 3 mg.mL(-1) D-ephedrine solution was found to be 0.035% (S/N = 3) and the limit of quantitation (LOQ) 0.058% (S/N = 5). For L-ephedrine samples the strong cation exchange (SCX)-type CSP with opposite configuration was utilized so that the enantiomeric impurity eluted before the main component peak yielding similar results in terms of separation and validation. Based on these results, the presented nonaqueous CEC methods are assessed as principally suitable for ee determination of ephedrine in terms of repeatability and method sensitivity. PMID- 12900866 TI - Chiral resolution of diastereomeric di- and tripeptides on a teicoplanin aglycone phase by capillary electrochromatography. AB - The chiral separation ability of a capillary packed with teicoplanin aglycone as a chiral stationary phase was investigated. This stationary phase was used successfully for chiral resolution of both diastereomeric dipeptides and tripeptides possessing one or two chiral centers. The composition of the mobile phase was shown to be crucial for separation. The use of reversed-phase mode was clearly superior to the polar-organic mode. The nature of the organic modifier was found to have a marked influence on separation. After optimizing conditions, all diasteromeric dipeptides and tripeptides investigated were baseline-resolved, however, it was not possible to find a uniform mobile phase showing optimal results for all peptides investigated. PMID- 12900867 TI - Cyclodextrin-modified monolithic columns for resolving dansyl amino acid enantiomers and positional isomers by capillary electrochromatography. AB - We describe beta- and gamma-cyclodextrins (beta- and gamma-CD)-modified monolithic columns prepared by sol-gel process and chemical modifications. The monolithic silica column was fabricated inside a fused-silica capillary with 100 microm inner diameter by sol-gel process. The monolithic silica matrix was chemically modified chiral selectors of beta- or gamma-CDs with a spacer of 3 glycidoxypropyltrimethoxysilane by on-column reactions. Gamma-CD-modified monolithic column has successfully been applied for the separation of dansyl amino acid enantiomers. Beta-CD-modified monolithic column has been used for the separation of the positional isomers of o-, m-, and p-cresols and the enantioseparation of racemates of benzoin and several dansyl amino acids by capillary electrochromatography, respectively. For the separation of neutral positional isomers, a positive electric field was applied. However, for the separation of negatively charged analytes, a negative electric field was applied at the inlet of column. The separation efficiency of 5.0 x 10(4) theoretical plates/m for dansyl-L-threonine was obtained at electric field strength of -300 V/cm in the mobile phase of 50 mM 2-(N-morpholino)ethanesulfonic acid (MES) Tris/methanol (70/30) buffer at pH 7.0. L-enantiomers were eluted as the first peak. Scanning electron micrograph showed that monolithic columns have the morphology of continuous skeleton and large through-pores. PMID- 12900868 TI - Capillary electrochromatographic separation of enantiomers on chemically bonded type of cellulose derivative chiral stationary phases with a positively charged spacer. AB - Positively charged chiral stationary phases (CSPs) were prepared for capillary electrochromatography (CEC) separation of enantiomers by chemically immobilizing cellulose derivatives onto diethylenetriaminopropylated silica (DEAPS) with tolylene-2,4-diisocyanate (TDI) as a spacer reagent. Anodic electroosmotic mobility was observed in both nonaqueous and aqueous mobile phases due to the positively charged amines on the surface of the prepared CSPs. For comparison, the traditionally used 3-aminopropyl silica (APS) was also adopted as the base material instead of DEAPS to prepare CSP. It was observed that the EOF on the DEAPS-based CSP was 18%-60% higher than that on the APS-based CSP under nonaqueous mobile phase conditions. Separation of enantiomers in CEC was performed on the positively charged CSPs with the nonaqueous mobile phases of pure ethanol or mixture of hexane-alcohol and the aqueous phases of acetonitrile water or 95% ethanol. Fast separation of enantiomers was achieved on the newly prepared CSPs. PMID- 12900869 TI - Enantioseparations of basic and bifunctional pharmaceuticals by capillary electrochromatography using polysaccharide stationary phases. AB - A fast screening strategy was developed in capillary electrochromatography (CEC) for the chiral separation of basic and bifunctional compounds. The screening conditions were determined on polysaccharide chiral stationary phases using 15 pharmaceutical compounds. The content and type of organic modifier, as well as the pH of the mobile phase appeared to have the largest influence on the chiral resolution. It was seen that for acidic compounds, our approach was not suitable. A generic mobile phase for basic and bifunctional compounds was determined. The testing on 20 additional compounds showed that the proposed mobile phase performed well since enantioselectivity was observed for 86% of the investigated compounds. A comparison of CEC and reversed-phase liquid chromatography (RPLC) results was attempted to demonstrate the potential of the used technique for chiral method development. PMID- 12900870 TI - Capillary electrophoresis to assess drug metabolism induced in vitro using single CYP450 enzymes (Supersomes): application to the chiral metabolism of mephenytoin and methadone. AB - Capillary electrophoresis (CE) with multiwavelength absorbance detection is demonstrated to be an effective tool for the assessment of in vitro drug metabolism studies using microsomes containing single human cytochrome P450 enzymes (CYPs) expressed in baculovirus-infected insect cells (Supersomes). Mephenytoin (MEPH), dextromethorphan, diclofenac, caffeine, and methadone (MET) were successfully applied as test substrates for CYP2C19, CYP2D6*1, CYP2C9*1, CYP1A2, and CYP3A4, respectively. For each system, the CE-based assay could be shown to permit the simultaneous analysis of the parent drug and its targeted metabolite. Using a chiral micellar electrokinetic capillary chromatography assay, the aromatic hydroxylation of MEPH catalyzed by CYP2C19 could thereby be confirmed to be highly stereoselective, an aspect that is in agreement with data obtained via urinary analysis after intake of racemic MEPH by extensive metabolizer phenotypes. The MET to 2-ethylidene-1,5-dimethyl-3,3 diphenylpyrrolidine (EDDP) conversion was investigated with a chiral zone electrophoresis assay. Incubation of racemic and nonracemic MET with CYP3A4 revealed no stereoselectivity for the transformation to EDDP, whereas no EDDP formation was observed with CYP1A2. CYP2C9 and CYP2C19 provided enhanced formation of R-EDDP and CYP2D6 incubation resulted in the preferential conversion to S-EDDP. Investigations using racemic MET and human liver microsomes revealed a modest stereoselectivity with an R/S EDDP ratio < 1 which is similar to the in vivo findings in urine. PMID- 12900871 TI - Capillary electrophoresis evidence for the stereoselective metabolism of itraconazole in man. AB - Itraconazole (ITC) is a hydrophobic antimycotic drug with three chiral centers that is used clinically as a stereoisomeric mixture. A chiral capillary electrophoretic method for the separation of ITC stereoisomers and those of its main metabolite hydroxyitraconazole (HITC) was developed to determine the stereoselective nature of the ITC to HITC biotransformation. The method is based on the formation of inclusion complexes of the target analytes with the negatively charged sulfated beta-cyclodextrin in the presence of moderate concentrations of methanol in a low-pH phosphate buffer. The addition of polyethylene glycol 4000 was found to be critical in obtaining baseline resolution of eight peaks, two from ITC, four from HITC, and two from R051012 (internal standard), in under 20 min. Application of the developed procedure to serum samples from patients being treated with ITC showed clearly the presence of a stereoselective component in the metabolism of this antimycotic drug. This could be shown from in vitro incubations with single enzyme Supersomes to be in part due to the stereoselective formation of HITC by the human CYP3A4 enzyme. For one patient, monitoring of the ITC and HITC concentrations and peak ratios over a 103 day period of treatment with ITC showed a strong dependency of the chiral ITC ratio to the concentration of ITC, while the dominant enantiomeric ratio of HITC was largely independent of the total HITC concentration. PMID- 12900872 TI - Assessment of the stereoselective metabolism of methaqualone in man by capillary electrophoresis. AB - Methaqualone (MQ) and its hydroxylated metabolites are quinazoline derivatives that exhibit atropisomerism. As a continuation of our previous work with these compounds (Electrophoresis 2001, 22, 3270-3280), chiral capillary zone electrophoresis with hydroxypropyl-beta-cyclodextrin as buffer additive and multiwavelength absorbance detection is shown to be an effective tool to provide insight into the stereoselectivity of the MQ metabolism. The five major monohydroxy MQ metabolites formed during biotransformation do not show enantiomerization at temperatures up to 85 degrees C. Enzymatic and acidic hydrolysis of urines that were collected after concomitant administration of 250 mg of MQ and 25 mg diphenhydramine (DH) chloride are both shown to provide stereoselective metabolic patterns with 4'-hydroxymethaqualone, the major urinary metabolite, being excreted almost exclusively as a single enantiomer. A stereoselectivity in the formation of 2'-hydroxymethaqualone and 2 hydroxymethaqualone was also observed in vitro using human liver microsomes and preparations containing the cytochrome P450 enzyme (CYP) CYP3A4 only. The presence of DH during incubation with human liver microsomes did not reveal a difference in the metabolic pattern obtained. Furthermore, CYP2D6 and CYP2C19 do not significantly contribute to the metabolism of MQ. This was independently observed in vitro and via analysis of urines of individuals that are either efficient metabolizer phenotypes or poor metabolizer phenotypes for the two polymorphic enzymes. Although interindividual differences in the monitored metabolic patterns were noted, no marked difference could be related to a CYP2D6 or CYP2C19 polymorphism. PMID- 12900873 TI - Enantiomeric separation of citalopram and its metabolites by capillary electrophoresis. AB - A simple and fast capillary electrophoretic method has been developed for the enantioselective separation of citalopram and its main metabolites, namely N desmethylcitalopram and N,N-didesmethylcitalopram, using beta-cyclodextrin (beta CD) sulfate as the chiral selector. For method optimisation several parameters were investigated, such as CD and buffer concentration, buffer pH, and capillary temperature. Baseline enantioseparation of the racemic compounds was achieved in less than 6 min using a fused-silica capillary, filled with a background electrolyte consisting of a 35 mM phosphate buffer at pH 2.5 supplemented with 1% w/v beta-CD sulfate and 0.05% w/v beta-CD at 25 degrees C and applying a voltage of -20 kV. A fast separation method for citalopram was also optimized and applied to the analysis of pharmaceutical formulations. Racemic citalopram was resolved in its enantiomers in less than 1.5 min using short-end injection (8.5 cm, effective length) running the experiments in a background electrolyte composed of a 25 mM citrate buffer at pH 5.5 and 0.04% w/v beta-CD sulfate at a temperature of 10 degrees C. PMID- 12900874 TI - Enantioseparation of warfarin and its metabolites by capillary zone electrophoresis. AB - A capillary zone electrophoresis (CZE) method with direct ultraviolet (UV) absorbance detection is presented for the simultaneous enantiomeric separation of warfarin and its main metabolites, including warfarin alcohols, 4'-, 6-, and 7 hydroxywarfarin, using highly sulfated beta-cyclodextrin (HS-beta-CD) as the chiral selector. This chiral separation method was optimized in terms of the electrophoretic parameters, which included the concentration of HS-beta-CD used, the type and composition of organic modifier added to the background electrolyte (BGE) buffer, and the BGE buffer pH. Chiral separation of warfarin and its major metabolites was achieved with high resolution, selectivity, efficiency, repeatability, and reproducibility. This optimized chiral analysis of warfarin along with its metabolites was completed within a satisfactory electrophoresis time of 20 min. PMID- 12900875 TI - Quantitation of talinolol and other beta-blockers by capillary electrophoresis for in vitro drug absorption studies. AB - A capillary zone electrophoresis method is described for the enantioseparation of talinolol using heptakis(2,3-diacetyl-6-sulfo)-beta-cyclodextrin (HDAS-beta-CD) as a chiral selector. After liquid-liquid extraction of talinolol from physiological solution, electrokinetic injection was employed to improve the sensitivity. The use of a coated capillary was necessary to achieve stable and reproducible enantioseparations. A baseline separation of the talinolol enantiomers was achieved in less than 10 min using 100 mM phosphate solution as background electrolyte and pH 3.5, at the presence of 3.0 mM HDAS-beta-CD and at 20 degrees C. In addition, this analytical condition proved to be useful for the enantioseparation of a number of other beta-blocking agents such as alprenolol, atenolol, bisoprolol, celiprolol, metipranolol, oxprenolol, and sotalol. For determining talinolol, the method could be validated in terms of precision, accuracy and linearity, and was found to be suitable in determination of talinolol enantiomers in highly diluted samples obtained from in vitro experiments. PMID- 12900876 TI - Infinite enantiomeric resolution of basic compounds using highly sulfated cyclodextrin as chiral selector in capillary electrophoresis. AB - Four chiral basic analytes, namely methadone, fluoxetine, venlafaxine, and tramadol, were selected as model compounds for investigating their stereoselective separation with highly sulfated gamma-cyclodextrin (HS gamma-CD) by capillary electrophoresis (CE)-UV and CE-mass spectrometry (MS). At high concentration of chiral selector, the preferentially bonded enantiomer migrated faster in the anodic mode to the detector and high resolutions were obtained for all analytes. In the cathodic mode, at lower highly sulphated cyclodextrin (HS CD) concentration, basic compounds could be detected, with the weakly bonded enantiomer migrating first (enantiomeric migration order inversion). It was also then possible, at intermediate HS-CD concentration, that only one enantiomer migrated to the detector as cation while the other enantiomer complexed with the CD was negatively charged and presented an opposite mobility. The latter never reached the detector achieving a perfect enantiomeric selectivity. Infinite chiral resolutions were thus achieved by CE-UV as well as by CE-electrospray ionisation (ESI)-MS where concentrations of HS-CD were adapted according to the negative contribution of the nebulization gas pressure of the interface. PMID- 12900877 TI - Simultaneous chiral discrimination of multiple profens by cyclodextrin-modified capillary electrophoresis in normal and reversed polarity modes. AB - Simultaneous enantioseparations of nine profens for their accurate chiral discrimination were achieved by capillary electrophoresis (CE) in the normal polarity (NP) mode with a single cyclodextrin (CD) system and in the reversed polarity (RP) mode with a dual CD system. The single CD system in the NP mode employed heptakis(2,3,6-tri-O-methyl)-beta-cyclodextrin (TMbetaCD) added at 75 mM 100 mM 2-(N-morpholino)ethanesulfonic acid buffer (pH 6.0) as the optimum run buffer. The dual CD system operated in the RP mode used 30 mM TMbetaCD and 1.0% anionic carboxymethyl-beta-cyclodextrin dissolved in pH 3.0, 100 mM phosphoric acid-triethanolamine buffer containing 0.01% hexadimethrine bromide added to reverse the electroosmotic flow. Fairly good enantiomeric resolutions and the opposite enantiomer migration orders were achieved in the two modes. Relative migration times to internal standard under respective optimum conditions were characteristic of each enantiomer with good precision (< 2% relative standard deviation, RSD), thereby enabling to crosscheck the chemical identification of profens and also their accurate chiralities. The method linearity in the two modes was found to be adequate (r > or = 0.9991) for the chiral assay of the profens investigated. Simultaneous enantiomeric purity test of ibuprofen, ketoprofen and flurbiprofen in a mixture was feasible in a single analysis by the present method. PMID- 12900878 TI - Enantioseparation of dihydrofurocoumarin derivatives by various separation modes of capillary electrophoresis. AB - Chiral dihydrofurocoumarin compounds are currently the focus of industrial and pharmacological research. These derivatives have been shown to possess many physiological properties that could be medically beneficial. This work proposes four different chiral separation methods using capillary electrophoresis and micellar capillary electrophoresis (MCE). Several different cyclodextrin chiral selectors were examined to evaluate their effectiveness in the enantioseparation of dihydrofurocoumarins. In addition, the effects of the chiral selector concentration, the presence of an organic modifier, run buffer pH, and in two cases, the ratio between the chiral selector and an additional charged pseudophase were investigated. Overall, the best separations for this class of chiral compounds were achieved using sulfated beta-cyclodextrins at low pH in the reversed polarity mode. PMID- 12900879 TI - Comparison of charged cyclodextrin derivatives for the chiral separation of atropisomeric polychlorinated biphenyls by capillary electrophoresis. AB - Charged cyclodextrin (CD) derivatives were used as chiral selectors in electrokinetic chromatography (EKC) for the chiral separation of highly hydrophobic neutral racemates such as atropisomeric polychlorinated biphenyls (PCBs). beta-CD-phosphated, beta-CD sulfated, succinylated-gamma-CD (Succ-gamma CD) and succinylated-beta-CD (Succ-beta-CD) were used as anionic CDs. As cationic CD, 6-monodeoxy-6-monoamino-beta-CD (beta-CD-NH(2)) was tested for the first time in order to separate PCBs. From the different CD derivatives employed, the best separations were obtained with the cationic CD derivative. Thus, the use of beta CD-NH(2 )in phosphate buffer at pH 2.0 containing urea allowed the chiral recognition of eleven PCBs (45, 84, 88, 91, 95, 131, 136, 144, 149, 176, and 197). In this case, the addition of 2 M urea to the buffer solution was crucial to achieve the chiral separation of PCBs. The addition of acetonitrile to 10 mM phosphate buffer (pH 2.0) with 30 mM beta-CD-NH(2) and 2 M urea improved considerably the chiral resolution obtained for PCBs 91, 95, 136, 144, 149, and 197 although an increase in the analysis time was also observed. All the results obtained were compared with those previously obtained with the dual CD system carboxymethyl-gamma-CD/beta-CD. PMID- 12900880 TI - Chiral characterization of deprenyl-N-oxide and other deprenyl metabolites by capillary electrophoresis using a dual cyclodextrin system in rat urine. AB - A chiral capillary electrophoresis method has been developed for the simultaneous separation of the enantiomers of deprenyl and eight of its metabolites, among them the recently described metabolite deprenyl-N-oxide. Although heptakis-(2,6 di-O-methyl)-beta-cyclodextrin (DIMEB) was suitable for the enantioresolution of deprenyl and its dealkylated derivatives, the enantiomers of deprenyl-N-oxide were just partly resolved. Carboxymethyl-beta-cyclodextrin (CMBCD) in as low as 2 mM concentration was capable of the enantiomer separation of all the nine examined compounds, however co-migration of 1R,2S-(-)-norephedrine and 1R,2R-(-) pseudoephedrine, as well as 1S,2R-(+)-ephedrine and R-(-)-amphetamine was observed. This problem could be overcome by the use of a dual cyclodextrin system containing 4 mM DIMEB in addition to 2 mM CMBCD; simultaneous separation of all the compounds could be achieved. The optimized method was used for the analysis of rat urine samples after 10 days of treatment of animals with either R-(-)- or S-(+)-deprenyl. The stereospecific biotransformation of both deprenyl enantiomers was confirmed, and the stereoselectivity of N-oxide formation was demonstrated. PMID- 12900881 TI - Fast enantiomeric separation with vancomycin as chiral additive by co electroosmotic flow capillary electrophoresis: increase of the detection sensitivity by the partial filling technique. AB - A fast and sensitive method is described by using vancomycin as a chiral additive for enantiomeric separation by capillary electrophoresis (CE). In order to overcome disadvantages associated with use of vancomycin as chiral additive in CE, several strategies including the dynamic coating technique, the co electroosmotic flow technique, and the partial filling technique were employed sequentially in this method. Using the polycationic polymer hexadimethrine bromide (HDB) as a buffer additive, the capillary wall was dynamically coated with a thin film formed by the adsorbed HDB. Consequently, the adsorption of vancomycin onto the capillary wall was minimized via electrostatic repulsion between the coating of the capillary wall and the vancomycin molecule. In addition, the reversed electroosmotic flow (from cathode to anode) produced by the positively charged capillary wall migrates in the same direction of negatively charged analytes (co-electroosmotic flow electrophoresis). Thereby the electrophoretic mobility of negatively charged analytes were drastically accelerated leading to a short separation time of less than 3.4 min. The separation time was further reduced by the use of a short-end-injection technique. For example, the analysis time was achieved by as short as 55 s for a baseline separation of dansyl-alpha-amino-n-butyric acid. Concurrently, the partial filling technique was used to avoid the loss of detection sensitivity caused by the presence of vancomycin in the running buffer. The effect of several parameters, such as HDB concentration, buffer pH, plug length of the chiral selector, concentration of the chiral selector and applied voltage, on enantioselectivity were investigated toward optimization. Besides the advantage of a very short separation time, the method is characterized by high detection sensitivity, high selectivity, and high efficiency. PMID- 12900882 TI - Rapid determination of salbutamol in pharmaceutical preparations by chiral capillary electrophoresis. AB - A fast and simple method of chiral capillary electrophoresis (CE) has been applied to the analysis of salbutamol in different pharmaceutical preparations. Using of a 25 mM acetate buffer (pH 5.0), containing 13.1 mg/mL carboxymethyl beta-cyclodextrin (CM-beta-CD), an applied voltage of 20 kV and a temperature of 25 degrees C, the enantiomers of salbutamol could be separated in about 2 min. Three different pharmaceutical preparations (two syrups, one oral solution, and two kind of tablets) containing a racemate of salbutamol were injected directly in the CE system, following dilution in dimethyl sulfoxide (DMSO). Appreciable differences in the retention times were observed for salbutamol enantiomers in the different formulations studied, which were attributed to the effect of the matrix components on the electrophoretic mobility. The standard addition method was used for the calibration due to the existence of matrix interferences. Finally, the stability of the enantiomers of salbutamol in the oral solution was studied calculating the enantiomeric ratio values when the solution was injected immediately after being opened in the first case and after being opened and stored in the fridge for two months in the second case. PMID- 12900883 TI - Determination of L-hyoscyamine in atropine and D-hyoscyamine in L-hyoscyamine by chiral capillary electrophoresis as an alternative to polarimetry. AB - A method for the separation of atropine enantiomers, D- and L-hyoscyamine by capillary electrophoresis (CE) has been developed and validated. The advantages of the CE method compared with polarimetry include smaller amounts of analytes and a lower limit of detection of the unwanted enantiomer. Moreover, the present method enables a baseline separation of the analytes and tropic acid, one of the typical impurities of atropine. The developed enantioseparation of atropine was performed using a commercially available sulfated beta-cyclodextrin and was validated for the determination of L-hyoscyamine in atropine as well as for the enantiomeric purity of L-hyoscyamine. PMID- 12900884 TI - Separation of aminoalkanephosphonic acid enantiomers by indirect UV detection capillary electrophoresis with application of cyclodextrins. AB - Indirect UV detection capillary electrophoresis (CE) was used for the separation of aminoalkanephosphonic acid (AP) enantiomers by applying commercially available cyclodextrins as chiral discriminators. The results show that the separation of the enantiomers depends on pH of the background electrolyte, the molar ratio of cyclodextrin to aminophosphonic acid, and on the type of the applied chiral selector. Optimization of process conditions allowed enantiomeric baseline separation or partial separation of 12 out of 14 alpha-aminophosphonic acids studied. This type of CE might therefore be successfully used for routine determination of enantiomeric purity of aminophosphonic acids. PMID- 12900885 TI - Enantiomeric separation of chiral peptide nucleic acid monomers by capillary electrophoresis with charged cyclodextrins. AB - Direct chiral separation of chiral peptide nucleic acid (PNA) monomers has been achieved for the first time by capillary electrophoresis (CE) with charged cyclodextrins as chiral selectors added to the electrophoretic buffer. Selectively modified 6-deoxy-6-N-histamino-beta-cyclodextrin and sulfobutyl ether beta-CD were successfully used as chiral selectors for the enantiomeric separation of chiral monomers based on different aminoethylamino acids bearing thymine or adenine as nucleobases. Chiral separations were obtained at low selector concentrations (1-3 mM) with good enantioselectivity and resolution factors. Separations were optimized as a function of pH in order to exploit the effect of the electrostatic interactions between the oppositely charged selector and selectand. The method has been applied to the analysis of the enantiomeric excess of chiral monomers used for the solid phase synthesis of chiral PNA oligomers. CE chiral analysis showed that a very high enantiomeric purity was generally achieved in the synthesis of all monomers, except for histidine and aspartic acid based monomers in which ca. 10% of the "wrong" enantiomer was always present. PMID- 12900887 TI - Direct chiral resolution of tartaric acid by ion-pair capillary electrophoresis using an aqueous background electrolyte with (1R,2R)-(-)-1,2-diaminocyclohexane as a chiral counterion. AB - Chiral resolution of native DL-tartaric acid was achieved by ion-pair capillary electrophoresis (CE) using an aqueous-ethanol background electrolyte with (1R,2R) (-)-1,2-diaminocyclohexane (R-DACH) as a chiral counterion. Factors affecting chiral resolution and migration time of tartaric acid were studied. By increasing the viscosity of the background electrolyte and the ion-pair formation, using organic solvents with a lower relative dielectric constant, resulted in a longer migration time. The optimum conditions for both high resolution and short migration time of tartaric acid were found to be a mixture of 65% v/v ethanol and 35% v/v aqueous solution containing 30 mM R-DACH and 75 mM phosphoric acid (pH 5.1) with an applied voltage of -30 kV at 25 degrees C, using direct detection at 200 nm. By using this system, the resolution (Rs) of racemic tartaric acid was approximately 1. The electrophoretic patterns of tartaric and malic acids suggest that two carboxyl groups and two hydroxyl groups of tartaric acid are associated with the enantioseparation of tartaric acid by the proposed CE method. PMID- 12900886 TI - Capillary electrophoresis as a probe of enantiospecific interactions between photoactive transition metal complexes and DNA. AB - Recently, we have demonstrated the capacity to separate chiral transition metal (TM) complexes of the type [M(diimine)(3)](n+) using CE buffers containing chiral tartrate salts. In separate work, several chromium(III)-tris-diimine complexes in particular have been shown to bind enantioselectively with calf-thymus (CT) DNA, and a qualitative assessment of the relative strength and enantiospecificity of this interaction is of significant interest in the characterization of these complexes as potential DNA photocleavage agents. Here, we describe two convenient approaches to investigate such binding behavior using chiral CE. For complexes with lower DNA affinities exhibiting primarily surface binding, DNA itself is used as the chiral resolving agent in the electrophoretic buffer. In this approach, resolution of the TM complexes into their Lambda and Delta isomers is achieved with the isomer eluting later exhibiting superior binding affinity toward DNA. For more strongly bound TM complexes containing ligands known to intercalate with DNA, the [Cr(diimine)(3)](3+) complexes are preincubated with oligonucleotide and subsequently enantiomerically resolved in a dibenzoyl-L tartrate buffer system that facilitates analysis of the unbound TM species only. Differences in isomer binding affinity are distinguished by the relative peak areas of the Lambda- and Delta-isomers, and relative binding strengths of different complexes can be inferred from comparison of the total amount of unbound complex at equivalent DNA/TM ratios. PMID- 12900888 TI - Chiral separation of diastereomeric flavanone-7-O-glycosides in citrus by capillary electrophoresis. AB - The 2S- and 2R-diastereomers of major flavanone-7-O-glycosides found in sweet orange (Citrus sinensis), mandarine (Citrus deliciosa), grapefruit (Citrus paradisi), lemon (Citrus limon), and sour or bitter orange juice (Citrus aurantium) were separated for the first time by chiral capillary electrophoresis (CE) employing various buffers with combined chiral selectors. Native cyclodextrins (CDs), neutral and charged CD derivatives were examined as chiral additives to the background electrolyte (BGE). Separation efficiency has not proved satisfactory with one single CD as chiral selector in the buffer, a full and simultaneous separation could often be achieved only by using combined buffer with two different CDs. Chiral separation of major flavanones in sweet orange, mandarine and grapefruit juices raised more difficulties than in lemon and sour orange juices as narirutin will not readily build complexes with most CDs. Diastereomeric flavanones of mature and immature grapefruits were compared and some differences were found: naringin showed different diastereomeric ratio and 2S-prunin appeared only in immature grapefruit. Marmalade was also examined by chiral CE. Its major flavanones corresponded to flavanone pattern of mixed sour and sweet oranges. PMID- 12900889 TI - Study on thioridazine 5-sulfoxide epimerization and degradation by capillary electrophoresis. AB - We present a method for the stereoselective analysis of thioridazine 5-sulfoxide (THD 5-SO) in human plasma based on liquid-liquid extraction with diethyl ether and chiral resolution of the stereoisomers by capillary electrophoresis using hydroxypropyl-beta-cyclodextrin and sulfated beta-cyclodextrin as chiral selectors. The method showed recovery rates of 85.5% for both THD 5-SO (slow eluting, SE) enantiomers. The coefficients of variation observed in the precision studies, as well as the accuracy values, were below 10%. After validation, the method was used to study the stability and configurational changes of this THD metabolite. Our results showed that both enantiomers of THD 5-SO (SE) were stable under conditions of variation of temperatures (38 degrees C, 4 degrees C and -20 degrees C), pH (5.0, 7.0 and 8.5) and ionic strengths (0.2, 0.5 and 1.0 mol/L). The influence of light on the stability of the THD 5-SO (SE) stereoisomers was also studied using standard solutions prepared in methanol and an inversion in configuration was observed under UV light (254 and 366 nm). PMID- 12900893 TI - A novel 5q35.3 subtelomeric deletion syndrome. AB - We observed a novel 3.5 Mb 5q subtelomeric deletion in a 3-year-old girl with developmental delay, hypotonia and multiple minor anomalies. Comparison of her phenotype with the few published patients with terminal 5q35 deletions revealed several overlapping features, but also showed remarkable differences such as shortness of stature versus macrosomia. After the report of 5q35.3 microdeletions in Sotos syndrome we integrated the published BACs into the public draft sequence and exactly mapped the deletion size in our patient by FISH analysis with 15 BAC probes. We demonstrated that the deletion in our patient is immediately adjacent to the reported Sotos syndrome deletion site. Subtracting the symptoms of Sotos syndrome from the published patients with larger 5q35.3 deletions allowed us to delineate a distinct phenotype of prenatal lymphedema with increased nuchal translucency, pronounced muscular hypotonia and delay of reaching motor milestones, but speech development within normal limits, wide fontanels, failure to thrive with postnatal short stature, and multiple minor anomalies such as mildly bell-shaped chest, minor congenital heart disease, and a distinct facial gestalt, associated with the novel 3.5 Mb cryptic deletion. We further showed in our patient that the deletion of the LCT(4) synthase gene results in a reduction of cysteinyl leukotriene synthesis to about 65% compared to normal values. The prenatal nuchal lymphedema associated with this deletion syndrome my be related to the deletion of the FLT4 gene causing autosomal dominant primary lymphedema and contributes to the differential diagnosis of increased fetal nuchal translucency. PMID- 12900894 TI - Clinical and molecular analysis of Grebe acromesomelic dysplasia in an Omani family. AB - Grebe syndrome is a rare autosomal recessive acromesomelic dysplasia. The syndrome was studied clinically, radiographically, and genetically in an Omani family with four affected children. The affected persons had normal axial skeletons, severely shortened, and deformed limbs with severity increasing in a proximo-distal gradient, and subluxated joints. The humeri and femora were hypoplastic with distal malformations. The radii/ulnae were shortened and deformed whereas carpal bones were invariably rudimentary or absent. The tibiae appeared rudimentary; fibulae were absent in two children, and some tarsal and metatarsal bones were absent. The proximal and middle phalanges were absent while the distal phalanges were present. The father and mother had short first metacarpal and middle phalynx of the fifth finger and hallux valgus respectively. Transition A1137G and deletion delG1144 mutations in the gene encoding the cartilage-derived morphogenetic protein-1 (CDMP-1) were identified in this family. The A1137G is a silent mutation coding for lysine, whereas the delG1144 predicts a frameshift mutation resulting in a presumable loss of the CDMP-1 biologically active carboxy-terminal domain. The affected siblings were homozygous for the delG1144 mutation while parents were heterozygous. PMID- 12900896 TI - Turner phenotype in a girl with a 45,X/46,XX/47,XX,+18 mosaicism. AB - We report a girl with Turner syndrome phenotype, whose karyotype on amniocyte culture was 45,X, while cytogenetic analysis on peripheral blood lymphocytes showed the presence of a mosaic chromosome constitution with three different cell lines: 45,X[5]/46,XX[3]/47,XX,+18 [35]. No signs of trisomy 18 were observed and a follow up during childhood revealed normal psychomotor development. Parental origin and mechanism of formation were studied using high polymorphic microsatellites and Quantitative Fluorescent PCR. The 18-trisomic cells showed one paternal allele and two maternal homozygous alleles at different loci of chromosome 18, suggesting a maternal M-II meiotic or a postzygotic error. A biparental origin of the X-alleles in the trisomic cells were determined, being the paternal allele retained in the 45,X cells. The possible mechanism of formation implying meiotic and/or mitotic errors is discussed. PMID- 12900895 TI - Linkage study between congenital cataracts and five crystallin loci. AB - Congenital cataracts are a common cause of preventable blindness in children. We studied autosomal dominant congenital cataracts in 38 families and examined linkage between cataract loci and the crystallin genes on chromosomes 2, 11, 17, 21, and 22. We used clinical information to group families with phenotypically similar cataracts and analyzed the genetic data in these groups. Although LOD scores > 3.0 were not obtained, we found some support for linkage to four of the chromosomal regions examined, namely 2q33-35, 17q11.2-12, 21q22.3, and 22q11.2. PMID- 12900897 TI - Colobomatous macrophthalmia with microcornea syndrome: report of a new pedigree. AB - Colobomatous macrophthalmia with microcornea syndrome (MIM 602499) is a very rare eye malformation. To date, only two families with a total number of eight patients have been reported. We report on 13 additional cases in a single three generation family and describe the eye findings and quantitative parameters of corneal curvature and axial ocular dimensions. All affected relatives had bilateral involvement with typical inferonasal iris coloboma, chorioretinal coloboma, microcornea, and varying degrees of axial enlargement associated with myopia. Additional findings included flatter corneal curvatures and shallower anterior chambers. Iridocorneal angle abnormalities associated with elevation of intraocular pressure were detected in three patients. The pedigree confirms the autosomal dominant pattern of inheritance with complete penetrance. PMID- 12900898 TI - Myoclonus in a patient with a deletion of the epsilon-sarcoglycan locus on chromosome 7q21. AB - Autosomal dominant myoclonus-dystonia syndrome (MDS) is characterized by myoclonic and/or dystonic movements with onset as early as infancy. In most families, MDS is caused by mutations in the gene SGCE, which encodes epsilon sarcoglycan and is located on chromosome 7q21. Data from several sources, including multi-generation pedigrees revealing parent-of-origin effects on MDS penetrance, suggest that SGCE is maternally imprinted. We present a 32-month-old patient with an interstitial deletion affecting chromosome 7q21, and a phenotype including myoclonus, microcephaly, short stature, dysmorphic face and language delay. We used fluorescence in situ hybridization (FISH) to estimate the size of our patient's deletion (9.0-15 Mbp) and to confirm absence of SGCE on the affected chromosome. Polymerase chain reaction (PCR) analysis of polymorphic markers in the region revealed that the paternally inherited chromosome contained the deletion, consistent with a model of maternal SGCE imprinting. Our patient is the first case of MDS caused by complete deletion of SGCE, and represents a new contiguous gene disorder. The case underscores the need to consider chromosomal deletions in patients whose phenotypes are more complex than the classic presentation of a known disease. PMID- 12900899 TI - Novel mutation in the gene encoding c-Abl-binding protein SH3BP2 causes cherubism. AB - Cherubism is a rare autosomal dominant inherited condition caused by mutations in the c-Abl-binding protein SH3BP2. It is characterized by multiple cystic giant cell lesions of the jaw appearing in early childhood with stabilization and remission after puberty. In the present study, we used direct sequence analysis of the SH3BP2 gene of several individuals from a family with cherubism to search for additional SH3BP2 mutations resulting in cherubism. In affected relatives, we found a previously unreported G to A transition in exon 9 leading to a Gly to Arg substitution at amino acid position 420. G420R has been reported previously with a G to C transversion. To date there have been no disease causing mutations outside exon 9. Therefore, the amino acid sequence from positions 415 to 420 may represent a specific protein domain which, when disrupted, leads to the cherubism phenotype. PMID- 12900900 TI - Beare-Stevenson syndrome: Two South American patients with FGFR2 analysis. AB - We report two patients with Beare-Stevenson syndrome. This syndrome presents craniosynostosis with or without clover-leaf skull, craniofacial anomalies, cutis gyrata, acanthosis nigricans, prominent umbilical stump, furrowed palms and soles, genital and anal anomalies. Both female newborn patients presented at birth with craniofacial anomalies, variable cutis gyrata in forehead and preauricular regions, prominent umbilical stump and anogenital anomalies. Furrowed palms and soles were also observed. The radiologic examination showed a cloverleaf-form craniosynostosis. Chromosomes were normal. They were born with respiratory distress and were connected to mechanical ventilation for ventilatory support. Both of them died in 50 days after birth due to secondary complications. The molecular analysis of these patients identified the mutation Tyr375Cys in the FGFR2 gene. PMID- 12900901 TI - Non-syndromic hemihyperplasia in a male and his mother. AB - We present two family members with non-syndromic hemihyperplasia (HHP), which developed in adolescence. We have reviewed reported familial cases of HHP and conclude that presentation is similar to sporadi cases and that all affected family members have been related through a maternal relative. PMID- 12900902 TI - Characterization of a Wilms tumor in a 9-year-old girl with trisomy 18. AB - This is a report of a trisomy 18 patient who developed Wilms tumor in conjunction with perilobar nephroblastomatosis (NB) at 9 years and 5 months of age. Review of the literature revealed that most patients with trisomy 18 who develop Wilms tumor, do so at a later than expected age for a tumor related to NB, and are females. In this case, no chromosome 11 WT1 mutation was detected by PCR/SSCP analysis, but the tumor had in addition to the trisomy, an isochromosome 7q and loss of heterozygosity at 16q, two mutations that have been linked independently to Wilms tumorigenesis. PMID- 12900903 TI - Preimplantation genetic diagnosis for a known cryptic translocation: follow-up clinical report and implication of segregation products. AB - This report describes preimplantation genetic diagnosis (PGD) of a couple with a known paternally-derived balanced cryptic translocation 46,XY.ish t(2q;17q)(210E14-,B37c1+;B37c1-,210E14+) in embryos from a couple who previously had a child with severe mental retardation and was previously described in this journal [Bacino et al., 2000]. This child inherited the unbalanced product of translocation from her father: 46,XX.ish der(2)t(2q;17q)pat(210E14-,B37c1+). The couple desired a normal offspring and sought PGD to avoid clinical pregnancy termination. They were treated three times with in vitro fertilization followed by PGD. Two sequential FISH hybridizations were performed. In the first hybridization, telomeric probes to 2q and 17q and a chromosome 17 centromere probe were employed. The second hybridization screened for maternal age-related aneuploidy (X,Y,13,18,21). Of the 18 informative embryos, only 4 (22%) were normal. The remaining 12 (67%) were abnormal; most with unbalanced products (10/12) from the paternally-derived rearrangement. The most frequent mode of segregation observed for this cryptic translocation was adjacent-1 (7/18, 39%). This suggests cryptic translocations are amenable to PGD and, as are traditional translocations, demonstrate higher frequencies of unbalanced segregants than the empiric risk of 10-15% observed at amniocentesis or chorionic villus sampling. Thus, cryptic translocations presumably behave like overt translocations, in that PGD must be performed on a relatively large number of embryos to assure even 2-3 transferable embryos. PMID- 12900904 TI - De novo paracentric inversion (X)(q26q28) with features mimicking Prader-Willi syndrome. AB - Different genetic and non-genetic disorders, including several chromosomal abnormalities, may mimic Prader-Willi syndrome (PWS). We report on an 11-year-old girl with features reminiscent of PWS due to an unreported de novo paracentric inversion Xq26q28. Microdeletion 15q11-q13 and maternal uniparental disomy 15 were ruled out. The importance of chromosomal studies in addition to molecular analysis on patients with features suggestive of PWS is stressed. PMID- 12900905 TI - Gorlin syndrome with ulcerative colitis in a Japanese girl. AB - We present the case of a 14-year-old Japanese girl who had both Gorlin syndrome and ulcerative colitis. She had complained of blood stools for 6 months and severe scoliosis from her infancy. Physical examination revealed multiple nevi, palmar and plantar pits, jaw cysts, and calcification of the falx cerebri, leading to the diagnosis of Gorlin syndrome. Total colonoscopy revealed an edematous and spotty bleeding mucosa extending from the anus to the transverse colon. Histological examination was also compatible with ulcerative colitis. Thus, we diagnosed her as having Gorlin syndrome with ulcerative colitis. Gene analysis revealed a mutation, 1247InsT, in the human patched gene (PTCH), resulting in the truncation of PTCH protein. Since Gorlin syndrome and ulcerative colitis are rare disorders in childhood, this association is interesting, suggesting a correlation between the hedgehog signaling and intestinal disorders. PMID- 12900906 TI - An acceptor splice site mutation in HOXD13 results in variable hand, but consistent foot malformations. AB - HOXD13 is the most 5' of the HOXD cluster of homeobox genes in chromosome band 2q31.1. Heterozygous expansions of a polyalanine tract in HOXD13 are typically associated with synpolydactyly characterized by insertional digit duplication associated with syndactyly. We screened for mutations of HOXD13 in patients with a variety of limb malformations and identified a novel heterozygous mutation (758 2delA) in a three-generation family without the typical synpolydactyly phenotype in the hands, but with bilateral partial duplication of the 2nd metatarsals within the first web space of the feet. This mutation locates in the acceptor splice site of exon 2 and is predicted to cause failure of normal splicing of HOXD13. The foot abnormality in this family is similar to that described in two families by Goodman et al. [1998: Am. J. Hum. Genet. 63: 992-1000] in which different deletions of HOXD13 were reported. These findings together lend support to a distinct phenotype resulting from haploinsufficiency of HOXD13. PMID- 12900907 TI - Dorfman-Chanarin syndrome in Egypt. PMID- 12900908 TI - Truncus arteriosus and duplication 8q. PMID- 12900909 TI - No mutation in the gene for Noonan syndrome, PTPN11, in 18 patients with Costello syndrome. PMID- 12900910 TI - Chromosome bands and ends revisited. PMID- 12900911 TI - Supernumerary digital flexion creases. PMID- 12900913 TI - Somatosensation in the superior colliculus of the star-nosed mole. AB - The superior colliculus (or optic tectum in nonmammals) plays a critical role in the visual system and is essential for integrating sensory inputs to guide eye and head movements. However, what is the role of the superior colliculus (SC) in species that depend almost exclusively on touch? In this study we examined the SC of the star-nosed mole, a subterranean mammal that, instead of using vision, explores its environment using its tactile star. The star acts like a mechanosensory eye with a central tactile fovea that is constantly shifted in a saccadic manner. Multiunit microelectrode recordings were used to determine the topography and receptive field organization of somatosensory inputs to the SC and to test for visual and auditory responses. Here we report an SC dominated by somatosensory inputs in which neurons in all layers responded to mechanosensory stimulation, forming a topographic representation of contralateral body dominated by the mechanosensory star. Receptive fields were large, and appendage representations overlapped, suggesting that the SC may use a distributed, population code to guide the saccadic movements of the mole's touch fovea. No auditory or visual responses were recorded from the SC, although neurons in the neighboring inferior colliculus responded to auditory stimuli. Layers IVb-VII were identified, and a layer superficial to IVb contained neurons that responded to somatosensory stimulation, suggesting that there are unique patterns of afferents in the star-nosed mole's SC. PMID- 12900914 TI - Rearrangement of synaptic connections with inhibitory neurons in developing mouse visual cortex. AB - Cortical inhibition is determined in part by the organization of synaptic inputs to gamma-aminobutyric acidergic (GABAergic) neurons. In adult rat visual cortex, feedforward (FF) and feedback (FB) connections that link lower with higher areas provide approximately 10% of inputs to parvalbumin (PV)-expressing GABAergic neurons and approximately 90% to non-GABAergic cells (Gonchar and Burkhalter [1999] J. Comp. Neurol. 406:346-360). Although the proportions of these targets are similar in both pathways, FF synapses prefer larger PV dendrites than FB synapses, which may result in stronger inhibition in the FF than in the FB pathway (Gonchar and Burkhalter [1999] J. Comp. Neurol. 406:346-360). To determine when during postnatal (P) development FF and FB inputs to PV and non-PV neurons acquire mature proportions, and whether the pathway-specific distributions of FF and FB inputs to PV dendrites develop from a similar pattern, we studied FF and FB connections between area 17 and the higher order lateromedial area (LM) in visual cortex of P15-42 mice. We found that the innervation ratio of PV and non-PV neurons is mature at P15. Furthermore, the size distributions of PV dendrites contacted by FF and FB synapses were similar at P15 but changed during the third to sixth postnatal weeks so that, by P36-42, FF inputs preferred thick dendrites and FB synapses favored thin PV dendrites. These results suggest that distinct FF and FB circuits develop after eye opening by rearranging the distribution of excitatory synaptic inputs on the dendritic tree of PV neurons. The purpose of this transformation may be to adjust differentially the strengths of inhibition in FF and FB circuits. PMID- 12900916 TI - Regenerating crayfish motor axons assimilate glial cells and sprout in cultured explants. AB - Phasic and tonic motor nerves originating from crayfish abdominal ganglia, in 2-3 day-old cultured explants, display at their transected distal ends growth zones from which axonal sprouts arise. The subcellular morphology of this regenerative response was examined with thin serial-section electron microscopy and reveals two major remodeling features. First, the external sprouts that exit the nerve are a very small part of a much more massive sprouting response by individual axons comprising several orders of internal sprouts confined to the nerve. Both internal and external sprouts have a simple construction: a cytoskeleton of microtubules and populations of mitochondria, clear synaptic vesicles, membranous sacs, and extrasynaptic active zone dense bars, features reminiscent of motor nerve terminals. Close intermingling of the sprouts of several axons give rise to a neuropil-like arbor within the nerve. Thus, extensive sprouting is an intrinsic response of crayfish motor axons to transection. Second, an equally dramatic remodeling feature is the appearance of nuclei, which resemble those of adjacent glial cells, within the motor axons. These nuclei often appear where the adjoining membranes of the axon and glial cell are disrupted and where free standing lengths of the double membrane are present. These images signify a breakdown of the dividing membranes and assimilation of the glial cell by the axon, the nucleus being the most visible sign of such assimilation. Thus, crayfish motor axons respond to transection by assimilating glial cells that may provide regulatory and trophic support for the sprouting response. PMID- 12900915 TI - ERMs colocalize transiently with L1 during neocortical axon outgrowth. AB - L1 is a member of the Ig superfamily of cell adhesion molecules (CAMs) that functions in many aspects of neuronal development including axonal outgrowth and neuronal migration. These functions require coordination between L1 and the actin cytoskeleton. Because CAMs and the cytoskeleton do not bind directly, membrane cytoskeletal linkers (MCLs) such as ankyrin are thought to be crucial to their interactions, but data from a knockout mouse suggest that ankyrin is not necessary for the earliest events attributed to L1 function. Recent findings in hippocampal cell culture show that members of the ERM family of proteins (ezrin, radixin, and moesin) can also serve as MCLs between L1 and actin in neurons. Here, we demonstrate that ERM proteins are expressed in extending neuronal processes in the intermediate zone of the developing cortex, a region that is densely packed with migrating neurons and growing axons. ERMs and L1 are codistributed extensively over a transient time course that coincides with rapid axon growth and cortical expansion. This codistribution is strong at embryonic day 17 and 19 but diminishes by postnatal day 0, at which time ankyrin-L1 codistribution increases dramatically. These findings suggest that in the developing neocortex, ERMs are the predominant MCL for L1 during migration and axon extension, neither of which requires ankyrin function. Furthermore, these data suggest that there is a developmentally regulated switch in MCL function in the developing brain. PMID- 12900917 TI - Temporal and spatial distribution of activated caspase-3 after subdural kainic acid infusions in rat spinal cord. AB - The molecular events initiating apoptosis following traumatic spinal cord injury (SCI) remain poorly understood. Soon after injury, the spinal cord is exposed to numerous secondary insults, including elevated levels of glutamate, that contribute to cell dysfunction and death. In the present study, we attempted to mimic the actions of glutamate by subdural infusion of the selective glutamate receptor agonist, kainic acid, into the uninjured rat spinal cord. Immunohistochemical colocalization studies revealed that activated caspase-3 was present in ventral horn motor neurons at 24 hours, but not 4 hours or 96 hours, following kainic acid treatment. However, at no time point examined was there evidence of significant neuronal loss. Kainic acid resulted in caspase-3 activation in several glial cell populations at all time points examined, with the most pronounced effect occurring at 24 hours following infusion. In particular, caspase-3 activation was observed in a significant number of oligodendroglia in the dorsal and ventral funiculi, and there was a pronounced loss of oligodendroglia at 96 hours following treatment. The results of these experiments indicate a role for glutamate as a mediator of oligodendroglial apoptosis in traumatic SCI. In addition, understanding the apoptotic signaling events activated by glutamate will be important for developing therapies targeting this cell death process. PMID- 12900918 TI - Ultrastructure and synaptic targets of tectothalamic terminals in the cat lateral posterior nucleus. AB - The recent appreciation of the fact that the pulvinar and lateral posterior (LP) nuclei receive two distinct types of cortical input has sparked renewed interest in this region of the thalamus. A key question is whether the primary or "driving" inputs to the pulvinar/LP complex originate in cortical or subcortical areas. To begin to address this issue, we examined the synaptic targets of tectothalamic terminals within the LP nucleus. Tectothalamic terminals were labeled using the anterograde transport of biotinylated dextran amine (BDA) or Phaselous leucoagglutinin placed in the superior colliculus or using immunocytochemical staining for substance P, a neurotransmitter found to be used by the tectothalamic pathway (Hutsler and Chalupa [ 1991] J. Comp. Neurol. 312:379-390). Our results suggest that most tectothalamic terminals are large and occupy a proximal position on the dendritic arbor of LP relay cells. In the medial LP, tectothalamic terminals labeled by the transport of neuronal tracers or substance P immunocytochemistry can form tubular clusters that surround the proximal dendrites of relay cells. In a rostral and lateral subdivision of the lateral LP nucleus (LPl-2), tectothalamic terminals form more typical glomerular arrangements. When compared with existing physiological data, these results suggest that a unique integration of tectal and cortical inputs may contribute to the response properties of LP neurons. PMID- 12900919 TI - Xenopus laevis CB1 cannabinoid receptor: molecular cloning and mRNA distribution in the central nervous system. AB - In the present research we isolated and characterized Xenopus laevis CB1 cannabinoid receptor mRNA. The CB1 coding sequence shows a high degree of identity with those of other vertebrates, mammals included, confirming that CB1 receptor is conserved over the course of vertebrate evolution. Notably, the similarity between the X. laevis CB1 sequence and that of the urodele amphibian Taricha granulosa is not higher than the similarity existing between Xenopus and mammals, thus supporting phylogenetic distance between anurans and urodeles. By means of in situ hybridization histochemistry, CB1 mRNA expression and distribution was investigated in the X. laevis central nervous system. As revealed, CB1 mRNA-containing neurons are numerous in the prosencephalon, especially in the olfactory bulbs, telencephalic pallium, and hypothalamus. In the midbrain and hindbrain, labeled cells were observed in the mesencephalic tegmentum and dorsolateral romboencephalon. Abundant CB1 mRNA positive neurons are localized throughout the gray matter of the spinal cord, in particular in the dorsal and ventral fields, where labeled motor neurons are also observed. The distribution of CB1 mRNA in the Xenopus CNS is generally consistent with the CB1 like-immunohistochemistry results we have previously obtained, showing in amphibians a well developed cannabinergic system almost comparable to that described in mammals. However, some differences, such as the abundance of CB1 mRNA-containing neurons in the olfactory system and the rich CB1 spinal innervation, are found. PMID- 12900920 TI - GABA- and glycine-like immunoreactivity in axons and dendrites contacting the central terminals of rapidly adapting glabrous skin afferents in rat spinal cord. AB - The object of the present study was to determine the nature and distribution of synaptic contacts on the terminals of rapidly adapting mechanosensory afferents innervating the glabrous skin of the rat foot. Afferents were physiologically characterized by intracellular recording, before injection with neurobiotin and preparation for electron microscopy. Axon terminals were serially sectioned and immunolabeled with antibodies against GABA and glycine using a postembedding immunogold method. Afferent boutons in lamina III were often surrounded by several presynaptic axons and postsynaptic dendrites (thus forming type II glomeruli), while boutons in laminae IV-V had only simple, nonglomerular interactions. In both regions triadic synaptic arrangements where presynaptic interneurons contact both afferent boutons and their postsynaptic dendrites were present in 50-75% of boutons. Approximately three-quarters of presynaptic axons were immunoreactive for both GABA and glycine and most of the remainder for GABA alone. Most postsynaptic dendrites were not immunoreactive. Comparisons are made with information from similar studies of other rat and cat afferents conducting in the Aalphabeta range. This demonstrates that although the principles of control may be similar for cutaneous afferents of this type there are significant differences between cutaneous and 1a muscle afferents in the rat. There are also differences in detail between the interactions of afferents of the same modality in rat and cat; in the rat there are greater numbers of presynaptic axons per bouton and a greater proportion of boutons receive axo-axonic contacts and are involved in synaptic triads. PMID- 12900921 TI - Development of output connections from the inferior colliculus to the optic tectum in barn owls. AB - We studied the development of the projection from the external nucleus of the inferior colliculus (ICX) to the optic tectum (OT) in the barn owl. The projection was labeled by tracer application in vitro to either the OT or the ICX, or by staining ICX cells intracellularly with biocytin. The axons of ICX neurons bifurcated into an ascending branch that projected toward the OT and a descending branch that coursed caudally to an unknown target in the brainstem. Axons of the ICX were observed to grow into the OT from embryonic day 16 (E16) on. From E22 on, side branches of the axonal projections could be found within the OT. At the day of hatching (E32), the projection displayed a dorsoventral topography comparable to the adult owl; however, atopically projecting cells remained. The complexity of the axonal arborization in the adult barn owl was found to be slightly increased compared with the hatchling. The terminal area of individual ICX cells in the OT of the adult barn owl was still broad, a finding that had not been expected from the sharply defined physiological response properties of the bimodal neurons in the space map of the OT. However, the width of the termination zone was in accordance with the large dendritic tree of the adult ICX cells, because both spanned comparable angles in their respective maps. Our data suggest that a coarse projection from the ICX to the OT can develop without coherent sensory input and may, therefore, be innately determined. PMID- 12900922 TI - Synaptic and morphologic properties in vitro of premotor rat nucleus tractus solitarius neurons labeled transneuronally from the stomach. AB - Neurons in the rat nucleus tractus solitarius (NTS) possess morphologic characteristics that have been correlated with the type of synaptic information they receive. These features have been described for viscerosensory neurons but not for premotor NTS neurons. The morphologic and synaptic features of neurons in the rat caudal NTS were assessed using whole-cell patch-clamp recordings and biocytin labeling in brainstem slices. Gastric-related premotor NTS neurons were identified for recording after inoculation of the stomach wall with a transneuronal retrograde viral label that reports enhanced green fluorescent protein. Three morphologic groups of NTS neurons were identified based on quantitative aspects of soma area and proximal dendritic arborization, measures that were consistent across slice recordings. The most common type of cell (group I) had relatively small somata and one to three sparsely branching dendrites, whereas the other groups had larger somata and more than three dendrites, which branched predominantly close to (group II) or distant from (group III) the soma. Voltage-clamp recordings revealed spontaneous excitatory and inhibitory postsynaptic currents in all neurons, regardless of morphology. Gastric-related premotor NTS neurons composed two of the three morphologic types (i.e., groups I and II). Compared with unlabeled neurons, these cells were less likely to receive constant-latency synaptic input from the tractus solitarius. These results refute the hypothesis that general patterns of synaptic input to NTS neurons depend on morphology. Gastric premotor neurons comprise a subset of NTS morphologic types, the organization of the viscerosensory input to which has yet to be defined. PMID- 12900923 TI - Organization of glomeruli in the main olfactory bulb of Xenopus laevis tadpoles. AB - Structural and functional investigations were carried out to study olfactory glomeruli in the main olfactory bulb (OB) in tadpoles of the clawed frog, Xenopus laevis. Calcium imaging of odor response patterns of OB neurons revealed that the synapses within the glomeruli are functional. Tracing axons of individual olfactory receptor neurons (ORNs), dendrites of mitral/tufted (M/T) cells and processes of periglomerular interneurons indicate that the glomerular architecture is solely determined by terminal branches of ORN axons and tufts of M/T primary dendrites. The small population of periglomerular neurons forms wide field arborizations that always extend over many glomeruli, enter the glomeruli, but lack any glomerular tufts. Antibodies to synaptophysin indicate a high density of synapses within glomeruli, which was further confirmed at the ultrastructural level and quantified to approximately 0.5 synaptic sites per microm(2). Combining immunocytochemistry and ultrastructural investigations, we show that glomeruli in Xenopus laevis tadpoles lack any cellular borders. Glomeruli are surrounded neither by periglomerular somata nor by glial processes. Taken together, our results demonstrate that olfactory glomeruli in Xenopus laevis tadpoles (1) are fully functional, (2) are spheroidal neuropil aggregations of terminal tufts of ORNs and tufts of primary dendrites of M/T cells, and (3) are not enwrapped by a border formed by juxtaglomerular cells. PMID- 12900924 TI - Patterns of PERIOD and pigment-dispersing hormone immunoreactivity in the brain of the European honeybee (Apis mellifera): age- and time-related plasticity. AB - We explored the neural basis of age- and task-related plasticity in circadian patterns of activity in the honeybee. To identify putative circadian pacemakers in the bee brain, we used antibodies against Drosophila melanogaster and Antheraea pernyi PERIOD and an antiserum to crustacean pigment-dispersing hormone (PDH) known to cross-react with insect pigment-dispersing factors (PDFs). In contrast to previous results from Drosophila, PDH and PER immunoreactivity (-ir) were not colocalized in bee neurons. The most intense PER-ir was cytoplasmic, in two groups of large neurons in the protocerebrum. The number of protocerebral PER ir neurons and PER-ir intensity within individual cells were highest in brains collected during subjective night and higher in old bees than in young bees. These results are consistent with previous analyses of brain per mRNA in honeybees. Nuclear PER-ir was found throughout the brain, including the optic and antennal lobes. A single group of PDH-ir neurons (approximately 20/optic lobe) was consistently and intensely labeled at the medial margin of the medulla, independent of age or time of day. The processes of these neurons extended to specific neuropils in the protocerebrum and the optic lobes but not to the deutocerebrum. The patterns displayed by PER- and PDH-ir do not completely match any patterns previously described. This suggests that, although clock proteins are conserved across insect groups, there is no universal pattern of coexpression that allows ready identification of pacemaker neurons within the insect brain. PMID- 12900925 TI - Comparative distribution of neuropeptide Y Y1 and Y5 receptors in the rat brain by using immunohistochemistry. AB - Neuropeptide Y (NPY) Y1 and Y5 receptor subtypes mediate many of NPY's diverse actions in the central nervous system. The present studies use polyclonal antibodies directed against the Y1 and Y5 receptors to map and compare the relative distribution of these NPY receptor subtypes within the rat brain. Antibody specificity was assessed by using Western analysis, preadsorption of the antibody with peptide, and preimmune serum controls. Immunostaining for the Y1 and Y5 receptor subtypes was present throughout the rostral-caudal aspect of the brain with many regions expressing both subtypes: cerebral cortex, hippocampus, hypothalamus, thalamus, amygdala, and brainstem. Further studies using double label immunocytochemistry indicate that Y1R immunoreactivity (-ir) and Y5R-ir are colocalized in the cerebral cortex and caudate putamen. Y1 receptor ir was evident in the central amygdala, whereas both Y1- and Y5-immunoreactive cells and fibers were present in the basolateral amygdala. Corresponding with the physiology of NPY in the hypothalamus, both Y1R- and Y5R-ir was present within the paraventricular (PVN), supraoptic, arcuate nuclei, and lateral hypothalamus. In the PVN, Y5R-ir and Y1R-ir were detected in cells and fibers of the parvo- and magnocellular divisions. Intense immunostaining for these receptors was observed within the locus coeruleus, A1-5 and C1-3 nuclei, subnuclei of the trigeminal nerve and nucleus tractus solitarius. These data provide a detailed and comparative mapping of Y1 and Y5 receptor subtypes within cell bodies and nerve fibers in the brain which, together with physiological and electrophysiological studies, provide a better understanding of NPY neural circuitries. PMID- 12900926 TI - Somatotopic mapping of chordotonal organ neurons in a primitive ensiferan, the New Zealand tree weta Hemideina femorata: I. femoral chordotonal organ. AB - The femoral chordotonal organ (FCO) in orthopteran insects comprises several hundred sensory neurons, making it one of the most complex insect proprioceptors. The sensory neurons are suspended from the proximal femur, connecting distally to ligaments and to a needle-like apodeme extending from the proximal tibia. They monitor the position and movement of the tibia. To address how this complexity depends on evolutionary status and function, the morphology of the FCO neurons in the primitive orthopteran Hemideina femorata was investigated by staining small populations of identified afferents. As in crickets, the FCOs in all legs of the weta comprise partly fused ventral and dorsal scoloparia, with the former containing two groups of somata, the ventral group (VG) and the dorsal group (DG). However, the dendrites of the DG insert into thin connective tissue attached to the ventral side of the dorsal ligament, forming a "third scoloparium." The VG afferents terminate mainly in the motor association neuropils, whereas afferents from the dorsal scoloparium neurons terminate exclusively in the vibratory neuropil as do the afferents from the subgenual organ, a substrate vibration detector. Several afferents originating in the DG have extensive terminations in the motor association-, vibratory-, and auditory processing neuropils, indicating lesser functional specialization than in the other groups. The evolutionary development of the FCO is discussed from a comparative viewpoint. PMID- 12900927 TI - Somatotopic mapping of chordotonal organ neurons in a primitive ensiferan, the New Zealand tree weta Hemideina femorata: II. complex tibial organ. AB - Most ensiferan insects possess sets of highly specialized chordotonal organs in the proximal tibiae to detect conspecific auditory/vibratory signals or approach of predators. To date, most auditory/vibratory afferents have been classified according to their physiological properties and axonal projection morphology, but not to somatotopic origins. Hence, the functional specialization of identified receptor cells in the tibial organs remains uncertain. To address this question from an anatomical aspect, we investigated the structure of the weta, Hemideina femorata, tibial organs (the most elaborated tibial chordotonal organs among ensiferans) and their central projections by staining small numbers of receptor afferents from identified tibial organs. These organs comprise the "complex tibial organ," including the subgenual organ (primary vibration detector) and its posterior complement, the accessory organ, and the crista acustica (primary auditory organ) and its proximal complement, the intermediate organ. Unlike reports of a membranous organ structure for homologs in other ensiferans, weta tibial organs contain receptor cells embedded in thick solid masses. Primary afferents project ipsilaterally to the medial ventral association center of thoracic ganglia, where axon terminals are arrayed topographically in different areas specific to each organ, except for almost complete overlap of afferents originating from the distal part of the crista acustica and from the intermediate organ. In contrast to somatotopic reflection of sensilla position on limbs, as known for mechanoreceptor hairs, the somatotopic projection map of the insect ear reveals topographic association with acoustic tracheae or tibial cuticular attachment sites, which in turn must reflect determinants of response sensitivity (e.g., frequency or threshold). PMID- 12900928 TI - Spontaneous regeneration of the corticospinal tract after transection in young rats: collagen type IV deposition and astrocytic scar in the lesion site are not the cause but the effect of failure of regeneration. AB - In young rats the corticospinal tract regenerated after a single transection of the spinal cord with a sharp blade, but regeneration failed if the transection was repeated to make a more traumatic injury. To identify cells and associated molecules that promote or impede regeneration, we compared expression of collagen type IV, glial fibrillary acidic protein (GFAP), and vimentin immunoreactivity (IR) at the lesion sites in combination with anterograde axonal tracing between animals with two types of transection. Axonal regeneration occurred as early as 18 hours after transection; regenerating axons penetrated vessel-like structures with collagen type IV-IR at the lesion site, while reactive astrocytes coexpressing GFAP- and vimentin-IR appeared in the lesioned white matter. In contrast, when regeneration failed astrocytes were absent near the lesion. By 7 days sheet-like structures with collagen type IV-IR and astrocytic scar appeared in the lesioned white matter and persisted until the end of the observation period (31 days). On the basis of their spatiotemporal appearance, collagen type IV-IR sheet-like structures and the astrocytic scar follow, rather than cause, the failure of regeneration. The major sign, and perhaps cause, of failure of axonal regeneration is likely the prolonged disappearance of astrocytes around the lesion site in the early postinjury period. PMID- 12900930 TI - Spatial patterns of mammalian brain aging: distribution of cathepsin D immunoreactive cell bodies and dystrophic dendrites in aging dogs resembles that in Alzheimer's disease. AB - Elevated levels of the lysosomal enzyme cathepsin D are found in the early stages of Alzheimer's disease (AD) and co-occur with intraneuronal tangles. The present study tested whether increases in cathepsin D would emerge during aging in another mammalian species. Regional brain patterns of cathepsin D immunostaining were compared in dogs ages 0.35 to 16 years. Accumulations of immunopositive material were evident in neuronal cell bodies in many forebrain sites in middle age to old dogs (>/=6 years). Three types could be distinguished: (1) dense aggregates with no particular position within the cell body; (2) crescent-shaped "caps" that occupied one pole of the cell body; and (3) very dense "spikes" that extended from the cell body for variable distances into the apical dendrite; these spikes were found in only a few areas, most notably the subiculum and layer V of neocortex. The spikes appeared between ages 2 and 5 years and increased steadily with age thereafter. Spikes were found in the subiculum in the aged human brain but only infrequently; they were, however, present in large numbers in AD brains. These results established that brain aging in dogs is (1) well advanced by middle age, (2) varies markedly across regions, and (3) in at least some of its aspects (dystrophic dendrites) is prominent in areas known to exhibit pathology early in the course of AD. Combined with previous results for rats, these findings indicated that changes in cathepsin D observed in AD, in particular in the temporal lobe, reflect a generalized mammalian pattern of brain aging. PMID- 12900931 TI - Increased expression of insulin-like growth factor I augments the progressive phase of synaptogenesis without preventing synapse elimination in the hypoglossal nucleus. AB - The in vivo actions of insulin-like growth factor I (IGF-I) on synaptogenesis in the hypoglossal nucleus were investigated in transgenic mice that overexpress IGF I in the brain postnatally and in normal nontransgenic littermate controls. In a previous study using these mice, we found that IGF-I increases the total volume of the hypoglossal nucleus by increasing the volume of neuropil rather than by increasing total neuron number; therefore, the progressive and regressive phases of synaptogenesis could be evaluated without the confounding effects of altered neuron number. The volume of the hypoglossal nucleus was significantly increased by 28% to 59% in transgenic mice after postnatal day (P) 7, whereas the total number of hypoglossal neurons did not differ significantly from controls. The numerical density of neurons was significantly decreased by 21% to 38% after P7, and the density of myelinated axons was significantly increased by 19%. Although the numerical density of synapses did not differ between groups at any age, the total number of synapses in transgenic mice was increased by 42% to 52% after P14. Total synapse number in controls increased from P7 (7.9 million) to peak values at P21 (36.0 million), followed by a significant decrease (33%) at P130 (24.2 million). In transgenic mice, total synapses increased from 8.2 million on P7 to 51.1 million on P21, followed by a significant decrease (28%) to 36.7 million at P130. Our results demonstrated that IGF-I can stimulate a persistent increase in the number of hypoglossal synapses, thereby augmenting the progressive phase of synaptogenesis without preventing synapse elimination during the regressive phase. PMID- 12900929 TI - Connexin29 and connexin32 at oligodendrocyte and astrocyte gap junctions and in myelin of the mouse central nervous system. AB - The cellular localization, relation to other glial connexins (Cx30, Cx32, and Cx43), and developmental expression of Cx29 were investigated in the mouse central nervous system (CNS) with an anti-Cx29 antibody. Cx29 was enriched in subcellular fractions of myelin, and immunofluorescence for Cx29 was localized to oligodendrocytes and myelinated fibers throughout the brain and spinal cord. Oligodendrocyte somata displayed minute Cx29-immunopositive puncta around their periphery and intracellularly. In developing brain, Cx29 levels increased during the first few postnatal weeks and were highest in the adult brain. Immunofluorescence labeling for Cx29 in oligodendrocyte somata was intense at young ages and was dramatically shifted in localization primarily to myelinated fibers in mature CNS. Labeling for Cx32 also was localized to oligodendrocyte somata and myelin and absent in Cx32 knockout mice. Cx29 and Cx32 were minimally colocalized on oligodendrocytes somata and partly colocalized along myelinated fibers. At gap junctions on oligodendrocyte somata, Cx43/Cx32 and Cx30/Cx32 were strongly associated, but there was minimal association of Cx29 and Cx43. Cx32 was very sparsely associated with astrocytic connexins along myelinated fibers. With Cx26, Cx30, and Cx43 expressed in astrocytes and Cx29, Cx32, and Cx47 expressed in oligodendrocytes, the number of connexins localized to gap junctions of glial cells is increased to six. The results suggested that Cx29 in mature CNS contributes minimally to gap junctional intercellular communication in oligodendrocyte cell bodies but rather is targeted to myelin, where it, with Cx32, may contribute to connexin-mediated communication between adjacent layers of uncompacted myelin. PMID- 12900932 TI - Developmental expression pattern of monoamine oxidases in sensory organs and neural crest derivatives. AB - Serotonin (5-HT) has been shown to act as a morphogen in craniofacial and heart development and in the migration of neural crest derivatives. Some of these structures are capable of capturing 5-HT during development, but nothing is known about the localization of the main monoamine degradation enzymes, monoamine oxidase (MAO) A and B, in these developing tissues. We generated a highly specific antibody to MAOB; immunoreactivity is entirely abolished in brain extracts or brain sections of mice lacking MAOB. From the use of this antibody and specific riboprobes, we report that MAOB is expressed early in a variety of neural crest derivatives, in facial sensory organs, and in the heart. From E11.5 to P0, MAOB was found to be strongly expressed in the following neural crest derivatives: the aorta, cranial mesenchyme (developing bones, sensory neurons of the cranial ganglia, cartilages, thyroid, and striate muscles), dental mesenchyme, several soft palate derivatives, and boundary cap cells (E11.5-P4). Boundary cap cells contribute to the formation of nerve exit-entry points between the central and the peripheral nervous systems. Several facial sensory organs also contained MAOB mRNA, protein, and activity. High MAOB expression was noted in the olfactory placode, the dorsal part of the olfactory epithelium, the olfactory nerve layer (probably the ensheathing glia), the cochlear ganglionic cells, the taste buds, and the Merkel cells in the vibrissae follicles. Finally, we found that MAOB is massively expressed in the pharyngeal organ, heart, liver, and mast cells. In contrast, MAOA expression was restricted to the sympathetic ganglia and to the meningeal and capillary blood vessels. The pattern of MAOB expression generally matched the previously reported patterns of expression of the plasma 5-HT transporter expression or of the histamine biosynthetic enzyme L histidine decarboxylase, suggesting a role for MAOB in fine regulation of the levels of 5-HT and histamine in the developing embryo. PMID- 12900933 TI - Distribution of transient receptor potential channels in the rat carotid chemosensory pathway. AB - Glomus cells in the carotid body respond to decreases in oxygen tension of the blood and transmit this sensory information in the carotid sinus nerve to the brain via neurons in the petrosal ganglion. G-protein-coupled membrane receptors linked to phospholipase C may play an important role in this response through the activation of the cation channels formed by the transient receptor potential (TRP) proteins. In the present study, expression of TRPC proteins in the rat carotid body and petrosal ganglion was examined using immunohistochemical techniques. TRPC3, TRPC4, TRPC5, TRPC6, and TRPC7 were present in neurons throughout the ganglion. TRPC1 was expressed in only 28% of petrosal neurons, and of this population, 45% were tyrosine hydroxylase (TH)-positive, accounting for essentially all the TH-expressing neurons in the ganglion. Because TH-positive neurons project to the carotid body, this result suggests that TRPC1 is selectively associated with the chemosensory pathway. Confocal images through the carotid body showed that TRPC1/3/4/5/6 proteins localize to the carotid sinus nerve fibers, some of which were immunoreactive to an anti-neurofilament (NF) antibody cocktail. TRPC1 and TRPC3 were present in both NF-positive and NF negative fibers, whereas TPRC4, TRPC5, and TRPC6 expression was primarily localized to NF-negative fibers. Only TRPC1 and TRPC4 were localized in the afferent nerve terminals that encircle individual glomus cells. TRPC7 was not expressed in sensory fibers. All the TRPC proteins studied were present in type I glomus cells. Although their role as receptor-activated cation channels in the chemosensory pathway is yet to be established, the presence of TRPC channels in glomus cells and sensory nerves of the carotid body suggests a role in facilitating and/or sustaining the hypoxic response. PMID- 12900934 TI - Centrifugal partition chromatography: a survey of its history, and our recent advances in the field. AB - Centrifugal partition chromatography (CPC) was introduced in 1982 by Sanki Eng. And for almost 20 years this company remained the only one manufacturing such an apparatus. A CPC instrument or a CPC column is a series of channels linked in cascade by ducts and aligned in cartridges or disks in a circle around a rotor; setting the rotor in motion submits this assembly to a constant centrifugal field. The originality of CPC is that it uses any biphasic liquid-liquid system as mobile and stationary phases. The United States branch of Sanki contributed greatly to the worldwide acceptance of the technique. Recent works performed in the Netherlands and in France have, by means of visualization of flow-patterns in CPC channels, contributed to a better knowledge of hydrodynamics and mass transfer phenomena. Nowadays research in our laboratory focuses on speediness and scale-up of the technique. PMID- 12900935 TI - New reactions of 1-alkynes catalyzed by transition metal complexes. AB - Recently discovered catalytic reactions with ruthenium and lanthanide metal complexes have extended the scope of 1-alkynes as useful reagents. The specific formation of aryl-substituted (Z)-1,3-enzymes via the dimerization of HC(triple bond) CR(1) (R(1) = aryl) has been attained using dimeric lanthanide complexes, the catalytic activity of which appears to be unaffected by time. The dimerization of HC(triple bond) CR(2) (R(2) = t-Bu, SiMe(3)) catalyzed by Ru(cod)(cot)/PR(3) or RuH(2)(PPh(3))(3) produces a good yield of butatrienes (Z)R(2)CH=C=C=CHR(2) with a high degree of selectivity. Under certain conditions, HC(triple bond) C=SiMe(3) dimerizes to yield exclusively (Z)-M(3)Si-C(triple bond) C-CH=CH-SiMe(3). The hydration of HC(triple bond)CR(3) (R(3) = alkyl, aryl) catalyzed by RuCl(2)/PR'(3) or CpRuCl(PR"(3))(2) has realized the first example of anti-Markovnikov regioselectivity in an addition reaction of water that produces aldehydes R(3)CH(2)bond;CHO. The application of this reaction to propargylic alcohols has lead to their formal isomerization to alpha,beta unsaturated aldehydes. In contrast, the addition of amines R(4)bond;NH(2) (R(4) = aryl) to HCtbond;CR(5) (R(5) = alkyl, aryl) conforms to Markovnikov's rule to produce ketimines R(5)bond;(C=NR(4))bond;CH(3) when catalyzed by a Ru(3)(CO)(12)/additive. Since the reaction can be performed in air without the need for any solvents, it enables the practical synthesis of aromatic ketimines, which are difficult to prepare by conventional methods. The synthesis of indoles using deactivated anilines is one practical application of this reaction. The mechanisms of some of these reactions have been analyzed in detail with the aid of theoretical calculations. PMID- 12900936 TI - Microbial genes and enzymes in the degradation of chlorinated compounds. AB - Microorganisms are well known for degrading numerous natural compounds. The synthesis of a multitude of chlorinated compounds by the chemical industry and their release into the natural environment have created major pollution problems. Part of the cause of such pollution is the inability of natural microorganisms to efficiently degrade synthetic chlorinated compounds. Microorganisms are, however, highly adaptable to changes in the environment and have consequently evolved the genes that specify the degradation of chlorinated compounds to varying degrees. Highly selective laboratory techniques have also enabled the isolation of microbial strains capable of utilizing normally recalcitrant highly chlorinated compounds as their sole source of carbon and energy. The evolution and role of microbial genes and enzymes, as well as their mode of regulation and genetic interrelationships, have therefore been the subjects of intense study. This review emphasizes the genetic organization and the regulation of gene expression, as well as evolutionary considerations, regarding the microbial degradation of chlorobenzoates, chlorocatechols, and chlorophenoxyacetic acids. PMID- 12900938 TI - Cluster, facets, and edges: site-dependent selective chemistry on model catalysts. AB - More than activity, selectivity of catalytic reactions is the focus of research in the 21(st) century. We review studies on model systems that address the issue of directing a catalytic reaction on disperse metal catalysts by controlling the specific surface site. Three examples are explored: methanol dehydrogenation over Pd/alumina, NO dissociation on Pd/alumina, and reaction studies for molecules relevant in a Fischer-Tropsch scenario on a bimetallic Pd/Co/alumina model catalyst. We show how surface science can be used by combining a variety of experimental techniques to study the chemistry of model catalysts at the atomic level. PMID- 12900937 TI - Natural products of the medicinal fungus Ganoderma lucidum: occurrence, biological activities, and pharmacological functions. AB - Ganoderma lucidum, a fungus used in traditional Chinese medicine, produces polysaccharides and oxygenated triterpenoids with a very broad spectrum of biological activities and pharmacological functions. Among the Ganoderma triterpenoids, many pairs of C-3 alpha/beta stereoisomers and C-3/C-15 positional isomers have been identified. Biosynthetic study has indicated that the C-3alpha series of oxygenated triterpenoids is derived from the C-3beta series via an oxidation-reduction pathway. The interaction of Ganoderma triterpenoids with human platelets in the induction of aggregation and inhibition of agonist-induced aggregation and signal transduction has been elucidated. Reduction of cellular mevalonate content to a stage in which cholesterol synthesis is strongly inhibited and cell growth is marginally arrested sensitizes hepatoma cells to the oxygenated triterpenoids. A combination treatment of lovastatin and Ganoderma triterpenoids in animal studies has exhibited a potential anticancer effect. PMID- 12900939 TI - Effects of 17alpha-ethinylestradiol on immune parameters in the lizard Sceloporus occidentalis. AB - We examined the effect of 17alpha-ethinylestradiol on immunity of the Western fence lizard, Sceloporus occidentalis. Injection of 17alpha-ethinylestradiol resulted in dose-dependent suppression of peripheral blood leukocyte levels as determined by cell counts, whereas total spleen cell levels were decreased only at higher doses of 17alpha-ethinylestradiol. In contrast, spleen cell proliferation was enhanced by 17alpha-ethinylestradiol as measured by reduction of MTT to formazan following a two-way mixed lymphocyte reaction. Antibody responses were unaffected. Effects on peripheral blood leukocyte levels and spleen cell proliferation similar to those observed in response to injection of 17alpha-ethinylestradiol were observed following injection of a single dose of hydrocortisone. However, injection of lizards with 17alpha-ethinylestradiol did not result in a significant increase in serum cortisol. Results of this study suggest that exposure of Western fence lizards to 17alpha-ethinylestradiol leads to decreased numbers of circulating leukocytes and total spleen cell numbers and the enhancement of spleen cell proliferation in a two-way mixed lymphocyte reaction. These effects probably involve mechanisms other than or in addition to the induction of cortisol release. PMID- 12900940 TI - Thermal and metabolic factors affecting bioaccumulation of triazine herbicides by rainbow trout (Oncorhynchus mykiss). AB - The bioaccumulation of the triazine herbicides terbutryn and terbuthylazine in rainbow trout was investigated at water temperatures of 4 degrees C, 10 degrees C, and 17 degrees C. The synergetic effect of water temperature and herbicide on fish liver detoxification metabolism was also studied by measuring liver ethoxyresorufin-O-deethylase (EROD) and UDP-glucuronosyltransferase (UDPGT) activities. The bioaccumulation of terbutryn increased significantly with increasing water temperature. However, temperature seemed to have no effect on the bioaccumulation of terbuthylazine. Both chemicals decreased the measured enzyme activities. Also, temperature effects were significant. In all tests the highest liver EROD activities were measured in fish exposed to a water temperature of 4 degrees C, and the activities decreased with increasing water temperature. The thermal effects on fish liver UDPGT activity differed between the herbicides. The highest activities at a water temperature of 4 degrees C were measured in fish exposed to terbutryn, whereas fish exposed to terbuthylazine had the highest liver UDPGT activities at 17 degrees C. Measuring liver detoxification metabolism together with bioaccumulation increases the understanding of bioaccumulation of chemicals in different environmental conditions. PMID- 12900941 TI - Pulmonary responses of acute exposure to ultrafine iron particles in healthy adult rats. AB - As critical constituents of ambient particulate matter, transition metals such as iron may play an important role in health outcomes associated with air pollution. The purpose of this study was to determine the respiratory effects of inhaled ultrafine iron particles in rats. Sprague Dawley rats 10-12 weeks of age were exposed by inhalation to iron particles (57 and 90 microg/m(3), respectively) or filtered air (FA) for 6 h/day for 3 days. The median diameter of particles generated was 72 nm. Exposure to iron particles at a concentration of 90 microg/m(3) resulted in a significant decrease in total antioxidant power along with a significant induction in ferritin expression, GST activity, and IL-1beta levels in lungs compared with lungs of the FA control or of animals exposed to iron particles at 57 microg/m(3). NFkappaB-DNA binding activity was elevated 1.3 fold compared with that of control animals following exposure to 90 microg/m(3) of iron, but this change was not statistically significant. We concluded that inhalation of iron particles leads to oxidative stress associated with a proinflammatory response in a dose-dependent manner. The activation of NFkappaB may be involved in iron-induced respiratory responses, but further studies are merited. PMID- 12900942 TI - Effect of Microcystis aeruginosa and Nodularia spumigena on survival of Eurytemora affinis and the embryonic and larval development of the Baltic herring Clupea harengus membras. AB - Laboratory experiments were carried out to investigate the effect of two strains of Microcystis aeruginosa and a strain of Nodularia spumigena on the survival of Eurytemora affinis (Copepoda) and on the embryonic and larval development of the Baltic spring-spawning herring Clupea harengus membras. The trials were made in water taken from Parnu Bay, at a salinity of 3.7-5.1 psu, a constant temperature (15 degrees C +/- 1 degrees C in trials with Eurytemora and herring embryos; 18 degrees C +/- 2 degrees C with herring larvae), and an oxygen concentration of 8.8-10.4 ppm. The strains tested had a negative impact on the survival of Eurytemora, as well as on the embryonic development and hatching regime of the Baltic herring. In Eurytemora the response depended on the sex of the animals: the survival was clearly higher in females. In the embryonic stages of herring, the influence resulted in an increase in deviations from the normal pattern of development and a higher mortality. The impact of the strains on the larval development of herring was rather moderate. PMID- 12900943 TI - Cylindrospermopsin-induced protein synthesis inhibition and its dissociation from acute toxicity in mouse hepatocytes. AB - The toxicology of the cyanobacterial alkaloid cylindrospermopsin (CYN), a potent inhibitor of protein synthesis, appears complex and is not well understood. In exposed mice the liver is the main target for the toxic effects of CYN. In this study primary mouse hepatocyte cultures were used to investigate the mechanisms involved in CYN toxicity. The results show that 1-5 microM CYN caused significant concentration-dependent cytotoxicity (52%-82% cell death) at 18 h. Protein synthesis inhibition was a sensitive, early indicator of cellular responses to CYN. Following removal of the toxin, the inhibition of protein synthesis could not be reversed, showing behavior similar to that of the irreversible inhibitor emetine. In contrast to the LDH leakage, protein synthesis was maximally inhibited by 0.5 microM CYN. No protein synthesis occurred over 4-18 h at or above this concentration. Inhibition of cytochrome P450 (CYP450) activity with 50 microM proadifen or 50 microM ketoconazole diminished the toxicity of CYN but not the effects on protein synthesis. These findings imply a dissociation of the two events and implicate the involvement of CYP450-derived metabolites in the toxicity process, but not in the impairment of protein synthesis. Thus, the total abolition of protein synthesis may exaggerate the metabolite effects but cannot be considered a primary cause of cell death in hepatocytes over an acute time frame. In cell types deficient in CYP450 enzymes, protein synthesis inhibition may play a more crucial role in the development of cytotoxicity. PMID- 12900944 TI - Acute toxicity and bioaccumulation of arsenic in tilapia (Oreochromis mossambicus) from a blackfoot disease area in Taiwan. AB - The general objective of our work was to determine the acute toxicity and bioaccumulation of arsenic (As) in tilapia (Oreochromis mossambicus) from the blackfoot disease (BFD) area in Taiwan. The average concentration of As in pond water ranged from 17.8 to 49 microg L(-1). Acute toxicity tests showed that the As concentration that caused toxicity to tilapia ranged from 69 060 microg As L( 1), in the 24-h toxicity test, to 28 680 microg As L(-1), in the 96-h toxicity test. We measured As concentrations in various tissues of tilapia to identify the affinities of tissues for As. Significant correlations were found among the As concentrations in all tissues. The highest bioconcentration factor (BCF) was found in the intestine (maximum value: 2270). The order of BCFs was: intestine > stomach > liver approximately gill > muscle. Arsenic concentrations in all tissues were allometric, negatively correlating with fish body weight [r(2) = 0.63 +/- 0.045 (mean +/- SE), p < 0.05]. Our results also revealed that As concentrations in muscle tissue were positively correlated with As accumulation in the viscera (r(2) = 0.85, p < 0.05). Significantly higher concentrations of As were obtained in the viscera of tilapia [12.65 +/- 10.17 microg g(-1) dry wt (mean +/- SD)] than in the muscle tissue (3.55 +/- 0.42 microg g(-1) dry wt). Our results suggest that a simple way of reducing the health risk associated with consuming tilapia is to trim and cook the fish properly, that is, removing the viscera of tilapia can greatly reduce the amount of As ingested and consequently reduce the health risks. PMID- 12900945 TI - Biochemical and histopathological effects of glyphosate herbicide on Nile tilapia (Oreochromis niloticus). AB - In Oreochromis niloticus that had been exposed for 3 months to sublethal concentrations (5 and 15 ppm) of the commercial glyphosate herbicide (C(3)H(8)NO(5)P) Roundup, the organs exhibited varying degrees of histopathological change. In the gills filament cell proliferation, lamellar cell hyperplasia, lamellar fusion, epithelial lifting, and aneurysm were observed. In the liver there were vacuolation of hepatocytes and nuclear pyknosis. Kidney lesions consisted of dilation of Bowman's space and accumulation of hyaline droplets in the tubular epithelial cells. The structural damages could be correlated to the significant increase (p or = 15% change in the INR while receiving either COX-2 inhibitor, 4 showed this change for both agents. Adverse drug reactions were similar for each COX-2 inhibitor, but the rate of edema requiring medical intervention was higher in the rofecoxib group. Significant increases in the INR were observed in patients who were stable on warfarin therapy after the addition of therapy with rofecoxib or celecoxib. PMID- 12901033 TI - Oral bioavailability of pantoprazole suspended in sodium bicarbonate solution. AB - The bioavailability of pantoprazole when administered as a suspension in sodium bicarbonate solution and as the oral tablet was studied. In an open-label, randomized, two-period crossover study, healthy fasting subjects received either one enteric-coated 40-mg pantoprazole tablet by mouth with 240 mL of water or 20 mL of a suspension prepared from one crushed pantoprazole tablet and 840 mg of sodium bicarbonate solution and administered via a nasogastric tube. Treatments were separated by a 48-hour washout period. Blood samples were collected at intervals up to 24 hours after drug administration for measurement of pantoprazole concentration by high-performance liquid chromatography (HPLC) and estimation of pharmacokinetic values. A separate study was conducted to determine pantoprazole's stability in the suspension for up to three months at 25, 5, and 20 degrees C; concentrations were measured by HPLC. Twelve subjects completed the study. The suspension yielded pantoprazole Cmax values similar to those of the tablet formulation, but the drug was 25% less bioavailable. There was no lag time for the suspension. The suspension was stable for up to two weeks at 5 degrees C and up to three months at -20 degrees C. A suspension of pantoprazole in sodium bicarbonate solution yielded a Cmax similar to that of the tablet formulation, and the drug was quickly absorbed. However, bio-availability was slightly lower with the suspension than with the tablet. PMID- 12901034 TI - Evaluation of the appropriate use of albumin in adult and pediatric patients. AB - The appropriateness of albumin use and baseline albumin usage patterns were studied. Institutional practice patterns regarding the use of albumin were compared to criteria established by an independent expert panel. Fifty-three institutions, all of which were members of VHA or the University Health-System Consortium, participated in the evaluation. Investigators collected data over an eight-week period from the medical records, pharmacy records, and hospital billing data of adult (18 years of age or older) and pediatric (age 1-17 years) patients for whom albumin was prescribed. Data collected included patient specific information, the prescribing physician's specialty area, patient location (level of care) when albumin was prescribed, primary reasons for prescribing albumin, and details of albumin use. Data were collected for 1649 adult and 23 pediatric patients. Albumin was prescribed inappropriately in 57.8% and appropriately in 28.2% of adults; appropriateness of use was unknown in 14% of the patients reviewed. The most common indication for albumin use was hypotension/hypovolemia (23.9%), followed by bypass-pump priming (16.3%), intradialytic blood pressure support (9.6%), and serum albumin values less than 2 g/dL (8.6%). Albumin was prescribed inappropriately 100% of the time when used for intradialytic blood pressure support, low serum albumin values, and acute respiratory distress syndrome. The most appropriate use of albumin occurred in patients with postsurgical hypotension and hypovolemia (67.8%), nephrotic syndrome (79.3%), non-hemorrhagic shock (44.3%), hemorrhagic shock (51.9%), and cirrhosis and paracentesis (31.3%). Albumin was inappropriately prescribed for 57.8% of adult patients and 52.2% of pediatric patients. The mean number of total grams used by patients receiving albumin appropriately was similar to those patients inappropriately receiving albumin. PMID- 12901035 TI - Evaluation of a model for counseling patients with dyspepsia in Swedish community pharmacies. AB - The outcomes of a counseling model designed to help pharmacists care for customers seeking nonprescription treatment for dyspepsia in Swedish community pharmacies were studied. A protocol containing 10 key questions based on clinical guidelines about dyspepsia symptoms and on studies of drug-related problems (DRPs) was implemented in six community pharmacies in Sweden. During two weeks in April 2002, all adult customers seeking self-care for dyspepsia were asked to participate in the interview. Depending on a customer's answers, pharmacy personnel gave advice, intervened for DRPs, or referred the customer to a physician. Customers who received advice or intervention were followed up by telephone. A total of 319 customers who met the study criteria agreed to participate. Of these, 183 (57%) received self-care advice, 37 (12%) had DRPs, and 39 (12%) were referred to a physician because of their symptoms. Of the 198 customers fulfilling the criteria for a follow-up interview, 130 (66%) participated. Nearly all of these customers were satisfied with their visit to the pharmacy, and 9 in 10 were pleased with the advice received. Eighty-five percent claimed they had followed the self-care advice, and two in three said they felt better. DRPs were resolved in 19 (86%) of the 22 customers with DRPs who were followed up. Only one customer in five who was advised to see a physician actually did so. A counseling model designed to discover and resolve problems related to symptoms and drug use appeared to have a favorable impact on outcomes in customers with dyspepsia seeking nonprescription drug treatment in Swedish pharmacies. PMID- 12901036 TI - Assessment of potential digoxin-rabeprazole interaction after formulary conversion of proton-pump inhibitors. PMID- 12901038 TI - Pharmacists' use of and attitudes and beliefs about alternative medications. PMID- 12901037 TI - Applying patient selection criteria for drotrecogin alfa therapy in practice. PMID- 12901039 TI - A pharmacist-driven antimicrobial approval program at a Veterans Affairs hospital. PMID- 12901040 TI - ASHP guidelines on the pharmacist's role in immunization. PMID- 12901041 TI - Actions of the ASHP Board of Directors--meeting of April 14-16, 2003. PMID- 12901042 TI - Patient safety mandates: a means to an end? PMID- 12901043 TI - Additional laser benefits? PMID- 12901044 TI - Mouth gives "warning signs" of larger health problems. PMID- 12901045 TI - Supply of stem cells found in baby teeth. PMID- 12901046 TI - Saliva may help diagnose oral cancer. PMID- 12901048 TI - The high cost of low morale. PMID- 12901047 TI - Kids with braces need more than a toothbrush, study says. PMID- 12901049 TI - Maximizing the use of an intraoral camera. PMID- 12901050 TI - Laser-assisted amalgam removal with no local anesthetic. PMID- 12901051 TI - Full-mouth reconstruction. Restoring vertical dimension, speech, and self confidence. PMID- 12901052 TI - Rx for success: teamwork and trust. Creating a winning doctor/technician relationship. PMID- 12901053 TI - Combining technologies for single-visit treatment. PMID- 12901054 TI - Tooth surface loss. The least treated condition? PMID- 12901055 TI - Traumatic injuries of the teeth: current treatment modalities. AB - Teeth that have been traumatized and exhibit craze lines, fracture into enamel, fracture into dentin, root fracture, concussion, or subluxation should be pulp tested 3 months after the accident. If the tooth has a vital pulp, root canal treatment likely will not be required. Teeth that have a crown-root fracture should receive immediate endodontic therapy, and--if needed--can be extruded orthodontically. An intruded tooth should have endodontic therapy the same day, with calcium hydroxide placed into the canal. Teeth that have been extruded, displaced in any direction, or totally avulsed must have the canal completely cleaned, shaped, and filled with calcium hydroxide within 1 or 2 weeks post trauma. Clinical research concerning traumatized teeth is ongoing, and it is incumbent upon each practitioner to keep abreast of the current literature. This review has discussed treatment modalities that are recommended at this time. PMID- 12901056 TI - Success with occlusal splints. PMID- 12901057 TI - Automated external defibrillators, Part 2. Application. PMID- 12901058 TI - Clinical applications of appliance therapy in general dental practice, Part 3. Minor tooth movement for adults. PMID- 12901059 TI - Optimizing infant and toddler oral health. The importance of early intervention. PMID- 12901060 TI - A businesslike examination of overhead. Utilizing CPA-style tools to improve the DDS bottom line. PMID- 12901061 TI - Technology. It's all about relationships. PMID- 12901062 TI - Practical procedures & aesthetic dentistry. PMID- 12901063 TI - The wax injection method of replicating teeth. PMID- 12901064 TI - Three-unit fixed partial dentures versus single-tooth implant restorations. PMID- 12901065 TI - Magnification in endodontics: the use of the operating microscope. PMID- 12901066 TI - Creating the new patient experience--Part 1. PMID- 12901067 TI - Prevention of sports-related dental trauma: the role of mouthguards. PMID- 12901068 TI - Seeing is believing: the impact of the operating microscope on nonsurgical endodontic treatment. PMID- 12901069 TI - Evolving technologies for improved ergonomics: selecting an appropriate electric handpiece. PMID- 12901071 TI - Fiber-reinforced resin fixed partial dentures: a 3-year update. PMID- 12901070 TI - Adhesive reattachment of a tooth fragment: the biological restoration. AB - Recent developments in restorative materials, placement techniques, preparation design, and adhesive protocols allow clinicians to predictably restore fractured teeth. Using a minimally invasive approach, treatment of the maxillary anterior region can be effortlessly completed within a single appointment. If the original tooth fragment is retained following fracture, the natural tooth structures can be reattached using adhesive protocols to ensure reliable strength, durability, and aesthetics. This article discusses the adhesive reattachment of a tooth fragment to a fractured incisor using a conservative preparation technique. PMID- 12901072 TI - Reconnaissance soil geochemical survey of Gibraltar. AB - The extreme density of population of Gibraltar, situated at the southern tip of Spain, exerts considerable pressure on land use and thus future planning is of utmost importance. An initial reconnaissance soil geochemical survey of Gibraltar was based on 120 surface samples (0-15 cm) taken from a wide range of exposed, either bare soil or vegetated sites, to provide the optimum geographical distribution. The 'total' elemental concentrations of 26 elements (Li, Na, K, Be, Mg, Ca, Sr, Ba, Al, La, Ti, V, Cr, Mo, Mn, Fe, Co, Ni, Cu, Ag, Zn, Cd, Pb, P, S, As) were determined by nitric/percholric acid digestion followed by inductively coupled plasma-atomic emission spectrometry (ICP-AES) analysis. The reconnaissance data shows that the spatial distribution of various elements depended on previous and present land use. Most elements (Ca, Cr, Mg and Mn excluded) exhibited relatively high concentrations in civilian and natural soils. Trends have been established for many elements, and concentrations exceeding guideline values have been found in certain areas of Gibraltar. This reconnaissance of Gibraltar is at present being followed by a more detailed baseline geochemical survey, which will establish the extent and magnitude of the variations in major and trace elements in soils and dusts, assess the impact of industrial, commercial and urban development on the geochemical landscape and to make recommendations concerning sustainable development. PMID- 12901073 TI - Is there a relationship between soil and groundwater toxicity? AB - Part IIA of the Environmental Protection Act 1990 requires environmental regulators to assess the risk of contaminants leaching from soils into groundwater (DETR, 1999). This newly introduced legislation assumes a link between soil and groundwater chemistry, in which rainwater leaches contaminants from soil into the saturated zone. As the toxicity of both groundwater and overlying soils is dependent upon the chemicals present, their partitioning and their bioavailability, similar patterns of soil, leachates and groundwater toxicity should be observed at contaminated sites. Soil and groundwater samples were collected from different contaminated land sites in an urban area, and used to determine relationships between soil chemistry and toxicity, mobility of contaminants, and groundwater chemistry and toxicity. Soils were leached using water to mimic rainfall, and both the soils and leachates tested using bioassays. Soil bioassays were carried out using Eisenia fetida, whilst groundwater and leachates were tested using the Microtox test system and Daphnia magna 48 h acute tests. Analysis of the bioassay responses demonstrated that a number of the samples were toxic to test organisms, however, there were no significant statistical relationships between soil, groundwater and leachate toxicity. Nor were there significant correlations between soil, leachates and groundwater chemistry. PMID- 12901074 TI - Soil ingestion by sheep grazing the metal enriched floodplain soils of mid-Wales. AB - Floodplain soils within and downstream from the mineralised and mined areas of mid-Wales, are contaminated by metals, especially Pb, because of historical and contemporary fluvial pollution. Rates of soil ingestion by sheep grazing these sites have been quantified to establish the relative importance of the soil-plant animal and soil-animal pathway of metals. The highest rates of soil ingestion occurred during the winter/spring period. During March, soil ingestion exceeded 30% of the D.M. intake at 2 of the 11 sites investigated. The total daily intake of metals by sheep reflects the degree of soil metal enrichment, and is elevated during the winter/spring period, coincident with the higher rates of soil ingestion and the generally higher pasture herbage metal concentrations. Because the soil-plant transfer of Pb is low, ingested soil is often the major pathway of this metal to sheep. This is especially evident in March and May when on average 80.0 and 82.9%, respectively of the Pb intake was via soil ingestion. At one site in May, 97% of the Pb intake was attributable to ingested soil. Even when soil plant transfers are not so low, as found for Cu and Zn, ingested soil can occasionally supply greater than 60% of these metals to the animal. However, despite the potential importance of soil ingestion, little is known about the availability to and absorption of soil-borne metals by animals. PMID- 12901075 TI - Contaminant source apportionment by PIMMS lead isotope analysis and SEM-image analysis. AB - By combining scanning electron microscopy (SEM) image analysis and laser ablation plasma ionisation multi-collector mass spectrometry (LA-PIMMS), high precision lead isotope analyses can be obtained from individual metal-rich particles. Soils from Wolverhampton and Nottingham were sampled on the basis of high Pb concentrations or brownfield location. Pressed powder pellets of each were rastered by LA-PIMMS to obtain a bulk Pb-isotope signature. The results plot along an apparent mixing line between the major sources of lead contamination in the UK, that is UK ore deposits and alkyl-lead from petrol additives (Australian ore). Two particularly lead-rich soils were chosen to investigate the lead distribution and isotope variability between size and density fractions. The fine grained and low-density fractions contained most of the lead and have Pb-isotope ratios comparable with the bulk soils. By contrast, the small, lead-enriched denser fractions contained only a minor proportion of the total lead but Pb isotope signatures indicating relative enrichment in one or other of the end members from the mixing line. Further characterisation of individual Pb-rich grains is in progress. PMID- 12901076 TI - Predicting the risk of cobalt deprivation in grazing livestock from soil composition data. AB - The objective was to derive predictive equations for acetic acid-extractable cobalt (A-ECo) in soils so that extensive national databases for total (T) Co in soils and stream sediments could be converted to 'plant available' concentrations for the purpose of predicting risk of Co deficiency in grazing livestock. Data on the chemical and physical properties of 103 soils from 15 different parent materials and 54 soil series in England and Wales were used. Ranges for the mean values for parent materials were: TCo, 5.0-20.4 and A-ECo, 0.20-1.30 mg kg-1; percentage (P) A-ECo, 3.4-13.5; soil manganese (Mn) 268-1174 mg kgDM-1; pH, 3.7 8.0. There were significant effects of parent material on all parameters with Chalks, Old and New Red Sandstones particularly low in A-ECo. Multiple linear regression yielded the following equation for predicting A-ECo, which accounted for 56% of the variance with 12 outliers, including the lowest pH values, omitted: [figure: see text] REML was used on the complete, unbalanced, log transformed data set to fit a Generalised Mixed Model with parent material as random effect and soil Mn and pH as fixed effects; the effect of parent material was no longer significant. It was concluded that A-ECo can be satisfactorily predicted for most soils in England and Wales from TCo, TMn and soil pH. PMID- 12901077 TI - A comparison of metals in sediments and water in the River Nahr-Ibrahim, Lebanon: 1996 and 1999. AB - Rivers whose basins are underlain by carbonate rocks exhibit high pH, lower desorption of metals and possess high buffering capacity against acidic inputs to the river. The catchment of River Nahr-Ibrahim, Lebanon, is largely underlain by limestone. Compared to neighbouring countries, Lebanon is relatively fortunate since precipitation is high. However, recently a warming in temperature and a drop in precipitation has occurred, thus causing low water levels in rivers. The objective of this study is to investigate the variation of the total metal content (Fe, Mn, Zn, Cu, Pb and Cd) in bed sediments and water of River Nahr Ibrahim between 1996 and 1999 (two years); and relate these variations to the effect of changes in human activities and/or due to the variations of precipitation rate, temperature and pH of water. Bed load sediments and stream water were collected simultaneously from five sampling sites. Water pH and temperature were determined in situ. Sediment samples were dried at room temperature and sieved; the sediment size < 75 microns size was retained. Water was analysed for major constituents and trace metals. Metals were extracted from sediments with aqua regia. Metal concentration in water and sediments were determined using ICP-MS technique. Data revealed a drop in metal concentrations (Zn, Cu, Pb, Cd) in sediments at quarry site after its closure. The decrease in precipitation rate, lowering the level of water and the dilution of industrial discharges and decrease in water pH led most probably to the desorption of metals from sediments into the water. PMID- 12901078 TI - Sorption of Cu and Zn by bone charcoal. AB - Bone charcoal is being developed as a treatment for decontaminating polluted water. In particular, its potential to adsorb metal species from contaminated water supplies is being examined. In this study, bone charcoal was used in batch and column experiments designed to investigate the sorption of two cationic metals Cu and Zn. The data from batch experiments fitted the Langmuir isotherm and so theoretical maximum binding capacities could be obtained. These values were compared with experimental values derived from the column experiments. The mechanisms involved in the immobilisation were also investigated. PMID- 12901079 TI - Application of a water quality model in the White Cart water catchment, Glasgow, UK. AB - Water quality models of urban systems have previously focused on point source (sewerage system) inputs. Little attention has been given to diffuse inputs and research into diffuse pollution has been largely confined to agriculture sources. This paper reports on new research that is aimed at integrating diffuse inputs into an urban water quality model. An integrated model is introduced that is made up of four modules: hydrology, contaminant point sources, nutrient cycling and leaching. The hydrology module, T&T consists of a TOPMODEL (a TOPography-based hydrological MODEL), which simulates runoff from pervious areas and a two-tank model, which simulates runoff from impervious urban areas. Linked into the two tank model, the contaminant point source module simulates the overflow from the sewerage system in heavy rain. The widely known SOILN (SOIL Nitrate model) is the basis of nitrogen cycle module. Finally, the leaching module consists of two functions: the production function and the transfer function. The production function is based on SLIM (Solute Leaching Intermediate Model) while the transfer function is based on the 'flushing hypothesis' which postulates a relationship between contaminant concentrations in the receiving water course and the extent to which the catchment is saturated. This paper outlines the modelling methodology and the model structures that have been developed. An application of this model in the White Cart catchment (Glasgow) is also included. PMID- 12901080 TI - Sorption of the natural endocrine disruptors, oestrone and 17 beta-oestradiol in the aquatic environment. AB - Adsorption of hydrophobic contaminants at the particle/water interface is one of the key processes controlling their fate in the aquatic environment. The sorption of the natural female hormones oestrone and 17 beta-oestradiol has been studied under simulated riverine conditions. Both the kinetics and the effects of varying fundamental environmental parameters (e.g. sediment properties) on the thermodynamic equilibrium partition coefficient (Kp) have been studied in continuous and batch sorption experiments, respectively. Results showed that the sorption of oestrone and 17 beta-oestradiol by sediment was relatively slow, reaching equilibrium in 50 days. In addition, relatively small adsorption of both oestrone and 17 beta-oestradiol onto the sediment was observed, with Kp values between 200 and 250 mL g-1. The comparable Kp values of the two compounds reflect their structural similarity. It can be concluded that the two endocrine disruptors, oestrone and 17 beta-oestradiol remain primarily in association with the aqueous phase. PMID- 12901081 TI - The application of solid-phase micro-extraction (SPME) to the analysis of polycyclic aromatic hydrocarbons (PAHs). AB - There are many established extraction techniques regularly used in the isolation and analysis of PAHs and similar organic compounds from various phases. These include Soxhlet or ultrasonic extractions from solids, and liquid-liquid or solid phase extraction from aqueous samples. However, these methods have some inherent disadvantages; most require large volumes of organic solvents, they can be time consuming and many involve multi-step processes that always present the risk of the loss of some analytes (Zhang et al., 1994). Solid-phase micro-extraction (SPME) is a relatively new technique that has been used with much success in the analysis of a variety of compounds including PAHs. Experiments are being carried out to determine the optimum range of conditions for the extraction of a range of PAHs. Parameters under investigation include temperature, equilibration time, salinity and compound concentration. Presented here are some preliminary experiments into the applicability of SPME for PAH analysis. Further work will investigate the reproducibility of the technique, limits of detection and matrix effects. When an optimised method has been developed the technique will be used in investigations into PAH profiles in sediment cores. PMID- 12901083 TI - The relationship between epilithic periphyton (biofilm) bound metals and metals bound to sediments in freshwater systems. AB - Surficial sediments and epilithic periphyton (biofilm) were sampled from six sites on the River Churnet and five sites on the River Manifold in Staffordshire and analysed for cadmium (Cd), copper (Cu), lead (Pb) and zinc (Zn). The sites demonstrated a wide range of sediment trace metal concentrations determined by inductively coupled plasma optical emission spectrometry (ICP-OES) and atomic absorbtion spectroscopy (AAS). Biofilm was removed from the substrate using physical abrasion and 0.005 M ethylenediaminoethanetetra-acetic acid (EDTA) extractant. The European Standards, Measurements and Testing Programmes (BCR) operationally defined geochemical speciation scheme was used to determine the exchangeable, acid soluble fraction of the sediments. Significant positive correlations were determined between the EDTA extractable biofilm and the exchangeable sediment fraction for Cd, Cu and Zn but not for Pb. Natural epilithic periphyton may be a potential metal biomonitor particularly of Cu, Cd and Zn in aquatic systems and provide supporting information in relation to potential sediment toxicity. PMID- 12901082 TI - Seasonal variation of total dissolved arsenic and arsenic speciation in a polluted surface waterway. AB - Seasonal differences in the dissolved arsenic concentration and speciation in a contaminated urban waterway in northwest England have been determined using a coupled ion chromatography-inductively coupled plasma-mass spectrometry (IC-ICP MS) technique. Waters sampled in the vicinity of an industrial works during relatively dry conditions in April 2000 were found to contain total arsenic concentrations (sigma As) of up to 132 micrograms L-1, more than an order magnitude greater than the 4 micrograms L-1 maximum found in December 2000. The difference in sigma As between the April and December sampling periods is speculated to be largely due to the irregular anthropogenic supply of arsenic to the watercourse. For both sampling periods, the dissolved arsenic was exclusively inorganic in nature and had an As(V)/sigma As ratio of between 0.6 and 0.8. Analysis of samples taken downstream of the industrial site, after the confluence with a relatively As-poor stream, revealed that As(III), As(V) and sigma As concentrations were lower than would be expected from conservative mixing. The As(V)/sigma As ratio was also observed to decrease markedly. The loss of arsenic from solution is thought to be due to adsorption on the iron oxyhydroxide-rich sediment observed to coat the riverbed downstream of the confluence. The reduction in the As(V)/sigma As ratio is believed to be due to the more rapid adsorption of As(V) compared to that of As(III). Deviations from conservative behaviour were more marked during the relatively dry April 2000 sampling period and suggest the increased importance of adsorption processes controlling arsenic availability during this time. PMID- 12901084 TI - Metal contamination of active stream sediments in upper Weardale, northern Pennine Orefield, UK. AB - In the Upper Weardale area the headwaters of the River Wear bisect the Northern Pennine Orefield, where Pb-Zn-F-Ba vein-type mineralisation has been exploited since the Roman Conquest. The area contains evidence of open pit, underground and hydraulic mining of base metal ores, associated mineral processing and smelting, exploitation of ironstones during the industrial revolution, recent extraction of fluorite and active quarrying. The aim of this study was to determine the extent of modern sediment contamination arising from these past activities. Samples of active stream sediments were collected from all major drainage channels at 1 km intervals. The sediments were analysed for Pb, Zn, Ba, Mn, Fe, Co, Ni, Cu, Cr, As, Sb, Ag and compared to data from earlier regional geochemical surveys of low order drainage samples using ArcView software. The significance of contamination levels was assessed using the Ontario aquatic sediment quality guidelines. Our results indicate widespread contamination of some major drainages by Pb, Mn, Zn and As at concentration levels anticipated to significantly affect use of the sediments by benthic organisms. Furthermore, Pb contamination shows persistence in stream sediments downstream towards agricultural areas of the floodplain and drinking water abstraction points, above which interaction with colliery mine water discharges may occur. PMID- 12901085 TI - Nitrate levels and the age of groundwater from the Upper Devonian sandstone aquifer in Fife, Scotland. AB - The tritium concentrations in 13 groundwater samples from boreholes throughout the Upper Devonian sandstone aquifer of Fife have been measured. Due to atmospheric variations in tritium concentrations over the last century, this radioactive tracer can be used as a groundwater age indicator. In this study, the groundwater tritium concentrations have allowed for the area to be divided into three zones, and the variable chemistry of the groundwater samples, including the problem of recent elevated nitrate levels in the Fife Aquifer, has been interpreted in terms of their relative ages. PMID- 12901086 TI - Sediment and porewater geochemistry in a metal contaminated estuary, Dulas Bay, Anglesey. AB - Remobilisation of contaminant metals from sediments can occur by chemical, biological or physical changes. This in turn can lead to contaminant fluxes to the porewaters and ultimately the water column. The aim of the research presented here is to document post-depositional controls on metal mobility and fluxes in a heavily metal-contaminated estuary. This will allow for an improved understanding of the impact of contaminated sediments on water quality from both a short-term and long-term perspective. Dulas Bay is situated on the east coast of Anglesey, North Wales, and receives polluted waters from Parys Mountain. Metals within Dulas Bay sediments show surface enrichment and variations in mineralogical form. Diagenesis clearly plays a role in post-depositional behaviour of the metals, forming sulphides and potentially carbonates. The presence of a dominant exchangeable/carbonate fraction, and elevated porewater metals, in this sulphidic system is significant and could indicate the presence of freshwater diagenesis, or, reflect the high levels of metals in the sediment. PMID- 12901087 TI - The effect of diesel fuel on common vetch (Vicia sativa L.) plants. AB - When petroleum hydrocarbons contaminate soil, the carbon:nitrogen (C:N) ratio of the soil is altered. The added carbon stimulates microbial numbers but causes an imbalance in the C:N ratio which may result in immobilization of soil nitrogen by the microbial biomass, leaving none available for plant growth. As members of Leguminosae fix atmospheric nitrogen to produce their own nitrogen for growth, they may prove more successful at growing on petroleum hydrocarbon contaminated sites. During a wider study on phytoremediation of diesel fuel contaminated soil, particular attention was given to the performance of legumes versus other plant species. During harvesting of pot experiments containing leguminous plants, a recurring difference in the number and formation of root nodules present on control and contaminated Common vetch (Vicia sativa L.) plants was observed. The total number of nodules per plant was significant reduced in contaminated plants compared to control plants but nodules on contaminated plants were more developed than corresponding nodules on control plants. Plant performance of Common vetch and Westerwold's ryegrass (Lolium multiflorum L.) was compared to illustrate any difference between the ability of legumes and grasses to grow on diesel fuel contaminated soil. Common vetch was less affected by diesel fuel and performed better in low levels of diesel fuel contaminated soil than Westerwold's ryegrass. The total amount of diesel fuel remaining after 4 months in Common vetch planted soil was slightly less than in Westerwold's ryegrass planted soil. PMID- 12901088 TI - Preliminary evidence for the metabolism of benzo(a) pyrene by Plantago lanceolata. AB - Polycyclic aromatic hydrocarbons (PAHs) have long been recognised as potential carcinogens in animals in which biotransformation into reactive metabolites can lead to DNA damage. In animals PAHs metabolism mainly occurs in hepatic microsomes and is associated with the cytochrome p-450 mediated mixed functional oxidase (MFO) system. PAH metabolism in plants has been shown to occur via a similar enzyme system, but has received relatively little attention. This study is looking at how the plant species Plantago lanceolata metabolizes benzo(a)pyrene (B(a)P), which is one of the PAHs whose metabolism has been studied extensively in animals. The aim of the work is to establish firstly that the B(a)P is taken up and secondly that it is biotransformed by the plant to products possibly similar to those found in animals. This work is achieved by using C-14-B(a)P along with whole body autoradiography, scintillation analysis and chromatography techniques to locate the B(a)P and its metabolites. PMID- 12901089 TI - A preliminary study of the phycological degradation of natural stone masonry. AB - For many years it has been realised that the weathering of stone is not merely determined by physical and chemical factors but also by biological agents. When the stone in question is a historic building or monument, the damage done constitutes an irretrievable loss of our heritage and history. Laboratory studies have commenced in Paisley to study the effect of photoautotrophs on the major sedimentary rock forming minerals, with a view to expanding this work to study the overall effect of these micro-organisms on heritage masonry. Tests were carried out on Albite, Calcite, Dolomite, Orthoclase, Siderite and Quartz, using axenic cultures of the following: Chlorella vulgaris, Chlorococcum tetrasporum, Scenedesmus obliquus, Oocystis marsonii, Stichococcus bacillaris. The rock chips were immersed in either water or bolds basal media and exposed to a mix of the micro-organisms listed above and then tested weekly for their pH, fortnightly for the waters chemical composition using inductively coupled plasma-atomic emission spectrometry (ICP-AES) and visually utilising the university's SEM facilities. Work so far has revealed biologically mediated etching of minerals, a well defined pH profile over a period of 90 days, as well as a variety of elemental release patterns for the different minerals. PMID- 12901090 TI - Mechanisms of copper tolerance by Armeria maritima in Dolfrwyong Bog, north Wales -initial studies. AB - Preliminary field studies were carried out at Dolfrwynog Bog in July 2000. Replicate samples of water, Armeria maritima plants and the soils adhering to its roots were collected and analysed for copper. Concentrations of up to 6486 mg kg 1 of copper in the soils were recorded. Accumulation of copper by the plant as expressed by concentration factors (CF) show that it is acting mainly as a copper excluder. Of the copper that is taken up, most of it is retained within the roots with very little being transported to the shoots of the plant. Moreover, a further possible mechanism of tolerance is exhibited by the excretion of copper through its decaying leaves. Towards the use of in vitro cultures to study the copper tolerance mechanisms in A. maritima a micropropagation protocol has been developed. The ex vitro plants have been rooted and established in compost. PMID- 12901091 TI - A pilot study of the relationship between multiple sclerosis and the physical environment in northwest Ireland. AB - Multiple sclerosis (MS) is a neurological disease of unknown origin, but it is though likely to have an environmental component in addition to genetic predisposition. It appears common in areas with underlying radon-producing granites. Radon exposure before the age of 15 years is postulated as a contributory cause in genetically susceptible individuals. Pilot studies were undertaken, with results which indicate that radon exposure may be a factor, and that the hypothesis warrants further investigation. PMID- 12901092 TI - The illusion of environmental iodine deficiency. AB - The distribution of endemic goitre in England and Wales was compared with the distribution of environmental iodine (atmospheric deposition, soil, surface water). Despite a very clear goitre belt through the west of England and Wales there was no patterning in the environmental iodine distribution. A clear seasonal variation in depositional iodine exists, with an unusually high concentration of iodine in March 1997. The temporal variation in iodine concentration is determined at the monthly and not the annual level. The presence of endemic goitre is no indicator of how iodine is distributed in the environment or vice versa! PMID- 12901093 TI - [Role of analytical chemistry in the development of new drugs]. AB - In this paper, the role of analytical chemistry in drug research, its development and some basic research related to developing new drugs are concisely discussed, including the establishment of quality control standards, finger prints of natural medicine products, resolution of chiral compounds and drug metabolism studies. The incorrect point of view of some people on pharmaceutical analysis is mentioned and the indispensability of this discipline in drug research and development is also emphasized. PMID- 12901094 TI - [Systemic breeding of the new variety of Biantiao ginseng (Panax ginseng)]. AB - OBJECTIVE: To breed new varieties of Biantiao ginseng for high yield and fine quality. METHODS: Systemic breeding methods were applied. About 3,000 outstanding Biantiao ginseng roots were selected and planted in breeding field, and self crossed for four generations. During the course, inferior lines or plants were rejected. Then strain comparison, identification of resistance to black-speck disease, and analysis of active compositions were carried out. RESULTS: "Biantiao 1" (BT1), the first new variety of Biantiao ginseng, with green stems and thick, long, elegant roots and median resistance to black-speck disease, has been harvested since 20 years. The percentage of Biantiao ginseng roots and yield were 15% and 30% higher than the control's respectively. The content of total ginsenosides and the main monomers was 1.8%-2.5% higher than the control's. The characteristics of overground part and root of BT1 were uniform and stable. CONCLUSIONS: BT1, a new excellent ginseng variety, has a good potential value to be generalized in ginseng production. PMID- 12901095 TI - [Effects of fungus on the growth of Dendrobium candidum and D. nobile in vitro culture]. AB - OBJECTIVE: To examine effects of fungus AR-18 (Epulorhiza sp.) on the growth of Dendrobium candidum and D. nobile in vitro culture. METHODS: Effects of fungus AR 18 on fresh weight and dry weight of D. candidum and D. nobile were studied in vitro culture, when agar was used as rest, and effects of fungus AR-18 and nutrients on fresh weight and dry weight of D. candidum and D. nobile were studied in vitro culture, when vermiculite was used as rest. RESULTS: In agar medium, effects of fungus AR-18 on fresh weight and dry weight of D. candidum and D. nobile were not significant (P > 0.05). However, in vermiculite medium, the effect of fungus AR-18 on fresh weight of D. nobile was significant (P < 0.05). Fresh weight of D. nobile inoculated with fungus AR-18 was increased by 16% compared with the non-inoculated ones (control). The effect of fungus AR-18 on dry weight of D. nobile was very significant (P < 0.01). Dry weight of D. nobile inoculated with fungus AR-18 was increased by 21% compared with the control. In vermiculite medium, the effect of combination treatment (Fungus AR-18 + Distilled H2O) on fresh weight of D. nobile was very significant (P < 0.01). Fresh weight of D. nobile inoculated with fungus AR-18 was increased by 47% compared with the control, while the effect of combination treatment (Fungus AR-18 + Nutrients) on fresh weight of D. nobile was not significant (P > 0.05). In addition, the effect of combinations treatment (Fungus AR-18 + Nutrients) and (Fungus AR-18 + Distilled H2O) on dry weight of D. nobile was not significant (P > 0.05). So the combination treatment (Fungus AR-18 + Distilled H2O) was the best for the growth of D. nobile. In vermiculite medium, effects of fungus AR-18 and nutrients on fresh weight and dry weight of D. candidum were not significant (P > 0.05). CONCLUSION: It is important in vitro culture to select a proper rest for the control of the growth of fungus to establish a beneficial symbiotic relationship between plants and fungi. If the fungus AR-18 is utilized as biological manure in got-up planting of D. nobile, a good result may be obtained. PMID- 12901096 TI - [Effects of growth regulator, full exposure and mist spray on the cutting of Eucommia ulmoides]. AB - OBJECTIVE: To study the effects of growth regulator on the cutting method of Eucommia ulmoides seedlings. METHODS: Treated with H2O (CK), NAA, 6-BA and NAA + 6-BA for 4 hours, respectively, cuttings of 4-year and 20-year trees were cultured for 45-60 days under the shed or full exposure and mist spray. The growth of the cuttings was observed periodically. The number of the roots, the root length, the callus and the death rate were investigated after harvest. RESULTS: Compared with the cutting from 4-year-old tree, the cutting from 20-year old tree developed better. The rate of stroking root in treated group was 14 times as much as CK group. Other agronomic traits such as the number of the roots, the root length, the callus, the number of the leaves and the death rate of the cuttings in treated group were all better than that of the cuttings in CK group. The cultural time in treated group reduced by 15 days than that of CK group. CONCLUSIONS: The present study shows that both full exposure mist spray and growth regulators have remarkable effects on cuttings of tree with different ages for rooting or forming callus. PMID- 12901097 TI - [Effects of endophytic fungal hyphae and their metabolites on the growth of Dendrobium candidum and Anoectochilus roxburghii]. AB - OBJECTIVE: To study the effects of hyphae and their metabolites of Mycena anoectochila, M. dendrobii and M. orchidicola on promoting the protocorns multiplication of D. candidum and platelet growth of A.roxburghii. METHODS: Add 20% (V/V) ground hypha or 10 mg/L ethyl acetate extracts of fermentation liquid respectively to the plant culture media. RESULTS: The ground hypha of 3 Mycena and ethyl acetate extracts of M. orchidicola could promote the protocorn multiplication rate of D. candidum. The ground hypha of M. dendrobii could increase the numbers of bud and improve the platelet growth of A.roxburghii. The ethyl acetate extracts of fermentation liquid of M. anoectochila increased the numbers of bud and M. orchidicola significantly improved the platelet growth of A. roxburghii. CONCLUSION: The effects of 3 Mycena Endophytic fungi on promoting growth of Dendrobium candidum and Anoectochilus roxburghii are related with their metabolites in and out of hypha. PMID- 12901098 TI - [Comparison and correlative analysis on characters of Rehmannia glutinosa Libosch. Varieties]. AB - OBJECTIVE: To compare the economical characters, yield characters and content of catalpol on Rehmannia glutinosa Libosch. varieties. METHODS: To study characters by field randomized block test and analysis of variance. To analyse the content of catalpol by HPLC. RESULTS: The results by analysis of variance were that the F value in plant width was 15.4 (F0.01 = 5.54), the F value in length of leaves was 12.2, the F value in width of leaves was 13.35, the F value in yield of single plant was 55.7 and the F value in content of catalpol was 8.03. The results by correlative analysis were that the linear correlation coefficient of signal plant yield with length of leaves was 0.9639, with width of leaves was 0.9073, with amount of earthnuts was 0.7060 and with plant fresh weight was 0.9950. The linear correlation coefficient of content of catalpol with plant width was 0.9169, with length of leaves was 0.7046 and with width of leaves was 0.7159. CONCLUSION: There were significant differences in plant width, length of leaves width of leaves, and yield of single plant and content of catalpol of Rehmannia glutinosa Libnosch. varieties. There were significantly positive correlations in signal plant yield with plant fresh weight, length of leaves, width of leaves and amount of earthnuts. There were significantly positive correlations in content of catalpol with plant width, length of leaves and width of leaves. PMID- 12901099 TI - [Antitumor effect of lidamycin-containing monoclonal antibody immunoconjugate with downsized-molecule]. AB - OBJECTIVE: To develop a novel monoclonal antibody (mAb) immunoconjugate with downsized-molecule and highly potent antitumor effects. METHODS: The mAb conjugate was prepared by linking lidamycin (LDM), an antitumor antibiotic with extremely potent cytotoxicity, to mAb Fab' fragment. The molecular weight of Fab' LDM was determined by non-reduced SDS-PAGE. The immunoreactivity of Fab'-LDM conjugate with cancer cells was detected by ELISA. The antitumor activity of the conjugate was determined by MTT assay, clonogenic assay, and animal model of transplantable colon carcinoma 26' (C26) in mice. RESULTS: The relative molecular mass of Fab'-LDM conjugate was approximately 65,000, suggesting a molar ratio of 1:1 between Fab' fragment and LDM in the conjugate. Fab'-LDM was reactive with hepatoma BEL-7402 and colon carcinoma 26 cells, but not reactive with KB cells. The cytotoxicity of Fab'-LDM conjugate to BEL-7402 cells, the antigen relevant cancer cells, was 13-fold more potent than that of free LDM, while the cytotoxicity of Fab'-LDM conjugate against C26 cells was 5.5-fold more potent than that of free LDM. However, the cytotoxicity of Fab'-LDM conjugate to KB cells, the antigen irrelevant cells, was similar to that of free LDM. Given by 3 intravenous injections, on day 1, 4 and 7, Fab'-LDM conjugate at doses of 0.05 mg/kg, 0.1 mg/kg and 0.2 mg/kg markedly suppressed the growth of colon carcinoma 26 in mice by 80%, 92% and 94%, respectively, whereas free LDM at 0.1 mg/kg suppressed the growth by 77%. The survival time of tumor-bearing mice was also increased by Fab'-LDM conjugate treatment. Fab'-LDM conjugate was more effective than equivalent unconjugated LDM (P < 0.01). CONCLUSIONS: The immunoconjugate composed of LDM and Fab' fragment that is characterized by downsized-molecule shows remarkable effectiveness against tumor growth. PMID- 12901100 TI - [Sequence analysis of the late region of human papillomavirus type 6 genome]. AB - OBJECTIVE: To study variations of genome late region of human papillomavirus type 6 (HPV-6) isolated from Chinese patients with condyloma acuminatum. METHODS: Using overlap PCR design, major capsid protein (L1) and minor capsid protein (L2) genes were separately amplified from clinical samples following HPV type determination, and were further assembled into HPV-6 genome late region sequences after inserting into plasmid and sequencing. RESULTS: Two sequences (GenBank accession number AY015006, AY015008) of HPV-6 late region were assembled, which are 2,869 bp long covering 35% HPV-6 genome and with complete open reading frames (ORFs) for L1 and L2. Compared with prototype sequence, nine point mutations were found, including four missense mutations, three of which were located in L2 ORF. Phylogenetic analysis indicated that the cloned sequences are classified into HPV 6b. CONCLUSIONS: HPV-6 genome late region, especially in L1 ORF, is very conserved (variation rate < 0.28%). The mutation from A to G at position 7081 in HPV-6 genome and from G to A at 7099 may represent region characteristics. This is the first report that describes sequence variations among genome late region of HPV-6 isolated from Chinese patients. PMID- 12901101 TI - [High level expression of human atrial natriuretic peptide in fusion form in E. coli system]. AB - OBJECTIVE: To construct recombinant plasmid with human atrial natriuretic peptide (ANP) gene in fusion form for stable and high level expression of genetic engineering product ANP in E. coli system. METHODS: Plasmids with ANP fusion gene were constructed by PCR and sub-cloning and fusion protein was expressed in E. coli system. High level expression of the fusion genes was enhanced by linking operons in tandem. ANP was released from the fusion protein with thrombin treatment and purified by chromatography. Genetic-engineered ANP was evaluated with the drug production requirement, and produced ANP was tested for its effects of lowering blood pressure and diuresis in vivo and vasodilation in vitro. RESULTS: A series of 4 plasmid pCW111-114, containing 1 to 4 gene operons respectively, were constructed, and the yields of fusion protein were 46%, 54.8%, 56.1% and 60.1% of total cell protein. Fusion protein in the form of inclusion body was isolated and purified, and then treated with thrombin to release ANP. After purification using chromatography columns, at least 3 mg/L culture of ANP with the purity higher than 96% was produced in shaking flask. Primary pharmacological evaluation showed the produced ANP had the effects of blood pressure lowering in vivo and diuresis in vivo and vasodilation in vitro, which was similar to the activity of standard ANP. CONCLUSIONS: By the protocol of fusion gene cloning and expression, the human ANP was produced successfully in E. coli system. PMID- 12901102 TI - [Fragile X mental retardation protein interacts with human NDK/Nm23-H2]. AB - OBJECTIVE: To investigate the physiological role of fragile X mental retardation protein (FMRP) and screen the proteins interacting with FMRP in human fetal hippocampus cDNA library. METHODS: Human fetal hippocampus cDNA library was constructed in yeast two-hybrid DAD vector pGAD10. Quality of the library was measured by picking up random colonies as templates for PCR testing. Proteins interacting with FMRP were screened by yeast two-hybrid system. Furthermore, the interaction site of FMRP was mapped in yeast. RESULTS: The average length of inserts of the two-hybrid library was 1.5 kb, and the ratio of recombinant colonies was about 90%. Human NDK/Nm23-H2 was found interacting with FMRP. NDK/Nm23-H2 interacted with FMRP exon 1-12, as well as FMRP isoforms without exon 12, and exons 14-17. NDK/Nm23-H2 couldn't interact with FMRP exon 1-6 and exon 2 7 fragments. CONCLUSIONS: Human NDK/Nm23-H2 can bind FMRP directly. The interaction site of FMRP is located at its exon 1-11. This interaction in vitro might alter the intracellular distribution of NDK/Nm23-H2, and even regulates the transcription and expression of FMRP. PMID- 12901103 TI - [Study on high-efficiency gene transfer of pseudotyped HIV vector]. AB - OBJECTIVE: To investigate the high-efficiency of pseudotyped HIV as gene transfer vector. METHODS: Three plasmids of pseudotyped HIV gene transfer vector system were transferred into packaging cell line 293T by Ca3 (PO4)2 precipitation method. GFP (Green Fluorescence Protein) or HSV-tk gene was constructed in the plasmid pHR'CS respectively (pHR'CS.GFP or pHR'CS.HSV-tk). The pseudotyped HIV particles were observed through electronic microscopy and were measured through spectrofluorophotometer. High titer pseudotyped HIV was harvested from volume of virus-producing cell supernatant and concentrated. Ovarian epithelial cancer cell line SKOV3 and normal human gingival fibroblast cell GF were infected by pseudotyped HIV. PCR and RT-PCR were resorted to demonstrate the successful transduction and transcription of the HSV-tk gene. After administration of GCV, the changes of those cells and apoptosis were observed through optical microscopy. The cytotoxicity efficacy of HSV-tk/GCV system was evaluated by MTT method. The growth inhibition rate (GIR) of cells and inhibition concentration 50 (IC50) were counted. RESULTS: The above plasmids were effectively transferred into 293T cell. A lot of pseudotyped HIV particles were observed through electronic microscopy. The virus supernatant had a high absorbing value at 510 nm through spectrofluorophotometer, which proved the existence of virus. After pseudotyped HIV infection, SKOV3 and GF had remarkable infection rate. 600 bp strand was seen through PCR and RT-PCR. Changes and apoptosis of cells followed by administration of GCV were observed. The MTT method showed that the cytotoxicity efficacy of GCV was high to SKOV3 and GF cell. CONCLUSIONS: The pseudotyped HIV is a high-efficiency gene transfer vector. PMID- 12901104 TI - [Propagation of hepatitis E virus in several cell lines including human embryo lung diploid cell KMB17]. AB - OBJECTIVE: To investigate the hepatitis E virus (HEV) sensitive cells and its tissue culture conditions. METHODS: The HEV from dejecta supernatant of patients with acute hepatitis E was amplified and activated by passaged in Rhesus. Then, the positive dejecta samples of infected monkeys were dealt with super centrifugation and virus for culture was obtained. Various human-derived (including KMB17, A549, BEL7402, and Hela) and non-human primates derived cells (Vero) were inoculated with HEV. Sensitivity of cells to HEV was measured by CPE (cytopathic effect), RT-PCR and immunofluorescence. RESULTS: CPE in KMB17, A549 and BEL7402 cells appeared during 7-9 days, meanwhile, cells shelled during 11-13 days on the first filial generation. The existence of HEV genome +RNA and replicated -RNA was still detectable by RT-PCR after the tenth filial generation. Neither CPE nor amplification of HEV genome RNA could be detected in Hela and Vero cells after the second to fourth filial generation. HEV could also be detected from inoculated KMB17 cells by immunofluorescence and RT-PCR. CONCLUSIONS: It indicates that KMB17, A549 and BEL7402 cells are sensitive to HEV under the experimental culture conditions, while Hela and Vero cells are insensitive. Tissue culture system of HEV in certain filial generation is established. PMID- 12901105 TI - [Significance of the unbalanced expression of Th1/Th2 type cytokines in human glioma]. AB - OBJECTIVE: To study the significance of the unbalanced expression of Th1/Th2 type cytokines in human glioma. METHODS: The gene expressions of Th1/Th2 type cytokines in 62 specimens of human glioma tissues, 4 glioma cell lines, peripheral blood mononuclear cell (PBMC) of 15 glioma patients, 5 specimens of normal adult brain tissue and 5 brain meningioma tissues were detected by semiquantitative reverse transcription polymerase chain reaction. IFN-gamma and IL-2 represent Th1 type cytokines. IL-4, IL-6, IL-10 and IL-13 represent Th2 type cytokines. RESULTS: There were obviously predominant expression of Th2 type cytokines in glioma cell lines (P < 0.01) and specimens of human glioma tissues (P < 0.01). The tendency of distinct expression of Th2 type cytokines in PBMC was also existent. There wasn't obvious discrepancy of the expression of two type cytokines in normal adult brain tissues and meningioma tissues. CONCLUSIONS: It is likely that the switching of Th1/Th2 type cytokines in gliomas as predominant expression of Th2 type cytokine genes is related to the origination of gliomas and the evasion of glioma cells from immune surveillance. PMID- 12901106 TI - [Influence of adjustment of balance of Th1/Th2 type cytokines on proliferation of glioma cells]. AB - OBJECTIVE: To study the influence of adjustment of balance of Th1/Th2 by external cytokines on proliferation of glioma cells. METHODS: The gene expressions of Th1/Th2 type cytokines in C6, 9L, U251 and SHG44 glioma cells were detected by semiquantitative reverse transcription polymerase chain reaction (RT-PCR). After the cells were induced with IFN-gamma + IL-4 McAb and IL-4 + IFN-gamma McAb respectively, we isolated the total RNA to proceed RT-PCR again. The evaluation of cell proliferation was proceeded by MTT assay method. RESULTS: There was obviously predominant expression of Th2 type cytokines in glioma cell lines (P < 0.01). The expression intensity of IFN-gamma was improved in IFN-gamma + IL-4 McAb groups and Th2 type cytokines were enhanced in IL-4 + IFN-gamma McAb groups. IFN-gamma and IL-4 McAb could cause the switch from Th2 to Th1, and could remarkably inhibit the proliferation of glioma cells in a dose-dependent way (P < 0.01). On the other hand, IL-4 and IFN-gamma McAb could strengthen the switch of Th2, and might stimulate the glioma cell growth, also in a dose-dependent way (P < 0.01). CONCLUSIONS: There is a Th2 preponderance in glioma cells. IFN-gamma and IL-4 McAb could regulate the switch from Th2 to Th0 or Th1, and inhibit the proliferation of glioma cells. PMID- 12901107 TI - [Construction and expression of huGM-CSF (9-127)-IL-6(29-184) fusion protein gene]. AB - OBJECTIVE: To construct and express huGM-CSF(9-127)-IL-6(29-184) fusion protein with high purity and both huGM-CSF and huIL-6 biologic activities. METHODS: The novel gene coding for the fusion protein of huGM-CSF(9-127)-IL-6(29-184) was constructed by strategy of step by step cloning in pBV220 expression vector. The amino acids 1-8 of huGM-CSF and the amino acids 1-28 of huIL-6 were deleted by PCR technique. The mutant huGM-CSF (9-127) and huIL-6 (29-184) cDNAs were linked via a linker sequence coding 15 amino acid residues (G-G-S-G-S)3. Fusion protein was expressed in E.coli host strain DH5 alpha. To obtain the fusion protein, Q Sepharose H.P. ion exchange chromatography and Sephacryl S-200 gel filtration were performed. The biologic activities were detected by MTT method. RESULTS: Fusion protein was expressed in E.coli host strain DH5 alpha in the form of inclusion body. The expression level was more than 25% of the total cell lysate. Through Q Sepharose H.P. ion exchange chromatography and Sephacryl S-200 gel filtration, huGM-CSF(9-127)-IL-6(29-184) fusion protein with high purity was obtained. The protein showed both huGM-CSF and huIL-6 biologic activities. The specific activity of huGM-CSF was 1.08 x 10(8) U/mg, and for huIL-6, it reached 1.95 x 10(7) U/mg. CONCLUSION: huGM-CSF(9-127)-IL-6(29-184) fusion protein with high purity and both huGM-CSF and huIL-6 biologic activities was obtained. PMID- 12901108 TI - [Features of breast ultrasound image and its correlation with estradiol and progesterone level in different phases of menstrual cycle in normal women]. AB - OBJECTIVE: To determine the features of the breast ultrasound image and its correlation with estrogen and progestogen level during different phases of menstrual cycle in normal women. METHODS: Breast ultrasound was performed during luteal phase and late follicular phase in 50 women aged 25-45, with normal menstrual cycle and no distinct breast disease. Breast section thickness, ductal width, breast structure and blood flow were measured. Initial time and persistent days of the cyclic breast pain were recorded. Serum estradiol (E2) and progesterone (P) levels were measured by enzyme immunoassay on the day of ultrasound performed. RESULTS: Breast glandular section was thicker (increasing by 7.3%) in luteal phase than that in late follicular phase and breast ductal was wider (increasing by 13%) in late follicular phase than that in luteal phase. However, both of them had no significant difference. Thirty women (60%) experienced cyclic premenstrual mastalgia, lasting 4 days. Twenty four women (48%) had breast structure changes, nineteen (63.3%) of 30 women with cyclic mastalgia, but only 5 (25%) of 20 women without breast pain had breast structure change (chi 2, P < 0.01). Average serum E2 level of women with or without cyclic breast pain in luteal phase were 365.0 pmol/L and 335.3 pmol/L respectively (P > 0.05), serum progesterone level were 73.3 nmol/L and 66.1 nmol/L respectively (P > 0.05). Serum E2 levels of women with or without cyclic breast pain in late follicular phase were 299.9 pmol/L and 385.8 pmol/L respectively (P = 0.05). Average serum E2 levels of women with or without breast structure change in luteal phase were 368.1 pmol/L and 322.7 pmol/L respectively (P < 0.05). Serum P levels were 78.8 nmol/L and 62.2 nmol/L respectively (P = 0.05); Estradiol levels in late follicular phase were 301.9 pmol/L and 364.2 pmol/L respectively (P > 0.05). CONCLUSIONS: Image of breast ultrasound changed following menstrual cycle in normal menstrual women. Breast structure changes is related to breast pain. Breast pain and breast structure changes may be caused by higher E2 and P level in luteal phase and lower E2/P ratio in late follicular phase. PMID- 12901109 TI - [Effects of lovastatin combined with cyclosporine A on cultured human peripheral blood mononuclear cells proliferation, cytokines expression and cytotoxicity]. AB - OBJECTIVE: To investigate the effects of lovastatin combined with cyclosporine A (CsA) on cultured human peripheral blood mononuclear cells (PBMC) proliferation, cytokines expression and cytotoxicity in vitro. METHODS: The PBMC was isolated by using Ficoll-Hypague density gradient centrifugation method and its viability was evaluated with LDH-release assay. The effects of lovastatin combined with CsA on cultured PBMC proliferation and cytotoxicity were observed in 3H-TdR incorporation assay. The levels of supernatant IFN-gamma and IL-2 were quantitated respectively with RIA and ELISA-SPOT assay. The IL-2 and IFN-gamma mRNA expressions were determined by RT-PCR. RESULTS: Both lovastatin and CsA inhibited PBMC proliferation, expression of IL-2 and IFN-gamma, and NK cell cytotoxicity in a dose-dependent manner. Combination of the two drugs produced synergism. CONCLUSION: Statins combined with cyclosporine A have a synergic effect and can delay the incidence and development of allograft rejection and prolong the graft life-span in animal models and clinical trials. PMID- 12901110 TI - [Relationship between vascular endothelial growth factor expression and angiogenesis in osteosarcoma]. AB - OBJECTIVE: To determine the relationship between angiogenesis and vascular endothelial growth factor (VEGF) expression in osteosarcoma. METHODS: The expression of VEGF was evaluated by immunohistochemical staining of tumor specimens from 33 patients with osteosarcoma. The microvessel density was assessed by immunostaining with CD34 in the most neovascularized area (hot spots). RESULTS: In osteosarcoma, the mean microvessel count of VEGF strong expression tumors was significantly higher than that of VEGF low-expression tumors (Wilcoxon rank sum test, P < 0.05). The close correlation was found between VEGF expression and microvessel density. In contrast, no association was found between Price's grade of osteosarcoma and VEGF expression, as well as microvessel density. CONCLUSION: These observations suggest that VEGF is an important angiogenic factor in osteosarcoma. PMID- 12901111 TI - [Effect of tissue-type plasminogen activator on differentiation of keratinocyte in the human embryonic epiderm]. AB - OBJECTIVE: To explore the regulating roles of tissue-type plasminogen activator (tPA) in the differentiation of keratinocyte in the human epidermis. METHODS: Immunocytochemistry and in situ hybridization were employed to study tPA expression quantitatively and qualitatively in early, middle and late human embryos skin respectively. RESULTS: (1) The expression of tPA was significantly increased in the development of embryonic period, with the highest level in the middle embryonic phase, but the transcription of tPA still kept rather high level in the late human embryonic phase. (2) tPA was mainly localized in the superficial, more differentiated layers of the epidermis. (3) tPA was localized in peripheral cytoplasm of keratinocyte. CONCLUSION: tPA participates the regulation of the keratinocyte differentiation. PMID- 12901113 TI - [Study on the osteoblast and the growth factors of bone tissue engineering]. AB - To build artificial bone with osteoblast and growth factors is one of popular studies on current bone tissue engineering. This paper has reviewed current studies on the function of the growth factors and the resource, isolation and culture of the osteoblast. It also introduces the interaction of the growth factors, and the development in the transgenosis of the relative growth factors. PMID- 12901112 TI - [Relationship between TT virus infection and hepatocellular carcinoma]. AB - OBJECTIVE: To clarify the relationship between TT virus (TTV) infection and the development of hepatocellular carcinoma (HCC). METHODS: TTV was examined in sera of 112 patients with HCC and 95 patients with chronic liver disease (CLD) without HCC. 30 patients without liver disease were examined as control. TTV DNA was detected by the second generation of TTV PCR primers (NG059, NG061 and NG063) after extraction of DNA from 50 microliters serum. The relationship between TTV infection and clinical features of each group patients was analyzed. RESULTS: TTV DNA was detected in 17.86% (20/112) of HCC patients, and this prevalence in HCC patients was not significantly higher than that of TTV DNA in CLD patients (15.78%, 15/95). There were no significant differences in age, gender, tumor factors (size, TNM classification and markers) and other hepatitis virus markers between TTV-positive and TTV-negative in HCC and CLD patients, except history of blood transfusion [(80.00% vs 35.87% (P < 0.05) in HCC patients and 73.33% vs 27.50% (P < 0.05) in CLD patients)] and ALT [(93 +/- 27) IU/L vs (68 +/- 31) IU/L (P < 0.05) in HCC patients]. It seemed that TTV did not do any harm to the liver function of patients who infected with HBV or HCV, except for elevating ALT level in such a kind of HCC patients. CONCLUSION: Although TTV could do harm to the liver function of HCC patients, it does not contribute to the development of HCC from CLD. PMID- 12901114 TI - [Status of screening tests for cervical cancer and its precancerous lesion]. AB - Several screening methods for cervical cancer and its precancerous lesion are reviewed. Cervical cancer screening using visual inspection, colposcopy, oncogenic human papillomavirus DNA testing, liquid-based monolayers and automated Pap smear screening instruments are all potentially valuable when used alone or in combination. Newly developed techniques provide an opportunity to extend practical cervical cancer screening to large population in limited resource areas, and help do more cost-effectiveness of screening tests in high risk population. PMID- 12901115 TI - [Isolation of a kind of endophytic fungus which can produce taxol compounds]. AB - OBJECTIVE: To isolate a kind of endophytic fungus which can produce taxol compounds from the bark of Taxus Cuspidata Sieb et Zucc. METHODS: Endophytic fungus was isolated from the bark of Taxus Cuspidata Sieb et Zucc by aseptic techniques and the analysis of the culture fluid from the fermentation of fungus was carried out by high performance liquid chromatography (HPLC). RESULTS: Endophytic fungus was obtained from the bark of Taxus Cuspidata Sieb et Zucc and the chemical analysis showed that the culture of fungus contain taxol compounds. CONCLUSION: Endophytic fungus which can produce taxol compounds exists in the bark of Taxus Cuspidata Sieb et Zucc. PMID- 12901116 TI - [Design and application of Chinese universal spinal instrumentation in spine surgery]. AB - OBJECTIVE: To report the rationale and primary results of application of Chinese universal spinal instrumentation (CHN-USI) in spinal surgery. METHODS: From September 1998 to June 2000, 70 patients with scoliosis, including 30 congenital scoliosis, 36 idiopathic scoliosis and 4 other disorders, were treated with CHN USI in our hospital. All patients got follow-up after surgery with roentgenogram. RESULTS: The average follow-up time was 8.9 months. On average, the primary scoliotic curve was reduced from 70.5 degrees (40 degrees-103 degrees) to 32.5 degrees with an average correction rate of 55.6%. The average increases in body height and the length of corrective segment were 6.5 cm and 5 mm respectively. The kyphotic curve was corrected from average 80.5 degrees to 29 degrees. No serious complication was found in our group. CONCLUSIONS: The CHN-USI which has no need of spinal fusion and will elongate together with the children's growth is a new effective instrumentation for correcting scoliosis, especially in case of growing children. It can also be used in the treatment for kyphosis, anterior derotation spondylodesis, spondylolisthesis and spinal fracture. PMID- 12901117 TI - [Application of diapason instrumentation for treatment of unstable lumbosacral spinal stenosis]. AB - OBJECTIVE: To observe the preliminary clinical outcome of using a new instrumentation Diapason system and to introduce the characterization and surgical technique of this new system for treatment of unstable lumbar spinal stenosis. METHODS: 16 patients with unstable spinal stenosis who were treated by decompression, posterolateral intertransverse arthrodesis and transpedicle instrumentation of Diapason system, were analyzed retrospectively. RESULTS: Lower back pain (LBP) of 16 patients were significantly alleviated after surgery (scores of LBP before operation: 47.5 +/- 0.8; scores of LBP after operation: 31.9 +/- 2.3, P < 0.001). There was no implant failure, no early or later infection and no neurological complications in 16 patients at an average of 6.2 month follow-ups. No pseudoarthrosis was observed on roentgenography. CONCLUSION: Our short-term follow-up and limited cases study showed satisfactory preliminary result of treating unstable lumbar spinal stenosis with Diapason internal fixation. PMID- 12901118 TI - [Significance and application of nuclear matrix protein 22 in detecting bladder cancer]. AB - OBJECTIVE: To investigate the significance of nuclear matrix protein 22 (NMP22) in the detection of bladder cancer. METHODS: The level of NMP22 was measured in the urine of 90 patients (40 patients with bladder cancer and 50 patients with non-bladder cancer) by ELISA and compared with urinic cytology (UC). RESULTS: The level of NMP22 in bladder cancer group was significantly higher than that of control group, and it was not influenced by hematuria. The sensitivity of NMP22 was high, but the specificity was lower than that of UC. CONCLUSION: NMP22 can be used to screen and follow up bladder cancer with high sensitivity. PMID- 12901119 TI - [Diagnosis and treatment of primary intracranial lymphoma]. AB - OBJECTIVE: To discuss the clinical characteristics, diagnosis, and treatment of primary intracranial lymphoma. METHODS: A total of 148 cases of primary intracranial lymphoma from January 1984 to January 2000 were reviewed in our study. RESULTS: Among the 148 cases, total and subtotal removal was achieved in 117 cases, partial removal in 31 cases. All the cases were treated by the radiotherapy after surgery. 44 cases combined with chemotherapy. The symptoms of patients in 112 cases (76%) were relieved and 36 cases (24%) progressed with 8 cases dead after surgery. CONCLUSION: Primary intracranial lymphoma is a malignant neoplasm. The main treatment is surgical removal combining with postoperative radiotherapy. Prognosis is poor. PMID- 12901121 TI - Recombinant factor VIIa: a universal hemostatic agent? AB - Recombinant factor VIIa (rVIIa) has proved effective for the treatment and prevention of hemorrhage in patients with inherited hemophilia A and B who develop inhibitors to factor VIII or IX, and patients with acquired hemophilia A. More recently, there is evidence that rVIIa may also be effective in the control of abnormal bleeding in a variety of other conditions, such as inherited factor VII deficiency, thrombocytopenia, Glanzmann's thrombasthenia, and liver disease. In some of the reports, rVIIa appeared to be effective in controlling massive hemorrhage in which there was no response to conventional measures. It is now considered by some to be potentially the first universal hemostatic agent. However, further prospective, controlled, and adequately powered clinical studies are clearly required. It will be of particular interest to determine the efficacy of rVIIa in conditions such as severe thrombocytopenia, severe von Willebrand disease, severe defects in platelet activation, and severe deficiencies of factors V, X, II, and fibrinogen in which effectiveness would seem to be unlikely based on our current understanding of mechanisms of action of rVIIa. PMID- 12901120 TI - Evaluation of abnormal bleeding in women. AB - The evaluation of the woman with abnormal bleeding, particularly menorrhagia, carries a relatively high yield of discovering an underlying disorder of hemostasis. The most common underlying hemostatic disorder would be von Willebrand disease, with an estimated prevalence of 7% to 20%. In addition, preliminary results suggest that another 20% to 30% of patients may have impaired platelet aggregation as another cause of menorrhagia. Disorders of fibrinolysis may be an additional underlying hemostatic disorder. Initial intake should include documentation of menorrhagia by the pictorial chart assessment of menstrual flow. Baseline characteristics of menstrual flow should also be documented including the frequency of changing the sanitary napkin on the heaviest day, use of more than one sanitary napkin at a time, and number of days lost from school or work. Menorrhagia since menarche, a history of surgicalor dental-related bleeding, and a history of postpartum hemorrhage are items of the bleeding symptom audit that appear to predict in part von Willebrand disease in women with menorrhagia. Epistaxis and easy bruising do not appear to be clearly discriminatory symptoms. Initial testing should include the complete blood cell count, protime, activated partial thromboplastin time, iron profile, serum creatinine, thyroid-stimulating hormone level, factor VIII level, von Willebrand factor antigen, ristocetin cofactor, and platelet aggregation studies. Additional hemostatic studies may also include a factor XI level and euglobulin clot lysis time. Intuitively, failure to diagnose an underlying hemostatic disorder may lead to continued menorrhagia and diminished quality of life, as well as unnecessary surgical interventions, which, in turn, may be fraught with increased bleeding. PMID- 12901122 TI - Acquired hemophilia: diagnosis and management. AB - In this review, we describe our current understanding of the mechanisms of autoimmunity, the effect of antibodies on factor VIII, and immunosuppressant agents currently in use. Factor VIII autoantibodies may arise because of dysregulation of the immune system leading to increased autoreactivity or to clonal expansion of reactive B cells. Patients generally have several antibodies directed against a variety of functional epitopes. These antibodies may inhibit binding of factor VIII to the von Willebrand factor or phospholipid, prevent the association of factor VIII with factor IXa, block the binding of factor Xa to factor VIII, or interfere with the formation of the factor VIIIa-factor IXa phospholipid (tenase) complex. Additionally, antibodies have been described that catalyze the hydrolysis of factor VIII. The production of antibodies has been effectively suppressed by a number of drugs, including steroids, cyclophophamide, cyclosporine, and rituximab. Beginning with a short course of steroids and then adding one or more agents as needed is a currently accepted strategy for restoring normal factor VIII levels. PMID- 12901123 TI - The diagnosis and management of disseminated intravascular coagulation. AB - This review describes disseminated intravascular coagulation (DIC) as a syndrome in which hemostatic factors are activated. The syndrome ranges in severity from a decompensated coagulopathy (overt-DIC) to the subclinical compensated activation of hemostatic factors (nonovert DIC). The first part of this review emphasizes two points. First, activation of the hemostatic system is controlled by a vast network of capillaries and venules through anticoagulant and anti-inflammatory regulatory factors that operate from the endothelium (e.g., protein C and thrombomodulin, tissue factor pathway inhibitor). These hemostatic regulators can be overridden by procoagulant disorders such as amniotic fluid embolism or degraded by proinflammatory disorders such as sepsis. Second, because this link between the microvascular endothelium and circulating hemostatic factors is so close, even a relatively mild disturbance of the microvasculature targeted by the inflammatory process may be reflected systemically by changes in molecular biomarkers of hemostatic activity. Therefore, application of criteria for the diagnosis of nonovert DIC should be of value in detecting a compensated response to inflammatory stress of the microvasculature in patients who are at risk before they develop an uncompensated over DIC response and organ failure. The second part of this review covers the recent experience investigators have had in diagnosing and following the response of patients to treatment with biomarkers. PMID- 12901124 TI - Risk factors for bleeding in patients taking coumarins. AB - Coumarins (e.g., warfarin) are used for the treatment and prophylaxis of venous and arterial thromboembolism. Unfortunately, their use is limited by bleeding. In deciding whether to initiate coumarin therapy, the therapeutic benefits must be weighed against the potential risk of bleeding for each individual patient. The major determinants of coumarin-related bleeding are the intensity of anticoagulant effect as measured by the International Normalized Ratio, patient characteristics, concomitant use of other drugs, and the length of anticoagulant therapy. At the initiation of coumarin therapy, the risk factors for bleeding can be identified by the COUMARINS acronym and by one of the bleeding prediction models. During ongoing therapy with coumarins, the risks and benefits of therapy should be reassessed periodically. This article reviews the current evidence for the prediction and prevention of coumarin-related bleeding. PMID- 12901125 TI - Hypercoagulable states and pregnancy. AB - Pulmonary thromboembolism is a major cause of maternal death in the developed world. Underlying this condition is the problem of deep venous thrombosis. These problems often reflect an underlying hypercoagulable state. Growing evidence implicates congenital and acquired thrombophilias in the pathophysiologic processes underlying not only venous thrombosis, but also miscarriage, intrauterine growth restriction, and preeclampsia, raising new possibilities for intervention. PMID- 12901126 TI - Heparin-induced thrombocytopenia. AB - Heparin-induced thrombocytopenia (HIT) is a syndrome of antibody-mediated activation of platelets and coagulation that confers increased risk for thrombosis. This article reviews recent studies of the pathogenesis, laboratory testing, frequency, and clinical presentation of HIT. Topics discussed include the nature of the neoepitopes on platelet factor 4 recognized by HIT antibodies, the nonspecific nature of the polyanions that support neoepitope formation, activation of monocytes and endothelium, and the development of new animal models. The transient "autoimmune" nature of HIT is highlighted in relation to new data concerning the rapid decline in HIT antibodies, as well as the recent recognition of a syndrome of "delayed-onset HIT" in which thrombocytopenia and thrombosis begin several days after stopping heparin. Detecting only HIT antibodies of the immunoglobulin G class is suggested as one way to increase diagnostic specificity of laboratory testing. Recent reports suggest that HIT could explain approximately 5% of cases of acute adrenal failure caused by bilateral adrenal hemorrhagic infarction. PMID- 12901127 TI - Deep venous thrombosis in the medically ill. AB - Epidemiologic data have shown that the risk of venous thromboembolism (VTE) in medical patients is comparable to the risk in surgical patients. Moreover, autopsy studies demonstrated that 75% of all in-hospital deaths for pulmonary embolism occur in nonsurgical patients. Hospitalized medical patients represent a very heterogeneous population. Patients with medical disorders who are at significant risk include those with heart failure, acute respiratory failure, acute infectious diseases, acute rheumatic disorders, inflammatory bowel diseases, and cancer. Recent consensus conferences have produced detailed recommendations for thrombosis prophylaxis in both surgical and medical patients. Both unfractionated heparin and low-molecular-weight heparins (LMWHs) have been shown to be effective when compared with placebo and are recommended in high-risk medical patients. When LMWHs were compared with unfractionated heparin, they were shown to have equivalent efficacy and greater safety than the parent compound. As for surgical patients, routine prophylaxis is still underused in the general medical population. PMID- 12901128 TI - Management of thrombosis in patients with hematologic malignancies. AB - The management of thrombosis in patients with cancer is a clinical challenge. Although the prevalence of thromboembolic complications in hematologic malignancies is lower than that reported for solid tumors, thrombotic events are increasingly being reported both in patients with acute leukemia and in those with chronic myeloproliferative and lymphoproliferative disorders. Accordingly, there is the need for promptly identifying patients at a higher risk of developing thromboembolic complications, as well as instituting appropriate measures of prophylaxis and treatment. We briefly summarize the pathogenetic evidence for the existence of a prothrombotic state in patients with hematologic malignancies, discuss common clinical pictures, and examine those modalities of prophylaxis and treatment that are commonly recommended in this clinical setting. Further work is needed to provide physicians with specific guidelines for a systematic management of thrombosis in patients with hematologic malignancies. PMID- 12901129 TI - Risk of deep venous thrombosis in medical patients. PMID- 12901130 TI - Neutrophil growth factors. AB - A significant advance in the field of neutrophil growth factors has occurred with the commercial availability of pegfilgrastim (Neulasta, Amgen, Thousand Oaks, CA), a new-generation, pegylated filgrastim molecule with a sustained duration of action. Pegylation of filgrastim allows once-per-chemotherapy cycle frequency of administration, in contrast to repeated daily administration of filgrastim. Clinical data from two randomized trials demonstrate equivalence of pegfilgrastim and filgrastim in duration of severe neutropenia and recovery from absolute neutrophil count nadir following myelosuppressive chemotherapy. In addition, secondary endpoint results in both trials suggest an enhanced reduction in the overall incidence of febrile neutropenia with pegfilgrastim. Neutrophil kinetic studies demonstrate steady serum neutrophil levels following pegfilgrastim administration, in contrast to the peak-and-trough neutrophil effects observed following filgrastim administration. Granulocyte colony-stimulating factor (G CSF) therapy has an antiapoptotic effect on neutrophils, which may be enhanced by continuous serum concentrations of pegfilgrastim. Monocytes possess a G-CSF receptor, and this finding has fueled investigational analysis of the role of G CSF as a mediator in the host inflammatory response to foreign pathogens. The data demonstrate that depending on the timing of administration, G-CSF may function as a proinflammatory mediator or an anti-inflammatory mediator. It is likely that the early, prophylactic administration of pegfilgrastim creates an environment in which an anti-inflammatory response predominates. Additional investigational studies will be necessary to confirm and better define the mechanism for enhanced benefit of pegfilgrastim over filgrastim. The recent biologic findings of the mechanism of G-CSF therapy reviewed here provide a strong basis from which further research initiatives may be conducted. PMID- 12901131 TI - Stem cell transplantation and hematopoietic growth factors. AB - Hematopoietic growth factors are commonly used in allogeneic and autologous stem cell transplantation. The growth factors most frequently used are human recombinant erythropoietin, filgrastim, and sargramostim, and a number of trials have been done using them either singly or in various combinations for mobilization, post-transplant, and for delayed engraftment. Filgrastim and sargramostim can shorten the neutropenic period and decrease infectious complications post-transplant, thus lowering the cost of both autologous and allogeneic transplants. Erythropoietin has not been particularly effective for mobilization, and studies have not shown its efficacy in reducing red blood cell transfusions in autologous transplants. However, they have been clinically beneficial in allogeneic transplantation and in delayed erythropoiesis post transplantation. Stem cell factor remains investigational at this time but seems promising. The new long-acting erythropoietin and filgrastim are also introduced here and briefly discussed. PMID- 12901132 TI - Challenges in the development of platelet growth factors: low expectations for low counts. AB - Thrombocytopenia remains a significant clinical problem for which only symptomatic therapy, namely platelet transfusion, is available for management of acute events. Platelet transfusions are often complicated by febrile reactions, as well as the risk of transmission of infectious agents and the likelihood of alloimmunization, which then decreases the effectiveness of additional transfusion support. The availability of a hematopoietic cytokine that could reliably stimulate platelet recovery, analogous to the effect of granulocyte colony-stimulating factor on neutrophil recovery following chemotherapy, would greatly enhance supportive care in cancer and provide an effective therapy for a variety of diseases that cause thrombocytopenia. To identify such a thrombopoietic cytokine, studies initially focused on regulatory molecules that stimulates early multipotent hematopoietic progenitors, such as interleukin-1 and interleukin-6. Unfortunately, these cytokines had poor efficacy and significant toxicity in human testing. Recombinant human interleukin-11, an early-acting cytokine, has modest efficacy and clinically challenging toxicities, but in the absence of other active drugs, it has been licensed for prevention of severe chemotherapy-induced thrombocytopenia. Recent interest has focused on analogs of thrombopoietin, the endogenous regulator of thrombopoiesis, which have the potential for much greater efficacy with minimal toxicity due to the more specific targeting of megakaryocyte-specific signaling. PMID- 12901133 TI - Red cell growth factors in patients with chronic anemias. AB - Erythropoetin therapy has been approved for treatment of medical anemia since 1989. The adoption of this strategy has been rapid in some settings (e.g., renal failure patients) and progressive in others (e.g., cancer patients). Although the risks of blood transfusion have declined substantially, risks associated with (untreated) anemia have undergone new scrutiny. Options such as novel erythropoiesis-stimulating protein bring new alternatives to blood transfusion. Erythropoietin therapy is undergoing renewed scrutiny in the management of anemia, not only because of traditional concerns regarding blood risks but because emerging evidence suggests that improved patient outcomes result from management of anemia. PMID- 12901134 TI - Innovations in blood donor screening and blood collection. AB - Blood donor screening and blood collection are two of the donor-focused activities inherent in the process of maintaining an adequate blood supply. Donor screening represents procedures to provide safety for both donors as well as recipients. Donor medical history questions are so cumbersome that self administered manual, computer-assisted, or on-line donor questions are under consideration in order to reduce time involved in the screening process. Blood collection procedures remain primarily based on whole blood donations, but apheresis donations are increasing steadily. The yield of multiple transfusible units per donation, as well as quality and reproducibility per product, support the use of apheresis procedures or other automated whole blood techniques to help offset blood shortages. PMID- 12901135 TI - Molecular biology in transfusion medicine: current applications and future practice. AB - Alloantibodies to blood group antigens are produced because there are differences between the blood group antigens on recipient and donor (or mother and fetus) red blood cells. Historically and currently, blood group antibodies and antigens are detected by hemagglutination, which can require considerable skill. Over the past 2 decades, there has been an astounding pace of growth in the field of molecular biology techniques and even more recently in the understanding of the basis of many blood group antigens and phenotypes. Thus, we are now able to consider identification of blood group antigens in genetic terms and identification of blood group antibodies using molecular approaches. Indeed, this knowledge is being applied to help resolve some long-standing clinical problems that cannot be resolved by classic hemagglutination. This article reviews our current knowledge of molecular approaches for identifying blood group antigens and antibodies as applied to transfusion medicine. PMID- 12901136 TI - The red blood cell transfusion threshold: evidence and outcome. AB - The appropriate threshold for administration of red blood cell transfusion is the subject of controversy. Recently, several prospective randomized clinical trials have compared restrictive and liberal transfusion regimens in the settings of critical care and surgery. In addition, several large cohort studies have provided information on associations between anemia, transfusion, and clinical outcomes. Taken together, these studies generally support conservative red blood cell transfusion strategies. PMID- 12901137 TI - Noninfectious serious hazards of transfusion. AB - Serious, noninfectious transfusion complications include transfusion-related acute lung injury (TRALI), transfusion-associated graft-versus-host disease, anaphylaxis, hemolysis, and post-transfusion purpura. Prompt recognition and treatment are crucial, but prevention is preferable. Many transfusion reactions are not recognized as such, perhaps because signs and symptoms mimic other clinical conditions. However, any unexpected symptoms in a transfusion recipient should at least be considered as a possible transfusion reaction and be evaluated. Appropriate diagnosis is the key to treatment and may prevent additional reactions, not only in the patient, but possibly, as in the case of TRALI, in other patients. PMID- 12901138 TI - Infectious risks of blood transfusion. AB - The public demand for a "zero-risk" blood supply in the wake of transfusion transmitted HIV has prompted many changes in the donor selection process, donor testing, and postcollection processing of blood and blood products. The effectiveness of these new strategies has had a remarkable impact: The risk for HIV is now estimated to be one in 1 million units transfused. The risk for other transfusion-transmitted viral infections has similarly been reduced. On the heels of the success of research endeavors targeting transfusion-related viral risk reduction, new concerns regarding transmission of other infectious agents (e.g., bacteria, parasites, and prions) by transfusion have emerged. Consequently, investigative efforts that are even more vigorous are under way, aiming to define the characteristics of transmission as well as identify useful technologies to detect or eliminate these infectious agents. PMID- 12901140 TI - Erythrocyte adhesion in sickle cell disease. AB - The vaso-occlusive process in patients with sickle cell disease is complex and is likely to involve interactions between hemoglobin S red blood cells (SS RBCs) and vascular endothelium, as well as between SS RBCs and leukocytes. Vaso-occlusive events lead to recurrent pain and a wide spectrum of end-organ damage, including pulmonary hypertension and renal failure. However, the triggers inducing adhesion and vaso-occlusion are only now being elucidated. Investigators have characterized the ability of a number of RBC surface structures to adhere to both endothelial cells and components of the subendothelial extracellular matrix. In addition, evidence is accumulating to suggest that SS RBC adhesion receptors undergo activation under physiologic conditions. An understanding of these mechanisms at the molecular level should ultimately allow development of new preventive and treatment strategies to abrogate vaso-occlusive events. PMID- 12901139 TI - Therapies to increase fetal hemoglobin in sickle cell disease. AB - The pathophysiology of sickle cell disease originates in the polymerization of sickle hemoglobin. Fetal hemoglobin (HbF) inhibits this process, and high HbF concentrations reduce the severity of the disease. Drugs with presumed mechanisms of action that perturb the orderly maturation of erythroid precursor cells, induce hypomethylation of HbF genes, bind transcriptionally active elements of these genes, or influence chromatin structure can enhance HbF production in sickle cell disease. Hydroxyurea, a drug that affects erythroid maturation and regeneration, reduces morbidity and mortality in adults with sickle cell anemia. Its use in young children and in combination with other classes of HbF-inducing agents is being studied. PMID- 12901141 TI - Update on the clinical use and misuse of erythropoietin. AB - Anemia is a common finding in patients with hematologic malignancies and most commonly can be attributed to the anemia of chronic disease compounded by the myelotoxic effects of chemotherapy. Symptoms of anemia include fatigue, and the patient's quality of life may be impaired. Possible treatments for the anemia are to do nothing, to transfuse with red cells, or to treat with recombinant human erythropoietin (rhEPO). rhEPO has become standard treatment for the anemia in chronic renal failure and has been successfully used in anemia secondary to malignancy. In patients with lymphoproliferative diseases, rhEPO increases the hemoglobin concentration, decreases the need for transfusion, and improves the patients' quality of life. Disadvantages of rhEPO include its cost, efficacy in only around 60% of patients, and delay of 4 to 8 weeks before maximum benefit is achieved. The anemia in patients with myelodysplasia responds less well to rhEPO. Misuse of rhEPO is common in the clinical setting but usually not of clinical importance. Misuse to enhance sporting prowess is probably rare but has potentially serious consequences. PMID- 12901142 TI - Recent developments in the anemia of chronic disease. AB - The anemia of chronic disease (ACD) is a hypoproliferative anemia defined by a low serum or plasma iron concentration in the presence of adequate reticuloendothelial iron stores. It is been established that ACD results from the effects of cytokines that mediate the immune or inflammatory response. During the past 3 years, the clinical scope of this syndrome has been expanded beyond the traditional chronic infectious, inflammatory, or neoplastic diseases to include other, often acute, syndromes in which the same pathogenetic mechanisms are operating. An improved understanding of the use of the soluble transferrin receptor concentration in clinical medicine has enhanced the ability to diagnose ACD, and further experience with the use of recombinant human erythropoietin in the management of severely affected patients with ACD has provided a basis for rational and effective management. Ongoing studies of the mechanisms contributing to the development of ACD continue to elucidate the pathogenesis of this common and clinically significant syndrome. PMID- 12901143 TI - Approach to anemia associated with myelodysplastic syndromes. AB - Severe and transfusion-dependent anemia is a major symptom for patients with myelodysplastic syndromes (MDSs). Recent translational research studies have improved treatment approaches for these patients and contributed to our understanding of the pathophysiology of MDSs. For example, the response to antithymoglobulin treatment in certain patients with refractory anemia has elucidated potential immune mechanisms underlying bone marrow failure, whereas the observed effects of erythropoietin with or without granulocyte colony stimulating factor in other low-risk patients have offered insights into the role of mitochondria-mediated erythroid apoptosis in MDS-related anemia. Current research is aimed at defining the role for other potential pathogenetic mechanisms in the development of MDS. Such studies may lead to individually tailored therapy for MDS patients based on a detailed analysis of the role of different contributing mechanisms to the origins of their disease. PMID- 12901144 TI - Pathogenesis of thrombotic thrombocytopenic purpura. AB - The recent discovery of important molecular and genetic mechanisms of thrombotic thrombocytopenic purpura and hemolytic uremic syndrome provide an opportunity to reconstruct scientific and clinical paradigms. Acquired and congenital defects of the metalloprotease ADAMTS13 are a central feature in the pathogenesis of a major type of thrombotic microangiopathy. This and other pathogenic mechanisms can redefine the terminology of thrombotic microangiopathy. Deficient activity of ADAMTS13 suggests several possible models of microvascular thrombosis. The sporadic relationship between thrombotic microangiopathy and ADAMTS13 deficiency draws attention to other critical pathologic factors that are still poorly understood. Investigation of vascular injury and mechanisms of microvascular thrombosis remain the frontiers of investigation in thrombotic microangiopathy. PMID- 12901145 TI - Management of thrombocytopenic bleeding: is there a role for recombinant coagulation factor VIIa? AB - The ability of high concentrations of recombinant factor VIIa (rFVIIa) to improve thrombin generation in in vitro and ex vivo models of thrombocytopenia, and to improve bleeding time in thrombocytopenic animal models supports a potential role for rFVIIa in the management of thrombocytopenic bleeding. Anecdotal reports suggest that rFVIIa is effective in the prevention and treatment of bleeding episodes in surgical procedures in a limited number of patients with severe thrombocytopenia. There is a need for clinical trials to determine whether rFVIIa is efficacious and safe in severe thrombocytopenic bleeding, either by itself or in combination with other hemostatic agents such as platelet transfusion. Pending the availability of such data, rFVIIa may be considered in severe thrombocytopenic bleeding that is unresponsive to human leukocyte antigen compatible platelet transfusions and other standard treatments. If rFVIIa is used, unresolved issues include optional dosage, dosing interval, and whether rFVIIa is best used alone or in conjunction with platelet transfusion. PMID- 12901146 TI - Laboratory diagnosis of immune heparin-induced thrombocytopenia. AB - Immune heparin-induced thrombocytopenia (HIT) is a distinct immunohematologic syndrome in which laboratory detection of the pathogenic HIT antibodies is diagnostically useful. Assays can be broadly classified as platelet activation assays (which detect HIT antibodies based on their characteristic platelet activating properties) and antigen assays (which measure antibodies reactive against platelet factor 4 complexed with heparin or other polyanions). Available tests vary considerably in their sensitivity-specificity profiles for detecting the antibodies and in their predictive values. The high sensitivity of certain assays means that HIT can be readily ruled out (high negative predictive value). However, because heparin-treated patients often generate nonpathogenic antibodies, a positive test does not necessarily indicate clinical HIT. Laboratory methods for detecting HIT antibodies have undergone an evolution similar to that of red blood cell serology and which parallels the Goldilocks tale, that is, progression from too insensitive (too hard) to too sensitive (too soft) to "just right." However, optimal diagnostic information requires that laboratory test results be interpreted in the appropriate clinical context. PMID- 12901147 TI - Strategies for managing refractoriness to platelet transfusions. AB - The availability of platelet transfusions made possible intensive, myelosuppressive therapy. Chronic transfusion support is often complicated by refractoriness to platelet transfusions characterized by inadequate post transfusion platelet count increments. Refractoriness is most commonly associated with a variety of clinical factors. Aggressive treatment of these clinical conditions often corrects the response to platelet transfusions. Fewer patients develop immune-mediated platelet destruction associated with alloantibodies to class I human leukocyte antigens (HLAs) and occasionally to platelet-specific antigens. The majority of these patients are successfully supported with HLA matched or cross-match compatible single-donor platelet concentrates. PMID- 12901149 TI - Aplastic anemia--low drug associations. PMID- 12901148 TI - Platelet substitutes in the management of thrombocytopenia. AB - A variety of agents can improve hemostasis and reduce blood loss caused or exacerbated by congenital or acquired defects in platelet number of function. This review discusses several available approaches for the practicing clinician to reduce bleeding (diffuse or localized) in this clinical setting. PMID- 12901150 TI - Rituximab for follicular lymphoma. AB - Treatment with the chimeric anti-CD20 antibody rituximab has been rapidly accepted into the clinical treatment of patients with CD20 positive lymphoma. The low toxicity profile, relative ease of administration, and encouraging response rates observed as a single agent allow it to be used alone or in combination with or following standard chemotherapies. Patients with follicular non-Hodgkin's lymphoma (NHL) have a high response rate to treatment with this new modality, with overall response rates of 50% to 60% in the relapsed setting and 70% in the initial setting. The addition of scheduled retreatment or maintenance therapy leads to improved clinical responses and delays time to progression. Combination trials with chemotherapy demonstrate feasibility and promising response rates including clearing of minimal residual disease detected using molecular techniques. To date, there are limited randomized clinical trial data available to guide the use of this new modality in this patient population and few long term results. It remains difficult to determine when and how to use this new modality best in the overall treatment course of these patients. This paper discusses the rationale for the use of rituximab in patients with follicular NHL and discusses the available data involving dosing, schedule, timing, and combinations with chemotherapy. PMID- 12901151 TI - Monoclonal antibodies in the management of newly diagnosed, aggressive B-cell lymphoma. AB - Monoclonal antibodies have been used as a therapeutic option for patients with lymphoma for less than 5 years, but they have already changed our perception of how to treat these patients. In this review, the use of different types of monoclonal antibodies in the treatment of patients with diffuse large B-cell lymphoma is analyzed, particularly the combination of rituximab with chemotherapy as a first-line therapeutic option. The putative role of other monoclonal antibodies is discussed. PMID- 12901152 TI - Radioimmunotherapy of relapsed or refractory low-grade, follicular, or transformed B-cell non-Hodgkin's lymphoma. AB - Radioimmunotherapy consists of radiolabeled monoclonal antibodies for the treatment of malignancy. For more than a decade, radioimmunotherapy has shown great promise for the treatment of B-cell lymphoma. During the past decade, two products targeted to the CD20 antigen on B cells, iodine-131 tositumomab and yttrium-90 (90Y) ibritumomab tiuxetan, have been tested extensively in registration trials for potential licensing approval by the US Food and Drug Administration (FDA). Both products have produced response rates of 70% to 80% in low-grade and follicular lymphoma, and response rates of 50% to 60% in low-grade or follicular lymphoma that has transformed into an intermediate or high-grade lymphoma. Median duration of response to a single course of treatment has been about 1 year, with complete remission rates in one quarter to one third of patients. In February, 2002, 90Y ibritumomab tiuxetan was formally approved by the FDA for the treatment of relapsed or refractory low-grade, follicular, or transformed B-cell lymphoma, including rituximab-refractory transformed lymphoma, thus becoming the first radioimmunotherapeutic agent approved by the agency. The product became available for commercial use in April, 2002. Clinical trials will determine how radioimmunotherapy will be integrated into the treatment of lymphoma. PMID- 12901153 TI - Clinical applications of molecular monitoring in leukemia. AB - Molecular analyses of the leukemias have been able to identify unique markers that can be used as "molecular signatures" for the various diseases. In acute promyelocytic leukemia, qualitative polymerase chain reaction (PCR) has helped define the clinical syndrome and has been used to guide therapy. Quantitative modifications of the PCR technique have been investigated in chronic myelogenous leukemia and acute lymphocytic leukemia and found to correlate with clinical outcomes. These assays may soon be incorporated into standard clinical management and may result in a new definition of response. PMID- 12901154 TI - Nonmyeloablative allogeneic stem cell transplantation for acute leukemia. AB - Allogeneic stem cell transplantation (ASCT) has traditionally included the administration of maximally tolerated doses of chemoradiotherapy, which have been associated with significant treatment-related toxicities. Thus, less intensive conditioning regimens have been explored as a safer alternative to conventional ASCT. Nonmyeloablative stem cell transplantation (NMSCT) has been one of the most promising recent developments in the treatment of hematologic malignancies, and early studies have yielded encouraging results with high engraftment rates and sustained remissions. This approach incorporates immunosuppressive doses of chemotherapy and radiotherapy to achieve a mixed donor-host hematopoietic chimeric state and allows the development of a donor immune-mediated graft-versus leukemia effect as the primary means of disease eradication. This review discusses the background and rationale behind NMSCT and its impact on the treatment of patients with acute leukemia. PMID- 12901156 TI - Management of advanced chronic lymphocytic leukemia. AB - Chronic lymphocytic leukemia (CLL) is generally considered to be incurable. It is the most common of the adult leukemias, and many patients are asymptomatic when diagnosed. Most patients survive for several years, and some never require treatment for their disease, leading to the belief that it is an indolent disease. However, this view is changing with the introduction of more effective therapies. Purine nucleoside analogs have become standard in the therapy of most patients. Monoclonal antibodies including alemtuzumab (Campath-IH; Berlex Laboratories, Richmond, CA) and rituximab are playing an increasingly important role in the treatment of patients with advanced disease. Clinical trials will be crucial in defining how and when to treat patients with CLL and will help establish the role of newer prognostic markers and more sensitive methodologies to detect minimal residual disease. PMID- 12901155 TI - Practical aspects of the treatment of chronic myelogenous leukemia with imatinib mesylate. AB - Imatinib mesylate (Gleeve; formerly known as STI-571; Novartis Pharmaceuticals, Basel, Switzerland) is a protein tyrosine kinase inhibitor that has been approved by the US Food and Drug Administration for chronic-phase chronic myeloid leukemia (CML) that is refractory to or intolerant of interferon-alpha, and with accelerated and blast-phase CML. Imatinib is remarkably effective as treatment for chronic-phase and accelerated-phase CML at doses of 400 mg and 600 mg daily, respectively. Although dramatic responses are noted in patients with myeloid blast crisis, lymphoid blast crisis, and Philadelphia chromosome-positive acute lymphoblastic leukemia, the responses are usually incomplete and of short duration. The role of imatinib in relation to allogeneic stem cell transplantation is discussed, recognizing that the data on which any decisions can be made are relatively immature. At this time, it remains to be seen whether the chronic phase of CML can be extended sufficiently long in some patients so that they are functionally "cured," and whether doses of imatinib higher than 400 mg daily will improve response rates in chronic-phase CML. The feasibility of combining imatinib with other agents also needs to be examined, as does the utility of molecular monitoring of response. Finally, developing methods to overcome resistance to imatinib is a looming challenge. PMID- 12901157 TI - Monoclonal antibody therapy in acute myeloid leukemia. AB - Monoclonal antibody therapy for patients with acute myeloid leukemia is currently based on targeting cell surface receptors selectively expressed on myeloid cells. These include primarily CD33 and CD45. Antibodies are unlabeled or have been conjugated to various radioisotopes or DNA-damaging cytotoxic agents. The safety of this approach has been determined, and antibody-related infusional events are common. Other toxicity is primarily dependent on the conjugate to which the antibody has been linked. Efficacy of this approach is still to be determined. Phase II studies have demonstrated antileukemic responses with all agents, although less so with unlabeled antibodies. Whether the use of these antibodies in combination with, or as a substitute for, currently available therapy will lead to improved outcomes for patients with acute myeloid leukemia has not been demonstrated to date. PMID- 12901158 TI - Diagnosis and management of core-binding factor leukemias. AB - The core-binding factor (CBF) leukemias are the subtype of acute myelogenous leukemia (AML) associated with the highest response to therapy and longest remission duration. Although CBF leukemias have two distinctly different morphologic and clinical presentations (i.e., AML M2 and AML M4eo), their molecular pathophysiology is linked by their abnormalities in different subunits of CBF, a transcription factor important in several pathways in hematopoietic cells. This article summarizes the recent biologic and clinical advances made in the treatment of these diseases. PMID- 12901159 TI - The ultimate shortage. PMID- 12901160 TI - Aetna agrees. PMID- 12901161 TI - A booster for vaccines. PMID- 12901162 TI - Filling a need. PMID- 12901163 TI - Stopping an unwelcome visitor. PMID- 12901164 TI - Female suicides in major Texas cities, 1994 through 1998. AB - From 1976 to 1985, Texas ranked first among states in the proportion of injury deaths caused by firearms, with an annual rate of 21.2 per 100,000. From 1995 to 1997, suicide was the eighth leading cause of death in Texas, claiming 2137 lives and accounting for 20% of all injury deaths in Texas. Firearm injuries significantly affect mortality rates in many states. The rates of suicide have continued to rise, particularly in young adults, and the rate of suicide by firearms has also increased significantly throughout the United States. We have focused on the most common method of suicide by age and race among women in Texas from 1994 through 1998. This work extended that of Li et al on suicide in Austin, Travis County, Texas, from 1994 through 1998. Those investigators found that the most common method of suicide in females was by firearms. In this current study, we asked whether the Austin data for females were unique or were representative of other major cities in Texas, including Dallas, El Paso, Houston, and San Antonio. Our major finding was that the use of firearms was the most common method of suicide among women in all of these cities. Women aged from 35 to 44 years had the highest incidence of deaths, followed by those aged from 45 to 54 years. Hispanic women had the lowest rate of suicide compared with African American and white women. Overall, rates of suicide among white males and females are at or higher than the US average in all Texas cities examined, except Dallas. Suicide rates among African-American males are at or below the US average, whereas the suicide rate for African-American females exceeds the national average. Nationwide rates for Hispanics were not available. PMID- 12901165 TI - Chromium in intertidal sediments of the Clyde, UK: potential for remobilisation and bioaccumulation. AB - Detailed surveys of intertidal sediments have been performed along the north and south shores of the Inner Clyde estuary, UK. Surface sediment data reveal significant spatial variation in Cr content and an association with major sediment characteristics and location within the estuary. Depth variation for Cr and other heavy metals cannot be explained by variation in major geochemical controls such as grain size and organic matter and highlights the impact of historical contamination on sediment quality. These elevated levels at depth may still have environmental impact through redox-reactivity, in association with iron and manganese. Sequential extraction of sediments and pore water analysis of Cr(VI) and Cr(III) provide detailed information on release potential from the sediments. The implication of Cr mobility for biota in the estuary has been assessed by the analysis of a common marine bivalve, Mytilus edulis (Blue Mussel) and a burrowing polychaete, Nereis diversicolor (rag worm) from a number of survey sites. Bioconcentration factors for Mytilus indicate that the weakly held portion of sediment Cr is available for uptake and in the case of Nereis, bioaccumulation appears to be inhibited by sediment organic matter. PMID- 12901166 TI - The effects on human health and hydrogeochemical characteristics of the Kirkgecit and Ozancik Hot Springs, Canakkale, Turkey. AB - This investigation was carried out to determine the hydrogeochemical characteristics of the Kirkgecit and Ozancik hot springs. The study areas are located northeast and southwest of the town of Can, Canakkale. During the investigation, geological maps of the hot springs and its surroundings were prepared, and hot waters and rock samples were collected from the study sites. The Paleogene-Neogene aged andesite, trachyandesite, andesitic tuff, silicified tuff and tuffites form the basement rocks in the Ozancik hot spring area. In the Kirkgecit hot spring area, there are Lower Triassic aged mica and quartz schists at the basement rocks. The unit is covered by limestones and marbles of the same age. They are overlain by Quaternary alluvial deposits. A chemical analysis of the Kirkgecit hot water indicates that it is rich in SO(4)2- (1200.2 mgL-1), Cl- (121.7 mgL-1), HCO3- (32.5 mg L-1), Na+ (494 mg L-1), K+ (30.2 mg L-1), Ca2+ (102 mg L-1), Mg2+ (15.2 mg L-1), and SiO2 (65.22 mg L-1). Chemical analysis of the Ozancik hot water indicates that it is rich in SO(4)2- (575 mg L-1), Cl- (193.2 mg L-1), HCO3- (98.5 mg L-1), Na+ (315 mg L-1), K+ (7.248 mg L-1), Ca2+ (103 mg L 1), Mg2+ (0.274 mg L-1), and SiO2 (43.20 mg L-1). The distribution of ions in the hot waters on the Schoeller diagram has an arrangement of r(Na(+) + K+) > rCa2+ > rMg2+ and r(SO(4)2-) > rCl- > r(HCO3-). In addition, the inclusion of Fe2+, Cu2+, Cr3+, Mn2+, Ni2+ and Hg2+ in the hot water samples indicates potential natural inorganic contamination. The water analysis carried out following the ICPMS-200 technique was evaluated according to the World Health Organisation and Turkish Standards. The use and the effects of the hot water on human health are also discussed in the paper. PMID- 12901167 TI - Aluminium and fluoride concentrations of three tea varieties growing at Lantau Island, Hong Kong. AB - The present project aims to investigate aluminium (Al) and fluoride (F) contents in teas (Camellia sinensis (L.) O. Kuntze). Three different commercial tea varieties: Assam variety and two China sub-varieties, a large leafed variety and small leafed variety, were collected in two tea gardens of Lantau Island tea plantation of Hong Kong. In general, high concentrations of Al and F were accumulated in the mature leaves (15.3 and of 2.07 g kg-1 respectively). Among the three varieties, 'the small leafed' variety exhibited the highest Al and F contents followed by the 'large leafed' variety whereas the Assam variety had the lowest Al and F concentrations in its tea bushes. Tea products from a plantation were also analysed and it was noted that black tea had higher Al and F concentrations than green tea. The amount of Al and F released into tea liquor was also tested and the results showed that higher concentrations of Al and F were released into tea liquor under repeated infusion method than continuous infusion method. PMID- 12901168 TI - Application of cluster analysis to the geochemistry zonation of the estuary waters in the Tinto and Odiel rivers (Huelva, Spain). AB - The combination of acid water from mines, industrial effluents and sea water plays a determining role in the evolutionary process of the chemical makeup of the water in the estuary of the Tinto and Odiel rivers. This estuary is in the southwest of the Iberian Peninsula and is one of the estuarine systems on the northwest coast of the Gulf of Cadiz. From the statistical treatment of data obtained by analyzing samples of water from this system, which is affected by industrial and mining pollution processes, we can see how the sampling points studied form two large groups depending on whether they receive tidal or fluvial influences. Fluvial input contributes acid water with high concentrations of heavy metal, whereas industrial effluents are responsible for the presence of phosphates, silica and other nutrients. The estuarine system of the Tinto and Odiel Rivers can be divided into three areas--the Tinto estuary, the Odiel estuary and the area of confluence--based on the physical--chemical characteristics of the water. PMID- 12901169 TI - Environmental risk assessment of metals contaminated soils at silvermines abandoned mine site, Co Tipperary, Ireland. AB - A centuries long history of mining and mineral processing has resulted in elevated Cd, Pb and Zn soil concentrations in the vicinity of the Silvermines abandoned mine site (AMS), Co. Tipperary, Ireland. A process for preliminary evaluation of environmental risk was developed and implemented. Potential pathways of metal compound transport and deposition were mapped and used to optimise the subsequent site investigation. Elevated soil metals are shown to be predominantly in areas where metal deposition in soil is associated with water related pathways (surface runoff, seasonal groundwater seepage and floodplains). Extensive areas of soil in the surrounding district are classified as contaminated on the basis of Cd, Pb and Zn concentrations, both total and potential bioavailable (EDTA-extractable). The most affected areas, with metal concentrations in soil comparable with that within the AMS, were floodplains located 2-3 km downstream from the site. Assessment of the sequential effects on grass and grazing animals indicates that Pb poses the greatest risk due to its high toxicity and high concentrations in soil (more than 10,000 mg kg-1). Within floodplain areas grazing cattle may intake a lethal dose of Pb. On the basis of the investigation an approach to risk assessment was developed which allowed quantified assessment of the risks related to individual metals, areas of contamination and contamination targets. PMID- 12901170 TI - Environmental Se-Mo-B deficiency and its possible effects on crops and Keshan Beck disease (KBD) in the Chousang area, Yao County, Shaanxi Province, China. AB - Causes of Keshan-Beck disease (KBD) are still being probed and monitored in China. Relationships between trace elements from eco-environmental systems and KBD are poorly understood although relationships between environmental Se and human health have received extensive attention. In order to investigate relationships between eco-environmental geochemistry and KBD, we selected the Chousang KBD area in Yao County, Shaanxi Province, China, as an example of a prevailing KBD area applying I-Se-rich salts instead of utilizing Se-rich fertilisers on food crops to prevent local residents from developing KBD before 1995. Environmentally geochemical samples (rocks, soils, plants and children's hair) were collected from the Chousang KBD area. Soils in the study area contain 0.11 +/- 0.02 microgram Se g-1, 0.75 +/- 0.11 microgram Mo g-1, and 34.5 +/- 1.5 micrograms B g-1 on average, indicating that the study area is a deficient-Se-Mo B area. Se (0.07 +/- 0.007 microgram g-1), Mo (0.35 +/- 0.09 microgram g-1) and B (3 +/- 0 micrograms g-1) contents are low in wheat and corn used as a daily main food staple of local inhabitants. It is indicated that the study area is deficient in environmental Se-Mo-B for the local residents. Se contents of children's hair from the Yangyuan Elementary School in the study area range from 0.09 to 0.26 microgram Se g-1 with an average of 0.165 +/- 0.05 microgram Se g-1 (n = 10) in this KBD endemic area. Due to the low levels of Se, Mo and B available in soils and rocks, crops including wheat and corn are deficient in these elements, accordingly, the deficiency of Se, Mo and B in this area may be linked to the daily consumption of wheat and corn deficient in Se, Mo and B. Therefore, local inhabitants should be encouraged to fertilise mixtures of Se, Mo and B on crop plants in order to avoid development of KBD and guarantee a good harvest of crops. PMID- 12901171 TI - A modest proposal: (re)defining unconscious fantasy. AB - The author argues that unconscious fantasy, properly defined, necessarily represents the three-dimensional intersection of wishful thinking (fantasy), veridical perception of the environment (reality), and the naive cognition of childhood. It is proposed that, although attachment theory developed out of the intent to capture the unalloyed reality of dyadic experience, that experience is inextricably entangled with the other two components, wishes and naive cognition, and furthermore, that the behavior of children in the attachment paradigm can only be accounted for by positing the existence of underlying unconscious fantasies. In making these arguments, the author also addresses the development of unconscious fantasies and their relationship to compromise formations and trauma. PMID- 12901172 TI - The analyst's caseload as a family: transferences to fellow patients. AB - This article draws attention to a topic that has been mostly ignored in our literature: the manifold transference meanings of our patients' reactions to our other patients. Central aspects of the patient's transference may be displaced onto the analyst's other patients. The author includes clinical examples of hospitalized patients, where such transferences to other patients are often more dramatic and obvious, as well as examples from neurotic patients, where such transferences may be less dramatic and may tend to resolve more readily through interpretation. The displacement of transference feelings onto the analyst's other patients also has important implications for training analyses, where analysands are likely to have multiple relationships with the analyst's other analysands, supervisees, and students. PMID- 12901173 TI - Action knowledge, acknowledgment, and interpretive action in work with Holocaust survivors. AB - Survivors withhold disclosure of suffering when their terror is unwitnessed and when their expectation of disbelief or disregard obfuscates the reality of persecution. Knowledge itself then becomes traumatized, losing the power to inform and mobilize action. Survivors become habituated to suffering in a manner that subverts meaning, dampens vitality as well as pain, and arrests empathic connectedness. The dearth of transferential cues in such depleted existences leaves analysts in doubt as to whether they have been unintrusive or unavailable to these patients. Restoring survivors' sense of being witnessed requires interpretive actions that acknowledge the suffering that survivors have lost the will and means to make known or even represent. Such interventions draw on analysts' own projective identifications and use of the self, counterposing the will to live against the resignation to unwitnessed terror. PMID- 12901174 TI - A crucible for murder: the social context of violent children and adolescents. AB - The author describes aspects of social context that are of importance in the genesis of murderous violence in children and adolescents. Case presentations illustrate the effects of family attachment pathology, differing outcomes in destructive children, and the copycat phenomenon. The author concludes by summarizing the continuum of responses of children and adolescents to various forms of threat: real attack, unconscious threat, kindled threat, suppressed threat, bully-victim-bystander sadomasochistic ritual attack, and peer-group validated threat. PMID- 12901175 TI - Introduction to Michel de M'Uzan's "Slaves of quantity". PMID- 12901176 TI - Slaves of quantity. PMID- 12901177 TI - On being a dead, beloved child. PMID- 12901178 TI - [Regional differences of mortality from malignancies in Hungary]. AB - INTRODUCTION: The mortality of the Hungarian population is very unfavourable in relation to other European countries. Mortality from malignant diseases is the second most frequent cause of death in both sexes. The most frequent localisation of cancer is that of the bronchi and the lungs, followed by colorectal, breast and oral cavity cancers. AIM: Of the publication was to demonstrate the spatial distribution of mortality from malignant diseases of all sites, bronchi and the lungs, as well as mortality from cancer of the thyroid gland and leukaemias, and to evaluate the possible impact of the Chernobyl nuclear accident on the frequency of cancer mortality. METHOD: The spatial distribution of mortality in the country is evaluated by computing standardized mortality ratio on settlement level, using geographical information system. In case of frequent mortality events a region analysis was carried out, in the opposite case--a cluster analysis. RESULTS: Regarding the spatial distribution of mortality from all malignant diseases of 0-64 year-old males there are regions with excess mortality in almost each county. In case of women of this age group, there is a significantly higher mortality in Budapest, in three counties in the Eastern part of the country, and in some settlements in Transdanubia. Mortality from the cancer of the bronchi and the lungs significantly accumulates in both sexes in four counties in Eastern Hungary. Mortality from cancer of the thyroid gland and leukaemias does not show typical spatial accumulation as well. CONCLUSIONS: The premature mortality from all malignant diseases and of cancer of the bronchi and lungs of the Hungarian male and female population shows an increasing tendency. Mortality from the latter cause shows a typical spatial accumulation, which causes should be investigated in analytical epidemiological studies. The potential causative role of the Chernobyl accident could not be proven in any case. PMID- 12901179 TI - [Malignant vasovagal syncope]. AB - The diagnosis of epilepsy is complicated by various conditions that can mimic an epileptic seizure. Many patients with abnormal seizure activity during loss of consciousness may have cardiovascular syncope with global cerebral hypoxia (convulsive syncope), which may be difficult to differentiate from epilepsy on clinical grounds. The differentiation is, however, important because they need quite different treatment modalities. In addition, long-term anticonvulsant therapy is expensive and can cause serious morbidity. The authors present a case of a patient thought to have treatment-resistant epilepsy for years with recurrent seizure-attacks, who were subsequently found to have a malignant vasovagal reaction of 24s-asystole as a cause for the so called convulsive syncope. A simple, non-invasive evaluation of circulatory responses to acute orthostasis, the head-up tilt table test, can identify cardiovascular reflex abnormalities in patients with recurrent idiopathic seizure-like episodes. The authors could also reproduce the symptoms of the spontaneous attacks in their patient by this way, to confirm an alternative diagnosis of malignant vasovagal reaction and convulsive syncope in this patient with "refractory epilepsy". This rare cardioinhibition can be safely treated by dual-chamber pacemaker implantation, alleviating for the convulsive attacks, this therapeutic option was advised to this patient as well. Orthostatic stress tests should be considered early in the diagnostic workup of patients with convulsive blackouts. Cardiac causes of loss of consciousness should be considered in patients with presumed epilepsy, atypical premonitory symptoms, non-diagnostic electroencephalograms and failure to respond to anticonvulsant therapy. PMID- 12901180 TI - [Successful idebenone therapy of hypertrophic cardiomyopathy in Friedreich ataxia]. AB - Severe hypertrophic cardiomyopathy in a 15 years old child with Friedreich ataxia was treated with idebenone on the basis of a preliminary study reported in the literature. After 3 months of treatment the muscle thickness and mass and idices of diastolic function on echocardiogram and ischaemic signs on ECG changed significantly. IN CONCLUSION: Our data proves the preliminary results, so idebenobe treatment is very effective in the disease, where a deficiency of frataxin is involved in the regulation of mitochondral iron content which is responsible for myocardial injury. We suggest the widespread use of idebenone to treat patients with Friedreich ataxia and hypertrophic cardiomyopathy to improve the fatal prognosis of this type of cardiomyopathy. PMID- 12901181 TI - [Clinical characteristics of juvenile dermatomyositis]. AB - INTRODUCTION: Dermatomyositis, belonging to the group of the idiopathic inflammatory myopathies, is characterized by bimodal pattern of age-specific incidence of rates, with peaks in age group from 5 to 14 years (juvenile dermatomyositis) and in age group from 45 to 64 years (adult dermatomyositis). THE AIM OF THIS STUDY: Is to evaluate the clinical characteristics of 12 patients with juvenile dermatomyositis followed by the 3rd Department of Internal Medicine, University of Debrecen and the 2nd Department of Pediatrics, Semmelweis University, Budapest. METHODS: The authors analyzed the medical records of the patients with juvenile and adult form of dermatomyositis retrospectively. RESULTS: All of the children had symmetrical weakness of the proximal muscles. The most frequent cutaneous features were facial erythema and Gottron papules (11/12). The extramuscular manifestations were also assessed. 7 children had arthralgia. There were observed pulmonary fibrosis, Raynaud-syndrome, dysphagia and sicca-syndrome in the same patient, whose disease is overlapped with progressive systemic sclerosis. In view of the clinical course, the authors found that prevalence of polycyclic (relapsing-remitting) and monophasic subtypes of the disease were similar (6/12 and 5/12). Finally, all of the patients achieved remission, however, 2 patients have to take low-dose corticosteroid therapy permanently to maintain remission. One patient's cutaneous symptoms proved to be persistent and in further 2 cases, regression of the calcinosis is slow, but continuous. DISCUSSION: The authors compare their data of juvenile patients with the data of the relevant literature and to their experience with the management of adult DM patients. It seems to be reasonable to treat the patients in centres. PMID- 12901182 TI - [Determination of the lamivudine-resistant mutants of hepatitis B virus]. AB - INTRODUCTION: In addition to interferon, lamivudine is the other widely used antiviral agent in the therapy of chronic hepatitis B. This nucleoside analogue inhibits the RNA-dependent DNA polimerase and the reverse transcription by integrating in the viral DNA, which results in the secondary suppression of viral protein synthesis and replication of HBV. It has numerous advantages such as effective viral inhibition, mild side effects and the possibility of oral administration; on the other hand it poses the problem of time-correlated appearance of lamivudine resistant mutants during therapy. AIMS: In the Virusserology Laboratory of the Department I. Internal Medicine, Szent Gyorgy Hospital, Szekesfehervar, detection and type determination of the therapy resistant mutants in the C and B domains of HBV DNA polimerase gene has been carried out the for one year. In this paper, the authors review the molecular biological background of lamivudine resistance and summarise the applied test methodologies and the early results. PATIENTS: Six-month and/or 12-, 18-month samples of 18 chronic hepatitis B patients (4 women/14 men) treated in seven Hepatology Centres in Hungary were analysed. METHODOLOGY: Mutants of codons 528, 552, and 555 in the HBV polimerase gene were determined by nested polimerase chain reaction and reverse hybridisation. RESULTS: M528, V552, I552 and I555 mutants in different variations could be detected in ten out of 18 patients. CONCLUSIONS: Nowadays, drug therapy is the only treatment option used for the therapy of early and progressed chronic hepatitis B in Hungary. This new diagnostic technique was introduced to clarify the background of ineffective lamivudine therapy. Therapy resistance can occur due to the lack of reaction or the appearance of the special, therapy resistant mutants of the virus. Detection of these YMDD mutants together with the clinical picture and the biochemical and virological parameters can help in forming a decision about cessation of lamivudine therapy or application of a new drug. PMID- 12901183 TI - [Contribution to the problem of toxic shock syndrome]. PMID- 12901184 TI - ["Annoyance" and "rage", both reveal narrow mindedness...(On past and present of health care facility organization in the service of patient care)]. PMID- 12901185 TI - [Introduction]. PMID- 12901186 TI - [Thrombotic microangiopathy]. PMID- 12901187 TI - [Mechanisms of intraglomerular coagulation ]. PMID- 12901188 TI - [Mechanisms of endothelial damage in thrombotic microangiopathy]. PMID- 12901189 TI - Intravital videomicroscopy in peritoneal dialysis research. PMID- 12901190 TI - Molecular mechanisms involved in the peritoneal membrane exposed to peritoneal dialysis. PMID- 12901191 TI - [Early diagnosis, prevention and treatment of the peritoneal sclerosis syndromes]. PMID- 12901192 TI - [CA++/ calcineurin/NFAT signaling in endothelial activation and angiogenesis: effects od cyclosporin A ]. PMID- 12901193 TI - [Mechanism of inhibition of tumoral angiogenesis by thrombospondin-1]. PMID- 12901194 TI - [Role of VEGF in the cellular response to injury]. PMID- 12901195 TI - [Ascites and vascular dysfunction in human liver cirrhosis: new pathophysiologic concepts for a severe complication]. PMID- 12901196 TI - Antidepressants in general practice. PMID- 12901197 TI - Is primary care research a lost cause? PMID- 12901198 TI - Management of muscle and tendon injuries in footballers. AB - BACKGROUND: Muscle and tendon injuries are among the commonest football injuries and can be very frustrating to treat because of the high risk of recurrence. OBJECTIVE: This article aims to highlight the optimal management of, and risk factor for re-injury of, the common soft tissue injuries in football players. DISCUSSION: The high risk of recurrence of soft tissue injuries is because most football players return to play before complete healing of the injury. For hamstring and quadriceps strains and thigh haematomas, early return is often a sensible strategy. Calf strains in older players and tendinopathies (Achilles, patellar tendon and chronic groin injuries) generally require a more conservative approach as the risk of prolonged missed playing time is greater. The rupture of tendons that have nearby agonists can often be managed conservatively, but rupture of isolated prime mover tendons, such as the Achilles and patellar tendons, should be managed surgically. PMID- 12901199 TI - Preparation for skiing and snowboarding. AB - BACKGROUND: Winter sports participation is growing in popularity following the success of recent Australian Winter Olympic Teams. On the recreational front, snowboarding is equal in popularity to skiing. Injury types are varied and specific to the sports. OBJECTIVE: This article compares the common injuries sustained while skiing and snowboarding and details key factors that are assessed with our national mogul and snowboard teams. DISCUSSION: Before being able to provide advice regarding fitness and training as a preventive approach to reducing injuries, it is important to understand energy systems of winter sports. Suitable training regimens are considered and other preventive considerations, such as skill level, ski conditions and equipment are discussed with particular attention to specific energy system tests that can identify weakness. PMID- 12901200 TI - Snow skiing injuries. AB - BACKGROUND: Skiing is a sport that has exploded in popularity over the past 20 years. There are estimated to be 200 million skiers worldwide. OBJECTIVE: This article outlines the type and prevalence of ski injuries, and measures that have been made to reduce the incidence of injuries to skiers. DISCUSSION: The incidence of injury has decreased with the development of improved equipment, resort management and skier preparation. This decrease has been reflected in a lower rate of foot, ankle and tibial injuries. The overall incidence of knee injuries has increased and in particular an increase in more serious knee injuries. Snowboard injuries now account for approximately half the presentations to ski resort medical centres. PMID- 12901201 TI - Individualising therapy. The right dose, dosage form, frequency and duration. AB - BACKGROUND: Having chosen which drug to prescribe, the prescription now needs to be individualised for the patient in front of you. OBJECTIVE: This article discusses the factors that have to be considered when individualising drug therapy for each patient. DISCUSSION: Before considering writing a prescription it is important to develop a personalised approach for the individual patient. The prescription should also then be individualised as to the route of administration, dosage, dosage form, frequency, and duration of the medication being prescribed. PMID- 12901202 TI - GPs managing patients with eating disorders. A tiered approach. AB - BACKGROUND: General practitioners take on varying levels of responsibility for patients who have eating disorders. Roles appear to be tiered, from simply identifying illness and referring the patient on, to acting as care coordinator, providing medical and psychological treatment and/or continuing care. OBJECTIVE: Every GP has some level of responsibility toward this patient group. This article aims to identify the various levels of intervention and roles GPs may take and to provide practitioners with options regarding their level of clinical involvement. DISCUSSION: Just what role GPs choose to take in the management of patients with eating disorders depends on many factors including the stage of the patient's illness, and the doctor's interest, knowledge and clinical skills in this area of practice. PMID- 12901203 TI - Treatment of an earlobe keloid. Finding the evidence. PMID- 12901204 TI - Perils of pinna piercing and pseudomonas perichondritis. PMID- 12901205 TI - Mental health issues in Australian nursing homes. AB - BACKGROUND: Mental illness is common, under detected and often poorly managed in residential aged care facilities. These concerns have achieved greater prominence as the worldwide population ages. Over 80% of people in nursing home care fulfill criteria for one or more psychiatric disorders in an environment that often presents significant difficulties for assessment and treatment. OBJECTIVE: This article aims to provide an overview of the important mental health issues involved in providing medical care for patients with behavioural and psychological problems in residential aged care facilities. DISCUSSION: Recent developments in education and training, service development and assessment and treatment strategies show some promise of improving the outcome for aged care residents with mental health problems. This is of especial relevance for primary care physicians who continue to provide the bulk of medical care for this population. PMID- 12901206 TI - Haemorrhoids--a review. AB - BACKGROUND: Haemorrhoids have plagued humankind since the dawn of history. Hippocrates describes the treatment of anorectal disorders with special exactness. He describes specula to examine patients and he treated patients with suppositories, cautery and even by excision. It seems the fundamentals of haemorrhoid treatment have not changed for thousands of years. OBJECTIVE: This article presents a review of haemorrhoids with particular reference to the general practice setting. DISCUSSION: It has been estimated that 50% of the population has haemorrhoids by the age of 50 years. Although patients often consider the condition to be a single simple disease, this is not so. This article demonstrates that a large variety of closely related conditions are classified as haemorrhoids and it emphasises the dangers of misdiagnosis as haemorrhoids can mimic more important and serious diseases. PMID- 12901207 TI - Failure to diagnose--testicular torsion. PMID- 12901208 TI - Diagnosis and treatment of hand-arm vibration syndrome and its relationship to carpal tunnel syndrome. AB - BACKGROUND: Hand-arm vibration syndrome (HAVS) is a condition associated with the use of vibrating tools that occurs mainly in men. It consists primarily of 'occupational' Raynaud disease and digital polyneuropathy. Carpal tunnel syndrome (CTS) is also associated with hand transmitted vibration exposure and can coexist with HAVS. OBJECTIVE: This article examines recent papers on causation, diagnosis, relationship to CTS and treatment. A Medline search was conducted, as was a search of UK, USA and Australian government occupational health and safety websites. Published papers that were single case studies or of poor design were not included. DISCUSSION: There are no 'gold standard' diagnostic tests for HAVS. It can mimic CTS in temperate climates and can occur with CTS. This is the diagnostic challenge when a male worker presents with apparent CTS symptoms. If he has worked with vibrating tools for many years, a diagnosis of HAVS or co diagnosis of HAVS should be considered before a diagnosis of pure CTS is made. Nonwork risk factors for HAVS are predisposition, smoking, and exposure to vibration outside work. Cessation of exposure (and smoking) and redeployment is a critical part of treatment due to the dose response relationship of HAVS. This contrasts with adequately treated CTS, where the vast majority of workers can return to pre-injury duties. In severe cases, calcium antagonists are also used, but treatment is often ineffective. Few workplaces in Australia manage vibration risk or conduct screening to identify workers with early HAVS who should be redeployed. Local doctors have an important opportunity to diagnose HAVS and to make recommendations to the workplace on redeployment as part of treatment before symptoms become irreversible. PMID- 12901209 TI - Right lateral elbow pain. PMID- 12901211 TI - Does the evidence change your practice? Finding and using the best research evidence. PMID- 12901210 TI - Diabetes case study. Eye series 6. PMID- 12901212 TI - Teaching practical procedures in general practice. A primer for supervisors of medical students and registrars. AB - BACKGROUND: Practical procedures are central to many general practitioners' job satisfaction and their ability to provide comprehensive care. Despite this, students lack opportunities to learn skills and fewer GPs perform them. OBJECTIVE: This article develops a framework for teaching practical skills and their assessment in general practice. DISCUSSION: True competence means more than performing a task in isolation: knowledge, skills and attitudes are needed to perform in the workplace. Teaching should be closely related to assessment in general practice. A four stage method of instruction can facilitate learning of task skills. This should be followed by a series of case discussions, scenarios or simulations to teach and assess the learners' ability to manage tasks in the work environment. Linking teaching, assessment and feedback is integral to improving learning opportunities in general practice. PMID- 12901213 TI - Performance and remediation in general practice training. PMID- 12901215 TI - In search of justice. PMID- 12901214 TI - The ban on pregnant netballers. PMID- 12901216 TI - Grassroots research: how to have a stab at your first project and succeed. AB - BACKGROUND: Current initiatives are aimed at building the capacity within Australian primary care to undertake high quality research and evaluation, including clinicians who are not primarily researchers. OBJECTIVE: To provide a simple guide to encourage practice based research projects. TIPS AND ADVICE: Be parsimonious with the amount of data collected, ensure the literature is searched first, keep the project as simple as possible, invest time in planning and managing the project, readily ask for help and advice, and make every effort to publish the results. DISCUSSION: A modest, carefully supported and systematic approach is recommended for increasing the chance of success and quality of the research. PMID- 12901217 TI - Is there an increased role for general practice in making women more aware of folate supplementation to prevent neural tube defects? PMID- 12901218 TI - Painful nipples in nursing mothers: fungal or staphylococcal? A preliminary study. PMID- 12901219 TI - Diabetes clinical management guidelines. A self reported survey of GPs' awareness, attitudes and use. PMID- 12901220 TI - The influence of specialists on prescribing by GPs. A qualitative study. AB - OBJECTIVE: To investigate specialists' influence on prescribing by general practitioners. METHOD: A focus group study of 35 GPs. SETTING: Hunter Urban Division of General Practice, New South Wales. RESULTS: Although GPs thought specialists had only a small influence on their prescribing overall, it was substantial in some clinical areas, in complex conditions and conditions seen infrequently. Specialists were seen as authoritative and unbiased. Local specialists were particularly influential. Specialist influence came from seeing how specialists managed patients, clinical meetings, and specific verbal advice. It influenced the prescribing of new drugs, selection of drugs within a class and sometimes changed established prescribing practices. DISCUSSION: Interventions to change GPs' prescribing practices should address the importance of specialist influence, and not focus on GPs alone. PMID- 12901221 TI - The critical need for cancer biometrics: quantitative, reproducible measures of cancer to define response to therapy. PMID- 12901222 TI - Anti-estrogens in the treatment of breast cancer: current status and future directions. AB - For many years, the non-steroidal estrogen receptor (ER) blocker tamoxifen has been the drug of choice for the treatment of hormone-responsive breast cancer. However, deleterious effects on other tissues such as the endometrium and the frequent occurrence of relapse due to tamoxifen resistance have led to the search for alternative drugs that exhibit more favorable tissue-specific ER effects. Alongside these selective ER modulators, a new generation of aromatase inhibitors, which act by limiting the supply of local estrogen to the breast tumor, has emerged to challenge tamoxifen for supremacy. Together with other novel therapies under investigation, it can be hoped that such drugs may, alone or in combination, lead to better breast cancer treatment. PMID- 12901223 TI - Epidermal growth factor receptor inhibitors: an update on their development as cancer therapeutics. AB - Therapeutic agents targeting the epidermal growth factor receptor (EGFR) have recently been approved for use in patients based on the results of large-scale phase II studies involving patients with advanced refractory non-small-cell lung cancer (NSCLC). Disappointingly, results from phase III trials of gefitinib in combination with standard chemotherapy regimens for the treatment of NSCLC were negative. While results from phase III trials with other agents such as erlotinib and cetuximab will be reported in the next 12 to 18 months, the early results raise a number of questions regarding the development of these agents, including patient selection (e.g., disease, stage, prior therapy, EGFR or other biomarker expression) and combinations with standard treatment regimens as well as hormonal agents, radiation or other novel agents which will require further elucidation. Early data suggest a number of potential roles for these agents in the modulation of resistance and in combination with other inhibitors of signal transduction. PMID- 12901224 TI - Angiogenesis inhibitors in cancer therapy. AB - The formation of new blood vessels is essential for tumor growth and progression. Angiogenesis inhibitors have been demonstrated to block tumor growth and/or metastasis. Although initial clinical data have been disappointing, new strategies to increase the efficacy of these drugs have ensured their ongoing clinical development. This review highlights the broad spectrum of angiogenesis inhibitors under investigation in both preclinical and clinical studies. From 'natural' inhibitors to thalidomide analogs, the number of compounds in development is extensive. Elucidating the mechanisms of action will enable their use in conjunction with existing/other novel therapies, thereby maximizing the potential efficacy of angiogenesis inhibitors to fulfill their promise in patients with cancer. PMID- 12901225 TI - Telomerase inhibitors in cancer therapy: current status and future directions. AB - The ends of chromosomes (telomeres) are subject to progressive shortening in normal somatic cells, leading ultimately to irreversible growth arrest. In contrast, telomeres in all cancer cells are stabilized in length and effectively immortalized by the enzyme telomerase, which catalyzes the synthesis of telomeric DNA repeats. Several strategies have been devised for the inhibition of telomerase in the hope that this will result in anticancer effects. The principal approaches of catalytic inhibitors, antisense to the template, and folding of the DNA substrate, are reviewed and critically evaluated for their potential in anticancer therapy. PMID- 12901226 TI - CpG oligodeoxynucleotides for immune stimulation in cancer immunotherapy. AB - Synthetic oligodeoxynucleotides containing CG motifs (CpG ODNs) have potent immunostimulatory properties, and have potential as immunotherapeutic agents in cancer. Animal models suggest that CpG ODNs may be useful as both single agents and to enhance the efficacy of monoclonal antibody therapy. CpG ODNs can also enhance the efficacy of tumor immunization through use as an immune adjuvant, to activate antigen presenting cells, or as a component of DNA vaccines. Clinical trials are now beginning to assess whether the promise of these agents in the laboratory will translate into CpG ODNs becoming a valuable component of clinical cancer immunotherapy. PMID- 12901227 TI - Cancer therapeutic monoclonal antibodies targeting lymphocyte co-stimulatory pathways. AB - Monoclonal antibody (mAb) targeting to tumor antigen is an important therapeutic approach for the treatment of a variety of cancers. Both positive and negative co stimulatory pathways are critical components for the control and fine-tuning fo immune responses and recently mAb targeting of lymphocyte co-stimulatory pathways has been investigated. With the development of agonistic and antagonistic mAbs, both positive and negative pathways can be regulated and the potent therapeutic efficacy of this method has been demonstrated in experimental models. With high affinity and selectivity, as well as humanization technology for mAbs, there is little doubt that this new approach alone, or in combination with other therapeutic modalities, will play an important role in immunotherapy for cancers in the near future. PMID- 12901228 TI - Preclinical and clinical development of immunocytokines. AB - Advances in preclinical and clinical development have demonstrated that monoclonal antibodies and immuno-activating cytokines have a beneficial role in certain clinical oncology settings. Genetic engineering has now been used to create 'immunocytokines (ICs)'. These are fusion proteins that consist of an immune-activating cytokine linked to a tumor-reactive monoclonal antibody. Preclinical data demonstrate that ICs are far more effective in murine tumor models than the separate molecules from which they are derived. Clinical testing of ICs has recently begun using an anti-GD2 monoclonal antibody linked to interleukin-2 (IL-2) (hu14.18-IL-2), and using an antibody directed against the human epithelial cell adhesion molecule linked to IL-2 (KS-IL-2). PMID- 12901229 TI - Polymer-protein and polymer-drug conjugates in cancer therapy. AB - Polymer conjugates are becoming established as a new approach towards improved cancer therapy. These water-soluble, hybrid constructs fall into two main categories: polymer-protein conjugates (already available as licensed products), and polymer-drug conjugates (currently in clinical development). Polyethyleneglycol conjugation of proteins is accepted as a means to reduce immunogenicity, prolong plasma half-life and enhance protein stability. Polymer drug conjugation promotes tumor targeting by the 'enhanced permeation and retention' effect, and at the cellular level, allows lysosomotropic drug delivery. Eleven polymer-drug conjugates have entered clinical development and activity has already been observed in chemotherapy refractory patients. Certain compounds have also demonstrated a marked reduction in drug toxicity. PMID- 12901230 TI - Vaccinating against GD3 ganglioside using BEC2 anti-idiotypic monoclonal antibody. AB - GD3, a ganglioside expressed on tumors of neuroectodermal origin such as melanoma and small-cell lung carcinoma (SCLC), is an attractive vaccine target but is poorly immunogenic. Our group has pursued several strategies designed to immunize patients against GD3 including using BEC2, an anti-idiotypic monoclonal antibody that mimics GD3. Pilot trials in melanoma and SCLC indicate that BEC2 can induce an anti-GD3 antibody response in a subset of patients and also suggests that immunization with BEC2 is associated with improved survival. A phase III trial is underway in SCLC patients to determine the effects of BEC2 on overall survival. PMID- 12901231 TI - Developments in cancer vaccination. AB - The clinical application of cancer vaccines has progressed steadily from the rapid preclinical, technological advances of recent years. Cancer vaccine development has followed to some extent the pattern of vaccination for infectious disease, although features, such as the need to treat established disease in the face of immune evasion, vary. The necessity of conducting safety testing of new therapies in advanced cancer patients rather than in early-diagnosed patients often means that therapeutic benefit is seldom observed. Data that support even limited clinical efficacy are therefore encouraging. Initial approaches using whole irradiated tumor cells continue with some success, although there has been some refinement involving the use of tumor antigens and their subunits in a protein/peptide form (often via manipulated dendritic cells) or as DNA and RNA delivered naked or by viruses. Appropriate modes of boosting the immune system and immune conditioning appear important. Cancer vaccination research is likely to continue along many paths before any approaches become mainstream. PMID- 12901232 TI - Alpha v integrin inhibitors and cancer therapy. AB - Recent gene disruption experiments have suggested that targeting the alpha v integrins (cell surface adhesion and signaling receptors) to prevent tumor progression can result in different outcomes depending on the strategy. Nevertheless, two alpha v binding antagonists have made their way to the clinic in the oncology field; both Vitaxin, a humanized antibody, and cilengitide, a cyclic peptide mimicking the RGD ligand recognition peptidic domain common to alpha v integrin ligands, are in phase II clinical trials. This year, development of another peptidic inhibitor was initiated. This review questions whether companies are reluctant to propose small synthetic heterocyclic inhibitors as successors to peptide-derived agents with better pharmacokinetics and oral bioavailability. Is this class of compounds immediably flawed like the platelet alpha IIb beta 3 oral antagonists? Tentative answers are provided in this review following description of the lead compounds and the rationale for their use as cancer treatments, imaging agents or drug targeting vectors. PMID- 12901233 TI - BMS-184476 Bristol-Myers Squibb. AB - BMS-184476 is a taxane analog under development by Bristol-Myers Squibb for the potential treatment of solid tumors. By February 2001, it had entered phase II trials and, as of May 2002, phase II studies had been carried out in patients with a number of tumor types, including breast, ovarian and non-small-cell lung cancer. PMID- 12901234 TI - 109881 Aventis. AB - 109881 is an orally and intravenously active taxoid in development by Aventis Pharma (formerly Rhone-Poulenc Rorer) as a potential treatment for a variety of taxoid-resistant solid tumors, such as brain and lung cancers. As of April 2000, the compound was in phase IIa trials in the US, France, Belgium and Germany. Phase II trials were ongoing in May 2001, and the company was evaluating an oral formulation at this time. PMID- 12901235 TI - Cilengitide Merck. AB - Merck KGaA is developing cilengitide, the lead in a series of integrin antagonists with anti-angiogenic activities, for the potential treatment of a variety of cancer types. The National Cancer Institute is conducting clinical trials of cilengitide. In October 1999, phase II trials in non-small-cell lung cancer (as a monotherapy) and pancreatic cancer (in combination with gemcitabine) were initiated. These were ongoing in February 2002, by which time, a phase I trial and a phase I/II trial in glioblastoma were underway. PMID- 12901236 TI - BMS-247550 Bristol-Myers Squibb/GBF. AB - Bristol-Myers Squibb and the German Research Centre for Biotechnology are developing BMS-247550 as one of a series of epothilones, for the potential treatment of various forms of cancer. BMS-247550 had successfully completed phase I trials by September 2000 and by February 2001, phase II trials had been initiated for a variety of tumor types in collaboration with the National Cancer Institute; at this time, DNA filing was expected in 2003, and commercialization in 2004. By April 2002, phase I trials in children had also been initiated. As of May 2002, phase II studies had been conducted in patients with non-small-cell lung cancer and breast cancer. PMID- 12901238 TI - The surgeon at risk, or the hazards of practicing surgery. PMID- 12901237 TI - BAY-43-9006 Bayer/Onyx. AB - Bayer and Onyx are developing BAY-43-9006, an oral cytostatic Raf kinase inhibitor for the potential treatment of colorectal and breast cancers, hepatocellular carcinoma and non-small-cell lung cancer, in addition to acute myelogenous leukemia, myelodysplastic syndrome and other cancers. A US IND was filed in May 2000 and by February 2003 BAY-43-9006 was in phase II trials, with phase III trials expected to begin later in 2003. PMID- 12901239 TI - Relationship between caseload and morbidity and mortality in pediatric cardiac surgery--a four year experience. AB - BACKGROUND: The mortality rate associated with congenital heart surgery is apparently related to caseload. OBJECTIVE: To determine whether an increase in caseload over the long term at a single center affects management and outcome in children undergoing cardiac surgery. METHODS: Data were collected prospectively over a 4 year period from the computerized registry of the hospital's pediatric intensive care unit. Five parameters were analyzed: age at surgery, type of surgery, preventive measures (open chest), surgery-related and other complications (diaphragm paralysis and acute renal failure, respectively), and mortality. The data of a single-type surgery (arterial switch) were analyzed for bypass time and mechanical ventilation on an annual basis. RESULTS: The age distribution changed over the years, with more children under 1 year of age (20% newborns) undergoing surgery by the fourth year of the study. The caseload increased from 216 in the first year to 330 in the fourth, with a concomitant decrease in mortality rate from 4.9% to 3.2%. The chest was left open in 3.2% of patients in the first year and in 9.2% in the fourth year. The rate of diaphragm paralysis decreased from 6% to 2.4%. Death due to acute renal failure in patients requiring dialysis decreased from more than 80% in the first 2 years to 36% in the last two. These changes show an improvement but failed to reach statistical significance. Regarding the arterial switch operation, there was a significant improvement in pump time and duration of mechanical ventilation. CONCLUSIONS: The increase in caseload in pediatric cardiac surgery was accompanied by improved management, with a lower complications-related mortality rate. We suggest that for optimal care of children with congenital heart disorders, quality management resources should be concentrated in centers with high caseloads. PMID- 12901240 TI - Transthoracic echocardiographic assessment of proximal ascending aorta elasticity in familial heterozygous hypercholesterolemia patients. AB - BACKGROUND: Decreased elasticity of the aorta is associated with aging and several risk factors of atherosclerosis. The data regarding this phenomenon in patients with familial hypercholesterolemia are rather sparse. OBJECTIVES: To evaluate non-invasively the elasticity of the proximal ascending aorta of 51 heterozygous FH patients compared to 42 normal age and gender-matched controls. METHODS: Aortic elasticity was estimated by transthoracic echocardiography using the "pressure-strain" elastic modulus and aortic strain formulas. RESULTS: The elastic modulus score was higher in the FH group than in the controls (1.12 +/- 0.91 10(6) dynes/cm2 vs. 0.65 +/- 0.46 10(6) dynes/cm2 respectively, P = 0.01). This was consistent in both the pediatric (0.5 +/- 0.2 10(6) dynes/cm2 vs. 0.4 +/ 0.1 10(6) dynes/cm2 respectively, P = 0.009) and adult subgroups (1.3 +/- 1.0 10(6) dynes/cm2 vs. 0.8 +/- 0.5 10(6) dynes/cm2 respectively, P = 0.0004). Aortic strain was significantly lower in patients with FH than in controls (6 +/- 4% vs. 9 +/- 5% respectively, P = 0.0002). These findings reflected decreased elasticity of the proximal ascending aorta in the FH patients. In multivariate analysis, age, serum cholesterol level and serum triglycerides level were the independent predictors of the elastic modulus score, whereas age was the predictor of aortic strain. CONCLUSIONS: The elasticity of the proximal ascending aorta is decreased in heterozygous FH patients. PMID- 12901241 TI - Is the treatment of hyperlipidemia as secondary prevention adequate in different age groups in Israel? AB - BACKGROUND: Treatment of hyperlipidemia is important for secondary prevention in patients suffering from coronary heart disease. It has been proven that "young elderly" (patients aged 65-75 years) can benefit from the treatment at least as much as younger patients. OBJECTIVE: To assess the adequacy of treatment as part of secondary prevention in "young elderly" and younger patients. METHODS: In this prospective study, 389 patients discharged from the hospital with the diagnosis of coronary heart disease were divided according to age groups. Data were collected regarding lipid profile examinations, dietary and drug therapy, and results of lipid profiles. RESULTS: Less than one-third of the patients achieved target low density lipoprotein levels. More patients in the older age group achieved the treatment goals. The goals were achieved despite the fact that the percentage of patients treated with lipid-lowering drugs was lower in the older age group. CONCLUSION: The percentage of patients treated for hyperlipidemia as part of a secondary prevention plan in Israel is similar to that in other developed countries. The fact that more "young elderly" patients achieve adequate lipid profiles compared to younger patients indicates that there is no age discrimination towards this patient group. The finding that less than one-third of the patients reach the treatment goals should prompt physicians to treat hyperlipidemia more aggressively. PMID- 12901242 TI - Minimally invasive video-assisted mitral and aortic valve surgery--our initial clinical experience. AB - BACKGROUND: After the introduction of endoscopic techniques to other surgical fields like general surgery, gynecology and thoracic surgery, cardiac surgeons sought their own methods of using minimally invasive techniques. OBJECTIVES: To examine whether this approach is less invasive and yields better results, more desirable cosmetic results, and a more rapid and complete rehabilitation, maintaining safety, efficacy, and outcome equivalent to those of more established procedures, such as median sternotomy. METHODS: From January 2000 to July 2001, 22 patients underwent video-assisted port-access mitral or aortic valve repair or replacement with the Heartport system in our department, and one underwent closure of atrial septal defect. RESULTS: Intraoperative transesophageal echocardiography revealed excellent functional results. Total operating room time, perfusion time, and cross-clamp time with this technique decreased with our growing experience and remains stable. There were no intraoperative reversals to mid-steronomy, no mortalities, and only one complication 24 hours after surgery. CONCLUSIONS: Thoracoscopic assisted cardiac surgery (via port access) provides all the advantages of minimally invasive surgery, accelerates recovery, decreases pain, and maintains overall surgical efficacy, while avoiding the complications and pathology of mid-sternotomy. For appropriate patients, this is the method of choice in our department. PMID- 12901244 TI - Patterns of misdiagnosis of multiple sclerosis. AB - BACKGROUND: Multiple sclerosis is a chronic demyelinating disease of the central nervous system that presents with variable signs and symptoms. This variability in the clinical presentation may result in misdiagnosis, unnecessary referrals and misleading information to the patients. OBJECTIVES: To identify the types of misdiagnoses made on the presentation of MS. METHODS: Fifty consecutive MS patients were questioned on their early symptoms, their mental status, the disease course until the diagnosis was confirmed, and the different diagnoses they received. RESULTS: The patients had been referred to 2.2 +/- 1.3 specialists before seeing a neurologist, and learned about their disease 3.5 years after the onset of symptoms. Twenty-nine patients (58%) were initially given 41 wrong diagnoses. While the majority of women were misdiagnosed mentally, orthopedic work-up was offered to the men. Misdiagnosis of MS occurred most often in patients who presented with non-specific sensory symptoms that did not conform to a specific neurologic syndrome. The patients emphasized the fact that not knowing worsened their anxiety, whereas receiving the diagnosis enabled them to begin coping with their disease. CONCLUSIONS: MS is often overlooked when patients present with non-specific sensory complaints. The difference in type of misdiagnosis between men and women may reflect a gender-dependent bias in the way physicians interpret sensory complaints. PMID- 12901243 TI - Onchocerciasis among Ethiopian immigrants in Israel. AB - BACKGROUND: Onchocerciasis results from infestation by the nematode Onchocerca volvulus, and is characterized clinically by troublesome itching, skin lesions and eye manifestations. Since 1992, approximately 9,000 immigrants have arrived in Israel from the Kuwara province of northwest Ethiopia where the prevalence of onchocerciasis is particularly high. OBJECTIVES: To determine whether onchocerciasis is the cause of cutaneous and ocular symptoms among recent immigrants from the Kuwara province in Ethiopia. METHODS: We examined 1,200 recent immigrants from the Kuwara province residing at the Mevasseret Zion immigration center outside Jerusalem. Among them, patients with cutaneous signs suggestive of onchocerciasis underwent a skin-snip biopsy and a thorough eye examination. RESULTS: In the detailed skin examination performed in 83 patients, the most common skin finding was chronic papular onchodermatitis, found in more than 46 patients (55%); depigmentation and atrophy was found in 13 (15%) and 12 (14%), respectively. In 40 patients (48%), living microfilaria were detected in their skin snips. Of the 65 patients who underwent a through eye examination, 45 patients (66%) had ocular complaints. Corneal abnormalities were found in 55 of the 130 eyes (42%), active anterior segment intraocular inflammation and live microfilariae were found in 4 eyes (3%) and lens changes in 16 eyes (1%). Eleven eyes (9%) showed retinal or choroidal changes. CONCLUSIONS: Skin and eye manifestations associated with onchocerciasis are prevalent among symptomatic Ethiopians who immigrated to Israel from the Kuwara province. PMID- 12901245 TI - Cystic fibrosis in adults: a changing scene. AB - BACKGROUND: Cystic fibrosis is no longer a terminal illness of childhood and mean survival is now over 30 years. Adult patients with atypical CF are increasingly being diagnosed. In Israel, all patients are still followed in pediatric centers. OBJECTIVES: To describe our experience with adult CF, stressing the importance of adult-related health and psychosocial issues. METHODS: Twenty-five CF patients aged 20-50 years, constituting 44% of the 57 patients followed at our center, were analyzed for pulmonary and extrapulmonary features and management. RESULTS: Nineteen were diagnosed as children and 6 as adults. Nineteen were pancreatic insufficient and 6 were pancreatic-sufficient, including 5 diagnosed as adults. Pulmonary status was usually stable, with forced expiratory volume in 1 second 66.3 +/- 21% (mean +/- SD) and no difference between pancreatic-sufficient and insufficient patients. The latter had more hemoptysis, Pseudomonas infection, intestinal obstruction, liver disease and diabetes. Two patients died of malignancy and two of advanced lung disease. A majority received continuous inhaled and oral antibiotics, bronchodilators, Dnase, physiotherapy and periodic home intravenous antibiotics. Psychosocial functioning was excellent: 60% were employed, 36% were married and 40% had children (none with CF). Patients diagnosed as adults had mild multisystem disease or isolated severe lung disease. CONCLUSIONS: CF adults generally have a good quality of life. Advances in understanding the CF defect and a plethora of new treatment modalities bode well for the future. Patients must be maintained in optimal condition to reap the benefits, and there is an urgent necessity for adult physicians to develop expertise in CF. PMID- 12901246 TI - Radiofrequency ablation for the management of liver tumors. AB - BACKGROUND: Radiofrequency ablation has recently become a viable treatment option for unresectable primary or secondary lesions confined to the liver. OBJECTIVE: To study the local therapeutic efficacy, side effects and complications of radiofrequency ablation for the treatment of hepatocellular carcinoma and liver metastases. This is the first reported experience of radiofrequency ablation for treating malignant hepatic tumors in Israel. METHODS: Fifteen consecutive patients, aged 53-73 years, with 23 lesions (8 patients with HCC and 7 with secondary liver tumors) underwent radiofrequency ablation under general anesthesia. RITA nine-array 5 cm thermal ablation catheter and the model 1500 generator were used. The mean diameter of all tumors was 4.28 cm (range 1-10 cm). Three lesions were 1-3 cm in diameter (small), 17 lesions measured 3.1-5 cm (medium), and 3 measured 5.1-10 cm (large). RESULTS: Complete necrosis was found in 8 (66%) of 12 HCCs by computed tomography scan. Of the remainder, diffuse tumor recurrence was demonstrated in three lesions (25%) after lipiodol injection and there was one local tumor recurrence. In the metastases group complete necrosis was found in 5 of 11 lesions (45%). One major complication (peritonitis) was treated with antibiotics and four (26%) minor complications (right pleural effusion, small subcapsular hematoma) were monitored. CONCLUSIONS: Radiofrequency ablation appears to be an effective, safe and relatively simple procedure for the treatment of liver tumors. PMID- 12901247 TI - Lower extremity amputations in chronically dialysed patients: a 10 year study. AB - BACKGROUND: Lower limb critical ischemia is a major problem in dialysed patients. OBJECTIVE: To evaluate the results of revascularization procedures, amputations and prosthetic rehabilitation in dialysed amputees. METHODS: In this retrospective study we examined the charts of selected dialysis patients. Forty eight patients had undergone major amputation (4.5% of the dialysis population), and 24 patients entered the rehabilitation program. Widespread arterial calcification was common and led to falsely elevated ankle-brachial pressure indices in 9 of 14 limbs. Eight patients underwent revascularization. Subsequent major amputation was carried out 4 +/- 4.5 months after the revascularization (above knee in 5 patients and below knee in 3). Of the 16 patients who underwent primary amputation, only 2 were above-knee amputees. Seven patients with toe or metatarsal amputation went on to a major amputation 1.8 +/- 1.2 months after the distal amputation. RESULTS: No differences were found between diabetic and non diabetic patients regarding the number of revascularization operations performed, the level of major amputation, or overall survival. Prosthetic rehabilitation was considered successful in 12 patients, partially successful in 8, and failed in 4 patients. Patient survival time was shortest in those patients with failed rehabilitation. A younger age confirmed favorable rehabilitation results, while long-standing diabetics and bilateral amputees were poor rehabilitation candidates. Patients who underwent primary amputation had more successful rehabilitation. A comparison between 24 dialysed amputees and 138 non-uremic amputees revealed similar rehabilitation results, although hospitalization time was longer in the dialysed patients. CONCLUSIONS: Early definitive therapy is essential when dealing with critical ischemia. After diagnostic angiography, proximal revascularization should be performed where feasible. Primary amputation is indicated in patients with extensive foot infection or gangrene. Prosthetic rehabilitation is warranted in most dialysed amputees. PMID- 12901248 TI - Diffuse idiopathic skeletal hyperostosis: a distinct clinical entity. AB - Diffuse idiopathic skeletal hyperostosis is often incorporated into osteoarthritis. Although DISH often coexists with OA, patients affected by this disorder differ from patients with primary OA in several aspects: prevalence in the general population, gender distribution, anatomic site of primary involvement, magnitude and distribution in the spine and the peripheral joints. DISH is a distinct clinical entity. Its recognition as such should stimulate clinicians and researchers to focus on its pathogenesis, treatment and prevention. PMID- 12901249 TI - Osteoporosis in pediatrics. PMID- 12901250 TI - The coagulopathy of sepsis: pathophysiology and management. AB - Sepsis is an infection-induced inflammatory syndrome that results in a complex network of adaptive and maladaptive alterations in homeostatic mechanisms. Severe sepsis, defined as sepsis associated with acute organ failure, is a serious disease with a mortality rate of 30-50%. The coagulation system, through complex interactions, has an important role in the final outcome of the sepsis-induced inflammatory cascade. A fine and delicate balance that normally exists between anticoagulant mechanisms and the procoagulant response is altered in sepsis. Activated protein C, an endogenous vitamin K-dependent anticoagulant, plays a major role in the down-regulation of the procoagulant arm. It also possesses anti inflammatory properties. Endothelial damage during sepsis impairs the endothelium dependent activation of protein C, thus shifting the balance towards thrombosis. This shift may contribute to the development of sepsis-related multi-organ failure. Evidence suggesting that activation of the coagulation system may contribute to sepsis-related morbidity and mortality has led to extensive research attempting to correct the hemostatic defects seen in septic patients. Indeed, a recent randomized controlled trial demonstrated a reduction in overall mortality in patients with severe sepsis treated with APC. In this review we discuss the pathogenesis of the coagulopathy of sepsis, as well as the new therapeutic approaches aimed at correcting the defects in the coagulation system. PMID- 12901251 TI - Volume and outcome--time to step forward. PMID- 12901252 TI - Onchocerciasis (river blindness). PMID- 12901253 TI - Induced sputum as a diagnostic tactic in pulmonary diseases. AB - The induced sputum technique allows sampling of the airways in a non-invasive manner and thus offers a unique opportunity to identify biomarkers of potential clinical utility in respiratory medicine. Sputum cells were originally examined in stained smears and the procedure was applied in both research and clinical settings from the 1950s through the 1970s. The cells, recovered from spontaneous coughing, were used to study lung cancer and respiratory infections and, later on, to diagnose Pneumocystis carinii pneumonia in patients infected with human immunodeficiency virus. The method was largely improved by the induction of sputum with aerosol of hypertonic saline and was extended to become part of the assessment of airway inflammation in bronchial asthma and chronic obstructive pulmonary disease. It was recently shown that induced sputum can be used to study interstitial lung diseases and, more specifically, sarcoidosis, non-granulomatous ILD, occupational lung diseases and other systemic diseases with lung involvement. PMID- 12901254 TI - Total hip replacement via a combined intra and extra-pelvic approach causing dysuria and bladder penetration. PMID- 12901255 TI - Early prenatal diagnosis of conjoined cephalopagus twins. PMID- 12901256 TI - Strychnine intoxication in a child. PMID- 12901257 TI - Lymphangioleiomyomatosis: a rare indication for lung transplantation. PMID- 12901258 TI - Cobalamin deficiency in a breast-fed infant of a vegetarian mother. PMID- 12901259 TI - Evaluation of spermatogenesis using flow cytometry and confocal microscopy. PMID- 12901260 TI - [Cardiogenic shock in the course of myocardial infarction--the results of treatment during hospitalization and in long-term follow-up]. AB - Cardiogenic shock develops in 5-15% of patients hospitalised with acute myocardial infarction. It is responsible for more than a half of all hospital deaths with survival rate of about 20%. Conventional medical therapy with use of adrenergic, vasoactive, inotropic and thrombolytic agents has failed to improve survival. Treatment strategy combine hemodynamic stabilisation with restoration of coronary blood flow. The aim of the study was evaluation of mechanical restoration of coronary blood flow in infarction related artery and to assess its influence on mortality in patients with myocardial infarction complicated by cardiogenic shock. We retrospectively analysed 58 subjects: 26 patients treated by primary angioplasty, 25 patients with PTCA angioplasty after streptokinase treatment and 7 ones treated conservatively. TIMI 3 flow in angioplasty treated patients was achieved in 70.6% with in hospital mortality rate 14%, however, when reperfusion was unsuccessful the mortality was high (80%). 12 months follow-up mortality rate was 41.8%. CONCLUSION: Successful reperfusion with coronary angioplasty of the infarct-related artery can significantly reduce mortality rate in patients with cardiogenic shock. Patients who survived in-hospital period have favourable one-year prognosis. PMID- 12901261 TI - [Urinary system abnormalities in children of the pediatric outpatient clinics in Lublin]. AB - The study was carried out in 3160 children screened for urinary system abnormalities. The examined group consisted of patients of the Pediatric Outpatient Clinics in town Lublin, born between 1992-1996. The comprehensive clinical, anamnestic and laboratory analysis was based on the data from case records and hospital cards. The abnormalities of urinary tract were found in 112 children. It constituted 3.5% of the studied patients and 20.7% of children with urinary tract infections. It emerged that the urinary tract infections occurred in all children with urinary defects. The abnormalities predominated among girls than boys (60.7% and 39.3% respectively). Pyelocalycectasia and bilateral vesico ureteral reflux were the most often diagnosed malformations of urinary tract. PMID- 12901262 TI - [Clinical analysis of CAPD-treated patients without anemia]. AB - Very few patients with chronic renal failure (CRF) can maintain a hemoglobin concentration > 12 g/dl without rHuEpo treatment. Among 68 patients treated in our Department by CAPD, 12 of them (17.6%) with daily diuresis < 400 ml, for at least 6 months had Hb level over 12 g/dl, Hct > 35% without rHuEpo treatment. In present study we carried out a clinical analysis of 12 patients without anaemia treated CAPD from 12 to 63 months. Mean Hb value was 13.5 +/- 0.97 g/dl (range 12.4-14.9 g/dl). None of 12 patients had acquired cystic kidney disease in ultrasonographic investigation. Nobody of these patients was rHuEpo treated. Mean C-reactive protein as well as ferritin serum concentrations was significantly lower in this group of patients compared to others. Mean endogenous Epo concentration was not significantly different from other CAPD pts. We conclude that spontaneous increase in Hb concentration in CAPD pts was not connected with increase in the serum endogenous Epo concentration. Factors other than Epo probably play a role in regulation of erythropoiesis in these pts. PMID- 12901264 TI - [Subpopulations of CD4 and CD8 lymphocytes in peripheral blood of patients with non-toxic thyroid nodular goitre]. AB - The aim of the research was to establish the role, which is played by the immune system is in the pathogenesis of non-toxic thyroid nodular goitre. For this purpose the expression of phenotypic markers of T-helper (CD4) and T suppressor/cytotoxic (CD8) lymphocytes was estimated. The study was performed on 27 patients with histopathologically confirmed non-toxic thyroid nodular goitre. The control group consisted of 28 healthy blood donors. In patients with thyroid goitre the percentage of lymphocytes was significantly higher than in the control group (p < or = 0.0001). Likewise the percentage of T-helper (CD4) and T suppressor/cytotoxic (CD8) lymphocytes as well as the CD4/CD8 ratio was significantly higher in patients with thyroid goitre. The results of this study show that the immune system has important role in pathogenesis and development of non-toxic thyroid nodular goitre. PMID- 12901263 TI - The importance of exhaled air condensate in assessing the oxidant-antioxidant system in patients with chronic obstructive pulmonary disease. AB - The aim of this study was to investigate the relationship between selected lipid peroxidation products as markers of oxidative stress and antioxidant defense capacity in exhaled air condensate in comparison to blood and COPD severity. We detected that the increase of lipids peroxidation products in exhaled air condensate (less in blood) was accompanied by the reduction of total antioxidant capacity (antiradical activity and ceruloplasmin) according to the severity of COPD. Thus, the level of malondialdehyde significantly increased both in blood and exhaled air condensate in COPD1,2,3 (by 24%, 86%, 100% and 58%, 92%, 2,3 times respectively). Antiradical activity level was significantly decreased both in blood and exhaled air condensate in patients with COPD2,3 by 24%, 51% and 23.31% respectively. The results of this study clearly demonstrated the imbalance in oxidant-antioxidant system in advanced COPD. PMID- 12901265 TI - [Short saphenous vein incompetence as a cause of recurrent varicose veins]. AB - The aim of the study was the retrospective ultrasound Doppler evaluation the patients with recurrent varicose veins after surgery of the short saphenous vein incompetence. This study was performed in 35 patients (42 limbs). Mean interval between the time of operation and ultrasound Doppler evaluation was 4.9 years. The causes of the recurrence were divided due to F. Vin and F. Chleir classification. Recurrences were classified in five categories. The main causes of recurrence were: the remains of the short saphenous vein in primary operation (52.3%) and junctional stump with incontinent collaterals (26.1%). CONCLUSIONS: The recurrence of varicose veins is mostly associated with incompetence of short saphenous vein. The colour-coded duplex sonography is a modern tool in preoperative diagnostics of anatomical variety of short saphenous vein and in establishing the cause of the recurrence. USG evaluation could make precise operation easier and in a case of recurrence helps to eliminate the origin of reflux. PMID- 12901266 TI - Population dependent differences of distribution of congenital heart malformations. AB - The aim of the work was to study the pattern of congenital circulatory system malformations in the newborns in Upper Silesia, Poland (N = 355), and to compare it with the data known from the literature, predominantly with genetically very distant Chinese population from Hong Kong (N = 744). There were significantly more boys (about 60%) than girls (about 40%) in both groups. Much higher rate of the heart defects with pulmonary outflow obstruction was found in Hong Kong neonates (37.8%) than in the Polish ones (16.1%). Left ventricular outflow obstruction malformations were more common in Chinese group (22.7%) than in Polish one (14.1%), as well. On the other hand, Polish neonates were significantly more often admitted to cardiological hospital because of left-to right shunts (37.7%) as compared to Chinese population (15.5%). The rate of complete transposition of great arteries was similar in both studied groups (12.1% in Poland versus 12.4% in Hong Kong). So called common mixing malformations were not significantly more often found in Polish newborns (12.1%) than in Hong Kong ones (8.3%). It is concluded that it significant race dependent distribution difference of congenital heart malformations exists between Polish (Silesian) and Chinese (Hong Kong) newborn populations. On the other hand the pattern of congenital heart malformations found in Polish newborns seems to be not very different from these ones, known from literature, described in the USA or in Europe. PMID- 12901267 TI - [Treatment of incarcerated abdominal hernia]. AB - The aim of the study is to present problems connected with a diagnosis and treatment of incarcerated abdominal hernia. In the years 1990-2001, 142 patients with incarcerated abdominal hernia were admitted to hospital. In this group 112 patients were urgently operated on. These were 25 femoral hernias, 22 inguinal hernias, 23 umbilical hernias, 41 postoperative hernias and 1 internal hernia. RESULTS: The patients presented on clinical picture gastrointestinal tract obstruction or peritonitis. Postoperative mortality was 5.4%, however, in the group of patients with incarcerated postoperative hernias 17.5%. CONCLUSIONS: Treatment of incarcerated abdominal hernia is a serious surgical problem. Operations are marked by high mortality due to the late diagnosis of incarceration and further postoperative complications. PMID- 12901268 TI - [Acute normovolaemic haemodilution in children]. AB - Acute normovolemic haemodilution was performed in 34 children (4-6 years old, weight: 16-68 kg) in whom major blood loss was expected during operation. Anaesthesia was induced with thiopentone, and then maintained with N2O/O2, fentanyl and pipecuronium. IPPV was used to maintain normocapnia. Heart rate, blood pressure, central venous pressure and core body temperature were monitored during haemodilution and throughout the surgery. A predetermined volume of blood to reduce hematocrit value to 30% was withdrawn in standard collection bags. In one child from Jehovah's Witness family hematocrit following haemodilution was only 20%. Every 1 ml of withdrawn blood was replaced with 1 ml of Haemaccel (Boehring, 32 children) or with 3 ml of isotonic crystalloid solution (PWE Polfa, 2 children). The blood lost during surgery was replaced with crystalloid solution and Haemaccel. Normovolemic loss of blood to haematocrit value of 20-24% was permitted. Retransfusion of collected blood was started when major bleeding was over, or when hematocrit decreased below 20-24%. Only 8 children needed homologous blood transfusion. Heart rate decreased significantly during haemodilution, while blood pressure remained stable. Low initial haematocrit and coagulopathy due to dilution of the plasma clotting factors were the limiting factors of the method. Partial thromboplastin time and plasma fibrinogen should be monitored throughout surgery, because the occurrence of coagulopathy promptly allows to initiate retransfusion of autologous blood. PMID- 12901269 TI - [The application of cryogenic temperatures in medicine]. AB - An increased interest in effects of low temperatures on human organisms has been observed during last years. Low temperatures have been applied in cryotherapy and cryosurgery. In our article we show the influence of low temperatures on different physiological mechanisms and the possibility of their application in the therapy. PMID- 12901270 TI - [Cholangiocarcinoma--bile ducts cancer]. AB - Cholangiocarcinoma (CC) is a malignant neoplasm deriving from intra- and extrahepatic bile ducts. It affects both sexes, and is most prevalent at the age 50 to 70. Chronic nonspecific ulcerative colitis, primary sclerosing cholangitis, hepatolithiasis, congenital hepatic fibrosis, and Caroli's disease may lead to the increased incidence of CC. Recently, hepatic cirrhosis in the course of virus associated chronic hepatitis has been suggested to be involved in the pathogenesis CC. Histologically, 90-95% of CC are well differentiated adenocarcinomas. Usually the tumor grows slowly and metastazes late locally and even less frequently extrahepaticly. CC often causes symptoms by blocking the bile ducts, abdominal pain, weight loss, signs of portal hypertension, rare ascites and thrombophlebitis. Serum chemistry was compatible with obstructive jaundice. The increased expression of CEA, Ca19-9, as well as loss or reduction of sialomucin/sulfomucin concentration in the biliary lining epithelium may be indicative of malignant changes. CC as usually non-vascularized nonencapsulated tumor with a large amount of fibrosis. It is isochogenic in classical USG, CT or MRI. MRCP-magnetic resonance cholangiopancreatography and virtual endoscopy are more helpful methods on the diagnostics of CC. Recently, FDG positron emission tomography has been suggested to be a sensitive technique in identifying small bile duct cancers. Surgical excision of the lesion confirmed localized CC. The adjuvant radio- and chemotherapy and transplantation are not satisfactory. Palliative therapy includes surgical biliary-intestinal bypass procedures as well as operative and nonoperative techniques for biliary intestinal drainage. Recently, the local treatment of CC by photodynamic therapy as a palliative strategy is very promising. Ordinary CC is reported as a neoplasm with a poor prognosis. Post resection 5-year survival is affirmed in about 25% of CC, whereas after palliative treatment only 1 year. PMID- 12901271 TI - [Alcoholic liver disease]. AB - Ethanol toxicity on liver is a function of duration of alcoholism, amount of daily intake of alcohol and patient's nutrition. The threshold of alcohol toxicity on the liver is about 40 g of ethanol daily in men and 20-30 g in women, however liver cirrhosis develops in no more than 8-20% of patients exceeding this values. Ethanol is oxidized in the liver to acetaldehyde--a compound considerably more toxic than ethanol itself. Despite small amount of alcohol dehydrogenase (ADH) found in gastric mucosa, the metabolism of ethanol in this site may have an important hepatoprotective effect. The oxidation of ethanol is associated with a change of hepatocyte redox homeostasis, which leads to a number of metabolic disorders such as lactic acidosis, hyperlipidaemia and hyperuricaemia. Chronic ethanol consumption does not influence ADH activity, but has a profound stimulatory effect on microsomal enzymes, in particular cytochrome CYP2E1. This fact is responsible for development in alcoholic liver associated with rise of oxygen consumption, excessive production of free radicals and increased metabolism of ethanol, vitamin A and testosterone. Ethanol and acetaldehyde have a deleterious effect, both the direct and indirect, on hepatocytes e.g., generating radical oxygen species and damaging intestinal mucosal barrier. Cellular oxidative stress that is caused by both an excess of free radicals and the antioxidatives' deficiency (glutathion, vitamin E, phosphatidylcholine), may be the principal factor responsible for progression of alcoholic liver disease. Among other factors accelerating alcohol-related liver lesion there are certain drugs, high fat diet, infection with HCV and genetic factors (female sex, enzymatic polymorphic forms of ADH and ALDH, hemochromatosis). Great importance in pathogenesis of necrotic and inflammatory hepatic events is being attributed to portal endotoxaemia and cytokines induced within the liver, in particular TNF alpha and interleukin 8. These cytokines play a key role in development of alcoholic hepatitis, which clinical severity ranges from subclinical to fatal forms. Apart from abstinence, the treatment of alcohol liver disease is based on hyperalimentation, since alcoholism is generally associated with protein malnutrition. In severe forms of alcohol hepatitis corticosteroids are recommended. PMID- 12901272 TI - [Prognostic difficulties in pseudomyxoma peritonei]. AB - Pseudomyxoma peritonei is a rare disease leading to the accumulation of gelatinous masses in peritoneal cavity, usually secondary to neoplasmatic process of appendix or ovary. The authors describe a case report of 50 years old man with pseudomyxoma peritonei. Literature based discussion on prognostic difficulties and operative therapy perspectives in patients with this disease is presented. PMID- 12901273 TI - [Familial adenomatous polyposis in 48-year-old woman]. AB - In this study we presented the case of 48 years old woman who underwent surgery in the hospital in Poznan because of familial adenomatous polyposis. The disease was diagnosed in 1995, but then due to the fear of the surgery the patient did not consent to have an operation. We emphasized how it is important to motivate patients and explain them the need of surgical treatment in familial adenomatous polyposis. PMID- 12901274 TI - [Difficulties in the diagnosis of thyroid cancer in 8-year-old boy]. AB - The 8-year-old boy with 13 months history of enlargement of right cervical lymph nodes is described in this paper. Infectious lymphadenopathy was initially diagnosed. He was treated with antibiotics without success. Fine-needle aspiration biopsy in one of the enlarged lymph nodes showed the cells of normal thyroid gland without any signs of atypical growth. USG and scintigraphy revealed normal thyroid gland without nodules; no extra thyroid glands were detected. In 13th month of the disease two enlarged (about 5 ml each), painful, rather hard cervical nodules were present. The thyroid gland was no enlarged and without any palpable nodules. Chest X-ray and second thyroid scintigraphy were normal. Ultrasound imaging of thyroid gland revealed hypoechogenic lesion 11 x 9 x 12 in diameter in the upper part of right thyroid lobe. The morphological findings of removed nodules allowed to make a diagnosis of carcinoma papillare psammomatosum glandulae thyroideae. Total thyroidectomy with removal of cervical ipsilateral lymph nodes was performed. The operation is to be followed by radioactive iodine ablation and thyroid hormone suppression. The authors analyzed the causes and difficulties in diagnosis. PMID- 12901275 TI - [Two cases of strangulated obturator hernia]. AB - Two cases of strangulated obturator hernia were presented. Both cases were diagnosed during surgical intervention. In one case the postoperative recovery was not successful--mostly because of delay in referring to the surgeon. Described cases confirm diagnostic difficulties associated with this rare type of hernia. The attention should be paid to association of obstructive symptoms with obturator neuralgia and/or painful mass in rectal examination. PMID- 12901276 TI - [Drug expenditures of pensioners in 1997-2000]. AB - The general purpose of the study, carried out in the group of pensioners was to determine the relation between drug prices, household income and amounts of money spent on drugs in the years 1997-2000. The study was based on representative data gathered from annual household budgets review by Polish Statistical Office and data from pharmaceutical market published in 'Vitamina C++' magazine. The used method combined descriptive, comparative, table-descriptive analysis with graphical analysis. During studied period the real value of expenses on drugs in pensioners' households rose by 39.3% and available income decreased by 5.8%. Increased expenses on drugs caused the rise of the proportion of on spending on drugs in total household expenditure. It rose from 3.9% in 1997 to 5.2% in 2000. Throughout this time period the drug prices increased in real terms: the highest growth (approx. 49%) was noticed in patients' co-payment to reimbursed drugs. Despite rise in spending on drugs, due to the increase in drug retail prices and increasing patients co-payment, pensioners in comparison with 1997, could buy only approx. 93% units of reimbursed drugs in 2000. The possibility of buying drugs within OTC group increased by 18%. PMID- 12901277 TI - [Medical deontology--historical study]. AB - The subject of the paper was to present selected publications concerning the medical deontology. Special attention was paid on three of them. Well-known publications Hippocrates' oath formulated in 5th/4th century BC, Majmonides' prayer (12th century) and Polish medical deontology code published in 1994 underwent a comparative analysis. The objective of the analysis was the description of the similarities and differences in the assumptions constituting in the fundamentals of medical deontology. Its formulated in almost one thousand year intervals, as well as assumptions comparison of Polish and universal medical deontology. PMID- 12901278 TI - Prognostic significance of the deleted in colorectal cancer gene protein expression in high-risk resected gastric carcinoma. AB - The deleted in colorectal cancer (DCC) gene is a candidate tumor suppressor gene that may be associated with differentiation and proliferation of normal cells. Loss of heterozygosity (LOH) of 18q, where the gene is located, and absence of DCC protein expression have been associated with worse prognosis in certain subgroups of patients with colorectal adenocarcinoma. We studied the prognostic significance of loss-of-protein expression in 66 patients with resected gastric cancer with a high probability of relapse (T3, T4, N+). The DCC protein was detected with immunohistochemistry using an anti-DCC monoclonal antibody on paraffin-embedded sections. The DCC protein expression was present in 51 cases (77.3%) and absent in 15 cases (22.7%). Poorly differentiated and signet ring carcinomas had significantly lower expression than more differentiated tumors (p < 0.05) as did diffuse-type tumors compared to intestinal and mixed (p < 0.01). There was no correlation with proliferation rate, estimated immunohistochemically using an anti-proliferating cell nuclear antigen (PCNA) monoclonal antibody. Absence of DCC protein was an independent favorable prognostic factor (median survival 57 months vs. 18 months, p = 0.0176). The DCC protein expression was correlated with relapse site: all patients with distant metastases were positive for DCC staining, while one-third of patients with local/peritoneal relapse were negative (p < 0.01). In conclusion, DCC protein expression seems to be a significant prognostic factor in high-risk resected gastric cancer. Our results support previous data associating the DCC gene with differentiation and indicate that this gene may play a role in the metastatic potential of these tumors. These findings need to be confirmed by future larger studies. PMID- 12901279 TI - Immunotherapy with autologous, human dendritic cells transfected with carcinoembryonic antigen mRNA. AB - Immunizations with dendritic cells (DC) transfected with RNA encoding tumor antigens induce potent tumor antigen-specific immune responses in vitro and in murine models. We performed a phase I study of patients with advanced carcinoembryonic antigen (CEA)-expressing malignancies followed by a phase II study of patients with resected hepatic metastases of colon cancer to assess safety and feasibility of administering autologous DC loaded with CEA mRNA. The immunizations were well tolerated. Of the 24 evaluable patients in the dose escalation phase, there was 1 complete response (by tumor marker), 2 minor responses, 3 with stable disease, and 18 with progressive disease. In the phase II study, 9 of 13 patients have relapsed at a median of 122 days. Evidence of an immunologic response was demonstrated in biopsies of DC injection sites and peripheral blood of selected patients. We conclude that it is feasible and safe to administer mRNA-loaded DC to patients with advanced malignancies. PMID- 12901280 TI - A phase I study of gemcitabine and docetaxel for advanced stage solid tumors. AB - BACKGROUND: We conducted a phase I study to determine the maximum tolerated dose of docetaxel in combination with gemcitabine for patients with refractory solid tumors. METHODS: From January 1998 to November 1999, we treated 28 patients on a phase I protocol with gemcitabine given at a constant dose of 800 mg/m2 i.v. over 30 minutes on days 1, 8, and 15. Docetaxel was administered by a phase I schedule over 1 hour on day 1 of a 28-day cycle with a starting dose of 50 mg/m2 and increased by increments of 10 mg/m2 based on dose-limiting toxicity (DLT) that occurred in the first cycle. RESULTS: Neutropenia and thrombocytopenia were the dose-limiting toxicities. The maximum tolerated dose was 60 mg/m2. The most significant nonhematologic toxicities included fatigue, nausea, vomiting, mucositis, and hypersensitivity reactions. There was one partial response at 15 months in a patient with gastric cancer and six patients with stable disease for 4.0 to 15.0 months. CONCLUSIONS: The maximum tolerated dose of docetaxel with gemcitabine is 60 mg/m2. A Phase II study in selected primary sites is planned. PMID- 12901281 TI - Use of low-energy X-rays in the treatment of superficial nonmelanomatous skin cancers. AB - The Photon Radiosurgery System (PRS), developed by the Photoelectron Corporation in Lexington, Massachusetts, produces low-energy x-rays from the tip of a needle like probe at a high dose-rate. This portable device can be located directly over the target volume, delivering a uniform dose while sparing surrounding normal structures. To evaluate its radiotherapeutic profile, a prospective phase I Food and Drug Administration trial was performed utilizing this device in the management of nonmelanomatous skin cancers. Target doses ranged from 10 Gray in one fraction to 30 Gray in three weekly fractions. Thirty-eight lesions in 18 patients were treated. The overall completed response rate for all histologies at 12 months was 83%. No significant acute or late morbidity was encountered. Our results to date demonstrate that low-energy x-rays produced by the PRS were as effective in managing nonmelanomatous skin tumors as other radiation treatment modalities without the need for cumbersome shielding or radiation precautions. PMID- 12901282 TI - A comparison of three fatigue measures in veterans with cancer. AB - Fatigue is a highly prevalent and distressing symptom in cancer patients. The purpose of this study was to assess the validity of three fatigue measures [the Brief Fatigue Inventory (BFI), the Functional Assessment of Cancer Therapy Fatigue Subscale (FACT-F), and the lack of energy item from the Memorial Symptom Assessment Scale Short Form (MSAS-SF)] and compare these measures in relation to broader quality-of-life (QOL) constructs and clinical factors in veteran cancer patients. One-hundred-eighty cancer patients completed the BFI, FACT-F, FACT-G, MSAS-SF, and the Zung depression scale with concurrent Karnofsky performance status (KPS), laboratory tests, and demographic data. The Cronbach alpha coefficient was from 0.93 to 0.94 for BFI fatigue scales and 0.94 for FACT-F. There were significant correlations between BFI subscales, FACT-F, and lack of energy from MSAS-SF (p < 0.0001). All three fatigue measures showed significant correlation with MSAS-SF symptom subscales (p < 0.0001), FACT-G subscales (p < 0.0001), depression (p < 0.0001), KPS (p < 0.0001), inpatient status (P < 0.0001), insomnia (p < 0.05), hemoglobin (p < 0.05), and albumin levels (p < 0.01). Distress from lack of energy discriminated among levels from the BFI, FACT F, and FACT-G subscales and MSAS-SF subsclea by one-way of variance analysis. Patient responses to BFI, FACT-F, and the lack of energy item yielded similar information about broader QOL constructs and clinical factors. Single questions about lack of energy, or fatigue severity, may provide a simple and acceptable way to assess fatigue. PMID- 12901283 TI - Clinicians communicating with patients experiencing cancer pain. AB - PURPOSE: Provider-patient communication deficits are often implicated as barriers to adequate cancer pain relief. The purpose of this study was to describe verbal communication behaviors and interactions between providers and patients reporting cancer pain. METHODS: As part of a multisite clinical trial, we enrolled 17 oncology physician specialists and 84 patient participants who had reported cancer pain or treatment-related pain in the previous week associated with prostate or head and neck cancer. The study baseline clinic visits (N = 84) were audiotaped, transcribed, and entered into non-numerical unstructured data indexing searching and theorizing (NUD.IST) for content analysis. Each text unit in each transcript was coded as to conversation context: pain, additional symptom/side effects, tumor treatment, and/or personal remarks. Clinician questions were coded as either open-ended or closed-ended, clinician interruptions and subject changes were counted, and a measure of verbal dominance was calculated. RESULTS: The clinicians spent over half the conversations doing the talking. We calculated a close-ended to open-ended question ratio of 5.8:1. In 55% of the visits, the patient with cancer was interrupted by the clinician when the patient attempted to provide information or ask a question. Symptoms/side effects were addressed in practically all visits, whereas sensory pain was addressed in 90% of the visits. CONCLUSIONS: The clinicians were attentive to daily problems relevant to treatment side effects; however, the results also indicate a pattern of communication during the clinic visit that is typically clinician oriented. The nature of such communication may prevent the patient from sharing significant facts and experiences relevant to cancer pain and thus compromise the quality of pain management. PMID- 12901284 TI - Higher dose and dose-rate in smaller tumors result in improved tumor control. AB - Small tumors are more sensitive to radioimmunotherapy (RIT) than larger ones. A greater proportion of viable radiosensitive areas in small tumors, higher antibody uptake, and radiation dose may be responsible. Six groups of mice with small (median tumor size 0.06 cm3) or large LoVo xenografts (median tumor size 0.38 cm3) received either RIT using a 131I-labeled anti-CEA antibody A5B7, 5 fluorouracil (5-FU) modulated with folinic acid (FA), or no treatment. The % injected activity/gram, antibody distribution in viable and necrotic areas, and dose distribution were determined. High-power microscopy images of the original section were reconstructed to estimate the proportion of viable areas. Mice with small and large tumors grew significantly less rapidly when treated with RIT compared to the control group (p < 0.0004 and p < 0.003, respectively), while 5 FU was ineffective. Small tumors treated with RIT grew less than large tumors (p < 0.02). A higher amount of % injected activity/gram of tumor (median 26.6% vs. 8.1%, p = 0.0007) and a higher dose-rate were found in small tumors at 24 hours post injection (viable areas: 56.2 +/- 23.7 vs. 13.3 +/- 7 cGy/h, necrosis 19.2 +/- 16.3 vs. 4.9 +/- 4.7 cGy/h, p = 0.0007). It appears that as viable tumor masses grow the access to them decreases and this has a fourfold effect on dose delivered for RIT in this example. These data support the consideration of use of RIT for adjuvant treatment in colon cancer. PMID- 12901285 TI - The cytotoxicity of methyl protodioscin against human cancer cell lines in vitro. AB - Methyl protodioscin (NSC-698790) was a furostanol saponin isolated from the rhizome of Dioscorea collettii var. hypoglauca (Dioscoreaceae), a Chinese herbal remedy for the treatment of cervical carcinoma, carcinoma of the urinary bladder, and renal tumors for centuries. To systematically evaluate its potential anticancer activity, methyl protodioscin was tested cytotoxicity in vitro against human cancer cell lines by the NCI's (National Cancer Institute) anticancer drug screen. As a result, methyl protodioscin showed strong cytotoxicity against most cell lines from solid tumors with GI50 < or = 10.0 microM, especially selectively against one colon cancer line (HCT-15) and one breast cancer line (MDA-MB-435) with GI50 < 2.0 microM but moderate cytotoxicity was shown against leukemia cell lines with GI50 10-30 microM. The data are consistent with the fact that the rhizome of D. collettii var. hypoglauca has been employed for the treatment of solid tumors rather than leukemia in China for centuries. Based on an analysis using the COMPARE computer program with methyl protodioscin as a seed compound, no compounds in the NCI's anticancer drug screen database have cytotoxicity patterns similar to those of methyl protodioscin, indicating a potential novel mechanism of anticancer action. PMID- 12901286 TI - Lymphoblastoid alpha-interferon in the prevention of hepatocellular carcinoma (HCC) in high-risk HbsAg-positive resected cirrhotic HCC cases: a 14-year follow up. AB - Hepatocellular carcinoma (HCC) is a leading cause of death worldwide. It is also a common long-term complication of chronic hepatitis B or C infections, with cirrhosis as a risk factor for premalignant development. Natural lymphoblastoid alpha-interferon (alpha N1-IFN) has been widely used in treating chronic HBV and HCV carriers. To investigate its long-term beneficial effect in reducing the development of HCC, 20 hepatitis B surface antigen (HBsAg) positive resected cirrhotic HCC patients who have undergone chemotherapy followed by long-term treatment of 3MU alpha N1-IFN daily for 10 days every 3 months were analyzed in a 14 year follow-up. Results indicated that these patients survived and remained free of HCC. In contrast, 10 other patients receiving chemotherapy alone after resection eventually had HCC recur. Similar recurrence of HCC was seen in patients who had either reduced dosage of alpha N1-IFN or stopped interferon therapy or received 30MU dose at 6-month intervals. Our findings indicate the therapeutic potential and long-term safety of alpha N1-IFN in suppressing the development of HCC in high-risk patients. PMID- 12901287 TI - The androgen receptor: structure, mutations, and antiandrogens. AB - Androgens play a critical role not only in the physiological development of the prostate but also in the genesis of prostate cancer. The effects of androgen on the prostate gland and on the other tissues of the body are mediated by activation of the androgen receptor. The androgen receptor is a member of the superfamily of hormone receptors with a DNA-binding site, two zinc finger domains, and a hormone-binding site. Mutations in this receptor can be associated with loss of function or chronic endogeneous activation, depending upon the site of change. Androgens effect a conformal change in the structure of the androgen receptor associated with a change in protein phosphorylation. The androgen receptor can be activated by additional ligands affecting the hormone-binding site besides androgens. Activators and repressors of the androgen receptor modify this protein's function and are very delicately balanced such that disruptions of either function are associated with a disease state. Antiandrogens, which bind to the receptor and thus down-regulate the effects of endogeneous circulating androgens, remain the first line treatment for palliation of advanced prostate cancer. Mutations in the receptor are associated with a change in function of such compounds from antagonist to agonist in vitro. Newer evidence suggests there may be a role of intermittent androgen suppression rather than continuous suppression, perhaps by preventing overgrowth of hormone independent tumor cells. Future research focuses on the development of drugs directed at suppressing the androgen drive of the androgen sensitive clone of the tumor and making the nonsensitive subset more susceptible to cytotoxics. PMID- 12901288 TI - Hepatocellular carcinoma: are we making progress? AB - Hepatocellular carcinoma is a very prevalent malignancy worldwide, with increasing incidence in the United States. Despite many available treatment options, the prognosis remains poor. Surgical resection or liver transplantation still represents the only potentially curative treatments for HCC. Until more effective systemic therapies are available, different localized treatment approaches will continue to be applied in the management of this disease. Although systemic chemotherapy has been disappointing, increased understanding of the tumor biology in HCC coupled with new drug development may lead to newer agents with novel mechanisms of action that are more efficacious. The poor treatment outcome and dismal prognosis make prevention of HCC an important strategy in controlling this aggressive type of malignancy. Vaccine programs for HBV are ongoing. Efforts are underway to develop a vaccine for HCV. Interferon therapy appears to decrease the risk of developing HCC in patients with hepatitis, especially those with HCV. A number of other approaches for decreasing risk in these patients as well as in those with alcoholic-related cirrhosis are currently being evaluated. PMID- 12901289 TI - STI571 as a targeted therapy for CML. AB - Chronic myelogenous leukemia (CML) is a clonal hematopoietic stem cell disorder that progresses through distinct phases as the malignant clone progressively loses the capacity for terminal differentiation. It is characterized by the (9;22) translocation and resultant production of the Bcr-Abl tyrosine kinase. Bcr Abl functions as a constitutively activated tyrosine kinase, and this kinase activity is absolutely required for the transforming function of the Bcr-Abl protein. In preclinical studies, STI571 (Gleevec, imatinib mesylate), a Bcr-Abl tyrosine kinase inhibitor, specifically inhibited the proliferation of Bcr-Abl expressing cells in vitro and in vivo. STI571 has shown remarkable results in all phases of CML. Although responses are seen in all phases of the disease, durable responses are most common in earlier stage patients. Thus, STI571 has emerged as a paradigm for gene product targeted therapy, offering expanded treatment options for patients with CML. PMID- 12901291 TI - Cancers of the anal canal and anal margin. PMID- 12901290 TI - Chemotherapy-induced peripheral neuropathy. AB - While cancer remains an important public health concern, novel and enhanced treatment modalities have increased the length of survival of individuals diagnosed with the disease. The treatment of most cancers requires the use of chemotherapeutic agents to affect cure, maintain control of the disease, or provide palliation of symptoms. Although the use of chemotherapeutic agents can serve to prolong life, such agents are associated with significant side effects. Increasing clinical evidence suggests treatment of cancer with neurotoxic agents results in some degree of peripheral neuropathy. Specific drug categories implicated in the development of peripheral neuropathy are the plant alkaloids, interferons, antimitotics, taxanes, and platinum-based compounds. Drug-induced peripheral neuropathy is sensory, dose-related and cumulative and is usually delayed, appearing weeks after initiation of therapy. The number of individuals at risk for the development of chemotherapy-induced neuropathy is expected to increase proportionately with clinical protocols utilizing higher or more frequent dosing. As advanced cancer treatments and clinical trials can result in extending the lives of individuals affected by cancer, long-term functional deficits resulting from life-saving treatments must now be addressed. As such, peripheral neuropathy has emerged as an important consequence of cancer therapy. PMID- 12901292 TI - Predictive factors in prostate cancer: implications for decision making. PMID- 12901293 TI - Abandon all hope? The therapeutic misconception and informed consent. PMID- 12901294 TI - The role of the Deleted Colon Cancer (DCC) gene in colorectal and gastric cancer. PMID- 12901295 TI - A hope for the future. PMID- 12901297 TI - A half-century with solar neutrinos (Nobel Lecture). AB - The Sun derives its energy from fusion reactions in which hydrogen is transformed into helium. Every time four protons are turned into a helium nucleus, two neutrinos are produced. These neutrinos take only two seconds to reach the surface of the Sun and another eight minutes or so to reach the Earth. Thus, neutrinos tell us what happened in the center of the Sun eight minutes ago. The Sun produces one-third as many neutrinos as predicted by the standard solar model of particle physics. The author's pioneering work proved that nothing was wrong with the experiments or the theory; something was "wrong" with the neutrinos, in the sense that they behave in ways beyond the standard model. PMID- 12901296 TI - Predicting outcome in localized prostate cancer: how far have we come? PMID- 12901298 TI - Birth of neutrino astrophysics (Nobel Lecture). AB - The KamiokaNDE experiment for the observation of proton decay, an array of photomultipliers containing over 3000 tons of water, allowed the observation of charged particles travelling faster than the velocity of light in water. The subsequently developed Super-KamiokaNDE could be used to measure the amounts, the path, the energies, and the oscillation parameters of neutrinos, generated either by supernova explosions in the sun, or in the atmosphere. This work was awarded the 2002 Nobel Prize in Physics. PMID- 12901299 TI - The dawn of X-ray astronomy (Nobel Lecture). PMID- 12901300 TI - Metallobiliverdin radicals--DFT studies. AB - Several aspects of the molecular and electronic structure of biliverdin derivatives have been studied using density functional theory (DFT). The calculations have been performed for complexes of trianion (BvO2)3- and dianion [BvO(OH)]2-, derived from two tautomeric forms of biliverdin, BvO2H3 and [BvO(OH)]H2, with redox innocent metal ions: lithium(I), zinc(II), and gallium(III). One-electron-oxidized and reduced forms of each complex (cation and anion radicals) have been also considered. The molecular structures of all species investigated are characterized by a helical arrangement of tetrapyrrolic ligands with the metal ion lying in the plane formed by the two central pyrrole rings. The spin density distribution in four types of metallobiliverdin radicals- [(BvO2.)Mn+]n-2,[[BvO(OH).]Mn+]n-1 (cation radicals),[(BvO2.)Mn+]n 4,[[BvO(OH).]Mn+]n-3 (anion radicals)--has been investigated. In general, the absolute values of spin density on meso carbon atoms were larger than for the beta-carbon atoms. Sign alteration of spin density has been found for meso positions, and also for the beta-carbon atoms of at least two pyrrole rings. The calculated spin density maps accounted for the essential NMR spectroscopic features of iron biliverdin derivatives, including the considerable isotropic shifts detected for the meso resonances and shift alteration at the meso and beta positions. PMID- 12901301 TI - Cell adhesion onto highly curved surfaces: one-step immobilization of human erythrocyte membranes on silica beads. AB - This paper deals with single-step, orientation-selective immobilization of human erythrocyte membranes on bare silica beads with different topographies: 1) solid (nonporous) silica beads with a diameter of 3 microns and 2) porous silica beads with a diameter of 5 microns. Erythrocyte membranes were immobilized onto beads simply by incubation, without sonication or osmotic lysis. Membrane orientation before and after immobilization was identified with two immunofluorescence labels: 1) the extracellular part of glycophorin can be labeled with a first monoclonal antibody and a second polyclonal antibody with fluorescence dyes (outside label), while 2) the cytoplasmic domain of Band 3 can be recognized with a first monoclonal antibody and a second fluorescent polyclonal antibody (inside label). Adherent erythrocytes on the beads all ruptured, inverted the asymmetric orientation of the membrane, and selectively exposed their cytoplasmic domain. The surface topography did not influence the orientation or the amount of immobilized membrane. On the other hand, the fact that no adsorption or rupture of erythrocytes could be observed on planar quartz substrates suggests a significant influence of contact curvature on adhesion energy. PMID- 12901302 TI - Myristyl dimethylamine oxide surfactant solutions: model systems for rheological research. AB - Aqueous surfactant solutions of entangled, rod-shaped micelles are often characterized by monoexponential stress-relaxation processes. This special phenomenon leads to relatively simple theoretical descriptions, and viscoelastic surfactant solutions can, therefore, also be used as simple model systems for studying fundamental principles of flow. Herein, we present a detailed study of the nonlinear rheological properties of aqueous myristyl dimethylamine oxide surfactant solutions. In the regime of small deformations, shear stresses, or shear rates, the dynamic features of the viscoelastic solutions are characterized by the simple equations of a Maxwell material. At elevated values of shear stresses or deformations, however, this simple model fails and nonlinear features, such as normal stresses, stress overshoots, or shear-thinning properties occur. All these phenomena can be described by a Maxwell-type differential constitutive equation, which was first proposed by Giesekus. It turns out that the experimental results are in fairly good agreement with the theoretical predictions, if the anisotropy factor alpha is equal to 0.5. Besides transient data and nonlinear steady-state measurements, many semiempirical laws, such as the Cox-Merz rule, the Yamamoto relation, the Laun equation, and the Gleissble mirror relationships are approximately satisfied. The dynamic properties discussed in this paper are of general importance and they are equally observed in different materials such as polymer, dye, or protein solutions. PMID- 12901303 TI - Redox and conformational equilibria and dynamics of cytochrome c at high electric fields. AB - Cytochrome c (Cyt-c) adsorbed in the electrical double layer of the Ag electrode/electrolyte interface has been studied by stationary and time-resolved surface-enhanced resonance Raman spectroscopy to analyse the effect of strong electric fields on structure and reaction equilibria and dynamics of the protein. In the potential range between +0.1 and -0.55 V (versus saturated calomel electrode), the adsorbed Cyt-c forms a potential-dependent reversible equilibrium between the native state B1 and a conformational state B2. The redox potentials of the bis-histidine-coordinated six-coordinated low-spin and five-coordinated high-spin substates of B2 were determined to be -0.425 and -0.385 V, respectively, whereas the additional six-coordinated aquo-histidine-coordinated high-spin substate was found to be redox-inactive. The redox potential for the conformational state B1 was found to be the same as in solution in agreement with the structural identity of the adsorbed B1 and the native Cyt-c. For all three redox-active species, the formal heterogeneous electron transfer rate constants are small and of the same order of magnitude (3-13 s-1), which implies that the rate-limiting step is largely independent of the redox-site structure. These findings, as well as the slow and potential-dependent transitions between the various conformational (sub-)states, can be rationalized in terms of an electric field-induced increase of the activation energy for proton-transfer steps linked to protein structural reorganisation. Further increasing the electric field strength by shifting the electrode potential above +0.1 V leads to irreversible structural changes that are attributed to an unfolding of the polypeptide chain. PMID- 12901304 TI - Experimental and quantum-chemical studies on the thermochemical stabilities of mercury carbodiimide and mercury cyanamide. AB - Calorimetric dissolution measurements of the solid compounds mercury carbodiimide HgNCN(I) and mercury cyanamide HgNCN(II) in aqueous HCl that targeted at their thermochemical stabilities show the cyanamide species HgNCN(II) to be the more stable phase in terms of both enthalpy and Gibbs energy with an enthalpy difference of 2-3 kJ mol-1. While the stability ranking of HgNCN(I) and HgNCN(II) thus perfectly matches Pearson's HSAB concept, quantum-chemical stability predictions using common parametrizations of density functional theory appear to be fundamentally flawed. An analysis of the error is attempted on the basis of correlated wave functions for related molecules. PMID- 12901305 TI - Role of temperature in controlling performance of photorefractive organic glasses. AB - We present a detailed temperature-dependence study of dielectric, birefringent, conductive, and photorefractive (PR) properties of high-performance low-molecular weight organic glasses that contain 2-dicyanomethylene-3-cyano-2,5-dihydrofuran (DCDHF) derivatives. DCDHF organic glasses sensitized with C60 exhibit high two beam coupling gain coefficients in the red-wavelength region. However, in the best performing DCDHF glasses at room temperature the PR dynamics are limited by slow molecular reorientation in the electric field. While orientational and, therefore, PR speed can be significantly improved by increasing the temperature above the glass-transition temperature of the material, the steady-state performance may worsen. Comprehensive study of the temperature dependence of various processes, which contribute to the PR effect in DCDHF glasses, clarifies the limiting factors and allows for optimization of the overall PR performance. PMID- 12901306 TI - Triple FRET: a tool for studying long-range molecular interactions. PMID- 12901307 TI - Single- and multiple-molecule dynamics of the signaling from H-Ras to cRaf-1 visualized on the plasma membrane of living cells. PMID- 12901308 TI - G2 and DFT rigorous description of the inversion process of oxane and thiane used as simple ring systems to model sugar components. PMID- 12901309 TI - Second harmonic generation and fluorescence of CMONS dye nanocrystals grown in a sol-gel thin film. PMID- 12901310 TI - A simple aqueous-solution processing route to prepare quantum-confined CdS nanorods. PMID- 12901311 TI - Experimental and theoretical characterization of a triplet boron carbonyl compound: BBCO. PMID- 12901312 TI - The interplay of aryl-perfluoroaryl stacking interactions and interstack hydrogen bonding in controlling the structure of a molecular cocrystal. PMID- 12901313 TI - Pain barriers revisited: ten myths and misconceptions about pain management. PMID- 12901314 TI - The pressure is on for better blood pressure control. PMID- 12901315 TI - What I learned at legislative camp. PMID- 12901317 TI - Study reveals benchmarking flaws of many report cards, quality rankings. AB - Transferred patients have 38% longer ICU stays and 41% longer hospital stays. Many databases used for report cards are administrative, not clinical. Active awards are seen as more valid than passive ones. PMID- 12901316 TI - Assessment of knowledge and practice of high risk sexual behavior at a private Midwestern University. AB - In the United States, an estimated 15.3 million new cases of sexually transmitted infections (STIs) occur each year. Approximately two-thirds of people who acquire STIs in the U.S. are younger than 25 years of age. In addition, approximately 57% (3.1 million) of all pregnancies in the United States are classified as unintended. It is only logical that the consequences of high-risk sexual behavior (unintended pregnancy and contraction of an STI) would most affect those who exhibit high-risk sexual behavior. Thus, one would expect to find high-risk sexual behavior in the same populations that have high rates of STIs and unintended pregnancy. The purpose of this study was to survey freshman students living in the residence halls at a local college to assess their knowledge and the prevalence of high-risk sexual behavior, and give feedback to the participating institution about these results. The survey was designed to answer two questions: (1) What is the current level of high-risk sexual behavior at a small private Midwestern College? (2) What is the current level of knowledge about high-risk sexual behaviors of those surveyed? A total of 38 men and 131 women participated in the survey. Forty-seven percent of men and 41% of women respondents reported having sexual intercourse at least once in their lifetimes. Of those respondents who were sexually active, 67% of men and 54% of women reported that they "always use a condom" with sexual activity. Fifty-six percent of men and 51% of women that were sexually active reported that they were "not at all concerned" about contracting an STI. Twenty-two percent of men and 17% of women answered that they were "not at all concerned" about pregnancy. Three true/false questions about STIs were answered correctly over 95% of the time by those surveyed. Seventy-nine percent of respondents reported never having attended educational opportunities about STIs and safe sexual practices at the college. PMID- 12901318 TI - ADA, Kaiser using modeling in DM efforts. AB - Program predicted results of studies before data were even analyzed. Results of Archimedes program will be in public domain. Predictive modeling will be used to address most chronic diseases. PMID- 12901319 TI - Hotel shows health system keys to service excellence. AB - Ritz-Carlton partnership part of broader program pursuing service excellence. Nearly 600 system leaders will participate in day-long seminars. GE, Harvard serve as strategic partners for ongoing educational efforts. PMID- 12901321 TI - JCAHO OKs alternative safety goal approaches. PMID- 12901320 TI - Nurses and pharmacists partner for patient safety. AB - Professions face serious challenge of work force shortages. Leaders of five organizations come together in first step of ambitious journey. Revolutionary changes, not quick fixes, seen as solution. PMID- 12901322 TI - QI project cuts patients' chronic pain dramatically. PMID- 12901323 TI - Lupus anticoagulants: diagnosis and management. PMID- 12901324 TI - Reduced-intensity allogeneic stem cell transplantation. AB - The aim of reduced-intensity allogeneic stem cell transplantation is to exploit the graft-versus-tumor effect and reduce the toxicity associated with traditional ablative preparative regimens. Reduced-intensity regimens often use a purine analog with chemotherapeutic agents or low-dose total body irradiation. Postgrafting immunosuppression is variable and often consists of cyclosporine or tacrolimus plus methotrexate or mycophenolate mofetil. Regimen-related toxicity may be reduced using this approach, with a reduction in early mortality. Acute and chronic graft-versus-host disease continue to be problematic and are the primary causes of treatment-related mortality. The degree of myelosuppression is dependent on the regimen and the postgrafting immunosuppression. Myelosuppression is modest after 200 cGy of total body irradiation (with or without fludarabine), but it is more significant after regimens that use a purine analog with robust doses of chemotherapy. Stable mixed or full donor chimerism occurs with most regimens and graft rejection is more common in recipients of unrelated grafts. Reduced-intensity regimens are active in most hematologic malignancies and in some solid tumors, although the data in solid tumors are limited. Data suggest that rapidly growing malignancies require a more intensive regimen than indolent malignancies or those in remission. Reduced-intensity approaches have an appeal for nonmalignant diseases treated by allotransplantation. No prospective randomized trials have compared reduced-intensity approaches with conventional allotransplant preparation or postgrafting immunosuppression after reduced intensity allografting. However, the strategy holds great promise and offers the possibility of curative therapy for patients who are ineligible for an ablative transplant. PMID- 12901326 TI - Current treatment results of allogeneic bone marrow transplantation for acute myeloid and lymphoid leukemia. AB - Although acute myeloid and lymphoid leukemia initially respond to conventional chemotherapy, most patients relapse and succumb from their disease. For the past 30 years, efforts at intensifying induction or postremission conventional chemotherapy have met with limited success. Of all the therapies examined, allogeneic hematopoietic stem cell transplantation has achieved the best outcomes, with the first reports of cure 25 years ago. Most phase III studies have failed to demonstrate a clear advantage of allografting over chemotherapy because of significant risk of transplant-related mortality. However, there is reason for optimism based on the identification of high-risk groups and from improvements in the prevention and management of transplant-related complications. These accomplishments, along with a recent increase in the donor pool through the availability of unrelated donors and cryopreserved umbilical cord blood stem cells, continue to make allografting the most promising curative modality for acute leukemias. PMID- 12901325 TI - Graft-versus-host disease: emerging concepts in prevention and therapy. AB - Despite progress in allogeneic hematopoietic stem cell transplantation during the past three decades, graft-versus-host disease (GVHD) still contributes substantially to transplant-related morbidity and mortality. Although current strategies to prevent or treat this complication remain unsatisfactory, new insights into its pathophysiology have yielded novel targets for GVHD prophylaxis and therapy. Regulating GVHD without sacrificing beneficial graft-versus-tumor reactivity and immune competence continues to be an elusive goal. Many promising new interventions have undergone limited clinical evaluation, but controlled studies are needed to define their roles more clearly. Refinements in clinical grading systems and prognostic indices will facilitate the study of new strategies in GVHD prevention and treatment. PMID- 12901327 TI - Prevention and treatment of fungal infections in bone marrow transplantation. AB - There has not been as much success in the prevention and treatment of invasive fungal infections, particularly aspergillosis, compared to the prevention and treatment of cytomegalovirus infection and graft-versus-host disease in bone marrow transplant (BMT) recipients. Allogeneic BMT recipients who develop graft versus-host disease and remain immunosuppressed for long periods are at major risk for development of these infections. Prevention of environmental exposure, antifungal chemoprophylaxis, and attempts at early diagnosis are essential for the reduction of mortality from invasive fungal infections. Chest computerized axial tomography is extremely useful in diagnosing pulmonary aspergillosis. However, microbiologic or histologic identification of infection remains essential. Unfortunately, the response to therapy in BMT recipients remains suboptimal. With the development of the lipid formulations of amphotericin B, the newer azoles, and the echinocandins, safer and more efficacious options have become available. The optimal use of antifungal agents or their combinations remains to be determined. PMID- 12901328 TI - Autologous stem cell transplantation for non-Hodgkin's lymphoma. AB - Autologous stem cell transplantation (ASCT) is the only curative option for many patients with relapsed or refractory non-Hodgkin's lymphoma. ASCT achieves long term survival in up to 50% of patients with chemotherapy-sensitive relapsed diffuse large cell lymphoma (DLCL), and prospective randomized studies have documented the superiority of ASCT over salvage chemotherapy in patients with relapsed DLCL However, the role of ASCT as an upfront therapy for patients with high-risk DLCL remains unclear, and prospective randomized studies have yielded mixed results. In addition, ASCT may not be curative in follicle center or mantle cell lymphoma, although longer follow-up may identify a subset of patients with prolonged survival. Ongoing clinical trials are studying the use of monoclonal antibodies, such as rituximab and iodine 131I-tositumomab, in ASCT regimens to purge tumor cells in vivo and improve long-term outcome in follicle center and mantle cell lymphoma. PMID- 12901329 TI - Current status of gene therapy for hemophilia. AB - The hemophilias are an attractive model for gene therapy because their clinical manifestations are attributable to the lack of a single protein that circulates in minute amounts in the plasma. Sustained therapeutic expression of factors VIII and IX has been achieved in preclinical studies using a wide range of gene transfer technologies targeted at different tissues. This achievement has led to six different phase I/II clinical trials that resulted in limited efficacy but minimal toxicity. Recombinant adeno-associated viral vectors appear most promising for hemophilia gene therapy; however, this review summarizes all the major gene therapy approaches used and outlines the future challenges. PMID- 12901330 TI - Gene therapy for inherited immunodeficiencies. AB - Extensive research into gene therapy technologies has produced few clinically relevant results. Advances in the understanding of the genetics of inherited immunologic diseases, gene transfer methodologies, and stem cell manipulation have all contributed to successes in treating X-linked severe combined immunodeficiency and adenosine deaminase deficiency. This review examines the progress and the problems that have arisen, and discusses the improvement and future of gene therapy for primary immunodeficiencies. PMID- 12901331 TI - Gene therapy for fanconi anemia. AB - Fanconi anemia is a hereditary syndrome of bone marrow failure, congenital anomalies, and a predisposition to malignancy. Most patients die from bone marrow failure. Cells from patients display a heightened sensitivity to DNA cross linking agents with increased chromosomal breakage and increased cytotoxicity. Bone marrow from patients with Fanconi anemia have decreased numbers of hematopoietic progenitors when grown in culture. Transfer of the normal Fanconi anemia cDNA into cells from patients corrects the laboratory abnormalities, suggesting that gene transfer may prevent or reverse the bone marrow failure. Advances in gene transfer into human hematopoietic cells make this approach seem feasible. However, decreased numbers of stem cell targets may represent a significant obstacle. In addition, new insights on potential toxicities related to gene transfer have heightened a cautious approach. Fanconi anemia represents a prototype disorder for gene therapy and highlights the difficulties in adapting this technology to human disease. PMID- 12901332 TI - In vivo protection of hematopoietic cells from alkylator-mediated DNA damage. AB - Strategies that confer chemoresistance to hematopoietic stem and progenitor cells have two important future applications in the treatment of cancer and genetic diseases. Because dose-intensification of many cancer chemotherapy protocols is limited by severe hematopoietic toxicities, generation of primitive hematopoietic cells resistant to DNA damage mediated by chemotherapy may protect patients from life-threatening blood cytopenias. In addition, in the context of genetic diseases, overexpression of a chemoresistance gene in stem and progenitor cells may allow for the enrichment of small numbers of transduced cells that would not possess an in vivo selective advantage. In this report, I discuss studies that use the DNA repair protein O6-methylguanine DNA methyltransferase to protect hematopoietic cells from alkylator therapy. I focus on investigations evaluating the ability of O6-methylguanine-DNA methyltransferase mutant proteins to confer heightened resistance to alkylator-mediated DNA damage in vivo. PMID- 12901333 TI - Gene therapy for the hemoglobin disorders. AB - The hematologic disorders of beta-thalassemia and sickle cell disease are the most prevalent of human genetic diseases. Although palliative therapies and curative stem cell transplantation therapy have been developed for these disorders, treatment still remains suboptimal and many patients suffer significant morbidity and early mortality. Therefore, development of a gene therapy approach has been sought for many years. Major progress in the globin gene therapy field has been achieved by several laboratories. Using lentiviral vectors to obtain high-level expression of complex globin gene cassettes, therapeutic correction of several murine models of beta-thalassemia, and sickle cell disease was recently reported. This progress, coupled with developments in the ability to select and expand genetically modified stem cells in vivo, has advanced the possibility of gene therapy for the hemoglobin disorders in the near future. We review the developments in several areas that are critical for successful gene therapy of the hemoglobin disorders. PMID- 12901334 TI - Perspectives on gene therapy for lysosomal storage diseases that affect hematopoiesis. AB - Successful gene therapy for lysosomal storage diseases that involve the bone marrow and hematopoiesis is hindered by the hostile microenvironments created by these diseases and the irreversibility of the pathologic manifestations. Of the 40 lysosomal storage diseases, Gaucher disease type I, Niemann-Pick B, and mucopolysaccharidosis type I have hematopoietic involvement, are treatable by enzyme therapy, and respond to bone marrow transplantation. These storage diseases provide the basis for continued development of gene therapy. PMID- 12901336 TI - Electric field-induced effects on neuronal cell biology accompanying dielectrophoretic trapping. AB - Trapping neuronal cells may aid in the creation of the cultured neuron probe. The aim of the development of this probe is the creation of the interface between neuronal cells or tissue in a (human) body and electrodes that can be used to stimulate nerves in the body by an external electrical signal in a very selective way. In this way, functions that were (partially) lost due to nervous system injury or disease may be restored. First, a direct contact between cultured neurons and electrodes is created. This is realized using a microelectrode array (MEA) which can be fabricated using standard photolithographic and etching methods. Section 1 gives an overview of the human nervous system, methods for functional recovery focused on the cultured neuron probe, and the prerequisites for culturing neurons on a microelectrode array. An important aspect in the selective stimulation of neuronal cells is the positioning of cells or a small group of cells on top of each of the electrode sites of the MEA. One of the most efficient methods for trapping neuronal cells is to make use of di electrophoresis (DEP). Dielectrophoretic forces are created when (polarizable) cells are located in nonuniform electric fields. Depending on the electrical properties of the cells and the suspending medium, the DEP force directs the cells towards the regions of high field strength (positive dielectrophoresis; PDEP) or towards regions of minimal field intensities (negative dielectrophoresis; NDEP). Since neurons require a physiological medium with a sufficient concentration of Na+, the medium conductivity is rather high (~ 1.6 S/m). The result is that negative dielectrophoretic forces are created over the entire frequency range. With the use of a planar quadrupole electrode sturcture negative forces are directed so that in the center of this structure cell can be collected. The process of trapping cortical rat neurons is described in Sect. 2 theoretically and experimentally. Medium and cell properties are frequency dependent due to relaxation processes, which have a direct influence on the strength of the dielectrophorectic force. On the other hand, the nonideal material properties of the gold electrodes and glass substrate largely determine the electric field strength created inside the medium. Especially, the electrode medium interface results in a significant loss of the imput signal at lower frequencies (< 1 MHz), and thus a reduction of the electric field strength inside the medium. Furthermore, due to the high medium conductivity, the electric field causes Joule heating. Local temperature rises result in local gradients in fluid density, which induces fluid flow. The electrode-medium interface and induced fluid flow are theoretically investigated with the use of modeling techniques such as finite elements modeling. Experimental and theoretical results agreed with each other on the occurrence of the effects described in this section. For the creation of the cultured neuron probe, preservation of cell viability during the trapping process is a prerequisite. Cell viability of dielectrophoretically trapped neurons has to be investigated. The membrane potential induced by the external field plays a crucial role in preservation of cell viability. The membrane can effectively be represented by a capaticance in parallel woth a low conductance; with increasing frequency and /or decreasing field strength the induced membrane potential decreases. At high induced membrane potentials ths representation for the membrane is no longer valid. At this point membrane breakdown occurs and the normally insulating membrane becomes conductive and permeable. The creation of electropores has been proposed in literature to be the cause of this high permeability state. Pores may grow or many small pores may be created which eventually may lead to membrane rupture, and thus cell death. Membrane breakdown may be reversible, but a chemical imbalance created during the high permeability state may still exist after the resealing of the membrane. This may cause cell death after several hours or even days after field application. Section 3 gives a detailed description of membrane breakdown. Since many investigations on electroporation of lipid bilayers and cell membranes are based on uniform electric fields, a finite element model is used to investigate induced membrane potentials in the nonuniform field created by the quadropole electrode structure. Modeling results are presented in cmbination with the results of breakdown experiments using four frequencies in the range from 100 kHz to 1MHz. Radomly positioned neuronals cells were exposed to stepwise increasing electric field strengths. The field strength at which membrane rupture occurred gives an indication of the maximum induced membrane potential. Due to the nonuniformity of the electric field, cell collapse was expected to be position-dependent. However, at 100 kHz cells collapsed at a break down level of about 0.4 V, in contradistinction to findings at higher frequencies where more variation in breakdown levels were found. Model simulations were able to explain the experimental results. For examining whether the neuronal cells trapped by dielectrophoresis were still viable after the trapping process, the frequency range was divided into two ranges. First, a high frequency (14 MHz) and a rather low signal amplitude (3 Vpp) were used to trap cells. At this high frequency the field-induced membrane potential is small according to the theoretical model, and therefore no real damage is expected. The experimental analysis included the investigation of the growth of the neurons, number and length of the processes (dendrites and axons), and the number of outgrowing (~ viable) versus nonoutgrowing (~ nonviable) neural cells. The experimental results agreed with the expectation. The effect of the use of driving signals with lower frequencies and/or higher amplitudes on cell viability was investigated using a staining method as described in the second part of Sect. 4. Survival chances are not directly linked to the estimated maximum induced membrane potential. The frequency of the dield plays an important role, decreasing frequency lowering the chance of survival. A lower frequency limit of 100 kHz is preferable at field strengths less than 80 k V/m, while with increasing field strength this limit shifts towards higher frequencies. The theoretical and experimental results presented in this review form the inception of the development of new electrode structures for trapping neuronal cells on top of each of the electrodes of the MEA. New ways to investigate cell properties and the phenomenon of electroporation using electrokinetic methods were developed that can be exploited in future research linking cell biology to technology. PMID- 12901335 TI - The sensory circumventricular organs of the mammalian brain. AB - The brain's three sensory circumventricular organs, the subfornical organ, organum vasculosum of the lamina terminalis and the area postrema lack a blood brain barrier and are the only regions in the brain in which neurons are exposed to the chemical environment of the systemic circulation. Therefore they are ideally placed to monitor the changes in osmotic, ionic and hormonal composition of the blood. This book describes their. General structure and relationship to the cerebral ventricles Regional subdivisions Vasculature and barrier properties Neurons, glia and ependymal cells Receptors, neurotransmitters, neuropeptides and enzymes Neuroanatomical connections Functions. PMID- 12901337 TI - Autologous stem cell transplantation in multiple myeloma: none, one, or two? PMID- 12901338 TI - The myelodysplastic syndromes: classification and prognosis. AB - Myelodysplastic syndrome is a neoplastic clonal stem cell disorder characterized clinically by bone marrow failure and a tendency to progress to acute myelogenous leukemia. Dysplasia is the pathologic hallmark. The French-American-British classification served as the gold standard for more than two decades. Under the auspice of the World Health Organization, more than 100 hematopathologists in a 3 year cumulative effort issued the new World Health Organization classification, which recognizes multilineage dysplasia. Refractory anemia with excess blasts is divided into two groups. Chronic myelomonocytic leukemia is reclassified under a separate category. Refractory anemia with excess blasts in the transformation group was omitted. Finally, 5q-syndrome is a new subgroup. In addition to the pathologic classification, various prognostic predictors were formatted into scoring systems. Bone marrow blast percentage, cytopenias, and cytogenetics are the backbone for those prognostic models. The International Prognostic Scoring System is a product of pooled data from previous scoring systems and a useful tool to predict survival and acute myelogenous leukemia evolution. This paper discusses the classification and prognosis of myelodysplastic syndromes and their evolution. PMID- 12901339 TI - Apoptosis in the myelodysplastic syndromes. AB - Myelodysplastic syndromes (MDS) are neoplastic dyscrasias characterized by peripheral cytopenia, despite a normocellular or hypercellular bone marrow. In the past decade, it has become apparent that this ineffective hemopoiesis is largely caused by excessive apoptosis of myeloid precursors. There is no evidence for gain-of-function mutations within the apoptotic machinery in MDS. It appears that the apoptosis is a reactive phenomenon fueled by cytokines. The provoking stimulus for the proapoptotic intramedullary milieu in MDS is unknown. The evolution of MDS from early relatively chronic to aggressive and frankly leukemic phenotype is accompanied by a suppression of apoptosis. This metamorphosis correlates with changes in intracellular levels of Bcl-2-family proteins, but the genetic basis for this shift has not been elucidated clearly. Expression profiling and proteomic technologies may offer the best means to unravel this process. PMID- 12901340 TI - Emerging therapeutic strategies for myelodysplastic syndrome. AB - Increasing understanding of the pathophysiology of myelodysplastic syndromes (MDS) and a greater appreciation of the large impact that marrow failure has on the symptomatology and outcome of MDS have led to new treatment approaches. We describe recent developments of methods to manage marrow failure, suppress the MDS clone, and widen the applicability of curative treatment with allogeneic stem cell transplants. PMID- 12901341 TI - CMML: a biologically distinct myeloproliferative disease. AB - Chronic myelomonocytic leukemia (CMML) is a myeloproliferative disorder with unique characteristics. Dysplasia is usually present in the bone marrow, thus CMML has usually been classified as a myelodysplastic syndrome. The recent World Health Organization classification of myeloid malignancies proposes to classify CMML into a new category of myelodysplastic syndromes and myeloproliferative disorders. A new prognostic score has also been developed exclusively for patients with CMML that recognizes four groups with distinct prognoses. Significant biologic findings in the recent months include the recognition of the importance of angiogenesis in CMML with a possible autocrine role for vascular endothelial growth factor, and the further understanding of the role of tyrosine kinase fusion genes and activation in some patients with CMML Therapeutic discoveries have been hampered by the paucity of studies looking specifically at CMML Among agents with potential significant activity are imatinib mesylate (for patients with platelet-derived growth factor beta receptor-associated fusion genes), hypomethylating agents, antiangiogenic agents, farnesyltransferase inhibitors, and topoisomerase I inhibitors. Future studies should consider CMML as a separate entity to promote a better understanding and identify more effective therapy for patients with this disease. PMID- 12901342 TI - Hemopoietic cell transplantation for myelodysplastic syndromes. AB - Myelodysplastic syndromes are clonal stem cell disorders, and allogeneic hemopoietic cell transplantation (HCT) is the only therapy with curative potential. Among patients with less-advanced myelodysplastic syndromes, 3-year survivals of 65% to 75% are achievable with human leukocyte antigen (HLA) identical related and HLA-matched unrelated donors. The probability of relapse is less than 5%. Among patients with advanced disease (> or = 5% marrow blasts), 35% to 45% and 25% to 30%, respectively, are surviving in remission after transplantation from related or unrelated donors. The incidence of post transplant relapse is 10% to 35%. Criteria of the International Prognostic Scoring System, originally developed for nontransplanted patients, also predict relapse and survival after HCT. Refined conditioning regimens have permitted successful HCT even in patients in their seventh decade of life. Results with a regimen using a combination of busulfan (targeted to predetermined plasma levels) and cyclophosphamide are particularly encouraging. Improved survival with transplants from unrelated volunteer donors reflects selection of donors on the basis of high-resolution (allele-level) HLA typing. Nevertheless, transplant related morbidity and mortality, including graft-versus-host disease, remain challenges that need to be addressed with innovative approaches. Some patients who achieve a chemotherapy-induced complete remission may also benefit from autologous HCT. PMID- 12901343 TI - The chronic myeloproliferative disorders: an historical perspective. AB - To comprehend the present and chart an informed course for the future, it is essential to be aware of the past. Fifty years ago, William Dameshek introduced the conceptual construct of myeloproliferative disorders, a watershed event in the evolution of thought regarding that peculiar cluster of similar marrow conditions. The four major disorders that comprise the myeloproliferative group, polycythemia vera, chronic myeloid leukemia, myelofibrosis with myeloid metaplasia, and essential thrombocythemia, have interesting and instructive histories. This narrative review explores some of the most noteworthy episodes in those stories, from first descriptions to current understanding. PMID- 12901344 TI - PRV-1 mRNA expression and other molecular markers in polycythemia rubra vera. AB - During the past 4 years, the first molecular markers for polycythemia vera (PV) have been described. These markers include decreased expression of the thrombopoietin receptor c-Mpl, increased expression of the surface receptor polycythemia rubra vera-I, and insulin-like growth factor-I hypersensitivity. Characterization of these abnormalities has allowed the development of the first molecular diagnostic tools for PV. However, despite these advances, the molecular mechanisms leading to the development of PV remain poorly understood. This review summarizes and evaluates recent advances in our understanding of molecular aberrations in PV. PMID- 12901345 TI - A contemporary approach to the diagnosis and management of polycythemia vera. AB - The natural history of polycythemia vera (PV) includes an increased lifetime risk of thrombohemorrhagic complications and disease transformation into myelofibrosis with myeloid metaplasia and acute myeloid leukemia. The latter is the primary reason for the shortening of survival that becomes significant after the first decade of disease. Historic nonrandomized studies have suggested that aggressive phlebotomy improves survival in PV. However, prospective randomized studies have failed to demonstrate a better treatment than phlebotomy alone, in terms of survival. However, the addition of cytoreductive therapy to phlebotomy in high risk patients with PV may reduce the risk of recurrent thrombosis. Other disease features of PV include aquagenic pruritus and microvascular disturbances such as erythromelalgia. This review outlines a practical approach to diagnosis, in addition to treatment of life-threatening and non-life-threatening complications of PV. PMID- 12901346 TI - Pathogenic markers in essential thrombocythemia. AB - Although long recognized as a clinical entity, the diagnosis of essential thrombocythemia (ET) still relies on the exclusion of reactive conditions and other myeloproliferative disorders. Recent studies have attempted to identify new markers for this disorder and suggest that part of the problem may relate to its heterogeneity. Cytogenetic abnormalities are rare, but analysis using X chromosome inactivation patterns indicates that nearly 50% of evaluable patients have polyclonal myelopoiesis and this is associated with a lower risk of thrombosis. Reduced and heterogeneous staining of c-Mpl in bone marrow biopsies may help to distinguish ET and a reactive thrombocytosis. Increased expression of PRV-1 (polycythemia ruba vera) in granulocytes may assist in discriminating between polycythemia vera and at least some cases of ET. The contribution of these markers to disease pathogenesis is unknown and prospective studies are needed to evaluate their usefulness for predicting clinical outcome and directing patient therapy. PMID- 12901347 TI - Treatment indications and choice of a platelet-lowering agent in essential thrombocythemia. AB - The therapeutic strategy in patients with essential thrombocythemia (ET) is a difficult balance between the prevention of bleeding and thrombotic complications and the risks of drug side effects and toxicity. Major bleeding is rare and seems to be related to higher platelet counts. Therefore, a platelet count greater than 1500 x 10(9)/L is generally regarded as an indication for cytoreduction. Thrombotic complications include microvascular occlusive symptoms, which are usually reversible with low-dose aspirin, and large vessel thrombosis. The risk of major thrombosis is higher in patients with ET who are older than 60 years and who have had a previous occlusive event. In this high-risk group, the nonalkylating agent hydroxyurea significantly reduces the rate of vascular complications and has emerged as the treatment of choice. However, the long-term risk:benefit ratio of hydroxyurea remains disputed because its leukemogenic potential has not been ruled out. This holds also for other myelosuppressive agents such as busulfan and pipobroman. Other drugs of particular interest for young patients include recombinant interferon-alpha and anagrelide. Both of these drugs are effective in lowering platelet counts, but their efficacy in reducing clinical complications remains to be demonstrated. Furthermore, interferon and anagrelide have frequent and clinically important side effects. Thus, further clinical studies are required to establish their role in the management of patients with ET. PMID- 12901348 TI - Critical review of pathogenetic mechanisms in myelofibrosis with myeloid metaplasia. AB - Myelofibrosis with myeloid metaplasia is an uncommon chronic myeloproliferative disorder characterized by extramedullary hematopoiesis associated with varying degrees of bone marrow fibrosis (hematopoiesis is clonal and fibrosis is a polyclonal reactive process). The primary defect in a pluripotent stem cell is still unknown. However, advances have been made during the past few years in the knowledge of the pathogenetic mechanisms in this disorder. This review focuses on current data on the cellular and molecular mechanisms possibly participating in the development of the disease. PMID- 12901349 TI - Myelofibrosis with myeloid metaplasia: therapeutic options in 2003. AB - Myelofibrosis with myeloid metaplasia (MMM) is a progressive, clonal, and lethal myeloproliferative disorder with a variable prognosis. The only potentially curative therapy for MMM is allogeneic stem cell transplantation, but this therapy is too toxic for older patients and inappropriate for young patients with good prognostic features. A nonmyeloablative or autologous stem cell transplant may broaden the applicability of high-dose therapy for MMM, but the therapy for MMM is palliative in intent and aimed at symptomatic relief. There is no approved therapy for MMM; however, certain agents have been useful for targeted palliation. Myelosuppressive drug therapy, targeted radiation, or splenectomy has decreased symptomatic myeloproliferation (i.e., splenomegaly). MMM-associated cytopenias may be improved by the selective use of growth factors, androgens, and thalidomide. Debilitating constitutional symptoms improve with the pharmacologic blockage of tumor necrosis factor-alpha. Further molecular understanding of MMM is necessary for more effective targeted therapies. PMID- 12901350 TI - [Infectious arthtropathy--destiny or indication for endoprosthetic joint replacement]. PMID- 12901351 TI - [Shoe modification or inserts can effect relief of the knee joint]. PMID- 12901352 TI - [Secondary arthropathy after arthrodesis of the lower ankle joint]. PMID- 12901353 TI - Finance issue brief: women's health: osteoporosis coverage requirements: year end report-2002. PMID- 12901354 TI - Finance issue brief: women's health: inpatient mastectomy length of stay & breast reconstructive surgery coverage requirements: year end report-2002. PMID- 12901355 TI - Finance issue brief: women's health: breast & cervical cancer screening coverage requirements: year end report-2002. PMID- 12901356 TI - Inhibition of tumor angiogenesis by angiostatin: from recombinant protein to gene therapy. AB - Tumor growth, local invasion, and metastatic dissemination are dependent on the formation of new microvessels. The process of angiogenesis is regulated by a balance between pro-angiogenic and anti-angiogenic factors, and the shift to an angiogenic phenotype (the "angiogenic switch") is a key event in tumor progression. The use of anti-angiogenic agents to restore this balance represents a promising approach to cancer treatment. Known physiological inhibitors include trombospondin, several interleukins, and the proteolytic break-down products of several proteins. Angiostatin, an internal fragment of plasminogen, is one of the more potent of this latter class of angiogenesis inhibitors. Like endostatin, another anti-angiogenic peptide derived from collagen XVIII, angiostatin can induce tumor vasculature regression, leading to a complete cessation of tumor growth. Inhibitors of angiogenesis target normal endothelial cells, therefore the development of resistance to these drugs is unlikely. The efficacy of angiostatin has been demonstrated in animal models for many different types of solid tumors. Anti-angiogenic cancer therapy with angiostatin requires prolonged administration of the peptide. The production of the functional polypeptides is expensive and technical problems related to physical properties and purity are frequently encountered. Gene transfer represents an alternative method to deliver angiostatin. Gene therapy has the potential to produce the therapeutic agent in high concentrations in a local area for a sustained period, thereby avoiding the problems encountered with long-term administration of recombinant proteins, monoclonal antibodies, or anti-angiogenic drugs. In this review we compare the different gene therapy strategies that have been applied to angiostatin, with special regard to their ability to provide sufficient angiostatin at the target site. PMID- 12901357 TI - The response of endothelial cells to fluid shear stress using a co-culture model of the arterial wall. AB - An endothelial cell (EC) smooth muscle cell (SMC) co-culture model of the arterial wall was used to study the effect of fluid shear stress on EC behavior. This model, in addition to being a more realistic tissue analogue, is a valuable research tool for studying the effects of mechanical stimulation upon the behavior of both SMCs and ECs. In the present study, a 10% cyclic strain was used to alter the characteristics of an SMC-seeded collagen gel. This form of strain preconditioning resulted in a rearrangement of the vessel wall that yielded circumferentially oriented cells and collagen fibrils. The preconditioned collagen gel was subsequently seeded with ECs and exposed to fluid-induced shear stress (10 dynes/cm2) for 48 hr. In the absence of flow, ECs seeded on slab constructs were oriented with the underlying collagen fibrils. Sheared constructs exhibited ECs oriented in the flow direction. Shear stress also affected EC proliferation, reducing the total number of dividing ECs by as much as 48 percent compared to unsheared constructs. The shear-induced reduction in proliferation was further enhanced when constructs were first strain-preconditioned (64% reduction). Moreover, conditioned media from shear stress experiments inhibited proliferation of ECs seeded on tissue culture plastic. These results suggest that EC response to fluid shear stress in a collagen co-culture model is influenced by the underlying substrate, and one that in this study is modified by strain preconditioning. PMID- 12901358 TI - Augmented binding and activation of latent transforming growth factor-beta by a tryptic fragment of latency associated peptide. AB - Transforming growth factor-beta (TGF-beta) is secreted in a latent form; thus, activation is critical for the control of TGF-beta action. Latent TGF-beta exists in a complex in which mature TGF-beta is noncovalently linked to latency associated peptide (LAP) and latent TGF-beta binding protein (LTBP) complex. We have shown that latent TGF-beta is efficiently activated in heterotypic cultures of endothelial cells (ECs) and smooth muscle cells (SMCs). Under those conditions, LAP plays an important role in targeting latent TGF-beta to the surface of SMCs, and plasmin and calpain target it to the surface of ECs for activation. Here, we demonstrate in a homotypic culture system that fragments of LAP increase the binding of latent TGF-beta to ECs, resulting in its activation by cell-associated proteolysis. LAP fragments appear to bind to the cell surface and augment the binding of latent TGF-beta, independent of transglutaminase. These results suggest a unique mechanism for the activation of latent TGF-beta by proteolytic fragments of LAP. The mechanism may arise from degradation by elevated levels of proteases under certain conditions. PMID- 12901359 TI - Metabolism of dynorphins by peptidases of pulmonary artery endothelial cells. AB - Degradation of several dynorphins by peptidases expressed in cultured porcine pulmonary artery endothelial cells was studied by incubation of the peptide in cell suspensions followed by electrospray ionization and tandem mass spectrometric analyses. Under the in vitro conditions applied, only the metabolism of dynorphin A1-8 occurred in a significant extent. Studies involving specific peptidase inhibitors indicated that mainly bestatin-sensitive aminopeptidases, thiorphan-sensitive endopeptidases, and cFPAAF-pAB-sensitive endopeptidases expressed by the endothelial cells were involved in the process that converted dynorphin A1-8 to dynorphin A2-8, dynorphin A1-6, and leucine enkephalin (dynorphin A1-5), respectively. These peptidases may form a metabolic barrier for the cellular penetration of intact dynorphin A1-8 and/or control effects of the circulating peptide on endothelial opioid receptors of the cells. PMID- 12901360 TI - Intracellular Ca2+ homeostatic regulation and 4-hydroxynonenal-induced aortic endothelial dysfunction. AB - The aldehydic lipid peroxidation product 4-hydroxynonenal (HNE) is known to compromise erythrocyte passive Ca2+ permeability and to irreversibly inhibit the plasma membrane (Ca2+ + Mg2+)-ATPase and Ca2+-transport. To measure the effects of HNE on passive and active Ca2+ transport in endothelial cells, we first characterized 45Ca2+ uptake and efflux in cultured porcine aortic endothelial cells (PAEC). PAEC exchanged 45Ca2+ to a cumulative near-isotopic equilibrium of about 4.5 pmole 45Ca2+/10(6) cells in 120 min at 37 degrees C. This Ca2+ pool was diminished by thapsigargin, cyclopiazonic acid, oligomycin B, and sodium azide. In contrast, ouabain enhanced Ca2+ uptake capacity from 5.17 to 5.77 pmole/10(6) cells. Accumulated 45Ca2+ was extruded at rate of 8.7 fmole 45Ca2+/10(6) cells/min or shunted rapidly by the ionophore A23187. HNE increased total 45Ca2+ accumulation in a time- and concentration-dependent manner by as much as 562% with an EC50 of 64.0 wM. Concomitant morphological analysis of PAEC revealed vacuolization, nuclear swelling, cell shrinking, and cell detachment. Initial structural changes, such as vacuolization, began well before any changes in Ca2+ accumulation were observed. These functional and morphological changes indicate that HNE significantly increases intracellular Ca2+ accumulation in vascular endothelium, which may explain the cytotoxic effects associated with HNE exposure and provide further evidence that atherogenic effects of HNE may, in part, be caused by disturbances in Ca2+ homeostasis. PMID- 12901361 TI - The 15th Annual Meeting of the Winter Conference on Neural Plasticity, Guadeloupe, French Antilles, February 23-28, 2003. PMID- 12901363 TI - Medical image. Disabling acrocyanosis. PMID- 12901362 TI - Re: Gale G, Nussbaum D, Rothbart P, Hann B, Leung V, Kanetz G. A randomized treatment study to compare the efficacy of repeated nerve blocks with cognitive therapy for control of chronic head and neck pain. Pain Res Manage 2002;7:185-9; and Merskey H, Thompson EN. Nerve blocks and cognitive failure: A beneficial failure. Pain Res Manage 2002;7:175-6. PMID- 12901364 TI - 6-thioguanine nucleotides and thiopurine methyltransferase activity: important factors determining response to treatment and incidence of adverse effects from azathioprine and 6-MP. PMID- 12901365 TI - Proceedings of the 28th Symposium on Nucleic Acids Chemistry. Yokohama, Japan, November 7-9, 2001. PMID- 12901367 TI - The view from Les Treilles on the origins, evolution and diversity of viruses. PMID- 12901366 TI - The great virus comeback-- from an evolutionary perspective. PMID- 12901368 TI - [The impact of a planning intervention to improve the consistency of drug prescription issuing rules in a health facility]. PMID- 12901369 TI - [Therapy with epoetin beta: safe and effective in prevention and treatment of tumor-induced anemia]. PMID- 12901371 TI - NASA Astrobiology Institute general meeting. February 10-12, 2003, Tempe, Arizona, USA. Abstracts. PMID- 12901370 TI - About veterinary education in Iraq. AB - The cons and pros of veterinary education in Iraq are described. Started as a small institution, with few students and with foreign staffs, then expanded to enroll more than hundred students each year, with all Iraqi staff. The graduates of the Veterinary College played an important role in monitoring animal health, supervising research projects involving animal welfare, some served as educators of various veterinary science specializations, others worked as private practitioners or recruited in the army. Veterinary education was very vital, as other sciences for progress of the country. PMID- 12901372 TI - Museum of history of veterinary medicine, Kosice - Slovakia. PMID- 12901374 TI - [Domestic animals in heraldry, volume1]. AB - The author presents in the first part of her paper the historical background for the heraldic. In the coming seven papers she will discuss the use and position of different domestic animals in the heraldic. PMID- 12901373 TI - Parallel evolution by gene duplication in the genomes of two unicellular fungi. AB - Phylogenetic analysis of conserved gene families in fission yeast Schizosaccharomyces pombe and brewer's yeast Saccharomyces cerevisiae showed that gene duplications have occurred independently in the same families in each of these two lineages to a far greater extent than expected by chance. These species represent distinct lineages of the phylum Ascomycota that independently evolved a "yeast" life cycle with a unicellular thallus that reproduces by budding, and many of the genes that have duplicated independently in the two lineages are known to be involved in crucial aspects of this life cycle. Parallel gene duplication thus appears to have played a role in the independent origin of similar adaptations in the two species. The results indicate that using phylogenetic analysis to test for parallel gene duplication in different species may help in identifying genes responsible for similar but independently evolved adaptations PMID- 12901375 TI - [The influence of the moon on bleeding]. AB - The theory of the influence of the stars and in particular of the moon to bleeding was most important in the 17th century; However, a survival of this belief was noted during springtime bleeding or treatment of mare sterility, seldom on other circumstances, in some traditional regions of France from 1930 to 1960. The results presented in the paper are obtained 1990-91 by enquiring of approx. 3.500 French veterinarians graduated 1911-1957. PMID- 12901376 TI - [Adolpho Lutz: a biographical sketch]. AB - This article portrays the family origins and life story of Adolpho Lutz (1855 1940) up to his transfer to the Instituto Oswaldo Cruz in 1908. His life history is used as a motif for an analysis of the institution of pasteurian and tropical medicine in Brazil. His university and postgraduate study in German-speaking Europe are examined, as are his activities as a clinician and researcher on subjects related to helminthology, parasitology, veterinary medicine and bacteriology in the interior of Sao Paulo state; his stay at the Molokai leprosarium in Hawaii; and the medical controversies in which he participated as head of the Bacteriological Institute of Sao Paulo, especially those on cholera, dysenteries, typhoid fever, malaria and yellow fever. PMID- 12901377 TI - [On the abominable profession of being a vampire: Emilio Goeldi and Mosquitos in Para (1905)]. AB - In the fields of public health and entomology, the turn of the 20th century was marked by an emphasis on insects as transmitters of disease. Physicians, bacteriologists and zoologists perseverently tried to understand the etiology of diseases like malaria and yellow fever and make an inventory of the species of insects associated with disease, studying their scientific and biological classification and establishing procedures to control epidemics. The Swiss zoologist Emilio Goeldi (1859-1917), who was then director of the Museum of Natural History and Ethnography of Para, in Belem, was an active participant in this scientific network and wrote a number of papers on mosquitos in the Amazon. His ideas and contributions to zoology and medical entomology are the focus of this paper. PMID- 12901379 TI - [Epistemological frameworks of tropical medicine]. AB - In order to understand the novelty, in theoretical and epistemelogical terms, of the emergence of tropical medicine, we analyse two models of research represented by the Pasteur de Ultramar Institute and the London School of Tropical Medicine. We seek to understand which arguments and concepts come into play in their discourse; in what way they construct their protocols for research; how these protocols, concepts and arguments are similar and different; indeed, whether it is possible to speak of something new, epistemologically speaking, or of the incommensurability of tropical medicine with the medicine inherited from Pasteur and the medical tradition concerned with disease in hot climates. PMID- 12901380 TI - [Salvador, world city: from Germany to Bahia]. AB - This paper discusses the German presence in 19th century Bahia. There was an intense flow of foreigners to the city of Salvador and other parts of what was then a province; these people were involved in trade, farming, industry and services. Among the Germans who crossed the Atlantic to establish themselves in Bahia, medical doctors are a particularly interesting object of study, along with their motives for going to America and how they worked in the field of scientific knowledge at the time. The paper highlights studies by Otto Wucherer and his activities as a clinician, researcher and agent of the intersection between the German scientific and the Bahian communities. PMID- 12901381 TI - Nurses in the maternity services. PMID- 12901378 TI - Alteration of hepatitis A virus (HAV) particles by a soluble form of HAV cellular receptor 1 containing the immunoglobin-and mucin-like regions. AB - Hepatitis A virus (HAV) infects African green monkey kidney cells via HAV cellular receptor 1 (havcr-1). The ectodomain of havcr-1 contains an N-terminal cysteine-rich immunoglobin-like region (D1), followed by a mucin-like region that extends D1 well above the cell surface. D1 is required for binding of HAV, and a soluble construct containing D1 fused to the hinge and Fc portions of human immunoglobin G1 (IgG1), D1-Fc, bound and neutralized HAV inefficiently. However, D1-Fc did not alter the virions. To determine whether additional regions of havcr 1 are required to trigger uncoating of HAV, we constructed D1muc-Fc containing D1 and two-thirds of the mucin-like region fused to the Fc and hinge portions of human IgG1. D1muc-Fc neutralized 10 times more HAV than did D1-Fc. Sedimentation analysis in sucrose gradients showed that treatment of HAV with 20 to 200 nM D1muc-Fc disrupted the majority of the virions, whereas treatment with 2 nM D1muc Fc had no effect on the sedimentation of the particles. Treatment of HAV with 100 nM D1muc-Fc resulted in low-level accumulation of 100- to 125S particles. Negative-stain electron microscopy analysis revealed that the 100- to 125S particles had the characteristics of disrupted virions, such as internal staining and diffuse edges. Quantitative PCR analysis showed that the 100- to 125S particles contained viral RNA. These results indicate that D1 and the mucin-like region of havcr-1 are required to induce conformational changes leading to HAV uncoating. PMID- 12901382 TI - Iterative pass optimization of sequence data. AB - The problem of determining the minimum-cost hypothetical ancestral sequences for a given cladogram is known to be NP-complete. This "tree alignment" problem has motivated the considerable effort placed in multiple sequence alignment procedures. Wheeler in 1996 proposed a heuristic method, direct optimization, to calculate cladogram costs without the intervention of multiple sequence alignment. This method, though more efficient in time and more effective in cladogram length than many alignment-based procedures, greedily optimizes nodes based on descendent information only. In their proposal of an exact multiple alignment solution, Sankoff et al. in 1976 described a heuristic procedure--the iterative improvement method--to create alignments at internal nodes by solving a series of median problems. The combination of a three-sequence direct optimization with iterative improvement and a branch-length-based cladogram cost procedure, provides an algorithm that frequently results in superior (i.e., lower) cladogram costs. This iterative pass optimization is both computation and memory intensive, but economies can be made to reduce this burden. An example in arthropod systematics is discussed. PMID- 12901383 TI - Implied alignment: a synapomorphy-based multiple-sequence alignment method and its use in cladogram search. AB - A method to align sequence data based on parsimonious synapomorphy schemes generated by direct optimization (DO; earlier termed optimization alignment) is proposed. DO directly diagnoses sequence data on cladograms without an intervening multiple-alignment step, thereby creating topology-specific, dynamic homology statements. Hence, no multiple-alignment is required to generate cladograms. Unlike general and globally optimal multiple-alignment procedures, the method described here, implied alignment (IA), takes these dynamic homologies and traces them back through a single cladogram, linking the unaligned sequence positions in the terminal taxa via DO transformation series. These "lines of correspondence" link ancestor-descendent states and, when displayed as linearly arrayed columns without hypothetical ancestors, are largely indistinguishable from standard multiple alignment. Since this method is based on synapomorphy, the treatment of certain classes of insertion-deletion (indel) events may be different from that of other alignment procedures. As with all alignment methods, results are dependent on parameter assumptions such as indel cost and transversion:transition ratios. Such an IA could be used as a basis for phylogenetic search, but this would be questionable since the homologies derived from the implied alignment depend on its natal cladogram and any variance, between DO and IA + Search, due to heuristic approach. The utility of this procedure in heuristic cladogram searches using DO and the improvement of heuristic cladogram cost calculations are discussed. PMID- 12901384 TI - Endocrine-related resources from the National Institutes of Health. PMID- 12901385 TI - Medicare program; changes to the hospital inpatient prospective payment systems and fiscal year 2004 rates. Final rule. AB - We are revising the Medicare hospital inpatient prospective payment systems (IPPS) for operating and capital costs to implement changes arising from our continuing experience with these systems. In addition, in the Addendum to this final rule, we are describing changes to the amounts and factors used to determine the rates for Medicare hospital inpatient services for operating costs and capital-related costs. These changes are applicable to discharges occurring on or after October 1, 2003. We also are setting forth rate-of-increase limits as well as policy changes for hospitals and hospital units excluded from the IPPS that are paid on a cost basis subject to these limits. Among other changes that we are making are: changes to the classification of cases to the diagnosis related groups (DRGS); changes to the long-term care (LTC)-DRGs and relative weights; the introduction of updated wage data used to compute the wage index; the approval of new technologies for add-on payments; changes to the policies governing postacute care transfers; payments to hospitals for the direct and indirect costs of graduate medical education; pass-through payments for nursing and allied health education programs; determination of hospital beds and patient days for payment adjustment purposes; and payments to critical access hospitals (CAHs). PMID- 12901386 TI - Medicare program; changes to the inpatient rehabilitation facility prospective payment system and fiscal year 2004 rates. Final rule. AB - In this final rule, we are establishing the prospective payment rates for inpatient hospital services furnished under Medicare by inpatient rehabilitation facilities (IRFs) for Federal fiscal year (FY) 2004, as required under section 1886(j)(3)(C) of the Social Security Act (the Act). As required by law and regulations, we are specifying the classification and weighting factors for the IRF case- mix groups and providing a description of the methodology and data used in computing the prospective payment rates for FY 2004. These rates are applicable to discharges occurring on or after October 1, 2003 and before October 1, 2004. In addition, we are revising and clarifying policies governing the payment for inpatient hospital services furnished by IRFs under the IRF PPS. PMID- 12901387 TI - [Bertha Lutz and the memory of Adolpho Lutz]. AB - The authors reconstruct Bertha Lutz's efforts to preserve the memory of her father, Adolpho, after his death on 6 October 1940, by studying the papers of both father and daughter that are held in the Archives of the National Museum. Bertha Lutz's plans included the construction of a museum for Adolpho Lutz's collections, the publication of his complete works and a biography. A pioneer in the feminist movement and researcher at the National Museum of Rio de Janeiro, Bertha was always a faithful assistant to her father and her own professional life followed a line of research opened by him. For more than 30 years, she dedicated herself to the consecration of her father's name, lobbying politicians and scientists in Brazil and abroad. She was not as successful as she had hoped and Bertha Lutz recorded in her will her wish that the works and biography of Adolpho Lutz be published by future generations. PMID- 12901388 TI - [Overseas disease: comparative studies of onchocerciasis in Latin America and Africa]. AB - Round worm is a parasite, Onchocera, and is transmitted by a black fly, simulidae; it can cause blindness. Originally from the African continent, where it is widespread, in Latin America it was first discovered in Guatemala in 1917; later instances were recorded in Mexico, Venezuela, Colombia and Brazil. The establishment of this disease in the Americas has intrigued scientists since then and today it is an open question. The multidisciplinary project described in this research note aims to investigate historical aspects of the arrival and spread of the disease in Latin America and, to make comparative studies of the history of the disease on both continents. The increasing importance of the disease has meant greater value being attributed to collections of simuliideo, whose Brazilian species were first studied by Adolpho Lutz at the beginning of the 20th century. PMID- 12901389 TI - [Adolpho Lutz's collection of black flies (Diptera - Simuliidae), its history and importance]. AB - This is part of a master's thesis currently being written under the auspices of the Post-Graduate Program in Animal Biology of the Federal University of Rio de Janeiro, with support from CAPES. It presents the species of black flies in Adolpho Lutz's collection, held at the Laboratory of Black Flies and Oncocercosis of the Department of Entomology of the Instituto Oswaldo Cruz. A pioneer in the study of these dipterons, Lutz described about 25 species from different places in Brazil. A vector of round worm, the black fly's importance to public health was recognized at the end of the 1920s. PMID- 12901392 TI - Medicare program; prospective payment system and consolidated billing for skilled nursing facilities--update. Final rule. AB - This final rule updates the payment rates used under the prospective payment system (PPS) for skilled nursing facilities (SNFs), for fiscal year (FY) 2004. Annual updates to the PPS rates are required by section 1888(e) of the Social Security Act (the Act), as amended by the Medicare, Medicaid, and SCHIP Balanced Budget Refinement Act of 1999 (BBRA), and the Medicare, Medicaid, and SCHIP Benefits Improvement and Protection Act of 2000 (BIPA), relating to Medicare payments and consolidated billing for SNFs. PMID- 12901391 TI - [Paths taken by Dr. Jorge Clarke Bleyer in the fields of tropical medicine and Brazilian prehistory]. AB - This research note presents a preliminary evaluation of the life history of Georg Carl Adolf Bleyer, physician, hygienist and naturalist who played a crucial role in the institution of public health in Santa Catarina and was a pioneer in tropical medicine, anthropology and archeology of Brazil. Adolpho Lutz was one of the main partners in Brazil of this equally multifacted physician who published important articles in German journals. His most thorough study was important to the history of the disciplines which which he worked; it provides the basis for understanding the conditions of medical and scientific practice in places far from the urban centers of southeastern Brazil that dominate the historiography of medicine and science. PMID- 12901393 TI - [Adolpho Lutz and the history of tropical medicine in Brazil]. AB - This text presents an evaluation of the activities of the Adolpho Lutz project and a history of tropical medicine in Brazil: progress in the preparation of texts and Lutz's correspondence for publication, the description and treatment of components of his papers, which are found at the National Museum. Appendices include a list of posts, titles, awards and decorations received by Adolpho Lutz, his bibliography and a selection of letters received by him. PMID- 12901395 TI - [The Lutzes as seen by their contemporaries]. PMID- 12901394 TI - Has bioterrorism preparedness improved public health? AB - In anticipation of future terrorist attacks, the nation has been focused on emergency preparedness, including threats to public health and the ability of communities to respond to them. The Center for Studying Health System Change's (HSC) recent site visits to 12 nationally representative communities found early benefits to public health due to heightened attention to bioterrorism preparedness: more visibility and credibility for public health, stronger public health infrastructure and improved communication and coordination across sectors. Modest negative effects included staff diversions and delays in some program implementation. As the site visits continued from fall 2002 into 2003, concerns grew that the federal smallpox vaccination program was diverting resources from such traditional public health activities as routine immunizations, health promotion and screening. PMID- 12901396 TI - Patterns of hemispheric perceptual asymmetries: left hemispatial biases predict changes in anxiety and positive affect in undergraduate women. AB - Consistent with the tripartite model of anxiety and depression, hemispheric asymmetries may be differentially associated with depressive and anxious symptoms. Indeed, research has demonstrated that asymmetries do exist when examining hemispatial biases in both anxious and depressed individuals; however, the magnitude and direction of these asymmetries has been variable. The Chimeric Faces Task was used here to measure these asymmetries, along with measures for current and future levels of anxiety and depression. Results indicated that (a) increased left hemispatial biases at Time 1 predict increased anxiety (i.e., physiological hyperarousal) at Time 2 among female undergraduate students and (b) decreased left hemispatial biases at Time 1 predict decreased positive affectivity at Time 2 among the same participants. The possibility that hemispatial biases represent a vulnerability to future anxiety and depression is discussed. PMID- 12901397 TI - Stronger suboptimal than optimal affective priming? AB - The finding of stronger affective priming in less conscious (suboptimal) conditions than in fully conscious (optimal) conditions (S. T. Murphy & R. B. Zajonc, 1993) is theoretically important because it contradicts notions that emotions are primarily reflected by conscious states. In 2 experiments, this pattern of results was obtained. Happy and angry faces were presented both optimally and suboptimally and were masked by unknown ideographs. In Experiment 1, instructions for the conscious and less conscious affective priming conditions were matched, and affective ratings of ideographs were determined. In Experiment 2, a more implicit affective measure (facial electromyography of musculus zygomaticus major and musculus corrugator supercilii) served as the dependent variable. Stronger suboptimal than optimal affective priming was found in both experiments. It is concluded that stronger suboptimal than optimal processing is characteristic for affective processing and that it can also be found when instructions are matched and when a more implicit measure is assessed. PMID- 12901399 TI - Impact of intended emotion intensity on cue utilization and decoding accuracy in vocal expression of emotion. AB - Actors vocally portrayed happiness, sadness, anger, fear, and disgust with weak and strong emotion intensity while reading brief verbal phrases aloud. The portrayals were recorded and analyzed according to 20 acoustic cues. Listeners decoded each portrayal by using forced-choice or quantitative ratings. The results showed that (a) portrayals with strong emotion intensity yielded higher decoding accuracy than portrayals with weak intensity, (b) listeners were able to decode the intensity of portrayals, (c) portrayals of the same emotion with different intensity yielded different patterns of acoustic cues, and (d) certain acoustic cues (e.g., fundamental frequency, high-frequency energy) were highly predictive of listeners' ratings of emotion intensity. It is argued that lack of control for emotion intensity may account for some of the inconsistencies in cue utilization reported in the literature. PMID- 12901398 TI - Attachment in individuals with social anxiety disorder: the relationship among adult attachment styles, social anxiety, and depression. AB - Despite their apparent implications for social functioning, adult attachment styles have never been specifically explored among persons with social anxiety disorder. In the current study, a cluster analysis of the Revised Adult Attachment Scale (N. L. Collins, 1996) revealed that 118 patients with social anxiety were best represented by anxious and secure attachment style clusters. Members of the anxious attachment cluster exhibited more severe social anxiety and avoidance, greater depression, greater impairment, and lower life satisfaction than members of the secure attachment cluster. This pattern was replicated in a separate sample of 56 patients and compared with the pattern found in 36 control participants. Social anxiety mediated the association between attachment insecurity and depression. Findings are discussed in the context of their relevance to the etiology, maintenance, and cognitive-behavioral treatment of social anxiety disorder. PMID- 12901400 TI - Emotional experience in daily life: valence, variability, and rate of change. AB - Measurements of 3 basic affective phenomena--valence of emotion, intraindividual variability, and rate of change--were derived from self-reports of emotion gathered from 123 participants (68 females, 55 males) sampled 4 times daily over 8 days. Self-reports and acquaintances' descriptions of personality were also obtained. The 3 emotion variables were not significantly intercorrelated and yielded different personality correlates. Valence of emotion positively correlated with extraversion, intraindividual variability with repression, and rate of change with fearfulness and hostility toward others, but not all results were consistent across gender. It is suggested that future research pay increased attention to patterns of emotional change over time and to the relations between gender and emotional experience. PMID- 12901401 TI - Gynecologic sonography: past, present, and future. PMID- 12901402 TI - Dance of live adult filarial worms is a reliable sign of scrotal filarial infection. AB - OBJECTIVE: To determine the value of the filarial dance sign as a diagnostic sign of scrotal filarial infection and to recognize unsuspected scrotal filariasis by this sign. METHODS: Eight symptomatic patients in whom the filarial dance sign was shown on high-resolution ultrasonography were studied, investigated, and followed after treatment with diethylcarbamazine citrate. Two patients underwent fine-needle aspiration. RESULTS: Multiple foci (nests) of motile (live) filarial worms were observed in most patients. Fine-needle aspiration of the dilated lymphatic vessels in 2 patients confirmed the presence of microfilariae. Five of 8 patients had a favorable response to treatment with diethylcarbamazine citrate. CONCLUSIONS: High-resolution ultrasonography is a useful technique for diagnosing scrotal filariasis in symptomatic patients and is very useful in the follow-up period for assessing the response of worms to treatment. PMID- 12901403 TI - Usefulness of a prevoiding transabdominal sonographic bladder scan for uroflowmetry in patients involved in clinical studies of benign prostatic hyperplasia. AB - OBJECTIVE: Flow rate measurements in clinical studies require an amount of voided urine greater than 125 to 150 mL. Often patients do not void sufficient amounts of urine, and having them produce repeated flows is often difficult. Transabdominal sonographic scanning of the bladder is a simple method for estimating bladder volume. We determined by sonography the prevoiding bladder volume needed to void a sufficient amount of urine. METHODS: Sixty-seven patients with a mean age +/- SD of 67.5 +/- 8.0 years underwent free flow rate measurement. Bladder volume was measured by transabdominal sonography when the patient had the sensation to void and after uroflowmetry to calculate residual urine. RESULTS: The voided volume was 220 +/- 127 mL; the postvoiding residual urine volume was 92 +/- 88 mL; and the bladder scan volume was 309 +/- 158 mL. Among all patients, 23.9% had insufficient voided volumes of less than 125 mL in the flow rate measurements, and 31.3% had voided volumes of less than 150 mL. There was a strong correlation between the prevoiding measured volume and the voided volume (r = 0.836; P < .0001). Linear regression analysis of the flow rate recording yielded the following formula: voided volume = 11,766 + (0.673 x prevoiding volume). CONCLUSIONS: Bladder scanning before uroflowmetry is a useful test for reducing the amount of nonevaluable flow rate data. If a voided volume of greater than 125 mL (>150 mL) is required, the mandatory prevoiding bladder scan volume should be greater than 200 mL (>250 mL), which would decrease the number of noneligible flow rate recordings from 23.9% to 4.5% (31.3% to 4.5%). PMID- 12901404 TI - Bladder wall thickness on ultrasonographic cystourethrography: affecting factors and their implications. AB - OBJECTIVE: To explore factors affecting bladder wall thickness on ultrasonographic cystourethrography in female patients with lower urinary tract symptoms. METHODS: The records of 492 female patients with lower urinary tract symptoms who had undergone a urodynamic study and ultrasonography of the lower urinary tract and who had normal urinalysis findings, negative urine culture results, or both were identified from our urogynecologic database. These included 248 patients with urodynamic stress incontinence, 38 with detrusor overactivity, 39 with mixed incontinence, 35 with a hypersensitive bladder, 42 with voiding difficulty, and 90 with normal urodynamic findings. RESULTS: Age, resting bladder neck angle, urethral mobility, and maximum urethral closure pressure were significantly associated with bladder wall thickness at the trigone and dome. Bladder wall thickness at the trigone was correlated with that at the dome (P < .0001). Bladder wall thickness at the trigone was positively correlated with pressure transmission ratios in the first and second quarters of the urethra (P < .0001; P = .002, respectively), whereas that at the dome was positively correlated with intravesical pressure at maximum flow and with detrusor opening pressure (P = .027; P = .046, respectively). Age and intravesical pressure at maximum flow were independently associated with bladder wall thickness at the trigone and dome (P = .007; P = .028), respectively. A thickened bladder wall was a common finding in female lower urinary tract symptoms, except in the patients with a hypersensitive bladder. CONCLUSIONS: Demographic, anatomic, and urodynamic factors may affect the bladder wall thickness at the trigone, dome, or both. PMID- 12901405 TI - Childhood cardiac function after prenatal diagnosis of intracardiac echogenic foci. AB - OBJECTIVE: To determine whether prenatally diagnosed intracardiac echogenic foci are associated with childhood cardiac dysfunction and persistence. METHODS: Children in whom intracardiac echogenic foci were shown on prenatal sonography at 1 perinatal center underwent echocardiography at ages 2 to 7 years. A single pediatric cardiologist, blinded to the prenatal sonographic intracardiac echogenic focus locations, assessed cardiac function by measuring the left ventricular shortening fraction and myocardial performance index. The presence of tricuspid and mitral valve regurgitation was also sought. The secondary outcome was intracardiac echogenic focus persistence. RESULTS: Twenty-five children, 14 (56%) male and 11 (44%) female, were examined at a mean age +/- SD of 3.0 +/- 1.0 years. Prenatally, 18 children (72%) had left ventricular intracardiac echogenic foci, and 7 (28%) had right ventricular intracardiac echogenic foci. The left ventricular shortening fraction was normal in all children. The overall mean left ventricular myocardial performance index (reference value, 0.36 +/- 0.06), was normal for both children with left ventricular intracardiac echogenic foci (0.36 +/- 0.06) and those with right ventricular intracardiac echogenic foci (0.36 +/- 0.04). Two children with left ventricular intracardiac echogenic foci had an isolated left ventricular myocardial performance index of greater than 2.5 SD above the mean. Trace tricuspid valve regurgitation and mitral valve regurgitation were noted in 13 (52%) and 2 (8%) of the children, respectively, similar to the general population. Left ventricular intracardiac echogenic foci persisted in 16 children (89%), whereas right ventricular intracardiac echogenic foci persisted in 2 (29%) (P = .007). CONCLUSIONS: Prenatally diagnosed intracardiac echogenic foci are often persistent but not associated with childhood myocardial dysfunction. PMID- 12901406 TI - Evaluation of contrast enhancement patterns in pancreatic tumors by coded harmonic sonographic imaging with a microbubble contrast agent. AB - OBJECTIVE: The purpose of the study was to assess patterns of primary pancreatic lesions by contrast-enhanced sonography for differentiating ductal carcinomas from other pancreatic tumors. METHODS: One hundred six consecutive patients with pancreatic masses, consisting of 83 ductal carcinomas, 7 endocrine carcinomas, 5 intraductal papillary mucinous tumors, 3 cases of autoimmune-related pancreatitis, 3 solid pseudopapillary tumors, 2 cases of chronic pancreatitis, 1 serous cystadenoma, 1 osteoclastoid giant cell tumor, and 1 follicular lymphoma, were examined by contrast-enhanced sonography with coded harmonic imaging in a phase inversion harmonic technique. The contrast enhancement patterns were assessed, and specimens removed during pancreatectomy were subjected to pathologic examination. RESULTS: Internal tumoral vascularity was detected in 47 (56.6%) of the 83 ductal carcinomas. Vascular image spreading and homogeneous staining throughout the tumors were observed in all endocrine carcinomas. Two of the 5 intraductal papillary mucinous tumors were positive for enhancement effects. Enhancement effects were observed in all 3 cases of autoimmune-related pancreatitis, but the degree varied. There was a significant correlation between the intensity of enhancement effects and the ratio of patent vessels in the tumors (P < .05). CONCLUSIONS: Vascularity was detected by contrast-enhanced sonography in only about half of the ductal carcinomas, confirming the difficulty in distinguishing those tumors from other pancreatic tumors. There was a correlation between the patency of the vessels in the tumors and their vascularity. PMID- 12901407 TI - Percutaneous sonographically guided treatment of hydatid cysts in sheep: direct injection of mebendazole and albendazole. AB - OBJECTIVE: The purpose of this study was to investigate the scolicidal effect of intracystic injection of benzimidazolic solutions in naturally infected sheep with hydatid disease. METHODS: Twenty-four sheep with 37 hydatid cysts were included in this study for percutaneous treatment with benzimidazolic solutions. The animals were divided into 3 groups: group I, treatment group with mebendazole; group II, treatment group with albendazole; and group III, control group with distilled water. All solutions were given percutaneously under sonographic guidance. Cyst contents were aspirated with a needle, and then scolicidal solutions were injected into the cysts; reaspiration was not done. Routine follow-up sonographic images were taken on the 15th day after treatment, then once per month for 3 months, and then at 3-month intervals thereafter. At the 1-month follow-up, the percutaneous aspirate yielded orange juice-like material containing necrotic debris without living scolices. RESULTS: Sonography showed a reduction in cyst size in the benzimidazolic groups (groups I and II) and progressive changes in echo patterns. An anaphylactic reaction was observed during the procedure in 1 animal. After 12 months of sonographic follow-up, the animals in all groups were killed, and macroscopic and microscopic changes in tissue samples were evaluated. At autopsy, no cysts with living scolices were found in the benzimidazolic groups, and the appearance of the treated cysts was different from that of those in the control group. Microscopic examination showed the degeneration, necrosis, and thickening of the cyst walls in the treatment groups. CONCLUSIONS: Intracystic injection of benzimidazolic solutions as scolicidal agents may be used for percutaneous treatment of hepatic hydatid cysts in sheep. PMID- 12901408 TI - Second-trimester sonographic diagnosis of diastrophic dysplasia: report of 2 index cases. AB - OBJECTIVE: To describe the sonographic features of diastrophic dysplasia in 2 second-trimester index offspring. METHODS: Real-time sonography was performed on 2 second-trimester fetuses with no risk factors for skeletal dysplasia. RESULTS: Each fetus had severe bilateral upper and lower extremity long bone shortening, hitchhiker thumbs and great toes, and clubbed feet, indicating a diagnosis of diastrophic dysplasia. CONCLUSIONS: Index cases of diastrophic dysplasia may be correctly diagnosed on the basis of second-trimester sonography. PMID- 12901409 TI - Percutaneous ultrasonographically guided thrombin injection of iatrogenic pseudoaneurysms in unusual sites. AB - OBJECTIVES: To evaluate the effectiveness and safety of percutaneous ultrasonographically guided thrombin injection as treatment of unusually positioned and unusually large iatrogenic pseudoaneurysms. METHODS: Five patients with iatrogenic pseudoaneurysms were evaluated by color duplex ultrasonography. Two patients had additional digital angiography, and 2 had additional computed tomographic angiography. In 3 of the patients, large, painful iatrogenic pseudoaneurysms located proximal (2 patients) and distal (1 patient) to the arteriovenous hemodialysis fistulas had developed, most likely due to erroneous puncture of the arterial side (brachial artery) or venous side (cephalic vein) of the fistulas. An iatrogenic pseudoaneurysm of the anterior tibial artery had developed in the fourth patient after osteotomy of the fibula, and an iatrogenic pseudoaneurysm of the superficial femoral artery had developed in the fifth patient after erroneous puncture during venous transfemoral angiography. With a sterile technique and color duplex ultrasonographic guidance, a diluted solution of bovine thrombin was slowly injected directly into the iatrogenic pseudoaneurysms until cessation of blood flow was seen. Follow-up color duplex ultrasonography was performed 24 to 48 hours after the ultrasonographically guided thrombin injection. RESULTS: Four iatrogenic pseudoaneurysms were successfully thrombosed during 1 session. Two large iatrogenic pseudoaneurysms necessitated multiple repositions of the injecting needle and several injections of small amounts of thrombin into the residual patent lumen to induce complete thrombosis without an appreciable increase in the total thrombin dosage. Follow up examinations revealed complete and persistent thrombosis without evidence of distal embolization. One iatrogenic pseudoaneurysm involving the cephalic vein, distal to an arteriovenous hemodialysis fistula, recurred after apparently successful initial thrombosis. CONCLUSIONS: Most iatrogenic pseudoaneurysms are amenable to ultrasonographically guided thrombin injection as long as they are imaged adequately by color duplex ultrasonography. PMID- 12901410 TI - Pseudoangiomatous hyperplasia of mammary stroma: sonographic appearance. PMID- 12901411 TI - Prenatal diagnosis of congenital mesoblastic nephroma in 2 siblings. PMID- 12901412 TI - Giant hemangiopericytoma of the fetal neck: prenatal two- and three-dimensional sonographic findings. PMID- 12901413 TI - In utero disappearance of the corpus callosum secondary to extensive brain injury. PMID- 12901414 TI - Prenatal sonographic diagnosis of bilateral ureteroceles: the pseudoseptated fetal bladder. PMID- 12901415 TI - Sonographic detection of emphysematous cystitis. PMID- 12901416 TI - Transvaginal sonography in the treatment of acute urinary retention due to intravesical blood clots. PMID- 12901417 TI - Sonographic detection and localization of a clinically nondetectable subcutaneous contraceptive implant. PMID- 12901418 TI - Sexually-transmitted infections in adolescents and young adults in a large city of Northern Italy: a nine-year prospective survey. AB - To determine demographic, epidemiological, clinical features and risk factors of sexually-transmitted diseases (STD) in adolescents and young adults referring to a STD centre in Northern Italy, patients diagnosed with a STD and aged 13-20 years, were prospectively evaluated from the year 1991. Teenagers showed an appreciable and increasing risk of a broad spectrum of STD, and represented 3.6 13.3% of patients yearly diagnosed with a STD in a nine-year period. More than 10% of patients were under the age of 18, and did not report condom use in the six months preceding diagnosis in 96.7% of cases. A significant increase in the female gender was found since 1993 (but our series was massively influenced by the recent immigration of female sex workers, who declared frequent condom use), while male homosexuals and drug addicts represented a minority. When excluding subjects engaged in prostitution, over 50% of patients declared only 0-1 sexual partners during the last six months, but a significant increase in sexual promiscuity was observed over time (1997-1999 versus 1991-1996) (p<.02). Over two thirds of our teenagers never used a condom prior to the diagnosis of STD, and the apparently increased temporal trend to condom utilization proved related only to the recent immigration of sex workers; lack of condom use proved significantly related to the male gender, and a lower education. Non-gonococcal STD and human Papillomavirus infection accounted for over 60% of overall diseases, while the frequency of molluscum contagiosum, gonorrhea, HSV genital ulcer, and Chlamydia disease ranged from 6.1 to 7.4%, and that of syphilis, phthiriasis, trichomoniasis, and HIV infection varied from 1.3 to 4.7%. Non-gonococcal STD and syphilis were increasingly diagnosed over time, especially in immigrant prostitutes (and despite their frequent condom use). According to our surveillance study of teenagers, a number of demographic, epidemiological, and clinical features of STD showed significant variations over time: recent immigration, spread of prostitution, increased sexual promiscuity, and infrequent condom use, are relevant and/or persisting risk factors. A permanent monitoring of STD in young adults and adolescents is strongly warranted, to allow a timely diagnosis and an appropriate treatment, and to plan preventive strategies specifically addressed to this target population, with special attention to immigrants and sex workers. PMID- 12901419 TI - The prevalence of hepatitis G virus in cancer patients. AB - Hepatitis G virus (HGV) is transfusion-transmissible and has a global distribution. However, its frequency and clinical impact have not been well established in extrahepatic cancer patients. In this study, we determined the prevalence of HGV RNA positivity, and its relationship with history of transfusion, amount of transfusion, age and sex in patients with hematological and solid malignancies in Southeastern Turkey. Sixty-one patients (35 males and 26 females) followed up for various malignancies and 60 healthy subjects were included in the study. HGV RNA was investigated by the reverse transcription polymerase chain reaction. HGV RNA was detected in none of the patients with cancer (0%) and only one person (1.6%) in the control group. There was no difference between the groups. This observation indicates that the prevalence of HGV is very low in cancer patients and healthy people in our geographical area. In addition, the role of HGV in the pathogenesis of extrahepatic malignancies seems insignificant. PMID- 12901420 TI - Genetic polymorphism by RAPD-PCR and phenotypic characteristics of isolated thermotolerant Bacillus strains from hot spring sources. AB - The polymerase chain reaction (PCR) based random amplified polymorphic DNA (RAPD) assay, morphological, physiological, biochemical and antimicrobial susceptibility test methods have been evaluated for use in the taxonomy of isolated thermotolerant Bacillus from Jordanian hot springs, with specific reference to strains Geobacillus stearothermophilus (ATCC 12980), Bacillus circulans (ATCC 4513) and Bacillus sphaericus (ATCC 14577). A RAPD assay has been optimized and is able to discriminate between numerous thermotolerant Bacillus strains. RAPD PCR was found to give reproducible thermotolerant Bacillus strains classification of DNA fingerprints for 14 strains including 3 reference strains. A study of 14 isolates and 3 reference strains, analyzing 53 phenotypic characters, resulted in their allocation to five major clusters at 60% similarity. Whereas at 80% similarity, twelve taxonomically distinct groups were evident. PMID- 12901421 TI - Prevalence of extended-spectrum beta-lactamases produced by nosocomial isolates of Enterobacteriaceae in Trakya University Hospital, Turkey. AB - The prevalence of extended-spectrum beta-lactamase (ESBL) production by 194 nosocomial isolates of Enterobacteriacea recovered from 1995 to 1999 was investigated. The ESBL production was determined by the double-disk synergy test and was confirmed by the E-test ESBL strip. Twenty-three isolates (21 Klebsiella pneumoniae, one Escherichia coli, one Providencia rettgeri) were found as ESBL producers (11.8%). These isolates were also usually resistant to non-betalactam antibiotics. Most of them contained a beta-lactamase with a pI of 7.6. All the strains conjugally transferred their ESBLs to recipient E. coli. Contrary to others, ESBL-producing K. pneumoniae strains isolated in 1999 were resistant to ciprofloxacin, and had the identical plasmid profiles suggestive of an outbreak. Ciprofloxacin resistance in these strains could not be transferred. In conclusion, K. pneumoniae was the main ESBL-producing species among nosocomial isolates of Enterobacteriacae in our hospital. PMID- 12901422 TI - Antibiotic resistance in Burkina Faso. AB - Burkina Faso is one of the Subsaharan African nations. No national services for monitoring of antibiotic resistance are available, so the number of reports of resistance patterns among hospital pathogens are inconsistent. In order to evaluate antibiotic resistance, a total of 1998 valuable microrganisms were analysed during 2000 at the Medical Centre St. Camille of Ouagadougou, Burkina Faso's capital. They were isolated as follows: 1012 from urine-culture, 503 from tonsil swabs, 398 from pus, 53 from sputum and 32 from blood-cultures. Escherichia coli was the most isolated microrganism from urine (44%); Enterococcus faecalis from tonsil swabs (96.4%), Staphylococcus aureus from pus (17%) and K. pneumoniae (70%) from sputum. In general, resistance to the old antibiotics, such as aminopenicillins and cotrimoxazole was shown. The most active antibiotic was norfloxacin, a rarely used antibiotic in this country. In conclusion, our study shows that it is necessary to create antibiotic-resistance surveillance centers in the developing countries to adopt an accurate therapy to avoid exporting of antibiotic resistance to the developed countries linked to increased emigration. PMID- 12901423 TI - Detection of listeria monocytogenes in foodstuffs using chromogenic isolation media. AB - A study was carried out to compare two selective plating media commonly used to detect Listeria monocytogenes in food samples (Palcam and Oxford agar) with two more recently formulated chromogenic media (ALOA and Rapid'L.mono). A comparison was also made between the identification systems API-Listeria and Monocytogenes ID Discs. The results obtained from the analysis of 132 food samples showed that the chromogenic media allowed Listeria monocytogenes to be detected more rapidly and with higher levels of specificity and sensitivity. The efficiency of the Monocytogenes ID Discs test was confirmed, giving results comparable to those obtained with API-Listeria, but in a simpler and quicker manner. PMID- 12901425 TI - Prevalence and chemosusceptibility of Neisseria meningitidis and Haemophilus influenzae in a population of central Italy. AB - The purpose of this study was to determine the prevalence of nasopharyngeal carriers of Neisseria meningitidis and Haemophilus influenzae, and to evaluate their chemoresistance. N. meningitidis was more frequently isolated in adolescents (10-15 years). All of meningococci were susceptible to ceftriaxone and rifampin, only one isolate showed reduced susceptibility to chloramphenicol and four strains showed reduced penicillin susceptibility. The results show that these drugs are still effective for prophylaxis and treatment in our area. All strains of H. influenzae were susceptible to penicillin, ceftriaxone, chloramphenicol, rifampin, azithromicin and gentamicin. 6 nontypeable strains were resistant to sulfamethoxazole-trimethoprim, 7 strains of type a and c-f, and 3 non-typeable strains showed reduced susceptibility to tetracycline. In contrast with the current trend in the world, in our area the susceptibility of H. influenzae to betalactams was 100%, therefore these antibiotics are still the drugs of choice for treatment of invasive diseases. PMID- 12901424 TI - Resistance emergence among P. aenrginosa in a rat thigh-abscess model. Comparison of imipenem and meropenem treatment. AB - Resistance emergence to carbapenem antibiotics was studied in a rat-thigh abscess model. Abscesses were developed in three groups with a total of 15 P. aeruginosa strains (three rats per strain). Groups were assigned to imipenem or meropenem treatment while one was left antibiotic-free. Test strains were fully susceptible to these antibiotics and the "Mutant Preventing Concentrations" of imipenem and meropenem over these strains were comparable. Antibiotic serum levels, assessed by serum bioassay test, were similar among therats. After four days, rats (n=45) were sacrificed and carbapenem resistant mutants were selected on imipenem (4 mg/L) and meropenem (4 mg/L) supplemented agar plates. Resistant variants of three strains, from four abscesses, were detected; one in the meropenem group, two in the imipenem and one in the untreated group. The MICs of imipenem and meropenem for the mutants were increased fourfold times or even higher of their counterparts. Resistance emergence under antibiotic pressure in P. aeruginosa has been shown in various conditions. To our knowledge, however, resistance emergence in abscess and also the comparison of imipenem and meropenem in this regard has not been studied before. PMID- 12901426 TI - Neutrophil superoxide anion production, and CAT and GSSGR activity in patients with tuberculosis. AB - Tuberculosis (TB) is a chronic infection disease caused by Mycobacterium tuberculosis (MTB), as an intracellular pathogen. Various cytokines (TNF-alpha, IL's, GSF etc.) and other factors play important preventing roles and are secreted during the infection. It may cause changes in the metabolism of neutrophils. Production of superoxide anion and antioxidative enzymes activities, such as glutathione reductase (GSSGR) and catalase (CAT) may be changed during MTB infection in the host. In this study, the control group consisted of ten healthy subjects and ten patients with TB were studied before anti-TB treatment. Level of superoxide anion production, activity of CAT and activity of GSSGR were studied from peripheral neutrophils of healthy subjects and patients with TB. Catalase activities of the neutrophils were significantly lower in patients with TB than normal subjects (p < 0.01). Glutathione reductase activities of the neutrophils were also significantly lower in patients with TB than normal subjects (p < 0.05). Superoxide anion production in the neutrophils did not show any significant difference between TB and normal subjects (p > 0.05). As a result, the activities of CAT and GSSGR were lower in the peripheral neutrophils of patients with TB than normal subjects, whereas superoxide anion production in the neutrophils did not differ between in TB patients than normal subjects. PMID- 12901427 TI - Nosocomial infections and risk factors in intensive care units. AB - The present study was performed to describe the incidence and risk factors for nosocomial infections (NIs) in the intensive care units of a University hospital. From January to July 2002, 454 patients from Medical and Surgical Intensive Care Unit (MICU and SICU) were observed in the active, targeted, prospective surveillance study. Risk factors for NIs were found by logistic regression analysis. Two hundred and eighteen NIs were recorded in 149 of 454 screened patients. The overall incidence rate was calculated as 33% in the ICUs. The incidence was 39% in MICU and 29% in SICU. The most frequent nosocomial infection observed in MICU, and SICU was pneumonia (42%), and surgical wound infection (31%), respectively. Extrinsic risk factors were urinary catheter, mechanical ventilation, total parenteral nutrition, entubation, antimicrobial treatment prior to the NI, nasogastric cathater and central catheter. The highest intrinsic risk factor was unconsciousness in MICU, respiratory failure in SICU. This high NI rates may be reduced by timely feedback of data for infection control activities. The recognition of risk factors for NIs is an important tool for the identification and development of interventions to minimize such risks in the ICU's. PMID- 12901428 TI - Dehydrated gelatin drops: a good method for fungi maintenance and preservation. AB - Some techniques have been proposed to maintain fungi culture collection. However, any choice must ensure the cultural stability and its phenotypic characteristics. This work proposes an adaptation of a preservation method considered by few literature reports: the dehydrated gelatin drops method (DGD). A total of 27 strains of fungi of clinical interest, including four dermatophyte fungi isolates, six filamentous non-dermatophyte fungi, five environment isolated filamentous fungi, six dimorphic fungi and six yeasts were maintained by this method for a seven year period at room temperature. After that time, the macro and micro characteristics of each fungus were studied, allowing the evaluation of the DGD method. In our experience, none of the strains maintained by DGD were found to be contaminated by bacteria or other fungi and no apparent changes were observed in morphology or macroscopic features. PMID- 12901429 TI - Children's behavioral styles at age 3 are linked to their adult personality traits at age 26. AB - We observed 1,000 3-year-old children who exhibited five temperament types: Undercontrolled, Inhibited, Confident, Reserved, and Well-adjusted. Twenty-three years later, we reexamined 96% of the children as adults, using multiple methods of comprehensive personality assessment, including both self- and informant reports. These longitudinal data provide the longest and strongest evidence to date that children's early-emerging behavioral styles can foretell their characteristic behaviors, thoughts, and feelings as adults, pointing to the foundations of the human personality in the early years of life. PMID- 12901430 TI - A multisource approach to self-critical vulnerability to depression: the moderating role of attachment. AB - The present study investigated the effects of self-criticism, dependency, and attachment variables in depression among couples. We utilized a multisource design that involved self-reports and spouse reports of personality and depression. This approach enabled us to explore the patterns of relations between self-reported and the spouse's report of the partner's view of self-criticism, dependency, and attachment dimensions, as well as the contribution of the latter to the moderation of distress. Participants were 120 couples in their first marriages. It was found that: (1) Self- and spouses' reported self-criticism are both associated with depression; (2) negative assessments of personality factors and attachment models by the self and spouse contribute uniquely in predicting depressive symptomatology; and (3) beyond the covariation between target's depression and marital maladjustment, attachment models of self and of other as reported by both the self and spouse moderate the effects of self-reported personality vulnerability on depressive symptomatology. Our results indicate that self-ratings and ratings by others must both be considered in the context of depression in close interpersonal relationships. Beyond the methodological implications of multisource data, our findings support the view of depression as an interpersonal process. PMID- 12901431 TI - Taking offense: effects of personality and teasing history on behavioral and emotional reactions to teasing. AB - This study investigated how global personality traits and teasing history are related to participants' emotional and behavioral reactions to an actual teasing event. College undergraduates (N = 108) worked on a task with a same-sex confederate. While interacting, the confederate either teased participants about how slowly they were working on the task or made a benign comment about the nature of the task. Analyses revealed that even mild teasing can generate negativity towards the teaser and interaction. More interestingly, however, personality moderated reactions to teasing, as teasing condition interacted with each of the Big Five personality domains in theoretically meaningful ways. Childhood teasing history also moderated reactions to teasing, as frequent victims and frequent teasers responded in different ways. PMID- 12901432 TI - Girls' use of defense mechanisms following peer rejection. AB - This study explores the relation between girls' social adjustment and their use of defense mechanisms. We recruited girls representing four sociometric status classifications (rejected, neglected, average, and popular), and assessed their use of defense mechanisms both before and after encountering a peer rejection experience in the laboratory. We hypothesized that increasing degrees of social maladjustment would be associated with higher levels of defense use, particularly after encountering a rejection experience. Our results supported these hypotheses. There was a significant negative relationship between social adjustment and defense use, both prior to and immediately following the rejection experience. Categorical analyses revealed that rejected and neglected girls used more defenses following the rejection experience than did popular and average girls. PMID- 12901433 TI - Personality-relationship transaction in adolescence: core versus surface personality characteristics. AB - Personality effects on perceived support from social relationships and vice versa were longitudinally studied over adolescence. Within personality, core (Big Five personality traits) and surface characteristics (global self-worth, perceived peer acceptance, and loneliness) were distinguished. Core, but not surface, characteristics at age 12 predicted support from both parents and peers at age 17 after controlling for support at age 12. Support at age 12 predicted surface, but not core, characteristics at age 17 after controlling for personality at age 12. These findings are interpreted within a dual model of personality-relationship transaction. Core characteristics are relatively stable traits that are largely immune against experiences in relationships and continuously influence their flux and flow. Surface characteristics are more open to relationship influences, and are therefore less stable. PMID- 12901435 TI - Novel poly(ethylene glycol) derivatives with carboxylic acid pendant groups: synthesis and their protection and enhancing effect on non-viral gene transfection systems. AB - Novel carboxylic acid pendant-containing poly(ethylene glycol) (PEG) derivatives (PEG-Cs) were synthesized by the chemical modification of the carbon-carbon double-bond side chain of copoly(allyl glycidyl ether/ethylene oxide) (copoly(AGE/EO)). PEG-C showed a protecting ability for the DNA/polycation complex from the albumin-induced aggregation. It also expressed the enhanced efficiency on the polycation-mediated gene transfection on the cultured cells (up to 3-4-times higher), probably due to its disperse-stabilizing property and also the proton-sponge effect. PMID- 12901434 TI - In vivo pain relief effectiveness of an analgesic-anesthetic carrying biodegradable controlled release rod systems. AB - Pain is the most common and feared symptom for patients, especially those with cancer. Treatment of chronic pain with conventional ways of medication usually fails with increasing severity of the pain. New approaches enabling the prolonged provision of pain relievers are required. We designed a controlled release system of pain relievers, mainly for opioids (morphine, M, codeine, C, and hydromorphone, HM), and a local anesthetic (bupivacaine, BP) in the form of poly(L-lactide-co-glycolide) (PLGA) rods. The efficacy of these rods implanted alone or in combination in relieving chronic pain in rats caused by the ligation of the sciatic nerve of their right hind limbs was studied. The two most common tests for measuring analgesia, i.e. tail-flick tests, that show analgesia at sites other than the site of injury, were used to study the degree of systemic distribution of the drugs and paw-withdrawal tests were used to study the analgesia at the site of injury. Alleviation of this chronic and severe neuropathic pain could be obtained for about 3-4 days when rods for two drugs, 'dual drug' (analgesic-anesthetic), were used. This duration is decreased by half (2 days) with the single-drug rods. Also the dual-drug rods, though at half the dose of each single drug application, enhanced the degree of analgesia of the first day. These in vivo results are also consistent with the previous in vitro results as in the case with codeine which had a higher first-day analgesia than morphine, despite a lower potency due to the faster in vitro release rate. Similarly, slower release of hydromorphone from PLGA (85:15) rods resulted in less systemic analgesia than the more rapidly eroding PLGA (50:50) rods of the same drug. PMID- 12901436 TI - Effect of growth factors and extracellular matrix materials on the proliferation and differentiation of microencapsulated myoblasts. AB - An alternative approach to gene therapy via non-autologous somatic gene therapy is to implant genetically-engineered cells protected from immune rejection with microcapsules to deliver a therapeutic gene product. This delivery system may be optimized by using myoblast cell lines which can undergo terminal differentiation into myotubes, thus removing the potential problems of tumorigenesis and space restriction. However, once encapsulated, myoblasts do not proliferate or differentiate well. We now report the use of extracellular matrix components and growth factors to improve these properties. Addition of matrix material collagen, merosin or laminin all stimulated myoblast proliferation, particularly when merosin and laminin were combined; however, none, except collagen, stimulated differentiation. Inclusion of basic fibroblast growth factor (bFGF) within the microcapsules in the presence of collagen stimulated proliferation of C2C12 myoblasts, as well as differentiation into myotubes. Inclusion of insulin growth factor (IGF-II) in the microcapsules had no effect on proliferation but accelerated myoblasts differentiation. When the above matrix material and growth factors were provided in combination, the use of merosin and laminin together with bFGF and IGF-II stimulated myoblast proliferation but had no effect on differentiation. In contrast, the cocktail containing bFGF, IGF-II and collagen induced increased myoblasts proliferation and subsequent differentiation. Hence, the combination of bFGF, IGF-II and collagen appears optimal in improving proliferation and differentiation in encapsulated myoblasts. PMID- 12901437 TI - Morphology and metabolism of Ba-alginate-encapsulated hepatocytes with galactosylated chitosan and poly(vinyl alcohol) as extracellular matrices. AB - Lactobionic acid, bearing a beta-galactose group, was coupled with chitosan to provide synthetic extracellular matrices together with poly(vinyl alcohol) (PVA). The hepatocytes encapsulated in Ba-alginate capsules with galactosylated chitosan (GC) and PVA as extracellular matrices showed aggregation morphologies as the incubation time increased. Ba-alginate-encapsulated hepatocytes with GC exhibited a higher metabolic function in albumin secretion compared to those entrapped in Ba-alginate beads and monolayer-cultured on a collagen-immobilized polystyrene dish. The ammonia removal ability of monolayer-cultured hepatocytes decreased with increasing culture time and disappeared completely after three days. In contrast, the ammonia removal ability of encapsulated and entrapped hepatocytes increased with increasing incubation time in the first seven and five days, respectively. Thereafter, the entrapped hepatocytes lost ammonia removal ability quickly while the encapsulated hepatocytes kept a relatively high ammonia removal ability up to 13 days. The trace amount of GC in the core matrices enabled encapsulated cells to enhance their ammonia removal and albumin secretion ability. The results obtained with 3-(3,4-dimethylthiazol-2yl)-2,5 diphenyltetrazolium bromide (MTT) also showed that the capsules incorporated with GC can provide a better microenvironment for cell aggregation along with nutrition and metabolite transfer. Due to the nature of the liquid core, the encapsulated hepatocytes showed very good mobility. This facilitated cell-cell interaction and cell-matrix interaction. PMID- 12901438 TI - Preparation of porous hydrolyzable polyrotaxane hydrogels and their erosion behavior. AB - We have prepared porous polyrotaxane hydrogels by using the salt leaching technique. Porous hydrogels were found to have a uniform and highly porous structure. The size of pores in each hydrogel was directly proportional to the size of the sodium chloride particle used. Structural uniformity of the hydrogels is useful not only for uniform cell distribution, but also for well-controlled material properties. Uniform pore size and distribution may ensure the diffusion of nutrients throughout of the gel and the removal of metabolic wastes from the system. The results of an erosion study in phosphate-buffered saline showed that the erosion time of porous polyrotaxane hydrogels was controlled by the poly(ethylene glycol) (PEG) content in the hydrogels. The erosion time of the porous polyrotaxane hydrogel was observed to be almost the same with the non porous polyrotaxane hydrogel with the same PEG content. From the erosion study, the erosion time of the polyrotaxane hydrogel may be independent of its morphology. Easy control of the erosion time in the polyrotaxane hydrogels is useful in the preparation of scaffolds for tissue engineering. PMID- 12901439 TI - Preparation of floating alginate gel beads for drug delivery to the gastric mucosa. AB - Alginate gel beads containing ethylcellulose (ALECs) were prepared and investigated with regard to buoyancy, in vitro and in vivo drug release profiles, and drug targeting specificity in the gastric mucosa. When the ethylcellulose (EC) content of ALECs containing metronidazole (MZ) was higher than 3%, the beads floated in all test solutions with a specific gravity of approx. 1.01. ALECs containing 5% EC released MZ gradually and floated throughout the experimental period in simulated gastric juice (pH 1.2), and all of the drug had been released after 90 min. When we orally administered ALECs to guinea pigs, about 85% of the incorporated MZ was released at 1 h. The MZ concentration of the gastric mucosa after administration of ALECs was greater than that observed with administration of MZ solution, despite lower serum concentrations. Furthermore, the similar data were obtained for ALECs with 7% EC. These results suggest that ALECs may become a practical vehicle for delivering drugs to the gastric mucosa. PMID- 12901441 TI - Effect of synthesis temperature of PEO-grafted PU/PS IPNs on surface morphology and in vitro blood compatibility. AB - When hydrophilic/hydrophobic polymers have a microdomain structure, platelet adhesion and activation are effectively suppressed by prohibition of the excessive assembly of glycoproteins and adenosine triphosphate (ATP) consumption of the platelets on the surface. In this study, poly(ethylene oxide)-grafted hydrophilic polyurethane (PU)/hydrophobic polystyrene (PS) interpenetrating polymer networks (IPNs) were synthesized by varying the synthesis temperature to control the phase separation and the microdomain surface structure, and the effect of the degree of phase separation on the in vitro blood compatibility. The size of the dispersed PS-rich domains in the PU-rich matrix decreased, and the hydrophilicity also decreased as the synthesis temperature of the PS network during the IPN synthesis was decreased, as the phase separation was suppressed during the synthesis. The amount of the adsorbed bovine plasma fibrinogens (BPF) on the PEO-grafted PU/PS IPNs decreased as the synthesis temperature was decreased, and the in vitro adhesion of the platelets was also suppressed on the PEO-grafted PU/PS IPNs prepared at lower temperature. The microdomain structure on the surface affected the adhesion and the activation of the adhered platelets, and the suppression of the phase separation resulted in the decrease of the domain size, which also enhanced the blood compatibility of the PEO-grafted PU/PS IPNs. PMID- 12901440 TI - BHK cell attachment and growth on EDA-plasma-modified poly(L-lactide/epsilon caprolactone) biodegradable films. AB - In this study, attachment and growth of Baby Hamster Kidney (BHK) cells on ethylene diamine (EDA)-plasma-treated poly(L-lactide/epsilon-caprolactone) biodegradable copolymer films were investigated. The co-polymer (Mw: 58000; Mn: 35000 and PI 1.60) was synthesised by ring-opening polymerization of the respective dimers with using stannous octoate as the catalyst. The final ratio of L-lactide to epsilon-caprolactone obtained by 1H-NMR was 87:13. The co-polymer films were treated with the EDA-plasma in a glow-discharge apparatus. The BHK-30 cell line was cultured on plain and EDA-plasma-treated films and their pre-wetted forms (with ethanol and/or cell culture medium before use). Cell attachment and growth were followed. Alkaline phosphatase (ALP) activity and glucose uptake in cell culture medium were also investigated. There was no attachment in the first 12 h. Glow-discharge treatment increased significantly the attachment and growth. Pre-wetting with ethanol and cell culture medium was also increase significantly both the attachment and growth. PMID- 12901442 TI - Evaluation and refinement of the environmental stress index for different climatic conditions. AB - The purpose of this study was to evaluate the recently constructed environmental stress index (ESI) for a large database comprising various climatic conditions. Data analysis of measurements from 19 locations revealed a high correlation between ESI and the wet bulb globe temperature (WBGT) index for each database. Validity from statistical analysis, including optimization procedures, slightly changed the ESI constants as follows: ESI = 0.62Ta - 0.007RH + 0.002SR + 0.0043(Ta x RH) - 0.078(0.1 + SR)(-1) where: Ta = ambient temperature (degrees C); RH = relative humidity (%); and SR = solar radiation (w x m(-2)). The refined ESI and the WBGT index were applied to databases of more than 125,000 measurements for each variable: Ta, RH, SR, black globe temperature (Tg), and wet bulb temperature (Tw). For each database, the ESI was then successfully correlated with the WBGT (P < 0.05, R2 > or = 0.899). We conclude that the refined ESI, which is constructed from fast response and commonly used weather sensors (Ta, RH, SR), is a potential index to serve as an alternative to the WBGT for heat category assessment. PMID- 12901443 TI - Combined environmental stress and physiological strain indices for physical training guidelines. AB - The purpose of this study was to develop guidelines based on the previously described environmental stress index (ESI) and physiological strain index (PSI) for work-rest cycles (WRC) during training, especially in the military. The ESI was introduced as a potential substitute for the Wet Bulb Globe Temperature Index because of the very high correlation coefficients between them. The ESI is constructed from the fast-reading meteorological response sensors ambient temperature (Ta), relative humidity (RH), and global radiation (GR), which require only a few seconds to reach equilibrium. The ESI, the first stress index using direct measurements of solar radiation (SR), is calculated as follows: ESI = 0.63Ta - 0.03RH + 0.002SR + 0.0054 (Ta x RH) - 0.073(0.1 + SR)(-1). The PSI is based on heart rate (HR) and rectal temperature (Tre) and can indicate heat strain online and analyze databases. The PSI is constructed as follows: PSI = 5(Tret - Tre0) x (39.5 - Tre0)(-1) + 5(HRt - HR0) x (180 - HR0)(-1), where Tre0 and HR0 are the initial Tre and HR, and Tret and HRt are simultaneous measurements taken at any time. The PSI is scaled from 0 to 10, whereby the respective variables, PSIHR and PSITre, representing the cardiovascular and thermoregulation systems, can contribute up to five units to the overall strain assessment. To integrate the PSI and ESI, we decided to use only the PSIHR component, which represents the metabolic rate and the strain reflected by the cardiovascular system. Furthermore, PSIHR is easier to measure, is easier to implement, and simplifies the integration with ESI. Concomitantly, PSIHR categorizes the strain between 0 and 5, the higher the value, the higher the strain. We believe that the use of the PSI WRC values will help in decreasing the risk of future heat injuries. PMID- 12901444 TI - Lipid peroxidation and antioxidants in plasma and red blood cells from patients with pemphigus vulgaris. AB - In pemphigus vulgaris, the increased production of reactive oxygen species from activated neutrophils decreases concentrations of antioxidant vitamins and enzymes in plasma and red blood cells (RBC), resulting in oxidative stress. We compared lipid peroxidation, a measure of reactive oxygen species production, antioxidant vitamins, reduced glutathione (GSH), glutathione peroxide (GSH-Px), and catalase enzyme activity in blood samples obtained from 18 nonsmoking pemphigus vulgaris patients and an equal number of age- and gender-matched, healthy control subjects. Plasma and RBC lipid peroxidation levels (malonyl dialdehyde) were significantly higher (p < 0.05) in pemphigus vulgaris patients than in control subjects. Significantly lower concentrations of plasma antioxidant vitamins (vitamin E and beta-carotene) and vitamin A (p < 0.001), antioxidant enzymes (catalase in RBC and plasma, GSH-Px in RBC [p < 0.05]), and respective GSH activities in both RBC and plasma (p < 0.05 and p < 0.01) were found in pemphigus vulgaris patients than in control subjects. GSH-Px in plasma did not change significantly. The results provide evidence for a potential role of increased lipid peroxidation and peroxidation and decreased antioxidants in pemphigus vulgaris by its inflammatory character. PMID- 12901445 TI - Evaluation of dimethoate-induced implantation delay and nidation by progesterone in albino mice. AB - We investigated the effect of dimethoate, an organophosphorus insecticide, and progesterone on implantation. Virgin pregnant albino mice received dimethoate orally at a dose of 28 mg/kg body wt/d from days 1 to 7. Laparotomy on day 8 showed no implantation sites. Thereafter, graded doses of progesterone, 4, 9, and 12 mg/kg body wt/d, were administered up to day 15. A group of control mice received a similar quantity of distilled water. Autopsy on day 8 revealed that the control mice were pregnant, with a normal number of implantations and 8.08% pre-implantation loss, whereas treatment with dimethoate for 7 days or with dimethoate for 7 days followed by progesterone for 8 days totally abolished implantation, with a 100% pre-implantation loss. In all treated mice, a significant decrease occurred in body weight gain, as well as in the weight of the ovaries, uterus, and liver when compared with those of control mice. No significant changes were found in other organ weights (kidneys, adrenals, spleen, thymus, or thyroid). The observed effect of dimethoate could be due to an imbalance in the estrogen-progesterone ratio essential for implantation. Alternatively, dimethoate treatment could result in blastotoxicity or have an impact on the hypothalamic-pituitary axis. PMID- 12901446 TI - Involvement of cholinoceptors in cadmium-induced endothelial dysfunction. AB - Cadmium (Cd) toxicity was produced in male rats to study the role of cholinoceptors in Cd-induced endothelial dysfunction. The changes in the tension of the aortic rings to constrictor and dilator agonists were compared with those of controls. A Cd-induced significant increase in phenylephrine response was associated with a decrease in basal dilator prostanoid release. In Cd-exposed rings, despite an obvious depression in the acetylcholine (ACh) response, the receptor-independent dilation to the calcium ionophore A23187, which elicits a receptor-independent endothelial relaxation, was slightly elevated (p<0.01), but the smooth muscle cell response to the NO donor, sodium nitroprusside (SNP) remained unaltered. Cadmium decreased both the maximal response to ACh (10(-5) M) and its pirenzepine (Prz) sensitive component. The M1 type cholinoceptor-mediated response to ACh decreased in Cd-exposed rings to 10.30 +/- 5.00% from 38.40 +/- 6.90% (p<0.001). Cadmium also reduced the share of indomethacin 1.64% to 13.92 +/ 2.89% (p<0.01), which correlated well with the changes in the M1-mediated response (r=0.991, p<0.0001). Most of the deleterious effect of Cd appears to be restricted to the M1-dependent ACh response. These findings suggest that Cd produces an endothelial dysfunction by impairing the M1 type cholinoceptor mediated response, which seems to be involved in prostanoid release. PMID- 12901447 TI - Comparison of consumption effects of brief interventions for hazardous drinking elderly. AB - We sought to determine if Brief Interventions [BIs, Motivational Enhancement (ME), and Brief Advice (BA)] reduced alcohol consumption among hazardous alcohol drinking elderly (65 years or older) and whether the elderly responded similarly to younger populations. In 12 primary care offices from October 1995 to December 1997, we screened 13,438 patients of whom 2702 were elderly (180 were hazardous drinkers). Forty-five elderly enrollees were randomized to receive ME (n = 18), BA (n = 12), and Standard Care (SC, n = 12). At baseline, the elderly drank more alcohol and abstained fewer days than the younger cohort (p < 0.05). During the year, the elderly in ME, BA, and SC intervention arms increased the number of days abstained, decreased the number of drinks per day, and reduced the number of total days per month drinking. There were trends toward decreases in the alcohol consumption measures in the ME and BA treatment arms compared to SC. The elderly's response to all interventions was similar to that of the younger cohort. This study suggests that hazardous alcohol consumption in the elderly is common and that BIs reduce alcohol consumption in the elderly similar to younger populations. PMID- 12901448 TI - Family attention and tobacco smoking among adolescents in Central America, Panama, and the Dominican Republic. AB - This study examined the association between family attention and tobacco use among 5549 adolescent students in five Central American countries, Panama, and the Dominican Republic who participated in a survey of drug use in 1994. Drug use and other variables were assessed using an adapted version of the Drug Use Screening Inventory (DUSI) in Spanish. Students with the highest level of family attention had a lower occurrence of tobacco smoking than students with the lowest level of family attention. Country-specific analyses show similar associations. These findings underscore the need to understand tobacco use in Central America and neighboring countries and to test whether interventions aimed at enhancing parental-child attention, communication, and monitoring reduce the incidence of tobacco use among youths. PMID- 12901449 TI - The impact of smoking and other substance use by urban women on the birthweight of their infants. AB - The impact of maternal smoking and other substance use during pregnancy on infant birthweight is demonstrated in a sample of 766 urban women, using data collected in the Washington, D.C. Metropolitan Area Drug Study (DC*MADS). Women residing and giving birth in the District of Columbia were interviewed in 1992. A multivariable linear regression model was used to quantify the association between birthweight and the mother's use of cigarettes, alcohol, or illicit drugs during pregnancy, while controlling for possible confounding variables. The analysis focused on factors, including prenatal care and substance use during pregnancy that may contribute to low birthweight infants born to this sample of urban, predominantly black women. A woman's use of cigarettes, marijuana, and heroin during pregnancy was related to infant birthweight, but her use of alcohol and cocaine during pregnancy was not significantly related. Smoking during pregnancy was a strong predictor for low birthweight, suggesting that targeting more smoking cessation programs for pregnant women, particularly those who may also be illicit drug users, could help reduce adverse health consequences for low birthweight infants. PMID- 12901450 TI - Health and performance related reasons for wanting to quit: gender differences among teen smokers. AB - Adolescents frequently state health as a broad-ranging reason for wanting to quit smoking. Much less is known regarding performance-related reasons. We hypothesized that more male than female smokers want to quit for performance related reasons (e.g., to improve athletic performance). As part of a telephone screen to determine eligibility for participation in a cessation trial in Baltimore, Maryland, 1999-2001, 509 teenage smokers [mean age 15.78 +/- 1.65 years (range 11-21), 60.9% female, 32.6% African-American] were asked the open ended question: "Why do you want to quit?" Responses were subsequently grouped into categories that included health, performance, cost, social influences, setting an example for others, self-efficacy, cosmetics, no perceived positive reinforcement, or unknown reasons. Health was the most commonly stated primary and overall reason for wanting to quit among both boys and girls. Sixty-five percent of teen smokers endorsing health reasons were girls, and 51% of those endorsing performance-related reasons were boys (chi2(2) = 7.78, p = 0.02). Recognizing the greater concern for performance-related issues among boys is important for designing and engaging young smokers into cessation interventions. PMID- 12901451 TI - Parental status of women injection drug users and entry to methadone maintenance. AB - This article examines patterns of methadone maintenance treatment entry among 9018 adult women injection drug users (IDUs), with special attention to parental status differences. The data originate from a statewide drug-treatment database covering all women IDUs who entered drug treatment in the State of Massachusetts over a four-year period. Through the use of logistic regression analysis, the study found that among these women IDUs, those who resided with their children were significantly more likely to enter methadone maintenance than women who were mothers but did not reside with their children. Mothers residing with their children were 73% more likely to enter methadone maintenance than mothers who do not reside with their children. The authors discuss specific programmatic and policy implications including the need to determine whether methadone maintenance should be promoted as an alternative drug treatment option in order to preserve family unification or promote family reunification. PMID- 12901452 TI - Are Hispana IDUs Tecatas?: reconsidering gender and culture in Hispana injection drug use. AB - The roles of gender and culture remain elusive in the discussion of minority male vs. female Injecting Drug User (IDU) populations. A case in point is that of Hispano and Hispana IDUs. The commonly used street term for the Hispano (male) IDU, Tecato, is feminized through the use of the corresponding female noun Tecata. To do so however, minimizes the intricacies of gender and culture in the male vs. female roles of Hispano and Hispana IDUs (e.g., daughter/son, mother/father, partner, citizen). Hispano IDUs are also impacted by gender and culture in that Hispanos who embrace the Tecato way of life, must sacrifice and inure the consequences in how their male roles are minimized (e.g., the absent male figure in community, home, and family life). This article explores, through the analysis of qualitative interview data, the demands of gender and culture as they play themselves out in the lives of Hispana IDUs. Findings from this study suggest there are important gender and culture differences between Hispano and Hispana IDUs. Implications of this study include the need to reexamine the roles of gender and culture as they cause added pressure to Hispana IDUs (i.e., in demands to fulfill gender and culture stereotyped roles while also pursuing an IDU career). PMID- 12901453 TI - Family harmony as a protective factor against adolescent tobacco and alcohol use in Wuhan, China. AB - PURPOSE: To investigate the association between family harmony (FH) and tobacco and alcohol use (TAU) in Chinese adolescents. METHODS: Participants completed a survey in 1998 as part of a larger study of adolescent health in Wuhan, China. Analyses were performed on subjects for whom complete data were available (n = 183; 50.8% male; mean age = 13.17 yrs, std dev = 0.59). Structural equation modeling was utilized to quantify the relationships between the FH, TAU, depression, and academic aptitude factors. RESULTS: The conceptualized structural equation model was found to have a good fit to the data (CFI = 0.995; chi2 = 39.57 df = 38; p = 40). FH was a significant predictor of TAU (beta = -0.42, p < 0.05) and was protective. FH' was also negatively related to depression (r = 0.24, p < 0.05) and positively related to academic achievement/aptitude (r = 0.35, p < 0.05). CONCLUSIONS: These central findings highlight the value and importance placed on FH within the Chinese culture. Future prevention programs may benefit by taking into account FH as a potential mediator of TAU in adolescents in China. PMID- 12901454 TI - Club drugs and sex on drugs are associated with different motivations for gay circuit party attendance in men. AB - Data were collected from 1169 gay circuit party patrons in three North American circuit parties in three geographical locations in 1999. Questionnaires obtained reasons for attending circuit parties and drugs used at circuit parties. Factor analyses of the 10 major reasons for attending parties indicated that there were two significant moderately correlated dimensions: a social and celebratory one to be with friends and dance, and a sensation-seeking one, to have sex and drugs. Drug and sex on drugs predictors of the social dimension accounted for only 3.8% of its variance and included alcohol, ecstasy (methylenedioxymethamphetamine), GHB (gamma-hydroxybutyrate), and having sex while on alcohol as significant predictors. Drug predictors for the sensation-seeking dimension accounted for four times the variance of the first dimension and included poppers, alcohol, ecstasy, Special K, and GHB, and having sex while on methamphetamines and on GHB. Unsafe sex was significantly associated with the sensation-seeking dimension but not the social dimension. While 63% checked > or = 3 of the social reasons for attendance, only 13% checked > or = 3 of the sensation-seeking reasons for attendance. Age was significantly inversely associated with the social dimension. These data suggest that while drug use is significantly associated with both the sensation-seeking and social dimensions of circuit party attendance, a greater number of drugs, sexual activity while on drugs, and unsafe sex are more closely associated with the sensation-seeking dimension of attendance. PMID- 12901455 TI - The word support is a paradoxical concept. PMID- 12901456 TI - The AMSUS History of Military Medicine Essay Award. The story of the mobile army surgical hospital[corrected]. AB - The critical task of the physician uniting effective surgical intervention within close proximity to the front lines, and therefore the wounded, must be matched with the challenge of not becoming a casualty as well. The U.S. Army Medical Services during the Korean War balanced the surgeon's ability to protect himself and save others. This article tells the story of perhaps one of the most characteristically highlighted medical aspects of the Korean War, the Mobile Army Surgical Hospital. Originally intended to be close to the front lines of fighting, the Mobile Army Surgical Hospital was equipped to move on its own. Initially, the unit was formed to provide surgical capabilities for one division. It transformed through the course of the war into a multidivision and multinational all-purpose hospital. With this expansion in workload (medical cases in addition to surgery) without an equitable increase in personnel, rapid evacuation of patients was the only means of keeping up. PMID- 12901457 TI - The Federal Nursing Services Award. Enlisted women with breast cancer: balancing demands and expectations. AB - Diagnosis with breast cancer is not an automatic cause for discharge from the military. Therefore, it is important to know how this disease impacts enlisted women in the military. This study describes how enlisted women manage diagnosis and treatment within the context of their military careers. Grounded theory guided the study methodology. Audiotaped semistructured interviews were conducted three times over 9 months to learn how treatment impacted work demands. Data saturation was attained after 46 interviews were conducted. Participants had to balance demands and expectations among the Military Career and Military Medical Subsystems and their Social Support Subsystem. Support from the chain of command was critical in women's ability to balance demands and expectations of career, family, and illness. Military nurses are in a unique position to create strategies that will assist enlisted women in coping with breast cancer. PMID- 12901458 TI - Anthrax refusers: a 2nd infantry division perspective. AB - The Department of Defense anthrax vaccination program has been in the news often recently. Concerns are cited over the safety and usefulness of the vaccine. This brief report describes some of the characteristics of anthrax vaccine refusers. This report examines the implementation of an anthrax vaccination program in a well-disciplined, forward-deployed Army unit facing a hostile enemy with access to anthrax biological warfare stocks. PMID- 12901459 TI - Tuberculosis outbreak investigation of a U.S. Navy amphibious ship crew and the Marine expeditionary unit aboard, 1998. AB - A Marine deployed aboard a U.S. Navy amphibious ship had smear-positive, cavitary pulmonary tuberculosis (TB). Contact investigation ultimately found 21 active cases of TB among sailors and Marines who were aboard the affected ship. Approximately 3 months lapsed between onset of the source patient's illness and appropriate diagnosis and treatment. During the contact investigation, 3,338 persons received tuberculin skin tests and 712 were identified as new latent tuberculosis infection cases. Four persons diagnosed with latent tuberculosis infection developed active TB because of poor compliance with treatment. After personnel disembarked from the ship, persistent efforts to identify persons with active disease and latent infections were successful in controlling further spread of tuberculosis in military units and local communities. The Mycobacterium tuberculosis bacteria isolated from the source patient and 16 of the other active cases were susceptible to all drugs commonly used to treat TB. PMID- 12901460 TI - A retrospective study evaluating the use of the panoramic radiograph in endodontics. AB - Patients are referred to the endodontist to have root canal therapy performed to treat pulpal and periradicular diseases. Routinely, the only radiograph to accompany the patient is the periapical radiograph. This radiograph is inadequate in the detection of asymptomatic pathosis that may be present in other areas of the maxilla and mandible. The military's readiness mission requires that a panoramic radiograph be part of the patient's dental record. In addition to its use for personal identification purposes, the panoramic radiograph is an excellent diagnostic tool that can give the clinician an overall view of the dentoalveolar structures. This retrospective study evaluated randomly selected panoramic radiographs and recorded the presence of radiolucent and radiopaque areas not evident on a referral periapical radiograph. The results of this study found a 4.2% occurrence of undiagnosed pathosis following additional radiographs and clinical examinations. PMID- 12901461 TI - A comparative analysis of amlodipine and felodipine in a military outpatient population: efficacy, outcomes, and cost considerations. AB - BACKGROUND: This retrospective study compared the efficacy, tolerability, and cost of two dihydropyridine calcium channel blockers. METHODS: Charts of patients who had been on continuous antihypertensive therapy with amlodipine or felodipine for at least 6 months were reviewed. Analyses include mean changes in blood pressure, percentage of patients achieving blood pressure (BP) < 140/90 mm Hg, average dose, and cost per day of the two calcium channel blockers, average cost of additional medication, total medication cost per day, and cost to achieve BP control. RESULTS: Eighty-seven percent of amlodipine-treated patients achieved BP control compared with 33% of felodipine-treated patients. Total medication cost to achieve BP control was 0.87 dollars per day for patients on amlodipine compared with 1.79 dollars per day for patients on felodipine. CONCLUSIONS: Amlodipine produced BP control in a greater percentage of patients than did felodipine at a lower total cost to achieve BP control. When evaluating the total cost of antihypertensive treatment, the cost of a drug alone can be misleading. PMID- 12901462 TI - Treatment of lower extremity injuries due to antipersonnel mines: blast resuscitation and victim assistance team experiences in Cambodia. AB - The two teams comprising the fiscal year 2001 Blast Resuscitation and Victim Assistance mission had the opportunity to learn from and practice mine injury treatment principles with experienced local and international war surgeons in Cambodia. Treatment principles were modifications of International Committee of the Red Cross recommendations. A total of 14 acute lower extremity mine injuries were treated. Surgery generally consisted of an open amputation or thorough irrigation and debridement using equipment readily available in any U.S. military field hospital. The surgical techniques will be described in detail. Delayed primary closure occurred 5 days later followed by prosthesis fitting (for amputees) in an International Committee of the Red Cross facility 12 weeks later. Other options and techniques will be discussed with an emphasis on applicability to U.S. military field surgery. PMID- 12901463 TI - Emergency cricothyrotomy. AB - Emergency cricothyrotomy is a potentially lifesaving surgical procedure used to gain prompt access to an otherwise compromised and inaccessible airway. The purpose of this photoessay is to demonstrate the technique of the procedure in a step-by-step manner so that the physician can perform this intervention with ease and facility in the most stressful of circumstances. PMID- 12901464 TI - Do spinal cord injuries adversely affect serum lipoprotein profiles? AB - OBJECTIVES: Previous reports indicate that dyslipidemia is very common in persons with spinal cord injury. This study aimed to examine the relationship between spinal cord injury and serum lipoprotein profiles. METHODS: We assessed 28 healthy control subjects matched for age and sex and 60 persons (46 men and 14 women; mean age, 28.1 years) with spinal cord injury having a mean duration of injury of 3.6 +/- 0.8 years (range, 1-13 years). RESULTS: Serum low-density lipoprotein cholesterol level was higher (109 +/- 37, 91 +/- 23.7 mg/dL, respectively) (p = 0.04) and serum high-density lipoprotein cholesterol level was lower (33 +/- 7.1, 44 +/- 7.4 mg/dL, respectively) (p < 0.001) in the patients with spinal cord injury than in the control. Apolipoprotein A-I level was significantly low (94 +/- 10.3, 103 +/- 11.5 mg/dl) (p = 0.003) and apolipoprotein B level was significantly high (93 +/- 15.6, 81 +/- 13.7mg/dL) (p = 0.004) in the group of spinal cord injury. No significant difference was observed the other parameters of these two groups. CONCLUSION: Our results have shown that serum lipoprotein level should not be ignored for the follow-up of the patients with spinal cord injury. PMID- 12901465 TI - Dental class 3 intercept clinic: a model for treating dental class 3 soldiers. AB - OBJECTIVE: Dental Fitness Classification (DFC) 3 indicates that a soldier has a dental condition that, if not treated, is expected to result in a dental emergency within 12 months. Forty-two percent of Army recruits have DFC 3 conditions as reported by the 2000 Tri-Service Center Recruit Study. Initial entry training schools allow minimal time for dental care. The Fort Hood Class Three Intercept Clinic (CTIC) was created to treat DFC 3 conditions prior to soldiers being assigned to their units. The purpose of this study was to evaluate CTIC's ability to make soldiers dentally deployable before joining their new units at Fort Hood, Texas. MATERIALS AND METHODS: A retrospective review of the CTIC daily sign-in logs for the period of January 1999 to May 2001 was used to identify DFC 3 soldiers. The data were analyzed to determine frequency of DFC 3 by rank, DFC after CTIC visits, distribution of treatment type needed to establish dental readiness, and to identify soldiers whose DFC 3 conditions were not treated. Additionally, for the 6 months between November 2000 and April 2001, all soldiers who remained a DFC 3 when released to their units were followed to assess how long it took for them to attain a DFC 1 or 2 status. RESULTS: The retrospective review of the CTIC records identified 5,851 DFC 3 soldiers who in processed to Fort Hood from January 1999 to May 2001. CTIC successfully converted 86.9% (N = 5,083) of the soldiers to DFC 1 or 2. Of a sample of 185 DFC 3 soldiers released to their units as a DFC 3, only 54% had converted to DFC 1 or 2, with 60.5 days being the mean days to convert. CONCLUSIONS: Trainees have large demands for dental care with minimal time to seek treatment during Initial Entry Training. A CTIC, or hybrid, can provide the necessary emergent dental care to remove soldiers from DFC 3 prior to their being assigned to their first permanent duty station. PMID- 12901466 TI - A comparative study of panretinal photocoagulation and vitrectomy for advanced diabetic retinopathy. AB - A retrospective, cohort study of patients with diabetic retinopathy was conducted, which assessed factors of age at the time of initial panretinal photocoagulation, sex, race [American Indian and non-Indian), and those requiring postpanretinal photocoagulation vitrectomy. Frequency analyses were used to categorize the study population by ethnicity and sex. Independent sample Student's t test were used to compare means of age, ethnicity, and sex. Layered chi2 analyses were performed to study the ethnic status of the men and women requiring vitrectomy, followed by an examination of the relationship of these three factors to the vitrectomy group using a logistic regression model. Within this 265 patient group, the American Indian male was noted to require vitrectomy more frequently and tended to receive initial panretinal phototcoagulation at a lower age. PMID- 12901467 TI - Does point-of-purchase nutrition labeling influence meal selections? A test in an Army cafeteria. AB - OBJECTIVE: This study assessed the effectiveness of nutrition labeling on sales of targeted entrees and measured the perceived influence that factors such as taste, quality, appearance, fat content, calorie content, and price had on meal selection behavior within an Army cafeteria. METHODS: A quasi-experimental design was used to compare targeted entree sales between a 1-year baseline period and two 30-day postintervention periods, after the placement of entree nutrition labels. A brief questionnaire, distributed to 149 patrons, measured the perceived influence of the aforementioned factors on selections. RESULTS: Analysis of variance detected no significant differences in sales between baseline and the two intervention periods; the factors of taste and quality were rated most influential to meal selection (p < 0.000). CONCLUSIONS: A marketing campaign focusing on the health attributes of targeted entree items was not successful in boosting sales. Sensory attributes (i.e., taste, quality, and appearance) appear to be more influential to meal selection. PMID- 12901468 TI - Pseudofolliculitis barbae: review and update on new treatment modalities. AB - Pseudofolliculits barbae, PFB, is a common cutaneous disease encountered frequently in medical practice. PFB represents a chronic inflammatory condition of the hair follicle caused by ingrown hairs producing an inflammatory foreign body reaction. The pathogenesis of PFB is multifactorial. Factors such as hair type and direction of hair growth play a role in the initial inflammatory reaction. In the armed forces, PFB represents a real challenge for both the physician and the patient. The combat environment, with the recent threat of biological and chemical weapons, requires the servicemen to be clean-shaven for appropriate gas mask fitting around the face. This article will review the etiology, pathogenesis, classification, and newer treatment modalities in the management of PFB. PMID- 12901469 TI - Pattern of viral hepatitis infection in a selected population from Saudi Arabia. AB - Viral hepatitis is an important health problem worldwide. Globally, three major viruses are the leading cause of hepatitis: hepatitis A, B, and C. In this study, we have investigated the pattern of hepatitis among the National Guard personnel and their extended families seen in the central region of Saudi Arabia. The most dominant type of hepatitis infection was hepatitis B virus (HBV), followed by hepatitis C virus (HCV), and to a lesser extent hepatitis A virus (HAV). Our results showed three different age groups at risk of acquiring the infection: HAV, which is mainly a disease of the young; HBV, a disease of adolescents and adults; and HCV, a disease of the elderly. There was no significant difference in the male:female ratio in HAV and HCV; however, as seen in the developed countries, more males were affected with HBV than females. These data will pave the way for further studies by identifying the risk groups for the three major hepatitis infections and by using the data as a preventive tool to educate those risk groups. PMID- 12901470 TI - Military women's risk factors for and symptoms of genitourinary infections during deployment. AB - Symptoms of vaginitis and urinary tract infections are miserable, distracting, and significantly affect women's quality of life. Among civilian women, these symptoms account for 10.5 million office visits per year. To examine the scope of the problem for military women during deployment situations, surveys were sent to randomly selected Army and Navy units. Of 841 women who completed the anonymous survey and had been deployed, vaginal infections were experienced by 30.1% and urinary tract infections by 18.4% of them during deployment. Vaginal symptoms were consistent with symptoms associated with the three most common vaginal infections (candida, bacterial, and trichomonas vaginitis). A variety of risk factors, both behavioral and situational, significantly differentiated women with and without infections. Urinary tract infections and vaginal infections are common during deployment situations where resources for self-care and appropriate primary health care for women are scarce or unavailable. One solution is a self diagnosis and treatment kit for deployed military women. PMID- 12901471 TI - The epidemiology of personality disorders in the U.S. Navy. AB - This study evaluated the incidence of first hospitalizations for personality disorders and their association with career outcomes for Navy enlisted personnel. Gender differences, rates for subtypes, and changes in hospital admission rates over time also were examined. Personnel who were first hospitalized with a personality disorder between 1980 and 1988 were followed through 1992 and were compared with a control group that did not manifest such pathology. Hospitalized personality disorder cases demonstrated pathology very early in their military careers, which was judged to have been present prior to enlistment in more than one-half of the cases. Women were more likely than men to have a first hospitalization for personality disorders. Rates in both men and women increased during the latter one-half of the 1980s and decreased during the early 1990s. PMID- 12901472 TI - Childhood obesity's relationship to time spent in sedentary behavior. AB - OBJECTIVE: The purpose of this study was to evaluate various types of sedentary behavior children participate in and to look for an association to childhood obesity. METHODS: Questionnaires were used to gather data on physical and sedentary activity, dietary intake, demographics, and anthropometrics of 9- to 12 year-old military dependents and their parents. RESULTS: Using body mass index (BMI), 39.8% of children were obese. A significant relationship was found between childhood obesity and computer usage, television watching, total hours in sedentary behavior, and maternal BMI. An indirect significant relationship with childhood obesity was also shown if a parent was home when the child got home from school and if a father participated in exercise with their child. Caloric intake, total time in physical activity, demographic variables, and father's BMI showed no significant relationship with children's BMI. CONCLUSION: Interventions should be designed targeting total time spent on the computer, total time watching television, and maternal obesity in child obesity programs. PMID- 12901474 TI - Delays in diagnosing cancer. Threat to the patient-physician relationship. PMID- 12901473 TI - Dyspnea on exertion, fatigue, and pallor in a 50-year-old active duty soldier. AB - OBJECTIVE: To discuss a comprehensive diagnostic approach to an active duty patient presenting with dyspnea on exertion, fatigue, and pallor. A 50-year-old active duty E-6 white male in Al Udeid, Qatar, presented with progressive dyspnea on exertion, fatigue, and new pallor. This case illustrates the course of events from Al Udeid to the Walter Reed Army Medical Center, where the final diagnosis was made. Along the way, questions with discussions explore the various diagnostic and management aspects of his case and highlight military relevant issues that include efficient diagnostic algorithms in the field and transfusion of scarce blood products. PMID- 12901475 TI - Fewer students select family medicine. PMID- 12901476 TI - More critical analysis needed. PMID- 12901477 TI - Show us the numbers. PMID- 12901478 TI - Is expensive medicine worth it? PMID- 12901479 TI - Reflecting on a week just past. PMID- 12901480 TI - Ionizing radiation during pregnancy. AB - QUESTION: One of my patients had a computed tomography scan of her abdomen a week ago and has just found out she is 7 weeks pregnant. What should I tell her about the pregnancy and the risk to her fetus? ANSWER: Your patient is not at increased risk of miscarriage or major congenital fetal malformations due to radiation exposure. Her risk is similar to that of the general population (ie, 1% to 3%). PMID- 12901481 TI - Reporting drug seekers. PMID- 12901482 TI - Managing opioid dependence. Comparing buprenorphine with methadone. PMID- 12901483 TI - Thiazides first-line treatment for hypertension. PMID- 12901484 TI - Oncologists and family physicians. Using a standardized letter to improve communication. AB - PROBLEM BEING ADDRESSED: Communication between oncologists at a regional cancer centre and family physicians caring for palliative cancer patients in the community was ineffective. OBJECTIVE OF PROGRAM: To improve communication between oncologists and family physicians by routine use of a template for dictated letters concerning follow-up care. PROGRAM DESCRIPTION: A consultation letter template was constructed and tested at a single clinic. The template was designed to guide oncologists dictating letters to family physicians for patients' follow up visits. Effectiveness of the standardized letter was evaluated with a before after survey. CONCLUSION: Using the template letter improved communication with respect to the relevance, timeliness, format, and amount of information. As care for patients at the end of life increasingly shifts to the community, ongoing efforts are required to improve communication between cancer centres and primary care physicians. PMID- 12901485 TI - Case report: Molluscum contagiosum. Toxic shock syndrome following cantharidin treatment. PMID- 12901486 TI - Cancer follow-up care. Patients' perspectives. AB - OBJECTIVE: To assess family physicians' and specialists' involvement in cancer follow-up care and how this involvement is perceived by cancer patients. DESIGN: Self-administered survey. SETTING: A health region in New Brunswick. PARTICIPANTS: A nonprobability cluster sample of 183 participants. MAIN OUTCOME MEASURES: Patients' perceptions of cancer follow-up care. RESULTS: More than a third of participants (36%) were not sure which physician was in charge of their cancer follow-up care. As part of follow-up care, 80% of participants wanted counseling from their family physicians, but only 20% received it. About a third of participants (32%) were not satisfied with the follow-up care provided by their family physicians. In contrast, only 18% of participants were dissatisfied with the follow-up care provided by specialists. Older participants were more satisfied with cancer follow-up care than younger participants. CONCLUSION: Cancer follow-up care is increasingly becoming part of family physicians' practices. Family physicians need to develop an approach that addresses patients' needs, particularly in the area of emotional support. PMID- 12901487 TI - Knowledge and attitudes regarding care of leg ulcers. Survey of family physicians. AB - OBJECTIVE: To determine family physicians' perceptions of and attitudes toward leg ulcer care and awareness of effective treatments for venous leg ulcers. DESIGN: Self-administered, cross-sectional faxed and mailed survey. SETTING: Ottawa-Carleton, Ont. PARTICIPANTS: All physicians in the region who were members of the College of Family Physicians of Canada. RESULTS: Response rate was 62%. During 1 month, 107 physicians reported having 226 patients with leg ulcers; only a few patients had had ultrasound assessment. Few physicians (16%) were confident about managing leg ulcers; 61% reported not knowing enough about wound-care products. More than 50% were unaware that compression is effective treatment for venous ulcers. Problems reported were lack of evidence-based clinical practice guidelines for leg ulcer care (82%); absence of evidence-based protocols in home care agencies (72%); lack of access to wound-care products (69%) and wound-care centres (66%); and poor communication among health care workers (60%). CONCLUSION: Better access to diagnostic assessments and use of compression therapy for venous leg ulcers would improve care. PMID- 12901489 TI - Care today and residents' care in the future. PMID- 12901488 TI - Colorectal cancer screening: in your office now? PMID- 12901490 TI - Making a case for transdisciplinarity. PMID- 12901491 TI - Radiation-induced progressive decreasing in the expression of reverse transcriptase gene of hEST2 and telomerase activity. AB - OBJECTIVES: In order to identify the relationship between telomerase and the biological effect of radiation injury, and investigate the role of human telomerase catalytic subunit gene (hEST2) reverse transcriptase(RT) segment in the expression of telomerase activity. METHODS: Tumor HeLa cells, KB cells and A431 cells were employed to measure the change in telomerase activity after 60Co ray irradiation at RNA level and protein level. Quantitative PCR and Northern blotting were used to determine the expression of hEST2 RT segment that encodes seven motifs of the human telomeres, a PCR-based telomeric repeat amplification protocol (TRAP) was used to assay telomerase activity after exposure to radiation. RESULTS: Both of telomerase activity and the expression hEST2 RT segment were decreased with increasing dosage of radiation. In addition, testing the expression of motifs domain is similar to the measurement of telomerase activity. CONCLUSION: The detection of the hEST2 RT segment by Northern blotting and quantitative PCR are new methods for testing telomerase activity. Furthermore, radiation can cause a dose-dependent decrease in telomerase activity. The effect of radiation on telomerase is one possible reason for the death of cancer cells after irradiation. PMID- 12901492 TI - A preliminary study on comparison and fusion of metabolic images of PET with anatomic images of CT and MRI. AB - OBJECTIVE: To compare and match metabolic images of PET with anatomic images of CT and MRI. METHODS: The CT or MRI images of the patients were obtained through a photo scanner, andthen transferred to the remote workstation of PET scanner with a floppy disk. A fusion method was developed to match the 2-dimensional CT or MRI slices with the correlative slices of 3-dimensional volume PET images. RESULTS: Twenty-nine metabolically changed foci were accurately localized in 21 epilepsy patients' MRI images, while MRI alone had only 6 true positive findings. In 53 cancer or suspicious cancer patients, 53 positive lesions detected by PET were compared and matched with the corresponding lesions in CT or MRI images, in which 10 lesions were missed. On the other hand, 23 lesions detected from the patients' CT or MRI images were negative or with low uptake in the PET images, and they were finally proved as benign. CONCLUSIONS: Comparing and matching metabolic images with anatomic images helped obtain a full understanding about the lesion and its peripheral structures. The fusion method was simple, practical and useful for localizing metabolically changed lesions. PMID- 12901494 TI - IgE production is involved in butyrate-enhanced NK cell activity in vivo. AB - It has been demonstrated that patients with asthma have a large number of NK cells and show a stronger NK activity. These results indicate that NK cell activity may be related to total IgE level in serum in healthy subjects. Previously, we have found that sodium butyrate (NaBu) markedly enhanced the IL-4 induced IgE production in the LPS-stimulated murine splenocytes in vitro, and inductive rat IgE production in vivo, and enhanced the NK cell activity ex vivo. We hypothesized that the IgE production might be involved in butyrate-enhanced NK cell activity in vivo. Mice were intraperitoneally treated/immunized with NaBu or/and Ascaris suum extract (ASC), and the spleen NK cell activity was evaluated. Furthermore, the effect of serum (NAS) on IL-2- or IFN-gamma-induced spleen NK cell activity was determined. The spleen NK cell activity and IL-2- or IFN-gamma induced spleen NK cell activity of mice treated/immunized with NaBu or/and ASC were stronger than those of untreated/unimmunized mice. Although IL-4 blocked IL 2 (100 U/ml)- or IFN-gamma (100 U/ml)-induced increase in NK cell activity, these NK cell activities in mice treated/immunized with NaBu/ASC were not inhibited. IgE production showed a tendency to rise in NaBu-treated mice serum, and a synergistic effect was observed with treatment of NaBu and ASC. Moreover, the NAS significantly increased IL-2(25 U/ml)-or IFN-gamma(25 U/ml)-induced NK cell activity, and its effect was inhibited by anti-mouse IgE mAb. These data show that IgE plays an important role in NAS-enhanced IL-2/IFN-gamma-induced NK cell activity, and IL-4 does not inhibit IgE and IL-2/IFN-gamma-induced NK cell activity in mice. PMID- 12901493 TI - Association between low-density lipoprotein receptor-related protein gene, butyrylcholinesterase gene and Alzheimer' s disease in Chinese. AB - OBJECTIVE: To research the relations between low-density lipoprotein receptor related protein gene (LRP) polymorphism, butyrylcholinesterase gene (BchE) polymorphism and Alzheimer's disease (AD) in Chinese. METHODS: The gene polymorphisms of LRP and BchE were genotyped in 38 AD cases and 40 controls with polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) methods. AD groups were classified according to the LRP C/C genotype and compared with matched controls. RESULTS: AD group had higher frequencies of C/C homozygote (81.69% vs 60.0%, P < 0.05) and of C allele (89.5% vs 76.3%, P < 0.05), with no significant difference between any of these LRP genotypes classified AD groups and their respective control groups. CONCLUSIONS: A positive correlation was found between LRP gene polymorphism and AD, but not between BchE gene polymorphism and AD in Chinese AD cases. PMID- 12901496 TI - Intracellular redistribution of cardiac endothelin-1 receptor in rat during myocardial hypertrophy. AB - OBJECTIVE: In a model of rat cardiac hypertrophy, the changes in the distribution of ET-1 receptors in two subcellular fractions, the sarcolemma and the light vesicles during myocardial hypertrophy were studied. METHODS: Cardiac hypertrophy was produced by placing a constricting clip around the suprarenal abdominal aorta of rats, and ET-1 receptor was assayed with radioactive analysis method. RESULTS: It was found that plasma and ventricular ET-1 levels increased significantly on week 2 and week 4 of pressure overload. ET-1 binding studies showed that during myocardial hypertrophy, the maximum binding capacity (B(max)) was increased by 41% (P < 0.01) and 65% (P < 0.01) in sarcolemma in H-2 week and H-4 week groups, but was decreased by 24% (P < 0.01) and 21% (P < 0.01) in light vesicles. The sum of B(max) of sarcolemmal and light vesicle fractions was increased by 33% (P < 0.01) and 57% (P < 0.01) in group H-2 week and H-4 week, respectively. CONCLUSION: ET-1 receptors in rat heart were externalized from light vesicles to sarcolemma, which may contribute to the development of myocardial hypertrophy. PMID- 12901495 TI - Effect of angiotensin II receptor antagonist and endothelin receptor antagonist on nitroglycerin tolerance in rats. AB - OBJECTIVE: To investigate whether angiotensin II receptor antagonist and endothelin receptor antagonist can improve the nitroglycerin (Nit) tolerance in vivo. METHODS: Twenty-four rats were divided into 4 groups (n = 6, each): Control group, Nitroglycerin (Nit) group, Nit + bosentan group and Nit + losartan group. Nitroglycerin tolerance was induced by 2-day treatment of (10 mg x kg(-1) nitroglycerin patch(0.05 mg/h). Angiotensin receptor antagonist losartan(10 mg x kg(-1) x d(-1)) and endothelin receptor antagonist bosentan (100 mg x kg(-1) x d( 1)) were given by gavage for 2 days respectively. RESULTS: The least hypotensive response to sodium nitroprusside (SNP) was observed in Nit group. The effective percentages of hypotensive response to SNP were increased in both Nit + losartan group and Nit + bosentan group compared with Nit group [(31.95 +/- 4.45) % vs (21.00 +/- 3.69) %, P < 0.01 and (33.18 +/- 6.16) % vs (21.00 +/- 3.69) %, P < 0.01, respectively]. The maximal vessel relaxation induced by SNP was the same in 4 different groups but the highest EC50 (concentration which produces 50% of the maximal response to SNP) was found in tolerant group[(34 +/- 10) nmol/L, P < 0.01]. The ET-1 amounts in plasma and vascular tissue were markedly increased by 54% and 60% in Nit group compared with those in control group( P < 0.01). The ET 1 amounts in plasma and vascular tissue were decreased by 30% and 37% in Nit + losartan group compared with those in Nit group (P < 0.01). CONCLUSION: Endothelin receptor antagonist and angiotensin II receptor antagonist could prevent against the Nit tolerance. PMID- 12901497 TI - The distraction reduction fixation system and its application in spondylolisthesis. AB - OBJECTIVE: An internal fixation apparatus--distraction reduction fixation system (DRFS) was designed to satisfy the clinical needs for spondylolisthesis. METHODS: Since 1996, 53 patients were treated with DRFS. Among them, 35 had spondylolisthesis, 12 had lumbar canal stenosis accompanied with instability, 2 had vertebral tumors and 4 suffered from spinal fracture. The average age was 53.6 years old (ranged 24 approximately 72 yrs). The mean time for follow-up was 30.6 months (16 approximately 44 months). RESULTS: The slip rate was 0.15 +/- 0.10 before operation, and decreased to 0.09 +/- 0.07 after operation. Entire slip reposition was achieved in 19 cases (54.3%). The change in height of the intervertebral space within the fixation segments was 0.7 +/- 0.17. CONCLUSION: DRFS achieved better results for spondylolisthesis less II degree and no other adverse effects were found. Compared with other foreign and domestic techniques, it had advantages in less implants, less operation gears required and ease to utilize in operation. It was proved to be an ideal internal fixation apparatus. PMID- 12901498 TI - Three-dimensional correction of scoliosis using TSRH instrumentation. AB - OBJECTIVE: To evaluate the results of TSRH instrumentation in the correction of coronal, sagittal and rotational deformity of scoliosis. METHODS: From January 1998 to December 1999, thirty-two consecutive patients (6 males, 26 females)with scoliosis underwent anterior or posterior spinal instrumentation and fusion using TSRH instrumentation. Of these cases, 21 were idiopathic scoliosis and 11 were congenital scoliosis. The average age at surgery was 16.4 years (range, 11 approximately 45 years). The mean Cobb angle at surgery was 71.2 degrees range, 44 degrees approximately 125 degrees) in the coronal plane, and 49. degrees range, 16 degrees aprroximately 67degrees in the sagittal plane. Rotational deformity (Nash-Moe) ranged from I to III degree. Preoperative apical translation averaged 4.8 cm (range, 3 approximately 9 cm). RESULTS: The average follow-up duration was 13.3 months (range, 10 approximately 24 months). At the final follow up, the mean Cobb angle in the coronal plane was 26.6 (range, 10 degrees approximately 73 degrees), with a 63.8% of improvement. Sagittal alignment was well maintained with a mean Cobb angle of 28 degrees (range, 10 degrees approximatelky 45 degrees). The average correction of rotation of the apical vertebra was I degree. The average apical translation was 1.6 cm (range, 0.5 approximately 5.0 cm) representing a correction rate of 66, 7%. Complication was noted in two cases with an incidence of 3.1%, one case had superficial infection and the other one had lower hook dislocation. There was no neurologic deficit and pseudoarthrodesis in this series. CONCLUSION: TSRH instrumentation is an effective and convenient three-dimensional correction system with a lower rate of complication, which can not only correct the coronal and rotational deformity, but maintain the sagittal alignment as well. PMID- 12901499 TI - Cartilage-like phenomenon in the anterior cruciate ligamenta. AB - OBJECTIVE: To detect histological characteristic of anterior cruciate ligament (ACL) and medial collateral ligament (MCL). METHODS: In each of 20 skeletally mature male mongrels and 4 men, the ACL and MCL were examined by standard hematoxylin-eosin procedure and toluidine blue staining for histologic observation. RESULTS: The fibroblasts in medial collateral are elongated to spindle shape and aligned in a row between the bundles of collagenous fibers. Toluidine blue staining is negative. The anterior cruciate ligament demonstrated more heterogenous cell types and arrangement. It had three major cell forms: spindle, round and ovoid type, which were shorter but greater than the cells in medial collateral ligament. Toluidine blue staining was positive in anterior cruciate ligament. Most cells in anterior cruciate ligament were enclosed within lacunae. CONCLUSION: This study suggests that the ACL has different histological characteristics from MCL, and is more cartilage-like in nature. PMID- 12901500 TI - Magnetic resonance imaging of acute central cord syndrome: correlation with prognosis. AB - OBJECTIVE: To report the magnetic resonance imaging (MR) findings of acute cervical central cord syndrome and to determine their correlation with the prognosis. METHODS: MRI findings of 35 patients with acute central cord syndrome were studied and compared with the recovery rate of ASIA score at presentation and in follow-up. RESULTS: MRI data demonstrated spinal cord compression for 32 patients, spinal cord swelling for 16 patients, and abnormal signal intensity within the spinal cord for 19 patients, including 14 with edema and 3 with hematoma. No significant difference of the recovery rate was noted between the patients treated nonoperatively and operatively (P > 0.05). There was a significant inverse correlation between the recovery rate and the degree of spinal cord compression as shown in MRI scans (P < 0.01). The presence of hematoma in MRI scans was associated with poor prognosis, as demonstrated by a significant difference of the recovery rate (P < 0.01) among the patients with normal intensity, edema and hematoma within the spinal cord. CONCLUSIONS: MRI scans provide an efficient assistance for decision-making and accurate prognostic information regarding neurological function, and therefore should routinely be performed within the early phase of acute central cord syndrome. PMID- 12901501 TI - Laparoscopic myomectomy. AB - OBJECTIVE: To investigate the indications, surgical techniques and complications of laparoscopic myomectomy. MATERIALS AND METHODS: A retrospective study was carried out in 74 patients with fibroids > 3 cm from March, 1995 until May, 2000 at PUMC Hospital. Indications for surgery were symptomatic fibroids (20 cases), mainly pain or urine frequency; progressively increasing fibroid size (7 cases); coexistent adnexal pathology(26 cases) and infertility(21 cases). RESULTS: The number of fibroids of each patient varied from 1 to 4 with single fibroid of 62 cases (83.7%). The fibroids were located in anterior wall (30 cases), posterior wall (23 cases) and fundus (21 cases). A total of 93 fibroids were removed from these patients including 16 intramural fibroids and 77 subserous fibroids. The size of dominant fibroids ranged from 3 approximately 8 cm (mean 4.8 cm). In 19 cases (25.6%), the uterine wall was sutured in one layer. Mean duration of operation was 73 minutes and mean blood loss was 82 ml. Longer operating time and more blood loss were observed in patients with fibroids > or = 4 cm than those with fibroids < 4 cm. The difference was statistically significant ( P < 0.05). Mean postoperative hospital stay was 3.2 days and overall complication rate was 1.4%. The average postoperative follow-up period was 22 months (1 approximately 62 months). All the patients with symptoms showed remission of their complaints at 2-month follow-up. Recurrence of fibroid occurred in 1 case 1 year after initial operation and second laparoscopic myomectomy was given to her successfully. Five patients became pregnant. The pregnancy was uneventful and proceeded to selective caesarean section at term pregnancy in 4 cases. One miscarriage occured at 8 weeks in the 5th case. No adhesions at myomectomy site were found in these 5 patients. CONCLUSIONS: Our study suggests the feasibility of laparoscopic myomectomy in selected patients, which leads to effectiveness, low complication rate and satisfactory remission of symptoms. Further study on recurrence and fertility must be continued. PMID- 12901502 TI - The mechanism of cerebral evoked potentials by repetitive magnetic stimulation of gastrocnemius muscle in Duchenne muscular dystrophy. AB - OBJECTIVE: To study the features and mechanism of the cerebral evoked potentials by repetitive stimulation of calf muscle in Duchenne muscular dystrophy (DMD) patients with obvious muscular dystrophy and psuedohypertrophy. METHODS: Cerebral evoked potentials by stimulation of calf muscles and somatosensory evoked potentials (SEPs) by the stimulation of posterior tibial nerves at ankle were measured in 10 patients with DMD and 10 normal controls matched with gender and age. The intensity of the magnetic stimulation was at 30% of maximal output (2.1 Tesla, MagPro magnetic stimulator, Dantec) and the frequency was 1 Hz. The low intensity of magnetic stimulation was just sufficient to produce a contraction of the muscle belly underneath the coil. Recording electrode was placed at 2 cm posterior to the Cz, reference to Fpz. The latencies of N33, P38, N48 and P55 and amplitude (P38 - N48) were recorded. SEPs were recorded by routine methods. RESULTS: In normal subjects, the amplitudes of cerebral evoked potentials by magnetic stimulation of calf muscle was 40% lower than that by electrical stimulation of the posterior tibial nerves at ankle. The latency of P38 was 2.9 +/- 2.1 ms longer compared with electrical stimulation of the posterior tibial nerves at ankle. In 6 patients, P38 latency from magnetic stimulation was remarkably prolonged (P < 0.01), and in 4 patients, there was no remarkable response. SEPs evoked by electrical stimulation were normal in all of the patients. CONCLUSION: DMD is an available model for the study of mechanism of cerebral evoked potentials by magnetic stimulating muscle. We can conclude that the responses from magnetic stimulation were produced by muscle input. The abnormal responses in patients may relate to decreased input of muscle by stimulating dystrophic and psedohypertrophic muscle. PMID- 12901503 TI - Preliminary studies on associations of IDDM3, IDDM4, IDDM5 and IDDM8 with IDDM in Chengdu population. AB - OBJECTIVE: To study the associations of IDDM3, IDDM4, IDDM5 and IDDM8 with insulin-dependent diabetes mellitus (IDDM). METHODS: The polymorphisms of short tandem repeat (STR) loci D15S657, D11S1369, D6S2420 and D6S503, linked to IDDM3, IDDM4, IDDM5 and IDDM8 respectively, were studied by polymerase chain reaction and polyacrylamide gel electrophoresis (PCR-PAGE) followed by direct sequencing of PCR products in 105 normal Chinese Han nationality subjects and 48 patients with IDDM. RESULTS: The allele frequencies of allele A5 at D15S657 locus, allele A5 at D11S1369 locus and allele A4 at D6S2420 locus were increased significantly in patients with IDDM compared to those in the control group. No difference in the allele frequencies at D6S503 locus was observed between IDDM and control group. CONCLUSION: IDDM3, IDDM4, IDDM5 but not IDDM8 may be associated with IDDM in Chinese Han nationality population. PMID- 12901504 TI - A case report of dopa-responsive dystonia. PMID- 12901505 TI - Differentiation and malignant suppression induced by mouse erythroid differentiation and denucleation factor on mouse erythroleukemia cells. AB - OBJECTIVE: To investigate the roles of mouse erythroid differentiation and denucleation factor (MEDDF), a novel factor cloned in our laboratory recently, in erythroid terminal differentiation. METHODS: Mouse erythroleukemia (MEL) cells were transfected with eukaryotic expression plasmid pcDNA-MEDDF. Then we investigated the changes on characteristics of cell growth by analyzing cells growth rate, mitotic index and colony-forming rate in semi-solid medium. The expressions of c-myc and beta-globin genes were analysed by semi-quantitative RT PCR. RESULTS: MEL cells transfected with pcDNA-MEDDF showed significant lower growth rate, mitotic index, and colony-forming rate in semi-solid medium (P<0.01). The percentage of benzidine-positive cells was 32.8% after transfection. The expression of beta-globin in cells transfected with pcDNA-MEDDF was 3.43 times higher than that of control (MEL transfected with blank vector, pcDNA3.1), and the expression of c-myc decreased by 66.3%. CONCLUSIONS: MEDDF can induce differentiation of MEL cell and suppress its malignancy. PMID- 12901507 TI - Effects of alendronate on structural properties of trabecular bone in dogs. AB - OBJECTIVE: To evaluate the effects of alendronate on the structural properties of trabecular bone. METHODS: Alendronate was administered at a daily p.o. dose of 0.5 mg/kg over a 12-week period in hound dogs (n = 8 for both the control and treated group), and the structural indices of the lumbar vertebral (L1 and L2) trabecular bone were assessed directly from 3-D images. RESULTS: Treatment with alendronate increased bone volume fraction by 9.5% and 7.7% in L1 and L2 respectively. Trabecular thickness significantly increased after alendronate treatment, whereas trabecular separation remained constant. The degree of anisotropy for the alendronate-treated group was decreased compared with that of the control group. Bone surface to volume ratio declined significantly in the alendronate-treated group, whereas alendronate induced a higher bone surface density. CONCLUSION: Alendronate increased the structural properties of canine trabecular bone after short-term treatment at a dose of 0.5 mg x k(-1) x day(-1). PMID- 12901506 TI - The role of IGF-1 gene expression abnormality in pathogenesis of diabetic peripheral neuropathy. AB - OBJECTIVE: To explore the role of insulin-like growth factor 1 (IGF-1) gene expression abnormality in neurotrophic causes of diabetic peripheral neurophathy. METHODS: Diabetes was induced in Sprague Dawley rats by alloxan. The parameters were measured as follows: IGF-1 mRNA by revere transcriptase-polymer chain reaction (RT-PCR); IGF-1 peptide by enzyme-linked immunosorbent assay (ELISA); electrophysiological parameters of nerves by evoked electromyogram; morphometric evaluation of sciatic nerves under light microscope and transmission electron microscope. RESULTS: During early diabetic stage, IGF-1 mRNA [(0.430+/-0.031) vs. (0.370+/-0.016), P<0.01, (0.430+/-0.031) vs. (0.280+/-0.010), P<0.001, respectively], IGF-1 peptide contents [(38.44+/-3.60) ng/mg vs. (30.06+/-2.41) ng/mg, P<0.01, (38.44+/-3.6) ng/mg vs. (3.71+/-2.70) ng/mg P<0.001, respectively] in sciatic nerve tissue reduced in diabetic rats with hyperglycemia and varied with severity of state when compared with non-diabetic control rats, and further gradually down-regulated in the diabetic rats with duration of diabetes [IGF-1 mRNA (0.320+/-0.021) to approximately (0.230+/-0.060); IGF-1 peptide (28.80+/ 3.30) to approximately (19.51+/-1.80) ng/mg]. Furthermore, they correlated with nerve functional (sensory nerve conduction velocity: r=0.741, P<0.001; amplitude of evoked potential: r=0.716, P<0.001, respectively) and structural abnormality (axonal area r=0.81, P<0.001) of sciatic nerve. No difference was found in the above parameters between diabetic rats with euglycemia and non-diabetic control group. CONCLUSION: IGF-1 gene expression in tissues was down-regulated from early diabetic stage, and varied with the severity and duration of diabetic state. The decrement in IGF-1 level might contribute to the initiation and development of diabetic neuropathy via autocrine or paracrine pathway. PMID- 12901508 TI - Production of phage-displayed anti-idiotypic antibody single chain variable fragments to MG7 monoclonal antibody directed against gastric carcinoma. AB - OBJECTIVE: To generate phage-displayed anti-idiotypic antibody single chain variable fragments (anti-Id ScFv) to MG7 monoclonal antibody (McAb) directed against gastric carcinoma so as to lay a foundation for developing anti-Id ScFv vaccine of the cancer. METHODS: Balb/c mice were immunized i.p. with MG7 McAb conjugated with keyhole limpet hemocyanin (KLH), and mRNA was isolated from the spleens of the immunized mice. Heavy and light chain (VH and VL) genes of antibody were amplified separately and assembled into ScFv genes with a linker DNA by PCR. The ScFv genes were ligated into the phagemid vector pCANTAB5E and the ligated sample was transformed into competent E. coli TG1. The transformants were infected with M13K07 helper phage to yield recombinant phages displaying ScFv on the tips of M13 phage. After 4 rounds of panning with MG7, the MG7 positive clones were selected by ELISA from the enriched phages. The types of the anti-Id ScFv displayed on the selected phage clones were preliminarily identified by competition ELISA. RESULTS: The VH, VL and ScFv DNAs were about 340 bp, 320 bp and 750 bp respectively. Twenty-four MG7-positive clones were selected from 60 enriched phage clones, among which 5 displayed beta or gamma type anti-Id ScFv. CONCLUSION: The anti-Id ScFv to MG7 McAb can be successfully selected by recombinant phage antibody technique, which paves a way for the study of prevention and cure of gastric carcinoma by using anti-Id ScFv. PMID- 12901509 TI - Nanoparticle as a new gene transferring vector in specific expression gene. AB - OBJECTIVE: To evaluate the possibility and efficiency of nanoparticle as a new vector in specific gene transference. METHODS: Nanoparticle-DNA complex was prepared with Poly-dl-lactic-co-glycolic acid (PLGA) bearing anti-sense monocyte chemotactic protein-1 (A-MCP-1), a specific expression gene, and the package efficiency, release progress in vitro, and the size of the complex were determined. The possibility of the new vector was evaluated with genomic DNA PCR by transferring gene into cultured smooth muscle cells (SMC), cationic lipids as a control. For study in vivo, jugular vein-to-artery bypass grafting procedures were performed on 20 New Zealand white rabbits, of which 6 grafts were transferred with nanoparticle-A-MCP-1 (200 microg), 6 with A-MCP-1 (200 microg) by cationic liposome, 4 with LNCX plasmid, and 4 as control. Fourteen days after the grafts were harvested, the expression of A-MCP-1 and its effect on MCP-1 in vein grafts were detected by dot blot, and the morphologic evaluation of grafts was performed. RESULTS: The package efficiency of the nanoparticle-DNA complex was 0.9%, release progress in vitro lasted 2 weeks, and the size ranged from 150 to 300 nm. SMC genomic DNA PCR showed that A-MCP-1 gene could be successfully transfected into cells by nanoparticle. The study in vivo indicated that A-MCP-1 mRNA was expressed in both local gene delivery groups, nanoparticle and liposome, meanwhile, MCP-1 expression in vein grafts was significantly inhibited and neointimal hyperplasia was notably reduced. CONCLUSION: Nanoparticle can act as a vector to transfect specific gene. PMID- 12901510 TI - Screening for mutations in a novel retinal-specific gene among Chinese patients with retinitis pigmentosa. AB - OBJECTIVE: To identify and evaluate mutations in the RP1 gene among Chinese patients with retinitis pigmentosa (RP). METHODS: Leukocyte DNA of 92 RP patients were collected in Hong Kong. Sequence changes of the entire coding region of the RP1 gene were examined using PCR, conformation sensitive gel electrophoresis and DNA sequencing. RESULTS: In total, 1 nonsense mutation and 1 nonsense variant as well as 10 missense alterations were identified in the RP1 gene, among which, Arg677Ter was found in 1 RP patient and another nonsense variant, Arg1933Ter, was identified in 3 normal individuals and 1 patient with Stargardt's disease, suggesting its nonpathogenicity. Arg77Ter is expected to lead to large disruptions of the encoded protein. CONCLUSIONS: The nonpathogenicity of Arg1933Ter indicates that the C-terminal 224 residues of RP1 protein may be not critical for RP1. The most C-terminal truncation previously reported was due to Tyr1053 (1-bp del) and occurred in RP patients. Thus RP can be caused by reduction in the level of the region of RP1 protein after codon 1052 but before 1933. To ascertain such a proposition, genotypes of more RP patients may reveal more RP causative mutations and more sequence alterations different than those of other ethnic groups. PMID- 12901511 TI - Association of plasma homocysteine level and N5,N10-methylenetetrahydrofolate reductase gene polymorphism with cerebral infarction. AB - OBJECTIVE: To investigate the relationship of plasma homocysteine (Hcy) level to stroke and genetic factor to elevated plasma Hcy level. METHODS: The plasma Hcy level was measured by capillary electrophoresis-ultraviolet detection and the gene polymorphism of N5,N10-methylenetetrahydrofolate reductase (MTHFR) was studied with PCR-RFLP assay in 43 patients with cortical cerebral infarction and 42 healthy control. RESULTS: The plasma Hcy level of the patients (19.3 +/- 6.0 micromol/L) was markedly higher than that of the controls (13.7 +/- 5.4 micromol/L) (t = 4.16, P < 0.001). There are 3 genotypes, C/C, C/T and T/T, about base-variation of MTHFR gene at locus 677. The plasma Hcy level of the subjects with T/T genotype was higher than that of subjects with other genotypes. However, the frequencies of each genotype and allele were not significantly different between the patients and the controls. CONCLUSIONS: The elevated plasma Hcy level is a risk factor for atherothrombotic cerebral infarction, and is related to the C-->T mutation at locus 677 of MTHFR gene. PMID- 12901513 TI - Pregnancy and its outcome in women with malformed uterus. AB - OBJECTIVE: To analyze the clinical characteristics of fertility and pregnancy in women with congenital uterine malformations and explore optimal treatments to improve the prognosis. METHODS: A retrospective study was conducted on the fertility and obstetric outcome in 153 patients with uterine malformations treated in our hospital from January 1984 to December 1998. Twenty-seven cases with other kinds of genital and/or urinary anomalies but with normal uterus during the same period were enrolled as the control group. RESULTS: The infertility rate was 26.6% (34/128), the miscarriage rate 44.3% (86/194), premature birth rate 9.3% (18/194), abnormal fetal presentation rate 28.4% (29/102), the cesarean section rate 61.8% (63/102), and the perinatal mortality rate 11.8% (12/102). CONCLUSION: Women with congenital uterine malformation usually have higher incidence of infertility and complications during pregnancy and delivery. Bicornuate and septate uterus can be associated with poor obstetric outcome. PMID- 12901512 TI - Comparison of the effects of losartan, enalapril and their combination in the prevention of left ventricular remodeling after acute myocardial infarction in the rat. AB - OBJECTIVES: To compare the effects of losartan, enalapril and their combination in the prevention of left ventricular remodeling (LVRM) after acute myocardial infarction (AMI) in the rat. METHODS: AMI model was induced in female SD rats by ligating left coronary artery. Forty-eight hours after the procedure, 83 surviving rats were randomized into one of the following 4 groups : 1) AMI control group (n = 19), 2) losartan group (n = 22, 3 mg x kg(-1) x d(-1)), 3) enalapril group (n = 20, 1 mg x kg(-1) x d(-1)), 4) losartan-enalapril combinative group (n = 22, 3 and 1 mg x kg(-1) x d(-1) respectively). 5) Sham operated group (n = 10) and 6) normal rats group (n = 10) were selected randomly to serve as non-infarction controls. Losartan and enalapril were delivered by direct gastric gavage. After 4 weeks of medical therapy, hemodynamic studies were performed in each group, then the rat hearts were fixed with 10% formalin and pathologic analysis on them was performed. Complete experimental data was obtained in 56 rats, comprising 1) AMI controls (n = 11), 2) losartan group (n = 10), 3) enalapril group (n = 10), 4) the combination of losartan and enalapril group (n = 11), 5) sham-operated group (n = 6) and 6) normal controls (n = 8). RESULTS: There were no significant differences among the 4 AMI groups in MI size (41.7% to approximately 43.4%, all P > 0.05). Compared with sham group, the left ventricular (LV) end diastolic pressure (LVEDP), volume (LVV), long and short axis length (L and D), as well as LV absolute and relative weight (LVAW and LVRW) in AMI group were all significantly increased (P < 0.05 to approximately 0.001); whereas the maximum left ventricular pressure rising and dropping rates (+/- dp/dt) and their corrected values by LV systolic pressure (+/- dp/dt/LVSP) were significantly reduced (all P < 0.001), indicating LVRM occurred and LV systolic and diastolic function impaired after AMI. Compared with AMI group, LVEDP, LVV, LVAW and LVRW were all significantly decreased (P < 0.05 to approximately 0.001); while +/- dp/dt/LVSP were significantly enhanced in all 3 treatment groups (P < 0.05 to approximately 0.001) except -dp/dt/LVSP in losartan group (P > 0.05). There were no significant differences in the above indices among the 3 treatment groups (all P > 0.05). CONCLUSION: Both losartan and enalapril can prevent from LVRM after AMI in the rat and improve LV function with equivalent effects. There seems no additive effect when the 2 drugs are used in combination. PMID- 12901514 TI - Pulmonary embolism during pregnancy and the postpartum period: report of 2 cases. AB - OBJECTIVE: To investigate the high risk factors, clinical features, diagnosis and treatment of pulmonary embolism (PE) during pregnancy and the postpartum period. METHODS: Two cases of PE during pregnancy and postpartum period were reported retrospectively. RESULTS: The first case was a pregnant woman with congenital heart disease at 39-week gestation. She underwent a successful cesarean section (CS) because of heart disease, but she manifested tachycardia, tachypnea, cyanosis, and dyspnea suddenly on the 10th day after CS and died soon after the onset of these symptoms. The diagnosis of PE was highly suspected clinically. The second case was a twin-pregnancy woman at 35-week gestation. Because of severe perinatal myocardiopathy, cesarean section was performed. Unfortunately, the patient died abruptly on the operation table. The diagnosis of PE was confirmed by autopsy. CONCLUSIONS: The maternal mortality of PE during pregnancy and postpartum period is quite high. Whenever there is any doubt, objective examinations for PE should be started early in order to strive for the chances of the anticoagulant therapy. PMID- 12901515 TI - Heat shock protein gp96 and cancer immunotherapy. AB - Heat shock protein gp96 is a highly conserved and monomorphic glycoprotein in the endoplasmic reticulum. It functions as molecular chaperone and can associate with a variety of antigenic peptides noncovalently in vivo and in vitro. Recent studies have indicated that gp96 molecules participate in major histocompatibility complex class I-restricted antigen presentation pathway. Immunization of mice with gp96 preparations isolated from cancer cells can elicit a cancer-specific protective T cell immune response that is recallable, which is a prerequisite for gp96 as a therapeutic vaccine against cancers. The immunogenicity of gp96 molecules has been attributed to the antigenic peptides associated with them. These phenomena provide a new pathway for cancer immunotherapy. The mechanism that the gp96-peptide complex induces specific immune response and the explorations for gp96-peptide complex as a therapeutic cancer vaccine are reviewed. PMID- 12901516 TI - Effects of hCD80-adenovirus on B16-F10 cells. PMID- 12901517 TI - Impairment of cardiac function by acute carbon monoxide poisoning. PMID- 12901519 TI - Effect of nimodipine, D-AP-5 and mild hypothermia on cultured neuronal [H+]i following fluid percussing injury. PMID- 12901518 TI - Serum levels of soluble Fas ligand and soluble Fas receptor in patients with chronic congestive heart failure. PMID- 12901521 TI - The influence of apolipoprotein B and E gene polymorphisms on the response to simvastatin therapy in patients with hyperlipidemia. AB - OBJECTIVE: To investigate the effect of apolipoprotein B (apoB) and E (apoE) genetic variations on lipid profile at baseline (before treatment), and also on the subsequent response to simvastatin therapy. METHODS: Eighty-eight patients with hyperlipidemia were treated with simvastatin 5mg daily. The plasma levels of total cholesterol (TC) and low-density lipoprotein cholesterol (LDL-C), triglyceride (TG) and apo B were measured pre-treatment and at the end of the 4th, 8th and 12th post-treatment week. Polymorphisms of apoB at XbaI locus and apoE were determined by restriction fragment length polymorphism (RFLP). RESULTS: In all patients, relative frequencies of X- allele and X+ allele were 0.943 and 0.057 for apoB gene respectively. For apoE gene the relative frequency of epsilon2 allele was determined as 0.182, epsilon3 as 0.580 and epsilon4 as 0.238. The reduction in TC level was more prominent in patients carrying X- allele than in those with X+ allele following treatment (-23. 9% vs. -13. 6%, P < 0. 05). Compared with patients carrying epsilon3 or epsilon4 allele, those with epsilon2 allele showed a significantly higher percentage in reduction of apoB level after treatment (P < 0.05). CONCLUSION: The relative frequency of apoB X+ allele is high in patients with hyperlipidemia, in whom the TC-lowering efficacy is decreased following treatment of simvastatin. The relative frequencies of epsilon2 and epsilon4 are also high in hyperlipidemic patients, and the epsilon2 allele is associated with reduction in apoB level during lipid-relating therapy. PMID- 12901520 TI - Effects of certain vasoactive peptides on pathogenesis of vascular restenosis. AB - OBJECTIVE: To investigate the effects of several vasoactive peptides on the development of arterial restenosis after balloon angioplasty. METHODS: In rat aortic artery restenosis model produced by denudation of aortic endothelia, we observed changes of endothelin (ET), angiotensin II (AII), calcitonin gene related peptide (CGRP) and adrenomedullin (Adm) in plasma and aorta with radioimmunoassay and expression of hypertension-related gene (HRG-1) with semi quantitative RT-PCR, and studied the effects of these peptides on intimal hyperplasia, intima/media ratio and MAPK activities of aortic artery after angioplasty respectively. Furthermore, in cultured cells, we studied the effects of these peptides on vascular smooth muscle cell (VSMC) proliferation and expression of HRG-1 of VSMC from spontaneously hypertensive rats (SHR) and Wistar Kyoto (WKY) rats with 3H-TdR incorporation and RT-PCR respectively. RESULTS: After angioplasty, the levels of ET and AII in plasma and aorta significantly increased, accompanied with VSMC proliferation and neointima hyperplasia. On day 10 after angioplasty, the levels of ET in plasma and aorta increased by 69% and 124% respectively, compared with sham group (P < 0.01); and the level of aortic AII increased by 80% (P < 0.01). Antiserum against ET or inhibitors of angiotensin converting enzyme (ACE) could significantly inhibit the proliferation of VSMC and neointima formation. Compared with the sham group, on day 3 after angioplasty, the CGRP levels in plasma and aorta increased by 64% and 89% respectively (P < 0.01) and the Adm levels in plasma and tissue increased by 129% and 102% respectively (P < 0.01). On day 10, intravenous administration of CGRP significantly inhibited the proliferation of VSMC and neointima formation induced by balloon aortic injury (by 66% and 79% respectively, P < 0.01). In addition, ET and AII attenuated the expression of HRG-1 in aorta and stimulated mitogen activated protein kinase (MAPK) activity, while CGRP and Adm potentiated the expression of HRG-1 and inhibited MAPK. CONCLUSIONS: ET and AII can stimulate the proliferation of injured intima while CGRP and Adm have an anti-hyperplasia effect after angioplasty. These 4 peptides are involved in the regulation of VSMC proliferation and affect the development of vascular restenosis by regulating the expression of HRG-1 and MAPK activity. PMID- 12901522 TI - Effects of HBV preS as a humoral enhancer on the abilities of HCV E2 protein to induce immune responses in the DNA-immunized mice. AB - OBJECTIVE: To study whether the abilities of hepatitis C virus (HCV) E2 gene immunization to induce humoral and cellular immune responses to E2 protein were affected by hepatitis B virus (HBV) preS gene when they fused in DNA-immunized mice. METHODS: Mice were immunized with E2, preS-E2 (preS gene was upstream of E2 gene), and E2-preS (preS gene was downstream of E2 gene) gene by their eukaryotic expression vectors, respectively. The anti-E2 or anti-preS antibodies were detected using the E2 and preS antigens. The cellular immune response to E2 protein in immunized mice was presented by its survival time after injecting SP2/O myeloma cells expressing HCV E2 protein into the abdominal cavity. RESULTS: Chimeric E2 and preS gene immunization can induce mice to develop anti-preS and anti-E2 antibodies. The number of the mice developing anti-E2 antibody and the antibody titers in preS-E2 gene-injected group were higher than those in E2-preS gene-immunized group. However, the mice injected with E2 gene did not develop the detectable anti-E2 antibodies until 12 weeks after DNA immunization. After the mice was injected with target cells, the average survival time of the mice in the group immunized with E2 gene alone was longer than that of the group injected with E2 gene fused with HBV preS and was significantly longer than that of the control (P < 0.05). CONCLUSION: HBV preS might be a humoral enhancer that can affect the abilities of HCV E2 protein to induce immune responses in DNA immunized mice. PMID- 12901523 TI - Occurrence of the lung fluke, Paragonimus heterotremus in Manipur, India. AB - OBJECTIVES: To determine the natural crustacean host, the Paragonimus species, and to investigate the potential host-parasite relationship between Manipur Paragonimus and some of the laboratory animals. METHODS: The laboratory animals such as puppies, albino rats and Swiss mice were infected orally with metacercariae isolated from the fresh water crabs, Potamiscus manipurensis. The fecal specimens of the experimentally infected animals were examined microscopically for Paragonimus eggs at regular intervals. The animals were autopsied on days 35 to approximately 328 after infection and the isolated worms were flattened between glass slides and fixed in 70% alcohol. The worms were stained with carmine and mounted with Canada balsam for morphological studies. The eggs were collected in 5% formol saline solution for microscopy. The flukes were classified into 4 developmental stages. RESULTS: A total of 11 worms, 5 mature, 5 immature and 1 pre-adult were recovered. The morphological features of the metacercariae, worms and eggs were similar to those of Paragonimus heterotremus. CONCLUSION: Manipur is one of the rare areas in the world where Paragonimus heterotremus is prevalent and the puppies are ideal experimental animal host. This species may be one of the important causes of paragonimiasis in animals and humans in Manipur. PMID- 12901524 TI - Detection of platelet-derived microparticles using flow cytometry and its clinical application. AB - OBJECTIVE: To establish a flow cytometric internal standard method for counting platelet-derived microparticles (PMPs) and to study its clinical significance. METHODS: PMPs suspension (platelet poor plasma, PPP) was extracted by gradual centrifugation. According to the size of PMPs, 3 microm and 0.8 microm latex beads were used as internal standards for the quantitation. PMPs were counted by adjusting flow cytometric discrimination and voltage of forward scatter and side scatter. RESULTS: In 30 healthy donors, the average concentration of resting PMPs was (1.2 x 10(5) +/- 5.7 x 10(4))/ml and that of activated PMPs was (1.6 x 10(6) +/- 9.1 x 10(5))/ml. Compared with healthy donors, PMPs mean value was significantly higher (P < 0.001) in 18 patients with coronary artery disease, 12 with acute cerebral infraction and 23 with chronic renal failure [the average PMPs concentration, (6.1 x 10(5) +/- 2.5 x 10(5))/ml, (6.8 x 10(5) +/- 3.4 x 10(5))/ml and (5.9 x 10(5) +/- 3.1 x 10(5))/ml respectively]. However, no significant difference in PMPs concentration was observed in 25 patients with acute leukemia and severe thrombocytopenia during the aplastic phase after chemotherapy [(1.3 x 10(5) +/- 6.1 x 10(4))/ml, (P > 0.05)]. CONCLUSIONS: PMPs is a useful indicator in monitoring platelet activation, and plays an important role in thrombotic disease. By flow cytometric internal standard method, PMPs can be counted rapidly and accurately, which may be very helpful in interlaboratory comparative studies. PMID- 12901525 TI - Single photon emission computed tomography study of regional cerebral blood flow in patients with hemispatial neglect. AB - OBJECTIVE: To explore the correlations between the occurrence and severity of neglect and the region, range or extent of the decrease in regional cerebral blood flow (rCBF). METHODS: Nineteen dextromanual patients who were diagnosed as unilateral stroke clinically and hemispatial neglect by a neglect test battery received single photon emission computed tomography (SPECT) scans. RESULTS: On images, the damages of patients with neglect were seen most frequently in the frontal cortex, and then in turn in the parietal cortex, occipital cortex, temporal cortex, basal ganglia and thalamus. Most patients with neglect had two or more regions damaged. The most significant region was temporal-parietal occipital (TPO) junction. The correlation coefficient between rCBF and the severity of neglect was -0.34 (t = -1.5, P > 0.05), and that between the decrease percentage of rCBF and the severity of neglect was 0.34 (t = 1.47, P > 0.05). The correlation coefficients between the range, number of foci, the flow deficit size and the severity of neglect were 0.71 (t = 4.13, P < 0.01), 0.70 (t = 4.07, P < 0.01) and 0.64 (t = 3.40, P < 1.01), respectively. CONCLUSIONS: The severity of neglect correlates with rCBF and the decrease percentage of rCBF insignificantly, but correlates positively with the range, number of foci and the flow deficit size significantly. Hemispatial neglect is caused by the damage of multiple sites and combined damage results in more severe neglect. PMID- 12901526 TI - MR imaging of meningeal carcinomatosis by systemic malignancy. AB - OBJECTIVE: To investigate the magnetic resonance (MR) features of meningeal carcinomatosis, and to improve the ability in understanding and diagnosing meningeal carcinomatosis by MR findings. METHODS: Eleven cases with proven meningeal carcinomatosis were studied by conventional and Gd-DTPA enhanced MR imaging. The enhancement patterns and features, as well as the types of meningeal involvement, were retrospectively analyzed. RESULTS: Conventional MR imaging showed no evident meningeal abnormalities. After the administration of Gd-DTPA, abnormal pia mater enhancement was detected in 9 cases, demonstrating as the continuous, thin, and lineal high signal intensity on the brain surface that could descend into the sulci. The abnormal pial enhancement occurred on the cortical surfaces of cerebellum, brainstem, and cerebrum. No abnormal enhancement in the subarachnoid space was found. Abnormal dura-arachnoid enhancement was seen in 3 cases, showing as the continuous, thick, and curvilineal high signal intensity over the convexities or in the tentorium without extension into the cortical sulci. Cerebral dura-arachnoid involvement was found in all 3 cases and one of them also showed abnormal enhancement in cerebellar dura-arachnoid and tentorium. Of the 11 cases, 9 with pial involvement had abnormal cerebrospinal fluid (CSF) results, 2 involving only the dura-arachnoid had normal CSF results. CONCLUSION: Meningeal carcinomatosis could be well demonstrated by Gd-DTPA enhanced MR imaging, and its type could be differentiated by the enhancement features. Combined with the clinical information, Gd-enhanced MR imaging may lead to the diagnosis and guide the therapy of meningeal carcinomatosis. PMID- 12901527 TI - Anterior spinal fusion with TSRH instrumentation for scoliosis. AB - OBJECTIVE: To introduce a new spinal internal fixation system, Texas Scottish Rite Hospital (TSRH), and to investigate its early clinical outcomes. METHODS: The preliminary clinical outcomes of 15 patients with thoracolumbar or lumbar scoliosis treated by anterior spinal fusion with TSRH instrumentation were studied retrospectively. Fourteen patients were diagnosed as idiopathic scoliosis and 1 as neuromuscular scoliosis. RESULTS: Preoperatively, the Cobb's angle on the coronal plane was 55.8 degrees (range 35 degrees to approximately 78 degrees), and 14 degrees postoperatively, with an average correction of 74.8%. The average unfused thoracical curve was 35.9 degrees preoperatively (range 26 degrees to approximately 51 degrees) and 21.8 degrees (10 degrees to approximately 42 degrees) postoperatively, with 40% correction. The sagittal curve of lumbar was kept physiologically, preoperative 27.9 degrees and postoperative 25.7 degrees respectively. The trunk shift was 13.4 mm (5 to approximately 28 mm) preoperatively and 3.5 mm (0 to approximately 7 mm) postoperatively. The averaged apic vertebra derivation was 47.8 mm (21 to approximately 69 mm) before operation and 10.8 mm (3 to approximately 20 mm) after operation. The distance of C7 to center sacrum vertical line (CSVL) was 19.5 mm (16 to approximately 42) preoperatively and 11.3 mm (0 to approximately 32 mm) postoperatively. The apical vertebra rotation was 3 degree in 15 patients preoperatively, and were improved to normal in 10 patients, 1 degree in 4 patients, and 2 degree in 1 patients postoperatively. None had neurological injury and infection. Only 1 patient complained a cool feeling in the low extremity of concave side, and the symptom vanished at 3-month followed up. CONCLUSION: If used appropriately, TSRH anterior spinal system is a good teatment for low thoracic or thoracic lumbar scoliosis. PMID- 12901528 TI - Pulmonary blood distribution after total cavopulmonary connection of different types. AB - OBJECTIVE: To assess the feature of pulmonary blood flow distribution after total cavopulmonary connection (TCPC) of different types, and to provide the selection of the best type. METHODS: Thirty-two consecutive survival patients after TCPC underwent radionuclide lung perfusion imaging. According to the radionuclide counts in the left and right lungs, analyses of the distribution of blood flow from superior venous cava (SVC) and inferior venous cava (IVC) and the whole pulmonary blood flow in both lungs were made. All patients were divided into 4 groups by the the anastomosis between IVC and pulmonary artery. RESULTS: Group I: The flow ratio of the IVC to left lung was greater than that to the right lung, P < or = 0.01; the flow ratio of the SVC to right lung was greater than that to the left lung, P < or = 0.01; and the whole pulmonary blood flow went dominantly to the left lung, P < or = 0.05, which is not in line with physiological distribution. Group II: the flows from the SVC and IVC were mixed in the middle of the junction and ran evenly into the right and left lungs, the whole pulmonay blood flow went to both lungs, P > or = 0.05. Group III: the flow ratio of the SVC to both lungs were the same, P > or = 0.05, and major part from IVC went to the right lung, P < or = 0.01; the pulmonary blood flow go dominantly to the right lung, P < or = 0.05, which is in accord with physiological distribution. Group IV: the flows from the right SVC went to right lung by 100%, P < or = 0.01, and that from the left SVC went to left lung by 100% too, P < or = 0.01; the flows from IVC went dominantly to the left lung, with little part to the right lung, P < or = 0.05. CONCLUSIONS: Different types of TCPC can result in different pulmonary blood distributions. The best flow distribution between the left and right lungs can be obtained by an offset of the IVC anastomosis toward the RPA with widening anastomosis for the patients without persist left superior venous cava (PLSVC). PMID- 12901529 TI - Change of carbon monoxide in plasma and tissue during acute hypoxia. AB - OBJECTIVE: To investigate the role of endogenous carbon monoxide (CO) in hypoxia. METHODS: After rats were inhaled with hypoxic gases and the heme oxygenase inhibitor ZnPPIX was administered, we measured the CO levels in plasma, liver, lung and kidney. Meanwhile plasma cGMP levels were observed. Furthermore, we recorded the change of hemodynamic and blood gases. RESULTS: Acute mild hypoxia (10% O2) significantly increaed CO levels in plasma as well as liver, kidney and lung, while acute severe hypoxia (5% O2) significantly decreased CO levels in plasma as well as liver, kidney and lung. In addition, the former significantly elevated cGMP levels in plasma while the latter markedly reduced cGMP levels in plasma. The hemodynamic change occurred in accordance with the changes carbon monoxide. CONCLUSION: Our results indicate, for the first time, that the endogenous carbon monoxide plays an important role in regulating the vessel tone during hypoxia. PMID- 12901530 TI - Clinical study of patient-controlled epidural analgesia with tetracaine hydrochloride after pulmonary lobectomy. AB - OBJECTIVE: To investigate the efficacy and safety of tetracaine hydrochloride in patient-controlled epidural analgesia (PCEA) after pulmonary lobectomy. METHODS: Forty-three patients scheduled for elective pulmonary lobectomy under general anesthesia were randomly allocated into either tetracaine group (22 patients) or ropivacaine group (21 patients). In the tetracaine group, 0.15% tetracaine was used for postoperative PCEA, while 0.3% ropivacaine was used in the ropivacaine group. The duration of postoperative analgesia was 48 h. The PCEA included a bolus of 6 ml with a lockout time of 1 h. Postoperative pain score was measured by visa analogue scale (VAS). Forced expired volume at the 1st second (FEV1.0), forced vital capacity (FVC), FEV1.0/FVC and peak expired flow (PEF) were measured preoperatively and daily after surgery. Hemodynamics were monitored and recorded before and after each administration of local anesthetics during the period of the study. RESULTS: VAS scores in both groups decreased significantly after a bolus injection of local anesthetics. There was no significant difference between the two groups in VAS either before or after the administration of PCEA. On the 1st and 2nd days after the operation, pulmonary function was reduced in both groups. However, there were no significant differences between the percentage of the changes of FEV1.0, FEV1.0/FVC and PEF in the two groups. There were also significant differences between the percentage of the changes of heart rate, mean arterial blood pressure and SpO2 after administration of local anesthetics. There was no significant difference in overall satisfaction with pain relief. CONCLUSIONS: The analgesic effect of 0.15% tetracaine is similar to that of 0.3% ropivacaine used in patient-controlled epidural analgesia after thoracotomy. No serious side effects were observed. PMID- 12901531 TI - Initial comparison of intracranial aneurysm embolization with mechanical detachable spirals and with Guglielmi detachable coils. AB - OBJECTIVE: To compare the embolization effects of intracranial aneurysm with mechanical detachable spirals (MDS) and with Guglielmi detachable coils (GDC). METHODS: One hundred and twenty cases with 125 intracranial aneurysms were embolized in Beijing Hospital from March 1995 to July 1999. Sixty-six aneurysms in 64 cases were embolised with MDS, 51 in 48 with GDC, and 8 in 8 with both MDS and GDC. Clinical data including sex, age, subarachnoid hemorrhage (SAH), Hunt & Hess grading, diameter and neck width of aneurysms, number and length of coils used per aneurysm, occlusive ratio, and complications were compared between MDS and GDC groups. RESULTS: MDS and GDC group were comparable (t-test or chi2-test, all P value > 0.10) in terms of age, sex, diameter of aneurysms [(8.46 +/- 3.42) mm vs. (7.38 +/- 3.45) mm], neck width [(3.49 +/- 1.50) mm vs. (3.26 +/- 1.52) mm], coils number [(4.65 +/- 3.01) vs. (4.24 +/- 2.65)] and their length [(460.2 +/- 398.5) mm vs. (422.9 +/- 387.1) mm] used per aneurysm, occlusive ratio in aneurysms embolized > or = 80% [(95.00% +/- 6.32%) vs. (94.19% +/- 7.63%)], mortality and permanent complications (7.8% vs. 4.2%). CONCLUSIONS: MDS and GDC are all materials for embolization of intracranial aneurysms. MDS is less expensive, but more difficult to control and of propensity to complications while GDC is more compliant, easier to be used, safer, and have many alternative types for use as well a more extensive indications. PMID- 12901532 TI - Endometriosis in adolescents--analysis of 6 cases. AB - OBJECTIVE: To study the characteristics, diagnosis and treatment of endometriosis in adolescent patients. METHOD: Six cases of adolescent endometriosis in our hospital were reviewed retrospectively. RESULT: Endometriosis is the most common cause of chronic pelvic pain in adolescents. There may be a natural progression of endometriosis from atypical lesions in adolescents to classic lesions in adults. Congenital abnormalities of the reproductive tract are the main reasons for the adolescent endometriosis. CONCLUSION: Endometriosis should be strongly suspected in adolescent girls with chronic pelvic pain, especially unresponsive to oral contraceptives and nonsteroidal anti-inflammatory drugs. The treatment involves the operations and medicines. PMID- 12901533 TI - The localization of adrenomedullin in rat kidney tissue and its inhibitory effect on the growth of cultured rat mesangial cells. AB - OBJECTIVE: To observe the localization of adrenomedullin (AM) in rat kidney tissue and its inhibitory effect on the growth of cultured rat mesangial cells (MsC). METHODS: A monoclonal antibody against AM developed by our laboratory was used to detect the localization of AM protein in rat kidney tissue by avidin biotin complex immunohistochemistry. The expressions of AM and its receptor CRLR mRNA on cultured glomerular epithelial cells (GEC) and MsC were investigated by Northern blot assay, and the possible effect of AM secreted by GEC on MsC proliferation was observed using [3H]thymidine incorporation as an index. RESULTS: A specific monoclonal antibody against AM was succesfully developed. AM was immunohistochemically localized mainly in glomeruli (GEC and endothelial cells), some cortical proximal tubules, medullary collecting duct cells, interstitial cells, vascular smooth muscle cells and endothelial cells. Northern blot assay showed that AM mRNA was expressed only on cultured GEC, but not on MsC, however, AM receptor CRLR mRNA was only expressed on MsC. GEC conditioned medium containing AM can inhibit MsC growth and AM receptor blocker CGRP8-37 may partially decreased this inhibitory effect. CONCLUSION: AM produced by GEC inhibits the proliferation of MsC, which suggests that AM as an important regulator is involved in glomerular normal physiological functions and pathologic processes. PMID- 12901534 TI - Effect of high-lipid diet on glomerular mesangial matrix in adriamycin-induced nephrotic rats. AB - OBJECTIVE: To determine the effect of hypercholsterolemia induced by a high-lipid diet on glomerulosclerosis. METHODS: Twenty nephrotic syndrome (NS) Wistar rats administrated adriamycin (ADR) with a single intravenous dose of 5 mg/kg body weight, were divided into the standard and high-lipid chow groups. Another 20 weight-matched non-NS rats that received a vehicle alone were grouped as control. Urinary protein excretion and serum cholesterol were assayed; image analysis and techniques of pathology, immunohistochemistry, and molecular biology were used to determine morphological changes in glomeruli and the production of glomerular mesangial matrices in different groups. RESULTS: The serum total cholesterol level was significantly higher in rats with high-lipid chow in both non-NS [(2.2 +/- 0.3) g/L vs. (0.9 +/- 0.1) g/L, P < 0.01] and NS [(9.5 +/- 0.2) g/L vs. (2.3 +/- 0.3) g/L, P < 0.01]. The urinary protein excretion was significantly higher in the high-lipid diet rats than in standard chow rats [(76.2 +/- 24.2) mg/24h vs. (44.8 +/- 13.6) mg/24h, P < 0.05] in NS rats. Although increases in the mesangial matrix and mesangial cells were observed in rats with high-lipid diet in both NS and non-NS group, more obvious pathological changes were found in NS group, such as lipid deposits and foam cell formation in mesangial areas, and progressing to focal and segmental glomerulosclerosis in some glomeruli. The immunohistochemical asay showed that the production of 3 major components (collagen IV, fibronectin, and laminin) was increased in NS group, especially in the rats with high-lipid chow. The increased expression of laminin mRNA was also detected with slot blotting in both NS and non-NS rats with high-lipid chow, and it was more obvious in the rats with NS. CONCLUSION: Our findings indicated that diet-induced hyperlipidemia can lead to over-production of mesangial matrix components, and further aggravate glomerulosclerosis in ADR-induced nephrosis. PMID- 12901535 TI - Construction, expression and biological assessment of BPI23-Fcgamma1 recombinant protein prokaryotic expression vector. AB - OBJECTIVE: To construct pBV-BPI600-Fcgamma1(700) recombinant expression vector, to transform it into Escherichia coli DH5alpha, and to induce the expression of BPI23-Fcgamma1 anti-bacterial recombinant protein. METHODS: Genes coding for BPI23 and Fcgamma1 were amplified by RT-PCR from mRNA extracted from HL-60 cell and normal human leukocytes; recombinant cloning vector and recombinant expression vector were then constructed. pBV-BPI600-Fcgamma1(700) recombinant expression vector was transformed into the competent Escherichia coli DH5alpha and BPI23-Fcgamma1 recombinant protein was expressed by a temperature-induced method. RESULTS: (1) Expected amplified products BPI600hp and Fcgamma1(700bp) were obtained by RT-PCR method. (2) pUC18-BPI180, pUC18-BPI420 and pUC18 Fcgamma1(700) recombinant cloning vector were successfully constructed, and sequences were identical with the reported ones. 3) pBV-BPI600-Fcgamma1(700) recombinant expression vector was successfully constructed, and the enzyme digestion analysis showed an expected result. (4) The expression level of BPI23 Fcgamma1 recombinant protein accounted for 20% of total bacterial proteins. (5) The renatured BPI23-Fcgamma1 recombinant protein showed bacteriocidal activity and biological function of complement fixation, and opsonization. CONCLUSION: pBV BPI600-Fcgamma1(700) recombinant expression vector was successfully constructed, and BPI23-Fcgamma1 recombinant protein with double biological activity of BPI and IgGFc was expresed in Escherichia coli. PMID- 12901536 TI - Expression of midkine in benign, premalignant and malignant vulvar tumors. AB - OBJECTIVE: To clarify the role of midkine (MK) in vulvar carcinogenesis though examination of its expression in vulvar lesions including vulvar condyloma acuminata (VCA), vulvar intraepithelial neoplasia (VIN) and vulvar squamous cell carcinomas (VSCC), and to analyze the relationship between MK expression and human papilloma virus (HPV) infection. METHODS: Thirty VSCC, 15 VIN and 10 VCA patients were studied by streptavidin-biotin-immunoperoxidase method. MK expression was compared with clinicopathologic features of vulvar tumors. RESULTS: MK was expressed in 26 of 30 VSCC (87%), 3 of 5 VIN III and all VCA samples, whereas no MK expression was detected in the VIN I-II samples or in normal epithelium. The difference of MK expression between VIN III and VSCC was statistically significant (P < 0.05). MK was more intensely expressed in differentiated-type (well differentiated and moderately differentiated) VSCC than in undifferentiated-type (poorly differentiated) VSCC. There was no statistically significant correlation between MK expression and clinical stage, lymph node metastasis and HPV infection in VSCC. MK expression were observed in all HPV positive specimens including 2 VSCC, 1 VIN III and all VCA. CONCLUSIONS: MK gene expression may be a late event in vulvar squamous cell malignant transformation, and may be associated with vulvar tumor cell differentiation. HPV-positive vulvar tumors expressed MK protein. PMID- 12901537 TI - Study on the anti-tumor efficacy induced by heat shock protein 70-peptide complexes derived from tumor cells. AB - OBJECTIVE: To study the efficacy and explore the mechanism of the anti-tumor immunity elicited by heat shock protein 70-peptide complexes (HSP70-PC) derived from tumor cells. METHODS: Cells culture, flow cytometric analysis, affinity chromatography for protein purification, SDS-PAGE, Western-blotting and animal experiment were used. RESULTS: HSP70-PC immunization rendered protective effect to both naive tumorl-bearing mice. All of the naive mice obtained complete resistance to Hcaf cell attack; 40% of the tumor-bearing mice survived for over 90 days, whereas the mice of control group died within 2 weeks (P < 0.01). CD8+ subset of T lymphocytes in the peripheral blood of immunized mice increased by 12%. CONCLUSION: HSP70-PC induces anti-tumor immunity via activation of cytotoxic T lymphocytes (CTLs), and it possesses strong tumor vaccine effect. Our research adds more evidence to support the clinical use of HSP70-PC to fight human cancers. PMID- 12901538 TI - Expression of rhBMP-7 gene in transduced bone marrow derived stromal cells. AB - OBJECTIVE: To explore the possibility of expression of exogenous gene in transduced bone marrow derived stromal cells (BMSCs). METHODS: The marker gene, pbLacZ, was transferred into cultured BMSCs and the expression of transduced gene by X-gal staining was examined. Then plasmid pcDNA3-rhBMP7 was delivered to cultured BMSCs. Through immunohistochemical staining and RT-PCR assay, the expression of rhBMP7 gene was detected. RESULTS: The exogenous gene could be expressed efficiently in transduced BMSCs. CONCLUSION: The present study provided a theoretical basis to gene therapy on the problems of bone and cartilage tissue. PMID- 12901539 TI - Diagnostic value of serum insulin-like growth factor binding protein-3 in children with or without growth hormone deficiency. AB - OBJECTIVE: To study the value of serum insulin-like growth factor binding protein 3 (IGFBP-3) levels in differential diagnosis of growth hormone deficiency (GHD). METHODS: To measure serum IGFBP-3 levels by RIA in normal children and adolescents, GHD children and short-stature children without GHD. RESULTS: Serum level of IGFBP-3 in 129 children with untreated GHD and with no pubertal development was 1.6 +/- 0.9 mg/L, which was less than that in normal group of the same age, but overlapped with the normal children in Tanner stage I. After six month treatment with recombinant human growth hormone (rhGH), serum level of IGFBP-3 in 59 GHD significantly increased from 1.3 +/- 0.7 mg/L to 2.7 +/- 0.9 mg/L, accompanied by an increase of body heights, growth velocities and serum level of IGF-1. Serum level of IGFBP-3 in 55 short-stature children without GHD was 3.3 +/- 2.2 mg/L, which was not significantly different from that in normal group. CONCLUSION: Serum IGFBP-3 level can reflect the status of GH secretion in children with GHD and is a useful marker for differential diagnosis of GHD. PMID- 12901540 TI - Sonographic patterns and differential diagnosis of cystic renal carcinomas. AB - OBJECTIVE: To study the sonographic features and patterns of cystic renal carcinomas. METHODS: Thirteen cases of cystic renal carcinoma confirmed by operation and pathology were examined by ultrasonography, and the cystic walls, septa and solid mural nodules were studied. RESULTS: Solid mural nodules of some cases and irregular thickening of the cystic walls and septa were characteristic findings for the ultrasonic diagnosis of cystic renal carcinomas. According to their pathologic mechanisms and sonographic features, cystic renal carcinomas were classified into 3 patterns: unilocular cystic mass, multiloculated cystic mass and cystic-solid mass. CONCLUSIONS: Typical cystic renal carcinomas can be well diagnosed, while atypical cases may be misdiagnosed as benign renal cysts by ultrasonography. Color Doppler ultrasonography and needle aspiration guided by ultrasonography are helpful in the diagnosis of these atypical cases. PMID- 12901541 TI - Clinical comparison of the selective serotonin3 antagonists ramosetron and granisetron in treating acute chemotherapy-induced emesis, nausea and anorexia. AB - OBJECTIVE: The efficacies of the selective 5-hydroxytryptamine3 (5-HT3) antagonists--ramosetron (0.3 mg) and granisetron (3 mg) in treating acute chemotherapy-induced digestive system dysunction were compared. METHODS: A total of 111 patients were enrolled in a single-blind, randomised crossover study; with data from 98 were used to assess efficacy and data from 110 to assess the safety profile. Ramosetron or granisetron was given intraveneously 15 min befire chemotherpy. RESULTS: The ability of ramosetron to prevent emesis, nausea and anorexia was similar to granisetron during the first 6 h following the administration of chemotherapy, ciplatin or doxorubicin. However, during the first 24 h after chemotherapy, significant differences between ramosetron and granisetron appeared: emetic episode (P = 0.068), nausea (P = 0.006), and anorexia (P = 0.048) remained lower in ramosetron-treated patients. The safety profile of ramosetron was similar to that of granisetron and adverse events in both groups were generally mild and transient. CONCLUSION: Ramosetron is more potent and longer-lasting than granisetron in preventing chemotherapy-induced digestive disturbances. PMID- 12901542 TI - Diagnosis and surgical management for retrosternal thyroid mass. AB - OBJECTIVE: To understand the clinical manifestations, diagnostic methods, surgical management and prognosis of retrosternal thyroid masses in various pathological types. METHODS: Sixty-four cases of retrosternal thyroid masses with surgical intervention were analyzed retrospectively. RESULTS: Trachea-compressed symptoms (65%) and shadows beside the trachea at thoracic inlet (94%) were the most common clinical findings, chest X-ray (70%) and CT scan (96%) had higher diagnostic rate. No death occurred during operation or hospitalization among these patients. Total complications occurred in 15.7% cases (11/70) (including 3 preoperative cases with hoarseness) and postoperative pathological results were mainly multinodular goiter (54.7%), thyroid adenoma (21.9%) and thyroid carcinoma (15.6%) (including local carcinomatous change). CONCLUSIONS: Diagnosis of retrosternal thyroid mass can be correctly made by chest X-ray and CT scan. Most operations on retrosternal thyroid masses can be performed safely through cervical incision with minimal morbidity and low recurrence rate. Retrosternal thyroid carcinoma was potentially invasive and could hardly be resected completely, hence with poor prognosis. PMID- 12901543 TI - Surgical management of Ehlers-Danlos syndrome: first report of a pedigree in China. AB - OBJECTIVE: To describe a case of Ehlers-Danlos syndrome type IV and its pedigree in China. METHODS: Clinical materials of a case of Ehlers-Danlos syndrome type IV and a pedigree of 6 members within 4 generations were analyzed. Dilated internal jugular vein in the proband was removed by operation. The diagnosis, surgical treatment, and postoperative complications were retrospectively reviewed. RESULTS: Vessels of the proband in the pedigree were crisp and easily lacinated during the procedure of removing his internal jugular vein. Repeating postoperative hematomas were found though complete stanching was achieved during the operation. The patient was successfully recovered by promptly debridgement and needle sucking. The other 5 members of the pedigree all had the triads of Ehlers-Danlos syndrome. CONCLUSIONS: Though it was of extremely low morbidity rate, the high mortality rate and complication of Ehlers-Danlos syndrome deserve great attention during surgical management, especially in patients with Ehlers Danlos syndrome type IV. Surgeons should be aware of the ponderance of its complications and combined diseases to avoid fatal intraoperative vascular lascination and incontrollable hemorrhage. PMID- 12901544 TI - Surgical treatment of metastatic spinal tumor. AB - OBJECTIVES: To evaluate the effect of surgical treatment on metastatic spinal tumor. METHODS: The results of surgical intervention for metastatic spinal tumor of 31 consecutive patients since October 1985 were reviewed. RESULTS: The average survival time was 17.6 months (range from 3 months to 9 years), and 4 patients are still alive with an average survival time of 24.6 months (range, 14 to approximately 84 months). No postoperative complication was noted. The preoperative symptoms were partially relieved and neurological functions were improved after surgery. CONCLUSION: Surgical treatment for metastatic spinal tumor could improve the life quality, but should be adopted cautiously. The surgical procedures such as decompression and internal fixation should be involved only when neurological deficits occurred. The surgery with postoperative complementary therapy may not only improve the life quality, but also extend the patients' life span. PMID- 12901545 TI - Characteristics of lip-mouth region in smiling position from 80 persons with acceptable faces and individual normal occlusions. AB - OBJECTIVE: The characteristics of lip-mouth region including the soft and hard tissues in smiling position with frontal fixed position photographic computer aided analysis were studied. METHODS: The subjects were 80 persons (40 male and 40 females, age range: 17 to approximately 25 years) with acceptable faces and individual normal occlusions. The subjects were asked to take maximum smiling position to accept photographic measurement with computer-aided analysis. RESULTS: The maximum smile line could be divided into 3 categories: low smile line (16.25%), average smile line (68.75%), and high smile line (15%). CONCLUSION: The method adopting maximum smiling position to study the lip-month region is reproducible and comparable. This study would be helpful to provide a quantitative reference for clinical investigation, diagnosis, treatment and efficacy appraisal. PMID- 12901546 TI - Wound infection after scoliosis surgery: an analysis of 15 cases. AB - OBJECTIVE: To discuss the causes and treatments of wound infections after scoliosis surgery. METHODS: Nine hundred and twenty-four caes of scoliosis were reviewed, and the clinical data of 15 cases of postoperative infection were analysed retrospectively. RESULTS: All 15 cases underwent spinal posterior fusion with autologous bone graft using instrumentations. Seven were diagnosed as early infection, and 8 were delayed infection. Radical debridement was performed in all 15 cases. The duration of antibiotics administration was 10 to 34 days with continuous closed irrigation for 2 to approximately 4 weeks and primary closure for the wounds. All patients were followed up for an average of 3.5 years (2 to 7.5 years) with good outcomes and no recurrence. CONCLUSION: Wound infection following surgical correction of scoliosis primarily results from intraoperative seeding, although host-related and operation-related factors may contribute to its development. Once the infections are diagnosed, good results can be achieved by prompt surgical debridement, irrigation and reasonably administered antibiotics. Removal of hardware may be necessary in deep infections. PMID- 12901547 TI - Association of HLA-DRB1 alleles with polymyositis/dermatomyositis in northern Chinese Hans. PMID- 12901548 TI - Vaccines: exploiting the role of toll-like receptors. PMID- 12901549 TI - AIDS vaccine 'within 5 years'. PMID- 12901550 TI - Cancer vaccines: unanswered questions. May 16, 2002, Phacilitate Spring Vaccine Forum, Cancer Vaccines, Paris, France. PMID- 12901551 TI - Gene-based vaccines: mechanisms, delivery systems and efficacy. April 10-15, 2002, Beaver Run Resort, Breckenridge, Colorado, USA. PMID- 12901552 TI - Recombinant hepatitis B triple antigen vaccine: Hepacare. AB - Infection with hepatitis B virus is a public health problem throughout the world. Hepatitis B vaccines are now included in national immunization programmes of infants and/or adolescents in 129 countries. Current single antigen vaccines, that are plasma-derived or produced by recombinant DNA technology are highly effective, but between 5-10% or more of healthy immunocompetent subjects do not mount an antihepatitis B surface antibody protective response and others respond poorly (hyporesponders). The inclusion of pre-S1 and -S2 components of hepatitis B surface antigen in addition to the single antigen (triple antigen) in a novel vaccine, Hepacare, Medeva Pharma Plc, Speke, UK, overcomes nonresponsiveness and hyporesponsiveness in a significant number of individuals. The triple antigen is indicated for vaccination of nonresponders (and hyporesponders) to the current single antigen vaccines and for persons who require rapid protection against hepatitis B infection. PMID- 12901553 TI - MMR and autism: moving from controversy toward consensus. AB - As controversy swirls around the putative association between the measles-mumps rubella vaccine and autism, the methods of scientific investigation, established more than 150 years ago, bear remembering. In this article, we will discuss the origins of this hypothesis, evidence in support of and at variance with this theory and future investigations that may help to resolve this debate. PMID- 12901554 TI - Vaccination against measles: a neverending story. AB - Measles, a highly contagious viral disease, is a major childhood killer in developing countries, accounting for almost 1 million deaths every year globally. Measles virus normally does not cause a persistent infection, no animal reservoir for measles virus exists, no vector is involved in its spread, only one serotype exists, the virus is antigenically stable and vaccination with the currently used live attenuated vaccines proved to be highly effective in preventing disease. Therefore, theoretically measles should be considered eradicable. This article provides a review of past and current measles vaccination efforts and development and need of new generation experimental measles vaccines. PMID- 12901555 TI - Methods of ensuring vaccine safety. AB - With a decreasing disease burden from vaccine-preventable diseases in developed countries, ensuring the safety of vaccines is critically important for maintaining public confidence in vaccination programs and continuing low levels of disease. This will require investment in resources to identify true adverse effects, understand their pathophysiology, and design and manufacture safer vaccines. PMID- 12901556 TI - DNA-based vaccines for allergic disease. AB - Allergen-specific immunotherapy, although efficacious, is now less frequently used because of potential adverse reactions. Recently, two new types of allergen immunotherapy have been developed that appear to overcome this problem, namely allergen gene vaccination and vaccination with allergen-immunstimulatory DNA conjugates. In animal models of allergy, both have been shown to induce nonallergic T-helper cell type 1 immune responses to allergens and downregulate pre-existing T-helper cell type 2 responses. In initial clinical trials with allergic patients, allergen-immunostimulatory DNA conjugates were well-tolerated, induced immunoglobulin-G but not immunoglobulin-E antibodies and appeared to have great potential as a novel, safe and efficacious type of allergen specific immunotherapy. PMID- 12901557 TI - What information is needed to design effective vaccination against intracellular pathogens causing chronic disease? AB - An appreciation of global features of immune regulation may lead to vaccination strategies effective in a genetically diverse population against a number of intracellular pathogens that cause chronic disease. Such global strategies appear more straightforward than strategies requiring a detailed knowledge of the specificity of 'protective T-cells'. Global strategies may be effective against the virus, bacteria and protozoa that respectively cause AIDS, tuberculosis and the leishmaniases. PMID- 12901558 TI - The case for polyvalent cancer vaccines that induce antibodies. AB - Antibodies are ideally suited for eradicating pathogens from the bloodstream and early tissue invasion. With regard to cancer cells, passively administered and vaccine induced antibodies have accomplished this, eliminating circulating tumor cells and systemic or intraperitoneal micrometastases in a variety of preclinical models. A series of cancer cell-surface differentiation antigens have now been identified and synthesized. Antibodies against each can be induced in most vaccinated patients using keyhole limpet hemocyanin conjugate vaccines with a potent immunological adjuvant. Polyvalent vaccines may be required due to tumor cell heterogenely, heterogeneity of the human immune response and the correlation between overall antibody titer against tumor cells and antibody effector mechanisms. Trials testing the clinical impact of these polyvalent vaccines in the adjuvant setting are planned for the near future. PMID- 12901559 TI - Polytope vaccines for the codelivery of multiple CD8 T-cell epitopes. AB - Vaccines against a number of diseases, including HIV, Epstein Barr virus, malaria and several cancers, are believed to require the coinduction of multiple alphabeta CD8+ cytotoxic T-lymphocyte responses that are directed towards a number of different target antigens. The difficulties associated with making large recombinant vaccines that contain numerous antigens has led to the development of alphabeta CD8+ cytotoxic T-lymphocyte polyepitope or polytope vaccine approach, where multiple (usually 8-10 amino acids long) alphabeta CD8+ cytotoxic T-lymphocyte epitopes, derived from several antigens are conjoined into single artificial constructs. Such polytope constructs can be delivered using a number of different vaccine vector modalities with each epitope in the construct emerging as individually immunogenic. PMID- 12901560 TI - Phagocytosis of synthetic particulate vaccine delivery systems to program dendritic cells. AB - Therapeutic prospects of particulates are increasingly recognized for vaccination purposes. Compared with biologic particulates, such as live or attenuated bacterial vectors and viral vectors, synthetic particulates may be expected to ease the hurdles of quality assurance and validation in vaccine development and production and shorten the time for approval and to the market. The ability of synthetic antigen-loaded particulates to elicit strong immune responses, even with low amounts of antigen and to weakly immunogenic epitopes, is suggested to be due to their efficient cross-talk with the most potent antigen-presenting cells, such as dendritic cells. Moreover, the potential of particulates for intracellular delivery and directing intracellular trafficking of antigens has evolved as a promising opportunity to target the major histocompatibility complex I pathway. In summary, synthetic particulate vaccine delivery systems are likely to play an increasingly active role in enhancing or even enabling the immunostimulating effect of antigens upon direct interaction with the target cells. PMID- 12901561 TI - Can nonliving nasal vaccines be made to work? AB - Nasal vaccines consisting of nonliving particulate formulations can induce immune responses of importance for protection against infection. The most promising results have been obtained with vaccines against influenza, pertussis and group B meningococcal disease. So far, however, the results do not challenge the standing of corresponding injectable vaccines, although results of experiments in animals do indicate that effective nonliving nasal vaccines may soon be developed. This will depend on refined immunization schedules to benefit from immunological memory and on formulations to make the vaccines more accessible to the immune system by way of mucosal adjuvants or immune modulators. PMID- 12901562 TI - Expression of immunomodulating molecules by recombinant viruses: can the immunogenicity of live virus vaccines be improved? AB - Several obstacles exist for the development and use of live attenuated vaccines, including difficulty in achieving a proper balance between attenuation of viral replication and immunogenicity; inducing a strong T-helper 1 response in early life when the immune system is T helper 2 biased and immunization is sometimes associated with immunopathology and the immunosuppressive effect of maternal antibodies in infants. For some viral infections, the immune response to natural infection does not confer solid protection, complicating the task of vaccine development. The development of methods for generation of recombinant viruses provided new opportunities for improving the immunogenicity of live virus vaccine candidates, including the construction of viruses that express cytokines or other immunomodulating molecules. Depending on the choice of immunomodulating molecule, various stages of the immune response can be affected, such as antigen presentation or T-cell proliferation and differentiation. In addition to using the approach for development of viral live attenuated vaccines, it is currently being explored for the development of antitumor vaccines. For this type of vaccine, expression of tumor antigens and one or more immunomodulating molecules by one or several recombinant viruses has been proposed. PMID- 12901563 TI - Vaccine safety activists on the Internet. PMID- 12901564 TI - Multistrain HIV vaccine. PMID- 12901565 TI - Cancer vaccines: is the future now? AB - Cancer immunotherapy is an area with a tremendous amount of promise. The field has matured in that the approaches and antigens appear very specific and highly reasonable. Furthermore, the pre-clinical models support their evaluation. In fact, the human immune responses against many of the vaccines are approaching thresholds that should start to impact disease. While the creativity and progress in technology was clearly evident, there is much work left to pursue. Clear evidence for this situation is reflected the low number of products and approaches that are in late-stage trials. This again reflects that many apparently promising technologies have trouble making the transition from mouse to human. However, the success in the antibody field strongly supports that once understood, the correct immunization strategies and antigens will have dramatic impact on disease. One improvement would be some optimization of approaches at least for immunogenicity in nonhuman primates. Another important issue for standardization, is to focus on the use of quantitative immune measurement systems rather than a reliance on bulk assay systems for decision-making. Such a minor change would allow more standardization of data interpretation across the field and allow for more rapid and intuitive vector evaluation, even if trials were not head to head. In this regard, the development of class II tetramer technology to the evaluation of tumor antigens where available would be a welcome advance. Furthermore, in general, flow-based assays for immune evaluation are similarly gaining acceptance. It is important to remember that the success of recent antibody products for lymphoma and breast cancer did not happen overnight. Rather, those successes were 20 years in the making. In this regard, for cancer immunotherapy, which is T-cell based, this similar anniversary is not yet upon us but it is getting close. The next few years are critical to this emerging field and likely to be full of promise, surprises and even success. PMID- 12901566 TI - Progress in vaccination against cancer (PIVAC) 2002. PMID- 12901567 TI - Needle-free epidermal powder immunization. AB - Due to the presence of a network of antigen-presenting cells and other cells with innate and adaptive immune functions, the skin is both a sensitive immune organ and a practical target site for vaccine administration. A handful of needle-free immunization technologies have emerged in recent years that aim to take advantage of these characteristics. Skin delivery technologies provide potentially safer alternatives to needle injection and promises increased efficacy in the prevention and/or therapy of infectious diseases, allergic disorders and cancer. In this review, we will cover advances in needle-free skin vaccination technologies and their potential applications to disease prevention and therapy. Emphasis will be placed on epidermal powder immunization and particle-mediated ('gene gun') DNA immunization, which use similar mechanical devices to deliver protein and DNA vaccines, respectively, into the viable epidermis. PMID- 12901569 TI - T-cell vaccination for autoimmune diseases: immunologic lessons and clinical experience in multiple sclerosis. AB - Autoreactive T-cells are regulated under the normal conditions and play an important role in autoimmune pathologies when they are dysregulated as a result of genetic, environmental and other unknown factors associated with various autoimmune diseases. The immune regulation of autoreactive T-cells may be regained by activating the regulatory network, such as the idiotype anti idiotypic network. Immunization with inactivated autoreactive T-cells (T-cell vaccination) can be used as a powerful means of activating the idiotype anti idiotypic network to deplete specific subsets of autoreactive T-cells potentially involved in autoimmune conditions. It induces regulatory immune responses that closely resemble the in vivo situation, where the immune system is challenged by clonal activation and expansion of given T-cell populations in various autoimmune diseases. Recent clinical trials in multiple sclerosis have begun to reveal the role of T-cell vaccination in the understanding of in vivo regulation of autoreactive T-cells and in the development of effective therapeutic strategies for multiple sclerosis and other autoimmune conditions. In this article, we will review the recent advances in T-cell vaccination in relationship to the regulatory mechanism induced by T-cell vaccination and the potential of T-cell vaccination as a treatment for T-cell-mediated autoimmune diseases. Current issues and thoughts related to the preparation of T-cell vaccine, the relevant sources of autoimmune T-cells and epitope spreading are also discussed. PMID- 12901568 TI - Immunogenicity and reactogenicity of DTPw-HB/Hib vaccine administered to colombian infants after a birth dose of hepatitis B vaccine. AB - OBJECTIVE: To evaluate, in an open study, the immunogenicity, safety and reactogenicity of a birth dose of hepatitis B vaccine followed by a three-dose course of diphtheria-tetanus whole-cell pertussis-hepatitis B vaccine, extemporaneously mixed with Haemophilus influenzae b (Hib) vaccine. METHODS: At 2, 4 and 6 months of age, a single group of 120 Colombian infants were enrolled in this study to receive a regimen consisting of three doses of the combination vaccine following a dose of hepatitis B vaccine at birth. RESULTS: Seroprotection/vaccine response rates to all vaccine antigens was 98-100% 1 month after completion of the full vaccination course. The vaccine had an acceptable reactogenicity profile and the incidence of reported local and general symptoms decreased with the administration of subsequent vaccine doses. CONCLUSION: The mixed DTPw-HB/Hib vaccine was safe and well-tolerated, with high immunogenicity against all component antigens. Compared with previous studies, reactogenicity did not increase with the additional dose of hepatitis B vaccine given at birth. The DTPw-HB/Hib combination can be used to provide primary vaccination of infants who have already received a first dose of hepatitis B vaccine at birth. PMID- 12901570 TI - Conditional live virus as a novel approach towards a safe live attenuated HIV vaccine. AB - To control the worldwide spread of HIV, a safe and effective prophylactic vaccine is urgently needed. Studies with the simian immunodeficiency virus demonstrated that a live attenuated virus can be effective as a vaccine, but serious concerns about the safety of such a vaccine virus have arisen. We propose a conditional live virus, of which the replication can be switched on and off at will, as a novel approach for an HIV vaccine. PMID- 12901571 TI - Cellular vaccine therapy for cancer. AB - Observations that cells of the immune system are able to kill tumor cells both in vitro and in animal models have provided a compelling rationale for pursuit of a strategy whereby immune cells are administered as a therapeutic vaccine to patients with cancer. The successful outcome of this approach depends upon the ability to deliver this therapy in a manner in which a potent immune response is elicited. By harnessing the capacity of dendritic cells that are pivotal in priming the immune response and using gene therapy approaches to optimise the immune response, this may ultimately prove efficacious in the management of human cancer. Promising reports from recent clinical trials suggest that this may well be a realistic goal. PMID- 12901572 TI - Exoerythrocytic malaria vaccine development: understanding host-parasite immunobiology underscores strategic success. AB - Malaria imposes an enormous health burden on people living in the tropics and effective control measures are urgently needed. The vast majority of deaths in humans from malaria are caused by one species of the protozoan, Plasmodium falciparum. An efficacious and cost-effective vaccine against this parasite is considered a holy grail of modern molecular medicine. A vaccine that targets liver-stage parasites would prevent infection from reaching the blood and causing clinical disease. Among around 40 known Plasmodium falciparum antigens, only a few are expressed exclusively by mosquito-transmitted sporozoites or infected hepatocytes. Studies in humans have consistently related immune responses to these antigens with resistance to infection or disease, providing a powerful rationale for the development of pre-erythrocytic vaccines. By dissecting the mechanism(s) of immunity to these antigens, we can best evaluate in different delivery systems epitopes associated with protection as components of a focused and coordinated multiantigen malaria vaccine strategy. PMID- 12901573 TI - Tuberculosis vaccines: the past, present and future. AB - Tuberculosis still remains a leading infectious cause of death worldwide, although the BCG vaccine has been used for 80 years. There is an urgent need to develop improved BCG or new tuberculosis vaccines. This apparently represents a daunting task, since it will take a long time before a vaccine can be declared to be better than the current BCG vaccine, both in experimental and human studies. The current review takes a brief historic look at the use of current BCG vaccine and provides an overview on what are considered to be the key immunologic criteria that have to be met by a new generation of tuberculosis vaccines. It also provides the most up-to-date information on the latest developments in tuberculosis vaccine research, with a focus on mycobacterial organism-based and Mycobacterium tuberculosis antigen-based vaccines. Consideration is also given to the mucosal route of immunization and 'prime and boost' regimens. This review also presents several important tables, highlighting critical components of antituberculosis immunity, the most commonly tested immune adjuvants, the types of novel tuberculosis antigen-based vaccines and the outcome of different heterologous 'prime and boost' vaccination regimens. PMID- 12901574 TI - Will the current measles vaccines ever eradicate measles? AB - Measles virus is the most infectious transmissible agent causing human disease and has probably been responsible for the deaths of more children than any other single cause. In addition, infection with the natural virus causes many severe complications, including encephalitis, deafness and pneumonia. The introduction of live attenuated vaccines, either singly or as the measles-mumps-rubella combined vaccine, has dramatically reduced the occurrence of disease and in countries where vaccine uptake is high, indigenous disease has been virtually eliminated. Even though the current vaccines are very efficient, they do have their limitations. Children are most at risk during the first year of life and for most of this period, maternal antibodies prevent effective immunization. In addition, the current measles vaccines are relatively heat labile which causes difficulty in tropical areas. In recent years, vaccination rates in some industrial countries have been adversely affected by fears that measles vaccines are linked to inflammatory bowel diseases and autism. Although there is no conclusive evidence to support these fears, they still remain and probably contribute to poor vaccine uptake in some regions and sections of society. Although severe complications from vaccination are extremely rare, mild local reactions are more common. Consequently, in countries where measles is declining or has been eliminated, the fear of side effects of vaccination may encourage the development of vaccines that do not rely on virus replication to take effect. PMID- 12901575 TI - Therapeutic vaccines for substance dependence. AB - Several immunotherapies are under development for nicotine, cocaine and phencyclidine and a cocaine vaccine has started human trials. These therapies promise a new approach to diseases that have had limited treatment success and tremendous morbidity. Both the cocaine and nicotine addiction immunotherapies have reduced 'relapse' to drug use in animal model systems. To date, the active cocaine vaccine has few side effects and induces considerable antibody titers after active immunization in humans. Studies with the monoclonal phencyclidine immunotherapy provide intriguing evidence of sustained protection for months after single-dose administration. Other immunotherapy may include treatment of drug overdose, prevention of brain or cardiac toxicity and protection of a fetus during pregnancy in a drug abuser. PMID- 12901576 TI - Vaccines against hantaviruses. AB - Rodent-borne hantaviruses are etiologic agents for hemorrhagic fever with renal syndrome and hantavirus cardiopulmonary syndrome in the Old World and New World, respectively. These often severe diseases are relatively uncommon in most parts of the world and are sufficiently genetically variable that widely cross protective vaccines will probably need to be polyvalent. The current status of hantavirus vaccines shall be reviewed and both conventional and speculative new vaccine technologies that may evolve within the field shall be considered. PMID- 12901577 TI - Development of oral microencapsulated forms for delivering viral vaccines. AB - Rapid development in biotechnology during the last decade has allowed novel ideas in the development of antiviral vaccines to be considered and provides interesting technological approaches to their realization. Designing of microencapsulated forms for delivering bacterial and viral antigens or antigenic complexes using biodegradable biopolymers is an important novel direction. This approach involves the production of polymeric spherical particles with a diameter of 1 microm to 3 mm, containing isolated viral antigens or whole viral particles. Microencapsulated antigens administered orally are protected from low pH values of the gastric juice, bile acids, their salts and proteolytic enzymes of the gastrointestinal tract. The ability to drastically potentiate the immune response to encapsulated antigens, together with the ability to penetrate into the intestinal and respiratory mucosae upon oral and tracheal administrations, respectively, with induction of local and systemic immune reactions are the special merits of such polymers. However, the majority of data on microencapsulated viral vaccines has so far been obtained in animal models, as well as a limited number of studies on the protective effect they elicit. Certain success in the development of vaccines against a number of human viral infections, such as hepatitis B, cytomegalovirus and rotavirus, gives hope to successful completion of this research. Presumably, such vaccines will be safe and innocuous, simple in administration and capable of inducing both the systemic and local immune responses at the primary portal of viral infection. PMID- 12901578 TI - Evolution of conjugate vaccines. AB - Conjugate vaccines--bacterial polysaccharides conjugated to proteins to improve their immunizing properties--have been a success story since their introduction less than 20 years ago. The Haemophilus influenzae type b conjugate vaccine has nearly eliminated invasive Haemophilus influenzae type b disease in large parts of the world. The key reasons for its success are its ability to induce immunologic memory and reduce asymptomatic carriage, hence the spread of infection. The first pneumococcal conjugate vaccine was licensed in 2000 and is already in wide demand in the USA. The first meningococcal conjugate vaccine was introduced in a nationwide program in the UK in 1999. Current discussion focuses on the efficacy of the conjugate vaccines for different end-points and the cost effectiveness of their use globally. PMID- 12901579 TI - Population control: are contraceptives the only panacea? PMID- 12901580 TI - PREPS and L-particles: a new approach to virus-like particle vaccines. AB - Conventional virus-like particles are usually composed of a single structural protein which spontaneously assembles into particles. L-particles, a little-known type of virus-like particle, are produced as part of the natural infectious process of many, if not all, alpha-herpesviruses. L-particles lack the nucleocapsid present in the infectious virion but contain all of the virus envelope and tegument proteins. L-particles contain no virus DNA and are noninfectious, though they are biologically competent, since they are capable of delivering viral envelope and tegument proteins to cells. When cells are infected with herpes simplex virus Type 1 under conditions where viral DNA synthesis is blocked, previral DNA replication enveloped particles are produced. These are similar to L-particles, but differ slightly in protein composition. This article reviews the available data regarding these vaccine candidates and explores the wide-ranging potential applications, including vaccine candidates against infectious diseases and cancer, as well as a protein delivery vector. PMID- 12901582 TI - Current advances in antifertility vaccines for fertility control and noncontraceptive applications. AB - The search for effective antifertility vaccines has been actively pursued for decades in clinical applications as contraceptives or in the treatment of hormone dependent conditions. More recently, innovations have seen immunocontraception being employed successfully in veterinary and farming applications, as well as in population control of a number of different wildlife species. Although the basic principle behind these vaccines is the same, design considerations and criteria for commercialization are very different. This review will examine current advances in this field and evaluate the future scope of antifertility vaccines. PMID- 12901581 TI - Emerging issues in vaccine economics: perspectives from the USA. AB - Economic studies of vaccines, including vaccine development and delivery issues, are increasingly needed to inform policy recommendations and programmatic decisions in the USA. This need arises from the increasing costs of vaccines, the complexity of the US healthcare system and the limited number of vaccine manufacturers in the market. We have developed a national research agenda in domestic and global vaccine economics by conducting key informant interviews with 42 experts and inviting ideas from an additional 128 experts. To assess priorities among the 129 ideas that were generated, we asked 15 experts representing a broad range of perspectives to rank the ideas and we analyzed their votes. The highest-ranking domestic research ideas included evaluating: the costs of vaccine shortages, the cost-effectiveness of potential human papillomavirus vaccination and adult and adolescent pertussis vaccination programs and the cost-effectiveness of universal vaccine purchase programs for adults as well as children. The highest-ranking globally-oriented ideas included developing a resource allocation model to support the best vaccination program decisions with limited funds and assessing the cost-effectiveness of HIV, rotavirus, meningococcal and malaria vaccines in developing countries. To optimize the usefulness of vaccine economics research, conceptual issues, such as how to set values for the prevention of illness and how to maximize social equity through investments in vaccines, must be addressed. PMID- 12901583 TI - Stress-associated immunomodulation and its implications for responses to vaccination. AB - The rapidly growing field of psychoneuroimmunology involves the elucidation of the complex interactions between the CNS, the endocrine system and the immune system and its effects on health. Although the mechanisms involved in this bidirectional communication is not yet fully understood, studies in psychoneuroimmunology have shown that stress, through the hypothalamic-pituitary adrenal and the sympathetic-adrenal-medullary axes, can induce modulation of the immune system. In this review, we discuss human studies and animal models, which focus on psychological stress and its effects on the immune response to vaccination, emphasizing the implications of these effects on health. PMID- 12901584 TI - Hepatitis B vaccine and risk of multiple sclerosis. AB - The possibility that hepatitis B vaccine may cause or exacerbate multiple sclerosis stems from several case reports of onset or recurrence of symptoms of CNS demyelination shortly following vaccination. It is difficult, however, to infer causation from individual case reports since they may simply represent coincidental temporal associations with vaccination. There is only weak, nonspecific evidence to support the biological plausibility of an association between hepatitis B vaccine and multiple sclerosis. Epidemiological studies have found that hepatitis B vaccine does not increase the risk of developing multiple sclerosis or cause exacerbations. The US Institute of Medicine and other review panels have concluded that the evidence favors rejection of a causal association between hepatitis B vaccine and multiple sclerosis. PMID- 12901585 TI - Epstein-Barr virus vaccine: a cytotoxic T-cell-based approach. AB - Epstein-Barr virus infects more than 95% of the human population and is linked to infectious mononucleosis as well as a series of geographically-defined cancers. To date, there is no prophylactic or therapeutic vaccine available for Epstein Barr virus-associated diseases. New immunotherapeutic approaches, based on cytotoxic T-cells, are being developed depending on the degree of Epstein-Barr virus antigen expression in infected cells. It is hoped that these approaches will provide enough impetus for cytotoxic T-cell-based vaccine development. Approaches for developing vaccines towards the different Epstein-Barr virus associated diseases are discussed. PMID- 12901586 TI - Melioidosis vaccines. AB - Melioidosis is a disease caused by the facultative intracellular pathogen Burkholderia pseudomallei iand is associated with a high mortality rate. Melioidosis is endemic in the tropics of southeast Asia and northern Australia and is of worldwide concern, particularly as it is a potential agent of bioterrorism or biological warfare. Also of concern is the lack of a fully effective antibiotic regime, as cases of bacteremia have unacceptably high mortality rates and relapse of melioidosis is common. This review focuses on the approaches that have been undertaken towards the development of an effective vaccine against this disease and highlights current strategies being used to move towards finalizing such a vaccine. PMID- 12901587 TI - Current progress in enteropathogenic and enterohemorrhagic Escherichia coli vaccines. AB - Enteropathogenic and enterohemorrhagic Escherichia coli are important causal agents of infectious diarrhea, particularly amongst pediatric populations. While enteropathogenic E. coli is a significant health threat in developing countries, enterohemorrhagic E. coli causes sporadic, sometimes deadly outbreaks of hemorrhagic colitis, with a serious complication, hemolytic uremic syndrome, ocurring in a proportion of cases. This review discusses the pathogenesis of enterohemorrhagic and enteropathogenic E. coli, the host immune response and the current application of this knowledge towards efficacious vaccine strategies. Several lines of investigation indicate the feasibility of such strategies and justify further development of a vaccine targeting these significant intestinal pathogens. PMID- 12901588 TI - Live bacteria as the basis for immunotherapies against cancer. AB - For more than a century, bacteria and bacterial products have been used for the treatment of cancer. Starting from the practical observation of tumor regression in individuals with concomitant bacterial infection, the field has evolved into some standard clinical practices, such as the use of BCG for the treatment of superficial bladder cancer. However, in the last few years, new applications have started to emerge that may profoundly change the perspective of the field. BCG can be engineered to express cytokines to improve its efficacy. Bacteria such as Salmonella and Listeria can be attenuated by genetically-defined mutations and provide effective vehicles for DNA vaccines encoding tumor-associated antigens. Salmonella and nonpathogenic strains of Clostridium can selectively accumulate in tumors in vivo, providing attractive delivery systems to target immunomodulatory molecules and therapeutic agents to the tumor site. Many of these new developments have been attempted for prophylactic or therapeutic vaccination in several different experimental models of cancer and in many cases, results from clinical trials are now emerging. There is still some way to go before achieving products that could be in routine use, but the field has great promise for the development of more effective immunotherapies for several different cancers. In this paper, we will review the current state of such applications and highlight some of the directions that the field may take. PMID- 12901589 TI - RNA-transfected dendritic cells. AB - Based on their unique ability to stimulate primary immune responses, dendritic cells are the most potent antigen-presenting cells known. This ability stems from the fact that they are very efficient at the uptake and processing of antigen and they express high levels of major histocompatibility complex class I and class II, as well as costimulatory molecules, which are required to prime naive cytotoxic T-cells. Many groups of investigators have tried to take advantage of these features by developing dendritic cell-based vaccines against tumors and infectious diseases. While the basic principle in these studies is the same- dendritic cells pulsed with antigen are used to elicit cytotoxic T-cell responses -the methods used are varied. This is particularly true with respect to the nature of the antigen used and the method of antigen delivery. In this article, we will focus on the use of RNA as a form of antigen with which to load dendritic cells. We will discuss the rationale behind using RNA as an antigen source and will review recent studies in both murine and human settings that use RNA-pulsed dendritic cells as vaccines. PMID- 12901591 TI - Therapeutic AIDS vaccine looks promising. PMID- 12901590 TI - The power of 'box a'. PMID- 12901592 TI - Immunogenicity of Twinrix in older adults: a critical analysis. AB - Twinrix (GlaxoSmithKline Biologicals, Rixensart, Belgium) is the first combined vaccine to provide protection against both hepatitis A and B. This review presents a critical analysis of antibody responses stratified by age following vaccination with Twinrix in 264 adults aged above 40 years. A month after completion of a 0-, 1-, 6-month vaccination schedule with Twinrix, a good response was observed for both anti-HAV and anti-HBs serum antibodies, suggesting that is an effective vaccine in older adults. PMID- 12901593 TI - Designing pediatric vaccine formularies and pricing pediatric combination vaccines using operations research models and algorithms. AB - The National Immunization Program, housed within the Centers for Disease Control and Prevention in the USA, has identified several challenges that must be faced in childhood immunization programs to deliver and procure vaccines that immunize children from the plethora of childhood diseases. The biomedical issues cited include how drug manufacturers can combine and formulate vaccines, how such vaccines are scheduled and administered and how economically sound vaccine procurement can be achieved. This review discusses how operations research models can be used to address the economics of pediatric vaccine formulary design and pricing, as well as how such models can be used to address a new set of pediatric formulary problems that will surface with the introduction of pediatric combination vaccines into the US pediatric immunization market. PMID- 12901594 TI - Vaccine safety surveillance using large linked databases: opportunities, hazards and proposed guidelines. AB - Combined administrative databases are referred to as 'large linked databases' because of their relatively large size and the need for linkage of different data sets that were created separately from each other. Such linked databases have become popular in vaccine safety surveillance. Whereas their use offers some unique opportunities, their increasingly widespread use can also lead to wrongful linkage of vaccines to adverse events. We review the opportunities and hazards of using large linked databases for vaccine safety surveillance and propose some guidelines to increase the reliability of the outcomes. We also offer our opinion on the future use of large linked databases for vaccine safety surveillance purposes. PMID- 12901595 TI - Advances in the development of bacterial vector technology. AB - The demand for new and improved vaccines against human diseases has continued unabated over the past century. While the need continues for traditional vaccines in areas such as infectious diseases, there is an increasing demand for new therapies in nontraditional areas, such as cancer treatment, bioterrorism and food safety. Prompted by these changes, there has been a renewed interest in the application and development of live, attenuated bacteria expressing foreign antigens as vaccines. The application of bacterial vector vaccines to human maladies has been studied most extensively in attenuted strains of Salmonella. Live, attenuated strains of Shigella, Listeria monocytogenes, Mycobacterium bovis BCG and Vibrio cholerae provide unique alternatives in terms of antigen delivery and immune presentation, however and also show promise as potentially useful bacterial vectors. PMID- 12901596 TI - Bacterial ghosts as carrier and targeting systems for mucosal antigen delivery. AB - The application of new strategies to develop effective vaccines is essential in modern medicine. The bacterial ghost system is a novel vaccine delivery system endowed with intrinsic adjuvant properties. Bacterial ghosts are nonliving gram negative bacterial cell envelopes devoid of cytoplasmic contents while maintaining their cellular morphology and native surface antigenic structures including bioadhesive properties. They are produced by PhiX174 protein E-mediated lysis of gram-negative bacteria. The intrinsic adjuvant properties of bacterial ghost preparations enhance immune responses against envelope-bound antigens, including T-cell activation and mucosal immunity. Since native and foreign antigens can be expressed in the envelope complex of ghosts before E-mediated lysis, multiple antigens of various origin can be presented to the immune system simultaneously. In addition, the extended bacterial ghost system represents a platform technology for specific targeting of DNA-encoded antigens to primary antigen-presenting cells. The potency, safety and relatively low production cost of bacterial ghosts offer a significant technical advantage, especially when used as combination vaccines. PMID- 12901597 TI - Antibody therapy for Alzheimer's disease. AB - The economic, social and emotional impact of Alzheimer's dementia is increasing dramatically as greater numbers live to advanced ages. The dearth of effective therapies has led to innovative approaches to treat the disease. This review summarizes the rationale, progress and setbacks regarding the use of antibody based therapies to treat Alzheimer's disease and discusses future directions for this approach in Alzheimer's and other disorders. PMID- 12901599 TI - Genetic engineering of pollen allergens for hayfever immunotherapy. AB - Type I allergy is an immunoglobulin E-mediated disease, representing a major health problem affecting more than 25% of the world's population. Grass, birch and ragweed pollen are a major source of environmental allergen. Specific immunotherapy is clinically proven to be an effective treatment for allergic conditions that involve the administration of crude extracts prepared from natural sources with potential life-threatening anaphylactic side effects. Recent developments in the molecular biology of pollen allergens have made it possible to design novel therapeutic approaches for improved and safer forms of specific immunotherapy. Hypoallergenic forms of major allergens with reduced immunoglobulin E epitopes have been produced using genetic engineering, whilst preserving other characteristics of the molecule that are able to induce a protective response. These modified forms are expected to make allergen-specific immunotherapy more widely used. PMID- 12901600 TI - Immunological mechanisms of specific immunotherapy with pollen vaccines: implications for diagnostics and the development of improved vaccination strategies. AB - This review deals with specific immunotherapy performed with pollen vaccines. Pollen allergens, the immunological mechanism of allergic inflammation and the mechanisms behind successful specific immunotherapy treatment of seasonal pollen allergy shall be discussed. The different approaches to develop improved regimes for specific immunotherapy based on new diagnostic tools, improved pollen vaccines and alternative routes for administration of the vaccines will be highlighted. Diagnostic tools to select patients for specific immunotherapy and for monitoring the treatment are quantitative measurements of allergen-specific immunoglobulin E, G and G4 antibodies and component-resolved diagnostics based on immunoglobulin E binding to individual allergenic components. Future pollen vaccines may consist of hypoallergenic recombinant allergens and T-helper cell type 1-inducing immunostimulatory adjuvants. Both sublingual and nasal administration of pollen vaccines for specific immunotherapy may be used more in future. PMID- 12901598 TI - Rational design of peptide vaccines for autoimmune disease: harnessing molecular recognition to fix a broken network. AB - Autoreactive T-cells and antibodies are found at low levels in normal individuals and are thought to be kept at bay by regulatory T-cells and a network of idiotypic and anti-idiotype-bearing antigen receptors on lymphocytes as well as idiotypic anti-idiotypic antibodies. Disruption of this network by genetic, environmental and unknown factors is thought to result in autoimmune diseases. An obvious, ideal and specific therapy for such disorders would be to harness this regulatory network to re-establish immunologic homeostasis. In practice, however, this is not an easy task as most autoimmune diseases involve polyclonal responses to self antigen. Thus, we are faced with the conundrum of not knowing which autoreactive idiotype-bearing antibody or antigen receptor(s) to target in order to restore or induce network regulatory function. The thesis of this review is that understanding a fundamental property governing peptide/protein shape can be used in part to circumvent the problems of self reactivity and polyclonality in autoimmune disorders. More specifically, an algorithm has been developed to design peptide vaccines with shapes that are thought to be complementary in contour to self epitopes which seem to be the focus of autoimmunity. In theory, such complementary shapes should be engendered in certain autoreactive antigen receptors--these complementary constructs consequently represent receptor mimetics. By targeting an immune response against such mimetics, one generates a polyclonal anti-idiotype response that matches the complexity of the autoimmune response itself. This article will describe the algorithm for vaccine design, summarize the in vitro and in vivo evidence for its efficacy and discuss possible therapeutic utility in human autoimmune diseases. PMID- 12901601 TI - Polio vaccines and the cessation of vaccination. AB - Poliomyelitis is on the verge of eradication as a result of heroic immunization efforts by a consortium including the World Health Organization, Centers for Disease Control and Prevention, United Nations International Children's Emergency Fund and Rotary International. The live vaccine that is the major tool of the program evolves rapidly in vaccine recipients, can spread to their contacts to some extent and can on rare occasions cause poliomyeltis. Some of the difficulties that this implies for the eventual complete cessation of vaccination are proving to be real not theoretical. PMID- 12901602 TI - Tropical disease vaccines. AB - Infectious diseases remain a key cause of morbidity and mortality in the Tropics. Many of these diseases are already preventable by effective vaccines. The Extended Program of Immunization provides protection against six serious infections. However, this program is still not optimally implemented in all developing countries. The Extended Program of Immunization could benefit from the addition of several vaccines, but unfortunately the cost is prohibitive in many areas of the Tropics. For some of the most important infections, such as malaria and HIV, vaccines are not yet available. Novel strategies for vaccine development and policy implementation offer the best hope to combat the infections targeted by the World Health Organization as causing the highest annual death toll worldwide. PMID- 12901603 TI - Pertussis: persistent pathogen, imperfect vaccines. AB - Routine use of pertussis vaccines has diminished the incidence of this disease but has not eliminated the pathogen. Pertussis remains a significant cause of disease in both very young infants and in the adolescent and adult populations. Acellular pertussis vaccines have fewer adverse reactions compared with whole cell pertussis vaccines. Although efficacious against severe disease, current vaccines may not be as efficacious against milder forms of infection. New methodologies for understanding disease pathogenesis, immune responses and vaccine development are needed to effectively interrupt continued transmission of this pathogen. PMID- 12901605 TI - Current research and future prospects in the development of vaccines against gastrointestinal nematodes in cattle. AB - Vaccination is being considered as the most feasible alternative for anthelmintic drugs to control gastrointestinal nematode infections in cattle. However, despite the identification of several candidate protective antigens, no vaccines against gastrointestinal nematode parasites are currently available. The main problems that hamper the development of nematode vaccines in ruminants are that vaccination with recombinant nematode proteins produced in bacterial or eukaryotic expression systems did not induce a protective immune response and no suitable antigen delivery system is available for presentation of protective worm antigens to the bovine mucosal immune system. The present review will focus on recent advances and remaining obstacles in vaccine development against gastrointestinal nematodes in cattle, in particular against the abomasal parasite Ostertagia ostertagi. PMID- 12901604 TI - Contemporary approaches to designing and evaluating vaccines against Chlamydia. AB - The clinically relevant pathologic consequences of primary ocular, genital, or respiratory human infection by members of the genus Chlamydia are conjunctivitis, cervicitis, urethritis and sinusitis. The major complications and sometimes debilitating evolutionary outcomes of these infections include: trichiasis and cicatrizing trachoma, endometritis or pelvic inflammatory disease and involuntary tubal factor infertility and bronchopulmonary pneumonia. These diseases, in addition to other chlamydia-associated chronic syndromes (e.g., artherosclerosis, multiple sclerosis and Alzheimer's disease), pose serious public healthcare and huge budgetary concerns. The current medical opinion is that an efficacious prophylactic vaccine is a sine qua non--to control the morbidity of chiamydial infection in the human population. The research goal for an efficacious human chlamydial vaccine has faced key challenges to define the elements of protective immunity to facilitate vaccine evaluation, the judicious selection of appropriate vaccine candidates that possess stable antigenic and immunologic properties and the development of effective delivery vehicles and adjuvants to boost immune effectors to achieve long-term protective immunity. Progress in the functional immunobiology of Chlamydia has established the essential immunologic paradigms for vaccine selection and evaluation, including the obligatory requirement for a vaccine to induce T-helper Type 1 immune response that controls chlamydiae. Recent advances in chlamydial genomics and proteomics should enhance the identification of likely chlamydial gene products that fulfill the antigenic requirements of putative vaccine candidates. Major inroads are however needed in the construction and development of novel and effective delivery systems, such as vectors and adjuvants. This review summarizes the status of contemporary chlamydial vaccine research and promising trends fueling the growing optimism for an efficacious vaccine. The unified approach to vaccines for the genus Chlamydia is validated by the several conserved genes and common immunogenic proteins among member species and the similarity of immune effectors controlling Chlamydia species in animals and humans. PMID- 12901606 TI - Angiotensin II-acetylcholine noncovalent complexes analyzed with MALDI-ion mobility-TOF MS. AB - Matrix-assisted laser desorption ionization-ion mobility-orthogonal time-of flight mass spectrometry (MALDI-IM oTOF MS) is a new technique that allows laser desorbed ion to be preseparated on the basis of their shape prior to mas analysis. Using this instrument, we tested the postulate that addition of a quaternary ammonium compound such as acetylcholine to the model phosphorylated peptide angio tensin II would enhance its detection by MALDI in two ways. First of all, the acetylcholine-peptide complex could ionize more efficiently than the bare phosphopeptide. Furthermore the ion mobility could separate the complex ion on the basis of its charge/volume from isobaric interferences, which would otherwise limit detection sensitivity. PMID- 12901607 TI - Simple tests to detect errors in high-throughput genotype data in the molecular laboratory. AB - With the advent of high-density DNA marker data sets for the mouse and other model systems, 100 or more genotype are routinely generated from large groups of mice. Issues of the accuracy and reliability of the genotyping are extremely important but often not addressed until genetic analysis is conducted. Simple tests that rely on the robust predictions arising from Mendelian genetics can be made quickly in the molecular laboratory as the data are generated, and require only a spreadsheet program. In this report, genotype data from 392 mice tested at 96 marker sites were analyzed for errors that are typical when handling large volumes of data generated in a repetitive process. The testing consisted of: (1) repeating the genotyping of approximately 1% of the samples; (2) examining the deviation from the expected segregation ratio ( 1:2:1 ) on a marker-by-marker basis; and (3) testing the correlation of the genotype at one marker with that at neighboring genetic markers on a chromosome. These three steps allowed analysis at the level of the microtiter plate, where errors are most likely to occur. A set of 96 dinucleotide repeat markers that are polymorphic between the C57BL/6J and DBA/2J mouse strains and can be multiplexed is reported for use in other genotyping projects. PMID- 12901608 TI - A rapid method for manual or automated purification of fluorescently labeled nucleic acids for sequencing, genotyping, and microarrays. AB - Fluorescent dyes provide specific, sensitive, and multiplexed detection of nucleic acids. To maximize sensitivity, fluorescently labeled reaction products (e.g., cycle sequencing or primer extension products) must be purified away from residual dye-labeled precursors. Successful high-throughput analyses require that this purification be reliable, rapid, and amenable to automation. Common methods for purifying reaction products involve several steps and require processes that are not easily automated. Prolinx, Inc. has devel oped RapXtract superparamagnetic separation technology affording rapid and easy-to-perform methods that yield high-quality product and are easily automated. The technology uses superparamagnetic particles that specifically remove unincorporated dye labeled precursors. These particles are efficiently pelleted in the presence of a magnetic field, making them ideal for purification because of the rapid separations that they allow. RapXtract-purified sequencing reactions yield data with good signal and high Phred quality scores, and they work with various sequencing dye chemistries, including BigDye and near-infrared fluorescence IRDyes. RapXtract technology can also be used to purify dye primer sequencing reactions, primer extension reactions for genotyping analysis, and nucleic acid labeling reactions for microarray hybridization. The ease of use and versatility of RapXtract technology makes it a good choice for manual or automated purification of fluorescently labeled nucleic acids. PMID- 12901610 TI - Peripheral tissue distribution of orphanin FQ precusor mRNA in stroke-prone spontaneously hypertensive rats. AB - The heptadecapeptide orphanin FQ (OFQ) is a recently discovered neuropeptide that exhibits structural features reminiscent of the opioid peptides and that is an endogenous ligant to a G protein-coupled receptor sequentially related to the opioid receptors. OFQ was originally isolated from brain, but the presence of OFQ in peripheral tissues, especially in cardiovascular system, has not been clarified. The present study was designed to investigate the peripheral tissue distribution of OFQ precusor mRNA in stroke-prone spontaneously hypertensive rats (SHRSP) and compare the difference of OFQ precusor mRNA expression in aorta or cultured vascular smooth muscle cells (VSMCs) between SHRSP and wistar-Kyoto normotensive (WKY) rats. By using quantitative reverse transcription-polymerase chain reaction (RT-PCR), OFQ precusor mRNA was detected in aorta and ovary at high levels comparable with the amounts found in brain. Moderate expression was found in testis, while a little OFQ precusor mRNA could be detected in atrium. All other peripheral tissues examined from SHRSP, including ventricle, liver, lung and kidney, showed no expression of OFQ precusor mRNA. In the vascular system, OFQ precusor mRNA was expressed in aorta, pulmonary artery, renal artery and vein at high levels comparable with the amounts found in brain. We also found that OFQ precusor mRNA levels were much higher in aorta or cultured VSMCs from SHRSP than those from WKY rats. In conclusion, the present study has shown that OFQ precusor mRNA is present in some peripheral tissues, especially in cardiovascular and reproductive system, suggesting that OFQ possibly involves in the regulation of cardiovascular and reproductive functions. PMID- 12901609 TI - The use of real-time reverse transcriptase PCR for the quantification of cytokine gene expression. AB - Real-time reverse transcriptase polymerase chain reaction (RT-PCR) is becoming a widely used method to quantify cytokines from cells, tissues, or tissue biopsies. The method allows for the direct detection of PCR product during the exponential phase of the reaction, combining amplification and detection in a single step. Using TaqMan chemistry (Applied Biosystems, Foster City, CA) and the ABI Prism 7700 Sequence Detection System (Applied Biosystems), we validated a large panel of murine and human cytokines, as we as other factors playing a role in the immune system, such a chemokines and apoptotic markers. Although the method allows fast, sensitive, and accurate quantification, different control assays are necessary for the method to be reliable. By construction of complementary DNA (cDNA) plasmid clones, standard curves are generated that allow direct quantification of every unknown sample. Furthermore, the choice of a reliable housekeeping gene is very important. Finally, co-amplification of contaminating genomic DNA is avoided by designing sets of primers located in different exons or or intron-exon junctions. In conclusion, the real-time RT-PCF technique is very accurate and sensitive, allows high through put, and can be performed on very small samples. The development of real-time RT-PCR has resulted in an exponential increase in its use over the last couple of years, and the method has undoubtedly become the standard for quantifying cytokine patterns, clarifying many functional properties of immune cells and their associated diseases. PMID- 12901611 TI - The relationship of HLA-DRB1 * 0701 allele with the psoriasis vulgaris. PMID- 12901612 TI - The study on related genes in the neoplastic transformation of immortalized human fetal tracheal fibroblast cells induced by irradiation. AB - In this study, we investigated the genes related to the transformation of immortalized human fetal tracheal fibroblast cell line induced by alpha particles by means of differential display mRNA method. The result revealed that there were 23 DNA fragments that were expressed intensively in alphaSHTF cells (SHTF cells forming clone on agar after irradiated by alpha particles emitted by 238Pu) only and not in SHTF (SV40-immortalized human fetal tracheal fibroblast) cells. Northern dot confirmed two fragments, C17-5, C23-1 which showed intensive mRNA expression in alphaSHTF cells, but not in SHTF cells. The length of the C17-5 fragment was 310bp. Searching in BLAST database revealed that the C17-5 fragment might be an unknown sequence. PMID- 12901613 TI - Effects of musical meditation training on auditory mismatch negativity and P300 in normal children. AB - The auditory mismatch negativity (MMN) and P300 of event-related potentials were compared in normal children either with or without musical meditation training. The experimental group consisted of 11 subjects who had been trained with musical meditation for six months and the control group consisted of 12 subjects (matched for age, sex and grade) who had not received musical meditation. MMN amplitudes in the trained children were larger than those in the control group. In addition, the MMN amplitudes were identical in attend and ignore conditions for both groups. This evidence suggests that auditory brain function has been affected by musical meditation training. It thus suggests that the MMN is capable of assessing changes to the brain function in normal subjects. There were no significant differences in the P300 latencies and amplitudes between the two groups. This result suggests that MMN and P300 may reflect different aspects of the brain function. PMID- 12901614 TI - Experimental study on preservation of rat fatty liver. AB - OBJECTIVE: To investigate the effects of cold preservation on rat fatty liver. METHODS: We observed the changes of portal perfusion pressure, endothelin-1, enzymes release in the effluent and mortality of sinusoid lining cell after 0h, 6h, or 12h preservation respectively and a subsequent 30 min reperfusion in rat fatty liver groups and control groups by using isolated perfused rat liver model. And we compared fatty liver groups with control groups by these indices. RESULTS: There was no obvious difference between mildly fatty liver group and control group after long time (12h) preservation, between moderately fatty liver group and control group after short time (6h) preservation, between severely fatty liver group and control group without preservation (0h), while preservation reperfusion injury was more severe in moderately fatty liver group than in control group after long time (12h) preservation and in severely fatty liver group than in control group after short time (6h) preservation. CONCLUSIONS: The authors suggested that a mildly fatty liver donor could be used in the same way as nonfatty liver and a moderately fatty liver donor could be used depending on the time of preservation and the balance of the emergent needs of recipient and donor organ supply, while severely fatty liver donor should be discarded without hesitation. PMID- 12901615 TI - Transesophageal echocardiographic monitoring during radiofrequency ablation of tachycardia. PMID- 12901616 TI - Evolutionary relationships among seventeen human papillomavirus genotypes. AB - The DNA and the protein sequences of the six major open reading frames (ORFs) of 17 human papillomavirus (HPV) genotypes have been carefully aligned. Conserved primary and probable secondary structural features of the proteins have been identified. From the analysis of variations in the aligned positions, the most consistent evolutionary relationships have been reconstructed for each gene and for the genomes as a whole. The L1 gene encoding the major capsid protein and the E1 gene encoding a replication protein are the most conserved. The hinge region in E2 is the most divergent. With varying degrees of sequence relatedness, the major groups and subgroups of HPV genotypes can be recognized, these clusters: the mucosotrophic virus types (33, 58), (16, (31, 35)), (18, 39), (51), (6b, 11); mucosal/cutaneous transitional virus types (2a, 57); and the cutaneous virus types (47, (5, 8)), (1a), (41), closely reflect epithelial tissue-type predilections and clinical pathogenicities. PMID- 12901617 TI - Over-expression of the RAB5 gene in human lung adenocarcinoma cells with high metastatic potential. AB - The objective of this study is to better understand the molecular mechanism of tumor invasion and metastasis, and isolating tumor metastasis-related genes. Two human lung adenocarcinoma cell lines AGZY-83a and Anip973 were studied. Anip973 was derived from AGZY-83a, but it manifested a very much higher metastatic potential than the parent line. Differential cDNA fragments were isolated by using the techniques of mRNA differential display, and analyzed by means of molecular cloning and sequencing. The expression of RAB5A gene in clinical samples of non-small cell lung cancer was determined by RT-PCR. There were significant differences between AGZY-83a and Anip973 in gene expression. Part of the differential cDNA fragments were cloned and sequenced. We found that there was over-expression of RAB5A gene in the Anip973 cell line. And there was over expression of RAB5A gene in those samples of clinical lung cancer showing metastasis. In conclusion, the expression or over-expression of RAB5A gene was associated with the metastatic phenotype of Anip973. Probably, over-expression of RAB5A gene in non-small lung cancer may serve as a diagnostic marker for metastasis. PMID- 12901618 TI - The relationship between expressions of N-myc and c-myc oncogenes in neuroblastoma: an in situ hybridization and immunocytochemical study. AB - N-myc gene amplification is the most characteristic feature of neuroblastoma. c myc oncogene, another member of myc gene family, plays an important role in cell proliferation and differentiation. Both of them may contribute to tumorigenesis of neuroblastoma. In this study we use the in situ hybridization and immunocytochemical methods to test the frequencies of N-myc and c-myc expressions in 20 cases of human neuroblastoma at mRNA and protein levels. The positive rates of the expression of N-myc are 90% and 100% detected by in situ hybridization and immunocytochemical methods respectively. The positive rates of c-myc are 80% and 85% respectively. Sixty percent of the 20 specimens tested by in situ hybridization and 55% by immunocytochemistry show an inverse relationship between the expressions of these two oncogenes and this may indicate that there are different gene expression controlling mechanisms in different cases. PMID- 12901619 TI - Preparation of recombinant alpha2 antigen of M. leprae in E. coli and the application for sero-diagnosis of leprosy. PMID- 12901620 TI - Study of loss of heterozygosity at DCC and APC/MCC genetic loci of gastric cancer. AB - In order to assess the role of the changes of DCC and APC/MCC genes in the development and progression of gastric cancer, the loss of heterozygosity (LOH) of these genetic loci was investigated in 45 surgical specimens of gastric cancer with PCR-RFLP. The rate of LOH was 30.0% (9/30) at APC/MCC gene and 33.3% (15/45) at DCC gene. LOH was found in both intestinal and gastric types of gastric cancer and the rate of LOH of DCC gene was significantly higher in stages III to approximately IV gastric cancer (48.0%) than in stages I to approximately II (15.0%) (P<0.05). LOH of APC/MCC gene could be found in both early and advanced stages of gastric cancer. These findings suggest that changes of DCC and APC/MCC genes are involved in the development and progression of the intestinal and gastric types of gastric cancer. PMID- 12901621 TI - Alteration in enterocyte gene expression may explain structural and functional changes following glutamine supplemented parenteral nutrition. AB - Following extensive bowel resection, the intestinal tract undergoes a variety of adaptive responses to enhance bowel function. The purpose of this study was to determine the effect of glutamine-supplemented parenteral nutrition on mucosal cellularity and gut function. In addition, enterocyte gene expression of two relevant systems was also characterized and related to the structural and functional changes that occurred. Male Wistar rats underwent a 60% small bowel resection and jugular vein catheterization and were randomized into two groups. The control group (n = 10) received a standard intravenous nutritional solution and the study group (n = 10) received a similar solution but enriched with alanylglutamine dipeptide. After 7 days blood was taken for amino acid analysis, and bowel was harvested to determine mucosal morphology and expression of mucosal cell glutaminase and IGF-I mRNA. Mesentery lymphnodes were cultured to determine the presence of bacteria and thus access bacteria translocation. Serum glutamine concentration and mucosal architecture were maintained in the study group compared to the controls. Seventy percent of lymphnodes were cultured positive in control vs. only 20% in the study group (P < 0.05). Jejunal mucosal glutaminase and ileum mucosal IGF-I mRNA increased twofold and threefold respectively compared to control animals. Parenteral nutrition supplemented with alanyl glutamine dipeptide supports mucosal cellularity and regional immune function in rodents following intestinal resection, These alterations are associated with enhanced enterocyte expression of glutaminase and IGF-I. These changes may facilitate the structural and functional alterations which were observed in the glutamine treated animals. PMID- 12901622 TI - Comparison of bone mass in forearm, lumbar vertebra and hip by single and/or dual energy X-ray absorptiometry. AB - OBJECTIVE: Bone mineral density (BMD) was measured in normal Chinese women with single X-ray absorptiometry (SXA) and dual energy X-ray absorptiometry (DEXA). These two methods were compared to evaluate the sensitivity in reflecting bone loss. METHODS: Measurements were performed in 300 women aged 20 to approximately 79 (5 for each age). The 8mm distal, 1/4 distal, ultra distal site of the nondominant forearm were measured with SXA; the AP spine (L2-4), right femoral neck (Neck), Ward's triangle (Ward) and trochantor (Troch) were measured with DEXA. Ten women had 5 repeated measurements to evaluate the reproducibility of the equipments. RESULTS: The BMD peak for Neck and Ward was found between age 20 to approximately 29, for ultra distal and L2-4 at age 30 to approximately 39, for 8mm distal, 1/4 distal and Troch at age 40 to approximately 49. After reaching the peak values, BMD at all sites decreased with increasing age. The highest rate of yearly loss of BMD was at 8mm distal. Results of measurement showed significant correlation between SXA and DEXA. Comparing with premenopausal women, the bone loss rate in postmenopausal women was faster and highest up to 11 to approximately 15 years and the highest rate was at the Ward. CONCLUSION: The present study gives the normal values of BMD at seven sites for normal Chinese women. The ages of BMD peak were different at seven sites. The yearly loss of BMD was highest at 8mm distal, it seemed to be the sensitive region for bone loss with age. The postmenopausal women had highly significantly loss bone mass than the premenopausal women. Comparing with premenopause, postmenopause BMD for Ward's triangle with DEXA seemed to be most sensitive to the effect of menopause. PMID- 12901623 TI - Suppressive effect of carotenoids on the luminol dependent chemiluminescence of the stimulated rat macrophages. AB - Rat macrophages stimulated by phorbol 12-myristate 13-acetate (PMA) produced significant chemiluminescence in luminol. Carotenoids suppressed the luminol dependent chemiluminescence at the beginning and after two minutes of the stimulation. Canthaxanthin and bixin had higher suppressive activity that beta carotene and lutein. The measurement of the absorption spectra of carotenoids showed that the absorption by carotenoids was diminished during the stimulation of macrophages. The results suggest that the suppressive effect of carotenoids may be due to their scavenging the reactive oxygen metabolites released by macrophages. PMID- 12901625 TI - A dynamic epidemiological study of pathogenic fungi in China. PMID- 12901624 TI - Coral as a carrier for recombinant human bone morphogenetic protein-2. AB - By combining coral with recombinant human bone morphogenetic protein-2 (rhBMP-2), rhBMP-2/coral composite was obtained in this study. Following implantation of the composite into the muscle pouches of mice, cartilage growth was induced in the pores or on the surface of the implants at one week, woven bone at three week and lamellar bone with bone marrow at six week, and coral was absorbed partially. The induced formation of endochondral bone was time-related and rhBMP-2 dose-related. The results of this study indicate that the composite possesses a superior ability of osteogenesis, and coral acts as one of the most suitable rhBMP-2 slowrelease carriers currently available. The composite will be a new type of bone substitute to be used in orthopaedics and maxillofacial surgery. PMID- 12901626 TI - MR imaging of coronary artery bypass graft: a preliminary study. AB - OBJECTIVE: To make a preliminary investigation of the patency and function of coronary artery bypass grafts (CABG) by magnetic resonance (MR) images and to establish a suitable method for follow-up study after CABG operation among Chinese. METHODS: MR imaging was performed with a Toshiba 1.5-T unit in 27 patients with 74 grafts. All patients were examined with a breath-hold ECG-gated two-dimensional fast field echo (FFE) sequence to evaluate the patency of bypass grafts, among them 16 patients with 42 grafts were further examined with a phase shift magnetic resonance angiography flow (PSMRAflow) sequence to evaluate the grafts patency as well as the flow velocity and flow volume vs. time. RESULTS: The results showed that 66 of the 74 grafts in the patients of the present series studied with FFE were patent with a patency rate of 89.2%. The results evaluated both with FFE and PSMRAflow remained the same except that two grafts were patent with FFE and the results with PSMRAflow were uncertain. Diastolic perfusion pattern curves were found in 25 of the 32 grafts in patients of the present series. Comparing the flow curves of the grafted left internal mammary artery with those of the native right internal mammary artery in 7 patients, the systolic peak velocity value (SPV) of the grafted arteries was significantly lower than that of the ungrafted ones, whereas the diastolic peak velocity value (DPV) and the ratio of DPV to SPV were significantly greater than that of the ungrafted ones. CONCLUSION: The FFE and PSMRAflow sequences were efficient in evaluating patency and obtaining the curves of flow velocity and volume of the bypass grafts. Therefore, they may offer a non-invasive screening method for follow-up study in patients after CABG surgery, although its accuracy should be further evaluated in more patients and comparatively studied with other methods. PMID- 12901627 TI - C-type natriuretic peptide inhibits upregulation of alpha1-adrenoceptor and inositol 1,4,5-trisphosphate receptor in rat vascular smooth muscle after vascular endothelial injury. AB - OBJECTIVE: In a model of balloon injury of rat aortic endothelium, the effects of C-type natriuretic peptide (CNP) on alpha1-adrenoreceptor and inositol 1,4,5 triphosphate (IP3) receptor were studied. METHODS: Aortic injuries were produced by vascular endothelium-denudation. Alpha1-adrenoreceptor in smooth muscle sarcolemma and IP3 receptor in smooth muscle sarcoplasmic reticulum in the rat aorta were assayed by radioactive analysis method. RESULTS: It was found that neointima was formed and the contents of DNA, collagen and elastin of each intima media were sigificantly increased in 7 days and 21 days after balloon injury of rat aorta. Alpha1-adrenoreceptor in smooth muscle sarcolemma and IF3 receptor in sarcoplasmic reticulum were also upregulated. Results also showed that the administration of CNP i.p. significantly decreased the contents of DNA, collagen and elastin of each intima-media, and inhibited the up-regulation of alpha1 adrenoreceptor and IP3 receptor. CONCLUSION: The inhibition of the up-regulation of alpha1-adrenoreceptor and IP3 receptor by CNP might be one of the mechanisms of its suppressive action on intimal proliferation. PMID- 12901628 TI - The relationship between the N-glycosylation of acetylglucosaminyltransferase V and its activity. AB - OBJECTIVE: The goal of this paper is to investigate the relationship between the N-glycosylation of acetylglucosaminyltransferase V (GlcNAcT-V) and its activity and to know which site among the 6 N-glycosylation sites in the GlcNAcT-V gene is the most important. METHODS: Wild type of GlcNAcT-V was transfected into COS-7 cells and its activity was measured 48 h later. The first site (Asn 110) was mutated with site-directed mutagenesis and transfected into COS-7 cells. RESULTS: It was found that after the cells were added tunicamycin (TM, 1 microg/ml), the activity was 11.7% of the wild type. The activity of the cells with mutating GlcNAcT-V was about 12.0% of the wild type. RT-PCR showed that there was no significant change in mRNA expression among the three groups. CONCLUSION: The N glycosylation is important for its activity. Our results suggest that the N linked carbohydrates on GlcNAcT-V are required for the posttranscriptional activity of the enzyme. PMID- 12901629 TI - Effects of Radix Angelicae sinensis and shuanghuanglian on a rat model of chronic Pseudomonas aeruginosa pneumonia. AB - OBJECTIVE: To study the effect of two kinds of Chinese herbal medicine, Radix angelicae sinensis (RAS) ([Chinese characters: see text]) and Shuanghuanglian (SHL) ([Chinese characters: see text]) on chronic Pseudomonas aeruginosa (PA) lung infection in a rat model mimicking cystic fibrosis (CF). METHODS: Rats were divided into RAS, SHL and control groups. All rats were challenged intratracheally with alginate embedded PA and the treatments with herbal medicine started on the same day of challenge. The drugs were administered subcutaneously once a day for ten days and the control group was treated with sterile saline. The rats were sacrificed two weeks after challenge. RESULTS: Significantly improved lung bacterial clearance (P < 0.05, P < 0.01) and milder macroscopic lung pathology (P < 0.005) were found in the two treated groups compared to the control group. In the SH treated group, the neutrophil percent in the peripheral blood leukocytes (P < 0.05), the anti-PA IgG level in serum (P < 0.05), the incidence of lung abcesses (P < 0.005) and the incidence of acute lung inflammation (P < 0.05) were significantly lower than in the control group. The RAS treatment reduced fever (P < 0.05), decreased the incidence of lung abcesses (P < 0.005) and lung mast cell number (P < 0.05), and lowered anti-PA IgG1 level in serum (P < 0.05) when compared to the control group. The anti-PA bacterial activity test in SHL was weakly positive whereas in RAS it was negative. CONCLUSION: The treatment with both herbal medicines could increase the resistance of the rats against PA lung infection and they therefore might be potential promising drugs for stimulation of the immnune system in CF patients with chronic PA lung infection. PMID- 12901630 TI - Endogenous heme oxygenase/carbon monoxide system mediates lipopolysaccharide induced intussusception in rats. AB - OBJECTIVES: To investigate the role of endogenous heme oxygenase (HO)/carbon monoxide (CO) system in regulating the process of intussusception (IN) induced by administration of lipopolysaccharide (LPS) in rats. METHODS: IN model of rats were induced by lipopolysaccharide. HO activity was determined by the amount of bilirubin formation which was measured with a double-beam spectrophotometer, and HbCO formation was measured by CO-oximeter. RESULTS: The results showed that LPS (10 mg/kg) caused IN in up to 40% of the rats at 6 h after each treatment of LPS. The incidence of IN were significantly increased by 50% (P < 0.05) and by 83.2% (P < 0.01) in HO substrate (heme-L-lysinate)-treated rats and in exogenous CO treated rats, respectively; but it was sigificantly decreased by 41.8% (P < 0.05) after administration of ZnDPBG, an inhibitor of heme oxygense (HO) activity. Furthermore, LPS increased HO activity, HbCO formation cGMP content within colic smooth muscle and the plasma level of cGMP, and these parameters were significantly elevated by 62.6% (P < 0.01), 40.0% (P < 0.01), 49.3% (P < 0.05) and 38.9% (P<0.05), respectively, compared with LPS-non-IN rats. CONCLUSION: It is suggested that endogenous HO/CO system plays an important role in the process of IN induced by LPS, and inhibition of HO activity may decrease the formation of IN. PMID- 12901631 TI - Expression of intercellular adhesion molecule in lung tissues of experimental acute lung injury and the affect of Rhubarb on it. AB - OBJECTIVE: To approach the relation and the possible mechanism between the expression of intercellular adhesion molecule (ICAM-1) mRNA and acute lung injury (ALI) and the mechanisms of rhubarb in the prevention and treatment of the lung injury. METHODS: Lipopolysaccharide (LPS) was injected into the sublingual vein of male Wistar rats to perform ALI animal model. The rats were divided into 4 groups: LFS group, control group, rhubarb group and dexamethasone group. Macroscopic and histopathological examinations were performed and biological markers were measured for the lung specimens. The markers included lung wet/dry weight, the rate of neutrophils and protein content in the pulmonary alveolar lavage fluid, pulmonary vascular permeability and pulmonary alveolar permeability index. Molecular hybridization method was used to determine the expression of ICAM-1 mRNA. RESULTS: In the lung tissues, the ICAM-1 mRNA expression was increased in the endothelial cells of pulmonary veins and capillaries, rhubarb and dexamethasone had the action of decreasing the expression. The light reflex value in the gray scale scanning showed that in the comparison between the LPS and the control group, the gray scale value of the lung tissues in ALI was significantly increased, thus the light reflex value was markedly decreased (P < 0.01), demonstrating the expression of ICAM-1 mRNA was increased. In comparison with the LPS group, dexamethasone and rhubarb could decrease the gray scale value of the lung tissue significantly, thus the light reflex value was elevated (P < 0.01, P< 0.05); the corresponding pathologic change of lung tissue and the biological markers of the lung injury were significantly decreased or ameliorated. CONCLUSIONS: The increase of the expression of ICAM-1 mRNA in the lung tissues of ALI plays the roles in ALI. The application of rhubarb and dexamethasone can decease the expression and ameliorate the lung damage; its mechanism is possibly via the inhibition of ICAM-1 mRNA expression. PMID- 12901633 TI - Cloning of encoding sequence for a histone H1 from Chinese placenta genome DNA. PMID- 12901634 TI - Conservative management of twin pregnancy with single fetal death. AB - OBJECTIVE: Analysis of the cause of intrauterine death of one fetus of twin and to evaluate the outcome of conservative management of twin pregnancy with single fetal death. METHOD: A retrospective review was undertaken on PUMC hospital records of women who delivered twin between Jan. 1987 and Dec. 1998. RESULT: During the recent 12 years, 99 pregnant women were documented as twin pregnancies. Of the twin pregnancies during this period, 13 were complicated by a single intrauterine death. Four women suffered this complication during their first trimester. In the remain 9 cases one fetus died during second or third trimester. The cesarean section was given in three pregnant women soon after a single intrauterine death because of the survival fetal distress. No consumptive coagulopathy was apparent in all 13 pregnant women. It is wise that the pregnancy was managed conservatively, with regular coagulation parameters obtained. CONCLUSION: Our results support conservative management in twin pregnancies complicated by single fetal death. PMID- 12901632 TI - Role of endogenous carbon monoxide in endotoxin shock. AB - OBJECTIVE: To study the role of endogenous carbon monoxide (CO) in endotoxin shock. METHODS: The changes of CO levels and the effects of zinc protoporphyrin IX (ZnPP), an inhibitor of heme-oxygenase (HO), in endotoxin shock and the efficacy of hemin, an inducer of HO were investigated. RESULTS: The plasma CO levels were found to be significantly increased during the course of endotoxin shock. Injection of ZnPP was shown to abrogate the endotoxin-induced hypotension and metabolic derangements markedly. Administration of hemin to healthy rabbits revealed the hypotension and metabolic derangements similar to the animals given endotoxin. CONCLUSION: CO is a newly found endogenously produced mediator which may play an important role in the pathogenesis of endotoxin shock. PMID- 12901635 TI - Comparison the efficiency of different surgical procedures for urinary stress incontinence. AB - OBJECTIVE: To analyse the efficiency of two operations for treatment of urinary stress incontinence (USI). METHODS: The clinical data of 95 USI cases or accompany USI treated by surgical procedures (Kelly operation 63 cases, Colposuspension 32 cases) was analysed retrospectively in our department. RESULTS: The cure rates during three months after operation were 92.9% in Colposuspension group and 68.9% in Kelly operation group (P < 0.05), and decreased to 73.7% and 57.5% one year after operation in Colposuspension group and Kelly operation group respectively (P > 0.05). Abnormal senses rates of sexual intercourse was 3.7% in Colposuspension and 7.3% in Kelly operation group. The length of retaining Foley catheter after Colposuspension was more than Kelly operation. CONCLUSION: The cure rate during three months after Colposuspension is better than Kelly operation. But the cure rates during one year after surgery are the same in two groups. PMID- 12901636 TI - Surgical treatment of spondylolisthesis with SOCON instrumentation. AB - OBJECTIVE: To observe the clinical outcomes of using a new instrumentation SOCON system in the treatment of degenerative spondylolisthesis. METHODS: Retrospective clinical and roentgenograph review of 21 patients who suffered from degenerative spondylolisthesis with spinal stenosis treated by decompression, posterolateral intertransverse arthrodesis, and with transpedicle instrumentation of SOCON system. RESULTS: Nineteen of 21 patients were completely recovery from their preoperative symptoms, 17 of 18 cases with grade one slippage reduced to normal spine alignment, 2 cases of grade two slippage fully reduced, and one case of grade two spondylolisthesis got 70% reduction. Post-operative satisfactory rate was 90.5%. Pain relief was 90.5%. Neither infection nor neurologic complication occurred in this series. CONCLUSION: Our short time follow-up and limited cases showed satisfactory preliminary result of surgical treatment of spondylolisthesis with SOCON instrumentation. PMID- 12901637 TI - Overview for the diagnosis and treatment of gallbladder carcinoma. AB - OBJECTIVE: To improve the recognition of diagnosis and treatment of gallbladder cancer. METHODS: Retrospective analysis of 52 cases of gallbladder carcinoma in our hospital from 1988 to 1998. RESULTS: Preoperative diagnostic rate was 90.3%, of which 12 cases (23%) were early stage of carcinoma. The total operation resection rate was 55.8%, for which only 17.8% were advance stage of carcinoma. CONCLUSION: The early diagnosis is the key factor of increasing treatment successful rate. The following are the symptoms that raise our special attention to carcinoma of galbladder: (1) Age over 50 have recurrent cholecystitis and with past history of gallstone; (2) Congenital malformation of bile duct; (3) Local thickening and irregularity of gallbladder wall; (4) Polypoid lesion larger than 1 cm inside gallbladder; (5) Atrophic gallbladder; (6) Intraluminal stone of gallbladder does not move when change in body position; (7) Regional lymph node enlargement. PMID- 12901638 TI - Study of the relationship between psoriasis and the polymorphic site C677T of methylenetetrahydrofolate reductase. AB - OBJECTIVE: In order to investigate 5,10-methylenetetrahydrofolate reductase (MTHFR)'s polymorphic changes in psoriasis vulgaris. METHODS: We detected mutation of site C677V of MTHFR in 39 psoriastics by PCR-RFLP. RESULTS: Genotype frequencies of the psoriastics were C/C=20.15%, C/T=48.72% and T/T=30.77%; the allelic frequencies were C=0.4487 and T=0.5513. Homozygous mutant (TT) of the psoriastics was significantly different from the normal control group by X2 test. CONCLUSION: C677V mutant of MTHFR might be related with psoriasis. PMID- 12901639 TI - The first case of Protothecosis zopfii in China. AB - OBJECTIVE: Report of first case of Protothecosis zopfii in China and causes the skin infection in the world. METHOD: By clinical and laboratory examinations to confirm the diagnosis and the response to treatment. By the review of literatures to confirm the first case of human skin infection in the world. RESULT: From the literatures and the clinical picture, it is confirmed that this is the first case report of Protothecosis zopfii of skin in the world. CONCLUSION: The first case of Protothecosis zopfii in human being was reported and successfully treated with local infiltration of Diflucan (fluconazole) 3 ml (2 mg/ml)/week x 4. PMID- 12901640 TI - The initial immunological research of laryngeal cancer. PMID- 12901642 TI - A retrospective study of 60 hospitalized cases with pemphigoid. PMID- 12901641 TI - Preparation of anti-idiotypic antibodies specific for anti-HEL and analysis of their functional mimicry. AB - OBJECTIVE: This study is to investigate the functional mimicry by using anti idiotypic antibodies of enzymes. METHODS: Monoclonal anti-idiotypic antibodies against anti-HEL (hen egg-white lysozyme, HEL) antibodies were obtained by fusion of Sp2/0 myeloma cells with spleen cells of syngeneic mice immunized with monoclonal anti-HEL antibodies against HEL's different antigenic epitopes. Then bacteriolysis of the anti-idiotypic antibodies were observed. RESULTS: Eight hybridomas strains secreting anti-idiotypic antibodies were selected and characterized. It was shown that two of eight anti-idiotypic antibodies secreted by two hybridomas (1A10C9 and 2A11C1B3) could mimic HEL catalytic activity to lyse Micrococcus lysodeikticus and that the catalytic effect of mixed anti idiotypic antibodies of 1A10C9 and 2A11C1B3 was stronger than that of one of them, but less than HEL. CONCLUSION: The results demonstrated that the anti idiotypic antibodies that could mimic enzyme activity existed in the idiotype network during anti-enzymatic immune response. PMID- 12901643 TI - Relationship between CD44v6 expression and metastatic potential and prognosis in gastric carcinoma. PMID- 12901644 TI - HLA-A*02 subtype distribution of Han nationality in northern China. PMID- 12901645 TI - Effects of propofol compared to thiopentoneon preventing hemodynamic response to tracheal intubation. PMID- 12901646 TI - Relationship between plasma motilin and effect of oral erythromycin on gallbladder contraction in patients of functional dyspepsia. PMID- 12901647 TI - Prognostic significance of heart rate variability in post-myocardial infarction patients. PMID- 12901648 TI - Natural background concentrations of nutrients in streams and rivers of the conterminous United States. AB - Determining natural background concentrations of nutrients in watersheds in the developed world has been hampered by a lack of pristine sampling sites covering a range of climatic conditions and basin sizes. Using data from 63 minimally impacted U.S. Geological Survey reference basins, we developed empirical models of the background yield of total nitrogen (TN) and total phosphorus (TP) from small watersheds as functions of annual runoff, basin size, atmospheric nitrogen deposition rate, and region-specific factors. We applied previously estimated in stream loss rates to yields from the small watershed models to obtain estimates of background TN and TP yield and concentration throughout the stream/river network in 14 ecoregions of the conterminous United States. Background TN concentration varies from less than 0.02 mg L(-1) in the xeric west to more than 0.5 mg L(-1) along the southeastern coastal plain. Background TP concentration varies from less than 0.006 mg L-(1) in the xeric west to more than 0.08 mg L(-1) in the great plains. TN concentrations in U.S. streams and rivers currently exceed natural background levels by a much larger factor (6.4) than do TP concentrations (2.0). Because of local variation in runoff and other factors, the range of background nutrient concentrations is very large within some nutrient ecoregions. It is likely that background concentrations in some streams in these regions exceed proposed nutrient criteria. PMID- 12901649 TI - A water resources threshold and its implications for food security. AB - Cereal import has played a crucial role in compensating local water deficit. A quantitative account of water deficit and cereal import relations therefore is of significance for predicting future food import demand and formulating corresponding national and international policies. On the basis of data for countries in Asia and Africa, we estimated a water resources threshold with respect to cereal import. Below the threshold, the demand for cereal import increases exponentially with decreasing water resources. There appeared to be a declining trend in the threshold, from 2000 m3/(capita year) in the early 1980s to 1500 m3/(capita year) by the end of the 1990s. Until recently, most countries below the threshold were oil-rich and thus were able to afford cereal import. However, the next 30 yr may see many poor and populous countries dropping below the threshold in association with their rapid population growth and the depletion of fossil groundwater. Water deficit-induced food insecurity and starvation could intensify because cereal import may not be affordable for these countries. PMID- 12901650 TI - Organic and elemental carbon measurements during ACE-Asia suggest a longer atmospheric lifetime for elemental carbon. AB - During the ACE-Asia intensive field campaign (March 14-April 20, 2001), PM1.0 organic (OC) and elemental carbon (EC) concentrations were measured onboard the NOAA R/V Ronald H. Brown over the Northwest Pacific Ocean using a semi-continuous automated carbon analyzer downstream of a carbon-impregnated filter denuder. This OC and EC measurement achieved a mean time resolution of about 200 min over the Pacific Ocean, substantially lower than that achieved previously (24 h). The semi continuous measurements, in which the adsorption artifact was substantially reduced using the denuder, showed good agreement with integrated artifact corrected measurements made without a denuder. Mean particulate OC and EC concentrations were 0.21 and 0.09, 0.70 and 0.29, 1.00 and 0.27, and 2.43 and 0.66 microg of C m(-3) over the background Pacific Ocean, Asian-influenced Pacific Ocean, offshore of Japan, and Sea of Japan, respectively. On April 11, 90 min average OC and EC concentrations peaked at 4.0 and 1.3 microg of C m(-3), respectively, offshore of Korea over the Sea of Japan. The OC/EC ratio of 3.7 over the Sea of Japan and offshore of Japan was substantially higher than that of 2.5 over the Asian-influenced Pacific Ocean, even though backward air mass trajectories put the "Asian-influenced Pacific Ocean" sample downwind. The OC/EC ratio decreased with increasing time since the air mass encountered the source regions of China, Japan, and Korea. This suggests a longer atmospheric residence time for EC than for OC. PMID- 12901651 TI - Effect-directed identification of oxygen and sulfur heterocycles as major polycyclic aromatic cytochrome P4501A-inducers in a contaminated sediment. AB - Heterocyclic polyaromatic compounds, including dinaphthofurans, 2-(2 naphthalenyl)benzothiophene, methylated chrysene, and benz[a]anthracene, were identified and confirmed as major cytochrome P4501A (CYP1A)-inducing compounds in a contaminated sediment close to the industrial site of Bitterfeld (Germany). Identification was achieved by the application of an effect-directed fractionation and analysis approach. This approach comprised the combination of a rainbow trout liver cell line (RTL-W1) bioassay to select for CYP1A-inducing effects by measuring 7-ethoxyresorufin-O-deethylase activity, a multistep fractionation procedure, and various methods of chemical characterization. The identified nonpriority pollutants were found to be significantly more potent than the reference compound, benzo[a]pyrene, and among the most potent polycyclic inducers known. On the basis of the history of industrial activity at the contaminated site, the heterocyclic compounds identified in this study are thought to stem from 2-naphthol production. 2-Naphthol is one of the most high tonnage products of the aniline dye industry in general, thereby indicating the potential environmental relevance of the identified heterocyclic aromatic compounds. To date, however, no or very little knowledge exists about their occurrence, fate, and biological effects. PMID- 12901652 TI - Dioxins, furans, and non-ortho PCBs in Canadian Arctic seabirds. AB - This is the first account of PCDDs, PCDFs, and non-ortho PCBs in Canadian Arctic seabirds. Livers and eggs of thick-billed murres, northern fulmars, and black legged kittiwakes were collected in 1975 and 1993 from Prince Leopold Island in Lancaster Sound, Canada. Detectable concentrations of PCDDs, PCDFs, and non-ortho PCBs were found in all the Arctic seabird samples analyzed. Of the PCDD congeners assayed, only 2,3,7,8-substituted PCDDs were detected in the samples, whereas non 2,3,7,8-substituted PCDFs were found in addition to 2,3,7,8-substituted PCDFs in some of the samples. The predominant PCDD/F congener found in the livers of all three species was 2,3,4,7,8-PnCDF, both in 1975 and 1993. Concentrations of most dioxins and furans decreased in the fulmars and kittiwakes between 1975 and 1993 but increased in the murres. Of the non-ortho PCBs measured, PCB-126 occurred in the highest concentrations and contributed the majority of the non-ortho PCB-TEQ in all three species in both years. The highest concentrations of dioxins and furans as well as the highest TEQ values were found in the northern fulmar livers in both 1975 and 1993. Concentrations of some of the PCDDs and PCDFs are among the highest reported for Canadian Arctic biota. PMID- 12901653 TI - Geographical, temporal, and species variation of the polyether toxins, azaspiracids, in shellfish. AB - Azaspiracid Poisoning (AZP) is a new toxic syndrome that has caused human intoxications throughout Europe following the consumption of mussels (Mytilus edulis), harvested in Ireland. Shellfish intoxication is a consequence of toxin bearing microalgae in the shellfish food chain, and these studies demonstrated a wide geographic distribution of toxic mussels along the entire western coastal region of Ireland. The first identification of azaspiracids in other bivalve mollusks including oysters (Crassostrea gigas), scallops (Pecten maximus), clams (Tapes phillipinarium), and cockles (Cardium edule) is reported. Importantly, oysters were the only shellfish that accumulated azaspiracids at levels that were comparable with mussels. The highest levels of total azaspiracids (microg/g) recorded to-date were mussels (4.2), oysters (2.45), scallops (0.40), cockles (0.20), and clams (0.61). An examination of the temporal variation of azaspiracid contamination of mussels in a major shellfish production area revealed that, although maximum toxin levels were recorded during the late summer period, significant intoxications were observed at periods when marine dinoflagellate populations were low. Although human intoxications have so far only been associated with mussel consumption, the discovery of significant azaspiracid accumulation in other bivalve mollusks could pose a threat to human health. PMID- 12901654 TI - Calcium losses from a forested catchment in south-central Ontario, Canada. AB - Mass balance studies at a number of calibrated watersheds in eastern North America suggest that large losses of Ca from soil have occurred due to acid deposition. However, there is considerable controversy over whether losses have occurred from the exchangeable pool or whether there are other sources of Ca that have not been considered. Mass balance calculations at a small, calibrated catchment (PC1) in south-central Ontario also indicate that large losses of Ca have occurred over the past 2 decades. If the exchangeable Ca pool has declined by approximately 40% between 1983/1984 and 1998/1999 but the exchangeable Mg pool has remained relatively stable, these changes should be evident in streamwater chemistry. The slope of the buffer curve relating Ca to (sigma)acid anions (F(Ca)) decreased significantly over the study period. We estimate that F(Ca) decreased by 12-24% between 1983 and 1999, although the level of decrease is affected by changes in hydrology. During the same period, there was no significant change in F(Mg), and the annual volume-weighted Ca:Mg (equiv/equiv) ratio in streamwater decreased from approximately 2.8 to approximately 2.1. Measured changes in streamwater chemistry translate into Ca losses from the exchangeable pool of approximately 250-570 mequiv/m2 over the study period, which is similar to estimated losses based on soil measurements (approximately 425 mequiv/m2) and mass balance calculations (approximately 380-430 mequiv/m2). The magnitude of Ca loss cannot be explained by decreases in Ca deposition or decreased Ca weathering. Large Ca losses from the soil exchangeable pool have serious implications for future forest health and the recovery of streamwater from acid deposition. PMID- 12901655 TI - Identifying the effect of polar constituents in coal-derived NAPLs on interfacial tension. AB - Interfacial tension, which is a critical variable affecting multiphase flow of nonaqueous phase liquids (NAPLs) in the subsurface, varies greatly with the composition of complex NAPLs recovered from field sites. Much of this variability stems from the presence of acid and base molecules in the NAPL mixture. The interfacial tension and acid and base concentration in six coal-derived NAPLs were measured. Creosotes generally have lower interfacial tensions due to their higher concentrations of organic acid and base macromolecules as compared with coal tar samples. Interfacial tension is a function of pH, with lower values measured at pH greater than approximately 9. At a neutral pH, the interfacial tensions are inversely proportional to the total acid concentration. Asphaltenes in these coal-derived NAPLs account for most of the acid and base constituents. It is found in this study that acid and base numbers are valuable indicators of interfacial tension and, therefore, the capillary phenomena associated with multiphase flow behavior of NAPLs in the subsurface. PMID- 12901656 TI - Shifts in biodegradation kinetics of the herbicides MCPP and 2,4-D at low concentrations in aerobic aquifer materials. AB - Biodegradation kinetics of two phenoxy acid herbicides, MCPP [(+/-)-2-(4-chloro-2 methylphenoxy)propanoic acid; mecoprop] and 2,4-D [2,4-dichlorophenoxyacetic acid] were studied in laboratory batch microcosms at low concentrations (0.025 100 microg/L) using 14C technique with sediments and groundwater from a shallow aerobic sandy aquifer. Below a certain threshold concentration of approximately 1 microg/L for 2,4-D and 10 microg/L for MCPP, the biodegradation followed first order nongrowth kinetics, and no adaptation was observed within the experimental period of 341 d. Half-lifes for ultimate degradation were 500 d for 2,4-D and 1100 d for MCPP at 10 degrees C in unpolluted aquifer sediment in this environmentally relevant concentration regime. Above the threshold concentrations, the biodegradation rate accelerated gradually due to selective growth of specific biomass, which was ascertained from 14C most probable number enumerations of specific phenoxy acid degraders. Atthe highest concentration tested (100 microg/ L), specific degraders increased from 10(-1) to 10(5) cells/g during the experiment, and half-lifes after adaptation decreased to approximately 5 d. The enhanced rate of degradation by adapted systems was maintained during degradation of the last residuals measured to less than 0.1 microg/L. In situ long-term preexposure of the aquifer sediment also resulted in significant higher degradation rates of the phenoxy acids. PMID- 12901657 TI - Bromate in chlorinated drinking waters: occurrence and implications for future regulation. AB - Bromate is a contaminant of commercially produced solutions of sodium hypochlorite used for disinfection of drinking water. However, no methodical approach has been carried out in U.S. drinking waters to determine the impact of such contamination on drinking water quality. This study utilized a recently developed method for quantitation of bromate down to 0.05 microg/L to determine the concentration of bromate present in finished waters that had been chlorinated using hypochlorite. Forty treatment plants throughout the United States using hypochlorite in the disinfection step were selected and the levels of bromate in the water both prior to and following the addition of hypochlorite were measured. The levels of bromate in the hypochlorite feedstock were also measured and together with the dosage information provided by the plants and the amount of free chlorine in the feedstock, it was possible to calculate the theoretical level of bromate that would be imparted to the water. A mass balance was performed to compare the level of bromate in finished drinking water samples to that found in the corresponding hypochlorite solution used for treatment. Additional confirmation of the source of elevated bromate levels was provided by monitoring for an increase in the level of chlorate, a co-contaminant of hypochlorite, atthe same point in the treatment plant where bromate was elevated. This study showed that bromate in hypochlorite-treated finished waters varies across the United States based on the source of the chemical feedstock, which can add as much as 3 microg/L bromate into drinking water. Although this is within the current negotiated industry standard that allows up to 50% of the maximum contaminant level (MCL) for bromate in drinking water to be contributed by hypochlorite, it would be a challenge to meet a tighter standard. Given that distribution costs encourage utilities to purchase chemical feedstocks from local suppliers, utilities in certain regions of the United States may be put at a distinct disadvantage if future lower regulations on bromate levels in finished drinking water are put into place. Moreover, with these contaminant levels it would be almost impossible to lower the maximum permissible contribution to bromate in finished water from hypochlorite to 10% of the MCL, which is the norm for other treatment chemicals. Until this issue is resolved, it will be difficult to justify a lowering of the bromate MCL from its current level of 10 to 5 microg/L or lower. PMID- 12901658 TI - Removal of two polycyclic musks in sewage treatment plants: freely dissolved and total concentrations. AB - In the current study, the removal of slowly degradable hydrophobic chemicals in sewage treatment plants (STPs) has been evaluated with emphasis on the combination of free and total concentration data. Free and total concentrations of two polycyclic musks were determined in each compartment of four STPs. The free concentration of the polycyclic musks remains virtually constant throughout all the compartments of the STPs with values between 0.21 and 0.57 microg/L for AHTN and between 0.79 and 2.0 microg/L for HHCB. Total concentrations of these fragrances are highly dependent on the volatile solids in a given compartment resulting in much more variation with values between 0.42 and 92 microg/L for AHTN and between 1.25 and 258 microg/L for HHCB. It is concluded that free concentrations of these hydrophobic chemicals in the compartments of STPs are mostly biodegradation mediated, while total concentrations are mediated by the concentration of solids. The combination of measurements of free and total concentrations can improve estimations regarding removal efficiency and removal pathways. PMID- 12901659 TI - Interactions among contaminants and nutritional lipids during mobilization by digestive fluids of marine invertebrates. AB - Coastal sediments contain complex mixtures of hydrophobic compounds including organic contaminants such as polycyclic aromatic hydrocarbons and biogenic compounds such as cholesterol and phospholipids. Within the guts of benthic invertebrates, these mixtures are subjected to digestive, chemical conditions that can be rich in surfactants and proteinaceous material. Using in vitro incubations as proxy for digestive exposure, we studied the solubilization of binary mixtures of nutritional and contaminant lipids into artificial seawater and six marine invertebrate gut fluids (Molpadia intermedia, Cucumaria frondosa, Arenicola marina, Arenicola brasiliensis, Parastichopus californicus, and Nereis virens). For animals with surfactant micelles or high protein concentrations, solubilization interactions were frequent. For example, in Arenicola marina gut fluid, benzo(a)pyrene enhanced the solubilization of hexadecane (491% of the compound alone) and palmitic acid (130%) but hindered cholesterol (83%). Benzo(a)pyrene concentrations increased in gut fluids in the presence of cholesterol (137% of BaP alone), phenanthrene (154%), lecithin (140%), and hexadecanol (232%). In A. marina gut fluid, dilution with seawater indicated that these enhancements occur only when micelles are present. Sediment-water partitioning models, used to predict the bioavailability of hydrophobic organic chemicals, do not account for such interactions between solubilizates (compounds solubilized in micelles). However, for animals exposed via a digestive tract containing micelles or high protein concentrations, digestive bioavailability and perhaps bioaccumulation are likely influenced by these interactions. PMID- 12901660 TI - Chemical-specific representation of air--soil exchange and soil penetration in regional multimedia models. AB - In multimedia mass-balance models, the soil compartment is an important sink as well as a conduit for transfers to vegetation and shallow groundwater. Here a novel approach for constructing soil transport algorithms for multimedia fate models is developed and evaluated. The resulting algorithms account for diffusion in gas and liquid components; advection in gas, liquid, or solid phases; and multiple transformation processes. They also provide an explicit quantification of the characteristic soil penetration depth. We construct a compartment model using three and four soil layers to replicate with high reliability the flux and mass distribution obtained from the exact analytical solution describing the transient dispersion, advection, and transformation of chemicals in soil layers with different properties but a fixed boundary condition at the air-soil surface. The soil compartment algorithms can be dynamically linked to other compartments (air, vegetation, groundwater, surface water) in multimedia fate models. We demonstrate and evaluate the performance of the algorithms in a model with applications to benzene, benzo[a]pyrene, MTBE, TCDD, and tritium. PMID- 12901661 TI - Dioxins in aquatic sediment and soil in the Kanto region of Japan: major sources and their contributions. AB - Major sources of dioxins (tetra- to octa-chlorinated dibenzo-p-dioxins and dibenzofurans) were quantitatively identified, and the contributions of each source were estimated in aquatic surface sediment and soil in the Kanto region of Japan, where a major part of the municipal, industrial, and agricultural activities in Japan take place. Detailed chromatographic peak-specific data corresponding to all dioxin compounds were used throughout the analysis to achieve source specificity. Four major sources were identified by using principal component analysis. Three of them were combustion processes, impurities in pentachlorophenol, and impurities in the diphenyl ether herbicide CNP. One, characterized by polychlorinated dibenzo-p-dioxins with 1,2,6,9-chlorine substitution, remained unattributed to any known source or transformation process. The composition of the unattributed source resembled those reported in sediment samples free from anthropogenic impacts. Contributions from the four major sources were estimated by using a mass balance receptor model, and the source contributions to each homologue varied among samples. Tokyo Bay sediment was characterized by the highest relative contribution from pentachlorophenol impurities in the region. The unattributed source contribution was most prominent in the Lake Kasumigaura sediment. The results indicated considerable contributions in aquatic surface sediment in the studied region from dioxin impurities in herbicides used in the past. PMID- 12901662 TI - In-situ characterization of soil-water content using gas-phase partitioning tracer tests: field-scale evaluation. AB - Field-scale tests were performed to evaluate the effectiveness of the gas-phase partitioning tracer method for in-situ measurement of soil-water content. The tracer tests were conducted before and after a controlled infiltration event to evaluate performance at two water contents. Nonpartitioning (sulfur hexafluoride) and water-partitioning (difluoromethane) tracers were injected into the test zone, and their effluent breakthrough curves were analyzed using the method of moments to calculate retardation factors for difluoromethane. Soil-water contents estimated using the tracer data were compared to soil-water contents obtained independently using gravimetric core analysis, neutron scattering, and bore-hole ground penetrating radar. For the test conducted under drier soil conditions, the soil-water content estimated from the tracer test was identical to the independently measured values of 8.6% (equivalent to water saturation of 23%). For the test conducted under wetter soil conditions, the tracer test derived soil water content was 81% of the independently measured values of 12.2% (equivalent to water saturation of 32%). The reduced efficacy at the higher soil-water content may reflectthe impact of advective and/ or diffusive mass transfer constraints on gas-phase transport. The results presented herein indicate that the partitioning tracer method is an effective technique to measure soil-water content at the field scale, especially for sites with moderate to low soil-water contents. PMID- 12901663 TI - Sampling atmospheric carbonaceous aerosols using an integrated organic gas and particle sampler. AB - Measurement of particle-bound organic carbon (OC) may be complicated by sampling artifacts such as adsorption of gas-phase species onto particles or filters or evaporation of semivolatile compounds off the particles. A denuder-based integrated organic gas and particle sampler (IOGAPS), specifically designed to minimize sampling artifacts, has been developed to sample atmospheric carbonaceous aerosols. IOGAPS is designed to first remove gas-phase chemicals via sorption to the XAD-coated denuder, and subsequently particles are trapped on a quartz filter. A backup sorbent system consisting of sorbent- (XAD-4 resin) impregnated filters (SIFs) was used to capture the semivolatile OC that evaporates from the particles accumulated on the upstream quartz filter. A traditional filter pack (FP) air sampler, which uses a single quartz filter to collect the particles, was employed for comparison in this study. Elemental and organic carbon were determined from filter punches by a thermal optical transmittance aerosol carbon analyzer. Field measurements show that there was no significant difference between the elemental carbon concentrations determined by the FP and IOGAPS, indicating that particle loss during the transit through the denuder tube was negligible. Compared with the OC determined by FP (3.9-12.6 microg of C/m3), the lower OC observed on the quartz filter in the IOGAPS (2.2 6.0 microg of C/m3) was expected because of the removal of gas-phase organics by the denuder. Higher semivolatile organic carbon (SVOC) on the backup SIFs during the night (1.24-8.43 microg of C/m3) suggests that more SVOC, emitted from primary sources or formed as secondary organic compounds, partitions onto the particles during the night because of the decreased ambient temperature. These data illustrate the utility of an IOGAPS system to more accurately determine the particle-bound OC in comparison to FP-based systems. PMID- 12901664 TI - Incineration of different types of medical wastes: emission factors for particulate matter and heavy metals. AB - Previously published results for emission factors of medical waste incineration do not include enough information about the incinerated waste composition. This paper reports the first emission factors estimated for particulate matter, As, Cd, Cr, Pb, Mn, Hg, and Ni, considering that medical waste is segregated in different types according to Portuguese legislation. The main purpose was to evaluate the influence of incinerated waste composition and segregation practice on emission factors. One "controlled-air" incinerator without air pollution control devices was used for the incineration either of mixtures with a defined composition or of a specific waste type. Previously published emission factors are not associated with the composition of the incinerated mixture, and the results showed that the usefulness of those emission factors is very doubtful. The existence of different waste classifications also reduces the usefulness of previously published results. To protect human health, appropriate equipment to control atmospheric pollutants must be used, since the legal limits for pollutant concentrations were strongly surpassed (226 times higher than the limit for Hg), with risks for patients and workers of the hospital and exposed population. It was concluded that rigorous segregation practices and adequate management methodologies allow reducing 80% of the amount of wastes that must be incinerated, practically eliminating Hg and Pb emissions and reducing those of PM, As, Cd, Cr, Mn, and Ni, respectively, 98, 90, 92, 84, 77, and 92%. PMID- 12901665 TI - Adsorption of the endocrine-active compound estrone on microfiltration hollow fiber membranes. AB - Results of studies reported here show that adsorption could result in considerable accumulation of hormones on hydrophobic hollow fiber membrane surfaces during filtration of trace-hormone-containing feed solutions with a linear adsorption isotherm applicable over the majority of the estrone concentration range examined (2.6-154 ng/L). Models based on both diffusion and surface reaction limitation were used to describe the kinetics of estrone adsorption to the membranes tested. Results indicate that the rate of adsorption of estrone to the hollow fiber membranes was limited principally by surface reaction rate rather than the rate of diffusive transport to membrane surface sites. Both adsorption and desorption kinetics were satisfactorily described by pseudo-first-order expressions. These results are of environmental significance, especially in drinking water applications, where contaminants such as natural and synthetic hormones may accumulate on the membranes and desorb during backwashing and membrane cleaning. PMID- 12901666 TI - Application of electron beam for the reduction of PCDD/F emission from municipal solid waste incinerators. AB - The electron-beam technology was applied to reduce the emission of polychlorinated dibenzo-p-dioxins (PCDD) and polychlorinated dibenzofurans (PCDF) in a flue gas of 1000 m(3)N/h from the municipal solid waste incinerator (MSWI) at a temperature of 200 degrees C. More than 90% decomposition of PCDD/Fs was obtained using an electron accelerator at a dose of 14 kGy. The decomposition was initiated through reactions with OH radicals produced by the irradiation of flue gases, followed by oxidation such as the ring cleavage of the aromatic ring, the dissociation of ether bond, and dechlorination. The cost analysis estimated that the electron-beam system can cut the annualized cost by approximately 50% for the treatment of PCDD/Fs in a pre-dusted MSWI flue gas as compared with a bag-filter system when operating on electricity generated from an incineration. Electron beam technology is an economically and technologically useful method for reducing PCDD/Fs in an incineration flue gas. PMID- 12901667 TI - Comparison performances of membrane bioreactor and conventional activated sludge prcesses on sludge reduction induced by Oligochaete. AB - Pilot-scale experiments were carried out to compare sludge reduction induced by Oligochaete in a submerged membrane bioreactor (MBR) and a conventional activated sludge (CAS) reactor for 345 d. Worm growth in the CAS reactor was much better than in the MBR. The average worm density of the aeration tank in the CAS reactor was 71 total worms/mg of volatile suspended solids (VSS), much higher than that in the MBR (10 total worms/mg of VSS). Worms did not naturally produce in the MBR, and the dominant worm type in the MBR depended on sludge inoculation from the CAS reactor. Only two types of worms were found in the MBR, Aeolosoma hemprichicii and Nais elinguis. Worm presence and disappearance in the MBR alternated. Worms in the CAS reactor occurred nearly throughout the operating period and were continuously maintained at over 30 total worms/mg of VSS in the aeration tank for 172 d. Three types of worm were found in the CAS reactor, A. hemprichicii, Pristina aequiseta, and N. elinguis, but P. aequiseta was present only occasionally. The alternating dominance of worm types in both reactors changed between Aeolosoma and Nais, and the time of Aeolosoma dominance was longer than that of Nais dominance. Worm growth in the MBR contributed to neither sludge reduction nor improvement of sludge settling characteristics because of low density. But worm presence and bloom in the CAS reactor greatly decreased sludge yield and improved sludge settling characteristics at high density. Both the average sludge yield (0.17 kg of suspended solids (SS)/kg of chemical oxygen demand removed (CODremoved)) and sludge volume index (60 mL/g) in the CAS reactor were much lower than those in the MBR (0.40 kg of SS/kg of CODremoved and 133 mL/g). Nais had more potential for sludge reduction than Aeolosoma. Worm growth had little impact on effluent quality in the MBR but affected effluent quality very much in the CAS reactor. PMID- 12901668 TI - Acetylene inhibition of trichloroethene and vinyl chloride reductive dechlorination. AB - Kinetic studies reported here have shown that acetylene is a potent reversible inhibitor of reductive dehalogenation of trichloroethene (TCE) and vinyl chloride (VC) by a mixed dehalogenating anaerobic culture. The mixed culture was enriched from a contaminated site in Corvallis, OR, and exhibited methanogenic, acetogenic, and reductive dehalogenation activities. The H2-fed culture transformed TCE to ethene via cis-dichloroethene (c-DCE) and VC as intermediates. Batch kinetic studies showed acetylene reversibly inhibited reduction of both TCE and VC, and the levels of inhibition were strongly dependent on acetylene concentrations in both cases. Acetylene concentrations of 192 and 12 microM, respectively, were required to achieve 90% inhibition in rates of TCE and VC transformation at an aqueous concentration of 400 microM. Acetylene also inhibited methane production (90% inhibition at 48 microM) but did not inhibit H2 dependent acetate production. Mass balances conducted during the studies of VC inhibition showed that acetogenesis, VC transformation to ethene, and methane production were responsible for 52%, 47%, and 1% of the H2 consumption, respectively. The results indicate that halorespiration is the dominant process responsible for VC and TCE transformation and that dehalorespiring organisms are the target of acetylene inhibition. Acetylene has potential use as a reversible inhibitor to probe the biological activities of reductive dechlorination and methanogenesis. It can be added to inhibit reactions and then removed to permit reactions to proceed. Thus, it can be a powerful tool for investigating intrinsic and enhanced anaerobic remediation of chloroethenes at contaminated sites. The results also suggest that acetylene produced abiotically by reactions of chlorinated ethenes with zero-valent iron could inhibit the biological transformation of VC to ethene. PMID- 12901669 TI - Rate and extent of aqueous perchlorate removal by iron surfaces. AB - The rate and extent of perchlorate reduction on several types of iron metal was studied in batch and column reactors. Mass balances performed on the batch experiments indicate that perchlorate is initially sorbed to the iron surface, followed by a reduction to chloride. Perchlorate removal was proportional to the iron dosage in the batch reactors, with up to 66% removal in 336 h in the highest dosage system (1.25 g mL(-1)). Surface-normalized reaction rates among three commercial sources of iron filings were similar for acid-washed samples. The most significant perchlorate removal occurred in solutions with slightly acidic or near-neutral initial pH values. Surface mediation of the reaction is supported by the absence of reduction in batch experiments with soluble Fe2+ and also by the similarity in specific reaction rate constants (kSA) determined for three different iron types. Elevated soluble chloride concentrations significantly inhibited perchlorate reduction, and lower removal rates were observed for iron samples with higher amounts of background chloride contamination. Perchlorate reduction was not observed on electrolytic sources of iron or on a mixed-phase oxide (Fe3O4), suggesting that the reactive iron phase is neither pure zerovalent iron nor the mixed oxide alone. A mixed valence iron hydr(oxide) coating or a sorbed Fe2+ surface complex represent the most likely sites for the reaction. The observed reaction rates are too slow for immediate use in remediation system design, but the findings may provide a basis for future development of cost effective abiotic perchlorate removal techniques. PMID- 12901670 TI - Tannery wastewater treatment by sequencing batch biofilm reactor. AB - The paper reports the results of an investigation aimed to evaluate the performances of an innovative process for treating tannery wastewater. In such a process biological degradation, carried out in a sequencing batch biofilm reactor (SBBR), is combined with chemical oxidation by ozone. The treatment was carried out at laboratory scale on a real primary effluent coming from a centralized plant treating the wastewater of a large tannery district in Northern Italy. SBBR performances without and with ozonation were compared with very satisfactory results particularly in the latter instance when the recorded COD, TKN, and TSS average removals, (96%), (92%), and (98%), respectively, permitted to achieve the fixed limits enforced by Italian regulation without needing any additional polishing step. With or without ozonation, the process that resulted was characterized by a specific sludge production (0.1 kgVSS/kg CODremoved) significantly lower than the values featuring conventional biological systems (i.e., 0.3-0.5 VSS/kg CODremoved). Moreover, as in the reactor the biomass density results were very high, i.e., 98 gVSS/Lsludge, it was possible to achieve and maintain biomass concentration as high as 20 gVSS/L. PMID- 12901671 TI - Chromium remediation or release? Effect of iron(II) sulfate addition on chromium(VI) leaching from columns of chromite ore processing residue. AB - Chromite ore processing residue (COPR), derived from the so-called high lime processing of chromite ore, contains high levels of Cr(III) and Cr(VI) and has a pH between 11 and 12. Ferrous sulfate, which is used for remediation of Cr(VI) contamination in wastewater and soils via reduction to Cr(III) and subsequent precipitation of iron(III)/chromium(III) hydroxide, has also been proposed for remediation of Cr(VI) in COPR. Instead, however, addition of FeSO4 to the infiltrating solution in column experiments with COPR greatly increased leaching of Cr(VI). Leached Cr(VI) increased from 3.8 to 12.3 mmol kg(-1) COPR in 25 pore volumes with 20 mM FeSO4, reaching solution concentrations as high as 1.6 mM. Fe(II) was ineffective in reducing Cr(VI) to Cr(III) because it precipitated when it entered the column due to the high pH of COPR, while Cr(VI) in solution was transported away with the infiltrating solution. The large increase in leaching of Cr(VI) upon infiltration of sulfate, either as FeSO4 or Na2SO4, was caused by anion exchange of sulfate for chromate in the layered double hydroxide mineral hydrocalumite, a process for which scanning electron microscopy with energy dispersive X-ray microanalysis provided direct evidence. PMID- 12901672 TI - Mechanistic studies of the photocatalytic oxidation of microcystin-LR: an investigation of byproducts of the decomposition process. AB - Microcystins (cyclic heptapeptides) are produced by a number of freshwater cyanobacteria and cause concern in potable water supplies due to their acute and chronic toxicity. The present study reports the structural characterization of the degradation products of the photocatalytic oxidation of microcystin-LR, so aiding the mechanistic understanding of this process. TiO2 photocatalysis is a promising technology for removal of these toxins from drinking water. However, before it can be adopted in any practical application it is necessary to have a sufficient knowledge of degradation byproducts and their potential toxicity. Liquid chromatography-mass spectrometry analysis demonstrated that the major destruction pathway of microcystin appears to be initiated via three mechanisms: UV irradiation, hydroxyl radical attack, and oxidation. UV irradiation caused geometrical isomerization of microcystin converting the (4E), (6E) of the Adda configuration to (4E), 6(Z) or 4(Z), 6(E). Hydroxyl radical attack on the conjugated diene structure of Adda moiety produced dihyroxylated products. Further oxidation cleaved the hydroxylated 4-5 and/or 6-7 bond of Adda to form aldehyde or ketone peptide residues, which then were oxidized into the corresponding carboxylic acids. Photocatalysis also hydrolyzed the peptide bond on the ring structure of microcystin to form linear structures although this appeared to be a minor pathway. PMID- 12901673 TI - Oxidation reaction of high molecular weight carboxylic acids in supercritical water. AB - Stearic acid, being a model compound of high molecular weight carboxylic acids, was oxidized in a batch reactor by changing the oxygen supply with an insufficient oxygen supply at a constant reaction time at 420 degrees C. On the basis of the intermediate products identified by GC/MS, NMR, and HPLC analyses and the free-radical reaction mechanism, the oxidation pathways of high molecular weight carboxylic acids in supercritical water are discussed. The reaction of carboxylic acids in supercritical water proceeds with the consecutive oxidation of higher molecular weight carboxylic acids to lower molecular weight carboxylic acids through several major pathways. The attack of the hydroxyl radical occurs not only at the carbons in alpha-, beta-, gamma-positions to a --COOH group but also at the carbons ((omega-1)-carbon and/or omega-carbon) far in the alkyl chain from a --COOH group, which may lead to the formation of dicarboxylic acids. PMID- 12901674 TI - Emissions of regulated pollutants from a spark ignition engine. Influence of fuel and air/fuel equivalence ratio. AB - A spark ignition engine is used to determine the influence of fuel composition and air/fuel equivalence ratio on the exhaust emissions of regulated pollutants. Two specific fuel matrices are used: the first contains eight hydrocarbons and the second contains four oxygenated compounds. A specific experimental design is used for these tests. Fuel aromatics increase the exhaust CO, HC, and NOx at stoichiometry, lean and rich conditions. Lambda is more important than fuel composition in the case of CO and HC. At stoichiometry, the addition of oxygenated compounds can decrease exhaust CO, HC, and NOx up to 30%, 50%, and 60%, respectively. Under these conditions, the addition of 5% of 2-propanol is the most effective for the reduction of CO, the addition of 20% of ethanol forthe reduction of HC, and this of 5% of methyl tributyl ester (MTBE) for the NOx. The addition of oxygenated compounds can decrease CO by 30% at lean conditions, while no decrease is observed at rich ones; HC and NOx can decrease up to 30% and 80%, respectively, under lean conditions and 50% under rich ones. At all lambda tested, exhaust NOx increases with the addition of 20% of 2-propanol. PMID- 12901675 TI - Comment on "Measurements of atmospheric mercury species at a coastal site in the Antarctic and over the South Atlantic Ocean during polar summer". PMID- 12901676 TI - The 1971 smallpox outbreak in the Soviet city of Aralsk: implications for Variola virus as a bioterrorist threat. Introduction. PMID- 12901677 TI - An epidemiological analysis of the 1971 smallpox outbreak in Aralsk, Kazakhstan. PMID- 12901678 TI - Report on measures taken to contain and eradicate the smallpox outbreak locale in the city of Aralsk (September/October, 1971). PMID- 12901679 TI - Report on measures taken to contain and eradicate the smallpox outbreak locale in the city of Aralsk, Part II. PMID- 12901680 TI - Commentary on implications of the 1971 outbreak in Aralsk, Kazakhstan, for U.S. Smallpox Vaccination Policy (No. 1). PMID- 12901681 TI - Commentary on the 1971 smallpox epidemic in Aralsk, Kazakhstan, and the Soviet Biological Warfare Program (No. 2). PMID- 12901682 TI - Commentary on Dr. Alan Zelicoff's epidemiological analysis of the Aralsk outbreak (No. 3). PMID- 12901683 TI - Commentary on Dr. Alan zelicoff's Analysis of the Aralsk outbreak (No. 4). PMID- 12901684 TI - Commentary on Dr. Alan P. Zelicoff's analysis (No. 5). PMID- 12901685 TI - Commentary on Dr. Alan P. Zelicoff's analysis (No. 6). PMID- 12901686 TI - Commentary on Dr. Alan P. Zelicoff's analysis (No. 7). PMID- 12901687 TI - A novel amylose corn-starch dispersion as an aqueous film coating for tablets. AB - A novel aqueous coating dispersion of amylose-rich corn starch (Hylon VII) was evaluated in an aqueous film-coating process of tablets using an instrumented laboratory-scale pan-coating apparatus. The influence of two independent process variables, the coating temperature and the atomizing air pressure, on the properties of the coated tablets were investigated. The preuse stability of aqueous coating dispersion (i.e., amylose corn-starch precipitate) was studied using a powder X-ray diffraction (XRD) technique. The crystallinity of amylose starch in the coating dispersion was found to increase slightly during 9 months of storage (in a refrigerator 6 +/- 2 degrees C). The film coatings of an aqueous amylose-rich starch dispersion were successfully applied onto tablets without any significant drawbacks, such as nozzle blockage or related problems. It was found that the temperature in the coating pan had a significant influence on the film surface roughness, mechanical strength, and drug release in vitro. When the lowest coating temperature (30 degrees C) was used, rougher film coatings were obtained due to overwetting. At higher temperatures (up to 50-60 degrees C), lower surface roughness and higher mechanical strength values for the coated tablets were obtained. With the present amylose starch dispersion, the atomizing air pressure had a minor influence on the quality of the coating. Under appropriate coating conditions, a smooth tablet film coating was produced with this new, natural, and inexpensive amylose starch dispersion. PMID- 12901688 TI - Effect of steam sterilization on the rheology of polymer solutions. AB - The effect of steam sterilization (dwell@121 degrees C) on the apparent viscosity of solutions of carbomer 940P, guar gum, hydroxyethylcellulose (HEC), and xanthan gum was studied. Guar gum and carbomer 940P could be steam sterilized for 30 min without any change in the apparent viscosity. Steam sterilization of HEC and xanthan gum resulted in a substantial decrease in the apparent viscosity and the desirable shear-thinning rheology. The addition of sodium chloride to either polymer prior to steam sterilization diminished the effect of sterilization on the rheology. The apparent viscosity of xanthan solutions was not significantly affected if the solution was made isotonic with sodium chloride (0.15 M). The addition of sodium chloride to xanthan gum after steam sterilization resulted in full recovery of the initial rheological properties. Carbomers, guar gum, and xanthan (in 0.15 M NaCI) are candidates for use in pharmaceutical solutions/suspensions that are subjected to steam sterilization. PMID- 12901689 TI - Effects of glycerol on the in vitro percutaneous absorption of all-trans retinoic acid. AB - The nature of the receptor solution plays an important role in in vitro percutaneous absorption of highly lipophilic compounds having limited solubility. In vitro permeation studies of a lipophilic compound, all-trans retinoic acid (RA), through the rat dorsal skin were performed with the presence of glycerol (0 20% v/v) in the receptor solution, and the results were compared with those with the presence of albumin (4%). The results showed that an addition of glycerol (20%) into the receptor solution significantly increased the permeation rate of RA through the rat dorsal skin (0.0068 +/- 0.0041 vs. 0.0014 +/- 0.0010 microg/cm2/hr). It was also found that RA tends to accumulate in the lipophilic layer, and its log P value between the epidermis and the receptor solution significantly decreased with the presence of glycerol (20%) (1.48 +/- 0.14 vs. 2.45 +/- 0.21). An addition of glycerol, an osmotherapeutic agent, in the physiological receptor solution seemed to enhance the percutaneous absorption of RA by affecting the partition coefficient of RA. PMID- 12901690 TI - Development and evaluation of a miniaturized procedure for determining the bonding index: a novel prototype for solid dosage formulation development. AB - The purpose of this study was a comparative evaluation of miniaturized vs. University of Minnesota's (i.e., U of Minn. = Hiestand's) procedures for determining the tensile strength, indentation hardness, and bonding index (BI), one out of three Indices of Tableting Performance (ITP). Tableting properties of six direct compression placebo formulations were determined by using a compaction simulator and a Texture Analyser TA-XT2I, or by following the U of Minn. method, which included a specially designed triaxial compression device, a computer controlled mechanical stress-strain analyzer, and a dynamic pendulum impact apparatus. Miniaturization of the procedures to determine the ITP, as well as the ability to differentiate between materials while operating compaction cycles more comparable to standard tablet production conditions, enabled proper evaluation of each material's inherent tableting properties. Indentation diameter calculated via an empirical equation appeared to correlate well with, and provided acceptable precision of accuracy to, the determination of indentation diameter via standard optical microscopy methods. The miniaturized and U of Minn. procedure exhibited a significant degree of correlation when comparing the BI. However, the tensile strength and indentation hardness values were somewhat different due to the use of triaxial decompression for the U of Minn. procedure vs. standard compaction profiling for the miniaturized procedure. The present direct compression placebo formulation data gathered from the miniaturized procedures and compared with the U of Minn. method for determining the ITP suggest that both techniques yield similar conclusions. However, discrimination of out-of-die compaction properties determined via the miniaturized procedures, such as tensile strength and indentation hardness, appeared to associate more precisely with changes in strain rate, thus allowing better discrimination of particle-particle interactions and ductile-to-brittle characteristics as a function of compaction speed and pressure. PMID- 12901691 TI - Stomach-specific anti-H. pylori therapy. II. Gastric residence studies of tetracycline-loaded chitosan microspheres in gerbils. AB - This study examines the gastric residence of chitosan microspheres and the local tetracycline concentrations following oral administration in gerbils. Chitosan microspheres were prepared by ionic cross-linking and precipitation with sodium sulfate. Gastric retention studies were performed by administering radioiodinated [125I] chitosan microsphere suspension in the nonacid-suppressed and acid suppressed states. At different time points, animals were sacrificed, and the radioactivity in tissues and fluids was measured with a gamma counter. Local tetracycline concentrations were measured using chitosan microspheres loaded with tritiated-[3H]-tetracycline. The radioactivity, measured with a liquid scintillation analyzer, was used to determine the microg of drug per gram of tissues or fluids under nonacid-suppressed and acid-suppressed states. Microspheres with a spherical shape and an average diameter of 2.0-3.0 microm were formed. After 2 hr in the fasted stomach, approximately 10% of the administered dose remained. The microspheres were predominantly found in the colon after 6 hr of administration. There was no detectable radioactivity in the small intestine, plasma, urine, liver, and kidneys. Additionally, acid suppression with ranitidine did not influence the gastric residence time of chitosan microspheres. To our disappointment, tetracycline concentration profile in the stomach from microsphere formulation was similar to the aqueous solution. Also, there was no significant difference between the stomach tetracycline concentrations in the nonacid-suppressed and acid-suppressed states. The drug was predominantly found in the colon and urine samples after 6 hr. Tetracycline could not be detected in the plasma, small intestine, liver, or kidneys. Results of this study show that chitosan microspheres prepared by ionic cross-linking do not provide a longer residence time in the fasted gerbil stomach. The tetracycline concentration profile in the stomach, following administration in microsphere formulation, was similar to that of aqueous solution. Lastly, acid suppression did not influence the gastric residence time of chitosan microspheres or tetracycline concentration profiles. PMID- 12901692 TI - Relationship between internal phase volume and emulsion stability: the cetyl alcohol/stearyl alcohol system. AB - The main objective of this study was to optimize the stability of cetyl alcohol/stearyl alcohol emulsions in terms of percentage of internal phase volume, emulsifier type and concentration, and amount of external phase (water). Creams (o/w emulsions) were prepared by phase inversion and physical properties as particle size of the internal phase, apparent viscosity, and sedimentation volume evaluated. Stability was performed at room temperature, 40 degrees C, 50 degrees C, and under stress conditions. High hydrophilic lipophilic balance (HLB) nonionic surfactants as tween 80, tween 20, Myrj 52, Brij 35, and low HLB span 60 were used as emulsifying agents. The percentage of internal phase components (cetyl alcohol and stearyl alcohol), percentage of emulsifying agents, and percentage of aqueous external phase were varied, and stability was investigated. As the level of emulsifier agent (tween 80 and span 60) increased from 3% to 15%, and the percent of the internal phase remained constant at 30%, the particle size of the internal phase decreased and the cream became more stable. Formulations of the same composition, but prepared using Myrj 53 and tween 20 as emulsifiers, showed a larger particle size than formulations prepared using tween 80 and Brij 35. As the level of the internal phase volume increased and consequently the amount of water decreased, emulsion viscosity increased. The best formulation containing 30% internal phase (50% cetyl alcohol, 35% stearyl alcohol), 15% emulsifying agents (tween 80/span 60 ratio of 3:1), and 70% water was selected, and effects of process temperature and cooling rate on emulsion stability investigated. This formulation was further investigated in terms of stability of a 1% hydrocortisone addition by varying the percentage (30%, 35%, 40%, and 45%) of internal phase and percentage of water (70%, 65%, 60%, and 55%). The best formulation contained 45% internal phase (22.5 g cetyl alcohol, 15.75 g stearyl alcohol, 15% emulsifying agent, which is equivalent to 5 g tween 80 and 1.7 g span 60), and 55% w/w water, was manufactured under different manufacturing processes. Emulsions prepared by homogenization at the beginning of the process of emulsification were stable with small internal phase particle diameter. This study demonstrates that at every cetyl alcohol/stearyl alcohol ratio there is a phase volume/emulsifier HLB ratio, which results in optimum stability. PMID- 12901693 TI - A study of the effects of curing and storage conditions on controlled release diphenhydramine HCl pellets coated with Eudragit NE30D. AB - The objective of this study was to investigate the possible impacts of curing and storage conditions on dissolution of controlled release diphenhydramine HCl pellets coated with EUDRAGIT NE30D. The accumulative percentage of dissolved active drug was used as the response in three statistical experimental design studies: 32 full factorial, Box-Behnken and 2(3) designs. By only considering curing temperature and curing time, both factors were found to significantly affect the dissolution rate, but curing temperature had greater impact than curing time. When considering polymer coating level, curing temperature and curing time together, polymer coating level and curing temperature had important effects on dissolution rate, but curing time became insignificant among these three factors. The addition of the water-soluble additives hydroxypropyl methyl cellulose and mannitol made coating films less sensitive to curing, and there was little or no difference in their effect in the model studied. Lower levels of a water-insoluble additive (kaolin) had little impact on dissolution; however, when the level of water-insoluble additive increased, the coating film became more sensitive to curing, especially at the lower curing temperature of 30 degrees C. PMID- 12901694 TI - Batch and continuous processing in the production of pharmaceutical granules. AB - A quasicontinuous granulation and drying process to avoid scale-up problems is introduced in this work. Consistent and reproducible granule quality is a key factor in robust dosage form design and fits ideally the prerequisites of a drug quality system for the twenty-first century and the Food and Drug Administration's Process Analytical Technology (PAT) initiative. In scale-up, factors that simulate or reproduce the laboratory scale must be considered. This system provides a new possibility for industrial manufacturing and galenical development of pharmaceutical solids. The quasicontinuous method described in the present work, and the laboratory and production batches and the granulating equipment used to produce them, are the same. Once a robust process has been defined in the laboratory, it is merely repeated as many times as necessary to achieve the desired final batch size. The quasicontinuous process gives new possibilities to simplify manufacturing procedures and to validate them faster. The quality of the resulting granules and tablets compared with classical methods is equal until better. In many cases, existing products have been transferred to the multicell process without formulation changes. The quasicontinuous production concept for high-shear granulation and fluid-bed drying offers many advantages over the classical methods used to produce pharmaceutical granules. The wet massing process may be monitored by the power consumption of the mixer motor for each subunit, as in classical high-shear granulation processes. The air volume, temperature, and humidity of each of the drying cells may be controlled individually to avoid overheating of temperature-sensitive materials. All processing variables must be precisely controlled by a computer, and the data must be collected for documentation. As such, product quality and reproducibility for each subunit is assured. PMID- 12901695 TI - Scale-up and optimization of an evaporative drying process applied to aqueous dispersions of solid lipid nanoparticles. AB - An apparatus to dry aqueous dispersions of solid lipid nanoparticles (SLNs) was designed. Optimal running conditions were evaluated to obtain minimum process time and produce dried SLNs characterized by small size variation. To achieve process optimization, SLN average diameter, SLNs polydispersity index, and drying time were related to three operative variables: process temperature, SLN concentration in the original aqueous dispersions, and nitrogen flow rate as the physical means of the drying process. An experimental design procedure and a multicriteria optimization method, targeting desirability functions, enabled us to obtain the optimal conditions for all responses. Drying time, average diameter, and polydispersity index of dried SLN batches were more favorable than those obtained by freeze-drying identical SLN aqueous dispersions with the same initial nanoparticle concentration. PMID- 12901696 TI - Composite film-coated tablets intended for colon-specific delivery of 5 aminosalicylic acid: using deesterified pectin. AB - Combinations of Eudragit RS and deesterified pectin, polygalacturonic acid (PGA), or its potassium and sodium salts, when applied as a film coat, has a potential value as a colon-specific delivery system. Dispersions of PGA in Eudragit RS were used as the film former for coating of 5-aminosalicylic acid (5-ASA) tablet cores. Drug release behavior was assessed, in vitro, under simulating conditions in term of pH and time to in vivo during their transit to the colon. Negligible drug release occurred during first 5 hr where the coated tablets were in the stomach and small intestine. After that, the pectinolytic enzymes were added into the pH 6.8 medium to simulate the in vivo condition where there is the digestion of bacteria in the colon. The release of 5-ASA from the coated tablets occurred linearly as a function of time. Drug release depended on the composition of the mixed film, as well as the ratio of Eudragit RS to PGA or its salts. The highest drug release from the coated tablets of about 40% was obtained when the ratio of Eudragit RS to potassium salt of PGA was 2.5 to 1. Drug release profiles seemed to conform to the mechanism involving the osmotically driven release and formation of channels in the film caused by dissolution of PGA salts. Channel formation was, in most cases, activated by the presence of pectinolytic enzymes, showing that the PGA in the mixed film was subjected to enzymic breakdown. In conclusion, PGA could be used as an additive in Eudragit RS films to control the release of colonic delivery system. PMID- 12901697 TI - Dominantly inherited peripheral neuropathies. AB - Since 1886, the year that Charcot and Marie and Tooth described a genetic "peroneal muscular atrophy syndrome," electrophysiological and histological studies of the peripheral nervous system have greatly aided the characterization of this syndrome, which falls among the hereditary sensory-motor neuropathies. Two principal forms of Charcot-Marie-Tooth (CMT) disease have been distinguished: CMT 1, corresponding to a demyelinating type, and CMT 2, corresponding to an axonal type. The modes of transmission of these types are variable, recessive or dominant, autosomal, or X-linked. Our discussion here is confined to the dominant forms. In recent years, advances in molecular biology have greatly modified the approach to CMT disease and related neuropathies (such as hereditary neuropathy with liability to pressure palsies). With increased knowledge of responsible gene mutations and several other loci identified by linkage studies, our understanding of the pathophysiology of these neuropathies is increasing; however, with greater understanding, the classification of these disorders is becoming more complex. In this review we present and discuss the currently characterized subtypes, with emphasis on their known histological aspects. While nerve biopsy has lost its diagnostic importance in certain forms of the disease, such as CMT 1A, CMT 1B, and X-linked CMT (CMT X), it remains important for better characterizing the lesions of CMT 2 and forms of genetic peroneal atrophy in which DNA study is unrevealing. PMID- 12901698 TI - Brain banking in the United States. AB - Many of us who conduct research based on disorders that uniquely affect the human nervous system are involved directly or indirectly with brain banks. Brain banking is by its very nature a multi-disciplinary endeavor that requires close collaboration with our colleagues in clinical departments and also with the families and patients who are donors of brain tissue. These brain tissues will ultimately be used for many types of basic science investigations. Thus, many brain banks are as unique as the basic research programs that they serve. However, there are some commonalities in the field of brain banking. This paper represents a broad overview of brain banking and the issues that are common to all brain banks. Legal and ethical concerns regarding confidentiality of donor records and donor recruitment procedures, as well as safety precautions for technical staff, tissue banking methods and disbursement will be discussed. Finally, issues surrounding financial support for brain banks will be considered. PMID- 12901699 TI - Subpial demyelination in the cerebral cortex of multiple sclerosis patients. AB - The extent and pattern of demyelination in the cerebral cortex was determined in 78 tissue blocks from the brains of 20 multiple sclerosis (MS) patients and 28 tissue blocks from 7 patients without neurological disease. Tissue blocks from 4 predetermined areas (cingulate gyrus, frontal, parietal, and temporal lobe) were studied, irrespective of macroscopically evident MS plaques. All tissue blocks contained cerebral cortex and periventricular and/or subcortical white matter. One hundred and nine demyelinating lesions were detected in the cerebral cortex, of which 92 (84.4%) were purely intracortical and 17 (15.6%) were lesions extending through both white and gray matter areas. In 5 of the 20 MS brains, subpial demyelination was extensive in the 4 widely spaced cortical areas studied, thus considered to represent a general cortical subpial demyelination. The percentage of demyelinated area was significantly higher in the cerebral cortex (mean 26.5%, median 14.1%) than in white matter (mean 6.5%, median 0%) (p = 0.001). Both gray and white matter demyelination was more prominent in the cingulate gyrus than in the other areas examined (p < 0.05). These results indicate that the cerebral cortex is likely to be a predilection site for MS lesions and identify general cortical subpial demyelination as a distinct pattern occurring in a significant subpopulation of MS patients. PMID- 12901700 TI - GluR2(B) knockdown accelerates CA3 injury after kainate seizures. AB - Ca2+ currents are thought to enhance glutamate excitotoxicity. To investigate whether reduced expression of the Ca2+ limiting GluR2(B) subunit enhances seizure induced vulnerability to either CA1 or CA3 neurons, we delivered GluR2(B) oligodeoxynucleotides (AS-ODNs) to the dorsal hippocampus of adult rats before inducing kainate (KA) seizures. After knockdown, no changes in behavior, electrographic activity, or histology were observed. In contrast, GluR2(B) knockdown and KA-induced status epilepticus produced accelerated histological injury to the ipsilateral CA3a-b and hilar subregions. At 8 to 12 h, the CA3a was preferentially labeled by both silver and TUNEL methods. TUNEL staining revealed 2 types of nuclei. They were round with uniform label, features of necrosis, or had DNA clumping or speckled chromatin deposits within surrounding cytosol, features of apoptosis. At 16 to 24 h, many CA3a-c neurons were shrunken, eosinophilic, argyrophilic, or completely absent. Immunohistochemistry revealed marked decreases in GluR2(B) subunits throughout the hippocampus, NR1 immunoreactivity was also reduced but to a lesser extent. In contrast, GluR1 and NR2A/B immunohistochemistry was relatively uniform except in regions of cell loss or within close proximity to the CA1 infusion site. At 144 h, the CA3 was still preferentially injured although bilateral CA1 injury was also observed in some AS ODN-, S-ODN-, and KA-only-treated animals. Glutamate receptor antibodies revealed generalized decreases in the CA3 with all probes tested at this delayed time. In contrast, GluR2(B) expression was increased within CA1 irregularly shaped, injured neurons. Therefore, hippocampal deprivation of GluR2(B) subunits is insufficient to induce cell death in mature animals but may accelerate the already known CA3/hilar lesion, possibly by triggering apoptosis within CA3 neurons. CA1 and DG survive the first week despite their loss of GluR2(B) subunits, suggesting that other intrinsic properties such as increased Na+ conductance and reduced ability of the GluR2(B) subunit to interact with certain cytoplasmic proteins may be responsible for the augmented cell death rather than changes in AMPA receptor-mediated Ca2+ permeability. Alternatively, changes in allosteric interactions that affect other receptor classes of high density at the mossy fiber synapse (e.g. KA receptors) may augment KA neurotoxicity. Latent GluR2(B) increases in CA1 injured neurons support a role for AMPA receptor subunit alterations in seizure-induced tolerance. PMID- 12901701 TI - Phenotypic differences between peripheral myelin protein-22 (PMP22) and myelin protein zero (P0) mutations associated with Charcot-Marie-Tooth-related diseases. AB - Mutations in the genes for peripheral myelin protein-22 (PMP22) and myelin protein zero (P0) cause human hereditary neuropathies with varying clinical and pathological phenotypes. In this study, we examine the effects of representative disease-causing mutations on the subcellular distribution of their corresponding PMP22- and P0-enhanced green fluorescent protein (EGFP) fusion proteins. In transiently transfected HeLa and 293 cells, we find that wild-type P0-EGFP and PMP22-EGFP are efficiently synthesized and transported through the secretory pathway to the plasma membrane. The P0-EGFP and PMP22-EGFP mutants can be classified into several groups: those that are transported to the plasma membrane as in the majority of P0 mutants; those that are retained in the endoplasmic reticulum as in the majority of PMP22 mutants; and those that are a mixture of the two. In addition, several of these disease-causing mutations are associated with the development of abnormal intracellular cytoplasmic structures that we have previously identified as either intracellular myelin figures or aggresomes. Our studies indicate that different types of PMP22 and P0 mutations are associated with specific intracellular chaperone proteins, including calnexin and BiP, and that these associations can be altered by glycosylation. These findings indicate that the various P0 and PMP22 mutants may exert their pathogenic effects in different subcellular compartments and by different mechanisms in the mammalian cell. PMID- 12901702 TI - Proinsulin C-peptide replacement in type 1 diabetic BB/Wor-rats prevents deficits in nerve fiber regeneration. AB - We recently reported that early gene responses and expression of cytoskeletal proteins are perturbed in regenerating nerve in type 1 insulinopenic diabetes but not in type 2 hyperinsulinemic diabetes. We hypothesized that these differences were due to impaired insulin action in the former type of diabetes. To test this hypothesis, type 1 diabetic BB/Wor-rats were replaced with proinsulin C-peptide, which enhances insulin signaling without lowering blood glucose. Following sciatic nerve crush injury, early gene responses such as insulin-like growth factor, c-fos, and nerve growth factor were examined longitudinally in sciatic nerve. Neurotrophic factors, their receptors, and beta-tubulin and neurofilament expression were examined in dorsal root ganglia. C-peptide replacement significantly normalized early gene responses in injured sciatic nerve and partially corrected the expression of endogenous neurotrophic factors and their receptors, as well as neuroskeletal protein in dorsal root ganglia. These effects translated into normalization of axonal radial growth and significantly improved axonal elongation of regenerating fibers in C-peptide-replaced BB/Wor-rats. The findings in C-peptide replaced type 1 diabetic rats were similar to those previously reported in hyperinsulinemic and iso-hyperglycemic type 2 BB/Z-rats. We conclude that impaired insulin action may be more important than hyperglycemia in suppressing nerve fiber regeneration in type 1 diabetic neuropathy. PMID- 12901703 TI - Heparan sulfate proteoglycans modulate monocyte migration across cerebral endothelium. AB - Heparan sulfate proteoglycans (HSPGs) are known to participate in a wide range of biological events, including cellular trafficking. In this study we report that in situ cerebral blood vessels highly express HSPGs. Of the syndecan family, syndecan-2 is highly expressed on virtually all brain vessels and syndecan-1 and 3 are only present on larger blood vessels. These endothelial HSPGs have a functional role in monocyte diapedesis across brain endothelium, as assessed in our in vitro adhesion and migration assays. Our data indicate that heparin prevents monocyte adhesion to brain endothelium by interacting solely with the monocyte. Transendothelial migration of monocytes can be prevented by preincubation of brain endothelium with heparin by enzymatic removal of heparan sulphate side chains or by inhibition of cellular sulfation. Blocking of G protein-dependent signaling in the monocytes prevented monocyte adhesion and migration to similar extent, suggesting that G-dependent signaling may be involved in HSPG-mediated monocyte adhesion and transendothelial migration. Our data demonstrate that brain endothelial HSPGs have a modulatory role in the transendothelial migration of monocytes in a direct and indirect fashion and may therefore contribute to the formation of neuroinflammatory lesions. PMID- 12901704 TI - Biochemical and ultrastructural study of neurofibrillary tangles in amyotrophic lateral sclerosis/parkinsonism-dementia complex in the Kii peninsula of Japan. AB - Amyotrophic lateral sclerosis/parkinsonism-dementia complex of the Kii peninsula (Kii ALS/PDC) is a neurodegenerative disorder endemic to natives in the southern coast area of the Kii peninsula of Japan. The disorder closely resembles Guamanian ALS/PDC clinically and neuropathologically. The characteristic neuropathological finding is abundant neurofibrillary tangles (NFTs) without amyloid deposition. To elucidate the biochemical properties of hyperphosphorylated tau protein, the major component of the NFTs, we examined Kii ALS/PDC brains by immunoblotting and immunohistochemical analysis using well characterized anti-tau antibodies specific to phosphorylation-dependent or independent epitopes. Hyperphosphorylated tau in Kii ALS/PDC had phosphorylated epitopes common to tau of paired helical filaments (PHFs) in Alzheimer disease (AD): immunoblot showed triplet bands composed of 6 tau isoforms. Ultrastructurally, NFTs revealed a twisted filamentous shape similar to PHF of AD. The biochemical properties of its phosphorylated tau protein and the ultrastructural characteristics of the NFTs of Kii ALS/PDC are very similar, if not identical, to PHF tau in AD, although they are different taupopathies. PMID- 12901705 TI - Prevalence of epilepsy and associated health service utilization and mortality among patients with intellectual disability. AB - We considered the prevalence of epilepsy and associated health service utilization for a population with intellectual disability. Registers for epilepsy and intellectual disability were created using a range of datasets. Of 1,595 people with an intellectual disability, 257 (16.1%) had epilepsy. Standardized activity ratios were 3.07 (95% CI 3.00 to 3.15), 2.03 (95% CI 1.94 to 2.11), and 3.09 (95% CI 2.78 to 3.41) for inpatients, outpatients, and accident and emergency, respectively. After excluding epilepsy-related inpatient admissions, we found the standardized activity ratio was 2.55 (2.48 to 2.62). Institutionalized patients were less likely to be admitted than were those in the community (standardized activity ratio = 0.63 (95% CI 0.54 to 0.73). Patients with intellectual disability and co-existing epilepsy used secondary care services more frequently than did those with intellectual disability only. PMID- 12901706 TI - Sequential analysis reveals a unique structure for self-injurious behavior. AB - Conditional probability, calculated using sequential analysis techniques in four time conditions (2, 10, 30, and 60 seconds), provided evidence that successive episodes of self-injury were sequentially dependent. This unique distribution of sequential association for self-injurious behavior (SIB) was not related to frequency or rate of occurrence. Compared with other environmental and behavioral events, the best predictor of SIB was an earlier SIB episode, consistent with a contagious distribution. This study is one of the few in which sequential analysis techniques were applied to data derived from a large group of individuals with severe behavior disorders. It may be the first in which this analytic tool was used to investigate systematically successive occurrences of SIB as it takes place in vivo. PMID- 12901707 TI - Behavioral relationship between autism and fragile x syndrome. AB - Previous researchers have reported autistic features in children with fragile X syndrome. We compared 21 children with pervasive developmental disorders (autism group) to 15 with fragile X syndrome on the Childhood Autism Rating Scale and the Reiss Scales for Children's Dual Diagnosis. The 7 children (47%) with fragile X who scored above the Childhood Autism Rating Scale cut-off (fragile X-autism group) were more impaired than the remaining children (fragile X-no autism) on Childhood Autism Rating Scale subscales related to emotion, visual and listening responses, and communication. The autism group's Reiss scores were higher than fragile X-no autism group, but not fragile X-autism group. Although the Childhood Autism Rating Scale identified almost 50% of children with fragile X as having autism, qualitative differences may exist in specific autistic-like behaviors between children with autism and children with fragile X. PMID- 12901708 TI - Maladaptive behaviors related to adaptive decline in aging adults with mental retardation. AB - Changes in patterns of maladaptive behavior related to age-associated adaptive declines were investigated in 529 adults with mental retardation. Although individuals with no significant adaptive decline displayed stable patterns of maladaptive behavior over a 3-year period, those with declines in function showed more variable patterns. Certain maladaptive behaviors were related to the onset of adaptive declines, with some becoming of increasing concern even before adaptive declines were noted (e.g., lack of boundaries). Other behaviors increased as adaptive declines developed (e.g., withdrawal). In general, findings suggest similarities in the course of age-associated dementia of adults with and without mental retardation and indicate that increases in selected areas of maladaptive behavior may be early indicators of concern for individuals at risk. PMID- 12901709 TI - Appendicular fractures: a significant problem among institutionalized adults with developmental disabilities. AB - A high incidence of nontraumatic fracture in adults with developmental disabilities living in a state-run facility was described. Risk factors for fracture, including bone mineral density (BMD), were investigated to determine whether people at highest risk for fracture could be prospectively identified. There was a 7.3% incidence of fracture among 391 adults. Risk factors were examined for 23 residents with fracture and 23 age-, race-, and gender-matched controls. There was a trend for antiepileptic medication usage to be associated with fractures. Estimated BMD by heel ultrasound did not predict fracture; however, values were much lower than those for the general population. Fractures and low BMD are significant problems among institutionalized adults with severe developmental disabilities. Further studies to identify therapies to prevent fractures are warranted. PMID- 12901710 TI - Subtypes of nonsocial play: comparisons between young children with and without developmental delays. AB - Subtypes of nonsocial play were examined for matched groups of young typically developing children and children with mild developmental (cognitive) delays. Based on intercorrelation patterns, interindividual stability, and associations with peer interaction measures, findings indicated that the nonsocial play of children with delays can be characterized as multidimensional in a manner similar to that of typically developing children. However, analyses suggest different interpretations of the psychological meaning of nonsocial play subtypes for both groups of children than indicated by previous work. Context factors did not influence nonsocial play subtypes. Evidence was found indicating considerable heterogeneity within subtypes, suggesting the need for longitudinal studies focusing on children's peer-related social competence, which may reveal subgroups within subtypes that both refine the constructs and provide direction for clinical interventions. PMID- 12901711 TI - Flicker flutter: is an illusory event as good as the real thing? AB - Verghese and Stone (1995) showed that reducing the perceived number of objects by grouping also reduces objective performance. Shams, Kamitani, and Shimojo (2000) showed that a single flash accompanied by multiple beeps appears to flash more than once. We show that objective orientation-discrimination performance depends solely on the perceived number of flashes, independent of the actual number of beeps and flashes. Thus the unit of perceptual analysis seems to be a perceived event, independent of how it is induced. PMID- 12901712 TI - Bootstrapped learning of novel objects. AB - Recognition of familiar objects in cluttered backgrounds is a challenging computational problem. Camouflage provides a particularly striking case, where an object is difficult to detect, recognize, and segment even when in "plain view." Current computational approaches combine low-level features with high-level models to recognize objects. But what if the object is unfamiliar? A novel camouflaged object poses a paradox: A visual system would seem to require a model of an object's shape in order to detect, recognize, and segment it when camouflaged. But, how is the visual system to build such a model of the object without easily segmentable samples? One possibility is that learning to identify and segment is opportunistic in the sense that learning of novel objects takes place only when distinctive clues permit object segmentation from background, such as when target color or motion enables segmentation on single presentations. We tested this idea and discovered that, on the contrary, human observers can learn to identify and segment a novel target shape, even when for any given training image the target object is camouflaged. Further, perfect recognition can be achieved without accurate segmentation. We call the ability to build a shape model from high-ambiguity presentations bootstrapped learning. PMID- 12901713 TI - Temporal dynamics of early light adaptation. AB - This study investigated two aspects of visual sensitivity to a change in light level. The first experiment examined the time course of sensitivity to a small amplitude 1-s contrast pedestal presented on a 163-td pedestal within a 115-td surround. The largest contrast pedestal was an 8% contrast change that changed the steady pedestal threshold by only 0.03 log unit. Thresholds increased by 0.6 log unit or more at both onset and offset, with a return to baseline within 100 ms. The increment and decrement thresholds showed different time courses. Increment thresholds were raised before pedestal onset and decrement thresholds were raised before pedestal offset. These data were interpreted to show that increment thresholds measured at onset and offset of a contrast pedestal accessed on-pathways and decrement thresholds accessed off-pathways. In the second experiment, we examined the time course of sensitivity using a Crawford paradigm. Observers were dark-adapted and sensitivity was measured before, during, and after a 0.5-s, 68-td pulse. Test stimuli were slightly biased in chromaticity and the observer reported the presence and the hue appearance of the test, allowing separate estimates of detection and hue identification. Thresholds increased during the pulse but showed only a minimal overshoot. The test sensitivity for detection was poorer than for hue identification in the dark, but detection and identification were similar during the pulse. These data suggest that thresholds during sizable luminance pulses are determined in the PC-pathway. PMID- 12901714 TI - Pattern-onset stimulation boosts central multifocal VEP responses. AB - Multifocal visual evoked potentials (VEP) allow one to assess whether stimulation at specific visual field locations elicits cortical activity; it might therefore enable us to conduct objective visual field perimetry. However, due to the cortical folding, which differs markedly between subjects, a particular electroencephalogram generator may fail to project signal on some recording electrodes. This may lead to false alarms for potential scotomata. Here we compare pattern-reversal and pattern-onset stimulation in their efficacy to activate the visual cortex and recorded mfVEPs to 60 locations comprising a visual field of 44 degrees diameter. We report three main findings: (1) Pattern onset compared to pattern-reversal enhances the amplitude by 30% for stimulation of the central visual field (<10 degrees radius), while evoking 30% less response in the periphery (>15 degrees ). (2) Although pattern-onset and pattern-reversal responses differ markedly in their eccentricity dependence, they have a similar topographical distribution. (3) By combining both stimuli, the number of false positives was reduced to less than 1.5% of the visual field locations tested. We conclude that pattern-onset and pattern-reversal activate identical visual cortical areas but target different neural mechanisms within these areas. Furthermore, pattern-onset stimulation greatly increases the sensitivity of the mfVEP to assess the cortical representation of the central 10 degrees of the visual field. PMID- 12901715 TI - Is it an animal? Is it a human face? Fast processing in upright and inverted natural scenes. AB - Object categorization can be extremely fast. But among all objects, human faces might hold a special status that could depend on a specialized module. Visual processing could thus be faster for faces than for any other kind of object. Moreover, because face processing might rely on facial configuration, it could be more disrupted by stimulus inversion. Here we report two experiments that compared the rapid categorization of human faces and animals or animal faces in the context of upright and inverted natural scenes. In Experiment 1, the natural scenes contained human faces and animals in a full range of scales from close-up to far views. In Experiment 2, targets were restricted to close-ups of human faces and animal faces. Both experiments revealed the remarkable object processing efficiency of our visual system and further showed (1) virtually no advantage for faces over animals; (2) very little performance impairment with inversion; and (3) greater sensitivity of faces to inversion. These results are interpreted within the framework of a unique system for object processing in the ventral pathway. In this system, evidence would accumulate very quickly and efficiently to categorize visual objects, without involving a face module or a mental rotation mechanism. It is further suggested that rapid object categorization in natural scenes might not rely on high-level features but rather on features of intermediate complexity. PMID- 12901716 TI - Characterization of the lipid-binding domain of the Plasmodium falciparum CTP:phosphocholine cytidylyltransferase through synthetic-peptide studies. AB - Phospholipid biosynthesis plays a key role in malarial infection and is regulated by CCT (CTP:phosphocholine cytidylyltransferase). This enzyme belongs to the group of amphitropic proteins which are regulated by reversible membrane interaction. To assess the role of the putative membrane-binding domain of Plasmodium falciparum CCT (PfCCT), we synthesized three peptides, K21, V20 and K54 corresponding to residues 274-294, 308-327 and 274-327 of PfCCT respectively. Conformational behaviour of the peptides, their ability to bind to liposomes and to destabilize lipid bilayers, and their insertion properties were investigated by different biophysical techniques. The intercalation mechanisms of the peptides were refined further by using surface-pressure measurements on various monolayers at the air/water interface. In the present study, we show that the three studied peptides are able to bind to anionic and neutral phospholipids, and that they present an alpha-helical conformation upon lipid binding. Peptides V20 and the full-length K54 intercalate their hydrophobic parts into an anionic bilayer and, to a lesser extent, a neutral one for V20. Peptide K21 interacts only superficially with both types of phospholipid vesicles. Adsorption experiments performed at the air/water interface revealed that peptide K54 is strongly surface-active in the absence of lipid. Peptide V20 presents an atypical behaviour in the presence of phosphatidylserine. Whatever the initial surface pressure of a phosphatidylserine film, peptide V20 and phosphatidylserine entities seem linked together in a special organization involving electrostatic and hydrophobic interactions. We showed that PfCCT presents different lipid dependence properties from other studied CCTs. Although the lipid-binding domain seems to be located in the C-terminal region of the enzyme, as with the mammalian counterpart, the membrane anchorage, which plays a key role in the enzyme regulation, is driven by two alpha-helices, which behave differently from one another. PMID- 12901717 TI - Eukaryotic translation-initiation factor eIF2beta binds to protein kinase CK2: effects on CK2alpha activity. AB - eIF2 (eukaryotic translation-initiation factor 2) is a substrate and an interacting partner for CK2 (protein kinase CK2). Co-immuno-precipitation of CK2 with eIF2beta has now been observed in HeLa cells, overexpressing haemagglutinin tagged human recombinant eIF2beta. A direct association between His6-tagged human recombinant forms of eIF2beta subunit and both the catalytic (CK2alpha) and the regulatory (CK2beta) subunits of CK2 has also been shown by using different techniques. Surface plasmon resonance analysis indicated a high affinity in the interaction between eIF2beta and CK2alpha, whereas the affinity for the association with CK2beta is much lower. Free CK2alpha is unable to phosphorylate eIF2beta, whereas up to 1.2 mol of phosphate/mol of eIF2beta was incorporated by the reconstituted CK2 holoenzyme. The N-terminal third part of eIF2beta is dispensable for binding to either CK2alpha or CK2beta, although it contains the phosphorylation sites for CK2. The remaining central/C-terminal part of eIF2beta is not phosphorylated by CK2, but is sufficient for binding to both CK2 subunits. The presence of eIF2beta inhibited CK2alpha activity on calmodulin and beta casein, but it had a minor effect on that of the reconstituted CK2 holoenzyme. The truncated forms corresponding to the N-terminal or central/C-terminal regions of eIF2beta were much less inhibitory than the intact subunit. The results demonstrate that the ability to associate with CK2 subunits and to serve as a CK2 substrate are confined to different regions in eIF2beta and that it may act as an inhibitor on CK2alpha. PMID- 12901718 TI - Trafficking of Kv1.4 potassium channels: interdependence of a pore region determinant and a cytoplasmic C-terminal VXXSL determinant in regulating cell surface trafficking. AB - Kv1.4 and Kv1.1 potassium channel homomers have been shown to exhibit different intracellular trafficking programmes and cell-surface expression levels in cell lines: a determinant in the pore region of Kv1.4 and Kv1.1 [Zhu, Watanabe, Gomez and Thornhill (2001) J. Biol. Chem. 276, 39419-39427] and a cytoplasmic C terminal VXXSL determinant on Kv1.4 [Li, Takimoto and Levitan (2000) J. Biol. Chem. 275, 11597-11602] have been described, which affected trafficking and cell surface expression levels. In the present study, we examined whether trafficking pore determinants influenced any cytoplasmic C-terminal trafficking determinant. We found that removal of VXXSL from a Kv1.4 chimaera that contained the pore of Kv1.1 did not affect cell-surface trafficking. Therefore removal of the C terminal VXXSL of Kv1.4 inhibited protein surface levels only in the presence of the Kv1.4 pore. In contrast, truncating the cytoplasmic C-terminus of Kv1.1 or truncating a Kv1.1 chimaera with the pore of Kv1.4, had little effect on surface protein levels. Furthermore, the subregion of the Kv1.4 pore trafficking determinant that was required for the inhibitory effect of VXXSL removal was mapped to a threonine residue in the deep pore region. Therefore the Kv1.4 pore determinant affected the trafficking and cell-surface levels directed by the C terminal VXXSL determinant. Different Kv1 trafficking programmes would affect cell-surface expression levels either positively or negatively and also cell signalling. Cells may use differential trafficking programmes of membrane proteins as a post-translational mechanism to regulate surface protein levels and cell function. PMID- 12901719 TI - Regulation of oxidative phosphorylation in different muscles and various experimental conditions. AB - It has been shown previously that direct stimulation of oxidative-phosphorylation complexes in parallel with the stimulation of ATP usage is able to explain the stability of intermediate metabolite (ATP/ADP, phosphocreatine/creatine, NADH/NAD+, protonmotive force) concentrations accompanied by a large increase in oxygen consumption and ATP turnover during transition from rest to intensive exercise in skeletal muscle. It has been also postulated that intensification of parallel activation in the ATP supply-demand system is one of the mechanisms of training-induced adaptation of oxidative phosphorylation in skeletal muscle. In the present paper, it is demonstrated, using the computer model of oxidative phosphorylation in intact skeletal muscle developed previously, that the direct activation of oxidative phosphorylation during muscle contraction can account for the following kinetic properties of oxidative phosphorylation in skeletal muscle encountered in different experimental studies: (i) increase in the respiration rate per mg of mitochondrial protein at a given ADP concentration as a result of muscle training and decrease in this parameter in hypothyroidism; (ii) asymmetry (different half-transition time, t(1/2)) in phosphocreatine concentration time course between on-transient (rest-->work transition) and off-transient (recovery after exercise); (iii) overshoot in phosphocreatine concentration during recovery after exercise; (iv) variability in the kinetic properties of oxidative phosphorylation in different kinds of muscle under different experimental conditions. No other postulated mechanism is able to explain all these phenomena at the same time and therefore the present paper strongly supports the idea of the parallel activation of ATP usage and different oxidative-phosphorylation complexes during muscle contraction. PMID- 12901720 TI - Variation of the axial haem ligands and haem-binding motif as a probe of the Escherichia coli c-type cytochrome maturation (Ccm) system. AB - Cytochromes c are typically characterized by the covalent attachment of haem to polypeptide through two thioether bonds with the cysteine residues of a Cys-Xaa Xaa-Cys-His peptide motif. In many Gram-negative bacteria, the haem is attached to the polypeptide by the periplasmically functioning cytochrome c maturation (Ccm) proteins. Exceptionally, Hydrogenobacter thermophilus cytochrome c552 can be expressed as a stable holocytochrome both in the cytoplasm of Escherichia coli in an apparently uncatalysed reaction and also in the periplasm in a Ccm-mediated reaction. In the present study we show that a Met60-->Ala variant of c552, which does not have the usual distal methionine ligand to the haem iron of the mature cytochrome, can be made in the periplasm by the Ccm system. However, no holocytochrome could be detected when this variant was expressed cytoplasmically. These data highlight differences between the two modes of cytochrome c assembly. In addition, we report investigations of haem attachment to cytochromes altered to have the special Cys-Trp-Ser-Cys-Lys haem-binding motif, and Cys-Trp-Ser-Cys His and Cys-Trp-Ala-Cys-His analogues, of the active-site haem of nitrite reductase NrfA. PMID- 12901722 TI - Shear-induced release of disabled herpes simplex virus from baby-hamster kidney cells. AB - A new process route is proposed to increase the production yield of disabled herpes simplex virus type 1 (HSV-1 DIS). Infected baby-hamster kidney (BHK) cells were subjected to a range of shear rates between 3.69 x 10(3) s(-1) and 51.3 x 10(3) s(-1) in the gap between a pair of co-axial cylinders. Analysis of the supernatant fractions of sheared material established that optimal virus release was achieved by exposing the infected cells to a shear rate of 42.7 x 10(3) s(-1) for a period of 1 min. Compared with the current laboratory process, the titre of HSV-1 DIS was increased over 30-fold, from about 1 x 10(6) to 30 x 10(6) pfu (plaque-forming units)/ml. Evaluation of the supernatant fractions by flow cytometry, total protein assay, PAGE and dot-blot assays showed no evidence of cell disruption, supporting the hypothesis that shear-induced release of the cell membrane-bound virus was achieved without compromising downstream purification. The proposed method is scalable, and since no additional chemicals are required, it provides an attractive option for enhanced recovery of virus particles for therapeutic applications. PMID- 12901721 TI - Fine mapping of sequential neutralization epitopes on the subunit protein VP8 of human rotavirus. AB - The epitopes of the HRV (human rotavirus), especially those involved in virus neutralization, have not been determined in their entirety, and would have significant implications for HRV vaccine development. In the present study, we report on the epitope mapping and identification of sequential neutralization epitopes, on the Wa strain HRV subunit protein VP8, using synthetic overlapping peptides. Polyclonal antibodies against recombinant Wa VP8 were produced previously in chicken, and purified from egg yolk, which showed neutralizing activity against HRV in vitro. Overlapping VP8 peptide fragments were synthesized and probed with the anti-VP8 antibodies, revealing five sequential epitopes on VP8. Further analysis suggested that three of the five epitopes detected, M1-L10, I55-D66 and L223-P234, were involved in virus neutralization, indicating that sequential epitopes may also be important for the HRV neutralization. The interactions of the antibodies with the five epitopes were characterized by an examination of the critical amino acids involved in antibody binding. Epitopes comprised primarily of hydrophobic amino acid residues, followed by polar and charged residues. The more critical amino acids appeared to be located near the centre of the epitopes, with proline, isoleucine, serine, glutamine and arginine playing an important role in the binding of antibody to the VP8 epitopes. PMID- 12901724 TI - Multiple primary tumours following head and neck cancer in southern England during 1961-98. AB - BACKGROUND: Successful initial treatment of oral and oro-phayngeal cancer has led to the emergence of second primary tumours (SPTs). Population data are meagre. METHODS: Occurrence of multiple primary cancers following a malignancy in a head and neck site was computed using data from a population-based cancer registry covering a population of 14 million. RESULTS: Among 59,958 subjects reported to the registry, 5.5% males and 3.6% females developed a second primary cancer. At the sites studied, a total of 2771 second primary cancers were found, compared with an expected number of 2341. The standardised incidence ratio (SIR) for contracting a new primary cancer was 1.14 (95% CI=1.09-1.19) for men and 1.34 (95% CI=1.24-1.44) for women. There was a significantly increased risk for a second cancer in most of the upper aerodigestive tract sites that are generally regarded as tobacco associated, with an SIR for subsequent oral cancer of 5.56 in men and 15.31 in women. Subjects first detected with a pharyngeal cancer experienced the highest SIR for a subsequent tumour. Excluding tobacco-associated sites, the risk of a subsequent cancer was not significantly raised in either sex (SIR 0.87 (95% CI 0.81-0.93) for men; SIR 0.99 (95% CI 0.90-1.09) for women). CONCLUSIONS: The relative risk for multiple primary cancer was higher in younger subjects, those detected with a head and neck cancer during the 1990s as compared with earlier decades of the study, and among patients who received radiotherapy for their first tumour. By 20 years from the time of the first head and neck cancer, we estimate that approximately 30% of male patients and 20% of female patients will have developed an SPT. PMID- 12901723 TI - Biological effect of radiation-degraded alginate on flower plants in tissue culture. AB - Alginate with a weight-average molecular mass (Mw) of approx. 9.04 x 10(5) Da was irradiated at 10-200 kGy in 4% (w/v) aqueous solution. The degraded alginate product was used to study its effectiveness as a growth promoter for plants in tissue culture. Alginate irradiated at 75 kGy with an Mw of approx. 1.43 x 10(4) Da had the highest positive effect in the growth of flower plants, namely limonium, lisianthus and chrysanthemum. Treatment of plants with irradiated alginate at concentrations of 30-200 mg/l increased the shoot multiplication rate from 17.5 to 40.5% compared with control. In plantlet culture, 100 mg/l irradiated alginate supplementation enhanced shoot height (9.7-23.2%), root length (9.7-39.4%) and fresh biomass (8.1-19.4%) of chrysanthemum, lisianthus and limonium compared with that of the untreated control. The survival ratios of the transferred flower plantlets treated with irradiated alginate were almost the same as the control value under greenhouse conditions. However, better growth was attained for the treated plantlets. PMID- 12901725 TI - Loss of heterozygosity at APC and MCC genes of oral cancer and leukoplakia tissues from Indian tobacco chewers. AB - BACKGROUND: Loss of heterozygosity (LOH) at tumor suppressor genes, such as adenomatous polyposis coli (APC) and mutated in colon cancer (MCC) genes, is one of the early events in carcinogenesis of oral tissue in Caucasian and Chinese patients. We wanted to check whether it is also true in Indian oral pre-cancer and cancer patients. METHODS: Loss of heterozygosity at APC and MCC genes was investigated in 57 and 40 unrelated primary oral leukoplakia (a pre-cancerous lesion) and squamous cell carcinomas (SCC), respectively, by polymerase chain reaction. RESULTS: In these samples, most of the leukoplakia patients had tobacco smoking habit whereas majority of cancer patients had tobacco chewing habit. LOH at APC gene was observed in 4 of 16 (25%) and 1 of 29 (3%) informative tumor and leukoplakia DNAs from tobacco chewers, respectively. LOH at MCC gene was not detected either in tumor or in leukoplakia DNAs. CONCLUSION: This infrequent LOH at APC gene of pre-cancer and cancer tissues suggests that it may not be an early event in oral carcinogenesis in these patients. PMID- 12901726 TI - Expression of vascular endothelial growth factor-C correlates with the lymphatic microvessel density and the nodal status in oral squamous cell cancer. AB - BACKGROUND: The cause of preferential metastatic spreading to cervical lymph nodes in oral squamous cell cancer (SCC) is not quite clear. As the density of microvessels may influence the metastatic behaviour, we were interested in how the density of blood/lymphatic microvessels are related to primary SCC and the clinical course of the disease. METHODS: Lymphatic and blood microvessels of 28 patients with oral SCC were identified immunohistochemically by antibodies against podoplanin and CD34, respectively. Lymphatic microvessel density (LVD) and blood microvessel density (MVD), and the expression of VEGF-C were determined. These findings were compared with the long-term clinicopathological data of the patients. RESULTS: LVD and MVD were significantly higher than in control tissues. The amount of lymphatic microvessels correlated positively with the expression of VEGF-C, the tumour grade, the nodal status and with later appearing metastasis. The latter three parameters, however, did not influence the clinical course of the disease. CONCLUSIONS: VEGF-C expression in oral SCC triggers lymphatic angiogenesis, which may result in a higher risk for cervical lymph node metastasis. The angiogenetic effect of VEGF-C may also favour the onset of late lymphatic and haematogenous metastases. PMID- 12901727 TI - The biphasic differential expression of the cellular membrane protein, caveolin 1, in oral carcinogenesis. AB - BACKGROUND: The increased expression of Cav-1 is seen in various cancers from prostate, esophagus, colon, breast and pancreas yet the information regarding the correlation between the expression of Cav-1 and oral cancer is blind. Thus, the expression profile of caveolin-1 (Cav-1) in oral carcinogenesis and the correlation to the clinicopathologic covariates are examined in this study. METHODS: Immunohistochemistry was used to detect Cav-1 expression in non cancerous matched tissues (NCMT; n=12), and tissue from normal oral mucosa (NOM; n=12), oral pre-cancer lesions (OPL; n=17), primary oral squamous cell carcinoma (POSCC; n=47) and metastatic OSCC (MOSCC; n=8). The Cav-1 expression was correlated to the age, site, areca use, stage, size, nodal involvement, and differentiation stage. Western blot was used to confirm the specificity of antibody and to follow changes in Cav-1 expression. RESULTS: The Cav-1 immunoreactivity increased significantly from 8% in NOM and 17% in NCMT to 53% in OPL and 79% in POSCC. In addition, lymph node metastasis (LNM) was present in 62% of Cav-1(+) POSCCs, but only in 10% of Cav-1(-) POSCCs. Remarkably, only 38% of MOSCCs had Cav-1 immunoreactivity. CONCLUSION: An increased Cav-1 expression is seen in the stepwise carcinogenesis from NOM, NCMT, OPL to POSCC. The decrease in expression from the POSCC to MOSCC indicates the value to explore its biphasic functions in oral carcinogenesis. Whether Cav-1 is an important predictor or prognosis for survival still awaits the extension of clinical follow-up. PMID- 12901728 TI - Minichromosome maintenance 2 expression is correlated with mode of invasion and prognosis in oral squamous cell carcinomas. AB - BACKGROUND: This study examined the immunohistochemical expression of cell-cycle related molecules as well as cell proliferation and pathologic findings in oral squamous cell carcinoma (SCC) in order to clarify their pathobiologic and prognostic significance. METHODS: A total of 46 oral SCC specimens were analyzed using Ki-67, minichromosome maintenance 2 (MCM2), p53, p27, p21, and TUNEL. Aspects including tumor differentiation, mode of carcinoma invasion, tumor metastasis, and patient prognosis were compared among the specimens. RESULTS: A significantly higher MCM2 labeling index (LI) was observed in the moderately differentiated SCCs when compared to the well-differentiated SCCs (P<0.05). The higher MCM2 LI was correlated with mode of invasion Grade 4 (infiltrative growth) and patient prognosis. In contrast, the LIs of Ki-67, TUNEL-signal, p53, p27, and p21 were not correlated with patient prognosis. CONCLUSION: Higher MCM2 LI provides useful information for patient prognosis in oral SCCs. PMID- 12901729 TI - Innate immune system is implicated in recurrent aphthous ulcer pathogenesis. AB - BACKGROUND: Recurrent aphthous ulcers (RAU) is a chronic inflammatory disease with evidence of inappropriate immune response. This study presents the status of innate immune system in RAU. METHODS: Twenty RAU patients and 19 healthy individuals were selected. The status of peripheral blood neutrophils (reactive oxygen intermediates (ROI) production, CD11b, TNF-RI and TNF-RII expression), concentration of antioxidants, sTNF-R, C3c, C4 and haemolytic activity of the complement system, as well as mannose-binding lectin (MBL), in the serum of RAU patients in active stage and in remission of the disease were determined. RESULTS: Peripheral blood neutrophils were primed in RAU, which resulted in increased ROI production by resting and fMLP-stimulated neutrophils and diminished ROI production after in vitro priming. The increased expression of CD11b on resting and fMLP-stimulated neutrophils in RAU may also point to their previous in vivo stimulation. The decreased total antioxidant status of serum observed in RAU may be a result of increased ROI production by peripheral blood neutrophils. The levels of C3c, C4 and haemolytic activity of the complement system were higher in RAU than in healthy people. No significant differences between active and remission RAU were noted. CONCLUSION: Presented observations confirm that the innate immune system is involved in RAU pathogenesis. PMID- 12901730 TI - Oral nicorandil-induced lesions are not aphthous ulcers. AB - OBJECTIVES: (i) To accurately define these lesions determining whether oral nicorandil-induced lesions are aphthous ulcers; (ii) To determine clinical characteristics of oral nicorandil-induced lesions. MATERIALS AND METHODS: Two slide conferences were held. A total of 60 dermatologists assigned diagnosis and clinical criteria to 11 photographs of oral nicorandil-induced lesions. Two slides were randomly selected and duplicated to be used as control. The panel of slides included independent lesions and photographs of different lesions of the same patient. Statistical analysis used chi2-test, estimation of the percentage interobserver agreement, and kappa-values. RESULTS: The diagnosis of non-aphthous ulcer (71.8%) was significantly held in comparison with the diagnosis of oral aphthous ulcer (28.2%; P<0.001). To differentiate aphthous ulcer from non aphthous ulcer, physicians significantly used three clinical criteria. The diagnosis of non-aphthous ulcer were significantly associated with the absence of yellow-based ulceration (P<0.001), with the linear shape (P=0.006) and the absence of inflammatory halo (P=0.003). CONCLUSION: Oral nicorandil-induced lesions are not aphthous ulcers. We propose that at this stage of our knowledge, oral nicorandil-induced ulcer is the most suitable terminology. PMID- 12901731 TI - Dentigerous cyst versus unicystic ameloblastoma--differential diagnosis in routine histology. AB - BACKGROUND: Unicystic ameloblastomas (UAs) and dentigerous cysts (DCs) have an identical clinical and radiographic appearance. Some subtypes of UAs have a better prognosis than solid or multicystic ameloblastomas, and simple enucleation is the adequate treatment. The present study was designed to test the hypothesis that UAs with small islands of ameloblastomatous epithelium may be misdiagnosed as a DC or keratocyst if no more than two histologic sections are examined. METHODS: A total of 101 resection specimens from 22 women and 73 men (mean age: 46.5 years) were selected, all showing the clinical and radiographic features of a DC. Only cysts with a minimum diameter of 15 mm in the panoramic X-ray were considered for the present investigation. The histopathologic diagnosis had been routinely established by examining two sections. For our study, the specimens were investigated by step sections at 50 microm and by staining of 5 microm thin sections with hematoxylin and eosin (H&E) at 1 mm levels. An average of 15 slides were evaluated per case. RESULTS: Microscopic examination of the step sections did not reveal ameloblastomatous epithelium in the cyst lining epithelium of the 101 cases. Thus, every primary diagnosis of a dentigerous cyst was confirmed. In four cases, additional rather large odentogenic cell nests were detected with palisading of basaloid cells, while there was a lack of other signs of ameloblastic differentiation. All lesions were completely resected, and no additional treatment was performed. CONCLUSIONS: Step sectioning of larger DCs may reveal associated odontogenic cell nests in some cases but does not lead to the detection of formerly missed ameloblastic cells. Thus, unicystic ameloblastomas are not misdiagnosed if only two slides are prepared for routine diagnosis of DCs. PMID- 12901732 TI - Endovascular papillary angioendothelioma (Dabska tumor) of the tongue: report of a case. AB - Endovascular papillary angioendothelioma (EPA), known as Dabska tumor, is a very rare vascular neoplasm that usually occurs in the skin or subcutis of infants and young children. There is no previous report of EPA appearing in the oral cavity. Here, we present an exceptionally rare case of EPA of the tongue in a 67-year-old man. A well-defined, reddish tumor measuring 11 mm x 8 mm x 7 mm existed at the submucosal area of the left tongue margin. Microscopic examination of an excisional biopsy specimen revealed the endothelioid tumor cells showing a papillary growth pattern, such as blood vessel-like structures. Immunohistochemical studies showed positive reactivities for CD31, CD34, alpha smooth muscle actin, and factor VIII-related antigen in most of the tumor cells and CD68 in some tumor cells. Based on these observations, the tumor was diagnosed as an EPA. PMID- 12901733 TI - Maxillary odontogenic keratocyst with respiratory epithelium: a case report. AB - We report a case of odontogenic keratocyst with a respiratory epithelial lining and a malformed impacted tooth in the maxilla of a 39-year-old Japanese female who suffered from swelling symptoms for half a year. CT examinations revealed an air-filled cystic lesion with an impacted tooth crown in the maxillary bone which expanded to the nasal cavity as well as to the maxillary sinus. Histopathologically, the surgically removed cyst wall consisted of fibrous granulation tissue with a lining of parakeratinized squamous epithelium as well as ciliated pseudostratified epithelium and with retention of desquamated keratin materials in the lumen. The impacted tooth was malformed lacking a root portion. We discuss the frequency of respiratory epithelium in odontogenic keratocysts. PMID- 12901734 TI - Pleomorphic adenoma of the submandibular salivary glands with marked ossification. AB - This article describes an unusual case of pleomorphic adenoma arising in the submandibular salivary glands that was associated with marked ossification. CT scan showed an irregular high-density mass in the left submandibular region. Histologic examination of the resected mass revealed a well-circumscribed pleomorphic adenoma with chondroid tissue intermingled with glandular and trabecular epithelial components. Irregularly formed osseous tissue was present in the chondroid tissue. The osseous tissue showed bone matrix on von Kossa's staining. Alkaline phosphatase expression was restricted to the vicinity of osseous tissue. Tartrate-resistant acid phosphatase-positive osteoclasts were observed at the periphery of the osseous tissue. CD34 staining demonstrated many microvascular vessels in both the chondroid and osseous tissues. We conclude the osseous tissue in pleomorphic adenoma was bone in nature, and via the process of endochondral ossification most likely depended on angiogenesis. PMID- 12901735 TI - Intra-oral Dirofilaria repens infection: report of seven cases. AB - Cutaneous dirofilariasis usually affects animals such as cats and dogs which are known to be the natural host of Dirofilaria. Dirofilariasis displays a worldwide distribution. Certain geographic regions account for the majority of reported cases. South-eastern United States, Australia and Europe have been identified as endemic regions (1, 2). However, new endemic areas are arising with increased awareness in African and Asian regions. Out of about 40 different species of Dirofilaria only a few species are commonly known to infect man, namely Dirofilaria immitus, D. tenices and D. repens. Human infection occurs when a human becomes a blood meal of an infected arthropod vector and on accidental entering of the worm. Rarely this zoonotic infection affects the oral mucosa (3). Man is the dead end of this parasite. Since the human body is an abnormal environment for the parasite, the development of the larvae is inhibited by means of retardation of sexual maturity. Lesions are presented as single non-tender subcutaneous nodules, and most patients are asymptomatic. Diagnosis is established by H&E sections prepared from excised nodules. In a majority of the cases, only a single worm either a male or a female could be identified. The worm is usually dead and degenerated with a massive inflammatory cell infiltration. Seven new cases presented as intra-oral nodules with their clinicopathological correlation are discussed. PMID- 12901736 TI - Distribution of the two forms of Plasmodium falciparum erythrocyte binding antigen-175 (eba-175) gene in Lao PDR. AB - BACKGROUND: The erythrocyte binding antigen 175 (EBA-175) is a 175 kDa antigen of Plasmodium falciparum and plays a major role in erythrocyte recognition by the parasite. The antigen is also supposed to be partly responsible for the invasion of erythrocytes by merozoites. EBA-175 has been sequenced from the FCR-3 and CAMP strains of P. falciparum. The sequences were identical in most parts of the gene. Differences were apparent in a 423 bp segment in the FCR-3 strain, the F Fragment, that is not found in the CAMP-strain and a 342 bp segment, the C Fragment, which is present in the CAMP-strain but not in the FCR-3-strain. The aim of this study was to assess the distribution of the two EBA-175-alleles in the Lao PDR. MATERIALS & METHODS: Altogether, 240 blood-samples were collected in two areas of the country: Attapeu in the south and Lung Namtha in the north. Subsequently, the material was scanned for the F-and C-fragments. RESULTS: In the whole study population, 52% carried the F-fragment, and 41% the C-fragment while seven percent of the patients were infected with at least two parasite strains and showed both alleles. CONCLUSION: Distribution of the alleles showed significant differences between the north and the south province. Reasons for this include possible importation of different parasite strains from neighbouring countries. PMID- 12901737 TI - The new variant of the Creutzfeldt-Jakob disease accounts for no relative increase of the Creutzfeldt-Jakob disease mortality rate in the United Kingdom; this fits ill with the new variant being the consequence of consumption of food infected with the agent of Bovine Spongiform Encephalopathy. AB - BACKGROUND: A new variant of Creutzfeldt-Jakob disease was described in the United Kingdom. It is often claimed that it is caused by consumption of food infected with the agent of bovine spongiform encephalopathy. However, this remains open to question because the number of cases of the variant is, at the present time, less than would be expected from a major food-borne source. DISCUSSION: The EUROCJD cooperative study presents currently available epidemiological data of Creutzfeldt-Jakob disease and its new variant, for nine European countries plus Australia and Canada. Unexpectedly, for the United Kingdom where all but a few cases of the new variant have been reported, these cases have to be included in the incidence curve of the sporadic forms of the disease in order to obtain the best fit with the median curve from all the countries. This variant could be merely a rare clinical phenotype within the sporadic disease. The published clinical and experimental data which suggest that it is linked with bovine spongiform encephalopathy, lead us to propose that this link could be a common etiological origin other than consumption of bovine infected food. In any case, public health recommendations hold and further investigation is required. SUMMARY: The lack of a relative increase of the Creutzfeldt-Jakob-disease mortality rate in the United Kingdom, does not fit well with the new variant being the consequence of consumption of food infected with the agent of bovine spongiform encephalopthy. PMID- 12901738 TI - Sinus versus nonsinus tachycardia in the emergency department: importance of age and heart rate. AB - BACKGROUND: The emergency department diagnosis of sinus versus nonsinus tachycardia is an important clinical challenge. The objective of this study was to identify subjects with a high prevalence of nonsinus tachycardia. METHODS: Heart rate and cardiac rhythm were prospective reviewed in 500 consecutive patients with heart rate > or = 100 beats/min in a busy emergency department. A predictive model based on age and heart rate was then developed to identify the probability of nonsinus tachycardia. RESULTS: As age and heart rate increased, nonsinus tachycardias became more frequent. The probability of nonsinus tachycardia in a subject > or = 71 years with heart rate > or = 141 beats/minute was 93%, compared to only three percent in a subject < or = 50 years with heart rate 100-120 beats/minute. A simple point score system based on age and heart rate helps predict the probability of sinus tachycardia versus nonsinus tachycardia. CONCLUSION: Nonsinus tachycardia is significantly more common than sinus tachycardia in elderly patients in the emergency department. The diagnosis of sinus tachycardia becomes much less likely as age and heart rate increase. PMID- 12901759 TI - Key events in the history of electron microscopy. PMID- 12901761 TI - High resolution CryoFESEM of microbial surfaces. AB - The outer surfaces of three microorganisms, Giardia lamblia, Enterococcus faecalis, and Proteus mirabilis, were investigated by cryo-immobilization followed by sublimation of extracellular ice and cryocoating with either Pt alone or Pt plus carbon. Cryocoated samples were examined at -125 degrees C in either an in-lens field emission SEM or a below-the-lens field emission SEM. Cryocoating with Pt alone was sufficient for low magnification observation, but attempts to do high-resolution imaging resulted in radiolysis and cracking of the specimen surface. Double coating with Pt and carbon, in combination with high resolution backscatter electron detectors, enabled high-resolution imaging of the glycocalyx of bacteria, revealing a sponge-like network over the surface. High resolution examination of bacterial flagella also revealed a periodic substructure. Common artifacts included radiolysis leading to "cracking" of the surface, and insufficient deposition of Pt resulting in the absence of detectable surface topography. PMID- 12901762 TI - Cryo-fracturing and cryo-planing for in-lens cryo-SEM, using a newly designed diamond knife. AB - High-pressure frozen rat pancreas tissue samples were cryo-fractured and cryo sectioned with a diamond knife in the microtome of a freeze-etching device. The bulk fracture faces and block faces were investigated in the frozen-hydrated state by use of a cryo-stage in an in-lens SEM. With this combination, relevant biological structures can be investigated with a few nanometers resolution in a near lifelike state, preventing the artifacts of conventional fixation techniques. Compared to TEM replica techniques, the presented method is more versatile since no replica cleaning is necessary. Additional structures can be made visible by controlled sublimation of ice, leading to a better understanding of the three-dimensional organization of organelles, such as the endoplasmic reticulum. PMID- 12901763 TI - In-lens cryo-high resolution scanning electron microscopy: methodologies for molecular imaging of self-assembled organic hydrogels. AB - The micro- and nanoarchitectures of water-swollen hydrogels were routinely analyzed in three dimensions at very high resolution by two cryopreparation methods that provide stable low-temperature specimens for in-lens high magnification recordings. Gemini surfactants (gS), poly-N-isopropylacrylamides (p NIP Am), and elastin-mimetic di- (db-E) and triblock (tb-E) copolymer proteins that form hydrogels have been routinely analyzed to the sub-10-nm level in a single day. After they were quench or high pressure frozen, samples in bulk planchets were subsequently chromium coated and observed at low temperature in an in-lens field emission SEM. Pre-equilibrated planchets (4-40 degrees C) that hold 5-10 microl of hydrogel facilitate dynamic morphological studies above and below their transition temperatures. Rapidly frozen samples were fractured under liquid nitrogen, low-temperature metal coated, and observed in-lens to assess the dispersion characteristics of micelles and fragile colloidal assemblies within bulk frozen water. Utilizing the same planchet freezing system, the cryoetch HRSEM technique removed bulk frozen water from the hydrogel matrix by low temperature, high-vacuum sublimation. The remaining frozen solid-state sample faithfully represented the hydrogel matrix. Cryo- and cryoetch-HRSEM provided vast vistas of hydrogels at low and intermediate magnifications whereas high magnification recordings and anaglyphs (stereo images) provided a three dimensional prospective and measurements on a molecular level. PMID- 12901764 TI - Automated three-dimensional tracing of neurons in confocal and brightfield images. AB - Automated three-dimensional (3-D) image analysis methods are presented for tracing of dye-injected neurons imaged by fluorescence confocal microscopy and HRP-stained neurons imaged by transmitted-light brightfield microscopy. An improved algorithm for adaptive 3-D skeletonization of noisy images enables the tracing. This algorithm operates by performing connectivity testing over large N x N x N voxel neighborhoods exploiting the sparseness of the structures of interest, robust surface detection that improves upon classical vacant neighbor schemes, improved handling of process ends or tips based on shape collapse prevention, and thickness-adaptive thinning. The confocal image stacks were skeletonized directly. The brightfield stacks required 3-D deconvolution. The results of skeletonization were analyzed to extract a graph representation. Topological and metric analyses can be carried out using this representation. A semiautomatic method was developed for reconnection of dendritic fragments that are disconnected due to insufficient dye penetration, an imaging deficiency, or skeletonization errors. PMID- 12901765 TI - Microscopic aspects of autoschizic cell death in human ovarian carcinoma (2774) cells following vitamin C, vitamin K3 or vitamin C:K3 treatment. AB - Human ovarian carcinoma cells (MDAH 2774) were treated with sodium ascorbate (VC), menadione (VK3), or with a VC:VK3 combination for 1 h and then studied using light microscopy (LM) and scanning (SEM) and transmission electron (TEM) microscopy. Plasma membrane damage (blisters and blebs, hairy aspect) results from vitamin C (VC) treatment, while cytoskeletal damage and self-morsellation are caused by vitamin K3 (VK3) treatment. VC:VK3-treated cells exhibit exacerbated injuries characteristic of both VC and VK3 treatment as well as a significant decrease in cell diameters from 20-35 microm for control cells to 7 12 microm for VC:VK3 treatment. Moreover, after a 1-h exposure to the vitamin combination, autoschizis (43%), apoptosis (3%), and oncosis (1.9%) are observed at the percentages indicated. All cellular changes associated with autoschizis observed with SEM were confirmed by LM and TEM observations and are consistent with cell death by autoschizis: decrease in cell size, cytoplasmic self excisions, degradation of the nucleus and nucleolus without formation of apoptotic bodies and, ultimately, karyorrhexis and karyolysis. These results also suggest that the vitamin combination may find clinical use in the treatment of ovarian cancer. PMID- 12901768 TI - Characterization of tungsten surfaces by simultaneous work function and secondary electron emission measurements. AB - The changes of the work function (Phi) and the secondary electron emission (SEE) of oxygen covered polycrystalline tungsten occurring after ion sputtering and heat treatments have been investigated. The chemical composition was analyzed by X-ray photoelectron spectroscopy (XPS), and the electron emission properties by work function spectroscopy (WFS). We observed in what manner the chemical changes of the surface are reflected in the work function and SEE. The simultaneous change of Phi and SEE in the case of oxygen covered tungsten have been pointed out and a direct relationship between them can be supposed. PMID- 12901766 TI - Structural evidence for actin-like filaments in Toxoplasma gondii using high resolution low-voltage field emission scanning electron microscopy. AB - The protozoan parasite Toxoplasma gondii is representative of a large group of parasites within the phylum Apicomplexa, which share a highly unusual motility system that is crucial for locomotion and active host cell invasion. Despite the importance of motility in the pathology of these unicellular organisms, the motor mechanisms for locomotion remain uncertain, largely because only limited data exist about composition and organization of the cytoskeleton. By using cytoskeleton stabilizing protocols on membrane-extracted parasites and novel imaging with high-resolution low-voltage field emission scanning electron microscopy (LVFESEM), we were able to visualize for the first time a network of actin-sized filaments just below the cell membrane. A complex cytoskeletal network remained after removing the actin-sized fibers with cytochalasin D, revealing longitudinally arranged, subpellicular microtubules and intermediate sized fibers of 10 nm, which, in stereo images, are seen both above and below the microtubules. These approaches open new possibilities to characterize more fully the largely unexplored and unconventional cytoskeletal motility complex in apicomplexan parasites. PMID- 12901769 TI - Calculation of the surface excitation parameter for Si and Ge from measured electron backscattered spectra by means of a Monte-Carlo simulation. AB - A new Monte Carlo method has been developed for simulating backscattered electron spectra, and this was applied for determining the surface excitation parameter (SEP). The simulation is based on direct tracking of electron trajectories in the solid, taking into account elastic and inelastic events. The elastic scattering cross sections are taken from literature, while inelastic cross section data are obtained by a fitting procedure. After some iterations, the program produces electron spectra fitting well to the experimental ones. Si and Ge electron spectra were simulated and SEP values were calculated. The SEP values are compared to other ones from literature. PMID- 12901770 TI - Investigation of winter atmospheric aerosol particles in downtown Katowice using XPS and SEM. AB - X-ray photoelectron spectroscopy (XPS) was used to determine the surface chemical composition of atmospheric particles (PM-10 and PM-2.5), collected in downtown Katowice, Upper Silesia, Poland, in the winter season (November and December 2000). Carbon- and oxygen-containing species dominated the particulate surface with traces of N, S, Si, Cl, Na, Zn, Al, Cu, Fe, Ca, K, Mg, Pb, and P present. Additionally, the size, morphology, and chemical composition of about 300 of the individual atmospheric particles were analyzed by high-resolution scanning electron microscopy and electron probe microanalysis. A number of aluminosilicates and metallic elements such as Fe, K, Mg, Zn, as well as rare earth elements, were detected by SEM. The results obtained show that the analyzed aerosol is of natural and anthropogenic origin. Particles containing sulfur compounds as well as oxygen and sodium in downtown Katowice come mainly from the east and southeast sectors. PMID- 12901771 TI - Electron probe and auger electron microprobe characterization of modified Cu based amorphous alloys. AB - Changes in morphology and local chemical composition due to various methods of modification of surfaces of Cu-Zr, Cu-Hf, and Cu-Ti amorphous alloys (caused by aging in air/dry corrosion or hydrogen charging) were investigated. These modification/activation procedures transform the original amorphous ribbons of low surface area into efficient and stable catalysts, due to the segregation of a distinct amount of Cu and the development of a large specific surface area of Cu on a ZrO x or HfO x support. It was found that aging in air resulted in the formation of a bilayer of rough copper (containing small Cu particles indispensable for catalysis) on top of a rather smooth oxide underlayer (ZrO x, HfO x ). Careful examination of the cross sections of the modified Cu-based ribbons revealed that, even after prolonged aging in air, only the first few microns of the surface layer was modified. Cu-Ti alloy was stable in air and did not undergo the expected modification. Hydrogenation followed by air exposure resulted in a disintegration of the ribbons into small pieces. Each piece was covered with many small Cu clusters 0.1-0.5 microm in diameter formed on an oxide underlayer. High-energy resolution Auger spectroscopy allowed identification of the underlayers (ZrO2, HfO2, or TiO x ), identification of small Cu clusters, determination of the degree of surface oxidation of them, and mapping of the surface to identify the Cu-covered and "naked" heavy metal. PMID- 12901774 TI - Non-acute (residual) neurocognitive effects of cannabis use: a meta-analytic study. AB - The possible medicinal use of cannabinoids for chronic diseases emphasizes the need to understand the long-term effects of these compounds on the central nervous system. We provide a quantitative synthesis of empirical research pertaining to the non-acute (residual) effects of cannabis on the neurocognitive performance of adult human subjects. Out of 1,014 studies retrieved using a thorough search strategy, only 11 studies met essential a priori inclusion criteria, providing data for a total of 623 cannabis users and 409 non- or minimal users. Neuropsychological results were grouped into 8 ability domains, and effect sizes were calculated by domain for each study individually, and combined for the full set of studies. Using slightly liberalized criteria, an additional four studies were included in a second analysis, bringing the total number of subjects to 1,188 (i.e., 704 cannabis users and 484 non-users). With the exception of both the learning and forgetting domains, effect size confidence intervals for the remaining 6 domains included zero, suggesting a lack of effect. Few studies on the non-acute neurocognitive effects of cannabis meet current research standards; nevertheless, our results indicate that there might be decrements in the ability to learn and remember new information in chronic users, whereas other cognitive abilities are unaffected. However, from a neurocognitive standpoint, the small magnitude of these effect sizes suggests that if cannabis compounds are found to have therapeutic value, they may have an acceptable margin of safety under the more limited conditions of exposure that would likely obtain in a medical setting. PMID- 12901775 TI - Effects of the use of alcohol and cigarettes on cognition in elderly African American adults. AB - In this study we examined the independent and interactive effects of lifetime patterns of drinking and smoking on cognitive performance in elderly African Americans. A sample of 230 individuals with varying histories of alcohol and cigarette use was drawn from the Hillsborough Elder African American Life Study, a community-based, cross-sectional study of older adults aged 60 to 84. Dependent variables were the results of a neuropsychological battery that provided measures of general cognitive ability, executive function, and memory. Specifically, our study addressed (1) whether individuals with a lifetime history of sustained smoking and/or drinking show lower levels of cognitive performance in comparison to lifetime abstainers, (2) whether cumulative lifetime doses of alcohol or cigarettes, or of the two substances in interaction, have an effect on cognition, and (3) whether individuals who have histories of periodic, intense use of either alcohol or cigarettes show lower levels of cognitive performance in comparison to lifetime abstainers. When significant results were obtained, effect sizes were small, not exceeding 5% of the variance. A single exception occurred for the intensity analyses, in which drinking explained approximately 16% of the variance in global cognitive ability after adjusting for the contributions of control variables. In these analyses, drinking was found to have a U-shaped effect on global cognitive ability and total acquisition in the memory trials. Specifically, moderate users performed at a lower level than abstainers or heavy users, who did not differ from each other. PMID- 12901776 TI - Relationship between positive and negative symptoms and neuropsychological scores in frontotemporal dementia and Alzheimer's disease. AB - Patients with dementia, particularly those with frontotemporal dementia (FTD), are reported to display marked negative symptoms, including apathy, lack of initiative, and flattened affect, similar to those observed in schizophrenic patients. However, negative symptoms have yet to be formally quantified in an FTD population. Twenty-seven patients with FTD (11 primarily right-sided, 8 primarily left-sided, and 4 symmetric) and 7 patients with Alzheimer's disease were rated on the Scale for the Assessment of Negative Symptoms, the Positive and Negative Syndrome Scale, and the Emotional Blunting scale. The FTD patients registered significantly more negative symptoms than the Alzheimer's patients, averaging a threefold increase; groups did not significantly differ in positive symptoms. Negative symptom scale scores were negatively correlated with nonverbal executive skills (23-44% shared variance), verbal executive skills (up to 25% shared variance) and verbal memory (up to 20% shared variance), but were unrelated to measures of attention, verbal and nonverbal information processing, nonverbal memory, language, and constructional skill. In contrast, positive symptoms were positively correlated with constructional skill (19% shared variance) and attentional scores (15% shared variance). These findings add to the existing literature relating negative symptoms to anterior cerebral hypofunction, and suggest that positive symptoms, at least in this population, may be tied to increased posterior activation. PMID- 12901777 TI - Primary progressive aphasia: diagnosis, varieties, evolution. AB - A referred cohort of 67 clinically defined PPA patients were compared to 99 AD patients with formal language and nonverbal cognitive tests in a case control design. Language fluency was determined at the first and last follow up visits. Quantitation of sulcal and ventricular atrophy on MRI was carried out in 46 PPA and 53 AD patients. Most PPA patients (57%) are relatively fluent when first examined. Visuospatial and memory functions are initially preserved. Aphemic, stuttering, "pure motor" presentation, or agrammatic aphasia are seen less frequently. Later most PPAs become logopenic and nonfluent, even those with semantic aphasia (dementia). In contrast, AD patients were more fluent and had relatively lower comprehension, but better overall language performance. MRI showed significant left sided atrophy in most PPA patients. Subsequent to PPA, 25 patients developed behavioral manifestations of frontotemporal dementia and 15 the corticobasal degeneration syndrome, indicating the substantial clinical overlap of these conditions. Language testing, particularly fluency scores supported by neuroimaging are helpful differentiating PPA from AD. The fluent nonfluent dichotomy in PPA is mostly stage related. The aphemic-logopenic agrammatic and semantic distinction is useful, but the outcomes converge. PMID- 12901778 TI - The neuropsychological profiles of mild Alzheimer's disease and questionable dementia as compared to age-related cognitive decline. AB - Test scores from a comprehensive neuropsychological battery administered to 1602 subjects consisting of 1347 subjects with probable Alzheimer's disease (AD), 100 subjects with questionable dementia (QD) and 155 non-demented elderly control subjects were cross-sectionally analyzed. Subjects with probable AD were categorized as mild (n = 244), moderate (n = 480), severe (n = 376), and very severe (n = 247) according to modified mini mental status exam (mMMSE) scores. Mean scores on individual neuropsychological tests are provided for each group of subjects. Stratified random sampling was performed to select a sample of mild AD subjects who were matched in age and education to non-demented elderly controls, and analyses focused on the performance of QD subjects and mild AD subjects, whose scores were compared to those of the elderly control subjects. Selected scores were organized by cognitive domain and logistic regressions were used to determine the domains and individual tests within each that were most predictive of group status. Results suggested a profile of scores associated with QD and mild AD including impaired recall of verbal information for both groups. Areas of lower functioning in QD subjects as compared to elderly controls included category fluency and visuospatial ability. PMID- 12901779 TI - Narrative and procedural discourse in temporal lobe epilepsy. AB - It is well established that some individuals with temporal lobe epilepsy (TLE) demonstrate language deficits at the single word level. However, discourse production rarely has been examined quantitatively within this group. This study compared adult TLE patients with an early seizure onset (< or = age 14 years, n = 27) to a control group (n = 28) on narrative and procedural discourse tasks. As a group, the TLE patients performed normally on the procedural discourse task, but differed significantly from the controls on several narrative discourse variables. At the individual level, 30% of the TLE patients versus 4% of the controls demonstrated impaired discourse ability (p and 0.01). Within this early onset TLE group, discourse performance was not associated with demographic or seizure history variables. Considering the cognitive domain, discourse performance correlated significantly with working memory. In summary, mild discourse dysfunction was present in a significant minority of early onset TLE patients, but this deficit was not closely associated with other language measures. Discourse ability and its neuropsychological, neuroanatomical and conversational speech correlates deserve further study in TLE patients. PMID- 12901780 TI - Severe hypoglycemia and long-term spatial memory in children with type 1 diabetes mellitus: a retrospective study. AB - Performance on long delays of delayed response tasks is associated with medial temporal function, a region of the brain affected by severe hypoglycemia. A previous study showed that children with type 1 diabetes mellitus (T1DM) with higher risk for severe hypoglycemia performed worse than controls on long delays of a spatial delayed response (SDR) task. We tested the more specific hypothesis that frequency of severe hypoglycemia would relate to long delay SDR performance. Children with T1DM (n = 51) and controls (n = 32) performed the SDR task with short and long delays. Information was collected on children's past severe hypoglycemia. In children with T1DM, number of past severe hypoglycemic episodes accounted for a significant portion of the variance in long delay SDR after controlling for age and age of onset. This relationship was not seen with short delay SDR or with other tasks (verbal or object memory, attention, motor speed). These results support the hypothesis that severe hypoglycemia has specific, negative effects on memory skills in children. If this relationship is extrapolated to children with higher frequency of severe hypoglycemia, due to longer duration of disease or poorer glucose control, it may affect daily functioning and thus need to be considered in treatment decisions. PMID- 12901781 TI - Cognitive asymmetries associated with apolipoprotein E genotype in patients with Alzheimer's disease. AB - The relationship between apolipoprotein E (apoE) genotype and cognitive performance was examined in 200 patients with probable Alzheimer's disease (AD). Differences between composite measures of verbal and nonverbal functioning were used to define asymmetric patterns of cognition. Patients who were homozygous for apoE epsilon4 demonstrated relatively worse nonverbal as compared to verbal cognitive ability. In contrast, participants who were heterozygous for apoE epsilon4 or who possessed no epsilon4 allele demonstrated relatively equivalent verbal and nonverbal cognitive abilities. Although age and dementia severity also contributed to these patterns, apoE genotype appears to have a significant unique contribution to cognitive performance in these individuals. The epsilon4 allele may thus be associated with a specific neurocognitive phenotype among patients with AD, with the overall pattern of cognitive asymmetry dependent upon epsilon4 dose. PMID- 12901782 TI - Impulsivity and verbal deficits associated with domestic violence. AB - While neurobiological factors are known to play a role in human aggression, relatively few studies have examined neuropsychological contributions to propensity for violence. We previously demonstrated cognitive deficits among men who committed domestic violence (batterers) compared to non-violent controls. Batterers had deficits in verbal ability, learning and executive problem-solving ability. These findings led us to examine whether executive control problems involving impulsivity contribute to problems with behavioral control among batterers, and to further examine their deficits in verbal functioning. Batterers (n = 41) enrolled in a domestic violence program were compared to 20 non-violent men of similar age, education, and socioeconomic background on neuropsychological tests of executive functioning, including impulsivity. Questionnaires and structured clinical interviews were used to assess emotional distress, aggression and self-reported impulsivity. Batterers showed greater impulsivity compared to non-batterers on several neuropsychological measures. Yet, the severity of these deficits was relatively mild and not evident in all batterers. Consistent with our previous findings, significant verbal deficits were again observed among the batterers. These findings suggest that while impulsivity may be a factor associated with domestic violence, it probably is not the sole determinant of the strong relationship between cognitive functioning and batterer status that we previously observed. Both verbal expressive deficits and behavioral impulsivity appear to be relevant variables in predisposing men to domestic violence. PMID- 12901783 TI - Cognitive consequences of early phase of literacy. AB - The effect of the degree of illiteracy (complete or incomplete) on phonological skills, verbal and visual memory and visuospatial skills is examined in 97 normal Brazilian adults who considered themselves illiterate, and 41 Brazilian school children aged 7 to 8 years, either nonreaders or beginning readers. Similar literacy effects were observed in children and in adults. Tasks involving phonological awareness and visual recognition memory of nonsense figures distinguish the best nonreaders and beginning readers. Children performed better than adults at oral repetition of short items and figure recall, and adults better than children at semantic verbal fluency, digit span, and word list recall. A principal component analysis of the correlations between tasks showed that phonological awareness/reading, phonological memory/oral repetition, and semantic verbal memory/fluency tasks, generated different components. The respective role of culturally based preschool activities and literacy on the cognitive functions that are explored in this study is discussed. PMID- 12901785 TI - Semantic category differences in cross-form priming. AB - Findings of category-specific impairments have suggested that human semantic memory may be organized around a living/nonliving dichotomy. In order to assess implicit memory performance for living and nonliving concepts, one group of neurologically intact individuals participated in a cross-form conceptual priming paradigm. In Block 1, pictures primed words while in Block 2 words were used to prime pictures. Across all phases of the experiment, subjects decided whether items represented something which was living or nonliving, and response times were recorded. Results revealed greater priming for living concepts across both blocks. Greater priming for living concepts may have occurred because of increased or prolonged conceptual activation of these concepts. Results are discussed in the context of theoretical accounts of the category-specific impairments observed in brain-damaged populations. PMID- 12901784 TI - Neuropsychological deficits associated with Alzheimer's disease in the very-old: discrepancies in raw vs. standardized scores. AB - The profiles of neuropsychological deficits associated with Alzheimer's disease (AD) in Young-Old (M age and 70) and Very-Old (M age > 80) patients were compared, along with possible modifying effects of apolipoprotein E (APOE) genotype on these profiles. A comprehensive battery of neuropsychological tests was administered to the two AD patient groups (Young-Old: n = 33; Very-Old: n = 48) and their respective age-matched normal control (NC) groups who remained free of dementia on follow-up examinations over a 1 to 10 year period (Young-Old: n = 43; Very-Old: n = 36). AD and NC groups did not differ in education levels or gender distributions. Young-Old AD and Very-Old AD groups were comparable in education, gender, dementia severity, and disease duration. Results showed that both AD groups achieved comparable raw scores on all the neuropsychological measures. However, when scores were standardized on the basis of performance of their respective NC groups (i.e., age-corrected z scores), Very-Old AD patients significantly outperformed Young-Old AD patients on tests of executive functions, visuospatial skills, and delayed memory. Furthermore, the relationship between age and memory and executive function deficits in AD was modified by APOE genotype. These data suggest that the profile of neuropsychological deficits associated with AD in the Very-Old lacks the disproportionate saliency of episodic memory and executive function deficits typical of the Young-Old. PMID- 12901786 TI - Moyamoya disease in a patient with schizophrenia. AB - We present the case of a 23-year-old Vietnamese male with a 2-year history of a psychotic illness marked by prominent negative symptoms, fatuousness and disturbed behavior. Neuroimaging revealed a prominent vascular flow void affecting the middle and anterior cerebral arteries, with associated increased collateral supply to the frontal cortex, consistent with Moyamoya disease. Neurological examination was unremarkable; however, neuropsychological assessment revealed significant executive dysfunction, including stimulus-driven behavior. Whilst the diagnosis of schizophrenia and Moyamoya disease may be coincidental, an interaction between the 2 diseases may have led to some of the atypical features of this case, including prominent executive dysfunction and marked sensitivity to psychotropic medication. We discuss the nature of possible interactions between the 2 conditions. This case also highlights the importance of re-evaluating patients with atypical or treatment-resistant psychoses for cerebral pathology. PMID- 12901787 TI - Are perceptual and motor inhibition processes really dissociated? A comment on Nassauer and Halperin (2003). PMID- 12901789 TI - Nectins and nectin-like molecules: roles in cell adhesion, migration, and polarization. AB - Nectins are a family of Ca(2+)-independent immunoglobulin-like cell-cell adhesion molecules consisting of four members, which homophilically and heterophilically trans-interact and cause cell-cell adhesion. Nectin-based cell-cell adhesion is involved in the formation of cadherin-based adherens junctions in epithelial cells and fibroblasts. The nectin-based cell-cell adhesion induces activation of Cdc42 and Rac small G proteins, which eventually regulate the formation of adherens junctions through reorganization of the actin cytoskeleton, gene expression through activation of a mitogen-activated protein kinase cascade, and cell polarization through cell polarity proteins. Five nectin-like molecules (necls), which have domain structures similar to those of nectins, have recently been identified and appear to play different roles from those of nectins. One of them, named necl-5, which does not homophilically trans-interact, but heterophilically trans-interacts with nectin-3, regulates cell migration and adhesion. In this article, the roles and modes of action of nectins and necls in cell adhesion, migration, and polarization are reviewed. PMID- 12901790 TI - The D4Mit12 locus on mouse chromosome 4 provides susceptibility to both gamma-ray induced and N-methyl-N-nitrosourea-induced thymic lymphomas. AB - Low-penetrance genes control different susceptibilities to gamma-ray-induced thymic lymphomas in mouse strains. Our previous genetic analyses with backcross mice between BALB/c and MSM strains and congenic lines localized one such gene near the D4Mit12 locus on chromosome 4. N-Methyl-N-nitrosourea (MNU) is a guanine base-alkylating agent and differs from gamma-radiation in its mechanism of mutagenic action. Accordingly, in this study, we examined whether or not the locus also provides susceptibility to MNU-induced thymic lymphomas using 84 offsprings derived from congenic mice for D4Mit12. Association analysis provided a suggestive linkage at D4Mit12 (P = 0.0075) and the linkage was sustained by the peak of likelihood ratio statistical values being at the same position as that for the gamma-ray-induced lymphomas. The results strongly suggest that the BALB/c allele near D4Mit12 is associated with susceptibility to lymphomas induced by two carcinogenic agents having different mechanisms of mutagenic action. PMID- 12901791 TI - Differential gene expression profiles in colon epithelium of two rat strains with distinct susceptibility to colon carcinogenesis after exposure to PhIP in combination with dietary high fat. AB - Colon cancers develop through accumulation of multiple genetic and epigenetic alterations in colon epithelial cells, and the environment of the genetically altered epithelial cells may also have a substantial impact on their further development to cancer. In the present study, groups of 6-week-old F344 and ACI male rats, the former strain being susceptible to colon carcinogenesis induced by 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) and the latter being relatively resistant, were subjected to a long-term carcinogenesis experiment using our intermittent feeding protocol of PhIP in combination with a high-fat diet, which serves as a relevant risk factor that promotes the development of colon cancers. Animals were sacrificed at 60 weeks, and global gene expression analyses of normal parts of colon epithelial tissues were conducted using a high density oligonucleotide microarray to elucidate the differential gene expression profile (environment) in normal colonic regions between F344 and ACI strains. Of 8799 entries on the RatU34A array, 74 genes exhibited 3-fold or greater variation. A subset of genes encoding ribosomal RNAs and proteins were highly preferentially expressed in the F344 strain. In addition, genes encoding fatty acid binding proteins and the peroxisome membrane protein 70 appeared up regulated in the susceptible F344 strain. In the ACI strain, a mismatch repair gene, Msh2, was preferentially expressed, at approximately 20-fold the F344 level, along with a gene encoding a detoxification enzyme, catechol-O methyltransferase. The combined effects of the repertoire of these differentially expressed genes in normal colon epithelial tissues may account for the distinct susceptibilities of F344 and ACI strains to colon carcinogenesis. PMID- 12901792 TI - Repeated hepatocyte injury promotes hepatic tumorigenesis in hepatitis C virus transgenic mice. AB - Although hepatitis C virus (HCV) is a well-known causative agent of hepatocellular carcinoma (HCC), the mechanism by which HCV induces HCC remains obscure. To elucidate the role of HCV in hepatocarcinogenesis, a model of hepatocyte injury was established using HCV core transgenic mice, which were developed using C57BL/6 mice transfected with the HCV core gene under control of the serum amyloid P component promoter. After 18-24 months, neither steatosis nor hepatic tumors were found in transgenic mice. The extent of hepatocyte injury and tumorigenesis were then examined in transgenic mice following repeated administration of carbon tetrachloride (CCl(4)) using various protocols (20%, 1/week; 10%, 2/week and 20%, 2/week). Serum alanine aminotransferase (ALT) levels did not differ among HCV core transgenic mice and non-transgenic littermates; however, after 40 weeks, hepatic adenomas preferentially developed in transgenic mice receiving 20% CCl(4) once weekly. Moreover, HCC was observed in transgenic mice receiving 2 weekly injections of a 20% solution of CCl(4), and was not observed in the non-transgenic control mice. In conclusion, the HCV core protein did not promote hepatic steatosis or tumor development in the absence of hepatotoxicity. However, the HCV core protein promoted adenoma and HCC development in transgenic mice following repeated CCl(4) administration. These results suggest that hepatotoxicity resulting in an increased rate of hepatocyte regeneration enhances hepatocarcinogenesis in HCV-infected livers. Furthermore, this experimental mouse model provides a valuable method with which to investigate hepatocarcinogenesis. PMID- 12901793 TI - Distinct mechanisms of site-specific oxidative DNA damage by doxorubicin in the presence of copper(II) and NADPH-cytochrome P450 reductase. AB - The anticancer mechanism of doxorubicin (DOX), an anthracycline antibiotic, is believed to involve DNA damage through topoisomerase II inhibition and free radical generation. The free radical generation may also participate in genotoxicity, as well as cardiotoxicity, in normal human cells. The present study showed that DOX generates 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxo-dG), an indicator of oxidative DNA damage, in HL-60 cells, but not in H(2)O(2)-resistant HP100 cells, suggesting the involvement of H(2)O(2) in cellular DNA damage. Since DOX has both p-quinone and p-hydroquinone residues, free radical generation can be initiated by either reduction or oxidation of DOX. To clarify whether the oxidized or reduced form is more important for DOX-induced H(2)O(2) generation, we investigated the site-specific DNA damage induced by DOX in the presence of Cu(II), in comparison with that in the presence of cytochrome P450 reductase, using (32)P-labeled DNA fragments. DOX caused DNA damage in the presence of Cu(II) or cytochrome P450 reductase. The degree of Cu(II)-mediated DNA damage, including 8-oxodG formation, was much greater than that of cytochrome P450 reductase-mediated DNA damage. DOX plus Cu(II) caused DNA damage specifically at guanine, thymine and cytosine residues, particularly at 5'-GG-3', 5'-GT-3' and 5' TG-3' sequences. Scavenger experiments suggested the involvement of reactive species generated from H(2)O(2) and Cu(I). When cytochrome P450 reductase and NADPH were used instead of Cu(II), every nucleotide was uniformly damaged, suggesting the participation of.OH. We conclude that DOX may induce carcinostatic and genotoxic effects through oxidation of its p-hydroquinone moiety by metal ion rather than through p-quinone reduction by cytochrome P450 reductase. PMID- 12901794 TI - DNA hypermethylation and histone hypoacetylation of the HLTF gene are associated with reduced expression in gastric carcinoma. AB - The SWI/SNF proteins are ATP-dependent chromatin remodeling enzymes that have been implicated in the regulation of gene expression. Recent studies have shown that members of the SWI/SNF superfamily can function as tumor suppressor genes. DNA methylation and transcriptional inactivation of the HLTF gene, which is a homologue to the SWI/SNF genes, have been observed in colon cancer. In the present study, we studied the DNA methylation status of the HLTF gene by methylation-specific PCR in 50 gastric carcinoma tissues, and seven gastric carcinoma cell lines and compared the methylation status with the levels of HLTF mRNA expression. DNA methylation of the HLTF gene was found in 25 (50%) of 50 gastric carcinomas, and levels of HLTF mRNA were associated with methylation status of HLTF (P = 0.027; Mann-Whitney U test). No correlations were found between HLTF mRNA levels and DNA methylation and T grade, N grade, tumor stage, or histological type. In corresponding non-neoplastic mucosae, DNA methylation of the HLTF gene was found in 1 (7%) of 15 samples. The methylated allele was not detected in any of 10 normal gastric mucosae from 10 healthy volunteers. Among seven gastric carcinoma cell lines, the KATO-III cell line showed loss of HLTF mRNA expression associated with DNA methylation. This loss was rectified by treatment with both Aza-2'-deoxycytidine, a demethylating agent, and trichostatin A, a histone deacetylase inhibitor. Chromatin immunoprecipitation assay revealed that the acetylation levels of histones H3 and H4 in the 5' CpG island of the HLTF gene were inversely associated with DNA methylation status. These results suggest that transcriptional inactivation of HLTF by aberrant DNA methylation and histone deacetylation may be involved in stomach carcinogenesis through down regulation of HLTF expression. PMID- 12901795 TI - Identification of genes differentially expressed in a newly isolated human metastasizing esophageal cancer cell line, T.Tn-AT1, by cDNA microarray. AB - We isolated a metastasizing human esophageal squamous cell carcinoma (SCC) cell line, T.Tn-AT1, from a parental non-metastasizing cell line, T.Tn, by in vitro selection and by use of a nude mouse orthotopic inoculation model. Then, we compared the expression profiles of 9206 genes in T.Tn-AT1 and T.Tn by cDNA microarray analysis. The gene expression profiles of T.Tn and T.Tn-AT1 were very similar, and only 34 genes showed more than 3-fold differential expression. Among the 34 genes, 29 genes were down-regulated and only 5 genes were up-regulated in T.Tn-AT1 cells. Subsequently, we confirmed the expression levels of 14 of the 34 genes in T.Tn and T.Tn-AT1 cells by means of reverse transcription-polymerase chain reaction. The expression of 8 genes (KAL1, HPGD, NDN, REG1A, CXCR4, SPOCK, DIAPH2 and AIF1) was down-regulated and that of one gene (VNN2) was up-regulated in T.Tn-AT1 cells. These 9 genes encoded proteins associated with metastatic processes, such as adhesion, migration, inflammation, proliferation, and differentiation. Thus, these genes might regulate the metastasis of esophageal SCC, and could be predictive markers for lymph node metastasis of esophageal SCC. PMID- 12901796 TI - The implication of background anthracosis in the development and progression of pulmonary adenocarcinoma. AB - In order to characterize the relationship between background anthracosis and pulmonary adenocarcinogenesis, surgically resected tissues of 66 cases of stage I pulmonary adenocarcinoma, 4 cm or less at their greatest dimension, were examined. These cases were diagnosed based on the classification of small-sized adenocarcinoma of the lung (Noguchi et al., Cancer 75, 1995). Thirteen cases were diagnosed as types A (localized bronchioloalveolar adenocarcinoma, LBAC) and B (LBAC with alveolar collapse), 40 cases as type C (LBAC with a focus of fibroblastic proliferation), 8 as type D (poorly differentiated adenocarcinoma) and 5 as types E (bronchial gland type adenocarcinoma) and F (true papillary adenocarcinoma). The 5-year survival rate of types A and B cases was 100%, while those of type C, type D and types E and F were 52%, 48% and 39%, respectively. Nuclear accumulation of abnormal p53 protein in non-replacement type adenocarcinomas (types D, E and F) was detected more frequently than that in replacement type adenocarcinomas (types A, B and C) (P < 0.05). In each case, black dusty material was extracted from tumorous lesions and non-tumorous regions and blotted onto a nitrocellulose membrane. The anthracotic index (AI) was calculated with a densitometer. AIs of non-tumorous regions in early and replacement type adenocarcinomas (types A and B) were significantly less than in relatively advanced (type C) and poorly differentiated (type D) adenocarcinomas (P < 0.05). These results indicated that adenocarcinoma developing in heavily anthracotic lungs readily progresses to an advanced stage, or that adenocarcinoma with a less favorable prognosis tends to develop in severely anthracotic lungs. PMID- 12901797 TI - Overexpression of the Wilms' tumor gene WT1 in colorectal adenocarcinoma. AB - Expression of the Wilms' tumor gene WT1 was examined in 59 cases of colorectal adenocarcinoma to examine the involvement of WT1 in tumorigenesis. Quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR) showed that WT1 mRNA was expressed in the range from 7.2 x 10(-5) to 4.9 x 10(-1) levels (WT1 expression level in K562 leukemic cells was defined as 1.0) in all (100%) of the 28 cases of colorectal adenocarcinoma examined, and that the WT1 mRNA expression levels were higher in 20 (71%) of the 28 cases compared to those of normal appearing mucosal tissues examined. Immunohistochemical analysis using an anti WT1 antibody was performed on 46 cases of colorectal adenocarcinoma (15 of the 28 cases with WT1 mRNA expression and 31 newly collected cases), and the expression of WT1 protein was detected in 41 (89%) of the 46 cases. The direct sequencing analysis of the WT1 genomic DNA showed no mutations in any of the 10 exons of the WT1 gene in any of 5 different colorectal adenocarcinomas. These results may indicate an important role of the wild-type WT1 gene in tumorigenesis of colorectal adenocarcinoma. PMID- 12901798 TI - A naturally occurring p73 mutation in a p73-p53 double-mutant lung cancer cell line encodes p73 alpha protein with a dominant-negative function. AB - p73, a close homolog of p53 tumor suppressor, induces growth arrest and apoptosis. However, its role in cancers is controversial because of the rarity of p73 mutations, lack of tumors in p73-knockout mice, and the presence of multiple isotypes, among which Delta N isotypes inhibit the function of TA isotypes. We analyzed three naturally occurring p73 mutants found in lung cancer cell lines, NCI-H1155, DMS 92 and A427. NCI-H1155 is a cell line that has a p73 mutation [p73(G264W)] in the DNA-binding domain, as well as a p53 mutation [p53(R273H)], which is frequently found in human cancers and has a "gain-of-function" characteristic. p73 alpha(G264W) not only lacks transactivation activity itself, but also suppressed the transactivation activity of the wild-type p73 alpha in a dose-dependent manner, indicating that p73 alpha(G264W) is a dominant-negative mutant. p73 alpha(G264W) failed to suppress colony formation. We tested two other mutations, p73(Del418) in DMS 92 and p73(Del603) in A427. Both mutants retained similar levels of transactivation activity and suppression of colony formation to those of wild-type p73. The biological significance of these two mutations is unclear. In NCI-H1155 cells the coexistence of mutations that abrogate the normal functions of p73 and p53 may indicate that each mutation confers an additive growth advantage upon the cells. PMID- 12901799 TI - A nonsense mutation in exon 8 of the APC gene (Arg283Ter) causes clinically variable FAP in a Malaysian Chinese family. AB - The present study was carried out to characterize the causative genetic mutation in a medium-sized Malaysian Chinese pedigree of three generations affected with familial adenomatous polyposis (FAP). Clinical data and genetic studies revealed considerable phenotypic variability in affected individuals in this family. Blood was obtained from members of the FAP-01 family and genomic DNA was extracted. Mutation screening of the adenomatous polyposis coli (APC) gene was carried out using the single strand conformation polymorphism (SSCP) technique. The possibility of exon skipping was predicted by splicing motif recognition software (ESEfinder release2.0). SSCP results showed mobility shifts in exon 8 of the APC gene which segregated with affected members of the family. Sequence analysis revealed that the affected individuals are heterozygous for a C847T transition, whilst all the unaffected family members and control individuals are homozygous C at the same position. This nucleotide substitution generates a stop codon at amino acid position 283, in place of the usual arginine (Arg283Ter). We conclude that an Arg283Ter mutation in the APC gene is causative of the FAP phenotype in this family, although there is considerable variation in the presentation of this disease among affected individuals. Computational analysis predicts that this mutation occurs within sequences that may function as splicing signals, so that the sequence change may affect normal splicing. PMID- 12901801 TI - The role of circulating IL-8 and VEGF protein in the progression of gastric cancer. AB - Both vascular endothelial growth factor (VEGF) and interleukin 8 (IL-8) play an important role in the progression of gastric cancer (GC). In this study, we investigated whether circulating levels of VEGF or IL-8 in drainage veins of GC patients were correlated with any clinicopathological factors. Thirty-seven patients with primary GC who underwent gastrectomy at our department between 1999 and 2002 were analyzed. Blood samples were drawn from a peripheral vein just before surgery and from a drainage vein immediately after laparotomy. Plasma VEGF levels were significantly higher than those in 10 healthy controls. There was no correlation between VEGF levels in drainage veins and any clinicopathological variable, whereas there was a significant relationship in the case of VEGF levels in peripheral veins; the levels were higher in patients with venous invasion. We found a significant relationship between IL-8 levels in drainage veins and both tumor size and lymph node metastasis, whereas no significant relationship between IL-8 levels in peripheral veins and any variable was found. There was no correlation between VEGF and IL-8 levels in drainage veins. Large tumors, deeply invasive tumors, lymph node involvement, venous invasion and high IL-8 levels in drainage veins were all significantly associated with shorter disease-free survival, although multivariate analysis revealed that lymph node involvement was the only independent prognostic factor. In conclusion, the measurement of IL-8 levels in drainage veins of GC patients may reflect production mainly by the primary lesion and is valuable as an indicator of risk for recurrent disease. PMID- 12901800 TI - Long-term follow-up of patients with unresectable locally advanced non-small cell lung cancer treated with chemoradiotherapy: a retrospective analysis of the data from the Japan Clinical Oncology Group trials (JCOG0003A). AB - To clarify the long-term survival data and factors that are correlated with survival outcome of unresectable locally advanced non-small cell lung cancer (NSCLC) following chemoradiotherapy, we analyzed patients who entered the Japan Clinical Oncology Group (JCOG) clinical trials for unresectable locally advanced NSCLC. Between October 1989 and August 1997, 240 patients (male 207, female 33; PS (performance status) 0 58, PS 1 172, PS 2 9, unknown 1; stage IIB 2, IIIA 62, IIIB 175, unknown 1) entered the 6 trials. All patients received chemotherapy and radiotherapy. The associations between survival outcome and treatment-related factors were analyzed using Cox regression analysis. Median survival times and 5 year survival rates in the trials were 11.9-19.7 months and 0-17.6%, respectively. Median survival time was 16.1 months and the 5- and 7-year survival rates of all 240 patients were 14.4% and 12.0%, respectively. No deaths were observed 7 years after initiation of the treatment or later. For stage IIIA and IIIB patients, the 5-year survival rates were 16.3% and 13.4%, respectively. Node status and age were significantly associated with survival, but no factors of the treatment were associated with survival of patients with unresectable locally advanced NSCLC. The present retrospective analysis showed that approximately 12% of patients with unresectable locally advanced NSCLC could be cured by various chemoradiotherapy regimens. PMID- 12901802 TI - Time course of carcinoembryonic antigen after resection of lung cancer: a predictor of recurrence. AB - We investigated whether the early postoperative time course of carcinoembryonic antigen (CEA) level after resection of lung cancer could be used to predict patients' prognosis. Fifty-three lung cancer patients were included in this study. Postoperative serum CEA levels were calculated by means of non-linear least-squares fitting to the equation C(t) = (C(0)-C(p))exp(-kt) + C(p), where C(t) is postoperative CEA level, t is days after surgery, C(0) is CEA level at postoperative time 0, C(p) is CEA level at plateau, and k is the rate constant of elimination. Postoperative CEA production (P(p)) was calculated as C(p) multiplied by k. C(p) and P(p) represent the numbers of residual tumor cells after surgery. More residual tumor cells yield higher values of C(p) and P(p), and result in earlier recurrence. RESULTS: Kinetic parameters could be obtained for 30 patients whose preoperative CEA levels were sufficiently elevated. Cutoff levels as predictors for recurrence were 1.1 ng/ml for C(p) and 0.9 ng/ml/day for P(p). The accuracy of prediction of recurrence using these cutoff levels was 79% with C(p) and 89% with P(p). A very poor prognosis was observed for patients with P(p) over 0.9 ng/ml/day. CONCLUSION: Analysis of the time course of changes in CEA levels after resection of lung cancer appears to be useful for predicting patient prognosis. C(p) and P(p) are very precise predictors of recurrence. PMID- 12901803 TI - Adenovirus-mediated gene transduction of truncated I kappa B alpha enhances radiosensitivity in human colon cancer cells. AB - Nuclear factor kappa B (NF-kappa B) is a transcription factor that is known to regulate apoptosis when cells are exposed to DNA-damaging agents such as ionizing radiation and cytotoxic drugs. We sought to determine if inhibition of NF-kappa B could enhance radiosensitivity in human colon cancer cells in vitro and in vivo. To inhibit NF-kappa B activation specifically, we constructed a recombinant adenovirus vector expressing a truncated form of the inhibitor protein I kappa B alpha (I kappa B alpha Delta N) that lacks the phosphorylation sites essential for activation of NF-kappa B, and transfected two human colon cancer cell lines (HT29 and HCT15) with this vector. In vitro colony-forming assays revealed that the overexpression of the stable I kappa B alpha by AxI kappa B alpha Delta N infection significantly suppressed cell growth after irradiation in both cell lines as compared to infection with a control vector, AxLacZ. Treatment with AxI kappa B alpha Delta N and irradiation successfully inhibited the growth of HT29 xenografted subcutaneous tumors in nude mice with an 83.8% volume reduction on day 38 as compared to the untreated tumors. Furthermore, it was demonstrated that apoptosis was increased by adenovirus-mediated gene transduction of I kappa B alpha Delta N in vitro and in vivo. These results indicated that inhibition of NF kappa B could enhance radiosensitivity through an increase in radiation-induced apoptosis. We believe that radio-gene therapy using adenovirus-mediated gene transduction of I kappa B alpha Delta N could be an attractive candidate as a treatment strategy for colorectal cancer. PMID- 12901804 TI - Management of chronic parotitis: a review. AB - Chronic parotitis is an insidious inflammatory disorder, which is characterized by a recurrent, often painful, swelling of the gland or glands. Sialography remains the investigation of choice. Diagnostic siladenoscopy may complement or supersede sialography as the diagnostic procedure of choice, as more experience is gained in this technique and its use becomes more widespread. The management options are conservative or surgical, but controversy exists regarding the timing and method of surgical intervention. The surgical methods that have been described include injection of methyl violet - a sclerosing agent, ductal ligation, ductoplasty, tympanic neurectomy, and parotidectomy. Parotidectomy is considered the ultimate surgical option but because of its significant morbidity the exact technique and procedure that should be used remains contentious. PMID- 12901805 TI - Evaluation of the HiFocus electrode array with positioner in human temporal bones. AB - The aim of the study presented was to assess the insertion mode and possible intracochlear trauma after implantation of the HiFocus electrode with positioner in human temporal bones. The study was performed in five freshly frozen temporal bones. The position of electrodes was evaluated using conventional X-ray analysis, rotational tomography and histomorphological analysis. Insertion of the HiFocus electrode with positioner resulted in considerable trauma to fine cochlear structures including fracture of the osseous spiral lamina, dislocation of the electrode array from the scala tympani into the scala vestibuli and fracture of the modiolus close to the cochleostomy. The implication of the results regarding clinical outcome will be discussed. PMID- 12901806 TI - Live recordings of sound levels during the use of powered instruments in ENT surgery. AB - Otolaryngology is one of the surgical specialities employing high-powered instruments and this study was designed in order to establish whether sound levels at work conform to HSE guidelines. No study to date has measured intra operative noise levels in ENT operation theatres. A prospective observational study was therefore performed. Sound levels were measured during the use of bone drills for mastoid surgery and microdebriders for endoscopic sinus surgery. A SLM/IS Acos Class I sound level meter calibrated to BS 1259 was employed. A spectrum analysis of drill-generated noise was measured using a calibrated B and K precision sound level meter. Sound levels emitted varied as follows: Large burrs- 72.4 dB (A), medium size; 71.2 dB (A), small sized burrs- 68.8 dB (A) (all values for cutting burrs) and 60.8 dB (A) for diamond burs. With microdebriders, the average sound level was 60.1 dBA. Spectrum analysis revealed that the maximum intensity was at 3.15 kHz, followed by 4 kHz, while the least sound was produced at 40 kHz and 31.5 kHz. Essentially sound produced by drills was between 1.6 kHz and 6.3 kHz. Powered instruments used in ENT surgery are safe and pose no occupational hazard. PMID- 12901807 TI - Consent processes in common nose and throat procedures. AB - The NHS Plan identified the need for changes in consent practice, and led to the issuing of a model consent policy and standard consent forms for use in the NHS. The aim of this study was to determine current consent practices for common rhinology and laryngology procedures. A telephone survey was conducted of 40 otolaryngology Senior House Officers (SHOs) across England and Wales, asking about local consent procedure, and the specific complications discussed before common operations. The responsibility for routine consenting belonged to SHOs in 95 per cent of departments. Model NHS consent forms were used in 72.5 per cent and information sheets given to patients in 25 per cent of departments. The specific operative risks mentioned to patients by the SHOs showed great variability. The provision of standardized consent protocols for each operation, together with information sheets and the model NHS forms, could aid juniors involved in consent and prove beneficial to our patients. PMID- 12901808 TI - Laser posterior ventriculocordectomy with partial arytenoidectomy for the treatment of bilateral vocal fold immobility. AB - Bilateral vocal fold immobility in adduction usually creates severe dyspnoea. Many surgical procedures have been established to improve the airway insufficiency in affected patients. Over the last six years 22 patients with bilateral vocal fold immobility in our department have undergone CO(2) laser posterior ventriculocordectomy with partial arytenoidectomy (PVCPA). None had dyspnoea or a disturbance in the immediate post-operative period or during follow up, which ranged from five months to six years. Only one patient required a second procedure. Laser PVCPA appears to be an effective and reliable method for the treatment of bilateral vocal fold immobility in selected patients. PMID- 12901809 TI - Botulinum toxin injection for failed tracheo-oesophageal voice in laryngectomees: the Sunderland experience. AB - Spasm of the pharyngo-oesophageal segment is one of the important causes of tracheo-oesophageal voice failure. Traditionally it has been managed by either prolonged speech therapy, surgical pharyngeal myotomy or pharyngeal plexus neurectomy with varying degrees of success. Botulinum neurotoxin has been found to be effective in relieving pharyngo-oesophageal segment spasm. Since 1995, we have used botulinum toxin injection on 10 laryngectomees with either aphonia or hypertonicity due to pharyngo-oesophageal segment spasm. Early results were analysed by the Sunderland Surgical Voice Restoration Rating scale. Seven of the 10 patients, who were previously completely aphonic, developed voice following this therapy and are using their valve choice as their only method of communication. Out of the three patients who were treated for hypertonic voice, two did derive some benefit from the procedure. One patient developed a hypotonic voice, which lasted for a few months. PMID- 12901810 TI - Lip injury prevention during tonsillectomy. AB - Patients commonly complain of discomfort at the angle of the mouth following tonsillectomy. This can be attributed to trauma sustained during the procedure. A prospective, patient-blinded, controlled trial of 34 tonsillectomies was performed using a protective, plastic oral guard. A blinded observer recorded postoperative pain and clinically evident trauma to the angle of the mouth. The patient group in whom the guard was used had a significantly lower incidence of labial trauma (p<0.002) and complained of pain less frequently (p<0.002) when compared with a control group in whom no guard was used. The use of a protective lip guard during tonsillectomy is recommended. PMID- 12901811 TI - New surgical adjuncts in the treatment of rhinophyma: the microdebrider and FloSeal. AB - Rhinophyma is considered to be the final stage of rosacea and has an unknown aetiology. We present two new surgical adjuncts for the treatment of this condition. The microdebrider is easy to use and allows precise contouring of the nose. It shares the same underlying principle as free-hand scalpel shaving but outperforms this method in key areas. We also describe a novel use for FloSeal((R)), a gelatin-thrombin co-mixture, utilizing its haemostatic properties to address bleeding from the re-shaped denuded nose. PMID- 12901813 TI - Supralabyrinthine approach to petrosal cholesteatoma. AB - Petrosal cholesteatomas are rare lesions, which may be congenital or acquired in nature. We report an exceptional case occurring in a seven-year old girl who presented with a unilateral conductive hearing loss, despite normal tympanic membrane appearance. Early diagnosis was facilitated by computed tomography (CT) scanning. Although this case satisfied the criteria for congenital cholesteatoma, it is likely that the petrous apex was secondarily involved. Complete cholesteatoma removal was accomplished using a transtemporal supralabyrinthine approach, which allowed for hearing preservation, while avoiding the morbidity associated with a craniotomy. The present case constitutes the youngest case of petrosal cholesteatoma reported. We suggest that a lower threshold for the use of CT scanning in unilateral conductive hearing loss may allow for the earlier detection of more cases of petrosal cholesteatomas, as well as facilitating their removal using more limited approaches associated with less morbidity. PMID- 12901812 TI - Superior semicircular canal dehiscence simulating otosclerosis. AB - This is a report of a patient with an air-bone gap, thought 10 years ago to be a conductive hearing loss due to otosclerosis and treated with a stapedectomy. It now transpires that the patient actually had a conductive hearing gain due to superior semicircular canal dehiscence. In retrospect for as long as he could remember the patient had experienced cochlear hypersensitivity to bone-conducted sounds so that he could hear his own heart beat and joints move, as well as a tuning fork placed at his ankle. He also had vestibular hypersensitivity to air conducted sounds with sound-induced eye movements (Tullio phenomenon), pressure induced nystagmus and low-threshold, high-amplitude vestibular-evoked myogenic potentials. Furthermore some of his acoustic reflexes were preserved even after stapedectomy and two revisions. This case shows that if acoustic reflexes are preserved in a patient with an air-bone gap then the patient needs to be checked for sound- and pressure-induced nystagmus and needs to have vestibular-evoked myogenic potential testing. If there is sound- or pressure-induced nystagmus and if the vestibular-evoked myogenic potentials are also preserved, the problem is most likely in the floor of the middle fossa and not in the middle ear, and the patient needs a high-resolution spiral computed tomography (CT) of the temporal bones to show this. PMID- 12901814 TI - Sphenoid sinus mucocele: late complication of Sphenoidotomy. AB - Sphenoid sinus mucocele is an infrequent but well-recognized entity in sinus pathology. The symptoms produced by it are related to the anatomical surroundings of the sphenoid sinus. We describe a case of sphenoid sinus mucocele presenting with ipsilateral oculomotor nerve palsy 10 years after endoscopic sphenoidotomy. The patient underwent emergency endoscopic decompression of the sphenoid sinus with marsupialization of the mucocele, resulting in immediate remission of his symptoms. We conclude that the otorhinolaryngologist should be aware of such a potential complication when counselling the patient prior to endoscopic sinus surgery. PMID- 12901815 TI - Lymphangioma of the sphenoid sinus. AB - Lymphangiomas are rare benign lymphatic tumours found predominantly in the head and neck region. A case of a cavernous lymphangioma isolated to the sphenoid sinus is described. The authors emphasize the extreme rarity of the isolated sphenoid lymphangioma, as to their knowledge this is the first report in the English literature. PMID- 12901816 TI - Unusual complication of ingestion of a foreign body. AB - Foreign bodies of the upper aerodigestive tract are common problems dealt with by the otolaryngologist. Among all foreign bodies in the oesophagus, an open safety pin still presents a challenge for the ENT specialists because of its propensity to pierce the oesophagus and surrounding structures. We present an interesting case of a long-standing foreign body i.e. an open safety pin, which, after piercing the hypopharynx, caused fatal common carotid artery rupture. PMID- 12901817 TI - Fish bone as a foreign body. AB - Fish bones are one of the most common foreign bodies in the upper aero-digestive tract. The use of plain X-ray in identifying fish bones has questionable value. We believe that the knowledge of the type of the fish ingested improves the diagnostic value of the neck X-rays. This study was designed to evaluate the relative radio-densities of the bones of commonly eaten fish in the UK. Twenty three species of fish were studied and their bones were grouped into three depending upon their ratio-densities. This information is expected to be useful in identifying fish bones while reading plain radiography. PMID- 12901818 TI - Angiolymphoid hyperplasia with eosinophilia arising from the facial artery. AB - We report on a rare case of angiolymphoid hyperplasia with eosinophilia (ALHE) arising from the facial artery, which was pre-operatively diagnosed as an aneurysm. ALHE is a rare lesion, the precise nature of which remains controversial: it is still unclear whether ALHE is a tumour or an inflammatory disease or whether it can be separated from Kimura's disease based on its pathological characteristics. ALHE rarely occurs in muscular arteries, and this is the first description of ALHE arising from the facial artery. Angiography vividly revealed the pathological structure of the lesion in this case, which prompted us to advocate angiography for diagnosing ALHE arising from muscular arteries. Complete dissection and cryotherapy proved to be effective treatment, although complete management of any therapeutic protocol should avoid recurrence of the lesion, whatever specific treatments are selected. PMID- 12901819 TI - Dual ectopic thyroid: report of a case. AB - Dual ectopic thyroid is very rare. We report a case of dual ectopic thyroid in the lingual and infrahyoid areas in a 20-year-old female patient with no thyroid gland in its normal anatomical location. On physical examination, there was a 7 x 5 cm anterior midline neck swelling just below the hyoid bone and a 2 x 2 cm mass in the base of the tongue. Triiodothyronine (T(3)), thyroxine (T(4)), and thyroid stimulating hormone (TSH) levels were normal. A thyroid scan with technetium-99m sodium pertechnate confirmed dual ectopic thyroid with no iodine uptake in the normal anatomical location of the thyroid gland. The infrahyoid ectopic thyroid was surgically removed for cosmetic reasons, and the lingual thyroid, which was symptomatic, was left untouched. The importance of thyroid scanning in the evaluation of anterior midline neck swellings and treatment options are discussed. PMID- 12901820 TI - Epistaxis as a rare presenting feature of glomus tympanicum. AB - Glomus tumours are the most common primary neoplasms of the middle ear, typically benign and slowly progressive. Pulsatile tinnitus and ipsilateral hearing loss are the most common symptoms at presentation by far; otalgia, aural fullness and otorrhoea being less frequent. A case of primary glomus tympanicum presenting with recurrent epistaxis, previously unreported in the literature, is described and associated imaging presented. PMID- 12901821 TI - Anomalies of the auditory organ in trisomy 18 syndrome: human temporal bone histopathological study. AB - The purpose of this study was to define the histopathological changes in the temporal bone of a fetus with trisomy 18 syndrome, a stillborn due to perosplanchnia. Several anomalies were found including malformation of the auditory ossicles, residual mesenchyme in the middle ear, aberrant tensor tympani muscle, absence of stapedial tendon, aberrant lateral ampullary nerve and wide endolymphatic sinus. The incus body was deformed and separated from the long process by connective tissue and monocrural stapes was noted in the right ear. Three-dimensional reconstruction images provided a clear view of the auditory ossicle malformation. The abnormal findings in our case indicate that ear anomalies in this syndrome might be derived from the component around the first and second branchial arches. PMID- 12901822 TI - Pigmented vagal paraganglioma. AB - Paragangliomas are uncommon and those arising from the vagal trunk are rarer. Pigmented extra-adrenal paragangliomas are still rarer and reported sites of occurrence are the uterus, spine, retroperitoneum, bladder, mediastinum and orbit. The presence of abundant pigment in a cervical paraganglion has not been reported previously. We report one such unusual case of pigmented vagal paraganglioma that arose from the vagal trunk below the nodose ganglion, had massive central necrosis and showed hypovascularity on angiography. The unusual features and difficulties in the diagnosis of such cases are discussed. PMID- 12901823 TI - [Application of St George's respiratory questionnaire in evaluating the life quality of Chinese patients with chronic obstructive pulmonary disease]. AB - OBJECTIVE: To study the value of St George's respiratory questionnaire (SGRQ) in evaluating the life quality of Chinese patients with COPD. METHODS: The SGRQ score and spirometry were collected from 30 patients with COPD. The validity was documented by performing correlation analysis, and Pearson's correlation coefficients were calculated. Nineteen patients were reassessed after three months' treatment with inhaled glucocorticoids. Paired t test was performed to evaluate the sensitivity of SGRQ to changes of disease severity. RESULTS: The correlation coefficients between the total score and the symptoms section score, the activity section score, and the impacts section score of SGRQ were 0.61, 0.79 and 0.97, respectively. The symptoms section score, the activity section score, the impacts section score, and the total score were significantly correlated with FEV(1)% pred, and with similar coefficients (-0.4 - -0.5, P < 0.05). The result of paired t test showed that the changes of SGRQ between the two assessments were significant (P < 0.001) and consistent with changes of clinical symptoms and spirometry. SGRQ was more sensitive than spirometry in evaluating the changes of disease severity. CONCLUSION: SGRQ was a valid, sensitive and feasible method in measuring the quality of life in Chinese patients with COPD. Large-scale application in China needs to be carried out. PMID- 12901824 TI - [Study on the molecular mechanism of transferable multiple-antibiotic resistance in extended-spectrum beta-lactamase-producing clinical isolates]. AB - OBJECTIVE: To investigate the molecular mechanism of transferable multiple antibiotic resistance in extended-spectrum beta-lactamases (ESBLs) producing isolates. METHODS: Antibiotics susceptibility was tested by E-test method, and multi-resistance plasmids were screened and isolated by extracting transformant plasmids. Inserted gene Cassettes of class 1 integron were amplified and analyzed by polymerase chain reaction (PCR) and DNA sequencing. RESULTS: Eight of the nine ESBL-producing plasmids were found to comprise class 1 integron sequence, of them 7 harbored 1 or 2 antibiotic resistant gene cassettes which encoding resistance to aminoglycosides (aacA4, aadA2 or aadA5), trimethoprim (dhfrA12 or dfrA17), rifampicin (arr-3) and chloramphenicol (cmlA6). The function of these gene cassettes corresponded to the resistance profiles of their electro-transformants. CONCLUSION: Multi-resistance gene cassettes located on plasmids and mediated by class 1 integron may play an important role in causing the development and dissemination of multiple-antibiotic resistance in ESBL-producing clinical isolates. PMID- 12901825 TI - [The relationship between polymorphism of angiotensin converting enzyme gene and cough caused by angiotensin converting enzyme inhibitors]. AB - OBJECTIVE: To investigate the relationship between the polymorphism of angiotensin converting enzyme (ACE) gene and ACE inhibitor (ACEI)-induced cough in Chinese elderly with essential hypertension. METHODS: ACE I/D polymorphism was performed on DNA samples from patients using the polymerase chain reaction (PCR) followed by agarose gel electrophoresis. Serum ACE levels were measured using a colorimetric assay. RESULTS: The frequencies of ACE II genotype were 40% in patients with cough and 20% in those without cough, respectively (P < 0.05). The frequencies of I allele were 60% in patients with cough and 41% in those without cough (P < 0.01); The serum ACE level was highest in the DD genotype, followed by the ID genotype and the II genotype. The ACE level in patients with cough was significantly lower than that in patients without cough (P < 0.001). The sensitivity and specificity were 81% and 78% respectively when the serum ACE levels were used to predict ACEI-induced cough. CONCLUSION: ACEI-induced cough was related to the serum ACE level and ACE gene polymorphism in Chinese elderly with essential hypertension. PMID- 12901826 TI - [The effects of erythromycin on the secretion of tumor necrosis factor-alpha and transforming growth factor-beta1 and expression of connexin 43 in human pleural mesothelial cells]. AB - OBJECTIVES: To investigate the effects of erythromycin on secretion of tumor necrosis factor-alpha (TNF-alpha) and transforming growth factor-beta(1) (TGF beta(1)), and the level of connexin 43 in human pleural mesothelial cells (HPMC), and to explore the mechanism of pleurodesis. METHODS: HPMC was incubated with erythromycin at concentrations of 100 mg/L or 25 mg/L. The levels of TNF-alpha and TGF-beta(1) in the supernatants were measured by ELISA method, and levels of connexin 43 were detected by Western blot. RESULTS: The secretion of TNF-alpha by HMPC increased after incubation with 100 mg/L erythromycin for 3 or 5 days, and the secretion of TGF-beta(1) increased markedly after incubation with lower or higher concentrations of erythromycin. The levels of connexin 43 in HPMC decreased after stimulation with 100 mg/L erythromycin, but no relationship was observed between the levels and the stimulation time. CONCLUSIONS: HPMC incubated with erythromycin showed increased secretion of TNF-alpha and TGF-beta(1), which may be one of the mechanisms for erythromycin pleurodesis. Erythromycin decreased the level of connexin 43 in HPMC, and this effect may be a response of the cells to the stimulus. A high concentration of erythromycin is suggested for clinical pleurodesis based on the results from this study. PMID- 12901827 TI - [Heparanase and vascular endothelial growth factor expression in non-small-cell lung cancer]. AB - OBJECTIVE: To study the clinicopathological significance of heparanase and VEGF expression in NSCLC. METHODS: Eighty-five lung samples were studied. Each sample was divided into two parts, one used for heparanase mRNA detection by reverse transcription PCR and the other for VEGF detection by immunohistochemistry. RESULTS: (1) The expressions of heparanase mRNA and VEGF were higher in NSCLC than in benign pulmonary diseases (P < 0.05). (2) The expression of heparanase mRNA was higher in cases with lymph-node invasion, metastasis, stage III and IV diseases, low-differentiation, and adenocarcinoma, as compared to cases without lymph-node invasion and metastasis, with stage I and II diseases, higher and moderate differentiation, and squamous cell cancer (P < 0.05). Its expression was higher in tumors larger than 5 cm in size (P < 0.05). (3) The expression of VEGF was higher in cases with lymph-node invasion, metastasis, and stage III and IV disease, as compared to cases without lymph-node invasion and metastasis, and with stage I and II diseases (P < 0.05). (4) There was no significant correlation between heparanase and VEGF expression (P > 0.05). CONCLUSIONS: Heparanase and VEGF are associated with NSCLC invasion and metastasis, and may be used to evaluate NSCLC metastasis status. Heparanase and VEGF promote NSCLC invasion and metastasis by independent mechanisms. Detection of these two markers may improve the sensitivity and specificity of the measurement of NSCLC metastatic potential. PMID- 12901828 TI - [Nitric oxide production and expression of cytokines by macrophages infected by M. tuberculosis H(37)R(v)]. AB - OBJECTIVE: To study nitric oxide (NO) production and cytokine expression by macrophages infected by M. tuberculosis H(37)R(v), and to compare the difference between dead and live M. tuberculosis in the induction of immune responses, and thus to show if dead bacteria could be a possible candidate for new vaccines. METHODS: Reverse transcription-polymerase chain reaction (RT-PCR) and ELISA were used to measure the production of NO and cytokines in macrophages infected by H(37)R(v). RESULTS: Macrophages infected by viable M. tuberculosis produced more NO, IL-1, IL-12, IL-18, TNF-alpha and inducible nitric oxide synthases (iNOS), as compared with macrophages infected by dead bacteria. The number of bacteria was also an important factor determining the production of NO and cytokines. CONCLUSIONS: Viable M. tuberculosis H(37)R(v) can induce the activation of macrophages and the production of more NO and cytokines which play important roles in the host immune response. Heat-killed M. tuberculosis H(37)R(v) failed to induce activation of macrophages and the production of NO and cytokines, which makes it unlikely to be a candidate for vaccine development. PMID- 12901829 TI - [Effects of vaccae on airway contraction and inflammation in asthmatic guinea pigs]. AB - OBJECTIVE: To study the effects of Mycobacterium vaccae (M. vaccae) on the lung function, airway hyperresponsiveness and airway inflammation in a guinea pig model of asthma. METHODS: Asthma was induced with ovalbumin (OVA) in 71 guinea pigs. The changes of lung resistance (R(L)) and dynamic lung compliance (C(dyn)), the accumulation of inflammatory cells in bronchoalveolar lavage fluids (BALF), and the contraction reaction curve of tracheal smooth muscles to carbachol in vitro were determined. RESULTS: Pretreatment with M. vaccae as a single dosage by intramuscular injection (i.m) inhibited the early phase of asthma in the sensitized guinea pigs in a dose-dependent manner. R(L) (1 min to 15 min) increased 46.4% in the 2.5 micro g group, 29.6% in the 7.5 micro g group, and 20.8% in the 22.5 micro g group, which were significantly lower than that in the model control group (95.3%) (P < 0.05 approximately 0.01), respectively. The mean decrease of C(dyn) was 26.8% in the 2.5 micro g group, 23.5% in the 7.5 micro g group, and 21.5% in the 22.5 micro g group, which were significantly lower than that in the model control group (38.7%) (P < 0.05), respectively. Pretreatment with M. vaccae in a dosage of 2.5 micro g, 7.5 micro g and 22.5 micro g (per guinea pig) i.m also inhibited the late phase of asthma. The total white blood cells in BALF was (16.2 +/- 3.2) x 10(8)/L in the 2.5 micro g group, (14.6 +/- 3.4) x 10(8)/L in the 7.5 micro g group, and (15.4 +/- 2.5) x 10(8)/L in the 22.5 micro g group, which were significantly lower than that in the model control group (22.3 +/- 2.2) x 10(8)/L (P < 0.01 approximately 0.001), respectively. BAL eosinophil count was (11.6 +/- 3.0) x 10(8)/L in the 2.5 micro g group, (6.0 +/- 1.5) x 10(8)/L in the 7.5 micro g group, and (2.2 +/- 1.7) x 10(8)/L in the 22.5 micro g group, which were significantly lower than that in the model control group (15.9 +/- 5.2) x 10(8)/L (P < 0.01 approximately 0.001), respectively. M. vaccae significantly inhibited carbachol-induced airway hyperresponsiveness in the tracheal smooth muscle (TSM) from the sensitized guinea pigs. EC(50) (the concentration of carbachol required to increase TSM contraction value by 50% from the baseline) was 0.140 micro mol/L in the 2.5 micro g group, 0.179 micro mol/L in the 7.5 micro g group and 0.236 micro mol/L in the 22.5 micro g group, which were higher than that in the model control group (0.038 micro mol/L). CONCLUSIONS: M. vaccae was found to improve lung function, inhibit airway inflammation and airway hyperresponsiveness in this animal model, which supports its application in the treatment of asthma. PMID- 12901830 TI - [Radiological features of primary pulmonary non-Hodgkin lymphoma: report of three cases]. AB - OBJECTIVE: To describe the radiographic and CT findings of primary pulmonary non Hodgkin lymphoma (PPL),and to evaluate percutanous transthoracic needle biopsy (PTNB) in the diagnosis of PPL. METHODS: Chest radiographs and CT scans of three patients with histologically proven PPL were reviewed. The diagnosis of PPL was confirmed histologically with specimens obtained by means of PTNB. RESULTS: Consolidation with air bronchograms on the chest X-ray films and CT scans was found in all cases. Multiple ill-defined nodules of various size in both lungs and a mass with air bronchogram in the right lower lobe on the CT scan were present in one case. Ground-glass opacity or reticular lesions were present in both lung fields. Hilar/mediastinal adenopathy and pericardial/pleural effusion were not features of these cases. PTNB was diagnostic in all three cases. No complications occurred during these procedures. CONCLUSIONS: PPL has some specific imaging features. Radiographic and CT findings are helpful in the diagnosis of primary pulmonary malignant lymphoma. A specific diagnosis can be obtained by means of PTNB. PMID- 12901831 TI - [Video-assisted thoracoscopic surgery for spontaneous pneumothorax]. AB - OBJECTIVE: To explore the operational procedures, indications, and the long-term results of video-assisted thoracoscopic surgery (VATS) in the treatment of spontaneous pneumothorax. METHODS: One hundred seventy three cases of spontaneous pneumothorax treated with VATS in our hospital between 1995 and 2001 were analyzed. Primary spontaneous pneumothorax (PSP) was found in 157 cases and secondary spontaneous pneumothorax (SSP) in 16 cases. In patients with PSP, pneumothorax occurred for the first time in 65 (41.4%) cases, and recurred at least once in 92 (58.6%) cases. Among them three patients underwent simultaneous bilateral VATS for simultaneous bilateral pneumothorax, and 6 underwent two-stage bilateral VATS. In patients with SSP, 6 had recurrent pneumothorax. RESULTS: According to the Vanderschueren's classification, no abnormality was found in 34 (19.7%) cases, pleuro-pulmonary adhesions in 41 (23.7%) cases and bullae in 98 (56.7%) cases. Pleurodesis and stapling under Endo-cutter were the major operational procedures used in these cases. The median follow-up time was 53 months. The long-term recurrence rate was 1.8%. Complications related to surgery occurred in 3.0% of the cases, which included subcutaneous emphysema, localized pleural effusion and prolonged air leakage. CONCLUSION: VATS is a safe and effective treatment for both initial and recurrent spontaneous pneumothorax, and therefore it is becoming the primary treatment modality for this disease. PMID- 12901832 TI - Protofibrils of amyloid beta-protein inhibit specific K+ currents in neocortical cultures. AB - Protofibrils (PFs) are recently described intermediate assemblies formed during the fibrillogenesis of amyloidogenic proteins and may play an important pathogenic role in Parkinson's and Alzheimer's disease (AD). Here we show for the first time that amyloid beta-protein (Abeta) attenuation of specific K(+) currents is dependent on the aggregation state; PFs inhibit K(+) currents, whereas low-molecular-weight assemblies have no effect. Using patch clamp analysis in whole cell current-clamp mode, we showed that at low nanomolar concentrations Abeta(1-42) PFs induce reversible, Ca(2+)-dependent increases in spontaneous action potentials and membrane depolarizations. The low nanomolar PF concentrations used, the instantaneous responses observed, and the reversibility of the effect all suggest that PFs may bind to specific channels or membrane proteins. Switching to voltage-clamp mode, we found that PFs at 1-2 microM can inhibit specifically the 4AP-sensitive K(+) currents, A-type and D-type, but not other outward or inward rectifying K(+) channels. Finally, we show that a consequence of PF-induced membrane activity is an increase in intracellular Ca(2+) spikes that are dependent on synaptic connections in the neural network formed in culture. Our data strongly support the concept that PFs can induce subtle synaptic alterations that may underlie early symptoms of AD. PMID- 12901833 TI - Identification of a third Protein 4.1 tumor suppressor, Protein 4.1R, in meningioma pathogenesis. AB - Meningiomas are common central nervous system tumors; however, the mechanisms underlying their pathogenesis are largely undefined. In this report, we demonstrate that a third Protein 4.1 family member, Protein 4.1R, functions as a meningioma tumor suppressor. We observed loss of Protein 4.1R expression in two meningioma cell lines (IOMM-Lee, CH157-MN) by Western blotting as well as in 6 of 15 sporadic meningiomas by immunohistochemistry and fluorescence in situ hybridization. In support of a meningioma tumor suppressor function, Protein 4.1R overexpression resulted in reduced IOMM-Lee and CH157-MN cell proliferation. Similar to the Protein 4.1B and merlin tumor suppressors, Protein 4.1R membrane localization increased significantly under conditions of growth arrest in vitro. Lastly, we show that Protein 4.1R interacted with a subset of merlin/Protein 4.1B interactors including CD44 and betaII-spectrin. Collectively, these results suggest that Protein 4.1R functions as an important tumor suppressor in the molecular pathogenesis of meningioma. PMID- 12901834 TI - Kindling alters entorhinal cortex-hippocampal interaction by increased efficacy of presynaptic GABA(B) autoreceptors in layer III of the entorhinal cortex. AB - We studied the effect of kindling, a model of temporal lobe epilepsy, on the frequency-dependent information transfer from the entorhinal cortex to the hippocampus in vitro. In control rats repetitive synaptic activation of layer III projection cells resulted in a frequency dependent depression of the synaptic transfer of action potentials to the hippocampus. One-to-two-days after kindling this effect was strongly reduced. Although no substantial change in synaptic inhibition upon single electrical stimulation was detected in kindled rats, there was a significant depression in the prolonged inhibition following high frequency stimulation. In kindled animals, paired-pulse depression (PPD) of stimulus-evoked IPSCs in layer III neurons was significantly stronger than in control rats. The increase of PPD is most likely caused by an increased presynaptic GABA(B) receptor-mediated autoinhibition. In kindled animals activation of presynaptic GABA(B) receptors by baclofen (10 microM) suppressed monosynaptic IPSCs significantly more than in control rats. In contrast, activation of postsynaptic GABA(B) receptors by baclofen was accompanied by comparable changes of the membrane conductance in both animal groups. Thus, in kindled animals activation of the layer III-CA1 pathway is facilitated by an increased GABA(B) receptor mediated autoinhibition leading to an enhanced activation of the monosynaptic EC CA1 pathway. PMID- 12901835 TI - Mitochondrial dysfunction due to mutant copper/zinc superoxide dismutase associated with amyotrophic lateral sclerosis is reversed by N-acetylcysteine. AB - We report that the expression of mutant G93A copper/zinc superoxide dismutase (SOD1), associated with familial amyotrophic lateral sclerosis, specifically causes a decrease in MTT reduction rate and ATP levels and an increase in both cytosolic and mitochondrial reactive oxygen species (ROS) production in human neuroblastoma SH-SY5Y cells compared to cells overexpressing wild-type SOD1 and untransfected cells. Exposure to N-acetylcysteine lowers ROS production and returns mitochondrial functional assays to control levels. No large aggregates of human SOD1 are detectable under basal growth conditions in any of the investigated cell lines. After proteasome activity inhibition, SOD1 aggregates can be detected exclusively in G93A-SOD1 cells, even though they do not per se enhance cell death compared to control cell lines. Our findings indicate that mitochondrial homeostasis is affected by mutant SOD1-generated ROS independently from the formation of aggregates and that this alteration is reversed by antioxidants. PMID- 12901836 TI - Oxidative stress induces axonal beading in cultured human brain tissue. AB - Oxidative stress has been implicated in the pathogenesis of a number of human neurodegenerative disorders of the central nervous system (CNS), including Alzheimer's disease (AD). To better understand the pathological effects of oxidative stress on CNS neurons we used a primary human brain cell culture model of hydrogen peroxide-induced oxidative stress. Neuronal and astrocytic morphology was visualised by immunofluorescence with antibodies to the neuron-specific microtubule component beta-tubulin III and against glial fibrillary acidic protein (GFAP), respectively. After exposure to 40 mM H(2)O(2) for 60-90 min, axonal swelling was observed, which developed into axonal beading after 48 h. No beading was observed in GFAP-positive astrocytes. Despite the concentration of H(2)O(2) used, neurons remained attached to the substratum and showed no signs of apoptosis. This was attributed to the neuroprotective effect of the B-27 medium supplement, which contained antioxidants. The axonal swelling and beading was consistent with a disruption of microtubules by oxidative stress and subsequent hold-up of axonal transport. PMID- 12901837 TI - PML nuclear bodies and neuronal intranuclear inclusion in polyglutamine diseases. AB - In polyglutamine diseases, accumulation in the nucleus of mutant proteins induces the formation of neuronal intranuclear inclusions (NIIs). The nucleus is compartmentalized into structural and functional domains, which are involved in NII formation. Promyelocytic leukemia protein (PML), a major component of nuclear bodies, and mSin3A, a component of the transcription co-repressor complex, were used to investigate how the intranuclear domains/sites relate to NII formation in SCA2, SCA3, SCA7, SCA17 and DRPLA brains. We demonstrate that the size of PML positive intranuclear structures was larger in pathological brains than in control ones and that these structures contained mutant proteins. PML colocalized only with small NIIs, which maintained the ring-like structure of normal nuclear bodies. Enlarged ring-like PML-positive structures, devoid of mutant proteins, were also found and might represent structures where mutant polyglutamine proteins have been successfully processed. These data suggest that NIIs originate from nuclear bodies, where mutant proteins accumulate for degradation. PMID- 12901838 TI - Presenilin-1 interacts directly with the beta-site amyloid protein precursor cleaving enzyme (BACE1). AB - A neuropathological hallmark of Alzheimer's disease is the presence of amyloid plaques. The major constituent of these plaques, occurring largely in brain areas important for memory and cognition, is the 40-42 amyloid residues (Abeta). Abeta is derived from the amyloid protein precursor after cleavage by the recently identified beta-secretase (BACE1) and the putative gamma-secretase complex containing presenilin 1 (PS1). In an attempt to develop a functional secretase enzymatic assay in yeast we demonstrate a direct binding between BACE1 and PS1. This interaction was confirmed in vivo using coimmunoprecipitation and colocalization studies in human cultured cells. Our results show that PS1 preferably binds immature BACE1, thus possibly acting as a functional regulator of BACE1 maturation and/or activity. PMID- 12901839 TI - Dendritic spine loss in the hippocampus of young PDAPP and Tg2576 mice and its prevention by the ApoE2 genotype. AB - Postmortem AD brains exhibit dendritic spine loss in the hippocampus. To determine whether this pathology may be associated with amyloid burden, the present study used the Golgi stain technique to assess age- and genotype dependent changes in dendritic spine density in CA1 hippocampus of two transgenic mouse lines that produce high levels of Abeta. Tg2576 and PDAPP mice, as well as a group of Tg2576 mice crossed with human apoE2-expressing transgenic mice, were compared to respective transgene-negative controls. Since the time course of amyloid plaque deposition in the PDAPP and Tg2576 mice is well characterized, we examined changes in spine density at ages that corresponded to different levels of amyloid plaque load. The data show age- and genotype-dependent reductions in spine density in both Tg2576 and PDAPP mice, albeit at somewhat different time courses. The spine loss occurred prior to plaque deposition and was ameliorated by the overexpression of human apoE2. These results suggest that a soluble Abeta species may affect hippocampal synapses and thereby contribute to functional deficits evident in these animals. PMID- 12901840 TI - Ca2+/calmodulin-dependent modulation of cell cycle elements pRb and p27kip1 involved in the enhanced proliferation of lymphoblasts from patients with Alzheimer dementia. AB - Failure of cell cycle regulation in neurons might be critically involved in the process of neurodegeneration in Alzheimer's disease (AD). We present here evidence to support the hypothesis that cell cycle alterations occur in cells other than neurons in AD sufferers. Lymphocytes from AD patients immortalized with Epstein-Barr virus showed an enhanced rate of proliferation and increased phosphorylation of the retinoblastoma protein (pRb) and other members of the family of pocket proteins compared with cell lines derived from normal age matched controls. The calmodulin antagonist calmidazolium, as well as W-7 and W 13, abrogated the enhanced activity of AD cells without altering the normal basal rate of proliferation. The effect of calmidazolium was accompanied by partially dephosphorylation of pRb. No changes were found in the expression levels of the G1 cyclin/Cdks complexes. However, lymphoblasts derived from AD patients showed reduced levels of the Cdk inhibitor p27(kip1), which were restored after anti calmodulin treatment of the cultures. These observations suggest that in AD cells the enhanced rates of cell proliferation and phosphorylation of pRb and the intracellular content of p27(kip1) may be interrelated events controlled by a mechanism dependent on the Ca(2+)/calmodulin signaling pathway. The distinct functional features of lymphoblastoid cells from AD patients offer an invaluable, noninvasive tool to investigate the etiopathogenesis, and eventually, for the early diagnosis and prognosis of this devastating disease. PMID- 12901841 TI - Skeletal muscle properties in a transgenic mouse model for amyotrophic lateral sclerosis: effects of creatine treatment. AB - The present study was undertaken to identify the metabolic and contractile characteristics of fast- and slow-twitch skeletal muscles in a transgenic mouse model of amyotrophic lateral sclerosis (ALS). In addition, we investigated the effects of oral creatine supplementation on muscle functional capacity in this model. Transgenic mice expressing a mutant (G93A) or wild type human SOD1 gene (WT) were supplemented with 2% creatine monohydrate from 60 to 120 days of age. Body weight, rotorod performance and grip strength were evaluated. In vitro contractility was evaluated on isolated m. soleus and m. extensor digitorum longus (EDL), and muscle metabolites were determined. Body weight, rotorod performance and grip strength were markedly decreased in G93A compared to WT mice, but were unaffected by creatine supplementation. Muscle ATP content decreased and glycogen content increased in G93A versus WT in both muscle types, but were unaffected by creatine supplementation. Muscle creatine content increased following creatine intake in G93A soleus. Twitch and tetanic contractions showed markedly slower contraction and relaxation times in G93A versus WT in both muscle types, with no positive effect of creatine supplementation. EDL but not soleus of G93A mice showed significant atrophy, which was partly abolished by creatine supplementation. It is concluded that overexpression of a mutant SOD1 transgene has profound effects on metabolic and contractile properties of both fast- and slow-twitch skeletal muscles. Furthermore, creatine intake does not exert a beneficial effect on muscle function in a transgenic mouse model of ALS. PMID- 12901842 TI - ApoE4 impairs hippocampal plasticity isoform-specifically and blocks the environmental stimulation of synaptogenesis and memory. AB - Alzheimer's disease (AD) is associated with genetic risk factors, of which the allele E4 of apolipoprotein E (apoE4) is the most prevalent, and is affected by environmental factors that include education early in life and socioeconomic background. The extent to which environmental factors affect the phenotypic expression of the AD genetic risk factors is not known. Here we show that the neuronal and cognitive stimulations, which are elicited by environmental enrichment at a young age, are markedly affected by the apoE genotype. Accordingly, exposure to an enriched environment of young mice transgenic for human apoE3, which is the benign AD apoE allele, resulted in improved learning and memory, whereas mice transgenic for human apoE4 were unaffected by the enriched environment and their learning and memory were similar to those of the nonenriched apoE3 transgenic mice. These cognitive effects were associated with higher hippocampal levels of the presynaptic protein synaptophysin and of NGF in apoE3 but not apoE4 transgenic mice. In contrast, cortical synaptophysin and NGF levels of the apoE3 and apoE4 transgenic mice were similarly elevated by environmental enrichment. These findings show that apoE4 impairs hippocampal plasticity and isoform-specifically blocks the environmental stimulation of synaptogenesis and memory. This provides a novel mechanism by which environmental factors can modulate the function and phenotypic expression of the apoE genotype. PMID- 12901843 TI - Internalization of Mycobacterium tuberculosis by macropinocytosis in non phagocytic cells. AB - Mycobacterium tuberculosis (MTB) is an intracellular pathogen that initially invades the alveolar macrophages of infected individuals. MTB is also known to invade respiratory epithelial cells. To understand the mechanism of epithelial invasion, we investigated the interaction of MTB (H37Rv strain) with non phagocytic type-II (A549) human pneumocytes. The internalization of the organism was analyzed through optical, fluorescent and electron (transmission and scanning) microscopy. Infection of A549 cells with MTB showed intracellular multiplication of the organism. Microscopy revealed the formation of membrane ruffles totally or partially surrounding the surface adherent mycobacteria. Fluorescent microscopy showed that MTB induced changes in the distribution of actin filaments. Since heat killed MTB failed to induce actin mobilization, perhaps, internalization process is mediated by the soluble products of the metabolically active mycobacterium. Overall, these findings suggest that internalization of MTB by non-phagocytic cells might be through a macropinocytosis or induced-phagocytosis processes, and possibly some bacterial secretory product is responsible for triggering this phenomenon. PMID- 12901844 TI - Altering tissue tropism of Listeria monocytogenes by ectopically expressing human E-cadherin in transgenic mice. AB - The Gram-positive bacterium Listeria monocytogenes invades a variety of cells in vitro and in vivo. Here, we show that L. monocytogenes is able to enter muscle cells if provided with the appropriate host receptor. Using a cardiac-specific promoter, human E-cadherin was misexpressed in the heart of transgenic mice. First, we demonstrate that L. monocytogenes is capable of infecting the E cadherin-expressing, but not the N-cadherin-expressing cardiomyocytes in vitro. Second, we show that L. monocytogenes infects the heart of transgenic animals after administering the bacteria intravenously. This unique system may prove useful for understanding basic aspects of L. monocytogenes cell biology by comparing infection of muscle versus epithelial cells. PMID- 12901846 TI - Specific antibodies contribute to the host protection against strains of Francisella tularensis subspecies holarctica. AB - T cells are crucial to the control and eradication of the facultative intracellular bacterium Francisella tularensis. A contributory role of humoral antibodies in the host defence remains to be assessed. We used B-cell-deficient mice to study the possible contribution of antibodies to the defence against the live vaccine strain (LVS) or a clinical isolate of F. tularensis, both belonging to the subspecies holarctica (type B). When B-cell-deficient (Igmu(-/-)) mice of the C57BL/10 background were administered immune serum one day before intradermal injection of LVS, they developed lower bacterial numbers in skin, liver, and spleen than did mice receiving normal serum, and survived a challenge inoculum that was lethal for mice given normal serum. Administration of immune serum to C57BL/10 mice afforded protection also against infection with the clinical isolate of F. tularensis subsp. holarctica. Five days after intradermal inoculation of bacteria of the isolate, animals receiving immune serum showed 4log10 lower bacterial counts in liver and spleen than mice administered normal serum. In mice primed by LVS infection, T-cell immunity and host protection were strong and only a marginal contribution of immune serum against a secondary intradermal infection was demonstrated. Together, these findings show that specific antibodies contribute to the host defence of mice against F. tularensis subsp. holarctica. PMID- 12901845 TI - Influence of group A streptococcal acid glycoprotein on expression of major virulence factors and internalization by epithelial cells. AB - A single transposon insertion upstream to the open-reading-frame identified as the streptococcal acid glycoprotein (sagp) gene rendered a Tn916 isolate of Streptococcus pyogenes with elevated susceptibility to internalization by the epithelial cells. The role of SAGP in S. pyogenes internalization was further studied using isogenic mutant containing an in-frame deletion within the sagp gene. The sagp mutant displayed slower growth-rate and showed 5-fold higher internalization efficiency than the parent strain. Transcription of sagp at the logarithmic phase, but not at the stationary phase of the growth was repressed by csrR, the global regulator gene. At the same time, mutation of the sagp gene partially decreased the transcription of hasA, a gene that is required for capsule synthesis. The mutation had no effect on transcription of the emm3 gene, encoding for the M protein. The most striking effect of the sagp mutation was a down-regulation of the streptococcal pyrogenic exotoxin B (SpeB) at both translational and transcriptional level. Treatment of the SAGP mutant cells with the exogenous mSpeB (mature protease) only partially reduced their susceptibility to internalization. The exogenous mSpeB was more effective in reducing the internalization efficiency of a speB mutant and brought it to the level observed for the parent strain. In overall, results show that CsrR, directly or indirectly, affects the expression of SAGP, and that the SAGP modulates expression of not only SpeB, but also other genes that facilitate S. pyogenes internalization. PMID- 12901848 TI - Shiga toxin induces decreased expression of the anti-apoptotic protein Mcl-1 concomitant with the onset of endothelial apoptosis. AB - Shiga toxin (Stx) has been implicated in the pathogenesis of several human and animal disease states. A key host target of Stx is the endothelial cell. Stx induces endothelial cell apoptosis through a mechanism that remains unknown. In the present report, we demonstrate that Stx-1 and Stx-2 inhibit endothelial cell expression of the anti-apoptotic Bcl-2 family member, Mcl-1. Decreased expression of Mcl-1 preceded the onset of Stx-induced apoptosis. Further, Stx-1-induced decrements in Mcl-1 expression correlated in a dose-dependent manner with sensitization to Stx-1-induced apoptosis. Finally, inhibition of Mcl-1 degradation with the proteasome inhibitor, lactacystin, protected against Stx-1 induced apoptosis. These combined data suggest a role for Mcl-1 in protecting endothelial cells against Stx-1-induced apoptosis. PMID- 12901847 TI - Sodium pentobarbital anesthesia transiently enhances the severity of infection following intragastric, but not intravenous, inoculation of Listeria monocytogenes in mice. AB - In the present study we observed that mice anesthetized with sodium pentobarbital were far more susceptible to gastrointestinal challenge with Listeria monocytogenes than were unanaesthetized mice. The effect of pentobarbital anesthesia was transient (gone within 2 h) and did not alter the severity of infection following i.v. challenge with L. monocytogenes. Treatment with pharmacological inhibitors of gastric acidity (i.e. cimetidine and omeprazole), or intestinal motility (loperamide), did not duplicate the effect of pentobarbital on gastrointestinal listeriosis. These findings suggest that sodium pentobarbital anesthesia causes a short-lived but striking diminution in resistance to gastrointestinal listeriosis in mice, via an undefined mechanism. PMID- 12901849 TI - SP500263, a novel SERM, blocks osteoclastogenesis in a human bone cell model: role of IL-6 and GM-CSF. AB - Bone metabolism requires tightly coupled activities exhibited by two unique cell populations, the bone-resorbing osteoclasts and the bone-forming osteoblasts. Imbalance in the function of these two cell types can result in osteoporosis, a condition characterized by loss in bone integrity and of bone mass. We developed a human bone cell culture model that allows the in vitro study of bone formation and osteoclastogenesis and employed this bone model for the screening and pharmacological analyses of protein and small molecule therapeutics. The cytokines, interleukin-6 (IL-6) and granulocyte macrophage colony stimulating factor (GM-CSF), play an intricate role in osteoclastogenesis in this system. Neutralizing antibodies to IL-6 and GM-CSF decreased the formation of osteoclast like cells. SP500263, an early lead compound from a novel class of selective estrogen receptor modulators (SERMs), was more efficacious than estrogen and comparable to raloxifene in blocking cytokine production and formation of osteoclast-like cells. Our research demonstrates the usefulness of the in vitro co-culture model in the dissection of molecular events relevant to bone metabolism and provides greater insight into a potential novel role for cytokines in bone resorption. Furthermore, representatives of the SP500263 family of SERMs may be effective as therapeutics for the treatment of osteoporosis. PMID- 12901850 TI - Redox modulation of NO-dependent induction of interleukin 8 gene in monocytic U937 cells. AB - We have examined the effects of various antioxidants and inhibitors of redox sensitive signal transduction pathways on induction of interleukin 8 (IL-8) gene by NO in monocytic U937 cells. We have observed that nitrosoglutathione or another NO-generating compound spermine NONOate caused significant accumulation of IL-8 mRNA. Pretreatment of cells with pyrrolidine dithiocarbamate or with antioxidants, which scavenge hydroxyl radical, dimethyl sulfoxide (DMSO), or dimetylthiourea (DMTU) completely abrogated NO-dependent induction of IL-8 gene expression. The transcriptional activation of IL-8 gene was not affected by sodium formate or sodium salicylate, suggesting that suppression of the IL-8 gene induction is specific to the class of hydroxyl radical scavenger used. Furthermore, we have shown that IL-8 induction was not inhibited by catalase and the iron chelator deferoxamine, indicating that the inhibitory actions of DMSO and DMTU are not related to scavenging of reactive oxygen species produced from hydrogen peroxide in the iron-catalyzed reactions. Finally, we have not observed any significant inhibition of NO-dependent IL-8 gene induction by superoxide scavengers such as N-acetyl cysteine, uric acid, and superoxide dismutase. Therefore, it seems likely that in U937 cells, hydroxyl radicals or species with reactivity similar to hydroxyl radicals contribute to NO-mediated IL-8 gene induction. PMID- 12901851 TI - Regulation of T helper type-1 immunity in hapten-induced colitis by host pretreatment with granulocyte colony-stimulating factor. AB - Recombinant human granulocyte colony-stimulating factor (rhG-CSF) is an immunoregulatory drug whose effects include modulation of antigen-presentation. We investigated the potential ameliorative effect of pretreatment with rhG-CSF in a hapten-induced colitis animal model. Sprague-Dawley rats were given rhG-CSF (125 microg/kg subcutaneously twice a day for 5 days) before a colonic instillation of 2,4,6-trinitrobenzene sulfonic acid (TNBS) in 50% ethanol. Consequent colonic damage was evaluated pathologically, and cytokine mRNA expression levels in macroscopically inflamed sites were measured by real-time quantitative reverse transcription-polymerase chain reaction. Pretreatment with rhG-CSF remarkably attenuated both the loss of body weight and colonic wall thickening due to progressive transmural inflammation. In the control, treatment with TNBS led to a statistically significant (p < 0.05) upregulation of IFN-gamma mRNA expression in the inflammatory sites measured at post-treatment day 7. In the experimental group, pretreatment with rhG-CSF abrogated transcription of IFN gamma (p < 0.05), but was not, however, associated with an upregulation of IL-4 or the regulatory cytokines TGF-beta and IL-10. Furthermore, transcription of IL 12p35 (a rate-limiting factor for the production of IL-12) was significantly (p < 0.05) downregulated by rhG-CSF at 24h post-TNBS instillation, whereas IL-12p40 was not affected. Pretreatment with rhG-CSF drastically attenuated the degree of TNBS-induced colitis through selective downregulation of Th1-associated cytokines. PMID- 12901852 TI - Intracellular IL-1Ra type 1 inhibits IL-1-induced IL-6 and IL-8 production in Caco-2 intestinal epithelial cells through inhibition of p38 mitogen-activated protein kinase and NF-kappaB pathways. AB - Interleukin-1 (IL-1) plays a pivotal role in the pathogenesis of inflammatory bowel disease (IBD). IL-1 action is regulated in part by its naturally occurring inhibitor, the IL-1 receptor antagonist (IL-1Ra). Four splice variants of IL-1Ra gene product have been described, one secreted (sIL-1Ra) and three intracellular (icIL-1Ra1, 2, 3). Although sIL-1Ra and icIL-1Ra1 bind to type I IL-1 receptor with equal affinity, icIL-1Ra1 may carry out unique functions inside cells. The goal of this study was to determine the role of icIL-1Ra1 in regulation of cytokine-induced IL-6 and IL-8 production in Caco-2 intestinal epithelial cells. icIL-1Ra1 inhibited IL-1-induced IL-6 and IL-8 production. IL-1 activated all three mitogen-activated protein (MAP) kinase family members: p38 MAP kinase, extracellular-regulated kinases (ERK), and c-Jun amino-terminal kinases (JNK). Specific inhibitors of each MAP kinase pathway decreased IL-1-induced IL-6 and IL 8 production. Overexpression of icIL-1Ra1 inhibited p38 MAP kinase phosphorylation, but had no effect on ERK and JNK phosphorylation. In addition, icIL-1Ra1 inhibited nuclear translocation of NF-kappaB after IL-1 stimulation. In conclusion, these data indicate that icIL-1Ra1, acting in the cytoplasm of Caco-2 cells, decreased IL-1-induced IL-6 and IL-8 production. This intracellular anti inflammatory activity of icIL-1Ra1 was mediated through inhibition of p38 MAP kinase and NF-kappaB signal transduction pathways. PMID- 12901853 TI - Interleukin-1 gene cluster polymorphisms in cerebral infarction. AB - Interleukin-1 (IL-1) has pleiotropic actions in the central nervous system. During the past decade, a growing corpus of evidence has indicated an important role of this cytokine in the development of brain damage following cerebral ischaemia. The expression of IL-1 in the brain is dramatically increased during the early and chronic stage of infarction. The IL-1 gene cluster on chromosome 2ql4 contains three related genes (IL-1alpha, IL-1beta, and IL-1 receptor antagonist, IL-1ra) located within a 430-kb region. Polymorphisms in the genes encoding IL-1alpha, IL-1beta, and IL-1ra have been associated with several inflammatory diseases. Therefore we hypothesized that these cytokines might be good candidates for cerebral infarction (CI). We ascertained these genotypes in 363 CI patients and 640 controls matched for age and gender. A significant increase was found for the IL-1alpha (-889) allele 2 carriers in CI patients compared with controls (chi2 = 5.633, P = 0.018, odds ratio (OR) = 1.5). Furthermore, the IL-1alpha (-889) allele 2 carriers increased the relative risk for CI in the subjects without the IL-1ra allele 2 (chi2 = 7.989, P = 0.005, OR = 1.7). There was no significant association between IL-1beta (+3,953) polymorphism and CI. These results suggest that IL-1alpha-889 and IL-1ra polymorphisms are effective in the development of CI in Koreans. PMID- 12901854 TI - Effect of pericardial fluid pro-inflammatory cytokines on hemodynamic parameters. AB - We investigated the effects of pro-inflammatory cytokines of pericardial fluid on hemodynamic parameters in patients undergoing coronary artery surgery. Seventy eight patients were included in the study and they were allocated to three groups: group 1, stable angina pectoris (SAP, n = 15); group 2, unstable angina pectoris (USAP, n = 34); group 3, post-myocardial infarction (PMI, n = 29). Pericardial fluid and arterial blood samples were obtained from all patients and interleukin (IL)-1beta, IL-2 receptor, IL-6, IL-8 and tumor necrosis factor-alpha (TNF-alpha) levels were measured. Pericardial IL-1beta concentration (pg/mL) was significantly higher in the USAP group (26.6 +/- 10.9) compared to the SAP (5.0 +/- 0.1) and PMI (5.8 +/- 1.0) groups. IL-2R, IL-6, IL-8 and TNF-alpha concentrations of pericardial fluid were significantly higher than serum in all groups; difference was more prominent in the PMI group compared to the SAP and the USAP groups. Serum IL-1beta concentrations (pg/mL) were significantly higher in the USAP group (21.8 +/- 3.4) compared to the SAP group (5.0 +/- 0.1) and the PMI group (5.4 +/- 1.6). Cardiac index (CI) before opening the pericardial sac was found to be lower in the USAP group (1.6 +/- 0.3 L/min/m2) compared to the SAP (2.2 +/- 0.5 L/min/m2) and the PMI (2.1 +/- 0.5 L/min/m2) groups (p = 0.028 and p = 0.011, respectively). In the USAP group, there was a relationship between reduction of CI and increase of IL-1beta levels in serum and pericardial fluid. PMID- 12901855 TI - Glucose-dependent insulinotropic polypeptide analogues and their therapeutic potential for the treatment of obesity-diabetes. AB - Glucose-dependent insulinotropic polypeptide (GIP) is a key incretin hormone, released postprandially into the circulation in response to feeding, producing a glucose-dependent stimulation of insulin secretion. It is this glucose-dependency that has attracted attention towards GIP as a potential therapeutic agent for the treatment of type 2 diabetes. A major drawback to achieving this goal has been the rapid degradation of circulating GIP by the ubiquitous enzyme, dipeptidylpeptidase IV (DPP IV). However, recent studies have described a number of novel structurally modified analogues of GIP with enhanced plasma stability, insulinotropic and antihyperglycaemic activity. The purpose of this article was to provide an overview of the biological effects of several GIP modifications and to highlight the potential of such analogues in the treatment of type 2 diabetes and obesity. PMID- 12901856 TI - Selection of aptamers against KMP-11 using colloidal gold during the SELEX process. AB - SELEX procedure is a methodology in which single stranded oligonucleotides are selected from a wide variety of sequences based on their interaction with a target molecule. We have designed a novel SELEX methodology using colloidal gold to select high affinity single stranded DNA aptamers against Leishmania infantum KMP-11. Kinetoplastid membrane protein-11 (KMP-11) is a major component of the cell membrane of kinetoplastid parasites. Although its function is not known, the fact that KMP-11 is a cytoskeleton-associated protein suggests that it may be involved in mobility or in some other aspects of the flagellar structure. We have isolated a single stranded DNA aptamer population that binds specifically to L. infantum KMP-11. This population has been characterized in a series of in vitro experiments suggesting that it may be used as a powerful tool to further investigate the role of KMP-11 during Leishmania development and/or as a diagnostic tool in Leishmania infection. PMID- 12901857 TI - Structural basis of the lisinopril-binding specificity in N- and C-domains of human somatic ACE. AB - Angiotensin I-converting enzyme (ACE) is a dipeptidyl carboxypeptidase which converts angiotensin I into the vasopressor peptide angiotensin II and also inactivates the hypotensive peptide bradykinin, playing an important role in blood pressure regulation. The present work describes the molecular modeling of the N-terminal human somatic ACE in complex with the inhibitor lisinopril, identifying the residues involved in the inhibitor-binding pocket. The obtained results identify differences in the lisinopril lysine moiety-binding residues for N- and C-terminals of sACE domains and an important carboxy-terminal proline hydrophobic accommodations mediated by the aromatic ring of Tyr532 and Tyr1128 residues, respectively. The present model will be useful for the development of a new inhibitor family based on the natural BPP peptides and derivatives, or even to improve the binding capacities and the domain specificity of the already known inhibitors. PMID- 12901858 TI - Nemo-like kinase induces apoptosis in DLD-1 human colon cancer cells. AB - Deregulation of Wnt/beta-catenin signaling is thought to play a critical role in human carcinogenesis. Nemo-like kinase (NLK) is an evolutionarily conserved serine/threonine kinase that suppresses beta-catenin/T-cell factor (TCF) complex transcriptional activity through phosphorylation of TCF. Since NLK may be a tumor suppressor as a negative regulator of Wnt/beta-catenin pathway, we established tetracycline-inducible NLK and its kinase-negative mutant expression in DLD-1 human colon cancer cells to analyze the effect of NLK on cell growth and viability. The induction of wild-type NLK in DLD-1 cells caused suppression of cell growth whereas the kinase-negative mutant did not. Flow cytometry indicated that NLK expression increased the number of apoptotic cells but did not induce obvious cell cycle arrest. Apoptosis induction by wild-type NLK was confirmed using TUNEL assays. Our results suggest that overexpression of NLK may have targets other than TCF for induction of apoptosis in human colon carcinoma cells. PMID- 12901859 TI - Characterisation of CadR from Pseudomonas aeruginosa: a Cd(II)-responsive MerR homologue. AB - cadR from Pseudomonas aeruginosa encodes a transcriptional regulatory protein which responds to Cd(II)>>Zn(II)>Hg(II) at its cognate promoter PcadA. CadR will also act to induce transcription at the Escherichia coli ZntR cognate promoter, PzntA, however, the induction profile is altered to Hg(II)>Cd(II)>Zn(II). Two separate single base pair deletions within PzntA result in further alteration of relative specificity in metal-ion induction profile for CadR. This demonstrates that the operator/promoter sequence can play a role in defining optimal ligand response and that for these regulators specificity is not solely a function of the regulatory protein. PMID- 12901860 TI - A real-time immuno-PCR assay for routine ultrasensitive quantification of proteins. AB - A fast and robust assay, based on the combination of the highly sensitive immuno PCR (IPCR), employing standardized self-assembled DNA-protein conjugates as reagents, and the well-established, reliable, and fast real-time PCR detection by means of the TaqMan principle is introduced in this work. The use of anti-species immunoglobulin reagents allows one for easy adaptation of this assay to basically any existing ELISA application. The use of an internal competitor in the real time IPCR (rtIPCR) further increases the sensitivity and significance of this assay; 0.1-0.01 amol (500-50 fg/mL) IgG from several species (mouse, rabbit, goat, and human) were detectable using direct, indirect, and sandwich model rtIPCR assays, thereby increasing the detection limit of the analogous ELISA tests about 100- to 1000-fold. The robustness of this method was demonstrated in two typical applications by detecting 40 pg/mL of the novel anti-cancer drug rViscumin in human plasma samples as well as 100 pg/mL of a research antibody in cell culture media. In both cases, a comparable ELISA was 1000-fold less sensitive. PMID- 12901861 TI - Downregulation and growth inhibitory effect of epithelial-type Kruppel-like transcription factor KLF4, but not KLF5, in bladder cancer. AB - Kruppel-like factors (KLFs) are key transcriptional regulators of cell differentiation and proliferation. Among the KLF family, the expression of KLF4 (GKLF) and KLF5 (IKLF) is highly restricted in the epithelial cells of several organs such as the gut and skin, and it has been reported that these epithelial type KLF genes may be involved in colon carcinogenesis. Recently we found that Klf4 and Klf5 genes were significantly expressed in the developmental bladder epithelium of mice as well. Therefore, in this report we studied the involvement of the KLF4 and KLF5 genes in bladder carcinogenesis. First, we analyzed the expression of KLF4 and KLF5 in a variety of human bladder cancer cell lines and surgical specimens by RNA blot and in situ hybridization analyses. Both genes were highly expressed in the normal bladder epithelium, whereas KLF4, but not KLF5, was frequently downregulated in bladder cancer cell lines and cancer tissues. We then transduced the KLF4 and KLF5 genes into the bladder cancer cell lines using adenoviral vectors to examine the biological activities of the genes on those cells. The transduction of KLF4, but not KLF5, suppressed cell growth and induced apoptosis. Our study suggests that inactivation of KLF4 is one of the frequent steps towards bladder carcinogenesis. PMID- 12901862 TI - Inhibition of 11 beta-hydroxysteroid dehydrogenase type 2 by dithiocarbamates. AB - Dithiocarbamates (DTCs), important therapeutic and industrial chemicals released in high quantities into the environment, exhibit complex chemical and biological activities. Here, we demonstrate an effect of DTCs on glucocorticoid action due to inhibition of 11 beta-hydroxysteroid dehydrogenase (11 beta-HSD) type 2, converting cortisol to cortisone in the kidney, but not 11 beta-HSD1, catalyzing the reverse reaction in liver and adipose tissue. Thus, DTCs may locally increase active glucocorticoid concentrations. Preincubation with the DTC thiram abolished 11 beta-HSD2 activity, suggesting irreversible enzyme inhibition. The sulfhydryl protecting reagent dithiothreitol blocked thiram-induced inhibition and NAD+ partially protected 11 beta-HSD2 activity, indicating that DTCs act at the cofactor-binding site. A 3D-model of 11 beta-HSD2 identified Cys90 in the NAD(+) binding site as a likely target of DTCs, which was supported by a 99% reduced activity of mutant Cys90 to serine. The interference of DTCs with glucocorticoid mediated responses suggests a cautious approach in the use of DTCs in therapeutic applications and in exposure to sources of DTCs such as cosmetics and agricultural products by pregnant women and others. PMID- 12901863 TI - Subcellular localization and N-glycosylation of human ABCC6, expressed in MDCKII cells. AB - Mutations in the gene coding for a human ABC transporter protein, ABCC6 (MRP6), are responsible for the development of pseudoxanthoma elasticum. Here, we demonstrate that human ABCC6, when expressed by retroviral transduction in polarized mammalian (MDCKII) cells, is exclusively localized to the basolateral membrane. The human ABCC6 in MDCKII cells was found to be glycosylated, in contrast to the underglycosylated form of the protein, as expressed in Sf9 cells. In order to localize the major glycosylation site(s) in ABCC6, we applied limited proteolysis on the fully glycosylated and underglycosylated forms, followed by immunodetection with region-specific antibodies for ABCC6. Our results indicate that Asn15, which is located in the extracellular N-terminal region of human ABCC6, is the only N-glycosylation site in this protein. The polarized mammalian expression system characterized here provides a useful tool for further examination of routing, glycosylation, and function of the normal and pathological variants of human ABCC6. PMID- 12901865 TI - Cells lacking Pcp1p/Ugo2p, a rhomboid-like protease required for Mgm1p processing, lose mtDNA and mitochondrial structure in a Dnm1p-dependent manner, but remain competent for mitochondrial fusion. AB - The dynamin-related GTPase, Mgm1p, is critical for the fusion of the mitochondrial outer membrane, maintenance of mitochondrial DNA (mtDNA), formation of normal inner membrane structures, and inheritance of mitochondria. Although there are two forms of Mgm1p, 100 and 90 kDa, their respective functions and the mechanism by which these two forms are produced are not clear. We previously isolated ugo2 mutants in a genetic screen to identify components involved in mitochondrial fusion [J. Cell Biol. 152 (2001) 1123]. In this paper, we show that ugo2 mutants are defective in PCP1, a gene encoding a rhomboid-related serine protease. Cells lacking Pcp1p are defective in the processing of Mgm1p and produce only the larger (100 kDa) form of Mgm1p. Similar to mgm1delta cells, pcp1delta cells contain partially fragmented mitochondria, instead of the long tubular branched mitochondria of wild-type cells. In addition, pcp1delta cells, like mgm1delta cells, lack mtDNA and therefore are unable to grow on nonfermentable medium. Mutations in the catalytic domain lead to complete loss of Pcp1p function. Similar to mgm1delta cells, the fragmentation of mitochondria and loss of mtDNA of pcp1delta cells were rescued when mitochondrial division was blocked by inactivating Dnm1p, a dynamin-related GTPase. Surprisingly, in contrast to mgm1delta cells, which are completely defective in mitochondrial fusion, pcp1delta cells can fuse their mitochondria after yeast cell mating. Our study demonstrates that Pcp1p is required for the processing of Mgm1p and controls normal mitochondrial shape and mtDNA maintenance by producing the 90 kDa form of Mgm1p. However, the processing of Mgm1p is not strictly required for mitochondrial fusion, indicating that the 100 kDa form is sufficient to promote fusion. PMID- 12901864 TI - Distinct roles of Rab3B and Rab13 in the polarized transport of apical, basolateral, and tight junctional membrane proteins to the plasma membrane. AB - Regulated transport of proteins to distinct plasma membrane domains is essential for the establishment and maintenance of cell polarity in all eukaryotic cells. The Rab family small G proteins play a crucial role in determining the specificity of vesicular transport pathways. Rab3B and Rab13 localize to tight junction in polarized epithelial cells and cytoplasmic vesicular structures in non-polarized fibroblasts, but their functions are poorly understood. Here we examined their roles in regulating the cell-surface transport of apical p75 neurotrophin receptor (p75NTR), basolateral low-density lipoprotein receptor (LDLR), and tight junctional Claudin-1 using transport assay in non-polarized fibroblasts. Overexpression of Rab3B mutants inhibited the cell-surface transport of LDLR, but not p75NTR and Claudin-1. In contrast, overexpression of Rab13 mutants impaired the transport of Claudin-1, but not LDLR and p75NTR. These results suggest that Rab3B and Rab13 direct the cell-surface transport of LDLR and Claudin-1, respectively, and may contribute to epithelial polarization. PMID- 12901867 TI - Toxicity of melanin-free ink of Sepia officinalis to transformed cell lines: identification of the active factor as tyrosinase. AB - The melanin-free ink of the cephalopod Sepia officinalis is shown to contain a heat labile proteinaceous component toxic to a variety of cell lines, including PC12 cells. Gel filtration chromatography indicated that the toxic component was concentrated in those fractions eluted at a molecular weight higher than 100 kDa and exhibiting the highest tyrosinase activity. SDS-PAGE analysis of the active fractions displayed a single major band migrating at an approximate molecular weight of 100 kDa, identical with that of the single tyrosinase band in the melanin-free ink. These data unambiguously demonstrated the identity of the toxic component with tyrosinase. Treatment of purified Sepia as well as of mushroom tyrosinase with an immobilized version of proteinase K resulted in a parallel loss of tyrosinase activity and cytotoxicity. Sepia apotyrosinase was ineffective in inducing cytotoxicity in PC12 cells. Purified Sepia tyrosinase was found to induce a significant increase in caspase 3 activity in PC12 cells, leading eventually to an irreversible apoptotic process. Overall, these results disclose a hitherto unrecognized property of tyrosinase that may lead to a reappraisal of its biological significance beyond that of a mere pigment producing enzyme. PMID- 12901866 TI - Norepinephrine induces hepatic fibrogenesis in leptin deficient ob/ob mice. AB - Leptin's actions on certain cells require a leptin-inducible neurotransmitter, norepinephrine (NE). NE modulates hepatic fibrosis. Therefore, decreased NE may explain why leptin deficiency inhibits hepatic fibrosis. We manipulated adrenergic activity in leptin-deficient ob/ob mice, leptin-sufficient, dopamine beta-hydroxylase deficient (Dbh(-/-)) mice, and HSC cultures to determine if leptin requires NE to activate HSC and induce hepatic fibrosis. ob/ob mice have chronic liver injury, but reduced numbers of HSC. Supplemental leptin increases HSC, suggesting that leptin-dependent, injury-related factors permit expansion of HSC populations. NE also increases HSC numbers and activation, normalizing fibrogenesis. When fed hepatotoxic diets, NE-deficient Dbh(-/-) mice fail to accumulate activated HSC and have impaired fibrogenesis unless treated with adrenergic agonists. NE acts directly on HSC to modulate leptin's actions because leptin increases HSC proliferation and prazosin, an alpha-adrenoceptor antagonist, inhibits this. Thus, leptin permits injury-related increases in adrenergic activity and requires NE to activate HSC and induce hepatic fibrogenesis. PMID- 12901868 TI - The conserved P1' Ser of Bowman-Birk-type proteinase inhibitors is not essential for the integrity of the reactive site loop. AB - The isolated reactive site beta-hairpin loop of Bowman-Birk-type proteinase inhibitors has become a widely studied proteinomimetic because it retains the three-dimensional structure and much of the inhibitory potency of the corresponding region of the complete protein. Here we analyse the role of the P1' Ser residue which is highly conserved and intramolecularly hydrogen bonded in the complete proteins. A combined kinetic and structural analysis of variant proteinomimetic peptides demonstrates that the hydrogen-bond potential of the side-chain oxygen atom of the P1' Ser is not essential for the integrity of the reactive site loop and that it provides only a small contribution to the trypsin affinity and no apparent contribution to the stability against tryptic turnover. We conclude that the potential of the P1' side chain to engineer improved inhibition and selectivity for serine proteinases is best explored further in concert with the side chains of the P2 and P5' residues which may interact or compete for the same space. PMID- 12901869 TI - Stretch-induced IL-6 secretion from endothelial cells requires NF-kappaB activation. AB - Interleukin-6 (IL-6) secretion from endothelial cells (ECs) in response to mechanical stimuli plays an important role in the regenerative and inflammatory responses. The aim of this study was to determine the mechanism for the secretion of IL-6 from ECs in response to uni-axial continuous stretch. Continuous stretch induced IL-6 secretion from human umbilical vein endothelial cells (HUVECs) in a dose-dependent manner. Reverse transcriptase-polymerase chain reaction (RT-PCR) amplification showed that the transcription of the IL-6 gene peaked 2h after stretch. In vitro kinase assay of IkappaB kinase (IKKs) activity demonstrated that the activation of IKKs peaked 15 min after stretch. Two NF-kappaB inhibitors, pyrrolidine dithiocarbamanate (PDTC) and SN50, or antisense oligodeoxynucleotides for NF-kappaB p65 and p50 suppressed IL-6 mRNA expressions induced by continuous stretch. In conclusion, continuous stretch induces IL-6 secretion from ECs, most likely through sequential activation of IKKs and NF kappaB. PMID- 12901870 TI - Myelin basic protein has multiple calmodulin-binding sites. AB - Myelin basic protein (MBP) has been shown to bind calmodulin (CaM) in a specific Ca(2+)-dependent manner via a primary target sequence at its C-terminus [Protein Sci. 12 (2003) 1507]. Upon deimination of MBP, the nature of the interaction changed significantly, suggesting either a new binding site or different conformers with different affinities for CaM. In order to resolve this issue, we investigated here the CaM-binding properties of N- and C-terminal deletion mutants of MBP using Trp fluorescence spectroscopy and mass spectrometry. We conclude that there is an additional CaM-binding site on MBP in a central segment (we posit murine residues 82-93) that forms an amphipathic alpha-helix. PMID- 12901871 TI - Tartrate-resistant acid phosphatase 5B circulates in human serum in complex with alpha2-macroglobulin and calcium. AB - Tartrate-resistant acid phosphatase (TRACP) is an enzyme with unknown biological function. In addition to its acid phosphatase activity, TRACP is capable of generating reactive oxygen species (ROS) at neutral pH. Two forms of TRACP circulate in human serum, macrophage-derived TRACP 5a and osteoclast-derived TRACP 5b. Here we have studied the circulating forms of the osteoclast-derived TRACP 5b in rat and human serum. In human serum, TRACP 5b circulates in a large complex that contained alpha2M and calcium. On the contrary, rat serum TRACP 5b circulates as a free molecule. Formation of the TRACP 5b complex in vitro decreased significantly the ROS generating activity of TRACP 5b without affecting its phosphatase activity. These results suggest that the complex formation may be necessary to eliminate the formation of the harmful ROS in the neutral pH of serum. PMID- 12901872 TI - Role of Stat5 in type I interferon-signaling and transcriptional regulation. AB - Type I interferons are pleiotropic cytokines that transduce signals via activation of multiple downstream signaling cascades, including the Jak-Stat pathway. Although the roles of Stat1 and Stat2 in Type I interferon signaling are well established, the roles that other Stat-family members play in the induction of IFN-responses remain to be defined. In previous studies, we have shown that Stat5 associates with the CrkL adapter and forms a signaling complex that binds DNA. In the present study, we provide evidence that Stat5 is phosphorylated on serines 725/730 in an IFNalpha- and IFNbeta-dependent manner, providing direct evidence that serine phosphorylation of the protein is a component of an interferon signaling cascade. Such serine phosphorylation of Stat5 is Map kinase- and PI 3(')-kinase independent, while the activation of the serine kinase that phosphorylates Stat5 is regulated by upstream tyrosine kinase activity. Using mouse embryonic fibroblasts with targeted disruption of the Stat5a and Stat5b genes, we demonstrate that full activation of Stat5 is required for Type I interferon-dependent gene transcription via GAS elements. Altogether, our data provide evidence that Stat5 plays an important role in IFN-signaling and participates in the induction of Type I IFN-dependent responses. Furthermore, our results strongly suggest that, in addition to phosphorylation on tyrosine residues, phosphorylation on serine residues exhibits regulatory effects on the transcriptional capacity of Stat5. PMID- 12901873 TI - TACE/ADAM-17 enzymatic activity is increased in response to cellular stimulation. AB - Tumor necrosis factor-alpha converting enzyme (TACE/ADAM-17) is a metalloprotease disintegrin that cleaves a variety of membrane proteins, releasing ("shedding") their extracellular domains from cells. Most TACE-mediated shedding events occur at low basal rates that are enhanced by treatment of cells with a variety of stimuli. To study the mechanism of induced shedding, we developed a peptide cleavage assay that measures the cellular TACE activity. In unstimulated cells, cleavage of a TNFalpha processing-site peptide was mediated mainly by enzymes other than TACE. However, stimulation of cells with phorbol-12-myristate-13 acetate (PMA) increased peptide cleavage in a TACE-dependent manner. PMA treatment did not increase the amount of TACE on the cell surface. Moreover, the cytoplasmic domain of TACE was not required for the induced activity. Based on these observations, induction of TACE-mediated shedding events occurs at least in part via an increase in the enzymatic activity of cellular TACE, independent of its cytoplasmic domain. PMID- 12901874 TI - Vitamin C supplementation in normal subjects reduces constitutive ICAM-1 expression. AB - Regulation of monocyte adhesion molecule gene expression is via redox sensitive transcription factors. We have investigated whether dietary antioxidant supplementation with vitamin C (250 mg/day) can modulate monocyte ICAM-1 expression in healthy male subjects with low plasma vitamin C at baseline. In a randomised, double-blind, crossover study, monocyte ICAM-1 mRNA was analysed using quantitative reverse transcriptase PCR. Protein was determined by flow cytometry (monocytes) and ELISA (plasma). Monocyte numbers were unaltered by supplementation. Subjects with low plasma vitamin C (<50 microM) prior to supplementation expressed higher levels of monocyte ICAM-1mRNA, and showed a significant (50%) reduction in ICAM-1mRNA expression after 6 weeks of 250 mg/day vitamin C supplementation (p<0.05). This was paralleled by a reduction in sICAM-1 (p<0.05). For the first time, these results show that dietary vitamin C can modulate monocyte ICAM-1 gene expression in vivo, where regulation of gene expression represents a novel mechanism for benefit from dietary antioxidants. PMID- 12901875 TI - Proteasome inhibition by chronic oxidative stress in human trabecular meshwork cells. AB - The pathophysiologic mechanisms leading to the malfunction of the trabecular meshwork (TM)-Schlemm's canal (SC) outflow pathway in glaucoma are still unclear. We hypothesize that chronic oxidative stress may contribute to the malfunction of the outflow pathway by impairing the intracellular proteasome system of the cells, decreasing the ability of the tissue to modulate outflow resistance. To study the effects of chronic oxidative stress on proteasome function, primary cultures of human TM cells were incubated under 40% oxygen and proteasome activity was analyzed by measuring the accumulation of enhanced green fluorescent protein fused to a PEST motif. Changes in proteasome content, cellular senescence, and cell viability were also monitored. After 10 days of exposure to chronic oxidative stress, TM cells showed a marked decline in proteasome activity that was associated with premature senescence and decreased cell viability. These results suggest that proteasome failure may be involved in glaucoma pathophysiology. PMID- 12901876 TI - Raf1 plays a pivotal role in lipopolysaccharide-induced activation of dendritic cells. AB - Activation of extracellular-regulated kinases 1/2 (ERK) is involved in lipopolysaccharide (LPS)-induced cellular responses such as the increased production of proinflammatory cytokines. However, mitogen-activated protein kinases (MAPKs) such as p38 are also activated by LPS and have been postulated to be important in the control of these end points. Therefore, establishing the relative contribution of MAPKs in each cell type is important, as is elucidating the molecular mechanisms by which these MAPKs are activated in LPS-induced signaling cascades. We demonstrated in DC2.4 dendritic cells that ERK regulates tyrosine phosphorylation of phosphatidyl-inositol-3-kinase (PI3-K) and the production of TNF-alpha. We also demonstrated that Raf1 is phosphorylated and involved in the production of TNF-alpha and tyrosine phosphorylation of PI3-K via ERK. Raf1 also regulates the activation of NF-kappaB. We propose that Raf1 plays a pivotal role in LPS-induced activation of the dendritic cells. PMID- 12901877 TI - PPARdelta activation induces COX-2 gene expression and cell proliferation in human hepatocellular carcinoma cells. AB - Cyclooxygenase-2 (COX-2) has been suggested to be associated with carcinogenesis. Recently, many studies have shown increased expression of COX-2 in a variety of human malignancies, including hepatocellular carcinoma (HCC). Therefore, it becomes important to know more about what determines COX-2 expression. In this work, we have studied the effect of PPARdelta activation on COX-2 expression using a selective agonist (GW501516) in human hepatocellular carcinoma (HepG2) cells. Activation of PPARdelta resulted in increased COX-2 mRNA and protein expression. The mechanism behind the induction seems to be increased activity of the proximal promoter of the COX-2 gene, spanning nucleotides -327 to +59. The increased COX-2 protein expression and promoter activity induced by the GW501516 was also confirmed in the monocytic cell line THP-1. Induced levels of COX-2 have previously been associated with resistance to apoptosis and increased cell proliferation in many cell types. In HepG2 cells, we observed a dose-dependent increase in cell number by GW501516 treatment for 72h. The levels of PCNA, used as an indicator of cell division were induced, and the cell survival promoting complex p65 (NF-kappaB) was phosphorylated under GW501516 treatment. We conclude that PPARdelta activation in HepG2 cells results in induced COX-2 expression and increased cellular proliferation. These results may suggest that PPARdelta plays an important role in the development of HCC by modulating expression of COX-2. PMID- 12901878 TI - Cystatin-B is expressed by neural stem cells and by differentiated neurons and astrocytes. AB - Mutation in the gene encoding cystatin-B (CSTB) has been shown to cause progressive myoclonus epilepsy. Mice with a gene deletion of CSTB exhibit increased apoptosis of specific neurons but the physiological role of CSTB in brain cells is not fully understood. In the present study, we have examined the expression of CSTB in neural stem cells (NSC) and in differentiating mature brain cells. The results show that CSTB is present in embryonic and adult NSC and in the neuroepithelium. CSTB was expressed by both neurons and glial cells differentiated from NSC and in hippocampal cultures. CSTB localized mainly to the nucleus in NSC and in neurons, whilst in astrocytes CSTB was also in the cytoplasm. Double labeling showed that CSTB was present in the lysosomes in glial cells. The results demonstrate a nuclear expression of CSTB in NSC and in neurons, suggesting novel function for this molecule. PMID- 12901879 TI - A simple and rapid method to determine the zygosity of uPA-transgenic SCID mice. AB - Successful transplantation of xenogeneic hepatocytes into uPA-transgenic SCID mice depends on the zygosity of the recipient mice. Normally, the difference between homozygous and heterozygous animals is determined via a quantitative Southern blot. We sequenced a part of the mouse genome that is eliminated upon integration of the transgene in the genome. Based on that sequence we developed a multiplex PCR that allows the unambiguous discrimination of negative, heterozygous, and homozygous uPA-transgenic SCID mice in a single day procedure. The speed of the procedure is an essential quality because transplantation of xenogeneic hepatocytes into uPA-SCID mice should be done as soon as possible after birth. PMID- 12901880 TI - BNIPL-2, a novel homologue of BNIP-2, interacts with Bcl-2 and Cdc42GAP in apoptosis. AB - The execution phase of apoptosis is characterized by marked changes in cell morphology that include contraction and membrane blebbing. Little is known about the mechanisms underlying this process. We report here the identification of a novel member of BNIPL family, designated Bcl-2/adenovirus E1B 19kDa interacting protein 2 like-2 (BNIPL-2), which interacts with Bcl-2 and Cdc42GAP. We found that the human BNIPL-2 shares homology to human BNIP-2 and also possesses a BNIP 2 and Cdc42GAP homology (BCH) domain. Deletion experiments indicated that the BCH domain of BNIPL-2 is critical for its interactions with the Bcl-2 and Cdc42GAP and also for its cell death-inducing function. Our data showed that BNIPL-2 may be a linker protein located at the front end of Bcl-2 pathway for DNA fragmentation and Cdc42 signaling for morphological changes during apoptosis. We propose that BNIPL-2 protein may play an important role in regulation of both pathways for DNA fragmentation and for formation of membrane blebs in apoptotic cells. PMID- 12901881 TI - Pro-MMP-9 activation by the MT1-MMP/MMP-2 axis and MMP-3: role of TIMP-2 and plasma membranes. AB - MMP-9 (gelatinase B) is produced in a latent form (pro-MMP-9) that requires activation to achieve catalytic activity. Previously, we showed that MMP-2 (gelatinase A) is an activator of pro-MMP-9 in solution. However, in cultured cells pro-MMP-9 remains in a latent form even in the presence of MMP-2. Since pro MMP-2 is activated on the cell surface by MT1-MMP in a process that requires TIMP 2, we investigated the role of the MT1-MMP/MMP-2 axis and TIMPs in mediating pro MMP-9 activation. Full pro-MMP-9 activation was accomplished via a cascade of zymogen activation initiated by MT1-MMP and mediated by MMP-2 in a process that is tightly regulated by TIMPs. We show that TIMP-2 by regulating pro-MMP-2 activation can also act as a positive regulator of pro-MMP-9 activation. Also, activation of pro-MMP-9 by MMP-2 or MMP-3 was more efficient in the presence of purified plasma membrane fractions than activation in a soluble phase or in live cells, suggesting that concentration of pro-MMP-9 in the pericellular space may favor activation and catalytic competence. PMID- 12901882 TI - Structural and functional characterization of the GalNAc/Gal-specific lectin from the phytopathogenic ascomycete Sclerotinia sclerotiorum (Lib.) de Bary. AB - The lectin found in mycelium and sclerotes of the phytopathogenic fungus Sclerotinia sclerotiorum is a homodimer consisting of two identical non covalently bound subunits of 16,000 Da. CD spectra analysis revealed that the S. sclerotiorum agglutinin (SSA) contains predominantly beta-sheet structures. SSA exhibits specificity towards GalNAc whereby the hydroxyls at positions 4 and 6 of the pyranose ring play a key role in the interaction with simple sugars. The carbohydrate-binding site of SSA can also accommodate disaccharides. The N terminal sequence of SSA shares no significant similarity with any other protein except a lectin from the Sclerotiniaceae species Ciborinia camelliae. A comparison of SSA and the lectins from C. camelliae and some previously characterized lectins indicates that the Sclerotiniaceae lectins form a homogeneous family of fungal lectins. This newly identified lectin family, which is structurally unrelated to any other family of fungal lectins, is most probably confined to the Ascomycota. PMID- 12901884 TI - 20 years of hard lessons. PMID- 12901883 TI - Chitosan inhibits prostaglandin E2 formation and cyclooxygenase-2 induction in lipopolysaccharide-treated RAW 264.7 macrophages. AB - Chitosan, a deacetylated chitin, has been reported to accelerate the wound healing and exert anti-inflammatory effect but the possible mechanisms involved are still unclear. Enhanced production of prostaglandin E2 (PGE2) and pro inflammatory cytokines has been shown to contribute to immunosuppression and cytotoxicity during wound healing. In this study, we examined the effect of chitosan on cyclooxygenase pathway and cytokines production in lipopolysaccharide (LPS)-treated RAW 264.7 macrophages. Our results first demonstrated that chitosans (MW=50,000, 150,000 or 300,000) significantly inhibit the overproduction of PGE2 as well as cyclooxygenase-2 (COX-2) protein expression and activity accompanied by attenuation of pro-inflammatory cytokines production such as tumor necrosis factor-alpha and interleukin-1beta formation but increase of the anti-inflammatory cytokine, IL-10, formation in LPS-treated RAW 264.7 macrophages. These results suggest that the beneficial effect of chitosan on wound healing may be associated, at least partly, with the inhibition of PGE2 production by suppressing COX-2 induction and activity as well as attenuation of pro-inflammatory/anti-inflammatory cytokines ratio in activated macrophages. PMID- 12901885 TI - The problems of allocating large sums of money for AIDS. PMID- 12901887 TI - Scaling-up and sustaining insecticide-treated net coverage. PMID- 12901886 TI - Scaling-up and sustaining insecticide-treated net coverage. PMID- 12901888 TI - Judge rules in favour of MMR vaccine. PMID- 12901889 TI - Papillomavirus vaccines in clinical trials. AB - Cervical cancer remains a leading cause of death for women in the developing world, and the treatment of preneoplastic cervical lesions is a considerable public-health burden in the developed world. There is unambiguous evidence that human papillomaviruses (HPVs) trigger the development of cervical and other anogenital malignancies, and that continued expression of HPV antigens in the tumours drives the neoplastic progression. The viral cause of cervical cancer is also its Achilles heel. Prophylactic vaccines to prevent HPV infection and therapeutic vaccines targeted at the HPV tumour antigens are in clinical trials. A firm grasp of the molecular pathogenesis of HPVs and the natural history of genital HPV infections, combined with greater understanding of how to trigger effective immune responses, offers hope for the elimination of HPV-associated diseases. PMID- 12901890 TI - Evaluation of outcomes in community-acquired pneumonia: a guide for patients, physicians, and policy-makers. AB - Community-acquired pneumonia (CAP) is a key target for research and quality improvement in acute medicine. However, many of the outcome measures used in prognostic and antibiotic studies are not validated and do not capture features of outcome that are important to patients. Substitutes for traditional outcome measures include a recently validated patient-based symptom questionnaire (the CAP-Sym) and process-of-care measures. The interpretation of outcomes also depends on the quality of the study design and methods used. This paper discusses the advantages and disadvantages of outcome, process-of-care, and economic measures in CAP and the interpretation of these measures in randomised and observational studies. A core set of measures for use in clinical CAP research and performance measurement is proposed. PMID- 12901891 TI - Lyme borreliosis. AB - Lyme borreliosis is a multi-organ infection caused by spirochetes of the Borrelia burgdorferi sensu lato group with its species B burgdorferi sensu stricto, Borrelia garinii, and Borrelia afzelii, which are transmitted by ticks of the species Ixodes. Laboratory testing of Lyme borreliosis includes culture, antibody detection using ELISA with whole extracts or recombinant chimeric borrelia proteins, immunoblot, and PCR with different levels of sensitivity and specificity for each test. Common skin manifestations of Lyme borreliosis include erythema migrans, lymphocytoma, and acrodermatitis chronica atrophicans. The last two conditions are usually caused by B garinii and B afzelii, respectively, which are seen more frequently in Europe than in America. Late extracutaneous manifestations of Lyme borreliosis are characterised by carditis, neuroborreliosis, and arthritis. We present evidence-based treatment recommendations for Lyme borreliosis and review the prevention of Lyme borreliosis, including the Lyme vaccines. PMID- 12901892 TI - Antibacterial cleaning and hygiene products as an emerging risk factor for antibiotic resistance in the community. AB - Antibiotic resistance within the community setting is an emerging public-health concern. Infection with antibiotic-resistant organisms in the community among people lacking traditional risk factors has been reported. In addition, prevalence studies have identified individuals carrying antibiotic-resistant organisms in the absence of known risk factors. These studies strongly suggest the presence of contributing factors that have yet to be identified. In this paper we review the burden of antibiotic resistance and known risk factors within the community setting, assess the potential role of antibacterial cleaning and hygiene products containing triclosan in the emergence of resistance, and recommend future research on the assessment of household cleaning and hygiene products containing triclosan. PMID- 12901893 TI - Is Crohn's disease caused by a mycobacterium? Comparisons with leprosy, tuberculosis, and Johne's disease. AB - Although Crohn's disease is considered to be autoimmune in origin, there is increasing evidence that it may have an infectious cause. The most plausible candidate is Mycobacterium avium subspecies paratuberculosis (MAP). Intriguingly, Koch's postulates may have been fulfilled for MAP and Crohn's disease, even though they still have not been met for Mycobacterium leprae and leprosy. In animals MAP causes Johne's disease, a chronic wasting intestinal diarrhoeal disease evocative of Crohn's disease. Johne's disease occurs in wild and domesticated animals, including dairy herds. Viable MAP is found in human and cow milk, and is not reliably killed by standard pasteurisation. MAP is ubiquitous in the environment including in potable water. Since cell-wall-deficient MAP usually cannot be identified by Ziehl-Neelsen staining, identification of MAP in human beings requires culture or detection of MAP DNA or RNA. If infectious in origin, Crohn's disease should be curable with appropriate antibiotics. Many studies that argue against a causative role for MAP in Crohn's disease have used antibiotics that are inactive against MAP. However, trials that include macrolide antibiotics indicate that a cure for Crohn's disease is possible. The necessary length of therapy remains to be determined. Mycobacterial diseases have protean clinical manifestations, as does Crohn's disease. The necessity of stratifying Crohn's disease into two clinical manifestations (perforating and non-perforating) when interpreting the results of antibiotic therapy is discussed. Rational studies to evaluate appropriate therapies to cure Crohn's disease are proposed. PMID- 12901894 TI - HIV vaccine still out of our grasp. PMID- 12901895 TI - Abigail Salyers--Professor of Microbiology at the University of Illinois. Interviewed by Pam Das. PMID- 12901896 TI - Ethiopia loses HIV research centre. PMID- 12901897 TI - Malaria in Iraq---the pitfalls of Plasmodium vivax prophylaxis. PMID- 12901898 TI - Whale meat again. PMID- 12901900 TI - Carbohydrate transition state mimics: synthesis of imidazolo-pyrrolidinoses as potential nectrisine surrogates. AB - The syntheses of four glyco-imidazoles, which are pentose-derivatives belonging to the D-series, as well as the syntheses of their L-enantiomers, are reported. Starting from the known linear xylo, lyxo, arabino, and ribo imidazolo-pentoses in both the L- and the D-series, intramolecular Walden inversion led to the corresponding arabino, ribo, xylo, and lyxo pyrrolidinopentoses in the D- and the L-series, respectively, protection and deprotection steps being unavoidable prerequisites. The structures and configurations of all eight pyrrolidinopentoses were determined unambiguously, by a combination of 1H/13C NMR spectroscopy, circular dichroism and [alpha](D) values, in conjunction with single-crystal X ray diffraction analysis of the L-xylo stereoisomer. Examination of the inhibitory properties of these imidazolo-pyrrolidinoses against six commonly encountered glycosidases led to the conclusion that by and large the L stereoisomers are inactive, whereas three out the four D-stereoisomers proved to be poor to moderate inhibitors. It appears therefore that the most basic N(1) atom is not located in an optimal topology to be protonated easily inside the enzyme's active site. PMID- 12901901 TI - Physicochemical prediction of a brain-blood distribution profile in polycyclic amines. AB - Recent investigation into the pharmacological character of the pentacyclo[5.4.0.0(2,6).0(3,10).0(5,9)]undecyl and related polycyclic amines has revealed interesting facts regarding their possible use as neuroprotective agents. At this stage however, a clear shortcoming in the quest for further development of this novel class of compounds is the lack of concrete data on their ability to cross the blood-brain barrier (BBB). Working towards the aim of predicting BBB permeability, a series of related N-substituted 8-amino-8,11 oxapentacyclo[5.4.0.0(2,6).0(3,10).0(5,9)]undecanes were synthesised. Compounds were characterised by both experimental and calculative methods, followed by biological assessment and statistical manipulation of the results obtained. In doing so, a simple biological model was established for the comparative evaluation of brain-blood distribution properties within the class. A highly sensitive ESI-MS.MS analytical procedure was developed for the detection of these compounds in biological tissues, indicating significant drug concentrations in the brain after intraperitoneal administration to C57Bl/6 mice. Stepwise multiple linear regression analysis of all data yielded two meaningful models (R(2)=0.9996 and R(2)=0.7749) depicting lipophilicity (log P(oct)), solvent accessible molecular volume (SV), molar refractivity (MR) and system energy as the prime determinants of the brain-blood profile for these amines. The inherently high lipophilicity potential within the series is attributed to strong hydrophobic influences dominating hydrogen bonding effects. A possible conformational and energy dependent preference at the site of permeation is also suggested. The proposed estimations allow for the expedient and reliable prediction of brain partitioning behaviour for related polycyclic amines, facilitating the early rejection of unsuitable candidates and enabling research to focus on neuroprotective activity. PMID- 12901902 TI - Analogues of the mycobacterial arabinogalactan linkage disaccharide as cell wall biosynthesis inhibitors. AB - The mycobacterial arabinogalactan linkage disaccharide [alpha-L-Rha-(1-->3)-alpha D-GlcNAc] provides a basis for the design of new antitubercular drugs, since it supports a key skeletal structure in the bacterial cell wall. A series of analogues of the linker was synthesized by coupling appropriate thiorhamnosyl donors modified at their 4-positions, with an N-acetyl glucosamine acceptor. In a cell-free enzyme inhibition assay, three analogues inhibited the activity of the galactosyltransferase that adds a Galf residue to the linkage disaccharide. Although the compounds were modest inhibitors, these data confirm the viability of this approach to anti-mycobacterial agents. It is especially significant that the three effective compounds are modified at the site of the acceptor atom in the natural substrate. PMID- 12901903 TI - Caffeoyl naphthalenesulfonamide derivatives as HIV integrase inhibitors. AB - HIV-1 integrase (IN) is an essential enzyme for retroviral replication and a rational target for the design of anti-AIDS drugs. In the present study, we have designed, synthesized and tested a series of caffeoyl naphthalenesulfonamide derivatives as HIV integrase inhibitors. Among these compounds, we found that HIV integrase inhibitory activities of compounds III-3 and III-4 were more potent than L-chicoric acid (IC(50)=11.8 microg/mL) and others were comparable to L chicoric acid. Furthermore, the structure-activity relationships of these compounds were studied. The information gathered from this paper will be useful in the development and design of HIV-1 integrase inhibitors in the future. PMID- 12901904 TI - Oxidative cleavage of plasmid bluescript by water-soluble Mn-porphyrins and artificial oxidants or molecular oxygen. AB - A set of eight Mn(III)-porphyrins were used as catalysts in oxidative demolition of Plasmid Bluescript, to the nicked and linear forms, in the presence of different oxygen donors (NaOCl, H(2)O(2), AcOOH, t-BuOOH). The efficiency of the catalytic system is related to a combination of factors such as porphyrin structures, pH of the aqueous phase and nature of the primary oxidant. The highest catalytic activity was observed when ionic porphyrins were used as catalyst (the cationic being more active than the anionic) and NaOCl was used as primary oxidant at pH 9.5; in contrast, lipophilic catalysts proved to be completely unreactive towards the DNA, whichever oxidant used. The plasmid demolition was also achieved by irradiating the reaction mixture, containing Zinc porphyrins, with a white lamp; under these conditions, the highest efficiency was again observed with meso-tetra(1-methyl-4-pyridyl)porphyrin. However, preliminary experiments of photo activation applied on tumour cells (HCT 116) showed no dead cells with cationic porphyrin, while the amphiphilic Zn-tetra(4 hydroxyphenyl)porphyrin gave IC(50) values at 5 x 10(-2) microM concentration (37.1 ng/mL). PMID- 12901905 TI - 2,4-Decadienals are produced via (R)-11-HPITE from arachidonic acid in marine green alga Ulva conglobata. AB - Marine green alga Ulva conglobata was investigated for the biogeneration of oxygenated products from exogenously added arachidonic acid (ARA). A crude enzyme from the alga afforded the detectable amount of a hydroperoxyicosatetraenoic acid (HPITE), which was identified as (R)-11-HPITE by HPLC and GC-MS. Headspace-SPME method indicated that ARA was selectively used to form 2,4-decadienals. These results showed that 2,4-decadienals are produced via (R)-11-HPITE from ARA exclusively. PMID- 12901906 TI - Conformationally rigid N-acyl-5-alkyl-L-prolyl-pyrrolidines as prolyl oligopeptidase inhibitors. AB - In the N-acyl-L-prolyl-pyrrolidine type of prolyl oligopeptidase inhibitors the L prolyl group was replaced by different 5-alkyl-L-prolyl groups, resulting in a series of N-acyl-5-alkyl-L-prolyl-pyrrolidines. Since N-amides of 5-alkyl-L prolines are conformationally more rigid than those of L-proline, the main objective was to make more rigid prolyl oligopeptidase inhibitors. In the series of compounds where the N-acyl group was a Boc group, the 5(R)-tert-butyl group increased the potency strongly. A similar effect was not observed for the 5(S) tert-butyl group. In the series of compounds where the N-acyl group was a 4 phenylbutanoyl group, the 5(R)-tert-butyl, 5(R)-methyl and 5(S)-methyl groups did not have an effect on the potency [the 5(S)-tert-butyl group was not tested in this series]. As an additional effect, the 5-tert-butyl groups increased the log P of the compounds 1.5 log units, which might be beneficial when targeting the compounds to the brain. PMID- 12901907 TI - Design and synthesis of potent vitamin D receptor antagonists with A-ring modifications: remarkable effects of 2alpha-methyl introduction on antagonistic activity. AB - Novel A-ring analogues of the vitamin D receptor (VDR) antagonist (3a), ZK 159222, and its 24-epimer (3b) were convergently synthesized. Preparation of the CD-ring portions with the side chains of 3a,b, followed by palladium-catalyzed cross-coupling with the A-ring enyne precursors (15a,b), (3S,4S,5R)- and (3S,4S,5S)-bis[(tert-butyldimethylsilyl)oxy]-4-methyloct-1-en-7-yne, afforded the 2alpha-methyl-introduced analogues (4a,b) and their 3-epimers (5a,b). The biological profiles of the hybrid analogues were assessed in terms of affinity for VDR, and antagonistic activity to inhibit HL-60 cell differentiation induced by the natural hormone, 1alpha,25-dihydroxyvitamin D(3). The analogue 4a showed an approximately fivefold higher antagonistic activity compared with 3a. The 2alpha-methyl introduction into 3a increased the receptor affinity, thereby enhancing VDR antagonism. This approach to design potent antagonists based on hybridization of structural motifs in the A-ring and in the side chain may prove to be valuable. PMID- 12901908 TI - Synthesis of 1,2,3-triazolo-carbanucleoside analogues of ribavirin targeting an HCV in replicon. AB - The synthesis of carbocyclic and phosphonocarbocyclic analogues of ribavirin, an anti-HCV inhibitor, are described. Those compounds were evaluated against HCV but also against other important viruses in order to determine their spectrum of antiviral activity. Compounds 6 and 13 displayed a moderate IC(50) against HIV-1 of 43.8 and 37 microM, respectively. PMID- 12901909 TI - Synthesis and structure-activity relationships of 2-substituted-8-hydroxyadenine derivatives as orally available interferon inducers without emetic side effects. AB - Recently we reported the adenine derivatives (2-4) as new interferon (IFN) inducers. In the present study, we conducted a detailed structure and activity relationship study of 4 and its related derivatives on IFN inducing activity. From this study, we found that compound 4 exhibited the most potent IFN inducing activity in vitro with a minimum effective concentration of 0.01 microM, and 4 also showed strong IFN-inducing activity at doses of more than 0.3mg/kg by oral administration in mice. This potency was 10-fold stronger than that of Imiquimod. Moreover, 4 did not cause emesis in ferrets even at doses as high as 10mg/kg, whereas, 80% of animals were emetic when orally administered with the same dose of Imiquimod. These results indicate that compound 4 is superior to Imiquimod with respect to efficacy and safety. PMID- 12901910 TI - Synthesis and binding affinities of fluoroalkylated raloxifenes. AB - Three fluoroalkylated derivatives (1-3) of the selective estrogen receptor modulator (SERM), raloxifene, have been synthesized. The key step in the synthesis is the C-C bond formation of benzo[b]thiophene and a substituted phenyl group (ring C) using a Stille reaction. The in vitro binding affinities of the substituted raloxifenes 1-3 are 45, 60, 89%, respectively, relative to the affinity of estradiol, which is higher than the affinity of raloxifene itself (25%). When labeled with the positron-emitting radionuclide, these compounds might be useful as PET imaging agents for estrogen receptor-positive breast tumors. PMID- 12901911 TI - Cytotoxic versus anti-inflammatory effects in HeLa, Jurkat T and human peripheral blood cells caused by guaianolide-type sesquiterpene lactones. AB - Four guaianolide type sesquiterpene lactones (SL), namely the new 1,2-dihydro-3 oxo-costic acid guaianyl ester 3beta-O-(1,2-didehydro-3-oxo-costoyloxy) 4beta,10beta-dihydroxy-guaia-1(2)-en-6beta,12-olide (1) and 3beta-O-(1,2 didehydro-3-oxo-costoyloxy)-4beta,10beta-dihydroxy-guaia-1(2)-en-6alpha,12-olide (2), as well as the known moroccolide A [5alphaH-2beta,4-epoxy-3beta-hydroxy guaia-1(10),11(13)-dien-6beta,12-olide, 3] and 3beta-O-(2-methylbutyryl) moroccolide A [5alphaH-2beta,4-epoxy-3beta-(2-methylbutyryloxy)-guaia 1(10),11(13)-dien-6beta,12-olide, 4] were examined for their cytotoxic and anti inflammatory effects in HeLa, Jurkat T and human peripheral blood mononuclear cells. Compounds 1, 2 and 4 were found to exert a strong cytotoxicity similar in potency in all investigated cell types, whereas 3 was significantly less active. Along with the cytotoxic effect compounds 1 and 4 showed a potent and comparable down-regulation of the mRNAs of the house-keeping genes beta-actin and GAP-DH in PBMCs after 20 h. In contrast, the down-regulation of the PMA-induced mRNA levels of the NF-kappaB-driven pro-inflammatory genes IL-2, IL-6, GM-CSF, TNF-alpha, and IL-1beta in PBMCs is significantly stronger with compound 4. Compound 3 did not significantly modulate cytokine mRNAs levels at biochemically relevant concentrations. The electromobility shift assay (EMSA), revealed a stronger inhibition of NF-kappaB for 1 (IC(50) 2.5 microM) than for 4 (IC(50) 5 microM). Both compounds were also subjected to an IL-6 luciferase reporter gene assay and showed IC(50) values of 1.0 (1) and 1.2 microM (4). Thus, the NF-kappaB inhibition measured by EMSA, as well as the IL-6 luciferase assay did not reflect the differential modulation of pro-inflammatory genes measured with RT-rt-PCR. PMID- 12901912 TI - Allylmagnolol, a novel magnolol derivative as potent antioxidant. AB - We reported the discovery of potent antioxidants based on magnolol, a naturally occurring biphenolic obtained from the bark of Magnolia officinalis. The allylmagnolols 3a,b were synthesized via O-alkylation of the biphenols followed by Claisen rearrangement. In-vitro using enhanced chemiluminescence (CL) and flow cytometric assays in whole cells revealed that both 3a and 3b displayed promising free radical scavenging effects in PMA- and LPS-stimulated models as compared with magnolol. Further DNA labeling analysis for cytotoxicity indicated that these analogues show no cytotoxic effects for the scavenging of the oxygen derived free radicals under PMA-stimulated concentrations. The results from 3,3' bisallylmagnolol (3b) suggested that the naturally occurring constituent was suitable to be a lead compound for the development of potential antioxidants for certain diseases. PMID- 12901913 TI - Synthesis, structural characterization and in vitro antitumor activity of novel 6 chloro-1,1-dioxo-1,4,2-benzodithiazie derivatives. AB - A series of nonconventional aminium N-(6-chloro-7-R-1,1-dioxo-1,4,2 benzodithiazin-3-yl)arylsulfonamidates 7-15 have been synthesized by the reactions of 6-chloro-7-R-3-methylthio-1,4,2-benzodithiazine 1,1-dioxides with 4 dimethylaminopyridine or Et(3)N and some arylsulfonamides. The free N-(6-chloro-7 methyl-1,1-dioxo-1,4,2-benzodithiazin-3-yl)benzenesulfonamides 16-18 were obtained by treatment of their aminium salts with H(2)SO(4) in boiling acetic acid. The in vitro antitumor activity of the compounds 9, 11-14 and 16-18 has been tested in the antitumor screening of the National Cancer Institute (NCI), and relationships between structure and antitumor activity are discussed. 4 Dimethylaminopyridinium 4-chloro-N-(6-chloro-7-methyl-1,1-dioxo-1,4,2 benzodithiazin-3-yl)benzenesulfonamidate 9 is the prominent of the compounds due to its remarkable activity (log GI(50)<-8.00, log TGI=-5.50) and selectivity for the leukemia SR cell line. For that reason experimental and theoretical analysis of the geometric and electronic properties of 9 was carried out. PMID- 12901914 TI - Benzofuranyl 3,5-bis-polyamine derivatives as time-dependent inhibitors of trypanothione reductase. AB - The synthesis and evaluation of 3,5-disubstituted benzofuran derivatives as time dependent inhibitors of the protozoan oxidoreductase trypanothione reductase are reported. These molecules were designed as simplified mimetics of the naturally occurring spermidine-bridged macrocyclic alkaloid lunarine 1, a known time dependent inhibitor of trypanothione reductase. In this series of compounds the bis-polyaminoacrylamide derivatives 2-4 were all shown to be competitive inhibitors, but only the bis-4-methyl-piperazin-1-yl-propylacrylamide derivative 4 displayed time-dependent activity. The kinetics of time dependent inactivation of trypanothione reductase by 1 and 4 have been determined and are compared and discussed herein. PMID- 12901915 TI - Design and synthesis of poly(ADP-ribose) polymerase-1 (PARP-1) inhibitors. Part 4: biological evaluation of imidazobenzodiazepines as potent PARP-1 inhibitors for treatment of ischemic injuries. AB - A class of poly(ADP-ribose) polymerase (PARP-1) inhibitors, the imidazobenzodiazepines, are presented in this text. Several derivatives were designed and synthesized with ionizable groups (i.e., tertiary amines) in order to promote the desired pharmaceutical characteristics for administration in ischemic injury. Within this series, several compounds have excellent in vitro potency and our computational models accurately justify the structure-activity relationships (SARs) and highlight essential hydrogen bonding residues and hydrophobic pockets within the catalytic domain of PARP-1. Administration of these compounds (5q, 17a and 17e) in the mouse model of streptozotocin-induced diabetes results in maintainance of glucose levels. Furthermore, one such inhibitor (5g, IC(50)=26 nM) demonstrated significant reduction of infarct volume in the rat model of permanent focal cerebral ischemia. PMID- 12901916 TI - Rebeccamycin analogues bearing amine substituents or other groups on the sugar moiety. AB - In the course of structure-activity relationship studies on rebeccamycin analogues, a series of compounds bearing an amino function on the sugar moiety were synthesized with the aim of improving the solubility and interaction with the macromolecular target(s). The syntheses of amino derivatives and the corresponding chloro, iodo and azido intermediates are described. Their interaction with DNA and effects on human DNA topoisomerases I and II were investigated. Their antimicrobial activities against two Gram-positive bacteria, Bacillus cereus and Streptomyces chartreusis, a Gram-negative bacterium Escherichia coli and a yeast Candida albicans were also determined. 6'-Amino compound 7 and 6'-N-methylamino 14 very efficiently inhibit the growth of E. coli. The introduction of an amino group at the 6'-position strongly enhances the capacity of the drugs to interact with DNA but almost abolishes their poisoning effect on topoisomerase I. Unlike the vast majority of rebeccamycin analogues previously studied, the newly designed compounds do not stimulate DNA cleavage by topoisomerase I. The enhanced capacity of the 6'-amino glycosyl rebeccamycin derivatives to bind to DNA likely account for the improved biological profiles. DNA and topoisomerase I represent two independent targets which can both be used for the development of antitumor rebeccamycin derivatives. PMID- 12901917 TI - C2-symmetric inhibitors of Plasmodium falciparum plasmepsin II: synthesis and theoretical predictions. AB - A series of C(2)-symmetric compounds with a mannitol-based scaffold has been investigated, both theoretically and experimentally, as Plm II inhibitors. Four different stereoisomers with either benzyloxy or allyloxy P1/P1' side chains were studied. Computational ranking of the binding affinities of the eight compounds was carried out using the linear interaction energy (LIE) method relying on a complex previously determined by crystallography. Within both series of isomers the theoretical binding energies were in agreement with the enzymatic measurements, illustrating the power of the LIE method for the prediction of ligand affinities prior to synthesis. The structural models of the enzyme inhibitor complexes obtained from the MD simulations provided a basis for interpretation of further structure-activity relationships. Hence, the affinity of a structurally similar ligand, but with a different P2/P2' substituent was examined using the same procedure. The predicted improvement in binding constant agreed well with experimental results. PMID- 12901919 TI - P-loop catalytically assisting the enzymatic cleavage of single-stranded DNA. AB - We demonstrated that a P-loop, a looped complex formed inside duplex DNA by adding peptide nucleic acids (PNA), acts catalytically as a template for enzymatic cleavage of single-stranded probe oligodeoxynucleotides (ODN). A PD loop complex formed from P-loop and probe ODN was digested efficiently by a restriction enzyme, and the truncated probe ODN was released. The P-loop nicked by the enzyme can form PD-loop again with another probe ODN, and then assisted the enzymatic cleavage of an excess of probe ODN. In addition, by using dumbbell formed ODN as a probe ODN, the efficiency of the P-loop-assisted ODN cleavage was enhanced considerably as compared with that of linear ODN. Thus, the method utilizing P-loop will make it possible to amplify the sequence information of duplex DNA via a catalytic cleavage of probe ODNs. PMID- 12901918 TI - Syntheses of 3-ethylidenequinuclidine derivatives as squalene synthase inhibitors. Part 2: enzyme inhibition and effects on plasma lipid levels. AB - Squalene synthase (E.C. 2.5.1.21) is a microsomal enzyme which catalyzes the reductive dimerization of two molecules of farnesyl diphosphate to form squalene, and is involved in the first committed step in cholesterol biosynthesis. It is an attractive target for hypocholesterolemic and hypotriglyceridemic strategies. We synthesized a series of 3-ethylidenequinuclidine derivatives, and evaluated their ability to inhibit squalene synthase in vitro and to lower non-HDL cholesterol levels in hamsters. 3-Ethylidenequinuclidine derivatives incorporating an unsubstituted 9H-carbazole moiety reduced plasma non-HDL cholesterol levels and did not affect plasma transaminase levels, indicating a lack of hepatotoxicity. Among the novel compounds, (Z)-2-[2-(quinuclidin-3-ylidene)ethoxy]-9H-carbazole hydrochloride 8 (YM-53579) and (E)-2-[2-fluoro-2-(quinuclidin-3-ylidene)ethoxy] 9H-carbazole hydrochloride 28 (YM-53601) were potent inhibitors of squalene synthase derived from human hepatoma cells, with IC(50) values of 160 and 79 nM, respectively. They also reduced plasma non-HDL cholesterol levels in hamsters by approximately 50 and 70%, respectively, at an oral dose of 50 mg/kg/day for 5 days. PMID- 12901920 TI - Three-dimensional molecular-field analyses of octopaminergic agonists for the cockroach neuronal octopamine receptor. AB - The quantitative structure-activity relationship of a set of 40 octopaminergic agonists against receptor 2 in cockroach nervous tissue, was analyzed using molecular-field analysis (MFA). MFA on the study set of those compounds evaluated effectively the energy between a probe and a molecular model at a series of points defined by a rectangular grid. Contour surfaces for the molecular fields were presented and the results provided useful information in the characterization and differentiation of octopaminergic receptor. PMID- 12901921 TI - pKa and volume of residue one influence delta/mu opioid binding: QSAR analysis of tyrosine replacement in a nonselective deltorphin analogue. AB - [Gly(4)]deltorphin (Tyr-D-Ala-Phe-Gly-Val-Val-Gly-NH(2)) is a nonselective analogue of the opioid heptapeptides isolated from Phyllomedusa amphibian skin. Its nonselective nature allows for simultaneous characterization of the effects of sequence modification on both delta (delta) and mu (mu) receptor binding. The N-terminal regions of opioid peptides are considered to be responsible for receptor recognition, and the tyrosine at position one is relatively intolerant to alteration. In order to further investigate the role of the phenolic hydroxyl group in receptor interaction, a series of peptides was synthesized in which the position-one tyrosine residue was replaced with analogues of varying electronic, steric, and acid/base character, including ring-substituted tyrosines, para substituted phenylalanines, and other nonaromatic and heterocyclic amino acids. The effects of these replacements on delta and mu receptor affinities were measured and then analyzed through quantitative structure-activity relationship (QSAR) calculations. Results support a dual hydrogen bond donor/acceptor role for the Tyr(1) hydroxyl moiety, with less acidic hydroxyl groups exhibiting stronger binding to opioid receptors. In addition, steric bulk in the Tyr(1) position independently strengthens mu and possibly delta binding, presumably by either a ligand conformational effect or enhanced van der Waals interactions with a 'loose' receptor site. The pK(a) effect is stronger on delta than on mu binding, generating an increase in delta selectivity with increasing residue-one pK(a). PMID- 12901922 TI - New benzo[g]isoquinoline-5,10-diones and dihydrothieno [2,3-b]naphtho-4,9-dione derivatives: synthesis and biological evaluation as potential antitumoral agents. AB - Novel antitumoral agents with quinonic structure were synthesized and evaluated for their in vitro cytotoxic activities. This study examines the cytotoxic activities of several aryl benzo[g]isoquinoline-5,10-dione derivatives and a number of aminoacyl dihydrothieno[2,3-b]naphtho-4,9-dione (DTNQ) derivatives containing amino acids in position 3 of the ring system. Compound 6 showed remarkable cytotoxic activity at submicromolar concentration not only against several human leukaemia and solid tumour cell lines, but also toward sensitive and resistant human cell lines. PMID- 12901923 TI - Synthesis and structure-activity relationship of 2-amino-3-heteroaryl quinoxalines as non-peptide, small-molecule antagonists for interleukin-8 receptor. AB - Interleukin-8 modulation is implicated in many inflammatory and cancer diseases. Starting from a mass-screening hit, the synthesis and structure-activity relationship of 2-amino-3-heteroarylquinoxalines as non-peptide, small molecule interleukine-8 receptor antagonists have been developed. The optimized derivatives, PD 0210293 (13y) and PD 0220245 (13r), show inhibition of both IL-8 receptor binding and IL-8-mediated neutrophil chemotaxis. PMID- 12901924 TI - A short synthesis of antimalarial peroxides. AB - A concise and efficient synthesis of two simplified diastereomeric analogues of a natural peroxide is presented. Both compounds could be isolated in high purity and fully identified. They exhibited moderate antimalarial activity. PMID- 12901925 TI - 8,9-methylenedioxybenzo[i]phenanthridines: topoisomerase I-targeting activity and cytotoxicity. AB - Substituted benzo[i]phenanthridines that have incorporated within their structure an 8,9-methylenedioxy group can exhibit topoisomerase I-targeting activity. Structure-activity studies were performed to examine the influence of saturation at the 11,12-positions of several substituted 8,9 methylenedioxybenzo[i]phenanthridines. The activities of these dihydro analogues were compared to those of their unsaturated analogues. In addition, the influence of varying substituents at the 2- and 3-positions within the A-ring of these 8,9 methylenedioxybenzo[i]phenanthridines on their relative potency as topoisomerase I-targeting agents and cell proliferation as determined using the MTT assay was investigated. 2,3-Dimethoxy-8,9-methylenedioxybenzo[i]phenanthridine and its 11,12-dihydro derivative were among the more potent analogues evaluated with regard to topoisomerase I-targeting activity and cytotoxicity. PMID- 12901926 TI - Synthesis of amide compounds of ferulic acid, and their stimulatory effects on insulin secretion in vitro. AB - We prepared amide compounds which were derived from ferulic acid using various amines, and investigated their stimulatory effects on insulin secretion using rat pancreatic RIN-5F cells. Most of these compounds exhibited significant promotion of the insulin-release at a concentration of 10 microM and in particular, the amides having n-butyl, n-pentyl, pyrrolidine, and piperidine groups showed high activity. PMID- 12901927 TI - Establishment of substituent effects in the DNA binding subunit of CBI analogues of the duocarmycins and CC-1065. AB - An extensive series of CBI analogues of the duocarmycins and CC-1065 exploring substituent effects within the first indole DNA binding subunit is detailed. In general, substitution at the indole C5 position led to cytotoxic potency enhancements that can be >/=1000-fold providing simplified analogues containing a single DNA binding subunit that are more potent (IC(50)=2-3 pM) than CBI-TMI, duocarmycin SA, or CC-1065. PMID- 12901928 TI - Efficacy validation of synthesized retinol derivatives In vitro: stability, toxicity, and activity. AB - Retinol (vitamin A) is used as an antiwrinkle agent in the cosmetics industry. However, its photo-instability makes it unsuitable for use in general cosmetic formulations. To improve the photo-stability of retinol, three derivatives (3, 4, and 5) were synthesized and their biological activities were analyzed. 1H NMR and HPLC analysis indicated that derivatives 3 and 5 were much more stable than retinol under our sunlight exposure conditions. When human adult fibroblasts were treated, the IC(50) of derivative 3 was 96 microM, which is similar to that of retinol, as determined by the MTT assay. Derivatives 4 and 5 were 2.5 and 8 times more toxic than retinol, respectively. At 1 microM treatment, like retinol, derivatives 3 and 4 were specifically active for RARalpha out of six retinoid receptors (RAR/RXRalpha, beta, gamma). Dose-dependent analysis confirmed that derivative 4 was as active as retinol and the other two derivatives were less active for RARalpha. The effect of our derivatives on the expression of collagenase, an indicator of wrinkle formation, was measured using the transient co-expression of c-Jun and RT-PCR in HaCaT cells. Collagenase promoter activity, which is increased by c-Jun expression, was reduced 42% by retinol treatment. The other derivatives inhibited collagenase promoter activity similarly. These results were further confirmed by RT-PCR analysis of the collagenase gene. Taken together, our results suggest that retinol derivative 3 is a promising antiwrinkle agent based on its higher photo-stability, lower RARalpha activity (possibly indicating reduced side effects), and similar effect on collagenase expression. PMID- 12901929 TI - The mosquito genome--a turning point? AB - Of the insects that serve as vectors for parasitic diseases, the genus Anopheles is the most important. Of the approximately 400 species, about twelve serve as vectors for human pathogens. Months have passed since the sequenced genomes of the malaria parasite, Plasmodium falciparum, and its vector, Anopheles gambiae, were published. Sequences were compared, gene and protein predictions were made, new research areas evolved and many ongoing projects gained new momentum. A general belief is that we are at a turning point: we are now in a position to tackle both the parasite and the vector from new angles and with new force, for example, by identifying new drug targets and obtaining a deeper insight into molecular mechanisms of the insect, the parasite and the interactions between them. PMID- 12901930 TI - Amphibian deformities and Ribeiroia infection: an emerging helminthiasis. AB - Since their widespread appearance in the mid-1990s, malformed amphibians have evoked fear, as well as fascination within the scientific and public communities. Recent evidence from field and laboratory studies has implicated infection by a digenetic trematode--Ribeiroia ondatrae--as an important cause of such deformities. Ribeiroia spp. have a complex life cycle involving planorbid snails, amphibians and water birds. Under natural conditions, malformations might promote parasite transmission by increasing the susceptibility of infected amphibians to predation by definitive hosts. However, with respect to the recent outbreak of deformities, we suggest that exogenous agents (e.g. pesticides, nutrient run-off, introduced fishes) might be interacting with Ribeiroia, resulting in elevated infection levels, and we highlight the need for studies incorporating multiple stressor dynamics to further explore this problem. PMID- 12901931 TI - Genes, nitric oxide and malaria in African children. AB - The unresolved and complex relationship between nitric oxide and falciparum malaria is reflected in recent genetic and immunohistochemical studies in African children. Different genetic associations, perhaps geographically distinctive, are seen between genetic variants of the inducible nitric oxide gene and various disease manifestations in African populations. The picture might not be complete without considering the emerging roles of carbon monoxide, another endogenous gaseous mediator with similar effects to those of nitric oxide. Only when genetic comparisons from across tropical Africa are examined, in conjunction with the newly recognized complexities in the events of systemic inflammation, will this relationship be understood. PMID- 12901932 TI - Trichinella-infected pork products: a dangerous gift. PMID- 12901933 TI - Invasion by schistosome cercariae: studies with human skin explants. PMID- 12901934 TI - Integrated control of vector-borne diseases of livestock--pyrethroids: panacea or poison? AB - Tick- and tsetse-borne diseases cost Africa approximately US$4-5 billion per year in livestock production-associated losses. The use of pyrethroid-treated cattle to control ticks and tsetse promises to be an increasingly important tool to counter this loss. However, uncontrolled use of this technology might lead to environmental damage, acaricide resistance in tick populations and a possible exacerbation of tick-borne diseases. Recent research to identify, quantify and to develop strategies to avoid these effects are highlighted. PMID- 12901935 TI - Importance and pitfalls of molecular analysis to parasite epidemiology. AB - Molecular tools are increasingly being used to address questions about parasite epidemiology. Parasites represent a diverse group and they might not fit traditional population genetic models. Testing hypotheses depends equally on correct sampling, appropriate tool and/or marker choice, appropriate analysis and careful interpretation. All methods of analysis make assumptions which, if violated, make the results invalid. Some guidelines to avoid common pitfalls are offered here. PMID- 12901936 TI - The first releases of transgenic mosquitoes: an argument for the sterile insect technique. AB - Potential applications for reducing transmission of mosquito-borne diseases by releasing genetically modified mosquitoes have been proposed, and mosquitoes are being created with such an application in mind in several laboratories. The use of the sterile insect technique (SIT) provides a safe programme in which production, release and mating competitiveness questions related to mass-reared genetically modified mosquitoes could be answered. It also provides a reversible effect that would be difficult to accomplish with gene introgression approaches. Could new technologies, including recombinant DNA techniques, have improved the success of previous mosquito releases? Criteria for an acceptable transgenic sterile mosquito are described, and the characteristics of radiation-induced sterility are compared with that of current transgenic approaches. We argue that SIT using transgenic material would provide an essentially safe and efficacious foundation for other possible approaches that are more ambitious. PMID- 12901937 TI - Malaria parasite and vector genomes: partners in crime. AB - The publication of the genome sequences of the malaria parasite Plasmodium falciparum and the insect vector Anopheles gambiae paves the way for scientists to study these organisms by using technologies developed to observe global changes in transcription and translation, as well as computational tools. Researchers are now able to investigate complex changes involved in development, growth and reaction to external factors. Given the medical importance of these organisms, much of this work is targeted on drug or insecticide discovery (including mechanisms of resistance to existing treatments), but the genome information also provides the opportunity to develop novel therapies. PMID- 12901938 TI - Estimating the needs for artesunate-based combination therapy for malaria case management in Africa. AB - Because of inadequacies in national health information systems, the volumes of drugs required to support an effective policy transition toward artesunate-based combination therapy (ACT) are unknown for most African countries. A series of national surveys and population projections have been used to estimate the age structured fever burden among 41 malaria endemic countries in Africa. Under present fever-management guidelines, commodity costs and internationally agreed coverage targets, the financial resources to meet the needs of ACT in most African countries are huge. Between US$1.6 billion and US$3.4 billion per annum must be found to give Africa the chance to consider a drug policy based on ACT. Substantial reductions in these costs would be achieved through more effective targeting of resources--only 20% of drugs would be required to manage fevers among the most at-risk pediatric patient populations. Better diagnostics would also be an important consideration for a new ACT policy in Africa. PMID- 12901939 TI - Mixed models: getting the best use of parasitological data. AB - Statistical analysis of parasitological data provides a powerful method for understanding the biological processes underlying parasite infection. However, robust and reliable analysis of parasitological data from natural and experimental infections is often difficult where: (1) the distribution of parasites between hosts is aggregated; (2) multiple measurements are made on the same individual host in longitudinal studies; or (3) data are from 'noisy' natural systems. Mixed models, which allow multiple error terms, provide an excellent opportunity to overcome these problems, and their application to the analysis of various types of parasitological data are reviewed here. PMID- 12901940 TI - The bullseye of cancer therapy: a moving target. PMID- 12901941 TI - Differentiation therapy. AB - Differentiation therapy is an area of oncology that is in its infancy. Theoretically, the concept of differentiation therapy involves turning a cancer cell "off" biologically and reverting to a more "benign" phenotype. Many agents have been studied over the past few years, with many already either in use clinically or showing future promise. PMID- 12901942 TI - Histone deacetylases. AB - Post-translational modification of the histones of chromatin has a fundamental role in regulating gene expression. Enzymes involved in these epigenetic events include histone deacetylases (class I and class II), which can be inhibited by a structurally diverse group of small molecules. These histone deacetylase inhibitors induce growth arrest, differentiation and/or apoptosis of cancer cells in vitro and in vivo. Results of clinical trials with several of these agents have indicated that they are well tolerated at doses that have anti-tumour activity. PMID- 12901943 TI - Cyclooxygenase inhibitors: drugs for cancer prevention. AB - Evidence that chronic intake of non-steroidal anti-inflammatory drugs, especially aspirin, prevents cancer development continues to accumulate. The data are particularly convincing for colorectal cancer; however, because of well-known side effects, they cannot routinely be recommended for this purpose. An appreciation of the mechanisms that underlie their anti-cancer effects might permit the development of safer agents. Intensive investigation has led to the characterization of several potential chemopreventive mechanisms of action of these drugs. Antineoplastic actions could result from effects on overlapping processes in the different cell-types that comprise tumors, such as epithelial and stromal cells. PMID- 12901944 TI - Cyclin-dependent kinase inhibitors. AB - Cell-cycle dysregulation is one of the cardinal characteristics of neoplastic cells. For this reason, small molecule inhibitors targeting cyclin-dependent kinases (CDKs), of which flavopiridol is a prototype, have been the focus of extensive interest in cancer therapy. In addition to inhibiting cell-cycle progression, these agents exhibit a variety of other activities, including the induction of cell death. Recently, several novel mechanisms of action have been ascribed to the CDK inhibitor flavopiridol, including interference with transcription, most likely through disruption of P-TEFb (i.e. the CDK9/cyclin T complex), and induction of apoptosis, possibly a consequence of downregulation of various anti-apoptotic proteins. It has also been observed that combining CDK inhibitors with either conventional cytotoxic drugs or novel signal transduction modulators dramatically promotes neoplastic cell death in a variety of preclinical models. Efforts are underway to uncover inhibitors that selectively target specific CDKs and to develop these as a new generation of antitumour drugs. For all of these reasons, it is likely that interest in CDK inhibitors as antineoplastic agents will continue for the foreseeable future. PMID- 12901945 TI - Targeting mTOR signaling for cancer therapy. AB - The mammalian target of rapamycin (mTOR), an atypical serine/threonine kinase, plays a central role in the regulation of cell proliferation, growth, differentiation, migration and survival. Dysregulation of mTOR signaling occurs in diverse human tumours, and can confer higher susceptibility to inhibitors of mTOR. Rapamycin and its derivatives, CCI-779 and RAD001 (designated rapamycins), specifically inhibit the function of mTOR, leading to inactivation of ribosomal S6K1 and inhibition of cap-dependent translation initiation through the 4E BP1/eIF4E pathway. The overall effect is an accumulation of cells in the G1 phase of the cell-cycle, and potential apoptosis. Preclinical studies indicate that rapamycins are potent inhibitors of the proliferation of numerous tumour cell lines in culture and of murine syngeneic tumour models or human xenografts. RAD001 and CCI-779 are in phase I and II trials, respectively, as anti-cancer agents. These trials have demonstrated promising anti-cancer activity and relatively mild side effects of CCI-779. Emerging results suggest that inhibition of mTOR signaling can be exploited as a potential tumour-selective therapeutic strategy. PMID- 12901946 TI - Tailoring chemotherapy in advanced colorectal cancer. AB - In this post-genomic era, the individualization of chemotherapy through the study of pharmacogenetics is becoming an ever attainable reality. Keys to individual variations in drug response and toxicity are being identified through the study of drug targets, metabolizing enzymes, efflux and DNA repair systems at the genomic, mRNA and protein levels. Several promising pharmacogenetic candidates with predictive and/or prognostic value have been identified. These candidates, along with others yet to be identified, could in the near future allow for the tailoring of therapy with an ever increasing chemotherapeutic armamentarium. Well designed and large prospective analyses, which include relevant pharmacogenetic parameters, are needed to confirm the initial clinical associations reported thus far. PMID- 12901947 TI - Antibody-targeted chemotherapy with immunoconjugates of calicheamicin. AB - Targeted delivery of cytotoxic agents to tumours is believed to improve both their anti-tumour efficacy and their safety. Antibodies specific for tumour associated antigens have been used to deliver cytotoxic agents to tumour cells. Calicheamicin is a potent cytotoxic agent that causes double-strand DNA breaks, resulting in cell death. When conjugated to monoclonal antibodies specific for tumour-associated antigens, calicheamicin exerts strong antigen-specific anti tumour effects against human tumour xenografts in preclinical models. Antibody targeted chemotherapy with immunoconjugates of calicheamicin, exemplified by gemtuzumab ozogamicin (Mylotarg), is a clinically validated therapeutic strategy for the treatment of human cancer. PMID- 12901948 TI - Spotlight on new anti-inflammatory drug targets in the immune system. PMID- 12901949 TI - Therapeutic targeting of Toll-like receptors for inflammatory and infectious diseases. AB - Roles for Toll-like receptors (TLRs) are emerging in conditions such as sepsis syndrome, systemic lupus erythromatosis, rheumatoid arthritis and asthma, suggesting that the selective targeting of TLRs might be useful therapeutically. TLRs are defined by the presence of extracellular leucine-rich repeats and an intracellular Toll/interleukin-1 receptor domain, and play a role in host defence and inflammation. Signalling pathways activated by TLRs show remarkable similarity to those activated by the pro-inflammatory cytokine interleukin-1 (the receptor for which also has a Toll/interleukin-1 receptor domain), although adaptor proteins specific for certain TLRs are starting to emerge (e.g. Mal and Trif). The common signalling pathways used by all members of the TLR superfamily are being targeted, with drugs that block nuclear factor-kappaB and p38 mitogen activated protein kinase in clinical development for diseases such as rheumatoid arthritis and psoriasis. As we learn more about TLR signal transduction, more options are presenting themselves for pharmacological targeting. PMID- 12901950 TI - Crosstalk between glucocorticoids and mitogen-activated protein kinase signalling pathways. AB - Synthetic glucocorticoids are potent modulators of the immune system, and are clinically invaluable in the treatment of a wide variety of conditions. However, their use can be limited by harmful side effects, and their mechanisms of action remain poorly understood and controversial. It has recently been reported that glucocorticoids induce the expression of two genes that participate in cross-talk with the mitogen-activated protein kinase signalling pathways. It is possible that both therapeutic and harmful effects of glucocorticoids may be mediated by these genes. PMID- 12901951 TI - Targeting inflammatory diseases via apoptotic mechanisms. AB - Induction of apoptosis in immune cells is a crucial mechanism used by the body to produce immune resolution. The homeostatic mechanisms employed are currently being identified and, to date, studies have highlighted some of the signals that regulate the immune response. The exposure of phosphatidylserine on the surface of an apoptotic neutrophil is sufficient to limit the immune response in acute inflammation, whereas apoptosis of key effector cells can limit the response in chronic inflammation. Other therapeutic approaches that are being investigated include the inhibition of apoptosis by blocking the caspase cascade. This approach will be of particular relevance for the treatment of inflammatory central nervous system diseases and sepsis. An alternative approach being examined is forced resolution, whereby apoptosis is induced in effector cells, principally T cells, through activation-induced cell death mediated by Fas receptors. Inhibitors of this mechanism have been identified and targeted in several studies. PMID- 12901952 TI - JNK: a new therapeutic target for diabetes. AB - Jun N-terminal kinase (JNK) regulates the transcription factor AP-1, which is implicated in the controlled expression of many genes involved in the immune response. For this reason, drug discovery efforts have focused on the development of JNK inhibitors for chronic inflammatory diseases. However, recent genetic evidence and emerging pharmacological data indicate that activated JNK could be critical in causing diabetes, insulin resistance and obesity. Indeed, if JNK is considered as a stress-activated protein kinase, there appear to be multiple mechanisms through which it might promote diabetes. PMID- 12901953 TI - Isoform-specific phosphoinositide 3-kinase inhibitors as therapeutic agents. AB - The phosphoinositide 3-kinase (PI3K) family of enzymes consists of several closely related isoforms that are thought to have distinct biological roles. Until now, researchers have been frustrated by poor selectivity of the available pharmacological inhibitors, which are unable to distinguish adequately the activities of different PI3K isoforms. Recently published patent specifications describe new PI3K inhibitors, including several that are selective for the PI3Kdelta isoform. There is now cautious optimism that isoform-selective PI3K inhibitors will provide new avenues for therapeutic applications in a range of diseases. PMID- 12901954 TI - Negative regulation of cytokine signaling and inflammatory diseases. AB - Immune and inflammatory systems are controlled by multiple cytokines, including interleukins and interferons. These cytokines exert their biological functions through Janus tyrosine kinases and signal transducer and activators of transcription (STATs). The cytokine-inducible SH2 proteins (CIS) and suppressors of cytokine signaling (SOCS) are a family of intracellular proteins, several of which have emerged as key physiological regulators of cytokine responses, including those that regulate the inflammatory system. Several recent advances have been made in this field, including treatment of inflammatory diseases by modulating extracellular and intracellular signaling pathways. PMID- 12901955 TI - Chemokines in allergic airway disease. AB - Expression of chemokine receptors on T helper 2 cells and eosinophils has been postulated to be the mechanism by which these cells are selectively recruited to the lung during allergic inflammatory reactions. Mouse models have provided evidence to show that blocking the ligands for these receptors is successful in abrogating the pathophysiological effects of allergen challenge. However, recent studies describing the effect of genetic deletions of these chemokine receptors have not confirmed the results obtained with ligand knockouts or neutralising antibodies. Coupled with the realisation that, because of a lack of species cross reactivity, it is not possible to test small molecule antagonists against human receptors in the original in vivo animal models, the future of chemokine receptor therapeutics is in question. However, recent advances have been made regarding the therapeutic potential of blocking the chemokine receptors CCR3, CCR4 and CCR8 in allergic airway disease. PMID- 12901956 TI - T helper type-2 cytokine responses: potential therapeutic targets. AB - T helper (Th)2 cell-mediated immune responses are associated with parasitic helminth infections and atopic disorders. The production of interleukin (IL)-4, IL-5, IL-9 and/or IL-13 by Th2 cells mediates a range of responses that can be protective or pathogenic. Progress has recently been made in elucidating the mechanism of Th2 immunity, which has therapeutic potential for the treatment of allergic diseases. PMID- 12901957 TI - Showdown for Medicare reforms. PMID- 12901958 TI - Breast cancer trial with erythropoietin terminated unexpectedly. PMID- 12901959 TI - Pharmaceutical companies often provide insufficient information about genotoxicity and carcinogenicity assays. PMID- 12901960 TI - The use of surgery to treat lung cancer in elderly patients. AB - Lung cancer is a leading cause of cancer death and its cure depends on an adequate surgical approach. More than half of all lung cancers are diagnosed in patients aged 65 years or over. However, surgical risk increases in patients over 65 years old. Therefore, surgical procedures for lung cancer are far less frequent in elderly patients. Many clinicians avoid surgery, or minimise surgical procedures on the basis of age but recent advances in preoperative risk assessment and surgical and anaesthetic techniques have resulted in a significant decrease in operative mortality and morbidity for older patients. The treatment of lung cancer in elderly patients should no longer be based on the premise that surgery is too risky for elderly patients. Every effort should be made to assess risk and optimise treatment for this large and expanding proportion of the population. PMID- 12901961 TI - Neuroblastoma: biology and molecular and chromosomal pathology. AB - Neuroblastoma is the most frequently occurring solid tumour in children, with an incidence of 1.3 cases per 100000 children aged 0-14 years. Despite many advances during the past three decades, neuroblastoma has remained an enigmatic challenge to clinical and basic scientists. 20 years ago, the MYCN gene was found to be amplified in neuroblastomas, and research since then has focused on the search for other genetic markers. It has emerged that neuroblastoma cells, like cells of many other tumour types, often suffer from extensive, non-random genetic damage at multiple genetic loci. Elucidation of the exact molecular make-up of neuroblastomas will enable researchers to analyse how much specific markers, alone or in combination, can help to stratify disease in prospective studies; at present, stratification is based on age, stage, MYCN, and Shimada pathology. Neuroblastoma may be one of the first examples of the use of genetic tumour markers as a tool for defining tumour behaviour and to aid clinical staging. PMID- 12901962 TI - The role of surgery in the management of oesophageal cancer. AB - In the absence of medical contraindications to surgery, resection is the mainstay of treatment for localised oesophageal cancer. Advancements in preoperative staging and imaging, anaesthesia delivery, surgical technique, and postoperative care, now enable the surgeon to safely operate on patients with oesophageal tumours and to tailor the procedure on the basis of performance status, tumour location, and extent of disease. During the past 10 years, several "minimally invasive" techniques, which aim to limit the extent of resection, have been introduced; these procedures are currently being investigated for use in both staging and treatment of oesophageal malignant diseases. Despite these accomplishments however, overall 5-year survival remains disappointing: less than 25% of patients live for 5 years after oesophagectomy. For patients with locally or regionally advanced disease (stage IIa, IIb, III, and IVa), combining several treatment approaches, either with or without surgery, can result in good objective responses and, in some patients, durable survival. The role of surgery in such combined modality approaches is still evolving and some investigators have challenged its worth. To provide a definitive review of the issues involved, we outline the types of surgery used to treat cancer of the oesophagus and summarise the available data about their effectiveness. Clinical outcomes, the value of preoperative chemoradiotherapy, and the use of surgery are all considered. PMID- 12901963 TI - The systemic treatment of advanced and metastatic bladder cancer. AB - Bladder cancer is the second most common genitourinary tumour and is a significant cause of morbidity and mortality. Trials of neoadjuvant and adjuvant chemotherapy have failed to show a survival advantage, although these studies generally had suboptimum design and an insufficient number of patients. Despite the introduction of newer agents, the median survival for metastatic disease is about 1 year; however, improvements in quality of life have been achieved. Platinum drugs should be included in studies of combination chemotherapy regimens wherever possible. There have been various studies exploring the role of taxanes, gemcitabine, ifosfamide, and platinum in double and triple combinations in different schedules to maximise dose intensity and improve effectiveness but large phase III trials are needed. The current tumour, node, and metastasis staging system is insufficient to predict outcome in patients with bladder cancer irrespective of the treatment they received. Evaluation of molecular prognostic markers should be incorporated into phase II and III trials to define their roles in clinical outcome. Future studies should stratify patients according to the number of risk factors they have to avoid imbalance in treatment groups and patients should be carefully selected. PMID- 12901964 TI - CD1a: a novel biomarker for Barrett's metaplasia? PMID- 12901965 TI - Chemoradiation for resectable gastric cancer. AB - The incidence of gastric cancer has been declining in recent years, however, the disease continues to be a worldwide public health problem. About two thirds of patients with gastric cancer undergo surgical resection with curative intent. R0 resection--complete local-regional tumour removal with negative resection margins -is the only curative modality. The optimum extent of lymph-node dissection (D1 vs D2) is controversial. Disease relapse, both local and distant, is common and the 5-year survival rate is disappointing. Adjuvant chemotherapy has been studied extensively in this setting but an effective regimen has not yet been identified. A recent intergroup study has shown that postoperative chemoradiation is effective in improving both disease-free survival (3-year, 48% vs 31%, p<0.001) and overall survival (3-year, 50% vs 41%, p=0.005) compared with surgery alone. Preoperative radiation as a single adjuvant therapy has also yielded improvements in local-regional control, disease-free survival, and overall survival compared with surgery alone. Preoperative chemotherapy or chemoradiation has been accepted to have a theoretical advantage over postoperative therapy and has now been shown to be a feasible option. Its efficacy, however, remains to be tested. PMID- 12901967 TI - Intracranial metastasis in carcinoma of the glottis. PMID- 12901966 TI - B-cell chronic lymphocytic leukaemia: a polymorphic family unified by genomic features. AB - Human cancer is characterised by complex molecular aberrations which result in a wide variety of clinical manifestations. B-cell chronic lymphocytic leukaemia (B CLL) is particularly diverse, both in terms of molecular changes and clinical course, and consequently our understanding of the pathology of this disease is generally poor. Furthermore, the heterogeneity of this tumour type coupled with the absence of an obvious genetic "hallmark", such as gain of oncogene function or loss of suppressor-gene function, has led many investigators to question whether B-CLL is a single disease entity. In most cases, B-CLL does not show specific reciprocal chromosomal translocations as found in other haemopoietic malignant diseases. The genomic instability of B-CLL results in numerous different types of chromosomal losses and gains, giving rise to unsettled karyotypes among individuals with this disease. Nevertheless, genetic data imply that B-CLL is a single disease characterised by a common gene-expression profile and by the existence of specific subtypes that may have clinical correlates in patients. PMID- 12901969 TI - A toast to chance. PMID- 12901970 TI - The role of stem cells in aging. AB - The objectives of this review were first to critically review what is known about the effects of aging on stem cells in general, and hematopoietic stem cells in particular. Secondly, evidence is marshalled in support of the hypothesis that aging stem cells play a critical role in determining the effects of aging on organ function, and ultimately on the lifespan of a mammal. Aging has both quantitative and qualitative effects on stem cells. On balance, the qualitative changes are the more important since they affect the self-renewal potential, developmental potential, and interactions with extrinsic signals, including those from stroma. Although hematopoiesis is generally maintained at normal and life supporting levels during normal aging, diminished function is acutely apparent when old stem cells are subjected to stress. There is ample evidence of diminished self-renewal capacity, restriction of the breadth of developmental potency, and decreased numbers of progeny of old stem cells subjected to hematopoietic demands. The prediction is made that whatever plasticity in developmental potential possessed by a young stem cell is lost during aging. Those parts of the world enjoying an ever-increasing standard of living are also inhabited by an increasingly elderly population. The effects of age on many physiological functions are not well studied or appreciated. A public health challenge to provide increased quality of life for this growing segment of the population requires more attention to the variable of age in experimental studies. Stem cell populations are likely to be a fruitful subject for studies of this type. PMID- 12901971 TI - Fps/Fes and Fer protein-tyrosinekinases play redundant roles in regulating hematopoiesis. AB - OBJECTIVE: The highly related protein-tyrosine kinases Fps (also called Fes) and Fer are sole members of a subfamily of kinases. In this study, knock-in mice harboring kinase-inactivating mutations in both fps and fer alleles were used to assess functional redundancy between Fps and Fer kinases in regulating hematopoiesis. METHODS: Mice harboring kinase-inactivating mutations in fps and fer alleles were generated previously. Compound homozygous mice were bred that lack both Fps and Fer kinase activities and progeny were analyzed for potential defects in viability and fertility. Potential differences in hematopoiesis were analyzed by lineage analysis of bone marrow cells, peripheral blood counts, and hematopoietic progenitor cell colony-forming assays. RESULTS: Mice devoid of both Fps and Fer kinase activities were viable and displayed reduced fertility. Circulating levels of neutrophils, erythrocytes, and platelets were elevated in compound mutant mice compared to wild-type controls, suggesting that hematopoiesis is deregulated in the absence of Fps and Fer kinases. Compound mutant mice also showed reduced overall bone marrow cellularity, and lineage analysis revealed elevated CD11b(hi)Ly-6G(lo) myeloid cells, which may reflect increased granulocyte progenitors. Although no differences in the overall number of granulocyte/monocyte colony-forming progenitors were observed, qualitative differences in myeloid colonies from compound mutant mice suggested a role for Fps and Fer kinases in regulating cell-cell adhesion or a skewing in cellularity of colonies. CONCLUSIONS: Mice lacking both Fps and Fer kinase activities develop normally, show reduced fertility, and display defects in hematopoiesis, thus providing evidence for functional redundancy between Fps and Fer kinases in regulating hematopoiesis. PMID- 12901972 TI - Clinical relevance of CD95 (Fas/Apo-1) on T cells of patients with B-cell chronic lymphocytic leukemia. AB - OBJECTIVE: Apoptosis mediated via CD95 (Fas/Apo-1) is a key regulator for the biology of normal and malignant lymphocytes. Although the function of CD95 on B cell chronic lymphocytic leukemia cells (B-CLL cells) has been studied intensively, the clinical importance of CD95 expression on normal T cells in B CLL has not been clarified. This study aimed to investigate whether expression of CD95 on peripheral blood T cells correlates with clinically relevant parameters of B-CLL disease. MATERIALS AND METHODS: Expression of CD95 (Fas/Apo-1) on peripheral blood T lymphocytes of patients with B-CLL was determined using flow cytometry and was correlated with expression of activation markers, sensitivity to apoptosis by anti-CD95, and clinical data, such as blood count, Binet stage, therapy, progression-free probability, and survival probability. RESULTS: Differential CD95 expression did not correlate with activation markers or with levels of apoptosis through anti-CD95. However, high levels of CD95 on T cells from B-CLL patients correlated significantly with low lymphocyte doubling time, increased Binet stages, and requirement for chemotherapeutic treatment. Furthermore, increased cell-surface CD95 on T cells was associated with reduced progression-free probability and poorer survival. CONCLUSIONS: CD95 levels on T cells correlate with the clinical course of B-CLL. Prospective studies appear warranted to investigate whether CD95 on T cells has a direct influence on B-CLL disease progression. PMID- 12901973 TI - Effects of tyrosine kinase inhibitor STI571 on human mast cells bearing wild-type or mutated c-kit. AB - OBJECTIVE: STI571 is a tyrosine kinase inhibitor which inhibits the kinase activity of kit, the receptor for stem cell factor (SCF). Because activating mutations of c-kit affecting codon 816 are associated with human mast cell neoplasms, we determined whether STI571 exerted a similar cytotoxic effect on neoplastic and normal human mast cells. METHODS: We investigated the effect of addition of STI571 in increasing concentrations (0.01 to 10 micromolar) to two HMC-1 human mast cell leukemia cell lines carrying two different activating c-kit mutations in codons 816 or 560, as well as the effect of the drug on short-term bone marrow cultures obtained from patients who carry a mutated codon 816 or wild type c-kit. RESULTS: STI571 failed to inhibit the growth of HMC-1(560,816) cells bearing a codon 816 mutation but effectively suppressed the proliferation of HMC 1(560) carrying c-kit with the wild-type codon 816. STI571 did not induce preferential killing of neoplastic bone marrow mast cells in short-term cultures from patients bearing a codon 816 c-kit mutation. In contrast, STI571 caused a dramatic reduction in mast cells in patients without codon 816 c-kit mutations. CONCLUSION: These results suggest that STI571, while effectively killing mast cells with wild-type c-kit, did not show preferential cytotoxicity to neoplastic human mast cells and thus may not be effective in the treatment of human systemic mastocytosis associated with codon 816 c-kit mutations. PMID- 12901974 TI - Mutant p53 in bone marrow stromal cells increases VEGF expression and supports leukemia cell growth. AB - OBJECTIVE: Genetic alterations, including p53 mutations, have been identified in the stroma of solid tumors and are thought be involved in the induction of tumor growth and metastasis. We tested the hypothesis that somatic molecular alterations in bone marrow stromal cells provide a favorable growth environment for leukemic cells. MATERIALS AND METHODS: We established an in vitro model consisting of stroma expressing mutant p53 (Cys135Ser) to study its ability to support growth of cells from a pre-B acute lymphoblastic leukemia (ALL) cell line. Normal and leukemic bone marrow stromal cells were screened for p53 mutations by mutant-specific ELISA, SSCP, and direct sequencing. Secretion of vascular endothelial growth factor (VEGF) was measured by quantitative ELISA. RESULTS: Transfection of stromal cells with mutant p53 increased synthesis of VEGF and supported the growth of leukemic cells. An ELISA-based assay suggested the occurrence of in vivo p53 alterations in bone marrow stromal cells from 2 of 12 ALL patients screened. Direct sequencing of one of these samples revealed a somatic heterozygous p53 gene mutation (Asp49His). This sample secreted more VEGF and provided increased growth support to leukemic cells. The ability of Asp 49His p53 to increase the expression of VEGF was confirmed with transfection experiments in a p53-null cell line. CONCLUSION: Our findings indicate that genetic alterations, such as p53 mutations, in stromal cells can increase stromal derived support of leukemia growth. Increased synthesis of pro-angiogenic cytokines, such as VEGF, may constitute one possible pathway by which this process is mediated. PMID- 12901975 TI - The emergence of Ph-, trisomy -8+ cells in patients with chronic myeloid leukemia treated with imatinib mesylate. AB - OBJECTIVE: To describe clinical and laboratory features of a cohort of patients with chronic myelogenous leukemia (CML) who developed Ph(-), trisomy 8(+) metaphases while on treatment with imatinib mesylate. PATIENTS AND METHODS: Conventional cytogenetics and triple-color interphase fluorescence in situ hybridization were used to identify 5 of 310 studied patients who, on follow-up analysis, had Ph(-), trisomy 8(+) cells while on therapy. RESULTS: None of the 5 patients had cytogenetic evidence of clonal evolution at the start of treatment with imatinib. All patients developed grade 3 or 4 neutropenia and thrombocytopenia during treatment. The emergence of Ph(-), trisomy 8(+) metaphases was seen at 3, 6, 13, 16, and 18 months from the start of treatment and was present at multiple time points. The maximum number of trisomy 8 metaphases ranged from 25 to 50%. Concomitantly, all patients had a profound suppression of Ph(+) cells (ranging from 0 to 65%) as well as the appearance of normal metaphases, ranging from 6 to 55%. None of the patients has shown clinical or hematologic signs of progression to a more advanced phase of CML. CONCLUSIONS: While on treatment with imatinib mesylate a small group (less than 5%) of patients with CML developed Ph(-) trisomy 8(+) clone associated with pancytopenia. None of the patients developed clinical or hematological signs of progression to a more advanced phase of CML. These observations suggest that identification of trisomy 8 cells may represent clonal Ph(-) cells that were uncovered by treatment with a selective and potent inhibitor of Ph(+) cells. PMID- 12901976 TI - Coexistent naive phenotype and higher cycling rate of cord blood T cells as compared to adult peripheral blood. AB - OBJECTIVE: Umbilical cord blood (UCB) T cells are predominantly CD45RA(+), secrete less cytokines, and have diminished cytotoxicity compared to adult peripheral blood (PB). We hypothesized that the functional impairment of bulk UCB cells results from the relative dominance of immature lymphocyte subsets. In this study we established the physiologic ranges of lymphocyte subsets in UCB, and contrasted those with adult PB. MATERIALS AND METHODS: Four-color FACS was utilized to characterize surface and intracellular protein expression on lymphocyte subsets from fresh unmanipulated UCB and adult PB. RESULTS: We found that UCB contain significantly higher absolute numbers of T cells, NK cells, and B cells than adult PB (p<0.0001). UCB also contains more "naive" cells not only among CD4(+) and CD8(+) T cells but also among B lymphocytes (p=0.003). Most UCB T cells are CD45RA(+)/CD62L(+) "recent thymic emigrants" with smaller TCRgammadelta (p<0.0001) and CD25(+) subsets (p=0.0068). Fewer UCB T cells display HLA-DR and CCR-5 activation markers (p<0.0001) while the CD8(+)/CD57(+)/CD28(-) "suppressor," CD8(+)/CD45RA(+)/CD27(-) "cytotoxic," and skin homing CLA(+) T-cell subsets are absent altogether. Compared with adult PB, more cord blood T cells progress through cell cycle (p<0.0001) and enter apoptosis (p=0.0003). Unlike in adult PB, the majority of proliferating Ki-67(+) T cells in UCB retain a CD45RA(+)/RO(-), CD69(-), CD25(-), HLA-DR(-) "resting" phenotype (p=0.0002). CONCLUSION: Most T and B lymphocytes express a nai;ve phenotype in cord blood while "suppressor" and "cytotoxic" T-cell subsets are absent. Cycling UCB T cells retain a nai;ve immunophenotype that may represent homeostatic expansion rather than antigen-driven proliferation. PMID- 12901977 TI - Telomerized human multipotent mesenchymal cells can differentiate into hematopoietic and cobblestone area-supporting cells. AB - OBJECTIVE: To compare the hematopoietic support provided by telomerized human mesenchymal stem cells (MSCs) and telomerized MSC-derived stromal cells. METHODS: We transfected the human telomerase catalytic subunit (hTERT) gene into primary MSCs to establish hTERT-transduced MSCs (hTERT-MSCs). Stromal induction of hTERT MSCs was performed by replacing the culture medium with Dexter-type culture medium. Hematopoietic support was examined by coculture with cord blood CD34(+) cells. RESULTS: The hTERT-MSCs were morphologically identical with the primary MSCs and expressed surface antigens including CD105, CD73, and CD166. hTERT-MSCs showed a similar doubling time as primary MSCs and continued to proliferate to over 80 population doublings (PD), although the primary MSCs underwent crisis in vitro at 16 PD. The osteogenic, chondrogenic, adipogenic, neurogenic, and stromal differentiation potential of hTERT-MSCs were maintained up to at least 40 PD. The degree of expansion of CD34(+) cells and total number of colony-forming units in culture (CFU-C) upon 12-day coculture with the hTERT-MSC-derived stromal cells were nearly the same as those upon 12-day coculture with hTERT-MSCs (CD34, 33.0 fold+/-2.8-fold vs 36.1-fold+/-1.7-fold of the initial cell number; CFUs, 344.4 fold+/-62.5-fold vs 239.3-fold+/-87.0-fold; CFU-mix, 368.4-fold+/-113.7-fold vs 341.3-fold+/-234.3-fold). However, on day 18 of coculture, the number of cobblestone areas (CA) observed beneath the stromal cells was 15 times higher than that beneath hTERT-MSCs (CA, 146.9+/-54.6 vs 9.4+/-8.1, p<0.01). CONCLUSION: Stromal induction of hTERT-MSCs exclusively enhanced the support of CA formation provided by hTERT-MSCs. Our human hTERT-MSCs will be useful for elucidating the mechanism of the formation of CAs. PMID- 12901978 TI - Adult human bone marrow-derived mesenchymal progenitor cells are capable of adhesion-independent survival and expansion. AB - OVERVIEW: We show the existence of adult human mesenchymal progenitor cells (hMPCs) that can proliferate, in a cytokine-dependent manner, as individual cells in stirred suspension cultures (SSC) while maintaining their ability to form functional differentiated mesenchymal cell types. MATERIALS AND METHODS: Ficolled human bone marrow (BM)-derived cells were grown in SSC (and adherent controls) in the presence and absence of exogenously added cytokines. Phenotypic, gene expression, and functional assays for hematopoietic and nonhematopoietic cell populations were used to kinetically track cell production. Limiting-dilution analysis was used to relate culture-produced cells to input cell populations. RESULTS: Cytokine cocktail influenced total and progenitor cell expansion, as well as the types of cells generated upon plating. Flow cytometric analysis of CD117, CD123, and CD45 expression showed that cytokine supplementation influenced SSC output. The concomitant growth of CD45(+) and CD45(-) cells in the cultures that exhibited the greatest hMPC expansions suggests that the growth of these cells may benefit from interactions with hematopoietic cells. Functional assays demonstrated that the SSC-derived cells (input CFU-O number: 1990+/-377) grown in the presence of SCF+IL-3 resulted, after 21 days, in the generation of a significantly greater number (p<0.05) of bone progenitors (33,700+/-8763 CFU-O) than similarly initiated adherent cultures (214+/-75 CFU-O). RT-PCR analysis confirmed that the SSC-derived cells grown in osteogenic conditions express bone specific genes (Cbfa1/Runx2, bone sialoprotein, and osteocalcin). CONCLUSIONS: Our approach not only provides an alternative strategy to expand adult BM-derived nonhematopoietic progenitor cell numbers in a scalable and controllable bioprocess, but also questions established biological paradigms concerning the properties of connective-tissue stem and progenitor cells. PMID- 12901980 TI - Microtensile bond strength testing of luting cements to prefabricated CAD/CAM ceramic and composite blocks. AB - OBJECTIVES: To investigate the Microtensile bond strength (microTBS) and failure mode of resin cements bonded to composite and ceramic CAD/CAM blocks following various surface treatments. METHODS: Paradigm composite blocks and Cerec Vitablocs received three surface treatments following the control treatment of surface grinding with 600 SiC grit. (1) Application of adhesive resin (Adh), (2) etching with hydrofluoric acid and silanization (HF+S) or (3) combination of the previous two treatments (HF+S+Adh). Three resin cements (Tetric Flow, Nexus 2, RelyX ARC) were applied to these surfaces and built-up in layers. After 24 h water storage at 37 degrees C, the non-trimming version of microTBS test was used to produce 1 mm(2) microbars. The Microbars were subjected to a tensile load using a modified testing device. The broken specimens were examined with a stereomicroscope and SEM to determine the failure mode. RESULTS: All control and adhesive treated groups of the ceramic substrate showed premature debonding during cutting. The overall mean microTBS for the three resin cements bonded to ceramic following HF+S and HF+S+Adh surface treatment, was 27 and 29.2 MPa and for the resin cements bonded to composite substrate was 42.3 and 54.2 MPa, respectively. The mode of failure was 98% adhesive with composite as a substrate and 68% mixed failures with ceramic as a substrate. CLINICAL SIGNIFICANCE: CAD/CAM restorations fabricated from processed composite blocks may have advantage over the ceramic blocks with regard to the higher bond strength with resin cements. PMID- 12901979 TI - Direct visualization of transplanted hematopoietic cell reconstitution in intact mouse organs indicates the presence of a niche. AB - OBJECTIVE: The temporal and spatial behavior of transplanted hematopoietic stem cells (HSCs) within bones remains to be clarified. Our goal is to examine in vivo reconstitution processes and candidate niches in all bones in the mouse body using a new visualization method. MATERIALS AND METHODS: Using bone marrow cells from green fluorescent protein (GFP) transgenic mice, the reconstitution processes of transplanted hematopoietic cells (HCs) under myeloablative or nonmyeloablative conditions were observed sequentially from outside the bones with a fluorescent stereomicroscope. RESULTS: In case of myeloablative transplantation, GFP(+) spots were first detected at the epiphysis of femurs, and in some ribs and vertebrae among all intact bones. Thereafter, engrafted cells proliferated and spread into other bones. In case of nonmyeloablative transplantation with lin(-)Sca-1(+)c-kit(+) cells into W/Wv neonates, characterized by vacant niches because of stem cell defects, GFP(+) cells localized at the epiphysis of femurs and in some vertebrae and ribs, but not in all bones even 4 months after transplantation. CONCLUSION: Our findings show that transplanted HSCs or their immature progenies engraft preferentially at the epiphysis of the femurs or short and flat bones such as ribs and vertebrae. The transplanted cells remain quiescent for at least 4 months under nonmyloablative conditions, which implies the presence of stem cells in a niche. Our approach for the first time graphically demonstrates the kinetics of HCs in vivo and should facilitate analysis of HSC behavior in a three-dimensional mode. PMID- 12901981 TI - Synthesis, characterization and evaluation of urethane derivatives of Bis-GMA. AB - OBJECTIVES: The aims of the study were to synthesize derivatives of Bis-GMA having pendant n-alkyl urethane substituents and to characterize and evaluate their physicochemical properties. METHODS: Stoichiometric amounts of Bis-GMA and n-alkyl isocyanates were reacted in dichloromethane with dibutyltin dilaurate as a catalyst. Volumetric shrinkage, water uptake, degree of vinyl conversion, refractive index and viscosity of resulting urethane monomers and those of Bis GMA were measured. The flexural strengths of their corresponding homopolymers and that of Bis-GMA were also measured. RESULTS: These types of urethane derivatives of Bis-GMA exhibited lower viscosities and were more hydrophobic than Bis-GMA. Generally, the viscosity of these experimental monomers decreased with increasing chain length of the alkyl urethane substituent. Photopolymerization of the new monomers gave high degrees of vinyl conversion compared to Bis-GMA. The experimental monomers also yielded polymers with lower polymerization shrinkages at equivalent degrees of vinyl conversion, than Bis-GMA. The refractive indices of these urethane derivatives were similar to Bis-GMA, but the flexural strengths of their polymers were lower than that of the Bis-GMA homopolymer, decreasing with increasing chain length of the alkyl urethane substituent. SIGNIFICANCE: Because of their excellent overall properties, these new derivatives of Bis-GMA have potential as dental monomers that can improve many properties of resin based dental materials that utilize methacrylate monomer systems. PMID- 12901982 TI - A polymeric device for delivery of anti-microbial and anti-fungal drugs in the oral environment: effect of temperature and medium on the rate of drug release. AB - OBJECTIVES: The use of drug delivery systems in dentistry is a relatively new area of research with the exception of fluoride ion release from polyalkenoate cements and their predecessor silicate cements. The present study is based on the use of a bio-compatible material ethylene vinyl acetate copolymer (EVA) that enables constant release of drugs of therapeutic levels over extended periods of time at doses suitable for the treatment of oral conditions. METHODS: Polymer casting solutions were made by dissolving EVA and the drug in the ratio of 40:1 in 70 ml of dichloromethane at 38 degrees C for 6 h. Thin square films of 3 x 3 cm2 with a thickness of 1 mm were cut from the dry sheet obtained by solvent evaporation technique. Drug loaded samples were extracted for a minimum of 14 days in 10 ml medium (double distilled water or water/ethanol (4:1)) which was replaced daily. Spectral measurements were made to follow changes in optical densities (OD) during release kinetics. Effect of temperature (24 and 37 degrees C) on the rate of drug release was studied and the energies of activation (DeltaE not equal ) were calculated using Arrehenius equation for the diffusion (translocation) of molecules of tetracycline hydrochloride (TTH), doxycycline hydrochloride (DOH), and chlorhexidine diacetate (CDA) in water as extracting medium. Effect of extracting medium (water and water/ethanol (4:1)) was also investigated on the rate of drug release measurements at 24 degrees C. RESULTS: Analysis of variance of the data revealed that significantly enhanced rates were observed at the higher temperature (37 degrees C) and when extracting medium was changed to water/ethanol (4:1) for TTH, DOH and CDA (p<0.0015). The enhanced rate values seem to be due to the formation of channels in the polymer. The largest activation energy (21.83 kcal mol(-1)) observed for CDA was interpreted as due to the highest average molecular weight (626) compared to TTH (481) and DOH (481).Significance. These in vitro rate of drug release measurements will provide a basis for establishing a novel approach (treatment modality) for sustained intra-oral drug delivery over extended time periods using laboratory methods and materials that are readily available to dentists. PMID- 12901983 TI - Polymerization contraction stress in light-cured compomer restorative materials. AB - OBJECTIVE: The magnitude and kinetics of polymerization contraction stress build up may be potential predictors of bond failure of adhesive restorations. The present study determined these properties of seven commercial compomers (Dyract, Dyract AP, F2000 Rasant, Hytac, Compoglass F, Luxat, Glasiosite). METHODS: Polymerization shrinkage was generated by 40 s light curing the test materials (800 mW/cm2). The contraction force induced was recorded for 300 s at room temperature (23-24 degrees C) by means of a Stress-Strain-Analyzer (C factor=0.33). Maximum contraction stress (MPa), coefficient of near linear fit of contraction force/time (gradient) and relative force rate (%/s) of each material were compared with that of two hybrid composites (Tetric Ceram, Prodigy). The statistical analysis was conducted by ANOVA (alpha=0.05) and post hoc Tukey's test. RESULTS: No statistically significant differences in the maximum stress between Glasiosite (2.27+/-0.06 MPa), Hytac (2.31+/-0.07 MPa) and Tetric Ceram (2.21+/-0.11 MPa), and between Compoglass F (2.60+/-0.18 MPa) and Prodigy (2.70+/ 0.06 MPa) were found. The contraction stress of F2000 Rasant (3.41+/-0.09 MPa) and Luxat (3.33+/-0.08 MPa) were significantly highest, whilst Dyract exhibited the significantly lowest shrinkage stress (1.27+/-0.08 MPa) among the tested materials. SIGNIFICANCE: High contraction stress, early start of stress build-up and rapid contraction force development may lead to failure of bond to tooth structure. This study suggested that the contraction stress and kinetic behavior of compomers are generally similar to those of hybrid composites in a dry condition. Dyract might be superior in maintaining the bond with cavity walls compared to conventional hybrid composites in view of its low shrinkage stress. PMID- 12901985 TI - Color stability and hardness in dental composites after accelerated aging. AB - OBJECTIVES: To investigate the color and microhardness changes of five chemically and five light-curing composites as a function of accelerated aging from light exposure. MATERIALS AND METHODS: From each material five composite specimens were embedded in epoxy resin prior to determining the Knoop microhardness of the surface. For analyzing the color DeltaE*=f((L*a*b*)) with a spectrophotometer, three discs per composite were prepared. After measuring the baseline for hardness and color the same specimens were exposed to a xenon arc light and water in a Weather-Ometer machine for a total radiant energy of 150 kJ/m2 and 122 h. The microhardness and the color were again determined following the aging treatment. RESULTS: Each material showed a significant increase in hardness after aging treatment (p<0.05). Comparing the hardness changes (in %) of the light cured materials with the chemically cured materials, no significant difference could be found. Perceptible color differences could be observed for all the materials. Three brands showed small differences with DeltaE*=1.6-2.2, while four composites had DeltaE* ranging from 6.2 to 15.5. A significant correlation between hardness values and color changes could not be established. CLINICAL SIGNIFICANCE: The findings suggest that, since light-curable materials showed significantly more resistance to color changes after accelerated aging by light and water than chemically-cured materials, they may be more esthetically acceptable. Color changes were not correlated with surface hardness changes of the materials after aging. PMID- 12901984 TI - Influence of microstructure and chemistry on the fracture toughness of dental ceramics. AB - OBJECTIVES: the primary aim of this research was to measure fracture toughness for several groups of dental ceramics, and determine how this property is affected by chemistry and microstructure. METHODS: Fracture toughness (KIc) values were obtained using Single Edge Precracked Beam (SEPB) and Single Edge V Notch Beam (SEVNB) methods. Dynamic Young's modulus, which often scales with strength and has been used in explaining the microstructure/toughness relationship on a theoretical basis, was also obtained for the three groups of materials comprising this study. The first group, consisting of micaceous glass ceramics, included model materials that varied systematically in microstructure but not in chemistry. The second group, the feldspathic porcelains, varied significantly in microstructure, but little in chemistry. The ceramics comprising the third group were significantly different in both chemistry and microstructure. RESULTS: Upper toughness limits for the micaceous glass-ceramics and feldspathic porcelains were significantly raised compared to the base glasses, but remained under 2 MPa m(1/2). The highest toughnesses were associated with high percent crystallinity, large grains and high aspect ratios. The third group KIc values were 2.8 MPa m(1/2) for a lithium disilicate glass-ceramic, 3.1 MPa m(1/2) for a glass-infused alumina, and 4.9 MPa m(1/2) for zirconia. SIGNIFICANCE: the correlations between microstructural characteristics and measured properties supports theoretical predictions in the literature. From a practical standpoint, microstructural effects were found to be important, but only within a limited range; the chemistry apparently defined a band of achievable property values. This suggests very large increases in fracture toughness are unlikely to be attained by changes in microstructure alone. A functional relationship determined for the micaceous glass-ceramics enables quantitative predictions of fracture toughness based on the microstructure. PMID- 12901987 TI - Mechanism of strength increase for a hydrothermal porcelain. AB - OBJECTIVES: The objectives of this study were to verify the formation of a hydrolyzed surface layer on Duceram LFC, to determine the effects of such a layer on mechanical material properties, and to identify a specific mechanism responsible for any strength increase observed. METHODS: Specimens were fabricated from dentin porcelain by a vibration blotting technique and were prepared to have either blunt or sharp surface flaws. Half of the specimens underwent accelerated aging. Specimens were fractured in three-point flexure to measure their strength, and fractographic analysis was used to determine fracture toughness and residual surface stress. Surface hardness and elastic modulus were measured using a microindentation method. Porcelain surface topography was examined using atomic force microscopy, and Fourier transform infrared spectroscopy was used to determine the composition of the surface layer. RESULTS: The aging treatment modified the porcelain surface topography but did not create a layer with increased hydroxyl ion content. Porcelain strength increased upon aging, and the increase was proportional to initial flaw severity. The apparent fracture toughness of sharp flaw specimens increased to match that for specimens containing blunt flaws upon aging. Surface hardness and elastic modulus decreased upon aging. SIGNIFICANCE: Previous studies on the strength increase of hydrothermal porcelain were contradictory because a variety of specimen preparation procedures were used. This study resolves the apparent contradiction by determining the effect of specimen preparation on material strength. PMID- 12901986 TI - Strengthening effect of aluminum fluoride added to resin composites based on polyacid-containing polymer. AB - OBJECTIVES: The study investigates the possible strengthening effect of aluminum ions liberated during long-time water storage of experimental resin composites containing a polymer with carboxylic acid groups. METHODS: The organic part of the resin composites was formed by photopolymerization of methacrylate monomers. The monomer mixture was composed of UEDMA, HEMA or TEGDMA, and 0-40 mol% HEMAN (the adduct of HEMA and maleic anhydride). The filler part of the resin composites consisted of a conventional silanated glass filler and 1 or 5 wt% of AlF3 x 3H2O. The rupture strength and bending modulus were measured without water storage and after 1 month, 1 year, and 3 years of water storage at 37 degrees C of the resin composite. RESULTS: The experimental composites that had been stored in water had lower rupture strength and modulus than the composites that were tested without water storage. Specimens containing 30 or 40 mol% HEMAN increased in strength, and specimens containing 20, 30, or 40 mol% HEMAN increased in modulus during the period of water storage. Depending on composition, values for strength and modulus obtained after 3 years of water storage was up to 50% higher than the values obtained after 1 month. SIGNIFICANCE: The strengthening effect of aluminum fluoride may be interpreted as the result of increased crosslinking by aluminum ions and carboxylate groups. The results may serve as a basis for the improvement of the mechanical properties of polyacid-modified resin composites. PMID- 12901988 TI - Measurement of the diffusion of liquids into dental restorative resins by stray field nuclear magnetic resonance imaging (STRAFI). AB - OBJECTIVES: The purpose of this investigation was to determine the diffusion mechanism for water/ethanol mixtures in a diacrylate dental resin by direct observation of the absorbed liquid profiles using NMR microimaging. METHODS: Frequency-swept stray-field magnetic resonance imaging (STRAFI) was used. Solutions containing 25-65% by volume ethanol remained in contact with visible light cured 54% TEGDMA: 46% modified TUDMA sheets while measurements were made. The diffusion profiles were recorded periodically for diffusion times up to 10 h, to a depth of 360 microm and with readings taken at 24 microm intervals. RESULTS: For all liquid mixtures, diffusion was found to be Fickian with coefficients that increased progressively and smoothly with alcohol content in the diffusing mixture, from 2.4 x 10(-13) to 150 x 10(-13) m2 s(-1). A rule of mixtures approach, as suggested by Kwei and Zupko, gave a satisfactory description of the ethanol fraction dependence of the diffusivity. SIGNIFICANCE: Frequency-swept STRAFI offers a new and unique opportunity to produce spatially resolved measurements of the liquids in dental resins to high resolution. In this study, absorption was investigated since an understanding of its mechanism is fundamental to limiting consequent environmental degradation. STRAFI has great potential for other applications, for example drying, liquid exchange, etc. Since STRAFI can discriminate 1H in the liquid from those in softened polymer additional applications are envisaged. PMID- 12901989 TI - Surface analysis of dental amalgams by X-ray photoelectron spectroscopy and X-ray diffraction. AB - OBJECTIVES: It is important to characterize the surface of dental amalgam in order to understand the process of mercury release from amalgam restorations in the oral cavity. The mercury evaporation occurs not only from the newly made restoration but also from the set material. METHODS: The surfaces of four different types of amalgams, which had been well set, were analyzed with X-ray photoelectron spectroscopy (XPS) and X-ray diffraction (XRD) and the relationship between surface compositions and mercury release was studied. Fresh amalgam surfaces as well as aged surfaces, which were stored for 30 days in air, were investigated using XPS and the chemical states of amalgam components and oxygen were studied. The aged surfaces were also characterized with XRD and grazing angle XRD. RESULTS: With increased oxidation, the surface contents of tin and oxygen were increased in all amalgams. In contrast, the surface contents of copper and mercury were decreased. An increase of zinc or indium content were observed in zinc or indium containing amalgams, respectively. A surface layer enriched with indium and oxygen was clearly detected by XPS but not with grazing angle XRD. SIGNIFICANCE: The thickness of the enriched surface layer is estimated to be in the order of few nanometer, which is approximately equal to the analysis depth of XPS. In addition, the presence of metallic elements, like tin and zinc, that readily form a stable oxide layer at the surface suppress the release of mercury. PMID- 12901990 TI - Mechanical property characterization of resin cement after aqueous aging with and without cyclic loading. AB - OBJECTIVE: The purpose of this study was to determine changes in flexural properties of resin cement under cyclic loading and aqueous aging. METHODS: Panavia F flexural modulus and strength were measured by static loading to failure after 48-h, 31-d, and 60-d aqueous aging at 37 degrees C with and without cyclic loading at clinical stress levels. Six specimens were used for each experimental condition. Scanning electron microscopy (SEM) was also used to characterize the morphology of the fractured surfaces to potentially identify resin cement components particularly susceptible to degradation under simulated clinical function. RESULTS: A two-factor ANOVA (p< or =0.05) and Least Significant Difference post hoc test indicated that cyclic loading produced a significant increase in the flexural modulus with no significant effect on the flexural strength. In contrast, aqueous aging time produced a significant decrease in flexural strength with no effect on the flexural modulus. The SEM fracture analysis indicated that resin matrix fracture occurred in static-aqueous specimens; while in the aqueous-cycled specimens, resin matrix fracture occurred in addition to an increasing proportion of filler/resin interface fracture as a result of both increased aqueous aging and increased flexural cycles. SIGNIFICANCE: Evidence suggests that after aqueous aging with cyclic loading to simulate resin cement clinical function, initial degradation may be related to breakdown of the filler/resin interface bond. Such breakdown is potentially reflective of slow crack propagation that may contribute to in vivo resin cement cohesive failure. PMID- 12901991 TI - A palladium-free gallium-based alloy: analysis of composition and microstructure. AB - OBJECTIVES: To study the microstructure of a palladium-free gallium-based alloy (Galloy) before and after mixing to identify the metallic phases produced by the setting reaction and assess the quantitative elemental analysis of the phases present. METHODS: The Galloy powder was analyzed using X-ray diffraction (XRD) and scanning electron microscopy (SEM) and the particle size distribution was determined using a high resolution particle size analyzer. Cylindrical test specimens were mechanically condensed in accordance with ISO 1559: 1986 for XRD, energy dispersive spectroscopy (EDS) and SEM analyses of the set alloy. Disc shaped specimens were prepared by manual condensation to compare the width of the reaction zone with specimens condensed mechanically. RESULTS: The Galloy powder consisted of spherical particles varying between 0.1 and 40 microm in diameter which contained a network of grooves and clefts extending deep within the substance of the particles. The reaction zone appeared 'grainy' and 'fragmented' compared with the well defined reaction zone previously observed for Pd containing gallium-based alloys. The student t-test indicated the width of reaction zone varied significantly (P<0.001) with method of specimen fabrication.Significance. The significantly larger reaction zone in the manually condensed specimens was possibly due to failure of the condensation technique to force out excessive liquid alloy present in the triturated mass. The surface topography of the Pd-free Galloy powder particles could be responsible for the characteristically fragmented and ill-defined reaction zone observed in the set material compared with the well defined reaction zone previously observed for Pd containing gallium-based alloys. PMID- 12901993 TI - Resin-modified glass ionomer cement and self-cured resin composite luted ceramic inlays. A 5-year clinical evaluation. AB - OBJECTIVE: This study evaluated IPS Empress ceramic inlays luted with two chemical-cured luting agents, a resin-modified glass ionomer cement (Fuji Plus (F)) and a resin composite (RC) (Panavia 21 (P)). METHODS: Seventy-nine ceramic inlays were placed in Class II cavities in 29 patients. At least two inlays were placed in each patient to compare the luting techniques intra-individually. In each patient half of the inlays were luted with F and the other half with P. The inlays were evaluated clinically, according to modified USPHS criteria (van Dijken, 1986), at baseline, after 6 months, and yearly during 5 years. RESULTS: At 5 years, 71 inlays were evaluated. Two small partial fractures were observed at 3 years (1P, 1F). One inlay showed recurrent root caries at 4 years (P). Four inlays, two in each group showed non-acceptable color match (2P, 2F). Small defects were observed in 4 inlays (2P, 2F). A slight ditching of the cement margins was observed in both luting groups but did not seem to increase during the second half of the evaluation. No significant difference in durability was observed between the two luting agents. SIGNIFICANCE: IPS Empress inlays luted with the chemical-cured RC and the resin-modified glass ionomer cement functioned satisfactory during the 5 years follow-up. PMID- 12901994 TI - Determination of methyl mercury in dental-unit wastewater. AB - OBJECTIVE: The objective of this investigation was to establish whether monomethyl mercury (MMHg) is present in dental-unit wastewater and if present, to determine the concentration relative to total mercury. METHODS: Wastewater samples were collected over an 18-month period from three locations: at the dental chair; at a 30-chair clinic, and at a 107-chair clinic. Total mercury determinations were completed using United States Environmental Protection Agency's (USEPA) method 1631. MMHg was measured utilizing modified USEPA method 1630. RESULTS: The total mercury levels were found to be: 45182.11 microg/l (n=13, SD=68562.42) for the chair-side samples, 5350.74 microg/l (n=12, SD=2672.94) for samples at the 30-chair clinic, and 13439.13 microg/l (n=13, SD=9898.91) for samples at the107-chair clinic. Monomethyl Hg levels averaged 0.90 microg/l (n=13, SD=0.87) for chair side samples, 8.26 (n=12, SD=7.74) for the 30-chair facility, and 26.77 microg/l (n=13, SD=34.50) for 107-chair facility. By way of comparison, the MMHg levels for the open ocean, lakes and rain are orders of magnitude lower than methyl mercury levels seen in dental wastewater (part per billion levels for dental wastewater samples compared to part per trillion levels for samples from the environment). SIGNIFICANCE: Environmentally important levels of MMHg were found to be present in dental-unit wastewater at concentrations orders of magnitude higher than seen in natural settings. PMID- 12901992 TI - Weibull analysis and flexural strength of hot-pressed core and veneered ceramic structures. AB - OBJECTIVE: To test the hypothesis that the Weibull moduli of single- and multilayer ceramics are controlled primarily by the structural reliability of the core ceramic.Methods. Seven groups of 20 bar specimens (25 x 4 x 1.2 mm) were made from the following materials: (1) IPS Empress--a hot-pressed (HP) leucite based core ceramic; (2) IPS Empress2--a HP lithia-based core ceramic; (3 and 7) Evision--a HP lithia-based core ceramic (ES); (4) IPS Empress2 body--a glass veneer; (5) ES (1.1 mm thick) plus a glaze layer (0.1 mm); and (6) ES (0.8 mm thick) plus veneer (0.3 mm) and glaze (0.1 mm). Each specimen was subjected to four-point flexure loading at a cross-head speed of 0.5 mm/min while immersed in distilled water at 37 degrees C, except for Group 7 that was tested in a dry environment. Failure loads were recorded and the fracture surfaces were examined using SEM. ANOVA and Duncan's multiple range test were used for statistical analysis. RESULTS: No significant differences were found between the mean flexural strength values of Groups 2, 3, 5, and 6 or between Groups 1 and 4 (p>0.05). However, significant differences were found for dry (Group 7) and wet (Groups 1-6) conditions. Glazing had no significant effect on the flexural strength or Weibull modulus. The strength and Weibull modulus of the ES ceramic were similar to those of Groups 5 and 6. SIGNIFICANCE: The structural reliability of veneered core ceramic is controlled primarily by that of the core ceramic. PMID- 12901995 TI - The effect of amalgam bonding on the stiffness of teeth weakened by cavity preparation. AB - OBJECTIVE: The objective of this study is to evaluate the effect of amalgam bonding on the stiffness of teeth weakened by cavity preparation. METHODS: Strain gages were bonded to maxillary premolars. The rigidity was tested by applying a load to a sequence of sound, prepared and restored teeth as follows: sound tooth, MOD preparation, amalgam restoration, amalgam removed recovering the MOD preparation, bonded amalgam restoration, bonded amalgam removed recovering the MOD preparation, bonded composite restoration. The relative stiffness (RS) and relative deformation (RD) of each condition for each cusp to that of the sound tooth was determined. RESULTS: The premolar cusps were deformed 1.80, 2.14, and 2.32 times more than the cusps of the sound tooth for the three succeeding MOD preparations. For these three preparations, the stiffness of the premolar cusps was 0.58, 0.48, and 0.46 relative to a stiffness of 1.00 for the sound tooth. The deformation was 1.77, 1.27, and 1.16 for the non-bonded amalgam, the bonded amalgam, and the bonded composite, respectively, corresponding to a mean RS of 0.59, 0.80, and 0.88. The calculated mean stiffness parameter C (standard deviation) was 2.6% (6.9) for the amalgam restoration, 62.5% (12.8) for the bonded amalgam restoration, and 77.8% (15.8) for the bonded composite restoration. The stiffness parameter C measured the extent to which the procedure returned the stiffness of the restored tooth to the original stiffness of the intact tooth (100%). SIGNIFICANCE: Cavity preparation reduced the stiffness and weakened the tooth. Restoring the prepared tooth with unbonded amalgam did not restore the lost tooth stiffness. Restoring the prepared tooth with bonded amalgam or with bonded composite recovered a significant portion of the lost tooth stiffness. It was concluded that bonding amalgam to tooth structure could partly restore the strength and rigidity lost by the cavity preparation. This might lead to a reduction in cuspal flexure and the incidence of tooth fracture due to fatigue. PMID- 12901996 TI - Stress-relieving and porcelain firing cycle influence on marginal fit of commercially pure titanium and titanium-aluminum-vanadium copings. AB - OBJECTIVES: This study evaluated: (1) the marginal fit of copings fabricated from commercially pure titanium (cp Ti) and titanium-aluminum-vanadium alloy (Ti-6Al 4V) after each firing cycle for a recommended porcelain; and (2) the effectiveness of a stress-relieving treatment on the fit. The results were compared with copings made with Pd-Ag alloy submitted to the recommended porcelain firing protocol. METHODS: Bovine teeth were machined for metal-ceramic crowns and die casts were obtained for waxing patterns. The firing cycle of titanium porcelain was performed on cp Ti and Ti-6Al-4V copings obtained using the Rematitan System. A stress-relieving treatment (cp Ti treated and Ti-6Al-4V treated groups) was performed on original invested castings at the first porcelain firing temperature (800 degrees C). Fit (microm) was measured in the 'as cast' condition and after each porcelain firing cycle with a traveling microscope. The values were compared using the analysis of variance and Tukey's test (P<0.05), the Pd-Ag group was compared to titanium groups using Dunnett's test (P<0.05). RESULTS: The marginal fit values in the as-cast condition (initial) were 84.43 microm for Ti-6Al-4V not treated, 76.35 microm for Ti-6Al-4V treated, 86.09 microm for cp Ti not treated, 97.66 microm for cp Ti treated and 58.11 microm for Pd-Ag group. The Ti-6Al-4V, cp Ti, and Pd-Ag groups exhibit significant differences in marginal fit values when the as-cast condition (initial) was compared with those after the recommended porcelain firing protocol (by Tukey's test at 5% probability level). No significant differences were observed between stress-relieving treated and not treated groups.Significance. The firing cycles used for porcelain build-up may worsen the fit of titanium copings to values that suggest no inferences to the detriment of clinical applications. PMID- 12901997 TI - Torsion of a wandering spleen: acute abdominal presentation. AB - Torsion of a wandering spleen is a rare but fulminant condition and is part of the differential diagnosis in patients presenting with acute abdominal pain. It results due to absence or laxity of the various ligaments supporting the spleen. Patients may be asymptomatic or may present with acute abdominal pain. It may occur in people of all ages, with a predilection for male patients under 10 years of age and for female patients in older age groups, being most common in multiparous women. Early intervention is necessary to reduce the risk of splenic infarction and other complications. An awareness of the condition together with use of appropriate medical imaging can lead to the correct diagnosis. PMID- 12901998 TI - Congenital cleft spleen with CT scan appearance of high-grade splenic laceration after blunt abdominal trauma. AB - The spiral computed tomography (CT) scan has made the diagnosis of traumatic injury increasingly rapid and accurate, especially in cases of solid parenchymal organ injury that follows blunt abdominal trauma. Nonetheless, this valuable method of diagnosis can be confounded when anatomical variances are encountered in the setting of concomitant associated injuries. We present here a case where a congenitally bi-lobed spleen was diagnosed by CT scan as an apparent high-grade splenic laceration, prompting emergent laparotomy. PMID- 12901999 TI - Methemoglobinemia due to ingestion of at most three pills of pyridium in a 2-year old: case report and review. AB - Pyridium (phenazopyridine HCl) is a commonly prescribed medication in the treatment of urinary tract infections and is known to cause methemoglobinemia in excessive doses. We report the case of a 2-year-old child who ingested a maximum of three 200-mg tablets (approximately 50 mg/kg) of pyridium and yet developed cyanosis and methemoglobinemia (29.1%), resulting in methylene blue therapy. We urge physicians to consider a period of observation (4-6 h) or to obtain methemoglobin levels in children who ingest even a small number of pyridium tablets because this can represent a toxic dose in a small child. PMID- 12902000 TI - Fever in children less than 36 months of age--questions and strategies for management in the emergency department. AB - Fever is a common pediatric complaint in the Emergency Department. Emergency Physicians often must be conservative in their management of febrile children, as patient follow-up is not always available. A unified approach for the management of febrile infants will be discussed in this article. PMID- 12902001 TI - Thirteen-week abdominal pregnancy after hysterectomy. AB - A case of a 42-year-old woman who presented 11 weeks after a vaginal hysterectomy with a 13-week abdominal pregnancy is presented. The patient had operative intervention, which was complicated by bleeding and a pulmonary embolus. A brief review of the literature relating to post-hysterectomy pregnancy follows. Physicians are urged to consider this diagnosis in women who have had hysterectomies, even if the hysterectomy was performed years earlier. PMID- 12902003 TI - Primary injury prevention in an urban EMS system. AB - Injury prevention is increasingly recognized as an important part of health care delivery. Emergency Medical Services (EMS) personnel may be called upon to deliver primary injury prevention (PIP) by educating both patients and peers about how to avoid future similar injury. The purpose of this study was to determine EMS provider attitudes toward PIP and knowledge and practice of PIP during day-to-day clinical work. A brief survey was administered to a sample of paramedics assessing their attitudes toward primary injury prevention, how often they practice it, and whether or not they have received any PIP education during their training. One hundred sixty-two paramedics completed surveys. Of those surveyed, 70% believe that PIP should be a core mission of EMS systems, and 82% believe PIP should be implemented at the local or regional level. However, only 33% routinely educate their patients how to modify injury risk behaviors, and only 19% routinely provide instruction about proper use of protective devices. Approximately 63% of our paramedics received any PIP education during their training. A majority of paramedics in our survey believe that PIP should be a routine part of EMS. However, many paramedics have not received any PIP education, and few paramedics practice PIP during their clinical practice. PMID- 12902002 TI - Cardiotoxicity and late onset seizures with citalopram overdose. AB - A 31-year-old man ingested 400 mg of citalopram (Celexa) after an argument with his parents and girlfriend 13 h before presentation. Paramedics witnessed the patient having a generalized clonic seizure. The electrocardiogram (EKG) revealed a wide QRS complex, prolongation of the QTc interval, and left bundle branch pattern. He was treated with sodium bicarbonate with resolution of these changes. The patient was continued on a sodium bicarbonate infusion and demonstrated no further EKG abnormalities. Sodium bicarbonate should be considered as a treatment modality in patients with EKG abnormalities of prolongation of QRS or QTc interval after citalopram overdose. PMID- 12902004 TI - The time first-response fire fighters have to initiate care in a midsize city. AB - The objective of this study was to determine whether simultaneously dispatched first-response firefighters (fire) arrive before transporting EMS providers (ambulance) and the amount of time fire has on scene to initiate care. Fire and ambulance dispatch records were obtained for all 9-1-1 responses from four 1 month intervals. Only incidents to which both agencies had been simultaneously dispatched were included. Response time for each agency was determined by subtracting the time of dispatch from the time of arrival. The difference between fire and ambulance response time was the time fire had to initiate care. Both agencies were simultaneously dispatched to 4752 incidents. Average response time for all incidents was 4.0 +/- 2.6 min for fire and 5.3 +/- 2.0 min for ambulance. Fire had 1.3 +/- 3.2 min on average to initiate care. Fire arrived before ambulance for 69% (3262) of requests and for these calls had 2.8 +/- 1.7 min on average to initiate care. Utilization of densely staged first-response fire apparatus in a midsize city may be appropriate because firefighters frequently arrive before ambulances and may have adequate time to initiate lifesaving interventions. PMID- 12902005 TI - Isolated prehospital hypotension after traumatic injuries: a predictor of mortality? AB - In patients with traumatic injuries, prehospital hypotension that resolves by Emergency Department (ED) arrival is of uncertain significance. We examined the impact of prehospital hypotension (PH) in normotensive ED patients with traumatic injuries on predicting mortality and chest/abdominal operative intervention. A retrospective cohort study was conducted of consecutive patients undergoing helicopter transport to two trauma centers between 1993 and 1997. Outcomes were mortality and chest or abdominal operative intervention. Of 545 scene transports, 55 (10.1%) patients were hypotensive on ED arrival, leaving 490 normotensive ED patients. Of 490 patients, 35 (7%) had PH and 455 (93%) had no PH. Multiple logistic regression showed the PH group to have a relative risk for death of 4.4 (95% CI: 1.2-16.6, p < 0.03) and for chest or abdominal operative intervention of 2.9 (1.1-7.6, p < 0.03). In this study of normotensive trauma center patients, prehospital hypotension was associated with increased risk of mortality and significant chest or abdominal injury. PMID- 12902006 TI - Emergency department ultrasound for hemothorax after blunt traumatic injury. AB - Diagnosing hemothorax after blunt trauma may be aided by emergency department (ED) ultrasound (US). Various prior studies have evaluated ED US using different gold standards. A prospective study of blunt trauma patients who underwent computed tomography (CT) scan of the chest, abdomen, or both, was performed. Before CT scan, an US examination was performed specifically to identify free fluid in the thorax. The CT scan findings were used as the gold standard for validation of US results. From July 1998 to June 1999, 142 of 155 patients who underwent US and CT scan for evaluation of blunt trauma were included in this study. The CT scan identified 16 cases of hemothorax among these patients. ED US resulted in 2 true-positive, 2 false-positive, 14 false-negative, and 124 true negative findings. ED US was 12.5% sensitive and 98.4% specific. ED US did not detect small-volume hemothorax identified by CT scan. Future research should focus on further defining the size of hemothorax appreciable with ED US, with increased attention paid to the type of gold standard implemented for its evaluation. PMID- 12902007 TI - Assessing physostigmine's contraindication in cyclic antidepressant ingestions. AB - Ingestion of cyclic antidepressant medications or prolongation of the electrocardiographic QRS interval are commonly considered as contraindications to the use of physostigmine as an antidote for antimuscarinic toxicity. This dictum seems to stem from a few well-publicized cases in which administration of physostigmine was temporally associated with the development of asystole. Before the report of these cases, physostigmine was more frequently used and had been considered a first-line antidote for both the neurologic and cardiac toxic effects of cyclic antidepressant overdose. This apparent inconsistency, and a resurgence of interest in physostigmine as an antidote, begs the question of the appropriateness of this drug's contraindication in all cyclic antidepressant ingestions. Review of the published clinical and experimental evidence provides little support for the clinical utility of using electrocardiographic criteria or the ingestion of cyclic antidepressants as contraindications to the use of physostigmine. PMID- 12902008 TI - Pediatric altered mental status. PMID- 12902009 TI - Vaginal laceration caused by a wood beetle. PMID- 12902010 TI - Prognostic factors in children with Staphylococcus aureus sepsis. PMID- 12902011 TI - Burns from automobile airbags. PMID- 12902012 TI - International report: current state and development of health insurance and emergency medicine in Germany. The influence of health insurance laws on the practice of emergency medicine in a European country. AB - Germany has a comprehensive health insurance system, with only 0.183% of the population being uninsured. Access to office-based medicine and to hospitals is easy and convenient. Due to enormous financial pressures, Germany is currently decreasing the number of beds in hospitals, introducing the Diagnosis Related Groups (DRG), and restricting accessibility to specialists. In contrast to Anglo American countries, Germany follows the concept of bringing the physician to the patient in the prehospital setting, with Emergency Medical Services (EMS) physicians responding to all Advanced Life Support (ALS) calls. Despite a mature EMS system with sophisticated medical equipment and technology, both in the prehospital and hospital setting, logistical issues such as a single emergency telephone number or multidisciplinary Emergency Departments have yet to be established. Within the hospital, this "Franco-German model" considers Emergency Medicine a practice model that does not merit specialty status. Spending restrictions in the health care system, with less access to hospital beds and office-based physicians, will increase the demand for hospital-based emergency care when patients experience problems accessing the medical system. Currently, the German hospital system is unprepared to care for greater numbers of emergency patients. This may call for changes in the German health care system as well as the medical education system, with the introduction of hospital-based Emergency Medicine as its own specialty, similar to Anglo-American countries. PMID- 12902014 TI - The prevalence of female-to-male intimate partner violence in an urban emergency department. AB - Female-to-male intimate partner violence (IPV) recently has become a recognized health care issue. We screened a heterosexual male Emergency Department population for IPV using the HITS scale, a four-question survey. Two hundred eighty-two men were enrolled in the study. Basic demographics, along with the answers to the HITS scale, were analyzed. Of the men screened, 29.3% had a positive history of IPV. Men who were positive for IPV were more likely to score higher on questions regarding the frequency of verbal aggression than actual or threatened physical violence. This study reinforces the need to screen both genders for IPV in the Emergency Department. PMID- 12902013 TI - Ambulance snatching: how vulnerable are we? AB - Out of concern that ambulances might be targeted for hijack for terrorism purposes, we observed security-related behaviors of a cross-section of ambulance crews and their vehicles in Emergency Department ambulance bays. We sent observers to a convenience sample of trauma and suburban Emergency Department ambulance entrances in several states. We observed 151 total ambulance arrivals. Overall, the average time present was 21.5 min, 23.2% of units were left with the engine running, 26.5% were left open, 90.1% were left unattended, 84.1% were unlocked, and 16.6% had a non-crew visitor in the ambulance bay. Several issues were identified demonstrating potential "attractiveness" to individuals who may wish to disrupt Emergency Medical Services or steal an emergency vehicle. We are concerned that this is the case at the majority of ambulance bays in our country. Emergency services agencies should take steps to train their personnel to secure the ambulance. PMID- 12902015 TI - Injury patterns related to use of less-lethal weapons during a period of civil unrest. AB - This case series documents injury patterns related to beanbag and other less lethal (LL) weapons during a period of civil unrest in a major U.S. city. A retrospective review of injuries related to LL weapons usage presenting to an urban Emergency Department (ED) from April 10, 2001 to April 18, 2001 was performed. Patients under 18 years of age and those without signed consent to treat were excluded. Twenty-seven patients were identified. Two were excluded due to lack of signed consent for treatment. Mean age was 28.4 +/- 11.7 years; 76% of patients were male. Significant morbidity was found in 7 (28%) cases with no fatalities. Three (12%) individuals, with the diagnoses of pulmonary contusion, liver laceration, and Achilles tendon rupture, required admission. Two (8%) individuals had delayed complications of pneumonia and post-concussive syndrome. Reports of injury patterns associated with beanbag munitions in the medical literature are limited. Physicians should be aware of the injuries and delayed sequelae associated with their use. PMID- 12902018 TI - Effects of acute and chronic nicotine on somatodendritic dopamine release of the rat ventral tegmental area: in vivo microdialysis study. AB - The objectives of the present study were to examine the effects of acute and chronic nicotine on dopamine (DA) release in the ventral tegmental area (VTA) of Long-Evans rats using in vivo microdialysis. Systemic application of acute nicotine (0.1-0.3 mg/kg, s.c.) significantly increased (145% of baseline) DA release in the VTA. Chronic exposure to nicotine (0.3 mg/kg, s.c.) for 5 days followed by a challenge dose of nicotine (0.3 mg/kg, s.c.) also produced significant enhancement (136% of baseline) of DA release in the VTA. The results suggest that both acute and chronic nicotine treatment exert stimulatory effects on somatodendritic DA release in the VTA. The enhancement of DA release to subsequent challenge nicotine may be susceptible to mild desensitization. PMID- 12902019 TI - The effect of musical training on music processing: a functional magnetic resonance imaging study in humans. AB - Previous studies have demonstrated changes in neuronal activity in trained musicians relative to controls while performing various music processing tasks. In this study the neural correlates of the effect of music training on two aspects of music processing, melody and harmony, are investigated using functional magnetic resonance imaging (fMRI). Fifteen subjects, seven with continuous musical training from early childhood to adulthood and eight without, underwent a passive fMRI listening paradigm designed to test the effects of melodic and harmonic processing. Melodic processing activated the most anterior part of the superior temporal gyrus for both musicians and non-musicians, while harmonic processing activated different visual association areas for musicians relative to non-musicians. The inferior parietal lobules were recruited only by musicians for both tasks. We conclude that musical training results in the recruitment of different neural networks for these aspects of music processing. PMID- 12902020 TI - Neonatal transection of the corpus callosum affects paw preference lateralization of adult Swiss mice. AB - In the present work, the hypothesis that the ontogenetic development of the corpus callosum (CC) affects the establishment of behavioral lateralization was tested by studying paw preference performance in adult Swiss mice that were subjected to mid-sagittal transection of the CC on the first postnatal day. Magnitude and direction of laterality were evaluated independently. No significant differences between groups were found for the magnitude of paw preference. On the other hand, the transected group presented a significant populational bias favoring the left paw that was not present in the control groups. These results lend support to the hypothesis that the development of the CC plays a role in the establishment of the normal pattern of behavioral lateralization. PMID- 12902021 TI - Quantification of brain atrophy in patients with myotonic dystrophy and proximal myotonic myopathy: a controlled 3-dimensional magnetic resonance imaging study. AB - Myotonic dystrophy (DM1) and proximal myotonic myopathy (PROMM or DM2) are two distinct muscular disorders with multisystemic involvement. Both have previously been reported to be associated with cognitive impairment and white matter lesions detected by cerebral magnetic resonance imaging (MRI). In this study, the extent of brain atrophy was investigated in vivo in ten DM1 and nine PROMM patients in comparison to age-matched healthy controls for each group. The diagnosis was confirmed by DNA analysis of all patients. As a quantitative marker, the ratio of brain parenchymal to intracranial volume, called brain parenchymal fraction (BPF), was calculated from 3-dimensional MRI data using an automated analysis technique. Compared to age-matched healthy controls (mean BPF 0.852 +/- 0.032), the BPF in DM1 patients (0.713 +/- 0.031) was highly significantly decreased (P < 0.001). In contrast, the PROMM patients (mean BPF 0.792 +/- 0.029) showed only slightly decreased BPF values (P < 0.05). BPF was not significantly correlated to any of the clinical or genetic parameters in both diseases (disease duration, motor score, educational level, and number of CTG repeats in the expanded allele). In summary, global brain atrophy was demonstrated to occur in both diseases, but was more severely manifestated in DM1 patients. PMID- 12902022 TI - Role of striatal glutamate in heatstroke-induced damage in streptozotocin-induced diabetic rats. AB - In heatstroke related symptoms in streptozotocin-induced diabetic rats (STZ diabetic rats) exposed to a high temperature (43 degrees C) for 60 min, like that in normal rats, the high colon temperature and the decreases of mean arterial pressure, heart rate and cerebral blood flow were abolished by MK-801, the NMDA receptor antagonist. Also, striatal glutamate was markedly released in both animals by this exposure while the release was higher and the survival time was shorter in STZ-diabetic rats than in normal rats. After insulin treatment, like that by MK-801 pretreatment, the striatal glutamate and the incidence of heatstroke in STZ-diabetic rats were markedly reduced. Therefore, heatstroke was more easily induced via the elevated striatal glutamate in STZ-diabetic rats by high temperature exposure. Insulin treatment seems helpful in the prevention of heatstroke in STZ-diabetic rats. PMID- 12902023 TI - L-glutamate decreases glucose utilization by rat cortical astrocytes. AB - Microenvironmental changes including elevated levels of L-glutamate, ionic homeostasis, acidification, and oxygen level are associated with brain insults. Their effects on energy metabolism were studied in cultured astrocytes. L glutamate caused a decrease in lactate accumulation through the activation of transporter in astrocytes in a concentration- and time-dependent manner. Decreases in glucose uptake, lactate synthesis and accumulation as well as increases in mitochondrial activity indicated a switch of the astrocytic metabolism from glycolytic to oxidative. Environmental changes co-operated with L glutamate to regulate the metabolic strategy, e.g. KCl and oxygen deprivation reversed but acidification exacerbated the L-glutamate-mediated decrease in lactate accumulation. Taken together, during chronic exposure, oxidation of non glucose substrates such as L-glutamate fuels the active transport of L-glutamate into astrocytes. PMID- 12902024 TI - Inhaled nitric oxide induces cerebrovascular effects in anesthetized pigs. AB - Although inhaled nitric oxide (NO(i)) is considered to act selectively on pulmonary vessels, EEG abnormalities and even occasional neurotoxic effects of NO(i) have been proposed. Here, we investigated cerebrovascular effects of increasing concentrations of 5, 10 and 50 ppm NO(i) in seven anesthetized pigs. Cerebral hemodynamics were assessed non-invasively by use of near-infared spectroscopy and indicator dilution techniques. NO(i) increased cerebral blood volume significantly and reversibly. This effect was not attributable to changes of macrohemodynamic parameters or arterial blood gases. Simultaneously, cerebral transit time increased while cerebral blood flow remained unchanged. These data demonstrate a vasodilatory action of NO(i) in the cerebral vasculature, which may occur preferentially in the venous compartment. PMID- 12902025 TI - Braking impulse and muscle activation during unplanned gait termination in human subjects with parkinsonism. AB - Persons with parkinsonism (PD) are known to have difficulty with both rapid force production and declination, and may not be able to adjust force levels during bimanual tasks in the upper extremity. We proposed that these deficits might underlie the problems experienced by those with PD when attempting to rapidly terminate locomotor activity. Rapid gait termination requires concomitant increases in braking impulse and decreases in propulsion impulse. We hypothesized that those with PD would be less able to modulate muscle activity and the associated braking impulse during gait termination. Muscular activity of soleus, tibialis anterior and gluteus medius, and braking impulse were compared between a group of subjects with PD and age and gender matched control subjects. Subjects with PD had similar patterns of muscular activation to control subjects although at dramatically reduced levels of amplitude. Consequently, PD subjects were unable to generate sufficient net braking impulse under time-critical conditions. PMID- 12902026 TI - Role of glutamate receptors and an on-going protein synthesis in the regulation of phosphorylation of Ca2+/calmodulin-dependent protein kinase II in the CA3 hippocampal region in mice administered with kainic acid intracerebroventricularly. AB - In an immunohistochemical study, kainic acid (KA, 0.1 microg) administered intracerebroventricularly (i.c.v.) dramatically increased the expression of Ca2+/calmodulin-dependent protein kinase II (CaMK II) and the phosphorylation of CaMK II (p-CaMK II) in the CA3 hippocampal region of mice. Pre-treatment with cycloheximide (a protein synthesis inhibitor; 200 mg/kg) intraperitoneally prevented the expression of CaMK II and phosphorylation of CaMK II induced by KA. In addition, pre-treatment with MK-801 ((5R,10S)-(+)-5-methyl-10,11-dihydro-5H dibenzo[a,d]cyclohepten-5,10-imine; an NMDA receptor blocker, 1 microg, i.c.v.) or CNQX (6-cyano-7-nitroquinoxaline-2,3-dione; a non-NMDA receptor blocker, 0.5 microg, i.c.v.) attenuated the p-CaMK II, but not CaMK II, expression induced by KA. Our results suggest that KA administered supraspinally induces CaMK II and the phosphorylation of CaMK II expression in the CA3 hippocampal region, for which an on-going protein synthesis is needed. Furthermore, both NMDA and non NMDA receptors appear to be involved in supraspinally administered KA-induced phosphorylation of CaMK II. PMID- 12902027 TI - Differential effects of post-implantation time on potassium- versus D-amphetamine evoked dopamine overflow in the striatum of F344 rats. AB - Effects of post-implantation time on potassium (K+)- versus D-amphetamine (D AMPH)-evoked striatal dopamine (DA) overflow were measured using microdialysis in freely moving young and aged Fischer 344 rats. In one group, samples were collected on the day of probe insertion (Day 1 group). In a second group, samples were collected 24 h after probe insertion (Day 2 group). While analyses revealed no significant differences between the two age groups, the 100 mM K+ stimulus evoked a significantly greater amount of DA overflow in the Day 1 group compared to the Day 2 group. The decrease in 3,4-dihydroxyphenylacetic acid (DOPAC) produced by K+ stimulation was not influenced by post-implantation time. The effect of the 250 microM D-AMPH stimulus on DA overflow did not differ between the Day 1 and Day 2 groups, nor did the decrease in DOPAC that accompanied D-AMPH stimulation. These results support the hypothesis that under some stimulus conditions, post-implantation time is an important variable in microdialysis studies. PMID- 12902028 TI - Aminergic control of high-frequency (approximately 200 Hz) network oscillations in the hippocampus of the behaving rat. AB - Hippocampal high-frequency (200 Hz, 'ripple') oscillations were recorded in the CA1 area of behaving rats. The histamine H1-receptor antagonist pyrilamine facilitated while the H2-antagonist zolantidine (5 mg/kg i.p) transiently decreased ripple occurrence. Thioperamide, an H3 antagonist, had no effect. The 5 HT1A-receptor antagonist WAY100635 (50 microg i.c.v.) reduced the occurrence and the intrinsic frequency of ripples. The 5-HT3-receptor antagonist Y-25130 (i.c.v.) increased the number but reduced the amplitude of ripples. All the treatments affected sharp-waves and ripple oscillations to the same extent. Changes of ripple occurrence were not secondary to alterations of behavior. In the light of these divergent actions via different receptor subtypes the net effect of aminergic innervations will be determined by their state-dependent activities and mutual interactions as well as receptor localizations. PMID- 12902029 TI - Combination of 5 Hz repetitive transcranial magnetic stimulation (rTMS) and tactile coactivation boosts tactile discrimination in humans. AB - A combination of 5 Hz repetitive transcranial magnetic stimulation (rTMS) over the left primary somatosensory cortex together with tactile coactivation applied to the right index-finger representation (coac + rTMS) boosted tactile discrimination ability tested on the right index-finger. Applying coactivation alone caused a 0.25 mm lowering in tactile discrimination thresholds. In contrast, after coac + rTMS we found a significant further improvement of discrimination thresholds in comparison to the coactivation-induced perceptual changes alone. We demonstrate that the individual further improvement after coac + rTMS depended on the effectiveness of the coactivation protocol when applied alone. Subjects, who showed little gain in tactile performance after coactivation alone, showed the largest improvement after coac + rTMS implying that the combined application was selective for poor learners. The selective effects of coac + rTMS are discussed in respect to N-methyl-D-aspartate receptor activation. PMID- 12902030 TI - Excitotoxic lesions of the lateral parabrachial nucleus do not prevent cholecystokinin-induced suppression of milk intake in rats. AB - We examined the effect of lateral (visceral) and medial (gustatory) parabrachial nucleus (PBN) lesions on the suppression of milk intake induced with cholecystokinin (CCK) in 24 h food-deprived rats. Irrespective of CCK dose, each type of PBN lesion significantly elevated the consumption of milk. Neither medial nor lateral PBN lesions had any discernible influence on the anorectic action of CCK, however. In combination with the results from a previous study using the same rats (Brain Res. 894 (2001) 288), the present results support the view that the role of the lateral PBN in CCK-induced anorexia depends on the nature of the food used at test. PMID- 12902031 TI - Frontal lobe networks for effective processing of ambiguously expressed emotions in humans. AB - This study examines the neural substrates involved in the recognition of ambiguous facial expressions using functional magnetic resonance imaging. Subjects performed two tasks, one in which they judged facial expressions and another in which they identified gender. Subtraction between ambiguous expression and clear expression conditions revealed the activation of anterior cingulate (ACC), medial frontal (MeFG) and bilateral inferior frontal gyrus (IFG). Structural equation modeling showed that the functional connectivity between these areas was greater with the ambiguous expressions than with the clear ones. The activation of the ACC, MeFG, and right IFG was greater with ambiguous expressions than with ambiguous gender. These results suggest that the neural network involving these frontal regions plays a crucial role in the processing of the ambiguously expressed facial emotions. PMID- 12902032 TI - Mutations in the hemochromatosis gene (HFE), Parkinson's disease and parkinsonism. AB - Iron overload increases oxidative stress and may lead to neurodegenerative disease like Parkinson's disease (PD). We studied the role of mutations in the hemochromatosis gene HFE in PD and other parkinsonism (non-PD PS) in two population-based series. The first series consisted of 137 patients with PD and 47 with non-PD PS, and the second of 60 patients with PD and 25 with non-PD PS. In the first series, PD patients were significantly more often homozygous for the C282Y mutation than controls (P=0.03). Patients with non-PD PS in both series were more often carriers for the C282Y mutation than controls (P=0.009, P=0.006, respectively). Our data are hampered by small numbers, yet suggest that the C282Y mutation increases the risk of PD and non-PD PS. The rarity of this genotype requires a large series of patients to prove our hypothesis. PMID- 12902033 TI - Taurine regulates corticotropin secretion at the level of the supraoptic nucleus during stress in rats. AB - We studied the consequence of taurine release within the supraoptic nucleus (SON) for the hormonal stress response. Rats were chronically implanted with both microdialysis probes in the SON and jugular venous catheters. Three days later the animals received either Ringer's solution or a specific taurine antagonist via retrodialysis directly into the SON during a 10-min forced swimming session, while simultaneously blood samples were collected. Compared to the Ringer's control, treatment with the taurine antagonist significantly attenuated the increase in plasma corticotropin concentration caused by the stressor exposure (P < 0.05, analysis of variance). This finding supports the hypothesis that the hypothalamic-neurohypophysial system is a potent regulator of the hormonal stress response and suggests an important role for taurine in this context. PMID- 12902034 TI - Acetylcholinesterase activation in organotypic rat hippocampal slice cultures deprived of oxygen and glucose. AB - Acetylcholinesterase (AChE) was analyzed in organotypic hippocampal slice cultures (OHSCs) during recovery from a brief period (20 min) of combined hypoxia and hypoglycemia. Simulated ischemia transiently increased AChE activity in OHSCs in a time-dependent manner reaching a 1.5 fold increase at 6 h post-ischemia. The ischemia-induced AChE increase was totally abolished by incubation with 10 microM dizocilpine (MK-801), a neuroprotective NMDA receptor blocker. PMID- 12902035 TI - Human cytomegalovirus induces apoptosis in promonocyte THP-1 cells but not in promyeloid HL-60 cells. AB - The effect of human cytomegalovirus (HCMV) infection on the viability of the cells in the monocyte/myeloid lineage was investigated. Two cell lines at different stages in the differentiation pathway, the less differentiated promyeloid HL-60 and the more differentiated promonocyte THP-1 cells, were used in this study. While the viability of THP-1 cells was significantly impaired by HCMV infection, the viability of HL-60 cells was not affected. The decrease in the viability of THP-1 cells appears to result from the increase in apoptosis following HCMV infection. Interestingly, HL-60 cells were more sensitive than THP 1 cells to the apoptotic effect of other apoptogenic agents such as ultraviolet irradiation and hydrogen peroxide. When HL-60 cells were induced to differentiate by treating cells with 12-O-tetradecanoyl-phorbol 13-acetate (TPA), HCMV infection induced an increase in apoptosis of the differentiated HL-60 cells by TPA. Therefore, HCMV-induced apoptosis in the cells of the myeloid/monocyte lineage appears to depend on the degree of cell differentiation. PMID- 12902036 TI - Cooperative transformation of murine fibroblast NIH3T3 cells by hepatitis C virus core protein and hepatitis B virus X protein. AB - Co-infection with hepatitis B virus (HBV) and hepatitis C Virus (HCV) is associated with increased frequency in the development of hepatocellular carcinoma (HCC). Here, we demonstrated that HBV X protein (HBx) and HCV core cooperate to transform mouse fibroblast NIH3T3 cells. They additively stimulated cell growth, especially in the absence of serum growth factors. In addition, co expression of HBx and HCV core had additive effects on the induction of anchorage independent cell growth as well as on the secretion of matrix metalloproteases, which may contribute to increased metastatic potential. Furthermore, the cells expressing both viral proteins exhibited higher tumorigenicity, as demonstrated in athymic nude mice. PMID- 12902037 TI - Cell cycle regulation of hepatitis C and encephalomyocarditis virus internal ribosome entry site-mediated translation in human embryonic kidney 293 cells. AB - We have established stably transformed human embryonic kidney cell lines (HEK293) containing bicistronic constructs to study regulation of viral internal ribosome entry site (IRES)-mediated translation in vivo. These cells produce Renilla luciferase (Rluc) in a cap-dependent manner, while Firefly luciferase (Luc) synthesis is mediated by IRES elements. Using these cell lines, we demonstrate here that IRES-mediated translation directed by both hepatitis C (HCV) and encephalomyocarditis (EMCV) virus varies with the cell cycle. Experiments involving arrest of the cell lines at different phases of the cell cycle, release of synchronized cells from cell cycle arrest, as well as direct sorting of the cells based on position in the cell cycle have shown that the activity of the HCV and EMCV IRES elements is lowest during the G2/M phase in HEK293 cells. These results suggest that cellular trans-acting factors either stimulate viral IRES mediated translation during G1 and S phases or repress translation during the G2/M phase in HEK293 cells. PMID- 12902038 TI - High prevalence of an IgG response against murine leukemia virus (MLV) in patients with psoriasis. AB - Increasing evidence suggests that human endogenous retroviruses (HERV) could participate in the pathogenesis of autoimmune diseases such as multiple sclerosis and lupus erythematosus. To assess a possible association of murine leukemia virus (MLV)-like group of HERVs with psoriasis we searched for antibodies against MLV proteins in the sera of patients. We showed that anti-MLV antibodies (total) were detected in both psoriatic and control sera. However, they were detected with a higher frequency in psoriasis when compared with controls (91 vs. 53%, respectively, P=0.001). In addition, the IgG response was dramatically increased in psoriasis (86 vs. 8%, respectively, P<0.0001). This immunoreactivity was observed against the products of both the gag and env genes, and the most antigenic proteins were the gp65-70. Moreover, we observed that anti-p30 MLV antibodies reacted with an epidermal protein with a molecular weight of 50 kDa in protein extracts from both normal and psoriatic skin cultures. These observations suggest that HERVs of the MLV-like group could contribute to the immunopathogenesis of psoriasis. PMID- 12902039 TI - The effects of HHV-8 vMIP-II on SIVmac251 infection and replication competent and incompetent SIVmac239Delta3 vectors. AB - Human herpesvirus type 8 vMIP-II has one of the broadest ranges of chemokine receptor binding and therefore a multiplicity of biologic effects, both immunologic and antiviral. These properties make vMIP-II an attractive effector gene to be expressed from gene therapy vectors. The present studies were concerned with both therapeutic approaches: (1) an anti-simian immunodeficiency virus (SIV) biologic, and (2) an effector gene in SIV-based vectors. Regarding its antiviral properties, vMIP-II expressed from bacteria and SIV-based vectors bound the surface of CEMx174 cells and specifically suppressed SIVmac251 infection. A CCR3 monoclonal antibody partially inhibited vMIP-II binding, suggesting that both SIVmac251 and vMIP-II utilize a similar CCR3-like receptor for CEMx174 cell binding. Replication competent SIV-based vectors containing forward and reverse vMIP-II produced neither identifiable vMIP-II nor virions for the first 21 days. Virus replication occurred after this period. Significant sequence alterations in the forward vMIP-II containing replication competent vector transcripts were responsible for the failure of vMIP-II expression. The genetic basis for the initial failure to replicate virus and its later restoration was not determined but appeared in the II-PIMv containing vectors to coincide with deletions and compensatory rearrangements in nef 3' of the polypurine tract. Cells transfected with SIVmac239Delta3DeltaLTR-vMIP-II vectors expressed biologically active vMIP-II that bound CEMx174 cells and suppressed SIVmac251 infection. These data suggest that replication defective SIV vectors expressing immunobiolgic genes such as vMIP-II may prove useful in gene therapies, particularly in augmenting immune responses in chronically infected individuals. PMID- 12902040 TI - Functionality of the 5'- and intergenic IRES elements of cricket paralysis virus in a range of insect cell lines, and its relationship with viral activities. AB - Cricket paralysis virus (Dicistroviridae: Cripavirus) (CrPV) naturally has a wide range of insect hosts which is reflected in its ability to infect several cultured insect cell lines. The expression of viral gene products is controlled by two kinds of internal ribosome entry site (IRES) elements, 5' and intergenic (IG). Using seven cultured cell lines we tested the functionality of both IRES elements by transfection with bi-cistronic RNA constructs. In six of the seven cell lines, expression initiated from both IRES's was significantly higher than that from a control construct and in five of these six lines the expression from the 5'-IRES was higher than that from the IG-IRES. Permissiveness of each of the cell lines for replication of CrPV was tested by infection with purified virions and transfection with viral RNA. Only three of the cell lines were fully permissive for CrPV replication and no correlation between permissiveness and IRES activity was apparent. These results suggest that while IRES function is required for permissiveness, additional cellular and/or viral factors, involved in processing of viral products, packaging of viral particles and interacting with the cap-dependent translation machinery of host cells, are necessary for CrPV to be able to replicate in any given cell. PMID- 12902041 TI - Lytic infection of pseudorabies virus in the presence of spermine, spermidine, or DFMO. AB - The effects of polyamines (spermine or spermidine) and DFMO (an ornithine decarboxylase inhibitor) on the infection of LM (tk-) cells by pseudorabies virus (PRV) were investigated. Results from radioactive methionine labeling showed that the synthesis of viral proteins was not affected; however, the expression of a distinctive cellular protein ( approximately 27 kDa) was induced after the treatment of spermine or spermidine. Using plaque assay, we found that the plaque formation of PRV was not affected by these three reagents either. Furthermore, the effects of these drugs on the transcription of PRV immediate-early gene (IE) promoter were examined by CAT assay, and results showed weak stimulation of transcription by these drugs. Taken together, our results demonstrated that lytic infection of PRV was not influenced by addition of exogenous polyamines or depletion of endogenous polyamines; this conclusion was similar to earlier studies by using herpes simplex virus. PMID- 12902042 TI - The S7 gene and VP7 protein are highly conserved among temporally and geographically distinct American isolates of epizootic hemorrhagic disease virus. AB - Complete sequences of genome segment 7 (S7) from six isolates of epizootic hemorrhagic disease virus serotype 1 (EHDV-1) and 37 isolates of serotype 2 (EHDV 2) were determined. These isolates were made between 1978 and 2001 from the southeast, mid-Atlantic, Midwest and intermountain United States. Analysis of the S7 sequence similarities showed 98.1% identity among the EHDV-1 isolates and 91.0% identity among the EHDV-2 isolates. Comparison of the deduced amino acid similarities showed an even greater degree of similarity among the isolates (100% among the EHDV-1 isolates and 98.9% identity among the EHDV-2 isolates). There was only 75.8% identity between the EHDV-1 and EHDV-2 isolates at the nucleic acid level; however, there was 93.7% identity between the two groups at the amino acid level. The ratio of non-synonymous to synonymous nucleotide indicates a strong selection for silent substitutions. There was no evidence for reassortment between EHDV-1 and EHDV-2 isolates. The high degree of conservation of S7 gene codons and the VP7 protein, suggests that little variation is allowed in preserving the function of this protein. The high degree of conservation also validates the use of diagnostic tests for EHDV based on S7 and VP7. PMID- 12902044 TI - Central antinociceptive effect of a hydroalcoholic extract of Dioclea grandiflora seeds in rodents. AB - The acute treatment of rats and mice with a hydroalcoholic extract from the seeds of Dioclea grandiflora (EHDg) at doses of 250 and 500 mg/kg, by intraperitoneal or oral administration, produced a significant antinociceptive effect in the tail flick and hot plate tests, an effect which was inhibited by naloxone. EHDg given to mice daily for 30 days at a dose of 500 mg/kg, did not cause any observable toxic effect nor any alteration in the pattern of antinociceptive response by the tail immersion test during the course of this treatment. These results suggest that EHDg has a central antinociceptive action devoid of tolerance effect typical of opioid drugs. PMID- 12902045 TI - Mechanism of anti-nociceptive effects of Asarum sieboldii Miq. radix: potential role of bradykinin, histamine and opioid receptor-mediated pathways. AB - The radix of Asarum sieboldii Miq. (AR) has been used to treat pain and inflammation in Korea. The present study was conducted to gain insights into the mechanism of actions regarding anti-nociceptive and anti-inflammatory activities of AR. Administration of methanol extract of AR caused dramatic anti-nociceptive effects based on acetic acid writhing and tail-flick assay. When naloxone (Nx) was pre-treated, AR extract failed to exert such anti-nociceptive effect in the tail-flick assays. These results suggest that AR extract have opioid-like activity. It also exerted significant anti-inflammatory effects in the rat paw edema assay. AR extract caused inhibition in the bradykinin (BK)/histamine mediated ileum contractions of guinea pig. Taken together, these results provide evidence that the methanol extract of AR exerts anti-nociceptive and anti inflammatory effects by activating opioid receptor as well as by inhibiting bradykinin and histamine-mediated actions. PMID- 12902046 TI - Antinociceptive and anti-inflammatory effects of Croton malambo bark aqueous extract. AB - Croton malambo (K.) bark aqueous extract, popularly known in Venezuela as "palomatias" or "torco" was tested for acute toxicity and for its anti inflammatory and antinociceptive effects using tail flick and writhing syndrome tests models, respectively. Croton malambo aqueous extract (6.15 mg/kg i.p.) administered intraperitoneally had a significant antinociceptive and anti inflammatory effects compared to acetylsalicylic acid (200mg/kg p.o.) and sodium diclofenac (5.64 mg/kg p.o.). Studies to determine correlation between chemical composition and pharmacological activity are underway. PMID- 12902047 TI - Oral subchronic toxicity of aqueous crude extract of Plantago australis leaves. AB - The toxic effects of chronic oral administration (60 days) of aqueous crude extract (AE) of Plantago australis Lam. (Plantaginaceae) in rats at doses of 850 and 1700 mg/kg on biochemical (ALT, AST, creatinine, urea, glucose, alkaline phosphatase, total proteins and albumin), hematological (complete hemogram), and histopathological (heart, lung, liver, kidney, esophagus, stomach and gut) parameters were studied. All biochemical and hematological parameters were found to be in the normal range, but ALT in animals that received AE of 850 mg/kg was higher. Histopathological analysis of organs, especially the liver did not present alterations. PMID- 12902048 TI - Traditional herbal drugs of Bulamogi, Uganda: plants, use and administration. AB - We present here an inventory of the medicinal plants of Bulamogi county in Uganda, including their medicinal use, preparation and administration modes. Fieldwork for this study was conducted between June 2000 and June 2001 using semi structured interviews, questionnaires, and participant observation as well as transect walks in wild herbal plant collection areas. We recorded 229 plant species belonging to 168 genera in 68 families with medicinal properties. A large proportion of these plants are herbaceous. The medicinal plants are mainly collected from the wild. Some species, such as Sarcocephalus latifolius (Smith) Bruce, are believed by the community to be threatened by unsustainable intensities of use and patterns of harvesting. Particularly vulnerable are said to be the woody or the slow growing species. Herbal medicines are prepared as decoctions, infusions, powders, or as ash, and are administered in a variety of ways. Other concoctions consist of juices and saps. The purported therapeutic claims await validation. Validation in our opinion can help to promote confidence among users of traditional medicine, and also to create opportunities for the marketing of herbal medicines and generate incomes for the community. The processing, packaging and storage of herbal medicines is substandard and require improvement. PMID- 12902049 TI - Antidiabetic activity of Terminalia catappa Linn fruits. AB - In view of alleged antidiabetic potential, effect of the petroleum ether, methanol, and aqueous extracts of Terminalia catappa Linn (combretaceae) fruit, on fasting blood sugar levels and serum biochemical analysis in alloxan-induced diabetic rats were investigated. All the three extracts of Terminalia catappa produced a significant antidiabetic activity at dose levels 1/5 of their lethal doses. Concurrent histological studies of the pancreas of these animals showed comparable regeneration by methanolic and aqueous extracts which were earlier, necrosed by alloxan. PMID- 12902050 TI - Free radical scavenging activity of the alcoholic extract of Trewia polycarpa roots in arthritic rats. AB - The alcoholic extract of Trewia polycarpa roots (TPE), which exhibited significant anti-inflammatory activity, was evaluated for the possible mode of action by studying its antioxidant potential in adjuvant-induced arthritic rats. The biological defence system constituting the superoxide dismutase, glutathione peroxidase, ascorbic acid showed a significant increase while the lipid peroxide content was found to decrease to a large extent on TPE treatment thereby indicating the extracts free radical scavenging property. Histopathological studies too supported anti-arthritic potential of the roots of Trewia polycarpa. PMID- 12902051 TI - Effect of the aqueous extract of Chrysanthellum indicum on calcium mobilization and activation of rat portal vein. AB - The effects of the aqueous extract of Chrysanthellum indicum (CI) on calcium activation and mobilization were studied using the rat portal vein. The extract caused a concentration-dependent contraction of the portal vein. KCl (80 mM), norepinephrine (NA, 10(-6)M) and CI (4 mg/ml) evoked sustained contraction of the portal vein. In Ca-free medium (with EGTA) the contractions evoked by these agents were reduced significantly. The times-to-peak of KCl, NA and CI were similar -in normal PSS, but in Ca-free medium the times-to-peak for KCl and CI were greatly increased. The contractions induced by CI were not inhibited by chlorpropamide and prazosin, but were blocked by verapamil. The data obtained suggest that the aqueous extract of CI utilizes extracellular calcium pools to bring about contractile response and this effect might be mediated through the activation of potential-sensitive channels. PMID- 12902052 TI - Neuropharmacological activity of Nigella sativa L. extracts. AB - Pharmacological studies have been conducted on the aqueous and methanol extracts of defatted Nigella sativa L. seeds to evaluate their effects on the central nervous system (CNS) and on analgesic activity. The observations suggest that the two extracts of Nigella sativa possesses a potent CNS and analgesic activity (depressant action especially in the case of the methanolic extract). PMID- 12902053 TI - Effect of ethanolic extract of Khaya senegalensis on some biochemical parameters of rat kidney. AB - The effect of administration of ethanolic extract of Khaya senegalensis (2mg/kg body weight) on some biochemical parameters of rat kidney were investigated. Experimental animals were randomly divided into the control, those administered with the extract for 6 days and those administered with extract for 18 days, respectively. The prolonged administration of the extract resulted in significant reduction in the alkaline phosphatase activities of the kidney and its body weight ratio (P<0.05). In contrast, the same prolonged administration of the extract resulted in significant increase in the serum sodium ion concentration (P<0.05) while there was no significant difference in serum potassium ion concentration when compared to control (P>0.05). Administration of the extract for 6 days produced no significant difference from the control values in all the parameters investigated except in serum urea concentration which produced a significant increase (P<0.05). The available evidence in this study suggest that the ethanolic extract of Khaya senegalensis exerted more deleterious effect on the kidney when administered continuously over a prolonged period than a short one and this will adversely affect the functioning of the kidney. PMID- 12902054 TI - Effect of soluble dietary fibre fraction of Trigonella foenum graecum on glycemic, insulinemic, lipidemic and platelet aggregation status of Type 2 diabetic model rats. AB - The soluble dietary fibre (SDF) fraction of Trigonella foenum graecum (Tf-sdf) has previously been shown to reduce postprandial elevation in blood glucose level of Type 2 model diabetic rats by delaying the digestion of sucrose. The Tf-sdf has now been investigated for its chronic effect on serum fructosamine, insulin and lipid levels, and on platelet aggregation in Type 2 diabetic rats. Tf-sdf was administered orally twice daily at a dose of 0.5 g kg(-1) for 28 days. It lowered the serum fructosamine level (P<0.05) with no significant change in the insulin level as compared with the control. Atherogenic lipids, i.e. triglycerides, cholesterol and LDL-cholesterol were found to decrease significantly in Tf-sdf fed rats (P<0.01). HDL-cholesterol showed an opposite trend (P=0.024), but serum non-esterified fatty acid (NEFA) values paralleled the atherogenic lipids (P=0.001). No significant effect on platelet aggregation (%) was found although there was a tendency to lower the aggregation (P=0.069). It is concluded that Tf sdf has a beneficial effect on dyslipidemia and has a tendency to inhibit platelet aggregation in Type 2 model diabetic rats. PMID- 12902055 TI - Induction of apoptosis by Paljin-Hangahmdan on human leukemia cells. AB - Paljin-Hangahmdan is an Oriental herbal formulation under intensive investigation for its ability to modulate growth and survival in cancer cells. This research was performed to study the anticancer effects of Paljin-Hangahmdan water extract (PHWE) in human promyelocytic leukemia (HL-60) cells. After HL-60 cells were routinely cultured, tetrazolium-based colorimetric assay was performed for cytotoxicity test. Cytotoxicity of PHWE (200-800 microg/ml) in HL-60 cells was increased in a dose- and time-dependent manner. However, PHWE (200-800 microg/ml) had less cytotoxicity in peripheral blood mononuclear cells isolated from a healthy subject. To explore the mechanism of cytotoxicity, we used several measures of apoptosis to determine whether these processes were involved in PHWE induced leukemic cell death. Our results showed PHWE induced the cell shrinkage, cell membrane blebbing, apoptotic body, and DNA fragmentation. In addition, the flow cytometric analysis revealed PHWE (200-800 microg/ml) dose-dependently increased apoptotic cells with hypodiploid DNA contents. These results indicate that PHWE can control leukemic HL-60 cells through apoptosis and may have a possibility of potential anticancer activities. PMID- 12902056 TI - Antipyretic properties of the aqueous and ethanol extracts of the leaves of Ocimum suave and Ocimum lamiifolium in mice. AB - Numerous plant species are used to treat ailments associated with pyrexia in the indigenous health care delivery system of Ethiopia. Notable among these are Ocimum suave and Ocimum lamiifolium. The objective of the present study was thus to evaluate the antipyretic effects of the aqueous and ethanol extracts of the leaves of Ocimum suave and Ocimum lamiifolium in mice. Rectal temperatures were recorded before and after inducing pyrexia as well as after administration of the respective extracts every half an hour for 3h. Parallel experiments were run with a standard antipyretic (acetylsalicylic acid) and the vehicle (distilled water). All the plant extracts showed antipyretic property with reasonable onset and duration of action. Both ethanol and aqueous extracts of Ocimum suave were observed to be more potent than those of Ocimum lamiifolium. Aqueous extract of Ocimum suave and ethanol extract of Ocimum lamiifolium were more potent than their other counterpart extracts. Time dependent antipyretic effect was also observed with some extracts; reduced with time with aqueous extract of Ocimum suave and increased with time with both extracts of Ocimum lamiifolium. PMID- 12902057 TI - Anti-ulcerogenic activity of some plants used as folk remedy in Turkey. AB - Five herbal remedies used as gastroprotective crude drugs in Turkey were assessed for anti-ulcerogenic activity using the EtOH-induced ulcerogenesis model in rat. The crude drugs investigated comprises fruits of Elaeagnus angustifolia L. (Elaeagnaceae), fresh fruits of Hibiscus esculentus L. (Malvaceae), fresh roots of Papaver rhoeas L. (Papaveraceae), leaves of Phlomis grandiflora H.S. Thomson (Lamiaceae) and fresh fruits of Rosa canina L. (Rosaceae). Extracts were prepared according to the traditional indications of use. Under our experimental conditions, all extracts exhibited statistically significant gastroprotective effect with better results for Phlomis grandiflora and Rosa canina (100%). At the concentration under study, both crude drugs were more effective than the reference compound misoprostol at 0.4 mg/kg. Even the weakest anti-ulcerogenic effect observed for Papaver rhoeas roots was found statistically potent (95.6%). Histopathological studies confirmed the results of the in vivo test. PMID- 12902058 TI - Some medicinal plants as immunostimulant for fish. AB - Immunostimulant effects of the dietary intake of various medicinal plant extracts on fish, rainbow trout (Oncorhynchus mykiss), were investigated. For this purpose fish were fed with diets containing aqueous extracts of mistletoe (Viscum album), nettle (Urtica dioica), and ginger (Zingiber officinale). Food containing lyophilized extracts of these plants as 0.1 and 1% was used at a rate of 2% of body weight per day for three weeks. At the end of the experimental period, various parameters of non-specific defence mechanisms, including extracellular and intracellular respiratory burst activities, phagocytosis in blood leukocytes and total plasma protein level were examined. Specific growth rates (SGRs) and condition factors (CFs) of the fish were also measured. Plant materials tested for immunostimulatory food additives caused an enhanced extracellular respiratory burst activity (P<0.001) compared to the control group. Especially the rainbow trout fed with a diet containing 1% aqueous extract of powdered ginger roots for three weeks exhibited a significant non-specific immune response. Phagocytosis and extracellular burst activity of blood leukocytes were significantly higher in this group than those in the control group. All plant extracts added to fish diet increased the total protein level in plasma except 0.1% ginger. The highest level of plasma proteins was observed in the group fed with 1% ginger extract containing feed. PMID- 12902059 TI - Antihyperglycemic effects of three extracts from Momordica charantia. AB - Momordica charantia (L.) (Cucurbitaceae) commonly known as bitter gourd or karela is a medicinal plant, used in Ayurveda for treating various diseases, one of which is diabetes mellitus. In this study, various extract powders of the fresh and dried whole fruits were prepared and their blood glucose lowering effect compared by administrating them orally to diabetic rats. The aqueous extract powder of fresh unripe whole fruits at a dose of 20mg/kg body weight was found to reduce fasting blood glucose by 48%, an effect comparable to that of glibenclamide, a known synthetic drug. This extract was tested for nephrotoxicity, hepatotoxicity and biochemical parameters such as SGOT, SGPT and lipid profile. The extract did not show any signs of nephrotoxicity and hepatotoxicity as judged by histological and biochemical parameters. Thus the aqueous extract powder of Momordica charantia, an edible vegetable, appears to be a safe alternative to reducing blood glucose. PMID- 12902060 TI - Anti-angiogenic, antioxidant and xanthine oxidase inhibition activities of the mushroom Phellinus linteus. AB - Fruiting bodies of Phellinus linteus were extracted with 70% ethanol at room temperature. The Phellinus linteus extract (PL) showed strong anti-angiogenic activity, which was detected using the chick embryo chorioallantoic membrane (CAM) assay. The in vitro antioxidant activities of PL were evaluated using two different bioassays. PL was comparable to Vitamin C in scavenging the stable free radical 1,1-diphenyl-2-picrylhyrazyl (DPPH). It also inhibited lipid peroxidation (LPO) in a concentration-dependent manner. These results suggest that antioxidant and anti-angiogenic activities of Phellinus linteus would be partly responsible for its anti-tumor effect. PMID- 12902061 TI - Pulmonary segmentectomy: results and complications. AB - BACKGROUND: Segmentectomy is an anatomic pulmonary parenchymal-sparing resection performed for certain benign lesions and on selected patients with lung cancer. We reviewed our experience with segmentectomy in this highly select group of patients. METHODS: We retrospectively reviewed the records of 61 patients (5% of all anatomic resections) undergoing 62 segmentectomies from 1991 to 2001. Wedge resections were excluded. The operative indications were suspected or known lung cancer (43), benign disease (12), and metastatic cancer to the lung (7). Median follow-up for patients with malignancy was 28 months (range 1 to 89 months). Actuarial survival was calculated using the Kaplan-Meier method. RESULTS: Segmentectomy was performed in 43 lung cancer patients with pathologic stages of Ia-22, Ib-14, IIa-2, IIb-1, IIIa/IIIb-2, and IV-2. All resection margins were negative for malignancy. Segmentectomy for benign diseases included granulomatous disease (5), pulmonary abscess (2), plasmacytoma (1), and others (4). Complications occurred in 39% (24/62) of patients, including atelectasis requiring bronchoscopy (10/62, 16%), pneumonia (9/62, 14%), air leak more than 7 days (5/62, 8%), and supraventricular dysrhythmias (6/62, 10%). In-hospital mortality was 3% (2 patients). Median length of hospital stay was 6 days (range 4 to 66 days). In the lung cancer patients the locoregional recurrence rate was 0% and the distant recurrence rate was 20%. The 4-year actuarial survival for patients with lung cancer was 72%. CONCLUSIONS: Pulmonary segmentectomy has acceptable morbidity and mortality in selected patients with both benign and malignant lung disease and remains a useful procedure in a thoracic surgeon's armamentarium. Distant, not locoregional recurrence, was responsible for the cancer deaths. PMID- 12902062 TI - Peripheral lung adenocarcinomas: 10 mm or less in diameter. AB - BACKGROUND: Few reports have been published regarding peripheral lung adenocarcinomas that are 10 mm or less in diameter. This is considered to be the smallest tumor size detectable by present diagnostic modalities. METHODS: Clinicopathologic studies were performed in 57 patients with peripheral lung adenocarcinomas of 10 mm or less in diameter. Outcomes were compared with two other groups that consisted of 32 patients with adenocarcinomas between 11 and 15 mm in diameter and 35 patients with adenocarcinomas between 16 and 20 mm in diameter. Tumors were curatively resected between 1992 and 2002. RESULTS: The mean age was 61.7 years. The following three features were more frequent: female sex (78.9%), nonsmokers (77.2%), and cases with carcinoma detected by computed tomography despite negative chest radiography (96.5%). Negative lymphatic invasion (94.7%) was significantly higher. Three cases showed lymphatic invasion that was classified as types E or F, according to Noguchi's classification. There were no cases of lymph node metastasis, pleural involvement, or intrapulmonary metastasis. Well-differentiated type was in 93.0%. Types A and B, which are noninvasive alveolar replacement-type adenocarcinomas, were significantly dominant (86.0%). The 5-year postoperative survival rate was 97.3%, which was significantly better than in the other two groups (75.5%, 78.1%). CONCLUSIONS: Histopathologic features of most peripheral lung adenocarcinomas of 10 mm or less in diameter were types A and B. Types A and B were considered fundamentally indicated for thoracoscopic wedge resections. However, the other types required the standard operation. PMID- 12902063 TI - Minimal alteration of pulmonary function after lobectomy in lung cancer patients with chronic obstructive pulmonary disease. AB - BACKGROUND: The aim of this study was to evaluate the influence of chronic obstructive pulmonary diseases (COPD) on postoperative pulmonary function and to elucidate the factors for decreasing the reduction of pulmonary function after lobectomy. METHODS: We conducted a retrospective chart review of 521 patients who had undergone lobectomy for lung cancer at Chiba University Hospital between 1990 and 2000. Forty-eight patients were categorized as COPD, defined as percentage of predicted forced expiratory volume at 1 second (FEV1) less than or equal to 70% and percentage of FEV1 to forced vital capacity less than or equal to 70%. The remaining 473 patients were categorized as non-COPD. RESULTS: Although all preoperative pulmonary function test data and arterial oxygen tension were significantly lower in the COPD group, postoperative arterial oxygen tension and FEV1 were equivalent between the two groups, and the ratio of actual postoperative to predicted postoperative FEV1 was significantly better in the COPD group (p < 0.001). With multivariable analysis, COPD and pulmonary resection of the lower portion of the lung (lower or middle-lower lobectomies) were identified as independent factors for the minimal deterioration of FEV1. Actual postoperative FEV1 was 15% lower and higher than predicted, respectively, in the non-COPD patients with upper portion lobectomy and the COPD patients with lower portion lobectomy. Finally, we created a new equation for predicting postoperative FEV1, and it produced a higher coefficient of determination (R(2)) than the conventional one. CONCLUSIONS: The postoperative ventilatory function in patients with COPD who had lower or middle-lower lobectomies was better preserved than predicted. PMID- 12902065 TI - The natural history of recurrence after bronchoplastic procedures for non-small cell lung cancer. AB - BACKGROUND: The natural course of recurrence after bronchoplastic procedures for non-small cell lung cancer (NSCLC) has not been described. METHODS: Sex, age, tnm stage, histology, neoadjuvant chemotherapy, disease-free interval (months), exact localisation of tumour recurrence, time between first and second recurrence (months), survival after first and second recurrence (months), causes of death were retrospectively recorded in 83 patients operated between December 1993 and July 2001. RESULTS: One patient was lost to follow-up, five resections were nonradical. Survivors' follow-up lasted 5 to 100.7 months (mean 43.3). Fourteen patients (14.4%) died tumor free. Eleven (13.2%) distant recurrences were diagnosed 1 to 42 months (mean 10.6) postoperatively, eight (9.6%) died 0 to 17 months (mean 7.55) after diagnosis. Nine local recurrences (10.8%)-5 unifocal, 4 multifocal-occurred 2 to 35 months (mean 17.3) postoperatively, eight died 0 to 8 months (mean 2.13) after diagnosis. Nine mixed recurrences (10.8%)-1 synchronous, 8 metachronous-were found (14.8%) 2 to 21 months postoperatively (mean 8.3). All died 4 to 41 months (mean 17.83) after diagnosis. Fourteen mediastinal lymph node recurrences occurred, ten as a primary recurrence and four as secondary. Lymph nodes were involved in all multifocal recurrences. Intrabronchial recurrence was observed in five patients and was always a result of progressive regional lymph node recurrence. CONCLUSIONS: The pattern and natural history of recurrence cannot be sufficiently explained by stage and surgical radicality and suggest different genetic characteristics of the primary tumor. In case of reoperation due to intrabronchial recurrence adjuvant mediastinal irradiation should be considered. PMID- 12902066 TI - Photodynamic therapy for endobronchial metastases from nonbronchogenic primaries. AB - BACKGROUND: Endobronchial metastases (EBM) occur in 2% of nonbronchogenic malignancies and frequently present with hemoptysis or dyspnea. This report summarizes our recent experience with photodynamic therapy (PDT) for EBM. METHODS: All patients who have undergone PDT for the treatment of symptomatic EBM from nonbronchogenic primaries were identified in the Division of Thoracic Surgery database to determine number of treatments, frequency of symptom relief, and patient survival. RESULTS: Endobronchial PDT was administered to 27 patients from April 1997 through December 2000. The predominant primary tumor was renal cell (44%). Endobronchial metastases presented metachronously in 74% of patients. Patients underwent a median of two PDT treatments. Local anesthesia with sedation was used in 76% of treatments. Four patients (15%) developed immediate postoperative respiratory distress. Follow-up bronchoscopy 24 to 48 hours after initial treatment demonstrated tumor necrosis in all cases. Acute relief of hemoptysis and dyspnea was achieved in 85% of patients. Seven patients died within 6 weeks of treatment. Median survival time after PDT was 4 months (range 0.2 to 30 months). CONCLUSIONS: Patients with EBM treated with PDT had a median survival of 4 months and patients with limited metastatic disease survived up to 30 months. Photodynamic therapy was effective in palliating hemoptysis or dyspnea from EBM with an acceptable morbidity, although the 30-day mortality was 22%. PMID- 12902068 TI - Predicted versus observed maximum oxygen consumption early after lung resection. AB - BACKGROUND: The objective of this study was to identify the predictors of underestimation and overestimation of postoperative maximum oxygen consumption (VO(2)max). METHODS: A prospective analysis was performed on 229 patients who had 38 pneumonectomies, 171 lobectomies, and 20 segmentectomies. All patients performed a preoperative and postoperative (on average 9.2 days after surgery) maximal stair-climbing test. Predicted postoperative VO(2)max (ppoVO(2)max) was calculated on the basis of the number of functioning segments removed during operation. The patients were divided into three groups: group A (158 cases), patients with a ppoVO(2)max within 1 standard deviation of the observed postoperative VO(2)max; group B (56 cases), patients with a difference between the observed postoperative VO(2)max and ppoVO(2)max greater than 1 standard deviation (underestimation); and group C (15 cases), patients with a difference between ppoVO(2)max and the observed postoperative VO(2)max greater than 1 standard deviation (overestimation). Univariate and multivariate analyses were performed. RESULTS: The only significant predictor of underestimation was a high percentage of functional parenchyma removed during operation (p < 0.0001). The significant predictors of overestimation were a low percentage of functional parenchyma removed during operation (p = 0.01) and a high preoperative VO(2)max (p = 0.002). CONCLUSIONS: The prediction of postoperative VO(2)max was not accurate in all patients. Those with a large amount of functional lung tissue removed during operation tended to have a postoperative VO(2)max greater than expected. Conversely, those patients with a small amount of functional lung tissue resected tended to have a postoperative VO(2)max lower than predicted. PMID- 12902069 TI - High dose rate brachytherapy in the management of lung transplant airway stenosis. AB - BACKGROUND: Airway complications after lung transplantation remain a major cause of postoperative morbidity and mortality. Interventional bronchoscopic management continues to be the main modality in the management of these problems. METHODS: Four patients with airway stenoses after lung transplantation received high dose rate brachytherapy for management of recurrent stenosis. All 4 patients had been treated with various bronchoscopic interventions, including dilation and stenting, electrocautery ablation, and neodymium:yttrium-aluminum-garnet laser therapy. High dose rate endobronchial brachytherapy was subsequently used in all 4 patients for management of recurrent airway obstruction. The radiation dose for all 4 patients was 3 Gy at a distance of 1 cm from the center of the catheter. RESULTS: All four patients have had routine follow-up after endobronchial brachytherapy treatments. Of the 4 patients, 2 treated with this modality showed a significant response to therapy in that the bronchus remained free of obstruction after treatment; 1 patient had partial improvement, and 1 patient failed to show significant improvement and expired from the sequelae of persistent airway obstruction. CONCLUSIONS: Endobronchial brachytherapy can be an effective modality for managing recurrent stenoses caused by hyperplastic granulation tissue at the bronchial anastomosis. The optimal timing and ideal candidate for intraluminal radiation therapy for this problem remains a challenge and warrants further investigation. PMID- 12902070 TI - Warm ischemia lung protection with pinacidil: an ATP regulated potassium channel opener. AB - BACKGROUND: Ischemia/reperfusion injury remains a limiting factor in lung transplantation. Traditional hyperkalemic preservation solutions are associated with a host of metabolic derangements. ATP-regulated potassium channel openers (PCOs) may provide an attractive alternative to traditional solutions by utilizing inherent mechanisms of ischemic preconditioning. The purpose of this study was to assess warm ischemia graft protection with pinacidil, a nonspecific PCO. METHODS: An isolated recirculating blood perfused ventilated rabbit lung model was used (n = 15). No ischemia control lungs underwent immediate reperfusion (n = 5). Warm ischemia control lungs were flushed with lactated Ringers (LR), stored at 37 degrees C for 2.5 hours and then reperfused for 2 hours (n = 5). PCO protected lungs were flushed with LR + 100 micromol/L pinacidil, stored, and then reperfused (n = 5). Intermittent blood gases were taken from the pulmonary artery and left atria. Every 30 minutes, graft function was assessed with a 10-minute 100% fractional inspired oxygen concentration challenge to measure maximal gas exchange. Lung samples were graded for histologic injury and assayed for myeloperoxidase activity. RESULTS: A mixed models repeated measures ANOVA demonstrated a significant difference between groups. Tukey's honestly significant difference multiple comparison test demonstrated significantly improved graft function and reduced histologic injury with pinacidil protection compared with the warm ischemia controls. There was no significant difference in graft function or pathology grade between the pinacidil protected lungs and the no ischemia controls. A similar trend, although not significant, was seen in myeloperoxdiase activity. CONCLUSIONS: Potassium channel openers with pinacidil can provide pulmonary protection against warm ischemia reperfusion injury. PMID- 12902071 TI - Thirty-year experience with repair of pectus deformities in adults. AB - BACKGROUND: A plethora of studies have described repair of pectus deformities in children, but only few reports have described this repair in adults. The purpose of this study was to review our 30-year experience with surgical repair of pectus deformities in adults. METHODS: A retrospective review of all adult patients (> 16 years old) who underwent repair of congenital pectus deformities from 1971 through 2001. RESULTS: There were 77 patients, 64 men and 13 women. Sixty-eight patients underwent surgery for pectus excavatum and 9 for pectus carinatum; median age was 22 years old (range, 16 to 68 years old). Indication for repair was medical concerns in all patients. Preoperative symptoms were dyspnea on exertion in 43 patients, shortness of breath at rest in 22 patients, chest pain in 8 patients, and palpitations in 8 patients. Preoperative electrocardiogram findings included right bundle branch block in 9 patients, sinus bradycardia in 8 patients, left atrial enlargement in 6 patients, and right atrial dilatation in 5 patients. Patterns of the pectus defect were symmetric and localized in 29 patients, symmetric and diffuse in 21, asymmetric and localized in 18, and asymmetric and diffuse in 9 patients. Intraoperative classifications were severe in 38 patients, moderate in 33 patients, and mild in 6 patients. There were no operative deaths. Complications occurred in 11 patients (14.3%). Mean hospital stay was 4 days (range, 2 to 8 days). Mean follow-up was 12 +/- 7 years (range, 4 months old to 24 years old); 1 patient (1.3%) required reoperation for recurrent pectus excavatum. Patient satisfaction and relief of medical symptoms was excellent in 70 patients (90.9%), good in 6 patients, and fair in 1 patient. CONCLUSIONS: Repair of congenital defects of the sternum in adults can be performed safely with low morbidity and no mortality. Long-term results are excellent with requirement for reoperation rare. PMID- 12902072 TI - Safety and efficacy of the supraclavicular approach to thoracic outlet decompression. AB - BACKGROUND: Thoracic outlet syndrome (TOS) is a clinical diagnosis encountered by both thoracic and vascular surgeons. The goal of surgical therapy involves relieving compression of the neurovascular structures at the superior thoracic aperture. The traditional approach to thoracic outlet decompression has been transaxillary; however more centers are moving toward a more tailored approach through a supraclavicular incision. METHODS: The medical records of 67 patients who underwent surgical decompression between 1993 and 2001 for TOS were retrospectively reviewed. Patient demographics and early outcome were assessed through clinic follow-up. RESULTS: Seventy-two thoracic outlet decompressions were performed on 67 patients with the diagnosis of TOS. Five patients underwent bilateral thoracic outlet decompression. All operations in this time period were safely accomplished through a supraclavicular approach. The syndromes associated with thoracic outlet compression were neurogenic (n = 59), venous (n = 10), and arterial (n = 3). Forty-six of 72 (63.9%) operations resulted in complete resolution of symptoms, 17 cases (23.6%) had partial resolution, and 9 patients (12.5%) had no resolution. There were no deaths and morbidity was minimal with 6 complications (8.3%). CONCLUSIONS: The supraclavicular approach is a safe and effective technique in managing all forms of thoracic outlet compression. PMID- 12902073 TI - Twenty-six years of experience with the modified eloesser flap. AB - BACKGROUND: Empyema thoracis is a common thoracic problem with a multitude of therapeutic options. The modified Eloesser flap (MEF) is one means of dealing with this problem in selected complicated patients. The purpose of this study is to report our 26-year experience with the MEF. METHODS: A review of 78 patients who had a MEF from 1975 to 2001 was performed. RESULTS: There were 52 males (67%) and 26 females (33%). Mean age was 59 +/- 14 years. The overall length of stay was 26 +/- 27 days, while mean postoperative length of stay was 16 +/- 17 days. Microbiology of the empyema cavity revealed a predominance of gram-positive organisms. Before a modified Eloesser flap, all patients failed initial conservative interventions and 23 patients (29%) failed surgical interventions. Operative indications were as follows: parapneumonic effusions, 35 patients (45%); postresectional, 23 patients (29%); tuberculosis related, 7 patients (9%); malignant effusion, 4 patients (5%); esophageal fistulas, 4 patients (5%); abdominal sepsis, 3 patients (4%); and hemothorax secondary to trauma, 2 patients (3%). The inverted-U incision was performed in all patients. Average rib resection was 3 +/- 1 ribs. There were no intraoperative complications and adequate drainage was achieved in all patients. Thirty-day morbidity/mortality was 4 patients (5%): 3 died of sepsis and 1 died of metabolic encephalopathy; although long-term follow-up (mean: 109 +/- 141 months) revealed no additional morbidity related to the MEF. CONCLUSIONS: We demonstrate that MEF can be performed as a safe, definitive surgical procedure for the treatment of chronic empyema thoracis. The MEF remains an important option in the surgical treatment of chronic, complicated empyema thoracis. PMID- 12902074 TI - Intracostal sutures decrease the pain of thoracotomy. AB - BACKGROUND: General thoracic surgeons spend much time dealing and treating patients' pain after thoracotomy. METHODS: Two hundred eighty consecutive patients underwent elective thoracotomy for pulmonary resection. Patients with a history of chronic pain were excluded. One general thoracic surgeon performed all procedures. All patients had a functioning preoperative epidural, a skin incision the width of their latissimus dorsi muscle which was cut, sparing of the serratus anterior muscle, undercutting of the rib, preemptive analgesia of the intercostal nerve before rib spreading, and similar number of chest tubes and postoperative pain management. The first 140 patients had their chests closed with pericostal sutures (stitches placed on top of the fifth rib and on top of the seventh rib), and the next 140 patients had their chest closed with intracostal sutures (stitches placed on top of the fifth rib and through the small holes drilled in the bed of the sixth rib). Pain was objectified by a numeric pain score and by the McGill pain questionnaire at 2 weeks, and 1, 2, and 3 months postoperatively. RESULTS: There were 140 patients in each group, and the groups were matched for age, gender, race, types of pulmonary resections, number of chest tubes, number of broken ribs, length of chest tube duration, and length of hospital stay (p > 0.05 for all). The mean pain score for the pericostal group (P group) at 2 weeks, 1 month, 2 months, and 3 months postoperatively was 5.5, 3.8, 2.3, and 1.6, respectively. For the intracostal group it was 3.3, 1.7, 1.1, and 0.6, respectively (p = 0.004, p = 0.0001, p < 0.0001, and p < 0.0001, respectively). Descriptors of pain in the P group were more likely to be, hot/burning, shooting or stabbing (p < 0.003). CONCLUSIONS: Intracostal sutures seem to be less painful than pericostal sutures at 2 weeks, 1 month, 2 months, and 3 months after thoracotomy. The pain is less likely to be described as burning or shooting. PMID- 12902075 TI - Robotic-enhanced biventricular resynchronization: an alternative to endovenous cardiac resynchronization therapy in chronic heart failure. AB - BACKGROUND: Cardiac resynchronization therapy (CRT) by pacing the left and right ventricles is an emerging option for treatment of severe heart failure with ventricular conduction disturbances. Stimulation through a coronary vein is currently the technique of choice to achieve left ventricular (LV) pacing. Unfortunately, this approach carries significant limitations and drawbacks. Therefore we explored robotic-enhanced thoracoscopic implantation of an epicardial lead as an alternative technique to stimulate the LV in cardiac resynchronization therapy. METHODS: A total of 15 patients were included in this study. Right (atrial and ventricular) leads were implanted classically through the left subclavian vein. Robotic-enhanced thoracoscopy was then performed to implant the LV epicardial lead. RESULTS: Of the 15 patients, 13 underwent successful endoscopic robotic cardiac resynchronization therapy. Two patients underwent conversion to a small thoracotomy. No perioperative complication occurred in the patients who did not undergo conversion. Acute and chronic LV lead thresholds were satisfactory in all patients, improving over time. All were subjectively and objectively improved at 4 months. As compared with conventional methods, the procedural cost was not significantly affected. CONCLUSIONS: Based on this feasibility study, we believe that robotic LV epicardial lead implantation is a valuable option to achieve biventricular resynchronization therapy. It allows for more reproducible acute thresholds for LV pacing and sensing than does the percutaneous approach; enables fine tuning of the LV lead position, thus potentially providing optimal hemodynamic benefit; and avoids the pitfalls and limitations of the endovenous approach. Therefore it deserves further prospective studies to assess its place in the therapeutic armamentarium against heart failure. PMID- 12902077 TI - Arterial grafting results in reduced operative mortality and enhanced long-term quality of life in octogenarians. AB - BACKGROUND: Despite well-established benefits of arterial (ART) grafting, surgeons have been reluctant to use this conduit in octogenarians. This study explores the influence of arterial revascularization on operative and long-term outcomes of coronary artery bypass grafting surgery. METHODS: A retrospective analysis was conducted of 987 consecutive patients 80 years of age or older who underwent isolated coronary artery bypass grafting between January 1989 and November 2000. Patients with saphenous vein graft only (SVG; n = 574) were compared with those receiving arterial and saphenous vein grafts (ART+SVG; n = 413). Mean follow-up for SVG patients was 3.8 years (range, 4 months to 12.6 years) and 98.6% complete, and mean follow-up was 3.1 years for ART+SVG patients (range, 2 months to 11.2 years) and 97.3% complete. RESULTS: Patients with SVG had a significantly higher (p = 0.009) operative mortality (11.1% versus 6.3%) and significantly longer postoperative length of stay (12.9 versus 10.7 days; p = 0.002) than ART+SVG recipients. More ART+SVG than SVG patients were free of all postoperative complications (290 of 413; 70.2% versus 372 of 574; 64.8%; p = 0.086). Multivariable analysis identified SVG as an independent predictor of operative mortality (p = 0.014) and late mortality (p = 0.040). When patients were matched by equivalent propensity scores to receive SVG only, operative mortality was higher for SVG patients in four of the five quintiles. At 10 years, 97.0% +/- 1.2% of SVG and 92.9% +/- 3.7% of ART+SVG current survivors were free of all late major adverse cardiac events (p = 0.565), and 95.5% of SVG patients and 97.5% of ART+SVG patients were in Canadian class 1 or 2 (p = 0.162). On the SF-36 quality-of-life assessment, ART+SVG patients scored significantly higher than both SVG patients and age-adjusted normal subjects. Physical health summary component scores were 36.8 +/- 11.0 for SVG and 41.0 +/- 10.3 for ART+SVG (p = 0.001). Mental health summary scores were comparable for the two groups. CONCLUSIONS: Arterial grafting confers an operative survival benefit, and an enhanced long-term quality of life in elderly patients. PMID- 12902078 TI - Intra- and postoperative predictors of stroke after coronary artery bypass grafting. AB - BACKGROUND: Stroke is a devastating complication of coronary artery bypass graft surgery. An individual's risk of stroke is based in part on preoperative characteristics but also on intra- and postoperative factors. We developed a risk prediction model for stroke based on factors in intra- and postoperative care, after adjusting for a patient's preoperative risk. METHODS: We conducted a regional prospective study of 11,825 consecutive patients undergoing coronary artery bypass graft surgery surgery from 1996 to 2001. Data were collected on patient and disease characteristics, intra- and postoperative care and course, and outcomes. Stroke was defined as "a new focal neurologic deficit which appears and is still at least partially evident more than 24 hours after its onset." Logistic regression identified significant predictors of stroke. RESULTS: The incidence of stroke was 1.5%. The regression model significantly predicted the occurrence of stroke. As compared with cardiopulmonary bypass for less than 90 minutes, cardiopulmonary bypass for 90 to 113 minutes, odds ratio = 1.59, p = 0.022), cardiopulmonary bypass for 114 minutes or more (odds ratio = 2.36, p < 0.001), atrial fibrillation (odds ratio = 1.82, p < 0.001), and prolonged inotrope use (odds ratio = 2.59, p = 0.001) significantly improved our ability to predict stroke. Nearly 75% of all strokes occurred among the 90% of patients at low or medium preoperative risk. CONCLUSIONS: The inclusion of factors associated with intra- and postoperative care and course significantly improved the prediction model. Most strokes occurred among patients at low or medium preoperative risk, suggesting that many of these strokes may be preventable. Reduction in stroke risk may require modifications in intra- and postoperative care and course. PMID- 12902080 TI - Development and validation of a prediction model for strokes after coronary artery bypass grafting. AB - BACKGROUND: A prospective study of patients undergoing coronary artery bypass graft surgery (CABG) was conducted to identify patient and disease factors related to the development of a perioperative stroke. A preoperative risk prediction model was developed and validated based on regionally collected data. METHODS: We performed a regional observational study of 33,062 consecutive patients undergoing isolated CABG surgery in northern New England between 1992 and 2001. The regional stroke rate was 1.61% (532 strokes). We developed a preoperative stroke risk prediction model using logistic regression analysis, and validated the model using bootstrap resampling techniques. We assessed the model's fit, discrimination, and stability. RESULTS: The final regression model included the following variables: age, gender, presence of diabetes, presence of vascular disease, renal failure or creatinine greater than or equal to 2 mg/dL, ejection fraction less than 40%, and urgent or emergency. The model significantly predicted (chi(2) [14 d.f.] = 258.72, p < 0.0001) the occurrence of stroke. The correlation between the observed and expected strokes was 0.99. The risk prediction model discriminated well, with an area under the relative operating characteristic curve of 0.70 (95% CI, 0.67 to 0.72). In addition, the model had acceptable internal validity and stability as seen by bootstrap techniques. CONCLUSIONS: We developed a robust risk prediction model for stroke using seven readily obtainable preoperative variables. The risk prediction model performs well, and enables a clinician to estimate rapidly and accurately a CABG patient's preoperative risk of stroke. PMID- 12902081 TI - Is the use of cardiopulmonary bypass for multivessel coronary artery bypass surgery an independent predictor of operative mortality in patients with ischemic left ventricular dysfunction? AB - BACKGROUND: l Coronary artery bypass grafting for patients with ischemic left ventricular dysfunction (ILVD) remains superior to medical therapy in terms of long-term survival. Recently, off-pump coronary artery bypass surgery has been shown to be very promising in achieving functional improvements with favorable operative mortality in this challenging group of patients. The aim of this study was to assess the risk factors responsible for operative mortality in this group of patients. METHODS: The records of 305 consecutive ILVD patients, who underwent primary isolated coronary artery bypass grafting for multivessel disease at The National Heart and Lung Institute, Imperial College, University of London, between January 1999 and January 2002, were reviewed retrospectively. Patients were considered to have ILVD if they had a left ventricular ejection fraction of 0.30 or less on preoperative coronary angiography. One hundred six patients were operated on using the off-pump coronary artery bypass surgery technique, and 199 patients were operated on using the conventional coronary artery bypass grafting technique with cardiopulmonary bypass. RESU;TS: Seven (6.6%) patients died in the off-pump coronary artery bypass surgery group, whereas 28 (14.1%) patients died in the cardiopulmonary bypass group (p = 0.05). Univariate analysis of all the preoperative characteristics was performed to identify the potential predictors of mortality in the whole group of ILVD patients. Potential predictors of mortality included symptom status (stable/unstable), chronic obstructive airway disease, dyspnea grade III and IV on the New York Heart Association classification, intravenous nitrates, preoperative use of intraaortic balloon pump, ventricular tachycardia or ventricular fibrillation, body surface area less than 2, and cardiopulmonary bypass. Only ventricular tachycardia or ventricular fibrillation was proved to act as an independent predictor of operative mortality in this group of ILVD patients, with an odds ratio of 29.6 (95% confidence interval, 8.9 to 98). CONCLUSIONS: This study showed that using cardiopulmonary bypass for multivessel coronary artery bypass grafting in patients with ILVD was not proved to act as an independent predictor of operative mortality. PMID- 12902085 TI - Phrenic nerve injury associated with high free right internal mammary artery harvesting. AB - BACKGROUND: The right phrenic nerve is at risk of injury during high mobilization of the right internal mammary artery (RIMA). The incidence and implications of this injury have not been previously defined. METHODS: Prospectively collected data on all patients who underwent RIMA harvesting between January 1995 and February 2002 were analyzed. Thirty-one patients with right phrenic nerve injury were identified and the medical charts reviewed. Phrenic nerve injury was diagnosed when a postoperative chest roentgenogram showed the right hemidiaphragm to be two or more intercostal spaces higher than the left, or transection of the nerve was seen intraoperatively. Investigations included fluoroscopy and spirometry in upright and supine positions. Diaphragm plication was offered for symptom control. Subsequent follow-up was undertaken to determine the incidence of spontaneous recovery of diaphragm function and the benefits of diaphragm plication. RESULTS: Seven hundred and eighty-three patients underwent high mobilization of the RIMA with proximal detachment for use as a free graft. Thirty one patients with right hemidiaphragm dysfunction were identified in the postoperative period providing an injury incidence of 4% (confidence interval, 2.6% to 5.3%). Of these, 12 patients underwent diaphragm plication (4 early and 8 late), 14 patients achieved spontaneous recovery, and 5 patients were lost to follow-up. The supine to upright forced vital capacity ratios at the time of phrenic nerve dysfunction, after diaphragm plication, and after spontaneous recovery were 0.79, 0.90, and 0.96 respectively. CONCLUSIONS: The incidence of phrenic nerve injury associated with high RIMA harvesting was 4% but spontaneous recovery may be anticipated in two thirds (14 of 22) of patients in whom the injury is identified postoperatively. High RIMA harvesting should be used with caution in patients with preoperative pulmonary dysfunction in whom phrenic nerve injury would be poorly tolerated. PMID- 12902083 TI - Pressure distension stimulates the expression of endothelial adhesion molecules in the human saphenous vein graft. AB - BACKGROUND: Mechanical trauma occurring during saphenous vein graft harvesting plays a major role in graft failure after coronary bypass surgery. There is increasing evidence that neutrophil-endothelial interaction is involved in the pathogenesis of early graft occlusion. This study evaluates the effect of pressure distension on the expression of endothelial adhesion molecules in human saphenous vein. METHODS: Segments of saphenous vein graft (SVG) were collected from 20 patients undergoing coronary bypass surgery. We evaluated the expression of intercellular adhesion molecule (ICAM-1), vascular cell adhesion molecule (VCAM-1), and P-selectin on SVG endothelium under basal conditions and after pressure distension at 300 mm Hg. In the same experimental setting we also evaluated adhesion of both unstimulated and activated neutrophils to the endothelium of SVG. RESULTS: Control endothelial cells exhibited only a weak staining for intercellular adhesion molecule (ICAM-1), vascular cell adhesion molecule (VCAM-1), and P-selectin, whereas the levels of adhesion molecules increased significantly in the distended veins. Similarly, significantly greater adhesion of both unstimulated and activated neutrophils was observed in distended veins compared with control veins. CONCLUSIONS: Pressure distension of SVG before coronary bypass surgery induces upregulation of endothelial adhesion molecules, with subsequent increase in neutrophil adhesion to the endothelium. Neutrophil adhesion to endothelial cells may contribute to early failure of SVG. PMID- 12902086 TI - The effect of left internal mammary artery utilization on short-term outcomes after coronary revascularization. AB - BACKGROUND: The purpose of this study was to determine whether use of the left internal mammary artery (LIMA) during coronary revascularization influences short term morbidity in all patients undergoing revascularization, as well as in patients over the age of 75 years, female patients, and patients with diabetes. The study also explored variability in the utilization of LIMA grafts across an entire state. METHODS: Using the Clinical Outcomes Assessment Program (COAP) of the state of Washington, procedural outcomes were compared for patients receiving and patients not receiving LIMA grafts as part of revascularization procedures from January 1, 1999 to December 31, 2000. Mortality and major complications were examined, both as unadjusted rates and after adjusting for baseline patient risk factors. RESULTS: A total of 16 centers performed 8,797 nonemergent coronary artery revascularizations, including 81.7% with LIMA grafts. The use of a LIMA graft was associated with a significantly lower mortality (3.7% No LIMA vs 1.6% LIMA), as well as decreases in ventricular arrhythmias, need for postoperative dialysis, need for transfusions, ventilator dependence, and length of hospital stay. These trends were true for the population as a whole as well as for all subgroups analyzed, and they persisted after correcting for differences in comorbid conditions. In addition, there was wide variability in the use of LIMA grafts from center to center in the state. CONCLUSIONS: The use of LIMA grafts for coronary revascularization is associated with decreased mortality and morbidity. Despite these advantages, there is great variability in its application across the state of Washington. PMID- 12902087 TI - The Ross procedure in adults: intermediate-term results. AB - BACKGROUND: The durability of the Ross procedure may be optimized by appropriate geometric matching of the aortic and pulmonary artery roots. We employed a surgical strategy to standardize the operation in order to avoid more readily a geometric mismatch. METHODS: The Ross procedure was performed as a root replacement. Without regard for patient body surface area, the aortic annulus was plicated to 23 mm and externally buttressed with felt. Geometric mismatch of the distal autograft anastomosis was avoided by liberal use of a synthetic interposition graft, and the anastomosis was also externally buttressed with felt. An over-sized pulmonary homograft (27 to 28 mm) was routinely used to reconstruct the right ventricular outflow tract. RESULTS: Forty-four consecutive patients (27 men and 17 women; mean age, 49 +/- 9 years) were operated on between January 1997 and March 2002. Mean follow-up was 38 +/- 5 months. Twenty-nine patients had aortic stenosis and 15 had aortic regurgitation. Aortic annular plication was done in 41 (93%) and an aortic interposition was used in 14 (32%). There were three hospital deaths, with no subsequent deaths. Only 1 patient required reoperation 2.5 years postoperatively from recurrent endocarditis. No patient has more that "trivial" autograft insufficiency, and the mean autograft gradient was 7 +/- 3 mm Hg. No patient has significant pulmonary homograft stenosis. CONCLUSIONS: Geometric matching of the aortic and pulmonary roots may be readily accomplished using a standardized approach to the Ross procedure. In turn, this may optimize the durability of the operation. PMID- 12902088 TI - Cardiac valve replacement in human immunodeficiency virus-infected patients. AB - BACKGROUND: Valve replacement in human immunodeficiency virus (HIV)-infected patients is being performed with increasing frequency, but the early and late results in these immunocompromised patients are not known. METHODS: A 10-year retrospective clinical review was undertaken; patients and their physicians were contacted for follow-up clinical status. RESULTS: Twenty-two HIV-infected patients underwent valve replacement between 1990 and 1999, with no operative or hospital deaths. Mean patient age was 37.6 years; 15 were men. Indications for operation were heart failure in 59% (13/22) and sepsis in 91% (20/22). There were 12 aortic valve replacements, seven mitral valve replacements, and three double valve replacements. Mechanical valves were used in 11, bioprostheses in seven, and homografts in four. Follow-up information was available in 20 of 22 patients (84%). At mean follow-up of 5 years, there were 10 late deaths, due to: intracerebral hemorrhage (2), heart failure (2), unknown cause (2), renal failure (1), AIDS (1), sepsis (1) and endocarditis (1). Of the 20 patients with active preoperative endocarditis, 4 (20%) developed recurrent endocarditis; freedom from recurrent endocarditis was 83% at 1 year. Intravenous drug abuse was reported in 16 patients; survival among these patients was 94% at 1 month and 50% at 5 years. Recurrent endocarditis was only seen in patients with continued intravenous drug abuse. CONCLUSIONS: Valve replacement in HIV-infected patients has low operative risk, but late results are poor when HIV infection is associated with intravenous drug abuse, probably due to immunocompromise and continued high-risk behavior. PMID- 12902089 TI - Is 30-day mortality an adequate outcome statistic for patients considering heart valve replacement? AB - BACKGROUND: In-hospital mortality is widely used by clinicians as a benchmark measure of outcome for determining risks/benefits of cardiac surgery. Patients, however, may wish to have information on estimated longer-term outcomes. Mortality risk by 1 year after the operation may be a more meaningful outcome statistic. We therefore undertook to determine 30-day and 365-day postoperative mortality rates in a large series of consecutive patients who have undergone heart valve replacement (HVR) surgery in the United Kingdom since 1986. METHODS: Data on 80,757 patients registered on the UK Heart Valve Registry were analyzed. Kaplan-Meier actuarial survival analysis was calculated to determine 30-day (group 1) and 365-day (group 2) mortality. Cox proportional hazards were calculated for each group to identify significant risk factors for mortality less than 1 year. RESULTS: Thirty-day mortality represents around half (56%) of the 365-day mortality. This ratio was robust for most subdivisions of the total population. Cox proportional hazards demonstrated female sex, age older than 70 years, single tricuspid valve replacement, multiple valve replacement regardless of type of valve, and subsequent valve operation as significant risk factors of early mortality (group 1). However, men were at significantly greater risk of late mortality (group 2). All other factors remained significant with the exception of subsequent valve operation, which was dropped from the model (group 2). CONCLUSIONS: The robustness of these data would allow cardiologists and cardiac surgeons to provide preoperative patients with a reasonably accurate estimate of survival rates at 1 year after valve replacement surgery in addition to the customary short-term 30-day outcome. PMID- 12902090 TI - Invited commentary. PMID- 12902091 TI - Bivalvular mechanical mitral-aortic valve replacement in 254 patients: long-term results--a 22-year follow-up. AB - BACKGROUND: We have retrospectively studied 254 patients who underwent a bivalvular mechanical mitral-aortic replacement in the cardiovascular and thoracic surgery unit of Nantes from 1979 to 1989. The follow-up was 22 years (1979 to 2001). The last patient was operated on 12 years before the end of the follow-up. METHODS: All mitral prostheses were St. Jude Medical (SJM) bileaflet valves, and the aortic prostheses were 124 monodisc Bjork-Shiley valves, 3 Sorin prostheses, and 127 St. Jude Medical bileaflet prostheses. The mean age was 56.8 +/- 8.5 years with a sex ratio equal to 1. Rheumatism as the etiology predominated with 79.5%. Ninety-seven percent of the patients were followed for a total of 2,779 patient-years and a mean of 11.7 years. RESULTS: Operative mortality was 7.08%. Freedom from overall mortality and valve-related mortality at 22 years were 45.7% +/- 3.6% and 73.1% +/- 3%, respectively. The linearized rates of thromboembolic and hemorrhagic events were 1.07% and 0.9% per patient year, respectively. Multivariate analysis showed age (p < 0.002), sex (p < 0.01), and degenerative etiology (p = 0.04) as independent factors of late mortality, and age, sex, degenerative disease, and tricuspid pathology were related to valve related mortality. CONCLUSIONS: This study shows good results after mechanical mitral-aortic replacement in terms of survival rate and quality of life in surviving patients, and outlines the factors influencing long-term results as compared with isolated mitral valve replacement. PMID- 12902092 TI - Emergent endovascular stent grafting for perforated acute type B dissections and ruptured thoracic aortic aneurysms. AB - BACKGROUND: The purpose of our study was to demonstrate the effectiveness of endovascular stent grafts in the treatment of acutely ruptured thoracic aortic aneurysms and type B dissections as an alternative to the conventional surgical approach in an emergency setting. METHODS: From January 2001 to October 2001, we deployed 11 emergent endovascular stent grafts into the thoracic aorta. We treated seven ruptured aortic aneurysms and four acutely perforated type B dissections. Aortic rupture was confirmed preoperatively by spiral computed tomography. In all cases, hemothorax was present. The average interval from onset of symptoms to treatment was 28.5 hours. We used nine Talent and two Excluder stent grafts. RESULTS: Deployment of the stent grafts was successful in nine cases. There were two cases of access failure due to small caliber of iliac arteries, and 1 of these patients died shortly after the procedure was abandoned, At 12 months of follow-up, there were no cases of paraplegia, stent migration, or endoleaks. There was, however, one temporary renal failure, and 2 patients required mechanical ventilation for more than 48 hours. CONCLUSIONS: Our experiences with emergency endovascular stent grafting show that the procedure is technically feasible, with less morbidity and mortality than conventional open surgery, in high-risk patients. PMID- 12902093 TI - A double patch sandwich technique for surgical repair of acute aortic dissection type A. AB - BACKGROUND: This study compares two groups of patients with acute aortic dissection type A in whom two different techniques of anastomotic reinforcement were used, and evaluates the impact of these two techniques on perioperative blood loss and surgical outcome. METHODS: One hundred eighty-five consecutive patients with acute aortic dissection type A between 1998 and 2002 were grouped according to the technique utilized for reinforcing the aortic anastomotic site. Group A consisted of 21 patients in whom a novel double patch sandwich technique was used, whereas in group B (164 patients) conventional Teflon felt strips served as reinforcement. RESULTS: Preoperative profiles were comparable for both groups. Deep hypothermia and circulatory arrest were used in 180 patients (97%). Retrograde cerebral perfusion and retrograde cardioplegia were used in 70 patients (38%). Hospital mortality for the two groups was 4.7% (1 of 21) and 18% (29 out of 164), p < 0.30, respectively. On average 2 versus 6 U of PRBC (p < 0.21), and 3 versus 5 U of FFP (p < 0.004) were given during operation in groups A and B, respectively, and indicates reduced suture line bleeding in group A. CONCLUSIONS: Modifications in the surgical technique for repair of acute aortic dissection type A, and in anastomotic reinforcement in particular, may lead to substantial reduction of suture line bleeding and diminished blood loss and transfusion requirements, and favorably affects patient outcome. Nevertheless, continued effort is mandatory to further enhance surgical outcome in this patient population. PMID- 12902094 TI - Readmission to the intensive care unit after "fast-track" cardiac surgery: risk factors and outcomes. AB - BACKGROUND: The introduction of "fast-track" management into cardiac surgery has significantly shortened the intensive care unit (ICU) length of stay. Readmission to the ICU, traditionally used as a quality index, has not been investigated in these patients. The aim of this study was to assess the causes, risk factors, and outcomes associated with readmission to the ICU. METHODS: All patients undergoing open-heart surgery in a tertiary care, university-affiliated center were included in this prospective observational study. Preoperative and intraoperative data as well as ICU outcome were noted in all patients. RESULTS: Over the 27-month study period,1,613 patients were targeted for fast track management (discharge from ICU on the first postoperative day). The readmission rate was 3.29% (53 patients). Forty-three percent of readmissions occurred within 24 hours of discharge usually because of pulmonary problems (43%) or arrhythmias (13%). Readmission was associated with a prolonged ICU stay (105 +/- 180.0 versus 19.2 +/- 2.4 hours of initial ICU stay) and worse outcome: the only patients who died (6 of 53, 11.3%) were in this group. On multivariate analysis, a Bernstein-Parsonnet risk estimate more than 20 strongly predicted readmission (odds ratio, 3.08; 95% confidence interval, 1.43 to 6.69). CONCLUSIONS: Among a homogeneous group of patients targeted for fast-track management after cardiac surgery, readmission although uncommon is associated with a longer second ICU stay and significant mortality. The recognition of specific risk factors may allow for appropriate modification of the postoperative course. PMID- 12902095 TI - Emboli capture using the Embol-X intraaortic filter in cardiac surgery: a multicentered randomized trial of 1,289 patients. AB - BACKGROUND: Particulate emboli are thought to play a significant role in the development of cardiac surgical complications. Intraaortic filtration of particulate emboli may reduce the burden of this morbidity in cardiac patients. METHODS: A multiinstitutional randomized trial was designed and enrolled 1,289 patients at 22 centers. Six hundred forty-five patients were assigned to the treatment arm and received the Embol-X intraaortic filter, whereas 644 patients were assigned to the control arm. The endpoints examined were mortality, stroke, transient ischemic attack, renal insufficiency/failure, myocardial infarction, gastrointestinal complications, and limb-threatening ischemia. All filters were examined for histologic evidence of particulate emboli. RESULTS: Particulate emboli were identified in 598 (96.8%) of 618 filters successfully deployed. Composite event rates for the clinical endpoints were similar in both the filtered and nonfiltered arm (110/645 = 17% vs 122/644 = 19%, respectively). Individual event rates were also similar in both arms. A post hoc comparison of patients with moderate or greater preoperative risk scores demonstrated event reduction favoring the filtered group for renal complications (17/124 = 14% vs 28/117 = 24%, p = 0.04) and for the composite endpoint (30/124 = 24% vs 42/117 = 36%, p = 0.047). No clinically evident complications attributed to the use of the filter were identified. CONCLUSIONS: The use of the Embol-X intraaortic filter is both safe and effective, as demonstrated by the emboli capture rate of 97%. In addition, post hoc analysis indicates a reduction in postoperative renal complications for patients with moderate or greater preoperative risk. Further study of high-risk patients is warranted. PMID- 12902096 TI - Perfusing and ventilating the patient's lungs during bypass ameliorates the increase in extravascular thermal volume after coronary bypass grafting. AB - BACKGROUND: To test the hypothesis that bilateral extracorporeal circulation (ECC) (Drew technique) ameliorates the increase in extravascular thermal volume (ETV) observed after conventional cardiopulmonary bypass (CPB) in patients undergoing coronary artery bypass grafting. METHODS: Thirty-four consecutive patients underwent either bilateral ECC (n = 24, additional cannulation of pulmonary artery and left atrium and lungs perfused and ventilated during bypass) or conventional CPB (n = 10, right atrial and aortic cannulation, lungs statically inflated to 4 mbar (0.41 cm H(2)O) with oxygen, 500 mL/min). Determinations of ETV (thermodye dilution technique) and intraoperative fluid balance were made before surgery, at the end of surgery, and 4 hours thereafter. In addition, interleukin (IL)-8, thromboxane B2 (TxB(2)), and endothelin (ET)-1 concentrations were measured in the right atrium and pulmonary vein at specified time points. RESULTS: Comparisons of ETV made at the start of surgery, after aortic declamping, and after termination of ECC, respectively, revealed an increase from 4.8 +/- 0.2 mL/kg (mean +/- SEM) to 6.7 +/- 0.4 mL/kg, and 6.3 +/- 0.3 mL/kg with conventional CPB but ETV remained unchanged at 5.2 +/- 0.3 mL/kg, 5.1 +/- 0.2 mL/kg, and 4.9 +/- 0.3 mL/kg with bilateral ECC. Priming volume (1,580 +/- 10 mL versus 2,213 +/- 77 mL, p < 0.001) and intraoperative fluid balance (+1,955 +/- 233 mL versus +2,654 +/- 210 mL, p < 0.05) were less with conventional CPB. Concentrations of IL-8, TxB(2), and ET-1 were not different between groups. CONCLUSIONS: Despite a significantly greater prime volume and a more positive intraoperative fluid balance, ETV did not change with bilateral ECC but increased with conventional CPB. Thus, using the patient's lungs as an oxygenator during bypass mitigates the increase in extravascular pulmonary fluid. PMID- 12902098 TI - Impaired oxygenation and increased hemolysis after cardiopulmonary bypass in patients with glucose-6-phosphate dehydrogenase deficiency. AB - BACKGROUND: The purpose of this study was to determine whether the damaging effects of cardiopulmonary bypass, ischemia, and reperfusion would be more pronounced in patients with glucose-6-phosphate dehydrogenase deficiency undergoing cardiac surgery. METHODS: Forty-two patients with glucose-6-phosphate dehydrogenase deficiency underwent open heart procedures using cardiopulmonary bypass. This group was matched with a control group of identical size for comparison of operative course and postoperative outcome. The perioperative variables were compared between the two groups using univariate and multivariate analysis. RESULTS: The duration of ventilation after the operation was significantly longer in the glucose-6-phosphate dehydrogenase-deficient group (13.7 +/- 7.6 hours versus 7.7 +/- 2.8 hours; p < 0.0001). Minimal value of arterial oxygen tension was lower in patients with glucose-6-phosphate dehydrogenase deficiency (66 +/- 12 mm Hg versus 85 +/- 14 mm Hg; p < 0.0001), and more cases of hypoxia (arterial oxygen tension < 60 mm Hg) were found in this group (11 versus 1; p = 0.001). Compared with the control group, patients with glucose-6-phosphate dehydrogenase deficiency had significantly elevated hemolytic indices expressed by bilirubin levels (26 +/- 10 mmol/L versus 17 +/- 6.7 mmol/L; p < 0.0001) and lactic dehydrogenase levels (970 +/- 496 U/L versus 505 +/- 195 U/L; p < 0.0001). They also required significantly more blood transfusion perioperatively (1.9 +/- 1.4 packed cell units/patient versus 0.8 +/- 1.0 packed cell units/patient; p = 0.0001). CONCLUSIONS: Patients with glucose-6-phosphate dehydrogenase deficiency who are undergoing cardiac surgery may have a more complicated course with a longer ventilation time, more hypoxia, increased hemolysis, and a need for more blood transfusion. Because this difference may be caused by subnormal free radical deactivation, strategies that minimize bypass in general and free radicals specifically may be beneficial. PMID- 12902099 TI - Cardiomyocyte apoptosis and ischemic preconditioning in open heart operations. AB - BACKGROUND: The aim of the present study was to ascertain the percentage of left apical myocardial apoptosis in three-vessel coronary artery bypass grafting patients quantitatively and the impact of ischemic preconditioning. METHODS: Twenty-one patients with three-vessel disease who had elective coronary artery bypass grafting were randomized in a ratio of 2:1 to ischemic preconditioning (n = 14) or a control group (n = 7). The ischemic preconditioning protocol was established by two cycles of ascending aorta occlusion for 2 minutes followed by 3 minutes of reperfusion. Myocardial samples from the apex of the left ventricle were taken using a Tru-Cut needle before aortic cross-clamping and immediately after declamping. The percentage of apoptosis was analyzed by TUNEL methods. Data on hemodynamics and biochemical markers were collected. RESULTS: Low levels of myocardial apoptosis were found before the operation (0.01% +/- 0.00%). During the early reperfusion period, the percentage of myocardial apoptotic cells significantly increased (0.15% +/- 0.05%, p = 0.008). Ischemic preconditioning significantly improved cardiac index and right ventricular ejection fraction recovery after the operation (p = 0.036 and 0.001 respectively, repeated measure) but had no effect on myocardial apoptosis before and after the operation (0.01 +/ 0.00 versus 0.01 +/- 0.00, p = 0.658 and 0.12% +/- 0.04% versus 0.23% +/- 0.14%, p = 0.302). CONCLUSIONS: Cardioplegic myocardial ischemia during open heart operation was associated with induction of cardiomyocyte apoptosis in humans. Attenuation of postoperative cardiac dysfunction by ischemic preconditioning appeared to be independent of apoptosis. PMID- 12902100 TI - Acute circulatory actions of intravenous amiodarone loading in cardiac surgical patients. AB - BACKGROUND: The duration, severity, and cause of hypotension after intravenous amiodarone has not been well characterized in anesthetized cardiac surgical patients. Because amiodarone is tolerated in patients with advanced cardiac disease, we hypothesized that left ventricular systolic performance is preserved despite hypotension during amiodarone loading. METHODS: In a prospective double blind trial, 30 patients undergoing coronary artery bypass graft (CABG) surgery were randomly assigned to receive intravenous amiodarone (n = 15) or placebo (n = 15). Cardiac output (CO), mixed venous oxygen saturation (SVO), arterial blood pressure (systolic blood pressure [SBP], diastolic blood pressure [DBP], mean arterial pressure [MAP]), pulmonary artery pressure, and central venous pressure (CVP) were recorded. Transesophageal echocardiographic left ventricular end diastolic area (EDA), end-systolic area (ESA), fractional area change (FAC), and end-systolic wall stress (ESWS) were measured every 5 minutes. RESULTS: Mean arterial pressure, SBP, and DBP decreased over time after drug administration in both groups (p < 0.05). At 6 minutes, amiodarone decreased the MAP by 14 mm Hg (p = 0.004) and placebo decreased the MAP by 4 mm Hg. The change in MAP, SBP, and DBP between groups was statistically different for the first 15 minutes after drug administration. Hypotension requiring intervention occurred in 3 of 15 after amiodarone and 0 of 15 after placebo (p = 0.22). The mean heart rate was 11.5 beats per minute less after amiodarone (p < 0.02), but pulmonary artery pressure, CVP, SVO, and FAC were not different between groups. CONCLUSIONS: Intravenous amiodarone decreased heart rate and caused a significant, but transient decrease in arterial pressure in the first 15 minutes after administration. Left ventricular performance was maintained suggesting that selective arterial vasodilation was the primary cause of drug-induced hypotension. PMID- 12902101 TI - Beyond Fontan conversion: Surgical therapy of arrhythmias including patients with associated complex congenital heart disease. AB - BACKGROUND: Arrhythmia operations may be extended to patients with failed ablation procedures or associated structural defects requiring surgical intervention. The purpose of this study is to review our experience with arrhythmia operations in 29 patients who did not have Fontan conversions after the introduction of catheter ablation. METHODS: Between July 1992 and January 2002, 29 patients had operations for refractory atrial (n = 24) or ventricular (n = 5) arrhythmias. Mechanisms of arrhythmia included atrial reentry (n = 11), atrial fibrillation (n = 5), automatic atrial (n = 3), accessory connections (n = 6), atrioventricular nodal reentry (n = 2), and ventricular tachycardia (n = 5). Median age at operation was 12.3 years (range, 6 days to 45 years). Two patients had structurally normal hearts; the remaining 27 patients underwent concomitant repair of structural heart disease, including atrioventricular valve replacement or repair (n = 8), anatomy-specific repair of Ebstein's anomaly (n = 4), tetralogy of Fallot repair or revision (n = 4), atrial septal defect closure (n = 3), ventricular septal defect repair (n = 2), Mustard takedown with arterial switch (n = 2), initial Fontan (n = 2), right ventricle-to-pulmonary artery conduit revision (n = 2), Norwood procedure (n = 1), 1 ventricular repair for Uhl's anomaly (n = 1), Mustard baffle revision (n = 1), pulmonary valve replacement with aneurysm resection (n = 1), and aortic valve replacement with complex repair (n = 1). RESULTS: No patient developed heart block, and the surgical mortality rate was 7%. One patient died after Mustard takedown and arterial switch operation, and 1 neonate died after repair of severe Ebstein's anomaly. There was one late death after arterial switch conversion at another institution. Recurrent clinical supraventricular tachycardia was present in 2 patients (2 of 27, 7.4%) and 2 patients had new-onset tachycardias with different underlying mechanisms of arrhythmia at late follow-up (median follow-up 47 months). CONCLUSIONS: Successful surgical therapy of arrhythmias can be performed safely at the time of repair of complex congenital heart disease or in patients with failed catheter ablation procedures. Early consideration for single-stage therapy of arrhythmia and structural heart disease is indicated. PMID- 12902102 TI - Does right ventricular outflow tract damage play a role in the genesis of late right ventricular dilatation after tetralogy of Fallot repair? AB - BACKGROUND: The aim of this study was to determine the relative role of pulmonary insufficiency and right ventricular outflow tract damage in the genesis of late symptoms related to right ventricular dilatation. METHODS: In a retrospective study we compared the late outcomes of patients who had undergone operations known to generate pulmonary insufficiency, namely, transventricular repair of tetralogy of Fallot and pulmonary commissurotomy for isolated pulmonary stenosis. RESULTS: In our institution, between 1964 and 1984, a total of 44 patients were found to have had an isolated pulmonary commissurotomy and 189 survived a transventricular repair of tetralogy of Fallot. Of these patients, 134 had patching of the right ventricle and 55 direct closure of a right ventriculotomy. Follow-up was 94% complete after a mean of 22 +/- 7 years. On echocardiography, patients with isolated commissurotomy had similar degrees of moderate and severe pulmonary insufficiency as tetralogy of Fallot patients who had a right ventricular patch (p > 0.2). However, freedom from adverse events related to right ventricular dilatation was far better (log rank p < 0.001) in patients with isolated commisurotomy. CONCLUSIONS: Pulmonary insufficiency is not the only determinant of late symptomatic right ventricular dilatation after repair of tetralogy of Fallot. Pulmonary insufficiency seems much more deleterious in patients who have had right ventricular outflow tract patching. Long-term pulmonary insufficiency alone is responsible for a slight degree of right ventricular dilatation, but symptoms may develop much later if the contractility of the pulmonary infundibulum is preserved. The pulmonary infundibulum may be essential for right ventricular ejection, and for maintaining pulmonary valve competence. PMID- 12902104 TI - Ventricular mechanics in the bidirectional glenn procedure and total cavopulmonary connection. AB - BACKGROUND: The time course of ventricular efficiency in Fontan candidates who underwent both the bidirectional Glenn procedure (BDG) and total cavopulmonary connection (TCPC) were analyzed in this study. We previously reported that volume load reduction of BDG preceding TCPC allowed for any afterload mismatch to be corrected, thereby improving ventricular efficiency after staged TCPC. METHODS: We measured percent normal systemic ventricular end-diastolic volume (%N-EDV), contractility (end-systolic elastance [Ees]), afterload (effective arterial elastance [Ea]), and ventricular efficiency (ventriculoarterial coupling [Ea/Ees]) based on cardiac catheterization data before and after both BDG and staged TCPC in 30 patients. Ees and Ea were approximated as follows: Ees = mean arterial pressure/minimal ventricular volume, and Ea = maximal ventricular pressure/(maximal ventricular volume - minimal ventricular volume), and Ea/Ees was then calculated. Ventricular volume was divided by body surface area. RESULTS: The %N-EDV decreased both after BDG and after staged TCPC, thus resulting in an improvement of Ees. Although Ea increased both after BDG and after staged TCPC, Ea decreased during the interval between BDG and staged TCPC. These changes resulted in an improvement in Ea/Ees during the interval period and after staged TCPC, although Ea/Ees worsened after BDG. CONCLUSIONS: Correction of afterload mismatch during the interval period between BDG and staged TCPC is considered to be one of the most important factors for obtaining excellent clinical results when selecting a staged strategy to treat high-risk Fontan candidates. PMID- 12902105 TI - Deletion of chromosome 22q11.2 and outcome in patients with pulmonary atresia and ventricular septal defect. AB - BACKGROUND: The 22q11.2 deletion (del22q) is present in many patients with conotruncal abnormalities including pulmonary atresia with ventricular septal defect (PA/VSD). We sought to determine the impact of the del22q on outcome in subjects with PA/VSD. METHODS: We reviewed the experience for all patients with PA/VSD who were born between January 1993 and April 2002 and presented to our institution. Patients with conotruncal defects were routinely evaluated for genetic disorders including del22q. Fluorescence in situ hybridization was used to test for del22q. RESULTS: There were 67 subjects with PA/VSD who presented during that time period; testing for del22q was performed in 58 of 67 (87%) and these 58 patients comprised the study population. The 22q11.2 deletion was present in 20 of 58 (34%) patients tested. Major aortopulmonary collaterals were defined by angiography and were present in 27 (47%). These collaterals were significantly more common among subjects with del22q (13 of 20, 65%; p = 0.04). The median cross sectional area of the pulmonary arteries, the Nakata index, was significantly less for patients with del22q (41 versus 142 mm(2)/m(2); p = 0.006). There were 3 subjects, all of whom had del22q, who did not undergo surgery owing to markedly hypoplastic pulmonary arteries. Of the remaining 55 patients, 53 had arteriopulmonary shunt with or without unifocalization as the initial procedure and 35 patients have undergone complete repair. There were 8 operative deaths and 1 nonoperative death. The 5-year survival was 36% for patients with del22q versus 90% for patients without del22q. The 22q11.2 deletion was a significant risk factor for death, even after adjusting for the presence of major aortopulmonary collaterals (p = 0.004). There was no significant difference between the two groups with respect to the incidence of serious viral, bacterial, or fungal infections in the perioperative period. CONCLUSIONS: Patients with del22q and PA/VSD are at increased risk for death owing to a variety of factors including less favorable pulmonary artery anatomy. A better understanding of del22q, pulmonary artery anatomy, and outcome is required. PMID- 12902106 TI - Implications of anomalous right subclavian artery in the repair of neonatal aortic coarctation. AB - Spinal cord perfusion is predominantly from the anterior spinal artery, which arises from the vertebral arteries by way of the subclavian arteries. Anomalous origin of the right subclavian artery and coarctation of the aorta is considered to be an increased risk factor for spinal cord damage, possibly because of the minimal collateral circulation during aortic clamping. The aim of this study is to review 5 consecutive cases of neonatal aortic coarctation with ARSA. METHODS: Five neonates (0.8 to 4.6 kg) underwent operation between July 1999 and December 2000 with resection of the coarctation and end-to-end anastomosis. Both subclavian arteries (n = 5) and left carotid artery (n = 4) were clamped, leaving the right carotid artery as the sole provider of perfusion for the spinal cord. RESULTS: Despite clamping of both subclavian arteries, right radial artery pressure was measurable in 4 of the 5 cases. Aortic cross-clamp times varied from 12 to 26 minutes at a core temperature of 34 degrees to 35 degrees C. There was no operative mortality. None of the neonates developed any major neurologic sequelae. CONCLUSIONS: When clamping the two subclavian arteries during coarctation repair, the spinal artery is left with collateral blood flow that can theoretically originate from the carotid arteries through the circle of Willis and retrogradely down the vertebral arteries. The presence of such collateral circulation was documented as recordable blood pressure in the right radial artery during surgical repair. PMID- 12902107 TI - Early postoperative prediction of cerebral damage after pediatric cardiac surgery. AB - BACKGROUND: Cerebral damage is a serious complication of pediatric cardiac surgery. Early prediction of actual risk can be useful in counseling of parents, and in early diagnosis and rehabilitation therapy. Also, if all children at risk could be identified therapeutic strategies to limit perioperative cerebral damage might be developed. The aim of this study is to create a mathematical model to predict risk of neurologic sequelae within 24 hours after surgery using simple and readily available clinical measurements. METHODS: The hospital records of 534 children after cardiac surgery were reviewed. Variables examined were age at operation, diagnosis, use of cardiopulmonary bypass, arterial and central venous oxygen saturation, serum glucose, lactate and creatine kinase, mean arterial pressure, and body temperature. The endpoint for each study patient was the occurrence or lack of occurrence of seizures, movement or developmental disorders, cerebral hemorrhage, infarction, hydrocephalus, or marked cerebral atrophy. Univariate and multivariate regression analyses were used to evaluate the predictive power of the investigated factors as well as to create a predictive model. RESULTS: In 6.26% of children symptoms of cerebral damage were found. Significant risk factors were age at surgery, more complex malformations, metabolic acidosis, and increased lactate (odds ratio: age, 0.882/yr [0.772 1.008]; complex malformations, 10.32 [1.32-80.28]; arterial pH more than 7.35 to 0.4 [0.18-0.89]; lactate -1.018 per mg/dL [1.006-1.03]). CONCLUSIONS: It is possible to quantify the risk of appearance of symptoms of cerebral damage after cardiac surgery within 24 hours using simple and readily available clinical measurements. PMID- 12902108 TI - Anomalous coronary artery origin from the pulmonary artery: correlation between surgical timing and left ventricular function recovery. AB - BACKGROUND: This study investigates the correlation between surgical timing and 15-year longitudinal left ventricular and mitral valve function, after repair of anomalous coronary artery origin from the pulmonary artery. METHODS: Between 1987 and 2002, 31 patients (median age, 7.1 months) underwent repair for anomalous origin of the left (n = 28), right (n = 2), or both (n = 1) coronary arteries from the pulmonary artery. Repair was accomplished by subclavian interposition in 5 patients, intrapulmonary tunnel in 12, and direct aortic reimplantation in 14. Primary mitral valve repair was never associated with coronary revascularization. Total follow-up was 186.4 patient-years (mean, 77.2 months). RESULTS: Fifteen year actuarial survival was 92.9% +/- 4.9% for coronary transfer, 40.0% +/- 21.9% for subclavian interposition, and 89.9% +/- 7.5% for intrapulmonary tunnel (p = 0.019). Five patients required further intervention for supravalvular pulmonary stenosis (n = 3), baffle leak (n = 1), and mitral valve replacement (n = 1). Coronary transfer allowed best freedom from long-term reoperation (92.3% +/- 7.4%). Left ventricular shortening fraction increased from 17.3% +/- 6.3% before operation to 34.1% +/- 4.6% at last follow-up (p < 0.01). Regression analysis demonstrated a linear relationship between age at repair and shortening fraction recovery (r(2) = 0.573, p < 0.01). Patients younger than 6 months of age showed worse preoperative shortening fraction (15.9% +/- 5.2%) and best longitudinal shortening fraction recovery (36.4% +/- 5.1%; p < 0.001). Major improvement in mitral valve function was observed within 1 year from surgery in 90.4% of survivors. CONCLUSIONS: Repair of anomalous coronary artery origin from the pulmonary artery in younger symptomatic infants offers the best potential for recovery of left ventricular function, despite a worse initial presentation. Coronary transfer is associated with superior long-term survival and freedom from reoperation. Most patients with patent two-coronary repair will recover normal mitral valve function; therefore, simultaneous mitral valve surgery seems unwarranted. PMID- 12902111 TI - Saber-sheath malacic trachea remodeled and fixed into a normal shape by long-term placement and then removal of gianturco wire stent. AB - We report the case of a 59-year-old man who presented with major dyspnea due to saber-sheath malacic trachea associated with chronic pulmonary obstructive disease. The placement of a temporary tracheal stent alleviated his dyspnea very well; hence the stent was replaced with a Gianturco wire stent (Cook Cardiology, Bloomington, IN). However, this required removal due to wire-stent-related complications 2 years after the replacement. Surprisingly the trachea had been remodeled to a normal shape resulting in comfortable, functional respiratory status. A review of the literature reveals our case to be the first report of curing saber-sheath malacic trachea without leaving any prostheses or other foreign materials. PMID- 12902110 TI - Outcome of unroofing procedure for repair of anomalous aortic origin of left or right coronary artery. AB - BACKGROUND: Anomalous aortic origin of a coronary artery (AAOCA) from an incorrect sinus of Valsalva is a relatively rare congenital defect and is associated with sudden death. Several surgical techniques have been described to address this defect, but functional outcome has never been addressed. In this report we evaluate a consecutive series of patients treated with unroofing techniques using transthoracic echocardiography, graded exercise testing, and stress echocardiography to assess functional repair. METHODS: Nine consecutive patients (range 7 to 65 years) underwent surgical repair of AAOCA from 1995 to 2001. In all patients the intramural segment was at or below the level of the commissure. All patients underwent a modified unroofing procedure to move the coronary artery orifice to the appropriate sinus. In 2 patients, a new orifice was created without significant unroofing and disruption of the commissure. Patients were evaluated prospectively with exercise electrocardiography testing and by resting and stress echocardiography. RESULTS: Of the 9 patients, 8 presented with symptoms suggestive of ischemia (chest pain, dyspnea on exertion, or syncope). Six patients had anomalous left main coronary artery arising from the right sinus of Valsalva, and 3 patients had anomalous right coronary artery from the left sinus of Valsalva. Transthoracic echocardiography and graded exercise testing was performed in all 9 patients (mean 29 months, range 4 to 85 months) after repair. Of the 9 patients, 8 also underwent stress echocardiography. In 8 of 9 patients the newly created coronary artery ostium was visualized by either two-dimensional echocardiography or color flow Doppler. All patients were symptom free at the time of follow-up. Exercise stress echocardiography was negative in all patients. Of the 8 patients, 7 had normal left ventricular shortening. No patients had regional wall motion abnormalities suggestive of ischemia. All patients were intervention free except 1 patient who developed severe aortic insufficiency and underwent a subsequent Ross procedure 44 months after his initial procedure. CONCLUSIONS: Anomalous origin of a coronary artery from an incorrect sinus of Valsalva is known to be associated with increased risk of sudden death. Surgical correction can be carried out with minimal risk and good anatomic and functional results. Manipulation of the commissure can be avoided by creation of a neo-ostia without extensive unroofing of the intramural segment or manipulation of the intercoronary commissure. This may avoid aortic valve malfunction. PMID- 12902112 TI - Pulmonary embolism after lung resection: diagnosis and treatment. AB - Pulmonary embolism after lung resection (PEALR) has a high mortality rate, and it is one of the most severe complications after lung resection. Early diagnosis and treatment are essential for PEALR. Here we present 3 cases of severe PEALR. In these cases, transthoracic Doppler echocardiography was useful for confirming the diagnosis of PEALR. Thrombolysis with recombinant tissue plasminogen activator (r tPA) was used to treat the embolism, and these patients were subsequently discharged. Thus echocardiography may become a primary procedure to confirm the diagnosis of severe PEALR, and thrombolysis with second-generation r-tPA may be the preferred choice for treatment. PMID- 12902114 TI - Primary lymphoepithelioma of the esophagus. AB - A case of primary lymphoepithelioma of the esophagus is presented in an Arabic female patient. This very rare Epstein-Barr virus infection-related malignancy has previously been reported only in patients from Japan. The tumor exhibited classic histologic and immunohistochemical features of lymphoepithelioma, and was successfully treated with neoadjuvant chemotherapy and irradiation followed by surgical resection. PMID- 12902113 TI - Pulmonary inflammatory myofibroblastic tumor invading the left atrium. AB - Inflammatory myofibroblastic tumor is a rare solid tumor that most often affects children and young adults. Although benign, the tumor may be very aggressive locally. We describe a 9-year-old boy with primary inflammatory myofibroblastic tumor of the left upper lobe involving the left atrium. PMID- 12902115 TI - A case of fatal West Nile virus meningoencephalitis associated with receipt of blood transfusions after open heart surgery. AB - First identified in the United States in 1999, West Nile virus caused approximately 3,500 infections in the late summer and fall of 2002. The virus is predominantly transmitted by mosquitoes, and the risk of infection through blood product transfusion is believed to be low. We present a case of West Nile virus encephalitis transmitted by red blood cell transfusion at the time of coronary artery bypass grafting that resulted in the patient's death. Individuals undergoing procedures with high blood product transfusion requirements, such as cardiac surgery or organ transplantation, may be at higher risk of this nosocomial infection during epidemics. PMID- 12902116 TI - Methotrexate in acute persistent humoral rejection: an option for graft rescue. AB - Humoral response emerges as an important component in acute graft rejection and a new challenge to clinicians in posttransplant care. Management of recurrent episodes and persistent activation of the humoral component of the immune system, despite the usual therapeutic approach to rejection, remains unknown. This article describes the successful use of methotrexate as an option for rescuing a graft in this worrisome situation. PMID- 12902117 TI - Successful management of empyema in a patient with a total artificial heart. AB - A description of successful management of a patient who developed an empyema as a postoperative complication following the insertion of a CardioWest total artificial heart (TAH) as a bridge to cardiac transplantation is presented. By using traditional methods of management, the patient recovered and went on to transplant. PMID- 12902118 TI - A strategy of hypothermic circulatory arrest for difficult heart transplant postventricular assist device. AB - Donor heart and ventricular assist device excision can be extremely difficult at the time of heart transplant. We present a strategy of hypothermic circulatory arrest established with ventricular assist device cannulas for difficult heart transplants. The device inlet or outlet cannulas already in place, or both, are used to complement the safe cannulation sites available. This approach permits controlled excision of the recipient heart and device, easy access to convert to standard ascending aorta and bicaval cannulation, and minimizes the donor graft anoxia time. Two case reports are presented. PMID- 12902119 TI - Subdural hemorrhagic injury after open heart surgery. AB - We report two cases of acute subdural hematoma after cardiac surgery using cardiopulmonary bypass. In both patients, emergency removal and drainage of a subdural hematoma was performed by neurosurgeons, and complete recovery followed. Subdural hemorrhagic brain injury after cardiac surgery is rare and devastating; however, we consider early diagnosis and proper treatment to be effective because organic brain damage did not occur. PMID- 12902120 TI - Off-pump right atrial thrombectomy for heparin-induced thrombocytopenia with thrombosis. AB - This report describes a 72-year-old woman with atrial fibrillation who presented with lower extremity ischemia secondary to thromboembolism. After lower extremity thrombectomy, the patient developed heparin-induced thrombocytopenia with thrombosis (HITT). Her postoperative course was complicated by recurrent supraventricular and ventricular tachycardia, secondary to a mobile thrombus in the right atrium extending into the right ventricle. Because administration of heparin was contraindicated, the patient underwent off-pump right atrial thrombectomy during a brief period of inflow occlusion. Postoperatively, she was placed on lepirudin. Her platelet count normalized without any further thrombotic episodes, and she was discharged on warfarin. PMID- 12902122 TI - Uncommon presentation and surgical correction of unroofed coronary sinus syndrome. AB - A 59-year-old man with signs and symptoms of congestive heart failure, occurring a few months after an infective episode, underwent cardiac investigations revealing severe biventricular dysfunction, persistent left superior vena cava with almost completely unroofed coronary sinus, and critical stenosis of the proximal right coronary artery. Surgical correction of the congenital malformation associated with revascularization of the right coronary allowed a prompt recovery of clinical conditions and ventricular function. PMID- 12902121 TI - Regression of intracardiac heparin-induced thrombosis after aortic root surgery. AB - Heparin-induced thrombocytopenia and thrombosis syndrome (type II) is associated with thromboembolic complications and a mortality rate up to 30%. We describe a patient who developed intracardiac and aortic Dacron prosthesis heparin-induced thrombosis after aortic root conservative surgery. Successive transoesophageal echocardiographies demonstrated a progressive regression of intracardiac thrombosis with oral anticoagulation by warfarin and antiplatelet therapy combining aspirin and clopidogrel. PMID- 12902123 TI - Concomitant transabdominal MIDCAB and abdominal aortic aneurysm repair. AB - We present 2 patients who underwent transabdominal minimally invasive direct coronary artery bypass with the right gastroepiploic artery combined with abdominal aortic aneurysm repair. The surgical procedures, both performed through a median laparotomy, proved safe and of limited invasiveness. The one-stage surgical intervention prevented catastrophic complications, such as acute myocardial infarction or rupture of abdominal aortic aneurysm. We believe that concomitant transabdominal minimally invasive direct coronary artery bypass and abdominal aortic aneurysm repair should be considered as a single combined surgical strategy in selected patients. PMID- 12902124 TI - Transatrial approach of acquired posterior ventricular septal rupture and double orifice technique in tricuspid valve repair. AB - We present a successful transatrial repair of ventricular septal rupture and tricuspid valve reconstruction, using the "edge-to-edge" technique, as a serious complication of a posterior myocardial infarction in an 83-year-old woman. PMID- 12902125 TI - Left ventricular thrombus sans overt cardiac pathology. AB - A 25-year-old man presenting with peripheral and cerebral emboli was incidentally detected as having a left ventricular mass on two-dimensional echocardiographic examination of the heart. In absence of any obvious structural heart disease or hypercoagulable state, the mass was presumptively diagnosed as left ventricular myxoma. The patient was operated on, and histopathology revealed the mass to be a thrombus. The cause and pathogenesis of the left ventricular thrombus, differential diagnosis, and management are discussed. The rarity of the case is highlighted. PMID- 12902126 TI - Intrapericardial teratoma in a neonate. PMID- 12902127 TI - Three-dimensional computed tomography of thoracic aortic aneurysm. PMID- 12902129 TI - Safe hemostasis by application of a new strict graft inclusion technique for replacement of the aortic root. AB - Bleeding from a coronary anastomosis and aortic annulus is sometimes a problem in the technique for aortic root replacement described by Bentall and DeBono and Cabrol and colleagues. Exposure and repair of these suture lines may be quite difficult after the termination of bypass. This article describes a new technique for aortic root replacement. In our experience to date, 6 patients with aneurysm of the ascending aorta and root and a calcified aortic valve stenosis as well as 1 patient with aortic dissection type A have benefited from this new procedure. PMID- 12902128 TI - Description of a new technique for reimplanting the anomalous right pulmonary artery. AB - Anomalous origin of the right pulmonary artery from the ascending aorta is a rare congenital lesion with a high mortality and morbidity if early diagnosis is not made and correction is not undertaken. We describe the repair of such a lesion using a double-trapdoor technique of pulmonary artery reimplantation. PMID- 12902130 TI - Technique of using the St. Jude aortic connector with the radial artery. AB - We present a technique for the use of St. Jude Aortic Connector System with the radial artery. This technique will allow surgeons, who prefer the radial artery to the vein, to use this approach whenever feasible. PMID- 12902131 TI - Off-pump long onlay bypass grafting using left internal mammary artery for diffusely diseased coronary artery. AB - Surgical treatment of a diffusely diseased coronary artery has been considered a relative contraindication for off-pump coronary artery bypass grafting. To our knowledge, long onlay-patch grafting with off-pump coronary artery bypass grafting has not been described. Two sets of Octopus 3 tissue stabilizers were placed longitudinally along the target coronary artery. This allowed us to perform surgical angioplasty and bypass grafting without cardiopulmonary bypass support (double Octopus technique). We report our early experience with off-pump long onlay bypass grafting in patients with a diffusely diseased coronary artery using double Octopus tissue stabilizers. Diffusely diseased coronary artery; off pump coronary artery bypass grafting; coronary artery reconstruction; coronary artery bypass grafting; onlay patch anastomosis PMID- 12902132 TI - Heparin-induced thrombocytopenia and cardiac surgery. AB - Unfractionated heparin given during cardiopulmonary bypass is remarkably immunogenic, as 25% to 50% of postcardiac surgery patients develop heparin dependent antibodies during the next 5 to 10 days. Sometimes, these antibodies strongly activate platelets and coagulation, thereby causing the prothrombotic disorder, heparin-induced thrombocytopenia. The risk of heparin-induced thrombocytopenia is 1% to 3% if unfractionated heparin is continued throughout the postoperative period. When cardiac surgery is urgently needed for a patient with acute or subacute heparin-induced thrombocytopenia, options include an alternative anticoagulant (bivalirudin, lepirudin, or danaparoid) or combining unfractionated heparin with a platelet antagonist (epoprostenol or tirofiban). As heparin-induced thrombocytopenia antibodies are transient, unfractionated heparin alone is appropriate in a patient with previous heparin-induced thrombocytopenia whose antibodies have disappeared. PMID- 12902133 TI - Epithelioid hemangioendothelioma and other vascular mediastinal tumors: a role for alpha-2a interferon? PMID- 12902136 TI - Time course of proinflammatory and anti-inflammatory responses after cardiac operation: monocyte HLA-DR expression. PMID- 12902137 TI - Counter-intuitive results from nonrandomized trials may unmask the "art of medicine". PMID- 12902139 TI - Is the hemodynamic performance of the carpentier-edwards perimount valve really equivalent to that of stentless valves? PMID- 12902141 TI - Pannus formation after mitral valve replacement with the omniscience prosthesis: letter 1. PMID- 12902142 TI - Omniscience valve results: letter 2. PMID- 12902143 TI - Coronary flow reserve in composite arterial Y grafts. PMID- 12902145 TI - Non-cardioplegic coronary bypass is effective, teachable, and still widely used: letter 1. PMID- 12902146 TI - Coronary surgery with intermittent aortic cross-clamping: a word of caution on the incidence of cerebrovascular accidents: letter 2. PMID- 12902148 TI - Expression of functionally relevant cell surface markers in dibutyltin-exposed human natural killer cells. AB - Butyltin (BT) compounds are known for their worldwide contamination. Dibutyltin (DBT) is used as a stabilizer in plastic products, and as a deworming agent in poultry. Poultry products have been shown to contain measurable levels of DBT. Drinking water has also been reported to contain BTs due to leaching from PVC pipes. We, and others, have found measurable levels of DBT in human blood. BTs appear to increase the risk of cancer and other viral infections in exposed individuals. In previous studies we have shown that the tumor killing function of natural killer (NK) lymphocytes was greatly diminished after as little as a 1 h exposure to DBT and the inhibition continued even after removal of the compound. We also showed that there was a significant decrease in NK cell lysis of K562 target cells after an exposure to 1.5 microM DBT for 24 h. This 24 h exposure also decreased the ability of NK cells to bind to tumor cells. Loss of binding function was not seen when NK cells were exposed to 5-10 microM DBT for 1 h. However, NK cells exposed to 5 microM DBT for 1 h and then incubated in DBT-free media for 24, 48, or 96 h, showed a significant loss of tumor-binding function within 24 h. The effects of DBT exposure on seven cell surface molecules that are involved in NK-cell interactions with target cells were investigated. The results indicated that the exposure of NK cells to 1.5 microM DBT for 24 h decreased the expression of CD2, CD11a, CD16, CD11c. There was no decrease in expression of any of the markers studied when NK cells were exposed to 5 microM DBT for 1 h, consistent with the fact that a 1-h exposure had no effect on the ability of NK cells to bind tumor cells. However, when NK cells were exposed to 5 microM DBT for 1 h followed by 24, 48 or 96 h incubations in DBT-free media there was decreased expression of several of the cells surface molecules with the most dramatic decreases being in CD16 and CD56. PMID- 12902149 TI - Structural activity relationship between Salmonella-mutagenicity and nitro orientation of nitroazaphenanthrenes. AB - Nitroazaphenanthrenes (NAphs) and their N-oxides (NAphOs) were synthesized as derivatives with nitrogen atoms in the 1, 4, and 9 positions of phenanthrene rings, and as nitrated derivatives substituted at the 1, 2, 3, 4, 5, 6, 7, and 8 positions of phenanthrene rings. To determine the structure activity relationship of these derivatives, all 19 isomers were bioassayed with Salmonella tester strains. NAphs substituted at the 4, 6, 7 and 8 positions were mutagenic for TA98, and 1-, 2-, and 3-N-9-AphOs, 6-N-1-AphO and 6-N-4-AphO were mutagenic for TA98 and TA100 without the S9 mix, while 5-N-1-AphO and 5-N-9-AphO were non- or weakly mutagenic. Nitrated derivatives, 6-N-4-Aph, 6-N-9-Aph, 6-N-1-AphO, and 6-N 4-AphO, were powerful mutagens for TA98 and TA100. Mutagenicity was enhanced by mutant strains producing nitroreductase, such as YG1021 and 1026, and by those producing O-acetyltransferase, such as YG1024 and 1029. Nitro derivatives substituted at positions 4 and 5 in the phenanthrene rings were perpendicular, while those at positions 2, 3, 6 and 7 were coplanar to the phenanthrene rings. NAphs substituted at the 1 and 8 positions were noncoplanar due to steric hindrance of the aromatic proton at the peri position. On the other hand, 1,5- and 1,8-dinitro-4-azaphenanthrenes showed high mutagenicity for strains TA98 and TA100 in the absence of the S9 mix, and were strongly enhanced by nitroreductase and O-acetyltransferase, over-producing mutants. Therefore, it was found that the mutagenic potency of NAphs and NAphOs was closely associated with the chemical properties and orientation of nitro substitution of aromatic rings. PMID- 12902150 TI - La(3+), Gd(3+) and Yb(3+) induced changes in mitochondrial structure, membrane permeability, cytochrome c release and intracellular ROS level. AB - Lanthanides (Ln) were known to induce cell apoptosis, which might be the results of their effects on mitochondria (MT). This study was trying to clarify the role of MT and reactive oxygen species (ROS) in Ln-induced apoptosis. We found that micromolar or lower concentration of La(3+), Gd(3+) and Yb(3+) bound to MT and induced swelling of isolated MT; EGTA treatment can inhibit the process. In addition, La(3+), Gd(3+) and Yb(3+) increased the MT membrane fluidity and decreased the MT membrane potential (DeltaPsi(m)). All these were inferred to the results of MT permeability transition pore opening. Release of cytochrome c (Cyt c) from the MT upon incubation with Ln ions was monitored by immunocytochemistry, however, Cyt-c release was observed only in the cytosol of cells. In parallel with these events, there was a higher level of ROS found in the cells exposed to Ln. It was proposed that Ln-induced apoptosis via the MT pathways and it was highly possible that ROS were involved in the mechanism. PMID- 12902151 TI - [35S]-labeling of the Salmonella typhimurium glutathione pool to assess glutathione-mediated DNA binding by 1,2-dibromoethane. AB - Biotransformation of drugs and environmental chemicals to reactive intermediates is often studied with the use of radiolabeled compounds that are synthesized by expensive and technically difficult procedures. In general, glutathione (GSH) conjugation serves as a detoxification mechanism, and conjugation of reactive intermediates with GSH is often a surrogate marker of reactive species formation. However, several halogenated alkanes can be bioactivated by GSH to yield highly reactive GSH conjugates, some of which are DNA-reactive (e.g. conjugates of 1,2 dibromoethane). The purpose of this study was to metabolically radiolabel the in vivo GSH pool of Salmonella typhimurium with a [35S]-label and to examine the GSH mediated bioactivation of a model haloalkane, 1,2-dibromoethane, by measuring the binding of [35S]-label to DNA. The strain of Salmonella used in this study had been transformed previously with the gene that codes for rat glutathione transferase theta 1-1 (GSTT1-1), an enzyme that can catalyze formation of genotoxic GSH conjugates. Bacteria were grown to mid-log phase and then incubated with [35S]-L-cysteine in minimal medium (thio-free) until stationary phase of growth was reached. At this stage, the specific activity of Salmonella GSH was estimated to be 7.1 mCi/mmol by derivatization and subsequent HPLC analysis, and GSTT1-1 enzyme activity was still demonstrable in Salmonella cytosol following growth in a minimal medium. The [35S]-labeled bacteria were then exposed to 1,2 dibromoethane (1 mM), and the Salmonella DNA was subsequently purified to quantify [35S]-binding to DNA. The amount of [35S]-label that was covalently bound to DNA in the GSTT1-1-expressing Salmonella strain (33.2 nmol/mg DNA) was sevenfold greater than that of the control strain that does not express GSTT1-1. Neutral thermal hydrolysis of the DNA yielded a single [35S]-labeled adduct with a similar t(R) as S-[2-(N(7)-guanyl)ethyl]GSH, following HPLC analysis of the hydrolysate. This adduct accounted for 95% of the total [35S]-label bound to DNA. Thus, this [35S]-radiolabeling protocol may prove useful for studying the DNA reactivity of GSH conjugates of other halogenated alkanes in a cellular context that maintains GSH at normal physiological levels. This is also, to our knowledge, the first demonstration of de novo incorporation of [35S]-L-cysteine into the bacterial GSH pool. PMID- 12902152 TI - The effects of genetic variation in N-acetyltransferases on 4-aminobiphenyl genotoxicity in mouse liver. AB - Inbred, congenic and transgenic strains of mice were characterized for acetylation of p-aminobenzoic (PABA) and the carcinogen 4-aminobiphenyl (4ABP). C57Bl/6 mice have the NAT2*8 allele, A/J mice have NAT2*9 and congenic B6.A mice have NAT2*9 on the C57Bl/6 background. The first transgenic strain with human NAT1, the functional equivalent of murine NAT2, was also tested. The murine NAT2*9 allele correlated with a slow phenotype measured with the murine NAT2 selective substrate PABA. The two strains having this allele also had a lower capacity to acetylate 4ABP. A line with five copies of the human NAT1 transgene was bred for at least five generations with either C57Bl/6 or A/J mice. There was no significant change in PABA NAT activity on the C57Bl/6 background but a 2.5 fold increase was seen in hNAT1:A/J compared with A/J. The effect of variation in NATs on 4ABP genotoxicity was assessed in these strains. Twenty-four hours after exposure to a single oral dose of 120 mg 4ABP/kg, hepatic 4ABP-DNA adducts were detected by immunofluoresence in all strains. Nuclear fluorescence intensities (mean+/-S.D.) were 41.1+/-3.6 for C57Bl/6, 37.9+/-1.11 for A/J and 36.3+/-2.44 for B6.A. There was no correlation between murine NAT2 alleles and 4ABP-DNA adduct levels. Similar results were seen with the transgenic strains. The data indicate that the range of variation present in these strains of mice was insufficient to alter susceptibility to 4ABP genotoxicity. The impact of these relatively modest differences in the acetylation of the activation of 4ABP may be masked by other competing biotransformation reactions since 4ABP is a substrate for both NAT1 and NAT2. Mouse models with variation in both isoforms are needed to adequately assess the role of variation in NATs in susceptibility to 4ABP genotoxicity. PMID- 12902153 TI - Sphinganine to sphingosine ratio and predictive biochemical markers of fumonisin B1 exposure in ducks. AB - The kinetics of free sphinganine (Sa), sphinganine to sphingosine ratio (Sa/So), proteins, cholesterol, alanine aminotransferase (ALAT) and lactate dehydrogenase (LDH) were investigated in the course of fumonisin B1 (FB1) exposure in ducks (20 growing males divided into four groups of 5 receiving, respectively, a daily dose of 0, 5, 15 or 45 mg/kg FB1 via oral administration over 12 days). Descriptive statistics of these parameters were also studied in a large number of ducks not exposed to mycotoxins and free of known pathology. Although the toxin at the end of the treatment affected all the parameters investigated, only 2 days of treatment appeared necessary to increase free Sa concentrations in serum, whereas 6 days were necessary to detect a significant effect on Sa/So ratio. Significant differences between control and treated ducks were observed after 4 days of treatment for ALAT and LDH and after 6 and 8 days for cholesterol and proteins concentrations. The minimum doses of FB1 required to determine an effect were assessed using three different methods. This approach reveals that FB1 has greater effects when it is ingested at a low dose for a long time than when ingested at a high dose for a short time. Although the minimum toxic dose of FB1 in ducks remains to be determined, this result must be considered in the context of chronic exposure to the toxin, not only in avian populations. PMID- 12902154 TI - Alterations in human red blood cell membrane properties induced by the lipopolysaccharide from Proteus mirabilis S1959. AB - The effect of lipopolysaccharide (LPS, endotoxin), isolated from Proteus mirabilis S1959 strain, on red blood cell (RBC) membranes in whole cells as well as on isolated membranes was studied. Lipid membrane fluidity, conformational state of membrane proteins and the osmotic fragility of RBCs were examined using electron paramagnetic resonance spectroscopy and spectrophotometric method. Lipid membrane fluidity was determined using three spin-labeled fatty acids: 5-, 12- and 16-doxylstearic acid (5-, 12- and 16-DS). The addition of LPS S1959 to RBC suspension resulted in an increase in membrane fluidity, as indicated by 12-DS. At the concentrations of 0.5 and 1 mg/ml, LPS treatment led to a significant (P<0.05) increase in lipid membrane fluidity in the deeper region of lipid bilayer (determined by 12-DS). The conformational changes in membrane proteins were determined using two covalently bound spin labels, 4-maleimido-2,2,6,6 tetramethylpiperidine-1-oxyl and 4-iodoacetamido-2,2,6,6-tetramethylpiperidine-1 oxyl (ISL). The highest concentration of endotoxin significantly (P<0.05) decreased the relative rotational correlation time of ISL and significantly (P<0.05) increased the osmotic fragility of RBCs. The effect of endotoxin was much more profound in isolated membranes than in intact cells treated with LPS. At the concentrations 0.5 and 1 mg/ml, LPS led to a significant increase in h(w)/h(s) ratio. These results indicated increased membrane protein mobility, mainly in the spectrin-actin complex in membrane cytoskeleton. These data suggest that LPS-induced alterations in membrane lipids and cytoskeleton proteins of RBCs lead to loss of membrane integrity. PMID- 12902155 TI - Detection of benzylic adducts in DNA and nucleotides from 7-sulfooxymethyl-12 methylbenz[a]anthracene and related compounds by 32P-postlabeling using new TLC systems. AB - 7,12-Dimethylbenz[a]anthracene (DMBA) is a highly potent experimental carcinogen, that must be transformed to its ultimate carcinogenic form in vivo. The meso region theory of aromatic hydrocarbon carcinogenesis predicts that 7 hydroxymethyl sulfate (7-HMBA) ester plays a major role in the metabolic activation, benzylic DNA adduct formation and complete carcinogenicity of HMBA and DMBA. This study was undertaken to detect highly lipophilic benzylic DNA adducts resulting from the reaction between 7-hydroxymethy sulfate ester of HMBA (7-SMBA) and DNA as well as determine their DNA base selectivity. Synthetic 7 SMBA was incubated with DNA (800 microg/ml) and individual deoxynucleoside 3' monophosphates (600 microg/ml) and benzylic adducts were analyzed by 32P postlabeling/TLC following their enrichment with butanol extraction. Dilute ammonium hydroxide-based solvents were developed to detect the highly lipophilic aralkyl adducts. The reaction with DNA, dGp and dAp gave rise to multiple adducts; dCp and dTp showed no significant adducts. Chromatographic comparison revealed that the major DNA adduct was derived from dG. The methodology developed was also found applicable for highly lipophilic adducts resulting from sulfate esters of structurally-related metabolites of DMBA. PMID- 12902156 TI - Involvement of intracellular glutathione in zinc deficiency-induced activation of hepatic stellate cells. AB - Hepatic stellate cells (HSC) play an important role in the development of liver cirrhosis. They are a major source of extracellular matrix and during fibrogenesis undergo an activation process characterized by increased proliferation and collagen synthesis. In this study, we investigated the anti fibrogenic effect of zinc supplementation on zinc deficiency induced HSC activation. Isolated HSC were incubated with or without zinc chelator, diethylenetriamine penta-acetic acid (DTPA). Type I collagen expression in HSC was detected by immunohistochemistry. The involvement of glutathione (GSH) homeostasis in the anti-fibrogenic action of zinc was also investigated, as GSH is implicated in many cellular events, such as regulation of cell proliferation, remodeling of extracellular matrix and oxidative stress. Intracellular GSH was measured by HPLC. Enhanced type I collagen expression, apoptosis and cell cycle arrest were found in HSC when DTPA was added, but they were inhibited with supplementation with zinc. Zinc deficiency caused a reduction in intracellular GSH 8 h after the addition of DTPA compared with control levels. The results of this study show that in HSC, the chelation of zinc triggers a progression of collagen synthesis and this involves the depletion of intracellular GSH levels after the addition of DTPA. PMID- 12902157 TI - Moonlighting proteins: old proteins learning new tricks. AB - Recently, several laboratories identifying proteins involved in the complex processes of replication, transcription and tumor suppression found that the 'new' protein they discovered had another, previously identified, function. A single protein with multiple functions might seem surprising, but there are actually many cases of proteins that 'moonlight', or have more than one role in an organism. As well as adding to the number and types of proteins that are known to moonlight, these new examples add to our understanding of the potential importance of moonlighting proteins. PMID- 12902158 TI - Evolution of cis-regulatory elements in duplicated genes of yeast. AB - An increasing number of studies report that functional divergence in duplicated genes is accompanied by gene expression changes, although the evolutionary mechanism behind this process remains unclear. Our genomic analysis on the yeast Saccharomyces cerevisiae shows that the number of shared regulatory motifs in the duplicates decreases with evolutionary time, whereas the total number of regulatory motifs remains unchanged. Moreover, genes with numerous paralogs in the yeast genome do not have especially low number of regulatory motifs. These findings indicate that degenerative complementation is not the sole mechanism behind expression divergence in yeast. Moreover, we found some evidence for the action of positive selection on cis-regulatory motifs after gene duplication. These results suggest that the evolution of functional novelty has a substantial role in yeast duplicate gene evolution. PMID- 12902159 TI - Genomic analysis of gene expression relationships in transcriptional regulatory networks. AB - From merging several data sources, we created an extensive map of the transcriptional regulatory network in Saccharomyces cerevisiae, comprising 7419 interactions connecting 180 transcription factors (TFs) with their target genes. We integrated this network with gene-expression data, relating the expression profiles of TFs and target genes. We found that genes targeted by the same TF tend to be co-expressed, with the degree of co-expression increasing if genes share more than one TF. Moreover, shared targets of a TF tend to have similar cellular functions. By contrast, the expression relationships between the TFs and their targets are much more complicated, often exhibiting time-shifted or inverted behavior. Further information is available at http://bioinfo.mbb.yale.edu/regulation/TIG/ PMID- 12902160 TI - Control of cross-over by single-strand DNA resection. AB - Control of DNA cross-overs is necessary for meiotic recombination and genome integrity. The frequency of cross-overs is dependent on homology length and the conversion tract, but the mechanisms underlying the regulation of cross-overs remain unknown. We propose that 5'-end resection, a key intermediate in double strand break repair, could determine the formation of cross-overs. Extensive DNA resection might favor gene conversion without cross-over by channeling recombination events through synthesis-dependent strand-annealing. In reactions with short regions of homology, resection beyond the homologous sequence would impede Holliday junction formation and, consequently, cross-over. Extensive DNA resection could be an effective mechanism to prevent reciprocal exchanges between dispersed DNA sequences, and thus contribute to the genome stability. PMID- 12902161 TI - A stain upon the silence: genes escaping X inactivation. AB - X-chromosome inactivation is a remarkable epigenetic event in mammalian females that results in the transcriptional silencing of one of the pair of X chromosomes. However, not all X-linked genes are subject to inactivation, and in humans, the proportion of genes on the X chromosome that escapes inactivation is more than 15%. Here we examine the causes and consequences of failure to silence the entire X chromosome. We discuss the impact of the evolutionary history of the X (and Y) chromosome, and the bioinformatic approaches that promise to provide new insights into the genomic architecture of genes or regions that escape X chromosome inactivation. PMID- 12902162 TI - Telomere maintenance and DNA replication: how closely are these two connected? AB - The maintenance of the DNA at chromosome ends, the telomeres, depends on conventional semiconservative replication and on the action of telomerase, a specialized reverse transcriptase. Current research strongly suggests a regulatory interplay between this conventional semiconservative replication and telomerase, thus ensuring that no sequences are lost at the very ends of the telomeres during replication. Here, we describe recent findings on the interactions between the conventional replication machinery and telomere replication, and we discuss how DNA-integrity checkpoints might impinge on both the processing of the telomeric DNA ends and the establishment of the DNA end structure required for end protection and genome stability. PMID- 12902163 TI - Maneuvering for advantage: the genetics of mouse susceptibility to virus infection. AB - Genetic studies of host susceptibility to infection contribute to our understanding of an organism's response to pathogens at the immunological, cellular, and molecular levels. In this review we describe how the study of host genetics in mouse models has helped our understanding of host defense mechanisms against viral infection, and how this knowledge can be extended to human infections. We focus especially on the innate mechanisms that function as the host's first line of defense against infection. We also discuss the main issues that confront this field, as well as its future. PMID- 12902164 TI - Transcriptional mechanisms in osteoblast differentiation and bone formation. AB - Osteoblasts, the cells responsible for bone formation, differentiate from mesenchymal cells. Here, we discuss transcription factors that are involved in regulating the multistep molecular pathway of osteoblast differentiation. Runx2 and Osx, a newly identified zinc-finger-containing protein, are transcription factors that are expressed selectively and at high levels in osteoblasts. Null mutations of either leads to a complete absence of bone in mice. Runx2 plus its companion subunit Cbf beta are needed for an early step in this pathway, whereas Osx is required for a subsequent step, namely the differentiation of preosteoblasts into fully functioning osteoblasts. The finding that Osx-null cells acquire a chondrocyte phenotype implies that Osx is a negative regulator of Sox9 and of the chondrocyte phenotype. This leads to the hypothesis that Osx might have a role in the segregation of osteoblasts from osteochondroprogenitors. We also discuss recent progress in studies of other transcription factors that affect skeletal patterning and development. PMID- 12902165 TI - Star Trek replicators and diatom nanotechnology. AB - Diatoms are single celled algae, the 10(5)-10(6) species of which create a wide variety of three-dimensional amorphous silica shells. If we could get them to produce useful structures, perhaps by compustat selection experiments (i.e. forced evolution of development or evodevo), their exponential growth in suspension cultures could compete with the lithography techniques of present day nanotechnology, which have limited 3D capabilities. Alternatively, their fine detail could be used for templates for MEMS (micro electro mechanical systems), or their silica deposition systems isolated for guiding silica deposition. A recent paper has demonstrated that silica can be replaced atom for atom without change of shape--a step towards the Star Trek replicator. PMID- 12902166 TI - Putting some spine into alternative splicing. AB - Spinal muscular atrophy is a neurodegenerative disease caused by mutations of the SMN1 gene. The homologous SMN2 gene is unable to complement SMN1 because of a crucial mutation in an exonic splicing enhancer, leading to alternative splicing and exclusion of exon 7. Two recent papers show that the defect in splicing of exon 7 of SMN2 is specifically corrected by small synthetic effectors. These new and specific approaches have potential in the treatment of diseases caused by defective splicing. PMID- 12902167 TI - Organelle identification and proteomics in plant cells. PMID- 12902169 TI - Will genomics revolutionise pharmaceutical R&D? AB - The successful identification of drug targets requires an understanding of the high-level functional interactions between the key components of cells, organs and systems, and how these interactions change in disease states. This information does not reside in the genome, or in the individual proteins that genes code for, it is to be found at a higher level. Genomics will succeed in revolutionising pharmaceutical research and development only if these interactions are also understood by determining the logic of healthy and diseased states. The rapid growth in biological databases, models of cells, tissues and organs, and in computing power has made it possible to explore functionality all the way from the level of genes to whole organs and systems. Combined with genomic and proteomic data, in silico simulation technology is set to transform all stages of drug discovery and development. The major obstacle to achieving this will be obtaining the relevant experimental data at levels higher than genomics and proteomics. PMID- 12902170 TI - What can spores do for us? AB - Many organisms have the ability to form spores, a remarkable phase in their life cycles. Compared with vegetative cells, spores have several advantages (e.g. resistance to toxic compounds, temperature, desiccation and radiation) making them well suited to various applications. The applications of spores that first spring to mind are bio-warfare and the related, but more positive, field of biological control. Although they are often considered metabolically inert, spores can also be used as biocatalysts. Other uses for spores are found in the fields of probiotics, tumour detection and treatment, biosensing and in the "war against drugs". PMID- 12902171 TI - The relevance of alternative RNA splicing to pharmacogenomics. AB - The importance of alternative RNA splicing in the generation of genetic diversity is now widely accepted. This article highlights how alternative RNA splicing can have an impact on drug efficacy and safety, and demonstrates its potential pharmacogenomic value. The analysis of the repertoire of alternative RNA splicing events could potentially identify markers of pharmacogenomic relevance with high sensitivity and specificity and also provides a route through which genes can be selected for single nucleotide polymorphism (SNP) genotyping. Recent methodological advances, including microarray and splice-dedicated expression profiling, have made it possible to perform high-throughput alternative splicing analyses. PMID- 12902172 TI - Teaching cells new tricks. AB - The direct conversion of one differentiated cell type into another--a process referred to as transdifferentiation--would be beneficial for producing isogenic (patient's own) cells to replace sick or damaged cells or tissue. Adult stem cells display a broader differentiation potential than anticipated and might contribute to tissues other than those in which they reside. As such, they could be worthy therapeutic agents. Recent advances in transdifferentiation involve nuclear transplantation, manipulation of cell culture conditions, induction of ectopic gene expression and uptake of molecules from cellular extracts. These approaches open the doors to new avenues for engineering isogenic replacement cells. To avoid unpredictable tissue transformation, nuclear reprogramming requires controlled and heritable epigenetic modifications. Considerable efforts remain to unravel the molecular processes underlying nuclear reprogramming and evaluate stable of the changes in reprogrammed cells. PMID- 12902173 TI - Gone gene fishing: how to catch novel marine antimicrobials. AB - Medical or health-promoting products of marine origin are often regarded with skepticism--some, such as shark fins and cod liver oil, are frequently perceived as low-tech "alternative treatments" largely because they have not been exploited to their full potential. The marine environment is an enormous source of biodiversity--80% of all life is found under the oceans' surfaces--yet very little of this rich resource has been utilized. Furthermore, most marine organisms rely heavily on antimicrobial components of their innate immune defenses to combat pathogens. The past three years has seen a revolution in the methods used to identify novel antimicrobials from marine sources; among the most promising are marine cationic antimicrobial peptides (CAPs). PMID- 12902174 TI - One hundred and one Dalmatians -- the importance of being 'patch'? PMID- 12902175 TI - Micronutrient research: why a cow is not a large rat. PMID- 12902176 TI - Gut feelings on the use of ultrasonography as a diagnostic aid. PMID- 12902177 TI - Ultrasonography in gastrointestinal disease in cattle. AB - Ultrasonography is an ideal diagnostic tool for investigating gastrointestinal disorders in cattle. It is performed on standing non-sedated cattle using a 3.5 MHz linear transducer. In animals with traumatic reticuloperitonitis, inflammatory fibrinous changes, and abscesses can be imaged; however, magnets and foreign bodies are difficult to visualize because of the gas content of the reticulum. Ultrasonography can be used to assess the size, position and contents of the abomasum. Percutaneous ultrasound-guided abomasocentesis can be performed to evaluate the nature and chemical composition of its contents. In left displacement of the abomasum, the abomasum is seen between the left abdominal wall and the rumen. It contains fluid ingesta ventrally and a gas cap of varying size dorsally. Occasionally, the abomasal folds are seen in the ingesta. In cattle with right displacement of the abomasum, the liver is displaced medially from the right abdominal wall by the abomasum, which has an ultrasonographic appearance similar to that described for left displacement. Motility and diameter of the intestine are the most important criteria for ultrasonographic assessment of ileus. However, the cause of the ileus is rarely determined using ultrasonography. In cases with ileus of the small intestine, there is at least one region of dilatation of the intestine and motility is reduced or absent. In cattle with caecal dilatation, the caecum can always be imaged from the right lateral abdominal wall. The wall of the caecum closest to the transducer appears as a thick, echogenic, semi-circular line. PMID- 12902178 TI - The role of essential trace elements in embryonic and fetal development in livestock. AB - This review addresses the concept that essential trace minerals play a vital role in many enzymatic and metabolic pathways that are critical for conceptus development during pregnancy in livestock species. The conceptus relies entirely on the maternal system for a sufficient supply of trace minerals and other nutrients needed for normal development. If this supply is inadequate, growth and/or health of the conceptus can be affected adversely, and many of these effects carry over into the neonatal period. Information, accumulated in our laboratory and presented herein, indicates that zinc, copper and manganese are among the trace minerals that have the greatest impact on reproduction. For example, levels of zinc, copper and manganese were several fold greater in the conceptus than in other reproductive tissues, indicating that the conceptus preferentially accumulates these minerals, an action that may be important for conceptus development, growth and survival. Moreover, some recent results indicate that increasing the biological availability of zinc, copper and manganese, by attachment to short peptide chains (i.e., proteinated trace minerals) can enhance reproductive performance of swine. Mineral concentrations in conceptuses from female pigs consuming proteinated trace minerals were greater than those from females that consumed only inorganic mineral salts. Elucidating the mechanisms whereby conceptus development and survival are enhanced by essential trace minerals may lead to development of specific feeding programs to increase the number and health of offspring at parturition, thereby allowing for further improvements in production efficiency in animal agriculture. PMID- 12902179 TI - Apoptosis and the loss of chondrocyte survival signals contribute to articular cartilage degradation in osteoarthritis. AB - Apoptotic death of articular chondrocytes has been implicated in the pathogenesis of osteoarthritis (OA). Apoptotic pathways in chondrocytes are multi-faceted, although some cascades appear to play a greater in vivo role than others. Various catabolic processes are linked to apoptosis in OA cartilage, contributing to the reduction in cartilage integrity. Recent studies suggest that beta1-integrin mediated cell-matrix interactions provide survival signals for chondrocytes. The loss of such interactions and the inability to respond to IGF-1 stimulation may be partly responsible for the hypocellularity and matrix degradation that characterises OA. Here we have reviewed the literature in this area of cartilage cell biology in an effort to consolidate the existing information into a plausible hypothesis regarding the involvement of apoptosis in the pathogenesis of OA. Understanding of the interactions that promote chondrocyte apoptosis and cartilage hypocellularity is essential for developing appropriately targeted therapies for inhibition of chondrocyte apoptosis and the treatment of OA. PMID- 12902180 TI - Axial compression generated by cortical and cancellous lag screws in the equine distal phalanx. AB - Lag screw fixation using single 4.5 mm cortical bone screws is a recommended technique for repair of mid-sagittal plane fractures of the distal phalanx in adult horses. However, implant infection and technical difficulties in obtaining adequate interfragmentary compression have made this surgical procedure somewhat controversial. We hypothesized that use of larger diameter screws would result in increased axial compression and improved stability of this fracture. Paired distal phalanges from the forelimbs of 10 adult horses were collected at necropsy and divided in half in the midsagittal plane. Using a randomized block study design, four types of bone screws (4.5 mm cortical, 5.5 mm cortical, 6.5 mm cancellous pre-tapped, and 6.5 mm cancellous non-tapped) were inserted to a depth of 15 mm. During screw insertion, the axial force generated under the screw head was measured with a load washer containing a piezoelectric force transducer, while torque of insertion was recorded with a torsional testing machine. The 6.5mm screw inserted after pre-tapping generated significantly greater axial force (2781 N) than the 4.5 mm (1522 N), 5.5 mm (2073 N) or 6.5 mm non-tapped (2295 N) screws. The relationship between maximal applied torque and axial force generated was linear for each screw type. Each unit of torque applied during insertion of cortical screws resulted in a greater increase in axial compression, as compared to cancellous screws. These data suggest that use of larger diameter screws would result in improved interfragmentary compression of distal phalangeal fractures. PMID- 12902181 TI - Analysis of systematic effects on congenital sensorineural deafness in German Dalmatian dogs. AB - We have analysed the systematic influences, phenotypic colour markers and the additive genetic variation for congenital sensorineural deafness (CSD) in German Dalmatian dogs in order to help elucidate the importance of phenotypic breed characteristics for genetic differences of CSD. Linear animal models using restricted maximum likelihood methods were employed to estimate variance components. Data were obtained from all three German Dalmatian kennel clubs associated with the German Association for Dog Breeding and Husbandry (VDH). CSD was recorded by standardized protocols for brainstem auditory-evoked response (BAER). The material included 1899 German Dalmatian dogs from 354 litters in 169 different kennels. BAER testing results were from the years 1986 to 1999. Pedigree information was available for up to seven generations. The animal model regarded the fixed effects of sex, coat colour, eye colour, presence of patches, litter size, percentage of examined puppies per litter, kennel club, and inbreeding coefficient. The common environment of the litter and kennel as well as the additive genetic effect of the animal were taken into account as randomly distributed effects. The fixed effects of eye colour, percentage of puppies examined per litter and kennel club were significant in the mixed model analysis. A significant proportion of additive genetic variation could be shown despite corrections for phenotypic colour variants. The heritability estimate for CSD in German Dalmatian dogs was h(2)=0.27+/-0.07. The additive genetic correlation of CSD with presence of blue eyes was r(g)=0.53+/-0.41 and with presence of patches r(g)=-0.36+/-0.24. We concluded that additional genes other than those associated with phenotypic colour markers in German Dalmatian dogs significantly contribute to the occurrence of CSD. PMID- 12902182 TI - Pharmacodynamics and pharmacokinetics of cefoperazone and cefamandole in dogs following single dose intravenous and intramuscular administration. AB - The pharmacokinetics and intramuscular (i.m.) bioavailability of cefoperazone and cefamandole (20mg/kg) were investigated in dogs and the findings related to minimal inhibitory concentrations (MICs) for 90 bacterial strains isolated clinically from dogs. The MICs of cefamandole for Staphylococcus intermedius (MIC(90) 0.125 microg/mL) were lower than those of cefoperazone (MIC(90) 0.5 micro/mL) although the latter was more effective against Escherichia coli strains (MIC(90) 2.0 microg/mL vs. 4.0 microg/mL). The pharmacokinetics of the drugs after intravenous administrations were similar: a rapid distribution phase was followed by a slower elimination phase (t((1/2)lambda2) 84.0+/-21.3 min for cefoperazone and 81.4+/-9.7 min for cefamandole). The apparent volume of distribution and body clearance were 0.233 L/kg and 1.96 mL/kg/min for cefoperazone, 0.190 L/kg and 1.76 mL/kg/min for cefamandole. After i.m. administration the bioavailability and peak serum concentration of cefamandole (85.1+/-13.5% and 35.9+/-5.4 microg/mL) were significantly higher than cefoperazone (41.4+/-7.1% and 24.5+/-3.0 micog/mL), but not the serum half-lives (t(1/2el) 134.3+/-12.6 min for cefoperazone and 145.4+/-12.3 min for cefamandole). The time above MIC(90) indicated that cefamandole can be administered once daily to dogs for the treatment of staphylococcal infections (T>MIC for S. intermedius 23.8+/-0.3 and for Staphylococcus aureus 21.6+/-0.6h). PMID- 12902183 TI - Experimental infection of brushtail possums (Trichosurus vulpecula) with Mycobacterium bovis by conjunctival instillation. AB - In New Zealand, the brushtail possum (Trichosurus vulpecula) is the major wildlife reservoir of Mycobacterium bovis. Procedures for experimentally infecting possums are required to study the pathogenesis of the disease and to challenge possums in vaccine efficacy studies. Conjunctival instillation of a suspension of M. bovis was effective in producing bovine tuberculosis in captive possums. The experimental disease progressed slowly with the development of palpable lesions in superficial lymph node lesions, both characteristics of the disease in wild, naturally infected possums. At necropsy there was widespread distribution of macroscopic and microscopic lesions. The proportion of possums that became diseased, the rate of development and severity of lesions, the severity of clinical signs, all increased when the dose of M. bovis was increased. Of the three doses used, the medium dose (1000-2000 colony forming units) produced the disease with the most desired characteristics. As a procedure for exposing possums to infection with M. bovis the conjunctival route has advantages in that it is simple and safe to perform, and possums need only to be sedated for infection. PMID- 12902184 TI - Identification of a bisegmented double-stranded RNA virus (picobirnavirus) in calf faeces. AB - To determine the incidence of rotavirus infection among dairy herds in the State of Sao Paulo, Brazil, 576 faecal samples obtained from calves aged 1-45 days with and without diarrhoea, reared on 63 dairy cattle farms, were analyzed. Polyacrylamide gel electrophoresis (PAGE) identified 28 samples positive for group A rotavirus, while four samples, two diarrhoeic and two non-diarrhoeic, showed a bisegmented genome with a typical picobirnavirus pattern. Electron microscopy revealed spherical virus particles with a diameter of 37 nm and without a defined surface structure. The present study is the first report of a bisegmented virus identified in cattle in Brazil. PMID- 12902185 TI - Transcutaneous ultrasound-guided pancreatic biopsy in cattle and its safety: a preliminary report. AB - This study describes a free-hand technique for percutaneous pancreatic biopsy in cattle with ultrasound-guidance using a 14G spinal biopsy needle. Its safety was evaluated based on 36 consecutive procedures. To assess the immediate effects of pancreatic biopsy, 31 cows were necropsied shortly after the procedure and examined. The remaining five cows were examined daily for eight days and then necropsied and examined. No life-threatening complications nor clinically detectable abnormalities were observed. Changes indicative of inflammation were not apparent in total and differential WBC counts or in total protein and fibrinogen concentrations. A small increase of amylase activity was detected in only one cow kept for the eight-day observation period. Serum lipase activity increased significantly on day four after biopsy. Urea nitrogen, creatinine, glucose and the activities of aspartate aminotransferase, alanine aminotransferase, and gamma-glutamyltransferase remained within reference ranges. Changes in the peritoneum and pancreas observed at necropsy were negligible. We conclude that percutaneous ultrasound-guided pancreatic biopsy did not appear to influence the cow's condition adversely and the procedure provided an excellent method of obtaining a pancreatic specimen for histological examination. The procedure was considered safe, fast, cost-effective, and practical when performed properly. We believe that the technique can be used in cows with suspected pancreatic disease for making an ante mortem diagnosis. PMID- 12902186 TI - Antagonistic effects of intravenous or epidural atipamezole on xylazine-induced dorsolumbar epidural analgesia in cattle. AB - This study was performed to clarify the antagonistic actions of intravenous or epidural atipamezole on the sedative and analgesic effects of xylazine administered between the epidural fat and dura mater through the first interlumbar space in cattle. Cattle received 5 mL of a solution containing 0.05 mg x kg(-1) xylazine in 0.9% saline. Thirty minutes later, 5 mL of 0.9% saline was administered through the same needle (treatment 1) (XSE). In treatments 2 (XAE) and 3 (XAV), 5 mL of a solution containing 0.025 mg x kg(-1) atipamezole in 0.9% saline was administered epidurally or intravenously, respectively. Sedation and analgesia were similar in all three treatment groups and could be reversed by atipamezole given by either route. In the XAV treatment, the flank area relapsed into analgesia 25+/-5.8 min following reversal of the analgesic effect, and was maintained for 112.5+/-63.8 min. The present study confirmed that the sedative and analgesic effects of xylazine are completely reversed by atipamezole and can be influenced by the epidural fat in cattle. Furthermore, it seems probable that analgesia following epidural administration of xylazine is mediated by alpha(2) adrenergic receptors, not by a local anaesthetic effect. PMID- 12902187 TI - Transplacental transfer of propofol in pregnant ewes. AB - Propofol is an injectable anaesthetic that is currently used both in veterinary and human medicine for the induction and maintenance of anaesthesia. Although little is known about the pharmacokinetics of propofol in fetuses, it is widely used in obstetric procedures, particularly in caesarean section. This study determines the pharmacokinetics of propofol in pregnant ewes in the last third of pregnancy, and placental transfer and pharmacokinetics in fetuses after the administration of a 6 mg/kg intravenous (i.v.) bolus (phase 1) or a 6 mg/kg i.v. bolus followed by continued infusion of 0.4 mg/kg/min. In ewes, the area under the blood concentration-time curve (AUC) and C(max) (8.6 mgh/mL and 9.5mg/mL, respectively) was higher than those of the fetus (1.6 mgh/mL and 1.19 mg/mL, respectively). The mean half-life was 0.5h in the dam and 1.1h in the fetus. PMID- 12902188 TI - Observations on aggregated lymphoid nodules in the cardiac glandular areas of the Bactrian camel (Camelus bactrianus). AB - Aggregated lymphoid nodules are an important part of the gut-associated lymphoid tissue (GALT). They are mainly distributed in the ileum and appendix of animals and humans but their distribution in the cardiac glandular area has not been reported. A study of stomach histology in the Bactrian camel has revealed that the nodules are distributed as a band-like region along the ventral wall of the stomach neck, at the beginning of the cranial enlargement and on the lesser curvature. The mucous folds are thicker in these regions than where there are no aggregated lymphoid nodules. The nodules appeared similar to ileal aggregated lymphoid nodules found in other animals and consisted of typical polymorphological lymphatic nodules arranged in a single continuous row occupying the submucosa and forming mucosal folds together with the mucous membrane. The whole mucous membrane with cardiac glands, diffuse lymphatic tissue and solitary lymphoid nodules in the lamina propria were found to cover the aggregated lymphoid nodule regions, but some nodules with a typical corona extended into the lamina propria and were covered with follicle-associated epithelium devoid of cardiac glands. These findings indicate that the stomach of the Bactrian camel possesses not only a special structure of digestion but also has characteristic immunological morphology. PMID- 12902189 TI - Presence of focal and multifocal paratuberculosis lesions in mesenteric lymph nodes and the ileocaecal valve of cattle positive to the tuberculin skin test. PMID- 12902190 TI - The organochlorine o,p'-DDD disrupts the adrenal steroidogenic signaling pathway in rainbow trout (Oncorhynchus mykiss). AB - The mechanisms of action of o,p'-DDD on adrenal steroidogenesis were investigated in vitro in rainbow trout (Oncorhynchus mykiss). Acute exposures to o,p'-DDD inhibited ACTH-stimulated cortisol secretion while cell viability decreased significantly only at the highest concentration tested (200 microM o,p'-DDD). Stimulation of cortisol secretion with a cAMP analogue (dibutyryl-cAMP) was inhibited at a higher concentration than that needed to inhibit ACTH-stimulated cortisol synthesis in cells exposed to o,p'-DDD. Forskolin-stimulated cortisol secretion and cAMP production, and NaF-stimulated cAMP production were inhibited in a concentration-dependent manner by o,p'-DDD. In contrast, basal cortisol secretion was stimulated while basal cAMP production was unaffected by o,p'-DDD. Pregnenolone-stimulated cortisol secretion was enhanced by o,p'-DDD at a physiologically relevant pregnenolone concentration, while o,p'-DDD inhibited cortisol secretion when a pharmacological concentration of pregnenolone was used. Our results suggest that the cAMP generation step is a target in o,p'-DDD mediated disruption of ACTH-stimulated adrenal steroidogenesis in rainbow trout but that other downstream targets such as steroidogenic enzymes responsible for cortisol synthesis might also be affected. PMID- 12902191 TI - Pharmacokinetics of ethylene in man by on-line laser photoacoustic detection. AB - The pharmacokinetics of ethylene are determined using laser-based photoacoustic detection and a closed chamber setup. Concentration-time data are analyzed using a two-compartment and a physiologically based pharmacokinetic (PBPK) model. Endogenous production was 92 +/- 13 pmol/h/kg for the two-compartment model and 75 +/- 10 pmol/h/kg for the PBPK model. These values agree with previous work at our department but are significantly lower than published values based on gas chromatography. The blood:air partition coefficient in the PBPK model was determined by curve fitting, because simulations based on published values did not agree well with data. Curve fitting gave a value of 0.092 +/- 0.029. The real time nature and high sensitivity of photoacoustic detection make it a useful addition to gas chromatography in closed chamber studies. PMID- 12902192 TI - Selective activation of Src family kinases and JNK by low levels of chromium(VI). AB - Inhaled hexavalent chromium (Cr(VI)) promotes pulmonary disease and lung cancer through poorly defined mechanisms. These mechanisms were studied in A549 lung epithelial cells to investigate the hypothesis that nontoxic Cr(VI) exposures selectively activate cell signaling that shifts the balance of gene transcription. These studies demonstrated that nontoxic doses of Cr(VI) (10 microM) increased reactive oxygen species and selectively activated c-Jun N terminal kinase (JNK), relative to ERK or p38 MAP kinase. In contrast, only toxic, nonselective levels of exogenous oxidants stimulated JNK. However, JNK activation in response to Cr(VI) and exogenous H(2)O(2) (1 mM) shared requirements for intracellular thiol oxidation, activation of Src family kinases, and p130(cas) (Cas). Cr(VI) did not mimic H(2)O(2)-mediated stimulation of JNK in fibroblasts containing only Src and did not activate Src or Yes in A549 cells. Instead, Fyn and Lck were activated in A549 cells, indicating activation of specific Src family kinases in response to Cr(VI). Finally, Cr(VI) was demonstrated to directly activate purified Fyn in vitro and the majority of this activation did not require oxidant generation. These data suggest that nontoxic levels of Cr(VI), which can shift patterns of gene transcription, are selective in their activation of cell signaling and that Cr(VI) can directly activate Src family kinases independently of reactive oxygen species generation. PMID- 12902193 TI - Formation of 8-oxo-7,8-dihydro-2'-deoxyguanosine in rat lung DNA following subchronic inhalation of carbon black. AB - Chronic high-dose inhalation of carbon black (CB) can produce carcinomas in rat lungs. The mechanisms underlying this response are uncertain. It has been hypothesized that chronic inflammation and cell proliferation may play a role in the development of tumors after high dose, long-term contact of the particles with lung epithelial cells. In this investigation, we analyzed the formation of a known mutagenic lesion [8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxo-dG)] in the lung DNA of rats following subchronic inhalation of CB (Printex-90 and Sterling V). Briefly, female Fischer 344 rats were exposed for 6 h/day, 5 days/week for 13 weeks to 1, 7, and 50 mg/m(3) of Printex-90 (16 nm; specific surface area 300 m(2)/g) and to 50 mg/m(3) of Sterling V CB (70 nm; surface area of 37 m(2)/g). The exposure concentration of Sterling V was selected to be equivalent in terms of retained mass in the lung to the high dose of Printex-90 at the end of exposure. However, in terms of retained particle surface area, the retained lung dose of Sterling V was equivalent to the mid-dose of Printex 90. This design allows comparison of results on the basis of retained particle mass as well as retained particle surface area between the two CB particles. The formation of 8 oxo-dG in the lung DNA was assessed using a reverse phase HPLC system coupled with UV and electrochemical (EC) detection. After 13 weeks of exposure, measurements were made on lung samples obtained at the end of the exposure and a 44-week recovery period in clean air. Lung burdens of CB were determined at both time points as well as differential cell populations from bronchoalveolar lavage fluid (BAL). The results indicate that lung particle overload was achieved after exposure to 7 and 50 mg/m(3) (Printex-90) and 50 mg/m3 (Sterling V) but not at 1 mg/m(3) (Printex-90). Consistent with these results, a significant increase (P < 0.05) in 8-oxo-dG induction was observed following 13 weeks of exposure to 50 mg/m(3) Printex-90 and at 7 and 50 mg/m(3) after the 44-week recovery period. Interestingly, no increase in 8-oxo-dG was observed for Sterling V CB at either time point despite lung particle overload. Although the retained mass dose of Sterling V at the end of exposure was even higher than for Printex 90 (50 mg/m(3) exposure group) (approximately 7.6 vs 4.8 mg), the surface area of the retained Sterling V was similar to that of the retained Printex 90 of the mid-dose exposure (7 mg/m(3)) (approximately 0.2 m(2) in both groups). Since both Sterling V (50 mg/m(3)) and Printex 90 (7 mg/m(3)) did not induce significant increases in 8-oxo-dG in the lung at the end of the 13-week exposure, this finding indicates that a retained large particle mass is not always correlated with similar adverse effects but that particle surface area is a better dose parameter. The lower effect per unit mass dose seen with Sterling V is consistent with earlier studies showing that particle surface area of low toxicity particles is a more appropriate dosemetric for induction of inflammation in the lungs than particle mass (Oberdorster et al., 1994, 2001; Brown et al. 2001; Donaldson et al., 2002). An increase (p < 0.05) in lung lavage neutrophils was observed at 7 mg/m(3) (Printex-90) and 50 mg/m(3) (Printex-90 and Sterling V) at the 13-week exposure period and again at 50 mg/m(3) (Printex-90 and Sterling V, 44-week recovery period). Our current findings suggest that prolonged, high-dose exposure to CB can promote oxidative DNA damage that is consistent with the hypothesis that inflammatory cell-derived oxidants may play a role in the pathogenesis of rat lung tumors following long-term high-dose exposure to CB in rats. PMID- 12902194 TI - Evidence that the lichen-derived scabrosin esters target mitochondrial ATP synthase in P388D1 cells. AB - Scabrosin esters (SEs), which have been recently isolated from the lichen Xanthoparmelia scabrosa, belong to the epipolythiodioxopiperazine (ETP) class of secondary metabolites characterized by possession of a reactive disulfide bond. Colony forming assays show that these toxins are active against human tumor cell lines at nanomolar concentrations. Other members of the ETP class of toxins such as gliotoxin have been shown to induce apoptosis in cells, although the cellular target(s) of the ETP toxins is currently unknown. ETP toxins have been shown to inhibit a variety of enzymes via interaction with sensitive cysteine residues. Here we show that the typical scabrosin ester acetate butyrate induces early mitochondrial membrane hyperpolarization assessed by JC-1 staining accompanied by apoptotic cell death. The toxin lowers ATP in intact cells and inhibits the rate of ATP synthesis in permeabilzed cells. Comparison with the effects of the known ATP synthase inhibitor oligomycin B is consistent with ATP synthase as an early target in scabrosin ester-induced cell death. PMID- 12902195 TI - Effects of several dioxin-like compounds on estrogen metabolism in the malignant MCF-7 and nontumorigenic MCF-10A human mammary epithelial cell lines. AB - In human breast tissue, estrone (E(1)) and estradiol (E(2)) are mainly hydroxylated by cytochrome P450 1A1 (CYP1A1) and 1B1 (CYP1B1) to 2 hydroxyestrogens (2-OHE(1/2)) and 4-hydroxyestrogens (4-OHE(1/2)), respectively. Several studies show that 4-OHE(1/2), but not 2-OHE(1/2), may act as a carcinogen and a high estrogen 4-/2-hydroxylation ratio appears to be a marker for the presence of neoplasms. In this study, we investigated the effects of several dioxin-like compounds on estrogen 2- and 4-hydroxylation in a malignant (MCF-7) and a nontumorigenic (MCF-10A) human mammary epithelial cell line. 2- and 4 methoxyestrogen (MeOE(1/2)) formations were used as measures of the 2- and 4 hydroxylation pathways, respectively. 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD), 2,3,4,7,8-pentachlorodibenzofuran (PCDF), 3,3',4,4',5-pentachlorobiphenyl (PCB 126), and 3,3'4,4',5,5'-hexachlorobiphenyl (PCB 169) concentration dependently induced 2-MeOE(1/2) formation and ethoxyresorufin-O-deethylation (EROD) activity through induced CYP1A1 expression in MCF-7 and MCF-10A cells. 2,3',4,4',5 pentachlorobiphenyl (PCB 118) had no such effect. Effects on CYP1B1 expression and 4-MeOE(1/2) formation were less pronounced; only TCDD caused an induction, whereas PCB 169 was a potent and selective inhibitor of 4-MeOE(1/2) formation (IC(50) 0.7 and 2.2 nM PCB 169 in MCF-7 and MCF-10A cells, respectively). MCF-10A cells were less responsive toward dioxin-like compounds and the apparent EC(50) values for CYP1A1 and CYP1B1 induction in this study were 10-100 fold higher than in MCF-7 cells. The constitutive 4-/2-MeOE(1/2) ratios were 2.99 +/- 0.78 and 0.93 +/- 0.40 in MCF-7 and MCF-10A, respectively. Incubation with dioxin-like compounds resulted in a concentration-dependent decrease in the 4-/2-MeOE(1/2) ratio, but an increase in potentially carcinogenic estrogen metabolites in both MCF-7 and MCF-10A cells. This indicates that even though the 4-/2-OHE(1/2) ratio may be used as indicator for the presence of neoplasms, it is readily lowered by dioxin-like compounds and its value as a prognostic parameter for cancer risk should be further examined. PMID- 12902196 TI - Evaluation of nonthreshold leukemogenic response to methyl nitrosourea in p53 deficient C3H/He mice. AB - The classic controversy of whether genotoxic chemicals induce cancers with or without a certain low-dose limit, i.e., the threshold, is revisited because of a number of current publications available addressing the plausibility of "practical" thresholds even for genotoxic carcinogens, the mechanism of which may be hypothesized to be due, in part, to a repair system composed of ordinarily available various defense mechanisms under the steady-state DNA damage. The question of whether an absolute nonthreshold or a relative nonthreshold, i.e., a "practical" threshold specifically in the low-dose level, is present may not be answered even with the use of a prohibitively large number of wild-type mice. Could the excessive incidence of tumorigenesis in p53-deficient mice contribute to our understanding of the threshold vs nonthreshold issue in genotoxic carcinogenesis? This is considered because an exaggeration of tumorigenesis in p53-deficient mice is hypothesized to reduce or eliminate the range of threshold due to the p53-deficiency-mediated reduction of DNA repair and apoptosis. The present study of chemical leukemogenesis in p53-deficient mice by transplantation assay was designed to answer this question. Briefly, 218 C3H/He mice were lethally irradiated and repopulated with bone marrow cells from wild-type, heterozygous p53-deficient, and homozygous p53-deficient C3H/He mice. This was followed by treatment with a single and graded dose of methyl nitrosourea at 6.6, 14.8, 33.3, 50.0, and 75.0 mg/kg body wt, with the vehicle-treated control groups treated with zero dose for each genotype. Whereas mice repopulated with p53 deficient bone marrow cells showed a marked reduction of the threshold for leukemogenicity, mice repopulated with wild-type bone marrow cells did not exhibit leukemia at a dose of 33.3 mg/kg body wt and showed a curve with a high probability for the linear regression model with a positive dose intercept, predicting a threshold by the likelihood ratio test. Thus, the failure of wild type mice to show an increase in incidence of leukemogenesis at low doses of genotoxic carcinogens may be due not to a statistical rarity, but to various p53 related pharmacophysiological functions, possibly including DNA repair and apoptosis that may account for a threshold. PMID- 12902197 TI - 4-Methylthioamphetamine-induced hyperthermia in mice: influence of serotonergic and catecholaminergic pathways. AB - 4-Methylthioamphetamine (4-MTA), also known as p-methylthioamphetamine, is a new amphetamine derivative which in humans has been increasingly associated with severe intoxications and several deaths. As hyperthermia is considered to be one of the most life-threatening acute physiological consequences of amphetamine related intoxications, it was our aim to determine whether 4-MTA induces changes in body temperature in a mouse model. Accordingly, we measured the subcutaneous temperature after acute administration of 4-MTA in CD1 mice. Because hyperthermia seems to result from the central and peripheral actions of catecholamines and serotonin (5-hydroxytriptamine or 5-HT), we also investigated the possible interactions of some catecholaminergic and serotonergic receptor blockers and the inhibition of monoamine oxidase (MAO) with this effect. 4-MTA induced hyperthermia in CD1 mice. Blockade of the 5-HT receptors with methysergide and MAO inhibition with pargyline resulted in the potentiation of the 4-MTA-induced hyperthermic effect. Blockade of the alpha(1)-adrenergic receptors with prazosin completely reverted the 4-MTA-induced hyperthermia while with the beta-adrenergic receptor blocker dl-propranolol this reversal was not complete. Blockade of the alpha(2)-adrenergic receptors with yohimbine had no effect on the hyperthermia induced by 4-MTA. These results suggest that 4-MTA-induced hyperthermia is highly influenced by the catecholaminergic and serotonergic receptor activation and the MAO activity. PMID- 12902198 TI - Analysis of specific lysine histone H3 and H4 acetylation and methylation status in clones of cells with a gene silenced by nickel exposure. AB - We have previously reported that the gpt transgene in G12 Chinese hamster cells could be silenced by water-insoluble nickel compounds nickel sulfide (NiS) or nickel subsulfide (Ni(3)S(2)) and showed that the transgene was silenced by de novo DNA methylation and chromatin condensation. To further understand the nature of this silencing, we used the chromatin immunoprecipitation assay to elucidate the chromatin structure in nickel-induced silenced G12 clones. We also analyzed the effects of the DNA methyltransferase inhibitor 5-azacytidine (5-AzaC) and a histone deacetylase inhibitor trichostatin A (TSA) on histone H3 and H4 acetylation and gpt gene expression in selected nickel-silenced clones. We observed that both histone H3 and H4 were hypoacetylated and a methyl DNA-binding protein MeCP2 was targeted to the gpt gene locus, resulting in a localized inactive chromatin configuration in nickel-silenced cell clones. The histone H3K9 was also found methylated in three of four nickel- silenced cell clones, whereas the histone H3K9 was deacetylated in all four cell clones, indicating that the H3K9 methylation was involved in nickel-induced gene silencing. The acetylation of the gpt gene could be increased by a combination of 5-AzaC and TSA treatment, but not by either 5-AzaC or TSA alone. The gpt transcript was studied by either Northern blot or by semiquantitative RT-PCR following treatment of the silenced clones with TSA or 5-AzaC. An increase in gpt mRNA could be detected by RT-PCR in the clones that regained acetylation of H3 and H4. These data show that gene silencing induced by nickel in the gpt transgenic cell line involved a loss of histone acetylation and an activation of histone methylation. Both H4 and H3 histone acetylation were lost in the silenced clones and these clones exhibited an increase in the methylation of the lysine 9 in histone H3. PMID- 12902199 TI - Effect of polystyrene particles on lung microvascular permeability in isolated perfused rabbit lungs: role of size and surface properties. AB - The aim of this study was to investigate the role of particle number, total surface area, mass and surface chemical groups in (K(f,c)) changes. The lung effects of four different fine (110 nm) and ultrafine (24 nm) polystyrene particles have been tested in an isolated perfused rabbit lung model. Pulmonary microvascular permeability (K(f,c)) modifications were measured in response to intratracheal particle challenge. Polystyrene particles, mainly located in alveolar spaces and macrophages, induced a K(f,c) increase that was related to the total surface area and number of particles rather than to the instilled mass. Moreover, the positively charged amine-modified polystyrene particles were more effective in the K(f,c) response than the negatively charged carboxylate-modified polystyrene particles. We concluded that particle number and diameter that mathematically equally determined total surface area do not have the same importance in explaining the biological effects observed and that particle number could be an alternative to total surface area to describe the particle exposure. Furthermore, surface properties of polystyrene particles need to be considered to investigate the microvascular permeability changes measured in our model. PMID- 12902201 TI - Insulin-like growth factors (IGFs) and IGF-binding proteins in nephrotic syndrome children on glucocorticoid. AB - Growth failure is often seen in nephrotic children who fail to respond to glucocorticoid therapy or are glucocorticoid dependent. The underlying mechanism responsible for such growth defect remains poorly understood although long-term glucocorticoid use or nephrotic syndrome (NS) has been indicated to independently affect growth. Recent clinical evidence has suggested that changes in the serum levels of insulin-like growth factors (IGFs) and IGF-binding proteins (IGFBPs) among these children. The loss of insulin-like growth factor-I (IGF-1) and IGFBP 3 found in nephrotic children may directly prompt growth defect and retardation. In addition, glucocorticoid is believed to be associated with overt elevation of serum IGF-I levels suggesting potential development of IGF resistance, one of the main factors responsible for persistent growth retardation. Growth hormone therapy has been shown to effectively reconcile this IGF resistance. The present review will discuss the complex relationship among IGF, IGFBPs, and glucocorticoid in children with NS. PMID- 12902200 TI - Detection of viral infection in the respiratory tract of virus antibody free mice: advantages of high-resolution imaging for respiratory toxicology. AB - Using a highly sensitive membrane permeability assay, a viral infection was discovered in the lungs of virus antibody free (VAF) Swiss-Webster mice purchased for respiratory toxicology studies. The assay is based on the uptake of a charged fluorescent compound by cells lacking an intact plasma membrane. Lungs from 74% of the untreated animals from a single vendor tested positive for injury in this assay. High-resolution histopathologic analysis of 1-microm epoxy resin sections from affected animals identified increased peribronchiolar lymphocytic infiltration and markers of epithelial cell injury. Viral particles were directly observed to be budding from the membranes of infiltrating lymphocytic cells by transmission electron microscopy. Standard histological analysis of paraffin embedded tissues from lungs of the same mice failed to detect obvious pathology. Serological analyses failed to detect the presence of a virus in the affected mice. Therefore, we conclude that (1) a pathogenic condition was present in the respiratory systems of mice judged pathogen free by standard methodologies, (2) the observed condition produced a pattern of injury comparable to those caused by pulmonary toxicants, (3) high-resolution histopathology and advanced imaging techniques can increase the potential for detection of pathological conditions, and (4) apparently healthy animals can have unrecognized infections with the potential for confounding respiratory toxicology studies. PMID- 12902202 TI - Effects of the prilocaine and of the association of prilocaine with different vasoconstrictors on the isolated hearts of rats. AB - This study aimed to observe the effects of prilocaine chloridate (P) and the associations of P with felypressin (P+F), and epinephrine (P+E) on the heart rate and force of contraction on the isolated hearts of rats. The hearts were perfused according to Langendorff's method. The experiments were performed with the following substances: prilocaine (P), felypressin (F), and epinephrine (E). P was used in doses of 1.25, 2.5, and 5.0 mg. The same doses of F and P (1.0, 1.5 and 3.0 microg) were used in association with the P, respectively. Right after the application of the three doses of P, a reduction in the force of contraction was observed, with higher intensity at a higher dose, which led to cardiac arrest. A similar result was obtained with the administration of the three associations of P+F. Also the application of the three associations P+E resulted in a reduction of the force of contraction, that was, however, less intense with the associations of 1.25 mg P + 1.0 microg E and 2.5 mg P+1.5 microg E, as compared to the same doses of P alone and P+F. The two higher doses of P and P+F on the heart rate elicited a significant reduction in relation to the control, but with the associations of P+E no alterations were observed, with any one of the doses used. The results obtained in the present study suggest that an association of P and E must be used in local anesthesia, since epinephrine may well protect the heart from the depressive effects of the anesthetic salts. PMID- 12902203 TI - Age-related smooth muscle reactivity changes in the rat bladder: an in vitro study. AB - An experimental study was conducted to investigate developmental changes of the rat detrusor smooth muscle (DSM) reactivity from newborn to adult period. Urinary bladders were obtained from adult (4 months old), 1-month-old and newborn (4-7 days old) male Sprague-Dawley rats. DSM reactivity of the three groups was evaluated in organ chambers. The newborn DSM strips revealed markedly increased purinoceptor- and cholinoceptor-mediated contractions (ATP, carbachol) with increased maximum response (2.98- and 8.96-fold increase for ATP, 2.90- and 4.22 fold increase for carbachol, when the newborn bladder compared with 1-month-old and adult groups, respectively) and sensitivity (1.65- and 1.29-fold increase for ATP when the newborn bladder compared with the 1-month-old and adult groups, respectively). Additionally the maximum contractile response to KCl in the bladder from the newborn was 1.65- and 8.96-fold increased compared to bladders of the 1-month-old and adult groups, respectively. However, no significant changes in the adrenoceptor-mediated relaxation (isoproterenol) of the rat DSM were observed among the groups. These results indicate that development alters the in vitro responsiveness of rat DSM. The newborn rat bladder gains some of the adult bladder properties within 1 month. These changes are likely to reflect the changing role of parasympathetic regulation in the DSM reactivity during development of the rat bladder. PMID- 12902204 TI - The effects of paroxetine on rat isolated vas deferens. AB - The aim of the present study is to evaluate whether paroxetine (a selective serotonin re-uptake inhibitor) can modify the contractile responses of isolated vas deferens. Some contractile agents, potassium chloride (KCl), adenosine 5' triphosphate (ATP), noradrenaline (NA), and electrical field stimulation (EFS) caused contractions both in epididymal and prostatic portions of vas deferens. Paroxetine (PX) in concentrations 10(-7) and 10(-6)M potentiated the contractions to KCl and ATP only in epididymal portion but in higher concentrations (10(-5) and 10(-4)M) inhibited the responses in both portions. NA responses were inhibited by PX in all concentrations used, both in prostatic and epididymal portions. Prazosin (PR), an alpha adrenergic receptor blocking agent, inhibited PX-induced potentiation observed for higher concentrations of KCl. PR also inhibited PX-induced potentiation on the responses to ATP in epididymal portion. Pretreatment with PX (10(-7) to 10(-6)M) increased the contractions to EFS but in 10(-5) and 10(-4)M concentrations inhibited them. Even though the preparations were washed out, the inhibited responses of contractile agents could not be restored. After a washout period for PX, when Bay K 8644 (calcium channel activator) was added to the bath medium, the contractile responses to KCl were partially restored. In calcium-free medium, KCl caused contractions in concentrations higher than 80 mM with lower amplitudes which were not affected by PX. Reserpinization did not change the inhibitory pattern of PX's effect on exogenously applied NA in all concentrations tested. In reserpinized rats, the potentiation caused by PX in exogenously applied ATP responses was not observed. In conclusion, we can say that PX has two different effects: inhibition and potentiation of contractions to various agonists. The inhibitory effect of the drug can be explained by a calcium channel blocking activity. The potentiating effect of the drug is mainly related to its presynaptic action, such as NA re uptake inhibitory effect. PMID- 12902205 TI - P-glycoprotein inhibitors and their screening: a perspective from bioavailability enhancement. AB - Drug efflux pumps like P-glycoprotein (P-gp) and multidrug resistance (MDR) proteins were recognized to possess functional role in determining the pharmacokinetics of drugs administered by peroral as well as parenteral route. Advancements in molecular biology, to some extent, had revealed the structure, localization and functional role of P-glycoprotein and its mechanism of drug efflux. Broad substrate recognition by this protein and clinical implications of its inhibition has revolutionized cancer chemotherapy leading to design and development of novel P-glycoprotein inhibitors. In the recent times, the application of these inhibitors in improving peroral drug delivery has gained special interest. Inhibition of P-glycoprotein improves intestinal absorption and tissue distribution while reducing the substrate metabolism and its elimination. Eventually, various screening methodologies have been developed for determining the activity of P-glycoprotein, kinetics of drug transport and identification of substrates and inhibitors. In the present review, techniques used for screening P glycoprotein inhibitors and the scope of these inhibitors in optimizing peroral drug absorption and pharmacokinetics are discussed along with a brief introduction to P-glycoprotein, its physiological function and active role in extrusion of drugs. PMID- 12902206 TI - Evaluation of the antioxidant effect of melatonin by flow injection analysis luminol chemiluminescence. AB - The antioxidant effect of melatonin was evaluated by using a flow injection analysis chemiluminescence (FIA-CL) method to measure alone its direct antioxidant behaviour in aqueous solutions. Inhibition by melatonin, DL tryptophan or serotonin (5-hydroxytryptamine, 5-HT) of the steady CL emissions of various oxidants with luminol was measured. LogIC(50) values of melatonin were calculated: -2.6+/-0.1M for superoxide (O(2) radical(-)), -3.0+/-0.02 M, for hydrogen peroxide (H(2)O(2)), -4.7+/-0.04 M, for hypochlorous anion ((-)OCl), 2.8+/-0.1M, for hydoxyl (radical OH) and -2.0+/-0.03 M, for peroxynitrite (ONOO( )). Thus melatonin exerted a limited direct antioxidant activity towards reactive oxygen species (ROS) in aqueous solutions, broadly comparable to that of tryptophan, but less than that of serotonin and of major antioxidants such as ascorbic acid. This suggests that where the benefits of melatonin are correctly attributed to antioxidant behaviour, this is mediated largely through antioxidant enzymes. The pharmacological implications are discussed. PMID- 12902207 TI - Comparison of the cytotoxicity of two nitroheterocyclic drugs (NHCD) towards transformed and non-transformed cells. AB - The cytotoxicity of two nitroheterocyclic compounds (NHCD), Nitracrine, 1-nitro 9(3'3'-dimethylaminopropylamino) acridine (Polfa, Poland) and Quinifuryl, 2-(5' nitro-2'-furanyl) ethenyl-4-[N-[4-(N,N-diethylamino)-1'-methylbutyl] carbamoyl] quinoline (Dr. N. M. Sukhova, Institute of Organic Synthesis, Riga, Latvian Republic), towards two lines of leukaemic cells and a line of non-transformed cells, was determined under normoxia conditions. Although both drugs showed significant cytotoxicity to all cell lines (LC(50) for 24h, < or = 2 microM) with that of Nitracrine exceeding Quinifuryl, their toxicity towards murine leukaemia P388 was substantially higher, compared to murine fibroblasts NIH3T3. In addition, the rate of cell death was also two- to three-fold higher in case of P388 cells versus NIH3T3. Interestingly, human erythroleukaemia K562 cells were shown to uptake the drugs 10 min after their addition to the tissue culture medium, while the LC(50) values were reached after a substantial delay of 3h. This delay might be due to the intracellular transformation of drugs required for cell killing. PMID- 12902208 TI - Erdosteine prevents doxorubicin-induced cardiotoxicity in rats. AB - The clinical use of doxorubicin (Dxr) is limited by its cardiotoxic effects which are mediated by oxygen radicals. The purpose of this study was to investigate in vivo protective effects of erdosteine, an antioxidant agent because of its secondary active metabolites in vivo, against the cardiotoxicity induced by Dxr in rats. Three groups of male Sprague-Dawley rats (60 days old) were used. Group 1 was untreated group used as control; the other groups were treated with Dxr (single i.p. dosage of 20 mg kg(-1) b.wt.) or Dxr plus erdosteine (10 mg kg(-1) day(-1), orally), respectively. Erdosteine or oral saline treatment was done starting 2 days before Dxr for 12 days. The analyses were done at the 10th day of Dxr treatment. The protein carbonyl content, the activities of myeloperoxidase, aspartate aminotransferase (AST), lactate dehydrogenase (LDH), and creatine kinase (CK) as well as heart rate and blood pressures were significantly increased in Dxr group in comparison with the other groups. However, pulse pressure was decreased in Dxr group. The body and heart weights were decreased in both Dxr administered groups in comparison with control group. Disorganization of myocardial histology, picnotic nuclei, edema, and increase in collagen content around vessels were seen in the slides of Dxr group, whereas normal myocardial microscopy was preserved in Dxr plus erdosteine group. Collectively, these in vivo hemodynamic, enzymatic and morphologic studies provide an evidence for a possible prevention of cardiac toxicity in Dxr-treated patients. PMID- 12902209 TI - Evaluation of bioequivalence of isoniazid and pyrazinamide in three and four drugs fixed dose combinations using WHO simplified protocol. AB - The reliable supply of quality drugs in the form of fixed dose combination (FDC) is an essential part of tuberculosis treatment. The objective of this investigation was to evaluate whether the World Health Organization (WHO) simplified screening protocol for the bioequivalence assessment of rifampicin can be used for the evaluation of other components of FDC so as to ensure the bioavailability of all drugs at tissue site. These bioequivalence studies were conducted on 20 and 22 healthy male volunteers for evaluation of three and four drugs FDC formulations, respectively. Both studies were conducted as randomized, open, crossover trials and sampling schedule was upto 8h according to WHO recommended protocol for evaluation of rifampicin bioequivalence. The bioequivalence of isoniazid and pyrazinamide were estimated using AUC(0-8), AUC(0 alpha), and C(max). FDC formulation was considered bioequivalent to separate formulations for isoniazid and pyrazinamide if bioequivalence limit fall in between 0.80 and 1.25. Bioequivalence estimates of AUC(0-8) and AUC(0-alpha) for isoniazid and all the three pharmacokinetic measures of pyrazinamide were within the acceptable limits, whereas C(max) of isoniazid from four drugs FDC was outside the limit when evaluated by two-way ANOVA. After evaluation of isoniazid and pyrazinamide based on their pharmacokinetics, it was found that C(max) is being affected by limited sampling time points of WHO protocol. Further, AUC was a robust parameter unaffected by sampling schedule adopted. The WHO simplified protocol for assessment of rifampicin is also suitable for evaluating bioequivalence of isoniazid and pyrazinamide from FDC formulations. However, for comparison of rate of absorption by means of C(max), careful evaluation of concentration-time profile along with pharmacokinetics is necessary before final judgment. PMID- 12902210 TI - Influence of albendazole on the disposition kinetics and milk antimicrobial equivalent activity of enrofloxacin in lactating goats. AB - The pharmacokinetics of single intravenous and intramuscular administrations and milk antimicrobial equivalent activity of enrofloxacin at a dose of 5 mg per kilogram body weight were studied in clinically healthy lactating goats which were either not treated or had received 7.5 mg per kilogram body weight of albendazole orally. The concentrations of enrofloxacin in serum and milk were determined using microbiological assay. Following intravenous injection, enrofloxacin antimicrobial equivalent activity versus time data in serum was described by a two-compartmental open model. Albendazole treatment significantly decreased the elimination half-life (t(1/2beta)) (P>or=0.05) and the mean residence time (MRT) (P>or=0.05), whereas, the rate of enrofloxacin return to central compartment from peripheral tissue (K(21)) was significantly increased (P>or=0.01). In contrast, the volumes of distribution V(d(area)) and V(d(SS)) were significantly decreased (P>or=0.01 and P>or=0.05, respectively) in albendazole-treated goats. After intramuscular injection, enrofloxacin was rapidly absorbed in control and albendazole-treated lactating goats with absorption half-lives (t(1/2ab)) 0.43 and 0.39 h, respectively. The mean peaks of serum concentration (C(max)) were 0.68 and 0.65 mcg ml(-1) attained at (t(max)) 1.08 and 1.12 h, before and after albendazole dosing, respectively. The elimination half-life (t(1/2el)) and (MRT) following intramuscular injections were also shorter in the albendazole-treated lactating goats. The systemic bioavailability of enrofloxacin was significantly decreased from 110.16 to 84.38% in albendazole-treated lactating goats. Concomitant administration of albendazole with enrofloxacin resulted in significant alterations in the disposition kinetic of enrofloxacin and significant decrease in enrofloxacin concentrations in milk. Consequently, the interaction between albendazole and enrofloxacin could be of clinical significance and may require monitoring and adjustment of enrofloxacin dosage. PMID- 12902211 TI - The effects of caffeic acid phenethyl ester on tissue damage in lung after hindlimb ischemia-reperfusion. AB - AIM: The aim of this study was to investigate the effects of caffeic acid phenethyl ester (CAPE) on the lungs as a remote organ after performing hindlimb ischemia-reperfusion (I/R) and by assessing biochemical and histopathological analysis. METHODS: The animals were divided into three groups: control, I/R, and I/R with CAPE. I/R period for 8 h was performed on the right hindlimb of all the anesthesied rats in I/R and CAPE with I/R group. In the CAPE with I/R group, the animals received CAPE 10 microM by intraperitoneal injection 1h before the reperfusion. The animals in the control and I/R groups received a similar volume of saline solution by means of intraperitoneal injection. At the end of the reperfusion period, a midsternotomy was performed. Blood, bronchoalveolar lavage (BAL) and lung tissue were obtained, and were used for biochemical and histopathological examination. RESULTS: The tissue and serum malondyaldehyde levels were significantly lower in the control (P=0.0001 and 0.001, respectively) and in the CAPE with I/R groups (P=0.0001 and 0.003, respectively) compared to the I/R group. Tissue Na(+)-K(+) ATPase activity in the CAPE with I/R group was significantly higher than in the I/R group (P=0.0001). Reduced activity was found in the I/R group compared to the control group (P=0.0001). Myeloperoxidase activity (P=0.001) and protein concentration (P=0.034) in BAL were significantly reduced in CAPE-treated animals when compared with the I/R group. A decreased activity and protein concentration were found in the control group compared to the I/R group (P=0.0001 and 0.024, respectively). The lungs of the I/R group displayed intense peribronchial and perivascular leukocytic infiltration in histopathological examination compared to the CAPE with I/R group (P<0.05). CONCLUSION: CAPE seems to be effective in protecting remote organ injury caused by increased oxidative stress and neutrophil accumulation that results from an I/R injury. PMID- 12902212 TI - Risperidone-associated hyperprolactinemia: evaluation in twenty psychiatric outpatients. AB - Despite the lack of extrapyramidal side effects, some atypical antipsychotics can induce an increase in prolactinemia, as the conventional neuroleptics do. We decided to evaluate the effects of risperidone on serum prolactin levels and prolactin-related adverse effects in 20 outpatients of an Italian community psychiatric service. Patients enrolled in this study were on risperidone (2-8 mg per day; mean dose=4.15+/-0.4 mg per day) treatment in the period May-November 2002. The 20 patients, 13 women and 7 men (mean age=36.38+/-3.2 years for women and 29.7+/-2.2 for men) who accomplished inclusion criteria, participated in the study after giving informed written consent. Raised prolactin levels were observed in 13 (9 women and 4 men) out of 20 patients, but only 8 patients presented prolactin-related adverse effects, libido reduction being the most frequent. In this observational study, risperidone enhanced serum prolactin in 65% of patients. A good correlation was found between age and prolactin levels in pre-menopausal women, although no clear correlation among duration of treatment, dose used, prolactin levels and prolactin-related adverse effects could be established. PMID- 12902213 TI - Lack of a pharmacokinetic interaction between mirtazapine and the newer antipsychotics clozapine, risperidone and olanzapine in patients with chronic schizophrenia. AB - The effect of mirtazapine on steady-state plasma concentrations of the newer atypical antipsychotics clozapine, risperidone and olanzapine was investigated in 24 patients with chronic schizophrenia. In order to treat residual negative symptoms, additional mirtazapine (30 mg per day) was administered for six consecutive weeks to nine patients stabilized on clozapine therapy (200-650 mg per day), eight on risperidone (3-8 mg per day) and seven on olanzapine (10-20mg per day). There were only minimal and statistically insignificant changes in mean plasma concentrations of clozapine and its metabolite norclozapine, risperidone and its metabolite 9-hydroxyrisperidone, and olanzapine during the study period. Mirtazapine co-administration with either clozapine, risperidone or olanzapine was well tolerated. In the overall sample, a slight improvement in negative symptomatology, as assessed by the Scale for Assessment of Negative Symptoms, was observed at final evaluation (P<0.01) and six patients (two in each treatment group) were classified as responders. While double-blind, controlled studies are needed to evaluate the potential clinical benefits of mirtazapine in chronic schizophrenia, our findings indicate that mirtazapine has a negligible effect on the metabolism of clozapine, risperidone and olanzapine and can be added safely to an existing treatment with these antipsychotics. PMID- 12902215 TI - Characterization of the RpoS status of clinical isolates of Salmonella enterica. AB - The stationary-phase-inducible sigma factor, sigma(S) (RpoS), is the master regulator of the general stress response in Salmonella and is required for virulence in mice. rpoS mutants can frequently be isolated from highly passaged laboratory strains of Salmonella: We examined the rpoS status of 116 human clinical isolates of Salmonella, including 41 Salmonella enterica serotype Typhi strains isolated from blood, 38 S. enterica serotype Typhimurium strains isolated from blood, and 37 Salmonella serotype Typhimurium strains isolated from feces. We examined the abilities of these strains to produce the sigma(S) protein, to express RpoS-dependent catalase activity, and to resist to oxidative stress in the stationary phase of growth. We also carried out complementation experiments with a cloned wild-type rpoS gene. Our results showed that 15 of the 41 Salmonella serotype Typhi isolates were defective in RpoS. We sequenced the rpoS allele of 12 strains. This led to identification of small insertions, deletions, and point mutations resulting in premature stop codons or affecting regions 1 and 2 of sigma(S), showing that the rpoS mutations are not clonal. Thus, mutant rpoS alleles can be found in freshly isolated clinical strains of Salmonella serotype Typhi, and they may affect virulence properties. Interestingly however, no rpoS mutants were found among the 75 Salmonella serotype Typhimurium isolates. Strains that differed in catalase activity and resistance to hydrogen peroxide were found, but the differences were not linked to the rpoS status. This suggests that Salmonella serotype Typhimurium rpoS mutants are counterselected because rpoS plays a role in the pathogenesis of Salmonella serotype Typhimurium in humans or in the transmission cycle of the disease. PMID- 12902216 TI - Cloning of a nitrilase gene from the cyanobacterium Synechocystis sp. strain PCC6803 and heterologous expression and characterization of the encoded protein. AB - The gene encoding a putative nitrilase was identified in the genome sequence of the photosynthetic cyanobacterium Synechocystis sp. strain PCC6803. The gene was amplified by PCR and cloned into an expression vector. The encoded protein was heterologously expressed in the native form and as a His-tagged protein in Escherichia coli, and the recombinant strains were shown to convert benzonitrile to benzoate. The active enzyme was purified to homogeneity and shown by gel filtration to consist probably of 10 subunits. The purified nitrilase converted various aromatic and aliphatic nitriles. The highest enzyme activity was observed with fumarodinitrile, but also some rather hydrophobic aromatic (e.g., naphthalenecarbonitrile), heterocyclic (e.g., indole-3-acetonitrile), or long chain aliphatic (di-)nitriles (e.g., octanoic acid dinitrile) were converted with higher specific activities than benzonitrile. From aliphatic dinitriles with less than six carbon atoms only 1 mol of ammonia was released per mol of dinitrile, and thus presumably the corresponding cyanocarboxylic acids formed. The purified enzyme was active in the presence of a wide range of organic solvents and the turnover rates of dodecanoic acid nitrile and naphthalenecarbonitrile were increased in the presence of water-soluble and water-immiscible organic solvents. PMID- 12902214 TI - Sources of Campylobacter colonization in broiler chickens. PMID- 12902217 TI - Microbial and physiological characterization of weakly amylolytic but fast growing lactic acid bacteria: a functional role in supporting microbial diversity in pozol, a Mexican fermented maize beverage. AB - Pozol is an acid beverage obtained from the natural fermentation of nixtamal (heat- and alkali-treated maize) dough. The concentration of mono- and disaccharides from maize is reduced during nixtamalization, so that starch is the main carbohydrate available for lactic acid fermentation. In order to provide some basis to understand the role of amylolytic lactic acid bacteria (ALAB) in this fermented food, their diversity and physiological characteristics were determined. Forty amylolytic strains were characterized by phenotypic and molecular taxonomic methods. Four different biotypes were distinguished via ribotyping; Streptococcus bovis strains were found to be predominant. Streptococcus macedonicus, Lactococcus lactis, and Enterococcus sulfureus strains were also identified. S. bovis strain 25124 showed extremely low amylase yield relative to biomass (139 U g [cell dry weight](-1)) and specific rate of amylase production (130.7 U g [cell dry weight](-1) h(-1)). In contrast, it showed a high specific growth rate (0.94 h(-1)) and an efficient energy conversion yield to bacterial cell biomass (0.31 g of biomass g of substrate(-1)). These would confer on the strain a competitive advantage and are the possible reasons for its dominance. Transient accumulation of maltooligosaccharides during fermentation could presumably serve as energy sources for nonamylolytic species in pozol fermentation. This would explain the observed diversity and the dominance of nonamylolytic lactic acid bacteria at the end of fermentation. These results are the first step to understanding the importance of ALAB during pozol fermentation. PMID- 12902219 TI - Overexpression of the gene encoding GTP:mannose-1-phosphate guanyltransferase, mpg1, increases cellular GDP-mannose levels and protein mannosylation in Trichoderma reesei. AB - To elucidate the regulation and limiting factors in the glycosylation of secreted proteins, the mpg1 and dpm1 genes from Trichoderma reesei (Hypocrea jecorina) encoding GTP:alpha-D-mannose-1-phosphate guanyltransferase and dolichyl phosphate mannose synthase (DPMS), respectively, were overexpressed in T. reesei. No significant increases were observed in DPMS activity or protein secretion in dpm1 overexpressing transformants, whereas overexpression of mpg1 led to a twofold increase in GDP-mannose (GDPMan) levels. GDPMan was effectively utilized by mannnosyltransferases and resulted in hypermannosylation of secreted proteins in both N and O glycosylation. Overexpression of the mpg1 gene also increased the transcription of the dpm1 gene and DPMS activity. Our data indicate that the level of cellular GDPMan can play a major regulatory role in protein glycosylation in T. reesei. PMID- 12902218 TI - Comparing the dehalogenase gene pool in cultivated alpha-halocarboxylic acid degrading bacteria with the environmental metagene pool. AB - Culture-dependent and culture-independent approaches were used to determine the relationship between the dehalogenase gene pool in bacteria enriched and isolated on 2,2-dichloropropionic acid (22DCPA) and the environmental metagene pool (the collective gene pool of both the culturable and uncultured microbes) from which they were isolated. The dehalogenases in the pure-cultures isolates, which were able to degrade 22DCPA, were similar to previously described group I and II dehalogenases. Significantly, the majority of the dehalogenases isolated from activated sludge by degenerate PCR with primers specific for alpha-halocarboxylic acid dehalogenases were not closely related to the dehalogenases in any isolate. Furthermore, the dehalogenases found in the pure cultures predominated in the enrichments but were a minor component of the community used to inoculate the batch cultures. Phylogenetic analysis of the dehalogenase sequences isolated by degenerate PCR showed that the diversity of the group II deh gene was greater than that of the group I deh gene. Direct plating of the activated sludge onto minimal media supplemented with 22DCPA resulted in biomass and DNA from which dehalogenases were amplified. Analysis of the sequences revealed that they were much more closely related to the sequences found in the community used to start the enrichments. However, no pure cultures were obtained with this isolation method, and thus no pure cultures were available for identification. In this study we examined the link between genes found in pure cultures with the metagene pool from which they were isolated. The results show that there is a large bias introduced by culturing, not just in the bacteria isolated but also the degradative genes that they contain. Moreover, our findings serve as a caveat for studies involving the culturing of pure cultures of bacteria and conclusions which are drawn from analysis of these organisms. PMID- 12902220 TI - Vibrio harveyi nitroreductase is also a chromate reductase. AB - The chromate reductase purified from Pseudomonas ambigua was found to be homologous with several nitroreductases. Escherichia coli DH5alpha and Vibrio harveyi KCTC 2720 nitroreductases were chosen for the present study, and their chromate-reducing activities were determined. A fusion between glutathione S transferase (GST) and E. coli DH5alpha NfsA (GST-EcNfsA), a fusion between GST and E. coli DH5alpha NfsB (GST-EcNfsB), and a fusion between GST and V. harveyi KCTC 2720 NfsA (GST-VhNfsA) were prepared for their overproduction and easy purification. GST-EcNfsA, GST-EcNFsB, and GST-VhNFsA efficiently reduced nitrofurazone and 2,4,6-trinitrotoluene (TNT) as their nitro substrates. The K(m) values for GST-EcNfsA, GST-EcNfsB, and GST-VhNfsA for chromate reduction were 11.8, 23.5, and 5.4 micro M, respectively. The V(max) values for GST-EcNfsA, GST EcNfsB, and GST-VhNfsA were 3.8, 3.9, and 10.7 nmol/min/mg of protein, respectively. GST-VhNfsA was the most effective of the three chromate reductases, as determined by each V(max)/K(m) value. The optimal temperatures of GST-EcNfsA, GST-EcNfsB, and GST-VhNfsA for chromate reduction were 55, 30, and 30 degrees C, respectively. Thus, it is confirmed that nitroreductase can also act as a chromate reductase. Nitroreductases may be used in chromate remediation. GST EcNfsA, GST-EcNfsB, and GST-VhNfsA have a molecular mass of 50 kDa and exist as a monomer in solution. Thin-layer chromatography showed that GST-EcNfsA, GST EcNfsB, and GST-VhNfsA contain FMN as a cofactor. GST-VhNfsA reduced Cr(VI) to Cr(III). Cr(III) was much less toxic to E. coli than Cr(VI). PMID- 12902221 TI - Rhizobium leguminosarum biovar viciae 1-aminocyclopropane-1-carboxylate deaminase promotes nodulation of pea plants. AB - Ethylene inhibits nodulation in various legumes. In order to investigate strategies employed by Rhizobium to regulate nodulation, the 1-aminocyclopropane 1-carboxylate (ACC) deaminase gene was isolated and characterized from one of the ACC deaminase-producing rhizobia, Rhizobium leguminosarum bv. viciae 128C53K. ACC deaminase degrades ACC, the immediate precursor of ethylene in higher plants. Through the action of this enzyme, ACC deaminase-containing bacteria can reduce ethylene biosynthesis in plants. Insertion mutants with mutations in the rhizobial ACC deaminase gene (acdS) and its regulatory gene, a leucine-responsive regulatory protein-like gene (lrpL), were constructed and tested to determine their abilities to nodulate Pisum sativum L. cv. Sparkle (pea). Both mutants, neither of which synthesized ACC deaminase, showed decreased nodulation efficiency compared to that of the parental strain. Our results suggest that ACC deaminase in R. leguminosarum bv. viciae 128C53K enhances the nodulation of P. sativum L. cv. Sparkle, likely by modulating ethylene levels in the plant roots during the early stages of nodule development. ACC deaminase might be the second described strategy utilized by Rhizobium to promote nodulation by adjusting ethylene levels in legumes. PMID- 12902222 TI - Elucidation of the transmission patterns of an insect-borne bacterium. AB - Quantitative data on modes of transmission are a crucial element in understanding the ecology of microorganisms associated with animals. We investigated the transmission patterns of a gamma-proteobacterium informally known as pea aphid Bemisia-like symbiont (PABS), also known as T-type, which is widely but not universally distributed in natural populations of the pea aphid, Acyrthosiphon pisum. The vertical transmission of PABS to asexual and sexual morphs and sexually produced eggs was demonstrated by a diagnostic PCR-based assay, and the maximum estimated failure rate was 2%. Aphids naturally lacking PABS acquired PABS bacteria administered via the diet, and the infection persisted by vertical transmission for at least three aphid generations. PABS was also detected in two of five aphid honeydew samples tested and in all five siphuncular fluid samples tested but in none of 15 samples of salivary secretions from PABS-positive aphids. However, PABS-negative aphids did not acquire PABS when they were cocultured with PABS-positive aphids; the maximal estimated level of horizontal transmission was 18%. A deterministic model indicated that the force of infection by a horizontal transmission rate of 3% is sufficient to maintain a previously described estimate of the prevalence of PABS-positive aphids (37%), if the vertical transmission rate is 98%. We concluded that PABS infections in A. pisum can be maintained by high vertical transmission rates and occasional horizontal transmission, possibly via the oral route, in the absence of selection either for or against aphids bearing this bacterium. PMID- 12902223 TI - Application of the shsp gene, encoding a small heat shock protein, as a food grade selection marker for lactic acid bacteria. AB - Plasmid pSt04 of Streptococcus thermophilus contains a gene encoding a protein with homology to small heat shock proteins (A. Geis, H. A. M. El Demerdash, and K. J. Heller, Plasmid 50:53-69, 2003). Strains cured from the shsp plasmids showed significantly reduced heat and acid resistance and a lower maximal growth temperature. Transformation of the cloned shsp gene into S. thermophilus St11 lacking a plasmid encoding shsp resulted in increased resistance to incubation at 60 degrees C or pH 3.5 and in the ability to grow at 52 degrees C. A food-grade cloning system for S. thermophilus, based on the plasmid-encoded shsp gene as a selection marker, was developed. This approach allowed selection after transfer of native and recombinant shsp plasmids into different S. thermophilus and Lactococcus lactis strains. Using a recombinant plasmid carrying an erythromycin resistance (Em(r)) gene in addition to shsp, we demonstrated that both markers are equally efficient in selecting for plasmid-bearing cells. The average transformation rates in S. thermophilus (when we were selecting for heat resistance) were determined to be 2.4 x 10(4) and 1.0 x 10(4) CFU/0.5 micro g of DNA, with standard deviations of 0.54 x 10(4) and 0.32 x 10(4), for shsp and Em(r) selection, respectively. When we selected for pH resistance, the average transformation rates were determined to be 2.25 x 10(4) and 3.8 x 10(3) CFU/0.5 micro g of DNA, with standard deviations of 0.63 x 10(4) and 3.48 x 10(3), for shsp and Em(r) selection, respectively. The applicability of shsp as a selection marker was further demonstrated by constructing S. thermophilus plasmid pHRM1 carrying the shsp gene as a selection marker and the restriction-modification genes of another S. thermophilus plasmid as a functional trait. PMID- 12902224 TI - Multiplex real-time PCR for monitoring Heterobasidion annosum colonization in Norway spruce clones that differ in disease resistance. AB - A multiplex real-time PCR assay was developed to monitor the dynamics of the Picea abies-Heterobasidion annosum pathosystem. Tissue cultures and 32-year-old trees with low or high resistance to this pathogen were used as the host material. Probes and primers were based on a laccase gene for the pathogen and a polyubiquitin gene for the host. The real-time PCR procedure was compared to an ergosterol-based quantification method in a tissue culture experiment, and there was a strong correlation (product moment correlation coefficient, 0.908) between the data sets. The multiplex real-time PCR procedure had higher resolution and sensitivity during the early stages of colonization and also could be used to monitor the host. In the tissue culture experiment, host DNA was degraded more rapidly in the clone with low resistance than in the clone with high resistance. In the field experiment, the lesions elicited were not strictly proportional to the area colonized by the pathogen. Fungal colonization was more restricted and localized in the lesion in the clone with high resistance, whereas in the clone with low resistance, the fungus could be detected until the visible end of the lesion. Thus, the real-time PCR assay gives better resolution than does the traditionally used lesion length measurement when screening host clones for resistance. PMID- 12902225 TI - A new bacterial steroid degradation gene cluster in Comamonas testosteroni TA441 which consists of aromatic-compound degradation genes for seco-steroids and 3 ketosteroid dehydrogenase genes. AB - In Comamonas testosteroni TA441, testosterone is degraded via aromatization of the A ring, which is cleaved by the meta-cleavage enzyme TesB, and further degraded by TesD, the hydrolase for the product of TesB. TesEFG, encoded downstream of TesD, are probably hydratase, aldolase, and dehydrogenase for degradation of 2-oxohex-4-enoicacid, one of the products of TesD. Here we present a new and unique steroid degradation gene cluster in TA441, which consists of ORF18, ORF17, tesI, tesH, ORF11, ORF12, and tesDEFG. TesH and TesI are 3 ketosteroid-Delta(1)-dehydrogenase and 3-ketosteroid-Delta(4)(5alpha) dehydrogenase, respectively, which work in the early steps of steroid degradation. ORF17 probably encodes the reductase component of 9alpha-hydroxylase for 1,4-androstadiene-3,17-dione, which is the product of TesH in testosterone degradation. Gene disruption experiments showed that these genes are necessary for steroid degradation and do not have any isozymes in TA441. By Northern blot analysis, these genes were shown to be induced when TA441 was incubated with steroids (testosterone and cholic acid) but not with aromatic compounds [phenol, biphenyl, and 3-(3-hydroxyphenyl)propionic acid], indicating that these genes function exclusively in steroid degradation. PMID- 12902226 TI - Differences in susceptibility of Listeria monocytogenes strains to sakacin P, sakacin A, pediocin PA-1, and nisin. AB - Two hundred strains of Listeria monocytogenes collected from food and the food industry were analyzed for susceptibility to the class IIa bacteriocins sakacin P, sakacin A, and pediocin PA-1 and the class I bacteriocin nisin. The individual 50% inhibitory concentrations (IC(50)) were determined in a microtiter assay and expressed in nanograms per milliliter. The IC(50) of sakacin P ranged from 0.01 to 0.61 ng ml(-1). The corresponding values for pediocin PA-1, sakacin A, and nisin were 0.10 to 7.34, 0.16 to 44.2, and 2.2 to 781 ng ml(-1), respectively. The use of a large number of strains and the accuracy of the IC(50) determination revealed patterns not previously described, and for the first time it was shown that the IC(50) of sakacin P divided the L. monocytogenes strains into two distinct groups. Ten strains from each group were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of whole-cell proteins and amplified fragment length polymorphism. The results from these studies essentially confirmed the grouping based on the IC(50) of sakacin P. A high correlation was found between the IC(50) of sakacin P and that of pediocin PA-1 for the 200 strains. Surprisingly, the correlation between the IC(50) of the two class IIa bacteriocins sakacin A and sakacin P was lower than the correlation between the IC(50) of sakacin A and the class I bacteriocin nisin. PMID- 12902228 TI - Efficient library construction by in vivo recombination with a telomere originated autonomously replicating sequence of Hansenula polymorpha. AB - A high frequency of transformation and an equal gene dosage between transformants are generally required for activity-based selection of mutants from a library obtained by directed evolution. An efficient library construction method was developed by using in vivo recombination in Hansenula polymorpha. Various linear sets of vectors and insert fragments were transformed and analyzed to optimize the in vivo recombination system. A telomere-originated autonomously replicating sequence (ARS) of H. polymorpha, reported as a recombination hot spot, facilitates in vivo recombination between the linear transforming DNA and chromosomes. In vivo recombination of two linear DNA fragments containing the telomeric ARS drastically increases the transforming frequency, up to 10-fold, compared to the frequency of circular plasmids. Direct integration of the one-end recombined linear fragment into chromosomes produced transformants with single copy gene integration, resulting in the same expression level for the reporter protein between transformants. This newly developed in vivo recombination system of H. polymorpha provides a suitable library for activity-based selection of mutants after directed evolution. PMID- 12902227 TI - Purification and characterization of a novel mannitol dehydrogenase from a newly isolated strain of Candida magnoliae. AB - Mannitol biosynthesis in Candida magnoliae HH-01 (KCCM-10252), a yeast strain that is currently used for the industrial production of mannitol, is catalyzed by mannitol dehydrogenase (MDH) (EC 1.1.1.138). In this study, NAD(P)H-dependent MDH was purified to homogeneity from C. magnoliae HH-01 by ion-exchange chromatography, hydrophobic interaction chromatography, and affinity chromatography. The relative molecular masses of C. magnoliae MDH, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and size-exclusion chromatography, were 35 and 142 kDa, respectively, indicating that the enzyme is a tetramer. This enzyme catalyzed both fructose reduction and mannitol oxidation. The pH and temperature optima for fructose reduction and mannitol oxidation were 7.5 and 37 degrees C and 10.0 and 40 degrees C, respectively. C. magnoliae MDH showed high substrate specificity and high catalytic efficiency (k(cat) = 823 s( 1), K(m) = 28.0 mM, and k(cat)/K(m) = 29.4 mM(-1) s(-1)) for fructose, which may explain the high mannitol production observed in this strain. Initial velocity and product inhibition studies suggest that the reaction proceeds via a sequential ordered Bi Bi mechanism, and C. magnoliae MDH is specific for transferring the 4-pro-S hydrogen of NADPH, which is typical of a short-chain dehydrogenase reductase (SDR). The internal amino acid sequences of C. magnoliae MDH showed a significant homology with SDRs from various sources, indicating that the C. magnoliae MDH is an NAD(P)H-dependent tetrameric SDR. Although MDHs have been purified and characterized from several other sources, C. magnoliae MDH is distinguished from other MDHs by its high substrate specificity and catalytic efficiency for fructose only, which makes C. magnoliae MDH the ideal choice for industrial applications, including enzymatic synthesis of mannitol and salt tolerant plants. PMID- 12902229 TI - Spread of recombinant DNA by roots and pollen of transgenic potato plants, identified by highly specific biomonitoring using natural transformation of an Acinetobacter sp. AB - Transgenic potato plants with the nptII gene coding for neomycin phosphotransferase (kanamycin resistance) as a selection marker were examined for the spread of recombinant DNA into the environment. We used the recombinant fusion of nptII with the tg4 terminator for a novel biomonitoring technique. This depended on natural transformation of Acinetobacter sp. strain BD413 cells having in their genomes a terminally truncated nptII gene (nptII'; kanamycin sensitivity) followed by the tg4 terminator. Integration of the recombinant fusion DNA by homologous recombination in nptII' and tg4 restored nptII, leading to kanamycin-resistant transformants. DNA of the transgenic potato was detectable with high sensitivity, while no transformants were obtained with the DNA of other transgenic plants harboring nptII in different genetic contexts. The recombinant DNA was frequently found in rhizosphere extracts of transgenic potato plants from field plots. In a series of field plot and greenhouse experiments we identified two sources of this DNA: spread by roots during plant growth and by pollen during flowering. Both sources also contributed to the spread of the transgene into the rhizospheres of nontransgenic plants in the vicinity. The longest persistence of transforming DNA in field soil was observed with soil from a potato field in 1997 sampled in the following year in April and then stored moist at 4 degrees C in the dark for 4 years prior to extract preparation and transformation. In this study natural transformation is used as a reliable laboratory technique to detect recombinant DNA but is not used for monitoring horizontal gene transfer in the environment. PMID- 12902230 TI - Diversity and seasonal variability of beta-Proteobacteria in biofilms of polluted rivers: analysis by temperature gradient gel electrophoresis and cloning. AB - The beta-subgroup of the Proteobacteria has been shown to be important in aquatic habitats and was investigated in depth here by molecular 16S rRNA techniques in biofilms of the Elbe River and its polluted tributary, the Spittelwasser River. The bacterial 16S rRNA genes were cloned from each site, screened for beta proteobacterial clones and sequenced. River biofilm clones from both rivers grouped into 9 clusters (RBFs). RBFs 1, 2, and 3 fell into the recently described betaI cluster of cosmopolitan freshwater bacteria, where they represented new species related to Rhodoferax, Aquaspirillum, and Hydrogenophaga: RBFs 4 to 7 affiliated with Aquabacterium commune, Ideonella dechloratans, and Sphaerotilus natans, respectively. The two remaining RBFs were uncultivated clusters, one of them being distantly related to Gallionella ferruginea. Seasonal changes in the relative intensity of the beta-proteobacterial 16S rRNA genes of biofilms harvested monthly for 1 year were determined by specific amplification and separation by temperature gradient gel electrophoresis (TGGE). Bands were identified by comparison of clones to community fingerprints by TGGE. Eight of 13 identified bands were shared by both habitats but showed different relative abundance and seasonal variability in the two rivers, probably caused by differences in temperature and pollutants. The data indicate new not-yet cultivated clusters of river biofilm organisms, some of them probably distributed globally. They confirm the importance of certain known freshwater genera in river biofilms. The high phylogenetic resolution obtained by clone library analysis combined with the high temporal resolution obtained by TGGE suggest that the observed microdiversity in the river biofilm clone libraries might be caused by phylogenetically closely related microbial populations which are adapted to ecological parameters. PMID- 12902231 TI - Role of capsular colanic acid in adhesion of uropathogenic Escherichia coli. AB - Urinary tract infections are the most common urologic disease in the United States and one of the most common bacterial infections of any organ system. Biofilms persist in the urinary tract and on catheter surfaces because biofilm microorganisms are resistant to host defense mechanisms and antibiotic therapy. The first step in the establishment of biofilm infections is bacterial adhesion; preventing bacterial adhesion represents a promising method of controlling biofilms. Evidence suggests that capsular polysaccharides play a role in adhesion and pathogenicity. This study focuses on the role of physiochemical and specific binding interactions during adhesion of colanic acid exopolysaccharide mutant strains. Bacterial adhesion was evaluated for isogenic uropathogenic Escherichia coli strains that differed in colanic acid expression. The atomic force microscope (AFM) was used to directly measure the reversible physiochemical and specific binding interactions between bacterial strains and various substrates as bacteria initially approach the interface. AFM results indicate that electrostatic interactions were not solely responsible for the repulsive forces between the colanic acid mutant strains and hydrophilic substrates. Moreover, hydrophobic interactions were not found to play a significant role in adhesion of the colanic acid mutant strains. Adhesion was also evaluated by parallel-plate flow cell studies in comparison to AFM force measurements to demonstrate that prolonged incubation times alter bacterial adhesion. Results from this study demonstrate that the capsular polysaccharide colanic acid does not enhance bacterial adhesion but rather blocks the establishment of specific binding as well as time-dependent interactions between uropathogenic E. coli and inert substrates. PMID- 12902232 TI - Production and properties of bacteriocin-like inhibitory substances from the swine pathogen Streptococcus suis serotype 2. AB - Streptococcus suis serotype 2 is a major pathogen found in the upper respiratory tract of swine. In this study, isolates of this bacterial species were tested for the production of bacteriocin-like inhibitory substances (BLIS). Of the 38 strains tested, four inhibited the growth of other S. suis isolates according to a deferred-antagonism plate assay. Interestingly, three of the strains were originally isolated from healthy carrier pigs and were considered nonvirulent. Three isolates (94-623, 90-1330, and AAH4) that produced BLIS in liquid broth were selected for further characterization. None of the inhibitory activities was related to the production of either organic acids or hydrogen peroxide. The BLIS produced by these strains were heat stable and proteinase K, pronase, and elastase sensitive but were trypsin and chymotrypsin resistant. They were stable at pH 2 and 12 and had molecular masses in the range of 14 to 30 kDa. Maximum production was observed during the mid-log phase. Following a curing procedure with novobiocin, only 90-1330 lost the ability to produce BLIS, suggesting that the BLIS might be plasmid encoded. Analysis of the inhibitory spectra revealed that the BLIS-producing strains also inhibited the growth of Actinobacillus minor, Actinobacillus porcinus, Enterococcus durans, Micrococcus luteus, Streptococcus agalactiae, Streptococcus dysgalactiae subsp. dysgalactiae, Streptococcus equi subsp. zooepidemicus, and S. dysgalactiae subsp. equisimilis. This study reports for the first time the ability of the swine pathogen S. suis serotype 2 to produce BLIS with the characteristics of classic bacteriocins. Further studies are required to investigate the possibility of using bacteriocin producing strains to prevent swine infections caused by virulent strains of S. suis serotype 2. PMID- 12902233 TI - Preslaughter holding environment in pork plants is highly contaminated with Salmonella enterica. AB - The objective of this study was to determine whether abattoir pens can provide a Salmonella enterica infection source during the 2 to 4 h of preharvest holding. Previous work has suggested that pigs may be getting infected, but little has been reported on the environmental contamination of abattoir holding pens. For 24 groups of pigs studied ( approximately 150 animals/group) at two high-capacity abattoirs, six pooled fecal samples (n, 10 per pool) were collected from each transport trailer immediately after pigs were unloaded. Holding pens were sampled (one drinking water sample and six pooled floor samples consisting of swabs, residual liquid, and feces) prior to entry of study pigs for the routine holding period ( approximately 2.5 h). After slaughter, cecal contents and ileocecal lymph nodes were collected, on the processing line, from 30 pigs in each studied group. All samples were cultured for the isolation and identification of S. enterica by primary enrichment in GN-Hajna and tetrathionate broths, secondary enrichment in Rappaport-Vassiliadis broth, and plating on brilliant green sulfa and xylose-lysine-tergitol-4 agars, followed by biochemical and serological identification. The study pens were highly contaminated with S. enterica; all holding pens sampled had at least one positive sample. Additionally, 33% (8 of 24) of drinking water samples were positive for S. enterica. All 24 groups of pigs had S. enterica-positive cecal contents and ileocecal lymph nodes, including those groups from transport trailers with no positive samples. From pigs, trailers, and pens, 586 isolates representing 36 different Salmonella serovars were isolated. Of the 353 isolates from pigs (109 from ileocecal lymph nodes plus 244 from cecal contents), 19% were identified as belonging to the same serovars as those isolated from the respective pens; 27% were identified as belonging to the same serovars as those isolated from the trailers. Sixteen percent of the unique serovars were isolated from both pigs and pens, suggesting that pens served as the infection source. This study demonstrates highly contaminated abattoir holding pens and watering sources. It also demonstrates that holding pens can serve as an infection source. This study identifies the abattoir holding pens as a significant hazard and a potential control point for Salmonella contamination in the preharvest pork production chain. PMID- 12902234 TI - Molecular characterization of microsporidia indicates that wild mammals Harbor host-adapted Enterocytozoon spp. as well as human-pathogenic Enterocytozoon bieneusi. AB - Over 13 months, 465 beavers, foxes, muskrats, otters, and raccoons were trapped in four counties in eastern Maryland and examined by molecular methods for microsporidia. A two-step nested PCR protocol was developed to amplify a 392-bp fragment of the internal transcribed spacer region of the rRNA gene of Enterocytozoon spp., with the use of primers complementary to the conserved regions of published nucleotide sequences. Fifty-nine PCR-positive samples were sequenced. Multiple alignments of these sequences identified 17 genotypes of Enterocytozoon spp. (WL1 to WL17); of these, 15 have not been reported before. Most of the genotypes were found in multiple species of wildlife and belonged to a major group consisting of all the previously described Enterocytozoon bieneusi genotypes from human and domestic animals. Some of the isolates from muskrats and raccoons formed two distinct groups. Results of this study indicate that fur bearing mammals, especially those closely associated with surface water, can be a potential source of human-pathogenic E. bieneusi. However, there are also host adapted Enterocytozoon genotypes in wildlife, which may represent species different from E. bieneusi and have no apparent public health significance. This is the first report of E. bieneusi in wildlife. PMID- 12902235 TI - PCR assay of the groEL gene for detection and differentiation of Bacillus cereus group cells. AB - Strains of species in the Bacillus cereus group are potentially enterotoxic. Thus, the detection of all B. cereus group strains is important. As 16S ribosomal DNA sequence analysis cannot adequately differentiate species of the B. cereus group, we explored the potential of the groEL gene as a phylogenetic marker. A phylogenetic analysis of the groEL sequences of 78 B. cereus group strains revealed that the B. cereus group strains were split into two major clusters, one including six B. mycoides and one B. pseudomycoides (cluster II) and the other including two B. mycoides and the rest of the B. cereus group strains (cluster I). Cluster I was further differentiated into two subclusters, Ia and Ib. The sodA gene sequences of representative strains from different clusters were also compared. The phylogenetic tree constructed from the sodA sequences showed substantial similarity to the tree constructed from the groEL sequences. Based on the groEL sequences, a PCR assay for detection and identification of B. cereus group strains was developed. Subsequent restriction fragment length polymorphism (RFLP) analysis verified the PCR amplicons and the differentiation of the B. cereus group strains. RFLP with MboI was identical for all the B. cereus group strains analyzed, while RFLP with MfeI or PstI classified all B. cereus and B. thuringiensis strains into two groups. All cluster II B. mycoides and B. pseudomycoides strains could be discriminated from other B. cereus group bacteria by restriction analysis with TspRI. PMID- 12902236 TI - Isolation and characterization of Campylobacter bacteriophages from retail poultry. AB - The ability of phages to survive processing is an important aspect of their potential use in the biocontrol of Campylobacter in poultry production. To this end, we have developed a procedure to recover Campylobacter bacteriophages from chilled and frozen retail poultry and have validated the sensitivity of the method by using a characterized Campylobacter phage (i.e., NCTC 12674). By using this method, we have shown that Campylobacter phages can survive on retail chicken under commercial storage conditions. Retail chicken portions purchased in the United Kingdom were screened for the presence of endogenous Campylobacter phages. Thirty-four Campylobacter bacteriophages were isolated from 300 chilled retail chicken portions, but none could be recovered from 150 frozen chicken portions. The phage isolates were characterized according to their lytic profiles, morphology, and genome size. The free-range products were significantly more likely to harbor phages (P < 0.001 by single-factor analysis of variance) than were standard or economy products. This study demonstrates that Campylobacter bacteriophages, along with their hosts, can survive commercial poultry processing procedures and that the phages exhibited a wide range of recovery rates from chicken skin stored at 4 degrees C. PMID- 12902237 TI - Biocontrol of Listeria monocytogenes on fresh-cut produce by treatment with lytic bacteriophages and a bacteriocin. AB - The fresh-cut produce industry has been the fastest-growing portion of the food retail market during the past 10 years, providing consumers with convenient and nutritious food. However, fresh-cut fruits and vegetables raise food safety concerns, because exposed tissue may be colonized more easily by pathogenic bacteria than intact produce. This is due to the higher availability of nutrients on cut surfaces and the greater potential for contamination because of the increased amount of handling. We found that applied Listeria monocytogenes populations survived and increased only slightly on fresh-cut Red Delicious apples stored at 10 degrees C but increased significantly on fresh-cut honeydew melons stored at 10 degrees C over 7 days. In addition, we examined the effect of lytic, L. monocytogenes-specific phages via two phage application methods, spraying and pipetting, on L. monocytogenes populations in artificially contaminated fresh-cut melons and apples. The phage mixture reduced L. monocytogenes populations by 2.0 to 4.6 log units over the control on honeydew melons. On apples, the reduction was below 0.4 log units. In combination with nisin (a bacteriocin), the phage mixture reduced L. monocytogenes populations by up to 5.7 log units on honeydew melon slices and by up to 2.3 log units on apple slices compared to the control. Nisin alone reduced L. monocytogenes populations by up to 3.2 log units on honeydew melon slices and by up to 2.0 log units on apple slices compared to the control. The phage titer was stable on melon slices, but declined rapidly on apple slices. The spray application of the phage and phage plus nisin reduced the bacterial numbers at least as much as the pipette application. The effectiveness of the phage treatment also depended on the initial concentration of L. monocytogenes. PMID- 12902238 TI - Osmoregulation in the parasitic protozoan Tritrichomonas foetus. AB - Tritrichomonas foetus was shown to undergo a regulatory volume increase (RVI) when it was subjected to hyperosmotic challenge, but there was no regulatory volume decrease after hypoosmotic challenge, as determined by using both light scattering methods and measurement of intracellular water space to monitor cell volume. An investigation of T. foetus intracellular amino acids revealed a pool size (65 mM) that was similar to that of Trichomonas vaginalis but was considerably smaller than those of Giardia intestinalis and Crithidia luciliae. Changes in amino acid concentrations in response to hyperosmotic challenge were found to account for only 18% of the T. foetus RVI. The T. foetus intracellular sodium and potassium concentrations were determined to be 35 and 119 mM, respectively. The intracellular K(+) concentration was found to increase considerably during exposure to hyperosmotic stress, and, assuming that there was a monovalent accompanying anion, this increase was estimated to account for 87% of the RVI. By using light scattering it was determined that the T. foetus RVI was enhanced by elevated external K(+) concentrations and was inhibited when K(+) and/or Cl(-) was absent from the medium. The results suggested that the well documented Na(+)-K(+)-2Cl(-) cotransport system was responsible for the K(+) influx activated during the RVI. However, inhibitors of Na(+)-K(+)-2Cl(-) cotransport in other systems, such as quinine, ouabain, furosemide, and bumetanide, had no effect on the RVI or K(+) influx in T. foetus. PMID- 12902239 TI - Identification and characterization of phenylpyruvate decarboxylase genes in Saccharomyces cerevisiae. AB - Catabolism of amino acids via the Ehrlich pathway involves transamination to the corresponding alpha-keto acids, followed by decarboxylation to an aldehyde and then reduction to an alcohol. Alternatively, the aldehyde may be oxidized to an acid. This pathway is functional in Saccharomyces cerevisiae, since during growth in glucose-limited chemostat cultures with phenylalanine as the sole nitrogen source, phenylethanol and phenylacetate were produced in quantities that accounted for all of the phenylalanine consumed. Our objective was to identify the structural gene(s) required for the decarboxylation of phenylpyruvate to phenylacetaldehyde, the first specific step in the Ehrlich pathway. S. cerevisiae possesses five candidate genes with sequence similarity to genes encoding thiamine diphosphate-dependent decarboxylases that could encode this activity: YDR380w/ARO10, YDL080C/THI3, PDC1, PDC5, and PDC6. Phenylpyruvate decarboxylase activity was present in cultures grown with phenylalanine as the sole nitrogen source but was absent from ammonia-grown cultures. Furthermore, the transcript level of one candidate gene (ARO10) increased 30-fold when phenylalanine replaced ammonia as the sole nitrogen source. Analyses of phenylalanine catabolite production and phenylpyruvate decarboxylase enzyme assays indicated that ARO10 was sufficient to encode phenylpyruvate decarboxylase activity in the absence of the four other candidate genes. There was also an alternative activity with a higher capacity but lower affinity for phenylpyruvate. The candidate gene THI3 did not itself encode an active phenylpyruvate decarboxylase but was required along with one or more pyruvate decarboxylase genes (PDC1, PDC5, and PDC6) for the alternative activity. The K(m) and V(max) values of the two activities differed, showing that Aro10p is the physiologically relevant phenylpyruvate decarboxylase in wild-type cells. Modifications to this gene could therefore be important for metabolic engineering of the Ehrlich pathway. PMID- 12902241 TI - Ubiquity and persistence of Escherichia coli in a Midwestern coastal stream. AB - Dunes Creek, a small Lake Michigan coastal stream that drains sandy aquifers and wetlands of Indiana Dunes, has chronically elevated Escherichia coli levels along the bathing beach near its outfall. This study sought to understand the sources of E. coli in Dunes Creek's central branch. A systematic survey of random and fixed sampling points of water and sediment was conducted over 3 years. E. coli concentrations in Dunes Creek and beach water were significantly correlated. Weekly monitoring at 14 stations during 1999 and 2000 indicated chronic loading of E. coli throughout the stream. Significant correlations between E. coli numbers in stream water and stream sediment, submerged sediment and margin, and margin and 1 m from shore were found. Median E. coli counts were highest in stream sediments, followed by bank sediments, sediments along spring margins, stream water, and isolated pools; in forest soils, E. coli counts were more variable and relatively lower. Sediment moisture was significantly correlated with E. coli counts. Direct fecal input inadequately explains the widespread and consistent occurrence of E. coli in the Dunes Creek watershed; long-term survival or multiplication or both seem likely. The authors conclude that (i) E. coli is ubiquitous and persistent throughout the Dunes Creek basin, (ii) E. coli occurrence and distribution in riparian sediments help account for the continuous loading of the bacteria in Dunes Creek, and (iii) ditching of the stream, increased drainage, and subsequent loss of wetlands may account for the chronically high E. coli levels observed. PMID- 12902240 TI - Effects of cultivation conditions on folate production by lactic acid bacteria. AB - A variety of lactic acid bacteria were screened for their ability to produce folate intracellularly and/or extracellularly. Lactococcus lactis, Streptococcus thermophilus, and Leuconostoc spp. all produced folate, while most Lactobacillus spp., with the exception of Lactobacillus plantarum, were not able to produce folate. Folate production was further investigated in L. lactis as a model organism for metabolic engineering and in S. thermophilus for direct translation to (dairy) applications. For both these two lactic acid bacteria, an inverse relationship was observed between growth rate and folate production. When cultures were grown at inhibitory concentrations of antibiotics or salt or when the bacteria were subjected to low growth rates in chemostat cultures, folate levels in the cultures were increased relative to cell mass and (lactic) acid production. S. thermophilus excreted more folate than L. lactis, presumably as a result of differences in the number of glutamyl residues of the folate produced. In S. thermophilus 5,10-methenyl and 5-formyl tetrahydrofolate were detected as the major folate derivatives, both containing three glutamyl residues, while in L. lactis 5,10-methenyl and 10-formyl tetrahydrofolate were found, both with either four, five, or six glutamyl residues. Excretion of folate was stimulated at lower pH in S. thermophilus, but pH had no effect on folate excretion by L. lactis. Finally, several environmental parameters that influence folate production in these lactic acid bacteria were observed; high external pH increased folate production and the addition of p-aminobenzoic acid stimulated folate production, while high tyrosine concentrations led to decreased folate biosynthesis. PMID- 12902242 TI - Survival of Salmonella enterica in freshwater and sediments and transmission by the aquatic midge Chironomus tentans (Chironomidae: Diptera). AB - Survival of a nalidixic acid-resistant strain of Salmonella enterica serovar Typhimurium mr-DT-104 in water and sediments was tested using artificially contaminated aquaria. Water samples remained culture positive for salmonella for up to 54 days. Sediment samples were culture positive up to 119 days. In addition, potential mechanisms for spreading salmonella in the environments by chironomid larvae and adults were tested. We evaluated the acquisition of mr-DT 104 by chironomids from contaminated aquatic sediments and subsequent spread to uncontaminated sediments. Larval chironomids raised in contaminated sediments became culture positive, and the bacteria were carried over to adults after emergence. Contamination of clean sediments by chironomid larvae was not demonstrated. These findings clearly suggest that mr-DT-104 serovar organisms can survive in aquatic sediments for at least several months. Uptake of salmonellae by chironomid larvae and adults suggests that they are possible vectors of mr-DT 104 in both aquatic and terrestrial environments, although the role of larval defecation in movement of bacteria to new sediments was not demonstrated. PMID- 12902243 TI - Real-time PCR for simultaneous detection and quantification of Borrelia burgdorferi in field-collected Ixodes scapularis ticks from the Northeastern United States. AB - The density of spirochetes in field-collected or experimentally infected ticks is estimated mainly by assays based on microscopy. In this study, a real-time quantitative PCR (qPCR) protocol targeting the Borrelia burgdorferi-specific recA gene was adapted for use with a Lightcycler for rapid detection and quantification of the Lyme disease spirochete, B. burgdorferi, in field-collected Ixodes scapularis ticks. The sensitivity of qPCR for detection of B. burgdorferi DNA in infected ticks was comparable to that of a well-established nested PCR targeting the 16S-23S rRNA spacer. Of the 498 I. scapularis ticks collected from four northeastern states (Rhode Island, Connecticut, New York, and New Jersey), 91 of 438 (20.7%) nymphal ticks and 15 of 60 (25.0%) adult ticks were positive by qPCR assay. The number of spirochetes in individual ticks varied from 25 to 197,200 with a mean of 1,964 spirochetes per nymphal tick and a mean of 5,351 spirochetes per adult tick. No significant differences were found in the mean numbers of spirochetes counted either in nymphal ticks collected at different locations in these four states (P = 0.23 by one-way analysis of variance test) or in ticks infected with the three distinct ribosomal spacer restriction fragment length polymorphism types of B. burgdorferi (P = 0.39). A high degree of spirochete aggregation among infected ticks (variance-to-mean ratio of 24,877; moment estimate of k = 0.279) was observed. From the frequency distribution data and previously published transmission studies, we estimated that a minimum of 300 organisms may be required in a host-seeking nymphal tick to be able to transmit infection to mice while feeding on mice. These data indicate that real-time qPCR is a reliable approach for simultaneous detection and quantification of B. burgdorferi infection in field-collected ticks and can be used for ecological and epidemiological surveillance of Lyme disease spirochetes. PMID- 12902244 TI - Rapid identification of Candida species by using nuclear magnetic resonance spectroscopy and a statistical classification strategy. AB - Nuclear magnetic resonance (NMR) spectra were acquired from suspensions of clinically important yeast species of the genus Candida to characterize the relationship between metabolite profiles and species identification. Major metabolites were identified by using two-dimensional correlation NMR spectroscopy. One-dimensional proton NMR spectra were analyzed by using a staged statistical classification strategy. Analysis of NMR spectra from 442 isolates of Candida albicans, C. glabrata, C. krusei, C. parapsilosis, and C. tropicalis resulted in rapid, accurate identification when compared with conventional and DNA-based identification. Spectral regions used for the classification of the five yeast species revealed species-specific differences in relative amounts of lipids, trehalose, polyols, and other metabolites. Isolates of C. parapsilosis and C. glabrata with unusual PCR fingerprinting patterns also generated atypical NMR spectra, suggesting the possibility of intraspecies discontinuity. We conclude that NMR spectroscopy combined with a statistical classification strategy is a rapid, nondestructive, and potentially valuable method for identification and chemotaxonomic characterization that may be broadly applicable to fungi and other microorganisms. PMID- 12902245 TI - Engineering Deinococcus geothermalis for bioremediation of high-temperature radioactive waste environments. AB - Deinococcus geothermalis is an extremely radiation-resistant thermophilic bacterium closely related to the mesophile Deinococcus radiodurans, which is being engineered for in situ bioremediation of radioactive wastes. We report that D. geothermalis is transformable with plasmids designed for D. radiodurans and have generated a Hg(II)-resistant D. geothermalis strain capable of reducing Hg(II) at elevated temperatures and in the presence of 50 Gy/h. Additionally, D. geothermalis is capable of reducing Fe(III)-nitrilotriacetic acid, U(VI), and Cr(VI). These characteristics support the prospective development of this thermophilic radiophile for bioremediation of radioactive mixed waste environments with temperatures as high as 55 degrees C. PMID- 12902246 TI - Microbial quality and direct PCR identification of lactic acid bacteria and nonpathogenic Staphylococci from artisanal low-acid sausages. AB - Detection of six species of lactic acid bacteria and six species of gram-positive catalase-positive cocci from low-acid fermented sausages (fuets and chorizos) was assessed by species-specific PCR. Without enrichment, Lactobacillus sakei and Lactobacillus curvatus were detected in 11.8% of the samples, and Lactobacillus plantarum and Staphylococcus xylosus were detected in 17.6%. Enriched samples allowed the detection of L. sakei and S. xylosus in all of the samples (100%) and of Enterococcus faecium in 11.8% of the sausages. The percentages of L. curvatus, L. plantarum, Staphylococcus carnosus, and Staphylococcus epidermidis varied depending on the sausage type. L. curvatus was detected in 80% of fuets and in 57% of chorizos. L. plantarum was found in 50% of fuets and 100% of chorizos. S. epidermidis was detected in only 11.8% of fuets, and S. carnosus was detected in only 5.9% of chorizos. Lactococcus lactis, Staphylococcus warneri, and Staphylococcus simulans were not detected in any sausage type. From a microbiological point of view, 70.6% of the samples could be considered of high quality, as they had low counts of Enterobacteriaceae and did not contain any of the food-borne pathogens assayed. PMID- 12902247 TI - A newly discovered Bacteroides conjugative transposon, CTnGERM1, contains genes also found in gram-positive bacteria. AB - Results of a recent study of antibiotic resistance genes in human colonic Bacteroides strains suggested that gene transfer events between members of this genus are fairly common. The identification of Bacteroides isolates that carried an erythromycin resistance gene, ermG, whose DNA sequence was 99% identical to that of an ermG gene found previously only in gram-positive bacteria raised the further possibility that conjugal elements were moving into Bacteroides species from other genera. Six of seven ermG-containing Bacteroides strains tested were able to transfer ermG by conjugation. One of these strains was chosen for further investigation. Results of pulsed-field gel electrophoresis experiments showed that the conjugal element carrying ermG in this strain is an integrated element about 75 kb in size. Thus, the element appears to be a conjugative transposon (CTn) and was designated CTnGERM1. CTnGERM1 proved to be unrelated to the predominant type of CTn found in Bacteroides isolates-CTns of the CTnERL/CTnDOT family-which sometimes carry another type of erm gene, ermF. A 19-kbp segment of DNA from CTnGERM1 was cloned and sequenced. A 10-kbp portion of this segment hybridized not only to DNA from all the ermG-containing strains but also to DNA from strains that did not carry ermG. Thus, CTnGERM1 seems to be part of a family of CTns, some of which have acquired ermG. The percentage of G+C content of the ermG region was significantly lower than that of the chromosome of Bacteroides species-an indication that CTnGERM1 may have entered Bacteroides strains from some other bacterial genus. A survey of strains isolated before 1970 and after 1990 suggests that the CTnGERM1 type of CTn entered Bacteroides species relatively recently. One of the genes located upstream of ermG encoded a protein that had 85% amino acid sequence identity with a macrolide efflux pump, MefA, from Streptococcus pyogenes. Our having found >90% sequence identity of two upstream genes, including mefA, and the remnants of two transposon-carried genes downstream of ermG with genes found previously only in gram-positive bacteria raises the possibility that gram-positive bacteria could have been the origin of CTnGERM1. PMID- 12902249 TI - Curvularia haloperoxidase: antimicrobial activity and potential application as a surface disinfectant. AB - A presumed antimicrobial enzyme system, the Curvularia haloperoxidase system, was examined with the aim of evaluating its potential as a sanitizing agent. In the presence of hydrogen peroxide, Curvularia haloperoxidase facilitates the oxidation of halides, such as chloride, bromide, and iodide, to antimicrobial compounds. The Curvularia haloperoxidase system caused several-log-unit reductions in counts of bacteria (Pseudomonas spp., Escherichia coli, Serratia marcescens, Aeromonas salmonicida, Shewanella putrefaciens, Staphylococcus epidermidis, and Listeria monocytogenes), yeasts (Candida sp. and Rhodotorula sp.), and filamentous fungi (Aspergillus niger, Aspergillus tubigensis, Aspergillus versicolor, Fusarium oxysporum, Penicillium chrysogenum, and Penicillium paxilli) cultured in suspension. Also, bacteria adhering to the surfaces of contact lenses were killed. The numbers of S. marcescens and S. epidermidis cells adhering to contact lenses were reduced from 4.0 and 4.9 log CFU to 1.2 and 2.7 log CFU, respectively, after treatment with the Curvularia haloperoxidase system. The killing effect of the Curvularia haloperoxidase system was rapid, and 10(6) CFU of E. coli cells/ml were eliminated within 10 min of treatment. Furthermore, the antimicrobial effect was short lived, causing no antibacterial effect against E. coli 10 min after the system was mixed. Bovine serum albumin (1%) and alginate (1%) inhibited the antimicrobial activity of the Curvularia haloperoxidase system, whereas glucose and Tween 20 did not affect its activity. In conclusion, the Curvularia haloperoxidase system is an effective sanitizing system and has the potential for a vast range of applications, for instance, for disinfection of contact lenses or medical devices. PMID- 12902248 TI - Improved quantitative estimates of low environmental loading and sporadic periparturient shedding of Cryptosporidium parvum in adult beef cattle. AB - Our primary goal was to generate an accurate estimate of the daily environmental loading rate of Cryptosporidium parvum oocysts for adult beef cattle, using immunomagnetic separation coupled with direct immunofluorescence microscopy for a highly sensitive diagnostic assay. An additional goal was to measure the prevalence and intensity of fecal shedding of C. parvum oocysts in pre- and postparturient cows as an indicator of their potential to infect young calves. This diagnostic method could detect with a > or = 90% probability oocyst concentrations as low as 3.2 oocysts g of feces(-1), with a 54% probability of detecting just one oocyst g of feces(-1). Using this diagnostic method, the overall apparent prevalence of adult beef cattle testing positive for C. parvum was 7.1% (17 of 240), with 8.3 and 5.8% of cattle shedding oocysts during the pre and postcalving periods, respectively. The mean intensity of oocyst shedding for test-positive cattle was 3.38 oocysts g of feces(-1). The estimated environmental loading rate of C. parvum ranged from 3,900 to 9,200 oocysts cow(-1) day(-1), which is substantially less than a previous estimate of 1.7 x 10(5) oocysts cow( 1) day(-1) (range of 7.7 x 10(4) to 2.3 x 10(5) oocysts cow(-1) day(-1)) (B. Hoar, E. R. Atwill, and T. B. Farver, Quant. Microbiol. 2:21-36, 2000). Use of this highly sensitive assay functioned to detect a greater proportion of low intensity shedders in our population of cattle, which reduced the estimated mean intensity of shedding and thereby reduced the associated environmental loading rate compared to those of previous studies. PMID- 12902250 TI - Evaluation of the use of PCR and reverse transcriptase PCR for detection of pathogenic bacteria in biosolids from anaerobic digestors and aerobic composters. AB - A PCR-based method and a reverse transcriptase PCR (RT-PCR)-based method were developed for the detection of pathogenic bacteria in organic waste, using Salmonella spp., Listeria monocytogenes, Yersinia enterocolitica, and Staphylococcus aureus as model organisms. In seeded organic waste samples, detection limits of less than 10 cells per g of organic waste were achieved after one-step enrichment of bacteria, isolation, and purification of DNA or RNA before PCR or RT-PCR amplification. To test the reproducibility and reliability of the newly developed methods, 46 unseeded samples were collected from diverse aerobic (composting) facilities and anaerobic digestors and analyzed by both culture based classical and newly developed PCR-based procedures. No false-positive but some false-negative results were generated by the PCR- or RT-PCR-based methods after one-step enrichment when compared to the classical detection methods. The results indicated that the level of activity of the tested bacteria in unseeded samples was very low compared to that of freshly inoculated cells, preventing samples from reaching the cell density required for PCR-based detection after one step enrichment. However, for Salmonella spp., a distinct PCR product could be obtained for all 22 nonamended samples that tested positive for Salmonella spp. by the classical detection procedure when a selective two-step enrichment (20 h in peptone water at 37 degrees C and 24 h in Rappaport Vassiliadis medium at 43 degrees C) was performed prior to nucleic acid extraction and PCR. Hence, the classical procedure was shortened, since cell plating and further differentiation of isolated colonies can be omitted, substituted for by highly sensitive and reliable detection based on nucleic acid extraction and PCR. Similarly, 2 of the 22 samples in which Salmonella spp. were detected also tested positive for Listeria monocytogenes according to a two-step enrichment procedure followed by PCR, compared to 3 samples that tested positive when classical isolation procedures were followed. The study shows that selective two-step enrichment is useful when very low numbers of bacterial pathogens must be detected in organic waste materials, such as biosolids. There were no false-positive results derived from DNA of dead cells in the waste sample, suggesting that it is not necessary to perform RT-PCR analyses when PCR is combined with selective enrichment. Large numbers of added nontarget bacteria did not affect detection of Salmonella spp., L. monocytogenes, and Y. enterocolitica but increased the detection limit of Staphylococcus aureus from <10 to 10(4) CFU/g of organic waste. Overall, the detection methods developed using seeded organic waste samples from one waste treatment facility (WTF) needed to be modified for satisfactory detection of pathogens in samples from other WTFs, emphasizing the need for extensive field testing of laboratory-derived PCR protocols. A survey of 13 WTFs in Germany revealed that all facilities complied with the German Biowaste Ordinance, which mandates that the end product after anaerobic digestion or aerobic composting be free of Salmonella In addition, all biosolids were free of L. monocytogenes, Staphylococcus aureus, and Y. enterocolitica, as evidenced by both classical and PCR-based detection methods. PMID- 12902251 TI - Characterization of the corrinoid iron-sulfur protein tetrachloroethene reductive dehalogenase of Dehalobacter restrictus. AB - The membrane-bound tetrachloroethene reductive dehalogenase (PCE-RDase) (PceA; EC 1.97.1.8), the terminal component of the respiratory chain of Dehalobacter restrictus, was purified 25-fold to apparent electrophoretic homogeneity. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed a single band with an apparent molecular mass of 60 +/- 1 kDa, whereas the native molecular mass was 71 +/- 8 kDa according to size exclusion chromatography in the presence of the detergent octyl-beta-D-glucopyranoside. The monomeric enzyme contained (per mol of the 60-kDa subunit) 1.0 +/- 0.1 mol of cobalamin, 0.6 +/- 0.02 mol of cobalt, 7.1 +/- 0.6 mol of iron, and 5.8 +/- 0.5 mol of acid-labile sulfur. Purified PceA catalyzed the reductive dechlorination of tetrachloroethene and trichloroethene to cis-1,2-dichloroethene with a specific activity of 250 +/- 12 nkat/mg of protein. In addition, several chloroethanes and tetrachloromethane caused methyl viologen oxidation in the presence of PceA. The K(m) values for tetrachloroethene, trichloroethene, and methyl viologen were 20.4 +/- 3.2, 23.7 +/- 5.2, and 47 +/- 10 micro M, respectively. The PceA exhibited the highest activity at pH 8.1 and was oxygen sensitive, with a half-life of activity of 280 min upon exposure to air. Based on the almost identical N-terminal amino acid sequences of PceA of Dehalobacter restrictus, Desulfitobacterium hafniense strain TCE1 (formerly Desulfitobacterium frappieri strain TCE1), and Desulfitobacterium hafniense strain PCE-S (formerly Desulfitobacterium frappieri strain PCE-S), the pceA genes of the first two organisms were cloned and sequenced. Together with the pceA genes of Desulfitobacterium hafniense strains PCE-S and Y51, the pceA genes of Desulfitobacterium hafniense strain TCE1 and Dehalobacter restrictus form a coherent group of reductive dehalogenases with almost 100% sequence identity. Also, the pceB genes, which may code for a membrane anchor protein of PceA, and the intergenic regions of Dehalobacter restrictus and the three desulfitobacteria had identical sequences. Whereas the cprB (chlorophenol reductive dehalogenase) genes of chlorophenol-dehalorespiring bacteria are always located upstream of cprA, all pceB genes known so far are located downstream of pceA. The possible consequences of this feature for the annotation of putative reductive dehalogenase genes are discussed, as are the sequence around the iron sulfur cluster binding motifs and the type of iron-sulfur clusters of the reductive dehalogenases of Dehalobacter restrictus and Desulfitobacterium dehalogenans identified by electron paramagnetic resonance spectroscopy. PMID- 12902252 TI - Genetic diversity and temporal variation in the cyanophage community infecting marine Synechococcus species in Rhode Island's coastal waters. AB - The cyanophage community in Rhode Island's coastal waters is genetically diverse and dynamic. Cyanophage abundance ranged from over 10(4) phage ml(-1) in the summer months to less then 10(2) phage ml(-1) during the winter months. Thirty six distinct cyanomyovirus g20 genotypes were identified over a 3-year sampling period; however, only one to nine g20 genotypes were detected at any one sampling date. Phylogenetic analyses of g20 sequences revealed that the Rhode Island cyanomyoviral isolates fall into three main clades and are closely related to other known viral isolates of Synechococcus spp. Extinction dilution enrichment followed by host range tests and PCR restriction fragment length polymorphism analysis was used to detect changes in the relative abundance of cyanophage types in June, July, and August 2002. Temporal changes in both the overall composition of the cyanophage community and the relative abundance of specific cyanophage g20 genotypes were observed. In some seawater samples, the g20 gene from over 50% of isolated cyanophages could not be amplified by using the PCR primer pairs specific for cyanomyoviruses, which suggested that cyanophages in other viral families (e.g., Podoviridae or Siphoviridae) may be important components of the Rhode Island cyanophage community. PMID- 12902254 TI - Effect of incubation temperature on isolation of Campylobacter jejuni genotypes from foodstuffs enriched in Preston broth. AB - Preston broth and agar incubated at either 37 or 42 degrees C have been widely used to isolate campylobacters from foodstuffs. The consequences of using either incubation temperature were investigated. Retail packs of raw chicken (n = 24) and raw lamb liver (n = 30) were purchased. Samples were incubated in Preston broth at 37 and 42 degrees C and then streaked onto Preston agar and incubated as before. Two Campylobacter isolates per treatment were characterized. Poultry isolates were genotyped by random amplification of polymorphic DNA (RAPD), pulsed field gel electrophoresis (PFGE), and flagellin PCR-restriction fragment length polymorphism, and lamb isolates were genotyped by RAPD only. In total, 96% of the poultry and 73% of the lamb samples yielded campylobacters. The lamb isolates were all Campylobacter jejuni, as were 96% of the poultry isolates, with the remainder being Campylobacter lari. The incubation temperature had no significant effect on the number of positive samples or on the species isolated. However, genotyping of the C. jejuni isolates revealed profound differences in the types obtained. Overall (from poultry and lamb), the use of a single incubation temperature, 37 degrees C, gave 56% of the total number of RAPD C. jejuni genotypes, and hence, 44% remained undetected. The effect was especially marked in the poultry samples, where incubation at 37 degrees C gave 47% of the PFGE genotypes but 53% were exclusively recovered after incubation at 42 degrees C. Thus, the incubation temperature of Preston media selects for certain genotypes of C. jejuni, and to detect the widest range, samples should be incubated at both 37 and 42 degrees C. Conversely, genotyping results arising from the use of a single incubation temperature should be interpreted with caution. PMID- 12902253 TI - The mode of action of the Bacillus thuringiensis vegetative insecticidal protein Vip3A differs from that of Cry1Ab delta-endotoxin. AB - The Vip3A protein, secreted by Bacillus spp. during the vegetative stage of growth, represents a new family of insecticidal proteins. In our investigation of the mode of action of Vip3A, the 88-kDa Vip3A full-length toxin (Vip3A-F) was proteolytically activated to an approximately 62-kDa core toxin either by trypsin (Vip3A-T) or lepidopteran gut juice extracts (Vip3A-G). Biotinylated Vip3A-G demonstrated competitive binding to lepidopteran midgut brush border membrane vesicles (BBMV). Furthermore, in ligand blotting experiments with BBMV from the tobacco hornworm, Manduca sexta (Linnaeus), activated Cry1Ab bound to 120-kDa aminopeptidase N (APN)-like and 250-kDa cadherin-like molecules, whereas Vip3A-G bound to 80-kDa and 100-kDa molecules which are distinct from the known Cry1Ab receptors. In addition, separate blotting experiments with Vip3A-G did not show binding to isolated Cry1A receptors, such as M. sexta APN protein, or a cadherin Cry1Ab ecto-binding domain. In voltage clamping assays with dissected midgut from the susceptible insect, M. sexta, Vip3A-G clearly formed pores, whereas Vip3A-F was incapable of pore formation. In the same assay, Vip3A-G was incapable of forming pores with larvae of the nonsusceptible insect, monarch butterfly, Danaus plexippus (Linnaeus). In planar lipid bilayers, both Vip3A-G and Vip3A-T formed stable ion channels in the absence of any receptors, supporting pore formation as an inherent property of Vip3A. Both Cry1Ab and Vip3A channels were voltage independent and highly cation selective; however, they differed considerably in their principal conductance state and cation specificity. The mode of action of Vip3A supports its use as a novel insecticidal agent. PMID- 12902255 TI - PCR-restriction fragment length polymorphism method for detection of Cyclospora cayetanensis in environmental waters without microscopic confirmation. AB - We developed an alternative nested-PCR-restriction fragment length polymorphism (RFLP) protocol for the detection of Cyclospora cayetanensis in environmental samples that obviates the need for microscopic examination. The RFLP method, with the restriction enzyme AluI, differentiates the amplified target sequence from C. cayetanensis from those that may cross-react. This new protocol was used to reexamine a subset (121 of 180) of surface water samples. Samples previously positive when the CYCF3E and CYCR4B primers (33) and RFLP with MnlI (20) were used were also PCR positive with the new primers; however, they were RFLP negative. We verified, by sequencing these amplicons, that while two were most likely other Cyclospora species, they were not C. cayetanensis. We can detect as few as one oocyst seeded into an autoclaved pellet flocculated from 10 liters of surface water. This new protocol should be of great use for environmental microbiologists and public health laboratories. PMID- 12902256 TI - virF-positive Yersinia pseudotuberculosis and Yersinia enterocolitica found in migratory birds in Sweden. AB - During spring and autumn migrations, 468 fecal samples from 57 different species of migratory birds were collected in Sweden. In total, Yersinia spp. were isolated from 12.8% of collected samples. The most commonly found species was Yersinia enterocolitica, which was isolated from 5.6% of all collected samples, followed by Y. intermedia (3.8%), Y. frederiksenii (3.0%), Y. kristensenii (0.9%), Y. pseudotuberculosis (0.6%), and Y. rohdei (0.4%). The pathogenic, virF positive Y. pseudotuberculosis strains were recovered from three thrushes. These strains belonged to the same bioserotype, 1/O:2, but had two different profiles as determined by pulsed-field gel electrophoresis with NotI and SpeI enzymes. In addition, 10 Y. enterocolitica strains, all from barnacle geese, belonged to bioserotype 3/O:3, which is associated with human disease. Two of the strains were pathogenic, carrying the virF gene on their plasmids. All pathogenic Y. pseudotuberculosis and Y. enterocolitica strains were recovered during the spring, and as the birds were caught during active migration they likely became infected at an earlier stage of the migration, thus potentially transporting these bacterial pathogens over long geographical distances. PMID- 12902257 TI - Alternative splicing of transcripts from crtI and crtYB genes of Xanthophyllomyces dendrorhous. AB - Xanthophyllomyces dendrorhous is one of the relevant sources of the carotenoid astaxanthin. In this paper, we describe for the first time cloning of unexpected cDNAs obtained from the crtI and crtYB genes of X. dendrorhous strain UCD 67-385. The cDNA of the crtI gene conserves 80 bp of the first intron, while the cDNA of the crtYB gene conserves 55 bp of the first intron and lacks 111 bp of the second exon. The crtI and crtYB RNAs could be spliced in alternative splice sites, which produced alternative transcripts which could not be translated to active CRTI and CRTYB proteins since they had numerous stop codons in their sequences. The ratio of mature mRNA to alternative mRNA for the crtI gene decreased as a function of the age of the culture, while the cellular content of carotenoids increased. It is possible that splicing to mature or alternative transcripts could regulate the cellular concentrations of phytoene desaturase and phytoene synthase-lycopene cyclase proteins, depending on the physiological or environmental conditions. PMID- 12902258 TI - Characterization of Escherichia coli O157:H7 from downer and healthy dairy cattle in the upper Midwest region of the United States. AB - While cattle in general have been identified as a reservoir of Escherichia coli O157:H7, there are limited data regarding the prevalence and clonality of this pathogen in downer dairy cattle and the potential impact to human health that may occur following consumption of meat derived from downer dairy cattle. In the present study, conducted at two slaughter facilities in Wisconsin between May and October of 2001, we established a higher prevalence of E. coli O157:H7 in fecal and/or tissue samples obtained aseptically from intact colons of downer dairy cattle (10 of 203, 4.9%) than in those from healthy dairy cattle (3 of 201, 1.5%). Analyses of 57 isolates, representing these 13 positive samples (one to five isolates per sample), by pulsed-field gel electrophoresis, revealed 13 distinct XbaI restriction endonuclease digestion profiles (REDP). Typically, isolates from different animals displayed distinct REDP and isolates from the same fecal or colon sample displayed indistinguishable REDP. However, in one sample, two different, but highly related, REDP were displayed by the isolates recovered. Antimicrobial susceptibility testing indicated that 10 of the 57 isolates, recovered from 2 (1 downer and 1 healthy animal) of the 13 positive samples, were resistant to at least 1 of 18 antimicrobials tested. However, there was no appreciable difference in the frequency of resistance of isolates recovered from downer and healthy dairy cattle, and not all isolates with the same REDP displayed the same antimicrobial susceptibility profile. Lastly, it was not possible to distinguish between isolates recovered from downer and healthy cattle based on their XbaI REDP or antimicrobial susceptibility. These results indicate that downer cattle had a 3.3-fold-higher prevalence of E. coli O157:H7 than healthy cattle within the time frame and geographic scope of this study. PMID- 12902259 TI - Growth of Rhodosporidium toruloides strain DBVPG 6662 on dibenzothiophene crystals and orimulsion. AB - Strains DBVPG 6662 and DBVPG 6739 of Rhodosporidium toruloides, a basidiomycete yeast, grew on thiosulfate as a sulfur source and glucose (2 g liter(-1) or 10.75 mM) as a carbon source. DBVPG 6662 has a defective sulfate transport system, whereas DBVPG 6739 barely grew on sulfate. They were compared for the ability to use dibenzothiophene (DBT) and related organic sulfur compounds as sulfur sources. In the presence of glucose as a carbon source and DBT as a sulfur source, strain DBVPG 6662 grew better than DBVPG 6739. In the presence of thiosulfate as a sulfur source, the two yeast strains did not use DBT, DBT sulfone, benzenesulfonic acid, biphenyl, and fluorene. When the two strains were grown in the presence of glucose, strain DBVPG 6662 transformed 27% of the DBT present (10 micro M) at a rate of 0.023 micro mol liter(-1) h(-1) in 36 h. Traces of 2,2'-dihydroxylated biphenyl were transiently accumulated under these conditions. When the same strain was grown on glucose in the presence of a higher concentration of DBT (0.5 g liter(-1)), mainly in an insoluble form, the whole surface of the DBT crystals was colonized by a thick mycelium. This adherent structure was imaged by confocal microscopy with fluorescent concanavalin A, a lectin that specifically binds glucose and mannose residues. When DBVPG 6662 was grown on glucose in the presence of a commercial emulsion of bitumen, i.e., orimulsion, 68% of the benzo- and dibenzothiophenes and DBTs was removed after 15 days of incubation. The fungus adhered by hyphae to orimulsion droplets. When cultivated in the presence of commercial emulsifier-free fuel oil containing alkylated benzothiophenes and DBTs and having a composition similar to that of orimulsion, strain DBVPG 6662 removed only 11% of the total organic sulfur that occurs in the medium and did not adhere to the oil droplets. These results indicate that strain DBVPG 6662 is able to utilize the organic sulfur of DBT and a large variety of thiophenic compounds that occur extensively in commercial fuel oils by physically adhering to the organic sulfur source. PMID- 12902260 TI - Rapid detection of meat spoilage by measuring volatile organic compounds by using proton transfer reaction mass spectrometry. AB - The evolution of the microbial spoilage population for air- and vacuum-packaged meat (beef and pork) stored at 4 degrees C was investigated over 11 days. We monitored the viable counts (mesophilic total aerobic bacteria, Pseudomonas spp., Enterobacteriaceae, lactic acid bacteria, and Enterococcus spp.) by the microbiological standard technique and by measuring the emission of volatile organic compounds (VOCs) with the recently developed proton transfer reaction mass spectrometry system. Storage time, packaging type, and meat type had statistically significant (P < 0.05) effects on the development of the bacterial numbers. The concentrations of many of the measured VOCs, e.g., sulfur compounds, largely increased over the storage time. We also observed a large difference in the emissions between vacuum- and air-packaged meat. We found statistically significant strong correlations (up to 99%) between some of the VOCs and the bacterial contamination. The concentrations of these VOCs increased linearly with the bacterial numbers. This study is a first step toward replacing the time consuming plate counting by fast headspace air measurements, where the bacterial spoilage can be determined within minutes instead of days. PMID- 12902261 TI - Photobactin: a catechol siderophore produced by Photorhabdus luminescens, an entomopathogen mutually associated with Heterorhabditis bacteriophora NC1 nematodes. AB - The nematode Heterorhabditis bacteriophora transmits a monoculture of Photorhabdus luminescens bacteria to insect hosts, where it requires the bacteria for efficient insect pathogenicity and as a substrate for growth and reproduction. Siderophore production was implicated as being involved in the symbiosis because an ngrA mutant inadequate for supporting nematode growth and reproduction was also deficient in producing siderophore activity and ngrA is homologous to a siderophore biosynthetic gene, entD. The role of the siderophore in the symbiosis with the nematode was determined by isolating and characterizing a mini-Tn5-induced mutant, NS414, producing no detectable siderophore activity. This mutant, being defective for growth in iron-depleted medium, was normal in supporting nematode growth and reproduction, in transmission by the dauer juvenile nematode, and in insect pathogenicity. The mini-Tn5 transposon was inserted into phbH; whose protein product is a putative peptidyl carrier protein homologous to the nonribosomal peptide synthetase VibF of Vibrio cholerae. Other putative siderophore biosynthetic and transport genes flanking phbH were characterized. The catecholate siderophore was purified, its structure was determined to be 2-(2,3-dihydroxyphenyl)-5-methyl-4,5-dihydro-oxazole-4 carboxylic acid [4-(2,3-dihydroxybenzoylamino)-butyl]-amide, and it was given the generic name photobactin. Antibiotic activity was detected with purified photobactin, indicating that the siderophore may contribute to antibiosis of the insect cadaver. These results eliminate the lack of siderophore activity as the cause for the inadequacy of the ngrA mutant in supporting nematode growth and reproduction. PMID- 12902262 TI - Occurrence of Escherichia coli and enterococci in Cladophora (Chlorophyta) in nearshore water and beach sand of Lake Michigan. AB - Each summer, the nuisance green alga Cladophora (mostly Cladophora glomerata) amasses along Lake Michigan beaches, creating nearshore anoxia and unsightly, malodorous mats that can attract problem animals and detract from visitor enjoyment. Traditionally, elevated counts of Escherichia coli are presumed to indicate the presence of sewage, mostly derived from nearby point sources. The relationship between fecal indicator bacteria and Cladophora remains essentially unstudied. This investigation describes the local and regional density of Escherichia coli and enterococci in Cladophora mats along beaches in the four states (Wisconsin, Illinois, Indiana, and Michigan) bordering Lake Michigan. Samples of Cladophora strands collected from 10 beaches (n = 41) were assayed for concentrations of E. coli and enterococci during the summer of 2002. Both E. coli and enterococci were ubiquitous (up to 97% occurrence), with overall log mean densities (+/- standard errors) of 5.3 (+/- 4.8) and 4.8 (+/- 4.5) per g (dry weight). E. coli and enterococci were strongly correlated in southern Lake Michigan beaches (P < 0.001, R(2) = 0.73, n = 17) but not in northern beaches (P = 0.892, n = 16). Both E. coli and enterococci survived for over 6 months in sun dried Cladophora mats stored at 4 degrees C; the residual bacteria in the dried alga readily grew upon rehydration. These findings suggest that Cladophora amassing along the beaches of Lake Michigan may be an important environmental source of indicator bacteria and call into question the reliability of E. coli and enterococci as indicators of water quality for freshwater recreational beaches. PMID- 12902263 TI - Intraspecies polymorphism of Cryptosporidium parvum revealed by PCR-restriction fragment length polymorphism (RFLP) and RFLP-single-strand conformational polymorphism analyses. AB - A glycoprotein (Cpgp40/15)-encoding gene of Cryptosporidium parvum was analyzed to reveal intraspecies polymorphism within C. parvum isolates. Forty-one isolates were collected from different geographical origins (Japan, Italy, and Nepal) and hosts (humans, calves, and a goat). These isolates were characterized by means of DNA sequencing, PCR-restriction fragment length polymorphism (PCR-RFLP), and RFLP single-strand conformational polymorphism (RFLP-SSCP) analyses of the gene for Cpgp40/15. The sequence analysis indicated that there was DNA polymorphism between genotype I and II, as well as within genotype I, isolates. The DNA and amino acid sequence identities between genotypes I and II differed, depending on the isolates, ranging from 73.3 to 82.9% and 62.4 to 80.1%, respectively. Those among genotype I isolates differed, depending on the isolates, ranging from 69.0 to 85.4% and 54.8 to 79.2%, respectively. Because of the high resolution generated by PCR-RFLP and RFLP-SSCP, the isolates of genotype I could be subtyped as genotypes Ia1, Ia2, Ib, and Ie. The isolates of genotype II could be subtyped as genotypes IIa, IIb, and IIc. The isolates from calves, a goat, and one Japanese human were identified as genotype II. Within genotype II, the isolates from Japan were identified as genotype IIa, those from calves in Italy were identified as genotype IIb, and the goat isolate was identified as genotype IIc. All of the genotype I isolates were from humans. The Japanese isolate (code no. HJ3) and all of the Nepalese isolates were identified as genotypes Ia1 and Ia2, respectively. The Italian isolates were identified as genotype Ib, and the Japanese isolate (code no. HJ2) was identified as genotype Ie. Thus, the PCR-RFLP SSCP analysis of this glycoprotein Cpgp40/15 gene generated a high resolution that has not been achieved by previous methods of genotypic differentiation of C. parvum. PMID- 12902264 TI - A cytochrome c from a lupanine-transforming Pseudomonas putida strain is expressed in Escherichia coli during aerobic cultivation and efficiently exported and assembled in the periplasm. AB - We have cloned, sequenced, and heterologously expressed a periplasmic cytochrome c from a lupanine-utilizing Pseudomonas putida strain. Aerobic batch cultivation of Escherichia coli TB1 harboring the cytochrome c gene placed downstream of the lac promoter in pUC9 vector resulted in significant production of the holo cytochrome c in the periplasm ( approximately 4 mg of hemoprotein/liter of culture). The recombinant cytochrome c was purified to homogeneity and was found to be functional in accepting electrons from lupanine hydroxylase while catalyzing hydroxylation of lupanine. Comparison of the N-terminal amino acid sequence of the isolated cytochrome c with that deduced from the DNA sequence indicated that the signal sequence was processed at the bond position predicted by the SigPep program. The molecular size of the cytochrome c determined by electrospray mass spectrometry (9,595) was in precise agreement with that predicted from the nucleotide sequence. PMID- 12902265 TI - Metabolic engineering of ammonium assimilation in xylose-fermenting Saccharomyces cerevisiae improves ethanol production. AB - Cofactor imbalance impedes xylose assimilation in Saccharomyces cerevisiae that has been metabolically engineered for xylose utilization. To improve cofactor use, we modified ammonia assimilation in recombinant S. cerevisiae by deleting GDH1, which encodes an NADPH-dependent glutamate dehydrogenase, and by overexpressing either GDH2, which encodes an NADH-dependent glutamate dehydrogenase, or GLT1 and GLN1, which encode the GS-GOGAT complex. Overexpression of GDH2 increased ethanol yield from 0.43 to 0.51 mol of carbon (Cmol) Cmol(-1), mainly by reducing xylitol excretion by 44%. Overexpression of the GS-GOGAT complex did not improve conversion of xylose to ethanol during batch cultivation, but it increased ethanol yield by 16% in carbon-limited continuous cultivation at a low dilution rate. PMID- 12902266 TI - Enhanced production of insulin-like growth factor I fusion protein in Escherichia coli by coexpression of the down-regulated genes identified by transcriptome profiling. AB - The transcriptome profiles of recombinant Escherichia coli producing human insulin-like growth factor I fusion protein (IGF-I(f)) during the high-cell density fed-batch culture were analyzed using DNA microarrays. The expression levels of 529 genes were significantly altered after induction. About 200 genes were significantly down-regulated during the production of IGF-I(f) after induction. Among these down-regulated genes, we rationally selected and coexpressed in E. coli producing IGF-I(f) the prsA gene (encoding a phosphoribosyl pyrophosphate synthetase) and the glpF gene (encoding a glycerol transporter), which are involved in an early key step in the biosynthetic pathway of nucleotides and amino acids (Trp and His) and the first step in glycerol utilization, respectively. As a result, the production of IGF-I(f) could be increased from 1.8 +/- 0.13 (+/- standard deviation) to 4.3 +/- 0.24 g/liter. The volumetric productivity was also increased from 0.36 +/- 0.027 to 0.82 +/- 0.048 g/liter/h. These results demonstrate that transcriptome profiling can provide invaluable information in designing engineered strains showing enhanced performance. PMID- 12902268 TI - Real-time PCR detection of Brucella abortus: a comparative study of SYBR green I, 5'-exonuclease, and hybridization probe assays. AB - Real-time PCR provides a means of detecting and quantifying DNA targets by monitoring PCR product accumulation during cycling as indicated by increased fluorescence. A number of different approaches can be used to generate the fluorescence signal. Three approaches-SYBR Green I (a double-stranded DNA intercalating dye), 5'-exonuclease (enzymatically released fluors), and hybridization probes (fluorescence resonance energy transfer)-were evaluated for use in a real-time PCR assay to detect Brucella abortus. The three assays utilized the same amplification primers to produce an identical amplicon. This amplicon spans a region of the B. abortus genome that includes portions of the alkB gene and the IS711 insertion element. All three assays were of comparable sensitivity, providing a linear assay over 7 orders of magnitude (from 7.5 ng down to 7.5 fg). However, the greatest specificity was achieved with the hybridization probe assay. PMID- 12902267 TI - An in vitro study of the probiotic potential of a bile-salt-hydrolyzing Lactobacillus fermentum strain, and determination of its cholesterol-lowering properties. AB - This study evaluated the use of a bile-salt-hydrolyzing Lactobacillus fermentum strain as a probiotic with potential hypocholesterolemic properties. The effect of L. fermentum on representative microbial populations and overall metabolic activity of the human intestinal microbiota was investigated using a three-stage continuous culture system. Also, the use of galactooligosaccharides as a prebiotic to enhance growth and/or activity of the Lactobacillus strain was evaluated. Administration of L. fermentum resulted in a decrease in the overall bifidobacterial population (ca. 1 log unit). In the in vitro system, no significant changes were observed in the total bacterial, Lactobacillus, Bacteroides, and clostridial populations through L. fermentum supplementation. Acetate production decreased by 9 to 27%, while the propionate and butyrate concentrations increased considerably (50 to 90% and 52 to 157%, respectively). A general, although lesser, increase in the production of lactate was observed with the administration of the L. fermentum strain. Supplementation of the prebiotic to the culture medium did not cause statistically significant changes in either the numbers or the activity of the microbiota, although an increase in the butyrate production was seen (29 to 39%). Results from this in vitro study suggest that L. fermentum KC5b is a candidate probiotic which may affect cholesterol metabolism. The short-chain fatty acid concentrations, specifically the molar proportion of propionate and/or bile salt deconjugation, are probably the major mechanism involved in the purported cholesterol-lowering properties of this strain. PMID- 12902269 TI - Characterization of Streptococcus mutans strains deficient in EIIAB Man of the sugar phosphotransferase system. AB - The phosphoenolpyruvate:sugar phosphotransferase system (PTS) is the major sugar uptake system in oral streptococci. The role of EIIAB(Man) (encoded by manL) in gene regulation and sugar transport was investigated in Streptococcus mutans UA159. The manL knockout strain, JAM1, grew more slowly than the wild-type strain in glucose but grew faster in mannose and did not display diauxic growth, indicating that EIIAB(Man) is involved in sugar uptake and in carbohydrate catabolite repression. PTS assays of JAM1, and of strains lacking the inducible (fruI) and constitutive (fruCD) EII fructose, revealed that S. mutans EIIAB(Man) transported mannose and glucose and provided evidence that there was also a mannose-inducible or glucose-repressible mannose PTS. Additionally, there appears to be a fructose PTS that is different than FruI and FruCD. To determine whether EIIAB(Man) controlled expression of the known virulence genes, glucosyltransferases (gtfBC) and fructosyltransferase (ftf) promoter fusions of these genes were established in the wild-type and EIIAB(Man)-deficient strains. In the manL mutant, the level of chloramphenicol acetyltransferase activity expressed from the gtfBC promoter was up to threefold lower than that seen with the wild-type strain at pH 6 and 7, indicating that EIIAB(Man) is required for optimal expression of gtfBC. No significant differences were observed between the mutant and the wild-type background in ftf regulation, with the exception that under glucose-limiting conditions at pH 7, the mutant exhibited a 2.1-fold increase in ftf expression. Two-dimensional gel analysis of batch-grown cells of the EIIAB(Man)-deficient strain indicated that the expression of at least 38 proteins was altered compared to that seen with the wild-type strain, revealing that EIIAB(Man) has a pleiotropic effect on gene expression. PMID- 12902270 TI - Effects of a chlorhexidine gluconate-containing mouthwash on the vitality and antimicrobial susceptibility of in vitro oral bacterial ecosystems. AB - Oral bacterial microcosms, established using saliva inocula from three individuals, were maintained under a feast-famine regime within constant-depth film fermenters. Steady-state communities were exposed four times daily, postfeeding, to a chlorhexidine (CHX) gluconate-containing mouthwash (CHXM) diluted to 0.06% (wt/vol) antimicrobial content. The microcosms were characterized by heterotrophic plate counts and PCR-denaturing gradient gel electrophoresis (DGGE). CHXM caused significant decreases in both total anaerobe and total aerobe/facultative anaerobe counts (P < 0.05), together with lesser decreases in gram-negative anaerobes. The degree of streptococcal and actinomycete inhibition varied considerably among individuals. DGGE showed that CHXM exposure caused considerable decreases in microbial diversity, including marked reductions in Prevotella sp. and Selenomonas infelix. Pure-culture studies of 10 oral bacteria (eight genera) showed that Actinomyces naeslundii, Veillonella dispar, Prevotella nigrescens, and the streptococci were highly susceptible to CHX, while Lactobacillus rhamnosus, Fusobacterium nucleatum, and Neisseria subflava were the least susceptible. Determination of the MICs of triclosan, CHX, erythromycin, penicillin V, vancomycin, and metronidazole for microcosm isolates, before and after 5 days of CHXM exposure, showed that CHXM exposure altered the distribution of isolates toward those that were less susceptible to CHX (P < 0.05). Changes in susceptibility distributions for the other test agents were not statistically significant. In conclusion, population changes in plaque microcosms following repeated exposure to CHXM represented an inhibition of the most susceptible flora with a clonal expansion of less susceptible species. PMID- 12902271 TI - Yeast genome-wide expression analysis identifies a strong ergosterol and oxidative stress response during the initial stages of an industrial lager fermentation. AB - Genome-wide expression analysis of an industrial strain of Saccharomyces cerevisiae during the initial stages of an industrial lager fermentation identified a strong response from genes involved in the biosynthesis of ergosterol and oxidative stress protection. The induction of the ERG genes was confirmed by Northern analysis and was found to be complemented by a rapid accumulation of ergosterol over the initial 6-h fermentation period. From a test of the metabolic activity of deletion mutants in the ergosterol biosynthesis pathway, it was found that ergosterol is an important factor in restoring the fermentative capacity of the cell after storage. Additionally, similar ERG10 and TRR1 gene expression patterns over the initial 24-h fermentation period highlighted a possible interaction between ergosterol biosynthesis and the oxidative stress response. Further analysis showed that erg mutants producing altered sterols were highly sensitive to oxidative stress-generating compounds. Here we show that genome-wide expression analysis can be used in the commercial environment and was successful in identifying environmental conditions that are important in industrial yeast fermentation. PMID- 12902272 TI - Detection and quantification of Plectosphaerella cucumerina, a potential biological control agent of potato cyst nematodes, by using conventional PCR, real-time PCR, selective media, and baiting. AB - Potato cyst nematodes (PCN) are serious pests in commercial potato production, causing yield losses valued at approximately $300 million in the European Community. The nematophagous fungus Plectosphaerella cucumerina has demonstrated its potential as a biological control agent against PCN populations by reducing field populations by up to 60% in trials. The use of biological control agents in the field requires the development of specific techniques to monitor the release, population size, spread or decline, and pathogenicity against its host. A range of methods have therefore been developed to monitor P. cucumerina. A species specific PCR primer set (PcCF1-PcCR1) was designed that was able to detect the presence of P. cucumerina in soil, root, and nematode samples. PCR was combined with a bait method to identify P. cucumerina from infected nematode eggs, confirming the parasitic ability of the fungus. A selective medium was adapted to isolate the fungus from root and soil samples and was used to quantify the fungus from field sites. A second P. cucumerina-specific primer set (PcRTF1-PcRTR1) and a Taqman probe (PcRTP1) were designed for real-time PCR quantification of the fungus and provided a very sensitive means of detecting the fungus from soil. PCR, bait, and culture methods were combined to investigate the presence and abundance of P. cucumerina from two field sites in the United Kingdom where PCN populations were naturally declining. All methods enabled differences in the activity of P. cucumerina to be detected, and the results demonstrated the importance of using a combination of methods to investigate population size and activity of fungi. PMID- 12902273 TI - CynD, the cyanide dihydratase from Bacillus pumilus: gene cloning and structural studies. AB - The cyanide dihydratase in Bacillus pumilus was shown to be an 18-subunit spiral structure by three-dimensional reconstruction of electron micrographs of negatively stained material at its optimum pH, 8.0. At pH 5.4, the subunits rearrange to form an extended left-handed helix. Gel electrophoresis of glutaraldehyde cross-linked enzyme suggests that the fundamental component of the spiral is a dimer of the 37-kDa subunit. The gene was cloned, and the recombinant enzyme was readily expressed at high levels in Escherichia coli. Purification of the recombinant enzyme was facilitated by the addition of a C-terminal six histidine affinity purification tag. The tagged recombinant enzyme has K(m) and V(max) values similar to those published for the native enzyme. This is the first cyanide dihydratase from a gram-positive bacterium to be sequenced, and it is the first description of the structure of any member of this enzyme class. The putative amino acid sequence shares over 80% identity to the only other sequenced cyanide dihydratase, that of the gram-negative Pseudomonas stutzeri strain AK61, and is similar to a number of other bacterial and fungal nitrilases. This sequence similarity suggests that the novel short spiral structure may be typical of these enzymes. In addition, an active cyanide dihydratase from a non-cyanide degrading isolate of B. pumilus (strain 8A3) was cloned and expressed. This suggests that cynD, the gene coding for the cyanide dihydratase, is not unique to the C1 strain of B. pumilus and is not a reflection of its origin at a mining waste site. PMID- 12902274 TI - Targeting single-nucleotide polymorphisms in the 18S rRNA gene to differentiate Cyclospora species from Eimeria species by multiplex PCR. AB - Cyclospora cayetanensis is a coccidian parasite that causes protracted diarrheal illness in humans. C. cayetanensis is the only species of this genus thus far associated with human illness, although Cyclospora species from other primates have been named. The current method to detect the parasite uses a nested PCR assay to amplify a 294-bp region of the small subunit rRNA gene, followed by restriction fragment length polymorphism (RFLP) or DNA sequence analysis. Since the amplicons generated from C. cayetanensis and Eimeria species are the same size, the latter step is required to distinguish between these different species. The current PCR-RFLP protocol, however, cannot distinguish between C. cayetanensis and these new isolates. The differential identification of such pathogenic and nonpathogenic parasites is essential in assessing the risks to human health from microorganisms that may be potential contaminants in food and water sources. Therefore, to expand the utility of PCR to detect and identify these parasites in a multiplex assay, a series of genus- and species-specific forward primers were designed that are able to distinguish sites of limited sequence heterogeneity in the target gene. The most effective of these unique primers were those that identified single-nucleotide polymorphisms (SNPs) at the 3' end of the primer. Under more stringent annealing and elongation conditions, these SNP primers were able to differentiate between C. cayetanensis, nonhuman primate species of Cyclospora, and Eimeria species. As a diagnostic tool, the SNP PCR protocol described here presents a more rapid and sensitive alternative to the currently available PCR-RFLP detection method. In addition, the specificity of these diagnostic primers removes the uncertainty that can be associated with analyses of foods or environmental sources suspected of harboring potential human parasitic pathogens. PMID- 12902275 TI - N-acetyl-L-cysteine affects growth, extracellular polysaccharide production, and bacterial biofilm formation on solid surfaces. AB - N-Acetyl-L-cysteine (NAC) is used in medical treatment of patients with chronic bronchitis. The positive effects of NAC treatment have primarily been attributed to the mucus-dissolving properties of NAC, as well as its ability to decrease biofilm formation, which reduces bacterial infections. Our results suggest that NAC also may be an interesting candidate for use as an agent to reduce and prevent biofilm formation on stainless steel surfaces in environments typical of paper mill plants. Using 10 different bacterial strains isolated from a paper mill, we found that the mode of action of NAC is chemical, as well as biological, in the case of bacterial adhesion to stainless steel surfaces. The initial adhesion of bacteria is dependent on the wettability of the substratum. NAC was shown to bind to stainless steel, increasing the wettability of the surface. Moreover, NAC decreased bacterial adhesion and even detached bacteria that were adhering to stainless steel surfaces. Growth of various bacteria, as monocultures or in a multispecies community, was inhibited at different concentrations of NAC. We also found that there was no detectable degradation of extracellular polysaccharides (EPS) by NAC, indicating that NAC reduced the production of EPS, in most bacteria tested, even at concentrations at which growth was not affected. Altogether, the presence of NAC changes the texture of the biofilm formed and makes NAC an interesting candidate for use as a general inhibitor of formation of bacterial biofilms on stainless steel surfaces. PMID- 12902276 TI - Fidelity of select restriction endonucleases in determining microbial diversity by terminal-restriction fragment length polymorphism. AB - An evaluation of 18 DNA restriction endonucleases for use in terminal-restriction fragment length polymorphism (T-RFLP) analysis was performed by using richness and density indices in conjunction with computer simulations for 4,603 bacterial small-subunit rRNA gene sequences. T-RFLP analysis has become a commonly used method for screening environmental samples for precursory identification and community comparison studies due to its precision and high-throughput capability. The accuracy of T-RFLP analysis for describing a community has not yet been thoroughly evaluated. In this study, we attempted to classify restriction endonucleases based upon the ability to resolve unique terminal-restriction fragments (T-RFs) or operational taxonomic units (OTUs) from a database of gene sequences. Furthermore, we assessed the predictive accuracy of T-RFLP at fixed values of community richness (n = 1, 5, 10, 50, and 100). Classification of restriction endonuclease fidelity was performed by measuring richness and density for the entire database of T-RFs. Further analysis of T-RFLP accuracy for determining richness was performed by iterative, random sampling from the derived database of T-RFs. It became apparent that two constraints were influential for measuring the fidelity of a given restriction endonuclease: (i) the ability to resolve unique sequence variants and (ii) the number of unique T-RFs that fell within a measurable size range. The latter constraint was found to be more significant for estimating restriction endonuclease fidelity. Of the 18 restriction endonucleases examined, BstUI, DdeI, Sau96I, and MspI had the highest frequency of resolving single populations in model communities. All restriction endonucleases used in this study detected < or =70% of the OTUs at richness values greater than 50 OTUs per modeled community. Based on the results of our in silico experiments, the most efficacious uses of T-RFLP for microbial diversity studies are those that address situations where there is low to intermediate species richness (e.g., colonization, early successional stages, biofilm formation). PMID- 12902277 TI - Alteration of chain length substrate specificity of Aeromonas caviae R-enantiomer specific enoyl-coenzyme A hydratase through site-directed mutagenesis. AB - Aeromonas caviae R-specific enoyl-coenzyme A (enoyl-CoA) hydratase (PhaJ(Ac)) is capable of providing (R)-3-hydroxyacyl-CoA with a chain length of four to six carbon atoms from the fatty acid beta-oxidation pathway for polyhydroxyalkanoate (PHA) synthesis. In this study, amino acid substitutions were introduced into PhaJ(Ac) by site-directed mutagenesis to investigate the feasibility of altering the specificity for the acyl chain length of the substrate. A crystallographic structure analysis of PhaJ(Ac) revealed that Ser-62, Leu-65, and Val-130 define the width and depth of the acyl-chain-binding pocket. Accordingly, we targeted these three residues for amino acid substitution. Nine single-mutation enzymes and two double-mutation enzymes were generated, and their hydratase activities were assayed in vitro by using trans-2-octenoyl-CoA (C(8)) as a substrate. Three of these mutant enzymes, L65A, L65G, and V130G, exhibited significantly high activities toward octenoyl-CoA than the wild-type enzyme exhibited. PHA formation from dodecanoate (C(12)) was examined by using the mutated PhaJ(Ac) as a monomer supplier in recombinant Escherichia coli LS5218 harboring a PHA synthase gene from Pseudomonas sp. strain 61-3 (phaC1(Ps)). When L65A, L65G, or V130G was used individually, increased molar fractions of 3-hydroxyoctanoate (C(8)) and 3 hydroxydecanoate (C(10)) units were incorporated into PHA. These results revealed that Leu-65 and Val-130 affect the acyl chain length substrate specificity. Furthermore, comparative kinetic analyses of the wild-type enzyme and the L65A and V130G mutants were performed, and the mechanisms underlying changes in substrate specificity are discussed. PMID- 12902278 TI - The biphenyl- and 4-chlorobiphenyl-catabolic transposon Tn4371, a member of a new family of genomic islands related to IncP and Ti plasmids. AB - The nucleotide sequence of the biphenyl catabolic transposon Tn4371 has been completed and analyzed. It confirmed that the element has a mosaic structure made of several building blocks. In addition to previously identified genes coding for a tyrosine recombinase related to phage integrases and for biphenyl degradation enzymes very similar to those of Achromobacter georgiopolitanum KKS102, Tn4371 carries many plasmid-related genes involved in replication, partition, and other, as-yet-unknown, plasmid functions. One gene cluster contains most of the genes required to express a type IV secretion-mating pair formation apparatus coupled with a TraG ATPase, all of which are related to those found on IncP and Ti plasmids. Orthologues of all Tn4371 plasmid-related genes and of the tyrosine recombinase gene were found, with a very similar organization, in the chromosome of Ralstonia solanacearum and on the yet-to-be-determined genomic sequences of Erwinia chrysanthemi and Azotobacter vinelandii. In each of these chromosomal segments, conserved segments were separated by different groups of genes, which also differed from the Tn4371 bph genes. The conserved blocks of genes were also identified, in at least two copies, in the chromosome of Ralstonia metallidurans CH34. Tn4371 thus appears to represent a new family of potentially mobile genomic islands with a broad host range since they reside in a wide range of soil proteobacteria, including plant pathogens. PMID- 12902279 TI - Dual labeling of Pseudomonas putida with fluorescent proteins for in situ monitoring of conjugal transfer of the TOL plasmid. AB - We describe here a dual-labeling technique involving the green fluorescent protein (GFP) and the red fluorescent protein (DsRed) for in situ monitoring of horizontal gene transfer via conjugation. A GFPmut3b-tagged derivative of narrow host-range TOL plasmid (pWWO) was delivered to Pseudomonas putida KT2442, which was chromosomally labeled with dsRed by transposon insertion via biparental mating. Green and red fluorescent proteins were coexpressed in donor P. putida cells. Cells expressing both fluorescent proteins were smaller in size than cells expressing GFP alone. Donors and transconjugants in mixed culture or sludge samples were discriminated on the basis of their fluorescence by using confocal laser scanning microscopy. Conjugal plasmid transfer frequencies on agar surfaces and in sludge microcosms were determined microscopically without cultivation. This method worked well for in situ monitoring of horizontal gene transfer in addition to tracking the fate of microorganisms released into complex environments. To the best of our knowledge, this is the first study that discusses the coexpression of GFP and DsRed for conjugal gene transfer studies. PMID- 12902280 TI - Microbial ecology of an extreme acidic environment, the Tinto River. AB - The Tinto River (Huelva, southwestern Spain) is an extreme environment with a rather constant acidic pH along the entire river and a high concentration of heavy metals. The extreme conditions of the Tinto ecosystem are generated by the metabolic activity of chemolithotrophic microorganisms thriving in the rich complex sulfides of the Iberian Pyrite Belt. Molecular ecology techniques were used to analyze the diversity of this microbial community. The community's composition was studied by denaturing gradient gel electrophoresis (DGGE) using 16S rRNA and by 16S rRNA gene amplification. A good correlation between the two approaches was found. Comparative sequence analysis of DGGE bands showed the presence of organisms related to Leptospirillum spp., Acidithiobacillus ferrooxidans, Acidiphilium spp., "Ferrimicrobium acidiphilum," Ferroplasma acidiphilum, and Thermoplasma acidophilum. The different phylogenetic groups were quantified by fluorescent in situ hybridization with a set of rRNA-targeted oligonucleotide probes. More than 80% of the cells were affiliated with the domain Bacteria, with only a minor fraction corresponding to Archaea. Members of Leptospirillum ferrooxidans, Acidithiobacillus ferrooxidans, and Acidiphilium spp., all related to the iron cycle, accounted for most of the prokaryotic microorganisms detected. Different isolates of these microorganisms were obtained from the Tinto ecosystem, and their physiological properties were determined. Given the physicochemical characteristics of the habitat and the physiological properties and relative concentrations of the different prokaryotes found in the river, a model for the Tinto ecosystem based on the iron cycle is suggested. PMID- 12902281 TI - Directed evolution of Thermus maltogenic amylase toward enhanced thermal resistance. AB - The thermostability of maltogenic amylase from Thermus sp. strain IM6501 (ThMA) was improved greatly by random mutagenesis using DNA shuffling. Four rounds of DNA shuffling and subsequent recombination of the mutations produced the highly thermostable mutant enzyme ThMA-DM, which had a total of seven individual mutations. The seven amino acid substitutions in ThMA-DM were identified as R26Q, S169N, I333V, M375T, A398V, Q411L, and P453L. The optimal reaction temperature of the recombinant enzyme was 75 degrees C, which was 15 degrees C higher than that of wild-type ThMA, and the melting temperature, as determined by differential scanning calorimetry, was increased by 10.9 degrees C. The half-life of ThMA-DM was 172 min at 80 degrees C, a temperature at which wild-type ThMA was completely inactivated in less than 1 min. Six mutations that were generated during the evolutionary process did not significantly affect the specific activity of the enzyme, while the M375T mutation decreased activity to 23% of the wild-type level. The molecular interactions of the seven mutant residues that contributed to the increased thermostability of the mutant enzyme with other adjacent residues were examined by comparing the modeled tertiary structure of ThMA-DM with those of wild-type ThMA and related enzymes. The A398V and Q411L substitutions appeared to stabilize the enzyme by enhancing the interdomain hydrophobic interactions. The R26Q and P453L substitutions led potentially to the formation of genuine hydrogen bonds. M375T, which was located near the active site of ThMA, probably caused a conformational or dynamic change that enhanced thermostability but reduced the specific activity of the enzyme. PMID- 12902283 TI - Influence of seasonal environmental variables on the distribution of presumptive fecal Coliforms around an Antarctic research station. AB - Factors affecting fecal microorganism survival and distribution in the Antarctic marine environment include solar radiation, water salinity, temperature, sea ice conditions, and fecal input by humans and local wildlife populations. This study assessed the influence of these factors on the distribution of presumptive fecal coliforms around Rothera Point, Adelaide Island, Antarctic Peninsula during the austral summer and winter of February 1999 to September 1999. Each factor had a different degree of influence depending on the time of year. In summer (February), although the station population was high, presumptive fecal coliform concentrations were low, probably due to the biologically damaging effects of solar radiation. However, summer algal blooms reduced penetration of solar radiation into the water column. By early winter (April), fecal coliform concentrations were high, due to increased fecal input by migrant wildlife, while solar radiation doses were low. By late winter (September), fecal coliform concentrations were high near the station sewage outfall, as sea ice formation limited solar radiation penetration into the sea and prevented wind-driven water circulation near the outfall. During this study, environmental factors masked the effect of station population numbers on sewage plume size. If sewage production increases throughout the Antarctic, environmental factors may become less significant and effective sewage waste management will become increasingly important. These findings highlight the need for year-round monitoring of fecal coliform distribution in Antarctic waters near research stations to produce realistic evaluations of sewage pollution persistence and dispersal. PMID- 12902282 TI - High-level production of porphyrins in metabolically engineered Escherichia coli: systematic extension of a pathway assembled from overexpressed genes involved in heme biosynthesis. AB - Due to their spectroscopic properties porphyrins are of special interest for a variety of applications, ranging from drug development or targeting to material sciences and chemical and biological sensors. Since chemical syntheses are limited in terms of regio- and stereoselective functionalization of porphyrins, a biosynthetic approach with tailored enzyme catalysts offers a promising alternative. In this paper, we describe assembly of the entire heme biosynthetic pathway in a three-plasmid system and overexpression of the corresponding genes with Escherichia coli as a host. Without further optimization, this approach yielded remarkable porphyrin production levels, up to 90 micro mol/liter, which is close to industrial vitamin B(12) production levels. Different combinations of the genes were used to produce all major porphyrins that occur as intermediates in heme biosynthesis. All these porphyrin intermediates were obtained in high yields. The product spectrum was analyzed and quantified by using high performance liquid chromatography. Intriguingly, although protoporphyrin IX could be produced at high levels, overexpressed Bacillus subtilis ferrochelatase could not convert this substrate appreciably into heme. However, further investigation clearly revealed a high level of expression of the ferrochelatase and a high level of activity in vitro. These results may indicate that heme has a regulatory impact on the iron uptake of E. coli or that the ferrochelatase is inactive in vivo due to an incompatible enzyme interaction. PMID- 12902284 TI - Single-leaf resolution of the temporal population dynamics of Aureobasidium pullulans on apple leaves. AB - The abundance of phylloplane microorganisms typically varies over several orders of magnitude among leaves sampled concurrently. Because the methods traditionally used to sample leaves are destructive, it has remained unclear whether this high variability is due to fixed differences in habitat quality among leaves or to asynchronous temporal variation in the microbial population density on individual leaves. We developed a novel semidestructive assay to repeatedly sample the same apple leaves from orchard trees over time by removing progressively more proximal approximately 1-cm-wide transverse segments. Aureobasidium pullulans densities were determined by standard leaf homogenization and plating procedures and were expressed as CFU per square centimeter of segment. The A. pullulans population densities among leaves were lognormally distributed. The variability in A. pullulans population densities among subsections of a given leaf was one-third to one-ninth the variability among whole leaves harvested concurrently. Sequential harvesting of leaf segments did not result in detectable changes in A. pullulans density on residual leaf surfaces. These findings implied that we could infer whole-leaf A. pullulans densities over time by using partial leaves. When this successive sampling regimen was applied over the course of multiple 7- to 8-day experiments, the among-leaf effects were virtually always the predominant source of variance in A. pullulans density estimates. Changes in A. pullulans density tended to be synchronous among leaves, such that the rank order of leaves arrayed with respect to A. pullulans density was largely maintained through time. Occasional periods of asynchrony were observed, but idiosyncratic changes in A. pullulans density did not contribute appreciably to variation in the distribution of populations among leaves. This suggests that persistent differences in habitat (leaf) quality are primarily responsible for the variation in A. pullulans density among leaves in nature. PMID- 12902285 TI - Strain-specific ureolytic microbial calcium carbonate precipitation. AB - During a study of ureolytic microbial calcium carbonate (CaCO(3)) precipitation by bacterial isolates collected from different environmental samples, morphological differences were observed in the large CaCO(3) crystal aggregates precipitated within bacterial colonies grown on agar. Based on these differences, 12 isolates were selected for further study. We hypothesized that the striking differences in crystal morphology were the result of different microbial species or, alternatively, differences in the functional attributes of the isolates selected. Sequencing of 16S rRNA genes showed that all of the isolates were phylogenetically closely related to the Bacillus sphaericus group. Urease gene diversity among the isolates was examined by using a novel application of PCR denaturing gradient gel electrophoresis (DGGE). This approach revealed significant differences between the isolates. Moreover, for several isolates, multiple bands appeared on the DGGE gels, suggesting the apparent presence of different urease genes in these isolates. The substrate affinities (K(m)) and maximum hydrolysis rates (V(max)) of crude enzyme extracts differed considerably for the different strains. For certain isolates, the urease activity increased up to 10-fold in the presence of 30 mM calcium, and apparently this contributed to the characteristic crystal formation by these isolates. We show that strain specific calcification occurred during ureolytic microbial carbonate precipitation. The specificity was mainly due to differences in urease expression and the response to calcium. PMID- 12902286 TI - Remarkable diversity of phototrophic purple bacteria in a permanently frozen Antarctic lake. AB - Although anoxygenic photosynthesis is thought to play an important role in the primary productivity of permanently frozen lakes in the Antarctic dry valleys, the bacterial communities responsible for this metabolism remain uncharacterized. Here we report the composition and activity of phototrophic purple bacteria in Lake Fryxell, Antarctica, as determined by analysis of a photosynthesis-specific gene, pufM. The results revealed an extensive diversity and highly stratified distribution of purple nonsulfur bacteria in Lake Fryxell and showed which phylotypes produced pufM transcripts in situ. Enrichment cultures for purple bacteria yielded two morphotypes, each with a pufM signature identical to signatures detected by environmental screening. The isolates also contained gas vesicles, buoyancy structures previously unknown in purple nonsulfur bacteria, that may be necessary for these organisms to position themselves at specific depths within the nearly freezing water column. PMID- 12902287 TI - Colonization of Arabidopsis thaliana with Salmonella enterica and enterohemorrhagic Escherichia coli O157:H7 and competition by Enterobacter asburiae. AB - Enteric pathogens, such as Salmonella enterica and Escherichia coli O157:H7, have been shown to contaminate fresh produce. Under appropriate conditions, these bacteria will grow on and invade the plant tissue. We have developed Arabidopsis thaliana (thale cress) as a model system with the intention of studying plant responses to human pathogens. Under sterile conditions and at 100% humidity, S. enterica serovar Newport and E. coli O157:H7 grew to 10(9) CFU g(-1) on A. thaliana roots and to 2 x 10(7) CFU g(-1) on shoots. Furthermore, root inoculation led to contamination of the entire plant, indicating that the pathogens are capable of moving on or within the plant in the absence of competition. Inoculation with green fluorescent protein-labeled S. enterica and E. coli O157:H7 showed invasion of the roots at lateral root junctions. Movement was eliminated and invasion decreased when nonmotile mutants of S. enterica were used. Survival of S. enterica serovar Newport and E. coli O157:H7 on soil-grown plants declined as the plants matured, but both pathogens were detectable for at least 21 days. Survival of the pathogen was reduced in unautoclaved soil and amended soil, suggesting competition from indigenous epiphytes from the soil. Enterobacter asburiae was isolated from soil-grown A. thaliana and shown to be effective at suppressing epiphytic growth of both pathogens under gnotobiotic conditions. Seed and chaff harvested from contaminated plants were occasionally contaminated. The rate of recovery of S. enterica and E. coli O157:H7 from seed varied from undetectable to 19% of the seed pools tested, depending on the method of inoculation. Seed contamination by these pathogens was undetectable in the presence of the competitor, Enterobacter asburiae. Sampling of 74 pools of chaff indicated a strong correlation between contamination of the chaff and seed (P = 0.025). This suggested that contamination of the seed occurred directly from contaminated chaff or by invasion of the flower or silique. However, contaminated seeds were not sanitized by extensive washing and chlorine treatment, indicating that some of the bacteria reside in a protected niche on the seed surface or under the seed coat. PMID- 12902289 TI - Is the in situ accessibility of the 16S rRNA of Escherichia coli for Cy3-labeled oligonucleotide probes predicted by a three-dimensional structure model of the 30S ribosomal subunit? AB - Systematic studies on the hybridization of fluorescently labeled, rRNA-targeted oligonucleotides have shown strong variations in in situ accessibility. Reliable predictions of target site accessibility would contribute to more-rational design of probes for the identification of individual microbial cells in their natural environments. During the past 3 years, numerous studies of the higher-order structure of the ribosome have advanced our understanding of its spatial conformation. These studies range from the identification of rRNA-rRNA interactions based on covariation analyses to physical imaging of the ribosome for the identification of protein-rRNA interactions. Here we reevaluate our Escherichia coli 16S rRNA in situ accessibility data with regard to a tertiary structure model of the small subunit of the ribosome. We localized target sequences of 176 oligonucleotides on a 3.0-A-resolution three-dimensional (3D) model of the 30S ribosomal subunit. Little correlation was found between probe hybridization efficiency and the proximity of the probe target region to the surface of the 30S ribosomal subunit model. We attribute this to the fact that fluorescence in situ hybridization is performed on fixed cells containing denatured ribosomes, whereas 3D models of the ribosome are based on its native conformation. The effects of different fixation and hybridization protocols on the fluorescence signals conferred by a set of 10 representative probes were tested. The presence or absence of the strongly denaturing detergent sodium dodecyl sulfate had a much more pronounced effect than a change of fixative from paraformaldehyde to ethanol. PMID- 12902288 TI - Metagenomic profiling: microarray analysis of an environmental genomic library. AB - Genomic libraries derived from environmental DNA (metagenomic libraries) are useful for characterizing uncultured microorganisms. However, conventional library-screening techniques permit characterization of relatively few environmental clones. Here we describe a novel approach for characterization of a metagenomic library by hybridizing the library with DNA from a set of groundwater isolates, reference strains, and communities. A cosmid library derived from a microcosm of groundwater microorganisms was used to construct a microarray (COSMO) containing approximately 1-kb PCR products amplified from the inserts of 672 cosmids plus a set of 16S ribosomal DNA controls. COSMO was hybridized with Cy5-labeled genomic DNA from each bacterial strain, and the results were compared with the results for a common Cy3-labeled reference DNA sample consisting of a composite of genomic DNA from multiple species. The accuracy of the results was confirmed by the preferential hybridization of each strain to its corresponding rDNA probe. Cosmid clones were identified that hybridized specifically to each of 10 microcosm isolates, and other clones produced positive results with multiple related species, which is indicative of conserved genes. Many clones did not hybridize to any microcosm isolate; however, some of these clones hybridized to community genomic DNA, suggesting that they were derived from microbes that we failed to isolate in pure culture. Based on identification of genes by end sequencing of 17 such clones, DNA could be assigned to functions that have potential ecological importance, including hydrogen oxidation, nitrate reduction, and transposition. Metagenomic profiling offers an effective approach for rapidly characterizing many clones and identifying the clones corresponding to unidentified species of microorganisms. PMID- 12902290 TI - Use of 16S-23S rRNA intergenic spacer region PCR and repetitive extragenic palindromic PCR analyses of Escherichia coli isolates to identify nonpoint fecal sources. AB - Despite efforts to minimize fecal input into waterways, this kind of pollution continues to be a problem due to an inability to reliably identify nonpoint sources. Our objective was to find candidate source-specific Escherichia coli fingerprints as potential genotypic markers for raw sewage, horses, dogs, gulls, and cows. We evaluated 16S-23S rRNA intergenic spacer region (ISR)-PCR and repetitive extragenic palindromic (rep)-PCR analyses of E. coli isolates as tools to identify nonpoint fecal sources. The BOXA1R primer was used for rep-PCR analysis. A total of 267 E. coli isolates from different fecal sources were typed with both techniques. E. coli was found to be highly diverse. Only two candidate source-specific E. coli fingerprints, one for cow and one for raw sewage, were identified out of 87 ISR fingerprints. Similarly, there was only one candidate source-specific E. coli fingerprint for horse out of 59 BOX fingerprints. Jackknife analysis resulted in an average rate of correct classification (ARCC) of 83% for BOX-PCR analysis and 67% for ISR-PCR analysis for the five source categories of this study. When nonhuman sources were pooled so that each isolate was classified as animal or human derived (raw sewage), ARCCs of 82% for BOX-PCR analysis and 72% for ISR-PCR analysis were obtained. Critical factors affecting the utility of these methods, namely sample size and fingerprint stability, were also assessed. Chao1 estimation showed that generally 32 isolates per fecal source individual were sufficient to characterize the richness of the E. coli population of that source. The results of a fingerprint stability experiment indicated that BOX and ISR fingerprints were stable in natural waters at 4, 12, and 28 degrees C for 150 days. In conclusion, 16S-23S rRNA ISR-PCR and rep-PCR analyses of E. coli isolates have the potential to identify nonpoint fecal sources. A fairly small number of isolates was needed to find candidate source specific E. coli fingerprints that were stable under the simulated environmental conditions. PMID- 12902292 TI - Identification and characterization of two subpopulations of Encephalitozoon intestinalis. AB - Microsporidia are obligate intracellular protozoa that have been shown to be pathogenic to most living creatures. The development of in vitro cell culture propagation methods has provided researchers with large numbers of spores and facilitated the study of these organisms. Here, we describe heterogeneity within cell culture-propagated Encephalitozoon intestinalis suspensions. Flow cytometer histograms depicting the log side scatter and forward-angle light scatter of spores from nine suspensions produced over 12 months consistently showed two populations differing in size. The suspensions were composed primarily of the smaller-spore subpopulation (76.4% +/- 5.1%). The presence of two subpopulations was confirmed by microscopic examination and image analysis (P < 0.001). Small subpopulation spores were noninfectious in rabbit kidney (RK13) cell culture infectivity assays, while the large spores were infectious when inocula included > or = 25 spores. The small spores stained brilliantly with fluorescein isothiocyanate-conjugated monoclonal antibody against Encephalitozoon genus spore wall antigen, while the large spores stained poorly. There was no difference in staining intensities using commercial (MicroSporFA) and experimental polyclonal antibodies. Vital-dye (DAPI [4',6'-diamidino-2-phenylindole], propidium iodide, or SYTOX Green) staining showed the spores of the small subpopulation to be permeable to all vital dyes tested, while spores of the large subpopulation were not permeable in the absence of ethanol pretreatment. PCR using primers directed to the 16S rRNA or beta-tubulin genes and subsequent sequence analysis confirmed both subpopulations as E. intestinalis. Our data suggest that existing cell culture propagation methods produce two types of spores differing in infectivity, and the presence of these noninfective spores in purified spore suspensions should be considered when designing disinfection and drug treatment studies. PMID- 12902293 TI - Identification and application of plasmids suitable for transfer of foreign DNA to members of the genus Gordonia. AB - Gene transfer systems for Gordonia polyisoprenivorans strains VH2 and Y2K based on electroporation and conjugation, respectively, were established. Several parameters were optimized, resulting in transformation efficiencies of >4 x 10(5) CFU/ micro g of plasmid DNA. In contrast to most previously described electroporation protocols, the highest efficiencies were obtained by applying a heat shock after the intrinsic electroporation. Under these conditions, transfer and autonomous replication of plasmid pNC9503 was also demonstrated to proceed in G. alkanivorans DSM44187, G. nitida DSM44499(T), G. rubropertincta DSM43197(T), G. rubropertincta DSM46038, and G. terrae DSM43249(T). Conjugational plasmid DNA transfer to G. polyisoprenivorans resulted in transfer frequencies of up to 5 x 10(-6) of the recipient cells. Recombinant strains capable of polyhydroxyalkanoate synthesis from alkanes were constructed. PMID- 12902291 TI - Overexpression of groESL in Clostridium acetobutylicum results in increased solvent production and tolerance, prolonged metabolism, and changes in the cell's transcriptional program. AB - DNA array and Western analyses were used to examine the effects of groESL overexpression and host-plasmid interactions on solvent production in Clostridium acetobutylicum ATCC 824. Strain 824(pGROE1) was created to overexpress the groESL operon genes from a clostridial thiolase promoter. The growth of 824(pGROE1) was inhibited up to 85% less by a butanol challenge than that of the control strain, 824(pSOS95del). Overexpression of groESL resulted in increased final solvent titers 40% and 33% higher than those of the wild type and plasmid control strains, respectively. Active metabolism lasted two and one half times longer in 824(pGROE1) than in the wild type. Transcriptional analysis of 824(pGROE1) revealed increased expression of motility and chemotaxis genes and a decrease in the expression of the other major stress response genes. Decreased expression of the dnaKJ operon upon overexpression of groESL suggests that groESL functions as a modulator of the CIRCE regulon, which is shown here to include the hsp90 gene. Analysis of the plasmid control strain 824(pSOS95del) revealed complex host plasmid interactions relative to the wild-type strain, resulting in prolonged biphasic growth and metabolism. Decreased expression of four DNA gyrases resulted in differential expression of many key primary metabolism genes. The ftsA and ftsZ genes were expressed at higher levels in 824(pSOS95del), revealing an altered cell division and sporulation pattern. Both transcriptional and Western analyses revealed elevated stress protein expression in the plasmid-carrying strain. PMID- 12902294 TI - Description of a "phoenix" phenomenon in the growth of Campylobacter jejuni at temperatures close to the minimum for growth. AB - When Campylobacter jejuni cultures that had been grown in broth at 39 degrees C were subcultured into fresh medium at 30 degrees C, there was a transient period of growth followed by a decline in viable-cell numbers before growth resumed once more. We propose that this complex behavior is the net effect of the growth of inoculum cells followed by a loss of viability due to oxidative stress and the subsequent emergence of a spontaneously arising mutant population that takes over the culture. PMID- 12902295 TI - Analysis of the argK-tox gene cluster in nontoxigenic strains of Pseudomonas syringae pv. phaseolicola. AB - The analysis of 46 isolates obtained directly from different and distant common bean fields from the northwestern part of Spain revealed that they do not produce phaseolotoxin. The isolates were classified as race 5, and their analysis revealed that they do not carry the argK-tox gene cluster involved in the biosynthesis of the phaseolotoxin. PMID- 12902296 TI - Infectious Cryptosporidium parvum oocysts in final reclaimed effluent. AB - Water samples collected throughout several reclamation facilities were analyzed for the presence of infectious Cryptosporidium parvum by the focus detection method-most-probable-number cell culture technique. Results revealed the presence of infectious C. parvum oocysts in 40% of the final disinfected effluent samples. Sampled effluent contained on average seven infectious oocysts per 100 liters. Thus, reclaimed water is not pathogen free but contains infectious C. parvum. PMID- 12902297 TI - Development of a sensitive gene expression reporter system and an inducible promoter-repressor system for Clostridium acetobutylicum. AB - A sensitive gene expression reporter system was developed for Clostridium acetobutylicum ATCC 824 by using a customized gusA expression cassette. In discontinuous cultures, time course profiles of beta-glucuronidase specific activity reflected adequately in vivo dynamic up- and down-regulation of acidogenesis- and/or solventogenesis-associated promoter expression in C. acetobutylicum. Furthermore, a new inducible gene expression system was developed in C. acetobutylicum, based on the Staphylococcus xylosus xylose operon promoter repressor regulatory system. PMID- 12902298 TI - The Ti plasmid of Agrobacterium tumefaciens harbors an attM-paralogous gene, aiiB, also encoding N-Acyl homoserine lactonase activity. AB - The Agrobacterium tumefaciens C58 genome contains three putative N-acyl homoserine lactone (acyl-HSL) hydrolases, which are closely related to the lactonase AiiA of Bacillus. When expressed in Escherichia coli, two of the putative acyl-HSL hydrolases, AttM and AiiB, conferred the ability to degrade acyl-HSLs on the host. In Erwinia strain 6276, the lactonases reduced the endogenous acyl-HSL level and the bacterial virulence in planta. PMID- 12902299 TI - Internal and external mycoflora of the American dog tick, Dermacentor variabilis (Acari: Ixodidae), and its ecological implications. AB - Scopulariopsis brevicaulis, the anamorph of Microascus brevicaulis (Microascaceae, Ascomycota), has been identified in the body contents of the tick Dermacentor variabilis. After topical application of the fungal inoculum, tick mortality was marked. This is the first account describing the internal mycoflora of D. variabilis with a novel technique used to recover potential biological control agents. PMID- 12902300 TI - Microbial isotopic fractionation of perchlorate chlorine. AB - Perchlorate contamination can be microbially respired to innocuous chloride and thus can be treated effectively. However, monitoring a bioremediative strategy is often difficult due to the complexities of environmental samples. Here we demonstrate that microbial respiration of perchlorate results in a significant fractionation ( approximately -15 per thousand ) of the chlorine stable isotope composition of perchlorate. This can be used to quantify the extent of biotic degradation and to separate biotic from abiotic attenuation of this contaminant. PMID- 12902302 TI - Boundaries for biofilm formation: humidity and temperature. AB - Environmental conditions which define boundaries for biofilm production could provide useful ecological information for biofilm models. A practical use of defined conditions could be applied to the high-level nuclear waste repository at Yucca Mountain. Data for temperature and humidity conditions indicate that decreases in relative humidity or increased temperature severely affect biofilm formation on three candidate canister metals. PMID- 12902301 TI - Characterization of Anaplasma marginale isolated from North American bison. AB - Anaplasma marginale (Rickettsiales: Anaplasmataceae), a tick-borne pathogen of cattle, is endemic in tropical and subtropical regions of the world. Although serologic tests have identified American bison, Bison bison, as being infected with A. marginale, the present study was undertaken to confirm A. marginale infection and to characterize isolates obtained from naturally infected bison in the United States and Canada. Major surface protein (MSP1a and MSP4) sequences of bison isolates were characterized in comparison with New World cattle isolates. Blood from one U.S. bison was inoculated into a susceptible, splenectomized calf, which developed acute anaplasmosis, demonstrating infectivity of this A. marginale bison isolate for cattle. The results of this study showed that these A. marginale isolates obtained from bison were similar to ones from naturally infected cattle. PMID- 12902303 TI - Effects of essential oils on ruminal microorganisms and their protein metabolism. AB - A commercial blend of essential oil (EO) compounds was added to a grass, maize silage, and concentrate diet fed to dairy cattle in order to determine their influence on protein metabolism by ruminal microorganisms. EO inhibited (P < 0.05) the rate of deamination of amino acids. Pure-culture studies indicated that the species most sensitive to EO were ammonia-hyperproducing bacteria and anaerobic fungi. PMID- 12902304 TI - Chlorination and cleavage of lignin structures by fungal chloroperoxidases. AB - Two fungal chloroperoxidases (CPOs), the heme enzyme from Caldariomyces fumago and the vanadium enzyme from Curvularia inaequalis, chlorinated 1-(4-ethoxy-3 methoxyphenyl)-2-(2-methoxyphenoxy)-1,3-dihydroxypropane, a dimeric model compound that represents the major nonphenolic structure in lignin. Both enzymes also cleaved this dimer to give 1-chloro-4-ethoxy-3-methoxybenzene and 1,2 dichloro-4-ethoxy-5-methoxybenzene, and they depolymerized a synthetic guaiacyl lignin. Since fungal CPOs occur in soils and the fungi that produce them are common inhabitants of plant debris, CPOs may have roles in the natural production of high-molecular-weight chloroaromatics and in lignin breakdown. PMID- 12902305 TI - Structures of the mating-type loci of Cordyceps takaomontana. AB - Nucleotide sequences of the mating-type loci MAT1-1 and MAT1-2 of Cordyceps takaomontana were determined, which is the first such report for the clavicipitaceous fungi. MAT1-1 contains two mating-type genes, MAT1-1-1 and MAT1 1-2, but MAT1-1-3 could not be found. On the other hand, MAT1-2 has MAT1-2-1. A pseudogene of MAT1-1-1 is located next to MAT1-2. PMID- 12902306 TI - Detection of bacteria carrying the stx2 gene by in situ loop-mediated isothermal amplification. AB - A new in situ DNA amplification technique for microscopic detection of bacteria carrying a specific gene is described. Loop-mediated isothermal amplification (LAMP) was used to detect stxA(2) in Escherichia coli O157:H7 cells. The mild permeabilization conditions and low isothermal temperature used in the in situ LAMP method caused less cell damage than in situ PCR. It allowed use of fluorescent antibody labeling in the bacterial mixture after the DNA amplification for identification of E. coli O157:H7 cells with an stxA(2) gene. Higher-contrast images were obtained with this method than with in situ PCR. PMID- 12902307 TI - Increased exopolysaccharide production in Lactococcus lactis due to increased levels of expression of the NIZO B40 eps gene cluster. AB - Exopolysaccharides (EPS) play an important role in the rheology and texture of fermented food products. This is the first report demonstrating that homologous overexpression of a complete eps gene cluster in Lactococcus lactis leads to increased EPS production levels. A ninefold-elevated EPS plasmid copy number led to an almost threefold increase in the eps expression level, resulting in an almost fourfold increase in the NIZO B40 EPS production level. It was previously reported that increased EPS precursor levels did not influence NIZO B40 EPS production levels. However, the present results indicate that the maximal NIZO B40 EPS production level is limited by the activity level of the expression products of the eps gene cluster rather than by the level of EPS precursors. PMID- 12902308 TI - Reduction of experimental Salmonella and Campylobacter contamination of chicken skin by application of lytic bacteriophages. AB - Lytic bacteriophages, applied to chicken skin that had been experimentally contaminated with Salmonella enterica serovar Enteritidis or Campylobacter jejuni at a multiplicity of infection (MOI) of 1, increased in titer and reduced the pathogen numbers by less than 1 log(10) unit. Phages applied at a MOI of 100 to 1,000 rapidly reduced the recoverable bacterial numbers by up to 2 log(10) units over 48 h. When the level of Salmonella contamination was low (< log(10) 2 per unit area of skin) and the MOI was 10(5), no organisms were recovered. By increasing the number of phage particles applied (i.e., MOI of 10(7)), it was also possible to eliminate other Salmonella strains that showed high levels of resistance because of restriction but to which the phages were able to attach. PMID- 12902309 TI - Depletion of ventromedullary NK-1 receptor-immunoreactive neurons in multiple system atrophy. AB - We sought to determine whether there are neurokinin-1 receptor-like immunoreactive (NK-1R-LI) neurons in human ventrolateral medulla and whether these neurons are more severely involved in multiple system atrophy (MSA) than in Parkinson's disease. Brains were obtained at autopsy from six control subjects, six subjects with clinical diagnosis of MSA and four with Parkinson's disease, both confirmed neuropathologically. Serial 50 microm cryostat sections were obtained throughout the medulla, and every eighth section was processed for NK-1R LI neurons. Some sections were processed simultaneously for tyrosine hydroxylase or choline acetyltransferase. Abundant NK-1R-LI neurons were identified in the ventrolateral medulla. These neurons were distinct from local cholinergic or catecholaminergic neurons. There was a severe depletion of these NK-1R-LI neurons in all MSA cases compared with controls (6 +/- 1 cells/section versus 49 +/- 2 cells/section in controls). Although there was also a reduction in Parkinson's disease (20 +/- 2 cells/section), this was significantly less severe than in MSA. Our findings suggest that the human ventrolateral medulla contains NK-1R-LI neurons, and the more severe depletion in MSA than in Parkinson's disease may explain the higher incidence of respiratory and cardiovascular abnormalities in the former condition. PMID- 12902310 TI - Neural correlates of laughter and humour. AB - Although laughter and humour have been constituents of humanity for thousands if not millions of years, their systematic study has begun only recently. Investigations into their neurological correlates remain fragmentary and the following review is a first attempt to collate and evaluate these studies, most of which have been published over the last two decades. By employing the classical methods of neurology, brain regions associated with symptomatic (pathological) laughter have been determined and catalogued under other diagnostic signs and symptoms of such conditions as epilepsy, strokes and circumspect brain lesions. These observations have been complemented by newer studies using modern non-invasive imaging methods. To summarize the results of many studies, the expression of laughter seems to depend on two partially independent neuronal pathways. The first of these, an 'involuntary' or 'emotionally driven' system, involves the amygdala, thalamic/hypo- and subthalamic areas and the dorsal/tegmental brainstem. The second, 'voluntary' system originates in the premotor/frontal opercular areas and leads through the motor cortex and pyramidal tract to the ventral brainstem. These systems and the laughter response appear to be coordinated by a laughter-coordinating centre in the dorsal upper pons. Analyses of the cerebral correlates of humour have been impeded by a lack of consensus among psychologists on exactly what humour is, and of what essential components it consists. Within the past two decades, however, sufficient agreement has been reached that theory-based hypotheses could be formulated and tested with various non-invasive methods. For the perception of humour (and depending on the type of humour involved, its mode of transmission, etc.) the right frontal cortex, the medial ventral prefrontal cortex, the right and left posterior (middle and inferior) temporal regions and possibly the cerebellum seem to be involved to varying degrees. An attempt has been made to be as thorough as possible in documenting the foundations upon which these burgeoning areas of research have been based up to the present time. PMID- 12902312 TI - Idiom comprehension in Alzheimer's disease: the role of the central executive. AB - Idiom comprehension of 15 patients with mild probable Alzheimer's disease was examined by means of a sentence-to-picture matching task. Patients had to choose between two pictures, one representing the figurative and the other the literal interpretation. They were also submitted to a literal sentence comprehension test and to a pencil-and-paper dual task. Whereas literal comprehension was normal in seven subjects and mildly impaired in the others, idiom comprehension was very poor in all of them and correlated with the performance on the dual task. When the idiom test was repeated using an unrelated situation as an alternative to the picture representing the figurative meaning, performance significantly improved. It was hypothesized that the response in the sentence-to-picture matching task in the case of idioms requires sentence processing followed by the suppression of the literal interpretation. Alzheimer's disease patients proved to be unable to inhibit the literal meaning, although they had not lost the idiomatic meaning. In a second experiment, 15 Alzheimer's disease patients with a comparable level of cognitive impairment were submitted to the same idiom comprehension test, and to a test of verbal explanation of the idioms. The results showed significantly better performance in the oral task than in the sentence-to-picture matching task. In oral explanation, however, Alzheimer's disease patients also produced some literal interpretation whenever this represented a possible situation in the real world. We suggest that, during idiom interpretation, the literal meaning needs to be suppressed in order to activate the figurative meaning, and we stress the fact that both linguistic and extralinguistic factors must be taken into account to explain idiom interpretation. PMID- 12902311 TI - Progressive non-fluent aphasia is associated with hypometabolism centred on the left anterior insula. AB - Progressive non-fluent aphasia (PNFA) is a syndrome in which patients lose the ability to communicate fluently in the context of relative preservation of single word comprehension and non-linguistic cognitive abilities. Neuroimaging in case studies with PNFA has failed to identify a consistent neural substrate for the language disorder. In this study of a group of patients (n=10) whose presenting complaint was progressive dysfluency, resting cerebral metabolism was measured using [18F]fluorodeoxyglucose-PET and analysed with the technique of statistical parametric mapping (SPM). Regional atrophy was assessed with voxel-based morphometry (VBM). Seven patients had a 'pure' PNFA syndrome, while the remaining three had additional features of a more pervasive dementia. Compared with controls, the patients showed hypometabolism in several regions that, most notably, included the left anterior insula/frontal opercular region. The VBM analysis revealed only one small area of atrophy in the left peri-Sylvian region. Analysis of the pure PNFA cases (n=7) relative to controls yielded qualitatively similar results to those of the whole group, suggesting that these cases were also at risk of a more generalized dementia, a finding borne out in subsequent follow-up of two cases to date. The PNFA group was then compared with a group with Alzheimer's disease (n=10) whose clinical profile did not include non-fluent aphasic features. In this analysis, the only persisting hypometabolic region was that centred over the left anterior insula. VBM did not identify any regional differences in atrophy between PNFA and Alzheimer's disease. In the light of current theories of fluent language production, the findings offer anatomical evidence that the breakdown in fluency is due to a motor articulatory planning deficit (speech apraxia) combined with a variable degree of agrammatism. PMID- 12902313 TI - Magnetic resonance spectroscopy and imaging of the thalamus in idiopathic generalized epilepsy. AB - Experimental work in animal models of generalized epilepsy and clinical data in humans with idiopathic generalized epilepsy (IGE) indicate that the thalamo cortical circuitry is involved in the generation of epileptic activity. The purpose of this study was to evaluate in vivo the chemical and structural integrity of the thalamus in patients with IGE. Thalamic proton magnetic resonance spectroscopic imaging (1H-MRSI), measuring N-acetylaspartate (NAA), choline-containing compounds and creatine (Cr) was performed in 20 IGE patients and in a group of age-matched healthy subjects. Additionally, 1H-MRSI measurements were taken in the insular cortex, the posterior temporal lobe white matter and the splenium of the corpus callosum. MRI volumetric analysis of the thalamus was performed in all patients. At the time of the examination, seizures were well controlled in 10 IGE patients and poorly controlled in nine. One patient was newly diagnosed and had the MRI and MRSI examination prior to starting the antiepileptic medication. In IGE patients, 1H-MRSI showed a reduction of mean thalamic NAA/Cr compared with normal controls; no difference was found in NAA/Cr in the other examined areas. There was no difference in NAA/Cr between patients whose seizures were well controlled and those in whom seizures were not controlled. There was no correlation between thalamic NAA/Cr and mean number of spike and wave complexes. We found a significant negative correlation between thalamic NAA/Cr and duration of epilepsy. The mean thalamic volume in patients with IGE was not different from normal controls. These results show evidence of progressive thalamic neuronal dysfunction in patients with IGE supporting the notion of abnormal thalamo-cortical circuitry as a substrate of seizure generation in this form of epilepsy. The thalamic dysfunction may occur regardless of amount of spike and wave activity. PMID- 12902314 TI - Primary progressive multiple sclerosis: a 5-year clinical and MR study. AB - Longitudinal imaging studies of primary progressive multiple sclerosis (PPMS) have shown significant changes in MR measures over 1 to 2 years. Correlation with clinical change over the same period has not been evident; we investigated the possibility that this is because the period of observation was insufficient for these associations to become apparent. Forty-one patients with PPMS were followed prospectively for 5 years. Patients had clinical [Expanded Disability Status Scale (EDSS) and Multiple Sclerosis Functional Composite Measure (MSFC)] and MRI assessment (brain and spinal cord) at baseline, 1, 2 and 5 years. At 5 years, significant deterioration was seen in all clinical and MRI measures (P<0.01, P<0.001 respectively). Associations were seen between increase in EDSS score and decrease in cord area (r=0.31, P<0.05) and between increase in MSFC and both rate of ventricular enlargement (r=0.31, P<0.05) and increase in T2 load (r=0.31, P<0.05). The rates of change of MR measures were not associated with age or disease duration and were more consistent within than between patients. Longer duration of follow-up demonstrates modest associations between change in clinical and MR measures and provides new insights into the pattern of change within and between individuals with PPMS. PMID- 12902315 TI - Thoughts on the pulmonary blood-gas barrier. AB - The pulmonary blood-gas barrier is an extraordinary structure because of its extreme thinness, immense strength, and enormous area. The essential components of the barrier were determined early in evolution and have been highly conserved. For example, the barriers of the African, Australian, and South American lungfish that date from as much as 400 million years ago have essentially the same structure as in the modern mammal or bird. In the evolution of vertebrates from bony fishes through amphibia, reptiles, and ultimately mammals and birds, changes in the pulmonary circulation occurred to limit the stresses in the blood-gas barrier. Only in mammals and birds is there a complete separation of the pulmonary and systemic circulations, which is essential to protect the extremely thin barrier from the necessary high-vascular pressures. To provide the blood-gas barrier with its required strength, evolution has exploited the high ultimate tensile strength of type IV collagen in basement membrane. Nevertheless, stress failure of the barrier occurs under physiological conditions in galloping Thoroughbred racehorses and also apparently in elite human athletes at maximal exercise. The human blood-gas barrier maintains its integrity during all but the most extreme physiological conditions. However, many pathological conditions cause stress failure. The structure of the blood-gas barrier is apparently continually regulated in response to wall stress, and this regulation is essential to maintain the extreme thinness but adequate strength. The mechanisms of this regulation remain to be elucidated and constitute one of the fundamental problems in lung biology. PMID- 12902316 TI - Transforming growth factor-beta induces fibrosis in immune cell-depleted lungs. PMID- 12902317 TI - Surfactant protein B: unambiguously necessary for adult pulmonary function. PMID- 12902318 TI - Surface activity of a synthetic lung surfactant containing a phospholipase resistant phosphonolipid analog of dipalmitoyl phosphatidylcholine. AB - Surface activity and sensitivity to inhibition from phospholipase A2 (PLA2), lysophosphatidylcholine (LPC), and serum albumin were studied for a synthetic C16:0 diether phosphonolipid (DEPN-8) combined with 1.5% by weight of mixed hydrophobic surfactant proteins (SP)-B/C purified from calf lung surfactant extract (CLSE). Pure DEPN-8 had better adsorption and film respreading than the major lung surfactant phospholipid dipalmitoyl phosphatidylcholine and reached minimum surface tensions <1 mN/m under dynamic compression on the Wilhelmy balance and on a pulsating bubble surfactometer (37 degrees C, 20 cycles/min, 50% area compression). DEPN-8 + 1.5% SP-B/C exhibited even greater adsorption and had overall dynamic surface tension lowering equal to CLSE on the bubble. In addition, films of DEPN-8 + 1.5% SP-B/C on the Wilhelmy balance had better respreading than CLSE after seven (but not two) cycles of compression-expansion at 23 degrees C. DEPN-8 is structurally resistant to degradation by PLA2, and DEPN-8 + 1.5% SP-B/C maintained high adsorption and dynamic surface activity in the presence of this enzyme. Incubation of CLSE with PLA2 led to chemical degradation, generation of LPC, and reduced surface activity. DEPN-8 + 1.5% SP B/C was also more resistant than CLSE to direct biophysical inhibition by LPC, and the two were similar in their sensitivity to biophysical inhibition by serum albumin. These findings indicate that synthetic surfactants containing DEPN-8 combined with surfactant proteins or related synthetic peptides have potential utility for treating surfactant dysfunction in inflammatory lung injury. PMID- 12902319 TI - Isocaloric maternal low-protein diet alters IGF-I, IGFBPs, and hepatocyte proliferation in the fetal rat. AB - We investigated the effect of an isocaloric maternal low-protein diet during pregnancy in rats on the proliferative capacity of cultured fetal hepatocytes. The potential roles of these changes on the IGF-IGF-binding protein (IGFBP) axis, and the role of insulin and glucocorticoids in liver growth retardation, were also evaluated. Pregnant Wistar rats were fed a control (C) diet (20% protein) or a low-protein (LP) diet (8%) throughout gestation. In primary culture, the DNA synthesis of hepatocytes derived from LP fetuses was decreased by approximately 30% compared with control hepatocytes (P < 0.05). In parallel, in vivo moderate protein restriction in the dam reduced the fetal liver weight and IGF-I level in fetal plasma (P < 0.01) and augmented the abundance of 29- to 32-kDa IGFBPs in fetal plasma (P < 0.01) and fetal liver (P < 0.01). By contrast, the abundance of IGF-II mRNA in liver of LP fetuses was unaffected by the LP diet. In vitro, the LP-derived hepatocytes produced less IGF-I (P < 0.01) and more 29- to 32-kDa IGFBPs (P < 0.01) than hepatocytes derived from control fetuses. These alterations still appeared after 3-4 days of culture, indicating some persistence in programming. Dexamethasone treatment of control-derived hepatocytes decreased cell proliferation (54 +/- 2.3%, P < 0.01) and stimulated 29- to 32-kDa IGFBPs, whereas insulin promoted fetal hepatocyte growth (127 +/- 5.5%, P < 0.01) and inhibited 29- to 32-kDa IGFBPs. These results show that liver growth and cell proliferation in association with IGF-I and IGFBP levels are affected in utero by fetal undernutrition. It also suggests that glucocorticoids and insulin may modulate these effects. PMID- 12902320 TI - Characterization of an organic anion transport system in a placental cell line. AB - Transporters within the placenta play a crucial role in the distribution of nutrients and xenobiotics across the maternal-fetal interface. An organic anion transport system was identified on the apical membrane of the rat placenta cell line HRP-1, a model for the placenta barrier. The apical uptake of 3H-labeled organic anion estrone sulfate in HRP-1 cells was saturable (Km = 4.67 microM), temperature and Na+ dependent, Li+ tolerant, and pH sensitive. The substrate specificity of the transport system includes various steroid sulfates, such as beta-estradiol 3,17-disulfate, 17 beta-estradiol 3-sulfate, and dehydroepiandrosterone 3-sulfate (DHEAS) but does not include taurocholate, p aminohippuric acid (PAH), and tetraethylammonium. Preincubation of HRP-1 cells with 8-bromo-cAMP (a cAMP analog) and forskolin (an adenylyl cyclase activator) acutely stimulated the apical transport activity. This stimulation was further enhanced in the presence of IBMX (a phosphodiesterase inhibitor). Together these data show that the apical membrane of HRP-1 cells expresses an organic anion transport system that is regulated by cellular cAMP levels. This transport system appears to be different from the known taurocholate-transporting organic anion transporting polypeptides and PAH-transporting organic anion transporters, both of which also mediate the transport of estrone sulfate and DHEAS. PMID- 12902321 TI - Effects of depletion exercise and light training on muscle glycogen supercompensation in men. AB - Supercompensated muscle glycogen can be achieved by using several carbohydrate (CHO)-loading protocols. This study compared the effectiveness of two "modified" CHO-loading protocols. Additionally, we determined the effect of light cycle training on muscle glycogen. Subjects completed a depletion (D, n = 15) or nondepletion (ND, n = 10) CHO-loading protocol. After a 2-day adaptation period in a metabolic ward, the D group performed a 120-min cycle exercise at 65% peak oxygen uptake (VO2 peak) followed by 1-min sprints at 120% VO2 peak to exhaustion. The ND group performed only 20-min cycle exercise at 65% VO2 peak. For the next 6 days, both groups ate the same high-CHO diets and performed 20-min daily cycle exercise at 65% VO2 peak followed by a CHO beverage (105 g of CHO). Muscle glycogen concentrations of the vastus lateralis were measured daily with 13C magnetic resonance spectroscopy. On the morning of day 5, muscle glycogen concentrations had increased 1.45 (D) and 1.24 (ND) times baseline (P < 0.001) but did not differ significantly between groups. However, on day 7, muscle glycogen of the D group was significantly greater (p < 0.01) than that of the ND group (130 +/- 7 vs. 104 +/- 5 mmol/l). Daily cycle exercise decreased muscle glycogen by 10 +/- 2 (D) and 14 +/- 5 mmol/l (ND), but muscle glycogen was equal to or greater than preexercise values 24 h later. In conclusion, a CHO-loading protocol that begins with a glycogen-depleting exercise results in significantly greater muscle glycogen that persists longer than a CHO-loading protocol using only an exercise taper. Daily exercise at 65% VO2 peak for 20 min can be performed throughout the CHO-loading protocol without negatively affecting muscle glycogen supercompensation. PMID- 12902322 TI - Differential transcriptional activation of select metabolic genes in response to variations in exercise intensity and duration. AB - Cellular adaptations to endurance training are influenced by the intensity and duration of exercise. To examine the impact of exercise intensity and duration on the acute transcriptional regulation of metabolic genes in red (RG) and white (WG) gastrocnemius muscle, rats completed either low-intensity [ approximately 50% maximal O2 uptake (VO2 max)] treadmill exercise (LIE) for 45 min, LIE for 180 min, or high-intensity ( approximately 75% VO2 max) exercise (HIE) for 45 min. LIE for 45 min activated (P < 0.05) transcription of the pyruvate dehydrogenase kinase-4 (PDK4), uncoupling protein-3 (UCP3), heme oxygenase-1 (HO-1), and hexokinase II (HK II) genes in RG within 1 h after exercise. In WG, transcription of PDK4, UCP3, HKII, and lipoprotein lipase (LPL) was also induced, whereas transcription of the HO-1 gene did not change. In RG, extending LIE duration from 45 to 180 min elicited a similar activation of PDK4 and UCP3 ( approximately 15 fold) but a far greater increase in HO-1 (>30-fold) and HKII transcription (>25 fold). In WG, extending LIE for 180 min induced a much greater and prolonged (through 2- to 4-h recovery) activation of PDK4, UCP3 (both >200-fold), and HO-1 (>10-fold). HIE elicited a similar pattern of gene activation to LIE in both RG and WG, with the exception that HIE triggered >10-fold activation of HO-1 in WG. These data provide evidence that both the intensity and the duration of exercise affect the transcriptional regulation of metabolic genes in muscle in a fiber type-specific manner, possibly reflecting the relative stress imposed by the exercise bout. PMID- 12902323 TI - Roles of His291-alpha and His146-beta' in the reductive acylation reaction catalyzed by human branched-chain alpha-ketoacid dehydrogenase: refined phosphorylation loop structure in the active site. AB - We report here that alterations of either His291-alpha or His146-beta' in the active site of human branched-chain alpha-ketoacid dehydrogenase (E1b) impede both the decarboxylation and the reductive acylation reactions catalyzed by E1b as well as the binding of cofactor thiamin diphosphate (ThDP). In a refined human E1b active-site structure, His291-alpha, which aligns with His407 in Escherichia coli pyruvate dehydrogenase and His263 in yeast transketolase, is on a largely ordered phosphorylation loop. The imidazole ring of His291-alpha in E1b coordinates to the terminal phosphate oxygen atoms of bound ThDP. The N3 atom of wild-type His146-beta', which can be protonated, binds a water molecule and points toward the aminopyrimidine ring of ThDP. Remarkably, the H291A-alpha mutation results in a complete order-to-disorder transition of the loop region, which precludes the binding of the substrate lipoyl-bearing domain to E1b. The H146A-beta' mutation, on the other hand, does not alter the loop structure, but nullifies the reductive acylation activity of E1b. Our results suggest that: 1) His291-alpha plays a structural rather than a catalytic role in the binding of cofactor ThDP and the lipoyl-bearing domain to E1b, and 2) His146-beta' is an essential catalytic residue, probably functioning as a proton donor in the reductive acylation of lipoamide on the lipoyl-bearing domain. PMID- 12902324 TI - Myeloid differentiation factor 88-dependent post-transcriptional regulation of cyclooxygenase-2 expression by CpG DNA: tumor necrosis factor-alpha receptor associated factor 6, a diverging point in the Toll-like receptor 9-signaling. AB - The immune stimulatory unmethylated CpG motifs present in bacterial DNA (CpG DNA) induce expression of cyclooxygenase-2 (cox-2). The present study demonstrates that CpG DNA can up-regulate cox-2 expression by post-transcriptional mechanisms in RAW264.7 cells. To determine the CpG DNA-mediated signaling pathway that post transcriptionally regulates cox-2 expression, a cox-2 translational reporter (COX2-3'-UTR-luciferase) was generated by inserting sequences within the 3' untranslated region (UTR) of cox-2 to the 3' end of the luciferase gene under control of the SV40 promoter. CpG DNA-induced COX2-3'-UTR-luciferase activity was completely inhibited by an endosomal acidification inhibitor chloroquine, a Toll like receptor 9 antagonist inhibitory CpG DNA, or overexpression of a dominant negative (DN) form of MyD88. However, overexpression of DN-IRAK-1 or DN-TRAF6 resulted in substantial, but not complete, inhibition of the CpG DNA-induced COX2 3'-UTR-luciferase activity. Activation of all three MAPKs (ERK, p38, and JNK) was required for optimal COX2-3'-UTR-luciferase activity induced by CpG DNA. Overexpression of DN-TRAF6 suppressed CpG DNA-mediated activation of p38 and JNK, but not ERK, explaining the partial inhibitory effects of DN-TRAF6 on CpG DNA induced COX2-3'-UTR-luciferase activity. Co-expression of DN-TRAF6 and N17Ras completely inhibited CpG DNA-induced COX2-3'-UTR-luciferase activity, indicating the involvement of Ras in CpG DNA-mediated ERK and COX2-3'-UTR regulation. Collectively, our results suggest that MyD88 and MAPKs play a key regulatory role in CpG DNA-mediated cox-2 expression at the post-transcriptional level and that TRAF6 is a diverging point in the Toll-like receptor 9-signaling pathway for CpG DNA-mediated MAPK activation. PMID- 12902325 TI - Anti-migratory and anti-invasive effect of somatostatin in human neuroblastoma cells: involvement of Rac and MAP kinase activity. AB - Cell motility and invasion are crucial events for the spread of cancer and, consequently, the metastatic process. Platelet-derived growth factor (PDGF) is not only capable of stimulating the proliferation of SH-SY5Y human neuroblastoma cells, but also their migration and invasion through an extracellular matrix barrier. Experiments using wortmannin and PD98059, specific inhibitors of the phosphatidylinositol 3-kinase (PI3-K) and of the mitogen-activated protein kinases (ERK 1 and 2) signaling, respectively, show that the activation of both pathways is required for the PDGF-induced cell motility responses. We have previously shown that somatostatin inhibits cell division and ERK 1/2 and Ras activity in SH-SY5Y cells. We report here that it is also capable of potently and effectively inhibiting their PDGF-stimulated migration and invasion. The inhibitory effect of somatostatin is sensitive to pertussis toxin. Although somatostatin does not affect PI3-K, it inhibits ERK 1/2 and the small G-protein Rac activation and ruffle formation induced by PDGF. These results indicate that somatostatin can be considered an anti-migratory and anti-invasive agent that acts by inhibiting ERK 1/2 signaling and the PI3-K pathway via the inhibition of Rac in SHSY5Y cells. PMID- 12902326 TI - Catalytic and thermodynamic properties of tetrahydromethanopterin-dependent serine hydroxymethyltransferase from Methanococcus jannaschii. AB - The reaction catalyzed by serine hydroxymethyltransferase (SHMT), the transfer of Cbeta of serine to tetrahydropteroylglutamate, represents in Eucarya and Eubacteria a major source of one-carbon (C1) units for several essential biosynthetic processes. In many Archaea, C1 units are carried by modified pterin containing compounds, which, although structurally related to tetrahydropteroylglutamate, play a distinct functional role. Tetrahydromethanopterin, and a few variants of this compound, are the modified folates of methanogenic and sulfate-reducing Archaea. Little information on SHMT from Archaea is available, and the metabolic role of the enzyme in these organisms is not clear. This contribution reports on the purification and characterization of recombinant SHMT from the hyperthermophilic methanogen Methanococcus jannaschii. The enzyme was characterized with respect to its catalytic, spectroscopic, and thermodynamic properties. Tetrahydromethanopterin was found to be the preferential pteridine substrate. Tetrahydropteroylglutamate could also take part in the hydroxymethyltransferase reaction, although with a much lower efficiency. The catalytic features of the enzyme with substrate analogues and in the absence of a pteridine substrate were also very similar to those of SHMT isolated from Eucarya or Eubacteria. On the other hand, the M. jannaschii enzyme showed increased thermoactivity and resistance to denaturating agents with respect to the enzyme purified from mesophilic sources. The results reported suggest that the active site structure and the mechanism of SHMT are conserved in the enzyme from M. jannaschii, which appear to differ only in its ability to bind and use a modified folate as substrate and increased thermal stability. PMID- 12902327 TI - Protein-tyrosine phosphatase 1B mediates the effects of insulin on the actin cytoskeleton in immortalized fibroblasts. AB - Insulin regulates diverse cellular responses including actin reorganization. The mechanism by which insulin induces formation of lamellipodia in cultured cells is not known but is likely to involve activation of Src family protein-tyrosine kinases. Here we show that protein-tyrosine phosphatase 1B (PTPIB) activates Src, thereby initiating the activation of a Rac-dependent pathway leading to cytoskeletal remodeling. Conversely, expression of a proline to alanine (P309,310A) PTP1B mutant, which cannot activate Src, fails to activate Rho GTPases or cause changes in actin organization. Rat fibroblasts lacking PTP1B expression do not activate Src or Rac in response to insulin and cannot reorganize actin. These results show that PTP1B, best known as a negative regulator of the metabolic effects of insulin, is required for the effects of insulin on actin organization in immortalized fibroblasts. PMID- 12902328 TI - Mirk/dyrk1B is a Rho-induced kinase active in skeletal muscle differentiation. AB - The Rho family of small GTPases regulates numerous signaling pathways that control the organization of the cytoskeleton, transcription factor activity, and many aspects of the differentiation of skeletal myoblasts. We now demonstrate that the kinase Mirk (minibrain-related kinase)/dyrk1B is induced by members of the Rho-family in myoblasts and that Mirk is active in skeletal muscle differentiation. Mirk is an arginine-directed serine/threonine kinase which is expressed at elevated levels in skeletal muscle compared with other normal tissues. A Mirk promoter construct was activated when C2C12 myoblasts were switched from growth to differentiation medium and was also activated by the Rho family members RhoA, Cdc42, and to a lesser degree Rac1, but not by MyoD or Myf5. Mirk protein levels increased following transient expression of constitutively active Cdc42-QL, RhoA-QL, or Rac1-QL in C2C12 cells. High concentrations of serum mitogens down-regulated Mirk through activation of the Ras-MEK-Erk pathway. As a result, Mirk transcription was induced by the MEK1 inhibitor PD98059 and by the switch from growth to differentiation medium. Mirk was induced with similar kinetics to another Rho-induced differentiation gene, myogenin. Mirk protein levels increased 10-fold within 24-48 h after primary cultured muscle cells; C2C12 mouse myoblasts or L6 rat myoblasts were induced to differentiate. Thus Mirk was induced following the commitment stage of myogenesis. Stable overexpression of Mirk enabled myoblasts to fuse more rapidly when placed in differentiation medium. The function of Mirk in muscle differentiation was established by depletion of endogenous Mirk by small interfering RNA, which prevented myoblast fusion into myotubes and inhibited induction of markers of differentiation, including myogenin, fast twitch troponin T, and muscle myosin heavy chain. Other members of the dyrk/minibrain/HIPK family of kinases in lower organisms have been shown to regulate the transition from growth to differentiation, and Mirk is now shown to participate in skeletal muscle development. PMID- 12902329 TI - In vitro protein-DNA interactions at the human lamin B2 replication origin. AB - The complexity of mammalian origins of DNA replication has prevented, so far, the in vitro studies of the modalities of initiator protein binding and origin selection. We approached this problem by utilizing the human lamin B2 origin, wherein the precise start sites of replication initiation have been identified and known to be bound in vivo by the origin recognition complex (ORC). In order to analyze the in vitro interactions occurring at this origin, we have compared the DNA binding requirements and patterns of the human recombinant Orc4 with those of preparations of HeLa nuclear proteins containing the ORC complex. Here we show that both HsOrc4 alone and HeLa nuclear proteins recognize multiple sites within a 241-bp DNA sequence encompassing the lamin B2 origin. The DNA binding activity of HeLa cells requires the presence of ORC and can be reproduced in the absence of all the other proteins known to be recruited to origins by ORC. Both HsOrc4 alone and HeLa nuclear proteins exhibit cooperative and ATP-independent binding. This binding covers nucleotides 3853-3953 and then spreads outward. Because this region contains the start sites of DNA synthesis as well as the area protected in vivo and preserves protein binding capacity in vitro after removal of a fraction of the protected region, we suggest that it could contain the primary binding site. Thus the in vitro approach points to the sequence requirements for ORC binding as a key element for origin recognition. PMID- 12902330 TI - Helix 8 of the leukotriene B4 receptor is required for the conformational change to the low affinity state after G-protein activation. AB - Recent studies have revealed that G-protein-coupled receptors contain a putative cytoplasmic helical domain, helix 8. Leukotriene B4 (LTB4) receptor 1 derivatives with truncated or mutated helix 8 showed much higher LTB4 binding than wild-type (WT) receptors. Similar to the WT receptor, LTB4 promoted guanosine 5'-3-O (thio)triphosphate (GTPgammaS) binding in these mutants. Unlike the WT receptor, however, the addition of GTPgammaS did not inhibit LTB4 binding to the mutant receptors. Scatchard analyses revealed that mutants maintained high affinity for LTB4, even in the presence of excess GTPgammaS. Consistently, mutant receptors showed a more prolonged Ca2+ mobilization and cellular metabolic activation than the WT receptor. From mutational studies and three-dimensional modeling based on the structure of bovine rhodopsin, we conclude that the helix 8 of LTB4 receptor 1 plays an important role in the conformational change of the receptor to the low affinity state after G-protein activation, possibly by sensing the status of coupling Galpha subunits as GTP-bound. PMID- 12902331 TI - Complete reversal of coenzyme specificity by concerted mutation of three consecutive residues in alcohol dehydrogenase. AB - Gastric tissues from amphibian Rana perezi express the only vertebrate alcohol dehydrogenase (ADH8) that is specific for NADP(H) instead of NAD(H). In the crystallographic ADH8-NADP+ complex, a binding pocket for the extra phosphate group of coenzyme is formed by ADH8-specific residues Gly223-Thr224-His225, and the highly conserved Leu200 and Lys228. To investigate the minimal structural determinants for coenzyme specificity, several ADH8 mutants involving residues 223 to 225 were engineered and kinetically characterized. Computer-assisted modeling of the docked coenzymes was also performed with the mutant enzymes and compared with the wild-type crystallographic binary complex. The G223D mutant, having a negative charge in the phosphate-binding site, still preferred NADP(H) over NAD(H), as did the T224I and H225N mutants. Catalytic efficiency with NADP(H) dropped dramatically in the double mutants, G223D/T224I and T224I/H225N, and in the triple mutant, G223D/T224I/H225N (kcat/KmNADPH = 760 mm-1 min-1), as compared with the wild-type enzyme (kcat/KmNADPH = 133330 mm-1 min-1). This was associated with a lower binding affinity for NADP+ and a change in the rate limiting step. Conversely, in the triple mutant, catalytic efficiency with NAD(H) increased, reaching values (kcat/KmNADH = 155000 mm-1 min-1) similar to those of the wild-type enzyme with NADP(H). The complete reversal of ADH8 coenzyme specificity was therefore attained by the substitution of only three consecutive residues in the phosphate-binding site, an unprecedented achievement within the ADH family. PMID- 12902332 TI - Roles for inhibitory interactions in the use of the -10 promoter element by sigma 70 holoenzyme. AB - A panel of seven -10 region DNA mutants was tested for holoenzyme binding against a panel of 13 region 2 mutants of sigma 70. No patterns were noticed that would indicate unique interactions between individual amino acids and individual -10 region bases. Instead, certain amino acid substitutions led to increased holoenzyme binding to DNA, implying that the wild type interactions are associated with an inhibitory component. These inhibitory interactions were stronger on DNA containing non-consensus sequences, like those of typical promoters. In addition, the DNA segment downstream from the -10 element was also inhibitory to binding when in duplex form but stimulated binding when in single strand form. Overall, the data suggest that -10 region duplex recognition and melting have a large component of overcoming unfavorable protein:DNA base interactions, particularly when the bases are non-consensus, and that this contributes to setting physiologically appropriate variations in transcription rate. PMID- 12902333 TI - The expression of the thyroid-stimulating hormone (TSH) receptor and the cAMP dependent protein kinase RII beta regulatory subunit confers TSH-cAMP-dependent growth to mouse fibroblasts. AB - TSH activates its specific receptor in thyroid cells and induces cAMP, a robust stimulator of thyroid cell proliferation. Conversely, cAMP is a potent inhibitor of growth in mouse fibroblasts. To dissect the signals mediating cAMP-dependent growth, we have expressed in mouse fibroblasts the human thyrotropin receptor (TSHR) or a constitutively active mutant, under the control of the tetracyclin promoter. Both TSHR and cAMP levels were modulated by tetracyclin. In the presence of serum, activation of TSHR by TSH induced growth arrest. In the absence of serum, cells expressing TSHR stimulated with TSH, replicated their DNA, but underwent apoptosis. Co-expression of cAMP-dependent protein kinase (PKA) regulatory subunit type II (RIIbeta) inhibited apoptosis and stimulated the growth of cells only in the presence of TSH. Expression of RIIbeta-PKA, in the absence of TSHR, induced apoptosis, which was reversed by cAMP. Growth, stimulated by TSHR-RIIbeta-PKA in mouse fibroblasts, was also dependent on Rap1 activity, indicating cAMP-dependent growth in thyroid cells. As for the molecular mechanism underlying these effects, we found that in normal fibroblasts, TSH induced AKT and ERK1/2 only in cells expressing TSHR and RII. Similarly, activation of TSHR increased cAMP levels greatly, but was unable to stimulate CREB phosphorylation and transcription of cAMP-induced genes in the absence of RII. These data provide a simple explanation for the anti-proliferative and proliferative effects of cAMP in different cell types and indicate that RII-PKAII complements TSHR action by stably propagating robust cAMP signals in cell compartments. PMID- 12902334 TI - Evidence that intracellular beta1-2 mannan is a virulence factor in Leishmania parasites. AB - The protozoan parasite Leishmania mexicana proliferates within macrophage phagolysosomes in the mammalian host. In this study we provide evidence that a novel class of intracellular beta1-2 mannan oligosaccharides is important for parasite survival in host macrophages. Mannan (degree of polymerization 4-40) is expressed at low levels in non-pathogenic promastigote stages but constitutes 80 and 90% of the cellular carbohydrate in the two developmental stages that infect macrophages, non-dividing promastigotes, and lesion-derived amastigotes, respectively. Mannan is catabolized when parasites are starved of glucose, suggesting a reserve function, and developmental stages having low mannan levels or L. mexicana GDPMP mutants lacking all mannose molecules are highly sensitive to glucose starvation. Environmental stresses, such as mild heat shock or the heat shock protein-90 inhibitor, geldanamycin, that trigger the differentiation of promastigotes to amastigotes, result in a 10-25-fold increase in mannan levels. Developmental stages with low mannan levels or L. mexicana mutants lacking mannan do not survive heat shock and are unable to differentiate to amastigotes or infect macrophages in vitro. In contrast, a L. mexicana mutant deficient only in components of the mannose-rich surface glycocalyx differentiates normally and infects macrophages in vitro. Collectively, these data provide strong evidence that mannan accumulation is important for parasite differentiation and survival in macrophages. PMID- 12902335 TI - A specific role of the yeast mitochondrial carriers MRS3/4p in mitochondrial iron acquisition under iron-limiting conditions. AB - The yeast genes MRS3 and MRS4 encode two members of the mitochondrial carrier family with high sequence similarity. To elucidate their function we utilized genome-wide expression profiling and found that both deletion and overexpression of MRS3/4 lead to up-regulation of several genes of the "iron regulon." We therefore analyzed the two major iron-utilizing processes, heme formation and Fe/S protein biosynthesis in vivo, in organello (intact mitochondria), and in vitro (mitochondrial extracts). Radiolabeling of yeast cells with 55Fe revealed a clear correlation between MRS3/4 expression levels and the efficiency of these biosynthetic reactions indicating a role of the carriers in utilization and/or transport of iron in vivo. Similar effects on both heme formation and Fe/S protein biosynthesis were seen in organello using mitochondria isolated from cells grown under iron-limiting conditions. The correlation between MRS3/4 expression levels and the efficiency of the two iron-utilizing processes was lost upon detergent lysis of mitochondria. As no significant changes in the mitochondrial membrane potential were observed upon overexpression or deletion of MRS3/4, our results suggest that Mrs3/4p carriers are directly involved in mitochondrial iron uptake. Mrs3/4p function in mitochondrial iron transport becomes evident under iron-limiting conditions only, indicating that the two carriers do not represent the sole system for mitochondrial iron acquisition. PMID- 12902336 TI - Enzymatic E-colicins bind to their target receptor BtuB by presentation of a small binding epitope on a coiled-coil scaffold. AB - Toxins and viruses often initiate their attacks by binding to specific proteins on the surfaces of target cells. Bacterial toxins (e.g. bacteriocins) and viruses (bacteriophages) targeting Gram-negative bacteria typically bind to outer membrane proteins. Bacterial E-colicins target Escherichia coli by binding to the outer membrane cobalamin transporter BtuB. Colicins are tripartite molecules possessing receptor-binding, translocation, and toxin domains connected by long coiled-coil alpha-helices. Surprisingly, the crystal structure of colicin E3 does not possess a recognizable globular fold in its receptor-binding domain. We hypothesized that the binding epitope of enzymatic E-colicins is a short loop connecting the two alpha-helices that comprise the coiled-coil region and that this flanking coiled-coil region serves to present the loop in a binding-capable conformation. To test this hypothesis, we designed and synthesized a 34-residue peptide (E-peptide-1) corresponding to residues Ala366-Arg399 of the helix-loop helix region of colicin E3. Cysteines placed near the ends of the peptide (I372C and A393C) enabled crosslinking for reduction of conformational entropy and formation of a peptide structure that would present the loop epitope. A fluorescent analog was also made for characterization of binding by measurement of fluorescence polarization. Our analysis shows the following. (i). E-peptide-1 is predominantly random coil in aqueous solution, but disulfide bond formation increases its alpha-helical content in both aqueous buffer and solvents that promote helix formation. (ii). Fluorescein-labeled E-peptide-1 binds to purified BtuB in a calcium-dependent manner with a Kd of 43.6 +/- 4.9 nm or 2370 +/- 670 nm in the presence or absence of calcium, respectively. (iii). In the presence of calcium, cyanocobalamin (CN-Cbl) displaces E-peptide-1 with a nanomolar inhibition constant (Ki = 78.9 +/- 5.6 nm). We conclude that the BtuB binding sites for cobalamins and enzymatic E-colicins are overlapping but inequivalent and that the distal loop and (possibly) the short alpha-helical flanking regions are sufficient for high affinity binding. PMID- 12902337 TI - KCl cotransport is an important modulator of human cervical cancer growth and invasion. AB - Cervical cancer is a major world health problem for women, but the pathophysiology of this disease has received scant attention. Here we show that the growth and invasion of cervical cancer cells are strongly linked the expression and activity of the KCl cotransporter (KCC), an important regulator of the ionic and cellular osmotic homeostasis. Functional assays of KCl cotransport activation by osmotic swelling, staurosporine, and N-ethylmaleimide indicate that removal of the N-terminal 117 amino acids from KCC1 produces a dominant-negative loss-of-function phenotype for KCl cotransport in human cervical cancer cells. The capability for regulatory volume decrease is much attenuated in the loss-of function KCC mutant cervical cancer cells. The loss-of-function KCC mutant cervical cancer cells exhibit inhibited cell growth accompanied by decreased activity of the cell cycle gene products retinoblastoma and cdc2 kinase. Reduced cellular invasiveness is in parallel by reduced expression of alpha v beta 3 and alpha 6 beta 4 integrins, accompanied by decreased activity of matrix metalloproteinase 2 and 9. Inhibition of tumor growth in SCID mice confirms the crucial role of KCC in promoting cervical cancer growth and invasion. Thus, blockade of KCl cotransport may be a useful therapeutic adjunctive strategy to retard or prevent cervical cancer invasion. PMID- 12902338 TI - Identification of glucocorticoid receptor domains involved in transrepression of transforming growth factor-beta action. AB - The transforming growth factor-beta (TGF-beta) and glucocorticoid signaling pathways interact both positively and negatively in regulating a variety of physiological and pathologic processes. We previously reported that liganded glucocorticoid receptor (GR) repressed TGF-beta induction of human plasminogen activator inhibitor-1 gene transcription by directly targeting the transcriptional activation function of Smad3. To identify the domain(s) in the glucocorticoid receptor involved in this repression, we have examined the ability of various GR truncation, deletion, and substitution mutants to repress TGF-beta transactivation in Hep3B human hepatoma cells that lack functional endogenous GR. Partial deletions in the ligand-binding domain (LBD), including the tau2 and tauc regions, greatly reduced or eliminated GR repression, whereas deletion of the N terminal AF1 (tau1) domain and substitution mutations in the DNA-binding domain had little or no effect. Liganded androgen receptor repressed TGF-beta transactivation, whereas mineralocorticoid receptor did not, and studies with rat GR-mineralocorticoid receptor chimeras confirmed that the GR C-terminal domains were required for repression. RU486, a strong antagonist of transactivation by GR, partially reversed repression by wild type GR. Co-immunoprecipitation experiments in Hep3B cells indicated that physical interaction between GR and Smad3 is necessary but not sufficient for repression. Physical interaction required activation of Smad3 by TGF-beta but not dexamethasone binding to GR. Glutathione S-transferase pull-down assays demonstrated that several regions of the LBD could mediate GR-Smad3 physical interaction. We conclude that the LBD of GR, but not the DNA-binding domain or the N-terminal activation domain, is required for GR-mediated transrepression of TGF-beta transactivation. PMID- 12902339 TI - Kinetic and spectroscopic characterization of a hydroperoxy compound in the reaction of native myoglobin with hydrogen peroxide. AB - The reaction of metmyoglobin with H2O2 was investigated in a pH range between 8.5 and 6.0 with the aid of stopped flow-rapid scan and rapid freezing-EPR techniques. Singular value decomposition analyses of the stopped flow data at pH 8.5 revealed that a spectral species previously unknown accumulated during the reaction and exhibited a Soret absorption maximum at >/=423 nm. In the EPR experiments, the new species exhibited a set of g values at 2.32, 2.19, and 1.94, indicating that the species was assignable to a ferric hydroperoxy (Fe(III)[O-O H]-) compound. In contrast, the hydroperoxy compound scarcely accumulated in the reaction at pH 6.0, and the dominant intermediate species accumulated was compound I, which was derived from the oxygen-oxygen bond cleavage of the hydroperoxy compound. The accumulated amount of the hydroperoxy compound relative to compound I showed a pH dependence with an apparent pKa (pKaapp) from 6.95 to 7.27 depending on the metmyoglobins examined. This variation in pKaapp paralleled that in pKa of the acid-alkaline transition (pKaAB) of metmyoglobins, suggesting that the accumulation of hydroperoxy compound is controlled by the distal histidine. We propose that the H2O2 activation by metmyoglobin is promoted at the acidic condition due to the imidazolium form of the distal histidine, and we further propose that the controlled protonation state of the distal histidine is important for the facile O-O bond cleavage in heme peroxidases. PMID- 12902340 TI - Fusion of lamellar body with plasma membrane is driven by the dual action of annexin II tetramer and arachidonic acid. AB - Annexin II has been implicated in membrane fusion during the exocytosis of lamellar bodies from alveolar epithelial type II cells. Most previous studies were based on the fusion assays by using model membranes. In the present study, we investigated annexin II-mediated membrane fusion by using isolated lamellar bodies and plasma membrane as determined by the relief of octadecyl rhodamine B (R18) self-quenching. Immunodepletion of annexin II from type II cell cytosol reduced its fusion activity. Purified annexin II tetramer (AIIt) induced the fusion of lamellar bodies with the plasma membrane in a dose-dependent manner. This fusion is Ca2+-dependent and is highly specific to AIIt because other annexins (I and II monomer, III, IV, V, and VI) were unable to induce the fusion. Modification of the different functional residues of AIIt by N-ethylmaleimide, nitric oxide, or peroxynitrite abolished AIIt-mediated fusion. Arachidonic acid enhanced AIIt-mediated fusion and reduced its Ca2+ requirement to an intracellularly achievable level. This effect is due to membrane-bound arachidonic acid, not free arachidonic acid. Other fatty acids including linolenic acid, palmitoleic acid, myristoleic acid, stearic acid, palmitic acid, and myristic acid had little effect. AIIt-mediated fusion was suppressed by the removal of arachidonic acid from lamellar body and plasma membrane using bovine serum albumin. The addition of arachidonic acid back to the arachidonic acid depleted membranes restored its fusion activity. Our results suggest that the fusion between lamellar bodies with the plasma membrane is driven by the synergistic action of AIIt and arachidonic acid. PMID- 12902342 TI - Capacitative calcium entry induces hippocampal long term potentiation in the absence of presenilin-1. AB - Presenilins, whose mutant forms are the most common cause of early onset familial Alzheimer's disease, are involved in two very distinct processes: (i) proteolytic activity as gamma-secretase acting on amyloid precursor protein to produce amyloid peptides and (ii) storage of Ca2+ in the endoplasmic reticulum (ER). In particular, absence of presenilin-1 (PS1) was claimed to potentiate capacitative calcium entry (CCE), i.e. the mechanism of replenishment of ER Ca2+ stores. However, until now, evidence in favor of the latter role has been obtained only in isolated or cultured cells and not on neurons in situ. Here, we studied the strength of the synapses between Schaffer's collaterals and CA1 neurons in hippocampal slices when they were submitted first to Ca(2+)-free medium containing thapsigargin and subsequently to normal artificial cerebrospinal fluid, a procedure known to trigger CCE. We demonstrate that Ca2+ influx via the CCE mechanism is sufficient to trigger robust long term potentiation of the synapses in hippocampal slices from transgenic mice with a postnatal, neuron specific ablation of PS1, but remarkably not from wild-type mice. Our data establish for the first time in neurons confined in normal neuronal networks that PS1 acts on the refilling mechanism of ER Ca2+ stores. PMID- 12902341 TI - Structure of the HERG K+ channel S5P extracellular linker: role of an amphipathic alpha-helix in C-type inactivation. AB - The HERG K+ channel has very unusual kinetic behavior that includes slow activation but rapid inactivation. These features are critical for normal cardiac repolarization as well as in preventing lethal ventricular arrhythmias. Mutagenesis studies have shown that the extracellular peptide linker joining the fifth transmembrane domain to the pore helix is critical for rapid inactivation of the HERG K+ channel. This peptide linker is also considerably longer in HERG K+ channels, 40 amino acids, than in most other voltage-gated K+ channels. In this study we show that a synthetic 42-residue peptide corresponding to this linker region of the HERG K+ channel does not have defined structural elements in aqueous solution; however, it displays two well defined helical regions when in the presence of SDS micelles. The helices correspond to Trp585-Ile593 and Gly604 Tyr611 of the channel. The Trp585-Ile593 helix has distinct hydrophilic and hydrophobic surfaces. The Gly604-Tyr611 helix corresponds to an N-terminal extension of the pore helix. Electrophysiological studies of HERG currents following application of exogenous S5P peptides show that the amphipathic helix in the S5P linker interacts with the pore region of the channel in a voltage dependent manner. PMID- 12902343 TI - Ras signaling and transcriptional synergy at a flexible Ets-1/Pit-1 composite DNA element is defined by the assembly of selective activation domains. AB - Pit-1 and Ets-1 binding to a composite element synergistically activates and targets Ras-mitogen-activated protein kinase signaling to the rat prolactin promoter. These transcriptional responses appear to depend on three molecular features: organization of the Ets-1/Pit-1 composite element, physical interaction of these two factors via the Pit-1 homeodomain (amino acids 199-291) and the Ets 1 regulatory III domain (amino acids 190-257), and assembly of their transcriptional activation domains (TADs). Here we show that the organization of the Ets-1/Pit-1 composite element tolerates significant flexibility with regard to Ras stimulation and synergy. Specifically, the putative monomeric Pit-1 binding site can be substituted with bona fide binding sites for either a Pit-1 monomer or dimer, and these sites tolerated a separation of 28 bp. Additionally, we show that the physical interaction of Ets-1 and Pit-1 is not required for Ras responsiveness or synergy because block mutations of the Pit-1 interaction surface in Ets-1, which reduced Ets-1/Pit-1 binding in vitro, did not significantly affect Ets-1 stimulation of Ras responsiveness or synergy. We also show differential use of distinct TAD subtypes and Pit-1 TAD subregions to mediate either synergy or Ras responsiveness. Specifically, TADs from Gal4, VP16, or Ets-2 regulatory III domain linked to Ets-1 DNA binding domain constructs restored synergy to these TAD/Ets-1 DNA binding domain fusions. Conversely, deletion of the defined Pit-1 TAD (amino acids 2-80) retained synergy, but not Ras responsiveness. Consequently, we further defined the Pit-1 amino-terminal TAD into region 1 (R1, amino acids 2-45) and region 2 (R2, amino acids 46-80). R1 appears to regulate basal and synergistic responses, whereas the Ras response was mapped to R2. In summary, Ras responsiveness and Pit-1/Ets-1 synergy are mediated through the assembly of distinct TADs at a flexible composite element, indicating that different mechanisms underlie these two transcriptional responses and that the Pit-1 R2 subregion represents a novel, tissue-specific Ras-responsive TAD. PMID- 12902344 TI - Regulation of the discs large tumor suppressor by a phosphorylation-dependent interaction with the beta-TrCP ubiquitin ligase receptor. AB - The discs large (hDlg) tumor suppressor is intimately involved in the control of cell contact, polarity, and proliferation by interacting with several components of the epithelial junctional complex and with the APC tumor suppressor protein. In epithelial cells, hDlg protein stability is regulated through the ubiquitin proteasome pathway: hDlg is actively degraded in isolated cells, whereas it accumulates upon cell-cell contact. During neoplastic transformation of epithelial cells, loss of the differentiated morphology and progression toward a metastatic phenotype correlate with down-regulation of hDlg levels and loss of contact-dependent stabilization. Here we show that upon hyperphosphorylation, hDlg interacts with the beta-TrCP ubiquitin ligase receptor through a DSGLPS motif within its Src homology 3 domain. As a consequence, overexpression of beta TrCP enhances ubiquitination of Dlg protein and decreases its stability, whereas a dominant negative beta-TrCP mutant inhibits this process. Furthermore, a mutant Dlg protein that is unable to bind beta-TrCP displays a higher protein stability and is insensitive to beta-TrCP. Using RNA interference, we also demonstrate that endogenous beta-TrCP regulates hDlg protein levels in epithelial cells. Finally, we show that beta-TrCP selectively induces the degradation of the membrane cytoplasmic pool, without affecting the nuclear pool of hDlg. PMID- 12902345 TI - The Y181C substitution in 3'-azido-3'-deoxythymidine-resistant human immunodeficiency virus, type 1, reverse transcriptase suppresses the ATP-mediated repair of the 3'-azido-3'-deoxythymidine 5'-monophosphate-terminated primer. AB - Resistance to zidovudine (3'-azido-3'-deoxythymidine, AZT) by the human immunodeficiency virus, type 1, requires multiple amino acid substitutions such as D67N/K70R/T215F/K219Q in the viral reverse transcriptase (RT). In this background of AZT resistance, additional "suppressive" substitutions such as Y181C restore sensitivity to AZT. In order to characterize the mechanism of this AZT resistance suppression, the Y181C substitution was introduced into both wild type and AZT-resistant reverse transcriptase. The introduction of the Y181C substitution suppresses the increased repair (or unblocking) of the AZTMP terminated primer provided by the AZT resistance substitutions in RT using either DNA or RNA templates, independently from the RT RNase H activity. Contrary to wild-type RT, the low level of unblocking activity is not due to inhibition by the next correct nucleotide binding to the RT/AZTMP-terminated primer complex. When Y181C is added to the AZT resistance substitutions, ATP binds with less affinity to the AZTMP-terminated primer-RT binary complex. These results provide an insight into one possible molecular mechanism of re-sensitization of AZT resistant viruses by suppressive substitutions. PMID- 12902346 TI - Transcriptional down-regulation of peroxisome numbers affects selective peroxisome degradation in Hansenula polymorpha. AB - We have isolated and characterized a novel transcription factor of Hansenula polymorpha that is involved in the regulation of peroxisomal protein levels. This protein, designated Mpp1p, belongs to the family of Zn(II)2Cys6 proteins. In cells deleted for the function of Mpp1p the levels of various proteins involved in peroxisome biogenesis (peroxins) and function (enzymes) are reduced compared with wild type or, in the case of the matrix protein dihydroxyacetone synthase, fully absent. Also, upon induction of mpp1 cells on methanol, the number of peroxisomes was strongly reduced relative to wild type cells and generally amounted to one organelle per cell. Remarkably, this single organelle was not susceptible to selective peroxisome degradation (pexophagy) and remained unaffected during exposure of methanol-induced cells to excess glucose conditions. We show that this mechanism is a general phenomenon in H. polymorpha in the case of cells that contain only a single peroxisome. PMID- 12902348 TI - Lipopolysaccharide stimulates mitochondrial biogenesis via activation of nuclear respiratory factor-1. AB - Exposure to bacterial lipopolysaccharide (LPS) in vivo damages mitochondrial DNA (mtDNA) and interferes with mitochondrial transcription and oxidative phosphorylation (OXPHOS). Because this damage accompanies oxidative stress and is reversible, we postulated that LPS stimulates mtDNA replication and mitochondrial biogenesis via expression of factors responsive to reactive oxygen species, i.e. nuclear respiratory factor-1 (NRF-1) and mitochondrial transcription factor-A. In testing this hypothesis in rat liver, we found that LPS induces NRF-1 protein expression and activity accompanied by mRNA expression for mitochondrial transcription factor-A, mtDNA polymerase gamma, NRF-2, and single-stranded DNA binding protein. These events restored the loss in mtDNA copy number and OXPHOS gene expression caused by LPS and increased hepatocyte mitotic index, nuclear cyclin D1 translocation, and phosphorylation of pro-survival kinase, Akt. Thus, NRF-1 was implicated in oxidant-mediated mitochondrial biogenesis to provide OXPHOS for proliferation. This implication was tested in novel mtDNA-deficient cells generated from rat hepatoma cells that overexpress NRF-1. Depletion of mtDNA (rhoo clones) diminished oxidant production and caused loss of NRF-1 expression and growth delay. NRF-1 expression and growth were restored by exogenous oxidant exposure indicating that oxidative stress stimulates biogenesis in part via NRF-1 activation and corresponding to recovery events after LPS induced liver damage. PMID- 12902347 TI - Nerve growth factor-induced glutamate release is via p75 receptor, ceramide, and Ca(2+) from ryanodine receptor in developing cerebellar neurons. AB - Very little is known about the contribution of a low affinity neurotrophin receptor, p75, to neurotransmitter release. Here we show that nerve growth factor (NGF) induced a rapid release of glutamate and an increase of Ca2+ in cerebellar neurons through a p75-dependent pathway. The NGF-induced release occurred even in the presence of the Trk inhibitor K252a. The release caused by NGF but not brain derived neurotrophic factor was enhanced in neurons overexpressing p75. Further, after transfection of p75-small interfering RNA, which down-regulated the endogenous p75 expression, the NGF-induced release was inhibited, suggesting that the NGF-induced glutamate release was through p75. We found that the NGF increased Ca2+ was derived from the ryanodine-sensitive Ca2+ receptor and that the NGF-increased Ca2+ was essential for the NGF-induced glutamate release. Furthermore, scyphostatin, a sphingomyelinase inhibitor, blocked the NGF dependent Ca2+ increase and glutamate release, suggesting that a ceramide produced by sphingomyelinase was required for the NGF-stimulated Ca2+ increase and glutamate release. This action of NGF only occurred in developing neurons whereas the brain-derived neurotrophic factor-mediated Ca2+ increase and glutamate release was observed at the mature neuronal stage. Thus, we demonstrate that NGF-mediated neurotransmitter release via the p75-dependent pathway has an important role in developing neurons. PMID- 12902349 TI - Resveratrol-induced apoptosis is associated with Fas redistribution in the rafts and the formation of a death-inducing signaling complex in colon cancer cells. AB - Resveratrol, a polyphenol found in grape skin and various other food products, may function as a cancer chemopreventive agent for colon and other malignant tumors and possesses a chemotherapeutic potential through its ability to trigger apoptosis in tumor cells. The present study analyses the molecular mechanisms of resveratrol-induced apoptosis in colon cancer cells, with special attention to the role of the death receptor Fas in this pathway. We show that, in the 10-100 microm range of concentrations, resveratrol activates various caspases and triggers apoptosis in SW480 human colon cancer cells. Caspase activation is associated with accumulation of the pro-apoptotic proteins Bax and Bak that undergo conformational changes and relocalization to the mitochondria. Resveratrol does not modulate the expression of Fas and Fas-ligand (FasL) at the surface of cancer cells, and inhibition of the Fas/FasL interaction does not influence the apoptotic response to the molecule. Resveratrol induces the clustering of Fas and its redistribution in cholesterol and sphingolipid-rich fractions of SW480 cells, together with FADD and procaspase-8. This redistribution is associated with the formation of a death-inducing signaling complex (DISC). Transient transfection of either a dominant-negative mutant of FADD, E8, or MC159 viral proteins that interfere with the DISC function, decreases the apoptotic response of SW480 cells to resveratrol and partially prevents resveratrol-induced Bax and Bak conformational changes. Altogether, these results indicate that the ability of resveratrol to induce the redistribution of Fas receptor in membrane rafts may contribute to the molecule's ability to trigger apoptosis in colon cancer cells. PMID- 12902350 TI - Chromogranin B-induced secretory granule biogenesis: comparison with the similar role of chromogranin A. AB - The two major proteins of secretory granules of secretory cells, chromogranins A (CGA) and B (CGB), have previously been proposed to play key roles in secretory granule biogenesis. Recently, CGA was reported to play an on/off switch role for secretory granule biogenesis. In the present study we found CGB being more effective than CGA in inducing secretory granule formation in non-neuroendocrine NIH3T3 and COS-7 cells. The mean number of dense core granules formed/cell of CGA transfected NIH3T3 cells was 2.51, whereas that of CGB-transfected cells was 4.02, indicating the formation of 60% more granules in the CGB-transfected cells. Similarly, there were 55% more dense core granules formed in the CGB-transfected COS-7 cells than in the CGA-transfected cells. Moreover, transfection of CGA- and CGB-short interfering RNA (siRNA) into neuroendocrine PC12 cells not only decreased the amount of CGA and CGB expressed but also reduced the number of secretory granules by 41 and 78%, respectively, further suggesting the importance of CGB expression in secretory granule formation. PMID- 12902351 TI - Prostate cancer cell-adipocyte interaction: leptin mediates androgen-independent prostate cancer cell proliferation through c-Jun NH2-terminal kinase. AB - Prostate cancer is one of the leading causes of death among men in the United States, and acquisition of hormone resistance (androgen independence) by cancer cells is a fatal event during the natural history of prostate cancer. Obesity is another serious health problem and has been shown to be associated with prostate cancer. However, little is known about the molecular basis of this association. Here we show that factor(s) secreted from adipocytes stimulate prostate cancer cell proliferation. Leptin is one of the major adipose cytokines, and it controls body weight homeostasis through food intake and energy expenditure. We identify leptin as a novel growth factor in androgen-independent prostate cancer cell growth. Strikingly, leptin stimulates cell proliferation specifically in androgen independent DU145 and PC-3 prostate cancer cells but not in androgen-dependent LNCaP-FGC cells, although both cell types express functional leptin receptor isoforms. c-Jun NH2-terminal kinase (JNK) has been shown recently to play a crucial role in obesity and insulin resistance. Intriguingly, leptin induces JNK activation in androgen-independent prostate cancer cells, and the pharmacological inhibition of JNK blocked the leptin stimulation of androgen-independent prostate cancer cell proliferation. This suggests that JNK activation is required for leptin-mediated, androgen-independent prostate cancer cell proliferation. Furthermore, other cytokines produced by adipocytes and critical for body weight homeostasis cooperate with leptin in androgen-independent prostate cancer cell proliferation: interleukin-6 and insulin-like growth factor I demonstrate additive and synergistic effects on the leptin stimulation of androgen independent prostate cancer cell proliferation, respectively. Therefore, adipose cytokines, as well as JNK, are key mediators between obesity and hormone resistant prostate cancer and could be therapeutic targets. PMID- 12902352 TI - Analysis of DNA replication intermediates suggests mechanisms of repeat sequence expansion. AB - We previously developed a system to investigate the mechanism of repeat sequence expansion during eukaryotic Okazaki fragment processing. Upstream and downstream primers were annealed to a complementary template to overlap across a CAG repeat region. Annealing by the competing primers lead to structural intermediates that ligated to expand the repeat segment. When an equal number of repeats overlapped on the upstream and downstream primers, a 2-fold expansion was expected, but no expansion occurred. We show here that such substrates do not expand irrespective of their repeat length. To reveal mechanism, we tested different hairpin loop intermediates expected to form and facilitate ligation. Substrates configured to form large loops in either the upstream or downstream primer alone allowed expansion. Large or small fixed position single loops allowed expansion when located at least six nucleotides up- or downstream of the nick. Fixed loops in both primers, simulating a double loop intermediate, allowed expansion as long as each loop was nine nucleotides from the nick. Thus, neither the double loop configuration required to form with equal length overlaps nor the large single loop configuration are fundamental structural impediments to expansion. We propose a model for the expansion mechanism based on the relative stabilities of single loop, double loop, hairpin, and flap intermediates that is consistent with the observed expansion efficiency of equal and unequal overlap substrates. The model suggests that the equilibrium concentration of double loop intermediates is so vanishingly small that they are not likely contributors to sequence expansion. PMID- 12902353 TI - Characterization of 2'-fluoro-RNA aptamers that bind preferentially to disease associated conformations of prion protein and inhibit conversion. AB - We have isolated artificial ligands or aptamers for infectious prions in order to investigate conformational aspects of prion pathogenesis. The aptamers are 2' fluoro-modified RNA produced by in vitro selection from a large, randomized library. One of these ligands (aptamer SAF-93) had more than 10-fold higher affinity for PrPSc than for recombinant PrPC and inhibited the accumulation of PrPres in near physiological cell-free conversion assay. To understand the molecular basis of these properties and to distinguish specific from non-specific aptamer-PrP interactions, we studied deletion mutants of bovine PrP in denatured, alpha-helix-rich and beta-sheet-rich forms. We provide evidence that, like scrapie-associated fibrils (SAF), the beta-oligomer of PrP bound to SAF-93 with at least 10-fold higher affinity than did the alpha-form. This differential affinity could be explained by the existence of two binding sites within the PrP molecule. Site 1 lies within residues 23-110 in the unstructured N terminus and is a nonspecific RNA binding site found in all forms of PrP. The region between residue 90 and 110 forms a hinge region that is occluded in the alpha-rich form of PrP but becomes exposed in the denatured form of PrP. Site 2 lies in the region C-terminal of residue 110. This site is beta-sheet conformation-specific and is not recognized by control RNAs. Taken together, these data provide for the first time a specific ligand for a disease conformation-associated site in a region of PrP critical for conformational conversion. This aptamer could provide tools for the further analysis of the processes of PrP misfolding during prion disease and leads for the development of diagnostic and therapeutic approaches to TSEs. PMID- 12902354 TI - Taking aim at gun violence: tracing bullets and guns to sideline scofflaw dealers. PMID- 12902355 TI - Programs target youth violence prevention. PMID- 12902356 TI - In the wake of tragedy: studies track psychological response to mass violence. PMID- 12902359 TI - Premature termination of clinical trials. PMID- 12902358 TI - The World Trade Center disaster and intrauterine growth restriction. PMID- 12902360 TI - Accuracy of a local surveillance system for early detection of emerging infectious disease. PMID- 12902361 TI - Bronchiolitis obliterans in a survivor of a chemical weapons attack. PMID- 12902362 TI - Sudden death due to neck blows among amateur hockey players. PMID- 12902363 TI - A mental health intervention for schoolchildren exposed to violence: a randomized controlled trial. AB - CONTEXT: No randomized controlled studies have been conducted to date on the effectiveness of psychological interventions for children with symptoms of posttraumatic stress disorder (PTSD) that has resulted from personally witnessing or being personally exposed to violence. OBJECTIVE: To evaluate the effectiveness of a collaboratively designed school-based intervention for reducing children's symptoms of PTSD and depression that has resulted from exposure to violence. DESIGN: A randomized controlled trial conducted during the 2001-2002 academic year. SETTING AND PARTICIPANTS: Sixth-grade students at 2 large middle schools in Los Angeles who reported exposure to violence and had clinical levels of symptoms of PTSD. INTERVENTION: Students were randomly assigned to a 10-session standardized cognitive-behavioral therapy (the Cognitive-Behavioral Intervention for Trauma in Schools) early intervention group (n = 61) or to a wait-list delayed intervention comparison group (n = 65) conducted by trained school mental health clinicians. MAIN OUTCOME MEASURES: Students were assessed before the intervention and 3 months after the intervention on measures assessing child reported symptoms of PTSD (Child PTSD Symptom Scale; range, 0-51 points) and depression (Child Depression Inventory; range, 0-52 points), parent-reported psychosocial dysfunction (Pediatric Symptom Checklist; range, 0-70 points), and teacher-reported classroom problems using the Teacher-Child Rating Scale (acting out, shyness/anxiousness, and learning problems; range of subscales, 6-30 points). RESULTS: Compared with the wait-list delayed intervention group (no intervention), after 3 months of intervention students who were randomly assigned to the early intervention group had significantly lower scores on symptoms of PTSD (8.9 vs 15.5, adjusted mean difference, - 7.0; 95% confidence interval [CI], - 10.8 to - 3.2), depression (9.4 vs 12.7, adjusted mean difference, - 3.4; 95% CI, - 6.5 to - 0.4), and psychosocial dysfunction (12.5 vs 16.5, adjusted mean difference, - 6.4; 95% CI, -10.4 to -2.3). Adjusted mean differences between the 2 groups at 3 months did not show significant differences for teacher-reported classroom problems in acting out (-1.0; 95% CI, -2.5 to 0.5), shyness/anxiousness (0.1; 95% CI, -1.5 to 1.7), and learning (-1.1, 95% CI, -2.9 to 0.8). At 6 months, after both groups had received the intervention, the differences between the 2 groups were not significantly different for symptoms of PTSD and depression; showed similar ratings for psychosocial function; and teachers did not report significant differences in classroom behaviors. CONCLUSION: A standardized 10-session cognitive-behavioral group intervention can significantly decrease symptoms of PTSD and depression in students who are exposed to violence and can be effectively delivered on school campuses by trained school-based mental health clinicians. PMID- 12902364 TI - Exposure to terrorism, stress-related mental health symptoms, and coping behaviors among a nationally representative sample in Israel. AB - CONTEXT: The terrorist attacks on Israeli society have been ongoing since September 2000. However, few studies have examined the impact of terrorism on nationally representative population samples, and no study has examined the psychological impact of ongoing terrorism in Israel. OBJECTIVES: To determine the level of exposure to terrorist attacks and the prevalence of traumatic stress related (TSR) symptoms, symptoms of posttraumatic stress disorder (PTSD), and sense of safety after 19 months of terrorism in Israel, and to identify correlates of the psychological sequelae and the modes of coping with the terrorism. DESIGN, SETTING, AND PARTICIPANTS: Telephone survey conducted April May 2002, using a strata sampling method, of 902 eligible households and a representative sample of 742 Israeli residents older than 18 years (82% contact rate) and a final participation of 512 (57%). MAIN OUTCOME MEASURES: Number of TSR symptoms, rates of those with symptom criteria for PTSD and acute stress disorder assessed by the Stanford Acute Stress Reaction Questionnaire, self reported feelings of depression, optimism, sense of safety, help-seeking, and modes of coping. RESULTS: Of 512 survey participants, 84 (16.4%) had been directly exposed to a terrorist attack and 191 (37.3%) had a family member or friend who had been exposed. Of 510 participants who responded to questions about TSR symptoms, 391 (76.7%) had at least 1 TSR symptom (mean, 4.0 [SD, 4.5]; range, 0-23; mean intensity, 0.8; range, 0-4). Symptom criteria for PTSD were met by 48 participants (9.4%) and criteria for acute stress disorder, by 1 participant; 299 (58.6%) reported feeling depressed. The majority of respondents expressed optimism about their personal future (421/512 [82.2%]) and the future of Israel (307/509 [66.8%]), and expressed self-efficacy with regard to their ability to function in a terrorist attack (322/431 [74.6%]). Most expressed a low sense of safety with respect to themselves (307/509 [60.4%]) and their relatives (345/507 [67.9%]). Few reported a need for professional help (27/506 [5.3%]). Female sex, sense of safety, and use of tranquilizers, alcohol, and cigarettes to cope were associated with TSR symptoms and symptom criteria for PTSD; level of exposure and objective risk were not. The most prevalent coping mechanisms were active information search about loved ones and social support. CONCLUSIONS: Considering the nature and length of the Israeli traumatic experience, the psychological impact may be considered moderate. Although the survey participants showed distress and lowered sense of safety, they did not develop high levels of psychiatric distress, which may be related to a habituation process and to coping mechanisms. PMID- 12902365 TI - A population-based study of inflicted traumatic brain injury in young children. AB - CONTEXT: Physical abuse is a leading cause of serious head injury and death in children aged 2 years or younger. The incidence of inflicted traumatic brain injury (TBI) in US children is unknown. OBJECTIVE: To determine the incidence of serious or fatal inflicted TBI in a defined US population of approximately 230 000 children aged 2 years or younger. DESIGN, SETTING, AND SUBJECTS: All North Carolina children aged 2 years or younger who were admitted to a pediatric intensive care unit or who died with a TBI in 2000 and 2001 were identified prospectively. Injuries were considered inflicted if accompanied by a confession or a medical and social service agency determination of abuse. MAIN OUTCOME MEASURE: Incidence of inflicted TBI. Multivariate logistic regression models were used to compare children with inflicted injuries with those with noninflicted injuries and with the general state population aged 2 years or younger. RESULTS: A total of 152 cases of serious or fatal TBI were identified, with 80 (53%) incurring inflicted TBI. The incidence of inflicted traumatic brain injury in the first 2 years of life was 17.0 (95% confidence interval [CI], 13.3-20.7) per 100 000 person-years. Infants had a higher incidence than children in the second year of life (29.7 [95% CI, 22.9-36.7] vs 3.8 [95% CI, 1.3-6.4] per 100 000 person years). Boys had a higher incidence than girls (21.0 [95% CI, 15.1-26.6] vs 13.0 [95% CI, 8.4-17.7] per 100 000 person-years). Relative to the general population, children who incurred an increased risk of inflicted injury were born to young mothers (< or =21 years), non-European American, or products of multiple births. CONCLUSIONS: In this population of North Carolina children, the incidence of inflicted TBI varied by characteristics of the injured children and their mothers. These data may be helpful for informing preventive interventions. PMID- 12902366 TI - Mental health and health-related quality of life among adult Latino primary care patients living in the United States with previous exposure to political violence. AB - CONTEXT: Although political violence continues in parts of Central America, South America, and Mexico, little is known about its relationship to the health of Latino immigrants living in the United States. OBJECTIVE: To determine (1) rates of exposure to political violence among Latino adult primary care patients who have immigrated to the United States from Central America, South America, and Mexico and its impact on mental health and health-related quality of life and (2) frequency of disclosure of political violence to primary care clinicians. DESIGN, SETTING, AND PARTICIPANTS: Two-stage cluster design survey of a systematic sample of Latino immigrant adults in 3 community-based primary care clinics in Los Angeles, conducted from July 2001 to February 2002. MAIN OUTCOME MEASURES: Reports of exposure to political violence in home country before immigrating to the United States and communication with clinicians about political violence; self-reported measures of health-related quality of life using the Medical Outcomes Study Short Form 36 (MOS SF-36); symptoms of depression, anxiety, and alcohol disorders using the Primary Care Evaluation of Mental Disorders (PRIME MD); and symptoms of posttraumatic stress disorder (PTSD) using the PTSD Checklist-Civilian Version (PCL-C). RESULTS: A total of 638 (69%) of 919 eligible patients participated. The nonresponse rates did not differ by age, sex, recruitment sites, or clinic sessions. In weighted analyses, 54% of participants reported political violence experiences in their home countries, including 8% who reported torture. Of those exposed to political violence, 36% had symptoms of depression and 18% had symptoms of PTSD vs 20% and 8%, respectively, among those not exposed to political violence. Controlling for age, sex, country, years lived in the United States, acculturation, income, health insurance status, and recruitment site in a subsample of 512 participants (56%), those who reported political violence exposure were more likely to meet symptom criteria for PTSD (adjusted odds ratio [AOR], 3.4; 95% confidence interval [CI], 1.4-8.4) and to have symptoms of depression (AOR, 2.8; 95% CI, 1.4-5.4) and symptoms of panic disorder (AOR, 4.8; 95% CI, 1.6-14.4) than participants not reporting political violence. Those exposed to political violence reported more chronic pain and role limitations due to physical problems, as well as worse physical functioning and lower perceptions of general health than those who were not exposed to political violence. Only 3% of the 267 patients who had experienced political violence reported ever telling a clinician about it after immigrating; none reported their current physician asking about political violence. CONCLUSION: Latino immigrants in primary care in Los Angeles have a high prevalence of exposure to political violence before immigrating to the United States and associated impairments in mental health and health-related quality of life. PMID- 12902367 TI - Factors associated with poor mental health among Guatemalan refugees living in Mexico 20 years after civil conflict. AB - CONTEXT: From 1981 to 2001, 46 000 refugees who fled the 36-year civil conflict in Guatemala for Chiapas, Mexico were under the protection of the United Nations High Commissioner for Refugees. OBJECTIVES: To estimate the prevalence of mental illness and factors associated with poor mental health of underserved Guatemalan refugee communities located in Chiapas, Mexico, since 1981 and to assess need for mental health services. DESIGN, SETTING, AND PARTICIPANTS: Cross-sectional survey of 183 households in 5 Mayan refugee camps in Chiapas representing an estimated 1546 residents (adults and children) conducted November-December 2000. MAIN OUTCOME MEASURES: Symptom criteria of Posttraumatic Stress Disorder (PTSD), anxiety, and depression as measured by the Harvard Trauma Questionnaire and Hopkins Symptom Checklist-25 (Hopkins-25). RESULTS: One adult (aged > or =16 years) per household (n = 170 respondents) who agreed to participate was included in the analysis, representing an estimated 93% of households. All respondents reported experiencing at least 1 traumatic event with a mean of 8.3 traumatic events per individual. Of the respondents, 20 (11.8%) had all symptom criteria for PTSD. Of the 160 who completed the Hopkins Symptom Checklist-25, 87 (54.4%) had anxiety symptoms and 62 (38.8%) had symptoms of depression. Witnessing the disappearance of family members (adjusted odds ratio [AOR], 4.58; 95% confidence interval [CI], 1.35-15.50), being close to death (AOR, 4.19, 95% CI, 1.03-17.00), or living with 9 to 15 persons in the same home (AOR, 3.69; 95% CI, 1.19-11.39) were associated with symptoms of PTSD. There was a protective factor found for lacking sufficient food (AOR, 0.08; 95% CI, 0.01-0.59). Elevated anxiety symptoms were associated with witnessing a massacre (AOR, 10.63; 95% CI, 4.31-26.22), being wounded (AOR, 3.22; 95% CI, 0.95-10.89), and experiencing 7 to 12 traumatic events (AOR, 2.67; 95% CI, 1.14-6.27) and 13 to 19 traumatic events (AOR, 2.26; 95% CI, 0.65-7.89). Elevated symptoms of depression were associated with being a woman (AOR, 3.64; 95% CI, 1.47-9.04), being widowed (AOR, 27.55; 95% CI, 2.54 299.27), being married (AOR, 1.93; 95% CI, 0.59-6.33), witnessing disappearances (AOR, 2.68; 95% CI, 1.16-6.19), experiencing 7 to 12 traumatic events (AOR, 1.57; 95% CI, 0.64-3.88), or experiencing 13 to 19 traumatic events (AOR, 7.44; 95% CI, 2.18-25.37). CONCLUSION: Psychiatric morbidity related to human rights violations, traumatic events, and refugee status was common among Guatemalan refugees surveyed 20 years after the Guatemalan civil conflict. PMID- 12902368 TI - Screening for risk of persistent posttraumatic stress in injured children and their parents. AB - CONTEXT: Injury, a leading health threat to children, is also a common cause of posttraumatic stress disorder (PTSD) in childhood. Most injured children with PTSD are not diagnosed or treated. OBJECTIVE: To develop a stand-alone screening tool for use by clinicians during acute trauma care to identify injured children and their parents who are at risk of significant, persistent posttraumatic stress symptoms. DESIGN: The Screening Tool for Early Predictors of PTSD (STEPP) was derived from a 50-item risk factor survey administered within 1 month of injury as part of a prospective cohort study of posttraumatic stress in injured children and their parents. Symptoms of PTSD were assessed at least 3 months after injury. SETTING: Urban, pediatric level I trauma center. PARTICIPANTS: A sample of 269 children aged 8 to 17 years admitted for treatment of traffic-related injuries between July 1999 and October 2001, and one parent per child, completed a risk factor survey assessing potential predictors of PTSD outcome. One hundred seventy one families (63%) completed a follow-up assessment. MAIN OUTCOME MEASURES: The Clinician-Administered PTSD Scale for Children and Adolescents and the PTSD Checklist served as criterion standards for child and parent outcomes, respectively. Positive cases were defined as those meeting criteria for at least subsyndromal PTSD with continuing impairment ("persistent traumatic stress"). RESULTS: The STEPP contains 4 dichotomous questions asked of the child, 4 asked of one parent, and 4 items obtained easily from the emergency medical record. STEPP sensitivity in predicting posttraumatic stress was 0.88 for children and 0.96 for parents, with negative predictive values of 0.95 for children and 0.99 for parents. The odds ratio for prediction of persistent traumatic stress was 6.5 (95% confidence interval [CI], 1.8-22.8) in children and 26.6 (95% CI, 3.5-202.1) in parents. CONCLUSIONS: The STEPP represents a new method to guide clinicians in making evidence-based decisions for the allocation of scarce mental health resources for traumatic stress. Its brevity and simple scoring rule suggest that it can be easily administered in the acute care setting. PMID- 12902369 TI - Death and injury from landmines and unexploded ordnance in Afghanistan. AB - CONTEXT: Afghanistan is one of the countries most affected by injuries due to landmines and unexploded ordnance. OBJECTIVE: To understand the epidemiological patterns and risk factors for injury due to landmines and unexploded ordnance. DESIGN AND SETTING: Analysis of surveillance data on landmine and unexploded ordnance injuries in Afghanistan collected by the International Committee of the Red Cross in 390 health facilities in Afghanistan. Surveillance data were used to describe injury trends, injury types, demographics, and risk behaviors of those injured and explosive types related to landmine and unexploded ordnance incidents. PARTICIPANTS: A total of 1636 individuals injured by landmines and unexploded ordnance, March 2001 through June 2002. RESULTS: Eighty-one percent of those injured were civilians, 91.6% were men and boys, and 45.9% were younger than 16 years. Children were more likely to be injured by unexploded ordnance (which includes grenades, bombs, mortar shells, and cluster munitions), whereas adults were injured mostly by landmines. The most common risk behaviors for children were playing and tending animals; for adults, these risk behaviors were military activity and activities of economic necessity (eg, farming, traveling). The case-fatality rate of 9.4% is probably underestimated because surveillance predominantly detects those who survive long enough to receive medical care. CONCLUSIONS: Landmine risk education should focus on hazards due to unexploded ordnance for children and on landmine hazards for adults and should address age specific risk behaviors. Expanding community-based and clinic-based reporting will improve the sensitivity and representativeness of surveillance. PMID- 12902370 TI - Challenges for communicable disease surveillance and control in southern Iraq, April-June 2003. AB - The recent war in Iraq presents significant challenges for the surveillance and control of communicable diseases. In early April 2003, the World Health Organization (WHO) sent a team of public health experts to Kuwait and a base was established in the southern Iraqi governorate of Basrah on May 3. We present the lessons learned from the communicable disease surveillance and control program implemented in the Basrah governorate in Iraq (population of 1.9 million) in April and May 2003, and we report communicable disease surveillance data through June 2003. Following the war, communicable disease control programs were disrupted, access to safe water was reduced, and public health facilities were looted. Rapid health assessments were carried out in health centers and hospitals to identify priorities for action. A Health Sector Coordination Group was organized with local and international health partners, and an early warning surveillance system for communicable disease was set up. In the first week of May 2003, physicians in hospitals in Basrah suspected cholera cases and WHO formed a cholera control committee. As of June 29, 2003, Iraqi hospital laboratories have confirmed 94 cases of cholera from 7 of the 8 districts of the Basrah governorate. To prevent the transmission of major communicable diseases, restoring basic public health and water/sanitation services is currently a top priority in Iraq. Lack of security continues to be a barrier for effective public health surveillance and response in Iraq. PMID- 12902371 TI - Clinical manifestations of sarin nerve gas exposure. PMID- 12902372 TI - Exhumation of mass graves in Iraq: considerations for forensic investigations, humanitarian needs, and the demands of justice. PMID- 12902373 TI - Posttraumatic symptoms and the complexity of responses to trauma. PMID- 12902374 TI - JAMA patient page. Inflicted brain injury in children. PMID- 12902375 TI - Tropheryma whipplei Twist: a human pathogenic Actinobacteria with a reduced genome. AB - The human pathogen Tropheryma whipplei is the only known reduced genome species (<1 Mb) within the Actinobacteria [high G+C Gram-positive bacteria]. We present the sequence of the 927303-bp circular genome of T. whipplei Twist strain, encoding 808 predicted protein-coding genes. Specific genome features include deficiencies in amino acid metabolisms, the lack of clear thioredoxin and thioredoxin reductase homologs, and a mutation in DNA gyrase predicting a resistance to quinolone antibiotics. Moreover, the alignment of the two available T. whipplei genome sequences (Twist vs. TW08/27) revealed a large chromosomal inversion the extremities of which are located within two paralogous genes. These genes belong to a large cell-surface protein family defined by the presence of a common repeat highly conserved at the nucleotide level. The repeats appear to trigger frequent genome rearrangements in T. whipplei, potentially resulting in the expression of different subsets of cell surface proteins. This might represent a new mechanism for evading host defenses. The T. whipplei genome sequence was also compared to other reduced bacterial genomes to examine the generality of previously detected features. The analysis of the genome sequence of this previously largely unknown human pathogen is now guiding the development of molecular diagnostic tools and more convenient culture conditions. PMID- 12902376 TI - Natural genomic design in Sinorhizobium meliloti: novel genomic architectures. AB - The complete nucleotide sequence of the genome of Sinorhizobium meliloti, the symbiont of alfalfa, was reported in 2001 by an international consortium of laboratories. The genome comprises a chromosome of 3.65 megabases (Mb) and two megaplasmids, pSymA and pSymB, of 1.35 Mb and 1.68 Mb, respectively. Based on the nucleotide sequence of the whole genome, we designed a pathway of consecutive rearrangements leading to novel genomic architectures. In a first step we obtained derivative strains containing two replicons; in a second step we obtained a strain containing the genetic information in one single replicon of 6.68 MB. From this last architecture we isolated revertants containing two replicons, and from these we could return to the original architecture showing the three replicons. We found that the relative frequency of excision of cointegrated replicons is higher at the site used for the cointegration than at other sites. This might conciliate two apparently opposed facts: the highly dynamic state of genomic architecture in S. meliloti and the common observation that different isolates and derived cellular clones of S. meliloti usually present the architecture of one chromosome and two distinct megaplasmids. Different aspects that must be considered to obtain full advantage of the strategy of natural genomic design are discussed. PMID- 12902377 TI - Comparative DNA sequence analysis of wheat and rice genomes. AB - The use of DNA sequence-based comparative genomics for evolutionary studies and for transferring information from model species to crop species has revolutionized molecular genetics and crop improvement strategies. This study compared 4485 expressed sequence tags (ESTs) that were physically mapped in wheat chromosome bins, to the public rice genome sequence data from 2251 ordered BAC/PAC clones using BLAST. A rice genome view of homologous wheat genome locations based on comparative sequence analysis revealed numerous chromosomal rearrangements that will significantly complicate the use of rice as a model for cross-species transfer of information in nonconserved regions. PMID- 12902378 TI - A gene recommender algorithm to identify coexpressed genes in C. elegans. AB - One of the most important uses of whole-genome expression data is for the discovery of new genes with similar function to a given list of genes (the query) already known to have closely related function. We have developed an algorithm, called the gene recommender, that ranks genes according to how strongly they correlate with a set of query genes in those experiments for which the query genes are most strongly coregulated. We used the gene recommender to find other genes coexpressed with several sets of query genes, including genes known to function in the retinoblastoma complex. Genetic experiments confirmed that one gene (JC8.6) identified by the gene recommender acts with lin-35 Rb to regulate vulval cell fates, and that another gene (wrm-1) acts antagonistically. We find that the gene recommender returns lists of genes with better precision, for fixed levels of recall, than lists generated using the C. elegans expression topomap. PMID- 12902379 TI - Allelic variation in gene expression is common in the human genome. AB - Variations in gene sequence and expression underlie much of human variability. Despite the known biological roles of differential allelic gene expression resulting from X-chromosome inactivation and genomic imprinting, a large-scale analysis of allelic gene expression in human is lacking. We examined allele specific gene expression of 1063 transcribed single-nucleotide polymorphisms (SNPs) by using Affymetrix HuSNP oligo arrays. Among the 602 genes that were heterozygous and expressed in kidney or liver tissues from seven individuals, 326 (54%) showed preferential expression of one allele in at least one individual, and 170 of those showed greater than fourfold difference between the two alleles. The allelic variation has been confirmed by real-time quantitative PCR experiments. Some of these 170 genes are known to be imprinted, such as SNRPN, IPW, HTR2A, and PEG3. Most of the differentially expressed genes are not in known imprinting domains but instead are distributed throughout the genome. Our studies demonstrate that variation of gene expression between alleles is common, and this variation may contribute to human variability. PMID- 12902380 TI - Decay rates of human mRNAs: correlation with functional characteristics and sequence attributes. AB - Although mRNA decay rates are a key determinant of the steady-state concentration for any given mRNA species, relatively little is known, on a population level, about what factors influence turnover rates and how these rates are integrated into cellular decisions. We decided to measure mRNA decay rates in two human cell lines with high-density oligonucleotide arrays that enable the measurement of decay rates simultaneously for thousands of mRNA species. Using existing annotation and the Gene Ontology hierarchy of biological processes, we assign mRNAs to functional classes at various levels of resolution and compare the decay rate statistics between these classes. The results show statistically significant organizational principles in the variation of decay rates among functional classes. In particular, transcription factor mRNAs have increased average decay rates compared with other transcripts and are enriched in "fast-decaying" mRNAs with half-lives <2 h. In contrast, we find that mRNAs for biosynthetic proteins have decreased average decay rates and are deficient in fast-decaying mRNAs. Our analysis of data from a previously published study of Saccharomyces cerevisiae mRNA decay shows the same functional organization of decay rates, implying that it is a general organizational scheme for eukaryotes. Additionally, we investigated the dependence of decay rates on sequence composition, that is, the presence or absence of short mRNA motifs in various regions of the mRNA transcript. Our analysis recovers the positive correlation of mRNA decay with known AU-rich mRNA motifs, but we also uncover further short mRNA motifs that show statistically significant correlation with decay. However, we also note that none of these motifs are strong predictors of mRNA decay rate, indicating that the regulation of mRNA decay is more complex and may involve the cooperative binding of several RNA-binding proteins at different sites. PMID- 12902381 TI - A population threshold for functional polymorphisms. AB - We sequenced 114 genes (for DNA repair, cell cycle arrest, apoptosis, and detoxification)in a mixed human population and observed a sudden increase in the number of functional polymorphisms below a minor allele frequency of approximately 6%. Functionality is assessed by considering the ratio in the number of nonsynonymous single nucletide polymorphisms (SNPs)to the number of synonymous or intron SNPs. This ratio is steady from below 1% in frequency-that regime traditionally associated with rare Mendelian diseases-all the way up to about 6% in frequency, after which it falls precipitously. We consider possible explanations for this threshold effect. There are four candidates as follows: (1). deleterious variants that have yet to be purified from the population, (2). balancing selection, in which a selective advantage accrues to the heterozygotes, (3). population-specific functional polymorphisms, and (4). adaptive variants that are accumulating in the population as a response to the dramatic environmental changes of the last 7000 approximately 17000 years. PMID- 12902382 TI - Effects of recombination rate and gene density on transposable element distributions in Arabidopsis thaliana. AB - Transposable elements (TEs) comprise a major component of eukaryotic genomes, and exhibit striking deviations from random distribution across the genomes studied, including humans, flies, nematodes, and plants. Although considerable progress has been made in documenting these patterns, the causes are subject to debate. Here, we use the genome sequence of Arabidopsis thaliana to test for the importance of competing models of natural selection against TE insertions. We show that, despite TE accumulation near the centromeres, recombination does not generally correlate with TE abundance, suggesting that selection against ectopic recombination does not influence TE distribution in A. thaliana. In contrast, a consistent negative correlation between gene density and TE abundance, and a strong under-representation of TE insertions in introns suggest that selection against TE disruption of gene expression is playing a more important role in A. thaliana. High rates of self-fertilization may reduce the importance of recombination rate in genome structuring in inbreeding organisms such as A. thaliana and Caenorhabditis elegans. PMID- 12902383 TI - Estimating the repeat structure and length of DNA sequences using L-tuples. AB - In shotgun sequencing projects, the genome or BAC length is not always known. We approach estimating genome length by first estimating the repeat structure of the genome or BAC, sometimes of interest in its own right, on the basis of a set of random reads from a genome project. Moreover, we can find the consensus for repeat families before assembly. Our methods are based on the l-tuple content of the reads. PMID- 12902384 TI - Detection and assignment of mutations and minihaplotypes in human DNA using peptide mass signature genotyping (PMSG): application to the human RDS/peripherin gene. AB - Peptide mass-signature genotyping (PMSG) is a scanning genotyping method that identifies mutations and polymorphisms by translating the sequence of interest in more than one reading frame and measuring the masses of the resulting peptides by mass spectrometry. PMSG was applied to the RDS/peripherin gene of 16 individuals from a family exhibiting autosomal dominant macular degeneration. The method revealed an A-->T transversion in the 5' splice site of intron 2 that is the likely cause of the disease. It also revealed four different minihaplotypes in exon 3 that represent particular combinations of SNPs at four different locations. This study demonstrates the utility of PMSG for identifying and characterizing point mutations and local minihaplotypes that are not readily analyzed by other approaches. PMID- 12902385 TI - Automated detection of informative combined effects in genetic association studies of complex traits. AB - There is a growing body of evidence suggesting that the relationships between gene variability and common disease are more complex than initially thought and require the exploration of the whole polymorphism of candidate genes as well as several genes belonging to biological pathways. When the number of polymorphisms is relatively large and the structure of the relationships among them complex, the use of data mining tools to extract the relevant information is a necessity. Here, we propose an automated method for the detection of informative combined effects (DICE) among several polymorphisms (and nongenetic covariates) within the framework of association studies. The algorithm combines the advantages of the regressive approaches with those of data exploration tools. Importantly, DICE considers the problem of interaction between polymorphisms as an effect of interest and not as a nuisance effect. We illustrate the method with three applications on the relationship between (1). the P-selectin gene and myocardial infarction, (2). the cholesteryl ester transfer protein gene and plasma high density-lipoprotein cholesterol concentration, and (3). genes of the renin angiotensin-aldosterone system and myocardial infarction. The applications demonstrated that the method was able to recover results already found using other approaches, but in addition detected biologically sensible effects not previously described. PMID- 12902386 TI - Expanding the use of zymography by the chemical linkage of small, defined substrates to the gel matrix. AB - In the postgenomic era, the comprehensive proteomic analysis of metabolic and signaling pathways is inevitably faced with the challenge of large-scale identification and characterization of polypeptides with a particular enzymatic activity. Previous work has shown that a wide variety of enzymatic activities of microbial, plant, and animal origin can be assigned to individual polypeptides using in-gel activity staining (zymography). However, a number of limitations, such as special substrate requirements, the lack of a standard procedure, and difficulties in distinguishing enzymes with overlapping activities have precluded the widespread use of zymography as a routine laboratory method. Here we demonstrate that, by employing small-defined substrates that are covalently attached to the gel matrix, we can largely overcome the aforementioned problems and assay readily a number of different classes of enzymatic activities within gels after standard SDS-polyacrylamide electrophoresis. Moreover, this development is compatible with the two-dimensional separation of proteins and thus has great potential in the high-throughput screening and characterization of complex biological and clinical samples. PMID- 12902387 TI - A cattle-human comparative map built with cattle BAC-ends and human genome sequence. AB - As a step toward the goal of adding the cattle genome to those available for multispecies comparative genome analysis, 40,224 cattle BAC clones were end sequenced, yielding 60,547 sequences (BAC end sequences, BESs) after trimming with an average read length of 515 bp. Cattle BACs were anchored to the human and mouse genome sequences by BLASTN search, revealing 29.4% and 10.1% significant hits (E < e-5), respectively. More than 60% of all cattle BES hits in both the human and mouse genomes are located within known genes. In order to confirm in silico predictions of orthologyand their relative position on cattle chromosomes, 84 cattle BESs with similarity to sequences on HSA11 were mapped using a cattle hamster radiation hybrid (RH) panel. Resulting RH maps of BTA15 and BTA29 cover approximately 85% of HSA11 sequence, revealing a complex patchwork shuffling of segments not explained by a simple translocation followed by internal rearrangements. Overlay of the mouse conserved syntenies onto HSA11 revealed that segmental boundaries appear to be conserved in all three species. The BAC clone based comparative map provides a foundation for the evolutionary analysis of mammalian karyotypes and for sequencing of the cattle genome. PMID- 12902388 TI - Dlx2 progenitor migration in wild type and Nkx2.1 mutant telencephalon. AB - The transcription factor Dlx2 is expressed widely throughout the ventral telencephalon. We have examined the in vitro and in vivo migration of Dlx2 progenitors originating from the different ganglionic eminences of both wild type and Nkx2.1 mutant animals. By examining the expression of tauLacZ targeted into the Dlx2 locus we were able to visualize the distribution of cells expressing this gene at both embryonic and postnatal stages. This analysis suggested that Dlx2-expressing cells traverse a number of characteristic migratory routes to populate both cortical and subcortical regions. We also examined how these patterns of migration were affected in Nkx2.1 mutant animals. In these mutants, the early but not late populations of Dlx2-expressing cells originating in the ventral telencephalon that migrate to the cortex are lost. This recovery may be, at least in part, a result of the late migration of Dlx2 progenitors from the caudal ganglionic eminences (CGE), which, based on our previous work, does not appear to require Nkx2.1 gene function. PMID- 12902389 TI - Regional brain activation associated with different performance patterns during learning of a complex motor skill. AB - In understanding the brain's response to extensive practice and development of high-level, expert skill, a key question is whether the same brain structures remain involved throughout the different stages of learning and a form of adaptation occurs, or a new functional circuit is formed with some structures dropping off and others joining. After training subjects on a set of complex motor tasks (tying knots), we utilized fMRI to observe that in subjects who learned the task well new regional activity emerged in posterior medial structures, i.e. the posterior cingulate gyrus. Activation associated with weak learning of the knots involved areas that mediate visual spatial computations. Brain activity associated with no substantive learning indicated involvement of areas dedicated to the declarative aspects learning such as the anterior cingulate and prefrontal cortex. The new activation for the pattern of strong learning has alternate interpretations involving either retrieval during episodic memory or a shift toward non-executive cognitive control of the task. While these interpretations are not resolved, the study makes clear that single time-point images of motor skill can be misleading because the brain structures that implement action can change following practice. PMID- 12902390 TI - Representation of the perceived 3-D object shape in the human lateral occipital complex. AB - We used human functional magnetic resonance imaging (fMRI) to test whether the human lateral occipital complex (LOC), an area known to be involved in the analysis of visual shape, represents the perceived 3-D shape of objects or simply their 2-D contours. We employed an fMRI adaptation paradigm, in which repeated presentation of a stimulus results in decreased responses compared to responses to different stimuli. We found adaptation in the LOC for images of objects with the same perceived 3-D shape structure but different 2-D contours that resulted from small rotations of the objects in the frontal plane or in depth. However, no adaptation was observed in the LOC for images of objects that had the same 2-D contours but differed in their perceived 3-D shape; namely, 2-D silhouettes versus 3-D shaded images of objects, or convex versus concave objects. Differences in the fMRI adaptation responses across subregions in the LOC suggest that different neural populations in the LOC may mediate different mechanisms for the processing of object features. PMID- 12902392 TI - Blockade of GABA(B) receptors alters the tangential migration of cortical neurons. AB - To better understand the role of neurotransmitter receptors in neuronal differentiation and maturation a detailed knowledge of their identity, location and function in the plasma membrane of specific neuronal populations during development is required. Combining pre-embedding immunocytochemistry with cell tracking in embryonic brain slice cultures we show that virtually all neurons (approximately 98%) migrating through the lower intermediate zone (LIZ) on their way from the medial ganglionic eminence to the cerebral cortex, express GABA(B)R1. Blockade of GABA(B)Rs with a specific antagonist, CGP52432, resulted in a concentration-dependent accumulation of these tangentially migrating neurons in the ventricular/sub-ventricular zones (VZ/SVZ) of the cortex and fewer cells were observed in the cortical plate/marginal zone (CP/MZ) and LIZ. Moreover, they had significantly shorter leading processes compared with similar migrating cells in control slices. Electrophysiological recording in LIZ and CP cells revealed no direct effect of either CGP52432 or the GABA(B)R agonist, baclofen, on resting membrane properties suggesting that the effect of CGP52432 on migration might be mediated through a metabotropic action or the regulation of release of factors controlling migration. These results suggest that GABA(B)Rs have an important modulatory role in the migration of cortical interneurons. PMID- 12902393 TI - Cerebellar volume of musicians. AB - There is evidence that the cerebellum is involved in motor learning and cognitive function in humans. Animal experiments have found structural changes in the cerebellum in response to long-term motor skill activity. We investigated whether professional keyboard players, who learn specialized motor skills early in life and practice them intensely throughout life, have larger cerebellar volumes than matched non-musicians by analyzing high-resolution T(1)-weighted MR images from a large prospectively acquired database (n = 120). Significantly greater absolute (P = 0.018) and relative (P = 0.006) cerebellar volume but not total brain volume was found in male musicians compared to male non-musicians. Lifelong intensity of practice correlated with relative cerebellar volume in the male musician group (r = 0.595, P = 0.001). In the female group, there was no significant difference noted in volume measurements between musicians and non-musicians. The significant main effect for gender on relative cerebellar volume (F = 10.41, P < 0.01), with females having a larger relative cerebellar volume, may mask the effect of musicianship in the female group. We propose that the significantly greater cerebellar volume in male musicians and the positive correlation between relative cerebellar volume and lifelong intensity of practice represents structural adaptation to long-term motor and cognitive functional demands in the human cerebellum. PMID- 12902391 TI - Expression of the transcription factor, tailless, is required for formation of superficial cortical layers. AB - The gene tailless (tlx) encodes a forebrain-restricted transcription factor that is robustly expressed in progenitor cells of the ventricular and subventricular zones during neurogenesis. To investigate the role of tlx in neocortical development we generated a targeted deletion of tlx by homologous recombination. Here we compared the lamination, connectivity and patterning of cortical regions in adult tlx-/- mice and their wild-type littermates. We found first that neocortical thickness is reduced by 20% in mutant animals; most of this reduction is due to a diminution of supragranular layers, while layer I and layers IV through VI are relatively intact cytoarchitecturally. Consistent with this, the cross-sectional area of the corpus callosum is reduced by over 40%. Second, thalamocortical and intrinsic excitatory circuits in tlx-/- mice exhibit an essentially normal distribution from layer IV to the white matter, but are reduced superficial to layer IV. Finally, within parietal cortex of mutant mice a vibrissa-like pattern of cortical barrels is present in the expected rostro caudal location. These observations indicate that loss of tlx function most severely affects generation and differentiation of neurons destined for superficial cortical layers. Thus, tlx may be important in sustaining the progenitor cell population throughout late prenatal development. Establishment of functional cortical areas, and development of basic patterns of thalamocortical and intra-cortical circuits occurs independently of tlx function. PMID- 12902394 TI - Age-related dendritic and spine changes in corticocortically projecting neurons in macaque monkeys. AB - Alterations in neuronal morphology occur in primate cerebral cortex during normal aging, vary depending on the neuronal type, region and cortical layer, and have been related to memory and cognitive impairment. We analyzed how such changes affect a specific subpopulation of cortical neurons forming long corticocortical projections from the superior temporal cortex to prefrontal area 46. These neurons were identified by retrograde transport in young and old macaque monkeys. Dendritic arbors of retrogradely labeled neurons were visualized in brain slices by intracellular injection of Lucifer Yellow, and reconstructed three dimensionally using computer-assisted morphometry. Total dendritic length, numbers of segments, numbers of spines, and spine density were analyzed in layer III pyramidal neurons forming the projection considered. Sholl analysis was used to determine potential age-related changes in dendritic complexity. We observed statistically significant age-related decreases in spine numbers and density on both apical and basal dendritic arbors in these projection neurons. On apical dendrites, changes in spine numbers occurred mainly on the proximal dendrites but spine density decreased uniformly among the different branch orders. On basal dendrites, spine numbers and density decreased preferentially on distal branches. Regressive dendritic changes were observed only in one particular portion of the apical dendrites, with the general dendritic morphology and extent otherwise unaffected by aging. In view of the fact that there is no neuronal loss in neocortex and hippocampus in old macaque monkeys, it is possible that the memory and cognitive decline known to occur in these animals is related to rather subtle changes in the morphological and molecular integrity of neurons subserving identifiable neocortical association circuits that play a critical role in cognition. PMID- 12902395 TI - The role of parietal cortex in awareness of self-generated movements: a transcranial magnetic stimulation study. AB - Awareness of self-generated movements arises from comparing motor plans, and the accompanying (hypothetical) efference copy, with the visual and proprioceptive consequences of movement. Here we used repetitive transcranial magnetic stimulation (rTMS) to investigate the role of a posterior region in the superior parietal lobule (SPL) in this process. Nine healthy volunteers performed a finger extension actively and passively while wearing a CyberGlove; the glove recorded these (actual) finger movements and used this information in real time to move a virtual hand displayed on a computer screen. To assess the participant's awareness of movement onset, we introduced a delay between the onset of the actual and virtual movement (60-270 ms, 30 ms increments); the task was to judge whether the virtual hand movements were delayed relative to the actual hand movements. Low-frequency rTMS (15 min, 0.6 Hz) was applied either over the left SPL or the left temporal cortex (control site) to decrease excitability of these regions and, in turn, test their role in the awareness of self-generated movement. Following the SPL stimulation, participants' assessments of asynchrony were impaired for active but not passive movements. No significant changes were observed after rTMS applied over the control site. We suggest that these findings are consistent with the role of the SPL in evaluating the temporal congruency of peripheral (visual) and central (efference copy) signals associated with self generated movements. As such, this region may contribute to the sense of 'agency' and its disturbances in disorders such as apraxia and schizophrenia. PMID- 12902396 TI - NMDA receptors in the rat orbital prefrontal cortex are involved in guidance of instrumental behaviour under reversal conditions. AB - The orbital prefrontal cortex (OPFC) might be particularly involved in adapting instrumental responses to changes of stimulus-reward contingencies. We investigated whether signals in the OPFC transmitted via N-methyl-D-aspartate (NMDA) receptors are critical for learning a reversal of stimulus-reward contingencies. Rats were trained in a reaction time (RT) task demanding conditioned lever release with discriminative stimuli signalling in advance the upcoming reward magnitude (one or five pellets). After acquisition, RT of responses with expectancy of high reward magnitude were significantly shorter. Thereafter, stimulus-reward contingencies were reversed and rats received intra OPFC infusions of saline or of the NMDA receptor antagonist AP5. Reversal learning was tested for 6 days, with microinfusions being given on days 1, 3 and 5. Intra-OPFC blockade of NMDA receptors impaired the learning of a reversal of previously acquired stimulus-reward magnitude contingencies: (i) latencies of correct responses were generally shortened, regardless of the response-associated reward magnitude; (ii) the proportion of premature responses was increased; and (iii) responses were not guided by the current significance of the reward predicting stimuli. These findings provide novel evidence for NMDA-receptor dependent plasticity in the OPFC in reversal learning. PMID- 12902397 TI - Activation of the supplementary motor area (SMA) during performance of visually guided movements. AB - The supplementary motor area (SMA) has long been thought to have a special role in the internal generation of complex movements. Yet, a number of recent functional imaging studies indicate that the SMA is activated during the execution of simple movements guided by sensory cues. The extent of participation of the cingulate motor areas in visually guided movements also is unclear. To explore these issues we used the 2-deoxyglucose (2DG) technique to measure functional activation in the motor areas on the medial wall of the hemisphere in monkeys trained to perform visually guided reaching movements to randomly presented targets. This approach enabled us to make precise comparisons between sites of activation and the location of specific premotor areas on the medial wall of the hemisphere. We found that the SMA was strongly activated during reaching to different visual targets. Indeed, its activation was comparable to that of the primary motor cortex (M1). In contrast, none of the cingulate motor areas displayed significantly increased activation specifically related to arm movements. Our results provide further support for the involvement of the SMA in visually guided movements. Furthermore, our observations suggest that during externally guided reaching, SMA activation is tightly coupled to that of M1, but dissociated from that of the cingulate motor areas. PMID- 12902398 TI - Evidence for a rostral-to-caudal somatotopic organization in human primary somatosensory cortex with mirror-reversal in areas 3b and 1. AB - Medial-to-lateral somatotopy is a well-established feature of the human primary somatosensory cortex (SI); however, it is unknown whether, similarly to non-human primates, a rostral-to-caudal somatotopic arrangement exists as well. Therefore, in this functional magnetic resonance imaging (fMRI) study on eight healthy human subjects, five circumscribed skin areas sequentially located on the third finger and the palm of the hand were stimulated with innocuous electrical pulses. Within area 3b of contralateral SI, successive cortical representation sites ordered in a rostral-to-caudal fashion were seen in the group analysis and in six individual subjects. The fingertip was located most rostrally, whereas the proximal parts of the finger as well as the distal palm were represented at more caudal locations. Within area 1, the group analysis revealed a similar pattern of discrete representations. However, in contrast to area 3b, the fingertip was located most caudally, whereas the more proximal parts of the finger were found to be represented rostrally within area 1. Thus, the representation pattern of area 1 appeared as a 'mirror image' of that of area 3b. In comparison to the representations of the finger and the distal palm, the proximal palm was found to be represented at a more medial position of the postcentral gyrus. PMID- 12902399 TI - Dominance for vestibular cortical function in the non-dominant hemisphere. AB - The aim of this (15)O-labelled H(2)O bolus positron emission tomography (PET) study was to analyse the hemispheric dominance of the vestibular cortical system. Therefore, the differential effects of caloric vestibular stimulation (right or left ear irrigation with warm water at 44 degrees C) on cortical and subcortical activation were studied in 12 right-handed and 12 left-handed healthy volunteers. Caloric irrigation induces a direction-specific sensation of rotation and nystagmus. Significant regional cerebral blood flow increases were found in a network within both hemispheres, including the superior frontal gyrus/sulcus, the precentral gyrus and the inferior parietal lobule with the supramarginal gyrus. These areas correspond best to the cortical ocular motor centres, namely the prefrontal cortex, the frontal eye field and the parietal eye field, known to be involved in the processing of caloric nystagmus. Furthermore, distinct temporo parietal activations could be separated in the posterior part of the insula with the adjacent superior temporal gyrus, the inferior parietal lobule and precuneus. These areas fit best to the human homologues of multisensory vestibular cortex areas identified in the monkey and correspond to the parieto-insular vestibular cortex (PIVC), the visual temporal sylvian area (VTS) and areas 7 and 6. Further cortical activations were seen in the anterior insula, the inferior frontal gyrus and anterior cingulum. The subcortical activation pattern in the putamen, thalamus and midbrain is consistent with the organization of efferent ocular motor pathways. Cortical and subcortical activation of the described areas was bilateral during monaural stimulation, but predominant in the hemisphere ipsilateral to the stimulated ear and exhibited a significant right hemispheric dominance for vestibular and ocular motor structures in right-handed volunteers. Similarly, a significant left hemispheric dominance was found in the 12 left handed volunteers. Thus, this PET study showed for the first time that cortical and subcortical activation by vestibular caloric stimulation depends (i) on the handedness of the subjects and (ii) on the side of the stimulated ear. Maximum activation was therefore found when the non-dominant hemisphere was ipsilateral to the stimulated ear, i.e. in the right hemisphere of right-handed subjects during caloric irrigation of the right ear and in the left hemisphere of left handed subjects during caloric irrigation of the left ear. The localization of handedness and vestibular dominance in opposite hemispheres might conceivably indicate that the vestibular system and its hemispheric dominance, which matures earlier during ontogenesis, determine right- or left-handedness. PMID- 12902400 TI - Plasmodium falciparum apical membrane antigen 1 (PfAMA-1) is translocated within micronemes along subpellicular microtubules during merozoite development. AB - During the assembly of Plasmodium falciparum merozoites within the schizont stage, the parasite synthesizes and positions three sets of secretory vesicles (rhoptries, micronemes and dense granules) that are active during red cell invasion. There are up to 40 micronemes per merozoite, shaped like long-necked bottles, about 160 nm long and 65 nm at their widest diameter. On their external surfaces, they bear bristle-like filaments, each 3-4 nm thick and 25 nm long. Micronemes are translocated from a single Golgi-like cisterna near the nucleus along a band of two or three subpellicular microtubules to the merozoite apex, where they dock with the rhoptry tips. Dense granules are also formed around the periphery of the Golgi cisternae but their distribution is unrelated to microtubules. Three polyclonal antibodies raised against the recombinant PfAMA-1 ectodomain sequence recognizing both the 83 kDa and processed 66 kDa molecules label the peripheries of translocating and mature micronemes but do not label rhoptries significantly at any stage of merozoite development within schizonts. This result confirms that PfAMA-1 is a micronemal protein, and indicates that within the microneme it is located near or inserted into this organelle's boundary membrane. PMID- 12902401 TI - Mechanisms in LPA-induced tumor cell migration: critical role of phosphorylated ERK. AB - Lysophosphatidic acid (LPA) is a serum-borne phospholipid with hormone and growth factor-like properties. LPA has been shown to modulate tumor cell invasion and malignant cell growth. Here, we report that two human pancreatic carcinoma cell lines, PANC-1 and BxPC-3, express functionally active LPA receptors coupled to pertussis toxin-sensitive Gi/o-proteins. In contrast to other cell types, LPA does not act as a mitogen, but is an efficacious stimulator of cell migration of these tumor cells. LPA-induced chemotaxis is markedly dependent on activation of PTX-sensitive heterotrimeric G-proteins, on activation of the small GTPases Ras, Rac and RhoA, and on GTPase-dependent activation of ERK. LPA-induced ERK activation results in a transient translocation of the phosphorylated ERK to newly forming focal contact sites at the leading edge of the migrating cells. Inhibition of ERK activation and its subsequent translocation impaired LPA induced chemotaxis and LPA-induced actin reorganization. Thus, pancreatic tumor cell migration in response to LPA is essentially controlled by activation of a Gi/o-ERK pathway and requires the LPA-induced activation of Ras, Rac1 and RhoA. PMID- 12902402 TI - Endothelial expression of the alpha6beta4 integrin is negatively regulated during angiogenesis. AB - Development and homeostasis of the vascular system requires integrin-facilitated cellular adhesion, migration, proliferation and survival. A specific role for the alpha6beta4 integrin in the vasculature, however, has not been identified. Using immunohistochemistry, we observed alpha6beta4 expression on the dermal microvasculature of human foreskin. Analysis of individual cells isolated from trypsin-disrupted foreskin tissue indicated that alpha6beta4 was expressed by a subset of epithelial and endothelial cells, and not by smooth muscle cells. Expression of alpha6beta4 was also analyzed during new vessel growth using explants of human saphenous vein cultured in fibrinogen gels. The results indicate that alpha6beta4 is not expressed by outgrowing endothelial cells, and is downregulated by the original alpha6beta4-positive endothelial cells of the explant. To determine whether alpha6beta4 is expressed during angiogenesis in vivo, the expression of the beta4 subunit was analyzed during the development of the mouse mystacial (whisker) pad. Immunohistochemical staining of the whisker pad indicates that beta4 is expressed by the adult vasculature. To identify when and where beta4 is turned on in the vasculature, we examined the whisker pads from the developing embryo (E19.5 pc), and from postnatal days zero (P0), three (P3) and seven (P7) pups. The expression of alpha6beta4 was found to be turned on spatially and temporally from caudal to rostral regions and from the deep to superficial vasculature, correlating with the maturation of the whisker pad and its corresponding vasculature. Together, these findings suggest a potential role for alpha6beta4 as a negative component of the angiogenic switch, whereas expression of alpha6beta4 on the adult vasculature may indicate regions requiring additional adhesive mechanisms. PMID- 12902403 TI - Barrier-to-autointegration factor plays crucial roles in cell cycle progression and nuclear organization in Drosophila. AB - Barrier-to-autointegration factor (BAF) is potentially a DNA-bridging protein, which directly associates with inner nuclear membrane proteins carrying LEM domains. These features point to a key role in regulation of nuclear function and organization, dependent on interactions between the nuclear envelope and chromatin. To understand the functions of BAF in vivo, Drosophila baf null mutants generated by P-element-mediated imprecise excision were analyzed. Homozygous null mutants showed a typical mitotic mutant phenotype: lethality at the larval-pupal transition with small brains and missing imaginal discs. Mitotic figures were decreased but a defined anaphase defect as reported for C. elegans RNAi experiments was not observed in these small brains, suggesting a different phase or phases of cell cycle arrest. Specific abnormalities in interphase nuclear structure were frequently found upon electron microscopic examination of baf null mutants, with partial clumping of chromatin and convolution of nuclear shape. At the light microscopic level, grossly aberrant nuclear lamina structure and B-type lamin distribution correlated well with the loss of detectable amounts of BAF protein from nuclei. Together, these data represent evidence of BAF's anticipated function in mediating interactions between the nuclear envelope and interphase chromosomes. We thus conclude that BAF plays essential roles in nuclear organization and that these BAF functions are required in both M phase and interphase of the cell cycle. PMID- 12902404 TI - Loss of responsiveness to chemotactic factors by deletion of the C-terminal protein interaction site of angiomotin. AB - We have recently identified a novel protein, named angiomotin, by its ability to bind the angiogenesis inhibitor angiostatin in the yeast two-hybrid system. Angiomotin belongs to a family with two other members, AmotL-1 and -2 characterized by coiled-coil and C-terminal PDZ binding domains. Here we show that the putative PDZ binding motif of angiomotin serves as a protein recognition site and that deletion of three amino acids in this site results in inhibition of chemotaxis. Furthermore, endothelial cells expressing mutant angiomotin failed to migrate and form tubes in an in vitro tube formation assay. To study the effect of angiomotin on embryonic angiogenesis, we generated transgenic mice expressing wild-type angiomotin and the C-terminal deletion mutant driven by the endothelial cell-specific receptor tyrosine kinase (TIE) promoter. Expression of mutant angiomotin in endothelial cells inhibited migration into the neuroectoderm and intersomitic regions resulting in death at embryonic day 9.5. In contrast, mice expressing wild-type angiomotin developed normally and were fertile. These results suggest that the putative PDZ binding motif of angiomotin plays a critical role in regulating the responsiveness of endothelial cells to chemotactic cues. PMID- 12902405 TI - Chlamydia trachomatis uses host cell dynein to traffic to the microtubule organizing center in a p50 dynamitin-independent process. AB - Chlamydiae are pathogenic obligate intracellular bacteria with a biphasic developmental cycle that involves cell types adapted for extracellular survival (elementary bodies, EBs) and intracellular multiplication (reticulate bodies, RBs). The intracellular development of chlamydiae occurs entirely within a membrane-bound vacuole termed an inclusion. Within 2 hours after entry into host cells, Chlamydia trachomatis EBs are trafficked to the perinuclear region of the host cell and remain in close proximity to the Golgi apparatus, where they begin to fuse with a subset of host vesicles containing sphingomyelin. Here, we provide evidence that chlamydial migration from the cell periphery to the peri-Golgi region resembles host cell vesicular trafficking. Chlamydiae move towards the minus end of microtubules and aggregate at the microtubule-organizing center (MTOC). In mammalian cells the most important minus-end-directed microtubule motor is cytoplasmic dynein. Microinjection of antibodies to a subunit of cytoplasmic dynein inhibited movement of chlamydiae to the MTOC, whereas microinjection of antibodies to the plus-directed microtubule motor, kinesin, had no effect. Surprisingly, overexpression of the protein p50 dynamitin, a subunit of the dynactin complex that links vesicular cargo to the dynein motor in minus directed vesicle trafficking, did not abrogate chlamydial migration even though host vesicle transport was inhibited. Nascent chlamydial inclusions did, however, colocalize with the p150(Glued) dynactin subunit, which suggests that p150(Glued) may be required for dynein activation or processivity but that the cargo-binding activity of dynactin, supplied by p50 dynamitin subunits and possibly other subunits, is not. Because chlamydial transcription and translation were required for this intracellular trafficking, chlamydial proteins modifying the cytoplasmic face of the inclusion membrane are probable candidates for proteins fulfilling this function. PMID- 12902406 TI - Suprabasal alpha6beta4 integrin expression in epidermis results in enhanced tumourigenesis and disruption of TGFbeta signalling. AB - Inappropriate alpha6beta4 integrin expression correlates with a high risk of tumour progression in stratified squamous epithelia. Targeted expression of alpha6beta4 in the suprabasal layers of transgenic mouse epidermis dramatically increased the frequency of papillomas, carcinomas and metastases induced by chemical carcinogenesis, independent of the beta4 cytoplasmic domain. Suprabasal alpha6beta4 also perturbed transforming growth factor beta (TGFbeta) signalling as demonstrated by decreased nuclear Smad2 in transgenic epidermis and tumours. In cultured keratinocytes, suprabasal alpha6beta4 relieved TGFbeta-mediated growth inhibition and blocked nuclear translocation of activated Smad2/3. Responsiveness to TGFbeta could be restored by inhibiting cadherin-mediated cell cell adhesion or phosphoinositide 3-kinase (PI3-K) activity, but not by inhibiting mitogen-activated protein kinase (MAPK) activity. These data suggest that suprabasal alpha6beta4 promotes tumourigenesis by preventing TGFbeta from suppressing clonal expansion of initiated cells in the epidermal basal layer. PMID- 12902407 TI - Who should read and interpret 18F-FDG PET studies? PMID- 12902408 TI - Clinical performance of PET/CT in evaluation of cancer: additional value for diagnostic imaging and patient management. AB - This study assessed the clinical performance of a combined PET/CT system using (18)F-FDG in oncologic patients. METHODS: (18)F-FDG PET/CT was used to evaluate 204 patients with 586 suspicious lesions. All patients had available follow-up data, enabling assessment of the clinical significance of hybrid PET/CT findings. Differences in interpretation between PET, CT, and fused PET/CT data were prospectively documented for detection, localization, and characterization of each evaluated site. The additional value of PET/CT for data interpretation over that of separate PET and CT was classified into several criteria, including change in lesion characterization to either definitely benign or definitely malignant, precise anatomic localization of malignant (18)F-FDG uptake, and retrospective lesion detection on PET and CT. The clinical impact of information provided by PET/CT on patient management was assessed on the basis of follow-up data concerning further diagnostic or therapeutic approach. Analysis of data was performed for the whole study population, for different types of cancer, and for different anatomic sites. RESULTS: PET/CT provided additional information over the separate interpretation of PET and CT in 99 patients (49%) with 178 sites (30%). PET/CT improved characterization of equivocal lesions as definitely benign in 10% of sites and as definitely malignant in 5% of sites. It precisely defined the anatomic location of malignant (18)F-FDG uptake in 6%, and it led to retrospective lesion detection on PET or CT in 8%. The results of PET/CT had an impact on the management of 28 patients (14%). Hybrid PET/CT data obviated the need for further evaluation in 5 patients, guided further diagnostic procedures in 7 patients, and assisted in planning therapy for 16 patients. CONCLUSION: Hybrid PET/CT improves the diagnostic interpretation of (18)F-FDG PET and CT in cancer patients and has an impact on both diagnostic and therapeutic aspects of patient management. PMID- 12902409 TI - Measurement of the extracellular space in brain tumors using 76Br-bromide and PET. AB - Brain edema significantly contributes to the clinical course of human brain tumor patients. There is evidence that an enlargement of the extracellular space (ECS) is involved in the development of brain edema. Although T2-weighted magnetic resonance (T2-MR) images represent brain edema by its increased water content, they do not differentiate ECS enlargement from increased intracellular water content. METHODS: On the basis of the known distribution of bromide in the ECS, we used (76)Br-bromide and PET to measure the regional ECS in 9 brain tumor patients. Transport rate constants and the distribution volume (DV) of (76)Br bromide in normal brain and tumor were derived from dynamic PET scans and the measured (76)Br-bromide concentration in arterial plasma. We evaluated different models regarding their reliability in estimating the ECS. RESULTS: Assuming that the DV of (76)Br-bromide represents the ECS, robust estimates were possible for all investigated regions. In normal brain, ECS was within a narrow range-for example, occipital lobe, 19.9% +/- 3.1%-and was lower in 2 dexamethasone-treated patients compared with untreated patients. In 7 of 9 tumors, increased ECS ranged between 43.8% and 61.1%. ECS increases were confined to the tumor mass and did not extend into peritumoral edematous brain. Two patients with large hyperintense lesions according to T2-MR images showed normal ECS values within the lesion. CONCLUSION: (76)Br-Bromide PET allows a quantitative measurement of the ECS in brain edema and in normal brain. The discrepancies between lesions shown by T2 MRI and regional ECS enlargement as measured with PET challenge the concept of tumor-induced brain edema. PMID- 12902410 TI - Mediastinal lymph node involvement in non-small cell lung cancer: evaluation with 99mTc-tetrofosmin SPECT and comparison with CT. AB - In patients with non-small cell lung cancer (NSCLC), surgical resection offers the best chance of cure. The preoperative assessment of mediastinal lymph node involvement is crucial to selecting those patients for whom surgery is indicated. METHODS: To evaluate the possible clinical role of (99m)Tc-tetrofosmin scintigraphy in the presurgical detection of mediastinal node metastases from NSCLC, we performed a prospective comparative study with CT on 83 patients (48 men, 35 women; age range, 38-81 y) with primary NSCLC (36 adenocarcinomas, 39 epidermoid squamous cell carcinomas, and 8 large cell anaplastic carcinomas). They underwent chest SPECT 20 min after (99m)Tc-tetrofosmin injection (740 MBq intravenously). The metastatic involvement of mediastinal nodes was assessed by histologic examination after mediastinoscopy or thoracotomy. Both chest CT and (99m)Tc-tetrofosmin scintigraphy were performed within 2 wk before the surgical staging. RESULTS: Metastatic mediastinal lymph nodes were found in 35 patients. (99m)Tc-Tetrofosmin imaging in assessing the mediastinal involvement yielded a sensitivity of 85.7%, a specificity of 89.6%, and an accuracy of 88.0%; CT results were 68.6%, 75.0%, and 72.3%, respectively. SPECT accuracy was significantly higher than CT accuracy (P < 0.05). However, precise anatomic localization of (99m)Tc-tetrofosmin uptake in the mediastinum was not always present on SPECT images. (99m)Tc-Tetrofosmin SPECT precisely detected the presence or absence of lymph node metastases in 33 of the 36 patients with positive CT findings (enlarged mediastinal nodes with a short axis > or =1 cm), with an accuracy (91.7%) significantly higher (P < 0.05) than that of CT (66.7%). CONCLUSION: (99m)Tc-Tetrofosmin SPECT is a useful presurgical noninvasive method to assess mediastinal lymph node involvement in NSCLC. In particular, it could play a clinical role in reducing the number of invasive staging surgical procedures in selected patients, especially in those with enlarged lymph nodes at CT. Fusing SPECT with CT images could further improve the interpretation of the scintigraphic data. PMID- 12902411 TI - Prognostic value of PET using 18F-FDG in Hodgkin's disease for posttreatment evaluation. AB - Detection of relapse after completion of therapy in patients with Hodgkin's disease (HD) and non-Hodgkin's lymphomas (NHL) constitutes an important challenge in modern medical imaging. An accurate assessment of the presence of residual disease is essential to determine which patients would benefit from additional therapy. The objective of this study was to assess the diagnostic accuracy of (18)F-FDG PET in detecting residual disease or relapse during the posttherapy period in patients with HD in comparison with CT. We also established different predictive values for (18)F-FDG PET according to the time interval between the end of therapy and the PET study. METHODS: Forty-eight patients with HD underwent (18)F-FDG PET after the completion of chemotherapy (median, 58 d) between March 1999 and April 2002. Disease-free intervals and proportions were calculated using the Kaplan-Meier method. Standardized uptake values of the most active lesion in each patient with a positive study were also measured. PET and CT results were compared with clinical follow-up, with relapse being defined by a positive biopsy or the introduction of a second-line treatment. RESULTS: Thirty-four patients were still disease-free during a mean follow-up of 605 d. Fourteen patients relapsed during a mean follow-up of 197 d. The sensitivity and specificity of (18)F-FDG PET to predict relapse were 79% and 97%, respectively. The positive predictive value and the negative predictive value were both equal to 92%. The diagnostic accuracy of (18)F-FDG PET (92%) was significantly higher than the accuracy of CT (56%) (P < 0.0005). Patients with positive (18)F-FDG PET also had a far shorter median disease-free interval (79 d) than those with positive CT (disease-free proportion of 52% at 1,143 d) (P = 0.0046). The 3 cases of false negative (18)F-FDG PET studies that we observed occurred in patients who underwent their PET study within the first 49 d after the end of chemotherapy. CONCLUSION: Positive (18)F-FDG PET after the end of therapy in HD patients is a strong predictor of relapse. A negative PET study is also an excellent predictor of good prognosis. The diagnostic accuracy of (18)F-FDG PET to assess the presence of residual disease after therapy is superior to that of CT. PMID- 12902412 TI - Nonvisualization of axillary sentinel node during lymphoscintigraphy: is there a pathologic significance in breast cancer? AB - The aim of this study was to define the factors associated with nonvisualization of a sentinel node (SN) in the axilla area during preoperative lymphoscintigraphy. METHODS: We retrospectively studied 332 women with T0, T1, or T2 <3-cm, N0 invasive breast cancer who underwent a sentinel lymph node biopsy procedure. All patients had intradermal and intraparenchymal injection of 37 MBq (99m)Tc-sulfur colloid in a total volume of 4 x 0.1 mL, above and around the tumor. Anterior and lateral static views were obtained a few minutes and 2-4 h after injection. Surgery was performed the next day. The SNs were localized intraoperatively with the aid of patent blue dye and using a hand-held gamma probe. SNs were analyzed by serial sections stained with hematoxylin-eosin, with the adjacent section stained with anticytokeratin antibodies. Different parameters, such as the number of positive lymph nodes, presence of lymphovascular invasion, tumor size, tumor grade, histology (invasive vs. in situ), prior excisional biopsy, and patient age were analyzed to determine whether they had any significant correlation with nonvisualization of SNs in the axillary area. RESULTS: An axillary SN was successfully visualized on the preoperative lymphoscintigraphy in 302 of 332 patients (90.7%). No axillary drainage was found in 30 patients on the delayed images, even after a second injection of radiocolloid, and 5 of 30 patients showed uptake outside the axillary area. Positive nodes were identified in 86 of 302 patients (28.5%) with successful axillary drainage and in 19 of 30 patients (63.3%) with unsuccessful axillary drainage. More than 4 invaded axillary nodes (P < 0.0001) and the presence of lymphovascular invasion in the breast tumor (P = 0.004) were the only significant variables on univariate analysis, although multivariate analysis showed that only the increased number of invaded nodes was statistically significant. CONCLUSION: Patients with unsuccessful axillary mapping have an increased risk for axillary involvement. PMID- 12902413 TI - Relationship between 18F-FDG uptake and breast density in women with normal breast tissue. AB - Breast density affects the mammographic detectability of breast cancer. The study aimed to evaluate the impact of breast density on the (18)F-FDG uptake of normal breast tissue. METHODS: The study population consisted of 45 women (median age, 54 y; age range, 42-77 y). All underwent whole-body (18)F-FDG PET for various indications other than breast cancer, and all underwent mammography within a mean of 6.6 +/- 4.9 mo of PET. On the basis of mammographic findings, breasts were categorized as extremely dense, heterogeneously dense, primarily fatty, or entirely fatty. Regions of interest were drawn on every PET image in which breast tissue was visualized. Average and peak standardized uptake values (SUVs) were calculated for the left and right breasts. RESULTS: Mammography showed that 20 of the 45 women had heterogeneously dense breasts, 1 had extremely dense breasts, 20 had primarily fatty breasts, and 4 had entirely fatty breasts. In dense breasts, the average SUV was 0.39 +/- 0.05 (right breast) and 0.36 +/- 0.07 (left breast) and the peak SUV was 0.93 +/- 0.16 and 0.89 +/- 0.18, respectively. The average and peak SUVs were significantly lower for primarily fatty breasts than for dense breasts (P < 0.01). Peak and average SUVs of entirely fatty breasts also differed significantly from peak and average SUVs of dense and primarily fatty breasts (P < 0.01). The impact of hormonal status on SUV was significant but less than the impact of breast density. No significant relationship between average SUV or peak SUV and age or serum glucose level was observed. CONCLUSION: Breast density and hormonal status affect the uptake of (18)F-FDG. Dense breasts exhibit, on average, significantly higher (18)F-FDG uptake than do nondense breasts. However, the highest peak SUV observed in dense breasts was 1.39, which is well below the SUV of 2.5 commonly used as a cutoff between benign and malignant tissue. Therefore, breast density is unlikely to affect the ability of (18)F-FDG PET to discriminate between benign and malignant breast lesions. PMID- 12902414 TI - Correction for partial-volume effects on brain perfusion SPECT in healthy men. AB - The limited spatial resolution of SPECT scanners does not allow an exact measurement of the local radiotracer concentration in brain tissue because partial-volume effects (PVEs) underestimate concentration in small structures of the brain. The aim of this study was to determine which brain structures show greater influence of PVEs in SPECT studies on healthy volunteers and to investigate aging effects on SPECT after the PVE correction. METHODS: Brain perfusion SPECT using (99m)Tc-ethylcysteinate dimer was performed in 52 healthy men, 18-86 y old. The regional cerebral blood flow (rCBF) was noninvasively measured using graphical analysis. SPECT images were corrected for PVEs using gray-matter volume, which was segmented from coregistered MR images and convoluted with spatial resolution of SPECT scanners. Absolute rCBF data were measured using a 3-dimensional (3D) stereotactic template for regions of interest on anatomically standardized SPECT. We examined correlation of advancing age with rCBF before and after the PVE correction. To validate the correction method for PVEs, a Hoffman 3D brain phantom experiment was also performed. RESULTS: The PVE correction remarkably reduced the coefficient of variation for SPECT counts in the whole phantom. The PVE correction made the rCBF distribution more homogeneous throughout the brain with less intersubject variation than the original distribution. There were significant negative correlations between age and adjusted rCBF in the bilateral perisylvian and medial frontal areas. These correlations remained significant after the PVE correction. Instead of a positive correlation in the medial temporal structures between age and adjusted rCBF before the PVE correction, the sensorimotor and parietal areas mainly showed positive correlations after the correction. CONCLUSION: SPECT data reflect both brain volume loss and functional changes. Use of the PVE correction in brain perfusion SPECT provides a more accurate determination of rCBF even in healthy volunteers. PMID- 12902415 TI - Severity of bone marrow involvement in patients with Gaucher's disease evaluated by scintigraphy with 99mTc-sestamibi. AB - Gaucher's disease is a lysosomal storage disorder due to a genetically transmitted deficiency of the enzyme glucocerebrosidase. In the most common form of the disease (type 1), accumulation of glucosylceramide in the reticuloendothelial cells of liver, spleen, and bone marrow leads to visceromegaly, anemia, thrombocytopenia, and osteopenia. Skeletal manifestations secondary to infiltration of the bone marrow by Gaucher's cells are detectable by radiography only in advanced stages. Imaging of bone marrow involvement can be performed indirectly by magnetic resonance techniques or by bone marrow scintigraphy with radiocolloids. However, both procedures lack specificity because the normal bone marrow, rather than the pathologic process, is imaged. The aim of this study was to assess the reliability of (99m)Tc-sestamibi scintigraphy for direct evaluation of bone marrow involvement. METHODS: Seventy two patients with type 1 and 2 patients with type 3 Gaucher's disease (35 males, 39 females) were enrolled in the study. The mean age +/- SD was 31.9 +/- 16.5 y (range, 3-76 y), and the average duration of the disease manifestations when performing scintigraphy was 12.95 y (median, 10.5 y; range, 0-44 y). Forty-three of 74 patients had never received enzyme replacement therapy (ERT), whereas 31 patients were already being treated with ERT. (99m)Tc-Sestamibi was injected intravenously (6-8 MBq/kg of body weight) and imaging was recorded at the lower limbs 30 min after injection, at the plateau of tracer accumulation in the involved bone marrow. The scans were evaluated visually, assigning a semiquantitative score based on the extension and intensity of uptake in the bone marrow of the lower limbs (0 = no uptake; 8 = maximum uptake). The scintigraphic score was entered into complex statistical analysis, which included a series of clinical and blood chemistry parameters defining overall severity of the disease. RESULTS: (99m)Tc-Sestamibi scintigraphy showed that 71 of 74 patients had some degree of bone marrow involvement. The scintigraphic score was highly correlated with an overall clinical severity score index (SSI) and with various parameters contributing to the SSI, either positively or negatively. The highest correlation of the scintigraphic score was found with an overall biochemical marker of disease severity (serum chitotriosidase). ERT-naive patients showed high correlation of the scintigraphic score with the clinical SSI, with a radiographically based score, and with serum chitotriosidase. In the ERT-treated patients, the scintigraphic score was correlated with the clinical SSI, with hepatomegaly, and with hemoglobin. CONCLUSION: (99m)Tc-Sestamibi uptake reliably identifies bone marrow infiltration by Gaucher's cells. The scintigraphic score is helpful for defining the severity of bone marrow involvement and for comparing patients. (99m)Tc-Sestamibi scintigraphy, which provides topographic information about the sites involved by the disease, is highly correlated with other parameters of disease severity and appears to correlate with response to ERT. PMID- 12902416 TI - Cholecystokinin cholescintigraphy: methodology and normal values using a lactose free fatty-meal food supplement. AB - The purpose of this investigation was to evaluate the use of a commercially available lactose-free fatty-meal food supplement, as an alternative to sincalide cholescintigraphy, to develop a standard methodology, and to determine normal gallbladder ejection fractions (GBEFs) for this supplement. METHODS: Twenty healthy volunteers all had negative medical histories for hepatobiliary and gallbladder disease, had no personal or family history of hepatobiliary disease, and were not taking any medication known to affect gallbladder emptying. All were prescreened with a complete blood cell count, comprehensive metabolic profile, gallbladder and liver ultrasonography, and conventional cholescintigraphy. Three of the 20 subjects were eliminated from the final analysis because of an abnormality in one of the above studies. RESULTS: After gallbladder filling on conventional cholescintigraphy, the subjects ingested the supplement and an additional 60-min study was acquired. GBEFs were calculated and ranged from 33% to 95% (mean +/- SD, 62.6% +/- 21.3%). Statistical analysis determined the lower range of normal to be 32.6%. Maximal gallbladder emptying occurred between 55 and 60 min. CONCLUSION: A standard methodology and normal GBEFs (> or =33%) were established for supplement-stimulated cholescintigraphy. PMID- 12902417 TI - "USA-Fat": prevalence is related to ambient outdoor temperature-evaluation with 18F-FDG PET/CT. AB - Increased (18)F-FDG activity in fatty tissue has previously been reported with PET/CT. We previously named this activity uptake in supraclavicular area fat ("USA-Fat"). We and others have speculated that this uptake exists in metabolically active brown adipose tissue (BAT). Such tissue might be expected to have varying metabolic activity depending on the ambient temperature. The purpose of this study was to evaluate the frequency of USA-Fat and its relationship to the outdoor temperature. METHODS: Between July 2001 and June 2002, 1,017 consecutive whole-body scans were obtained with a PET/CT scanner and (18)F-FDG for clinical patients. PET images were reviewed for the presence of USA-Fat. RESULTS: USA-Fat was observed in 68 scans obtained from 62 patients (51 female and 11 male). The incidence of USA-Fat was highest, at 13.7%, in January through March, while outside temperatures were low, and was significantly lower, at 4.1%, during the rest of the year. CONCLUSION: The incidence of USA-Fat is clearly increased during the cooler period of the year. This finding suggests that stimulation by cold temperatures increases the frequency with which USA-Fat occurs, supporting underlying BAT as the etiology for this activity. PMID- 12902418 TI - 89Zr immuno-PET: comprehensive procedures for the production of 89Zr-labeled monoclonal antibodies. AB - The use of immuno-PET, the combination of PET with monoclonal antibodies (mAbs), is an attractive option to improve tumor detection and mAb quantification. The long-lived positron emitter (89)Zr has ideal physical characteristics for immuno PET, such as a half-life of 3.27 d, which is compatible with the time needed for intact mAbs to achieve optimal tumor-to-nontumor ratios. Thus far, a major limitation in the use of (89)Zr has been the lack of suitable methods for its stable coupling to mAbs. In this article, practical protocols for reproducible isolation of highly pure (89)Zr and the production of optimal-quality mAb-(89)Zr conjugates are provided. METHODS: (89)Zr was produced by a (p,n) reaction on natural yttrium ((89)Y), isolated with a hydroxamate column, and used for labeling of premodified mAbs. mAbs were premodified with a novel bifunctional derivative of the chelate desferrioxamine B (Df) via a new linker chemistry. To this end, Df was initially succinylated (N-sucDf), temporarily filled with Fe(III), esterified by use of tetrafluorophenol, and then directly coupled to mAbs. Chimeric mAb (cmAb) U36, directed against head and neck cancer, was used for in vitro and in vivo evaluation. The in vitro stability of cmAb U36-N-sucDf (89)Zr was assessed in human serum, and its in vivo behavior was evaluated by biodistribution and PET imaging studies in tumor-bearing nude mice. A cmAb U36-Df (89)Zr conjugate containing a previously described succinimide ring-thioether unit in the linker was used as a reference. RESULTS: (89)Zr was produced in large batches (6.5-13.5 GBq) and isolated with improved radionuclidic purity (>99.99%) and high yield (>94%). The Df-premodified mAbs gave (89)Zr-labeling efficiencies of 80% within 30 min, resulting in conjugates with preserved integrity and immunoreactivity. With respect to stability, the novel cmAb U36-N-sucDf-(89)Zr conjugate appeared to be superior to the reference conjugate. In vivo, the novel conjugate demonstrated selective tumor targeting, and on PET images obtained at 24, 48, and 72 h after injection of this conjugate, small tumors in the range of 19-154 mg were readily visualized. CONCLUSION: Methods were developed for improved purification of the long-lived positron emitter (89)Zr. Moreover, a novel bifunctional Df chelate was synthesized for the reproducible coupling of (89)Zr to mAbs. The suitability of such conjugates to detect millimeter-sized tumors in xenograft-bearing nude mice was demonstrated. PMID- 12902419 TI - Immuno-PET for tumor targeting. PMID- 12902421 TI - Role of lipid-soluble complexes in targeted tumor therapy. AB - Radionuclide therapy remains a promising arsenal against cancer. However, low tumor uptake, high radiation dose to normal organs, and subsequent adverse effects are challenging problems. This study assessed the therapeutic significance of lipid-soluble compounds of (111)In, which passively diffuse through the cell membrane, bind to cytoplasmic components, and remain cell bound until decay. METHODS: Athymic nude mice bearing human colorectal, prostate, or breast cancer received 11.1-14.8 MBq (300-400 micro Ci) (111)In-8 hydroxyquinoline ((111)In-oxine) or (111)In-mercaptopyridine-N-oxide ((111)In Merc) in 200 micro L solution intratumorally through a multihole needle. Tumors in some mice were dissected, and 20- micro m-thick sections were autoradiographed. In additional mice, tumor diameter was measured daily, mice were imaged and weighed, and blood samples were drawn for determination of neutrophil counts for up to 28 d after injection. Some mice were sacrificed at predetermined times for quantitative tissue distribution of (111)In. Additionally, tumor cells were labeled with (111)In-oxine and homogenized, and (111)In associated with cell components was determined using polyacrylamide gel electrophoresis. Radiation dose that could be delivered to adjacent tissues was estimated. The (111)In absorbed dose as a function of radial position r in a 1-g tumor was theoretically compared with those of beta-emitting radionuclides (90)Y and (177)Lu. RESULTS: More than 85% of (111)In remained in tumors, bound to cell cytoplasmic components of apparent molecular weights 250 and 6 kDa. (111)In in tumors was uniformly distributed. Only 2% of the injected (111)In was in the liver, kidneys, and carcass. Statistical analysis showed that on day 28, control tumors grew >100%, whereas treated tumors either had growth arrest or grew only slowly (17%). The estimated radiation dose per megabecquerel (millicurie) injected was 90 Gy/g (9,000 rad/g), of which 64% was from conversion electrons, 16% from Auger electrons, 20% from gamma-photons and x-rays, respectively. Radiation dose to adjacent normal organs was 5%-10% of the radiation dose to the tumor and negligible to the liver and kidneys. Neutrophil counts remained unchanged. Mouse body weight was +/-10% of the initial weight. The radiation dosimetry for (111)In and (177)Lu compared favorably, but not that of (90)Y. CONCLUSION: Treatment is independent of receptor density, heterogeneity, or the hypoxic status of cells. It is applicable to treat all known and accessible tumor types, and it delivers a negligible radiation dose to vital organs and only 5% 10% of the radiation dose to organs adjacent to the tumor. Intratumoral administration of (111)In-oxine appears to be a feasible, effective, safe, and promising treatment for cancer. PMID- 12902420 TI - In vivo evaluation of pretargeted 64Cu for tumor imaging and therapy. AB - Pretargeting involves administration of a tumor-targeting monoclonal antibody (mAb) covalently linked to a molecule having a high-affinity binding site for a rapidly distributed radiolabeled effector molecule. The aim of this study was to compare pretargeting to a conventionally labeled antibody for tumor targeting of the intermediate-lived radionuclide (64)Cu, which has shown promise for PET imaging and radioimmunotherapy of cancer. METHODS: DOTA-biotin (where DOTA is 1,4,7,10-tetraazacyclododecane-N,N',N",N"'-tetraacetic acid) and the intact immunoconjugate DOTA-NR-LU-10 were labeled to high specific activities with (64)Cu, and the serum stabilities and target binding capabilities of each agent were assayed in vitro. Nude mice bearing SW1222 human colorectal carcinoma xenografts were administered (64)Cu-DOTA-biotin, with and without pretreatment with the mAb-streptavidin conjugate NR-LU-10/SA and the synthetic clearing agent Biotin-GalNAc(16), or injected with (64)Cu-DOTA-NR-LU-10. Biodistributions of both agents were obtained from 5 min to 48 h after injection. RESULTS: Both (64)Cu-DOTA-biotin and (64)Cu-DOTA-NR-LU-10 were 100% stable in serum in vitro. (64)Cu-DOTA-biotin exhibited >98% specific binding to immobilized streptavidin, whereas the immunoreactivity of (64)Cu-DOTA-NR-LU-10 averaged nearly 80%. Biodistributions in SW1222-bearing mice showed that NR-LU-10/SA-pretargeted (64)Cu-DOTA-biotin attained a peak tumor uptake of 18.9 percentage injected dose per gram (%ID/g) at 1 h, with concomitant rapid disappearance from blood and renal excretion. In the absence of pretargeting, (64)Cu-DOTA-biotin had very similar biodistribution and clearance properties, except with extremely low nonspecific tumor uptake. In contrast, (64)Cu-DOTA-NR-LU-10 reached 80.3 %ID/g in tumor tissue, after 48 h, whereas blood clearance was considerably slower than pretargeted (64)Cu-DOTA-biotin. Comparison of the time-activity curves for tumor uptake and blood clearance of pretargeted (64)Cu and the (64)Cu-labeled antibody revealed that the maximum tumor accumulations of radioactivity were similar for each agent, 17.9 percentage injected activity per gram (%IA/g) and 20.7 %IA/g, respectively. However, the tumor-to-blood ratio of areas under the curves was 14 times higher for pretargeted (64)Cu-DOTA-biotin because of the substantial increase in blood clearance of the small effector molecule. CONCLUSION: The extremely rapid tumor uptake and blood clearance of pretargeted (64)Cu-DOTA biotin should afford markedly superior PET imaging contrast and therapeutic efficacy, compared with conventionally labeled (64)Cu-DOTA-NR-LU-10. Further comparison of the therapeutic efficacy, toxicity, and dosimetry of these 2 agents is warranted. PMID- 12902422 TI - Meta-analysis of the performance of 18F-FDG PET in primary tumor detection in unknown primary tumors. AB - Detection of the primary tumor has a key role in the management of patients with unknown primary tumors (UPT). The aim of this study was to perform a meta analysis of the literature to evaluate the accuracy of (18)F-FDG PET in primary tumor detection in patients with UPT. METHODS: Systematic methods were used to identify, select, and evaluate the methodologic quality of the studies as well as to summarize the overall findings of sensitivity, specificity, and detection capacity of the primary tumor. The search strategy consisted of identifying studies published between January 1994 and May 2001 indexed in MEDLINE and CANCERLIT. Studies identified by manually searching reference lists of retrieved studies or by reviewing abstracts from recent conference proceedings were also included. Inclusion criteria were studies that evaluated primary tumor detection with (18)F-FDG PET in patients with UPT. Exclusion criteria were duplicated studies or those outdated by subsequent ones. The statistical analysis included 95% confidence intervals (CI) of sensitivity and specificity, both in the pooled data and in the types of studies found. Variation in accuracy between studies was analyzed calculating the natural logarithm of the odds ratio (ln OR) due to study characteristics. Funnel plots of sensitivity and specificity and the summary receiver-operating-characteristic (ROC) curve were also represented. RESULTS: Fifteen studies met the inclusion criteria and were analyzed. Although sample sizes were small, compliance with the methodologic quality criteria was adequate. Heterogeneity analysis showed that differences in the study quality did not correlate with differences in study results. The 95% CI of sensitivity and specificity presented global homogeneity, estimating the sensitivity at 0.87 (95% CI, 0.81-0.92) and the specificity at 0.71 (95% CI, 0.64-0.78). The summary ROC curve showed a good relationship between sensitivity and specificity. The ln OR presented significant values in >75% of the studies. CONCLUSION: (18)F-FDG PET could be useful in patients with UPT for the detection of the primary tumor. (18)F-FDG PET has intermediate specificity and high sensitivity, indicating the existence of few false-negative results, an important feature in the management of oncologic patients that could suggest its utility in the initial stages of the management process. PMID- 12902423 TI - Crucial role for somatostatin receptor subtype 2 in determining the uptake of [111In-DTPA-D-Phe1]octreotide in somatostatin receptor-positive organs. AB - Human somatostatin (SS) receptor (sst)-positive tumors can be visualized by gamma camera scintigraphy after the injection of [(111)In-diethylenetriaminepentaacetic acid (DTPA)-D-Phe(1)] octreotide. Uptake of [(111)In-DTPA-D-Phe(1)]octreotide is dependent on sst-mediated internalization of the radioligand by the tumor cells. Human sst-positive tumors frequently express multiple sst subtypes. In vitro studies have demonstrated that the 5 sst subtypes (sst(1-5)) differentially internalize sst-bound ligand. The present study was performed to evaluate the role of sst(2) in vivo in determining the uptake of [(111)In-DTPA-D Phe(1)]octreotide, as well as of the more "universal" ligand [(111)In-DTPA]SS-14, by sst-positive organs expressing multiple sst subtypes. METHODS: Wild-type and sst(2) knockout mice (n = 4 per treatment group) were injected intravenously with 1 MBq (0.1 micro g) [(111)In-DTPA-D-Phe(1)]octreotide or [(111)In-DTPA]SS-14. After 24 h, the animals were sacrificed and radioactivity in the organs under investigation was determined. In addition, the sst subtype messenger RNA (mRNA) expression pattern in these organs was determined by reverse transcriptase polymerase chain reaction (RT-PCR) analysis. RESULTS: RT-PCR analysis demonstrated the presence of all 5 sst subtype mRNAs in the adrenals and pituitary of wild-type mice but no sst(2) in the knockout mice. The thymus expressed mRNA for sst(2) and sst(4) mRNA in wild-type mice, whereas no sst(2) was detected in knockout mice. In wild-type mice, the in vivo uptake values (in percentage injected dose per gram of tissue) of [(111)In-DTPA-D-Phe(1)]octreotide for the pituitary, adrenals, pancreas, and thymus amounted to 1.2 +/- 0.2, 0.26 +/- 0.03, 0.18 +/- 0.03, and 0.30 +/- 0.05, respectively, in wild-type mice. Compared with wild-type mice, sst(2) knockout mice had dramatically lower uptake values in these organs-lower by 97%, 83%, 96%, and 94%, respectively (P < 0.01 vs. wild type). Comparable differences in the uptake of radioactivity between wild-type and knockout mice were found using [(111)In-DTPA]SS-14 as the radiotracer. Interestingly, in some organs expressing sst(2) mRNA (liver, muscle, and peripheral blood mononuclear cells), no specific binding of [(111)In-DTPA-D Phe(1)]octreotide or [(111)In-DTPA]SS-14 to sst in vivo was found, suggesting that the sst(2) protein expression level was very low in these tissues. CONCLUSION: The uptake of [(111)In-DTPA-D-Phe(1)]octreotide and [(111)In-DTPA]SS 14 in sst-positive organs is determined predominantly by sst(2). PMID- 12902424 TI - Impact of nodal regression on radiation dose for lymphoma patients after radioimmunotherapy. AB - Radioimmunotherapy for non-Hodgkin's lymphoma often results in surprisingly high response rates compared with those expected from estimated absorbed radiation doses. Several factors, including radiobiologic response, selective targeting, and heterogeneous absorbed radiation within the lymphoma, are likely to contribute to the lack of a dose-response relationship. This article investigates the impact of nodal regression on absorbed radiation dose and applies a correction factor to account for its effect. METHODS: The radioactivity in and regression of 37 superficial lymph nodes were measured in 7 non-Hodgkin's lymphoma patients treated with 775-3,450 MBq/m(2) of (131)I-Lym-1 monoclonal antibody. Nodal dimensions were measured with calipers and radioactivity was quantitated using gamma-camera imaging on multiple days after (131)I-Lym-1 injection. Both nodal regression and radioactivity were fit with monoexponential functions. Formulas were developed to account for simultaneous change in nodal mass and radioactivity. All lymph nodes with size and radioactivity measurements, and exponential-fit coefficients of determination of >0.8, were included in the analysis. RESULTS: A 3 orders-of-magnitude node-to-node variation in initial radiopharmaceutical concentration (MBq/g) was observed, with the highest concentrations in the smallest nodes. Reduction in radioactivity as a function of time (biologic half-life) varied by about a factor of 2. In contrast, the rate of nodal regression varied by orders of magnitude, from a 14-h half-time to no regression at all. Five nodes regressed with a half-time that was shorter than their observed effective radiopharmaceutical half-life. Accounting for the effect of nodal regression resulted in dose corrections ranging from 1 (no correction) to a factor of >10, with 70% of nodes requiring a correction factor of at least 20% and >50% of nodes requiring a correction factor of >2. Corrected for nodal regression, 46% of nodes analyzed had absorbed radiation doses of >10 Gy and 32% had doses of >20 Gy. CONCLUSION: These results highlight the importance of accounting for change in mass, particularly tumor regression, when assessing absorbed radiation dose for tissues whose mass changes during the time the radiation dose is being absorbed. The increase in calculated absorbed dose when this change is considered provides better insight into the high nodal response rates observed in non-Hodgkin's lymphoma patients. PMID- 12902425 TI - Biodistribution, radiation dose estimates, and in vivo Pgp modulation studies of 18F-paclitaxel in nonhuman primates. AB - Multidrug resistance (MDR) associated with increased expression and function of the P-glycoprotein (Pgp) efflux pump often causes chemotherapeutic failure in cancer. To provide insight into both the dynamics of the pump and the effects of MDR, we radiolabeled paclitaxel, a substrate for the Pgp pump, with (18)F to study MDR in vivo with PET. We obtained biodistribution and radiation dose estimates for (18)F-paclitaxel (FPAC) in monkeys and studied the effects of a Pgp blocker (XR9576, tariquidar) on FPAC kinetics. METHODS: Paired baseline and Pgp modulation (2 mg/kg XR9576) 4-h whole-body dynamic PET scans were obtained in 3 rhesus monkeys after injection of FPAC. Measured residence times were extrapolated to humans and radiation dose estimates were obtained using MIRDOSE3.1. The postmodulator area under the time-activity curves (AUCs) and Logan plot slopes, a measure of tracer distribution volume (equilibrium tissue-to plasma ratio) that is inversely proportional to tracer efflux, were compared with baseline values to determine changes in FPAC distribution. RESULTS: Cumulative activities of the organs sampled accounted for 80% of the injected dose. The critical organ is gallbladder wall (0.19 mGy/MBq [0.69 rad/mCi]), followed by liver (0.14 mGy/MBq [0.52 rad/mCi]); the effective dose is 0.022 mSv/MBq (0.083 rem/mCi). XR9576 preinfusion changed the Logan plot slope for liver by +104% (P = 0.02), lung by +87% (P = 0.11), and kidney by -14% (P = 0.08). Changes in the mean AUC (normalized to the plasma AUC) were +54% (P = 0.08), +97% (P = 0.04), and -12% (P = 0.02), respectively, for liver, lung, and kidney. No significant difference was found in the metabolite-corrected plasma AUC (normalized to the injected dose) between the baseline and XR9576 modulator studies (P = 0.69). CONCLUSION: Under Radioactive Drug Research Committee guidelines, 266 MBq (7.2 mCi) FPAC can be administered to humans up to 3 times a year. The increase in FPAC accumulation in liver and lung after XR9576 is consistent with Pgp inhibition and demonstrates the potential of FPAC to evaluate MDR. PMID- 12902426 TI - Performance of a brain PET camera based on anger-logic gadolinium oxyorthosilicate detectors. AB - A high-sensitivity, high-resolution brain PET scanner ("G-PET") has been developed. This scanner is similar in geometry to a previous brain scanner developed at the University of Pennsylvania, the HEAD Penn-PET, but the detector technology and electronics have been improved to achieve enhanced performance. METHODS: This scanner has a detector ring diameter of 42.0 cm with a patient aperture of 30.0 cm and an axial field of view of 25.6 cm. It comprises a continuous light-guide that couples 18,560 (320 x 58 array) 4 x 4 x 10 mm(3) gadolinium oxyorthosilicate (GSO) crystals to 288 (36 x 8 array) 39-mm photomultiplier tubes in a hexagonal arrangement. The scanner operates only in 3 dimensional (3D) mode because there are no interplane septa. Performance measurements on the G-PET scanner were made following National Electrical Manufacturers Association NU 2-2001 procedures for most measurements, although NU 2-1994 procedures were used when these were considered more appropriate for a brain scanner (e.g., scatter fraction and counting-rate performance measurements). RESULTS: The transverse and axial resolutions near the center are 4.0 and 5.0 mm, respectively. At a radial offset of 10 cm, these numbers deteriorate by approximately 0.5 mm. The absolute sensitivity of this scanner measured with a 70-cm long line source is 4.79 counts per second (cps)/kBq. The scatter fraction measured with a line source in a 20-cm-diameter x 19-cm-long cylinder is 39% (for a lower energy threshold of 410 keV). For the same cylinder, the peak noise equivalent counting rate is 60 kcps at an activity concentration of 7.4 kBq/mL (0.20 micro Ci/mL), whereas the peak true coincidence rate is 132 kcps at an activity concentration of 14 kBq/mL (0.38 micro Ci/mL). Images from the Hoffman brain phantom as well as (18)F-FDG patient scans illustrate the high quality of images acquired on the G-PET scanner. CONCLUSION: The G-PET scanner attains the goal of high performance for brain imaging through the use of an Anger-logic GSO detector design with continuous optical coupling. This detector design leads to good energy resolution, which is needed in 3D imaging to minimize scatter and random coincidences. PMID- 12902427 TI - Comparison of 2-dimensional and 3-dimensional 82Rb myocardial perfusion PET imaging. AB - We compared 2-dimensional (2D) and 3-dimensional (3D) (82)Rb PET imaging in 3 different experiments: in a realistic heart-thorax phantom, in a uniformity resolution phantom, and in 14 healthy volunteers. METHODS: A nonuniform heart thorax phantom was filled with 111 MBq of (82)Rb injected into the left ventricular (LV) wall. In the LV wall of the cardiac phantom, 3 inserts-1, 2, and 3 cm in diameter-were placed to simulate infarcts. A standard rest cardiac PET imaging protocol in 2D and 3D modes was used. Following the same protocol, a uniformity-resolution phantom with uniformly distributed activity of 1,998 MBq and 740 MBq of (82)Rb in water was used to obtain 2D PET images and 3D PET images, respectively. All 2D volunteer studies were performed by injecting 2,220 MBq of (82)Rb intravenously. For half the volunteers, 3D studies were performed with a high dose (HD) (2,220 MBq) of (82)Rb; for the remainder of the 3D studies, a low dose (LD) (740 MBq) of (82)Rb was used. In the 2D and LD 3D studies, there was a delay of 2 min and 3 min, respectively, followed by a 6-min acquisition. In the HD 3D volunteer studies, there was a delay of 5 min followed by a 6-min acquisition. Circumferential profiles of the short-axis slices and the contrast of the inserts were used to evaluate the cardiac phantom PET images. The transaxial slices from the uniformity-resolution phantom were evaluated by visual inspection and by measuring uniformity. The human studies were evaluated by measuring the contrast between LV wall and LV cavity, using linear profiles and visual analysis. RESULTS: In the cardiac phantom study, circumferential profiles for the 2D and 3D images were similar. The contrast values for the 1-, 2-, and 3 cm inserts in the 2D study were 0.19 +/- 0.03, 0.34 +/- 0.05, and 0.61 +/- 0.03, respectively. The respective contrast values in the 3D study were 0.15 +/- 0.02, 0.36 +/- 0.04, and 0.52 +/- 0.05. In the uniformity-resolution phantom study, the coefficients of variation, calculated for a representative uniform slice, were 5.3% and 7.6% for the 2D and 3D studies, respectively. For the 7 volunteers on whom HD 3D was used, the mean 2D contrast was 0.33 +/- 0.08 and the mean HD 3D contrast was 0.35 +/- 0.08 (P = not statistically significant). For the other 7 volunteers, on whom LD 3D was used, the mean 2D contrast was 0.39 +/- 0.06 and the mean LD 3D contrast was 0.39 +/- 0.10 (P = not statistically significant). In the tomographic slices, the 2D and 3D images and polar plots were similar. CONCLUSION: When obtained with a PET system having a high counting-rate performance, 2D and 3D (82)Rb PET cardiac images are comparable. LD 3D imaging can make (82)Rb PET cardiac imaging more affordable. PMID- 12902428 TI - A Bayesian 3-compartment model for 99mTc-MAG3 clearance. AB - Because recent reports have questioned the traditional 2-compartment model for calculating tracer clearance after a single intravenous injection, a 3 compartment model was evaluated in this study. METHODS: Bayesian statistics were used, which facilitated curve fitting by treating all subjects simultaneously. (99m)Tc-Mercaptoacetyltriglycine clearance data from 154 adults and 109 children were measured at several centers, typically 6-9 plasma samples spanning 5-90 min, and fitted by 2- and 3-compartment Bayesian models. RESULTS: Clearance estimates were found to be systematically lower for the 3-compartment model than for the 2 compartment model. A single-sample procedure based on the 3-compartment model was found to eliminate most of the known discrepancy between formulas based on single injection and continuous-infusion reference methods. CONCLUSION: A 3-compartment model led to lower and probably more accurate clearance estimates than the conventional 2-compartment model. A new single-sample method is presented, based on the 3-compartment model as reference standard. PMID- 12902429 TI - The birth of nuclear medicine instrumentation: Blumgart and Yens, 1925. AB - In 1925, Hermann Blumgart performed the first diagnostic procedure using radioactive indicators on humans; this first is well recognized. Less well recognized is the fact that Blumgart and his coworker Otto C. Yens, then a medical student, developed the first instrumentation used in a diagnostic procedure involving radioactive indicators. The instrumentation, a modified Wilson cloud chamber, turned out to be the detector most suitable for their purpose. Blumgart also showed remarkable foresight in outlining the requirements both for a satisfactory indicator (tracer) and for a satisfactory detector- requirements that still hold true today. The Blumgart-Yens modified cloud chamber was the birth of nuclear medicine instrumentation. PMID- 12902430 TI - Specificity of 99mTc-ciprofloxacin imaging. PMID- 12902431 TI - An advance in small-cell lung cancer treatment--more or less. PMID- 12902432 TI - From bedside to bench: treatment for rare blood disorder yields clues to disease mechanism. PMID- 12902434 TI - Terminology for reporting adverse events updated. PMID- 12902435 TI - Stat bite: Incidence of lung and bronchus cancer by ethnicity, 1996-2000. PMID- 12902436 TI - The Will Rogers phenomenon: Roping the effects of a new cancer staging system. PMID- 12902437 TI - Breast cancer group targets 'shareholder activism' at Avon. PMID- 12902438 TI - Big science can be big success, IOM report concludes. PMID- 12902440 TI - National Institutes of Health State-of-the-Science Conference Statement: Symptom Management in Cancer: Pain, Depression, and Fatigue, July 15-17, 2002. AB - BACKGROUND: Despite advances in early detection and effective treatment, cancer remains one of the most feared diseases. Among the most common side effects of cancer and treatments for cancer are pain, depression, and fatigue. Although research is producing increasingly hopeful insights into the causes and cures for cancer, efforts to manage the side effects of the disease and its treatments have not kept pace. The challenge that faces us is how to increase awareness of the importance of recognizing and actively addressing cancer-related distress. The National Institutes of Health (NIH) convened a State-of-the-Science Conference on Symptom Management in Cancer: Pain, Depression, and Fatigue to examine the current state of knowledge regarding the management of pain, depression, and fatigue in individuals with cancer and to identify directions for future research. Specifically, the conference examined how to identify individuals who are at risk for cancer-related pain, depression, and/or fatigue; what treatments work best to address these symptoms when they occur; and what is the best way to deliver interventions across the continuum of care. State-of-the-Science Process: A non-advocate, non-Federal, 14-member panel of experts representing the fields of oncology, radiology, psychology, nursing, public health, social work, and epidemiology prepared the statement. In addition, 24 experts in medical oncology, geriatrics, pharmacology, psychology, and neurology presented data to the panel and to the conference audience during the first 1.5 days of the conference. The panel then prepared its statement, addressing the five predetermined questions and drawing on submitted literature, the speakers' presentations, and discussions held at the conference. The statement was presented to the conference audience, followed by a press conference to allow the panel to respond to questions from the media. After its release at the conference, the draft statement was made available on the Internet. The panel's final statement is available at http://consensus.nih.gov. CONCLUSIONS: The panel concluded that the available evidence supports a variety of interventions for treating cancer patients' pain, depression, and fatigue. Clinicians should routinely use brief assessment tools to ask patients about pain, depression, and fatigue and to initiate evidence based treatments. Assessment should include discussion about common symptoms experienced by cancer patients, and these discussions should continue over the duration of the illness. Impediments to effective symptom management in cancer patients can arise from different sources and interactions among providers, patients and their families, and the health care system. Numerous factors could interfere with adequate symptom management. Among these factors are incomplete effectiveness of some treatments, a lack of sufficient knowledge regarding effective treatment strategies, patient reluctance to report symptoms to caregivers, a belief that such symptoms are simply a part of the cancer experience that must be tolerated, and inadequate coverage and reimbursement for some treatments. Additional research is needed on the definition, occurrence, the treatment of pain, depression, and fatigue, alone and in combination, in adequately funded prospective studies. The panel also concluded that the state of the science in cancer symptom management should be reassessed periodically. PMID- 12902441 TI - Randomized phase III trial of paclitaxel, etoposide, and carboplatin versus carboplatin, etoposide, and vincristine in patients with small-cell lung cancer. AB - BACKGROUND: Paclitaxel administered in combination with a topoisomerase-II inhibitor (such as etoposide) and carboplatin is an effective and safe first-line treatment for patients with small-cell lung cancer (SCLC). We conducted a randomized phase III multicenter trial to determine whether paclitaxel plus etoposide plus carboplatin improves the outcome of patients with primary SCLC relative to standard chemotherapy (carboplatin, etoposide, and vincristine). METHODS: Between January 1998 and December 1999, 614 patients with SCLC stages I IV were randomly assigned to the standard arm (309 patients) or the experimental arm (305 patients). Treatment courses were repeated every 21 days for a maximum of six courses. All patients were evaluated for response rate, survival, and toxicities every two courses. The primary endpoint was survival. Survival curves were estimated with the Kaplan-Meier method and compared using the log-rank test. All statistical tests were two-sided. RESULTS: A total of 608 patients were evaluable for all endpoints (standard arm 307 patients, experimental arm 301 patients). The hazard ratio [HR] of death for patients receiving the standard treatment was statistically significantly higher than that for patients receiving the experimental treatment (HR = 1.22, 95% confidence interval [CI] = 1.03 to 1.45; P =.024). Progression-free survival was also statistically significantly shorter for patients in the standard arm relative to that of patients in the experimental arm (HR = 1.21, 95% CI = 1.03 to 1.42). There were no differences in the response rates (complete and partial combined) to the treatments (standard arm: 69.4%, 95% CI = 63.9% to 74.5%; experimental arm: 72.1%, 95% CI = 66.7% to 77.1%; difference = 2.7%, 95% CI = 4.5% to 9.9%). Rates of severe grade of anemia, leukocytopenia, neutropenia, and thrombocytopenia were lower in the experimental arm than in the standard arm. CONCLUSION: Patients with previously untreated SCLC who received paclitaxel, etoposide, and carboplatin showed improved overall and progression-free survival and less frequent hematologic toxicities than those who received the standard therapy. PMID- 12902442 TI - Specific antibody levels at the cervix during the menstrual cycle of women vaccinated with human papillomavirus 16 virus-like particles. AB - BACKGROUND: In early-phase trials, a human papillomavirus 16 (HPV16) virus-like particle (VLP) vaccine has been shown to be well tolerated, immunogenic, and protective against HPV16 in women, most of whom were taking oral contraceptives. Previous studies have not determined whether HPV immunization results in specific antibody levels in the human genital tract or whether these levels might vary during contraceptive or ovulatory cycles. Therefore, we determined the levels of total and specific antibodies in the cervical secretions of women who had been immunized with HPV16 VLPs and examined the influence of the menstrual cycle and oral contraceptive use on these levels. METHODS: Two groups of women were immunized, seven who were taking oral contraceptives and 11 who were ovulating. After seroconversion, serum and cervical secretions were collected twice weekly for 5 weeks. Total immunoglobulins (IgG and IgA) and vaccine-specific IgGs were determined by enzyme-linked immunosorbent assay. Nonparametric statistical analyses were used to determine the statistical significance of differences in IgG levels between groups, and correlations between serum- and cervical-specific IgG levels were determined by the Spearman correlation coefficient. RESULTS: All participants developed detectable titers of anti-HPV16 VLP IgGs in their cervical secretions after immunization. The cervical titers of specific IgG and total IgGs and IgAs among participants in the contraceptive group were relatively constant throughout the contraceptive cycle. In contrast, the cervical titers of specific IgG and total IgGs and IgAs among participants in the ovulatory group varied during the menstrual cycle, being highest during the proliferative phase, decreasing approximately ninefold around ovulation, and increasing approximately threefold during the luteal phase. Serum- and cervical-specific IgG levels were correlated (r =.86) in women in the contraceptive group but not in women in the ovulatory group (r =.27). CONCLUSIONS: The relatively high titer of anti-HPV16 antibodies at the cervix is promising in terms of vaccine efficacy; however, the decrease in antibody titer around ovulation raises the possibility that the HPV16 VLP vaccine might be less effective during the peri-ovulatory phase. PMID- 12902443 TI - Tumor-selective induction of apoptosis and the small-molecule immune response modifier imiquimod. AB - BACKGROUND: The incidence of nonmelanoma skin cancer, basal cell carcinomas (BCCs), and squamous cell carcinomas (SCCs) is increasing, representing a major medical and economic problem. Imiquimod, a topical small-molecule immune response modifier, has shown efficacy toward BCC and actinic keratoses in clinical trials. Imiquimod acts both indirectly, via cytokine-mediated stimulation of cellular immune responses, and directly, through unknown mechanisms against tumor cells. We examined the mechanism by which imiquimod induces apoptosis in cancer cells. METHODS: Apoptosis was assessed by enzyme-linked immunosorbent assay, western blot analysis, and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assays in five SCC cell lines, HaCaT cells (a spontaneously immortalized human keratinocyte cell line), and normal keratinocytes treated with imiquimod, with its analog resiquimod, or with neither. Expression of death receptors, caspases, and cytochrome c in the apoptotic signaling cascade was analyzed using western blot and flow cytometric analyses. The functional relevance of imiquimod-induced cytochrome c release was assessed by transfection of HaCaT cells with Bcl-2. Apoptosis in BCCs in vivo was assessed by TUNEL assays of imiquimod-treated and untreated tumors from three patients. Differences between treated and untreated cells and tumors were determined using a two-tailed Student's t test. RESULTS: Imiquimod, but not resiquimod, induced apoptosis in all SCC cell lines and HaCaT cells. This induction involved activation of several caspases and Bcl-2-dependent cytosolic translocation of cytochrome c but was independent of the membrane-bound death receptors Fas, tumor necrosis factor related apoptosis-inducing ligand (TRAIL)-R1-R4 receptors, and tumor necrosis factor-R1 and -R2 receptors. Topical application of imiquimod to BCC tumors in vivo induced apoptosis. CONCLUSION: Imiquimod has the potential to induce apoptosis in skin cancer cells, possibly by circumventing mechanisms developed by malignant tumors to resist apoptotic signals. PMID- 12902444 TI - Integrin beta3 Leu33Pro homozygosity and risk of cancer. AB - BACKGROUND: Increased tumor cell expression of integrins containing the beta3 subunit is associated with increased progression to invasive tumors, whereas inhibition of beta3 integrin expression and/or function may reduce tumor growth and metastasis. The Leu33Pro polymorphism of the beta3 subunit modulates the function of alpha(IIb)beta3 integrin. We examined whether this polymorphism influences cancer risk. METHODS: Using participants (n = 9242) from the Copenhagen City Heart Study with 24 years of follow-up and endpoints from the Danish Cancer Registry, we assessed the risk of all cancers and of 27 cancer types in individuals who carry the Leu33Pro polymorphism (heterozygotes and homozygotes) relative to those without the polymorphism (non-carriers). Relative risks (RRs) of cancer and 95% confidence intervals (CIs) were calculated by Cox proportional hazards regression analysis. Differences in cumulative cancer incidence (per 10 000 person-years) were tested using log-rank statistics. Statistical tests were two-sided. RESULTS: Among the participants, 70.0% were non carriers, 27.3% were heterozygotes, and 2.7% were homozygotes. We detected 1296 participants with a first cancer. Cumulative incidences in non-carriers, heterozygotes, and homozygotes were 81, 83, and 112, respectively (homozygotes versus non-carriers, P =.02). The age-adjusted RR of all cancers in homozygotes relative to non-carriers was 1.4 (95% CI = 1.1 to 1.9). Incidences in non carriers, heterozygotes, and homozygotes were 3, 4, and 16 for ovarian cancer; 19, 24, and 36 for breast cancer; and 2, 3, and 7 for melanoma (homozygotes versus non-carriers; P =.002, P =.06, and P =.03, respectively). The age-adjusted RR in homozygotes relative to non-carriers was 4.7 (95% CI = 1.6 to 14) for ovarian cancer, 1.9 (95% CI = 1.0 to 3.7) for breast cancer, and 3.5 (95% CI = 1.1 to 12) for melanoma. Adjustment for other cancer risk factors did not alter these results. Heterozygotes did not differ from non-carriers with respect to cancer risk. CONCLUSION: Individuals homozygous for the Leu33Pro polymorphism of the beta3 integrin subunit have an increased cancer risk. PMID- 12902445 TI - Phytoestrogen intake and endometrial cancer risk. AB - BACKGROUND: The development of endometrial cancer is largely related to prolonged exposure to unopposed estrogens. Phytoestrogens (i.e., weak estrogens found in plant foods) may have antiestrogenic effects. We evaluated the associations between dietary intake of seven specific compounds representing three classes of phytoestrogens (isoflavones, coumestans, and lignans) and the risk of endometrial cancer. METHODS: In a case-control study from the greater San Francisco Bay Area, we collected dietary information from 500 African American, Latina, and white women aged 35-79 years who were diagnosed with endometrial cancer between 1996 and 1999 and from 470 age- and ethnicity-matched control women identified through random-digit dialing. Unconditional logistic regression analyses were used to estimate odds ratios (ORs) and 95% confidence intervals (CIs). RESULTS: Isoflavone (OR = 0.59, 95% CI = 0.37 to 0.93 for the highest versus lowest quartile of exposure) and lignan (OR = 0.68, 95% CI = 0.44 to 1.1) consumptions were inversely related to the risk of endometrial cancer. These associations were slightly stronger in postmenopausal women (OR = 0.44, 95% CI = 0.26 to 0.77 and OR = 0.57, 95% CI = 0.34 to 0.97 for isoflavones and lignans, respectively). Obese postmenopausal women consuming relatively low amounts of phytoestrogens had the highest risk of endometrial cancer (OR = 6.9, 95% CI = 3.3 to 14.5 compared with non-obese postmenopausal women consuming relatively high amounts of isoflavones); however, the interaction between obesity and phytoestrogen intake was not statistically significant. CONCLUSION: Some phytoestrogenic compounds, at the levels consumed in the typical American-style diet, are associated with reduced risk of endometrial cancer. PMID- 12902446 TI - T-cell homeostasis in breast cancer survivors with persistent fatigue. AB - Approximately 30% of women successfully treated for breast cancer suffer persistent fatigue of unknown origin. Recent studies linking inflammatory processes to central nervous system-mediated fatigue led us to examine cellular immune system status in 20 fatigued breast cancer survivors and 19 matched non fatigued breast cancer survivors. Fatigued survivors, compared with non-fatigued survivors, had statistically significantly increased numbers of circulating T lymphocytes (mean 31% increase, 95% confidence interval [CI] = 6% to 56%; P =.015 by two-sided analysis of variance [ANOVA]), with pronounced elevation in the numbers of CD4+ T lymphocytes (mean 41% increase, 95% CI = 15% to 68%; P =.003 by two-sided ANOVA) and CD56+ effector T lymphocytes (mean 52% increase, 95% CI = 4% to 99%; P =.027 by two-sided ANOVA). These changes were independent of patient demographic and treatment characteristics. Absolute numbers of B cells, natural killer cells, granulocytes, and monocytes were not altered. The increased numbers of circulating T cells correlated with elevations in the level of serum interleukin 1 receptor antagonist (for CD3+ cells, r =.56 and P =.001; for CD3+/CD4+ cells, r =.68 and P<.001, by Spearman rank correlation). Results of this study suggest that persistent fatigue in breast cancer survivors might be associated with a chronic inflammatory process involving the T-cell compartment. These results require confirmation in a larger study that is specifically designed to address this hypothesis. PMID- 12902448 TI - Re: Adjuvant chemotherapy in patients with early-stage ovarian cancer. PMID- 12902450 TI - Re: Leptin--a growth factor in normal and malignant breast cells and for normal mammary gland development. PMID- 12902452 TI - Re: A randomized, placebo-controlled trial of zoledronic acid in patients with hormone-refractory metastatic prostate carcinoma. PMID- 12902453 TI - Re: Short-interval follow-up mammography: are we doing the right thing? PMID- 12902456 TI - Was induction of HIV-1 through TLR9? PMID- 12902457 TI - FOXO transcription factors as regulators of immune homeostasis: molecules to die for? AB - Regulation of phosphatidylinositol 3-kinase (PI3K) activity has been demonstrated to be critical for correct lymphocyte function. The molecular targets of this lipid kinase have been the subject of extensive research, and many functional effects of PI3K activation are thought to be mediated by the serine-threonine kinase protein kinase B (PKB/c-akt). Genetic analyses in the nematode worm Caenorhabditis elegans have identified a novel PI3K-regulated signaling pathway that regulates organism lifespan through inhibition of a Forkhead (FOX) transcription factor, DAF-16. Recent studies have subsequently revealed an evolutionarily conserved signaling module in higher eukaryotes in which PKB can directly phosphorylate and inactive a family of Forkhead box class O (FOXO) transcription factors. Phosphorylation results in nuclear exclusion and inhibition of transcription. FOXO transcription factors have been found to play critical roles in regulation of proliferation, apoptosis and control of oxidative stress. This occurs through both activation and repression of target gene expression by multiple mechanisms. Here the regulation and function of these transcription factors is discussed with specific relevance to immune homeostasis. A greater understanding of the regulation and function of this signaling pathway in lymphocytes may provide novel therapeutic opportunities for immune diseases. PMID- 12902458 TI - Toll-like receptors in health and disease: complex questions remain. PMID- 12902459 TI - Cutting edge: invariant V alpha 14 NKT cells are required for allergen-induced airway inflammation and hyperreactivity in an experimental asthma model. AB - Airway hyperreactivity (AHR), eosinophilic inflammation with a Th2-type cytokine profile, and specific Th2-mediated IgE production characterize allergic asthma. In this paper, we show that OVA-immunized Jalpha18(-/-) mice, which are exclusively deficient in the invariant Valpha14(+) (iValpha14), CD1d-restricted NKT cells, exhibit impaired AHR and airway eosinophilia, decreased IL-4 and IL-5 production in bronchoalveolar lavage fluid, and reduced OVA-specific IgE compared with wild-type (WT) littermates. Adoptive transfer of WT iValpha14 NKT cells fully reconstitutes the capacity of Jalpha18(-/-) mice to develop allergic asthma. Also, specific tetramer staining shows that OVA-immunized WT mice have activated (CD69(+)) iValpha14 NKT cells. Importantly, anti-CD1d mAb treatment blocked the ability of iValpha14 T cells to amplify eosinophil recruitment to airways, and both Th2 cytokine and IgE production following OVA challenge. In conclusion, these findings clearly demonstrate that iValpha14 NKT cells are required to participate in allergen-induced Th2 airway inflammation through a CD1d-dependent mechanism. PMID- 12902460 TI - Cutting edge: the B cell chemokine CXC chemokine ligand 13/B lymphocyte chemoattractant is expressed in the high endothelial venules of lymph nodes and Peyer's patches and affects B cell trafficking across high endothelial venules. AB - While CCR7 ligands direct T cell trafficking into lymph nodes (LNs) and Peyer's patches (PPs), chemokines that regulate B cell trafficking across high endothelial venules (HEVs) remain to be fully elucidated. Here we report that CXC chemokine ligand (CXCL)13 (B lymphocyte chemoattractant) is detected immunohistologically in the majority of HEVs in LNs and PPs of nonimmunized mice. Systemically administered anti-CXCL13 Ab bound to the surface of approximately 50% of HEVs in LNs and PPs, but not to other types of blood vessels, indicating that CXCL13 is expressed in the HEV lumen. In CXCL13-null mice, B cells rarely adhered to PP HEVs, whereas T cells did efficiently. Superfusion of CXCL13-null PPs with CXCL13 restored the luminal presentation of CXCL13 and also B cell arrest in PP HEVs at least partially. Collectively, these results indicate that CXCL13 expressed in the HEV lumen plays a crucial role in B cell trafficking into secondary lymphoid tissues such as PPs. PMID- 12902461 TI - Cutting edge: SR-PSOX/CXC chemokine ligand 16 mediates bacterial phagocytosis by APCs through its chemokine domain. AB - SR-PSOX and CXC chemokine ligand (CXCL)16, which were originally identified as a scavenger receptor and a transmembrane-type chemokine, respectively, are indicated to be identical. In this study, we demonstrate that membrane-bound SR PSOX/CXCL16 mediates adhesion and phagocytosis of both Gram-negative and Gram positive bacteria. Importantly, our prepared anti-SR-PSOX mAb, which suppressed chemotactic activity of SR-PSOX, significantly inhibited bacterial phagocytosis by human APCs including dendritic cells. Various scavenger receptor ligands inhibited the bacterial phagocytosis of SR-PSOX. In addition, the recognition specificity for bacteria was determined by only the chemokine domain of SR PSOX/CXCL16. Thus, SR-PSOX/CXCL16 may play an important role in facilitating uptake of various pathogens and chemotaxis of T and NKT cells by APCs through its chemokine domain. PMID- 12902462 TI - Cutting edge: induced indoleamine 2,3 dioxygenase expression in dendritic cell subsets suppresses T cell clonal expansion. AB - In mice, immunoregulatory APCs express the dendritic cell (DC) marker CD11c, and one or more distinctive markers (CD8alpha, B220, DX5). In this study, we show that expression of the tryptophan-degrading enzyme indoleamine 2,3 dioxygenase (IDO) is selectively induced in specific splenic DC subsets when mice were exposed to the synthetic immunomodulatory reagent CTLA4-Ig. CTLA4-Ig did not induce IDO expression in macrophages or lymphoid cells. Induction of IDO completely blocked clonal expansion of T cells from TCR transgenic mice following adoptive transfer, whereas CTLA4-Ig treatment did not block T cell clonal expansion in IDO-deficient recipients. Thus, IDO expression is an inducible feature of specific subsets of DCs, and provides a potential mechanistic explanation for their T cell regulatory properties. PMID- 12902463 TI - Germinal centers regulate human Th2 development. AB - In the present study we demonstrate that all CD4(+) T cells in human tonsil expressing the Th2-selective receptor chemoattractant receptor-homologous molecule expressed on Th2 cells (CRTH2) also 1) express high levels of CXCR5, and 2) display a transitional CD45RA/RO phenotype and consistently do not produce significant amounts of cytokines when immediately analyzed ex vivo. Hence, they represent precursors of Th2 effector cells, a conclusion confirmed by their robust production of IL-4, IL-5, and IL-13, but not IFN-gamma, after in vitro activation. CD4(+) T cells, which express only intermediate levels of CXCR5, instead develop into IFN-gamma-producing cells under identical culture conditions, thus establishing a correlation between relative levels of CXCR5 expression and the acquired cytokine profile. Because CXCR5 is critically involved in follicular localization, the results suggest that these CRTH2(+) Th2 cells preferentially develop their cytokine-producing phenotype within germinal centers (GCs), whereas extrafollicular differentiation instead promotes Th1 development. In support for this proposal, we show that T cells with an intermediate expression of CXCR5 can be forced to also produce IL-4 and IL-13 if cultured with allogenic GC B cells. Finally, we demonstrate that the previously described CD57(+) GC T cells also express high levels of CXCR5 but instead of comprising a Th2 precursor, they represent anergized T cells. Taken together, these data suggest that GCs and B cells regulate CD4(+) T cell differentiation in a finely tuned fashion, either by promoting differentiation of Th2 cells, which apparently leave the lymphoid tissue before evolving a cytokine-producing phenotype, or by furnishing T cell unresponsiveness. PMID- 12902464 TI - Lymphocyte accumulation in the spleen of retinoic acid receptor-related orphan receptor gamma-deficient mice. AB - The hormone nuclear receptor retinoic acid receptor-related orphan receptor gamma (RORgamma) plays important roles in thymocyte development and lymphoid organogenesis. RORgamma and its thymus-specific isoform RORgammat are expressed in the thymus, but not in the spleen and bone marrow (BM). However, RORgamma(-/-) mice have 2- to 3-fold more splenocytes than wild-type controls due to an accumulation of conventional resting B lymphocytes. The increase in B lymphocytes in RORgamma(-/-) mice is caused neither by abnormal B cell development in the BM nor by an obvious defect in the peripheral T cell compartment. Furthermore, analyses of BM chimeras using either RORgamma(-/-) or recombinase-activating gene 2(-/-) mice as recipients and wild-type or RORgamma(-/-) mice as donors, respectively, demonstrate that the splenic microenvironment of RORgamma(-/-) mice is defective, since wild-type T and B lymphocytes accumulated in these chimeric mice. In addition, T lymphocyte homeostasis was altered due to a lowered thymic output in RORgamma(-/-) mice. Collectively, these results suggest that RORgamma regulates lymphocyte homeostasis at multiple levels. PMID- 12902466 TI - Plasma cell survival is mediated by synergistic effects of cytokines and adhesion dependent signals. AB - Recent results suggest that plasma cell longevity is not an intrinsic capacity, but depends on yet unknown factors produced in their environment. In this study, we show that the cytokines IL-5, IL-6, TNF-alpha, and stromal cell-derived factor 1alpha as well as signaling via CD44 support the survival of isolated bone marrow plasma cells. The cytokines IL-7 and stem cell factor, crucially important for early B cell development, do not mediate plasma cell survival, indicating that plasma cells and early B cells have different survival requirements. As shown in IL-6-deficient mice, IL-6 is required for a normal induction, but not for the maintenance of plasma cell responses in vivo, indicating that the effects of individual survival factors are redundant. Optimal survival of isolated plasma cells requires stimulation by a combination of factors acting synergistically. These results strongly support the concept that plasma cell survival depends on niches in which a combination of specific signals, including IL-5, IL-6, stromal cell-derived factor-1alpha, TNF-alpha, and ligands for CD44, provides an environment required to mediate plasma cell longevity. PMID- 12902467 TI - Differential localization and function of ADP-ribosylation factor-6 in anergic human T cells: a potential marker for their identification. AB - Anergy is a state of immunologic tolerance in which T cells are viable but incapable of responding to antigenic stimulation. Recent data indicate that anergic cells have a distinct gene expression program that determines their unique function. In this study we show that anergic human T cells selectively express the small GTPase ADP-ribosylation factor-6 (ARF6), which is involved in membrane traffic and regulation of the cortical actin cytoskeleton. ARF6 was expressed in the GTP-bound form that localizes at the plasma membrane, resulting in a distinct morphologic appearance of anergic cells. Forced expression of ARF6 GTP in Jurkat T cells prevented TCR-mediated reorganization of cortical actin, extracellular signal-regulated kinase1/2 activation, and IL-2 transcription. Forced expression of ARF6-GTP in primary human T cells inhibited extracellular signal-regulated kinase1/2 activation and proliferative responses. Importantly, T cells with the distribution pattern of ARF6-GTP were detected in peripheral blood, suggesting that anergic T cells may constitutively exist in vivo. PMID- 12902465 TI - Mechanisms of ganglioside inhibition of APC function. AB - Gangliosides shed by tumor cells exert potent inhibitory effects on cellular immune responses. Here we have studied ganglioside inhibition of APC function. When human monocytes were preincubated in 50 micro M highly purified ganglioside G(D1a), pulsed with tetanus toxoid (TT), and washed, the expected Ag-induced proliferative response of autologous normal T cells added to these monocytes was inhibited by 81%. Strikingly, there was also almost complete (92%) and selective inhibition of the up-regulation of the monocyte costimulatory molecule CD80, while I-CAM-1, LFA-3, HLA-DR, and CD86 expression were unaffected. Purified LPS stimulated monocytes that had been preincubated in G(D1a) likewise showed inhibition of CD80 up-regulation (59%) as well as down-regulation of CD40 (54%) and impaired release of IL-12 and TNF-alpha (reduced by 59 and 51%). G(D1a) preincubated human dendritic cells (DC) were also affected. They had reduced constitutive expression of CD40 (33%) and CD80 (61%), but not CD86, and marked inhibition of release of IL-6 (72%), IL-12 (70%), and TNF-alpha (46%). Even when pulsed with TT, these ganglioside-preincubated DC remained deficient in costimulatory molecule expression and cytokine secretion and were unable to induce a normal T cell proliferative response to TT. Finally, significant inhibition of nuclear localization of NF-kappaB proteins in activated DC suggests that disruption of NF-kappaB activation may be one mechanism contributing to ganglioside interference with APC expression of costimulatory molecules and cytokine secretion, which, in turn, may diminish antitumor immune responses. PMID- 12902468 TI - Functional dissection of lupus susceptibility loci on the New Zealand black mouse chromosome 1: evidence for independent genetic loci affecting T and B cell activation. AB - In previous work, we demonstrated linkage between a broad region on New Zealand Black (NZB) chromosome 1 and increased costimulatory molecule expression on B cells and autoantibody production. In this study, we produced C57BL/6 congenic mice with homozygous NZB chromosome 1 intervals of differing lengths. We show that both B6.NZBc1(35-106) (numbers denote chromosomal interval length) and B6.NZBc1(85-106) mice produce IgG anti-nuclear autoantibodies, but B6.NZBc1(35 106) mice develop significantly higher titers of autoantibodies and more severe renal disease than B6.NZBc1(85-106) mice. Cellular analysis of B6.NZBc1(85-106) mice revealed splenomegaly and increased numbers of memory T cells. In addition to these features, B6.NZBc1(35-106) mice had altered B and T cell activation with increased expression of CD69, and for B cells, costimulatory molecules and MHC. Introduction of an anti-hen egg white lysozyme Ig transgene, as a representative nonself-reactive Ig receptor, onto the B6.NZBc1(35-106) background corrected the B cell activation phenotype and led to dramatic normalization of splenomegaly and T cell activation, but had little impact on the increased proportion of memory T cells. These findings indicate that there are multiple lupus susceptibility genes on NZB chromosome 1, and that although B cell defects play an important role in lupus pathogenesis in these mice, they act in concert with T cell activation defects. PMID- 12902469 TI - Selective inhibition of T cell activation via CD147 through novel modulation of lipid rafts. AB - The plasma membrane is compartmentalized into microdomains and the association/dissociation of receptors and signaling molecules with/from these membrane domains is a major principle for regulation of signal transduction. By following the reorganization of microdomains on living cells and performing biochemical studies, we show that Ab targeting of the T cell activation associated Ag CD147 prevents TCR stimulation-dependent reorganization and clustering of microdomains. Triggering CD147 induces a displacement of the GPI anchored coreceptors CD48 and CD59 from microdomains in human T lymphocytes. This perturbation of microdomains is accompanied by a selective inhibition of TCR mediated T cell proliferation. The CD147-inhibited cells secret normal levels of IL-2 but acquire reduced amounts of the IL-2 receptor alpha-chain CD25. These results indicate that negative regulating signals can modulate microdomains and suggest a general mechanism for inhibition of receptor signaling. PMID- 12902470 TI - Blastocyst MHC, a putative murine homologue of HLA-G, protects TAP-deficient tumor cells from natural killer cell-mediated rejection in vivo. AB - Blastocyst MHC is a recently identified mouse MHC class Ib gene, which is selectively expressed in blastocyst and placenta, and may be the mouse homolog of HLA-G gene the products of which have been implicated in protection of fetal trophoblasts from maternal NK cells and evasion of some tumor cells from NK cell attack. In this study, we identified two blastocyst MHC gene transcripts encoding a full-length alpha-chain (bc1) and an alternatively spliced form lacking the alpha2 domain (bc2), which may be homologous to HLA-G1 and HLA-G2, respectively. Both placenta and a teratocarcinoma cell line predominantly expressed the bc2 transcript. When these cDNAs were expressed in TAP-deficient RMA-S or TAP sufficient RMA cells, only bc1 protein was expressed on the surface of RMA cells, but both bc1 and bc2 proteins were retained in the cytoplasm of RMA-S cells. Significantly, the RMA-S cells expressing either bc1 or bc2 were protected from lysis by NK cells in vitro. This protection was at least partly mediated by up regulation of Qa-1(b) expression on the surface of RMA-S cells, which engaged the CD94/NKG2A inhibitory receptor on NK cells. More importantly, the bc1- or bc2 expressing RMA-S cells were significantly protected from NK cell-mediated rejection in vivo. These results suggested a role for blastocyst MHC in protecting TAP-deficient trophoblasts and tumor cells from NK cell attack in vivo. PMID- 12902471 TI - CCR3 expression induced by IL-2 and IL-4 functioning as a death receptor for B cells. AB - We report that CCR3 is not expressed on freshly isolated peripheral and germinal B cells, but is up-regulated after stimulation with IL-2 and IL-4 (approximately 98% CCR3(+)). Ligation of CCR3 by eotaxin/chemokine ligand (CCL) 11 induces apoptosis in IL-2- and IL-4-stimulated primary CD19(+) (approximately 40% apoptotic cells) B cell cultures as well as B cell lines, but has no effect on chemotaxis or cell adhesion. Freshly isolated B cells express low levels of CD95 and CD95 ligand (CD95L) (19 and 21%, respectively). Expression is up-regulated on culture in the presence of a combination of IL-2, IL-4, and eotaxin/CCL11 (88% CD95 and 84% CD95L). We therefore propose that ligation of such newly induced CCR3 on peripheral and germinal B cells by eotaxin/CCL11 leads to the enhanced levels of CD95 and CD95L expression. Ligation of CD95 by its CD95L expressed on neigboring B cells triggers relevant death signaling pathways, which include an increase in levels of Bcl-2 expression, its functional activity, and the release of cytochrome c from the mitochondria into the cytosol. These events initiate a cascade of enzymatic processes of the caspase family, culminating in programmed cell death. Interaction between CCR3 and eotaxin/CCL11 may, besides promoting allergic reactions, drive activated B cells to apoptosis, thereby reducing levels of Ig production, including IgE, and consequently limit the development of the humoral immune response. The apoptotic action of eotaxin/CCL11 suggests a therapeutic modality in the treatment of B cell lymphoma. PMID- 12902472 TI - Synoviocyte-mediated expansion of inflammatory T cells in rheumatoid synovitis is dependent on CD47-thrombospondin 1 interaction. AB - Fibroblast-like synoviocytes (FLS) from patients with rheumatoid arthritis elicit spontaneous proliferation of autologous T cells in an HLA-DR and CD47 costimulation-dependent manner. T cell costimulation through CD47 is attributed to specific interaction with thrombospondin-1 (TSP1), a CD47 ligand displayed on FLS. CD47 binding by FLS has broad biological impact that includes adhesion and the triggering of specific costimulatory signals. TSP1(+) FLS are highly adhesive to T cells and support their aggregation and growth in situ. Long-term cultures of T cells and FLS form heterotypic foci that are amenable to propagation without exogenous growth factors. T cell adhesion and aggregate formation on TSP1(+) FLS substrates are inhibited by CD47-binding peptides. In contrast, FLS from arthroscopy controls lack adhesive or T cell growth-promoting activities. CD47 stimulation transduces a costimulatory signal different from that of CD28, producing a gene expression profile that included induction of ferritin L chain, a component of the inflammatory response. Ferritin L chain augments CD3-induced proliferation of T cells. Collectively, these results demonstrate the active role of FLS in the recruitment, activation, and expansion of T cells in a CD47 dependent manner. Because TSP1 is abundantly expressed in the rheumatoid synovium, CD47-TSP1 interaction is proposed to be a key component of an FLS/T cell regulatory circuit that perpetuates the inflammatory process in the rheumatoid joint. PMID- 12902473 TI - Identifying MHC class I epitopes by predicting the TAP transport efficiency of epitope precursors. AB - We are able to make reliable predictions of the efficiency with which peptides of arbitrary lengths will be transported by TAP. The pressure exerted by TAP on Ag presentation thus can be assessed by checking to what extent MHC class I (MHC-I) presented epitopes can be discriminated from random peptides on the basis of predicted TAP transport efficiencies alone. Best discriminations were obtained when N-terminally prolonged epitope precursor peptides were included and the contribution of the N-terminal residues to the score were down-weighted in comparison with the contribution of the C terminus. We provide evidence that two factors may account for this N-terminal down-weighting: 1) the uncertainty as to which precursors are used in vivo and 2) the coevolution in the C-terminal sequence specificities of TAP and other agents in the pathway, which may vary among the various MHC-I alleles. Combining predictions of MHC-I binding affinities with predictions of TAP transport efficiency led to an improved identification of epitopes, which was not the case when predictions of MHC-I binding affinities were combined with predictions of C-terminal cleavages made by the proteasome. PMID- 12902474 TI - RNA interference shows critical requirement for NF-kappa B p50 in the production of IL-12 by human dendritic cells. AB - Specific NF-kappaB/Rel proteins regulate murine dendritic cell (DC) survival, differentiation, and activation, but little is known of their role in human cells because of limited loss-of-function analyses. RNA interference (RNAi) is a mechanism to effectively silence gene expression via sequence-specific double stranded small interfering RNAs (siRNAs). RNAi was used to assess the role of the p50 (NF-kappaB1) protein in the maturation and activation of cultured human monocyte-derived DC (MoDC). Transfection of cultured MoDC with siRNAs reduced p50 mRNA and protein levels in a specific, dose-dependent, and time-dependent manner. Basal or maturation-induced expression of HLA-DR and costimulatory molecules were not affected, whereas transcription of the IL-12 p40 gene and the secretion of IL 12alphabeta were reduced. Such MoDC induced less IFN-gamma production by T cells in MLR. This is the first report of RNAi-induced phenotype in human primary DC with a method that caused no measurable toxicity or type-I IFN response. siRNAs appear useful for the study of signaling pathways in immune cells, revealing a pivotal requirement for p50 in MoDC for IL-12 production and induction of optimal type-1 immune responses. PMID- 12902475 TI - Incomplete activation of CD4 T cells by antigen-presenting transitional immature B cells: implications for peripheral B and T cell responsiveness. AB - B cells leave the bone marrow as transitional B cells. Transitional B cells represent a target of negative selection and peripheral tolerance, both of which are abrogated in vitro by mediators of T cell help. In vitro, transitional and mature B cells differ in their responses to B cell receptor ligation. Whereas mature B cells up-regulate the T cell costimulatory molecule CD86 (B7.2) and are activated, transitional B cells do not and undergo apoptosis. The ability of transitional B cells to process and present Ag to CD4 T cells and to elicit protective signals in the absence of CD86 up-regulation was investigated. We report that transitional B cells can process and present Ag as peptide:MHC class II complexes. However, their ability to activate T cells and elicit help signals from CD4-expressing Th cells was compromised compared with mature B cells, unless exogenous T cell costimulation was provided. A stringent requirement for CD28 costimulation was not evident in interactions between transitional B cells and preactivated CD4-expressing T cells, indicating that T cells involved in vivo in an ongoing immune response might rescue Ag-specific transitional B cells from negative selection. These data suggest that during an immune response, immature B cells may be able to sustain the responses of preactivated CD4(+) T cells, while being unable to initiate activation of naive T cells. Furthermore, the ability of preactivated, but not naive T cells to provide survival signals to B cell receptor-engaged transitional immature B cells argues that these B cells may be directed toward activation rather than negative selection when encountering Ag in the context of a pre-existing immune response. PMID- 12902476 TI - KIR2DL4 (CD158d) genotype influences expression and function in NK cells. AB - The expression and function of the NK cell receptor KIR2DL4 are controversial. Two common alleles of the transmembrane domain of KIR2DL4 exist. The 10A allele with 10 adenines at the end of the transmembrane exon encodes a full length receptor, whereas the 9A allele has only 9 adenines resulting in a frame shift which in turn generates a stop codon early in the first cytoplasmic exon. The possibility that the 10A and 9A alleles might result in differences in expression and function of KIR2DL4 was explored using mAbs to KIR2DL4. Transfection experiments with cDNA from the 10A and 9A alleles revealed significant membrane expression only with the protein encoded by the 10A allele. Analysis of peripheral blood NK cells demonstrated that only in subjects with at least one 10A allele was cell surface expression of KIR2DL4 detectable, and then only on the minor CD56(bright) NK cell subset. The major CD56(dim) NK cell subset did not cell surface express KIR2DL4 but, interestingly, did so after in vitro culture. Functional analysis using cultured NK cells in redirected lysis assays demonstrated that KIR2DL4 is an activating receptor for NK cells with at least one 10A allele. No significant activity was detected for NK cells generated from subjects homozygous for the 9A allele. These data show that genotype influences cell surface expression and function of KIR2DL4 which may account for reported differences in KIR2DL4 expression and function. PMID- 12902477 TI - NKT cells from normal and tumor-bearing human livers are phenotypically and functionally distinct from murine NKT cells. AB - A major group of murine NK T (NKT) cells express an invariant Valpha14Jalpha18 TCR alpha-chain specific for glycolipid Ags presented by CD1d. Murine Valpha14Jalpha18(+) account for 30-50% of hepatic T cells and have potent antitumor activities. We have enumerated and characterized their human counterparts, Valpha24Vbeta11(+) NKT cells, freshly isolated from histologically normal and tumor-bearing livers. In contrast to mice, human NKT cells are found in small numbers in healthy liver (0.5% of CD3(+) cells) and blood (0.02%). In contrast to those in blood, most hepatic Valpha24(+) NKT cells express the Vbeta11 chain. They include CD4(+), CD8(+), and CD4(-)CD8(-) cells, and many express the NK cell markers CD56, CD161, and/or CD69. Importantly, human hepatic Valpha24(+) T cells are potent producers of IFN-gamma and TNF-alpha, but not IL-2 or IL-4, when stimulated pharmacologically or with the NKT cell ligand, alpha galactosylceramide. Valpha24(+)Vbeta11(+) cell numbers are reduced in tumor bearing compared with healthy liver (0.1 vs 0.5%; p < 0.04). However, hepatic cells from cancer patients and healthy donors release similar amounts of IFN gamma in response to alpha-galactosylceramide. These data indicate that hepatic NKT cell repertoires are phenotypically and functionally distinct in humans and mice. Depletions of hepatic NKT cell subpopulations may underlie the susceptibility to metastatic liver disease. PMID- 12902478 TI - Induced dendritic cell differentiation of chronic myeloid leukemia blasts is associated with down-regulation of BCR-ABL. AB - Although differentiation of leukemic blasts to dendritic cells (DC) has promise in vaccine strategies, the mechanisms underlying this differentiation and the differences between leukemia and normal progenitor-derived DC are largely undescribed. In the case of chronic myeloid leukemia (CML), understanding the relationship between the induction of DC differentiation and the expression of the BCR-ABL oncogene has direct relevance to CML biology as well as the development of new therapeutic approaches. We now report that direct activation of protein kinase C (PKC) by the phorbol ester PMA in the BCR-ABL(+) CML cell line K562 and primary CML blasts induced nonterminal differentiation into cells with typical DC morphology (cytoplasmic dendrites), characteristic surface markers (MHC class I, MHC class II, CD86, CD40), chemokine and transcription factor expression, and ability to stimulate T cell proliferation (equivalent to normal monocyte-derived DC). PKC-induced differentiation was associated with down regulation of BCR-ABL mRNA expression, protein levels, and kinase activity. This down-regulation appeared to be signaled through the mitogen-activated protein kinase pathway. Therefore, PKC-driven differentiation of CML blasts into DC-like cells suggests a potentially novel strategy to down-regulate BCR-ABL activity, yet raises the possibility that CML-derived DC vaccines will be less effective in presenting leukemia-specific Ags. PMID- 12902479 TI - Either of the CD45RB and CD45RO isoforms are effective in restoring T cell, but not B cell, development and function in CD45-null mice. AB - The protein tyrosine phosphatase CD45 is expressed as a series of isoforms whose tissue and differentiation stage specificity is broadly conserved in evolution. CD45 has been shown to be an important regulator of a variety of functions in many different hemopoietic lineages. We have chosen an in vivo genetic complementation strategy to investigate the differential functions between isoforms. In this study, we report the characterization of transgenic mice which express the isoforms CD45RO or CD45RB as their only CD45 molecules, at a variety of expression levels and in the majority of hemopoietic lineages. Both CD45RO and CD45RB isoforms reconstitute thymocyte development in a CD45-null mouse background when expressed above a threshold level. The resulting mature T cells populate the peripheral lymphoid organs where they are found at normal frequency. Both CD45RO and CD45RB isoforms also permit T cell function in the periphery, although the threshold for normal function here appears to be set higher than in the thymus. In contrast, neither isoform is capable of fully restoring peripheral B cell maturation, even at levels approaching those in heterozygous CD45(+/-) mice in which maturation is normal. In vitro activation of B cells by Ag-receptor stimulation is only minimally complemented by these CD45RO and CD45RB transgenes. Our results suggest that CD45 isoforms play unique roles which differ between the T and B lineages. PMID- 12902481 TI - IL-18 stimulates the proliferation and IFN-gamma release of CD4+ T cells in the chicken: conservation of a Th1-like system in a nonmammalian species. AB - The phylogeny of Th1 and Th2 subsets has not been characterized mainly due to the limited information regarding cytokines in nonmammalian vertebrates. In this study, we characterize a Th1-like regulatory system focusing on the IL-18 regulated IFN-gamma secretion. Stimulation of splenocytes with chicken IL-18 induced high levels of IFN-gamma secretion. Depletion of either macrophages or CD4(+) T cells from the splenocyte cultures caused unresponsiveness to IL-18. In contrast, PBL were unresponsive to IL-18 in the presence or absence of macrophages, but IFN-gamma secretion was stimulated by suboptimal anti-TCR cross linking combined with IL-18. Splenocytes from five different chicken lines responded equally well to the IL-18 treatment. LSL chicken splenocytes, however, responded only to IL-18 when stimulated either with optimal TCR cross-linking alone or suboptimal TCR cross-linking combined with IL-18. IL-18 not only induced IFN-gamma secretion, but also stimulated splenocyte proliferation. This IL-18 induced proliferation was compared with the effects observed with IL-2. Both cytokines activated the splenocytes as demonstrated by increased size and MHC class II Ag up-regulation in the case of IL-18. Phenotypic analyses following 6 days of culture revealed that IL-2 mainly affected the proliferation of CD8(+) cells, whereas IL-18 had an opposite effect and stimulated the proliferation of CD4(+) cells. Taken together, these results demonstrate the conservation of Th1 like proinflammatory responses in the chicken; they characterize IL-18 as a major growth factor of CD4(+) T cells and identify two distinct mechanisms of IL-18 induced IFN-gamma secretion. PMID- 12902480 TI - Break of neonatal Th1 tolerance and exacerbation of experimental allergic encephalomyelitis by interference with B7 costimulation. AB - Ig-PLP1 is an Ig chimera expressing proteolipid protein-1 (PLP1) peptide corresponding to aa residues 139-151 of PLP. Newborn mice given Ig-PLP1 in saline on the day of birth and challenged 7 wk later with PLP1 peptide in CFA develop an organ-specific neonatal immunity that confers resistance against experimental allergic encephalomyelitis. The T cell responses in these animals comprise Th2 cells in the lymph node and anergic Th1 lymphocytes in the spleen. Intriguingly, the anergic splenic T cells, although nonproliferative and unable to produce IFN gamma or IL-4, secrete significant amounts of IL-2. In this work, studies were performed to determine whether costimulation through B7 molecules plays any role in the unusual form of splenic Th1 anergy. The results show that engagement of either B7.1 or B7.2 with anti-B7 Abs during induction of EAE in adult mice that were neonatally tolerized with Ig-PLP1 restores and exacerbates disease severity. At the cellular level, the anergic splenic T cells regain the ability to proliferate and produce IFN-gamma when stimulated with Ag in the presence of either anti-B7.1 or anti-B7.2 Ab. However, such restoration was abolished when both B7.1 and B7.2 molecules were engaged simultaneously, indicating that costimulation is necessary for reactivation. Surprisingly, both anti-B7.1 and anti-B7.2 Abs triggered splenic dendritic cells to produce IL-12, a key cytokine required for restoration of the anergic T cells. Thus, recovery from neonatally induced T cell anergy requires B7 molecules to serve double functions, namely, costimulation and induction of cytokine production by APCs. PMID- 12902482 TI - T cell-intrinsic requirement for NF-kappa B induction in postdifferentiation IFN gamma production and clonal expansion in a Th1 response. AB - NF-kappaB/Rel transcription factors are linked to innate immune responses and APC activation. Whether and how the induction of NF-kappaB signaling in normal CD4(+) T cells regulates effector function are not well-understood. The liberation of NF kappaB dimers from inhibitors of kappaB (IkappaBs) constitutes a central checkpoint for physiologic regulation of most forms of NF-kappaB. To investigate the role of NF-kappaB induction in effector T cell responses, we targeted inhibition of the NF-kappaB/Rel pathway specifically to T cells. The Th1 response in vivo is dramatically weakened when T cells defective in their NF-kappaB induction (referred to as IkappaBalpha(DeltaN) transgenic cells) are activated by a normal APC population. Analyses in vivo, and IL-12-supplemented T cell cultures in vitro, reveal that the mechanism underlying this T cell-intrinsic requirement for NF-kappaB involves activation of the IFN-gamma gene in addition to clonal expansion efficiency. The role of NF-kappaB in IFN-gamma gene expression includes a modest decrease in Stat4 activation, T box expressed in T cell levels, and differentiation efficiency along with a more prominent postdifferentiation step. Further, induced expression of Bcl-3, a trans-activating IkappaB-like protein, is decreased in T cells as a consequence of NF-kappaB inhibition. Together, these findings indicate that NF-kappaB induction in T cells regulates efficient clonal expansion, Th1 differentiation, and IFN-gamma production by Th1 lymphocytes at a control point downstream from differentiation. PMID- 12902483 TI - The human fascin gene promoter is highly active in mature dendritic cells due to a stage-specific enhancer. AB - Dendritic cells (DC), regarded as the most efficient APCs of the immune system, are capable of activating naive T cells. Thus, DC are primary targets in immunotherapy. However, little is known about gene regulation in DC, and for efficient transcriptional targeting of human DC, a suitable promoter is still missing. Recently, we successfully used the promoter of the murine actin-bundling protein fascin to transcriptionally target DC by DNA vaccination in mice. In this study, we report on isolation of the human fascin promoter and characterization of its regulatory elements. The actively expressed gene was distinguished from a conserved inactive genomic locus and a continuous region of 14 kb covering the gene and 3 kb of 5'-flanking sequences was subcloned, sequenced, and analyzed for regulatory elements. Regulatory sequences were found solely in the 5'-flanking promoter region. The promoter exerted robust activity in DC and a fascin-positive neuronal cell line, but not in the fascin-negative cells tested. Notably, promoter activity in DC markedly increased with maturation of DC. By progressive 5' deletion, we identified a core promoter region, harboring a putative GC box, a composite cAMP responsive element/AP-1 binding site and a TATA box. By internal deletion, we demonstrated functional importance of either regulatory element. Furthermore, we identified a more distal stage-specific enhancer region also containing silencer elements. Taken together, the human fascin promoter allows for transcriptional targeting of mature DC and represents a promising tool for immunotherapy. To our knowledge, this study reports for the first time on promoter activity in human monocyte-derived DC. PMID- 12902484 TI - Inhibitory coreceptors activated by antigens but not by anti-Ig heavy chain antibodies install requirement of costimulation through CD40 for survival and proliferation of B cells. AB - Ag-induced B cell proliferation in vivo requires a costimulatory signal through CD40, whereas B cell Ag receptor (BCR) ligation by anti-Ig H chain Abs, such as anti-Ig micro H chain Ab and anti-Ig delta H chain Ab, alone induces proliferation of B cells in vitro, even in the absence of CD40 ligation. In this study, we demonstrate that CD40 signaling is required for survival and proliferation of B cells stimulated by protein Ags in vitro as well as in vivo. This indicates that the in vitro system represents B cell activation in vivo, and that protein Ags generate BCR signaling distinct from that by anti-Ig H chain Abs. Indeed, BCR ligation by Ags, but not by anti-Ig H chain Abs, efficiently phosphorylates the inhibitory coreceptors CD22 and CD72. When these coreceptors are activated, anti-Ig H chain Ab-stimulated B cells can survive and proliferate only in the presence of CD40 signaling. Conversely, treatment of Ag-stimulated B cells with anti-CD72 mAb blocks CD72 phosphorylation and induces proliferation, even in the absence of CD40 signaling. These results strongly suggest that activation of B cells by anti-Ig H chain Abs involves their ability to silence the inhibitory coreceptors, and that the inhibitory coreceptors install requirement of CD40 signaling for survival and proliferation of Ag-stimulated B cells. PMID- 12902485 TI - Alpha 3 domain mutants of peptide/MHC class I multimers allow the selective isolation of high avidity tumor-reactive CD8 T cells. AB - The goal of adoptive T cell therapy in cancer is to provide effective antitumor immunity by transfer of selected populations of tumor Ag-specific T cells. Transfer of T cells with high TCR avidity is critical for in vivo efficacy. In this study, we demonstrate that fluorescent peptide/MHC class I multimeric complexes incorporating mutations in the alpha3 domain (D227K/T228A) that abrogate binding to the CD8 coreceptor can be used to selectively isolate tumor Ag-specific T cells of high functional avidity from both in vitro expanded and ex vivo T cell populations. Sorting, cloning, and expansion of alpha3 domain mutant multimer-positive CD8 T cells enabled rapid selection of high avidity tumor reactive T cell clones. Our results are relevant for ex vivo identification and isolation of T cells with potent antitumor activity for adoptive T cell therapy. PMID- 12902486 TI - Reduced dosage of Bruton's tyrosine kinase uncouples B cell hyperresponsiveness from autoimmunity in lyn-/- mice. AB - The development of autoimmunity is correlated with heightened sensitivity of B cells to B cell Ag receptor (BCR) cross-linking. BCR signals are down-regulated by Lyn, which phosphorylates inhibitory receptors. lyn(-/-) mice have reduced BCR signaling thresholds and develop autoantibodies, glomerulonephritis, splenomegaly due to myeloid hyperplasia, and increased B-1 cell numbers. Bruton's tyrosine kinase (Btk), a critical component of BCR signaling pathways, is required for autoantibody production in lyn(-/-) mice. It is unclear whether Btk mediates autoimmunity at the level of BCR signal transduction or B cell development, given that lyn(-/-)Btk(-/-) mice have a severe reduction in conventional B and B-1 cell numbers. To address this issue, we crossed a transgene expressing a low dosage of Btk (Btk(low)) in B cells to lyn(-/-)Btk(-/-) mice. Conventional B cell populations were restored to levels similar to those in lyn(-/-) mice. These cells were as hypersensitive to BCR cross-linking as lyn(-/-) B cells as measured by proliferation, Ca(2+) flux, and activation of extracellular signal-regulated kinase and Akt. However, lyn(-/-)Btk(low) mice did not produce anti-ssDNA, anti dsDNA, anti-histone, or anti-histone/DNA IgM or IgG. They also lacked B-1 cells and did not exhibit splenomegaly. Thus, B cell hyperresponsiveness is insufficient for autoimmunity in lyn(-/-) mice. These studies implicate B-1 and/or myeloid cells as key contributors to the lyn(-/-) autoimmune phenotype. PMID- 12902487 TI - Apoptosis by neglect of CD4+ Th cells in granulomas: a novel effector mechanism involved in the control of egg-induced immunopathology in murine schistosomiasis. AB - In infection with Schistosoma mansoni, parasite eggs precipitate an intrahepatic granulomatous and fibrosing inflammation that is mediated by CD4(+) Th cells. Compared with CBA mice, C57BL/6 mice develop smaller granulomas composed of cells that exhibit reduced proliferative responses to schistosome egg Ags. In the present study, we investigated CD4(+) T cell apoptosis as a possible mechanism that could account for this subdued response. We found throughout the course of several infection weeks a markedly higher proportion of apoptotic CD4(+) T cells in granulomas from C57BL/6 mice than in those from CBA mice ex vivo; the apoptosis further increased upon cell cultivation in vitro. Activation-induced cell death or CD8(+) T cells failed to account for the enhanced apoptosis as infected Fas-, Fas ligand,- and CD8-deficient mice exhibited similar apoptosis to that seen in wild-type counterparts. However, a strikingly lower IL-2 production by schistosome egg Ag-stimulated C57BL/6 granuloma and mesenteric lymph node cells suggested the possibility of apoptosis due to growth factor deprivation. Indeed, the CD4(+) T cell apoptosis was significantly reversed by addition of rIL 2 in vitro, or by injection of rIL-2 in vivo, which also resulted in significant exacerbation of granulomatous inflammation. These findings indicate that apoptosis by neglect can represent a significant means of controlling CD4(+) T cells that mediate the immunopathology in schistosomiasis. PMID- 12902488 TI - Pathogen-associated molecular patterns sensitize macrophages to Fas ligand induced apoptosis and IL-1 beta release. AB - Antigenic stimulation activates T cells and simultaneously destines them to die by Fas-mediated apoptosis. In this study, we demonstrated that various pathogen associated molecular patterns up-regulated Fas expression in macrophages and sensitized them specifically to Fas ligand (FasL), but not to other apoptosis inducing agents such as TNF-alpha, etoposide (VP-16), and staurosporine. Toll like receptor, NF-kappaB, and p38 mitogen-activated protein kinase mediated these responses. LPS stimulation induced the expression of Fas, caspase 8, cellular FLIP Bfl-1/A1, and Bcl-x, but not FasL, TNFR p55, Bak, Bax, and Bad at the transcriptional level. Thus, LPS selectively induced the expression of apoptotic molecules of the Fas death pathway (except for cellular FLIP) and antiapoptotic molecules of the mitochondrial death pathway. However, the kinetics of macrophage disappearance following Escherichia coli-induced peritonitis was similar between wild-type and Fas-deficient mice, suggesting that Fas is not essential for the turnover of activated macrophages in T cell-independent inflammation. In contrast, LPS-activated macrophages produced a large amount of IL-1beta upon FasL stimulation. Thus, unlike the activation-induced cell death of T cells, the sensitization of macrophages to FasL by pathogen-associated molecular patterns seems to be a proinflammatory rather than an anti-inflammatory event. PMID- 12902489 TI - A catalytically inactive form of protein kinase C-associated kinase/receptor interacting protein 4, a protein kinase C beta-associated kinase that mediates NF kappa B activation, interferes with early B cell development. AB - Protein kinase C-associated kinase (PKK)/receptor interacting protein 4 (RIP4) is a protein kinase C (PKC) beta-associated kinase that links PKC to NF-kappaB activation. The kinase domain of PKK is similar to that of RIP, RIP2, and RIP3. We show in this study that PKK is expressed early during lymphocyte development and can be detected in common lymphoid progenitor cells. Targeting of a catalytically inactive version of PKK to lymphoid cells resulted in a marked impairment in pro-B cell generation in the bone marrow. Although peripheral B cell numbers were markedly reduced, differentiation into follicular and marginal zone B cells was not defective in these mice. B-1a and B-1b B cells could not be detected in these mice, but this might be a reflection of the overall defect in B cell production observed in these animals. In keeping with a possible link to PKCbeta, peripheral B cells in these mice exhibit a defect in anti-IgM-mediated proliferation. These studies suggest that PKK may be required early in B cell development and for BCR-mediated B cell proliferation. PMID- 12902490 TI - Studies of structure-activity relations of complement inhibitor compstatin. AB - Compstatin, a 13-mer cyclic peptide, is a novel and promising inhibitor of the activation of the complement system. In our search for a more active analog and better understanding of structure-functions relations, we designed a phage displayed random peptide library based on previous knowledge of structure activity relations, in which seven amino acids deemed necessary for structure and activity were kept fixed while the remaining six were optimized. Screening of this library against C3 identified four binding clones. Synthetic peptides corresponding to these clones revealed one analog, called acetylated Ile(1)Leu/His(9)Trp/Thr(13)Gly triple replacement analog of compstatin corresponding to clone 640 (Ac-I1L/H9W/T13G), which was more active than compstatin. This newly identified peptide had 4-fold higher activity when compared with the originally isolated form of compstatin and 1.6-fold higher activity when compared with acetylated compstatin (Ac-compstatin). The structures of Ac-I1L/H9W/T13G and Ac-compstatin were studied by nuclear magnetic resonance, compared with the structure of compstatin, and found to be very similar. The binding of Ac-I1L/H9W/T13G and the equally active acetylated analog with His(9)Ala replacement (Ac-H9A) to C3 was evaluated by surface plasmon resonance, which suggested similarity in their binding mechanism but difference when compared with Ac-compstatin. Compensatory effects of flexibility outside the beta turn and tryptophan ring stacking may be responsible for the measured activity increase in Ac-I1L/H9W/T13G and acetylated analog with His(9)Ala replacement and the variability in binding mechanism compared with Ac-compstatin. These data demonstrate that tryptophan is a key amino acid for activity. Finally, the significance of the N-terminal acetylation was examined and it was found that the hydrophobic cluster at the linked termini of compstatin is essential for binding to C3 and for activity. PMID- 12902491 TI - New programming of IL-4 receptor signal transduction in activated T cells: Stat6 induction and Th2 differentiation mediated by IL-4Ralpha lacking cytoplasmic tyrosines. AB - Signaling by the IL-4 receptor alpha-chain (IL-4Ralpha) is a key determinant of the development of the Th2 lineage of effector T cells. Studies performed in tissue culture cell lines have indicated that tyrosines of the IL-4Ralpha cytoplasmic tail are necessary for the induction of Stat6, a transcription factor required for Th2 differentiation. Surprisingly, we have found that in activated T cells, IL-4Ralpha chains lacking all cytoplasmic tyrosines promote induction of this IL-4-specific transcription factor and efficient commitment to the Th2 lineage. Mutagenesis of a tyrosine-free cytoplasmic tail identifies a requirement for the serine-rich ID-1 region in this new program of IL-4R signal transduction observed in activated T cells. Additional findings suggest that an extracellular signal-regulated kinase pathway can be necessary and sufficient for the ability of such tyrosine-free IL-4Ralpha chains to mediate Stat6 induction. These results provide novel evidence that the molecular mechanisms by which a cytokine specifically induces a Stat transcription factor can depend on the activation state of T lymphoid cells. Furthermore, the data suggest that one pathway by which such new programming may be achieved is mediated by extracellular signal regulated mitogen-activated protein kinases. PMID- 12902493 TI - Potential role of NKG2D/MHC class I-related chain A interaction in intrathymic maturation of single-positive CD8 T cells. AB - The nonclassical MHC class I molecule MHC class I-related chain A (MICA) interacts with the NKG2D receptor expressed at the surface of most peripheral CD8 T cells, gammadelta T cells, and NK cells. We investigated the role of MICA-NKG2D interactions in the selection or maturation of the T cell repertoire within the thymus using MICA tetramers and anti-MICA mAbs. MICA tetramers identified a small population of late stage CD8 single-positive, CD45RA(+) CD62L(+) CCR7(+) CD69(-) thymocytes, a phenotype compatible with that of fully mature CD8(+) cells ready to emigrate to the periphery as naive cells. MICA molecules were expressed in the outer layer of Hassal's corpuscles within the medulla of normal thymus. In thymomas, an overexpression of MICA in cortical and medullar epithelial cells was observed. This was associated with a decreased percentage of NKG2D-positive thymocytes, which expressed a less mature phenotype than in normal thymus. These results indicate that CD8(+) thymocytes up-regulate NKG2D as they complete their developmental program before leaving the thymic medulla to seed the periphery, and identify NKG2D as a potential regulator of the developmental processes in T cells that are essential for immune homeostasis. PMID- 12902492 TI - The CD43 coreceptor molecule recruits the zeta-chain as part of its signaling pathway. AB - CD43 is an abundant cell surface sialoglycoprotein implicated in hemopoietic cell adhesion and activation. Cell stimulation through CD43 results in recruitment of different signaling proteins, including members of the Src family kinases, Syk, phospholipase Cgamma2, the adapter protein Shc, the guanine nucleotide exchange factor Vav, and activation of protein kinase C. In this study, we report that in human T lymphocytes, the zeta-chain is part of the CD43 signaling pathway. Upon CD43 engagement, the zeta-chain was tyrosine-phosphorylated, generating docking sites for tyrosine-phosphorylated zeta-associated protein of 70 kDa and Vav. In vitro kinase assays suggested that zeta-associated protein of 70 kDa could account for the kinase activity associated with the zeta-chain following CD43 engagement. Cross-linking CD43 on the surface of the Lck-deficient JCaM.1 cells failed to phosphorylate the zeta-chain and associated proteins, suggesting that Lck is a key element in the CD43 signaling pathway leading to zeta phosphorylation. CD43 engagement with beads coated with anti-CD43 mAb resulted in concentration of the zeta-chain toward the bead attachment site, but interestingly, the distribution of the T cell Ag receptor complex remained unaffected. Recruitment of the zeta-chain through CD43-mediated signals was not restricted to T lymphocytes because phosphorylation and redistribution of the zeta-chain was also observed in NK cells. Our results provide evidence that the zeta-chain functions as a scaffold molecule in the CD43 signaling pathway, favoring the recruitment and formation of downstream signaling complexes involved in the CD43-mediated cell activation of T lymphocytes and other leukocytes such as NK cells. PMID- 12902494 TI - Beta 2-microglobulin-free HLA class I heavy chain epitope mimicry by monoclonal antibody HC-10-specific peptide. AB - mAb HC-10 loses its reactivity with HLA class I (HLA-I) H chain (HC) following its association with beta(2)-microglobulin (beta(2)m). Furthermore, the HC-10 defined epitope appears to be involved in the pathogenesis of spondyloarthropathies, because HC-10 reduced their incidence in HLA B27(+)beta(2)m degrees /MHC class II knockout mice. This study has characterized the determinant recognized by HC-10. Panning of a phage display peptide library with HC-10 resulted in isolation of the motif PxxWDR, which could be aligned with P57, W60, D61, and R62 of the first domain of the HLA-I HC allospecificities reactive with HC-10. The (55)EGPEYWDR(N/E)T(64) (p-1) is the shortest motif bearing peptide that reacts with HC-10 and inhibits its binding to soluble HLA-B7 HC, irrespective of whether N (p-1a) or E (p-1b) is present at position 63. By contrast, HC-10 did not react with six additional peptides, each bearing motif amino acid substitutions present in HC-10-not-reactive HLA-I allospecificities. The p-1-derived Qp-1, synthesized with the additional conserved Q54, which displays the highest in vitro reactivity with HC-10, was the only one to induce in mice IgG resembling HC-10 in their fine specificity. Mapping of the HC-10 defined determinant suggests that the lack of mAb reactivity with beta(2)m associated HLA-I HC is caused by blocking by the peptide in the groove of beta(2)m-associated HLA-I HC, though a role of HC conformational changes following its association with beta(2)m cannot be excluded. This information contributes to our understanding of the molecular basis of the antigenic profiles of beta(2)m-free and beta(2)m-associated HLA-I HC and may serve to develop active specific immunotherapy of spondyloarthropathies. PMID- 12902495 TI - A novel -66T/C polymorphism in Fc epsilon RI alpha-chain promoter affecting the transcription activity: possible relationship to allergic diseases. AB - We found a novel polymorphism, -66T/C, in the promoter region of human FcepsilonRIalpha, the specific component of the high affinity receptor for IgE (FcepsilonRI), which is essential for the cell surface expression of FcepsilonRI and the binding of IgE Ab. When the effect of the single nucleotide replacement on the promoter function was analyzed, the transcription activity of the T allele promoter was found to be higher than that of the C allele promoter, and was markedly up-regulated by the overexpression of GATA-1 when compared with the C allele promoter. This is probably because the promoter with T at -66 has an additional GATA-1-binding motif in the region, which may assure higher affinity of the transcription factor to the promoter. In accordance with this, EMSA actually indicated that GATA-1 bound to the T allele probe (-80/-59) with the affinity higher than that to the C allele probe. Statistical analysis suggested that a significant portion of nonallergic individuals has heterozygous -66T/C genotype, while most of allergic individuals have homozygous -66T/T genotype in Japanese population. Our findings for the first time demonstrate the presence of FcepsilonRIalpha polymorphism related to the allergic diseases. PMID- 12902496 TI - Recombinant TCR ligand induces early TCR signaling and a unique pattern of downstream activation. AB - Recombinant TCR ligands (RTLs) consisting of covalently linked alpha(1) and beta(1) domains of MHC class II molecules tethered to specific antigenic peptides represent minimal TCR ligands. In a previous study we reported that the rat RTL201 construct, containing RT1.B MHC class II domains covalently coupled to the encephalitogenic guinea pig myelin basic protein (Gp-MBP(72-89)) peptide, could prevent and treat actively and passively induced experimental autoimmune encephalomyelitis in vivo by selectively inhibiting Gp-MBP(72-89) peptide specific CD4(+) T cells. To evaluate the inhibitory signaling pathway, we tested the effects of immobilized RTL201 on T cell activation of the Gp-MBP(72-89) specific A1 T cell hybridoma. Activation was exquisitely Ag-specific and could not be induced by RTL200 containing the rat MBP(72-89) peptide that differed by a threonine for serine substitution at position 80. Partial activation by RTL201 included a CD3zeta p23/p21 ratio shift, ZAP-70 phosphorylation, calcium mobilization, NFAT activation, and transient IL-2 production. In comparison, anti CD3epsilon treatment produced stronger activation of these cellular events with additional activation of NF-kappaB and extracellular signal-regulated kinases as well as long term increased IL-2 production. These results demonstrate that RTLs can bind directly to the TCR and modify T cell behavior through a partial activation mechanism, triggering specific downstream signaling events that deplete intracellular calcium stores without fully activating T cells. The resulting Ag-specific activation of the transcription factor NFAT uncoupled from the activation of NF-kappaB or extracellular signal-regulated kinases constitutes a unique downstream activation pattern that accounts for the inhibitory effects of RTL on encephalitogenic CD4(+) T cells. PMID- 12902497 TI - Human immature monocyte-derived dendritic cells express the G protein-coupled receptor GPR105 (KIAA0001, P2Y14) and increase intracellular calcium in response to its agonist, uridine diphosphoglucose. AB - Dendritic cells (DC) are essential to the initiation of an immune response due to their unique ability to take-up and process Ag, translocate to lymph nodes, and present processed Ag to naive T cells. Many chemokines, chemokine receptors and other G protein-coupled receptors (GPCRs) are implicated in these various aspects of DC biology. Through microarray analysis, we compared expression levels of chemokines, their cognate receptors, and selected GPCRs in human monocytes and in vitro monocyte-derived immature and mature DC. Hierarchical clustering of gene expression clearly distinguishes the three cell types, most notably highlighting exceptional levels of expression of the GPCR GPR105 within the immature monocyte derived DC (MDDC) gene cluster. Little or no expression was observed within the monocyte and mature MDDC cluster. Putative functionality of the GPR105 receptor was demonstrated by an observed calcium flux in immature MDDC treated with the potent GPR105 agonist, uridine 5'-diphosphoglucose (UDP-glucose), while no response to the nucleotide sugar was seen in monocytes and mature MDDC. This UDP glucose-induced calcium response was, at least in part, pertussis toxin sensitive. Moreover, immature MDDC from some donors treated with UDP-glucose exhibit an increase in expression of the costimulatory molecule CD86, which correlates with the intensity of the UDP-glucose-induced calcium flux. Together, these data demonstrate differential expression of GPR105 on immature and mature MDDC and suggest a role for the receptor and its agonist ligand in DC activation. PMID- 12902498 TI - Brugia malayi microfilariae induce cell death in human dendritic cells, inhibit their ability to make IL-12 and IL-10, and reduce their capacity to activate CD4+ T cells. AB - Parasite Ag-specific T cell unresponsiveness and diminished IFN-gamma production are immunologic hallmarks of patent infection with lymph-dwelling filarial nematodes. Although this diminished responsiveness is directed primarily against the intravascular microfilarial (MF) parasite stage and mediated in part by reduced APC function, the mechanisms involved are not fully understood. In this report, we demonstrate that human dendritic cells (DC) exposed to live MF up regulate both the cell surface and gene expression of CD54 (ICAM-1). Moreover, live MF result in a 3-fold increase in DC death compared with MF-unexposed DC, primarily due to apoptosis. Notably, microarray and real-time RT-PCR data indicate that live MF concurrently up-regulate mRNA expression of proinflammatory molecules such as IL-8, RANTES, IL-1alpha, TNF-alpha, and IL-beta in DC, the presence of which is also detected at the protein level, while inhibiting the production of IL-12 (p40 and p70) and IL-10. Soluble excretory-secretory products from live MF diminished IL-12 and IL-10 production and induced DC death, although to a lesser degree. Moreover, exposure of DC to live MF resulted in a decrease in the ability of DC to promote CD4(+) T cell production of IFN-gamma and IL-5. Our findings clearly suggest that the interaction between live MF and DC is complex but contributes to the hyporesponsiveness and parasite persistence associated with the MF(+) state in the infected human. These data further suggest that MF induce an orchestrated response in APC that leads to a diminished capacity to function appropriately, which in turn has significant consequences for CD4(+) T cells. PMID- 12902499 TI - An anti-inflammatory role for V alpha 14 NK T cells in Mycobacterium bovis bacillus Calmette-Guerin-infected mice. AB - The possible contribution of NKT cells to resistance to Mycobacterium tuberculosis infection remains unclear. In this paper we characterized the Valpha14 NKT cell population following infection with Mycobacterium bovis bacillus Calmette-Guerin (BCG). BCG infection determined an early expansion of Valpha14 NKT cells in liver, lungs, and spleen, which peaked on day 8 and was sustained until day 30. However, an NK1.1(+) Valpha14 NKT population preferentially producing IFN-gamma predominated at an early stage (day 8), which was substituted by an NK1.1(-) population preferentially producing IL-4 at later stages (day 30). Despite the fact that Valpha14 NKT cell-deficient mice eliminated BCG as did control mice, they had significantly higher numbers of granulomas in liver and lungs. Additionally, while control mice developed organized small granulomas, those in Valpha14 NKT-deficient mice had signs of caseation, large cellular infiltrates, and some multinucleated macrophages, suggesting that Valpha14 NKT cells may actually work as anti-inflammatory cells by limiting excessive lymphocyte influx and tissue pathology. In agreement, we found an increased spontaneous production and mRNA expression of TNF-alpha in liver and lungs of Valpha14 NKT-deficient mice, whose neutralization in vivo by anti-TNF-alpha mAbs consistently reduced the number of granulomas in liver and lungs. Together, our results support a regulatory role for Valpha14 NKT cells in the course of BCG infection through their ability to limit the extent of inflammatory response and point to an important role for this cell subset as a regulator of the balance between protective responses and immunopathology. PMID- 12902500 TI - T cell costimulatory molecule function determines susceptibility to infection with Pneumocystis carinii in mice. AB - Loss of T cell number and function during HIV infection or secondary to pharmacologic immunosuppression renders individuals susceptible to opportunistic infections, including Pneumocystis carinii pneumonia. Because costimulatory receptors are critical for optimal T cell function, we hypothesized that these proteins would regulate susceptibility to opportunistic infections. We found that despite normal T cell numbers, mice deficient in the costimulatory molecules CD2 and CD28 spontaneously developed P. carinii pneumonia. In experiments using intratracheal injection of P. carinii organisms to induce infection, the loss of CD28 alone was sufficient to render mice susceptible to acute infection; however, the organism was eventually cleared. Examination of inflammatory responses to P. carinii revealed that mice deficient in both CD2 and CD28 accumulated CD8(+) T cells in their lungs in response to infection and demonstrated markedly reduced specific Ab titers. Analysis of cytokine profiles suggested that regulation of IL 10 and IL-15 may be important elements of the response to this pathogen. Thus, costimulatory molecule function is critical in determining the initial susceptibility to infection with P. carinii. Analysis of immunologic responses in these mice may provide important insights into the defects that render individuals susceptible to opportunistic infection, and provide opportunities for novel immunologically based therapies. PMID- 12902501 TI - The fourth surface-exposed region of the outer membrane protein P5-homologous adhesin of nontypable Haemophilus influenzae is an immunodominant but nonprotective decoying epitope. AB - Nontypable Haemophilus influenzae is a major cause of otitis media and other mucosal infections. After natural disease in children and experimental disease in chinchillas, we found a hierarchical pattern of immunodominance among the four surface-exposed regions of the P5-homologous adhesin, with the greatest response directed to region 4. However, Ab to region 4 is not protective. When this natural but biased response was refocused to region 3 by immunization, augmented bacterial clearance and protection from ascending otitis media was observed. Collectively, the data indicate that region 4 contains a highly immunodominant but nonprotective decoying epitope, the presence of which dampens the immune response to a subdominant but protective epitope in region 3. PMID- 12902502 TI - Presentation and binding affinity of equine infectious anemia virus CTL envelope and matrix protein epitopes by an expressed equine classical MHC class I molecule. AB - Control of a naturally occurring lentivirus, equine infectious anemia virus (EIAV), occurs in most infected horses and involves MHC class I-restricted, virus specific CTL. Two minimal 12-aa epitopes, Env-RW12 and Gag-GW12, were evaluated for presentation by target cells from horses with an equine lymphocyte Ag-A1 (ELA A1) haplotype. Fifteen of 15 presented Env-RW12 to CTL, whereas 11 of 15 presented Gag-GW12. To determine whether these epitopes were presented by different molecules, MHC class I genes were identified in cDNA clones from Arabian horse A2152, which presented both epitopes. This horse was selected because it is heterozygous for the SCID trait and is used to breed heterozygous females. Offspring with SCID are used as recipients for CTL adoptive transfer, and normal offspring are used for CTL induction. Four classical and three putative nonclassical full-length MHC class I genes were found. Human 721.221 cells transduced with retroviral vectors expressing each gene had equine MHC class I on their surface. Following peptide pulsing, only cells expressing classical MHC class I molecule 7-6 presented Env-RW12 and Gag-GW12 to CTL. Unlabeled peptide inhibition of (125)I-labeled Env-RW12 binding to 7-6-transduced cells demonstrated that Env-RW12 affinity was 15-fold higher than Gag-GW12 affinity. Inhibition with truncated Env-RW12 demonstrated that amino acid positions 1 and 12 were necessary for binding, and single substitutions identified positions 2 and 3 as possible primary anchor residues. Since MHC class I 7-6 presented both epitopes, outbred horses with this allele can be immunized with these epitopes to optimize CTL responses and evaluate their effectiveness against lentiviral challenge. PMID- 12902503 TI - Nramp1 functionality increases inducible nitric oxide synthase transcription via stimulation of IFN regulatory factor 1 expression. AB - Natural-resistance associated macrophage protein 1 (Nramp1) encodes a transmembrane phagolysosomal protein exerting resistance toward infections with intracellular pathogens by a mechanism not fully elucidated so far. We used the murine macrophage cell line RAW264.7, stably transfected with functional (RAW-37) or nonfunctional (RAW-21) Nramp1, to study for differences in the expression of NO, a central antimicrobial effector molecule of macrophages. Following stimulation with IFN-gamma and LPS, Nramp1-expressing cells exhibit higher enzymatic activity of inducible NO synthase (iNOS) and increased cytoplasmic iNOS mRNA levels than RAW-21 cells. Time-course experiments showed that iNOS-mRNA levels remain increased in RAW-37 cells after prolonged cytokine stimulation while they decrease in RAW-21 cells. Reporter gene assays with iNOS-promoter luciferase constructs demonstrated an increased and prolonged promoter activity in Nramp1-resistant vs susceptible cells. This was paralleled by increased IFN regulatory factor 1 (IRF-1) expression and binding affinity to the iNOS promoter in RAW-37 cells, which may be related to enhanced STAT-1 binding affinity in these cells. A point mutation within the IRF-1 binding site of the iNOS promoter abolished the differences in iNOS transcription between RAW-21 and RAW-37 cells. Cells carrying functional Nramp1 express increased amounts of NO, which may be related to STAT-1-mediated stimulation of IRF-1 expression with subsequent prolonged activation of iNOS transcription. Enhanced NO expression may partly underlie the protection against infection with intracellular pathogens by Nramp1 functionality. PMID- 12902504 TI - DNA vaccines, combining form of antigen and method of delivery to raise a spectrum of IFN-gamma and IL-4-producing CD4+ and CD8+ T cells. AB - DNA-based immunizations have been used to determine the patterns of type 1 and type 2 cytokines that can be induced in vivo for Ag-specific CD4(+) and CD8(+) T cells. IL-4 was used as a signature cytokine for a type 2 T cell response and IFN gamma as the signature cytokine for a type 1 response. Gene gun deliveries of secreted Ags were used to bias responses toward type 2 and saline injections of cell-associated Ags to bias responses toward type 1. The studies revealed that gene gun bombardments of DNAs expressing secreted Ags strongly biased responses toward type 2, inducing IL-4-producing CD8(+) as well as CD4(+) T cells. Saline injections of DNAs expressing cell-associated Ags strongly biased responses toward type 1, inducing IFN-gamma-producing CD8(+) and CD4(+) cells. A mixed type 1/type 2 response of IFN-gamma-producing CD8(+) T cells and IL-4-producing CD4(+) T cells was found for gene gun deliveries of cell-associated Ags. Saline injections of secreted Ags raised a weakly type 1-biased response characterized by only slightly higher frequencies of IFN-gamma- than IL-4-producing CD4(+) and CD8(+) T cells. Studies in B cell knockout and hen egg lysozyme Ig transgenic mice revealed that B cells were required for the generation of IL-4-producing CD8(+) T cells. PMID- 12902505 TI - Protection against CTL escape and clinical disease in a murine model of virus persistence. AB - CTL escape mutations have been identified in several chronic infections, including mice infected with mouse hepatitis virus strain JHM. One outstanding question in understanding CTL escape is whether a CD8 T cell response to two or more immunodominant CTL epitopes would prevent CTL escape. Although CTL escape at multiple epitopes seems intuitively unlikely, CTL escape at multiple CD8 T cell epitopes has been documented in some chronically infected individual animals. To resolve this apparent contradiction, we engineered a recombinant variant of JHM that expressed the well-characterized gp33 epitope of lymphocytic choriomeningitis virus, an epitope with high functional avidity. The results show that the presence of a host response to this second epitope protected mice against CTL escape at the immunodominant JHM-specific CD8 T cell epitope, the persistence of infectious virus, and the development of clinical disease. PMID- 12902506 TI - Lipomannans, but not lipoarabinomannans, purified from Mycobacterium chelonae and Mycobacterium kansasii induce TNF-alpha and IL-8 secretion by a CD14-toll-like receptor 2-dependent mechanism. AB - Lipoarabinomannans (LAMs) are glycolipids from the mycobacterial cell wall that exhibit various biological activities, including proinflammatory and anti inflammatory responses. However, little is known about the properties of lipomannans (LMs), considered to be precursors of LAMs. In this study, we provide evidence that LMs purified from Mycobacterium chelonae and a clinical strain of Mycobacterium kansasii stimulated mRNA expression and secretion of TNF-alpha and IL-8 from human macrophage-like differentiated THP-1 cells. In contrast to LMs, LAMs were not able to induce a significant cytokine-inducing effect. The mechanism of activation by LMs was investigated using various Abs raised against surface receptors for multiple bacterial products. The presence of anti-CD14 or anti-Toll-like receptor 2 (TLR2) Abs profoundly affected production of TNF-alpha and IL-8, suggesting that both CD14 and TLR2 participate in the LM-mediated activation process. Furthermore, stimulation of cells was dependent on the presence of the LPS-binding protein, a plasma protein that transfers glycolipids to CD14. Chemical degradation of the arabinan domain of mannose-capped LAM from M. kansasii, which presented no cytokine-eliciting effect, restored the cytokine inducing activity at a level similar to those of LMs. These results support the hypothesis that the presence of an arabinan in LAMs prevents the interaction of these glycolipids with TLR2/CD14 receptors. In addition, we found that phosphatidylinositol dimannosides isolated from M. kansasii did not induce cytokine secretion. This study suggests that LMs isolated from different mycobacterial species participate in the immunomodulation of the infected host and that the D-mannan core of this glycolipid is essential for this function. PMID- 12902507 TI - Protracted protection to Plasmodium berghei malaria is linked to functionally and phenotypically heterogeneous liver memory CD8+ T cells. AB - We previously demonstrated that protection induced by radiation-attenuated (gamma) Plasmodium berghei sporozoites is linked to MHC class I-restricted CD8(+) T cells specific for exoerythrocytic-stage Ags, and that activated intrahepatic memory CD8(+) T cells are associated with protracted protection. In this study, we further investigated intrahepatic memory CD8(+) T cells to elucidate mechanisms required for their maintenance. Using phenotypic markers indicative of activation (CD44, CD45RB), migration (CD62L), and IFN-gamma production, we identified two subsets of intrahepatic memory CD8(+) T cells: the CD44(high)CD45RB(low)CD62L(low)CD122(low) phenotype, representing the dominant effector memory set, and the CD44(high)CD45RB(high)CD62L(low/high)CD122(high) phenotype, representing the central memory set. Only the effector memory CD8(+) T cells responded swiftly to sporozoite challenge by producing sustained IFN-gamma; the central memory T cells responded with delay, and the IFN-gamma reactivity was short-lived. In addition, the subsets of liver memory CD8(+) T cells segregated according to the expression of CD122 (IL-15R) in that only the central memory CD8(+) T cells were CD122(high), whereas the effector memory CD8(+) T cells were CD122(low). Moreover, the effector memory CD8(+) T cells declined as protection waned in mice treated with primaquine, a drug that interferes with the formation of liver-stage Ags. We propose that protracted protection induced by P. berghei radiation-attenuated sporozoites depends in part on a network of interactive liver memory CD8(+) T cell subsets, each representing a different phase of activation or differentiation, and the balance of which is profoundly affected by the repository of liver-stage Ag and IL-15. PMID- 12902508 TI - Helicobacter pylori infection interferes with epithelial Stat6-mediated interleukin-4 signal transduction independent of cagA, cagE, or VacA. AB - Helicobacter pylori is a bacterial pathogen evolved to chronically colonize the gastric epithelium, evade immune clearance by the host, and cause gastritis, peptic ulcers, and even gastric malignancies in some infected humans. In view of the known ability of this bacterium to manipulate gastric epithelial cell signal transduction cascades, we determined the effects of H. pylori infection on epithelial IL-4-Stat6 signal transduction. HEp-2 and MKN45 epithelial cells were infected with H. pylori strains LC11 or 8823 (type 1; cagA(+)/cagE(+)/VacA(+)), LC20 (type 2; cagA(-), cagE(-), VacA(-)), and cagA, cagE, and vacA isogenic mutants of strain 8823, with some cells receiving subsequent treatment with the Th2 cytokine IL-4, a known Stat6 activator. Immunofluorescence showed a disruption of Stat6-induced nuclear translocation by IL-4 in LC11-infected HEp-2 cells. IL-4-inducible Stat6 DNA binding in HEp-2 and MKN45 cells was abrogated by infection, but MKN45 cell viability was unaffected. A decrease in IL-4-mediated Stat6 tyrosine phosphorylation in nuclear and whole cell lysates was also observed following infection with strains LC11 and LC20, while neither strain altered IL-4 receptor chain alpha or Janus kinase 1 protein expression. Furthermore, parental strain 8823 and its isogenic cagA, cagE, and vacA mutants also suppressed IL-4-induced Stat6 tyrosine phosphorylation to comparable degrees. Thus, H. pylori did not directly activate Stat6, but blocked the IL-4 induced activation of epithelial Stat6. This may represent an evolutionarily conserved strategy to disrupt a Th2 response and evade the host immune system, allowing for successful chronic infection. PMID- 12902509 TI - Mycobacterium tuberculosis exerts gene-selective inhibition of transcriptional responses to IFN-gamma without inhibiting STAT1 function. AB - Mycobacterium tuberculosis is a highly successful human pathogen. A major component of this success is the pathogen's ability to avoid eradication by the innate and adaptive immune responses throughout the course of infection. IFN gamma, a potent activator of the microbicidal activities of macrophages, is essential for control of M. tuberculosis infection, but is unable to stimulate macrophages to kill M. tuberculosis. We have found that infection of the human monocytic cell line, THP-1, resulted in reduced cellular responses to IFN-gamma, manifested as impaired induction of CD64 surface expression and transcription. This defect in transcription occurred despite normal activation of STAT1 in infected macrophages: there was no decrease in STAT1 tyrosine or serine phosphorylation, nuclear translocation, or binding of a minimal IFN-gamma response sequence. Assays of STAT1 function in M. tuberculosis-treated cells also revealed no defect in activation of a minimal gamma-activated sequence construct or STAT1 recruitment to and binding at the endogenous CD64 promoter. In addition, M. tuberculosis did not affect histone acetylation at the CD64 promoter. The inhibition of transcription was gene selective: while transcription of CD64 and class II transactivator were decreased, certain other IFN-gamma-responsive genes either were unaffected or were increased by M. tuberculosis. These results indicate that M. tuberculosis inhibits the response to IFN-gamma by a mechanism distinct from either suppressor of cytokine signaling-1 inhibition of STAT1 phosphorylation or protein inhibitor of activated STAT interference with DNA binding, and indicate that other mechanisms of inhibition of IFN-gamma responses remain to be discovered. PMID- 12902510 TI - Expression and function of formyl peptide receptors on human fibroblast cells. AB - The migration of polymorphonuclear leukocytes from the blood to sites of infection in tissues is a hallmark of the innate immune response. Formylated peptides produced as a byproduct of bacterial protein synthesis are powerful chemoattractants for leukocytes. Formyl peptides bind to two different G protein coupled receptors (formyl peptide receptor (FPR) and the low affinity formyl peptide receptor-like-1 (FPRL1)) to initiate a signal transduction cascade leading to cell activation and migration. Our analysis of expressed sequences from many cDNA libraries draws attention to the fact that FPRs are widely expressed in nonlymphoid tissues. Here we demonstrate that FPRs are expressed by normal human lung and skin fibroblasts and the human fibrosarcoma cell line HT 1080. The expression on fibroblasts of receptors for bacteria-derived peptides raises questions about the possible function of these receptors in nonleukocyte cells. We studied the function of FPRs on fibroblasts and find that stimulation with fMLP triggers dose-dependent migration of these cells. Furthermore, fMLP induces signal transduction including intracellular calcium flux and a transient increase in F-actin. The fMLP-induced adhesion and motility of fibroblasts on fibronectin require functional protein kinase C and phosphatidylinositol 3 kinase. This first report of a functional formyl peptide receptor in cells of fibroblast origin opens new possibilities for the role of fibroblasts in innate immune responses. PMID- 12902511 TI - Plasminogen activator inhibitor-1 supports IL-8-mediated neutrophil transendothelial migration by inhibition of the constitutive shedding of endothelial IL-8/heparan sulfate/syndecan-1 complexes. AB - The endothelium is the primary barrier to leukocyte recruitment at sites of inflammation. Neutrophil recruitment is directed by transendothelial gradients of IL-8 that, in vivo, are bound to the endothelial cell surface. We have investigated the identity and function of the binding site(s) in an in vitro model of neutrophil transendothelial migration. In endothelial culture supernatants, IL-8 was detected in a trimolecular complex with heparan sulfate and syndecan-1. Constitutive shedding of IL-8 in this form was increased in the presence of a neutralizing Ab to plasminogen activator inhibitor-1 (PAI-1), indicating a role for endothelial plasminogen activator in the shedding of IL-8. Increased shedding of IL-8/heparan sulfate/syndecan-1 complexes was accompanied by inhibition of neutrophil transendothelial migration, and aprotinin, a potent plasmin inhibitor, reversed this inhibition. Platelets, added as an exogenous source of PAI-1, had no effect on shedding of the complexes or neutrophil migration. Our results indicate that IL-8 is immobilized on the endothelial cell surface through binding to syndecan-1 ectodomains, and that plasmin, generated by endothelial plasminogen activator, induces the shedding of this form of IL-8. PAI 1 appears to stabilize the chemoattractant form of IL-8 at the cell surface and may represent a therapeutic target for novel anti-inflammatory strategies. PMID- 12902512 TI - Inflammatory chemoreceptor cross-talk suppresses leukotriene B4 receptor 1 mediated neutrophil calcium mobilization and chemotaxis after trauma. AB - G protein-coupled chemoattractants recruit neutrophils (PMN) to sites of injury and infection. The leukotrienes (LT) and CXC chemokines (CXC) and their receptors (BLT1/BLT2 and CXCR1/CXCR2) are all known to play roles in these responses. Each system has been studied separately in vitro, but in vivo they act concurrently, and the clinical interactions between the two systems are unstudied. We prospectively studied calcium mobilization and chemotactic responses to LTB(4) in PMN from major trauma patients. The responses of the high affinity BLT1 receptor were suppressed at the 3-day postinjury time point, but recovered by 1 wk. Trauma patients had transient elevations of plasma LT and CXC levels. Functional deficits identical with those in trauma PMN were reproduced in vitro by exposing healthy PMN to CXCs at the elevated plasma concentrations found. Functional responses to LTB(4) were suppressed by cross-talk with CXC and BLT2 receptors that desensitize BLT1. Since the suppression of intracellular calcium mobilization was prominent, we also studied the role of suppressed cell calcium mobilization in the defective chemotactic responses to LTB(4). We noted that PMN chemotaxis to LTB(4) showed far more dependence on store-operated calcium entry than on the release of cellular calcium stores, and that store-operated calcium responses to BLT1 activation were markedly inhibited during the same time period as was chemotaxis. The intermittent release of inflammatory mediators after injury can blunt PMN responses to LTs by suppressing BLT1 as well as downstream calcium entry. Diminished LT receptor activity due to cross-talk with CXC receptors can inhibit PMN recruitment to infective sites. This may predispose injured patients to septic complications. PMID- 12902513 TI - Functional expression of neurokinin 1 receptors on mast cells induced by IL-4 and stem cell factor. AB - It is widely accepted that neurokinin 1 (NK(1)) receptors are not generally expressed on mast cells but little is known about their expression in inflammation. The present study shows expression of NK(1) receptors on bone marrow-derived mast cells (BMMC) under the influence of IL-4 or stem cell factor (SCF). Highest expression was found when both cytokines are present. Six days of coculture with the cytokines IL-4 and SCF showed significant expression of NK(1) receptors (NK(1) receptor(+)/c-kit(+) BMMC; control: 7%, IL-4/SCF: 16%), while 12 days of cytokine coculture increased this expression to 37% positive cells. A longer coculture with IL-4 and SCF did not give an additional effect. Increased expression in IL-4/SCF-treated BMMC was further confirmed using Western blot analysis. Next, we demonstrated the functional relevance of NK(1) receptor expression for mast cell activation, resulting in an enhanced degranulation upon stimulation by substance P. BMMC activation was significantly diminished by the NK(1) receptor antagonist RP67580 (10 micro M) when stimulated with low concentrations of substance P. The inactive enantiomer RP65681 had no effect. In addition, BMMC cultured from bone marrow of NK(1) receptor knockout mice showed significantly decreased exocytosis to low concentrations of substance P. The present study clearly shows that NK(1) receptor-induced activation contributes significantly at low physiological substance P concentrations (<100 micro M). In conclusion, BMMC were shown to express NK(1) receptors upon IL-4/SCF coculture. This expression of NK(1) receptors has been demonstrated to be of functional relevance and leads to an increase in the sensitivity of BMMC to substance P. PMID- 12902514 TI - Airway IgG counteracts specific and bystander allergen-triggered pulmonary inflammation by a mechanism dependent on Fc gamma R and IFN-gamma. AB - Besides IgE, the Ab isotype that gives rise to sensitization and allergic asthma, the immune response to common inhalant allergens also includes IgG. Increased serum titers of allergen-specific IgG, induced spontaneously or by allergen vaccination, have been implicated in protection against asthma. To verify the interference of topical IgG with the allergen-triggered eosinophilic airway inflammation that underlies asthma, sensitized mice were treated by intranasal instillation of specific IgG, followed by allergen challenge. This treatment strongly reduced eosinophilic inflammation and goblet cell metaplasia, and increased Th1 reactivity and IFN-gamma levels in bronchoalveolar lavage fluid. In contrast, inflammatory responses were unaffected in IFN-gamma-deficient mice or when applying F(ab')(2). Although dependent on specific allergen-IgG interaction, inflammation triggered by bystander allergens was similarly repressed. Perseverance of inflammation repression, apparent after secondary allergen challenge, and increased allergen capture by alveolar macrophages further characterized the consequences of topical IgG application. These results assign a novel protective function to anti-allergen IgG namely at the local level interference with the inflammatory cascade, resulting in repression of allergic inflammation through an FcgammaR- and IFN-gamma-dependent mechanism. Furthermore, these results provide a basis for topical immunotherapy of asthma by direct delivery of anti-allergen IgG to the airways. PMID- 12902515 TI - Synergism between platelet-activating factor-like phospholipids and peroxisome proliferator-activated receptor gamma agonists generated during low density lipoprotein oxidation that induces lipid body formation in leukocytes. AB - Oxidized low density lipoprotein (LDL) has an important proinflammatory role in atherogenesis. In this study, we investigated the ability of oxidized LDL (oxLDL) and its phospholipid components to induce lipid body formation in leukocytes. Incubation of mouse peritoneal macrophages with oxidized, but not with native LDL led to lipid body formation within 1 h. This was blocked by platelet-activating factor (PAF) receptor antagonists or by preincubation of oxLDL with rPAF acetylhydrolase. HPLC fractions of phospholipids purified from oxLDL induced calcium flux in neutrophils as well as lipid body formation in macrophages. Injection of the bioactive phospholipid fractions or butanoyl and butenoyl PAF, a phospholipid previously shown to be present in oxLDL, into the pleural cavity of mice induced lipid body formation in leukocytes recovered after 3 h. The 5 lipoxygenase and cyclooxygenase-2 colocalized within lipid bodies formed after stimulation with oxLDL, bioactive phospholipid fractions, or butanoyl and butenoyl PAF. Lipid body formation was inhibited by 5-lipoxygenase antagonists, but not by cyclooxygenase-2 inhibitors. Azelaoyl-phosphatidylcholine, a peroxisome proliferator-activated receptor-gamma agonist in oxLDL phospholipid fractions, induced formation of lipid bodies at late time points (6 h) and synergized with suboptimal concentrations of oxLDL. We conclude that lipid body formation is an important proinflammatory effect of oxLDL and that PAF-like phospholipids and peroxisome proliferator-activated receptor-gamma agonists generated during LDL oxidation are important mediators in this phenomenon. PMID- 12902517 TI - Prevention of collagen-induced arthritis in mice transgenic for the complement inhibitor complement receptor 1-related gene/protein y. AB - The objective of these studies was to examine collagen-induced arthritis (CIA) in C57BL/6 mice transgenic for the rodent complement regulatory protein complement receptor 1-related gene/protein y (Crry) (Crry-Tg), a C3 convertase inhibitor. The scores for clinical disease activity and for histological damage in the joints were both significantly decreased in Crry-Tg mice in comparison to wild type (WT) littermates. The production of both IgG1 and IgG2a anti-collagen Abs was reduced in the Crry-Tg mice, although spleen cell proliferation in response to collagen type II was not altered. The production of IFN-gamma, TNF-alpha, and IL-1beta by LPS-stimulated spleen cells was decreased, and IL-10 was increased, in cells from Crry-Tg mice in comparison to WT. The steady-state mRNA levels for IFN-gamma, TNF-alpha, and IL-1beta were all decreased in the joints of Crry-Tg mice in comparison to WT. The synovium from Crry-Tg mice without CIA contained the mRNA for the Crry transgene, by RT-PCR, and the synovium from transgenic mice with CIA exhibited little deposition of C3 protein by immunohistological analysis. These results suggest that suppression of CIA in Crry-Tg mice may be due to enhanced synthesis of Crry locally in the joint with decreased production of proinflammatory cytokines. PMID- 12902516 TI - Intracellular domain of brain endothelial intercellular adhesion molecule-1 is essential for T lymphocyte-mediated signaling and migration. AB - To examine the role of the ICAM-1 C-terminal domain in transendothelial T lymphocyte migration and ICAM-1-mediated signal transduction, mutant human (h)ICAM-1 molecules were expressed in rat brain microvascular endothelial cells. The expression of wild-type hICAM-1 resulted in a significant increase over basal levels in both adhesion and transendothelial migration of T lymphocytes. Endothelial cells (EC) expressing ICAM-1 in which the tyrosine residue at codon 512 was substituted with phenylalanine (hICAM-1(Y512F)) also exhibited increased lymphocyte migration, albeit less than that with wild-type hICAM-1. Conversely, the expression of truncated hICAM-1 proteins, in which either the intracellular domain was deleted (hICAM-1DeltaC) or both the intracellular and transmembrane domains were deleted through construction of a GPI anchor (GPI-hICAM-1), did not result in an increase in lymphocyte adhesion, and their ability to increase transendothelial migration was attenuated. Truncated hICAM-1 proteins were also unable to induce ICAM-1-mediated Rho GTPase activation. EC treated with cell permeant penetratin-ICAM-1 peptides comprising human or rat ICAM-1 intracellular domain sequences inhibited transendothelial lymphocyte migration, but not adhesion. Peptides containing a phosphotyrosine residue were equipotent in inhibiting lymphocyte migration. These data demonstrate that the intracellular domain of ICAM-1 is essential for transendothelial migration of lymphocytes, and that peptidomimetics of the ICAM-1 intracellular domain can also inhibit this process. Such competitive inhibition of transendothelial lymphocyte migration in the absence of an affect on adhesion further implicates ICAM-1-mediated signaling events in the facilitation of T lymphocyte migration across brain EC. Thus, agents that mimic the ICAM-1 intracellular domain may be attractive targets for novel anti-inflammatory therapeutics. PMID- 12902518 TI - A plant-based allergy vaccine suppresses experimental asthma via an IFN-gamma and CD4+CD45RBlow T cell-dependent mechanism. AB - Allergic asthma is currently considered a chronic airway inflammatory disorder associated with the presence of activated CD4(+) Th2-type lymphocytes, eosinophils, and mast cells. Interestingly, therapeutic strategies based on immune deviation and suppression have been shown to successfully attenuate the development of the asthma phenotype. In this investigation, we have for the first time used a genetically modified (GM) plant, narrow leaf lupin (Lupinus angustifolius L.), expressing a gene for a potential allergen (sunflower seed albumin) (SSA-lupin) to examine whether a GM plant/food-based vaccine strategy can be used to suppress the development of experimental asthma. We show that oral consumption of SSA-lupin promoted the induction of an Ag-specific IgG2a Ab response. Furthermore, we demonstrate that the plant-based vaccine attenuated the induction of delayed-type hypersensitivity responses and pathological features of experimental asthma (mucus hypersecretion, eosinophilic inflammation, and enhanced bronchial reactivity (airways hyperreactivity). The suppression of experimental asthma by SSA-lupin was associated with the production of CD4(+) T cell-derived IFN-gamma and IL-10. Furthermore, we show that the specific inhibition of experimental asthma was mediated via CD4(+)CD45RB(low) regulatory T cells and IFN-gamma. Thus, our data demonstrate that a GM plant-based vaccine can promote a protective immune response and attenuate experimental asthma, suggesting that plant-based vaccines may be potentially therapeutic for the protection against allergic diseases. PMID- 12902519 TI - A novel therapeutic approach targeting articular inflammation using the filarial nematode-derived phosphorylcholine-containing glycoprotein ES-62. AB - Understanding modulation of the host immune system by pathogens offers rich therapeutic potential. Parasitic filarial nematodes are often tolerated in human hosts for decades with little evidence of pathology and this appears to reflect parasite-induced suppression of host proinflammatory immune responses. Consistent with this, we have previously described a filarial nematode-derived, secreted phosphorylcholine-containing glycoprotein, ES-62, with immunomodulatory activities that are broadly anti-inflammatory in nature. We sought to evaluate the therapeutic potential of ES-62 in vitro and in vivo in an autoimmune disease model, namely, collagen-induced arthritis in DBA/1 mice. ES-62 given during collagen priming significantly reduced initiation of inflammatory arthritis. Crucially, ES-62 was also found to suppress collagen-induced arthritis severity and progression when administration was delayed until after clinically evident disease onset. Ex vivo analyses revealed that in both cases, the effects were associated with inhibition of collagen-specific pro-inflammatory/Th1 cytokine (TNF-alpha, IL-6, and IFN-gamma) release. In parallel in vitro human tissue studies, ES-62 was found to significantly suppress macrophage activation via cognate interaction with activated T cells. Finally, ES-62 suppressed LPS-induced rheumatoid arthritis synovial TNF-alpha and IL-6 production. Evolutionary pressure has promoted the generation by pathogens of diverse mechanisms enabling host immune system evasion and induction of "tolerance." ES-62 represents one such mechanism. We now provide proof of concept that parasite-derived immunomodulatory strategies offer a novel therapeutic opportunity in inflammatory arthritis. PMID- 12902520 TI - Differentiation of CD8+ T cells from tumor-invaded and tumor-free lymph nodes of melanoma patients: role of common gamma-chain cytokines. AB - Differentiation of CD8(+) T cells at the tumor site toward effector and memory stages may represent a key step for the efficacy of antitumor response developing naturally or induced through immunotherapy. To address this issue, CD8(+) T lymphocytes from tumor-invaded (n = 142) and tumor-free (n = 42) lymph nodes removed from the same nodal basin of melanoma patients were analyzed for the expression of CCR7, CD45RA, perforin, and granzyme B. By hierarchical cluster analysis, CD8(+) T cells from all tumor-free lymph nodes and from 56% of the tumor-invaded lymph node samples fell in the same cluster, characterized mainly by CCR7(+) CD45RA(+/-) cytotoxic factor(-) cells. The remaining three clusters contained only samples from tumor-invaded lymph nodes and showed a progressive shift of the CD8(+) T cell population toward CCR7(-) CD45RA(-/+) perforin(+) granzyme B(+) differentiation stages. Distinct CD8(+) T cell maturation stages, as defined by CCR7 vs CD45RA and by functional assays, were identified even in melanoma- or viral Ag-specific T cells from invaded lymph nodes by HLA tetramer analysis. Culture for 7 days of CCR7(+) perforin(-) CD8(+) T cells from tumor invaded lymph nodes with IL-2 or IL-15, but not IL-7, promoted, mainly in CCR7(+)CD45RA(-) cells, proliferation coupled to differentiation to the CCR7(-) perforin(+) stage and acquisition of melanoma Ag-specific effector functions. Taken together, these results indicate that CD8(+) T cells differentiated toward CCR7(-) cytotoxic factor(+) stages are present in tumor-invaded, but not in tumor free, lymph nodes of a relevant fraction of melanoma patients and suggest that cytokines such as IL-2 and IL-15 may be exploited to promote Ag-independent maturation of anti-tumor CD8(+) T cells. PMID- 12902522 TI - Reconstitution of CD8+ T cells by retroviral transfer of the TCR alpha beta-chain genes isolated from a clonally expanded P815-infiltrating lymphocyte. AB - Gene transfer of TCR alphabeta-chains into T cells may be a promising strategy for providing valuable T lymphocytes in the treatment of tumors and other immune mediated disorders. We report in this study the reconstitution of CD8(+) T cells by transfer of TCR alphabeta-chain genes derived from an infiltrating T cell into P815. Analysis of the clonal expansion and Vbeta subfamily usage of CD8(+) TIL in the tumor sites demonstrated that T cells using Vbeta10 efficiently infiltrated and expanded clonally. The TCR alpha- and beta-chain sequences derived from a tumor-infiltrating CD8(+)/Vbeta10(+) single T cell clone (P09-2C clone) were simultaneously determined by the RT-PCR/single-strand conformational polymorphism method and the single-cell PCR method. When P09-2C TCR alphabeta-chain genes were retrovirally introduced into CD8(+) T cells, the reconstituted T cells positively lysed the P815 tumor cells, but not the A20, EL4, or YAC-1 cells, in vitro. In addition, the CTL activity was blocked by the anti-H2L(d) mAb. Furthermore, T cells containing both TCR alpha- and beta-chains, but not TCR beta-chain alone, accumulated at the tumor-inoculated site when the reconstituted CD8(+) T cells were adoptively transferred to tumor-bearing nude mice. These findings suggest that it is possible to reconstitute functional tumor-specific CD8(+) T cells by transfer of TCR alphabeta-chain genes derived from TIL, and that such T cells might be useful as cytotoxic effector cells or as a vehicle for delivering therapeutic agents. PMID- 12902521 TI - Immunoregulatory role of CD1d in the hydrocarbon oil-induced model of lupus nephritis. AB - Systemic lupus erythematosus (SLE) is a systemic autoimmune disease that is accompanied by the emergence of autoreactive T cells and a reduction in regulatory T cells. Humans and mice with SLE have reduced numbers of CD1d restricted NK T cells, suggesting a role for these cells in the regulation of SLE. In this study, we show that CD1d deficiency exacerbates lupus nephritis induced by the hydrocarbon oil pristane. This exacerbation in disease is associated with: 1) reduced TNF-alpha and IL-4 production by T cells, especially during the disease induction phase; and 2) expansion of marginal zone B cells. Strikingly, inoculation of pristane in wild-type mice resulted in reduced numbers and/or functions of NK T cells and CD1d-expressing dendritic cells. These findings suggest that CD1d may play an immunoregulatory role in the development of lupus in the pristane-induced model. PMID- 12902523 TI - HER-2/neu-specific monoclonal antibodies collaborate with HER-2/neu-targeted granulocyte macrophage colony-stimulating factor secreting whole cell vaccination to augment CD8+ T cell effector function and tumor-free survival in Her-2/neu transgenic mice. AB - HER-2/neu is overexpressed in several cancers including 30% of breast carcinomas, and correlates with a poor outcome. HER-2/neu-transgenic (neu-N) mice that overexpress the non-transforming rat neu develop spontaneous mammary carcinomas and demonstrate immunotolerance to the neu protein similar to that observed in patients with neu-expressing cancers. In neu-N mice, neu-targeted vaccination induces weak T cell and negligible Ab responses sufficient to delay but not eradicate transplanted neu-expressing tumor. Here we demonstrate that passive infusion of neu-specific mAbs in sequence with whole cell vaccination significantly improves tumor-free survival over either modality alone. Importantly, treatment of neu-N mice with vaccine in combination with two distinct neu-specific Abs is particularly efficacious, preventing tumor in 70% and eradicating established tumor in 30% of neu-N mice. In vivo lymphocyte subpopulation depletion experiments demonstrate that the efficacy of Ab, alone or combined with vaccine, is dependent on both CD4(+) and CD8(+) T cells. Furthermore, the in vivo antitumor effects of vaccine and Ab are associated with a significant increase in the number and function of neu-specific CD8(+) T cells. Collectively, these observations suggest that similarly increased efficacy could be obtained by combining neu-targeted vaccination and neu-specific Abs such as trastuzumab (Herceptin) in patients with neu-expressing cancers. PMID- 12902525 TI - Regional variation and changes with ageing in vibrotactile sensitivity in the human footsole. AB - Recently there has been indirect evidence suggesting that age-related elevation in footsole vibration detection may be associated with balance and gait dysfunction. As a first step in investigating this dysfunction, the current study determined by how much plantar vibration sensation decreases as a function of age, and if change is dependent on frequency and location of vibration application. Vibration thresholds were assessed at 4 frequencies (25-400 Hz), at 55 locations, and in young and older participants. Results showed there were 3 regions of sensitivity on the footsole: the ball/medial arch, the lateral border of the foot and heel, and the toes. Thresholds for fast-adapting type I receptor (FAI)-mediated frequencies were age invariant; however, thresholds for fast adapting type II receptor (FAII)-mediated frequencies increased with age. These changes may be one of many factors contributing to age-related changes in gait. PMID- 12902526 TI - Denervation stimulates apoptosis but not Id2 expression in hindlimb muscles of aged rats. AB - Inhibitors of differentiation (Id) proteins are repressors of myogenic regulatory factors and have been implicated in apoptosis and muscle atrophy during aging. Indeed, we have previously found that Id levels are elevated in muscles from old rodents, possibly as a consequence of loss of alpha-motoneurons during senescence. To determine if Id2 proteins increase after denervation and if this is accompanied by increased apoptosis in aged as compared with adult animals, the gastrocnemius and soleus muscles were denervated in 1 limb of Fischer 344 x Brown Norway rats aged 9 months (adult, n = 12) and 33 months (aged, n = 9), while the contralateral limb served as the intra-animal control. After 14 days, the muscles in each limb were removed. The levels of Id1, Id2, and Id3 mRNA and protein were significantly greater in muscles of old as compared with young adult rats. Denervation, however, did not significantly increase Id1, Id2, and Id3 mRNA in soleus or gastrocnemius muscles from either young or old rats. Also Id2 protein levels were similar in denervated and control muscles from young adult and old rats. In young adult rats only, denervation induced an increase in Id1 and Id3 protein levels in both the soleus (Id1 113%; Id3 900%) and gastrocnemius (Id1 86%; Id3 80%). Denervation induced a significant increase in caspase 8 in both soleus and gastrocnemius muscles from young (101% and 147%, respectively) and old rats (167% and 190%, respectively). Bax protein levels, as estimated by western blots, increased by 726% and 1087% after denervation in the soleus and by 368% and 49% in the gastrocnemius muscles of young and old rats, respectively. The data suggest that the denervation-induced muscle loss was at least partly due to apoptosis as indicated by elevated caspase 8 and Bax levels in denervated muscles. While Id2 may have a role in aging-induced sarcopenia, Id2 does not appear to directly regulate apoptosis during denervation. The elevated Id expression in muscles from aged animals is therefore not a direct consequence of loss of alpha-motoneurons during senescence. PMID- 12902527 TI - Effect of short-term and long-term antioxidant therapy on primary and secondary ageing neurovascular processes. AB - Previous studies from our laboratory demonstrated an age-related functional decline in sensory neurones and their modulation of microvascular blood flow (primary ageing processes) that correlated with a deficiency in tissue repair (a secondary ageing process). We also raised the notion of a possible role for free radicals in these age-related changes. The aim of this study was to investigate the impact of antioxidant therapy on modulating sensory neurovascular function and tissue repair with age. Twenty-four-month-old Sprague-Dawley rats were treated with vitamin E for short-term (40 mg/kg, i.p., every other day for 2 weeks) or long-term (for 12 months in advance, 10 g/kg, incorporated in food). These treated rats were assessed for the effectiveness of treatment and tested for their sensory neurovascular function, repair of full-thickness burn, and recovery from hyperalgesia following nerve injury. The results indicate that both short- and long-term vitamin E treatments are effective in improving sensory neurovascular function and in reducing the time required for complete wound closure of full-thickness burn injury. Short-term vitamin E treatment was more effective in protecting against the development of hyperalgesia following nerve injury. An initial increase in wound size and in hyperalgesia was observed in the treated animals, and could reflect possible side effects of the antioxidant therapy and support the importance of free radicals in early stages of the repair process. The data, overall, support the notion that oxidative damage contributes to both primary and secondary ageing processes. PMID- 12902528 TI - Mobility performance: a high-tech test for geriatricians. PMID- 12902529 TI - Walking performance and cardiovascular response: associations with age and morbidity--the Health, Aging and Body Composition Study. AB - BACKGROUND: The long-distance corridor walk is a timed 400-meter walk test used to assess walking endurance in well-functioning men and women aged 70-79 in the Health, Aging and Body Composition Study. METHODS: We examined walking time along with heart rate and blood pressure response in relationship to prevalent chronic conditions, weight, physical activity, and markers of subclinical disease. Of 3075 participants, 2324 (76%) completed the test with heart rate and blood pressure responses in the range expected for a moderate level of exertion. RESULTS: Slower walking time was influenced by both clinical and subclinical disease, and also was strongly related to both low and high body weight and low self-reported physical activity. Heart rate and blood pressure responses were higher with several clinical and subclinical diseases, but heart rate response and recovery were more strongly related to walking time than to disease. Higher body mass index and lower physical activity were associated with greater heart rate response and recovery. CONCLUSIONS: The independent contribution of both clinical and subclinical disease to walking time supports the use of walking tests as a summary measure of disease in older adults. The independent association of walking time with physical activity suggests that it is sensitive to levels of fitness as well. Together these findings show that walking performance is a valid indicator of physiologic reserve in older adults. PMID- 12902530 TI - Responsiveness of the index of mobility limitation: comparison with gait speed alone in the longitudinal aging study amsterdam. AB - BACKGROUND: Interpreting self-reported disability differences between diverse older populations is complicated by differences in attitudes and environment. We have previously reported on the index of mobility-related limitation tests (MOBLI), and shown that it predicts mortality over 4 years. In this article, we examine whether the index is responsive to changes in self-reported mobility disability. METHODS: Data on gait speed, time to complete 5 chair stands, and peak expiratory flow rate, with self-reported difficulty walking for 5 minutes, were available from the baseline and two 3-year follow-ups in the Longitudinal Aging Study Amsterdam. Analysis used data on changes in the index (or walking speed alone) and corresponding change over 3 years in self-reported difficulty or inability with a medium-distance walk. RESULTS: During all follow-ups, groups reporting deterioration in functioning had relatively larger changes in gait speed and MOBLI score than did the "no deterioration" groups. In comparative analyses of responsiveness, the MOBLI score had a larger responsiveness index, higher odds ratios, and larger receiving operating characteristic area than gait speed alone. CONCLUSIONS: The MOBLI index of mobility-related physical limitation tests is responsive to changes in self-reported mobility disability over two 3 year periods, and performs better than gait speed alone. This property is strongly supportive of its validity for epidemiological comparison of older populations across countries or over longer periods of time. PMID- 12902531 TI - A comparison of leg power and leg strength within the InCHIANTI study: which influences mobility more? AB - BACKGROUND: In a clinical study evaluating the functioning of mobility-limited elders, muscle power accounted for more of the variation in function than strength did. There was also evidence that the power-function relationship may be described as curvilinear. However, these findings have never been confirmed in a representative population. METHODS: An analysis was conducted using data from the InCHIANTI study, a population-based cohort of 1032 adults living in Italy. To assess the relationships between impairments in power and strength and mobility performance, we created separate multivariate linear and log-transformed regression models as well as separate logistic regression models. RESULTS: Subjects were age > or = 65 years, predominantly female (54%), with a mean age of 74.2 years, and most had mild-moderate mobility limitations (Short Physical Performance Battery score 10.5 +/- 2.1). Though leg extension power and isometric hip extension strength were closely associated, in several separate multivariate linear regression models, leg power consistently explained more of the variance than strength did in several measures of physical performance. Differences were even larger when observed in curvilinear models of power and strength. Using separate multivariate logistic regression models to examine the odds ratios for mobility limitations in persons with low muscle power versus high muscle power, and in persons with low muscle strength versus high muscle strength, we found that both factors influenced risk for mobility limitations, but low power was associated with a 2-3-fold greater likelihood than low strength. CONCLUSION: These findings identified muscle power to be a more influential proximal determinant of physical performance than impairments in strength and emphasized muscle power as an important determinant of mobility skills in older adults. PMID- 12902532 TI - Oxygen-uptake (VO2) kinetics and functional mobility performance in impaired older adults. AB - BACKGROUND: Measures of maximal oxygen uptake (VO(2max)) are limited in disabled older adults, and measures of submaximal oxygen uptake (VO(2)) may better predict functional mobility limitations. These measures may include oxygen-uptake kinetics at the onset of submaximal exercise or during recovery. We sought to determine whether the lag in oxygen uptake at the beginning of exercise (oxygen deficit) and excess oxygen uptake above rest following exercise (excess postexercise oxygen consumption) (a) predict physical performance in impaired older adults with decreased aerobic function, and (b) predict physical performance better than peak VO(2). METHODS: Two groups of community-dwelling volunteers aged 65 or older were recruited according to their performance on a maximal graded exercise test. Using the Social Security Administration criterion of disability of a peak VO(2) 18 (Unimpaired, n = 21, mean +/- SEM age 76 +/- 1 years). RESULTS: The mean +/- SEM peak VO(2) was 58% lower in the Impaired (14 +/- 1 ml/kg/min) than the Unimpaired (24 +/- 1 ml/kg/min) adults. The time constant for oxygen deficit, tc(deficit), was more than twice as high in the Impaired than the Unimpaired (p <.05), and the time constant for excess postexercise oxygen consumption, tc(EPOC), tended to be higher in the Impaired than the Unimpaired (by 43%, p =.09). Measures of submaximal oxygen-uptake kinetics were as strong or more strongly predictive of functional mobility performance than peak VO(2) in both Unimpaired and Impaired older adults. The major predictor of functional performance for the Unimpaired was a measure of oxygen deficit accruing during exercise (tc(deficit)), and for the Impaired, it was a measure of oxygen debt during recovery, tc(EPOC). CONCLUSIONS: Measurement of submaximal oxygen-uptake kinetics may provide a more practical and relevant assessment of deconditioning in frail older adults, and may eventually supplant maximal (peak) oxygen uptake as a predictor of functional disability in older adults. PMID- 12902533 TI - Effects of long-term resistive training on mobility and strength in older adults with diabetes. AB - BACKGROUND: Strength training has been shown to be beneficial in older adults. However, very little data exist on the effects of strength training in older diabetics. METHODS: 31 community-dwelling older adults with diabetes (mean age = 66.1 years) were randomly assigned to either an exercise (EX) or control (CO) group. The EX group trained the plantar flexors, knee extensors, knee flexors, hip extensors, and hip flexors muscle groups at 50%, 60%, and 70% of 1-repetition maximum, 2.6 days a week, for 24 months. Mobility tests included the timed up and go, 50-foot walk, and walking up and down 8 stairs. Strength and mobility for both groups were evaluated at 6-month intervals. RESULTS: There was a group and time effect as the EX group increased 31.4% (p <.001) in strength for all muscle groups after the first 6 months of training, and the strength gains were retained for the duration of the training intervention. There was also a group and time effect for mobility as performance increased 8.6% and 9.8% (p =.032 and p = 0.031) for the first 6 and 12 months, respectively, but decreased to 4.6% above baseline at the end of the intervention. There were essentially no changes from baseline strength or mobility values for the CO group. CONCLUSION: In conclusion, these data suggest that a moderate-intensity resistive-training program can improve mobility and strength for the duration of a 24-month intervention in older adults with diabetes, thus potentially reducing the rate of mobility loss during aging. PMID- 12902534 TI - Can neuromuscular strength and function in people with dementia be rehabilitated using resistance-exercise training? Results from a preliminary intervention study. AB - BACKGROUND: Neuromuscular weakness is a prominent symptom among people with central nervous system disorders, such as dementia, typically leading to disability in activities of daily life. We sought to evaluate the potential of resistance exercise to improve neuromuscular strength and function in the lower extremities among community-dwelling people with dementia. METHODS: Twenty-eight subjects, aged 70-88 years and with an average Mini-Mental State Examination score of 17.8 +/- 7.2, were recruited from a population of adult daycare facility users. Subjects underwent pre- and postintervention assessment of strength and physical function consisting of determination of bilateral maximum strength of the knee extensor, hip flexor, dorsiflexor muscles, and handgrip; and evaluation of lower-extremity function based on repeated chair stands and gait speed. The intervention consisted of moderate-intensity progressive resistance training of the hip extensors, abductors, knee extensors and flexors, and dorsiflexors using the Theraband resistance system for up to 3 days weekly over a 6-week period. RESULTS: Subjects completed an average of 11.4 +/- 2.5 exercise sessions. Among those who exercised at least twice per week (> or = 12), they improved an average of 15.6% in quadriceps strength, 10.1% in handgrip strength, 22.2% in sit-to stand (STS) time, 9.9% in usual gait time, 5.4% in fast gait time (p =.03), and 14.0% in the timed-up-and-go (TUG) test. CONCLUSIONS: Subjects demonstrated improvements in some areas of muscular capacity (quadriceps and handgrip) and most tests of lower-extremity function (STS, gait, TUG), yet declines in other areas (dorsiflexion and iliopsoas strength). Although strength or functional deficits in all domains were not remediable, these results suggest the potential of a resistance-exercise intervention of longer duration and/or greater intensity to produce beneficial effects on the neuromuscular functioning of people with dementia. PMID- 12902535 TI - Urinary incontinence: bridging the gender gap. PMID- 12902536 TI - Predictors of institutionalization in an older population during a 13-year period: the effect of urge incontinence. AB - BACKGROUND: Longitudinal data on predictors of institutionalization in random older populations are limited. The aim here was to identify predictors of institutionalization in an unselected older population during a period of 13 years with a special focus on the prognostic value of urge incontinence. METHODS: A population-based prospective survey was conducted involving 366 men and 409 women aged 60 years and older. Age-adjusted and multivariate Cox proportional hazards models were used to examine the predictive association of urge incontinence, living arrangements, neurological, cardiovascular, musculoskeletal, and other chronic diseases, activities of daily living (ADL) disability, and depressive symptoms with institutionalization separately in men and women. RESULTS: Adjusted for age, ADL disability and other chronic diseases predicted institutionalization in both men and women. Urge incontinence and depressive symptoms in men and living alone and cardiovascular diseases in women were also significant predictors. In multivariate analyses where all potential predictors were included simultaneously, age (RR [relative risk] 1.15; 95% CI [confidence interval] 1.10-1.19), urge incontinence (RR 3.07; 95% CI 1.24-7.59), and depressive symptoms (RR 1.22; 95% CI 1.00-1.48) remained significant predictors of institutionalization in men. In women, age (RR 1.15; 95% CI 1.12-1.19) and living alone (RR 2.02; 95% CI 1.27-3.21) were independent predictors. CONCLUSIONS: In addition to age, urge incontinence and depressive symptoms in men and living alone in women are significant prognostic indicators of institutionalization. The greater prognostic value of urge incontinence in men compared with women emphasizes the importance of interventions aimed at promoting continence and coping with the problem both at the individual and caregiver levels especially among older men. PMID- 12902537 TI - Taiji training improves knee extensor strength and force control in older adults. AB - The purpose of this study was to examine the effects of Taiji training on knee extensor strength and force control in older individuals. Twenty-six healthy older adults (71.9 +/- 1.8 years) participated in the study. Sixteen of the older adults (70.2 +/- 1.8 years) underwent Taiji training for 20 weeks (experimental group), whereas the other 10 (74.6 +/- 1.2 years) served as the control group. For both groups, strength and force control of the knee extensors was assessed twice with an isokinetic dynamometer. Strength was assessed with a maximum voluntary isometric contraction (MVC). Force control was measured as the standard deviation (SD) and coefficient of variation (CV) of force during a constant isometric knee extension task at 2%, 30%, 60%, and 90% MVC. For the experimental group, MVC significantly increased (19.5 +/- 4.9%) and the CV of force decreased (18.9 +/- 3.3%) following Taiji training. Improvements in the CV of force were primarily due to decreases in the SD of force (R(2) =.86) rather than increases in strength (R(2) =.12). Furthermore, decreases in SD of force were independent of improvements in strength. For the control group, strength, SD, and CV of force were not different for the 2 tests. The overall findings suggest that Taiji training improves knee extensor strength and force control in older adults. PMID- 12902538 TI - Five clinical tests to assess balance following ball exercises and treadmill training in adult persons with intellectual disability. AB - BACKGROUND: Incidence rates of falling increase progressively with aging. Preventing or delaying the onset of functional decline is a crucial important goal, because more individuals with intellectual disability (ID) are living well into their sixth and seventh decades. The question of whether walking and ball exercises can effect balance performance has never been reported. This pilot study was conducted to determine the effects of therapeutic training on improving balance capabilities in adults with mild ID. METHODS: The study included 13 women and 4 men, aged 50-67 years (mean age 56.5 years) residing in a residential care center. Five clinical tests were used to determine the "real" picture of the locomotor function and balance before and after the training protocol. Baseline values were determined using 2 control groups of age-matched adults with and without ID. The tests included modified get-up-and-go, full turn, forward reach, sit-to-stand, and one-legged standing. Therapeutic training for 6 months included dynamic ball exercises and treadmill walking with a 2-3% positive inclination. RESULTS: Participants in the program showed little to no improvement in terms of their static and dynamic balance compared to their initial values. Thus, only 2 of the tests showed statistical significance. CONCLUSIONS: Lack of improvement was noted in both postural and balance control in adults with mild ID as a result of 6 months of intervention by means of ball exercise and treadmill training. PMID- 12902540 TI - MicroRNAs and small interfering RNAs can inhibit mRNA expression by similar mechanisms. AB - MicroRNAs (miRNAs) are endogenously encoded small noncoding RNAs, derived by processing of short RNA hairpins, that can inhibit the translation of mRNAs bearing partially complementary target sequences. In contrast, small interfering RNAs (siRNAs), which are derived by processing of long double-stranded RNAs and are often of exogenous origin, degrade mRNAs bearing fully complementary sequences. Here, we demonstrate that an endogenously encoded human miRNA is able to cleave an mRNA bearing fully complementary target sites, whereas an exogenously supplied siRNA can inhibit the expression of an mRNA bearing partially complementary sequences without inducing detectable RNA cleavage. These data suggest that miRNAs and siRNAs can use similar mechanisms to repress mRNA expression and that the choice of mechanism may be largely or entirely determined by the degree of complementary of the RNA target. PMID- 12902539 TI - Proteasome inhibitors disrupt the unfolded protein response in myeloma cells. AB - Novel agents that target the proteasome, a proteolytic complex responsible for the degradation of ubiquitinated proteins, have demonstrated remarkable therapeutic efficacy in multiple myeloma, a plasma cell malignancy. However, the mechanism by which these compounds act remains unknown. A signaling pathway called the unfolded protein response (UPR) allows cells to handle the proper folding of proteins. The transcription factor XBP-1, a regulator of the UPR, is also required for plasma cell differentiation, suggesting a link between the UPR and plasma cell differentiation. Here we show that proteasome inhibitors target XBP-1 and the UPR in myeloma cells. Proteasome inhibitors suppress the activity of the translumenal endoplasmic reticulum endoribonuclease/kinase, IRE1 alpha, to impair the generation of the active, spliced XBP-1 species and simultaneously stabilize the unspliced species that acts as a dominant negative. Myeloma cells rendered functionally deficient in XBP-1 undergo increased apoptosis in response to endoplasmic reticulum stress. Identification of compounds that target the activity of IRE1 alpha/XBP-1 may yield novel therapies for the treatment of multiple myeloma and other malignancies that rely on an intact UPR. PMID- 12902541 TI - Single-molecule study of DNA unlinking by eukaryotic and prokaryotic type-II topoisomerases. AB - Type-II topoisomerases are responsible for untangling DNA during replication by removing supercoiled and interlinked DNA structures. Using a single-molecule micromanipulation setup, we follow the real-time decatenation of two mechanically braided DNA molecules by Drosophila melanogaster topoisomerase (Topo) II and Escherichia coli Topo IV. Although Topo II relaxes left-handed (L) and right handed (R-) braids similarly at a rate of approximately 2.9 s-1, Topo IV has a marked preference for L-braids, which it relaxes completely and processively at a rate of approximately 2.4 s-1. However, Topo IV can unlink R-braids at about half that rate when they supercoil to form L-plectonemes. These results imply that the preferred substrate for unlinking by Topo IV has the symmetry of an L-crossing and shed new light on the decatenation of daughter strands during DNA replication, which are usually assumed to be linked in an R-braid. PMID- 12902543 TI - Shrinkage-based similarity metric for cluster analysis of microarray data. AB - The current standard correlation coefficient used in the analysis of microarray data was introduced by M. B. Eisen, P. T. Spellman, P. O. Brown, and D. Botstein [(1998) Proc. Natl. Acad. Sci. USA 95, 14863-14868]. Its formulation is rather arbitrary. We give a mathematically rigorous correlation coefficient of two data vectors based on James-Stein shrinkage estimators. We use the assumptions described by Eisen et al., also using the fact that the data can be treated as transformed into normal distributions. While Eisen et al. use zero as an estimator for the expression vector mean mu, we start with the assumption that for each gene, mu is itself a zero-mean normal random variable [with a priori distribution N(0,tau 2)], and use Bayesian analysis to obtain a posteriori distribution of mu in terms of the data. The shrunk estimator for mu differs from the mean of the data vectors and ultimately leads to a statistically robust estimator for correlation coefficients. To evaluate the effectiveness of shrinkage, we conducted in silico experiments and also compared similarity metrics on a biological example by using the data set from Eisen et al. For the latter, we classified genes involved in the regulation of yeast cell-cycle functions by computing clusters based on various definitions of correlation coefficients and contrasting them against clusters based on the activators known in the literature. The estimated false positives and false negatives from this study indicate that using the shrinkage metric improves the accuracy of the analysis. PMID- 12902542 TI - Horizontal gene transfer: a critical view. AB - It has been suggested that horizontal gene transfer (HGT) is the "essence of phylogeny." In contrast, much data suggest that this is an exaggeration resulting in part from a reliance on inadequate methods to identify HGT events. In addition, the assumption that HGT is a ubiquitous influence throughout evolution is questionable. Instead, rampant global HGT is likely to have been relevant only to primitive genomes. In modern organisms we suggest that both the range and frequencies of HGT are constrained most often by selective barriers. As a consequence those HGT events that do occur most often have little influence on genome phylogeny. Although HGT does occur with important evolutionary consequences, classical Darwinian lineages seem to be the dominant mode of evolution for modern organisms. PMID- 12902544 TI - Nonparametric regression to the mean. AB - Available data may reflect a true but unknown random variable of interest plus an additive error, which is a nuisance. The problem in predicting the unknown random variable arises in many applied situations where measurements are contaminated with errors; it is known as the regression-to-the-mean problem. There exists a well known solution when both the distributions of the true underlying random variable and the contaminating errors are normal. This solution is given by the classical regression-to-the-mean formula, which has a data-shrinkage interpretation. We discuss the extension of this solution to cases where one or both of these distributions are unknown and demonstrate that the fully nonparametric case can be solved for the case of small contaminating errors. The resulting nonparametric regression-to-the-mean paradigm can be implemented by a straightforward data-sharpening algorithm that is based on local sample means. Asymptotic justifications and practical illustrations are provided. PMID- 12902547 TI - Elucidating the molecular mechanism of cardiac remodeling using a comparative genomic approach. AB - It is proposed that analysis of global gene expression would provide an understanding of the molecular mechanisms of cardiac remodeling. However, previous studies have only provided "snapshots" of differential gene expression. Furthermore, the differences in gene expression between regions of the heart that can result in sampling variability have not been characterized. In this study, we employed the Affymetrix GeneChip technology to evaluate the patterns of expression in two different in vivo models of cardiac remodeling and in two different regions (left ventricle free wall and intraventricular septum) of the heart. Mice underwent transverse aortic constriction (TAC), myocardial infarction (MI), or sham operation, and RNA from the left ventricle free wall and the septum was isolated 1 wk later. Histological analysis showed profound myocyte hypertrophy and fibrosis in both the septum and the left ventricle free wall of the TAC model, whereas, in the MI model, only the left ventricle exhibited hypertrophy. These differences were also reflected in the expression analysis. In conclusion, our analysis shows that regional differences in gene expression exist in the heart. Moreover, common pathways that are coregulated in both models exist, and these might be central to the hypertrophic phenotype regardless of the initial hypertrophic stimuli. PMID- 12902546 TI - Gene expression profiling and functional proteomic analysis reveal perturbed kinase-mediated signaling in genetic stroke susceptibility. AB - The stroke-prone spontaneously hypertensive rat (SHRSP) is a model of heritable hypertension-associated cerebrovascular injury. This study sought to compare SHRSP to the stroke-resistant SHR strain to identify genes and protein pathways whose expression and/or function was significantly altered between the strains prior to the onset of stroke. Cerebral cortex gene expression profiles from male SHRSPs and matched SHRs were examined by Affymetrix microarray analysis. mRNAs encoding the brain-derived neurotrophic factor receptor (TrkB) and multiple kinases of the MAPK/AKT signaling pathways, including JNK2, AKT2, and PI3K, were differentially expressed between SHRSP and SHR. Because these data suggest altered function in pathways involving MAP and AKT kinase activity, we performed Western blot using phosphorylation state-specific antibodies to characterize activity of MAP kinase and PI3K/AKT pathways. Changes in the levels of the phosphorylated forms of these kinases paralleled the changes in transcript levels observed between the strains. Two-dimensional gel electrophoresis and peptide fragment mass fingerprinting were used to identify altered protein substrates of these kinases. Protein profiling of kinase substrates further supported the notion of perturbed kinase-mediated signaling in SHRSP and identified adenylyl cyclase associated protein 2, TOAD-64, propionyl CoA carboxylase, APG-1, and valosin-containing protein as kinase targets whose phosphorylation state is altered between these strains. Altered gene and protein expression patterns in SHRSP are consistent with increased vulnerability of this strain to cerebrovascular injury. PMID- 12902545 TI - The origin and liver repopulating capacity of murine oval cells. AB - The appearance of bipotential oval cells in chronic liver injury suggests the existence of hepatocyte progenitor/stem cells. To study the origin and properties of this cell population, oval cell proliferation was induced in adult mouse liver by 3,5-diethoxycarbonyl-1,4-dihydrocollidine (DDC) and a method for their isolation was developed. Transplantation into fumarylacetoacetate hydrolase (Fah) deficient mice was used to determine their capacity for liver repopulation. In competitive repopulation experiments, hepatic oval cells were at least as efficient as mature hepatocytes in repopulating the liver. In mice with chimeric livers, the oval cells were not derived from hepatocytes but from liver nonparenchymal cells. This finding supports a model in which intrahepatic progenitors differentiate into hepatocytes irreversibly. To determine whether oval cells originated from stem cells residing in the bone marrow, bone marrow transplanted wild-type mice were treated with DDC for 8 months and oval cells were then serially transferred into Fah mutants. The liver repopulating cells in these secondary transplant recipients lacked the genetic markers of the original bone marrow donor. We conclude that hepatic oval cells do not originate in bone marrow but in the liver itself, and that they have valuable properties for therapeutic liver repopulation. PMID- 12902548 TI - Molecular mechanisms of reduced beta-adrenergic signaling in the aged heart as revealed by genomic profiling. AB - Myocardial aging leads to a reduction of beta-adrenergic receptor-induced metabolic and contractile responsiveness. We hypothesize that a change in the patterns of gene expression is important in these age-related events. To test this, hearts were harvested from young and aged male rats (3-4 and 20-22 mo, respectively). Total mRNA was extracted and prepared for hybridization to Affymetrix U34A GeneChips. Filtering criteria, involving fold change and a statistical significance cutoff were employed, yielding 263 probe pairs exhibiting differential signals. Of the 163 annotated genes, at least 56 (34%) were classified as signaling/cell communication. Of these 56, approximately half were directly involved in G protein-coupled receptor signaling pathways. We next determined which of these changes might be involved in anti-adrenergic activity and identified 19 potentially important gene products. Importantly, we observed a decrease in beta1-adrenergic receptor and adenylyl cyclase mRNAs, whereas the mRNA encoding beta-arrestin increased. Furthermore, the results demonstrate an increase in mRNAs encoding the adenosine A1 receptor and phospholipase D, which could increase anti-adrenergic effects. Moreover, the mRNAs encoding the muscarinic M3 receptor, nicotinic acetylcholine receptor beta3, and nicotinic acetylcholine receptor-related protein were increased as was the mRNA encoding guanylate kinase-associated protein. Interestingly, we also observed eight mRNAs whose abundance changed three- to sixfold with aging that could be considered as being compensatory. Although these results do not prove causality, they demonstrate that cardiac aging is associated with changes in the profiles of gene expression and that many of these changes may contribute to reduced adrenergic signaling. PMID- 12902549 TI - Identification of new genes differentially expressed in coronary artery disease by expression profiling. AB - Genetic factors increase the risk to coronary artery disease (CAD). To date, a limited number of genes that potentially contribute to development of CAD have been identified. In this study, we have performed large-scale gene expression analysis of approximately 12,000 human genes in nine severely atherosclerotic and six nonatherosclerotic human coronary arteries using oligonucleotide microarrays. Fifty-six genes showed differential expression in atherosclerotic coronary artery tissues; expression of 55 genes was increased in atherosclerotic coronary arteries, whereas only one gene, GST, encoding a reducing agent, showed downregulated expression. The expression data of selected genes were validated by quantitative RT-PCR analysis as well as immunostaining. The associations of 49 genes with CAD appear to be novel, and they include genes encoding ICAM-2, PIM-2, ECGF1, fusin, B cell activator (BL34, GOS8), Rho GTPase activating protein-4, retinoic acid receptor responder, beta2-arrestin, membrane aminopeptidase, cathepsins K and H, MIR-7, TNF-alpha-induced protein 2 (B94), and flavocytochrome 558. In conclusion, we have identified 56 genes whose expression is associated with CAD, and 49 of them may represent new genes linked to CAD. PMID- 12902551 TI - Transit peptide cleavage sites of integral thylakoid membrane proteins. AB - A set of 58 nuclearly encoded thylakoid-integral membrane proteins from four plant species was identified, and their amino termini were assigned unequivocally based upon mass spectrometry of intact proteins and peptide fragments. The dataset was used to challenge the Web tools ChloroP, TargetP, SignalP, PSORT, Predotar, and MitoProt II for predicting organelle targeting and transit peptide proteolysis sites. ChloroP and TargetP reliably predicted chloroplast targeting but only reliably predicted transit peptide cleavage sites for soluble proteins targeted to the stroma. SignalP (eukaryote settings) accurately predicted the transit peptide cleavage site for soluble proteins targeted to the lumen. SignalP (Gram-negative bacteria settings) reliably predicted peptide cleavage of integral thylakoid proteins inserted into the membrane via the "spontaneous" pathway. The processing sites of more common thylakoid-integral proteins inserted by the signal recognition peptide-dependent pathway were not well predicted by any of the programs. The results suggest the presence of a second thylakoid processing protease that recognizes the transit peptide of integral proteins inserted via the spontaneous mechanism and that this mechanism may be related to the secretory mechanism of Gram-negative bacteria. PMID- 12902552 TI - The radiation dose in interventional radiology study: knowledge brings responsibility. PMID- 12902553 TI - Percutaneous vertebroplasty: technical considerations. AB - Percutaneous vertebroplasty has now been in use in the United States for 10 years. Standards of practice are available from the American College of Radiology. The performance of this procedure is becoming routine for most interventional practices, and it has become the general standard of care for pain associated with vertebral compression fractures of the spine. This article describes routine procedural technique and considerations and is intended to review the authors' knowledge of the procedure and provide reliable methods for good outcomes and complication avoidance. PMID- 12902554 TI - A practical approach to hypertension in the 21st century. PMID- 12902555 TI - Radiation doses in interventional radiology procedures: the RAD-IR study: part II: skin dose. AB - PURPOSE: To determine peak skin dose (PSD), a measure of the likelihood of radiation-induced skin effects, for a variety of common interventional radiology and interventional neuroradiology procedures, and to identify procedures associated with a PSD greater than 2 Gy. MATERIALS AND METHODS: An observational study was conducted at seven academic medical centers in the United States. Sites prospectively contributed demographic and radiation dose data for subjects undergoing 21 specific procedures in a fluoroscopic suite equipped with built-in dosimetry capability. Comprehensive physics evaluations and periodic consistency checks were performed on each unit to verify the stability and consistency of the dosimeter. Seven of 12 fluoroscopic suites in the study were equipped with skin dose mapping software. RESULTS: Over a 3-year period, skin dose data were recorded for 800 instances of 21 interventional radiology procedures. Wide variation in PSD was observed for different instances of the same procedure. Some instances of each procedure we studied resulted in a PSD greater than 2 Gy, except for nephrostomy, pulmonary angiography, and inferior vena cava filter placement. Some instances of transjugular intrahepatic portosystemic shunt (TIPS) creation, renal/visceral angioplasty, and angiographic diagnosis and therapy of gastrointestinal hemorrhage produced PSDs greater than 3 Gy. Some instances of hepatic chemoembolization, other tumor embolization, and neuroembolization procedures in the head and spine produced PSDs greater than 5 Gy. In a subset of 709 instances of higher-dose procedures, there was good overall correlation between PSD and cumulative dose (r = 0.86; P <.000001) and between PSD and dose area-product (r = 0.85, P <.000001), but there was wide variation in these relationships for individual instances. CONCLUSIONS: There are substantial variations in PSD among instances of the same procedure and among different procedure types. Most of the procedures observed may produce a PSD sufficient to cause deterministic effects in skin. It is suggested that dose data be recorded routinely for TIPS creation, angioplasty in the abdomen or pelvis, all embolization procedures, and especially for head and spine embolization procedures. Measurement or estimation of PSD is the best method for determining the likelihood of radiation-induced skin effects. Skin dose mapping is preferable to a single-point measurement of PSD. PMID- 12902556 TI - Endovenous laser treatment of saphenous vein reflux: long-term results. AB - PURPOSE: To report long-term follow-up results of endovenous laser treatment for great saphenous vein (GSV) reflux caused by saphenofemoral junction (SFJ) incompetence. MATERIALS AND METHODS: Four hundred ninety-nine GSVs in 423 subjects with varicose veins were treated over a 3-year period with 810-nm diode laser energy delivered percutaneously into the GSV via a 600- micro m fiber. Tumescent anesthesia (100-200 mL of 0.2% lidocaine) was delivered perivenously under ultrasound (US) guidance. Patients were evaluated clinically and with duplex US at 1 week, 1 month, 3 months, 6 months, 1 year, and yearly thereafter to assess treatment efficacy and adverse reactions. Compression sclerotherapy was performed in nearly all patients at follow-up for treatment of associated tributary varicose veins and secondary telangiectasia. RESULTS: Successful occlusion of the GSV, defined as absence of flow on color Doppler imaging, was noted in 490 of 499 GSVs (98.2%) after initial treatment. One hundred thirteen of 121 limbs (93.4%) followed for 2 years have remained closed, with the treated portions of the GSVs not visible on duplex imaging. Of note, all recurrences have occurred before 9 months, with the majority noted before 3 months. Bruising was noted in 24% of patients and tightness along the course of the treated vein was present in 90% of limbs. There have been no skin burns, paresthesias, or cases of deep vein thrombosis. CONCLUSIONS: Long-term results available in 499 limbs treated with endovenous laser demonstrate a recurrence rate of less than 7% at 2 year follow-up. These results are comparable or superior to those reported for the other options available for treatment of GSV reflux, including surgery, US guided sclerotherapy, and radiofrequency ablation. Endovenous laser appears to offer these benefits with lower rates of complication and avoidance of general anesthesia. PMID- 12902557 TI - Subintimal angioplasty of long superficial femoral artery occlusions. AB - PURPOSE: To investigate the value of subintimal angioplasty (SA) and selective stent placement in the treatment of long (> or =15 cm) superficial femoral artery (SFA) occlusions. MATERIALS AND METHODS: During a period of 4.5 years, 67 long SFA occlusions in 61 patients (52 male, 9 female) were intended to be treated with SA, either retrogradely (n = 55) or antegradely (n = 12). Postprocedural medical treatment included aspirin + ticlopidine/clopidogrel (AT/C) combination. In 25 patients warfarin was also given for 3-6 months. Patients were followed up for 1-30 months (mean 12.5 +/- 9.0 months). Hemodynamic patencies were determined with the Kaplan-Meier method, risk factors affecting patency were evaluated with the Cox model, and the patencies of the subgroups were compared with log-rank test. RESULTS: Subintimal recanalization was technically successful in 59 of 67 occlusions. Technical success was 83% in the first 30 procedures, 92% in the last 37, and 100% in the last 29. Forty-six occlusions were treated with SA alone and 13 with SA and stent placement. On an intention-to-treat basis, primary patency at 6 and 12 months was 49% and 22%, respectively, and assisted primary patency at 6 and 12 months was 69% and 57%, respectively. Patency rates were not significantly different in patients with claudication versus critical limb ischemia, or in those treated with SA alone versus SA and stent placement. With the multivariate Cox model, medical treatment with AT/C combination was identified as the only significant risk factor for both primary patency and assisted primary patency. With the Kaplan-Meier analysis, primary and assisted primary patencies were significantly higher in the warfarin group than the AT/C group (P =.0002 and.0001, respectively). CONCLUSION: SA is a simple and safe method with a high technical success rate in the endovascular treatment of long SFA occlusions. Long-term patency rates, however, seem unsatisfactory, despite early reports. Subintimal stent placement provides cumulative patency at least as good as SA alone. Warfarin may significantly improve both primary patency and assisted primary patency after subintimal recanalization, but even with this treatment patency rates are still lower than those reported for bypass surgery. Therefore, in long SFA occlusions, SA is not recommended for claudicants but may be valuable in patients with critical limb ischemia. PMID- 12902558 TI - Endovascular aneurysm repair with the Talent stent-graft. AB - PURPOSE: Evaluation of the Talent endovascular aortic stent graft in the management of abdominal aortic aneurysms. MATERIALS AND METHODS: Thirty-eight patients with suitable abdominal aortic aneurysms were treated over an 18-month period using the modular Talent stent graft. The suprarenal Talent device was deployed in 31 cases. Clinical follow-up of all patients has been performed by clinical examination, contrast-enhanced CT, and Duplex ultrasound for a mean period of 12.5 months. RESULTS: Graft placement was successful in all 38 patients. The immediate exclusion rate was 84%, the 1-month primary exclusion rate was 92.1%, and the 3-month exclusion rate was 97%. There were no deaths in the first 30 days, one death at 3 months due to a presumed rupture, and one other death at 1 year due to carcinomatosis. There have been no migrations or renal complications in the suprarenal group. CONCLUSIONS: Our short- and medium-term results are comparable with other published Talent stent-graft series. Suprarenal graft fixation is a safe procedure that may aid in preventing graft migration. Long-term follow-up is required to assess the durability of the suprarenal Talent device. PMID- 12902559 TI - Gadolinium, carbon dioxide, and iodinated contrast material for planning inferior vena cava filter placement: a prospective trial. AB - PURPOSE: To prospectively compare the diagnostic accuracy of CO(2) and gadolinium to iodinated contrast material for inferior vena cavography before inferior vena cava (IVC) filter placement. MATERIALS AND METHODS: Forty patients underwent injection of iodinated contrast material, CO(2), and gadolinium. Iodinated contrast material was used as the standard. Caval diameter was determined with calibrated software. Three readers blinded to contrast agent used measured the distance from the superior image border to the inferior margin of the renal veins and from the inferior image border to the iliac bifurcation. The measurements with CO(2) and gadolinium were compared to those with iodinated contrast material to obtain the interobserver and intraobserver variability. The presence or absence of caval thrombus and variant anatomy was noted. The same readers reexamined 12 studies in a separate session to determine intraobserver variability and correlation. RESULTS: Caval diameter differed by 0.4 mm or less for all three agents. Measurements with all agents were within 2 mm of each other for all patients. Gadolinium and CO(2) were not significantly different from one another in measuring caval diameter. At the initial reading, compared with iodinated contrast material, gadolinium had greater mean interobserver error in measuring the distance to the iliac bifurcation and both renal veins (range, 1.6 1.8 mm) than CO(2) (range, 0.2-1.4 mm). This finding, although statistically significant for gadolinium (P <.05), was of doubtful clinical relevance. Interobserver correlation was significantly worse for CO(2) at the levels of the iliac bifurcation (P =.02) and right renal vein (P =.008). Interobserver correlation for gadolinium was similar to that for iodinated contrast material at all levels. At repeat reading, there was significantly inferior intraobserver correlation with use of CO(2) for both renal veins (P <.05) compared to iodinated contrast material and for the left renal vein (P <.05) compared to gadolinium. Gadolinium identified three of three renal vein anomalies identified with iodinated contrast material whereas CO(2) localized one of three. CONCLUSION: CO(2) and gadolinium had limitations when compared with iodinated contrast material. Gadolinium provided superior consistency in identifying relevant landmarks for filter placement. CO(2) demonstrated significantly greater mean correlative error than gadolinium at initial and repeat readings. PMID- 12902560 TI - Semitransparent volume-rendering CT angiography for lesion display in aortoiliac arteriosclerotic disease. AB - PURPOSE: The potential of semitransparent volume-rendering (STVR) computed tomographic (CT) angiography was evaluated for the assessment of hemodynamically significant stenoses in aortoiliac arteriosclerotic disease. MATERIALS AND METHODS: In a prospective study, 76 patients (57 men, 19 women; mean age, 70 years) underwent single-detector (n = 26) or multidetector (n = 50) CT angiography of the aortoiliac region. Intraarterial digital subtraction angiography (DSA) was performed in each patient. STVR images with semitransparent display of arterial lumen (opacity, 50%) and vascular calcifications (opacity, 20%), as well as maximum-intensity projection (MIP), frontal/sagittal curved planar reformation (CPR), and MIP/axial studies were independently reviewed for hemodynamically significant lesions (> or =70% cross-sectional area reduction). DSA combined with invasive pressure measurement was used as the standard of reference. Vessel wall calcifications were classified according to a four-point scale (0, not calcified; 1, mildly calcified; 2, moderately calcified; 3, severely calcified). RESULTS: Of the 380 reviewed vessel sections, 28 represented 70%-99% stenoses and 14 represented occlusions. For detecting >/==" BORDER="0">70% lesions, STVR as well as CPR and MIP/axial studies revealed significantly higher specificity (91%-94%), positive predictive value (PPV; 0.62 0.72), and accuracy (90%-94%) than MIP (specificity, 59%; PPV, 0.27; accuracy, 64%; P <.001) in moderately or severely calcified vessel sections. In noncalcified or mildly calcified sections, the specificity of MIP was comparable with the other rendering techniques (96%-98%; P values ranging from.34 to.77). No significant sensitivity differences were noticed among the CT angiography modalities: STVR, 79%; MIP, 88%; CPR, 83%; and MIP/axial, 93% (P values ranging from.12 to.78). Negative predictive values ranged from 0.97 to 0.99 (P values ranging from.14 to 1). Median calcification scores in sections with overestimated lumen narrowings were significantly higher (3, severely calcified) than in sections with correctly graded lumen width (1, mildly calcified) with all CT angiography modalities (P <.05). CONCLUSION: With use of STVR, visualization of vascular lumen can be improved by rendering highly transparent mural calcifications. Hence, for three-dimensional presentation of aortoiliac arteriosclerotic disease, STVR studies should be preferred to MIPs as supplements to review of axial-source images. PMID- 12902561 TI - National trends in gastrointestinal access procedures: an analysis of Medicare services provided by radiologists and other specialists. AB - PURPOSE: To evaluate national trends in enteral access services by radiologists and other specialists. MATERIALS AND METHODS: Medicare data from 1997 to 2000 were analyzed for trends in gastrointestinal access services. Current Procedural Terminology codes for gastrostomy placement and maintenance services were selected. Utilization was analyzed by physician specialty. Targeted service analysis was performed for interventional radiologists. RESULTS: For sampled enteral access procedures, annual services to Medicare beneficiaries increased from 279,509 to 283,353 (+1.4%). These were most often performed by gastroenterologists (48.6%), surgeons (25.1%), radiologists (7.4%), and others (18.9%). Total procedures by radiologists increased 29.6% whereas procedures by gastroenterologists, surgeons, and other nonradiologists changed +6.9%, -4.9%, and -10.2%, respectively. For new gastrostomy accesses, radiologist volume increased 46.9% whereas gastroenterologist, surgeon, and other volumes changed +7.9%, -5.0%, and -21.5%, respectively. For maintenance services, radiologist volume increased 21.8% whereas gastroenterologist, surgeon, and other volumes changed +3.1%, -4.7%, and +7.9%, respectively. Analyzed for frequency, relative value, and physician time, enteral access services account for less than 1% of all services provided by interventional radiologists. CONCLUSIONS: Although the number of gastrointestinal access services provided to Medicare beneficiaries has remained static, radiologists have experienced a marked relative increase in volume, particularly for new gastrostomy procedures. This increase is largely at the expense of surgeons and other nongastroenterologists. However, radiologists still provide only a small portion of gastrointestinal access services nationwide, and these services account for only a small portion of all procedures performed by interventionalists. Therefore, the potential for enteral access service growth in interventional radiology is high. PMID- 12902562 TI - Percutaneous radiofrequency ablation of lung tumors in a large animal model. AB - PURPOSE: Percutaneous radiofrequency ablation (RFA) is accepted therapy for liver tumors in the appropriate clinical setting, but its use in lung neoplasms remains investigational. We undertook this study to evaluate the feasibility and immediate effectiveness of RFA for treatment of both solitary pulmonary nodules and clusters of lung tumors in a large animal model. MATERIALS AND METHODS: Percutaneous RFA of 14 lung tumors in five dogs was performed under CT guidance. Animals were euthanatized 8-48 hours after the procedure. The lungs and adjacent structures were harvested for gross and histopathologic evaluation. RESULTS: Five solitary pulmonary nodules (range, 17-26 mm) and three clusters of three nodules each (range, 7-17 mm per nodule) were treated with RFA. All ablations were technically successful. Perilesional ground-glass opacity and small asymptomatic pneumothoraces (n = 4) were visualized during the RFA sessions. One dog developed a large pneumothorax treated with tube thoracostomy but was euthanatized 8 hours post-RFA for persistent pneumothorax and continued breathing difficulty. Follow up CT 48 hours post-RFA revealed opacification of the whole lung segment. Gross and histopathologic evaluation showed complete thermal coagulation necrosis of all treated lesions without evidence of any viable tumor. The region of thermal coagulation necrosis typically extended to the lung surface. Small regions of pulmonary hemorrhage and congestion often surrounded the areas of coagulation necrosis. CONCLUSIONS: RFA can be used to treat both solitary pulmonary nodules and clusters of tumor nodules in the canine lung tumor model. This model may be useful for development of specific RFA protocols for human lung tumors. PMID- 12902563 TI - Image-guided percutaneous chemical and radiofrequency tumor ablation in an animal model. AB - PURPOSE: To determine whether combining acetic acid instillation before radiofrequency (RF) ablation can improve local tissue electrical conductivity, RF energy deposition, intratumoral heating, and tumor necrosis in a large animal model. MATERIALS AND METHODS: Multiple hypovascular canine venereal sarcomas were implanted in 11 mildly immunosuppressed dogs (25 mg/kg cyclosporin A twice daily). Tumors were incubated for 8-12 weeks to 4.2 cm +/- 0.6 in diameter. Treatment strategies included 10% and 15% acetic acid diluted in distilled water, 10% and 15% acetic acid diluted in saturated NaCl solution, 50% acetic acid, and 100% ethanol, with 6 mL of each injected alone or in combination with RF ablation (internally cooled, 1-cm tip; 12 minutes). Two additional control groups were studied in which tumors received either RF alone or distilled water injected alone. Comparisons were also made with groups treated with 36% NaCl with and without RF ablation. Resultant coagulation for these ablative strategies, along with local temperatures and RF parameters such as impedance, current, and power, were compared. RESULTS: Increasing coagulation was observed with increasing acetic acid concentrations (1.7 cm +/- 0.4, 2.8 cm +/- 0.6, and 3.5 cm +/- 0.3 for 10%, 15%, and 50% acetic acid alone, respectively; P <.01). The combination of RF ablation with acetic acid resulted in greater coagulation than with either therapy alone (P <.05). However, maximum heating and coagulation were observed with 10% acetic acid diluted in NaCl, with which the entire tumor (diameter, 4.5 cm +/- 0.4) was completely ablated in every case. This was equivalent to results for tumors treated with 36% NaCl combined with RF. RF with a 50% acetic acid concentration resulted in coagulation measuring only 3.7 cm +/- 0.3 (P <.01). Significantly greater RF heating (89.7 degrees C +/- 12.3 at 10 mm) was observed when the tumors were pretreated with 10% or 15% acetic acid in saturated NaCl, compared with 67.9 degrees C +/- 13.7 observed when acetic acid was diluted in water (P <.02). RF combined with ethanol produced less coagulation (2.8 cm +/- 0.3) than combinations with acetic acid because rapid and irreversible impedance increases were observed. CONCLUSION: Addition of acetic acid injections to RF ablation substantially increases tumor destruction compared with RF or injection therapy alone. However, lower acetic acid concentrations in saturated NaCl produced greater tumor coagulation, suggesting that, in this hypovascular tumor model, alterations in electrical conductivity play a more important role in increasing tumor ablation efficiency than do the additional ablative effects of acetic acid. PMID- 12902565 TI - Placement of subcutaneous chest wall ports ipsilateral to axillary lymph node dissection. AB - Without scientific evidence, practice has been to avoid placing chest wall ports ipsilateral to axillary lymph node dissection. Because the lymphatic system in the chest wall drains via both the internal mammary and axillary nodes, it seems that this practice might unduly restrict venous access options. Our study was designed to evaluate outcome after placement of chest wall ports ipsilateral to axillary lymph node dissection in patients with breast cancer. Twenty-eight patients were studied in this retrospective two-institution review. The incidence of lymphedema after port placement (3.6%) was no higher than that reported after axillary lymph node dissection alone. PMID- 12902564 TI - Carbon-coated stents implanted in porcine iliac and renal arteries: histologic and histomorphometric study. AB - PURPOSE: To test in a pig model the biocompatibility and effectiveness of carbon coated renal and iliac artery stent systems during implantation procedures and at different follow-up periods. MATERIALS AND METHODS: Twenty-two miniature pigs received carbon-coated balloon-expandable stainless-steel stents in their renal and iliac arteries. Animals were killed at 7, 30, or 180 days for evaluation of acute, subacute, and chronic biologic response to the implanted devices. Histologic, histomorphometric, and scanning electron microscopy (SEM) analyses were performed to assess inflammatory reaction, endothelialization process, and neointimal growth. RESULTS: Forty-four iliac stents and 42 renal stents were successfully implanted. Mural thrombi were not observed by angiography or histologic examinations. Histologically, no significant inflammatory reaction was detected: the stents appeared covered by a thin monolayer of endothelial cells even at 7-day follow-up. The neointima showed homogeneous growth and moderate thickness at 30-day and 180-day explantations (0.09 mm +/- 0.06 and 0.15 mm +/- 0.13, respectively, for renal arteries; 0.12 mm +/- 0.04 and 0.21 mm +/- 0.12, respectively, for iliac arteries). Internal and external elastic laminae were intact in 82 of 86 cases (95%) of stent-implanted arteries. Histologic validations were obtained with SEM observations for each follow-up group. CONCLUSIONS: This study showed good technical results of deployment of carbon coated renal and iliac stents and very satisfactory biologic behavior in terms of tissue and hematologic compatibility. The devices do not induce thrombus formation. PMID- 12902566 TI - Delayed femoral vein thrombosis after ultrasound-guided thrombin injection of a postcatheterization pseudoaneurysm. AB - Ultrasound-guided thrombin injection is a highly effective therapy for postcatheterization pseudoaneurysm. Despite a very low complication rate, a number of severe arterial thrombotic events have been reported. We present an unusual case of acute femoral vein thrombosis occurring several hours after successful treatment of femoral artery pseudoaneurysm by ultrasound-guided thrombin injection. Pathophysiologic mechanisms are discussed. This case highlights the potential hazards of instilling such a powerful thrombogenic substance in the immediate vicinity of other vascular structures. PMID- 12902567 TI - Trial design and reporting standards for intraarterial cerebral thrombolysis for acute ischemic stroke. 2003. PMID- 12902568 TI - G beta gamma and KACh: old story, new insights. AB - Dissociation of the heterotrimeric GTP-binding protein (G protein) betagamma subunits from the alpha subunit is a prerequisite step in the ability of these proteins to signal to downstream effectors. There is evidence that ions such as Na+ and Cl- can facilitate this dissociation. Interestingly, for KACh, the first known effector for Gbetagamma, intracellular Na+ can also activate the channel independently of Gbetagamma. Both Gbetagamma and Na+ strengthen channel interactions with the membrane phospholipid phosphatidylinositol 4,5-bisphosphate (PIP2), an event thought to be essential in opening the channel. PIP2 interacts with channel regions that form a binding pocket proximal to the transmembrane domains and is likely to exert a tangential, pulling force to mechanically open a gate at the cytoplasmic face of the channel pore. The tangential force generated by channel-PIP2 interactions is the likely force behind gating in all inwardly rectifying K+ channels. The gate opens when the lower part of the pore-lining transmembrane alpha helix pivots around a glycine residue in the middle of the helix. This mechanism of channel gating is conserved among K+ channels from bacteria to mammals and may represent a common mechanism for K+ channel gating. PMID- 12902570 TI - Don't overlook condoms for HIV prevention. PMID- 12902571 TI - Randomised controlled trial and economic evaluation of podophyllotoxin solution, podophyllotoxin cream, and podophyllin in the treatment of genital warts. AB - OBJECTIVES: To evaluate the efficacy and cost effectiveness of self applied podophyllotoxin 0.5% solution and podophyllotoxin 0.15% cream, compared to clinic applied 25% podophyllin in the treatment of genital warts over 4 weeks. METHODS: We conducted a randomised controlled trial in 358 immunocompetent men and women with genital warts of 3 months' duration or less. RESULTS: In the principal analysis both podophyllotoxin solution (OR 2.93, 95% CI 1.56 to 5.50) and podophyllotoxin cream (OR 1.97, 95% CI 1.04 to 3.70) were associated with significantly increased odds of remission of all warts compared to podophyllin. We performed two further analyses. When subjects defaulting from follow up were assumed to have been cured odds of remission of all warts were also significantly increased both for podophyllotoxin solution (OR 3.04, 95% CI 1.68 to 5.49) and for podophyllotoxin cream (OR 2.46, 95% CI 1.38 to 4.40). When subjects defaulting from follow up were assumed not to have been cured odds of remission of all warts were significantly increased for podophyllotoxin solution (OR 1.92, 95% CI 1.13 to 3.27), but not for podophyllotoxin cream (OR 1.17, 95% CI 0.69 to 2.00). Local side effects were seen in 24% of subjects, and recurrence of warts within 12 weeks of study entry in 43% of all initially cleared subjects, without statistically significant differences between the treatment groups. Direct, indirect, and total costs were similar across the three treatment groups. Podophyllotoxin solution was the most cost effective treatment, followed by podophyllotoxin cream, with podophyllin treatment being the least cost effective. CONCLUSIONS: Self treatment of anogenital warts with podophyllotoxin showed greater efficacy and cost effectiveness than clinic based treatment with podophyllin. PMID- 12902573 TI - Is HSV serology useful for the management of first episode genital herpes? AB - BACKGROUND: First episode genital herpes simplex virus (HSV) infections can be classified into three groups, primary genital herpes (no previous exposure to HSV), non-primary first episode (IgG antibody to HSV of the non-presenting type), and first episode with pre-existing IgG HSV antibodies. The use of IgM to classify first episode genital herpes has not been evaluated. OBJECTIVE: To evaluate the sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of HSV-1 and HSV-2 IgM antibodies for the diagnosis of first episode genital herpes, when compared with clinical diagnosis. METHODS: Patients with a first clinical episode of genital herpes were recruited. Sera were tested for IgG antibodies to HSV-2 using an indirect enzyme linked immunosorbent assay (ELISA). Equivocal results were resolved by western blot. HSV 1 IgG and IgM and HSV-2 IgM antibodies were detected using western blot. RESULTS: 157 patients were recruited. 31 were excluded (missing data or no detectable antibodies and negative viral isolation). Therefore, 126 patients were included in the analysis. 23 (18.3%) had primary genital herpes, 34 (27.0%) non-primary first episode, and 69 (54.8%) had pre-existing genital herpes. The specificity and PPV of HSV IgM was 100%; the sensitivity was 79% and the NPV 85%. CONCLUSION: IgM HSV serology may be useful in the management of some patients with first episode genital herpes and provide an indication of the source of infection. Drawbacks include the low sensitivity and NPV, lack of availability, IgM antibodies may occasionally be produced in response to recurrent infection and, finally, IgM antibodies may take up to 10 days to develop and last 7-10 days. PMID- 12902574 TI - Psychosocial impact of serological diagnosis of herpes simplex virus type 2: a qualitative assessment. AB - OBJECTIVES: To assess the emotional and psychosocial responses to a serological diagnosis of HSV-2 infection in individuals without previous history of genital herpes. METHODS: 24 individuals who had a positive HSV-2 serology by western blot and no clinical history of disease were recruited from four clinics (sexually transmitted disease, maternal and infant care, family medicine, and virology research) over a 10 month period. In-depth qualitative interviews were conducted to elicit an individual's responses to the HSV-2 diagnosis. RESULTS: Three categories of themes were identified from the interviews. Short term emotional responses included surprise, denial, confusion, distress, sadness, disappointment, and relief to know. Short term psychosocial responses included fear of telling sex partners, anger at the source partner, guilt about acquiring or transmitting, and concern about transmitting to a child. Perceived ongoing responses included fear of telling future partners, concern about transmitting to a sex partner, feeling sexually undesirable, feeling socially stigmatised, feeling like "damaged goods," sex avoidance due to social responsibility, fear of transmitting to a newborn, and relationship concerns relating to the diagnosis. CONCLUSIONS: Individuals exhibit strong emotional and psychosocial responses to a serological diagnosis of HSV-2 infection. Many of the negative responses may be time limited and influenced by factors that are potentially amenable to counselling. PMID- 12902577 TI - An outbreak no longer: factors contributing to the return of syphilis in Greater Manchester. AB - BACKGROUND: In the past few years, increased diagnoses of syphilis have been reported in cities around Britain and Europe. Enhanced surveillance of cases began in 1999 to identify the epidemiology of this increase in Greater Manchester. METHODS: Information was collected on all cases of syphilis newly diagnosed in genitourinary medicine (GUM) clinics in Greater Manchester between January 1999 and November 2002. The data collected included demographic information and information about other sexually transmitted infections, sexual behaviour, perception of risk of infection, and awareness of syphilis transmission. RESULTS: The majority of cases identified were white homosexual men resident in Greater Manchester. Of the 414 cases diagnosed, 74% had either a primary or secondary stage of syphilis infection and 37% of cases were HIV positive. High numbers of individuals practised unprotected oral sex despite good awareness of the risk of infection with syphilis. There is evidence that the way people are meeting sexual contacts is changing, with increasing numbers meeting most of their partners through the internet. CONCLUSIONS: These findings have implications for targeting interventions. The provision of rapid diagnostic and treatment services is likely to be key for the control of syphilis and potentially of subsequent increases in HIV in the region. PMID- 12902576 TI - Seroepidemiological study of herpes simplex virus types 1 and 2 in Brazil, Estonia, India, Morocco, and Sri Lanka. AB - BACKGROUND: The association between herpes simplex virus type 2 (HSV-2) and human immunodeficiency virus (HIV) and the development of HSV vaccines have increased interest in the study of HSV epidemiology. OBJECTIVES: To estimate the age and sex specific seroprevalence of HSV-1 and HSV-2 infections in selected populations in Brazil, Estonia, India, Morocco, and Sri Lanka. METHODS: Serum samples were collected from various populations including children, antenatal clinic attenders, blood donors, hospital inpatients, and HIV sentinel surveillance groups. STD clinic attenders were enrolled in Sri Lanka, male military personnel in Morocco. Sera were tested using a common algorithm by type specific HSV-1 and HSV-2 antibody assay. RESULTS: 13,986 samples were tested, 45.0% from adult females, 32.7% from adult males, and 22.3% from children. The prevalence of HSV-1 varied by site ranging from 78.5%-93.6% in adult males and from 75.5%-97.8% in adult females. In all countries HSV-1 seroprevalence increased significantly with age (p<0.001) in both men and women. The prevalence of HSV-2 infection varied between sites. Brazil had the highest age specific rates of infection for both men and women, followed by Sri Lanka for men and Estonia for women, the lowest rates being found in Estonia for men and India for women. In all countries, HSV-2 seroprevalence increased significantly with age (p<0.01) and adult females had higher rates of infection than adult males by age of infection. CONCLUSIONS: HSV 1 and HSV-2 seroprevalence was consistently higher in women than men, particularly for HSV-2. Population based data on HSV-1 and HSV-2 will be useful for designing potential HSV-2 vaccination strategies and for focusing prevention efforts for HSV-1 and HSV-2 infection. PMID- 12902578 TI - Expression of the cytolethal distending toxin in a geographically diverse collection of Haemophilus ducreyi clinical isolates. AB - OBJECTIVE: To screen a collection of isolates of Haemophilus ducreyi for expression of the cytolethal distending toxin (CDT). METHODS: 45 clinical isolates of H ducreyi were screened for cytotoxic activity by examining the effect of culture supernatants on Hela cells. Expression was confirmed using immunoblotting with CDT specific monoclonal antibodies and the presence of the cdt genes determined by amplification of the cdt genes in a multiplex polymerase chain assay. RESULTS: Of the 45 clinical isolates, six isolates from differing geographical origins did not demonstrate cytotoxic activity. Expression of CDT was also not detected in these six isolates using immunoblotting and the genes cdtA, cdtB, and cdtC were not amplified using PCR. The remaining isolates demonstrated cytotoxic activity, expressed the CDT proteins, and the presence of the cdt genes was confirmed. CONCLUSIONS: CDT is considered a virulence factor of H ducreyi but was found to be absent in 13% of isolates from different geographical origins. PMID- 12902579 TI - Detection of varicella zoster virus in genital specimens using a multiplex polymerase chain reaction. AB - OBJECTIVE: To compare the relative proportions of varicella zoster virus (VZV) and herpes simplex viruses in specimens obtained from the genital lesions of adults presenting with presumed genital herpes infection. METHODS: Swabs of genital lesions from 6210 patients attending general practices, infectious diseases clinics within hospitals, or sexual health centres for treatment of their genital lesions were tested using polymerase chain reaction (PCR) technology. The multiplexed PCR was capable of detecting herpes simplex virus types 1 and 2 (HSV-1, HSV-2), VZV, and cytomegalovirus in a single sample. RESULTS: A total of 2225 patients had viruses detected by PCR. HSV-1 was detected in 36%, HSV-2 in 61%, and VZV in 2.9% of PCR positive samples. Of the 65 patients with VZV genital infection, many were thought to have HSV infection before laboratory testing. CONCLUSIONS: The finding of VZV in nearly 3% of virus positive genital specimens demonstrates that this virus needs to be considered as a differential diagnosis for genital herpetic lesions. Advice provided to patients with VZV genital infection regarding the source of infection, likelihood of recurrence, and potential for transmission of the virus will be different from that given to patients with HSV infection. PMID- 12902580 TI - A national survey of genitourinary medicine clinic attenders provides little evidence of sexual transmission of hepatitis C virus infection. AB - OBJECTIVE: To determine the prevalence and genetic diversity of hepatitis C virus in genitourinary medicine clinic attenders and to assess the extent of sexual transmission of the virus. METHODS: A cross sectional, unlinked, anonymous survey in 14 genitourinary medicine clinics situated in England, Wales, and Northern Ireland. Serum specimens from genitourinary medicine clinic attenders, retained as part of the Unlinked Anonymous Prevalence Monitoring Programme (UAPMP) serum archive, were tested in small pools, for the presence of antibody to hepatitis C virus (anti-HCV). The main outcome measures were prevalence of antibodies to hepatitis C virus and identification of hepatitis C virus genotypes. RESULTS: Testing of 17,586 specimens from 1995 showed an adjusted prevalence of anti-HCV in genitourinary medicine clinic attenders of 1.03% (95% CI: 0.89 to 1.16) overall and 0.65% (95% CI: 0.51 to 0.78) among those who did not report injecting drug use. Prevalence in injecting drug users attending genitourinary medicine clinics was 36.9% in both 1995 and 1996. Heterosexual injecting drug users had a higher prevalence of anti-HCV than homosexual/bisexual injectors. The most common hepatitis C genotypes were types 3a and 1a. There was a high degree of concordance between genotype and serotype. CONCLUSIONS: The low prevalence of anti-HCV in genitourinary medicine clinic attenders who deny injecting drugs suggests that the majority of hepatitis C infections have been acquired in adult life, mostly by injecting drug use, and that the hepatitis C virus is rarely transmitted sexually. The use of needle exchanges may explain the relatively low prevalence observed in the injecting drug users. PMID- 12902582 TI - Variations of HIV and STI prevalences within communities neighbouring new goldmines in Tanzania: importance for intervention design. AB - OBJECTIVES: To measure the prevalence of HIV and other STIs in communities neighbouring new large scale gold mines in northern Tanzania in order to inform the design of a targeted HIV/STI intervention programme. METHODS: Cross sectional surveys were conducted in adults aged 16-54 years from different sectors of communities neighbouring two newly opened, large scale gold mines near Lake Victoria. Mine workers, men, women, and female food and recreational facility workers (FRFW) from the community were randomly selected for interview and HIV and STI testing. RESULTS: 207 male Tanzanian mine workers, 206 FRFW, 202 other male and 205 female community members were enrolled. Overall, 42% of FRFW were HIV positive, compared to 6% of male mine workers, and 16% and 18% of other community men and women respectively. HIV prevalence in FRFW was significantly associated with alcohol consumption (adjusted odds ratio (aOR) = 2.5, 95% confidence interval (CI) 1.1 to 5.5), past or present syphilis (TPPA+) (aOR = 2.7, 95% CI 1.4 to 5.1) and single status (aOR = 3.8, 95% CI 1.2 to 11.9). Among FRFW, 24% had active syphilis (RPR+, TPPA+), 9% Chlamydia trachomatis, and 4% Neisseria gonorrhoeae. Overall, 50% of FRFW and 50% of community men never used condoms during sex, and 55% mineworkers, 61% male, and 20% female community members reported receiving/giving payment for sex during the previous year. CONCLUSIONS: There is a high prevalence of HIV and other STIs in communities around new goldmines in Tanzania, especially in FRFW. HIV and STI prevalence in the mining workforce is still relatively low, but high risk sexual behaviour is reported by all adult subgroups surveyed in this study. Programmes focusing on HIV/STI prevention, with targeted interventions for high risk women such as FRFW, will be extremely important in such high transmission communities where there is substantial recent in-migration of men and women seeking work. Such programmes have recently been initiated by a private/public/NGO partnership. PMID- 12902583 TI - Chlamydial infection: an accurate model for opportunistic screening in general practice. AB - OBJECTIVES: To estimate the prevalence of Chlamydia trachomatis in women in general practice and to assess risk factors associated with infection. METHODS: The study was carried out in 2001-2 in different general practices in Antwerp, Belgium. Sexually active women, visiting their general practitioner for routine gynaecological care (mostly pill prescription or PAP smear), were offered opportunistic screening for chlamydia. 787 participants aged 15-40 delivered a self taken vaginal sample and filled in a questionnaire which included questions on demographic variables, urogenital symptoms, sexual history, and sexual behaviour. Samples were tested for presence of chlamydial DNA by means of a ligase chain reaction (LCR) assay, and positives were confirmed by two other amplification assays (PCR and SDA). RESULTS: Overall prevalence was 5.0% (95% CI: 3.5 to 6.5). Determinants of infection in logistic regression analysis were age 18-27 years, >1 partner in the past year, no use of contraceptives, frequent postcoital bleeding, having a symptomatic partner, painful micturition, and living in the inner city. The area under the ROC curve in the full model was 0.88. Selective screening based on a combination of the five first determinants detects 92.3% of infections in this sample; 37.5% of the population would need to be screened. CONCLUSION: Targeted screening for chlamydial infection is possible, even in a heterogeneous group of general practice attendants. Implementing this model would require considerable communication skills from healthcare providers. PMID- 12902584 TI - Tetracycline treatment does not eradicate Mycoplasma genitalium. AB - OBJECTIVES: To study the treatment efficacy of tetracyclines and azithromycin in Mycoplasma genitalium positive patients attending an STD clinic. METHODS: All M genitalium positive patients (34 men and 26 women) attending an STD clinic during a 6 month period were treated with antibiotics. All patients known to be partners of M genitalium positive patients and those who were M genitalium positive, but not initially treated, were treated with azithromycin. Patients with urethritis and/or cervicitis were treated with tetracyclines before their M genitalium status was known. RESULTS: 10 of 14 women (71%) and 10 of 16 men (63%) treated with tetracyclines were M genitalium positive at follow up, whereas all patients treated with azithromycin (16 men and 20 women) were M genitalium negative, at the 4 week follow up visit. CONCLUSIONS: These results suggest that tetracyclines are not sufficient to eradicate M genitalium. Randomised controlled treatment trials are urgently needed. PMID- 12902585 TI - Awareness of human papillomavirus among women attending a well woman clinic. AB - OBJECTIVES: To assess the level and accuracy of public understanding of human papillomavirus (HPV) in the United Kingdom. METHODS: Women attending a well woman clinic were asked to complete a questionnaire assessing HPV awareness and specific knowledge about the virus. RESULTS: Questionnaires were completed by 1032 women, of whom 30% had heard of HPV. Older women, non-smokers, and those with a history of candida, genital warts, or an abnormal smear result were more likely to have heard of HPV. Even among those who had heard of HPV, knowledge was generally poor, and fewer than half were aware of the link with cervical cancer. There was also confusion about whether condoms or oral contraceptives could protect against HPV infection. CONCLUSIONS: In this relatively well educated sample, awareness and knowledge of HPV were poor. Public education is urgently needed so that women participating in cervical cancer screening are fully informed about the meaning of their results, especially if HPV testing is soon to be introduced. PMID- 12902586 TI - Syphilis prevalence has rapidly decreased in South Korea. AB - OBJECTIVES: To determine the current prevalence of syphilis in the general population of South Korea and its trend over the past 20 years. METHODS: 15,402 adults, who all were aged over 20 (6888 blood donors, 4951 physical examinees, and 3563 pregnant women) were examined from January 1999 to December 2000. All subjects were screened using a VDRL test, and positive results were confirmed by a FTA-ABS test. The prevalence was compared with the results of previous surveys in similar population groups performed by same authors. RESULTS: The prevalence of syphilis in South Korea was found to be 0.2%. Although has been consistent decreasing trend since 1977 (p<0.0001), there is no statistical difference between 1995 and 2000 (p=0.6992). CONCLUSIONS: The prevalence of syphilis has decreased significantly since 1977. However, it has not reduced much since 1995, and it seems that the prevalence has reached a plateau. Until now, not all syphilis patients were registered and followed up by the public health centres because there were so many syphilis patients. However, as this survey shows, the number of syphilis patients has reduced a great deal and it is believed that a more thorough management strategy including all newly diagnosed patients is necessary in order to further reduce the prevalence of syphilis in South Korea. PMID- 12902587 TI - Effects of two telephone survey methods on the level of reported risk behaviours. AB - OBJECTIVE: Reporting bias due to social desirability is an important consideration in carrying out surveys on sensitive issues. The study compared the frequency of self reported sensitive behaviours and response rates between the conventional "telephone interviewer method" (TIM) and a combined interviewer and computerised data capturing method (telephone interviewer and computerised questionnaire method, or TICQM). METHODS: A total of 580 males and 582 females were recruited and randomly assigned to either of the two methods in a cross sectional study. The overall response rate was about 51.6%. RESULTS: While the two methods both had high completion and low item non-response rates, the TICQM respondents reported higher frequencies of sensitive risk behaviours. Sexually active female respondents interviewed by the TICQM were more likely to report that their sex partners were their steady boyfriend, instead of their husband; and were also more likely to admit that they had had "one night stand" experiences or had undergone an HIV antibody test, when compared with their counterparts in the TIM group. Similar contrasts were observed for sexually active male respondents, that the TICQM group were more likely to report that they had had sexual intercourse with female sex workers or non-regular sex partners. Sex differences in the strength of association were observed between some studied behaviours (for example, HIV testing and substance abuse) and modes of data collection. CONCLUSION: The choice of data collection method has a significant impact on the results of sensitive studies; special attention should be given to designing the study and interpreting the results. PMID- 12902589 TI - Predictors of the prevalence of bacterial STI among young disadvantaged Indigenous people in north Queensland, Australia. AB - OBJECTIVE: To identify sexually transmitted infections in rural and remote Indigenous communities in north eastern Australia and examine factors that may influence prevalence. METHODS: A cross sectional survey of 26 Aboriginal and Torres Strait Islander communities in northern Queensland was carried out. 3313 people (2862 Indigenous) aged 15 years and over resident in participating communities during the period March 1998 to December 2000. The main outcome measures were community and population prevalence of chlamydia and gonorrhoea and independently associated risk factors. RESULTS: A total of 238 cases of chlamydia, 66 cases of gonorrhoea, and 37 cases of co-infection were detected among Indigenous participants. Prevalence of chlamydia and/or gonorrhoea ranged from 23.0% among 15-19 year olds to 3.5% among those 40 years and older. In the adjusted analysis younger age, female sex, lower socioeconomic status, the use of alcohol and tobacco, and the structure of community health services were independently associated with a higher prevalence of bacterial STI. CONCLUSION: This study highlights the need for improved STI control in north Queensland Indigenous communities through strategies to improve the reach and accessibility of primary healthcare services. PMID- 12902591 TI - Primary human parvovirus B19 infection in an HIV infected patient on highly active antiretroviral therapy. AB - Adults developing primary human parvovirus B19 (B19) infection may present with arthralgia, fever, and maculopapular rash. Recovery is linked to the development of specific neutralising antibodies. In immunosuppressed patients, including those with HIV infection, such humoral responses are impaired and severe chronic bone marrow suppression and arthritis may occur. PMID- 12902592 TI - Adult respiratory distress syndrome as a severe immune reconstitution disease following the commencement of highly active antiretroviral therapy. AB - We report a patient who developed adult respiratory distress syndrome (ARDS) secondary to Mycobacterium tuberculosis as an immune reconstitution disease. This complication occurred 14 days after the commencement of highly active antiretroviral therapy (HAART) for advanced HIV infection. The case emphasises that immune reconstitution can be an extremely aggressive complication of HAART. PMID- 12902596 TI - Emerging of dual AIDS associated neoplastic diseases in the era of highly active antiretroviral therapy. PMID- 12902597 TI - Pharyngeal gonorrhoea: the forgotten reservoir. PMID- 12902594 TI - Lactic acidosis in HIV infected patients: a systematic review of published cases. AB - OBJECTIVE: To describe the clinical, epidemiological, and biochemical characteristics of published cases of lactic acidosis (LA) and to generate hypotheses concerning risk factors associated with this complication. METHODS: Systematic review of cases reported in the medical literature. RESULTS: 217 published cases were identified, 90 of which fulfilled the study definition and had sufficient individual data on potential risk factors to be included. The 90 patients had a mean age of 40.1 years (range 16-69) and 53% were female. All 90 patients were taking nucleoside reverse transcriptase inhibitors (NRTI) at the time of the episode. Among the 83 patients with details of their antiretroviral therapy (ART) regimen 51 patients were taking stavudine, 29 zidovudine, 27 didanosine, and 25 lamivudine. Around 50% of the patients had abdominal pain, nausea, or vomiting. Hepatic steatosis was consistently reported (53/90) and in 36 (68%) there was histological evidence. The case fatality rate was 48%. Six cases were rechallenged with NRTI and three developed a further LA episode. Using data on the numbers of HIV infected individuals receiving care in the United States, we estimate that the risk of LA could be 2.5 times higher for women than men. CONCLUSIONS: NRTI use and female sex appear to be risk factors for the development of LA. What other factors are involved is still not clear but might include duration of NRTI therapy, specific drug use, and genetic predisposition. A case-control study is needed to better define risk factors for severe LA. PMID- 12902598 TI - Improving response rates for self collected urine samples. PMID- 12902599 TI - Impact of the Sexually Transmitted Infections Foundation course on the knowledge of family planning nurses and doctors. PMID- 12902600 TI - Which factors affect access to STD care? A comparison of a hospital based clinic and an outreach service. PMID- 12902601 TI - Clearance of HPV infection in middle aged men and women after 9 years' follow up. PMID- 12902602 TI - Follow up of patients who have been recently sexually assaulted. PMID- 12902603 TI - Patients lost to follow up: experience of an HIV clinic. PMID- 12902604 TI - Are all genital Chlamydia trachomatis infections pathogenic? A study in men. PMID- 12902605 TI - Interactive continuing medical education (CME) and its influence on the working practices of genitourinary clinicians. PMID- 12902606 TI - Syphilis specific antibodies in newborn infants in Lower Saxony, Germany 1993 2001. PMID- 12902607 TI - Lower motor neuron syndrome and HIV infection. PMID- 12902608 TI - Recent pilot studies of chlamydia screening. PMID- 12902609 TI - Chlamydia testing before termination of pregnancy. PMID- 12902610 TI - Rural sex work in Cambodia: work characteristics, risk behaviours, HIV, and syphilis. AB - OBJECTIVE: To identify prevalence and risks factors for syphilis and HIV among rural female sex workers (FSWs) in Cambodia and to describe differences between rural and urban FSWs. METHODS: Interviews and sera were collected from 114 FSWs and tested for HIV using the Serodia-HIV test and positives confirmed with the enzyme linked immunosorbent assay. Syphilis was tested for with the rapid plasma reagin with passive particle agglutination test for detection of antibody of Treponema pallidum. Study data were merged with data from a study of urban FSWs from Phnom Penh that applied similar questionnaires and sampling design to compare STI prevalence and behaviours. RESULTS: 42% of rural FSWs were HIV positive; 22% had past or current syphilis. In multivariate models HIV was significantly associated with age > or = 25 (OR = 6.1 95% CI: 1.0 to 36.6), a non commercial partner in the past year (OR= 0.33, 95% CI: 0.11 to 0.93), and prevalence of past or current syphilis (OR = 2.9, 95% CI: 1.0 to 8.8). There was significantly higher active syphilis (14% v 4%), older mean age (25 v 21), fewer daily clients (2 v 5), lower monthly income (61 US dollars v 174 US dollars), and longer duration of sex work (2.3 years v 1.4 years) among rural than among urban FSWs. CONCLUSIONS: These findings reveal a high burden of HIV and syphilis among FSWs in rural Cambodia. As FSWs age and become infected with STI/HIV they may move out of cities into less competitive but less savvy markets; their high mobility may contribute to the expansion of the HIV epidemic into rural Cambodia and lower risk populations. PMID- 12902611 TI - Descending thoracic aortic diseases: stent-graft repair. AB - PURPOSE: To evaluate endovascular treatment of descending thoracic aorta with commercially available self-expanding stent-grafts. MATERIALS AND METHODS: Seventy patients with aortic dissection, intramural hemorrhage, degenerative and posttraumatic aneurysm, penetrating atherosclerotic ulcer, and pseudoaneurysm underwent endovascular treatment. Eleven patients had impending rupture and were treated on an emergency basis. Stent-grafts were customized or selected on the basis of spiral computed tomographic (CT) or magnetic resonance (MR) imaging measurements. Preprocedure diagnostic angiography was performed in patients with aortic dissection and in other selected patients. All procedures were performed in an operating room and monitored with digital subtraction angiography (DSA) and transesophageal echocardiography (TEE). Follow-up was at 1, 3, 6, and 12 months after treatment and yearly thereafter. RESULTS: Stent positioning was technically successful in 68 cases. At DSA and TEE, complete aneurysm or false-lumen exclusion was achieved in 66 (97%) cases. No intraoperative mortality or complications occurred. In-hospital complications included transient monoparesis (one patient) and extension of dissection into ascending aorta (one patient) that was repaired surgically. Early endoleak was observed in five (7%) patients: In three (type 2), endoleak resolved spontaneously; in one (type 1), it was persistent; and in one (type 1), treatment was converted to surgery. At long term, one (1%) patient died of aortic rupture; another, of respiratory insufficiency. Five (7%) late endoleak (type 1, one caused by migration of the stent) cases were observed. In three (4%), endovascular treatment was successful; in two (3%), surgery was performed. In one patient with persistent postimplantation syndrome, treatment was converted to surgery after successful aneurysm sealing. Procedure failure (ie, aortic disease-related mortality or conversion to surgery) occurred in six (9%) patients. CONCLUSION: Endovascular stent-graft repair is less invasive in patients with chronic and acute descending thoracic aortic aneurysm and dissection. PMID- 12902615 TI - Participation of the mineralocorticoid receptor in cardiac and vascular remodeling. AB - The mineralocorticoid receptor mediates the effects of aldosterone. In addition to its influence on fluid and electrolyte, aldosterone plays a role in cardiac hypertrophy, tissue fibrosis, and modulation of vascular reactivity. However, the intracellular signaling mechanisms induced by aldosterone are not fully understood. Aldosterone causes slow transcriptional and rapid non-transcriptional effects. The latter include changes in intracellular calcium, c-Jun N-terminal kinase (JNK) and extracellularly regulated kinase (ERK) phosphorylation. In addition, autocrine loops involving other molecules such as the epidermal growth factor (EGF) receptor are also important in aldosterone-induced cellular mechanisms. This short review covers in vitro and in vivo studies that have investigated the pathophysiological role of aldosterone and the mineralocorticoid receptor in the cardiovascular system. PMID- 12902614 TI - Effect of donor/recipient body weight mismatch on patient and graft outcome in living-donor kidney transplantation. AB - BACKGROUND/AIMS: There have been conflicting reports showing that kidneys from small donors may be at risk for graft loss if they are transplanted into large recipients. The aim of this work was to examine the donor/recipient body weight ratio (D/RBWR) on patient and graft outcome. METHODS: During the period from January 1990 to January 2002, 856 kidney transplants were performed. Of these, 776 kidney transplant recipients were selected after exclusion of pediatric, second transplant patients and those with a body mass index of 35. All patients achieved a minimum follow-up of 1-year. According to D/RBWR, patients were divided into 3 groups: low (0.9), medium (0.91-1.2) and high (1.2). Data were collected on graft function, acute and chronic rejection, post-transplant complications, and 1- and 5-year graft and patient survival. RESULTS: There was a statistically significant increase in the incidence of chronic rejection, post transplant hypertension and diabetes mellitus in the low group. The incidence and frequency of acute rejection episodes were nearly the same in the 3 groups. Graft function, estimated by serum creatinine at 1 year, was significantly lower in the low group. The 5-year graft and patient survival was 71, 80, 88 and 81, 85 and 92%, in the low, medium and high groups, respectively. CONCLUSIONS: We conclude that a low D/RBWR may contribute to inferior long-term renal allograft survival. The hyperfiltration hypothesis due to low nephron mass in the low D/RBWR group may explain these findings. PMID- 12902616 TI - Renal and cardiovascular responses to water immersion in essential hypertension: is there a role for the opioidergic system? AB - Our study aimed at elucidating the effects of acute central hypervolemia induced by water immersion (WI) on renal hemodynamics, hormonal responses and on cardiovascular control in hypertensive patients, as well as at evaluating the possible role of the opioidergic system (OS) in determining these effects. Thirteen essential hypertensives were studied for 2 h before and for 2 h during WI. This was done twice, without and with i.v. injection of the OS antagonist naloxone. Before and during WI alone, glomerular filtration rate (GFR), effective renal plasma flow (ERPF), renal vascular resistance (RVR), mean arterial pressure (MAP), pulse interval (PI), spontaneous baroreflex sensitivity (BRS), Low frequency to High frequency (LF/HF) ratio in PI spectra, hematocrit, urinary sodium excretion, plasma renin activity (PRA) and aldosterone (PA) were assessed. Based on their response to WI, hypertensives were subdivided into two groups: ERPF+ (n = 7) in whom WI increased ERPF, and ERPF- (n = 6) in whom WI reduced ERPF. ERPF+ displayed a higher BRS than ERPF- at baseline and during WI. A suppression of PRA and PA and an increase in MAP and urinary sodium excretion were found in both groups. In ERPF+ naloxone caused RVR and MAP to increase during WI and this response was associated with a blockade of the increase in ERPF in this group, while BRS and natriuresis were unchanged. In ERPF- naloxone did not affect WI-induced MAP, ERPF, RVR and BRS changes, while it blunted sodium excretion. Our data provide the first evidence of a differentiate renal hemodynamic response to WI in hypertension; they also suggest that while OS may significantly potentiate the renal vasodilatory response to WI in ERPF+, it does not affect the natriuretic response nor the changes in systemic cardiovascular regulation induced by central hypervolemia. PMID- 12902617 TI - Integrins: sensors of extracellular matrix and modulators of cell function. AB - Integrins are a large family of transmembrane receptors for extracellular matrix (ECM) molecules. They play a critical role in organ morphogenesis, physiology and pathology, as they can modulate and control different cell functions, including adhesion, shape, polarity, growth, differentiation and motility. Integrins interact with ECM components via their extracellular domains, while their cytoplasmic domains play a pivotal role in mediating integrin-dependent cellular functions. The integrin cytoplasmic tails interact with the cytoskeleton, signaling molecules and other cellular proteins, resulting in regulation of many biological functions. In this review, we will mainly describe the role of integrins in regulating cell motility and discuss some new paradigms in integrin biology that may impact upon nephrology with respect to renal development and renal functions during both physiological and pathological events. PMID- 12902618 TI - Podocyte changes after induction of acute albuminuria in mice by anti aminopeptidase A mAb. AB - Administration of a specific combination of anti-aminopeptidase A (APA) mAb (ASD 37/41) in mice induces an acute albuminuria which is independent of angiotensin II, a well-known substrate of APA. In the present experiments, we examined whether binding of the mAb initiated changes in the podocytic expression of cytoskeleton (-associated), adhesion and slit-diaphragm proteins in relation to the time course of albuminuria. In addition, we measured ultrastructurally the extent of foot process retraction (the number of foot processes per microm GBM) and the width of the slit pore between the podocytes by morphometric methods. An injection of the mAb combination ASD-37/41 induced a massive but transient albuminuria that started at 6 h, and peaked at 8 h, after which it declined. However, even at day 7 after injection of the mAbs some albuminuria was present. Injection of the combination ASD-3/41 or saline did not induce an albuminuria. Notably, we observed changes in the staining of CD2AP and podocin, two slit-pore associated proteins that coincided with the start of the albuminuria. Nephrin staining was reduced and podocytic actin staining became more granular only at a time albuminuria was declining (24 h). The number of foot processes per microm GBM was already decreased at 4 h with a further reduction thereafter. The width of the slit pore was unchanged at the time of peak albuminuria and gradually decreased thereafter. At day 7, podocytic foot process effacement was even more prominent although albuminuria was only slightly abnormal. Expression of CD2AP was still granular. We observed however a change toward normal in the expression of podocin. Injection of saline or ASD-3/41 had no effect on the expression of podocytic proteins, the number of foot processes or width of the slit pore. Our data show that the onset of albuminuria in the anti-APA model is related to alterations in CD2AP and podocin, proteins that are important for maintaining slit-diaphragm structure and podocytic function. Extended studies at day 7 demonstrated uncoupling of albuminuria, podocytic foot process effacement and CD2AP staining. Changes in podocin more closely paralleled changes in albuminuria. PMID- 12902619 TI - HSP70 induced by hyperosmotic stress partially protects LLC-PK1 cells against nephrotoxic drugs. AB - Accumulation of HSP70 is related to the cytoprotection. It was evaluated whether hyperosmotic stress induces HSP70 accumulation in LLC-PK1 cells, and protects cells against toxicity provoked by cisplatin (Cis) and cyclosporine A (CyA). Cells were maintained in isosmotic (Iso) or hyperosmotic (H) culture medium for 24 h and then exposed to Cis or CyA for an additional period of 12 or 24 h (groups H+Cis and H+CyA). The H medium did not induce cell death and increased both HSP70 mRNA and protein levels, suggesting a role in cell adaptation to H condition. H medium produced partial cytoprotection against Cis and CyA compared with control cells. Despite the cytoprotection, there was a reduction in HSP70 mRNA and protein levels in H+Cis group. In contrast, the H+CyA group presented high levels of HSP70 mRNA and protein. The induction of HSP70 by H medium was associated with tolerance of LLC-PK1 cells against Cis and CyA cytotoxicity but this protection was induced by different mechanisms and depended on the characteristics of the drug used. PMID- 12902620 TI - Pericellular matrix formation by renal tubule epithelial cells in relation to crystal binding. AB - BACKGROUND/AIM: Retention of crystals in the kidney ultimately leads to renal stone formation. Hyaluronan (HA) has been identified as binding molecule for calcium oxalate monohydrate crystals. The association of high molecular mass (M(r)) HA with cell surface receptors such as CD44 gives rise to pericellular matrix (PCM) formation by many eukaryotic cells in culture. Here, we study the ability of several renal tubular cell lines to assemble PCMs and to synthesize high-M(r) HA during proliferation in relation to crystal retention. METHODS: PCM assembly by MDCK-I, MDCK-II, and LLC-PK1 cells was visualized by particle exclusion assay. Metabolic labeling studies were performed to estimate the cellular production of HA. The expression of CD44 and HA was studied using fluorescent probes, and crystal binding was quantified with radiolabeled calcium oxalate monohydrate. RESULTS: PCMs were formed, and HA was expressed by most MDCK I and some MDCK-II, but not by LLC-PK1 cells. All cell types expressed CD44 at their apical surface. MDCK-I and MDCK-II cells secreted, respectively, 14.7 +/- 1.6 and 0.5 +/- 0.2 pmol [3H]glucosamine incorporated in high-M(r) HA, whereas LLC-PK1 cells did not secrete HA. Streptomyces hyaluronidase treatment significantly decreased crystal binding (microg/cm2) to MDCK-I cells (from 8.6 +/ 0.4 to 3.9 +/- 0.9), but hardly to MDCK-II cells (from 10.2 +/- 0.2 to 9.6 +/- 0.1) or LLC-PK1 cells (from 10.2 +/- 0.8 to 9.9 +/- 0.3). CONCLUSIONS: There are various forms of crystal binding to renal tubular cells in culture. Crystal attachment to MDCK-I and some MDCK-II cells involves PCM assembly that requires high-M(r) HA synthesis. HA production and PCM formation do not play a role in crystal binding to LLC-PK1 and the majority of MDCK-II cells. It remains to be determined which form of binding is involved in renal stone disease. PMID- 12902621 TI - A Canadian cohort study of cognitive impairment and related dementias (ACCORD): study methods and baseline results. AB - The overall objective of the Canadian Collaborative Cohort of Related Dementias (ACCORD) study is to describe the diagnostic distribution, natural history and treatment outcomes of individuals referred from the community to dementia clinics in Canada. Between 1997 and 1999, an inception cohort of 1,136 subjects entered into this longitudinal study. At the baseline assessment, 10.9% of the subjects were classified as "not cognitively impaired" (NCI), 30.1% as "cognitively impaired not demented" (CIND), and 59% as demented. A subclassification of CIND included amnestic 25.1%, vascular cognitive impairment 18.1%, psychiatric 17.2%, neurologic 7.3%, medical/toxic metabolic 3.5%, mixed 7.6% and not specified 19.0%. The percentage of the cohort referred with dementia increased progressively each decade, while the proportions of CIND and NCI decreased. Within the dementia group, Alzheimer's disease accounted for 47.2% of the subjects, mixed dementias 33.7%, vascular dementia 8.7%, frontotemporal degenerations 5.4%, dementia with Lewy bodies 2.5%, and unclassifiable 1.8%. The ACCORD cohort will allow a detailed study of the longitudinal course of CIND, and the longer-term outcomes of both treated and untreated dementia subjects. PMID- 12902622 TI - A prospective analysis of risk factors for white matter disease in the brain stem: the Cardiovascular Health Study. AB - BACKGROUND AND PURPOSE: White matter disease (WMD) in the brain stem may be a predictor of poor clinical outcome, independent of WMD in the periventricular and subcortical areas of the brain. Many cardiovascular risk factors such as older age, hypertension, and smoking have been suggested as risk factors for WMD in the periventricular and subcortical areas of the brain. However, no epidemiologic study has examined the associations between cardiovascular risk factors and WMD in the brain stem. METHODS: A total of 789 participants, aged 65 years or older, from the Cardiovascular Health Study constituted the present study population. WMD, defined as hyperintensive lesions on magnetic resonance imaging (MRI), in the brain stem was measured in 1992/1993 and 1997/1998. RESULTS: Of the 789 participants, 212 (26.9%) had WMD in the brain stem in 1997/1998. In multivariate logistic regression analysis, the presence of WMD in the brain stem in 1997/1998 was significantly associated with several variables measured in 1992/1993: an increase by 5 years of age (OR = 1.51, 95% CI: 1.25-1.83), a 10-pack-years increase in smoking (OR = 1.12, 95% CI: 1.04-1.21), a 0.1-liter increase in first second forced expiratory volume (OR = 0.95, 95% CI: 0.92-0.99), a 1 micromol/l increase in fibrinogen level (OR = 1.13, 95% CI: 1.03-1.23), and MRI infarction (OR = 2.58, 95% CI: 1.78-3.74). Excluding those (n = 167) with WMD in the brain stem in 1992/1993, the pattern remained. Hypertension was not associated with WMD in the brain stem. CONCLUSIONS: Increased age, smoking, lower forced expiratory volume, increased fibrinogen level, and MRI infarction, but not hypertension, may be independent risk factors for WMD in the brain stem in older adults. PMID- 12902623 TI - Epidemiological and genetic studies of myotonic dystrophy type 1 in Taiwan. AB - To investigate the prevalence and genetic characteristics of myotonic dystrophy type 1 (DM1) in Taiwan, DM-suspected patients and their families identified during the period of 1990-2001 had their clinical records reevaluated and the CTG repeat sizes at the DM1 locus examined. A total of 96 subjects belonging to 26 families were identified as DM1 patients, which gave a minimal disease prevalence of 0.46/100,000 inhabitants. Clinical anticipation was frequently observed in affected families, even in some parent-child pairs with transmission contraction of the CTG repeat size. The inverse correlation between age at onset and CTG repeat length was significant only in patients with small expansions. In addition, a DM1 carrier with a childhood-onset son was found to have CTG length heterogeneity in the range of 40-50, indicating that premutation alleles could be unstable during gametogenesis as well as in somatic tissues. Our data demonstrated that DM1 is a rare disease in Taiwan and showed that transmission contraction of repeat size is more likely to occur in alleles with large repeats. PMID- 12902624 TI - Risk for relatives of patients with multiple sclerosis in central Sardinia, Italy. AB - Multiple sclerosis (MS) is a chronic, inflammatory, disabling disease of the central nervous system, known for its complex interplay between genetic and environmental factors. We used life table techniques to calculate age-adjusted recurrence risks for different categories of relatives of MS patients from Central Sardinia (Italy), a genetically homogeneous, stable population with a high degree of consanguinity. We included 313 probands and a total of 12,717 relatives in the analysis. The overall age-adjusted recurrence risk for relatives of MS probands is 1.90% [95% confidence interval (CI): 1.57-2.30]. The age adjusted recurrence risk in parents was 1.26% (95% CI 0.60-2.63), in children 2.33% (95% CI 0.09-5.56), in sibs 4.76% (95% CI 3.57-6.32), in second-degree relatives 0.72% (95% CI 0.42-1.22), and in third-degree relatives 1.79% (95% CI 1.27-2.51). The sex of the probands (male) and of the relatives (female), and the number of affected relatives in the family significantly increase the risk of MS in relatives. PMID- 12902625 TI - A case-control study on cigarette, alcohol, and coffee consumption preceding Parkinson's disease. AB - OBJECTIVE: To investigate the association between cigarette smoking, alcohol drinking, coffee consumption and Parkinson's disease (PD). METHODS: We selected subjects affected by idiopathic PD, with a Mini-Mental State Examination of > or =24, and controls matched 1 to 1 with cases by age (+/- 2 years) and sex. Controls were randomly selected from the resident list of the same municipality of residence of the cases. We assessed cigarette smoking, alcohol drinking, and coffee consumption preceding the onset of PD or the corresponding time for controls using a structured questionnaire, which also evaluated the duration and dose of exposure. Using conditional logistic regression analysis, we calculated adjusted OR and 95% CI. RESULTS: We interviewed 150 PD patients and 150 matched controls. Cigarette smoking (ever vs. never smokers OR = 0.66, 95% CI = 0.41 1.05, p = 0.08) did not show a statistically significant association with PD. We observed an inverse association between alcohol drinking (ever vs. never OR = 0.61, 95% CI = 0.39-0.97, p = 0.037) and coffee consumption (ever vs. never OR = 0.16, 95% CI 0.05-0.46, p = 0.0001) and PD. These associations remained significant after adjustment for other covariates: OR for ever vs. never alcohol consumption was 0.62 (95% CI = 0.43-0.89, p = 0.009) and that for coffee drinking 0.19 (95% CI = 0.07-0.52, p = 0.001). Heavy coffee consumption confirmed the inverse association between coffee and PD (more than 81 cup/year vs. none: OR = 0.20, 95% CI = 0.08-0.47, p < or = 0.0001). CONCLUSIONS: Consistent with previous studies, our results suggest an inverse association between coffee drinking, alcohol consumption and PD. The multiple inverse association observed may indicate a complex interaction between genetic and environmental factors. PMID- 12902627 TI - The incidence of Parkinson's disease in the Netherlands: results from a longitudinal general practice-based registration. PMID- 12902626 TI - Association between Parkinson's disease and exposure to pesticides in southwestern France. AB - A case-control study was performed in southwestern France in order to assess the relationship between pesticide exposure and Parkinson's disease (PD) in the elderly. During the period from 1997 to 1999, 84 cases were recruited together with 252 population-based controls. Experts in occupational health reviewed job codes and provided pesticide exposure levels, making it possible to calculate cumulated exposure lifelong for individuals. Environmental pesticide exposure was considered in relation to the place of residence. A positive association was found with occupational pesticide exposure (odds ratio = 2.2, 95% confidence interval 1.1-4.3) in conditional logistic multiple regression analysis taking into account age, sex, educational level and smoking; however, no clear dose relationship was found. Our results support the hypothesis of an association between occupational pesticide exposure and PD and point to the need to investigate the role of fungicides, for which toxicological hypotheses exist. PMID- 12902628 TI - Is endotoxin an environmental cause of Parkinson's disease? PMID- 12902630 TI - Antibodies: still important in anti-GBM disease. PMID- 12902631 TI - Sevelamer. AB - Hyperphosphataemia in patients with chronic renal failure has traditionally been treated with calcium- and aluminium-containing phosphate binders. This article briefly discusses the rationale and complications of these treatments and reviews sevelamer that is a novel non-calcium-, non-aluminium-containing phosphate binder and the evidence to support its use in clinical practice. PMID- 12902632 TI - The prognostic significance in Goodpasture's disease of specificity, titre and affinity of anti-glomerular-basement-membrane antibodies. AB - BACKGROUND: The nephrotoxic potential of anti-glomerular-basement-membrane (GBM) antibodies has been demonstrated in numerous animal experiments. However, it is not known to what extent the properties of circulating anti-GBM antibodies in human disease reflect the severity of the disease and predict the outcome. METHODS: Clinical data were collected for 79 Swedish patients for whom a positive result had previously been obtained with anti-GBM ELISA. In stored sera from the patients, we measured antibody concentration, specificity and affinity together with antineutrophil cytoplasmic antibodies and alpha(1)-antitrypsin phenotype. RESULTS: Six months after diagnosis, 27 (34%) were dead, 32 (41%) were on dialysis treatment and only 20 (25%) were alive with a functioning native kidney. The best predictor for renal survival was renal function at diagnosis. In patients who were not dialysis dependent at diagnosis however, renal survival was associated with a lower concentration of anti-GBM antibodies, a lower proportion of antibodies specific for the immunodominant epitope and the histological severity of the renal lesion. The only factor that correlated with patient survival was age. CONCLUSIONS: Immunochemical properties of autoantibodies do not affect patient survival in anti-GBM disease but seem to be a factor in renal survival in patients detected before renal damage is too advanced. PMID- 12902633 TI - Osteopenia in patients with glomerular diseases requiring long-term corticosteroid therapy. AB - BACKGROUND: Chronic corticosteroid (CS) use is associated with bone mass loss. METHODS: Bone mineral density (BMD) was assessed in 72 patients (25 males/47 premenopausal females) with glomerular diseases, primary (n = 35) or secondary to systemic lupus erythematosus (n = 37) with normal renal function, who were taking CS, as prednisone and/or methylprednisolone, in doses > or =7.5 mg/day, for a period of at least 6 months. Cumulative dose and duration of prior CS therapy, as well as biochemical parameters and other factors contributing to bone loss were evaluated. RESULTS: We found 37 (52%) patients with LOW BMD (29 with osteopenia and 8 with osteoporosis). The low BMD group presented a lower mean weight and body mass index (BMI) versus the normal BMD group (62 +/- 15 vs. 70 +/- 10 kg and 25 +/- 4 vs. 27 +/- 5, mean +/- SD, p < 0.05). The estimated calcium intake was lower than 400 mg/day in all patients with low BMD, and they had taken furosemide as a concomitant drug for a longer mean period of time when compared to normal BMD patients (30 +/- 29 vs. 16 +/- 27 months, p < 0.05). A higher mean number of pulses per patient and mean cumulative dose of methylprednisolone were observed in the low versus normal BMD group (7.7 +/- 4.0 vs. 5.6 +/- 4.0 pulses and 6.5 +/ 3.9 vs. 3.9 +/- 2.7 g, p < 0.05). CONCLUSIONS: These findings suggest a high frequency of osteopenia among young and premenopausal patients with glomerular diseases given long-term corticosteroid therapy. The lower BMI and calcium intake, as well as the concomitant furosemide use, might have contributed to such a bone loss. The higher number of pulse therapies leading to higher cumulative intravenous doses of corticosteroid mainly in lupus nephritis patients shows that pulse therapy may be deleterious to bone. PMID- 12902634 TI - Intermittent high-permeability hemofiltration modulates inflammatory response in septic patients with multiorgan failure. AB - BACKGROUND/AIM: Continuous venovenous hemofiltration with high-permeability hemofilters is a novel approach in the adjuvant therapy of septic patients. High permeability hemofilters are characterized by an increased pore size which facilitates the filtration of inflammatory mediators. The present study examines whether intermittent high-permeability hemofiltration has an immunomodulatory effect on polymorphonuclear leukocytes and mononuclear cells. METHODS: Twenty eight septic patients with acute renal failure were randomly allocated to either receive intermittent high-permeability or conventional hemofiltration. Intermittent high-permeability hemofiltration consisted of a daily 12-hour course of high-permeability hemofiltration alternated by conventional hemofiltration. For high-permeability hemofiltration, a newly developed high-flux polyamide membrane (P2SH) with a nominal cutoff point of 60 kD was used. For conventional hemofiltration a high-flux polyamide hemofilter (Polyflux 11S, cutoff point 30 kD) was used. RESULTS: The polymorphonuclear leukocyte phagocytosis activity before starting hemofiltration was almost double the rate of healthy controls in both groups (p < 0.001). The phagocytosis rate decreased significantly during the course of intermittent high-permeability hemofiltration (p < 0.05), whereas the values remained unchanged in the conventional hemofiltration group. Incubation of high-permeability filtrates with blood from healthy donors resulted in a significant induction of phagocytosis (p < 0.001), whereas conventional filtrates had no phagocytosis-stimulating effects. In addition, incubation of healthy-donor mononuclear cells with high-permeability but not conventional filtrates resulted in a significant tumor necrosis factor alpha release (p < 0.001). CONCLUSIONS: Intermittent high-permeability hemofiltration is a novel extracorporeal elimination modality which exhibits immunomodulatory effects on leukocytes, attenuating polymorphonuclear neutrophil phagocytosis. Further studies are necessary to elucidate whether these effects translate in a clinical improvement in patients suffering from sepsis. PMID- 12902635 TI - Integrins as unique receptors for vascular control. AB - Cells within the vascular wall connect their cytoskeleton to the extracellular matrix (ECM) through a family of cell surface receptors known as integrins. The ability of integrins to act as a link between the extracellular and intracellular environments allows transmission of inside-out and outside-in signals capable of modulating diverse vascular phenomena. In this review we summarize what is currently known about the involvement of integrins in the control of vascular tone, permeability and remodeling. We discuss the capacity of integrins to act as detectors of injury-generated molecules derived from ECM proteins, as well as the putative role of integrins as mechanosensors for shear and tension. Particular attention is given to the mechanisms responsible for linking integrins to the control of vascular tone, and we review the intracellular signaling pathways involved in effecting the vascular responses elicited by integrin activation. Finally, the involvement of integrins in vascular remodeling and vascular disease is analyzed. Considerable evidence strongly indicates that integrins are involved in both acute and chronic vascular control. Understanding the elements and the sequence of events linking integrins with vasoregulation is important for deciphering phenomena such as the pressure-dependent myogenic response, flow dependent changes in vascular diameter, and vascular remodeling as they occur in physiological and pathological conditions. Further understanding of the role of integrins in vascular control holds promise as new avenues for prophylactic and therapeutic manipulation of vascular phenomena. PMID- 12902636 TI - Adhesion of endothelial cells to NOV is mediated by the integrins alphavbeta3 and alpha5beta1. AB - NOV is a member of the CCN family of matricellular proteins. We have shown previously that NOV is strongly expressed by vascular smooth muscle cells (VSMCs) of the rat carotid artery. However, 7 days after injury, NOV expression is down regulated, except near the luminal surface of the developing intima, where it is strongly expressed. These data suggested that NOV might be involved in the regulation of endothelial cell adhesion. NOV promoted the adhesion of human umbilical vein endothelial cells (HUVECs), which was abolished by anti-NOV antibody. HUVEC adhesion to NOV required divalent cations and was inhibited by GRGDS peptide, implicating integrins in the adhesion mechanism. Monoclonal antibodies (mAbs) against alphavbeta3 inhibited adhesion of HUVECs to NOV, and NOV was shown to bind to alphavbeta3. Anti-alpha5beta1 mAbs also inhibited HUVEC adhesion to NOV, but adhesion via alpha5beta1 was mediated by fibronectin. HUVEC adhesion to NOV caused intracellular signalling, as evidenced by increased phosphotyrosine content of focal adhesion kinase. Together with evidence that NOV expression in a variety of tissues is restricted to blood vessels containing VSMCs, these data are consistent with a role for NOV in endothelial cell adhesion in vascular homeostasis and in the response to injury. PMID- 12902638 TI - Enhanced neuropeptide Y immunoreactivity and vasoconstriction in mesenteric small arteries from spontaneously hypertensive rats. AB - Enhanced sympathetic nerve activity is thought to play a role in the pathogenesis of hypertension. The purpose of the present study was to investigate the mechanisms underlying the enhanced vasocontractile response to perivascular stimulation of mesenteric arteries isolated from female spontaneously hypertensive rats (SHR). Innervation of mesenteric small arteries was evaluated by immunohistochemistry and confocal microscopy while functional studies were conducted in a microvascular myograph. The distribution of nerve terminals immunoreactive for tyrosine hydroxylase (TH) and neuropeptide Y (NPY) was similar in mesenteric small arteries from Wistar-Kyoto (WKY) and SHR rats. However, immunointensity of TH or NPY immunoreactivities were much higher in small arteries from SHR compared to WKY. Expressed as percentage of contractions elicited by 124 mM K(+), concentration-response curves for noradrenaline (NA) and NPY were shifted leftward in SHR compared with WKY rats. The combination of noradrenaline (1 microM) and NPY (10 nM) contracted mesenteric arteries from WKY and SHR to higher levels than compared to either contractile agent added alone. The NPY Y(1) receptor antagonist, BIBP 3226, inhibited these contractions with 87 +/- 0.7 and 80 +/- 1.3% (p < 0.05, n = 6) in arteries from WKY and SHR rats, respectively. In arteries incubated with the alpha(1)-adrenoceptor antagonist, prazosin, and preactivated with vasopressin, electrical field stimulation evoked contractions which were more pronounced in mesenteric arteries from SHR compared to WKY rats. BIBP 3226 partially inhibited these contractions. In vasopressin activated arteries BIBP 3226 caused rightward shifts of the concentration response curves for NPY in mesenteric arteries from SHR rats, but in addition it also abolished the maximal NPY contraction in arteries from WKY rats. In the presence of BIBP 3226, low concentrations (1 pM to 10 nM) of NPY caused relaxations in arteries from WKY, but not in segments from SHR rats. Mechanical removal of the endothelium abolished NPY relaxation in arteries from WKY. In arteries activated with vasopressin and exposed to either forskolin or sodium nitroprusside, the addition of NPY evoked contractions which were more pronounced in arteries from SHR compared to WKY arteries. The present study suggests that enhanced NPY content and vasoconstriction to NPY in arteries from hypertensive rats can contribute to the enhanced sympathetic nerve activity and vascular resistance in female hypertensive rats. Endothelial cell dysfunction as well as alterations in smooth muscle response to NPY seem to contribute to the enhanced vasoconstriction in arteries from hypertensive animals. PMID- 12902637 TI - Vessel- and vasoconstrictor-dependent role of rho/rho-kinase in renal microvascular tone. AB - We examined the role of Rho/Rho-kinase in renal afferent and efferent arteriolar tone induced by angiotensin (Ang) II, KCl and elevated renal arterial pressure (from 80 to 180 mm Hg), using isolated perfused rat hydronephrotic kidney. In the condition with no vasoconstrictor stimuli, Y-27632, a Rho-kinase inhibitor, dilated only afferent (from 11.6 +/- 0.4 to 14.1 +/- 0.5 microm) but not efferent arterioles (from 11.6 +/- 0.2 to 12.6 +/- 0.7 microm) at 10(-5) mol/l. During renal vasoconstriction by Ang II, Y-27632 restored the afferent arteriolar constriction (141 +/- 10% reversal at 10(-5) mol/l), whereas the ability of Y 27632 to inhibit the Ang II-induced efferent arteriolar constriction was diminished (73 +/- 7% reversal). A similar action was observed with fasudil, another Rho-kinase inhibitor. Furthermore, Y-27632 impaired myogenic afferent arteriolar constriction, with 117 +/- 17% inhibition at 10(-5) mol/l. The inhibition by Y-27632 of the myogenic vasoconstriction was almost the same as that of the Ang II-induced tone of this vessel type. However, Y-27632 had a modest effect on KCl-induced vasoconstriction of afferent arterioles. In conclusion, the present study demonstrates a predominant role of Rho/Rho-kinase in mediating the basal and Ang II-induced tone of afferent, but not efferent, arterioles. Furthermore, the role of Rho/Rho-kinase in afferent arteriolar constriction differs, with a substantial contribution to Ang II-induced and myogenic constriction but a minimal role in depolarization-induced constriction. Since Ang II-induced, KCl-induced and myogenic constriction of afferent arterioles require calcium entry through voltage-dependent calcium channels, the interaction between Rho/Rho-kinase and the calcium entry pathway may determine the afferent arteriolar tone induced by these stimuli. PMID- 12902639 TI - Beta-galactosidase-tagged adventitial myofibroblasts tracked to the neointima in healing rat vein grafts. AB - OBJECTIVE: Myofibroblasts are present transiently in normal healing wounds. However, they have been found to persist in the stroma of neoplasms, fibrotic conditions and other pathological settings. In rat vein grafts, we have observed the prolonged presence of myofibroblasts. Our aim was to determine the origin of myofibroblasts in vein grafts. METHODS: Epigastric vein to femoral artery grafts were microsurgically placed in male Lewis rats and harvested. Neointimal development, cellular death and proliferation, and cell phenotypes were analyzed using immunohistochemistry and light and electron microscopy. To follow cellular movement in the vessel wall, vein grafts were transfected with replication defective adenovirus containing the gene encoding beta-galactosidase (n = 50), and harvested at 1, 2, 3, 4, 5, 6, 7, 14 and 28 days. Grafts were analyzed after X-gal staining. RESULTS: Myofibroblasts were detected in the outer adventitia at 4 days, in the media at 1 week and in the developing neointima at 2 weeks. Cells tagged using adenoviral beta-galactosidase demonstrated adventitia to neointima cell migration. CONCLUSIONS: Although there may be other sources of myofibroblasts in this model, the adventitia has been shown to be an origin of myofibroblasts which subsequently migrate through the vessel wall to the neointima during graft remodeling and contribute to neointimal formation. PMID- 12902642 TI - The current pathobiology and chemotherapy of clonorchiasis. AB - This special issue is devoted to a review of the current state of knowledge of clonorchiasis. Sections deal with: the biology of Clonorchis sinensis; geographical distribution, with particular regard to Korea; epidemiology; pathogenesis; clinical features; diagnosis; prevention and control; chemotherapy. PMID- 12902640 TI - Cardiovascular protective effects of synthetic isoflavone derivatives in apolipoprotein e-deficient mice. AB - Dietary flavonoids are thought to protect against cardiovascular disease. We have studied the effects of bioactive isoflavone metabolites on hyperlipidemia, endothelial dysfunction and the development of atherosclerotic lesions in apolipoprotein E-deficient (apoE(0)) mice fed a Western high-fat diet. Supplementation with dihydrodaidzein (DiD), dehydroequol (DeE) (both 25 mg kg(-1) x day(-1)) and their combination (D/D; 12.5 mg kg(-1) x day(-1) for each) for 24 weeks reduced plasma high-density lipoprotein (HDL) and non-HDL cholesterol. D/D also reduced the triglyceride level. In the abdominal aorta of apoE(0) mice, these compounds significantly increased endothelial nitric oxide (NO)-mediated vasorelaxations induced by acetylcholine, but had a minor effect on relaxations induced by the NO donor S-nitroso-N-acetylpenicillamine. Isoflavone treatment for 24 weeks had no effect on the total area of atherosclerotic plaques in the whole aorta, but DeE reduced the plaque thickness in the aortic arch by 29%, although this did not reach statistical significance. The endothelial dysfunction in apoE(0) mice is associated with hyperlipidemia and increased vascular oxidative stress measured as increased superoxide production. Both isoflavones have superoxide-scavenging activities in vitro. We suggest that chronic supplementation with bioactive isoflavone metabolites may protect endothelial NO function in apoE(0) mice, through both lipid-lowering and antioxidant actions. PMID- 12902643 TI - Epidemiological Studies On Ascaris Lumbricoides Reinfection In Rural Communities In Korea: 1. The Relationship Between Prevalence And Monthly Reinfection Rate. AB - The epidemiological relationship between the current prevalence and monthly reinfection rate of Ascaris lumbricoides was observed in rural communities in Korea by measns of blanket mass chemotherapy and worm collection for measurement of the prevalence and reinfection rates. During the period from 1975 to 1980, a total of 4,466 inhabitants in 10 different localities were treated with 10 mg/kg of pyrantel pamoate and 2 days' whole stools were collected from 2,547 inhabitants. The stools were examined for the presence of expelled adult and/or young worms, which represent the prevalence and reinfection rates for past 2 months respectively. After then, the obtained rates were correlated each other applying the time-prevalence curve proposed by Hayashi. It was observed that the prevalence (overall worm positive rate) and worm burden per individual ranged by areas from 13.6 to 72.3% and 1.4~10.2 respectively. The calculated monthly reinfection rates (X) (from young worm positive rates) according to areas were in the range, 2.6~16.2%, and clearly correlated with the current prevalence (Y) under the equation, Y=1-(1-X)(7.2) where 7.2 is time in month. The equation means that after one time mass chemotherapy the period needed to attain equilibrium of prevalence again would be about 7~8 months. And it is inferred that the majority of reinfected worms in human host turn over every 7~8 months. PMID- 12902644 TI - Epidemiological Studies On Ascaris Lumbricoides Reinfection In Rural Communities In Korea II. Age-Specific Reinfection Rates And Familial Aggregation Of The Reinfected Cases. AB - Epidemiological studies on the reinfection pattern of Ascaris lumbricoides were undertaken by means of blanket mass chemotherapy and worm collection in a rural village in Korea, during 1977~1980. The study objectives were to determine the age(sex)-specific reinfection rate during 2, 4, 6 and 12 months through repeated mass chemotherapy with pyrantel pamoate, and to observe the familial aggregation tendency of the reinfected cases. The results obtained are as follows: 1. The age(sex)-reinfection curve revealed that the reinfection rate is much higher in younger individuals than in olders in all of 4 kinds of interval chemotherapy groups. The highest reinfection rate and the highest burden of reinfected worms were observed in preschool childen, followed by primary school students. Such fluctuation in the age-specific reinfection rates was more pronounced in males than in females. 2. There was noted a significant tendency of familial aggregation among the reinfected cases. It is suggested that reinfection occurs never randomly but preferably to the members of certain household families. From these reinfection analyses, it is inferred that the principal mode of A. lumbricoides transmission in the surveyed rural area is likely to be of 'dooryard type', in which case childen and certain family members are more preferably reinfected. It is also suggested that the preschool childen should be included in the primary targets of mass control programme. PMID- 12902645 TI - [Five Cases Of Diphyllobothrium Latum Infection] AB - Five cases of Diphyllobothrium latum infection were proved by collection of worms after treatment with bithionol, niclosamide or praziquantel, during 1975~1983. The patients were 4 males and one female aged 10-64 years residing in Seoul or Wando-gun, Chollanam-do, Korea. All of them remembered the history of eating several kinds of raw marine fish and 2 persons said they ate the raw flesh of perch, Lateolabrax japonicus. Three among them experienced abdominal pain, 2 natural discharge of a chain of segments, but none revealed any sign of anemia. The average egg size in stool varied 59-67 x 41~48 micrometer according to cases. The eggs were characterized by ovoid to elliptical shape, terminal operculum, and distinct abopercular protuberance. One naturally discharged segment (30 cm) and 4 complete strobilae (320~680 cm) with scolices obtained after treatments were examined. The morphological characters of proglottids such as rosette-form uterus with 3~6 loops, vaginal pore included in the cirrus sac, separated seminal vesicle from cirrus sac etc. were all compatible with D. latum. These are the 7th~11th cases of D. latum infection proved by worms in Korea. PMID- 12902646 TI - [Epidemiological Studies Of Clonorchis Sinensis In Mangyeong Riverside Areas In Korea] AB - In an attempt to clarify the epidemiological feature of C. sinensis in Mangyeong riverside area, the prevalence of clonorchiasis, infestation rate of the cercariae in Parafossarulus manchouricus, and detection rate of the metacercariae in fresh-water fishes were investigated from March 1 to September 30, 1983 at the upper, middle and lower reaches of the river. The results obtained were summarized as follows:Detection rate of C. sinensis egg among inhabitants was 8.2 % out of a total of 1,266 persons. but the differences in detection rates were not statistically signifcant among upper, middle and 1ower reaches. According to sex, the detection rates were 10.3 % in male and 6.1 % in female (p<0.05), but by age groups, increases of the rates were observed as increase in age (p<0.05). Out of a total of 380 fresh-water fishes of 32 different species, 93 fishes (25 %) of 12 species were found positive with Clonorchis metacercariae, and there were differences in infection rates of the metacercariae among the fishes in 3 parts of the river; 11 % in upper, 35 % in middle, and 34 % in lower reaches respectively. The metacercarial detection rates from various fishes were 97 % in Pseudorasbora parva, Cultriculus eigenmanni (85 %), Gnathopogon strigatus (67 %), Microphysogobio yaluensis (50 %), Gnathopogon coreanus (47 %), Pungtungia herzi(44 %), Abbottina rivularis (40 %), Moroco oxycephalus (33 %), Coreoleuciscus splendidus (32 %), Gnathopogon majimae (26 %), Rhodeus ocellatus (7 %), and Aphyocypris chinensis (3 %) respectively. Although very few P. manchouricus were collected at upper reach, 12 snails (0.7 %) among a total of 1,713 were found infected with Clonorchis cercariae. Also the cercariae of Echinochasmus japonicus (7.99 %), Lexogenes liberum (0.99 %), Cyathocotyle orientalis (0.75 %), Exorchis oviformis (0.23 %) and Asymphylodora japonica (0.05 %) were detected from the snails. PMID- 12902647 TI - [Epidemiological Studies On Clonorchis Sinensis Infection Along The Nam-River In Gyeongnam Province, Korea] AB - An epidemiological study on Clonorchis sinensis infection along the Nam-river (total length; 186km) flowing in Gyeongsang-nam-do, southern part of Korea, was carried out. Formalin-ether concentration technique and Stoll's egg counting method were employed to figure out the prevalence of C. sinensis infection. For the detection of cercariae from Parafossarulus sp., the snail host of C. sinensis, each snail was placed in aerated tap water, and examined for expelled cercariae. For observing the metacercariae the fresh water fish favorably eaten in raw conditions were prepared by means of pressing the muscles between tow slide glasses and/or by digesting them with artificial gastric juice. The fresh water fish were fed to the rabbits to get the worms and to identify the morphology of adult C. sinensis. The results are summarized as follows: 1. Overall prevalence of C. sinensis infection was 38.7% form 5,291 examinees; 44.1% (1,408 out of 3,196) in male and 30.4% (637 out of 2,095) in female. 2. The prevalence rates were 42.0% at the upper stream, 41.2% around the vicinity of Jinyang-lake, 34.2% at Jinju city, 34.2% at middle stream and 40.3% at down stream regions, respectively. 3. By age, the highest positive rate (53.4~54.3%) was observed in 30 to 59 years of age. In this age group, the rate in males was 59.7~62.2%, and in females 42.2~44.4%. In the age group of less than 19years it was 7.5~20.9%. 4. By social strata, the positive rate was 16.5% in the primary school children, 22.6% in school students, 46.2% in teachers and local officers and 49.6% in the general inhabitants. 5. The quantitative examinations with the stool collected from clonorchiasis cases revealed that the light infection (less than 4,000/EPG) was 53.6%, moderate infection (4,001~10,000/EPG) 30.3% and heavy infection (more than 10,001/EPG) 16.1, respectively. More than half of total cases examined were light infection, and 73.2% of female examinees were lightly infected with this fluke. 6. The average value of EPG was 4,963 (male, 6,057; female, 2,557 and the highest value was obtained from the age group of 30 to 59 years (5,240~6,454). 7. The prevalence of Metagonimus yokogawai infection in humans was 5.5%, and 89.8% of metagonimiasis cases were double-infected with C. sinensis. The highest prevalence rate was observed in JInju city(11.2%). 8. Total of 5,005 Parafossarulus sp., the snail intermediate host of C. sinensis, were examined for the detection of cercariae. The cercarial expulsion rate was 0.34%, and the snails collected in Jin-yang-lake side and in the down stream expelled mainly the cercariae of C. sinensis. 9. About 788 cercariae/day (range: 127~1,503) were expelled daily from a snail naturally infected with C. sinensis. The snails which released more than 1,000 cercariae/day were 30.8% out of total collected. A snail uniquely released 5,840 cercariae/day in this study. 10. The other trematode cercariae besides C. sinensis were also detected, and the rates out of total snails were the cercariae of Loxogenes liberum 6.71%, Cyathocotyle orientalis 0.26%, Notocotylus attenuatus 2.52% and Mucobucaris 0.54%. 11. Ten out of 18 species of fresh water fish caught along the river harbored the vetacercariae of C. sinensis. The highest rate of metacercarial infection in fish wa detected in Pseudorabora parva(85.9%). The fish mainly eaten by the inhabitants along the Nam-river, and the metacercarial infection rates were: Zacco platypus 8.0%, Hemibarbus sp. 18.2~26.7%, Gnathopogon sp. 37.5%, Ischikauia steenackeri 42.9% and Pseudogobio esocinus 16.7%. 12. Out of 36 P. parva, the number of metacercariae were about 109 (range; 18~446) per fish and 27 per gram of flesh. The fish caught in spring harbored the highest number of metacercariae. As indecated above, the prevalence of C. sinensis infection in the inhabitants resding around the Nam-river was relatively high. The farther toward the upper stream areas, the higher was the positive rate of C. sinensis infection, and most of clonorchiasis cases were lightly infected. The snail hosts of C. sinensis distributed all around the Nam-river. Several species of freshwater fish were infected with the metacercariae of C. sinensis and the infection rates were relatively high. To prevent C. sinensis infection in the endemic areas, the effective health education system is suggested as a control measure, althought mass treatment is also expected to be useful, using chemotherapeutic agents such as "praziquantel", a recently developed anthelminthic for C. sinensis infection. PMID- 12902648 TI - [Survey On Encysted Cercariae Of Trematodes From Fresh-Water Fishes In Mangyeong Riverside Area] AB - In order to clarify infestation pattern for the encysted larvae of digenetic trematodes from fresh-water fishes, this survey was carried out from March to September, 1983. A total of 380 fishes of 32 species were collected with netting at the three reaches, upper, middle and lower in Mangyeong riverside area. After the fishes were dissected into small scraps, they were pressed under cover glass and examined for the presence of those of digenetic trematodes with a microscope. The results obtained were as follows: Out of a total of 380 fishes inspected, 320 fishes (84 %) from 31 species were found positive with digenetic trematode metacercariae; more than 10 species of the metacercariae were detected in Pseudorasbora parva; Gnathopogon majimae, Microphysogobio yaluensis, Cultriculus eigenmanni and Gnathopogon coreanus (more than 8 species); Aphyocypris chinensis(8 species) and etc. respectively. Clonorchis sinensis metacercariae were found positive from 93 fishes (25 %) from 12 species detection rates in other species of digenetic trematode metacercariae from various fishes were; Exorchis oviformis, 261 fishes (57 %) from 28 species; Cyathocotyle orientalis, 47 fishes (12 %) from 12 species; Metorchis orientalis, 21 fishes (6 %) from 12 species; Metagonimus yokogawai, 164 fishes (43 %) from 26 species; Pseudexorchis major, 71 fishes (19 %) from 18 species; Metacercaria hasegawai, 77 fishes (20 %) from 25 species; Centrocestus armatus, 24 fishes (6 %) from 7 species; Echinochasmus japonicus, 2 fishes (0.5 %) from 2 species, and unidentified species, 34 fishes (9 %) from 15 species respectively. The sums of average number of the encysted larvae of all species found in fish body/gram showed 83 in P. parva, Cobitis taenia (74.2), A. chinensis (28.5), Pseudoperilampus uyekii (26.6), G. majimae (19.6) and etc. respectively and the average peak number of each metacercaria in fish body/gram showed 21.7 C. sinensis, 24 E. oviformis, 15.3 M. orientalis and 6.1 E. japonicus in P. parva; 42.7 C. orientalis and 25.1 M. yokogawai in C. taenia; 8.3 C. armatus and 8.3 M. hasegawai in P. uyekii; 6.3 P. major in Carassius carassius, and 2.9 unidentified species in G. majimae respectively. PMID- 12902649 TI - [Distribution Of Medically Important Freshwater Snails And Larval Trematodes From Parafossarulus Manchouricus And Semisulcospira Libertina Around The Jinyang Lake In Kyongsang-Nam-Do, Korea] AB - The Jinyang Lake is dammed up Nam River, and surrounded by Jinju city and four counties; Jinyang, Sanchong, Hadong, and Sachon in Kyongsang-Nam-Do, Korea. The area around this man-made lake have been known as an endemic focus of clonorchiasis in Korea. The present study was first aimed to know the distribution of freshwater mollusks including medically important snails, and larval trematodes shed from Parafossarulus manchouricus and Semisulcospira libertina. In addtion to above studies, water analyses in each snail habitat were carried out in order to figure out a part of their environmental factors. This malaco-ecological survey was done at the six areas around upper, middle and lower parts of the lake for 4 months, August-November, 1983. Total nine species of freshwater mollusks were collected throughout the study: 4 species of gastropods; Semisulcospira libertina, Cipangopaludina chinensis, Parafossarulus manchouricus and Radix auricularia, and 5 species of bivalves; Unio douglasiae, Anodonta woodiana, Lamprotula gottschei, Corbicula fluminea and Limnoperma lucustris. Out of nine species of freshwater mollusks, three species of gastropods; S. libertina, P. manchouricus and R. auricularia were medically important in terms of the transmission of digenetic trematodes to humans. P. manchouricus and R. auricularia were mainly collected from the shallow ponds and the irrigation channels with the muddy basin, but S. libertina and the bivalves were only collected from the stream of Nam river where the gravels and rocks were dominant. The levels of dissolved oxygen(D.O.) and biochemical oxygen demand (B.O.D.(5)) of the water specimens sampled from the study areas ranged from 6.0 to 9.6 ppm and from 0.4 to 1.6 ppm respectively. As a result, it is considered that water system around the Jinyang Lake might be relatively clean without any heavy pollution of aquatic microorganisms and organic materials during the period of this study. On the other hand, eight metalic constituents from the water samples were also assayed, and all metalic ions detected were remarkably low below the legal criteria. However, calcium ion in the water samples from the habitats of P. manchouricus was considerably higher than others. Infection rates of digenetic trematodes in the snails were 6.9 % in P. manchouricus and 4.8 poercent in S. libertina, respectively. P. manchouricus snails harboring with the cercariae of Clonorchis sinensis were only 0.14 % among the snails examined and other trematode cercariae except cercaria of C. sinensis were; furcocercus cercariae, cercaria of Loxogenes liberum type I and II. S. libertina snails parasitized with the cercariae of Metagonimus yokogawai were 1.5 % out of the snails examined and no cercaria of Paragonimus westermani was found in S. libertina snails in the present study. Digenetic trematode cercariae other then M. yokogawai in S. libertina snails were: Cercaria yoshidae (B type), Cercaria cristata, Cercaria innominatum, Cercaria of Centrocestus formosanus and Cercaria nipponensis. PMID- 12902650 TI - Cestode Fauna Of Hill-Stream Fishes In Garhwal Himalayas, India. AB - Ptychobothrium nayarensis n. sp. has been described from two hill-stream fishes, Barilius bola(Ham.) and Schizothorax richardsonii(Gray) at 325~750 mASL. The new species has been compared with close species. Taxometric evaluation of the new species has been done on available data. PMID- 12902651 TI - A Study On The Structure Of Clonorchis Sinensis, A Liver Fluke: IV. Probable Functions Of The Laurer's Canal. AB - A study on the function of the Laurer's canal of Clonorchis sinensis was conducted with help of the light microscope, the transmission electron microscope, and the scanning electron microscope. Some selected sexual organs concerning with the passages of the spermatozoa and the eggs were obseved in detail. The conclusion of this study is that the Laurer's canal may be the copulatory organ of the female reproductive system as Miyazaki et al. suggested in case of lung flukes. PMID- 12902652 TI - Studies On Intestinal Trematodes In Korea: VIII. A Human Case Of Echinostoma Hortense Infection. AB - Echinostomatid eggs were found from a 21-year old man, whole native village is Munkyong-gun, Kyongsangnam-do during fecal examination in October 1983. The eggs were detected again one week later concommitantly with the eggs of Clonorchis sinensis. He had no subjective symptoms related with these fluke infections. He was treated with praziquantel(Distocide(R)) 20 mg/kg in single dose and purgated with magnesium salt. One echinostomatid fluke and 4 adults of C. sinensis were collected from the diarrheal stools. After morphological observation, the echinostomatid fluke was identified as Echinostoma hortense Asada, 1926. This is the first record on human E. hortense infection in Korea. PMID- 12902653 TI - Studies On Intestinal Trematodes In Korea: IX. Recovery Rate And Development Of Fibricola Seoulensis In Experimental Animals. AB - An experimental study was carried out to observe the susceptibility of several kinds of laboratory animals to Fibricola seoulensis infection, a diplostomatid fluke of mammals. The metacercariae were obtained from the viscera of the snakes, Natrix tigrina lateralis and 50-2,000 in number each was artificially fed to a total of 127 animals; albino rats, mice, dogs, cats, rabbits and chickens. After 3 days to 8 weeks the animals were sacrificed and the recovery rate of worms as well as their maturity was observed. The results are as follows: The overall wom recovery rates throughout the experimental period was highest in albino rats(40.0 %) followed by mice(33.9%), cats(20.9 %), dogs(11.4 %), rabbits(0.05 %) and chickens(0 %). However, the recovery rates in the same host decreased as infection progressed longer and variable by the amount of metacercariae given. From albino rats and mice, the highest recovery rates were obtained in 1,000 and 200 metacercariae infection groups repectively, and it is considerd that such amount should be the optimum dose for experimental infection of these animals. The main location of F. seoulensis in experimental animals was small intestine especially the duodenum. The maturity index (No. mature worms/No. examined) was 100% in albino rats and mice, while only 22.7% or 0% in dogs or cats respectively. From the results, it is concluded that albino rats and mice are the most susceptible hosts for F. seoulensis infection among six kinds of laboratory animals examined. PMID- 12902654 TI - [Changes In The Pathogenicity Of Naegleria Fowleri By Several Brain Passage In Mice] AB - The pathogenicity of free-living amoeba, Naegleria fowleri, is influenced according to the strain, cultural condition and host (Culbertson et al., 1968; Carter, 1970; Wong et al., 1975). Phillips (1973) demonstrated that Entamoeba histolytica became avirulent after more than 2 year maintenance in axenic culture in vitro. This study was carried out to compare the difference in pathogenicity between two strains of N. fowleri, one of a prolonged maintenance in axenic medium and the other one obtained by serial brain passage in mice. The 0 strain was that N. fowleri had cultivated axenically more than 7 years in CGVS medium. The 2-1 strain was obtained from the brain of mouse inoculated intranasally with a strain, which was from the mouse brain infected with 0 strain, and cultured for 15 weeks until the beginning of this experiment. White male mice weighing 18-22 g were used. Mice were anesthetized by an intraperitoneal injection of about 1 mg secobarbital, and inoculated intranasally with 10 x 10(4) live N. fowleri trophozoites in a 5 microliter cell suspension. Sluggish behaviour, nervousness, rotation and leg paralysis were developed earlier and more frequently in the 2-1 experimental group than the control 0 group. Pathological changes such as inflammatory and necrotic lesion were observed in the olfactory and anterior portion of brain, and these changes were more extensive in the 2-1 group. The edematous and inflammatory changes in lung were demonstrated in mice died after 13th day post-inoculation. The experimental mice of 2-1 group began to die suddenly from 7th day post-inoculation, and the survival time in 2-1 group mice was shorter than 0 group mice. The typical primary amoebic meningoencephalitis was developed in the mice inoculated intranasally with N. fowleri. The prolonged maintenance of N. fowleri amoebae in axenic CGVS medium was observed to have lost their original pathogenicity for mice, but their pathogenicity was restored by serial brain passage in mice. PMID- 12902655 TI - Praziquantel(Distocide(R)) In Treatment Of Clonorchis Sinensis Infection. AB - Praziquantel(Distocide), the Korean product, was tested for its safety and efficacy in treatment of Clonorchis sinensis infection during the period from April to September, 1983 in Korea. A total of 55 egg positive cases were selected and treated with the regimen of 25 mg/kg t.i.d. for 1 day (total 75 mg/kg). The follow-up stool examination was done in 47 cases by cellophane think smear and Stoll's egg counting techniques. The 8 uncured cases were treated again with the same regimen. The laboratory tests for blood picture and liver function were done in 27 cases and compared before and after the treatment. The results obtained are as follows: After single course treatment, the cure and egg reduction rates were 83.0% and 99.1% respectively. With the second treatment, excellent results of 100% in both rates were obtained. Several kinds of side effects such as dizziness, headache, etc. were complained by 29 cases (61.7%), however, those were so mild and transient that no special treatment was necessary. No significant chang in laboratory findings was recognizable before and after the treatment. From the above results, it is concluded that Distocide is as effective and safe as Biltricide(R) and highly recommendable in treatment of C. sinensis infection. PMID- 12902656 TI - [An Epidemiological Study Of Human Paragonimiasis By Means Of Micro-ELISA] AB - As epidemiological parameters of human paragonimiasis, the positive rates of intradermal test and the sputum/stool examination have long been employed in population surveys. However, both the specificity of the intradermal test and the sensitivity of sputum/stool examination have been gradually declined as the endemicity was lowered; thus the gap between above two parameters widened. In such context, the development of a new epidemiological parameter or tool which makes it possible to accurately discriminate the active paragonimiasis cases was necessary. In the present study, the detection rate of Paragonimus-specific IgG antibody by micro-ELISA was evaluated as an indicator of epidemiologic status of human paragonimiasis in a population. A total of 4,285 students and inhabitants living in Bukpyeong Myeon and Bukil Myeon, Haenam Gun, Jeonlanam Do was surveyed in October 1983 by intradermal test first. Out of them, 244 case (5.7 %) were found positively reacted to VBS antigen of P. wetermani. Out of 168 positive reactors, 7 cases (4.2 %) were egg positive either by two times of sputum examination or by one stool examination. That indicated that only 0.16 % of total surveyed were confirmed as active paragonimiasis by egg detection. When sera collected from 239 positive reactors of intradermal test were tested by micro ELISA for their specific IgG antibody, 40 cases (16.7 %) were found to be positive. All of 7 eggs positive cases were again positive for specific IgG antibody. Among remaining 232 intradermal test positive cases, 33 cases were positive for IgG antibody. In contrast to those, none of 42 positive reactors to intradermal test for Clonorchis and of 128 intradermal test negative cases showed positive for Paragonimus- specific IgG antibody. The rate of specific IgG antibody as detected by micro-ELISA appeared to be increased with the wheal size of the intradermal test. When the wheal size was over 13 mm in diameter, about 50 % of them were positive for specific IgG antibody. Thirty-one specific antibody positive cases were clinically evaluated by laboratory examinations (repeated sputum examination, peripheral eosinophil count and chest roentgenogram) and by history taking. Out of them 24 cases were associated with one or more positive laboratory findings; thus considered as active paragonimiasis cases. Out of 7 lab. finding-free cases 3 revealed past history of typical paragonimiasis symptoms, suggesting that they were in chronic or in convalescent stages. The remaining 4 cases were considered as either mild or ectopic infection cases; the possibility of cross-reaction with other helminthiases could not be ruled out. From the above results, it was inferred that the detection of Paragonimus specific IgG antibogy by micro-ELISA was very much helpful in detecting the active cases as well as in proper evaluation of the endemicity of human paragonimiasis in a population. The convenience of mass handling of sera in micro ELISA was considered another superiority as an epidemiologic tool. PMID- 12902657 TI - [On The Applicability Of Partially Purified Antigenic Preparations Of Paragonimus Westermani] AB - In order to obtain more specific antigenic preparation for the diagnosis of human paragonimasis, crude saline extract of whole worm (=PwWWE), secretory-excretory components (PwSEC) and secretion-excretion-free somatic extract (PwSM) of 12 week old Paragonimus westermani were filtrated through Sephadex G-200 gel column. The adult Paragonimus worms were obtained from experimentally infected dogs. A total of 11 antigenic solutions was lyophilized or diluted to adjust protein content of 1 mg/ml. To evaluate the antigenicity of crude antigens and fractions, micro ELISA was done with the sera from P. westermani infected cases, C. sinensis infected cases and non-infected control cases to detect Paragonimus specific IgG antibody. The results were as follows: When the PwWWE was filtrated through Sephadex G-200 gel, it was separated into three fractions; PwWWE Fr. 1, PwWWE Fr. 2 and PwWWE Fr. 3. The percentage of protein content was 28.0 %, 21.6 % and 50.4 % respectively. The PwSM was also separated into three fractions; PwSM Fr. 1, PwSM Fr. 2, PwSM Fr. 3 and their percentage of protein content was 41.3 %, 38.6 % and 20.1 %. However, the PwSEC showed different fractionation pattern; i.e. fraction 1 (=PwSEC Fr. 1) and 3 (PwSEC Fr. 3) without fraction 2. The percentage of protein content was 14.0 % in PwSEC Fr. 1 and 86.0 % in PwSEC Fr. 3. When the antigenicity of each Paragonimus crude antigen and fractionated antigen was evaluated for specific IgG antibody by micro-ELISA in 10 human paragonimiasis sera, PwSEC Fr. 1 was the most potent antigen showing the mean absorbance 1.98. The PwWWE Fr. 1, PwSEC, PwWWE were next to that; their mean absorbance were 1.72, 1.38 and 0.83, respectively. The antigenicity of fractions 2 and 3 was much weaker in binding specific IgG antibody. When the antigens were reacted in micro ELISA with 10 human clonorchiasis sera, most antigens showed weak reactivity. Each fraction 1 of crude antigens reacted higher than other fractions or crude antigens; the mean absorbance was 0.17 in fraction 1, but in others the absorbances were about 0.06. With non-infected control sera, the result of micro ELISA revealed almost same pattern with those of the clonorchiasis sera. From the above results, it became apparent that PwWWE Fr. 1, especially PwSEC Fr. 1 was the most potent antigen reacted with Paragonimus specifc IgG antibody. PMID- 12902658 TI - [Comparison Of Tia With ELISA For Circulating Antibody Detection In Clonorchiasis] AB - A comparison was made of a new serological method, thin layer immunoassay (TIA), and an established method, enzyme-linked immunosorbent assay (ELISA), in the detection and quantification of antibodies in clonorchiasis. Saline extract of lyophilized Clonorchis sinensis adult worm was used as antigen, and TIA by the method of Elwing et al. (1976) and ELISA by Voller et al. (1974) were performed. Using sera from known clonorchiasis cases, 100 % of the sera tested were positive by TIA and 88.3 % by ELISA. TIA produced false positive results in 14 out of 36 cases, which were 10 amoebiasis cases, 16 paragonimiasis cases and 10 healthy controls. ELISA, however, produced a small number of false positives, 7 out of 55 cases. There was correlation between immunoglobulin G level in sera and ELISA value (correlation coefficient, 0.69), whereas no correlation between immunoglobulin G level and TIA result. The performance of TIA and ELISA was not correlated in the results using homologous antigen. PMID- 12902659 TI - [Comparison Of Agar-Gel Diffusion Tests, Counterimmunoelectrophoresis And Enzyme Linked Immunosorbent Assay In The Sera Of Skin Test Positives For Paragonimiasis] AB - Agar-gel diffusion test (AGD), counterimmunoelectrophoresis (CIEP) and enzyme linked immunosorbent assay(ELISA) were examined with the sera of skin test positives for paragonimiasis. The crude antigen(Paragonimus whole worm extracts: protein concentration, 7.56mg/ml) and human sera were used in AGD and CIEP. And in ELISA test, diluted antigen with 1:40,000 of crude antigen and diluted sera with 1:100, 1:200 were used in the test. The positive identical ratio between AGD and CIEP reactions is 98 % and negative identical ratio is 100 %. One or three precipitin bands are observed in AGD. One to seven precipitin bands are also revealed in CIEP. Especially, deeply stained bands are observed in CIEP than those of AGD. The positive identical ratios between AGD and ELISA tests are 96 % in 1:100 diluted sera, and 94 % in 1:200 diluted sera. But the negative identical ratios between AGD and ELISA tests are 97 % and 99 % respectively in 1:100 and 1:200 diluted sera. The positive identical ratios between CIEP and ELISA tests are 98 % and 96 % respectively in 1:100 and 1:200 diluted sera, but also 97 % and 99 % in 1:100 and 1:200. Control sera, such as clonorchiasis, amoebiasis and toxoplasmosis, revealed all negatives with Paragonimus antigen in AGD, CIEP and ELISA tests. By above results, ELISA was most sensitive, next CIEP and AGD. But AGD test appears to be more useful when used to crude antigen without cross reaction with other parasitic infections. CIEP test is basically equal in terms of precipitin reaction, but CIEP is able to be detected more sensitively and rapidly though less simple in handiwork than AGD. Consequently, three methods for immunological tests of paragonimiasis have good correlations with one another. Also, each of these has both merits and demerits in immunological test for paragonimiasis. But the ELISA test was proved to be the most sensitive and convenient tool for mass screening test, especially in case of using purified antigen. PMID- 12902660 TI - [Toxoplasma Antibodies By Indirect Latex Agglutination Tests In National Seoul Mental Hospital Patients] AB - A total of 573 patients hospitalized in National Seoul Mental Hospital and 76 of healthy persons as control were examined by indirect latex agglutination test in order to evaluate Toxoplasma antibody titers in mental patients. Throughout this survey, 1:32 or more titers of diluted sera were regarded as positive. The 573 samples of test sera showed negative in 386 cases (67.4 %), 1:2 in 93 cases (16.2 %), 1:4 in 57 cases (9.9 %), 1:8 in 14 cases (2.4 %), 1:16 in 12 cases (2.1 %), 1:32 in 5 cases (0.9 %), 1:64 in 1 case (0.2 %), 1:128 in 3 cases (0.5 %) and 1:256 in 2 cases (0.3 %) respectively. Among total 573 mental patients, 11 cases (19 %) showed positive, and they were 9 cases (1.8 %) of schizophrenia and 2 cases (7.4 %) of manic depression. One case (1.3 %) out of 76 control sera showed positive result. PMID- 12902661 TI - [Detection Of Paragonimus-Specific Igg Antibody In Csf And Pleural Effusion By Micro-ELISA] PMID- 12902662 TI - [Thin Layer Immunoassay (Tia) For Circulating Antibody Detection In Clonorchiasis] AB - Thin layer immunoassay was carried out to demonstrate antibodies against Clonorchis sinensis in sera from clonorchiasis patients. Saline extract of adult worm was used as antigen. TIA technique was performed as described earlier by Elwing et al. (1976), but agarose was used instead of agar. The antibody titres of sera in 60 clonorchiasis cases were higher than that of 10 healthy and 10 amoebiasis cases, but not different comparing with that of 10 paragonimiasis cases. Antibody titres in clonorchiasis gave no differences according to the age, sex, EPG in feces, eosinophilia degree of blood, level of alkaline phosphatase and transaminase (SGOT, SGPT) in sera. It is suggested that, after evaluation, the TIA might supplement or be used as an alternative to other immunodiagnostic tests already in use for the diagnosis of clonorchiasis. PMID- 12902663 TI - [Infestation Of Larval Trematodes From Fresh-Water Fish And Brackish-Water Fish In River Taechong, Kyungpook Province, Korea] AB - A study of infestation patterns for larval trematodes from fresh-water fish and brackish-water fish was carried out during the period from June to October in 1982. They were collected by netting, fishing with rod and line, and using the bait of crushed oil cake in a transparent plastic bowls at the three localities, Daebon-dong, Kugil-dong, and Hoam-dong in the river Taechong, Kyungpook Province, Korea. Five species of fresh-water fish, the crussian carp, Carassius carassius Linnaeus, the bullhead, Coreobagrus brevicorpus Mori, the fat minnow, Moroco oxycephalus (Bleeker), the flat bitterling, Paracheilognathus rhombea(Temminck et Schlegel), and the catfish, Parasilurus asotus (Linnaeus), and two kinds of brackish-water fish, the grey mullet, Mugil cephalus (Linnaeus) and the sweetfish, Plecoglossus altivelis (Temminck et Schlegel) were examined. Four species of the metacercariae, Exorchis oviformis, Metacercari hasegawai, Metorchis orientalis and undetermined larvae were found. Of these, the encysted larvae of the human intestinal fluke, Metagonimus yokogawai, were found from three kinds of fresh-water fish, the crussian carp, the bullhead and the fat minnow and one species of brackish-water fish, the sweetfish and the infestation rates for the larvae varied appreciably from fish to fish. The intensity of infestation with the Metagonimus larvae in the three species of fresh-water fish as low, and the average number of the larvae per gram of flesh varied from 0.5 to 0.2. By comparison, the brackish-water fish was more heavily infested and the mean number of cysts per gram of flesh in Plecoglossus altivelis was 109.2. It was found that infection with Metagonimus yokogawai is caused by consuming raw fresh-water fish and brackish-water fish caught in the river Taechong, Kyungpook Province, Korea. PMID- 12902664 TI - [Experimental Infection Of Clonorchis Sinensis To Cyprinus Carpio Nudus] AB - As a part of observation on the suitability as a second intermediate host of Clonorchis sinensis, various infection experiments were tried to Cyprinus carpio nudus. The results obtained were summarized as follows: 1. The cercariae which attempted to contact with Cyprinus carpio nudus in the water were observed under the stereomicroscope. After contact, cercariae began to separate its tails from the bodies at 4 minutes increasingly, and then the number ranged to 80 percent at 13 minutes after the encounter. But very few cercariae could actually invade into the epidermis of the fish. 2. When Cyprinus carpio nudus were exposed to a number of cercariae in the beaker, only a few cercariae could invade through the epidermis. Most of the invaded cercariae were killed before forming the cyst. Also rare encysted cercariae were found to be dead within 48 hours. 3. Cyprinus carpio nudus were reared with Parafossarulus manchouricus shedding numerous cercariae in the indoor aquarium. When observed after 12 days, to determine the susceptibility, no metacercaria was found infected. 4. Cyprinus carpio nudus were reared with Parafossarulus manchouricus shedding numerous cercariae in Kimhae lake for 2 months in summer of 1982, in order to check the natural infection. No metacercaria of Clonorchis sinensis was found from the fish. 5. In the cases of control fish, Pseudorasbora parva, numerous cercariae were inspected on the course of invasion through epidermis of the fish under the stereomicroscope. And many metacercariae were also found from the fish whenever they were kept in beaker, indoor aquarium, or the lake for natural infection. The results suggested that Cyprinus carpio nudus might not be proper intermediate host of Clonorchis sinensis. PMID- 12902665 TI - [The Wormicidal Substance Of Fresh Water Fishes On Clonorchis Sinensis: Iv. Preliminary Research On The Wormicidal Substance From Mucus Of Cyprinus Carpio Nudus] AB - As a series of studies on the suitability as a second intermediate host of Clonorchis sinensis, the wormicidal effects of mucus from epidermis of Cyprinus carpio nudus were observed. The results obtained were summarized as follows: On the silica gel thin layer chromatography with the greenish yellow supernatant in acetone:benzene/10 : 90 as carrier, seven spots were observed in iodine chamber. The spot of Rf. 0.225 value among them disclosed the strongest wormicidal effect on the cercaria, the excysted metacercaria, and the adult of C. sinensis. Meanwhile, no seasonal variations were found in Rf. values and effect of the wormicidal fractions. The wormicidal materials in epidermal mucus of C. carpio nudus and Carassius carassius were compared in their effect, and the former was more effective killing substance than the latter. It seemed that C. carpio nudus could not be proper intermediate host of C. sinensis according to results of the present study, in addition to our previous observation results. PMID- 12902666 TI - Evaluation Of Specific Ige Antibody In Clonorchis Sinensis Infection. AB - Serum IgE level and Clonorchis specific IgE in individuals with Clonorchis sinensis were determined by radioimmunosorbent(RIST) and radioallergosorbent technique(RAST) respectively. Highly significant elevations of serum IgE (P<0.001) and specific IgE antibodies (P<0.01) were observed in area from individuals with clonorchiasis. The mean values of serum IgE in individuals with clonorchiasis and healthy individuals were 2,372 IU/ml and 364 IU/ml respectively and specific IgE antibodies of both groups were 52.0 and 4.4%. A close correlation(r=0.9451) between serum IgE level and specific IgE antibodies were observed and correlation (r=0.6056) between serum IgE and EPG and between specific IgE and EPG(r=0.5693) were also observed. PMID- 12902667 TI - Lipofuscin Pigment In Adult Clonorchis Sinensis. AB - Adult Clonorchis sinensis from infected human cases may have black pigment in their parenchyme. In present paper, the nature of the pigment was investigated. The pigment distributed in cytoplasm of parenchymal cells of the worms. Histochemically the pigment was positive by periodic acid-Schiff, Ziehl-Neelsen, and lipofuscin stainings. The pigment particle contained many, various sized membrane structure as well as electron dense granules when observed by electron microscopy. Staining characteristics of the black pigment, together with electron microscopic findings, strongly indicated that the pigment was lipofuscin. PMID- 12902668 TI - [Aspartate And Alanine Aminotransferase In Fasciola Hepatica] AB - The activity and distribution of aspartate aminotransferase (EC 2.6.1.1) and alanine aminotransferase (EC 2.6.1.2) in adult Fasciola hepatica have been studied. Fasciola hepatica was fractionated by differential centrifugation into nuclear, mitochondrial and cytosolic fractions. The activity of GOT and GPT was measured by the method of Reitman and Frankel. Isozyme patterns of those enzyme were also examined by DEAE-cellulose column chromatography. The results obtained were as follows: 1. The activity of aspartate and alanine aminotransferase was about 0.55 unit and 0.92 unit per 1 g of Fasciola hepatica, respectively. 2. The activity of those enzymes was relatively low compared with those in mammalian tissues. 3. The distribution of aspartate aminotransferase in the subcellular organelles showed that 71 % of the activity was in cytosolic, 24 % in mitochondrial and 5 % was in nuclear fraction. 4. About 22 % of the total alanine aminotransferase activity was found in the mitochondrial fraction, about 66 percent in the cytosolic fraction. 5. Aspartate aminotransferase from cytosolic fraction was separated into two types of isozymes, whereas alanine aminotransferase from cytosolic fraction gave only one active peak on DEAE cellulose column chromatography. PMID- 12902669 TI - [Purification And Properties Of Branched Chain Amino Acid Aminotransferase From Fasciola Hepatica] AB - The distribution and properties of branched chain amino acid aminotransferase(EC 2.6.1.42) was investigated in adult Fasciola hepatica. Fasciola hepatica was fractionated by differential centrifugation into nuclear, mitochondrial and cytosolic fractions. The activity of branched chain amino acid aminotransferase was measured by the method of Ichihara and Koyama (1966). Isozyme patterns of this enzyme was also examined by DEAE-cellulose column chromatography. The results obtained were as follows: 1. The activity in homogenate was found to be 12.69 units/g wet tissue. The activity of this enzyme was relatively high compared with those in rat tissues. 2. The distribution of branched chain amino acid aminotransferase in the subcellular organelles showed that 87.8 % of the activity was in cytosolic, 10.9 % in mitochondrial and 1.3 % was in nuclear fraction. 3. Cytosolic fraction of Fasciola hepatica contained Enzyme I, but not Enzyme II and III, of branched chain amino acid aminotransferase. Enzyme I was eluted by 50 mM phosphate buffer from DEAE-cellulose column and catalyzed the transamination of all three branched chain amino acids. 4. The Enzyme I was purified about 22-folds increase in specific activity after chromatography on DEAE-cellulose. 5. The best substrate among three amino acids (leucine, isoleucine and valine) was L-isoleucine. 6. The optimal temperature of Enzyme I was 45 degrees C and the optimal pH was 8.2. 7. The Km value for leucine of Enzyme I was 4.17 mM. 8. The Km values for alpha-ketoglutarate and pyridoxal phosphate of Enzyme I were 0.41 mM and 4.76 x 10(-3) mM, respectively. PMID- 12902671 TI - [Studies On The Bladder Worm, Cysticercus Cellulosae: The Ultrastructure Of C. Cellulosae] AB - An electron microscopic study was performed to know the basic tegumental structure of Cysticercus cellulosae. The scolex and bladder portions of cysticerci (human and porcine strains) were prepared for transmission and scanning electron microscopy by conventional procedures. In general, the tegument of C. cellulosae showed the basic ultrastructure of cestode tegument on electron micrographs. The teguments of both scolex and bladder portions consisted of such components i.e., an outer vesicular layer with numerous microtriches and inner fibrous layer. Below the fibrous layer, there were layers of muscle bundles and tegumental cells. The microtriches which covered the surface of cysticercus revealed two distinctly different shapes. The characteristic bladder-like, elongated pyramid shaped 'tetrahedral form' was observed on the surface of the scolex portion, whereas the elongated cylindrical 'filamentous form' was distributed on the surface of bladder portion. In spite of the difference of isolated host and location, the cysticerci showed the same result. But dimensional variations of the tegument according to topography of the worm were observed. The possibility of application in making differential diagnosis from other larval cestodes and possible functions of this larval tegument were discussed. PMID- 12902670 TI - A Study On Intestinal Lesions Of Experimentally Reinfected Dogs With Metagonimus Yokogawai. AB - The intestinal lesions were studied in dog metagonimiasis by reinfection. The metacercariae of M. yokogawai were collected from naturally infected sweetfish. A total of twenty dogs were divided into three groups; control, primary infection and reinfection groups. The 18 dogs were infected with 10,000 metacercariae; the 5 dogs in primary infection group were killed at the 5 days in 1 week, 4 weeks, 6 weeks after the infection. The remaining 13 dogs were kept for nine weeks, and challenged again with 5,000 metacercaricae. The reinfected dogs were killed at the 1 day, 3 days, 5 days and 1 week, 2 weeks, 4 weeks, 6 weeks, 8 weeks respectively. The chronologic changes of the intestinal lesions in both groups, the worm recovery rate, and distribution patten were examined by gross observation, light and scanning electron microscopies. The results were summarized as follows: 1. Worm recovery rate in primary infection group ranged from 19.8% to 37.8%. The rates were 6.3% to 16.3% in reinfected dogs within 5 dats. And the ratio of young worms collected to old ones was 1:1.83. In primary infection group, the collected worms were distributed in upper portion of the small intestine, while the distribution of worms shifted down to the lower portion of the small intestine in reinfection group. 2. In primary infection group, gross pathologic findings of intestinal mucosa began to appear in the 5th day and the 1st week after infection. The changes were effacement of transverse folds, watery or serous intestinal content, and enlargement of Peyer's patches as well as mesenteric lymph nodes. After the 4 week of infection, these findings returned to normal. In the reinfection group, these findings did occur again, but were lighter in degree when compared with primary infection group in early phase of infection. Again the gross changes returned to normal in two weeks which resolved earlier than in promary infection groups. 3, By histopathologic observations, mucosal changes such as blunted, adhered villi with hyperplasia of crypt epithelium, stromal changes, edema, and inflammatory cell infiltration were observed. These regressed mucosal changes were returned to almost normal, except for inflammatory cell infiltration. The reinfected dogs showed less severe lesions of the small intestine, with shorter period of the changes especially in aspect of villous changes. 4. By the scanning electron microscopy, the histopathological findings of villi lesions were also confirmed. From the above results, it was inferred that the so-called spontaneous and/or self cure phenomena were of negligible importance in metagonimiasis in dogs. In the chronologic observations of pathologic process in intestine, it was also suggested that the host immunity tend to recover the intestinal integrity. And early invasion of young worms to the Lieberkuhn's crypt was considered as the primary cause of lesions of villi. PMID- 12902672 TI - [The Infection Status Of Sparganum And Gnathostoma In Frogs Of Southern Part Of Korea] AB - A survey on the infection status of sparganum and Gnathostoma in frogs was carried out in 11 areas in Gyeongsangnam-do and Cheonlanam-do province, a southern part of Korea. 1. Among 626 frogs examined, 4 percent were infected with sparganum, but none with Gnathostoma. The infection rates of sparganum were variable according to the areas. Those from Kimhae, Sunchun, Jinhae, Sachun, Kosung, Keoje, and Hadong were 16 %, 9.1 %, 9.1 %, 4.1 %, 2 %, 2 %, and 1.5 % respectively, but frogs from Hapchun, KwangJu, Namwon and Sanchung were not found infected. 2. Rana nigromaculata was the only species infected with sparganum. Rana rugosa and Bombina orientalis were not infected. 3. Sparganum was mainly found in the legs and the trunk of frog. About 92.5 % of worms were detected in the femoral intermuscular connective tissue. 4. Number per infected frog was in range of one to five, and 2.8 % of frogs were infected by only one worm. 5. The heavier the frog weight, the higher the infection rate of sparganum. In frogs of 51-60 grams, 28.6 % were infected. PMID- 12902673 TI - An Imported Case Of Kala-Azar In Korea. AB - An imported case of Kala-azar in a 26-year-old Korean man is reported. The diagnosis was made by liver needle biopsy. Amastigotes were seen in Kupffer cells under light microscope, and their characteristic ultrastructural features were recognized under the electron microscope. This case represents an imported disease from Saudi Arabia where the patient spent one year as a construction worker, 8 months before the onset of the disease. This report also signifies the second description of Kala-azar in Korea, and the first case of Kala-azar imported from Saudi Arabia. This patient was successfully treated with sodium antimony gluconate (Pentostam(R)), and follow up liver biopsy showed focal fibrous scar and otherwise normal liver without demonstrable organism. PMID- 12902674 TI - Efficacy Of Mebendazole In Treatment And Control Of Trichuriasis In Korea. AB - Mebendazole was tested for its efficacy in individual treatment and mass control of Trichuris trichiura infection in Korea. The results were as follows: 1. The efficacy of mebendazole in 430 cases 3 weeks after treatment with mebendazole(100 1,200 mg) was not so satisfactory (15.0-64.0 and 24.6-95.2% in cure and egg reduction rates respectively). But it was observed that the higher the total dose of the drug the better the efficacy. 2. The egg positive rate of the inhabitants in mass control programme by 600 mg mebendazole decreased form 40.0 to 5.6 % during one year, May 1982-May 1983, by repeated blanket mass chemotherapy every 3 months (in conventional regimen), while the decrease was less marked or unrecognizable in 3-month (signle or two divided doses), 6-month, 12-month interval and placebo control groups. From the results, it was concluded that, although mebendazole was not highly effective in individual treatment of T. trichiura infection, it was fairly useful in mass cotrol, by repeated mass chemotherapy every 3 months. PMID- 12902675 TI - [A Study On The Intestinal Helminths Of The Patients In A Leprosarium In Korea] AB - A total of 2,026 leprosy patients of the National Sorokdo Hospital was examined their intestinal parasites by cellophane thick smear method in January 1983. The egg positive cases of Taenia spp. were treated with bithionol and the segments of Taenia were collected for species identification. The results were as follows: 1. Total egg positive rate of any kind helminth was 78.2 % and cumulative total was 85.2 %. The egg positive rate for each helminth was as follow; Taenia spp. 3.4 %, Ascaris lumbricoides 4.5 %, Trichuris trichiura 72.l %, Clonorchis sinensis 2.8 % and other 0.05 %. 2. A total of 66 Taenia egg positive cases was treated; out of them proglottids of Taenia were collected from 26 cases. All of the collected worms were identified as T. saginata. The results revealed significantly high egg positive rate of T. trichiura. However, A. lumbricoides was found to be controlled considerably by repeated chemotherapy during past 3 years. If chemotherapeutic agent is replaced with oxantel-pyrantel tablet, better result is expected. No clue was found for prevalence of T. solium from both human and the pig in the island. PMID- 12902677 TI - Description Of Aedes (Aedimorphus) Alboscutellatus Occuring In Korea. AB - Aedes alboscutellatus was recorded first in Korea. This mosquito was collected near Demilitarized Zone of Korea Penninsula from 1979-1980. Its morphology and taxonomical position was described. PMID- 12902676 TI - [Change In Incidence Of Parasite Eggs And Larvae From Vegetable In The Markets Of Taegu] AB - A survey on vegetables, which consisted of lettuce (Lactuca sativa), young radish (Raphanus sapivus), and chinese cabbage (Brassica pekinensis) collected from 6 markets in the Taegu city, was conducted for the discovery of helminth eggs and larvae, from July 1982 to June 1983. The results were compared with the data obtained from vegetables collected at the same markets by Lee (1969) and Choi and Lee (1972). Both sides of vegetable leaves were washed carefully with a hard brush, and the species of parasites and the mean number of parasites per 200 g of vegetable were determined. When vegetable were examined, 4 species of parasite eggs (ascarid, Trichostrongylus, liver fluke, and hookworms) and 2 larvae (filariform and rhabditoid larvae) were found. Of the parasite discovered, ascarid egg was found to be highest (4.2 %), followed by hookworm egg (3.6 %). The mean number of ascarid egg per 200 grams of vegetable was 0.6 in young radishes, and 0.3 in both lettuces and chinese cabbages. Similarly, the number of filariform larva of hookworm was 0.4 in young radishes, 0.3 in lettuces, and 0.l in chinese cabbages, and the number of rhabditoid larva was 0.2 in young radishes, 0.1 in lettuces, and 0.05 in chinese cabbages. In the results obtained by Lee (1969) and Choi and Lee (1972), the mean number of ascarid egg per 200 grams of vegetable was 7.5 in young radishes, 3.l in lettuces, and 0.5 in chinese cabbages. By contrast, in the present study it was 0.6 in young radishes and 0.3 in both lettuces and chinese cabbages. These results suggest that there has been significant reduction in the incidence of parasite egg and larva on vegetable leaves during the past 10 years. PMID- 12902678 TI - [Classification Of Planorbidae Cellected From Geum River Basin] AB - The distribution, external morphology, radula, chromosome numbers of Planorbidae snails were studied. 1. The specimens were collected at four stations in Nonsangun, Kongjugun, and Daedukgun which are located around Geum river. Three genera and three species of Planorbidae, Hippeutis cantori, Segmentina hemisphaerula and Gyraulus convexiusculus, were collected. H. cantori was the most abundant species among the three species. G. convexiusculus was the least abundant one. Each species could be identified on the basis of its external characteristic, since the periphery of each species has a peculiar shape. H. cantori was the largest one among the three species. The radula formula of each species was very similar to other species. The size of radula was proportional to the size of shell. The radula formulae of H. cantori, S. hemisphaerula, and G. convexiusculus were 29:1:29, 23:l:23, and 16:1:16 respectively. The difference of radula formula could be found in the total numbers of laternal and marginal teeth. The haploid chromosome number of H. cantori was eighteen (n=18). S. hemisphaerula and G. convexiusculus were assumed to be same in their chromosome numbers (n=18). PMID- 12902679 TI - Influence Of Transferred Anti-Nematospiroides Dubius Immune Serum And Igg On Immune Reaction In Recipient Mice. AB - The effect of transferred immune serum and IgG on the response of recipient mice to concurrent infections with larvae has been investigated. Following one infection, recipient mice harboured fewer worms than control mice, the worms were stunted and thier fecundities were greatly reduced. Four concurrent infections was found to reduce worm burden and worm fecundity, and induce an increase of serum antibody titers in no-treatment and normal serum given mice. On the other hand the transferred immune serum and IgG appeared to adversely affect the protective immune response in recipient mice. In recipient mice, worm fecundity was consistently increased throughout experiment and harboured more worms after four infections than control mice. Serum anti-N. dubius antibody titers of mice made passively immune by transfer of immune serum and IgG were not significant difference between one infection and four infections. Thus, transferred immune serum and IgG may debilitate host antibody production. PMID- 12902680 TI - Recent Demonstration Of Helminth Eggs And Larvae From Vegetable Cultivating Soil. AB - A survey on soil samples collected from the vegetable gardens and vinyl houses in the vicinity of Taegu was conducted for the discovery of helminth eggs and larvae from October 1981 to September 1982. The samples collected were examined utilizing the "Commonly Employed" and Kamiya's techniques. The details of comparative features of larval nematodes are taken from the handbook of parasitology by Matsubasyhi et al. (1965). The results were compared with the data obtained from soil samples collected at the same area by Lee(1976). Five kinds of helminth eggs, ascarid, hookworm, liver fluke, Trichostrongylidae, and whipworm and undetermined eggs were found. Of these, the incidence of ascarid eggs was found to be the highest, 70.5 per cent, followed by whipworm and hookworm eggs, 9.9 and 9.2 per cent, respectively. The average number of ascarid eggs per 100 grams of soil was higher in the vinyl houses, compared with the number in the vegetable gardens than in the vinyl houses. By contrast, the number of hookworm eggs was relatively high in the vegetable gardens. In the monthly incidences of helminth eggs, ascarid eggs were found every month, continuing fairly high incidences all the year round. However, hookworm eggs was found for 8 months from February and September, with low monthly incidences between 4.0 and 22.2 per cent, and Trichostrongylidae eggs were found for 6 months from April to September, with lower incidences than those of hookworm. Three kinds of larval nematodes, the filariform and rhabditoid larvae of hookworm, Trichostrongylidae, and undetermined larvae were found in the soil of farms, but the incidences of larvae were quite low. The average number of larvae per 100 grams of soil was 0.05. When the number of eggs per 100 grams of the soil in 1976 were compared with 1982, pronounced reduction of the number was encountered in all helminth eggs. However, the incidences and the average number of larvae per 100 grams of the soil were so low that it was impossible to compare the data between 1976 and 1982. PMID- 12902681 TI - Studies On Intestinal Trematodes In Korea: V. A Human Case Infected By Fibricola Seoulensis (Trematoda: Diplostomatidae). AB - A 25-year old Korean male was admitted to Seoul National University Hospital on July 5, 1982 because of fever, epigastric discomfort and diarrhea. Laboratory examination revealed moderate eosinophilia, and large operculated helminth eggs in stool repeatedly. Other laboratory test resulted in normal range and bacteriological cultures were nefative. Under the impression of intestinal fluke infection, bithionol treatment with magnesium purgation was tried. From the diarrheal stool after treatment, a total of 79 small adult trematodes was collected, and identified as Fibricola seoulensis. The patient had eaten raw viscera of 2 snakes 9 days before admission in a rural village in Korea. It was assumed that this is the first record that described human infection by an adult diplostomatid fluke. PMID- 12902682 TI - Studies On Intestinal Trematodes In Korea: VI. On The Metacercaria And The Second Intermediate Host Of Fibricola Seoulensis. AB - This study was carried out to confirm the infection source of the human case of Fibricola seoulensis, and to reveal out a part of its life cycle in Korea. Also the morphological characteristics of the metacercaria were described. The results were summarized as follows: 1. Rana nigromaculata and Natrix tigrina lateralis were found to be infected naturally by the metacercariae(diplostomula) of F. seoulensis. The metacercarial capsule was round to elliptical in tissue of the intermediate hosts with a long diameter 0.232~0.385 mm. Liberated metacercariae were ovoid with small conical posterior body. Body length measured 0.199~0.312 mm and width 0.153~0.252 mm. 2. The infection rate of R. nigromaculata by the metacercariae ranged from 87% to 100% by area, and the number of the larvae ranged from 3 to 390 by frog. The metacercariae were found in skeletal muscle of frogs, from head to hindlegs. 3. All examined N. t. lateralis were found to be infected by the metacercariae with the range of numbers frome 3 to 35,918. The larvae were collected from all viscera and body segments except for the head of the snakes. However, a great majority of the metacercariae were collected from the stomach. 4. Hematoxylin-eosin stained preparations of frog skeletal muscle and snakes stomach revealed that the metacercariae had no cyst wall of worm origin, but encapsulated by the host tissue. Some of them were found in dilated lymphatic vessels. The larval infection was associated with slight or severe inflammatory reaction even with granuloma formation. By above results, it was concluded that the frog, Rana nigromaculata, was the second intermediate host, and the snake Natrix tigrina lateralis was a paratenic host of F. seoulensis in nature in Korea. PMID- 12902683 TI - Studies On Intestinal Trematodes In Korea: VII. Growth, Development And Recovery Of Fibricola Seoulensis From Experimentally Infected Rats And Mice. AB - This study was carried out to observe the growth and development of Fibricola seoulensis in experimentally infected rats and mice. The metacercariae were obtained from naturally infected Natrix tigrina lateralis which were collected in Hoengseong Gun, Gangweon Do, in August 1982. The results were summarized as follows. 1. Growth and development of worm were not different between the rat and mouse. Body growth was attained within a week in both hosts. The testes differentiated first on the 2nd day after infection, and then ovary on the 3rd day, vitelline follicles on the 4th day, vitelline reservoir on the 5th day. A few eggs appeared in uterus on the 5th day and in feces on the 6th day. The body length grew almost linearly for the first 7 days after infection. The growth of posterior body was more remarkable, and the ratio of posterior body length to that of anterior body increased to 1.099 until 4 weeks after infection. 2. The worms of 4weeks age in rats measured 1.10~1.92 mm long and 0.511~0.696 mm wide. The morphological characteristics of adult worms were identical to the original description of F. seoulensis, though dimensions varied in wider range. 3. The worms were recovered from the rats 12.5% on the day after infecion, and the rate ranged 21.0~59.0% up to 6th day. One week after infection, the rates were 42.6%, 60.9% and 43.9% respectively by the metacercarial group of 200, 1,000 and 2,000 and the rates decreased to 16.2%, 29.1% and 3.1% respectively by the group on the 4th week after infection. Most of the worms were recovered from duodenum, and a part of worms were found in jejunum or ileum in heavily infected rats. 4. The worms of 23.5~63.5% were recovered from mice from 3 to 10 days after infection. A great majority of them was found in duodenum, too. PMID- 12902684 TI - [A Study On The Fine Tegumental Structures Of The Metacercaria And Juvenile Stages Of Clonorchis Sinensis] AB - This study was carried out to observe the chronological tegumental changes of juvenile C. sinensis using scanning and transmission electron microscopes (SEM and TEM). The subjected worms were the excysted metacercaria, and l day, 1 week, 2 week and 4 week old worms. By observation with SEM, the tegument of excysted metacercaria showed many transverse wrinkling or shallow rugae, especially remarkable in anterior half of body, i.e., anterior to ventral sucker. Many spines were arrayed on the whole body surface, and double or triple pointed on anterior half and single pointed on posterior half. The observed sensory papillae were composed of 3 types. The ciliated knob-like papillae (type A in Fujino's classification) were abundant on anterior half, especially around oral sucker. A few plate like elevated papillae (type B) were found on middle part and non ciliated round swellings (type C papillae) were observed around ventral sucker. The tegumental surface of 1 day old worms showed deeper rugae, and the posterior body end was covered with cobble stone-like processes of distal cytoplasm. The spines protruded further and the spines on posterior half of body changed to scale-like ones. The tegument of 1 week old worms became velvety and the spines grew further in length, but the density decreased. The tegumental processes differentiated much finely in the 2 week and 4 week old specimens and the spines or sensory papillae decreased of their relative density. The basic tegumental structures, such as distal cytoplasm containing various granules, vesicles and spines, basement membrane complex, muscular layers and tegumental cells were also monitored in 1 week old worms by TEM. PMID- 12902685 TI - Epidemiological studies of Clonorchis sinensis in lower area of Nag Dong river nearby Busan City in Korea. AB - The infection rates with cercaria and metacercaria of the Clonorchis sinensis in snail and fish hosts, and the prevalence among the inhabitants in the lower Nag Dong river in Busan were investigated in the period from June 1981 to March 1982. The stool examination and E.P.G. count for prevalence and intensity of clonorchiasis were done by cellophane thick smear and Stoll's egg count methods. The prevalence rate of clonorchiasis among inhabitants of 3 towns in Bug Gu, was 38.4% among a total of 1,384 persons examined. By town, the prevalence in Gu Po was 25.4% out of 330 persons examined, 39.3% in 438 examined in Gang Dong and 44.8% out of 616 in examined in Myeng Ji respectively. According to sex, the male(41.9%) showed relatively higher rate than female(35.3%). By age, increase of rate was observed as increase in age. The highest rate was 80.7% in 50~59 years group. The intensity of infection was light: 89.1% of examined cases were below 10,000 E.P.G.; by area 80.9%(Gu Po), 80.2%(Gang Dong), 97.3%(Myeng JI). Out of fresh water fish species collected, 2 species were not infected with metacercaria. The metacercarial detection rate from fish in low Nag Dong river showed 100% in Pseudorasbora parva, Pseudogobio esocinus(41.4%), Hemibarbus labeo(37.1%), Acheilognathus limbata(33.3%) and Erythroculter erthropterus(4.5%) respectively. The mean number of metacercaria per gram of muscle was 21.2 in P. parva, P. esocinus(15.0), H. labeo(11.9), A. limbata(6.3), and E. erythropterus(1.6) respectively. Though 7 area along Nag Dong river were surveyed for Parafossarulus manchouricus, the snails were not found at all areas except for one area, Seo Nam stream; and the cercaria was infected 0.9% out of 708 examined. PMID- 12902686 TI - [Isolation Of Anthelmintic Substance On Clonorchis Sinensis From Tuber Of Bletilla Striata] AB - The present work was done to isolate the anthelmintic substance from the tuber of Bletilla striata which grows Gangweon-do of the Korea peninsula. The dried B. striata tuber was extracted with ethyl ether by a Soxhlet extractor at 40 degrees C for 24 hours, and then the extract was separated into insoluble material (negative wormicidal) and soluble material (positive wormicidal) by petroleum ether solvent. The wormicidal substance was separated in the silica gel column chromatography from the petroleum ether extract, using benzene (90 %) and acetone (10 %) as a solvent. The anthelmintic activity was observed in vitro on the cercaria, the excysted metacercaria and the adult of Clonorchis sinensis. On the silica gel thin layer chromatography with the wormicidal substance in benzene : acetone/90 : 10, five spots were observed in iodine chamber and the spot of Rf. 0.18 value of them appeared the most strong wormicidal effect on the cercaria of C. sinensis. Afterward, a substance which was equivalent to the Rf. 0.18 value spot was isolated from the wormicidal substance by means of silica gel column chromatography using 1 part of ethyl acetate and 6 parts of chloroform carrier, and the excysted matacercaria and the adult of C. sinensis were killed within 14 minutes and 128 minutes in the substance. PMID- 12902687 TI - Larval Trematodes From Semisulcospira Snails In Kyungpook Province, Korea. AB - In an attempt to determine the incidence of the cercarial larvae of trematodes, Semisulcospira snails collected in Judong stream, an endemic area of Paragonimus westermani, in Kyungpook Province, Korea, were examined from 1965 to 1980. The infestation rate for cercariae in the snails was 5.4 per cent, but in 1970 dropped to 4.0 per cent and remained at the same level, 4.2 per cent, in 1975. It then decreased to 3.5 per cent in 1980. Seven species of cercariae, Cercaria incerta(Kobayashi, 1992) Faust, 1924, Cercaria nipponensis(Ando, 1915) Faust, 1924, Cercaria yoshidae(Osafune, 1898) Cort et Nichols, 1920, Centrocestus armatus(Tanabe, 1922) Yamaguti, 1934, Metagonimus species, Notocotylus magniovatus Yamaguti, 1934, and Paragonimus westermani(Kerbert, 1878), and five kinds of undeterminced cercariae were found. Of these, the most freqently emerged cercaria was Metagonimus species, folllowed by Notocotylus magniovatus. The least freqently emerged was Paragonimus westermani, with the rate of 0.04 per cent. The liberation rates in the five kinds of undetermined species varied from 0.04 to 0.15 per cent. In the monthly liberaton rates for cercariae, the highest rate was encountered in May, and decreased in the period from June to November. It then somewhat increased again in December. There was no correlation in the liberation rate for cercariae and length of snails. The results of this study indicate that the infestation of Semisulcospira snails with the cercariae of trematodes has decreased over the fifteen year period from 1965 to 1980. PMID- 12902688 TI - [Changes Of Liver And Diaphragm In Experimental Paragonimiasis] AB - Changes of abdominal organs in early paragonimiasis are described in dogs and cats. Experimental infection was induced by feeding 40~50 and 20~30 metacercariae of Paragonimus westermani to 15 dogs and 15 cats respectively. These animals were killed sequentially starting from 8 days to 120 days after infection. Complete autopsies were carried out in all animals and the tissues were examined microscopically with the main emphasis on changes of the liver and the diaphragm. A definite bout of acute diffuse fibrinoexudative peritonitis with ascites ranging from 5 to 20 ml was present in both dogs and cats at the time of 20 days after infection. This was also the period of the maximum liver damage and diaphragmatic injuries. The peritonitic changes became minimized after 45 days of infection. The liver changes were of two folds. The first one was characterized by numerous pin-point or linear tissue defects on the surface of the lobes, which were often covered with fibrin clots. Some of these defects were impacted by the larvae. These "scratch" marks appeared to be of mechanical effect, and the margins were often banal without a significant inflammatory reaction. The scratch marks were no longer observed in animals examined after 45 days. The second was noted in the Kupffer cells of the liver. A large amount of hemosiderin pigment was noted in the Kupffer cells and became prominent by 20 days through 45 days. Hemosiderin-laden macrophages were found also in the spleens of these animals. Some animals in 30 days group showed the most Kupffer cells heavily loaded with hemosiderin. However, no associated degenerative changes were noted in these cells. The diaphragm was quite unique in early phase of infection in all animals examined. Numerous pinpoint perforative lesions could definitely be seen grossly, and some of these lesions included migrating larval worms inside the tunnels. These lesions were microscopically characterized by tract or tunnel formation with accompanying tissue reaction. The diaphragmatic changes were manifest by 20 days after infection. The tracts of tunnels appeared to have been made by pressure necrosis and surrounding edema, and subsequently were associated with a massive eosinophilic influx and myocytolysis. The eosinophilic abscess and multinucleated giant cells were often observed in the animals of the 30 days group. The diaphragmatic changes seemed to be repaired with or without fibrous scar formation which was occasionally seen in 45 days and later. PMID- 12902689 TI - Therapeutic Trial Of Praziquantal (Embay 8440; Biltricide(R)) On The Dermal And Cerebral Human Cysticercosis. AB - A total of 28 adult cases who were confirmed cysticercosis with or without cerebral involvements were treated with praziquantel at the daily dose of 3 x 25 mg/kg for 3 to 7 consecutive days and was evaluated for tolerance and therapeutic effects in the trials clinically performed.The assessment of drug efficacy of praziquantel in the dermal cysticercosis was made by comparing the numbers of cysticercus nodules and histopathological findings of the biopsied parasites by means of light, scanning and transmission electron microscope. In the cerebral cysticercosis, the assessment was considered by the frequency of the episodes of convulsive seizure before and after treatment with praziquantel and by the findings of the disapearance or decreased densities of the lesions in C.T. scan in comparison with those of before and after treatment. The results were as follows: 1. The cysticerci in the subcutaneous tissues began to disappear within one month of drug administration of 3 x 25 mg/kg praziquantel over 3 to 7 days. Within 3 to 6 months most of the cysticerci had disappeared, although in some case a small number of cysticercus nodules remained even one year after treatment. 2. Histological observation of the cysticerci biopsied at different times during the course of treatment revealed that morphological changes were already taking place within two weeks after the treatment. At the early stage of the treatment, small vacuoles were scattered along the basement layer in the tegumental syncytium of the scolex and neck regions. In the scanning electron microscopic observation, marked surface changes were present in the neck region with many bleb-like structures formed by the bursting of the large vacuoles in the tegumental syncytium. In the specimens biopsied at 2 or 5 weeks after treatment, the degenerations and necrosis of the tegumental syncytium were seen in all parts of cysticercus. 3. In 12 cases of cerebral cysticercosis treated with praziquantel at the daily dose of 3 x 35 mg/kg for 3 or 4 consecutive days, there were no ceasing of the convulsive seizures during the 6 months follow-up. Among them 9 cases were given again the same doses of the drug for 4 or 7 days. In 7 of 9 cases, no more convulsive seizure was experienced over one or two years after the second time. At the same treatment the lesions of the brain C.T. scan disappeared, decreasd in size or calcified after treatment. In other 3 cerebral cysticercosis cases, complete cure was also obtained after the oral medication of praziquantel at the daily dose of 3 x 25 mg/kg for 7 consecutive days. 4. In the treatment of cerebral cysticercosis with praziquantel, it was found that the concomitant oral medication of dexamethasone during the course of treatment was effective for preventing and minimizing the side-effects. PMID- 12902690 TI - A Human Case Of Ectopic Fascioliasis In Korea. AB - A case of ectopic fascioliasis involving cecum and ascending colon was reported. This case was a 19-year old Korean female who had a history of postprandial aggravation of intermittent abdominal pain of 3 weeks duration. She was presented with an abdominal mass in the right upper quadrant, for which an operation(right hemicolectomy) was performed. Whole cecum and ascending colon showed multifocal tunnel-like necrotizing granulmas with central cavitations. Numerous Charcot Leyden crystals were seen along tracts of the worm. The lesion was distributed throughout the entire thickness of the wall. Parasitological examination confirmed the worm found in the intestinal wall to be juvenile form of Fasciola species. Possibilities were discussed on the localization of the fluke and the mode of infection of present case. PMID- 12902691 TI - Clavate Cells Of Epidermis In Cyprinus Carpio Nudus With Reference To Its Defence Activity To Clonorchis Sinensis. PMID- 12902692 TI - Therapeutic field trial with praziquantel (Biltricide(R)) in a rural population infected with Clonorchis sinensis. AB - The present study was undertaken to determine the acceptablility, tolerance and effectiveness of praziquantel in a rural population infected with Clonorchis sinensis and to suggest the suitable dosages of praziquantel for the field use on a large scale. A total of 121 patients with proven C. sinensis infection were treated by two dosages with praziquantel at a single dose of 40 mg/kg bwt and 2 doses of 30 mg/kg bwt in a single day. A single dose of 40 mg/kg bwt were given to 60 patients and 2 x 30mg/kg bwt were given to 61 patients. Follow-up examinations were carried out at about 30 and 60 days after treatment. Two months after therapy, 13(21.7%) of 60 patients who received a single dose of 40 mg/kg bwt were cured completely. Among these cured patients, 9(75.0%) out of 12 cases of light and only 4 out of 48 cases of moderate or heavy infection groups were cured. But in the non-cured cases the overall egg reduction rate was 89.1%. On the other hand, 36(59.0%) out of 61 patients treated with 2 x 30 mg/kg bwt were cured at 60 days after treatment. Among these cured patients, all of the 13 cases of light infection and 18(69.2%) out of 26 cases of moderate and 5(23.8%) out of 21 cases of heavy infection groups were cured completely at 60 days after therapy. However the overall egg reduction rate was 95.2% in the non-cured cases. Praziquantel is well tolerated and side effects consist particularly of mild and transient headache, dizziness and abdominal discomfort, etc. However there was no difference in regard to frequency and intensity of untoward side effects between the two dosage groups. The results obtained in this study suggest that a single dose of 40 mg/kg bwt for light infection, 2 x 30 mg/kg bwt for moderate infection and 3 x 25 mg/kg bwt for heavy infection groups will be recommended for the field use on a large scale. PMID- 12902693 TI - [Localization and isolation of anthelmintic activity on Clonorchis sinensis from stem cortex of Machilis thunbergii] AB - The present work was to localize and isolate the anthelmintic substance from stem cortex of Machilis thunbergii which grows in all parts of the Korea peninsula. The dried M. thunbergii bark was extracted with a solvent mixture (Methanol: Ethyl acetate/1:1) by a Soxhlet extractor at 70 degrees C for 24 hours, and then the extract was separated into solid matter (negative wormicidal) and benzene extract (positive wormicidal) by benzene solvent. The anthelmintic action was observed on the cercaria, the liberated metacercaria and the adult of Clonorchis sinensis. On the silica gel thin layer chromatography with a spot of benzene extract in benzene solvent, nine spots were observed in iodine chamber and the spot of Rf. 0.16 value of them appeared the most strong wormicidal effect on the cercaria and the liberated metacercaria of C. sinensis. Afterward, a 0.3 g colourless solid substance which was equivalent to the Rf. 0.16 value spot was isolated from the benzene extract of 3 kg M. thunbergii stem cortex by means of silica gel column chromatography using benzene carrier, and the adult of C. sinensis was killed within 6 minutes by 0.05 percent solution of the substance. PMID- 12902694 TI - The Detectability Of Helminth Eggs From Feces By Cellophane Thick Smear Technique. AB - The efficacy of the cellophane thick smear(CTS) technique was reassessed for detecting helminths eggs such as Ascaris lumbricoides, Trichuris trichiura and Clonorchis sinensis, in comparsion with the formalin-ether(F-E) one, with repeated smears per case. From the fecal samples of 411 ROK Army soldiers, a total of 3,288 smears were made and examined; 4 by CTS and 4 by F-E techniques. The expected egg detectability with one, two, three or four smears by each technique was mathemativally calculated. The results were as follows: 1. When the results of all 8 smears per case were integrated, the number of cases with Ascaris egg were 54(13.1%), Trichuris 125(30.4%) and Clonorchis 33(8.0%). Some other kinds of helminths eggs and protozoan cysts were found from a few cases each. 2. The calculated detectability of Ascaris eggs by CTS technique was fairly high(95.8~96.3%) with 1-4 smears, while that by F-E technique was not so high(73.1~88.9%). 3. In case of Trchuris eggs, the egg detectability by CTS technique with 1-4 smears was a little lower(62.2~86.4%) than that by F-E technique(73.4 ~ 91.2%). 4. The detectability of Clonorchis eggs with 1-4 smears by CTS technique was the lowest(44.7~60.6%) among the three helminths as well as much lower than that by F-E technique(67.4~87.9%). PMID- 12902695 TI - The Effects Of Common Salt On Aedes Togoi And Filarial Infection. AB - The effects of salt on the development of Aedes togoi and the infecton of Brugia pahangi were studied. Salt concentrations over 7.2% completely got rid of pupae and adults. The salt concentrations 0% to 3.6% reduced pupal development from 86.6% to 72.8% and adult from 70.3% to 53.3%. B. pahangi infestations and infection rates were also reduced in the mosquitoes reared at salt concentrations of 1.8% and 3.6%. Trends of retardation in the filarial development were observed when the salt concentrations were increased. PMID- 12902696 TI - [Study On Metagonimus Yokogawai (Katsurada, 1912) In Korea: VI.The Geographical Distribution Of Metacercarial Infection In Sweetfish Along The East And South Coast] AB - This study was carried out to outline the geographical distribution of the sweetfish which were infected by the metacercariae of Metagonimus yokogawai from August to October in 1979. The infection status was examined both by the muscle compression method and by digestion technique. The result was summarized as follows: 1. A total of 197 sweetfish, Plecoglossus altivelis was collected, 125 fish from 5 streams along the East Coast and 72 fish from 4 streams along the South Coast. 2. Total 125(63.5 %) fish were infected by the metacercariae of M. yokogawai. The rate and intensity of infection in the south coastal areas were much higher than those in the east coastal areas; the rate was 42.4 % in the East and 100 % in the South, mean number of metacercariae per fish was 721 in the East and 14,308 in the South, and mean number of metacercariae per gram of flesh was 43 in the East compare with 904 in the South. 3. The sweetfish infected by the metacercaria of M. yokogawai were newly observed in 5 foci, i.e., Yeongogcheon in Myeongju Gun and Namdaecheon in Gangneung Si, Gangweon Do and Wangpicheon in Uljin Gun, Gyeongsangbug Do in the East, and a branch of Namgang in Euiryeong Gun and Sinhyeoncheon in Geoje Gun, Gyeongsangnam Do in the South. The rate of metacercarial infection was 4.1 % in Yeongogcheon and 100 percent in others. The number of metacercariae in a fish ranged from 9 to 45,018 and the number of metacercariae per gram of flesh was the lowest in Gangneung Namdaecheon as 19 and the highest in Geoje Sinhyeoncheon as 1,237. Summarizing these findings with previous records, M. yokogawai might be prevalent in the streams along the East Coast southern from Myeongju Gun, Gangweon Do and along the South Coast in Korea. PMID- 12902697 TI - [Toxoplasma antibodies by indirect latex agglutination tests in St. Mary's Hospital patients] AB - A total of 421 patients hospitalized in St. Mary's Hospital were examined by indirect latex agglutination test in order to evaluate the Toxoplasma antibody in Korean from June to August 1981. The test sera of the patients were obtained from each age group by random sampling. 1. The 421 samples of test sera showed negative in 153, 1:2 in 157, 1:4 in 59, 1:8 in 27, 1:16 in 7, 1:32 in 9, 1:64 in 2, 1:128 in 4 and 1:256 in 3 cases, respectively. 2. The positive rate of Toxoplasma antibody was 4.3 percent in this sample when indirect latex antibodies of 1:32 or higher were regarded as positive. 3. The titers of positive Toxoplasma antibodies were increased by age. PMID- 12902698 TI - [The Status Of Intestinal Protozoan Infections In Inhabitants Of Gangweon-Do, Korea] AB - To evaluate the status of intestinal protozoan infections in inhabitants of Gangweon-Do, Korea, a total of 1,310 stool specimens (male 669, female 641) was collected from 2 cities and 3 counties. They were examined routinely 1 time by the method of formalin-ether sedimentation technique. The results were as follows: 1. The positive rate for any kind of the intestinal protozoan cysts was 8.9 %. 2. A total of 6 kinds of the intestinal protozoan cysts were detected. The prevalence rate of each protozoa were; E. histolytica 0.8 %, E. coli 7.6 %, E. nana 1.4 %, I. butschlii 0.2 %, G. lamblia 0.5 % and C. mesnili 0.5 %. 3. Sogcho city showed the highest positive rate as 15.2 %, Myeongju county was the next as 11.3 % and Weonju-city showed the lowest positive rate as low as 3.9 %. 4. By age, the highest positive rate was found in 20-29 age group(12.4 %). Female (9.5 %) showed a slight higher positive rate than male (8.4 %). PMID- 12902699 TI - [A Survey On The Prevalence Of Intestinal Protozoan Cysts In Jeonlanam Do, Korea] AB - A survey on the prevalence of intestinal protozoan cysts was conductcd in Jeonlanam Do (Province), Korea in 1981. A total of 4,116 specimens was collected from 2,035 male and 2,086 female inhabitants in 4 urban and 7 rural areas. Each case was selected randomly to represent 1,000 inhabitants referred to the census in 1980. Formalin-Ether sedimentation technique was employed to concentrate the cysts and one Lugol's iodine stained smear was examined for each case. The results were summarized as follows: 1. The cases who passed out protozoan cysts of any kind were 9.1 % out of total 4,116 examined, and cumulative positive rate was 11.0 %. Among the positive cases, 16.5 % were infected by two or more species of protozoa. 2. Total 5 species of cysts were detected and the positive rates were as follows; Entamoeba histolytica 1.4 %, E. Coli, 4.9 %, Endolimax nana 2.5 %, Iodamoeba butschlii 0.4 % and Giardia lamblia 1.9 %. 3. The cyst positive rates varied greatly by the surveyed area. The positive rate in rural area was 11.6 % compared with 5.7 % in urban area. The highest rate was found 14.5 % in Gurye Gun (County) and the lowest 3.6 % in Mogpo Si (City). 4. Female showed much higher positive rate than male as 11.8 % compared with 6.4 % and the cumulative rate 14.2 % and 7.6 % respectively. By age group, the rate was highest as 13.9 % in 40~49 years and next 12.5 % in over 60 years group. The cyst prevalence rate diminished remarkably in Jeonlanam Do compared to previous records. However, the finding that 20.2 % of females of 40 years were the carrier of protozoan cysts was an important information in public health point of view. PMID- 12902700 TI - A Case Of Niclofolan (Bilevon(R)) Intoxication. AB - The authors reported a case of niclofolan intoxication occurred during the trial of clonorchiasis treatment. The case, a 15 years old Korean schoolboy, took niclofolan(Bilevon(R)) of total 473 mg(11 mg/kg) in 11 divided doses during 20 days. And the case suffered from neurologic symptoms such as severe headache, dizziness, nausea, vomiting, blurred vision, papilledema, retinal hemorrhage, an epsiode of seizure attack and elevated intracranial pressure, and hepatotoxic symptoms such as hepatomegaly, increased serum transaminases, and shoulder pain, excessive sweating and weight loss. Therapy was concentrated to the management of the elevated intracranial pressure. Hepatotoxic manifestations subsided within one month. The clinical signs related to elevated intracranial pressure persisted two months. Body weight regained after 2 months. And the symptoms of headache, dizziness and vomiting were complained intermittently until 4 months after onset. However, no subsequent clinical problems related with this episode has been noted until this record. PMID- 12902701 TI - A Case Of Systemic Paragonimiasis With Ovarian Involvement. AB - The authors reported a case of ectopic paragonimiasis in a 40 year old Korean housewife who came to the hospital because of abdominal mass and pain. She was proved to have multiple parasitic granulomas involving lung, uterus, uterine tube and subcutaneous tissue. The ovary was also the site of parasitism. The past history revealed an episode of ingestion of fresh water crabs soaked in soybean sauce which were obtained in a market of Seoul City 2 years prior to the present illness. The patient was treated for tuberculosis for a year without confirmation of etiologic agent. After surgical removal of all abdominal and most of pelvic granulomas, the patient was treated with praziquantel 25 mg/kg t.i.d. for 3 days, and was judged to be cured after 4 months. PMID- 12902702 TI - [A Case Of Human Thelaziasis In Korea] AB - This paper deals with the 11th case of human thelaziasis in Korea. The patient was a 25-year-old woman with complaints of foreign body and itching sensation of rihgt eye, who had lived in Seoul. The slender milky-white nematodes extracted from lower conjunctival sac, one male and one female in respect, were confirmed as Thelazia callipaeda Railliet and Henry, 1910. PMID- 12902703 TI - Parasitic amebae in Korea. AB - This review dealt with biology, host-parasite relationship, symptomatology, epidemiology, diagnosis, treatment of Entamoeba histolytica infection and free livng amoeba. PMID- 12902704 TI - [Anthelmintic Effects Of Single Doses Of Fenbendazole And Oxantel-Pyrantel Pamoate To The Intestinal Nematodes] AB - The present study was undertaken to evaluate the anthelmintic effects of fenbendazole against intestinal nematode; Ascaris lumbricoides, hookworm and Trichuris trichiura, and to compare the efficacy in fenbendazole, oxantel pyrantel pamoate and placebo by means of double blind method. Out of 114 subjects harbouring Ascaris lumbricoides, hookworm and Trichuris trichiura, 36 cases were treated with single dose of fenbendazole, 38 cases with oxantel-pyrantel pamoate, and the remaining 40 cases had received the placebo. The results were as follows: 1. In the group treated with fenbendazole (30~50 mg/kg), the cure rates were 83.9 per cent in 31 subjects with Ascaris lumbricoides and 83.3 per cent in 18 subjects with hookworm, and only 28.6 per cent in 28 subjects with T. trichiura respectively. 2. In the group treated with a single dose of oxantel-pyrantel pamoate (10 mg/mg), the cure rates were 96.7 per cent in 30 subjects with A. lumbricoides, 95.2 per cent in 21 subjects with hookworm, and 54.6 per cent in 33 subjects with T. trichiura. Egg reduction rate was 85.7 per cent in T. trichiura cases. 3. On the other hand, the egg negative conversion rates in placebo group were 9.7, 8.3 and 33.3 per cent in Ascaris, Trichuris and hookworm infections respectively. 4. The above results showed that fenbendazole was highly effective against Ascaris and hookworm. However, incomparisom with oxantel-pyrantel pamoate, fenbendazole was less effective in regards of A. lumbricoides, hookworm and T. trichiura infections. PMID- 12902705 TI - [Agar-Gel Precipitin Reactions In Experimental Paragonimiasis] AB - In an attempt to investigate the sensitivity of immunodiagnosis in cats experimentally infected with Paragonimus westermani, agar-gel precipitin reaction were studied. Metacercariae of P. westermani were administered to cats in various doses(2~100 metacercariae per cat) and antisera were obtained at an interval of a week. 1. Precipitin bands appeared in homologous antigen-antibody in experimental paragonimiasis between 3 and 5 weeks after infection in all the cats. 2. Almost all the cases in which a large number of worms were detected, showed strong reactions as revealed by deeply stained bands. 3. Precipitin reactions did not necessarily parallel with the number of worms detected. This may be attributable to the individual difference of a cat's conditions. 4. Very weak precipitin reactions were noticed between Clonorchis antigen and Paragonimus antisera of cats, but no reactions were noticed between Paragonimus antigen and Clonorchis antisera of cats or rabbits. PMID- 12902706 TI - Study On The Pathology Of Metagonimiasis In Experimentally Infected Cat Intestine. AB - To study the basic pathological changes of small intestine in metagonimiasis, light- and electron microscopic studies were made, using a total of 21 cats which were experimentally infected with metacercariae of Metagonimus yokogawai. The metacercariae were obtained from naturally infected sweetfish (Plecoglossus altivelis) by digestion technique. The cats were divided in control, light infection(10,000 metacercariae infected) and heavy-infection(50,000 metacercariae infected) groups. Cats were killed at the 5th, 10th, 15th day, and 4th, 8th and 10th week after the infection. And the small intestine was prepared for the study. Pathological studies comprised gross examination, worm distribution pattern, light microscopic examination and both transmission and scanning electron microscopic examinations. The results obtained were summarized as follows. 1. Gross morphologic changes were the most marked during the first 2 weeks after infection. The gross abnormalities were severer in the heavily infected animals. The changes were dryness and listlessness of serosal surface due to dehydration, mushy and/or watery intestinal content, effacement of transverse nodes and enlargement of mesenteric lymph folds and Peyer's patches. After 4 weeks of infection, these changes became less marked showing a tendency to return to normal. 2. The sectioned flukes were distributed from duodenum to proximal ileum. However, individual variation was marked in distribution. In the heavy-infection group, the locality of parasitism tended to extend more distally. 3. The locality of M. yokogawai in the intervillous space was mostly in the lower most portion of intervillous space, where they compressed and eroded epithelial cells probably due to mechanical damage to the structure. Very rarely the worms were found in lumen of Lierberkuehn's crypt, and reaching, in two occasions, into proprial lymphoid tissue. 4. Light-microscopically the lesion was restricted in mucosa: Early mucosal changes were shortening, blunting, fusion, and thickening of the villi, crypt hypertrophy with consequent decrease of villus/crypt ratio, as well as stromal changes of edema, capilliary ectasia and marked inflammatory cell infiltration of lymphocytes and plasma cells. Goblet cells were markedly reduced in number as with depletion of its cytoplasmic content. In the later stages of infection, mucosa restored its normal configuration in spite of persistent parasitism of the worms. 5. At the infection stage of 5-15 days, there was significant shortening of the microvillous height with varible destruction of glycocalyx in electron microscopic examination. With lapse of infection time, microvilli became to restore the normal pattern. With these morphological changes, it appears that diarrhea in experimental metagonimiasis would be related to the decrease of absorptive surface of the small intestine particularly in the early phase of infection. The significant changes seen in villi and microvilli might be due to massive intrusion or invasion of Metagonimus worms into the crypts, causing direct mechanical and possible host-immune response to the small bowel mucosa. PMID- 12902707 TI - Studies On Parasitic Helminths Of Korea 5.Survey On Intestinal Trematodes Of House Rats. AB - A study was carried out to evaluate the house rats in southern Korea as reservoir host of intestinal trematodes, in 13 different areas: 7 in inland and 6 in brackish-water zones, during the period from August 1980 to August 1981. A total of 170 house rats was captured; 101 rats from inland and 69 from brackish-water zones. They consisted of 129 Rattus norvegicus, 22 R. rattus rufescens, and 19 unidentified. The infection status of the rats were as follows: 1. Total 29 rats (17.1%) were infected by one or more kinds of intestinal trematodes; 27 from inland and 2 from brackish-water zones by areas. Rats in inland were more heavily infected. 2. The intestinal trematodes collected from rats were identified as Echinostoma hortense, E. cinetorchis, Plagiorchis muris, Fibricola seoulensis, Clonorchis sinensis and Metagonimus yokogawai. 3. As a reservoir host, the rats were very important in Echinostoma hortense and E. cinetorchis infection. Rats infected with these trematodes were found widely in southern Korea, and its infection rates were the highest among the discriminated flukes. 4. In this study, no rats were found to be infected with Heterophyes and Pygidiopsis even in endemic areas. The negligible importance of rats as reservoir host of these heterophyid should be reevaluated in the future. PMID- 12902709 TI - Application of micro-ELISA in serodiagnosis of Human paragonimiasis. AB - To observed whether the specific IgG antibody test using ELISA was useful in diagnosis of presently ill patients of paragonimiasis, a total of 95 sera were tested. The sera were collected from 21 egg positive cases, 8 from positive reactors of intradermal test, 7 from Clonorchis infected, 9 from other parasitic diseases and 50 from apparently non-infected cases. By the result, the sensitivity of the test was 86% and the specificity was 100%. There were no cross reactions between Paragonimus antigen and other parastic infections. Specific IgG antibody test by micro-ELISA was concluded to be useful for mass screening of the presently ill paragonimiasis in the field. PMID- 12902708 TI - An Epidemiologic Study On Clonorchiasis And Metagonimiasis In Riverside Areas In Korea. AB - A study was carried out to figure the cases and to observe the endemic status of clonorchiasis and metagonimiasis in endemic areas in Korea. Total 13,373 inhabitants living in 40 villages along 7 main rivers and 9 small streams were subjected to stool examination. They were selected randomly among the riverside population. Their specimens were examined both by cellphane thick smear method and Stoll's egg countung technique. This study was performed during the period from May 1979 to April 1980. The results obtained are as follows: 1. The egg positive rate of any kind of helminths was 58.7% out of 13,373 examned cases, and the egg positive rates by each helminth were; Clonorchis sinensis 21.5%, Metagonimus yokogaqai 4.8%, large type Metagonimus eggs 0.4%, Ascaris lumbricoides 22.9%, Trichuris trichiura 35.2%, hookworm 0.2%, Taenia sp. 0.5%, Hymenolepis nana 0.07%, Paragonimus westermani 2 cases, Fasciola sp. 4 cases, Echinostoma sp. 1 case and Hymenolepis diminuta 1 case respectively. 2. Many endemic foci of clonorchiasis were revealed along the 7 major rivers. The egg positive rates of each river basin differed from each other significantly; Nagdonggang 40.2%, Yeongsangang 30.8%, Seomjingang 17.3%, Hangang 15.7%, Tamjingang 15.9%, Geumgang 12.0% and Mangyeonggang 8.0%. The cases of clonorchiasis were estimated in range 830,000 to 890,000 in riverside areas of the 7 rivers. By grading the infection intensity, 64.7% was in Grade I(EPG 0 900), 28.6% in Grade II (EPG 1000-9,900), 5.5% in Grade III (EPG 10,000-29,900) and 1.3% in Grade IV (EPG over 30,000). The proportion of the cases in Grade III and IV was 6.8% among positive cases. Therefore 60,000 cases at least were regarded to suffer from it clinically. Males of 30-60 years of age showed higher positive rate and heavier burden of infection. This makes clonorchiasis more important socially because the patients lose their social productivity. 3. A few endemic foci of metagonimiasis were detected newly by egg detection; Samcheong 28.5% egg positive rate, Uljin 21.3%, Yeuongdeog 46.3%, Milyang 6.7%, Yeongil 9.2% and Geoje 18.2%. The mean EPG values were in range of 320-7, 120 by the focus. The egg positive rate and proportion of EPG Grade varied greatly by the area, and mean proportion of the positive cases were 69.7% in Grade I, 24.1% in Grade II, 5.0% in Grade III and 1.2% in Grade IV. Males of 30-60 years were infected in higher rate also. The large sized eggs of Metagonimus were found also in upper basin of Hangang and Geumgang mainly. They were regarded as eggs of M. takahashii which is mediated by the cyprinid fishes. Its significance should be studied further. Clonorchiasis and metagonimiasis should be realized as important public health problems in Korea by their wide distribution, high prevalence rate and heavy infection intensity. Comprehensive measures against them are needed urgently. PMID- 12902710 TI - Infestation With Larval Trematodes From Fresh-Water Fish In Natural And Fish Breeding Ponds. AB - The present study was undertaken to determine the pattern of infestation with the encysted larvae of digenetic trematodes in freshwater fish collected in both the natural and fish breeding ponds in Kyungpook Province, Korea. The fish collected in the two natural ponds, Nass-chil, and Sa-il, and three fish breeding ponds, Deok-dong, Mo-gok, and Seongryu-tunnel, were examined from July 1980 to August 1981. The data were compared with the results obtained from freshwater fish caught in the river Kumho by Hwang and Choi(1980). Ten species of fish caught in the ponds were examined for the presence of the larval trematodes. Seven species of the larvae, Clonorchis sinensis, Cyathocotyle orientalis, Echinochasmus species, Exorchis oviformis, Metacercaria hasegawai, Metagonimus yokogawai, and Metorchis orientails and two kinds of undetermined cyst A and B were found. The infestitation rate for Clonorchis sinensis metacercaria in the Korean shiner, Gnathopogon atromaculatus collected in the natural ponds was higher than the rate in the river Kumho by Hwang and Choi(1980), but lower in the fish collected in the breeding ponds. By contrast, the infestation rate in the flat bitterling, Paracheilognathus rhombea in ponds was higher, while there was no change of the rate in the southern top-mouthed minnow, Pseudorasbora parva. The intensity of infestation, expressed as the average number of Clonorchis sinensis metacercaria per gram of flesh, were lower than in the bullhead, Coreobagrus brevicorpus, and Gnathopogon atromaculatus in the ponds in 1981. The intensity was higher in Paracheilognathus rhombea, but no fluctuation in the intensity of infestation in Pseudorasbora parva was observed. The infestation rates and densities for the encysted larvae of digenetic trematodes except for Clonorchis sinensis varied from fish to fish. However, it was found that the rate and intensity of the larval trematodes from all the fish in the ponds was lower than from those in the rivers in Kyungpook Province, Korea. PMID- 12902711 TI - A Case Of Taenia Saginata Infection Involving Gallbladder And Common Bile Duct. AB - A 77-year old man living in Kyunggi-Do, Korea was surgically treated at Seoul National University Hospital on April 1981, because of acute abdomen. At laparotomy, a 1.77m long adult Taenia saginata was found both in gallbladder and in common bile duct to cause acute gangrenous cholecystitis. The relevant literature were reviewed and possible mechanisms of the disease were discussed. PMID- 12902712 TI - Pharyngostomum cordatum (Trematoda: Alariidae) collected from a cat in Korea. AB - Ten adult worms of Pharynogostomum cordatum were recovered from a cat instesine purchased in August 1980 at the Namdaemun Market, Seoul, Korea. It is the first case report of cat pharyngostomiasis in Korea. PMID- 12902713 TI - [A Comparative Analysis Of Various Parts Of Ascaris Suum With Respect To Their Protein Composition] AB - For the purpose of making a comparative study of protein compositions in Ascaris suum by sexes and body parts, extracts were prepared from whole bodies, body walls, genital organs, digestive organs and body fluid, of both sexes. And electrophoretic analysis was conducted using polyacrylamide slab gel in the presence of sodium dodecyl sulfate. The results were as follows: 1. In this study, protein bands of each part were separated in the largest number and most clearly under 8%~12 % (10 %) gradient gel condition. The number of bands by body parts was 43 in body walls, 51 in genital organs, 47 in digestive organs, and 34 in body fluid. 2. When examined in terms of sex, the number of bands of whole body was 38 in females and 35 in males. 3. The electrophoretic patterns of body wall protein were in most cases similar with those seen in digestive organs. And the band with a molecular weight of 72,900 was unique to the body wall, and the 122,000 MW band was unique to the female body wall. 4. In genital organ extracts, large molecular weight proteins (more than 80,000) were more frequently met. The molecular weights showed some differences between the two sexes. Of the bands, those having molecular weights of 119,700, 100,500, 88,500 and 86,100 were unique to the female genital organs. On the other hand, the male genital organs showed unique bands having molecular weights of 87,100, 82,800, and 15,500. An unique band common to the genital organs of both sexes was one having 49,300 MW. 5. In the digestive organs evenly distributed protein bands of molecular weights of l0,000~120,000 were observed. The band having 59,800 MW was unique to the digestive organs. The number of bands obtained from body fluid was comparatively small, and the number of bands having less than 30,000 MW was 7, accounting for 55 % of the total protein amounts. The bands having 47,600 MW and 31,400 MW were unique to body fluid. PMID- 12902714 TI - [Analysis Of Protein Components At Varioue Stages Of Clonorchis Sinensis] AB - In this study the authors examined the protein components at various stages of Clonorchis sinensis, and those of tegument and metabolite of adult Clonorchis by using SDS-polyacrylamide gel electrophoresis and immunodiffusion. The following results were obtained: 1. The protein components of C. sinensis were gradually changed during its development. A considerable change occurred during the initial 7 days after the metacercarial infection. 2. Two bands of protein of about 97,000 molecular weight (MW) and 178,000 MW were unique to excysted metacercaria of C. sinensis. Other 2 bands of protein of 23,000 and 25,000 MW which were absent in metacercariae, might be associated with the development of sex organs in adult. 3. In the metabolite, some components of tegumental proteins were detected. And this tegumental protein components in metabolite seems to be the major antigenic components reacting with infected rabbit antiserum by immunodiffusion. 4. Twenty bands of protein were detected in the isolated adult tegument. Among them 6 bands were in 97,000~65,000 MW, 3 bands in 56,000~53,000 MW and 5 bands in 37,000~30,000 MW. On the other hand, in metabolic products of adult C. sinensis, 17 bands were detected. PMID- 12902715 TI - Morphological Observation Of Enterobius Vermicularis Expelled By Various Anthelmintics. AB - When enterobiasis cases were treated with anthelmintics only for one time, the interval to recurrence was variable by different drugs used. And this phenomenon is supposedly connected with biological or developmental cycle of the worm and the consequent efficacy of the different anthelmintics. This study was undertaken to confirm this fact by studying the expelled worms morphologically to correlate the anthelmintics efficacy and stage of worm development in Enterobius vermicularis. A total of 131 children in 3 orphanages was examined by 4 anal swabs (mean positive rate, 80%). They were randomized into 5 experimental groups. Each group was treated with placebo, mebendazole, pyrantel, pyrvinium, and piperazine (70 mg/kg, single dose) respectively. After treatment, all stool were collected for 3 days to get the expelled Enterobius. A total of 6,165 pinworms was studied under the microscope. The sex was discriminated and the length was individually measured. A number of male pinworms was collected in all groups. Females of 2~11 mm in length were also collected in 5 groups. However, significantly larger number of short females was observed in mebendazole group compared with other groups. Twenty-one days after the first treatment, all children were again treated with mebendazole. Once more stool were examined. A total of 1,853 worms was collected. In mebendazole group, there were no females longer than 8.74 mm in the second treatment. In pyrvinium group, 8.31mm in lenght was the longest for female. However in control, pyrantel and piperazine groups, females of 2~11 mm in length were collected. From above results, one could conclude that the removing ability of mebendazole and pyrvinium was satisfactory for the worms in the early stage of development in Enterobius. Pyrantel and single dose of piperazine showed less effective in worm reduction ability especially on those at the early stages. PMID- 12902716 TI - Clinical Evaluation Of Praziquantel(Embay 8440; Biltricide(R)) In The Treatment Of Paragonimus Westermani. AB - A total of 52 paragonimiasis patients was treated with praziquantel at three dose levels: 21 patients received 3 x 25.0 mg/kg bwt on a single day, 21 patients were treated with 3 x 25.0 mg/kg for 2 consecutive days and 10 patients were treated with the same dose for 3 consecutive days. Follow-up examination were carried out at monthly up to 4 months (120 days) after treatment. Fifteen (71.4%) out of 21 patients who received the drug 3 x 25.0 mg/kg bwt on a single day were parasitologically cured. Eighteen (85.7%) out of 21 patients who received 3 x 25.0 mg/kg bwt for 2 consecutive days were also cured. Six and 3 of uncured cases in each above groups were treated again with doses of 3 x 25.0 mg/kg bwt for 2 or 3 consecutive days. Two (consisting each one in each group ) of nine retreated cases were failed in parasitological cure. Therefore the overall cure rates of 95.2 % (20 out of 21 cases) in each group were finally obtained. On the other hand, in 10 patients who received 3 x 25.0 mg/kg bwt for 3 consecutive days, complete cure was obtained at 4 months follow-up examinations. Praziquantel is well tolerated and side effects consist particularly of mild and transient headache and dizziness. There was no great difference between the three dosage groups. Extended hematological and biochemical tests, and urinalysis, revealed no abnormal findings which could be related to the compound after therapy. The disappearance of precipitating bands of immunoelectrophoresis together with the disappearance of abnormal shawdows in chest X-ray after treatment gave a potent proof on assuring the cure of paragonimiasis. PMID- 12902718 TI - [Electron Microscopical And Histochemical Studies On The Epicuticle Of Echinorhynchus Gadi (Acanthocephala)] AB - For the purpose of observing ultrastructure of the epicuticle of Echinorhynchus gadi, the present electron microscopical studies had been made. Also the histochemical methods of Morwy, Bauer, Smith, Lison, Taft, and those of lead and uranyl acetate had been used in order to see the distribution of glycogen, mucopolysaccharides, lipid and nucleic acid in the cuticle of Echinorhynchus gadi. The results obtained by the above studies were as follows: 1. Glycogen, mucopolysaccharides were found in the outermost, middle and inner layers, especially abundant in the middle layer of the cuticle. 2. Lipid was found in the middle and inner layer, and it was found abundantly around the lacunal canal in the cuticle. 3. Nucleic acid was found around the lacunal canal in the middle layer, and also distributed in the cell nucleus of inner layer in the cuticle. 4. Electron microscopically, the cuticle of Echinorhynchus gadi had three outer layers, being outermost, middle and inner ones. The outermost layer was medium electron dense, composed with plasmalemma and filaments. The middle layer was homogeneous one which was electron pale. The inner layer, which was electron dense, consisted of felt layer and radial layer. The electron dense glycogen, lipid granules were distributed in radial layer. PMID- 12902717 TI - [Fluctuation Of The Egg Production Amounts According To Worm Burden And Length Of Ascaris Lumbricoides] AB - This study was undertaken to observe the fluctuation pattern of E.P.G. (eggs per gram of feces) according to the number of infected worms and developmental status of Ascaris lumbricoides. The cases were selected from 2,109 inhabitants of Hwasung Gun, Kyunggi Do, Korea, who had received pyrantel treatment in dose of 10 mg/kg during April 1977-May 1980 and who were cooperative enough to collect whole day stool specimens for 2 days. Among them, 602 cases who had been infected with 1-5 worms were subjected to analysis. Egg examination was undertaken by means of cellophane thick smear technique and the amount of produced eggs was checked by Stoll's egg counting technique. The Ascaris worms were collected from stool specimens, counted in number in each case, and each worm was measured by length. The results could be summarized as follows: 1. According to the increase of worm burden per case from l to 5, the egg detection rate increased from 33.4 % to 95.7 %. And the same pattern was observed when the only burden was considered. 2. There were ample variations in the individual E.P.G. counts even among the cases with same burden, although the mean E.P.G. counts among the cases were quite proportionately increased from 842 to 3,467 according to the number of female worms, 1-4. 3. However, from the distribution pattern of the number of each grade of E.P.G. count, it was revealed that the lower the number of females the more cases were with low grade of E.P.G. 4. It was observed that the eggs could be detected after the female worm grew longer than 12.6 cm and the amount of eggs increased until the worm became about 25.0 cm, where a plateau was made in E.P.G. 1,300~1,400, and decreased significantly after 27.5 cm. And yet, wide variations in E.P.G. counts were noted even among the worms of similar growth. Accordingly, the E.P.G. value had better be used as a useful index to estimate the intensity of Ascaris infection and to compare the intensity between two communities, rather than be used to estimate the individual worm burden. PMID- 12902719 TI - [Helminthes Infections In The Small Intestine Of Stray Dogs In Ejungbu City, Kyunggi Do, Korea] AB - One hundred and two stray dogs in Ejungbu City of Kyunggi-Do, Korea were examined to reveal out the degree of natural helminthic infection of small intestine. Helminthes were collected at autopsy, by scraping the intestinal contents. The collected worms were classified by their morphological characteristics. Out of 102 examined, 72 dogs were infected with any helminthes. The common helminthes were Dipylidium caninum (47%), Ancylostoma caninum (26%), Toxascaris leonina (16%) and Toxocara canis (13%). Taenia pisiformis (9%), Echinostoma hortense (4%), E. cinetorchis (2%), Spirometra mansoni (2%) were also found. One dog was incidentally found to be infected with Clonorchis sinensis. The prevalence of Toxascaris leonina was relatively high in this study over Toxocara canis, and its significance was discussed. Dogs were firstly described as the reservoir hosts of Echinostoma hortense and E. cinetorchis in Korea. PMID- 12902720 TI - [An Epidemiological Study On Zoonoses In Korea] AB - Helminthic zoonoses are of significant importance to the public health and to the socioeconomic consequences especially in lost-meat, animal products and animal labor. For past several decades in Korea, it has been recognized that endoparasitic infections among domestic animals are extremely common and many of the helminth parasites of man have been acquired from, or are shared with domestic animals. However, there was no survey of a nationwide scale on the prevalence of animal helminthiasis; and control measures are also not satisfactory. Furthermore, because of the remarkable increase in the demand, number of domestic animals and pets is rapidly increasing in these days. Therefore, an extensive study on zoonotic helminths was carried out from July 1980 throughout August 1981 to understand the present status and parasite-host relationship from the epidemiological viewpoont. A total of 13,685 fecal specimens of cattles, sheeps, pigs, dogs and cats was collected from 26 localities in the country. The specimens were examined by formalin-ether centrifugal sedimentation technique. The results are summarized as follows: 1. Of 13,685 specimens examined, 9,808(71.7 %) were positive for helminth parasites. The positive rates were 99.0 % in cattles, 86.5 % on sheeps, 85.4 % in pigs, 35.4 % in dogs, 15.1 % in cats. 2. Single infection showed the highest rate (60.0 %), and double infection(27.5 %), triple(8.1 %), quadruple(2.8 %), quintuple(1.5 %) and hexad(0.1 %) were in decreasing order. 3. In the present study, 18 species zoonotic helminths were found: 11 nematodes, 6 trematodes and 1 cestode. 4. According to province, Cheju revealed the highest rate 84.8 %, then Kyungnam 77.5 %, Jeonnam 75.6 %, Jeonpuk 75.4 %, Kyonggi 75.2 %, Chungnam 71.7 %, Chungpuk 70.6 % and Kangwon 64.2 % were in decreasing order and Busan and Seoul showed 70.4 % and 58.1 %, respectively. 5. Rural area showed slightly higher rate (74.7 %) than urban area (68.5 %), and rates of mountainous area (70.4 %) and plain area (71.1 %) are very similar. 6. According to animal, helminths species and positive rates appeared in: Cattles: 6 nematodes and 3 trematodes; Strongyles 26.4 %, Neoascaris vitulorum 24.1 %, Trichuris ovis 12.0 %, Nematodirus sp. 6.3 %, Srongyloides papillosus 5.1 %, Capillaria bovis 0.02 %, Paramphistomum sp. 41.5 %, Fasciola hepatica 33.2 % and Eurytrema pancreaticum 29.7 %. Sheeps: 3 nematodes and 2 trematodes: Strongyles 33.0 %, S. papillosus 26.9 %, Nematodirus sp. 14.6 %, F. hepatica 31.7 % and E. pancreaticum 19.6 %. Pigs: 5 nematodes and 2 trematodes: Strongyles 43.1 %, Ascaris suum 29.0 %, Metastrongylus apri 20.3 %, Trichuris suis 14.0 %, Strongyloides ransomi 12.1 %, Clonorchis sinensis 1.4 % and Paragonimus westermani 0.9 %, Dogs: 4 nematodes and 3 trematodes and 1 cestode: Toxocara canis 14.4 %, Trichuris vulpis 9.5 %, Ancylostoma caninum 8.0 %, Toxascaris leonina 4.0 %, C. sinensis 2.4 %, Metagonimus yokogawai 1.3 %, P. westermani 0.8 % and Spirometra sp. 0.9 %, Cats: 2 nematodes, and 3 trematodes and 1 cestode: Toxocara cati 7.7 %, Ancylostoma tubaeforme 3.1 %, C. sinensis 1.9 %, P. westermani 1.4 %, M. yokogawai 1.2 % and Spirometra sp. 0.7 %. PMID- 12902721 TI - [A Human Case Of Thelaziasis In Korea] AB - The authors recovered a white thread-like living nematode in left conjunctival sac of a 33 years old male on May 29, 1981. The chief complaints were foreign body sensation and itching sensation of eye. The worm was a female measuring 12.1 mm in length and 0.171 mm in maximum width. It was identified as Thelazia callipaeda Railliet et Henry, 1910. And we recorded the patient as the 10th reported thelaziasis case in Korea. PMID- 12902722 TI - Splenic Paragonimiasis: A Case In A Dog. AB - A case of Paragonimus westermani cysts found deep inside the spleen is described. This spleen was from a dog that was experimentally infected via oral route with a hundred metacercariae and was killed 3 months after. At autopsy, beside the usual lesions due to Paragonimus in the lungs, the spleen was the site of multiple parasitic cysts. The involved spleen was slightly enlarged and showed an area of hemorrhagic dome on the capsular surface. There found a total of 8 adult worms in four well demarcated parasitic cysts containing a pair of worms each. All of them were found deep into the splenic parenchyme and were surrounded by a well formed connective tissue wall, thus resembling the features of the pulmonary paragonimiasis. The route of infection was assumed to be a direct transcapsular invasion during the phase of larval migration. PMID- 12902723 TI - [Studies on the mites(Order Acarina) in Korea: I. Suborder Sarcoptiformes] AB - Acarid mites were isolated from samples collected in Seoul during the period of October 1975 to August 1978. Samples consisted of stored foods such as rice, flour, sugar, dried fishes, dusts of houses and freight offices. The identified mites were classified into 12 species belonging to 12 genera, 5 families as following lists, of which 4 species (*) are newly recorded in Korea. Family Acaridae Ewing and Nesbitt, 1942 1. Tyrophagus putrescentiae (Schrank, 1781) 2. Acarus siro Linnaeus, 1758 *3. Aleuroglyphus ovatus Troupeau, 1879 4. Suidasia nesbitti Hughes, 1948 5. Rhizoglyphus echinopus (Fumouze and Robin, 1868) 6. Caloglyphus berlesei (Michael, 1903) 7. Lardoglyphus konoi (Sasa and Asanuma, 1951) Family Carpoglyphidae Oudemans, 1972 8. Carpoglyphus lactis Linnaeus, 1927 Family Glyphagidae Berlese, 1887 9. Glycyphagus destructor (Schrank, 1781) *10. Gohieria fusca (Oudemans, 1903) Family Chortoglyphidae Berlese, 1897 *11. Chortoglyphus arcuatus (Troupeau, 1879) Family Pyroglyphidae Cunliffe, 1958 *12. Euroglyphus maynei (Cooremant 1950) PMID- 12902724 TI - Comparative Efficacy Of Interval Mass Treatment On Ascaris Lumbricoides Infection In Korea. AB - To observe the effect of various interval mass treatment on Ascaris ova and worm positive rates and worm burden per population, 2-month, 4-month, 6-month and 12 month interval schemes were designed and applied. All of inhabitants in nine villages of Hwasung Gun, Kyunggi Do, Korea were treated from April 1977 to May 1980. Each village consisted of 100~140 people, however, there had been 20~40 drop-outs in every treatment. Pyrantel pamoate was used in dose of 10 mg/kg. Evaluation of schemes was made by examinations both for eggs and for expelled worms. The drop-outs in former treatment was included because they were few in munber. The reults obtained were summerized as follows: 1. The pre-treatment infection status of A. lumbricoides was not significantly different between Groups; 32.5-42.2% and 33.8-46.2% in egg and worm positive rates respectively. The mean worm burdens were in range, 1.6~4.2 per infected and 0.61~1.42 per population. 2. Twelve-month interval treatment was by no means meaningful to be adopted as a strategy of Ascaris control because egg and worm positive rates and worm burden were returned to pretreatment level. 3. The shorter the treatment interval was, the lower the egg/worm positive rates and worm burden were. By repeating biannual, triannual and six-times-in-a-year treatments, the indices of prevalence showed the tendency of further lowering during later 2 years of follow up. If the mass chemotherapy is adopted as a method of controlling ascariasis in a community, it should be repeated more than two times in a year to expect the gradual lowering of reinfection. PMID- 12902725 TI - Effect Of Two-Month Interval Mass Chemotherapy On The Reinfection Of Ascaris Lumbricoides In Korea. AB - Atrial to control reinfection cycle of Ascaris lumbricoides was made in a rural village in Korea by blocking the process of new egg production. Chemotherapy with pyrantel pamoate was repeated bimonthly in three hamlet groups, each consisted of 105~149 inhabitants of whole age group. In one hamlet (2.5 mg/kg dose) blanket mass treatment was repeated for 18 times from June 1977 to May 1980. In other two, 5.0 mg and 10.0 mg/kg dose groups, treatment was undertaken for 10 times from June 1977 until December 1987 and later reinfection patten was observed. Follow-up examination was made by whole-day stool collection for 2 days from each case after every treatment. The samples were examined both for eggs and worms, adult and immature young, of A. lumbricoides. The results obtained are summarized as follows: 1. The pre-treatment egg positive rate in Group A, B and C was 48.1, 45.3 an 37.8% respectively. 2. The lowest dose of pyrantel pamoate, 2.5 mg/kg was equally effective as higher doses in case of repeated use for reinfection control. 3. Among 4,311 pyrantel treatments, only 8 cases complained of side effects such as abdominal pain and diarrhea. 4. When the blanket mass chemotherapy was continued 18 times in Group A, all of the examined inhabitants were free from reinfection and egg production from 28th month (the 15th treatment) until the end of study period, for 8 months. 5. When the mass chemotherapy was stopped after 10 times, the egg positive rate returned gradually up to half of pre-treatment level within one year. 6. From the analysis of total 124 positive cases during the course of follow-up, it was revealed that the mass chemotherapy with 2-month interval was successful for the suppression of fertilized egg production. However, social factors such as infected immigrants or absentees were inevitably encountered and involved in the fertilized egg production. Summarizing the results, at least 28-30 months were needed to eradicate A. lumbricoides in the subjected area of Korea, by repeated blanket mass treatment bimonthly with a quarter dose of pyrantel pamoate. Furthermore, to lessen the social factors as a source of later reinfetion, the project area of control programme should be extended as wide as possible. PMID- 12902726 TI - Chronologic Growth Pattern Of Ascaris lumbrioides. AB - The chronologic growth patten of Ascaris lumbricoides was made by the maximum length of worms collected from inhabitants after mass chemotherapy with pyrantel pamoate by various interval; 2-month, 4-month, 6-month an 12-month. The results obtained were summarized as follows: 1. When the collected 497 worms were plotted on weight/length chart by treatment interval, there was always size limitation in each group, permitting the idea of maximal growth in a given period of infection. 2. The maximum lengths in each interval treatment, 2, 4, 6 and 12-month were; 12.5, 16.4, 19.2 and 22.8 cm in male and 14.2, 22.0, 26.2 and 30.8 cm in female respectively. 3. The maximum growth curves were expressed by the equations; Y=9.212 In(X+1)+0.025 in male and Y=11.953 In(X+1)+0.025 in female when X is age in month and Y is length in cm. 4. It was revealed that female grew more rapidly than male. The growth of length was rapid in initial 2~3 months and weigth gain was rapid in later than 2~3 months. The weight seems better than length as an indicator of aging. PMID- 12902727 TI - [Natural Transition Of Endemicity Of Malayan Flariasis In Inland Korea: Pattern Of Change In Microfilaria Rate Among Inhabitants Of Yongpung (Former Yongju) Area During The Period Of The Last Seven Years] AB - With a purpose to find out natural transition of endemicity of Malayan filariasis in inland Korea, a survey was conducted in June 1980 in Isan-Myeon of Yongpung Gun (former Yongju-Gun) where an epidemiological investigation had been carried out in 1973 without any control activities such as chemotherapy. Five sample villages were surveyed for microfilaremia by 20 microliter night blood examination among inhabitants and the results of the surveys conducted in 1973 and 1980 were compared to determine natural transition of the endemicity of malayan filariasis during the period of the last 7 years. 1. The current microfilaria rate among inhabitants in the 5 villages was 2.2 % on the average (male: 1.6 %, female: 2.8 %) from 370 persons examined. By village, the rates were 5.9 % (number of persons examined: 34) in Baranggol, 0 % (30) in Guitonggl, 4.2 % (72) in Alseonggol, 0 %(65) in Jangjagol and 1.8 % (169) in Saehae. 2. Extremely low microfilaria rate was noted in young age groups. By age group, no positive case was found in those age groups below 30~39 years except 10~14 age group in which 2 positives (4.4 %) were found. Two positives each were found in the respective age groups of 40~49(3.2 %), 50~59 (4.4 %) and over 60 (3.4 %). 3. In evaluation of the natural transition of the endemicity during the period of the last 7 years, the microfilaria rate turned out from 13.1 % in 1973 to 2.2 % in 1980. The difference in the microfilaria rate was 10.9 % and the natural reduction rate per year was 1.6 % on the average. 4. From the examination of 35 cases which had revealed microfilaremia 7 years ago, 85.7 %(30) of them were found to have converted to microfilaria negatives. On the other hand, from the 151 cases which had revealed no microfilaria in 1973, only 0.7 % (1) of them was found to have converted to microfilaria positive. 5. In the intensity of microfilaremia, the number of microfilaria/20 microliter blood per positive case was 11.0 in 1973 and 9.1 in 1980. The number of microfilaria/20 microliter per examinee was 1.4 in 1973 and 0.2 in 1980, thus reduced to 1/7 during the period of the 7 years. 6. The retarding endemicity of malayan filariasis in inland Korea was considered to be resulted in by the gradual increase of environmental factors in relation to ecology of vector mosquitoes, which adversely affect to the transmission of malayan filariasis. Followings are suggested to be the factors which control the transmission of the malayan filariasis in this area: 1) Inhabitants are the only natural final host of Brugia malayi infection in this area, 2) Gradual elevation of living standards of the inhabitants, 3) Gradual awakening of consciousness and behavior among inhabitants to protect themselves from mosquito biting using such as mosquito nets and insecticide sprays, 4) Preference of animal bait of vector mosquitoes, Anopheles sinensis, 5) Increase in number of domestic animals and fowls being raised in the village areas which play a major part of blood donors to vector mosquitoes, and 6) Relatively short (3~4 months) period of mosquito season in a year. PMID- 12902728 TI - [The Wormicidal Substances Of Fresh Water Fishes On Clonorchis Sinensis: III. Seasonal variation on the wormicidal substances of mucous substances of Carassius carassius] AB - The epidermal mucous substances of C. carassius which were extracted by ether were separated into various spots on the silica gel thin layer chromatography and separated into various fractions in the silica gel column chromatography, using petroleum ether (30 %) and chloroform(70 %) as a solvent. The wormicidal effects of the spots and the fractions on the cercaria, the liberating metacercaria and the adult of C. sinensis, were observed by month giving attention to ambient water temperature. 1. Among the four spots of the thin 1ayer chromatography of the ether extracts from C. carassius, the uppermost first spots (Rf. 0.924~0965) had a strong wormicidal effect on the cercaria in May, June, July, August and September, and the cercaria which came in contact with the materials was killed in 29~49 min., but in other months (Rf. 0.612~0.752) it was killed only after 50~115 min. 2. In column chromatography of ether extracts from C. carassius, the first fractions which had a wormicidal effect killed the metacercariae within 38~163 min. in May, June, July, August, and September, but it took about 120~458 min. to kill them in other months, meanwhile the wormicidal time of the fractions on the adults was 445~835 min. in May, June, July, August and September, but was 878~2,130 min. in other months. 3. The Rf. value of the uppermost first spot of the thin layer chromatography of ether extrart from the mucous substance of C. carassius which was reared in the aquarium controlled at 23 degrees C was 0.79, and its first fraction of column chromatography killed the metacercaria and the adult of C. sinensis within 315, and 1,850 min. respectively. As summarized above, the wormicidal materials of the epidermal mucous substances of C. carassius had no relation to ambient water temperature, but the secretion period of this wormicidal materials was the same as the liberating period of the cercariae from Parafossarulus manchouricus. PMID- 12902729 TI - [In Vitro Activities Of Various Drugs Against Adult Worms Of Clonorchis Sinensis] AB - The present study was undertaken to evaluate in vitro activities of following drugs against adult worms of Clororchis sinensis: i.e., stibnal, chloroquine diphosphate, Hetol (1,4-bis-trichloromethylbenzol), dehydroemetine "Roche" (Ro 1 9334), niridazole (Ambilhar), bisbendazole, gentian violet, dithiazanine iodide, hexachlorophene, bithionol, niclofolan (Bilevon, Bayer 9015) and praziquantel (Embay 8440, Biltricide). After isolation from bile ducts of experimentally infected rabbits (3 months infection), the parasites were rinsed in sterilized Tyrode's solution and incubated for 24 hours at 37 degrees C in TC 199 medium with Earle's balanced salt solution containing 0, 0.01, 0.1, 1.0, 10 and 100 microgram/ml concentrations of each above drugs. The activities of each concentrations with each drugs were evaluated by the motility of the worms in culture tubes observing at 15, 30, 60 minutes and 2, 4, 6, 12, 18 and 24 hours by stereomicroscope. In the results, no effect was found at the adult worms of C. sinensis in all concentrations with stibnal, chloroquine diphosphate, Hetol, dehydroemetine, niridazole and bisbendazole. However, moderate activities were observed in the concentrations of 1.0 and 0.1 microgram/ml of gentian violet, dithiazanine iodide, hexachlorophene and bithionol. The highest activities were observed in all concentrations of niclofolan and praziquantel. PMID- 12902730 TI - [Morphological Observations On Clonorchis Sinensis Obtained From Rats Treated With Praziquantel] AB - The present study was undertaken to observe the morphological changes of Clonorchis sinensis obtained from experimentally infected rats treated with praziquantel (Biltricide) which is a new anthelmintic agent with high activity against a broad spectrum of cestode and trematode species. For this study, the rats were infected experimentally with about 50 metacercariae of C. sinensis and were given praziquantel a single dose of 600 mg per kg of body weight at 5 weeks after infection. The Clonorchis worms were isolated from the bile ducts of the rats which were autopsied on the 4th day after treatment. After isolation the parasites were observed their shape and motility in the medium of 37 degrees C physiological saline solution, and then the fine structure of the tegument of C. sinensis was studied by means of light, scanning and transmission electron microscope. The findings of the observation were compared with those of untreated parasites. The results are as follows: 1. All the isolated worms moved actively in the medium of physiological saline solution (37 degrees C). A majority of the parasites obtained from the treated rats showed a large balloon shaped structure on the surface between oral and ventral suckers. But such structure has never seen in the parasites obtained from control rats. 2. By the scanning electron microscopic observation, the regular pattern of the tegumental ridges was significantly changed on the outer surfaces of parasites obtained from the treated rats as compared with those of the control rats. 3. By the transmission electron microscopic observation, the numerous mitochondriae in the syncytial tegumental layer of the treated parasites appeared to be degenerated and formed small vacuoles, and the tegumental ridges were also degenerated and showed somewhat flattened. 4. In the parasites obtained from the treated rats, a different size of vacuoles interspersed in the distal part of the syncytial tegument and also in subepithelial region of the urinary bladder. These vacuoles are fused each other and lead to the disruption of the apical region of syncytial tegument along the basement layer. Finally the basement layer was dislocated. So that the tegumental layer appeared as a large balloon. PMID- 12902731 TI - Epidemiological studies of Clonorchis sinensis in vicinity of river Taewha, Kyungnam province, Korea. AB - In order to determine the epidemiological patten of the liver fluke, Clonorchis sinensis, the locality where the snail, Parafossarulus manchouricus, is abundant, the infestation rates for the cercarial and metacercarial larvae of digenetic trematodes in the snail and fish hosts, and the prevalence of Clonorchis sinensis among the residents in the vicinity of river Taewha, Kyungnam Province, Korea were studied from March 1979 to September 1980. Four snail habitats were found in the river Taewha. The population density of the snail per square meter of the habits ranged from 10 to 500, with an average of 150. Among the 4 snail habitats, three, Baek-cheon, Sa-yeon and Taewha had snails infected with the cercaria of Clonorchis sinensis, and the proportion of infested snails in the three habitats was very low, the average being 0.59 per thousand. Four species of fresh-water fish, the bullhead, Coreobagrus brevicorpus, the Korean shiner, Gnathopogon atromaculatus, the flat bitterling, Paracheilognathus rhombea, and the striped shiner, Pungtungia herzi among ten species of fish examined harbored the metacercaria of Clonorchis sinensis. The intensity of infestation with the cyst of Clonorchis sinensis in the four species of fish was low. The approximate number of the cysts per gram of flesh was between 3.5 in Paracheilognathus rhombea and 21.5 in Gnathopogon atromaculatus. Seven kinds of laval tremathodes, Clonorchis sinensis, Cyathocotyle orientalis, Echinochasmus species, Exorchis oviformis, Metacercaria hasegawai, Metagonimus yokogawai and Metorchis orientalis were found from ten species of fish examined. In general, the infestation rates for the encysted larvae of digenetic trematodes varied from fish to fish. The infection rate for Clonorchis sinensis among the residents in the vicinity of river Taewha was relatively high, 22.2 % by the formalin-ether sedimentation technique, and the difference in the rate of infection between males and females found to be siginificant(t > 2). Of these, the rate was 12.4 per cent in 483 elementary school children and 17.7 % in 761 middle and high school students. The results obtained in the present study indicate that endemic foci of Clonorchis sinensis exist in the river Taewha and the prevalence of the liver fluke among the residents is still high. PMID- 12902732 TI - [Study on the Metagonimus sp. in Gum river basin, Chungchung-nam Do, Korea] AB - This study was undertaken to elucidate the status of Metagonimus species infection and to demonstrate the first and the second intermediate hosts of the fluke at the several localities along the Gum river, Chungchung-nam Do, Korea from March 1979 to October 1980. The results are summarized as follows: 1. Total 790 stool specimens of inhabitants from 6 localities along Gum river were examined by the combination of cellophane thick smear technique and formalin ether concentration technique. The positive rate of intestinal helminths was 80.3 %, and the egg positive rate of Metagonimus sp. was 21 % from a total of 790 specimens examined. In the age bracket of 20~49, the positive rates of Metagonimus sp. were 31.1~36.5 %, and the males (26.8 %) were infected three times higher than females (8.9 %). By the area of the river, Gumsan which is located upper stream showed the highest positive rate (40.5 %) of the fluke, whereas the rates decreased gradually toward upper part of river (Muju) or the descending lower area of the river. The most lower area of river (Gangkyeong and Nonsan), no positive Metagonimus infection case was found. 2. The infection rate of Clonorchis sinensis in 790 was 37.3 %, and the status of mixed infection of Metagonimus with C. sinensis was found 127 cases (76 %) out of 166 cases with Metagonimus infection. 3. The result of measurement of ova from 166 cases of Metagonimus infection, large ova were found in 41.1 %, small size ova in 31.9 % and the mixed type in 27.1 %, respectively. The adult worms were recovered from the 6 cases after treatment with niclosamide, and the measurements of the worms showed similar morphological characteristics of the Metagonimus yokogawai. 5. Twenty-two species in 5 families of fresh water fish were caught. Out of 22 species of fish, 18 species were found infected with metacercaria of Metagonimus. In order to observe the distribution of metacercariae in the body and the rate of infection of the individual host, a total of 94 Pseudogobio esocinus were examined. Total 84 (89.4 %) out of 94 fish were infected with the metacercariae and the rates of distribution of the metacercaria were observed in the following order: on scale; 93.4 %, in gill; 3.2 %, head part; 2.9 %, but a few metacercaria was found in muscle and visceral organs. 6. The development of the metacercariae which were isolated from the scale of Pseudogobio esocinus were experimentally traced in the albino rat host from one day to 30 days after infection. At the 6th day, several eggs were visible in the uterine tubule of the some worms, on the 10th day, worms became 326.0 x 208.0 micrometer in size, however the bodies of worms grew no more until the 30th day after infection. The oral sucker was larger in size than the ventral sucker during early stage of the worms, but the 7th day after infection, it reversed. The ventral sucker situated on the ventral right side of the body. Right side of testis laid somewhat toward, and uterus and vetellaria extended to the posterior margin of the right testis. 7. Snails(Koreanomelania nodifila var. ovulum) were collected in the river, Buri Myeon, Gumsan-Gun, and found the natural emerging ophthalmo-pleurophocercus cercaria. From the above result, the prevalence of Metagonimus species infection showed relatively higher rates from the inhabitants along the middle to upper localities of the river. Even though no sweetfish was caught in the surveyed areas, Gum river basin is the one of the new endemic foci of metagonimiasis in Korea. PMID- 12902733 TI - Epidemiological study of Paragonimus westermani in Wiseong County, Kyungpook Province, Korea. AB - This survey was undertaken to determine the recent prevalence of lung fluke, Paragonimus westermani, in the three localities of Wiseong county, Kyungpook Province, Korea and to compare the prevalence among the residents in the same areas between 1924 and 1980. The three localities, Ku-seong, Sam-chun, and Um-ji villages (dong) have been known as the endemic areas of Paragonimus westermani. The surveys were made on the general population. They were examined using intradermal test with Paragonimus westermani antigen, and the diagnosis of paragonimiasis was made depending on the positive intradermal reaction. The sputum examination could not be performed beacause the residents were inaccessible. The prevalence of Paragonimus westermani among the residents in the three communities varied from 1.4 % to 7.9 per cent, with an average of 4.0 per cent. Whereas, the average reported by Ichinomiya in 1924 was 4.7 %. There are no significant differences in the sex and age specific rates of infection between 1924 and 1980. With the abundant density of Semisulcospira snails and crayfish in Sam-chun stream and the high rate of infection with Paragonimus westermani among the residents in the village, it is found that Sam-chun village, one of the three localitied in Wiseong county, where Ichinomiya (1924) surveyed, was still the endemic focus of Paragonimus westermani. PMID- 12902734 TI - [An Epidemiological Note On The Taeniasis In Korea] AB - The present study was undertaken to evaluate the prevalence rate of Taenia spp. infection and distribution of taeniasis caused by T. solium among Koreans in several localities in Korea during the period from 1977 to 1980. A total of 1,946 stool specimens collected from students and inhabitants of Chungcheong Bug Do and Gyeongsang Nam Do were examined by cellophane thick smear method. Among them, 40 persons were detected as Taenia egg positive cases. The overall positive rate of Taenia spp. shows 2.1 %. The prevalence rate of male (2.3 %) is relatively higher than that of female (1.4 %). In order to observe the distribution of Taenia solium infection, the whole worms or a part of proglottids of Taenia spp. were collected from the stools of egg positive cases by normal defecation or anthelmintic treatment. For the species identification, expelled proglottids were examined microscopically by the number of branches of the uterus, presence of a vaginal sphincter or the accessory ovarian lobe etc. Among 199 egg detected cases in this study 59 (39.1 %) out of 151 cases in Seoul, 12(40.0 %) of 30 cases in Gyeongsang Nam Do and 2 (25.0 %) from 8 taeniasis cases in Cheju Do were infected with T. solium. But none of T. solium infection was found from 10 egg positive cases in Chungcheong Bug Do. As a whole, the composition of species shows 36.7 per cent of T. solium infection and 55.8 % of T. saginata infection, and in 7.5 per cent the species were not identified. PMID- 12902735 TI - Demonstration Of Entamoeba Histolytica Cyst From Urban And Rural School Children. AB - School children between 6 to 12 years of age were studied for the presence of Entamoeba histolytica cysts in a period of 6 months, form March to August 1980. Fecal specimens collected form urban and rural children were examined first by the Lugol stained technique and then by the Faust's iron alum hematoxylin method. A total of 731 specimens were examined. Of these 26.9 per cent, 197 specimens, harbored the cysts. The demonstration rate for the cysts by age group varied from 19.6 to 43.5 per cent. The demonstration rate in boys was 24.4 per cent and in girls 29.8 per cent. Similarily, the rate in urban and rural area was 32.4 and 24.1 per cent, respectively. It was found in the present study that there were no significant differences in the age and sex rates of demonstration between urban and rural children but there was a predominance of the small race of Entamoeba histolytica over the large race. PMID- 12902736 TI - Prevalence of Trichomonas vaginalis in Korean military Personnel. AB - A study of Trichomonas vaginalis in Korean military personnel, based on the discovery of motile trophozoites by prostate culture and urine wet smear methods, has been carried out during the period from May, 1975 to October, 1977. In the Army General Hospital clinics, specimens of prostatec secretion and urine were provided by the in- and out-patients without any noticeable genito-urinary symptoms. In a total of 977 specimens 33 cases were revealed positive for Trichomonas vaginalis. Of these, 23 cases were positive in both prostate culture and urine wet smear methods, 8 positive in the culture but negative in the wet smear, and 2 positive only in the wet smear. In comparing the demonstration rates for trichomonads by culture with those by smears, the former was more likely to give a positive result than the latter. The pattern of the incidence rate by age group was appreciably varied; 2.1 per cent in the 20-24 age group and 5.1 per cent from 25 to 29 and subsequently increased and reached a maximum of 5.7 per cent in the 40 and over age group. It was found that Trichomonas vaginalis in Korean military personnel is one of the important diseases with a high indicence, and that the eradication of this disease is possible with extensive public health education and the administration of specific therapeutic agents to patients with trichomoniasis. PMID- 12902737 TI - Fine Structure Of Cysticercus Celluosae From Human Brain. AB - A transmission electron microscopic study was conducted in order to know basic tegumental structure of Cysticercus cellulosae which was obtained from the ventricles of human brain. In general, the electron microscopic appearance of the tegument resembled that of the body wall of adult and larval forms of other cestodes. The tegument both of the parenchymatous portion (scolex and spiral canal) and bladder portion consisted of such components, i.e., an outer vesicular layer with microtriches, and an inner fibrous layer. The subtegumental tissue below the fibrous layer was filled with muscle bundles and tegumental cells. The surface of the larva was covered by the microtriches of two distictly different types. The characteristic pyramid-shaped "tetrahedral" form was observed on the surface of the scolex portion, whereas the elongated, slender "filamentous" microtriches were distributed on the regions of the spiral canal and bladder portion. The tegumental and subtegumental tissues varied in thickenss from one region to the next. The possibility of application in making differential diagnosis from other larval cestodes and possible fuction of this larval tegument were discussed. PMID- 12902738 TI - Histopathologic Study On Human Sparganosis. AB - Based on 16 cases of human sparganosis, a histopathological study was made. There was a striking similarity among histological features of sparganosis involving different tissues. The histological change of the affected tissues was characterized by a necrotizing and granulomatous inflammation with or without worm parasite in the lesions. There was also a remarkable polymorphonuclear leukocytic mobilization, predominantly of eosinophils, plasma cells and lymphocytes in and near the lesions. Tunnel formation lined by palisading histiocytes was another charateristic feature of the host tissue reaction. These findings were quite distinguishable from those of cysticercosis which were more localized and self-limited. Several features that were prominent in section slides of sparganum worm parasite were also noted. Laminated calcospherules found in the cytoplasm of the proliferating macrophages and giant cells were of diagnostic value of sparganosis in the absence of the worm, particularly when these were accompanied with tunnel-like lesion in the host tissue. PMID- 12902739 TI - Anthelmintic effect of amidental (Bay d 8815) against Ancylostoma duodenale infection. AB - A new anthelmintic, amidantel(Bay d 8815), an acetylated p-amino-phenyl acetamidine was tried in 140 patients with Ancylostoma duodenale and other helminth infections. In the first trial, each 16 cases in 64 patients with A. duodenale were treated with 3.0, 6.0 and 9.0 or 10.0 mg/kg body weight of amidantel including placebo control. Another 76 patients infected with hookworms and other helminths were treated with 5.0, 6.0 and 8.0 mg/kg body weight of amidantel in the second trial. In order to assess the efficacy and safety of the drug, follow-up examination by repeated and replicated examinations over three consecutive days were performed at 14 to 16 days and 28 to 30 days after treatment, And complete laboratory studies including ECG were carried out before and one day after the medication. In the results, it was confirmed that amidantel is very effective against A. duodenale as well as Ascaris lumbricoides. With regard to dosage, a single dose of 6.0 mg/kg body weight of amidantel was found to be the most effective and well tolerated than the other dosages employed. In a single dose of 6.0 mg/kg body weight the cure rates were 93.8 and 96.6 per cent for A. duodenale infection and 90.9 and 93.1 per cent for ascariasis in the first and second trials respectivley. Relatively significant activity was also observed against Necator americanus at the dosages employed, however it was not superior to other drugs currently use. No significant activity was noted against Trichuris trichiura. Side effects including headache, nausea, dizziness and abdominal discomfort were usually mild and transient. No significant changes attributable to therapy were observed in hematology, blood biochemistry and urinalysis as well as ECG. PMID- 12902740 TI - Chemotherapeutic control of Enterobius vermicularis infection in orphanages. AB - To observe the effect of egg-free state in an institute on the endemicity of Enterobius vermicularis, the theoretical condition was made by repeated chemotherapy. Mebendazole or pyrantel pamoate were administered to all orphans and their staffs concomitantly for four times, each 20-day interval. In two low endemic orphanages, in which mean egg positive rates were 18 % and 39 % respectively, the pinworm infection was controlled up to 6~7 months period. And other two highly endemic orphanages, with 70% and 55% of egg positive rates, were controlled up to 90th day after the first treatment. After then up to 7 months, the reinfection occurred, but the endemicity was brought down to about 20 % of egg positivity. PMID- 12902741 TI - Selection Criteria Of Subjects For Drug Evaluation In Enterobiasis. AB - When an antipinworm drug was evaluated in human being, the results are still variable according to authors. Among the various factors involved in these variable results, the selection of subjects must be one of them. In the past, in evaluating a drug, the subjects were simply anal swab positive cases in a population. By that kind of selection method, especially in low endemic population, anal swab positives spontaneously turned to egg negatives at random. Moreover, the time, frequency of negative conversion cannot be predicted. By the results, the cure rates obtained from such ill-defined subjects are variable according to the subjected population. Another problem is that evaluation should represent the efficacy in heavily infected cases. To reinforce and overcome such shortcomings, the evaluator shoud select the cases wha are infected with whole developmental stages of worm. And the evaluator should have a method selecting such ideal cases only with anal swabs. To find out such method the results of anal swabs of long-term follow-up without any antipinworm treatment were reevaluated. In 41 children from 6 orphanages of different endemicity, swabs were taken for 8 or 21 times during 36 or 91 days. The positive rate of follow-up swabs was compared with that of already obtained in 4 preliminary swabs. And we collected other data on the combination of 4 anal swab results in different endemicity. The results are as follows: 1. The positive rate of follow-up swabs was significantly different by the consecutiveness of egg positivity in preliminary swabs as well as the endemicity of subjected population. The consecutively positive cases in higher endemicity over 60% showed the highest, and the least variable positive rate in follow-up swabs. 2. The observed frequency of consecutively positive/consecutively negative combination was always higher than the expected one obtained by binomial distribution equation. This indicates that individual hygiene and previous infection are important factors in determining the actual occurrence of subsequent reinfection. Summarizing these results, the subjects for drug evaluation should be those of consecutively positive cases in population over 60% endemicity. These subjects have the lowest chance of spontaneous negative conversion at random period of follow-up swabs. Eligible cases for such criteria would be sufficiently secured in highly endemic population. PMID- 12902743 TI - [Experimental study on the therapeutic effect of praziquente1(Embay 8440) in rats experimentally infected with Clonorchis sinensis] AB - Experimentally infected rats with 50 metacercariae of Clonorchis sinensis each were treated orally with different dosages of praziquantel. About 100 rats were divided into 8 groups in each developmental week after infection (from the first to 8th week after infection) and 24 rats served as untreated controls. In each developmental week after infection, about 15 rats divided into 4groups including control group(3 to 5 rats) were used to determine for the curative activity of praziquantel against C. sinensis. The experimentally infected rats in each developmental weeks after infection received praziquantel 1 x 100 mg/kg x 3 days, 3 x 100 mg/kg x 1 day and 3 x 100 mg/kg x 2 days. The effects of the drug were evaluated by the worm recovery rates, morphological changes of the worms and the number of eggs per gram of feces (EPG) after treatment. The feces of individual rats were collected once in a week at one week before and 4 weeks after treatment, and the number of Clonorchis eggs were examined by Stoll's egg counting method. The necropsy was performed on 4 weeks after treatment. The liver and the bile ducts were examined immediately after the dissection, and all of the worms recovered from the liver immersed in warm saline solution in a small petri dish in order to observe the motility of individual worms. After that, all worms obtained from the treated and untreated control rats were fixed in 10 percent formalin solution. The whole mount specimens were prepared by Semichon's acetocarmine stain for the study of morphlogical changes. In the treatment with dosage of 1 X 100 mg/kg for 3 consecutive days, relatively low worm recovery rates were shown in the early stages of infection than those of control groups. However there was no differences of the worm recovery rates between the later stages of infection (7 to 8 weeks after infection) and control groups. In the early stages of infection, the worms recovered in the bile ducts of each treated rats showed morphologically intact as normal control groups, however in the later stages of infection the worms showed damage of their reproductive organs such as testes, seminal receptacle and ovary etc. In the treatment with dosage of 3 X100 mg/kg X 1 day, the recovery rates of the worms in all developmental weeks after infection were considerably low and some growth inhibition of the worms were also noted. The worms recovered from the bile ducts in all developmental stages were seriously damaged in their reproductive organs, such as testes, seminal receptacle, ovary, Mehlis' gland, uterus and vitelline glands etc. The hyaline degenerations were also observed on the parenchymal tissues in the dead worms. The mean EPG values in this group were markedly reduced at the first 2 weeks after treatment. However the low values of EPG were generally noted at the 3rd to 4th week after treatment. On the other hand, in the treatment with dosage of 3 X 100 mg/kg X 2 days, no more the living worms was recovered in all developmental stages of Clonorchis. The dead worms recovered from the bile ducts were so seriously damaged that the morphological changes could not be observed. The mean EPG values in this group were markedly reduced during the first 2 weeks after treatment. At the 3rd week after treatment EPG value became negative in the most of all treated groups. No more the Clonorchis eggs were detected at the 4th week after infection, but only a very small numbers of eggs were found by the concentration method. From the above results, it is obvious that praziquantel exhibits a high degree of activity against Clonorchis sinensis in the rats, with no apparent differences in the efficacy against each developmental stage. PMID- 12902742 TI - Seasonal variation of snail population of Parafossarulus manchouricus and larval trematode infection in river Kumho, Kyungpook province, Korea. AB - Studies were conducted to determine the ecology of the snail, Parafossarulus manchouricus, the seasonal variation of the snail population, and the infestation rates of these snails with the cercariae of digenetic trematodes, in the snail habitat of river Kumho, in Dongchon of Taegu city, in Kyungpook Province, Korea, from 1975 to 1979. The earliest time the snails were found was early in April every year. At that time the water temperature ranged from 13 to 14 degrees C. The population density of snails ranged from 2 to 8 per square meter. The highest population density of snails was encountered in late June and the water temperature was between 24 and 26 degrees C. The snails disappeared in early and mid-November, when the water temperature dropped to 10~13 degrees C. The average snail population in June was 115.9 per square meter in 1975 but in 1976 dropped abruptly to 30.5 and remained at approximately the same level in 1977. It then decreased at considerably lower rates, with the average of 18.7 in 1978 and 14.5 in 1979. The snails collected in the habitat were examined for the presence of cercariae of digenetic trematodes. Of these, four species, Clonorchis sinensis, Cyathocotyle orientalis, Exorchis oviformis, and Loxogenes liberum were found. The most frequently liberated cercaria was L. liberum 43.3 per cent, followed by E. oviformis 6.2 per cent and C. orientalis 1.5 per cent. The least frequently liberated was C. sinensis 0.1 per cent. In the monthly liberation rates for larval trematodes, the higher rates were observed in June, July and August every year, and the cercarial rates for Clonorchis sinensis, Cyathocotyle orientails and Exorchis oviformis demonstrate a fairly consistently fluctuating downward trend over the five-year period, except for Loxogenes liberum. Summarizing the results, this study indicated that the population density of snails in the habitat and the infection of the snail with the cercariae of digenetic trematodes has decreased over the five-year period from 1975 to 1979. PMID- 12902744 TI - [A Study On The Fine Structure Of Clonorchis Sinensis, A Liver Fluke: II. The Alimentary Tract And The Excretory System] AB - A morphological study on the ultrastructures of the alimentary tract and the excretory system of Clonorchis sinensis was conducted. The liver flukes were collected from rabbit liver six months after the experimental infection The worms were washed with 0.85% saline solution and immediately moved to cold 2.5% glutaraldehyde in 0.1 M phosphate buffer pH 7.4. The materials were dissected and fixed for two hours. The blocks were post-fixed in 1% osmium tetroxide. The blocks were embedded in Epon 812. Ultra thin sections were cut with Sovall MT-2 ultramicrotome and stained with uranyl acetate and lead citrate. Sections were then observed with Hitachi HS-7S electron microscope. The following results were obtained in a series of observations. The walls of oral cavity and esophagus comprised tegumental syncytium, basement membrane, loose connective tissue, muscular layer and parenchymal cells. The apical surface and the base of the syncytium were covered with a protoplasmic membrane for each forming numerous invaginations. Granular endoplasmic reticulum was developed in the epithelium of the oesophagus. The gastrodermis of Clonorchis sinensis comprised two types of cells in general. The first cell type was numerous one forming a single continuous layer of epithelial cells. Each of the cells had outfolded cytoplasm into the caecal lumen and lamellae along the cell surface. Among the above epithelial cells, no considerable differences in structure reflecting their functional states were identified. The second cell type was less differentiated in nature and lay within the gastrodermis above the basement membrane but not in contact with the caecal lumen, being overlapped by neighboring gastrodermal cells of the type described above. At this portion the gastrodermis seemed to be a pseudostratified epithelium. There were well-developed lamellae along the surface of epithelia of all canals or duct concerning evacuation. The excretory pore was 7.5 micrometer in diameter and dorso-terminally opended. The epithelium of the excretory pore, a syncytial layer, contained many microtubules unlike the other part of tegumental layer of this worm. The epithelium thickness of the excretory pore was very irregular(1.3-5.5 micrometer). PMID- 12902745 TI - [A Study On The Fine Structure Of Clonorchis Sinensis, A Liver Fluke: III.The Prostate Gland] AB - A study on the ultrastructures of the prostate gland of Clonorchis sinensis was conducted. The presence of the prostate gland in this fluke has not been known up to present time. Authors observed the ejaculatory duct epithelium including its surrounding parenchyma and found the prostate gland with the help of the electron microscope. The prostate gland was consisted of numerous unicellular glands grouped around the ejaculatory duct. The individual cell was lobulated, tapering in the direction of the ejaculatory duct. The secreting ducts of the gland penetrated into the ejaculatory duct through the muscular layer and the basement membrane, and finally opened to the lumen of the ejaculatory duct. The secreting duct had single layered microtubules along the inner wall of the duct. The secretory bodies produced by the prostate gland seemed to be moved to the lumen of the ejaculatory duct through the secreting duct. The prostate gland of this fluke was less developed than that of Fasciola hepatica but the basic structures were quite similar. There were well-developed lamellae in the epithelia of all ducts concerning passage of spermatozoa from the testes to the male genital opening. PMID- 12902746 TI - [The Wormicidal Substances Of Fresh Water Fishes On Clonorchis Sinensis: II. Preliminary Research On The Wormicidal Substances From Mucous Substances Of Various Fresh Water Fishes] AB - The present work which was investigated in July 1979, was to observe the wormicidal effects of the external mucous substances of 9 species of fresh water fishes (Cyprinus carpio, Parasilurus asotus, Anguilla japonica, Ophicephalus argus, Carassius carassius; golden crusian carp, Misgurnus anguillicaudatus, Zacco platypus, Pseudorasbora parva and Carassius carassius; crusian carp) on cercaria, liberated metacercaria and adult of Clonorchis sinensis. The mucous substances extracted by ether from the above 9 species of fishes were separated into many spots on the silica gel thin layer chromatography, and divided into many fractions in the silica gel column chromatography which used petroleum ether (30%) and chloroform (70%) as a solvent. 1. On the silica gel thin layer chromatography of ether extracts from 9 species of fresh water fishes, each of the Rf. values which had wormicidal effect on the cercaria of C. sinensis were different from others; Rf. value of C. carassius (crusian carp) was 0.937, 0.709 in O. argus, 0.612 in A. japonica, 0.576 in P. asotus, 0.451 in C. carpio, 0.701 in Z. platypus, 0.385 in C. carassius(golden crusian carp) and 0.15 in P. parva. Time for wormicide was different from each other, too. It took 14 min. in a case of C. carassius(crusian carp), 25 min. in Z. platypus, 26 min. in C. carassius(golden crusian carp), 28 min. in C. carpio, 30 min. in P. asotus, 35 min. in O. argus, 40 min. P. parva and 180 min. in A. japonica. But any of the spots of M. anguillicaudatus did not show wormicidal effect on the cercaria within 10 hours. 2. In the silica gel column chromatography of ether extracts from 9 species of fresh water fishes, the fractions which had wormicidal effect on the metacercaria of C. sienesis were different from each other; in a case of C. carassius(crusian carp), the first fraction only had wormicidal effect, the 2nd in P. asotus, the 3rd in O. argus, the 4th in C. carassius(golden crusian carp), the 2nd in C. carpio and the 4th in P. parva. Time for wormicide was 40 min. in C. carassius(crusian carp), 52 min. in P. asotus, 74 min. in C. carassius(golden crusian carp), 92 min. in O. argus, 95 min. in C. carpio and 140 min. in P. parva, but any of the fractions of A. japonicas, M. anguillicaudatus and Z. platypus did not show wormicidal effect within 10 hours. 3. The fractions which had wormicidal effect on the metacercaria of C. sinensis had also the same effect on the adult, but it took longer time to kill them. It took 3 hours in a case of C. carassius(crusian carp), 4 hrs. in C. carassius(golden crusian carp), 5 hrs. in P. asotus, 6 hrs. and 6 min. in O. argus, 6 hrs. and 10 min. in C. carpio and half and 6 hrs. in P. parva. But any of the fractions of A. japonica, M. anguillicaudatus and Z. platypus did not show wormicidal effect within 24 hrs. PMID- 12902747 TI - [A Note On Intermediate Hosts Of Paragonimus In Discrict Of Seoul] AB - Recent results of mass stool examination for parasitic infections revealed 0.01~0.0001 percent of Paragonimus egg positive rate in Seoul school-children. This survey was carried out to understand the infection status of intermediate hosts and their relations to human paragonimiasis in Seoul. Twelve mountainous streams in the outskirts of Seoul were surveyed. From all streams, Semisulcospira libertina, the snail intermediate host was not detected. From 7 streams 132 crayfishes, Cambaroides similis, the second intermediate host were collected. From them no metacercaria was found. The results indicated that crayfish in district of Seoul is no more an infection source of paragonimiasis. Possible factors related with the ecology of intermediate hosts were briefly discussed from the epidemiological view point. PMID- 12902748 TI - [Prevalence Of Capillaria Hepatica Among House Rat In Seoul] AB - During May-July, 1978 a survey on the prevalence of C. hepatica among house rat(Rattus norvegicus Erxl.) and wild mice (Mus musailus molossinus and Microtus mandarinus) in two districts in Seoul was carried out. Approximately 38% of 1,000 house rats examined were infected. Rat population as an established group in Ahyundong, neighbouring the city, revealed 39.2% of infection among 660 house rats, and that as newly formed group in Shinrimdong, a developing residential quarter on the southern outskirts of Seoul showed 35.9% among 340 rats. Infection rate of females and males was 38.9% and 37.2% respectively. Differences of infection rates by locality and sex were not statistically significant. It was very interesting that none of 64 wild mice were infected with this parasite in both districts. Some probable factors which are related on C. hepatica infection among rats were discussed briefly. PMID- 12902749 TI - Egg Discharging Patterns Of Ascaris Lumbricoides In Low Worm Burden Cases. AB - In Ascaris lumbricoides infection, the faecal examination, undertaking for ova detection, is not always diagnostic. It is just because some of the infected cases has Ascaris worm(s) which do not produce eggs. In the present study, the authors attempted to analyse quantitatively the egg discharging patterns in Ascaris infected cases with low worm burden. The following results were obtained: 1. In 1,861 Ascaris worms collected from 853 cases, the sex ratio was 1: 0.74 (female: male). Sex combinations in each burden of case were always fitted with theoretical values from the binomial distribution; (m+f)(n)= (0.426+0.574)(n). 2. In each worm burden, their sex combination indicated different egg discharging patterns; false negative cases, unfertilized ova passers and fertilized ova passers. When the relative frequency of the above three egg discharging patterns was plotted to worm burden per case, a definite relationship was found. The cases with six or more worms have nil probability to be false negative case or unfertilized ova passer. 3. Out of 853 cases, we found 129 false negative cases. The collected worms from 125 cases were morphologically analysed. It was found that 52% of them were infected with only male worm(s) and 24% were infected with young female worm(s). And in 8.8%, old female(s) with empty uterus were infected. The cause of 15.2% was remained unexplained, even though the collected worms were scrutinized. PMID- 12902750 TI - Frequency Distribution Of Ascaris Lumbricoides In Rural Koreans With Special Reference On The Effect Of Changing Endemicity. AB - This study was undertaken to figure out the basic patterns of the frequency distribution of Ascaris lumbricoides in rural Koreans and to find out the effects of changing endemicity on the frequency distribution pattern. Total 853 cases in 6 areas were surveyed from November 1975 to February 1978. The mean ova positive rate was 33.1% and in the range from 23.9 to 66.7% by the surveyed unit. From the analyses of the collected data, the following results were obtained. 1. The overdispersion pattern of worm burden per case was mathematically fitted both to the theoretical values of the negative binomial and Poly-Eggenberger distributions. However, when "0" and lower burden cases were considered, the better fitness was observed in the negative binomial than in Polya-Eggenberger distribution. 2. Although the endemicity of Ascaris is changed, the relative frequency of the lower burden cases is rather constant. 3. The false negative rate of stool examination and unfertilized ova passers in the population were not changed significantly according to the endemicity. And the rate of unfertilized ova passers in infected cases was increased when the endemicity was lowered. From the above results, it is concluded that the overdispersed frequency distribution pattern of Ascaris in an endiemic area is changed by the relatively rapid decrease of heavy burden cases and increase of non-infected cases according to the lowered endemicity due to the impact of the control. PMID- 12902751 TI - [A Mathematical Approach To The Mode Of Transmission Of Clonorchiasis In The Inhabitants Of Nak-Dong And Han River Basin] AB - To understand the mode of transmission in clonorchiasis, a survey was made in Kim hae Goon, South Kyong-sang Do (=Province). The mathematical analysis of the age prevalence was done on the egg positive rates. And another analysis for the comparison was also made to the cited data from two areas, North Kyong-sang Do and Ko-yang Goon, Kyong-gi Do. Some catalytic models of H. Muench (1959) were applied to the observed age prevalence. Because the both parameters, such as force of infection(a) and loss of positivity(b) were considered to be constant for a long period in the surveyed area, the two stage catalytic model by Muench was chosen to the analysis. In the surveyed area, Kim-hae Goon where the egg positive rates were 56.2% and 61.2% (by Kim, 1974), the constant values of 'a' were found to be 0.051 and 0.089 respectively. In other words, the force of infection was 51, 89 per 1,000 susceptibles. The values of 'b' were found to be 0.006 and 0.005. This means that the rates of disappearance from egg positive cases to negative were 6 and 5 per annum per l,000 positive cases in above area. Therefore, the two catalytic curves were expressed by the following equations, y = 1.133 {e(-0.006t) - e(-0.051t)} and y = 1.047 {e(-0.005t) - e(-0.089t)} respectively. In the cases of North Kyong-sang Do and Ko-yang Goon, Kyong-gi Do where the egg positive rates of clonorchis shown as 27.7% and 15.2% by Shin (1964) and Kim (l974), the curves were expressed by y = 1.769 {e(-0.010t) - e( 0.034t)} and y = 2.857 {e(-0.020t) - e(-0.027t)} respectively. From the above mathematical analyses by age prevalence in clonorchiasis, it was considered that the mode of transmission of clonorchiasis in the surveyed area, Kim-hae Goon presented more rapid pattern than those of North Kyong-sang Do and Ko-yang Goon, Kyong-gi Do. PMID- 12902752 TI - [The Wormicidal Substances Of Fresh Water Fishes On Clonorchis Sinensis: I Preliminary Research On The Wormicidal Substance From Mucous Substance Of Carassius Carassius] AB - The present work was to observe the wormicidal effects of the external mucous substance of Carassius carassius and Pseudorasbora parva on cercaria, metacercaria (liberating) and adult of Clonorchis sinensis. The mucous substance which was extracted by ether was separated into various spots on the silica gel thin layer chromatography and separated into various fractions in the silica gel column chromatography, using petroleum ether(30%) and chloroform(70%) as solvent. 1. R(f) 0.952 value of the silica gel thin layer chromatography and the first fraction (reddish yellow) frorn the silica gel column chromatography of the ether extracts from C. carassius had the wormicidal effect on all of the developing stages of C. sinensis. Cercaria was killed within 14 minutes, metacercaria within 27 minutes and adult 2.5 hours by the first fraction. 2. But in the normal saline solution (control) and in the other fraction, cercaria and metacercaria were alive more than 24 hours and adult more than 72 hours. 3. The extracts from the mucous substance of P. parva that is well penetrated by cercaria had no wormicidal effect on them. PMID- 12902753 TI - [Study on the frequency distribution of the metacercarial density of Clonorchis sinensis in fish host, Pseudorasbora parva] AB - Since the pattern of the frequency distribution of a parasite within a host showed an overdispersed pattern, various statistical models such as Poisson, negative binomial and lognormal distributions have been applied on the population dynamics in host-parasite relations. The observed data on the number of metacercariae of Clonorchis sinensis in a suitable intermediate host, Pseudorasbora parva which were collected from an endemic area, Juk-Rim River, Kim Hae Goon, South Kyong-Sang Do were applied on the statistical models. The results obtained are as follows. 1. By the calculation of the raw data, 258(94.9%) out of 272 fish showed positive to the metacercarial infection of C. sinensis. The mean number of metacercaria in a fish was 335.1. The standard deviation was 250.6 and the mode was observed between 250 and 299. 2. The frequency distribution pattern of metacercariae of C. sinensis within the fish host in surveyed area was fitted to the lognormal distribution (0.052). It is found that Clonorchis sinensis is less prevalent in Seong-ju county than in the other counties of Kyungpook Province, Korea. PMID- 12902776 TI - [Studies On Pathological Changes Of The Liver In Abino Rats Infected With Clonorchis Sinensis] AB - The present study was undertaken to observe the pathologic changes of the liver in albino rats with Clonorchis sinensis. Twenty five rats given 50 metacercariae respectively by mouth were autopsied at 3 days (group 1), one week (group 2), 4 weeks (group 3), 8 weeks (group 4) and 12 weeks (group 5) of infection. The following results were obtained: 1. Parasites were seen in bile ducts of group 2, 3, 4, 5 and increased in number with duration of infection. 2. The pathologic changes in the liver were prominent in intrahepatic bile ducts. Epithelial cells of bile ducts showed definite atypia and proliferation resulting in pseudostratification in group. Stratification of metaplastic squamous cells and glandular proliferation were prominent in group 3. The epithelial cells were keratinized with syncitium and cribriform formation in group 4 and almost suggestive of adenomatous hyperplasia in group 5. 3. Periductal fibrosis seen in group 4 was considerable as well as mature with hyalinized connective tissue in group 5. 4. Heavy inflammatory cell infiltrations around the affected bile ducts in group 1 became smaller in number with duration of infection. The inflammatory cells consisted of the majority of eosinophils in group 1 and chiefly plasma cells, lymphocytes and mononuclear cells in group 5. 5. Veins in portal spaces dilated markedly in group 1 became less prominent with duration of infection. 6. Although portal fibrosis increased definitely in group 3 often extended into the hepatic lobules in group 4, the changes of hepatic cells, sinusoids and central veins were negligible. Above results suggest that clonorchiasis could be a factor in inducing primary carcinoma of liver in albino rats. PMID- 12902777 TI - [A Study On The Fine Structure Of Clonorchis Sinensis, A Liver Fluke: 1. The Body Wall And The Nervous System] AB - A morphological study on the ultrastructures of body wall and the nervous system of Clonorchis sinensis was conducted. For this study, liver flukes were collected from the liver of rabbit six months after the infection with metacercariae obtained from the fresh water fish, Pseudorasbora parva. The collected materials were washed with 0.85% saline solution and then immediately moved to cold 2.5% glutaraldehyde buffered with 0.1 M Millonig's phosphate buffer (pH 7.4). The materials were dissected into appropriate pieces in the fixative about 30 minutes after beginning of the fixation. Two hours later the materials were rinsed several times with the buffer and were fixed with cold, buffered 1% osmium tetroxide(OsO(4)) for 2 hours. The fully fixed tissue blocks were dehydrated in a series of graded concentrations of acetone and were embedded in Epon 812 mixture. Thin sections obtained from Sorvall MT-2 ultramicrotome were stained with uranyl acetate and Reynold's lead citrate. Observations of the sections were carried out with Hitachi HS-7S electron microscope. The following structures are newly identifed in a series of observations. 1. The mid-abdominal integument of the fluke is more thicker(~6 micrometer) than the dorsal side(about 3 micrometer). 2. Although it is so far known that only surface and epithelia of the suckers, pharynx, and the oesophagus are composed of integumental, anuclear layer, it is newly found that epithelium of the excretory pore also consists of integumental, anuclear layer. 3. There are lot of microvilli in the dorsal outer surface of the oral sucker. 4. A kind of ducts accumulated with unknown secretory granules are present within the parenchyrna around both of the oral suckers and the pharynx. 5. The nerve endings are distributed to the integument of the body where sensory hairs are projected outward. 6. The sensory hairs are rarely distributed throughout the whole integument, but a little more in the surroundings of both suckers. 7. The sensory hairs are attached to the base of the sheath by a circular septate desmosome. 8. No rootlet structure is observed but the upper part of the basal body is attached to the peripheral desmosome by transverse fibres. 9. The mitochondria and the vesicles are present within the hair cavity. 10. The nerve fibres are also found to be myelinated at least in main axons. PMID- 12902778 TI - [Epidemiological Study On Metagonimus Yokogawai Infection Along Boseong River In Jeonra Nam Do, Korea] AB - Prevalence study of Metagonimus yokogawai infection, survey of intermediate hosts of Metagonimus yokogawai and food habits of raw sweetfish among inhabitants along Boseong river (120 km), Jeonra Nam Do (province) were carried out. The results are summarized as follows: 1. Among 360 examined, the egg positive case were 128(35.6%); male 112(46.7%), female 16(13.3%). 2. The prevalence rates were different by geographical areas; 42.4% in down stream, 49.4% in middle stream and 13.3% in upper stream. 3. By ages, highest rate, 53.6~59.1% was observed among 30~49 group (male 69.6~73.0%, female 15.8~35.5%). 4. Eggs per gram of feces were 2,084 (200~12,280) in average; down stream 1,708, middle stream 2,590, upper stream l,550. 5. By social strata, the prevalence showed differences; 24.8% in primary schoolchildren, 78.4% in school teacher and local officers, 30.6% in general inhabitants. 6. Clonorchis sinensis infected cases in the areas were 21(5.8%) out of 360 examined, and 90.5% of them showed mixed infection with Metagonimus yokogawai. 7. Cercariae of Metagonimus yokogawai from the lst intermediate host, Semisulcospira sp., in down and middle streams were detected from 32 snails (5.9%) out of 545 examined. 8. All the 22 Plecoglossus altivelis which were caught from the down stream were infected with metacercariae of Metagonimus yokogawai. The number of the metacercariae detected was 22,650 per fish in average, and the number increased from August to October. 9. Experiences of raw sweetfish intake of the inhabitants were found in 208(58.4%) among 356 interviewed by questionaire form (male 76.7%, female 20.7%). 10. Means of acquisition of the sweetfish were mostly by purchase 68.8%, and seasonally in summer. 11. Reason of the raw sweetfish intake was due to the flavor, 83.2%. The fish were mixed in soy sause with vinegar before the intake. PMID- 12902779 TI - Effect Of Oxantel/pyrantel Pamoate Tablets Against Intestinal Nematodes In Korea. AB - A total of 48 subjects harbouring Trichuris trichiura, Ascaris lumbricoides, hookworm and/or Enterobius vermicularis were treated with oxantel/pyrantel pamoate tablets (100 mg of each) in a single dose of 20 mg base per kg of body weight. In all 37 cases of A. lumbricoides and 8 cases of A. duodenale infections cured completely, and the mean recovered worm-load was 3.8 with a range of 1-21 and 5.8 with a range of 3-9 respectively. In T. trichiura infection, 32(71.1%) of 45 cases cured completely and the average egg reduction rate was 90.8 per cent. The mean recovered worm-load was 14.6 with a rnage of 1-77 worms. On the other hand, in E. vermicularis infection, 37 (84.1%) of 44 patients were negative eggs by anal swab tests on the 10 to 11th days and 27 (61.4%) on the 20 to 21st days after treatment. The mean recovered worm-load was 17.7 with a range of 1-62. Only one patient with Trichostrongylus orientalis treated with this drug showed no eggs in his stools throughout the post-treatment examinations. Side effects were not noted in all treated cases. These findings demonstrate that a single dose of oxantel/pyrantel pamoate tablets can be considered a drug of choice for multiple infections with these nematodes. PMID- 12902780 TI - Reduced Single Dose Of Mebendazole In Treatment Of Ascaris Lumbricoides Infection. AB - The efficacy of reduced single doses of mebendazole against Ascaris lumbricoides infection was evaluated by cellophane thick smear and egg counting techinques, in two rural populations from December 1977 to March 1978. Total 191 infected cases were divided into five dose groups; 100 mg single dose, 200 mg single dose, 300 mg single dose, 400 mg and 600 mg in repeated doses. The cure rate of mebendazole in the treatment of Ascaris infection was remarkably high and satisfactory in every dose groups. The egg reduction rates were all over 98 percent in every dose groups. Side effects were observed during the treatment, mainly mild and transient gastrointestinal troubles, and not exacerbated by the increase of doses upto 300 mg in single dose. Above results confirmed that in case of mass chemotherapy of Ascaris infection, 100 mg or 200 mg of single dose treatment is sufficient to achieve the economic and acceptable method of treatment. PMID- 12902781 TI - [Study On The Chromosomal Proteins Of Fasciola Hepatica] AB - In attempt to investigate histone fractions and non-histones of parasites, nuclei were isolated from Fasciola hepatica by the procedure of Pogo et al. (1966). Histone fractions H1, H2a, H2b, H3 and H4 were prepared from isolated nuclei by the procedure of Johns (1964 and l967). The five histone fractions found in most tissues were also present in the Fasciola hepatica histones. These histone fractions were characterized by amino acid analysis and by polyacrylamide disc gel electrophoresis. Non-histone proteins were extracted from isolated Fasciola hepatica nuclei and separated by SDS-polyacrylamide gel electrophoresis. The results of the experiment were summarized as follows: 1. The yield of whole histone recovered was 2.47 mg per 1 g of Fasciola hepatica. 2. The yield of DNA was 1.02 mg per gm of tissues. Consequently the DNA to histone ratio was 1:2.44. 3. The relative amounts of five fractions, i.e., Hl, H2a, H2b, H3 and H4 were 19.96%, 26.48%, 29.60%, 12.56% and 14.37%, respectively. 4. Amino acid analysis of the individual histone fractions showed that the over-all compositions were similar but not identical to those of corresponding fraction from calf thymus. 5. It was found that histone H2b fraction of Fasciola hepatica contained detectable amounts of epsilon-N-monomethyllysine. No evidence for the presence of methylated lysine or other side-chain derivatives was reported on this histone fraction. 6. In SDS-polyacrylamide disc gel, it showed that 17 protein bands of nuclear acidic protein can be identified visually. PMID- 12902782 TI - [Immunological Tests By Anti-Free-Living Amoebas Serum Produced In Experimental Animals: I. Immobilization Of Free-Living Amoebas In Vitro By Rabbit Antiserum] AB - Rabbits were immunized with free-living amoebas by intravenous injections. The amoebas were Acanthamoeba culbertsoni and Naegleria fowleri and obtained by axenic cultivation in CGVS medium. Each rabbit received 10(6) of Acanthamoeba culbertsoni and 10(5) of Naegleria fowleri trophozoites respectively every other day in three doses and finally one booster dose at 1 week later. Antiserum was collected from thc following day of the booster injection up to 2 months period, and stored at -30 degrees C until use. The immobilization test was performed. One drop of amoeba suspension was mixed with the test serum on slide and observed the mobile state under microscope. 1. Maximal immobilizing phenomenon observed in 30 minutes and, then gradually recovered to normal state. 2. Inactivation of antiserum at 56 degrees C for 30 minutes did not affect the immobilization phenomenon. 3. The immobilization rates decreased by the serial dilution of antiserum. At dilution more than 1:8, the immobilization was almost the same as in the normal serum. 4. The immobilizing antibody in anti-Acanthamoeba culbertsoni rabbit serum showed highest titre in 3rd day after booster immunization and from first to 6th week in anti-Naegleria fowleri rabbit serum. 5. Cross matching of Acanthamoeba culbertsoni and Naegleria fowleri showed antigenic difference of the two species. It is suggested that the immobilization reaction may be of value as a supplementary test in the diagnosis of primary amoebic meningoencephalitis. PMID- 12902783 TI - An ecological study of Paragonimus in Malaysia. AB - An ecologic study on Paragonimus in Malaysia was attempted from May to September 1967. Seven streams located in various directions and distances from Kuala Lumpur were surveyed for the study of intermediate hosts, snail and crab. One Malayan village and one aborigine village where infected crabs were found, and two tuberculosis hospitals in K.L. were surveyed for the study of human population. Intradermal tests along with sputum or stool examination to detect human infection by Paragonimus were employed. Wild animals, only a few, were shot in the vicinity of the aborigine village and several domestic cats from the Malayan village were bought. These animals were autopsied and examined for adult Paragonimus. Among five species of crab collected from the study areas, only two species, Potamon jahorenes and Parathelphusa maculata were found to be infected with Paragonimus. P. maculata seemed to be better crab host for the Paragonimus because this species had higher infection rate and metacercarial density than the other in the very same area. Three out of seven streams had infected crabs and the infection rate as well as the infection intensity varied from one stream to another. Only avilable snail in the streams was identified as Brotia costula. The infection rate of the snail was very low, six snails out of 11,898, which is about the same rate reported from other countries. Infected snail, however, had thousands of rediae uncountable containing about twelve microcercocercariae in each redia, sufficient enough to maintain the life cycle of the parasite even with only a few infected snail, the amplifier. This is the first confirmed report on the snail host of Paragonimus from Malaysia where the existence of Paragonimus had been reported in 1923. The first trial to study human population by means of intradermal test, sputum and/or stool examinations in Malaysia showed no evidence of human infection of Paragonimus. The number of animals, wild and domestic, examined for natural infection was too small to draw any statement. These examined animals were all negative for adult Paragonimus. Even though more extensive studies on wild animals and human population may be necessary for the definite conclusion, the facts that infected crabs from jungle stream where human contacts are extreamely rare, and also highly infected crabs from the area where none of humans or domestic animals were infected, strongly suggest the life cycle of Paragonimus in this area may be maintained by wild animal hosts rather than by human host. The morphology of all stages of the parasite, the pattern of penetrating glands, flame cells and excretroy bladder of cercaria, lancet shaped single cuticular spines and 6 branched ovary of adult worm obtained from experimentally infected cat, and the shape of egg including all measurements agree well with the characteristics of Paragonimus westermani. PMID- 12902784 TI - A comparison of open reduction and internal fixation and primary total elbow arthroplasty in the treatment of intraarticular distal humerus fractures in women older than age 65. AB - OBJECTIVE: To compare open reduction and internal fixation (ORIF) with total elbow arthroplasty (TEA) for intraarticular distal humerus fractures in women older than 65 years of age. DESIGN: Retrospective review. SETTING: Information was obtained from a Level 1 trauma center with fellowship-trained traumatologists and a tertiary care center with fellowship-trained shoulder and elbow surgeons. PATIENTS: Patients were 24 women older than age 65 who sustained distal humerus fractures that required surgical treatment with clinical follow-up at a minimum of 2 years. All fractures were OTA classification 13.C2 or 13.C3. No patients were lost to follow-up. INTERVENTION: ORIF or TEA was the treatment method. MAIN OUTCOME MEASUREMENTS: The Mayo Elbow Performance score and the need for revision surgery were established as the means of patient evaluation. RESULTS AND CONCLUSIONS: Using the Mayo Elbow Performance score, the outcomes of the 12 patients treated with ORIF were as follows: 4 excellent, 4 good, 1 fair, and 3 poor (cases that required conversion to TEA). Outcomes of the 12 patients treated with TEA were as follows: 11 excellent and 1 good. There were no fair or poor outcomes in the TEA group. No patients treated with TEA required revision surgery. We believe TEA to be a viable treatment option for distal intraarticular humerus fractures in women older than age 65. This is particularly true for women with associated comorbidities, such as rheumatoid arthritis, osteoporosis, and conditions requiring the use of systemic steroids. PMID- 12902785 TI - Biomechanical comparison of posterior pelvic ring fixation. AB - OBJECTIVE: To determine relative stiffness of various methods of posterior pelvic ring internal fixation. DESIGN: Simulated single leg stance loading of OTA 61 Cl.2, a2 fracture model (unilateral sacroiliac joint disruption and pubic symphysis diastasis). SETTING: Orthopaedic biomechanic laboratory. OUTCOME VARIABLES: Pubic symphysis gapping, sacroiliac joint gapping, hemipelvis coronal plane rotation. METHODS: Nine different posterior pelvic ring fixation methods were tested on each of six hard plastic pelvic models. Pubic symphysis was plated. The pelvic ring was loaded to 1000N. RESULTS: All data were normalized to values obtained with posterior fixation with a single iliosacral screw. The types of fixation could be grouped into three categories based on relative stiffness of fixation: For sacroiliac joint gapping, group 1-fixation stiffness 0.8 and above (least stiff) includes a single iliosacral screw (conditions A and J), an isolated tension band plate (condition F), and two sacral bars (condition H); group 2-fixation stiffness 0.6 to 0.8 (intermediate stiffness) includes a tension band plate and an iliosacral screw (condition E), one or two sacral bars in combination with an iliosacral screw (conditions G and I); group 3-fixation stiffness 0.6 and below (greatest stiffness) includes two anterior sacroiliac plates (condition D), two iliosacral screws (condition B), and two anterior sacroiliac plates and an iliosacral screw (condition C). For sacroiliac joint rotation, group 1-fixation stiffness 0.8 and above includes a single iliosacral screw (conditions A and J), two anterior sacroiliac plates (condition D), a tension band plate in isolation or in combination with an iliosacral screw (conditions E and F), and two sacral bars (condition H); group 2-fixation stiffness 0.6 to 0.8 (intermediate level of instability) includes either one or two sacral bars in combination with an iliosacral screw (conditions G and I); group 3-fixation stiffness 0.6 and below (stiffest fixation) consists of two iliosacral screws (condition B) and two anterior sacroiliac plates and an iliosacral screw (condition C). DISCUSSION: Under conditions of maximal instability with similar material properties between specimens, differences in stiffness of posterior pelvic ring fixation can be demonstrated. The choice of which method to use is multifactorial. PMID- 12902786 TI - Retrograde fixation of distal femoral fractures: results using a new nail system. AB - OBJECTIVES: To investigate the management and outcome of distal femur fractures treated with retrograde nailing. DESIGN: Prospective. SETTING: Unfallkrankenhaus Berlin, level I trauma center. PATIENTS: A consecutive series of 47 patients with 48 fractures of the distal femur (37 fractures AO type A and 11 fractures type C) operated on between May 1999 and June 2000. OUTCOME MEASURES: Outcome was assessed by using standard radiographic criteria of time to union, incidence of infection, malunion, and knee function (Leung score). RESULTS: After an average time of 33 months (range 12-37 months), 44 patients were reexamined. Three patients were lost to follow-up. The average age was 44 years (range 17-92 years). Of patients, 19 sustained polytrauma, and 10 had associated soft tissue damage. A total of 34 patients underwent primary definitive osteosynthesis within 12 hours after trauma. All fractures healed after an average of 12.6 weeks (range 9-17 weeks). Seven complications were noted-three related to severity of injury (one deep venous thrombosis, two leg length shortenings of 1.5 cm and 2.5 cm) and four related to the operation (insufficient counterboring of the nail in two patients, one malreduction, one iatrogenic fracture of femur shaft). There was no relevant difference between type A and type C fractures in functional, clinical, or radiographic outcomes. CONCLUSIONS: Retrograde nailing is recommended as an alternative method to plate osteosynthesis in stabilizing distal femoral fractures, particularly in type C fractures. PMID- 12902788 TI - Pin tract infection with contemporary external fixation: how much of a problem? AB - OBJECTIVE: To determine the incidence of pin tract infection.DESIGN Retrospective chart review. SETTING: Level 1 trauma center in an urban community. PATIENTS: A total of 285 patients with 285 fractures over a 4-year period (1997-2001). INTERVENTION: External fixation. MAIN OUTCOME MEASUREMENT: Incidence of pin tract infection. RESULTS: Of 285 fractures, 32 (11.2%) were complicated by infection. The incidence of infection according to montage was 3.9% (3/77) for ring fixators, which was significantly different (P < 0.04) from the 12.9% incidence (23/178) for unilateral fixators and the 20.0% incidence (6/30) for hybrid fixators (P = 0.004). The incidences of pin tract infection for the unilateral fixator group and the hybrid fixator group were not significantly different. CONCLUSIONS: Patients with hybrid external fixators had a similar risk of pin tract infection as patients who had unilateral fixators. The infection rate in the ring fixator group was significantly lower than the hybrid external and unilateral fixator groups. PMID- 12902787 TI - Proximal tibial fracture stability with intramedullary nail fixation using oblique interlocking screws. AB - OBJECTIVES: The purpose of this study was to evaluate the mechanical stability of oblique interlocking screws in supplementing intramedullary nail fixation of high proximal tibial fractures. DESIGN: In vitro experimental testing. SETTING Orthopaedic biomechanics laboratory, Sunnybrook and Women's College Health Sciences Center. PARTICIPANTS: Ten paired fresh-frozen human cadaver tibiae. INTERVENTION: One tibia of each pair was randomized to be instrumented with an intramedullary nail (M/DN; Zimmer, Warsaw, Indiana), while the other was stabilized with a 13-hole stainless steel lateral tibial head plate (Synthes AO/ASIF). Specimens were tested in varus-valgus (v/v), flexion-extension (f/e) and torsion, before and after a 2-cm gap osteotomy was performed in the proximal segment. Testing of the nailed tibiae was performed with and without oblique proximal screws. Bone density was physically determined by removing a core of trabecular bone from the distal end of each tibia following testing. MAIN OUTCOME MEASUREMENT: Biomechanical construct stability. RESULTS: The addition of the proximally placed oblique screws increased the stability of the nail construct in v/v by 50% (6.8 mm, P < 0.05), in f/e by 47% (7.2 mm, P < 0.05), and in torsion by 18% (3.0 degrees, P < 0.05). There was no significant difference observed between the stability of the intramedullary nail construct with oblique screws and the plated construct. Trabecular bone density had a significant effect in reducing stability (P < 0.05) in nail and plate fixation. CONCLUSION: The addition of oblique interlocking screws significantly improves the stability of a nailed proximal tibia fracture and provides comparable stability to a plate osteosynthesis. PMID- 12902789 TI - The new injury severity score is a better predictor of extended hospitalization and intensive care unit admission than the injury severity score in patients with multiple orthopaedic injuries. AB - OBJECTIVE: To compare the scoring efficacy of the injury severity score (ISS) and the new injury severity score (NISS) in predicting extended hospital length of stay (LOS) and intensive care unit (ICU) admission and to determine the effect of multiple orthopaedic injuries (MOI) on the discrepancies between the ISS and NISS and their impact on extended LOS and ICU admission. DESIGN: Prospective cohort study. SETTING: Level 1 university trauma center. PATIENTS: All consecutive trauma admissions during the 2-year period ending on December 31, 2000, with age older than 14 years and survival greater than 24 hours were entered into the study. MAIN OUTCOMES: Extended (>or=10 days) hospital LOS and ICU admission. RESULTS: Of 3,100 patients, 7.5% had a higher NISS than ISS, spent more days in the hospital (22 versus 8; P < 0.001) and in the ICU (3.4 versus 0.1; P < 0.001), and had a higher mortality rate (8% versus 1.2%; P < 0.001) than patients with identical NISS and ISS. The NISS was found to be more predictive of longer (>or=10 days) LOS (receiver operating characteristic [ROC] NISS = 0.794, ISS = 0.782; P < 0.0001) and ICU admission (ROC NISS = 0.944, ISS = 0.918; P < 0.0001). The multivariate predictive model including NISS showed a better goodness of fit compared with the same model that included ISS. Patients with discrepant scores (NISS > ISS) spent a longer time in the hospital and in the ICU in addition to having an increased frequency of ICU admission. In 61% of the cases, MOI were responsible for the discrepant (NISS > ISS) scores. CONCLUSIONS: MOI have a significant effect on trauma outcomes such as LOS and ICU admission. The recognition of this high-risk group is not possible using the traditional ISS alone from retrospective or prospective databases. Considering its easier calculation and better predictive power, it is suggested that the NISS should replace the traditional ISS in trauma outcome research. PMID- 12902791 TI - Retrograde removal of an incarcerated solid titanium femoral nail after subtrochanteric fracture. AB - Intramedullary nailing with a solid titanium nail can result in significant bone ingrowth. Failure of an implant requires removal for replacement with another device. Previous methods have included use of extractors for cannulated nails. When extractors are not available or are inadequate, a simple retrograde push-out method can be used as described here. PMID- 12902792 TI - Pelvic C-clamp in a pediatric patient. AB - The pelvic C-clamp was designed to provide posterior stability temporarily to pelvic ring injuries associated with massive venous hemorrhage in adult patients. Pediatric pelvic ring injuries are uncommon, but when unstable injuries occur they may be coupled with visceral injuries and lethal hemorrhage. 1, 2 Anterior external frame fixators have been described in the treatment of these injuries in pediatric patients with few adverse outcomes and good long-term results. Although they provide adequate fixation, anterior frame application is complicated, time consuming, and cumbersome as an emergency application. The pelvic C-clamp eliminated these problems, while providing fixation points that do not violate areas of cartilage or physeal growth. We present a case demonstrating that the pelvic C-clamp is a safe form of temporary fixation in a skeletally immature patient with a posterior pelvic ring injury associated with massive hemorrhage. PMID- 12902790 TI - Temporary bridge plating of the medial column in severe midfoot injuries. AB - Severe crush injuries to the midfoot often involve comminuted cuneiform or tarsal navicular fractures. Treatment principles for the bony injury of the crushed midfoot include maintenance of the medial column length and alignment, as well as appropriate stable fixation after open or closed fracture reductions. This is especially important because outcomes after midfoot injuries are related to the stability of the medial longitudinal arch of the foot. Treatment options include closed reduction and isolated K-wire fixation, limited open reduction and internal fixation with K-wires, screw fixation directed from the navicular to the cuneiforms, spanning external fixation between the talus and the first metatarsal, or combinations of these techniques. Limited internal fixation combined with external fixation may be difficult or impossible in comminuted fractures secondary to the small size and large number of bony fragments. Also, the external fixator is a potential source of pin tract infections. We propose a temporary internal bridge plating technique of the medial column of the foot using an 8- to 10-hole, 2.7-mm reconstruction plate between the talar neck and the first metatarsal, which may provide adequate temporary internal stabilization until bony healing occurs. PMID- 12902793 TI - A combined proximal and distal dislocation of two adjacent metatarsals: double floating metatarsal bones (second-third). AB - A rare foot injury consisting of an irreducible, closed combined distal and proximal articulation dislocation of two adjacent metatarsals (second and third), called a "double floating metatarsal," was encountered in a 29-year-old man. The anatomy, mechanism of injury, and sequence of the surgical procedure are presented and discussed. PMID- 12902794 TI - Posterior dislocation of the elbow with concomitant fracture of the proximal ulnar diaphysis and radial head: a complex variant of the posterior monteggia lesion. PMID- 12902795 TI - Achilles tendon rupture associated with injury of the calcaneofibular ligament. AB - A 49-year-old man collided against an infielder when he slid into second base during a recreational baseball game. He was unable to continue in the game due to diffuse pain and swelling of his hindfoot. A rupture of the Achilles tendon was diagnosed incidentally on palpation and observation of a positive Thompson's squeeze test. Subcutaneous hemorrhage at the lateral aspect of the heel and a small bone fragment under the lateral malleolus on an anteroposterior plain radiograph indicated a fracture of the calcaneal wall. At surgery, a complete rupture of the Achilles tendon and an avulsion of the calcaneofibular ligament from the calcaneal wall were seen. Both injuries were surgically repaired, and the patient subsequently did well. The mechanism of injury was thought to be impact hyperdorsiflexion of the ankle with rupture of the Achilles tendon accompanied by an inversion injury. Using a literature search, it was found that this combination of injuries has not been previously reported. PMID- 12902796 TI - Letters to the editor. PMID- 12902797 TI - The effects of protease inhibitors and nonnucleoside reverse transcriptase inhibitors on p-glycoprotein expression in peripheral blood mononuclear cells in vitro. AB - Several antiretroviral compounds have been shown to be substrates for the efflux protein P-glycoprotein (P-gp) although few studies have investigated the effects of drug on expression of this protein. Here, an in vitro system has been adopted to investigate the effects of protease inhibitors (PIs) and nonnucleoside reverse transcriptase inhibitors (NNRTIs) on P-gp expression in peripheral blood mononuclear cells (PBMCs). PBMCs isolated from healthy volunteers were incubated with 10 or 100 microM PI (saquinavir, ritonavir, lopinavir, indinavir, nelfinavir, amprenavir) or 10 microM NNRTI (efavirenz, nevirapine) for 72 hours. Surface P-gp expression was measured by flow cytometry and compared with vehicle incubated controls. Toxicity was assessed by MTT assay and the effects of each compound were compared between individuals with differing genotypes at position 3435 of exon 26 of MDR1, which was assigned by restriction fragment length polymorphism. Significant increases in median P-gp expression were observed following incubation with 10 microM nelfinavir (10.2 versus 6.7% P-gp-positive cells) and efavirenz (10.0 versus 6.7% P-gp-positive cells). No significant differences in induction were observed between genotypes (CC, CT, TT). Following incubation with 100 microM PI, significant upregulation of P-gp occurred except with amprenavir. However, nelfinavir, ritonavir, and lopinavir caused marked toxicity, indicating that at higher concentrations, the increase in P-gp may be at least partially related to a stress response. These results indicate the potential of some PIs and NNRTIs to induce P-gp expression in PBMCs in vitro. PMID- 12902798 TI - Viral dynamics and their relations to baseline factors and longer term virologic responses in treatment-naive HIV-1-infected patients receiving abacavir in combination with HIV-1 protease inhibitors. AB - From a study of 71 HIV-1-infected patients receiving abacavir in combination with 1 of 5 different HIV-1 protease inhibitors (indinavir, ritonavir, saquinavir, nelfinavir, or amprenavir), we found that the baseline HIV-1 RNA levels were highly predictive of the viral decay rates. The baseline HIV-1 RNA levels were negatively correlated with the first phase viral decay rates (r = -0.77, P < 0.001) and positively correlated with the second phase viral decay rates (r = 0.68, P < 0.001). In addition, the first phase viral decay rate was positively correlated with CD4+ cell increases. No significant correlation was found between viral decay rates and longer term (24 weeks) virologic responses, and no difference in viral decay rates was found among the 5 study regimens. These data suggest that the potency of the 5 treatment regimens was similar and was not predictive of long-term virologic failure. PMID- 12902800 TI - An objective lipodystrophy severity grading scale derived from the lipodystrophy case definition score. AB - HIV lipodystrophy can be objectively diagnosed using a score derived from the 10 parameters in the HIV lipodystrophy case definition (LDCD). Lipodystrophy severity remains subjectively determined by physical examination and patient assessment. Regional dual-energy x-ray absorptiometry (DEXA) and single-slice abdominal computed tomography (CT) scanning are objective but are gender dependent body composition measures. The LDCD score may provide a means of generating an objective and lipodystrophy grading/severity scale applicable to both men and women. Total and regional clinical lipodystrophy severity scores (generated using the HIV Outpatient Study [HOPS] scale: nil (0), mild (1), moderate (2), and severe (3) lipoatrophy or fat accumulation in 8 body regions) were correlated with objective measures of LD (LDCD score, DEXA, abdominal CT) and metabolic (lipid, glycemic, acid-base) parameters known to correlate significantly with lipodystrophy severity. Analysis was based on 417 lipodystrophic adults and 371 controls recruited to the HIV LDCD study. Correlation coefficients were used to compare physician and patient assessments (rPhysician, rPatient) with objective LD measures and metabolic parameters. The strongest objective correlate of total clinical lipodystrophy severity was the LDCD score (rPhysician = 0.641 [95% CI, 0.584-0.698]; rPatient = 0.620 [95% CI, 0.561-0.678]), whereas the strongest imaging correlate (trunk:limb fat ratio on DEXA) was significantly lower (rPhysician = 0.483 [95% CI, 0.420-0.546]; rPatient = 0.475 [95% CI, 0.412-0.538]; P < 0.001). The LDCD score also yielded significantly greater correlations with 7 of the 8 metabolic parameters than did clinical lipodystrophy severity scores. Based on quartiles of physician-rated severity, the LDCD scores were categorized to allow for rating of lipodystrophy as absent (LDCD score < 0), grade 1 (0-9.9), grade 2 (10-14.9), grade 3 (15 22.9), and grade 4 (>or=23). In conclusion, the LDCD score is the best objective measure of lipodystrophy severity and, in contrast to DEXA and CT, it is also gender independent. Subjective assessment of lipodystrophy severity could possibly be abandoned in cross-sectional studies. The LDCD score and its derived lipodystrophy grading scale merit prospective evaluation. PMID- 12902799 TI - Leptin replacement therapy but not dietary polyunsaturated fatty acid alleviates HIV protease inhibitor-induced dyslipidemia and lipodystrophy in mice. AB - A major complication associated with the use of protease inhibitors (PIs) in treatment of HIV-infected patients is lipid abnormalities including dyslipidemia, lipodystrophy, and liver steatosis. Previous studies revealed that these abnormalities are associated with PI-induced accumulation of activated sterol regulatory element binding proteins (SREBPs) in the nucleus of liver and adipose tissues, resulting in constitutive activation of lipid metabolism genes. This study used the mouse model to determine the potential of polyunsaturated fatty acid (PUFA) diet or leptin replacement therapy to alleviate these PI-induced metabolic abnormalities. Results showed that feeding C57BL/6 mice with a PUFA rich diet failed to normalize plasma cholesterol and triglyceride levels in ritonavir-treated mice. The PUFA-rich diet also had no effect on ritonavir induced interscapular fat accumulation and liver steatosis. In contrast, daily administration of leptin significantly reversed the elevated plasma cholesterol level induced by ritonavir. Leptin replacement therapy also significantly reduced the ritonavir-induced interscapular fat mass and improved liver steatosis. Taken together, these data suggest that PI-induced lipid abnormalities, especially dyslipidemia, lipodystrophy, and liver steatosis, may be reduced with leptin replacement therapy. PMID- 12902801 TI - The effect of HAART and HCV infection on the development of hyperglycemia among HIV-infected persons. AB - OBJECTIVE: To examine the prevalence and incidence of hyperglycemia among HIV infected patients by hepatitis C virus (HCV) infection and type of highly active antiretroviral therapy (HAART). DESIGN Retrospective cohort analysis of 1230 persons on their first HAART regimen who had at least 1 random glucose measurement before and during antiretroviral therapy. METHODS: The prevalence of hyperglycemia and the incidence of hyperglycemia were compared among persons with and without HCV infection while on a protease inhibitor (PI)-containing HAART regimen, a nonnucleoside reverse transcriptase inhibitor (NNRTI)-containing regimen, or a regimen that contained both a PI and an NNRTI. Hyperglycemia was defined as either 2 random glucose levels > 11.1 mM (200 mg/dL) or documentation of the diagnosis of diabetes in the medical record. RESULTS: The prevalence of hyperglycemia was significantly higher in HCV-coinfected (5.9%) than HCV uninfected persons (3.3%, P = 0.02). Among persons receiving HAART, both HCV coinfection (adjusted relative hazard [ARH], 2.28; 95% CI, 1.23-4.22) and PI use (ARH, 5.02; 95% CI, 1.39-18.16) were independent risk factors of developing hyperglycemia. The incidence of hyperglycemia was highest among HCV-coinfected persons receiving a PI (5.6 cases per 100-person years) and only 1 person who was neither HCV-infected nor receiving a PI developed hyperglycemia. CONCLUSION: In this urban HIV cohort, the risk of hyperglycemia was increased in HCV-coinfected patients and those prescribed a PI. PMID- 12902802 TI - Progression of HIV disease among women following delivery. AB - OBJECTIVE: To assess patterns of HIV disease progression among HIV-1-infected women following delivery. METHODS: Four hundred ninety-seven women enrolled in PACTG 185, a phase 3 trial of passive immunoprophylaxis in addition to zidovudine (ZDV) for the prevention of perinatal transmission, were included. Visits occurred twice during pregnancy; at delivery; and at 12, 26, 48, and 78 weeks postpartum. Repeated-measures linear regression and proportional hazards models were applied. RESULTS: Trial treatment (HIV hyperimmune globulin vs. immune globulin) was not related to postpartum disease progression. Longitudinal analysis of HIV-1 RNA demonstrated stable levels during pregnancy, significantly increased HIV-1 RNA by 12 weeks postpartum even on stable therapy, and a gradual increase thereafter. Changes in CD4+ lymphocyte percentage over 18 months of follow-up were similar for women continuing or stopping ZDV postpartum. Compared with those receiving no therapy, the hazard ratio for AIDS or death among women who received monotherapy postpartum was 0.52 (95% confidence interval [CI]: 0.25 1.04), 0.17 (CI: 0.06-0.49) for women who received combination therapy, and 0.24 (CI: 0.06-1.01) for women who received highly active antiretroviral therapy. CONCLUSIONS: RNA levels increased significantly from delivery to 12 weeks postpartum. Changes in HIV-1 RNA and CD4+ lymphocyte percentage were similar among women continuing or stopping therapy after delivery, and response to antiretroviral therapy was as expected postpartum. PMID- 12902803 TI - Lopinavir plasma concentrations and changes in lipid levels during salvage therapy with lopinavir/ritonavir-containing regimens. AB - OBJECTIVE: To determine whether an association existed between lopinavir (LPV) plasma concentrations and changes in lipid levels. DESIGN: A prospective, nonrandomized study. SUBJECTS: HIV-infected subjects with virologic failure on protease inhibitor-containing regimens. Twenty-two consecutive patients were enrolled, 19 completed 24 weeks of treatment, and 16 completed the full 48-week study period. INTERVENTION Patients were treated with LPV/ritonavir (LPV/r) in combination with other antiretroviral agents. Subjects were evaluated at baseline and weeks 4, 8, 12, 24, 36, and 48. LPV trough plasma concentrations and lipid levels were measured. RESULTS: LPV trough concentrations were higher in patients experiencing grade 3 or higher lipid elevations (mean [SD]: 9.71 microg/mL (5.62) vs. 6.09 microg/mL (3.83); P = 0.002) and in those developing grade 2 or higher hypercholesterolemia (mean [SD]: 8.48 microg/mL [4.64] vs. 5.71 microg/mL [3.94]; P = 0.003). All patients developing grade 2 or higher cholesterol elevation had an LPV trough concentration at week 4 greater than 8 microg/mL. Significant positive correlations were found between LPV trough concentrations and changes in triglyceride and cholesterol levels. CONCLUSIONS: In patients receiving salvage therapy with LPV/r, there is an association between LPV plasma concentrations and lipid changes. Patients achieving higher LPV trough concentrations may be at greater risk of experiencing dyslipidemia. Further investigations are warranted to support a direct cause and effect relationship. PMID- 12902804 TI - Interleukin-7 and immunologic failure despite treatment with highly active antiretroviral therapy in children perinatally infected with HIV-1. AB - Baseline interleukin-7 (IL-7) level has been proposed as a predictor of immunologic response to antiviral therapy, and the use of exogenous IL-7 has been suggested in the treatment of HIV-infected patients. Therefore, we investigated the relationships between IL-7 serum levels and immunologic outcome of highly active antiretroviral therapy (HAART) in 24 children perinatally infected with HIV-1. IL-7 serum levels were evaluated by enzyme-linked immunosorbent assay before switching antiretroviral treatment from double therapy to HAART and then again after 12 weeks of HAART. Differences were analyzed between children with immunologic failure and those without. At baseline, IL-7 levels were significantly higher in HIV-1-infected children than in 24 healthy age-matched uninfected children (16.6 +/- 8.7 vs. 9.5 +/- 2.4 pg/mL, respectively; P < 0.001). IL-7 levels inversely correlated with CD4 T-lymphocyte percentage both at baseline (r = -0.71, P < 0.001) and after 12 weeks of HAART (r = -0.49, P = 0.01). We observed no correlation between IL-7 levels and viral load (r = 0.29, P = 0.17 at baseline; r = 0.30, P = 0.15 after 12 weeks). Baseline IL-7 levels were similar in HIV-1-infected children with or without subsequent immunologic failure (15.1 +/- 8.8 vs. 17.5 +/- 8.7 pg/mL, respectively; P = 0.75). After 12 weeks of HAART, IL-7 levels were 21.6 +/- 13.5 pg/mL in children with immunologic failure (P = 0.08 when compared with baseline levels) and 8.2 +/- 3.8 pg/mL in children without immunologic failure (P = 0.002 when compared with baseline levels). IL-7 levels were significantly higher (P = 0.003) in children with immunologic failure than in those without. Findings indicate that baseline IL-7 serum levels do not predict immunologic outcome of HAART and that immunologic failure occurs even in the presence of high IL-7 serum levels, thus suggesting no benefit from therapy with exogenous IL-7. PMID- 12902805 TI - Effect of antioxidants on glucose metabolism and plasma lipids in HIV-infected subjects with lipoatrophy. AB - Ten HIV-infected nucleoside reverse transcriptase inhibitor-treated subjects with lipoatrophy or sustained hyperlactatemia were given antioxidants: vitamins C, E, and N-acetyl cysteine. After 24 weeks, anthropometrics did not change significantly, except for a modest decrease in the waist-to-hip ratio. Fasting low-density lipoprotein cholesterol trended toward lower values. Fasting glucose significantly increased along with a significant increase in homeostatic model assessment values, reflecting an increase in insulin resistance. Controlled trials are required to evaluate directly the effects of these agents on lipid and glucose metabolism. PMID- 12902806 TI - Contraception use, family planning, and unprotected sex: few differences among HIV-infected and uninfected postpartum women in four US states. AB - To describe pregnancy intentions and contraceptive use among a postpartum sample of women with and at risk for HIV infection, 258 HIV-seropositive and 228 HIV seronegative women were recruited from prenatal clinics in 4 US states between June 1996-November 1998. Participants completed interviews at 24-40 weeks' gestation and at 6 months postpartum. At the 6-month interview, 78% of women reported vaginal sex, and 2% were pregnant. Among those not pregnant, 86% said that there was no likelihood of a pregnancy in the next 6 months. Condom use was reported by 68% of sexually active women; 65% of users reported consistent use. Those with HIV were more likely to report condom use, more likely to report condom use consistency, and less likely to report use of oral contraceptives than women without HIV (P < 0.05). In multivariate analysis, inconsistent condom use was associated with postpartum alcohol use (odds ratio [OR] 2.80; 95% CI = 1.34 5.84), with the respondent stating that a pregnancy would not be emotionally upsetting (OR 3.06; 95% CI = 1.41-6.59) and reporting an intention to terminate a pregnancy if one were to occur (OR 3.47; 95% CI = 1.58-7.60). HIV-seropositive women who had at least 1 child with HIV infection were less likely than seronegative women to report inconsistent condom use (OR 0.15; 95% CI = 0.03 0.76). Few differences were detected in reproductive behaviors as a function of HIV serostatus, although both cohorts engaged in unprotected sex. Counseling to decrease sexual risk behaviors should begin prior to or early in the postpartum period and include discussion of both reproductive and disease transmission issues. PMID- 12902807 TI - Clinical and immunologic progression in HIV-infected US women before and after the introduction of highly active antiretroviral therapy. AB - OBJECTIVE: To examine factors associated with clinical and immunologic HIV disease progression in a cohort of US women. DESIGN: Analysis of data from a prospective, longitudinal, case-control study of HIV-infected women followed every 6 months for 7 years. SETTING: Four urban clinical centers in the United States. PARTICIPANTS: 648 HIV-infected women who did not have AIDS at time of entry into the study. MEASUREMENTS: Structured clinical and behavioral interviews; protocol-directed physical examinations; CD4 lymphocyte counts; plasma HIV RNA; infectious pathogen serologies. RESULTS With 2304 women-years of follow-up, 46.1% of the women developed AIDS; however, 93.3% of the diagnoses were based on CD4 counts dropping to <200 cells/mm(3). Only 10.6% of the women with CD4 counts <200 cells/mm(3) developed an opportunistic infection. Baseline CD4 count was the strongest predictor of subsequent clinical progression. Illicit substance use, multiple pregnancies, demographic variables, and other infections were not associated with progression. Among women with CD4 counts >500 cells/mm(3) at baseline, those who were anemic or had hepatitis C were more likely to progress to AIDS. By the end of the study, only 52% of the participants were on highly active antiretroviral therapy (HAART). CONCLUSIONS: Despite underutilization of HAART in this multicenter cohort of urban women, opportunistic infections were uncommon, despite CD4 declines. PMID- 12902808 TI - Performance characteristics of a new less sensitive HIV-1 enzyme immunoassay for use in estimating HIV seroincidence. AB - Less sensitive (LS) HIV-1 enzyme immunoassays (EIAs) have significantly improved the quantity and quality of HIV surveillance data. The first LS-HIV-1 EIA, the Abbott 3A11-LS, provided reliable incidence data, but the assay required specialized equipment, and the lack of available reagents made testing difficult. This study evaluated the use of an alternate assay, a modified version of the Vironostika HIV-1 EIA (Vironostika-LS), to be used for LS testing. The Vironostika-LS has similar performance characteristics to the Abbott 3A11-LS with additional advantages. This 96-well formatted assay is commonly found in public health laboratories for routine HIV-1 testing and can be used with both serum and dried blood spot specimens. The estimated mean time from seroconversion (defined using a standardized optical density cutoff of 1.0) with the Vironostika-LS was 170 days (95% CI, 145-200 days). When the Vironostika-LS was applied to a matched serum set previously tested with the Abbott 3A11-LS, the Vironostika-LS accurately identified 97% of specimens with recent or long-standing HIV infection. The paper also reports Vironostika-LS quality control guidelines and the results from 3 rounds of proficiency testing. PMID- 12902809 TI - Are there gender and race differences in cellular immunity patterns over age in infected and uninfected children born to HIV-infected women? AB - This study investigated whether age-related patterns of immunologic markers in 1488 uninfected (9789 measurements) and 186 infected (3414 measurements) children differed by gender and race. CD4+, CD8+, and absolute lymphocytes by HIV infection status, gender, and race were assessed using linear mixed-effects natural cubic spline models, allowing for prematurity and maternal CD4+ cell count. In uninfected children, levels of all 3 markers peaked twice in the first few months of life, declining to adult levels by around 8 years of age; uninfected boys and uninfected black children had significantly reduced CD4+ and absolute lymphocyte counts; the gender difference was especially pronounced in black children. Infected children had substantially lower levels and distinctly different patterns; with, e.g., by age 6 months CD4+ cell counts nearly 1200 per mm3 lower than in uninfected infants. Levels also significantly differed by gender and race for infected children, although for gender in the opposite direction. The gender and race differences in CD4+ levels were not explained by a general lymphocytosis nor were they confounded by treatment. These substantial differences in immunologic markers may reflect underlying genetic influence on the cellular immune system and may have implications for clinical decisions about therapeutic management. PMID- 12902810 TI - The value of screening for sexually transmitted diseases in an HIV clinic. AB - Because bacterial sexually transmitted diseases (STDs) facilitate HIV transmission, screening for and treatment of STDs among HIV-infected persons should prevent HIV spread to partners. Before screening programs for gonorrhea and Chlamydia infection should be widely established in HIV clinics, it is useful to know the prevalence of these infections. This study analyzed the results of a urine-based screening program for gonorrhea and Chlamydia in a New Orleans HIV clinic and compared the positivity rates to the prevalence in the local community. Among persons screened in the HIV clinic, 1.7% (46/2629) had gonorrhea and 2.1% (56/2629) had Chlamydia infection. Among persons aged 18-29 years, the test positivity for gonorrhea was similar in the HIV clinic to that of persons in sociodemographically similar community samples (3.1 versus 2.4%, adjusted odds ratio 1.6, P = 0.11) and the test positivity for Chlamydia infection was lower (5.4% versus 10.5%, adjusted odds ratio 0.6, P < 0.01). Based on a previously published mathematical model, it was estimated that treatment of all 46 gonorrhea and 56 Chlamydia infections in the HIV clinic may have averted 9 HIV infections among sex partners and saved far more in future medical costs than the cost of the screening. Routine screening for gonorrhea and Chlamydia infection should be considered in HIV clinics. PMID- 12902811 TI - High prevalence of HIV infection among women and their children in Henan Province, China. PMID- 12902812 TI - Lack of methadone dose alterations or withdrawal symptoms during therapy with lopinavir/ritonavir. PMID- 12902813 TI - Pilot study of directly observed therapy in highly nonadherent HIV-infected patients in an urban community-based institution. PMID- 12902814 TI - Changes in metabolic profile among antiretroviral-naive patients initiating protease inhibitor versus non-protease inhibitor containing HAART regimens. PMID- 12902815 TI - Increased human T-lymphotropic virus type II proviral load following highly active retroviral therapy in HIV-coinfected patients. PMID- 12902832 TI - Cerebral microvessel responses to focal ischemia. AB - Cerebral microvessels have a unique ultrastructure form, which allows for the close relationship of the endothelium and blood elements to the neurons they serve, via intervening astrocytes. To focal ischemia, the cerebral microvasculature rapidly displays multiple dynamic responses. Immediate events include breakdown of the primary endothelial cell permeability barrier, with transudation of plasma, expression of endothelial cell-leukocyte adhesion receptors, loss of endothelial cell and astrocyte integrin receptors, loss of their matrix ligands, expression of members of several matrix-degrading protease families, and the appearance of receptors associated with angiogenesis and neovascularization. These events occur pari passu with neuron injury. Alterations in the microvessel matrix after the onset of ischemia also suggest links to changes in nonvascular cell viability. Microvascular obstruction within the ischemic territory occurs after occlusion and reperfusion of the feeding arteries ("focal no-reflow" phenomenon). This can result from extrinsic compression and intravascular events, including leukocyte(-platelet) adhesion, platelet-fibrin interactions, and activation of coagulation. All of these events occur in microvessels heterogeneously distributed within the ischemic core. The panorama of acute microvessel responses to focal cerebral ischemia provide opportunities to understand interrelationships between neurons and their microvascular supply and changes that underlie a number of central nervous system neurodegenerative disorders. PMID- 12902833 TI - Antiactin-targeted immunoliposomes ameliorate tissue plasminogen activator induced hemorrhage after focal embolic stroke. AB - Thrombolytic stroke therapy with tissue plasminogen activator (tPA) is limited by serious risks of intracerebral hemorrhage. In this study, the authors show that a novel antiactin-targeted immunoliposome significantly reduced tPA-induced hemorrhage in an established rat model of embolic focal stroke. Spontaneously hypertensive rats were subjected to focal ischemia using homologous blood clot emboli. Delayed administration of tPA (10 mg/kg, 6 hours after ischemia) induced intracerebral hemorrhage at 24 hours. In control rats treated with tPA plus vehicle, hemorrhage volumes were 9.0 +/- 2.4 uL (n = 7). In rats treated with tPA plus antiactin immunoliposomes, hemorrhage volumes were significantly reduced to 4.8 +/- 2.7 uL (n = 8, P < 0.05). No significant effects were seen when rats were treated with tPA plus a nontargeted liposome (7.8 +/- 2.1 uL, n = 9). Fluorescent immunohistochemistry showed that rhodamine-labeled targeted liposomes colocalized with vascular structures in ischemic brain that stained positive for endothelial barrier antigen, a marker of cerebral endothelial cells. These data suggest that immunoliposomes may ameliorate vascular membrane damage and reduce hemorrhagic transformation after thrombolytic therapy in cerebral ischemia. PMID- 12902834 TI - Metabolic, neurochemical, and histologic responses to vibrissa motor cortex stimulation after traumatic brain injury. AB - During the prolonged metabolic depression after traumatic brain injury (TBI), neurons are less able to respond metabolically to peripheral stimulation. Because this decreased responsiveness has been attributed to circuit dysfunction, the present study examined the metabolic, neurochemical, and histologic responses to direct cortical stimulation after lateral fluid percussion injury (LFPI). This study addressed three specific hypotheses: that neurons, if activated after LFPI, will increase their utilization of glucose even during a period of posttraumatic metabolic depression; that this secondary activation results in an increase in the production of lactate and a depletion of extracellular glucose; and that because cells are known to be in a state of energy crisis after traumatic brain injury, additional energy demands resulting from activation can result in their death. The results indicate that stimulating to levels eliciting a vibrissa twitch resulted in an increase in the cerebral metabolic rate for glucose (CMR(glc); micromol.100 g(-1).min(-1)) of 34% to 61% in the sham-operated, 1-hour LFPI, and 7-day LFPI groups. However, in the 1-day LFPI group, stimulation induced a 161% increase in CMR(glc) and a 35% decrease in metabolic activation volume. Extracellular lactate concentrations during stimulation significantly increased from 23% in the sham-injured group to 55% to 63% in the 1-day and 7-day LFPI groups. Extracellular glucose concentrations during stimulation remained unchanged in the sham-injured and 7-day LFPI groups, but decreased 17% in the 1 day LFPI group. The extent of cortical degeneration around the stimulating electrode in the 1-day LFPI group nearly doubled when compared with controls. These results indicate that at 1 day after LFPI, the cortex can respond to stimulation with an increase in anaerobic glycolysis; however, this metabolic response to levels eliciting a vibrissa response via direct cortical stimulation appears to constitute a secondary injury in the TBI brain. PMID- 12902835 TI - Diffuse optical tomography of cerebral blood flow, oxygenation, and metabolism in rat during focal ischemia. AB - Diffuse optical tomography (DOT) is an attractive approach for evaluating stroke physiology. It provides hemodynamic and metabolic imaging with unique potential for continuous noninvasive bedside imaging in humans. To date there have been few quantitative spatial-temporal studies of stroke pathophysiology based on diffuse optical signatures. The authors report DOT images of hemodynamic and metabolic contrasts using a rat middle cerebral artery occlusion (MCAO) stroke model. This study used a novel DOT device that concurrently obtains coregistered images of relative cerebral blood volume (rCBV), tissue-averaged hemoglobin oxygen saturation (Sto(2)), and relative cerebral blood flow (rCBF). The authors demonstrate how these hemodynamic measures can be synthesized to calculate an index of the oxygen extraction fraction (OEF) and the cerebral metabolic rate of oxygen consumption (CMRo(2)). Temporary (60-minute) MCAO was performed on five rats. Ischemic changes, averaged over the 60 minutes of occlusion, were as follows: rCBF = 0.42 +/- 0.04, rCBV = 1.02 +/- 0.04, DeltaSto(2) = -11 +/- 2%, rOEF = 1.39 +/- 0.06 and rCMRo(2) = 0.59 +/- 0.07. Although rOEF increased in response to decreased blood flow, rCMRo(2) decreased. The sensitivity of this method of DOT analysis is discussed in terms of assumptions about baseline physiology, and the diffuse optical results are compared with positron emission tomography, magnetic resonance imaging, and histology observations in the literature. PMID- 12902837 TI - Evidence for a lactate pool in the rat brain that is not used as an energy supply under normoglycemic conditions. AB - Lactate derived from glucose can serve as an energy source in the brain. However, it is not certain how much lactate, directly taken from the blood circulation, may replace glucose as an energy source. This study aimed to estimate the uptake, release, and utilization of lactate entering the brain from the blood circulation. The change in cerebral venous-arterial glucose and lactate differences after lactate infusions in the anesthetized rat were measured. Ultrafiltration probes were placed in the aorta and in the jugular vein, and connected to a flow injection analysis system with biosensors for glucose and lactate. Measurements were taken every minute. Lactate efflux was observed at baseline, whereas an influx of lactate was seen during lactate infusion. Immediately after the infusion there was a net efflux of lactate from the brain. The results suggest that the majority of lactate moving into the brain is not used as an energy substrate, and that lactate does not replace glucose as an energy source. Instead, the authors propose the concept of a lactate pool in the brain that can be filled and emptied in accordance with the blood lactate concentration, but which is not used as an energy supply for cerebral metabolism. PMID- 12902836 TI - Functional brain mapping in freely moving rats during treadmill walking. AB - A dilemma in functional neuroimaging is that immobilization of the subject, necessary to avoid movement artifact, extinguishes all but the simplest behaviors. Recently, we developed an implantable microbolus infusion pump (MIP) that allows bolus injection of radiotracers by remote activation in freely moving, nontethered animals. The MIP is examined as a tool for brain mapping in rats during a locomotor task. Cerebral blood flow-related tissue radioactivity (CBF-TR) was measured using [14C]-iodoantipyrine with an indicator-fractionation method, followed by autoradiography. Rats exposed to walking on a treadmill, compared to quiescent controls, showed increases in CBF-TR in motor circuits (primary motor cortex, dorsolateral striatum, ventrolateral thalamus, midline cerebellum, copula pyramis, paramedian lobule), in primary somatosensory cortex mapping the forelimbs, hindlimbs and trunk, as well as in secondary visual cortex. These results support the use of implantable pumps as adjunct tools for functional neuroimaging of behaviors that cannot be elicited in restrained or tethered animals. PMID- 12902838 TI - Metabolic imaging of human cognition: an fMRI/1H-MRS study of brain lactate response to silent word generation. AB - Proton magnetic resonance spectroscopy (1H-MRS) allows in vivo assessment of the metabolism related to human brain functions. Visual, auditory, tactile, and motor stimuli induce a temporary increase in the brain lactate level, which may act as a rapid source of energy for the activated neurons. The authors studied the metabolism of the frontal lobes during cognitive stimulation and measured local lactate levels with standard 1H-MRS, after localizing the activated area by functional MRI. Lactate levels were monitored while the subjects either silently listed numbers (baseline) or performed a silent word-generation task (stimulus activation). The cognitive stimulus-activation produced a 50% increase in the brain lactate level in the left inferior frontal gyrus. The results show that metabolic imaging of neuronal activity related to cognition is possible using 1H MRS. PMID- 12902840 TI - Correlation between delayed neuronal cell death and selective decrease in phosphatidylinositol 4-kinase expression in the CA1 subfield of the hippocampus after transient forebrain ischemia. AB - Transient forebrain ischemia induces a delayed neuronal death in the CA1 area of the hippocampus. However, the mechanism leading to this phenomenon has yet to be established. The authors used an mRNA differential-display method to isolate genes for which mRNA levels change only in the hippocampus during ischemia/reperfusion. They succeeded in identifying the product of one down regulated gene as phosphatidylinositol 4-kinase (PI 4-K). Compared with control levels, PI 4-K mRNA expression in the hippocampus, but not the cerebral cortex, was significantly decreased by 30% and about 80% 1 and 7 days after ischemia/reperfusion, respectively. Interestingly, PI 4-K and PI bisphosphate levels were selectively decreased in the CA1 region, but not other regions, whereas TUNEL-positive cells could be detected 3 days after ischemia. Consistent with these results, PI 4-K expression was suppressed by hypoxia in SK-N-MC neuroblastoma cells before loss of cell viability. Overexpression of wild-type PI 4-K, but not the kinase-negative mutant of PI 4-K (K1789A), recovered the loss of viability induced by hypoxia. These findings strongly suggest that a prior decrease in PI 4-K and PI bisphosphate levels caused by brain ischemia/hypoxia is partly involved in delayed neuronal cell death. PMID- 12902839 TI - Induction of GRP78 by ischemic preconditioning reduces endoplasmic reticulum stress and prevents delayed neuronal cell death. AB - Although the endoplasmic reticulum (ER) is implicated in neuronal degeneration in some situations, its role in delayed neuronal cell death (DND) after ischemia remains uncertain. The authors speculated that ER stress is involved in DND, that it is reduced by ischemic preconditioning, and that ER stress reduction by preconditioning is due to ER molecular chaperone induction. The phosphorylation status of eukaryotic initiation factor 2alpha (eIF2alpha) and RNA-dependent protein kinase-like ER eIF2alpha kinase (PERK) was investigated in the rat hippocampus after ischemia with and without preconditioning. PERK is phosphorylated by ER stress, which phosphorylates eIF2alpha. To investigate the role of ER molecular chaperones in preconditioning, the authors examined GRP78 and GRP94 expression, both of which are ER chaperones that inhibit PERK phosphorylation, and compared their induction and ischemic tolerance time windows. Phosphorylation of eIF2alpha and PERK was confirmed after severe ischemia but was inhibited by preconditioning. After preconditioning, GRP78 was increased in the brain with a peak at 2 days, which corresponded with the ischemic tolerance time window. Immunoprecipitation and double staining demonstrated involvement of GRP78 in prevention of PERK phosphorylation. These results suggest that GRP78 induced by preconditioning may reduce ER stress and eventual DND after ischemia. PMID- 12902841 TI - Ultrasonographic assessment of global cerebral blood volume in healthy adults. AB - The authors describe a new ultrasonographic method for analysis of global cerebral blood volume (CBV) and its application under controlled hyperventilation. CBV was determined as the product of global cerebral blood flow volume (CBF) and global cerebral circulation time. CBF was measured by duplex sonography and calculated as the sum of flow volumes in both internal carotid arteries and vertebral arteries. Extracranial Doppler assessed cerebral circulation time by determining the time interval of echo-contrast bolus arrival between internal carotid artery and contralateral internal jugular vein. Forty four healthy volunteers (mean age 45 +/- 19 years, range 20-79 years) were studied. Mean CBV was 77 +/- 13 mL. CBV did not correlate with age, end-tidal carbon dioxide level, heart rate, or blood pressure. Hypocapnia was induced in 10 subjects by controlled hyperventilation. Mean reduction of end-tidal carbon dioxide values by 9 +/- 1 mm Hg led to a significant increase in cerebral circulation time (6.1 +/- 0.9 to 8.4 +/- 1.1 second, P < 0.0001) and a significant CBF decrease (742 +/- 85 to 526 +/- 77 mL/min, P < 0.0001), whereas CBV remained unchanged (75 +/- 6 to 73 +/- 10 mL). PMID- 12902842 TI - L-arginine levels in blood as a marker of nitric oxide-mediated brain damage in acute stroke: a clinical and experimental study. AB - There are no useful markers in blood of nitric oxide (NO)-mediated brain damage. Because l-arginine (l-arg) is the only known substrate for NO generation, the authors investigated the plasma profile of l-arg after cerebral ischemia, and the relationship of L-arg concentrations in blood with stroke outcome and infarct volume in a clinical and experimental study. l-Arg levels were determined with high-performance liquid chromatography in blood and CSF samples obtained on admission, and in blood 48 hours after inclusion, in 268 patients admitted with a hemispheric ischemic stroke lasting 8.2 +/- 5.9 hours. Infarct volume was measured by days 4 to 7 using computed tomography. Plasma l-arg profiles were analyzed in a separate group of 29 patients seen within 8 hours of onset (median, 4.5 hours) and in 24 male Fischer rats treated with subcutaneous vehicle or 20 mg/kg 1400W (a specific inducible NO synthase inhibitor) every 8 hours for 3 days after performing sham or permanent middle cerebral artery occlusion. Plasma l-arg concentrations decreased after the ischemic event, both in patients and rats, and peaked between 6 and 24 hours. In patients, there was a highly correlation between l-arg levels in CSF and plasma at 48 hours (r = 0.85, P<0.001). CSF and plasma l-arg concentrations negatively correlated with infarct volume (r = -0.40 and r = -0.35, respectively, P<0.001), and were significantly lower in patients with early neurologic deterioration and in those with poor outcome (Barthel index <85) at 90 days (P<0.001). In rats, the administration of 1400W resulted in a 55% significant reduction of infarct volume measured 72 hours after permanent middle cerebral artery occlusion, an effect that correlated with the inhibition caused by 1400W on the ischemia-induced decrease of plasma l-arg concentrations at 6 to 24 hours after the onset of the ischemia. Taken together, these data indicate that determination of l-arg levels in blood might be useful to evaluate the neurotoxic effects of NO generation. These findings might be helpful to guide future neuroprotective strategies in patients with ischemic stroke. PMID- 12902843 TI - Quantification of 5-HT2A receptors in the human brain using [18F]altanserin-PET and the bolus/infusion approach. AB - The aim of the present study is to describe and validate a method for accurate quantification of 5-hydroxytryptamine (5-HT)(2A) receptors using [18F]altanserin positron emission tomography (PET) and the bolus/infusion approach. A bolus/infusion ratio of 1.75 h aimed at attaining rapid steady state in blood and brain was predicted from previous bolus studies performed in our laboratory. The infusion schedule was tested in normal subjects (n = 10) using dynamic PET and frequent plasma sampling for 6 h. Steady state was attained in brain and plasma within 2 h, and time-activity curves remained constant for another 3 h. To represent free and nonspecifically bound [18F]altanserin and its radiolabeled metabolites only, cerebellum must show no displacement in 5-HT(2A) displacement studies. To validate this, saturating doses of cold ketanserin were administered and it was found that specific binding of [18F]altanserin decreased uniformly to the level of the cerebellum and no change in the cerebellar time-activity curve was found after ketanserin administration. A shorter experimental setup was tested in a second group (n = 20) including patients with neuropsychiatric disorders. Dynamic PET (five frames of 8 minutes each) and venous blood sampling at midscan time started 2 h after [18F]altanserin administration. The mean percentage rate of change per hour in the outcome parameter, DV(3)', was low (mean -0.3% h-1; range -7.3-7.2% h-1) and no correlation of DV(3)' versus time was demonstrated. It is concluded that 5-HT(2A) receptor studies can be conducted within 2 h of [18F]altanserin infusion, yielding reliable results. PMID- 12902845 TI - Expression of protease-activated receptor 2 in ulcerative colitis. AB - Although tryptase released from mast cells might play a key role in the pathogenesis of ulcerative colitis (UC), the role of protease-activated receptor 2 (PAR2), tryptase receptor, remains unclear in the pathogenesis of this disease. The expressions of PAR2 and tumor necrosis factor (TNF) alpha in nine UC tissues and nine normal tissues were examined by immunohistochemistry. TNF-alpha levels secreted from human leukemic mast cell line (HMC-1) after the treatment of PAR2 agonists were also measured by enzyme-linked immunosorbent assay. The PAR2 and TNF-alpha proteins were more significantly detectable in UC tissues than in normal tissues. Furthermore, 65.2% of PAR2+ cells and 66.4% of TNF-alpha+ cells in UC tissues were tryptase-positive cells. In other words, 60.6% and 46.3% of tryptase-positive cells in UC tissues were PAR2+ cells and TNF-alpha+ cells, respectively. A chi2 analysis showed correlation (p < 0.007) between PAR2 and TNF alpha in tryptase-positive mast cells. Moreover, PAR2 agonists significantly induced the TNF-alpha secretion from HMC-1. These results indicate that the activation of the mast cells through PAR2 may be involved in the pathogenesis of UC. PMID- 12902844 TI - Inducible and constitutive beta-defensins are differentially expressed in Crohn's disease and ulcerative colitis. AB - Antimicrobial peptides such as defensins provide nonspecific mucosal defense against a multitude of microorganisms. Recently, it has been shown that luminal bacteria may invade the mucosa in inflammatory bowel diseases, suggesting a defect in innate mucosal immunity. The aim of this study was to investigate the expression of human beta-defensins (HBD) in controls, Crohn's disease (CD), ulcerative colitis (UC), and unspecific inflammation. Up to 4 biopsies were taken from 103 patients (33 controls, 24 with Crohn's disease, 36 with ulcerative colitis, 10 with unspecific colitis). Mucosal mRNA was measured using real-time fluorescence temperature cycler reverse-transcription polymerase chain reaction with primers for HBD-1, HBD-2, HBD-3, tumor necrosis factor alpha, and interleukin 8. Mucosal HBD-1 expression was marginally decreased in both CD and UC. HBD-2 was increased exclusively in UC but not in CD. The expression of the novel defensin HBD-3 was strongly correlated with HBD-2 and also raised predominantly in UC. The expression of both inducible beta-defensins was enhanced in the state of inflammation. Expression of HBD-2 showed a weak correlation with interleukin 8 only in inflamed CD biopsies but not with tumor necrosis factor alpha. The missing induction of both inducible beta-defensins in CD as compared with UC may cause a defect in barrier function that predisposes to bacterial invasion. PMID- 12902846 TI - Celecoxib and rofecoxib potentiate chronic colitis and premalignant changes in interleukin 10 knockout mice. AB - Nonsteroidal anti-inflammatory drugs decrease sporadic colorectal carcinoma and adenomas in patients with familial adenomatous polyposis and in rodent models of sporadic colon cancer and familial adenomatous polyposis. Similarly, selective cyclooxygenase 2 inhibitors decrease adenomas in humans and rodents. However, their effects on chronic colitis and colitis-associated neoplasia are unknown. Interleukin 10-/- mice (C57/B6) were fed regular chow (n = 20) or chow with celecoxib (1,500 ppm, n = 18) or rofecoxib (75 ppm, n = 20) for 12 weeks. Twenty eight percent of the celecoxib group died versus 5% of the control and rofecoxib groups (p < 0.05 compared with control). Celecoxib and rofecoxib increased the incidence of colitis (26% vs. 92% and 68%, p < 0.01), colitis score (0.4 +/- 0.2 vs. 2.5 +/- 0.3 and 2 +/- 0.4, p < 0.01), aberrant crypt foci (0.5 +/- 0.3 vs. 3.7 +/- 2.6 and 2.8 +/- 0.7, p < 0.01), aberrant crypts per mouse (4.11 +/- 2.1 vs. 41.2 +/- 9.7 and 27.1 +/- 7.5, p < 0.01) and dysplasia (11% vs. 54% and 42%, p < 0.01). Similarly, indomethacin (9 ppm, n = 15) increased colitis score, aberrant crypt foci, and dysplasia after 27 days of treatment. Two selective cyclooxygenase 2 inhibitors exacerbate colitis and premalignant changes in the interleukin 10-/- mouse model of chronic colitis and colitis-associated colon carcinoma. PMID- 12902847 TI - Erythrocyte mean corpuscular volume as a surrogate marker for 6-thioguanine nucleotide concentration monitoring in patients with inflammatory bowel disease treated with azathioprine or 6-mercaptopurine. AB - Mean corpuscular volume may correlate with erythrocyte 6-thioguanine nucleotide concentrations in patients treated with azathioprine and 6-mercaptourine. We conducted a study of 166 patients with inflammatory bowel disease treated with azathioprine or 6-mercaptopurine to determine the relationship between mean corpuscular volume and erythrocyte 6-thioguanine nucleotide concentrations, disease activity as measured by the Inflammatory Bowel Disease Questionnaire (active disease <170, remission >170), and leukopenia. Blood was submitted for mean corpuscular volume, whole blood 6-thioguanine nucleotide concentration, and leukocyte count. The mean +/- SD mean corpuscular volume during treatment was 94.7 +/- 6.6 fL and the mean +/- SD change in mean corpuscular volume was 7.5 +/- 6.3 fL. There were significant correlations between mean corpuscular volume and erythrocyte 6-thioguanine nucleotide concentration (r(s) = 0.33, p < 0.001) and between change from baseline in mean corpuscular volume and erythrocyte 6 thioguanine nucleotide concentration (r(s) = 0.26, p = 0.001). There was no correlation between Inflammatory Bowel Disease Questionnaire scores and mean corpuscular volume values (r(s) = 0.01, p = 0.94). The mean corpuscular volume values in 55 patients with active disease and 111 patients in remission were similar (95.1 vs. 94.5 fL, p = 0.57). There was a weak negative correlation between the mean corpuscular volume and the leukocyte count, (r(s) = -0.18, p = 0.022). In patients with inflammatory bowel disease treated with azathioprine or 6-mercaptopurine, mean corpuscular volume and change from baseline in mean corpuscular volume correlated with erythrocyte 6-thioguanine nucleotide concentrations and negatively with leukocyte counts, but did not correlate with disease activity as measured by the Inflammatory Bowel Disease Questionnaire. Measurement of mean corpuscular volume is a simple and inexpensive alternative to measurement of 6-thioguanine nucleotide concentrations in patients treated with azathioprine or 6-mercaptopurine. PMID- 12902848 TI - Immune networks in animal models of inflammatory bowel disease. AB - The animal models of inflammatory bowel disease provide a framework to define the immunopathogenesis of intestinal inflammation. Studies in these models support the hypothesis that exaggerated immune responses to normal enteric microflora are involved in the initiation and perpetuation of chronic intestinal inflammation. A major pathway involves development of acquired immune responses by the interactions of CD4+ T-cell receptor alphabeta T cells with antigen-presenting cells (dendritic cells). Immunoregulatory cells, including Tr1 cells, Th3 cells, and CD4+ CD25+ T cells and B cells, directly or indirectly affect the T-cell receptor alphabeta T cell-induced immune responses and bridge innate and acquired immunity. The study of these complicated immune networks provides the rationale for the development of new therapeutic interventions in inflammatory bowel disease. PMID- 12902849 TI - Management of neoplastic polyps in inflammatory bowel disease. AB - Dysplasia-associated lesions or masses (DALMs) are a heterogeneous population of lesions with different endoscopic and morphologic features. Non-adenoma-like DALMs should be removed via colectomy. Adenoma-like DALMs that occur outside areas of colitis can be treated like sporadic adenomas and removed by polypectomy. Recent data suggest that adenoma-like DALMs located within areas of colitis should be removed by polypectomy with complete excision and multiple biopsies of the site. As long as there is no flat dysplasia or adenocarcinoma elsewhere in the colon, a colectomy is not required. However, this group of patients requires increased colonoscopic surveillance. PMID- 12902850 TI - Patients with low-grade dysplasia should be advised to undergo colectomy. PMID- 12902853 TI - Cutaneous nevi pigmentosus during infliximab therapy in a patient with Crohn's disease: fallacy or coincidence? PMID- 12902851 TI - Low-grade dysplasia: nonsurgical treatment. PMID- 12902854 TI - Proceedings of the RTOG (Radiation Therapy Oncology Group) Symposium: COX-2 Inhibitors and Cancer Therapeutics. June 2001. PMID- 12902855 TI - COX-2 inhibitors and cancer therapeutics: potential roles for inhibitors of COX-2 in combination with cytotoxic therapy: reports from a symposium held in conjunction with the Radiation Therapy Oncology Group June 2001 Meeting. AB - Tumor growth and angiogenesis are interdependent. Cyclooxygenase (COX) catalyzes the synthesis of prostaglandins (PGs) from arachidonic acid. Nonsteroidal antiinflammatory drugs (NSAIDs) inhibit COX-mediated synthesis of PGs. Although COX-1 is constitutively expressed in a wide range of tissues, COX-2 is cytokine inducible. Enhanced COX-2 expression has been attributed a key role in the development of inflammation and related processes observed in pathologically altered disease states. Two specific COX-2 inhibitors, namely, rofecoxib (Vioxx) and celecoxib (Celebrex), both oral agents and FDA approved, have been shown preclinically and clinically to have efficacy comparable to that of NSAIDs for relief of pain and inflammation in osteoarthritis, with decreased risk of gastrointestinal damage. Significant preclinical evidence strongly supports the potential role for these inhibitors for the treatment of cancer. On June 29, 2001, the Radiation Therapy Oncology Group (www.rtog.org), a National Cancer Institute-sponsored cooperative group, held a 1-day symposium focusing on the potential role of inhibitors of COX-2 in the treatment of cancer. This issue of the American Journal of Clinical Oncology contains the summary of those presentations. PMID- 12902856 TI - Development of COX inhibitors in cancer prevention and therapy. AB - On the strength of in vitro, in vivo, observational, and clinical data, nonsteroidal antiinflammatory drugs (NSAIDs)-also referred to as COX inhibitors have emerged as lead compounds for cancer prevention, and possible adjuncts to cancer therapy. Thus far, the routine use of NSAIDs for these indications is limited, largely owing to toxicity concerns, the paucity of efficacy data for any specific target organ, and uncertainties with regard to the most appropriate regimen (i.e., the best agent, formulation, dose, route of administration, and duration). Strategies to address these concerns primarily aim to improve the therapeutic index (i.e., benefit:risk ratio) of COX inhibitors by 1) minimizing systemic exposures whenever feasible, 2) achieving greater mechanistic specificity, 3) coadministering agents that provide prophylaxis against common toxicities, and 4) coadministering other effective anticancer agents. Clinical trials testing most of these strategies have been completed or are under way. The National Cancer Institute has a substantial research portfolio dedicated to the identification, testing, and development of NSAIDs as preventive and therapeutic anticancer agents. Discovering how to apply NSAIDs in persons with-or at risk for cancer, although challenging, has the potential for considerable clinical and public health benefits. PMID- 12902857 TI - COX-2 inhibitors as radiosensitizing agents for cancer therapy. AB - Prostaglandins have long been known to impact the radiosensitivity of cells and tissues, and many studies have centered on exploiting nonspecific prostaglandin inhibitors such as NSAIDs for therapeutic gain. These studies have ultimately been unsuccessful due to the lack of targeted specificity against the tumor. The discovery of the inducible cyclooxygenase enzyme (COX-2) and development of some highly selective inhibitors (which spare the constitutive COX-1 activity) has renewed excitement for modulating tumor prostaglandins as a method of specific radiosensitization of tumors, while sparing normal tissues. This review discusses these new data and generates a rationale for use of COX-2 inhibitors as radiosensitizing agents in cancer therapy. PMID- 12902858 TI - Rofecoxib (Vioxx), a specific cyclooxygenase-2 inhibitor, is chemopreventive in a mouse model of colon cancer. AB - The ApcDelta716 knockout mouse develops hundreds of intestinal polyps and smaller numbers in the colon because of the truncation of the suppressor protein Apc. We show inhibition of polyposis in the ApcDelta716 mouse by rofecoxib (Vioxx), a specific cyclooxygenase-2 (COX-2) inhibitor. Both the number and size of polyps in the ApcDelta716 mouse were markedly reduced by rofecoxib (Vioxx) treatment at plasma concentrations similar to those achieved in humans with antiinflammatory concentrations of Vioxx. Sulindac, a dual cyclooxygenase-1/2 inhibitor, also diminished size and number of polyps but to a lesser extent than rofecoxib. The protein expression of COX-1 or COX-2 was unchanged by treatment with rofecoxib or sulindac because these agents inhibit enzyme activity and prostaglandin product formation rather than transcription of the COX genes. The proangiogenic protein vascular derived endothelial growth factor was decreased in polyps treated with rofecoxib, whereas membrane-associated beta-catenin increased in rofecoxib treated polyps. DNA proliferation was decreased in polyps by both rofecoxib and sulindac treatment. Rofecoxib (Vioxx) is used clinically for osteoarthritis and pain, and in addition the results described here suggest that Vioxx may be useful as a chemopreventive in humans at risk for colorectal neoplasia. PMID- 12902859 TI - Cyclooxygenase-2 (COX-2) enzyme inhibitors and radiotherapy: preclinical basis. AB - Cyclooxygenase-2 (COX-2), an enzyme induced by proinflammatory cytokines, mitogenic substances, oncogenes, growth factors, and hypoxia, among others, is involved in the metabolic conversion of arachidonic acid to prostaglandins in inflamed tissues and neoplasia. COX-2 is often overexpressed in malignant tumors and premalignant lesions and is linked to carcinogenesis, maintenance of progressive tumor growth, and facilitation of metastatic spread. Because COX-2 may also be a determinant of tumor radioresistance, its inhibition or inhibition of its products (prostaglandins) may improve tumor response to radiotherapy. Preclinical studies have shown that treatment with selective COX-2 inhibitors significantly enhances tumor response to radiation without appreciably affecting normal tissue radioresponse. The underlying mechanisms of the COX-2 inhibitor radiation interactions seem to be multiple, with the enzyme inhibitor directly or indirectly augmenting tumor cell destruction by radiation. Thus, use of selective COX-2 inhibitors is a potential approach for improving cancer radiotherapy. PMID- 12902861 TI - Cyclooxygenase-2 (COX-2) and epidermal growth factor receptor (EGFR) expression in human pituitary macroadenomas. AB - Macroadenomas are tumors of the pituitary gland and are considered almost to be always benign and curable. The clinical manifestations of a pituitary tumor depend on the hormone secreted by the tumor as well as on the pattern of tumor growth within the sella turcica. Current trends attempt to target new molecular markers that also may serve as potential therapeutic targets. Cyclooxygenase-2 (COX-2) and epidermal growth factor receptor (EGFR) are upregulated in a number of epithelial tumors. No published reports exist about expression of COX-2 in pituitary macroadenomas, and only a few reports with differing results exist concerning EGFR expression in pituitary macroadenomas. This study sought to determine whether a relationship exists between COX-2 and EGFR expression and pituitary macroadenomas. Thirty specimens of pituitary macroadenomas were evaluated after being identified in the surgical pathology database of Thomas Jefferson University Hospital. The hematoxylin and eosin-stained slides were reviewed, and the representative paraffin blocks containing the index case were chosen and immunohistochemically stained for COX-2 and EGFR expression. The COX-2 and EGFR-stained slides were reviewed and an immunohistochemical score was calculated and analyzed. The pituitary macroadenomas were classified on the basis of hormone expression: none (nonsecreting), minor (nondominant, plurihormonal), single (dominant nonplurihormonal), or plurihormonal (dominant plurihormonal). The hormonal classification was then analyzed for association with COX-2 expression. COX-2 expression was significantly associated with plurihormonal pituitary macroadenomas (p value 0.03). COX-2 expression was significantly associated with expression of luteinizing hormone (p value 0.007) and with expression of thyroid-stimulating hormone (TSH) (p value 0.04). Additionally, COX 2 expression was significantly associated with single-hormone of pituitary adenoma (p value 0.049). The expression of COX-2 in 100% of the normal autopsy pituitary glands establishes an additional central nervous system location of COX 2 expression. EGFR was not expressed in any of the pituitary macroadenomas. The expression of COX-2 in plurihormonal pituitary macroadenomas, particularly those secreting TSH, may be a potential target for treatment in addition to surgical and/or radiotherapy treatment in these benign but clinically significant tumors. COX-2 is expressed in normal autopsy pituitary tissue. PMID- 12902860 TI - COX-2 inhibitor as a radiation enhancer: new strategies for the treatment of lung cancer. AB - Lung cancer is one of the most common causes of cancer-related mortality throughout the world, and the incidence continues to increase. Smoking is the number one cause of lung cancer. Emerging data have implicated cyclooxygenase-2 (COX-2) and prostanoid production in the pathogenesis of lung carcinoma. In invasive lung tumors, COX-2 upregulation has been reported in up to 90% of cases. COX-2 upregulation is an early event in the development of non-small-cell lung cancer and may be integral to the development of new blood vessels and production of specific proteases that are critical to growth and spread of lung malignancies. COX-2 inhibitors are known to enhance the chemosensitivity in COX-2 overexpressing lung cancer cell lines. Recently, we have demonstrated that selective COX-2 inhibitors also enhance the effect of radiation in COX-2 overexpressed cells. Therefore, inhibitors of COX-2 in combination with chemoradiation therapy may be an alternative strategy that can be tested in clinical trials. The combination of COX-2 inhibitors and radiation suggest a complementary strategy to target angiogenesis while potentially minimizing the impact on quality of life. Currently, several groups are conducting clinical trials in cervix cancer, lung cancer, and brain tumors, using inhibitors of COX-2 in combination with chemotherapy and radiation therapy. These clinical trials will help to elucidate the role of this interesting class. PMID- 12902862 TI - Initial experience combining cyclooxygenase-2 inhibition with chemoradiation for locally advanced pancreatic cancer. AB - Pancreatic cancer is a lethal disease that is resistant to chemotherapy and radiotherapy. Gemcitabine has recently been shown to be an improvement over 5 fluorouracil in patients with advanced disease. It is also a potent radiosensitizer, which has led to the investigation of gemcitabine with concurrent radiotherapy. However, preliminary results indicate that there are significant limitations to this approach in this challenging disease. Pancreatic cancer cells have alterations in many molecular signaling pathways that may be responsible for their resistance to cytotoxic therapy and aggressive behavior. Cyclooxygenase-2 (COX-2) is commonly overexpressed in pancreatic tumors, and preclinical evidence indicates that selective COX-2 inhibition enhances both chemotherapy and radiotherapy response, without affecting normal tissue damage. We have initiated preclinical studies as well as a phase I clinical protocol evaluating the combination of gemcitabine and celecoxib (Celebrex) with radiotherapy. In preclinical studies, celecelecoxib strongly enhanced the antitumor efficacy of chemoradiation. However, preliminary observations from both the preclinical experiments as well as the clinical protocol have revealed more toxicity with this combination than with gemcitabine and radiotherapy alone. These observations require further study, but are cause for concern when combining gemcitabine, radiotherapy, and celecoxib. PMID- 12902863 TI - Combination of a COX-2 inhibitor with radiotherapy or radiochemotherapy in the treatment of thoracic cancer. AB - Cyclooxygenase-2 (COX-2) is an enzyme involved in prostaglandin production in pathologic states such as inflammatory processes and cancer. The enzyme is often overexpressed in premalignant lesions and cancer, including cancers of the lung and esophagus. Inhibition of this enzyme with selective COX-2 inhibitors was found to enhance tumor response to radiation in preclinical studies, suggesting that these agents can improve the response of various cancers to radiotherapy. On the basis of these preclinical findings, clinical trials of the combination of celecoxib, a selective COX-2 inhibitor, with radiotherapy were initiated in patients with lung carcinoma and with chemoradiotherapy in patients with esophageal carcinoma. The rationale for using selective COX-2 inhibitors is discussed, and the current clinical protocols and the initial findings are described. PMID- 12902864 TI - Effect of the combination of docetaxel, zoledronic acid, and a COX-2 inhibitor on the growth of human breast cancer cell lines. AB - Current trends in the treatment of human tumors are with drug combinations that result in improved responses as well as the ability to use less toxic concentrations of the drugs. Recent reports have shown that COX-2 inhibitors and taxanes are effective in the suppression of human tumor growth. The bisphosphonate, zoledronic acid, primarily used in the treatment of bone metastases, also inhibits proliferation and induces apoptosis in human breast and prostate carcinoma and multiple myeloma. HER-2/neu overexpression has been suggested as a mechanism for resistance to both hormonal therapy as well as chemotherapy. This study examines the effects of combining a cyclooxygenase-2 inhibitor with zoledronic acid and/or docetaxel in a HER-2/neu transfected and control human breast cancer cell line. All three compounds produced dose dependent growth inhibition in both cell lines. The HER-2/neu transfected MCF/18 cells, however, were less sensitive to zoledronic acid than the control MCF/neo cells, 9% to 53% inhibition and 18% to 67%, respectively. Enhanced growth inhibition was observed in both cell lines with the combination of docetaxel and SC236 and the combination of docetaxel and zoledronic acid. The combination of SC236 with zoledronic acid also gave an enhanced inhibitory effect in the MCF/neo line. This combination, however, was additive in the HER-2/neu transfected MCF/18 cell line. The triple combination of SC236, zoledronic acid and/or docetaxel resulted in a small increase in growth inhibition compared to that seen with the double combinations. PMID- 12902865 TI - Cyclooxygenase-2 (COX-2) expression in human meningioma as a function of tumor grade. AB - Meningiomas are one of the most frequent central nervous system tumors, with an annual incidence in the United States of approximately 2.5 per 100,000 people. An intense interest exists in evaluating new molecular markers that may serve as potential therapeutic targets. Cyclooxygenase-2 (COX-2) is upregulated in a number of epithelial tumors, but to date, there are no published reports about the expression of COX-2 in meningioma. This study evaluated a possible relationship between COX-2 expression and malignant progression of meningioma. Eighty-three specimens of meningioma from the surgical pathology database of Thomas Jefferson University Hospital were evaluated. The hematoxylin and eosin (H&E)-stained slides were reviewed, and representative paraffin-embedded tissue sections containing the index cases were chosen and immunohistochemically stained for COX-2 expression. The H&E-stained sections of the individual tumors were classified according to the 1993 and 2000 World Health Organization (WHO) criteria for grading of meningiomas. The COX-2-stained slides were then reviewed and an immunohistochemical score was calculated and analyzed for statistical significance. The association between tumor grade and COX-2 expression is highly significant using the WHO-1993 grading criteria, (p = 0.012). Tumors with a more aggressive phenotype (benign --> atypical --> malignant) are associated with increasingly higher levels of COX-2. Using the 2000 WHO classification system, however, the association between tumor classification and COX-2 expression was not significant (p = 0.17), although the overwhelming percentage of tumors expressed COX-2. The association of COX-2 and meningioma is unique and represents a potential area for therapeutic intervention with selective COX-2 inhibitors, either as adjunct or in combination with radiation therapy. PMID- 12902867 TI - COX-2 inhibitors as radiation sensitizers for upper GI tract cancers: esophagus, stomach, and pancreas. AB - Cancers of the esophagus, stomach, and pancreas have been successfully treated recently with combinations of radiosensitizing chemotherapy and irradiation. New approaches building onto 5-fluorouracil chemoradiation include capecitabine (Xeloda) and irradiation. Capecitabine is an oral 5-fluorouracil (5-FU) prodrug that is more convenient than using infusional 5-FU, appears to have a similar therapeutic profile, and can be combined with daily irradiation. The addition of a cyclooxygenase-2 (COX-2) inhibitor is being investigated in upper gastrointestinal cancer sites because there is a high degree of overexpression of COX-2 in these cancers. PMID- 12902866 TI - Combination of radiation and celebrex (celecoxib) reduce mammary and lung tumor growth. AB - The selective cyclooxygenase (COX)-2 inhibitor, celecoxib, alone and in combination with radiation was investigated in vitro and in vivo. Murine mammary tumor line (MCa-35) and human lung carcinoma line (A549) have high and low basal levels of COX-2 protein, respectively. Treatment of both tumor cells with celecoxib alone resulted in a dose- and time-dependent reduction of cell number (clonogenic cell death) and tumor cell growth rate in vitro; however, inhibition of tumor cell growth by celecoxib was not correlated with the reduction of COX-2 protein in tumor cells. Although both tumor cell types had similar DNA damage after celecoxib treatment, significant induction of tumor cell apoptosis was only observed in MCa-35. Celecoxib-mediated radiation sensitization also occurred in MCa-35 cells determined by clonogenic assay, in part due to a G2/M arrest at 8 to 24 hours after treatment. The tumor growth inhibitory effects of celecoxib were also studied in vivo. It was found that celecoxib inhibited both tumor growth after intragastric administration of celecoxib (5 daily doses of 50 mg/kg). Combined with a single 30-Gy dose of radiation, celecoxib resulted in additive effects on A549 tumors. Celecoxib-treated A549 tumors had marginal reduction of total and perfused blood vessels compared with untreated controls. Reduction of tumor angiogenic cytokine and growth factor mRNA was associated with decreased perfused vessels. Finally, reduction of vascular endothelial growth factor protein after celecoxib was also observed in both tumor lines by Western blot. Our results indicate that the selective inhibition of COX-2 combined with radiation has potential application in radiotherapy, and celecoxib-mediated antitumor effects may act through different mechanisms including direct inhibition of tumor cell proliferation, alteration of tumor cell cycle, and antiangiogenesis. PMID- 12902868 TI - Celecoxib reduces skin damage after radiation: selective reduction of chemokine and receptor mRNA expression in irradiated skin but not in irradiated mammary tumor. AB - Inflammatory cytokine and chemokine production is mediated, at least in part, by prostaglandin E (PGE2). Cyclooxygenases, COX-1 and COX-2, are two key enzymes in the conversion of arachidonic acid to PGE2. Radiation induces the overproduction of cytokines and chemokines, and it also increases PGE2 production, both locally and systemically. In this study, we tested the effects of a COX-2 inhibitor (celecoxib) after 50 Gy radiation of MCa-35 tumor and cutaneous tissues of C3H/He mice. Preclinical toxicity endpoints and associated alterations in chemokine production and cellular infiltrates were measured. Celecoxib was given by daily gavage (50 mg/kg for 15 days), with the first dose delivered either 2 hours before, 2 days after, or 7 days after a single dose of radiation. Celecoxib treated animals had less inflammation of the dermis compared with saline-treated controls. Severe skin dermatitis occurred in 23.8% (5/21) of mice treated with 50 Gy, whereas only 17.6%, 5.3%, and 11.1% of mice in the 2-hour pre-, or the 2-day post-, and 7-day postirradiation groups, respectively, had severe dermatitis on day 20. The decreased skin toxicity scores were associated with a reduction of both blood vessels and focal necrosis in MCa-35 tumors. Celecoxib also significantly decreased C-C family chemokine (Rantes and MCP-1) mRNA expression in irradiated skin tissues, but not in tumor tissues, which was accompanied by a decrease in skin mRNA expression of both C-C (CCR2 and CCR5) and C-X-C (CXCR2 and CXCR4) chemokine receptors. A significant positive correlation was also found between skin damage (skin scores) and chemokine and its receptor mRNA expression in radiation-treated mice. Finally, celecoxib also decreased the infiltration of monocytes and neutrophils in locally irradiated tumor and surrounding normal tissue. The differential effects of celecoxib on inflammation help to explain the selective protection by celecoxib of irradiated cutaneous tissues without a concurrent protection of MCa-35 tumors. PMID- 12902869 TI - Why cyclooxygenase-2 inhibition plus chemotherapy? AB - New approaches to treating cancers are needed. Preclinical studies have identified numerous candidate genes/proteins that promote the cancer process. Cyclooxygenase-2 (COX-2) is a reasonable "target" because it is found in many epithelial tumors, has been shown to portend a poor prognosis, and is involved in many processes that promote cancer progression and chemotherapy resistance. Inhibition of COX-2 also has the potential to provide supportive care to patients with cancer. This article describes the rationale for performing a phase II trial of specific COX-2 inhibition in combination with chemotherapy to define toxicity and efficacy. However, as with most new therapies, phase III trials will be needed to determine whether specific COX-2 therapy is able to improve patient outcome with a reasonable safety profile. PMID- 12902871 TI - Development of aromatase inhibitors and their pharmacologic profile. AB - Major advances have been recently made in the treatment of postmenopausal women with hormone-sensitive breast cancer. It is widely accepted that estrogen is associated with the progression of hormone-sensitive breast cancer. Whereas previous therapy was based on surgical ablation (ovariectomy, adrenalectomy, and hypophysectomy) or additive treatment (estrogens, androgens), more recently antiestrogens (in premenopausal as well as postmenopausal women), medical ovarian ablation, and aromatase inhibition have replaced these procedures. The current focus of clinical research efforts is to maximize efficacy with these agents and determine an effective treatment strategy for these patients. This article reviews the development of aromatase inhibitors and inactivators (AIs), with particular emphasis on the potent third-generation agents now available. Also provided is information on the relative potency and efficiency of estrogen synthesis inhibition with these agents. Results from in vitro and in vivo studies of AIs suggest that there is variability in efficacy among agents in this class, and the potential clinical implications of these variations are discussed. PMID- 12902872 TI - A comparison of the efficacy of aromatase inhibitors in second-line treatment of metastatic breast cancer. AB - Randomized clinical trials have established the role of third-generation aromatase inhibitors (AIs) (letrozole, anastrozole, and exemestane) as standard treatment for patients with hormone-sensitive metastatic breast cancer who have experienced disease progression with antiestrogen therapy. Significant gains in clinical efficacy and improved tolerability over progestins (megestrol acetate) and the first-generation AI aminoglutethimide have positioned these agents above previous therapies. Estrogen receptor (ER) status remains the best predictive determinant of endocrine response, and further randomized trials with properly selected patient populations may distinguish individual AIs within this class. A recently completed, randomized, head-to-head phase III trial of letrozole versus anastrozole as second-line endocrine therapy demonstrated a significant difference in objective response rate for letrozole compared with anastrozole (19% versus 12%, respectively; P = 0.014), with similar time to progression. The improved efficacy and safety of AIs as second-line endocrine therapies has spawned trials of their use as first-line endocrine therapy versus tamoxifen for patients with metastatic breast cancer. Based on favorable results from these trials, letrozole and anastrozole have also been approved for use as first-line treatment of postmenopausal women with hormone receptor-positive metastatic breast cancer. PMID- 12902873 TI - Applicability of the intratumor aromatase preclinical model to predict clinical trial results with endocrine therapy. AB - Preclinical models and clinical studies have shown that aromatase inhibitors (AIs) are powerful inhibitors of estrogen synthesis and significantly suppress estrogen in vivo. For more than 20 years, standard first-line treatment for postmenopausal women with metastatic breast cancer has been the antiestrogen tamoxifen, a selective estrogen receptor modulator (SERM) with differential effects on breast, endometrial, bone, and vascular tissues. The estrogenic activity of tamoxifen is associated with deleterious clinical side effects, including vaginal bleeding, endometrial cancer, and thromboembolism. AIs are established second-line treatments in patients who progress with tamoxifen. Compared with progestins, such as megestrol acetate, or the earlier AIs aminoglutethimide and fadrozole, the new AIs, including exemestane, anastrozole, and letrozole, have increased efficacy and clinical benefit and cause fewer side effects in patients with metastatic breast cancer. Letrozole and anastrozole are approved first-line therapy for patients with metastatic breast cancer and as second-line treatment after tamoxifen failure. Studies in the intratumoral aromatase xenograft preclinical model have shown better responses with AIs than with antiestrogens in first-line therapy, and these data are consistent with the results from clinical trials. This model is now being used to assess whether combined or sequential administration of AIs with other agents may provide additional benefit. PMID- 12902874 TI - Emerging role of aromatase inhibitors in the adjuvant setting. AB - Aromatase inhibitors (AIs) have been approved as second-line treatment for estrogen receptor-positive (ER+) metastatic breast cancer after first-line treatment with the selective estrogen receptor modulator (SERM) tamoxifen. Anastrozole and letrozole have also recently been widely approved as first-line endocrine therapy for postmenopausal women with hormone receptor-positive metastatic breast cancer. The three third-generation selective oral AIs approved for use in the United States include two nonsteroidal agents, anastrozole (Arimidex) and letrozole (Femara), and the irreversible steroidal inhibitor exemestane (Aromasin). Several major ongoing clinical trials with a variety of treatment regimens are comparing the relative efficacy of tamoxifen with the steroidal and nonsteroidal AIs in the adjuvant setting. The first strategy compares an AI against tamoxifen directly. Among these are the ATAC (Arimidex, Tamoxifen Alone or in Combination) trial (anastrozole), the BIG FEMTA (Femara Tamoxifen Breast International Group) trial (letrozole), and the EXEM and TEAM (exemestane) trials. A second strategy is examining the use of an AI as an extension after the initial 5 years of tamoxifen. Examples of this trial design are the MA-17 (letrozole) and the National Surgical Adjuvant Breast and Bowel Project (NSABP B-33, exemestane) trials. A third approach is the use of these agents in sequence with tamoxifen as therapy within the initial 5 postoperative years. Examples of this approach are the International Collaboration Cancer Group trial (tamoxifen for 2-3 years followed by either tamoxifen or exemestane for the remainder of the 5-year period), the BIG FEMTA trial (patients are crossed over from tamoxifen to Ietrozole or letrozole to tamoxifen), and the Arimidex-Nolvadex (ARNO) trial (patients receiving tamoxifen are randomized either to continue with tamoxifen or to switch to anastrozole). A single trial is comparing tamoxifen and anastrozole as initial 5-year therapy, or a combination of the two. The study addressing this design is the ATAC trial. Finally, a small trial in Norway is comparing 2 years of an AI versus a placebo in very low-risk patients with receptor-positive breast tumors. Most adjuvant trials have companion studies associated with the main protocol. These are to determine the end-organ effects of the inhibitors and include measurements of quality of life, bone and lipid metabolism, and endometrial effects. This review addresses the clinical implications of these studies of AIs. PMID- 12902875 TI - Role of biologic markers in patient selection and application to disease prevention. AB - Aromatase inhibitors (AIs) are now under investigation for the treatment of early stage breast cancer and for disease prevention as alternatives to standard treatment with tamoxifen. Currently identified genetic risk factors of breast cancer include BRCA-1/BRCA-2 mutations, ATM mutations, and history of high estrogen levels, as evidenced by plasma analyses and/or dense bones. To date, estrogen receptor (ER) and progesterone receptor (PgR) status has predictive value for determining response to therapy in patients with hormone receptor positive breast cancer (ER+ and/or PgR+ tumors). Recent studies have shown AIs to be safer and more effective than tamoxifen in postmenopausal women with advanced disease. Some data suggest that letrozole may be a more effective treatment than tamoxifen for patients with ER+ and/or PgR+ early breast cancers expressing ErbB 1 and/or ErbB-2. Changes in cell proliferation markers (e.g., S-phase fraction and Ki67 antigen), plasma lipid levels, and the bone resorption marker C-terminal peptide are biomarkers that have been evaluated for preventive and prognostic value in breast cancer patients and normal volunteers. Results from biomarker screens can be used to define inclusion criteria for clinical trials and eventually to individualize treatment. Gene expression profiling (microarray analysis), i.e., genomic and proteomic studies, will probably advance the discovery of new biomarkers for breast cancer prevention and treatment. PMID- 12902876 TI - Examining quality of life issues in relation to endocrine therapy for breast cancer. AB - With the development of highly effective, well-tolerated third-generation aromatase inhibitors (AIs), these drugs will probably play an increasingly important role in all phases of breast cancer treatment. As a result, the impact of such hormonal agents on patients' quality of life bears rigorous investigation. In a randomized, multicenter, single-blind cross-over study, the AIs letrozole and anastrozole were evaluated for quality of life, toxicity, and patient preference. A total of 72 patients were enrolled and were treated with each drug for a 4-week period, with a 1-week drug-free washout period before cross-over to the alternate agent. Assessments included the FACT-ES, toxicity, and patient preference. The FACT-ES is a validated questionnaire designed to measure quality of life of women with breast cancer who are being treated with endocrine therapies. Letrozole was superior to anastrozole with respect to both quality of life and toxicity evaluations. In addition, at the conclusion of the trial, reduced nausea, hot flashes, and abdominal discomfort caused almost twice as many patients to prefer to continue with letrozole therapy than with anastrozole. Data from this recent trial indicate that letrozole is better tolerated and provides better quality of life than anastrozole for women with metastatic breast cancer. PMID- 12902878 TI - Low-dose radiosurgery for benign intracranial lesions. AB - This study assesses the efficacy and neurotoxicity of radiosurgical treatment of benign intracranial tumors using a linear accelerator, with relatively low dose and homogeneous dosimetry. Between June 1998 and July 2000, 27 patients were treated for benign lesions with radiosurgery using a 6-MV linear accelerator based X-knife system and circular collimators. The lesions included schwannoma, meningioma, papillary cyst adenoma, and hemangioblastoma. Five patients had tissue diagnosis. The mean peripheral dose to the tumor margin was 12.8 Gy. The mean dose to the isocenter was 16.3 Gy. One to five isocenters were used to treat these lesions, with a mean of 10 arcs per isocenter and mean collimator size of 1.25 cm. Follow-up information was available on all patients, with a mean follow up duration of 33 months. Six patients (22%) had improved symptoms and 21 (78%) had stable symptoms. Eight patients (30%) had regression of tumor and 19 had stable disease (70%). No patient had tumor progression, and Radiation Therapy Oncology Group (RTOG) grade III or IV toxicity did not occur in any patients. In 3 patients (11%), RTOG grade I or grade II neurotoxicity developed. Of these, one patient had worsening of a preexisting VIIth nerve deficit that required temporary oral methylprednisolone, and in two patients a mild trigeminal deficit developed that did not require any medical intervention. Low-dose homogeneous radiosurgery using a linear accelerator is an effective treatment for benign intracranial tumors. If lower, more homogeneous radiation doses produce responses as durable as higher doses, then toxicity might be further reduced. PMID- 12902879 TI - Role of Her-2/neu overexpression and clinical determinants of early mortality in glioblastoma multiforme. AB - Her-2/neu or c-erbB-2, a 185-kD protein is an important prognostic indicator/target for therapy in metastatic breast carcinoma. Recent reports have also identified a role for Her-2/neu overexpression in other solid tumors. We performed a retrospective analysis to evaluate the prevalence and prognostic role of Her-2/neu overexpression in patients with glioblastoma multiforme (GBM). Data collection (chart review) included demographic information, symptoms at presentation, histologic grade, survival time, and treatment offered. Testing for Her-2/neu overexpression was performed on paraffin-embedded archival tumor tissue using immunohistochemistry (IHC) (Hercep test). An IHC score of 2+ or greater was considered overexpression. An experienced pathologist who was blinded from the clinical history performed all the IHC testing. Between 1990 and 2001, 149 subjects (68 females, 81 males) with a biopsy-proven diagnosis of GBM were identified. Age range was 26 to 79 years (mean: 64 years) and overall mean survival was 12 months. Her-2/neu overexpression was detected in 23 patients (15.4%). Interestingly, the median survival for patients whose pathologic specimens revealed Her-2/neu overexpression was 4 months compared to those who lacked overexpression, in whom survival was 8 months. After adjusting for age, performance status, smoking history, and treatment, logistic regression analysis (with a survival of <3 months as the dependent variable) revealed that Her-2/neu overexpression significantly (p < 0.01) increased the odds of early mortality (<3 months). The results of our large study indicate that Her-2/neu overexpression may be a poor prognostic marker in patients with GBM. In addition, in a proportion of subjects (15.4%), Her-2/neu may be a potential target for tumor specific monoclonal antibody therapy. The role of trastuzumab (alone or in combination with conventional therapy) needs to be evaluated. PMID- 12902880 TI - A phase II study of the dolastatin 15 analogue LU 103793 in the treatment of advanced non-small-cell lung cancer. AB - A phase II study of the dolastatin 15 analog LU 103793 was conducted by the North Central Cancer Treatment Group in patients with advanced non-small-cell lung cancer (SCLC). Previously untreated patients received this agent at a dosage of 2.5 mg/m2 as a 5-minute intravenous infusion for 5 consecutive days every 3 weeks. Between September 1997 and July 1998, 17 patients were accrued in this study. Forty-two treatment cycles were administered with relatively modest toxicity. No responses were seen. This agent appears to be inactive in the treatment of advanced non-SCLC. PMID- 12902882 TI - Chemoembolization of hepatocellular carcinoma: results of a metaanalysis. AB - Transcatheter arterial chemoembolization is considered the mainstay of therapy for unresectable hepatocellular carcinoma. The purpose of this study was to assess the impact of such treatment on survival by performing a metaanalysis of all available randomized clinical trials comparing this form of therapy to supportive care. A MEDLARS search was conducted covering the years 1970 to 2002. Data analysis was performed according to methods described by Peto. The primary outcome of interest was the proportion of patients surviving 3 and 6 months after treatment. All analyses were performed on an intent-to-treat basis. A literature search yielded 1,100 citations, from which four met protocol-specified inclusion criteria. All studies contained an experimental and control arm totalling 268 patients. The odds ratio for 3- and 6-month survival were 1.31 (95% CI: 0.66 2.58) and 0.91 (95% CI: 0.49-1.68), which was not statistically significant. These data fail to show a survival advantage associated with therapeutic embolization versus supportive care alone in patients with unresectable hepatocellular carcinoma. Existing survival data from randomized controlled trials are of poor quality, and the paucity of patients in these trials eliminates the possibility of drawing meaningful conclusions regarding the effect of chemoembolization on patient survival from these studies. PMID- 12902881 TI - Phase II trial of infusional cyclophosphamide, idarubicin, and etoposide in poor prognosis non-Hodgkin's lymphoma. AB - The purpose of this study was to determine the complete response (CR) rate, failure-free survival (FFS), and overall survival (OS) of patients with poor prognosis intermediate-grade non-Hodgkin's lymphoma (NHL) after treatment with cyclophosphamide, idarubicin, and etoposide given as a continuous intravenous infusion (CIVI) over 96 hours (infusional CIE), including patients with relapsed/refractory disease and patients with no prior therapy but at least two poor-risk features by the age-adjusted International Prognostic Index. Forty-two patients with previously untreated NHL (N = 24) or relapsed/refractory (N = 18) NHL received cyclophosphamide (200 mg/m2/d), idarubicin (2.5-3.0 mg/m2/d) and etoposide (60 mg/m2/d) given by a 96-hour CIVI every 3 weeks for a maximum of 8 cycles. All patients also received granulocyte-colony-stimulating factor. CR occurred in 10 of 24 patients (42%; 95% confidence intervals [CI] 22%, 62%) treated with CIE as first-line therapy, and in 3 of 18 patients (17%; 95% CI 20%, 32%) treated with CIE as second-line or greater therapy. One-year FFS and OS were 42% and 64%, respectively, in patients with no prior therapy, and 17% and 56% in patients with prior therapy. Severe (grade III) or life-threatening (grade IV) toxicity included leukopenia (59%), anemia (61%), thrombocytopenia (31%), and infection (10%). Two patients (4%) died due to treatment related infectious complications. It is unlikely that infusional CIE produces a CR rate more than about 60% in poor-risk patients with intermediate-grade NHL when used as first line therapy, or more than about 30% in patients receiving the regimen as second line therapy. Substitution of idarubicin for doxorubicin in this setting, therefore, is not associated with an improved response rate. PMID- 12902883 TI - Patient characteristics and treatment factors affecting cosmesis following lumpectomy and breast irradiation. AB - Compared to a mastectomy, lumpectomy and breast irradiation provide an obvious cosmetic advantage. The majority of patients will have a good or excellent outcome with breast conservation therapy. However, poor or fair cosmetic results are reported in about 10% to 20% of patients. To evaluate factors influencing cosmetic outcome in women following lumpectomy and breast irradiation, all women (265) who presented for follow-up in a 6-month period after having previously been treated with postlumpectomy breast irradiation were thoroughly examined and assessed for cosmetic outcome factors including skin pigmentation changes, distortion of the breast, asymmetry, and differences in size between the two breasts. By multivariate analysis, skin pigmentation changes significantly correlated with axillary dissection (p = 0.0049) and black race (p = 0.001). An increased interval from surgery was associated with a decreased incidence of pigmentation changes (p = 0.0058). Smaller size of the treated breast was associated with a longer interval from surgery (p < 0.0001) and an increased separation between opposed tangential fields (p < 0.0001). One hundred twenty eight women (48.3%) had an excellent cosmetic result (no skin changes or deformity and very slight, if any, difference in size). An additional 110 patients (41.5%) had a good result (no skin changes and just a noticeable difference in size between the two breasts or skin changes with no difference in size between the two breasts). An excellent or good cosmetic result versus a fair or poor result was associated with white race (p = 0.0056), smaller separation between the tangential fields (p = 0.01), the use of a boost (p = 0.0025), and no use of tamoxifen (p = 0.025). PMID- 12902884 TI - Selection of patients with melanoma brain metastases for aggressive treatment. AB - The purpose of this study was to determine prognostic factors for patients with melanoma brain metastases that can be recommended for patient selection for clinical trials. A retrospective review was conducted of 65 patients irradiated for brain metastases from 1990 to 1997. Pretreatment factors analyzed for influence on survival included age, stage, Karnofsky Performance Status (KPS), extracranial metastases, the number and location of brain lesions, disease-free interval from initial diagnosis, total dose of radiation, and number of fractions administered. Prognosis was also analyzed by Radiation Therapy Oncology Group recursive partitioning analyses (RPA) classes. The data were analyzed using the Kaplan-Meier method. Median survival was 4 months. RPA class distribution was I 25%, II-48%, and III-28% with a median survival of 6.5, 3.5, and 2.5 months, respectively (p = 0.0098 by log-rank test). KPS less than 70% (p = 0.0039), and the presence of extracranial metastases (p = 0.03), predicted a worse prognosis on univariate analysis. Both factors remained significant on multivariate analysis. The prognosis of patients receiving radiotherapy for brain metastases is related to RPA class, the presence of extracranial metastases, and KPS. These criteria should be employed in selecting patients for aggressive protocol treatment, or for more protracted brain irradiation off protocol. PMID- 12902885 TI - Oral etoposide (VP16) in platinum-resistant epithelial ovarian cancer (EOC). AB - This phase II study evaluates the efficacy and toxicity of a prolonged schedule second-line and third-line treatment of oral VP16 in patients with measurable advanced ovarian cancer resistant to, or relapsed following, platinum-based chemotherapy. Twenty-two eligible women with progressive or relapsed ovarian cancer resistant to platinum-based therapy were included in this study. All the patients had received more than one prior treatment, and had evidence of disease progression within 6 months of the previous chemotherapy. Eleven patients had received more than two different chemotherapy regimens. Fifteen patients had received consolidation therapy with intraperitoneal cisplatin after an initial treatment course with six cycles of a platinum-based combination regimen. All patients with measurable disease observed in abdominal computed tomography scans were given oral VP16 at a daily dose of 50 mg/m2 for 14 consecutive days with 4 weekly intervals. Among 22 assessable patients, there were one complete response (CR) and three partial responses (PR), so the objective response rate, which is the addition of CR and PR rates, was 18%. Seven patients (32%) had stable disease. Median duration of response and stable disease was 2.5 months (range: 1 10 months). Overall median survival was 11 months from study entry (range: 3-36 months). Toxicity for most patients was mild, but a few severe myelotoxicities occurred, and there were no treatment-related deaths. According to World Health Organization toxicity criteria grade III/IV thrombocytopenia was seen in 4 of 22 patients, grade III/IV neutropenia in 6 of 22 patients, and grade III anemia was observed in 3 of 22 patients. Nonhematologic toxicity was mild, and mucositis was the most frequently observed nonhematologic toxicity. Oral etoposide has considerable activity with a tolerable toxicity profile for the treatment of platinum-resistant epithelial ovarian cancer. PMID- 12902886 TI - Triplet chemotherapy combination with gemcitabine, cisplatin and ifosfamide in patients with advanced non-small-cell lung cancer: phase II study. AB - A phase II study was conducted to evaluate the safety and efficacy of the combination GIP (gemcitabine, ifosfamide, and cisplatin) for the treatment of patients with advanced non-small-cell lung cancer (NSCLC). Thirty patients with stage III B/IV NSCLC were treated with a combination of GIP. Patients received gemcitabine 1,000 mg/m2 administered intravenously on days 1 and 8, ifosfamide 3,500 mg/m2 on day 2, and cisplatin 80 mg/m2 on day 2, repeated every 21 days. Two of the 30 patients (7%) showed a complete response and 14 patients (46%) showed a partial response. The overall response rate was 53%. The estimated median survival for all patients was 60 weeks. All patients enrolled onto the study were eligible for toxicity assessment. Toxicities were treatable and included World Health Organization grade III or IV leukopenia (29%), thrombocytopenia (18%), anemia (7%) and nausea, and vomiting (6%). Febrile neutropenia occurred in 3 of 30 patients. There were no treatment-related deaths. The combination therapy of GIP is active, well tolerated, and easy to administer on an outpatient basis in advanced NSCLC. PMID- 12902888 TI - Survival of patients with untreated cervical carcinoma. AB - Patients infrequently present with advanced-stage cervical malignancy in the United States, where access to care and aggressive screening, detection, and treatment regimens are the standard. Neglected cancer is more likely in countries with underdeveloped healthcare delivery systems; yet in this new millennium, we foresee not only an aging population with modern medicine able to prolong life expectancy, but also attentive screening regimens even amid the older old. Taking into account common comorbid illness from which patients used to die, informed consent in making a treatment in the robust and frail elderly becomes a new challenge. Recognizing an ever-growing population in today's culture of patient directed care, clinicians may find patients choosing alternative modalities or simply declining conventional therapies. The difficult dilemma then arises about how to inform a patient most accurately of her prognosis in an untreated condition. In the past, virtually all patients in the United States traditionally received some treatment for cervical cancer on diagnosis; thus, review was made of archival published literature to document survival of patients with untreated cervical cancer. PMID- 12902887 TI - Retrovesical soft-tissue metastasis of malignant thymoma: case report. AB - Malignant thymomas are usually confined to the mediastinum at the time of diagnosis and follow-up. Distant metastasis is distinctly rare. This is the first clinical case report of a thoracic malignant thymoma with distant soft-tissue metastasis that involves the retrovesical area to include the seminal vesicle. PMID- 12902889 TI - Pulmonary metastases tumor doubling time: assessment by computed tomography. AB - Growth rate is a prognostic factor for pulmonary metastases treatment. We used computed tomography (CT) to quantify tumor doubling time (Tdt) variation of pulmonary metastases in the same patient, from different histologies and sizes. We prospectively evaluated chest CTs of patients with pulmonary metastases under no cancer treatment. Tdt of 408 pulmonary nodules in 21 patients was determined. One hundred twenty-three (30%) remained unaltered, 44 (11%) had reduction, and 241 (59%) increased in size. Nodules that grew presented a mean Tdt of 118 +/- 134 days (range: 22-930 days). There was high intraindividual variability of mean Tdt with no difference between histologies. Smaller nodules presented greater tendency to grow, and lower Tdt. There is large variation in clinical behavior for pulmonary metastases in the same patient. Each nodule should be evaluated independently. Selection of a single nodule to evaluate metastatic disease behavior could influence the therapeutic decision. PMID- 12902890 TI - Docetaxel and vinorelbine in recurrent head and neck cancer: pharmacokinetic and clinical results. AB - The purpose of this study was to evaluate pharmacokinetic parameters, efficacy, and toxicity of a combination of docetaxel (DTX) and vinorelbine (VNB) in recurrent heavily pretreated squamous cell head and neck cancer. Twenty-nine patients previously treated with concomitant chemoradiotherapy (n = 14), surgery plus radiotherapy (n = 13), surgery+concomitant chemoradiotherapy (n = 1) and radiotherapy alone (n = 1) were enrolled; 9 patients had received 1 or more courses of palliative chemotherapy. Twenty-one patients had a local-regional recurrence, and 8 patients had metastases. The doses were 80 mg/m2 for DTX and 20 mg/m2 for VNB on day 1 every 21 days for a maximum of 6 cycles. Pharmacokinetic evaluations were performed on 24 patients; in a group of 12 patients, VNB administration immediately followed DTX infusion (schedule A), and in 12 patients VNB administration was immediately followed by DTX infusion (schedule B). Twenty nine patients received a total of 137 cycles (median per patient, 5). Neutropenia was the most frequent and severe side effect (grade IV in 79%; grade III in 21%). Grade IV (7%) and III (14%) infections were observed in the first 12 patients; ciprofloxacin prophylaxis in the following 17 patients reduced the severe toxicity to 0%. The overall response rate was 49%, which included 3 of 29 complete responses (10%) and 11 of 29 partial responses (38%). Median complete and partial response durations were 20+ and 5.5 months, respectively. Overall median survival was 10 months (range, 2-30+). The mean values of area under the curve, mean residence time (MRT), and C(max) of VNB were significantly lower for schedule A than for schedule B. The mean values of VNB clearance were significantly higher for schedule A than for schedule B. Neutrophil count at the nadir was much lower for patients receiving schedule B. The DTX-VNB combination is effective in heavily pretreated patients with a short-lasting manageable toxicity. Pharmacokinetic evaluations suggested that the sequence DTX --> VNB is safer than the sequence VNB --> DTX. PMID- 12902891 TI - Impact of hormone therapy when combined with external beam radiotherapy for early stage, intermediate-, or high-risk prostate cancer. AB - The purpose of this investigation was to explore the potential benefit of hormone therapy in addition to external beam radiotherapy for patients with early-stage (T1-2), intermediate-(prostate-specific antigen [PSA] > 10 or Gleason score >or= 7) or high-risk (PSA > 10 and Gleason score >or= 7) prostate cancer. The charts of 412 patients with early-stage intermediate- and high-risk prostate cancer treated with external beam radiotherapy with or without a 4-month total androgen blockade were reviewed. The groups were balanced with respect to age, pretreatment PSA, and stage, but differed with respect to Gleason score and radiation dose. Biochemical failure rates, as defined by the ASTRO consensus panel, were compared between those receiving and those not receiving hormones. With a median follow-up of 2.0 years, the biochemical failure rate was 12.1 versus 23.1% (p = 0.02) in favor of those receiving hormones. This difference was seen for the subgroups followed for more than 6 months (12.5 vs. 25.0%), more than 9 months (14.5 vs. 26.3%), and more than 12 months (17.3 vs. 27.0%). Thus, biochemical failure decreased with the administration of hormone therapy in this group of patients with early stage, intermediate- or high-risk prostate cancer. This finding requires validation by ongoing randomized trials. PMID- 12902892 TI - Genetic alterations and MSI status in primary, synchronous, and metachronous tumors in a family with hereditary nonpolyposis colorectal cancer (HNPCC). AB - In colorectal cancer, different levels of microsatellite instability (MSI) have been described: high-frequency MSI, low-frequency MSI, and stable microsatellites. MSI-H characterizes a unique clinical and pathologic phenotype known as hereditary nonpolyposis colorectal cancer syndrome (HNPCC). In this case, an increased incidence of synchronous and metachronous tumors has been reported, but there are few reports with standardized criteria of MSI in HNPCC associated tumors. The authors attempted to establish whether tumors of the HNPCC spectrum with different levels of MSI could predict the development of metachronous carcinomas. We have examined the levels of MSI at loci frequently affected in colorectal cancers in primary, synchronous, and metachronous tumors in a family that fulfils the Amsterdam criteria for HNPCC. This family presents colorectal cancers, HNPCC-extracolonic tumors (endometrial and ureter), and tumors (breast and bladder) not described in the HNPCC spectrum. The tumors exhibited MSI-H, irrespective of their location and regardless whether they were primary, synchronous, or metachronous, with the only exception of both endometrial tumors that showed low-frequency MSI tumors (MSI-L). Our results suggest that not only colorectal tumors with MSI-H result in a potential marker for the determination of high-risk individuals for metachronous and synchronous tumors, but also MSI-L endometrial tumors might be considered as indicative of high-risk individuals. PMID- 12902894 TI - Adjuvant treatment in elderly patients with breast cancer: critical review of clinical practice. AB - Today the use of adjuvant treatment of breast cancer is unquestionable in the management of this disease. Both chemotherapy and hormonal therapy have proved to be beneficial, not only with respect to the reduction of the risk of recurrence, but also with respect to mortality. However, in elderly patients, this therapeutic approach is occasionally the subject of controversy, due to the undervaluing of the tumoral disease with respect to the multiple pathology frequently present in these patients. This study analyses a retrospective series of 100 patients more than 70 years old with breast cancer who underwent radical surgery between 1990 and 1998, with an extension study without evidence of metastasis and a minimum follow-up of 2 years. As occurs in the population of this age, in our series 77% of the patients presented with concomitant disease under medical treatment, and although the majority received adjuvant treatment with tamoxifen, the principal cause of death in this series was the breast cancer that had been diagnosed. PMID- 12902893 TI - Multivariate analysis of treatment outcome in patients with esophageal carcinoma treated with definitive radiotherapy. AB - To evaluate patient characteristics and treatment factors influencing outcome of patients treated with definitive radiotherapy, we performed retrospective analysis. From 1983 to 2000, 154 patients who were diagnosed as esophageal carcinoma without distant metastasis received definitive radiotherapy with (N = 90) or without (N = 64) systemic chemotherapy. One hundred forty-two males and 12 females were entered in the analysis. Thirty-four patients received an additional boost of intracavitary brachytherapy (ICBT). The median patient age was 68 years (range: 46-86). Disease stage was distributed as stage I, II, III, and IV for 33, 42, 33, and 45 patients, respectively. External beam radiotherapy was prescribed with a median 63 Gy (range: 38-77.8 Gy). The 2- and 5-year overall survival (OAS) and local control (LC) rates were 40.8/18.4% and 48.6/28.9%, respectively. In uni /multivariate analyses, significant prognostic factors of OAS proved to be advanced T stage, absence of ICBT, and age less than 65 years. As for LC, adverse prognostic factors of uni/multivariate analysis were advanced T stage and poor performance status. The pretreatment T stage showed the most powerful influence on both survival and LC. Combination use of ICBT is proven to refine treatment outcome, although eligible criteria should be decided by a prospective study. PMID- 12902895 TI - Impact of intravesical chemotherapy versus BCG immunotherapy on recurrence of superficial transitional cell carcinoma of the bladder: metaanalytic reevaluation. AB - Bacille Calmette-Guerin (BCG) immunotherapy is currently considered the most effective agent in the management of superficial bladder cancer. Prior work suggests that the efficacy of intravesical chemotherapy in preventing tumor recurrence may be greater than previously suggested. This latter finding, therefore, brings into question the currently perceived superiority of BCG treatment for this disease. A metaanalysis was performed to rigorously examine existing data relevant to this relationship and to quantify the relative efficacy of both treatment modalities on tumor recurrence. A prospective protocol outlining the above-noted metaanalysis was initially developed followed by a thorough search of the existing published literature using strict eligibility criteria. Nine randomized trials were found that met protocol specifications. These reports contained data on 2,261 patients that were statistically combined using a fixed effects model (Peto). The outcome of interest was the proportion of patients with recurrence at 1, 2, and 3 years following intravesical therapy (i.e., a summary odds ratio, ORp). Combining all nine trials using 1-year recurrence as the endpoint demonstrated significant statistical heterogeneity, although the ORp favored BCG over intravesical chemotherapy (0.89 [0.74-1.07]). This precluded statistical pooling of the data and sensitivity analyses were performed to determine the source of heterogeneity. These tests showed that the prior chemotherapy treatment in a large number of the randomized trials biased study results in favor of the BCG arms. Once the data were stratified on presence or absence of prior drug treatment, intravesical chemotherapy reduced 1-, 2-, and 3-year recurrence by 21% to 82% versus BCG, depending on the endpoint of interest. The available data suggest that clinical trials directly comparing intravesical BCG to intravesical chemotherapy must stratify on the presence or absence of prior chemotherapy. Recurrences following prior intravesical chemotherapy appear less responsive to drug therapy than those in chemotherapy naive patients. The currently perceived superiority of BCG therapy may therefore be an artifact of this phenomenon, since most randomized trials include chemotherapy failures in their chemotherapy treatment arms. PMID- 12902896 TI - Successful treatment of refractory low grade duodenal lymphoma with rituximab, an anti-CD20 monoclonal antibody. AB - Duodenal lymphoma accounts for a small percentage of gastrointestinal tumors and has a very poor prognosis. The majority of patients have little or no cure with limited time of remission. Median survival is 5 to 10 years, and most patients die of lymphoma, its complications, or complications of therapy. We report a patient with lymphoma refractory to treatment with cyclophosphamide, vincristine, and prednisone, who was successfully treated with rituximab, a CD-20 monoclonal antibody. PMID- 12902897 TI - Prognostic factors in localized soft-tissue sarcomas. AB - The prognostic factors associated with local failure and overall survival and the effect of radiotherapy were determined in 77 patients with localized (extremity and nonextremity) operable soft-tissue sarcoma. There were 52 male and 25 female patients; median age was 50 years (range: 15-83). Histologic grade of the tumors was as follows: low-intermediate grade in 32 cases and high grade in 29 cases. The primary tumors were treated by marginal resection (20 patients), wide resection (52 patients), and radical resection (5 patients). Adjuvant radiotherapy was applied to 50 (65%) patients. The 5-year local recurrence-free survival rate was 70.6%. Treatment with adjuvant radiotherapy and development of metastases were the significant prognostic factors associated with local recurrence. Radiotherapy was more effective in patients with tumors 10 cm or larger, marginally resected, extremity located, and high grades. The overall survival rate was 64.4% at 5 years. Significant adverse prognostic factors were high grade tumors, presence of local recurrence, and development of metastases in univariate analyses. Development of metastases and old age were the only adverse prognostic factors by multivariate analysis. The best 5-year survival rate was obtained in female patients younger than 50 years (90%). The present study demonstrated the importance of adjuvant radiotherapy and development of metastases as prognostic factors for local control. Again, development of metastases and age were the most important prognostic factors in operable soft tissue sarcomas. PMID- 12902898 TI - Prognostic value of serum IL-18 and nitric oxide activity in breast cancer patients at operable stage. AB - Interleukin-18 (IL-18) is a multifunctional cytokine that was previously termed interferon-gamma-inducing factor. It has been suggested that serum IL-18 level may be used as a prognostic factor in some cancer types. Nitric oxide is a potent biologic molecule involved in the pathogenesis of cancer. In this study, we measured serum IL-18 and nitrate + nitrite levels in 56 patients with nonmetastatic breast cancer and 14 control subjects. Serum IL-18* and nitrate + nitrite** levels were significantly higher in patients with breast cancer when compared to the control subjects (*p < 0.05, **p < 0.001). Serum IL-18 levels were significantly higher in patients whose tumor size was greater than or equal to 5 cm when compared to patients whose tumor size was less than or equal to 2 cm (p < 0.05). Patients who were axillary lymph node negative (ALN) had lower serum IL-18 levels when compared to patients with positive ALN (p < 0.001). Serum IL-18 levels were significantly higher in patients with stage IIB or IIIA when compared to patients with stage I or IIA (p < 0.05). There was no significant difference in serum nitrate + nitrite levels in terms of age, tumor stage, estrogen receptor, and menopausal and ALN status (p > 0.05). In conclusion, serum IL-18 level may be a useful marker to predict prognosis of patients with breast cancer in complete remission after surgery. Long-term follow-up is required to clarify this hypothesis. PMID- 12902900 TI - Melphalan for the treatment of patients with recurrent epithelial ovarian cancer. AB - The purpose of this study was to determine the response to melphalan in patients with recurrent epithelial ovarian cancer after platinum-based therapy. This retrospective observational study analyzed 10 patients with recurrent epithelial ovarian carcinoma treated with melphalan between August 1995 and April 2001. All had received primary platinum-based therapy. Nine of the 10 patients had chemosensitive disease. All but one patient had received one or more second-line therapies prior to melphalan. The median time to recurrence after first-line therapy was 26 months (range, 3-68). Treatment with melphalan resulted in 2 (20%) complete responses and 1 (10%) partial response (response rate, 30%; 95% CI 8%, 65%). The median progression-free interval after initiation of melphalan therapy was 8 months (range, 3-23). The most common side effects were grade I thrombocytopenia (20% of courses) and grade II leukopenia (18% of courses). The use of melphalan as palliative chemotherapy in patients with recurrent ovarian cancer results in response rates similar to those reported with other more expensive agents. Melphalan at the doses reported here has a favorable toxicity profile. PMID- 12902899 TI - Cholangiocarcinoma: the impact of tumor location and treatment strategy on outcome. AB - The purpose of this study was to evaluate how the outcome of patients with extrahepatic cholangiocarcinoma (EHBC) may have been influenced by tumor location and treatment selection. The primary endpoint of this study is overall survival (OS). Between January 1983 and December 1997, 221 patients with biliary tumors were evaluated at Thomas Jefferson University Hospital. Of these, 118 fit the inclusion criteria for this study. The extent of disease was assessed by computed tomography, percutaneous transhepatic cholangiography or endoscopic retrograde cholangiopancreatography, magnetic resonance imaging, and ultrasonography. All patients had histologic confirmation of malignancy. Roux-en Y, hepaticojejunostomy, or choledochojejunostomy followed surgical resection of the primary tumor. Palliative measure (PS) included biliary catheter placement without brachytherapy or external beam irradiation (RT). RT was delivered via high-energy photons. Intraluminal brachytherapy was performed via percutaneous biliary catheterization with iridium-192 ribbon sources. Chemotherapy consisted of either intravenous 5-fluorouracil alone or in combination with doxorubicin, mitomycin C, or paclitaxel. PS consisted of metal bile duct stent placement. Median follow-up time for the entire group was 102 months and 43 months for patients who were still alive at the conclusion of the study period. Patients with proximal tumors underwent resection (n = 5), surgery and RT (n = 23), RT only (n = 31), chemotherapy only (n = 6), or PS (n = 12). Patients with distal tumors were treated with surgical resection (n = 17) or a combination of surgery and RT (n = 13), RT only (n = 6), or PS (n = 4). Median survival time (MST) for all 118 patients was 22 months. The MST for patients with distal tumors was 47 months versus 17 months for those with proximal tumors. The MST has not been reached for patients with distal EHBC treated with surgical resection and postoperative RT, whereas the median survival for those treated with surgery alone is 62.5 months. However, 4 of 17 of these patients had in situ carcinoma. Six patients had distal tumors treated with RT only with a MST of 6 months. Patients with proximal tumors treated with surgery and RT had a superior OS at 5 years compared to patients treated with RT alone (24 vs. 13 months; p = 0.007). There was an improved OS in patients with proximal tumors treated with surgical resection and RT compared to surgery alone (p = 0.023). There is no discernable influence of chemotherapy on outcome in patients with proximal EHBC. The MST for patients treated with PS was 3.5 months. Surgery and postoperative RT appear to be better than either surgery or RT alone in patients with proximal EHBC. In patients with distal EHBC, the addition of resection and RT appears to offer an advantage, which is increasingly apparent with longer follow-up time. The prognosis remains dismal for patients treated with palliative intent. PMID- 12902901 TI - Inducible nitric oxide synthase and nitrotyrosine in mice with radiation-induced lung damage. AB - The purpose of this study was to determine if radiation-induced lung damage is associated with induction of nitric oxide synthase (NOS) II and nitrotyrosine in an irradiated lung mouse model. The thorax of BALBc mice were exposed to 14 Gy radiation (experimental) or no radiation (control) and killed after at 1, 3, 6, 12, and 24 hours; 3, 15, and 30 days; and 3 and 6 months after treatment. Lung sections were processed for immunohistochemistry using NOS II and nitrotyrosine polyclonal antisera and in situ hybridization using 35S labeled probes for mouse NOS II. Quantitative analysis of experimental and control sections showed significant induction of NOS II and nitrotyrosine in alveolar macrophages from 6 hours to 30 days postirradiation, which was diminished by 3 months. The airway and alveolar epithelium and vascular endothelium showed strong NOS II expression at 15 to 30 days postirradiation. Nitrotyrosine immunostaining was also strongly evident in the alveolar epithelium and vascular endothelium during this period. There was little or no NOS II or nitrotyrosine in the sham control lungs throughout the study. These findings demonstrate increased formation of both NO and nitrotyrosine after radiation treatment and suggest a role for these molecules in the pathogenesis of radiation-induced lung damage. PMID- 12902902 TI - Radiosensitivity in multidrug-resistant and cisplatin-resistant human carcinoma cell lines. AB - The radiosensitivity of a multidrug-resistant (MDR) clone and a cisplatin resistant clone was compared with that of their parental chemosensitive cell lines. The LoVo cell line was derived from a human colon carcinoma, and LoVo-R was the MDR clone. The MDR phenotype is attributable to an increased drug efflux mediated by the P-glycoprotein and involves several classes of structurally unrelated drugs. The 2008 cell line was derived from a human ovary carcinoma and C13 was the cisplatin-resistant clone. Reduced cisplatin accumulation and elevated plasma membrane potential partially account for the drug resistance of C13 cells. The chemoresistance of LoVo-R and C13 cells was confirmed by cytotoxicity tests consisting of 24-hour paclitaxel and 1-hour cisplatin incubation, respectively. The radiosensitivity was evaluated by a clonogenic test. The dose-reducing cell survival fraction from 1 to 0.37 (D(0)), the quasi threshold dose (Dq), and the survival fraction (SF) after 2 or 4 Gy were determined for each cell line. D(0), Dq, and SF(2) were 1.3 +/- 0.4 Gy, 2.1 +/- 0.6 Gy, and 43 +/- 4% for the LoVo cell line and 1.0 +/- 0.2 Gy, 1.7 +/- 0.4 Gy, and 45 +/- 8%, respectively, for the LoVo-R cell line. D(0), Dq, and SF(4) were 1.7 +/- 0.3 Gy, 3.1 +/- 0.4 Gy, and 43 +/- 12% for 2008 cells and 2.6 +/- 0.5 Gy, 4.3 +/- 0.6 Gy, and 53 +/- 11%, respectively for C13 cells. No significant differences were found between LoVo and LoVo-R cells, whereas C13 cells showed a significantly greater D(0,) Dq, and SF(4) than 2008 cells (p <0.05). Incubation of 2008 and C13 cells with subcytotoxic buthionine (BSO) before and after irradiation partially restored C13 radiosensitivity. In fact, D(0) dropped from 2.8 +/- 0.1 to 2.0 +/- 0.3 Gy in C13 cells with and without BSO, whereas it was 1.9 +/- 0.2 Gy in 2008 cells in the absence and presence of BSO. The total glutathione content (GSH) of C13 cells was 1.5-fold higher than that of 2008 cells. BSO treatment caused a partial depletion of GSH in 2008 and C13 cells, but their radiosensitivity did not change accordingly. PMID- 12902903 TI - Expression of Ku80 in cervical cancer correlates with response to radiotherapy and survival. AB - To reveal the genes relevant for prediction of cervical cancer after radiotherapy, we previously carried out cDNA microarray experiments on primary cervical cancer comparing patients with a complete response (CR) and those with no change (NC). Some of these genes had already been associated with the radiation response, such as x-ray repair cross-complementing 5 (XRCC5), which was found more in radioresistant tumors than in radiosensitive ones. The aim of this study was to confirm the possible roles of XRCC5 mRNA levels by a real-time polymerase chain reaction method in 20 cervical cancers, and Ku80 protein, which is the gene product of XRCC5, using a histopathologic method of formalin-fixed sections of tumor biopsies in determining tumor response to radiotherapy and survival in 89 patients with cervical cancer. The levels of XRCC5 mRNA were 10(4.82) +/- 10(0.2) copies/microg total RNA in tumor tissues in the CR group (mean +/- standard deviation) and 10(4.95) +/- 10(0.32) copies/microg total RNA in those in the NC group. The levels of XRCC5 mRNA were not significantly different between the CR and NC groups. Histopathologic methods revealed 29.2% (26 of 89) of the patients to be Ku80-negative, with Ku80-positive findings in 70.8% (63 of 89). Of the Ku80-negative patients, 19 had CR, 3 had a partial response (PR), and 4 had NC. Of the Ku80-positive patients, 25 had CR, 22 had PR, and 16 had NC. Ku80-negative tumors showed significantly better responses than Ku80-positive ones, comparing CR and PR/NC responses (p = 0.01). In addition, overall survival was significantly better in the Ku80-negative patients as compared with those who were Ku80-positive (p = 0.04). The results of this study suggest that a low expression of Ku80 protein leads to radiosensitivity in cervical cancer and that Ku80 might play a role in treatment outcome. PMID- 12902904 TI - A simple method of producing low oxygen conditions with oxyrase for cultured cells exposed to radiation and tirapazamine. AB - Several methods of establishing low O(2) conditions have been used in studies on the response of cultured cells to radiation and other agents. These methods, eg, gassing culture vessels with O(2)-free nitrogen with or without carbon dioxide or placing high cell-density suspensions in sealed glass ampoules to consume O(2) in the ampules, can be technically demanding and have experimental limitations. We introduce a simple, versatile, and reliable method of producing low O(2) conditions without special equipment or changes in culture conditions unrelated to hypoxia. The method is based on the ability of Oxyrase (Oxyrase, Inc., Mansfield, OH), membrane fragments prepared from Enterococcus coli, to consume O(2) in solution and is confirmed in the present study by 2 analytical methods. The effects of low O(2) conditions induced by Oxyrase on cellular responses to radiation and treatment with the bioreductive agent tirapazamine (TPZ) were examined with Chinese hamster V79 and human glioma U373 cells. Measured by clonogenic and MTT assays, these cells were less sensitive to radiation but more sensitive to TPZ in treatment media containing native Oxyrase than in media containing heat-inactivated Oxyrase. In addition, Oxyrase treatment increased the basal activity of mitogen-activated protein kinase (ERK1/2) but suppressed its activation induced by radiation. The results suggest that this method might also be useful for other in vitro cancer biologic investigations requiring a low O(2) condition. PMID- 12902905 TI - Study of local three-dimensional conformal radiotherapy combined with transcatheter arterial chemoembolization for patients with stage III hepatocellular carcinoma. AB - The purpose of this study was to evaluate treatment-related toxicity, outcome, patterns of failure, and prognostic factors for patients with stage III unresectable hepatocellular carcinoma (HCC) treated with a combination of local 3 dimensional conformal radiotherapy (3D-CRT) and transcatheter arterial chemoembolization (TACE) under the support of G-CSF. From October 1997 to August 2001, 45 patients with stage III unresectable hepatocellular carcinoma underwent transcatheter arterial chemoembolization with local 3D-CRT. Twenty-seven patients were classified as having stage IIIA disease according to the American Joint Committee on Cancer (AJCC) staging system and 18 were classified as stage IIIB. The mean diameter of the treated hepatic tumor was 8.5 cm. Before 3D-CRT, 2 cycles of transcatheter arterial chemoembolization were prescribed. Forty-eight hours later, the G-CSF was prescribed for 5 days after the completion of every TACE. With the interval of 10 to 14 days after the second cycle of TACE, 3D-CRT was prescribed to all patients with a total dose of 50.4 Gy at 1.8 Gy per fraction 5 days per week. After the completion of 3D-CRT, the additional 2 cycles of TACE were given. All patients were monitored for treatment-related toxicity, outcome, patterns of failure, causes of death, and prognostic factors. Forty-two of 45 patients were treated smoothly with the primary schedule. In a median follow-up period of 27 months, 22 patients were alive and 23 were dead. Progressive disease occurred in 28 patients, including local recurrence alone (4 patients), distant metastases with local recurrence (8 patients), and distant metastases alone (16 patients). Nine patients developed radiation-induced liver disease (RILD). Three patients had treatment-related gastrointestinal bleeding. There were 2 treatment-related deaths, including 1 from RILD and 1 from gastrointestinal bleeding. Complete regression (CR) was observed in 6 patients, partial regression (PR) in 35 patients, and stable disease (SD) in 4 patients. The median overall survival duration from treatment was 23.5 months with a 1-year overall survival rate of 68.5%, a 2-year survival rate of 48.3%, and a 3-year survival rate of 22.6%. The median freedom from progressive disease survival duration from treatment was 25 months with 1-year, 2-year, and 3-year progression free survival rates of 76.2%, 56.8%, and 42.4%, respectively. The stage of HCC, regional lymph node status, portal vein thrombosis, pretreatment alpha fetoprotein level (AFP), and tumor size affected the treatment outcomes significantly. Therefore, for patients with stage III unresectable hepatocellular carcinoma, combined local 3D conformal radiotherapy with transcatheter arterial chemoembolization under the support of G-CSF is an effective treatment protocol. Further research is required to decrease distant metastases and to determine the safe irradiation dose-volume. PMID- 12902906 TI - Concomitant boost of stratified target area with gamma knife radiosurgery: a treatment planning study. AB - Conventional Gamma Knife Stereotactic Radiosurgery (GKSRS) has been focused on delivering a single peripheral dose to the gross target volume based on the anatomic information derived from the magnetic resonance or computed tomography (CT) studies. In this study, we developed a treatment planning approach that allows a boost dose to be delivered concomitantly to the desired subtarget area while maintaining the peripheral isodose coverage of the target volume. The subtarget area is defined as the high-risk or the tumor burden areas based on the functional imaging information such as the magnetic resonance spectroscopy (MRS) studies or the physician's clinical diagnosis. Treatment plan comparisons were carried out between the concomitant boost plans and the conventional treatment plans using dose volume histogram (DVH), tissue volume ratio (TVR), and the maximum dose to the peripheral dose ratio (MD/PD) analysis. Using the concomitant boost approach, more conformal and higher dose was delivered to the desired subtarget area while maintaining the peripheral isodose coverage of the gross target volume (GTV). Additionally, the dose to the normal brain tissue was found to be equivalent between the concomitant boost plans and the conventional plans. As a result, we conclude that concomitant boost of a stratified target area is feasible for GKSRS. PMID- 12902907 TI - Transiently increased bone density after irradiation and the radioprotectant drug amifostine in a rat model. AB - At therapeutic levels in pediatric patients, radiation causes damage to the growth plate and contributes to growth deformity and fractures. The purpose of this project was to examine the effects of x-ray irradiation on regional bone mineral density (BMD) and osteoclast histology of rat bone with and without radioprotectant amifostine (AMF) pretreatment. Seventy-two weanling rats had their right knee irradiated with single fraction 17.5 Gy, whereas the left leg was used as an internal control. Twelve animals were euthanized at each of 6 time periods (0.5-6 wk) after irradiation, half having received 100 mg/kg amifostine. BMD (g/cm3) was determined for both the right and left femurs using peripheral quantitative computed tomography (CT) (pQCT). Tibial sections were stained for osteoclasts/chondroclasts with tartrate-resistant acid phosphatase. Statistically significant increases in BMD within the radiation field were seen in the treatment groups' right irradiated legs over the control unirradiated left legs at all time points from 0.5 through 6 weeks. Anatomically, a peak in BMD occurs in the region immediately adjacent to the chondro-osseous junction at 2 weeks after irradiation and then moves proximally within the adjacent metaphysis after 3 weeks. Corresponding to these findings, histologically a 2-week nadir occurs after irradiation in osteoclasts/chondroclast numbers adjacent to the chondro osseous junction with a 71.9% decrease compared with controls (p <0.05). At 3 weeks, the numbers of osteoclasts/chondroclasts in this region have increased to 47.4% greater than the control legs (p <0.03) The animals receiving amifostine had BMD that was consistently closer to controls only adjacent to the chondro osseous junction at 0.5, 2, and 3 weeks and osteoclast/chondroclast numbers that were closer to controls only at 4 weeks. PMID- 12902908 TI - Role of radiotherapy in the treatment of neurosarcoidosis. AB - Neurosarcoidosis is usually managed with steroids, immunosuppressives, and other medications. Several small series suggest that radiotherapy might be useful in patients whose disease is refractory to conventional treatment. The purpose of this article is to report the outcome of 4 patients with neurosarcoidosis who were treated at the University of Florida. With long-term follow up, partial regression of disease was observed in 2 patients, stabilization of disease in 1 patient, and disease progression in 1 patient. Our experience and review of the related literature suggest the following conclusions: (1) radiotherapy is often effective in preventing the progression of local symptoms from neurosarcoidosis, but has limited application in reversing established neurologic deficits; (2) sarcoid meningitis is responsive to radiotherapy; and (3) radiation dose for the palliation of symptoms related to neurosarcoidosis is 20 to 25 Gy. PMID- 12902910 TI - Prognostic implications of t(10;11) translocations in childhood acute myelogenous leukemia: a report from the Children's Cancer Group. AB - PURPOSE: This was a retrospective analysis of outcome based on cytogenetics for a Children's Cancer Group phase 3 trial of acute myelogenous leukemia (AML) (CCG 2891). PATIENTS AND METHODS: A retrospective analysis of outcome for newly diagnosed children with AML and myelodysplastic syndrome (MDS) was performed using data collected from CCG-2891. The authors identified 11 patients whose blasts carried t(10;11) reciprocal translocations or other complex rearrangements involving 10p and 11q among 470 eligible patients entered with acceptable, centrally reviewed cytogenetics. A bone marrow specimen was used for each case of cytogenetic analysis in which 20 banded (either G-banded or Q-banded) metaphases were completed on each subject. All 11 patients had characteristic monocytoid morphology (M4 or M5) and tended to be young (0.1-7.9 years; median 0.9 years). RESULTS: All 11 patients entered remission, but remissions tended to be short; 9 patients relapsed within 12 months (median 4 months). The relapse rate of 82% was significantly higher for this group of patients compared with 46% for the group at large. The relapse rate for this group of patients having t(10;11) reciprocal translocations or other complex rearrangements involving 10p and 11q was also significantly higher compared with subjects with other 11q23 chromosomal abnormalities. The CNS relapse rate of 55% was higher for this group of patients compared with 3% for all other patients in the study. The CNS relapse rate was higher for the subjects who had t(10;11) reciprocal translocations or other complex rearrangements involving 10p and 11q compared with subjects with all other chromosome 11 abnormalities. Three children survived, two in second remissions (4.7 and 6.3 years after relapse) and one in first remission (7.0 years after diagnosis). Survival and event-free survival for the patients with t(10;11) reciprocal translocations or other complex rearrangements involving 10p and 11q was 27 +/- 27% and 9 +/- 17% at 6 years, respectively, and was not statistically different from all other patients with cytogenetics. Similarly, the survival and event-free survival for the patients with t(10;11) translocations and other rearrangements of chromosomes 10 and 11 was 27 +/- 27% and 9 +/- 17% at 6 years, respectively, and was not statistically different from the 11q23 group of subjects. CONCLUSIONS: Further research is needed to determine the various changes that are occurring at the molecular level for patients with t(10;11) translocations and other rearrangements of chromosomes 10 and 11 to gain insight into the mechanisms causing this clinical phenotype associated with a poor prognosis. PMID- 12902911 TI - Oral topotecan for refractory and relapsed neuroblastoma: a retrospective analysis. AB - PURPOSE: Among patients with multiply relapsed neuroblastoma refractory to conventional chemotherapy, oral topotecan has often been used for palliation. Although toxicity was generally thought to be mild, the efficacy of such an approach remains unproven. METHODS: The authors retrospectively analyzed patients with multiply relapsed or refractory neuroblastoma who were treated with oral topotecan for palliation. Each course was generally 1 mg/m2/d in two divided doses, for 21 consecutive days, repeated after a 1-week rest in patients without symptoms of progressive disease. Disease status was assessed by radiographic studies, urine catecholamine levels, and multiple bone marrow aspirations and biopsies. RESULTS: Twenty patients between the ages of 3 and 34 (median 13 years) received 1 (n = 7), 2 (n = 3), 3 (n = 4), 4 (n = 2), 6 (n = 2), and 12 courses (n = 2). Prior treatments included multiple cycles of high-dose alkylator-based chemotherapy (n = 20), high-dose intravenous topotecan (n = 8), myeloablative chemotherapy or radioimmunotherapy (n = 10), or experimental biologic agents (n = 16). Anti-neuroblastoma effects were seen in five patients lasting 6 to 12 months; two additional patients remained stable for 4 months. Thirteen patients had progressive disease (11 after one or two cycles). Toxicity included diarrhea (n = 12) requiring a dose adjustment in three patients and discontinuation of the drug in a fourth, and myelosuppression (n = 11) requiring transfusion and/or granulocyte-colony stimulating factor support. CONCLUSIONS: Oral topotecan therapy has antitumor activity in a small percentage of patients with relapsed or refractory neuroblastoma. Toxicities, including diarrhea and myelosuppression, may necessitate a dose adjustment in this patient population. Low-dose oral topotecan may have utility in the treatment of neuroblastoma. PMID- 12902912 TI - Evaluation and treatment of fever in the non-neutropenic child with cancer. AB - PURPOSE: To document the current approach to the evaluation and treatment of febrile non-neutropenic children with cancer by surveying American Society of Pediatric Hematology/Oncology (ASPHO) members. METHODS: Five hundred ninety-five ASPHO members were sent questionnaires polling their current practices in the evaluation and treatment of clinically stable, febrile (oral temperature of 38 degrees C x2, or >or=38.3 degrees C), non-neutropenic (absolute neutrophil count > 500/microL) pediatric cancer patients who have no identifiable bacterial infection. Patients were assigned to one of four groups based on the presence or absence of a central venous catheter (CVC) and no source (NS) or presumed viral source (VS) for fever. RESULTS: Three hundred sixteen ASPHO members (53%) responded and 300 questionnaires were evaluable. If a CVC was present, respondents universally drew blood cultures (99% NS, 96% VS) from the CVC, but only one third drew peripheral blood cultures (33% NS, 29% VS). If a CVC was not present, the majority of respondents (80% NS, 61% VS) drew peripheral cultures. ASPHO members started empiric antibiotics more frequently in patients with a CVC (69% NS, 55% VS) than in patients without a CVC (33% NS, 23% VS). Most did not admit these patients to the hospital (40% CVC and NS, 33% CVC and VS, 22% no CVC and NS, 19% no CVC and VS). ASPHO members with more years of experience were more likely to obtain peripheral blood cultures if a CVC was present and to admit a patient without a CVC. CONCLUSIONS: Nearly all respondents obtained a blood culture from the CVC in the initial assessment of the febrile, non-neutropenic patient with a CVC, but other evaluation and management practices varied considerably. PMID- 12902913 TI - Immediate versus delayed access of implantable venous access devices: does the timing of access make a difference to the frequency of complications? AB - PURPOSE: In many institutions protocols have not been developed as to when implantable venous access devices are accessed in children with cancer. The differences in complication rates (infection, hematoma, mechanical failure, and extravasation) between immediate versus delayed access remain unknown. PATIENTS AND METHODS: This retrospective study looks at the incidence of complications in two groups of pediatric patients who had an implantable venous access device inserted between 1998 and 2001 at McMaster Children's Hospital. Group 1 (immediate access group) had 23 patients and group 2 (delayed access) had 74 patients. RESULTS: The incidence of infection was 22% in group 1 and 14% in group 2. The difference between these infection rates was not statistically significant. All infections occurred in patients with a diagnosis of acute lymphoblastic leukemia. Of the patients in this study with acute lymphoblastic leukemia, 33% in group 1 and 36% in group 2 developed infections. CONCLUSIONS: These results suggest that implantable venous access devices can be accessed at the time of device insertion to decrease painful needle punctures in children with cancer and to provide secure immediate central venous access. PMID- 12902914 TI - Ethanol-lock technique in the treatment of bloodstream infections in pediatric oncology patients with broviac catheter. AB - PURPOSE: To assess the ethanol-lock technique as a means of treating central venous line infections. Bloodstream infections in patients with tunneled central venous catheters can lead to removal of the lines. METHODS: Twenty-eight children and adolescents aged 2 to 18 years, with different types of cancer, had Broviac catheters and presented with positive blood culture and clinical signs of infection between January 2000 and December 2001. The ethanol-lock technique was performed 24 times in 18 patients in addition to empiric (initially) and specific (after antibiogram) intravenous antibiotic treatment. In another 15 cases, 13 children were treated with systemic antibiotics alone. RESULTS: Sixty-seven percent of the patients treated with ethanol locks had no infectious relapse of any kind within 4 weeks of treatment or during subsequent aplasia, compared with 47% treated with systemic antibiotics alone. In one boy the catheter infection could not be cleared with systemic antibiotics alone, but after one course of ethanol locks no more blood culture-positive infectious episodes were observed. No severe clinical side effects of ethanol flush were observed. Mild symptoms that occurred were tiredness, headaches, dizziness, nausea, and light-headedness. CONCLUSIONS: The ethanol-lock technique appears to be a safe, well tolerated, and effective way to treat central venous line infections, even in small children. A prospective randomized study should be designed to compare antibiotic-lock, ethanol-lock technique, and systemic antibiotics alone in the treatment of device associated bloodstream infection. PMID- 12902915 TI - Alpha Thalassemia is associated with decreased risk of abnormal transcranial Doppler ultrasonography in children with sickle cell anemia. AB - PURPOSE: Cerebrovascular complications of sickle cell disease (SCD) are common, but the risk factors remain unclear. The multicenter Stroke Prevention Trial in Sickle Cell Anemia (STOP) provided an opportunity to examine alpha thalassemia-2 as a modifying risk factor, using abnormal transcranial Doppler ultrasonography (TCD) as a surrogate marker for cerebrovascular disease. The authors hypothesized that children with abnormal TCD are less likely to have alpha thalassemia-2, and an increased hemoglobin level accounts for this protective effect. METHODS: A retrospective study was conducted of children with SCD who had both alpha gene and TCD data from STOP: 128 with TCD of at least 200 cm/s (abnormal TCD) and 172 with TCD less than 170 cm/s (normal TCD). RESULTS: Alpha thalassemia-2 was more frequent in the normal TCD group compared with the abnormal TCD group. The odds ratio for normal TCD and alpha thalassemia-2 was 4.1. Adjusting for either hemoglobin level or red cell size (mean corpuscular volume) reduced the odds ratio only slightly. Age, normal TCD, and alpha thalassemia-2 had significant statistical interaction, so that alpha thalassemia-2 was not related to TCD for age 10 years or older. CONCLUSIONS: The frequency of alpha thalassemia-2 was significantly higher in children with normal TCD. Speculation on mechanisms of effect includes improved erythrocyte deformability, reduced red cell adhesion, and reduced nitric oxide scavenging in alpha thalassemia-2. The association of alpha thalassemia-2 and normal TCD adds to the evidence on the protective effects of alpha thalassemia-2 in SCD and highlights the contribution of epistatic factors. PMID- 12902916 TI - Hydroxyurea and arginine therapy: impact on nitric oxide production in sickle cell disease. AB - PURPOSE: Recent data suggest that hydroxyurea (HU) increases the production of nitric oxide (NO), a potent vasodilator. NO is normally metabolized from l Arginine (Arg). However, in vitro and animal experiments suggest that HU is the NO donor itself. In contrast, a recent study indicates that nitric oxide synthase (NOS) may play a role. Since adults with sickle cell disease (SCD) are Arg deficient, Arg availability may limit the ability of HU to maximally impact NO production if an NOS mechanism is involved. The authors have previously shown that Arg supplementation alone induces a paradoxical decrease in NO metabolite (NO(x)) production. METHODS: The authors studied the effects of HU and Arg supplementation on NO(x) production. HU alone or HU + Arg was administered to patients with SCD at steady state, and sequential levels of Arg, serum NO(x) and exhaled NO were followed over 4 hours. RESULTS: After HU + Arg, all patients demonstrated a significant increase in serum NO(x) production within 2 hours. When the same patients were treated with HU alone (5.1 +/- 2 micromol/L), a mixed response occurred. NO(x) levels increased in four patients and decreased in one patient (-23.3 micromol/L). CONCLUSIONS: While Arg alone does not increase serum NO(x) production in SCD patients at steady state, it does when given together with HU. Hence, co-administration of Arg with HU may augment the NO(x) response in SCD and improve utilization of Arg in patients at steady state. PMID- 12902917 TI - Ovarian carcinoma in an adolescent with transgenerational exposure to diethylstilbestrol. AB - Small cell carcinoma of the ovary is rare in adolescence. The etiology of such tumors is unknown. Several reports suggest that transgenerational exposure to diethylstilbestrol (DES) may have deleterious effects on the genitourinary tract. The authors describe a 15-year-old girl with small cell carcinoma of the ovary whose maternal grandmother had been taking DES while she was pregnant with the patient's mother. This case, while anecdotal, suggests that a transgenerational history of DES exposure should be noted, and that granddaughters with persistent abdominal pain even during childhood may need evaluation for genital tract abnormalities. PMID- 12902918 TI - Cholelithiasis and choledocholithiasis after sequential cytarabine and asparaginase. AB - We report 2 children with acute leukemia who developed symptomatic cholelithiasis and choledocholithiasis shortly after receiving sequential high-dose cytarabine and asparaginase. Known risk factors for gallstone formation were not present in our patients. These children did not experience any unusual therapy-related hepatic toxicity after laparoscopic cholecystectomy. PMID- 12902919 TI - Fetal rhabdomyomatous nephroblastoma: report of 14 cases confirming chemotherapy resistance. AB - The authors retrospectively reviewed their experience with 14 cases of fetal rhabdomyomatous nephroblastoma (15.6% of all renal tumors seen in the 1984-1998 period), analyzing the diagnostic and treatment approach as well outcome. Mean age at presentation was 27 months; 10 (71%) patients were younger than 36 months (5 of these were less than 12 months) at diagnosis. Three patients had bilateral tumors. One case was extrarenal. Nine patients received preoperative chemotherapy after fine-needle aspiration biopsy. The response was null in five children and mild in three; in the remaining one the tumor increased in size. The only patient showing greater than 50% tumor shrinkage was the one treated with radiotherapy. Postoperative treatment according to staging followed the SIOP protocol. In the follow-up (mean 59.5 months) of 12 patients, 6 were in stages 1-2: 4 are alive and free of disease and 2 died (one due to intractable local recurrence and the other in complete clinical remission). All six patients presenting in advanced stages died (four due to disease progression and two due to complications while in complete clinical remission). Patients with fetal rhabdomyomatous nephroblastoma presenting with distant metastatic disease and high stage have unfavorable outcomes, confirming data from the literature indicating that chemotherapy is not as effective as in other variants of nephroblastoma. New protocols are needed to overcome this situation, for which surgery appears the only truly effective option. PMID- 12902920 TI - Marked regression of metastatic pilocytic astrocytoma during treatment with imatinib mesylate (STI-571, Gleevec): a case report and laboratory investigation. AB - Pilocytic astrocytomas are the most common childhood glioma. Most children with pilocytic astrocytomas survive many years with their tumor, but alternative treatment approaches are needed for those with refractory or metastatic disease. Signaling by the platelet-derived growth factor tyrosine kinase receptor pathways have been postulated to contribute to the development of gliomas. The authors treated a single patient with refractory, metastatic pilocytic astrocytoma with the tyrosine kinase inhibitor imatinib mesylate and observed marked, transient regression of tumor during treatment. Immunohistochemistry was used to assess expression of reported target genes of imatinib mesylate in this patient's tumor tissue and of the PDGFR in pilocytic astrocytomas from 19 other patients. Immunohistochemistry showed that the patient's tumor cells did not express any of the reported target molecules inhibited by imatinib mesylate. PDGFR expression was detected in tumor vasculative in the panel of 20 tumors, and not in the tumor cells. The authors suggest that the PDGFR-signaling pathway postulated to contribute to the development of gliomas in adults might not contribute to pilocytic astrocytomas in children, and that treatment with imatinib mesylate should be considered in patients with refractory pilocytic astrocytoma. PMID- 12902921 TI - Neuroblastoma with expression of erythropoietin resulting in erythrocytosis. AB - We report the detection of erythropoietin mRNA in the tumor from a patient with neuroblastoma and erythrocytosis. A 2-year-old girl with neuroblastoma presented with hemoglobin 21.3 g/dL, red blood cells 8.03 x 1012/L, and hematocrit 0.641. The serum erythropoietin level was 26.54 mU/mL. The hemoglobin, hematocrit, and serum erythropoietin level were within normal ranges 3 months after surgical excision of the tumor. RT-PCR analysis showed that erythropoietin mRNA expression in the tumor tissue was as high as that of normal renal tissue. These results conclusively demonstrated that the tumor was the site of the ectopic erythropoietin production. PMID- 12902922 TI - Embryonal rhabdomyosarcoma presenting as conjunctival tumor. AB - The authors report the case of a 10-year-old girl who presented with a focal conjunctival lesion of the right eye. Conjunctival biopsy was performed and revealed an embryonal rhabdomyosarcoma. She received chemotherapy alone according to the Malignant Mesenchymal Tumor 95 Study (stage II nonalveolar orbital tumors) and is still in complete remission 6 months after the completion of treatment. Rhabdomyosarcomas of the conjunctiva without orbital extension are rarely reported. The authors discuss the prognosis and differential diagnosis of these lesions. PMID- 12902923 TI - Iris and anterior chamber involvement in acute lymphoblastic leukemia. AB - The authors describe a 6-year-old child, with a history of acute lymphoblastic leukemia, who presented with leukemic infiltration of the anterior segment of one eye mimicking anterior uveitis. After tissue diagnosis, administration of radiation therapy to the affected eye resulted in dramatic improvement of the condition. Involvement of the iris and anterior chamber is a recognized but unusual manifestation of leukemia. In the absence of systemic relapse, local radiation therapy without chemotherapy is highly effective, although the prognosis in these patients may be guarded. PMID- 12902924 TI - Mediastinal T-cell lymphoma in a boy 7 years after treatment of supratentorial primitive neuroectodermal tumor. AB - Medulloblastoma/primitive neuroectodermal tumor (PNET) is one of the most common central nervous system tumors in children and requires aggressive multimodality therapy. The authors describe a patient who developed mediastinal T-cell lymphoma 7 years after treatment of supratentorial PNET. Despite a good tumor response, the patient died during induction therapy because of invasive pulmonary aspergillosis. The authors conclude from this case that patients treated for medulloblastoma/PNET should be followed carefully for years. In addition, patients with second malignancies should be considered as a high-risk population for opportunistic infections. PMID- 12902925 TI - Intracranial hemorrhage in immune thrombocytopenic purpura: a retrospective analysis. AB - PURPOSE: To ascertain characteristics of children with immune thrombocytopenic purpura (ITP) and intracranial hemorrhage (ICH). METHODS: The authors identified 75 published cases of ICH in children with ITP by review of the literature from 1954 to 1998. Data pertaining to the ICH was recorded for age, gender, time from diagnosis of ITP (to ICH), platelet count, head trauma or arteriovenous malformation, concomitant medications, associated infections, other bleeding manifestations, prior treatment, and outcome. RESULTS Sixty-two cases represented 6 months to 20 years of age; 65% of patients were female. The median time from the diagnosis of ITP to ICH was 32 days (range 0 days to 8 years). Fifty of 69 ICH cases (72%) occurred within 6 months of diagnosis, but only 7 (10%) occurred within 3 days of diagnosis. The platelet count was less than 10000/microL in 71.4% of the cases. Treatment prior to the ICH was primarily steroids but also included intravenous immune globulin (IVIG), splenectomy, and others (interferon, azathioprine, or vincristine). There was no difference in mortality of patients before (56%) or after (54%) 1980. CONCLUSIONS: A very low platelet count appears permissive but not sufficient for ICH to occur in children with ITP. ICH occurs more commonly in acute ITP but can occur years after diagnosis. A significant number of patients develop an ICH despite having already initiated steroid treatment of ITP. PMID- 12902926 TI - Infection-associated hemophagocytic syndrome due to Pseudomonas aeruginosa in preterm infants. AB - Infection-associated hemophagocytic syndrome due to bacteria is rare and its appearance in preterm babies is uncommon. The signs, symptoms, laboratory values, and clinical status of three preterm babies with infection-associated hemophagocytic syndrome due to Pseudomonas aeruginosa are described and differences between preterm and infant cases are discussed in this report. PMID- 12902927 TI - Systemic lupus erythematosus in children with sickle cell disease. AB - Coexistence of sickle cell disease (SCD) and systemic lupus erythematosus (SLE) has been reported in 11 patients. The authors describe five additional patients with SCD and symptoms initially attributable to SCD who were later found to have SLE. Patients were identified over a 10-year period (1991-2001) in a pediatric sickle cell population numbering approximately 350. All patients are African American. Age at diagnosis of SLE was 9 to 17 years (median 11 years), and follow up after diagnosis ranged from 6 months to 10 years (median 3 years). SLE cerebritis (n = 3), serositis (n = 4), and nephritis (n = 2) were common findings. Physicians should be alerted to the possible development of SLE in patients with SCD. PMID- 12902929 TI - ITP and empiric steroid treatment. PMID- 12902928 TI - Parotid carcinoma after autologous bone marrow transplantation for relapsed nephroblastoma. AB - Abdominal irradiation, especially if associated with doxorubicin administration, increases the risk of a secondary malignant neoplasm (SMN) after treatment of nephroblastoma. Secondary malignant salivary tumors are rare and usually occur in patients with previous cranial irradiation. The authors describe the case of a parotid mucoepidermoid carcinoma arising 13 years after diagnosis of nephroblastoma. This patient showed no characteristics reported in the literature as statistically significant for the development of an SMN. The authors believe that long-term, regular clinical examination is necessary even in patients at low risk of developing an SMN. PMID- 12902930 TI - Fatal Aeromonas hydrophila septicemia in a 16-year-old patient with thalassemia. PMID- 12902931 TI - Amino acid sequence homology of thrombopoietin and erythropoietin may explain thrombocytosis in children with iron deficiency anemia. PMID- 12902932 TI - Lung mass in a child. PMID- 12902933 TI - Help honor the profession. PMID- 12902934 TI - Guard against conflict of interest. PMID- 12902935 TI - Oncologists and end-of-life care. PMID- 12902936 TI - NPs speak out against AAP policy. PMID- 12902937 TI - Improve adolescents' access to emergency contraception. PMID- 12902938 TI - Chronic low back pain management in primary care. PMID- 12902939 TI - Better outcomes for depressed patients. PMID- 12902940 TI - Oxytrol approved for overactive bladder. PMID- 12902941 TI - Oxygen free radicals and the disease process. PMID- 12902942 TI - A new treatment for morbid obesity. PMID- 12902943 TI - Practice powder-free and latex safe. PMID- 12902944 TI - Development of an NP role in interventional radiology. PMID- 12902945 TI - Cervical arthroplasty in the management of spondylotic myelopathy. AB - Cervical spinal cord compression managed via an anterior approach with an arthrodesis is associated with a decreased range of motion and accelerated adjacent segment degeneration. Artificial cervical disc replacement may address these problems. A series of seven cases (three women, two men; age 31-55 years) of anterior cervical decompression and placement of an artificial disc prosthesis is presented. A total of nine Bryan cervical disc prostheses (Spinal Dynamics Corp., Mercer Island, WA) were placed. Clinical and radiologic follow-up was performed at 24 hours, 6 weeks, 3 months, 6 months, and then yearly (mean follow up period 6.29 months, range 1-17 months). There were no complications. There was an improvement in Nurick grade by 0.72 grade (P < 0.05) and Oswestry Neck Disability Index score by 51.4 points (P < 0.0001). Motion was preserved. Improvement in lordosis occurred in 29% of cases. The spinal cord was decompressed on postoperative imaging. Cervical arthroplasty after anterior cervical decompression at one or more levels represents an exciting tool in the management of spinal cord compression secondary to spondylotic disease or acute disc prolapse. This is the first study that looks specifically at cervical arthroplasty for cervical myelopathy. Longer follow-up will reveal any delayed problems with artificial disc implantation, but in the short term, this technique offers an excellent outcome. PMID- 12902946 TI - Biomechanical testing of an artificial cervical joint and an anterior cervical plate. AB - An in vitro biomechanical study was conducted to determine the effects of fusion and nonfusion anterior cervical instrumentation on cervical spine biomechanics in a multilevel human cadaveric model. Three spine conditions were studied: harvested, single-level artificial cervical joint, and single-level graft with anterior cervical plate. A programmable testing apparatus was used that replicated physiologic flexion/extension and lateral bending. Measurements included vertebral motion, applied load, and bending moments. Relative rotations at the superior, implanted, and inferior motion segment units (MSUs) were normalized with respect to the overall rotation of those three MSUs and compared using a one-way analysis of variance (P < 0.05). Application of an anterior cervical plate decreased motion across the fusion site relative to the harvested and artificial joint spine conditions. The reduced motion was compensated for by an increase in motion at the adjacent segments. Use of an artificial cervical joint did not alter the motion patterns at either the instrumented level or the adjacent segments compared with the harvested condition for all modes of testing. PMID- 12902947 TI - Partial disc replacement with the PDN prosthetic disc nucleus device: early clinical results. AB - The aim of this study was to evaluate the efficacy of the PDN prosthetic disc nucleus device for the treatment of chronic discogenic back pain caused by degenerative disc disease. Among the 48 patients who underwent nucleus replacement surgery from January 2001 through May 2002, 46 patients were followed >6 months. The mean Oswestry Disability Index score was 58.9% preoperatively, and it improved to 18% at the 1-year follow-up. Visual Analogue Pain Scale scores improved from a preoperative mean of 8.5 to 3.1 after 1 year. The mean Prolo Scale score also improved from 5.2 preoperatively to 7.2 at 1 year. Major complications included four cases of device migration, requiring revision surgery, and infection in one patient. According to MacNab's criteria, results were excellent in 5 patients (10.9%), good in 31 (67.4%), fair in 3 (6.5%), and poor in 7 (15.1%). The overall clinical success rate was 78.3%. Nucleus replacement with the PDN device seemed to be effective in treating patients with chronic discogenic back pain caused by degenerative disc disease. PMID- 12902948 TI - Prosthetic disc nucleus (PDN) replacement for lumbar disc herniation: preliminary report with six months' follow-up. AB - A clinical follow-up study was performed to evaluate lumbar disc nucleus replacement clinically and functionally. The objective was to assess the clinical efficacy of the prosthetic disc nucleus (PDN) for treatment of lumbar disc herniation. The PDN was designed to treat patients suffering from disc herniation and/or degeneration. Clinical trials for PDN were first conducted in 1996, and the results showed that the PDN device was effective in most of the patients who were implanted. These studies had been carried out with patients who were implanted with a pair of the devices. The main complication was device migration. Therefore, a follow-up study based on a single PDN device implantation was designed to evaluate its clinical result. Forty-five patients with lumbar disc herniation were implanted with one PDN device from March 2002 to December 2002. Thirty (66.7%) patients came to the clinical, functional, and radiographic 6 month follow-up examinations. Independent analysis was performed by careful review of the charts, operative notes, preoperative and postoperative radiographs, magnetic resonance images, and follow-up records of all patients. After implantation, significant proportions of patients experienced pain relief. Improvements were noted in pain intensity, walking distance, lumbar mobility, neurologic weakness, Oswestry and Prolo scores, and intervertebral disc height. No difference in work status after PDN implantation could be detected. Compared with the preoperative height, the intervertebral disc had gained 19.7% (P < 0.001). Device migration, failure, and dislocation were not noticed in any patient. A few patients had minor complications such as transient low-grade fever. Clinically, a single PDN is preferred and can effectively increase the height of the intervertebral disc in patients with lumbar disc herniation. However, long-term follow-up of PDN implantation needs to be studied. PMID- 12902949 TI - The fate of the adjacent motion segments after lumbar fusion. AB - Lumbar spine fusion is a commonly performed procedure in various pathologic conditions of the spine. Its role remains debated, and moreover, delayed complications may occur, among which is transitional segment alteration leading to recurrence of back pain, gross instability, and neurologic symptoms. Little is known about the long-term prevalence of this complication because of a lack of specific studies. We analyzed the fate of the transitional segments in a homogeneous group of patients operated on during a 14-year period for degenerative conditions of the lumbar spine resistant to conservative treatment. Follow-up ranged from 2 to 15 years. Seventy-five percent of our study group had a minimal 5-year follow-up. In this subgroup, 41% of the patients developed transitional segment alterations, and 20% needed a secondary operation for extension of the fusion. Potential risk factors such as postoperative delay, length of fusion, and spine imbalance were recognized. The frequency of delayed alterations of the adjacent segment and the severity of symptoms related to this complication in this study raise questions about the justification of fusion procedures in degenerative conditions of the spine without threatening instability. Data from the literature confirm the severity of the problem, but many uncertainties remain because of the lack of homogeneous and complete data on both the normal evolution of motion segments of the lumbar spine with age and the fate of the same segments when transformed in transitional segments. Future prospective studies on the subject are needed and must deal with homogeneous groups of patients. More reconstructive surgical procedures need to be developed to lessen the need for fusion procedures. PMID- 12902950 TI - Shock absorption in lumbar disc prosthesis: a preliminary mechanical study. AB - Lumbar disc prostheses have been used in treating symptomatic degenerative disc diseases. A few prostheses of the ball-socket design are currently available for clinical use, the joint mechanism being materialized either with a hard polymer core or a metal-to-metal couple. Other prostheses of "shock absorber" design were not available at the time of the study. The objective of this work was to establish whether there was a difference in the shock absorption capacity between a device having an ultra-high-molecular-weight polyethylene center core and a device having a metal-on-metal bearing. Vibration and shock loading were applied to two lumbar total disc prostheses: PRODISC, manufactured by Spine Solutions, and MAVERICK Total Disc Replacement, manufactured by Medtronic Sofamor Danek. The shock absorption capacity of the device was evaluated by comparing the input and the output force measurements. The disc prosthesis was mounted onto a test apparatus. Each side of the device was equipped with a force sensor. The input shock load and the output resulting forces were simultaneously measured and recorded. The loading force pattern included 1). a static preload of 350 N plus an oscillating vibration of 100 N with frequency sweeping from 0 to 100 Hz and 2). a sudden shock load of 250 N applied over a 0.1-second interval. Both input and output signal data were processed and were transformed into their frequency spectrums. The vibration and shock transmissibility of the device, defined as the ratio of the output spectrum over the input spectrum, were calculated in sweeping the frequency from 0 to 100 Hz. The phase deviation was calculated to characterize the shock absorber effects. For both tested devices under vibration and shock loading, the phase angle displacement between the input and the output signals was 10 degrees. Under oscillating vibration loading, both tested devices had a transmission ratio higher than 99.8%. Over the frequency interval 1-100 Hz, the difference in transmission ratio between the two devices was <0.3%. Under sudden shock loading, both tested devices had a transmission ratio higher than 98%. The difference between the two devices was <0.8%. Both tested devices have identical vibration and shock transmissibility. PMID- 12902951 TI - Lumbar spine arthroplasty: early results using the ProDisc II: a prospective randomized trial of arthroplasty versus fusion. AB - This study represents the first 39 patients with at least 6-month follow-up enrolled in a prospective randomized Food and Drug Administration study evaluating the safety and efficacy of the ProDisc II versus the control, a 360 degrees lumbar spinal fusion. Data were collected preoperatively and at 6 weeks, 3 months, and 6 months postoperatively. Visual Analog Scale (VAS), Oswestry Low Back Pain Disability Questionnaire (ODQ), and patient satisfaction rates were evaluated at these intervals, as well as range of motion, return to work, and recreational and ambulatory status. There were 28 ProDisc patients and 11 who underwent fusion. Six patients had two-level surgery. Estimated blood loss (ProDisc = 69 mL versus fusion = 175 mL) and operative time (ProDisc = 75 minutes versus fusion = 219 minutes) were significantly different (P < 0.01). Hospital stays were shorter (ProDisc = 2.1 days versus fusion = 3.5 days [P < 0.01]) for ProDisc patients. There was a significantly greater reduction in the ODQ scores at 3 months in the ProDisc group compared with the fusion group (P < 0.05). No difference was noted in VAS. A trend was identified at 6 months in patient satisfaction rates favoring ProDisc versus fusion (P = 0.08), and motion was significantly improved in ProDisc patients compared with the fusion group (P = 0.02). Ambulatory status as well as recreational activity improved faster in the ProDisc group. The data suggest that total disc arthroplasty may be an attractive option as opposed to lumbar fusion for the surgical treatment of disabling mechanical low back pain secondary to lumbar disc disease. PMID- 12902952 TI - Lumbar disc replacement: preliminary results with ProDisc II after a minimum follow-up period of 1 year. AB - Total disc replacement has the potential to replace fusion as the gold standard surgical treatment of degenerative disc disease. Potential advantages of disc replacement over fusion include avoidance of pseudarthrosis, postoperative orthoses, and junctional degeneration. After observing satisfactory clinical results at 7-11 years' follow-up with the ProDisc first-generation implant, a second-generation prosthesis was designed. This study is a prospective analysis of the early results of total disc replacement with the ProDisc II total disc prosthesis. Fifty-three patients had single-level or multilevel disc replacement and were evaluated clinically and radiographically preoperatively and at mean 1.4 year follow-up. There were clinically and statistically significant improvements in back and leg pain Visual Analog Scale and Oswestry disability scores that were maintained at final follow-up. The clinical results of patients with single- and multilevel surgery were equivalent. Satisfactory results were achieved in 90% of patients who had previous lumbar surgery. Complications occurred in 9% of patients and included vertebral body fracture, transient radicular pain, implant malposition, and transient retrograde ejaculation. Three patients (6%) required reoperation to address complications. No mechanical failure of the implants or loosening was observed, and the prostheses retained motion. Randomized, prospective, long-term studies will be necessary to compare the effectiveness of arthrodesis with total disc replacement. PMID- 12902953 TI - Complications of artificial disc replacement: a report of 27 patients with the SB Charite disc. AB - Disc prosthesis surgery is rapidly becoming an option in treating patients with symptomatic degenerative disc disease. Only short-term and midterm results are described in the literature. Most operated patients belong to the age group of 30 50 years. In these active patients, complications can be expected to increase with longer follow-up, similar to total joint replacements in the extremities. Reported here is a series of 27 patients from another institution, who presented with unsatisfactory results or complications after SB Charite disc replacement. The objective of this work was to describe the possible short- and long-term unsatisfactory results of disc prosthesis surgery. Twenty-seven patients were seen in a tertiary university referral center with persisting back and leg complaints after having received a Charite disc prosthesis. All patients were operated on in a neighboring hospital. Most patients were operated on at the L4 L5 and /or the L5-S1 vertebral levels. The patients were evaluated with plain radiography, some with flexion-extension x-rays, and most of them with computed tomography scans. The group consisted of 15 women and 12 men. Their mean age was 40 years (range 30-67 years) at the time of operation. The patients presented to us a mean of 53 months (range 11-127 months) following disc replacement surgery. In two patients, an early removal of a prosthesis was required and in two patients a late removal. In 11 patients, a second spinal reconstructive salvage procedure was performed. Mean follow-up for 26 patients with mid- and long-term evaluation was 91 months (range 15-157 months). Early complications were the following: In one patient, an anterior luxation of the prosthesis after 1 week necessitated removal and cage insertion, which failed to unite. In another patient with prostheses at L4-L5 and L5-S1, the prosthesis at L5-S1 dislocated anteriorly after 3 months and was removed after 12 months. Abdominal wall hematoma occurred in four cases. Retrograde ejaculation with loss of libido was seen in one case and erection weakness in another case. A temporary benefit was experienced by 12 patients, while 14 patients reported no benefit at all. Main causes of persistent complaints were degeneration at another level in 14, subsidence of the prosthesis in 16, and facet joint arthrosis in 11. A combination of pathologies was often present. Slow anterior migration was present in two cases, with compression on the iliac vessels in one case. Polyethylene wear was obvious in one patient 12 years after operation. In eight cases, posterior fusion with pedicle screws was required. In two cases, the prosthesis was removed and the segment was circumferentially fused. These procedures resulted in suboptimal long-term results. In this relatively small group of patients operated on with a Charite disc prosthesis, most problems arose from degeneration of other lumbar discs, facet joint arthrosis at the same or other levels, and subsidence of the prosthesis. It is to be expected that many more patients will be seen with late problems some years after this operation as the survivorship will decrease with time. PMID- 12902954 TI - Classification of heterotopic ossification (HO) in artificial disk replacement. AB - There is currently no structured classification system to quantitate heterotopic bone formation after artificial disk replacement procedures. The purpose of this work was to develop a method of classifying heterotopic bone formation that is reliable between investigators with different levels of training and easy to remember with only five gradations of severity. One hundred one radiographs of clinical patients and 17 microradiographs from nonhuman primates having undergone various types of disk replacement were classified by seven independent reviewers. The kappa statistics were calculated for interobserver variation between the seven participants with various levels of spinal training and the intraobserver error based on two assessments made at least 2 months apart. The interobserver reliability correlation coefficient for seven raters calculated using the intraclass kappa correlation coefficient and the Kish rho was r = 0.9683 (P < 0.0001). The intraobserver reliability based on readings at two time intervals at a minimum of 2 months apart was r = 0.8949 (P = 0.01). This classification of heterotopic ossification, periannular calcification, and ectopic bone formation associated with total disk arthroplasty proved to be highly reliable and reproducible. PMID- 12902955 TI - Treatment of juxtafusional degeneration with artificial disc replacement (ADR): preliminary results of an ongoing prospective study. AB - Eleven patients with juxtafusional degeneration syndrome underwent artificial disc replacement (ADR). Of these, five patients followed up for >6 months were analyzed for the current report. Surgery for juxtafusional degeneration was performed mean 4.9 years from their initial operation. The presenting features were neurologic compromise in four and severe back pain in one, with a mean Oswestry Disability Index (ODI) score of 64%. The replaced levels were L1-L2 in one, L3-L4 in three, L4-L5 in one, and L5-S1 in one. L1-L2 replacement was performed through a left flank approach. At 6 months, all patients had complete resolution of neurologic symptoms, and ODI decreased to 24%. ADR shows promising early results in juxtafusional degeneration in well-selected patients. PMID- 12902956 TI - The Anterolateral TransPsoatic Approach (ALPA): a new technique for implanting prosthetic disc-nucleus devices. AB - Prosthetic disc-nucleus devices have traditionally been implanted via the same posterior approach that is standard for lumbar decompression surgery. To avoid disrupting the posterior structures of the spine and to facilitate device implantation, the AnteroLateral transPsoatic Approach (ALPA) has been developed. With ALPA, an incision is made in the lateral region overlying the disc, and the lateroabdominal muscles are bluntly dissected. The retroperitoneal area is accessed and followed to the psoas muscle, where blunt dissection in the direction of the muscle fibers is used to reach the affected disc. The disc anulus is cut to create a flap, the nucleus pulposus material is removed, and the prosthesis is implanted into the disc cavity. The procedure concludes with the anulus flap being sutured in place over the opening to the nucleus, and the wound is closed in a normal manner. A small study group of eight patients has been implanted with PDN prosthetic disc-nucleus devices by way of the ALPA technique. Preliminary results indicate minor adverse events related to the surgical technique. Clinical results are also encouraging with significant improvements in Oswestry and Prolo scores and with an associated increase in disc height. The ALPA technique is a good alternative to the posterior hemilaminotomy approach for implanting nucleus devices in degenerated lumbar discs. PMID- 12902957 TI - A memory coiling spiral as nucleus pulposus prosthesis: concept, specifications, bench testing, and first clinical results. AB - The intradiscal cavity left after a discectomy can be filled by a new nucleus prosthesis made of polycarbonate urethane in the form of a "memory coiling spiral." Biomechanical tests have demonstrated that this device compensates for the loss of disc height, decreases the compression of the facet joints, and restores the kinematics of the spinal segment, without deformation of the vertebral endplates or migration. The device is currently under clinical investigation. Inclusion and exclusion criteria of the pilot study are presented, and preliminary results of the first five patients supplied with the spiral are reported after an average follow-up time of 24 months. No migration of the device has been observed so far. With its easy application due to the standardized approach and the memory coiling mechanism, this device represents an advance within the nonfusion techniques. PMID- 12902958 TI - The implications of constraint in lumbar total disc replacement. AB - Lumbar total disc replacement (TDR) is an evolving technique that has the potential to replace arthrodesis as the gold standard surgical treatment of degenerative disc disease. The interaction between host anatomy and physiology and the biomechanical properties of TDR implants will determine the quality of long-term clinical results. However, there is scant literature addressing this subject. The purpose of this article is to discuss the implications of biomechanical constraint in TDR. Based upon available data for normal motion segments and the design of two TDRs currently in clinical trials, unconstrained designs appear to have a kinematic advantage. They are more likely to provide a physiologic mobile instantaneous axis of rotation (IAR), which may explain why they display greater range of motion in vivo. Their lack of constraint may prevent excessive facet joint or capsuloligamentous loads in the extremes of flexion and extension. Furthermore, since the IAR is mobile, they may be less sensitive to small errors in implant placement. On the other hand, constrained devices appear to have an advantage in protection of the posterior elements from shear loading. Spinal shear loads of considerable magnitude occur during activities of daily living. Whether the transference of stresses to the implant and implant-bone interface is clinically significant is unknown. Although this article focuses on two specific TDR designs, future designs will need to account for the same kinematic and loading concerns regarding constraint. We hope this discussion will assist clinicians and researchers in the design, selection, and clinical comparison of present and future TDR implants. PMID- 12902959 TI - Dynamic stabilization of the lumbar spine and its effects on adjacent segments: an in vitro experiment. AB - In recent years, nonfusion stabilization of the lumbar spine has gained more and more popularity. These nonfusion systems intend to maintain or restore the intersegmental motions to magnitudes of the intact spine and have no negative effects on the segments adjacent to the stabilized one. This study investigated the DYNESYS, a dynamic nonfusion system, which is designed to stabilize the bridged segments while maintaining the disc and the facet joints. To determine the magnitude of stabilization and the effect of the stabilization on the adjacent segment, six lumbar cadaver spines were fixed in a spine tester and loaded with pure moments in the three main motion planes. For each spine, four different stages were tested: intact, defect of the middle segment, fixation with the DYNESYS, and fixation with the internal fixator. Intersegmental motions were measured at all levels. For the bridged segment, the DYNESYS stabilized the spine and was more flexible than the internal fixator. This difference between the internal fixator and the DYNESYS was most pronounced in extension (P < 0.05), with the DYNESYS restoring the motion back to the level of the intact spine. The motion in the adjacent segments was not influenced by either stabilization method. Our results suggest that the DYNESYS provides substantial stability in case of degenerative spinal pathologies and can therefore be considered as an alternative method to fusion surgery in these indications while the motion segment is preserved. PMID- 12902960 TI - SB Charite disc replacement: report of 60 prospective randomized cases in a US center. AB - Sixty patients with one-level discogenic pain confirmed by plain radiography, magnetic resonance imaging, and provocative discography for degenerative disc disease were randomized: one-third BAK anterior interbody fusion and two-thirds anterior SB Charite artificial disc replacement. The mean age was 40.3 years (range 21-56 years). Nineteen cases were at L4-L5 and 41 cases were at L5-S1. Nineteen cases had BAK anterior interbody fusion and 41 cases were randomized as SB Charite disc replacement. The length of surgery was mean 88.4 minutes (range 54-137 minutes) for both groups. The estimated blood loss was mean 289.5 mL (range 50-1800 mL). The length of hospital stay was a mean of 3.03 days (range 2 6 days). Oswestry Disability Index scores for the SB Charite disc (aggregate study group) were 50.0 +/- 14.3 preoperatively and 25.0 +/- 20.1 at 1-3 years' follow-up (P < 0.001). This is the first study that shows improvement of functional outcome measures in a prospective randomized design with disc arthroplasty treating primarily mechanical back pain and achieving comparable successful results to lumbar fusion-interbody fusion cage and BMP or interbody autograft and pedicle screw instrumentation. PMID- 12902961 TI - Cervical spine geometry correlated to cervical degenerative disease in a symptomatic group. AB - OBJECTIVE: To investigate whether a statistical correlation exists between lateral cervical geometry and cervical pathology, as identified on neutral anteroposterior (AP) and lateral radiographs within a symptomatic group; describe the cervical pathology and determine its location and frequency; and identify the subject's age, sex, and chief complaint. SETTING: Department of radiology at a chiropractic college. METHODS: One hundred eighty-six consecutive pairs of AP and lateral cervical radiographs were reviewed for pathology. A 5-category severity scale was used to describe degenerative joint disease, the most common pathological finding. The subject's age, sex, and symptoms were recorded. Geometric analysis was focused on vertebral position, alignment, and gravitational loading acquired from the neutral lateral cervical radiograph. RESULTS: Regression and discriminant analysis identified 5 geometric variables that correctly classified pathology subjects from nonpathology subjects 79% of the time. Those variables were: (1) forward flexion angle of the lower cervical curve; (2) gravitational loading on the C5 superior vertebral end plate; (3) horizontal angle of C2 measured from its inferior vertebral end plate; (4) disk angle of C3; and (5) posterior disk height of C5. Degenerative joint disease was the most common pathological finding identified within discrete age, sex, and symptom groups. CONCLUSION: We identified 5 geometric variables from the lateral cervical spine that were predictive 79% of the time for cervical degenerative joint disease. There were discrete age, sex, and symptom groups, which demonstrated an increased incidence of degenerative joint disease. PMID- 12902962 TI - Changes in vertebral artery blood flow following normal rotation of the cervical spine. AB - BACKGROUND: Extreme rotation of the cervical spine may cause compromised vertebral artery (VA) blood flow. This is of particular interest to manual therapists because of the potential risks associated with these movements. The question is whether the decreased blood flow is significant and therefore likely to cause vertebrobasilar insufficiency/ischemia (VBI) and whether contralateral and ipsilateral rotations are equally affected. Several studies measuring VA blood flow have been reported. However, different parts of the VA were studied, in small samples of normal subjects and patients over a wide range of ages. Hence, the results are controversial. OBJECTIVE: To investigate intracranial VA blood flow in normal male subjects and female subjects, aged 20 to 30 years, in neutral and maximally rotated cervical spinal positions. METHODS: Transcranial Doppler sonography was used to measure intracranial VA blood flow, with the cervical spine in neutral and then rotated maximally to the left and later to the right. The sample consisted of 60 male subjects and 60 female subjects (240 VAs). Comparisons between the neutral and rotated head positions were made within and between the groups. RESULTS: There was a significant decrease (P =.001) in intracranial VA blood flow following cervical spine rotation, irrespective of side but greater on the contralateral side, in the total sample and in male subjects. Female subjects had a significantly higher blood flow than male subjects, and although they showed a significant decrease with contralateral rotation, there was no significant difference in blood flow on the ipsilateral side. CONCLUSION: Maximal rotation of the cervical spine may significantly affect vertebral artery blood flow, particularly when used in the treatment of patients with underlying vascular pathology. PMID- 12902963 TI - Lateral cervical curve changes in patients receiving chiropractic care after a motor vehicle collision: a retrospective case series. AB - OBJECTIVE: To examine radiological changes of the lateral cervical curve in patients who received chiropractic care after motor vehicle collisions. DESIGN: A retrospective case series. Thirteen patients who had received chiropractic care after motor vehicle collisions were selected from a northeastern Washington chiropractic office. Patients had a lateral cervical radiograph taken prior to the initiation of chiropractic treatment and a comparative lateral cervical radiograph subsequent to a period of care. Cases were included if they met the previously stated criteria and if the radiographs were of sufficient quality to determine the lateral cervical curve from C2-C7. RESULTS: Adjustments rendered using an Activator Adjusting Instrument. Eleven of the subjects were also instructed to perform stretching exercises. Compared to the initial lateral cervical radiograph, the comparative radiographs demonstrated a mean increase in cervical lordosis between C2 and C7 of 6.4 degrees (SD = 8.2). The standard error estimate of the population was 2.3 degrees, with a 95% confidence interval of 1.4 degrees to 11.4 degrees. CONCLUSION: There was a mean increase in the cervical lordosis of 6.4 degrees (SD = 8.2). The standard error estimate of the population was 2.3 degrees, with a 95% confidence interval of 1.4 degrees to 11.4 degrees. We were not able to determine the individual effects of adjustment, stretching, and natural progression of the condition. The results suggest that further study of this phenomenon should be undertaken. PMID- 12902964 TI - An investigation into the production of intra-articular gas bubbles and increase in joint space in the zygapophyseal joints of the cervical spine in asymptomatic subjects after spinal manipulation. AB - OBJECTIVES: To analyze the size and density of cervical zygapophyseal joint spaces in asymptomatic subjects using computed tomography (CT) and plain film radiography. The joint spaces were analyzed before manipulation without traction, before manipulation with manual traction, after manipulation without traction, and after manipulation with manual traction. The data obtained before the manipulation were compared with data obtained after the manipulation to determine if significant alterations occurred. DESIGN: Twenty-two asymptomatic subjects were placed into 1 or more of 6 possible experimental groups. In all experimental groups, except for experiment 1, the subjects underwent: (step 1) a premanipulation radiograph and/or CT scan, (step 2) a premanipulation plus traction radiograph and/or CT scan, (step 3) a postmanipulation radiograph and/or CT scan, and (step 4) a postmanipulation plus traction radiograph and/or CT scan. RESULTS: Except for experiment 2, there was no significant change in the width, area, and density values of the zygapophyseal joint spaces immediately after the manipulation in either the traction or traction-free positions of the neck, and no visible radiolucent cavities were demonstrated in any view. CONCLUSION: No evidence of gas in the joint space or obvious increase in zygapophyseal joint space width immediately after the manipulation was found. This was evident in both the traction and traction-free postmanipulation scans and plain film images. This is not consistent with the current understanding of cavitation in joints, in particular, the refractory period. PMID- 12902965 TI - Manual discrimination capability when only viscosity is varied in viscoelastic stiffness stimuli. AB - BACKGROUND: Training manual therapists in the judgment of spinal stiffness traditionally requires the spines of human volunteers. Presenting the range of stiffness values representative of spines requires both time and resources. When investigating the ability of therapists to judge spinal stiffness, several studies have therefore utilized spine analogues, nonbiological devices designed to present either purely elastic stimuli or viscoelastic stimuli with unknown relative contributions of elastic and viscous components. Previous research has suggested that the viscous component of stiffness is more difficult to judge. OBJECTIVE: This study determined for each subject the just-noticeable difference, or discrimination threshold, for manual judgment of viscous stiffness against a constant elastic and friction background. DESIGN: This psychophysical study utilized the Weber fraction as a measure of the ability of subjects to discriminate between viscoelastic stiffness stimuli. METHODS: Twenty-five subjects with both physiotherapy and lay backgrounds volunteered to participate in the study. Stiffness stimuli were generated by a device incorporating a fluid filled plunger and a spring, with only the amount of viscous stiffness being manipulated by the experimenter. The method of constant stimuli was used to estimate the just-discriminable change in viscous stiffness, and results were expressed as a percentage of the base stiffness or Weber fraction. RESULTS: The mean Weber fraction for manual judgments of viscous stiffness was 14.7%. For 13 of the subjects who had previously participated in elastic stiffness discrimination studies, the Weber fraction for viscous stimuli was significantly greater than that for elastic stimuli. CONCLUSION: The significantly higher Weber fraction for viscous stiffness perception compared with that for pure elastic stiffness suggests that the poor reliability of manual judgments of spinal stiffness may be due to the difficulty in judging the viscous stiffness component. PMID- 12902966 TI - Spinal palpation: The challenges of information retrieval using available databases. AB - PURPOSE: This study addressed 2 questions: first, what is the yield of PubMed MEDLINE for complementary and alternative medicine (CAM) studies compared to other databases; second, what is an effective search strategy to answer a sample research question on spinal palpation? METHODS: We formulated the following research question: "What is the reliability of spinal palpation procedures?" We identified specific Medical Subject Headings (MeSH) and key terms as used in osteopathic medicine, allopathic medicine, chiropractic, and physical therapy. Using PubMed, we formulated an initial search template and applied it to 12 additional selected databases. Subsequently, we applied the inclusion criteria and evaluated the yield in terms of precision and sensitivity in identifying relevant studies. RESULTS: The online search result of the 13 databases identified 1189 citations potentially addressing the research question. After excluding overlapping and nonpertinent citations and those not meeting the inclusion criteria, 49 citations remained. PubMed yielded 19, while MANTIS (Manual Alternative and Natural Therapy Index System), a manual therapy database, yielded 35 citations. Twenty-six of the 49 online citations were repeatedly indexed in 3 or more databases. Content experts and selective manual searches identified 11 additional studies. In all, we identified 60 studies that addressed the research question. The cost of the databases used for conducting this search ranged from free-of-charge to $43,000 per year for a single network subscription. CONCLUSIONS: Commonly used databases often do not provide accurate indexing or coverage of CAM publications. Subject-specific specialized databases are recommended. Access, cost, and ease of using specialized databases are limiting factors. PMID- 12902967 TI - Dietary diversity: case study of fruit and vegetable consumption by chiropractic patients. AB - BACKGROUND: A distinct health advantage has been linked with eating a wide variety of fresh fruit and vegetables. OBJECTIVE: This study explores the diversity of fruits and vegetables consumed by chiropractic patients and inquires into their preferred purchasing patterns. METHOD: A case study that included patients attending 24 chiropractic practices in 5 states and the Australian Capital Territory was undertaken. Seven hundred eighty-two patients were asked to complete a questionnaire providing general information about their life-style and particular information about the number of servings of fruits and vegetables that they usually consumed. RESULTS: Although overall chiropractic patients reported adhering to a number of healthy life-style behaviors, participants in this study would benefit from eating a greater diversity of fruits and vegetables. CONCLUSION: In view of the substantial health benefits associated with eating a wide variety of fruits and vegetables, it is suggested that chiropractic patients be encouraged to follow a color code system for purchasing their fresh produce. PMID- 12902968 TI - Chiropractic care of a pediatric patient with myasthenia gravis. AB - OBJECTIVE: To describe the chiropractic care of a pediatric patient with complaints associated with myasthenia gravis. Clinical features A 2-year-old girl was provided chiropractic care at the request and consent of her parents for complaints of ptosis and generalized muscle weakness (ie, lethargy), particularly in the lower extremities. Prior to entry into chiropractic management, magnetic resonance imaging of the brain and acetylcholine receptor antibody tests were performed with negative results. However, the Tensilon test was positive and the diagnosis of myasthenia gravis was made by a pediatrician and seconded by a medical neurologist. Intervention and outcome The patient was cared for with contact-specific, high-velocity, low-amplitude adjustments to sites of vertebral subluxation complexes in the upper cervical and sacral spine. The patient's response to care was positive and after 5 months of regular chiropractic treatment her symptoms abated completely. CONCLUSION: There are indications that patients suffering from disorders "beyond low back pain" as presented in this case report may derive benefits from chiropractic intervention/management. PMID- 12902969 TI - Developing skilled performance of lumbar spine manipulation. PMID- 12902970 TI - Developing skilled performance of lumbar spine manipulation. PMID- 12902973 TI - Vectored upper cervical manipulation for chronic sleep bruxism, headache, and cervical spine pain in a child. AB - OBJECTIVE: To discuss the management of chronic sleep bruxism in a 6-year old girl. Clinical features The patient had morning headaches and cervical spine pain. Due to abnormal tooth wear, bruxism had been previously diagnosed and was verified by observation during sleep. She also had abnormal postural and palpatory findings, indicating upper cervical joint dysfunction. Intervention and outcome Bilateral rotary cervical stretching/mobilization and a vectored high velocity, low-amplitude adjustment were performed in the upper cervical spine, using the atlas transverse process as the contact point. There was complete relief of the chronic subjective symptoms concomitant with remission of the objective signs of joint dysfunction. CONCLUSIONS: Cervical, particularly upper cervical, spine muscle-joint dysfunction should be considered as a potential etiology in chronic childhood sleep bruxism. PMID- 12902974 TI - Endocrine late effects of childhood cancer therapy. PMID- 12902976 TI - Modulation of human checkpoint kinase Chk1 by the regulatory beta-subunit of protein kinase CK2. AB - Protein kinase CK2 is a serine/threonine protein kinase involved in various aspects of cellular regulation. The regulatory beta-subunit of CK2 exerts a central role not only in mediating formation of tetrameric CK2 complexes but also as a docking partner for several protein kinases. In this study, CK2beta is found to interact with the human cell cycle checkpoint kinase Chk1. The Chk1 interacting region of CK2beta is localized at the C-terminus and the complex between CK2beta and Chk1 is devoid of the catalytic CK2alpha-subunit. The interaction between CK2beta and Chk1 leads to an increase in the Cdc25C phosphorylation activity of Chk1. The screening of several cell lines has revealed that the association between CK2beta and Chk1 also occurs in vivo at a different degree. Collectively, these studies confirm the implication of the regulatory beta-subunit of protein kinase CK2 in cell cycle regulation and identify a novel mechanism for the activation of Chk1 protein kinase. PMID- 12902977 TI - Inhibition of cell proliferation and induction of apoptosis by novel tetravalent peptides inhibiting DNA binding of E2F. AB - We have isolated several peptides from random peptide phage display libraries that specifically recognize the cell cycle regulatory transcription factor E2F and inhibit DNA binding of E2F/DP heterodimers (E2F-1, E2F-2, E2F-3, E2F-4 or E2F 5, and DP-1). The inhibitory efficiency could be strongly enhanced by generating branched tetravalent molecules. To analyse the biological consequences of peptide mediated E2F inhibition, we fused two of these branched molecules to a cell penetrating peptide derived from the HTV-Tat protein. Incubation of human tumor cells with these branched Tat-containing peptides led to an inhibition of cell proliferation and induction of apoptosis. These results provide new insights into the function of E2F and further validate E2F as a potential therapeutic target in proliferative diseases. PMID- 12902978 TI - The proteasome inhibitor PS-341 overcomes TRAIL resistance in Bax and caspase 9 negative or Bcl-xL overexpressing cells. AB - We demonstrate that PS-341, a small molecule inhibitor of the proteasome, markedly sensitizes resistant prostate, colon, and bladder cancer cells to TNF like apoptosis-inducing ligand (TRAIL)-induced apoptosis irrespective of Bcl-xL overexpression. PS-341 treatment by itself does not affect the levels of Bax, Bak, caspases 3 and 8, c-Flip or FADD, but elevates levels of TRAIL receptors DR4 and DR5. This increase in receptor protein levels is associated with the ubiquitination of the DR5 protein. When PS-341 is combined with TRAIL, the levels of activated caspase 8 and cleaved Bid are substantially increased. In Bax negative TRAIL-resistant HC-4 colon cancer cells, the combination of PS-341 and TRAIL overcomes the block to activation of the mitochondrial pathway and causes SMAC and cytochrome c release followed by apoptosis. Similarly, murine embryonic fibroblasts lacking Bax undergo apoptosis when exposed to the combination of PS 341 and TRAIL; however, fibroblasts lacking Bak are significantly resistant. Taken together, these findings indicate that PS-341 enhances TRAIL-induced apoptosis by increasing the cleavage of caspase 8, causing Bak-dependent release of mitochondrial proapoptotic proteins. PMID- 12902979 TI - Loss of inhibitor of apoptosis proteins as a determinant of polyamine analog induced apoptosis in human melanoma cells. AB - We have previously shown that the clinically relevant polyamine analog N1,N11 diethylnorspermine (DENSPM) causes rapid apoptosis in human melanoma SK-MEL-28 cells via a series of events that include mitochondrial release of cytochrome c and activation of the caspase cascade. Upstream to these events, DENSPM downregulates polyamine biosynthesis and potently upregulates polyamine catabolism at the level of spermidine/spermine N1-acetyltransferase (SSAT). In searching for downstream effectors that either contribute to or abrogate the apoptotic response, we observed that DENSPM treatment of SK-MEL-28 cells for 30 h led to cytosolic release of Smac/Diablo, a mitochondrial protein known to bind and inhibit the function of inhibitor of apoptosis proteins (IAPs). Subsequently, we found that DENSPM markedly lowered survivin and ML-IAP protein (but not XIAP) levels by 18 h via an apparently Smac/Diablo-independent pathway. Proteasome inhibitors fully prevented survivin and ML-IAP protein loss as well as apoptosis, suggesting that the proteasome-mediated degradation of survivin and ML-IAP is causally linked to the cellular outcome. We also observed that structural analogs of DENSPM which differentially induced SSAT and apoptosis lowered survivin and ML IAP levels in a manner that correlated with enzyme activity. The linkage between IAPs and SSAT was more directly established by the finding that selective prevention of SSAT induction by small interfering RNA prevented survivin and ML IAP loss as well as apoptosis during DENSPM treatment. Among the melanoma cell lines (SK-MEL-28, MALME-3M, A375 and LOX), survivin degradation correlated temporally with the onset of DENSPM induced apoptosis or growth inhibition. By contrast, ML-IAP degradation occurred only during rapid apoptosis seen in SK-MEL 28 cells. These data suggest a sequence of events whereby DENSPM induction of SSAT leads to loss of IAP proteins and a more fulminate apoptotic response. The findings implicate survivin and ML-IAP as important determinants of polyamine analog drug action in melanoma cells. PMID- 12902980 TI - The nuclear BAG-1 isoform, BAG-1L, enhances oestrogen-dependent transcription. AB - BAG-1 is a multifunctional protein that interacts with a wide range of cellular targets including heat-shock proteins and some nuclear hormone receptors. BAG-1 exists as three major isoforms, BAG-1L, BAG-1M and BAG-1S. BAG-1L contains a nuclear localization signal, which is not present in the other isoforms, and is predominantly localized in the cell nucleus. Here we have investigated the effects of BAG-1 on function of the oestrogen receptor (ER), a key growth control molecule and target for hormonal therapy in breast cancer. We demonstrate that BAG-1L, but not BAG-1S or BAG-1M, increased oestrogen-dependent transcription in breast cancer cells. BAG-1L interacted with and stimulated the activity of both ER alpha and beta. Although BAG-1L and ERs colocalize to the nucleus, fusing BAG 1S to an heterologous nuclear localization sequence was not sufficient to stimulate transcription. Consistent with an important effect on receptor function, nuclear BAG-1 expression in breast cancers was associated with expression of the progesterone receptor, a transcriptional target of ERalpha, and was associated with improved survival in patients treated with hormonal therapy. These data suggest that BAG-1L is an important determinant of ER function in vitro and in human breast cancer. PMID- 12902981 TI - Repression of MUC2 gene expression by butyrate, a physiological regulator of intestinal cell maturation. AB - Sodium butyrate (NaB) inhibits proliferation, stimulates apoptosis, and promotes differentiation of human colon cancer cells along the absorptive phenotype. In vitro, butyrate induces a switch from cells with a secretory to an absorptive phenotype. Here, we report that NaB specifically represses the expression of the MUC2 gene, a differentiation marker of the secretory goblet cell lineage, in forskolin- and 12-O-tetradecanoylphorbol 13-acetate-induced HT29 cells, and Cl.16E cells, a clonal derivative of HT29 cells that spontaneously differentiates into goblet cells. Thus, NaB repression is independent of the nature of the stimulus that triggers MUC2 expression. Further, repression was independent of new protein synthesis. Our results suggest that inhibition of MUC2 is linked to the ability of butyrate to repress histone deacetylase activity, since trichostatin A, another inhibitor of histone deacetylases, also inhibited MUC2 expression in induced HT29 cells. Finally, we demonstrate that the NaB effect is specific for this marker of the secretory cell lineage, since carcinoembryonic antigen, which is expressed in both the secretory and absorptive cells, is induced by NaB. Thus, the NaB repression of a definitive function of the secretory cell lineage is a further mechanism, in addition to the effects on proliferation and apoptotic pathways, through which butyrate can regulate intestinal homeostasis. PMID- 12902982 TI - Myc and E2F1 induce p53 through p14ARF-independent mechanisms in human fibroblasts. AB - p19ARF is induced in response to oncogene activation or during cellular senescence in mouse embryo fibroblasts, triggering p53-dependent and p53 independent cell cycle arrest and apoptosis. We have studied the involvement of human p14ARF as a regulator of p53 activity in normal human skin fibroblasts (NHFs) or WI38 lung embryonic fibroblasts expressing conditional Myc or E2F1 estrogen receptor fusion proteins. Both Myc and E2F1 activation rapidly induced p53 phosphorylation at Ser-15, p53 protein accumulation, and upregulation of the p53 target genes MDM2 and p21. Activation of E2F1 induced p14ARF mRNA and protein levels. In contrast, Myc activation did not induce any significant increase in p14ARF mRNA or protein levels in neither NHFs nor WI38 fibroblasts within 48 h. Myc and E2F1 induced p53 and cell cycle arrest even after silencing of p14ARF using short-interfering RNA. Treatment with the ATM/ATR kinase inhibitor caffeine prevented p53 accumulation upon activation of Myc or E2F1. Our results indicate that p53 phosphorylation, but not p14ARF, plays a major role for the induction of p53 in response to Myc and E2F1 activation in normal human fibroblasts. PMID- 12902983 TI - Use of RNA interference to validate Brk as a novel therapeutic target in breast cancer: Brk promotes breast carcinoma cell proliferation. AB - Brk (PTK6) is a nonreceptor protein tyrosine kinase, which is expressed in over 60% of breast carcinoma tissue samples and breast tumour cell lines, but not normal mammary tissue or benign lesions. Since experimental Brk expression in nontransformed mammary epithelial cells enhances their mitogenic response to epidermal growth factor, it was important to determine the role Brk plays in the proliferation of breast carcinoma cells and validate it as a therapeutic target. We have used RNA interference to efficiently and specifically downregulate Brk protein levels in breast carcinoma cells, and determined that this results in a significant suppression of their proliferation. Additionally, through the expression of a kinase-inactive mutant, we have determined that Brk can mediate promotion of proliferation via a kinase-independent mechanism, potentially functioning as an 'adapter'. These data identify Brk as a novel target for antiproliferative therapy in the majority of breast cancers, and illustrate the power of RNA interference for rapidly validating candidate therapeutic targets. PMID- 12902984 TI - Estrogen receptor (ER) beta1 and ERbetacx/beta2 inhibit ERalpha function differently in breast cancer cell line MCF7. AB - Estrogen receptor (ER) alpha plays an important role in the proliferation and progression of breast cancer. In order to explore the function of wild-type ERbeta (ERbeta1) and its variant form, ERbetacx/beta2, stable transformants of ERalpha-positive breast cancer MCF7 cells with ERbeta1 or ERbetacx/beta2 expression vector were established. Constitutive expression of ERbeta1 or ERbetacx/beta2 reduced the S phase population of the cell cycle in dish culture and the number of colonies in an anchorage-independent assay. DNA-protein complexes of ERE with nuclear extracts from ERbeta1 transformants were observed in the electrophoretic mobility shift assay, while no complex was observed for ERbetacx/beta2 transformants. Reporter gene assay using estrogen-responsive element (ERE)-luciferase showed less responsiveness to estrogen in these transformants compared with parental cells. Endogenous mRNA expression of two known estrogen-responsive genes, cathepsin D and IGFBP4, was weakly induced by estrogen in ERbeta1 and ERbetacx/beta2 transformants compared with parental cells. A comprehensive gene expression analysis using our custom-made cDNA microarray showed that MCF7 and ERbeta1 transformants had a similar gene expression profile, whereas ERbetacx/beta2 showed a distinct profile from others. These results indicate that ERbeta1 and ERbetacx/beta2 inhibit ERalpha function differently in MCF7 cells. PMID- 12902985 TI - SPARC/osteonectin is a frequent target for aberrant methylation in pancreatic adenocarcinoma and a mediator of tumor-stromal interactions. AB - Deregulated expression of SPARC/osteonectin, a secreted glycoprotein with multiple biological functions, has been associated with the progression of various cancers. Using microarrays, we previously identified SPARC as one of the genes induced by treatment with a DNA methylation inhibitor in pancreatic cancer cells. We therefore analysed the expression pattern and methylation status of the SPARC gene in pancreatic cancer. Gene expression profiling by oligonucleotide microarray and reverse transcription-PCR analyses demonstrated that SPARC mRNA was expressed in non-neoplastic pancreatic ductal epithelial cells, but was not expressed in a majority of pancreatic cancer cell lines. The loss of SPARC expression was associated with aberrant hypermethylation of its CpG island. Immunohistochemical labeling revealed that the SPARC protein was overexpressed in the stromal fibroblasts immediately adjacent to the neoplastic epithelium in primary pancreatic cancers, but rarely expressed in the cancers themselves. Primary fibroblasts derived from pancreatic cancer strongly expressed SPARC mRNA and secreted SPARC protein into the conditioned media, and treatment of pancreatic cancer cells with exogenous SPARC resulted in growth suppression. SPARC expression in fibroblasts from noncancerous pancreatic tissue was augmented by coculture with pancreatic cancer cells. These findings suggest that SPARC is a frequent target for aberrant methylation in pancreatic cancer and that SPARC expression in fibroblasts adjacent to pancreatic cancer cells is regulated through tumor-stromal interactions. PMID- 12902986 TI - PSF-TFE3 oncoprotein in papillary renal cell carcinoma inactivates TFE3 and p53 through cytoplasmic sequestration. AB - Papillary renal cell carcinomas are associated with chromosomal translocations involving the helix-loop-helix leucine-zipper region of the TFE3 gene on the X chromosome. These translocations lead to the expression of TFE3 chimeras of PRCC, RCC17, NonO and PSF (PTB-associated splicing factor). In this study, we explored the role of PSF-TFE3 fusion protein in mediating cell transformation. Unlike wild type TFE3 or PSF, which are nuclear proteins, PSF-TFE3 is not a nuclear protein and is targeted to the endosomal compartment. Although PSF-TFE3 has no effect on the nuclear localization of wild-type PSF, it sequesters wild-type TFE3 as well as p53 in the extranuclear compartment leading to functionally null p53 and TFE3 cells. In UOK-145 papillary renal carcinoma cells, which endogenously express PSF TFE3, siRNA complementary to the PSF-TFE3 fusion junction leads to a reduction in PSF-TFE3 and redistribution of endogenous TFE3 and p53 from the cytoplasmic compartment to the nucleus. Our results indicate that PSF-TFE3 acts through a novel mechanism, and exports TFE3, p53 and possibly other factors from the nucleus to the cytoplasm for degradation leading to the transformed phenotype. Thus, PSF-TFE3 is a promising target for the treatment for a subset of renal cell carcinomas. PMID- 12902987 TI - Role of the Fos family members, c-Fos, Fra-1 and Fra-2, in the regulation of cell motility. AB - The AP-1 transcription factor is composed of members of the Fos, Jun and ATF families, and plays a key role in tumor progression. We investigated whether Fos proteins regulate cell motility, and if so, whether this capacity is related to their transactivation potential. Two cell lines with different expression profiles of AP-1 were employed focusing on the Fos-family members c-Fos, Fra-1 and Fra-2. Transactivation motifs are found in c-Fos, but not in Fra-1 or Fra-2. The adenocarcinoma CSML0 cells display a low motility and do not express Fra-1 or Fra-2, and only very little c-Fos. In contrast, the fibroblastoid L929 cells express both Fra-1 and Fra-2, but no c-Fos, and these cells display a high motility. Transfection with Fra-1 or c-Fos, but not with Fra-2, strongly enhanced the motility of CSML0 cells. The effect of Fra-1 required the presence of the N terminal domain of this protein. Conversely, transfection with a Fos dominant negative mutant or with anti-sense fra-1 or fra-2, strongly reduced the motility of L929 cells. Changes in cell motility correlated with the morphological appearance and the degree of contact with the substratum. We conclude that Fos proteins have distinct roles in the regulation of cell motility. PMID- 12902988 TI - Both products of the mouse Ink4a/Arf locus suppress melanoma formation in vivo. AB - Deletion of the INK4a/ARF locus at 9p21 is detected with high frequency in human melanoma. Within a short genomic distance, this locus encodes several proteins with established tumor-suppressor roles in a broad spectrum of cancer types. Several lines of evidence support the view that p16INK4a and p19ARF exert the tumor-suppressor activities of this locus, although their relative importance in specific cancer types such as melanoma has been less rigorously documented on the genetic level. Here, we exploit a well-defined mouse model of RAS-induced melanomas to examine the impact of germline p16INK4a or p19ARF nullizygosity on melanoma formation. We demonstrate that loss of either Ink4a/Arf product can cooperate with RAS activation to produce clinically indistinguishable melanomas. In line with the common phenotypic end point, we further show that RAS+ p16INK4a /- melanomas sustain somatic inactivation of p19ARF-p53 and, correspondingly, that RAS+ p19ARF-/- melanomas experience high-frequency loss of p16INK4a. These genetic studies provide definitive proof that p16INK4a and p19ARF cooperate to suppress the development of melanoma in vivo. PMID- 12902990 TI - EDD, the human orthologue of the hyperplastic discs tumour suppressor gene, is amplified and overexpressed in cancer. AB - EDD (E3 isolated by differential display), located at chromosome 8q22.3, is the human orthologue of the Drosophila melanogaster tumour suppressor gene 'hyperplastic discs' and encodes a HECT domain E3 ubiquitin protein-ligase. To investigate the possible involvement of EDD in human cancer, several cancers from diverse tissue sites were analysed for allelic gain or loss (allelic imbalance, AI) at the EDD locus using an EDD-specific microsatellite, CEDD, and other polymorphic microsatellites mapped in the vicinity of the 8q22.3 locus. Of 143 cancers studied, 38 had AI at CEDD (42% of 90 informative cases). In 14 of these cases, discrete regions of imbalance encompassing 8q22.3 were present, while the remainder had more extensive 8q aberrations. AI of CEDD was most frequent in ovarian cancer (22/47 informative cases, 47%), particularly in the serous subtype (16/22, 73%), but was rare in benign and borderline ovarian tumours. AI was also common in breast cancer (31%), hepatocellular carcinoma (46%), squamous cell carcinoma of the tongue (50%) and metastatic melanoma (18%). AI is likely to represent amplification of the EDD gene locus rather than loss of heterozygosity, as quantitative RT-PCR and immunohistochemistry showed that EDD mRNA and protein are frequently overexpressed in breast and ovarian cancers, while among breast cancer cell lines EDD overexpression and increased gene copy number were correlated. These results demonstrate that AI at the EDD locus is common in a diversity of carcinomas and that the EDD gene is frequently overexpressed in breast and ovarian cancer, implying a potential role in cancer progression. PMID- 12902989 TI - Molecular cloning and characterization of LAPTM4B, a novel gene upregulated in hepatocellular carcinoma. AB - Lysosomal-associated protein transmembrane-4 beta (LAPTM4B), a novel gene upregulated in hepatocellular carcinoma (HCC), was cloned using fluorescence differential display, RACE, and RT-PCR. It contains seven exons and encodes a 35 kDa protein with four putative transmembrane regions. Both the N- and C-termini of the protein are proline-rich, and may serve as potential ligands for the SH3 domain. Immunohistochemical analysis localized the protein predominantly to intracellular membranes. Northern blot showed that the LAPTM4B mRNAs were remarkably upregulated in HCC (87.3%) and correlated inversely with differentiation status. LAPTM4B was also overexpressed in many HCC-derived cell lines. It was also highly expressed in fetal livers and certain adult normal tissues including the heart, skeletal muscle, testis, and ovary. Promoter function assays showed a distinct difference in the gene's activities between BEL7402 and HLE cell lines, suggesting that the transcription factors responsible for regulation of the gene in the two cell lines are different, and that possible negative regulatory cis-elements may exist upstream of the promoter region. It was demonstrated that the N-terminus of LAPTM4B was essential for survival of the cells. Cells harboring the full-length LAPTM4B cDNA expression clone displayed a slightly increased efficiency in colony formation. These results suggest that LAPTM4B is a potential protooncogene, whose overexpression is involved in carcinogenesis and progression of HCC. In normal cells, it may also play important roles such as regulation of cell proliferation and survival. PMID- 12902991 TI - Data mining the p53 pathway in the Fugu genome: evidence for strong conservation of the apoptotic pathway. AB - The p53 tumour suppressor gene belongs to a small family of related proteins that includes two other members, p63 and p73. Phylogenetic and functional studies suggest that p63 and p73 are ancient genes that have essential roles in normal development, whereas p53 seems to have evolved more recently to prevent cell transformation. In mammalian cells, a plethora of proteins have been found to specifically regulate p53 activity. The genome of the fish Fugu rubripes has been recently published. It is the second vertebrate genome for which the entire sequence is now available. Phylogenetic studies are essential in order to analyse and define signalling pathways important for cell cycle regulation. The presence or absence of a critical member in any pathway can shed light about the evolution of these pathways. The Fugu genome databank has been analysed for several members of the p53 network, including p53, p63 and p73. A good conservation of the network that regulates p53 stability and apoptosis has been found. We also discovered that some cofactors that cooperate with p53 for apoptosis are also well conserved and belong to multigene families not detected in the human genome. PMID- 12902992 TI - Effects of buprenorphine maintenance dose on mu-opioid receptor availability, plasma concentrations, and antagonist blockade in heroin-dependent volunteers. AB - The clinical effectiveness of opioid maintenance for heroin dependence is believed to result from a medication's ability to decrease mu-opioid receptor (muOR) availability thereby replacing agonist effects, alleviating withdrawal symptoms and attenuating heroin effects. We empirically tested this hypothesis in five heroin-dependent volunteers who were successively maintained on 32, 16, 2, and 0 mg daily buprenorphine (BUP) tablet doses. We predicted and confirmed that higher BUP doses would decrease in vivo muOR availability (measured with PET and [(11)C]carfentanil), increase plasma levels of BUP and its metabolite nor-BUP, and decrease withdrawal symptoms and hydromorphone (HYD) responses. Relative to placebo, BUP significantly decreased mean (+/-SEM) whole-brain muOR availability 41+/-8, 80+/-2, and 84+/-2% at 2, 16, and 32 mg, respectively. Regions of interest (ROIs) (prefrontal cortex, anterior cingulate, thalamus, amygdala, nucleus accumbens, caudate) showed similar dose-dependent effects. Changes in muOR availability varied across ROIs (prefrontal cortex, 47% vs amygdala, 27%) at BUP 2 mg, but were more homogeneous across ROIs at BUP 32 mg (94-98%; except thalamus, 88%). Relative to placebo (0 ng/ml), peak plasma levels of BUP and nor BUP were comparable and dose-dependent (0.5-1, 5-6, and 13-14 ng/ml at 2, 16, and 32 mg, respectively). muOR availability decreases were negatively correlated with BUP plasma level and positively correlated with questionnaire-based opioid withdrawal symptoms and attenuation of HYD symptoms. These findings suggest that high-dose BUP maintenance produces near-maximal muOR occupation, muOR availability correlates well with plasma levels, and BUP-related opioid symptoms and antagonist blockade exhibit concentration-effect relationships. PMID- 12902993 TI - SSR181507, a dopamine D2 receptor antagonist and 5-HT1A receptor agonist. II: Behavioral profile predictive of an atypical antipsychotic activity. AB - SSR181507 ((3-exo)-8-benzoyl-N-(((2S)7-chloro-2,3-dihydro-1,4-benzodioxin-1 yl)methyl)-8-azabicyclo(3.2.1)octane-3-methanamine monohydrochloride) is a novel tropanemethanamine benzodioxane that displays antagonist activity at dopamine D(2) receptors and agonist activity at 5-HT(1A) receptors. SSR181507 antagonized apomorphine-induced climbing in mice and stereotypies in rats (ED(50) of 2 and 3.4 mg/kg i.p., respectively) and blocked D-amphetamine-induced hyperlocomotion in rats at lower doses (0.3-1 mg/kg i.p.). At 1-10 mg/kg, it was found to disrupt active avoidance in mice. SSR181507 did not induce catalepsy in rats (MED>60 mg/kg i.p.) and antagonized (3-10 mg/kg i.p.) haloperidol-induced catalepsy. SSR181507 was also active in two models sensitive to antidepressant/anxiolytic drugs: in a guinea-pig pup/mother separation test, it decreased (1-3 mg/kg i.p.) the time spent vocalizing during the separation episode, and in a lithium-induced taste aversion procedure in rats, it partially reversed (3 mg/kg i.p.) the decrease of intake of a saccharin solution. Furthermore, SSR181507 increased (3 mg/kg i.p.) the latency time to paradoxical sleep in rats, an effect commonly observed with antidepressants. Coadministration of the selective 5-HT(1A) blocker SL88.0338 produced catalepsy and antagonized the effects of SSR181507 in the depression/anxiety tests, confirming the view that activation of 5-HT(1A) receptors confers an atypical profile on SSR181507, and is responsible for its antidepressant/anxiolytic properties. Finally, SSR181507 (1-3 mg/kg) did not affect memory performance in a Morris water maze task in rats. The pharmacological profile of SSR181507 suggests that it should control the symptoms of schizophrenia, in the absence of extrapyramidal signs and cognitive deficits, with the additional benefit of antidepressant/anxiolytic activities. PMID- 12902995 TI - The R7 subfamily of RGS proteins assists tachyphylaxis and acute tolerance at mu opioid receptors. AB - Members of the R7 subfamily of regulators of G-protein signaling (RGS) proteins (RGS6, RGS7, RGS9-2, and RGS11) are found in the mouse CNS. The expression of these proteins was effectively reduced in different neural structures by blocking their mRNA with antisense oligodeoxynucleotides (ODNs). This was achieved without noticeable changes in the binding characteristics of labeled beta-endorphin to opioid receptors. Knockdown of R7 proteins enhanced the potency of antinociception promoted by morphine and [D-Ala(2), N-MePhe(4), Gly-ol(5)] enkephalin (DAMGO)-both agonists at mu-opioid receptors. The duration of morphine analgesia was greatly increased in RGS9-2 and in RGS11 knockdown mice. The impairment of R7 proteins brought about different changes in the analgesic activity of selective delta agonists. Knockdown of RGS11 reduced [D Ala(2)]deltorphin II analgesic effects. Those of RGS6 and RGS9-2 proteins caused [D-Ala(2)]deltorphin II to produce a smoothened time-course curve-the peak effect blunted and analgesia extended during the declining phase. RGS9-2 impairment also promoted a similar pattern of change for [D-Pen(2,5)]-enkephalin (DPDPE). RGS7 deficient mice showed an increased response to both [D-Ala(2)]deltorphin II and DPDPE analgesic effects. A single intracerebroventricular (i.c.v.) ED(80) analgesic dose of morphine gave rise to acute tolerance in control mice, but did not promote tolerance in RGS6, RGS7, RGS9-2, or RGS11 knockdown animals. Thus, R7 proteins play a critical role in agonist tachyphylaxis and acute tolerance at mu opioid receptors, and show differences in their modulation of delta-opioid receptors. PMID- 12902996 TI - Neurocognitive performance and clinical changes in olanzapine-treated patients with schizophrenia. AB - Diffuse cognitive impairment characterizes patients with schizophrenia throughout the course of illness. The deficits persist despite clinical improvement associated with neuroleptic intervention and are related to outcome. It is unclear whether treatment with atypical agents is associated with improved cognition that relates to symptoms and outcome. Using a set of computerized neurocognitive measures, we evaluated whether the effects of olanzapine are greater than practice effects for specific neurocognitive domains that could provide targets for large-scale randomized studies. We enrolled 19 patients with schizophrenia before initiation of treatment with olanzapine and 16 of them were examined at 6 weeks and 6 months of follow-up. They were compared to 34 healthy participants who enrolled, 24 of whom were evaluated longitudinally. Improvement exceeding practice effects was observed in patients for abstraction and spatial memory and the latter correlated with clinical improvement in negative symptoms. These results suggest that some effects of olanzapine may impact both symptoms and cognitive performance. PMID- 12902994 TI - SSR181507, a dopamine D(2) receptor antagonist and 5-HT(1A) receptor agonist. I: Neurochemical and electrophysiological profile. AB - SSR181507 ((3-exo)-8-benzoyl-N-[[(2S)7-chloro-2,3-dihydro-1,4-benzodioxin-1 yl]methyl]-8-azabicyclo[3.2.1]octane-3-methanamine monohydrochloride) is a novel tropanemethanamine benzodioxane derivative that possesses high and selective affinities for D2-like and 5-HT(1A) receptors (K(I)=0.8, 0.2, and 0.2 nM for human D(2), D(3), and 5-HT(1A), respectively). In vivo, SSR181507 inhibited [(3)H]raclopride binding to D(2) receptors in the rat (ID(50)=0.9 and 1 mg/kg, i.p. in limbic system and striatum, respectively). It displayed D(2) antagonist and 5-HT(1A) agonist properties in the same concentration range in vitro (IC(50)=5.3 nM and EC(50)=2.3 nM, respectively, in the GTPgammaS model) and in the same dose range in vivo (ED(50)=1.6 and 0.7 mg/kg, i.p. on striatal DA and 5 HT synthesis, respectively, and 0.03-0.3 mg/kg, i.v. on dorsal raphe nucleus firing rate). It selectively enhanced Fos immunoreactivity in mesocorticolimbic areas as compared to the striatum. This regional selectivity was confirmed in electrophysiological studies where SSR181507, given acutely (0.1-3 mg/kg, i.p.) or chronically (3 mg/kg, i.p., o.d., 22 days), increased or decreased, respectively, the number of spontaneous active DA cells in the ventral tegmental area, but not in the substantia nigra. Moreover, SSR181507 increased both basal and phasic DA efflux (as assessed by microdialysis and electrochemistry) in the medial prefrontal cortex and nucleus accumbens, but not in the striatum. This study shows that the combination of D(2) receptor antagonism and 5-HT(1A) agonism, in the same dose range, confers on SSR181507 a unique neurochemical and electrophysiological profile and suggests the potential of this compound for the treatment of the main dimensions of schizophrenia. PMID- 12902997 TI - Individual differences in cocaine-induced locomotor activity in rats: behavioral characteristics, cocaine pharmacokinetics, and the dopamine transporter. AB - Outbred male Sprague-Dawley rats can be classified as either low or high cocaine responders (LCRs or HCRs, respectively) based on their locomotor response to acute cocaine. Concomitant measurement of dopamine clearance in these rats revealed that the differential behavioral responses are associated with the magnitude of dopamine transporter (DAT) inhibition by cocaine. Here, we investigated several factors that might contribute to cocaine-induced behavioral variability and its association with differential inhibition of DAT function. In rats classified as LCRs or HCRs after 10 mg/kg cocaine injection, we found no differences in (1) novelty-induced locomotion, (2) cocaine levels in dorsal striatum or nucleus accumbens (NAc), (3) DAT number or affinity in NAc, or (4) DAT affinity for cocaine in NAc. In rats given 20 mg/kg cocaine, behavior was more uniform across individuals, but still warranted separation into LCR/HCR categories. Additionally, we analyzed the stability of the LCR/HCR classification made during the first test with 10 or 20 mg/kg cocaine by retesting rats 7 days later with saline or cocaine (10 or 20 mg/kg). Before injection, HCRs were more active relative to LCRs and to their own behavior on the first test day. Following cocaine, LCRs and HCRs exhibited similar drug-induced changes in locomotion, but there were unique effects that depended on the cocaine dose given on the first and second test days. Our results argue against several likely explanations for individual differences in cocaine-induced behavior and highlight the influence of a single cocaine exposure on subsequent behavioral responses to the drug. PMID- 12902999 TI - Percutaneous arterial access: redefining the possibilities using suture-mediated closure (Perclose). AB - The standard for achieving post-procedure arteriotomy hemostasis has been 15 minute manual compression followed by 4- to 8-hour bed rest. Inadequate hemostasis can lead to local complications such as hematomas or significant complications requiring surgical intervention. Suture-mediated closure of the femoral arteriotomy offers an alternative to the standard manual compression to achieve secure hemostasis and has decreased overall access complications. Because the tissue is brought into complete apposition, the closure is independent of clot formation and the average time to hemostasis is significantly reduced. This article will discuss suture-mediated closure, the appropriate selection of patients and developing the proper technique for deployment. Using suture mediated closure will allow for a greater scope of eligible patients, no significant increase and often a decrease in complications, greater patient comfort and throughput, and utilization benefits for the practice of interventional radiology. PMID- 12903000 TI - Arterial puncture closure using a collagen plug, I. (Angio-Seal). AB - Closure devices can generally be categorized by their mechanism of obtaining hemostasis. Currently, devices are either suture-based, collagen-based, or utilize a combination of technologies. They can also be classified on the basis of whether or not they have an intravascular component or exclusively extra vascular components. The devices that are currently available differ markedly in their mechanism of action, deployment, advantages, and disadvantages. In this article, we will review in detail the Angio-Seal closure device, review some of the current literature regarding this device, and assess its complications, advantages, and disadvantages. PMID- 12903001 TI - Arterial puncture closure using a collagen plug, II. (VasoSeal). AB - Various vascular closure devices have come into common usage by most angio/interventional radiologists and cardiologists over the past 6 or 7 years. This has occurred despite the "real angiographers hold their own punctures" mindset that many of us learned in training. In our practice, we tried a number of different devices, but began using VasoSeal as our primary closure method in mid-1998. Since then, we have performed approximately 2400 procedures in which the VasoSeal device was deployed. The purpose of this article is to review the technique of VaseSeal closure of percutaneous arteriotomy sites. PMID- 12903002 TI - Arterial hemostasis using the Duett sealing device. AB - Using a liquid procoagulant, the Duett sealing device employs a distinctly different mechanism than other available devices for attaining hemostasis after femoral arterial puncture. Approved for use with 5 to 9 Fr sheath sizes, the Duett incorporates the typical vascular sheath in the closure procedure. Moreover, the liquid procoagulant, containing a mixture of thrombin and collagen, provides instant hemostasis and does not leave foreign material in the artery or in the subcutaneous tissues. Several studies, including one comparing the Duett to standard manual compression (SEAL), have shown the safety of the device as well as decreased times to ambulation. In addition, time to hemostasis was significantly decreased in patients receiving anticoagulation or antiplatelet therapy. The Duett sealing device is simple to use, safe, effective and a viable alternative to other available devices and/or manual compression. PMID- 12903003 TI - Non-invasive hemostatic closure devices: "patches and pads". AB - Initial device technologies developed to obtain hemostasis of percutaneous arteriotomy sites have used sutures or hemostatic plugs. Recently, a new class of noninvasive products have been introduced that promote rapid hemostasis in the form of a patch or pad. The purpose of this article is to review recent innovations using simple, but effective, hemostatic aids. PMID- 12903004 TI - Treatment of femoral artery pseudoaneurysms using ultrasound-guided thrombin injection. AB - Femoral artery pseudoaneurysms, a complication of femoral arterial puncture, is increasing in frequency with the proliferation of endovacular interventions. Pseudoaneurysms have historically been treated by open surgical repair, and more recently by ultrasound-guided compression. Ultrasound-guided compression is painful, and has a relatively low success rate of 51% to 73%. Since 1991, ultrasound-guided thrombin injection has become a treatment option. During injection of thrombin into a pseudoaneurysm, immediate thrombosis can be demonstrated within seconds. The entire procedure can be accomplished within 5 minutes, is simple to learn, and can be performed safely on an outpatient basis. PMID- 12903005 TI - Complications of arterial closure devices. AB - As of July 2000, there were 4 closure devices with Food and Drug Administration (FDA) approval. The devices are successfully deployed in 89% to 98% of cases, and the complication rates are comparable to those seen with manual compression. Minor access site complications are defined as bleeding not requiring transfusion or surgical repair, hematoma (<5 cm), and pain at the puncture site. Major complications include hematoma (>5 cm), bleeding at the site requiring transfusion, pseudoaneurysm, arteriovenous fistula, retroperitoneal hemorrhage, plug embolization, groin infection, and death. PMID- 12903007 TI - CTCAE v3.0: development of a comprehensive grading system for the adverse effects of cancer treatment. AB - Multiple systems have been developed for grading the adverse effects (AEs) of cancer treatment. The National Cancer Institute Common Toxicity Criteria (CTC) system has substantially evolved since its inception in 1983. The most recent version, CTCAE v3.0 (Common Terminology Criteria for Adverse Events version 3.0) represents the first comprehensive, multimodality grading system for reporting the acute and late effects of cancer treatment. The new CTC requires changes in the application of AE criteria including new guidelines regarding late effects, surgical and pediatric effects, multimodality issues, and for reporting the duration of an effect. It builds on the strengths of previous systems, represents a considerable effort among hundreds of participants, and signifies an international collaboration and consensus of the oncology research community. This article updates recent progress in the evolution of adverse effects grading systems and reviews the development of CTCAE v3.0. PMID- 12903008 TI - The biological basis of a comprehensive grading system for the adverse effects of cancer treatment. AB - As described in the previous article in this issue by Trotti et al, there have been major changes in the philosophy and scope of the new National Cancer Institute comprehensive grading system for treatment-related toxicities, Common Terminology Criteria for Adverse Events version 3.0 (CTCAE v3.0). The most prominent changes are the merging of early and late effects criteria into a single uniform document and the development of criteria that cover all treatment modalities. In this article, we briefly outline the biological support for the new grading system in the context of our current knowledge base. The clinical consequences of radiotherapy in normal tissue have been classically grouped temporally, into early and late effects, using a somewhat arbitrary dividing line, 90 days after commencement of radiotherapy. This definition was developed in an era of standard fractionation used alone or in simple sequential programs involving other modalities. However, most patients are now managed with multiple highly integrated modalities, often augmenting tissue injury and limiting our ability to ascribe any given effect to a particular modality. The use of complex concurrent or hybrid (concurrent/sequential) schedules also undermines the usefulness of a simplistic temporally defined early-late construct. Moreover, there is growing recognition that chemotherapy and surgery produce inherent long term biologic and clinical effects as well. Our basic understanding of the roles that surgery, chemotherapy, and radiation play in normal tissue response has expanded over the last decade because of vastly improved molecular techniques. The original biologic paradigm viewing acute and late tissue injury as a continuum of response and repair has been strengthened by these additional laboratory investigations. The expression of toxicity over time has been shown to be caused by a variety of cellular, tissue, environmental, and host factors. We continue to elucidate the roles of DNA damage, cytokines, chemokines, and associated inflammation, which lead in some cases to perpetuation of the wound healing response, progressive tissue fibrosis, and vascular compromise. The continuum model of tissue injury supports the recent changes in the common toxicity grading system. It also provides insights into potential targets and strategies for modulating response, which may in turn lead to effective interventions for altering the therapeutic ratio. PMID- 12903009 TI - Normal tissue effects: reporting and analysis. AB - Any effective cancer therapy developed to date is associated with a spectrum of normal tissue effects of varying incidence and severity. With an increasing number of novel therapeutic approaches undergoing clinical testing and an increased effort to optimize the established treatment modalities, methods for reliable quantification of normal tissue effects have become a key element in advancing cancer care. Here, we present a review of many of the issues involved in reporting and analyzing clinical normal tissue effect data. A distinction is introduced between explorative (science-driven) and pragmatic (patient-centered) studies. The desirable properties of criteria for reporting and grading toxicity are discussed from a biological and clinical perspective. Validation of toxicity criteria and the statistical issues involved in analyzing this type of data are presented with special emphasis on descriptors of the time evolution of toxicity. Finally, we discuss surrogate markers for late effects, mechanistic studies, and the design of clinical studies with normal tissue endpoints as a primary outcome. It is concluded that a consensus is required on guidelines for the reporting of normal tissue effects to improve the comparability of published reports on treatment outcome. PMID- 12903010 TI - Measuring postirradiation subcutaneous soft-tissue fibrosis: state-of-the-art and future directions. AB - Clinician-based rating scales and quantitative instruments used to quantify the severity of soft-tissue fibrosis were reviewed to determine if they were reliable and valid measures for clinical and research use. Existing clinician-based measures, the Medical Research Council (MRC) and European Organization for Research and Treatment of Cancer/Radiation Therapy Oncology Group (EORTC/RTOG), have preliminary testing of reliability and validity such that they provided a basis for the development of a revised measure. The revised measure, designed for use in National Cancer Institute-Common Toxicity Criteria for Adverse Events v3.0, evaluates subcutaneous and deep-tissue fibrosis separately and each is graded on a 5-point scale. The revised measure needs to be tested for reliability and validity, and it is recommended that it be used in concert with the MRC and/or EORTC/RTOG scales until such evidence is available and its utility can be determined. Quantitative measures, although attractive from a research perspective, will have limited clinical and multisite utility because of the apparatus required and the associated costs. Finally, a framework for evaluating fibrosis and the consequences of fibrosis is presented. PMID- 12903011 TI - The grading of lymphedema in oncology clinical trials. AB - Lymphedema is a common late toxicity of cancer therapy. This article describes the rationale and process utilized by the Lymphedema Working Group for the revision and expansion of the Common Toxicity Criteria version 2 (CTC v2.0) lymphedema criteria to produce the CTC v3.0 lymphedema criteria. Established clinician-based rating scales and quantitative instruments are reviewed in this article. None of the extant rating scales have been formally validated, nor has their reliability been assessed. Drawbacks of current scales were considered in formulating CTC v3.0 criteria. Most rely exclusively on volume to diagnose and grade lymphedema. This imposes significant clinical limitations, particularly in the assessment of toxicity in oncology clinical trials. Volume-based rating scales are of little value in rating the severity of bilateral limb and nonlimb edema. Problems with nonvolumetric staging systems (eg, CTC v2.0) include insufficient detail to permit useful discrimination of severity among the majority of lymphedema patients. Technologies for objectively quantifying lymphedema have been developed and validated. Although these are briefly reviewed, it is recognized that cost and access issues limit their widespread clinical utility and, as such, were not considered in developing the CTC v3.0 criteria. The CTC v3.0 lymphedema criteria adopted several innovations. Principle among these was the decision to generate separate criteria for volumetric increase, dermal changes, and subcutaneous fibrosis. We anticipate the use of the new CTC v3.0 lymphedema criteria to begin in mid-2003 for grading the key clinical features of this disorder in oncology clinical trials. The purpose of this article is to familiarize the reader with (1) background on the clinical features of lymphedema, (2) information on established lymphedema rating systems, (3) the consensus process and rationale of the Lymphedema Working Group, (4) the new CTC v3.0, and (5) quantitative techniques for assessment of lymphedema. PMID- 12903012 TI - How should we measure and report radiotherapy-induced xerostomia? AB - Xerostomia is commonly measured and graded using objective measures of major salivary gland output and observer-rated toxicity grading. The separation between the different grades is somewhat ambiguous in the current toxicity grading systems. We propose a new grading system based primarily on the functional deficits associated with xerostomia. Salivary flow rates have been added as a criterion to the grading system, notwithstanding the weak correlation reported in most studies between the symptoms and objective functional measures. In addition to the observer-rated toxicity grading, recording of patient-reported quality of life, using validated instruments, is encouraged. PMID- 12903013 TI - Quality of life in oncology trials: a clinical guide. AB - Over the last 2 decades, there has been an explosion of quality of life (QOL) studies in the oncology literature. The purpose of this review is to provide a basic guide regarding the state-of-the-art of QOL in oncology trials from a clinical perspective. This article aims to provide fundamental definitions, describe useful existing tools, and highlight the challenges in interpreting QOL studies. What exactly is QOL, and why is it important to measure in cancer trials? Key methodological issues will be addressed, including choosing the optimal instrument, basic psychometric properties, timing of QOL assessments, and statistical considerations, such as minimizing missing data. The critical issue of interpreting the clinical significance of QOL results will be explored, using both anchor- and distribution-based strategies. The Radiation Therapy Oncology Group (RTOG) model involving a triad of outcomes (clinical, humanistic/QOL, and economic) will also be described. The future challenge will be to translate QOL results into clinically meaningful interventions. PMID- 12903014 TI - Trends and advances in cancer survivorship research: challenge and opportunity. AB - With continued advances in strategies to detect cancer early and treat it effectively along with the aging of the population, the number of individuals living years beyond a cancer diagnosis can be expected to continue to increase. Although beneficial and often lifesaving against the diagnosed malignancy, most therapeutic modalities for cancer are associated with a spectrum of late complications ranging from minor and treatable to serious or, occasionally, potentially lethal. Taken as a whole, investigators conducting research among long-term cancer survivors (those 5 years or more beyond cancer diagnosis) are reporting that long-term adverse outcomes are more prevalent, serious, and persistent than expected in survivors of both pediatric and adult cancer. However, the long-term and late effects of cancer and its treatment remain poorly documented and understood among those diagnosed as adults. These data underscore the need for continued research in this ever-growing portion of the cancer survivorship spectrum. This article examines (1) definitional issues relevant to cancer survivorship, (2) the evolving paradigm of cancer survivorship research, (3) prevalence data for cancer survivors, (4) research needs and issues of particular relevance to long-term cancer survivors, and (5) cancer survivorship as a scientific research area with an overview of physiologic/medical sequelae of cancer diagnosis and treatment and the grading of late effects. A large and growing community of cancer survivors is one of the major achievements of cancer research over the past 3 decades. Both length and quality of survival are important endpoints. Many cancer survivors are at risk for and develop physiologic and psychosocial late and long-term effects of cancer treatment that may lead to premature mortality and morbidity. Interventions-therapeutic and lifestyle-may carry the potential to treat or ameliorate these late effects and must be developed, examined, and disseminated if found effective. PMID- 12903015 TI - Mucositis: current management and investigations. AB - Oral and pharyngeal mucositis present significant management problems for patients undergoing radiation for head and neck cancers. The incidence and severity of these sequelae of radiation have increased with the use of altered fractionated schedules and concurrent chemotherapy. Directed therapies for the prevention of mucositis have yet to be established. Over the past several decades, therapies have been and continue to be designed that interfere with the causative factors of mucositis in an effort to diminish its incidence. These novel therapies protect normal mucosa either through direct radioprotection or manipulation of growth factors and cytokines that are involved in mucosal repopulation. Other therapies have tried to counter inflammation or infection with the belief that the insult from radiation is exacerbated by these normal response mechanisms to injury. It has become clear that the quality of life of patients and the economic costs of supportive therapies needed because of toxicities of cancer therapies are significant concerns. Research leading to a better understanding of the mechanisms of mucosal injury will ideally result in more specific, more targeted, and effective strategies for prevention of mucositis rather than current management algorithms that rely primarily on costly symptom management. PMID- 12903016 TI - Late radiation-related fibrosis: pathogenesis, manifestations, and current management. AB - Radiation-induced fibrosis (RIF) represents one of the most common long-term adverse effects of curative radiotherapy. Current cancer treatment approaches, involving more intensive radiotherapy regimens, used in combination with systemic agents, will likely be associated with a higher incidence and greater degree of damage to normal tissues, especially RIF. Traditionally, the development of fibrosis after radiation therapy has been considered static and irreversible. Contemporary understanding recognizes RIF as a continuum of responses mediated by molecular pathways that may be amenable to interventions. Preliminary evidence suggests that pharmacological or other interventions may be possible to reverse the manifestation of the injury and restore function to tissues. A variety of strategies have been tested for the management of RIF, although formal trials of these therapies that permit treatment comparisons are unavailable at this time. It is critical that we formally evaluate new management approaches for RIF with larger patient accrual. To this end, it is also important to develop a means of registering its occurrence for outcome analysis and to refer these patients to colleagues familiar with optimal management and enrollment in clinical trials. PMID- 12903017 TI - Lymphedema management. AB - Lymphedema, defined as the abnormal accumulation of protein rich fluid dysfunction of the lymphatic system, is a common sequela of cancer therapy. The incidence is highest among patients who have undergone resection and irradiation of a lymph node bed. Recently, increased attention has been focused on the modification of anticancer therapies in an effort to minimize lymphatic compromise. Sentinel lymph node biopsy is an example of a surgical procedure developed to preserve lymphatic function. Concurrent with the development of less invasive treatments, the field of lymphedema management has evolved rapidly over the past decade. Combined manual therapy, often referred to as complex decongestive physiotherapy (CDP), has emerged as the standard of care. CDP combines compression bandaging, manual lymphatic drainage (a specialized massage technique), exercise, and skin care with extensive patient education. Case series collectively describing a mean 65% volume reduction in over 10,000 patients attest its efficacy. Pneumatic compression pumps were historically widely used to control lymphedema. Their use as an isolated treatment modality is now rare. Reliance on pumps diminished with the recognition that they may exacerbate truncal and genital lymphedema, as well as injure peripheral lymphatics when applied at high pressures. Many noncompressive approaches, particularly the use of benzopyrone medications and liposuction, continue to be used abroad. PMID- 12903018 TI - The prevention and treatment of radiotherapy - induced xerostomia. AB - Efforts to reduce the severity of postradiotherapy xerostomia include the use of salivary substitutes to gain symptomatic relief, salivary gland stimulants, agents delivered to protect the glands during radiotherapy (RT), and physical means to partially spare the major salivary glands from RT while adequately irradiating tumor targets. These means include advanced RT treatment planning and salivary tissue transfer to nonirradiated areas. The relative potential gain from each of these strategies is discussed in this article. The combination of partial salivary gland sparing and radiation protectors/stimulants may provide additive or synergistic gains in reducing the severity of xerostomia. PMID- 12903019 TI - Brain injury: current management and investigations. AB - Radiation, surgery, and chemotherapy, singly or combined, are modalities that are frequently used for the treatment of primary brain malignancies and metastases that secondarily spread to the brain. Each of the 3 modalities continues to improve with time; likewise the efficacy in management of malignant brain tumors has gradually improved. As cancer therapies become more effective with regard to disease control and survival, the potential impact of each modality on brain injury, either alone or when combined, becomes even more significant. In addition, with increasing survival, the likelihood of observing long-term injury is likely to increase. Thus, the expression of acute and long-term brain injury is becoming a more important factor in the management of patients with brain malignancies. This review presents current management and investigations associated with these modality-related injuries. PMID- 12903020 TI - Spinal cord and peripheral nerve injury: current management and investigations. AB - Treatment of patients having malignancy near or within the spinal cord can be more challenging or limited than in patients with brain tumors. This is largely because of the near total lack of noneloquent neural tissue associated with the spinal cord and/or intimately associated peripheral nerves. The adverse effects of surgery, radiation therapy, and chemotherapy or multimodality therapy can be acute or chronic, insidious, and often permanent in nature. Because cancer therapies (particularly combination therapies) have become more effective with regard to tumor control, the potential impact of each modality on spine and peripheral nerve injury has become even more significant. Similarly, with increasing survival, the likelihood of observing long-term injury is likely to increase. Thus, the expression of acute and long-term spine and peripheral nerve injury is becoming a more important factor in the management of patients with spine malignancies. This review presents current management and investigations associated with these modality-related injuries. PMID- 12903021 TI - Radiation-induced lung injury. AB - Radiation therapy (RT) for thoracic-region tumors often causes lung injury. The incidence of lung toxicity depends on the method of assessment (eg, radiographs, patient's symptoms, or functional endpoints such as pulmonary function tests). Three-dimensional (3D) treatment planning tools provide dosimetric predictors for the risk of symptomatic RT-induced lung injury and allow for beams to be selected to minimize these risks. A variety of cytokines have been implicated as indicators/mediators of lung injury. Recent work suggests that injury-associated tissue hypoxia perpetuates further injury. Sophisticated planning/delivery methods, such as intensity modulation, plus radioprotectors such as amifostine, hold promise to reduce the incidence of RT-induced lung injury. PMID- 12903022 TI - Radiation-associated cardiovascular disease: manifestations and management. AB - Irradiation of the heart incidental to the treatment of malignancies can cause a spectrum of cardiovascular complications. These include pericarditis, myocardial fibrosis, muscular dysfunction, valvular abnormalities, and conduction disturbances. Survivors of Hodgkin's disease and breast cancer survivors treated with radiotherapy after mastectomy appear to be the groups at highest risk for radiation-associated cardiovascular disease. Although modern techniques of chest radiotherapy have decreased its frequency by reducing the dose and volume of radiation exposure to the heart, survivors treated with radiation remain at increased risk of cardiovascular disease. The risk of fatal cardiovascular disease increases with younger age at treatment, longer follow-up, and higher dose volumes of exposure to the heart. Certain chemotherapeutic agents, such as anthracyclines, also increase the risk of damage to the heart. Cardiac damage associated with radiotherapy may be progressive. Screening of survivors may help identify those at highest risk for serious cardiovascular disease. The broad range of radiation-associated cardiovascular disease makes it necessary for survivors to be examined with multiple screening modalities, although data do not exist to support definitive recommendations on test frequency. PMID- 12903023 TI - Bowel injury: current and evolving management strategies. AB - The intestine is often dose limiting during abdominal and pelvic radiation therapy. Delayed bowel toxicity is difficult to manage and adversely impacts the quality of life of long-term cancer survivors. Of the 8 to 9 million cancer survivors currently living in the United States, more than half have had abdominal or pelvic tumors, and about 60% of these patients have undergone or will undergo radiation therapy. Therefore, interventions that limit postradiation intestinal dysfunction would significantly improve outcomes in a large number of patients. Worthwhile steps toward reducing toxicity of treatments have been taken recently by introducing dose-sculpting treatment techniques. However, prophylactic or therapeutic approaches derived from an improved understanding of the pathophysiology of bowel injury will result in further advances. This article reviews current principles in the diagnosis and management of intestinal radiation injury. It also provides an overview of investigational strategies aimed at reducing radiation-induced bowel toxicity. These strategies include free radical scavengers, antioxidants, cytoprotective agents, cytokines, and enterotrophic interventions, as well as modulators of intraluminal factors, endothelial dysfunction, and neuroimmune interactions. Preclinical testing in clinically relevant animal models will facilitate translation of these strategies into the clinic and contribute to improving cancer cure rates and quality of life in cancer survivors. PMID- 12903024 TI - Fatigue in the radiation therapy patient: current management and investigations. AB - Fatigue is one of the most common and distressing symptoms experienced by cancer patients. Research conducted over the past few years has documented that initiation of radiation therapy typically results in significant increases in fatigue severity. Preliminary evidence suggests that, among disease-free patients, this heightened fatigue remits to levels approaching those reported before initiation of treatment in the weeks or months after treatment completion. Potential sources of fatigue are varied and may include anemia, cytokine activation, psychological distress, concomitant symptoms and side effects, and concurrent medications. Efforts to manage fatigue in cancer patients should focus on correcting potential etiologies and relieving symptoms. For patients experiencing fatigue related to anemia, treatment with an erythropoietic agent should be considered. Preliminary evidence suggests that moderate exercise during radiation therapy may also be helpful in relieving fatigue. Additional research is needed to evaluate the effectiveness of these therapies as well as other intervention strategies, such as the use of antidepressant and psychostimulant medications. PMID- 12903032 TI - [Mitochondrial DNA mutations in patients with chronic progressive external ophthalmoplegia and Kearns-Sayre syndrome]. AB - OBJECTIVE: Kearns-Sayre syndrome (KSS) and chronic progressive external ophthalmoplegia (CPEO) belong to neurological diseases caused by a defect in the energy-producing system of mitochondria, and are known to be associated with a deletion in the mitochondrial genome. This study was aimed to understand with greater clearness the characteristics of mitochondrial DNA (mtDNA) mutations in 11 Chinese patients with CPEO (7 cases) or KSS (4 cases). METHODS: Densitometry of the bands on Southern blot, polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and sequencing were performed to search large scale deletions and A3243G point mutation in patients' muscle mtDNA. RESULTS: Large deletions in mtDNA were detected in 2 CPEO and 3 KSS patients, the size of deletion ranged from 3.0 kb to 8.0 kb. Moreover, mtDNA A3243G point mutation was identified in 1 KSS patient. The proportion of mutant mtDNA was 37.6%-87.0%. Direct sequencing of the PCR products revealed 5 novel large deletions not reported by others. CONCLUSION: The findings in this study being consistent with the reports by others, large scale deletions of mtDNA are frequently found in Chinese patients with KSS and CPEO. mtDNA A3243G mutation may also exist in some patients with KSS and CPEO. PMID- 12903033 TI - [Inherited coagulation factor VII deficiency caused by double heterozygotic mutations Arg304Gln and Arg304Trp]. AB - OBJECTIVE: To investigate the genotypes of mutations of an inherited coagulation factor VII(F VII) deficiency pedigree. METHODS: The diagnosis was validated by coagulant parameters. F VII gene mutations were analysed in the proband and her family members by DNA direct sequencing. The PCR fragments were cleaved by the Msp I restriction enzyme to confirm the mutations detected by sequencing was performed in this study. RESULTS: Double heterozygous mutations at the same coding site of amino acid were detected in propositus of the pedigree: a C to T mutation at position 11348 resulting in Arg304Trp substitution combined with a G to A mutation at position 11349 resulting in Arg304Gln substitution. Her farther had a G to A mutation at position 11349 and her mother had a C to T mutation at position 11348, respectively. Both were heterozygous mutations. One of her brothers had normal genotype, the other brother and all her three offsprings had heterozygous mutations. CONCLUSION: Double heterozygous mutations coding the same amino acid were found in a pedigree with hereditary coagulation factor VII deficiency. PMID- 12903034 TI - [Association between gene mutation of cytochrome P450 1A1 in exon 7 A4889G locus and susceptibility to endometriosis]. AB - OBJECTIVE: To assess the possible association between gene mutation of cytochrome P450 1A1(CYP1A1) in exon 7 A4889G locus and the susceptibility to endometriosis (EM). METHODS: Allele specific-polymerase chain reaction method was used to analyze gene mutation in exon 7 A4889G locus of CYP1A1 in 76 patients with endometriosis and 80 healthy controls. RESULTS: The frequency of allele G on A4889G locus of CYP1A1 gene showed a significant difference between the study cohort and the control group (Chi2=7.498, P<0.01), with an odds ratio of 1.957. Statistically significant difference in the frequencies of genotypes AA, AG and GG was observed between the two groups (Chi2=6.915, P<0.05). Individuals with homozygotes for G allele were at higher risk of suffering from EM when compared against those with homozygotes for A allele, the odds ratio being 3.437 (Chi2=5.430, P<0.05). CONCLUSION: The above results suggest that gene mutation of CYP1A1 in exon 7 A4889G locus might be a genetic susceptible factor of endometriosis. The mutation allele of CYP1A1 gene appears to increase the risk of endometriosis. PMID- 12903035 TI - [Study of functional L1 retrotransposon in human type 2 diabetes susceptibility loci]. AB - OBJECTIVE: To investigate the susceptibility gene of type 2 diabetes mellitus (T2DM) through a novel strategy. METHODS: Firstly, the common feature of the putative susceptibility genes in the reported susceptibility loci was searched by using NCBI BLAST, and a functional L1 retrotransposon in the loci was found. Secondly, the mRNA expression level of the functional L1 retrotransposon in 25 Han T2DM patients and 22 normal controls was investigated by reverse transcription-polymerase chain reaction, and statistical analysis was implemented in statistical package SPSS10.0. Thirdly, L1 retrotransponson genome mutation screening was performed via sequencing. RESULTS: Screening the human genome for the retrotransposon genome via alignment with the L1 genome using NCBI BLAST showed the functional L1 retrotransposons distribute on most chromosomes except for chromosomes 19, 21 and Y on which rare type 2 diabetes susceptibility loci were reported to reside, and their distribution sites are consistent with the locations of the reported candidate type 2 diabetes susceptibility loci. The mRNA expression level of the functional L1 retrotransposon in the T2DM patients was significantly lower than that in normal subjects (P<0.001). Nonsense mutations including deletion and/or point mutations were observed in all of the 6 T2DM patients tested, but no mutation was observed in all of the 4 normal controls tested. CONCLUSION: The functional L1 retrotransposon may be a candidate susceptibility gene of type 2 diabetes or a key regulator of the susceptibility genes, and it may be an ideal candidate biomarker for screening type 2 diabetes. PMID- 12903036 TI - [Sequence analysis and expression study of BRI gene in two lung adenocarcinoma cell lines with different metastatic potential]. AB - OBJECTIVE: To investigate the sequence of amyloid fibrils (BRI) gene and its expression in two lung adenocarcinoma cell lines AGZY83-a and Anip973 with the same tumor origin but different metastatic potential. METHODS: DNA sequencing, sequential G banding fluorescence in situ hybridization (FISH) and Northern blot were used to analyze the sequence and expression of BRI gene was in two lung adenocarcinoma cell lines with different metastatic potential. RESULTS: The expression of BRI gene was up-regulated in the highly metastatic cell line Anip973 and was down-regulated in the low metastatic cell line AGZY83-a from which the Anip973 was derived. FISH results disclosed that in the two cell lines, the same rearrangements existed in the chromosome region where BRI gene was located, but in Anip973, amplification took place in the chromosome region where BRI gene was located. DNA sequencing results showed different mutations in the 5' untranslated region of BRI gene in the two cell lines. CONCLUSION: The above results revealed that there was no relation between BRI gene differential expression and rearrangements of chromosome. The amplification of the chromosome region where BRI gene was located and the different mutations in the 5' untranslated region of BRI gene probably contributed to the differential expression. PMID- 12903037 TI - [Analysis on genetic polymorphism of 14 short tandem repeat loci on chromosome 7p14-15 and 12q13 in Chinese north Hans]. AB - OBJECTIVE: To analyze the genetic polymorphism of 6 short tandem repeat (STR) loci on chromosome 7p14-15 and 8 STR loci on chromosome 12q13 in Chinese north Hans. METHODS: Fluorescence-labeling polymerase chain reaction and capillary electrophoresis were used to analyze the genetic polymorphism of 100 randomly selected individuals from Chinese north Han nationality at 6 STR loci (D7S1808, D7S2250, D7S2251, D7S683, D7S656 and D7S528) on chromosome 7p14-15 and 8 STR loci(D12S1056, D12S1293, D12S83, D12S1655, D12S1662, D12S334, D12S137 and D12S102) on chromosome 12q13. RESULTS: In the Chinese north Han population, 7 alleles and 24 genotypes, 8 alleles and 27 genotypes, 7 alleles and 22 genotypes, 4 alleles and 10 genotypes, 6 alleles and 17 genotypes, 5 alleles and 13 genotypes were observed at D7S1808, D7S2250, D7S2251, D7S683, D7S656 and D7S528. The heterozygosities at the above 6 STR loci were 86%, 88%, 83%, 79%, 85% and 80%, respectively. Five alleles and 15 genotypes, 5 alleles and 15 genotypes, 8 alleles and 29 genotypes, 6 alleles and 17 genotypes, 6 alleles and 17 genotypes, 6 alleles and 19 genotypes, 5 alleles and 13 genotypes, 7 alleles and 24 genotypes were observed at D12S1056, D12S1293, D12S83, D12S1655, D12S1662, D12S334, D12S137 and D12S102. The heterozygosities at the above 8 STR loci were 86%, 84%, 87%, 82%, 84%, 85%, 81% and 89%, respectively. CONCLUSION: The distributions of allele frequencies of 6 STR loci on chromosome 7p14-15 and of 8 STR loci on chromosome 12q13 were consistent with the Hardy-Weinberg equilibrium. The highly genetic polymorphism was observed in Chinese north Han population. PMID- 12903038 TI - [Mutant thiopurine S-methyltransferase alleles among Jing Chinese in Guangxi province]. AB - OBJECTIVE: To investigate the mutant alleles of thiopurine S-methyltransferase (TPMT) among Jing Chinese. METHODS: Polymerse chain reaction-single strand conformation polymorphism (PCR-SSCP) techniques were developed for assaying exons 5, 7 and 10 of the TPMT gene respectively and were used to detect mutant TPMT alleles among Jing Chinese. RESULTS: Two cases of TPMT*3C (A719G) heterozygotes were identified in 103 Jing Chinese; other deleterious alleles such as TPMT*2 (G238C), TPMT*3A (G460A/A719G) and TPMT*3B (G460A) were not found; 27 cases of silent mutant allele TPMT*1S (T474C) were also identified (5 homozygotes and 22 heterozygotes). CONCLUSION: The PCR-SSCP assay established and adopted in this study was sensitive and reliable, which could be used to detect mutant TPMT alleles. Allele frequency of TPMT*3C is low among Jing Chinese (1.0%), and TPMT*3C appears to be the most prevalent deleterious allele in this population. PMID- 12903039 TI - [Study on the high-affinity IgE receptor beta gene polymorphism and its association with the susceptibility to allergic asthma in Han nationality of Hubei province]. AB - OBJECTIVE: To determine whether 2 polymorphism sites of the high-affinity IgE receptor beta (Fc epsilon RI beta) gene were associated with allergic asthma in Han nationality of Hubei province in China. METHODS: DNA and clinical data were obtained from allergic asthma patients and were compared with those from a group of healthy control subjects. The subjects were genotyped for the -109C/T and a coding variant Glu237Gly in Fc epsilon RI beta gene by polymerase chain reaction restriction fragment length polymorphism. RESULTS: (1) The genotype frequencies were 0.403 for -109T/T, 0.491 for -109T/C and 0.106 for -109C/C in allergic asthma patients in a Chinese population. No significant difference in the distribution of -109C/T polymorphism was found between allergic asthma patients and healthy control subjects; however, a homozygosity for the -109T allele was associated with increased total plasma IgE levels in patients with allergic asthma (F=7.523, P<0.01). (2) The frequency of Gly237Gly genotype was 0.051 in patient group, and 0.020 in control group. The allele frequencies of Gly237 in the patients and control were 0.236 and 0.136 respectively. There was a significant association between Gly/Gly genotype and allergic asthma. Among allergic asthma patients Gly237Gly was significantly associated with high IgE. CONCLUSION: These data suggested that the Gly237Gly genotype of the Fc epsilon RI beta gene conferred genetic susceptibility to allergic asthma in Chinese, which affected the total plasma IgE levels in the allergic asthma patients. And a homozygosity for the -109T allele was associated with increased total plasma IgE. PMID- 12903040 TI - [Studies on the relationship between D6S1581, a high frequency allele imbalance locus, and genetic susceptibility to nasopharyngeal carcinoma]. AB - OBJECTIVE: To investigate the relationship of nasopharyngeal carcinoma (NPC) with the high frequency allele imbalance locus D6S1581, and the NPC associated gene FBXO30 which is located near D6S1581. METHODS: Genescan was used to genotype D6S1581 of 12 NPC pedigrees, 85 sporadic NPC patients and 181 normal volunteers. Then parametric/nonparametric linkage analysis and association analysis were performed. RESULTS: D6S1581 was linked with NPC, a Lod score of 2.611436 (P=0.00245) was obtained, and a significant difference in allele frequency was observed between familial NPC and control (P<0.005). CONCLUSION: These results suggest that D6S1581 is highly associated with NPC, and there may be one or more NPC associated genes near D6S1581, including FBXO30. PMID- 12903041 TI - [Study on the distribution of vitamin D receptor gene start codon polymorphism in the Achangs and Hans]. AB - OBJECTIVE: To detect the difference between the Chinese Achang and Han ethnic groups in Yunnan province in the distribution of vitamin D receptor (VDR) gene start codon polymorphism. METHODS: Polymerase chain reaction-restriction fragment length polymorphism, gene sequencing and genetic analysis methods were used. A restriction fragment length polymorphism in the start codon of VDR (Fok I) gene was tested in the Achangs (n=68) and the Hans (n=92). RESULTS: The frequencies of FF, Ff and ff genotypes were found to be 18%, 35% and 47% in the Achangs, and 22%, 52% and 26% in the Hans, respectively. A significant difference was seen in the frequency distribution of VDR genotype between the Achangs and the Hans(Chi2=7.716, P=0.021). CONCLUSION: The Achang and Han ethnic groups differ in the frequency distribution of VDR gene start codon polymorphism. PMID- 12903042 TI - [The frequency distribution of flavin-containing monooxygenase 3 mutant alleles in 28 populations from Yunnan]. AB - OBJECTIVE: To study the frequency distribution of flavin-containing monooxygenase 3 (FMO3) mutant alleles in 28 populations originating from 24 ethnic minorities in Yunnan of China. METHODS: FMO3 genotypes were analyzed by polymerase chain reaction-restriction fragment length polymorphism. RESULTS: The average frequencies of FMO3/Stop(148), FMO3/Lys(158) and FMO3/Gly(308) were 0.395(0.174 0.803), 0.208 (0.056-0.414), 0.046(0-0.217), respectively. The frequencies of FMO3/Gly(308) in Blang, Huayaodai, Shuidai, Zhuang, De'ang, Jingpo, Nu and Hui populations were null. CONCLUSION: It was found that the frequencies of FMO3 mutant alleles varied not only in different ethnic groups, but also in different populations that stemmed from the same ethnic group. PMID- 12903044 TI - [Study on the genic polymorphism of HLA-DRB1 in a population in the north of China]. AB - OBJECTIVE: To investigate at the DNA level the polymorphism of HLA-DRB1 gene in Han population in Beijing of China. METHODS: This study was conducted in the Hans in Beijing. A total of 2810 normal and unrelated neonates were analyzed and subjected to HLA-DRB1 low resolution typing, and from among them 217 neonates were randomly selected for DRB1 high resolution typing by polymerase chain reaction-sequence specific primers technique. RESULTS: Fourteen HLA-DRB1 specificity and 39 alleles were found with HLA-DRB1 *09012, *1501, *07011, *08032 as the most common genes. Their gene frequencies and allelic distributions were compared to those observed in other ethnic populations. CONCLUSION: The results showed the characteristic of HLA-DRB1 distribution, and provided more comprehensive and accurate gene data that may serve as normal reference values for the population in Beijing, China. PMID- 12903043 TI - [Association analysis between attention-deficit hyperactivity disorder and Val158Met polymorphism of catechol-O-methyltransferase gene]. AB - OBJECTIVE: To investigate the association between attention-deficit hyperactivity disorder (ADHD) in Han Chinese children and Val158Met polymorphism of catechol-O methyltransferase (COMT) gene caused by the missense mutation of G158A in exon 4. METHODS: By using polymerase chain reaction-restriction fragment length polymorphisms the Val158Met polymorphism of COMT gene was tested in 117 children with the diagnosis of ADHD as defined by DSM-IV and in 105 healthy controls living in Shanghai. RESULTS: The frequencies of A allele were 25.21% and 23.81% in the ADHD group and the health controls respectively, which showed no significant difference between the two groups (Chi2=0.5197, P>0.05). There was also no significant difference in the distribution of all genotypes of COMT gene between the ADHD patients and the controls (P>0.05). CONCLUSION: It was suggested that for the Han Chinese children with ADHD in this study, there was no association between ADHD and Val158Met polymorphism of COMT gene. PMID- 12903045 TI - [Effect of 3' exonuclease activity of polymerase on extension of phosphorothioate modified primers]. AB - OBJECTIVE: To determine whether 3'phosphorothioate-modified-2 terminal mismatched primers can turn off DNA polymerization mediated by Exo(+) polymerase. METHODS: Two-directional primer extension was performed using polymerase with and without 3' exonuclease activity. The effects of unmodified primers and 3' phosphorothioate-modified primers on primer extension were evaluated. RESULTS: Exo(-) polymerase yielded products from matched and mismatched primers regardless of their modification. However, 3' phosphorothioate-modified primers with a single base mismatch at -2 position worked similarly to the terminal (-1) mismatched primers in triggering the novelly reported "off-switch" of Exo(+) polymerase. CONCLUSION: These data suggested that the "off-switch" can be of enormous application in the diagnosis of single gene diseases and in the association studies by single nucleotide polymorphism screening. PMID- 12903046 TI - [A dual-color fluorescence in situ hybridization study on the detection of inv(16) in acute myeloid leukemia]. AB - OBJECTIVE: To explore the value of dual-color fluorescence in situ hybridization (D-FISH) in the detection of inv(16) in acute myeloid leukemia (AML). METHODS: Eleven AML patients were investigated by D-FISH with two-color break apart probe for MYH11 labeled directly by fluorescein isocyanate (FITC) and a Texas Red. The results were associated or compared with those of cell morphology, cytogenetics, single color fluorescence in situ hybridization (FISH) and reverse transcription polymerase chain reaction (RT-PCR). RESULTS: Four cases (M4Eo three cases, M2a one case) had inv(16), of which one had trisomy 22 in addition to inv(16), while the other seven cases had no inv(16), of which, five cases (M4Eo three cases, M4 two cases)had a normal karyotype, one (M2a) had 5p+ and trisomy 22, one (M4Eo) had a translocation t(9;22) on G-banded karyotypic analysis. All 11 cases of AML were positive for the rearrangement of inv(16) detected by D-FISH. The average positive cell rate for these 11 AML patients was 93.45% (range 86.6%-98.7%). Of them, four had a minimal deletion of 16p13 in addition to inv(16). The results of D-FISH coincided with those of RT-PCR or single color FISH. CONCLUSION: D-FISH is a powerful tool for the detection of inv(16) due to its sensitivity and specificity. For raising the detecting rate of inv(16), it is necessary to screen inv(16) rearrangement by D-FISH in all M4- and M2-AML cases or the cases with trisomy 22, no matter whether they are accompanied by bone marrow eosinophilia. PMID- 12903047 TI - [Genome-wide scanning for susceptibility genes in researches on six HLA associated diseases]. AB - Six human leucocytic antigen(HLA)-associated diseases, including ankylosing spondylitis, rheumatoid arthritis, multiple sclerosis, systemic lupus erythematosus, type 1 diabetes mellitus and psoriasis, were selected as objects of this review. The characteristics of these diseases in whole-genome scans on susceptibility genes or loci undertaken to date were analyzed and compared. Meanwhile, the potential proposals for dealing with the existing problems were put forward. PMID- 12903048 TI - [Transmission disequilibrium test of polymorphisms of serotonin transporter gene and schizophrenia based on family trios]. AB - OBJECTIVE: To investigate the relationship between two polymorphisms (Intronic VNTR and 5-HTTLPR) of the serotonin transporter gene and schizophrenia. METHODS: A set of 314 schizophrenic trio samples collected from Shanghai, Xi'an and Jilin regions of China independently was subjected to analysis of the polymorphisms by transmission/disequilibrium test(TDT). RESULTS: No significantly preferential transmission of any allele was detected from both polymorphisms investigated. CONCLUSION: The results suggest that the serotonin transporter gene is unlikely to have a major contribution to susceptibility to schizophrenia in Han Chinese population. PMID- 12903049 TI - [Rapid diagnosis of non-deletion alpha-thalassemias by reverse dot blot]. AB - OBJECTIVE: To establish a rapid and convenient method of reverse dot blot (RDB) analysis for detecting the point mutations of non-deletion alpha-thalassemia in Chinese. METHODS: Label biotin to primers and amplify human alpha2 globin gene selectively, then hybridize PCR products with a set of oligonucleotide probes immobilized on strips, and develop colour to detect non-deletion alpha thalassemia defects. RESULTS: The PCR system using biotin-labeled primers could specifically amplify a 1085 bp fragment of human alpha2 globin gene which encompasses all six alpha-thalassemia mutations. After being hybridized with sequence-specific oligonucleotide probes and colour development, it could simultaneously identify all six types of non-deletion alpha-thalassemias encountered in Chinese. CONCLUSION: This method does not need semi-nested PCR, and the products amplified by biotinylated primers can be used directly to hybridize with the probes on strips under the identical conditions of hybridization. So, it is a specific and multiplex detection assay for screening non-deletion alpha-thalassemia defects in Chinese. PMID- 12903050 TI - [Expression of E-cadherin in human embryo]. AB - OBJECTIVE: To investigate the role of calcium dependent adhesion molecules (cadherin) during the implantation and development of human embryo by studying the expression of E-cadherin in human embryo and blastocysts. METHODS: Expression of E-cadherin in preimplantation embryos and blastocysts was detected by indirect immunofluorescence assay and quantitated by laser scanning confocal microscopy. RESULTS: Expression of E-cadherin was found in all the preimplantation embryos and blastocysts the present authors studied. The expression was higher in blastocysts than that in preimplantation embryos. CONCLUSION: The above results suggested that human embryos and blastocysts could express E-cadherin and the expression increased during their development. These may be of significance to the adhesion and implantation of the human embryo and blastocysts. PMID- 12903051 TI - [Detection of the gene-deleted female carriers of Duchenne/Becker muscular dystrophy using a fluorescent in situ hybridization based method]. AB - OBJECTIVE: To set up a fluorescent in situ hybridization (FISH) based method to detect the gene-deleted female carriers of Duchenne/Becker muscular dystrophy (DMD/BMD). METHODS: Multiplex polymerase chain reaction was used to identify the gene deletion DMD/BMD probands and their female relatives were checked by double color FISH. RESULTS: Two probands whose exon 46 of dystrophin gene was deleted, one had a positive pedigree and the other was a sporatic patient. In the case of the positive pedigree, four carriers were detected. In the case of the sporatic family, FISH showed that the mother of the proband was a somatic mosaicism. CONCLUSION: Combined with multiplex PCR, double-color FISH is a simple, fast, directly visual and accurate method. It is feasible to identify the carrier status of the female relatives of the gene deletion DMD/BMD probands. The detection of the somatic mosaicism is a prominent feature of FISH. PMID- 12903053 TI - [Establishment and preliminary application of screening methods for Y chromosome microdeletions in male infertility patients]. AB - OBJECTIVE: To develop a multiplex PCR protocol, which could be suitable for routine screening of microdeletions on the Y chromosome in azoospermic and oligozoospermic male infertility patients. METHODS: Five multiplex sets were established. Eighty-seven azoospermic and oligozoospermic patients undergoing intracytoplasmic sperm injection (ICSI) in the in vitro fertilization (IVF) center and 30 azoospermic men undergoing testicular biopsy in the clinic of Urology Surgery were screened for microdeletions of Y chromosome. RESULTS: A total of 19 (16.2%) cases of microdeletions were found in 117 azoospermic and oligozoospermic patients by screening of Y chromosome microdeletions. Of these, 11 cases (18.0%) were found in 61 oligozoospermic patients, and 8 cases (14.3%) were found in 56 azoospermic patients. CONCLUSION: The multiplex PCR protocol presented in this study is an easy-to-do and reliable method for detecting microdeletions on the Y chromosome. Routine screening of microdeletions on the Y chromosome for azoospermic and oligozoospermic patients is essential. PMID- 12903052 TI - [The association between glucocorticoid receptor gene G1666T polymorphism and cerebral infarction in Chinese]. AB - OBJECTIVE: To ascertain whether the human glucocorticoid receptor (GR) gene polymorphism is associated with cerebral infarction (CI) and essential hypertension (EH) in Chinese. METHODS: This is a case-control study of Chinese DNA samples abstracted from 163 subjects. The samples were analysed by a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) to determine the intron 4 variant of human GR gene. RESULTS: By analysis of the samples from both sexes as a whole, no association was observed between the EH+CI group and control group, and between the CI group and control group. However, by analysis of the samples from the females, weak association was observed between the EH+CI group and control group, and between the CI group and control group; the frequency of allele G was 0.64 for the EH+CI group, 0.69 for CI group and 0.46 for control group. CONCLUSION: The G allele may be a predisposing gene marker, GR gene intron 4 polymorphism contributes to the development of CI in females. PMID- 12903054 TI - [Evaluation of fetal DNA in maternal plasma for prenatal diagnosis]. AB - OBJECTIVE: To explore the application of fetal DNA in maternal plasma for uninvasive prenatal diagnosis. METHODS: The DNA template was extracted by hydroxybenzene-chloroform from 44 maternal (7-41 weeks) plasma. A Y-chromosome specific repeat sequence-DYZ1 gene (149 bp) was chosen to be amplified by PCR. RESULTS: The fragment was identified in all the plasma of male bearing pregnant women. The diagnostic accordance rate was 100.00%; two of the 22 female bearing pregnant women had false positive results. Among the 44 pregnant women, the diagnostic accordance rate was 88.89% at early pregnant stage, 100.00% at medium pregnant stage, and 96.55% at late stage respectively. The final accuracy of 95.45% was obtained in all cases. CONCLUSION: By means of hydroxybenzene chloroform extraction the authors of this article promoted the concentration and purity of the DNA template, and diagnosed more accurately. The results showed that free fetal DNA in the maternal plasma could be regarded as the gene resource for uninvasive prenatal diagnosis. PMID- 12903055 TI - [Genetic polymorphisms of short tandem repeats loci F13A01, FESFPS, vWA from Han population in Guizhou]. AB - OBJECTIVE: To understand the allele structure and genetic polymorphism at F13A01, FESFPS and vWA short tandem repeats (STRs) loci for Han population in Guizhou province of China, and to construct a preliminary database. METHODS: One hundred sixty unrelated individuals in Han population from Guizhou were analyzed by multiplex polymerase chain reaction. The resultant allele frequencies were compared with those of other population or race database. RESULTS: The genotype distributions of F13A01, FESFPS and vWA were in accordance with Hardy-Weinberg equilibrium. The combined discrimination power and polymorphism information content were 0.9984 and 0.9700 respectively. CONCLUSION: All of the three loci in this study provide useful markers for forensic paternity test and for genetic purposes. PMID- 12903056 TI - [The genetic characteristic of HLA-DRB1 locus in the Jiangsu-Zhejiang-Shanghai Han population and a comparison of its frequency distribution with that of other populations]. AB - OBJECTIVE: To investigate the genetic polymorphism of HLA-DRB1 locus in Jiangsu Zhejiang-Shanghai Han population and analyze the characteristic of the allele frequency distribution in comparison with that of other populations. METHODS: The technique of polymerase chain reaction-sequence specific primers (PCR-SSP) and reverse polymerase chain reaction-sequence specific oligonucleotide probe (PCR SSOP) was adopted in genotyping a sample of 626 unrelated healthy individuals collected from a Chinese Han population in Jiangsu-Zhejiang-Shanghai area. HLA DRB1*0101-1001, DRB3, DRB4 and DRB5 were detected. The allele frequency of HLA DRB1 was calculated, and the allele frequency distribution of HLA-DRB1 in this population was compared with the results from other populations. RESULTS: HLA DRB1*0101, 0301, 0701, 09012, 1001, 1201, 1202, 1301/02, 1303/04, 1401/04/05, 1402/03/1305, 1501/02, 16021 and 04xx, 08xx were detected in Jiangsu-Zhejiang Shanghai Han population. The common HLA-DRB1*allele included 09012(17.97%), 04xx(12.53%), 1202(11.42%) and 1501/02(11.02%). The polymorphism information content is 0.9024, and expected heterozygosity is 0.9634 in Jiangsu-Zhejiang Shanghai Han population. CONCLUSION: The HLA-DRB1 distribution of Jiangsu Zhejiang-Shanghai Han population shares some genetic characteristic with other Han populations, but it exhibits its own characteristic, suggesting the intermediate state of this population between the southern and northern Han populations. The polymorphism of HLA-DRB1 of Jiangsu-Zhejiang-Shanghai Han population is the most abundant one in this study. PMID- 12903057 TI - Branches of the splenic artery and splenic arterial segments. AB - In 200 adult autopsy specimens, the arterial supply to the pancreas and spleen was studied radiologically and by manual dissection. The splenic artery divided into two or three lobar arteries, which supplied its corresponding lobe; each lobar artery subsequently divided into two to four lobular branches. Six to twelve lobular branches were observed entering the splenic substance at the hilum. Lobar arteries did not anastomose with each other, hence, the lobes of the spleen are also termed segments. The lobules, however, were not found to be independent segments and the arteries of one lobule anastomosed with those of other lobules. The branching pattern of the splenic artery varied from one specimen to another, so much so that a prevailing pattern could not be identified. Polar arteries, particularly to the superior pole, arose quite proximal to the hilum in 51% of cases and were occasionally missed. In 45% of males and 40% of females, the posterior gastric artery arose from about the middle of the splenic artery. The splenic artery was not found to be tortuous in fetuses, newborns, and young children. Tortuosity was seen in only 10% of adults; thus, the characteristic tortuosity of the splenic artery appears to develop with age. PMID- 12903058 TI - Intramuscular innervation of the human soleus muscle: a 3D model. AB - The purpose of this study was to document the neural distribution patterns within the human soleus muscle using 3D computer modelling. Through serial dissection, pinning, and digitization, nerve distribution and muscle volume of a human cadaveric soleus muscle were documented and a detailed 3D computer model of neural distribution within the muscle volume was generated. Branching patterns demonstrated divisions that parallel architectural partitions within the soleus; that is, into anterior, posterior, and marginal soleus. Additionally, branching patterns demonstrated further partitioning of the posterior soleus into five distinct regions and the anterior soleus into two regions. Communication between nerve branches of the five regions of posterior soleus and between the anterior and posterior soleus were recorded. Knowledge of these anatomical partitions and their interaction is important as it will aid in the development of functional muscle models and in the understanding of normal and pathological muscle function. PMID- 12903059 TI - Unusual extended fibular origin of the human soleus muscle: possible morpho physiologic significance based on comparative anatomy. AB - A bilateral anomalous extended origin of the soleus muscle was observed in a 73 year-old female cadaver. It arose from the head, neck, and proximal two-thirds of the medial crest of the fibula and ran through the posterior intermuscular septum of the leg, from the lateral border of the fibula. The soleus muscle formed the vault of a muscular tunnel, overcoming the deep flexor muscles of the leg that was about 15 cm in length and directed inferiorly and laterally. The extended fibular origin delimited a blind recess lateral to the muscular tunnel and parallel to the posterior surface of the fibula. This recess measured 6.5 cm in length and extended 3.5 cm above the inferior opening of the muscular tunnel; the superior portion of the flexor hallucis longus was housed within it between the portions of the extended origin from the medial crest of the fibula and posterior intermuscular septum. The neurovascular bundle of the posterior leg coursed in the muscular tunnel. The tibial origin and calcaneal insertion of the soleus muscle were normal. Phylogenetic studies of the muscles of the lower limbs in mammals indicate that the fibular origin of soleus is more constant than the tibial origin and, in primates, the fibular origin is the only one observed in most monkeys. The case reported might be considered a conspicuous enlargement of the fibular origin observed in primates. This large fibular origin of the soleus muscle may prove to be a difficulty during surgery when accessing the proximal two-thirds of the fibula for ligation of the peroneal artery. PMID- 12903060 TI - Echocardiographic and morphologic examination of left ventricular false tendons in human and animal hearts. AB - False tendons are thin, fibrous or fibromuscular structures that traverse the cavity of the left ventricle with no connection to the valvular cusps; they may be single or multiple. We retrospectively analyzed echocardiograms for the prevalence of false tendons in the hearts of 368 (231 male, 137 female) newborns, infants, and children (mean age = 6.28 +/- 4.32 years) who were referred for echocardiography because of suspected acquired or congenital heart disease, but in whom no cardiac pathology was found. In addition, we studied the prevalence of false tendons in 90 hearts from three species of animals (dog, sheep, goat) and eight cadaveric human hearts. In our echocardiographic study, false tendons were detected in 97 of 368 hearts (26.4%). In our gross morphologic studies, false tendons were observed in most of the animal and human hearts: they were present in 5 of 8 (62.5%) human hearts, 14 of 20 (70%) dog hearts, 41 of 50 (82%) sheep hearts, and 16 of 20 (80%) goat hearts. The overall prevalence in animal hearts was 71 of 90 (78.8%). Histologic examination showed the false tendons to be composed of cardiac muscle, blood vessels, fibrous tissue, and Purkinje cells. The possible role of false tendons in innocent murmurs, cardiac rhythm disorders, or left ventricular dysfunction is discussed. PMID- 12903061 TI - Gender- and age-related differences in the morphology of the corpus callosum. AB - The size and shape of the adult corpus callosum may vary with gender and age. There is, however, little data on the morphology of the corpus callosum in the Indian population. The purpose of this study was to measure the size of the corpus callosum in normal adult Indian males and females, and to identify gender- and age-related differences. The size of the corpus callosum on midsagittal section was measured in 100 (50 males, 50 females) normal adult Indians using magnetic resonance imaging. The length of the corpus callosum, the width of the genu, trunk, and splenium, the area of the splenium, and the total area of the corpus callosum were measured. The length of the brain also was measured. Means were compared for significant difference by gender using the Student's unpaired t test and by age using ANOVA followed by Duncan's multiple range test. Gender was estimated by discriminant function analysis and age was estimated by regression analysis from significant parameters. The corpus callosum was longer in males and the discriminant score to differentiate gender was determined with an accuracy of 66%. The length of the corpus callosum increased with age and regression equations for predicting age was derived from the length of the corpus callosum. The width of the trunk and genu decreased with age in males but not in females. PMID- 12903062 TI - Clinical importance of ligamentous and osseous structures in the cervical uncovertebral foraminal region. AB - The vertebral artery, cervical spinal nerves, spinal nerve roots, and the bony and ligamentous tissue related to the cervical vertebrae are structures whose anatomy determines the path of a surgical approach. Defining the anatomy and, in particular, determining the precise location of vulnerable structures at the intervertebral foramen and the uncovertebral foraminal region (UVFR), a region defined by the uncinate process anteriorly, the facet joint posteriorly and the foramen transversarium laterally, has critical significance when selecting the safest surgical approach. We studied the anatomy of the vertebral artery, cervical spinal nerves, and spinal nerve roots within the UVFR in six cadaver specimens. We also obtained measurements of bony structures in 35 dry cervical vertebral columns, from C3-C7. The uncinate process (UP) projects superiorly from the posterolateral aspect of each cervical vertebral body, except for the first and second vertebrae. Because the posterior part of the UP lies adjacent to the vertebral artery, spinal nerve, and spinal nerve roots, its resection creates sufficient space to decompress these structures directly. The posterolateral surface of the UP is covered by ligamentous tissue that originates from the posterior longitudinal ligament and protects the neural and vascular structures during their decompression in the UVFR. PMID- 12903063 TI - High prevalence of tarsal coalitions and tarsal joint variants in a recent cadaver sample and its possible significance. AB - Tarsal coalitions (TC) are defined as fibrous (beyond normal ligaments), cartilaginous, or osseous unions of at least two tarsal bones. Most of the clinical studies report the prevalence of TC as <1%, but they disregard the asymptomatic coalitions. Because TC have been associated with pathologic conditions, including degenerative arthritic changes, knowledge of their prevalence has clinical importance. The aim of our study was to establish the prevalence of TC and tarsal joint variants. A total of 114 feet from 62 cadavers (average age = 78 years) without obvious foot pathologies were dissected at the Department of Anatomical Sciences, The University of Adelaide. Ten non-osseous TC in eight subjects were identified: two talocalcaneal and eight calcaneonavicular (occurred twice bilaterally). Variant calcaneonavicular and cuboideonavicular joints were found in 8% and 31% of feet, respectively. Other joint variants included a variable number of talocalcaneal joint surfaces and sesamoid bones. No secondary TC (due to trauma, infections, or neoplasm) were found. Our study demonstrated that the overall prevalence of TC is higher (13%) than previously thought; tarsal joint variations (39%) and sesamoid bones (42%) were common as well. The supposed secular increase in the prevalence of TC as well as the high number of anatomical variants could reflect a short-term response to altered life style or a microevolutionary trend due to relaxed selection. PMID- 12903064 TI - Collo-diaphysial angle of the femur in East African subjects. AB - The collo-diaphysial angle of East African subjects was investigated using 860 unilateral anteroposterior radiographs of the hip from 400 Ugandan and 460 Kenyan subjects, aged between 18 and 65 years using the standard techniques of Singh and Singh (1975). The angle ranged from 125-144 degrees with a mean of 133.3 degrees and a standard deviation (SD) of 5.3 degrees for Ugandan males; and 119-139 degrees, mean 123.4 degrees, SD 5.8 degrees for Ugandan females. The Kenyan values were 125-159 degrees, mean 140.2 degrees, SD 6.2 degrees for males; and 117-134 degrees, mean 125.7 degrees, SD 5.1 degrees for females, respectively. There were significant differences between males and females in both groups as well as between Ugandan and Kenyan males (P < 0.05). Using the demarking points method, only 19.0% of male Ugandans, 41.7% of male Kenyans, and 18.2% of female Kenyans could be identified by gender. This study documents for the first time the collo-diaphysial angle of East African subjects and has further confirmed that the angle is wider in men than women. PMID- 12903065 TI - Growth and aging of facial soft tissues: A computerized three-dimensional mesh diagram analysis. AB - The normal growth, development, and aging of facial soft tissues was studied by three-dimensional (3D) computerized mesh diagram analysis. The 3D coordinates of 50 soft-tissue landmarks were collected from 591 healthy white northern Italians (351 males, 240 females) 6-40 years of age. For each gender and age class, mean values were computed, and a standardized mesh of equidistant horizontal, vertical, and anterior-posterior lines was constructed. Within each age group, male meshes were superimposed on female meshes. For each gender, the 6-year-old reference mesh was superimposed on the reference mesh of each age group. The global (size plus shape) difference was evaluated by calculating the relevant displacement vectors for each landmark. Consequently, a size normalization was carried out and the shape difference was evaluated by calculating new relevant displacement vectors for each landmark. Growth and development were different along the three spatial planes: the largest increment was observed in the vertical dimension, with major modifications in the soft-tissue profile. The vertical dimension in males increased even after 30 years of age: ear dimensions increased, trichion moved superiorly and posteriorly, and pogonion, menton, and gonion moved anteriorly and inferiorly. In all age groups, size-standardized shape differences were found in the forehead, lower-third facial profile, eyes, cheeks, and ears. In each age class, male dimensions were larger than female dimensions. During childhood, gender differences in size were limited; shape differences were even less manifest. Overall, the profile was more anterior and inferior, the gonia were more inferior and more lateral, the forehead was more anterior, and the ears were larger in males than in females of corresponding age. PMID- 12903066 TI - Anatomical sciences in the allopathic medical school curriculum in the United States between 1967-2001. AB - The present article surveys the changes in the time allocated for the four disciplines in the anatomical sciences in the medical curricula of allopathic medical schools in the United States from 1967-2001. The results indicate that there was a great reduction in the laboratory portions, but not in the lecture portions of Gross Anatomy, Histology, Neuroanatomy, and Embryology. The greatest time reduction overall for the disciplines occurred between 1967-1973 (502 to 387 hr), followed by the time reduction between 1982-1995 (380 to 329 hr). PMID- 12903067 TI - Workshop in clinical anatomy for residents in gynecology and obstetrics. AB - A workshop in the clinical anatomy of the female pelvic viscera has been part of the training program for the first year residents in Gynecology and Obstetrics at the University of Padova since the 1999-2000 academic year. The purpose of the workshop is to offer a direct experience of practical anatomy despite a shortage of cadavers. It is designed for six residents who work in three teams on three specimens. The anatomical specimens are unembalmed, unfixed, female pelvic visceral blocs that are harvested from the cadavers 24 hr after death. They are stored at -12 degrees C and removed from the freezer 12 hr before the workshop. The workshop is 3 hr in length and has two parts: one on theory and one practical. In the theoretical section (30 min), the teacher presents the topographical anatomy of the specimens and the residents analyze a clinically oriented worksheet. In the practical section (2 hr), the residents identify viscera, vessels, and nerves through inspection and palpation, and then a step-by step dissection is carried out. In the last section (30 min), the workshop includes presentations on the three specimens that illustrate anatomical variability and assess knowledge of topographical anatomy. For many residents, this is the first practical experience of the anatomy of the female pelvic viscera because the anatomical courses for medical students do not include direct dissection by students. The unfixed viscera preserves the natural characteristics of the different tissues that exhibit the aspects of living organs, such as color, softness, and pliability, enhancing the knowledge of anatomy. The worksheet is structured as a guide to the anatomical basis of physical examination through inspection and palpation of the viscera. It also introduces the residents to the surgical anatomy of the female pelvis through a brief and selective dissection focused on the relationship between the different regional systems. PMID- 12903068 TI - Clinical anatomy of the fabella. PMID- 12903069 TI - Cranial deformation. PMID- 12903070 TI - Median and musculocutaneous nerves: variant formation and distribution. AB - An unusual formation of the median and musculocutaneous nerves was observed during routine dissection of the left upper limb of a 60-year-old Caucasian male cadaver. The median nerve was formed by the fusion of three roots, two from the lateral and one from the medial cord of the brachial plexus. The variant lateral root of the median nerve followed an anomalous course, crossing anterior to the distal part of the axillary artery. Moreover, in the distal half of the arm, the median nerve contributed a communicating branch to the musculocutaneous nerve. Injury to such a variant median nerve in the proximal arm may lead to paresthesia along the preaxial border of the forearm, weakness of elbow flexion, in addition to other manifestations of median nerve injury. The developmental and clinical significance of this anomaly is discussed. PMID- 12903074 TI - Value to basic surgical trainees of a combined post in accident and emergency medicine and anatomy demonstrating. PMID- 12903071 TI - Avulsion fracture of the pelvis: separation of the secondary ossification center in the superior margin of the acetabulum. AB - A 13-year-old female sustained a rare avulsion fracture of the secondary ossification center in the superior margin of the acetabulum as the result of contraction of the reflected head of the rectus femoris muscle. Diagnosis was made from plain films and CT scans with 3D image reconstruction. The patient was treated non-operatively by bed rest with semiflexion of the hip and knee, and appropriate analgesia. PMID- 12903075 TI - Tandem oligonucleotide synthesis using linker (First Base) phosphoramidites. AB - First Base linker phosphoramidites add the first nucleoside to underivatized solid-phase supports using a regular phosphoramidite coupling cycle. A cleavable 3'-O-ester linkage allows NH4OH treatment to rapidly generate products with only 3'-OH ends. Either single or multiple sequences linked in tandem can be prepared and no additional deprotection steps are required. PMID- 12903076 TI - Design and synthesis of modified nucleic acid systems. AB - Chemical modification of nucleic acids provides us with interesting and useful modified nucleic acid systems that have huge biochemical applications. We would like to discuss our preliminary results on modified functional oligonucleotides and modified antiviral nucleosides in the 29th Symposium on Nucleic Acids Chemistry. PMID- 12903077 TI - Design of intelligent nucleobases and DNA HOMO mapping. AB - We have designed a new nucleobase, benzodeazaadenine (BDA) that has a stronger charge transport ability than guanine and is not destroyed during charge transport process. By incorporating this new nucleobase into DNA, we demonstrated a protocol for real DNA nano-wire that is far superior to natural DNA. We also demonstrated that the selectivity for the interaction of Mn(II) ion with guanine N7 in G runs is a HOMO-controlled process, and as a consequence, the selectivity for G-metal ion interactions obtained by 15N-NMR studies would directly reflect the HOMO distribution of G-containing sequences in B-DNA. PMID- 12903078 TI - Cytidine protonation and base-pair intercalation in nucleic acid structures: the i-motif. PMID- 12903079 TI - Structure of DNA-RecA protein complex, intermediate of homologous recombination, determined by polarised-light spectroscopy. AB - The structure of long filament of DNA-RecA protein complex, active in homologous recombination, has been assessed using linear dichroism (LD) spectroscopy. The angular orientation of the DNA nucleobases relative to the fibre axis was estimated to be 70 degrees from the unique LD band of etheno-modified poly(dA) at 320 nm. Angular orientation data for 2 tryptophan and 7 tyrosine residues of RecA were deduced from differential LD of wild type RecA versus modified proteins which were engineered to attenuate the UV absorption of selected residues. The results revealed a rotation by some 40 degrees of the RecA subunits relative to the arrangement in crystal without DNA. Conformational changes are also indicated for tyrosine residues assigned to be involved in DNA binding or in RecA-RecA contacts. PMID- 12903080 TI - Chemical synthesis of novel taurine-containing uridine derivatives. AB - Recently, novel taurine-containing uridine derivatives were discovered in mammalian mitochondrial tRNAs, and these modified ribonucleosides existed at the first position of the anti-codon. This paper describes the chemical synthesis of these novel uridine derivatives, 5-taurinomethyluridine (tau m5U) and 5 taurinomethyl-2-thiouridine (tau m5s2U). These taurine-containing uridine derivatives were synthesized in the good yields by the reaction of the corresponding 5-hydroxymethyluridine derivatives with taurine under basic conditions. PMID- 12903081 TI - A novel C-C bond formation at the 5-position of uridine derivatives. AB - We have developed a novel C-C bond formation at the 5-position of uridine derivatives using intramolecular base-catalyzed Baylis-Hillman type reaction. PMID- 12903082 TI - Synthesis of carbocyclic 5'-nor uridine and cytidine derivatives. AB - Carbocyclic 5'-nor uridine and cytidine derivatives were synthesized. PMID- 12903083 TI - Ring opening of nucleosidic 1',2'-epoxides with aluminum reagent: a new stereoselective entry to 1'-C-substituted ribonucleosides. AB - Dimethyldioxirane-mediated epoxidation of 3',5'-O-(di-t-butylsilylene)-1',2' dehydro uridine (6) proceeded face-selectively to give 1',2'-alpha-epoxyuracil nucleoside (7). When 7 was treated with Me3Al, an anomeric mixture of the target 1'-C-alpha-methyluridine derivative 8 and its epimer 9 (86%, 8/9 = 5/1) was obtained. On the other hand, N3-protection improved the stereoselectivity in the ring-opening reaction. Thus, N3-benzyloxymethyl (BOM)-1',2'-epoxy nucleoside 11 obtained from 10 gave the target 12 as the sole product. PMID- 12903084 TI - Stereoselective synthesis of [5'-2H1][5'-13C]nucleosides. AB - An approach to a synthesis of stereoselective 5'-monodeuterated nucleoside combined with 13C label is described. A preparation of (5R)-D-[5-(2)H1;5 (13)C]ribose derivative of 1,2:5,6-di-O-isopropyliden-alpha-D-allofranose was successfully achieved by a 13C Wittig reaction using Ph3P13CH3I-BuLi to 5 oxoribose derivative and subsequent transformation into D-[5-(13)C]ribose derivative with an AD reaction, selective acylation, oxidation with NaIO4, and a stereoselective deuteride transfer reaction from Alpine-Borane-d to 5-oxo-D-[5 (13)C]ribose derivative as the main reaction. (5'R)-[5'-(2)H1;5' (13)C]nucleosides were synthesized in the established manner from this 5 (13)C/2H1-double-labeled ribose. PMID- 12903085 TI - Chemo- and regio-selective modifications of nucleic acids by acetaldehyde and crotonaldehyde. AB - The reactions of guanine nucleosides and nucleotides with acetaldehyde or crotonaldehyde were significantly accelerated even under mild conditions by the presence of a basic amino acid such as arginine and lysine to form smoothly and selectively the corresponding cyclic 1,N2-propano adducts. PMID- 12903086 TI - Synthesis and biological activity of adenophostin derivatives as IP3 receptor ligands. AB - The three derivatives of adenophostin A, which were the C-glycosidic analogs 3a and 3b, and the 5'-phenyl analog 4, were designed and synthesized as efficient IP3 receptor ligands. PMID- 12903087 TI - Synthesis and properties of 5'-amino-2',4'-BNA modified oligonucleotides with P3' ->N5' phosphoramidate linkages. AB - 5'-Amino-2',4'-BNA, a novel analogue of BNA series compounds, was synthesized and its incorporation into oligonucleosides via P3'-->N5' phosphoramidate linkage was successfully achieved on a DNA automatic synthesizer. The 5'-amino-2',4'-BNA modified oligonucleotides were found to have a potent hybridizing affinity for the single stranded RNA and double stranded DNA complements, and also to show a good resistant ability against SVPDE. PMID- 12903088 TI - A new protecting group for 5'-hydroxyl function of nucleotides in oligonucleotide synthesis without acid treatment utilizing unique properties of tritylthio group. AB - New protecting groups having a 2-aminomethylbezoyl skeleton, in which the reactive amino functions were blocked further by a tritylthio-type protecting group, were developed for the protection of the 5'-hydroxyl function of nucleosides during the oligonucleotide synthesis. These benzoate-type protecting groups were designed to be removed via an intramolecular cyclization following the removal of the tritylthio-type protecting group under mild oxidative conditions using diluted aqueous iodine solution. The new protecting groups would enable us to synthesize oligonucleotides without using any acid treatment. Furthermore, they can reduce the reaction steps required for a chain elongation cycle from four to three because the deprotection of the 5'-hydroxyl protecting group and the oxidation of the phosphite intermediate can be performed simultaneously by a single aqueous iodine treatment. PMID- 12903089 TI - A new strategy for the synthesis of oligodeoxynucleotides in the phosphoramidite method without base protection via phosphite intermediates. AB - Deoxyadenosine and deoxycytidine have highly nucleophilic amino groups. Therefore, without base protection, the undesired N-phosphitylations of these amino groups occur in the previous phosphoramidite methods. Now, we report that these N-phosphitylations were considerably suppressed in our new coupling strategy using HOBt as a promoter capable of formation of phosphite-type intermediates. PMID- 12903090 TI - Mutagenesis targeted by triple-helix forming oligonucleotides containing a reactive nucleoside analogue. AB - Recently, we have demonstrated that 2-amino-6-vinypurine derivative (1) achieves triplex-forming cross-linking with high selectivity toward the cytosine of the G C target site. In this study, we have investigated the cross-linking as well as induction of point mutations with the TFO incorporating 1 to a target site in a shuttle vector plasmid that replicates in mammalian cells. It was revealed that the TFO bearing 1 introduced mutations at the site of cross-linking. These results have suggested that the selective cross-linking with 1 might be useful for development of new biotechnology for targeted-mutagenesis. PMID- 12903091 TI - Duplex and triplex formation of oligodeoxyribonucleotides containing nucleobase intercalater conjugates. AB - Novel nucleoside analogues constituting of 2'-deoxyinosine and aromatic rings, which are connected by short linker groups, were synthesized and incorporated into oligodeoxyribonucleotides (ODNs). ODNs containing the nucleoside analogues formed stable duplexes and triplexes with target nucleic acids. The stacking interaction between base residues in the target nucleic acids and the aromatic groups of the nucleoside analogues appears to be a major cause of stabilization. PMID- 12903092 TI - Selective formation of non-natural type triplexes containing TA interrupting sites with the TFO incorporating W-shape nucleic acid (WNA) analogs. AB - Novel nucleoside analogs have been desired for selective formation of non-natural type triplexes containing TA or CG interrupting sites. We have previously reported that the W-shaped nucleic acid (WNA) would be a useful skeleton to develop new base analog for the formation of non-natural triplexes of antiparallel motif. In this study, we have found that triplex forming oligonucleotide (TFO) incorporating the new WNA analog, WNA-beta T, formed more stable triplex than natural triplex with high selectivity to the TA site. It is also noted that the TFO containing WNA-beta T did not form aggregates at the physiological condition of K+. PMID- 12903093 TI - An unnatural base pair between imidazolin-2-one and 2-amino-6-(2-thienyl)purine in replication and transcription. AB - Nucleosides of imidazolin-2-one (designated by z) were designed and synthesized as pairing partners of 2-amino-6-(2-thienyl)purine (designated by s). Previously, we developed an unnatural base pair between s and pyridine-2-one (designated by y), and polymerases specifically incorporated the substrate of y into DNA and RNA opposite s in templates. Although s was efficiently incorporated opposite y, A was also incorporated opposite y with high efficiency. The replacement of y by z effectively improved the incorporation selectivity of s. The incorporation efficiency of A opposite z decreased, but the efficient incorporation of s opposite z was maintained as compared to that of the s-y pairing. PMID- 12903095 TI - RNA LEGO: magnesium-dependent assembly of RNA building blocks through loop-loop interactions. AB - We describe the construction of nano-molecular assemblies using RNA building blocks the human immunodeficiency virus type 1 (HIV-1) dimerization initiation site (DIS) RNA, that forms stable base pairing through a magnesium-dependent loop loop interaction ("kissing"). RNA building blocks containing two DIS or DIS-like hairpins connected by a two nucleotide linker self-assembled to form specific structures as observed by non-denaturing polyacrylamide gel electrophoresis (PAGE). Furthermore, observation of "real time" formation of the molecular assemblies by circular dichroism (CD) spectroscopy was attempted. PMID- 12903094 TI - Probing DNA sequences by anthraquinone-modified DNA that is immobilized on gold surface. AB - 2'-Anthraquinone-modified oligonucleotide (AQ-ODN) possessing disulfide terminus has been immobilized on the gold electrode surface. The AQ-ODN-modified electrode showed faster electron transferability in double-stranded form than that in single-stranded form. PMID- 12903096 TI - Development of novel whole-mount in situ hybridization (WISH). AB - We have designed a novel fluorescent PNA probe which can be rapidly introduced into the living cell without any complicated pretreatment. We could post synthetically incorporate amino acid derivatives having membrane permeability function into the PNA probe by utilizing key compound 1 which we have already developed. We show that the PNA probe will be a candidate for a rapidly penetrating and non-degrading probe for WISH. PMID- 12903097 TI - Synthesis of the antisense oligonucleotides carrying the modified 2-5A molecules at their 5'-termini and their properties. AB - The synthesis of 8-methyladenosine (1)-substituted 2-5A tetramers with hydroxyalkyl groups at the 5'-phosphates and the corresponding 2-5A-antisense chimeras is described. These oligonucleotides were synthesized by the phosphoramidite method with a DNA/RNA synthesizer. These 2-5A tetramers, 8 and 9, with hydroxyethyl and hydroxybutyl groups at their 5'-phosphates were more resistant to hydrolysis by alkaline phosphatase than those without the hydroxyalkyl groups. Incorporation of the hydroxyethyl group into the 2-5A tetramer and 2-5A-antisense chimera slightly reduced the abilities of their analogs to activate recombinant human RNase L, but the abilities of the 2-5A tetramer, 11, and the 2-5A-antisense chimera, 15, with the hydroxyethyl group and 1 returned to 80 and 50% relative to those of the un-modified oligonucleotides, 7 and 12, respectively. Furthermore, the enzyme activated by 2-5A-antisense chimera 15 cleaved the complementary RNA as efficiently as that activated by 2-5A antisense chimera 12 without the hydroxyethyl group and 1. Thus, the 2-5A antisense chimera carrying the hydroxyethyl group and 1 was found to be a good candidate for an antisense molecule. PMID- 12903098 TI - Crystallization and preliminary X-ray diffraction studies of HutP protein: an RNA binding protein that regulates the transcription of hut operon in Bacillus subtilis. AB - HutP is an RNA-binding protein and regulates the expression of the histidine utilization (hut) operon in Bacillus subtilis by binding to cis-acting regulatory sequences on hut mRNA. A single point mutant (Val51Ile), which has increased affinity for the regulatory sequences, were purified and crystallized by the hanging drop vapor diffusion method. The space group was P2(1)3 with unit cell dimension a = b = c = 95.60 A resolution, and contain two molecules in the asymmetric unit. A complete MAD data set of 3.0 A resolution were collected at the LIII edge of Hg. PMID- 12903099 TI - Unique quadruplex structures of d(GGA)4 (12-mer) and d(GGA)8 (24-mer)--direct evidence of the formation of non-canonical base pairs and structural comparison. AB - We have reported that d(GGA)4 (12-mer) folds into an intramolecular parallel quadruplex with a G:G:G:G tetrad and a G(:A):G(:A):G(:A):G heptad and that two quadruplexes form a dimer. Here we present the unique structure of d(GGA)8 (24 mer) under physiological conditions. G1-A12 and G13-A24 segments of d(GGA)8 fold into an intramolecular parallel quadruplex with the tetrad and heptad, respectively, and two quadruplexes stack each other in a head-to-head manner. The formation of non-canonical G:G and G:A base pairs was directly certified by observing spin-spin couplings across hydrogen bonds with the aid of 13C-, 15N labelling of DNA. It was noted that the monomeric architecture of d(GGA)8 resembles the dimeric architecture of d(GGA)4. Interestingly, the recent X-ray study shows that the telomere DNA also folds into the similar intramolecular parallel quadruplex as we have reported for GGA-repeat DNAs. PMID- 12903101 TI - The X-ray crystallographic study of long chain left handed Z-DNA and polyamine complex and structural chemistry study of effection by polyamine for the Z-DNA. AB - We cleared the stabilization mechanism of Z-DNA by using the molecular dynamics calculation and the grid map simulated annealing docking method. Then, we clarified Z-DNA was stabilized by the metal counter ions whose electrical charge and the number of electrons are too many and by polyamine to go into the minor groove. We succeeded in stabilizing Z-DNA of the long chain by using these methods, and succeeded in the crystallization and the data collection up to 1.7 A resolution. PMID- 12903100 TI - Crystal structure of d(GCGAAAGCT) containing parallel-stranded duplex with homo base pairs and anti-parallel duplex with Watson-Crick base pairs. AB - A DNA fragment d(GCGAAAGCT), known to adopt a stable mini-hairpin structure in solution, has been crystallized under a slightly acidic condition, and its crystal structure has been determined at 2.5A resolution. In the first half of the oligomer, the sequence CGAA forms a parallel duplex with another symmetry related half through the homo base pairs, C2:C2+ (semi-protonated between the Watson-Crick sites), G3:G3 (between the minor groove sites), A4:A4 (between the major groove sites) and A5:A5 (between the Watson-Crick sites). The second halves of the parallel duplex are split away in the different direction, and extended to form an anti-parallel B-form duplex with another second half. PMID- 12903102 TI - Binding of threading intercalator to nucleic acids: thermodynamic analyses. AB - The constant melting temperature upon the change in the oligonucleotide concentration showed that the oligonucleotide d(CGCTTTGCG) formed a intramolecular hairpin rather than a duplex with an internal loop. We examined the thermodynamic properties of the interaction between the threading intercalator, naphthalene diimides designated as NDI, and a series of hairpin oligonucleotides, d(CGCTnGCG) (n = 3, 4, 5, and 6) designated as CGCn. A complex with a molar ratio of 1:1 was formed between NDI and CGCn. Because the observed negative entropy change was unfavorable for the binding of NDI to CGCn, the binding was driven by the large negative enthalpy change. All of the thermodynamic parameters for the threading intercalation were not significantly changed by the difference in the loop length n. The stem region rather than the loop region of CGCn is mainly involved in the binding of NDI. PMID- 12903103 TI - The molecular dynamics thermocycler. A new approach to sample conformational space, as exemplified by the RNA hairpin. AB - The approach called the molecular dynamics thermocycler employs repeated cycles of heating and cooling of the model during the course of unrestrained in aqua MD simulation. It is validated by confronting results of three 1.6 ns simulations using different heating temperatures, starting from a de novo constructed 12-mer RNA hairpin model containing a UGAA loop vs. the set of NMR derived coordinates (PDB code 1AFX). The lowest average RMS deviation and best coherence with the experimentally delivered structures was identified in case of the molecular dynamics thermocycler using Tmax = 400 K. PMID- 12903104 TI - Investigation of DNA strand exchange mechanisms mediated by cationic polymers. AB - It was observed that conjugation of dextran for PLL provided PLL-g-Dex with new properties of enhancing and accelerating inter-nucleotide recognition (1, 2, 3). In this paper, we report the effects of cationic PLL-g-Dex copolymers on base pair formation of self-complementary octamer oligonucleotide by using NMR and CD analysis. NMR studies revealed that PLL-g-Dex copolymer did not affect hydrogen bondings between bases in DNA duplex. CD studies also supported inert nature of the copolymer upon hybrid structure. PMID- 12903105 TI - Solution structure of the DNA complexes with an alkyl-linked bis(benzimidazole) compound, 1,4-bis[5-(4-methyl-1-piperazinyl)-2-benzimidazolyl]butane. AB - Solution structures of the 1:1 complexes of an alkyl-linked bis(benzimidazole) compound (4; Figure 1(a)) with the self-complementary DNA duplexes, d(CGCGAATTCGCG)2 and d(CGCAAATTTGCG)2, have been studied by 1H NMR spectroscopy and molecular mechanics calculations. The intermolecular NOE data and the energy minimized geometries of the 1:1 complexes indicate that 4 binds in the minor groove of the 5'-GAATTCG and 5'-CAAATTT regions of the DNA duplexes. PMID- 12903106 TI - Effect of secondary structure of short double-stranded RNA on RNAi efficiency. AB - The dangling nucleotides are very important for the RNA interference (RNAi). We investigate the effect of the dangling end on both the thermodynamic property of the RNA helix and efficiency of gene regulation in RNAi pathway. The result indicates that the four dangling adenines, which are more stable than two or three dangling residues, is not suitable for the RNAi efficiency. PMID- 12903107 TI - The structure and function of tmRNA. AB - tmRNA has a dual function both as tRNA and mRNA and facilitates trans translation. We made various mutation in tRNA domain of tmRNA. Our mutation studies give progressive suggestions on the structure of tmRNA. PMID- 12903108 TI - Alpha-helix substitution: novel approach for the design of a new zinc finger peptides for AT-rich sequence. AB - A novel strategy for the design of a zinc finger peptide on the basis of alpha helix substitution has been demonstrated. Sp1HM is a helix-substituted mutant for the wild-type Sp1(zf123) and its alpha-helix of each finger is replaced by that of fingers 4-6 of CF2-II. The circular dichroism spectrum of Sp1HM suggests that Sp1HM has an ordered secondary structure similar to Sp1(zf123). From the analyses of the DNA binding affinity and specificity by gel mobility shift assay, it is clearly indicated that Sp1HM specifically binds to the AT-rich sequence (5'-GTA TAT ATA-3') with 3 nM dissociation constants. Moreover, the zinc finger peptides for the sequence alternating between the AT- and GC-rich subsites can also be created by the alpha-helix substitution. This strategy is evidently effective and is also more convenient than the phage display method. Consequently, our design method is widely applicable to creating zinc finger peptides with novel binding specificities. PMID- 12903110 TI - Direct monitoring of EcoRII restriction enzyme reactions on a 27 MHz quartz crystal microbalance. AB - EcoRII is a type IIe restriction endonuclease characterized by a highly cooperative reaction mechanism that depends on simultaneous binding of the dimeric enzyme molecule to two copies of its recognition site. Gaining insights about reaction property of EcoRII binding to its recognition site and effect of divalent metal ion on DNA cleavage process, a DNA-immobilized quartz crystal microbalance (QCM), which enables real-time monitoring both the binding of enzyme and the cleavage reaction on DNA strands as mass changes, was utilized. PMID- 12903109 TI - Application of peptide nucleic acid (PNA) for detection of transcription factors binding probes. AB - We designed novel PNA monomer units that could post-synthetically and quantitatively introduce photo-functionalized molecules into PNA oligomers. Designed photo-functionalized PNAs were applicable to wide fields such as genome based analysis. In this study, we applied a photo-functionalized PNA oligomers for the detection of transcription factors binding probes. PMID- 12903111 TI - N4,5-dimethylcytosine, a novel hypermodified base in DNA. AB - Successive enzymatic modification of DNA by two methyltransferases that recognize identical or overlapping sequences and normally target either 5- or N4-position of the same cytosine residue, can lead to the formation of a doubly methylated base--N4,5-dimethylcytosine. Implications of such enzymatic "permethylation" of cytosine on the structure and interactions of DNA are examined. PMID- 12903112 TI - Photo-regulation of transcription by RNA polymerase with azobenzene-tethered promoter. AB - Photo-regulation of transcription reaction by T7-RNA polymerase was investigated by introducing an azobenzene residue into the non-template strand of T7-promoter. It was found that transcription was strongly interfered under dark or visible light irradiation, whereas it efficiently proceeded after UV irradiation. With this photo-responsive promoter, on-off switching of transcription by light irradiation was achieved. This strategy was also effective for the photo regulation of SP6-RNA polymerase. PMID- 12903113 TI - tRNA recognition by CCA-adding enzyme. AB - The CCA-adding enzyme [ATP (CTP): tRNA nucleotidyltransferase (EC. 2.7.7.25)] catalyzes the addition of the conserved CCA sequence to the 3'-terminus of tRNAs. It is known that CCA-adding enzymes from all three kingdoms recognize the elbow region of L-shape tRNA formed by the tertiary interaction between T-loop and D loop. We here demonstrate that C. elegans mitochondrial CCA-adding enzyme has broad substrate specificity toward nematode mitochondrial tRNAs which lack entire T- or D-arms, by cloning and characterizing the recombinant protein. PMID- 12903114 TI - Quality control of translation through the kinetic discrimination of tRNAs in the network of aminoacyl-tRNA synthetases. AB - It is known that each aminoacyl-tRNA synthetase (aaRS) specifically recognizes its cognate tRNAs to ensure the correct translation of the genetic information. However, we had previously demonstrated that mammalian mitochondrial seryl-tRNA synthetase (mt SerRS) can markedly misacylate mitochondrial (mt) tRNA(Gln). To investigate extensively misacylation reactions in mammalian mitochondrion, we purified overall twenty-two mt tRNAs from bovine liver, and determined their misacylation activities using mt SerRS. In addition to tRNA(Gln), tRNA(Ala) and tRNA(Asn) showed weak but significant serylation activities, which raises the possibility that each mammalian mt aaRS can misacylate several non-cognate mt tRNAs in varying degrees, but translational fidelity might be maintained by kinetic discrimination of tRNAs in the network of aaRSs. PMID- 12903115 TI - Molecular recognition of threonine tRNA by threonyl-tRNA synthetase from an extreme thermophilic archaeon, Aeropyrum pernix K1. AB - To investigate the recognition sites of tRNA(Thr) for threonyl-tRNA synthetase (ThrRS) from an extreme thermophilic and aerobic archaeon, Aeropyrum pernix K1, threonylation experiments using various in vitro mutant transcripts of tRNA(Thr) were examined. The results indicated that A. pernix ThrRS did recognize the first three base pairs of acceptor stem in addition to the second and the third letters of anticodon of tRNA(Thr), in spite of its N-terminal truncated unique structure. Discriminator base was not involved in recognition by A. pernix ThRS. These determinants were confirmed by the identity switching experiments from the in vitro mutants of A. pernix tRNA(Pro) and tRNA(Asn). PMID- 12903116 TI - Enzymatic incorporation of chemically-modified nucleotides into DNAs. AB - Modified 2'-deoxyuridine triphosphates bearing proteinous amino acids at a C5 position were prepared, and their substrate properties were investigated for KOD Dash DNA polymerase during PCR. The modified dUTPs bearing histidyl, lysyl, glutaminyl or seryl group produced the aimed 108 nt PCR products in good yields. In contrast, the analog bearing glutamyl group did not work as a substrate for KOD Dash while the analog bearing aspartyl group gave the product in a low yield. Moreover, not only KOD Dash but also three other thermostable DNA polymerases were tested as catalysts by use of C5 modified dUTPs with two different types of linker arms. Both Pfu and Vent(exo-) were relatively tolerant for the modification at the C5 position as well as KOD Dash. PMID- 12903117 TI - Substrate recognition by the human MTH1 protein. AB - A nucleotide pool sanitizing enzyme, the human MTH1 protein, hydrolyzes 2-hydroxy dATP, 8-hydroxy-dATP, and 8-hydroxyd-GTP. To examine the substrate recognition mechanism of the MTH1 protein, ten nucleotide analogs (8-bromo-dATP, 8-bromod GTP, deoxyisoinosine triphosphate, 8-hydroxy-dITP, 2-aminopurine-deoxyriboside triphosphate, 2-amino-dATP, deoxyxanthosine triphosphate, deoxyoxanosine triphosphate, dITP, and dUTP) were incubated with the protein. Of these, the former five nucleotides were hydrolyzed with various efficiencies. This results suggests the importance of the anti/syn-conformation and the functional groups on the 2 and 6-positions of the purine ring. PMID- 12903118 TI - The role of deoxycytidyl transferase activity of yeast Rev1 protein in the bypass of abasic sites. AB - We have studied the mutagenic specificity of abasic sites using the yeast oligonucleotides transformation assay. We introduced oligonucleotide containing a natural abasic site and a tetrahydrofuran abasic site into Rev1 mutants, rev1AA, which contains mutations of Asp467 and Glu468 residues of Rev1p to Ala in order to inactivate dCMP transferase activity, and rev1 delta, which lacks its whole coding sequence. The transformation efficiencies of rev1AA with abasic-containing oligonucleotides were lower than those of B7528, a strain proficient in REV1 gene, but much higher than rev1 delta mutant. Sequence analysis opposite the lesions showed that the mutation spectra were different among these three strains. PMID- 12903119 TI - Isolation of temperature sensitive mutants and their mapping in Agrobacterium tumefaciens. AB - The first temperature sensitive mutants of plant pathogenic bacterium Agrobacterium tumefaciens were isolated by Tn5 mutagenesis. Several genes responsible for mutants were identified and mapped on the circular and linear chromosomes in A. tumefaciens MAFF301001. PMID- 12903120 TI - Characterization of four ribosomal RNA operons in the genome of Agrobacterium tumefaciens MAFF301001. AB - The four ribosomal RNA (rrn) operons rrnA, rrnB, rrnC and were identified, sequenced completely and characterized individually rrnD in the whole genome of A tumefaciens MAFF301001. These rrn operons were located in the two topologically different chromosomal DNAs; rrnA and rrnD in the linear chromosome and rrnB and rrnC in the circular chromosome. Each operon coded for three ribosomal RNA subunit genes; 16S, 23S and 5S rRNA. The 16S-23S internal transcribed spacers (ITS) of the four rrn operons contain genes for tRNA-Ile and tRNA-Ala and tRNA Met downstream of 5S rDNA gene while the intergenic spacer between 23S rDNA and 5S rDNA lacked tRNA genes. Sequence alignment of 23S rRNAs of A. tumefaciens MAFF301001 and C58 strains showed unrelated sequences near the 5' end region suggesting the presence of intervening sequence (IVS). Primer extension analyses revealed that the primary transcription products for 16S and 23S rRNAs are 1497 and 2877 bases long, respectively. PMID- 12903121 TI - The role of nucleosome positioning in repression by the yeast alpha 2/Mcm1p repressor. AB - In Saccharomyces cerevisiae, alpha 2/Mcm1p represses a-cell specific genes. Previous studies indicated that repression by alpha 2/Mcm1p is associated with organization of positioned nucleosomes in promoters of a-cell specific genes such as STE6 and BAR1. Here, we examined the role of nucleosome positioning in repression of STE6-lacZ by alpha 2/Mcm1p. We showed that insertion of an A34 sequence, which is shown to disrupt an array of positioned nucleosomes, affects STE6-lacZ expression. Furthermore, artificial recruitment of TBP is facilitated by the insertion of A34 to derepress STE6-lacZ expression. These findings support the model that nucleosome positioning plays a regulatory role in repression by alpha 2/Mcm1p. PMID- 12903122 TI - Distamycin A, a minor groove binder, changes enediyne-induced DNA cleavage sites and enhances apoptosis. AB - We examined the effects of a minor groove binder, distamycin A, on DNA cleavage and apoptosis induced by an enediyne antitumor antibiotic, C1027, using 32P labeled DNA fragments obtained from human genes. C1027 alone induced DNA cleavage particularly at the 5'-TTTT-3'/3'-AAAA-5' sequence (cutting sites are underlined). The addition of distamycin A enhanced the double-strand DNA cleavage at the 5'-CCT-3'/3'-GGA-5' and 5'-CCA-3'/3'-GGT-5' sequences, with a two nucleotide 3'-stagger of the cleaved residues. These results suggest that distamycin A forms a heterodimer with C1027 to bind to DNA at GC-rich regions, resulting in amplification of DNA cleavage at these regions. Distamycin A enhanced C1027-induced DNA ladder formation and cytotoxicity in HL-60 cells. Therefore, amplification of DNA cleavage at GC-rich regions may result in enhancement of apoptosis. The present study on amplifiers of antitumor agents showed a novel approach to the potentially effective antitumor therapy. PMID- 12903123 TI - Transcriptional regulation of the c-jun gene by AP-1 repressor protein JDP2 during the differentiation of F9 cells. AB - ATF-2 and p300 cooperate in the activation of transcription of the c-jun gene during the differentiation of F9 cells. We show here that a repressor of AP-1, JDP2 (Jun dimerization protein 2), inhibits the transactivation of the c-jun gene by ATF-2 and p300, by recruitment of a histone deacetylase complex, thereby repressing the retinoic acid (RA)-induced transcription of the c-jun gene and inhibiting the RA-mediated differentiation of F9 cells. Furthermore, JDP2/HDAC complex was replaced by p300 complex on the DRE (differentiation response element) of the c-jun promoter within 24 h after the start of exposure of cells to RA, with an accompanying change in the histone-acetylation status of the chromatin, indicating that the initiation of transcription of the c-jun gene was controled by sequential action of HDAC(s) and HAT(s) on c-jun promoter. PMID- 12903124 TI - Down-regulation of VEGF mRNA expression by 2'-O,4'-C-ethylene-bridged nucleic acid (ENA) antisense oligonucleotides and investigation of non-target gene expression. AB - We studied the properties of 2'-O,4'-C-ethylene-bridged nucleic acid (ENA) oligonucleotides as antisense molecules. Compared to a phosphorothioate (PS) DNA and RNA heteroduplex, a duplex of an ENA/PS/ENA gapmer with RNA was a more effective substrate for RNase H-mediated cleavage. We designed ENA antisense oligonucleotides (AON) targeting human vascular endothelial growth factor (VEGF) mRNA. ENA AON with a cationic polymer considerably down-regulated VEGF mRNA expression, while no down-regulation by PS AON was observed. We also investigated the influence on the gene expression of genes other than VEGF by a gene database homology search and RT-PCR analysis. This method turned out to be an efficient approach to search for non-target genes sequence-specifically down-regulated by AON. In rat plasma, an ENA/PS/ENA gapmer was very stable after 24 hr, while a bridged nucleic acid (BNA) or locked nucleic acid (LNA) oligonucleotide and a PS oligonucleotide were half degraded in 4 hr. The high stability of ENA oligonucleotides in plasma would make ENA oligonucleotides ideal for in vivo studies. PMID- 12903126 TI - Study on the inhibition of protein production by L-cysteine derivatives of nucleic acid bases. AB - Isopoly (S-carboxymethyl-L-cysteine) derivatives of nucleic acids bases were prepared as antisense compounds. In previous studies, we investigated the properties of these compounds in vitro, and revealed that these compounds in vivo regulated the cell death presumably due to the inhibition of protein production. In this study, western and northern blots were carried out in order to reveal the mechanism of this inhibition for N-methyl-D-aspartate receptor in neuroblastoma x glioma hybrid NG108-15 cell line. In addition, we investigated the resistance of these compounds against cell extract and the metabolism. In conclusion, we proved that these compounds inhibited the protein production by antisense mechanism. PMID- 12903125 TI - Aminoacylation of tRNA by antisense molecule. AB - Aminoacylation of tRNA was attempted through formation of tRNA/DNA/aa-PNA (N aminoacylated peptide nucleic acid) ternary hybrid. A 23-mer DNA, that is complementary to a 3'-terminal of tRNA and to a 9-mer PNA carrying an amino acid unit, was designed to achieve close proximity between the amino acid and the 3' OH group of tRNA. The aminoacylation was carried out in a buffer solution containing imidazole. The aminoacylation was detected by nuclease S1 treatment followed by HPLC and MALDI-TOF MS. This novel methodology will open a way for easy and versatile aminoacylation of nonnatural amino acids onto specific tRNAs. PMID- 12903127 TI - Modulation of beta-secretase gene expression by action of catalytic nucleic acids. AB - Two sites of the initial translation sequence of beta-secretase (BACE) mRNA were chosen as targets for endogenously delivered hammerhead ribozymes. Ribozyme cassettes coding a catalytic core, flanked by the 5'-tRNA(Val) and the 3'-CTE (constitutive transport element) sequences, were cloned into a pUC119 plasmid and used for transfection of transformed primary human embryonal kidney (HEK293) cells. The ribozymes express in the test cells and inhibit beta-secretase gene biosynthesis on the level of mRNA (up to 95%) and on the level of protein (up to 90%) and thus may be considered as therapeutic inhibitors of beta-secretase activity. PMID- 12903128 TI - Effect of chemical features and stability of polyplexes on their gene transfer efficiency. AB - Chemically modified poly-L-lysine (PL) derivatives with two essential features, which we have recently reported on, have been used to study key factors affecting transgene expression efficiency. PL derivatives having both of N epsilon trimethyl lysine residue and 25 mole % serine residue showed enhanced transfection efficiency. When PL was modified in either way, no marked enhancement in gene expression was observed. These PL derivatives were found to be able to deliver plasmid DNA into nucleus of the transfected cells with a similar amount. Judging from the loss of EtBr fluorescence intercalating to DNA, the condensing tendency of the DNA were greatly affected by the sequence of the polypeptides used. This loose compaction seems to enhance the transgene expression because of an easy disassembly of the formed complexes or recognition of the DNA molecules in the complexes. PMID- 12903129 TI - Molecular design of hairpin polyamide possessing sequence-specific alkylation activity. AB - A novel type of sequence specific hairpin polyamide alkylating agents were synthesized and their alkylation activities were evaluated on linear 450 bp DNA fragment. To optimize the design of hairpin polyamide CPI-conjugates, several structural analogues were prepared. Analysis using high resolution denaturing gel electrophoresis suggests that such subtle structural differences effect the sequence specificity of the polyamides. PMID- 12903130 TI - Porphyrins and porphines inhibit the ribonuclease P reaction in vitro. AB - Porphyrin has been reported to bind to the T psi C stem of tRNA. This site is also recognized by ribonuclease P, which is essential and ubiquitous endoribonuclease responsible for the maturation of 5' ends of tRNA precursors. Thus, we investigated the effects of porphyrins on the in vitro reaction of ribonuclease P from Escherichia coli. The results showed that some of porphyrins inhibited the reaction more strongly than any other inhibitors reported so far. In addition to the benzimidazole inhibition that we have previously reported, these unusual substrate-binding inhibitions may provide new leads for the novel anti-bacterial reagent design. PMID- 12903131 TI - Development and application of siRNA expression vector. AB - RNA interference (RNAi) is a sequence-specific silencing phenomenon, which is induced by double-stranded RNA (dsRNA) and mediated through an evolutionarily conserved mechanism from plants to mammals. In mammalian cells, it has recently been reported that 21- or 22-nucleotide (nt) RNAs with 2-nt 3' overhangs (siRNA) induce RNAi without induction of the dsRNA-dependent inhibition of protein synthesis, known as the host defense system against viral infections. Moreover, we and other have developed siRNA expression systems utilizing a pol III promoter. Here we report a comparative analysis among various siRNA expression vectors and also demonstrate a regulatable RNAi in cells by using a tetracycline controlled U6 promoter. PMID- 12903132 TI - Construction of an allosteric trans-maxizyme targeting for two distinct oncogenes. AB - A maxizyme is dimmer of minimized ribozymes (minizymes) and can specifically cleave two target sites. The maxizyme also can allosterically cleave the target RNA only when it recognizes two target sites. In this study, for a cancer gene therapy, we focused two distinct oncogenes, cyclinD1 and hst-1, which are overexpressed in breast cancer cells. If we use conventional ribozymes for suppression of expression of those genes, these ribozymes affect not only these mRNAs in cancer cells but also those in normal cells because those genes are necessary for a growth factor-dependent signal transduction and a cell cycle in normal cells. To overcome this problem, we tried to design the trans-maxizyme that can cleave these mRNAs only in the breast cancer cells. PMID- 12903133 TI - Synthesis and application of bis (tert-butyl)-1-methyl pyrenyl-silyl chloride as a 5'-hydroxyl protecting group for nucleosides and as a fluorescent purification handle in oligonucleotide synthesis. AB - A fluorescently labelled monomer viz. 5'-O-1, 1-bis-(tert-butyl)-1'-methyl pyrenyl-silyl-thymidine-3'-phosphoramidite was synthesized. Different sequences were synthesized and were labelled using it and their fluorescence studies were carried out. PMID- 12903134 TI - Modeling of a possible evolutional process from a ribozyme to a catalytic RNP. AB - A model process for molecular evolution from an RNA enzyme to a catalytic RNA protein complex (RNP) is proposed. In the model, one RNA-RNA interaction in the enzyme is replaced by an RNA-protein interaction via an intermediary state where the original RNA-RNA and newly introduced RNA-protein interaction co-exist. To test the model, a catalytic RNP was designed and examined by employing the Tetrahymena ribozyme. PMID- 12903135 TI - Nonenzymatic DNA ligation in Escherichia coli cells. AB - We report on a new fluorescence reporting strategy in which dabsyl, a well-known quencher, activates a hydroxyl group in a probe to convert it to a leaving group. When a nucleophilic phosphorothioate probe binds adjacent to a dabsyl quenched probe, autoligation occurs, releasing the quencher, and lighting up the probes. These self-ligating DNA probes were used for sequence-specific detection of 16S rRNA in E. coli cells. Strong fluorescence was observed only when the phosphorothioate and quenched dabsyl probes bind side-by-side on a 16S rRNA target. The results demonstrate the use of QUAL probes to detect specific RNA sequences in bacterial cells without enzymes and without washing steps. PMID- 12903136 TI - Down regulation of human immunodeficiency virus type-1 (HIV-1) expression by Vif antisense RNA expression vectors in transfected cells. AB - The HIV-1 vif gene is a potential candidate in the quest for anti-retroviral interventions, due to its unique role in the target cell infection. We employed the antisense RNA strategy to determine the antiviral activity of intracellularly expressed anti-sense RNAs of various lengths complementary to the targeted HIV-1 vif gene. Expression vectors mediating the delivery of the vif-ORF, 5'-Vif, M vif, and 3'-vif antisense RNAs under the CMV promoter were constructed using pcDNA 3.1. The COS cells transfected with the antisense vectors showed a steady state of antisense RNA expression levels. In contrast, those co-transfected with the Infectious molecular clone, pNL-E, exhibited a significant reduction in the steady-state antisense RNA levels, which correlated with a significant reduction in p24 antigen production. Thus, this expression method for these antisense RNAs provides a promising gene therapy strategy for HIV-1. PMID- 12903137 TI - Patassium sensing oligonucleotide, PSO, based on DNA tetraplex formation. AB - Potassium Sensing Oligonucleotide, PSO, showing a high selectivity for K+ was synthesized by connecting 6-FAM and 6-TAMRA at the 5'- and 3'-termini of d(GGG TTA GGG TTA GGG TTA GGG), respectively. Circular dichroism (CD) spectra of PSO in the absence and presence of a various amount of KCl or NaCl revealed that PSO assumes an intramolecular or intermolecular antiparallel tetraplex structure in the presence of K+ and Na+, respectively. PMID- 12903138 TI - Recognition of DNA mismatch structures. AB - A dimeric form of 2-amino-1,8-naphthyridine strongly binds to a G-G mismatch by a zigzag intercalation. To gain insight into the mode of recognition for the G-G mismatch, we have synthesized a series of naphthyridine-heterocycle hybrids and evaluated the binding to G-G mismatches. Furthermore, assuming the G-G mismatch as a special variant of the G-X mismatch where X is G, A, and T, a general scheme for the zigzag binding of naphthyridine-heterocycle hybrids to a G-X mismatch was discussed. These structure activity relationships indicate that the zigzag binding of naphthyridine-heterocycle hybrids to a G-X mismatch could be deduced in a two-step sequence. PMID- 12903139 TI - Novel approach for SNP genotyping based on site-selective RNA scission. AB - Novel genotyping method for single nucleotide polymorphisms (SNPs), based on site selective RNA scission, has been developed. A substrate RNA is activated at two sites by complementary acridine-modified DNA having two acridine residues, and is site-selectively cleaved by metal ion catalyst to produce short RNA fragment containing the SNP site. Genotype of the substrate is accurately and easily determined by mass analysis of the fragment. PMID- 12903140 TI - Ion-channel sensors for electrochemical detection of DNA based on self-assembled PNA monolayers. AB - The gold electrodes modified with self-assembled monolayers composed of a 10-mer peptide nucleic acid (PNA) probe and 8-amino-1-octanethiol were used for the detection of the complementary oligonucleotide with a detection limit of 5.1 x 10(-10) M in a pH 7.0 phosphate buffer solution. In contrast, no response to a non-complementary oligonucleotide was observed. The electrode surface was positively charged in the phosphate buffer solution due to the protonated amine group of the thiol, where the electron transfer reaction between the electroactive marker [Ru(NH3)6]3+ and the electrode was hindered because of the electrostatic repulsion between them. Binding of the complementary oligonucleotide to the PNA probe monolayer cancels the positive charge at the electrode surface, and provides an excess negative charge at the surface, thereby facilitating the access of [Ru(NH3)6]3+ to the electrode surface and its redox reaction. PMID- 12903141 TI - Reaction of 4',5'-epoxynucleosides with carbon nucleophiles: stereoselective synthesis of 4'-C-carbon-substituted nucleosides. AB - 4',5'-Epoxythymidine (4) was obtained as a single diastereoisomer by oxidation of 3'-O-(t-butyldimethylsilyl)-4',5'-dehydrothymidine (3) with dimethyldioxirane. When the epoxide 4 was treated with allyltrimethylsilane in the presence of SnCl4, regio- (at the C4') and stereoselective (from the alpha-face) cleavage of the oxirane ring proceeded to afford a 4'-C-alpha-allylated derivative (5) as a sole product. In the reaction of 4 with trimethylaluminum, 4'-C-beta-methyl derivative (8) was obtained as a major product. PMID- 12903142 TI - Development of new amination reaction at 4-position of pyrimidine nucleosides. AB - A new method for the syntheses of the 4-amino-pyrimidine nucleosides (1a, b) has been developed. The method consists of conversion of uridines into quaternary ammonium intermediates (4a, b) by the reaction with p-toluenesulfonyl chloride(TsCl) in the presence of tertiary-amines, followed by amination with aq NH3. The method is expedient for large-scale preparation of cytidines like 2' deoxycytidine (1a) or 2'-deoxy-5-methylcytidine (1b). PMID- 12903143 TI - Discovery of a novel route to 2'-deoxy and 2'-functional pyrimidine nucleosides via 3',5'-O-sulfinyl xylo-nucleosides. AB - We synthesized 2,2'-anhydro-pyrimidine nucleosides through a novel and efficient rearrangement with 3',5'-O-sulfinyl xylo nucleosides which were prepared from an inexpensive starting material, D-xylose. A variety of 2'-deoxy and 2'-functional pyrimidine nucleosides can be prepared from 2,2'-anhydro-pyrimidine nucleosides. PMID- 12903144 TI - Synthesis and properties of N-t-butylaminoxyl nucleosides. AB - Novel purine nucleosides with a N-tert-butylhydroxylamino group at the 2-position of purine nucleus (1-3) were synthesized, which might prefer anti conformation and would not perturb DNA structure when incorporated into oligonucleotide. Oxidation of 1-3 by treatment with Ag2O or air afforded stable aminoxyl radicals (1'-3') and their properties were studied by EPR spectroscopy. 5-N-tert butylhydroxylamino uridine (4) was also synthesized. PMID- 12903146 TI - Design of a molecular beacon PNA. AB - We have designed a novel dual-labeled PNA oligomer, having both a fluorescent dye and a quencher, by utilizing key compounds 1 and 2. We showed that the designed dual-labeled PNA oligomer works as a molecular beacon PNA. We also investigated the optimization of a stem-loop structure which can supersensitize the function as a molecular beacon PNA. PMID- 12903145 TI - Synthesis of noraristeromycin analogues possessing SAH hydrolase inhibitory activity for the development of antimalaria agents. AB - Noraristeromycin analogues such as 3',4'-anhydronoraristeromycins, 2-amino-3',4' anhydronoraristeromycin and N6-cyclopropylnoraristeromycin were synthesized and their inhibitory activity against human and Plasmodium falciparum (P. falciparum) recombinant SAH hydrolase was investigated. Among them, 3',4' anhydronoraristeromycin showed inhibitory activity against human recombinant S adenosyl-L-homocysteine hydrolase (Ki = 12.4 microM, Kinact = 0.55 min-1). PMID- 12903147 TI - Synthesis of PNA using a Fmoc/Boc protecting group strategy. AB - Syntheses of PNA monomers whose nucleobases are protected with Boc group are described. To overcome the solubility problem of carboxymethyl-guanine derivative, we introduced 2-trimethylsilylethyl group at O6 as a second protecting group. PMID- 12903148 TI - Synthesis and properties of oligonucleotides containing novel 2',4'-BNA analogues (2',4'-BNACOC). AB - Preorganization of the nucleoside into proper conformation is one of the most promising approaches to develop the oligonucleotides strongly interacting with nucleic acid targets in a sequence-specific manner. We designed and synthesized the 2',4'-BNACOC monomer as a novel bridged nucleic acid analogue possessing a fixed N-type sugar conformation, and also successfully achieved its incorporation into oligonucleotides. The 2',4'-BNACOC modified oligonucleotides were found to have selective and strong binding-affinity for complementary RNA rather than DNA, and to show an excellent nuclease resistance ability. PMID- 12903149 TI - 2-Amino-6-vinyl purine as a tool for post synthetic conjugation of DNA with radio , spin-, fluorescence labels and peptides. AB - We have previously demonstrated that 2-amino-6-vinylpurine is an efficient cross linking agent with high selectivity towards cytidine. In this study, we applied this nucleobase analog to versatile conjugation of oligodeoxynucleotides with radio-, spin-, fluorescence- and peptide labels. The oligodeoxynucleotides incorporating 2-amino-6-vinylpurine nucleoside were reacted with amino- bearing nucleophies in an excess amount to give the corresponding conjugates in good yield. PMID- 12903150 TI - Novel base-labile protecting groups for 5'-hydroxyl protection in the synthesis of oligonucleotides. AB - The 6-(levulinyloxymethyl)-3-methoxy-2-nitrobenzoyl (LMM0NBz) and 2-(levulinyloxy methyl)-5-methoxy-4-nitrobenzoyl (LMMpNBz) groups were developed as novel base labile protection for the 5'-hydroxyl group in solid-phase oligonucleotide synthesis. A comparative study on the utility of LMMoNBz, LMMpNBz, and 2 (levulinyloxymethyl)-5-nitrobenzoyl (LMNBz) groups for the synthesis of oligodeoxyribonucleotides, and the synthesis of oligoribonucleotides using the LMMoNBz group in combination with the acid-labile 1-ethoxyethylk (EE) group to protect the 2'-hydroxyl group, are described. PMID- 12903151 TI - Reaction mechanism for the stereoselective internucleotidic bond formation by the oxazaphospholidine approach. AB - This paper describes the reaction mechanisms for the stereoselective synthesis of deoxyribnucleoside 3'-cyclic phosphoramidites as well as dinucleoside phosphite derivatives by the oxazaphospholidine approach. All the key reactions in the present approach were analyzed on the basis of ab initio molecular orbital calculations for the model compounds at the HF/6-31G* level. PMID- 12903152 TI - Conformation dependent DNA photoligation via sensitizer tethered 5 carboxyvinyluracil. AB - We report a novel template-directed reversible photoligation of oligodeoxynucleotides (ODNs) via sensitizer tethered 5-carboxyvinyluracil. In this system, control of photoligation and photosplitting at 366 nm was possible through with or without template ODN. PMID- 12903153 TI - RNA cleavage efficiency and catalytic turnover of an oligonucleotide-two copper complexes conjugate. AB - We synthesized a 2'-O-methyloligonucleotide with two terpyridine-Cu(II) complexes in the central part, and found that this antisense agent cleaves an RNA oligomer more efficiently than an analogous agent with one Cu(II) complex. Determination of the pH dependence of the activity revealed the existence of an optimum pH for the two complexes-agent, which may be a result of the cooperative action of the complexes. In addition, we demonstrated that the novel agent catalytically cleaved an excess of the RNA substrate. PMID- 12903154 TI - DNA recognition control of gamma-PRNA and mismatched base effects upon complex stability. AB - Oligomeric peptide ribonucleic acids (PRNA's), tethering 5'-amino-5' deoxyribonucleosides as a recognition site for nucleic acids, has been designed and synthesized by solid phase method. PRNA 12-mer found to form stable duplex with complementary DNA.PRNA.DNA duplex composed of PRNA with mismatched DNA has 10-degree lower melting temperature than fullmatched PRNA.DNA duplex. These results demonstrate that binding of PRNA with DNA is sequence specific. It should be emphasized that the on-off switching of PRNA.DNA duplex formation has been realized by the borate added as an external factor. PMID- 12903155 TI - Hybridization ability and base pair geometry of modified deoxycytidine derivatives having a 4-N-carbamoyl group. AB - Modified oligodeoxyribonucleotides incorporating 4-N-carbamoyldeoxycytidine derivatives were synthesized. The 1H-NMR of 4-N-carbamoyldeoxycytidine suggests that the carbamoyl group forms an intramolecular hydrogen bond with the cytosine ring nitrogen atom and this geometry would inhibit formation of the Watson-Crick base pair with a guanine base. However, the Tm analysis of the modified oligodeoxynucleotides revealed that, in the process of their hybridization with the complementary oligodeoxynucleotides, the orientation of the carbamoyl group changed in a manner where the stable Watson-Crick base pair can be formed with the guanine base. PMID- 12903156 TI - Anti-HIV-1 activity of L-DNA quadruplex. AB - Zintevir is a DNA 17mer that forms a quadruplex and shows strong anti-human immunodeficiency virus (HIV)-1 activity. The quadruplex formation is thought to be essential for the anti-HIV-1 activity of Zintevir. We synthesized the enantiomer of Zintevir and evaluated its structure and anti-HIV-1 activity. The results showed that the enantiomer has anti-HIV-1 activity comparable to that of Zintevir although it forms the mirror image quadruplex structure of Zintevir. PMID- 12903157 TI - Utility of Porous Glass with a new long-chain alkylamine spacer arm as a solid support for synthesis of oligodeoxyribonucleotides via the phosphoramidite method. AB - Porous Glass with an [[N-(2-aminoethyl)aminomethyl]phenyl]ethylsilyl spacer arm serving as a useful solid support in the synthesis of oligodeoxyribonucleotides via the phosphoramidite strategy. PMID- 12903158 TI - Control of photo-induced drug release by the use of conformational change of DNA. AB - Photo-induced drug release system which was controlled by triplet quenching using the molecular beacon strategy of photoreactive oligodeoxynucleotides (P-ODN) was developed. The strand ends of P-ODN were modified with a phenacyl ester of biotin and naphthalene as photoreactive group and triplet quencher, respectively. Photoirradiation to P-ODN in the presence of complementary DNA caused an efficient release of biotin. In contrast, the biotin release was suppressed in the absence of complementary DNA by the intramolecular triplet quenching in the stem-and-loop structure of P-ODN. PMID- 12903159 TI - Molecular sensing by pyrene excimer signal of fluorescent DNA aptamer. AB - The pyrene excimer label (2) has been incorporated into the adjacent residue to the binding site of anti-ATP DNA aptamer. The fluorescent aptamer exhibited the specific fluorescence signal only in the presence of ATP. PMID- 12903160 TI - Development of electrochemically gene-analyzing method using DNA-modified electrodes. AB - For electrochemical DNA sequencing, several redox-responsive uridines modified at C5 were synthesized and site-specifically incorporated into DNA. Self-assembled DNA monolayers were prepared on a gold electrode using the dsDNA containing modified uridines. By measuring differential pulse voltammetry (DPV) using DNA modified electrodes, the electrochemical response was strongly affected by the site of the redox-responsive uridine in dsDNA. PMID- 12903161 TI - Computational analysis of stacking interactions between 3-nitropyrrole and natural nucleobases. AB - The stacking energies between natural nucleobases and a universal base of 3 nitropyrrole (3-NP) were calculated by use of two theoretically independent quantum chemical methods, namely, molecular orbital (MO) and density function theory (DFT) calculations. The parameters required for molecular mechanics calculation of 3-NP were obtained by use of a software of Direct Force Field and used to evaluate the stacking energy of the complexes formed between 3-NP and canonical four nucleobases. Dependence of the twist angle between the two stacked bases on the stacking energy was studied in great detail. PMID- 12903162 TI - Molecular dynamics investigation of the double stranded oligonucleotide d(AT)6d(AT)6. AB - Molecular dynamics simulation of double stranded 12-meric oligonucleotide carrying an A and T alternating sequence, d(AT)6d(AT)6, over 1 ns was carried out under low NaCl conditions by using AMBER 6. It was found that the d(AT)6d(AT)6 was stabilized in the D-type structure which is an intermediate structure between A- and B-type ones and this result was in agreement with the previous report. PMID- 12903163 TI - Quadruplex structures of RNA 14-mer, r(GGAGGUUUUGGAGG) and DNA 14-mer, d(GGAGGTTTTGGAGG). AB - We have determined solution structure of r (GGAGGUUUUGGAGG) (R14) by NMR; the RNA 14-mer forms an intra-strand parallel quadruplex with a G-tetrad and a hexad, in which a G-tetrad core is augmented by association of two A residues. The quadruplex further forms a dimer through stacking interaction between the hexads. We also synthesized the corresponding DNA 14-mer, d (GGAGGTTTTGGAGG) (D14), and examined its properties and structure by CD, gel electrophoresis, and NMR. The CD and gel electrophoretic studies suggest that D14 forms a quadruplex entirely different from that of R14 in the presence of K+ ions; two molecules of D14 form a quadruplex containing antiparalle strands and diagonal loops. The NMR study also gave the results that are consistent with such structure: alternate glycosidic conformation, 5'G(syn)-G(anti)3', and characteristic chemical shift data. PMID- 12903164 TI - X-ray structure of d(CGAA), parallel-stranded DNA duplex with homo base pairs. AB - To clarify whether the parallel right-handed duplex found in the d(GCGAAAGCT) crystal is possible to occur without the anti-parallel part, the crystal structures of d(CGAA) at pH6.0 and pH7.0 have been determined by X-ray analysis. There are two independent duplexes in the crystallographic asymmetric unit. In each duplex, two chains are aligned in parallel fashion to form a right-handed double helix through the homo base pair formations, the sugar puckers being essentially C2'-endo. The C1:C1 pair is achieved by hemi-protonation. The G2:G2, A3:A3 and A4:A4 pairs occur between the minor groove sites, between the major groove sites and between the Watson-Crick sites, respectively. In every pairing, the two bases are located at the trans positions to each other. These structural features are the same as those at pH7.0, suggesting that the C1:C1 pair is hemi protonated even at the neutral condition. PMID- 12903165 TI - X-ray structure of d(GCGAAGC); switching of partner for G:A pair in duplex form. AB - Crystal structure of a DNA fragment d(GCGAAGC), known to adopt a stable mini hairpin structure in solution, has been determined at 1.6A resolution. Two heptamers are associated to form a duplex with a molecular two-fold symmetry. Three duplexes in the asymmetric unit have a similar structure. At the both ends of each duplexes, two Watson-Crick G:C pairs constitute the stem region. In the central part, two sheared pairs of G:A and A:G are formed, the two G bases being stacked as well as the two A bases. At this point, the two strands are crossed between the two base-stacked columns. The adenine moiety of the bulged A5 residue, which intercalates between the A4 and G6 residues, makes a small bending of the duplex at the two sites. The difference between the bulge-in structure of d(GCGAAGC) and the zipper-like duplex of d(GCGAAAGC) is ascribed to switching the partner of the sheared G:A pairs. PMID- 12903166 TI - Triplex formation involving 2',4'-BNA with isoquinolone base analogue: efficient and selective recognition of C:G interruption. AB - We examined the thermodynamic properties of 2',4'-bridged nucleic acid containing 1-isoquinolone as a nucleobase (QB) to recognize a C interruption in the homopurine strand of the target duplex for pyrimidine motif triplex formation at neutral pH. The triplex formation involving triplex-forming oligonucleotide with QB is highly sequence-selective to specifically recognize C:G target base pair rather than the other G:C, T:A, or A:T base pairs. QB.C:G triad gives significantly larger binding constant than T.C:G triad, which has been known to be the most stable combination in natural base.C:G triad. Our results certainly support the idea that QB could be a key nucleoside to recognize a C interruption in the homopurine strand of the target duplex with high binding affinity and selectivity, and reduce the restriction of target sequences for triplex formation. PMID- 12903167 TI - Binding analysis between naphthyridine dimers and GG mismatch DNA. AB - Kinetics and thermodynamics studies of the binding between naphthyridine dimer and DNA duplexes having a G-G mismatch sequence were performed by using Surface Plasmon Resonance (SPR) assay. Naphthyridine dimers possessing a different length of PEG linker were synthesized and immobilized to the dextran surface of the sensor chip to obtain an improved sensitivity of the SPR sensor toward a G-G mismatch. The effect of the directly neighboring base pair to the G-G mismatch was estimated from the thermodynamic parameters of the complex. These results indicate that stacking with adjacent base pair and naphthyridine dimer is the key roll for the formation of the complex between mismatched base pair and naphthyridine dimer. PMID- 12903168 TI - Physicochemical analysis of nucleobase-metal interactions. AB - It is important to know what kinds of chemical bonds are formed between metal ions and nucleic acids. We have studied metal-nucleobase interactions, by using NMR spectroscopy in solution. For the accurate understanding, we tried measurement at low temperature below -30 degrees C. To avoid freezing of the solution, we have extensively searched solution conditions which is suitable for measurements at low temperature, and found several possible conditions. One was the condition which contained H2O and cryoprotectant at low pH for guanosine-5' monophosphate--Cd(II), system, the other was the mixed solvent of dimethyl sulfoxide and N,N-dimethylformamide for guanosine--Cd(II) system. PMID- 12903169 TI - Role of 2'-OH on thermodynamic stability and structure of chimeric oligonucleotides. AB - Thermodynamic parameters of RNA/DNA chimeric duplexes were obtained systematically. Stability of the duplexes including chimeric junctions was context dependent, although CD spectra showed less structural perturbation by the junctions. Nearest-neighbor delta G degree 37 values for these chimeric RNA-DNA junctions suggested that 2'-OH on the 5'-side nucleotide at the junction may be the determinant for the nearest-neighbor stability of the chimeric junction. PMID- 12903170 TI - Theoretical study of substitution effect of the hydrogen bond stability of 9 methylguanine derivatives and 1-methylcytosine. AB - The substitution effect on hydrogen bond stability of the Watson-Crick type base pair between 1-methylcytosine (C) and substitution-introduced 9-methylguanine derivatives (Gx) was studied by an ab initio molecular orbital theory. Introduction of an electron-withdrawing group on the 8-position or on the exo cyclic amino moiety enforced the base pair stability. PMID- 12903172 TI - Interaction of several Ni(III) complexes of peptides with DNA; analysis by DNA fiber EPR. AB - EPR spectra of Ni(III) complexes of GGH, GHG, and GHK were obtained by in-situ oxidation of the Ni(II) complexes in DNA-pellet and on DNA-fibers. A species with an apical coordination of nitrogenous base of DNA was detected for Ni(III) GGH. Both GHG and GHK complexes showed the EPR signals of Ni(III) species when the ligand to metal ion ratio was 2:1. The complexes bind as Ni(III)(N6), NI(III)(N5), and Ni(III)(N4) species on DNA. PMID- 12903171 TI - The interaction of 2-phenylbenzimidazole compounds with DNA: the influence of terminal substituents. AB - We have studied the influence of terminal substituents, amidinium group and N methylpiperazine ring, of 2-phenylbenzimidazole compounds (1-6; Figure 1) on their DNA-binding modes. Experimental results reveal that 1-3 are accepted in intercalation pockets owing to structural flexibility of the N-methylpiperazine ring, while 4-6 bind to the groove of DNA because of steric hindrance between the amidinium group and the DNA phosphate-sugar backbone. PMID- 12903173 TI - RNA motif database catalogued by kink parameters. AB - In the large RNA molecules, the helical stems close to each other with specific arrangement. These specific interactions are mostly identified by the similar mode and can be classified to the several patterns. Several motifs are well characterized at an atomic resolution. The database, structural information about the assemblage and the space distribution of base moieties, can be a useful tool. Kink parameter affords the structural description between the adjacent groups in any irregular geometry. The relative rotational and positional features may be a preferable information to cite the curved or bend phenomenon in not only the successive strand but also the inter strands. The geometrical parameters about hydrogen bond and base stacking would provide the tolerant aspect for multiple strand packing. The tentative database including these geometrical parameters has been constructed. The user can obtain the selected list with the several descriptors for the structure definition and sequence properties. In addition to this nucleic acid system, RNA-protein system would be included. PMID- 12903174 TI - Evaluation of the improvement of IGCR technique. AB - We have improved the protocol of In-Gel Competitive Reassociation (IGCR) technique, one of genome subtraction methods, and developed the apparatus for this technique. The protocol obtained by the fluorescence monitor that we had reported on this symposium last year was added to the improved IGCR technique, which made IGCR method simple and less time consuming for handling. The comprehensive subtractions with genes from twins, one of whom with rheumatoid arthritis, were employed and IGCR libraries were constructed. The analysis of the library clones will be reported. PMID- 12903175 TI - Nonnatural mutagenesis in E. coli and rabbit reticulocyte lysates by using four base codons. AB - Various four-base codons had been developed for introduction of nonnatural amino acids into proteins in an E. coli in vitro translation system. Here, the four base codon mediated incorporation of nonnatural amino acids was investigated in a rabbit reticulocyte lysate. Mutated streptavidin mRNAs containing four-base codons were prepared and expressed in a rabbit reticulocyte lysate in the presence of tRNAs containing corresponding four-base anticodons. Western blot analysis indicates that various four-base codons are efficiently decoded by the corresponding tRNAs in the rabbit in vitro translation system. PMID- 12903176 TI - A novel fluorescent nonnatural amino acid that can be incorporated into a specific position of streptavidin. AB - A novel highly fluorescent nonnatural amino acid (beta-(N-methylanthraniloyl)-L alpha,beta-diaminopropionic acid; nmaDap) was incorporated at the 120th position of streptavidin. The position was directed by a CGGG/CCCG four-base codon/anticodon pair. The incorporation efficiency and the biotin-binding ability of the nmaDap mutant were compared to those of beta-anthraniloyl-L-alpha,beta diaminopropionic acid (atnDap). PMID- 12903177 TI - A new strategy to determine the target DNA sequence of a short peptide. AB - The design of molecules that target desired DNA sequences has been one of the major challenges in the field of molecular recognition. We report here a general strategy for defining the sequence-preference of DNA-binding short peptide by using its heterodimer. Our method successfully identified specific sequences of short peptides derived from native DNA-binding proteins. The usefulness of this approach has been demonstrated by identifying preferred DNA targets for a peptide composed only of D-amino acids. PMID- 12903178 TI - In vitro selection of N peptide-binding RNA on a quartz-crystal microbalance. AB - We report here an in vitro selection of a hairpin loop-RNA bound to N Peptide from bacteriophage lambda on a 27 MHz quartz-crystal microbalance (QCM) to study an interaction between an RNA structure and an RNA-binding peptide. The N Peptide was immobilized on a QCM electrode and specific RNAs binding to the peptide were selected from a GNRNA pentaloop-randomized RNA library with in situ monitoring of selection processes using a QCM. After the 5th round selection, the consensus sequences including a GCGCA loop were obtained from the pentaloop library. This RNA sequence was different from a binding site of a native N Protein (boxB RNA). PMID- 12903179 TI - Roles of histone acetylation in the Dnmt1 gene expression. AB - The DNA methylation plays a key role in the regulation of gene expression, genomic imprinting and X chromosome inactivation and has been shown to be essential for mammalian development. The Dnmt1 is one of the DNA methyltransferases that catalyzed DNA methylation on CpG dinucleotides. The Dnmt1 is constitutively expressed and is required for the maintenance of global methylation after DNA replication. In this study, we investigated the effects of histone deacetylase (HDAC) inhibitor and DNA demethylation agent on promoter activity of mouse Dnmt1 gene in somatic cells. The promoter activity of Dnmt1 gene was increased approximately 2-fold in the treatment of cells by Trichostatin A (TSA) at 1 x 10(-8) M, as compared with that without of treatment of TSA. By contrast, treatment with 5-azacytidine (5aza-C) did not affect the promoter activity of the Dnmt1 gene. These results indicate that the Dnmt1 gene is possibly to regulated by histone acetylation. PMID- 12903180 TI - Real-time detection of PCR products for comparative analysis of expressed genes using module-shuffling TaqMan probes (MTPs). AB - A method for comparative analysis of gene expression was developed. It is based on competitive PCR amplification and real-time detection of PCR products and it uses module-shuffling sequences as "universal TaqMan probes". Namely, cDNA-tagged module-shuffling sequences, which derived from different sources, were amplified in one reaction tube by the same primer set. Two kinds of fluorescent TaqMan probes with different module-shuffling sequences (MTPs) detect their own targets. The method can detect different amounts of expressed genes derived from different sources; accordingly, it was successfully used for comparative analysis of expressed mouse genes. PMID- 12903181 TI - Synthesis of adamantyl naphthalene diimide and its interaction with double stranded DNA. AB - An adamantyl naphthalene diimide derivative (AND) was synthesized as a highly selective double stranded DNA binding reagent. The binding studies with sonicated calf thymus DNA revealed that AND can bind to double stranded DNA by the threading mode, where the two adamantyl moieties are located in the major and minor grooves of DNA duplex separately. In the presence of beta-cyclodextrin (beta-CD), the complex of AND with DNA duplex was stabilized by capping of the adamantyl moieties of AND by beta-CD. In other words, beta-CD serves as a stabilizer for the complex of AND with DNA duplex to result in an enhanced selectivity of AND for double stranded DNA over single stranded DNA. PMID- 12903182 TI - Modified DNA aptamers against sweet agent aspartame. AB - We obtained a modified DNA aptamer against sweetener, aspartame, by in vitro selection method. The modified DNA was prepared from dATP, dGTP, dCTP and a modified dTTP bearing a terminal amino group at C-5 position in place of thymidine by PCR using a hyper thermophilic DNA polymerase, KOD Dash DNA polymerase. The synthetic 102-mer DNA with a 60-mer random region was used as an initial template for the PCR. The PCR-amplified modified DNA library was applied to an aspartame-agarose column, and then the bound modified DNA was eluted from the column for the affinity chromatography selection. Repeating the procedure, we selected the modified DNA aptamer against aspartame. PMID- 12903183 TI - Chemical probing and electron microscopic analysis of alternative DNA structure formed in an inverted repeat sequence. PMID- 12903184 TI - Enzymatic incorporation of an unnatural base pair between 4-propynyl-pyrrole-2 carbaldehyde and 9-methyl-imidazo [(4,5)-b]pyridine into nucleic acids. AB - A hydrophobic unnatural nucleoside of 4-propynylpyrrole-2-carbaldehyde (designated as Pa') was synthesized to improve its affinity with a pairing partner, 9-methyl-imidazo[(4,5)-b]pyridine (Q), in enzymatic incorporation. In single-nucleotide insertion experiments using the Klenow fragment, the substrate of Pa' (dPa'TP) was efficiently incorporated opposite Q in the template strand, as compared to the incorporation of pyrrole-2-carbaldehyde (dPaTP), which was previously developed. PMID- 12903185 TI - Telomerase-inhibitory effects of sugar-modified nucleotide analogs. AB - Telomerase is an endogenous reverse transcriptase that uses its internal RNA moiety as a template for the synthesis of telomere repeats, thus maintaining telomere length. To study the susceptibility of telomerase to sugar-modified nucleotide analogs, inhibition by arabinofuranosylguanine 5'-triphosphate (araGTP), 3'-azido-2',3'-dideoxyguanosine 5'-triphosphate (AZdGTP), 2',3'-dideoxy 2'-fluoroarabino-furanosylguanine 5'-triphosphate (FaraGTP), and their thymine counterparts was investigated. Among these compounds, all dGTP analogs showed potent inhibitory activity against human telomerase. Conversely, dTTP analogs showed moderate or weak inhibition. Partially purified telomerase from cherry salmon testis utilized ddGTP and AZdGTP as substrates into the 3'-terminus of DNA. PMID- 12903186 TI - Genomic analysis of mefloquine-resistant Plasmodium falciparum. AB - We studied sequence polymorphism, expression level of pfmdr1 gene and drug sensitivity in mefloquine sensitive-strain as well as mefloquine-resistant clone/24 to understand underlying drug resistance mechanism. Mefloquine-resistant clone/24 exhibited decreased susceptibility to mefloquine, quinine, halofantrine and artemisinin and increased susceptibility to chloroquine. We analyzed sequence of pfmdr1 gene and found mutation in pfmdr1 on clone/24. Moreover, overexpression of mRNA level of pfmdr1 has been observed in mefloquine-resistant clone/24. Our results explain gene mutation and expression level of pfmdr1 may be associated in mefloquine resistance mechanism. PMID- 12903187 TI - Differences in tyrosine tRNA identity between Escherichia coli and archaeon, Aeropyrum pernix K1. AB - Recognition sites of tyrosine tRNA for tyrosyl-tRNA synthetase from Escherichia coli and extreme thermophilic archaeon, Aeropyrum pernix K1 were examined using various in vitro transcripts. With respect to the long variable arm in E. coli tyrosine tRNA, some base pairs in length was required for tyrosylation. None of the recognition sites were found in the acceptor stem, except the discriminator base A73 in E. coli tyrosine tRNA. In case of A. pernix tyrosine tRNA, C1-G72 base pair and discriminator base A73 in the acceptor region as well as anticodon were base specifically involved in tyrosylation by A. pernix tyrosyl-tRNA synthetase. PMID- 12903188 TI - A novel chitosanase from Bacillus sp. strain K17: gene cloning and expression in Escherichia coli. AB - The gene encoding a novel chitosanase from Bacillus sp. strain K17 was cloned and sequenced. The nucleotide sequence of the gene contained an open reading frame corresponded to a protein of 453 amino acids. The deduced amino acid sequence of the K17 chitosanase exhibited the highest homology to those of family 8 glycanases, suggesting that the enzyme belonged to family 8. PMID- 12903189 TI - Identification of Aquifex aeolicus tRNA (m2(2G26) methyltransferase gene. AB - The modifications of N2,N2-dimethylguanine (m2(2)G) are found in tRNAs and rRNAs from eukarya and archaea. In tRNAs, modification at position G26 is generated by tRNA (m2(2)G26) methyltransferase, which is encoded by the corresponding gene, trm1. This enzyme catalyzes the methyl-transfer from S-adenosyl-L-methionine to the semi-conserved residue, G26, via the intermediate modified base, m2G26. Recent genome sequencing project has been reported that the putative trm1 is encoded in the genome of Aquifex aeolicus, a hyper-thermophilic eubacterium as only one exception among eubacteria. In order to confirm whether this bacterial trm1 gene product is a real tRNA (m2(2)G26) methyltransferase or not, we expressed this protein by wheat germ in vitro cell-free translation system. Our biochemical analysis clearly showed that this gene product possessed tRNA (m2(2)G26) methyltransferase activity. PMID- 12903191 TI - Trans-kingdom conjugation in vitro by using toluenized cells. AB - To establish in vitro trans-kingdom conjugation, we developed a novel direct cell PCR by using toluenized cells which permeate enzymes, primers and substrates. The cells was shown to be successful recipients in conjugation. PMID- 12903190 TI - Effects of triplet repeat sequences on nucleosome positioning and gene expression in yeast minichromosomes. AB - Triplet repeat sequences that cause human hereditary diseases can form a variety of DNA conformations. Since DNA structures act as determinants of chromatin structure, chromatin may be involved in mechanisms of these diseases. To address this issue, we examined effects of triplet repeat sequences on chromatin structure and gene expression in Saccharomyces cerevisiae. We show here that (1) (CTG)12 promotes nucleosome formation, (2) (CGG)12 disrupts an array of positioned nucleosomes, and (3) (GAA)12 has little effect on nucleosome formation. Also, we show that insertion of (CGG)12 increases gene expression of a UAS-less promoter about 10-fold, while (CTG)12 and (GAA)12 have no effect. Thus, expansion of triplet repeat sequences may cause improper expression of disease related genes, through their effects on chromatin structure. PMID- 12903192 TI - Detection of single nucleotide polymorphisms by the combination of nuclease S1 and PNA. AB - A very simple and practical genotyping method has been developed by combining peptide nucleic acid (PNA) and nuclease S1. The portion in the sample DNA, which is complementary with PNA (either completely or partially), is protected from the enzymatic digestion, providing the DNA fragments of predetermined size. The MALDI TOF MASS on them concretely substantiates whether the SNP exists in the DNA of patient or not. By employing appropriate reaction conditions and using a dye as probe, the SNP diagnosis is visually accomplished by naked eyes. PMID- 12903193 TI - Analysis of single nucleotide polymorphisms in uridine/cytidine kinase gene encoding metabolic enzyme of 3'-ethynylcytidine. AB - We investigated single nucleotide polymorphisms (SNPs) in uck2 gene encoding metabolic enzyme of 3'-ethynylcytidine (ECyd) which were associated with drug response of ECyd, and the newly synthesized antitumor ribonucleoside analog. We analized that on exon-intron junction and exon region to affect the qualitative alteration of gene product directly in ECyd sensitive and resistant human cancer cell lines. As the results, cSNP and sSNP were detected in exon 4. In the promoter region, 3 SNPs were detected. Our data seem to be able to give an important knowledge, when ECyd is applied clinically. PMID- 12903194 TI - Detection of NO-induced DNA lesions by the modified aldehyde reactive probe (ARP) assay. AB - When DNA is exposed to NO or HNO2, oxanine (Oxa) is formed as a major guanine lesion. For highly sensitive detection of Oxa using ARP, a probe molecule for DNA damage detection, the reactivity of ARP to Oxa was examined. Oxa site specifically embedded in an oligonucleotide reacted with ARP but it took relatively long time (ca. 24 h) to completely convert Oxa to an ARP-labeled form. PMID- 12903195 TI - Anticancer mechanisms of 1-(3-C-ethynyl-beta-D-ribo-pentofuranosyl) cytosine (ECyd, TAS-106). AB - We investigated the molecular mechanisms of cell death induced by 1-(3-C-ethynyl beta-D-ribo-pentofuranosyl)cytosine (ECyd, TAS-106), a potent inhibitor of RNA synthesis, using mouse mammary tumor FM3A cells and human fibrosarcoma HT1080 cells. ECyd induced the characteristics of apoptosis on these cells, such as morphological changes, DNA fragmentations and caspase-3-like protease activation. General caspases inhibitor, Z-Asp-CH2-DCB inhibited cell death. Interestingly, we also found that ECyd induced rRNA fragmentation. The cleavage pattern of rRNA resembled in that mediated by RNase L. On the other hands, it was suggested that caspase-1, 3, 8 and 9 concerned with ECyd-induced apoptosis through mitochondria. ECyd-induced rRNA fragmentation was inhibited by general caspases inhibitor (Z Asp-CH2-DCB) and caspase-5 inhibitor (Z-WEHD-fmk). So it is clear that caspase-5 (ICErel III/TY), member of ICE (Interleukin-1 beta-converting enzyme) protease, activated pathway concerned with ECyd-induced rRNA fragmentation. These results indicate that antitumor mechanisms of ECyd are involved in caspase-dependent activation of RNase L. rRNA fragmentation may occur one of the death events, as a result of inhibition of RNA synthesis and play an important role in the antitumor activity of ECyd. PMID- 12903196 TI - Mono(ADP-ribosyl)ation of DNA by apoptosis-inducing protein, pierisin. AB - The cabbage butterfly contains a potent cytotoxic protein, pierisin-1, and this protein is suggested to be an ADP-ribosylating toxin, Pierisin-1 effectively transferred an ADP-ribosyl group to DNA, but not to protein, as is the case with other bacteria-derived ADP-ribosylating toxins. Several spectral analyses and independent syntheses indicated that the acceptor site for ADP-ribosylation is N 2 of guanine base. Pierisin-1 induced apoptosis in mammalian cells accompanied by a release of cytochrome c and activation of a variety of caspases, and this apoptosis was inhibited by overexpression of Bcl-2. Pierisin-1 would be a novel DNA-damaging protein. PMID- 12903197 TI - Mechanisms of cell death induced by 5-fluoro-2'-deoxyuridine (FUdR)--necrosis or apoptosis after treated with FUdR. AB - 5-Fluoro-2'-deoxyuridine (FUdR) inhibits thymidylate synthase (TS). The inhibition of TS causes an imbalance of intracellular deoxyribonucleoside triphosphate (dNTP) pools which subsequently induced cell death. We investigated the molecular mechanisms of cell death after treated with FUdR in F28-7 strain (which induced necrosis) and F28-7-A strain (mutant of F28-7 strain which induced apoptosis). After treated with FUdR, we observed different size of DNA fragmentations. F28-7 strain induced DNA cleavaged into 100-200 kbp fragments and F28-7-A strain induced DNA cleavaged into oligonucleosomal sized fragments. In F28-7 strain, FUdR induced the increased mRNA level of c-jun, c-fos and c-myc genes, caspase-3 like protease activity and the changes of mitochondrial membrane potential which were greater and earlier than those of F28-7-A strain. On the other hand, F28-7-A strain induced release of cytochrome c from mitochondrial, but not F28-7 strain. Furthermore, caspase-5 inhibitor was strongly inhibited the cell death of F28-7 strain. We suggest that it is concerned with intensity of intracellular signals in the cell death of F28-7 strain and F28-7-A strain. PMID- 12903198 TI - Identification of AP-2-regulated genes by macroarray profiling of gene expression in human A375P melanoma. AB - Transcription factor AP-2 is a negative regulator of metastasis. Its expression is down regulated with progression of melanoma cells to metastasis. In this study, we performed macroarray profiling of gene expression of human A375P melanoma cells and their derivatives with overexpression of AP-2 and dominant negative AP-2. Such comprehensive analysis lead to an identification of genes such as MMP-2, E-cadherin, melanoma adhesion molecule, early growth response 1, fibroblast growth factor 3, ubiquitin C, histone deacetylase 3 and integrin alpha 5,7, beta 3,5 as regulated by AP-2. Whereas some of these are known as AP-2 regulated genes, the others are not so far. Thus the study reports for the first time identification of new genes regulated by AP-2 that may be involved in metastasis of melanoma. PMID- 12903199 TI - Extensive methylation of CpG island of CYP24 gene in osteoblastic ROS17/2.8 cells. AB - CYP24 is a target gene of 1,25-dihydroxyvitamin D3 (1,25-(OH)2D3) and is induced in the cells expressing vitamin D receptor (VDR) in response to 1,25-(OH)2D3. The osteoblastic ROS17/2.8 cells abundantly express VDR and have been used for the promoter analysis of many of vitamin D3 target genes. However, unlike other cells, ROS cells did not induce CYP24 expression upon 1,25-(OH)2D3 treatment. It has been reported that the methylation of CpG islands of a promoter region is involved in gene silencing. Methylation analysis of the CYP24 gene revealed that the CpG island in 5' part of the transcription unit is extensively methylated. This result suggests that unresponsiveness of the CYP24 gene to 1,25-(OH)2D3 would result from the methylation of the promoter region. PMID- 12903200 TI - Induction of apoptosis in HeLa cells with siRNA expression vector targeted against bcl-2. AB - RNA interference (RNAi) is the process by which double-stranded RNA (dsRNA) directs sequence-specific gene silencing in animal and plant cells. In mammalian cells, 21- or 22-nucleotide (nt) RNAs with 2-nt 3' overhangs small inhibitory RNAs (siRNAs) exhibit an RNAi effect. Very recently, we and others have developed a vector-based siRNA expression system that can induce RNAi in mammalian cells. In this report, to apply this system to oncogene therapy, we tried to suppress the expression of the bcl-2 gene, which is known as a key molecule in the regulation of apoptosis or programmed cell death, by using the siRNA expression system. Western blotting analysis revealed that this siRNA expression vector against bcl-2 suppressed the expression of the bcl-2 protein. Furthermore, HeLa cells which were transiently transfected with the siRNA expression vector against bcl-2 and were subsequently treated with doxorubicin efficiently underwent apoptosis, concomitant with the repression of the bcl-2 gene. These results demonstrate that the siRNA expression vector against bcl-2 has a potential as therapeutic agent for a variety of cancers by down-regulating bcl-2 gene expression. PMID- 12903201 TI - Kinetic analysis of nucleotides incorporated opposite oxidized thymine bases on template DNA. AB - Incorporation of nucleotides opposite the oxidized thymine bases on the template by eukaryotic DNA polymerases alpha and gamma was studied using the synthetic oligonucleotides containing 5-formyluracil or 5-hydroxymethyluracil. Primer extension assay revealed that both polymerases could bypass the oxidized thymine lesion, and incorporated dATP and dGTP opposite 5-formyluracil on the template. On the other hand, dATP was an only nucleotide that was incorporated opposite 5 hydroxymethyluracil by both polymerases as far as examined. These results indicate that 5-formyluracil but not 5-hydroxymethyluracil on template DNA may induce the transition mutation of A:T to G:C in eukaryotes. PMID- 12903202 TI - Application of L-cystein derivative to DNA microarray. AB - S-carboxymethyl-L-cystein derivatives of nucleic acid bases were prepared as DNA chip probe. These compounds in vitro have been found to form stable complex with oligo-DNA and RNA. This paper deals with preparing new DNA chip using L-cystein derivative synthetic nucleotides as probe and immobilized it to quartz plate by photosensitive PVA. Then the chip exposed with FITC labeled target DNA was observed by confocal fluorescence microscope. PMID- 12903203 TI - Regulation of gene expression by sequence-specific alkylating polyamide. AB - In order to investigate the inhibition of gene expression by a new type of hairpin polyamide-CPI conjugate 1, its ability to inhibit transcription in cell free system was investigated. Sequence-selective alkylation of double-stranded DNA by 1 was investigated by denaturing gel electrophoresis using 1000 bp DNA fragment which codes for green fluorescence protein (GFP) under the control of T7 promoter. Analysis of DNA sequence indicated that 1 alkylated predominantly at the site of 5'-AGTCA-3' in coding region of GFP. The transcript by T7 RNA polymerase using the alkylated DNA as a template was analyzed by PAGE. The results clearly indicate that 1 inhibits transcription by alkylation of coding region at a nanomolar concentration. PMID- 12903204 TI - Thermodynamic analysis of hybridization between POPNA and DNA. AB - We have developed a new series of oxy-peptide nucleic acids with pyrrolidine rings of four different types of structural isomers (cis L, cis D, trans L, trans D configurations; POPNAs). Melting curves of adenine nanomers of POPNAs in the presence of the complementary DNA showed the highest Tm value for cis L-POPNA, but in the case of RNA, the highest Tm value was observed for trans L-POPNA. Thermodynamic parameters (delta H and delta S) of POPNA/DNA and POPNA/RNA hybridizations were evaluated from concentration dependence of Tm values. The POPNA/DNA and POPNA/RNA hybridizations were governed by entropy and enthalpy terms, respectively. PMID- 12903205 TI - Formation and stability of alternate-stranded triplex formed with alpha,beta chimeric oligoDNA. AB - Chimeric oligoDNAs composed of alpha-anomeric oligonucleotide and beta-anomeric oligonucleotide with modified nucleobases and/or phosphodiester backbone are synthesized and their ability to form alternate-stranded triple helix with dsDNA was examined. Substitution of normal phosphodiester linkages with phosphorothioate linkages resulted in the prominent destabilization of the alternate-stranded triplex. Meanwhile, the introduction of cationic molecule or intercalative molecule at certain positions of the modified DNA was effective to increase the stability of the triplexes. PMID- 12903206 TI - Acceleration of DNA strand exchange reaction by cationic polymers. AB - In previous reports, we have demonstrated that the cationic comb-type copolymers (PLL-g-Dex) having a poly-L-lysine main chain and dextran side chains stabilized DNA duplex and triplex. Furthermore, the copolymer was found to accelerate more than 20,000 folds strand exchange reaction between 20 bp duplex and its homologous single strand (1). This study was designed to inspect structural factor of polycationic moieties in acceleration effects. To this end, polypeptides containing lysine or arginine moieties as cationic groups were examined in their potency to accelerating strand exchange reaction. It was shown that arginine-rich peptide showed higher accelerating effect than lysine peptide when total positive charges of peptides were kept constant. PMID- 12903208 TI - Functional regulation of biomolecule using DNA-conjugation. AB - 3'-methacryloyl-oligodeoxynucleotide having antisense sequence against messenger RNA of green fluorescent protein and N-isopropylacrylamide were co-polymerized. The conjugate can reversibly change its conformation at around a transition temperature, similarly to N-isopropylacrylamide homopolymer. We achieved artificially regulation of gene expression depending on temperature using the conjugate. PMID- 12903207 TI - In vitro selection of RNA aptamers that bind to domain II of HCV IRES. AB - The translation of HCV starts at the internal ribosomal entry site (IRES) within the 5' untranslated region and domain II of IRES is essential for translation activity. However, the information of function is limited. We attempted to obtain RNA that bind HCV IRES domain II. Selected aptamers will give us much information, such as RNA-RNA interaction between IRES and other cellular RNAs as well as target sites for anti-HCV drugs. We developed a novel selection method using biotinylated DNA probe. This method can be applied to many RNAs and would identify the most suitable target position experimentally and simply. The binding kinetics of aptamers were analyzed by using Biacore. Furthermore, inhibition of translation by aptamers was analyzed. PMID- 12903209 TI - Regulation of bacterial RNase P ribozyme reaction by divalent cation and guide DNA. AB - The RNA subunit of bacterial ribonuclease P (RNase P) is a ribozyme which can cleave a canonical cloverleaf tRNA precursor and a hairpin RNA with a CCA-3' tag sequence as its substrate. With high concentration of Mg ion, the ribozyme as well as holo enzyme internally cleaves certain tRNAs in vitro. We denoted this unusual reaction as hyperprocessing. By controlling magnesium ion concentration for the reaction and also by forcing the RNA shape with external guide DNAs, we could regulate the hyperprocessing reaction by the bacterial RNase P enzymes. These techniques will lead the RNase P ribozyme to more designable and more applicable RNA-cleaving enzyme. PMID- 12903210 TI - Analyses of kinetic solvent isotope effects of a hammerhead ribozyme reaction in NH4+ and Li+ ions. AB - Hammerhead ribozymes have been considered to be divalent-metalloenzymes. However, this was recently questioned by the finding that the reaction can proceed without any divalent metal ions in the presence of high concentrations of monovalent ions such as NH4+/Li+ ions. Thus, one might think divalent metal ions are not involved in chemical step in the catalytic mechanism. To investigate the involvement of the monovalent ions, we analyzed the deuterium solvent isotope effects in the reactions. Our present analysis indicates a proton transfer(s) occurs only in the NH(4+)-mediated reaction, not in the Li(+)-mediated. Most simple interpretation is that NH4+ works as a general acid catalyst and Li+ as Lewis acid catalyst. This suggests hammerhead ribozymes can change catalyst upon their surrounding conditions. PMID- 12903211 TI - A functional gene discovery in cell differentiation by hybrid ribozyme and siRNA libraries. AB - Recently, we developed a gene discovery system that can identify functional genes using a randomized hybrid ribozyme library. In this system, inhibition of the expression of a particular gene by active ribozymes was reflected by a change in a particular phenotype, the method allowed the identification of functional genes. In the case of identification of functional genes for apoptosis pathways, we identified many pro-apoptotic genes in TNF-alpha and Fas-mediated apoptosis pathways. In this study, we tried to identify the functional genes that are necessary for the retinoic acid (RA)-induced cell differentiation using randomized ribozyme and siRNA libraries. We succeeded to identify the several differentiation factors. Therefore, our gene discovery system based on randomized ribozyme and siRNA libraries are high potential to identify the differentiation and undifferentiation factors in the post genome era. PMID- 12903212 TI - Synthesis of a novel anticancer prodrug designed to target telomerase sequence. AB - A curcumin conjugate viz. 1,7-bis (4-O-glycinoyl-3-methoxyphenyl)-1,6- heptadiene 3, 5, dione (I) was synthesised and after attachment of linker (-CH2-CH2-OH) phosphitylated. This unit was attached to the deoxy-11 mer, 5'-GTT AGG GTT AG-3', a complementary sequence of telomerase RNA template. Its hybridisation with the telomerase RNA sequence, 5'-CUA ACC CUA AC-3' has been studied employing Tm. PMID- 12903213 TI - A new approach for the cooperative chemical evolution of nucleic acids and proteins under the primitive earth environment. AB - In order to evaluate cooperative chemical evolution of nucleic acids and proteins, we have examined possible activity of thermal copolymers of amino acids (TCAA) for prebiotic formation and degradation of RNA. Several types of TCAA involving aromatic amino acids were prepared and the influence on the template directed formation of oligoguanylate and prebiotic ribonuclease activity were kinetically studied. PMID- 12903214 TI - Selection of RNA-binding peptides containing Arg-rich motif. AB - In order to search the RNA-binding peptides, selection from randomized peptides containing Arg-rich motif was carried out using rRNA-immobilized column. As a result of the selection for 16S- and 23S-rRNA from E. coli, six peptides were identified to be the rRNA-binding peptide by MALDI-TOF MS. These peptides contained R, N, Q, or G at three randomized positions. This result indicates that amino acids with a positive residue or an amide residue might be essential for peptides to recognize the RNA. PMID- 12903215 TI - DNA topology: topological transformation of DNA by reaction of cisplatin-DNA-core histone complexes with human DNA topoisomerase. AB - Cis-diamminedichloroplatinium(II), an active antitumor agent binds to core histone -SV40 DNA complexes prepared by reaction of DNA with core-histone, and alters the fiber-like structure into the loosened structure. In order to model the cisplatin-modified chromatin complexes in cell, the complexes were reacted with human DNA topo II. We found the generations of unique topologically isomers such as trefoil knot (and catenane) and pseudo catenane (and pseudo knot) by reaction of cis-DDP--linearDNA--core-histone complexes with DNA topo I. The results are discussed in relation with a possible recombinational role of topo II (or topo I) on the reaction with cis-DDP--DNA--core-histone complexes. PMID- 12903216 TI - A novel heme-DNA coordination complex and its stability. AB - Heme, iron(III)-protoporphyrin IX complex, and a parallel-quadruplexed d(TTAGGG) have been shown for the first time to form a coordination complex with a guanine bound to heme iron as an axial ligand. PMID- 12903217 TI - Gene mutation assay using a MutS protein-modified electrode. AB - A novel electrochemical biosensor for gene mutation detection was developed using a DNA mismatch recognizing protein MutS from E. coli. The MutS protein was immobilized onto an Au electrode by coordination of His-tag at its C-terminus to vacant sites of Ni(II)-nitrilotriacetato complex attached to the surface of electrode. When a target DNA duplex having a mismatch site was captured by the MutS protein on the electrode, the electrostatic repulsion arose between polyanionic DNA duplexes and negatively-charged ferrocyanide/ferricyanide redox couple ions. Consequently, their redox peak currents on a cyclic voltammogram with the Au electrode drastically decreased depending on the concentration of the target DNA according to the redox couple-mediated artificial ion-channel principle. Using this assay, we could detect GT mismatch and deletion mutation in the double-stranded DNA. PMID- 12903218 TI - Useful technique for DNA-stretching and fixation. AB - We report on a useful technique for reproducibly straightening and fixing DNA molecules. When a droplet of DNA solution on surfaces is sucked up by pipet, surface tension at the moving air-water interface is sufficient to stretch the molecules. The point of this technique is the interaction control between surfaces and DNA molecules by polymer coating. With polymers containing pi conjugation units (polyphenazasiline, PPhenaz and poly(vinylcarbazole), PVCz), many DNA were nicely stretched and fixed on surfaces. Furthermore, structural changes of pi-units in polymers affected DNA-stretching and fixation. By using atomic force microscopy (AFM) and scanning near-field optical/atomic force microscopy (SNOM/AFM), the feasibility of this technique for DNA observations was demonstrated. PMID- 12903219 TI - An anthracene derivative carrying ferrocenyl moieties at its 9 and 10 positions as a new electrochemically active threading intercalator. AB - A ferrocenyl anthracene derivative (FAN) was synthesized by connecting ferrocene moieties to the amino termini of its substitutents at 9 and 10 positions, aiming at an improved hybridization indicator. Spectrophotometric experiments revealed that FAN can bind to double stranded DNA (dsDNA) by threading intercalation. Evaluation of FAN in the dsDNA preference as hybridization indicator revealed that FAN has higher preference for dsDNA than ferrocenyl naphthalene diimdie (FND) reported previously. PMID- 12903220 TI - Structural change of G-quartet by 5'-elongation of telomere DNA oligomer. AB - Structural change of G-quartet formed by telomere G-rich DNA oligomers are investigated by NMR and CD spectroscopy. G-quartet structure changes depending on the length of 5'-terminal sequence. PMID- 12903221 TI - Development of HIV-1 dependent gene expression vector with the Cre/loxP (ON) system. AB - We used the HIV-1 5'LTR and the Cre/loxP system to develop an anti-gene expression system. The LTR promoter of HIV-1 has a specific activity that includes the intermediary region of gag, as shown in a previous report. We constructed the U5-region of HIV-1 as the target of the ribozyme expression vector (pCre/loxP-Rz), with the Cre/loxP system under the control of this LTR promoter. The function of this vector is to induce the HIV-1 dependent ribozyme mediated inhibition in a dose-responsive manner. Furthermore, ribozyme mRNA expression was detected in the presence of pNL4-3 in HeLa-CD4+ cells. These studies are expected to yield novel applications of antiviral strategies for HIV 1. PMID- 12903222 TI - RNA cleavage by a mammalian tRNA 3'processing endoribonuclease (3'tRNase) reduces HIV-1 expression. AB - We examined the suppression of HIV-1 expression by cleavage of the HIV-1 RNA gene, using a mammalian tRNA 3'processing endoribonuclease (3'tRNase) as an External Guide Sequence oligozyme (EGS) in vivo. The enzyme can also recognize and cleave a target RNA that forms a pre-tRNA like complex with a substrate RNA. We designed an EGS to specifically recognize the packaging signal region (p.s.) and the upstream portion of the gag region (g.s.) of HIV-1 mRNA, and constructed EGS expression vectors that used the tRNA(met) promoter as an expression cassette for the EGS. Their cleavage activities were assessed in cell culture, using a transient assay system. The EGS efficiencies were determined by co-transfection of the EGS expression vectors and the HIV-1 gene plasmid vector (pNL-luc) into COS cells. The assay results showed a significant inhibition of the HIV-1 gag p24 antigen expression. In addition, we constructed MLV-based VSV-G pseudotyped retrovirus vectors that express the EGS. They were co-infected into MT-4 cells with the HIV-1 virus (NL4-3). These EGS expression retrovirus vectors exhibited a similar inhibitory effect. PMID- 12903223 TI - Patient safety: lessons from a novice. PMID- 12903224 TI - Assessment and management of pain and distress in the neonate. AB - Pain management is an integral focus of neonatal care. This article reviews the physiology and impact of neonatal pain and distress and pain assessment tools, as well as clinical interventions and current controversies in the management of pain and distress in neonates. Current guidelines to enhance the recognition and treatment of pain are highlighted. PMID- 12903225 TI - Scrotal and inguinal masses in the newborn period. AB - Scrotal or inguinal masses may represent inguinal hernias, hydroceles, or testicular torsion and are common findings in the newborn period. The clinician is challenged to differentiate between normal, abnormal, atypical, and pathologic findings and to seek urgent surgical consultation when compromised bowel, testis, or ovary is suspected. This issue of Focus on the Physical offers a review of the embryologic development of the processus vaginalis and testicular attachments to enhance the clinician's understanding of the development of these conditions. Systematic advanced physical assessment techniques will be provided along with pictures of common clinical findings. A brief discussion of the diagnostic studies that aid in differentiation of scrotal and inguinal masses and the clinical implications of each of these conditions will be discussed. PMID- 12903227 TI - Perspectives on parenting in the NICU. PMID- 12903226 TI - Comparison of two microenvironments and nurse caregiving on thermal stability of ELBW infants. AB - PURPOSE: This study tested a novel bioinstrumented radiant warmer (RW), designed to provide a reduced-intervention nursing caregiving environment (RINCE) on the ambient and body surface temperature stability of extremely low birth-weight (ELBW) infants (< 1,000 g) during caregiver disruptions. The RINCE was compared with a control bed consisting of a standard RW, modified with a Plexiglas hood (Rohm and Haas, Philadelphia, PA) and an external humidity source. SUBJECTS: The sample consisted of 10 extremely low birth-weight infants. The mean gestational age was 25.7 weeks (+/- 1.4 weeks). Birth weight ranged from 570 to 880 g: mean birth weight was 730 g (+/- 95.14 g). The mean Scores for Neonatal Acute Physiology was 21.4 (+/- 6). DESIGN: This clinical study used a small n sample, crossover time-series design. Infants were randomized to treatment order (control bed v RINCE) on the first day of life. A washout period between microenvironments was observed, and then each infant was placed into the other condition. METHODS: Each infant was studied for 5 hours in each treatment condition (control bed v RINCE). Abdominal skin temperatures, heel temperatures, ambient temperatures, and relative humidity (RH) were recorded by a computerized data acquisition system every 60 seconds for hours 12 to 24 of life. Caregiver interventions, microenvironment disruptions, or both were manually coded in the data set. PRINCIPAL RESULTS: The mean number of minutes abdominal temperatures were less than 36.5 degrees C was not significantly different in either microenvironment (P = 0.48). The mean number of minutes that heel temperatures were less than 35.2 degrees C and central-peripheral differences were greater than 2 degrees C was significantly less in the RINCE (P < 0.04 and P < 0.0001, respectively). Although the number and duration of caregiving disruptions (P = 0.71), number of procedures (P = 0.30), and the duration of interventions (P = 0.56) were not significantly different in either environment, the ambient air temperature (P < 0.001) and RH (P < 0.001) were significantly less variable in the RINCE during and after caregiving disruptions. CONCLUSIONS: This study identified the extreme inter-dependence of the ELBW infant's temperature and the ambient environment. The RINCE significantly improved ambient temperature variability, RH variability, and the infant's peripheral and delta temperature (delta T) stability, particularly during and after caregiver disruptions. PMID- 12903228 TI - A parent's guide for advocacy and involvement. PMID- 12903229 TI - Beyond the black cloud. PMID- 12903230 TI - Twenty-first century New Year's resolutions for health care. PMID- 12903231 TI - Evidence-based feeding guidelines for very low-birth-weight infants. AB - Clinical practice guidelines (CPG) for the nutritional management of premature infants are limited. This project focused on the development of a research-based enteral feeding CPG for infants of < 1,500 g. The CPG was based on an extensive literature review and developed through a process of consensus decision making by a team of clinical researchers. Infants that weigh < 1,000 g initiate minimal enteral nutrition (MEN) at 48 hours; nutritional feedings begin on day 5 to 6 of life. For infants between 1,000 and 1,500 g, nutritional feedings begin at 48 hours and are advanced at a rate of less than 30 mL/kg per day. The benefits and risks of continuous versus intermittent nasogastric tube feeding were inconclusive; therefore, the CPG does not stipulate a feeding method. Breast milk is used preferentially, and specific guidelines for the definition and management of feeding intolerance are provided. A follow-up study testing this CPG has been completed and is published in the original research section of this issue. PMID- 12903232 TI - Look before you clamp: delivery room examination of the umbilical cord. AB - The umbilical cord is a critical connection between the embryo (and later, the fetus) and the placenta. The umbilical cord houses the blood vessels that are responsible for nourishing the fetus. Proper umbilical cord function is essential for growth and development before birth. A review of the embryologic origin of the umbilical cord aids in understanding abdominal wall and umbilical cord defects that can present in the newborn period. In the delivery room, inspection of the umbilical cord is an integral part of the first minutes of life. Any abnormality either within the cord structure or in the areas surrounding the base of the cord may necessitate a delay in shortening the cord. Surgical consultation may also be indicated. This issue of Focus on the Physical will provide a step-by step guide to cord assessment in the delivery room setting. Pictures showing normal, atypical, and abnormal umbilical cords and common abdominal wall defects will be presented along with a brief discussion of the significance and clinical implications of each of these findings. PMID- 12903233 TI - A matched cohort study of feeding practice guidelines for infants weighing less than 1,500 g. AB - PURPOSE: To evaluate the efficacy and safety of clinical practice guidelines (CPG) for the nutritional management of infants weighing < 1,500 g. SUBJECTS: Infants weighing < 1,500 g (n = 200) admitted to the NICU who had no major congenital anomalies were enrolled. DESIGN AND METHODS: A before-and-after matched cohort study was conducted during 1996/1997 and 1998/1999 enrolling infants in a Standard Practice (SP) group and CPG group, respectively. Weight stratified CPG were introduced between these 2 study periods. Data on the first 100 babies who could be matched for birth weight and gestational age were analyzed. Data collection continued until full feedings were established and tolerated for 48 hours or the infant was discharged from the hospital, whichever came first. PRIMARY OUTCOME MEASURE: Of the 200 infants in the study (median gestational age 28 weeks), 142 infants attained full feedings. The median time to full feedings was 15 days in both groups, and a paired sample t test showed no significant difference between the 2 groups (P = 0.35). PRINCIPAL RESULTS: No statistically significant differences in the age of feeding commencement, number of feeding interruptions, days on total parenteral nutrition, days to regain birth weight, age at discharge, incidence of sepsis, necrotizing enterocolitis, or use of erythromycin were found. CONCLUSIONS: The CPG was a safe alternative to standardize nutritional practices in the NICU. The lack of differences between groups shown in this study is likely related to gut immaturity limiting the infant's response to changes in feeding practices, inconsistent use of the guidelines, confounding factors, the small sample size, or the similarity between SP and the CPG. PMID- 12903234 TI - Immunization essentials: special considerations for premature infants and newborns. PMID- 12903235 TI - A teaching guide for immunizations. PMID- 12903236 TI - Getting back to sleep. PMID- 12903237 TI - Communicating patient information in the HIPAA era: the good news and the bad news. PMID- 12903238 TI - Where did all the fun go? PMID- 12903239 TI - The gene encoding a novel halorhodopsin-like protein of extremely halophilic archaeon Haloarcula japonica strain TR-1. AB - The gene encoding the halorhodopsin (hR)-like protein was cloned and sequenced from Haloarcula japonica strain TR-1. The structural gene consisted of an open reading frame of 828 nucleotides encoding 276 amino acids. The deduced amino acid sequence of the Ha. japonica hR-like protein showed the highest homology to those of cruxhalorhodopsins. Furthermore, we have demonstrated that the Ha. japonica chR gene is transcriptionally induced by light. PMID- 12903240 TI - Mutations induced by glyoxal and methylglyoxal in mammalian cells. AB - To investigate the mutation spectra of glyoxal and methylglyoxal in mammalian cells, we analyzed mutations in a bacterial suppressor tRNA (supF) gene in the shuttle vector plasmid pMY189. The cytotoxicity and the mutation frequency increased according to the doses of glyoxal and methylglyoxal. The majority of glyoxal-induced mutations (65%) were base-pair substitutions, in which G:C-->C:G transversions were predominant. In the mutants induced by methylglyoxal, multi base deletions were predominant (50%), followed by base-pair substitutions (35%), in which G:C-->C:G and G:C-->T:A transversions were predominant. PMID- 12903241 TI - Analysis of histidine-dependent antitermination in Bacillus subtilis hut operon. AB - We have previously shown that a positive regulator, HutP, of Bacillus subtilis hut operon is a RNA binding protein. Here, we report precise binding site of HutP in cis-regulatory region on hut mRNA and the role of HutP in histidine-dependent antitermination of hut expression. Ethylnitrosourea modification interference assay showed that four binding sites of HutP were found in the cis-regulatory sequences and were located at the stem and the internal loop of an antiterminator structure. In vitro transcription assay indicated that HutP suppressed transcription termination in the presence of histidine. These results suggest that HutP function as an antiterminator in response to the presence of histidine. PMID- 12903242 TI - Escherichia coli proline tRNA: structure and recognition sites for prolyl-tRNA synthetase. AB - A major proline tRNA was purified from bulk Escherichia coli A19 tRNA by affinity chromatography with a biotinylated DNA probe. Its nucleotide sequence including modified nucleotides was determined by the post-labelling technique. In order to study the recognition sites of this proline tRNA for prolyl-tRNA synthetase, various mutant transcripts were prepared using an in vitro transcription system with T7 RNA polymerase. Based on the results of in vitro kinetic analyses of mutant transcripts, it was concluded that the second and third letters, G35 and G36, of the anticodon, G37 of the anticodon loop, the discriminator base A73, G72 of the acceptor stem, G49 and U17A that existed in the corner of an L-shaped structure are the recognition sites of proline tRNA for prolyl-tRNA synthetase. PMID- 12903243 TI - Construction of an efficient expression vector for coupled transcription/translation in a wheat germ cell-free system. AB - Using the expression vector, pEU, which we have constructed, highly efficient in vitro protein synthesis can be achieved: The system works for 150 hours and without further template addition once the reaction has started, yielding 5 mg of enzymatically active protein in a 1 ml reaction. PMID- 12903244 TI - Enzymatic properties of Escherichia coli and human 7,8-dihydro-8-oxoguanine DNA glycosylases. AB - Oxidative damage to DNA generates aberrant guanine bases such as 2,6-diamino-4 hydroxy-formamido-pyrimidine (Fapy) and 7,8-dihydro-8-oxoguanine (8-oxoG). Although synthetic oligonucleotides containing a single 8-oxoG have been widely used to study enzymatic processing of this lesion, the synthesis of oligonucleotides containing Fapy as a unique lesion has not been achieved to date. In this study, an oligonucleotide containing a single 2,6-diamino-4-hydroxy 5-(N-methyl)formamido-pyrimidine (me-Fapy, a methylated derivative of Fapy) was prepared by a DNA polymerase reaction and the subsequent alkali treatment. The repair activity of Fpg and hOGG1 proteins were compared using oligonucleotide substrates containing me-Fapy and 8-oxoG. PMID- 12903245 TI - DNA binding of a basic leucine-zipper protein with novel folding domain. AB - DNA-binding proteins frequently utilize short alpha-helices as their critical DNA recognition elements. In this research, we have employed the structure-based design to construct a small domain that could target the specific DNA sequences recognized by the yeast transcriptional activator GCN4. The new DNA binding motif recognizes specific DNA sequences as a dimer with high affinity and specificity under the physiological conditions. PMID- 12903246 TI - New thermodynamic characterization and transition mechanism of DNA duplex formation. AB - A new transition mechanism of DNA duplex association was proposed and a segregated transition model (STM) was further derived. The experimental results in various molar ratios showed that the duplex association transition is imperfect and the thermodynamic properties and self-transition behavior of single strands exert a significant influence on DNA duplex formation. PMID- 12903247 TI - Synthesis and structure-activity relationships of selective ligands for P3 purinoceptor-like protein (P3LP). AB - We examined the structure-activity relationship of various 5'-N-substituted carboxamidoadenosine derivatives toward P3 purinoceptor-like protein (P3LP), which has affinity for both adenine nucleosides and nucleotides. We discovered a hydrophobic binding region near the 5'-N-substituted-carboxamide group. From the linear alkyl N-substituted derivatives, 1-(adenin-9-yl)-1-deoxy-N-n-pentyl-beta-D ribofuranuronamide (6) was found to be the most potent ligand. In the series of the N-cycloalkyl derivatives, 1-(adenin-9-yl)-1-deoxy-N-cyclohexyl-beta-D ribofuranuronamide (8) was the strongest ligand. We also examined the receptor selectivity for the selected nucleosides 6 and 8 with 1 (HAK2701) and N ethylcarboxamidoadenosine (NECA) versus P1 purinoceptor subtypes, such as adenosine A1, A2A, A2B, and A3 receptors and found 8 is the most selective ligand for P3LP. PMID- 12903249 TI - Synthesis of 2'-O-deuteriomethyl ribonucleosides Gm-d3, Am-d3, Um-d3 and Cm-d3. AB - The synthesis of 2'-O-deuteriomethyl ribonucleosides by iodomethane-d3 (99.5 + atom % D) deuteriomethylation of 3',5'-O-tetra(isopropyldisiloxane)-diyl nucleosides, followed by deprotection, is described. PMID- 12903248 TI - Stereospecific synthesis of alpha-anomeric pyrimidine nucleoside. AB - A facile stereospecific synthetic method for alpha-anomeric 2'-deoxypyrimidine nucleoside unit utilizing aminooxazoline derivative of ribofuranose was investigated. Thus, easily accessible riboaminooxazoline derivative prepared by ribose and cyanamid was allowed to react with ethyl alpha-bromoethylacrylate to give corresponding adduct. The adduct was cyclized by strong base such as potassium t-butokiside. The resulted 2,2'-cyclonucleoside was then treated with acetyl bromide followed by n-butyltin hydride to give alpha-anomeric 3',5'-di-O acetylthymidine. 3',5'-Di-O-acety groups of the nucleoside were easily removed by the action of excess of triethyl amine in methanol. Essentially same procedure afforded corresponding 2'-deoxyuridine, which was further, converted to alpha anomeric 2'-deoxycytidine. PMID- 12903250 TI - Study on highly diastereoselective synthesis of (2'R)-2'-deoxy[2'-2H]guanosine. AB - To develop an efficient method for the synthesis of a highly diasteroselective (2'R)-2'-deoxy[2'-2H]guanosine (1), studies of organic chemical conversion from 2'-bromo-2'-deoxy-N2-Isobutyryl-3',5'-O-TIPDS-guanosine (2) to 1 and a biological transdeoxyribofuranosylation of (2'R > 98% de)-2'-deoxy[2'-2H]uridine (4) were carried out. As the results, a highly diastereoselective synthesis of 1 was achieved by a biological transdeoxyribofuranosylation between 2,6-diaminopurine and 4 by the use of Enterobacter aerogenes AJ-11125, followed by treatment with adenosine deaminase. The results will be described in detail. PMID- 12903251 TI - Synthesis of oligonucleotide derivatives containing a bis-pyrene residue in the main chain. AB - The oligonucleotide having the bis-pyrene residue in the main chain was synthesized. The preparation of the bis-pyrene was started from the conversion of 2,2-bis-(bromomethyl)-1,3-propanediol into the protected bis-amino derivative. The reaction of the bis-amino derivative with 1-pyrenebutyric acid using DCC/HOBT afforded the desired bis-pyrene. This compound was then converted to the protected phosphormidite. The oligonucleotides possessing the bis-pyrene were synthesized by using the amidite. The oligonucleotides having the bis-pyrene residue can bind to DNA sequence in an aqueous solution to give the duplex with comparable thermal stability as that of the unmodified DNA/DNA duplex. The significantly enhanced pyrene-excimer fluorescence was observed upon hybridization of the bis-pyrene modified oligonucleotides with DNA. PMID- 12903252 TI - Synthesis of 2-5As possessing base-modified adenosines and their activities to human recombinant RNase L. AB - The unique 2',5'-oligoadenylate (2-5A) acts as a potent inhibitor of translation in vertebrate cells through the activation of a constituent latent 2-5A-dependent endoribonuclease (RNase L). This 2-5A system plays a major role in the interferon natural defense mechanism against viral infection. We report the syntheses of base-modified adenosine-substituted 2-5A derivatives, their interaction with recombinant human RNase L and their biological stability. PMID- 12903253 TI - Chemical synthesis of hydroxymethylphosphonate alpha-DNA. AB - In this paper, we report the synthesis of hydroxymethylphosphonate alpha-DNAs and related compounds by use of the H-phosphonate method. These modified alpha-DNAs were designed to improve the inherent poor solubility of well-known methylphosphonate alpha-DNAs by introduction of a more hydrophilic hydroxymethylphosphonate function. The hybridization ability of hydroxymethylphosphonate alpha-DNAs was studied. We also report a novel strategy for the synthesis of alpha-thymidine by use of C1'-epimerization. The details of the neighboring effect of various 5'- and 3'-hydroxyl protective groups such as carbamoyl groups on the beta-->alpha conversion will be described. PMID- 12903254 TI - Synthesis of oligonucleotides using the 2-(levulinyloxymethyl)-5-nitrobenzoyl group for the 5'-position of nucleoside 3'-H-phosphonate and -H-phosphonothioate derivatives. AB - Synthetic studies on phosphodiester, phosphorothioate, and phosphorodithioate linked oligonucleotides in terms of 2-(levulinyloxymethyl)-5-nitrobenzoyl (LMNBz) group as the base-labile protecting group for the 5'-hydroxyl groups of nucleoside 3'-H-phosphonate and -H-phosphonothioate derivatives, are described. PMID- 12903255 TI - Control of electron transfer in DNA by peptide nucleic acids (PNA). AB - The one-electron oxidation of PNA-DNA hybrid containing G-triplet sequence was examined. In DNA duplex G-triplet was selectively cleaved by oxidation, whereas in PNA-DNA hybrid cleavage efficiency was extremely lowered. These result suggested that cleavage efficiency of PNA-DNA hybrid was different from that of B form DNA duplex. PMID- 12903256 TI - Convenient synthesis of arabinonucleoside containing oligodeoxyribonucleotides. AB - C2 substituted arabinofuranosyluracil derivatives were synthesized and its incorporations into DNA were easily carried out by using post-synthetic modification. PMID- 12903257 TI - Design and synthesis of the novel cross-linking reagents triggered by the triple helix formation. AB - In our attempt to new nucleobase analogs capable of interstrand cross-linking, we developed 2-amino-6-vinyl purine analog (1). The oligonucleotides incorporating 1 showed efficient interstrand cross-linking with selectivity toward cytidine at a target site. In this paper, we describe the design of the new cross-linking reagents (2) bearing 2-amino-6-vinyl purine motif, and triplex-directed alkylation with 2 to double-stranded DNA. PMID- 12903258 TI - Interaction of peptides derived from RecA with single-stranded oligonucleotides containing 5-formyl-2'-deoxyuridine. AB - We report the first example of chemical cross-linking of 5-formyl-2'-deoxyuridine containing oligonucleotides with oligopeptides through a Schiff base formation. Twenty amino acid residue peptides investigated here were derived from the DNA binding site of RecA protein. We have demonstrated that the lysine residue placed at the 6th or 8th position from the N-terminus of the peptide directly contacts with DNA. PMID- 12903259 TI - Synthesis of 6-(2-thienyl)purine nucleoside derivatives toward the expansion of the genetic code. AB - Unnatural bases specifically pairing with pyridin-2-one, 2-amino-6-(2-thienyl) purine and 2-amino-6-(2-furanyl)purine, were newly designed to replace 2-amino-6 (N,N-dimethylamino)purine. It was expected that these novel purine analogues, as compared with 2-amino-6-(N,N-dimethylamino)purine, might reduce the interference in the stacking interactions with the neighboring bases in a duplex and improve the efficiency of the enzymatic incorporation of the nucleoside triphosphate of pyridin-2-one opposite these unnatural bases. The syntheses of these nucleoside derivatives and the DNA fragments were examined. PMID- 12903261 TI - Synthesis and properties of novel acyclic nucleotides. AB - Acyclic adenosine and thymidine analogs derived from L- and D-threonie were synthesised and incorporated into oligonucleotides by automated protocols using a standard phosphoramidite method. UV melting experiments with thus obtained oligonucleotides showed that incorporation of those acyclic nucleosides did not destabilize the hybrid duplexes and that the stabilities of them are influenced by the stereochemical structures of acyclic analogs. Modification of 3'-end of oligonucleotide with acyclic analogs protected the oligonucleotide against 3' exonuclease. PMID- 12903260 TI - The synthesis of pyrrole(Py)/imidazole(Im) polyamide CPI conjugates which possess DNA interstrand cross-linking activity. AB - Covalent dimers of ImPyLDu86 with various linkers were synthesized and their DNA interstrand cross-linking abilities were examined. Denaturing polyacrylamide gel electrophoresis indicated that dimer molecule with tetramethylene linker efficiently produced interstrand cross-link only in the presence of appropriate partner PyIm triamides. PMID- 12903262 TI - Structure and affinity of DNA binding peptides. AB - Artificial peptides designed to form alpha-helical, beta-turn, antiparallel beta sheet and beta-hairpin structures which are among the motifs most frequently found in natural DNA/RNA binding proteins were synthesized and their characteristic features were examined in the presence or absence of double or triple stranded DNA by means of UV melting experiments, CD spectra, SPR measurements. It was revealed that amphiphilic character arising from the specific secondary structures and positive charge in the hydrophobic face of peptides played an important role in the interaction with DNA, and that hybrid duplex and triplex were intensively stabilized by the cationic amphiphilic peptides. It was also found that these peptides could protect dsDNA against DNase 1 digestion. These results indicate that structurally designed amphiphilic peptides synthesized in the present study can be powerful tools for antisense and antigene strategies. PMID- 12903263 TI - Dynamics of the fluorescence properties of pyrene residues appended to oligonucleotide hybridization probes. AB - The dynamic and static properties of the fluorescence of a pyrene-introduced oligonucleotide 16 mer and its hybrid with a target 32 mer. Their fluorescence quantum yields (< 1%) were much weaker than that of unsubstituted pyrene and their fluorescence lifetime of the major decay components were less than 1 ns. The rapid fluorescence quenching was due to the interaction between the fluorophore and bases in the oligonucleotides. The fluorescence of pyrene was quenched efficiently by TMP and slightly by AMP. The quenching by CMP and GMP were the intermediate case. PMID- 12903264 TI - DNA binding properties of DAPI (4',6-diamidino-2-phenylindole) analogs having an imidazoline ring or a tetrahydropyrimidine ring: groove-binding and intercalation. AB - DAPI analogs containing an imidazoline ring or a tetrahydropyrimidine ring have been synthesized to study DNA binding properties. Spectroscopic (absorption, CD, flow dichroism and fluorescence) and viscosity measurements indicate that DAPI analogs interact with DNA both by intercalation and by groove binding. The solution structures of complexes between DAPI analog and DNA oligomers have been characterized by proton NMR spectroscopy. PMID- 12903265 TI - Analysis of the RNase H activity by fluorescence resonance energy transfer. AB - The RNase H activity of HIV-1 reverse transcriptase was examined using chemically synthesized deoxyribo.ribo-oligonucleotide hybrid duplex labeled with the fluorescence donor at the 5'-end and with the fluorescence acceptor at the 3'-end of DNA strand as a substrate. Fluorescence resonance energy transfer (FRET) between these fluorescent dyes was used to analyze the rate of the enzymatic reaction. Under excitation of the donor dye, that is 6-carboxyfluorescein (6 FAM), at 490 nm, the increase of the fluorescence resulting from the acceptor dye, that is 6-carboxytetramethylrhodamine (TAMRA), at 578 nm, was observed depending on the degradation of DNA.RNA hybrid duplex. This method can be introduced into the high throughput screening of the inhibitors against the RNase H activity for anti-HIV drug. PMID- 12903266 TI - Structural bases for substrate recognition and repair system of base-excision DNA repair proteins. AB - The model structure of Escherichia coli AlkA (3-methyladenine-DNA glycosylase II) protein complexed with the double helical DNA is elucidated from X-ray structures of related DNA glycosylase enzymes and mutagenic studies. The free enzyme structure has no difficulty in building the platform to afford the bended and wedge DNA with the flipped out nucleotide. The helix-hairpin-helix motif and the insertion residue L125 in free structure can be located without severe contacts. The alkylated base is surrounded with a variety of aromatic rings, such as W218, W272, Y273 and F18. The aromatic indole ring of tryptophan is a good candidate for forming the stacking with the positively charged base moiety pi-cation interaction). Some hydrophobic residues, such as V128 and L240, also attend to substrate recognition. PMID- 12903267 TI - Electrochemical characterization and DNA-binding property of dipyridophenazine complexes of osmium (II). AB - Novel dipyridophenazine (DPPZ) complexes of osmium (II), [Os(L)2(DPPZ)]2+ [L = 2,2'-bipyridyl (bpy)(1), 4,4'-diamino-2,2'-bipyridyl (DA-bpy)(2), 4,4'-dimethyl 2,2'-bipyridyl(DM-bpy)(3), and 4,4'-dicarboxyl-2,2'-bipyridyl (DC-bpy)(4)] have been synthesized and characterized. The DNA-binding properties of the complexes were studied by electrochemical methods. As the results, complex 2 shows higher affinity to DNA than other osmium complexes. The binding constant, K of complex 2 to calf thymus DNA has been determined to be 2.3 x 10(7) M-1 by normal pulse voltammetry (NPV). PMID- 12903268 TI - Thermodynamic analyses of triplex formation with homopurine oligonucleotide. AB - We analyzed the thermodynamics of purine motif triplex formation by isothermal titration calorimetry. The signs of calorimetric enthalpy change, delta Hcal, and entropy change, delta S, of the triplex formation were negative in the temperature range between 15 and 35 degrees C. Since an observed negative delta S was unfavorable for the triplex formation, the triplex formation was driven by a large negative delta Hcal. delta Hcal decreased with increasing temperature, yielding a negative heat capacity change, delta Cp, of approximately -1.2 kcal mol-1 K-1. We found that the binding constant, Ka, increased with increasing temperature, leading to an apparent positive van't Hoff enthalpy change, delta Hvh, which was in sharp contrast with the large negative delta Hcal. The analyses of the observed temperature dependence of Ka and delta Hcal and the negative delta Cp suggest that the purine motif triplex formation near room temperature is not a simple two-state binding process but exhibits multiple states, which was previously observed for the pyrimidine motif triplex formation near room temperature. PMID- 12903269 TI - Atomic force microscope of drug-DNA interaction. AB - We have been investigated the possibility of B-Z transition in ZnTMPyP-DNA interaction based on the observation of spectroscopic data. In this study, we found drastic change in the AFM image of supercoiled plasmid DNA when it was interacted with TMPyPs indicating that the considerable amount of unwinding of double helix or B-Z transition is induced by the drug-DNA interaction. Such phenomena were not observed for other cationic drugs examined. PMID- 12903270 TI - NMR study of a novel RNA quadruplex structure. AB - The structure of an RNA oligomer, r (GGAGGUUUUGGAGG) (R14-2) whose G-G steps are separated by adenine and uracil residues has been investigated by NMR. In the presence of 20 mM K+, a novel dimeric multiplex architecture is adopted by two strands of R14-2. In each strand a UUUU loop and two A residues connect four parallel G-G steps that pair-align into two tetrads. One of the tetrads is further pair-aligned by two A residues through the sheared mismatch and a novel hexad is subsequently formed. Two hexads coming from two different strands stack to make a dimeric multiplex. All of the guanosine and adenosine residues take an anti conformation. PMID- 12903271 TI - Separation of triplet repeat DNA by capillary electrophoresis and the conformational analysis by atomic force microscope. AB - We investigated the relationship between electrophoretic behaviors and higher order structures of triplet repeat DNA fragments by means of capillary electrophoresis and atomic force microscopy (AFM). It was suggested that the mobility difference between triplet repeat DNA and random sequence DNA should be correlated to differences in their dynamic conformation. PMID- 12903273 TI - Retrovirus-specific random mutagenesis by a nucleoside 5'-triphosphate analogue, PTP. AB - In a retrovirus replication model system, which consists of in vitro transcription and reverse transcription cycles, 6-(beta-D-ribofuranosyl)-3,4- dihydro-8H-pyrimido[4,5-c][1,2]oxazin-7-one-5'-triphosphate (PTP) induced highly efficient random mutations and this was due to the ambiguous incorporation of PTP by RNA polymerases. The types of mutations were mainly C-to-U or U-to-C transition mutations and the frequency was about 4 x 10(-2)/nucleotide during four cycles of the replication. Since a high mutation rate is harmful to species, PTP may be new candidate for anti-retroviral drugs. N4-aminoCTP and N4-hydroxyCTP were also incorporated ambiguously by RNA polymerase. These compounds may have a potential to induce mutation by the same mechanism as PTP. PMID- 12903272 TI - Random insertion and deletion mutagenesis for construction of protein library containing nonnatural amino acids. AB - A new method was developed for the generation of a library of mutant proteins that contained nonnatural amino acids. The method, "random insertion and deletion (RID) mutagenesis", is based on the deletion of an arbitrary number of bases at random positions and, at the same time, the insertion of an arbitrary sequence into the same position. By using this method, randomly selected three consecutive bases in the gene of green fluorescence protein (GFP) were replaced by a CGGT 4 base codon. When this DNA library was expressed in E. coli, about 80% of colonies lost the fluorescence. The non-fluorescent colonies were picked up and the genes were sequenced. Replacement of three consecutive bases by CGGT 4-base codon was found in two of the four mutated genes. PMID- 12903274 TI - DNA lesion and mutagenesis induced in phageM13mp2 by UVA, UVB and UVC irradiation. AB - Sunlight is carcinogenic and mutagenic and its genotoxic effects are believed to be the result of UV light-induced lesions in DNA. These lesions include pyrimidine dimers and (6-4) photoproducts, but it is uncertain whether the pyrimidine modifications are the sole pre-mutagenic lesions induced by UV light. Previous studies indicate that some sunlight-induced mutations in the single stranded DNA phage M13mp2 may not be caused by these photoproducts. In this work, purified single-stranded phage DNA was exposed to UVA, UVB and UVC and the induced mutations were analyzed. All 3 types of UV light increase the mutation frequency. The mutants were sequenced and the results suggest that UVA exposure may induce formation of a non-dipyrimidine lesion in DNA. PMID- 12903275 TI - Design and synthesis of curcumin-bioconjugates to improve systemic delivery. AB - Di-O-glycinoyl curcumin (I), di-O-glycinoyl-C4-glycyl-curcumin (II), 5'-deoxy-5' curcuminyl thymidine (5'-cur-T) (III) and 2'-deoxy-2'-curcuminyl uridine (2'-cur U) (IV) have been synthesized and characeterised by elemental analysis & 1H NMR. The antibacterial activities of these four bioconjugates has been tested particularly for multiresistant micro-organisms. Best results are shown by I & IV. These bioconjugates serve dual purpose of systemic delivery as well as therapeutic agents against viral diseases. PMID- 12903276 TI - Quantification analysis of translation initiation signal in vertebrate mRNAs: effect of nucleotides at positions +4(-)+6 upon efficiency of translation initiation. AB - Concerning the translation initiation signal in vertebrate mRNAs, a consensus, sequence, (GCC)GCC(A or G) CCATGG, has been proposed, but actual initiation sequences differ from it in a greater or lesser degree. Kozak monitored selection by ribosomes of the first versus second ATG codons as a function of mutations introduced at positions +4, and +6 of the first ATG codon. Codons possessing G at +4 strongly enhanced selection of the first ATG codon. However, ATG codon recognition was unaffected by most mutations in positions and +6. These data were well understood by our quantification analysis. PMID- 12903277 TI - Development of a novel functional biosensor with a short Ca(2+)-dependent deoxyribozyme. AB - We develop a novel functional biosensor on a deoxyribozyme. A 5'-end-immobilized short Ca(2+)-dependent deoxyribozyme (dCGCTGGCAGGCTACAACGAGTCTTC) binds to a target RNA substrate (rGAAGACA decrease UGCCAGCG; decrease denotes an RNA cleavage site), and acts as an enzyme in the presence of Ca2+. It cleaves the target RNA substrate at one site of rAp decrease U in the asymmetric internal loop. PMID- 12903279 TI - Threonyl-tRNA synthetase of archaea: importance of the discriminator base in the aminoacylation of threonine tRNA. AB - To investigate the contribution of the discriminator base of archaeal tRNA(Thr) in aminoacylation by threonyl-tRNA synthetase (ThrRS), cross-species aminoacylation between Escherichia coli and Haloferax volcanii, halophilic archaea, was studied. It was found that E. coli ThrRS threonylated the H. volcanii tRNA(Thr) but that E. coli threonine tRNA was not aminoacylated by H. volcanii ThrRS. Results of a threonylation experiment using in vitro mutants of E. coli threonine tRNA showed that only the mutant tRNA(Thr) having U73 was threonylated by H. volcanii ThrRS. These findings indicate that the discriminator base U73 of H. volcanii tRNA(Thr) is a strong determinant for the recognition by ThrRS. PMID- 12903278 TI - Mutagenicity of 5-formyluracil in mammalian cells. AB - 5-Formyluracil, a major oxidized form of thymine, was incorporated into a predetermined site of one of the leading and lagging template strands of a double stranded vector, and the DNA replication efficiency and the mutation frequency of 5-formyluracil in simian COS-7 cells were investigated. 5-Formyluracil did not block DNA replication and was weakly mutagenic in simian cells. 5-Formyluracil primarily elicited base substitutions at the modified positions. PMID- 12903280 TI - Novel fluorophore for labelling of oligonucleotides. AB - A novel fluorophore viz. 4-dansylamido-1,8-naphthalimido-N-pentanol has been designed, prepared and characterised. The comparative fluorescence has been studied in different solvents, solvent gradient, aqueous solutions of inorganic ions and buffers. This can be used for covalent tagging of oligonucleotides having potential application in Molecular Biology. PMID- 12903282 TI - Evaluation of nucleotide derivatives and sugar nucleotide analogs as inhibitors of glycosyltransferases. AB - The inhibition of glycosyltransferases was studied using uridine monophosphate derivatives and uridine diphosphate sugar analogs. Modification in the nucleoside portion caused selective inhibition of glycosyltransferases. PMID- 12903281 TI - Preparation and enzymatic recognition of guanine lesions induced by nitrogen oxides. AB - Xanthine (Xan) and oxanine (Oxa) are the major deamination products of guanine formed by the treatment with nitrogen oxides (e.g., NO and HNO2). In this study, 2'-deoxyribonucleoside 5'-triphosphates of Xan and Oxa were prepared by the NaNO2 treatment of dGTP. These modified nucleotides were incorporated into oligonucleotides by DNA polymerase reactions. The repair activities of various DNA N-glycosylases for Xan and Oxa were examined using these substrates. PMID- 12903283 TI - Discontinuous translation and mRNA structure of the coding region. AB - An analysis of the published data of the electrophoresis of the products of silk fibroin exhibiting discontinuities or pauses in the translational process showed that about 60 times of pauses occur during the translation of the mRNA from silk fibroin. In addition, we estimated that silk fibroin gene as a whole (15 kb) is composed of about 60 alternate, crystalline-noncrystalline, arrays of nucleotide sequence elements each connected by the boundary sequence, on the basis of the nucleotide sequence of the partial cDNA clone of the heavy chain gene of Bombyx mori available in the literature. The coincidence of the total number of pauses with that of the alternate arrays suggests that one translational pausing site exists in each of the unit (alternate array + boundary) nucleotide sequence in the silk fibroin mRNA template. PMID- 12903285 TI - Genome structure of Ri plasmid (3). Sequencing analysis of the vir region of pRi1724 in Japanese Agrobacterium rhizogenes. AB - The entire genome of the pRi1724 (217.6-kb) in the mikimopine type Agrobacterium rhizogenes strain MAFF03-01724 has been completely sequenced. The vir region covering 30.2-kb has found to be composed of 21 genes resembling virH1, virA, virB1-11, virG, virC1-2, and virD1-5. The structural organization of the pRi1724 vir operons in this study is exactly the same as that of the previously reported vir operons of other Ri or Ti plasmids, although the size of some ORFs showed little variations among the plasmids. We also found virE3 gene in the pRi1724 (1), but different from Ti plasmids, virE1 and virE2 that are also important for the virulence do not exist in the vir region of pRi1724. PMID- 12903284 TI - In vitro cleavage of Drosophila 2S rRNA by M1 RNA. AB - Drosophila melanogaster initiator methionine tRNA can adopt an alternative conformation in aqueous solution. In this alternative conformation, the aminoacyl and the anticodon stems of tRNA are unfolded and then these unfolded regions are used to form extended D- and T-stems, resulting in the formation of two tandemly joined stems and loops. This conformational alternation was recognized then cleaved by the catalytic RNA of Escherichia coli ribonuclease P (M1 RNA). The cleavage occurs within the mature sequence of tRNA. This further processing within mature sequence is called hyperprocessing. During the screening experiments of other conformational changeable D. melanogaster tRNAs by M1 RNA, we incidentally found that M1 RNA also hyperprocessed D. melanogaster 2S rRNA. Kinetic analyses of the hyperprocessing reaction of 2S rRNA by M1 RNA revealed that 2S rRNA could form a homodimer. PMID- 12903286 TI - Toward structural and functional genomics of Agrobacterium tumefaciens: linkage map of the left region of linear chromosome. AB - Genome of A. tumefaciens contains a linear and a circular chromosome. As an initial step of elucidating the structural and functional genomics of this bacterium, linkage map of the left region of its linear chromosome was constructed. Total genomic libraries of A. tumefaciens MAFF301001 were constructed in BAC vectors namely, pFOS1 and pBeloBAC11. Upon construction of sub libraries, minimum overlapping clones needed to cover the left region was determined. So far, four contigs have been assembled with a total of 19 overlapping clones. Detailed EcoRI physical map of contig III was constructed and it covers a 110 kb region of the Pme5 fragment of the linear chromosome. Seven end regions of the linking clones were partially sequenced but no gene existence was determined due to low homology. PMID- 12903287 TI - Incorporation of nonnatural amino acids into proteins by using five-base codon anticodon pairs. AB - A novel strategy for the incorporation of nonnatural amino acids into proteins was developed by using five-base codon-anticodon pairs. The streptavidin mRNA containing five-base codon CGGUA and the chemically aminoacylated tRNA with five base anticodon UACCG were prepared, and added into E. coli in vitro translation system. As a result, the nonnatural amino acid was successfully incorporated into desired position of the protein. Other five-base codons CGGN1N2, where N1 and N2 indicate one of four nucleotides, were also available for the incorporation of the nonnatural amino acid. PMID- 12903288 TI - Analysis of double-strand-break repair by Chlorella retrotransposon Zepp. AB - To elucidate the contribution of LINE-like retrotransposon Zepps in formation and maintenance of chromosomal telomeres, newly formed mini-chromosomes in irradiated Chlorella vulgaris cells were isolated and structurally characterized. A mini chromosome Y32 (approximately 400 kbp in size) was shown to have several copies of Zepp elements on both termini. On the right arm terminus, two copies of Zepps were found in a tandem array with poly(A) tracts facing towards the chromosome end. The poly(A) tail and a 3'-end of approximately 400 bp of the distal copy was replaced by telomeric repeats. On 5'-side of the proximal copy was another Zepp element in a reversed orientation. This newly formed telomeric structure is very similar to that found in the left arm terminus of chromosome I and support the model of Zepp-mediated maintenance of Chlorella telomeres. PMID- 12903289 TI - Introduction of a benzoyl group onto 6-chloropurine riboside in aqueous solution and its application to the synthesis of nucleoside derivatives. AB - A benzoyl group was introduced onto the 3'-hydroxyl group of 6-chloropurine riboside by treatment with benzoic anhydride in the presence of a base in aqueous solution. The product (3b) was converted to 9-(2,3-Di-deoxy-2-fluoro-beta-D-threo pentofuranosyl)adenine (1, FddA) in 6 steps, including radical deoxygenation. PMID- 12903290 TI - Synthesis of 4'-ethynyl-purine nucleosides possessing anti-HIV activity. AB - Searching for more effective anti-HIV agents, we have prepared 4'-ethynyl-purine nucleosides. They were derived in several steps from 4-C-triethylsilylethynyl ribose, which was used as an intermediate in the synthesis of pyrimidine nucleosides. The adenine derivative exhibited significant anti-HIV activity and favorable cytotoxicity profile in vitro. PMID- 12903291 TI - Lithiation study on D4T. AB - Upon treatment with LTMP, 5'-O-protected D4T undergoes deprotonation of the vinylic proton (H-3' or H-2'): when 5'-O-silyl derivative was used, the 3'-C silylated product was formed as a result of C3'-lithiation and subsequent O-->C silyl migration, while deprotonation at the 2'-position led to the formation of an allene derivative. A stannyl version of this reaction was also examined to develop a method for C3'-functionalization of D4T. PMID- 12903292 TI - Synthesis and radical reaction of 1'-phenylselenonucleosides. AB - The 1'alpha-phenylselenouridine derivative (4) was successfully synthesized via enolization at the 1'-position of the 3',5'-O-TIPDS-2'-ketouridine (1). After the introduction of a vinylsilyl tether as an intramolecular radical acceptor at the 2'-hydroxy group of 4, its atom-transfer radical cyclization reaction, followed by the treatment with TBAF gave 1'alpha-vinyluridine derivative (10). Using this procedure, 1'alpha-vinyluridine (11) and -cytidine (14) were successfully synthesized. PMID- 12903294 TI - Synthesis of 2,3-dihydroinosine derivatives by reduction using BH3-THF. AB - A novel reductive method for the chemical modification of nucleosides is described. Reaction of inosine derivatives with boran-THF resulted in the regioselective reduction of purine ring to afford the corresponding 2,3 dihydroinosine derivatives in moderate yields. PMID- 12903293 TI - Synthesis of carbocyclic nucleosides and their SAH hydrolase inhibitory activities. AB - The cellular enzyme S-adenosyl-L-homocysteine (SAH) hydrolase has emerged as a target enzyme for the molecular design of anti-viral agents. Recently, SAH hydrolase has been considered as an attractive target in parasite chemotherapy for malaria. We report synthesis of several carbocyclic purine nucleosides and their inhibitory activities against human and malaria recombinant SAH hydrolases. PMID- 12903296 TI - Synthesis and chirality analysis of carbocyclic 5'-nor nucleosides. AB - Achiral carbocyclic "DL-like" 5'-nor nucleosides have been synthesized and analyzed by the chiral capillary electrophoresis to elucidate the "D-like" monomers. PMID- 12903295 TI - Enantiodifferentiating Z-E photoisomerization of cyclooctene sensitized by DNA and RNA. AB - DNA and RNA have been shown for the first time to function as chiral photosensitizers in aqueous solution, to effect the enantiodifferentiating photoisomerization of (Z)-cyclooctene (1Z), giving the chiral (E)-isomer in enantiomeric excesses (ee's) of up to 15%. In order to elucidate the effect of nucleotide sequence, enantiodifferentiating photoisomerization of 1Z was also performed using oligo and homopolynucleotides as chiral sensitizer. The -18.8% ee was observed by using d(T)15.d(A)15 as sensitizer, whereas sensitization by the poly(U).poly(A) duplex gave only racemic (E)-cyclooctene. From these results, oligothymidine sequence is essential for efficient enantioselective photoisomerization of 1Z. PMID- 12903297 TI - New method for scanning of single-nucleotide polymorphisms (SNPs): recognition of guanine-guanine mismatches by dimeric naphthyridine. AB - Dimeric naphthyridine was designed and synthesized as a novel ligand that specifically binds to G-G mismatch, one of four SNP types. In the presence of dimeric naphthyridine, CD spectra of the G-G mismatch containing duplex noticeably changed, being accompanied by the induced CD at 300-350 nm, whereas no CD spectral change was observed for normal duplex. DNaseI footprinting titration indicated a selective binding of dimeric naphthyridine to the G-G mismatch with a dissociation constant of 53 nM. PMID- 12903298 TI - Synthesis of a thymine glycol building block and its incorporation into oligonucleotides. AB - A phosphoramidite building block of thymine glycol, a major oxidative base damage in DNA, was synthesized in four steps from protected thymidine. The two hydroxyl functions of the oxidized base were protected with the tert-butyldimethylsilyl group. Using this building block, oligonucleotides containing this damage were synthesized. PMID- 12903299 TI - Solid-phase synthesis of deoxyribonucleotides having a cap structure analog. AB - In this study, the solid-phase synthesis of oligodeoxyribonucleotides having a guanosine pyrophosphate cap structure (Gpp-) was achieved by using a new guanosine monophosphate unit having the DMTr group capable of estimation of coupling efficiency of the pyrophosphate bond formation. Since 7-methylguanosine base was unstable under basic conditions, Gpp-capped DNA oligomers were synthesized by using a new linker having a silanediyl bond, which allowed to release the DNA chain from the solid support by treatment with fluoride anion under neutral conditions. PMID- 12903300 TI - Synthesis and properties of a new type DNA dendrimer. AB - We have designed a new type of a DNA dendrimer which has rigid branched structure. The branching molecule was prepared from 1,3,5-tribromobenzene. The dendrimer unit, in which three oligonucleotide-chains, two molecules of T15 and one molecule of A15, linked to the branching molecule, was synthesized by an automated DNA synthesizer. The properties of the dendrimer unit and dendrimer formation by inter-molecular association of T15 and A15 chains of the dendrimer unit will be presented. PMID- 12903301 TI - Synthesis of oligodeoxyribonucleotides containing 2'-deoxypseudouridine: inhibition of uracil-DNA glycosylase. AB - 2'-Deoxypseudouridine (1) was synthesized starting from thymidine in a 23% overall yield via a silylated furanoid glycal. Compound 1 was introduced to oligodeoxyribonucleotides (ODNs) by the solid phase phosphoramidite chemistry. The ODNs containing 1 inhibited uracil-DNA glycosylase (UDG) reaction. PMID- 12903302 TI - Efficient cross-linking to cytidine using substituted phenylsulfide derivatives of 2-amino-6-vinylpurine nucleoside via synchronous activation within duplex. AB - We have previously described that oligonucleotides containing phenylsulfoxide derivative of 2-amino-6-vinyulpurine nucleoside analog (1) are activated within duplex to form cross-link toward cytidine selectively at the target site. The new cross-linking motif with phenylsulfoxide structure (2) is characteristic in that the stable precursor may be transformed automatically within duplex to a reactive species. To search for more stable precursor susceptible for activation, we designed a series of substituted phenylsulfide analogs of 1. It has been demonstrated that introduction of an electron-donating group on the phenyl ring improved the cross-linking reaction. Particularly, 2-carboxyphenyl sulfide derivative exhibited cross-linking as effectively as phenylsulfoxide derivative without chemical oxidation prior to cross-linking. PMID- 12903303 TI - Synthesis and triplex forming ability of conformationally locked oligonucleotides containing unnatural nucleobases: efficient recognition of a C.G interruption. AB - In order to develop a novel nucleoside analogue which recognizes C.G interruption in homopurine.homopyrimidine DNA, we designed and synthesized a conformationally locked nucleoside analogue, 1-(2-O,4-C-methylene-beta-D-ribofuranosyl)pyridin-2 one (4), and introduced it into a triplex-forming oligonucleotide (TFO). On melting temperature (Tm) measurements, the unprecedented C.G base recognition ability of 4 was observed. PMID- 12903304 TI - Novel synthesis of a tetra-acridinyl peptide as a new DNA polyintercalator. AB - Tetra-acridinyl peptide 2 was synthesized from Fmoc-Lys(Boc)-OH and Fmoc-Lys(Acr) OH (1) with the peptide synthesizer. The CD measurement suggested that 2 forms a special organized structure by itself in buffered solution. Peptide 2 binds to double stranded DNA with a very large affinity constant, which is 10(3)-times larger than that of quinacrine. Spectrophotometric and hydrodynamic studies suggested that all of the acridinyl parts of 2 contribute to the intercalating interaction for the DNA binding. Our finding in this experiment demonstrates that polyintercalators such as 2 can be assembled quickly by the automated synthesizer. PMID- 12903305 TI - A novel approach to the synthesis of oligodeoxyribonucleotide boranophosphates. AB - Deoxyribonucleoside 3'-boranophosphate derivatives including adenine, cytosine, guanine, and thymine bases were synthesized in good yields by the use of a new boranophosphorylation reaction. The reaction was found to be effective for the formation of internucleotidic boranophosphate linkages. PMID- 12903306 TI - An efficient synthesis of nucleotides via the phosphoramidite method using a triflic acid salt of an imidazole-related compound as a promoter. AB - N-Phenylimidazolium triflate and N-methylbenzimidazolium triflate, new imidazole related compound/triflic acid-complex type of promoters in the phosphoramidite method, has been developed. These reagents are, particularly, useful for internucleotide-bond formation with lowly reactive reactants and have allowed an efficient, high-yield synthesis of oligodeoxyribonucleotides both in a solution phase and on a solid supports. PMID- 12903307 TI - GC-->CG transversion mutation might be caused by 8-oxoguanine oxidation product. AB - Oxidation of d(A6GGA6)/d(T6CCT6) with sensitizer under different UV irradiation periods was examined. Enzymatic digestion of photolysate revealed that the amount of 8-oxoG initially increased with decrease of G and then decreased with the formation of Iz. In order to investigate whether Iz was formed by one electron oxidation of 8-oxoG through long-range hole migration, photoirradiation of anthraquinone-linked oligomer was examined. The results clearly demonstrated that 8-oxoG was efficiently converted to Iz under irradiation conditions. These results suggest that Iz generated from 8-oxoG induces GC->CG transversion under oxidative condition. PMID- 12903308 TI - Design of an artificial restriction enzyme having simultaneous DNA cleavage activity. AB - We designed dimeric distamycin linked hydroxamic acid (DDHA)-metal complexes and investigated double strand simultaneous cleavage. Compared as distamycin linked hydroxamic acid (DHA)-metal complex, DDHA-metal complex cleaved DNA at the same level of DHA-metal complex without double strand simultaneous cleavage. PMID- 12903309 TI - Synthesis of non-natural DNA using DNA polymerase. AB - Nonnatural nucleotide modified by glucose or galactose was synthesized to increase functional diversity of DNA library. These compounds were incorporated in a DNA double strand using Klenow Fragment as well as dTTP. These functional group could be ordered sequentially on a DNA double strand at intervals of few angstroms according to the designed template sequence within a few hours. This method must be useful to constructing nonnatural DNA library or designed supramolecular structures. PMID- 12903310 TI - Measurement of the time course of DNA/RNA hybridization and RNA detection using fluorescence polarization analysis. AB - Using fluorescence polarization analysis, the time courses of hybridization between probe oligo-DNAs and target RNAs were measured. The RNAs were amplified using the DNA templates of Shiga toxin genes by NASBA (Nucleic Acid Sequence Based Amplification). Two DNA probes were designed for detecting the genes and they rapidly and specifically hybridized with their target RNA sequences. NASBA could be sufficiently used for the combination and DNA/RNA hybridization could be detected in the fluorescence polarization. PMID- 12903311 TI - Mechanistic studies of RNA modifying enzymes. RNA pseudouridine synthase and m5cytosine methyl transferase. AB - RNA has numerous post-transcriptional modifications, but relatively little is known about the enzymes that catalyze such modifications or about the functions of the modified residues. Our laboratory has been engaged in studies of the structure and function of enzymes that catalyze the conversion of Urd residues in RNA to pseudouridine (psi Synthase), and Cyd to 5-methylcytidine methyl transferase (RNA m5C Mtase). The presentation will summarize recent results from our laboratory. PMID- 12903312 TI - A study of the method to pick up a selenocysteine tRNA in Bacillus subtilis. AB - In Bacillus subtilis, selenocysteine tRNA has not been identified in a total genome sequence so far (1). To explore the system of selenocysteine incorporation in B. subtilis, we screened serine-acceptable tRNAs to find an unknown tRNA for selenocysteine by the combined method of specific biotinylation of aa-tRNA (2) and RT-PCR (3). cDNAs obtained from the serine-acceptable tRNA pool were amplified and cloned into plasmid to read its sequence. This procedure gave cDNA library corresponding known serine tRNAs, but no candidate for selenocysteine has been found. Thus, this result, together with the previous data (4), might reveal that there is no selenocysteine tRNA in B. subtilis and/or metabolism of selenium is considerably different from known one as seen in other bacteria. PMID- 12903313 TI - Complex formation of double-stranded DNA (6-4) photoproducts and anti-(6-4) photoproduct antibody Fabs. AB - DNA (6-4) photoproducts are major constituents of ultraviolet-damaged DNAs. We prepared double-stranded (ds) (6-4) DNA photoproducts and analyzed formation of their complexes with anti-(6-4) photoproduct antibody Fabs. Elution profiles of the mixtures of ds-(6-4) DNAs and Fabs from anion-exchange and gel-filtration columns indicate that Fab 64M-2 deprives 14mer ds-(6-4) DNA of single-stranded (ss) (6-4) DNA and shows no interaction with 18 mer ds-(6-4) DNA (A18). Fab 64M-5 with an approximately 100-fold higher affinity than Fab 64M-2 forms a complex with the ds-(6-4) DNA (A18), but partly dissociates another 18 mer ds-(6-4) DNA (A18-3), with a lowered G-C content, into ss-DNAs. From these results, antibody 64M-5 possibly accommodates the T(6-4)T photolesion moiety of the ds-(6-4) DNA (A18) by flipping out the moiety from its neighboring segments. PMID- 12903314 TI - Fluorometric assay of DNA binding proteins using class IIS endonuclease. AB - Fluorometric DNA-protein binding assay was developed with class IIS restriction enzyme and FITC-labeled DNA fixed on magnetic beads. We used this assay with AP1 and FokI as a case study to quantify DNA-protein binding. Fluorescent images of the treated beads showed that the transcriptional factor bound to its DNA site and inhibited the restricted digestion effectively. The affinity of AP1 to its site was estimated as a Kd value, which was approximately 10 nM. PMID- 12903315 TI - Cloning and sequencing of ftsZ homolog from extremely halophilic archaeon Haloarcula japonica strain TR-1. AB - The gene encoding FtZ was cloned from triangular disc-shaped extremely halophilic archaeon Haloarcula japonica strain TR-1. Nucleotide sequencing analysis of the possible ftsZ gene revealed that the structural gene consisted of an open reading frame of 1,182 nucleotides encoding 394 amino acids. The deduced amino acid sequence of the Ha. japonica FtsZ showed high identities with those Halobacterium salinarom, Haloferax volcanii and Haloferax mediterranei FtsZs. PMID- 12903316 TI - A novel method for DNA molecular counting. AB - We have developed a new method for counting DNA molecules using 'capillary plates' consisting of a large number of small glass-capillary 'channels' fused together in parallel. PCR mixtures containing serially diluted DNA templates with the DNA indicator dye Hoechst 33258 were poured into the plates and sealed with silicone rubber-plates. Following 40 PCR cycles, fluorescence microscopy revealed that the fluorescence in some channels had increased about three-times more than in others at template concentrations of 1 fM or lower. No bright fluorescence was observed in the absence of template. The relationship between the proportion of fluorescent channels in the capillary-plates and the template concentrations was linear according to Poisson probabilities in the range of 0.1-1,000 aM. These results demonstrate the amplification of single templates in the channels, and that a small number of templates could be quantified by counting the proportion of positive channels on the capillary-plates. PMID- 12903317 TI - Improvement of IGCR technique using FRET. AB - The fluorescent detection system has been introduced into the study on denaturation/reassociation process of DNA fragments in gel, for improving In-Gel Competitive Reassociation technique, one of genome subtraction methods. The annealing behaviour of the mixture of 3'-Fluorescein-labelled and 5'-Cy5-labelled DNA fragments was analysed by Fluorescence Resonance Energy Transfer (FRET) technique from donor Fluorescein to acceptor Cy5. We showed that two fluorescent dyes labelled at 3' and 5' ends of DNA fragments caused FRET in both the solution and the gel. The characterisation of fluorescence-labelled fragments in gel and the changes of their fluorescence intensity will be reported. PMID- 12903318 TI - Characterization of immediate early genes expressed in chlorovirus infection. AB - By Southern blot analysis of restriction fragments of a chlorovirus CVK2 genomic contig with probes of RNA expressed immediate early in infection, sixteen genes were specifically found to be expressed in the host cells. These genes include those for aminoacyl-tRNA synthetase, nucleolin, ribosomal protein S5, hyaluronan synthase, TFIID etc. All of these transcripts were polyadenylated and most likely expressed in the host nucleus. The structural characteristics of these genes are discussed in connection with their expression mechanism. The biological importance of the gene products in viral infection are also considered. PMID- 12903319 TI - Analysis of interaction between RNA aptamer and protein using nucleotide analogs. AB - Non-structural protein 3 (NS3) derived from Hepatitis C virus (HCV) is essential for viral proliferation and has two functional domains; trypsin-like serine protease and helicase. Recently we obtained three types of RNA aptamers (G9-I, II and -III) bound to NS3 protease domain (delta NS3) by in vitro selection and confirmed their strong inhibition for protease activity. These aptamers have a common sequence, 5'-GA(A/U)UGGGAC-3', forming a loop structure by Mulfold secondary structure modeling. G9-I shows a three-way junction and G9-II and -III have four-way junction structures. To characterize the active structure of these aptamers, we applied modification interference analysis using nucleotide analogs and identified common important nucleotides in these three aptamers. PMID- 12903320 TI - Phage display of xylan-binding module of xylanase J from alkaliphilic Bacillus sp. strain 41M-1. AB - Xylanase J from alkaliphilic Bacillus sp. strain 41M-1 has a family 11/G catalytic domain and a xylan-binding module (XBM). The XBM of xylanase J was displayed on the surface of filamentous bacteriophage. The XBM expressed on the phage surface retained binding activity to xylan. Random mutations were introduced in the XBM gene by error-prone PCR, and the repertoire was cloned for display on phage. Sequence analysis of the xylan-binding activity-deficient mutants revealed that Phe 284 and Trp317 of the XBM would contribute to the xylan binding activity. PMID- 12903321 TI - Identification of Thermus thermophilus HB8 tRNA (Gm18) methyltransferase gene. AB - For the purpose of identification of the gene for Thermus thermophilus tRNA (Gm18) methyltransferase [tRNA (guanosine-2'-)-methyltransferase, EC 2.1.1.34], the purified enzyme from native source was analyzed by the peptide-mass mapping. The target gene encoded the amino acid sequences of the obtained peptides was searched in data from Thermus thermophilus HB8 genome-sequencing project. We found the target gene AB05130, which was expected to encode a protein composed of 194 amino acid residues and the molecular mass of this protein was calculated as 22083. The recombinant protein was expressed in E. coli as an active form. The Gm18 formation activity of the purified recombinant protein was confirmed by in vitro methylation followed by two-dimensional thin layer chromatography and Liquid Chromatography/Mass Spectrum analysis of substrate tRNA. PMID- 12903322 TI - Functional analysis of the pro-apoptotic factor Bax using hammerhead ribozymes. AB - A pro-apoptotic protein Bax is a Bcl-2 family member and forms homodimers and also heterodimerizes with death antagonists, Bcl-2 and Bcl-XL. To elucidate the detail of function of Bax in cells, we constructed a hammerhead ribozyme targeted to the Bax mRNA. The level of Bax protein in Hela-K cells expressing Bax-ribozyme was decreased compared with that of wild type Hela-K cells. Therefore, the Bax ribozyme should be useful for the future investigations of the details of apoptosis pathway. PMID- 12903323 TI - Ferroceneacetyl naphthalene diimide as a new electrochemical ligand for DNA sensing. AB - New electrochemically active DNA ligand 1 was synthesized by the connection of ferroceneacetic acid at the terminal amino moieties of two imide substituents of naphthalene diimide. Electrochemical experiments in aqueous solution containing DMSO showed that the selectivity for double stranded DNA of 1 has increased from that of ligand 2 previously reported. The peak current of 1 shifted toward the negative side from that of 2, thereby shortening the time required for gene detection. PMID- 12903324 TI - Fluorescence microscopic visualization of a DNA-cationic fullerene complex. AB - Cationic fullerene derivative 1, which is soluble in aqueous medium, bound to double stranded DNA. Observation of Coliphage T4 DNA in the absence and presence of 1 by fluorescence microscopy suggested that 1 bound to DNA along its groove. Upon irradiation of visible light on the DNA complex of 1, DNA was cleaved dramatically. The process of DNA photo-cleavage could be monitored continuously by fluorescence microscopy. PMID- 12903325 TI - Inhibition of human telomerase by nucleotide analogues bearing a hydrophobic group. AB - Telomerase, which synthesizes telomeric DNA in eukaryotic cells, is classified as a reverse transcriptase. To clarify the susceptibility of telomerase to nucleoside 5'-triphosphates bearing a hydrophobic group on the base moiety, we studied the inhibitory effects of 2',3'-dideoxy-5-styryluridine 5'-triphosphate analogues and 9-(beta-D-arabinofuranosyl)-2-(p-n-butylanilino)purine 5' triphosphate analogues on telomerase activity using a quantitative 'stretch PCR' assay. 2',3'-Dideoxy-5-styryluridine 5'-triphosphate (StddUTP) showed more potent inhibition than 2',3'-dideoxythymidine 5'-triphosphate (ddTTP). On the other hand, 9-(beta-D-arabinofuranosyl)-2-(p-n-butylphenyl)guanine 5'-triphosphate (BuParaGTP) showed no inhibition, even though 9-(beta-D-arabinofuranosyl)guanine 5'-triphosphate (araGTP) is a potent inhibitor of telomerase. The influence on telomerase of hydrophobic substituents on the base moieties of nucleotides is described. PMID- 12903326 TI - Binding activity of replication protein A to UV-damaged single-stranded DNA. AB - To obtain the information for the exact role of replication protein A (RPA) on both eukaryotic DNA replication and repair, the binding preference of RPA purified from Xenopus egg extract against the undamaged and UV-damaged single stranded DNA was studied by the gel shift assay. Chemically synthesized oligonucleotide containing the pyrimidine(6-4)pyrimidone photoproduct at one site was used as a model of UV-damaged DNA. Results of competition assay and Scatchard plots indicate that RPA preferentially binds to the 6-4 photoproduct oligonucleotide than the undamaged DNA. PMID- 12903327 TI - Synthesis and evaluation of novel bioconjugates as antiviral agents. AB - Novel lipid (mannito-stearate) antiviral nucleoside and oligonucleotide conjugates were prepared with improved lipophilic and membrane associating properties of drugs. Potential advantages of these liponucleotide prodrugs are lower toxicity, increased cellular uptake, nuclease resistivity and antiviral activity. Oligonucleotide conjugate complementary to a unique segment of viral genome may selectively disrupt the processes dependent on the segment by hybridisation. PMID- 12903329 TI - Trans-kingdom conjugation between bacterial chromosome-less mini-cells and yeasts. AB - We introduced a novel convenient method to prepare chromosome-less mini-cells from cell mixture of Escherichia coli P678-54 by using penicillin. The mini-cells prepared by this method could successfully transferred their harboring conjugative plasmids into eukaryotic yeast cells by trans-kingdom conjugation. PMID- 12903328 TI - Properties of quadruplex oligonucleotides with anti-HIV-1 activity. AB - A phosphorothioate oligonucleotide composed of deoxyguanosine and thymidine was identified as an inhibitor of HIV-1 infection at an early stage of the HIV-1 replication cycle in vitro. The phosphodiester and phosphorothioate chimeric oligonucleotides were found to be potent inhibitors of several steps of HIV-1 infection: the interaction with CD4 and the chemokine receptor, the reverse transcriptase activity, and the integrase activity. To elucidate the mechanism of the anti-HIV-1 function of these oligonucleotides in terms of their structure, we focussed on their G-serial sequences and investigated the characteristics of their solution structures. These oligonucleotides were proved to be able to adopt G-quadruplex structures by UV and CD measurements. We presume that the anti-HIV-1 activities of these oligonucleotides are consequently attributable to G quadruplex formation. PMID- 12903330 TI - Different toxicity of Escherichia coli and Agrobacterium tumefaciens against tobacco BY2 cells. AB - Tobacco BY2 cells were withered away by the physical contact with intestinal bacterium Escherichia coli. In contrast, plant pathogenic bacterium Agrobacterium tumefaciens with/without tumor inducing Ti plasmid was not so toxic as E. coli. Mini-cells of E. coli decreased their toxicity. PMID- 12903331 TI - In vitro selection and analysis of RNA aptamer recognize arginine-rich motif (ARM) model peptide on a QCM. AB - To study RNA-peptide interactions, we performed an in vitro selection of RNA on a 27 MHz quartz-crystal microbalance (QCM) on which a simple R5 helix peptide was immobilized as a model of N peptide from bacteriophade lambda. The consensus sequences including a GNRA tetraloop were obtained from a random RNA pool after the 7th cycle selection. PMID- 12903332 TI - A graphic tool for circular genome maps. AB - A program has been developed for drawing map of circular DNA such as organelle or plasmid genome. Total size of the genome, gene names and positions, and other details, if required, should be prepared in a simple format text file then the program process it to a PostScript(R) (PS) file with which you can print a image of the map on suitable device(s). The final touch on the map can be given through editing the PS file. PMID- 12903333 TI - Homogenous simultaneous detection of HCV RNA and internal control by two-color fluorescence real-time monitoring of isothermal sequence amplification with INAF probes. AB - We demonstrated the homogenous simultaneous detection of HCV RNA and internal control by two-color fluorescence real-time monitoring of isothermal sequence amplification with INAF probes. PMID- 12903334 TI - Cytotoxic mechanisms of inhibitor of RNA synthesis, 1-(3-C-ethynyl-beta-D-ribo pentofuranosyl)cytosine (ECyd, TAS-106). AB - We investigated the molecular mechanisms of cell death induced by 1-(3-C-ethynyl beta-D-ribo-pentofuranosyl)cytosine (ECyd, TAS-106: Figure 1), a potent inhibitor of RNA synthesis, using mouse mammary tumor FM3A cells and human fibrosarcoma HT1080 cells. ECyd induced the characteristics of apoptosis on these cells, such as morphological changes, DNA fragmentations and caspase-3-like protease activation. General caspases inhibitor (Z-Asp-CH2-DCB) inhibited cell death. Interestingly, we also found that ECyd induced rRNA fragmentation with the size of 3.2, 2.8 and 1.5 kb, and which might be caused by inhibition of RNA synthesis. rRNA fragmentation was mainly occurred in D8 domain of 28 S rRNA, and the end of 5'-terminal sequence of 1.5 kb fragment was C3220pC3221p or C3221pG3222p, that was identical to the recognition sequence of RNase L. Furthermore, the fragmentation patterns of rRNA digested with RNase L resembled that of ECyd treated cells in shape. These results indicate that antitumor mechanisms of ECyd are involved in activation of RNase L. rRNA fragmentation may be one of the death events as a result of inhibition of RNA synthesis and play an important role in the antitumor activity of ECyd. PMID- 12903335 TI - The behavior and effect of isopoly (S-carboxymethyl-L-cysteine) derivatives of nucleic acid bases. AB - Isopoly(S-carboxymethyl-L-cysteine) derivatives of nucleic acid bases were prepared as antisense compounds. These compounds in vitro have been found to form stable complex with oligo-DNA or RNA. This paper deals with effect of antisense compounds in vivo. The target in this paper is the sequence of the PSD-95 protein linked with NMDA receptor. Excess passing of calcium ions through the loss of the signal pathway without PSD-95 proteins caused by antisense compound. The cells detailing with L-cysteine derivatives showed the lowest percentage of 19.1%. The data were compared with that of phosphotioate antisense compound. PMID- 12903336 TI - Analysis of the P7 region within the catalytic core of the Tetrahymena ribozyme by employing in vitro selection. AB - The highly conserved P7 region is generally believed to act as a major portion of the catalytic site in the Group I intron ribozyme. However, its functions have not been elucidated except for the fact that it specifically binds a cofactor guanosine required for self-splicing reaction. We attempted an in vitro selection experiment to determine the sequence requirements of this region in the mechanism of catalysis by using the Tetrahymena ribozyme. We found that the selected active clones have the secondary structure similar to that of the wild type with few exceptions. However, their primary sequences were not conserved except G264 and C311 that are the major elements of the binding site for the guanosine. Our results suggest that the unique secondary structure of the P7 region is a primary requisite for the catalytic function of this class of ribozymes. PMID- 12903337 TI - Characterization of RNA structure by bis-pyrene-labeled 2'-O methyloligonucleotides. AB - Properties of 2'-O-methyloligoribonucleotides containing two consecutive 2'-O-(1 pyrenylmethyl)uridine were investigated as a fluorescent probe to search the single strand regions of RNA. The bis-pyrene-labeled 2'-O methyloligoribonucleotide (OMUpy2) induced the formation of pyrene dimer upon hybridization with the complementary oligoribonucleotides and showed remarkable appearance of broad structureless fluorescence at 480 nm. Contrarily, when OMUpy2 was hybridized with the complementary oligodeoxyribonucleotides, such enhancement of fluorescence was scarcely observed. When various OMUpy2 were applied to E. coli 5S-rRNA, the fluorescence intensity at 480 nm was varied in a sequence specific manner. PMID- 12903338 TI - Crystallization of the most active RNA-cleaving deoxyribozyme. AB - The structural compositions of the most active deoxyribozyme and its derivatives have been examined by electrophoresis, and their crystallization conditions were surveyed for X-ray analysis. It has been found that Mg2+ ion is essential to form the active binary complex between the catalytic DNA and the substrate, and that heat-treatment is effective to prevent formation of the inactive quaternary complex between the two enzymes and the two substrates. Crystals obtained by the hanging drop vapor diffusion method are composed of the active binary complex. PMID- 12903339 TI - Construction of a ribozyme-expression system that effectively transports ribozymes to the cytoplasm. AB - In our previous studies, it was demonstrated that the activity of a ribozyme in vivo was governed by several parameters, which include a high level-expression of ribozyme, the intracellular stability of the ribozyme and colocalization of the ribozyme with its target RNA in the same cellular compartment. To generate ribozymes with significant activity in vivo, we have developed a ribozyme expression system based on a human tRNA(Val) promoter. Our tRNA-embedded ribozymes produced by our ribozyme-expression system remain relatively stable in cultured cells with half-lives longer than 30 min. Moreover, tRNA-ribozymes with a cloverleaf structure were efficiently exported from the nucleus to the cytoplasm, where they would effectively cleave target RNAs. In the present study, we investigated the relationship between the secondary structure of the tRNA ribozymes and the transport efficacy of them in mammalian cells by using a screening system in vivo. Furthermore, we also investigated the mechanism of the export of tRNA-embedded ribozymes both in mammalian cells and in Xenopus oocytes. PMID- 12903340 TI - Expression mechanism of the allosteric interactions in a ribozyme catalysis. AB - The contribution of substrate binding to cooperative regulation in the rate process of ribozyme catalysis has been investigated using allosteric ribozymes. The high sensitivity to the substrate lengths is attributed to the catalytic core folding which proceeds due to the energetic contribution of the substrate binding. One role of the effector (FMN) is the promotion of the core folding through the stabilization of the aptamer domain. Another role is the inhibition of the cleavage chemistry by perturbing the intermediate state in the rate process. The total effects of these two types of kinetic regulation determine the substrate dependency of the cooperative interaction on the catalytic reaction. An adequate correlation between the type of regulation and the substrate binding is responsible for the cooperative interaction in the kinetic process. PMID- 12903341 TI - Effective suppression of HIV-1 gene expression by a mammalian tRNA 3' processing endoribonuclease and external guide sequence oligozymes. AB - We examined the suppression of virus expression by cleavage of the HIV-1 RNA gene using a mammalian tRNA 3' processing endoribonuclease and an External Guide Sequence Oligozyme (EGS) in vivo. We constructed an EGS expression vector that used the tRNA(met) promoter as an expression cassette for EGS. The EGS expression vector was targeted to the upstream region of gag, region. The EGS expression vector was co-transfected into COS cells with the HIV-1 gene plasmid vector. As compared with the EGS non-expressing cells and the EGS expressing cells, the EGS expressing cells with the targeted gag start codon had a clearly decreased amount of the HIV-1 gag p24 protein. The EGS expressing cells with the targeted gag start codon showed effective suppression of HIV-1 gene expression. Thus, these studies describe novel gene targeting agents for the inhibition of gene expression and antiviral activity. PMID- 12903342 TI - Triplex formation using ODN conjugates with polycation comb-type copolymer. AB - Polycation comb-type copolymer that is composed of polylysine backbone and dextran side chains (PLL-g-Dex) has previously been shown to stabilize duplex and triplex DNAs quite effectively. In this study, we have conjugated PLL-g-Dex with oligonucleotides (ODN) aiming to increase the triplex stabilizing efficiency of the copolymer. Here we have demonstrated that the copolymer-TFO conjugates selectively stabilize triplex DNA. Also its potential to form triplex DNA was found to be greater than PLL-g-Dex/ODN mixture. PMID- 12903343 TI - Synthesis and conformation control of peptide ribonucleic acid containing 5' amino-5'-deoxyribopurinenucleosides. AB - A novel nucleic acid model, i.e. peptide ribonucleic acid (PRNA), tethering 5' amino-5'-deoxypyrimidine ribonucleoside as a recognition site for nucleic acids, has been designed and synthesized. We have demonstrated that the recognition behavior of PRNA with complementary oligopurinenucleotides can be controlled externally through the orientational switching of the pyrimidine nucleobase of PRNA induced by added borates. We extend this methodology of controlling the nucleobase orientation and recognition behavior of novel mono and oligomeric PRNAs containing 5'-amino-5'-deoxypyrimidine and/or purinenucleosides. In case of the PRNA oligomer containing pyrimidine-purine mixed sequence, efficient orientational switching of nucleobases induced by added borates was also observed. PMID- 12903344 TI - RecA-double stranded DNA complexes studied by atomic force microscopy. AB - RecA-double stranded (ds) DNA complexes have been studied by atomic force microscopy (AFM). When the complexes were prepared in the presence of ATP gamma S, fully covered RecA-dsDNA filaments were observed by AFM. When the concentration of RecA proteins was lower, various lengths of filaments were found. The variation of the observed structures may directly reflect the real distribution of the intermediate complexes in the reaction mixture, as the mixture was simply deposited on a mica surface for AFM observation without special fixation or staining. The use of a carbon nanotube (CNT) AFM tip enabled high resolution to reveal the periodicity of RecA-dsDNA filaments. Our observations demonstrated the potential of the AFM method for the structural studies of the RecA-dsDNA complexes, especially their intermediate states. PMID- 12903345 TI - Method for stretching DNA molecules on mica surface in one direction for AFM imaging. AB - An efficient method was developed to stretch DNA molecules on an atomically flat surface for AFM imaging. This method involves anchoring DNA molecules from their 5' ends to amino silanized mica surfaces. N-Succinimidyl6-[3'-(2-pyridyldithio) propionamido]hexanoate (LC-SPDP), a heterobifunctional cross-linker with a flexible spacer arm was used for this purpose. The immobilization process was carried out by introducing a thiol group to the 5' end of DNA by PCR. Thiolated molecules were then reacted with the cross linker to conjugate with its 2-pyridyl disulphide group via sulfhydryl exchange. The resulting complex was deposited on amino silanized mica where NHS-ester moiety of the cross linker reacted with the primary amino group on the surface. Samples were washed by a current of water and dried by an air jet in one direction parallel to the surface. DNA molecules were shown to be fully stretched in one direction on imaging them by AFM. PMID- 12903346 TI - Condensation reaction of hexanucleotides containing guanine and cytosine with water soluble carbodiimide. AB - The condensation reactions of hexanucleotides involving guanine and cytosine in the presence of water-soluble carbodiimide (WSC) have been investigated as a model reaction of the prebiotic formation of RNA under primitive earth. The reactions formed cyclic hexanucleotides and dimers in which the product yields were dependent on the sequence. PMID- 12903347 TI - Automated in vitro selection to obtain functional oligonucleotides. AB - In vitro selection or systematic evolution of ligand by exponential enrichment (SELEX) has been devised for the identification of high-affinity oligonucleotide aptamers to target molecules. However, the selection process is repetitive and time-consuming. We have developed an automatic for in vitro selection by assembling an affinity chromato-column, a PCR thermal cycler, a HPLC and a sample operation system. Several molecular biology methods were optimized for the machine. Automated selection was used to generate nucleic acid aptamers interacting specifically with an environmental contaminant. PMID- 12903348 TI - Accumulation and removal of heavy metal ions by insolubilized-DNA and its interaction. AB - DNA was immobilized onto a porous glass bead by a treatment with UV irradiation. The immobilized DNA was insoluble in water and used for accumulation of heavy metal ion. When DNA-immobilized glass bead was added into aqueous solution containing heavy metal ions, such as Hg2+, Cd2+, Pb2+, Zn2+, Cu2+ and Fe3+, the concentration of these metal ions in the solution was decreased. However, the concentration of Mg2+ in the solution was not affected by the addition of the DNA immobilized glass bead. These results suggested that UV-irradiated DNA selectively accumulated heavy metal ions. PMID- 12903349 TI - Control of DNA photocleavage by oligothiazole derivatives. AB - Mechanism of DNA damage by the p-cyanobenzamide (pCyBz)-oligothiazole derivatives were investigated by HPLC analysis. The oxidation product of dG by the photocleaver was imidazolone (dIz). PMID- 12903350 TI - Non-enzymatic oligomerization of racemic adenosine 5'-phosphorimidazolide on Na(+)-montmorillonite. AB - In this study, we have investigated non-enzymatic oligomerization of an activated racemic mononucleotide in the presence of Na(+)-montmorillonite. Oligomers up to the decamer in length were formed by oligomerization reactions of activated D- and L-mononucleotides. Similarly, oligomerization of an activated racemic mononucleotide results in the formation of oligomers up to the octamer. These results suggest that montmorillonite catalysis is quite efficient for the oligomerization of racemic monomers, though it is somewhat less efficient than that of D- and L-monomers. PMID- 12903352 TI - Preparation of DNA catenanes and observation of their topological structures by atomic force microscopy. AB - DNA catenanes have been prepared by the reaction of T4 DNA ligase with linear DNA in the presence of nicked DNA. Single molecular images of DNA catenanes and large circular DNAs have been clearly observed by AFM using a tapping mode at room temperature and in an ambient atmosphere. PMID- 12903351 TI - Classification of 3D structural character of RNA by hydrogen bond and base stacking. AB - We are developing a computational system to classify RNA structures by its structural character. Here, an improved grouping algorithm was introduced to the system and the base-stacking pattern (BSP) is used as a criterion for the classification in addition to hydrogen-bond pattern (HBP). 279 conformers of 15 mer RNA hairpin were classified into 89 and 36 groups by HBP and BSP, respectively, suggesting that HBP represents conformational character better than BSP. PMID- 12903354 TI - Structure and photodimerizations of 1-alkylthymines in single crystals. AB - 1-Alkylthymine gave four kinds of crystals depending on the solvents used. Photodimerizations of 1-alkylthymine in single crystals were found to depend on the crystal structure. In inactive crystals, the terminal methyl group of alkyl chains approached to the double bond of thymine, and inhibited rotation of thymine bases during photodimerization. PMID- 12903355 TI - Solution structure of the DNA complex with a quinacrine-netropsin hybrid molecule by NMR spectroscopy. AB - The solution structures of 1:1 complexes of a quinacrine-netropsin hybrid molecule with the self-complementary DNA duplexes, d(CGCGAATTCGCG)2 and d(CGAATTCG)2, have been studied by one- and two-dimensional 1H NMR spectroscopy. The NOE data indicate that the acridine ring of the hybrid intercalates into the 5'-GpA step and its netropsin moiety spans the minor groove of the central AATT region. PMID- 12903353 TI - Hydrolysis of DNA by Ce(IV)/EDTA complexes--the mechanism of DNA cleavage and design of artificial restriction enzymes. AB - Homogeneous Ce(IV) complex of EDTA promptly hydrolyzes oligonucleotides under physiological conditions. Moreover, the activity of Ce(IV)/EDTA for DNA hydrolysis is promoted by the addition of amines. When [Ce(IV)/EDTA] = 5 mumol dm3 and [ethylenediamine] = 100 mmol dm3, the catalytic activity is about 50 times as large as that of Ce(IV)/EDTA. The combination of Ce(IV)/EDTA and amines is eminent tools for the future molecular biology and biotechnology. PMID- 12903356 TI - Evaluation of the hydrogen bond energy of base pairs formed between substituted 9 methyladenine derivatives and 1-methyluracil by use of molecular orbital theory. AB - Systematic substituent effects on the stability of the hydrogen bonding between substituted 9-methyladenine derivatives (Ax) and 1-methyluracil (U) were studied by ab initio molecular orbital theory. Predicted substituent effects on the hydrogen bond energies of Ax-U base pairs were in good agreement with those observed for experimental binding constants. Ab initio calculation is effective for evaluation of the stability of the hydrogen-bonding pairs of chemically modified nucleic acid base analogues. In contrast to the substitution effect of uracil on hydrogen bond energies of A-Ux base pairs, it is difficult to systematically interpret the substitution effect of adenine derivatives for Ax-U base pairs. PMID- 12903357 TI - X-ray analyses of two DNA dodecamers containing N4-methoxy-cytosine paired with adenine or guanine. AB - To investigate mismatch of base-pairings in relation to mutagenesis by oxyamines, crystal structures of two DNA dodecamers with the sequence d(CGCZAA TTmo4CGCG) (Z = A or G), containing N4-methoxy-cytosine (mo4C), have been determined by X-ray analysis. These dodecamers essentially form right-handed B-form duplexes, respectively. In the dodecamer with Z = A, the two mo4C residues are adapted in imino form with the anti methoxyl group to form pairs with A on the opposite strand in a manner of Watson-Crick fashion. While in the dodecamer with Z = G, one mo4C in amino form with the anti methoxyl group forms a normal Watson-Crick pair with G, but the other one in imino form with syn methoxyl conformation wobbles with G. Based on these results, possible mutation mechanism has been proposed. PMID- 12903358 TI - Promotion of triplex formation by a fixed N-form sugar puckering: thermodynamic and kinetic studies. AB - We analyzed the effect of a fixed N-form sugar puckering of TFO (triplex-forming oligonucleotide) on the pyrimidine motif triplex formation at neutral pH, a condition where pyrimidine motif triplexes are unstable. Both thermodynamic and kinetic analyses revealed that the binding constant of the pyrimidine motif triplex formation at pH 6.8 with modified TFO containing the fixed N-form sugar puckering was about 20-times larger than that observed with unmodified TFO. Kinetic data also demonstrated that the observed increase in the binding constant at neutral pH by the fixed N-form sugar puckering resulted from the considerable decrease in the dissociation rate constant. Our results certainly support the idea that the fixed N-form sugar puckering of TFO could be a key modification and may eventually lead to progress in therapeutic applications of the antigene strategy in vivo. PMID- 12903359 TI - Quantitative detection of binding of PCNA protein to DNA strands on a 27 MHz quartz-crystal microbalance. AB - We report here binding kinetics of proliferating cell nuclear antigen (PCNA) to the dsDNA immobilized on a 27 MHz quartz-crystal microbalance. PCNA is a toroidal shaped protein and encircles dsDNA sliding along the strand. The binding behavior of this sliding clamp protein was compared with those of other side binding proteins such as GCN4-bZIP and Zn-finger peptides. PMID- 12903361 TI - Single long DNA molecule analysis using fluorescence microscopy. AB - Single long DNA molecule (T4 DNA) in agarose gel was visualized with a fluorescence microscope. We confirmed alternating current electric fields is effective for stretching of single DNA molecule in agarose gel. This stretching phenomenon was observed with wide range of agarose gel concentration from 0.5%(W/V) to 1.5%. From this observation, the presence of agarose gel fiber is essential for this stretching phenomenon. The stretching process of several DNA molecules in gel shows discontinuity, which is never observed in polymer systems. It would be based on topological restriction from gel fibers. PMID- 12903360 TI - In vitro selection by using mutated GCN4-bZIP peptides for analysis of peptide DNA interactions. AB - In vitro selection has been used as a method to determine the optimal binding site for DNA-binding proteins. We report here in vitro selection of dsDNA sequences that bind to mutated-GCN4-bZIP peptides. The GCN4-bZIP peptide mutated from alanine to histidine on a position-14 that contacts with DNA bound to different sequence from a binding site of wild type peptide. PMID- 12903362 TI - Translation activity of mitochondrial tRNA with unusual secondary structure. AB - Mammalian mitochondrial translation system requires two serine isoacceptor tRNAs, tRNA(Ser)GCU and tRNA(Ser)UGA, which correspond to codons AGY (Y = U or C) and UCN (N = A, G, C or U), respectively. Each tRNA has unusual secondary structure; tRNA(Ser)GCU lacks the entire D arm, while tRNA(Ser)UGA has a slightly varied cloverleaf structure with an extended anticodon stem. The translation activity of these tRNAs(Ser) was examined by using their transcripts produced by T7 RNA polymerase and an in vitro mitochondrial translation system, showing that tRNA(Ser)GCU has much lower translation ability than that of tRNA(Ser)UGA. HPLC analysis of the product suggested that a certain rate limiting step should exist in the elongation cycle of translation with tRNA(Ser)GCU. PMID- 12903363 TI - Trial for peptide bond formation using model molecules based on the interactions between the CCA sequence of tRNA and 23S rRNA. AB - The peptidyl transfer reaction catalyzed by the ribosome is a sophisticated product of evolution. The molecular mechanism of peptide bond formation has not been fully elucidated although the essential involvement of 23S rRNA has been established. The universal CCA sequence at the 3'-end of tRNA plays an important role in this process, by interacting with specific nucleotides in 23S rRNA. However, reconstitution of peptidyl transferase activity by a naked 23S rRNA (without the help of any of the ribosomal proteins) has not been reported. To investigate the possible evolutionary development of the peptidyl transfer reaction, we tried to obtain peptide bond formation using a piece of tRNA--an aminoacyl-minihelix--mixed with sequence-specific oligonucleotides that contained puromycin. This system reproduced conceptually the equivalent interactions between the CCA trinucleotide of tRNA and 23S rRNA. Peptide bond formation was detected by gel electrophoresis, TLC and mass spectrometry. These results have implications for the evolution of the peptidyl transfer reaction in biological system. PMID- 12903364 TI - Analysis of aptamer binding site for HCV-NS3 protease by alanine scanning mutagenesis. AB - Nonstructural protein 3 (NS3) of Hepatitis C virus (HCV) is a multifunctional protein and possesses protease, nucleotide triphosphatase and helicase activities. The N-terminal domain of NS3 (amino acids 1027-1218; delta NS3) has a trypsin-like protease activity and is essential for processing of viral polyprotein. Accordingly it is a potential target for anti-HCV drugs and we isolated RNA aptamers (Kd = 10 nM, Ki = 100 nM) using in vitro selection strategy. To study the interaction between delta NS3 and its aptamer, we applied alanine scanning mutagenesis and constructed seven mutant proteins at positive amino acid residues on the surface of delta NS3. Binding and inhibitory activities of the NS3 aptamer against mutant proteins were kinetically analyzed. These results clarified that especially Arg161 and Arg130 are important for interaction with the NS3 aptamer. PMID- 12903365 TI - Preparation of insolubilized-DNA film with three-dimensional network and removal of endocrine disruptors. AB - A water-insolubilized film was prepared by UV irradiation on a dried DNA film. When a UV-irradiated DNA was examined using a circular dichroism spectroscopy, a double stranded structure was observed as well as that of native DNA. The UV irradiated DNA film was also accumulated intercalating reagents. These results suggested that the double stranded structure was involved in the UV irradiated DNA film with a three-dimensional network. The thymine-thymine dimer formation was suggested to be involved in the cross-linking reactions by the polymerization analysis using poly(dA)-poly(dT) and poly(dG)-poly(dC). We also demonstrate the utilization of the UV-irradiated DNA film as a functional material for the accumulation of harmful DNA intercalating pollutants in aqueous solution. These results suggested that the UV-irradiated DNA film was applicable as a functional material for medical, engineering and environmental sciences. PMID- 12903366 TI - Automated, solid-phase coupling of rhodamine dye acids to 5' amino oligonucleotides. AB - A convenient method has been developed for directly labeling the 5' amino group of oligonucleotides on solid-support with rhodamine dyes. PMID- 12903367 TI - Histone acetyltransferase (HAT) activity of ATF-2 is necessary for the CRE dependent transcription. AB - ATF-2 is a DNA-binding protein that binds to cAMP-response elements (CREs) and forms a hetrodimer with c-Jun, via binding of the leucine zipper motif and then stimulates the CRE-dependent transcription (1,2). Recently, we have reported that ATF-2 has an intrinsic acetyltransferase activity that is controlled by phosphorylation. Mutant form of either p300 or ATF-2 with mutations in the HAT domain failed to stimulate the CRE-dependent transcription in response to UV irradiation. Moreover, phosphorylation of ATF-2 enhanced its HAT activity and CRE dependent transcription. Thus, these results indicate that HAT activity of ATF-2 is important for the CRE-dependent transcription. PMID- 12903368 TI - Unnatural base pairs between 2-amino-6-(2-thienyl)purine and the complementary bases. AB - The unnatural base, 2-amino-6-(2-thienyl)purine (designated as s), instead of 2 amino-6-(N,N-dimethylamino)purine (designated as x), was designed in order to improve the specificity and efficiency of the base pairing with pyridin-2-one (designated as y). DNA fragments containing s were chemically synthesized, and the thermal stability and the enzymatic reactions involving the s-y pairing were examined. Thermal denaturation experiments showed that the DNA duplex (12-mer) containing the s-y pair was more stable than that containing the x-y pair. The incorporation of dyTP was also more advantageous to the s-y pairing than the x-y pairing in single-nucleotide insertion experiments using the Klenow fragment of Escherichia coli DNA polymerase I. PMID- 12903369 TI - The structure and function of tRNA-like domain in E. coli tmRNA. AB - tmRNA has a dual function both as a tRNA and as an mRNA in trans-translation. tmRNA possesses structural elements similar to canonical tRNAs. Our mutation studies show that the tRNA-like structure is crucial for function as an mRNA as well as a tRNA. PMID- 12903370 TI - Zn finger and nuclear localization of transcription factor Sp1. AB - Transcription factor Sp1 is located in the nucleus of a mammalian cell and importantly related to expression of many cellular genes. In order to elucidate the nuclear localization mechanism of Sp1, various truncated fragments of Sp1 were fused to green fluorescent protein (GFP) and expressed in HeLa cell. The results show significance of the DNA binding region, especially, zinc finger (Zn finger) domain for nuclear localization of Sp1 in HeLa cell. PMID- 12903371 TI - Posttranscriptional modification of transfer RNA in the submarine hyperthermophile Pyrolobus fumarii. AB - In the RNA of hyperthermophiles, which grow optimally between 80 degrees C and 106 degrees C, posttranscriptional modification has been identified as a leading mechanism of structural stabilization. Particularly in the Archaeal evolutionary domain these modifications are expressed as a structurally diverse array of modification motifs, many of which include ribose methylation. Using mass spectrometric techniques we have examined the posttranscriptional modifications in unfractionated tRNA from the remarkable organism Pyrolobus fumarii, which grows optimally at 106 degrees C, but up to 113 degrees C (Blochl et al. (1997), Extremophiles, 1, 14-21). Twenty-six modified nucleosides were detected, 11 of which are methylated in ribose. A new RNA nucleoside, 1,2'-O-dimethylguanosine (m1Gm) was characterized and the structure confirmed by chemical synthesis. PMID- 12903372 TI - Screening in vivo for RNA-binding peptides from combinatorial libraries. AB - We have modified a previously developed genetic assay system for RNA-polypeptide interactions in a attempt to more readily identify RNA-binding peptides. The first modification involved the design of a "complex" library that would contain a variety of RNA-binding polypeptides. The second modification involved the use of neomycin phosphotransferase (NPT II) as the reporter gene, therefore allowing "selection" of RNA-binding peptides by kanamycin resistance. The improved screening system should allow the identification of peptides that bind to a variety of RNA structures. PMID- 12903373 TI - Affinity photo-crosslinking study of the DNA base flipping pathway by HhaI methyltransferase. AB - HhaI DNA methyltransferase flips the inner cytosine in the recognition sequence 5'-GCGC-3' out of the DNA helix and into the catalytic site for methylation. To identify intermediate states on the base-flipping pathway, affinity photo crosslinking experiments were performed with synthetic dodecamer duplexes containing modified bases 2-thiothymine (2sT) or 4-thiothymine (4sT) at the target base position. Here we show that the DNA strand containing 2sT, but not 4sT, covalently cross-links to the HhaI methyltransferase upon irradiation at 340 360 nm. PMID- 12903374 TI - NO induced novel DNA lesions: formation mechanism. AB - 2'-Deoxyguanosine was treated with NO/O2 mixture at pH 7.0-7.8, and as well as the known major products such as 2'-deoxyxanthosine and 2'-deoxyoxanosine, some unidentified products were detected by RP-HPLC. In the present study, one of them has been characterized as a novel lesion, N2-nitro-2'-deoxyguanosine by spectrometric and chromatographic analysis. The mechanism for the production is also discussed. PMID- 12903375 TI - Extension of biochemical functions by the introduction of nonnatural amino acids and artificial nucleic acid analogs. AB - Extension of biochemical functions has been attempted by introducing nonatural amino acids and artificial nucleic acid analogs. Nonnatural amino acids have been linked to tRNAs and the amino-acylated tRNAs were added to E. coli in vitro protein synthesizing system to produce nonnatural mutant proteins. The positions of the nonnatural amino acids have been assigned by the 4-base codons, like CGGG and AGGU. The extended codons have been introduced at a specific position or at random positions on a DNA. In the latter case, a DNA library that contains a single 4-base codon at random positions can be obtained. The combination of these new techniques opens a way to the introduction of artificial functions to biochemical systems. PMID- 12903376 TI - Photo-regulation of DNA/RNA duplex formation by azobenzene-tethered DNA towards antisense strategy. AB - Modified DNA carrying an azobenzene was successfully applied to the photo regulation of DNA/RNA hybridization. When the azobenzene was isomerized from trans- to cis-form on UV-irradiation, the melting temperature of the duplex was significantly lowered. This process was totally reversible so that the Tm increased by cis-->trans isomerization induced by visible light irradiation. PMID- 12903377 TI - Two-terpyridine.Cu(II) complexes-containing antisense systems for rapid and highly site-specific RNA cleavage. AB - In an approach toward artificial ribonucleases, novel RNA cleaving systems were constructed that contained two terpyridine.Cu(II) residues. The first antisense system used tandem Cu(II) complex--2'-O-methyloligonucleotide 5'- and 3' conjugates to cleave an RNA substrate. The second system, which will be described in a future paper, contained two contiguous Cu(II) complex residues at an internal site of a 2'-O-methyloligonucleotide. We found that the first system rapidly cleaved RNA with high site-specificity. Based on these results, we expect the second system to also show efficient RNA cleavage. PMID- 12903378 TI - NMR solution structure of the L 9.1a region of Tetrahymena group I intron. AB - The solution structure of 20 mer RNA contained of the loop 9.1a region of Tetrahymena group I intron was studied by NMR. This RNA oligomer has hairpin and duplex structures at high concentration (1 mM) of the sample even at low NaCl concentration (5 mM). In the hairpin structure, GC base pairs by the loop-loop interaction are formed. As study of NOESY measurements, and by the compared with the sequence, this loop region is presumed to interact with the loop 5c. PMID- 12903379 TI - Suppression of BCR-ABL mRNA by various ribozymes in HeLa cells. AB - Ribozymes are RNA molecules with enzymatic activity that can cleave target RNA molecules in a sequence specific manner. To date, various types of ribozyme have been constructed to cleave other RNAs and such trans-acting ribozymes include hammerhead, hairpin and HDV ribozymes. External guide sequence (EGS) can also induce the suppression of a gene-expression by taking advantage of cellular RNase P. Here we compared the activities of various functional RNA cleavers both in vitro and in vivo. The first purpose of this comparison was intended to determine the best ribozyme motif with the highest activity in cells. The second purpose is to know the correlation between the activities of ribozymes in vitro and in vivo. Our results indicated that the intrinsic cleavage activity of ribozymes is not the sole determinant that is responsible for the activity of a ribozyme in cultured cells. PMID- 12903380 TI - NMR analysis of tertiary interactions in HDV ribozymes. AB - Three variants of minimized hepatitis delta virus (HDV) RNA ribozyme systems designed on the basis of the "pseudoknot" model were synthesized and their tertiary interactions were analyzed by NMR spectroscopy. Rz-1 is a cis-acting ribozyme system (the cleaved form, 56-mer) in which stem IV is deleted from the active domain of genomic HDV RNA. Rz-1 was uniformly labeled with stable isotopes, 13C and 15N. Rz-2 is a trans-acting ribozyme system (substrate: 8-mer, the cytidine residue at the cleavage site is replaced by 2'-O-methylcytidine; enzyme: 16-mer plus 35-mer). Rz-2 was partially labeled with stable isotopes in guanosine residues of enzyme 35mer. Rz-4 is a trans-acting ribozyme system (substrate: 8mer, the cytidine residue at the cleavage site is replaced by 2'-O methylcytidine; enzyme 53mer) which was designed by Perrotta and Been. Rz-4 has the same sequence and an extra loop closing stem IV. From 2D-NOESY and 2D-HSQC (except for Rz-4) spectra, it was suggested each ribozyme forms "pseudoknot" type structure in solution. Additionally, it was found that G38 of Rz-1, G28 and G29 of Rz-2 and Rz-4 form base-pairs. These novel base-pairs are observed in the crystal structure of a modified genomic HDV RNA. From temperature change experiment of Rz-2, the imino proton signal of G28 disappeared at 50 degrees C earlier than the other corresponding signals. Upon MgCl2 titration of Rz-2, this signal showed the largest shift. PMID- 12903381 TI - Inhibition of influenza virus RNA (PB2 mRNA) expression by a modified DNA enzyme. AB - DNA enzymes are RNA-cleaving single stranded DNA molecules. The structure and the catalytic domain of a DNA enzyme were determined by Santro et al. in 1997. In this study, we have designed several types of DNA enzymes (PB2Dz) targeted to the PB2 mRNA translation initiation region of influenza A virus, and examined their cleavage kinetics, nuclease resistance, and a luciferase gene reporter assay. Using a synthetic substrate, these DNA enzymes were shown to have cleavage activity that is dependent on the length of the substrate recognition domain. To confer serum nuclease resistance to the DNA enzymes, we designed a new type of DNA enzyme that has the N3'-P5' phosphoramidate modification (PB2Dz-N) at each terminal. We examined the activity of this DNA enzyme in vivo. The DNA enzymes used in this study inhibited the expression of the PB2-luciferase gene in COS cells. These results suggest that DNA enzymes are potentially useful as gene inactivating agents of influenza A virus. PMID- 12903382 TI - Acceleration of DNA strand exchange by polycation comb-type copolymer. AB - The accelerating effect of cationic substances on DNA strand exchange reaction between 20 bp DNA duplex and its complementary single strand was studied. A comb type polycationic copolymer which is composed of poly (L-lysine) backbone and dextran graft chain (PLL-g-Dex) and known to stabilize triplex DNA expedites the strand exchange reaction under physiological relevant conditions. Electrostatically small excess of the copolymer increased DNA strand exchange rate by 300-fold while large excess of spermine or cethyltrimethylammonium bromide, cationic detergent known to promote markedly hybridization of complementary DNA strands, showed slight effect. It should be noted that the copolymer promotes the strand exchange reaction while it stabilizes double stranded DNA. PMID- 12903383 TI - Fluorescent labelling of ribonucleosides at 2'-terminus; comparative fluorescence studies. AB - In case of RNA's, multiple labelling can be achieved by exploiting the available 2'-OH position of sugar moiety of nucleosides. N-protected nucleoside viz. cytidine has been prepared using a selective photolabile group i.e. 2 nitrobenzyloxycarbonyl. After protection of 5',3'-OH with 1,1,3,3,-tetraisopropyl disiloxyl group, 2'-OH was selectively activated by using N,N'-carbonyl diimidazole (CDI) and subsequently condensed with dansyl amide. After usual deprotection step comparative fluorescence studies of the monomer were carried out using different solvents/buffers. PMID- 12903384 TI - Salt-dependent folding of long duplex DNA by histone H1. AB - It is found that T4 phage DNA complexed with histone H1 assembled into a string of-bead structure, when the complex is prepared by a gentle diluting procedure from a high salt solution (2 M NaCl) to a low salt solution (50 mM NaCl). We used fluorescence microscopy to perform the real-time observation on formation and motion of a string-of-bead structure. Spatial histone H1 distribution on the DNA H1 complex is observed by immuno-fluorescence microscopy. PMID- 12903385 TI - The engineering, structure, and DNA binding properties of a novel His4-type zinc finger peptide. AB - We have created a novel His4-type zinc finger protein (H4Sp1) engineered by Cys- >His mutations of the Cys2His2-type zinc finger in transcription factor Sp1. The CD and NMR studies reveal that the His4 domain has Zn(II)-dependent folding properties and similar secondary structures to wild-type Cys2His2 domain. The DNA binding experiments demonstrate that H4Sp1 can bind DNA in a specific way. The present artificial peptide H4Sp1 will provide valuable information about the interaction between a metallopeptide and DNA. PMID- 12903386 TI - Smart probe: a novel fluorescence quenching-based oligonucleotide probe carrying a fluorophore and an intercalator. AB - A novel probe (Smart probe) has been developed for nucleic acid detection. The smart probe is an oligodeoxyribonucleotide carrying a fluorophore and an intercalator internally. Fluorescence of the smart probe is quenched by the intercalator in the absence of target sequence. While upon hybridization the probe emits greater fluorescence due to the interference of quenching by intercalation. The smart probe has been shown to recognize a single base mismatch in the double-stranded form without utilizing thermal stability difference of hybrids. PMID- 12903387 TI - Effect of serine residue on the effectiveness of cationic polypeptide-based gene delivery. AB - Poly-L-lysine(pL) was chemically modified based on two essential features which we recently reported and subjected to the gene-transfer experiment in vitro. Introduction of 25 mol% serine residue to pL slightly enhanced the gene expression level, while trimethylation of epsilon-ammonium groups of lysine did not. Only when pL was modified in both way, giving N2-trimethyl poly(lysine-co serine), markedly enhanced gene expression was observed. The cellular uptake and localization of DNA in the cells were similar for each cationic polypeptide. DNA forming complex with the polypeptides containing serine residue was found to be well transcribed in in vitro transcription/translation system, suggesting the hydrophilic nature may allow polypeptide/DNA complexes to be recognized by the transcriptional factors and lead the subsequent effective gene expression. PMID- 12903388 TI - [Multiple types of chemokines expressed in mouse thymic stromal cell lines]. AB - OBJECTIVE: To investigate semi-quantitative mRNA expression of SDF-1 alpha, IP 10, KC, MCP-1, and RANTES in thymic stromal cell lines of MTEC1, MTDC, D2SC, MTECB, TEC 1C8, TNC as well as in the primary thymic stromal cell cultures. The chemotactic activities of recombinant SDF-1 alpha, IP-10, MCP-1, and RANTES to mouse thymocytes were detected. METHODS: Using beta-actin as internal control, the mRNA of the chemokines listed above were amplified for 30 cycles with RT-PCR. The amplified products were observed by agarose electrophoresis, and each band was analyzed with integrated optical density. With the method of Boyden chamber assay, the chemotactic activities of recombinant SDF-1 alpha, IP-10, MCP-1, and RANTES were detected to thymocytes, and the chemotactic indices were calculated. RESULTS: The expression intensity of SDF-1 alpha, IP-10, KC, MCP-1, and RANTES varied from each other in the stromal cell lines detected. The PCR products of SDF-1 alpha and MCP-1 were not seen in D2SC or TEC 1C8, nor was the band of KC observed in TEC 1C8 either. The chemotactic indices of recombinant SDF-1 alpha, IP-10, MCP-1, and RANTES to thymocytes were 3.7, 4.5, 6.2, and 2.6, respectively. CONCLUSIONS: Different thymic stromal cell lines could express different types of chemokines with different expression intensities. To thymocytes, recombinant SDF 1 alpha, MCP-1, and IP-10 showed strong chemotactic activities, while the chemotactic activity of RANTES was very weak. PMID- 12903389 TI - [Preparation and identification of monoclonal antibodies against human apoptosis related protein TFAR19]. AB - OBJECTIVE: To obtain mouse anti-human TFAR19 monoclonal antibodies for further study of the structure and function of the apoptosis-related protein TFAR19. METHODS: BALB/c mice were immunized with recombinant human TFAR19, hybridoma cells were screened by cell fusion and subcloning approach. The monoclonal antibodies were identified by ELISA and Western blotting. RESULTS: Three hybridoma cells (C1, C10, 2C12) stable in secreting anti-TFAR19 monoclonal antibodies were obtained. The monoclonal antibodies showed high specificity and high titer to TFAR19 with various affinity. All of them belong to IgG1 subclass. CONCLUSIONS: These monoclonal antibodies could bind specifically to TFAR19 protein. PMID- 12903390 TI - [Alloreactive helper T cell responses in kidney transplant patients]. AB - OBJECTIVE: To study allo-specific immune responses of CD4+ helper T lymphocytes (Th) in patients with long-term and well-functioning renal grafts. METHODS: Peripheral blood mononuclear cells (PBMC) were collected from 16 patients with long-term (3-15 years) and well-functioning live-related renal grafts. Donor specific Th precursor (Th-p) frequencies in the PBMC were determined by limiting dilution analysis(LDA). RESULTS: Eight out of the 16 patients showed donor specific hyporesponsiveness. Donor-specific Th-p from the other 8 patients was susceptible to anti-CD4 inhibition. CONCLUSION: Some of the patients carrying long-term and well-functioning renal grafts had become immunologic tolerant to donor cells. Donor-specific Th-p from patients who did not show donor-specific hyporesponsiveness were of low avidity type. PMID- 12903391 TI - [Induction of skin immune tolerance of mouse to rat xenogeneic transplantation]. AB - OBJECTIVE: To explore a moderate and effective project which will be clinically suitable to induce donor-specific tolerance across xenogeneic barriers. METHODS: 4 x 10(7) Lewis rat bone marrow cells were infused to 300 rad total body irradiated (TBI) C57BL/6 (B6) mice, combined with intraperitoneal administration of cyclophosphamide (CTX) and intravenous injection of anti-mouse CD4 monoclonal antibody. Recipient B6 mice were characterized for the tolerance status with donor Lewis rat skin graft, mixed lymphocyte reaction (MLR), and delayed-type hypersensitivity (DTH) assays after 30 days. Adoptive transfer and the effect of IL-2 on MLR were detected to further explore the tolerance mechanism. RESULTS: Survival time of donor rat skin grafts was specifically prolonged in the tolerant B6 mice. The results of MLR and DTH assays showed donor-specific hyporeactivity, while the tolerant B6 mice were still immunocompetent to MHC-disparate third party BALB/c mouse or DA rat stimulator cells. CONCLUSIONS: A reliable xenogeneic transplantation tolerance of mouse to rat was achieved by this project of tolerance induction. The result of in vitro and in vivo adoptive transfer of spleen cells from tolerant B6 mice demonstrated that the suppressor cells were unlikely to exert effect in the tolerance. The inhibition of specific MLR could be reversed by adding IL-2, indicating that clonal anergy instead of clonal deletion is responsible for the tolerant maintenance. PMID- 12903392 TI - [Construction of BPI23-Fc gamma 1 recombinant protein prokaryotic expression vector and the expression and biological assessment of BPI23-Fc gamma 1 recombinant protein]. AB - OBJECTIVE: To construct pBV-BPI600-Fc gamma 1(700) recombinant expression vector, to transform into Escherichia coli DH5 alpha, and to induce the expression of BPI23-Fc gamma 1 anti-bacterial recombinant protein. METHODS: Genes which encode BPI23 and Fc gamma 1 were amplified by RT-PCR from mRNA that was extracted from HL-60 cell and normal human leukocytes; Recombinant cloning vector and recombinant expression vector were constructed. pBV-BPI600-Fc gamma 1(700) recombinant expression vector was transformed into the competent Escherichia coli DH5 alpha and BPI23-Fc gamma 1 recombinant protein was expressed by temperature induced method. RESULTS: (1) Expected amplified products BPI600 bp and Fc gamma 1(700) bp were obtained by RT-PCR. (2) pUC18-BPI180, pUC18-BPI420, and pUC18-Fc gamma 1(700) recombinant cloning vector were successfully constructed, and sequences were identical with the reported ones. (3) pBV-BPI600-Fc gamma 1(700) recombinant expression vector was successfully constructed, and results of the enzyme digestion analysis were identical with expected ones. (4) pBV-BPI600-Fc gamma 1(700) recombinant expression vector was transformed into the competent Escherichia coli DH5 alpha and BPI23-Fc gamma 1 recombinant protein was expressed by temperature-induced method, and its expression level was accounted for 20% of total bacterial proteins. (5) The renatured BPI23-Fc gamma 1 recombinant protein had anti-bacterial activity and biological functions of complement fixation, opsonization. CONCLUSION: pBV-BPI600-Fc gamma 1(700) recombinant expression vector was successfully constructed, and BPI23-Fc gamma 1 recombinant protein with BPI and IgGFc double biological activity was expressed in Escherichia coli. PMID- 12903393 TI - [Expression of the gene encoding human leukocyte differentiation antigen 5D4]. AB - OBJECTIVE: To make sure that 5D4 cDNA is a full length cDNA and to prepare antigens to make monoclonal antibodies against different epitopes in 5D4 protein. METHODS: Recombinant expression vectors pGEX-5X-3-5D4 (full length) and pGEX-5X-3 5D4 (5'fragment) were constructed. 5D4 polypeptides were expressed in Escherichia coli DH5 alpha. 5D4 mRNA expression was detected in various tissues and cell lines with Northern blot assay and RT-PCR. RESULTS: Northern blot demonstrated five mRNA transcripts of 1.9, 2.4, 3.9, 4.5, and 10.0 kb in length existing and expressing of various extent in various tissues. The 1.9 kb mRNA transcript was observed only in the spleen and the small intestinal tissues. The expression of 5D4 mRNA was different in various cell lines. 5D4 polypeptides were expressed in Escherichia coli DH5 alpha. CONCLUSION: The 1,846 bp 5D4 cDNA is a full length cDNA. A full length 5D4 protein and a 5D4 polypeptide which is the NH2 side to the transmembrane region fused with GST with a molecular weight of 66,000 and 60,000 expressing in Escherichia coli DH5 alpha. PMID- 12903394 TI - [Identification of functional sites of human IgA Fc receptor (CD89)]. AB - OBJECTIVE: To identify the domains and residues in IgA Fc receptor (FC alpha R) essential for interaction with IgA. METHODS: Various fragments of Fc alpha R cDNA were amplified by PCR or site-directed mutagenesis PCR. The amplified products were ligated into the EcoR I and Xho I sites of the pET30a plasmid, a prokaryotic expressing vector. Then the recombinant plasmids were transformed into Escherichia coli BL21. Plasmids with correct cDNA fragment inserts were identified by digestion with two restriction enzymes and by DNA sequencing. Expression of cDNA fragments was made by isopropyl-beta-D-thiogalactopyranoside (IPTG) induction. Western blotting was carried out to identify the functional site of Fc alpha R. Monoclonal antibodies against Fc alpha R were used as primary antibodies. RESULTS: Five recombinant plasmids expressing various fragments of Fc alpha R were constructed. The monoclonal antibodies that can blot the IgA binding sites of Fc alpha R could only bind to the membrane-distal domains of Fc alpha R (EC1). CONCLUSION: The functional site of Fc alpha R was located at EC1 domain. PMID- 12903395 TI - [Areas of brain involved in immunoregulation]. AB - OBJECTIVE: To study the location of brain areas involved in immunoregulation. METHODS: Immunohistochemistry and immunofluorescence were used to detect the different distribution of cytokines immunopositive cells in the brain of rats immunized via both intraperitoneal and subcutaneous injections. RESULTS: The cytokines immunopositive cells were distributed in the supraoptic (SO) and paraventricular (PV) nuclei of the hypothalamus, the anterior hypothalamic (AH) nucleus, arcuate and median eminence, the lateral hypothalamic nucleus (LH) and the amygdaloid nuclear complex while only the appearance of the cytokines immunopositive cells in LH and amygdaloid nuclear complex in hypothalamus was related with immunization status of the animals. Double-labelling results showed that the cytokines immunopositive cells were neurons. CONCLUSIONS: We have observed that neurons of the LH and amygdaloid nuclear complex in hypothalamus as a main source of the neuroimmunoregulation played a key role in neuroimmunoregulation and they participated in the neuroimmunoregulation at an early stage of the immune response. PMID- 12903397 TI - [Expression and kinase activity of B-Raf and Raf-1 in leukemia cell lines]. AB - OBJECTIVE: To study expression and kinase activity of B-Raf and Raf-1 kinases in several leukemia cells and normal peripheral blood lymphocytes. METHODS: Western blot was carried out to detect expression of B-Raf and Raf-1 in peripheral blood lymphocytes and leukemia cells. Kinase activities of Raf-1 and B-Raf were analyzed by immunoprecipitation and Western blot. RESULTS: The expression levels of Raf-1 were similar in tested leukemia cells and normal peripheral blood lymphocytes, and Raf-1 protein was all minimally tyrosine-phosphorylated. B-Raf expressed in all tested leukemia cells, but not in normal peripheral blood lymphocytes, and B-Raf was highly activated in Jurkat and K562 leukemia cells. CONCLUSION: The abnormal expression and kinase activity of B-Raf in leukemia cells might be related to leukemia etiologically. PMID- 12903396 TI - [Effect of 6A8 alpha-mannosidase expression on the proliferative response of human B cell line 3D5]. AB - OBJECTIVE: To study the effect of 6A8 alpha-mannosidase expression on the proliferative response of human B cell 3D5. METHODS: Recombination of adeno associated virus vector(rAAV) mediated delivery of sense 6A8 DNA or antisense 6A8 DNA into 3D5 cells, and monoclonal antibody 6A8 alpha staining and Con A binding assay for determination of the change of 6A8 alpha-mannosidase expression, MTT assay for proliferation detection of 3D5 cells driven by Staphylococcus aureus crude cell suspension formalin-fixed (SAC), lower molecular weight B cell growth factor (LMW-BCGF), or rIL-6. RESULTS: The expression of 6A8 alpha-mannosidase was enhanced in sense 6A8-transduced cells and reduced in antisense 6A8-transduced cells. In comparison with the wild type and the mock-transduced cells, the proliferative response of the sense 6A8-transduced cells to SAC stimulation was enhanced (P < 0.05). However, transduction with antisense 6A8 did not affect the response. In addition, transduction with either sense or antisense 6A8 had no effect on proliferation of 3D5 cells induced by LMW-BCGF or IL-6. CONCLUSION: The proliferative response to SAC stimulation was enhanced in the 3D5 cells with enhanced expression of 6A8 alpha-mannosidase. Either enhancement or reduction of 6A8 alpha-mannosidase expression had no effect on proliferation induced by LMW BCGF or IL-6. PMID- 12903398 TI - [Interactions between domains within the NH2- and COOH-terminal fragments of presenilins]. AB - OBJECTIVE: To analyze the interactions between domains within the NH2- and COOH terminal regions of presenilins. METHODS: The various constructions corresponding to NH2-terminal fragment (NTF) and COOH-terminal fragment (CTF) derivatives of presenilin 1 (PS1) and presenilin 2 (PS2) were generated by RT-PCR, and their interactions were assayed by yeast two-hybrid system. RESULTS: Domains within the NH- and COOH-terminal fragments of presenilins could directly interact with each other, and therefore form high molecular weight complex. The interaction site between domains within PS1 located at amino acid 361-447 of PS1 CTF, without the involvement of other partners. Similar interaction was not observed between PS11 360 and PS2341-448, PS2(1)-340 and PS1(361)-467. CONCLUSIONS: Intramolecular interaction between domains within the NH2- and COOH-terminal regions of presenilins may be critical to the folding and assembly of mature PS molecules. PMID- 12903399 TI - [Effect of reperfusion after local ischaemia on the M receptor density in rat brains]. AB - OBJECTIVE: To study the dynamic changes in the density of muscarinic acetylcholine receptor in different areas of rat brains during reperfusion after local ischaemia. METHODS: The rat cerebral focal ischemia/reperfusion model after middle cerebral artery occlusion was made by the intraluminal suture method. Using 3H-QNB as radioligand, after binding with M receptor on the brain slice of the rat model, the brain samples were exposed on the hypersensitive film [H-3] to complete autoradiography, and the density of M receptor in different areas on the film of autoradiography was measured. RESULTS: There were no changes in the M receptor density in 2 hours' ischaemia group and 2 hours' reperfusion group, M receptor densities in the area of occipital cortex (OC) and temporal cortex (TC) were observed to decrease significantly in 8 hours' reperfusion group, but the degree of decrease was lower than 24 hours' and 72 hours' reperfusion groups, there was significant decrease in the density of M receptor in the areas of caudate putamen (CP), frontal cortex (FC), TC, OC of the 24 hours' and 72 hours' reperfusion groups and no difference was found between these two groups, and the imaging of M receptor in the domains with decreased receptor was obvious. CONCLUSIONS: The quantity of M receptor did not change right after ischaemia or reperfusion. It decreased gradually after an interval of time; Imaging of the M receptor could assess the changes of the density of viable cholinergic neurons and be a marker of diagnosis, therapy, and prognosis of brain ischaemia. PMID- 12903400 TI - [Effects of 1 alpha(OH)D3 on trabecular and cortical bone in ovariectomized rats]. AB - OBJECTIVE: To study the pharmaceutical effects of 1 alpha(OH)D3 on trabecular and cortical bone in ovariectomized (OVX) rats. METHODS: 41 female Wistar rats of six months old were randomly divided into 5 groups: (1) Baseline control; (2) Sham control; (3) 6 weeks after OVX (OVXb); (4) 14 weeks after OVX (OVXe); (5) OVX + 0.1 microgram/(kg.d) 1 alpha(OH)D3 (O + VD), treatment started 6 weeks after OVX and lasted 8 weeks. Histomorphometry analysis of tibia, peripheral quantitative computed tomography (pQCT) scanning of femur, bone biomechanical test and serum and urinary biochemical parameters were determined. RESULTS: The levels of bone turnover indexes increased in OVX rats, OVX also resulted in reducing of trabecular, bone mass and biomechanical properties. The ratio of urinary deoxypyridinoline crosslink/creatinine was decreased by 67.0% in O + VD group compared with OVXe group 67.0% [(43.50 +/- 11.20) nmol.L-1/mmol.L-1 vs(131.80 +/- 14.90) nmol.L-1/mmol.L-1, P < 0.01]. Percent trabecular area (Cn-BV/TV) was increased by 89.8% in O + VD group compared with OVXe group (11.03 +/- 0.73 vs 5.81 +/- 1.29, P < 0.05). Trabecular bone mineral content and density were increased by 77.3% and 91.3% compared with OVXe respectively (P < 0.05). Although cancellous maximal load and stiffness increased in O + VD group, but no statistical significance. 1 alpha(OH)D3 also enhanced polar moment of inertia (PMI) and maximal load of cortical bone in femur compared with OVXe (23.70 +/- 1.63 vs 18.23 +/- 1.41, P < 0.01 and 171.69 +/- 9.92 vs 147.58 +/- 11.29, P < 0.05 respectively). CONCLUSION: 1 alpha(OH)D3 inhibited the higher bone turnover induced by OVX, increased trabecular bone in proximal tibia and bone mass in distal femur. 1 alpha(OH)D3 also improved the mechanical properties of cortical bone in femur. PMID- 12903401 TI - [Surgical treatment of malignant esophageal tumors in PUMC Hospital]. AB - OBJECTIVE: To study how to prolong the postoperative survival time of the patients with malignant esophageal tumors. METHODS: The clinical data of 1,098 patients with malignant esophageal tumors from 1961 to 1992 were retrospectively analyzed. The deletion of fragile histamine triplet(FHIT) gene(a tumor suppressor gene) in 30 fresh esophageal samples obtained in 1996 was detected with PCR and RT-PCR. RESULTS: The resectability was raised gradually and the operative morbidity and mortality decreased year-by-year, but there was no significant improvement on the postoperative 5-year survival rate. Delayed diagnosis and ir radical resection influenced the long term survival. The deletion of cDNA of FHIT gene was 64.2% in esophageal cancer and 20% in the resected margin of the cancer. CONCLUSIONS: High atypical hyperplasia in esophageal epithelium and deletion of FHIT gene in esophageal cancer and its resected margin are pathological and molecular markers for early diagnosis of esophageal cancer respectively, and the latter may be one of the molecular markers for the resection. Early diagnosis and treatment, radical resection, and postoperative nutritional support are essential for the improvement of the postoperative survival time of the patients. PMID- 12903402 TI - [Survival motor neuron gene and neuronal apoptosis inhibitory protein gene deletion in patients with spinal muscular atrophy]. AB - OBJECTIVE: To investigate the frequencies of gene deletion survival motor neuron telomere (SMNTel) exon 7 and neuronal apoptosis inhibitory protein gene (NAIP) exon 5 in 55 Chinese spinal muscular atrophy (SMA) patients, and compare the relationship between these two candidate genes and the disease. METHODS: PCR-SSCP method was used to detect the deletion of SMNTel exon 7, direct visualization of PCR products by agarose electrophoresis was used to detect the deletion of NAIP exon 5 in 55 SMA patients with type I and type II. 40 normal individuals were involved in the study as controls. RESULTS: Homozygous deletion of the SMNTel exon 7 was identified in 92% (23/25) of SMA type I patients and 90% (27/30) of SMA type II patients. The same deletion was found in two mothers and one father of SMA patients. There was no homozygous deletion found in normal controls. None of the homozygous deletion of NAIP exon 5 was found in 55 SMA patients and normal controls. Only two patients were found to have the heterozygous deletion. CONCLUSIONS: The frequency of homozygous deletion of SMNTel exon 7 was 90.1%. Our data support that SMN gene is strongly associated with SMA. PMID- 12903403 TI - [Expression of epidermal growth factor receptor in gestational trophoblastic tumours]. AB - OBJECTIVE: To investigate the expression of epidermal growth factor receptor (EGFR) in gestational trophoblastic tumours and its clinical significance. METHODS: Placentas of 10 normal early pregnancies, 20 molar pregnancies, 10 samples from invasive mole and 8 samples of choriocarcinoma were used for the study. EGFR expression was evaluated by immunohistochemistry using monoclonal antibody. RESULTS: EGFR was expressed both in normal chorionic villi and tissues of trophoblastic tumours. EGFR was expressed more strongly in normal chorionic villi and non-malignant transformed hydatidiform mole than in malignant transformed molar pregnancies and malignant trophoblastic tumours (P < 0.05). Univariate and multivariate analysis indicated that EGFR immunostaining intensity was significantly related to the clinical stage in malignant trophoblastic tumours (P = 0.026, OR = 9.874). CONCLUSION: EGFR expression could be used as an index for prediction of malignant transformation of hydatidiform mole and staging of malignant trophoblastic tumours. PMID- 12903404 TI - [Immobilization and properties of angiotensin converting enzyme]. AB - OBJECTIVE: To study the immobilization of angiotensin converting enzyme (ACE) for purifying ACE inhibitor from a native peptide mixture. METHODS: The experiment was carried out under the low water activity condition, using tosylate chloride activating side-chain hydroxyl group of Sepharose CL-4B agarose to form a high active group which could react with the free amino-group of ACE to link the enzyme with agarose. RESULTS: Immobilized ACE not only had a wider pH range, but also had a 0.6 unit right-move optimum pH than soluble ACE. After treated in pH9.0 and pH 6.5 conditions for 24 h respectively, 82% and 68% enzyme activities of immobilized ACE was maintained, and soluble ACE remained 64% and 39%. Immobilized and soluble ACE both appeared maximum enzyme activity at about 50 degrees C, the soluble ACE would lose almost all its activity when temperature kept rising. When kept at 40 degrees C and 50 degrees C for 2 h, the activity of immobilized ACE remained 82% and 34% respectively, while the soluble ACE remained 52% and completely inactivated. After two kinds of enzymes were stored at 20 degrees C for one month, immobilized ACE remained 61% activity, as compared with the 20% activity residual of soluble ACE. CONCLUSION: The immobilized ACE had a better stability than soluble ACE in conditioned pH and temperature. PMID- 12903405 TI - [Construction of plasmids expressing hepatitis E virus antigen fragments and their combinations]. AB - OBJECTIVE: Several different regions of the HEV antigen were expressed in Escherichia coli and their immunological characteristics were simply evaluated. METHODS: Three coding sequences of immunodominant antigenic regions in structural protein of the hepatitis E virus (ORF2.1: 6287-6403nt, ORF2.2: 6743-7126nt, ORF3: full length of ORF3) were amplified by RT-PCR from the fecal supernatants of macaques experimentally infected with HEV. These three fragments were inserted into the pThioHisC expression vector separately or fused in line by (Gly)n short arms. After identified by DNA sequence analysis, these six recombinant plasmids were transformed into Escherichia coli, and immunology characteristics of expressed antigen fragments were evaluated by Western blotting. RESULTS: All six genes were successfully expressed in Escherichia coli. Western blotting assay showed that the recombinant proteins specifically reacted with the serum antibody from hepatitis E patient in various degrees. CONCLUSION: Artificial antigen based on strung epitopes provides a promising strategy for detection and prevention of HEV infection. PMID- 12903406 TI - [Changes of p16 gene, cyclin D1 gene, and pRb protein in human pancreatic adenocarcinoma cell lines]. AB - OBJECTIVE: To investigate the abnormalities of the relative factors of pRb-p16 cell regulatory pathway in pancreatic adenocarcinoma. METHODS: Five strains of human pancreatic adenocarcinoma cell lines were analysed by PCR and single-strand conformation polymorphism (SSCP) for homozygous deletion and mutation of the p16 gene, respectively. The gene amplification of cyclin D1 was detected by Southern blot analysis and the protein expression of pRb gene was examined by Western blot analysis. RESULTS: Homozygous deletion of exon 1 were found in two cell lines and no mutation was found within exon 1 and exon 2 in five cell lines. Cyclin D1 gene amplification was observed in one cell line and all of the five cell lines showed hyper-phosphorylated pRb protein. CONCLUSION: These findings have demonstrated that there are abnormalities of pRb-p16 regulatory pathway in pancreatic adenocarcinoma cell lines, revealing the alteration of the p16 and cyclin D1 genes. PMID- 12903408 TI - [Application of mitotic index and proliferating cell nuclear antigen index in monitoring the change of rat endometrium after administration of estrogen and progestin]. AB - OBJECTIVE: To investigate the mitotic index (MI) and proliferating cell nuclear antigen (PCNA) index in monitoring the rat endometrium during estrogen and progestin administration. METHODS: One hundred and sixty Sprague-Dolly rats were randomly divided into seven groups. The 1st and 2nd group were normal estrous (E) and ovariectomized (OVX) control. The other groups received estradiol (E2V) only or combined with medroxyprogesterone(MPA) which imitated the clinical regimen in different dosage and different time before sacrificed. Uteri sections were stained with HE and immunohistochemical procedures to detect mitosis and PCNA. RESULTS: The MI and PCNA index were similar in the luminal and glandular cells. Both markers were low in the two control groups and increased with the E2V dosage in the treatment group. But if E2V was given for 1 to 21 days, the MI increased during the first 3 days and then dropped to low level and kept to the end. PCNA index increased in the first 3 days and decreased to a low level on the 4th day and then increased gradually to a high level until the last day. In the continuous regimen, both markers decreased as the MPA dosage increased. In the cyclic regimen, The MI reached to a significant low level nearly to zero in almost each ratio. Addition of MPA for 10 days, PCNA index at the ratio of 1:4.0 could be reduced to a low level and addition for 5 days, PCNA index in each subgroup was still as high as the positive control even though the dosage of MPA increased several times to 1:8.0. CONCLUSIONS: This study suggests that PCNA index can reflect both the effect of estrogen and progestogen. PCNA index is better than mitotic index in reflecting the change of endometrium. PMID- 12903407 TI - [Expression of inducible nitric oxide synthase in human esophageal biopsies from carcinoma and precancerous lesions]. AB - OBJECTIVE: To investigate the possible role of inducible nitric oxide synthase (iNOS) in esophageal carcinogenesis. METHODS: iNOS expressions in normal epithelial cells(NC), hyperplastic cells(HC), atypical hyperplastic cells(AHC) from grade I to III, carcinoma in situ(CIS), squamous cell carcinoma(SCC), and adenocarcinoma(ADC) were detected with immunohistochemistry. RESULTS: The positive rates of immunohistochemistry staining was 0 for NC, 2.5% for HC, 4.0%, 7.5% and 2.5% for AHC grade I to III respectively, 1.4% for CIS, 8.5% for SCC and 71.4% for ADC. CONCLUSIONS: There is a high expression of iNOS in human esophageal adenocarcinomas. Frequent expression of iNOS may be a potential event in ADC carcinogenesis. There is no correlation between iNOS and SCC carcinogenesis. PMID- 12903409 TI - [Change of angiotensin system activity and the effects of corticosteroid on it in asthmatic guinea pigs]. AB - OBJECTIVE: To investigate the role of angiotensin system(ATs) and effect of corticosteroid on ATs activity at the onset of asthma. METHODS: After establishing asthmatic model of guinea pig by intraperitoneal injection with ovalbumin to sensitize and inhalation with the same sensitizer 3 weeks later to induce asthma attack, the levels of angiotensin(AT) I and AT II and the activity of angiotensin converting enzyme(ACE) were measured respectively in extractive fluid of chopped lung tissue, bronchial alveolar lavage fluid (BALF) and plasma (ACE activity in serum) in three groups of experimental guinea pigs including asthmatic group, control group and corticosteroid group. RESULTS: In the chopped lung tissue and BALF, the AT I, AT II levels and the ACE activity in the asthmatic group were significantly higher than that in the control group(P < 0.01), however, they did not differ significantly each other in plasma in three groups(P > 0.05). In corticosteroid group, the levels of AT I, AT II and ACE activity were significantly decreased in the BALF compared with that in the asthmatic group(P < 0.01). CONCLUSION: The results suggest that the lung tissue may have a local angiotensin system that may be activated and play a role in asthma attack, and the inhibition of ATs activity may be one of the mechanisms in the effects of inhaled corticosteroid on prevention and control of asthma. PMID- 12903410 TI - [Modulation of the inflammatory response through complement-neutrophil activation feedback mechanism with selenium and vitamin E]. AB - To identify the regulatory effect of sodium selenite and vitamin E on the complement-neutrophil-reactive oxygen(ROS) activation feedback (CNAF) mechanism mediated inflammatory response, we detected ROS production and complement activation in vitro tests by chemiluminescence technique and complement fixation and recognized the regulation of the inflammatory response in vivo mouse vasculitis models of skin, lung, and liver. Convincing results were observed as both in vitro and in vivo experiments showing inhibition of CNAF mechanism with sodium selenite and vitamin E could effect the reduced ROS production and complement activation. The incidence (100%) for vasculitis in control group decreased to 20%-57% in sodium selenite and vitamin E treated groups. Elucidation of the ancillary mechanism of CNAF enhancing inflammatory response is a promising area for new therapeutic developments in the modulation of inflammatory response. As in a clinical approach, a remarkable therapeutic effect with sodium selenite was observed during an epidemic episode of epidemic hemorrhagic fever in Henan province. The mortality rate of fulminant cases was reduced from 100% of untreated control cases to 36.6% by treatment with sodium selenite. The results of present studies strongly suggest that antioxidants such as selenium and vitamin E as well as others like flavonoids can exhibit a novel anti-inflammatory action via this CNAF mechanism. It is expected in the future an increasing number of patients with severe infections or inflammatory disorders in which excessive complement activation and adverse ROS production have been implicated, e. g. ischemia-reperfusion injury, severe sepsis and diverse inflammatory vascular injuries like rheumatoid arthritis, hepatitis and inflammatory bowel diseases should benefit from this newer concept guided adjuvant therapies which make use of nutrient antioxidants like selenium, vitamin E and others. PMID- 12903411 TI - [Advances in the research of the peritoneal lymphatic stomata in human]. AB - Peritoneal lymphatic stomata are small openings of the subperitoneal lymphatic vessels on the free surface of the mesothelium. The peritoneal cavity is connected with lymphatic system via these small openings which are considered to be the main passage-way that can absorb matter from the peritoneal cavity. The lymphatic stomata are claimed to be involved in many clinic procedures, such as ascites elimination; ultrafiltration failure on the continuous ambulatory peritoneal dialysis; metastasis of tumor cells from the peritoneal cavity, and so on. It was reported that the cellular factor-NO(i.e. endothelium-derived relaxing factor, EDRF) can enhance the patency of the stomata and lymphatic absorption of the stomata by stimulating guanylate way, then increasing the concentration of the cGMP, decreasing the concentration of the [Ca2+] and as a result diastole the lymphatic stomata. Some traditional Chinese medicines, which can enhance absorption of ascites, have a regulative function on the stomata by enhancing the NO concentration. PMID- 12903412 TI - [Detection of hyperdiploid in malignant cells in fine-needle aspirates from lung cancer by fluorescence in situ hybridization]. AB - OBJECTIVE: To study the numerical abnormality of chromosomes in malignant cells in fine-needle aspirates from lung cancer by fluorescence in situ hybridization (FISH). METHODS: 30 fine-needle aspirates from lung cancer (4 from lung tissues, 26 from lymph nodes) were detected by FISH with the centromere DNA probes for chromosome 7, 11, 17 and X. RESULTS: In 27 positive fine-needle aspirates the rates of hyperdiploid in chromosome 7, X, 17, and 11 were 81.5% (22/27), 77.8% (21/27), 70.4% (19/27) and 63.0% (17/27), respectively. In 3 negative fine-needle aspirates, the number of chromosomes was normal. CONCLUSIONS: (1) Interphase cytogenetic cells analysis of fine-needle aspirates by FISH is feasible and simple; (2) Hyperdiploid in malignant cells in fine-needle aspirates from lung cancer can be used as a biomarker for benign and malignant cells. PMID- 12903413 TI - [Detection of bacterial DNA in blood with three pairs of primers]. AB - OBJECTIVE: To develop a stable and sensitive method to detect gut-derived bacterial DNA present in the blood. METHODS: Blood culture and PCR were performed simultaneously when temperature of patients (n = 23) was higher than 38.5 degrees C after laparotomy. PCR was performed after DNA extraction, target genes being beta-lactosidase gene of Escherichia coli(E. coli), glutamine synthase gene of Bacteroides Fragilitas, and 16SrRNA gene of most pathogenic bacteria. DNA of standard strains of E. coli and Bacteroides Fragilitas served as positive controls, blank control as negative controls, and healthy volunteers (n = 20) served as normal controls. We performed PCR twice with part of the samples (n = 20) in order to test the repetition of this method. RESULTS: The repetition rate of PCR was 95%. Three of blood cultures were positive (13.0%, 3/23), the corresponding positive ratio of PCR was 43.5% (10/23) which was higher than that of the blood culture (P = 0.016). CONCLUSION: PCR is more sensitive than blood culture in detection of bacteremia. PMID- 12903414 TI - [Prevention and release of epidural-morphine-induced urinary retention with phenoxybenzamine and neostigmine]. AB - OBJECTIVE: To study the effects of preoperative phenoxybenzamine (o.p) on preventing epidural-morphine-induced urinary retention and effects of neostigmine on releasing it. METHODS: 80 patients who received epidural morphine to release postoperative pain were divided into 4 groups randomly and received preoperative phenoxybenzamine (group P, n = 20), neostigmine (i. m) when urinary retention happened (group N, n = 20), both drugs (group U, n = 20), and without drugs (Control, n = 20), respectively. The incidences of urinary retention and release were observed. RESULTS: No significant decrease (P > 0.05) happened after operation in the incidence of epidural-morphine-induced urinary retention both in the group P and in the group N. Neostigmine(i. m) could significantly release (P < 0.001) the epidural-morphine-induced urinary retention when phenoxybenzamine was administrated preoperatively. CONCLUSION: Preoperative phenoxybenzamine can not prevent epidural-morphine-induced urinary retention. Neostigmine can release the urinary retention only when preoperative phenoxybenzamine is used. PMID- 12903415 TI - [Chromatin structure and gene activation]. AB - Nucleosome is the basic unit of chromatin. Nucleosome repression on gene transcription is a dominant factor in the tissue or cell type specific expression on a gene from the whole genome of an eukaryote. Derepression of chromatin to an open conformation may lead to the promotion of gene transcription. The nature of chromatin remodeling, the role of histone acetyltransferase and deacetylase, the repression mechanisms of chromatin induced by the methylated CpG binding proteins are discussed in this paper. The role of chromatin in the pathogenesis of diseases has attracted broad interests from the clinicians. Cis-elements of chromatin DNA helps defining the functional segments within the genome, which provide indispensable evidences to bio-informatics, and to the linkage between genomics and proteomics. Additionally, in vitro assembly of chromatin from multiple bio-active molecules to display new functions on gene activity indicates the growing importance of integrating studies in life sciences in the coming century. PMID- 12903416 TI - [Principle of evidence-based medicine and its application in clinical trial practice for glutamine and gut permeability]. AB - Evidence-based medicine (EBM) was developed from the progress of traditional clinical model trial patterns. EBM was based on well-designed, randomized, double blind, controlled, multiple center trails in order to assess end pont parameters, prognosis parameters (as: mortality, morbidity of complications and hospital stay etc.), quality of life and cost/effect ratio. It has been accepted in developed countries and could be the guidelines for Chinese clinical trials in the future. This article summarized the principle of EBM and attached with a clinical trial sample about effects of glutamine dipeptide on gut barrier function, post operative complications and the hospital stay. PMID- 12903417 TI - [Gastric solid emptying in diabetic rats and its respond to Chinese traditional medicine]. AB - OBJECTIVE: To investigate the change of gastric solid emptying at different stages in diabetic rats, and the effect of Chinese traditional medicine(CTM) on gastric emptying. METHODS: 40 Wistar rats were randomly divided into three groups: CTM treated diabetic(15 rats), untreated diabetic(15 rats) and control(10 rats). Diabetes was induced by i.p. of streptozotocin(STZ), 65 mg/kg. CTM, 3.75 g/kg daily (consisting of Dan shen, Fu Ling mainly which functioned as Huo xue hua yu and Bu shen) was administrated in treated diabetic rats. Blood glucose and body weights were measured weekly. 12 and 24 weeks after induction of diabetes, gastric emptying was examined with scintigraphy for 240 min (30 min interval). A test meal (42 kJ) labeled with 99mTc-SC(2.96 x 10(5) Bq) was provided. RESULTS: (1) There were no significant differences in total gastric retention rate and T 1/2 among three groups at 12 weeks. (2) The lag phase of total stomach in DM-B was (41.56 +/- 8.39) min, shorter than in control's (67.50 +/- 9.09) min, (P < 0.05). (3) The proximal gastric retention rate of 12-week diabetes at 90 min were (38.2 +/- 6.2)% (vs control, P < 0.01), (16.0 +/- 7.9)% (vs control, P < 0.01), and (61.9 +/- 3.5)%; and of 24-week at 120 min were (21.3 +/- 6.8)% (vs control, NS), (48.5 +/- 8.2)% (vs control, P < 0.05, vs treated diabetes, P < 0.01), (28.3 +/- 6.4)% in treated, untreated and control group respectively. (4) There was negative correlation between blood glucose and gastric retention rate (r = -0.62, P < 0.05). CONCLUSION: (1) The proximal gastric solid emptying was impaired in diabetic rats, there was significantly accelerated in 12-week and slowed down in 24-week compared with control. (2) CTM could modify these impairment. (3) Hyperglycemia had positive correlation to gastric emptying in animal model, which might explain the clinic presentations of patients with diabetes in early stage. PMID- 12903418 TI - [Primary approach on midgastric-transverse band modifying the distribution of food between proximal and distal stomach]. AB - OBJECTIVE: To explore the physiological function of the midgastric-transverse band (MTB) in the modification of intragastric food distribution in digestive period. METHODS: 18 healthy subjects(HS) and 18 patients with functional dyspepsia(FD) participated in this study. After fasting over 8 hours all subjects took 150 ml of milk (777 J) mixed with 150 g Magnesium Sulfate, and laid supinely on 30 degrees angle with horizontal line, X-ray films were taken at 0, 30, 60, and 90 minutes after the meal, respectively. Analyzed parameters included the width of MTB, the total, proximal and distal gastric area (TGA, PGA and DGA), the ratio of proximal and distal stomach during every 30 minutes period, the correlation between MTB and PGA, MTB and DGA. RESULTS: (1) There was significant difference of MTB and DGA in every period postprandially (P < 0.05 or 0.01), by contrast, there were no significant differences of TGA or PGA between HS and FD except at TGA 90 min (P < 0.05). (2) There were close correlation between MTB and PGA at 0 (P < 0.01), 30 (P < 0.01), 60 min (P < 0.01) after the meal in HS, but only at 0 min (P < 0.01) in FD. Also close correlations between MTB and DGA at 60 (P < 0.01) and 90 min (P < 0.01) in HS, but in FD. (3) the ratios of PGA from 0 30 min and 60-90 min in FD significantly decreased respectively, however the other ratios were shown no significant difference between HS and FD during each period. (4) there were close correlations between PGA and MTB at 0, 30, 60 min after the meal in HS, however except 0 min there were no correlations in FD. By contrast, between DGA and MTB there were close correlations at 60, or 90 min in HS, no correlation in FD at any time point. CONCLUSIONS: MTB may play an important role in modifying intragastric food distribution in HS, while the dysfunction of MTB may result in the distal gastric overloading in FD, the mechanism of MTB needs further studies. PMID- 12903419 TI - [Correlation of Helicobacter pylori infection with symptoms and gastric electrical activity in functional dyspepsia]. AB - OBJECTIVE: To investigate correlation of Helicobacter pylori (Hp) infection with symptoms and gastric electrical activity in patients with functional dyspepsia (FD). METHODS: Sixty patients with FD confirmed by gastroscopy and pathology were enrolled in this study. Of them, 30 Hp positive (Hp+) and 30 Hp negative (Hp-). Ten dyspeptic symptoms were scored according to severity (0-3) and frequency (0 3), including abdominal bloating, abdominal pain, nausea, vomiting, early satiety, heartburn, and so on. Digitrapper EGG was used for recording 30 min during fasting and 60 min after a test meal (1890 kJ, C:F:P = 3.9:3.4:1). RESULTS: (1) The total symptom score (TSS) was 26.57 +/- 9.16 and 24.37 +/- 6.86 in Hp+ and Hp- groups respectively (NS). There was no significance of each symptom score between both groups, except the score of heartburn in Hp+ were higher than in Hp- group (2.30 +/- 2.04 vs 1.13 +/- 1.78, P < 0.05). (2) The percentage of normal EGG rhythm (2.4-3.7 cpm) was (64.80 +/- 4.74)% in fasting and (55.84 +/- 4.71)% in postprandial stage in Hp+ group and (64.89 +/- 5.04)%, (62.72 +/- 4.57)% in Hp- group. There was no significance of the percentage of normal EGG rhythm, power ratio(PR), and dominant frequency(DF) between HP+ and HP groups. (3) However, there was close negative correlation of TSS with normal rhythm in postprandial stage in Hp+ group (r = 0.41, P < 0.05). CONCLUSION: There may be different pathophysiologic basis in FD with or without Hp infection. Hp infection may influence postprandial gastric electrical activity. Further study needs to be investigated. PMID- 12903420 TI - [Impact of glutamine of gut permeability and clinical prognosis on the aging patients undergoing gastric-intestinal operation]. AB - OBJECTIVE: To evaluate the impact of parenteral nutrition supplemented glutamine on aging patients undergoing gastric-intestinal operation. METHODS: 30 patients above 60 years old undergoing gastric-intestinal operation, a randomized double blind protocol was designed, divided into two groups, received impact isocaloric parenteral nutrition. The study group received alanyl-glutamine [0.5 g/(kg.d)]. To observe plasma amino acids profile, nitrogen balance, intestinal permeability and clinical prognosis, examine clinical chemistry variables and observe the adverse reactions in order to find out its safety. RESULTS: The patients in both groups were comparable prior to the operation. The plasma glutamine level of study group is higher than the control group, it's cumulative nitrogen balance values were prior to the control group, L/M ratio was lower than the control group. The complications related to infection was observed more in the control group. No adverse reaction was observed in both groups. CONCLUSIONS: Ala-Gln supplemented PN improved nitrogen balance and maintained intestinal permeability, reduced complications. PMID- 12903421 TI - [An experiment of gut barrier function damage following multiple firearm injuries in a porcine model]. AB - OBJECTIVE: To study the characteristics and pathogenesis of gut barrier damage following multiple firearm injuries in porcine models. METHODS: 24 small pigs were divided into 4 groups (n = 6 each): control group(group C), group H (gunshot induced tangential fracture of parietal bone), group L (gunshot-induced comminuted fracture of bilateral femora) and group M (combined group H + L). Gastric intramucosal pH (pHi), plasma endotoxin in portal vein, and plasma D lactic acid were measured and blood samples were cultured at different time intervals after trauma. The animals were sacrificed at 72 h and ileum tissues were harvested for pathological examination and DAO measurement. RESULT: In group M at 72 h, pHi was significantly lower than that of group H and L(P < 0.01), plasma endotoxin level was significantly higher than that of group H(P < 0.01) and group L(P < 0.05), D-lactic acid level was significantly higher than that of group H(P < 0.01), and bacterial isolation rate in blood was much higher than that of group H and L(P < 0.05). Necrosis and peeling off were revealed at ileum villus top in all trauma groups, especially in group M, in which ileum DAO content also declined in most cases. CONCLUSION: Multiple traumas were prone to gastrointestinal ischemia even when hemorrhagic shock did not take place. The damage of gut barriers in multiple traumas were more severe than that in one-site trauma, which enhanced the incidence of intestinal endotoxin and bacterial translocation and trend to cause enterogenous infection. PMID- 12903422 TI - [Re-evaluation of an embolic material for aneurysm cellulose acetate polymer]. AB - OBJECTIVE: To re-evaluate the safety, effect, and pathological reactions of cellulose acetate polymer (CAP). METHODS: 30 right carotid arteries of rats were embolized and got two embolized vessels for histological study in each of the seven stages within four months. Fifteen canine aneurysmal models were created by transplantation of venous pouches. Twelve of them were occluded with CAP. Digital Subtraction Angiography (DSA) follow-ups of occluded aneurysmal models were carried out 1, 2 and 8 weeks after occlusion. One animal was sacrificed randomly at each stage for pathological study. RESULTS: Only five of the 12 aneurysmal models were completely or partially occluded within the patency of parent arteries within two months of observation after embolization with CAP. Stenosis of one branch of aneurysmal parent artery was occurred. The residual cavity of one partially occluded model artery was enlarged. Two models ruptured on four and five days after embolization. Three embolized models and their parent arteries were all occluded. PATHOLOGICAL FINDINGS: acute pathological reaction occurred within vessels embolized with CAP. Endothelial cell and basal membrane were damaged and vanished. Elastic lamina was exposed to arterial lumen and attached by thrombus layer. Fibrous cells and smooth muscle cells showed obvious degeneration. CAP and thrombus were gradually organized in two months after thrombosis. The orifices of aneurysmal models were covered with newly developed fibrous tissue and endothelial cells in two months after thrombosis. CONCLUSIONS: We think that acute chemical erosive effect of CAP was evident. A strong effect could lead to rupture of some of the CAP thrombosed aneurysmal models. Besides, stenosis or occlusion of the parent arteries could be induced in some cases after embolization with CAP. For the above shortcomings and limitations, it should be improved carefully before it is applied in clinic. PMID- 12903423 TI - [Effect of sodium azide on mitochondrial membrane potential in SH-SY5Y human neuroblastoma cells]. AB - OBJECTIVE: To study the role of mitochondrial deficiency in the pathogenesis of neurodegenerative disease by investigating the energy metabolism in a sodium azide inhibited cytochrome-c oxidase SH-SY5Y Cell model. METHODS: Human neuroblastoma SH-SY5Y Cells were exposed to sodium azide, then mitochondrial complex IV activity was assayed by microassay method; cell viability was measured by Thiazolyl blue(MTT) method; mitochondrial membrane potential (MMP) was detected by confocal microscopy and flow cytometry. RESULTS: Cultured SH-SY5Y cells were exposed to 16-64 mmol/L sodium azide for 1 hour, the mitochondrial complex IV activity decreased dose-dependently. MTT absorbance decreased does- and time-dependently in cultured nerve cells treated by 16-128 mmol/L sodium azide for 1-8 hours. After the treatment of 16 mmol/L sodium azide for 1 hour, both the fluorescence intensity of MMP and normal cell events reduced. Decrease of MMP was significant especially in cell processes. CONCLUSION: Sodium azide induced the impairment of mitochondrial energy synthesis in the cultured nerve cells which is an important cause in cell death. PMID- 12903424 TI - [Uptake of drug-containing nanoparticles in dog catotid and femoral arteries]. AB - OBJECTIVE: To study the uptake of drug-containing nanoparticles in artery for local therapy of restenosis. METHODS: Polylactic polyglycolic acid copolymer (PLGA) nanoparticles containing an antiproliferative agent 2-amino-chromone were formulated with an oil-in-water sonication emulsion/solvent evaporation technique. Arterial uptake of nanoparticles was assessed both in vitro and in vivo models of dog and rat carotid and femoral arteries. Epoxide, cyanoacrylate, fibrinogen, fibronectin, didodecyldimethylammonium bromide (DMAB), L-alpha phosphatidylethanolaine, and lipofectin were selected to modify the surface of nanoparticles to enhance arterial uptake of nanoparticles. RESULTS: The nanoparticle size ranged from 100 nm to 200 nm, the drug loading in nanoparticles was about 15%, the nanoparticle morphology was observed by scanning electron microscopy and showed spherical shape with smooth surface. Arterial uptake of nanoparticles was enhanced greatly by surface modified nanoparticle with positively charged active agent DMAB. Once nanoparticles located in artery, a high drug level could be maintained at local site for 2 days in vivo model. CONCLUSION: The primary results of animal experiments suggested that nanoparticle could be used as drug carrier of local drug delivery for treating cardiovascular diseases. PMID- 12903425 TI - [Isobutyramide activates transcription of human fetal gamma-globin gene selectively]. AB - OBJECTIVE: To examine the effect of isobutyramide synthesized in our laboratory on expression of globin genes in MEL cells transfected with the recombinant construct mu LCRA gamma psi beta delta beta MEL. METHODS: The transformant cells were cultured in the medium containing different concentrations of isobutyramide for four days. The human gamma-, beta- and mouse alpha-globin mRNA were determined by RNase protection analysis. RESULTS: The inducible and dose dependent expression of the human gamma-, beta- and mouse alpha- globin gene were observed. The induction of the human gamma-globin gene was significant stronger than that of the beta-globin gene. Under 2.5 mmol/L-5 mmol/L of isobutyramide, the induction of the human gamma-globin gene was even more effective than that of mouse alpha-globin gene. CONCLUSION: Isobutyramide can effectively and selectively activate transcription of the human gamma-globin gene at a certain degree. This function, together with the advantages of nontoxic and long halflife makes isobutyramide a promising application in the treatment of beta-thalassemia and sickle cell anemia. PMID- 12903426 TI - [Calcium activated non-selective cation channel in endothelial cell line ECV304 and inhibition of tumor necrosis factor-alpha]. AB - OBJECTIVE: To study the character of non-selective cation channel in human umbilical cord vein endothelial cell line ECV304 and the effect of tumor necrosis factor-alpha (TNF-alpha) on channel activity. METHOD: Patch clamp technique was used to record single channel and whole cell currents. RESULTS: This channel can be activated by calcium, channel conductance was (19.74 +/- 2.35) pS and its Po was 0.260 +/- 0.006 (n = 5). 100 U/ml TNF-alpha inhibited channel activity, showing that conductance reduced to (10.69 +/- 4.68) pS and Po reduced to 0.230 +/- 0.051 (n = 5). Results in whole-cell configuration were consistent with that in cell-attached patches. Currents were obviously blocked by Cl- channel inhibitor anthracene-9-carboxylic acid (A9C). In order to testify the character of this channel further, KCl and NaCl were exchanged by K(+)-Aspartate in pipette solution. Under this condition, the single channel activities as before were recorded again. Its conductance was (18.33 +/- 2.98) pS(n = 8), and TNF-alpha as well as A9C could inhibit channel activity. But this channel activity could not be blocked by another Cl- channel blocker Zn2+. In the condition of K-Asparate replaced by CsCl in solution, identical channel activity as before could not be recorded. It seemed that Cs+ could not pass through this cation channel. CONCLUSION: It is inferred that calcium-activated non-selective-action channel recorded in endothelial cell ECV304, can be inhibited by TNF-alpha and A9C. PMID- 12903428 TI - [Change of 5-fluorouracil penetration in blood-pancreatic barrier of rats after high-dose radiotherapy]. AB - OBJECTIVE: The high-dose intraoperative radiotherapy has become an important therapy to control local recur of pancreatic carcinoma. But the relevant effect of high-dose radiotherapy on chemotherapy is not clear now. METHODS: We studied the effect of high-dose radiotherapy to the penetration of 5-FU (5-Fluorouracil) in blood-pancreatic barrier of rats after the pancreatic region of the rats had treated with 10(Gy) external beam radiotherapy, the penetration ratio (PR) of 5 FU in the pancreatic tissue was investigated with HPLC assay. RESULTS: We found the PR increased with the time processing. And reached the peak on the 6th day, (PR = 0.8300 +/- 0.1662, P < 0.05). Then, the PR began to descend and was 0.7028 on the 10th day. But it was still significantly higher than control group (P < 0.05). CONCLUSION: After the treatment of high-dose radiotherapy, in addition to chemotherapy on the 4th day, and finished the course on the 10th day, the effect of radiotherapy may improve chemotherapy significantly. PMID- 12903427 TI - [Parent artery occlusion for the treatment of vertebral artery aneurysm]. AB - OBJECTIVE: To describe the endovascular treatment of posterior aneurysm with balloon occlusion of ipsilateral vertebral artery. METHODS: 10 patients aging from 12 to 52 years old with mean age 41 were performed occlusion of unilateral vertebral artery after 30 minutes test occlusion. The whole procedure was under systemic anticoagulation. The Willi's Circle was evaluated by MRA before treatment. RESULTS: No immediate complication occurred after the treatment. 10 patients were followed up for 3 months to 5 years. 8 of the 10 cases accepted MR or DSA examination. Focal infarction was found in one patient. The aneurysm was filling in contralateral vertebral artery angiography in one patient, but there was no symptom. CONCLUSION: Vertebral artery occlusion is an effective treatment to some complicated posterior circulation aneurysms, which is difficult to be clipped and coiled. PMID- 12903429 TI - [Ototoxicity of gentamicin assessed by the vestibular evoked potentials]. AB - OBJECTIVE: To assess ototoxicity of gentamicin by the vestibular evoked potentials (VsEPs) and auditory brainstem response (ABR). METHODS: 40 guinea pigs were divided randomly into 4 groups, 10 animals of group A were injected gentamicin intramuscularly 60 mg/(kg.d). 10 of group B were injected Ns intramuscularly 0.5 ml/d as control. 10 of group C were given gentamicin intratympanic 5 mg/d(0.1 ml). 10 of group D were given Ns intratympanic 0.1 ml/d as control. ABR and VsEPs were measured prior to and following the administration. RESULTS: After treatment group A 21 days, 40 days, group C 5 days, 10 days, ABR and VsEPs showed significant changes, corresponding to histopathological examination of the temporal bones. CONCLUSION: VsEPs used in assessing ototoxicity of gentamicin just as ABR. PMID- 12903430 TI - [Assay and L1 gene sequence analysis of human papillomavirus type 6 and 11 in condylomata acuminata]. AB - OBJECTIVE: To determinate the prevalence of HPV type 6, 11 in condylomata acuminata, and analyze the sequence of L1 gene and its deduced L1 protein. METHODS: PCR was used to amplify DNA sequences between E6 and E7 of HPV type 6 and 11. The PCR products were identified by the fragment length cleaved with restriction enzyme RsaI. L1 genes were amplified with high fidelity Taq, and cloned DNA sequences were detected by the dideoxynucleotide method. RESULTS: In 34 examples of condylomata acuminata, HPV11 was found in 25 examples (73.5%), HPV6 in 15(44.1%), including HPV6 + HPV11 in 6(17.6%). In comparison with the prototypes there were 7-8 point mutations in HPV11 L1 gene and 3 in HPV6L1 gene. L1 encoded protein changes showed up only in 1-3 amino acids in the former cases. CONCLUSION: Most of patients tested with condylomata acuminata are mainly infected by HPV11, the rest of them by HPV6 or by co-infection of two mixex types. Sequence analysis of L1 genes indicated that mutations were found in both HPV6 and 11, whereas deduced amino acid mutations in L1 protein were limited in HPV11 only. PMID- 12903431 TI - [Differentiation of human glioma cells BT-325 induced by sodium butyrate]. AB - OBJECTIVE: To study the differentiation of human glioma cells BT-325 induced by sodium butyrate in vitro. METHODS: BT-325 cells were treated with 1 mmol/L sodium butyrate. Flow cytometry was used to analyze the cell cycle. Cell differentiation was identified by flow cytometry and Western blot analysis. RESULTS: After 6-8 days of sodium butyrate treatment, the differentiation characters could be observed distinctively, such as the reduced cell density, the increased cell size and the marked increase in cell process formation and cell-to-cell connection. At the same time, Western blot showed that the amount of glial fibrillary acidic protein was elevated after the sodium butyrate treatment. CONCLUSION: Human glioma cells BT-325 could be induced to differentiation by sodium butyrate. PMID- 12903432 TI - [Effect of high-lipid diet on glomerular mesangial matrix in adriamycin-induced nephrotic rats]. AB - OBJECTIVE: To determined the effect of hypercholesterolemia induced by high-lipid diet on glomerulosclerosis. METHODS: Urinary protein excretion and serum cholesterol were assayed, Image analysis and techniques of pathology, immunohistochemistry, molecular biology were used to determine morphological changes in glomeruli and the production of glomerular mesangial matrixes in adriamycin-induced nephrotic syndrome (NS) and non-NS rats fed with standard chow and high-lipid chow. RESULTS: The serum total cholesterol level was significantly higher in rats with high-lipid chow in both non-NS[(2.2 +/- 0.3) g/L vs (0.9 +/- 0.1) g/L, P < 0.01] and NS[(9.5 +/- 0.2) g/L vs (2.3 +/- 0.3) g/L, P < 0.01]. The urinary protein excretion was significantly higher in the high-lipid diet rats than that in standard chow rats [(76.2 +/- 24.2) mg/24 h vs (44.8 +/- 13.6) mg/24 h, P < 0.05] in NS rats. Significant increases in the mesangial matrix and mesangial cells were observed in rats with high-lipid diet in both NS and non-NS group. Especially, more obvious pathological changes were found in NS group, such as lipid deposits and foam cell formation in mesangial areas, and progressing to focal and segmental glomerulosclerosis in some glomeruli. The immunohistochemical assay showed that the production of fibronectin was increased in the rats with high-lipid diet in non-NS group, whereas, the production of three major components (collagen IV, fibronectin, and laminin) was increased in NS group, especially in the rats with high-lipid chow. The increased expression of laminin mRNA was also detected with slot blotting in both NS and non-NS rats with high lipid chow, and it was more obvious in the rats with NS. CONCLUSION: Our findings indicated that diet-induced hyperlipidemia can lead to over-production of mesangial matrix components, and further aggravate glomerulosclerosis in adriamycin-induced nephrosis. PMID- 12903434 TI - [Surgical treatment of rectal villous adenomas: report of 49 cases]. AB - OBJECTIVE: To evaluate surgical treatment of rectal villous adenomas. METHODS: Forty-nine cases of rectal villous adenomas treated with different surgical procedures were reviewed. Twenty (40.8%) of 49 patients had Mason's operation, eleven(22.4%) had Dixon's operation, eighteen (36.8%) had transanal excision and other procedures. RESULTS: Seventeen(34.7%) of 49 were villous adenomas containing invasive carcinoma. The rates of recurrence and complication of Mason's operation were 0% and 10% (2/20), Dixon's operation were both 18% (2/11), other procedures were 33.3% (6/18) and 16.7% (3/18). CONCLUSION: Mason's operation for most cases of villous adenomas at mid and low rectum can be a safe and effective method of treatment. PMID- 12903435 TI - [Somatostatin receptor subtype 2 mRNA expression in human gastric immortalized cell line and tumor cell lines]. AB - OBJECTIVE: Inhibition of somatostatin on cell proliferation via somatostatin receptor subtype 2 (SS-R2), suggested this receptor subtype may be involved in tumor growth. Present study is to demonstrate whether SS-R2 was expressed in tumor cell lines extensively as well as in human gastric epithelia immortalized cell line. Meanwhile, the effect of p53 mutation on SS-R2 expression in gastric cancer cell line was also elucidated. METHODS: Full-length human somatostatin receptor subtype 2 cDNA was cloned by RT-PCR. Using cDNA as the probe and RT-PCR, expression of mRNA for SS-R2 in 9 cancer cell lines and human gastric immortalized cell line was detected. RESULTS: Data revealed that most cancer cell lines but a pancreatic cancer cell line expressed SS-R2 mRNA. CONCLUSION: Regardless whether p53 mutation or not in human gastric cancer cell lines, SS-R2 mRNA was transcribed. PMID- 12903433 TI - [Insulin-like growth factor-I(IGF-I), and its binding protein-3 (IGFBP-3) correlated with fetal development]. AB - OBJECTIVE: To study the relations between IGF-I, IGFBP-3 and human fetal growth. METHODS: The blood samples of maternal serum (MS) and umbilical cord serum (UCS) from 81 cases with singleton term pregnancy including 38 cases of normal pregnancy (NP), 20 cases of gestational diabetic mellitus (GDM), 23 cases of macrosomia of non-diabetic pregnancy (MNDP) were examined for IGF-I and IGFBP-3 levels by immunoassay kit from DSL, USA. RESULTS: IGF-I in MS of NP and MNDP showed a positive correlation with birth weight (BW) (r = 0.653 and r = 0.640 respectively, both P < 0.01). IGF-I of MS in MNDP was higher than that in NP (P < 0.05). IGF-I of MS and UCS in GDM were higher than that in NP (P < 0.05). IGF-I and IGFBP-3 of UCS in these three groups were significantly lower than that of MS (P < 0.001). CONCLUSION: Detection of IGF-I in MS can assess fetal intra-uterus growth, birth weight and has certain value of predicting macrosomia. IGF-I and IGFBP-3 play important roles for human fetal growth. PMID- 12903437 TI - [Alterations of the p16 gene for the carcinogenesis in pancreas]. AB - Pancreatic cancer is a common malignant neoplasm in digestive tract, which is of unknown etiology as yet. Recent advances in molecular oncology have provided explanations at the DNA level that multiple genetic changes contribute to pancreatic cancer development, in which the p16 locus of tumor tissue is nearly always altered. This tumor suppressor gene, located on chromosome 9p21, encodes for a protein that is involved in cell cycle regulation. Alterations of the p16 gene for the carcinogenesis in the pancreas involve different molecular mechanisms, including deletion, mutation or methylation. As recent anecdotal case reports that pancreatic cancer aggregates in some families, the patterns of inheritance of pancreatic cancer were studied utilizing molecular genetic techniques for some kindreds which exhibit aggregation of the cancer, and germline mutation in p16 gene have been demonstrated to predispose to pancreatic cancer. In addition, the anticancer effects of exogenous p16 gene have been shown by introduction of it into human pancreatic tumor cell lines. At the present time it is uncertain whether the p16 gene provides clinically prognostic information. Microsatellite instability was observed at one or more loci in a number of pancreatic cancers, but pancreatic cancers with mismatch repair deficiency may have an improved prognosis. PMID- 12903436 TI - [Clinical and experimental studies on condyloma acuminata]. AB - Condyloma acuminata(CA) is one of sexually transmitted diseases(STD) caused by human papillomavirus(HPV). We have done systemic study on the CA in recent years. Analyzing the morbidity of CA in our STD clinics showed that CA was in the first or second place of STD. Tissue specimens from CA were detected HPV DNA by dot blot hybridization and PCR. The result showed PCR was more sensitive than dot blot hybridization. HPV DNA were detected by PCR in tissue specimens of cervical squamous carcinoma, as well as cervical and vulvar CA. The result confirmed that HPV 16 was main type in cervical squamous carcinoma, and HPV 6 and 11 were main types in cervical and vulvar CA. To differentiate pseudocodyloma of vulva from CA, electron microscopy and HPV DNA detection were done. Certain ultrastructural changes were helpful, and detection of HPV DNA was better method in differentiating CA from pseudocondyloma. High purification of podophyllintoxin was studied and the effectiveness of podophyllintoxin was observed, compared to imported podophyllintoxin to treat CA, both cure rates were more than 80% after 3 week's therapy. Intralesional injection of different interferons for CA was effective. PMID- 12903438 TI - [Effects on the adjacent segment after spinal fusion and instrumentation]. AB - This paper reviewed the literatures about the pathology at the segments adjacent to the fusion segments. The hypertrophic degenerative arthritis of the facet joints and spinal stenosis were the most common pathologic conditions. The pathology usually occurred at the upper segment in the patients of spinal fusion; and the lower segment would also show degeneration in those of instrumentation. The reason lied mainly in the increased stress at the adjacent segment after surgery and the change of segmental motion pattern. Symptoms usually developed within the first 5 years after fusion. Diagnosis depends on the clinical manifest and X-ray examination. Surgical intervention should be indicated if the conservative therapy failed, and decompression and fusion were most commonly applied. PMID- 12903439 TI - [The identification and cloning of human ubiquitin binding enzyme cDNA]. AB - OBJECTIVE: To identify and clone the gene encoding human ubiquitin binding enzyme and study its expression spectrum. METHODS: According to the sequence of human EST, which is highly homologous to the mouse ubiquitin conjugating enzyme (E2), primers used for library screening were synthesized to screen the human fetal brain cDNA library. The gene was analyzed by making use of bioinformatics and its expression spectrum was studied by using multiple-tissue Northern blot. RESULTS: Two cDNA clones encoding human ubiquitin conjugating enzyme were isolated and identified. Both containing the ubiquitin conjugating domain, they were 88% identical in amino acid sequences and found to be isoforms of each other with only an exon excised from the short sequence. They belonged to a highly conserved, and widely expressed E2 enzyme family. Northern blot showed that they were expressed exclusively in heart, placenta, and pancreas while no transcripts could be detected in brain, lung, liver, skeletal muscle, or kidney. CONCLUSIONS: The gene encoding human ubiquitin binding enzyme is expressed under both temporal and spatial control. As a key enzyme in the degradation of proteins, ubiquitin conjugating enzymes play a central role in the expression regulation on the level of post-translation. PMID- 12903440 TI - [Alternative splicing of de novo methyltransferase gene 3b in adult and newborn mice]. AB - OBJECTIVE: To unravel the biological significance of the alternative splicing of de novo methyltransferase 3b, expression of the Dnmt3b gene in various tissues and developmental stages was investigated in postnatal mice. METHODS: RT-PCR and capillary electrophoresis were employed to analyze the alternative splicing pattern of Dnmt3b in tissues of newborn and adult mice. The results had been further reaffirmed by repeating and statistics analysis. Bioinformatics tools were used to predict the structure and hydrophobicity of the Dnmt3b exon10 coding sequence. RESULTS: Isoform with Dnmt3b exon10 was the abundant form in lung of newborn mice and in liver of both newborn and adult mice, while in other tissues of newborn and adult mice, the spliced isoform was presented as the predominant one. Peptide encoded by Dnmt3b exon10 was mainly random coil on the surface of Dnmt3b protein. CONCLUSIONS: The data demonstrate that the specific expression of Dnmt3b exists in tissues and developmental stages of postnatal mice. The alternative splicing of the exon 10 of Dnmt3b is possibly involved in the regulation of Dnmt3b's catalytic function. These results provide an insight into the developmental regulation and physiological function of the alternative splicing of the Dnmt3b gene. PMID- 12903441 TI - [Screening the trans-action factors binding to the enhancer element of rat GST-P by yeast one-hybrid system]. AB - OBJECTIVE: To investigate the diversity of the mechanisms of rat glutathione S transferase P (GST-P) gene expression regulation and the relation between rat GST P gene expression and carcinogenesis. To search the trans-action factors binding to the enhancer element of rat GST-P (glutathione S-transferase P enhancer, GPE I). METHODS: We searched the trans-action factors binding to GPE I using yeast one-hybrid system, and analyzed the DNA sequence by sequencing and computer search. RESULTS: Two positive clones pYGPE1 and pYGPE2 were obtained. The homologies of the insert sequence of pYGPE1 nucleotide and amino acid sequences with rat c-jun cDNA were 99% and 100% respectively; the homologies of the insert sequence of pYGPE2 nucleotide and amino acid sequences with rat mitochondrial adenine nucleotide translocase cDNA were 99% and 100% respectively. CONCLUSIONS: Rat c-Jun and mitochondrial adenine nucleotide translocase can bind to the GPE I core sequence, and they perhaps are the trans-action factors of GPE I. PMID- 12903442 TI - [The regulation of vitamin D3 and 9-cis-retinoic acid and their receptors on human hsp90 beta gene]. AB - OBJECTIVE: To study the effects of vitamin D3(VD3), 9-cis-retinoic acid (9-cis RA) and their receptors on the regulation of human hsp90 beta gene. METHODS: We first transfected Jurkat cells with "full length" (-1,039 bp/+1,531 bp) hsp90 beta reporter plasmid beta 1.11, then the transfected cells were treated with VD3 or/and 9-cis-RA; or beta 1.11 was cotransfected with wild type vitamin D3 receptor (VDR) or/and retinoid X receptor (RXR) expression construction and the reporter activities were assayed. Western blot was carried out to detect the protein level of hsp90 beta in the Jurkat cells that were transfected by VDR cDNA or treated with VD3 or/and 9-cis-RA. By electrophoresis mobility shift assay (EMSA), we evaluated the DNA binding activity of VDR and RXR in Jurkat cell nuclear extracts. RESULTS: With VD3 or/and 9-cis-RA treatment, the expression of hsp90 beta gene was slightly increased. The induction became prominent when both of the ligands were present. The unliganded VDR or RXR alpha all slightly repressed the constitutive expression of hsp90 beta gene, while overexpression of VDR and RXR alpha further inhibited the gene. In addition, EMSA results showed that unliganded VDR and RXR simultaneously bound to the vitamin D3 response element (VDRE) of hsp90 beta gene. CONCLUSIONS: These results suggest that VD3/VDR and 9-cis-RA/RXR co-participate in the regulation of hsp90 beta gene in Jurkat cells. PMID- 12903443 TI - [The role of MAPK pathways in hsp90 genes expression]. AB - OBJECTIVE: To study the effect of mitogen-activated protein kinase (MAPK) pathways on hsp90 genes expression in Jurkat cells. METHODS: Firstly, the phosphorylation of extracellular signal-regulated protein kinase (ERK), p38 kinase and c-Jun in Jurkat cells were detected by using Western blotting-enhanced chemically lightening (ECL) system. Secondly, Jurkat cells were transfected transiently with c-Jun N-terminal kinase (JNK) dominant negative plasmid, DN JNK1, p38 cDNA antisense plasmid, anti-p38, respectively or incubated in the present of PD98059, specific inhibitor of ERK kinase. Then the constitutive and heat-induced mRNA levels of hsp90 alpha and hsp90 beta in these cells were determined quantitatively by competitive RT-PCR. The relative mRNA level of hsp90 was compared with those in the cells transfected with mock vector (pCDNA3) or untreated cells respectively. RESULTS: In the Jurkat cells treated at 45 degrees C for 15 min, the phosphorylation of c-Jun and p38 kinase enhanced obviously, whereas the phosphorylation of ERK increased at certain degree as compared with control. Both constitutive and heat-induced expression of hsp90 genes decreased in the Jurkat cells transfected with DN-JNK1, especially for the heat-induced expression of hsp90 alpha gene which reduced to 68% of that of control. After transfected with anti-p38, the constitutive expression of hsp90 alpha gene and the heat-induced expression of hsp90 beta gene increased by 26% and 22% over the control. The constitutive expression of hsp90 alpha and hsp90 beta genes in the Jurkat cells treated with PD98059, increased to 46% and 16% of the control respectively, whereas the heat-induced expression of hsp90 genes did not change. CONCLUSIONS: Evidences showed that ERK signal pathway inhibited the constitutive expression of hsp90 genes in Jurkat cell. JNK pathway promoted both constitutive and heat-induced expression of hsp90 genes, whereas p38 pathway may not play an important role in the hsp90 genes expression. PMID- 12903445 TI - [G-protein modulates the inhibition of angiotensin II on BKCa in ECV304]. AB - OBJECTIVE: To investigate the effect of angiotensin II (A II) on maxi-conductance calcium-activated potassium channel(BKCa) activity in ECV304 cell membrane, and the possible involvement of G-protein. METHODS: Cell-attached configuration of patch clamp technique was employed to record the current of BKCa in ECV304. RESULTS: 10(-7) mol/L angiotensin II inhibited the BKCa activity. Current amplitude and open probability were decreased, open time was shortened and close time was increased. G-protein activation could eliminate the inhibition of A II. CONCLUSIONS: A II significantly decreased the activity of BKCa in ECV304, which might depolarize the membrane, and the stability of membrane might change. Thus endothelial cell dysfunction might be induced. G-protein activation was involved in the regulation of the above A II inhibition process. PMID- 12903444 TI - [Construction and expression of eukaryotic expression vector pBK-IL-1ra]. AB - OBJECTIVE: Construction of eukaryotic vector with high expression of interleukin 1 receptor antagonist (IL-1ra) for IL-1ra gene delivery. METHODS: (1) Eukaryotic expression vector pBK-IL-1ra was constructed by recombinant DNA technique, and identified by restriction mapping enzymes, PCR, and DNA sequence analysis. (2) Expressions of the IL-1ra mRNA and IL-1ra in vitro and in vivo were detected by in situ hybridization, ELISA, and immunohistochemistry. RESULTS: (1) The correct construction of pBK-IL-1ra was identified by the method above. (2) The higher levels of IL-1ra in the supernate of COS-7 cells (24 h, (P < 0.001; 48 h, P < 0.01) and the over expression of IL-1ra mRNA (P < 0.01) and IL-1ra (P < 0.05) in the synoviocytes have been detected after 48 h of the transfection with pBK-IL 1ra. (3) The over expression of IL-1ra in muscle of the mice was detected by immunohistochemistry after 4 days of pBK-IL-1ra injection (P < 0.01) CONCLUSIONS: We have successfully constructed eukaryotic expression vector pBK-IL-1ra which could highly expressed IL-1ra in vitro and in vivo, and it offers a novel possibility in the research of IL-1ra gene therapy. PMID- 12903446 TI - [Preventive and therapeutic effect of Chinese herbs naofeikang on experimental emphysema and pulmonary artery hypertension in hamsters]. AB - OBJECTIVE: To observe preventive and therapeutic effect of Chinese herbs Naofeikang on hypoxic pulmonary artery hypertension of elastase-induced emphysema hamsters and investigate its mechanism. METHODS: Hamsters for 30 days after intratracheally-instilled elastase, were kept in hypoxia environment under normal atmospheric pressure for 15 days, 50 hamsters were divided into prevention group (Prv), treatment group (T), emphysema + hypoxia (EH), and control group(N). Mean pulmonary artery pressure (MPAP) was measured before hamsters were killed. And then right ventricle hypertrophy index (RVHI) was measured, as well as circulating endothelial cells (CEC) and cells recoveries from bronchoalveolar lavage fluid (BALF) were counted. Meanwhile, pulmonary tissue changes were studied under light microscope with morphometric analysis. RESULTS: Compared with EH group, MPAP, CEC, and cells recoveries of BALF of Prv and T groups were significantly decreased (P < 0.05 or P < 0.01); Mean linear intercept and percentage of arterial media area of Prv and T groups had totally a significant difference (P < 0.01) in comparison with EH group. CONCLUSIONS: Chinese herbs Naofeikang could lower pulmonary hypertension, preserve vessel endothelial cells and lessen the inflammatory reaction in pulmonary tissue. Thereby, it could hinder the further development of emphysema and inhibit the remodeling of pulmonary small artery. PMID- 12903447 TI - [Expression of IL-2 receptors on the surfaces of L1210 cells and bioactivity assay for IL-2-toxin fusion proteins]. AB - OBJECTIVE: To confirm that L1210 cells surfaces express IL-2 receptors (IL-2R) and to establish experimental models for studying the cytotoxic effect of IL-2 toxin fusion protein in vivo and in vitro. METHODS: IL-2R on mouse leukemia L1210 cells was analyzed with immunofluorescence stain and FACS. The cytotoxic effects in vitro of these fusion proteins IL-2-PEZNM and IL-2-K-PEZNM were assayed by MTT method; mice ascitic L1210 leukemia was induced by i.p. L1210 cells and the therapeutic effect of IL-2-K-PEZNM was observed. RESULTS: 86.3% of L1210 cells expressed IL-2R on their surfaces and the relative fluorescence intensity was 51.55. Two fusion proteins IL-2-PEZNM and IL-2-K-PEZNM showed dose-related cytotoxicities to L1210 cells in vitro with the IC50 of 0.51 and 0.67 microgram/ml respectively. IL-2-K-PEZNM (5 micrograms/d) showed significantly inhibitory effect on L1210 cells and prolonged the survival time of mice by 78.6%. CONCLUSIONS: L1210 cells as a model system to measure the cytotoxic activity of IL-2-toxin fusion proteins both in vitro and in vivo is simpler, stabler, and safer model than those with other cells such as Hut102 and activated T lymphocytes. PMID- 12903448 TI - [The influences of human apolipoprotein E mutants on the learning and memory of cerebral cortex in transgenic mice]. AB - OBJECTIVE: To study the influences of the mutants of human apolipoprotein E on the function of cerebral cortex. METHODS: The Southern blot hybridization and ELISA techniques were used for identification of integration and expression of human apoE4 and apoE7 genes at the mice chromosome and in the serum of F1 transgenic mice. Then the active avoidance response test was performed to determine the alteration of learning and short term and long term memory of the transgenic mice. The serum total cholesterol (TC) and triglyceride (TG) were determined simultaneously with enzymatic procedure. RESULTS: (1) In F1 transgenic mice the human apoE genes integrated at the chromosome and expressed in their serum stably. (2) In apoE4 mice the learning and short term memorizing abilities were decreased, and in apoE7 mice the short term memorizing abilities were decreased as well. In addition, the TC and TG increased statistically. CONCLUSIONS: The overexpression of human apoE4 and apoE7 genes in transgenic mice could damage the function of cerebral cortex. It denoted that the mutations of apoE might correlate the pathogenesis of Alzheimer's disease. PMID- 12903449 TI - [The diagnostic and prognostic values of assay of serum vascular endothelial growth factor in epithelial ovarian cancer]. AB - OBJECTIVE: To detect the level of the preoperative serum vascular endothelial growth factor (VEGF) in patients with epithelial ovarian cancer, and to evaluate the correlation between preoperative serum VEGF level and prognosis of epithelial ovarian cancer. METHODS: Forty-one patients with epithelial ovarian cancer (EOC), 10 patients with borderline ovarian tumor (BOT), and 25 women with benign ovarian tumor (BET) were included as three study groups, while 20 healthy women were selected as a control group. Enzyme-linked immunosorbent assay (ELISA) was utilized to measure the serum VEGF level. RESULTS: The serum VEGF levels in patients with EOC were higher than those in patients with BOT, or with BET, or those in healthy women. The median and ranges of serum VEGF levels were as follows: EOC: median 415.0 pg/ml, range 110-2,120 pg/ml; BOT: median 312.5 pg/ml, range 100-1,250 pg/ml; BET: median 170 pg/ml, range 70-925 pg/ml; healthy women: median 165 pg/ml, range 100-735 pg/ml. When median preoperative serum VEGF levels were grouped by tumor stages, histological grade of tumor cells, histological type of the tumor, lymph node involvement, and omentum involvement, significant correlations between serum levels of VEGF and histological grade, lymph node involvement and omentum involvement were found. The patients with high serum VEGF appeared to be accompanied with poor 3-year survival. CONCLUSIONS: Preoperative serum VEGF levels seemed to be correlated with the malignant behavior of ovarian cancer, which might provide some prognostic value. PMID- 12903450 TI - [Changes of corneal endothelium in patients with primary glaucoma]. AB - OBJECTIVE: To study the density and morphology of corneal endothelium in patients with primary glaucoma. METHODS: 125 eyes of 68 patients with glaucoma were compared with 63 eyes of 32 patients without glaucoma in the same age group. Exclusion criteria included history of corneal disease, ocular inflammation, trauma, contact lens wearing and surgery. The following date were recorded: type and duration of glaucoma, glaucoma medications, and intraocular pressure (IOP) measurement. Non-contact specular microscope was performed on the central cornea, endothelial images were analyzed by computer. RESULTS: Corneal endothelium density was significantly lower in patients with glaucoma[(2386.81 +/- 289.76)/mm2] than that in controls [(2540.78 +/- 195.66)/mm2]. Cell density of primary angle-closure glaucoma(PACG) [(2262.65 +/- 338.64)/mm2] was significantly low, especially in acute PACG[(1925.16 +/- 403.38)/mm2]. The area of endothelium became bigger than normal. But the endothelium density and morphology of primary open-angle glaucoma(POAG) and non-acute PACG had no changes. No correlation was found between mean recent IOP and endothelium density. CONCLUSIONS: This study suggested that patients with acute PACG might have lower corneal endothelium density and bigger cell area than those without glaucoma in the same age group. The endothelium density and morphology of POAG and non-acute PACG were as similar as those of normal persons. PMID- 12903451 TI - [The effects of sodium nitroprusside-induced hypotension at different levels on cerebral blood flow and metabolism: a clinical study]. AB - OBJECTIVE: The clinical experiment was designed for observing the effects of deliberate hypotension at different level on cerebral blood flow and metabolism. METHODS: 30 ASA Grade I-II patients were randomly allocated to one of three groups (each group n = 10). Hypotension was induced by sodium nitroprusside in group 1[mean arterial pressure (MAP) = 55 mmHg] and group 2 (MAP = 65 mmHg). Group 3 (control) was on normotension. The changes of cerebral blood flow during hypotension were estimated according to the variability of cerebral blood flow velocity (CBFV) in the middle cerebral artery(MCA), which was obtained through transcranial Doppler ultrasonography (TCD). RESULTS: Hypotension caused a moderate decrease in CBFV, but no significant difference, in CBFV and oxygen saturation in internal jugular vein(SjO2), either among three groups or during the procedure. CONCLUSIONS: It is safe for cerebral perfusion and metabolism with deliberate hypotension at MAP = 55 mmHg. PMID- 12903452 TI - [Apoptosis of chondrocytes in cartilage]. AB - OBJECTIVE: To observe apoptosis of chondrocytes in cartilage and arthritis. METHODS: Apoptosis was detected by light and electron microscope and DNA electrophoresis, while [Ca2+]i was detected by confocal laser scanning microscope. RESULTS: Cartilage cells from osteoarthritis (OA) and rheumatoid arthritis (RA) could be seen to undergo normal differentiation after cultured for 3 days under light or electron microscope, while some chondrocytes directly isolated without culturing could give early changes of apoptosis, and normal chondrocytes in vitro after treatment with IL-1, TNF-alpha also underwent apoptosis. IL-1, TNF-alpha could induce increase of [Ca2+]i in chondrocytes. CONCLUSIONS: Our data showed that apoptosis might present in the normal cartilage as well as in OA and RA cells. IL-1, TNF-alpha could induce apoptosis of chondrocytes and [Ca2+]i increase during apoptosis. PMID- 12903453 TI - [Surgical treatment of primary intracerebral neoplasms only presenting with epilepsy]. AB - OBJECTIVE: To analyse the histopathology and the factors influencing the outcome of surgical treatment of primary intracerebral neoplasms only presenting with epilepsy. METHODS: 55 patients with primary intracerebral tumors presenting with epilepsy without other neurologic signs were retrospectively reviewed. RESULTS: 54(98.2%) cases were histologically diagnosed as gliomas among which 42(76.4%) were low-grade and 12(21.8%) high-grade. The incidence of complications for tumor removals was 32.7%. Of the 41 patients who had postoperative follow-up more than 6 months after operation, 20(48.8%) were seizure-free, 3 were rare, 4 were improved, and 14 (34.1%) had no appreciable reduction in seizure frequency. The position of the tumors was significantly correlated with the incidence of postoperative complications and the postoperative seizure control (P < 0.05). The incidence of postoperative complications for frontal and parietal tumors was higher than that for temporal tumors, while the temporal tumors had better postoperative seizure control than the frontal and the parietal ones. CONCLUSIONS: Most of the primary intracerebral tumors only presenting with epilepsy were low-grade gliomas. The position of tumors was an important factor influencing the extent of surgical removal and the postoperative seizure control. PMID- 12903455 TI - [Chemical constituents of annonaceae plants and their antitumor activities]. AB - This report is the summary of our research on the chemical constitutents and their antitumor activities of Annonaceae plants. Annonaceous acetogenins, polyoxy cyclohexenoids, and styryl-lactones from Annonaceae are the main constituents of antitumor activities. They are expected to develop a kind of new antitumor medicines. PMID- 12903454 TI - [Erythroid differentiation denucleation factor: a family of erythroid regulators for mammalian erythroid terminal differentiation/tumor suppression and the cloning of their related genes]. AB - The role of regulation of erythroid differentiation denucleation factor(EDDF) on mammalian erythroid differentiation and myeloma cell malignancy as well as cloning of their stage related genes were serially studied. Through a series of cybrid and hybridization experiments between mammalian erythroid cells and erythroleukemia or non-erythroid myeloma cells, we have demonstrated a novel family of erythroid regulators(EDDFs) in the mammalian differentiating erythroblasts which with an active peak occurred concomitantly with marked decreases in DNA, RNA and the nuclear anchoring vimentin-IF, but increased in hemoglobin synthesis in cytoplasm prior to the denucleation process during terminal differentiation. The results of cell fusion experiments verified that the supplement of regulators(EDDFs) was critical to the recovery of the originally lost features of terminal differentiation and the reversion of malignant phenotype of tumor cells. Here we showed that the erythroid regulator family EDDFs were essential regulators for the sequential expression of stage related genes of erythroid terminal differentiation, and for the redifferentiation of tumor cells to express the originally inactive globe genes, repressed the oncogenes, and vimentin-IF system, thus initiated nuclear condensation and denucleation. The EDDF gene family consisted of MEDDF, HEDDF-1, HEDRF-1, HEDRF-2 and HCNBP-1 were cloned. All were novel cDNA sequences that have been searched and registered in GenBank. They expressed varying in a stage specific manner, and acted on corresponding genes of terminal differentiation. PMID- 12903456 TI - [A novel apoptotic related gene--homocysteine-induced gene HCY-2]. AB - OBJECTIVE: To investigate the mechanisms resulting in cardiovascular diseases and birth defect. METHODS: Induced screening and differential display, Northern blot, and immunohistochemistry assay were applied to clone the homocysteine-induced gene HCY-2 and detect the expression of HCY-2 gene respectively. RESULTS: A novel HCY-induced full length cDNA--HCY-2 which encodes 142 amino acids was obtained from vascular smooth muscle cells of rats. Results of the Northern blot and immunohistochemistry assays suggest that it widely expresses in many kinds of tissues such as heart, kidney, brain, liver, and lung. In vitro, we transferred recombinant HCY-2 gene into endothelial cells and found that it could induce apoptosis and DNA injury. In vivo, HCY-2 gene could induce chicken embryo cells apoptosis and embryo malformation. CONCLUSIONS: The results indicated that HCY-2 gene might be a novel apoptosis-inducing gene and might be involved in the pathogenesis of cardiovascular diseases and birth defects induced by hyperhomocysteinemia. PMID- 12903457 TI - [Yeast one-hybrid system--one effective method studying DNA-protein interaction]. AB - Yeast one-hybrid system based on the principle of yeast two-hybrid system, is applied for isolating novel genes encoding proteins that bind to a target, cis acting regulatory element. The system may also be applied to map the DNA-binding domain of previously known, or newly identified DNA-binding proteins. Yeast one hybrid system offers maximal sensitivity because detection of the DNA-protein interaction occurs while proteins are in their native configurations. In addition, the gene encoding the DNA-binding proteins of interest is immediately available after a library screening. In this mini-review, we introduce the principle, the experimental procedures, the applications, and the merits of yeast one-hybrid system. PMID- 12903458 TI - [The research development of single nucleotide polymorphism]. AB - Single nucleotide polymorphism(SNP) is a novel set of polymorphism based on the single nucleotide variations at the genomic level, which distributes widely across the human genome and has high polymorphism information content (PIC). Now, tens of thousands SNPs have been discovered among the human genome and are introduced by many biological medicine networks. With the development of identification and analysis technique for SNP, especially the combination with DNA chips, they, as the third generation of genetic markers, are gradually fitable for fine-scale mapping of diseases, especially the complex diseases susceptibility genes and will eventually substitute for the most commonly used microsatellite markers presently. PMID- 12903459 TI - [A clinic analysis of 62 male patients with lupus nephritis]. AB - To study the difference between male and female patients with lupus nephritis(LN), we made a retrospective analysis. The clinical characteristics of 62 male patients with LN were compared with 62 female patients with LN. The result indicated the distribution of LN onset ages was different between male and female patients with LN. The incidence of LN was higher in the male than in female patients with SLE. Male patients were often misdiagnosed or diagnosed long after the onset of the disease. Arthritis and positive rate of ANA were less commonly occurred in female patients with LN. Other clinical and pathologic features of LN were similar in male and female patients. The response to treatment and prognosis of male patients with LN was relatively poor. So male patients with LN should be paid more attention to. PMID- 12903460 TI - [The diagnosis and treatment of Lyme disease: a report of six cases]. AB - We retrospectively analyzed the clinical features, diagnosis, treatment, and prognosis of 6 cases of Lyme disease from November, 1994 to December, 1998. According to our report, doxycycline, penicillin, and ceftriaxone are effective drugs for Lyme disease. The shorter the disease course is, the better the treatment effect will be. Patients in phase III usually need repeated treatment. PMID- 12903461 TI - [A beam of dawn light of study on gossypol as a safe, effective, and reversible male antifertility contraceptive--evaluation of the studies by using low dose gossypol combined with steroid hormone for male contraception]. AB - The present paper is an evaluation of the studies of two articles published in this issue of the journal which adopted a new regimen of low dose gossypol(12 mg.kg-1.d-1) combined with steroid hormones (methyltestosterone 20 mg.kg-1.d-1 and ethinyl estradiol 100 micrograms.kg-1.d-1) for 6 weeks as initial dose, and a similar low dose gossypol alone for 12 weeks as maintenance dose. Results showed that the dosage regimen could damage the epididymal sperms and onset of antifertility within 6 weeks in male rats, and prevent the incidence of the side effect of irreversible azoospermia. There was no adverse effect in viscera tissues, and the infertility could be reversible in about 6 weeks following withdrawal of gossypol. Male volunteers taking low dose gossypol (15 mg/d) could induce antifertility within 12 weeks, then followed by a maintenance dose of gossypol(10 mg/d) for 44 weeks. All of them remained infertile, and without developing hypokalemia and irreversible azoospermia. The fertility and the inducing abnormal histone-to-protamine replacement reaction as well as alteration of nuclear basic proteins could be recovered 10 weeks after withdrawal of drug treatment. These results provide a new approach for using the new regimen in clinical trial and a new prospect of gossypol as a potential male contraceptive. PMID- 12903462 TI - [Combined use of steroid hormone and low dose gossypol for antifertility and its mechanism in rats]. AB - OBJECTIVE: To study the feasibility and the morphological mechanism of combined use of steroid hormone (methyltestosterone and ethinyl estradiol) and low dose gossypol for antifertility in adult Wistar rats. METHODS: We used methyltestosterone 20 mg/(kg.d), ethinyl estradiol 100 micrograms/(kg.d), and gossypol 12 mg/(kg.d) to feed male rats for 6 weeks via gastric intubation to induce infertility, then used gossypol 12 mg/(kg.d) alone for another 12 weeks to maintain infertility. Testicular sperm counts, cauda epididymal sperm motility, and sperm silver staining were used to determine infertility and morphology of sperm at the end of the two phases. Testicular morphology of the rats was observed from HE and PAS staining of paraffin sections. F1 generation of the rats would be examined in a series of behavioral tests after their 6-week recovery periods. RESULTS: There was no substantial change in testicular sperm counts of drugs-treated rats both for 6 weeks and 6 plus 12 weeks compared with that of control. But cauda epididymal sperm motility of drugs-treated rats went down to zero at the end of two phrases. The result of sperm silver staining showed that epididymal sperm were all deleterious of either segregation of sperm nucha from sperm head, or separation in annular granule of sperm tail. The observation of testicular morphology suggested that spermatogenesis is only partially affected. The ratios of stages II-III (9.3 +/- 3.4)% and stages IX-X (3.6 +/- 0.6)% were remarkably dropped, but that of stages IV-VI (36.7 +/- 5.0)% obviously ascended comparing with (26.8 +/- 1.7)%, (6.0 +/- 0.4)%, (24.5 +/- 2.7)% of control at the end of 6 plus 12 weeks. Moreover, no adverse effect was found in viscera tissue sections. CONCLUSIONS: Steroid hormone made the procedure of spermatogenesis slower and low dose gossypol caused all sperm lose their activity in epididymis. Both affect the process of spermatogenesis from different endpoints and successfully induce infertility in short term. Low dose gossypol not only executes its antifertility function in epididymis, but also affects the quality of spermatozoal production in testis by impacting the procedures of both acrosomal formation and spermatozoal elongation, so it can completely maintain infertility in long term. The infertility was reversible that the infertile rats can be recovery in about 6 weeks. Their F1 generation were proved to be normal through a set of behavioral tests. PMID- 12903463 TI - [Effect of low dose gossypol treatment on male sperm nuclear basic protein]. AB - OBJECTIVE: Study on human sperm nuclear basic proteins after low dose of gossypol treatment. METHODS: Twenty-three men with normal fertility were divided into two groups. The experimental group included 15 men which were treated daily with gossypol (15 mg/d for 12 weeks and treated with 10 mg/d for 32 weeks continuously), 8 men were used in the control group. Semen was collected at 1, 4, 8, 12, 16, 24, 36 and 44 weeks after gossypol treatment and 10 weeks after stopping treatment. Total nuclear basic protein (TNBP) was extracted and TH/TP, HP1/(HP2+HP3) ratios were determined by scanning microdensitometry following electrophoresis of TNBP in polyacrylamide gels. RESULTS: The TH/TP ratio of control and the gossypol treated group were 0.17 +/- 0.07 and 0.63 +/- 0.79(P < 0.05); The HP1/(HP2+HP3) ratio of control and gossypol treated group were 1.05 +/ 0.21 and 1.47 +/- 0.18 (P < 0.01). These two parameters were returned to normal after 10 weeks of stopping gossypol treatment. CONCLUSIONS: The interruption of histone-to-protamine replacement reaction (HPRR) and alteration of nuclear basic proteins induced by low dose gossypol treatment might lead to infertility. Changes of HPRR and nuclear basic protein were reversible after stopping gossypol treatment. PMID- 12903464 TI - [Male contraception of triptonide and its function mechanism]. AB - OBJECTIVE: The male contraceptive mechanism in rats of triptonide, isolated from Multiglycosides of Tripterygium wilfordii (GTW), was investigated. METHODS: The dosage of triptonide was 200 micrograms/(kg.d), fed for 8 weeks via gastric intubation. Sperm counts, sperm motility, and sperm silver staining were used to test the antispermatogenic effect of triptonide; the study of morphology was learned from paraffin sections of testis and visceras of triptonide-treated rats; protamine was abstracted from testis by electrophoresis; using antisense mRNA of cyclin D1 and Cdk4 as prodes, we examined the expression of cyclin D1 and Cdk4 through hybridization in situ. RESULTS: Sperm counts (26.43 +/- 3.90) x 10(6)/g and sperm motility(66.0 +/- 6.8)% of triptonide-treated rats were substantially different from those of control, (41.15 +/- 5.51) x 10(6)/g (P < 0.01) and (88.0 +/- 2.0)% (P < 0.001). Paraffin sections of testis of triptonide-treated rats stained by PAS showed that the number of spermatids per 100 sertoli cells (9.15 +/- 0.33) was remarkably decreased compared with control's (10.73 +/- 0.38) (P < 0.001). At the same time, delaying sperm-released process was observed, which affects the nature course of sperm. And the extended vacuoles of vary sizes, the segregation of cells in lumens were also found in the cross-section of testis of rats treated with triptonide. In addition, the contents of protamine (4.6 +/- 2.2)% was extremely decreased, compared with the control's (40.6 +/- 7.2)% (P < 0.001). By using antisense mRNA of cyclin D1 and Cdk4 as prodes, we detected that the expression of the genes was notably increased in spermatids. CONCLUSIONS: The results suggest the antispermatogenic effect of triptonide is very well and it mainly plays its role on spermatids by increasing the expression of cyclin D1 and Cdk4, meanwhile, decreasing the biosynthesis of protamine. The appropriate target region of triptonide makes itself most potency to be a suitable male antifertility drug. PMID- 12903466 TI - [G-protein potentiates the activation of TNF-alpha on calcium-activated potassium channel in ECV304]. AB - OBJECTIVE: Observe the effect of tumor necrosis factor-alpha (TNF-alpha) on calcium-activated potassium channel in ECV304 and the possible involvement of G protein mediation in the action of TNF-alpha. METHOD: Using the cell-attached configuration of patch clamp technique. RESULTS: (1) the activity of high conductance calcium-activated potassium channel (BKca) was recorded. Its conductance is (202.54 +/- 16.62) pS; (2) the activity of BKca was potentiated by 200 U/ml TNF-alpha; (3) G-protein would intensify this TNF-alpha activation. CONCLUSIONS: TNF-alpha acted on vascular endothelial cell ECV304 could rapidly activate the activity of BKca. Opening of BKca resulted in membrane hyper polarization which could increase electro-chemical gradient for the resting Ca2+ influx and open leakage calcium channel, thus resting cytoplasmic free Ca2+ concentration could be elevated. G-protein may exert an important regulation in this process. PMID- 12903465 TI - [Coexpression of the preS1(1-42) and the Core(1-144) antigen of HBV in E. coli]. AB - OBJECTIVE: To study the therapeutic T cell vaccine for the treatment of chronic hepatitis B. METHODS: The genes of HBcAg (1-144) and preS1 Ag (1-42) were amplified and fused by PCR. This fusion gene was inserted in the prokaryotic expression vector pET-11d and expressed in E. coli. RESULTS: It was showed by SDS PAGE that the protein molecular weight of the coexpression product was about 20,000, twenty percent of all bacteria protein. The monoclone antibody against Core and preS1 antigen could react with this fused protein by Western-blot technique respectively. The fused gene was verified by sequencing. Under the immune electron microscopy, this fused protein is a typical particle of HBcAg but in an aggregated form. CONCLUSIONS: The results may aid for studying T cell immunotherapeutic vaccine to chronic hepatitis B. PMID- 12903467 TI - [Role of nitric oxide in excitotoxicity and memory impairment induced by kainic acid]. AB - OBJECTIVE: The role of nitric oxide (NO) in excitotoxicity of kainic acid (KA) in the brain was investigated. METHODS: Adult female rats were bilaterally intra caudate nucleus injected with KA. The nitric oxide synthase (NOS) was determined by measuring NO2- concentration produced from homogenized brain tissue in the enzyme response system at three time point after surgery. Another groups of rats were daily administrated (i.p.) L-NAME(10 mg/kg) or 7-NI(25 mg/kg) at 2nd to 6th day after intra-caudate KA injection. Behavioral tests for learning and memory were started at 10th day after surgery. RESULTS: NOS activity in caudate nucleus remained unchanged in 6-8 hours after surgery. It increased significantly in 3 days and slightly increased in 5 days after surgery. Both L-NAME and 7-NI ameliorated memory impairment induced by KA injection in passive avoidance response task. Only L-NAME improved KA-induced deficit in active avoidance response behavior. CONCLUSIONS: This study gives further evidence that NOS activation and excessive NO may mediate KA-induced excitotoxicity. Early and repetitive use of NOS inhibitors shows some protective effect on memory deficit induced by KA excitotoxicity. PMID- 12903468 TI - [Gastro-pyloro-duodenal motor cooperation in patients with dyspepsia]. AB - OBJECTIVE: To investigate the role of the midgastric transverse band(MTB) and gastro-pyloro-duodenal motility in functional dyspepsia (FD) and diabetic dyspepsia (DD). METHODS: Healthy subjects, patients with FD and DD participated in the study. Of them, 65 subjects received radionuclide gastric emptying, 48 complex ultrasonograph, and 35 gastro-pyloro-duodenal manometry. RESULTS: (1) MTB was (7.01 +/- 1.31), (3.39 +/- 0.21) cm in FD and DD, which was significantly wider than that in HS group [(2.42 +/- 0.16) cm, P < 0.01, P < 0.001]. (2) The half gastric emptying time (t 1/2) in FD and DD were prolonged, gastric remained rate at 120 min increased, the gastric emptying velocity reduced when compared with HS. (3) The gastric occlusive and non-occlusive peristaltic contractions could be clearly shown under ultrasonograph. The percentage of emptying circle > 10 seconds was reduced in FD and DD. (4) There was decreased number of migrating motor complex phase III in FD and DD, when compared with HS. CONCLUSIONS: MTB is an area with physiological function, which affects intra-gastric food distribution. Patients with FD and DD showed delayed gastric emptying associated with decreased gastric motor activity and antro-pyloro-duodenal in-cooperative function. There may be different pathogenesis in FD and DD. Further study needs to be performed. PMID- 12903469 TI - [Electrogastograph during interdigestive and digestive stages in patients with functional dyspepsia]. AB - OBJECTIVE: To investigate the relationship between dynamic change of electrogastrogram (EGG) with motility index of antrum during migrating motor complex (MMC) cycle in healthy subjects and functional dyspepsia, and to verify whether a 100% coupling of electrical and mechanical events existed after meal. METHODS: Eleven healthy subjects (HS) and 14 patients with functional dyspepsia (FD) were studied. An 8-channel Dent-sleeve catheter was positioned under fluoroscopic guidance across antropyloroduodenal region and bipolar electrodes were attached to the skin corresponding to the location of antrum. Antropyloroduodenal pressure waves and electrical signals were transferred simultaneously to PC Polygraph (CTD) and recorded for 3 hours preprandially and 2 hours after ingestion of 40 g solid meal (336 kJ). The power of EGG and motility index (MI) of antral contractions were calculated as the area under curve (AUC) by a build-in computer program. RESULTS: The power increases of EGG increased significantly in phase II (P < 0.01), phase III (P < 0.01) and after meal (P < 0.01) compared with phase I in HS, however, this dynamic increase was not found in FD. The power of EGG was positively correlated with the motility index of antrum both in HS (r = 0.87, P < 0.01) and in FD (r = 0.79, P < 0.01) during the interdigestive period. A significant increase of EGG power after meal was found in HS, which corresponded to the degree of phase III and kept stable for the whole postprandial period. In contrast, power increase was not found in FD except during 30 min after test meal. CONCLUSIONS: Prolonged cutaneous EGG recording can reflect the periodic variation of MMC cycle. EGG response in HS but not in FD after meal is characterized as persistent increase which is corresponded to phase III. PMID- 12903470 TI - [Changes of handling of calcium in mesenteric vascular smooth muscle cells of calcium overload rat]. AB - OBJECTIVE: To investigate if there existed abnormal handling of calcium in vascular smooth muscle cells (VSMC) in calcium overload rat (CaOMV). METHODS: Cytosolic and nuclear calcium ion handling in mesenteric VSMC both in calcium overload rat and in Wistar rat were studied by using laser scanning confocal microscopy together with calcium-sensitive-dye Fluo-3/AM. RESULTS: The results showed that both nuclear and cytosolic calcium level ([Ca2+]n, [Ca2+]i) in mesenteric VSMC in calcium overload rat under resting condition was not higher than that in normal rat. However, [Ca2+]n and [Ca2+]i in CaOMV increased more significantly than that in normal rat when induced by drugs such as KCl, Bay k8644, Ang-II, IP3, CHCl3, and CPA. CONCLUSIONS: (1) There exists abnormal handling of calcium in CaOMV because of it's overreaction to stimulators, which indicate that intracellular and extracellular calcium overload coexist in age related arteriosclerosis; (2) Not only Ca2+ gating system of plasma membrane but also calcium pool in CaOMV are irregular; (3) Cytosolic calcium as well as nuclear calcium are handled improperly in CaOMV. PMID- 12903471 TI - [Mechanisms of opioid receptor-induced elevation in intracellular calcium by confocal laser scanning microscopy]. AB - OBJECTIVE: To determine the acute and chronic effects of opioid receptor agonists on the intracellular free calcium concentration ([Ca2+]i) in NG-LNCXiNOS cells, stably expressing iNOS gene, and regulation of G-protein on opioid-induced response in [Ca2+]i. METHODS: A single cell [Ca2+]i is measured by confocal laser scanning microscopy using Ca(2+)-sensitive dye Fluo-3 as an new calcium fluorescent probe. RESULTS: DPDPE(D-Pen2, D-Pen5-enkephalin), a delta-opioid receptor agonist, and morphine acutely induced the increase in [Ca2+]i of NG LNCXiNOS cells. The elevation in [Ca2+]i by DPDPE could be abolished with naloxone. Pretreatment of the cells with pertussis toxin (PTX) at 100 ng/ml for 24 hours almost completely blocked morphine-evoked response. In contrast to acute effect of opioid agonists on [Ca2+]i, the cells exposed to 1 mumol/L DPDPE or 10 mumol/L morphine for 48 hours also appeared to raise [Ca2+]i. However, the elevation in [Ca2+]i was not greater than that caused by acute effect of DPDPE or morphine. After cell "withdrawal" was precipitated by the addition of 10 mumol/L naloxone, the increase in [Ca2+]i could further be intensified. CONCLUSIONS: The opioid agonist-induced increase in [Ca2+]i is mediated by opioid receptor and regulated though PTX-sensitive G-protein. The attenuation of this response in chronically treated cells with opioid agonist is associated with receptor desensitization. PMID- 12903472 TI - [DNA damage and c-myc gene expression of NIH3T3 cell induced by superoxide anion]. AB - OBJECTIVE: To study the DNA damage, membrane lipid peroxidation, and c-myc gene expression of NIH3T3 cell induced by superoxide anion(O2.-). METHODS: The superoxide anion(O2.-) was produced by reactive system of xanthine-xanthine oxidase (X-XO) and the DNA-EB complex, malondialdehyde (MDA) content and expression of c-myc gene were measured by means of fluorescence, thiobarbituric acid (TBA) assay and DNA-RNA situ hybridization with digoxigenin labeled probes respectively. RESULTS: The O2.- of high concentration(X: 20 micrograms, XO: 2 x 10(-3) U) reacted with isolated DNA from NIH3T3 cell directly which caused DNA damage obviously. When the O2.- of high concentration reacted with whole NIH3T3 cell, the amount of MDA increased and fluorescence intensity of DNA-EB complex decreased which mean lipid peroxidation of membrane and gene damage of the cell respectively. When the producing O2.- of high concentration reactive system existing FeSO4, the phenomena mentioned above showed obviously. Enough amount of catalase showed the inhibit effects, but it could not completely inhibit the DNA damage of whole cell. The c-myc gene expression was observed in the cell treated with O2.- of middle (X: 4 micrograms, XO: 4 x 10(-4) U) and high concentration, but it was not observed in the cell treated with O2.- of low concentration(X: 0.4 micrograms, XO: 4 x 10(-5) U). CONCLUSIONS: DNA damage of the whole cell induced by extracellular O2.- was able to act through the pathway of membrane lipid peroxidation in which hydrogen peroxide is important and the other pathways such as signal transduction of the cell might also be existed. The expression of c-myc gene of NIH3T3 cell induced by O2.- was related closely to the concentration of O2.-. PMID- 12903473 TI - [Transgenic mice model of human hepatitis B virus x gene]. AB - OBJECTIVE: To set up a human hepatitis B virus x (HBx) gene transgenic model. METHODS: The model was set up by microinjection, the integration and expression of the founders were identified by molecular hybridization. RESULTS: A 1,800 bp DNA fragment containing the human HBx promoter and genomic DNA was microinjected into the male pronucleus of the fertilized eggs of mice. Three hundred and thirty eight eggs survived from microinjection were transplanted into the oviduct of 16 pseudopregnant female, 12 pseudopregnant mice got pregnant and gave birth to 74 pups. Genomic DNA were extracted from the tails of the pups, Southern blot hybridization was used for determination of the integration of human HBx gene. The 1,800 bp microinjected fragment was used as the probe, 17 mice (founders) were identified of the integration of human HBx gene. Total RNA were extracted from the livers of the founders, all 17 founders were identified of the expression of human HBx gene by using Northern blot hybridization. CONCLUSIONS: These results may provide animal models for studying on the correlation between chronic HBV infection and the development of hepatocellular carcinoma. PMID- 12903474 TI - [Clinical analysis of glucose dysbolism in 162 cases with Cushing syndrome]. AB - OBJECTIVE: To understand the characteristic of steroid diabetes by the analysis for clinical data of 162 cases with Cushing syndrome (CS) proved by pathological report. METHODS: Retrospective analysis for the data of CS about age of CS onset, duration of CS, body mass index (BMI), 24 hours urine free cortisol (24 h UFC), pathological report, fasting blood glucose (FBG) pre- and post-operation, family history of diabetes, treatment of glucose dysbolism (GD) pre- and post-operation. RESULTS: The onset age and duration of CS in GD group was obviously earlier and longer than normal glucose (NG) group. BMI and 24 h UFC were no difference between two groups. Besides adrenal adenocarcinoma, no difference was found in types of pathology. FBG of diabetes pre-operation was obviously lower than post operation. After operation, most diabetes need not on drug treatment. Only one secondary diabetes has family history. CONCLUSIONS: Steroid diabetes easily occurs in patients who is senility and suffer longer course of CS. No relation to diabetic family history, concentric obesity, the level of UFC and benign types of pathology. Therapeutical emphasis of steroid diabetes is radical cure of CS. PMID- 12903475 TI - [Effect of weight loss on adipose tissue distribution and insulin sensitivity]. AB - OBJECTIVE: To discuss the effect of weight reduction on fat distribution and parameters of insulin sensitivity. METHODS: 12 Caucasian women with simple obesity, aged 21 to 65 years, were treated by low caloric and high protein diet for four weeks. A series of examinations were taken before and after the treat, using computerized tomography for visceral and subcutaneous fat, using euglycemic insulin clamp for parameters of insulin sensitivity. RESULTS: Weight loss was (6 +/- 2) kg (2-11 kg). body mass index (BMI), waist, total fat, visceral fat, and subcutaneous fat reduced significantly. However waist-hip rate, visceral-total fat rate, and subcutaneous-total fat rate did not change significantly. Fasting serum insulin, and fasting insulin-glucose rate decreased significantly. Insulin metabolic clearance rate and, insulin sensitive index increased significantly. Fasting peptide C did not change significantly. CONCLUSIONS: The low caloric and high protein diet can reduce visceral and subcutaneous fat. Weight loss can improve insulin sensitivity and increase insulin metabolic clearance rate. PMID- 12903477 TI - [Follow-up study of total condylar knee arthroplasty with three types of Chinese knee prosthetics]. AB - OBJECTIVE: To clinical evaluate the results of total condylar knee arthroplasty using three types of Chinese knee prosthesis. METHODS: Hundred and two knees (in 92 patients) arthroplasty were performed in our department during the 14 years (April 1981-November 1995) were analyzed. The follow-up period averaged 64.4 months. Eight cases had died and 14 cases were lost of follow-up, leaving 80 knees in 70 patients for clinical evaluation. The clinical results were evaluated according to Hospital for Special Surgery Rating System. RESULTS: The average preoperative score was 47.65 and the postoperative score was 90.23. Almost all patients achieved significant pain relief after operation. Sixty-one of the 70 patients (87%) reported that they satisfied or very satisfied with the results of surgery. There were 6 failures. One was due to deep infection, one with loose patellar component, and four directly related to either improper selection or technical reasons. CONCLUSIONS: Total condylar knee arthroplasty with Chinese knee prosthetics gives a satisfactory grade of pain relief and movement of the operated knee. Chinese knee prosthetics is safety and durable. PMID- 12903476 TI - [A clinical study on coincidence with hyperthyroidism and thyroid carcinoma]. AB - OBJECTIVE: A retrospective study has been carried out to evaluate the prevalence of coincidence with hyperthyroidism and thyroid carcinoma. METHODS: 394 patients underwent surgery for hyperthyroidism and 245 patients suffered from thyroid cancer were chosen for the study in our hospital from January 1983 to June 1998. RESULTS: Thyroid cancer and hyperthyroidism coincided in 12 patients. The incidence of thyroid cancer was 3.0% (12/394) in hyperthyroidism, and the incidence of hyperthyroidism was 4.9% in thyroid cancer. There were 7 female and 5 male, with a diffuse goiter (n = 3), a diffuse goiter with a cold nodule (n = 3), multinodular goiter (n = 6). Among the 12 patients, 7 patients had an occult thyroid cancer with a diameter of 1 cm or less, most of them with papillary carcinoma, less frequently had metastases, 4 patients had thyroid cancer with a diameter of 3 cm or more, and 3 patients had metastases. CONCLUSIONS: The prevalence of coincidence with thyroid cancer and hyperthyroidism is more than that of thyroid cancer in population. Diagnostics for exclusion of thyroid cancer is required carefully even in the presence of hyperthyroidism. PMID- 12903478 TI - [Effect of macrophage nitric oxide on the ultrafiltration failure of long-term peritoneal dialysis]. AB - OBJECTIVE: The regulation of macrophage nitric oxide on peritoneal lymphatic stomata was studied for revealing the mechanism of ultrafiltration failure during long-term peritoneal dialysis. METHODS: (1) The model of peritoneal dialysis was created by using peritoneal dialysate; (2) Dynamic measurements of nitric oxide (NO) were made during peritoneal dialysis and its cessation; (3) The pathological change of the peritoneal mesothelium in different dialysis time was observed by scanning electron microscope (SEM); (4) Dynamic changes of the peritoneal lymphatic stomata were studied during this experiment by using a computer image processing system attached to SEM. RESULTS: Numerous macrophages went into the peritoneal cavity through the lymphatic stomata to form a lot of milky spots during peritoneal dialysis. A great quantity of NO produced by macrophages damaging mesothelial cells and increasing numbers and density of lymphatic stomata. After 40 days of peritoneal dialysis, the diameters of the lymphatic stomata were significantly increased in Bieffe group (P < 0.05). During cessation of peritoneal dialysis, few macrophage milky spots were observed and NO quantity was gradually decreased. Then the mesothelium damage began to repair and the peritoneal lymphatic stomata tended to become normal. CONCLUSIONS: Our data demonstrate that a great quantity of NO produced by macrophages would damage peritoneal mesothelium and relax lymphatic stomata which enhance lymphatic reabsorption from the peritoneal cavity so as to make ultrafiltration failure on long-term peritoneal dialysis. PMID- 12903480 TI - [Stage-dependent expression of androgen binding protein mRNA in sertoli cell of rat testis]. AB - OBJECTIVE: To investigate the variation of rat androgen binding protein (ABP) mRNA in sertoli cell of rat testis during spermatogenesis. METHODS: In situ hybridization was performed with digoxin-labeled rat ABP cDNA probe on frozen testis tissue sections from SD rats. The expression of ABP mRNA was quantified by using laser density scanning system. RESULTS: It was shown that ABP mRNA levels were low from stage I to stage VI, increased to the peak level at the stages VII VIII, and then declined rapidly at stages IX-XI. Thereafter, ABP mRNA levels increased again at stages XII-XIV, and then decreased to the lowest level at stages I-VI. CONCLUSIONS: ABP mRNA level was stage-dependent during the spermatogenic cycle. PMID- 12903479 TI - [Effect of propofol and etomidate for anesthesia induction on plasma total cortisol concentration]. AB - OBJECTIVE: To investigate the effect of propofol and etomidate for anesthesia induction on adrenocortical function. METHODS: The plasma total cortisol concentration was determined in 20 patients who were treated with propofol or etomidate. RESULTS: After propofol induction, the plasma total cortisol level decreased from (308.7 +/- 27.4) nmol/L to (267.7 +/- 31.2) nmol/L at post medication 2 hours, increased to (400.2 +/- 26.9) nmol/L at postmedication 5 hours and restored to premedication level at postmedication 24 hours [(306.4 +/- 35.4) nmol/L]. After etomidate induction, the plasma total cortisol level decreased from (309.1 +/- 36.6) nmol/L to (115.9 +/- 29.7) nmol/L at postmedication 2 hours, maintained (171.1 +/- 34.7) nmol/L at postmedication 5 hours and restored to premedication level at postmedication 24 hours [(311.8 +/- 46.2) nmol/L]. CONCLUSIONS: Propofol induction has little effect on adrenocortical function whereas etomidate induction inhibits adrenocortical function. PMID- 12903481 TI - [Effect of 5,7-dihydroxytryptamine on the distribution pattern of calcitonin gene related peptide in different motoneuron pools]. AB - OBJECTIVE: The effect of 5,7-dihydroxytryptamine (5,7-DHT) on distribution pattern of calcitonin gene-related peptide (CGRP) in different motoneuron pools was studied in rats. METHODS: Cholera toxin B subunit coupled with colloidal gold (CB-Au) retrograde labeling combined with CGRP immunocytochemistry technique was mainly used. RESULTS: In control group, there was a different distribution pattern of CGRP-LI content in soleus motoneuron (SOL-Mn) and extensor digitorum longus motoneuron (EDL-Mn) pools. SOL-Mn pools had a higher ratio of neurons lacking CGRP-LI/weak CGRP-LI and a lower proportion of strongly CGRP-LI labeled ones. Comparing with control, there was a marked shift to increasing of CGRP-LI content in SOL-Mn pool and slight decrease in EDL-Mn pools 14 days after 5,7-DHT. All the SOL-Mns were CGRP-LI positive and the ratio of CGRP-LI strongly positive Mns significantly increased (P < 0.001) while 5,7-DHT did not increase the ratio of CGRP-LI strongly positive Mn in EDL-Mn pool. CONCLUSIONS: Except muscular motor activity, the different distribution pattern of CGRP-LI content in SOL-Mn and EDL-Mn pools is related to different 5-HT inputs. SOL-Mns may receive more 5 HT inputs. PMID- 12903482 TI - [Effect of antisense oligonucleotides on the TXA2 production of macrophages stimulated by endotoxin]. AB - OBJECTIVE: To select efficient fragments which can inhibit gene expression of thromboxane synthase and production of thromboxane A2. METHODS: Rat peritoneal macrophages were incubated with oligonucleotides (ODNs) and endotoxin (which was composed of lipopolysaccharide) for 24 hours. The TXB2 content was determined by RIA. Thromboxane synthase (TXS) mRNA expression was measured with RNA dot hybridization. RESULTS: The concentration of TXB2 in ODN I group was (69.7 +/- 10.9) pg/microgram cell protein, lower than those in control group (100.6 +/- 28.6) pg/microgram cell protein (P < 0.05, n = 4), TXS mRNA expression level in ODN I group was decreased as compared to those in control group. Neither in ODN II nor in ODN III group were there any changes on TXB2 contents and TXS mRNA levels. CONCLUSIONS: ODN I can inhibit the production of TXA2 and the gene expression of thromboxane synthase from macrophages stimulated by endotoxin. PMID- 12903483 TI - [Osteopontin of skin and hair follicle of rat]. AB - OBJECTIVE: To investigate the changes of osteopontin mRNA of skin and hair follicles of embryonic and postnatal rats. METHODS: The follicular cycles of rat hair were determined firstly and then the distribution and quantity of osteopontin mRNA on the paraffin sections of skin and hair follicles of Wistar rats were determined, aged from embryonic 14-18 days and postnatal 1-61 days, were investigated by radiolabel in situ hybridization technique. RESULTS: The results indicated that when the rats were 18 days old, the signals of osteopontin mRNA in dermal papilla (DP) of hair follicles were observed. The signals of osteopontin mRNA in DP showed a regular changes in the hair cycles: the signal of osteopontin mRNA appeared in the catagen phase and the peak level was in the middle stage of catagen, but it totally disappeared when the follicles entered into telegen and anagen phases of hair cycles. Osteopontin message was mainly accumulated in follicular dermal papillary cells, and in corresponding connective tissue sheath and neighboring dermal macrophages, but no signal of osteopontin mRNA was detected in any other skin and hair follicular cells else where. CONCLUSIONS: These results suggest a possible role of osteopontin in up regulating growth and differentiation of dermal papilla during the catagen phase of the hair follicles. PMID- 12903484 TI - [Detection of proteolytic activity of hepatitis C virus NS3-4A protease using enzymelinked immunosorbent assay]. AB - OBJECTIVE: To establish an ELISA method for detecting proteolytic activity of HCV serine protease for screening inhibitors against HCV. METHODS: HCV recombinant plasmid pMAL-c2/NS3-4A was transformed into the E. coli strain K12 TB1. Maltose binding-protein (MBP)-NS3/NS3-4A fusion protein expression were induced by adding isopropyl-beta-D-thiogalacto-pyranoside (IPTG) and purified by affinity chromatography. The immunological activities of the fusion protein were analyzed by Western blot. A peptide substrate was used to analyze the biological activity of the fusion protein. The hydrolyzed product was treated with sodium iodacetate and labeled with digoxigenin, then adding immunological anti-digoxigenin-alkaline phosphatase conjugate to detect the protease activity by colorimeteric reaction. RESULTS: The purified MBP-NS3 and MBP-NS3-4A proteases were identified as 112,000 u and 116,000 u protein by Western blot. HCV NS3/NS3-4A protease showed substrate cleavage activities by ELISA. Coefficients of variation (CV) of ELISA in a lot and among the lots were 4.16% and 7.52% respectively, the P/N was 3.63 under the best experimental conditions determined by L9 (3(4)) factorial design. The method confirmed that 8.36 mumol/L 1,4-naphthoquinone had 50% inhibitive activity on HCV serine protease. CONCLUSIONS: We have established a simple and rapid ELISA method with stable repeatability for detecting proteolytic activities of HCV NS3-4A protease, which might be used for screening and studying of specific inhibitors of HCV serine protease NS3-4A. PMID- 12903485 TI - [Advances in research on monoclonal antibody agents for cancer therapy]. AB - Research and development of antitumor monoclonal antibody agents have shown substantial progress. Monoclonal antibodies (McAb) or their immunoconjugates can be used as therapeutic agents for cancer treatment. As reported, McAb agents display specific binding to tumor-related targets, exhibit selective cytotoxicity to target cancer cells, and show highly therapeutic effects in animal experiments. Currently, some of the McAb agents have been applied in clinical therapy. Major trends in the study of McAb agents are searching for new relevant molecular targets, humanizing of the antibody, and downsizing of the immunoconjugate molecule. Because McAb is highly specific to related molecular target, there is a great potential to develop highly effective antitumor agents. McAb agents may play an important role in cancer therapy. PMID- 12903486 TI - [Changes of AC/cAMP system and phosphorylation regulation of adenylate cyclase activity in brain regions from morphine-dependent mice]. AB - OBJECTIVE: To further understand the changes of AC/cAMP system in the brain regions from morphine-dependent mice. METHODS: By inducing morphine dependence in mice, we observed changes in AC/cAMP signal system, the phosphorylation regulation of adenylate cyclase (AC) activity in brain regions and effect of protein kinase A (PKA) inhibitor on the development of morphine dependence. RESULTS: (1) In morphine-dependent mice, AC activity, cAMP contents, and cytosolic PKA activity in striatum, hippocampus, and cerebral cortex were significantly higher than those of control. But there were no similar changes in cerebellum, and PKA inhibitor injected intracerebroventricular 15 min prior to morphine injection could inhibit the changes of AC activity; (2) These changes described above were not observed in mice treated with naloxone 30 min prior to daily morphine injection; (3) In striatum and cerebral cortex of morphine dependent mice, level of AC phosphorylation in vitro was apparently higher as compared to control group. It indicated that the level of AC phosphorylation in vivo was decreased in morphine-dependent mice; (4) PKA inhibitor was given to mice intracerebroventricular (i.c.v.) 15 min. prior to daily morphine injection could prevent the development of morphine dependence in mice. CONCLUSIONS: The up regulation of AC/cAMP-PKA signal system in some brain regions occurred in the state of morphine dependence and the effect was mediated via specific activation of opiate receptors. Furthermore, that PKA affecting AC phosphorylated state and leading to the increase of AC activity suggest that the increase of PKA activities during chronic morphine treatment could lead to the positive feedback regulation in AC/cAMP system and further potentiate the AC/cAMP system, and these may be one of important mechanisms by which chronic opiate induce dependence in target neurons. PMID- 12903487 TI - [Effects of oxidized LDL, hypoxia, and 5-hydroxytryptamine on 5-HT2A receptor and intracellular free Ca2+ in cultured vascular smooth muscle cells]. AB - OBJECTIVE: To investigate the mechanisms of this hyperreactivity, the effects of oxLDL, hypoxia, and 5-HT (the relative factors inducing arteriosclerosis) on 5 HT2A receptor, receptor gene expression and intracellular free Ca2+ were observed in cultured rat aortic smooth muscle cells (ASMC). METHODS: ASMC underwent 50 micrograms/ml oxLDL 8 hours, 2% hypoxia 24 and 48 hours, 10 mumol/L 5-HT 30 min. 5-HT2A receptor, 5-HT2A receptor mRNA and [Ca2+]i of ASMC were estimated by radiobinding assay, RT-PCR and Southern blot method, and confocal laser scanning mode separately. RESULTS: After oxLDL, hypoxia, 5-HT pretreatment, 5-HT2A receptor of ASMC showed significant up-regulation. Both 5-HT2A receptor mRNA expression and 5-HT-induced [Ca2+]i of ASMC increased markedly after oxLDL, hypoxia pretreatment. CONCLUSIONS: 5-HT2A receptor up-regulation and cell [Ca2+]i increase of ASMC induced by oxLDL, hypoxia, and 5-HT may play a role in vascular hyperreactivity induced by 5-HT in atherosclerosis arteries. PMID- 12903488 TI - [Identification of early irreversible damage area in a rat model of cerebral ischemia and reperfusion]. AB - OBJECTIVES: To observe the early neuron ischemic damage in focal cerebral ischemia/reperfusion with histostaining methods of argyrophil III (AG III), Toludine blue(TB), and H&E, and to make out the 'separating line' between the areas of reversible and irreversible early ischemic damage. METHODS: Forty-two male Wistar rats were randomly divided into the following groups: pseudo surgical, blank-control, O2R0(occluded for 2 hours and reperfused for 0 hour), O2R0.5, O2R2, O2R4, O2R24. There were 6 rats in each group. Rats in experimental groups were suffered focal cerebral ischemia/reperfusion through a nylon suture method. After a special processor for tissue manage, the brain were coronal sectioned and stained with H&E, TB, and AG III. The area where dark neurons dwell in (ischemic core) were calculated with image analysis system. RESULTS: The success rate of ischemic model for this experiment is 90%. After being stained with argyrophil III method, normal neurons appear yellow or pale brown, which is hardly distinguished from the pale brown background. The ischemic neuron stained black, and has collapsed body and "corkscrew-like" axon or dentries, which were broken to some extent. The neuropil in the dark neurons dwelt area appears gray or pale black, which is apparently different from the pale brown neighborhood area. The distribution of dark neurons in cortex varies according to different layers, and has a character of columnar form. The dark neurons present as early as 2 hours ischemia without reperfusion with AG III method. CONCLUSIONS: AG III stain could selectively display early ischemic neurons, the area dwelt by dark neurons represent early ischemic core. Dark neuron is possibly the irreversibly damaged neuron. Identification of dark neurons could be helpful in the discrimination between early ischemic center and penumbra. PMID- 12903489 TI - [High level expression of foreign gene via multiple joined operons and a new concept on the restricted constant of total amount of plasmid DNA per cell of Escherichia coli]. AB - OBJECTIVE: To examine the feasibility of linking operons in tandem to enhance expression of heterologous genes in Escherichia coli and to clarify the potential control mechanism of the total plasmid DNA amount in each host cell. METHODS: Two series of expression plasmids, CW11 and CW12, containing 1 to 4 and 1 to 3 heterologous gene operon(s), respectively, were constructed. The molecular size of the CW11 series varied from 5.47 kb to 12.26 kb in 2.25 kb increments. The CW12 series varied from 5.40 kb to 9.72 kb in 2.16 kb increments. Protein expression was analyzed via SDS-PAGE and laser scanning. Plasmid copy number was determined by incorporation with 3H-TdR([3H]-thymidine). RESULTS: No influence of the tandem-joined operons on host growth and plasmid stability was observed. Upon induction, the desired protein accumulated in the CW11 series were 46.0%, 54.8%, 56.1%, and 60.1% of total cell protein. In the CW12 series, the yields were 33.5%, 44.0%, and 47.1% of total cell protein. With increasing size, however, plasmid copy number decreased. Further calculation showed that the total amount of plasmid DNA per cell is restricted to some extent. There seems exists a mechanism to control the maximum level of total plasmid DNA amount in each host cell. CONCLUSIONS: Increasing the target gene dosage by tandem linking of operons enhanced the expression level of a desired protein. Although the size (kb) and the copy number of each plasmid are negatively interrelated, it seems that as for certain plasmids, their total DNA amount per cell is a restricted constant for a specific E. coli strain under identical incubation condition. PMID- 12903490 TI - [Association of HLA-DRB genes with Vogt-Koyanagi-Harada syndrome in a Chinese Han population]. AB - OBJECTIVE: To investigate the molecular genetic background of VKH syndrome in Han Chinese by HLA-DRB genotyping. METHODS: HLA-DRB genotyping was performed by PCR SSOP method, then DR4 alleles were analyzed by SSOP and RFLP methods. RESULTS: The frequency of DRB1 * 0405 was significantly higher in VKH patients than that in normal controls (80.4% vs 8.1%, Pc < 10(-7), RR = 46.7), so was the frequency of DRB1 * 15 in DRB1 * 0405 negative individuals (Pc < 0.05, RR = 6.0). On the other hand, the frequencies of DRB1 * 14 and DRB1 * 08 were significantly lower in VKH patients than that in normal controls. Comparing the clinical features between DRB1 * 0405 positive and negative VKH patients, no significant differences were found. CONCLUSIONS: (1) DRB1 * 0405 and DRB1 * 15 are closely associated with the susceptibility to VKH syndrome, DRB1 * 0405 may be the major susceptible gene and DRB1 * 15 may be the minor; (2) DRB1 * 14 and DRB1 * 08 are negatively associated with the susceptibility to VKH syndrome, suggesting that they may be the resistant genes; (3) DRB1 * 0405 is not related to the clinical features, incidence of ocular complications as well as visual prognosis. PMID- 12903491 TI - [Diagnosis and management of gestational trophoblastic tumor with liver metastases]. AB - OBJECTIVE: To evaluate management options and clinical prognosis for women presenting gestational trophoblastic tumor (GTT) with liver metastases. METHODS: We retrospectively analyzed sixteen GTT patients with liver metastases treated in the Peking Union Medical College (PUMC) hospital from 1985 to 1998. All were treated with 5-FU combined chemotherapy or EMA/CO regimen. Eight of them received hepatic artery infusion chemotherapy. Treatment outcome was measured by beta subunit human chorionic gonadotropin assay (beta-hCG) and by imaging studies which included ultrasound and computerized tomography. RESULTS: Of 16 cases, 4 achieved complete remission; biochemical remission was obtained in 2 patients who were alive with residual tumor; 10 patients died of the disease. The overall survival rate was 37.5% (6/16). CONCLUSIONS: The outcome for women presenting with liver metastases from GTT is poor. Early diagnosis and prompt initiation with vigorous multi-agent chemotherapy given through multi-routes are emphasized for improving the treatment outcome. PMID- 12903492 TI - [Detection of DNA variation after space flight in Datura innoxia by random amplified polymorphic DNA markers]. AB - OBJECTIVE: To substantiate the effect of space environment on medicinal plants, the seeds of Datura innoxia were set up in retrievable satellites. METHODS: After returning to earth, the DNA variation of different groups was detected by random amplified polymorphic DNA (RAPD) markers. RESULTS: From a pre-screening of 65 primers with 20 oligonucleotide lengths, 15 primers that produced distinct profiles for each DNA sample were selected. In contrast with the earth controlled group, 39 polymorphic bands were produced in weightless group, and its degree of gene polymorphism was 23.1%; 45 polymophic bands were produced in hit group, and its degree of gene polymorphism was 24.4%. The polymorphic bands ranged approximately from 200 to 1990 bp. CONCLUSIONS: These results indicate that weightlessness induce DNA variation to some extents in Datura innoxia, while the compound effect of weightlessness and high energy heavy ions is more notable than the effect of single weightlessness. PMID- 12903493 TI - [Investigation of connective tissue growth factor's gene cloning]. AB - OBJECTIVE: The human connective tissue growth factor(CTGF), as the responsive product of immediate early gene, was cloned for the investigation of cellular primary gene responded and healing repairing. METHODS: RT-PCR was performed with the mRNA of HUVE cell stimulated, the three clones of CTGF, via mutational modified, were ligated to a complete gene that identified with sequencing. RESULTS: The sequencing indicated that this CTGF gene contains complete coding region and the 3'-noncoding region which is difference with the reported sequence. CONCLUSION: A new human CTGF gene was cloned. PMID- 12903494 TI - [Expression and glycosylation of rotavirus strain SA11 VP4 protein in a recombinant adenovirus]. AB - OBJECTIVE: A modified VP4 gene of rotavirus SA11 strain was expressed by a recombinant human adenovirus. METHODS: A whole VP4 gene was obtained with PCR and induced the signal peptide at the gene N terminal. The chimera gene was cloned into pCMV plasmid that consist of human cytomagolovirus promoter and then cloned the gene to transfer human adenovirus type 5 vector. Homologues recombinant was performed by co-transfection to 293 cell line with recombinant plasmid and viral genome using CaPO4 precipitation. RESULTS: VP4 gene is 2,362 base pair in length mutation was not found in whole VP4 gene sequence. Expressed product in recombinant adnovirus was confirmed to be specific and more antigenicity by indirect immunofluorescence assay. Both Western blot and immunoprecipitation assays showed that expressed protein molecular weight was higher than wild type VP4 protein and that modified product was corresponding to a glycosylation of VP4 protein. CONCLUSIONS: It may be a effective method to modify interested gene for enhancing stability, antigenicity and immunogenicity of expressive product. PMID- 12903495 TI - [Result of p53, ki-67 protein expression in cholangiocarcinoma with in situ hybridization and immunohistochemistry methods]. AB - OBJECTIVE: To clarify the clinical significance of P53, ki-67, and the relationship between p53 gene expression and ki-67 labeling index. METHODS: In this study, 30 cases of human cholangiocarcinoma, paired with normal bile duct tissue adjacent to the tumor, were included. In situ hybridization (ISH) was used to detect the p53 gene expression. Immunohistochemistry (IHC) was applied to analyze p53 gene mutation and ki-67 labelling index. RESULTS: ki-67 and P53 positivity were detected in 96.7% (29/30) and 53.3% (16/30) of carcinoma specimens respectively by IHC, neither in peripheral normal tissue. p53 positivity was related to local lymph node metastasis. p53 positivity of cholangiocarcinoma in clinical stage III was significantly higher than that in clinical stage I, II. The expression of ki-67 protein was correlated with local lymph node metastasis and clinical stage. In lymph node metastasis group and clinical stage III group, ki-67 labeling index was significantly higher than that in non-lymph node metastasis group as well as in early stage group. We also found ki-67 labeling index of moderately or poorly differentiated cholangiocarcinoma was significantly higher than that of well differentiated carcinoma. Comparing survival time, the median survival time of patients whose bile duct tumors expressed no P53 protein or with lower ki-67 index was longer than that of patients whose tumors expressed P53 protein, or with higher ki-67 labeling index. CONCLUSIONS: The results indicate that p53 positivity may be correlated with tumor development, growing activity. They also indicate that ki-67 labeling index reflects the degree of tumor cell proliferation. This observation suggests that ki-67 labeling index may be a useful predictor of tumor prognosis. PMID- 12903497 TI - [Arsenic trioxide induced human esophageal cancer Ec109 cell apoptosis with downregulation of c-myc gene expression]. AB - OBJECTIVE: To study effect of downregulation of c-myc gene expression with As2O3 on inducing Ec109 cell apoptosis. METHODS: MTT reduction assay, flowcytometric analysis, DNA gel electrophoresis, Western blot and in situ cell death detection (TUNEL). RESULTS: The viability of the cells treated with As2O3 was decreased. The cells treated with As2O3 showed a typical apoptotic morphology and hypodiploid peak before G1 phase. DNA of treated Ec109 cells appeared a ladder pattern characteristic of apoptosis. TUNEL detection analysis also revealed the DNA fragmentation and As2O3 decreased c-myc gene expression was observed. CONCLUSIONS: As2O3 can induce Ec109 cell apoptosis with downregulation of c-myc gene expression. PMID- 12903496 TI - [Bone mineral density and exercises: a cross-sectional study on Chinese athletes]. AB - OBJECTIVES: To evaluate the effect of exercise to osteoporosis by bone mineral density (BMD) measurement of athelets comparing normal individuals in general population. METHODS: BMD of radium, lumber spine, and femoral neck were measured by single photon absorptiometry (SPA), quantitative CT (QCT), and dual X-ray absorptiometry (DXA) respectively in athletes (n = 162, male 79, female 83) and age matched non-athletes normal population (n = 204, male 91, female 113) in Beijing. RESULTS: BMD of all sites in all age groups of both male and female athletes are significantly higher comparing with that in non-athletic population. This predominance in athletes is even more distinctive in peak bone mass. Peak bone mass of male athletes is significantly higher than that of female athletes. Bone loss with age is less apparent in athletes than in control. However, there is an accelerated decline of BMD in lumber spine and femoral neck in 30-39 year age group in both male and female athletes, which may be due to the wanting of physical exercise. CONCLUSIONS: Long term regular proper exercise started in adolescence may play a very important role in the prevention of osteoporosis by improving peak bone mass and decreasing bone loss. PMID- 12903498 TI - [Characteristic immunodeficiency syndrome of rapid fatal type of simian immunodeficiency virus infected monkeys]. AB - OBJECTIVE: To observe the characteristic immunodeficiency syndrome of the rapid fatal type of simian immunodeficiency virus (SIV) infected monkeys. METHODS: Eighty rhesus monkeys and 4 cynomolgus monkeys were intravenously inoculated with SIVmac or SIVmac251. The virus isolation and viral titer, estimation by indirect immunofluroresence and viral antibody were determined periodically from monkeys' plasma; lymph node biopsies were performed for pathohistological examination. RESULTS: Twelve out of 84 macaque (14.2%) died of rapid progressive type after inoculation of SIVmac and SIVmac251 in the duration 3 to 4 months. Dying monkeys showed persistent high viremia and low level titre antibody. Eight of 10 pathohistological changes showed severe depletion of lymphoid tissue in spleen and lymph nodes, there were remarkable immunodeficiency with opportunity infection. The other two monkeys appeared moderate lymphoid tissue deletion and hyperplasia without opportunity infections. The survived monkeys' (72/84) lymph nodes biopsies revealed hypoplasia of lymphoid tissue. CONCLUSIONS: The characteristic immunodeficiency syndrome of rapid fatal type of simian immunodeficiency virus infected monkeys could be made with persistent high viremia, low level antibody, severe lymphoid tissue deletion in lymph nodes and spleen, as well as complicated opportunity infections. PMID- 12903499 TI - [Prediction of peritoneal fluid pancreatic amylase to histopathological change and prognosis of acute experimental necrotic pancreatitis of monkey]. AB - OBJECTIVE: To observe the significance of peritoneal fluid pancreatic amylase in predicting the degree of histopathological change and prognosis of acute necrotic pancreatitis (ANP) of monkey. METHODS: Injection of the mixture of bile and trypsin into the pancreatic duct set up ANP monkey model. RESULTS: The severity of ANP depended on the injection rate and volume, in correlation to the peritoneal fluid pancreatic amylase (PF Amylase) in the early stage of ANP. With the rate (3-13 ml/min) and volume (0.1-1.0 ml/kg) of injection increase, the PF Amylase in the early stage of the ANP significantly went up. Meanwhile, the survival time was negatively correlated to the PF Amylase in the early stage (0.5 hour) of ANP (t = -9.06 InX + 129.85, r = -0.88, P < 0.01). CONCLUSIONS: The PF Amylase in the early stage of ANP can effectively predict the severity of ANP and prognosis. PMID- 12903500 TI - [Prevention and treatment of bladder tumors recurrence with furtulon]. AB - OBJECTIVE: To inquire into the value of oral furtulon therapy for bladder tumors. METHODS: 103 patients were randomized into four groups and have been undergoing a long term follow-ups. RESULTS: Recurrent rate of oral furtulon therapy for bladder tumors (8.8%) and instillation of mitomycin C for preventing the recurrence of bladder tumors (12.5%) as compared to recurrent rate of control group(40%), which was statistically significant(P < 0.01). Overall, 4 patients (33.3%) had an effective response with furtulon. CONCLUSIONS: Furtulon has more effective for treatment and prophylaxis of bladder tumors. PMID- 12903501 TI - [High dose intravenous immunoglobulins in treating systemic lupus erythematosus associated thrombocytopenia]. AB - OBJECTIVE: To study the effect of high dose intravenous immunoglobulins(IvIg) in systemic lupus erythematosus (SLE)-associated thrombocytopenia. METHODS: A case control study was undertaken to assess the effectiveness of different therapeutic regimens (oral prednisone, IvIg, bolus methylprednisolone(MP), and IvIg plus bolus MP) in improving SLE-associated thrombocytopenia. RESULTS: For SLE associated thrombocytopenia, IvIg, bolus MP, and IvIg plus bolus IvIg significantly shorten the time to reach a peak platelet count, which was (6.2 +/- 4.4) days, (7.3 +/- 4.9) days, and (3.8 +/- 2.4) days respectively in comparison with oral prednisone, which was (31.0 +/- 17.8) days (P< 0.01); if high dose IvIg and MP in combination, the time was also shorter than bolus MP alone (P < 0.05). The elevation of platelet count in groups receiving IvIg, bolus MP, and IvIg plus bolus MP were significantly higher than as in group receiving oral prednisone therapy(P < 0.05). CONCLUSIONS: High dose IvIg is a useful therapy in improving SLE-associated thrombocytopenia. PMID- 12903503 TI - [Prevention and management of complications with laparoscopy cholecystectomy]. AB - OBJECTIVE: To prevent the complication of laparoscopy cholecystectomy (LC). METHODS: This text analysed 1,200 cases retrospectively with LC from Feb. 1993 to May. 1998. RESULTS: Three cases with biliferous duct injury, two cases with bile leakage from the stamp of cystic duct, two cases with gallbladder artery injury. CONCLUSIONS: Strict preoperative examination and standard operative procedure can reduce markedly complication of LC. PMID- 12903502 TI - [A simplified six-item checklist for screening of fragile X syndrome]. AB - OBJECTIVE: To investigate whether a simplified six-item checklist could be developed to improve the screening fragile X syndrome test result. METHODS: Nine clinical characteristics were selected from patient records of 190 male and 18 female pediatrics for fragile X screening test were analyzed. The characteristics included mental retardation, family history of mental retardation, elongated face, large or prominent ears, attention deficit hyperactivity disorder, Autistic like behavior, simian crease, macroorchidism, and hyperextensible joints. RESULTS: Seven cases were diagnosed with fragile X syndrome by Southern analysis on PCR product. Among the nine characteristics, simian crease, macroorchidism, and hyperextensible joints were eliminated, because of low frequency and statistical insignificance. Using remaining six-item clinical checklist, if a score of 6 or more was used as the criteria for screening fragile X test, about 60% of this test in our cases could have been eliminated clinically without missing any positive cases. Thereby the proportion of case with positive results improved 8.8%. CONCLUSIONS: With our simplified six-item clinical checklist, 60% of testing could have been eliminated clinically, thereby improving the effectiveness of fragile X screening test and promoting the proportion of cases with positive results in two groups. PMID- 12903504 TI - [Effect of titanium substructure on color of porcelain-fused-to-titanium restoration]. AB - OBJECTIVE: To study the effect of titanium substructure on color of porcelain fused-to-metal restoration. METHODS: Standard samples of Dentsply DETREY-Carat and Tibond porcelain fused-to-metal restorations were made. Color Parameters were tested by cs-5 spectrophotometer. Statistic analysis was done and the color differences were calculated. RESULTS: Most of color parameters were found to be statistically significant at the P < or = 0.05 level at substructure layer. But no statistical significance existed after putting opaque and dentin porcelain. The values of color difference between two groups at opaque and dentin porcelain layers were all smaller than 1.5. CONCLUSIONS: Titanium substructure had no visually apparent effect on final color of porcelain-fused-to-metal restoration. There was no perceived color difference between corresponding shade of two Dentsply porcelain systems (DETREY-Carat and Tibond). PMID- 12903505 TI - [Establishment of ELISA for measuring phenytoin in serum]. AB - OBJECTIVE: To develop a solid phase, competitive enzyme immunoassay for the measurement of serum phenytoin. METHODS: The chemical modified phenytoin combined with human serum albumin and then conjugated with horseradish peroxidase (HRP) to produce the enzyme labeled phenytoin (DPH-HSA-HRP). The anti-phenytoin antibody was prepared in this lab. RESULTS: The working range and sensitivity of this method were 2.9-30 micrograms/ml and 2.87 micrograms/ml respectively. The intra assay coefficient of variation (CV) was 3.3%-10.2% (n = 15) and inter-assay CV was 5.1%-13.2%. The recovery of this method was 90%-96%. No significant interference was observed with phenobarbital, primidone, carbamazepine, and valproic acid. The new assay method was compared with HPLC method. A linear regression analysis yielded a slope of 1.03, an intercept 1.38 and a correlation coefficient 0.97. CONCLUSIONS: This solid phase enzyme immunoassay for serum phenytoin appears to be simple, precise, and accurate. It may be readily adopted in clinical laboratory for therapeutic monitoring of phenytoin level in serum. PMID- 12903506 TI - [The past, current status and future of general practice and community health service in China]. AB - Some basic concepts of general practice and community health service are introduced briefly in this paper, as well as their past, current status and future. Issues existed in general practice and community health service at present are also discussed. In fact, general practice and community health service have existed in China for a long time since 1920s. The urgency and possibility for development of general practice and community health service in China are analyzed. Suggestions for three issues relevant to their development are put forward. PMID- 12903507 TI - [Introduction of the high throughput screening system]. AB - In this article, we introduce the system of HTS (high throughput screening). Its role in new drug study and current development are described. The relationship between research achievements of genome study and new type screening model of new drugs is emphasized. The personal opinions of current problems about HTS study in China are raised. PMID- 12903508 TI - [Study on the role of superoxide anion in promoting proliferation and transformation of rat liver oval cell line WB-E344]. AB - OBJECTIVE: To study the role of superoxide anion (O2.-) in promoting proliferation and transformation of rat liver oval cell strain WB-F344. METHODS: WB-F344 cells cultured were stimulated directly by O2.- generated by interaction of xanthine with xanthine oxidase (X-XO). The effect of O2.- in promoting proliferation of WB cells was investigated by using MTT colorimetric analysis, 3H Tdr incorporation liquid scintillation counter and 3H-Tdr incorporation autoradiography. WB cells initiated with N-methyl-N'-nitro-N nitrosoguanidine(MNNG) were promoted by stimulating continuously with O2.- of low concentration (X: 100 mumol/L, XO: 0.2 mU/ml). The transformation effect was tested by morphologic observation, karyotype analysis and anchorage-independent growth assay. RESULTS: Proliferation of WB cells was induced obviously by O2.- of low concentration for only one time. WB cells initiated with MNNG were transformed by action with O2.- of low concentration continuously for 15 d and typical morphologic character of transformed cells was observed. In karyotype analysis the cells chromosome number changed and the frequency of structure aberration raised dramatically. Also the transformed cells could form clone on self-solid culture medium. CONCLUSIONS: The biological effect of O2.- was related closely with its dose; The effects of low concentration in promoting proliferation and transformation of liver oval cells indicate its important role in hepatocarcinogenesis and antioxidation was able to provide a new clue in prevention and cure of hepatoma. PMID- 12903509 TI - [Expression and deletion analysis of EcoR II endonuclease and methylase gene]. AB - OBJECTIVE: To clone complete EcoR II restriction endonuclease gene (ecoR II R) and methyl-transferase (ecoR II M) gene into one vector and to analyzing the expression of the whole system. METHODS: Unidirective deletion subclones constructed with Exo III, ecoR II R/M genes were preliminarily located in the cloned fragments according to the enzyme activities of each subclone, exact deletion sites were determined by sequencing, and transcriptional start sites were mapped by S1 mapping. RESULTS: The DNA fragment which was cloned into pBluescript SK+ contained the complete ecoR II R gene and ecoR II M gene, there are two transcriptional start sites in ecoR II R gene, 132 bp to 458 bp from 3' and of ecoR II R gene are indispensable to enzyme activities and deletion of 202 bp from 3' end of ecoR II M gene made it lose the capability to resist specific cut of EcoR II R enzyme, deletion of coding region and flanking sequence of one gene did not affect the expression of the other gene, the recombinant only containing ecoR II R gene appeared to be lethal to dcm + host. CONCLUSIONS: ecoR II M gene closely linking to ecoR II R gene was very important for the existence of the R-M system in process of evolution, but the key to control EcoR II R enzyme acted later than EcoR II M enzyme did not exist in transcriptional level. PMID- 12903510 TI - [Epidemiological characteristics of Guillain-Barre syndrome in urban and rural areas in Beijing and Hebei, China]. AB - OBJECTIVE: To investigate epidemiological and clinical patterns of Guillain-Barre syndrome (GBS) in urban and rural areas in Beijing municipality and Hebei province, China. METHODS: We investigated GBS incidence using a strengthened case surveillance and an active case ascertainment in 2 counties and 4 districts of Beijing municipality and 3 counties of Hebei province during 1993 to 1994. RESULTS: On the basis of the diagnostic criteria of NINCDS, 54 patients were identified. The age-adjusted incidence rates per 100,000 population for GBS were 0.9 in rural areas, and 0.8 in urban areas. A peak agespecific incidence showed in adults aged 50 to 59 years. A higher incidence appeared to occur in the spring and summer for rural residents, but not significant for urban population. In comparing course of GBS in rural and urban areas, there were differences in mean days from beginning of neurological symptom to maximal weakness (4.3 vs 7.6 days), and from symptom onset to beginning of recession (11.8 vs 17.5 days). There were preceding events in 72% patients, most frequently in respiratory infection, sensory disturbance in 70.9%, and respiratory assistance in 7.6%. The outcome was compatible with other reports; with complete recovery at 12 months in 79.2% and minimal residua in 20.8% for those alive and with casefatality rate in 7.4%. In addition, a follow-up study on electrophysiological features in 19(90.5%) patients from two counties of Beijing showed the demyelinating lesion (89.5%) over the axonal lesion (52.6%) of motor and/or sensory nerves. CONCLUSIONS: The epidemiological and clinical characteristics of GBS were similar to that reported in other countries. Demyelinating GBS was the main pattern in present population-based study. PMID- 12903511 TI - [Association of microtubule promoting and binding activity of tau with its phosphorylation sites]. AB - OBJECTIVE: To explore the association between the abnormal phosphorylation sites found in Alzheimer disease (AD) tau and the inhibition of its biological activity. METHODS: Ultracentrifugation, chromatography, manual Edman degradation and autosequence techniques were used to prepare and phosphorylate human recombinant tau, isolate and purify 32P tau peptides and determine phosphorylation sites. RESULTS: (1) Phosphorylation of tau by casein kinase-1 (CK 1), cyclic AMP-dependent protein kinase (PKA) and glycogen synthetase kinase-3 (GSK-3) differentially inhibited its biological activity, and the inhibition of this activity of tau by GSK-3 was significantly increased if tau was prephosphorylated by CK-1 or PKA. The most potent inhibition was seen by a combined phosphorylation of tau with PKA and GSK-3. (2) The treatment of tau by PKA and GSK-3 combination induced phosphorylation of tau at Ser-195, Ser-198, Ser 199, Ser-202, Thr-205, Thr-231, Ser-235, Ser-262, Ser-356, Ser-404, whereas only Thr-181, Ser-184, Ser-262, Ser-356 and Ser-400 were phosphorylated by GSK-3 alone under the same conditions. Among the above-mentioned phosphorylation sites, Ser 198, Ser-199, Ser-202, Thr-205, Thr-231, Ser-235, Ser-262, Ser-400 and Ser-404 were seen in Alzheimer tau. The phosphorylation of Ser-262 only slightly inhibited its biological activity, and Ser-198, Ser-199, Ser-202, Thr-231, Ser 235, Ser-400 and Ser-404 also presented in fetal tau which was highly active, suggesting that Thr-205 was the unique site which both caused the potent inhibition of biological activity and specifically presented in AD abnormally phosphorylated tau. CONCLUSIONS: Phosphorylation of Thr-205 might play a key role in tau pathology in AD. PMID- 12903512 TI - [Arsenic trioxide induces human tumor cell apoptosis and G2 + M arrest whereas causes HPV16 DNA immortalized cervical epithelial cells G1 block]. AB - OBJECTIVE: To investigate whether As2O3 induces tumor cell apoptosis and cell cycle arrest. METHODS: DNA agarose gel electrophoresis, flowcytometry analysis, RT-PCR, and Western blot. RESULTS: As2O3 induced G2 + M arrest of tumor cells such as GLC-82, MGC-803, SGC-7901, Eca109 and HeLa cells before inducing apoptosis and decreased c-myc gene expression, whereas caused G1 block of HPV16 DNA immortalized cervical epithelial HCE16/3 cells and did not affect c-myc expression. CONCLUSIONS: As2O3 caused apoptosis was related with cell cycle arrest. However, the phase of arrest of tumor cells was different from the pre tumor cell HCE16/3 cells, which could be related to the change of c-myc gene expression. PMID- 12903513 TI - [Studies on the stable integration and expression of IL-2R alpha gene in CHO cells and it's applications]. AB - OBJECTIVES: To establish a novel bioassay method for quantitative analysis of human IL-2 based on the specific binding of Interleukin-2 receptor alpha subunit (IL-2R alpha) with IL-2. METHODS: Southern blot hybridization was first used to detect the stability of integration of recombinant secretive IL-2R alpha (rsIL-2R alpha) gene into the genome of highly expressed cell line reported elsewhere; the apparent Mr of the rsIL-2R alpha was then determined by using Western blotting; finally, a receptor-antibody sandwich ELISA method has been established for quantitative analysis of IL-2. RESULTS: (1) Stable integration of rsIL-2R alpha gene into the genome of #17 CHO cell line has been identified; (2) the apparent Mr of rsIL-2R alpha was approximately 40,000; (3) linear range of the standard curve obtained from the receptor-based ELISA fell between 31.25-500 U of IL-2 (r = 0.9995). The slop of the standard curve decreased significantly when IL-2 was pre-incubated with goat anti-IL-2 antibody IgG (P < 0.01). CONCLUSIONS: An IL-2R alpha-based IL-2 ELISA has been established for laboratory and clinical use with advantages of accuracy, specificity and simplicity over other conventional bioassays for IL-2 detection. PMID- 12903514 TI - [Effects of high dose of dynorphin on NMDA receptor and NOS activities in spinal cord of rats]. AB - OBJECTIVE: To elucidate the effects of N-methyl-D-aspartate(NMDA) receptor and nitric oxide synthase (NOS) activity in dynorphin (Dyn)-induced spinal cord injury. METHODS: The NMDA receptor activity was measured by radio-ligand of 3H MK801. The constitutive and inducible NOS (cNOS and iNOS) activities were assayed by 3H-arginine conversion. RESULTS: In ventral samples, both 3H-MK801 binding and cNOS activity increased at 0.5 h and persisted for 48 h while iNOS activity enhanced at 4 h after intratheacal injection (i.t.) Dyn A(1-17) at dose of 20 nmol/L. However, the 3H-MK801 binding activity reduced significantly from 4 h to 24 h and cNOS activity did not change at the same time in dorsal samples. 7 nitroindozol (7-NI) and aminoguanidine (AG) inhibited the effects of Dyn A(1-17) (20 nmol/L) on 3H-MK801 binding and NOS activities in ventral samples. N-nitro-L arginine methyl ester (L-NAME) did not affect the elevation of Dyn A(1-17) on NOS activities but caused 3H-MK801 binding activity reduction in ventral samples. CONCLUSIONS: NMDA-NOS pathway might play important role in Dyn spinal neurotoxicity. NOS inhibitors and Dyn might produce cooperative down-regulation on the function of NMDA-NOS pathway in dorsal cord. PMID- 12903515 TI - [Cytosine diaminase/5-flurocytosine mediated inhibiting effect on the growth of human pancreatic carcinoma cells]. AB - OBJECTIVE: To study the inhibition effect of E. coli cytosine diaminase (CD)/5 flurocytosine (5-FC) system on the growth of human pancreatic carcinoma cell line cells. METHODS: Recombinant retroviral vector expressing CD genes was transduced into pancreatic carcinoma cell line cells. In vitro and in vivo prodrug sensitivity assays were carried out, including: (1) detection of growth and colony forming inhibition rate of transduced cells in the presence of 5-FC; (2) observation of bystander effect by MTT method; (3) observation of the growth inhibition effect of 5-FC on transduced cell xenografts in nude mice. RESULTS: The growth and colony forming inhibition rate of the transduced cells in the presence of 5-FC were 80.0% and 79.0%, which were obviously higher than 4.8% and 5.0% (P < 0.01) of nontransduced cells. Mixed cells containing only 10% of transduced cells showed 50.7% reduction of proliferation (bystander effect). The growth of transduced cells xenografts could be totally regressed by 5-FC. CONCLUSIONS: CD/5-FC system is a potential gene therapy strategy for pancreatic carcinoma. PMID- 12903516 TI - [The role of 5'proximal fragment of POMC gene in the regulation of transcription in AtT20 cells]. AB - OBJECTIVE: Identification of the vital regions in 5'-flanking sequence of POMC gene in order to determine the characteristics of POMC gene promoter and provide the basis for stadying the effects of multiple hormones and other factors in the POMC gene expression. METHODS: Construction of four kinds of plasmid recombined by rat POMC gene promoter regions and eukarytic expression reporter gene vector pG12-Basic. Each kind of them was identified by restriction maps analysis and DNA sequencing. All recombinant plasmids were transfected into AtT20 cells respectively by using a lipofectamine-based method. Luciferase activity in each cell-lytate was measured by monolight 2010 luminometer. RESULTS: Four recombinant plasmids were constructed, including pGL2-POMC480 (rPOMC -480/+63), pGL2-POMC323 (rPOMC -323/+63), pGL2-POMC165 (rPOMC -165/+63). pGL2-POMC34 (rPOMC -34/+63) Among them, PGL2-POMC34 contains the core promoter which consist of TATA box at 31/-25 bp and transcription start site at +1 bp of POMC gene. It showed that, comparing with the pGL2-POMC34, the relative luciferase activity of pGL2-POMC165 was 2.1 +/- 0.3, pGL2-POMC323 was 3.3 +/- 0.3, pGL2-POMC480 was 3.7 +/- 0.5, and the positive control pGL2-promoter was 4.8 +/- 0.8. CONCLUSIONS: (1) A model system to study the direct effects of various ACTH secretagogues on the dynamics of the POMC gene promoter activity in vitro was established. (2) The studying of POMC promoter basal transcriptional activities identified that the POMC gene has a weak core promoter. The proximal sequence elements were concentrated in the fragment of -34/-165 bp, and the transcriptional activities need POMC 5' promoter -480 bp or a longer region. PMID- 12903517 TI - [Expression of proteolytic enzymes of metastatic tumor cells and the effects of some influencing factors]. AB - OBJECTIVE: To elucidate the types of proteolytic enzymes the dendritic cell sarcoma (DCS) cell expressed in vitro and the effects of modulating factors. METHODS: The proteolytic spots of DCS cells were examined, with ten different proteinase inhibitors, various antibodies and matrix. RESULTS: Aprotinin, EDTA Na2 and pepstatin could inhibit the proteolysis of DCS cells respectively. Anti ubiquitin antibody and anti-DCS antibody (1:200-1:100) showed no obvious inhibitory effects. DCS cells spread out fairly well but with no proteolysis. CONCLUSIONS: Under the conditions of our experiments the DCS cells had serine proteinase, metalloproteinase and aspartate proteinase activities. Antiubiquitin antibodies and anti-DCS antibodies showed no obvious influence on the proteolysis of DCS cells. DCS cells displayed different proteolysis status on different matrix. PMID- 12903518 TI - [Studies on urinary metabolites of perlolyrine in rats]. AB - OBJECTIVE: To study the metabolism of perlolyrine in rats, which is an active ingredient from the traditional Chinese herb, Ligusticum Wallichii Franch. METHODS: After administration of perlolyrine and deuterated perlolyrine, the rat urines were hydrolyzed with glucuronidase, basified with NaHCO3-Na2CO3, extracted with ethyl ether--iso-propyl alcohol. The organic phases (neutral and basic fractions) were concentrated for trimethylsilyl (TMS) derivatives. The aqueous phase were acidified with sulfuric acid, taken to dryness and extracted with methanol (water soluble acidic fractions) and concentrated for TMS derivatives. The TMS derivatives were determined by gas chromatograph mass spectrometry (GC MS). RESULTS: Perlolyrine and one metabolite were found from the neutral and basic fractions, and two different metabolites were found from the water soluble acidic fractions. CONCLUSIONS: It was proposed that the major metabolic pathways of perlolyrine were that the hydroxylation of perlolyrine and the oxidation of its hydroxylmethyl group. PMID- 12903519 TI - [PGP 9.5 immunohistochemical study on the development of the enteric nervous system of trisomy 16 mouse embryos and their normal littermates]. AB - OBJECTIVE: To study the development of the intestinal innervation of trisomy 16 mouse embryos and their normal littermates from embryonic days 13-18 (ED13-ED18). METHODS: (1) Trisomy 16 mouse embryos used in this study were produced by crossing NMR-1 females with males carrying the two Robertsonian (Rb) translocation chromosomes Rb (11, 16) and Rb (16,17) 8 LubtwLub3. (2) Cytogenetic analysis: accurate ascertainment of trisomy 16 was provided by the demonstration of two Rb metecentric chromosomes and a count of 41 chromosome arms. (3) Rabbit anti-human protein gene product 9.5 antibody was used in PAP and Avidin-biotin methods: RESULTS: At ED13, neither nervous plexuses nor neurons were found in the gut of trisomy 16 mouse embryos and their normal littermates. And in the gut of normal littermates there appear only a scattering neurons with light staining nucleuses at ED14, the irregular network of the myenteric plexus composed of scattering distribution neurons and their processes at ED15 and the regular meshworks of the myenteric plexus with developed ganglia at ED16. However, in trisomy 16 mouse embryos at ED14-ED16, enteric nervous system (ENS) was composed of only some scattering neurons with different distribution density and size. The development of ENS was well in ED17-ED18 normal littermates. The developed myenteric plexus was composed of nervous meshworks with regular distribution. The submucosal plexus had irregularly shaped. The interconnecting strands were frequent visible between the myenteric and submucosal plexuses. In the gut of trisomy 16 mouse embryos the submucosal plexus was absent and the development and differentiation of the myenteric plexus were lower compared with their normal littermates. An important finding was that 5 mm aganglionic bowel was firstly found in the end of colon. In this study, the developed mesentery nervous fibers were persent and innervated in both type animal guts from ED14 to ED18. CONCLUSIONS: It was original report to find the developmental delay of the ENS, absence of submucosal plexus and the aganglionic segment of colon ending in trisomy 16 mice, an animal model for Down syndrome. PMID- 12903520 TI - [Ganciclovir induces apoptosis of rat C6 glioma cell transduced with HSV-tk gene]. AB - OBJECTIVE: To observe whether apoptosis could be induced by ganciclovir (GCV) in the herpes simplex virus-thymidine kinase gene-ganciclovir system (HSV-tk-GCV system) gene therapy for glioma. METHODS: Transduced C6/tk glioma cell as treated group, wild C6 cell as control group, were studied with morphology, gel electrophoresis of DNA fragment analysis, fluoroscopy, and flow cytometric study before or after exposed to GCV, and apoptosis of transduced C6/tk glioma cell was ascertained. RESULTS: When C6/tk cell was treated with GCV at 5 micrograms/ml for 72 hours, cell apoptosis occurred. Typical apoptotic morphological features included cell shrinkage, condensation, and margination of nuclear chromatin were showed by light and electron microscopy, condensed nuclear chromatin and the fragment of nuclei were demonstrated by fluoroscopy, DNA ladder was showed by DNA fragment analysis, apoptotic peak was identified by flow cytometric study. The apoptotic cells accounted for 23% of the cell population. Apoptosis was not observed in control group treated by GCV. CONCLUSIONS: The apoptosis of transduced C6/tk glioma was induced by GCV, which fit in with the basis of HSV-tk GCV gene therapy for glioma. PMID- 12903521 TI - [The summarization of clinical experience of difficult tracheal intubation]. AB - OBJECTIVE: To summarize our experiences and lessons of difficult tracheal intubation for clinical anesthesia reference. METHODS: We had done a retrospective analysis of clinical data on difficult tracheal intubation in 2,825 patients undergoing elective plastic surgery with anesthesia. The main causes of difficult tracheal intubation were the limitations of neck extension (n = 1,169), mouth opening (n = 889), both neck extension and mouth opening (n = 698), and micromaxillary deformity (n = 69). By the Cormack's classification, all the patients had the laryngeal exposure of grade II or more. The tracheal intubations were done under neuroleptanalgesia combined with topical spray of local anesthetic in 439 patients, intravenous anesthesia of sedative drugs and nondepolarizing relaxants of subnormal doses in 629 subjects, and total intravenous or inhaled anesthesia in 1,757 cases, respectively. RESULTS: The difficult tracheal intubations were completed using blind nasal intubations in 142 patients, blind oral intubations with direct laryngoscope in 2,377 patients, oral intubations with fiberoptic stylet rigid laryngoscope in 186 patients, and oral or nasal intubations with flexible fiberoptic bronchoscope in 72 patients. The incidence of successful intubation was 99.7%. The common complication of intubation was airway trauma and its incidence was 19.3% in all the patients. Anesthetic techniques could affect significantly the intubation time and the incidences of complications in the patients with difficult intubation. CONCLUSIONS: By the improvement of anesthetic methods and common intubation techniques, the intubation time and the incidence of complications in the patients with difficult intubations were reduced. PMID- 12903522 TI - [Papanicolaou test in pregnancy]. AB - OBJECTIVE: To investigate the necessity, safety and acceptability of routine Papanicolaou (Pap) smear in pregnant women and cytological characteristics of the smears during pregnancy. METHODS: Pap smear were performed in 954 pregnant women, 1,053 normal controls and 1,908 gynecological out-patients. The cytological preparations were analyzed by using PAPNET method and following Bethesda (TBS NCI) diagnostic criteria. Quality control was carried out by following TBS-NCI standard. RESULTS: Abnormal epithelial change was found in 89 women (89/954, 9.3%) of pregnant group, which was quite similar to normal controls group and gynecological group. Occurrence rate of cervical squamous intraepithelial lesion (SIL) was the lowest in pregnant group (0.94%, 9 in 954 cases), comparing with control group (2.75%) and gynecological group (6.23%). However, pregnant group had the highest infection rate (89.9%), as compared to gynecological group (64.5%), and normal controls group (69.1%) (P < 0.01). CONCLUSIONS: Cervical Pap smear test is necessary, safety and acceptable for routine prenatal tests. Any pregnant women with infections must be treated as nonpregnant one. PMID- 12903523 TI - [Dynamic penumbra in a rat model of focal cerebral ischemia and reperfusion]. AB - OBJECTIVES: To identify the ischemic core and penumbra of variant ischemia and reperfusion period by the dark neuron method, observe them dynamically, and elicit the morphological character of ischemic pneumbra. METHODS: Total 72 male Wistar rats were randomly divided into the following groups: pseudo-surgical, blank-control, O24R0 (sustained ischemia for 24 hours), O4R24(ischemia for 4 hours and reperfused for 24 hours, the following may be deduced by analogy), O2R0, O2R0. 5, O2R2, O2R4, O2R8, O2R24, O2R48, O2R96. There were 6 rats in each group. Rats in experimental groups were suffered focal cerebral ischemia reperfusion through a nylon suture method. After perfusion fixed, the brains were coronal sectioned and stained with HE, TB and acetum plumbi/silver nitrate. Sections were observed with light microscope and electron microscope. The area of ischemic core and penumbra was measured and analyzed with an image analysis system. RESULTS: The penumbra expanded rapidly in a short period after reperfusion, and reach its apex when reperfused for 2 hours (P < 0.05). The penumbra was relatively stable during 4 to 48 hours reperfusion. There were many cell types in ischemic core and penumbra, the proportion of cells changed according to reperfusion time. CONCLUSIONS: Reperfusion could aggravate cerebral edema and make the ischemic penumbra to expand during the first hours. PMID- 12903524 TI - [Changes of plasma nitric oxide and tumor necrosis factor-alpha in lung elastase injury induced inflammatory response of hamster]. AB - OBJECTIVE: To investigate the role of the NO and TNF-alpha in lung elastase injury induced inflammatory response of hamster. METHODS: Lung elastase injury induced inflammatory hamster model was established. NOS activity in airway epithelial cells and lung monocytes/macrophage, as well as the serum content of NO and TNF-alpha were measured respectively. RESULTS: The results indicated that iNOS activity in epithelium and monocytes/macrophages were significantly enhanced. In the early period of inflammation the NO and TNF-alpha content in serum increased obviously and maintained in a high level (P < 0.001). CONCLUSIONS: This Study suggested that both NO and TNF-alpha are important mediators participating in the lung elastase injury-induced inflammatory response. Our data suggested NO might play a regulatory role in the release of TNF-alpha. PMID- 12903526 TI - [Effect of Tripterygium Wilfordii Hook T4 monomer on proliferation and interleukin-6 production of synovial fibroblasts of patients with rheumatoid arthritis]. AB - OBJECTIVE: Tripterygium Wilfordii Hook multi-glycosides T2 has been wildly used in China in treatment of RA. T4 was abstracted from T2 and was reported much more efficient in anti-inflammatory and immune suppression than T2. This study was to investigate the effect Tripterygium Wilfordii Hook T4 monomer on proliferation and interleukin-6 production of synovial fibroblasts of patients with rheumatoid arthritis. METHODS: Synovium was obtained from patients with rheumatoid arthritis undergoing synovectomies or joint replacement. Cultures of synovial fibroblasts were established. After 3 generations, cultured synovial fibroblasts were stimulated with IL-1. Then 1.5 ng/ml, 5 ng/ml and 15 ng/ml T4 were added, and synovial fibroblasts were cultured in the presence of T4 for 48 hours. Cell proliferation was assayed using MTT method. IL-6 level of supernatant was measured by ELISA. RESULTS: Proliferation of synovial fibroblasts was inhibited by T4. The proliferation inhibition effect of T4 was dose dependent and inhibition rate was 5.18%, 10.95% and 21.37%, respectively. And T4 had no effect on IL-6 production by IL-1 stimulated synovial fibroblasts. CONCLUSIONS: T4 might control the disease activity of RA by inhibiting the proliferation of synoviocyte. And T4 might not influence the concentration of IL-6 in synovial fluid, as a central effect, since IL-6 has protective effect on articular cartilage. PMID- 12903525 TI - [Expression of inducible nitric oxide synthase in asthmatic rat lung]. AB - OBJECTIVE: In order to study the role of nitric oxide (NO) during the airway inflammation of asthmatic rat, we investigated expression of inducible nitric oxide synthase (iNOS) in the lung tissue of asthmatic rats. METHODS: Animal model of asthmatic Wistar rats was developed by being sensitized and challenged with ovalbumin. Activities of iNOS were measured by using SP immunohistochemical method. RESULTS: In asthmatic rat group (n = 10), lung tissue showed stronger immunostaining (90%) for iNOS than control group (20%) (P < 0.001). Immunostaining was mainly present in the bronchial epithelium, alveolar epithelium, bronchial smooth muscle, vascular endothelial and smooth muscle cells and inflammatory cells such as macrophages and neutrophils. In glucocorticoid treated group (n = 10), there was down regulation of iNOS immunoreactivity (30% positive) compared with control asthmatic rat group (20%). CONCLUSIONS: The increased expression of iNOS in asthmatic rat lung tissue showed NO playing a critical role in asthmatic inflammation, otherwise expression of iNOS was inhibited by glucocorticoid. PMID- 12903527 TI - [Effect of glazing techniques on color of porcelain-fused-to-titanium restorations]. AB - OBJECTIVE: To study the effect of various glazing techniques on color of porcelain-fused-to-titanium restorations and their surface texture. METHODS: Standard samples of Dentsply Tibond porcelain fused-to-metal restorations were made. Selfglazing and over-glazing techniques were used. Color parameters were tested by cs-5 spectrophotometer with CIE1976LAB color system before and after glazing. Statistical analysis was done and the color differences were calculated. The samples' surface structure was observed under the same background. RESULTS: No statistical significance existed in all of the color parameters between two groups before and after glazing. The values of color difference were all smaller than 1.5. The samples had the same surface structure after treated by different glazing techniques. CONCLUSIONS: Glazing techniques had no effect on final color of porcelain-fused-to-titanium restoration. No matter what kind of glazing techniques were used, the same surface structure and color would be obtained as long as the same polish way was used before glazing. PMID- 12903528 TI - [Intrathecal synthesis of the anti-Hu antibody in patients with paraneoplastic encephalomyelitis or sensory neuronopathy]. AB - OBJECTIVE: To probe into the pathogenic mechanisms of paraneoplastic encephalomyelitis (PEM) or sensory neuronopathy(PSN). METHODS: The serum and cerebral spinal fluid(CSF) anti-Hu antibodies and titers in 16 patients with PEM and 14 patients with PSN were detected with indirect immunohistochemistry (ABC) method. Intrathecal IgG synthesis and antibody specific activity (ASA) were calculated with Schuller's formulas according to the titers of the anti-Hu antibodies in the serum and CSF of the patients. Anti-Hu antibodies were identified by detecting the antigens of the nuclear extracts from the human cerebral cortex neurons with Western blot analysis. IgG in the serum and CSF and the albumin in the CSF were tested with electroimmunodiffusion method. RESULTS: The serum and CSF anti-Hu antibody titers in patients with PEM were significantly higher than those in patients with PSN (P < 0.001). A ratio of intrathecal ASA/serum ASA of anti-Hu antibody > 2 in 14 (88%) of the 16 patients with PEM and one (7%) patient with PSN indicated that the patients with PEM had a positive intrathecal anti-Hu antibody synthesis (P < 0.001). The Rankin scores are 4.5 +/- 1.7 in patients with PEM and 3.0 +/- 1.4 in patients with PSN (P < 0.01). Patients with PEM were more disabled than those with PSN. CONCLUSIONS: The detection of intrathecal ASA/serum ASA ratio for identifying the anti-Hu antibody intrathecal synthesis is valuable in judging the severity of disease and prognosis in patients with paraneoplastic syndromes of nervous system. PMID- 12903529 TI - [CD45-gating for flow cytometric immunophenotyping of leukemia]. AB - OBJECTIVE: To establish a reliable and rapid multiparameter flow cytometric approach for leukemia immunophenotyping. METHODS: Leukemia blast cells which exhibit a character of low CD45 expression and low side scatter density are easily recognized by "CD45-SSC gating", we measured 46 patients and 3 normal persons of immunophenotyping by this gating method as well as traditional forwardscatter-side scatter (FSC-SSC) gating method. RESULTS: 5 of 46 immunophenotypic results from "FSC-SSC gating" showed false diagnosis, which were corrected by "CD45-SSC gating". Further, immunophenotypic mapping of 46 different acute leukemia were obtained by "CD45-SSC gating" and three-color staining, The expression of related antigen could be readily observed. CONCLUSIONS: "CD45-SSC gating" is a reliable and easy method to subclassify leukemia. Three-color staining, which is able to analyze the expression among two related antigens, not only be helpful in leukemia diagnosis but also contribute to study the occurrence, development and prognosis of diseases. PMID- 12903530 TI - [Expression of tissue factor and vascular endothelial growth factor in colorectal carcinoma]. AB - Tissue factor (TF) and vascular endothelial growth factor (VEGF) play an important role in tumor progression and metastasis. We analyzed their expression in the carcinoma and normal mucosa of 53 colorectal cancer patients. VEGF levels were significantly higher in the tumor and correlated with TF expression. No correlation was found with tumor stage. TF may influence tumor growth and metastasis by modulating VEGF expression and neoangiogenesis. PMID- 12903532 TI - [Effect of preoperative biliary drainage on the immediate and long-term results after duodeno-cephalo-pancreatectomy for peri-ampullar neoplasia]. AB - Pancreaticoduodenectomy has a primary role in the treatment of patients with periampullary carcinoma. Several series have reported encouraging operative mortality and survival after resection. Controversies exist regarding the impact of preoperative biliary drainage on immediate and long-term results. PMID- 12903531 TI - [Neoadjuvant radiochemotherapy treatment in locally advanced rectal adenocarcinoma]. AB - A prospective phase II study was conducted to evacuate toxicity and results of preoperative radiochemotherapy in locoregionally advanced rectal cancer (LARC). A total of 33 patients entered the study and received 45 Gy to the pelvis plus a supplemental boost of 5.4-9 Gy concurrently with 5 FU c.i. at a dose of 225-275 mg/m2. Thirty patients were operated after 5-7 weeks (20 anterior resection and 10 abdominoperineal excision). In 14 patients (47%) a downstaging was observed, 5 patients experienced a complete clearance of the primary tumor. After a median of 14 months (range, 5-27), 23 patients, are alive and well. And 8 patients experienced a disease progression (4 local-regional and 4 distant). Our results provide further evidence of the utility and effectiveness of preoperative radiochemotherapy in LARC. PMID- 12903533 TI - [GIST of the jejuno-ileal tract]. AB - Jejunum-ileum stromal tumors give diagnostic and therapeutical problems, especially with regard to histopathological identification and biological behavior. These tumors are very rare in all case report. After gastric stromal tumors these ones are the most frequent of all alimentary tract. They are often incidentally found; because of their potential transformation, they have to be treated by respecting oncologic eradication standards and observing prognostic criteria based on a correct histopathological evaluation (neoplasm localization and dimension, its structure, presence of cytological atypisms, mitotic index, ploidia, surgical resection). Then there has to be a good follow-up program. PMID- 12903535 TI - [Radiofrequency wave treatment of liver tumors]. AB - The surgical therapy modifies the survival of the patients affections from tumor of the liver, primitive or secondary. The concomitance of cirrhosis, the dimensions, center and the distribution of the tumor reduces notably the number of the patients that they could be subjected to resection. The authors expose their experience with the employment of the radiofrequency thermoablation in the cases not tractable by means of surgery. PMID- 12903534 TI - [Perianal Bowen's disease: a case report and review of the literature]. AB - Perianal Bowen's disease is a uncommon, slow growing, intraepidermal squamous cell carcinoma (carcinoma in situ) of the anal region and may be a precursor to squamous carcinoma of the anus. It is associated with cervical and vulvar intraepithelial neoplasia and have human papillomavirus as a common cause. Both sexes and all races are affected, with the highest prevalence in patients aged 20 to 45 years. The symptoms of anal Bowen's disease are unspecific and the clinical findings are uncharacteristic and include pain, itching, bleeding and a disturbing lump. Biopsy and histopathologic examination is required for diagnosis and to distinguish other perianal dermatoses; thus an anogenital warts that fail to respond to conventional therapy, or change in appearance, warrant a biopsy and, where the technique is available, DNA typing to identify the viral pathogen. Infact the etiologic agent, the human papillomavirus (HPV), has been classified by DNA techniques into at least 42 types, of which 16 and 18 are considered to carry a high risk for cancer. The intraoperative findings is a lesion at the anocutaneous line: perianal or intra-anal tumor, erosion or ulceration as well as lichenoid lesion or hyperpigmentation. The disease has a proclivity for recurrence and there are many controversies concerning treatment that effectiveness remains uncertain and range from aggressive wide local excision with skin grafting when necessary to laser vaporization (argon or CO2), radiotherapy or a new immune response modifier (Imiquimod). We report a case of a 50-years-old woman with recurrence of Bowen's disease associated with vulvar HPV infection and review the literature. PMID- 12903538 TI - [172 consecutive total gastrectomies for gastric cancer with Roux-en-Y loop reconstruction and without fistulization of the esophago-jejunal anastomosis]. AB - In the treatment of gastric cancer total gastrectomy (TG) is one of the most important operation and the esophagojejunal anastomotic leakage is the most important early complication. In our series of 172 consecutive TGs (all controlled with Gastrografin-Rx in fifth postoperative day) we did not have any anastomotic fistula, independently from age, stage, type of limphadenectomy and visceral demolition. We believe that the correct technical performance is the most important factor in the esophagojejunal anastomosis. PMID- 12903537 TI - [The role of endoscopic surgery in respiratory tract neoplasia]. AB - Laser therapy--either by contact or non-contact method--can be considered the best palliative option for patients affected by airway obstruction due to malignant tumors considered inoperable and/or in emergency because of inadequate ventilation. The Nd-YAG laser is widely used in endoscopy. From 1988 to September 2002 at the Department of Surgery "P Valdoni" of the University of Rome "La Sapienza" 251 clinical cases affected by airway obstruction caused by primary or metastatic malignant tumors are treated; moreover 76 stents were used. The symptomatological improvement and beneficial changes in functional breathing tests, haemogasanalysis and X-ray findings were registered 48-72 hours after each treatment. The results showed an improvement in functional breathing values especially after treatment of the main airways such as trachea and main bronchi. The therapy offered an acceptable quality of life. A fairly good number of patients may also return to radio and/or chemotherapy (if previously interrupted) and sometimes to radical surgery. In terms of costs-effectiveness, the endoscopic treatment is widely competitive. PMID- 12903536 TI - [Indications for integrated surgical treatment of peritoneal carcinomatosis of colorectal origin: experience of the Italian Society of Locoregional Integrated Therapy in Oncology]. AB - A multicentric prospective study has been carried on 69 patients affected by peritoneal carcinomatosis from colorectal cancer. Patients have been treated by cytoreductive surgery and intraoperative hyperthermic chemoperfusion. CC 0-1 has been achieved in 82%. Major morbidity and mortality was respectively 21.7% and 2.9%. Three years overall survival was 26.7% for all series. Difference in survival evaluating CC 0-1 vs CC 2 patients and PCI < or = 10 vs > 10 was statistically significant. Evaluating only patients CC 0-1 and PCI < or = 10 overall survival rised up to 44.7% at 4 years. A smaller subgroup of patients with a disease-free interval to peritoneal carcinomatosis > or = 2-year showed a 50% disease-free survival at 5 years. In conclusion PCI < or = 10, complete or optimal cytoreduction feasibility and disease-free interval have to be considered for the patients selection to the integrate treatment. PMID- 12903539 TI - [Radiofrequency thermal ablation of hepatic nodules: selection criteria for percutaneous treatment]. AB - Radiofrequency thermal ablation (RFA) of liver tumor is done by percutaneous, laparoscopic and open surgical approach. Selection criteria for percutaneous or open surgical ablation of 54 hepatic lesions are here evaluated in 30 consecutive patients. Open surgical approach was performed in 9 cases only, 5 of them due to concomitant treatment of associated diseases. Number and size of the lesions did not interfere with surgical approach. Postoperative CT control showed no differences in terms of complete ablation of the tumor in between the two groups of patients. Percutaneous approach of RFA is gone to be in the future the modality of choice in these patients. PMID- 12903540 TI - [Intraperitoneal neoplastic dissemination of incidental gallbladder carcinoma after laparoscopic surgery]. AB - BACKGROUND: Several laboratory and clinical studies have raised the concern that laparoscopic procedures might worsen the prognosis of patient with malignant disease due to premature intraperitoneal cancer cell dissemination. The aim of the study was to examine the risk of the dissemination in patients with unsuspected gallbladder cancer diagnosed after laparoscopic cholecystectomy (LC). METHODS: A retrospective clinicopathologic study was performed on 5 patients with unsuspected gallbladder cancer without preoperatively diagnosis among 1280 patients that underwent LC in the period 1993-2002. The possible neoplastic dissemination was compared to that reported in 8 patients affected from unsuspected gallbladder cancer diagnosed after open cholecystectomy. RESULTS: Two patients with unsuspected gallbladder cancer (T1a-T2) presented a port-site recurrence after respectively six and four months. There was not statistically significant difference between types of cholecystectomy (LC or OC). Survival rate did not change according to whether the operation was carried out using LC or OC. CONCLUSION: After an accurate checking of the factors indicated to explain the early neoplastic dissemination frequently observed in the patients undergone LC and an analysis of their own experience the Authors conclude that LC does not worsen the prognosis of unsuspected gallbladder cancer. PMID- 12903541 TI - [Treatment of peritoneal mesothelioma using cytoreduction and intraperitoneal hyperthermic chemotherapy]. AB - Malignant mesothelioma of the peritoneum is a rare tumor for which the therapeutic approach has not yet been standardized. Cytoreductive surgery and hyperthermic antiblastic peritoneal perfusion (HAPP) may be effective in the treatment of this neoplasm. From 1995 to March 2003, we operated 24 patients with malignant peritoneal mesothelioma. Nineteen times we were able to perform radical surgery and HAPP: 6 CC-0, 10 CC-1 and 3 CC-2 (nodules smaller than 1 cm). HAPP was performed by the original "semi-closed" technique, using MMC+ CDDP in 3 patients, CDDP + doxorubicin in 15 patients, only doxorubicin in 1 patient). RESULTS: Operating mortality was 11% and postoperative morbidity was 26%. 4 patients are DOD (40, 20, and 2 at 2 months); 5 patients are AWD (72, 34, 25, and 2 at 15 months); 8 patients are NED (81, 47, 2 at 20 months, 16, 13, 12 months, 1 patient recently operated). The median survival is 40 months. The high rate of morbidity and mortality is due to the presence of postoperative paraneoplastic syndromes, with alterations of coagulation and onset of ARDS. Therefore the good survival results make the methodic one of the best options in the treatment of the disease. PMID- 12903542 TI - [Treatment of pseudomyxoma peritonei using cytoreduction and intraperitoneal hyperthermic chemotherapy]. AB - Pseudomixoma peritonei (PMP) is a rare neoplasia with a low grade of clinical malignity. Generally, the main treatment of this tumor is the surgical debulking. Best results are obtained combining surgery and hyperthermic antiblastic peritoneal perfusion (HAPP) with CDDP and MMC. From April '97 to March 2003, we operated on 132 patients, 8 times with a palliative intent. In 27 times we achieved a complete cytoreduction (17 CC0 and 10 CC1) followed by HAPP. As regards results, no post-operative mortality was reported and 19% of major morbidity was observed. 26 patients are NED at maximum follow-up of 6 years and 1 patient had recurrence 6 months after primary resection. We believe that cytoreduction and HAPP is the golden standard of PMP therapy when it is possible to achieve a complete cytoreduction. Most of the times, the disease is not radically treated and therefore, after diagnosis, patients should be only referred to specialized centers. PMID- 12903543 TI - [Reconstruction after gastric surgery: utilization of surgical staples and their optimization]. PMID- 12903544 TI - [Early evaluation using PET-FDG of the efficiency of neoadjuvant radiochemotherapy treatment in locally advanced neoplasia of the lower rectum]. AB - BACKGROUND AND PURPOSE: Preoperative chemoradiation allows downstaging of locally advanced rectal cancer and in selected patients also a sufficient downsizing to ensure sphincter preservation. Selection of patients warranting a preoperative approach is improved by magnetic resonance imaging (MRI) which is able to define the involvement of mesorectal circumferential margin. Similarly it would be crucial to define the response to chemoradiation during the treatment but traditional morphologic imaging techniques may fail in differentiating neoplastic tissue from scarring. PET-FDG has been successfully used in the detection of metastatic colorectal cancer allowing imaging of deposits as small as 0.5 cm and may have a role in evaluating early response to chemoradiation. METHODS: In the present study, in patients with T3-T4 rectal cancer undergoing preoperative chemoradiation PET-FDG and flow cytometry analysis on endoscopic biopsy specimen have been performed before, during and after preoperative chemoradiation. RESULTS: Chemoradiation treatment has been successful in terms of downsizing and downstaging of the tumor. PET-FDG was able to demonstrate local response at only ten-fifteen days after the beginning of neoadjuvant therapy, also identifying non responding patients. CONCLUSIONS: FDG-PET may have a role in defining the response to chemoradiation and modulate the treatments strategy in patients with advanced rectal cancer. PMID- 12903545 TI - [Cytoreductive surgery (peritonectomy) and intraperitoneal hyperthermic chemotherapy: an innovative and effective approach to the treatment of pseudomyxoma peritonei]. AB - Pseudomyxoma peritonei is a rare disease characterized by a complete redistribution of mucin into the peritoneal cavity. Thirty three PMP patients referred to National Cancer Institute of Milan were submitted to 34 consecutive procedures of peritonectomy and intra peritoneal hyperthermic perfusion (IPHP). Five yrs overall survival has been 96% while 3 yrs PFS 68%. Morbility, mortality and toxicity were 18, 3 and 9% respectively. Peritonectomy associated with IPHP is the most indicated approach to cure patients with DPAM and IFG, subgroup of pseudomyxoma peritonei. PMID- 12903546 TI - [Peritoneal mesothelioma: results of a complicated and aggressive procedure incorporating peritonectomy and intraperitoneal hyperthermic chemotherapy, and prospects derived from bench-to-bedside research]. AB - Peritoneal mesothelioma (PM) is a rare primary peritoneal tumor conditioning a very poor quality of life and prognosis. Twenty eight patients (11 men and 17 women) with PM were referred to National Cancer Institute of Milan and submitted to 29 consecutive procedures of peritonectomy and intra peritoneal hyperthermic perfusion (IPHP). Five years overall survival, disease free survival and progression free survival have been 70, 63 and 51%, respectively. Morbidity and toxicity were 14 and 21% respectively. No mortality has been observed peritonectomy associated with IPHP is an effective approach to cure patients PM. PMID- 12903548 TI - [Effect of pain-related neurotransmitters on cellular growth in cancer of the pancreas]. PMID- 12903549 TI - [Surgical tactics in abdominal wall neoplasia]. AB - Abdominal wall neoplasms represent more or less 1% of human neoplasms in the adult. The authors reports their own experience based on 9 cases during over 20 years. The prognosis of these tumors is almost influenced from a lot of factors such as: histology, localization, staging, grading, sex, surgical margins, number of local recurrences. Abdominal wall neoplasm are less aggressive for compartmentalization of muscle layer and with a better prognosis because of their localization, and surgical opportunities of extensive resection (not less of 2 cm from tumor's macroscopic limits) allowed by modern prosthetic reconstruction techniques. Polipropilene seems to be the ideal material for such kind of reconstruction even if also mersilene, PTFEe and others were employed. PTFEe and Dual-meshes could be useful in those malignant tumors in which peritoneum resection is necessary. PMID- 12903547 TI - [Expression of connective tissue growth factor (CTGF) in pancreatic cancer]. PMID- 12903550 TI - [The modified "hanging maneuver" during orthotopic liver transplantation using a technique for conserving the inferior vena cava]. AB - We describe a modification of Belghiti's "liver hanging maneuver" applied to the last phase of hepatectomy during OLT with IVC preservation. The proposed maneuver provides a better exposition of the suprahepatic veins allowing an ortogonal clamping of the suprahepatic confluence and avoiding caval clamping. It allows, moreover, an increase of venous surface available for the anastomosis that results wider and easier to perform. This provides a large outflow anastomotic cloaca and prevents outflow problems of the graft. PMID- 12903552 TI - [D2 pancreas-preserving lymphadenectomy in tumors of the upper third of the stomach]. AB - The technique of the D2 pancreas preserving lymphoadenectomy in gastric cancer surgery is described. The technique allows the complete clearing of the node stations n. 11,10 without pancreatic tail resection, which avoids major complications. PMID- 12903551 TI - [Stage IV gastric cancer and FOLFOX 6. Case report]. PMID- 12903553 TI - [Staging videolaparoscopy of peritoneal carcinomatosis]. AB - The peritoneal surface malignancy is an high risk condition which implies high postoperative mortality also for simple explorative laparotomy. A technique of staging laparoscopy was assessed and is here described: the technique reveals to be safe, feasible and reliable. The accuracy of the procedure depends our the surgeons experience and familiarity with the concepts of PCI and cytoreduction. PMID- 12903554 TI - [Multimodal treatment of liver metastases: our experience]. AB - Our recent experience in multimodal treatment of liver metastases. Surgical treatment of liver metastases is increasing in these years regarding more and more patients, as well as more and more surgeons. In the same time the introduction of therapies allows today the integration of surgical treatment and also satisfactory palliation. Authors report their experience in these last 4 years and suggest interdisciplinary approach to get good results even in non dedicated centres. PMID- 12903555 TI - [Traditional esophagectomy and esophago-gastrectomy vs. laparoscopic surgery. Evaluation and results]. AB - The therapeutical choice in patients affected with esophagus disease depends on patients' conditions, as well as on the benign or malignant nature of the lesion. Actually, the surgical option is strongly conditioned by the state of health, particularly, the surgical therapy of resection, that is a mayor kind of surgery, has to value the heavy engagement to which the patient is submitted, meant as a clinical stress, caused by the clinical situation in which the patient can frequently feel: malnutrition, immunological and hematological conditions. It is in this field that the mininvasive surgery can be considered the most important surgical choice. The target of the treatment of the traditional and mininvasive surgery, to cure the intestinal transit, by means of other internal organs, is the same: that is curative or palliative. In this study we analyzed and compared 8 patients, from July 1999 to March 2003; all of them undergone a therapy of resection in order to cure esophageal lesions (we applicated the same technique of esophagectomy according to Orringer): 5 of them underwent a mininvasive surgery, while the other 3 patients underwent a traditional surgery. As a result we saw the same mortality in both groups, with a minor morbidity in the second one (mininvasive surgery); this fact is very important because we can suppose an increase in the number of patients threatened with a curative or a palliative surgery, thanks to a minor load of the mininvasive surgery compared to the same surgical operation performed with the traditional methodology. PMID- 12903556 TI - [Therapeutic strategy in carcinoma of the exocrine pancreas]. AB - A series of 311 consecutive patients undergone surgery for pancreatic carcinoma in the Department of General Surgery from July 1979 to March 2003. We performed 41 standard pancreaticoduodenal resections (13%: 30 DCP, 2 total pancreatectomies, 9 splenopancreatectomies), 235 by-passes (75.5%: 114 Roux-en-Y hepaticojejunostomies, 99 hepaticojejunostomies with GEA, 22 GEA), 35 explorations and biopsy (11.2%: 28 LE and 7 VLS). Mortality rate was: 2.4% in Resection, 3.7% in BB(+)-GEA, 18.8% in GEA, 0% in LE-VLS. Morbidity rate was: 43.9% in resection (pancreatic fistula 21.9, haemorrhage 12.1, pneumonia 4.8, infection and delayed gastric emptying 2.4), 10.3% in BB(+)-GEA, 27% in GEA, 5.7% in LE. Actual survival rate was at 3 and 5 years after resection 9.7% and 4.8% respectively with median 18 months; mean survival was after by-pass 11 months (min 3, max 38) with median 9 months. PMID- 12903557 TI - [Metal endoprosthesis in the treatment of acute neoplastic occlusion of the colon. Our experience]. AB - BACKGROUND: Acute large-bowel obstruction represents a surgical challenge whose management is controversial, usually requiring emergent surgery in adverse circumstances with increased morbidity and mortality compared with elective situations. METHODS: We report our experience with self-expanding metal stents in the initial management of acute neoplastic colorectal obstruction. From December 2000 to February 2003, insertion of self-expanding metal stents under endoscopic and fluoroscopic guidance was attempted in 22 patients, in 9 as primary palliative measure (group A) whereas in 13 as a bridge to surgery (group B). RESULTS: Stenting was technically successful in 20 patients. There were 2 perforations, and one patient died as a consequence. Nine of the 11 patients in the group B underwent elective colonic resection while in two patients with advanced malignancy the stent was considered as a definitive palliative treatment. In all 10 patients with ultimate prosthesis (8 group A, 2 group B) two dislocations and one recurrent obstruction were noted in the mean follow-up of 12 months. CONCLUSION: Self expanding metal stents represent a good option in the treatment of large bowel obstruction, providing time for a complete preoperative evaluation and mechanical bowel preparation as well as for restoring patient's general conditions thus avoiding emergency surgery with considerably lower morbidity and mortality rates. In patients with advanced cancer, they provide an alternative to surgery with satisfactory results. PMID- 12903559 TI - [Preoperative CEA: prognostic significance in colorectal carcinoma]. AB - The prognostic meaning of preoperative CEA level and its relation to the other risk factors are still under debate. In 512 patients who underwent surgical treatment for colorectal cancer, CEA preoperative plasma level had been evaluated. The prognostic value of CEA was compared with other prognostic factors and the characteristics of the tumor. There was no significant ratio between CEA overexpression and stage, diameter, grading, ploidy, site and shape of the cancers. As regard as the long-term results are concerned, the patients with normal preoperative CEA levels had a better prognosis. In the Dukes B and C tumors, the level of CEA over the cut off point lets identify a group of patients with high risk whom more aggressive adjuvant therapies and follow up could be addressed to. This study suggests that CEA preoperative is an independent prognostic factor and may be useful in the therapeutic planning. PMID- 12903558 TI - [Role of intensive follow-up in colorectal cancer surgery patients]. AB - Authors analyze the role of follow-up after curative resection for colo-rectal cancer. In 129 patients (29.6%) a recurrence was observed, whilst in 6 (1.4%) a metachronous carcinoma was detected by endoscopy. Liver involvement was observed in 59 patients, lung metastasis in 18 and both liver and lung metastasis in 4. In 23 patients local recurrence (LR) was observed and in 25 both LR and liver metastasis were detected. 31 patients (24%) were submitted to surgery and 21 (16.2%) with curative purpose. As far as LR is concerned, 5 patients were operated (10%) with curative purpose and a 5 years survival of 40% was observed. Five years survival in 9 patients submitted to surgery with curative purpose (8.4%) for liver metastasis was 57.1%, whilst in 7 patients operated for lung metastasis (31.8%) it was 76%. All the 6 patients operated for metachronous carcinoma are still alive with a 2-5 years follow-up. PMID- 12903560 TI - [Functional complications of the lower urinary tract after curative exeresis for cancer of the rectum]. AB - The aim of the study is to assess the incidence and the pathogenesis of urinary dysfunction in the surgical treatment of rectal cancer. A series of 219 patients, who underwent surgery for rectal cancer with normal urinary preoperative function, was submitted to a prospective follow-up for the urinary function. Symptomatic patients in the follow-up were studied by ultrasonography, cystomanometry and pressure-flow electromyography. In the immediate follow-up only 17 patients with urinary dysfunction were observed (14 at stage 2, 2 at stage 3 an1 at stage 4 according Saito classification). Six months later only 8 patients had urinary dysfunction. In 1 of these cases neurogenic bladder requiring catheterization was observed no significative correlation with staging, radiotherapy, size of tumor, surgical technique was observed. Worst functional results were observed in patients who underwent abdomino-perineal resections. PMID- 12903561 TI - [Gene expression of the MAGE-1, -3, and -6 antigens in hepatic carcinoma]. AB - MAGE is a family of genes specifically associated to human melanoma, but also found in hepatocellular carcinoma (HCC). In this study we evaluated the expression of such genes in 41 HCC patients, their correlation with pathological and clinical aspects of cancer, and the impact on prognosis. MAGE are expressed in most of HCC samples (28/41), no correlation was found with type and stage but they may be used as potential target for IT. PMID- 12903562 TI - [Anatomical hepatic resection using radiofrequency thermoablation in the treatment of primary or secondary liver tumors]. AB - Radiofrequency thermoablation (RFA) in performing parenchymal liver section has been tested. Eleven patients with primary cancer or colorectal cancer metastasis underwent anatomical major liver resection after vascular control. Parenchymal section was performed by knife after coagulative necrosis. No deaths, clinical or technical complications or blood losses occurred. Such technique is feasible and effective. PMID- 12903563 TI - [Combination of neoadjuvant chemotherapy, cryotherapy and surgery in the treatment of unresectable colorectal liver metastases. Long-term results]. PMID- 12903564 TI - [Role of transrectal echography in the diagnosis and treatment of villous tumors of the rectum]. PMID- 12903565 TI - [Combined treatment of inoperable liver metastases from colorectal cancer]. AB - AIMS: Liver resection improves survival in selected patients with colorectal liver metastases. However, the majority of patients with colorectal liver metastases have an inoperable oncological disease. The aim is to investigate whether intra-arterial infusion of chemotherapy, improves response to treatment and may convert a selected group of patients with irresectable liver metastases into an operable state. MATERIALS AND METHODS: Thirty-sex patients (pts) with inoperable hepatic metastases from colorectal cancer were treated with intra arterial chemotherapy, by angiographic technique. All patients underwent a short 5-FU-based locoregional infusion and the 13 non pretreated patients also received systemic therapy. Evaluation of response was made by CT scan. RESULTS: Total cycles administered angiographically: 132 (range, 1-11). There were no complications associated with the angiographic procedure and no cases of > grade 2 toxicity. One heavily pretreated pt experienced moderate cholangitis and superficial gastric erosion. Thirty-one pts were assessable (20 pretreated and 11 not); there was 1 complete response (CR), 3 partial remissions (PR), 2 stabilizations (SD) among non-pretreated pts (6/11; CR + PR + SD = 55%) and 1 PR and 8 SD among pretreated pts (9/20; PR + SD = 45%). The remaining 16 pts progressed. Four pts became eligible for radical hepatic resection (1 refused surgery and 3 patients were operated on). There was no peri-operative deaths. Median survival of these 3 pts was 24, 28 and 39+ months. CONCLUSIONS: Our data, even if based on a relatively small case series, appear to confirm effective local disease control in this clinical setting. Regional chemotherapy used singly or in combination with systemic chemotherapy may convert a selected group of patients with irresectable liver metastases to an oncological disease that can benefit from surgical treatment. PMID- 12903566 TI - [Videolaparoscopic oncologic surgery of the right colon: our experience]. AB - The authors present their experience in videolaparoscopic surgery for right colon cancer after two years of videolaparoscopic surgery for approach to colorectal cancer. In the period between March 2002 and March 2003 they have practiced 11 right emicolectomy by videolaparoscopic technique for neoplasms: 7 were males and 4 were females with age of about 70 years old. These case included: 7 right colon's cancers, 2 cecal cancers and two hepatic flexure cancers. One postoperative complications has been observed (9%) for bowel's occlusion for early adhesion syndrome. On the base of their experience the authors perform usually videolaparoscopic surgery (M.I.S.) for the approach to right colon cancer. PMID- 12903568 TI - [Multivisceral resection of advanced gastric cancer]. PMID- 12903567 TI - [Videolaparoscopic oncologic surgery of the left and right colon: our experience]. AB - The authors present their experience in videolaparoscopic surgery for colorectal cancer. In the period between November 2000 and march 2003 they have treated 31 patients by videolaparoscopic surgery for colorectal cancer: 17 were males and 14 were females with age included between 41 and 93 y.o. These case included: 15 rectal cancers, 12 sigmoid cancer, 3 cases of lineal flexura's cancers and I case of transverse cancer. Postoperative complications has been observed in three cases (9.6%) in the first 14 operations practiced, then confirming the learning curve. The authors perform usually mini-invasive surgery (MIS) for the approach to colorectal cancer surgery. PMID- 12903569 TI - [Laparoscopic surgery for cancer of the rectum]. PMID- 12903570 TI - [New thrombophilia markers in digestive tract neoplasia]. AB - Haemostatic system compounds not routinely studied, have been evaluated to define the individual risk of VTE (venous thromboembolism) and to influence the prognosis using selective drugs. Significantly high values of fibrinogen, free TFPI, F1 + 2 fragments and TAT complexes on coagulation side and PAI-1 and TAFI on fibrinolysis side have been detected. Thrombin seems to have a role in the inhibition of TAFI dependent fibrinolysis not inhibited by heparin. PMID- 12903572 TI - [Left pyo-pneumothorax: a rare complication of colon carcinoma]. AB - A case of pio-pneumothorax complicating a splenic flexure colonic carcinoma is herein presented. The patient was a 58 years old male and was submitted 3 months earlier to a colo-colic bypass for a locally advanced tumor infiltrating stomach, spleen, tail of the pancreas and left emidiaphragm. Few days before the admittance in our ward, he experienced fever, anorexia, and severe dispnoea. Treatment was a water seal drainage of the chest evacuating nearly 8 Liters of purulent material where Escherichia coli was found. Death occurred 2 weeks after drainage. From the analysis of the literature thoracic empyema is an extremely rare complication of colonic carcinoma: 5 other cases have been reported so far. Pathogenesis in half of the cases was due to septicemia and in the others to infectious local spreading. PMID- 12903571 TI - [GIST: surgical treatment of 34 cases]. AB - In our Department of General Surgery 34 patients underwent surgery for GIST from 1983 to 2002. In stomach (21 cases) we performed: 7 local resections, 7 partial gastric resections, 5 total gastrectomies, 1 total gastrectomy + splenopancreatectomy, 1 diagnostic laparoscopy with biopsy. In small bowel (5 cases) we performed a typical intestinal resection, while in colon localization (4 cases) 2 anterior colorectal resections and 2 right partial colectomies. In duodenal and omental localization (2 cases each) we performed local resection. There was no operative mortality or morbidity. Actual survival is 67.6%. Prognostic prediction on the basis of histological findings is difficult, thus after surgery we suggest a long last follow-up. PMID- 12903573 TI - Gastric cancer in elderly and young patients: a Western experience. PMID- 12903574 TI - [Surgical treatment of gastrointestinal stromal tumors: personal cases]. AB - Surgical management of gastrointestinal stromal tumors: our experience. Gastrointestinal stromal tumors (GIST) are rare neoplasm even if they are the most common mesenchimal malignancies of the gastrointestinal tract. GIST have long been a source of confusion and controversy, in particular to define their pathological classification, preoperative diagnosis, management strategies, and prognosis. A radical resection of the tumor is often possible and always recommended. Recent studies have identified its principal pathogenic defect. An effective adjuvant therapy treatment (STI571) have been found for advanced and metastatic GIST. PMID- 12903576 TI - [Postoperative complications and local recurrence of mid-lower rectal carcinoma treated with neoadjuvant chemoradiotherapy and surgery. Preliminary results]. AB - In our study we evaluate postoperative complications and local recurrence in local advanced rectal cancer after preoperative chemoradiation plus surgery. We treated 24 patients, 15 males and 9 females; in all cases the cancer was at stage II or III. Down-staging was observed in 14 patients (58.3%), 2 patients (8.3%) had no residual disease. We performed 2 APR, 20 low anterior resections (10 of which laparoscopic) and 2 transanal local excisions. In postoperative period we observed 8 complications, 4 minor and 4 major ones, with an over-all morbidity of 33.3% and of 16.6% if considering major complications. At present no local recurrence was registered. PMID- 12903575 TI - [Adenocarcinoma of the cardia: surgical strategies compared]. AB - BACKGROUND: The incidence of adenocarcinoma of the cardia is increasing. The choice of surgical strategy for AEG type II of Siewert classification is the subjects of controversial discussion. The aim of our retrospective study is to analyse the surgical results in 12 years. MATERIAL AND METHODS: Since 1990 to 2002, 111 patients underwent resection for adenocarcinoma of the cardia at III Division of General Surgery, University of Turin. Twenty-five patients had AEG type I, 39 had type II and 47 type III. Transthoracic or transhiatal oesophagectomy with resection of the proximal stomach were performed in 55 cases and extended total gastrectomy with transthoracic or transhiatal oesophagectomy in 56. RESULTS: The morbidity and mortality rates are 17% and 5.4%. The 5 years survival rate is poor (35%) for all Siewert type. Survival is significantly associated with stage and the presence of lymph node metastasis, but not correlated with Siewert classification and surgical approach. Also in the AEG II the survival is not modified by the surgical approach. CONCLUSION: In patient with AEG I the therapy of choice is a radical transthoracic or transhiatal oesophagectomy with resection of the proximal stomach. For type III extended total gastrectomy with transthoracic or transhiatal oesophagectomy is the procedure of choice. The superiority of the thoracoabdominal approach is therefore evident in terms of oncologic radicality. Survival is similar in AEG type II patients for both operations. A oesophagectomy with proximal gastric resection should be adopted for these tumors as the standard procedure in the majority of cases. PMID- 12903577 TI - [Locoregional chemotherapy of locally advanced breast cancer in a pre-treated patient: case report]. AB - We report of the use of intra-arterial chemotherapy in one case of locally advanced breast cancer, that had been systemically pre-treated. Locoregional chemotherapy was delivered via percutaneous access. The catheter tip was placed into the subclavian artery and into origin of the internal mammary artery; it was removed after every cycle of treatment. The schedule of chemotherapy was: epirubicin 30 mg/m2, mitomycin 7 mg/m2 and 5 fluouracil 1000 mg. Three cycles were administered, and the treatment was well tolerated. The patient responded to intra-arterial chemotherapy, and she subsequently underwent complete surgical resection. Intra-arterial chemotherapy for breast cancer in an uncommon approach to the treatment of locally advanced disease. Nevertheless, in selected cases, it could be a more effective therapeutic option for patients with systemic chemotherapy-resistant disease. PMID- 12903578 TI - [Are small-size depressed-type colorectal neoplasias really the expression of a early stage of colorectal cancer. Apropos of a clinical case]. AB - A route of colorectal cancer development other than the adenoma-carcinoma sequence has recently become an issue due to the discovery of the depressed-type early colorectal cancers. Despite the protruding shapes of depressed-type early colorectal cancers, they probably have biological characteristics which differ from those of the usual polyp lesions. They show more aggressive behavior than the polypoid type and can arise de novo. Depressed-type lesions, in contrast to flat-type or protruded-type lesions, tend to invade the submucosa rapidly, so it is better treat them surgically from the outset. We report a case of a small depressed-type colorectal cancer involving the caecum of a 79-year-old male patient. PMID- 12903579 TI - [Acute liver toxicity of antiretroviral therapy (HAART) after liver transplantation in a patient with HIV-HCV coinfection and associated hepatocarcinoma (HCC)]. AB - OLT in HIV infected patients still remains a challenging option requiring a careful monitoring of patients for HCV reinfection, drug interactions and antiretroviral toxicity. Severe adverse events due to HAART have been already reported for post exposure prophylaxis in HIV infected patients. Here we report a case of liver graft toxicity related to HAART in a HIV-HCV co-infected patient (46 yrs-male) with associated a small HCC transplanted with a marginal liver graft. The patient had pre-OLT plasma HIV 1-RNA levels undetectable and CD4+ T cell count of > 200 cells/microL for 6 months. At day 2 a severe graft dysfunction was observed (AST 1570 U/L, ALT 2180 U/L, BIL tot 8.3 mg/dL, BIL Dir 6.6 mg/dL and PT 35%--INR 2.5). Doppler scan showed hepatic artery always patient. Later the postoperative in-hospital course was complicated by tense ascites and severe cholestasis. Serum bilirubin reached 42 mg/dL in day 12. Hypertransaminasemia ended at day 15 while cholestasis ended after 46 days. Tacrolimus was reintroduced at day 7. A liver biopsy 10 after OLT showed severe intrahepatic cholestasis, centrolobular necrosis and macrovesicular steatosis (30%). The patient was discharged 48 days after OLT with good liver function. After seven months HIV-RNA is still undetectable and HAART has not been restarted. We believe that the early complications we observed may be attributed to a sudden increase in plasma concentration of antiretroviral drugs secondary to drug redistribution from peripheral tissues and hepatic clearance deficiency after OLT. Although a pre-OLT withdrawal of HAART seems unjustified a delayed re introduction of HAART or the use of less hepatotoxic drugs may be advisable. PMID- 12903581 TI - [Total periareolar approach in breast-conserving surgery]. AB - Plastic and oncological breast surgery have to be considered as two aspects of the same treatment. The term "oncoplastic surgery" refers to the use of plastic surgery techniques in breast cancer surgery, in order to avoid and to correct the adverse aesthetic findings. The care of cosmetic sequelae of breast cancer surgery has reached an important therapeutic role for psychological consequences of disease and because of the higher patients expectations of a good aesthetic result. Considering the concept of oncoplastic surgery, since 1999 the authors began to use a periareolar approach in the breast conserving therapy (BCT), associated to axillary dissection performed through the same periareolar incision. This technique is not different from the traditional quadrantectomy in the extension of the glandular resection, while the skin may be preserved in according to the conventional protocols of BCT. Oncological and aesthetic results have proved to be safe and satisfactory. PMID- 12903580 TI - [Is it possible to improve survival in patients with Klatskin tumors?]. AB - BACKGROUND: This retrospective study was undertaken to evaluate if a high resectability rate could improve the long-term outcome of patients with proximal bile duct cancer. METHODS: Between 1985 and 2001, 50 patients (34 males and 16 females) with proximal bile duct cancer were treated. The tumor site were classified according to Bismuth-Corlette's classification: 9 lesions (18%) were Bismuth type I, 23 lesions (46%) type II, 12 lesions (24%) Type IIIa, 5 lesions (10%) type IIIb and only 1 resected tumor (2%) was type IV. Thirty-six patients (72%) were considered suitable for surgery, while 14 underwent non surgical palliative procedures. Twenty patients had bile duct resection only. Ten patients had Roux-enY cholangiojejunostomy with two or three divided segmental hepatic ducts; in ten, the cholangiojejunostomy was performed with 4 or 5 divided segmental hepatic ducts. Thirteen patients had bile duct resection plus hepatectomy. Despite the curative intention of the operation, only in 19 (52.7%) patients did the histopathological examination reveal tumor-free margins. RESULTS: There was no operative mortality. Overall 1-year, 3-year and 5-year survival of the entire surgical group was 61%, 22.5% and 9% respectively. In the 19 patients treated with curative intent the survival at 1,3, and 5 years was 66%, 30% and 15%, respectively, while in the palliative group it was 45%, 15% and 0%, respectively. CONCLUSION: Only margins free from tumor may guarantee an improvement in long-term outcome. Increasing resectability improves survival and could offer a chance of better 5-year survival. PMID- 12903582 TI - [Breast cancer screening in the elderly]. AB - The incidence and mortality of breast cancer increase with increasing age, and almost half of all breast cancer cases are diagnosed in women aged 65 years and older. Moreover many studies have indicated that the elderly are less screened, and have lesser and frequently inferior treatment. In contrast, clinical trials focusing on the elderly suggest that they do as well with surgery, radiation, and standard chemotherapy regimens as their younger counterparts. Our study suggest that in women aged 65 years, and older, breast cancer can be detected at an earlier stage by mammographic screening. PMID- 12903583 TI - [Bilateral breast cancer]. AB - Bilateral breast cancer has a cumulative incidence of 3-13% in patients with primary breast cancer, and the majority of these lesions are metachronous. The synchronous and metachronous bilateral breast cancer have at greater risk for distant metastasis than unilateral breast cancer. There was no difference in overall survival when comparing metachronous and synchronous bilateral patients to those with unilateral disease. PMID- 12903584 TI - [Intraoperative radiolocalization of intraductal breast carcinoma: sentinel lymph node evaluation]. AB - Sentinel lymph node biopsy and intraoperatory radiolocalization (RLI) are fast becoming the standard surgical treatment for invasive breast cancer. Actually, it seems, they have the same rule in "high risk" DCIS of the breast. PMID- 12903585 TI - [Intraoperative radiotherapy in the conservative treatment of initial-stage breast carcinoma]. PMID- 12903586 TI - [Chemotherapy in the treatment of breast carcinoma]. AB - Breast cancer (BC) is the most common malignancy and the second most common cause of cancer-related death in Western European and North American women. Neoadjuvant chemotherapy may be used in the management of both BC patients with locally advanced disease, and those with earlier stage and operable tumors. Data from recently phase III trials and worldwide consensus conference document the benefit of adjuvant chemotherapy in improving disease free survival and overall survival for patients diagnosed with invasive BC > 1 cm. When BC cells metastasize to distant organs, the disease is incurable, but chemotherapy may offer these patients a significant palliation. PMID- 12903587 TI - [Skin-sparing mastectomy and immediate reconstruction]. PMID- 12903589 TI - [Molecular therapy of breast carcinoma in the advanced phase]. AB - Conventional chemotherapy regimens for the treatment of breast cancer have limited efficacy and are associated with significant toxicity, highlighting the need for novel targeted therapies. Increased expression and activation of receptor tyrosine kinases frequently occurs in human breast carcinomas and, therefore, several clinical trials are currently evaluating therapies targeting these receptors. Therapeutic strategies include blockade of individual receptors with monoclonal antibodies (e.g., trastuzumab) and inhibition of tyrosine kinase function (e.g., gefitinib). Trastuzumab is the first agent that has been approved for patients with human epidermal growth factor receptor 2 (HER2)-overexpressing breast cancer. Other growth-factor targeted drugs are in clinical development such as STI-571, farnesyl-transferase inhibitors and antibodies directed at the insulin-like growth factor. PMID- 12903588 TI - [Diagnosis and treatment of non-palpable breast lesions: from the biopsy to the microhistology results. Our experience]. AB - The aim of modern senology lies in the diagnosis and treatment of non-palpable breast lesions (NPBLs). Through the diffusion of regional mammography screening the lesions being observed are continuously smaller, thus calling for more and more accurate methodology. Our experience in this area is based on the use of certain methods for retrieval and removal of NPBLs, such as Kopan's sec. philo guide, ultrasound and advanced breast biopsy instrumentation. In our opinion methods allowing total removal of lesions in order to obtain complete histopathological characterization and enabling adequate therapeutic programs are to be preferred. In reviewing case studies a noteworthy increase of initial carcinoma (DCIS or LCIS), from 19.5% to 57.1%, has been observed in the last three years due to the extensive use of the aforementioned methods. PMID- 12903590 TI - [Extemporaneous examination of the sentinel lymph node in breast cancer: is the glass half full or half empty?]. AB - Intra-operative examination of sentinel LN is controversial. Concordance with definitive exam of SLN in this series was 81%, though only 54% of positive cases were diagnosed. Micrometastases and ITC were usually lost intraoperatively, accounting for 14% of cases. Frozen section and touch prep of the SLN were approximately equivalent. The latter has the advantage of preserving tissue for step-analysis of SLN. The ultimate method of intraoperative analysis of SLN which can combine cost-effectiveness and accuracy needs to be determined. PMID- 12903591 TI - [Correlation between surgical intervention, adjuvant therapy, and long-term survival in patients operated on for breast neoplasia]. AB - INTRODUCTION: Breast cancer is the most frequent cancer among female sex, above all in rich countries where it occurs ten times more than the others. The most incidence of breast cancer is in female over 45 yrs and over 70 yrs with a plateau during menopause. So it is really very uncommon before 30 yrs. It is well known that breast cancer is related to several risk factors: these have always to be well evaluated before any treatment, especially in surgery. In the last years we can see an improvement of techniques to make diagnosis of breast cancer. Also surgery has improved in that way, so it can ensure better results to patients than before, also from the esthetical point of view. There are lot of discussions on which is the best surgical treatment to do, if lymphoadenectomy of the armpit is always required and if it has to be considered only as a prognostic element or also a therapeutical one. It is also important to plan radio, chemio and hormonal therapy, well chosen on the base of histological exams, and of the grading made either before either during surgery. Last studies moreover evaluate the specificity, the importance and the prognostic value of searching the so called "sentinel lymph node" who is always the first one to be interested in a metastatic process. STUDY AND DISCUSSION: Through a random and retrospective study among all our female patients (Department of Surgical Science in Rome University "La Sapienza") who had surgery for breast cancer in the last ten years (approving all the hystological exams) we have related the kind of surgical treatments to overall survival, to the therapies, and to any possible local recurrence of disease. We analyzed 270 female patients (age between 19 and 83). We considered all surgical treatments used in breast cancer: from the "simple" quadrantectomy up to radical mastectomy with the association of lymphoadenectomy of the armpit (both armpits in case of recurrence to disease). All these patients had then their own therapy (radio, chemo or hormonal one, it depended on each case) and follow up of the length of at least five years. The overall survival was of 82.2%. From these study we made important considerations about all the factors involved in breast cancer. The first step is to consider always all of these to have a well surgical approach, and to get the best compliance and performance from patients in order to have at distance the best results. PMID- 12903592 TI - [Can analysis of the molecular status of the p53 gene contribute to improving the therapeutic strategy for breast carcinoma?]. AB - The occurrence of mutations in the p53 tumor suppressor gene is a specific and recurring genetic event in solid tumors. P53 plays a pivotal role in multiple cellular processes such as cell growth control, DNA repair and programmed cell death. Genotoxic damage, also induced by chemotherapy or radiotherapy, induces p53 overexpression in order to control the rate of proliferating damaged cells, thus triggering the mismatch repair or apoptotic pathways. P53 inactivation determines a condition of genetic instability, justifying the subsequent susceptibility to acquire mutations of different other genes. P53 mutations are associated with worse prognosis and with chemo/radioresistance, due to the inability to trigger p53-dependent programmed cell death. Molecular diagnostic strategies show 32% p53 mutations in breast cancer. The analysis of the p53 gene performed by FAMA (Fluorescence Assisted Mismatch Analysis) in high-risk breast cancer patients with > or = 10 involved axillary nodes may help identify a subset of very high risk BC patients (vHR-BC) with poorer prognosis and a subset with better prognosis, potentially responsive to medical treatments. The accurate evaluation of the p53 status can predict prognosis and sensitivity to chemotherapy, thus representing the first step toward better definition of therapeutic strategies according to the molecular characterization of the individual patient. PMID- 12903593 TI - [Reliability of extemporaneous histological sentinel lymph node examination during breast surgery for neoplasia]. AB - Lymphatic mapping and sentinel lymph node (SLN) biopsy for breast cancer is rapidly becoming the standard of care. This is mainly due to the accuracy of the procedure, with a significant decrease in morbidity compared with the standard level III node dissection. We present our experience with SLN biopsy and a small series in which we performed an immediate histologic evaluation of the SLN: in case of positive SLN, a complete lymph node dissection was carried out in the same operative time, thus reducing the need of a second operation. In our experience, we had a 100% accordance between immediate and definitive results: we had neither false positive, which could lead to overtreat the patients with an unnecessary lymph node dissection, nor false negative. PMID- 12903594 TI - [Undifferentiated carcinoma of the thyroid]. AB - Anaplastic thyroid carcinoma is a rare and highly lethal cancer. Between 1969 and 2002, 9 cases with anaplastic carcinoma were reviewed. Surgical treatment with radiotherapy and chemotherapy were performed in all cases. All patients died of their disease within 5 years. The mean survival was 10 months. There is still controversy as to what constitutes adequate treatment for anaplastic thyroid carcinoma, and combined multimodal therapy seems to be the most common management strategy for this aggressive disease. PMID- 12903595 TI - [Parotid tumors: a diagnostic and therapeutic protocol in 81 surgical cases]. AB - AIM: In the last five years we cured 81 patients divided in two groups: the first was managed inordinately, the second according to a diagnostic and therapeutic protocol. METHODS: The first 23 patients were differently evaluated, before surgery, by means of traditional sialography, US, CT, CT sialography, MR, larger bore needles and FNAB. Neoplasm enucleations, atipic resections, superficial and total conservative parotidectomies were performed. The following 58 patients were evaluated and cured according to a diagnostic and therapeutic protocol. RESULTS: Diagnostic and therapeutic time were shortened, patients endurance improved and cost lowered. DISCUSSION: We think that many diagnostic procedures are useless and delay operation that, if possible, must be performed according to principles of radical extirpation. PMID- 12903596 TI - [Treatment for microcarcinoma of the thyroid. Clinical experience]. AB - According to WHO, small papillary cancer represents a papillary carcinoma < or = 1 cm in greatest dimension. The autoptic incidence is more elevated showing a low aggressivity. In fact lower mortality and local relapse have been reported. The AA report their results of 17 pt operated on for benign thyroid pathology in last three years. The diagnosis has been "incidental cancer" with 7 mm of mean diameter. Surgical treatment consisted of 9 total thyroidectomy, 3 near total thyroidectomy, 3 subtotal thyroidectomy, 2 lobo-isthmusectomy, with completion on the fifteenth day. Radioiodine therapy was administered in the case of uptake > 3.5%. There were no significant complications. Only in parathyroidectomy for secondary hyperparathyroidism, we have definitive hypoparathyroidism. Age, capsular infiltration, multifocality and lymphonodal dissemination represent main prognostic factors. In the treatment of SPC we recommend a total thyroidectomy or a near total in association with iodine-131 radioiodine therapy. PMID- 12903597 TI - [The right inferior laryngeal nerve with a non-recurrent course]. AB - Two cases of a right non-recurrent laryngeal nerve were encountered during the performance of 992 thyroid operations. In its abnormal non-recurrent course the nerve passes transversely from under the carotid sheat hand takes a position which is at right-angles to the normal recurrent laryngeal nerve. PMID- 12903598 TI - [Hormonal therapy in oncologic treatment: pathogenic hypotheses and interactions between thyroid and breast pathologies]. AB - INTRODUCTION: The aim to individuate the eventual correlation between the two pathologies has justified deeper studies to achieve new prospective approaches for both disease. BACKGROUND: We have selected 4 groups of patients who presented an association between the two pathologies: a) malignant breast pathology associated to a malignant thyroid pathology, b) patients with breast carcinoma who presented association with some thyroid alterations, c) patients with thyroid carcinoma who presented association with some breast alterations, d) patients who presented some associations between benign breast pathology and benign thyroid pathology. MATERIALS AND METHODS: We have excluded all patients with a clear physiological or surgical menopausal status, and we've so considered only patients with a regular menstrual cycle. We've so selected a group of 120 patients and we've performed in all these patients during the early follicular phase the following exams: breast echographic evaluation and thyroid echographic structure and volume determination and finally hormonal determinations we have so obtained two breast subgroups: 32 patients with hyperestrogenic integrative hormonal characteristics, 28 patients subjected to adjuvant hormonal therapy with hypoestregenic hormonal status and finally two thyroid subgroups, 22 patients showing clinical or subclinical hypothyroidism, 38 patients showing clinical or subclinical hyperthyroidism. We've compared these data to a random age-matched health control women group of 25 patients. RESULTS: The first group of patient showed a thyroid hormonal pattern of subclinical hypothyroidism or at least free T3 and free T4 mean value currently under and TSH and TPO Ab levels curve currently over the mean values of the control group. The second group showed the TSH suppressed with free T3 and free T4 curves currently over the mean value of the control group. The third group showed slight elevations in serum PRL levels curve. The fourth group showed increased estrogen levels-curve, often over the mean value of the control group. CONCLUSION: How much is it allowed to perform an hormonal therapy, specially for a benign pathology if we're not yet able to understand the deep and unknown interaction between breast and thyroid? PMID- 12903599 TI - [Metastatic thyroid tumors from adenocarcinoma of the colon: clinical and anatomo pathological contribution]. AB - We report herein the case of a 43 year-old man who was operated 1998 of left colectomy and hepatic resection for colic adenocarcinoma metastasized in the liver. He suffers of multinodular goiter from some years; was admitted to our hospital with a 2-month history of enlargement of goiter and appearance of a new nodule. Physical and imaging examinations disclosed a new nodules in the thyroid. Was operated of total thyroidectomia and lymphoadenectomia cervical in the December 2002. Hystologic examinations revealed adenocarcinoma which was consistent with a diagnosis of metastases from the primary colon adenocarcinoma to the thyroid. The rarity, diagnosis, and prognosis of thyroid metastasis from colon carcinoma are discussed. PMID- 12903600 TI - [Predictive factors for malignancy in Hurthle-cell thyroid neoplasia. Effect of surgical treatment]. AB - The aim of this study has been to evaluate factors predicting malignancy in patients with Hurthle cell neoplasms. Medical records from 36 patients who underwent thyroidectomy for Hurthle cell neoplasms between January 1998 and December 2002 were analyzed. Of the 36 patients, 19 had carcinomas and 17 had adenomas, resulting in a 52.7% prevalence of malignancy. Both fine-needle aspiration and intraoperative frozen section had low sensitivities in cancer detection (22.2% and 33.3% respectively). Hurthle cell carcinomas were significantly larger than adenomas (30.3 mm +/- 3.9 vs 17.6 mm +/- 2.3, P = 0.012), however 42% of carcinomas had a diameter between 10 and 20 mm. Size of Hurthle cell tumors is predictive of malignancy, but it is not the only factor to make surgical decision effective. Because of these uncertainties, authors believe that total thyroidectomy is the treatment of choice of all Hurthle cell neoplasms. PMID- 12903601 TI - [Liver metastases of choroid melanoma. Retrospective study of 63 cases]. PMID- 12903602 TI - [Tumors of the minor salivary glands located in the hard palate]. PMID- 12903603 TI - [The treatment of malignant pleural effusions: the experience of a multidisciplinary thoracic endoscopy group]. AB - More than half of neoplastic patients show in their clinical history the onset of pleural effusion. Malignant pleural effusion produces dyspnea, decreases respiratory function and quality of life in patients with advanced cancers. Optimal treatment is actually controversial. The aim of this study is to analyze the experience of malignant pleural effusion treatment of the Multidisciplinary Group of Thoracic Endoscopy. Patients are been subdivided in two group, depending on respiratory performance status and they are been submit to a Video-Assisted Thoracic Surgery (VATS) with talc pleurodesis and to positioning of a chronic indwelling pleural catheter. The treatment of malignant pleural effusion with the methods reported above allows, not only to achieve palliation of symptomatology, but also to achieve pleurodesis in patients with limited life-expectancy with good cost-beneficial ratio. PMID- 12903604 TI - [Hysteroscopy in the diagnosis of endometrial carcinoma]. AB - OBJECTIVE: To determine the diagnostic value of hysteroscopy in endometrial cancer. METHODS: 216 patients underwent hysteroscopy and endometrial biopsy. A comparison was made between hysteroscopic impression and histological examination. RESULTS: Sensitivity of hysteroscopy for diagnosing of endometrial cancer was 83.33%, specificity was 99.04%, positive predictive value was 71.42% and negative predictive value was 99.52%. CONCLUSION: Hysteroscopy was found to have a very important role in the diagnosis of endometrial cancer. PMID- 12903605 TI - [Tolerability of external electro-hyperthermia in the treatment of solid tumors]. PMID- 12903606 TI - [Anti-blastic hyperthermic perfusion in the treatment of melanoma of the extremities in the loco-regional diffusion phase]. AB - Primary limb melanoma may recur in terms of satellitosis, in transit metastases and/or regional node involvement. Hyperthermic antiblastic perfusion (HAP) permits the isolation of involved extremity from the systemic circulation and to deliver high doses of antineoplastic drugs. The association of cytostatic drugs to hyperthermia (> or = 41.5 degrees C) results in a synergistic effect with an increased therapeutic effectiveness. The overall 5 and 10-year survival rates in relation to the disease stages are st. II 75% and 67%; st. IIIA 59% and 42%; st. IIIAB 36% and 30% respectively. The results confirm that HAP is considered the treatment of choice of loco-regional spreading limb melanoma. Recently, the tumor necrosis factor (TNF) has been combined with Melphalan and hyperthermia. This trimodality association seems to be superior to Melphalan and hyperthermia alone only in patient with bulky tumors (i.e., multiple nodules), as a matter of fact the complete tumor response rates observed in these patients have been 67% and 20% respectively. The greater effectiveness of trimodality association has to be confirmed by multicentric randomized trials. PMID- 12903607 TI - [Utilization of a multidisciplinary protocol for postoperative pain treatment]. PMID- 12903608 TI - [A rare case of nerve-sheath sarcoma with rhabdomyoblastic differentiation (malignant triton tumor)]. AB - Malignant peripheral nerve sheath tumors (MPNST) are spindle-cell sarcomas that appear in a setting of neurofibroma or schwannoma or are associated with peripheral nerves or demonstrate nerve sheath differentiation. Malignant triton tumor (MTT) is a subtype of MPNST that also contain tissue with skeletal muscle differentiation (embryonal, plemorphic and botryoid rhabdomyosarcoma). The estimated incidence of MPNSTs in patients with NF1 is 2-5% compared with 0.0001% in the general population and approximately 69% of the reported cases of MTT are associated with von Recklinghausen disease. In July 2002 a 37-year old man was readmitted to the Department of Oncologic Surgery of the S. Camillo-Forlanini Hospital in Rome for both a right-sided retroperitoneal paravertebral not palpable mass, incidentally detected at a follow-up MRI, and a left-sided popliteal mass, discovered at clinical evaluation. Seventeen months before, when the patient underwent surgery at the same Department for both a left-sided paravertebral inferior mediastinal neurofibroma and a right-sided axillary neurofibroma, diagnosis of von Recklinghausen disease (NF1) was made, according to the criteria established by the NIH Consensus Development. Conference on Neurofibromatosis of 1987. A xifopubic laparotomy was performed: the tumor appeared to be localized, well-capsulated and strictly associated to the lumbar and sacral nervous radicles (L4, L5, S1) without evidence of invasion. The tumor was completely resected with sparing of the psoas muscle and the lumbar plexus through a subperineural dissection technique. No intra-operative pathologic examination was performed. Postoperative pathologic findings showed evidence for a trition tumor. The popliteal mass was resected too and resulted to be a neurofibroma just like the tumors resected 17 months before when diagnosis of von Recklinghausen disease was made. The patient was disease free 6 months after initial surgery. Sarcoma arising in anatomic site other than extremity and superficial trunk are often more difficult to control because of anatomic constraints, delayed disease presentation, proximity to neurovascular and osseous structures and toxicity for normal adjacent tissues that limits the use of adequate radiation doses. Indeed, the anatomic site is an important prognostic factor in STS and the prognosis for retroperitoneal tumors is considerably worse than for extremity tumors. Reported local recurrence rates for retroperitoneal sarcomas range from 40% to 80% and, in marked contrast to extremity STS, most of patients can and do die from local recurrence in the absence of metastasis. In contrast to the benefit most patients with high grade soft tissue sarcomas of the extremities receive from adjuvant radiation and chemotherapy, these modalities have been of little value for retroperitoneal tumors. To overcome the problem of dose limitation, intraoperative electron beam radiotherapy (IORT) in combination with ERBT has been proposed. IORT plus ERBT was found to improve local control of disease in recent clinical trials. Current chemotherapy for retroperitoneal sarcomas is ineffective. Local adjuvant therapy such as intraperitoneal chemotherapy or experimental immunotherapy seems to be attractive in theory, but needs further investigations through prospective randomized multicentric trials. In conclusion, to date aggressive surgical management remains the most effective modality for selected primary and recurrent retroperitoneal soft tissue sarcomas including MPNSTs and the subtype MTT. Patients with incomplete resection and other risk factors such as younger age and high grade tumors may be suitable candidates for investigational adjuvant therapy. PMID- 12903609 TI - [Hysteroscopy in suspected endometrial carcinoma: a comparison among cavity distention media]. AB - OBJECTIVE: To evaluate satisfaction rate, efficacy and complication rate of carbon dioxide (CO2) vs normal saline as uterine distension medium for outpatient diagnostic hysteroscopy in patients with suspicious endometrial cancer. METHODS: Thirty-eight undergoing outpatient hysteroscopy performed with carbon dioxide (group A) or normal saline (group B). Quality of vision of uterine cavity, procedure time, complications, patient discomfort, satisfaction rate have been evaluated. RESULTS: In group B a significant reduction in abdominal pain, shoulder tip pain and a lower incidence of vasovagal reactions were observed in comparison to group A. In normal saline group we found a higher satisfaction rate and a lower operative time in comparison to the CO2 group. CONCLUSIONS: Uterine distension with normal saline seems to have less adverse effects and is better accepted by patients. PMID- 12903610 TI - [Evaluation of endometrial carcinoma using hysteroscopy and transvaginal echography]. AB - OBJECTIVE: To compare transvaginal ultrasound and hysteroscopy in the diagnosis of endometrial cancer in women with postmenopausal bleeding. METHODS: Sixty postmenopausal women with abnormal uterin bleeding underwent to transvaginal ultrasound, hysteroscopy and endometrial biopsy. RESULTS: USG and hysteroscopy demonstrate respectively a sensitivity of 96% and 90%, specificity of 88% and 94%, positive predictive value of 86% and 92% and negative predictive value of 92% and 96% and accuracy of 89% and 94%. CONCLUSIONS: The accuracy of hysteroscopy is significantly higher than transvaginal sonography for the detection of endometrial cancer. PMID- 12903611 TI - [Laparoscopic adrenalectomy in the treatment of malignant adrenal lesions]. AB - The role of laparoscopic approach for the treatment of malignant adrenal diseases is still controversial. The aim of this study was to verify the results of laparoscopic adrenalectomy in the management of adrenal malignancies. The medical records of all the patients who underwent laparoscopic procedures for adrenal diseases and in whom malignancy was demonstrated at final histology were reviewed. Nine patients were included (3 malignant pheochromocytomas, 4 adrenocortical carcinomas and 2 adrenal metastases). At a mean follow-up of 17.0 +/- 12.8 months (range, 2-36), all but two patients were alive and disease free. One patient died for unrelated causes. No patient developed local or port site recurrence. The results of this study demonstrate that laparoscopic adrenalectomy can be safe and effective also in case of adrenal malignancies. Conversion to open surgery in mandatory in case of local invasion and when the dissection cannot be as accurate as in conventional operations. A preliminary laparoscopic exploration can be planned in case of suspected malignant lesions to confirm the diagnosis and to evaluate their operability. PMID- 12903613 TI - [The stapler in total laryngectomy with closed technique]. AB - AIMS AND BACKGROUNDS: The use of the stapler in total laryngectomy with closed technique is described by the authors. MATERIALS AND METHODS: From October 1999 to December 2002, 27 total laryngectomy were performed, 15 with a stapler and 5 cases had a primary tracheoesophageal puncture. RESULTS: Only one patient developed a pharyngocutaneous fistula, no other patients experienced post operative complications. All cases in this study had negative surgical margins. CONCLUSION: The correct indication for the use of the stapler is a tumor that needs a total laryngectomy, but completely endolaryngeal. The technique allows to remove the nso-gastric tube at the end of the first week after the operation, to avoid soiling the surgical field, to reduce the time in theatre. This technique seems to be safe as the traditional suture. PMID- 12903612 TI - [Treatment of the facial nerve and the neck in malignant parotid gland tumors]. AB - In the surgery of malignancies of the parotid gland the management of the facial nerve and of the neck is very important. In fact, many authors declared as the surgery of parotid gland can be considered as the surgery of facial nerve and others underlined the role of the neck management in a complete treatment of these neoplasms. The features of 47 cases of parotid gland neoplasms observed at the National Cancer Institut of Naples (Naples, Italy) are reported. In all cases a total parotidectomy was performed with neck dissection in case of neck metastases or electively in selected cases and resection of the facial nerve only in case of macroscopic tumor involvement. Radiotherapy was administrated in case of involvement of the facial nerve, and/or of the skin and/or of multiple neck metastases. The results of our experience emphasize as the neoplasms of the parotid gland can be treated sparing the facial nerve when it is clearly not involved without making worse prognosis; on the other side planing a neck dissection should be mandatory in case of high degree malignancies, and/or of tumors larger than T2, and/or of involvement of the facial nerve, and/or of the skin and/or cervical lymph nodes. PMID- 12903614 TI - [Endoscopic excisional biopsy with laser CO2 for the diagnosis and treatment of glottic carcinoma (T1-selected T2)]. AB - AIMS AND BACKGROUNDS: In T1-T2a glottic carcinoma before considering which type of treatment is the best option a biopsy is necessary. A peculiar aspect of this type of lesions is the fact that in the same specimen it is possible to find different grade of dysplasia even carcinoma in situ or microinvasive or invasive. An incisional biopsy could mislead the diagnosis. The aim of this study was to report the oncological results of the excisional biopsy for T1-T2a glottic cancer. MATERIALS AND METHODS: From November 1999 to December 2001, 18 patients (13 T1, 5 T2a) with carcinoma of the vocal cord underwent excisional biopsy of the lesion. In the case of negative margins no further treatment was done. The average follow-up was 2 years and 3 months. RESULTS: Local control was achieved in 100% for T1, 80% for T2. Radiotherapy was added in 2 cases. CONCLUSION: In our experience "excisional biopsy" for unadvanced glottic tumors is an ideal surgical technique for a one stage procedure of diagnosis and therapy. PMID- 12903615 TI - [Role of neoadjuvant chemotherapy in intermediate/high-grade soft-tissue sarcomas in the adult]. PMID- 12903616 TI - [Complete mediastinal dissection improves survival in patients with lung cancer]. PMID- 12903617 TI - [Clinical results in lung cancer patients undergoing a bronchoplastic surgical procedure]. PMID- 12903618 TI - [Role of surgery in patients with lung cancer and solitary metastases]. PMID- 12903619 TI - [Impact of sentinel lymph node biopsy on the treatment of melanoma]. AB - In the National Cancer Institute and S Pio X Hospital series we registered 981 patients with primary cutaneous melanoma and no evidence of clinically detectable regional node metastases underwent sentinel node (SN) dissection to microscopically define the tumor status of the regional lymph nodes. In 62.2% of cases, only one SN was detected; 26.4% of patients had two SNs and 11.4% had three or more SNs. A positive SNB was demonstrated in 18.1%. Analysis of survival indicated that the tumor status of the nodes was the most important prognostic factor. Breslow's thickness had a significant impact on survival in tumors of 4 mm or thicker, and ulceration dropped to a borderline significant P-value. To assess the tumor burden in positive SNB, all slides (148 SN pos) were reviewed. Twenty per cent of these patients had evidence of metastasis in other nodes. Of the remaining 80% with a single tumor-involved SN, 62% had a single metastatic deposit. Preliminary data from this study indicate that several subgroups may be identified among patients with 1 positive node, but adequate analysis of survival requires a larger number of patients and a multicentric study. PMID- 12903620 TI - [Role of crural inguinal lymphadenectomy and dynamic sentinel lymph node biopsy in lymph node staging in squamous-cell carcinoma of the penis. Our experience]. AB - PURPOSE: We determine the value of dynamic sentinel node biopsy for staging squamous cell carcinoma of the penis. MATERIALS AND METHODS: During a period of 2 years 17 patients with penile cancer was performed after lymphoscintigraphy pre and intraoperatively with gamma ray detection probe at biopsy of the sentinel node with the aid of perilesional administered patent blue dye. After 20 days regional lymph node dissection was performed in all patients. RESULTS: Sentinel node metastasis was found in 5 patients; in one case lymphoscintigraphy not visualized sentinel node. All negative node biopsy was confirmed with the regional lymph node dissection. No major complication did occur after sentinel node biopsy. CONCLUSIONS: Occult lymph node metastases can be detected by dynamic sentinel node biopsy including preoperative lymphoscintigraphy, vital dye and gamma ray detection probe with a sensitivity of 78%, predictive negative value of 100% and with low morbidity. PMID- 12903622 TI - [Lymph node biopsy in the study of lymphomas: from the incisional biopsy to videolaparoscopic lymphadenectomy]. AB - The role of the surgeon in the treatment of lymphoproliferative diseases is mainly addressed to histological diagnosis and staging. The aim of this study was to analyze the results of lymph node biopsies in patients with Hodgkin's disease (HD) and non-Hodgkin's lymphoma NHL). Between January 1992 and March 2003, 37 patients (17 males and 20 females, mean age 57 years, range 17-90) were submitted to a node biopsy to determine type of lymphoma and clinical staging: there were 8 HD and 29 NHL. In a single case laparoscopy was adopted to remove abdominal nodes; the procedure was uneventful and the patients discharged in the third postoperative day. The Authors stress the importance of the minimally invasive approach in the management of lymphoproliferative diseases. PMID- 12903621 TI - [Cervico-mediastinal carcinoma of the parathyroid: report of a case]. AB - Parathyroid carcinoma is a rare malignancy. We report the case of a 66 years old man referred to our department for a large substernal goiter suspected as a thyroid cancer associated with severe hyperparathyroidism. After normalization of serum levels of calcemia, total thyroidectomy and subtotal parathyroidectomy was performed. Histopathology revealed a parathyroid cancer of 450 g. PMID- 12903623 TI - [Percutaneous neurolysis of the celiac plexus under CT guidance in the invasive treatment of visceral pain caused by cancer]. AB - To verify the effectiveness and the incidence of complication in the transcutaneal celiac plexus block with CT-guided in the patient with intractable upper abdominal cancer, using alcoholic solutions to different concentrations (50% and 96%), previous insertion of the peridural catheter. From December 1997 to June 2002, studies were carried out on 24 patients with CT-guided percutaneous coeliac plexus neurolysis including 17 men and 7 women with inoperable abdominal malignancy and two with chronic pancreatitis. The patients were affected by very intense pain controllable only with high doses of analgesic narcotics. Before the procedure a catheter was installed in the peridurale space between L1-T12. To avoid general anesthesia, 40 mL of marcaine 0.5% was injected to relieve the back pain sometimes reported after the neurolysis, caused by the diffusion of alcohol in the coeliac plexus. This technique requires a posterior percutaneous procedural transaortic approach CT scan guided, to determine the correct position of the needle tips and the spread of neurolytic solution (40 mL of 96% + 3 mL of contrast medium) around the origin of the coeliac trunk's anatomical center of the plexus. The first 10 patients have received 40 mL of 50% ethyl alcohol + 3 mL of contrast medium. To evaluate the rate of the analgesia relief, a visual analogue pain score (VAS) was used before and 48 hours after the neurolysis. The percutaneous neurolysis of the celiac plexus is useful to relieve the pain in patients affected by cancer developing in upper abdomen. The CT-scan guide of the needle allows an omogeneous distribution of the contrast medium. The insertion of the peridural catheter made a complete analgesia and reduced the incidence of complications. Our method provided an excellent control of the pain in all patients. In our experience the pain relief was almost complete in patients treated with 96% ethyl alcohol solution (VAS from 8 before the treatment to 1, 48 hours after the treatment). The alcohol administered in elevated concentrations (96%), does not increase the incidence of complications. PMID- 12903625 TI - [Use of pentaglobin in the treatment of sepsis in the patient undergoing intraperitoneal hyperthermic chemotherapy]. AB - In patients subjected to cytoreduction and hyperthermic antiblastic peritoneal perfusion (HAPP), their immunocompromized conditions claim a very aggressive therapeutic approach in case of periotonitis and sepsis. Therefore, we use an adjuvant therapy of severe bacterial infections additional to antibiotic therapy by using Pentaglobin. It contains human plasma proteins, of which immunoglobulin at least 95%, with high rate of IgG, IgM and IgA, infused intravenously at 12 mL/h for 3 days continuously. From November 2000 to March 2003, we combined this approach for the treatment of peritonitis/sepsis in 11 patients. In 10 patients we obtained a complete control of the infective status in 8 days, while 1 patient died for MOFF. So, even with the exiguity of our casuistry, we obtained a better and more rapid clinical control of the patients respect to the previous period when the Pentaglobin was not used. PMID- 12903624 TI - [Total implantable venous systems in day surgery]. AB - Total implantable venous system (port-cath)in day surgery. AIMS: Most of patients underwent to a total implantable venous system (port-cath) can be treated in day surgery. METHODS: Since January 2000 to March 2003 we have implanted 31 port-a cath, preferring access in subclavia v. We've treated all patients in day surgery and we've performed a Rx-control with a medium follow up of two hour. RESULTS: We have not reported any early complication, only 3 late complications: a device malfunction, a "kinking" reservoir and an infection of the subcutaneous pocket. The last two complications have required the explantation of the device. In difficult catheterizations can be helpful to use a pediatric kit. The rules of an aseptic and antiseptic operative procedure and the utilization of a short term antibiotic prophylaxis can avoid infective complications. We have proposed, after the device implantation, cardioaspirina like preventive therapy of septic and no septic thrombosis. In the thrombotic occlusion of the catheter we propose the use of activase. In malfunction's prevention we propose a lateral placement (to the skin incision) of the subcutaneous pocket and a 45 degrees reservoir rotation. CONCLUSIONS: The total venous system (port-cath) implantation can be mostly performed in day surgery, improving the compliance of the patients with a low cost procedure. PMID- 12903626 TI - [Adjuvant adoptive immunotherapy in patients with stage III and resected stage IV melanoma: a pilot study]. AB - Adoptive immunotherapy trials with tumor infiltrating lymphocytes (TIL) and interleukin-2 (IL-2) were carried out in the treatment of advanced melanoma with a 34% of overall responses (OR). However, theoretically it should be of greater benefit as adjuvant therapy, especially in high-risk stages (stage III and resected stage IV). In a pilot study, 22 patients (aged 23-72 years) with stage III-IV melanoma who underwent radical metastasectomy were reinfused with TIL cultivated and expanded in vitro with IL-2 from surgically removed metastases. IL 2 (starting dose 12 x 10(6) IU/m2) was co-administered as a continuous infusion according to West's scheme. A total of 8/22 (36.3%) patients were disease-free (DF) at a median follow-up of 5 years. DF survival (DFS) and overall survival (OS) in the remaining 14 patients were 44% and 37% and 52% and 45% at 2 and 3 years, respectively. The CNS was the only site of disease recurrence in 57% of patients who relapsed. DF patients received a higher median dose of IL-2 than those who progressed (total dose 110 x 10(6) vs 86 x 10(6) IU/m2, respectively). The progressive reduction in IL-2 dosage allowed all patients to complete treatment without permanent grade 4 toxicity. The effects of tumor immunosuppression in lymphocytes inside the tumor (TCR z and e chains, p56lck, FAS and FAS-ligand) confirmed that the potential function of TIL, immunodepressed at the time of metastasectomy, was significantly restored after in vitro, culture with IL-2. Adjuvant adoptive immunotherapy with TIL and IL-2 seems to improve DFS and OS, in comparison with literature data. Further studies are required to determine its role in the adjuvant treatment of patients with high-risk melanoma. PMID- 12903627 TI - [Early de novo neoplasia after renal transplantation]. AB - INTRODUCTION: The chronic use of immunosuppressive therapy in transplant recipients to prevent acute rejection increases the long-term risk of cancer. The overall incidence of de novo malignancies (DNM) after kidney transplantation ranges from 6% to 11%. PATIENTS AND METHODS: Between January 2000 and December 2002, 135 renal and 3 combined kidney-pancreas transplantations were performed. RESULTS: Of 138 solid organ transplant recipients, a total of 16 (11.6%) cancers were diagnosed in 10 renal transplant recipients (7.2%). Six patients were male and three female, with a mean age of 47 years (range, 19-63, years). Tumor presented at a mean time of 14 months (range, 2-24, months) after transplantation. There were three patients with skin cancers, three with Kaposis's sarcoma, one with renal cell cancer, one with bladder carcinoma and one with breast cancer. CONCLUSIONS: Although the DNM occurs more frequently many years after a kidney transplantation, our experience demonstrated that they can occur early in the follow-up. Skin malignancies had the best prognosis, probably because of early detection and treatment. Kaposi's sarcoma benefits from reduction or cessation of immuno-suppression, but there is a higher risk of graft loss. Solid organ de novo malignancies are often more aggressive than in normal population, and the life expectancy of these recipients is very low. Careful long term screening protocols are needed for detection of such malignancies in an early stage. PMID- 12903628 TI - [A review of the study on Drynaria fortunei]. PMID- 12903629 TI - [Advances in the study on medicament portions of caihu]. PMID- 12903631 TI - Oncology nursing bibliography. PMID- 12903630 TI - Direct myocardial effects of fish oil on ischemia-reperfusion injury. Beyond lipid membrane composition? PMID- 12903632 TI - [Maintenance of function and organ impossible--what to do?]. PMID- 12903633 TI - [Hypopigmentation of the fundus oculi and pulmonary fibrosis]. PMID- 12903634 TI - [Recurrent pain with an irritation free "subconjunctival" foreign body]. PMID- 12903635 TI - [Remifentanil--reloaded]. PMID- 12903636 TI - Data indicate reduced incidence of Salmonella in raw meat, poultry. PMID- 12903637 TI - When's the right time to build a new building? PMID- 12903638 TI - Transferred patients hurt referral hospitals' quality ratings. PMID- 12903639 TI - Test provides easy tool to assess asthma control. PMID- 12903640 TI - Is keratinized tissue necessary around dental implants? PMID- 12903641 TI - Autopsies still detect important errors. PMID- 12903642 TI - Pharmacists can help reduce falls in the elderly through medication management. PMID- 12903643 TI - [The AIDS epidemic widens]. PMID- 12903644 TI - Mitochondria and their role in preconditioning's trigger phase. PMID- 12903647 TI - Liver Disease Research Branch. PMID- 12903645 TI - Strategic Science Symposium: Human Papillomavirus Testing-Are You Ready for a New Era in Cervical Cancer Screening? September 21-22, 2002, Rosemont, Illinois, USA. Proceedings. PMID- 12903648 TI - Abstraction in perceptual symbol systems. AB - After reviewing six senses of abstraction, this article focuses on abstractions that take the form of summary representations. Three central properties of these abstractions are established: ( i ) type-token interpretation; (ii) structured representation; and (iii) dynamic realization. Traditional theories of representation handle interpretation and structure well but are not sufficiently dynamical. Conversely, connectionist theories are exquisitely dynamic but have problems with structure. Perceptual symbol systems offer an approach that implements all three properties naturally. Within this framework, a loose collection of property and relation simulators develops to represent abstractions. Type-token interpretation results from binding a property simulator to a region of a perceived or simulated category member. Structured representation results from binding a configuration of property and relation simulators to multiple regions in an integrated manner. Dynamic realization results from applying different subsets of property and relation simulators to category members on different occasions. From this standpoint, there are no permanent or complete abstractions of a category in memory. Instead, abstraction is the skill to construct temporary online interpretations of a category's members. Although an infinite number of abstractions are possible, attractors develop for habitual approaches to interpretation. This approach provides new ways of thinking about abstraction phenomena in categorization, inference, background knowledge and learning. PMID- 12903649 TI - HDL Cholesterol: Metabolic Pathways and Drug Developments. 4th International Conference, March 2-4, 2003, Cambridge, MA, USA. PMID- 12903650 TI - Challenges of designing hepatic encephalopathy treatment trials. PMID- 12903651 TI - [Ictal signs--cerebral localization and propagation]. AB - In this manuscript, ictal signs are reported with regard to their relevance concerning cerebral localization. Limitations of so-called silent brain regions in this respect are also discussed. As a simple aid for clinical diagnosis, the nuclear shell structure model is demonstrated. PMID- 12903652 TI - Beyond learning fixed rules and social cues: abstraction in the social arena. AB - Abstraction is a central idea in many areas of physical comparative cognition such as categorization, numerical competence or problem solving. This idea, however, has rarely been applied to comparative social cognition. In this paper, I propose that the notion of abstraction can be applied to the social arena and become an important tool to investigate the social cognition and behaviour processes in animals. To make this point, I present recent evidence showing that chimpanzees know about what others can see and about what others intend. These data do not fit either low-level mechanisms based on stimulus-response associations or high-level explanations based on metarepresentational mechanisms such as false belief attribution. Instead, I argue that social abstraction, in particular the development of concepts such as seeing in others, is key to explaining the behaviour of our closest relative in a variety of situations. PMID- 12903653 TI - Abstraction and reformulation in artificial intelligence. AB - This paper contributes in two ways to the aims of this special issue on abstraction. The first is to show that there are compelling reasons motivating the use of abstraction in the purely computational realm of artificial intelligence. The second is to contribute to the overall discussion of the nature of abstraction by providing examples of the abstraction processes currently used in artificial intelligence. Although each type of abstraction is specific to a somewhat narrow context, it is hoped that collectively they illustrate the richness and variety of abstraction in its fullest sense. PMID- 12903654 TI - [Walter Poppelreuter (1886-1939). Remarks on the cover illustration]. PMID- 12903655 TI - The emergence of abstract ideas: evidence from networks and babies. AB - What is abstraction? In our view, abstraction is generalization. Specifically, we propose that abstract concepts emerge as the natural product of associative learning and generalization by similarity. We support this proposal by presenting evidence for two ideas: first, that children's knowledge about how categories are organized and how words refer to them can be explained as learned generalizations over specific experiences of words referring to categories; and second, that the path of concepts from concrete to more abstract can be observed throughout development and that even in their more abstract form, concepts retain some of their original sensory basis. We illustrate these two facts by examining, in two kinds of learners--networks and young children--the development of three abstract ideas: (i) the idea of word; (ii) the idea of object; and (iii) the idea of substance. PMID- 12903658 TI - Abstraction in mathematics. AB - Some current interpretations of abstraction in mathematical settings are examined from different perspectives, including history and learning. It is argued that abstraction is a complex concept and that it cannot be reduced to generalization or decontextualization only. In particular, the links between abstraction processes and the emergence of new objects are shown. The role that representations have in abstraction is discussed, taking into account both the historical and the educational perspectives. As languages play a major role in mathematics, some ideas from functional linguistics are applied to explain to what extent mathematical notations are to be considered abstract. Finally, abstraction is examined from the perspective of mathematics education, to show that the teaching ideas resulting from one-dimensional interpretations of abstraction have proved utterly unsuccessful. PMID- 12903657 TI - Visual categorization: accessing abstraction in non-human primates. AB - Evolution might have set the basic foundations for abstract mental representation long ago. Because of language, mental abilities would have reached different degrees of sophistication in mammals and in humans but would be, essentially, of the same nature. Thus, humans and animals might rely on the same basic mechanisms that could be masked in humans by the use of sophisticated strategies. In this paper, monkey and human abilities are compared in a variety of perceptual tasks including visual categorization to assess behavioural similarities and dissimilarities, and to determine the level of abstraction of monkeys' mental representations. The question of how these abstract representations might be encoded in the brain is then addressed. A comparative study of the neural processing underlying abstract cognitive operations in animals and humans might help to understand when abstraction emerged in the phylogenetic scale, and how it increased in complexity. PMID- 12903659 TI - Abstraction and art. AB - In a given social context, artistic creation comprises a set of processes, which relate to the activity of the artist and the activity of the spectator. Through these processes we see and understand that the world is vaster than it is said to be. Artistic processes are mediated experiences that open up the world. A successful work of art expresses a reality beyond actual reality: it suggests an unknown world using the means and the signs of the known world. Artistic practices incorporate the means of creation developed by science and technology and change forms as they change. Artists and the public follow different processes of abstraction at different levels, in the definition of the means of creation, of representation and of perception of a work of art. This paper examines how the processes of abstraction are used within the framework of the visual arts and abstract painting, which appeared during a period of growing importance for the processes of abstraction in science and technology, at the beginning of the twentieth century. The development of digital platforms and new man-machine interfaces allow multimedia creations. This is performed under the constraint of phases of multidisciplinary conceptualization using generic representation languages, which tend to abolish traditional frontiers between the arts: visual arts, drama, dance and music. PMID- 12903660 TI - Abstraction and context in concept representation. AB - This paper develops the notion of abstraction in the context of the psychology of concepts, and discusses its relation to context dependence in knowledge representation. Three general approaches to modelling conceptual knowledge from the domain of cognitive psychology are discussed, which serve to illustrate a theoretical dimension of increasing levels of abstraction. PMID- 12903661 TI - [Croatian physicians as members of the German Ophthalmology Society and relations of Croatian ophthalmologists to German ophthalmology]. PMID- 12903662 TI - Abstraction and natural language semantics. AB - According to the traditional view, a word prototypically denotes a class of objects sharing similar features, i.e. it results from an abstraction based on the detection of common properties in perceived entities. I explore here another idea: words result from abstraction of common premises in the rules governing our actions. I first argue that taking 'inference', instead of 'reference', as the basic issue in semantics does matter. I then discuss two phenomena that are, in my opinion, particularly difficult to analyse within the scope of traditional semantic theories: systematic polysemy and plurals. I conclude by a discussion of my approach, and by a summary of its main features. PMID- 12903664 TI - [The physician as manager. Organization and guidance in general practice]. PMID- 12903663 TI - [A nice business: an eyeglasses fantasy]. PMID- 12903665 TI - [Approval of satellite practices and secondary practice offices]. PMID- 12903666 TI - Abstraction from a sensori-motor perspective: can we get a quick hold on simple perception? AB - Two main types of dissociation can be considered in order to articulate action and abstraction. Vision for action and vision for perception are often described as dissociated systems at both anatomical and functional levels. Within this framework, abstraction should be specific to perceptual representation, whereas the action system would simply analyse the objective metric of space. By contrast, one may focus on dissociations within the action system. In this case, one will accept that action may involve abstract representations, at least during movement preparation. But a specific visuomotor level of processing can be described that appears to comply with the spatial properties of the relationship between the actor and the environment. This system would be specialized for fast movement guidance towards pre-defined goals. Such an automatic piloting system would thus be free of abstraction. PMID- 12903667 TI - Expression of EGF receptor in duodenum at weaning. PMID- 12903669 TI - The treatment of huge spontaneous portasystemic shunts in liver transplantation. PMID- 12903668 TI - The management of gastrointestinal infections caused by cytomegalovirus. PMID- 12903670 TI - Abstract and associatively based representations in human sequence learning. AB - We give an analysis of performance in an artificial neural network for which the claim had been made that it could learn abstract representations. Our argument is that this network is associative in nature, and cannot develop abstract representations. The network thus converges to a solution that is solely based on the statistical regularities of the training set. Inspired by human experiments that have shown that humans can engage in both associative (statistical) and abstract learning, we present a new, hybrid computational model that combines associative and more abstract, cognitive processes. To cross-validate the model we attempted to predict human behaviour in further experiments. One of these experiments reveals some evidence for the use of abstract representations, whereas the others provide evidence for associatively based performance. The predictions of the hybrid model stand in line with our empirical data. PMID- 12903671 TI - Abstraction in art with implications for perception. AB - The relationship between people and art is complex and intriguing. Of course, artworks are our creations; but in interesting and important ways, we are also created by our artworks. Our sense of the world is informed by the art we make and by the art we inherit and value, works that, in themselves, encode others' world views. This two-way effect is deeply rooted and art encodes and affects both a culture's ways of perceiving the world and its ways of remaking the world it perceives. The purpose of this paper is to indicate ways in which a study of abstraction in art can be used to discover insights into, to quote the call for papers for this issue, 'our perception of the world, acquired through experience' and 'the way concepts are formed and manipulated to achieve goals'. PMID- 12903672 TI - A grounded theory of abstraction in artificial intelligence. AB - In artificial intelligence, abstraction is commonly used to account for the use of various levels of details in a given representation language or the ability to change from one level to another while preserving useful properties. Abstraction has been mainly studied in problem solving, theorem proving, knowledge representation (in particular for spatial and temporal reasoning) and machine learning. In such contexts, abstraction is defined as a mapping between formalisms that reduces the computational complexity of the task at stake. By analysing the notion of abstraction from an information quantity point of view, we pinpoint the differences and the complementary role of reformulation and abstraction in any representation change. We contribute to extending the existing semantic theories of abstraction to be grounded on perception, where the notion of information quantity is easier to characterize formally. In the author's view, abstraction is best represented using abstraction operators, as they provide semantics for classifying different abstractions and support the automation of representation changes. The usefulness of a grounded theory of abstraction in the cartography domain is illustrated. Finally, the importance of explicitly representing abstraction for designing more autonomous and adaptive systems is discussed. PMID- 12903673 TI - Process for Assessment of Scientific Support for Claims on Food (PASSCLAIM): overall introduction. PMID- 12903674 TI - [The trail of AIDS in Peru]. AB - This historical study of the Aids epidemic in Peru between 1983 and 2000 focuses on legal responses and on medical studies and intervention, especially during the early 1980s, a decade characterized by serious economic crisis and terrorist violence. As elsewhere around the world, the appearance of Aids intensified interactions between scientists, public health specialists, journalists, politicians, and patients, with major influence on culture as well as on public health responses to the disease. In addition to retelling an important story, the study demonstrates the validity and usefulness of the historical perspective, which affords a better understanding of recent events in health and in society overall. PMID- 12903675 TI - Noise-induced autoimmune sensorineural hearing loss. AB - Typically, autoimmune sensorineural hearing loss has been described as a slowly progressive, asymmetric hearing loss that is responsive to medications traditionally used in the treatment of other autoimmune conditions. These medications include steroids and cytotoxic drugs. Inciting factors in autoimmune inner ear disease are rarely cited. We describe a case of episodic sudden hearing loss triggered consistently by environmental noise. The hearing loss was responsive to steroids at the time of each occurrence and was determined to be autoimmune. This case raises questions about the relationship between autoimmune inner ear disease and sensitivity to environmental noise that warrant further research. PMID- 12903677 TI - Pathogenesis and methods of treatment of Zenker's diverticulum. AB - In the literature, various theories concerning the cause of Zenker's diverticulum have been propounded. I believe that an anatomic predisposition plays a prominent role in the pathogenesis. This conclusion will be discussed in light of a study of the literature and characteristics of a series of 646 patients with Zenker's diverticulum. The current methods of treatment by an external surgical approach, as well as the (micro)endoscopic procedures, are described. PMID- 12903676 TI - Pathology of benign paroxysmal positional vertigo revisited. AB - The pathophysiology of benign paroxysmal positional vertigo (BPPV) is not completely understood. Although the concept of degenerated otoconia transforming the posterior canal (PC) crista into a gravity-sensitive sense organ has gained popular support, several temporal bone (TB) series have revealed similar deposits in normal TBs, suggesting they are a normal change in the aging labyrinth. Furthermore, some TBs from patients with BPPV do not contain particles in the posterior canal. Five TBs from patients with BPPV were studied quantitatively and qualitatively. A small PC cupular deposit was found in 1 TB, while none was seen in the other 4 TBs. The major pathological changes were 1) a 50% loss of ganglion cells in the superior vestibular division of all 5 TBs and 2) a 50% loss of neurons in the inferior division of 3 TBs, and a 30% loss in 2 TBs that contained abnormal saccular ganglion cells. These observations support a concept in the pathophysiology of BPPV that includes loss of the inhibitory effect of otolith organs on canal sense organs. PMID- 12903678 TI - Stomal recurrence following total laryngectomy: clinical and molecular analysis of a series. AB - The aim of the present study was to analyze the risk features and to discuss the preventive measures and treatment of stomal recurrence (SR) after total laryngectomy (TL), and to investigate the clonal relationship between the primary and recurrent lesions. We reviewed a series of 25 patients affected by SR after TL for laryngeal squamous cell carcinoma. A subset of 7 coupled primary and recurrent carcinomas were examined for microsatellite alterations that were used as a marker of genetic lineage. The incidence of SR was higher in patients with initial subglottic lesions (subglottic, transglottic, or glottic-subglottic tumors) than in those with other locations of laryngeal cancer (p = .013). In addition, the frequency of SR was significantly higher in patients who underwent preoperative tracheotomy, performed 48 hours or more before the TL, than in the group of patients who did not receive preoperative tracheotomy (23.3% versus 0.46%, p < .001). According to microsatellite analysis, of the 5 informative patients, 2 demonstrated discordant alterations in the recurrent tumor indicative of clonal heterogeneity, 2 demonstrated identical alterations, and 1 showed an additional alteration in the recurrent tumor. Our analysis supports the hypothesis that a subset of SRs after TL may be second primary tumors, genetically unrelated to the primary laryngeal lesion. PMID- 12903679 TI - Room tilt illusion as a manifestation of peripheral vestibular disorders. AB - Room tilt illusion is a subjective distortion of verticality with transient paradoxical rotation of the visual field, usually in the frontal plane. It might result from dysfunction of the vestibular pathways with subsequent contradictory vestibular, visual, and proprioceptive inputs and erroneous cortical integration. It has already been described in association with brain stem and cortical lesions, but reports of cases of peripheral origin are scarce. We report here 23 cases of room tilt illusion, all but 2 occurring in patients with either vestibular peripheral abnormalities or normal assessment findings. A review of the literature is presented, as well as a hypothesis addressing this phenomenon. PMID- 12903680 TI - Value-based approach to power-assisted adenoidectomy. AB - A recognized disadvantage of power-assisted adenoidectomy (PAA) is the increased patient charge associated with the disposable instrumentation. The elimination of pathological review of routine adenoid specimens may provide a means of offsetting the increased charge, as 1) unsuspected findings are rare, and 2) PAA specimens are too traumatized to provide the microscopic detail necessary to make an unsuspected diagnosis. The pathology reports of all adenoidectomy specimens removed over a 10-year period were reviewed and combined with previously published reviews in order to estimate the prevalence of unsuspected disease. The estimated prevalence of unsuspected diagnoses found by routine pathological review of adenoid specimens is 37 per 100,000 cases (95% confidence interval, 26 51). In a separate analysis, a pathologist blinded to the technique of adenoid removal assessed the tissue effects of curette adenoidectomy versus PAA. Significant tissue damage at the microscopic level was identified in 6 of 11 specimens removed with the power-assisted technique as compared to 0 of 11 specimens removed by curette (p = .03). Eliminating histopathologic review of routine adenoidectomy specimens can potentially offset the increased patient charge of PAA by 62%. Power-assisted adenoidectomy, however, should be avoided in nonroutine cases in which the potential for occult disease exists. PMID- 12903681 TI - Lateral upper arm free flap for primary reconstruction of pharyngeal defects in ablative oncological surgery. Report of six consecutive cases. AB - Free microvascular flaps are an established method for soft tissue reconstruction following ablative oncological surgery in the head and neck. Functional reconstructions of the hypopharynx and the pharyngoesophageal segment (PES) are of particular relevance, as they are highly demanding surgical procedures. So far, the radial forearm free flap (RFFF) and the free jejunal transfer have been the transplants predominantly used for this purpose. The lateral upper arm free flap (LUFF) presents an alternative method for the fasciocutaneous tissue transfer. We report on our experience with the LUFF in a 56-year-old male patient with a pT3pN0M0 squamous cell carcinoma of the hypopharynx. A pharyngocutaneous fistula developed 5 days after pharyngolaryngectomy with bilateral neck dissection. The fistula was localized between the pharyngeal constrictor muscle and the esophagus and was closed with an LUFF from the left arm. Excellent flap adaptation to the remaining pharyngeal mucosa was observed. Although the length of the vascular pedicle and the diameter of the vessels in the LUFF are smaller than those in the RFFF, neither pedicle length nor vessel diameter proved to be a problem. The LUFF can be recommended as a well-vascularized, relatively safe and reliable flap for reconstruction of tubular structures such as the hypopharynx and the PES after tumor ablation and as an alternative to the RFFF. The flexibility of the LUFF allows surgeons to reconstruct the anatomy of the lost soft tissues as adequately as possible. PMID- 12903682 TI - Effect of growth factors on hyaluronan production by canine vocal fold fibroblasts. AB - Hyaluronan (HYA) is considered to be a crucial factor in scarless wound healing and in maintaining tissue viscosity of the vocal fold lamina propria. In this study focusing on the effects of growth factors, we examined how HYA is produced and controlled in canine cultured vocal fold fibroblasts. Fibroblasts were taken from the lamina propria of the vocal folds of 8 dogs and cultured with and without growth factors. The production of HYA in the supernatant culture was quantitatively examined by enzyme-linked immunosorbent assay. Hepatocyte growth factor, epidermal growth factor, basic fibroblast growth factor, and transforming growth factor beta1 all stimulated HYA synthesis from vocal fold fibroblasts. These effects differed with the concentration of growth factors and the incubation period. We also examined how frequently the growth factors had to be administered in order to maintain appropriate levels of HYA. A single administration was sufficient to maintain appropriate HYA levels for at least 7 days. The present studies have demonstrated positive effects of growth factors in stimulating HYA production. Further in vivo study is needed to clarify the usefulness of these growth factors in the management of vocal fold scarring. PMID- 12903683 TI - Prevalence of nasal polyps in adults: the Skovde population-based study. AB - Patients with nasal polyps present repeatedly in otorhinolaryngology practices, but the prevalence of nasal polyps in the general population is not known. Our objective was to investigate the prevalence of nasal polyps in an adult Swedish population in relation to age, gender, asthma, and aspirin intolerance. A random sample of 1,900 inhabitants over the age of 20 years, stratified for age and gender, was drawn from the municipal population register in Skovde, Sweden, in December 2000. The subjects were called to clinical visits that included questions about rhinitis, asthma, and aspirin intolerance and examination by nasal endoscopy. In total, 1,387 volunteers (73% of the sample) were investigated. The sample size was adequate, with a good fit to the whole population. The prevalence of nasal polyps was 2.7% (95% confidence interval, 1.9 3.5), and polyps were more frequent in men (2.2 to 1), the elderly (5% at > or = 60 years of age), and asthmatics. Subjective symptoms of aspirin intolerance were not found to correlate with polyps. Nasal polyps were more common in adults than was stated by the a priori estimate. The Skovde population-based study is considered representative for the Swedish population. PMID- 12903684 TI - Videofluoroscopic and manometric evaluation of swallowing function in patients with multiple system atrophy. AB - We investigated swallowing function of 29 patients with multiple system atrophy (MSA) by videofluoroscopy and manometry. Abnormal findings in videofluoroscopy were generally consistent with those in Parkinson's disease. Although findings of videofluoroscopy were not correlated with a history of aspiration pneumonia, severity of disease was significantly correlated with a history of aspiration pneumonia. Oropharyngeal and hypopharyngeal swallowing pressures of the patients were decreased to 73.9 +/- 48.4 mm Hg and 85.3 +/- 42.9 mm Hg, respectively, both of which were significantly different from the pressures of the control group. Incomplete relaxation of the upper esophageal sphincter was seen in 23.1% of the MSA patients, all of whom had had MSA for more than 5 years. In conclusion, patients with MSA are at risk for aspiration pneumonia as disease severity increases, and the swallowing function of patients with more than 5 years' duration of MSA should be routinely followed up with both videofluoroscopy and manometry. PMID- 12903685 TI - Malignant peripheral nerve sheath tumor of the parotid gland. AB - Malignant peripheral nerve sheath tumor (MPNST) has been defined as any malignant tumor arising from or differentiating toward cells of the peripheral nerve sheath. We treated a case of MPNST arising from the right parotid gland that showed a highly aggressive course. We reviewed the English-language literature published since 1990 and found 142 cases of head and neck MPNST reported within the past 13 years. The results of the review suggested that MPNSTs may arise from any organs of the head and neck. Immunohistochemical analysis of various neural markers plays a significant role in the evaluation of the histologic diagnosis. Curative treatment based on radical resection of MPNSTs of head and neck origin is more difficult than treatment of MPNSTs of other origins. PMID- 12903686 TI - Extension of nasopharyngeal lymphoma to the middle and external ear. AB - We describe a patient with nasopharyngeal lymphoma who was admitted for chemotherapy. At admission, otoscopic examination revealed a large polyp occluding the left external ear canal. The polyp was removed, and histopathologic examination showed the same tumor that was found previously in the nasopharynx. Magnetic resonance imaging showed an infiltrative process that involved the left side of the nasopharynx and extended toward the left middle and external ear. To our knowledge, this is the first case report of a direct extension of nasopharyngeal lymphoma into the middle and external ear. PMID- 12903687 TI - Blue rubber bleb nevus syndrome: immunohistochemical study. AB - Blue rubber bleb nevus is a rare entity consisting of distinctive angiomas in the skin and gastrointestinal tract, leading to occult or profound gastrointestinal bleeding and chronic anemia. The efficacy has been documented of systemic treatment with corticoids, interferon, vincristine, and, more recently, subcutaneous octreotide in the presence of active lesion proliferation or disseminated intravascular coagulation. A case of blue rubber bleb nevus syndrome with oral hemangiomas in a 24-year-old woman is reported. The surgical specimens were subjected to immunohistochemical study, which showed all of the hemangiomas to be in an inactive phase. PMID- 12903688 TI - Acute edema of the tongue: a life-threatening condition. AB - Acute macroglossia is rare, but may cause upper airway obstruction requiring emergency intervention. The cause of the problem is often obscure. Edema of the tongue may be due to angioedema or to allergy. We present several cases of acute edema of the tongue, in 3 cases causing life-threatening airway obstruction. Among these, we present the first case of acute enlargement of the tongue due to the ingestion of artichoke. PMID- 12903689 TI - Fibrous dysplasia of the temporal bone. PMID- 12903690 TI - The status of women's health in the United States. PMID- 12903691 TI - Exercising during pregnancy. PMID- 12903692 TI - Hyperemesis gravidarum. PMID- 12903693 TI - Current screening instruments related to alcohol consumption in pregnancy and a proposed alternative method. AB - Because alcohol is a known teratogenic substance that negatively affects the fetus, screening for alcohol use is included in maternal child texts for nurses. They present no standard screening approach, however. In the general literature, a multitude of screening instruments exist for the purpose of detecting maternal alcohol dependence but few allow the nurse to detect fetal alcohol exposure in the absence of maternal alcohol dependence. After a comparative analysis of existing screening instruments, we concluded that the Timeline Followback method has the most utility as a screening instrument to detect fetal exposure to alcohol. The purpose of this article is to present a critical review of current screening instruments related to alcohol consumption in pregnant women and to propose the use of the Timeline Followback method as the preferred approach. PMID- 12903694 TI - Noise and the premature infant: physiological effects and practice implications. AB - Premature infants in the neonatal intensive-care unit often are exposed to continuous loud noise. This excess auditory stimulation creates negative physiological responses, such as increased heart and respiratory rates and decreased oxygen saturation. Modifying the neonatal intensive-care unit to reduce noise levels provides an environment more conducive to maturation and recovery from illness. This article reviews the literature addressing the pathophysiology and physiological changes of the premature infant in response to stress and discusses practice interventions for decreasing noise levels in the neonatal intensive-care unit. These interventions include installing sound absorbing materials and minimizing conversations of the staff. PMID- 12903695 TI - What we know about managing menopausal hot flashes: navigating without a compass. AB - Hot flashes and night sweats are frequently experienced as the cardinal symptoms of menopause. However, their physiological basis has not been explained; nor have any potential risks been explored. Current knowledge and theoretical perspectives regarding hot flashes will be presented and contrasted with evidence for an emerging hypothesis of altered brain glucose availability as the hot flash trigger. Perspectives regarding hormone therapy and alternative therapies for treatment of hot flashes will be presented and directions for future research reviewed. PMID- 12903696 TI - Transition to motherhood. AB - OBJECTIVE: To synthesize the results of nine qualitative studies related to the transition to motherhood, using Noblit and Hare's method of meta-synthesis development. DATA SOURCES: CINAHL, MEDLINE, PsycINFO, Sociological Abstracts, and Dissertation Abstracts, using the keywords transition, adaptation, motherhood, and mother. STUDY SELECTION: Studies were evaluated for inclusion related to study focus and comparability of findings. Nine studies were included in the final sample, four using a grounded theory methodology and five using either a phenomenologic or phenomenologic/hermeneutic method. DATA EXTRACTION: A series of overlapping, repeating steps as outlined by Noblit and Hare were followed in conducting this meta-synthesis. Detailed tables of metaphors, themes, concepts, and phrases from each study were constructed and studies were compared by means of reciprocal translations. DATA SYNTHESIS: Two processes inherent in maternal transition emerged: engagement and growth and transformation. In addition, five thematic categories signifying areas of disruption present in the maternal transition and 13 underlying themes were revealed. CONCLUSIONS: To facilitate maternal transition, nurses must remain sensitive to the maternal insecurity of primiparae and encourage their maximum maternal engagement, particularly during periods of forced mother-infant separation such as the newborn's admission to the intensive-care unit. In addition, prenatal discussion of realistic expectations for the transitional period, ongoing support through the first 6 months postpartum, and the use of role models should be integrated into postpartum support programs to assist mothers in negotiating typical areas of disruption during maternal transition. PMID- 12903697 TI - Postpartum women's perceptions of the hospital environment. AB - OBJECTIVE: To describe postpartum women's perceptions of the hospital environment and how it affects their postpartum experiences. METHOD: Descriptive study using content analysis of interview transcripts. SETTING: Participants were interviewed in their homes within the first 3 postpartum weeks about their postpartum experiences. PARTICIPANTS: Thirty-one primiparous women who gave birth vaginally to full-term newborns. They were recruited from two large childbirth education programs in the Pacific Northwest. RESULTS: The categories addressing the hospital environment were context, physical conditions, sociocultural conditions, contingencies, and consequences. Overall, these women had more negative perceptions than positive ones of the hospital environment. Women's perceptions seemed to be most influenced by the context of care. Women with mother/baby care had a greater proportion of positive perceptions than women with other care modalities. CONCLUSION: Nurses need to modify or change the hospital environment to promote more positive experiences for new mothers. PMID- 12903698 TI - Comparing the use of hydrogel dressings to lanolin ointment with lactating mothers. AB - OBJECTIVE: To evaluate the use of hydrogel dressings for the prevention and treatment of nipple soreness in lactating women as compared with the common intervention of lanolin ointment. The hypothesis was as follows: Participants using hydrogel dressings as a preventive measure for nipple soreness will experience greater pain relief and a lower rate of nipple wounds as compared with the control group. The secondary hypothesis was that the reduction of nipple soreness in the treatment group would produce a longer duration of breastfeeding as compared with the control group. DESIGN: A multicentered, prospective, randomized controlled clinical trial evaluating a sample of 106 lactating mothers. SETTING: Study sites were the University of Alabama Medical Center at Birmingham (an inner-city teaching hospital) and Northeast Health System (a community hospital in Beverly, Massachusetts). PARTICIPANTS: Participants were older than age 18, fluent in English, and had an operational telephone in the residence. Other inclusion criteria were singleton, vaginal deliveries; no prior breastfeeding experience; and written informed consent. INTERVENTIONS: Participants were randomized to either the lanolin ointment or the hydrogel dressings group and received instructions specific to their assignment. All participants received breastfeeding education provided by a board-certified lactation consultant. MAIN OUTCOME MEASURES: During the initial 12 study days, participants identified pain intensity using a numeric pain intensity scale and verbal descriptor scale. Subjective data were collected via self-reported skin assessments of the bilateral breasts, nipples, and areolae. Breastfeeding duration was established by a follow-up telephone call at 2 months. RESULTS: The hydrogel dressings group had significantly greater reduction in pain score mean values at baseline, on study Day 10, and on study Day 12 in comparison to the control group. Participants using the hydrogel dressings discontinued treatment sooner than participants in the lanolin ointment group. The lanolin ointment group had eight breast infections, whereas the hydrogel dressings group had none. CONCLUSION: Hydrogel dressings are a safe, available treatment that provided more effective pain management for nipple soreness than the common intervention of lanolin ointment. PMID- 12903699 TI - Use of a maternity support binder for relief of pregnancy-related back pain. AB - OBJECTIVE: To examine the feasibility, acceptability, and effectiveness of a support binder for low back pain in pregnancy. DESIGN: Pilot study, using a prospective, two-group design with repeated measures. SETTING: Ambulatory maternity clinic in a tertiary care teaching hospital. PARTICIPANTS: Women of at least 20 weeks gestation with low back pain, but no preexisting back or disc disease. Thirty women assigned to the intervention group and 10 to a comparison group. INTERVENTIONS: Participants completed a pain assessment at pretest. Intervention participants received a maternity support binder to wear while awake for 2 weeks. At an appointment 2 weeks later, a posttest questionnaire and a taped interview were administered. The comparison group participants received the support binder after the second appointment. MAIN OUTCOME MEASURES: Back pain intensity, duration, and effect on daily activities were assessed using a pain in pregnancy questionnaire. RESULTS: The intervention group had significant reduction in mean pain scores and effect of pain on daily activities, including family, house and yard, recreational, exercise, and sleep. Interaction of group by time was significant for change in pain and effect on family, house and yard, and exercise activities. CONCLUSION: The use of a support binder for pregnancy related low back pain is a promising intervention and was well-accepted by the participants. PMID- 12903700 TI - The effects of heparin versus normal saline for maintenance of peripheral intravenous locks in pregnant women. AB - OBJECTIVE: To compare the efficacy of two available preparations (heparin, 10 U/mL, 1 mL, vs. normal saline, 1 mL) used for maintaining patency in peripheral intravenous (IV) locks during pregnancy. DESIGN: Prospective, randomized, and double-blind. Eligible patients who were to receive a peripheral intermittent IV lock were randomly assigned to receive either heparin flushes or normal saline flushes for IV lock maintenance. IV locks were flushed after each medication administration, or at least every 24 hours, with the assigned blinded flush solution. Intermittent IV lock sites were also evaluated every 12 hours for the development of phlebitis. SETTING: A large academic medical center in the Midwest that has both community-based and regional-referral obstetric practices with more than 2,000 deliveries per year. PARTICIPANTS: A convenience sample included 73 hospitalized pregnant women who were between 24 and 42 weeks gestation. Exclusions from the study were women with significant abnormalities in the fetal heart tracing on admission, cervical dilation > 4 cm, presence of hypersensitivity to heparin, presence of clotting abnormalities, and anticoagulation therapy (including low-dose aspirin). RESULTS: Data indicate there were no statistically significant differences in IV lock patency nor in phlebitis between heparin or normal saline flushes. CONCLUSIONS: This study provides support that both normal saline and heparin in the doses studied may be equally effective in the maintenance of peripheral IV locks. Due to small sample size, additional studies are needed to determine optimal therapy over time. PMID- 12903701 TI - Resources for evidence-based practice, July-August 2003. PMID- 12903702 TI - Full circle: the evolution of preconception health promotion in America. AB - Preconception health promotion encourages and enables a woman to actively plan pregnancy, enter pregnancy in good health with as few risk factors as possible, and be informed about her general and reproductive health before conception. Nurses have provided this holistic health care since the early 1900s. In succeeding decades, various components of and methods for promoting preconception health have been discovered and emphasized. Embracing preconception health promotion as wellness for life brings us full circle. PMID- 12903704 TI - Interconceptional counseling after perinatal and infant loss. AB - Historically, preconceptional health promotion has been recommended for all prospective parents to improve perinatal outcomes. Preconceptional health promotion and interconceptional counseling may be even more beneficial for parents who have had previous perinatal losses. Perinatal loss can be devastating, with long-term effects on subsequent pregnancies and children. A theoretical framework for interconceptional counseling after perinatal loss needs to be developed. Interconceptional counseling can give couples important information to improve outcomes, acknowledge fears and anxieties, evaluate genetic risks, facilitate grieving, and explore attachment and parenting issues. PMID- 12903703 TI - Preconception health counseling for women exposed to teratogens: the role of the nurse. AB - Women with unintended pregnancies who are exposed to teratogens constitute the highest risk group for fetal harm. Teratogen exposures come from substances, medications, chronic and acute diseases, and environmental factors. Nurses play a critical role in reducing unintended pregnancy and promoting preconception health. A greater understanding of the role of teratogens and strategies to improve history taking and help women prevent unintended pregnancy will improve nurses' ability to reduce teratogen exposure in women at risk. PMID- 12903705 TI - Changing provider practices to enhance preconceptional wellness. AB - Primary care providers who fail to provide preconceptional care to every woman of reproductive age during each primary care visit are losing key preventive opportunities. Changing provider practices involves multidisciplinary input into the planning of care, support of clinical leaders and professional organizations, and adequacy of funding for proposed changes. Programs in California, North Carolina, and South Carolina have demonstrated significant changes in provider acceptance of preconceptional. PMID- 12903706 TI - Preconceptional wellness as a routine objective for women's health care: an integrative strategy. AB - Preconceptional health promotion should not be approached as an isolated activity. Instead, a new approach to women's wellness is needed. Rather than targeting care to women based on their pregnancy status or desires, health promotion and disease prevention should be integrated into a continuum of care throughout the life cycle. When care for women is viewed as an integrated continuum approach to health, rather than as a series of episodic events, higher levels of women's wellness will be achieved. This approach is likely to result in healthier women, pregnancies, and offspring. These outcomes are consistent with the goals of preconceptional health promotion. Using several case illustrations, this article highlights the benefits of integrating care into a continuum model. PMID- 12903707 TI - Tissue engineering: confronting the transplantation crisis. PMID- 12903708 TI - Human embryonic stem cells--realising the potential. PMID- 12903709 TI - Human embryonic or adult stem cells: an overview on ethics and perspectives for tissue engineering. AB - Over the past few years, research on animal and human stem cells has experienced tremendous advances which are almost daily loudly revealed to the public on the front-page of newspapers. The reason for such an enthusiasm over stem cells is that they could be used to cure patients suffering from spontaneous or injuries related diseases that are due to particular types of cells functioning incorrectly, such as cardiomyopathy, diabetes mellitus, osteoporosis, cancers, Parkinson's disease, spinal cord injuries or genetic abnormalities. Currently, these diseases have slightly or non-efficient treatment options, and millions of people around the world are desperately waiting to be cured. Even if not any person with one of these diseases could potentially benefit from stem cell therapy, the new concept of "regenerative medicine" is unprecedented since it involves the regeneration of normal cells, tissues and organs which could allow to treat a patient whereby both, the immediate problem would be corrected and the normal physiological processes restored, without any need for subsequent drugs. However, conflicting ethical controversies surround this new medicine approach, inside and outside the medical community, especially when human embryonic stem cells (h-ESCs) are concerned. This ethical debate on clinical use of h-ESCs has recently encouraged the research on "adult" stem cells (ASCs) regarded as a less conflicting alternative for the future of regenerative medicine. PMID- 12903710 TI - Stem cell biology and plasticity. PMID- 12903711 TI - Liver tissue engineering: successes & limitations. PMID- 12903712 TI - Pancreatic islet and stem cell transplantation in diabetes mellitus: results and perspectives. PMID- 12903713 TI - Cell-based therapy of chronic degenerative diseases of the central nervous system. AB - The traditional methods of pharmacotherapy of the degenerative diseases of the central nervous system do not frequently allow one to achieve the desired clinical effect. The fundamentally new approach for the treatment of severe neurological diseases is provided by the methods of biological medicine, in particular, transplantation of a complex of fetal tissues. Cell-based therapy was used to treat patients with multiple sclerosis; ante-, intra- and postnatal lesions; consequences of hemorrhagic and ischemic apoplexies; neuritis of facial nerve; sclerosis; Parkinson's disease; Alzheimer's disease; epilepsy and other types of pathologic process. The source material for obtaining a suspension of cells was the fetuses of allogenic origin. The suspension of brain cells in amounts of up to 1.5 x 10(8) cells and vitality not less than 40% was administered to the patients into liquor spaces using the method of endolumbar puncture. The total number of transplantations was 1900. Practically in all the cases FT was tolerated well. Positive clinical and immunologic changes were observed in the majority of the patients, thus, remission induction (in the patients with the progressive course of multiple sclerosis) for a period over 12 months was registered in 87.5% of the cases. Noteworthy that considerable changes were observed in immunograms: depression of antibody levels to brain-specific proteins, native and denatured DNA; quantitative and qualitative improvement of lymphocyte subpopulation indices, positive changes in the immunoregulatory index. Clinically, in 69% of the cases there was an improvement in more than one neurological defect and a change in the values of the Kurtzke scale towards a decrease by 2-3 points. The conduct of cell therapy with the MS patients under the acute process conditions after liquorosorption allowed the arresting of clinical manifestations and the creation of preconditions for further restoration. The retrobulbar transplantations provided a quick arrest of the retrobulbar neuritis clinical symptoms and in one case an almost complete restoration of vision in the patient with amaurosis (blindness). The remission duration has a marked direct dependence on the number of courses of endolumbar transplantations. Thus, the method of cell therapy with the use of human tissue transplantations is safe and can be used for different neurodegenerative lesions of the central nervous system. The high efficacy of the method suggests the possibility and necessity of using this method as an alternative of classical pharmacological therapy. An important element of cell therapy is the control after the state of the patient's immunity system. PMID- 12903714 TI - CD34+ cells in hematopoietic stem cell transplantation. PMID- 12903715 TI - The effects of different growth factors on human bone marrow stromal cells differentiating into hepatocyte-like cells. PMID- 12903716 TI - Oxidative DNA damage biomarkers used in tissue engineered skin. PMID- 12903717 TI - Biomarkers used to detect genetic damage in tissue engineered skin. PMID- 12903718 TI - Design peptide scaffolds for regenerative medicine. PMID- 12903719 TI - Progresses in synthetic vascular prostheses: toward the endothelialization. PMID- 12903720 TI - Adhesion and growth of rat aortic smooth muscle cells on lactide-based polymers. AB - Biodegradable materials based on polymers of hydroxy acids are studied for application in artificial vascular substitutes. Polymers with functional surfaces are being developed, carrying specific recognition structures to affect selectively the adhesion and proliferation of endothelial cells (EC) and vascular smooth muscle cells (VSMC). This preliminary study focuses on evaluation of adhesion and growth of VSMC on surfaces of polylactide polymers and those modified by amphiphilic polylactide/poly(ethylene oxide) copolymers. Poly(L lactic acid), PLLA, and poly(DL-lactic acid), PDLLA, and a block copolymer of lactide with a carboxylated poly(ethylene oxide) segment, PLLA-b-PEO-COOH, were synthesized by controlled polymerization of L and D,L-lactide, respectively, and using delta-hydroxy-Z-carboxymethyl-PEO as a macroinitiator for the copolymer. Films of polymers were deposited on glass coverslips by a spin-coating method. Uncoated glass coverslips and Falcon dishes were used as control substrates. VSMC were obtained from the thoracic aorta of young adult male Wistar rats by explantation method and seeded in Dulbecco-Modified Eagle MEM with 10% foetal bovine serum. The number of adhering cells, their shape, size of cell-material contact area and cell population doubling time were evaluated from day 1 to 7 after seeding. It was found that both PLLA and especially PDLLA relatively well supported adhesion and growth of VSMC. However, on carboxylated surfaces of the PLLA-b-PEO-COOH copolymer, a lower number of initially adhering cells (by 37% than on Falcon dishes, pdelta0.05), smaller cell spreading area (by 45% and 37% than on glass and Falcon dishes, respectively, pdelta0.01) and longer doubling time (by 49% and 31% than on glass and Falcon dishes, pdelta0.001). Thus, surfaces coated by a PLA/PEO-COOH copolymer can be used as minimum background surface to reveal the effect of other more specific adhesion structures. PMID- 12903722 TI - Polymer based scaffolds and carriers for bioactive agents from different natural origin materials. PMID- 12903721 TI - Biodegradable copolymers carrying cell-adhesion peptide sequences. AB - Amphiphilic block copolymers are used to create bioactive surfaces on biodegradable polymer scaffolds for tissue engineering. Cell-selective biomaterials can be prepared using copolymers containing peptide sequences derived from extracellular-matrix proteins (ECM). Here we discuss alternative ways for preparation of amphiphilic block copolymers composed of hydrophobic polylactide (PLA) and hydrophilic poly(ethylene oxide) (PEO) blocks with cell adhesion peptide sequences. Copolymers PLA-b-PEO were prepared by a living polymerisation of lactide in dioxane with tin(II)2-ethylhexanoate as a catalyst. The following approaches for incorporation of peptides into copolymers were elaborated. (a) First, a side-chain protected Gly-Arg-Gly-Asp-Ser-Gly (GRGDSG) peptide was prepared by solid-phase peptide synthesis (SPPS) and then coupled with delta-hydroxy-Z-amino-PEO in solution. In the second step, the PLA block was grafted to it via a controlled polymerisation of lactide initiated by the hydroxy end-groups of PEO in the side-chain-protected GRGDSG-PEO. Deprotection of the peptide yielded a GRGDSG-b-PEO-b-PLA copolymer, with the peptide attached through its C-end. (b) A protected GRGDSG peptide was built up on a polymer resin and coupled with Z-carboxy-PEO using a solid-phase approach. After cleavage of the delta-hydroxy-PEO-GRGDSG copolymer from the resin, polymerisation of lactide followed by deprotection of the peptide yielded a PLA-b-PEO-b-GRGDSG block copolymer, in which the peptide is linked through its N-terminus. PMID- 12903723 TI - Preparation and characterization of natural/synthetic hybrid scaffolds. PMID- 12903724 TI - Polyhipe polymer: a novel scaffold for in vitro bone tissue engineering. PMID- 12903725 TI - Pancreatic islet culture and transplantation using chitosan and PLGA scaffolds. PMID- 12903726 TI - Challenges and prospects for targeted transgenesis in livestock. Practical applications of gene targeting. PMID- 12903727 TI - Controlled release of bioactive agents in gene therapy and tissue engineering. PMID- 12903728 TI - Interleukin 1 (IL-1) induces the activation of Stat3. PMID- 12903729 TI - The use of antibodies in diagnosis and therapy of cancer. PMID- 12903730 TI - Detection of phage displayed peptides with blocking ability in vascular endothelial growth factor (VEGF) model. PMID- 12903731 TI - Induced production of rabbit articular cartilage-derived chondrocyte collagen II on polyhydroxyalkanoate blends. AB - Three-dimensional porous scaffolds made from blend polyesters of polyhydroxyalkanoates (PHA) were evaluated with respect to their effects on collagen II production. Semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR) was used to assay collagen II mRNA for evaluation of the ability for the blend scaffolds to induce collagen II production. It was found that scaffold of poly-3-hydroxybutyrate (PHB) blended with a copolyester consisting of 3-hydoxybutyrate-co3-hydroxyhexanate (PHBHHx) in a radio of 1:2 had strong ability to induce collagen II production compared with other ratios of PHB/PHBHHx and poly-L-lactic acid (PLA). Collagen II expression level remained approx. 1.5-times higher on the above-mentioned blend compared with that of PLA throughout four weeks cultivation. At the same time, the PHB/PHBHHx blend (1:2) exhibited a similar tendency as PLA to maintain the collagen 11 expression level over the entire culture period. The results showed that the PHB/PHBHHx blend (1:2) reported to improve mechanical properties compared with PHB or PHBHHx improved biocompatibility in comparison with PHB, PHBHHx, PLA and other blending systems consisting of PHB/PHBHHX in ratios of 2:1 and 1:1, respectively. PMID- 12903732 TI - Development of a cultured dermal substitute composed of a spongy matrix of hyaluronic acid and atelo-collagen combined with fibroblasts: fundamental evaluation. AB - We have developed an allogeneic cultured dermal substitute (CDS) by cultivating fibroblasts on a 2-layered spongy matrix of hyaluronic acid (HA) and atelo collagen (Col). The HA sponge was designed to have a honeycomb structure with many holes (0.5 mm diameter) separated by a distance of 4 mm. Part of the Col sponge was able to penetrate into these holes, and the resulting anchoring structure allows binding of a HA spongy layer with a Col spongy layer. The preparation of the CDS consists of two steps: (i) attachment of cells to the Col surface of the hydrated 2-layered spongy matrix and (ii) proliferation of cells on this sponge immersed in culture medium. The aim of the present study was to assess properties of fresh and cryopreserved CDS. Fibroblasts seeded on the Col surface of the 2-layered spongy matrix attached, proliferated and released vascular endothelial growth factor (VEGF) and fibronectin. The amount of VEGF released from cryopreserved CDS after thawing slowly in an incubator at 37 degrees C and re-cultivation for 1 week was about 300 pg/ml. After thawing quickly in a water bath at 37 degrees C and re-cultivation for 1 week, the amount of VEGF released was about 600 pg/ml. These findings indicate that the cryopreserved CDS maintained its ability to release a significant amount of VEGF. Retention of the therapeutic properties of CDS after cryopreservation is important for clinical use. PMID- 12903734 TI - Cell-compatible properties of calcium carbonates and hydroxyapatite deposited on ultrathin poly(vinyl alcohol)-coated polyethylene films. AB - Poly(vinyl alcohol) (PVA) was coated onto polyethylene (PE) films by a repetitive adsorption and drying process, and then the PVA-coated PE films were alternately immersed into aqueous solutions of Ca2+ and CO3(2-) ions (alternate soaking cycles), to deposit calcium carbonate (CaCO3) onto the films. The PVA coating was essential for the CaCO3 deposition. The amount of CaCO3 deposited increased with an increasing number of cycles. Scanning electron microscopic observations and attenuated total reflection spectra revealed the presence of both calcite and aragonite as the crystal structures of CaCO3 on the film. L929 fibroblast cells adhered and proliferated on these CaCO3-deposited PE films, as well as the hydroxyapatite-coated PE films previously prepared. It was found that the PVA coating and the subsequent deposition of calcium salts on certain films facilitated cell compatibility. PMID- 12903733 TI - Modification of porous aminopropyl-silicate microcapsule membrane by electrically bonded external anionic polymers. AB - Biocompatibility and permeability of a microcapsule membrane governs the function of a microcapsule-shaped bioartificial pancreas. We have previously developed an alginate/sol-gel-synthesized aminopropyl-silicate/alginate microcapsule (Alg/AS/Alg), which had insufficient biocompatibility. The purpose of this study was to investigate whether the biocompatibility could improve by modifying the external surface with other anionic polymers and to investigate an influence of the modification on the permeability of the membrane. Four kinds of anionic polymers, poly(oxyethylene)diglycolic acid (3 kDa), heparin (15 kDa), Alg (54 kDa) and carboxymethylcellulose (CMC, 60 kDa) were used as the external anionic polymers. The heparin-bonded gel bead had the largest resistance to the diffusion of small molecules. The molecular mass cut-off point of 150 kDa required for immunoisolation was maintained for all anionic polymers. Cellular overgrowth to the implanted islet-enclosing microcapsule, a sign of insufficient biocompatibility, was suppressed by altering the external surface material from Alg to CMC. These results suggest that the biocompatibility of the Alg/AS/anionic polymer membrane can be improved by using a biocompatible anionic polymer. At the same time, it is suggested the influence on the permeability has to be investigated to develop an optimal microcapsule for bioartificial pancreas. PMID- 12903735 TI - Mechanical properties and in vitro degradation of self-reinforced radiopaque bioresorbable polylactide fibres. AB - The aim of this study was to evaluate the effect of the radiopaque filler, barium sulfate (BaSO4), on the mechanical properties of self-reinforced bioresorbable fibres. The bioresorbable polymer was a copolymer of L- and D-lactide with an L/D monomer ratio of 96:4 (96L/4D PLA). The fibres were manufactured using an extrusion and a drawing process. Three different methods of processing the composites were studied. The materials were blended prior to extrusion. In the first method, the BaSO4 powder was mixed with the polymer granulates by hand (manual blending). The blend was then processed using a twin-screw extruder. The second and third methods utilized a single-screw extruder. In the second method, the BaSO4 powder was manually mixed with the polymer prior to extrusion. In the third method, the BaSO4 powder was mechanically attached on the polymer granulates (mechanical blending) prior to extrusion. The mechanical and chemical properties of the radiopaque bioresorbable fibres were measured after processing and during in vitro degradation. The fibres were gamma, plasma or EtO sterilized. There was no statistical difference in the mechanical properties of the fibres when manufactured using the twin-screw extrusion with manual blending or the single-screw extrusion with mechanical blending. The gamma sterilization markedly decreased the initial intrinsic viscosity of all fibres, whereas the plasma and EtO sterilization methods had no effect on the initial intrinsic viscosity. During in vitro testing, the loss in the intrinsic viscosity occurred at the same rate whether the fibres were loaded with the barium sulfate or not. PMID- 12903736 TI - Theoretical and experimental evaluation of the radial force of self-expanding braided bioabsorbable stents. AB - This study was carried out to evaluate if the analytical model developed by Jedwab and Clerc for calculating the mechanical properties of metallic braided stents is also valid for bioabsorbable braided stents. An analytical model could be used to shorten the development cycle of stents by reducing the amount of in vitro testing. Jedwab and Clerc derived formulae for longitudinal stiffness and radial pressure stiffness. The longitudinal stiffness was defined by measuring the stent elongation under load. The radial pressure stiffness was defined from the slope of the load-displacement curve measured with the testing method described by Agrawal and Clark where a collar is placed around the stent to compress it. The radial pressure stiffness was measured with and without lubrication to evaluate the effects of friction between the stent and collar and in the stent structure itself. Two bioabsorbable braided stents and one metallic braided stent were used in the measurements. The metal stent test results were consistent with what was reported by Jedwab and Clerc. However, the analytical model was not applicable to bioabsorbable stents. This was mainly due to the larger fibre diameter of the bioabsorbable stents, which prevents the fibres from freely collapsing when the stent diameter decreases. The analytical model is based on an assumption that the fibres behave independently. However, the testing method described by Agrawal and Clark provided a useful tool to compare the radial force of self-expanding stents. PMID- 12903737 TI - Biocompatible collagen scaffolds from a human amniotic membrane: physicochemical and in vitro culture characteristics. AB - A reconstituted collagen membrane from human amnion has been investigated as a source of collagen matrix, which could be used as a substratum for culturing human fibroblasts. The suitability of pepsin-solubilized reconstituted human amniotic membrane, before and after cross-linking with chitosan, as a dermal matrix for culturing fibroblast was assessed by morphologic, physicochemical, cytotoxic and histochemical methods. Measurement of thermodynamic behaviour, by differential scanning calorimetric (DSC) and thermogravimetric analysis (TGA), and tensile strength suggested that the cross-linked membrane had sufficient elasticity to serve as an efficient dermal substrate for in vitro culture of fibroblasts. Fibroblasts cultured on the chitosan cross-linked collagen membrane had good adherence, retaining their morphology as indicated by microscopic analysis. Proliferation of fibroblasts. observed on this membrane affirms its non toxic nature. These results support the application of reconstituted human amniotic collagen membrane as collagenous scaffolds to culture fibroblasts in vitro. PMID- 12903738 TI - Synthetic studies on nonthrombogenic biomaterials 14: synthesis and characterization of poly(ether-urethane) bearing a Zwitterionic structure of phosphorylcholine on the surface. AB - A new Zwitterionic compound of the phosphorylcholine analogue, 4-hydroxylbutyl phosphorylcholine (HBPC), was synthesized and characterized. HBPC was chemically tethered onto the surface of poly(ether-urethane) (PEU) films with hexamethylene diisocyanate (HDI) as a coupling agent. The existence of a phosphorylcholine structure on the PEU surface was demomstrated by attenuated total reflection Fourier transform infrared spectroscopy (ATR-FT-IR), X-ray photoelectron spectroscopy (XPS) and water contact angle measurements. The nonthrombogenicity of the modified films was evaluated by platelet-rich plasma (PRP) assay. The results showed that films grafted with HBPC have excellent platelet adhesion resistance. PMID- 12903739 TI - Hydrogel-coated textile scaffolds as candidate in liver tissue engineering: II. Evaluation of spheroid formation and viability of hepatocytes. AB - In this study we evaluate the performance of primary rat hepatocytes and HepG2 cells on chitosan-collagen hydrogel-coated textile scaffolds. Light microscopy and electron microscopic observations showed attachment and aggregate formation tendency of hepatocytes on the scaffolds. As tested by the tetrazolium reduction (MTT) assay it was evident that cells had preserved mitochondrial functionality. It was also observed that pure collagen and collagen blended scaffolds allowed higher cell growth than pure chitosan scaffold. Fluorescent live/dead staining showed a metabolically active, viable cell population on all scaffold compositions with occurrence of few dead cells. Cell functionality was confirmed by secretion of albumin, which was maintained throughout culture period. Take collectively our results suggests that hydrogel-coated textile scaffolds could be promising for tissue-engineering applications, as they allow favorable hepatocyte attachment, spheroid formation and maintenance of function. These scaffolds could be useful for co-culturing hepatocytes and non-parenchymal endothelial cells in bioartificial liver support systems. PMID- 12903740 TI - The efficiency of titanocene as photoinitiator in the polymerization of dental formulations. AB - A comparative kinetic study of the polymerization of tetraethyleneglycol dimethacrylate (TEEGDM) under visible light irradiation was carried out in order to determine the effectiveness of titanocene as photoinitiator in this process. The pair camphorquinone/dimethylaminoethyl methacrylate, CQ/DMAEMA (0.5/0.5%, by weight), was used as a reference photoinitiatior system. The reactions were carried out both in the presence and absence of oxygen (in N2) and with variable and constant titanocene concentration and incident light intensity. It was observed that in both atmospheres the polymerization reaction commenced rapidly and its initial rate was comparable with that obtained in the presence of the classic initiation system. However, after reacting for a few seconds this process suffered abrupt deactivation, this tendency being more noticeable at low titanocene concentrations. To explain the observed kinetic behavior, the study of the photofragmentation of titanocene upon irradiation with visible light in both atmospheres was carried out. The quantum yield calculated in the presence of O2 (phifr = 8 at 465 nm) suggests the existence of a chain reaction with participation of oxygen, and this explains, in turn, the high value of the initial rate of polymerization and also the rapid disappearance of the photoinitiator. In an inert atmosphere the photofragmentation occurs in a conventional way, this being a very effective process with quantum yield of 0.7 at 465 nm. However, it seems that only a small part of the active species acts as initiating radicals and these are not sufficient to achieve the appropriate monomer conversion. PMID- 12903741 TI - Ribosomal protein P0 from Aedes albopictus mosquito cells: cDNA cloning and analysis of expression. AB - Ribosomal protein P0 is one of the highly conserved phosphorylated proteins in the large subunit of eukaryotic ribosomes. P0 has been shown in Drosophila to be a multifunctional protein that associates with elongation factor eEF2 to facilitate translation and also plays a role in DNA repair. In this paper we describe the cloning and characterization of the full-length cDNA encoding P0 from the mosquito, Aedes albopictus. In vitro translation showed that the cDNA encoded a 34kDa protein that corresponded in size to a phosphorylated protein from the large ribosomal subunit after in vivo labeling with [32P]orthophosphate. When cells were exposed to various stresses, expression of P0 appeared to differ from that of rpL34. Preliminary RNAi experiments suggested that downregulation of P0 is correlated with apoptosis in C7-10 mosquito cells. PMID- 12903742 TI - Inter and intra-specific hybridization in tuco-tucos (Ctenomys) from Brazilian coastal plains (Rodentia: Ctenomyidae). AB - The present work describes chromosomal polymorphisms in zones of contact between divergent populations of Ctenomys minutus parapatrically distributed in the coastal plain of southern Brazil, and inter-specific hybridization with C. lami a closely related species. A sample of 171 specimens from 32 sample sites distributed along 161 km of the coastal plain was cytogenetically analyzed. Nine polymorphic populations were found: four with specimens with 2n = 46-48 (autosomal arm number (AN) = 76); three only have specimens with 2n = 47 and 48; one population sampled presented specimens with 2n = 43-46 (AN = 74-76) and one population with 2n = 50-52 (AN = 76-80). The remainder populations were fixed for 2n = 42, 46 or 48. The variation is the result of Robertsonian mechanisms of chromosomal evolution and a fusion in tandem rearrangement. The polymorphisms have been considered the result of secondary contact of populations after divergence in allopatry. The geomorphological evolution of the coastal plain provides clues to the possible existence of past geographic barriers acting over populations of Ctenomys, during the Holocene. PMID- 12903744 TI - Contrasting allozyme and RAPD variation in spider populations from patchy forest habitats. AB - During the last 1000 years, massive deforestation events have occurred in Flanders (the northern part of Belgium) and the remaining forests have become very isolated patches. It is expected that organisms bound to these patchy forest habitats and with limited dispersal capacities will likely experience strong effects of genetic drift. One such organism is the spider Coelotes terrestris. Allozyme data suggested that 10 Flemish populations of this spider showed little genetic variation, as only one out of 20 loci was polymorphic (phosphoglucose isomerase). In view of this result, we used random amplified polymorphic DNA (RAPD) markers to test whether this lack of allozyme diversity is an inherent feature of the populations and/or species studied or whether it rather reflects a characteristic of the markers and/or methods used. Since the RAPD data revealed a substantial amount of genetic diversity in the same 10 populations, our results suggest that the latter is true. Furthermore, the RAPD data agree with the expectations for an organism with low dispersal capacities that has lived in isolated forest patches for at least 200 generations. Supplemented with the results of other techniques and studies, these findings might be of importance for the future conservation of this spider species in Flanders. PMID- 12903743 TI - Molecular evolution of tephritid fruit flies in the genus Bactrocera based on the cytochrome oxidase I gene. AB - Fruit flies of the genus Bactrocera (Diptera: Tephritidae) are one of the major economically important insects in Asia and Australia. Little attention has been given to analyses of molecular phylogenetic relationships among Bactrocera subgenera. By using mitochondrial cytochrome oxidase I gene (COI) sequences, the phylogenetic relationships among four subgenera, Asiadacus, Bactrocera, Hemigymnodacus, and Zeugodacus, were investigated. Nucleotide diversity within subgenera ranged from 11.7 to 12.4%, and the net divergence among subgenera ranged from 11.2 to 15.7%. Phylogenetic trees calculated from both maximum parsimony and neighbor-joining phylogenetic analysis methods were highly congruent in terms of tree topologies. Phylogenetic analysis of mitochondrial COI sequences suggests that tephritid fruit fly species, which attack cucurbit plants, that is, Asiadacus, Hemigymnodacus and Zeugodacus, were more closely related to each other than to fruit fly species of the subgenus Bactrocera, which attack plants of numerous families. Our data supports previous classification of Bactrocera based on morphological characters. However, the phylogenetic tree showed the polyphyletic of fruit flies in subgenus Zeugodacus. Possible causes of speciation among fruit flies species in this genus were also discussed. PMID- 12903745 TI - Effects of genetic sex and genomic background on epistasis in rainbow trout (Oncorhynchus mykiss). AB - Epistasis among quantitative trait loci (QTL) for survival (upper thermal tolerance, UTT) and morphological (fork length, FL and condition factor, K) traits was detected in purestrain and interstrain rainbow trout (Oncorhynchus mykiss) families. One sex-linked (OmyFGT19TUF) and three autosomal (Omy325UoG, Ssa14DU and Ssa20.19NUIG; linkage groups B, D and S, respectively) microsatellite loci linked to UTT QTL in this species were used. Within half sib families, significant effects of full sib family on epistasis involving Omy325UoG and OmyFGT19TUF were detected at a rate significantly higher than expected for UTT (p < 0.001*) and FL (p < 0.01*), using results significant at comparisonwise significance thresholds derived from permutational analysis. Measured across half sib families, the phenotype of female genotypic classes was more divergent from the family trait mean than that of males where epistasis involved the sex-linked locus OmyFGT19TUF (p = 0.0176*), and also for means over all families (p = 0.0355*). Female genotypic classes were also more divergent (p = 0.0011 **) from the full sib trait mean where three-way interaction between OmyFGT19TUF, one of the autosomal loci and full sib family was significant, and marginally more divergent for trait means of genotypic classes across all full sib families (p = 0.0856+). There was no evidence that these effects were more pronounced in hybrid F1 families than purestrains. PMID- 12903746 TI - Replication banding patterns in the spined loach, Cobitis taenia L. (Pisces, Cobitidae). AB - The chromosomal complement of Cobitis taenia was analysed by replication banding techniques to determine whether there were specific patterns that could allow distinction of the different chromosomes. The diploid chromosome number of 2n = 48 is diagnostic of this species. In vivo 5-bromodeoxyuridine (5-BrdU) incorporation induced highly reproducible replication bands. Most of the chromosome pairs were distinguishable on the base of their banding patterns. The karyotype, consisting of five pairs of metacentrics, nine pairs of submetacentrics and 10 pairs of subtelocentrics and acrocentrics, was confirmed. C-banding and replication banding patterns were compared, and heterochromatin was both early and later replicating. C-positive heterochromatin in centromeric regions was mainly early replicating, but that located in pericentromeric regions was late replicating. Most of the late-replicating regions found interstitially were C-band negative. The results obtained so far for combined chromosomal staining methods of C. taenia and other Cobitis fish species are discussed. PMID- 12903747 TI - Allele frequency and genotype distribution of polymorphisms within disease related genes is influenced by ethnic population sub-structuring in Sudan. AB - Four single nucleotide polymorphisms (SNPs) and a variable number of tandem repeats (VNTR) polymorphism located within disease associated/causing genes were typed in four populations of different tribal and ethnic affiliation from the Sudan. The genotype and allele frequencies were compared with those of other groups from published and unpublished data of world populations. The combined Sudanese sample conformed with Hardy-Weinberg equilibrium (HWE) expectation. However, population sub-structuring according to ethnic/linguistic group indicated at least two SNPs in departure from HWE. Differences in allele frequencies and genotype distribution between groups was also noted in three of the four SNPs. The other loci were distributed homogeneously within the populations studied with genotype frequencies in agreement with HWE expectation. These results highlight the importance of inter-population stratification for polymorphic markers, as well as the potential influence of evolutionary history and ethnic variation of loci, in the general distribution of SNPs and other polymorphisms. PMID- 12903748 TI - Nucleolar organizing regions in Crassostrea angulata: chromosomal location and polymorphism. AB - Specimens of Crassostrea angulata collected from several natural populations, with or without metal contamination, located along the Southwestern coast of Spain were cytogenetically analyzed. The diploid number was 20 and all 10 chromosome pairs were metacentric. The nucleolar organizing regions (NORs) were identified on the telomeric region of the chromosome pair 10 by several methods, including silver nitrate staining (AgNO3), chromomycin A3 staining (CMA3) and fluorescence in situ hybridization (FISH). One or two primary Ag-NORs in the metaphase cells were most frequently found. Very infrequently, one or two secondary NORs were seen. High Ag-NOR and rDNA polymorphisms in both the size and/or the number occurred intra-individually, inter-individually and inter populationally. However some population-specific Ag-NOR polymorphisms were found to reach or be approaching to a level of significant difference. A comparison of number and frequencies between FISH-rDNA signals and Ag-NORs showed that some NORs were not transcriptionally active. Moreover, presence of pollutants on the medium could be favoring the presence of two NORs per cell more than acting modifying expression on the NORs. PMID- 12903749 TI - A method for marker-assisted selection based on QTLs with epistatic effects. AB - A method for marker-assisted selection (MAS) based on quantitative trait loci (QTLs) with epistatic effects is proposed. The efficiency of such method is investigated by simulations under a wide range of situations. In the presence of epistasis, MAS generally yields longer persistence response than that based exclusively on additive or additive and dominance. Neglecting epistasis could result in considerable loss in response, and more pronounced at later generations. In addition to population size and trait heritability, genetic variance configurations play an important role in determining both the short- and long-term efficiencies of MAS. MAS using breeding values not only achieves higher response, but also tends to have smaller standard error than other methods in most cases. Errors in QTL detection cause distinct reductions in responses to MAS in most cases. It is thus concluded that verifications of putative QTL and its magnitude of effect and accurate map chromosome location are imperative to realize the potentials of MAS. PMID- 12903750 TI - Analysis of heterochromatin by combination of C-banding and CMA3 and DAPI staining in two fish species (Pimelodidae, Siluriformes). AB - The chromosomes of Steindachneridion sp. (2n = 56) and Rhamdia quelen (2n = 58) were analyzed by C-banding (CB) and Chromomycin A3 (CMA3) and 4,6-diamidino-2 phenylindole (DAPI) staining, separately and consecutively, in order to understand the role of base-specific fluorochrome treatment after CB. Both species' chromosomes shared common staining profiles as follows. CB with Giemsa (CBG) revealed weak heterochromatic blocks in the telomeric regions of some chromosomes and conspicuous bands on the short arms of one chromosome pair, where nucleolar organizer regions (NORs) were evidenced by silver-staining. Without CB pretreatment, the NORs were stained conspicuously with CMA3, but not with DAPI. The latter uniformly stained all chromosomes, but leaving the NORs pale. Combination of CMA3 or DAPI staining with CB showed distinctive fluorescent blocks in the NOR-bearing short arms of the single chromosome pair along with several bright fluorescent signals on other chromosomes, which were not evidenced by single CMA3 or DAPI staining. These results suggest a modification of chromatin structure by CB treatment, which may increase the stainability of CMA3 and DAPI. PMID- 12903751 TI - Genetic divergence between Cyprinus carpio carpio and Cyprinus carpio haematopterus as assessed by mitochondrial DNA analysis, with emphasis on origin of European domestic carp. AB - Although common carp is the major fish species in Asian and European aquaculture and many domestic varieties have occurred, there is a controversy about the origination of European domestic common carp. Some scientists affirmed that the ancestor of European domestic common carp was Danube River wild common carp, but others considered it might be Asian common carp. For elucidating origination of European domestic common carp, we chose two representative European domestic common carp strains (German mirror carp and Russian scattered scaled mirror carp) and one wild common carp strain of Cyprinus carpio carpio subspecies (Volga River wild common carp) and two Asian common carp strains, the Yangtze River wild common carp (Cyprinus carpio haematopterus) and traditionally domestic Xingguo red common carp, as experimental materials. ND5-ND6 and D-loop segments of mitochondrial DNA were amplified by polymerase chain reaction and analyzed through restriction fragment length polymorphism (RFLP) and sequencing respectively. The results revealed that HaeIII and DdeI digestion patterns of ND5 ND6 segment and sequences of control region were different between European subspecies C. carpio carpio and Asian subspecies C. carpio haematopterus. Phylogenetic analysis showed that German mirror carp and Russian scattered scaled mirror carp belonged to two subspecies, C. carpio carpio and C. carpio haematopterus, respectively. Therefore, there were different ancestors for domestic carp in Europe: German mirror carp was domesticated from European subspecies C. carpio carpio and Russian scattered scaled mirror carp originated from Asian subspecies C. carpio haematopterus. PMID- 12903752 TI - FISH-mapping of 18S ribosomal RNA genes and telomeric sequences in the Japanese bitterlings Rhodeus ocellatus kurumeus and Tanakia limbata (Pisces, Cyprinidae) reveals significant cytogenetic differences in morphologically similar karyotypes. AB - The Japanese rose bitterling, Rhodeus ocellatus kurumeus, and the oily bitterling, Tanakia limbata, were cytogenetically studied by silver (Ag)- and chromomycin A3 (CMA3)-staining, by C-banding and by mapping of the 18S ribosomal genes and of the (TTAGGG)n telomeric sequence. These two representative species of related genera of the subfamily Acheilognathinae show very similar chromosome complements. Nevertheless, significant differences in the chromosomal distribution of nucleolus organizer regions (NORs) and interstitial telomeric sequences were observed. Whereas R. ocellatus kurumeus shows a single NOR-bearing chromosome pair, T. limbata is characterized by a higher number of variable NORs. Multiple telomeric sequence sites were found at the pericentromeric regions of several chromosomes in the rose bitterling. No telomeric sequence sites were detected near centromeres, but they were found to be scattered along the NORs in the oily bitterling. Two karyoevolutive trends might have been identified in the subfamily. PMID- 12903753 TI - Isobutyramide activates transcription of human fetal gamma- and murine embryonic epsilon(y)-globin genes. AB - OBJECTIVE: To examine the effect of isobutyramide synthesized in our laboratory on human and murine globin gene expression and to test cell toxicity of the drug. METHODS: MEL cells were transfected with the recombinant construct muLCRAgammapsibetadeltabeta and the stable transformants were cultured in the medium with different concentrations of isobutyramide. The experimental mice and rabbit were injected with different doses of isobutyramide. The globin mRNAs were analyzed by RNase protection assay. The hematological toxicity and electrolyte toxicity of the drug were tested. RESULTS: An inducible and dose-dependent expression of the human gamma-, beta- and mouse alpha-globin gene was observed in the transfected MEL cells. The induction of the human gamma-globin gene is significant stronger than that of the beta-globin gene. With 2.5 approximately 5 mmol/L isobutyramide, the induction of the human gamma-globin gene is even more effective than that of mouse alpha-globin gene. After a 15-day injection under the doses of 500 approximately 900 mg x kg(-1) x d(-1), the level of the mouse embryonic epsilon(y)-globin mRNA could be significantly induced up to 3 approximately 4 fold of that of uninjected controls. The changes of hemoglobin(Hb), RBC, hematocrit(HCT), WBC, derived from mice injected with different doses of isobutyramide at the interval of 24 hours for 2 approximately 4 weeks, were generally within the normal range. In rabbits injected with isobutyramide in the same regiment for 2 weeks, the concentration of blood K+, Na+, Cl- and CO2 were all within normal range and serum ionic osmotic pressure remained stable as well. CONCLUSION: Our results suggested that isobutyramide is a weak inducer of cell differentiation, but it can selectively activate transcription of human gamma-globin gene at a certain degree, and it can act on early stages of erythroid progenitor differentiation in adult mice and activate transcription of embryonic epsilon(y)-globin gene and have no hematological toxicity. Our results have further proved the potential value of isobutyramide in treatment of beta-thalassemia and sickle cell disease. PMID- 12903754 TI - The tree shrew apolipoprotein C-I cDNA: sequence and its expression. AB - A rabbit anti-serum to tree shrew apolipoprotein C-I (apo C-I) was used to screen an expression cDNA library constructed by us from tree shrew (TS) liver tissue. Two apo C-I cDNA clones were obtained. The longer one consists of 380 nucleotides, including 21 bp and 95 bp at the 5' and 3' end of the non-translated regions respectively, and a 264-bp fragment in an open reading frame encoding 88 amino acids prepropeptide which contains 26 amino acids of signal peptide and a mature protein (62 amino acids). Comparing the amino-acid sequence deduced from this cDNA with those of the published mammalian apo C-Is reveals that it shared some structural similarity with rat, mouse and dog apo C-I, but it had 5 more amino acids than that of human and baboon. The expression of apo C-I mRNA in 8 different tissues were also assayed with Northern blot. The results demonstrated that liver had the highest expression, intestine had much less expression and no expression in other tissues, which is much different from human and other species. This study has laid down a good foundation for further studying on the function and the stucture of tree shrew apo C-I gene. PMID- 12903755 TI - Expression and deletion analysis of EcoRII endonuclease and methylase gene. AB - OBJECTIVE: To clone complete EcoRII restriction endonuclease gene (ecoRIIR) and methyltransferase gene (ecoRIIM) in one vector and to analyze the coordinating expression of this whole R-M system. METHODS: Unidirectional deletion subclones were constructed with ExoIII. ecoRIIR/ M genes were preliminarily located in the cloned fragment according to the enzyme activities of subclones. Exact deletion sites were determined by sequencing, and transcriptional start sites were determined by S1 mapping. RESULTS: The DNA fragment which was cloned into pBluescript SK + contained intact ecoRIIR gene and ecoRIIM gene, and two transcriptional start sites of ecoRIIR gene were determined. 132bp to 458bp from 3'end of ecoRIIR gene are indispensable to enzyme activities and deletion of 202bp from 3' end of ecoRIIM gene made enzyme lose the capability in DNA protection to resist specific cut with EcoRII endonuclease (EcoRII. R). Deletion of the coding and flanking sequences of one gene did not affect the expression of the other gene, and the recombinants only containing ecoRIIR gene appeared to be lethal to dcm+ host. CONCLUSION: ecoRIIM gene linking closely to ecoRIIR gene is very important for the existence of the R-M system in process of evolution, but the key to control EcoRII R-M order may not exist in transcriptional level . PMID- 12903756 TI - The effect of ischemic re-perfusion injury plus particle infusion embolism on the apoptosis of rats with pancreatic cancer. AB - OBJECTIVES: In an attempt to develop new method of treating the end- or mid-stage pancreatic cancer, we examined the effect of ischemic re-perfusion injury plus particle embolism on the pathology and cell apoptosis of pancreatic cancer in Sprague Dawely rats. METHODS: 9 mg dimethylbeneanthracine (DMBA) were implanted directly into the parenchyma of pancreatic tail of Sprague-Dawely rats. After establishment of tumor, the inferior splenic artery, a main supplying vessel to pancreatic tail was subjected to blockade and re-opening for 30 min separately, then embolism particles were infused via the artery. Afterwards, artery was ligated. Pathological changes and cell apoptosis indicators (AI) of pancreatic cancer were observed by light microscopy and ISEL respectively 14 days after the operation. RESULTS: The prevalence of pancreatic cancer among DMBA-implanted rats evaluated 3 months to 4 months after implantation was 59%. The volumes of the tumor in positive control group (B), pancreatic ischemic group (C), pancreatic ischemic re-perfusion injury group (D) were significantly larger than pancreatic ischemic re-perfusion injury plus particle thrombus group (E) ( P < 0.01). The volumes of the tumor in groups D, E were significantly smaller than that in group C (P < 0.01). There was a significant difference in tumor size between group B and group C ( P < 0. 01), but the difference was not significant between group D and group E ( P > 0.05). There was a significant infiltration of tumor tissue in group B rats, but strong inflammatory reaction was not noted. In groups C, D, E, a localized tumor growth was observed; infiltration of inflammatory cells and proliferation of fibroblasts and connective fiber were obvious, and some of these fibers grew into cancer nests and separate the tumor. The above findings were most conspicuous in group E. There was a significant difference in AI between group E (13.7 +/- 1.5) and other groups ( P < 0.01), with the difference being also significant between group C (4.3 +/- 2.4), D (8.5 +/- 1.1) and group B (1.2 +/- 0.8) (P < 0.01), and between group C and group D (P < 0.01) or between group D and group E ( P < 0.01). In the samples of group A, the apoptotic cells were not found. CONCLUSIONS: Pancreatic ischemic re-perfusion injury plus particle thrombus can cause significant infiltration of inflammatory cells in tumor tissues thereby limiting its growth, and inducing cell apoptosis of pancreatic cancer. This effect is superior to either pancreatic ischemia alone or pancreatic ischemia plus re-perfusion injury. PMID- 12903757 TI - Gut barrier function damage following multiple firearm injuries in a porcine model. AB - OBJECTIVE: To study the characteristics and pathogenesis of gut barrier damage following multiple firearm injuries in a porcine model. METHODS: Twenty-four small pigs were divided into 4 groups: control group (n = 6, group C), group H (n = 6, gunshot-induced tangential fracture of parietal bone), group L (n = 6, gunshot-induced comminuted fracture of bilateral femora) and group M (n = 6, combined group H + L). Gastric intramucosal pH (pHi), plasma endotoxin levels in portal vein, and plasma D-lactate levels were measured and blood samples were cultured at different intervals after trauma. The animals were sacrificed at 72 h following trauma and intestinal tissues were harvested for pathological examination and diamine oxidase (DAO) activity measurement. RESULTS: In group M at 72 h, pHi was significantly lower than that of group H and L ( P < 0.01), and plasma endotoxin level was significantly higher than that of group H (P < 0.01) and group L (P < 0.05). Simultaneously, in group M, D-lactate level was markedly higher than that of group H ( P < 0.01), and incidence of positive blood culture was much higher than that of group H and L ( P < 0.05). Necrosis and exfoliation were revealed at ileum villus top in all trauma groups, especially in group M, in which ileum DAO activity declined most significantly as well. CONCLUSION: Multiple trauma is prone to cause gastrointestinal ischemia even without hemorrhagic shock. The damage of gut barrier in multiple trauma appears to be more severe than that in one-site trauma, thereby promoting gut-derived endotoxemia and bacterial translocation and contributing to the development of endogenous infection. PMID- 12903758 TI - Modulation of nitric oxide on lymphokine-activated killer cells in patients with bladder cancer. PMID- 12903760 TI - Preservation of the hypothalamic structures in the total resection of craniopharyngioma. AB - OBJECTIVE: To explore an effective method of surgical management of craniopharyngioma. SUBJECTS AND METHODS: Fifty patients with craniopharyngioma had total and subtotal tumor ectomy. There were 29 males and 21 females, ranging in age from 15 to 56 years (mean 34.1 years). MR imaging showed that the tumors were located in the superior sellar region in 24 cases, in superior sella region and extended into the third ventricular floor in 19 cases, into parasella in 3 cases and down to intrasella in 4 cases. Complete cystic tumors were found in 5 cases, while the partial cystic tumor in 24 cases and complete solid tumors in 21 cases. Pterional approach was used in 48 patients and subfrontal approach in 2 patients. Great attention was paid to the preservation of the perforating arteries from the carotid, posterior and anterior communicating and anterior choroidal arteries to the hypothalamic structures. The clinical outcome was evaluated according to the GOS scale. RESULTS: Of the 50 patients surgically treated, 47 patients obtained total ectomy of the tumor and 3 patients with the secondary surgery had subtotal ectomy of the tumor. The pituitary stalk was preserved in 29 (58%) patients, severed in 14 patients and unidentified in 7 patients. Forty-six patients regained a normal life; one patient needed assist in life. Of the 3 deaths, one patient died of diabetes insipidus, one of inhalation asphyxia, and another one of water and sodium disorders. CONCLUSION: Avoidance of the injury to the neural structures in the third ventricular floor and preservation of the perforating arteries to hypothalamus are the key to achieve good surgical results in treating craniopharyngioma. PMID- 12903759 TI - Surgical treatment of malignant esophageal tumors in PUMC Hospital. AB - To study how to prolong the postoperative survival time of the patients with malignant esophageal tumors. The clinical data of 1098 patients with malignant esophageal tumors from 1961 to 1992 were retrospectively analyzed. The deletion of fragile histamine triplet (FHIT) gene (a tumor suppressor gene) in 30 fresh esophageal samples obtained in 1996 was detected with PCR and RT-PCR method. The resectability was raised gradually and the operative morbidity and mortality decreased year by year, but there was no significant improvement on the postoperative 5-year survival rate. Delayed diagnosis and irradical resection influenced the long-term survival. The deletion of cDNA of FHIT gene was 64. 2% in esophageal cancer and 20% in the resected margin of the cancer. We believe that high-grade atypical hyperplasia in esophageal epithelium and deletion of FHIT gene in esophageal cancer and its resected margin are pathological and molecular markers for early diagnosis of esophageal cancer respectively, and the latter may be one of the molecular markers for the resection. Early diagnosis and treatment, radical, resection, and postoperative nutritional support are very important for the improvement of the postoperative survival time of the patients. PMID- 12903761 TI - Potentiation of docetaxel antitumor activity by batimastat against mouse forestomach carcinoma. AB - OBJECTIVE: To improve the treatment of tumors, we studied the combined effects of docetaxel and batimastat (BB-94) on mouse forestomach carcinoma (MFC), and compared them with doxorubicin. METHODS AND RESULTS: In vitro, growth curve analysis, MTT assay and clonogenic assay used to determine the cytotoxic effect of docetaxel or/and BB-94 on MFC cell showed that docetaxel but not BB-94 had a significant cytotoxicity, and the effect of docetaxel wasn't enhanced by BB-94. In early stage MFC tumor model, obvious antitumor effect of docetaxel or doxorubicin given i.v. at maximum tolerated dose (MTD, docetaxel: 20 mg/kg; doxorubicin: 6 mg/kg) every 4 days for 3 injections (q4d x 3), even that of BB-94 (30 mg/kg i. p. qd x 20) was observed. Tumor growth inhibition was greater for docetaxel-batimastat (96.0%) than for doxorubicin-batimastat (88.0%), docetaxel (89.0%), doxorubicin (68.0%) and BB-94 (33.0%), and the effect of docetaxel could be potentiated by BB-94. Docetaxel also showed activity against advanced stage MFC tumor in dose-dependent manner, and was more effective at MTD than doxorubicin with 4/5 regressions, 46.5 days tumor growth delay and 2.8log(10) tumor cell kill. CONCLUSION: Our results suggest that in the MFC model with dose and schedule used, docetaxel is an effective cytotoxic new drug against MFC tumor and BB-94 enchances the antitumor activity of docetaxel. PMID- 12903762 TI - Thoracic spine fractures. AB - OBJECTIVE: To investigate the unique characteristics and treatment of thoracic spine fractures. METHODS: Seventy-seven patients with thoracic spine fractures were retrospectively reviewed. Of these, there were 37 compression fractures, 34 fracture-dislocations, 3 burst fractures and 3 burst-dislocations. Twenty-six patients had a complete lesion of the spinal cord, 14 sustained a neurologically incomplete injury, and 37 were neurologically intact. Fifty-three patients were treated nonoperatively and 24 treated operatively. RESULTS: All patients were followed up for 2 approximately 15 years. None of the 26 patients with a complete lesion recovered any significant function. Of 37 neurologically intact patients, 13 had local pain although all of them remained normal function. Two of 14 patients with incomplete paraplegia returned to normal, 7 recovered some function and 5 did not recovered. CONCLUSIONS: Because of the unique anatomy and biomechanics of the thoracic spine, the classification commonly applied to thoracolumbar fractures is not suitable for thoracic fractures. Fusion and instrumentation are indicated when the fractures are unstable, while patients with incomplete lesion of the spinal cord may be the candidates for supplemented decompression. PMID- 12903763 TI - Urotensin II receptor in the rat airway smooth muscle and its effect on the rat airway smooth muscle cells proliferation. AB - OBJECTIVE: To investigate the characteristics of urotensin II (U-II) receptor in the rat airway smooth muscle and the effect and signal transduction pathway of U II on the proliferation of airway smooth muscle cells. METHODS: Using 125I-UII binding assay to measure the Bmax and Kd of U-II receptor. Using the 3H-TdR incorporation to determine the effect of U-II on the proliferation of airway smooth muscle cells and its signal transduction pathway. Using Fura-2/AM to measure the effect of U-1I on the cytosolic free calcium concentration. RESULTS: 1. 125I-UII binding increased with the time and reached saturation at 45 min. The B(max0 was (11.36 +/- 0.37)fmol/mg pr and Kd was (4.46 +/- 0.61) nmol/L. 2. U-II increased 3H-TdR incorporation of the airway smooth muscle cells in a dose dependent manner. 3. H7, PD98059 and nicardipine, inhibitors of PKC, MAPK, calcium channel, respectively, significantly inhibited U-II-stimulated 3H-TdR incorporation of airway smooth muscle cells. W7, inhibitor of CaM-PK, had no effect. 4. Cyclosporin A, inhibitor of CaN, inhibited 3H-TdR incorporation of the airway smooth muscle cells induced by U-II in a dose-dependent manner. 5. U-II promoted cytosolic free calcium concentration increase by 18%. CONCLUSIONS: 1. There was U-II receptor in the rat airway smooth muscle. 2. The effect of U-II stimulated 3H-TdR incorporation of airway smooth muscle cells was mediated by such signal transduction pathway as Ca2+, PKC, MAPK and CaN, etc. PMID- 12903764 TI - Evaluation of the diagnostic criteria for the localization of acquired arteriovenous fistulas by color Doppler flow imaging. AB - OBJECTIVE: To evaluate the diagnostic criteria for the localization of acquired arteriovenous fistulas (AVFs) by color Doppler flow imaging (CDFI) based on the features of their hemodynamic changes. METHODS: The shape and hemodynamic changes of involved vessels which could be helpful to localize the sites of fistulas were studied according to the observation of 10 cases of acquired AVFs. RESULTS: The sites of the fistulas could be shown by two-dimensional ultrasonography and color flow imaging in 40% and 80% cases, respectively. In all cases, turbulent high velocity flow was present at the sites of the fistulas, low resistant flow was present in the arteries proximal to the fistulas, and artery-like flow was detected in the veins. CONCLUSION: CDFI was accurate for the localization of acquired AVFs, which were mainly localized by their hemodynamic changes shown by pulse Doppler ultrasound. PMID- 12903765 TI - The management of epithelial ingrowth after laser in situ keratomileusis. AB - OBJECTIVE: . To investigate the treatment of corneal epithelial ingrowth with flap melt after LASIK. PATIENTS AND METHOD: Thirteen eyes of 13 patients with corneal epithelial ingrowth and flap melt after LASIK were treated based on the thickness and the extent of melting of the flap. Nine eyes were treated by lifting the flap and scraping the epithelium in the interface. Three eyes underwent allogenic keratomileusis. The flap was removed in one eye. RESULTS: Recurrence occurred in 2 eyes treated by lifting the flap. The flaps, however, healed normally after retreatment. The grafts healed normally in the allogenic keratomileusis group, except for one eye in which the flap was lost. The corneal epithelium healed over the stromal bed after the flap was removed. Corneal haze of grade 2 was present at 6 months. The uncorrected visual acuity was between 0.3 and 1.0, and the best-corrected visual acuity lost 1 to 3 lines. CONCLUSIONS: Corneal epithelial ingrowth with flap melt is a severe complication after LASIK. However, with proper treatment, vision can be salvaged in most cases. PMID- 12903766 TI - Relation between alkaline phosphatase in gingigval crevicular fluid of implant teeth and the curing result. AB - OBJECTIVE: To discover the relation between alkaline phosphatase (ALP) in gingival crevicular fluid (GCF) of implant teeth and the curing results. METHODS: We measured the ALP level in GCF among 56 cases of implant teeth which included 2 failed cases, 5 cases with bad oral hygiene and gingivitis, and compared it with that in the normal group composed of 10 persons. RESULTS: The ALP levels in normal group and success implant group showed no difference. The ALP levels in normal group and success with gingivitis group showed obvious difference. The ALP levels of the 2 failed cases are the highest of all. CONCLUSIONS: The ALP level in GCF is an important index in evaluating the curing result of the implant teeth. PMID- 12903767 TI - The past and future of clinical nutritional support in China. PMID- 12903768 TI - Effect of Ginkgo biloba extract on brain edema after subarachnoid hemorrhage in rats. PMID- 12903769 TI - Effect of VL and VH consensus sequence-specific primers on the binding and expression of a mini-molecule antibody directed towards human gastric cancer. AB - OBJECTIVE: To construct ScFv and Fab from murine anti-gastric cancer monoclonal antibody (mAb)3H11. METHODS: At first, 3H11 ScFv and Fab were constructed with V genes PCR amplified by degenerate primers for FR1. The bacterial expressed 3H11 Ab fragments showed no antigen binding activity. Then, phage antibody library and random mutated library were constructed from 3H11 hybridoma cells and panning selection was perfomed. Again the identification of positive clone was failed. Finally the N-terminal sequences of V regions were resumed to 3H11 original sequences by site-directed mutagenesis via PCR. RESULTS: Binding activity to gastric cancer cells was detected only from N-terminal sequence corrected 3H11 ScFv and Fab,though the expression of the Ab fragments was not affected. Correction of either VL or VH N-terminal sequences could partially resume the antigen binding activity. CONCLUSION: Sequence changes of V region N terminal introduced by PCR may seriously affect antigen binding without affecting the expression of antibody. PMID- 12903770 TI - Inhibitory effect of fluvastatin on aortic intimal thickening in normocholesterolemic rabbits. AB - OBJECTIVE: The anti-atherosclerotic effect of fluvastatin at does insufficient to lower serum cholesterol on the catheter-induced intimal thickering and possible mechanism were investigated in abdominal aorta of rabbits. METHODS: Fifty-six rabbits were randomly divided into eight groups (n = 7, each). Fluvastatin was given mixed with food at daily dose of 8 mg/kg starting 5 days before catheterization. Light microscope, immunohistochemistry, transmission electron microscope and RT-PCR assay were applied to assess vascular smooth muscle cell (VSMC) proliferation and apoptosis, as well as oncogene expression in vascular wall. RESULTS: At day 10 and day 15 after catheter induced denudation intima/media (I/M) thickness ratio was obviously higher, and also the percentage of PCNA-positive cells and TUNEL-positive cells in media was significantly higher compared with controls. The intimal hyperplasia was mostly composed of alpha-SM actin-positive cells. In rabbits given fluvastatin I/M ratio and the percentage of these positive cells significantly decreased compared with those without fluvastatin. The overexpression of proto-oncogene H-ras mRNA and decreased expression of anti-oncogene p53 mRNA were found after vascular injury, whereas fluvastatin significantly reduced H-ras mRNA and increased p53 mRNA expression. CONCLUSION: Proliferation of VSMC in the media and the migration to the intima can be inhibited, and apoptosis of VSMC be induced by short-term use of fluvastatin after balloon catheter denudation, independent of serum lipid change. The underlying mechanism is presumably associated with the influence of fluvastatin on oncogene expression in the injured vascular wall. PMID- 12903772 TI - Direct coronary stenting in acute myocardial infarction. PMID- 12903773 TI - Effect of coxsackievirus b3 on ion channel currents in rat ventricular myocytes. AB - OBJECTIVE: To investigate the effects of coxsackievirus B3(CVB3) on ion channel currents in rat ventricular myocytes. METHODS: Rat hearts were isolated with collagenase to acquire single ventricular myocytes, L-type voltage-dependent calcium channel (VDCC) current (I(Ca)), Na+ current (I(Na)), outward potassium current (I(out)), inwardly rectifying potassium current(I(KI)) were recorded using whole cell patch clamp techniques. RESULTS: CVB3 infection increased I(Ca) and I(out), while decreased I(KI); but it had no obvious effect on I(Na). CONCLUSION: The effects of CVB3 an I(Ca), I(out), I(KI) may be one of the mechanisms of myocytes damage and the occurrence of abnormal electroactivities induced by CVB3 infection. PMID- 12903774 TI - Dynamic changes of serum vascular endothelial growth factor levels in a rat myocardial infarction model. AB - OBJECTIVE: To investigate the dynamic changes of serum vascular endothelial growth factor (VEGF) levels in a rat model of acute myocardial infarction. MATERIALS AND METHODS: Eighty-eight adult male Sprague-Dawley rats weighing approximately 270 g we used in this study. Eighty rats were subjected to left coronary artery ligation, with 8 rats for each different duration of infarct. Eight sham-operated animals in which the left coronary artery was surgically exposed without ligation were used as controls. Blood samples were drawn from the right atrium before (sham animals) and 1, 3, 6, 12, 24 h and 2, 3, 5, 7, 14 d after myocardial infarction. The concentrations of serum VEGF were measured by a sensitive enzyme-linked imosorbent assay with a rabbit polyclonal antibody specific for VEGF. RESULTS: In the 8 control animals, the mean concentration of serum VEGF was 66.99 +/- 17.83 pg/ml. Six hours after myocardial infarction, the level of serum VEGF significantly increased to 125.68 +/- 28.07 pg/ml (P < 0.01 vs x sham controls), and reached a peak (240.61 +/- 70.63 pg/ml. P < 0.01 vs x sham animals) at 24 h after ligation and then decreased gradually over the remaining 2 weeks. However, the level remained significantly elevated for 14 d (107.64 +/- 30.13 pg/ml, P < 0.01 vs x sham controls). CONCLUSION: The present study shows that the levels of serum VEGF are markedly increased until 14 d in the rat model of acute myocardial infarction. The increased serum VEGF level may play an important role in the angiogenesis associated with myocardial infarction. PMID- 12903771 TI - Anti-human platelet tetraspanin (CD9) monoclonal antibodies induce platelet integrin alphaIIbbeta3 activation in a Fc receptor-independent fashion. AB - OBJECTIVE: This study characterized the activation of platelet integrin alphaIIbbeta3 induced by two anti-human platelet tetraspanin monoclonal antibodies (mAbs), HI117 and SJ9A4. METHODS: Using 125I-labeled human fibrinogen(Fg), specific Fg binding to human platelets induced by HI117 and SJ9A4 was measured as indication of activation of platelet integrin alphaIIbbeta3 by the two mAbs. RESULTS: HI117 and SJ9A4 (10 microg/ml and 20 microg/ml) induced evident specific Fg binding to human platelets, suggesting that the two mAbs evoked activation of platelet integrin alphaIIbbeta3. Further study indicated that HI117 and SJ9A4 induced integrin alphaIIbbeta3 activation independent of platelet Fc-receptors, and that HI117 and SJ9A4-induced integrin alphaIIbbeta3 activation was inhibited by sphingosing, aspirin, apyrase, and/or PGI2. CONCLUSION: The anti-platelet tetraspanin (CD9) mAbs, HI117 and SJ9A4, can induce platelet integrin alphaIIbbeta3 activation independent of Fc-receptor. Three signaling pathways, i.e. thromboxane, secreted ADP, and cAMP pathways may be involved in the process, with protein kinase C activation presumably being the common step of the three pathways. PMID- 12903775 TI - Arginine vasopressin gene expression in supraoptic nucleus and paraventricular nucleus of hypothalamous following cerebral ischemia and reperfusion. AB - BACKGROUND: Our previous studies indicated that the increased arginine vasopressin (AVP) in ischemic brain regions of gerbils could exacerbate the ischemic brain edema. This experiments is further clarify the relation between AVP and cerebral ischemia at the molecular level. METHODS: The contents of AVP, AVP mRNA, AVP immunoreactive (ir) neurons in supraoptic nucleus(SON) and paraventricular nucleus (PVN) after cerebral ischemia and reperfusion were respectively determined by radioimmunoassay (RIA), immunocytochemistry (II C), situ hybridization and computed image pattern analysis. RESULTS: The contents of AVP in SON, PVN were increased, and the AVP ir positive neurons in SON and PVN were also significantly increased as compared with the controls after ischemia and reperfusion. And there were very light staining of AVP ir positive neurons in the other brain areas such as suprachiasmatic nucleus (SC) and periventricular hypothalamic nucleus (PE), but these have no significant changes as compared with the controls. During different periods of cerebral ischemia (30 approximately 120 min) and reperfusion (30 min), AVP mRNA expression in SON and PVN were more markedly increased than the controls. CONCLUSIONS: The transcription of AVP gene elevated, then promoting synthesis and release of AVP in SON, PVN. Under the specific condition of cerebral ischemia and reperfusion, the activity and contents of central AVP increased abnormally is one of the important factors which causes ischemia brain damage. PMID- 12903776 TI - The bi-directional regulation of filamin on the ATPase activity of smooth muscle myosin. AB - OBJECTIVE: The aim of this study is to investigate the functional relationship between filamin, a known actin binding protein, and myosin and the effects of filamin on the interaction between myosin and actin. METHODS: Ultra centrifugation method was used to investigate the binding of filamin to both phosphorylated and unphosphorylated myosins. Mg-ATPase activities of both phosphorylated and unphosphorylated myosins in the presence and absence of actin were measured to observe the effects resulted from filamin-actin and filamin myosin interactions. RESULTS: It was found that filamin is also a myosin binding protein. Filamin inhibited the actin activated Mg-ATPase activity of phosphorylated myosin and stimulated Mg-ATPase of phosphorylated myosin in the absence of actin; in addition, filamin stimulated Mg-ATPase activity of unphosphorylated myosin in both the presence or absence of actin. CONCLUSION: The result suggest that the effects of filamin on the myosin Mg-ATPase activities are bi-directional, i.e., stimulatory via binding to myosin and inhibitory via binding to actin. PMID- 12903777 TI - Effect of kanglemycin C on lymphokin a production and gene expression of mouse splenocyte and macrophage. AB - AIM: To study the effect of kanglemycin C (KC) on production and gene transcription of lymphokins. METHODS: Cell proliferation and lymphokin activities were quantified with MTT colorimetry and ELISA, and gene transcriptions of lymphokins semi-quantified with RT-PCR. RESULTS: Suppression of KC on proliferation of enriched T-and B-cell respectively mediated by Con A and LPS was declined by addition of exogenous IL-1, IL-2, and IL-6. KC 80 nmol/L markedly inhibited IL-2 and IL-6 production and mRNA transcription of incubated mouse splenocytes induced by Con A. Additionally, KC had some suppression on IL-1beta and IL-6 productions of peritoneal macrophage stimulated by LPS (5microg/mL), whereas cyclosporine (CS) had not. CONCLUSION: Immunosuppression of KC came true partially through the decrease of IL-1beta, -2 and -6 productions, especially of IL-2. However, CS's immunosuppression was mainly through the decrease of IL-2 production. PMID- 12903778 TI - Molecular cloning of hTRT catalytic domain from HeLa cells and its expression in E. coli and purification. AB - OBJECTIVE: To investigate the expression of telomerase gene hTRT mRNA in HeLa cells and to obtain hTRT protein for further study. METHODS: The gene for encoding hTRT catalytic domain was cloned based on RT-PCR amplification from HeLa cells and sequenced. The cloned hTRTcDNA was in-frame inserted into His-tag fusion expression vector pEK318. The His-tag hTRT fusion protein were purified by Ni-NTA chromatography and stained by western blotting. RESULTS: An approximately 620bp fragment was generated and cloned into pBluescript SK + between SaII and BamHI sites. DNA sequencing showed the isolated fragment was consistent to those reported. SDS-PAGE present that a 17kDa protein was expressed stably in E. coli JM109 harboring pEKTRT344 containing 6 x His-tag and hTRT 150aa, and the expression level of the protein was about 26% of the total bacterial proteins, while the expression of pEKTRT containing 6 x His-tag and hTRT 243aa was only detectable as 27 kDa band in western blotting. Both of fusion proteins were purified by Ni-NTA chromatography and showed single band( > 95% purifity) in Coomassie Brilliant staininng. Western-blotting confirmed that two proteins could be recognized by the Ni-NTA AP conjugate. CONCLUSIONS: The hTRT catalytic domain was highly conserved. The expressed hTRT protein contained recognizable His-tag, telomerase-specific and strong antigenic epitops, which may be convenient for further investigation. PMID- 12903779 TI - Expression of T cell receptor V(alpha) gene families in intrathyroidal T cells of Chinese patients with Graves' disease. AB - OBJECTIVE: Patients with Graves' disease (GD) have marked lymphocytic infiltration in their thyroid glands. We examined the gene for the variable regions of the alpha-chain of the Chinese T-cell receptor (V(alpha) gene) in intrathyroidal T cells to determine the role of T cells in the pathogenesis of GD and offer potential for the development of immunotherapeutic remedies for GD. METHODS: We used the reverse transcription and polymerase chain reaction (RT-PCR) to amplify complementary DNA(cDNA) for the 18 known families of the V(alpha) gene in intrathyroidal T cells from 5 patients with Graves' disease. The findings were compared with the results of peripheral blood T cells in the same patients as well as those in normal subjects. RESULTS: We found that marked restriction in the expression of T cell receptor V(alpha) genes by T cells from the thyroid tissue of Chinese patients with GD(P < 0.001). An average of only 4.6 +/- 1.52 of the 18 V(alpha) genes were expressed in such samples, as compared with 10.4 +/- 2.30V(alpha) genes expressed in peripheral blood T cells from the same patients. The pattern of expressed V(alpha) genes differed from patient to patient with no clear predominance. CONCLUSIONS: Expression of intrathyroidal T cell receptor V(alpha) genes in GD is highly restricted suggesting the primacy of T cells in causing the disorders. PMID- 12903780 TI - Closed reduction and percutaneous K-wires fixation of displaced supracondylar humerus fractures in children. AB - OBJECTIVE: To observe the effects of closed reduction and percutaneous K-wires fixation of displaced supracondylar humerus fracture in children. METHODS: Retrospective review of fourteen patients who sustained displaced supracondylar fracture of distal humerus treated by closed reduction and percutaneous K-wires fixation. RESULTS: All patients' K-wires were removed at 4 weeks post-operation. Their elbow function regained at weeks. The average period of followed up was 10 month (varies from 6 to 18 month), all fractures healed without any permanent complications. Two transient nerves palsy, ulnar and radial nerve each, recovered 12 weeks and 16 weeks post-operation respectively. CONCLUSION: Closed reduction and percutaneous K-wires fixation is a safe and efficient treatment for humerus surpracondylar fracture in children. PMID- 12903781 TI - The operative treatment of spinal fracture-dislocation without neurologic deficits. AB - OBJECTIVE: To evaluate the results of operative treatment of spinal fracture dislocation without neurologic deficits. METHODS: Eighteen patients with spinal fracture-dislocation were neurologically intact at the time of injury, and all were treated operatively. The fracture sites were: 8 cases in cervical spine, 3 cases in thoracic spine, and 7 cases in lumbar spine. Eight patients with cervical injuries had variant degrees of forward slide and kyphotic deformity. Of the 10 thoracic and lumbar fractures, one had lateral dislocation, 4 cases with kyphotic deformities, 5 cases with spinal canal compromise averaged 50% (ranging from 40% to 70%). RESULTS: The average period of follow-up was 4.4 years with a range of 11 months to 13 years. All the patients returned to full-time work. No patient developed neurologic deterioration. Kyphotic deformity was corrected in the 4 cases, and no progressive kyphosis was noted. There was no operation related complication. The averaged post-operative hospitalization time was 13 days. CONCLUSION: Despite the rare incidence of spinal fracture-dislocation without neurologic deficits, we suggested that kind of fracture be considered unstable fracture because of its potential risk of delayed neurologic deterioration and kyphotic deformity, and be treated operatively to restore the sagittal alignment and the stability of the spine. PMID- 12903782 TI - Application of ambulatory blood pressure monitoring in essential hypertension with target-organ damage. PMID- 12903783 TI - Final report about Stage II Collaboration Project on Leprosy Rehabilitation. AB - OBJECTIVE: To undertake the pilot experiments of prevention of disability (POD) in 14 different geographical areas to serve as examples for future development of rehabilitation work in China and in Asia. METHODS: According to the principles and national criterion, 27 000 people affected by leprosy were selected and assessed using disability record forms at beginning and followed up regularly for observing changes of different indicators. RESULTS: A total of 197 neuritis cases were detected and treated with prednisolone out of 1 407 new or active cases. Self-care training of eyes, hands and feet were conducted for 10 500 disabled people affected by leprosy. Comprehensive therapy was given to 1 804 cases having complicated ulcers of which 1 055 cases have got their ulcers healed. Out of 706 prostheses, 613 were given to patients with satisfactory results. Surgical treatment was given to 269 cases and 251 have shown good progress. CONCLUSION: Most of patients have got benefit from the project in function or appearance which is very helpful for their going back to the society and agreed by foreign experts during the final evaluation. The experiences from the project can be implemented in the whole country. PMID- 12903784 TI - Study on modified cold storage method of rat livers with self-made hyd solution. AB - OBJECTIVE: To investigate the cold preservation effect of rat livers by modified storage method with self-made HYD solution. METHODS: The modified method was that the vascular bed of rat livers was expanded with an additional 20 to 40ml self made HYD solution/100g liver. After removing the liver, the extra HYD solution expressed as % liver weight was entrapped via portal infusion by tying off the supra-and infra hepatic inferior vena cava. According to the amount of extra HYD solution, 40 rats were randomly divided into four groups including: control group with conventional storage method, 20% group, 30% group and 40% group. The preservation effect of modified storage method with that of conventional storage method by using isolated perfused rat liver model was compared. RESULTS: Bile production and all the indices of hepatic microcirculation including portal perfusion pressure, endothelin-1 in the effluent, trypan blue distribution time and histology in modified method groups were significantly superior to those in control group (P < 0.05). The liver enzymes in 30% group were markedly lower than those in control group (P < 0.05). The preservation effect of rat liver in 30% group was the best among the modified method groups. CONCLUSION: The modified cold stage method is effective and may have potential for clinical application for liver preservation. PMID- 12903785 TI - Properties of voltage-gated sodium channels in developing auditory neurons of the mouse in vitro. AB - OBJECTIVE: To investigate the properties of voltage-gated sodium (Na+) channels in developing auditory neurons during early postnatal stages in the mammalian central nervous system. METHODS: Using the whole-cell voltage-clamp technique, we have studied changes in the electrophysiological properties of Na+ channels in the principal neurons of the medial nucleus of the trapezoid body (MNTB). RESULTS: We found that MNTB neurons already express functional Na+ channels at postnatal day 1 (P1), and that channel density begins to increase at P5 when the neurons receive synaptic innervation and reach its maximum (approximately 3 fold) at P11 when functional hearing onsets. These changes were paralleled by an age dependent acceleration in both inactivation and recovery from inactivation. In contrast, there was very little alteration in the voltage-dependence of inactivation. CONCLUSION: These profound changes in the properties of voltage gated Na+ channels may increase the excitability of MNTB neurons and enhance their phase-locking fidelity and capacity during high-frequency synaptic transmission. PMID- 12903786 TI - Rate-dependent slow conduction velocity in the cavo-tricuspid isthmus and septum in patients with atrial flutter. AB - PURPOSE: To evaluate and compare the effects of heart rate on conduction velocity in the cavotricuspid isthmus (CTI) and septum in patients with and without typical atrial flutter (AF) using electro-anatomic mapping (EAM) of the right atrium (RA). METHODS: Ten patients (age 53+/-10 yrs, 7M/3F) with AF and 13 patients (age 51+/-11 yrs, 5M/8F) with atrioventricular nodal reentrant tachycardia (AVNRT) underwent conventional electrophysiological study, electro anatomic mapping and radiofrequency ablation. Using EAMs obtained during coronary sinus pacing at pacing cycle length (PCL) 600 ms, 400 ms, and 300 ms, we evaluated conduction velocities in the CTI and septum of RA in 10 patients with AF and compared EAMs to 13 patients with AVNRT to determine whether the conduction slowing required to maintain AFL was related to changes in volume alone or altered RA electrophysiology. RESULTS: Conduction velocities in CTI and septum were significantly slower at all PCL when AF was compared to AVNRT (*P<0.05). Additionally, in the AF group, septal conduction velocities were slower at PCL 600 ms and 400 ms, but not at 300 ms compared to CTI (*P<0.05). In AF, during PCL 300, conduction in CTI slowed significantly compared to PCL 600 and 400 ms such that there was no difference between CTI and septum at PCL 300. CONCLUSIONS: There is slower conduction in the septum compared to the CTI in all patients. However, in patients with AF, there is significant slowing of conduction in the CTI and septum as well as decremental rate-dependent slowing of conduction in the CTI. These findings indicate that in addition to RA enlargement, changes in atrial electrophysiology distinguish AF patients from patients with AVNRT. PMID- 12903787 TI - Expression of matrix metalloproteinase-7 involving in growth, invasion, metastasis and angiogenesis of gastric cancer. AB - OBJECTIVE: To investigate the role of matrix metalloproteinase-7 (MMP-7) expression in caricinogenesis and progression of gastric cancer. METHODS: We studied MMP-7 expression and microvessel density (MVD) in adjacent mucosa and primary foci of 113 cases of gastric cancer by streptavidin-biotin immunoperoxidase method using anti-MMP-7 and anti-CD34 antibodies. MMP-7 expression and mean MVD were compared with clinicopathological features of gastric cancer, with the relationship between MMP-7 expression and MVD concerned in gastric cancer. RESULTS: MMP-7 showed positive expression in adjacent mucosa of gastric cancer (29.20%, 33/113), less than that in gastric cancer (69.03%, 78/113). MMP-7 expression in primary foci of gastric cancer was positively correlated with tumor size, invasive depth, metastasis and TNM staging (P<0.05), but not with differentiation or growth pattern of gastric cancer (P>0.05). Positive correlation of mean MVD with tumor size, invasive depth, metastasis and TNM staging was found (P<0.05), despite no relationship between mean MVD and differentiation of gastric cancer (P>0.05). Mean MVD was dependent on MMP-7 expression in gastric cancer (P<0.05). CONCLUSION: Upregulated expression of MMP 7 played an important role in carcinogenesis and progression by participating in growth, invasion, metastasis and angiogenesis of gastric cancer. MMP-7 expression could be regarded as an effective and objective marker to reflect the biological behaviors of gastric cancer. PMID- 12903788 TI - The association of the expression of MTA1, nm23H1 with the invasion, metastasis of ovarian carcinoma. AB - OBJECTIVE: To understanding the molecular mechanisms in invasion and metastasis of the ovarian carcinoma, we investigate a novel candidate metastasis-associated gene (MTA1) and nm23H1 mRNA expression and mutation in ovarian carcinoma. METHODS: Twenty primary ovarian carcinoma specimens, 20 corresponding lymph nodes and 8 normal ovarian was examined for mRNA expression and mutation of MTA1 and nm23H1 genes by reverse-transcription polymerase chain reaction (RT-PCR) and RT PCR-SSCP analysis. The level of the expression was determined by the relative optic density (ROD) of the PCR products. RESULTS: The frequency of MAT1 overexpression was 100% (7/7) in primary ovarian carcinoma with metastasis but only 38.5% (5/13) in those without metastasis (P=0.0103). Overexpression of MAT1 was observed in 87.5% (6/7) of lymph nodes with metastasis but only 23% (3/13) of lymph nodes without metastasis (P=0.0118). In contrast with MAT1, low expression of nm23H1 mRNA was seen in 7 of 7 ovarian carcinoma with metastasis but only in 4 of 13 (30%) of those without metastasis (P=0.0043). Low nm23H1 expression was also seen in 7 of 7 lymph nodes with metastasis but only in 5 of 13 (38.5%) nonmetastatic lymph nodes (P=0.0102). The ROD ratio of MAT1 to nm23H1 increased with the development of metastasis. No mutation of MAT1 and nm23H1 genes was found by SSCP analysis. CONCLUSION: The mRNA expression of MTA1 and nm23H1 is positively and negatively correlated with lymph node metastasis, respectively. Expression abnormalities but not mutation of the two genes are frequent events related to lymph node metastasis of ovarian cancer. PMID- 12903789 TI - The biological research of clinical behavior of laryngeal cancer. PMID- 12903790 TI - Evaluation of surgery in simple ovarian cysts. AB - OBJECTIVE: To evaluate the surgery in simple ovarian cysts. METHODS: From Jan. 1994 to Dec. 1999, 221 women with simple ovarian cysts were admitted into our hospital. The diameter of cysts was < 5 cm in 76 cases, and was > or = 5 cm in 145 cases ultrasonically. One hundred and eighty-four patients underwent laparoscopy, and thirty-seven underwent laparotomy. RESULTS: Histological findings showed no malignancy in this series. Simple cysts, paraovarian cysts and corpus luteum were found histologically in 90.8% of cases with cysts < 5 cm, and 60% of those with cysts > 7 cm respectively, their difference was significant (chi2=37.13, P<0.001). The simple cysts, paraovarian cysts and corpus luteum cysts were found in 81.5% of postmenopausal women and 84.0% of premenopausal women. CONCLUSION: Patients with cysts > 7 cm are indicated for surgical procedures, while a period of followup is acceptable for patients with cysts < 7 cm, and surgery is advisable if the cyst is persistent during followup. Postmenopausal women with cysts should have operations. PMID- 12903791 TI - Bone mineral analysis of whole body in 292 normal subjects assessed by dual X-ray absorptiometry. AB - OBJECTIVE: To study the whole body bone mineral changes in normal subjects. METHODS: Bone mineral was measured by dual X-ray absorptiometry (DXA) in 292 normal subjects aged 10-79 years, including 152 females and 140 males. They were divided into age-groups by decade. RESULTS: In males, the ages with peak value of total bone mineral content (TBMC), total bone mineral density (TBMD), anteroposterior spine BMD (L2-4 BMD) were in the 30-39, 20-39, 20-29 age-groups respectively. In females, they were all in the 30-39 age-group. Peak values of TBMC, TBMD, L2-4 BMD were higher in males than those in females. Loss of bone mineral for females was more pronounced than that for males. Loss of L2-4 BMD was more pronounced than TBMD. CONCLUSION: This study provides the normal bone mineral data of whole body for males and females, and the characteristic of changes. PMID- 12903792 TI - Peritoneal dialysis after repair of congenital heart disease in children. AB - OBJECTIVE: To investigate the effect of peritoneal dialysis on fluid balance and outcome in children who receive cardiac operation. METHODS: From July to Dec. 2000, 12 (2.08%) patients of 576 consecutive children who underwent heart operation required peritoneal dialysis because of acute renal failure. The mean age of these 12 patients was (2.9+/-2.0) years (range, 5 months-7 years) and the mean body weight was (12+/-3) kg (range, 7.4-18.5 kg). RESULTS: The interval between the operation and the initiation of peritoneal dialysis was (21.2+/-11.4) hours (4.4-42 hours). The duration of peritoneal dialysis was (6.3+/-4.8) days (0.47-15 days). Mortality in these 12 patients was 25%. Fluid removed by peritoneal dialysis was (34.7+/-17.8) ml x kg(-1) x day(-1). Asymoptomatic hypokalemia, thrombocytopenia and hyperglycemia were frequent complications, which were easily managed. Hemodynamics and pulmonary function improved during the study period. CONCLUSION: The early initiation of peritoneal dialysis is an effective and safe method to treat acute renal failure after cardiac operation in children. PMID- 12903793 TI - The analysis of changes and influencing factors of early postthoracotomy pulmonary function. AB - OBJECTIVE: To investigate the changes and influencing factors of early postoperative pulmonary function of thoracotomy. METHODS: Pre- and early postoperative pulmonary function was studied in 64 consecutive cases with optimal thoracotomy. Pain assessment was done before pulmonary function test, and the chief complaints of patients were recorded after the procedure. The changing curves of pulmonary function were done and the differences associated with groups, surgical styles, pain assessment, epidural analgesia, chief complaint and preoperative conditions were analyzed. RESULTS: Pulmonary function was severely lowered to about 40% of the base line on the first day, and it was rehabilitated to about 60% of the base line on the eighth day. There was a greater gradient on the recovery curve on the 3rd and 4th days. Epidural analgesia was able to improve pain relaxation and pulmonary function in some degree. Single-factor analysis showed that postoperative pain, postoperative day and surgical style were the significant influencing factors for early postoperative pulmonary function. By multiple-factor analysis, preoperative pulmonary function, age and postoperative pain were the main factors, while surgical style had only weak effect on it. CONCLUSIONS: Early postoperative pulmonary function is severely impaired by thoracotomy. It rehabilitate gradually with time. Improvement of preoperative pulmonary function, reducing surgical procedure injuries, especially injury to respiratory muscle system, and enough postoperative pain relief are the most important means that would reduce pulmonary function impairment and consequently reduce postoperative pulmonary complications. PMID- 12903794 TI - Interventional or semi-interventional treatment for Budd-Chiari syndrome. AB - OBJECTIVE: Report the results of interventional or semi-interventional techniques for 173 patients with Budd-Chiari syndrome. METHOD: This group included 120 males and 53 females. The pathologic lesions composed of localized complete occlusion of inferior vena cava (IVC) (78), IVC stenosis (49), IVC membrane with a hole (37), membrane of hepatic vein (HV) (3), IVC thrombosis (4), IVC membrane with thrombosis (2) and IVC lesion with occlusion of HV (32). Treatment methods included that I: Percutaneous transinferior vena cava angioplasty (PTA) (76); II: IVC PTA with stent (59); III: Percutaneous transhepatic vein recanalization (3); IV: IVC thrombolysis through a catheter (4), V: Combined transcardiac and transfemoral venous membranotomy and balloon dilation (22); VI: V and stent (17); VII: Stenting during radical surgery (3); VIII: Additional operation after intervention (23). RESULTS: The immediate technique success rate for intervention was 90.1%, for the semi-intervention was 100%. The IVC pressure was reduced from 3 to 29 cmH2O. Complications occurred in 8 cases. The death rate was 2.9%. A follow-up study showed the recurrence rates were 14.5% in IVC PTA group, 1.7% in IVC PTA with stent, 18.2% in combined technique without stent and no recurrence was found in other groups. CONCLUSION: The PTA is the first choice for localized lesions. When elastic recoil occurs, immediate stenting is suggested. The semi interventional approach is advised for PTA failure and more complicated cases. For those with both IVC lesion and occlusion of HV, the additional operation is needed after IVC intervention. PMID- 12903795 TI - Experience on surgical management of rupture of abdominal aortic aneurysm. AB - OBJECTIVE: To describe our surgical experience on rupture of abdominal aortic aneurysm. METHODS: Two cases of ruptured aortic aneurysms with severe complication were analysed. Aorta reconstruction procedures were performed using bifurcated e-PTFE grafts during emergency operation. Diagnosis, preoperative resuscitation, emergency surgical intervention, and postoperative complications of these patients were summarized and discussed. RESULTS: Rupture of aortic aneurysm in both patients presented as a huge retroperitoneum haematoma by computed tomography scan. They were successfully saved by prompt body fluid compensation, emergency procedure, intraoperative resuscitation, and postoperative intensive care. CONCLUSIONS: Correct diagnosis, prompt surgical management, immediate intraoperative proximal aorta clamping during procedure, and effective management of postoperative complications were the key points to successful treatment of ruptured aortic aneurysm. PMID- 12903796 TI - Hyponatremia after transspheniodal surgery of pituitary adenoma. AB - OBJECTIVE: To clarify the frequency, presentation, associated factors, treatment and outcome of hyponatremia after transphenoidal surgery of pituitary adenomas. METHODS: Retrospectively reviewed the database of 183 patients who underwent transphenoidal surgery of pituitary adenoma between January 1999 and June 2000 in our department. RESULTS: 38.8% (71/183) had postoperative hyponatremia. Among them, 59.2% (42/71) appeared on the 4th to 7th day postoperatively. 59.2% (42/71) presented with nausea, vomiting, headache, dizziness, confusion and weakness. Hyponatremia was related to age, tumor size and adenoma type, but not related to sex and degree of resection. Treatment consisted of salt replacement and mild fluid restriction in 4 patients and salt and fluid replacement in 67 patients. Hyponatremia resolved within 16 days in all the patients. CONCLUSIONS: Hyponatremia often appeared about 7 days after transsphenoidal surgery of pituitary adenomas, especially in elderly and patients with macroadenomas and huge pituitary adenomas. The principle of treatment was salt and fluid replacement. PMID- 12903797 TI - Detection of telomerase activity in patients with mycosis fungoides. AB - OBJECTIVES: To detect telomerase activity in patients with mycosis fungoides (MF) and to study the role of telomerase in the tumorigenesis of MF. METHODS: The technique of PCR-ELISA was employed to detect telomerase activity in 35 patients with various stages of MF. RESULTS: 92.3% tumor stage of MF, 78.6% plaque stage of MF and 75.0% patch stage of MF had positive telomerase activity. The control samples had no telomerase activity. Telomerase activity in tumor stage of MF was significantly higher than that in plaque stage, while the latter was higher than that in patch stage. Telomerase activity was correlated with the stage of MF. CONCLUSION: High level of telomerase activity frequently occurred in patients with MF, suggesting that telomerase might play an important role in the tumorigenesis of MF and is a useful marker for the diagnosis of MF possibly. PMID- 12903798 TI - Regulating function of coenzymization and decoenzymization of the lactate dehydrogenase isozymes in the mouse tissues during hypoxia. AB - OBJECTIVE: To study the characteristics of changes of LDH enzyme patterns of mice under slight hypoxia. METHODS: Mice treated with artificial hypoxia, various tissues were made for the test of LDH enzymatic activity by the specific staining technique. LDH (1-5) relative percentage enzymatic activity (RPEA) were measured with CS-910 dual-wavelength thin layer chromatography scanner. RESULTS: The RPEA of LDH isozymes of various tissues after slight hypoxia shifted to the isozymes LDH1 and LDH2, whose principal subunits are H subunits, and the RPEA of LDH1 (H4), LDH2 (H3M) increased, while RPEA of LDH5 (M4) in various tissues decreased prominently except the cardiac muscle, and that of LDH4 (HM3) decreased as well. After polyacrylamide gel electrophoresis (PAGE) of the hypoxia treated cardiac muscle specimen was made, activity subbands originated regularly in the isoyme patterns of LDH, with the regularity of LDH1 (0 subband), LDH2 (0-1 subbands), LDH3 (0-2 subbands), LDH4 (1-3 subbands), LDH5 (2-4 subbands). After adding appropriate amount of NAD+ to the hypoxia treated cardiac muscle specimen, PAGE showed the subbands of four isoymes (LDH2-LDH5) reduced or even totally disappeared in the isozyme patterns. CONCLUSIONS: The negative feedback regulation of coenzymization and decoenzymization of LDH isozymes is one of the mouse stress responses to slight hypoxia. PMID- 12903799 TI - The value of telomerase assay in breast cancer. PMID- 12903800 TI - Vaccines as community-focused therapy. PMID- 12903801 TI - Melacine: an allogeneic melanoma tumor cell lysate vaccine. AB - Cancer vaccines have represented the holy grail of tumor immunology to many. In 2003, there are innumerable approaches to active specific immunotherapy for cancer. The identification of tumor antigens recognized by and able to activate human T-lymphocytes has heightened the enthusiasm for this approach. Melanoma is one of the diseases that has been primarily targeted by vaccine approaches. The identification of peptides and proteins associated with cancer has provided strategies to target specific components of the cancer cell. However, older vaccines utilizing products from whole cells may have a number of advantages. Melacine (Corixa Corp.) is an allogeneic melanoma tumor cell lysate combined with the adjuvant DETOX. Pioneered by Malcolm Mitchell, it has rare but confirmed antitumor activity in metastatic melanoma (5-10%). However, previous analysis of responses in advanced disease and a recent analysis in early-stage adjuvant trials suggest that Melacine may have its greatest benefit in a large subset of melanoma patients who express either the human leukocyte antigen (HLA) class I antigens A2 and/or HLA-C3. This finding must now be confirmed in a large perspective observation-controlled clinical trial to solidify the role of Melacine as an effective cancer vaccine in melanoma. PMID- 12903802 TI - Vaccination with heat shock protein-peptide complexes: from basic science to clinical applications. AB - There are currently over 150 medical centers worldwide enrolling patients in randomized, controlled Phase III clinical trials testing autologous cancer derived heat-shock protein (HSP)-peptide complexes for the treatment of renal cell carcinoma and melanoma. In addition, autologous HSP-peptide complexes have been or are being tested in Phase I and II trials of chronic myelogenous leukemia, lymphoma and pancreatic, gastric and colorectal cancers. The door has more recently opened to clinical testing of off-the-shelf HSP-based treatments for infectious diseases. This review recounts the long history of basic research on HSPs in immune response. A keen understanding of how these ancient molecules orchestrate the immune response to cancer and infections has been gained, providing a clear rationale for translating this knowledge into clinical medicine. PMID- 12903803 TI - Progress in prophylactic and therapeutic vaccines for human papillomavirus infection. AB - Virus-like particle (VLP) subunit vaccines composed of the major capsid protein L1 of the genital human papillomaviruses (HPVs) are now in Phase III clinical trials. The vaccines are immunogenic and safe and early results indicate efficacy. VLPs induce strong cell-mediated as well as humoral immune responses and chimeric VLPs including an HPV early protein may have therapeutic potential. Polynucleotide and recombinant viral vaccines encoding nonstructural viral proteins show therapeutic and prophylactic efficacy in animal models and are candidate immunotherapies for established low-grade benign genital infections. Vaccines designed to elicit cytotoxic T-lymphocytes specific for the HPV oncoproteins E6 and E7 show immunogenicity and efficacy in transplantable tumor models in rodents. In Phase I and II trials these vaccines are immunogenic and safe but show limited efficacy. PMID- 12903804 TI - Immunotherapy in multiple myeloma: current strategies and future prospects. AB - Multiple myeloma is still a fatal disease. Despite advances in high-dose chemotherapy supported by autologous transplantations, relapse of the underlying disease remains the primary cause of treatment failure. Strategies for post transplantation immunomodulation would be desirable for eradication of remaining tumor cells. Toward this end, immunotherapy aimed at inducing myeloma-specific immunity in patients has been exploited. Idiotype protein, secreted by myeloma cells, has been the main target for immunotherapy as it is the best-defined, tumor-specific antigen. The focus of this review article is the use of idiotype as a form of protein antigen to immunize patients, to load dendritic cells, or as part of DNA vaccines. Various strategies of immunotherapy and the outcome of clinical trials are also discussed. PMID- 12903805 TI - New approaches to the prevention and eradication of rabies. AB - Despite significant progress in improving the pre- and postexposure prophylaxis of human rabies, the development of better and more cost-effective vaccines and antiviral therapeutics remains a major goal for the treatment of human rabies, the control of animal rabies and particularly for the eradication of rabies virus reservoirs in terrestrial wildlife. In this review, we discuss the structural requirements for an effective rabies vaccine, as well as new strategies currently in use for the development of safer and more potent rabies vaccines for rabies prophylaxis and eradication. Finally, we discuss new immune therapeutics aimed at replacing the conventional administration of antirabies immunoglobulin used in rabies post-exposure prophylaxis in humans. PMID- 12903806 TI - Japanese encephalitis vaccines: moving away from the mouse brain. AB - Japanese encephalitis (JE) is a severe disease that is widespread throughout Asia and is spreading beyond its traditional boundaries. Three vaccines are currently in use against JE but only one is available internationally, a mouse-brain derived inactivated vaccine first used in the 1930s. Although this vaccine has been effective in reducing the incidence of JE, it is relatively expensive and has been linked to severe allergic and neurological reactions. Cell-culture derived inactivated and attenuated vaccines have been developed but are only used in the People's Republic of China. Other vaccines currently in various stages of development are DNA vaccines, a chimeric yellow fever-JE viral vaccine, virus like particle vaccines and poxvirus-based vaccines. Poxvirus-based vaccines and the chimeric yellow fever-JE vaccine have been tested in Phase I clinical trials. These new vaccines have the potential to significantly reduce the impact of JE in Asia, particularly if used in an oral vaccine delivery strategy. PMID- 12903807 TI - Bioinformatics: how it is being used to identify bacterial vaccine candidates. AB - Genomic sequencing has provided a tremendous amount of information that can be useful in vaccine target identification. The sheer volume of information available necessitates the use of new research disciplines and techniques. Using bioinformatics, researchers sift through available data to identify appropriate candidates for biological analysis. This review provides an overview of available bioinformatic techniques for vaccine candidate identification and a few examples of how these techniques are being applied to specific bacterial pathogens. PMID- 12903808 TI - Bacterial genomics and vaccine design. AB - Complete genome sequences are available for an increasing number of pathogenic bacteria. These new data are beginning to make an impact on the understanding of bacterial evolution and virulence. Thus far, however, vaccine development has had little benefit from genomics. Here we discuss how genomic sequence is being used in ways that could help identify useful bacterial antigens or create attenuated live vaccines. PMID- 12903809 TI - Alphavirus vectors for vaccine production and gene therapy. AB - Alphavirus vectors demonstrate high expression of heterologous proteins in a broad range of host cells. Replication-deficient as well as replication-competent variants exist. Systemic delivery of many viral antigens has elicited strong antibody responses in immunized mice and primates, and protection against challenges with lethal viruses was obtained. Similarly, prophylactic vaccination was established against tumor challenges. Attention has been paid to the engineering of improved targeting to immunologically active cells, such as dendritic cells. In the area of gene therapy, intratumoral injections of alphavirus vectors have resulted in potentially promising tumor rejection. Moreover, encapsulation of alphavirus particles into liposomes demonstrated efficient tumor targeting in mice with severe combined immunodeficiency, which permitted the initiation of clinical trials for patients with advanced kidney carcinoma and melanoma. PMID- 12903810 TI - Effects of genistein on invasion and matrix metalloproteinase activities of HT1080 human fibrosarcoma cells. AB - Effects of genistein on invasion and matrix metalloproteinase activities were investigated in HT1080 human sarcoma cells. Invasion of HT1080 cells through reconstituted basement membrane was inhibited when the cells were treated with 100 micromol/L and 200 micromol/L genistein. At the same concentrations, genistein not only suppressed latent forms of matrix metalloproteinase-2 and -9 (MMP-2 and MMP-9) to convert into active forms, but also increase dramatically the tissue inhibitor of metalloproteinase (TIMP-1) mRNA contents and reverse the imbalance of MMPs and TIMPs. However, expressions of MMP-2 and MMP-9 were not significantly affected. Suppression of MMP activation and increase of TIMP-1 expression will decrease matrix degradation by MMPs, and consequently inhibit invasions of the cells. These results emphasized the existence of the imbalance between MMPs and TIMPs in tumor invasion and metastasis formation. The value of genistein as a drug for antiinvasion and anti-metastasis chemotherapy was suggested. PMID- 12903811 TI - p53 gene mutations in non-small cell lung cancer detected by polymerase chain reaction single-strand conformation polymorphism analysis. AB - Mutations of the p53 tumor suppressor gene are the most frequent genetic alterations detected in human lung cancer. To assess the pathogenic significance of p53 gene alterations in Chinese non-small cell lung cancer (NSCLC), 74 paired samples of primary lung cancer and normal lung tissue far away from the cancer were analyzed for mutations of the p53 gene (exons 5-8) using exon-specific PCR, single-strand conformation polymorphism (PCR-SSCP). p53 mutations were observed in 55.4% (41/74) of the samples. No linkages were detected between the incidence of p53 mutations and histological type, lymph node metastasis, age or sex. Significant association between p53 mutations and degree of differentiation in adenocarcinomas, not in squamous cell carcinomas, was observed. The frequency of p53 mutations in smokers (65.3%) was higher than in nonsmokers (33.3%) and reached statistical significance. We also found p53 mutations in 6/7 samples which had tissue invasion and distant metastasis. These results suggest that smoking could be an important factor in lung carcinogenesis, p53 mutation is a worse prognosis indicator in adenocarcinomas and related to high aggressive behavior of human lung cancer. PMID- 12903812 TI - Inhibition of farnesyl protein transferase and P21ras memebrane association by d limonene in human pancreas tumor cells in vitro. AB - The monoterpene d-limonene inhibit the plasma-membrane associated P21ras expression and the post-translational isoprenylation of P21ras, a mechanism that may contribute to its efficacy in the chemoprevention and therapy of chemically induced rodent cancers and some human solid tumor cells. In the present study, the relative abilities of d-limonene to inhibit membrane associated P21ras expression in pancreas tumor cell (PaCa) was carried out with Western blotting, and the inhibition of farnesyl protein transferase (FTPase) activity during the Ras protein isoprenylation and cell proliferation were determined. Concomitantly, the effects of d-limonene on P21ras localization by immunohistochemistry and H ras oncogene expression in PaCa tumor cell line by Northern blotting were observed. The results showed that d-limonene inhibited FPTase activity, thus to reduce P21H-ras isoprenylation. d-limonene could decrease P21ras membrane association and increase cytosolic accumulation of P21ras. This phenomenon was also noted when d-limonene-treated PaCa cells were stained immunohistochemically with anti-P21ras antibody. It is suggested that the inhibition of FPTase activity was closely related with the inhibiton of P21ras membrane association and the alteration of P21ras localization. Inhibition of farnesylation of P21ras altered their intracellular localization and, hence, disrupted their biological activity, but no relationship with H-ras oncogene expression was found. PMID- 12903813 TI - The correlativity analysis of six methods of detecting apoptosis. AB - The aim of this study was to compare six methods of detecting apoptosis induced by extracellular adenosine triphosphate (ATP) in human leukemic lymphocytes with purinergic P2Z receptors. These methods used were electron microscopy (EM), detection of internucleosomal DNA fragmentation by agarose gel electrophoresis, autoradiographic analysis of DNA fragmentation, in situ labeling of DNA strand breaks with fluorescein dUTP and exogenous terminal deoxynucleotidyl transferase (TUNEL), quantitation of 3'-ends of DNA breaks by labeling with alpha32PdCTP(TdT assay), and quantitation of apoptotic cells with fluorescein-annexin V using flow cytometry (FCA). We found EM and detection of DNA ladder pattern by agarose gel electrophoresis to b specific, but lacking in sensitivity. The combination of autoradiography and gel electrophoresis gave an increase in sensitivity of at least 50 fold although, of all the methods, the TdT assay was shown to be most sensitive. The four methods for quantifying apoptosis-EM, FCA, TUNEL and TdT assay proved to be reliable and gave statistically similar results on apoptotic lymphocytes. These observations indicate it is essential to combine specific, sensitive and quantitative techniques in detecting apoptosis. PMID- 12903814 TI - HBV X protein (HBX) interacts with general transcription factor TFIIB both in vitro and in vivo. AB - OBJECTIVE: In order to demonstrate the binding of HBV X protein (HBX) with the general transcription factor TFIIB. METHODS: In vitro glutathion S-transferase (GST) resin Pull-Down assay and Far-Western Blotting assay, in vivo Co immunoprecipition assay were used. RESULTS: The X199 (51-99) domain of HBX is reponsible for HBX binding to TFIIB. While the d10 domain (125-295) of TFIIB is required for TFIIB binding to HBX. When the two basic amino acids (K) at position 178 and 189 of TFIIB were substituted by neutral amino acids (L), the binding of TFIIB K178L and K189L to HBX was siginificantly reduced. When the the basic amino acids were substituted by the acidic amino acids (E), the binding of TFIIB K178E and K189E to HBX were almost lost. In vitro results of HBX binding to TFIIB were further confirmed by in vivo co-immunoprecipitation assay. Our results also indicated that the Woodchuck hepatitis virus X protein (WHX) interacts with TFIIB. CONCLUSION: These results suggested that the communication between HBX and general transcription factor TFIIB is one of the mechanisms which account for its transcriptional transactivation. PMID- 12903815 TI - CD28/B7-mediated costimulation is required for parathyroid gland allograft rejection in rats. AB - The activation of T cells to differentiate and to proliferate is an essential step in the immune response to antigen, especially in cell mediated acute allograft rejection. Besides the interaction of CD3/TCR complex with Ag/MHC complex presented on antigen-presenting cells, a complete T-cell activation and proliferation requires a second costimulatory signal. The interaction of CD28/CTLA-4 and B7 is a major costimulatory pathway for T Cell activation. Inhibition of this pathway results in development of antigen-specific unresponsiveness and clonal anergy. In present study, the biologic function of anti-CD28 monoclonal antibody and its Fab fragment were investigated in vitro and in vivo. The results indicate that mAbCD28 and its Fab fragments could promote the functional recovery of allografts and prolong the graft survival, but could not reverse the acute rejection or induce transplantation tolerance in the rat PTG allograft model. We also found that peripheral TNF-alpha level and NK-cell activity were suppressed in the presence of mAbCD28 and its Fab fragments for a relatively long time after PTG transplantation. PMID- 12903816 TI - A diagnostic significance of early renal impairment with liver cirrhosis through the determination of urinary enzymes. PMID- 12903817 TI - Expression of ICAM-1 and LFA-1 molecules in relation to renal allograft rejection in rats. AB - OBJECTIVE: The purpose of this study was to assess the renal graft expression of ICAM-1 (intercellular adhesion molecule-1) and LFA-1 (lymphocyte function associated antigen-1) molecule with relation to graft rejection. METHODS: Rat kidney transplantation was performed according to the procedure of Kamada with some modification. Experimental rats were divided into 5 groups. The survival time of recipient rats and function of grafts after renal transplantation were observed. The sections of renal graft were stained for monoclonal antibody ICAM-1 and LFA-1, and then quantification of ICAM-1 and LFA-1 expression was accomplished by computer image analysis. RESULTS: ICAM-1 and LFA-1 increased significantly in the renal allograft rejection group as compared with the non rejection groups (P<0.05). CONCLUSION: Both biopsy of renal graft and monitoring of ICAM-1 and LFA-1 are useful tools in diagnosing and treating acute rejection. PMID- 12903818 TI - Feasibility study of an ultrasound contrast agent (levovist) in color Doppler imaging of liver neoplasms. AB - The purpose of this study was to determine the efficacy of using an ultrasound contrast agent (levovist) to enhance the color Doppler imaging of liver neoplasms. Thirty patients with hepatic tumors were enrolled in this study. After intravenous administration of levovist, the color Doppler signals of normal hepatic vessels were enhanced. In various hepatic tumors, the different patterns of tumor vascularity were observed, which had not been demonstrated in conventional non-contrast color Doppler imaging. In 11 of 16 patients with hepatocarcinoma, additional color Doppler signals were observed in the central part of the tumors. On the contrary, 3 patients with metastatic liver lesions the enhanced color Doppler signals appear only at the peripheral of tumors. A typical rim-like color enhancement was seen in 2 of the 3 cases. In six patients with hepatic hemangiomas contrast-enhanced color Doppler imaging demonstrated the blood vessels at the margin of the neoplasms. Contrast-enhanced color Doppler imaging improves the visualization of the hepatic neoplasm vascularity. This technique holds great promise for detecting small liver tumors and differentiating hepatic neoplasms. PMID- 12903819 TI - Intraoperative ultrasound for hepatic neoplasm during surgery. AB - OBJECTIVE: The purpose of this study was to determine the impact of intraoperative ultrasound (IOUS) on the management of patients with neoplasms of the liver. METHODS: Forty-nine patients operated on for liver or other pathologic processes were examined intraoperatively with 5.0 MHz special ultrasound transducers during surgical exploration of the abdomen. Subjects were evaluated because of known or suspected disease of the liver. Preoperative imaging studies included percutaneous ultrasound (n = 49), magnetic resonance imaging (n = 11), and computed tomography (n = 34). Intraoperative evaluation on all patients included inspection, bimanual palpation, and ultrasonography. Comparison between preoperative imagings and IOUS were analysed. RESULTS: Sensitivity for detection of hepatic neoplasms showed in intraoperative ultrasound, percutaneous ultrasound, magnetic resonance imaging and computed tomography as 100% (23/23), 74% (17/23), 74% (14/19) and 75% (6/8). Specificity showed 100% (26/26), 100% (26/26), 93% (14/15) and 67 (2/3). In seven patients (14%), the neoplasms were not found by inspection, bimanual palpation, and identified only by IOUS. CONCLUSIONS: Intraoperative ultrasound is the most sensitive and specific method for detection and surgery of liver neoplasms, especially the occult neoplasms and small size lesion (< 2 cm). PMID- 12903820 TI - Failure and complication following surgical treatment of scoliosis--an analysis of 101 cases. AB - Complications occur frequently after surgical treatment of scoliosis. In order to prevent from them effectively, 101 cases with failure and postoperative complications were analysed. They included rod fracture in 22 cases (15 Harrington rods, 4 Zielke rods, and 3 Luque rods); recurrence of curve severity in 12 cases; broken or loossened luque wires in 15 cases; loss of thoracic kyphosis (flat back) in 6 cases; progressive kyphosis with or without paraplegia following incorrect posterior decompression in 5 cases; and increased unbalance of shoulders after instrumentation in 2 cases due to neglect of the tilting of the first thoracic vertebra. Infection occurred in 8 cases (incision infection 7 cases, deep wound infection in 1 case); and pneumothorax in 1 case. They were induced by biomechanical factors in 23 cases (22.8%), incorrect selections of indications in 29 cases (28.7%), operational mistakes in 37 cases (36.6%), internal fixation factors in 15 cases (14.9%). The authors hold that there are quite a lot of factors leading to occurrence of complications and the effective way for prevention from them is to understand the factors and main technical points related to internal fixation. PMID- 12903821 TI - Effects of exogenous nitric oxide on the function of isolated working hearts. PMID- 12903822 TI - Clinical and experimental studies of large amplitude action potential of the suffered facial muscles in intratemporal facial nerve paralysis. AB - OBJECTIVE: To testify the phenomenon that large amplitude action potential appears at the early stage of facial paralysis, and to search for the mechanism through clinical and experimental studies. PATIENTS (ANIMALS) AND METHODS: The action potentials of the orbicular ocular and oral muscles were recorded in 34 normal persons by electromyogram instruments. The normal range of amplitude percentage was found out according to the normal distribution. One hundred patients with facial paralysis were also studied. The action potentials of facial muscles were recorded in 17 guinea pigs before and after the facial nerve was compressed and the facial nerve was examined under electromicroscope before and after the compression. RESULTS: The amplitude percentage of the suffered side to the healthy side was more than 153 percent in 6 of the 100 patients. Large amplitude action potential occured in 35 per cent guinea pigs which were performed the experiment of facial nerve compression. Electromicroscopic examination revealed separation of the lammae of the facial nerve's myelin sheath in the guinea pigs which exhibited large amplitude action potential. CONCLUSION: The facial nerve exhibited a temporary over-excitability at the early stage of facial nerve injury in some patients and guinea pigs. If the injury was limited in the myelin sheath, the prognosis was relatively good. PMID- 12903823 TI - Nitric oxide synthase mRNA expression and localization in pulmonary artery of hypoxic pulmonary hypertensive rats. PMID- 12903824 TI - The diagnosis and treatment of gastrinoma: report of 17 cases. AB - OBJECTIVE: To study the experience of diagnosis and treatment of gastrinoma. METHODS: A retrospective study of 17 patients with gastrinoma seen from 1978 through 1998 in PUMC hospital. RESULTS: Three of the 17 cases were associated with multiple endocrine neoplasia type I (MEN-I) syndrome. Tumors dispersed extensively in pancreas (31%), duodenum (25%), lymphnode (19%) and other place (25%). Ultrasonography and computed tomography (CT) were sensitive in localizing tumor in pancreas and liver, but were not so well for small tumor in duodenum and lymph node. The angiography, percutaneous transhepatic portal sampling (PTPS), scintigraphy and endoscopic ultrasonography (EUS) also can be used. Sixteen of 17 patients underwent 38 operations and 56 percent of the 16 cases underwent several operations. Seven patients at last performed total gastrectomy that was still considered as a choice of treatment. Tumor resection was rare because the advancing of tumor in most cases. CONCLUSIONS: Diagnosing gastrinoma in its early stage and striving for the tumor resection and reducing the metastasis of liver were the cruxes for good prognosis. PMID- 12903825 TI - A new Pvu II restriction site of mitochondrial DNA in a patient with mitochondrial encephalomyopathy. PMID- 12903826 TI - Effects of alfentanil and esmolol on hemodynamic and catecholamine response to tracheal intubation. AB - OBJECTIVE: To compare the effects of alfentanil and esmolol on hemodynamic and catecholamine response to tracheal intubation. METHODS: Thirty-five adult patients were randomly allocated to one of three groups, Group A (control group). Group B (esmolol group) and Group C (alfentanil group). The patients received either 2 mg/kg esmolol (in Group B) or 30 microg/kg alfentanil (in Group C) before intubation. Tracheal intubation was performed with 4 mg/kg thiopental and 0.1 mg/kg vecuronium and 3% isoflurane. Systolic blood pressure (SBP), diastolic blood pressure (DBP), mean blood pressure (MBP), heart rate (HR), norepinephrine (NE), epinephrine (E) and dopamine (DA) were measured before and after intubation. RESULTS: The control group had a baseline SBP of 149 +/- 23 mmHg while Groups B, C had a baseline SBP of 148 +/- 23, and 150 +/- 21 mmHg, respectively (P>0.05). Three min after tracheal intubation, the control group SBP increased to 160 +/- 30 mmHg and Group B remained at the baseline level, 147 +/- 5 mmHg, and Group C significantly decreased to 91 +/- 22 mmHg (P<0.01). Two min after intubation HR in Group B increased significantly but 3 min after intubation HR in Groups B and C were significantly lower than that of control group (P<0.05). NE in Groups A and B increased significantly to 5.75 +/- 3.51 and 6.75 +/- 3.30 nmol/L 3 min after intubation (P<0.01). In Group C, 3 min after intubation NE was not significantly different from the baseline but E decreased significantly (P<0.01). CONCLUSION: 2 mg/kg esmolol can moderate the hemodynamic response to tracheal intubation to a certain extent and 30 microg/kg alfentanil can completely attenuate the hemodynamic and catecholamine responses. PMID- 12903827 TI - Cloning of encoding sequences for chemokine receptors CXCR4 and CCR5 from a Chinese lymphocyte cDNAs. PMID- 12903828 TI - The ionic basis of the abnormal electrophysiological features of the myocytes isolated from the right ventricle of chronic hypoxia guinea pigs. PMID- 12903829 TI - Role of isoflavones in the hypocholesterolemic effect of soy. AB - Epidemiologic data suggest an inverse relationship between the consumption of soy isoflavones and cardiovascular disease risk. The aims of this review are to determine if isoflavones play a role in the hypocholesterolemic effect of soy and whether the studies realized with that scope were adequately designed. In humans, most studies have been performed in postmenopausal women. The results are inconsistent, however; some studies show a decrease in total cholesterol and low density lipoprotein concentrations, and an increase in high-density lipoprotein levels, and other investigations fail to show any beneficial effect of soy isoflavones on lipid profiles. In most studies, beneficial effects could not be attributed with certainty to soy isoflavones. If these components have any health protecting effect in humans, it is small in comparison with the effect of soy protein itself. There are currently not enough data to recommend the consumption of isoflavone supplements to lower plasma cholesterol levels. PMID- 12903830 TI - Bioactive isoflavones in functional foods: the importance of gut microflora on bioavailability. AB - Isoflavones present in soy may have risk and benefits to human health. Human gut microflora have been shown to exert metabolic activities on isoflavones, which influences bioavailability and bioactivity. Absorption of isoflavones is likely to occur in the small intestine where there is a diverse range of microfloral species able to hydrolyze conjugated isoflavones, releasing the bioactive aglycone for absorption or further metabolism and reconjugation. The identification of gut microbes that metabolize isoflavone aglycones is not well established. Such information may lead to a better understanding of the bioavailability of isoflavones in functional foods. PMID- 12903831 TI - Finding a place for immunonutrition. AB - The role of certain nutrients that seem to have pharmacologic effects on immune and inflammatory parameters has been studied over the last two decades. This area of research is called immunonutrition. A recent study administered a combination of immunonutrients perioperatively in well nourished gastrointestinal cancer patients undergoing elective surgery. The rate of infectious complications and length of hospital stay was decreased in those receiving immunonutrients compared with patients receiving no nutritional support. The role of the individual components of this immunonutrition and the clinical trials of similar combinations in surgical gastrointestinal cancer patients are reviewed. PMID- 12903832 TI - Usefulness of artificial sweeteners for body weight control. AB - Artificial sweeteners provide lower calorie food products but have been thought to increase appetite and perhaps lead to increased body weight compared with high sucrose foods. A recent trial demonstrated that subjects supplementing their diets with artificially sweetened foods lost weight, whereas those consuming high sucrose foods gained weight. This review sheds some light on the controversy regarding the effects of artificial sweeteners in body weight control. PMID- 12903833 TI - Lactose intolerance: a self-fulfilling prophecy leading to osteoporosis? AB - Symptoms of lactose intolerance are unlikely to occur under usual dietary conditions. Yet, self-described "lactose-intolerant" individuals often restrict dairy and calcium intake. A new study suggests that such individuals have reduced peak bone mass and increased incidence of osteopenia, and are at greater risk of osteoporosis and bone fractures. PMID- 12903834 TI - Conditional gene targeting. AB - Precise chromosomal alterations can be made in restricted tissues at chosen times in the living experimental animal. The engineering of such conditional systems has been successfully tackled in transgenic mice during the last decade. This review focuses on how several different genetic regulatory elements from lower organisms can be applied in combination to achieve this goal. PMID- 12903835 TI - Molecular biologic aspects of cartilage and bone: potential clinical applications. AB - The formation of cartilage and bone tissue from condensing mesenchymal stem cells is taking place in the early stage embryo, but also in the growth plate and during fracture repair in adults. Resident mesenchymal stem cells have been identified in bone marrow, periosteum, and in muscles. These pluripotent cells are attractive as therapeutic cells in cartilage and bone reconstructive procedures, since they can differentiate into chondrocytes and osteoblasts upon appropriate stimuli, such as certain growth factors. Members of the transforming growth factor-beta (TGF-beta) superfamily, including TGF-beta1-3, bone morphogenetic proteins (BMPs), and activins, are essential mediators in cell proliferation and differentiation and play regulatory roles in cartilage and bone formation. This review presents functional data on TGF-beta1 in cartilage formation, BMP2 and BMP3 in bone formation, and a role of the BMP-regulatory protein noggin in BMP2 processing. Potential clinical applications of these proteins are further demonstrated and discussed. PMID- 12903836 TI - Involvement of the proteasome in IL-1beta induced suppression of islets of Langerhans in the rat. AB - The cytokine IL-1beta suppresses rodent islets of Langerhans in vitro. Presently we used inhibitors of the proteasome to investigate if these compounds could counteract the suppressive effects of the cytokine. Thus, isolated rat islets were cultured and pre-treated with proteasome inhibitors and subsequently exposed for 48 h to 25 U/ml human IL-1beta. After this period functional tests were carried out. The rate of glucose oxidation (pmol/10 islets x 90 min) was suppressed by IL-1beta (115 +/- 17 vs. control 380 +/- 57). Pre-treatment with 10 microM of the proteasome inhibitor MG115 (N-carbobenzoxyl-leu-leu-norvalinal) and 100 microM of the calpain inhibitor norLEU (N-acetyl-leu-leu-norleucinal; known to affect proteasome activity) counteracted the suppressive effects (253 +/- 17 and 262 +/- 10 respectively). The calpain inhibitor alIMET (N-acetyl-leu-leu methional) had no effect. MG115 (10 microM) and norLEU (100 microM) blocked nitric oxide formation induced by IL-1beta, while alIMET was without effect. We also investigated if IL-1beta could influence the expression of two inducible proteasome subunits, namely LMP2 and LMP7, and found that the cytokine increased the mRNA expression of the proteasome subunit LMP2 in islets, and that the proteasome inhibitor MG115 prevented this increase. In conclusion our study shows that IL-1beta increases the transcription of the proteasome subunit LMP2, and that the proteasome is involved in IL-1beta induced suppression of islet function. Moreover, the observation that inhibitors of the proteasome protect islets against IL-1beta induced inhibition of glucose metabolism, suggests that these compounds might be worthwile to explore in future therapies against the development of type 1 diabetes. PMID- 12903837 TI - High sensitivity CRP and serum amyloid A as expressions of low grade inflammation do not correlate with bFGF or VEGF. AB - High sensitivity CRP (hsCRP) and serum amyloid component A (SAA) are two acute phase proteins that have been shown to be prognostic markers in cardiovascular diseases. Vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) are two angiogenic growth factors involved in the growth and invasion of solid tumors and hematological malignancies. They also play an important role in cardiovascular and inflammatory diseases. The acute phase response induces the production of angiogenic peptides and angiogenic peptides have also been shown to induce the synthesis of inflammatory cytokines. HsCRP and SAA, bFGF and VEGF in serum and EDTA plasma were measured in samples from 80 human blood donors. There were strong correlations between hsCRP and SAA but there were no significant correlation between hsCRP or SAA and any of the angiogenic peptides. Low-grade inflammation did not induce bFGF or VEGF production. PMID- 12903838 TI - Perspectives on hepatitis B infections and the efficacy of vaccination (hepatitis B and pneumococci) in dialysis patients. AB - Hepatitis B is a well known problem in dialysis units. We therefore examined the historical frequency of hepatitis B carriers in our unit, our vaccination program to hepatitis B virus (HBV), the response to hepatitis B vaccine, the IgG subclass response of anti-HBs and the response and IgG subclass response to pneumococcal vaccination (another vaccine) in dialysis patients. From 1970 and onwards 23 HBV carriers were found, but no new cases of hepatitis B occurred during the study period, i.e. from 1980 and onwards. Only one of the carriers was alive by the end of 2001. In four patients liver disease (in one of them liver cirrhosis) may have been a concomitant cause of death. The antibody response to hepatitis B vaccine was significantly lower in patients than in staff. In four patients a fourth injection was cancelled due to transplantation and bad health, while such data were lacking in 8 cases. In anti-HBs positive patients and controls a significant difference in the response of healthy adults was observed in anti-HBs IgG1 (p < 0.001) vs all other IgG subclasses. Dialysis patients had low levels, or negative findings, in all cases, with IgG1 as the highest proportion found (3/11 patients). An antibody response to pneumococcal vaccination was registered in 25 out of 29 dialysis patients (in all 86%). The IgG-subclass vaccination response to pneumococci in 28 dialysis patients was mainly IgG2 and IgG1 but also occurred in IgG3 and IgG4. Prevaccination antibody levels of the controls were higher in IgG1 and IgG2 (p < 0.01) (n = 21) than in dialysis patients (n = 28). Hepatitis B is nowadays a rare, but still dangerous disease in nephrology units. Dialysis patients have a reduced response to hepatitis B vaccine and vaccination schedules should be started early as some patients otherwise may not receive a fourth injection. The adequate antibody response to pneumococcal vaccination mainly due to IgG2 and IgG1 antibodies indicates that the antigen involved is important in vaccination responses in dialysis patients. PMID- 12903839 TI - Genetic susceptibility, predicting risk and preventing cancer. AB - A polygenic approach to disease prevention has become a realistic goal that has arisen from the sequencing of the human genome. Some believe it will be possible to identify individuals as susceptible by their genotype and to prevent disease by targeting interventions to those at risk. However, doubts have been expressed about the magnitude of these genetic effects, and hence the potential to apply them either to individuals or to populations. Published data suggest that the familial aggregation of breast cancer not due to the known high penetrance genes is polygenic, which implies that the distribution of risk in the population is continuous. This model is likely to apply to other common cancers. The utility of a continuous distribution for identifying a high-risk group of the population for targeted preventive intervention depends on the spread of that risk distribution. For breast cancer, the data are compatible with a log-normal distribution of genetic risk in the population which is sufficiently wide to provide useful discrimination of high- and low-risk groups. Assuming all the susceptibility genes could be identified, the half of the population at highest risk would account for 88% of all cases. In contrast, if currently identified risk factors for breast cancer were used to stratify the population, the half of the population at highest risk would account for only 62% of all cases. These results suggest that in the future the construction and use of genetic risk profiles may provide significant improvements in the efficacy of population-based programmes of intervention for cancers and other diseases. PMID- 12903840 TI - Novel approaches to identify low-penetrance cancer susceptibility genes using mouse models. AB - Studies of cancer predisposition have largely concentrated on the role of high penetrance susceptibility genes. Less than 10% of the total human tumor burden, however, is accounted for by mutations in these genes. More genetic variation in cancer risk is likely to be due to commoner but lower penetrance alleles. In man, such modifier genes will be difficult to find since they do not segregate as single Mendelian traits. The mouse offers a powerful system for studying polygenic traits such as cancer and has been widely used for this purpose. Novel approaches that might accelerate the identification of these low-penetrance cancer susceptibility genes by using mouse models will be discussed. PMID- 12903841 TI - Development of novel selective cell ablation in the mammary gland and brain to study cell-cell interactions and chemoprevention. AB - We have generated transgenic mice which express the gene encoding Escherichia coli nitroreductase (NTR) specifically in the luminal epithelial cells of the mammary gland and the glial cells of the brain. The enzyme activates an antitumour drug CB 1954, to produce a cross-linking agent that kills all cells expressing the enzyme. We have shown that administration of the antitumour drug CB 1954 rapidly and selectively kills these cells. Original experiments demonstrated the ability to ablate the luminal cells in the mammary gland with no apparent bystander effect. Subsequently, astrocytes expressing nitroreductase under the targeting of the GFAP promoter were selectively ablated following administration of the prodrug CB 1954 produces a degeneration of granular neurones due to changes in glutamate levels. Recent experiments demonstrated inhibition of myc-dependent mammary tumours using the same enzyme (nitroreductase)-prodrug (CB 1954), combination. Owing to the ease of control of NTR-mediated cell ablation, we anticipate that this system will supersede herpes simplex virus type 1 thymidine kinase. There are widespread potential applications for this approach in the dissection of complex cellular interactions during development and in the adult organism. The present transgenic models also have important applications for the study in vivo of novel prodrugs that can be selected for variable degrees of bystander effects. Such studies will have particular significance for those groups advocating the use of NTR as an appropriate enzyme for gene-directed enzyme prodrug therapy by providing models of a wide range of human disease for mechanistic and therapeutic experimentation. The results clearly demonstrate that the model has potential to study chemoprevention and fundamental questions on cell-cell interactions in cell biology. PMID- 12903842 TI - Mouse skin as a model for cancer chemoprevention by nonsteroidal anti inflammatory drugs. AB - The mouse skin model of multistage carcinogenesis has demonstrated that cancer results from a synergism between genotoxic and nongenotoxic factors. The former induce irreversible genetic alterations, whereas the latter promote tumor development by favoring the clonal outgrowth of the genetically altered cells. While therapeutic gene repair is a still unrealized dream, tumor promotion provides an attractive target for cancer prevention. A key event in epithelial tumor development is an aberrant constitutive overexpression of cyclooxygenase-2 (COX-2), being detectable already in premalignant lesions and leading to an overproduction of prostaglandins. In the mouse skin model, prostaglandin F2alpha has been identified as an endogenous tumor promoter. The well-established chemopreventive effect of nonsteroidal anti-inflammatory drugs seems to be mainly due to COX-2 inhibition. Targeted transgenic overexpression of COX-2 in mouse epidermis induces a preneoplastic phenotype and renders the tissue extremely sensitive to genotoxic carcinogens; i.e., for the induction of skin tumor development, tumor promoter treatment can be omitted in those animals. It is concluded that COX-2 acts as an endogenous tumor promoter and that its overexpression represents a first order risk factor for cancer development. Conversely, specific COX-2 inhibitors rank among the most promising agents for cancer chemoprevention. PMID- 12903843 TI - Preclinical models for chemoprevention of colon cancer. AB - Colon cancer is the second leading cause of cancer incidence and death in the USA in 2002. Specific genetic defects have been identified which cause hereditary colon cancers in humans. In addition, a number of intestinal luminal risk factors for colon cancer have been described. This information has been exploited to develop experimental cell and rodent models which recapitulate features of human colon cancer. In this chapter, we will discuss the strengths and limitations of these models to further our understanding of basic mechanisms of colon carcinogenesis and to develop strategies for colon cancer chemoprevention. PMID- 12903844 TI - New cancer biomarkers deriving from NCI early detection research. AB - Cancer is not a single disease but an accumulation of several events, genetic and epigenetic, arising in a single cell over a long time interval. A high priority in the cancer field is to identify these events. This can be achieved by characterizing cancer-associated genes and their protein products. Identifying the molecular alterations that distinguish any particular cancer cell from a normal cell will ultimately help to define the nature and predict the pathologic behavior of that cancer cell. It will also indicate the responsiveness to treatment of that particular tumor. Understanding the profile of molecular changes in any particular cancer will be extremely useful as it will become possible to correlate the resulting phenotype of that cancer with molecular events. Achieving these goals and knowledge will provide an opportunity for discovering new biomarkers for early cancer detection and developing prevention approaches. This will also help us identify new targets for therapeutic development. Advancement in technology includes methods and tools that enable research including, but not limited to, instrumentation, techniques, devices, and analysis tools (e.g., computer software). Resources such as databases, reagents, and tissue repositories are different than technologies. The identification and definition of the molecular profiles of cancer will require the development and dissemination of high-throughput molecular analysis technologies, as well as elucidation of all of the molecular species embedded in the genome of cancer and normal cells. The main challenge in cancer control and prevention is to detect the cancer early. This could then enable effective interventions and therapies contributing to reduction in mortality and morbidity. At a specific time, biomarkers serve as molecular signposts of the physiologic state of a cell. These signposts are the result of genes, their products (proteins) and other organic chemicals made by the cell. Biomarkers could prove to be vital for the identification of early cancer and subjects at risk of developing cancer as a normal cell progresses through the complex process of transformation to a cancerous state. This chapter discusses ongoing research in genetic and proteomic approaches to identify molecular signatures such as protein profiles, microsatellite instability, hypermethylation, and single nucleotide polymorphisms. Other topics covered here include the use of genomics and proteomics as high-throughput technology platforms to facilitate biomarker-aided detection of early cancer. Other areas covered include issues surrounding the analysis, validation, and predictive value of biomarkers using such technologies. Recent advances in noninvasive techniques, such as buccal cell isolates serving as viable sources of biomarkers, complementary to traditional sources such as serum or plasma, are also presented. The review also brings attention to the efforts of the Early Detection Research Network (EDRN) at the National Cancer Institute (NCI), in bringing together scientific expertise from leading national and international institutions, to identify and validate biomarkers for the detection of precancerous and cancerous cells in determining risk for developing cancer. The network's serious determined efforts in linking discovery to process development, resulting in early detection tests and clinical assessment, are also discussed. PMID- 12903845 TI - Tamoxifen's impact as a preventive agent in clinical practice and an update on the STAR trial. AB - Tamoxifen has long been an established adjuvant treatment in the management of advanced breast cancer. Studies carried out by the National Surgical Adjuvant Breast and Bowel Project (NSABP) and other groups also established its use in the treatment of early breast cancer and subsequently as a risk-reducing agent for the development of contralateral breast cancer in breast cancer patients. The NSABP began its investigation of tamoxifen as a preventive agent in the early 1990s in women who had never had the disease with its Breast Cancer Prevention Trial (P-1). Its second prevention study, the STAR trial, is currently under way. PMID- 12903846 TI - Aromatase inhibitors in prevention--data from the ATAC (arimidex, tamoxifen alone or in combination) trial and the design of IBIS-II (the second International Breast Cancer Intervention Study). AB - Current prevention trials have shown that tamoxifen can reduce the incidence of breast cancer by about 30%-40% in high-risk women, but that the risk of thromboembolic disease and endometrial cancer are increased about twofold. An alternative approach for postmenopausal women is to use an aromatase inhibitor to reduce oestrogen to very low levels. Data from the ATAC adjuvant trial indicate that the aromatase inhibitor anastrozole is more effective than tamoxifen in reducing recurrence and preventing new contralateral tumours, and also has a more favourable side-effect profile. This has led us to launch a new prevention trial- IBIS-II--in 6,000 high-risk postmenopausal women comparing anastrozole against placebo. A parallel trial will compare anastrozole against tamoxifen in 4,000 women with locally excised DCIS. PMID- 12903847 TI - HRT opposed to low-dose tamoxifen (HOT study): rationale and design. AB - The rationale for the HOT study is mainly based on the findings of the Italian Tamoxifen Prevention Study, where 5,408 healthy hysterectomized women aged 35-70 years were randomized to 20 mg/day of tamoxifen or placebo for 5 years. After 81.2 months median follow-up, 79 breast cancers occurred (34 on tamoxifen versus 45 on placebo, p=0.215). In the subgroup of 1,580 women who used estrogen replacement therapy (ERT) at some point during the study, 23 breast cancers were observed: 17 on placebo and 6 on tamoxifen (hazard ratio=0.35, 95% CI, 0.14 0.89). Pharmacokinetic and pharmacodynamic (surrogate endpoint biomarkers) studies showed that a lower dose of tamoxifen (such as 5 mg/day) does not affect the drug's activity on several biomarkers of both cardiovascular and breast cancer risk. We therefore propose a multicenter placebo-controlled phase III trial in postmenopausal healthy women on hormone replacement therapy (HRT) to assess whether the combination of HRT and low-dose tamoxifen retains the benefits while reducing the risks of either. PMID- 12903848 TI - Is mammography screening for breast cancer really not justifiable? AB - The consensus that breast screening is effective for women aged 50-59 years was shattered when Gotzsche and Olsen suggested that there are no reliable data to support screening by mammography. In practice, their concerns have been difficult to address because for many studies purporting to show effectiveness, adequate data have not been published to confirm that they were valid. Further, the trials in Canada for which such data are available did not show effectiveness of screening mammography. Since the 1990s, there has been an unprecedented reduction in breast cancer mortality in many countries. However, the reductions have no clear link to screening, but are probably due to the implementation of adjuvant treatment with chemotherapy and tamoxifen. Whether screening will have an additional impact in the future is unclear. After reviewing the published evidence, I conclude that the additional contribution of mammography over screening by good breast physical examinations and breast self examination is to detect good prognosis breast cancers, as the benefit of screening derives from the earlier detection of relatively advanced breast cancers, providing good therapy is given. If women choose mammography screening, they should understand that their risk of dying in the next few years may not be reduced. PMID- 12903850 TI - Political interpretation of scientific evidence--case study of breast cancer screening policies around the world. AB - Early detection of breast cancer has been studied for effectiveness in reducing mortality for more than four decades. During this period numerous studies took place, with more than half a million participating women. In spite of large amounts of nonconflicting data, different countries took varying lengths of time to establish a policy, and came up with a variety of policies, quite different from one another. Inherent differences between the countries in structure of the health system, in the commitment to public health activities, and in opinions and health habits of the relevant populations may explain these different outcomes. PMID- 12903849 TI - How reliable is the evidence for screening mammography? AB - Substantial reduction in breast cancer mortality has been proven in randomized trials of screening mammography. These trials have found a statistically significant benefit both for women ages 40-49 years and for women age 50 years and older at onset of screening. In fact, it is likely that the trials actually underestimate the benefit for an individual woman who is screened. Service screening programs have shown a 63% reduction in deaths from breast cancer among women who are screened. Virtually all major medical organizations now advise screening mammography for women ages 40 and older. Two recent studies that questioned the validity of results from screening mammography trials are themselves fatally flawed. PMID- 12903851 TI - Skin cancer chemoprevention: strategies to save our skin. AB - There are over 1 million cases of skin cancer diagnosed yearly in the United States. The majority of these are nonmelanoma (NMSCs) and are associated with chronic exposure to ultraviolet light (UV). Actinic keratosis (AK) has been identified as a precursor for SCC, but not for BCC. AKs are far more common than SCC, making them excellent targets for chemoprevention. Cancer chemoprevention can prevent or delay the occurrence of cancer in high-risk populations using dietary or chemical interventions. We have developed strategies that have rational mechanisms of action and demonstrate activity in preclinical models of skin cancer. Promising agents proceed to phase I-III trials in subjects at high risk of skin cancer. UV light induces molecular signaling pathways and results in specific genetic alterations (i.e., mutation of p53) that are likely critical to skin cancer development. UVB-induced changes serve as a basis for the development of novel agents. Targets include inhibition of polyamine or prostaglandin synthesis, specific retinoid receptors, and components of the Ras and MAP kinase signaling pathways. Agents under study include: epigallocatechin gallate (EGCG), a green tea catechin with antioxidant and sunscreen activity, as well as UVB signal transduction blocking activity; perillyl alcohol, a monoterpene derived from citrus peel that inhibits Ras farnesylation; difluoromethylornithine (DFMO), an inhibitor of ornithine decarboxylase and polyamines; retinoids that target retinoid X receptors and AP-1 activity; and nonsteroidal anti-inflammatory agents that inhibit cylooxygenase and prostaglandin synthesis. We performed a series of Phase I-II trials in subjects with multiple AK. For example, a phase II randomized trial of topical DFMO reduced AK number, suppressed polyamines, and reduced p53 protein. Our goal is to develop agents for use in combination and/or incorporation into sunscreens to improve chemoprevention efficacy and reduce skin cancer incidence. PMID- 12903853 TI - Chemoprevention of lung cancer: new directions. AB - The refractoriness of advanced lung cancer to current treatment modalities requires new approaches to reduce the public health burden associated with this disease. One strategy that is currently being tested is chemoprevention, which aims to prevent the development of cancer in populations that are at high risk for cancer due to a variety of genetic or environmental factors. The key to the success of this approach, however, requires the identification of appropriately targeted efficacious, non-toxic agents as well as the methodologies to efficiently test them. Given the lack of success of previous phase III definitive lung cancer chemoprevention trials, there is a need for smaller scale phase II trials with molecular, imaging, or histologic endpoints to demonstrate preliminary safety and efficacy. The identification of molecular pathways critical to lung carcinogenesis offers the opportunity to develop targeted therapies for prevention. Means of optimizing the risk/benefit ratio associated with treatment include regional drug delivery that minimizes systemic toxicities and combination therapies. Identification of the most appropriate cohorts, such as former smokers without ongoing DNA damage due to carcinogen exposure, may uncover benefits that are hidden in a mixed population. Equally important is the identification of appropriate study endpoints that are predictive of patient outcomes such as cancer incidence. Further understanding of lung cancer biology will be critical to the success of future clinical trials. PMID- 12903852 TI - Chemoprevention of nonmelanoma skin cancer: experience with a polyphenol from green tea. AB - Nonmelanoma skin cancer is extremely common and is increasing in incidence. It would be very useful to have forms of therapy that would prevent precancerous changes from going on to form cancer, or to reverse the precancerous changes. Epidemiologic evidence in humans, in vitro studies on human cells, and clinical experiments in animals have identified polyphenol compounds found in tea to be possibly useful in reducing the incidence of various cancers, including skin cancer. To examine the potential for a polyphenol from green tea, epigallocatechin gallate, to act as a chemopreventive agent for nonmelanoma skin cancer, a randomized, double-blind, placebo-controlled phase II clinical trial of topical epigallocatechin gallate in the prevention of nonmelanoma skin cancer was performed. PMID- 12903854 TI - Key issues in lung cancer chemoprevention trials of new agents. AB - Lung cancer is a major health problem world-wide. Former heavy smokers retain a significant risk for lung cancer after smoking cessation. With a large population of current and former smokers at risk, an alternative cancer control strategy such as chemoprevention needs to be developed to reduce lung cancer mortality especially for smokers who have followed medical advice to give up smoking. Currently, there is no agent that has been shown to be effective in preventing lung cancer. Key issues that need to be addressed in phase II trials of promising chemopreventive agents include selection of high-risk subjects, potential variation in response due to differences in gender and smoking history as well as the choice of surrogate endpoint biomarkers. Sputum biomarkers such as image analysis of sputum cells and detection of aberrant methylation hold promise in identifying those at highest risk for chemopreventive intervention. Autofluorescence bronchoscopy is an effective method to localize dysplastic lesions to evaluate the efficacy of new chemopreventive agents. Novel imaging methods such as confocal micro-endoscopy and spiral CT-directed endoscopic biopsies are under development to evaluate the response of chemopreventive agents on peripheral pre-neoplastic lesions in small airways. PMID- 12903855 TI - Prevention of prostate cancer. AB - Prostate cancer is an ideal candidate for chemoprevention because of its high prevalence, long latency time, hormone dependency, precursor lesions, and its unique serum marker, PSA. Chemoprevention is the administration of drugs or other agents which aim to prevent the induction or inhibit/delay cancer progression. Large-scale studies favor environmental rather than genetic factors as key determinants of prostate cancer development. Among these environmental factors, nutrition certainly has a leading role. Numerous basic science studies but also clinical studies indicate that dietary compounds or diet modifications may ultimately play a major role in prostate cancer promotion and inhibition. Definitive proofs are often difficult because of methodological problems and complex triggering cascades. New pharmaceutical drugs with minimal toxicity are also currently evaluated. PMID- 12903856 TI - Clinical models for testing chemopreventative agents in prostate cancer and overview of SELECT: the Selenium and Vitamin E Cancer Prevention Trial. AB - Target populations for chemoprevention trials should include those at higher than average risk for the development of prostate cancer as defined by explicit epidemiologic and genetic criteria. Such populations include a "primary prevention" group without histologic or clinical evidence of cancer, and several clinical models of "secondary prevention," including those with clinically evident disease prior to definitive therapy and those at high risk of recurrence after therapy based on histology and/or biochemical status. Each risk group and clinical model has potential advantages and disadvantages, and the mechanisms which underlie disease development and progression in each group may be unique. These observations give rise to many potential clinical trials of specific agents. These trials should also include collection of data on potentially confounding influences on disease development and progression. Preclinical, epidemiologic, and Phase II data suggest that both selenium and vitamin E have potential efficacy in prostate cancer prevention. The experience of the Prostate Cancer Prevention Trial (PCPT) demonstrates the interest and dedication of healthy men to long-term studies of cancer prevention. SELECT, the Selenium and Vitamin E Cancer Prevention Trial, is an intergroup phase III, randomized, double blind, placebo-controlled, population-based clinical trial designed to test the efficacy of selenium and vitamin E alone and in combination in the prevention of prostate cancer which builds on secondary analyses of large-scale chemoprevention trials for other cancers and the lessons of PCPT. PMID- 12903857 TI - Problems with prostate-specific antigen screening: a critical review. AB - The effect of population screening with regard to reduction of prostate cancer specific mortality and quality of life issues is not yet clear. Several national and international prospective studies are currently being conducted to answer these important questions. They include the trials in the Federal State of Tyrol, Austria and in the Quebec City area, Canada, as well as the Prostate, Lung, Colorectal and Ovarian (PLCO) trial in the United States and the European Randomized Study of Screening for Prostate Cancer (ERSPC). In the meantime, individual case finding (opportunistic screening) is recommended for men with a life-expectancy of at least 10 years. PMID- 12903858 TI - Genetic predisposition as a basis for chemoprevention, surgical and other interventions in colorectal cancer. AB - Strategies of cancer prevention are generally developed with the population at large in mind. However, special attention is warranted for those persons with rare genetic traits associated with a greatly elevated risk of developing colorectal cancer (CRC) and some other malignancies: Orphan diseases demand Orphan preventive measures! Recent advances in modern genetics have enhanced our understanding of several genes and the specific germ-line mutations responsible for colorectal carcinogenesis. A number of features provide evidence for a genetic predisposition to CRC. These include typical clinical and histological features of a particular syndrome, a familial aggregation of CRC and associated malignancies, young age at onset of CRC, occurrence of multiple neoplasias and/or unusual localisation of the tumour (e.g., right side of the colon). In hereditary colorectal cancer, genetic testing can easily be demonstrated as cost-effective. PMID- 12903859 TI - Fecal occult blood screening--trial evidence, practice and beyond. AB - Colorectal cancer is a leading cause of morbidity and mortality in Western countries. Primary prevention and early detection of this malignancy has been shown in multiple research set-ups, and using different modalities, to be effective in reducing the incidence and mortality rates of this disease. Three randomized controlled trials have been conducted over the last 30 years to evaluate the efficacy of periodic screening with fecal occult blood tests (FOBT). These studies have consistently demonstrated mortality reduction with biennial as well as annual testing after 8-18 years of follow-up. A significant primary prevention effect through reduction of colorectal cancer incidence was also reported. The results of the Israeli population-based screening program using Hemoccult Sensa show that it is possible to achieve a high detection rate in a well-organized community set-up and, in addition, also a shift in tumor stage towards smaller tumors, a low positivity rate, and an acceptable false positivity rate. FOBT is cheap and performs very well in cost effectiveness analyses evaluating the cost of detecting one cancer. This is quite a rare situation where mortality reduction can be achieved with a simple rather than a sophisticated technology. Research to further enhance the cancer detection capabilities of FOBT through the incorporation of molecular testing of the stool for tumor genes is underway. Multiple policymakers are already recommending this procedure for routine screening of average risk population. Taking their recommendations into action is of major importance. PMID- 12903860 TI - Is FOB screening really the answer for lowering mortality in colorectal cancer? AB - In the three trials that tested screening with biennial fecal occult blood test (FOBT), follow-up of control patients for colorectal cancer (CRC) differed: in the Minnesota (United States) trial, the follow-up was equivalent to patients in the intervention groups, while in the Nottingham (United Kingdom) and Funen (Denmark) trials, control patients just received usual care. In the two latter trials, mortality from colorectal cancer was lower in subjects with interval colorectal cancer than in control subjects, while in the Minnesota trial, survival was equivalent in patients with interval CRC and in control patients. We examined whether better disease awareness of subjects allocated to the intervention group contributed to changes in colorectal cancer mortality observed in the FOBT trials. In the Nottingham and Funen trials, we evaluated the amount of colorectal cancer mortality reduction attributable to better survival of subjects in whom an interval colorectal cancer developed. In the Minnesota trial, we examined whether earlier detection of colorectal cancer in control subjects could explain the small (6%) reduction in colorectal cancer mortality observed with biennial FOBT. In the Nottingham and Funen trials, about one-quarter of the reduction in colorectal cancer mortality could be attributed to better awareness of patients with interval colorectal cancer. After correction for the effects of disease awareness, the absolute reduction in colorectal cancer mortality due to FOBT itself was 12% instead of 16%, and was no longer statistically significant (P>0.05). The results from biennial FOBT in the Minnesota trial published in 1993 would probably have been similar to those obtained in the Nottingham and Funen trials if disease awareness had not influenced the stage at diagnosis of colorectal cancers found in the control group. Better awareness of colorectal cancer contributes to the reduction of colorectal cancer mortality and should be encouraged. Because of a study design effect, the decrease in colorectal cancer mortality attributable to the FOBT itself is about 25% lower than that reported in the Nottingham and Funen trials. Therefore, recommending general population screening with biennial FOBT is still an open question. PMID- 12903861 TI - Effects of fish oil on lymphocyte proliferation, cytokine production and intracellular signalling in weanling pigs. AB - It has been widely documented that fish oil attenuates inflammatory responses partially via down-regulation of T-lymphocyte function. To determine the anti inflammatory role of fish oil in weanling pigs, we investigated the effects of fish oil and its functional constituents on peripheral blood lymphocyte proliferation, cytokine production and subsequent intracellular signalling in inflammatory-challenged weanling pig and in in vitro cultured lymphocytes. Fish oil (7%) or corn oil (7%) was supplemented to 72 crossbred pig (7.6 +/- 0.3 kg BW and 28 +/- 3 days of age) in a 2 x 2 factorial experiment that included an Eacherichia coil lipopolysaccharide (LPS) challenge (challenged or not challenged). On day 14 and 28 of the experiment, 200 microg/kg BW of LPS or an equivalent amount of sterile saline was administered to the pigs by intraperitoneal injection. Blood samples were collected on days 15 and 29 to determine peripheral blood lymphocyte proliferation, interleukin-1beta (IL-1beta) and interleukin-2 (IL-2) production. The results showed that inflammatory challenge decreased average daily gain (P < 0.05) and average daily feed intake (P < 0.05) during days 15-28. Fish oil supplementation had no effect on growth performance. Inflammatory challenge increased lymphocyte proliferative response to concanavalin A (Con A) (P < 0.05) following each challenge. Fish oil tended to suppress (P < 0.1) the proliferation following the first challenge. Similarly, fish oil tended to reduce IL-1beta production (P < 0.1) following the second challenge and IL-2 (P < 0.1) production following the first challenge in both challenged and unchallenged pigs compared with corn oil. In parallel in vitro experiments, peripheral blood lymphocytes of weanling pigs were incubated with various concentrations of eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA) or linoleic acid (LA) (0, 20, 40, 60, 80, 100 microg/ml). EPA, DHA and high levels of LA predominantly suppressed IL-1beta (P < 0.05), IL-2 (P < 0.05) production and subsequent lymphocyte proliferation (P < 0.05). Low levels of LA increased (P < 0.05) IL-2 production. Compared with LA, EPA resulted in a stronger inhibition of lymphocyte proliferation (P < 0.05) and IL-2 (P < 0.01), and DHA resulted in a stronger inhibition of IL-1beta (P < 0.05) and IL-2 (P < 0.01). To elucidate the mechanism(s) by which fish oil and its functional constituents suppressed lymphocyte function, the kinetics of intracellular [Ca2+]i and protein kinase C activity were determined in in vitro experiments. EPA, DHA and LA exerted very similar dose-dependent stimulatory effects on intracellular Ca2+. EPA and DHA inhibited protein kinase C activity (P < 0.05), while LA had no significant effect (P > 0.05). These results suggest that fish oil and its functional constituents (EPA and DHA) exerted an anti-inflammatory effect by down-regulation of lymphocyte activation in weanling pigs, possibly by manipulation of intracellular signalling. PMID- 12903862 TI - Prediction of digestibility and energy concentration of winter pasture forage and herbage of low-input grassland--a comparison of methods. AB - Regarding the estimation of the energy concentration or digestibility of herb dominated forage and plant samples from winter pastures, it could be expected that the estimation is only reliable when in vitro methods with rumen fluid as inoculum (= gas production techniques) are used. For the verification of this thesis based on logical reflections, an in vitro-method with rumen fluid added as inoculum, as well as chemical, and enzymatic methods were applied under consideration of existing estimating functions. As a possible reason for the observed divergence of the methods, effects of fungal infections or, respectively, secondary compounds in herbs are discussed. At the present state of knowledge, it is adequate to estimate the energy concentration in vitro by gas tests, as far as fattening types like suckler cows and beef cattle are concerned, maybe in contrast to the forage evaluation for dairy cows. PMID- 12903863 TI - Evaluation and interpretation of results for three cross-over designs. AB - In cross-over designs, individual sequences of treatments are applied to the animals. Within such designs it is possible that every treatment could modify the effect of the subsequent treatment applied to the same animal. We compared three cross-over designs each with three treatments, three periods, and two blocks. This comparison was done with respect to the variance of the estimations of the effects and its biases caused by the interactions between the treatment and the carry over effect of the foregoing treatment. Moreover, different methods of estimating variance components and calculating the degrees of freedom were compared by means of simulation. If the animal variance component is small, then the bias of the REML estimator of the variance components is greater than one of the widespread ANOVA-estimator called 'TYPE3'. But nevertheless, the mean squared error of this estimation is smaller in the case of REML in comparison to ANOVA. Therefore, the REML method should be preferred. For calculating the degrees of freedom, the Kenward-Roger method should be used. After applying this method, the true significance level is almost equal to its required value, but if the Satterthwaite method is used, the true significance level will be too high. If the interaction (treatment x carry over) is ignored in the model although it exists, the standard error of the treatment effect estimation is too great, and, therefore, the true significance level is too small. The methods which have been evaluated are available in the SAS-procedure MIXED (SAS Institute, 1999a). To assist the investigation of cross-over designs by using this software, we developed programs for data management and data analysis. These programs are available from the first author. PMID- 12903864 TI - Effects of dietary vitamin K levels on bone quality in broilers. AB - This study was conducted to evaluate the effects of dietary vitamin K (menadione) on bone quality in cage-raised broilers. Three hundred and sixty male broilers were randomly allotted to one of six treatments, with six replicate pens per treatment and 10 chicks per pen. Broilers were fed one of six diets including a control diet or the control diet plus graded levels of vitamin K (0.5 mg/kg, 2 mg/kg, 8 mg/kg, 32 mg/kg and 128 mg/kg). Water and feed were provided ad libitum during the 7-week experimental period. Results indicated that vitamin K supplementation of broilers diets significantly effected bone quality and feed efficiency. The treatment containing vitamin K at 8 mg/kg improved growth performance (during weeks 6-7) and bone quality (during weeks 0-3). In our study, hydroxyapatite binding capacity of serum osteocalcin (during weeks 0-3), bone breaking strength, bone flexibility, bone ash weight increased linearly (P < 0.05) and bone mineral density, bone mineral content increased quadratically (P < 0.05) with increasing supplementation of vitamin K. In conclusion, to gain optimum bone quality and broiler performance, our studies suggest that the concentration of vitamin K in broilers diets should be 8 mg/kg, 2 mg/kg, and 2 mg/kg, for the starter, grower and finisher phases, respectively. Furthermore, it was shown that the starter period is an important phase for improving bone quality. In addition, this study validated the mechanism of vitamin K effects on bone quality. Vitamin K boosts the carboxylation of osteocalcin and decreases the concentration of serum under-carboxylated osteocalcin enhancing hydroxyapatite binding capacity of serum osteocalcin and improving bone quality. PMID- 12903865 TI - In vivo antioxidant properties of vitamin E and chromium in cold-stressed Japanese quails. AB - An experiment was conducted to determine if vitamin E (alpha-tocopherol-acetate) and chromium (chromium picolinate, Cr Pic) supplementation attenuate the negative effects of cold stress on egg production, egg quality, serum metabolites, and antioxidant status in Japanese quails (Corurnix coturnix japonica). One hundred and fifty laying Japanese quails (50-day-old) were divided into five groups, 30 birds per group. The laying quails kept at 6 degrees C for 12 h/d (08.00 p.m. to 08.00 a.m.) were fed either a basal diet (low temperature-basal diet, CS group) or the basal diet supplemented with either 400 microg of Cr/kg of diet (Cr group), 250 mg of alpha-tocopherol-acetate per kg of diet (Vit. E group) or 400 microg of Cr plus 250 mg of alpha-tocopherol-acetate per kg of diet (Vit. E + Cr group) while quails kept at 18 degrees C were fed a basal diet (thermo-neutral basal diet, TN group). Performance and egg quality were significantly reduced in CS group compared with TN group. Supplemental chromium and vitamin E significantly increased live weight change, egg production, and improved feed efficiency in cold-stressed laying hens compared with the group fed the basal diet at 6 degrees C. Egg production and egg weight were also greater (P < 0.05) in each supplemental group compared with the CS group. However, a combination of vitamin E and chromium, rather than each separately, provided the greatest performance. Supplemental vitamin E and chromium also increased serum vitamin C and E but, decreased malondialdehyde (MDA) concentrations (P < 0.05); the combination of vitamin E and chromium resulted in the highest levels of serum vitamin C and E within the cold-stressed quails. Results of the present study indicate that combined antioxidant supplements increased performance, egg quality and serum antioxidant levels while lowering MDA in cold-stressed quails. PMID- 12903866 TI - Development of the digestive tract in the ostrich (Struthio camelus). AB - A study was conducted to investigate changes in the development of the gastrointestinal tract (GIT) in relation to body growth of growing ostriches. There was an 11-fold increase (P < 0.001) in body weight between 3 and 72 days of age. The relative (to body weight) weight of the proventriculus/gizzard, caeca and colon also increased (P < 0.001) with age. The relative weight of the small intestine peaked at 41 days of age and then tended to decline (P < 0.05) subsequently. The relative weight of the pancreas peaked at 27 days of age and remained fairly stable thereafter. The activities of chymotrypsin and lipase declined (P < 0.001) with age between 3 and 72 days. At 3 days of age, the protein content of the duodenal mucosal homogenate was higher (P < 0.001) than that of the jejunum or ileum, but at all subsequent periods the jejunal protein content was the highest. The protein content of the intestinal brush-border membrane was higher (P < 0.001) at the jejunum than at the duodenum or ileum. The specific activity of maltase declined (P < 0.001) with age in all three regions, most especially between 3 and 27 days of age. The activity of alkaline phosphatase (AP) at 41 and 55 days of age was higher (P < 0.001) in the duodenum than in the jejunum or ileum. The activity of AP fluctuated with age in the duodenum but there was a more defined decline (P < 0.001) with age in the jejunum and ileum. The relative protein content of the liver increased (P < 0.001) with age, with two peaks at 27 and 55 days of age. Arginase activity was not detected in the liver of 3-day old chicks and was not significantly affected by age between 27 and 72 days of age. The pattern of development observed is similar to that in growing poultry. There is, however, a need for evaluation at closer intervals in early life as well as an in-depth assessment of the morphometry of the intestinal mucosa. PMID- 12903867 TI - The effect of a probiotic Enterococcus faecium product in diets of healthy dogs on bacteriological counts of Salmonella spp., Campylobacter spp. and Clostridium spp. in faeces. AB - An experiment was conducted to determine the effects of a probiotic on selected faecal bacteria of healthy dogs under different feed and environmental conditions. For the study 12 dogs kept in households were used for an 18-day supplementation with a patented commercially available strain of E. faecium NCIB 10415 (Enteroferm). In order to minimize losses the probiotic product was orally applicated once a day before meals at a dose of 2 g per dog (9.2 x 10(9) CFU). The faeces were collected before the beginning of the supplementation and at the end of the 18-day application period. In order to exclude contamination, all faeces were taken rectally. Before and at the end of the experimental period total Salmonella spp., Campylobacter spp. and Clostridium spp. counts were determined in fresh faeces using selective media. It was demonstrated that the 18 day application of the probiotic E. faecium product induced modifications on the gastrointestinal microflora in all dogs. While Salmonella spp. and Campylobacter spp. counts were in majority of the dogs higher than before the application. Clostridium spp. counts were significantly reduced in 10 of 12 dogs. According to the guidelines for the evaluation of the efficiency of microorganisms in dogs a relevant efficacy effect was supported by this data. However, a beneficial effect of the probiotic product on healthy dogs remains questionable. PMID- 12903868 TI - Pulmonary hypertension in infancy and childhood. AB - In this review, we discuss current concepts in the pathogenesis and management of pulmonary hypertension affecting infants and children, with special focus on left to-right shunting, bronchopulmonary dysplasia, and primary pulmonary hypertension. In patients of these ages, functional aspects, such as an imbalance between vasoconstricting and vasodilating mechanisms, and morphological alterations of the vessel wall, contribute to the pulmonary hypertension. In the past decades, strategies have emerged for treatment that are targeted at the pathophysiological basis. Thus, in patients with left-to-right shunting and pulmonary hypertension after intra-cardiac repair, treatment with nitric oxide has been introduced effectively, while treatment with prostanoids, such as iloprost, is under investigation. In patients with pulmonary hypertension and bronchopulmonary dysplasia, therapeutic strategies focus on the underlying chronic lung disease and use of vasodilators. The pathogenesis of primary pulmonary hypertension in children remains as yet unclear, although treatment with prostanoids has proven effectively to improve the long-term prognosis. PMID- 12903869 TI - The clinical implications of the systemic inflammatory reaction related to cardiac operations in children. PMID- 12903870 TI - An echocardiographic study of tetralogy of Fallot in the fetus and infant. AB - OBJECTIVE: To document the echocardiographic features of tetralogy of Fallot during fetal and postnatal life. Correlation of echocardiographic findings with the requirement for early intervention prior to definitive repair. DESIGN: Retrospective observational study. SETTING: A tertiary fetal cardiology unit. PATIENTS: Fetuses with a diagnosis of tetralogy of Fallot identified from a prospective database between 1 January 1999 and 31 October 2002. MAIN MEASURES OF OUTCOME: Growth of aorta and pulmonary trunk during fetal and postnatal life. Doppler assessment of the great arteries both prenatally and postnatally. Clinical outcome to definitive repair. RESULTS: We identified 25 fetuses with tetralogy of Fallot, 23 having a pulmonary valvar diameter below the normal range at some point during gestation. The ratio of the diameter of the aortic to the pulmonary valve was abnormal in all cases. The pulmonary arterial Doppler velocity was within the normal range in six fetuses at presentation,and elevated in the remainder. In two fetuses, the right ventricular outflow tract was patent during fetal life, but had become atretic at birth. Both of these fetuses had reversal of flow in the arterial duct at presentation during fetal life. In 2 fetuses in whom we showed poor growth of the pulmonary trunk in late gestation, it was necessary to intervene early. The Doppler velocity across the pulmonary valve during fetal life did not differentiate between babies who required early intervention and those who were repaired electively. There was a marked increase in pulmonary arterial Doppler velocity following birth, which became more elevated with age. Of 18 liveborn infants, 17 have survived, with 2 having balloon dilation of the right ventricular outflow tract, and 3 insertion of a Blalock-Taussig shunt prior to definitive repair. CONCLUSIONS: In tetralogy of Fallot, features of pulmonary valvar hypoplasia and obstruction are evident during fetal life. Progression of obstruction in the right ventricular outflow tract was observed during fetal life as well as postnatally. Reversal of flow in the arterial duct, and failure of growth of the pulmonary trunk, predicted the need for early surgery to maintain pulmonary blood flow. Parents should be counselled about the possibility of emergency intervention being required after birth. Affected fetuses should be delivered at units with experience of managing the cyanosed newborn. PMID- 12903871 TI - Cardiac troponin-I in the serum of infants of diabetic mothers. AB - A transient form of hypertrophic cardiomyopathy has been previously described in infants of diabetic mothers. When it occurs, it is generally benign. The purpose of our study was to establish the extent of injury to the cardiomyocytes in symptomatic and asymptomatic patients with and without hypertrophic cardiomyopathy. Thus, we compared 35 consecutive patients to 20 healthy controls, establishing the significance, if any, of differences in cardiac troponin-I and creatine kinase, including its MB-fraction, and seeking to establish the value of these parameters in the diagnosis of cardiac injury. We also determined to levels of glucose and insulin in the serum, and took note of electrocardiographic and echocardiographic investigations. Values were determined at the 1st and 7th days after admission in the patients, while parameters were measured in the control group only on the first day. We found that the levels of cardiac troponin-I in the serum, known to be a marker for cardiac injury, were significantly elevated in symptomatic patients with life-threatening respiratory or haemodynamic distress. We speculate that transient ventricular hypertrophy is neither the cause nor the consequence of damage to the cardiomyocytes. It would be interesting, nonetheless, to determine the relationship, if any, between cardiomyocytic damage and clinical outcome. PMID- 12903872 TI - Optimal timing of the Ross procedure in the management of chronic aortic incompetence in the young. AB - The appropriate timing of intervention in patients with chronic aortic incompetence allows recovery of ventricular function. We sought to determine the optimal timing of the Ross procedure for chronic aortic incompetence in young patients. We retrospectively analysed case notes, and measured pre- and postoperative echocardiographic indexes of left ventricular function, in patients who had undergone the Ross procedure for chronic aortic incompetence. METHODS AND RESULTS: We found 21 patients with preoperative and postoperative data suitable for analysis. Their age at operation ranged from 5.6 to 26 years, with a median of 13.8 years, and the duration of follow-up was from 0.5 to 6.8 years, with a median of 2.4 years. The preoperative left ventricular end-diastolic dimension was converted to a z-score, and this was used as a threshold to divide the population. Using the threshold of a preoperative left ventricular z-score of more than 3 to divide the population did not show any difference in postoperative parameters of left ventricular function. Significant differences were found postoperatively, however, in both the left ventricular z-score and the ratio of left ventricular end-diastolic radius to posterior wall thickness in diastole, with a cutoff preoperative threshold z-score greater than 4. CONCLUSION: The increase in the ratio of left ventricular end-diastolic radius to the thickness of the posterior wall in diastole would suggest that there is disruption of left ventricular short axis architecture and myocardial contractile function when intervention is postponed. The significantly larger left ventricular dimension at end-diastole, despite the reduction in volume loading post surgery, may also demonstrate irreversible structural changes. Our data would suggest that recovery of left ventricular function is less likely when the left ventricular z-score has reached the value of 4, and that, ideally, intervention should be performed when the z-score approaches or exceeds 3. PMID- 12903873 TI - Electrocardiographic changes in patients with cardiac rhabdomyomas associated with tuberous sclerosis. AB - BACKGROUND: Cardiac rhabdomyomas associated with tuberous sclerosis induce various abnormalities in the electrocardiogram. Electrocardiographic evidence of ventricular hypertrophy may appear if the tumour is electrically active. To our knowledge, electrocardiographic evidence of ventricular hypertrophy has been reported only in association with congestive heart failure. Follow-up studies of changes in electrocardiographic findings are also lacking. METHODS: We studied 21 consecutive patients with cardiac rhabdomyoma associated with tuberous sclerosis, 10 males and 11 females, aged from the date of birth to 9 years at diagnosis. The mean period of follow-up was 53 months. None of the patients developed congestive heart failure. We evaluated the electrocardiographic changes during the follow up, and their association with echocardiographic findings. RESULTS: Of the 21 patients, 12 showed one or more abnormalities on the electrocardiogram at presentation, with five demonstrating right or left ventricular hypertrophy. In all of these five cases, the tumours were mainly located in the respective ventricular cavity. In one patient with a giant tumour expanding exteriorly, there was marked left ventricular hypertrophy on the electrocardiogram. Followup studies showed spontaneous regression of the tumours in 12 of 19 patients, with abnormalities still present in only 7 patients. A gradual disappearance of left ventricular hypertrophy as seen on the electrocardiogram was noted in the patient with marked left ventricular hypertrophy at presentation in parallel with regression of the tumour. CONCLUSIONS: The presence of cardiac rhabdomyomas in patients with tuberous sclerosis might explain the ventricular hypertrophy seen on the electrocardiogram through its electrically active tissue without ventricular pressure overload or ventricular enlargement, although pre-excitation might affect the amplitude of the QRS complex. Even in cases with large tumours, nonetheless, the electric potential might- not alter the surface electrocardiogram if the direction of growth of the tumour is towards the ventricular cavity. In many cases, electrocardiographic abnormalities tend to disappear, concomitant with regression of the tumours. PMID- 12903874 TI - Thoracic sequels after thoracotomies in children with congenital cardiac disease. AB - The standard surgical approach for closed heart procedures in small infants and children is to use a posterolateral thoracotomy incision, which results in the division of the latissimus dorsi and serratus anterior muscles. The aim of our study was to determine the frequency and type of musculoskeletal deformities in children undergoing surgery with this approach for congenital cardiac disease. We included 49 children, 28 boys and 21 girls, in the study. Their mean age was 10.2 +/- 4.8 years, the mean age at the time of surgery was 3.8 +/- 4.0 years, and they were evaluated at an average of 6 years after the thoracotomy. Of the patients, 94% had various musculoskeletal deformities. Scoliosis was observed in 15 patients (31%) but only in two patients did the curves exceed 25 degrees. Of these patients, three-fifths had aortic coarctation. Elevation of the shoulder was seen in 61%, winged scapula in 77%; while 14% had asymmetry of the thoracic wall due to the atrophy of the serratus anterior muscle. Deformity of the thoracic cage was observed in 18%; and 63% had asymmetry of the nipples. Thus, we found that musculoskeletal deformities are frequent after thoracotomies in children with congenital cardiac disease. Patients who have undergone such procedures for cardiac or noncardiac surgery should be followed until their skeletal maturation is complete. Techniques sparing the serratus anterior and latissimus dorsi muscles should be preferred. These adverse effects of thoracotomy may be another reason for using interventional procedures in these cases. PMID- 12903875 TI - Increased levels of brain and atrial natriuretic peptides after the first palliative operation, but not after a bidirectional glenn anastomosis, in children with functionally univentricular hearts. AB - We evaluated the concentrations of the brain and atrial natriuretic peptides in the plasma as markers of ventricular function and volume load in children with functionally univentricular hearts. We studied 7 children aged from 0.5 to 0.7 years with functionally univentricular hearts who had undergone a first palliative operation, and 10 children aged from 1.8 to 3.7 years who had undergone a bidirectional Glenn anastomosis at ages ranging from 0.4 to 1.0 year. As a control group, we studied 14 children without heart defects aged from 0.1 to 4.5 years. Levels of the brain natriuretic peptide were measured at 8.3 to 122 ng/l, with a mean of 52.8 ng/l, after the first palliative operation, compared to 0 to 16 ng/l, with a mean of 7.3 ng/l, after a bidirectional Glenn anastomosis, and 0 to 13.8 ng/l, with a mean of 5.9 ng/l, in the children serving as controls. Corresponding values for atrial natriuretic peptide were 17 to 203 ng/l, with a mean of 103 ng/l, after the first palliative operation, compared to 16 to 54 ng/l, with a mean of 29 ng/l, after the bidirectional Glenn anastomosis, and 12 to 52 ng/l, with a mean of 32 ng/l in the controls. Echocardiography showed that all the children with functionally univentricular hearts had normal ventricular function. Blood presssure, pulmonary arterial pressure, and arterial saturations of oxygen did not differ between the groups. We conclude, that in children with functionally univentricular hearts, the volume overload imposed on the heart after the first palliative operation is associated with increased production of brain and atrial natriuretic peptides, while after ventricular unloading, levels of the natriuretic peptides return to control values. PMID- 12903876 TI - Cardiac catheterization in children as outpatients: potential, eligibility, safety and costs. AB - BACKGROUND: Outcomes and costs of inpatient versus outpatient pediatric cardiac catheterization have not been extensively evaluated. METHODS: For a cost consequence analysis, we reviewed the medical records and cost data in a Swiss pediatric hospital. We compared outcomes and costs of observed inpatient management versus hypothetical planned same-day discharge for patients meeting the outpatient catheterization criterions for an American pediatric hospital. RESULTS: Among 346 catheterization admissions occurring from January, 1998 through December, 1999, 179 met the American criterions for outpatient catheterization. Complications observed, and/or nursing interventions begun within 5 hours of catheterization, might have required overnight observation in 41 of the 179 admissions (22.9%). The remaining 138 patients were stable at five hours, and presumably could have been discharged the day of the procedure. Routine pre-discharge imaging detected significant complications following three interventional procedures. Postulated costs from the perspective of the provider, counting hospital and physician expenditure were calculated for the Swiss franc in 2000 at a rate of 1.69 francs for each American dollar, averaged 10,946 francs per inpatient, versus 9790 francs for outpatient treatment (p < 0.001 by paired t test). Estimated revenue deficits, calculated as costs minus reimbursement, averaged 8565 francs per inpatient versus 1756 francs per patient treated as an outpatient. CONCLUSIONS: Half the patients being catheterized in the Swiss hospital met the external criterions for attempted outpatient catheterization. Most might have been safely discharged on the same day, with modest savings in costs, and reduced deficits in terms of revenue. Routine predischarge imaging may be more important than overnight observation. Outpatient catheterization merits prospective evaluation in Switzerland. PMID- 12903877 TI - Percutaneous transluminal coronary angioplasty for anastomotic stenosis after coronary arterial bypass grafting in Kawasaki disease. AB - OBJECTIVES: We evaluated the efficacy of percutaneous transluminal coronary angioplasty for anastomotic stenosis after coronary arterial bypass grafting using the internal thoracic artery in patients with coronary arterial lesions due to Kawasaki disease. SUBJECTS AND METHODS: From July 1997 to April 2000, four boys and one girl underwent percutaneous transluminal coronary angioplasty for 6 anastomotic lesions following coronary arterial bypass grafting using the left or right internal thoracic artery. Progressive severe stenosis of the grafts in the follow-up angiograms after grafting, and evidence of ischemia, were regarded as indications for percutaneous transluminal coronary angioplasty. Age at coronary angioplasty ranged from 4.2 to 16.7 years, with a median of 6.9 years, while the interval from operation ranged from 0.3 to 3.0 years, with a median of 1.1 years. The diameter of the balloon catheter employed varied from 1.5 to 2.5 mm, and the pressure of inflation ranged from 8 to 16 atmospheres. RESULTS: The degree of stenosis decreased from 63 to 99%, with a median of 88%, to 0 to 40%, with a median of 17% immediately after angioplasty. A follow-up angiogram either 3 months or 1 year later revealed no restenosis in any patient. CONCLUSION: Percutaneous transluminal coronary angioplasty is a feasible and useful procedure for treating anastomotic stenosis following coronary arterial bypass grafting using the internal thoracic artery in patients with coronary arterial lesions due to Kawasaki disease. PMID- 12903879 TI - Arteriovenous fistula of the internal thoracic artery. PMID- 12903878 TI - Echocardiographic considerations during deployment of the Helex Septal Occluder for closure of atrial septal defects. AB - The Helex Septal Occluder is a new device used to close atrial septal defects via interventional catheterization. In order to study the role of echocardiography during its use, and to describe the morphologic variants of defects suitable for closure with this occluder, we evaluated all patients undergoing intended closure of an atrial septal defect with the Helex occluder. A combination of transthoracic, transesophageal, three-dimensional, and intracardiac echocardiography were used before, during, and after the procedure to characterize anatomy, assess candidacy for closure, guide the device during its deployment, and evaluate results. Among the 60 candidates included in the study, 11 were excluded because of transesophageal echocardiographic and/or catheterization data obtained in the laboratory. Attempts at closure were successful in 46 patients, and unsuccessful in 3. We successfully treated four types of defects. These were defects positioned centrally within the oval fossa with appreciable rims along the entire circumference of the defect, defects with deficient or absent segments of the rim, defects with aneurysm of the primary atrial septum, and defects with multiple fenestrations. Follow-up transthoracic echocardiograms taken at a median of 7 months demonstrated no residual defects in 21, trivial residual defects in 17, and small residual defects in 8 patients. In 20 patients, three-dimensional reconstructions were used to characterize the morphology of the defect and the position of the device. Because transesophageal echocardiography was often limited by acoustic interference from the device, intracardiac echocardiography was utilized in 3 cases to overcome this limitation. PMID- 12903880 TI - Aortic stenosis after uncomplicated surgical repair of tetralogy of Fallot. AB - Tetralogy of Fallot is only rarely associated with aortic valvar disease. We present a child who had uncomplicated repair of tetralogy of Fallot at 16 months of age, and who developed mild aortic stenosis three years later, with mild to moderate aortic incompetence being seen two years subsequent to that event. No aortic valvar disease had been noted prior to surgery. No intervention is planned at this stage. PMID- 12903881 TI - Hoarseness after pulmonary arterial stenting and occlusion of the arterial duct. AB - We report a 12-year-old girl who had multiple congenital cardiac lesions, specifically an arterial duct, left pulmonary arterial stenosis, an atrial septal defect in the oval fossa, and mild Ebstein's malformation of the tricuspid valve. Therapeutic transcatheter intervention was performed to stent the left pulmonary artery, occlude the arterial duct with a coil, and place a device to close the atrial septal defect. Subsequent to the catheterization, she complained of hoarseness, which was shown to be due to entrapment of the left recurrent laryngeal nerve between the coil used to close the arterial duct and the stent placed in the left pulmonary artery. Laryngoscopy confirmed paralysis of the recurrent laryngeal nerve. PMID- 12903882 TI - Amiodarone-induced 2 to 1 atrioventricular block in association with prolongation of the QT interval. AB - The potential for development of 2 to 1 atrioventricular conduction in children with prolongation of the QT interval has been previously reported secondary to electrolytic disturbances. We report here a child who developed 2 to 1 atrioventricular conduction with prolongation of the QT interval following treatment with amiodarone for refractory supraventricular tachycardia. We highlight the importance of electrocardiographic monitoring to assess for those at risk of amiodarone toxicity, which may be manifested by prolongation of the QT interval and the simultaneous loss of atrioventricular conduction, and of equal importance the need for prompt conversion to an alternative anti-arrhythmic agent. PMID- 12903883 TI - Transcatheter reconstruction of the right heart. AB - We report the interventional procedures performed on a 12-year-old child with obstruction of the right ventricular outflow tract, pulmonary valvar insufficiency, pulmonary arterial stenosis, and an atrial septal defect. A staged repair of all anomalies was performed successfully using transcatheter techniques. PMID- 12903884 TI - Diffuse varices of the right pulmonary veins. AB - We describe a patient with diffuse varices of the right pulmonary veins in association with patency of the arterial duct and an atrioventricular septal defect with separate right and left atrioventricular valvar orifices. PMID- 12903885 TI - Arginine supplementation enhances peritoneal macrophage phagocytic activity in rats with gut-derived sepsis. AB - BACKGROUND: Previous reports have shown that arginine (Arg) enhances phagocytic activity of macrophages and is required for macrophage-mediated toxicity toward tumor cells. Few studies have addressed the importance of Arg supplementation on macrophage and neutrophil function after infection and sepsis. This study examined the effect of Arg-supplemented diets before and Arg-enriched total parenteral nutrition (TPN) after sepsis or both on the phagocytic activity of peritoneal macrophages and blood polymorphonuclear cells in rats with gut-derived sepsis. METHODS: Male Wistar rats were assigned to 4 groups. Groups 1 and 2 were fed a semipurified diet, while groups 3 and 4 had part of the casein replaced with 2% of total calories as Arg. After the experimental diets were administered for 10 days, sepsis was induced by cecal ligation and puncture (CLP); at the same time, an internal jugular vein was cannulated. All rats were maintained on TPN for 3 days. Groups 1 and 3 were infused with conventional TPN, while groups 2 and 4 were supplemented with Arg, replacing 10% of total amino acids in the TPN solution. Survival rates were recorded for 3 days after CLP, and all surviving rats were killed 3 days after CLP to examine their immune responses. RESULTS: Aerobic and anaerobic bacteria colony counts in peritoneal lavage fluid were significantly reduced, and the phagocytic activity of peritoneal macrophages was enhanced in groups 3 and 4 but not in the other 2 groups. There were no significant differences in the phagocytic activities of blood polymorphonuclear cells and survival rates among the 4 groups. CONCLUSIONS: Enteral Arg supplementation before sepsis significantly enhanced peritoneal macrophage phagocytic activity and reduced total bacterial counts in peritoneal lavage fluid. Arg administered before and after CLP seemed to have a synergistic effect on enhancing phagocytic activity and on bacterial clearance. However, IV Arg administration after CLP had no favorable effects on phagocytic activity or survival rates in rats with gut-derived sepsis. PMID- 12903886 TI - The effect of supplemental enteral glutamine on plasma levels, gut function, and outcome in severe burns: a randomized, double-blind, controlled clinical trial. AB - BACKGROUND: This research was conducted to evaluate the effect of enterally administered glutamine (gln) dipeptide on metabolic, gastrointestinal, and outcome parameters after severe burn injury. METHODS: Forty thermally injured patients with total body surface burns ranging between 50% and 80%, and third degree burns ranging between 20% and 40% and without respiratory injuries, were randomized into a prospective, double-blind, controlled clinical trial. One group received gln-enriched enteral nutrition and the other group received the standard enteral formulation. Tube feedings were initiated on postburn day 1 (PBD +1), and isocaloric and isonitrogenous feedings were administered to both groups until PBD +12. The gln was given as the dipeptide of alanyl-gln (Ajinomoto, Tokyo, Japan), which provided 0.35 g gln/kg body weight/d. Plasma amino acid profiles, serum endotoxin concentrations, and the lactulose/mannitol absorption ratio (which reflects gut permeability) were measured at specific times throughout the clinical course. Wound healing at day 30 was assessed, and length of hospital stay and total costs were determined at discharge. RESULTS: The 2 groups were similar in terms of age and extent of injury. Plasma gln concentrations were approximately 300 umol/L in both groups on PBD +1 and remained low in the control group (399 +/- 40 umol/L, mean +/- SD) but increased toward normal in the supplemented group to 591 +/- 74 (p = .048). Lactulose/mannitol ratios were increased above normal on POD +1 (control, 0.221 +/- 0.169; gln, 0.268 +/- 0.202; not significant), reflecting increased intestinal permeability after burn injury. On POD +3, the ratio in the gln group was lower than control (0.025 +/- 0.008 versus 0.049 +/- 0.016; p = .0001), and both groups returned toward normal ratios with time. Endotoxin levels on PBD +1 were elevated in both groups (control, 0.089 +/- 0.023 EU/mL; gln, 0.103 +/- 0.037 EU/mL; NS) but decreased significantly on PBD +3 in the patients receiving gln. Hospital stay was significantly shorter in the gln group than controls (67 +/- 4 days versus 73 +/- 6; p = .026). On day 30, wound healing was 86% +/- 2% complete in the gln group compared with 72% +/- 3% in controls (p = .041). Total cost of hospitalization was 62794 +/- 6178 RMB (dollar 7593 +/- 747 US dollars) in the gln group and 68996 +/- 8620RMB (dollar 8343 +/- 1042, p = .031) in controls, although the cost of the enteral nutrition was higher in the gln-supplemented patients. CONCLUSION: Enteral gln supplementation using a commercially available dipeptide supported plasma gln levels, improved gut permeability, and initially decreased plasma endotoxin levels in severely thermally injured patients. These alterations were associated with a reduction in the length of hospitalization and lower costs. PMID- 12903887 TI - Glutamine administration during total parenteral nutrition protects liver adenosine nucleotides during and after subsequent hemorrhagic shock. AB - BACKGROUND: Glutamine supplementation of total parenteral nutrition (TPN) in stressed patients has been advocated. To determine whether glutamine supplementation affects the host response to conditions of stress, animals were given TPN with or without glutamine for 7 days. They were then subjected to the acute stress of hemorrhagic shock, which results in marked loss of hepatic adenosine triphosphate (ATP) and adenosine diphosphate (ADP), with accumulation of adenosine monophosphate (AMP) and the metabolites adenosine, inosine, hypoxanthine, and xanthine. This loss of ATP and accumulation of metabolites contributes to subsequent tissue damage. The hypothesis of the study was that glutamine supplementation would significantly improve restoration of hepatic adenosine nucleotides before and after hemorrhagic shock. METHODS: Sprague-Dawley rats were given TPN for 7 days. One half of the animals (n = 8) received TPN supplemented with glutamine, while one half received TPN with an isonitrogenous mixture of alanine and glycine. Animals were subjected to hemorrhagic shock for 30 minutes and then resuscitated using only heparinized shed blood. Liver biopsies were taken pre- and post-shock, and at 30 and 60 minutes after resuscitation. ATP, ADP, AMP, and their metabolites were measured using gradient high-performance liquid chromatography. RESULTS: After 7 days of TPN, baseline values of ATP, ADP, AMP, and metabolites were similar between the 2 groups before the initiation of shock. Glutamine-treated animals manifested a 40% decrease in ATP level immediately after shock and recovered to 90% of baseline within 60 minutes. By contrast, the control animals manifested a 66% decrease in ATP level after the shock period and recovered only to 60% of baseline at 1 hour postresuscitation. Similar changes were observed in ADP levels and were accompanied by corresponding changes in AMP and adenosine metabolites, all of which rose during shock and fell after resuscitation. CONCLUSIONS: Glutamine supplementation significantly protected the liver from tissue damage caused by hemorrhagic shock. ATP levels remained higher during shock and recovered more rapidly after resuscitation. Glutamine supplementation may help to protect cellular energy stores in the stressed organism and may offer opportunities for therapeutic intervention during and after stress. PMID- 12903888 TI - Intestinal and multivisceral transplantation: dynamics of nutritional management and functional autonomy. AB - OBJECTIVE: The objective of this study was to describe the dynamics of nutrition management of intestinal transplant recipients and allograft functional autonomy. METHODS: Intestinal absorptive functions and recipient nutritional status were monitored during the 12-month study period. Absorption was evaluated with D xylose absorption and fecal fat excretion. Indices for nutrition were body weight, anthropometric measures, and serum albumin. RESULTS: Before transplant, all patients were total parenteral nutrition (TPN) dependent and well nourished. By the first postoperative month, all 22 recipients were tolerating enteral feeding. By 3 months, all recipients had begun oral feeding, with 13 off TPN and 7 off enteral feeds. By 6 months, 16 recipients were off TPN, and by the end of the 12th month, 17 (77%) were free of TPN. Although all 22 recipients were completely weaned off TPN during the first posttransplant year, 10 required temporary reinstitution of therapy at different points. Full nutritional autonomy was achieved at 3 months by 3 recipients, at 6 months by 8 recipients, and at 12 months by 12 (55%) recipients. CONCLUSIONS: These results reflect our early experience that led to surgical refinement of the operation and evolution of the recipient postoperative management. Nonetheless, even in this initial cohort, most of the engrafted intestines restored the recipient nutritional autonomy, and all survivors remained well nourished. PMID- 12903889 TI - Long-term effects of early postnatal food restriction on growth hormone secretion in rats. AB - BACKGROUND: Malnutrition in early life retards growth permanently in both humans and rats, but the underlying mechanism is unclear. The purpose of this study was to test the hypothesis that early postnatal food restriction induces long-term changes in the growth hormone-insulin-like growth factor I (GH-IGF-I) axis. METHODS: We examined the effect of increasing litter size to 20 during lactation [food restriction (FR)] on growth and spontaneous GH secretion and serum IGF-I levels in 14- to 15-week-old rats. RESULTS: No catch up in body weight (BW; p < .05), total length (TTL; p < .001), or tail length (TL; females, p < .02; males, p < .001) was observed in the adult female and male FR rats. Spontaneous 6-hour rat GH (rGH) secretory profiles showed significantly increased (p < .05) mean baseline rGH plasma concentrations in the male adult FR versus control (CON) rats (38.0 +/- 3.6 versus 26.4 +/- 2.5 ng/mL). Serum IGF-I levels in the male adult FR rat were significantly (p < .01) reduced (751.3 +/- 50.3 versus 985 +/- 55.5 ng/mL) compared with male controls. rGH secretory pattern in the adult female FR rat and serum IGF-I concentrations were not different from female controls. CONCLUSIONS: FR during lactation leads to incomplete catch-up growth in adult female and male rats. In the adult male FR rat, increased baseline rGH secretion and reduced serum IGF-I concentrations might explain the slow growth. PMID- 12903890 TI - Efficacy of voluntary nasogastric tube feeding in female inpatients with anorexia nervosa. AB - BACKGROUND: This investigation assesses the efficacy of a voluntary nasogastric tube feeding protocol on the weight gain of patients with anorexia nervosa, tube feeding's effect on recovery from the psychologic aspects of anorexia, patient satisfaction with treatment, and medical complications. METHODS: The study included a nonrandomized retrospective review of 381 female inpatients with a Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition diagnosis of anorexia nervosa, both subtypes. A total of 155 patients received tube feeding and oral refeeding; 226 received oral refeeding alone. Recovery from the psychologic aspects of anorexia was measured by the change in Eating Disorder Inventory-2 scores between admission and discharge. Patient satisfaction with treatment was measured with a patient satisfaction questionnaire completed at discharge. Repeated measures and multivariate analyses were performed. RESULTS: When severity-of-illness and caloric intake differences between patients with and without tube feeding were controlled, patients who received tube feeding gained significantly more weight per treatment week than those who received oral kilocalories alone. Patients who received tube feeding for at least one-half their length of stay gained 1 kg/week versus 0.77 kg/week for patients receiving oral refeeding alone. Tube-fed patients evidenced no differences in recovery from anorexia's psychologic aspects, satisfaction with treatment, or medical complication frequency. CONCLUSIONS: In residential psychiatric treatment settings in which intensive therapeutic interventions and appropriate medical monitoring can manage potential psychologic and medical risks, tube feeding's weight gain benefits may be a viable and safe option in treating anorexia. PMID- 12903891 TI - Gastric electrical stimulation for gastroparesis improves nutritional parameters at short, intermediate, and long-term follow-up. AB - BACKGROUND: Nutritional depletion, either macronutrient- or micronutrient related, is common in patients with drug-refractory gastroparesis (GP) for which there is often no effective treatment. We studied a group of 12 patients (4 men, 8 women; mean age, 35.7 years) who had symptoms of GP and were a subset of the Gastric ElectroMechanical Stimulation trial of gastric electrical stimulation. Patients' symptoms were of long duration (7.3 years) and associated with diabetes mellitus (3 patients) or were idiopathic (9 patients) as etiology. METHODS: The patients had permanent gastric electrical stimulation devices surgically implanted after a positive trial of temporary stimulation. Patients had baseline measures of gastrointestinal total symptom score (TSS), laboratory (albumin and related) measures, weight (kg), body mass index, and route of nutrition: oral feeding, enteral tubes, or hyperalimentation, repeated at 3, 6, and 12 months. Intermediate-term follow-up was done at 1 to 2 years, and long-term follow-up was done at 5 years and included gastrointestinal TSS, weekly vomiting frequency score, and nutrition and overall health-related quality-of-life measures. RESULTS: Gastric electrical stimulation was associated with rapid improvement over the short-term in TSS (35.6 at baseline to 16.6 at month 12; p < .05), body weight, body mass index, and serum albumin over the short term with most parameters improving by 3 to 6 months. Intermediate (1 to 2 years) and long-term (5 year) data showed continued improvement in TSS, vomiting frequency (weekly vomiting frequency score 3.9 at baseline to 1.7 at 5 years; p < .01), quality-of life measures, and maintenance of weight gain. CONCLUSIONS: Gastric electrical stimulation implantation resulted in improvement of nutritional parameters throughout the first 12 months, as nausea and vomiting decreased and oral intake increased. This improvement in nutritional measures is maintained long-term and is associated with improvements in quality of life. Gastric electrical stimulation should be considered as a therapeutic option for any patients with refractory GP and nutritional compromise. PMID- 12903892 TI - A survey of perceived benefit and differences in therapy provided by credentialed and noncredentialed nutrition support dietitians. AB - BACKGROUND: Benefits of the certified nutrition support dietitian (CNSD) credential to clinicians, their patients, and healthcare administrators have not been defined. A study was designed to measure the difference in cost of therapy provided by credentialed and noncredentialed nutrition support dietitians and to measure the perceived value of the credential to those who hold it. METHODS: Using a modified Delphi technique, a questionnaire was developed to obtain demographic information, responses to statements of perceived benefit of the credential, and costs of therapy selected for patients in common clinical scenarios. RESULTS: Of the 691 questionnaires mailed, 314 (45%) were returned. For 8 of 10 statements of perceived benefit, mean response scores indicated agreement. Respondents did not agree that salary increases resulted from credentialing. There was no difference between groups in the type of monitoring selected or in the time to complete an initial assessment. Cost of therapy was significantly higher for credentialed versus noncredentialed dietitians (dollar 915.67 +/- dollar 241.73 versus dollar 851.78 +/- dollar 243.44; p = .035), although the survey was not designed to show that the most appropriate care was least expensive. CONCLUSIONS: The CNSD credential is of perceived benefit to practitioners except in obtaining salary increases. Credentialed dietitians selected more expensive therapy than noncredentialed dietitians, which would benefit healthcare administrators only if reduction of other costs resulted. More complex models are needed to fully assess the benefit of credentialed nutrition support dietitians to patients and healthcare administrators. PMID- 12903893 TI - Cost and clinical effectiveness of certification cannot be measured using survey methods. PMID- 12903894 TI - Joint Commission for Accreditation of Health Care Organizations guidelines: too late to intervene for nutritionally at-risk surgical patients. AB - BACKGROUND: Joint Commission for Accreditation of Health Care Organizations (JCAHO) guidelines recommend nutritional screening within 24 hours of hospital admission, with full nutrition assessment if at-risk. This study examines whether these guidelines detect surgical patients who may benefit from preoperative nutrition support. METHODS: A retrospective chart review was conducted for patients undergoing nonemergent esophageal, gastric, pancreatic, or colon surgery between 1996 and 2001 to determine dates of admission, day of surgery, and timing of documented nutrition screening and assessment. RESULTS: Most patients undergo surgery within 24 hours of admission, and undergo initial screening or full assessment after surgery. Twenty percent of patients are admitted for diagnostic testing, preoperative cardiovascular workup, or medical management preoperatively and are assessed 5 to 6 days before surgery, but these are the minority. CONCLUSION: Because of the high incidence of nonemergent surgeries that occur within 24 hours of admission, screening and assessment by JCAHO guidelines cannot detect patients who may benefit from preoperative nutrition support. Systems to identify these patients must be implemented earlier in nutrition screening and assessment. PMID- 12903895 TI - Early developments and clinical applications of total parenteral nutrition. AB - BACKGROUND: This article recounts the conditions and status of surgical nutrition support in the 1960s and the antecedent basic and clinical investigational work leading to the development of a practical and efficacious method of adequate nourishment entirely by vein in Beagle puppies; describes the subsequent clinical application of the knowledge, techniques, and technology to the first successful long-term total parenteral nutrition (TPN) support of critically ill pediatric and adult patients; and admonishes nutritionists of all backgrounds that some need for parenteral nutrition will likely always exist and that it is incumbent upon everyone to continue endeavors to advance the germinal methodology to perfection. METHODS: The relevant indications, limitations, hindrances, motivational factors, and studies regarding the development of TPN are reviewed, and the fundamental investigational work culminating in the first successful growth and development of Beagle puppies and a human infant fed entirely by vein are described firsthand. The details of the orderly and logical scientific development of the principles and components of the techniques in animals, infants, and adults are related. RESULTS: Knowledge, techniques, and technologic constituents of the first successful long-term TPN system were developed in the basic biochemical and animal laboratories initially in 6 puppies and subsequently adapted clinically for the efficacious long-term i.v. support of 6 critically ill surgical adult patients and a newborn infant before its widespread clinical application. CONCLUSIONS: Long-term TPN was inaugurated successfully as a safe and effective i.v. feeding technique nearly 4 decades ago. However, basic and clinical investigations must continue to be encouraged, supported, and carried out in the quest to perfect the current rudimentary technology, methodology, and outcomes. PMID- 12903896 TI - The price of quality. PMID- 12903898 TI - A case of hypokalemia and rhabdomyolysis in a patient with short bowel syndrome. PMID- 12903897 TI - Glutamine to intensive care unit patients. PMID- 12903900 TI - Physiological adaptations to exercise in people with spinal cord injury. AB - The number of patients that suffer some type of spinal cord lesion in recent years are high and have increased because of factors such as traffic accidents. Although their life expectancy has increased, cardiovascular illnesses is one of the main causes of morbidity and mortality. Since the degree of physical fitness is an important factor regarding the risk of cardiovascular disease, the objective of the present study was to examine the global adaptation (cardiorespiratory, metabolic and thermoregulatory response) of the organism to exercise and the application of this data to the habitual practice of physical activity to improve state of health. A group of 42 patients with spinal injury, 85% of whom were paraplegic and the remaining 15% tetraplegic performed 42 exercise tests on a cycloergometer. Body temperature (tympanum, surface of the deltoids and surface of the back), metabolic parameters (plasma uric acid, glycemia, plasma lactate), cardiocirculatory adaptation (heart rate, blood pressure arm, blood pressure leg) and ventilatory adaptation (VO2, VCO2, fr Vt, VE) were monitored. Blood pressure in the arm, blood concentrations of lactate and ventilatory parameters showed an evolution statistically dependent on the work to which the subject was submitted. Heart rate showed a statistically significant correlation with the ventilatory parameters and work load. The proportional response of the cardioventilatory parameters to the increase in the work load allowed us to evaluate the repercussion of a given exercise and thus avoid exercise of an excessive intensity that could produce cardiocirculatory changes that might entail an added risk. Heart rate presents an excellent correlation, shown in this work, with the oxygen consumption and could therefore be used to quantify the cardiorespiratory and metabolic repercussion of the exercise carried out. Furthermore, this quantification may allow for the adaptation of exercise intensity to the patient thus improving the results obtained from the practice of exercise that has been proven so necessary in these patients. PMID- 12903899 TI - "2-6-11" motif in heat shock protein 60 and central nervous system antigens: a preliminary study in multiple sclerosis patients. AB - Multiple sclerosis (MS) is a chronic inflammatory demyelinating disease of the central nervous system (CNS) with unknown etiology and pathogenesis. A local autoimmune process involving activation of autoreactive T cells against CNS protein components is likely crucial in the development of MS lesions. Myelin reactive T cells are believed to be primed in the periphery during infections by antigens of bacterial or viral origin via molecular mimicry, a postulated mechanism that might account for the trigger of an autoimmune response on the basis of sequence homology between foreign and self determinants. Immune responses to heat shock proteins (hsp) have been implicated in the initiation or progression of a number of autoimmune diseases. Hsp may function as immunodominant targets during the immune response evoked by pathogens, and theoretically a cross-reactive response to sequences shared by these immunogens and autoantigens in the CNS may contribute to the pathogenesis of MS. We examined the immune response of peripheral blood mononuclear cells (PBMNc) from MS patients and healthy subjects elicited by peptides derived from hsp60 containing a common structural motif ("2-6-11" motif) already described, which is also present in CNS putative antigens. This structural pattern consists of an apolar residue or Lys at position 2, Pro always at position 6, and Glu, Asp or Lys at residue 11. Results reported here are indicative of maturation of peripheral blood monocytes towards a differentiated CD14(+)CD16(+)DR(+) cell and release of pro-inflammatory cytokines consistent with a Th1-like pattern. These are typical features exhibited by immune cells implicated in autoimmune responses. PMID- 12903901 TI - Effect of protein hydrolysis on the dialysability of amino acids and minerals in infant formulas. AB - The objectives of this study were to evaluate the dialysed quantities of amino acids and minerals (calcium, phosphorus, magnesium, iron and zinc) in two types of infant formulas, one with a basis of native cow milk proteins and the other highly hydrolysed, to evaluate the possible application of Miller's in vitro method in amino acid and mineral availability studies. The percentage of dialysis differed between proteins. The hydrolysis treatment applied to the proteins significantly increased the dialysis percentage of almost all the amino acids. The dialysability of all the minerals was statistically greater in the formula made with a basis of hydrolysed proteins. Miller's method was seen to be of use for showing the effect which the hydrolysis treatment has on the availability of amino acids. Despite its limitations of having to be carried out in vitro, the study suggests that the nutritional value in relation with amino acids, minerals and trace elements differs between the both types of formulas. PMID- 12903902 TI - [Ca2+]i changes in guinea pig tracheal smooth muscle cells in culture: effects of Na+ and ouabain. AB - The objective of this work was to confirm that the contractile effects of ouabain and Na(+)-free solutions in guinea pig tracheal rings are associated with increments in the cytosolic free Ca2+ concentration ([Ca2+]i) in cultured tracheal smooth muscle (TSM) cells. Cultured cells were alpha-actin positive. Histamine (50 microM) and Na(+)-free solution elicited a transient increase in [Ca2+]i, while the responses to thapsigargin (1 microM) and ouabain (1 mM) were long lasting. However, carbachol (10, 200, and 500 mM) and high K(+)-solution produced no effect on [Ca2+]i, suggesting that cultured guinea pig TSM cells display a phenotype change but maintain some of the tracheal rings physiological properties. The transient rise in [Ca2+]i in response to the absence of extracellular Na+ and the effect of ouabain may indicate the participation of the Na(+)/Ca2+ exchanger (NCX) in the regulation of [Ca2+]i. PMID- 12903903 TI - Identification of 100 KDa protein in sera of mice-treated with Cu(II) complex with superoxide dismutase-mimetic activity. AB - The protein profile of sera isolated from mice pre-treated with Cu(II) complex of Girard T with superoxide dismutase (SOD)-mimetic activity was analyzed using SDS PAGE. This complex was intraperitoneally administered (10 mg/Kg body weight) to Swiss albino mice. The resolved polypeptides showed a new sharp band at 100 KDa against which a polyclonal antibody was raised in rabbit. Sera of rabbit anti-100 KDa protein was used as a powerful probe for the detection of 100 KDa protein isolated from sera of treated mice. Western blot assays revealed a strong reactive polypeptide band at 100 KDa in sera of the mice, but no cross reaction was observed with sera of normal mice. The identification of purified polypeptide was confirmed by different characterization experiments. PMID- 12903904 TI - Effect of cadmium on lymphocyte subsets distribution in thymus and spleen. AB - This work was designed to analyze the possible dose dependent effects of cadmium on the distribution of lymphocyte subsets within the thymus and spleen. Cadmium accumulation was also evaluated in these tissues. For this purpose, adult male rats were exposed for one month to 0, 5, 10, 25, 50 or 100 ppm of cadmium chloride (CdCl2) in the drinking water. In both spleen and thymus, the B lymphocytes increased with the doses of 5 and 10 ppm of CdCl2, and decreased with the doses of 25-100 ppm. In spleen, the doses of 25 and 50 ppm decreased CD4+ cells and the doses of 5 and 10 ppm increased CD8+ cells, while the percentage of thymus T, CD4+, CD8+ and CD4(+)-CD8+ cells was not modified by cadmium treatment at any dose used in this study. After cadmium exposure, the metal was accumulated in the spleen only from the dose of 50 ppm on, and in the thymus, from the dose of 10 ppm on. In conclusion, although the accumulation of the metal is higher in thymus than in spleen, the metal affected CD4+ and CD8+ lymphocytes at the spleen but not at the thymus. PMID- 12903906 TI - Effects of S-adenosylmethionine on hepatic regeneration after partial hepatectomy in the rat. PMID- 12903907 TI - Semmelweis' discovery and its Finnish follow-up. AB - Professor Ignac Semmelweis (1818-1865) is one of the great personalities of medical history. He insisted on washing hands with chlorine water before any obstetrical intervention, he was the first to demonstrate its importance in preventing puerperal fever. Thus, the principle of asepsis was introduced prior to the discovery of bacteria and bacterial diseases. Semmelweis carefully documented his findings and in this way pioneered the scientific analysis of clinical data Medical community of that time misinterpreted Semmelweis' great ideas, he died abandoned and forgotten. A Finnish doctor Josef Adam Joachim Pippingskold was one of the first obstetricians who had realized the importance of Semmelweis' work. In 1861, in his letter to Semmelweis he reported about his own findings and favorable results in prevention of puerperal fever in Helsinki. Two decades earlier, Dr. Ehrstrom in the University of Helsinki had submitted his thesis on pathophysiology of puerperal fever that was similar to the ideas of Semmelweis. Long before modern times in Finland, mothers traditionally had their babies delivered in smoke saunas, where heating and smoke of bactericidal phenols created a clean, rather aseptic environment. Hand washing was self-evident necessity. However, the situation was quite different in the Central European universities and departments of obstetrics, where the medical training and clinical practice took place side by side. Semmelweis' life and his contribution to medicine was appreciated even in the theatrical circles of Finland. The piece "Semmelweis" of Norwegian playwright Jens Bjorneboe got its World Premier in the Swedish Theatre in Turku, former capital of Finland, in September 1969. PMID- 12903905 TI - Adipose tissue as an endocrine organ: role of leptin and adiponectin in the pathogenesis of cardiovascular diseases. AB - Obesity, the most common nutritional disorder in industrial countries, is associated with increased cardiovascular mortality and morbidity. Nevertheless, the molecular basis linking obesity with cardiovascular disturbances have not yet been fully clarified. Recent advances in the biology of adipose tissue indicate that it is not simply an energy storage organ, but also a secretory organ, producing a variety of bioactive substances, including leptin and adiponectin, that may influence the function as well as the structural integrity of the cardiovascular system. Leptin, besides being a satiety signal for the central nervous system and to be related to insulin and glucose metabolism, may also play an important role in regulating vascular tone because of the widespread distribution of functional receptors in the vascular cells. On the other hand, the more recently discovered protein, adiponectin, seems to play a protective role in experimental models of vascular injury, in probable relation to its ability to suppress the attachment of monocytes to endothelial cells, which is an early event in the atherosclerotic process. There is already considerable evidence linking altered production of some adipocyte hormones with the cardiovascular complications of obesity. Therefore, the knowledge of alterations in the endocrine function of adipose tissue may help to further understand the high cardiovascular risk associated with obesity. PMID- 12903908 TI - Effect of orally and intraperitoneally administered plant lectins on food consumption of rats. AB - A panel of orally administered lectins (100 mg/kg b.w.) of different binding specificities was tested for suppression of voluntary food consumption in prefasted rats. PHA isolectins (Phaseolus vulgaris) and RPA-I (Robinia pseudoacacia) were found to exert a marked and significant effect, but two other gut-binding lectins, i.e. SBA (Glycine max) and WGA (Triticum vulgar) and several non-binding lectins were ineffective. In cannulated rats PHA infused into the duodenum induced food suppression, i.e. binding of the lectin to the mouth or stomach was unnecessary. Suppression of food consumption lasted through the whole nocturnal feeding period, control (BSA) and experimental (PHA) curves of cumulative food consumption showed a V-like divergence. Suppression by PHA or RPA I showed very similar time courses, but a long-lasting inhibition of gastric emptying was only observed in the RPA-treated animals. Intraperitoneally administered lectins suppressed food consumption much more effectively than the oral ones, whereas Galanthus nivalis agglutinin (ONA) had little or no effect. It is concluded that lectins can be used as effective tools for the modulation of food consumption and gastric emptying in experimental animals. PMID- 12903909 TI - Oesophageal acid stimulation in humans: does it alter baroreflex function? AB - The aim of this study was to investigate if oesophagel acid stimulation (Bernstein test) had an influence on heart rate and blood prsure variability and baroreflex gain. We compared the cardiovascular responses in 10 patients with established gastro-esophageal reflux disease (Group 1) and 10 control subjects (Group 2) during esophageal saline and 0.1 mol/l hydrochloric acid instillation. Indices of heart rate and blood pressure variability and baroreflex gain (derived from linear spontaneous sequences and cross spectral analysis) were calculated. In Group 1 the standard deviation of RR intervals (SDRR: 46 ms vs 51 ms, p=0.030) and the root mean square of successive differences (RMSSD: 24 ms vs. 26 ms p=0.027) were significantly lower during acid infusions, than during saline. We found no significant difference in minimum, maximum and mean RR intervals and systolic blood pressures and in the percentage of RR intervals, which differed from adjacent cycles by more than 50 ms (PNN50). The power spectra of RR intervals in the high frequency band tended to be lower during acid infusion (p=0.055). There was no significant difference in blood pressure spectra, neither in low nor in high frequency band. In Group 2 there was no significant difference between any parameters measured during acid and saline. The baroreflex gain was not changed during the studied conditions in any group. Neither increased vagal tone, nor increased vagal variability occurred and the baroreflex gain was not altered during oesophageal acid simulation. PMID- 12903910 TI - Activity of cathepsin B, D and L in rat cerebrum after cimetidine and famotidine administration. AB - Cathepsins are lysosomal enzymes that are used a sensitive markers in various toxicological investigations. The purpose of this study was to evaluate and compare the influence of cimetidine and famotidine on the cerebral cortex, particularly on the activity of cortical cathepsin B, D and L in the frontal lobe of rat brain. The drugs were administered intraperitoneally, twice a day, for six weeks to male Wistar rats in two doses. The initial dose was 2.85 mg/kg for cimetidine and 0.285 mg/kg for famotidine. The second dose was 10 times higher. Control animals were injected with 0.9% NaCl. Half of the animals from each of the drug-treated and control groups were sacrificed on the 42nd day of the experiment. The remaining animals were raised for another 6 weeks without any xenobiotics, and sacrificed on the 84th day. The frontal lobe of the right cerebral hemisphere was taken for biochemical investigation. The activities of free and bound fractions of cathepsin B, D and L were evaluated spectrophotometrically in cortical homogenates. The activity of bound fraction of cathepsin D and L decreased significantly in animals exposed to the higher dose of cimetidine and sacrificed on the 42nd day. Also significant elevation of the free fraction of cathepsin L was noted in the same group of rats. Cathepsin activities were normalized during the next six weeks. No behavioural changes were noted among the observed animals. Unlike cimetidine, famotidine did not change profiles of the cerebral cathepsins. PMID- 12903911 TI - Effects of zinc deficiency and supplementation on some hematologic parameters of rats performing acute swimming exercise. AB - The aim of the study was to investigate how zinc deficiency and supplementation effect some hematologic parameters of rats performing swimming exercise. Forty adult male Spraque-Dawley rats were divided into 4 groups, zinc deficient swimming group (Group 1, n=10, zinc supplemented swimming group (Group 2, n=10), swimming control group (Group 3, n=10), and control group (Group 4, n=10). Blood samples were taken by decapitation and analyzed for the determination of erythrocyte, hemoglobin level, hematocrit, leukocyte, lymphocyte, platelet count and plasma zinc level at the end of the 4 week experiment. Erythrocyte count of group 1 was the lowest whereas erythrocyte count in group 3 was significantly lower than that in group 2 and 4 (p<0.05). Hemoglobin level of group 1 was significantly lower than that of groups 2 and 4 (p<0.05). Hematocrit was significantly lower in both group 1 and group 3 than both groups 2 and 4 (p<0.05). Lymphocyte count in group 2 was significantly higher than in all other groups (p<0.05). Platelet counts in group 2 was significantly lower than in all other groups (p<0.05). Our findings suggest that zinc deficiency effects the hematologic parameters mentioned negatively whereas zinc supplementation has a positive influence. PMID- 12903913 TI - Interaction of noscapine with the bradykinin mediation of the cough response. AB - Angiotensin Converting Enzyme Inhibitors (ACEI) like captopril and enalapril, can induce persistant cough in man. Noscapine, an antitussive alkaloid, can be used to suppress ACEI-induced cough. Some workers have suggested a role for bradykinin in precipitation of ACE-induced cough. Work carried out in our laboratory has shown noscapine to be a non-competitive inhibitor of bradykinin in guinea pig ileum. It is therefore possible that noscapine suppresses cough by blocking the effect of bradykinin receptor activation in the airways. Guinea pigs were placed in a cough-chamber connected to an air pump and a pressure transducer. Capsaicin was sprayed into the chamber and cough was recorded as a distinctive change in air pressure inside the cough-chamber. Animals treated with 1 mg/kg captopril and enalapril for 7 days, showed increased cough response. Ten microgram/kg FR190997, a non-peptide agonist of the bradykinin B2 receptor, also increased the cough response. Noscapine at 0.5, 1 and 2 mg/kg was able to reverse the effects of ACEI and FR190997. Naloxone, a specific opioid receptor inhibitor, did not block the antitussive effects of noscapine in enalapril or FR190997 treated guinea pigs. This antitussive effect of noscapine is not mediated via the mu, kappa or delta opioid receptors. It is therefore possible that noscapine exerts its antitussive action by interfering with the bradykinin cough mediation. PMID- 12903912 TI - The effect of adrenomedullin, amylin fragment 8-37 and calcitonin gene-related peptide on contractile force, heart rate and coronary perfusion pressure in isolated rat hearts. AB - The effect of human adrenomedullin, human amylin fragment 8-37 (amylin 8-37) and rat calcitonin gene-related peptide (CGRP) on contractile force, heart rate and coronary perfusion pressure has been investigated in the isolated perfused rat hearts. Adrenomedullin (2x10(-10), 2x10(-9) and 2x10(-8) M) produced a significant decrease in contractile force and perfusion pressure, but only the peptide caused a decline in heart rate at the highest dose. Amylin (10(-9), 10( 8) and 10(-7) M) significantly increased and then decreased contractile force. Two doses of amylin (10(-8) and 10(-7) M) induced a significant increase in heart rate, however amylin did not change perfusion pressure in all the doses used. Rat alpha CGRP (10(-8), 10(-7) and 10(-6) M) evoked a slight decline in contractile force following a significant increase in contractile force induced by the peptide. CGRP in all the doses raised heart rate and lowered perfusion pressure. Our results suggest that adrenomedullin has negative inotropic, negative chronotropic and coronary vasodilator actions. Amylin produces a biphasic inotropic effect and evokes a positive chronotropy. CGRP causes positive inotropic, positive chronotropic and vasodilatory effects in isolated rat hearts. PMID- 12903914 TI - Dead-ended autologous connective tissue chambers in peripheral nerve repair- early observations. AB - The effects of the repair of nerve gap injuries are still unsatisfactory, despite the great progress in microsurgery. Until now, there is no effective method to induce the regeneration of the transected peripheral nerve when its distal stump is missing. The aim of this work was to examine whether the implantation of dead ended connective tissue chambers can promote the outgrowth of injured peripheral neurites. This method differs from all previous nerve guides because it totally eliminates the distal part of the nerve and restricts the influence of surrounding tissues. We have also tried to establish whether some neurotrophic factors can be applied by means of these chambers. The results of this work show that dead-ended autologous connective tissue chambers can be a useful tool in peripheral nerve injuries treatment, even when the distal part of the nerve is missing. PMID- 12903915 TI - The teaching of physiological disciplines in the 3rd Faculty of Medicine, Charles University, Prague, Czech Republic. AB - The physiological disciplines are being taught from the 1st year of medical study up to the end of the 4th year, it means 8 semesters altogether. It is necessary to explain the curriculum of the 3rd Faculty of Medicine, Charles University in Prague. This Faculty has had a new curriculum since 1997, so the first students, who had been taught as to this system, already finished their study. The duration of medical study is 6 years, i.e. 12 semesters. This composed of three cycles: I Basic biomedical sciences. The first two years represent the first cycle, which is based on the integrative principle. II. Principles of clinical medicine. The second cycle is concerned on the problem-based learning. III. Clinical preparation. The third cycle represents clinical application. In the first two years the integrative study is composed of different modules. The first module I A--Structure and functions of human body is the greatest module in the first two years, which is composed of anatomy, histology and embryology, biochemistry and physiology. The module I B--Cell biology and genetics is composed of genetics and cell biology and this module is finished by the examination after two semesters at the end of the first year. PMID- 12903916 TI - Coagulation monitoring and management during liver transplantation. PMID- 12903917 TI - Newly developed T-wave inversion with cardiac wall-motion abnormality predominantly occurs in middle-aged or elderly women after noncardiac surgery. AB - PURPOSE: The study was done to determine the characteristics and prevalence of myocardial ischemia with inverted T waves after noncardiac surgery. METHODS: A list of patients who developed electrocardiogram (ECG) T-wave inversion associated with wall-motion abnormalities observed by transthoracic echocardiography (TTE) following noncardiac surgery was generated from the intensive care unit (ICU) medical records database between January 1, 1995, and December 31, 2000. The hospital records of these patients were analyzed retrospectively. RESULTS: Among 4219 patients (2187 men and 2032 women) who were admitted to the ICU after noncardiac surgery, 13 developed myocardial ischemia with inverted T waves postoperatively. All of the patients were middle-aged or elderly women with no history of coronary artery disease; nine of them had undergone intraabdominal surgery. Characteristic ECG findings included inverted T waves in the left precordial leads, which subsequently became prominent with QT prolongation. In all of these patients, wall-motion abnormalities were observed on the anterior wall, but these resolved within 60 days of the episode. Myocardial ischemia was asymptomatic, with neither hemodynamic changes nor adverse cardiac events. CONCLUSION: Newly developed giant negative T waves with QT prolongation in the ECG may indicate myocardial stunning, but do not in themselves imply a poor prognosis. The marked preponderance of middle-aged and elderly women with this type of myocardial ischemia remains to be explained. PMID- 12903918 TI - Reduction of potential respiratory pathogens by oral hygienic treatment in patients undergoing endotracheal anesthesia. AB - PURPOSE: This study was conducted to evaluate the usefulness of mechanical and chemical prophylactic oral cleansing treatments for reducing potential respiratory pathogens existing in the oral cavity. METHODS: Thirty-two patients scheduled to undergo oral and maxillofacial surgery that required endotracheal anesthesia were randomly allocated to one of the two groups, the oral cleansing group (n = 16) or the noncleansing group (n = 16). Culture and polymerase chain reaction (PCR) methods were used to detect and enumerate pathogens. Oral cleansing was carried out with an electric toothbrush capable of automatically supplying and aspirating povidone-iodine solution before surgery, followed by rinsing twice a day after surgery. Cephazolin (3 g x day(-1)) was given to all patients for 5 days after surgery. RESULTS: The PCR detection rates of Streptococcus pneumoniae, Haemophilus influenzae, Pseudomonas aeruginosa, and Porphyromonas gingivalis in gargle samples before treatment were 87.5%, 68.8%, 53.1%, and 40.6%, respectively. Oral cleansing reduced the detection rates and numbers of methicillin-sensitive Staphylococcus species, S. pneumoniae, and H. influenzae. In contrast, there was no significant reduction of methicillin resistant Staphylococcus species, S. pneumoniae, H. influenzae, or P. aeruginosa in subjects who underwent systemic cephazolin administration without oral cleansing. CONCLUSION: The combination of mechanical and chemical oral cleansing resulted in a significant reduction of potential respiratory pathogens in the oral cavity. PMID- 12903919 TI - MAC reduction after intrathecal coadministration of GABA(A) agonist and glutamate antagonist in rats. AB - PURPOSE: It has been reported that intrathecal coadministration of a GABA(A) agonist and a glutamate antagonist induces synergistic antinociceptive effects in rats. We hypothesized that this synergistic antinociceptive effect might induce a synergistic reduction in the minimum alveolar anesthetic concentration (MAC). METHODS: The MAC for sevoflurane was determined before and after intrathecal administration of muscimol (GABA(A) agonist, 0.1-10 microg), AP-5 (N-methyl-D aspartate [NMDA] antagonist, 0.1-10 microg), and YM872 (alpha-amino-3-hydroxy-5 methyl-4-isoxazole propionic acid [AMPA] antagonist, 0.1-10 microg) in rats. The effects of coadministration of muscimol and AP-5 or YM872 on MAC reduction were also tested. RESULTS: Intrathecal administration of muscimol at doses of 1 and 10 microg significantly reduced the MAC by 34.5% +/- 3.3% and 46.9% +/- 4.4%, respectively (P < 0.01). Intrathecal administration of AP-5 at a dose of 10 microg and YM872 at a dose of 10 microg significantly reduced the MAC by 25.8% +/ 2.4% and 31.4% +/- 6.3%, respectively (P < 0.01). No additional reductions in MAC values were observed when a GABA(A) agonist was combined with either an NMDA or an AMPA antagonist. CONCLUSION: Intrathecal coadministration of a GABA(A) agonist and an NMDA or AMPA antagonist, which has been reported to produce a synergistic antinociceptive effect, did not induce a synergistic reduction in the MAC. The antinociceptive effect may not be the predominant factor in the reduction of the MAC induced by these drugs. PMID- 12903920 TI - Comparative fading responses induced by mivacurium, cisatracurium, and d tubocurarine in the evoked muscular compound action potentials of the cat. AB - PURPOSE: It has been suggested that the different degrees of fade induced by nondepolarizing neuromuscular blocking agents in repetitive muscular contractions may be due to the varying affinities or binding kinetics of presynaptic nicotinic receptors. We compared the degree of fade induced by mivacurium, cisatracurium, and d-tubocurarine in the cat muscular compound action potential (mCAP). METHODS: In 21 cats, mCAPs of the gastrocnemius muscle were evoked by paired (conditioning and test stimuli) and 2 Hz train-of-four (TOF) sciatic nerve stimulation. The interval between the paired stimuli was changed stepwise from 7 to 1000 ms. The ratios of the amplitude evoked by test stimulus to that evoked by the conditioning stimulus (M2/M1 ratios) and TOF ratios were measured. After baseline variables had been obtained, the cat received either mivacurium (0.08 mg x kg( 1), n = 7), cisatracurium (0.05 mg x kg(-1), n = 7), or d-tubocurarine (0.5 mg x kg(-1), n = 7). A series of M2/M1 ratios and TOF ratios were measured at various levels of partial block during recovery. RESULTS: At 10% of baseline amplitude, all agents significantly depressed the M2/M1 ratios (i.e., fade) at relatively longer intervals of paired stimuli (mivacurium, > or = 100 ms; cisatracurium. > or = 40 ms; and d-tubocurarine, > or = 20 ms), when compared with baseline. The order of activity to produce fade was mivacurium < cisatracurium < d tubocurarine. A similar result was obtained in TOF ratios measured at various levels of neuromuscular block. CONCLUSION: Our results suggest that mivacurium shows a lesser degree of fade during partial neuromuscular block than cisatracurium and d-tubocurarine. PMID- 12903922 TI - Deep temperature monitoring using a zero-heat-flow method. PMID- 12903921 TI - Effects of ketamine on neostigmine-induced contractile and phosphatidylinositol responses of the rat trachea. AB - PURPOSE: Neostigmine causes airway smooth muscle contraction through the direct stimulation of muscarinic receptors and the activation of phosphatidylinositol (PI) responses. Ketamine attenuates airway smooth muscle contraction. It is not clear whether ketamine attenuates neostigmine-induced airway smooth muscle contraction by inhibiting the PI response. This study was designed to examine the effects of ketamine on neostigmine-induced contractile and PI responses of the rat trachea. METHODS: Thirty male Wistar rats weighing 250-350 g were used. In the experiment on the contractile response, active contraction was induced with 1 microM neostigmine in the presence or absence of ketamine. In the experiment on the phosphatidylinositol response, the trachea slices were incubated with [3H]myo inositol, 1 microM neostigmine, or 100 microM aluminum fluoride, and ketamine. The formation of [3H]inositol monophosphate (IP1), a degradation product of the phosphatidylinositol response, was measured with a liquid scintillation counter. Statistical significance (P < 0.05) was determined by analysis of variance. RESULTS: Neostigmine 1 microM caused tracheal ring contraction. This contraction was attenuated by ketamine dose-dependently and reached resting tension at 100 microM. Neostigmine- and aluminum fluoride-induced IP1, accumulation was also attenuated by ketamine. CONCLUSION: The results suggest that ketamine attenuates neostigmine-induced contractile responses, at least in part, through the inhibition of phospholipase C coupled with G protein in the PI response. PMID- 12903923 TI - Anesthetic principles in living-donor liver transplantation at Kyoto University Hospital: experiences of 760 cases. PMID- 12903924 TI - Itch is an unpleasant sensory experience. PMID- 12903925 TI - Comparison of the effects of neuroleptanesthesia and enflurane or sevoflurane anesthesia on neuromuscular blockade by rocuronium. PMID- 12903926 TI - Effects of cuff pressure on changes in airway morphology after use of the laryngeal mask airway. PMID- 12903927 TI - Hemodynamic effects of nasal continuous positive airway pressure after abdominal surgery. PMID- 12903929 TI - Evaluation of thalamic neural activity in CRPS type 1 patients by proton MR spectroscopy: a correlative study with rCBF. PMID- 12903928 TI - Effect of isolation stress on anesthetic requirement in mice. PMID- 12903930 TI - Effect of humidity in the circuit on the CO2 absorption capacity of Amsorb and Sodasorb II. PMID- 12903931 TI - Propofol pharmacokinetics in a patient with bilateral leg amputation. PMID- 12903932 TI - Central neuraxial blockade in ambulatory surgery. PMID- 12903933 TI - Sedation with propofol for 33 days in a patient with acute aortic valve regurgitation and dissecting aneurysm. PMID- 12903934 TI - Application of NMR spectroscopy and LC-NMR/MS to the identification of carbohydrates in beer. AB - The application of LC-NMR/MS for the direct identification of carbohydrates in beer has been studied. Carbohydrates are major beer components, and their structural characterization by NMR alone is seriously hindered by strong spectroscopic overlap. Direct analysis of beer by LC-NMR/MS enables the rapid (1 2 h) identification of dextrins with degree of polymerization (DP) of up to nine monomers, with degassing being the only sample treatment required. Although the presence of alpha(1-->6) branching points is easily indicated by NMR for each subfraction separated by LC, difficulties arise for the unambiguous assignment of linear or branched forms of high DP dextrins. The two beer samples investigated in this work were found to have significantly different oligosaccharide compositions, reflecting the different production conditions employed. The use of hyphenated NMR for the rapid characterization of the carbohydrate composition of beers may be the basis of a useful tool for the quality control of beer. PMID- 12903935 TI - Extraction of chili, black pepper, and ginger with near-critical CO2, propane, and dimethyl ether: analysis of the extracts by quantitative nuclear magnetic resonance. AB - Ginger, black pepper, and chili powder were extracted using near-critical carbon dioxide, propane, and dimethyl ether on a laboratory scale to determine the overall yield and extraction efficiency for selected pungent components. The temperature dependency of extraction yield and efficiency was also determined for black pepper and chili using propane and dimethyl ether. The pungency of the extracts was determined by using an NMR technique developed for this work. The volatiles contents of ginger and black pepper extracts were also determined. Extraction of all spice types was carried out with acetone to compare overall yields. Subcritical dimethyl ether was as effective at extracting the pungent principles from the spices as supercritical carbon dioxide, although a substantial amount of water was also extracted. Subcritical propane was the least effective solvent. All solvents quantitatively extracted the gingerols from ginger. The yields of capsaicins obtained by supercritical CO(2) and dimethyl ether were similar and approximately double that extracted by propane. The yield of piperines obtained by propane extraction of black pepper was low at approximately 10% of that achieved with dimethyl ether and CO(2), but improved with increasing extraction temperature. PMID- 12903936 TI - Simultaneous liquid chromatographic determination of creatinine and pseudouridine in bovine urine and the effect of sample pH on the analysis. AB - A rapid, reliable method for the simultaneous determination of creatinine and pseudouridine is described. Both analytes were detected at an optimum wavelength of detection (262 nm), considering the relative levels present in bovine urine. Cimetidine was used as the internal standard and detected at its maximum wavelength of absorption (220 nm) on a separate channel. All three compounds were eluted within 15 min, using a 10 mmol/L phosphate buffer (pH 6.8)-methanol gradient on a C18 column. Creatinine data were found to be significantly dependent upon the pH of the sample. Recoveries of both analytes were above 96%. Lowest detectable levels of creatinine and pseudouridine were 0.28 nmol and 9.0 pmol, respectively. The use of internal standard resulted in a method with high precision (standard deviation of 1.42 mmol/L and 0.027 mmol/L for creatinine and pseudouridine), yet one that was simple and rapid. PMID- 12903937 TI - Differentiation of natural and synthetic phenylalanine and tyrosine through natural abundance 2H nuclear magnetic resonance. AB - The natural abundance deuterium NMR characterization of samples of the amino acids tyrosine (1) and phenylalanine (2), examined as the acetylated methyl esters 4 and 6, has been performed with the aim to identify by these means the contribution in animals of the hydroxylation of the diet l-phenylalanine (2) to the formation of l-tyrosine (1), a feature previously revealed on the same samples through the determination of the phenolic delta(18)O values. The study, which includes also the NMR examination of benzoic acid (5) from 2 and of tyrosol (7) from 1, substantially fails in providing the required information because the mode of deuterium labeling of tyrosine samples of different origins is quite similar but indicates a dramatic difference in the deuterium labeling pattern of the two amino acids 1 and 2. The most relevant variation is with regard to the deuterium enrichments at the CH(2) and CH positions, which are inverted in the two amino acids of natural derivation. Moreover, whereas the diastereotopic benzylic hydrogen atoms of l-tyrosine (1) appear to be equally deuterium enriched, in l-phenylalanine (2) the (D/H)(3)(R)() > (D/H)(3)(S)(). Similarly, benzoic acid (5) shows separate signals for the aromatic deuterium nuclei, which are quite indicative of the natural or synthetic derivation. The mode of deuterium labeling of the side chain of 1 and 2 is tentatively correlated to the different origins of the two amino acids, natural from animal sources for l tyrosine and biotechnological probably from genetically modified microorganisms for l-phenylalanine. PMID- 12903938 TI - Identification of volatile compounds in soybean at various developmental stages using solid phase microextraction. AB - Soybean (Glycine max) seed volatiles were analyzed using a solid phase microextraction (SPME) method combined with gas chromatography-mass spectrometry (GC-MS). Thirty volatile compounds already reported for soybean were recovered, and an additional 19 compounds not previously reported were identified or tentatively identified. The SPME method was utilized to compare the volatile profile of soybean seed at three distinct stages of development. Most of the newly reported compounds in soybean seed were aldehydes and ketones. During early periods of development at maturity stage R6, several volatiles were present at relatively high concentrations, including 3-hexanone, (E)-2-hexenal, 1-hexanol, and 3-octanone. At maturity stage R7 and R8, decreased amounts of 3-hexanone, (E) 2-hexenal, 1-hexanol, and 3-octanone were observed. At maturity stage R8 hexanal, (E)-2-heptenal, (E)-2-octenal, ethanol, 1-hexanol, and 1-octen-3-ol were detected at relatively high concentrations. SPME offers the ability to differentiate between the three soybean developmental stages that yield both fundamental and practical information. PMID- 12903939 TI - Isotopic labeling and LC-APCI-MS quantification for investigating absorption of carotenoids and phylloquinone from kale (Brassica oleracea). AB - The ability to study bioavailability of nutrients from foods is an important step in determining the health impact of those nutrients. This work describes a method for studying the bioavailability of nutrients from kale (Brassica oleracea var. Acephala) by labeling the nutrients with carbon-13, feeding the kale to an adult volunteer, and analyzing plasma samples for labeled nutrients. Results showed that conditions for producing atmospheric intrinsically labeled kale had no detrimental effect on plant growth. Lutein, beta-carotene, retinol, and phylloquinone were analyzed using liquid chromatography-atmospheric pressure chemical ionization mass spectrometry. Analysis of plasma samples showed that labeled lutein peaked in plasma at 11 h (0.23 microM), beta-carotene peaked at 8 (0.058 microM) and 24 h (0.062 microM), retinol peaked at 24 h (0.10 microM), and phylloquinone peaked at 7 h (3.0 nM). This method of labeling kale with (13)C was successful for producing clearly defined kinetic curves for (13)C-lutein,(13)C beta-carotene, (13)C-retinol, and (13)C-phylloquinone. PMID- 12903940 TI - Ovicidal and adulticidal effects of Eugenia caryophyllata bud and leaf oil compounds on Pediculus capitis. AB - The toxicity of Eugenia caryophyllata bud and leaf oil-derived compounds (acetyleugenol, beta-caryophyllene, eugenol, alpha-humulene, and methyl salicylate) and congeners of eugenol (isoeugenol and methyleugenol) against eggs and females of Pediculus capitis was examined using direct contact application and fumigation methods and compared with those of the widely used delta phenothrin and pyrethrum. In a filter paper diffusion bioassay with female P. capitis, the pediculicidal activity of the Eugenia bud and leaf oils was comparable to those of delta-phenothrin and pyrethrum on the basis of LT(50) values at 0.25 mg/cm(2). At 0.25 mg/cm(2), the compound most toxic to female P. capitis was eugenol followed by methyl salicylate. Acetyleugenol, beta caryophyllene, alpha-humulene, isoeugenol, and methyleugenol were not effective. Eugenol at 0.25 mg/cm(2) was as potent as delta-phenothrin and pyrethrum but was slightly less effective than the pyrethroids at 0.125 mg/cm(2). Against P. capitis eggs, methyl salicylate and eugenol were highly effective at 0.25 and 1.0 mg/cm(2), respectively, whereas little or no activity at 5 mg/cm(2) was observed with the other test compounds as well as with delta-phenothrin and pyrethrum. In fumigation tests with female P. capitis at 0.25 mg/cm(2), eugenol and methyl salicylate were more effective in closed cups than in open ones, indicating that the effect of the compounds was largely due to action in the vapor phase. Neither delta-phenothrin nor pyrethrum exhibited fumigant toxicity. The Eugenia bud and leaf essential oils, particularly eugenol and methyl salicylate, merit further study as potential P. capitis control agents or lead compounds. PMID- 12903941 TI - Anthocyanin and proanthocyanidin content in selected white and red wines. Oxygen radical absorbance capacity comparison with nontraditional wines obtained from highbush blueberry. AB - Antioxidant capacity, as measured by oxygen radical absorbance capacity (ORAC(PE)), total phenolic, total and individual anthocyanins, and proanthocyanidin fraction contents were evaluated in red and white wines from grapes. A comparison in terms of antioxidant capacity is made with nontraditional wines made from highbush blueberry. Blueberries are among fruits that are best recognized for their potential health benefits. In red wines, total oligomeric proanthocyanidin content, including catechins, was substantially higher (177.18 +/- 96.06 mg/L) than that in white wines (8.75 +/- 4.53 mg/L). A relative high correlation in red wines was found between ORAC(PE) values and malvidin compounds (r = 0.75, P < 0.10), and proanthocyanidins (r = 0.87, P < 0.05). In white wines, a significant correlation was found between the trimeric proanthocyanidin fraction and peroxyl radical scavenging values (r = 0.86, P < 0.10). A moderate drink (1 drink per day, about 140 mL) of red wine, or white wine, or wine made from highbush blueberry corresponds to an intake of 2.04 +/- 0.81 mmol of TE, 0.47 +/- 0.15 mmol of TE, and 2.42 +/- 0.88 mmol of TE of ORAC(PE)/day, respectively. PMID- 12903942 TI - Isolation and antihypertensive effect of angiotensin I-converting enzyme (ACE) inhibitory peptides from spinach Rubisco. AB - Four new inhibitory peptides for angiotensin I-converting enzyme (ACE), that is, MRWRD, MRW, LRIPVA, and IAYKPAG, were isolated from the pepsin-pancreatin digest of spinach Rubisco with the use of HPLC. IC(50) values of individual peptides were 2.1, 0.6, 0.38, and 4.2 microM, respectively. MRW and MRWRD had an antihypertensive effect after oral administration to spontaneously hypertensive rats. Maximal reduction occurred 2 h after oral administration of MRW, whereas MRWRD showed maximal decrease 4 h after oral administration at doses of 20 and 30 mg/kg, respectively. IAYKPAG also exerted antihypertensive activity after oral administration at the dose of 100 mg/kg, giving a maximum decrease 4 h after oral administration. IAYKP, IAY, and KP, the fragment peptides of IAYKPAG, also exerted antihypertensive activity. LRIPVA [corrected] did not show any antihypertensive effect at a dose of 100 mg/kg despite its potent ACE-inhibitory activity. PMID- 12903943 TI - Chemical characterization and biological effects of Sicilian Opuntia ficus indica (L.) mill. Fruit juice: antioxidant and antiulcerogenic activity. AB - The juice of whole fruits of Sicilian cultivars of prickly pear (Opuntia ficus indica (L.) Mill.) was investigated, and the contents of ascorbic acid, total polyphenols, and flavonoids were determined. In the juice, ferulic acid was the chief derivative of hydroxycinnamic acid and the mean concentration of total phenolic compounds was 746 microg/mL. The flavonoid fraction, analyzed by high performance liquid chromatography-diode array detection, consisted of rutin and isorhamnetin derivatives. The juice showed antioxidant activity in the DPPH(*) test, probably due to the phenolic compounds that are effective radical scavengers. The preventive administration of the juice inhibited the ulcerogenic activity of ethanol in rat. Light microscopy observations showed an increase in mucus production and the restoration of the normal mucosal architecture. The juice is nutritionally interesting, and its dietary intake could provide protection against oxidative damage. PMID- 12903944 TI - Enhancing volatile phenol concentrations in wine by expressing various phenolic acid decarboxylase genes in Saccharomyces cerevisiae. AB - Phenolic acids, which are generally esterified with tartaric acid, are natural constituents of grape must and wine and can be released as free acids (principally p-coumaric, caffeic, and ferulic acids) by certain cinnamoyl esterase activities during the wine-making process. Some of the microorganisms present in grape can metabolize the free phenolic acids into 4-vinyl and 4-ethyl derivatives. These volatile phenols contribute to the aroma of wine. The Saccharomyces cerevisiae phenyl acrylic acid decarboxylase gene (PAD1) is steadily transcribed, but its encoded product, Pad1p, shows low activity. In contrast, the phenolic acid decarboxylase (PADC) from Bacillus subtilis and the p coumaric acid decarboxylase (PDC) from Lactobacillus plantarum display substrate inducible decarboxylating activity in the presence of phenolic acids. In an attempt to develop wine yeasts with optimized decarboxylation activity on phenolic acids, the padc, pdc, and PAD1 genes were cloned under the control of S. cerevisiae's constitutive phosphoglyceratekinase I gene promoter (PGK1(P)()) and terminator (PGK1(T)()) sequences. These gene constructs were integrated into the URA3 locus of a laboratory strain of S. cerevisiae, Sigma1278b. The overexpression of the two bacterial genes, padc and pdc, in S. cerevisiae showed high enzyme activity. However, this was not the case for PAD1. The padc and pdc genes were also integrated into an industrial wine yeast strain, S. cerevisiae VIN13. As an additional control, both alleles of PAD1 were disrupted in the VIN13 strain. In microvinification trials, all of the laboratory and industrial yeast transformants carrying the padc and pdc gene constructs showed an increase in volatile phenol formation as compared to the untransformed host strains (Sigma1278b and VIN13). This study offers prospects for the development of wine yeast starter strains with optimized decarboxylation activity on phenolic acids and the improvement of wine aroma in the future. PMID- 12903945 TI - Molluscicidal saponins from Sapindus mukorossi, inhibitory agents of golden apple snails, Pomacea canaliculata. AB - Extracts of soapnut, Sapindus mukorossi Gaertn. (Sapindaceae) showed molluscicidal effects against the golden apple snail, Pomacea canaliculata Lamarck. (Ampullariidae) with LC(50) values of 85, 22, and 17 ppm after treating 24, 48, and 72 h, respectively. Bioassay-directed fractionation of S. mukorossi resulted in the isolation of one new hederagenin-based acetylated saponin, hederagenin 3-O-(2,4-O-di-acetyl-alpha-l-arabinopyranoside)-(1-->3)-alpha-l rhamnopyranosyl-(1-->2)-alpha-l-arabinopyranoside (1), along with six known hederagenin saponins, hederagenin 3-O-(3,4-O-di-acetyl-alpha-l-arabinopyranoside) (1-->3)-alpha-l-rhamnopyranosyl-(1-->2)-alpha-l-arabinopyranoside (2), hederagenin 3-O-(3-O-acetyl-beta-d-xylopyranosyl)-(1-->3)-alpha-l-rhamnopyranosyl (1-->2)-alpha-l-arabinopyranoside (3), hederagenin 3-O-(4-O-acetyl-beta-d xylopyranosyl)-(1-->3)-alpha-l-rhamnopyranosyl-(1-->2)-alpha-l-arabinopyranoside (4), hederagenin 3-O-(3,4-O-di-acetyl-beta-d-xylopyranosyl)-(1-->3)-alpha-l rhamnopyranosyl-(1-->2)-alpha-l-arabinopyranoside (5), hederagenin 3-O-beta-d xylopyranosyl-(1-->3)-alpha-l-rhamnopyranosyl-(1-->2)-alpha-l-arabinopyranoside (6), and hederagenin 3-O-alpha-l-arabinopyranoside (7). The bioassay data revealed that 1-7 were molluscicidal, causing 70-100% mortality at 10 ppm against the golden apple snail. PMID- 12903946 TI - Cynodontin: a fungal metabolite with antifungal properties. AB - A red pigment that accumulates in cultures of a Drechslera avenae pathotype with specificity for Avena sterilis was isolated and identified as the anthraquinone cynodontin (3-methyl-1,4,5,8-tetrahydroxyanthraquinone). Satisfactory yield of the compound was obtained with 20-60 day incubations at temperatures between 20 and 27 degrees C. Cynodontin was tested in vitro for fungitoxicity and was found to be a potent inhibitor of the growth of Sclerotinia minor, Sclerotinia sclerotiorum, and Botrytis cinerea and, to a lesser extent, of Verticillium dahliae. The ED(50) values obtained with these fungi were of the same order of magnitude as those of the commercial fungicides dicloran and carbendazim, which were used as reference chemicals. In contrast, the growth of a number of other fungi was not significantly inhibited by cynodontin. Anthraquinone and two other anthraquinone derivatives, emodin and chrysophanol, which were also included in the tests, did not affect the growth of the cynodontin-sensitive fungi. It thus appears that the type and position of the substitutions at the C-ring play a role in the expression of antifungal activity. PMID- 12903947 TI - Isolation and characterization of some antioxidative compounds from the rhizomes of smaller galanga (Alpinia officinarum Hance). AB - Antioxidative compounds were isolated from the methanol extract of fresh rhizome of smaller galanga (Alpinia officinarum Hance). Seven phenylpropanoids (1-7) were finally obtained by reversed-phase HPLC, and their structures were elucidated by MS and NMR analyses. They comprised the two known compounds, (E)-p-coumaryl alcohol gamma-O-methyl ether (1) and (E)-p-coumaryl alcohol (6), and the five novel compounds, stereoisomers of (4E)-1,5-bis(4-hydroxyphenyl)-1-methoxy-2 (methoxymethyl)-4-pentene (2a and 2b), stereoisomers of (4E)-1,5-bis(4 hydroxyphenyl)-1-ethoxy-2-(methoxymethyl)-4-pentene (3a and 3b), (4E)-1,5-bis(4 hydroxyphenyl)-1-[(2E)-3-(4-acetoxyphenyl)-2-propenoxy]-2-(methoxymethyl)-4 pentene (4), (4E)-1,5-bis(4-hydroxyphenyl)-2-(methoxymethyl)-4-penten-1-ol (5), and (4E)-1,5-bis(4-hydroxyphenyl)-2-(hydroxymethyl)-4-penten-1-ol (7). Compounds 1-7 were detected for the first time as constituents of galanga rhizomes and exhibited antioxidative activities against the autoxidation of methyl linoleate in bulk phase. PMID- 12903948 TI - Angiotensin I converting enzyme (ACE) inhibitory activity of hetero chitooligosaccharides prepared from partially different deacetylated chitosans. AB - Angiotensin I converting enzyme (ACE) inhibitory activity of hetero chitooligosaccharides (hetero-COSs) prepared from partially different deacetylated chitosans was investigated. Partially deacetylated chitosans, 90, 75, and 50% deacetylated chitosan, were prepared from crab chitin by N deacetylation with 40% sodium hydroxide solution for durations. In addition, nine kinds of hetero-COSs with relatively high molecular masses (5000-10 000 Da; 90 HMWCOSs, 75-HMWCOSs, and 50-HMWCOSs), medium molecular masses (1000-5000 Da; 90 MMWCOSs, 75-MMWCOSs, and 50-MMWCOSs), and low molecular masses (below 1000 Da; 90 LMWCOSs, 75-LMWCOSs, and 50-LMWCOSs) were prepared using an ultrafiltration membrane bioreactor system. ACE inhibitory activity of hetero-COSs was dependent on the degree of deacetylation of chitosans. 50-MMWCOSs that are COSs hydrolyzed from 50% deacetylated chitosan, the relatively lowest degree of deacetylation, exhibited the highest ACE inhibitory activity, and the IC(50) value was 1.22 +/- 0.13 mg/mL. In addition, the ACE inhibition pattern of the 50-MMWCOSs was investigated by Lineweaver-Burk plots, and the inhibition pattern was found to be competitive. PMID- 12903949 TI - Discrimination of three trachurus species using both mitochondrial- and nuclear based DNA approaches. AB - A double-DNA approach was developed to discriminate the three Trachurus species that abide in European waters: T. trachurus, T. mediterraneus, and T. picturatus. The analysis aimed at both mitochondrial and nuclear loci. Polymerase Chain Reaction (PCR) amplification of the cytochrome b gene of mtDNA was followed by restriction analysis with three species-specific enzymes: NlaIII, NciI, and BsmAI. Digestion with these endonucleases yielded species-specific electrophoretic profiles. The universality of the results was verified by screening a large number of individuals from 12 geographical regions covering most of the distribution of the species. Additionally, the nuclear multicopy 5S rRNA gene was selected as an alternative candidate for the discrimination of the three Trachurusspecies. A simple agarose gel electrophoretic analysis of the amplicons proved to be capable of leading to unambiguous identification of the three Trachurus species. Thus, the double-DNA methodology presented here allows the accurate discrimination of Trachurus fish species and the detection of commercial fraud. PMID- 12903950 TI - Investigation of packaging systems for shelled walnuts based on oxygen absorbers. AB - Storage of nuts at a high oxygen concentration results in rancid nuts whereas storage at a low oxygen concentration results in fine-tasting nuts. During a 13 month experiment, packaging of walnuts with an oxygen absorber was compared to packaging in nitrogen or atmospheric air. At the same time, the effects of oxygen permeability of the packaging material and storage temperature (11 and 21 degrees C) were investigated by determination of hexanal and rancid taste of the walnuts. The optimal storage condition for walnuts is at 11 degrees C or lower, eventually combined with an oxygen absorber. However, without chilled storage and use of an oxygen absorber, it is possible to obtain an acceptable quality of walnuts with a packaging material having a very low oxygen permeability (e.g., laminate with EVOH) combined with nitrogen flushing. The results also revealed that the development of hexanal during time can be described by a second-order polynomial regression model. PMID- 12903951 TI - Concentrations of low and high molecular weight thiols in wheat dough as affected by different concentrations of ascorbic acid. AB - Different amounts of ascorbic acid (AA) were added to flour, and the concentrations of low and high molecular weight thiols in the dough were determined. For the determination of the low molecular weight thiols, glutathione, cysteine, and the corresponding disulfides, an isotope dilution assay with a (14)C-labeled internal standard was used. For the determination of the high molecular weight thiols, a method was developed that involved derivatization of dough with Ellman's reagent, removal of excess reagent by dialysis, micro-Osborne fractionation, release of the label by reduction, and determination of reduced Ellman's reagent by reversed-phase high-performance liquid chromatography. Mixing of flour without AA led to a decrease of the glutathione and an increase of the cysteine concentration. Addition of AA reduced the concentration of both thiols to a minimum when 125 mg of AA/kg of flour was applied. Furthermore, the concentrations of high molecular weight thiols in the glutenins of flours from different wheat cultivars were determined. The values ranged from 5.6 to 8.2 micromol/kg of protein and showed a correlation between flour quality and SH concentration. On addition of AA and mixing of a dough, the concentrations of the protein thiols in the glutenins isolated from the dough increased to a maximum when 100 mg of AA/kg of flour was added. Higher concentrations of AA led to a decrease of the SH concentration. The last results are not in accordance with previously published data or with current hypotheses about the mechanism of the AA improver action. PMID- 12903952 TI - Effect of ascorbic acid in dough: reaction of oxidized glutathione with reactive thiol groups of wheat glutelin. AB - The reactions of oxidized glutathione generated from endogenous glutathione by the addition of ascorbic acid (AA) prior to dough mixing on free thiol groups of gluten proteins have been investigated. A small amount of (35)S-labeled glutathione was added as a tracer to identify the reaction products of GSSG and free protein thiols by radioactivity measurement. First, gluten was isolated from the dough, then the gliadins were extracted, and residual glutenin was partially hydrolyzed with thermolysin. After preseparation by gel permeation chromatography, the fractions with the highest radioactivity were separated by high-performance liquid chromatography. Radioactive peptides were identified, isolated, sequenced, and assigned to amino acid sequences of gluten protein components. The isolated peptides contained exclusively the cysteine residues C(b) and C(x) of low molecular weight subunits of glutenin, which are supposed to be highly reactive in forming intermolecular disulfide bonds. From these results it can be assumed that the cysteine residues C(b) and C(x) of the low molecular weight subunits of glutenin are at least partly present in the thiol form in flour. During dough mixing they are converted to protein-protein disulfides or glutathione-protein mixed disulfides by thiol/disulfide interchange reactions. Oxidized glutathione necessary for this reaction is generated from glutathione by the action of AA. These results are in accordance with the major hypothesis about the mechanism of action of AA. PMID- 12903953 TI - Status of free radicals in Indian monsooned coffee beans gamma-irradiated for disinfestation. AB - Free radicals in two cultivars of Indian monsooned coffee beans, gamma-irradiated for hygienic and quarantine purposes, were examined by entrapping the small amount of samples in potassium chloride powder in ESR quartz tubes. In contrast to a prominent free radical signal at g = 2.002, observed in spermoderm (silver skin) and cotyledon (whole seed without skin) parts of normal coffee beans, the same was not discernible in monsooned coffee bean parts of both cultivars. The ESR signal was found to be more prominent in the spermoderm than in the whole seed portion of the normal coffee beans. Common practices of roasting and powdering were found to generate quantitatively more free radicals in coffee beans than gamma-irradiation alone. Phenols, contributing maximally to observed free radical signals in coffee beans, were significantly different in normal and monsooned coffee beans. These observations on insignificant free radical population in irradiated monsooned coffee beans may be attributed to their inherent possession of high water activity, favoring decay of free radicals produced. Textural studies with monsooned coffee beans, before and after mild heat treatments, supported these findings. PMID- 12903954 TI - Lipids from flax fibers and their fate in alkaline pulping. AB - The chemical composition of lipids from bast fibers of flax (Linum usitatissimum), which are commonly used for high-quality paper pulp production, was thoroughly studied by gas chromatography-mass spectrometry. The main compounds identified were waxes, series of long chain n-fatty alcohols, n aldehydes, n-fatty acids, and n-alkanes. Free and esterified sterols and triterpenols, steroid hydrocarbons, steroid and triterpenoid ketones, as well as sterol glycosides were also found in the flax bast fibers. On the other hand, the fate of these lipophilic compounds in alkaline pulping of flax fibers was investigated by analyzing two pulps obtained under distinct industrial cooking conditions. The results revealed that while waxes could be efficiently hydrolyzed during pulping depending on the alkali charge, most of the other lipophilic compounds present in flax fibers survived cooking and were present in the unbleached pulps. PMID- 12903955 TI - Influences of gamma-irradiation and storage on the carotenoids of sun-dried and dehydrated paprika. AB - The effects of drying methods, gamma-irradiation, and storage on the carotenoids (capsanthin, capsorubin, zeaxanthin, capsolutein, violaxanthin, beta-carotene, and beta-cryptoxanthin) of paprika were investigated. Sun-dried and dehydrated paprika samples were irradiated in a (60)Co gamma-irradiator at five doses (0, 2.5, 5.0, 7.5, and 10 kGy) in polyethylene bags and stored for 10 months at ambient temperature. Individual carotenoid analyses were carried out on the paprika and fresh red pepper during a 2 month period using the reverse phase HPLC technique. The concentrations of capsanthin and capsorubin, which are responsible for the attractive red color of sun-dried paprika, were higher than those of dehydrated paprika. Higher irradiation doses and a longer storage period resulted in a significant (P < 0.01) reduction of all the carotenoids, except capsorubin. There was no significant (P < 0.05) effect of irradiation dose on capsorubin destruction. The decrease of red carotenoids, for all irradiation treatments, was less than that of the storage period. Even the highest irradiation dose, 10 kGy, caused a 11.1% capsanthin reduction; however, 10 months of storage at the ambient temperature caused a 42.1% reduction of capsanthin. Yellow pigments of paprika (zeaxanthin, capsolutein, violaxanthin, beta-carotene, and beta-cryptoxanthin) were significantly (P < 0.01) decreased by all treatments. These yellow pigments were also found to be at high levels in those sun-dried samples with red pigments. This difference could be caused by the contribution of pigment biosynthesis during the sun-drying period. The most significant pigment reduction was realized in the processing and storage conditions of paprika, rather than in the irradiation process. PMID- 12903956 TI - Analysis of some Italian lemon liquors (limoncello). AB - The chemical composition of several commercial Italian Limoncellos, lemon-peel based alcoholic beverages, was studied by chromatographic techniques. These methods allowed a rapid monitoring of Limoncello, giving information on quality markers and possible adulteration of the product. Quantitative data for more than 60 compounds are reported. Limoncellos were characterized by the presence of selected volatile (terpenes, aldehydes, alcohols) and nonvolatile compounds (psoralens, coumarins, phenolics, carbohydrates and acids). On the basis of their composition, the samples were grouped by PCA analysis in two sets; the first group showed a composition similar to lemon essential oils, with a high content of b-pinene, myrcene, trans-a-bergamottene, and b-bisabolene, and a low content in neral and geranial. The composition of the second group suggested the occurrence of oxidative phenomena and/or the addition of flavors. The presence of ethyl acetate, acetaldehyde, 2-methyl-1-propanol and glycerol showed that a fermentation probably occurred in the sugar syrup used to dilute the Limoncello after the extraction process. PMID- 12903957 TI - Apoplastic pH and monolignol addition rate effects on lignin formation and cell wall degradability in maize. AB - Monolignol polymerization rate and apoplastic pH and may influence the formation of lignin and its interactions in cell walls. Primary maize walls were artificially lignified by gradual "end-wise" or rapid "bulk" polymerization of coniferyl alcohol at pH 4 or 5.5. Lignification efficiency was greatest for end wise polymers at pH 5.5 (90-98%), intermediate for bulk polymers formed at either pH (54-82%), and lowest for end-wise polymers at pH 4 (41-53%). End-wise polymers had about 2.2-fold more ether inter-unit linkages and 70% fewer end-groups than bulk polymers. Low pH enhanced the formation of ether linkages in end-wise but not in bulk polymers. Differences in lignin structure did not influence the enzymatic degradability of cell walls, but lowering apoplastic pH from 5.5 to 4.0 during lignification reduced cell wall degradability by 25%. Further studies indicated this pH-dependent depression in degradability was related to cell wall cross-links formed via lignin quinone methide intermediates. PMID- 12903958 TI - Origin of the color of Cv. rhapsody in blue rose and some other so-called "blue" roses. AB - Flowers of the rose cultivar Rhapsody in Blue display unusual colors, changing as they age, from a vivid red-purple to a lighter and duller purple, which are based on tonalities corresponding to hue angles between 340 and 320 degrees in the CIELAB scale. Unexpectedly, the chemical basis of these colors is among the simplest, featuring cyanin (cyanidin 3,5-di-O-glucoside), the most frequent anthocyanin in flowers, as the sole pigment and quercetin kaempferol glycosides as copigments at a relatively low copigment/pigment ratio (about 3/1), which usually produces magenta or red shades in roses. This color shift to bluer shades is coupled with the progressive accumulation of cyanin into vacuolar anthocyanic inclusions (AVIs), the occurrence of which increases as the petals grow older. In addition to the normal lambda(max) of cyanin at approximately 545 nm, the transmission spectra of live petals and of epidermal cells exhibit a second lambda(max) in the 620-625 nm range, the relative importance increasing with the presence of AVIs. In petals of fully opened flowers, the only pigmented structures in the vacuoles of epidermal cells are AVIs; their intense and massive absorption in the 520-640 nm area produces a much darker and bluer color than measured for the vacuolar solution present at the very first opening stage. Cyanin is probably "trapped" into AVIs at higher concentrations than would be possible in a vacuolar solution and in quinonoidal form, appearing purple-blue because of additional absorption in the 580-630 nm area. Quite similar pigmentation features were found in very ancient rose cultivars (cv. L'Eveque or Bleu Magenta), also displaying this type of so-called "blue" color. PMID- 12903959 TI - Alkaloidal components in the poisonous plant, Ipomoea carnea (Convolvulaceae). AB - Natural intoxication of livestock by the ingestion of Ipomoea carnea (Convolvulaceae) sometimes occurs in tropical regions of the world. Polyhydroxylated alkaloids were isolated from the leaves, flowers, and seeds of the poisonous plant and characterized. Chromatographic separation of the leaf extract resulted in the isolation of swainsonine (1), 2-epi-lentiginosine (2), calystegines B(1) (3), B(2) (4), B(3) (5), and C(1) (6), and N-methyl-trans-4 hydroxy-l-proline (7). The contents of 1 in the fresh leaves and flowers were 0.0029 and 0.0028%, respectively, whereas the contents of 1, 3, and 4 in the seeds were approximately 10 times higher than those in the leaves and flowers. Alkaloids 3, 4, and 6 showed a potent inhibitory activity toward rat lysosomal beta-glucosidase, with IC(50) values of 2.1, 0.75, and 0.84 microM, respectively, and alkaloid 5 was a moderate inhibitor of alpha- and beta-mannosidases. Although alkaloid 1 is known as a powerful inhibitor of lysosomal alpha-mannosidase (IC(50) = 0.02 microM), alkaloid 2, which has been thought to be an intermediate in the biosynthesis of 1, was also a potent inhibitor of alpha-mannosidase with an IC(50) value of 4.6 microM. PMID- 12903960 TI - The most abundant protease inhibitor in potato tuber (cv. Elkana) is a serine protease inhibitor from the Kunitz family. AB - The gene of the most abundant protease inhibitor in potato cv. Elkana was isolated and sequenced. The deduced amino acid sequence of this gene showed 98% identity with potato serine protease inhibitor (PSPI), a member of the Kunitz family. Therefore, the most abundant protease inhibitor was considered to be one of the isoforms of PSPI. The PSPI group represents approximately 22% of the total amount of proteins in potato cv. Elkana and is composed of seven different isoforms that slightly differ in isoelectric point. Antibodies were raised against the two most abundant isoforms of PSPI. The binding of these antibodies to PSPI isoforms and protease inhibitors from different groups of protease inhibitor in potato showed that approximately 70% of the protease inhibitors present in potato juice belong to the Kunitz family. PMID- 12903962 TI - Interaction between fenhexamid and yeasts during the alcoholic fermentation of Saccharomyces cerevisiae. AB - The behavior of the fungicide fenhexamid, N-(2,3-dichloro-4-hydroxyphenyl)-1 methyl-cyclohexanecarboxamide, has been studied at concentrations corresponding to the limits fixed for grapes (3 mg kg(-1)), or higher, during the alcoholic fermentation. The presence of the fungicide did not affect the amount of alcohol produced. The amount of fenhexamid in the liquid phase decreased by ca. 15%, but the missing fenhexamid was recovered unchanged from yeasts. This suggests that the fungicide is not degraded during the fermentation process, but adsorbed by yeasts. Two constituents of Saccharomyces cerevisiae cell wall, chitin and glucan, tested as potential adsorbents, exhibited affinity for fenhexamid. PMID- 12903961 TI - Identification of flavonol and xanthone glycosides from mango (Mangifera indica L. Cv. "Tommy Atkins") peels by high-performance liquid chromatography electrospray ionization mass spectrometry. AB - Flavonol O- and xanthone C-glycosides were extracted from mango (Mangifera indica L. cv. "Tommy Atkins") peels and characterized by high-performance liquid chromatography-electrospray ionization mass spectrometry. Among the fourteen compounds analyzed, seven quercetin O-glycosides, one kaempferol O-glycoside, and four xanthone C-glycosides were found. On the basis of their fragmentation pattern, the latter were identified as mangiferin and isomangiferin and their respective galloyl derivatives. A flavonol hexoside with m/z 477 was tentatively identified as a rhamnetin glycoside, which to the best of our knowledge, has not yet been reported in mango peels. The results obtained in the present study confirm that peels originating from mango fruit processing are a promising source of phenolic compounds that might be recovered and used as natural antioxidants or functional food ingredients. PMID- 12903963 TI - Metabolism of fenitrothion and conjugation of 3-methyl-4-nitrophenol in tomato plant (Lycopersicon esculentum). AB - The metabolism of (14)C-labeled fenitrothion (Sumithion, [O,O-dimethyl-O-(3 methyl-4-nitrophenyl)phosphorothioate]) in tomato plant (Lycopersicon esculentum Mill., cv. Ponderosa) grown in the greenhouse equipped with quartz glass was conducted to investigate the effect of sunlight on the behavior of fenitrothion and to elucidate the detailed structure of conjugated metabolites. Tomato plants (BBCH 85) were topically treated with (14)C-labeled fenitrothion twice with a 2 week interval between applications. At 15 days after the second application, more than half of the recovered (14)C was detected as unaltered fenitrothion, glucose, and cellobiose esters of 3-methyl-4-nitrophenol (NMC) in extracts from tomato fruit. The photoinduced formation of the S-methyl isomer of fenitrothion via thiono-thiolo rearrangement was detected only in the surface rinse but at trace amounts. In the whole tomato fruit, fenitrothion, the S isomer, NMC-beta glucoside, and NMC cellobioside were detected at 34.16, 1.28, 7.47, and 15.07% of the recovered (14)C, respectively. Trace amounts of the oxon analogue of fenitrothion were detected only on tomato leaves. The chemical structure of the cellobiose conjugate of NMC, 1-O-beta-d-glucopyranosyl-(1-->4)-beta-d glucopyranosyl-3-methyl-4-nitrophenol, was determined by spectroscopic analyses (liquid chromatography-mass spectrometry, NMR), using the metabolite obtained from leaves and stems of tomato plant hydroponically grown with (14)C-labeled NMC. PMID- 12903964 TI - Organochlorine pesticide residues in leek (Allium porrum) crops grown on untreated soils from an agricultural environment. AB - Leek (Allium porrum) plants from organic farming were harvested at 15, 59, and 210 days after seed germination. Organochlorine pesticide (OCP) levels were quantified by GC-ECD in vegetative tissues (roots and aerial), bulk soil and rhizosphere. Leek plant bioaccumulate OCPs efficiently in their aerial and root tissues and alter the concentration of the soil where they are grown. OCPs distribution pattern of bulk soil was endosulfans > DDTs > dieldrin, while it was endosulfans > HCHs > DDTs in leek tissues. There were statistically significant declines in DDTs, chlordane, dieldrin, and heptachlor in the rhizosphere, indicating that recalcitrant residues of OCPs may be removed from contaminated soil using leek crops under normal growing conditions. The DDE/DDT and alpha /gamma-HCH ratios of < 1 would indicate recent inputs of DDT and lindane in the environment. The occurrence of OCPs in this farm could be the result of atmospheric deposition and/or surface runoff of these pesticides. PMID- 12903965 TI - Synthesis and herbicidal activity of 2-cyano-3-substituted pyridinemethylaminoacrylates. AB - Two series of 2-cyano-3-substituted-pyridinemethylaminoacrylates, namely 12 new (Z)-2-cyano-3-methylthio-3-substituted-pyridinemethaneaminoacrylates and 10 new (Z)-2-cyano-3-alkyl-3-substituted-pyridinemethaneaminoacrylates, were synthesized as herbicidal inhibitors of photosystem II (PSII) electron transport. All of these compounds were confirmed by (1)H NMR, elemental, IR, and mass spectrum analyses. Their herbicidal activities were evaluated. Some compounds exhibited excellent herbicidal activities, even at a dose of 75 g/ha. A suitable substituent at the 2-position of the pyridine ring and the well-fit group at the 3-position of acrylate were essential for high herbicidal activity. 2 Cyanoacrylates containing a substituted pyridine ring provide higher herbicidal activities than parent compounds containing phenyl. These PSII inhibitor herbicides are safe to corn, which is a major crop in China. PMID- 12903966 TI - Complexation of several fungicides with beta-cyclodextrin: determination of the association constants and isolation of the solid complexes. AB - The formation of inclusion complexes with beta-cyclodextrin was studied for several popular fungicides of different types: prochloraz, 2-phenylphenol, thiophanate methyl, 8-hydroxyquinoline, and benalaxyl. Phase solubility diagrams showed that in all cases complexation takes place, leading to an important increase of water solubility in prochloraz and benalaxyl. Equilibrium association constants could be determined from the phase solubility data and from NMR titrations in the case of 2-phenylphenol. Because of the low solubility of the complex formed between 8-hydroxyquinoline and beta-cyclodextrin, the corresponding association constant could not be determined. The solid complexes of fungicide-cyclodextrin were prepared and isolated by different methods. The isolation of real complexes and not physical mixtures was confirmed in the cases of prochloraz, 2-phenylphenol, and benalaxyl by differential scanning calorimetry. PMID- 12903967 TI - Procedure for the measurement of soil inputs of plant-protection agents washed off through vineyard canopy by rainfall. AB - Soil inputs produced by rainfall waters washed off through a Ribeiro vineyard (Galicia, northwestern Spain) confirmed that levels recovered are <5% of the added fungicides. Laboratory studies showed that some fungicides were degraded in the ODS cartridge used for the collection of fungicides washed off by rainfall. Procymidone wash-off data obtained in the vineyard can be explained by a first order rate mass transfer model. A procedure for the collection of wash-off waters based on octadecylsilane solid phase extraction, followed by an analytical method of reverse-phase liquid chromatography with diode array detection is presented for the evaluation of fungicide inputs into soils after being washed off vineyards by rainfalls. Quality parameters of the analytical method yielded good precision (RSD < 10%) and low detection limits (ranging between 1 and 21 microg/L). PMID- 12903968 TI - Pesticide adsorptivity of aged particulate matter arising from crop residue burns. AB - Particulates (ashes) arising from the burning of crop residues are potentially effective adsorbents for pesticides in agricultural soils. To determine the long term adsorptive sustainability of ashes, a wheat (Triticum aestivum L.) ash was aged under environmentally relevant conditions (in CaCl(2) solution at room temperature and pH 7) in soil extract for 1 month and in a soil (1% ash) for a period of up to 12 months. The aged ash and ash-amended soil were used to sorb diuron from water. The diuron sorption was also measured in the presence of atrazine as a competing pesticide. There was no observed microbial impact on the stability of the wheat ash in soil. All isotherms with the ash were nonlinear type-I curves, suggestive of the surface adsorption. On a unit mass basis, the ash in soil extract was 600-10000 times more effective than the soil in sorbing diuron. Adsorption of dissolved soil organic matter (DOM) during aging on the ash surfaces reduced the diuron adsorption by 50-60%. Surface competition from the atrazine adsorption also reduced the ash adsorption of diuron by 10-30%. A total of 55-67% reduction in diuron sorption by the ash-amended soil was observed. Due to its high initial adsorptivity, the ash fraction of the aged ash-amended soil contributed >50% to the total diuron sorption. Thus, the wheat ash aged in the soil remained highly effective in adsorbing diuron. As crop residues are frequently burned in the field, pesticides in agricultural soils may be highly immobilized due to the presence of ashes. PMID- 12903969 TI - Simultaneous real-time measurements of mastication, swallowing, nasal airflow, and aroma release. AB - Mastication, swallowing, breath flow, and aroma release were measured simultaneously in vivo using electromyography, electroglottography, a turbine air flow meter, and the MS-Nose, respectively. Signals were synchronized either electronically or by aligning the nasal airflow data with the breath by breath release of acetone. Chewing affected nasal airflow, with the flow fluctuations following the mastication pattern. Data analysis suggested that air was pumped out of the mouth into the throat with each chew, and the mean volume was 26 mL. Aroma release was associated with the pulses of air pumped from the mouth with each chew. During swallowing, there was no nasal airflow, but after swallowing, aroma release was evident. The volume of the retronasal route was estimated at 48 mL when swallowing and 72 mL when samples were chewed. The combination of techniques shows the effects of physiological processes on aroma release. PMID- 12903970 TI - Dynamic flavor release from sucrose solutions. AB - The initial dynamic flavor release from sucrose solutions was modeled. Modeling was based on the theoretical hydration behavior of sucrose, theoretical physicochemical data of flavor volatiles, and process parameters of a headspace apparatus used for model validation. The rate-limiting factor determining the initial flavor release was the hydration of sucrose, which in turn depends on the molarity of sucrose in the solution and, therefore, on the actual amount of nonbound water. Improved solubility of the more hydrophilic compounds due to their orientation toward the hydration shells of the sugar molecules was considered. The viscosity of nonassociated water forming the microregion for mass transfer of volatiles was considered instead of the bulk solution viscosity. Experimental validation of the model by real-time measurements of dynamic flavor release using foodlike flavor concentrations confirmed the above theory. Increasing sucrose concentrations resulted predominantly in increased flavor release, and bulk solution viscosity showed no effect. PMID- 12903971 TI - Comparison of natural and roasted Turkish tombul hazelnut (Corylus avellana L.) volatiles and flavor by DHA/GC/MS and descriptive sensory analysis. AB - Natural (raw) and roasted hazelnuts were compared for their differences in volatile components and sensory responses. A total of 79 compounds were detected in both hazelnuts, of which 39 (27 positive, 5 tentative, and 7 unknown) were detected in natural hazelnut and 71 (40 positive, 14 tentative, and 17 unknown) were detected in roasted hazelnut. These included ketones, aldehydes, pyrazines, alcohols, aromatic hydrocarbons, furans, pyrroles, terpenes, and acids. Pyrazines, pyrroles, terpenes, and acids were detected in roasted hazelnut only. Concentrations of several compounds increased as a result of roasting and these may play significant roles in the flavor of roasted hazelnut. Pyrazines together with ketones, aldehydes, furans, and pyrroles may contribute to the characteristic roasted aroma of hazelnut. Descriptive sensory analysis (DSA) showed that some flavor attributes such as "aftertaste", "burnt", "coffee/chocolate-like", "roasty", and "sweet" were rated significantly higher in roasted hazelnut compared to its natural counterpart. Natural and roasted hazelnuts can be distinguished using these attributes. PMID- 12903972 TI - Hydrolysis of wine aroma precursors during malolactic fermentation with four commercial starter cultures of Oenococcus oeni. AB - The ability of four commercial preparations of Oenococcus oeni lactic acid bacteria (EQ 54, Lalvin OSU, Uvaferm Alpha, and Lalvin 31) to hydrolyze wine aroma precursors was evaluated by measuring the concentration of free and bound aroma compounds at the end of malolactic fermentation carried out in model wines containing a mixture of glycosides extracted from Muscat wine. At pH 3.4 there was a decrease in glycosylated compounds matched by a concomitant increase in free forms in all starter cultures tested. When malolactic fermentation was carried out at pH 3.2, a significant decrease in the ability to hydrolyze aroma precursors was observed for two of the cultures tested (Uvaferm Alpha and Lalvin 31). Large differences in the extent of hydrolysis and in the specificity of this activity toward specific aroma precursors were observed and appeared to be related to the chemical structure of the aglycon as well as to individual characteristics of each starter culture. The amounts of glycosylated aroma compounds released during malolactic fermentation suggest that O. oeni can alter the sensory characteristics of wine through the hydrolysis of aroma precursors. PMID- 12903973 TI - Comparison of free amino acids profile in honey from three Argentinian regions. AB - The physical chemistry characteristics of honey are directly related to floral origin and, as a result, to the production region. There are some available methods that can determine the botanical or geographical origin of honey such as the free amino acids profile analysis. This paper reports data on the free amino acid composition, determined by reversed-phase high-performance liquid chromatography UV detection on 56 honey samples from three different Argentine regions, with characteristic apiarian flora. To evaluate if the quantified amino acid could be used to verify the geographical or botanical origin of honey, statistical analyses were performed. The cluster analysis showed that samples were grouped in clusters related to sampling regions and more strictly to apiarian flora around apiaries. Each cluster appears associated, in accordance with the principal component analysis, to high or low concentrations of different amino acids. PMID- 12903974 TI - Effects of non-covalent interactions with 5-O-caffeoylquinic acid (chlorogenic acid) on the heat denaturation and solubility of globular proteins. AB - The non-covalent interactions between the monomeric phenolic compound chlorogenic acid (5-CQA) and bovine serum albumin (BSA), lysozyme, and alpha-lactalbumin were characterized, and their effect on protein properties was examined. 5-CQA had a low affinity for all three proteins, and these interactions seemed to show a negative cooperativity. 5-CQA-BSA binding decreased with increasing temperature, whereas pH (pH 3.0 compared to pH 7.0) and ionic strength had no pronounced effect. At high 5-CQA/protein molar ratios, both the denaturation enthalpy and temperature of BSA increased; however, covalent bonds were created at high temperatures. The presence of 5-CQA had no effect on the solubility of BSA and alpha-lactalbumin as a function of pH, whereas it decreased lysozyme solubility at alkaline pH due to covalent interactions. These results indicate that the non covalent interactions with 5-CQA do not have pronounced effects on the functional properties of globular proteins in food systems. PMID- 12903975 TI - Isolation and thermal characterization of an acidic isoperoxidase from turnip roots. AB - An acidic peroxidase (pI approximately 2.5) was purified from turnip roots (TAP), and its thermal properties were evaluated. TAP is a monomeric protein having a molecular weight (MW) of 49 kDa and a carbohydrate content accounting for 18% of the MW. The yield of pure TAP was relatively high ( approximately 2 mg/kg of fresh roots), with a specific activity of 1810 2,2'-azinobis(3 ethylbenzthiazoline-6-sulfonic acid) (ABTS) units/mg at pH 6. The activity increased 4-fold at the optimum pH (4.0) to 7250 ABTS units/mg, higher than that of most peroxidases. TAP was heat stable; heat treatment of 25 min at 60 degrees C resulted in 90% initial activity retention, whereas an activity of 20% was retained after 25 min of heating at 80 degrees C. TAP regained 85% of its original activity within 90 min of incubation at 25 degrees C, following heat treatment at 70 degrees C for 25 min. Thermal inactivation caused noticeable changes in the heme environment as evaluated by circular dichroism and visible spectrophotometry. TAP was rapidly denatured by heating in the presence of 1.0 mM ethylene glycol bis(beta-aminoethyl ether) N,N,N',N'-tetraacetic acid, but the Soret band and activity were fully recovered by adding an excess of Ca(2+). This is further evidence that Ca(2+) plays an important role in the stability of TAP. The high specific activity of TAP, together with its relatively high thermal stability, has high potential for applications in which a thermally stable enzyme is required. PMID- 12903976 TI - Effect of low and high pH treatment on the functional properties of cod muscle proteins. AB - The functional properties of cod myosin and washed cod mince (myofibrillar protein fraction) treated at high (11) and low (2.5) pH were investigated after pH readjustment to 7.5. The solubility of refolded myosin was essentially the same as the native myosin. The pH-treated myofibrillar proteins had increased solubility over the whole ionic strength range studied. Acid and alkali treatment gave myosin and myofibrillar proteins improved emulsification properties, which were correlated with an increase in surface hydrophobicity and surface/interfacial activity. Enhanced gel strength was observed with acid- and alkali-treated myosin compared to native myosin, while the same treatment did not significantly improve the gel strength of acid- and alkali-treated myofibrillar proteins. The acid- and alkali-treated protein samples unfolded and gelled at a lower temperature than did the native proteins, suggesting a less conformationally stable structure of the refolded proteins. Functional studies show that acid and alkali treatment, which leads to partial unfolding of myosin may improve functional properties of cod myosin and myofibrillar proteins, with the greatest improvement being from the alkali treatment. The results also show that improvements in functionality were directly linked to the extent of partial unfolding of myosin on acid and alkali unfolding and refolding. PMID- 12903977 TI - Determination of high molecular mass polycyclic aromatic hydrocarbons in a typical Italian smoked cheese by HPLC-FL. AB - High-performance liquid chromatography with fluorescence detection was used for the analysis of polycyclic aromatic hydrocarbons (PAHs), in "Diavoletto" smoked cheese. Such cheese is typically produced in the Sorrento peninsula, and it is smoked commonly with different materials of vegetable origin. The importance of the smoking generation material is proven by the attention that the EU is paying in indicating the list of wood that may be used to produce smoking flavor agents. The PAHs considered are classified as "probable human carcinogens" by the U.S. Environmental Protection Agency for sufficient data from animal bioassays. The smoked samples contained high molecular mass PAHs with different levels ranging from 0.12 to 6.21 microg/kg. The determination was carried out also on liquid smoking flavor agents, smoke-flavored cheese, and nonsmoked cheese to measure the level of contamination before the treatment. PMID- 12903978 TI - Studies of selenium-containing volatiles in roasted coffee. AB - Coffee has been an important and heavily used beverage in many cultures over a long period of time. Although sulfur species have been found to be abundant constituents, no work to date has explored the presence of selenium analogues. Investigation of volatile selenium species from green coffee beans, roasted beans, and brewed coffee drink was performed using solid phase microextraction (SPME) sample preconcentration in conjunction with GC/ICP-MS. Several volatile selenium species at trace levels were detected from roasted coffee beans as well as in the steam from brewed coffee drinks. No detectable selenium (and sulfur) species, however, were found in the headspace of green beans, indicating that selenium-containing volatiles are formed during roasting, as is the case for the sulfur volatiles. Matching standards were prepared and used to identify the compounds found in coffee. Artificial supplementation of the green coffee beans with selenium before roasting was performed to further characterize the selenium containing volatiles formed during the coffee-roasting process. PMID- 12903979 TI - Extraction of carotenoids from feces, enabling the bioavailability of beta carotene to be studied in Indonesian children. AB - Previously, we have presented a method for quantifying beta-carotene bioavailability based on analysis in serum, following administration of (13)C labeled beta-carotene. Because stool samples can be collected noninvasively, we have now extended the method to measure the bioavailability based on measurements in feces. An extraction method was developed to enable measurement of concentrations and degree of isotopic enrichment of retinol, retinyl palmitate and carotenoids in feces. Relative bioavailability of beta-carotene from pumpkin (n = 6) was found to be 1.8 times (interval, 0.6, 5.5) greater than that from spinach (n = 8), based on data from feces compared with 1.7 times (interval, 0.9, 3.1) based on data from serum. PMID- 12903980 TI - Prolonged transit time through the stomach and small intestine improves iron dialyzability and uptake in vitro. AB - The iron dialyzability and uptake in relation to transit time through the stomach and small intestine was investigated using a dynamic in vitro gastrointestinal model in combination with Caco-2 cells. Three test meals were evaluated, consisting of lactic fermented vegetables with white (I) or whole meal bread (II) and of sourdough-fermented rye bread (III). Three transit times were tested (fast, medium, and slow transport). Iron dialyzability and absorption differed significantly between medium and slow transit time for meal I and between fast and medium transit time for meal III. For meal II, high in phytate, the iron dialyzability and absorption were low irrespective of transit time. The meals could be ranked with respect to iron dialyzability and uptake in the order I > III > II. Although the in vitro models used have limitations compared to in vivo experiments, the results suggest that an increased transit time may improve iron availability. PMID- 12903981 TI - Iron and zinc absorption from two bean (Phaseolus vulgaris L.) genotypes in young women. AB - Extrinsic and intrinsic iron and zinc labels were used to test iron and zinc absorption from two bean (Phaseolus vulgaris) genotypes, containing normal (common beans, CB) or higher (HFeZnB) iron and zinc concentrations, fed in single meals to young women with low iron reserves. The women were divided into two groups, with one receiving a CB test meal (n = 12) and the other, an HFeZnB test meal (n = 11). The beans were intrinsically labeled hydroponically with (55)Fe (CB and HFeZnB) and with (70)Zn (HFeZnB). Concentrations of zinc and iron were 98 and 65% higher, respectively, in HFeZnB as compared to CB, but phytic acid contents were similar. Extrinsic labels were (59)Fe (CB and HFeZnB), (67)Zn (CB), and (68)Zn (HFeZnB). Iron and zinc percent absorption levels were calculated from radio-iron activity in red blood cells and from urinary excretion of zinc isotopes. Intrinsic and extrinsic iron absorption measures were highly correlated (R (2) = 0.986) (average extrinsic/intrinsic ratio was 1.00). Iron absorption was low (geometric mean < 2%) in both bean types, and total iron absorbed was not different between types. Intrinsic zinc absorption from the HFeZn beans was higher than extrinsic absorption (15.2% vs 13.4%, p < 0.05) (average extrinsic/intrinsic was 0.90). The correlation between intrinsic and extrinsic zinc measures was not as high as that for iron (R (2) = 0.719). Percent zinc absorption levels were similar in both bean types, but total extrinsic zinc absorbed was 90% higher (p < 0.05) from the HFeZnB meal. Thus, the less expensive and time-consuming extrinsic labeling may be used to screen various varieties of beans for iron bioavailability in humans, but it underestimates zinc absorption by approximately 10%. Selective breeding for high-zinc bean genotypes may improve zinc status. However, high-iron genotypes appear to have little effect on iron status when fed alone in single meals to women with low iron reserves. PMID- 12903982 TI - Effect of natural and controlled fermentation on chemical composition and nutrient dialyzability from beans (Phaseolus vulgaris L.). AB - The effect of natural and controlled fermentation with an inoculum of Lactobacillus plantarum and additional thermal treatment (dry heat at 120 degrees C for 20 min) on the availability of N, P, Fe, Cu, Zn, Ca, and Mg from Phaseolus vulgaris L. var. carrilla was estimated using an in vitro method based on equilibrium dialysis. Natural and controlled fermentations caused significant reductions in the pH and phytate content (36%) of the bean flours, with a concomitant increase in the titratable acidity and free phosphorus content, and had no effect on the other nutrients studied. The percentage of dialyzable N, P, Cu, and Mg was significantly improved by both types of fermentation, whereas Zn dialyzability was significantly reduced. The greatest reduction was observed for the bean flour fermented with an inoculum of L. plantarum. The percentage of dialyzable Fe improved significantly as a result of natural fermentation but was not affected by controlled fermentation. The application of dry heat at 120 degrees C for 20 min caused a significant increase in Fe dialyzability and a further reduction in the percentage of dialyzable Zn in fermented bean flours but did not affect the dialyzability of the other nutrients studied. PMID- 12903983 TI - Inhibitory effects of (-)-epigallocatechin gallate on the mutation, DNA strand cleavage, and DNA adduct formation by heterocyclic amines. AB - Green tea is known to be a potential chemopreventive agent against cancer. In this study, we investigated the inhibitory activities of tea extracts, and in particular the polyphenolic component (-)-epigallocatechin gallate (EGCG), against heterocyclic amine-induced genotoxicity. The tea extracts displayed inhibition of 2-hydroxyamino-6-methyldipyrido[1,2-a,3',2'-d]imidazole (Glu-P 1(NHOH))-induced mutagenicity. This inhibition can be accounted for by the presence of EGCG in the extracts. The mutagenic effect of Glu-P-1(NHOH), which induces single-strand cleavage in supercoiled circular DNA under neutral conditions, was inhibited by EGCG. Using the Drosophila repair test, a test for gross DNA damage, and DNA adduct detection by (32)P-postlabeling, we showed that EGCG prevented 2-amino-3,8-dimethylimidazo[4,5-f]quinoline-induced DNA damage and adduct formation in insect DNA. EGCG was found to accelerate the degradation of Glu-P-1(NHOH) in vitro. This observation suggested that the inhibition by EGCG is associated with an accelerated degradation of metabolically activated heterocyclic amines. PMID- 12903984 TI - Preparation of antimicrobial reduced lysozyme compatible in food applications. AB - The structural and antimicrobial functions of lysozyme reduced with food compatible reducing agents-cysteine (Cys) and glutathione (GSH)-were investigated. The disulfide bonds were partially reduced by thiol-disulfide exchange reactions under heat-induced denaturing conditions from 55 to 90 degrees C. The results showed that treatment of lysozyme with Cys and GSH resulted in the introduction of new half-cystine residues (2-3 residues/mol of protein). The released SH groups, in turn, rendered the lysozyme molecule more flexible, being accompanied by a dramatic increase in the surface hydrophobicity and exposure of tryptophan residues. As a consequence, the resulting reduced lysozymes were more capable of binding to lipopolysaccharides (LPS) and permeabilizing the bacterial outer membrane, as evidenced by the liposome leakage experiment, than were native or heated lysozyme. Both reduced lysozymes displayed significantly higher antimicrobial activity than native or heated lysozyme against Salmonella enteritidis (SE) in sodium phosphate buffer (10 mM, pH 7.2) at 30 degrees C for 1 h. Their minimal inhibitory concentrations (MICs) against the tested bacteria were about 150- and 25-fold lower than their respective MICs of native or heated lysozyme. The results suggest that partially reduced lysozyme could be used as a potential antimicrobial agent for prevention of SE attack. PMID- 12903986 TI - Influence of roasting levels on ochratoxin a content in coffee. AB - Because of inconsistent and contradictory results from investigations concerning the influence of roasting process on the ochratoxin A content in coffee beans, a study was undertaken to assess the elimination of ochratoxin A during the roasting process. Four different green coffee samples, naturally contaminated with ochratoxin A, were submitted to different roasting conditions (light, medium, and dark) and analyzed for roasting parameters (weight loss, color change, density, and moisture content) and ochratoxin A content. The ochratoxin A content of green coffee was reduced by the roasting process; in particular, consistently high percentages of ochratoxin A reduction were found in the highest contaminated samples. This reduction was influenced by the severity of the thermal process and was generally related to the initial ochratoxin A content. Samples obtained with roasting parameters suitable for a typical Italian espresso coffee brew showed reductions of >90% in the ochratoxin A content, in both high and low contaminated samples. Moreover, the presence of off-flavors and visual defects was not found to be directly related to the ochratoxin A content in the green coffee samples. PMID- 12903985 TI - Effect of stevioside and steviol on the developing broiler embryos. AB - At day 7 of incubation, fertile broiler eggs were injected with different amounts of stevioside and steviol of 0.08, 0.8, or 4 mg stevioside/egg and 0.025, 0.25, or 1.25 mg steviol/egg. At hatch (day 21) and 1 week later, not any influence of the different treatments could be found on embryonic mortality, body weight of the hatchlings, deformations (e.g., bone, beak, and head malformations, abnormal feathering, open vent), or abnormal development of the gonads. No stevioside or steviol could be detected in the blood of the hatchlings. The hatchlings developed normally. It is concluded that prenatal exposure to stevioside and steviol is not toxic for the chicken embryo. PMID- 12903987 TI - Deoxynivalenol removal from barley intended as swine feed through the use of an abrasive pearling procedure. AB - Samples of naturally contaminated hulled barley, with varying deoxynivalenol concentrations, were subjected to an abrasive type dehulling procedure. The remaining grain fractions were analyzed for weight remaining (%), deoxynivalenol (ppm), crude protein (%CP), neutral detergent fiber (%NDF), ash (%ASH), gross energy (GE; kcal/kg), and calculated digestible energy values (DE; kcal/kg). Following the initial 15 s of pearling, 85% of the grain mass remained. Additional pearling resulted in a linear decline of grain mass. Following 15 s of pearling, the grain contained 34% of the initial deoxynivalenol content, irrespective of the initial level of contamination. Further pearling resulted in continued significant (p < 0.05) reductions in the percent of deoxynivalenol remaining to a level of 7.9% after 120 s but with significant losses in grain mass. Pearling can serve as an effective means of reducing the deoxynivalenol content of barley, with improvements in nutrient levels. However, the need to reduce the deoxynivalenol content of contaminated barley to less than 1 ppm for swine will necessitate the removal of a significant amount of the grain mass for heavily contaminated samples. PMID- 12903988 TI - Safety evaluation of organoarsenical species in edible Porphyra from the China Sea. AB - A study was carried out to determine arsenic species in Porphyra seaweed originating from the China Sea. Information about arsenic species in Porphyra was provided by HPLC-ICP-MS and ES-MS-MS. The total arsenic concentrations of Porphyra samples from five different producing areas ranged from 2.1 to 21.6 mg/kg. The analysis report also showed that arsenosugars were the only arsenic species that could be detected in all of the extracts of samples. Arsenosugar PO(4) was the major compound in most samples (0.3-13.9 mg/kg of dry weight), followed by arsenosugar OH (0.7-6.2 mg/kg of dry weight). A further experiment was done to investigate the stability of arsenosugars in the process of being heated. It was observed that the arsenosugars were stable during a short-term heating at 100 degrees C. Their stability in human ingestion was also studied. A substantial increase of dimethylarsinic acid (DMA) was detected in urine samples collected from six volunteers after the consumption of this seaweed. The results obtained indicated that arsenosugars had been metabolized to DMA, which is more toxic than arsenosugars. From this point of view, consumers should consider the possible adverse effects of edible Porphyra on human health and choose those Porphyra having lower arsenic concentrations. PMID- 12903989 TI - Comment on digestibility of food allergens and nonallergenic proteins in simulated gastric fluid and simulated intestinal fluid--a comparative study. PMID- 12903992 TI - Application of host-guest chemistry in nanotube-based device fabrication: photochemically controlled immobilization of azobenzene nanotubes on patterned alpha-CD monolayer/Au substrates via molecular recognition. AB - Azobenzene-functionalized nanotubes recognized and attached onto well-defined complementary regions of thiolated alpha-CD SAM/Au substrates via host-guest molecular recognition. The binding between the azobenzene nanotubes and the alpha CD SAM/Au substrates was controlled by UV irradiation. The light-induced attachment-detachment of the azobenzene nanotubes on the alpha-CD SAMs was reversible. Some of the nanotubes were capable of interconnecting two Au substrates. This smart building block may be applied to build photoactive nanometer-sized mechanical switches in electronics. PMID- 12903993 TI - Asymmetric addition of alkylzinc reagents to cyclic alpha,beta-unsaturated ketones and a tandem enantioselective addition/diastereoselective epoxidation with dioxygen. AB - Although over 100 catalysts have been reported to catalyze the asymmetric addition of alkyl groups to aldehydes, these catalysts fail to promote additions to ketones with >90% enantioselectivity. This paper describes the asymmetric 1,2 addition of alkyl groups to conjugated cyclic enones to give allylic alcohols with chiral quaternary centers. The resultant allylic alcohols are converted into epoxy alcohols with excellent diastereoselectivities. Treatment of the epoxy alcohols with BF3.OEt2 induces a semipinacol rearrangement to provide alpha,alpha dialkyl-beta-hydroxy ketones with all-carbon chiral quaternary centers. We also report a one-pot procedure for the asymmetric addition/diastereoselective epoxidation reaction. Simply exposing the reaction mixture to dioxygen after the asymmetric addition reaction is complete results in epoxidation of the allylic alcohol with high diastereoselectivity. PMID- 12903994 TI - A reversible tube-to-rod transition in a block copolymer micelle. AB - A remarkable morphology transition occurs with a change in temperature for a diblock copolymer [poly(ferrocenyldimethylsilane-b-dimethylsiloxane) (PFS40-b PDMS480, PDI = 1.01)] in n-decane solution. This polymer, which forms nanotubes at 25 degrees C, rearranges to form short dense rods when the solution is heated to 50 degrees C. When the solution is cooled to 25 degrees C, the system evolves back to nanotubes. These experiments demonstrate that both structures are dynamic and represent equilibrium states of the material. Contrast matching static light scattering measurements on the short dense rods show that the insoluble PFS core is rigid and has a length distribution similar to that seen in electron microscopy images. PMID- 12903995 TI - A novel kinetic trap for NO release from cytochrome c': a possible mechanism for NO release from activated soluble guanylate cyclase. AB - Flash photolysis studies on the five-coordinate heme nitrosyl of Alcaligenes xylosoxidans cytochrome c' were carried out to investigate the ramifications of its proximal nitrosyl ligand on NO release. Delta absorbance spectra recorded 5 ms after photolysis indicate that approximately 5% of the photolyzed hemes are converted to a five-coordinate high spin ferrous state, revealing that reattachment of the endogenous His ligand is fast enough to trap some of the photolyzed heme. Analysis of NO rebinding suggests that the photolyzed ferrous protein is initially in a strained conformation, which relaxes on a millisecond time scale. The strained ferrous heme appears to contain a significantly labilized Fe-His bond, which allows direct second-order rebinding to the proximal face at high NO-concentrations. In contrast, the NO-binding properties of the relaxed conformation are similar to those previously observed in stopped-flow studies, which showed that a five-coordinate heme-nitrosyl is formed via a six coordinate intermediate. The discovery of a rapid proximal His ligand reattachment to NO-dissociated heme reveals a novel "kinetic trap" mechanism for lowering the five-coordinate heme nitrosyl population in response to decreased ambient NO concentrations. Thus, NO dissociation from the five-coordinate heme nitrosyl, whether thermal or photochemical, is followed by rapid, and only slowly reversible, His reattachment which acts to kinetically trap the heme in its five coordinate ferrous state. Because return to the five-coordinate heme nitrosyl requires two NO-dependent steps, the protein uses a kind of kinetic amplification of the thermodynamic dissociation that occurs in response to decreased NO concentrations. The implications of this "kinetic-trap" mechanism for NO release from soluble guanylate cyclase are discussed. PMID- 12903996 TI - Mixed-element capture agents: a simple strategy for the construction of synthetic, high-affinity protein capture ligands. AB - Demonstration of a simple strategy to generate synthetic high-affinity protein capture agents of practical utility for protein-detecting microarrays. The model study highlights capture of the MBP-Mdm2 fusion protein on a solid support by a linear sequence of peptides that bind to the two individual polypeptide chains. PMID- 12903997 TI - Spontaneous catalytic generation of nitric oxide from S-nitrosothiols at the surface of polymer films doped with lipophilic copperII complex. AB - A new approach for preparing potentially more blood-compatible nitric oxide (NO) generating polymeric materials is described. The method involves creating polymeric films that have catalytic sites within (lipophilic copper(II) complex) that are capable of converting endogenous S-nitrosothiols present in blood (S nitrosoglutathione (GSNO), S-nitrosocysteine (CysNO), etc.) to NO. The catalytic NO generation reaction involves the initial reduction of Cu(II) to Cu(I) within the complex by appropriate reducing agents (e.g., thiolates or ascorbate), followed by the reduction of S-nitrosothiols to NO by the Cu(I) complex at the polymer/solution interface. The NO fluxes observed when PVC or polyurethane films containing the copper(II) complex are placed in solutions containing physiological levels of nitrosothiols (muM levels) reach ca. 8 x 10-10 mol cm-2 min-1, greater than that produced by normal endothelial cells that line all healthy blood vessels. It is thus anticipated that this spontaneous catalytic generation of NO from endogenous nitrosothiols will render such polymeric materials more thromboresistant when in contact with blood in vivo. PMID- 12903998 TI - Biomimetic synthesis of (+/-)-pinnatal and (+/-)-sterekunthal A. AB - Concise biomimetic syntheses of the antimalarial naphthoquinones (+/-)-pinnatal and (+/-)-sterekunthal A are described. PMID- 12903999 TI - Calculation of dipole-shielding polarizabilities (sigma(alphabetagamma)I): the influence of uniform electric field effects on the shielding of backbone nuclei in proteins. AB - A significant contribution to the chemical shielding of a nucleus can arise from uniform electric fields that act to distort the electronic charge distribution surrounding a nucleus and, hence, affect the nuclear shielding. It has been shown by Buckingham (Buckingham, A. D. Can. J. Chem. 1960, 38, 300) that the nuclear magnetic shielding tensor sigmaalphabetaI of a nucleus in the presence of an external weak static uniform electric field E may be expanded using sigmaalphabetaI(E) = sigmaalphabetaI + sigmaalphabetagammaIEgamma + 1/2sigmaalphabetagammadeltaIEgammaEdelta + sigmaalphabeta,gammadeltaIEgammadelta ... The third rank tensor sigmaalphabetagammaI is referred to as the dipole shielding polarizability and describes the nonlinear response of the electron cloud to first order in E, muI, and B0. We report calculations of sigmaalphabetagammaI for the N, HN, and C' nuclei in N-methyl acetamide (NMA) and show that these tensors can be used to provide considerable insight into the behavior of uniform electric fields upon the shielding of backbone nuclei in proteins. The sigmaalphabetagammaI values for the N, HN, and C' of NMA were calculated using the continuous transformation of the origin of the current density (CTOCD) scheme with the diamagnetic contribution set to zero (CTOCD-DZ). Values are given for the individual tensor components of sigmaalphabetagammaI for each nucleus. To test that the calculations have provided a reasonable estimate for the sigmaalphabetagammaI of N, HN, and C' nuclei in proteins, a pH titration was performed using Hen Lysozyme (HEWL). The pH-induced isotropic shielding changes for the C', N, and HN nuclei in some peptide bonds close to E35 ( approximately <8 A) were extracted from sets of fitted titration curves. Assuming the experimental shielding changes arise solely from uniform electric field effects caused by the deprotonation of E35, without any other pH-induced structural alterations which might lead to a shielding change, the experimental shielding differences were compared to those calculated via the product Agamma(I).Egamma where Agamma(I) = (1/3)sigmaalphaalphagammaIota. The agreement with the experimental data is in many cases reasonable and suggests that, within the Buckingham formalism, the complete sigmaalphabetagammaI tensors reported here will be helpful to resolve the importance of uniform electric fields upon isotropic and anisotropic shielding in proteins and their complexes. PMID- 12904000 TI - Ketonization of incarcerated acetophenone enol. AB - A free-standing simple enol has been generated inside a carceplex. Rates of ketonization under various conditions were determined; ketonization is extremely slow as compared to the rate in solution. Complexed water is required, and the mechanism proceeds via formation of an enolate followed by protonation at carbon by the same molecule of water that removed the proton from the enol. Acid or base retards ketonization by removing water from the cavity. PMID- 12904001 TI - Chelating bis(thiophosphinic amidate)s as versatile supporting ligands for the group 3 metals. An application to the synthesis of highly active catalysts for intramolecular alkene hydroamination. AB - Bis(thiophosphinic amidate) complexes (i.e., 1) of representative group 3 and lanthanide metals have been quantiatively prepared in situ from the corresponding thiophosphinic amides and Ln[N(TMS)2]3. These unusual pentacoordinate complexes exhibit very high activity as catalysts for intramolecular alkene hydroamination. PMID- 12904002 TI - Ultrafast fluorescence investigation of excitation energy transfer in different dendritic core branched structures. AB - The mechanism of energy transport in branching structures is suggestively related to the geometry of the multichromophore architecture. In organic conjugated dendrimers, both incoherent (hopping) and coherent energy transfer processes have been observed from different dendritic architectures with different building blocks. In this communication, we report the investigation of three fundamental dendritic architectures (G0) with the same attached chromophores, but with different core atoms, C, N, and P. The synthesis of a phosphorus-containing G0 system with distyrylbenzene chromophores is provided. These three systems provide a comparison by which the relative interaction of branching chromophores can be compared on the basis of their different branching centers. Ultrafast fluorescence anisotropy measurements provide a dual measure of the geometry of the chromophores around the different central units as well as the strength of the interactions among chromophores. The nitrogen-cored system appeared to have both the strongest coupling of chromophore excitation as well as the most planar geometry of the three. Interestingly, the phosphorus system appeared to have the least planar geometry, and its interaction strength was found to be stronger than that observed for the carbon system. These results provide a comparison of the energy migration dynamics of the most common and new dendritic architectures with applications for light emission and light harvesting. PMID- 12904003 TI - Hydrocarbation chemistry proceeding from nickel carbenes. AB - Nickel carbene complex 2 [Ni(triphos)C(H)N(H)xylyl]2+(BF4-)2 reacts with alkenes quantitatively and regiospecifically to give the anti-Markovnikov hydrocarbation products. X-ray crystallography shows significant iminium alkyl character of the hydrocarbation products, similar to that observed in parent carbene 2. Mechanistic studies suggest the importance of a "hydride" pathway over "alkene" (metallocycle formation or carbocation) pathways. PMID- 12904004 TI - Observation of individual transitions in magnetically equivalent spin systems. AB - Individual transitions of magnetically equivalent spin systems such as methyl groups residing on isotropically tumbling molecules in solution usually cannot be observed as multiplet-split NMR lines. We propose a pair of NMR experiments, 2D [13C,1Halphaalpha]Methyl and [13C,1Hbetabeta]Methyl HSQC, to overcome this limitation and enable direct and selective observation of individual 1H transitions in 13C-labeled methyl spin systems. Immediate applications include quantitative measurements of 1H-1H residual dipolar couplings (RDC) and cross correlated relaxation between 1H chemical shift anisotropy and 1H-1H dipole dipole interactions. The use of the experiments for the measurement of RDCs is demonstrated with two proteins, one weakly aligned by means of Pf1 phages and the other using a naturally present paramagnetic heme group. PMID- 12904005 TI - Unprecedented double C-C bond cleavage of a cyclopentadienyl ligand. AB - Double C-C bond cleavage of a cyclopentadienyl ligand proceeded to titanacyclopentadienes when 2 equiv of nitriles were added and the resulting two carbon unit and three-carbon unit were converted into a benzene derivative and a pyridine derivative, respectively, in one-pot. PMID- 12904006 TI - Enantioselective hydrogenation of tetrasubstituted olefins of cyclic beta (acylamino)acrylates. AB - Hydrogenation of a series of cyclic beta-(acylamino)acrylates with tetrasubstituted olefins has been accomplished successfully with the use of Ru catalysts with chiral biaryl ligands such as C3-TunaPhos, and up to over 99% ee's have been achieved. This methodology provides an efficient catalytic method for the synthesis of both cis and trans chiral cyclic beta-amino acid derivatives. PMID- 12904007 TI - Light-induced aminocarbene to imine dyotropic rearrangement in a chromium(0) center: an unprecedented reaction pathway. AB - 1,2-Dyotropic rearrangement can be induced by irradiation of properly functionalized Fischer carbenes. This novel reaction takes place by a stepwise mechanism and with double inversion of configuration at the static scaffold. Good yields are obtained with both cyclic and acyclic structures, thus suggesting that this unprecedented transformation can be extended to other organometallic systems. PMID- 12904008 TI - Beta-elimination of a phosphonate group from an alkoxy radical: an intramolecular acylation approach using an acylphosphonate as a carbonyl group acceptor. AB - On the basis of facile beta-elimination of a phosphonate group from an alkoxy radical, intramolecular acylation reaction has been developed, in which an acylphosphonate is used as an excellent carbonyl group radical acceptor. PMID- 12904009 TI - Macrolactonization via Ti(IV)-mediated epoxy-acid coupling: a total synthesis of (-)-dactylolide [and zampanolide]. AB - A total synthesis of dactylolide (1) is described. The key feature involves the Ti(IV)-mediated coupling of structurally complex "Sharpless epoxides" and carboxylic acids in either an intramolecular (macrolactonization) or an intermolecular mode. Other notable aspects include a proton-catalyzed, cis selective construction of the 4-methylenetetrahydropyran ring; a selective oxidation of an allylic alcohol in the presence of a 1,2-diol by an oxoammonium ion; an efficient ring-closing metathesis reaction of an in situ (bis-TMS) protected alpha,omega-diene-vic-diol; and an aluminum-mediated aza-aldol reaction of a primary amide to 1 to construct the acyclic carbinolamide in zampanolide. PMID- 12904010 TI - Catalytic C-H bond functionalization with palladiumII: aerobic oxidative annulations of indoles. AB - A palladium-catalyzed aerobic oxidative annulation of indoles is described. We have demonstrated that a variety of factors influence these cyclizations, and in particular the electronic nature of the pyridine ligand is crucial. It is also remarkable that these oxidative cyclizations can proceed in good yield despite background oxidative decomposition pathways, testament to the facile nature with which molecular oxygen can serve as the direct oxidant for Pd(0). We have also shown that the mechanism most likely involves initial indole palladation (formal C-H bond activation) followed by migratory insertion and beta-hydrogen elimination. PMID- 12904011 TI - A designed beta-hairpin peptide for molecular recognition of ATP in water. AB - A designed 12-residue beta-hairpin peptide with a diagonal tryptophan (Trp) pair was shown to bind ATP in water through a combination of aromatic and electrostatic interactions. The affinity for ATP was 5800 M-1 (DeltaG approximately -5.0 kcal/mol), a remarkable affinity for a short, structured peptide in water, consisting of entirely natural amino acid residues. Proton NMR measurements indicate that the adenine ring of the nucleotide is intercalated between the diagonal tryptophans in the bound state. Delineation of the contributions to ATP binding to the hairpin suggest that aromatic interactions contribute approximately -1.8 kcal/mol, while individual electrostatic interactions involving the ATP phosphates and positively charged side chains of the hairpin contribute approximately -1 kcal/mol each. The designed beta-hairpin receptor presents a novel minimalist system to investigate the energetic contributions to protein-nucleic acid recognition through the surface of a beta sheet. PMID- 12904013 TI - A highly efficient three-component coupling of aldehyde, alkyne, and amines via C H activation catalyzed by gold in water. AB - A highly efficient three-component-coupling of aldehyde, alkyne, and amine (A3 Coupling) catalyzed by gold via C-H activation was developed in water. The reaction is general, and nearly quantitative yields of the corresponding propargylamines were obtained in most cases. Both aromatic and aliphatic aldehydes and amines can be used for the reaction. No additional cocatalyst or activator is required. A mechanistic rationale for the reaction has been proposed. PMID- 12904012 TI - Enyne metathesis for the formation of macrocyclic 1,3-dienes. AB - Macrocyclic 1,3-dienes of a variety of ring sizes are formed in good yield via enyne metathesis. Both endo- and exo-products are observed depending on the size of the macrocycle. In general, 10-membered rings and smaller give exo-products, while 12-membered rings and larger give endo-products. The endo/exo selectivity and the E/Z ratio of the diene products can be further controlled by the presence of ethylene in the reaction. PMID- 12904014 TI - Fast back electron transfer prevents guanine damage by photoexcited thionine bound to DNA. AB - The phenothiazinium dye thionine has a high excited state reduction potential and is quenched by guanine on the femtosecond time scale. Here, we show by gel electrophoresis that irradiation of thionine with 599 nm light in the presence of an oligonucleotide duplex does not produce permanent DNA damage. Upon photoexcitation of thionine weakly associated with guanosine-5'-monophosphate, the reduced protonated thionine radical and neutral guanine radical are detected by transient absorption spectroscopy, indicating that the quenching of thionine by guanine occurs via an electron-transfer mechanism. The observation of radical formation without permanent guanine damage indicates that fast back electron transfer plays a critical role in governing the yield of damage by DNA-binding molecules. PMID- 12904015 TI - A potent and highly selective inhibitor of human alpha-1,3-fucosyltransferase via click chemistry. AB - Potent inhibitors of fucosyltransferases, and glycosyltransferases in general, have been elusive due to the inherent barriers surrounding the family of glycosyltransfer reactions. The problems of weak substrate affinity and low catalytic proficiency of fucosyltransferase was offset by recruiting additional binding features, in this case hydrophobic interactions, to produce a high affinity inhibitor, 24, with Ki = 62 nM. The molecule was identified from a GDP triazole library of 85 compounds, which was produced by the Cu(I)-catalyzed [2 + 3] cycloaddition reaction between azide and acetylene reactants, followed by in situ screening without product isolation. PMID- 12904017 TI - Coulomb explosion upon electron attachment to a four-coordinate monoanionic metal complex. AB - Electron capture to monoanionic metal complexes in high-energy collisions with sodium vapor is shown to occur with the formation of dianions. In this way, we prepared the small dianions Cr(SCN)42-, Fe(CN)42-, Pt(NO2)22-, and Pt(C2O4)22- in the gas phase. The Cr(SCN)42- dianion Coulomb explodes into Cr(SCN)3- and SCN- with a release of kinetic energy (3.2 +/- 0.4 eV) into translational energy of the fragments. The scheme provides a way to study charge dissociation reactions of molecular dianions that are too short-lived to survive extraction from the ion source. PMID- 12904016 TI - Identification of Glu166 as the general base in the acylation reaction of class A beta-lactamases through QM/MM modeling. AB - Bacterial class A beta-lactamases are responsible for the most known resistance against beta-lactam antibiotics. With the continuing rise in antibiotic resistance, improved knowledge of the mechanisms of action of these enzymes is needed in the development of effective therapeutic agents and strategies. The mechanism of the deacylation step in class A beta-lactamases is well accepted. In contrast, the mechanism of the acylation step has been uncertain, with several conflicting proposals put forward. We have modeled the acylation step in a class A beta-lactamase, using a combined quantum mechanics/molecular mechanics approach. The results provide an atomic level description of the reaction and show that Glu166 acts as the general base in the reaction, deprotonating Ser70 via an intervening water molecule. Ser70 acts as the nucleophile for attack on the lactam ring in a concerted reaction. The results do not rule out the importance of Lys73 in catalysis, in agreement with experimental data. PMID- 12904018 TI - Enolization of acetone in superheated water detected via radical formation. AB - Muoniated free radicals have been detected in muon-irradiated aqueous solutions of acetone at high temperatures and pressures. At temperatures below 250 degrees C, the radical product is consistent with muonium addition to the keto form of acetone. However, at higher temperatures, a different radical was detected, which is attributed to muonium addition to the enol form. Muon hyperfine coupling constants have been determined for both radicals over a wide range of temperatures, significantly extending the range of conditions under which these radicals and the keto-enol equilibrium have been studied. PMID- 12904019 TI - Toward a versatile allylation reagent: practical, enantioselective allylation of acylhydrazones using strained silacycles. AB - A highly practical method for the enantioselective allylation of acylhydrazones has been developed. The previously reported strained silacycle reagent 1 reacts with a wide variety of acylhydrazones to give the hydrazide products with good enantioselectivity (83-89% ee, typically). It has been demonstrated that the products may be isolated without chromatography by recrystallization in >/=98% ee. PMID- 12904020 TI - Surface-enhanced Raman scattering in the ultraviolet spectral region: UV-SERS on rhodium and ruthenium electrodes. AB - We report the first observation of surface-enhanced Raman scattering (SERS) excited with ultraviolet (UV) light from transition metal electrodes. Adsorbed pyridine and SCN- on rough rhodium (Rh) and ruthenium (Ru) electrodes, respectively, have been studied using 325 nm laser excitation. In contrast, the best enhancers in the visible and near infrared, silver and gold, do not produce UV-SERS. The experimental data of UV-SERS are in agreement with our preliminary theoretical calculation based on the electromagnetic enhancement mechanism. The enhancement factor is about 2 orders of magnitude for the Rh and Ru electrodes when they are excited at 325 nm. PMID- 12904021 TI - Observation of the marcus inverted region of electron transfer reactions at a liquid/liquid interface. AB - Scanning electrochemical microscopy was used to probe the influence of a driving force on the heterogeneous electron transfer (ET) processes at the externally polarized water/1,2-dichloroethane interface. Being a part of the driving force, the Galvani potential difference at the interface, Deltaowphi, can be quantitatively controlled in a wide range, allowing the precise measurements of the rate constants of the ET reactions. Two opposite systems were chosen in this work. One was 5,10,15,20-tetraphenyl 21H,23H-porphyrin zinc (ZnPor, O)/Fe(CN)64- (W), and the other was TCNQ (O)/Fe(CN)63- (W). For both systems studied, the relations between the rate constant and the Deltaowphi were of parabolic shape; that is, the rate constants increased initially with the Deltaowphi until reaching a maximum and then decreased steadily as the Deltaowphi increased further. This is in accordance with the prediction of the Marcus theory. To our knowledge, this is the first report that the Marcus inverted region can be observed electrochemically at an unmodified liquid/liquid interface with only one redox couple at each phase. The effect of the concentrations of the organic supporting electrolyte has also been discussed in detail. PMID- 12904022 TI - A concise route to benzofused macrolactones via ynolides: cycloproparadicicol. AB - A new facile synthesis has been developed for nanomolar Hsp90 inhibitor, cycloproparadicicol (2). Our approach relied on cobalt-complexation promoted RCM, in combination with tandem Diels-Alder/retro-Diels-Alder reactions to assemble the resorcycinylic macrolactone. PMID- 12904023 TI - A pi-basic rhenium center that effects cyclohexene isomerization to a beta agostic carbene ligand. AB - The molecule (PNPR)Re(H)4 (PNPR = (R2PCH2SiMe2)2N, R = iPr or cyclohexyl) reacts at 20 degrees C with 2 mol of cyclohexene to form equimolar cyclohexane and (PNPR)Re(H)2[=C(CH2)5]. This product is characterized by 1H, 13C, and 31P NMR and by X-ray diffraction as having one CH2 hydrogen (from a carbon located beta to Re) donating to the metal ("agostic CH"). This interaction occurs in preference to PNPR amide nitrogen pi-donation. DFT calculations confirm this agostic interaction, and show that the (PNPR)Re(H)2 fragment indeed reverses the greater stability of free olefin vs free carbene. PMID- 12904024 TI - The largest protein observed to fold by two-state kinetic mechanism does not obey contact-order correlation. AB - To probe the folding-energy landscape for a very large protein, we used Borrelia burgdorferi VlsE as a model. VlsE is a single-domain, predominantly alpha-helical protein with 341 residues. Remarkably, time-resolved folding and unfolding processes for VlsE follow two-state behavior. VlsE is by far the largest protein characterized that folds by a two-state kinetic mechanism. Thus, the common rule of thumb, that proteins larger than 110 residues fold by complex, multistate kinetic mechanisms, must be used with caution. In contrast with smaller helical proteins, the folding speed in water for VlsE is slow (5 +/- 2 s-1, pH 7, 20 degrees C) and does not agree (by 4 orders of magnitude in different directions) with the speeds predicted on the basis of native-state contact order and the topomer-search model. It is therefore questionable if the barrier height for folding is defined by gross topology for large two-state folders. PMID- 12904025 TI - In situ functionalization of porous silicon during the electrochemical formation process in ethanoic hydrofluoric acid solution. AB - In this work, the results of a new method for the preparation of porous silicon (PS) layers with in situ simultaneous functionalization with organic molecules are reported. The molecules of interest are dissolved in the HF ethanoic solution used to prepare the PS layers by partial anodic dissolution of a Si electrode. The method has been proved to be effective with various molecules. In this Communication, the case of PS functionalization with heptyne molecules, studied by FTIR spectroscopy, is reported in detail. The results demonstrate that this new functionalization method, accompanied by a low-level oxidation, is simple, fast, and effective and that it can allow the confinement of the adsorbed molecules selectively in a single layer of a PS stack. PMID- 12904026 TI - A new Co(2)(CO8-mediated tandem [5 + 1]/[2 + 2 + 1]-cycloaddition reaction: a one pot synthesis of tricyclic delta-lactones from cis-epoxy ene-ynes. AB - In the presence of Co2(CO)8 and CO, cis-epoxyalkynes bearing a tether olefin undergo a tandem [5 + 1]/[2 + 2 + 1]-cycloaddition to give tricyclic delta lactones efficiently in a one-pot operation. The reaction mechanism is proposed to involve a cobalt-coordinated cyclic allene species. PMID- 12904027 TI - Structural aspects for evolution of beta-lactamases from penicillin-binding proteins. AB - Penicillin-binding proteins (PBPs), biosynthetic enzymes of bacterial cell wall assembly, and beta-lactamases, resistance enzymes to beta-lactam antibiotics, are related to each other from an evolutionary point of view. Massova and Mobashery (Antimicrob. Agents Chemother. 1998, 42, 1-17) have proposed that for beta lactamases to have become effective at their function as antibiotic resistance enzymes, they would have had to undergo structure alterations such that they would not interact with the peptidoglycan, which is the substrate for PBPs. A cephalosporin analogue, 7beta-[N-Acetyl-L-alanyl-gamma-D-glutamyl-L-lysine]-3 acetoxymethyl-3-cephem-carboxylic acid (compound 6), was conceived and synthesized to test this notion. The X-ray structure of the complex of this cephalosporin bound to the active site of the deacylation-deficient Q120L/Y150E variant of the class C AmpC beta-lactamase from Escherichia coli was solved at 1.71 A resolution. This complex revealed that the surface for interaction with the strand of peptidoglycan that acylates the active site, which is present in PBPs, is absent in the -lactamase active site. Furthermore, insertion of a peptide in the beta-lactamase active site at a location where the second strand of peptidoglycan in some PBPs binds has effectively abolished the possibility for such interaction with the beta-lactamase. A 2.6 ns dynamics simulation was carried out for the complex, which revealed that the peptidoglycan surrogate (i.e., the active-site-bound ligand) undergoes substantial motion and is not stabilized for binding within the active site. These factors taken together disclose the set of structure modifications in the antibiotic resistance enzyme that prevent it from interacting with the peptidoglycan, en route to achieving catalytic proficiency for their intended function. PMID- 12904028 TI - pH as a trigger of peptide beta-sheet self-assembly and reversible switching between nematic and isotropic phases. AB - The hierarchical self-assembly of rationally designed synthetic peptides into beta-sheet tapes, ribbons, fibrils, and fibers opens up potentially useful routes to soft-solidlike materials such as hydrogels, organogels, or liquid crystals. Here, it is shown how incorporation of Glu (-CH(2)CH(2)COOH) or Orn ( CH(2)CH(2)CH(2)NH(2)) into the primary structure of an 11 amino acid peptide enables self-assembly to be rapidly (seconds) and reversibly controlled by simply changing pH. Solutions of monomeric peptide, typically at concentrations in excess of 0.003 v/v, can be switched within seconds to, for example, nematic gel states comprised of interconnected orientationally ordered arrays of fibrils or vice versa. This is to be compared with the lyophilized peptide dissolution route to nematic fluids and gels which is impracticably long, taking many hours or even days. An important design principle, that stabilization of fibrillar dispersions requires of the order of one unit of net positive or negative charge per peptide molecule, is first demonstrated and then used to design an 11 amino acid peptide P(11)-3 (CH(3)CO-Gln-Gln-Arg-Phe-Gln-Trp-Gln-Phe-Gln-Gln-Gln-NH(2)) whose self assembly behavior is independent of pH (1 < pH < 10). pH control is then incorporated by appropriately positioning Glu or Orn side chains so that the peptide-peptide free energy of interaction in the tapelike substructure is strongly influenced by direct electrostatic forces between gamma-COO(-) in Glu(-) or delta-NH(3)(+) in Orn(+), respectively. This design principle is illustrated by the behavior of two peptides: P(11)-4 (CH(3)CO-Gln-Gln-Arg-Phe-Glu-Trp-Glu-Phe Glu-Gln-Gln-NH(2)) which can be switched from its nematic to its isotropic fluid state by increasing pH and P(11)-5 (CH(3)CO-Gln-Gln-Orn-Phe-Orn-Trp-Orn-Phe-Gln Gln-Gln-NH(2)) designed to exhibit the converse behavior. Acid-base titrations of fibrillar dispersions reveal deprotonation of the gamma-COOH of Glu or of the delta-NH(3)(+) of Orn(+) occurs over wide bands of up to 5 pH units, a feature of polyelectrolytes. The values of the energy parameters controlling self-assembly can therefore be smoothly and continuously varied by changing pH. This enables isotropic fluid-to-nematic transitions to be triggered by relatively small additions of acid or base, typically 1 part in 10(3) by volume of 1 M HCl or NaOH. PMID- 12904029 TI - Unprecedented monofunctional metalation of adenine nucleobase in guanine- and thymine-containing dinucleotide sequences by a cytotoxic platinum-acridine hybrid agent. AB - We have investigated the reactions of [PtCl(en)(ACRAMTU-S)](NO(3))(2) (2) (en = ethane-1,2-diamine; ACRAMTU = 1-[2-(acridin-9-ylamino)ethyl]-1,3 dimethylthiourea, acridinium cation, 1), the prototype of a new class of cytotoxic DNA-targeted agents, with 2'-deoxyguanosine (dGuo) and random-sequence native DNA by in-line liquid chromatography/mass spectrometry (LC/MS) and NMR spectroscopy ((1)H, (195)Pt) to identify the covalent adducts formed by platinum. In the mononucleoside model system, two adducts are observed, [Pt(en)(ACRAMTU)(dGuo)](3+) (P1, major) and [Pt(en)(dGuo)(2)](2+) (P2, minor). The reaction, which proceeds significantly slower (half-life 11-12 h at 37 degrees C, pH 6.5) than analogous reactions with cisplatin and reactions of 2 with double-stranded DNA, results in the unexpected displacement of the sulfur bound acridine ligand in approximately 15% of the adducts. This reactivity is not observed in double-stranded DNA, rendering 1 a typical nonleaving group in reactions with this potential biological target. In enzymatic digests of calf thymus DNA treated with 2, three adducts were identified: [Pt(en)(ACRAMTU)(dGuo)](3+) (A1, approximately 80%), [Pt(en)(ACRAMTU)[d(GpA)]](2+) (A2, approximately 12%), and [Pt(en)(ACRAMTU)[d(TpA)]](2+) (A3, approximately 8%). A1 and P1 proved to be identical species. In the dinucleotide adducts A2 and A3, complex 2 covalently modifies adenine at GA and TA base steps, which are high-affinity intercalation sites of the acridine derivative 1. A2 and A3, which may be formed in the minor groove of DNA, are the first examples of monofunctional adenine adducts of divalent platinum formed in double-stranded DNA. The analysis of the adduct profile indicates that the sequence specificity of 1 plays an important role in the molecular recognition between DNA and the corresponding conjugate, 2. Possible biological consequences of the unusual adduct profile are discussed. PMID- 12904030 TI - Determinants of unnatural nucleobase stability and polymerase recognition. AB - Six new unnatural nucleobases have been synthesized and characterized in terms of stability and selectivity of self-pairing in duplex DNA and efficiency and fidelity of self-pairing during polymerase-mediated replication. Each nucleobase has a conserved ring structure but differs from the others in its specific pattern of substitution with oxygen and sulfur atoms. Heteroatom derivatization within the conserved scaffold is shown to have only moderate effects on unnatural self-pair synthesis by the polymerase; larger effects were observed on the thermal stability and polymerase-mediated extension of the self-pairs. The largest effects of heteroatom substitution were on the stability and synthesis of mispairs between the unnatural and natural bases. Certain heteroatom substitutions were found to have a general effect while others were found to have effects that were specific for a particular unnatural or natural base. The data are useful for designing stable and replicable third base pairs and for understanding the contributions of nucleobase shape, polarity, and polarizability to the stability and replication of DNA. PMID- 12904031 TI - Coordination-driven self-assembly of supramolecular cages: heteroatom-containing and complementary trigonal prisms. AB - The self-assembly of three nanoscopic prisms of approximate size 1 x 4 nm is reported. Tetrahedral carbon, silicon, and phosphorus were used as structure defining elements in these coordination-based cages. A carbon-based assembly completes a pair of nanoscopic complementary 3-D structures. The formation of the structures is supported by multinuclear NMR, ESI FT-ICR mass spectrometry, and elemental analysis data. PMID- 12904032 TI - Asymmetric synthesis of the highly potent anti-metastatic prostacyclin analogue cicaprost and its isomer isocicaprost. AB - An asymmetric synthesis of the anti-metastatic prostacyclin analogue cicaprost and a formal one of its isomer isocicaprost by a new route are described. A key step of these syntheses is the coupling of a chiral bicyclic C6-C14 ethynyl building block with a chiral C15-C21 omega-side chain amide building block with formation of the C14-C15 bond of the target molecules. A highly stereoselective reduction of the thereby obtained C6-C21 intermediate carrying a carbonyl group at C15 of the side chain was accomplished by the chiral oxazaborolidine method. The chiral phosphono acetate method was used for the highly stereoselective attachment of the alpha-side chain to the bicyclic C6-C21 intermediate carrying a carbonyl group at C6. Asymmetric syntheses of the bicyclic C6-C14 ethynyl building blocks were carried out starting from achiral bicyclic C6-C12 ketones by using the chiral lithium amide method. In the course of these syntheses, a new method for the introduction of an ethynyl group at the alpha-position of the carbonyl group of a ketone with formation of the corresponding homopropargylic alcohol was devised. Its key steps are an aldol reaction of the corresponding silyl enol ether with chloral and the elimination of a trichlorocarbinol derivative with formation of the ethynyl group. In addition, a new aldehyde to terminal alkyne transformation has been realized. Its key steps are the conversion of an aldehyde to the corresponding 1-alkenyl dimethylaminosulfoxonium salt and the elimination of the latter with a strong base. Two basically different routes have been followed for the synthesis of the enantiomerically pure C15-C21 omega-side chain amide building block. The first is based on the chiral oxazolidinone method and features a highly stereoselective alkylation of (4R)-N-acetyl-4-benzyloxazolidin-2-one, and the second encompasses a malonate synthesis of the racemic amide and its efficient preparative scale resolution by HPLC on a chiral stationary phase containing column. PMID- 12904033 TI - Efficient excitation energy transfer in long meso-meso linked Zn(II) porphyrin arrays bearing a 5,15-bisphenylethynylated Zn(II) porphyrin acceptor. AB - Electronically coupled porphyrin arrays are suitable for artificial light harvesting antenna in light of a large absorption cross-section and fast excitation energy transfer (EET). Along this line, an artificial energy transfer model system has been synthesized, comprising of an energy donating meso-meso linked Zn(II) porphyrin array and an energy accepting 5,15-bisphenylethynylated Zn(II) porphyrin linked via a 1,4-phenylene spacer. This includes an increasing number of porphyrins in the meso-meso linked Zn(II) porphyrin array, 1, 2, 3, 6, 12, and 24 (Z1A, Z2A, Z3A, Z6A, Z12A, and Z24A). The intramolecular singlet singlet EET processes have been examined by means of the steady-state and time resolved spectroscopic techniques. The steady-state fluorescence comes only from the acceptor moiety in Z1A-Z12A, indicating nearly the quantitative EET. In Z24A that has a molecular length of ca. 217 A, the fluorescence comes largely from the acceptor moiety but partly from the long donor array, indicating that the intramolecular EET is not quantitative. The transient absorption spectroscopy has provided the EET rates in real time scale: (2.5 ps)(-1) for Z1A, (3.3 ps)(-1) for Z2A, (5.5 ps)(-1) for Z3A, (21 ps)(-1) for Z6A, (63 ps)(-1) for Z12A, and (108 ps)(-1) for Z24A. These results have been well explained by a revised Forster equation (Sumi formula), which takes into account an exciton extending coherently over several porphyrin pigments in the donor array, whose length is not much shorter than the average donor-acceptor distance. Advantages of such strongly coupled porphyrin arrays in light harvesting and transmission are emphasized in terms of fast EET and a large absorption cross-section for incident light. PMID- 12904034 TI - Indium-mediated beta-allylation, beta-propargylation, and beta-allenylation onto alpha,beta-unsaturated ketones: reactions of in-situ-generated 3-tert butyldimethylsilyloxyalk-2-enylsulfonium salts with in-situ-generated organoindium reagents. AB - 3-tert-Butyldimethylsilyloxyalk-2-enylsulfonium salts, generated in situ from the reaction of alpha,beta-enones with dimethyl sulfide in the presence of TBSOTf, underwent a novel nucleophilic substitution with allylindiums to give silyl enol ethers of delta,epsilon-alkenyl ketones in good yields, which correspond to formal Michael addition products. In a similar manner, 1,4-propargylation of propargylindiums onto the sulfonium salts produced the corresponding silyl enol ethers of delta,epsilon-alkynyl ketones in good yields. Organoindium reagents derived from gamma-substituted propargyl bromide and indium afforded the corresponding silyl enol ethers of beta-allenyl ketones in good yields. The reaction proceeds via an addition-substitution mechanism involving the formation of allylic sulfonium salts. The presence of the intermediate sulfonium salt was confirmed by observation of the low-temperature (1)H NMR spectra. PMID- 12904035 TI - An ab initio study toward understanding the mechanistic photochemistry of acetamide. AB - The potential energy surfaces for CH(3)CONH(2) dissociation into CH(3) + CONH(2), CH(3)CO + NH(2), CH(3)CN + H(2)O, and CH(3)NH(2) + CO in the ground and lowest triplet states have been mapped with DFT, MP2, and CASSCF methods with the cc pVDZ and cc-pVTZ basis sets, while the S(1) potential energy surfaces for these reactions were determined by the CASSCF/cc-pVDZ optimizations followed by CASSCF/MRSDCI single-point calculations. The reaction pathways leading to different photoproducts are characterized on the basis of the computed potential energy surfaces and surface crossing points. A comparison of the reactivity among HCONH(2), CH(3)CONH(2), and CH(3)CONHCH(3) has been made, which provides some new insights into the mechanism of the ultraviolet photodissociation of small amides. PMID- 12904036 TI - Slow anion exchange, conformational equilibria, and fluorescent sensing in venus flytrap aminopyridinium-based anion hosts. AB - The synthesis, anion binding, and conformational properties of a series of 3 aminopyridinium-based, tripodal, tricationic hosts for anions are described. Slow anion and conformational exchange on the (1)H NMR time scale at low temperature, coupled with NMR titration, results in a high level of understanding of the anion binding properties of the compounds, particularly with respect to significant conformational change resulting from induced fit complexation. Peak selectivity for halides, particularly Cl(-), is observed. The approach has been extended to dipodal and tripodal podands based on 3-aminopyridinium "arms" containing photoactive anthracenyl moieties. The 1,3,5-tripodal host shows a remarkable selectivity for acetate over other anions, in contrast to the analogous unsubstituted tris(3-aminopyridinium) analogue, despite the fact that low temperature (1)H NMR experiments reveal a total of four acetate-binding conformations. Photodimerization of anthracene units results in the formation of potential fluorescent anion sensors. PMID- 12904037 TI - Self-assembly of ferrocene-functionalized perylene bisimide bridging ligands with PtII corner to electrochemically active molecular squares. AB - Ferrocenyl-substituted N,N'-di(4-pyridyl)perylene bisimide ligands have been synthesized by the coupling reaction of hydroxyphenoxy-perylene bisimides with ferrocenyl carboxylic acids. By means of metallosupramolecular self-assembly, hitherto unprecedented multiredox active dendritic molecular squares with 16 ferrocene groups positioned in the bridging ligands are prepared from the perylene bispyridyl imide ligands and [Pt(dppp)][(OTf)(2)] (dppp = 1,3 bis(diphenylphosphano)propane; OTf = trifluoromethanesulfonate) corner in high yield. The isolated metallosupramolecular squares were characterized by elemental analysis, (1)H, (31)P[(1)H] NMR, and UV/vis spectroscopy. The electrochemical properties of the ligands and squares are investigated by cyclic voltammetry as well as spectroelectrochemistry. The results obtained show that the redox behavior of ferrocene units is influenced by the square superstructure. Furthermore, redox titration of free ligand and corresponding molecular square with the one-electron oxidant thianthrenium pentachloroantimonate reveals that ferrocene groups in these structures may be oxidized completely by this oxidant, and highly charged species generated through oxidation of ferrocenyl groups in molecular square cause decomposition of the assembly due to pronounced Coulombic repulsion. PMID- 12904038 TI - Dehydrophenylnitrenes: quartet versus doublet states. AB - The geometries and energies of 4-, 3-, and 2-dehydrophenylnitrenes (3, 4, and 5) are investigated using complete active space self-consistent field (CASSCF), multiconfiguration quasi-degenerate second-order perturbation (MCQDPT), and internally contracted multiconfiguration-reference configuration interaction (MRCI) theories in conjunction with a correlation consistent triple-zeta basis set. 4-Dehydrophenylnitrene 3 has a quartet ground state ((4)A(2)). The adiabatic excitation energies to the (2)A(2), (2)B(2), (2)A(1), and (2)B(1) states are 5, 21, 34, and 62 kcal mol(-1), respectively. The (2)B(2) state has pronounced closed-shell carbene/iminyl radical character, while the lowest-energy (2)B(1) state is a combination of a planar allene and a 2-iminylpropa-1,3-diyl. The MCQDPT treatment overestimates the excitation energy to (2)B(2) significantly as compared to CASSCF and MRCI+Q. Among quartet states, (4)A(2)-3 is the most stable one, while those of 4 and 5 (both (4)A'') are 3 and 1 kcal mol(-1) higher in energy. 5 also has a quartet ground state and a (2)A' ' state 7 kcal mol(-1) higher in energy. On the other hand, the doublet-quartet energy splitting is -6 kcal mol(-1) for 4 in favor of the doublet state ((2)A''). Hence, (2)A''-4 is the most stable dehydrophenylnitrene, 3.5 kcal mol(-1) below (4)A(2) of 3. The geometry of (2)A''-4 shows the characteristic features of through-bond interaction between the in-plane molecular orbitals at N and at C3. The (2)A' state of 4 resembles the (2)A(1) state of 3 and lies 32 kcal mol(-1) above (4)A'' 4. The lowest-energy (2)A' state of 5, on the other hand, resembles the (2)B(2) state of 3 and lies 22 kcal mol(-1) above (4)A''-5. PMID- 12904039 TI - Ordered architectures of a soluble hexa-peri-hexabenzocoronene-pyrene dyad: thermotropic bulk properties and nanoscale phase segregation at surfaces. AB - An alkylated hexa-peri-hexabenzocoronene with a covalently tethered pyrene unit serves as a model to study self-assembling discotic pi-system dyads both in the bulk and at a surface. Wide-angle X-ray scattering, polarized light microscopy, and differential scanning calorimetry revealed bulk self-assembly into columnar structures. Relative to a control without a tethered pyrene, the new dyad exhibits a more ordered columnar phase at room temperature but with dramatically lowered isotropization temperature, facilitating homeotropic alignment. These two features are important for processing such materials into molecular electronic devices, e.g., photovoltaic diodes. Scanning tunneling microscopy at a solution solid interface revealed uniform nanoscale segregation of the large from the small pi-systems, leading to a well-defined two-dimensional crystalline monolayer, the likes of which may be employed in the future to study intramolecular electron transfer processes at surfaces, on the molecular scale. PMID- 12904040 TI - Structure-based design of a highly selective catalytic site-directed inhibitor of Ser/Thr protein phosphatase 2B (calcineurin). AB - Protein serine/threonine phosphatases (PP1, PP2A and PP2B) play important roles in intracellular signal transductions. The immunosuppressant drugs FK506 and cyclosporin A (CsA) bind to immunophilins, and these complexes selectively inhibit PP2B (calcineurin), leading to the suppression of T-cell proliferation. Both FK506 and CsA must, however, form complexes with immunophilins to exert their inhibitory action on PP2B. Thus, it is of interest to find a direct and selective inhibitor of PP2B that does not involve the immunophilins as a biological tool for studies of PP2B and also as a candidate therapeutic agent. We selected the simple natural product cantharidin, a known PP2A-selective inhibitor, as a lead compound for this project. Primary SAR indicated that norcantharidin (7-oxabicyclo[2.2.1]heptane-2,3-dicarboxylic anhydride) inhibits not only PP1 and PP2A but also PP2B, and a binding model of norcantharidin carboxylate to the PP2B catalytic site was computationally constructed. Based on this binding model, we designed and synthesized several cantharidin derivatives. Among these compounds, 1,5-dibenzoyloxymethyl-substituted norcantharidin was found to inhibit PP2B without inhibiting PP1 or PP2A. To our knowledge, this is the first highly selective catalytic site-directed inhibitor of PP2B. PMID- 12904041 TI - Mechanism of a strongly anisotropic MoIII-CN-MnII spin-spin coupling in molecular magnets based on the [Mo(CN)(7)](4-) heptacyanometalate: a new strategy for single-molecule magnets with high blocking temperatures. AB - Unusual spin coupling between Mo(III) and Mn(II) cyano-bridged ions in bimetallic molecular magnets based on the [Mo(III)(CN)(7)](4-) heptacyanometalate is analyzed in terms of the superexchange theory. Due to the orbital degeneracy and strong spin-orbit coupling on Mo(III), the ground state of the pentagonal bipyramidal [Mo(III)(CN)(7)](4-) complex corresponds to an anisotropic Kramers doublet. Using a specially adapted kinetic exchange model we have shown that the Mo(III)-CN-Mn(II) superexchange interaction is extremely anisotropic: it is described by an Ising-like spin Hamiltonian JS(z)(Mo) S(z)(Mn) for the apical pairs and by the J(z)S(z)(Mo) S(z)(Mn) + J(xy)(Sx(Mo) Sx(Mn) + Sy(Mo) Sy(Mn)) spin Hamiltonian for the equatorial pairs (in the latter case J(z) and J(xy) can have opposite signs). This anisotropy resulted from an interplay of several Ising like (Sz(Mo) Sz(Mn)) and isotropic (S(Mo)S(Mn)) ferro- and antiferromagnetic contributions originating from metal-to-metal electron transfers through the pi and sigma orbitals of the cyano bridges. The Mo(III)-CN-Mn(II) exchange anisotropy is distinct from the anisotropy of the g-tensor of [Mo(III)(CN)(7)](4 ); moreover, there is no correlation between the exchange anisotropy and g-tensor anisotropy. We indicate that highly anisotropic spin-spin couplings (such as the Ising-like JS(z)(Mo) S(z)(Mn)) combined with large exchange parameters represent a very important source of the global magnetic anisotropy of polyatomic molecular magnetic clusters. Since the total spin of such clusters is no longer a good quantum number, the spin spectrum pattern can differ considerably from the conventional scheme described by the zero-field splitting of the isotropic spin of the ground state. As a result, the spin reorientation barrier of the magnetic cluster may be considerably larger. This finding opens a new way in the strategy of designing single-molecule magnets (SMM) with unusually high blocking temperatures. The use of orbitally degenerate complexes with a strong spin-orbit coupling (such as [Mo(III)(CN)(7)](4-) or its 5d analogues) as building blocks is therefore very promising for these purposes. PMID- 12904042 TI - Individual single-walled nanotubes and hydrogels made by oxidative exfoliation of carbon nanotube ropes. AB - Single-walled carbon nanotubes were oxidized by a technique previously developed for the oxidation of graphite to graphite oxide (GO). This process involves treatment with concentrated H(2)SO(4) containing (NH(4))(2)S(2)O(8) and P(2)O(5), followed by H(2)SO(4) and KMnO(4). Oxidation results in complete exfoliation of nanotube ropes to yield individual oxidized tubes that are 40-500 nm long. The C:O:H atomic ratio of vacuum-dried oxidized nanotubes is approximately 2.7:1.0:1.2. XPS and IR spectra show evidence for surface O-H, C=O, and COOH groups. The oxidized nanotubes slowly form viscous hydrogels at unusually low concentration (>or=0.3 wt %), and this behavior is attributed to the formation of a hydrogen-bonded nanotube network. The oxidized tubes bind readily to amine coated surfaces, on which they adsorb as smooth and dense monolayer films. Thin films of the oxidized nanotubes show ohmic current-voltage behavior, with resistivities in the range of 0.2-0.5 Omega-cm. PMID- 12904043 TI - Controlled formation of highly organized mesoporous titania thin films: from mesostructured hybrids to mesoporous nanoanatase TiO2. AB - In this paper, we report the complete synthesis and characterization procedures to generate highly organized and oriented mesoporous titania thin films, using poly(ethylene oxide) (PEO)-based templates. Controlled conditions in the deposition, postsynthesis, and thermal treatment steps allow one to tailor the final mesostructure (2D hexagonal, p6m, or 3D cubic, Im3m). Various techniques were used to determine the time evolution of the mesostructure. Spectroscopic techniques (UV/vis, (17)O NMR) and EXAFS/XANES have been used to follow the chemical changes in the Ti(IV) environment. Crossing these techniques spanning all ranges permits a complete description of the chemistry all the way from solution to the mesostructured metal oxide. A critical discussion on all important chemical and processing parameters is provided; the understanding of these features is essential for a rational design and the reproducible construction of mesoporous materials. PMID- 12904044 TI - Photoinduced axial ligation and deligation dynamics of nonplanar nickel dodecaarylporphyrins. AB - The ground- and excited-state metal-ligand dynamics of nonplanar nickel(II) 2,3,5,7,8,10,12,13,15,17,18,20-dodecaphenylporphyrin (NiDPP) and two fluorinated analogues (NiF(20)DPP and NiF(28)DPP) have been investigated using static and time-resolved absorption spectroscopy in toluene and in ligating media that differ in basicity, aromaticity, and steric encumbrance. Because of the electronic and steric consequences of nonplanarity, NiDPP does not bind axial ligands in the ground state, but metal coordination does occur after photoexcitation with multistep dynamics that depend on the properties of the ligand. Following the structural relaxations that occur in all nickel porphyrins within approximately 10 ps, ligand binding to photoexcited NiDPP is progressively longer in pyridine, piperidine, and 3,5-lutidine (25-100 ps) but does not occur at all in 2,6-lutidine in which the ligating nitrogen is sterically encumbered. The transient intermediate that is formed, which nominally could be either a five or six-coordinate species, also has a ligand-dependent lifetime (200-550 ps). Decay of this intermediate occurs partially via ligand release to re-form the uncoordinated species, in competition with binding of the second axial ligand and/or conformational/electronic relaxations (of a six-coordinate intermediate) to give the ground state of the bis-ligated photoproduct. The finding that the photoproduct channel principally depends on ligand characteristics along with the time-evolving spectra suggests that the transient intermediate may involve a five coordinate species. In contrast to NiDPP, the fluorinated analogues NiF(20)DPP and NiF(28)DPP do coordinate axial ligands in the ground state but eject them after photoexcitation. Collectively, these results demonstrate the sensitivity with which the electronic and structural characteristics of the macrocycle, substituents, and solvent (ligands) can govern the photophysical and photochemical properties of nonplanar porphyrins and open new avenues for exploring photoinduced ligand association and dissociation behavior. PMID- 12904045 TI - A novel and general synthetic pathway to strychnos indole alkaloids: total syntheses of (-)-tubifoline, (-)-dehydrotubifoline, and (-)-strychnine using palladium-catalyzed asymmetric allylic substitution. AB - A method of palladium-catalyzed asymmetric allylic substitution for synthesizing 2-substituted cyclohexenylamine derivatives was established. Treatment of a 2 silyloxymethylcyclohexenol derivative with ortho-bromo-N-tosylaniline in the presence of Pd(2)dba(3).CHCl(3) and (S)-BINAPO in THF afforded a cyclohexenylamine derivative with 84% ee in 80% yield. The Heck reaction was carried out to produce an indolenine derivative in good yield. Using this method, we synthesized indolenine derivative 7, which was recrystallized from EtOH to give an optically pure compound. From this compound, tetracyclic ketone 13, which should be a useful intermediate for the synthesis of indole alkaloids, could be synthesized. The total syntheses of (-)-dehydrotubifoline, (-)-tubifoline, and ( )-strychnine were achieved from 13. All ring constructions for the syntheses of these natural products were achieved using a palladium catalyst. PMID- 12904046 TI - A mechanistic investigation of the carbon-carbon bond cleavage of aryl and alkyl cyanides using a cationic RhIII silyl complex. AB - Cationic Rh(III) complex [Cp(PMe(3))Rh(SiPh(3))(CH(2)Cl(2))]BAr(4)' (1) activates the carbon-carbon bond of aryl and alkyl cyanides (R-CN, where R = Ph, (4 (CF(3))C(6)H(4)), (4-(OMe)C(6)H(4)), Me, (i)Pr, (t)Bu) to produce complexes of the general formula [Cp*(PMe(3))Rh(R)(CNSiPh(3))]BAr(4)'. With the exception of the (t)BuCN case, every reaction proceeds at room temperature (t(1/2) < 1 h for aryl cyanides, t(1/2) < 14 h for alkyl cyanides). A general mechanism is presented on the basis of (1) an X-ray crystal structure determination of an intermediate isolated from the reaction involving 4-methoxybenzonitrile and (2) kinetic studies performed on the C-C bond cleavage of para-substituted aryl cyanides. Initial formation of an eta(1)-nitrile species is observed, followed by conversion to an eta(2)-iminoacyl intermediate, which was observed to undergo migration of R (aryl or alkyl) to rhodium to form the product [Cp*(PMe(3))Rh(R)(CNSiPh(3))]BAr(4)'. PMID- 12904047 TI - Photodissociation dynamics of fluorobenzene. AB - Photodissociation of both fluorobenzene and d(5)-fluorobenzene at 193 nm under collision-free conditions has been studied in separate experiments using multimass ion imaging techniques. HF and DF eliminations were found to be the major dissociation channels. Small amounts of photofragments, C(6)H(4)F and C(6)D(4)F, corresponding to H and D atom eliminations were also observed. Dissociation rate and fragment translational energy distribution suggest that HF (DF) and H (D) atom elimination reactions occur in the ground electronic state. The potential energy surface obtained from ab initio calculations indicates that the four-center reaction in the ground electronic state is the major dissociation mechanism for the HF and DF eliminations. A comparison with photodissociation of benzene has been made. PMID- 12904048 TI - Ab initio calculation of the vibrational and electronic spectra of trans- and cis azobenzene. AB - We report accurate geometries and harmonic force fields for trans- and cis azobenzene determined by second-order Moller-Plesset perturbation theory. For the trans isomer, the planar structure with C(2h) symmetry, found in a recent gas electron diffraction experiment, is verified. The calculated vibrational spectra are compared with experimental data and density functional calculations. Important vibrational frequencies are localized and discussed. For both isomers, we report UV spectra calculated using the second-order approximate coupled cluster singles-and-doubles model CC2 with accurate basis sets. Vertical excitation energies and oscillator strengths have been determined for the lowest singlet n(pi)* and (pi)(pi)* transitions. The results are compared with the available experimental data and second-order polarization propagator (SOPPA) and density functional (DFT) calculations. For both isomers, the CC2 results for the excitation energies into the S(1) and S(2) states agree within 0.1 eV with experimental gas-phase measurements. PMID- 12904049 TI - Effects of surface coordination chemistry on the magnetic properties of MnFe(2)O(4) spinel ferrite nanoparticles. AB - To understand the influence of surface interactions upon the magnetic properties of magnetic nanoparticles, the surface of manganese ferrite, MnFe(2)O(4), nanoparticles have been systematically modified with a series of para-substituted benzoic acid ligands (HOOC-C(6)H(4)-R; R = H, CH(3), Cl, NO(2), OH) and substituted benzene ligands (Y-C(6)H(5), Y = COOH, SH, NH(2), OH, SO(3)H). The coercivity of magnetic nanoparticles decreases up to almost 50% upon the coordination of the ligands on the nanoparticle surface, whereas the saturation magnetization has increased. The percentage coercivity decrease of the modified nanoparticles with respect to the native nanoparticles strongly correlates with the crystal field splitting energy (CFSE) Delta evoked by the coordination ligands. The ligand inducing largest CFSE results in the strongest effect on the coercivity of magnetic nanoparticles. The change in magnetic properties of nanoparticles also correlates with the specific coordinating functional group bound onto the nanoparticle surface. The correlations suggest the decrease in spin-orbital couplings and surface anisotropy of magnetic nanoparticles due to the surface coordination. Such surface effects clearly show the dependence on the size of nanoparticles. PMID- 12904051 TI - Solvent reorganization in electron and ion transfer reactions near a smooth electrified surface: a molecular dynamics study. AB - Molecular dynamics simulations of electron and ion transfer reactions near a smooth surface are presented, analyzing the effect of the geometrical constraint of the surface and the interfacial electric field on the relevant solvation properties of both a monovalent negative ion and a neutral atom. The simulations show that, from the solvation point of view, ion adsorption is an uphill process due to the need to shed off the ion's solvation shell and displace water from the surface. Atom adsorption, on the other hand, has only a small barrier, related to the molecularity of the solvent. Both the electrostatic interaction of the ion with the solvent and the ion's solvent reorganization energy (the relevant parameter in the Marcus electron transfer theory) decrease as the surface is approached, whereas these parameters are not sensitive to the distance from the surface for the atom. This is a consequence of the importance of long-range electrostatic interactions for ion solvation and the importance of short-range interactions for atom solvation. The electric field either attracts or repels an ion to or from the surface, but the field has no influence on the solvent reorganization energy. By including the quantum-mechanical electron transfer between the metal surface and the ion/atom in solution in the MD simulation by using a model Hamiltonian, we calculated two-dimensional free energy surfaces for ion adsorption allowing for partial charge transfer, based on a fully molecular picture of ion solvation near the surface. PMID- 12904050 TI - Multicomponent submicron features of biomolecules created by voltage controlled deposition from a nanopipet. AB - We have used a nanopipet as a nanopen to locally and controllably deposit complex biomolecules, including antibodies and DNA, onto a surface to create multicomponent and functional submicron features. Key advantages of this method are that the biomolecules are always in solution and the applied voltage provides fine control of the delivery down to the single molecule level. Two consecutive cycles of deposition, to produce spatially varying features with different biological properties, were demonstrated with fluorescently labeled antibodies or biotin. This approach combines "top-down" fabrication, using the nanopen for local application, and "bottom-up" fabrication, using molecular recognition for self-assembly at defined positions, and opens up new possibilities in nanotechnology. PMID- 12904052 TI - On the electronic origins of structural isomerism in the iron-sulfur cubane, [(C(5)H(5))(4)Fe(4)S(4)](2+). AB - Density functional theory provides new insights into the structural isomerism observed in the cyclopentadienyl-capped iron-sulfur cluster, [(C(5)H(5))(4)Fe(4)S(4)](2+). Two distinct, closely spaced minima have been located, a triplet with D(2) symmetry and a C(2)-symmetric singlet, both of which correspond closely to the structure of one of the known crystal forms of the cation. Thus, the structural diversity in these species reflects genuine molecular bistability rather than simple solid-state packing effects. In contrast, no stable D(2)(d)()-symmetric minimum has been located, suggesting that the reported D(2)(d)() symmetry of the cation in [(C(5)H(5))(4)Fe(4)S(4)][PF(6)](2) may be a crystallographic artifact. In the ruthenium analogue, the more diffuse 4d orbitals stabilize the C(2)-symmetric singlet, which is unambiguously the ground state, but the D(2)-symmetric potential energy surface provides a viable low-energy pathway for the dynamic exchange of the Ru-Ru bonds. PMID- 12904053 TI - A new angle on heat capacity changes in hydrophobic solvation. AB - The differential solubility of polar and apolar groups in water is important for the self-assembly of globular proteins, lipid membranes, nucleic acids, and other specific biological structures through hydrophobic and hydrophilic effects. The increase in water's heat capacity upon hydration of apolar compounds is one signature of the hydrophobic effect and differentiates it from the hydration of polar compounds, which cause a decrease in heat capacity. Water structuring around apolar and polar groups is an important factor in their differential solubility and heat capacity effects. Here, it is shown that joint radial/angular distribution functions of water obtained from simulations reveal quite different hydration structures around polar and apolar groups: polar and apolar groups have a deficit or excess, respectively, of "low angle hydrogen bonds". Low angle hydrogen bonds have a larger energy fluctuation than high angle bonds, and analysis of these differences provides a physical reason for the opposite changes in heat capacity and new insight into water structure around solutes and the hydrophobic effect. PMID- 12904054 TI - Theoretical examination of Mg(2+)-mediated hydrolysis of a phosphodiester linkage as proposed for the hammerhead ribozyme. AB - The hammerhead ribozyme is an RNA molecule capable of self-cleavage at a unique site within its sequence. Hydrolysis of this phosphodiester linkage has been proposed to occur via an in-line attack geometry for nucleophilic displacement by the 2'-hydroxyl on the adjoining phosphorus to generate a 2',3'-cyclic phosphate ester with elimination of the 5'-hydroxyl group, requiring a divalent metal ion under physiological conditions. The proposed S(N)2(P) reaction mechanism was investigated using density functional theory calculations incorporating the hybrid functional B3LYP to study this metal ion-dependent reaction with a tetraaquo magnesium (II)-bound hydroxide ion. For the Mg(2+)-catalyzed reaction, the gas-phase geometry optimized calculations predict two transition states with a kinetically insignificant, yet clearly defined, pentacoordinate intermediate. The first transition state located for the reaction is characterized by internal nucleophilic attack coupled to proton transfer. The second transition state, the rate-determining step, involves breaking of the exocyclic P-O bond where a metal ligated water molecule assists in the departure of the leaving group. These calculations demonstrate that the reaction mechanism incorporating a single metal ion, serving as a Lewis acid, functions as a general base and can afford the necessary stabilization to the leaving group by orienting a water molecule for catalysis. PMID- 12904056 TI - Theoretical and experimental investigations on the chemical reactions of positively charged phenyl radicals with cytosine and 1-methylcytosine. AB - The chemical behavior of positively charged phenyl radicals toward cytosine, 1 methylcytosine, and some pyrimidine analogues in the gas phase was investigated both theoretically by performing molecular orbital calculations and experimentally by using FT/ICR mass spectrometry. The phenyl radicals react with cytosine and 1-methylcytosine predominately by hydrogen abstraction and addition. For cytosine, the preferred site for hydrogen abstraction appears to be the amino group, and addition occurs preferentially at the N3 and N1 positions of the keto and enol tautomeric forms, respectively. For 1-methylcytosine, the methyl group is the thermodynamically favored site for hydrogen abstraction and N3 for addition. Possible structures and formation mechanisms are suggested for two unknown product ions formed upon the reaction of cytosine with the 3-dehydro-N phenylpyridinium radical cation. PMID- 12904055 TI - Structure of gramicidin a in a lipid bilayer environment determined using molecular dynamics simulations and solid-state NMR data. AB - Two different high-resolution structures recently have been proposed for the membrane-spanning gramicidin A channel: one based on solid-state NMR experiments in oriented phospholipid bilayers (Ketchem, R. R.; Roux, B.; Cross, T. A. Structure 1997, 5, 1655-1669; Protein Data Bank, PDB:1MAG); and one based on two dimensional NMR in detergent micelles (Townsley, L. E.; Tucker, W. A.; Sham, S.; Hinton, J. F. Biochemistry 2001, 40, 11676-11686; PDB:1JNO). Despite overall agreement, the two structures differ in peptide backbone pitch and the orientation of several side chains; in particular that of the Trp at position 9. Given the importance of the peptide backbone and Trp side chains for ion permeation, we undertook an investigation of the two structures using molecular dynamics simulation with an explicit lipid bilayer membrane, similar to the system used for the solid-state NMR experiments. Based on 0.1 micros of simulation, both backbone structures converge to a structure with 6.25 residues per turn, in agreement with X-ray scattering, and broad agreement with SS backbone NMR observables. The side chain of Trp 9 is mobile, more so than Trp 11, 13, and 15, and undergoes spontaneous transitions between the orientations in 1JNO and 1MAG. Based on empirical fitting to the NMR results, and umbrella sampling calculations, we conclude that Trp 9 spends 80% of the time in the 1JNO orientation and 20% in the 1MAG orientation. These results underscore the utility of molecular dynamics simulations in the analysis and interpretation of structural information from solid-state NMR. PMID- 12904058 TI - Autoradiographic visualization of corticotropin releasing hormone type 1 receptors with a nonpeptide ligand: synthesis of [(76)Br]MJL-1-109-2. AB - A high-affinity, nonpeptide radioligand for the CRHR1 was synthesized and showed distribution in rat brain consistent with CRHR1 using in vitro autoradiography. This is the first nonpeptide radiotracer combining high affinity and appropriate lipophilicity that penetrates the blood-brain barrier and hence has the potential to be used for PET imaging studies. In vivo visualization of changes in the CRH1 receptor or its occupancy would further the understanding of the pathophysiology of stress related diseases. PMID- 12904059 TI - Antimalarial drug quinacrine binds to C-terminal helix of cellular prion protein. AB - Using NMR spectroscopy we show that the cellular prion protein constitutes a target for binding of various acridine and phenothiazine derivatives. We unambiguously map the quinacrine binding site of recombinant human prion protein to residues Tyr225, Tyr226, and Gln227 of helix alpha3, which is located near the "protein X" epitope. The millimolar dissociation constant of the complex suggests that in vivo inhibition of prion propagation occurs after 10000-fold concentration of quinacrine within endolysosomes. PMID- 12904060 TI - Selective formation of homo- and heterobivalent peptidomimetics. AB - Methodology is presented for assembling fluorescently labeled bivalent molecules from monovalent constituents, without side chain protection or coupling agents. To illustrate the procedure, a series of bivalent peptidomimetics directed toward the Trk receptors were prepared and screened via fluorescent activated cell sorting scan assays. PMID- 12904061 TI - Structure-activity relationships in carboxamide derivatives based on the targeted delivery of radionuclides and boron atoms by means of peripheral benzodiazepine receptor ligands. AB - The structure-activity relationship studies on 2-quinolinecarboxamide peripheral benzodiazepine receptor (PBR) ligands have been refined with the aim of using these ligands as carriers of radionuclides and boron atoms. Some new ligands show enhanced affinity and steroidogenic activity with respect to reference compound 1 and are interesting candidates for radiolabeling and PET studies. Moreover, carborane derivative 3q, representing the first example of PBR ligand bearing a carborane cage, can be useful to explore an alternative mechanism in BNCT. PMID- 12904062 TI - Prediction of aqueous solubility of a diverse set of compounds using quantitative structure-property relationships. AB - "Fail early and fail fast" is the current paradigm that the pharmaceutical industry has adopted widely. Removing non-drug-like compounds from the drug discovery lifecycle in the early stages can lead to tremendous savings of resources. Thus, fast screening methods are needed to profile the large collection of synthesized and virtual libraries involved in the early stage. Solubility is one of the filters that are applied extensively to ensure that the compounds are reasonably soluble so that synthesis of the compounds and assay studies of pharmacokinetics and toxicity are feasible. To address this need, we have developed a fast quantitative structure-property relationship (QSPR) model for the prediction of aqueous solubility (at 298 K, unbuffered solution) from the molecular structures. Multiple linear regressions and genetic algorithms were used to develop the models. The model was based on a set of diverse compounds including small organic molecules and drug and drug-like species. The predicted solubility for the training and test sets agrees well with the experimental values. The coefficient of determination is R(2) = 0.84 for the training set of 775 compounds and the RMS error = 0.87. This model was validated on four sets of compounds. The RMS error for the 1665 compounds from the four validation data sets (including compounds from the Physician's Desk References and Comprehensive Medicinal Chemistry databases) is 1 log unit and the unsigned error is 0.77. This model does not require 3-D structure generation which is rather time-consuming. Using 2-D structure as input, this model is able to compute solubility for 90 000 700 000 compounds/h on a SGI Origin 2000 workstation. This kind of fast calculation allows the model to be used in data mining and screening of large synthesized or virtual libraries. PMID- 12904063 TI - Design, synthesis, and evaluation of substituted phenylpropanoic acid derivatives as human peroxisome proliferator activated receptor activators. Discovery of potent and human peroxisome proliferator activated receptor alpha subtype selective activators. AB - Substituted phenylpropanoic acid derivatives were prepared as part of a search for subtype-selective human peroxisome proliferator activated receptor alpha (PPARalpha) activators. Structure-activity relationship studies indicated that the nature and the stereochemistry of the substituent at the alpha-position of the head part containing the carboxyl group, the distance between the carboxyl group and the central benzene ring, the linking group between the central benzene ring and the distal benzene ring, and the substituent at the distal hydrophobic tail part of the molecule all play key roles in determining the potency and selectivity of PPAR subtype transactivation. This study has led to the identification of potent and human PPARalpha selective optically active alpha alkylphenylpropanoic acid derivatives, which will be useful not only as pharmacological tools to investigate the physiology and pathophysiology of PPARalpha but also as candidate drugs for the treatment of altered metabolic homeostasis, such as dyslipidemia, obesity, and diabetes. PMID- 12904064 TI - Functionalized glycomers as growth inhibitors and inducers of apoptosis in human glioblastoma cells. AB - The effects of functionalized aryl beta-D-glycopyranosides (glycomers) on the proliferation, survival, and apoptosis of human glioblastoma cells in culture were evaluated as a way to control tumor progression. The results showed that inhibition of growth and/or induction of apoptosis can be achieved by these molecules in human glioblastoma cells. Inhibition of DNA synthesis precedes induction of apoptosis and growth inhibition. The substituents at C-1, C-2, C-3,C 4, and C-6 on the pyranosidic scaffold are important to modulate the action and the efficacy of these molecules. Human fibroblasts and brain-derived endothelial cells were less sensitive to glycomers than tumor cells. Thus, functionalized aryl beta-D-glycopyranosides represent a new class of molecules potentially able to control the progression of brain tumors. PMID- 12904065 TI - Efficacious and orally bioavailable thrombin inhibitors based on a 2,5 thienylamidine at the P1 position: discovery of N-carboxymethyl-d-diphenylalanyl l-prolyl[(5-amidino-2-thienyl)methyl]amide. AB - Thrombin, a crucial enzyme in the blood coagulation, has been a target for antithrombotic therapy. Orally active thrombin inhibitors would provide effective and safe prophylaxis for venous and arterial thrombosis. We conducted optimization of a highly efficacious benzamidine-based thrombin inhibitor LB30812 (3, K(i) = 3 pM) to improve oral bioavailability. Of a variety of arylamidines investigated at the P1 position, 2,5-thienylamidine effectively replaced the benzamidine without compromising the thrombin inhibitory potency and oral absorption. The sulfamide and sulfonamide derivatization at the N-terminal position in general afforded highly potent thrombin inhibitors but with moderate oral absorption, while the well-absorbable N-carbamate derivatives exhibited limited metabolic stability in S9 fractions. The present work culminated in the discovery of the N-carboxymethyl- and 2,5-thienylamidine-containing compound 22 that exhibits the most favorable profiles of anticoagulant and antithrombotic activities as well as oral bioavilability (K(i) = 15 pM; F = 43%, 42%, and 15% in rats, dogs, and monkeys, respectively). This compound on a gravimetric basis was shown to be more effective than a low molecular weight heparin, enoxaparin, in the venous thrombosis models of rat and rabbit. Compound 22 (LB30870) was therefore selected for further preclinical and clinical development. PMID- 12904066 TI - Synthesis of novel macrocyclic docetaxel analogues. Influence of their macrocyclic ring size on tubulin activity. AB - This work describes the synthesis of a series of novel macrocyclic taxoids 3 and 3(H) designed to mimic the docetaxel solid-state ("nonpolar") conformation. These compounds, bearing 18-, 20-, 21-, and 22-membered rings connecting the C-2 OH and C-3' NH moieties, were constructed by ring-closing olefin metathesis of the taxoid-omega,omega'-dienes 4. Biological evaluation of these new taxoids showed that activity is dependent on the ring size, and only the 22-membered ring taxoid 3d exhibits significant tubulin binding. Synthesis of the open-chain analogues 7 and 7(H) and comparison of their biological activities with macrocyclic taxoids show that the carbon tether between C-2 OH and C-3' NH does not hamper tubulin binding. Computational studies of the conformational behavior of the macrocyclic taxoids 3 indicate that the 18-, 20-, and 21-membered-ring 3a-c adopt mainly conformations that are not recognized by tubulin. The most active taxoid 3d appears to adopt a conformation that is between the "nonpolar" and T-shaped forms. PMID- 12904067 TI - Modeling of human cytochrome p450-mediated drug metabolism using unsupervised machine learning approach. AB - We developed a computational algorithm for evaluating the possibility of cytochrome P450-mediated metabolic transformations that xenobiotics molecules undergo in the human body. First, we compiled a database of known human cytochrome P-450 substrates, products, and nonsubstrates for 38 enzyme-specific groups (total of 2200 compounds). Second, we determined the cytochrome-mediated metabolic reactions most typical for each group and examined the substrates and products of these reactions. To assess the probability of P450 transformations of novel compounds, we built a nonlinear quantitative structure-metabolism relationships (QSMR) model based on Kohonen self-organizing maps (SOM). This neural network QSMR model incorporated a predefined set of physicochemical descriptors encoding the key molecular properties that define the metabolic fate of individual molecules. Isozyme-specific groups of substrate molecules were visualized, thus facilitating prediction of tissue-specific metabolism. The developed algorithm can be used in early stages of drug discovery as an efficient tool for the assessment of human metabolism and toxicity of novel compounds in designing discovery libraries and in lead optimization. PMID- 12904068 TI - Structure-based design and synthesis of novel potent Na+,K+ -ATPase inhibitors derived from a 5alpha,14alpha-androstane scaffold as positive inotropic compounds. AB - The design, synthesis, and biological properties of novel inhibitors of the Na(+),K(+)-ATPase as potential positive inotropic compounds are reported. Following our model of superposition between cassaine and digitoxigenin, digitalis-like activity has been elicited from a non-digitalis steroidal structure by suitable modifications of the 5alpha,14alpha-androstane skeleton. The strong hydrophobic interaction of the digitalis or cassaine polycyclic cores can be effectively obtained with the androstane skeleton taken in a reversed orientation. Thus, oxidation of C-6 and introduction in the C-3 position of the potent pharmacophoric group recently introduced by us, in the 17 position of the digitalis skeleton, namely, O-(omega-aminoalkyl)oxime, led to a series of substituted androstanes able to inhibit the Na(+),K(+)-ATPase, most of them with an IC(50) in the low micromolar level, and to induce a positive inotropic effect in guinea pig. Within this series, androstane-3,6,17-trione (E,Z)-3-(2 aminoethyl)oxime (22b, PST 2744) induced a strong positive inotropic effect while being less arrhythmogenic than digoxin, when the two compounds were compared at equiinotropic doses. PMID- 12904069 TI - Discovery of (3S)-amino-(4R)-ethylpiperidinyl quinolones as potent antibacterial agents with a broad spectrum of activity and activity against resistant pathogens. AB - Novel quinolone antibacterial agents bearing (3S)-amino-(4R)-ethylpiperidines were designed by using low energy conformation analysis and synthesized by applying a conventional coupling reaction of the quinolone nuclei with new piperidine side chains. These compounds were tested in MIC assays and found to be highly potent against Gram-positive and Gram-negative organisms. In particular, the new compounds exhibited high activity against the resistant pathogens Staphylococcus aureus (MRCR) and Streptococcus pneumoniae (PR). Importantly, when the (3S)-amino-(4R)-ethylpiperidinyl quinolones were compared with marketed quinolones sharing the same quinolone nuclei but different side chains at the C-7 position, the new quinolones showed superior activity against Gram-positive organisms, including resistant pathogens. PMID- 12904070 TI - Synthesis and biological evaluation of 3-alkoxy analogues of flavone-8-acetic acid. AB - New analogues of flavone-8-acetic acid were synthesized, bearing an alkoxy group in position 3 and different substituents on the benzene ring in position 2 of the flavone nucleus. The compounds were tested for direct cytotoxicity against four human tumor cell lines and for indirect antitumor effects by measuring their ability to enhance lytic properties of murine macrophages and human monocytes. Though direct toxicity was very low, the compounds were able to induce significant indirect toxicity. Notably, most of them (4c, 4d, 4e, 4f, 4h, 4i, 4m,4n, and 4o) showed important activity on human monocytes and could be regarded as the first flavone derivatives endowed with such activity. Particularly interesting seem to be compounds 4m and 4n, which showed IC(50) values up to 7 times higher than DMXAA, which has now completed phase I clinical trials. PMID- 12904071 TI - Highly potent 1,4-benzothiazine derivatives as K(ATP)-channel openers. AB - A series of 1,4-benzothiazines, suitably functionalized at the N-4 and C-6 positions, arising from the replacement of a benzopyran-based structure of cromakalim with a 1,4-benzothiazine nucleus, has been synthesized as potassium channel openers (KCOs). Most of the tested compounds show high vasorelaxant potency that is considerably higher than that of the reference levcromakalim (LCRK). In the presence of the well-established selective K(ATP) blocker, glibenclamide, the vasorelaxing effects were antagonized in a competitive fashion, indicating the involvement of the K(ATP) channel in their pharmacological effect. Some aspects of the structure-activity relationship associated with the N-4 and C-6 substituents are discussed. The highest level of activity was achieved with a cyclopentenone ring at the N-4 position coupled with an electron-withdrawing group such as nitro, trifluoromethyl, or cyano at the C-6 position. Compounds 4c, 5c, and 6c displayed a vasorelaxant potency at least 10 000 times greater than that of LCRK, thus becoming the most potent KCOs identified to date. PMID- 12904072 TI - Design and synthesis of novel pyridoxine 5'-phosphonates as potential antiischemic agents. AB - On the basis of previous reports that the natural cofactor pyridoxal 5'-phosphate 1 appears to display cardioprotective properties, a series of novel mimetics of this cofactor were envisioned. As pyridoxal 5'-phosphate is a natural compound and is subject to biological degradation and elimination pathways, the objective was to generate active phosphonates that are potentially less light sensitive and more stable in vivo than the parent vitamer. Several phosphonates were designed and synthesized, and in particular, compounds 10 and 14 displayed similar biological traits to natural phosphate 1 in the rat model of regional myocardial ischemia and reperfusion. A reduction in infarct size was observed in animals treated with these compounds. In an effort to identify other relevant cardioprotective models in order to potentially define structure-activity relationships, these three compounds were tested in the rat working heart model. Compounds 1, 10, and 14 were compared to dichloroacetic acid (DCA) as positive control in this model. As with DCA, compounds 1, 10, and 14 were found to induce a shift from fatty acid oxidation toward glucose oxidation. PMID- 12904073 TI - Stereoselective synthesis and structure-activity relationship of novel ceramide trafficking inhibitors. (1R,3R)-N-(3-hydroxy-1-hydroxymethyl-3 phenylpropyl)dodecanamide and its analogues. AB - New ceramide trafficking inhibitors, (1R,3R)-N-(3-hydroxy-1-hydroxymethyl-3 phenylpropyl)dodecanamide (HPA-12) and a series of its analogues, were synthesized in diastereomerically and enantiomerically pure forms, and the structure-activity relationship was investigated. These analogues were stereoselectively synthesized via catalytic enantioselective Mannich-type reactions using a Cu(II)-chiral diamine 4 complex. Analysis of HPA-12 analogues having various lengths of the amide side chain showed that the optimal chain length for the inhibition of sphingomyelin biosynthesis is 13 with an IC(50) of approximately 50 nM. Masking of the hydroxy group at the 2'- or 3-position of HPA 12 was carried out by methylation, and it was revealed that these hydroxy groups were essential for the activity. Installation of another hydroxy group onto HPA 12 at the same position as that in the natural ceramide was also conducted, but no enhancement of the activity was observed. PMID- 12904074 TI - Enantioselective syntheses and cytotoxicity of N,O-nucleosides. AB - Enantiomers of 4'-aza-2',3'-dideoxynucleosides have been prepared by two different synthetic approaches, on the basis of 1,3-dipolar cycloaddition of a chiral nitrone. Cytotoxicity and apoptotic activity have been investigated. (5'S) 5-Fluoro-1-isoxazolidin-5-yl-1H-pyrimidine-2,4-dione [(-)-AdFU], while showing low level of cytotoxicity, is a good inductor of apoptosis on lymphoid and monocytoid cells, acting as a strong potentiator of Fas-induced cell death. PMID- 12904075 TI - A comparison of phosphonothioic acids with phosphonic acids as phosphatase inhibitors. AB - Phosphorothioates, analogues of phosphate esters in which a sulfur replaces an oxygen atom in the phosphoryl group, are competent surrogate substrates for a number of phosphatases. In some cases the thio analogues show similar binding (as estimated by K(m)) while other phosphatases show quite different K(m) values for phosphate compared to phosphorothioate esters. On this basis it was hypothesized that there might be different inhibitory tendencies by the nonhydrolyzable analogues, phosphonothioic acids compared with phosphonic acids. A series of phosphonothioic acids and corresponding phosphonic acids were synthesized and their inhibitory properties were compared toward human placental and E. coli alkaline phosphatases, the protein-tyrosine phosphatase from Yersinia, and the serine/threonine protein phosphatases PP2C and lambda. Sulfur substitution for oxygen gives the phosphonothioic acids pK(a) values that are close to those of phosphate esters, in contrast to the higher pK(a) values typical of phosphonic acids. Despite different steric requirements and differences in charge distribution in the anions of phosphonothioic acids compared with phosphonic acids, it was found that, with some exceptions, differences in inhibitory properties were modest. PMID- 12904077 TI - Structure-activity relationships of novel cyclic alpha-MSH/beta-MSH hybrid analogues that lead to potent and selective ligands for the human MC3R and human MC5R. AB - It has been shown by extensive studies that alpha-MSH bioactivity is critically dependent on the core or central tetrapeptide sequence, His-Phe-Arg-Trp, however with poor selectivity for the human MC3R-MC5R. The structure-activity relationships study here is aimed at identifying lead structures or templates of this core sequence by the use of different conformational constraints that might impart changes in its topography and thus promote differences in potency and selectivity at these receptors. Our peptide library consists of a novel series of cyclic alpha-MSH analogues that have disulfide bridges between Cys or Cys-like residues at positions 4 and 10, giving rise to 23-membered rings fused at the C terminal end with the C-terminal fragment of beta-MSH (Pro-Pro-Lys-Asp). While such constraints of the peptide backbone with disulfide bridges of different chirality affect potency and selectivity at these receptors, further changes in the hydrophobicity at position 7 with either a D-Phe or D-Nal(2') and replacement of a His with a Pro in position 6 cause additional effects. Thus, the most interesting lead compounds that emerged from this study are (1) compound 5, Ac c[Cys-Glu-His-D-Phe-Arg-Trp-D-Cys]-Pro-Pro-Lys-Asp-NH(2) (IC(50) = 10 nM), which is the first potent and highly selective antagonist ligand for the hMC5R (560 fold vs the MC3R and 1000-fold vs the MC4R); (2) compound 7, Ac-c[Cys-Glu-Pro-D Nal(2')-Arg-Trp-Cys]-Pro-Pro-Lys-Asp-NH(2) (IC(50) = 31 nM), which is a highly selective antagonist analogue for the MC3R (560-fold vs the hMC4R and about 3000 fold vs the hMC5R; and (3) compound 9, Ac-c[Pen-Glu-His-D-Nal(2')-Arg-Trp-Cys] Pro-Pro-Lys-Asp-NH(2) (IC(50) = 3 nM), which is more potent than 7 at the MC3R but not as selective. PMID- 12904076 TI - A novel class of nonpeptidic biaryl inhibitors of human cathepsin K. AB - A novel series of nonpeptidic biaryl compounds was identified as potent and reversible inhibitors of cathepsin K. The P2-P3 amide bond of a known amino acetonitrile dipeptide 1 was replaced with a phenyl ring, thereby giving rise to this biaryl series that retained potency vs cathepsin K and showed an improved selectivity profile against other cathepsins. Structural modification within this series resulted in the identification of compound (R)-2, a potent human cathepsin K inhibitor (IC(50) = 3 nM) that is selective versus cathepsins B (IC(50) = 3950 nM), L (IC(50) = 3725 nM), and S (IC(50) = 2010 nM). In an in vitro assay involving rabbit osteoclasts and bovine bone, compound (R)-2 inhibited bone resorption with an IC(50) of 95 nM. It was shown that, unlike some peptidic nitrile inhibitors of cysteine proteases, the nitrile moiety of (R)-2 is not converted to the corresponding amide 3 by cathepsin K. This indicates that this class of nonpeptidic nitrile inhibitors is unlikely to be hydrolyzed by cysteine proteases. Furthermore, the inhibition of cathepsin K by compound (R)-2 was shown to be fully reversible and not observably time-dependent. To demonstrate the efficacy of compound (R)-2 in vivo, it was administered to ovariectomized (OVX) rhesus monkeys at 20 mg/kg, po once daily for 8 days, and a urinary marker of bone turnover, N-telopeptide of type I collagen (uNTx), was measured. During the eight-day dosing period, the mean reduction by compound (R)-2 in uNTx was 80% (p < 0.001). This demonstrates that inhibition of cathepsin K leads to an inhibition of this bone resorption marker in OVX rhesus monkeys and strongly suggests that inhibition of cathepsin K is a viable therapeutic approach for the treatment of osteoporosis. PMID- 12904078 TI - Water soluble, core-modified porphyrins. 3. Synthesis, photophysical properties, and in vitro studies of photosensitization, uptake, and localization with carboxylic acid-substituted derivatives. AB - Water soluble, core-modified porphyrins 1-5 bearing 1-4 carboxylic acid groups were prepared and evaluated in vitro as photosensitizers for photodynamic therapy. The 21,23-core-modified porphyrins 1-5 gave band I absorption maxima with lambda(max) of 695-701 nm. The number of carboxylic acid groups in the dithiaporphyrins 1-4 had little effect on either absorption maxima (lambda(max) of 696-701 nm for band I) or quantum yields of singlet oxygen generation [phi((1)O(2)) of 0.74-0.80]. Substituting two Se atoms for S gave a shorter band I absorption maximum (lambda(max) of 695 nm) and a smaller value for the quantum yield for generation of singlet oxygen [phi((1)O(2)) of 0.30]. The phototoxicity of 1-5 was evaluated against R3230AC cells. The phototoxicities of dithiaporphyrin 2, sulfonated thiaporphyrin 30, HPPH, and Photofrin were also evaluated against Colo-26 cells in culture using 4 J cm(-2) of 570-800 nm light. Compound 2 was significantly more phototoxic than sulfonated dithiaporphyrin 30, HPPH, or Photofrin. Cellular uptake was much greater for compounds 1, 2, and 5 relative to compounds 3 and 4. Confocal scanning laser microscopy and double labeling experiments with rhodamine 123 suggested that the mitochondria were an important target for dithiaporphyrins 1 and 2. Inhibition of mitochondrial cytochrome c oxidase activity in whole R3230AC cells was observed in the dark with compounds 1 and 30 and both in the dark and in the light with core-modified porphyrin 2. PMID- 12904079 TI - Hydroxyurea analogues as kinetic and mechanistic probes of the nitric oxide producing reactions of hydroxyurea and oxyhemoglobin. AB - Derivatives of N-hydroxyurea that contain an N-hydroxy group react with oxyhemoglobin to form methemoglobin and variable amounts of nitrite/nitrate. Compounds with an unsubstituted -NHOH group produce the most nitrite/nitrate, which provides evidence for nitric oxide formation. The rate of reaction of these N-hydroxyurea derivatives with oxyhemoglobin correlates well with that compound's oxidation potential. Aromatic N-hydroxyureas react 25-80-fold faster with oxyhemoglobin than with N-hydroxyurea, suggesting other N-hydroxyurea analogues may be superior nitric oxide donors. Electron paramagnetic resonance spectroscopy shows that the formation of a low-spin methemoglobin-hydroxyurea complex is critical for iron nitrosyl hemoglobin formation. These results show that iron nitrosyl hemoglobin formation from the reaction of hydroxyureas and hemoglobin requires an unsubstituted -NHOH group and that the nitrogen atom of the non-N hydroxy group must contain at least a single hydrogen atom. These results should guide the development of new hydroxyurea-based nitric oxide donors and sickle cell disease therapies. PMID- 12904080 TI - Bispyridinium cyclophanes: novel templates for human choline kinase inhibitors. AB - The synthesis and biological activities of four novel bispyridinium cyclophanes as choline kinase (ChoK) inhibitors are presented. Their synthetic methodology has been optimized according to dilution, temperature, and reaction time and provides pure bispyridinium cyclophanes in high yields very easily. One of these cyclophanes (6, 4,8-diaza-3(1,4),9(4,1)-dipyridina-1(1,4),6(1,3) dibenzenacyclodecaphan-3(1),9(1)-bis(ilium) dibromide) has an IC(50(ChoK)) of 0.3 microM and is the most potent human ChoK inhibitor described to date. PMID- 12904081 TI - Synthesis and evaluation of pharmacological properties of novel annelated 2,3 benzodiazepine derivatives. AB - New cyclofunctionalized 2,3-benzodiazepines characterized by a triazolone or triazindione ring fused on the "c" edge of the heptatomic nucleus have been prepared. These compounds were evaluated as potential anticonvulsant agents, and some of them proved to be more potent noncompetitive 2-amino-3-(3-hydroxy-5 methylisoxazol-4-yl)propionate (AMPA) receptor antagonists. In particular, 8,9 dimethoxy-6-(4-bromophenyl)-11H-[1,2,4]triazolo[4,5-c][2,3]benzodiazepin-3(2H) one (5b) was almost 10-fold more active than GYKI-52466 and 3.5-fold more active than Talampanel. Furthermore, 5b potently reduced AMPA-evoked currents in electrophysiological experiments. PMID- 12904082 TI - New potential uroselective NO-donor alpha1-antagonists. AB - A recent uroselective alpha(1)-adrenoceptor antagonist, REC15/2739, has been joined with nitrooxy and furoxan NO-donor moieties to give new NO-donor alpha(1) antagonists. All the compounds studied proved to be potent and selective ligands of human cloned alpha(1a)-receptor subtype. Derivatives 6 and 7 were able to relax the prostatic portion of rat vas deferens contracted by (-)-noradrenaline because of both their alpha(1A)-antagonist and their NO-donor properties. PMID- 12904083 TI - Conformationally defined retinoic acid analogues. 5. Large-scale synthesis and mammary cancer chemopreventive activity for (2E,4E,6Z,8E)-8-(3',4'-dihydro 1'(2'H)-naphthalen-1'-ylidene)-3,7-dimethyl-2,4,6-octatrienoic acid (9cUAB30). AB - Retinoids that activate the nuclear retinoid X receptors (RXRs) display potential for chemoprevention of breast cancer. We previously reported that 9cUAB30 (1) is an RXR-selective retinoid. To explore its in vivo chemopreventive activity, multigram quantities of 1 were needed. Here, we describe a modified synthesis that yields up to 100 g of 1. We further demonstrate that 1 is very effective in the prevention of N-methyl-N-nitrosourea induced mammary cancers in rats without signs of toxicity. PMID- 12904086 TI - Atazanavir. AB - Atazanavir is a novel azapeptide protease inhibitor with high specificity for, and activity against, HIV-1 protease. The resistance profile of atazanavir is distinct, with an I50 L protease substitution appearing to be the signature mutation. Atazanavir was not associated with increases in total cholesterol, low density lipoprotein-cholesterol or triglyceride levels after 108 weeks. Atazanavir has a pharmacokinetic profile that allows for once-daily oral administration. It is a moderate inhibitor of hepatic cytochrome P450 enzymes and interacts with several drugs. In combination with stavudine plus didanosine, atazanavir 200, 400 or 500 mg once daily produced a rapid and sustained reduction from baseline in viral load of 2.57, 2.42 and 2.53 log(10) copies/mL, respectively, in treatment-naive patients after 48 weeks, compared with a decrease of 2.33 log(10) copies/mL with nelfinavir 750 mg three times daily. Nausea was the most clinically relevant adverse event reported in patients receiving atazanavir-based regimens. PMID- 12904084 TI - Breakthroughs in the management of multiple myeloma. AB - Although multiple myeloma remains a terminal illness, the past four decades have seen a dramatic change in the outlook for a newly diagnosed patient in terms of therapies available, supportive care and insight into the pathogenesis of this disease. Among the newer agents available for treatment, thalidomide has been resurrected and discovered to be a valuable therapy for myeloma. Thalidomide appears to work, at least in part, through its anti-angiogenic properties, but much remains to be learned about its mechanism of action as well as optimal administration regimens. With the development of increasingly more potent bisphosphonates it has become possible to diminish the painful skeletal complications of myeloma, one of the most devastating problems of this disease. The most recent generation of bisphosphonates, pamidronic acid and zoledronic acid, have provided a statistically significant decrease in the skeletal complications of myeloma when used in a prophylactic manner. These agents appear to work by inhibiting osteoclast function. Progressive improvement in cytogenetic techniques has now demonstrated that almost all patients with myeloma have chromosomal abnormalities, some of which appear to confer varying degrees of prognostic significance. In particular, the changes in chromosome 13 are associated with an unusually poor outcome. These findings are serving as a guide toward learning more about the pathogenesis of myeloma as well as in identifying potential targets for therapy. Stem cell transplantation has emerged as the standard treatment for the large majority of patients with myeloma following the demonstration of superior complete remission and survival, both disease-free and overall, in a French randomised trial. Unfortunately, virtually all patients will eventually relapse following autologous stem cell transplantation, prompting continuing efforts such as tandem transplants, CD34+ selection, as well as modifications in the conditioning regimen to improve outcomes. Allogeneic bone marrow transplants appear to offer a better chance for a possible cure of myeloma but have been associated with an unusually high mortality. However, this approach is being revived with the advent of the less toxic non-myeloablative transplant that has provided an 81% short-term survival in a trial combining this approach with an initial conventional autologous bone marrow transplant. Immunotherapy with dendritic cells appears now to be a feasible way to enhance innate or acquired immunity to help eliminate minimal residual disease following autologous bone marrow transplant. Unfortunately, a cure for myeloma remains elusive but the continuing advances in management may significantly prolong survival in affected patients. PMID- 12904091 TI - The demise of HRT? The long-term safety of hormone replacement therapy. AB - The role of hormone replacement therapy (HRT) in the health of middle-aged women has come a full circle. HRT has been widely accepted as the treatment of choice for the management of menopausal symptoms. However, the Women's Health Initiative (WHI) and other recent randomised controlled trials have failed to confirm beliefs of other potential benefits in reducing the risk of coronary artery disease (CAD) and stroke. Indeed, early increases in cardiac event and stroke rate have been seen in women taking combination HRT. An increased risk of breast cancer diagnosis has also been confirmed in HRT users. The use of HRT now needs to be regarded as a short-term therapy for menopausal symptom management with treatment individualised for each woman. PMID- 12904092 TI - Safety and efficacy of disease-modifying antirheumatic agents in rheumatoid arthritis and juvenile rheumatoid arthritis. AB - The definition of disease-modifying antirheumatic drugs (DMARDs) has changed dramatically over the last decade. Current expectations of efficacy now include amelioration of signs and symptoms of disease activity as well as slowing, if not complete cessation, of disease progression as evidenced by Xray progression and significant improvement of patient function. Rheumatologists assess the safety profile of these agents more critically in an attempt to increase the risk:benefit profile. Traditional agents, such as methotrexate (MTX), sulfasalazine and leflunomide have provided patients with substantial relief of symptoms and some decrease of X-ray progression but have been hampered by the frequent occurrence of significant adverse events (AEs) and inability to maintain benefit for a prolonged period of time. With the increased understanding of the basic mechanism of the disease process, there has been the introduction of four biological disease-modifying agents introduced into clinical practice which have substantially increased the risk:benefit ratio for patients with various rheumatic diseases. PMID- 12904094 TI - Anticonvulsant-induced dyskinesia. AB - Anticonvulsant-induced dyskinesia (AID) is an underdiagnosed side effect of many anticonvulsants that may take place during initial or chronic treatment at normal or toxic drug levels. The occurrence of AID subjects the patient to another medical condition and may prompt an extensive work-up. Similarities with other drug-induced dyskinesias and some animal studies suggest that dopaminergic dysfunction in the basal ganglia is pivotal in the occurrence of dyskinesia. Clinical presentation and outcomes are variable; however, in most cases, dyskinesias respond well to anticonvulsant withdrawal. Enhancing the awareness of AID is important in light of the recent development of many new anticonvulsants and their wider clinical use. PMID- 12904093 TI - Safety of antidepressants in the elderly. AB - Until the 1980s, the two major classes of antidepressants, the tricyclics and the monoamine oxidase inhibitors (MAOIs), were effective but had severe side effects, requiring monitoring by psychiatrists. The past several years have brought new classes of antidepressants that are safer for the patient to take and far easier for the non-psychiatrist to prescribe. Whilst this is of enormous value, it leaves the physician with the dilemma of which one to prescribe. These new antidepressants cannot safely be used interchangeably. This paper will discuss each of the antidepressants presently available, with particular emphasis on safety in the elderly. Drug interactions, side effects and particular challenges to the older patient will be described. The authors will then advise a general strategy for prescribing antidepressants. PMID- 12904089 TI - Carvedilol: a review of its use in chronic heart failure. AB - Carvedilol (Dilatrend) blocks beta(1)-, beta(2)- and alpha(1)-adrenoceptors, and has antioxidant and antiproliferative effects. Carvedilol improved left ventricular ejection fraction (LVEF) in patients with chronic heart failure (CHF) in numerous studies. Moreover, significantly greater increases from baseline in LVEF were seen with carvedilol than with metoprolol in a double-blind, randomised study and in a meta-analysis. Carvedilol also reversed or attenuated left ventricular remodelling in patients with CHF and in those with left ventricular dysfunction after acute myocardial infarction (MI). Combined analysis of studies in the US Carvedilol Heart Failure Trials Program (patients had varying severities of CHF; n = 1094) revealed that mortality was significantly lower in carvedilol than in placebo recipients. In addition, the risk of hospitalisation for any cardiovascular cause was significantly lower with carvedilol than with placebo. Mortality was significantly lower with carvedilol than with metoprolol in patients with mild to severe CHF in the Carvedilol Or Metoprolol European Trial (COMET) [n = 3029]. The Carvedilol Prospective Randomised Cumulative Survival (COPERNICUS) trial (n = 2289) demonstrated that compared with placebo, carvedilol was associated with significant reductions in all-cause mortality and the combined endpoint of death or hospitalisation for any reason in severe CHF. All-cause mortality was reduced in patients who received carvedilol in addition to conventional therapy compared with those who received placebo plus conventional therapy in the Carvedilol Post-Infarct Survival Control in LV Dysfunction (CAPRICORN) trial (enrolling 1959 patients with left ventricular dysfunction following acute MI). Carvedilol was generally well tolerated in patients with CHF. Adverse events associated with the alpha- and beta-blocking effects of the drug occurred more commonly with carvedilol than with placebo, whereas placebo recipients were more likely to experience worsening heart failure. In conclusion, carvedilol blocks beta(1)-, beta(2)- and alpha(1) adrenoceptors and has a unique pharmacological profile. It is thought that additional properties of carvedilol (e.g. antioxidant and antiproliferative effects) contribute to its beneficial effects in CHF. Carvedilol improves ventricular function and reduces mortality and morbidity in patients with mild to severe CHF, and should be considered a standard treatment option in this setting. Administering carvedilol in addition to conventional therapy reduces mortality and attenuates myocardial remodelling in patients with left ventricular dysfunction following acute MI. Moreover, mortality was significantly lower with carvedilol than with metoprolol in patients with mild to severe CHF, suggesting that carvedilol may be the preferred beta-blocker. PMID- 12904090 TI - Insulin glargine: an updated review of its use in the management of diabetes mellitus. AB - Insulin glargine is a human insulin analogue prepared by recombinant DNA technology. Modification of the human insulin molecule at position A21 and at the C-terminus of the B-chain results in the formation of a stable compound that is soluble at pH 4.0, but forms amorphous microprecipitates in subcutaneous tissue from which small amounts of insulin glargine are gradually released. The plasma concentration versus time profile of insulin glargine is therefore relatively constant in relation to conventional human insulins, with no pronounced peak over 24 hours. This allows once-daily administration as basal therapy. Early randomised trials with insulin glargine generally showed greater reductions in fasting blood or plasma glucose levels and a reduced frequency of nocturnal hypoglycaemia relative to neutral protamine Hagedorn (NPH) insulin in patients with type 1 diabetes mellitus. In addition to this basal therapy, patients continued to use the regular mealtime insulin regimen to which they were accustomed. More recent data with insulin glargine have included evidence of improved glycaemic control, with improvements in satisfaction with treatment over NPH insulin. Furthermore, the time of day at which insulin glargine is injected has no clinically relevant effect on glycaemic control in these patients. There are also data from small, nonblind studies to suggest comparable glycaemic control with insulin glargine and continuous subcutaneous insulin infusion. Results from comparative studies and meta-analyses in individuals with type 2 diabetes show lower incidences of nocturnal hypoglycaemia with insulin glargine than with NPH insulin, with two studies showing a significantly greater improvement in glycosylated haemoglobin levels with insulin glargine than with NPH. Insulin glargine is well tolerated, and is not associated with greater immunogenicity or increases in bodyweight than NPH insulin. Long-term data show maintenance of glycaemic control with insulin glargine for up to 39 months in adults and children with type 1 and adults with type 2 diabetes. In conclusion, insulin glargine is an effective and well tolerated basal insulin therapy when given as a single daily subcutaneous injection to patients with diabetes, with benefits in terms of glycaemic control and reduced frequency of hypoglycaemia over regimens based on conventional basal insulins. Accumulating data and official recommendations show the suitability of insulin glargine for first-line use in selected patients with type 2 diabetes who require insulin treatment, as well as in patients with type 1 disease, and confirm its use in children and adolescents. PMID- 12904096 TI - Drug-induced vasculitis. AB - Therapeutic agents from virtually every pharmacological class have been implicated in the development of drug-induced vasculitis. Clinical manifestations range from small vessel hypersensitivity vasculitis and leukocytoclastic vasculitis to clinical syndromes indistinguishable from classical systemic forms of vasculitis such as Wegener's granulomatosis, polyarteritis nodosa and Churg Strauss syndrome (CSS). The exact pathogenic mechanisms involved remain to be elucidated; but both cell-mediated and humoral immunity appear to play important roles. Affected individuals may present with cutaneous involvement alone or life threatening systemic involvement, which may result in severe and sometimes fatal illness. Since clinical presentation as well as serological and pathological parameters is identical to idiopathic forms of vasculitis, a high index of suspicion is necessary to accurately and expeditiously diagnose drug-induced vasculitis. Withdrawal of the offending agent alone is often sufficient to induce prompt resolution of clinical manifestations, obviating the need for systemic corticosteroids or more powerful forms of immunosuppression. PMID- 12904095 TI - Haematological safety of antipsychotic drugs. AB - Haematological abnormalities are frequently encountered during treatment with antipsychotic drugs. Most of these are mild and of no clinical significance. In the case of many, there is often difficulty in establishing a cause-and-effect relationship between the drug and the abnormality. However, in a small minority of patients, hazardous, potentially life-threatening haematological effects can occur due to a combination of pharmacological and host factors. These include leucopenia and agranulocytosis. Although such effects are rare, it is essential that they are diagnosed and managed promptly. In this paper, the authors review the haematological adverse effects and safety of antipsychotic drugs and present a strategy for prevention. PMID- 12904085 TI - Management of trigeminal autonomic cephalgias and hemicrania continua. AB - The trigeminal autonomic cephalgias (TACs) are a group of primary headache disorders characterised by unilateral trigeminal distribution pain that occurs in association with ipsilateral cranial autonomic features. This group of headache disorders includes cluster headache, paroxysmal hemicrania and short-lasting unilateral neuralgiform headache attacks with conjunctival injection and tearing (SUNCT syndrome). Although hemicrania continua has previously been classified amongst the TACs, its nosological status remains unclear. Despite their similarities, these disorders differ in their clinical manifestations and response to therapy, thus underpinning the importance of recognising them. We have outlined the clinical manifestations, differential diagnoses, diagnostic workup and the treatment options for each of these syndromes. PMID- 12904097 TI - Ocular adverse drug reactions. AB - Drug-induced ocular side effects are described according to recent reports from the literature, the National Registry of Drug-Induced Ocular Side Effects, the World Health Organization (WHO) and the FDA. Adverse events are categorised as certain, probable/likely, possible, unlikely, conditional/unclassified and unassessable/unclassifiable where indicated. Ocular side effects of clinical importance are highlighted with guidelines for recognition, reporting and treatment of adverse drug reactions (ADRs). The current and future status of pharmacovigilance in ocular toxicology is addressed. PMID- 12904098 TI - Gastrointestinal side effects of drugs. AB - Drugs can have adverse effects on any part of the gastrointestinal (GI) tract from mouth to colon. It is essential that a detailed and accurate drug history is taken in patients presenting with GI complaints. Many drug-induced effects will regress or heal on cessation of treatment. NSAIDs are usually associated with gastric and duodenal ulcers but are also recognised to cause lichen planus in the mouth, oesophageal inflammation and strictures, and small bowel and colonic ulcers and strictures. A newer class of anti-inflammatory drugs, the cyclooxygenase-2 (COX-2)-selective inhibitors, have been developed and have a more favourable GI safety profile than standard NSAIDs. Acute diarrhoea, relapse of inflammatory bowel disease (IBD), microscopic colitis and acute pancreatitis are also induced by ingestion of standard NSAIDs. The calcium antagonists, phenytoin and cyclosporin, induce gum hyperplasia, particularly in patients with poor oral hygiene. Alendronate, a bisphosphonate, has been associated with development of oesophageal ulcers, and specific recommendations are now given to reduce this complication. Of the many different forms of colitis associated with drug ingestion, the most frequent is pseudomembranous colitis. This is a complication of antibiotics and is caused by the toxin produced by Clostridium difficile. Many drugs have been associated with the development of acute pancreatitis, although a definite cause and effect relationship has been shown for only a few drugs. These include didanosine, furosomide, corticosteroids, azathioprine and sodium valproate. PMID- 12904099 TI - The Cambridge Healthtech Institute's 9th Annual Conference on Blood Safety. AB - This article summarises, and provides a personal commentary on, the Cambridge HealthTech Institute's Conference on Blood Safety in February 2003. Most of the converging areas which contribute to this field were amply covered at the conference by speakers of a high calibre and international standing. PMID- 12904100 TI - Are drugs really toxic for older people? AB - Many diseases are now treatable with modern drugs. Elderly people, because they suffer from age-related illnesses, stand to gain the most, but they are also at risk from adverse drug reactions (ADRs). There are complex reasons for the increased frequency of ADRs, including poor prescribing, polypharmacy, altered drug handling and response, and poor compliance. Difficulties with interpretation of apparent ADR data include uncertainties about exposure and various confounding factors, for example, confounding by indication. Some simple prescribing guidelines can significantly help to minimise the extent of the clinical problem. These include: PMID- 12904101 TI - The overall safety profile of currently available vaccines directed against infectious diseases. AB - The vaccines currently used worldwide for the prevention of infectious diseases are quite safe in comparison to most pharmaceutical and biological agents. Vaccine components may contribute to inflammatory, allergic or anaphylactic reactions. Most adverse events are transient and well-tolerated. Transient severe adverse reactions occur at rates of one in one thousand vaccinations; permanent severe adverse reactions occur on the order of one in one million. The most common of the severe adverse reactions are syncope and allergic reactions. Providers can take steps to prevent or ameliorate these reactions by pursuing both prelicensure testing (albeit limited) and postlicensure testing and monitoring. Systems that enhance the detection of safety concerns include national passive and active surveillance as well as regional vaccine registries and provider-based patient education. Since vaccines are used in universal programmes, their safety is paramount to their continued acceptance. Healthcare managers, including administrators of hospitals, clinics, practice groups, health maintenance organisations (HMOs) and managed care plans, can and should support providers in minimising adverse events associated with vaccines by supporting postvaccination observation policies, postlicensure testing and surveillance, vaccine registries and patient education systems. PMID- 12904102 TI - Safety of immunisation and adverse events following vaccination against hepatitis B. AB - Hepatitis B vaccines (HBVs) are composed of highly purified preparations of hepatitis B virus surface antigen (HBsAg). An adjuvant, either aluminium phosphate or aluminium hydroxide, is added to the vaccines, which are sometimes preserved with thiomersal. In placebo-controlled studies, common side effects other than local reactions were reported no more frequently among vaccine recipients than among individuals receiving a placebo. A number of controversial adverse events have, however, been purported to be associated with HBVs, including rheumatoid arthritis (RA), diabetes, demyelinating diseases (e.g., multiple sclerosis [MS]), chronic fatigue syndrome, and more recently, lymphoblastic leukaemia. In addition, the safety of the thiomersal and aluminium contained in the vaccine has also been under close scrutiny. These issues have been reviewed by a number of country-specific or international independent review committees such as that of the US Institute of Medicine (IOM) and the World Health Organization's (WHO) Global Advisory Committee on Vaccine Safety (GACVS). Upon review of the scientific evidence, none of the serious allegations have so far been confirmed. On the contrary, scientific evidence has accumulated to disprove many of the allegations. In particular, the IOM committee has concluded that the evidence favoured rejection of a causal relationship between HBV administered to adults and incident MS or MS relapse. Whilst it is important to continue monitoring some of the safety issues, there is no evidence to suggest that the WHO should consider altering its recommendation that all countries should have universal infant and/or adolescent immunisation programmes. The risks of hepatitis B vaccination are only theoretical in comparison with clear benefits in terms of cirrhosis and cancer prevention, and the HBV remains one with an excellent safety profile. PMID- 12904103 TI - Risk of seizures associated with psychotropic medications: emphasis on new drugs and new findings. AB - Psychotropic medications in the classes of antidepressants, antipsychotics and mood stabilisers have been recognised in the literature and clinical settings as having high epileptogenic potential. Among these three classes, clozapine, tricyclic antidepressants (TCAs) and lithium are agents that clinicians have historically recognised as precipitants of drug-induced seizures. There are few reports that review the epileptogenic risk of newer psychotropic agents; in this qualitative review, the authors provide an update on the most recently published reports on seizures associated with antidepressants, antipsychotics, mood stabilisers, anxiolytics and sedative-hypnotics. In general, the epileptogenic risks of the newer psychotropic agents appear to be quite low as long as dosing strategies are consistent with recommended guidelines. Whilst newer psychotropic medications appear to be safe in patients with epilepsy, few studies have specifically addressed this population. In addition, the potential for drug interactions between antiepileptic drugs and psychotropics may be substantial with certain agents. For example, many psychotropes are both substrates and inhibitors of cytochrome P450 (CYP450) isoenzymes, whilst many antiepileptic drugs are both substrates and inducers of CYP450 activity. Every attempt should be made to minimise potential interactions when these agents are concomitantly administered. PMID- 12904104 TI - Antidepressant-induced hepatotoxicity. AB - Depression is a chronic, severe and increasingly prevalent illness associated with substantial morbidity, mortality and healthcare costs. Antidepressant drugs, the cornerstone of depression treatment, are not devoid of adverse effects, including hepatotoxicity. To review the risk of liver toxicity related to major antidepressants, the authors have followed structural criteria focusing on the underlying mechanism presumably involved and the role of particular chemical structures. The clinicopathological expression goes from transient increases in liver enzymes to fulminant liver failure. Classical antidepressants such as monoamine oxidase inhibitors (MAOIs) or tricyclic antidepressants (TCAs) seem to have the highest potential to induce liver damage compared with the newer drugs such as selective serotonin re-uptake inhibitors (SSRIs). The potential for severe hepatotoxicity associated with nefazodone is stressed. Guidelines for therapy and prevention of antidepressant-induced hepatotoxicity are also discussed. PMID- 12904105 TI - Hepatotoxic profile of catechol-O-methyltransferase inhibitors in Parkinson's disease. AB - Entacapone and tolcapone are selective catechol-O-methyltransferase (COMT) inhibitors developed recently as adjuncts to levodopa for the treatment of Parkinson's disease (PD). They extend the duration of action of levodopa. As a result, they increase 'on' time, decrease 'off' time and improve motor scores in patients with motor fluctuations. Both benefits and main side effects are related to increased dopaminergic activity. This paper reviews the use of those COMT inhibitors in PD with particular focus on the issue of hepatotoxicity. Neither tolcapone nor entacapone caused hepatotoxicity in preclinical studies. However, in 1998, four patients who were using tolcapone presented with serious liver dysfunction; three of them died due to acute liver failure. Tolcapone is now known to have the potential to cause hepatotoxicity in clinical use and experimental studies. It is now recommended that tolcapone be administered only in patients with motor fluctuations who are no longer satisfactorily treated with other medications for PD. Routine liver monitoring is now mandatory with this agent. Entacapone has been described as a well-tolerated and safe drug in recent experimental studies, human clinical trials and postmarketing surveillance. It can be offered to any patient with motor fluctuations and routine liver monitoring is not required. PMID- 12904106 TI - Combination lipid-lowering therapy with statins: safety issues in the postcerivastatin era. AB - Combination lipid-altering regimens represent an emerging clinical paradigm to meet increasingly stringent consensus lipoprotein targets for coronary prevention. This practice, together with escalating prevalences of coronary artery disease in certain ageing (western industrial) populations, polypharmacy in the elderly and the recent voluntary market withdrawal of cerivastatin, warrants a re-examination of the safety profiles of 3-hydroxy-3-methylglutaryl co enzyme A (HMG-CoA) reductase inhibitors (i.e., statins). These agents are exceedingly well-tolerated in the vast majority of patients, very infrequently precipitating musculoskeletal symptoms and/or signs. Statins vary in their pharmacological profiles, leading to distinct levels of systemic exposure and capacities to penetrate skeletal myocytes. Pharmacokinetic interactions with certain agents increase the likelihood of statin-induced myopathy and, in exceedingly rare instances, potentially fatal rhabdomyolysis with myoglobinuria and renal failure. As with other medical decisions, the anticipated benefits of long-term statin therapy, with or without other lipid-altering agents, need to be weighed against the prospects of clinically significant drug interactions. In clinical trials and postmarketing surveillance, the two statins that are not metabolised by the cytochrome P450 3A4 system (fluvastatin and pravastatin) have exhibited very low propensities to elicit myopathy when combined with other agents. These agents should be considered initially when contemplating combination lipid-lowering regimens for coronary prevention. PMID- 12904107 TI - Drug-induced cholestasis. AB - Drugs may cause several overlapping syndromes of cholestasis, the pathophysiological syndrome resulting from impaired bile flow. These reactions comprise approximately 17% of all hepatic adverse drug reactions (ADRs) and they may be severe. Causes of 'pure' (bland) cholestasis include oestrogens and anabolic steroids; rarer associations are with antimicrobials and NSAIDs. 'Cholestatic hepatitis' is a common drug reaction in which liver injury and inflammation cause significant elevation of serum alanine aminotransferase (ALT) as well as cholestasis. Chlorpromazine and ketoconazole are classic examples, but it is now exemplified by amoxycillin-clavulanate and other oxy-penicillins. Chronic cholestasis results from small bile duct injury leading to the vanishing bile duct syndrome (VBDS), a disorder mimicking primary biliary cirrhosis, or from injury to larger bile ducts causing secondary sclerosing cholangitis. Whilst there is increasing evidence of a genetic predisposition to cholestatic drug reactions, there are currently no pretreatment tests to predict drug safety. Prevention of severe reactions therefore relies on early detection of liver injury and prompt drug withdrawal. Symptomatic management includes relief of pruritus and correction of fat-soluble vitamin deficiency. In small cohort studies, ursodeoxycholic acid (UDCA) arrested progressive cholestasis in two thirds of cases, but evidence for use of corticosteroids is anecdotal. This review considers diagnosis, pathogenesis, prevention and management of drug induced cholestasis, with particular reference to frequently- and newly-described causes. PMID- 12904108 TI - Anticipating and treating opioid-associated adverse effects. AB - Opioids are frequently avoided as viable tools in the management of pain due to perceived dangerous or untoward adverse drug events. Whilst they are relatively safe options for the treatment of pain, side effects and toxicities do exist and should be anticipated by the provider. The central nervous, gastrointestinal, genito-urinary, integumentary, metabolic/endocrine, cardiovascular, pulmonary, hepatic/renal, ocular and immune systems all manifest changes associated with opioid therapy. These adverse events, ranging from nuisance to therapy-limiting, are manageable when addressed quickly and appropriately. Opioids are safe and efficacious analgesics when these effects are considered. PMID- 12904109 TI - Risk-benefit effects of implantable contraceptives in women. AB - Contraceptive implant technology has been used by millions of women throughout the world. The three marketed implant systems today are levonorgestrel-releasing implants: Norplant and Jadelle, and a single etonogestrel-releasing implant, Implanon. The main benefits common to these delivery systems are their safety, high effectiveness, ease of use, long duration of action (3 - 5 years) and reversibility. Bleeding disturbances are the main adverse events associated with implantable contraceptives. Other minor risks relate to the insertion and removal procedures, which require adequately trained providers as well as aseptic techniques. Furthermore, since initiation and discontinuation of use is provider dependent and not controlled by the user, there may be a risk of coercion of use on the one hand or, on the other hand, difficulty in access to initiating use, if trained providers are not readily available. Although no single contraceptive method is perfect or appealing to all, contraceptive implants are safe and fulfil a very important need among fertility regulation methods. PMID- 12904110 TI - Safety of medicines in children. AB - Unlicensed and off-label prescribing is common in children. This is likely to be due to a combination of the difficulty in carrying out reliable clinical trials in children and the lack of financial reward for the industry. The term 'paediatric' encompasses preterm babies to near adult-sized adolescents, and all aspects of drug handling differ over this age range, making extrapolation very problematic. Recently, adrenal failure with inhaled corticosteroid use in children has been highlighted. Examination reveals that this is due to a combination of factors common in paediatric prescribing; including the assumption that adult diseases are the same as paediatric diseases, the lack of good diagnostic tools in small children and lack of thought about problems of drug delivery. Although some steps are being taken to improve paediatric prescribing, much more needs to be done. One approach may be to offer incentives to the pharmaceutical industry. The present state of affairs is unsatisfactory and cannot be allowed to continue. PMID- 12904111 TI - A review of hepatitis B vaccination. AB - Hepatitis B is one of the most important infectious causes of acute and chronic liver disease both in the US and worldwide. In order to combat the life threatening effects of hepatitis B infection, recombinant hepatitis B vaccines have been developed. The medical and scientific communities have generally accepted that recombinant hepatitis B vaccine - a highly purified, genetically engineered, single antigen vaccine - is a safe vaccine. Information is presented showing that hepatitis B vaccine contains yeast, aluminium, thimerosal and hepatitis B surface antigen epitopes, which may result in hepatitis B vaccine being associated with autoimmune diseases among susceptible adult vaccine recipients. There is little doubt that the benefits of this vaccine overall far outweigh its risks. Physicians and patients should evaluate the risks and benefits of hepatitis B vaccination and, together, make an informed consent decision as to whether to undergo vaccination. Individuals who experience an adverse reaction to hepatitis B vaccination should report it to the Vaccine Adverse Event Reporting System database and be advised that they may be eligible for compensation from the no-fault National Vaccine Injury Compensation Program, administered by the US Court of Claims. The authors strongly urge that additional research be conducted into the molecular basis of adverse events following hepatitis B vaccine administration, so that further recommendations may be made on how to improve their safety profiles. PMID- 12904112 TI - Safety profile of the triptans. AB - The triptans are 5-HT(1B/1D) agonists used as migraine and cluster-specific agents. Seven are in commercial use worldwide; in order of release these are sumatriptan, zolmitriptan, rizatriptan, naratriptan, almotriptan, frovatriptan and eletriptan. Sumatriptan has been in clinical use since 1991, and although postmarketing studies have stimulated much debate of triptan strengths and weaknesses, their overall safety profile appears excellent. The most serious adverse events are cardiovascular, due to coronary artery narrowing as a consequence of coronary artery 5-HT(1B) receptor activity. Triptans are contraindicated in patients with vascular disease. Other events are even more rare, and include the potential for drug-drug interactions, based on metabolic elimination pathways. Serotonin syndrome has been a concern, but one large prospective study failed to document a single case, and reports are sporadic and not clearly causative. PMID- 12904113 TI - Safety of short-acting nifedipine in children with severe hypertension. AB - Short-acting nifedipine has been abandoned as a treatment for severe hypertension in adults as a result of significant adverse effects. Despite this, it remains a popular choice for the treatment of severe hypertension in children. However, recent publications describing adverse effects of short-acting nifedipine in children similar to those reported in adults, have prompted some experts to question the continued use of this agent in children. In this review, available data on the pharmacology, clinical efficacy and safety of short-acting nifedipine are reviewed, and the advisability of using short-acting nifedipine is reassessed. Although low (< 0.25 mg/kg) doses of short-acting nifedipine may be safe in some hypertensive children, alternative agents that produce more controlled reductions in blood pressure, and that are easier to accurately dose and administer, should probably be chosen for the majority of children with severe hypertension. PMID- 12904114 TI - Management of toxicities associated with the administration of taxanes. AB - The taxanes (paclitaxel and docetaxel) are highly active cytotoxic antineoplastic agents. Common toxicities of the drugs include total alopecia, hypersensitivity reactions, bone marrow suppression (principally neutropenia), arthralgia, myalgias, and peripheral neuropathy. When administered as a 3-h infusion, paclitaxel appears to be associated with a lower risk of neutropenia and a greater risk of peripheral neuropathy, compared to either 24-h infusion paclitaxel or docetaxel (1-h infusion). Neither paclitaxel nor docetaxel is associated with a high risk for significant emesis. High cumulative doses of docetaxel have been shown to produce fluid retention (e.g., oedema, ascites, pleural effusions), while paclitaxel, when combined with doxorubicin, increases the risk of anthracycline-induced heart failure. Both paclitaxel and docetaxel have been administered at lower dose levels, on a weekly schedule, with acceptable toxicity profiles. In general, the side effects of the taxanes are manageable, and few patients discontinue treatment due to excessive toxicity. PMID- 12904115 TI - The role of investigative molecular toxicology in early stage drug development. AB - Molecular toxicology, the application of molecular biology principles and technologies to preclinical safety assessment, represents a key tool for understanding mechanisms of toxicity and assessing the risks associated with specific toxicities. The application of gene expression markers to early stage preclinical safety assessment has the potential to impact pipelines in two main areas: lead optimisation and issue management. Lead optimisation focuses on deprioritising leads with significant, development-limiting toxicological liabilities while advancing those compounds with the greatest chance of successfully navigating the gauntlet of preclinical and clinical safety studies. Issue management utilises mechanistic toxicology studies to position non development-limiting findings prior to the onset of Good Laboratory Practice studies in full development, and can help to identify and validate gene expression markers predictive of adverse events to avoid issues in second generation projects. In this review, the authors describe the application of molecular toxicology to a standard pharmaceutical testing funnel, provide examples of the successful application of gene expression markers, and discuss the potential for future impact in several broad categories of clinically relevant toxicity. PMID- 12904116 TI - Treatment of premenstrual dysphoria with selective serotonin re-uptake inhibitors: focus on safety. AB - Many women experience physical or mood symptoms associated with the menstrual cycle. For approximately 3 - 8% of women, the symptoms are severe enough to significantly affect social, domestic and occupational functioning. This cluster of primarily emotional and behavioural symptoms is now labelled premenstrual dysphoric disorder (PMDD). Women who meet criteria for PMDD do not usually respond to conservative interventions; selective serotonin re-uptake inhibitors (SSRIs) taken either daily or intermittently are considered to be an effective first-line therapy for this population. In this paper, the authors report on the efficacy and tolerability of SSRIs that are currently recognised as the treatment of choice for PMDD. PMID- 12904117 TI - Adverse effects of sedatives in children. AB - The application of sedation/analgesia in paediatric patients is rapidly expanding as less invasive, non-operative techniques of diagnosis and treatment are applied to the paediatric population. Medical providers who are asked to provide sedation may include radiologists, paediatricians, nurses and emergency physicians, as well as anaesthesiologists and intensive care physicians. At the same time, the range of drugs used in these settings has expanded considerably. As there is no single drug fulfilling the criteria for the ideal sedative (rapid-onset, rapid recovery, no adverse effects, immobility appropriate to procedure being performed), multiple drugs may be used in combination. It is imperative that practitioners using drugs for sedation/analgesia in children be aware of the adverse effect profile(s) of these drugs, both individually and in combination. The purpose of this review is to describe the adverse effects of sedative and reversal agents currently used in paediatric sedation/analgesia. PMID- 12904118 TI - Electronic Reporting for Clinical Drug Safety and Pharmacovigilance. 29 - 30 October 2002, Amsterdam, The Netherlands. AB - The 31 January 2003 deadline for electronic reporting to the European Agency for the Evaluation of Medicinal Products has helped to focus industry and regulator attention alike. This conference provided a wide perspective on background and scope, recent successes, and possible future developments by specialists from regulators, pharmaceutical industries, software vendors and more. Organised by IIR Conferences Ltd, attendees had the opportunity to gain an overview of all major aspects of electronic reporting of individual case safety reports, to broaden their knowledge, and to network with individuals concerned by these issues. PMID- 12904119 TI - 3rd International Symposium on Early Toxicity Screening. 11 December 2002, Philadelphia, Pennsylvania, USA. AB - The 3rd International Symposium on Early Toxicity Screening: Early ADME-Tox Screening Approaches included presentations by primarily non-academic scientists, including biotechnology companies, pharmaceutical scientists and contract research service leaders. A central theme heard throughout the presentations involved the fundamental objectives of and obstacles to ADME and toxicity testing during drug development. In summary, successful preclinical drug safety assessment involves understanding a model's limited ability to mimic the human drug response, increasing the model's ability to mimic the human drug response, and adopting new strategies that model better human drug responses, while simultaneously conducting safety assessment more quickly, more thoroughly and more cost effectively. PMID- 12904120 TI - Monitoring self-medication. AB - Self-medication, using non-prescription drugs, is beneficial to patients, healthcare professionals, the pharmaceutical industry and governments. Although many non-prescription medications are effective, misuse and abuse of these products can arise. Monitoring of specific populations and products, in addition to data recording and educating and informing patients, may go some way towards promoting safe and effective self-medication. PMID- 12904121 TI - Safety and efficacy of meningococcal group C conjugate vaccines. AB - The UK was the first country to implement a universal vaccination programme with conjugate polysaccharide vaccines against Neisseria meningitidis group C. This article reviews the pre- and postlicensure data on their efficacy and safety 3 years after the introduction of the programme. Local reactogenicity data compare favourably with other routine vaccinations and no specific increase in adverse reactions has been associated with their use in infant vaccination programmes. Self-limiting systemic reactions such as fever, myalgia, headaches and irritability have commonly been observed in prelicensure studies. Passive postlicensure safety monitoring of suspected adverse reactions has identified a large number of reports, generally of reactions deemed non-serious and reversible. An Expert Working Group has concluded the balance of benefits and risks to be overwhelmingly favourable. Further safety data are expected from large data-linkage studies. Present efficacy estimates, based on active surveillance of case numbers, vaccine failures and coverage rates, are approximately 90% for all age groups. A significant fall in the number of cases attributable to meningococcal group C infection has been observed in the age group of < 20 years. The annual number of fatalities from confirmed meningococcal C disease in the same population has fallen from 67 to 5 cases within a 2-year period. PMID- 12904122 TI - Neuroleptic malignant syndrome induced by atypical antipsychotics. AB - A review of the English literature confirms that neuroleptic malignant syndrome (NMS) occurs with both traditional and atypical antipsychotic medications. Published reports of NMS induced by the traditional antipsychotics have given the practitioner valuable information on the prevention and treatment of this adverse effect. Case reports have also been published concerning NMS and clozapine, risperidone, olanzapine and quetiapine. By evaluating the case reports of atypical antipsychotic-induced NMS, valuable information may be obtained concerning similarities or differences from that induced by the traditional antipsychotics. The case reports of NMS with atypical antipsychotics were evaluated for diagnosis, age/sex of patient, risk factors, antipsychotic doses and duration of use, symptoms of NMS, and clinical course. PMID- 12904123 TI - Efficacy and safety of midodrine in the treatment of dialysis-associated hypotension. AB - Dialysis-associated hypotension is a morbid problem in haemodialysis patients. Employment of midodrine at the start of dialysis has reduced the severity and frequency of hypotensive episodes in these patients. Through selective alpha(1) adrenergic receptor-binding, desglymidodrine, the active metabolite of midodrine, raises blood pressure by enhancing venous and arterial tone. This medication has been demonstrated to be effective and safe in the acute and chronic treatment of haemodialysis-associated hypotension in end stage renal disease patients. PMID- 12904124 TI - Risk-benefit analysis of platelet glycoprotein IIb/IIIa inhibitors in acute coronary syndromes. AB - Four intravenous glycoprotein IIb/IIIa receptor inhibitors (GPIs) (abciximab, eptifibatide, tirofiban and lamifiban) have been tested extensively over the last decade for their efficacy and safety in patients with acute coronary syndromes (ACS). GPIs are well-established adjunct agents for patients undergoing percutaneous coronary intervention, and considerable effort has gone into evaluating these agents in patients who are not scheduled to undergo coronary revascularisation. In the current article, six major randomised clinical trials conducted in the latter patient population are reviewed. Based on a recent meta analysis of these trials, GPIs reduced the incidence of death or myocardial infarction in patients not scheduled for early revascularisation, with the greatest reduction observed in patients at high risk of thrombotic complications. Major bleeding complications were more frequent in those receiving GPIs, however, the incidences of intracranial haemorrhage and stroke were similar in both treatment groups. Despite these risks, the benefits of GPI therapy in addition to conventional treatment, such as aspirin and heparin, should be considered for these high-risk patients. PMID- 12904126 TI - Safety issues surrounding the use of aromatase inhibitors in breast cancer. AB - Third generation aromatase inhibitors (AIs) are now established therapy in advanced oestrogen receptor (ER)-positive breast cancer. As the use of AIs expands to include adjuvant treatment of early breast cancer and breast cancer prevention, tolerability and effects on other organs such as bone will become as important as the antitumour properties of the drugs. In direct comparisons with tamoxifen, AIs have a better toxicity profile with fewer patients stopping therapy because of drug-related side effects. There is a lower incidence of thromboembolic events and vaginal bleeding compared with tamoxifen. Although published information about the side effects of AIs is scarce, it is likely that they will have adverse effects on bone and possibly also on lipid metabolism. Subprotocols of ongoing adjuvant trials are investigating these effects. It is likely that the choice of which third generation AI to use will be largely determined by its tolerability and safety profile, since it is likely that the currently available drugs have similar efficacy. PMID- 12904125 TI - Cardiotoxic effects of anthracycline-taxane combinations. AB - The association of doxorubicin (DOX) and paclitaxel (PTX) is very active in breast cancer. Unfortunately, PTX may potentiate the cardiotoxic effects of anthracyclines: it causes nonlinear disposition of DOX and its metabolites, leading to persistant of elevated plasma concentrations of the anthracyclines. However, this pharmacokinetic interference is not sufficient to explain the enhanced cardiotoxicity of the combination. Recent data suggest that PTX stimulates the conversion of DOX to cardiotoxic metabolites (namely doxorubicinol) inside cardiomyocytes. Docetaxel (DTX) does not have a major influence on DOX plasma concentration because it does not interfere with its elimination. Clinical data suggest that DTX may not enhance anthracycline cardiotoxicity, but patients seldom received a total anthracycline dose compatible with increased risk. Furthermore, there are experimental data indicating that DTX can also stimulate the metabolism of DOX to toxic species in human heart. PMID- 12904128 TI - 4th International Workshop on Adverse Drug Reactions and Lipodystrophy in HIV. San Diego, California, USA 22-25 September 2002. AB - Highlights are presented from this annual workshop, which was devoted to the investigation of adverse effects associated with antiretroviral therapy for HIV infection. Topics covered included the lipodystrophy syndrome, which encompasses body composition changes (subcutaneous fat wasting, visceral fat accumulation) and metabolic abnormalities (insulin resistance and dyslipidaemia). The relevance of HIV protease inhibitor-induced metabolic abnormalities to cardiovascular disease is discussed. Research in the areas of hyperlactataemia, abacavir hypersensitivity, and bone mineral density in the context of HIV infection is also briefly reviewed. PMID- 12904127 TI - Safety of low molecular weight heparins in the treatment of venous thromboembolism. AB - Low molecular weight heparins (LMWHs) are commonly employed as a substitute for unfractionated heparin (UFH) in the treatment of venous thromboembolic events. Despite their higher cost, the preferential use of LMWHs seemed justified initially as, based on the results of earlier meta-analyses, these compounds were deemed to be more effective and safer than UFH. Although, in this respect, their purported superiority over UFH could not be confirmed by subsequent large, randomised trials and updated meta-analyses, other peculiar features of LMWHs were highlighted, favouring their preferential utilisation in patients with venous thromboembolism. Among these, the possibility of once-daily administration on an out-patient basis, the lower incidence of Type II heparin-induced thrombocytopenia and the lower likelihood of osteoporosis after prolonged treatment periods, appear to be especially prominent. This review attempts to evaluate the available evidence focusing on the safety of LMWHs for the treatment of venous thromboembolism and the current therapeutic options and potential advantages of LMWHs, either in general or in selected patient populations. PMID- 12904129 TI - Zyban-- is there a cause for concern? AB - Bupropion hydrochloride is a monocyclic antidepressant which is used in a slow release formulation as a smoking cessation aid. Its side effects include insomnia and dry mouth with seizures occurring in 1 in 1000 patients. In overdose, it can cause tachycardia, conduction defects and convulsions. It is, however, effective in smoking cessation in approximately 20% of patients and may be more effective if combined with nicotine replacement therapy. The key question is whether the risks associated with the drug are justified by the obvious benefits to health from smoking cessation. PMID- 12904130 TI - Adverse effects of antiretroviral therapy: focus on orofacial effects. AB - The optimum anti-HIV drug has yet to be found. This paper will summarise some of the oral adverse effects associated with antiretroviral agents against HIV. The development of antiretroviral drugs for the treatment of HIV infection has been aimed at the inactivation of two HIV enzymes: reverse transcriptase and proteases. Erythema multiforme, ulcers and xerostomia are the main oral side effects associated with reverse transcriptase inhibitors. Parotid lipomatosis, taste disturbance, xerostomia and perioral paraesthesia are oral adverse effects, which are mainly related to protease inhibitor therapy. The search for new antiretrovirals with different active mechanisms and patterns of resistance constitutes a key question in HIV treatment. The use of new drugs and drug combinations will lead to the appearance of oral lesions, which will be difficult to identify and treat and which cannot be ignored by the practitioner. PMID- 12904132 TI - Adverse skin reactions to inhaled corticosteroids. AB - Inhaled corticosteroids are considered to be the therapy of choice in the treatment of asthma and allergic rhinitis. However, various cutaneous adverse reactions have been described and are probably present in approximately half of patients. Some of these reactions are related to the daily dosage of steroids or the duration of treatment, for example: thinning of the skin, easy bruising, acne or peri-oral dermatitis, mucocutaneous infection and, especially, candidiasis. Other cutaneous side effects are independent of the dose, such as allergic reactions and a few other rare side effects. Adverse effects could be prevented by monitoring the daily dosage and the family history of the patients, and by following advice provided on the use of inhaled corticosteroids. Specific treatments for infection, acne or allergy can cure these side effects. However, cutaneous side effects can become unpleasant for the patient and temporarily withdrawing therapy has proven to be extremely effective. PMID- 12904131 TI - Safety of fluoroquinolones in paediatrics. AB - In the last few years, there has been increasing pressure to use fluoroquinolones in paediatric patients, since these antibiotics offer the advantage of an oral treatment regimen on an out-patient basis. However, even although this class of antibiotics generally remains well-tolerated, the restriction of fluoroquinolone use in children on a compassionate basis, which derives from their potential to cause cartilage toxicity, limits the safety data in this population and suggests a cautious use. This review reports the data of the literature on the safety of fluoroquinolones in different districts, focusing on the side effects in children and drug interactions. Moreover, data available in the literature with regards to side effects in children are reported, with particular attention to their potential in arthropathy. PMID- 12904133 TI - Medication overuse headaches -- what is new? AB - Patients suffering from frequent migraine attacks or chronic tension-type headache are at risk of developing drug-induced headache (now called 'medication overuse headache' [MOH]) as described by the new classification of the International Headache Society. This headache entity is caused by frequent use of antiheadache compounds (non-opioid analgesics, ergot alkaloids, 5-HT agonists, combined preparations with caffeine or codeine) for more than 15 days/month. Most patients, however, use the medication daily. The standard therapy of MOH is withdrawal therapy (mostly under in-patient conditions) followed by prophylactic treatment of the primary headache. The relapse rate reaches up to 40% within 1 year after a previously successful withdrawal therapy. The pathophysiology of MOH remains to be determined and, consequently, therapy has been driven by experience rather than by scientific hypotheses. The identification of predictors for both medication overuse and relapse after successful withdrawal should improve prevention and therapy in the future. Several medical (e.g., class of overused drugs) and psychological (e.g., standard of performance) predictors were recently identified. Medication overuse in headache patients is based on multiple factors with physical, social and psychological impact. In view of the high prevalence and relapse rate, it is necessary to establish a structured post-treatment programme for patients after the acute withdrawal phase, which considers the predictors for relapse. PMID- 12904134 TI - Hepatotoxicity of chemotherapy. AB - The selection of a chemotherapeutic regimen for the oncology patient is based on a thorough assessment of potential hazards relating to the patient's clinical condition and the toxicities of chemotherapy. Liver function abnormalities are commonly seen in this patient population and deducing their aetiology may be difficult. Immunosuppression, paraneoplastic phenomena, infectious disease, metastases and polypharmacy may all confound the clinical picture. While criteria for standardising liver injury have been established, dose modifications often rely on empirical clinical judgement. Therefore, a comprehensive understanding of hepatotoxic manifestations for the most common chemotherapeutic agents is essential. This article reviews the hepatotoxicity of commonly utilised antineoplastic agents. PMID- 12904135 TI - Pharmacotherapy of gastro-oesophageal reflux disease in children: focus on safety. AB - Gastro-oesophageal reflux (GOR) disease is very common and, in the majority of infants, is physiological. However, untreated GOR disease in infants and children is associated with a decrease in quality of life for the child and their parents. It may also cause sometimes more severe complications, such as oesophagitis, and causing, in rare cases, failure to thrive, oesophageal stricture, apnoea and even death. Every therapeutic intervention (non-drug treatment, medical treatment and surgery) is associated with morbidity and even mortality. Moreover, efficacy data of many medications in children are non-existing, limited or disappointing. The safety profile of cisapride is comparable to that of other therapeutic interventions or to the risk of non-treatment. Therefore, the therapeutic approach of GOR disease in infants and children needs to be well-balanced, considering therapeutic efficacy and side effects. This review will focus on the side effects of the different therapeutic options. PMID- 12904136 TI - Controversy surrounding the safety of cerivastatin. AB - The noted myotoxicity and subsequent withdrawal of cerivastatin from the worldwide market in August 2001 has demonstrated that the safety of statins is not a class effect. The total rhabdomyolysis rate for cerivastatin was 16 - 80 times more frequent than with other statins without providing additional efficacy. Cerivastatin has a pharmacokinetic profile (high potency, bioavailability, lipophilicity and renal excretion) that is different from other statins, which may explain the high myotoxicity rate. The cerivastatin experience has also provided insights into high-risk populations (i.e., the elderly, women, those with renal impairment, co-administration of interacting drugs) that are more prone to statin-induced myopathy. Ultimately, the lessons learned from this experience may significantly improve the safety of statin use in the future. PMID- 12904137 TI - Safety of the new generation recombinant factor concentrates. AB - Haemophilia A and B are X-linked disorders resulting from deficiency of Factor VIII and IX, respectively. Clinical sequellae of Factor VIII or IX deficiency include spontaneous and traumatic haemorrhages into joints, soft tissues, and muscles. The cornerstone of therapy has been replacement of the deficient factor, historically with pooled-plasma derivatives. The unfortunate blood-borne infection transmission (such as HIV, hepatitis B and C viruses), inhibitor formation, immunosuppression, and, in certain cases, thrombosis by these products has spawned major advances and innovations in the manufacture of clotting products. Recombinant technology has virtually eliminated transmissible disease risk; yet, the presence of albumin in second and third generation recombinant products raises, at the least, theoretical risk of prions and parvovirus B19. Other non-infectious complications, including inhibitor formation, allergic reactions, and thrombosis, remain formidable concerns. Despite this, recombinant factors remain the most attractive treatment approach for haemophilia. Future improvement awaits the development of safe and effective gene transfer technology. PMID- 12904138 TI - Emerging safety issues regarding long-term usage of H(1)receptor antagonists. AB - The second generation histamine H(1)-receptor antagonists are important therapeutic tools in the treatment of atopic disease and may also have a place as an adjunct therapy for those patients whose allergic asthma coexists with allergic rhinitis. They are amongst the most widely prescribed and safest drugs in the world. However, as second generation H(1)-receptor antagonists are used to treat non-life threatening conditions, the risk of adverse effects is of vital importance. For many, the potential for sedation by some of the newer antihistamines still remains an issue, while there have recently been widespread concerns regarding the potential for cardiotoxicity and the impact of drug-drug interactions associated with some second generation H(1)-receptor antagonists. Consequently, progress with this class of drugs should involve not only increased efficacy but also improvements in their safety and specificity. Moreover, there is a trend towards using second generation H(1)-receptor antagonists as long-term therapy rather than confining their use to treating the short-term manifestations of allergic disease. To this end, a number of novel, potent and safe antihistamines have been developed which are either metabolites of active drugs or enantiomers. This review will examine some of the safety issues associated with established and newer second generation drugs particularly in relation to their long-term usage in adults and children. PMID- 12904139 TI - Inhaled steroids and the risk of adrenal suppression in children. AB - Corticosteroids are the mainstay of treatment of all asthma severity levels in adults and children. With their widespread use comes a responsibility to monitor, understand, and balance their efficacy and safety. Systemic adverse effects such as adrenal suppression have been clearly associated with the use of oral corticosteroids and to a lesser degree with the use of inhaled corticosteroids (ICS). In clinical trials, adrenal suppression is more evident when ICS are used in long-term therapy and at higher doses. However, monitoring adrenal suppression during short-term therapy and at lower doses is still of value in order to ascertain the lower limit of an inhaled corticosteroid's safety profile. Significant adrenal suppression at conventional ICS doses appears to be rare in clinical practice. When evaluating the effect of ICS on the hypothalamo-pituitary adrenal-axis (HPA-axis), one must consider sources of variability both within and among trials including test sensitivity, systemic bioavailability, degree of airway obstruction, and delivery devices. All of these factors have the potential to effect the level of adrenal suppression detected and must be considered when interpreting HPA-axis test results in research or practice. This review will discuss adrenal suppression found with common ICS. PMID- 12904140 TI - Treatment and prevention of aspirin-induced gastroduodenal ulcers and gastrointestinal bleeding. AB - Aspirin use is associated with gastroduodenal mucosal damage and increased risk of upper gastrointestinal (GI) bleeding. Many aspirin users should receive prophylactic treatment since they often have several risk factors for upper GI complications. The best therapeutic approach for reducing GI toxicity in low-dose aspirin users is still ill-defined as only a few studies have focused on this problem. Omeprazole appears to be very effective in reducing both acute gastroduodenal mucosal damage and upper GI bleeding in the high-risk patient taking low-dose aspirin, but data with other anti-ulcer agents are lacking (misoprostol) or inconsistent (ranitidine) at present. The role of Helicobacter pylori is controversial in NSAID users, but there is now wide agreement that H. pylori infection increases mucosal damage and the risk of upper GI bleeding in low-dose aspirin users. PMID- 12904141 TI - Breast cancer chemoprevention: risk-benefit effects of the antioestrogen tamoxifen. AB - The anti-oestrogen tamoxifen, which is widely used as adjuvant therapy for breast cancer, is undergoing evaluation as a chemopreventive agent in women at increased risk of developing this disease. Recent results from the National Surgical Adjuvant Breast and Bowel Project (NSABP) P-1 prevention trial show a 49% reduction in breast cancer incidence in healthy, high-risk women. However, tamoxifen treatment has the serious side effect of increasing the incidence of endometrial cancer in women and long-term administration of tamoxifen causes hepatic tumours in rats. These liver tumours are induced via a genotoxic mechanism, but the mechanisms responsible for endometrial cancer in women are not yet known and are a focus of much debate. This review describes the findings from the chemoprevention trials and problems associated with the use of tamoxifen in this setting. The mechanism of carcinogenesis in rat liver is explained in detail and compared to the situation in humans, with a view to assessing the risks associated with tamoxifen therapy and predicting whether other anti-oestrogens might be safer alternatives. PMID- 12904142 TI - A safety look at currently available statins. AB - In this mini-review, the evidence for safety and efficacy of currently available statins is discussed. Large-scale, long-term clinical studies have documented an outstanding efficacy and safety profile for statin monotherapy when used at pharmacological doses. Non-life-threatening side effects may occur in up to 15% of patients receiving one statin. Sporadic reports show possible adverse effects of statins on nervous system function including mood alterations. More serious side effects may also occur but at much lower rates. Significant elevations in the activity of serum aminotransferase and creatine kinase alone or in combination with muscle pain in statin-treated patients should be taken seriously; under these conditions, monitoring the statin dose or its discontinuation must be considered. Unlike monotherapy, combination therapy is more problematic. Particularly, combination of the statins with gemfibrozil results in higher rates of drug toxicity. Co-administration of statins with other drugs, especially those which may interfere with the cytochrome P450 system, should be considered carefully. Special attention must be paid to the tolerability of the statins in elderly and transplant patients. The safety of statins in children and adolescents has not yet been well-documented, thus, statin therapy is not routinely recommended in this group of hyperlipidaemic subjects. Future clinical studies and surveillance information will warrant long term safety of each member of this class of lipid-lowering agents. PMID- 12904143 TI - Toxicogenomics: an opportunity to optimise drug development and safety evaluation. AB - 'Toxicogenomics', the use of complex populations of mRNA to understand toxicity, is a relatively new field which combines the wealth of gene sequence information with advances in miniaturisation technology. In a parallel evolutionary path that the broader field of toxicology has encountered, toxicogenomics is steadily changing from descriptive to mechanistic research and will ultimately progress to a predictive science. This review focuses on the application of microarray research tools to toxicology studies in preclinical development. In particular, the 'testing funnel' approach to candidate selection in drug development is used to discuss the strategic implementation of toxicogenomics to help develop drugs that harbour less toxicity. Examples of where toxicogenomics has increased mechanistic and descriptive understanding of cellular toxicity are discussed, as are early efforts to develop molecular biomarkers that are predictive of longer term toxicity. PMID- 12904144 TI - Anticoagulant-related skin reactions. AB - Cutaneous reactions have been reported during anticoagulant therapy with coumarin derivatives, unfractionated and low molecular weight heparins, heparinoids, danaparoid and hirudins. Anticoagulant-induced skin reactions vary from local allergic manifestations to skin necrosis. In patients with allergic reactions, diagnosis and crossreactions between anticoagulants can be confirmed by intracutaneous testing. Coumarin- and heparin-induced skin necrosis are rare, but are important side effects due to their high morbidity and occasional mortality. Cutaneous tests are contraindicated in these patients. In the future, anticoagulant strategies may include direct synthetic thrombin inhibitors (argatroban and melagatran/ximelagatran) and the Factor Xa inhibitor, pentasaccharide (fondaparinux). PMID- 12904145 TI - MMR vaccination and autism: is there a link? AB - In 1998, a report was published describing 12 patients with inflammatory bowel conditions and regressive developmental disorders consisting primarily of autism. The authors hypothesised that MMR vaccine may have been responsible for the bowel dysfunction which subsequently resulted in the neurodevelopmental disorders. The suggestion that measles vaccine may cause autism through a persistent bowel infection generated much interest since it provided a possible biological mechanism for a causal association. Epidemiological studies, however, have not found an association between MMR vaccination and autism. Autism has a strong genetic component and its associated neurological defects probably occur during embryonic development. It seems unlikely that a vaccination that is given after birth could cause autism. In a minority of cases, autism may have onset after 1 year of age (regressive autism) but the one epidemiological study that included such cases did not find an association with MMR vaccination. Currently, the weight of the available epidemiological and related evidence does not support a causal link between MMR vaccine and autism. PMID- 12904146 TI - Cardiotoxicity of macrolides, ketolides and fluoroquinolones that prolong the QTc interval. AB - Macrolides, ketolides and fluoroquinolones as well as other classes of antimicrobial agents have been associated with prolongation of cardiac repolarisation. This effect is most notable with erythromycin, clarithromycin, gatifloxacin, moxifloxacin, levofloxacin and telithromycin. All of these agents produce a blockage of the HERG channel dependent potassium current in myocyte membranes resulting in a prolonged QTc interval which may give rise to polymorphic ventricular tachycardia, Torsades de Pointes or ventricular fibrillation. The risk of malignant arrhythmias is increased by concomitant usage with Type Ia or III anti-arrhythmic agents or with other drugs that prolong the QTc interval or have competitive metabolic routes. Electrolyte disturbances or underlying cardiac disease also increase the risk of ventricular arrhythmias. The best clinical outcome indicator is the incidence of the associated arrhythmias. The rough rank order of risk with these agents, albeit with limited and incomplete data, is in decreasing order; erythromycin, clarithromycin, gatifloxacin, levofloxacin and moxifloxacin. Telithromycin outcomes for associated arrhythmia are yet to be determined. The essential point is that the overall risk of ventricular arrhythmias is very small with these agents but can be reduced further by avoiding their usage for patients with other multiple risk factors for Torsades de Pointes. PMID- 12904147 TI - Management of haemorrhagic complications associated with oral anticoagulant treatment. AB - The frequency of anticoagulant-related bleeding has been reported to range 1-7% per year, depending on the indication for anticoagulant therapy and the classification of bleeding. Although most bleeding is not life threatening, it does cause short-term morbidity and inconvenience to patients, as well as possibly diminishing their quality of life to some degree. Assessing the risk of anticoagulant-related bleeding is integral to optimising the management of anticoagulant therapy. Furthermore, early recognition and treatment of anticoagulant-related bleeding can reduce the associated morbidity. This article reviews the definitions of major and minor bleeding, the assessment of risk and preventive strategies and the management of anticoagulant-related bleeding. PMID- 12904148 TI - Risk-benefit assessment of angiotensin II receptor antagonists. AB - Inhibiting the renin-angiotensin-aldosterone system through the use of angiotensin-converting enzyme (ACE) inhibitors has proven very useful in the treatment of hypertension, congestive heart failure (CHF) and progressive renal failure. More recently, agents that directly block the angiotensin II Type 1 (AT(1)) receptor--angiotensin II receptor antagonists (AIIRAs)--have been developed. These agents are thought to have a more specific mechanism of action since they do not affect other hormone systems as do the ACE inhibitors. Whether such specificity results in a different efficacy profile is still being determined. However, these drugs are extremely well-tolerated and very safe. AIIRAs are effective in the reduction of both systolic and diastolic blood pressure and compare favourably to other classes of agents. Recent results indicate that at least one AIIRA has a favourable effect on stroke in hypertensive patients with left ventricular hypertrophy. Additional studies with other members of the class will provide further information on similar outcomes. In CHF patients, ACE inhibitors remain the drug of choice and AIIRAs are best utilised in patients who cannot tolerate an ACE inhibitor or in those receiving an ACE inhibitor who cannot tolerate a beta-blocker and need additional therapy. AIIRAs are effective in slowing the progression of renal failure in patients with Type II diabetes and may be effective in other proteinuric conditions. Whether they are more or less effective than ACE inhibitors is unknown. Overall, AIIRAs represent an important addition to the armamentarium of cardiovascular therapies with an excellent safety record and an emerging profile of utility in multiple cardiovascular conditions. PMID- 12904149 TI - IVIG in autoimmunity and cancer--efficacy versus safety. AB - Intravenous immunoglobulin (IVIG) is an accepted treatment for certain immunodeficiency states, but also for some autoimmune diseases. In other autoimmune conditions (i.e., systemic lupus erythematosus), it is still empirical although very efficacious. Based mainly on animal studies, IVIG has been shown to exert different antitumour mechanisms that result in metastases suppression. This effective therapy is associated with frequent occurrence of either immediate, delayed or late adverse effects, most of which are mild and transient. Efforts are taken in order to minimise these adverse effects both by pharmaceutical companies that attempt to decrease the risk of infectious agents transmission, and by physicians who monitor closely patients and choose the appropriate mode of administration of IVIG with respect to dose, duration of treatment and preparations used. Overall, IVIG is used in severe medical conditions and it is quite safe. Future research would help to further minimise its associated risks. PMID- 12904150 TI - The rational use of potentially hepatotoxic medications in patients with underlying liver disease. AB - Given the fact that as many as 9% of all adverse drug reactions involve toxic effects on the liver and with upwards of 50% of all cases of fulminant hepatic failure being ascribed to acetaminophen and other agents, the safe use of medications takes on an even greater importance whenever the prescription of potentially hepatotoxic drugs to patients with underlying liver disease is considered. In general, it is thought that most drugs can be safely administered in the setting of liver disease without an increased risk of hepatotoxicity, although the evidence on which this statement is based often relies more on clinical judgement than on clinical studies. Several drugs appear to have an increased risk of hepatotoxicity in patients with underlying liver disease based on either clinical reports or extrapolated pharmacological data. These agents, including methotrexate, niacin and the antiretroviral and antituberculosis drugs, carry warnings about their use in patients with a variety of liver conditions. The data supporting the hepatotoxic risk of scores of additional drugs, such as the 3-Hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase inhibitors ("statins"), the newer thiazolidinediones (rosiglitazone, pioglitazone), and tamoxifen, among others, in patients with liver disease are generally lacking by evidence-based studies. However, clinical and biochemical monitoring is routinely recommended or required, often to make up for the lack of information on the true risk of clinically significant liver toxicity of these agents in individuals both with and without underlying liver disease. This article will review what is and what is not known about prescribing in the setting of acute and chronic liver disease and offers recommendations to help promote the safe and rational use of potentially hepatotoxic medications in these patients. PMID- 12904151 TI - Treatment of drug-induced thrombocytopenia. AB - Estimates on the incidence of drug-induced thrombocytopenia range 5-40% in patients receiving heparin to < 1% with other causative agents. Systematically assessing drug-induced thrombocytopenia through a series of steps, each step providing additional evidence that the suspected agent is the true cause of thrombocytopenia, is the best way to identify the causative agent. Databases exist to aid in identification of the causative agent. Knowing which medications may be causative agents as well as which are not known to cause drug-induced thrombocytopenia, the aetiologies of drug-induced thrombocytopenia, signs and symptoms of thrombocytopenia and strategies to treat thrombocytopenia associated with specific agents will provide the clinician with the necessary skills to make proper medical decisions. PMID- 12904152 TI - Short- and long-term safety of glaucoma drugs. AB - Glaucoma, a leading cause of blindness worldwide, is a chronic neurodegenerative disorder. Patients with glaucoma may require long-term administration of intraocular pressure (IOP)-lowering medications. These medications belong to several classes of molecules including beta-adrenergic blockers, cholinergic agents, alpha-adrenergic agonists, carbonic anhydrase inhibitors and ocular hypotensive lipids. Most adverse effects associated with IOP-lowering medications are mild and ocular in nature; however, several of them are associated with systemic risks as well as serious ocular effects, especially following chronic use. The following review discusses the acute and long-term effects of commonly used IOP-lowering medications. PMID- 12904153 TI - 1st International Symposium on the Evaluation of Safety of Human Vaccines. 22-23 May 2002, Istituto Superiore di Sanita, Rome, Italy. AB - The first international meeting to focus exclusively on vaccine safety, brought together vaccine surveillance specialists, trialists, pharmaceutical companies, regulators, researchers and academics to examine and discuss methodological problems in the assessment of the safety of human vaccines. The 1st International Symposium on the Evaluation of Safety of Human Vaccines took place in Rome, Italy on 22 - 23 May 2002. The Istituto Superiore di Sanita (ISS), Rome, Italy, sponsored the symposium. The symposium was opened by D Greco (Head of Epidemiology and Biostatistics, ISS and a well-known vaccine trialist) who spoke of the need for the gathering and sharing of information regarding adverse effects of vaccines, as single studies "are not enough" and "have their limitations". Vaccines have come along way since their first use, however, as there is public concern over real or perceived adverse events, collaboration is of great importance as uncertainty about the effectiveness and safety may affect coverage of immunisation programmes. PMID- 12904154 TI - Return of alosetron. PMID- 12904155 TI - Risk-benefit of HMG-CoA reductase inhibitors in the treatment of HIV protease inhibitor-related hyperlipidaemia. AB - HIV protease inhibitors decrease mortality and improve quality of life in patients with HIV infection. However, these drugs have been associated with serum lipid elevations, which may pose an increased risk of cardiovascular disease and pancreatitis. Treatment of protease inhibitor-related hyperlipidaemia (PIH) is complicated by drug interactions, which significantly increase concentrations of most 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase inhibitors (statins). Although pravastatin and atorvastatin effectively lower cholesterol and triglyceride concentrations in HIV-infected patients, a significant number of patients did not achieve their National Cholesterol Education Program low density lipoprotein concentration goals. Nonetheless, due to the increased risk of rhabdomyolysis with elevated statin concentrations, atorvastatin should be considered a second-line agent. The limited available PIH data supports the fact that pravastatin and atorvastatin are well-tolerated in HIV-infected individuals. More data are needed on the appropriate starting doses, maximum safe doses, role of combination statin-fibrate therapy, documentation of coronary heart disease benefit and incidence of myotoxicity and hepatotoxicity. Pravastatin has an acceptable risk-benefit ratio in PIH, while theoretical toxicity concerns exist with atorvastatin. PMID- 12904156 TI - Safety evaluation of the drugs available to prevent malaria. AB - All drugs used for malaria prophylaxis have common adverse effects, in addition to rare and/or severe adverse effects. For many of the drugs in current use, the common adverse effects include neuropsychiatric harms. This property makes these drugs unpopular with tourists and business travellers, most of whom will be well at the start of chemoprophylaxis. Drugs available to prevent malaria have not been rigorously researched in terms of the phenomenology of their unwanted effects. Consequently, prescribers are not well placed to give useful information to travellers on the incidence, natural history and avoidability of the harms they may experience from malaria chemoprophylaxis. There is some evidence that the adverse effects of mefloquine may be a post-hepatic syndrome caused by drug induced liver damage with, in some users, symptomatic thyroid disturbance. However, confusion in the interpretation of the scientific evidence has led to conflicting messages regarding the safety of mefloquine and other antimalaria drugs, and to incorrect self-therapy by individual travellers, sometimes with fatal outcomes. In this review, the existing knowledge base for the safety of drugs currently used to prevent malaria is described along with present designs for future studies that would allow a rigorous safety assessment of candidate chemoprophylactic agents and of new drugs introduced to prevent malaria. There is an urgent need for internationally-agreed, evidence-based malaria prevention guidelines. These guidelines should be explicitly linked to the best available research evidence (normally systematic reviews of trials and individual randomised trials) and should highlight gaps in the knowledge base as priority areas for research. PMID- 12904157 TI - Long-term safety of proton pump inhibitors: risks of gastric neoplasia and infections. AB - After Helicobacter pylori eradication was introduced and largely eliminated the need for maintenance therapy for peptic ulcer disease, gastroesophageal reflux disease (GERD) became the main indication for prolonged gastric acid inhibition. The drug effect on GERD depends on the degree of acid inhibition, thus the efficacious proton pump inhibitors are preferred. The proton pump inhibitors have few immediate side effects, the main concern being the profound hypoacidity and hypergastrinaemia they induce. In short-term, hypergastrinaemia causes rebound hyperacidity, possibly worsening GERD and reducing the efficacy of histamine H(2) blockers. In the long-term, hypergastrinaemia causes enterochromaffin-like cell hyperplasia and carcinoids. Since enterochromaffin-like cells may be important in gastric carcinogenesis, iatrogenic hypergastrinaemia may predispose to carcinoma. Gastric hypoacidity also increases gut bacterial infections, and the barrier function of acid against viral and prion infections requires further assessment. PMID- 12904158 TI - Safety of the newer antihypertensive agents in children. AB - Hypertension is relatively uncommon in children and few children receive antihypertensive medications. This article reviews the safety of calcium channel blockers, angiotensin-converting enzyme inhibitors and angiotensin II receptor antagonists in children with hypertension. While the newer antihypertensive agents appear to be well-tolerated by children, further studies are needed to determine the safety profile across the developmental continuum, with chronic dosing and in children with complex hypertension. PMID- 12904159 TI - Cardiovascular hazard of selective COX-2 inhibitors: myth or reality? AB - Since 1998, two selective inhibitors of COX-2 have been approved in many countries for the treatment of rheumatoid arthritis, osteoarthritis and acute pain. These new drugs have a significantly reduced gastrointestinal toxicity when compared with non-selective COX inhibitors. However, the results of two large clinical trials conducted in patients with osteoarthritis and rheumatoid arthritis have recently raised some concerns regarding the cardiovascular safety of these new drugs. The purpose of this paper is to review the potential mechanisms whereby selective COX-2 inhibitors could increase the cardiovascular risk of patients and to analyse the data indicating that this clinical risk indeed exists. The authors' analysis shows that even though there are pathophysiological mechanisms which could explain why selective COX-2 inhibition might increase the cardiovascular risk in patients, the actual level of evidence demonstrating that the risk is indeed increased is weak. Because of the importance of the issue, additional studies must be conducted with this class of agents. Meanwhile, it is crucial to emphasise that neither selective COX-2 inhibitors nor conventional NSAIDs replace aspirin in patients with a high cardiovascular risk. PMID- 12904160 TI - COX-2 selective inhibitors: analysis of the renal effects. AB - COX-2 selective inhibitors provide analgesia and blunt inflammation while also sparing the gastrointestinal tract from classic NSAID toxicity. Therapeutic effects are thought to result from inhibition of the inflammatory COX-2 isoform. Organ sparing is considered the result of preservation of homeostatic COX-1 enzyme function. Similar roles of the COX isoforms in the kidney would reduce NSAID-associated nephrotoxicity. However, human kidney tissue expresses COX-2 enzyme, suggesting a role for this isoform in maintenance of physiological renal processes. Available clinical data on the renal effects of COX-2 selective inhibitors in humans also demonstrate nephrotoxic potential. PMID- 12904161 TI - Drug-induced myopathies. AB - Drug-induced muscle disorders represent a broad clinical spectrum, from asymptomatic elevated serum creatine kinase levels to life-threatening myopathies. An increasing number of drugs have been suspected or identified as myotoxic. It would be unrealistic to attempt to list them. In clinical practice, an iatrogenic origin must be discussed in any patient presenting with muscle symptoms in as much as drug-induced myopathies are usually reversible after discontinuation of the offending compound. The clinical and histopathological features depend on the causative agent and individual susceptibility to a given compound. Apart from isolated myalgias, drug-induced myopathies can be divided into five major categories: necrotising myopathies mainly due to lipid-lowering drugs; inflammatory myopathies, including polymyositis (especially associated with thiol compounds) and maphrophagic myofasciitis; mitochondrial myopathies, mainly due to antiretroviral nucleoside analogues; corticosteroid myopathy; and various forms of painless neuromyopathies. In some cases (e.g., statin-induced myopathies), risk factors have been clearly identified and preventive measures have been recommended. PMID- 12904162 TI - The gastrointestinal tolerability and safety of oral bisphosphonates. AB - Oral bisphosphonates are effective for osteoporosis and other hyperresorptive bone disorders. Although well-tolerated in efficacy trials, some oral aminobisphosphonates have been associated with upper gastrointestinal intolerance and injury in postmarketing experience. Clinical trials often underestimate the rate of adverse events in clinical practice, and ethics prohibit direct evaluation of toxicity in high-risk patients. Accordingly, animal models and endoscopy studies of oral bisphosphonates provide valuable insight. It is unclear whether variation in ulcerogenic potential reflects differences in dosing, formulation or chemical structure. Furthermore, the clinical relevance of endoscopic lesions is uncertain. Ongoing postmarketing review will determine whether differences in endoscopic damage predict tolerability and safety in clinical practice. However, physicians and patients should consider risk factors for oesophageal injury when initiating therapy. PMID- 12904163 TI - Safety of adeno-associated virus gene therapy vectors: a current evaluation. AB - An increasing number of strategies for molecular treatment of disease rely on the adeno-associated virus (AAV) as a therapeutic gene delivery vector. One of the most attractive features of this viral DNA vector is the perceived safety of AAV gene delivery. Recent applications in human clinical trials support the safety record established in preclinical trials, with evidence of gene transfer in the absence of cellular immune responses or tissue disturbance. Nevertheless, many aspects of the biology of the wild type AAV and its derivatives are still being explored. While the therapeutic potential of novel recombinant AAV therapeutics appears promising, recent insights suggest aspects of their pharmacokinetics, biodistribution and toxicity that require consideration to achieve the safest application of these molecular medicines. PMID- 12904164 TI - Pharmacotherapy of osteoporosis in postmenopausal women: focus on safety. AB - The therapy of osteoporosis has made enormous strides in the last decade. There is now a range of interventions, each with its pros and cons. Calcium and vitamin D supplementation remain the foundation and have few safety issues. Bisphosphonates are widely used, though gastrointestinal tolerance is a problem with some oral preparations. Intravenous administration may circumvent this, although this introduces the smaller problem of acute phase reactions. The side effect profile of hormone replacement therapy (HRT) is still being delineated after 40 years of use, with substantial new information expected in the next few years. This will clarify its place in the medical management of the menopause. Raloxifene appears to have a superior safety profile to HRT, though its efficacy on bone may be less. While none of these options is suitable for everyone, the range of available therapies does mean that most patients can find an intervention that is effective and acceptable. PMID- 12904165 TI - The CCN family: a new stimulus package. AB - The CCN family comprises cysteine-rich 61 (CYR61/CCN1), connective tIssue growth factor (CTGF/CCN2), nephroblastoma overexpressed (NOV/CCN3), and Wnt-induced secreted proteins-1 (WISP-1/CCN4), -2 (WISP-2/CCN5) and -3 (WISP-3/CCN6). These proteins stimulate mitosis, adhesion, apoptosis, extracellular matrix production, growth arrest and migration of multiple cell types. Many of these activities probably occur through the ability of CCN proteins to bind and activate cell surface integrins. Accumulating evidence supports a role for these factors in endocrine pathways and endocrine-related processes. To illustrate the broad role played by the CCN family in basic and clinical endocrinology, this Article highlights the relationship between CCN proteins and hormone action, skeletal growth, placental angiogenesis, IGF-binding proteins and diabetes-induced fibrosis. PMID- 12904166 TI - IGF-binding protein-4: biochemical characteristics and functional consequences. AB - IGFs have multiple functions regarding cellular growth, survival and differentiation under different physiological and pathological conditions. IGF effects are modulated systemically and locally by six high-affinity IGF-binding proteins (IGFBP-1 to -6). Despite their structural similarity, each IGFBP has unique properties and exhibits specific functions. IGFBP-4, the smallest IGFBP, exists in both non-glycosylated and N-glycosylated forms in all biological fluids. It is expressed by a wide range of cell types and tIssues, and its expression is regulated by different mechanisms in a cell type-specific manner. IGFBP-4 binds IGF-I and IGF-II with similar affinities and inhibits their actions under almost all in vitro and in vivo conditions. In this review, we summarize the available data regarding the following aspects of IGFBP-4: genomic organization, protein structure-function relationship, expression and its regulation, as well as IGF-dependent and -independent actions. The biological significance of IGFBP-4 for reproductive physiology, bone formation, renal pathophysiology and cancer is discussed. PMID- 12904167 TI - Experiments using microarray technology: limitations and standard operating procedures. AB - Microarrays are a powerful method for the global analysis of gene or protein content and expression, opening up new horizons in molecular and physiological systems. This review focuses on the critical aspects of acquiring meaningful data for analysis following fluorescence-based target hybridisation to arrays. Although microarray technology is adaptable to the analysis of a range of biomolecules (DNA, RNA, protein, carbohydrates and lipids), the scheme presented here is applicable primarily to customised DNA arrays fabricated using long oligomer or cDNA probes. Rather than provide a comprehensive review of microarray technology and analysis techniques, both of which are large and complex areas, the aim of this paper is to provide a restricted overview, highlighting salient features to provide initial guidance in terms of pitfalls in planning and executing array projects. We outline standard operating procedures, which help streamline the analysis of microarray data resulting from a diversity of array formats and biological systems. We hope that this overview will provide practical initial guidance for those embarking on microarray studies. PMID- 12904168 TI - The role of apoptosis-inducing receptors of the tumor necrosis factor family in thyroid cancer. AB - The tumor necrosis factor (TNF) family comprises several ligands, such as the prototype TNF-alpha, the Fas ligand (FasL) and TNF-related apoptosis-inducing ligand (TRAIL/Apo2L), which trigger apoptosis in susceptible cells by activating respective cell-surface receptors. The study of these cell death pathways has attracted significant attention in several fields, including that of thyroid cancer, because they participate in immune system function, as an arm of cell mediated cytotoxicity, and because recombinant ligands are available for pharmacological use. TNF-alpha is a pluripotent cytokine that induces both pro apoptotic and anti-apoptotic effects on thyroid carcinoma cells. FasL triggers apoptosis in other tumor types, but thyroid carcinoma cells are resistant to this effect. On the other hand, TRAIL potently and selectively kills thyroid carcinoma cells. Consequently, TRAIL is the only member of the family with significant anticancer activity and an acceptable toxicity profile to be used as a novel therapy for thyroid cancer. The caspase inhibitor FLIP plays a significant role in negatively regulating receptor-induced apoptosis. Thelper 1-type cytokines, such as interferon-gamma, TNF-alpha and interleukin-1beta increase the sensitivity of both normal and neoplastic thyrocytes to FasL and TRAIL. On the other hand, IGF-I and other growth/survival factors produced in the local tumor microenvironment activate the phosphatidylinositol 3-kinase/Akt kinase pathway and exert an anti-apoptotic effect by upregulating several apoptosis inhibitors, including FLIP. Pharmacological modulation of apoptosis induced by FasL and TRAIL/Apo2L holds promise of therapeutic applications in human malignancies. PMID- 12904169 TI - Differential distribution and expression of leptin and the functional leptin receptor in major salivary glands of humans. AB - Leptin plays a central role in the regulation of food intake and energy expenditure in rodents. However, it has become clear that this hormone has more than only a satiety-inducing function, and that there are other sources of leptin, such as the central nervous system, placenta and the gastrointestinal tract in addition to adipose tIssue. Knowing about the important role of the salivary glands in food intake and digestion, it was the objective of the present study to investigate how leptin and its receptor are expressed and distributed in the major salivary glands of humans. We found leptin distributed throughout the major salivary glands with obvious intracellular concentrations in granula. In contrast, immunostaining for the leptin receptor was found exclusively in the membranes of the glandular cells. A high density of the leptin receptor was localised in the epithelia of the duct lumen. PCR analysis proved the autonomous expression of leptin by the salivary glands independently from adipocytes. Accordingly the long receptor isoform was expressed by any examined tIssue. In the light of recent findings of leptin influencing the growth of rodent salivary glands, the presence and distribution of leptin and its receptor suggests an autocrine role of salivary leptin within the glands. PMID- 12904170 TI - Lactation modulates diurnal expression profiles of specific leptin receptor isoforms in the rat hypothalamus. AB - We investigated the effects of lactation on diurnal changes in serum leptin and hypothalamic expression of the leptin receptor isoforms, Ob-Ra, -Rb, -Rc, -Re and -Rf in rats. In non-lactating rats, serum leptin concentration was increased at night while hypothalamic mRNA levels of Ob-Rb, -Rc and -Re decreased; by contrast, expression of Ob-Ra and Ob-Rf was unchanged at night. There were significant negative correlations between serum leptin and mRNA expression of Ob Rb (P<0.001) and Ob-Re (P<0.05), which were independent of time of day. In lactating rats, the nocturnal rise in serum leptin was attenuated. Daytime hypothalamic Ob-Rb mRNA levels were significantly lower than in non-lactating controls, and the normal nocturnal decreases in expression of Ob-Rb, -Rc and -Re were lost. The relationship between serum leptin and Ob-Re expression was not changed by lactation. Lactation had no effect on the expression of Ob-Ra mRNA in the hypothalamus. Decreased daytime Ob-Rb expression could lead to reduced hypothalamic sensitivity to leptin, and thus contribute to increased daytime appetite in lactating rats. Moreover, maintaining high levels of Ob-Re expression could, by increasing hypothalamic leptin-binding protein concentration and reducing local leptin bioavailability, further accentuate hyperphagia. Thus, selective changes in expression of specific isoforms of the leptin receptor may contribute to the hyperphagia of lactation in rats. PMID- 12904171 TI - Impaired mammary function and parathyroid hormone-related protein during lactation in growth-restricted spontaneously hypertensive rats. AB - Evidence implicates pivotal roles for parathyroid hormone-related protein (PTHrP) during lactation, including stimulation of mammary and pup growth. As spontaneously hypertensive rat (SHR) pups are growth restricted compared with the control Wistar Kyoto (WKY), we examined the relative roles of pup suckling and maternal lactational environment on pup growth, mammary PTHrP, and milk PTHrP and calcium concentrations. SHR pups were lighter compared with the control from 6 days. SHR mammary PTHrP content and milk PTHrP were lower but maternal plasma PTHrP was raised compared with WKY. SHR mammary morphological development was also impaired compared with control. Cross fostering growth-restricted pups onto WKY mothers increased pup weight in association with normal mammary function and higher milk PTHrP and calcium. Control pups suckling on an SHR mother had reduced body weight. Both cross fostering groups were associated with increased maternal and milk PTHrP concentrations, indicating the importance of suckling, together with a functional mammary gland. The results suggested that impaired SHR mammary function and milk PTHrP are associated with a reduced SHR postnatal growth. Our data also indicated that milk and mammary PTHrP are regulated by different mechanisms but that they are influenced by the maternal lactational environment and the suckling pup. PMID- 12904172 TI - Parathyroid hormone (PTH) down-regulates PTH/PTH-related protein receptor gene expression in UMR-106 osteoblast-like cells via a 3',5'-cyclic adenosine monophosphate-dependent, protein kinase A-independent pathway. AB - Parathyroid hormone (PTH) regulates osteoblast function via a G protein-linked PTH/PTH-related protein (PTHrP) receptor. We have studied the mechanisms of PTH/PTHrP receptor gene repression by PTH in UMR-106 osteoblast-like cells. Inhibition of PTH/PTHrP receptor mRNA expression by rat (r) PTH(1-34) and Insulin like growth factor-I (IGF-I) at 10(-7)M was significant at 1 h and 3 h, and maximal at 2 h and 6 h. A maximal decrease in receptor mRNA abundance by rPTH(1 34) and IGF-I was maintained for 24 h. Inhibition of receptor gene expression by rPTH(1-34) was mimicked in UMR-106 cells by the addition of forskolin (an adenylyl cyclase activator), or 8-(4-chlorophenylthio)-adenine 3',5'-cyclic monophosphate (8-pCPTcAMP; a cAMP analogue). Although H89, a selective protein kinase A (PKA) inhibitor, completely inhibited PKA activity stimulated by rPTH(1 34), forskolin or 8-pCPTcAMP, suppression of PTH/PTHrP receptor mRNA synthesis induced by these substances in UMR-106 cells was not affected by H89. In primary osteoblast cultures, rPTH(1-34) inhibited synthesis of PTH/PTHrP receptor mRNA irrespective of H89. The down-regulation effect of rPTH(1-34) was also unaltered by PD98059 (an extracellularly regulated kinase 1/2 mitogen-activated protein kinase pathway inhibitor). Pretreatment with cycloheximide, a protein synthesis inhibitor, did not alter the inhibition of PTH/PTHrP receptor mRNA expression by rPTH(1-34), indicating that receptor mRNA suppression does not require new protein synthesis. Transcriptional activation of PTH/PTHrP receptor gene promoter (U3P or U4P)-luciferase constructs was decreased by rPTH(1-34), forskolin and 8 pCPTcAMP irrespective of H89. Thus, PTH transcriptionally down-regulates PTH/PTHrP receptor gene expression in osteoblast-like cells via a cAMP-dependent, PKA-independent pathway. PMID- 12904173 TI - Regulation of mammary tight junctions through parathyroid hormone-related peptide induced activation of apical calcium channels. AB - Tight junctions (TJs) play an essential role in cell-cell contact between mammary epithelial cells and, as such, play a critical role in cell function. Moreover, calcium (Ca) plays a crucial role in the formation and maintenance of mammary TJs. Given that parathyroid hormone-related peptide (PTHrP) is involved in cellular Ca homeostasis, we postulated a role for PTHrP in the regulation of mammary TJs. The effect of PTHrP(1-34) on TJs was studied in the mouse mammary cell line COMMA-1D by measuring transepithelial electrical resistance across cell monolayers and measuring the expression of TJ proteins. PTHrP stimulated TJ formation but only under conditions where extracellular Ca was limiting. This effect of PTHrP appeared to be indirect and mediated via increased intracellular availability of Ca as a result of increased Ca-channel activity in the apical membrane. The changes in TJs were associated with altered expression of the TJ protein occludin, but expression of the TJ protein claudin-1 was not affected. The effects of PTHrP on mammary TJs are independent of prolactin. In conclusion, PTHrP enhances mammary TJ formation when extracellular Ca is limiting by maintaining intracellular Ca supplies. PMID- 12904174 TI - Prolactin regulation of Bcl-2 family members: increased expression of bcl-xL but not mcl-1 or bad in Nb2-T cells. AB - Prolactin (PRL)-dependent rat pre-T Nb2 (Nb2-11) cell lines serve as a useful model for investigation of mechanisms underlying lactogen-mediated suppression of apoptosis. Glucocorticoids, such as dexamethasone (DEX), induce apoptosis in Nb2 11 cells; the addition of PRL abrogates the cytolytic actions of DEX in this model, presumably because of increased expression of survival genes. In the present study, we investigated whether inhibition of DEX-induced apoptosis by PRL in Nb2-T cells was accompanied by altered expression of Bcl-2 family members, mcl 1, bad or bcl-x(L) determined by Northern and immunoblot analysis. The results indicated that a 0.9 kb bcl-x(L) transcript was rapidly induced by PRL. It reached maximal levels within 2 to 4 h (>20-fold) before declining toward basal values. Similar results were obtained in primary cultures of mouse thymocytes exposed to DEX in combination with PRL. In addition to increasing its mRNA expression, PRL also increased Bcl-xL protein levels by 6 h. Moreover, the effect of PRL to increase bcl-x(L) appeared to reflect direct and indirect mechanisms, since it was attenuated by the inhibition of protein synthesis. Results from other experiments suggest that PRL signaling to bcl-x(L) expression was independent of the Jak2/Stat pathway but appeared to require activation of a Src tyrosine kinase. In contrast, while a 1.1 kb mcl-1 transcript was detected in proliferating and quiescent cells, PRL did not alter its expression at either mRNA or protein levels. Moreover, neither bad mRNA nor its protein product were detectable under any of the experimental conditions evaluated. We have concluded that bad and mcl-1 are unlikely candidates for apoptosis regulatory genes modulated by PRL. However, the kinetic pattern of PRL-provoked bcl-x(L) expression is consistent with its playing a role as an apoptosis suppressor in Nb2-T cells and primary cultures of mouse thymocytes exposed to glucocorticoids. PMID- 12904175 TI - Development of an RIA for salmon 41 kDa IGF-binding protein. AB - Salmon plasma contains at least three IGF-binding proteins (IGFBPs) with molecular masses of 41, 28 and 22 kDa. The 41 kDa IGFBP is similar to mammalian IGFBP-3 in size, type of glycosylation and physiological responses. In this study, we developed an RIA for the 41 kDa IGFBP. The 41 kDa IGFBP purified from serum was used for antibody production and as an assay standard. Binding of three different preparations of tracer were examined: (125)I-41 kDa IGFBP, (125)I-41 kDa IGFBP cross-linked with IGF-I and 41 kDa IGFBP cross-linked with (125)I-IGF-I (41 kDa IGFBP/(125)I-IGF-I). Only binding of 41 kDa IGFBP/(125)I-IGF-I was not affected by added IGFs, and therefore it was chosen for the tracer in the RIA. Plasma 41 kDa IGFBP levels measured by RIA were increased by GH treatment (178.9+/-4.9 ng/ml) and decreased after fasting (95.0+/-7.0 ng/ml). The molarities of plasma 41 kDa IGFBP and total IGF-I were comparable, and they were positively correlated, suggesting that salmon 41 kDa IGFBP is a main carrier of circulating IGF-I in salmon, as is mammalian IGFBP-3 in mammals. During the parr smolt transformation (smoltification) of coho salmon, plasma 41 kDa IGFBP levels showed a transient peak (182.5+/-10.3 ng/ml) in March and stayed relatively constant thereafter, whereas IGF-I showed peak levels in March and April. Differences in the molar ratio between 41 kDa IGFBP and IGF-I possibly influence availability of IGF-I in the circulation during smoltification. PMID- 12904176 TI - Expression and cellular localization of insulin-like growth factor-II protein and mRNA in Sparus aurata during development. AB - The spatial localization of IGF-II protein and mRNA was investigated during larval and postlarval developmental stages of the gilthead sea bream (Sparus aurata) by immunohistochemistry and in situ hybridization, using specific antisera and riboprobes. Steady-state levels of IGF-II mRNA in larvae were determined by Northern blot analysis and were found to be increased. Immunoreactivity towards IGF-II was found in larval skin, muscle, gills, gut, olfactory epithelium and kidney. After metamorphosis, the strongest immunoreactivity was found in red skeletal muscle. Positive reaction with IGF-II antibodies was also found in the olfactory epithelium and in the epithelia of pharynx, oesophagus, stomach and kidney. In the adult, the most intense signal was observed in the red and pink musculature and in heart musculature. Immunostaining was also found in saccus vasculosus, thymus, spleen and ovary. IGF II mRNA was detected by in situ hybridization in the brain, olfactory epithelium, eye, pharynx, skeletal musculature and liver. The spatial distribution of IGF-II shown in this study is consistent with previous findings on the cellular localization of IGF type 1 receptor in the sea bream and supports a role for IGF II during development and growth of sea bream. Furthermore, these results suggest that IGF-II acts in an autocrine/paracrine manner. PMID- 12904177 TI - Activating point mutations in cyclin-dependent kinase 4 are not seen in sporadic pituitary adenomas, insulinomas or Leydig cell tumours. AB - Cell cycle dysregulation is one of the defining features of cancer. Cyclin dependent kinase 4 (CDK4), together with its regulatory subunit cyclin D, governs cell cycle progression through the G1 phase. Cyclin-dependent kinase inhibitors, including p16(INK4A) (encoded by CDKN2A), in turn regulate CDK4. In particular, dysregulation of the p16/CDK4/cyclin D complex has been established in a variety of types of human tumours. Dominant activating mutations affecting codon 24 of the CDK4 gene (replacement of Arg24 by Cys or His) render CDK4 insensitive to p16(INK4) inhibition and are responsible for melanoma susceptibility in some kindreds. However, 'knock-in' mice homozygous for the CDK4(R24C) mutation were noted to develop multiple neoplasia, most commonly including endocrine tumours: pituitary adenomas, insulinomas and Leydig cell testicular tumours. We therefore speculated that sporadic human endocrine tumours might also harbour such mutations. The aim of the current study was to analyze the CDK4 gene for the two characterized activating mutations, R24C and R24H, in sporadic human pituitary adenomas, insulinomas and Leydig cell tumours. We used DNA extracted from 61 pituitary adenomas, and paired tumorous and neighboring normal genomic DNA extracted from 14 insulinoma and 6 Leydig cell tumour samples. Genomic DNA from patients with familial melanoma harbouring the R24C or the R24H mutations served as positive controls. All samples were subjected to PCR, mutation-specific restriction digests and/or sequencing. Both methodologies failed to detect mutations at these two sites in any of the sporadic endocrine tumours including pituitary adenomas, benign or malignant insulinomas or Leydig cell tumours, while the positive controls showed the expected heterozygote patterns. Protein expression of CDK4 was demonstrated by immunohistochemistry and Western blotting in pituitary and pancreatic samples. These data suggest that the changes in the regulatory 'hot-spot' on the CDK4 gene, causing various endocrine tumours in CDK4(R24C/R24C )mice, are not a major factor in sporadic pituitary, insulin beta cell or Leydig cell tumorigenesis. PMID- 12904178 TI - Developmental changes in gonadotropin-inhibitory hormone in the Japanese quail (Coturnix japonica) hypothalamo-hypophysial system. AB - We previously isolated a novel dodecapeptide containing a C-terminal -Arg-Phe NH(2) sequence, SIKPSAYLPLRF-NH(2) (RFamide peptide), from the Japanese quail (Coturnix japonica) brain. This novel quail peptide was shown to be located in neurons of the paraventricular nucleus (PVN) and their terminals in the median eminence (ME), and to decrease gonadotropin release from cultured anterior pituitary in adult birds. We therefore designated this peptide gonadotropin inhibitory hormone (GnIH). Furthermore, a cDNA encoding the GnIH precursor polypeptide has been characterized. To understand the physiological roles of this peptide, in the present study we analyzed developmental changes in the expressions of GnIH precursor mRNA and the mature peptide GnIH during embryonic and posthatch ages in the quail diencephalon including the PVN and ME. GnIH precursor mRNA was expressed in the diencephalon on embryonic day 10 (E10) and showed a significant increase on E17, just before hatch. GnIH was also detected in the diencephalon on E10 and increased significantly around hatch. Subsequently, the diencephalic GnIH content decreased temporarily, and again increased progressively until adulthood. GnIH-like immunoreactive (GnIH-ir) neurons were localized in the PVN on E10, but GnIH-ir fibers did not extend to the ME. However, GnIH-ir neurons increased in the PVN on E17, just before hatch, and GnIH-ir fibers extended to the external layer of the ME, as in adulthood. These results suggest that GnIH begins its function around hatch and acts as a hypothalamic factor to regulate gonadotropin release in the bird. PMID- 12904179 TI - Both estrogen and raloxifene cause G1 arrest of vascular smooth muscle cells. AB - The proliferation of vascular smooth muscle cells (VSMC) is a crucial pathophysiological process in the development of atherosclerosis. Although estrogen is known to inhibit the proliferation of VSMC, the mechanism responsible for this effect remains to be elucidated. In addition, the effect of raloxifene on VSMC remains unknown. We have shown here that 17beta-estradiol (E(2)) and raloxifene significantly inhibited the platelet-derived growth factor (PDGF) stimulated proliferation of cultured human VSMC. Flow cytometry demonstrated that PDGF-stimulated S-phase progression of the cell cycle in VSMC was also suppressed by E(2) or raloxifene. We found that PDGF-induced phosphorylation of retinoblastoma protein (pRb), whose hyperphosphorylation is a hallmark of the G1 S transition in the cell cycle, was significantly inhibited by E(2) and raloxifene. These effects were associated with a decrease in cyclin D1 expression, without a change in cyclin-dependent kinase 4 or cyclin-dependent kinase inhibitor, p27(kip1) expression. ICI 182,780 abolished the inhibitory effects of E(2) and raloxifene on PDGF-induced pRb phosphorylation. Next, we examined which estrogen receptor (ER) is necessary for these effects of E(2) and raloxifene. Since VSMC express both ERalpha and ERbeta, A10, a rat aortic smooth muscle cell line that expresses ERbeta but not ERalpha, was used. The dose dependent stimulation of A10 cell proliferation by PDGF was not inhibited by E(2) or raloxifene in contrast to the results obtained in VSMC. Moreover, E(2) and raloxifene significantly inhibited the PDGF-induced cyclin D1 promoter activity in A10 cells transfected with cDNA for ERalpha but not in the parental cells. These results suggested that E(2) and raloxifene exert an antiproliferative effect in VSMC treated with PDGF, at least in part through inhibition of pRb phosphorylation, and that the inhibitory effects of E(2) and raloxifene may be mainly mediated by ERalpha. PMID- 12904180 TI - Implication of lipids in macrosomia of diabetic pregnancy: can n-3 polyunsaturated fatty acids exert beneficial effects? AB - Macrosomia or fetal obesity is a frequent complication of pregnancy in diabetes mellitus. Several alterations observed in carbohydrate and lipid metabolism in macrosomic infants of diabetic mothers are thought to be a consequence of maternal hyperglycaemia leading to fetal hyperinsulinaemia. Macrosomic infants of diabetic mothers are prone to the development of glucose intolerance, obesity and diabetes during childhood and adulthood. Furthermore, increasing evidence is accumulating regarding the importance of n -3 polyunsaturated fatty acids (PUFAs) in the reduction of plasma lipids and hyperglycaemia. In this review article, we shed light on the abnormalities in lipid metabolism in macrosomia. We also raise the question of the possible beneficial effects of n -3 PUFAs in diabetic pregnancy and in the prevention and treatment of long-term metabolic abnormalities associated with macrosomia. PMID- 12904181 TI - Structure and transcriptional regulation of the Nat2 gene encoding for the drug metabolizing enzyme arylamine N-acetyltransferase type 2 in mice. AB - Arylamine N-acetyltransferases (NATs) are polymorphic enzymes, well-known for their role in the metabolism of drugs and carcinogens. Mice have three NAT isoenzymes, of which NAT2 is postulated to be involved in endogenous, as well as xenobiotic, metabolism. To understand expression of the murine Nat2 gene, we have analysed its structure and transcriptional regulation. We have accurately mapped the transcription initiation site 6.5 kb upstream of the coding region of the gene, adjacent to a recently described non-coding exon. Transcription was demonstrated to start from this region in embryonic and adult liver, spleen, submaxillary gland, kidney, brain, thymus, lung and placenta, but not in the heart. Database searches and analyses of cDNA by PCR suggested alternative splicing of the single 6.2 kb intron of Nat2, and determined the position of the polyadenylation signal at 0.44 kb downstream of the coding region of the gene. Examination of the 13 kb sequence flanking the coding and non-coding exons of Nat2 revealed a single promoter, located close to the transcription-initiation site, and indicated regions likely to harbour control elements. The Nat2 promoter consists of an atypical TATA box and a Sp1 [SV40 (simian virus 40) protein 1] box identical with that found in many housekeeping gene promoters. Activity of the Nat2 promoter was severely reduced by deletion or mutation of either of these two elements, whereas the region of the Sp1 box bound cellular protein and resisted DNase I digestion. Finally, the ability of the promoter region to bind cellular protein was reduced by competition with oligonucleotides bearing the Sp1 consensus sequence. PMID- 12904182 TI - Sevoflurane: an ideal agent for adult day-case anesthesia? AB - Sevoflurane has several properties which make it potentially useful as a day case anaesthetic. Following induction of anaesthesia with propofol, awakening from sevoflurane is faster compared to isoflurane, faster or similar compared to propofol and comparable (in the majority of studies) to desflurane. Subsequent recovery and discharge is generally similar following all agents. Sevoflurane may also be used to induce anaesthesia, which is generally well-received and causes less hypotension and apnoea compared to propofol. When used as a maintenance anaesthetic, the incidence of postoperative nausea and vomiting after sevoflurane is comparable to other inhaled anaesthetics, but this complication appears more common after inhaled inductions. The tolerability and low solubility of sevoflurane facilitate titration of anaesthesia and may reduce the need for opioid analgesia, which in turn may limit the occurrence of nausea and vomiting. PMID- 12904183 TI - Effects of 0.5 and 1.0 MAC isoflurane, sevoflurane and desflurane on intracranial and cerebral perfusion pressures in children. AB - BACKGROUND: Isoflurane has been a commonly used agent for neuroanesthesia, but newer agents, sevoflurane and desflurane, have a quicker onset and shorter emergence from anesthesia and are increasingly preferred for general pediatric anesthesia. But their effects on intracranial pressure (ICP) and cerebral perfusion pressure (CPP), especially in pediatric patients with already increased ICP, have not been well documented. METHODS: We studied 36 children scheduled for elective implantation of an intraparenchymal pressure device for 24 h monitoring for suspected elevated ICP. After a standardized intravenous anesthesia, the patients were moderately hyperventilated with 60% nitrous oxide (N2O) in oxygen. The patients were then randomized to receive 0.5 and 1.0 MAC of isoflurane (Group I, n = 12), sevoflurane (Group S, n = 12) or desflurane (Group D, n = 12) in 60% N2O in oxygen. Respiratory and hemodynamic variables, ICP and CPP were recorded at baseline and after exposure to a target level of test drug for 10 min or until CPP fell below 30 mmHg (recommended lower ICP level is 25 mmHg in neonates, rising to 40 mmHg in toddlers). RESULTS: When comparing baseline values with values at 1.0 MAC, mean arterial pressure (MAP) decreased (P < 0.001) in all groups, with no differences between the groups. ICP increased (P < 0.001) with all agents, mean +2, +5, and +6 mmHg in Group I, S and D, respectively, with no differences between the groups. Regression analyzes found no relationship between baseline ICP and the increases in ICP from baseline to 1.0 MAC for isoflurane or sevoflurane. However, increased baseline ICP tended to cause a higher ICP increase with 1.0 MAC desflurane; regression coefficient +0.759 (P = 0.077). The difference between regression coefficients for Group I and Group D were not significant (P = 0.055). CPP (MAP-ICP) decreased (P < 0.001) in all groups, mean 18, -14 and -17 mmHg in Group I, S and D, respectively, with no significant difference between the groups. CONCLUSIONS: 0.5 and 1.0 MAC isoflurane, sevoflurane and desflurane in N2O all increased ICP and reduced MAP and CPP in a dose-dependent and clinically similar manner. There were no baseline dependent increases in ICP from 0 to 1.0 MAC with isoflurane or sevoflurane, but ICP increased somewhat more, although statistically insignificant, with higher baseline values in patients given desflurane. The effect of MAP on CPP is 3-4 times higher than the effect of the increases in ICP on CPP and this makes MAP the most important factor in preserving CPP. In children with known increased ICP, intravenous anesthesia may be safer. However, maintaining MAP remains the most important determinant of a safe CPP. PMID- 12904184 TI - Relaxing intraoperative natural sound blunts haemodynamic change at the emergence from propofol general anaesthesia and increases the acceptability of anaesthesia to the patient. AB - BACKGROUND: It is known that auditory input, such as comforting music or sound, blunts the human response to surgical stress in conscious patients under regional anaesthesia. As auditory perception has been demonstrated to remain active under general anaesthesia, playing comforting sounds to patients under general anaesthesia might also modulate the response of these patients to surgical stress. METHODS: Fifty-nine patients scheduled for laparoscopic cholecystectomy were anaesthetized with propofol general anaesthesia in combination with epidural anaesthesia. Natural sounds, chosen preoperatively by each patient as being comforting, were played to 29 patients using headphones during surgery (S group) and the remainder of the patients (n = 30) were fitted with dummy open-type headphones (N group). We compared the haemodynamic change during anaesthesia and the acceptability of anaesthetic practice between the two groups in a randomized double-blind design. RESULTS: There were no differences in haemodynamics between the S and N groups during surgery. During the emergence from anaesthesia, the mean blood pressure and heart rate gradually increased; both parameters were significantly higher in the N group than in the S group. Postoperatively, patients in the S group perceived the experience of anaesthesia as significantly more acceptable than did those in the N group. CONCLUSION: These findings indicate that allowing patients comforting background sounds during general anaesthesia may blunt haemodynamic changes upon emergence from general anaesthesia and increase the acceptability of the experience of anaesthesia. PMID- 12904185 TI - A new system to target the effect-site during propofol sedation. AB - BACKGROUND: We evaluated a new, integrated, covariate-adjusted, target-controlled infusion system during sedation with propofol combined with 50% nitrous oxide (N2O) and with propofol only (Air). METHODS: The protocol consisted of sequential 15-minute cycles in 20 volunteers. After a 15-minute control period, propofol was infused to an initial target effect-site concentration of 0.25 microg x ml-1 (N2O) or 1.5 microg x ml-1 (Air). Subsequently, the target effect-site concentration was increased by 0.25 (N2O) or 0.5 microg x ml-1 (Air) for 15 min This sequence was continued until the volunteers lost consciousness as defined by an Observer's Assessment Alertness/Sedation (OAA/S) score = 2. RESULTS: Venous plasma propofol concentrations at the beginning(9 elapsed minutes) and end(15 elapsed minutes) of the pseudo-steady state period differed by only 0.00 +/- 0.16 microg x ml-1 (P = 0.78) during the N2O and 0.00 +/- 0.25 microg x ml-1 (P = 0.91) during the Air trial. OAA/S scores and bispectral index values, as surrogate measures of pharmacodynamic effect, were not different during this time in either trial. The median(25th, 75th percentiles) of the median performance error (%) was -13 (-24, -1) during the N2O and -18 (-26, -9) during the Air trial. The median absolute performance error (%) was 17 (10, 24) in the N2O and 22 (12, 28) in Air trial. The divergence (%/h) was -10 (-26, 4) in the N2O and 14 (-21, 26) in Air trial. The wobble was 7 (5, 10) in the N2O and 6 (4, 8) in the Air trial. CONCLUSIONS: When tested with venous blood samples, our TCI system for propofol, using a covariate-adjusted, integrated pharmacokinetic model to target effect-site concentrations, demonstrated a clinically acceptable accuracy and stability during mild to moderate sedation. PMID- 12904186 TI - Automated responsiveness test and bispectral index monitoring during propofol and propofol/N2O sedation. AB - BACKGROUND: Sedation practice, especially when non-anaesthesia personnel are involved, requires efficient anaesthetic depth monitoring. Therefore, we used prediction probability (PK) to evaluate the performance of the bispectral index (BIS) of the EEG and automated responsiveness test (ART) to predict sedation depth and loss of subject's responsiveness during propofol sedation, with and without N2O. METHODS: Twenty volunteers were studied during propofol administration with (N2O) and without (Air) N2O. The protocol consisted of sequential 15-min cycles. After a control period, propofol was infused to a target effect-site concentration of 0.25 microg/ml (N2O) or 1.5 microg/ml (Air), which was subsequently increased by 0.25 or 0.5 microg/ml, respectively, until loss of responsiveness was detected by loss of response to command [observer's assessment of alertness/sedation (OAA/S) score 2'-F > 2'-N > 2' H. Effects on the rate of the chemical step (about 10-fold for 2'-F, almost 150 fold for 2'-H and 2'-N) were much stronger, and, except for the 2'-N modification, resembled strikingly those observed in the Tetrahymena ribozyme catalyzed reaction at corresponding position. Mn2+ rescued cleavage of the 2'-N but also the 2'-H-modified ptRNA, arguing against a direct metal ion coordination at this location. Miscleavage between nt -1 and -2 was observed for the 2'-N ptRNA at low pH (further influenced by the base identities at nt -1 and +73), suggesting repulsion of a catalytic metal ion due to protonation of the amino group. Effects caused by the 2'-N modification at nt -1 of the substrate allowed us to substantiate a mechanistic difference in phosphodiester hydrolysis catalyzed by Escherichia coli RNase P RNA and the Tetrahymena ribozyme: a metal ion binds next to the 2' substituent at nt -1 in the reaction catalyzed by RNase P RNA, but not at the corresponding location in the Tetrahymena ribozyme reaction. PMID- 12904301 TI - Regulation of a TRPM7-like current in rat brain microglia. AB - Non-excitable cells use Ca2+ influx for essential functions but usually lack voltage-gated Ca2+ channels. The main routes of Ca2+ entry appear to be store operated channels or Ca2+-permeable non-selective cation channels, of which the magnesium-inhibited cation (or magnesium-nucleotide-regulated metal cation) current has received considerable recent attention. This current appears to be produced by one of the recently cloned transient receptor potential (TRP) channels, TRPM7. In this study of rat microglia, we identified TRPM7 transcripts and a prevalent current with the hallmark biophysical and pharmacological features of TRPM7. This is the first identification of a TRPM7-like current in the brain. There is little known about how members of the TRPM sub-family normally become activated. Using whole-cell patch clamp recordings from rat microglia, we found that the TRPM7-like current activates spontaneously after break-in and that the current and its activation are inhibited by elevated intracellular Mg2+ but not affected by cell swelling or a wide range of intracellular Ca2+ concentrations. The TRPM7-like current in microglia appears to depend on tyrosine phosphorylation. It was inhibited by several tyrosine kinase inhibitors, including a peptide (Src 40-58) that was shown previously to inhibit Src actions, but not by inactive drugs or peptide analogues. The current did not depend on the cell activation state; i.e. it was the same in microglia recently removed from the brain or when cultured under a wide range of conditions that favor the resting or activated state. Because TRPM7 channels are permeable to Ca2+, this current may be important for microglia functions that depend on elevations in intracellular Ca2+. PMID- 12904302 TI - Targeted deletion of CD44v7 exon leads to decreased endothelial cell injury but not tumor cell killing mediated by interleukin-2-activated cytolytic lymphocytes. AB - In the current study, we investigated the nature and role of CD44 variant isoforms involved in endothelial cell (EC) injury and tumor cell cytotoxicity mediated by IL-2-activated killer (LAK) cells. Treatment of CD44 wild-type lymphocytes with IL-2 led to increased gene expression of CD44 v6 and v7 variant isoforms and to significant induction of vascular leak syndrome (VLS). CD44v6-v7 knockout (KO) and CD44v7 KO mice showed markedly reduced levels of IL-2-induced VLS. The decreased VLS in CD44v6-v7 KO and CD44v7 KO mice did not result from differential activation and expansion of CD8+ T cells, NK, and NK-T cells or from altered degree of perivascular lymphocytic infiltration in the lungs. LAK cells from CD44v7 KO mice showed a significant decrease in their ability to adhere to and mediate lysis of EC but not lysis of P815 tumor cells in vitro. CD44v7 mediated lysis of EC by LAK cells was dependent on the activity of phosphatidylinositol 3-kinase and tyrosine kinases. Interestingly, IL-2-activated LAK cells expressing CD44hi but not CD44lo were responsible for EC lysis. Furthermore, lysis of EC targets could be blocked by addition of soluble or enzymatic cleavage of CD44v6-v7-binding glycosaminoglycans. Finally, anti-CD44v7 mAbs caused a significant reduction in the adherence to and killing of EC and led to suppression of IL-2-induced VLS. Together, this study suggests that the expression of CD44v7 on LAK cells plays a specific role in EC injury and that it may be possible to reduce EC injury but not tumor cell killing by specifically targeting CD44v7. PMID- 12904303 TI - Single mutation in the linker domain confers protein flexibility and camptothecin resistance to human topoisomerase I. AB - DNA topoisomerase I relaxes supercoiled DNA by the formation of a covalent intermediate in which the active-site tyrosine is transiently bound to the cleaved DNA strand. The antineoplastic agent camptothecin specifically targets DNA topoisomerase I, and several mutations have been isolated that render the enzyme camptothecin-resistant. The catalytic and structural dynamical properties of a human DNA topoisomerase I mutant in which Ala-653 in the linker domain was mutated into Pro have been investigated. The mutant is resistant to camptothecin and in the absence of the drug displays a cleavage-religation equilibrium strongly shifted toward religation. The shift is mainly because of an increase in the religation rate relative to the wild type enzyme, indicating that the unperturbed linker is involved in slowing religation. Molecular dynamics simulation indicates that the Ala to Pro mutation increases the linker flexibility allowing it to sample a wider conformational space. The increase in religation rate of the mutant, explained by means of the enhanced linker flexibility, provides an explanation for the mutant camptothecin resistance. PMID- 12904304 TI - Activation of the Rap GTPases in B lymphocytes modulates B cell antigen receptor induced activation of Akt but has no effect on MAPK activation. AB - Signaling by the B cell antigen receptor (BCR) activates the Rap1 and Rap2 GTPases, putative antagonists of Ras-mediated signaling. Because Ras can activate the Raf-1/ERK pathway and the phosphatidylinositol 3-kinase (PI3K)/Akt pathway, we asked whether Rap activation limits the ability of the BCR to signal via these pathways. To do this, we blocked the activation of endogenous Rap1 and Rap2 by expressing the Rap-specific GTPase-activating protein RapGAPII. Preventing Rap activation had no effect on BCR-induced activation of ERK. In contrast, BCR induced phosphorylation of Akt on critical activating sites was increased 2- to 3 fold when Rap activation was blocked. Preventing Rap activation also increased the ability of the BCR to stimulate Akt-dependent phosphorylation of the FKHR transcription factor on negative regulatory sites and decreased the levels of p27Kip1, a pro-apoptotic factor whose transcription is enhanced by FKHR. Moreover, preventing Rap activation reduced BCR-induced cell death in the WEHI 231 B cell line. Thus activation of endogenous Rap by the BCR limits BCR-induced activation of the PI3K/Akt pathway, opposes the subsequent inhibition of the FKHR/p27Kip1 pro-apoptotic module, and enhances BCR-induced cell death. Consistent with the idea that Rap-GTP is a negative regulator of the PI3K/Akt pathway, expressing constitutively active Rap2 (Rap2V12) reduced BCR-induced phosphorylation of Akt and FKHR. Finally, our finding that Rap2V12 can bind PI3K and inhibit its activity in a manner that depends upon BCR engagement provides a potential mechanism by which Rap-GTP limits activation of the PI3K/Akt pathway, a central regulator of B cell growth and survival. PMID- 12904305 TI - Tissue inhibitor of metalloproteinase-1 protects human breast epithelial cells against intrinsic apoptotic cell death via the focal adhesion kinase/phosphatidylinositol 3-kinase and MAPK signaling pathway. AB - Tissue inhibitor of metalloproteinase (TIMP-1) is a natural protease inhibitor of matrix metalloproteinases (MMPs). Recent studies revealed a novel function of TIMP-1 as a potent inhibitor of apoptosis in mammalian cells. However, the mechanisms by which TIMP-1 exerts its anti-apoptotic effect are not understood. Here we show that TIMP-1 activates cell survival signaling pathways involving focal adhesion kinase, phosphatidylinositol 3-kinase, and ERKs in human breast epithelial cells to TIMP-1. TIMP-1-activated cell survival signaling down regulates caspase-mediated classical apoptotic pathways induced by a variety of stimuli including anoikis, staurosporine exposure, and growth factor withdrawal. Consistently, down-regulation of TIMP-1 expression greatly enhances apoptotic cell death. In a previous study, substitution of the second amino acid residue threonine for glycine in TIMP-1, which confers selective MMP inhibition, was shown to obliterate its anti-apoptotic activity in activated hepatic stellate cells suggesting that the anti-apoptotic activity of TIMP-1 is dependent on MMP inhibition. Here we show that the same mutant inhibits apoptosis of human breast epithelial cells, suggesting different mechanisms of TIMP-1 regulation of apoptosis depending on cell types. Neither TIMP-2 nor a synthetic MMP inhibitor protects breast epithelial cells from intrinsic apoptotic cell death. Furthermore, TIMP-1 enhances cell survival in the presence of the synthetic MMP inhibitor. Taken together, the present study unveils some of the mechanisms mediating the anti-apoptotic effects of TIMP-1 in human breast epithelial cells through TIMP-1-specific signal transduction pathways. PMID- 12904306 TI - Estrogens down-regulate p27Kip1 in breast cancer cells through Skp2 and through nuclear export mediated by the ERK pathway. AB - The cyclin-dependent kinase (CDK) inhibitor p27Kip1 plays a key role in growth and development of the mammary epithelium and in breast cancer. p27Kip1 levels are regulated through ubiquitin/proteasome-mediated proteolysis, promoted by CDK2 and the F box protein Skp2 at the G1/S transition, and independent of Skp2 in mid G1. We investigated the respective roles of Skp2 and subcellular localization of p27Kip1 in down-regulation of p27Kip1 induced in MCF-7 cells by estrogens. 17beta Estradiol treatment increased Skp2 expression in MCF-7 cells; however, this increase was prevented by G1 blockade mediated by p16Ink4a or the CDK inhibitor roscovitine, whereas down-regulation of p27Kip1 was maintained. Exogenous Skp2 prevented growth arrest of MCF-7 cells by antiestrogen, coinciding with decreased p27Kip1 expression. Under conditions of G1 blockade, p27Kip1 was stabilized by inhibition of CRM1-dependent nuclear export with leptomycin B or by mutation of p27Kip1 (Ser10 --> Ala; S10A) interfering with CRM1/p27Kip1 interaction. Antisense Skp2 oligonucleotides and a dominant-interfering Cul-1(1-452) mutant prevented down-regulation of p27Kip1S10A, whereas Skp2 overexpression elicited its destruction in mitogen-deprived cells. Active mediators of the extracellular signal-regulated kinase (ERK) pathway including Raf-1caax induced cytoplasmic localization of p27Kip1 in antiestrogen-treated cells and prevented accumulation of p27Kip1 in these cells independent of Skp2 expression and coinciding with ERK activation. Genetic or chemical blockade of the ERK pathway prevented down regulation and cytoplasmic localization of p27Kip1 in response to estrogen. Our studies indicate that estrogens elicit down-regulation of p27Kip1 in MCF-7 cells through Skp2-dependent and -independent mechanisms that depend upon subcellular localization of p27Kip1 and require the participation of mediators of the Ras/Raf 1/ERK signaling pathway. PMID- 12904307 TI - Aminophospholipids have no access to the luminal side of the biliary canaliculus: implications for thr specific lipid composition of the bile fluid. AB - About 95% of the bile phospholipids are phosphatidylcholine. Although the fractions of phosphatidylcholine and of both aminophospholipids phosphatidylserine and phosphatidylethanolamine in the canalicular membrane are in the same order of about 35% of total lipids, both aminophospholipids are almost absent from the bile. To rationalize this observation, we studied the intracellular uptake of various fluorescent phospholipid analogues and their subsequent enrichment in the bile canaliculus (BC) of HepG2 cells. Diacylaminophospholipid analogues but not phosphatidylcholine analogues became rapidly internalized by an aminophospholipid translocase (APLT) activity in the plasma membrane of HepG2 cells. We observed only low labeling of BC by diacylaminophospholipids but extensive staining by phosphatidylcholine analogues. In the presence of suramin, known to inhibit APLT, a strong labeling of BC by diacylaminophospholipid analogues was found that declined to a level observed for control cells after removal of suramin. Unlike diacylphosphatidylserine, diether phosphatidylserine analogue, which is not an appropriate substrate of APLT, accumulated in the BC. The correlation between low labeling of BC and an APLT mediated transbilayer movement suggests the presence of an APLT activity in the canalicular membrane that prevents exposure of aminophospholipids to the bile. PMID- 12904308 TI - Identification of the ZPC oligosaccharide ligand involved in sperm binding and the glycan structures of Xenopus laevis vitelline envelope glycoproteins. AB - The Xenopus laevis egg vitelline envelope is composed of five glycoproteins (ZPA, ZPB, ZPC, ZPD, and ZPX). As shown previously, ZPC is the primary ligand for sperm binding to the egg envelope, and this binding involves the oligosaccharide moieties of the glycoprotein (Biol. Reprod., 62:766-774, 2000). To understand the molecular mechanism of sperm-egg envelope binding, we characterized the N-linked glycans of the vitelline envelope (VE) glycoproteins. The N-linked glycans of the VE were composed predominantly of a heterogeneous mixture of high-mannose (5-9) and neutral, complex oligosaccharides primarily derived from ZPC (the dominant glycoprotein). However, the ZPA N-linked glycans were composed of acidic-complex and high-mannose oligosaccharides, ZPX had only high-mannose oligosaccharides, and ZPB lacked N-linked oligosaccharides. The consensus sequence for N-linked glycosylation at the evolutionarily conserved residue N113 of the ZPC protein sequence was glycosylated solely with high-mannose oligosaccharides. This conserved glycosylation site may be of importance to the three-dimensional structure of the ZPC glycoproteins. One of the complex oligosaccharides of ZPC possessed terminal beta-N-acetyl-glucosamine residues. The same ZPC oligosaccharide species isolated from the activated egg envelopes lacked terminal beta-N-acetyl-glucosamine residues. We previously showed that the cortical granules contain beta-N-acetyl-glucosaminidase (J. Exp. Zool., 235:335-340, 1985). We propose that an alteration in the oligosaccharide structure of ZPC by glucosaminidase released from the cortical granule reaction is responsible for the loss of sperm binding ligand activity at fertilization. PMID- 12904309 TI - Roles of tumor necrosis factor-alpha of the estrous cycle in cattle: an in vivo study. AB - We have suggested in a previous in vitro study that tumor necrosis factor-alpha (TNFalpha) plays a role in the initiation of luteolysis in cattle. The aim of the present study was to examine the influence of different doses of TNFalpha on the estrous cycle in cattle by observing the standing behavior and measuring peripheral concentrations of progesterone (P4) during the estrous cycle. Moreover, we evaluated the secretion of P4, oxytocin (OT), nitric oxide (NO), and luteolytic (prostaglandin F2alpha [PGF2alpha] and leukotriene C4 [LTC4]) and luteotropic (PGE2) metabolites of arachidonic acid in peripheral blood plasma as parameters of TNFalpha actions. Mature Holstein/Polish black and white heifers (n = 36) were treated on Day 14 of the estrous cycle (Day 0 = estrus) by infusion into the aorta abdominalis of saline (n = 8), an analogue of PGF2alpha (cloprostenol, 100 microg; n = 3) or saline with TNFalpha at doses of 0.1 (n = 3), 1 (n = 8), 10 (n = 8), 25 (n = 3), or 50 microg (n = 3) per animal. Peripheral blood samples were collected frequently before, during, and up to 4 h after TNFalpha treatment. After Day 15 of the estrous cycle, blood was collected once daily until Day 22 following the first estrus. Lower doses of TNFalpha (0.1 and 1 microg) decreased the P4 level during the estrous cycle and consequently resulted in shortening of the estrous cycle (18.8 +/- 0.9 and 18.0 +/- 0.7 days, respectively) compared with the control (22.3 +/- 0.3 days, P < 0.05). One microgram of TNFalpha increased the PGF2alpha (P < 0.001) and NO (P < 0.001) concentrations and decreased OT secretion (P < 0.01). Higher doses of TNFalpha (10, 25, 50 microg) stimulated synthesis of P4 (P < 0.001) and PGE2 (P < 0.001), inhibited LTC4 secreton (P < 0.05), and consequently resulted in prolongation of the estrous cycle (throughout 30 days, P < 0.05). Altogether, the results suggest that low concentrations of TNFalpha cause luteolysis, whereas high concentrations of TNFalpha activate corpus luteum function and prolong the estrous cycle in cattle. PMID- 12904310 TI - Identification of a biological activity that supports maintenance and proliferation of pluripotent cells from the primitive ectoderm of the mouse. AB - Pluripotent cell development in the mammalian embryo results in the sequential formation of several developmentally distinct populations, inner cell mass, primitive ectoderm, and the primordial germ lineage. Factors within medium conditioned by HepG2 cells (MEDII) have been implicated in the formation and maintenance of primitive ectoderm from inner cell mass cells both in vitro and in vivo. Here we demonstrate that MEDII, but not LIF, is able to support the maintenance and proliferation in culture of pluripotent cells derived from primitive ectoderm formed in vitro or during embryonic development. This distinguishes primitive ectoderm and inner cell mass (ICM) on the basis of cytokine responsiveness and validates the biological activity proposed for factors within MEDII in primitive ectoderm establishment and maintenance. Further, it potentially provides an alternative technology for the isolation of pluripotent cells from the mammalian embryo. PMID- 12904311 TI - Onset and progress of meiotic prophase in the oocytes in the B6.YTIR sex-reversed mouse ovary. AB - When the Y chromosome of a Mus musculus domesticus male mouse (caught in Tirano, Italy) is placed on a C57BL/6J genetic background, approximately half of the XY (B6.YTIR) progeny develop into normal-appearing but infertile females. We have previously reported that the primary cause of infertility can be attributed to their oocytes. To identify the primary defect in the XY oocyte, we examined the onset and progress of meiotic prophase in the B6.YTIR fetal ovary. Using bromo deoxyuridine incorporation and culture, we determined that the germ cells began to enter meiosis at the developmental ages and in numbers comparable to those in the control XX ovary. Furthermore, the meiotic prophase appeared to progress normally until the late zygotene stage. However, the oocytes that entered meiosis early in the XY ovary failed to complete the meiotic prophase. On the other hand, a considerable number of oocytes entered meiosis at late developmental stages and completed the meiotic prophase in the XY ovary. We propose that the timing of entry into meiosis and the XY chromosomal composition influence the survival of oocytes during meiotic prophase in the fetal ovary. PMID- 12904312 TI - 11beta-hydroxysteroid dehydrogenase and glucocorticoid receptor messenger RNA expression in porcine placentae: effects of stage of gestation, breed, and uterine environment. AB - Glucocorticoids are known to influence many aspects of prenatal development. Three important regulators of glucocorticoid actions at the cellular level are the enzymes 11beta-hydroxysteroid dehydrogenase type 1 (11betaHSD-1), 11beta hydroxysteroid dehydrogenase type 2 (11betaHSD-2), and glucocorticoid receptors (GR). The present study was conducted to determine the presence of these regulators in porcine placentae during early gestation (Days 24-40; term = 114 days) and to examine the influence of breed and uterine environment. Three pig models differing in uterine environment as reflected by embryonic survival from Days 24 to 40 were used: intact white cross-bred gilts (WC-INT); white cross-bred gilts that had been unilaterally hysterectomized-ovariectomized before puberty (WC-UHO); and intact Meishan gilts (ME). Porcine-specific partial cDNAs for 11betaHSD-1 and 11betaHSD-2 and a cRNA for GRalpha were developed and used to produce 32P-labeled probes for Northern blot analyses. The 11betaHSD dehydrogenase activity was measured in vitro at saturating concentrations of substrate and coenzyme. At Day 24 of gestation, 11betaHSD-2 mRNA, dehydrogenase activity, and GR mRNA were present, but 11betaHSD-1 mRNA was absent. All three mRNAs and dehydrogenase activity increased (P < 0.01) by Day 40. On Day 30, placental 11betaHSD-2 mRNA was decreased (P = 0.03) by 47% in WC-UHO versus WC INT. Placental 11betaHSD dehydrogenase activity was 2-fold greater (P < 0.01) in ME versus WC-INT on Day 24 of gestation. These results demonstrate, to our knowledge for the first time, the presence of 11betaHSD-1, 11betaHSD-2, and GR mRNA as well as 11betaHSD dehydrogenase activity in the porcine placenta during early pregnancy. Moreover, a role for glucocorticoids in porcine embryonic development is suggested. PMID- 12904313 TI - Effects of in vitro production on horse embryo morphology, cytoskeletal characteristics, and blastocyst capsule formation. AB - Blastocyst formation rates during horse embryo in vitro production (IVP) are disappointing, and embryos that blastulate in culture fail to produce the characteristic and vital glycoprotein capsule. The aim of this study was to evaluate the impact of IVP on horse embryo development and capsule formation. IVP embryos were produced by intracytoplasmic sperm injection of in vitro matured oocytes and either culture in synthetic oviduct fluid (SOF) or temporary transfer to the oviduct of a ewe. Control embryos were flushed from the uterus of mares 6 9 days after ovulation. Embryo morphology was evaluated with light microscopy, and multiphoton scanning confocal microscopy was used to examine the distribution of microfilaments (AlexaFluor-Phalloidin stained) and the rate of apoptosis (cells with fragmented or terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling-positive nuclei). To examine the influence of culture on capsule formation, conceptuses were stained with a monoclonal antibody specific for capsular glycoproteins (OC-1). The blastocyst rate was higher for zygotes transferred to a sheep's oviduct (16%) than for those cultured in SOF (6.3%). Day 7 IVP embryos were small and compact with relatively few cells, little or no blastocoele, and an indistinct inner cell mass. IVP embryos had high percentages of apoptotic cells (10% versus 0.3% for in vivo embryos) and irregularly distributed microfilaments. Although they secreted capsular glycoproteins, the latter did not form a normal capsule but instead permeated into the zona pellucida or remained in patches on the trophectodermal surface. These results demonstrate that the initial layer of capsule is composed of OC-1-reactive glycoproteins and that embryo development ex vivo is retarded and aberrant, with capsule formation failing as a result of failed glycoprotein aggregation. PMID- 12904314 TI - Effects of mono-(2-ethylhexyl) phthalate on fetal and neonatal rat testis organ cultures. AB - Di-(2-ethylhexyl) phthalate (DEHP) and its active metabolite, mono-(2-ethylhexyl) phthalate (MEHP), have been shown to cause reproductive toxicity in both developing and adult animals. In this study, we used organ cultures of fetal and neonatal rat testes to assess the in vitro effect of MEHP on seminiferous cord formation in Embryonic Day 13 (E13) testes and on the development of E18 and Postnatal Day 3 (P3) testes. Interestingly, MEHP had no effect on cord formation in the organ cultures of E13 testes, indicating that it has no effect on sexual differentiation of the indifferent gonad to testis. Consistently, the expression of a Sertoli cell-specific protein, mullerian inhibiting substance (MIS), or the number of gonocytes did not change in E13 testes after MEHP treatment. In contrast, MEHP decreased the levels of MIS and GATA-4 proteins in Sertoli cells and impaired Sertoli cell proliferation in the organ cultures of E18 and P3 testes. These results suggest that MEHP negatively influences proliferation and differentiation of Sertoli cells in both fetal and neonatal testes. In addition, MEHP treatment did not alter the number of gonocytes in E18 testes, whereas the number of gonocytes in P3 testes decreased in a dose-dependent manner, apparently due to enhanced apoptosis. These results suggest that MEHP adversely affects the gonocytes, which are mitotically active and undergoing migration and differentiation in neonatal testes, but it has no effect on fetal gonocytes that are mitotically quiescent. PMID- 12904315 TI - Reprogramming of bovine somatic cell nuclei is not directly regulated by maturation promoting factor or mitogen-activated protein kinase activity. AB - Cloned mammals with normal fertility have been produced by nuclear transfer. Thus, oocyte cytoplasm has the ability to convert differentiated somatic cell nuclei into a state that resembles the conditions that occur at fertilization (nuclear reprogramming). Despite the long-held assumption that reprogramming factors are present in mammalian oocytes, the molecular nature of these factors is not known. The present study demonstrates that the process of nuclear reprogramming is not directly regulated by maturation promoting factor or mitogen activated protein kinase activity. The potential for nuclear-transferred oocytes to develop to the blastocyst stage was not different when somatic cells at the M phase were fused with oocytes activated with ionomycin and cycloheximide 1-5 h before (12%-22%) but was significantly decreased when oocytes were activated 6 h before (1%). Further molecular studies on the differences between oocytes with and without reprogramming potential are required and will be useful for the identification of reprogramming factors. PMID- 12904316 TI - Calcium current subtypes in GnRH neurons. AB - Calcium plays roles in excitability, rhythm generation, and neurosecretion. Identifying channel subtypes that regulate calcium influx is thus important to understanding rhythmic GnRH secretion, which is a prerequisite for reproduction. Whole-cell voltage-clamp recordings were made from short-term dissociated GnRH adult ovariectomized (OVX) mice (n = 21) to identify channel subtypes that carry calcium current using selective channel blockers and voltage characteristics. Low voltage activated (LVA) currents were not observed in 42 GnRH neurons tested, although most non-GnRH neurons (4/6) displayed LVA current. The L-type component of the high-voltage activated (HVA) calcium current was 25% +/- 2%. The remaining HVA calcium current passed through N-type (27% +/- 3%), P-type (15% +/- 1%), Q type (18% +/- 3%), and R-type (15% +/- 1%) channels. Because these data differ substantially from reports on cultured GnRH neurons, which may represent reproductively immature models, we also examined GnRH neurons from gonadal-intact young (Postnatal Days 4-10, n = 8 mice) mice. LVA currents were still rare (2/28) in young mice. Although the same HVA components were observed, the proportions were shifted toward significantly more L-type and less N-type current, suggesting a possible developmental shift in calcium currents in GnRH neurons. These data suggest that calcium channel subtypes in GnRH neurons prepared in the short term from brain slices differ substantially from those in long-term cultured GnRH models. These findings provide a vital foundation to examine the role of calcium channels in the secretory and rhythmic machinery of GnRH neurons. PMID- 12904317 TI - Genetic analysis of the clonal origin of regenerating mouse spermatogenesis following transplantation. AB - Spermatogonial transplantation provides a straightforward approach to quantify spermatogonial stem cells (SSCs). Because donor-derived spermatogenesis is regenerated in the form of distinct colonies, the number of functional SSCs can be obtained by simply counting the number of colonies established in recipient testes. However, this approach is legitimate only when one colony arises from one stem cell (one colony-one stem cell hypothesis). In this study, we evaluated the validity of this hypothesis. Two populations of donor cells were obtained from the testes of two transgenic mouse lines and mixed at a 1:1 ratio. Following transplantation of the cell mixture, donor-derived colonies were visualized and individually excised, and genomic DNA was extracted from each colony. Based on unique marker genes of the two transgenic lines, the genotype of the cells contained in a colony was examined by polymerase chain reaction. A colony was determined to be clonal when only one transgene was detected. The results showed that 100% and 90% of colonies were clonal when <5 and 19 colonies were formed per recipient testis, respectively. However, the clonality of colonies decreased as the colony number per recipient testis or the length of each colony increased. These results support the one colony-one stem cell hypothesis and demonstrate that spermatogonial transplantation provides a highly quantitative assay for SSCs; however, these conclusions are applicable under a defined transplantation condition. PMID- 12904318 TI - Cyclic adenosine monophosphate (cAMP) stimulation of the kit ligand promoter in sertoli cells requires an Sp1-binding region, a canonical TATA box, and a cAMP induced factor binding to an immediately downstream GC-rich element. AB - Expression of Kit ligand (KL) mRNA is induced in primary prepuberal Sertoli cells by FSH and by other agents that increase cAMP levels. The cAMP effect is exerted at the transcriptional level and appears to be cell type specific, since it is not observed in other KL-expressing primary cells or cell lines. Deletion analysis of the 5'-flanking region of the mouse KL gene shows that the proximal promoter sequence between -88 and +8 from the transcriptional start site is necessary and sufficient to obtain the full cAMP responsiveness of the promoter in primary mouse Sertoli cells. In the -88/+8 promoter region, several cis-acting elements play a role in cAMP response. The -88/-56 sequence is necessary for full induction of the gene, since its removal causes a drastic decrease in cAMP responsiveness; however, cAMP-stimulated expression is still observed with the minimal promoter region between -55 and +8. A more detailed mutational analysis of the minimal promoter region shows that mutations in the canonical TATA box sequence and in an immediately downstream GC-rich element completely abolish cAMP responsiveness. DNA-binding experiments show that transcription factor Sp1 binds to the -88/-56 fragment of the KL proximal promoter in both control and cAMP stimulated cells, whereas a new cAMP-induced complex is observed when the -55/+8 minimal promoter region is used as probe. The canonical TATA box sequence is essential for formation of the latter complex. We also show that the binding of an unknown nuclear factor (different from Sp1, Egr-1, Rnf6, and AP-2) to a GC rich element between -19 and +8 increases after cAMP treatment, and this effect seems to be specific of primary Sertoli cells. Thus, cAMP-induced transcription from the KL gene promoter in primary mouse Sertoli cells is mediated by a complex interaction among a Sp1-binding region, factors recognizing the canonical TATA box sequence, and a not yet identified cAMP-induced factor binding a GC-rich sequence just downstream from it. PMID- 12904319 TI - Bacterially-induced preterm labor and regulation of prostaglandin-metabolizing enzyme expression in mice: the role of toll-like receptor 4. AB - Toll-like receptor 4 (TLR-4) is a critical mediator of the cellular response to lipopolysaccharide. Our purpose was to examine the role of TLR-4 in parturition and in the regulation of expression of prostaglandin synthase (cyclooxygenase [COX]-1 and COX-2) and 15-hydroxyprostaglandin dehydrogenase (PGDH) following exposure to heat-killed Escherichia coli (HKE) in pregnant mice. Inbred TLR-4 mutant C3H/HeJ mice and inbred normal C3HeB/FeJ mice on Day 14.5 of a 19- to 20 day gestation received intrauterine injection of either HKE or sterile vehicle (PBS). Preterm or term delivery was recorded for these animals. Tissues (myometrium, decidual caps, placentas, fetal membranes, and fetuses) were collected after injection of sterile vehicle or 5 x 109 HKE bacteria (n = 5 mice per strain per treatment per time point). The COX-1, COX-2, and PGDH gene expression was determined by semiquantitative reverse transcription-polymerase chain reaction. We found that 5 x 109 HKE induced preterm delivery in 100% of TLR 4-normal mice but in 0% of TLR-4-mutant mice. The HKE exposure up-regulated expression of COX-2, but not of COX-1, in maternal tissues in both mouse strains. The prostaglandin-catabolizing enzyme PGDH was down-regulated in myometrium, fetal membranes, and fetuses in control mice, but no change was observed in TLR-4 mutant mice after HKE treatment. These results demonstrate that a functional TLR 4 is essential for HKE-induced preterm labor and PGDH down-regulation but is not essential for HKE-induced COX-2 gene up-regulation. The TLR-4 may mediate bacterially induced preterm labor via regulation of prostaglandin degradation rather than prostaglandin synthesis. PMID- 12904321 TI - Fetal lungs of tenascin-C-deficient mice grow well, but branch poorly in organ culture. AB - Tenascin-C (TNC) is a multidomain extracellular matrix protein that contributes to organogenesis and tumorgenesis. To elucidate its developmental function in the context of TNC deficiency, lung lobes of TNC null mice were obtained at Embryonic Days E11.5 and E12.5 and cultured for 3 d. In lung explants of homozygote TNC deficient embryos (E12.5) the number of future airway branches was reduced by 36% as compared with wild-type. In heterozygote explants only half of the reduction (18%) was observed. No significant alteration, neither of the explant growth nor of the pattern of airway branching, was noticed in TNC-null explants. However, the terminal endbuds of the transgenic explants were enlarged. The results are supported by a morphologic investigation at Postnatal Day P2, where the airspaces of TNC-deficient lungs appeared larger than in wild-type lungs. Taken together, our results represent the first developmental phenotype of TNC-null mice. We conclude that TNC takes part in the control of fetal lung branching, and that not only the presence of TNC but also its amount is important. Because TNC is predominantly expressed at the growing tip of the future airways, we hypothesize that TNC promotes the penetration into the surrounding mesenchyme and the branching of the growing airways. PMID- 12904320 TI - Tolerance of the mouse sperm nuclei to freeze-drying depends on their disulfide status. AB - Mouse spermatozoa from the caudae epididymides could be freeze-dried without losing their ability to support normal development. Immature spermatozoa from the testes, in contrast, were damaged by freeze-drying. However, immature spermatozoa became resistant to freeze-drying after their treatment with diamide, which oxidizes free -SH groups. Conversely, epididymal spermatozoa were damaged by freeze-drying if first treated with dithiothreitol (DTT), which reduces -SS- bonds. The potential for freeze-drying damage seems likely to relate to the -SS- status of sperm proteins, in particular its protamines. PMID- 12904322 TI - Endoscopic ultrasound, positron emission tomography, and computerized tomography for lung cancer. AB - Staging of patients with lung cancer to determine operability is intended to efficiently limit futile thoracotomies without denying possibly curative surgery. Currently available staging tests are imperfect alone and in combination. Imaged suspected metastases often require tissue confirmation before surgery can be denied. Endoscopic ultrasound (EUS) may help identify inoperable patients by providing tissue proof of inoperability in a single staging test, with similar sensitivity for identifying inoperable patients as other staging tests. Therefore, we compared computed tomography, positron emission tomography (PET), and EUS with fine-needle aspiration under conscious sedation, each test interpreted blinded with respect to the other tests, for identifying inoperable patients in a consecutive cohort of 79 potentially operable patients with suspected or proven lung cancer. An economic analysis was also performed. Thirty nine patients were found inoperable (a 40th patient's inoperability was missed by all preoperative staging tests). The sensitivity of computerized tomography was 43%. PET and EUS each had similar sensitivities (68 and 63%, respectively) and similar negative predictive values (64 and 68%, respectively), but EUS's superior specificity (100 vs. 72% for PET) and considerably lower expense means it may be preferred to PET early in staging to identify inoperable patients. PMID- 12904323 TI - Bronchoscopic microsampling method for measuring drug concentration in epithelial lining fluid. AB - Direct measurement of the concentration of antimicrobial agents in bronchial epithelial lining fluid (ELF) would allow for a more informed approach to appropriate dosing of antimicrobial agents for respiratory tract infections. In this study, we determined the time versus concentration profile in ELF after an oral administration of levofloxacin, using recently developed bronchoscopic microsampling probes. These probes could be repeatedly and safely inserted through the fiberoptic bronchoscope in normal healthy volunteers. The concentration of levofloxacin in ELF was 43.4% of the corresponding serum value at 1 hour, reached the same level at 2 hours, decreased in a similar manner as that in serum, and returned to undetectable levels at 24 hours. It exceeded minimal inhibitory concentrations of Staphylococcus aureus (0.25 microg/ml), Klebsiella species (0.5 microg/ml), and Haemophilus influenzae (0.06 microg/ml) after 6 hours. The experimental procedure was well tolerated, and no complications were observed. In conclusion, bronchoscopic microsampling is a feasible and promising method for measuring antimicrobial concentrations in the target sites of respiratory tracts directly and repeatedly. PMID- 12904324 TI - Fish oil supplementation reduces severity of exercise-induced bronchoconstriction in elite athletes. AB - In elite athletes, exercise-induced bronchoconstriction (EIB) may respond to dietary modification, thereby reducing the need for pharmacologic treatment. Ten elite athletes with EIB and 10 elite athletes without EIB (control subjects) participated in a randomized, double-blind crossover study. Subjects entered the study on their normal diet, and then received either fish oil capsules containing 3.2 g eicosapentaenoic acid and 2.2 g docohexaenoic acid (n-3 polyunsaturated fatty acid [PUFA] diet; n = 5) or placebo capsules containing olive oil (placebo diet; n = 5) taken daily for 3 weeks. Diet had no effect on preexercise pulmonary function in either group or on postexercise pulmonary function in control subjects. However, in subjects with EIB, the n-3 PUFA diet improved postexercise pulmonary function compared with the normal and placebo diets. FEV1 decreased by 3 +/- 2% on n-3 PUFA diet, 14.5 +/- 5% on placebo diet, and 17.3 +/- 6% on normal diet at 15 minutes postexercise. Leukotriene (LT)E4, 9alpha, 11beta-prostaglandin F2, LTB4, tumor necrosis factor-alpha, and interleukin-1beta, all significantly decreased on the n-3 PUFA diet compared with normal and placebo diets and after the exercise challenge. These data suggest that dietary fish oil supplementation has a markedly protective effect in suppressing EIB in elite athletes, and this may be attributed to their antiinflammatory properties. PMID- 12904325 TI - Progressive decline in plethysmographic lung volumes in infants: physiology or technology? AB - During the last 30 years, there has been an unexplained trend toward declining values for plethysmographic assessments of lung volume at functional residual capacity (FRC) in infants. The aim of this study was to compare data collected from healthy infants using contemporary equipment with published reference data and to explore reasons for discrepancies. Lung volumes were measured in 32 healthy infants (age, 4-93 weeks; weight, 3.9-12.4 kg) using a new, commercially available infant plethysmograph. Mean (SD) FRC was 19.6 (3.4) ml/kg (within subject coefficient of variation 3.4 [2.3%]), which was on average 7.0 [3.5] ml/kg and 2.3 [1.2] SD (Z) scores lower than the recently collated reference data from an American Thoracic Society task force. A total of 66% of these healthy infants had a FRC that was below the predicted normal range. Comparison of equipment, software, and protocols with those from previous reports revealed the importance of minimization of dead space and of adequate subtraction of all compressible occluded volume when calculating FRC in infants. These findings emphasize the need to establish reference data for lung function tests in infants that are appropriate for the equipment and protocols in current use. PMID- 12904327 TI - Obstructive sleep apnea and thoracic aorta dissection. AB - Obstructive sleep apnea syndrome (OSAS) is a process that is associated with the development of arterial hypertension, the main risk factor for aortic dissection and during obstructive episodes of the upper airways with marked increases in transmural pressure of the aorta wall. The aim of this work was to study the association between aortic dissection and OSAS. Nineteen consecutive patients with thoracic aorta dissection and 19 hypertensive patients of similar age, sex, and body mass index were studied by clinical questionnaire and polysomnography. Snoring and nonrefreshing sleep were common in both groups. Thirteen patients (68%) from each group showed an apnea-hypopnea index of more than 5 per hour. However, patients with aortic dissection presented a higher apnea-hypopnea index (28 [30.3] versus 11.1 [10.4], p=0.032). Seven patients with dissection presented an apnea-hypopnea index of more than 30 versus 1 patient in the control group (p=0.042). Patients with thoracic aorta dissection presented a high prevalence of previously undiagnosed and frequently severe OSAS. Further studies, including this diagnosis as a prognostic variable in the follow-up of patients with aortic dissection, are required. Our results suggest that in patients with aortic dissection and symptoms consistent with OSAS, a sleep study should be considered in their clinical management. PMID- 12904326 TI - Surfactant protein profile of pulmonary surfactant in premature infants. AB - Although premature infants are known to be deficient in pulmonary surfactant, there is limited information regarding surfactant protein (SP) composition. To assess the postnatal profile of SPs, tracheal aspirate samples were collected from 35 intubated infants of 23-31 weeks of gestation between 8 and 80 days of age. In 71 large aggregate surfactant samples that had normal in vitro function (minimum surface tension of less than 1 mN/m by pulsating bubble surfactometry), mean +/- SEM contents of SP-A, SP-B, and SP-C (3.7 kD) were 7.1 +/- 1.4%, 1.8 +/- 0.2%, and 4.6 +/- 0.6%, respectively, of phospholipid. To assess SPs in the 1st week of life, we analyzed samples from additional infants receiving only synthetic replacement surfactant. On the 2nd day of life, contents of SP-A, SP-B, and SP-C were 13.4%, 8.4%, and 0.1%, respectively, of the mean levels for Day 8 80 samples. The major postnatal increases for SP-A, SP-B, and SP-C occurred during the 1st, 2nd, and 3rd weeks, respectively. We conclude that surfactant of newborn premature infants is markedly deficient in SPs, in particular SP-C. Despite continuing lung disease, some infants who are more than 1 week of age have surfactant with normal in vitro function that contains SPs at levels comparable to adult surfactant. PMID- 12904330 TI - How are complex cell properties adapted to the statistics of natural stimuli? AB - Sensory areas should be adapted to the properties of their natural stimuli. What are the underlying rules that match the properties of complex cells in primary visual cortex to their natural stimuli? To address this issue, we sampled movies from a camera carried by a freely moving cat, capturing the dynamics of image motion as the animal explores an outdoor environment. We use these movie sequences as input to simulated neurons. Following the intuition that many meaningful high-level variables, e.g., identities of visible objects, do not change rapidly in natural visual stimuli, we adapt the neurons to exhibit firing rates that are stable over time. We find that simulated neurons, which have optimally stable activity, display many properties that are observed for cortical complex cells. Their response is invariant with respect to stimulus translation and reversal of contrast polarity. Furthermore, spatial frequency selectivity and the aspect ratio of the receptive field quantitatively match the experimentally observed characteristics of complex cells. Hence, the population of complex cells in the primary visual cortex can be described as forming an optimally stable representation of natural stimuli. PMID- 12904328 TI - Increased renal ENaC subunit and sodium transporter abundances in streptozotocin induced type 1 diabetes. AB - Uncontrolled diabetes mellitus (DM) is associated with copious water and sodium losses. We hypothesized that the kidney compensates for these losses by increasing the abundances of key sodium and water transporters and channels. Using targeted proteomic analysis via immunoblotting of kidney homogenates, we examined comprehensive regulation of transport proteins. In three studies, streptozotocin (STZ; 65 mg/kg) or vehicle was administered intraperitoneally to male Sprague-Dawley rats. In study 2, to control for potential renal toxicity of STZ, one group of STZ-treated rats was intensively treated with insulin to control diabetes. In another group, the reversibility of DM and related changes was assessed by treating animals with insulin for the final 4 days. In study 3, we correlated blood glucose to transporter changes by treating animals with different doses of insulin. In study 1, STZ treatment resulted in significantly increased band densities for the type 3 sodium/hydrogen exchanger (NHE3), the thiazide-sensitive Na-Cl cotransporter (NCC), and epithelial sodium channel (ENaC) subunits alpha, beta, and gamma (85- and 70-kDa bands) to 204, 125, 176, 132, 147, and 241% of vehicle mean, respectively. In study 2, aquaporin-2 (AQP2) and AQP3 were increased with DM, but not AQP1 or AQP4. Neither these changes, nor blood glucose itself, could be returned to normal by short-term intensive insulin treatment. Whole kidney abundance of AQP3, the bumetanide-sensitive Na-K-2Cl cotransporter (NKCC2), and gamma-ENaC (85-kDa band) correlated most strongly with blood glucose in study 3. These comprehensive changes would be expected to decrease volume contraction accompanying large-solute and water losses associated with DM. PMID- 12904329 TI - Overexpression of PKC-betaI and -delta contributes to higher PKC activity in the proximal tubules of old Fischer 344 rats. AB - Previously, we reported that natriuretic and diuretic response to dopamine is diminished in old Fischer 344 rats, which is due to higher basal protein kinase C (PKC) activity and hyperphosphorylation of Na-K-ATPase in the proximal tubules (PTs) of old rats. The present study was conducted to determine whether higher PKC activity could be due to altered expression of some of the PKC isoforms in the superficial cortex (rich in PTs) of old rats. Fluorimetric measurement showed almost twofold increase in the PKC activities in homogenates and membranes of old (24 mo) compared with adult (6 mo) rats. Interestingly, in the basal state PKC betaI was overexpressed in the membranes, whereas PKC-delta expression was increased in the cytosol of old compared with adult rats. Treatment of the cortical slices with either SKF-38393, a D1-like agonist, or PDBu, a direct activator of PKC, caused translocation of PKC-betaI from cytosol to membranes in adult but not in old rats. Both of these drugs caused translocation of PKC-delta from membranes to cytosol in adult but not in old rats. These drugs had no effect on translocation of PKC-zeta in both adult and old rats. Both PKC-betaI and delta co-immunoprecipitated with alpha1-subunit of Na-K-ATPase in adult and old rats. These observations suggest that both SKF-38393 and PDBu differentially regulate PKC-betaI and -delta in adult but not in old rats. Also, PKC-betaI and delta seem to interact with Na-K-ATPase in these animals. The overexpression of both PKC-betaI and -delta in old rats could be responsible for a higher basal PKC activity, which causes the hyperphosphorylation of Na-K-ATPase and contributes to the diminished inhibition of Na-K-ATPase activity by dopamine in old rats. PMID- 12904332 TI - Strategies for the integration of posture and movement during reaching in the cat. AB - We have examined the relationship between the movement and the anticipatory postural adjustments (APAs) that precede that movement during a reaching task in the cat. We recorded ground reaction forces in all 3 planes from all 4 limbs as well as electromyographic (EMG) activity from limb and axial muscles. The reaching movement was always preceded by an APA that was characterized by a loading of the reaching forelimb and an unloading of the support forelimb. This loading of the reaching forelimb was preceded, and accompanied, by increased activity in shoulder and limb extensor muscles of the reaching limb; extensor muscle activity in the supporting limb was simultaneously decreased. An important finding from this study was that the onset of the APA and of the movement was temporally decoupled. Analyses of the onset of EMG activity showed that most of the muscles that we recorded could be classified as either related to the APA or related to the movement. These results support the idea of distributed, and perhaps independent, systems for the execution of the APA and of the prime movement. There was also postural activity in the supporting limb during the movement. Analysis of this activity, which is also anticipatory in nature, suggests that it was tightly linked to the movement. We suggest that this postural response is signaled as part of the command for movement. Some muscles, particularly the extensors of the reaching limb, received convergent input from the command signals for the APA and for the movement. PMID- 12904331 TI - PI3-kinase inhibitors abolish the enhanced chronotropic effects of angiotensin II in spontaneously hypertensive rat brain neurons. AB - Angiotensin II (Ang II), acting at Ang II type 1 receptors (AT1Rs), increases the firing rate of neurons from Wistar-Kyoto (WKY) rat brain via protein kinase C (PKC)- and calcium-calmodulin kinase II (CaMKII)-dependent mechanisms. The objectives of this study were twofold; first, to compare the Ang-II-stimulated increase in firing of neurons from WKY and spontaneous hypertensive rats (SHR) and second, to elucidate the signaling mechanisms involved. Action potentials were measured in neurons cultured from SHR and WKY rat brains using the whole cell configuration of the patch-clamp technique in the current-clamp mode. Ang II (100 nM) caused three- and sixfold increases in neuronal firing rate in WKY rat and SHR neurons, respectively; effects that were abolished by the AT1R antagonist Losartan (1 microM). Co-administration of calphostin C (10 microM, a PKC inhibitor) and KN-93 (10 microM, a CaMKII inhibitor) completely blocked this Ang II action in WKY rat neurons, while they caused only a approximately 50% attenuation in SHR neurons. The residual increase in firing rate produced by Ang II in SHR neurons was blocked by inhibitors of phosphatidylinositol 3 kinase (PI3 kinase), either LY 294002 (10 microM) or wortmannin (100 nM). These observations suggest that a PI3-kinase signaling pathway may be responsible for the enhanced chronotropic effect produced by Ang II in SHR neurons. PMID- 12904333 TI - Contrasting neuronal activity in the supplementary and frontal eye fields during temporal organization of multiple saccades. AB - The organization of a series of actions into an appropriate temporal order is of particular importance in the voluntary control of motor behavior. Previous reports have emphasized the importance of medial motor areas for the temporal organization of movements. The aim of this study was to compare the neuronal activity in the supplementary and frontal eye fields (SEF and FEF) during sequential performance of multiple saccades to clarify the role of the two cortical oculomotor areas in the temporal organization of saccades based on memorized information. We analyzed neuronal activity while monkeys performed three saccades to peripheral targets in orders that were instructed and memorized. We found that activity that reflected saccade sequence or the numerical position of a saccade within a sequence (rank) was more prevalent in the SEF, whereas activity reflecting saccade direction was more dominant in the FEF. Furthermore, a sizeable number of SEF neurons exhibited an increase in activity when the animals were required to discard a current sequence and compose a novel sequence. We propose that the SEF is primarily involved in the process of planning, decoding, and updating saccade sequences, whereas the FEF plays a major role in determining the direction of forthcoming saccades. PMID- 12904334 TI - Phenytoin protects spinal cord axons and preserves axonal conduction and neurological function in a model of neuroinflammation in vivo. AB - Axonal degeneration within the spinal cord contributes substantially to neurological disability in multiple sclerosis (MS). Thus neuroprotective therapies that preserve axons, so that they maintain their integrity and continue to function, might be expected to result in improved neurological outcome. Sodium channels are known to provide a route for sodium influx that can drive calcium influx, via reverse operation of the Na+/Ca2+ exchanger, after injury to axons within the CNS, and sodium channel blockers have been shown to protect CNS axons from degeneration after experimental anoxic, traumatic, and nitric oxide (NO) induced injury. In this study, we asked whether phenytoin, which is known to block sodium channels, can protect spinal cord axons from degeneration in mice with experimental allergic encephalomyelitis (EAE), which display substantial axonal degeneration and clinical paralysis. We demonstrate that the loss of dorsal corticospinal tract (63%) and dorsal column (cuneate fasciculus; 43%) axons in EAE is significantly ameliorated (corticospinal tract: 28%; cuneate fasciculus: 17%) by treatment with phenytoin. Spinal cord compound action potentials (CAP) were significantly attenuated in untreated EAE, whereas spinal cords from phenytoin-treated EAE had robust CAPs, similar to those from phenytoin treated control mice. Clinical scores in phenytoin-treated EAE at 28 days were significantly improved (1.5, i.e., minor righting reflex abnormalities) compared with untreated EAE (3.8, i.e., near-complete hindlimb paralysis). Our results demonstrate that phenytoin has a protective effect in vivo on spinal cord axons, preventing their degeneration, maintaining their ability to conduct action potentials, and improving clinical status in a model of neuroinflammation. PMID- 12904335 TI - KCC2 mediates NH4+ uptake in cultured rat brain neurons. AB - Elevated levels of NH4+ in the brain impair neuronal function. We studied the effects of NH4+ on postsynaptic inhibition of cultured rat brain neurons using whole cell recording under nominally HCO3- -free conditions. Application of NH4+ shifted the reversal potentials for spontaneous inhibitory postsynaptic currents and currents elicited by dendritic GABA applications in a positive direction because [Cl-]i increased. The positive shift of the reversal potentials of GABA induced Cl- currents was equal on equimolar elevation of [NH4+]o or [K+]o, respectively. The NH4+-induced increase in [Cl-]i was reversed by an inhibitor of cation-anion cotransport, furosemide (0.1 mM), but not by bumetanide (0.01 mM) or by replacement of [Na+]o by Li+. We conclude that neuron-specific K-Cl cotransporter (KCC2) transports NH4+ similar to K+. Despite this fact, the small increase of [NH4+]o during metabolic encephalopathies will barely elevate [Cl-]i. However, an impairment of neuronal function may result because KCC2 provides a pathway to accumulate NH4+, and thereby, a continuous acid load to neurons. PMID- 12904336 TI - Vibrissa movement elicited by rhythmic electrical microstimulation to motor cortex in the aroused rat mimics exploratory whisking. AB - The rhythmic motor activity of the vibrissae that rodents use for the tactile localization of objects provides a model system for understanding patterned motor activity in mammals. Evidence suggests that neural circuitry in the brain stem provides rhythmic drive to the vibrissae. Yet multiple brain structures at higher levels of organization, including vibrissa primary motor cortex (M1), have direct projections to brain stem nuclei that are implicated in whisking. We thus asked whether output from M1 can control vibrissa movement on the approximately 10-Hz scale of the natural rhythmic movement of the vibrissae. Our assay of cortical control made use of periodic intracortical microstimulation (ICMS) to excite a region of vibrissa M1 cortex in awake, behaving animals and measurements of the stimulus-locked electromyogram (EMG) in both the intrinsic and extrinsic muscles that drive the vibrissae. We observed that ICMS evoked a prompt activation of the extrinsic muscles and a delayed and prolonged response in the intrinsic muscles. The relative timing and shape of these waveforms approximates the EMG waveforms seen during natural exploratory whisking. We further observed prompt activation of the intrinsic muscles, an occurrence not seen during exploratory whisking. Despite the latter difference in muscular activation, the motion of the vibrissae evoked by periodic ICMS strongly resembled the motion during natural, exploratory whisking. Interestingly, the extent of the movement was proportional to the level of arousal, as quantified by the amplitude of hippocampal activity in the theta frequency band. We interpret these data as demonstrating that M1 cortex can, in principle, initiate the full pattern of whisking on a cycle-by-cycle basis in aroused animals. Beyond issues of natural motor control, our result may bear on the design of algorithms for neuroprosthetic control of motor output. PMID- 12904337 TI - Loss of alpha CGRP reduces sound-evoked activity in the cochlear nerve. AB - alpha-Calcitonin gene-related peptide (alphaCGRP) is one of several neurotransmitters immunolocalized in the unmyelinated component of the cochlear efferent innervation, the lateral olivocochlear (OC) system, which makes axo dendritic synapses with cochlear sensory neurons. In rodents, CGRP is also immunocolocalized in the myelinated medial OC system, which contacts cochlear outer hair cells (OHCs). To understand the role(s) of this neuropeptide in the OC system, we characterized the auditory phenotype of alphaCGRP-null mice. Cochlear threshold sensitivity was normal in mutant mice, both via a neural metric, the auditory brain stem response (ABR), and an OHC-based metric, distortion product otoacoustic emissions (DPOAEs). Medial OC function and resistance to acoustic injury were also unaffected by alphaCGRP deletion: the former was assessed by measuring cochlear response suppression with electrical stimulation of the OC bundle, the latter by measuring temporary threshold shifts after exposure to high level sound. However, significant abnormality in alphaCGRP-null mice was seen in the growth of cochlear neural responses with increasing stimulus level. This observation, contrasted with normal amplitude-versus-level functions for DPOAEs, is consistent with a selective, postsynaptic effect on cochlear neurons via alphaCGRP release from lateral OC terminals. This constitutes the most direct evidence to date for a functional role of the lateral OC system in the auditory periphery. PMID- 12904338 TI - Absence of local sign withdrawal in chronic human spinal cord injury. AB - Local sign withdrawal, a reflex to direct the limb away from noxious cutaneous stimuli, is thought to be indicative of a modular organization of the spinal cord. To assess the integrity of such an organization of the spinal cord in chronic human spinal cord injury (SCI), we tested the electromyogram (EMG) and joint torque responses to cutaneous stimuli applied to 6 locations of the leg in 10 SCI volunteers and 3 spinal-intact controls. The 6 locations included the medial arch of the foot, the second metatarsal, the dorsum, the region over the sural nerve at the lateral malleolus, and the anterior and posterior aspects of the lower leg. Although spinal-intact subjects demonstrated local sign withdrawal, the data from SCI subjects indicated that an invariant flexion response pattern was produced regardless of stimulus location. Ankle dorsiflexion and hip flexion were produced in all subjects at all locations and no difference in the ratio of hip:ankle torques could be detected for the 6 test locations. A windup-crossover test, employing a sequence of 6 stimuli at 1-s intervals was used to assess whether common neuronal pathways were responsible for the loss of modular organization. An additional 10 SCI volunteers were tested using stimuli in which the stimulus location was switched between the 2nd and 3rd stimulus of the test sequence. The response to the crossover stimulus more closely resembled the response to the 3rd stimulus of a windup sequence than a response without conditioning stimuli. These results indicate that increased excitability produced by windup at one stimulus site is maintained at the 2nd site. This observation suggests that deep dorsal horn neurons, typically associated with musculotopic mapping, may be reorganized in chronic spinal cord injury. PMID- 12904339 TI - Acute modulation of adrenal chromaffin cell BK channel gating and cell excitability by glucocorticoids. AB - Although adrenal glucocorticoids cortisol and corticosterone (CORT) have numerous "genomic" effects on adrenomedullary chromaffin cells, acute modulatory actions remain largely unknown, despite rapid stress-related changes in secretion. We report that 1 microM glucocorticoids rapidly modulate gating of chromaffin cell BK channels and action potential firing. In general, CORT, or the analog dexamethasone (DEX), increased channel activity in inside-out bovine patches, an effect not blocked by the glucocorticoid receptor (GR) antagonist RU38486. By contrast, these steroids could profoundly inhibit BK activation in many rat patches, while facilitating activation in others. We show that BK inhibition arises from a negative shift in the voltage dependence of BK inactivation paralleling that for activation. We report that rat cells characteristically exhibit greater repetitive firing ability than bovine cells in the absence of glucocorticoids. In both species, steroid application typically increased firing responses to smaller current injections, attributable to BK-enhanced repolarization and Na+ channel deinactivation. However, in rat cells, where BK inactivation is generally faster and more complete, glucocorticoids tended to dampen responses to stronger stimuli. Thus, in the context of natural variation in BK gating, glucocorticoids can either promote or limit firing responses. We suggest that steroids exploit BK gating variety to tailor catecholamine output in a species- and context-specific fashion. PMID- 12904340 TI - Role of the microbiology laboratory in diagnosis and management of pharyngitis. PMID- 12904341 TI - Typing of nonencapsulated haemophilus strains by repetitive-element sequence based PCR using intergenic dyad sequences. AB - Intergenic dyad sequences (IDS) are short repeated elements that have been described for several Haemophilus genomes and for only two other bacterial genera. We developed a repetitive-element sequence-based PCR using an IDS specific primer as a typing method (IDS-PCR) for nonencapsulated Haemophilus strains and compared this technique with pulsed-field gel electrophoresis (PFGE) of DNA restricted with SmaI. IDS-PCR was rapid, easy to perform, and reproducible, with a high discriminatory capacity for nontypeable Haemophilus influenzae (NTHI) strains. The 69 NTHI strains tested generated 65 different banding patterns. Epidemiologically related strains gave similar or identical fingerprints, and all of the unrelated strains except two showed different patterns. These results were in agreement with those obtained by PFGE. For 20 genital strains usually identified as being biotype IV NTHI and belonging to a cryptic genospecies of Haemophilus with remarkable genetic homogeneity, four bands were significantly present and six bands were significantly absent from the fingerprints. The 20 strains were gathered in 11 closely related profiles, whereas PFGE provided no band when DNA was treated with SmaI. IDS-PCR improved the differentiation previously obtained within this species by ribotyping and multilocus enzyme electrophoresis. Our findings suggest that IDS-PCR is a rapid, reliable, and discriminatory method for typing NTHI strains and is currently the most efficient method for distinguishing strains within the cryptic genospecies of HAEMOPHILUS: PMID- 12904342 TI - Identification of Streptococcus sanguinis with a PCR-generated species-specific DNA probe. AB - The objective of the present study was to design a PCR-generated DNA probe and determine the specificity of the probe for the identification of clinical isolates of Streptococcus sanguinis. To do this, we examined over 200 arbitrarily primed PCR (AP-PCR) amplicon patterns obtained with DNA from clinical isolates of S. sanguinis. A 1.6-kb DNA amplicon that was common to all AP-PCR profiles was extracted from agarose gels and then cloned and sequenced. A search for a similar sequence in the GenBank database with the BLASTN program revealed that the 1.6-kb DNA fragment comprised an intergenic region between two housekeeping genes, uncC (proton-translocating ATPase) and murA (UDP-N-acetylglucosamine enolpyruvyl transferase). Three digoxigenin-labeled DNA probes were synthesized on the basis of the sequence of the 1.6-kb fragment: the sequence of probe SSA-1 contained the proton-translocating ATPase (uncC) and the entire intergenic region, the sequence of probe SSA-2 contained only the intergenic region, and the sequence of probe SSA-3 contained an internal region of the murA gene. Dot blot hybridization showed that the three probes displayed signals for hybridization to both S. sanguinis strain ATCC 10556 and the S. sanguinis clinical isolates. Probe SSA-1, however, hybridized to DNA from S. oralis and S. mitis. Probe SSA-3 hybridized to DNA from S. gordonii, S. mitis, S. oralis, S. parasanguinis, and S. vestibularis. The probe SSA-2-specific intergenic region appeared to be specific for S. sanguinis. The results from this study suggest that probe SSA-2 may serve as a species-specific DNA probe for the identification of clinical isolates of S. sanguinis. PMID- 12904343 TI - Comparison of the Directigen flu A+B test, the QuickVue influenza test, and clinical case definition to viral culture and reverse transcription-PCR for rapid diagnosis of influenza virus infection. AB - The diagnostic performances of the clinical case definition of influenza virus infection based on the combination of fever and cough and of two rapid influenza diagnostic tests, the Directigen Flu A+B test (Directigen; BD Diagnostic Systems, Sparks, Md.) and the QuickVue influenza test (QuickVue; Quidel, San Diego, Calif.), were compared to those of viral culture and an in-house reverse transcription (RT)-PCR during the 2000-2001 flu season. Two hundred consecutive nasopharyngeal aspirates were analyzed from 192 patients, including 122 adults and 70 children. Viral culture identified influenza virus A in 16 samples and influenza virus B in 55 samples, whereas RT-PCR identified influenza virus A in 21 samples and influenza virus B in 64 samples. When RT-PCR was used as the reference standard, the likelihood ratios for a positive test were 40.0 for Directigen, 8.6 for QuickVue, and 1.4 for the combination of fever and cough, whereas the likelihood ratios for a negative test were 0.22, 0.16, and 0.48, respectively. Our study suggests that (i). the poor specificity (35 to 58%) and the poor positive predictive value (41 to 60%) of the clinical case definition of influenza preclude its use for prediction of influenza virus infections during epidemics, especially when infection control decision making in the hospital setting is considered; (ii). Directigen has a higher diagnostic yield than QuickVue but is associated with a larger number of invalid results; (iii). the sensitivities of the rapid diagnostic tests are significantly lower with samples from adults than with samples from children, with the rates of false-negative results reaching up to 29%; and (iv). RT-PCR detects more cases of influenza than viral culture, and this greater accuracy makes it a more useful reference standard. PMID- 12904344 TI - Epidemiological survey of Babesia species in Japan performed with specimens from ticks collected from dogs and detection of new Babesia DNA closely related to Babesia odocoilei and Babesia divergens DNA. AB - Detection and analysis of Babesia species from ticks recovered from dogs in Japan were attempted by PCR and nucleotide sequence analysis based on the 18S rRNA gene, respectively. A total of 1136 ticks were examined for Babesia DNA by 18S rRNA-based PCR and nucleotide sequencing. Partial sequences of Babesia canis vogeli DNA were detected from six ticks in Aomori, Nara, Hiroshima, Oita, and Okinawa Prefectures; and Babesia gibsoni Asia-1 DNA was also detected in four ticks in Osaka, Hiroshima, Miyazaki, and Okinawa Prefectures. Unique sequences of 1678 bp were also obtained from Ixodes ovatus ticks in Akita and Fukui Prefectures. The sequences were similar to those of Babesia odocoilei (97.7%) and Babesia divergens (97.6%). This is the first report of the detection of DNA belonging to this group in Japan. PMID- 12904345 TI - Evaluation of a novel medium for screening specimens from hospitalized patients to detect methicillin-resistant Staphylococcus aureus. AB - A novel medium, Oxacillin Resistant Screening Agar (ORSA) medium, was evaluated for the screening of specimens for methicillin-resistant Staphylococcus aureus (MRSA) in the hospital setting. Screening swabs (swabs of the nose, throat, perineum, and infected sites) were inoculated onto the new ORSA medium and into an enrichment broth (Muller-Hinton broth supplemented with NaCl and oxacillin). After 24 h of incubation, the enrichment broth was subcultured onto one ORSA plate and one lipovitellin Chapman salt agar plate. The sensitivities for the detection of MRSA were calculated for each medium alone and for the media in combination. A low sensitivity (74%) was obtained when ORSA medium was used alone as a primary culture, whereas the sensitivity was 88% when a single selective enrichment broth was used. Among the 414 blue colonies observed on ORSA plates, only 47% were found to be MRSA, 40% were coagulase-negative staphylococci, 7% were Enterococcus species, and 2% were methicillin-sensitive S. aureus. The optimal incubation time for the ORSA plates was evaluated. On primary culture, 38% of the blue MRSA colonies were visible only after 48 h of incubation (no blue colonies were not seen after 24 h of incubation), whereas 94% of the colonies were already visible at 24 h when ORSA plates were used for subcultures. In conclusion, the advantage of the novel ORSA medium is the ease of recognition of mannitol-fermenting bacteria, but further identification tests are needed to confirm the identification of S. aureus. An enrichment broth is still needed to ensure a good sensitivity for the recovery of MRSA, and an incubation time of 48 h is required for primary culture on ORSA medium. PMID- 12904346 TI - Evaluation of the MagNA pure LC instrument for extraction of hepatitis C virus RNA for the COBAS AMPLICOR Hepatitis C Virus Test, version 2.0. AB - The COBAS AMPLICOR system has played a major role in the transition of molecular diagnostics from research to routine clinical laboratory use by automating the nucleic acid amplification and detection processes. However, sample preparation remains a labor-intensive portion of the procedure. In this study, we evaluated the performance of the COBAS AMPLICOR Hepatitis C Virus Test, version 2.0 (Roche Molecular Systems, Branchburg, N.J.) following manual hepatitis C virus (HCV) RNA extraction versus automated extraction with the MagNA Pure LC instrument (Roche Applied Science, Indianapolis, Ind.). Parallel replicate testing was performed with standard dilutions of 100, 75, 60, and 0 HCV IU/ml and 153 clinical specimens. An analytical sensitivity of 75 IU/ml was achieved with either the manual or the standard-volume (200 microl) automated extraction methodologies (25 of 26 [96.2%]; 95% confidence interval [95% CI], 80.4 to 99.9), whereas the clinical sensitivity and specificity were both 100% with either extraction method. A large-volume (1 ml) automated extraction method was also evaluated with standard dilutions of 40, 25, 10, and 0 IU/ml and the same 153 clinical specimens. The analytical sensitivity of the COBAS AMPLICOR assay with the large volume extraction method was 25 HCV IU/ml (26 of 26 [100%]; 95% CI, 86.8 to 100), whereas the clinical sensitivity and specificity were both 100%. The MagNA Pure LC instrument is a versatile, labor-saving platform capable of integration with minimal modification of the existing assay procedure. The increased sensitivity of the COBAS AMPLICOR Hepatitis C Virus Test, version 2.0 performed in conjunction with large-volume HCV RNA extraction may be important in HCV diagnostic testing as new therapeutic strategies evolve. PMID- 12904347 TI - Comparison of molecular tests for detection and quantification of cell-associated cytomegalovirus DNA. AB - A cell-based standard was developed to compare the COBAS Amplicor CMV Monitor test, the Hybrid Capture System CMV DNA test, and the NucliSens CMV test. The standard was prepared by infecting human foreskin fibroblasts (HFFs) with cytomegalovirus (CMV) strain AD169 at low multiplicity of infection (0.03) and harvesting the cells at 6 h postinfection. Buffy coat cells were added to produce concentrations of from 0 to 10(5) HFFs per 10(6) total cells. Five laboratories performed the Amplicor PCR test and two laboratories performed the NucliSens and Hybrid Capture tests. The Amplicor PCR test was 1.5 to 2.0 log(10) more sensitive than the Hybrid Capture test. The specificities of the Amplicor PCR and Hybrid Capture tests were 100 and 93.8%, respectively. The linear range of the Amplicor PCR and Hybrid Capture tests were 2 to 4.48 log(10) and 3.48 to at least 5.0 log(10) CMV target cells, respectively. The standard deviations of the Amplicor PCR and Hybrid Capture tests varied throughout their linear range, and for both tests the variability was greater for lower concentrations of input CMV DNA. These data allow the direct comparison of viral load values between the Amplicor and Hybrid Capture tests. The analytical sensitivity of the NucliSens test could not be determined by using the 6-h standard, because the low multiplicity of infection and short culture time did not allow for adequate transcription of pp67 late mRNA measured in the test. Extending the incubation time of the standard to 24 h increased the analytical sensitivity of the NucliSens test to 3.0 log(10) target cells. PMID- 12904348 TI - Mycobacterial interspersed repetitive unit typing of Mycobacterium tuberculosis compared to IS6110-based restriction fragment length polymorphism analysis for investigation of apparently clustered cases of tuberculosis. AB - An evaluation of the utility of IS6110-based restriction fragment length polymorphism (RFLP) typing compared to a combination of variable number tandem repeat (VNTR) typing and mycobacterial interspersed repetitive unit (MIRU) typing was undertaken. A total of 53 patient isolates of Mycobacterium tuberculosis from four presumed episodes of cross-infection were examined. Genomic DNA was extracted from the isolates by a cetyl trimethylammonium bromide method. The number of copies of tandem repeats of the five loci ETR(A) to ETR(E) and 12 MIRU loci was determined by PCR amplification and agarose gel electrophoresis of the amplicons. VNTR typing identified the major clusters of strains in the three investigations in which they occurred (each representing a different evolutionary clade: 32333, 42235, and 32433). The majority of unrelated isolates (by epidemiology and RFLP typing) were also identified by VNTR typing. The concordance between the RFLP and MIRU typing was complete, with the exception of two isolates with RFLP patterns that differed by one band each from the rest of the major epidemiologically linked groups of isolates in investigation A. All of these isolates had identical MIRU and VNTR types. A further pair of isolates differed in the number of tandem repeat copies at two MIRU alleles but had identical RFLP patterns. The speed of the combined VNTR and MIRU typing approach enabled results for some of the investigations to be supplied in "real time," influencing choices in contact tracing. The ease of comparison of results of MIRU and VNTR typing, which are recorded as single multidigit numbers, was also found to greatly facilitate investigation management and the communication of results to health care professionals. PMID- 12904349 TI - Comparison of five genotypic techniques for identification of optochin-resistant pneumococcus-like isolates. AB - Three PCR techniques (amplification of the psaA, ply, and lytA genes) and a commercial kit (AccuProbe [GenProbe, San Diego, Calif.], based on hybridization with the 16S rRNA gene), all four of which claimed to be specific for Streptococcus pneumoniae, were used to identify 49 alpha-hemolytic streptococcal isolates suspected of being pneumococci. The definite phenotypic identification of these organisms as S. pneumoniae was difficult when optochin susceptibility and the presence of a capsule were taken as markers. Furthermore, RsaI digestion of the amplified 16S rRNA gene was applied. All 49 strains were optochin resistant. Eleven of these were encapsulated and were identified as pneumococci by all tests. Twenty of the 38 unencapsulated strains were unambiguously identified as nonpneumococci by all tests. The identities of another 18 unencapsulated strains remained inconclusive due to highly variable reactions for all phenotypic and genotypic techniques applied. The AccuProbe test was positive for seven strains for which the results of the other tests were inconclusive. RsaI restriction of the amplified 16S rRNA gene confirmed the AccuProbe result for all strains, while the result of the psaA-specific PCR was in concordance with encapsulation for all strains. The results presented here indicate that identification problems continue to exist for some strains, despite the application of genotypic and phenotypic tests in combination. We found the psaA specific PCR to be the genotypic technique best suited for the identification of genuine pneumococci and optochin-resistant pneumococci. PMID- 12904350 TI - Single-nucleotide-polymorphism mapping of the Pseudomonas aeruginosa type III secretion toxins for development of a diagnostic multiplex PCR system. AB - We mapped the coding single nucleotide polymorphisms in four toxin genes-exoS, exoT, exoU, and exoY-of the Pseudomonas aeruginosa type III secretion system among several clinical isolates. We then used this information to design a multiplex PCR assay based on the simultaneous amplification of fragments of these genes. Eight strains of known genotype were used to test our multiplex PCR method, which showed 100% sensitivity and specificity in this small sample size. This assay appears to be promising for the rapid and accurate genotyping of the presence of these genes in clinical strains of P. aeruginosa. PMID- 12904351 TI - Comparison of automated and manual nucleic acid extraction methods for detection of enterovirus RNA. AB - Automated nucleic acid extraction is an attractive alternative to labor-intensive manual methods. We compared two automated methods, the BioRobot M48 instrument (Qiagen, Inc.) and MagNA Pure (Roche Applied Sciences) methods, to two manual methods, the QIAamp Viral RNA Mini kit (Qiagen) and TRIzol (Invitrogen), for the extraction of enterovirus RNA. Analytical sensitivity was assessed by dilution analysis of poliovirus type 2 Sabin in cerebrospinal fluid. The sensitivity of PCR was equivalent after RNA extraction with QIAamp, BioRobot M48, and MagNA Pure. All 18 replicates of 100 PFU/ml were detected after extraction by the four methods. Fewer replicates of each successive dilution were detected after extraction by each method. At 10(-1) PFU/ml, 17 of 18 replicates were positive by QIAamp, 15 of 18 replicates were positive by BioRobot M48, and 12 of 18 replicates were positive by MagNA Pure; at 10(-2) PFU/ml, 4 of 17 replicates were positive by QIAamp, 2 of 18 replicates were positive by BioRobot M48, and 0 of 18 replicates were positive by MagNA Pure. At 10(-3) PFU/ml, no replicates were detected. Evaluation of TRIzol was discontinued after nine replicates due to a trend of lower sensitivity (at 10(-3) PFU/ml eight of nine replicates were positive at 100 PFU/ml, four of nine replicates were positive at 10(-1) PFU/ml, and zero of nine replicates were positive at 10(-2) PFU/ml). Concordant results were obtained in 24 of 28 clinical specimens after extraction by all methods. No evidence of contamination was observed after extraction by automated instruments. The data indicate that the sensitivity of enterovirus PCR is largely similar after extraction by QIAamp, BioRobot M48, and MagNA Pure; a trend of decreased sensitivity was observed after TRIzol extraction. However, the results of enterovirus PCR were largely concordant in patient samples, indicating that the four extraction methods are suitable for detection of enteroviruses in clinical specimens. PMID- 12904352 TI - Comparison of enzyme-linked immunosorbent assay, immunoblotting, and PCR for diagnosis of toxoplasmic chorioretinitis. AB - Ocular toxoplasmosis is the major cause of posterior uvetis in European populations. The clinical diagnosis of toxoplasmic chorioretinitis is based upon ophthalmoscopic findings, which are often but not always typical. Laboratory testing is therefore important to confirm the etiology of the disease. In the present 2-year prospective study, the relative diagnostic sensitivities of the three analytical techniques (enzyme-linked immunosorbent assay [ELISA], immunoblotting, and PCR) were compared by using a group of patients (n = 19) with suspected ocular toxoplasmosis. The relative specificities of the three techniques were assessed by including two control groups of patients: one with nontoxoplasmic and noninflammatory ocular disease (n = 48) and the other with nontoxoplasmic and inflammatory ocular disease (n = 20). All 19 of the clinically suspect patients had serological evidence of exposure to Toxoplasma gondii: 17 had been previously infected, and 2 had current infection. The analysis of paired aqueous humor and serum samples by ELISA and immunoblotting revealed the local production of specific antibodies of the immunoglobulin G type in 63% (12 of 19) and 53% (10 of 19) of patients, respectively. PCR analysis of aqueous humor samples confirmed the presence of T. gondii DNA in 28% (5 of 18) of cases. When combined, ELISA, immunoblotting, and PCR findings confirmed the toxoplasmic origin of retinal lesions in 83% (15 of 18) of patients. The relative specificities of the three techniques were 89% for ELISA and immunoblotting and 100% for PCR. PMID- 12904353 TI - Outbreak of extended-spectrum beta-lactamase VEB-1-producing isolates of Acinetobacter baumannii in a French hospital. AB - Twelve clonally related and multidrug-resistant Acinetobacter baumannii isolates were recovered during a 4-month period from 12 patients hospitalized at the Valenciennes Hospital in France. Antibiograms determined by the double-disk diffusion technique on cloxacillin-containing plates detected a clavulanic acid inhibited extended-spectrum beta-lactamase (ESBL). PCR and sequencing identified the gene encoding the Ambler class A ESBL VEB-1. This gene was located on the chromosome and was part of a class 1 integron identical to that previously identified in Pseudomonas aeruginosa isolates from Thailand. Additionally, seven clonally related bla(VEB-1)-positive A. baumannii strains were identified in the immediate environment of the hospitalized patients. This is the first report of the ESBL VEB-1 in Acinetobacter spp. and the first description of VEB-1-producing strains as a source of an outbreak occurring outside Southeast Asia. This report underlines the difficulty of the identification of ESBLs in A. baumannii. PMID- 12904354 TI - Bacterial diversity in cases of lung infection in cystic fibrosis patients: 16S ribosomal DNA (rDNA) length heterogeneity PCR and 16S rDNA terminal restriction fragment length polymorphism profiling. AB - The leading cause of morbidity and mortality in cystic fibrosis (CF) patients stems from repeated bacterial respiratory infections. Many bacterial species have been cultured from CF specimens and so are associated with lung disease. Despite this, much remains to be determined. In the present study, we characterized without prior cultivation the total bacterial community present in specimens taken from adult CF patients, extracting DNA directly from 14 bronchoscopy or sputum samples. Bacterial 16S ribosomal DNA (rRNA) gene PCR products were amplified from extracted nucleic acids, with analyses by terminal restriction fragment length polymorphism (T-RFLP), length heterogeneity PCR (LH-PCR), and sequencing of individual cloned PCR products to characterize these communities. Using the same loading of PCR products, 12 distinct T-RFLP profiles were identified that had between 3 and 32 T-RFLP bands. Nine distinct LH-PCR profiles were identified containing between one and four bands. T-RFLP bands were detected in certain samples at positions that corresponded to pathogens cultured from CF samples, e.g., Burkholderia cepacia and Haemophilus influenzae. In every sample studied, one T-RFLP band was identified that corresponded to that produced by Pseudomonas aeruginosa. A total of 103 16S rRNA gene clones were examined from five patients. P. aeruginosa was the most commonly identified species (59% of clones). Stenotrophomonas species were also common, with eight other (typically anaerobic) bacterial species identified within the remaining 17 clones. In conclusion, T-RFLP analysis coupled with 16S rRNA gene sequencing is a powerful means of analyzing the composition and diversity of the bacterial community in specimens sampled from CF patients. PMID- 12904355 TI - Quality control trial for human immunodeficiency virus type 1 drug resistance testing using clinical samples reveals problems with detecting minority species and interpretation of test results. AB - Between January and March 2000, a quality control panel for human immunodeficiency virus (HIV) drug resistance testing was analyzed by 20 laboratories in five countries. The panel consisted of three clinical samples with different drug resistance genotypes and phenotypes and one HIV-negative plasma. Participants were asked to report the methods used for amplification and sequencing, a list of drug resistance-associated mutations that were detected in the protease and reverse transcriptase of each sample, and an interpretation concerning the susceptibility or resistance to 14 antiretroviral drugs. A total of 22 genotypic data sets were generated, which showed an overall good technical quality except for three participants, who failed to report key mutations for drug resistance. Problems were encountered in three respects: (i). resistant minorities of L90M in the protease, which were determined to about 12% by real time amplification, were only detected by one-fourth of the participants; (ii). newly described resistance mutations were frequently not reported; and (iii). interpretations of drug resistance-associated mutations varied widely, in particular for protease inhibitors. In some cases, different interpretations were caused by differences in the detection of resistant minorities, but even for the same genotypic profile, interpretations varied considerably. Similar discrepancies were revealed if current Web-based interpretation systems were used to predict drug resistance for samples of the proficiency panel. This indicates that a consensus for the interpretation of drug resistance-associated mutations is urgently needed. PMID- 12904356 TI - Evidence for genetic linkage between the gene segments encoding NSP4 and VP6 proteins in common and reassortant human rotavirus strains. AB - NSP4-encoding genes of 78 human rotavirus strains of common or reassortant genotypes were characterized by reverse transcription-PCR followed by sequencing and phylogenetic analysis. It was found that all the human strains characterized clustered into only two of the five known NSP4 genotypes. Linkage between NSP4 genotypes and VP6 subgroups was 100%, NSP4 genotype A being linked to VP6 of subgroup I (SGI) and NSP4 of genotype B being linked to VP6 of SGII. The diversity among the NSP4- and VP6-encoding genes was significantly less than that among the VP7 and VP4 genes in cocirculating human rotavirus strains. Whereas G and P types appear to be shared among different animal species and humans, the NSP4- and VP6-encoding genes appear to segregate according to their host of origin, suggesting that these two proteins may be host restriction determinants. The NSP4-VP6 association may be structurally determined during rotavirus replication (morphogenesis). PMID- 12904357 TI - Antimicrobial susceptibilities of salmonella strains isolated from humans, cattle, pigs, and chickens in the Netherlands from 1984 to 2001. AB - We monitored antimicrobial susceptibility data for Salmonella strains isolated from humans, cattle, pigs, and chickens in The Netherlands from 1984 to 2001 in order to provide insight into the dynamics of resistance over time. The strains were tested for their susceptibilities to seven antimicrobial agents by the agar diffusion method. Resistance was most common in Salmonella enterica subsp. enterica serovar Typhimurium. Among the strains from humans, pigs, and chickens, it was found that the level of resistance of serovar Typhimurium strains to tetracycline, ampicillin, chloramphenicol, and trimethoprim-sulfamethoxazole increased from 1984 to 2001. This increase could be attributed to the emergence of multidrug-resistant serovar Typhimurium DT 104. Among the strains from cattle, it was found that the level of resistance of serovar Typhimurium strains, which was already very high in the 1980s, declined during the study period to the same levels as those for the strains from the other species from 1996 to 2001. Serovar Enteritidis isolates remained susceptible during the entire survey period. Among serovar Paratyphi B variation Java strains isolated from chickens, resistance to furazolidone, flumequine, trimethoprim-sulfamethoxazole, and ampicillin emerged, although furazolidone was not used after 1990. Together, the data indicate that the levels and patterns of resistance differed considerably between Salmonella serovars isolated from one host species. PMID- 12904358 TI - Development and evaluation of a blocking enzyme-linked immunosorbent assay for detection of avian metapneumovirus type C-specific antibodies in multiple domestic avian species. AB - The first cases of infection caused by avian metapneumoviruses (aMPVs) were described in turkeys with respiratory disease in South Africa during 1978. The causative agent was isolated and identified as a pneumovirus in 1986. aMPVs have been detected in domestic nonpoultry species in Europe, but tests for the detection of these viruses are not available in the United States. To begin to understand the potential role of domestic ducks and geese and wild waterfowl in the epidemiology of aMPV, we have developed and evaluated a blocking enzyme linked immunosorbent assay (bELISA) for the detection of aMPV type C (aMPV-C) specific antibodies. This assay method overcomes the species-specific platform of indirect ELISAs to allow detection of aMPV-C-specific antibodies from potentially any avian species. The bELISA was initially tested with experimental turkey serum samples, and the results were found to correlate with those of virus neutralization assays and indirect enzyme-linked immunosorbent assay (iELISA). One thousand serum samples from turkey flocks in Minnesota were evaluated by our bELISA, and the level of agreement of the results of the bELISA and those of the iELISA was 94.9%. In addition, we were able to show that the bELISA could detect aMPV-C-specific antibodies from experimentally infected ducks, indicating its usefulness for the screening of serum samples from multiple avian species. This is the first diagnostic assay for the detection of aMPV-C-specific antibodies from multiple avian species in the United States. PMID- 12904359 TI - Genotyping and phenotyping of beta2-toxigenic Clostridium perfringens fecal isolates associated with gastrointestinal diseases in piglets. AB - Although Clostridium perfringens is recognized as an important cause of clostridial enteric diseases, only limited knowledge exists concerning the association of particular C. perfringens toxinotypes (type A to E) with gastrointestinal (GI) diseases in domestic animals. Some C. perfringens isolates also produce the newly discovered beta2-toxin (CPB2). Recent epidemiological studies suggested that C. perfringens isolates carrying the gene encoding CPB2 (cpb2) are strongly associated with clostridial GI diseases in domestic animals, including necrotic enteritis in piglets and typhlocolitis in horses. These putative relationships, obtained by PCR genotyping, were tested in the present study by further genotyping and phenotyping of 29 cpb2-positive C. perfringens isolates from pigs with GI disease (pig GI disease isolates). PCR and restriction fragment length polymorphism analysis reconfirmed the presence of cpb2 gene sequences in all the disease isolates included in the study. Furthermore, genotyping by pulsed-field gel electrophoresis analyses showed that the pig GI disease isolates included in this study all carry a plasmid cpb2 gene, yet no clonal relationships were detected between the cpb2-positive pig GI disease isolates surveyed. Finally, CPB2-specific Western blotting demonstrated CPB2 expression by all of the cpb2-positive isolates surveyed. The CPB2 proteins made by five of these pig GI disease isolates were shown to have the same deduced amino acid sequences as the biologically active CPB2 protein made by the original type C isolate, CWC245. Collectively, our present results support a significant association between CPB2-positive C. perfringens isolates and diarrhea in piglets. PMID- 12904360 TI - Detection of Chlamydia trachomatis and Neisseria gonorrhoeae by strand displacement amplification and relevance of the amplification control for use with vaginal swab specimens. AB - Vaginal swab specimens may be preferable to cervical swab or urine specimens for the detection of Chlamydia trachomatis and Neisseria gonorrhoeae because of the ease of specimen collection and transport. The purpose of this study was to evaluate whether vaginal swab specimens are equivalent to cervical swab specimens for the detection of N. gonorrhoeae and C. trachomatis by the Becton Dickinson strand displacement amplification assay (SDA) with the BDProbeTec ET instrument and then to evaluate the use of the amplification control in a clinical research setting. In the first phase, vaginal and cervical swab specimens were obtained from 455 symptomatic women aged 18 to 40 attending primary health care and sexually transmitted disease clinics. Thirty-nine specimens (8.6%) had true positive results for N. gonorrhoeae and 37 specimens (8.1%) had true-positive results for C. trachomatis. The sensitivity of SDA was superior to that of culture for the detection of N. gonorrhoeae with vaginal swab specimens and equivalent to that of the Roche PCR for the detection of C. trachomatis with cervical swab specimens. In the second phase of the study, 1,411 consecutively collected vaginal swab specimens were evaluated, with 357 (25.3%) specimens giving indeterminate readings on the basis of the result for the amplification control. The prevalences of sexually transmitted pathogens in vaginal swab specimens with and without use of the amplification control were 6.0 and 5.8%, respectively, for C. trachomatis and 3.1 and 3.0%, respectively, for N. gonorrhoeae. Although, vaginal swab specimens were equivalent to cervical swab specimens for the detection of N. gonorrhoeae and C. trachomatis by SDA with respect to sensitivity, one in four vaginal swab specimens yielded an indeterminate result when the amplification control was used. The amplification control has limited value for use with vaginal swab specimens. PMID- 12904361 TI - Comparison of SmartCycler real-time reverse transcription-PCR assay in a public health laboratory with direct immunofluorescence and cell culture assays in a medical center for detection of influenza A virus. AB - A single-tube real-time (fluorogenic) reverse transcription (RT)-PCR with the SmartCycler instrument (SmartCycler RT-PCR) for influenza A virus detection was evaluated with 238 respiratory specimens. Direct immunofluorescence antibody staining (DFA) and primary rhesus monkey kidney cell culture were performed on site at Yale-New Haven Hospital. Specimens were transported to the Connecticut Department of Public Health Laboratory for real-time RT-PCR. Cell culture detected influenza A virus in all 150 influenza A virus-positive specimens, DFA detected the virus in 148 influenza A virus-positive specimens, and SmartCycler RT-PCR detected the virus 143 influenza A virus-positive specimens. The sensitivity and specificity of RT-PCR were 95.3 and 100%, respectively. The high sensitivity and specificity and the rapid turnaround time made the SmartCycler RT PCR valuable for the rapid diagnosis of influenza A, especially in a public health laboratory. The closed real-time RT-PCR system avoided cross-contamination possible with RT-PCR and the excessive manipulations required for conventional RT PCR analysis and saved time and labor as well. In a medical center, rapid diagnosis by DFA was labor intensive but was 98.7% sensitive and 100% specific compared to the results of culture and provided results within 2 h throughout operating hours, helping with bed allocation on admission and patient management. PMID- 12904362 TI - Identification of swine hepatitis E virus (HEV) and prevalence of anti-HEV antibodies in swine and human populations in Korea. AB - The swine hepatitis E virus (HEV) is considered to be a new zoonotic agent due to its close genomic resemblance to the human HEV and its ability to infect nonhuman primates. Hepatitis caused by HEV infection has been a serious public health problem in developing countries. However, recent seroprevalence studies indicate that the HEV also circulates in industrialized countries. In this study, a nested reverse transcription (RT)-PCR was developed to detect a part of the swine HEV open reading frame 2. Three Korean isolates of swine HEV were identified in 128 swine sera (2.3% prevalence) by the nested RT-PCR method. They were isolated from 2- to 3-month old pigs showing an age-specific prevalence of the HEV viremia. A phylogenetic tree analysis with a number of swine and human HEV isolates indicated that all Korean isolates of the swine HEV belong to genotype III. They were closely related to the swine and human HEV isolates that were identified in the United States and Japan. In addition, they formed a distinct branch in genotype III, showing a 92.7 to 99.8% identity at their nucleotide sequences. The overall prevalence of anti-swine HEV antibodies in swine was 15%. Antibodies to the swine HEV were not detected in 1-month-old pigs. However, the anti-swine HEV antibodies appeared in pigs older than 1 month and also showed an age-specific prevalence. The antibody prevalence rates to the swine HEV were 6.0, 10.0, 36.0, and 25.0%, in 2-, 3-, 4-, and 5-to-7-month-old pigs, respectively. In addition, the seroprevalence in sows to the swine HEV was 8.8%. On the other hand, 18% of blood donors in Korea were found to be positive for anti-HEV antibodies. Overall, this study indicates that subclinical HEV infections may prevail in swine and human populations in Korea. PMID- 12904363 TI - Coagulase-negative staphylococci: comparison of phenotypic and genotypic oxacillin susceptibility tests and evaluation of the agar screening test by using different concentrations of oxacillin. AB - This study evaluated the oxacillin susceptibilities of 152 coagulase-negative staphylococcal (CoNS) strains of 12 species by disk diffusion; agar dilution; E test; the slide latex agglutination test (Slidex MRSA Detection test; bioMerieux S/A, Paris, France); the agar screening test with 1, 2, 4, or 6 microg of oxacillin per ml and incubation for 24 or 48 h; and detection of the mecA gene by PCR. The results revealed that the agar screening test with 4 micro g of oxacillin per ml and incubation for 48 h was superior to any single phenotype based susceptibility assay, presenting a sensitivity and a specificity of 100% each. For the different methods evaluated, the sensitivities and specificities were as follows: for disk diffusion, 94.2 and 91.8%, respectively; for the agar dilution test 100 and 73.5%, respectively; for E-test, 100 and 71.4%, respectively; and for the slide latex agglutination test, 97.1 and 98%, respectively. A good correlation was observed between oxacillin susceptibility testing results and PCR results for Staphylococcus epidermidis, S. haemolyticus, S. hominis subsp. hominis, and all mecA-positive strains. However, at least 60% of the mecA-negative isolates of the species S. saprophyticus, S. cohnii subsp. urealyticum, S. lugdunensis, and S. sciuri were erroneously classified as oxacillin resistant by the agar dilution test. Conversely, the slide latex agglutination test presented a high sensitivity (97.1%) and a high specificity (98%) for all CoNS species. Our results demonstrated the accuracy of the agar screening test with 4 micro g of oxacillin per ml and incubation for 48 h and the slide latex agglutination test for the appropriate detection of the oxacillin susceptibilities of CoNS isolates. Both assays are technically simple and can be easier to perform in routine laboratories than PCR. PMID- 12904365 TI - Activities of caspofungin, itraconazole, posaconazole, ravuconazole, voriconazole, and amphotericin B against 448 recent clinical isolates of filamentous fungi. AB - We examined the in vitro activity of caspofungin, posaconazole, voriconazole, ravuconazole, itraconazole, and amphotericin B against 448 recent clinical mold isolates. The endpoint for reading caspofungin was the minimum effective concentration (MEC). Among the triazoles, posaconazole was most active, inhibiting 95% of isolates at 95% of Aspergillus spp. at 20000, P = 0.003), presence of a central venous catheter (OR 2.0, P = 0.0002), and presence of anaerobic organism (OR 2.5, P = 0.04). Even after adjustments were made for underlying illness and length of stay, nosocomial status of bloodstream infection was strongly associated with increased total hospital charges (P < 0.0001). Although accounting for about half of all bloodstream infections, nosocomial bloodstream infections account for most of the mortality and costs associated with bloodstream infection. PMID- 12904370 TI - Genetic and antigenic characterization of rotavirus serotype G9 strains isolated in Australia between 1997 and 2001. AB - Rotavirus serotype G9 is recognized as the most widespread of the emerging serotypes, emerging since 1996 as a frequent cause of severe acute gastroenteritis in children from many countries covering all continents of the world. This study characterized serotype G9 strains collected in three widely separated Australian centers from 1997 to 2001. All G9 strains possessed the VP4 P[8] and VP6 subgroup II genes. The overall prevalence of the G9 strains increased in Australia, from 0.6% of the strains found in 1997 to 29% of the strains found in 2001. The prevalence of G9 relative to all other serotypes varied from year to year and with geographic location. In Melbourne (representing east coast urban centers), G9 made up 11 to 26% of all of the strains found from 1999 to 2001. In Perth (representing west coast urban centers), G9 made up less than 2% of the strains found in 1997 to 2000 but increased to 18.6% of the strains found in 2001. In Alice Springs (representing widely dispersed settlements in northern arid regions), G9 made up 0 to 5% of the strains found from 1997 to 2000 and was the dominant strain in 2001, making up 68.9% of all of the strains found. Three distinct antigenic groups based on reaction with neutralizing monoclonal antibodies (N-MAbs) were identified, including a dominant group (63%) that cross-reacted with the serotype G4 N-MAb. Phylogenetic analysis of the VP7-encoding gene from Australian strains, compared with a worldwide collection of G9 strains, showed that the Australian G9 strains made up a genetic group distinct from other serotype G9 strains identified in the United States and Africa. Future epidemiological studies of the occurrence of G9 strains should combine reverse transcription-PCR and typing with G1 to G4 and G9 N-MAbs to determine the extent of G9 and G4 cross-reactions among rotavirus strains, in order to assess the need to incorporate G9 strains into new candidate vaccines. PMID- 12904372 TI - Virus detection protocols for west nile virus in vertebrate and mosquito specimens. AB - The recent outbreaks of West Nile virus (WNV) infection in the northeastern United States and other regions of the world have made it essential to develop efficient, sensitive, and rapid protocols for virus surveillance. Laboratory testing is the backbone of any surveillance program. Protocols to detect the presence of WNV have been refined since 1999 for sensitivity, speed, efficiency, and specificity. This paper presents the protocols currently used by the New York State Department of Health to handle vertebrate and mosquito specimens that have been submitted for WNV testing to the Arbovirus Laboratories of the Wadsworth Center. PMID- 12904373 TI - Serodiagnosis of syphilis: antibodies to recombinant Tp0453, Tp92, and Gpd proteins are sensitive and specific indicators of infection by Treponema pallidum. AB - Syphilis serodiagnosis relies on a combination of nonspecific screening tests (antilipoidal antibodies) and Treponema pallidum-specific tests (anti-T. pallidum antibodies). We studied a group of six recombinant T. pallidum antigens for their sensitivities and specificities with sera from individuals with syphilis (n = 43), relapsing fever (n = 8), Lyme disease (n = 8), and leptospirosis (n = 9) and from uninfected individuals (n = 15). Three recombinant proteins, Tp0155, Tp0483, and Tp0751, demonstrated sensitivity values that ranged from 28 to 42%. In contrast, three other recombinant proteins exhibited the following sensitivity and specificity values: Tp0453, 100% sensitivity and 100% specificity; Tp92 (Tp0326), 98% sensitivity and 97% specificity; and Gpd (Tp0257), 91% sensitivity and 93% specificity. Tp0453, Tp92, and Gpd also were recognized by sera from individuals with early primary syphilis that were nonreactive with the antilipoidal Venereal Disease Research Laboratory test. The reactivities of syphilis patient sera with Tp0453, Tp92, and Gpd were proportional to the titers of these sera with the treponemal test MHA-TP (microhemagglutination assay for T. pallidum). Thus, the recombinant T. pallidum antigens Tp0453, Tp92, and Gpd show promise as diagnostic antigens in the enzyme-linked immunosorbent assay-based assay. PMID- 12904374 TI - Detection of circulating gp43 antigen in serum, cerebrospinal fluid, and bronchoalveolar lavage fluid of patients with paracoccidioidomycosis. AB - Paracoccidioidomycosis (PCM) is an important systemic fungal disease, particularly among individuals living and working in rural areas of endemicity in Latin America, who, without antifungal therapy, may develop fatal acute or chronic infection. For such patients, the detection of antibody responses by immunodiffusion is of limited value due to false-negative results. In contrast, the detection of Paracoccidioides brasiliensis gp43 circulating antigen may represent a more practical approach to the rapid diagnosis of the disease. Accordingly, an inhibition enzyme-linked immunosorbent assay (inh-ELISA) was developed for the detection of a 43-kDa P. brasiliensis-specific epitope incorporating a species-specific murine monoclonal antibody. With sera from patients with acute and chronic forms of the disease (n = 81), the overall sensitivity of the test was found to be 95.1%, while specificity was found to be 97.5% compared to that with normal human sera from blood donors (n = 93) and sera from patients with other chronic fungal infections (histoplasmosis [n = 33] and cryptococcosis [n = 20]). The inh-ELISA detected circulating antigen in 100% of patients with the acute form of PCM and in 95.31 and 100% of patients with the chronic multifocal and unifocal forms of PCM according to the patient's clinical presentation. Cerebrospinal fluid from 14 patients with neuroparacoccidioidomycosis and 13 samples of bronchoalveolar lavage fluid from patients with pulmonary unifocal PCM were also tested for gp43 detection, with the test showing 100% sensitivity and specificity. This novel, highly specific inh-ELISA represents a significant addition to the existing tests for the diagnosis of PCM. PMID- 12904375 TI - Secreted portion of glycoprotein g of herpes simplex virus type 2 is a novel antigen for type-discriminating serology. AB - The secreted portion of glycoprotein G (sgG-2) of herpes simplex virus type 2 (HSV-2) was evaluated as a novel antigen in an enzyme-linked immunosorbent assay (ELISA) format for detection of type-specific immunoglobulin G (IgG) antibodies in HSV-2-infected patients. The results were compared with those obtained by a commercially available assay, the HerpeSelect 2 ELISA (the FOCUS2 assay). Five different panels of sera were analyzed: panel A consisted of 109 serum samples from patients with a culture-proven HSV-1 infection that were Western blotting (WB) negative for HSV-2; panel B consisted of 106 serum samples from patients with a culture-proven recurrent HSV-2 infection that were WB positive for HSV-2; panel C consisted of 100 serum samples with no detectable IgG antibodies against HSV-1 and HSV-2; panel D consisted of 70 HSV-2 negative "tricky" serum samples containing antinuclear IgG antibodies or IgM antibodies against other viruses or bacteria; and panel E consisted of consecutive serum samples from 21 patients presenting with a first episode of HSV-2-induced lesions. When sera in panels A to C were analyzed, the sgG-2 ELISA and the FOCUS2 assay both showed sensitivities and specificities of >or=98%. In total, among the samples in panel D, 13 serum samples (19%) were false positive by the FOCUS2 assay and 1 serum sample (1.4%) was false positive by the sgG-2 ELISA. When the sera in panel E were analyzed, the sgG-2 ELISA detected seroconversion somewhat later than WB or the FOCUS2 assay did. We conclude that sgG-2 induces an HSV-2 type-specific antibody response and can be used for type-discriminating serology. PMID- 12904376 TI - New generation of cell culture assay for smallpox vaccine potency. AB - The potency of smallpox vaccines produced in the 1970s was tested by titration onto chorioallantoic membranes of fertilized hen eggs (CAM assay). The potency specification commonly approved for these vaccines was a titer above 10(8) pock forming units per milliliter. We developed and validated a cell culture titration assay to have a more reliable potency test. The cell titration assay and the CAM assay were tested in parallel on 34 first-generation smallpox vaccine lots. These allowed us to demonstrate that a correlation does exist between the two titration techniques and to determine a new in-house specification for the cell titration method. This in vitro potency assay will allow us to test first-generation smallpox vaccines produced on the skin of living animals but will also give a hint of the potency specification that should be assigned for new generations of cell-derived smallpox vaccines. PMID- 12904377 TI - PCR-reverse line blot typing method underscores the genomic heterogeneity of Borrelia valaisiana species and suggests its potential involvement in Lyme disease. AB - Detection of the Borrelia burgdorferi sensu lato complex in biological samples is currently done by conventional immunological and molecular biological methods. To improve on the accuracy of these methods and to simplify the procedure for testing large numbers of samples, a solid-phase sandwich hybridization system readily applicable to the detection of PCR products has been designed. This colorimetric detection system relies on the use of polybiotinylated detection probes and of specific capture oligonucleotides covalently linked at allocated positions on nylon membrane strips. From a phylogenetic analysis on a great number of ospA gene sequences, we have designed and synthesized a set of PCR primers specific to the five Borrelia burgdorferi sensu lato genospecies present in Europe and a subset of probes (capture and detection probes) specific to these five genospecies (B. burgdorferi sensu stricto, B. garinii, B. afzelii, B. valaisiana, and B. lusitaniae). This combined PCR hybridization system was evaluated with a large number of various B. burgdorferi isolates and clinical specimens. These analyses clearly showed that the system could be used as a typing method to distinguish five genospecies belonging to the B. burgdorferi sensu lato complex. In addition, the study showed that B. valaisiana strains might be more heterologous than suspected up to now and clustered into three genomic groups. PMID- 12904379 TI - Comparison of the Bordetella pertussis and Bordetella parapertussis isolates circulating in Saint Petersburg between 1998 and 2000 with Russian vaccine strains. AB - We analyzed the Bordetella pertussis and Bordetella parapertussis isolates circulating in Saint Petersburg that were collected between 1998 and 2000 and compared them with isolates collected 40 years ago and Russian vaccine strains. The analysis involved serotyping, pulsed-field gel electrophoresis of chromosomal DNA after digestion with XbaI and SpeI, and sequencing of the ptxS1 and prn genes, which encode the S1 subunit of the pertussis toxin and the major adhesin pertactin, respectively. The Russian isolates were classified in five of the six pulsed-field gel electrophoresis groups identified in other European countries. The B. pertussis isolates currently circulating in Saint Petersburg differed from the Russian whole-cell vaccine strains and the isolates collected in the prevaccine era. However, their repartition in the major pulsed-field gel electrophoresis groups was slightly different from that of isolates collected in countries that have had a high level of vaccine coverage for a long time, probably because the level of vaccine coverage in Saint Petersburg has increased only recently, after decreasing until the early 1990s. Most of the B. parapertussis isolates studied were similar to those circulating in France. However, some variants were observed, perhaps because B. parapertussis infections are more common in children in this area. PMID- 12904378 TI - Rapid and sensitive assays for determination of hepatitis B virus (HBV) genotypes and detection of HBV precore and core promoter variants. AB - Hepatitis B virus (HBV) genotypes may influence HBeAg seroconversion rates, mutational patterns in the precore (PC) and core promoter (CP) regions, severity of liver disease, and response to antiviral treatment. Development of rapid, simple, and standardized assays to detect viral genotypes and common mutations in the PC and CP regions can accelerate research on the clinical significance of these variants. We aim to assess the accuracy of a line probe assay in determining HBV genotypes and detecting HBV PC and CP variants. HBV genotypes in 701 patients and PC and CP variants in 600 patients with chronic HBV infection from China and the United States were studied using the INNO-LiPA assay. All but one (99.9%) sample were classified by the genotyping assay. All eight genotypes, i.e., A to H, were found. The INNO-LiPA genotyping assay results were completely concordant with those of sequencing. Using the INNO-LiPA PC assay, 99.8 and 94.7% samples were classifiable in the PC and CP regions, respectively. The PC assay results were completely concordant with those of sequencing in all samples that showed either wild-type or variant sequence. The line probe assay was more sensitive in detecting mixtures than was direct sequencing. By INNO-LiPA, only 50 and 27% of the samples, with mixed wild-type and variant sequence in the PC and CP region, respectively, showed mixed sequence by direct sequencing. INNO-LiPA is rapid, sensitive, and reliable-thus enabling accurate determination of HBV genotypes and detection of PC and CP variants in a large population of patients. PMID- 12904380 TI - Evaluation of the Osiris expert system for identification of beta-lactam phenotypes in isolates of Pseudomonas aeruginosa. AB - Osiris is a video zone size reader for disk diffusion tests featuring a built-in extended expert system (EES). The efficacy of the EES for the identification of the beta-lactam susceptibility phenotypes of Pseudomonas aeruginosa isolates was evaluated. Thirteen beta-lactams were tested in four laboratories by the disk diffusion test with 53 strains with well-characterized resistance mechanisms, including the production of 12 extended-spectrum beta-lactamases (ESBLs). The plates were read with the Osiris system and the results were interpreted with the ESS, and then the phenotype identified by the EES was compared to the resistance mechanism. The strains were also screened for the presence of ESBL production by a double-disk synergy test by placing the strains between an extended-spectrum cephalosporin-containing disk and a clavulanic acid-containing disk at distances of 30, 20, 15, and 10 mm from each other. Overall, the EES accurately identified the phenotypes of 88.2% of the strains and indicated an association with several mechanisms for 3.8% of the strains. No phenotype was identified in four strains with low levels of penicillinase production. Misidentifications were observed for two penicillinase-producing strains: one strain with partially derepressed cephalosporinase production and one strain overexpressing the MexA-MexB-OprM efflux system. The production of only four ESBLs was detected by the standard synergy test with a 30-mm distance between the disks. The production of five further ESBLs was identified by reducing the distance to 20 mm, and the production of the last three ESBLs was detected only at a distance of 15 or 10 mm. Our results indicate that the Osiris EES is an effective tool for the identification of P. aeruginosa beta-lactam phenotypes. A specific double-disk synergy test with reduced disk distances is necessary for the detection of ESBL production by this organism. PMID- 12904381 TI - Genomic approach to identification of Mycobacterium bovis diagnostic antigens in cattle. AB - Differential delayed-type hypersensitivity skin testing with tuberculin purified protein derivatives from Mycobacterium bovis and M. avium is the standard for diagnosing bovine tuberculosis. However, improved tests based on defined, specific antigens are urgently needed. In the present study, a combination of bioinformatics, molecular biology, and bovine models of infection were used to screen mycobacterial proteins for their potential as diagnostic reagents which could be used in a whole-blood assay for diagnosis of tuberculosis. Initial screening of 28 proteins selected in silico and expressed as recombinants in Escherichia coli indicated that CFP-10, ESAT-6, TB27.4, TB16.2, TB15.8, and TB10.4 induced strong gamma interferon responses in experimentally infected cattle. A more thorough investigation over time in two groups of animals infected with a high (10(6) CFU) and a low (10(4) CFU) dose of M. bovis revealed that, for both groups, the strength of the in vitro response to individual antigens varied greatly over time. However, combining the results for ESAT-6, CFP-10, and TB27.4, possibly supplemented with TB10.4, gave sensitivities at different infection stages close to those obtained with M. bovis purified protein derivative. Importantly, while responsiveness to ESAT-6 and CFP-10 correlated strongly for individual samples, the same was not the case for ESAT-6 and TB27.4 responsiveness. The results suggest that combinations of specific antigens such as these have great potential in development of optimized diagnostic systems for bovine tuberculosis. PMID- 12904382 TI - Oral Actinomyces isolates forming red colonies on brain heart blood agar can bee unambiguously classified as A odontolyticus by macroscopic examination. AB - The accurate classification of oral Actinomyces isolates as one species is difficult. Out of 18 Actinomyces isolates forming red colonies on brain heart blood agar, 12 could be straightforwardly assigned as Actinomyces odontolyticus by biochemical, morphological, and chemotaxonomic characteristics. For the remaining six isolates, the results of the different identification methods were inconsistent. By sequencing a 16S ribosomal DNA fragment by a rapid mass spectrometric method, all isolates could be identified unambiguously as A. odontolyticus. This result proves the importance of red colony pigmentation on brain heart blood agar together with the characteristic cell morphology for unequivocal assignment of oral Actinomyces isolates to the species A. odontolyticus. PMID- 12904383 TI - Genotypic identification of erythromycin-resistant campylobacter isolates as helicobacter species and analysis of resistance mechanism. AB - The correct identification of Campylobacter species remains cumbersome, especially when conventional biochemical tests and antimicrobial susceptibility patterns are used for a phenotypical identification. Correct identification is important for epidemiological purposes and for studying changes in antimicrobial resistance patterns. Six erythromycin-resistant campylobacter strains were investigated by 16S ribosomal DNA (rDNA) sequencing, 23S rDNA sequencing, and restriction fragment length polymorphism analysis of a putative heme-copper oxidase domain described as being specific for thermophilic Campylobacter species. Three erythromycin-resistant isolates from feces of human immunodeficiency virus (HIV)-seropositive patients with diarrhea and one blood isolate of from HIV-seropositive patient with cellulitis were identified by 16S rDNA analysis as Helicobacter cinaedi, whereas 23S rDNA sequencing suggested Wolinella succinogenes. The 16S rDNA sequence data of fecal isolates of two patients with travelers diarrhea revealed Helicobacter pullorum and were also in contrast with 23S rDNA sequencing. Of 4 H. cinaedi isolates, 1 contained the putative heme-copper oxidase gene thought to be specific for thermophilic species. The six erythromycin-resistant Helicobacter species had a similar point mutation A2143G in 23S rDNA resembling the macrolides resistance in Helicobacter pylori. We conclude that 16S rDNA sequencing should be preferred to 23S rDNA analysis and that macrolide-resistant campylobacter strains should be investigated by this approach for a correct identification. PMID- 12904384 TI - Virulence genes and neutral DNA markers of Helicobacter pylori isolates from different ethnic communities of West Bengal, India. AB - Virulence-associated genes and neutral DNA markers of Helicobacter pylori strains from the Santhal and Oroan ethnic minorities of West Bengal, India, were studied. These people have traditionally been quite separate from other Indians and differ culturally, genetically, and linguistically from mainstream Bengalis, whose H. pylori strains have been characterized previously. H. pylori was found in each of 49 study participants, although none had peptic ulcer disease, and was cultured from 31 of them. All strains carried the cag pathogenicity island and potentially toxigenic s1 alleles of vacuolating cytotoxin gene (vacA) and were resistant to at least 8 micro g of metronidazole per ml. DNA sequence motifs in vacA mid region m1 alleles, cagA, and an informative insertion or deletion motif next to cagA from these strains were similar to those of strains from ethnic Bengalis. Three mobile elements, IS605, IS607, and ISHp608, were present in 29, 19, and 10%, respectively, of Santhal and Oroan strains, which is similar to their prevalence in Bengali H. pylori. Thus, there is no evidence that the gene pools of H. pylori of these ethnic minorities differ from those of Bengalis from the same region. This relatedness of strains from persons of different ethnicities bears on our understanding of H. pylori transmission between communities and genome evolution. PMID- 12904385 TI - Enzootic transmission of Anaplasma bovis in Nantucket cottontail rabbits. AB - Serological studies of cottontail rabbits sampled from Nantucket Island, Mass., have suggested exposure to at least two ehrlichiae. The agent of human granulocytic ehrlichiosis (Anaplasma phagocytophilum) is intensely enzootic in rabbits there, but the identity of the other ehrlichial infection remains undescribed. We sampled rabbits over five transmission seasons and tested their blood and tissues for evidence of infection using PCR targeting an Ehrlichia genus-wide 16S rDNA target. Sequence analysis of positive amplicons revealed the presence of Anaplasma bovis, an agent not known to be present in North America. The average annual prevalence of A. bovis within rabbits, as determined by PCR of blood samples, was 18%. Haemaphysalis leporispalustris appears to serve as vector. The public health (human or veterinary) significance of this finding remains speculative. PMID- 12904386 TI - Geographic diversity among genotypes of Entamoeba histolytica field isolates. AB - It has been known that only 5 to 10% of those infected with Entamoeba histolytica develop symptomatic disease. However, the parasite and the host factors that determine the onset of disease remain undetermined. Molecular typing by using polymorphic genetic loci has been proven to aid in the close examination of the population structure of E. histolytica field isolates in nature. In the present study, we analyzed the genetic polymorphisms of two noncoding loci (locus 1-2 and locus 5-6) and two protein-coding loci (chitinase and serine-rich E. histolytica protein [SREHP]) among 79 isolates obtained from different geographic regions, mainly Japan, Thailand, and Bangladesh. When the genotypes of the four loci were combined for all isolates that we have analyzed so far (overlapping isolates from mass infection events were excluded), a total of 53 different genotypes were observed among 63 isolates. The most remarkable and extensive variations among the four loci was found in the SREHP locus; i.e., 34 different genotypes were observed among 52 isolates. These results demonstrate that E. histolytica has an extremely complex genetic structure independent of geographic location. Our results also show that, despite the proposed transmission of other sexually transmitted diseases, including human immunodeficiency virus infection, from Thailand to Japan, the spectra of the genotypes of the E. histolytica isolates from these two countries are distinct, suggesting that the major E. histolytica strains prevalent in Japan at present were likely introduced from countries other than Thailand. Although the genetic polymorphism of the SREHP locus was previously suggested to be closely associated with the clinical presentation, e.g., colitis or dysentery and liver abscess, no association between the clinical presentation and the SREHP genotype at either the nucleotide or the predicted amino acid level was demonstrated. PMID- 12904387 TI - Monitoring of cytomegalovirus infection in solid-organ transplant recipients by an ultrasensitive plasma PCR assay. AB - Early and accurate monitoring of cytomegalovirus (CMV) infection in solid-organ transplant recipients is of major importance. We have assessed the potential benefit of an ultrasensitive plasma-based PCR assay for renal transplant recipients. The pp65 CMV antigen (pp65 Ag) assay using leukocytes was employed as a routine test for the monitoring of CMV in 23 transplant recipients. We compared the pp65 antigenemia with the CMV load quantified by an ultrasensitive PCR (US PCR) with a limit of detection of 20 CMV DNA copies/ml of plasma. CMV infection was detected in 215 (67%) of 321 plasma samples by the US-PCR compared with 124 (39%) of 321 samples by the pp65 Ag assay. The US-PCR assay permitted the detection of CMV infection episodes following transplantation a median of 12 days earlier than the pp65 Ag assay. Moreover, during CMV infection episodes, DNA detection by the US-PCR was consistently positive, whereas false negative results were frequently observed with the pp65 Ag assay. We found a good correlation between the two assays, and the peak viral loads were significantly higher in patients with CMV-related complications (median, 5000 DNA copies/ml) than in those without symptoms (1160 DNA copies/ml) (P = 0.048). In addition, patients that did not require preemptive therapy based on the results of the pp65 assay had CMV loads significantly lower (median, 36 DNA copies/ml) than those that needed treatment (median, 4703 DNA copies/ml) (P < 0.001). These observations provided cutoff levels that could be applied in clinical practice. The ultrasensitive plasma-based PCR detected CMV infection episodes earlier and provided more consistent results than the pp65 Ag assay. This test could improve the monitoring of CMV infection or reactivation in renal transplant recipients. PMID- 12904388 TI - Optimization and validation of multilocus sequence typing for Candida albicans. AB - Multilocus sequence typing (MLST) was applied to 75 Candida albicans isolates, including 2 that were expected to be identical, 48 that came from diverse geographical and clinical sources, and 15 that were sequential isolates from two patients. DNA fragments ( approximately 500 bp) of eight genes encoding housekeeping functions were sequenced, including four that have been described before for C. albicans MLST, and four new gene fragments, AAT1a, AAT1b, MPI, and ZWF1. In total, 87 polymorphic sites were found among 50 notionally different isolates, giving 46 unique sequence types, underlining the power of MLST to differentiate isolates for epidemiological studies. Additional typing information was obtained by detecting variations in size at the transcribed spacer region of the 25S rRNA gene and tests for homozygosity at the mating type-like (MTL) locus. The stability of MLST was confirmed in two sets of consecutive isolates from two patients. In each set the isolates were identical or varied by a single nucleotide. Reference strain SC5314 and a derived mutant, CAF2, gave identical MLST types. Heterozygous polymorphisms were found in at least one isolate for all but 16 (18.4%) of the variable nucleotides, and 35 (41%) of the 87 individual sequence changes generated nonsynonymous amino acids. Cloning and restriction digestion of a gene fragment containing heterozygous polymorphisms indicated that the heterozygosity was genuine and not the result of sequencing errors. Our data validate and extend previous MLST results for C. albicans, and we propose an optimized system based on sequencing eight gene fragments for routine MLST with this species. PMID- 12904389 TI - Presence of activatable Shiga toxin genotype (stx(2d)) in Shiga toxigenic Escherichia coli from livestock sources. AB - Stx2d is a recently described Shiga toxin whose cytotoxicity is activated 10- to 1000-fold by the elastase present in mouse or human intestinal mucus. We examined Shiga toxigenic Escherichia coli (STEC) strains isolated from food and livestock sources for the presence of activatable stx(2d). The stx(2) operons of STEC were first analyzed by PCR-restriction fragment length polymorphism (RFLP) analysis and categorized as stx(2), stx(2c vha), stx(2c vhb), or stx(2d EH250). Subsequently, the stx(2c vha) and stx(2c vhb) operons were screened for the absence of a PstI site in the stx(2A) subunit gene, a restriction site polymorphism which is a predictive indicator for the stx(2d) (activatable) genotype. Twelve STEC isolates carrying putative stx(2d) operons were identified, and nucleotide sequencing was used to confirm the identification of these operons as stx(2d). The complete nucleotide sequences of seven representative stx(2d) operons were determined. Shiga toxin expression in stx(2d) isolates was confirmed by immunoblotting. stx(2d) isolates were induced for the production of bacteriophages carrying stx. Two isolates were able to produce bacteriophages phi1662a and phi1720a carrying the stx(2d) operons. RFLP analysis of bacteriophage genomic DNA revealed that phi1662a and phi1720a were highly related to each other; however, the DNA sequences of these two stx(2d) operons were distinct. The STEC strains carrying these operons were isolated from retail ground beef. Surveillance for STEC strains expressing activatable Stx2d Shiga toxin among clinical cases may indicate the significance of this toxin subtype to human health. PMID- 12904390 TI - Vaginal swabs are appropriate specimens for diagnosis of genital tract infection with Chlamydia trachomatis. AB - Because self-collected vaginal swabs (VS) are potentially very useful for screening asymptomatic women for Chlamydia trachomatis infection, a multicenter study evaluated that specimen with nucleic acid amplification tests (NAATs). The objective was to determine whether VS are equal to Food and Drug Administration (FDA)-cleared specimens (cervical swabs and first-catch urines [FCU]) for diagnosing genital chlamydial infection. All NAATs then commercially available (October 1996 to October 1999) were used (ligase chain reaction [LCx Probe System; Abbott Laboratories, Abbott Park, Ill.]; PCR [Amplicor; Roche Molecular Systems, Branchburg, N.J.]; and transcription-mediated amplification, [Amplified CT Assay; Gen-Probe Inc., San Diego, Calif.]). NAATs were performed on FCU, urethral, cervical, self- and clinician-collected VS. Sensitivity was compared to isolation using cervical and urethral swabs. Agreement of NAAT results between VS and cervical swabs or FCU was calculated. Specimens from 2,517 15- to 25-year-old asymptomatic women attending clinics at nine different centers were evaluated. Results with self- and clinician-collected VS were equivalent and were at least as good as results with FCU and cervical swabs. Across all sites, summary specificities for all specimens were >99%. Among culture-positive women, NAAT sensitivity with VS (93%) was as high as or higher than NAAT sensitivity with cervical swabs (91%) or FCU (80.6%) or culture of cervical swabs (83.5%). VS are appropriate specimens for diagnosing chlamydial genital tract infection by NAATs. That patients can efficiently collect them offers important benefits for screening programs. It would be beneficial for public health programs if the NAAT manufacturers sought FDA clearance for this specimen. PMID- 12904391 TI - Rapid identification of bacteria from positive blood cultures by terminal restriction fragment length polymorphism profile analysis of the 16S rRNA gene. AB - Bacteremia results in significant morbidity and mortality, especially among patient populations that are immunocompromised. Broad-spectrum antibiotics are administered to patients suspected to have bloodstream infections that are awaiting diagnosis that depends on blood culture analysis. Significant delays in identification of pathogens can result, primarily due to the dependence on growth based identification systems. To address these limitations, we took advantage of terminal restriction fragment (TRF) length polymorphisms (T-RFLP) due to 16S ribosomal DNA (rDNA) sequence diversity to rapidly identify bacterial pathogens directly from positive blood culture. TRF profiles for each organism were determined by sizing fragments from restriction digests of PCR products derived from two sets of 16S rDNA-specific fluorescent dye-labeled primers. In addition, we created a TRF profile database (TRFPD) with 5899 predicted TRF profiles from sequence information representing 2860 different bacterial species. TRF profiles were experimentally determined for 69 reference organisms and 32 clinical isolates and then compared against the predicted profiles in the TRFPD. The predictive value of the profiles was found to be accurate to the species level with most organisms tested. In addition, identification of 10 different genera was possible with profiles comprising two or three TRFs. Although it was possible to identify Enterobacteriaceae by using a profile of three TRFs, the similarity of the TRF profiles of these organisms makes differentiation of species less reliable with the current method. The ability to rapidly (i.e., within approximately 8 h) identify bacteria from blood cultures has potential for reducing unnecessary use of broad-spectrum antibiotics and promoting more timely prescription of appropriate antibiotics. PMID- 12904392 TI - Evidence of parvovirus replication in cerebral neurons of cats. AB - The correlation between parvovirus infections and lesions in the central nervous system other than cerebellar hypoplasia was studied in 100 cats. The animals were necropsied with a history of various diseases, one third showing typical clinical and pathomorphological signs of panleukopenia. In 18 cats polyclonal antiserum against canine parvovirus consistently labeled neurons mainly in diencephalic regions, whereas the cerebellar cortex remained negative in all cases. In situ hybridization with digoxigenin-labeled minus-sense RNA probes, hybridizing with monomer-replicative form DNA or mRNA, revealed positive signals in nuclei of several neurons of the brain, again excluding the cerebellum. PCR applied to formalin-fixed and paraffin-embedded brain tissue and intestinal tissues of the diseased cats and subsequent DNA sequence analysis yielded canine parvovirus type 2 (CPV-2)-like sequences in the central nervous system. Two aspects of these findings are intriguing: (i). parvoviruses appear to be capable of replicating in neurons, cells that are considered to be terminally differentiated and (ii). CPV like viruses of the old antigenic type CPV-2 appear to be able to infect cats. PMID- 12904394 TI - Culture of Tropheryma whipplei from human samples: a 3-year experience (1999 to 2002). AB - The culture of Tropheryma whipplei, the bacterium responsible for Whipple's disease, has been established only recently. Our objective is to describe, based on our experience, the culture of T. whipplei in HEL cells detected by immunofluorescence staining. Over 3 years, we received 18 samples for T. whipplei culture from 15 patients with Whipple's disease. Ten duodenal biopsy specimens from 10 patients with digestive symptoms were available. Five cardiac valves and three blood samples from five patients with endocarditis were also available. We correlated the results of culture with the type of sample and the culture procedure. Seven isolates were obtained, and three were subsequently established for more than 4 passages. The mean delay for the primary detection was 30 days. The bacterium was isolated more frequently from sterile specimens (5 of 8) than from duodenal biopsy specimens (2 of 10), but the difference (P = 0.14) was not significant. Decontamination of digestive samples containing colistin, amphotericin B, and cephalotin or ciprofloxacin did not impair the isolation of T. whipplei. The use of vancomycin precludes the primary isolation (7 of 12 versus 0 of 6; P = 0.08) and the establishment of T. whipplei (3 of 12 versus 0 of 6; P = 0.5). Omitting samples cultured with vancomycin, the establishment of the strain was significantly higher when antibiotics were prescribed for no more than 7 days (3 of 4 versus 0 of 8; P = 0.03). Our results demonstrate that samples must be collected within 1 week of an antibiotic regimen's initiation for the successful establishment of the bacterium. PMID- 12904393 TI - Evolution of sporadic isolates of methicillin-resistant Staphylococcus aureus (MRSA) in hospitals and their similarities to isolates of community-acquired MRSA. AB - Forty-one methicillin-resistant Staphylococcus aureus (MRSA) hospital isolates that clearly differed from the six major pandemic clones of MRSA in pulsed-field gel electrophoresis type, mecA and Tn554 polymorphism, and epidemic behavior were selected from an international strain collection for more detailed characterization. SpaA typing, multilocus sequence typing, and SCCmec (staphylococcal cassette chromosome mec) typing demonstrated extensive diversity among these sporadic isolates both in genetic background and also in the structure of the associated SCCmec elements. Nevertheless, the isolates could be grouped into restricted clonal complexes by using the BURST (i.e., based upon related sequence types) program algorithm, which predicted that most sporadic MRSA isolates evolved from pandemic MRSA clones. Several of the sporadic MRSA resembled community-acquired MRSA isolates in properties that included a relatively limited multiresistance pattern, faster growth rates, diversity of genetic backgrounds, and a frequent association with SCCmec type IV. PMID- 12904395 TI - Differences in gene content among Salmonella enterica serovar typhi isolates. AB - We used a nonredundant microarray of the Salmonella enterica serovar Typhimurium LT2 and Typhi CT18 genomes to assess the genomic content of a diverse set of isolates of serovar Typhi. Comparative genomic hybridization revealed 13 regions of absent or divergent gene content in the eight Typhi strains examined compared to Typhi CT18. In particular, two Typhi CT18 prophage regions, STY1048 to STY1077 and STY2038 to STY2077, as well as a five-gene islet (STY3188 to STY3193) were absent or divergent in all other Typhi strains examined. Seven Typhi strains lacked most or all of the IS1 elements present in strain CT18, and three Typhi strains lacked a P4-like phage (STY4821 to STY4834). One strain was devoid of a 149-gene region (STY4521 to STY4680), which encodes numerous phage genes and the Vi antigen biosynthesis and export gene cluster, a type IV pilus, and numerous phage genes. In Typhi strain 26T25, an amplification of an entire inter-ribosomal region encompassing 31 genes has occurred. Furthermore, a 257-gene region (STY1360 to STY1639) showed an aberrant replication pattern in three Typhi isolates. Overall, these differences in gene content indicate that even within a highly clonal bacterial population the genomic reservoir is unstable. PMID- 12904397 TI - Real-time PCR assay to detect smallpox virus. AB - We developed a highly sensitive and specific assay for the rapid detection of smallpox virus DNA on both the Smart Cycler and LightCycler platforms. The assay is based on TaqMan chemistry with the orthopoxvirus hemagglutinin gene used as the target sequence. With genomic DNA purified from variola virus Bangladesh 1975, the limit of detection was estimated to be approximately 25 copies on both machines. The assay was evaluated in a blinded study with 322 coded samples that included genomic DNA from 48 different isolates of variola virus; 25 different strains and isolates of camelpox, cowpox, ectromelia, gerbilpox, herpes, monkeypox, myxoma, rabbitpox, raccoonpox, skunkpox, vaccinia, and varicella zoster viruses; and two rickettsial species at concentrations mostly ranging from 100 fg/ microl to 1 ng/ microl. Contained within those 322 samples were variola virus DNA, obtained from purified viral preparations, at concentrations of 1 fg/ microl to 1 ng/ microl. On the Smart Cycler platform, 2 samples with false positive results were detected among the 116 samples not containing variola virus tested; i.e., the overall specificity of the assay was 98.3%. On the LightCycler platform, five samples with false-positive results were detected (overall specificity, 95.7%). Of the 206 samples that contained variola virus DNA ranging in concentrations from 100 fg/ microl to 1 ng/ microl, 8 samples were considered negative on the Smart Cycler platform and 1 sample was considered negative on the LightCycler platform. Thus, the clinical sensitivities were 96.1% for the Smart Cycler instrument and 99.5% for the LightCycler instrument. The vast majority of these samples were derived from virus-infected cell cultures and variola virus infected tissues; thus, the DNA material contained both viral DNA and cellular DNA. Of the 43 samples that contained purified variola virus DNA ranging in concentration from 1 fg/ microl to 1 ng/ microl, the assay correctly detected the virus in all 43 samples on both the Smart Cycler and the LightCycler platforms. The assay may be useful for the early detection of smallpox virus infections should such infections occur as a result of a deliberate or an accidental recurrence. PMID- 12904396 TI - Transmission of Anaplasma marginale by Boophilus microplus: retention of vector competence in the absence of vector-pathogen interaction. AB - Whether arthropod vectors retain competence for transmission of infectious agents in the long-term absence of vector-pathogen interaction is unknown. We addressed this question by quantifying the vector competence of two tick vectors, with mutually exclusive tropical- versus temperate-region distributions, for genetically distinct tropical- and temperate-region strains of the cattle pathogen Anaplasma marginale. The tropical cattle tick Boophilus microplus, which has been eradicated from the continental United States for over 60 years, was able to acquire and transmit the temperate St. Maries (Idaho) strain of A. marginale. Similarly, the temperate-region tick Dermacentor andersoni efficiently acquired and transmitted the Puerto Rico strain of A. marginale. There were no significant quantitative differences in infection rate or number of organisms per tick following feeding on cattle with persistent infections of either A. marginale strain. In contrast, the significantly enhanced replication of the Puerto Rico strain in the salivary gland of B. microplus at the time of transmission feeding is consistent with adaptation of a pathogen strain to its available vector. However, the transmission of both strains by B. microplus demonstrates that adaptation or continual interaction between the pathogen and vector is not required for retention of vector competence. Importantly, the results clearly show that reestablishment of acaricide-resistant B. microplus in the United States would be associated with A. marginale transmission. PMID- 12904398 TI - Automated extraction and quantification of human cytomegalovirus DNA in whole blood by real-time PCR assay. AB - The measurement of human cytomegalovirus (HCMV) DNA in blood is becoming the standard method for monitoring HCMV infection in immune-suppressed and unsuppressed patients. As various blood compartments can be used, we have compared the HCMV DNA measured in whole blood (WB), peripheral blood leukocytes (PBL), and plasma by real-time PCR. We tested 286 samples: HCMV DNA was extracted automatically from WB and PBL with the MagNA Pure instrument (Roche Molecular Biochemicals) and manually from plasma samples. The HCMV DNA from WB, PBL, and plasma was measured by real-time Light Cycler PCR. Primers and probe were located in the UL 83 region. HCMV DNA was detected more frequently in WB (88.5%) than in the PBL (65.7%) (P < 0.0001) or the plasma (55.2%) (P < 0.0001). There was a good correlation between the positive results in WB and in PBL (r = 0.68; P < 0.0001), and 3.15 log(10) genome copies in 200000 PBL, equivalent to the threshold value of 50 pp65-positive polymorphonuclear cells per 200000 leukocytes, was equivalent to 3.4 log(10) genome copies in 200 microl of WB. WB was shown to be suitable for automated extraction and the quantitation of HCMV DNA by real-time Light Cycler PCR by analysis of serial samples from representative patients of various populations. This system may be very useful for monitoring of immune-suppressed and unsuppressed patients. PMID- 12904399 TI - Evaluation of a modified single-enzyme amplified-fragment length polymorphism technique for fingerprinting and differentiating of Mycobacterium kansasii type I isolates. AB - The usefulness of single-enzyme amplified-fragment length polymorphism (AFLP) analysis for the subtyping of Mycobacterium kansasii type I isolates was evaluated. This simplified technique classified 253 type I strains into 12 distinct clusters. The discriminating power of this technique was high, and the technique easily distinguished between the epidemiologically unrelated control strains and our clinical isolates. Overall, the technique was relatively rapid and technically simple, yet it gave reproducible and discriminatory results. This technique provides a powerful typing tool which may be helpful in solving many questions concerning the reservoirs, pathogenicities, and modes of transmission of these isolates. PMID- 12904400 TI - Evaluation of a rapid PCR assay for diagnosis of meningococcal meningitis. AB - We compared the results of Gram staining and culture of cerebrospinal fluid to results obtained with a rapid PCR assay for the diagnosis of meningococcal meningitis in 281 cases of suspected bacterial meningitis. PCR had a sensitivity of 97% compared to a sensitivity of 55% for culture, and the PCR specificity was 99.6%. PCR results were available within 2 h of the start of the assay. PMID- 12904401 TI - Type III secretion genes in Aeromonas salmonicida subsp salmonicida are located on a large thermolabile virulence plasmid. AB - Type III secretion genes in Aeromonas salmonicida subsp. salmonicida are located on a large plasmid of approximately 140 kb. Cultivation of this organism at elevated temperatures such as 25 degrees C can, however, result in loss of this plasmid. This is accompanied by a loss of virulence for cultured fish cells. PMID- 12904402 TI - Genetic diversity among Escherichia coli O157:H7 isolates from Bovines living on farms in England and Wales. AB - Pulsed-field gel electrophoresis of Escherichia coli O157:H7 isolates (n = 228) from 122 healthy animals on 11 farms discriminated 57 types. Most clones were found only on individual farms. Numerous clones were found within each farm, with a prevalent clone normally found in several animals. A variety of clones were found within the different phage types. PMID- 12904403 TI - Rapid identification of Candida glabrata with a new commercial test, GLABRATA RTT. AB - The GLABRATA RTT test (Fumouze Diagnostics, Levallois Perret, France) is based on the ability of Candida glabrata to hydrolyze trehalose but not maltose. It requires an inoculum of only four to six colonies, and the results are available within 20 min. We tested GLABRATA RTT with 330 stock isolates grown in subcultures on four different primary fungal isolation media and obtained a sensitivity of 94 to 98% (depending on the medium used) and a specificity of 97.3 to 98.6%. The false-positive results corresponded to C. tropicalis, C. famata, and C. lusitaniae. GLABRATA RTT thus offers rapid and reliable identification of C. glabrata. PMID- 12904404 TI - Improved molecular detection of dengue virus serotype 1 variants. AB - The dengue virus molecular typing method described by Lanciotti and coworkers (R. S. Lanciotti, C. H. Calisher, D. J. Gubler, G. J. Chang, and A. Vance-Vorndam, J. Clin. Microbiol. 30:545-551, 1992) is used worldwide for diagnosis and surveillance. However, it failed to detect DENV-1 variants in Cambodia due to a point mutation. We describe an improvement of the method that allows the detection of additional DENV-1 strains, including potential variants. PMID- 12904405 TI - Evaluation of the rapid immunoassay determine HIV 1/2 for detection of antibodies to human immunodeficiency virus types 1 and 2. AB - We evaluated the reliability of a rapid human immunodeficiency virus type 1 test for quick clinical decision making, such as in needle-stick accidents. The test was evaluated with 1,160 patients. It proved to be a simple and useful test with 99.6% specificity and 99.4% sensitivity. One patient with late-stage AIDS had a false-negative result. PMID- 12904406 TI - Quantitative real-time PCR assay for detection of Ehrlichia chaffeensis. AB - A real-time PCR assay was developed for the detection of Ehrlichia chaffeensis. The assay is species specific and provides quantitative results in the range 10 to 10(10) gene copies. The assay is not inhibited by the presence of tick, human, or mouse DNA and is compatible with high sample throughput. The assay was compared with previously described assays for E. chaffeensis. PMID- 12904407 TI - Identification of the hypervirulent lineages of Neisseria meningitidis, the ST-8 and ST-11 complexes, by using monoclonal antibodies specific to NmeDI. AB - Most cases of serogroup C meningococcal disease are caused by the clonal lineages ST-8 and ST-11. The gene encoding the putative restriction-modification system NmeDI is specific to these lineages. We report here a monoclonal antibody directed against the NmeDI endonuclease as a tool for their rapid and spe-cific identification. PMID- 12904408 TI - Phylogenetic analysis of hemoplasma species: an international study. AB - Nearly complete 16S rRNA gene sequences for feline and canine hemoplasma isolates from Europe, Australia, Africa, and Asia showed almost 100% identity to those previously reported for United States isolates. Partial sequences of the RNA subunit of the RNase P gene were also determined, and RNase P-based phylogenetic analysis showed that the hemoplasmas are most closely related to the members of the Mycoplasma pneumoniae group. PMID- 12904409 TI - Monitoring response to antiviral therapy for patients with chronic hepatitis C virus infection by a core-antigen assay. AB - A recently released immunoassay detecting total serum hepatitis C virus (HCV) core antigen was used to prospectively monitor virological responses to antiviral treatment in patients with chronic HCV infection. Sustained responders cleared core protein from serum within the first month of therapy and maintained stably negative values for the entire duration of follow-up after treatment discontinuation. However, patients who relapsed or failed to respond showed transient negative values and could not be accurately discriminated either because of the intrinsic lower sensitivity of the core-antigen assay than those of molecular assays or because of differentially regulated secretion of immunoreactive core protein from infected hepatocytes. PMID- 12904410 TI - Identification of Exophiala mesophila isolated from treated dental unit waterlines. AB - Members of the genus Exophiala are often difficult to identify to the species level because of their variable morphological appearances. This paper describes the methods used to identify Exophiala mesophila and provides salient differential features for distinguishing other mesophilic members of the genus. PMID- 12904411 TI - Detection of methicillin-resistant Staphylococcus aureus (MRSA) in blood with the EVIGENE MRSA detection kit. AB - A total of 200 blood cultures containing putative staphylococci were analyzed by a commercial gene probe hybridization assay (EVIGENE; Statens Serum Institut, Copenhagen, Denmark), and 18 were identified as methicillin-resistant Staphylococcus aureus (MRSA) positive. Of these, 17 were positive by PCR and 16 were positive by culture. Detailed analysis of the discrepant results showed that the EVIGENE kit allowed specific identification of MRSA in blood cultures without any of the drawbacks associated with PCR. PMID- 12904412 TI - VIM-1 Metallo-beta-lactamase-producing Klebsiella pneumoniae strains in Greek hospitals. AB - Seventeen Klebsiella pneumoniae clinical isolates carrying the bla(VIM-1) metallo beta-lactamase gene were collected in the intensive care units of three hospitals in Athens, Greece, in 2002. They exhibited various carbapenem resistance levels (Etest MICs of imipenem ranged from 4 to 32 microg/ml). All isolates gave positive results by the imipenem-EDTA synergy Etest. The isolates were classified into four main types by pulsed-field gel electrophoresis; the majority of the isolates (5 and 10 isolates) belonged to two types. The bla(VIM-1) gene cassette was part of the variable region of a class 1 integron that also included aac6, dhfrI, and aadA. This structure was carried by transferable plasmids. PMID- 12904413 TI - Evaluation of susceptibility of human herpesvirus 8 to antiviral drugs by quantitative real-time PCR. AB - A new in vitro system based on real-time PCR was developed for evaluation of human herpesvirus 8 susceptibility to antiviral agents. Cidofovir had the greatest inhibitory activity against HHV-8 (50% inhibitory concentration [IC(50)], 0.43 microM) followed by ganciclovir (2.61 microM), adefovir (18.00 microM), acyclovir (31.00 microM), and foscarnet (34.15 microM). The potential therapeutic efficacy for HHV-8 (i.e., peak serum drug level/IC(50)) is highest for cidofovir (167) and foscarnet (22). PMID- 12904414 TI - Molecular cloning, expression, self-assembly, antigenicity, and seroepidemiology of a genogroup II norovirus isolated in France. AB - Virus-like particles of Dijon171/96 virus, a genogroup II norovirus, were expressed in a baculovirus system and were used for a seroepidemiological study of 1078 age-stratified human sera collected in Dijon, France. The results showed a seroprevalence of 74.1%. Furthermore, we showed that murine antibodies generated against recombinant Dijon171/96 virus, and human antibodies recognized discontinuous epitopes on the particles. PMID- 12904415 TI - Analysis of the ability of spirochete species associated with relapsing fever, avian borreliosis, and epizootic bovine abortion to bind factor H and cleave c3b. AB - Some Borrelia species associated with Lyme disease bind the complement-regulatory protein factor H (fH), a process that may aid in immune evasion. In this report we demonstrate that some Borrelia species associated with relapsing fever bind fH, but not those associated with avian borreliosis and epizootic bovine abortion. Cell-bound fH was also found to mediate cleavage of exogenously supplied human C3b, demonstrating the biological relevance of fH binding and its possible importance in the pathogenesis of the relapsing-fever spirochetes. PMID- 12904416 TI - Reproducibility of positive test results in the BDProbeTec ET system for detection of Chlamydia trachomatis and Neisseria gonorrhoeae. AB - Nucleic acid amplification tests such as the BDProbeTec ET (BDPT) system are more prone to reproducibility problems than are antigen detection tests and culture. A repeat testing algorithm for all samples with method other than acceleration (MOTA) scores greater than or equal to the cutoff value (2000) was developed for the BDPT system and applied in a clinical laboratory setting. All positive samples were retested, and if the result of the second test was below the cutoff value, a third test was performed to resolve the discrepancy. Overall, 11 (5.3%) of 207 samples initially positive for Chlamydia trachomatis and 11 (10.7%) of 103 samples initially positive for Neisseria gonorrhoeae were not confirmed by repeat testing of the original sample. Poor reproducibility was associated with low positive MOTA scores (2000 to 9999) for both analytes. Only 21 (80.8%) of 26 low positive samples in the C. trachomatis test and 4 (33.3%) of 12 low-positive samples in the N. gonorrhoeae test retested as positive. The reproducibility of both tests with samples with initial MOTA scores of >or=10000 increased to 96.7%. The data suggest that retesting of low-positive samples is warranted and could reduce the number of potentially false-positive test results. PMID- 12904417 TI - Molecular epidemiology of Haemophilus influenzae type b isolated from children with clinical cases of conjugate vaccine failures. AB - We analyzed eight invasive Haemophilus influenzae strains isolated from individual children vaccinated appropriately for their ages. Five of the strains were type b and three were nontypeable strains. Children infected with nontypeable strains had a protective level of anti-H. influenzae type b antibodies, while only one of the children whose cases represented true vaccine failure did. PMID- 12904418 TI - Dengue disease spectrum among infants in the 2001 dengue epidemic in Chennai, Tamil Nadu, India. AB - An investigation in a referral pediatric hospital has indicated that during a recent dengue outbreak in Chennai, Tamil Nadu, India, dengue in infancy constituted 20% of total dengue virus infections with low mortality rates in this hospital. In developing countries, strengthening of dengue management capabilities at hospitals can prevent dengue-related deaths in infants. PMID- 12904419 TI - Comparison of real-time PCR, conventional PCR, and galactomannan antigen detection by enzyme-linked immunosorbent assay using bronchoalveolar lavage fluid samples from hematology patients for diagnosis of invasive pulmonary aspergillosis. AB - An iCycler iQ real-time PCR assay targeting 18S rRNA Aspergillus-specific sequences was developed for the diagnosis of invasive pulmonary aspergillosis (IPA). Positive findings were obtained for 18 of 20 (90%) bronchoalveolar lavage (BAL) fluid specimens from patients with probable or confirmed IPA and were obtained for none of the 24 BAL samples from patients with no clinical evidence of aspergillosis. These results were concordant with those of a nested PCR assay, which detected 90% of the patients with IPA, while galactomannan ELISA revealed positivity for 100% of these patients, suggesting that combined use of methods might improve the diagnosis of IPA. PMID- 12904420 TI - Molecular typing of Campylobacter jejuni isolates involved in a neonatal outbreak indicates nosocomial transmission. AB - Genotypic typing by restriction fragment length polymorphism and pulsed-field gel electrophoresis showed that two neonates in a neonatal ward were infected with the same Campylobacter jejuni strain. Isolates from the mother and brother of the index patient were identical to each other but distinct from the neonatal type. Genotyping results therefore suggested that the neonatal C. jejuni infection was nosocomial in origin. PMID- 12904421 TI - Genomic analysis of Mycobacterium tuberculosis complex strains used for production of purified protein derivative. AB - The genomes of the tuberculin production strains Mycobacterium bovis AN5 and Mycobacterium tuberculosis DT were compared to genome-sequenced tubercle bacilli by using DNA microarrays. Neither the AN5 nor DT strain suffered extensive gene deletions during in vitro passage. This suggests that bovine tuberculin made from M. bovis AN5 is suitable to detect infection with presently prevalent M. bovis strains. PMID- 12904422 TI - Modification of the double-disk test for detection of enterobacteriaceae producing extended-spectrum and AmpC beta-lactamases. AB - Detection of extended-spectrum beta-lactamases (ESBLs) in AmpC-producing Enterobacteriaceae is problematic. A modification of the double-disk test (MDDT) has been developed for successful detection of ESBLs in gram-negative bacilli producing well-characterized beta-lactamases as well as 212 clinical isolates of Enterobacter cloacae, Enterobacter aerogenes, Serratia marcescens, and Citrobacter freundii. MDDT accurately differentiated between ESBL producers and derepressed chromosomal AmpC mutants. MDDT provides a cost-effective alternative approach for clinical microbiology laboratories for routine susceptibility testing with simultaneous detection of ESBLs in enterobacteriaceae. PMID- 12904423 TI - Strains of Streptococcus pyogenes from severe invasive infections bind HEp2 and HaCaT cells more avidly than strains from uncomplicated infections. AB - Epidemiologically unrelated Streptococcus pyogenes strains isolated from blood, throat, and skin were assayed for adherence to HEp2 and HaCaT cells. Invasive isolates showed significantly higher avidity for these cell lines than isolates from skin and throat. In general, S. pyogenes showed greater binding to HaCaT cells than to HEp2 cells. PMID- 12904424 TI - Use of dipsticks for rapid diagnosis of cholera caused by Vibrio cholerae O1 and O139 from rectal swabs. AB - We evaluated the recently developed dipsticks for the rapid detection of Vibrio cholerae serotypes O1 and O139 from rectal swabs of hospitalized diarrheal patients after enrichment for 4 h in alkaline peptone water. The sensitivity and specificity of the dipsticks were above 92 and 91%, respectively. The dipsticks represent the first rapid test which has been successfully used to diagnose cholera from rectal swabs, and this would immensely improve surveillance for cholera, especially in remote settings. PMID- 12904425 TI - Clinical impact of a PCR assay for identification of Staphylococcus aureus and determination of methicillin resistance directly from blood cultures. AB - We evaluated the clinical usefulness of a PCR assay that discriminates Staphylococcus aureus from coagulase-negative staphylococci and detects methicillin resistance on blood cultures by measuring the adaptation of antimicrobial therapy based on the PCR results. Only 7 of 28 patients (25%) benefited from a modification of antibiotic therapy based on the PCR results, since empirical therapy was appropriate in a majority of cases. PMID- 12904426 TI - Close relationship between G8-serotype bovine and human rotaviruses isolated in Nigeria. AB - A bovine rotavirus, NGRBg8, isolated from the feces of a calf with diarrhea in Nigeria was characterized by reverse transcription-PCR, nucleotide sequence analysis, and Northern blot hybridization. The nucleotide sequence of the VP7 gene of the strain was most closely related to that of a Nigerian human G8 serotype strain, HMG035 (99.9%). The NSP1 gene of strain NGRBg8 is highly related (99.4%) to that of a Thai G8 bovine strain, A5-10. Northern blot hybridization revealed a high overall genomic relatedness of bovine strain NGRBg8 with human strain HMG035; all 11 RNA segments hybridized to each other. Thus, the results show the close relationship between G8 bovine and human rotaviruses in Nigeria. PMID- 12904427 TI - Ability of lysozyme and 2-deoxyglucose to differentiate human and bovine Streptococcus bovis strains. AB - Human and bovine Streptococcus bovis strains had the same 16S ribosomal DNA restriction fragment length polymorphism and often had the same patterns of starch, mannitol, lactose, and raffinose utilization. PCRs of BOX sequences differed, but numerical analyses indicated that some human strains clustered with bovine strains. However, human and bovine strains had distinctly different sensitivities to lysozyme and 2-deoxyglucose. PMID- 12904428 TI - Vertical transmission of the hepatitis C virus to infants of anti-human immunodeficiency virus-negative mothers: molecular evolution of hypervariable region 1 in prenatal and perinatal or postnatal infections. AB - In a prospective study of 33 infants born to hepatitis C virus (HCV)-positive human immunodeficiency virus-negative mothers the vertical transmission of HCV occurred in 6.8%. The evolution of HCV infection in two babies was studied from birth up to 5 or 6 years of age, and the sequencing of the hypervariable region (HVR) of the putative envelope-encoding E2 region of the HCV genome was performed. The HVR1 sequence variability and the different serological profiles during follow-up could reflect the differences in HCV transmission routes, HCV genotypes, and clinical evolution of infection. PMID- 12904429 TI - Detection of the emerging rotavirus G9 serotype at high frequency in Italy. AB - Group A human rotavirus strains belonging to the unusual serotype G9 were detected at high frequency in stool specimens from infected children with acute diarrhea in Bari, Italy, during a 15-month survey from March 2001 to June 2002. This may signify a local reemergence of the G9 rotaviruses detected in Italy in the early and mid-1990s or may be related to the global emergence of G9 rotaviruses in recent years. PMID- 12904430 TI - babA2- and cagA-positive Helicobacter pylori strains are associated with duodenal ulcer and gastric carcinoma in Brazil. AB - The babA2 and cagA genes were investigated in 208 Brazilian Helicobacter pylori strains. A strong association between babA2 and duodenal ulcer or gastric carcinoma was observed, even after adjusting for confounding factors, such as age, gender, and cagA status. cagA-positive strains were also independently associated with H. pylori-related diseases. PMID- 12904431 TI - Commercial broth microdilution panel validation and reproducibility trials for garenoxacin (BMS-284756), a novel desfluoroquinolone. AB - Results from garenoxacin dry-form broth microdilution MIC panels prepared commercially (Sensititre, TREK Diagnostics) were compared to reference frozen form MICs to ensure the validity of the longer-shelf-life product. A total of 1078 organisms from seven major organism groups were used in this trial. All commercial MIC results were within +/- one log(2) dilution of reference garenoxacin values, and reproducibility trials produced identical MIC results for 90.5 to 92.1% of garenoxacin MIC comparisons. Control quinolones (ciprofloxacin and gatifloxacin) also performed at a similarly high level of accuracy. PMID- 12904432 TI - Genetic relationship between Streptococcus pneumoniae isolates from nasopharyngeal and cerebrospinal fluid of two infants with Pneumococcal Meningitis. AB - The molecular epidemiology of Streptococcus pneumoniae isolates from carriage and cerebrospinal fluid (CSF) concurrently recovered from the same individual has not yet been reported. By using pulsed-field gel electrophoresis, we demonstrated the genetic linkage among strains from CSF and nasopharynges of two children with pneumococcal meningitis. PMID- 12904433 TI - Seronegative bacteremic melioidosis caused by Burkholderia pseudomallei with ambiguous biochemical profile: clinical importance of accurate identification by 16S rRNA gene and groEL gene sequencing. AB - An aerobic gram-negative bacterium was isolated from the blood and sputum of an 84-year-old, chair-bound nursing home resident with acute bacteremic pneumonia. Although the phenotypic characteristics suggested that the bacterium could be Burkholderia pseudomallei, the Vitek 1 system (GNI+), which can successfully identify 99% of B. pseudomallei strains, showed that the bacterium was "unidentified." Immunoglobulin G against the lipopolysaccharide (LPS) of B. pseudomallei, as detected by an LPS-based enzyme-linked immunosorbent assay with 95% sensitivity, was negative in both the acute-phase and convalescent-phase sera. Sequencing of the groEL gene showed that the isolate was B. pseudomallei. Proper identification of the bacterium in this study is crucial, since there would be a radical difference in the duration of antimicrobial therapy. PMID- 12904434 TI - Acute cytomegalovirus infection and transient carotid intimal-medial thickening in a young, otherwise healthy woman. AB - Carotid intimal-medial thickening was observed in a 23-year-old woman with acute cytomegalovirus (CMV) infection. The thickening disappeared after her recovery from the infection. As endothelial cells are common targets of CMV, this thickening suggests that CMV infection causes vascular lesions, even in otherwise healthy individuals. PMID- 12904436 TI - Severe Serratia liquefaciens sepsis following vitamin C infusion treatment by a naturopathic practitioner. AB - A 66-year-old female patient developed severe Serratia liquefaciens sepsis following vitamin C infusion treatment by a naturopathic practitioner. The clinical course of the infection was characterized by several complications, and the direct costs of the hospital stay amounted to about 40000 Euro. Genotypically identical S. liquefaciens was isolated from the residue of the infusate given to the patient, as well as from the washbasin overflow and from two other infusion bottles. A careful inspection of the dispensing facilities and review of procedures used to prepare the infusate revealed several indications of poor hygiene. However, the source of contamination could not be fully clarified. This case report raises questions about the local facilities and personal qualifications required for naturopathic practitioners to conduct invasive procedures and demonstrates that lapses in hygiene can lead to severe morbidity and high cost. PMID- 12904437 TI - Novel nosocomial infections by Stenotrophomonas maltophilia: first reported case from Lucknow, North India. AB - We describe a case of empyema with infected ascites caused by Stenotrophomonas maltophilia, which has rarely been reported as pathogenic. The source was determined to a disinfectant solution. The isolate was sensitive to a newer carbapenem-meropenem, and the patient was treated successfully. This case represents a novel dual presentation of a nosocomial infection by the isolate in question. PMID- 12904435 TI - Scedosporium prolificans osteomyelitis in an immunocompetent child treated with voriconazole and caspofungin, as well as locally applied polyhexamethylene biguanide. AB - Scedosporium species are increasingly isolated from immunocompromised and immunocompetent patients. Unfortunately, Scedosporium infections are generally resistant to amphotericin B, and Scedosporium prolificans strains are particularly resistant to the antifungal agents now in use. We report here on an immunocompetent child with S. prolificans-associated osteomyelitis successfully treated with debridement, local irrigation with polyhexamethylene biguanide, and the systemic administration of voriconazole and caspofungin despite poor in vitro activity of voriconazole alone against the isolate. We also review the treatments and outcomes of 28 reported cases of osteomyelitis or septic arthritis caused by Scedosporium species in immunocompetent patients. PMID- 12904438 TI - Cat scratch disease presenting as orbital abscess and osteomyelitis. AB - Ocular manifestations of cat scratch disease are uncommon. The diagnosis is usually made on the basis of increasing Bartonella henselae serum antibody titers. We report a child presenting with orbital abscess and osteomyelitis who was diagnosed with hepatosplenic cat scratch disease by detection of B. henselae DNA in the orbital abscess fluid. PMID- 12904439 TI - Hymenolepis diminuta infection in a child living in the urban area of Rome, Italy. AB - We report a case of Hymenolepis diminuta infection in an Italian child affected by tuberous sclerosis. Praziquantel is the drug of choice for the treatment of H. diminuta infection. However, considering the patient's neurological disease, we decided to use not praziquantel but niclosamide, which proved equally effective. PMID- 12904441 TI - Rhizobium (Agrobacterium) radiobacter identified as a cause of chronic endophthalmitis subsequent to cataract extraction. AB - Herein, we report a case of chronic endophthalmitis caused by a ceftazidime resistant Rhizobium radiobacter strain in a 62-year-old male. The patient underwent an uneventful cataract extraction of the right eye a week prior to the appearance of symptoms (pain, redness, and blurring vision) which developed following a golf outing. Upon admission the patient received an emergency vitrectomy. The patient remained symptomatic, and R. radiobacter was isolated repeatedly from vitreous fluid cultures over a 5-month period. Ultimately, the infection responded to intravitreal gentamicin, oral ciprofloxacin, and removal of the lens implant. PMID- 12904440 TI - Fatal acute cellulitis due to Neisseria meningitidis. AB - We describe the first fatal evolution of cellulitis due to Neisseria meningitidis serogroup Y involving an 85-year-old woman. She presented with an extensive cellulitis of the left side of the face, neck, and thorax and septic shock. In spite of active antibiotic therapy, evolution was rapidly fatal. PMID- 12904442 TI - First isolation and detection by immunofluorescence assay of Bartonella koehlerae in erythrocytes from a French cat. PMID- 12904443 TI - Failure to detect Salmonella enterica serovar Dublin on Aes Laboratoire Salmonella Agar Plate. PMID- 12904444 TI - Distribution of fsr among Enterococcus faecalis isolates from the SENTRY antimicrobial surveillance program. PMID- 12904445 TI - Importance of primer specificity for PCR detection of Anaplasma phagocytophila among Ixodes scapularis ticks from Wisconsin. PMID- 12904446 TI - Granada agar sensitivity and detection of group B streptococcus. PMID- 12904447 TI - Blood agar, chocolate agar, and Mycobacterium tuberculosis. PMID- 12904448 TI - Proteomic changes during disturbance of cholesterol metabolism by azacoprostane treatment in Caenorhabditis elegans. AB - Although nematodes like Caenorhabditis elegans are incapable of de novo cholesterol biosynthesis, they can utilize nonfunctional sterols by converting them into cholesterol and other sterols for cellular function. The results reported previously and presented here suggest that blocking of sterol conversion to cholesterol in C. elegans by 25-azacoprostane-HCl (azacoprostane) treatment causes a serious defect in germ cell development, growth, cuticle development, and motility behavior. To establish a biochemical basis for these physiological abnormalities, we performed proteomic analysis of mixed stage worms that had been treated with the drug. Our results from a differential display proteomic analysis revealed significant decreases in the levels of proteins involved in collagen and cytoskeleton organization such as protein disulfide isomerase (6.7-fold), beta tubulin (5.41-fold), and NEX-1 protein (>30-fold). Also reduced were enzymes involved in energy production such as phosphoglycerate kinase (4.8-fold) and phosphoenolpyruvate carboxykinase (8.5-fold), a target for antifilarial drugs such as azacoprostane. In particular, reductions in the expression of lipoprotein families such as vitellogenin-2 (7.7-fold) and vitellogenin-6 (5.4-fold) were prominent in the drug-treated worms, indicating that sterol metabolism disturbance caused by azacoprostane treatment is tightly coupled with suppression of the lipid transfer-related proteins at the protein level. However, competitive quantitative reverse transcriptase polymerase chain reaction showed that the transcriptional levels of vit-2, vit-6, and their receptors (e.g. rme-2 and lrp 1) in drug-treated worms were 3- to 5-fold higher than those in the untreated group, suggesting a presence of a sterol regulatory element-binding protein (SREBP)-like pathway in these genes. In fact, multiple predicted sterol regulatory elements or related regulatory sequences responding to sterols were found to be located at the 5'-flanking regions in vit-2 and lrp-1 genes, and their transcriptional activities fluctuated highly in response to changes in sterol concentration. Thus, many physiological abnormalities caused by azacoprostane-mediated sterol metabolism disturbance appear to be exerted at least in part through SREBP pathway in C. elegans. PMID- 12904449 TI - Assessment of behavioral mechanisms maintaining encopresis: Virginia Encopresis Constipation Apperception Test. AB - OBJECTIVE: To develop and test a scale for parent and child, evaluating theoretical and clinical parameters relevant to children with encopresis. Encopretic children were hypothesized to have more bowel-specific, but not more generic, psychological problems, as compared with nonsymptomatic control children. In addition, mothers were also believed to be more discerning than children. METHODS: The Virginia Encopresis-Constipation Apperception Test (VECAT) consists of 9 pairs of bowel-specific and 9 parallel generic drawings. Respondents selected the picture in each pair that best described them/their child. It was administered to encopretic children (N = 87), nonsymptomatic siblings (N = 27), and nonsymptomatic nonsiblings (N = 35). The mothers of all the participants also completed the VECAT. Encopretic children were retested 6 and 12 months posttreatment with Enhanced Toilet Training. RESULTS: The VECAT demonstrated good test-retest reliability and internal consistency. Encopretic children and their mothers reported more bowel-specific, but not more generic, problems. Bowel-specific scores improved significantly posttreatment only for those patients who demonstrated significant symptom improvement. Mothers were significantly more discerning than children. CONCLUSION: The VECAT is a reliable, valid, discriminating, and sensitive test. Bowel-specific problems appear to best differentiate children with and without encopresis. PMID- 12904450 TI - The Illness Management Survey: identifying adolescents' perceptions of barriers to adherence. AB - OBJECTIVE: To develop a self-report measure of barriers to adherence and to evaluate its reliability and validity in a sample of adolescents with asthma. METHODS: The Illness Management Survey (IMS) was developed through item generation, expert panel review, and focus group administration. Adolescents with asthma (N = 152) completed the measure. Participants reported on perceived drawbacks to medication, risk-taking behavior, and social desirability tendencies. Providers rated adolescents' illness severity and adherence. Reliability and validity of the IMS were assessed, and factor structure was examined. RESULTS: The 27-item IMS shows high internal consistency (alpha =.87). Scores correlate with perceived medication drawbacks, risk taking, and self- and provider reports of adherence. Principal-components analysis indicates five domains of barriers, accounting for 52.4% of the variance: disease/regimen issues, cognitive difficulties, lack of social support/lack of self-efficacy, denial/distrust, and peer/family issues. CONCLUSIONS: Preliminary data indicate that the IMS reliably and validly assesses perceived barriers to adherence within this sample of adolescents with asthma. It shows promise as a tool for identifying subgroups of nonadherent adolescents. PMID- 12904451 TI - Congruence between parents' and adolescents' reports of special health care needs in a Title XXI program. AB - OBJECTIVES: The purpose of this study was to examine agreement between adolescents and their parents about whether or not the adolescent had a special health care need, using the Children with Special Health Care Needs (CSHCN) Screener. METHODS: Telephone surveys that included the CSHCN Screener were conducted with 522 adolescents and their parents who were new enrollees in Florida's State Children's Health Insurance Program (SCHIP). RESULTS: Analyses revealed substantial agreement as to whether or not the adolescent had a chronic condition. However, a full 15% of pairs disagreed. Analyses of pair disagreement revealed that parents reported adolescents' chronic conditions more often than adolescents, most strikingly for mental health conditions. Additional analyses revealed that pairs with older adolescents, female adolescents, and Hispanic origin had higher odds of being congruent than their counterparts. CONCLUSIONS: The results showed higher congruence using the consequence-based CSHCN Screener than is typically reported for diagnosis-based approaches. Despite an impressive rate of agreement, the analyses also highlighted parents' tendency to overreport special health care needs relative to their adolescent, particularly for mental health issues, and illustrated some of the demographic factors that might predict congruence. These findings are relevant to work related to the use of tools such as the CSHCN Screener in profiling enrollees in health care programs that serve children and adolescents. PMID- 12904452 TI - Stable preterm infants gain more weight and sleep less after five days of massage therapy. AB - OBJECTIVE: To examine the effects of 5 days of massage therapy on the weight gain and sleep/wake behavior of hospitalized stable preterm infants. METHODS: Massage therapy (body stroking/passive limb movement for three 15-minute periods per day) was provided to 16 preterm neonates (mean gestational age, 30.1 weeks; mean birth weight, 1359 g), and their weight gain, formula intake, kilocalories, stooling, and sleep/wake behavior were compared with a group of 16 control infants (mean gestational age, 31.1 weeks; mean birth weight, 1421 g). RESULTS: The massage group averaged 53% greater daily weight gain than the control group. The massage group spent less time sleeping at the end of 5 treatment days than the control group and more time in the drowsy state. CONCLUSIONS: Healthy, low-risk preterm infants gained more weight and slept less with just 5 days of massage, in contrast to 10 days in previous studies. Results support the continued use of massage as a cost-effective therapy for medically stable preterm infants. PMID- 12904453 TI - The Children's Somatization Inventory: further evidence for its reliability and validity in a pediatric and a community sample of Dutch children and adolescents. AB - OBJECTIVE: To examine the psychometric properties of the Children's Somatization Inventory (CSI) in the Netherlands. METHOD: The CSI and a number of personality and psychopathology questionnaires were administered to Dutch schoolchildren (N = 479), children referred to a pediatric clinic (N = 63), and children's parents. RESULTS: Factor analysis yielded a number of factors that have also been found in previous research, viz., pain/weakness, gastrointestinal symptoms, and pseudoneurological symptoms. The reliability (internal consistency) of the CSI was satisfactory. Furthermore, support was obtained for the validity of the CSI. More specifically, the scale correlated in a theoretically meaningful way with child and parent reports of personality and psychopathology, and discriminated well between healthy and pediatric children. Finally, highly similar psychometric properties were obtained for the Parent version of the CSI (i.e., PCSI). CONCLUSION: The Dutch version of the CSI seems to be a reliable and valid self report measure for assessing somatization symptoms in children and adolescents. PMID- 12904454 TI - Inpatient pediatric consultation-liaison: a case-controlled study. AB - OBJECTIVE: To conduct a prospective case-controlled study of pediatric inpatients referred for consultation in a tertiary care children's medical center. METHOD: Referrals (n = 104) were matched with nonreferrals (n = 104) for age (4 to 18 years), gender, and illness type/severity and completed parent- and self-report (dependent on age) behavioral rating scales to assess for adjustment/functioning. Nurses completed in-hospital ratings of behavioral/adjustment difficulties. Goal attainment and satisfaction ratings were obtained from the referring physicians, parents/guardians, and the consultant. RESULTS: Referrals exhibited more behavior/adjustment/coping difficulties than nonreferrals by parent, nurse, and self report. Frequently employed interventions included coping-strategies intervention, cognitive and behavioral therapies, and case management. Referring physician and consultant ratings of goal attainment were high, as were physician ratings of satisfaction and parent/guardian ratings of overall helpfulness. CONCLUSIONS: Pediatric inpatients referred by their physicians had significantly more internalizing and externalizing disturbances than their nonreferred hospitalized peers. Many of the behavioral and adjustment problems that lead to in-hospital consultation referral were evident in global behavior difficulties prior to hospitalization. Referring pediatricians, parents/guardians, and consultants rate the outcome as benefiting the patients via assisting in the overall management of their health concerns, coping, and adjustment. PMID- 12904455 TI - Training in pediatric psychology: a survey of predoctoral internship programs. AB - OBJECTIVE: To describe internship training in pediatric psychology and to determine its correspondence with the recommendations of the Society of Pediatric Psychology (SPP) Task Force on Training. METHODS: A survey based on the Task Force recommendations was sent to all internship programs that reported offering a major rotation in pediatrics. RESULTS: Opportunities in a wide range of assessment and intervention strategies were available. Training in disease process, medical management, and consultation/liaison with a variety of disciplines was also offered. Interns interacted with individuals from a range of developmental stages and ethnicities. Many professional issues were addressed. Experiences in research methods and prevention were less well represented. CONCLUSIONS: Despite some variability, experiences in a wide range of areas were offered by many programs in several of the training domains recommended by the SPP Task Force. However, there is room for improvement in training in research and prevention and health promotion. PMID- 12904456 TI - Commentary: training in pediatric psychology: a survey of predoctoral internship programs. PMID- 12904457 TI - Prolonged photoresponses and defective adaptation in rods of Gbeta5-/- mice. AB - Timely deactivation of G-protein signaling is essential for the proper function of many cells, particularly neurons. Termination of the light response of retinal rods requires GTP hydrolysis by the G-protein transducin, which is catalyzed by a protein complex that includes regulator of G-protein signaling RGS9-1 and the G protein beta subunit Gbeta5-L. Disruption of the Gbeta5 gene in mice (Gbeta5-/-) abolishes the expression of Gbeta5-L in the retina and also greatly reduces the expression level of RGS9-1. We examined transduction in dark- and light-adapted rods from wild-type and Gbeta5-/- mice. Responses of Gbeta5-/- rods were indistinguishable in all respects from those of RGS9-/- rods. Loss of Gbeta5-L (and RGS9-1) had no effect on the activation of the G-protein cascade, but profoundly slowed its deactivation and interfered with the speeding of incremental dim flashes during light adaptation. Both RGS9-/- and Gbeta5-/- responses were consistent with another factor weakly regulating GTP hydrolysis by transducin in a manner proportional to the inward current. Our results indicate that a complex containing RGS9-1-Gbeta5-L is essential for normal G-protein deactivation and rod function. In addition, our light adaptation studies support the notion than an additional weak GTPase-accelerating factor in rods is regulated by intracellular calcium and/or cGMP. PMID- 12904458 TI - Reversible paired helical filament-like phosphorylation of tau is an adaptive process associated with neuronal plasticity in hibernating animals. AB - Neurofibrillary pathology [paired helical filaments (PHFs)] formed by the microtubule-associated protein tau in a hyperphosphorylated form is a major hallmark of Alzheimer's disease and related disorders. The process of tau phosphorylation, thought to be of critical importance for PHF formation, and its potential link to neurodegeneration, however, is not understood very well, mostly because of the lack of a physiological in vivo model of PHF-like tau phosphorylation. Here we describe the formation of highly phosphorylated tau, containing a number of PHF-like epitopes in torpor during hibernation. PHF-like phosphorylation of tau was not associated with fibril formation and was fully reversible after arousal. Distribution of PHF-like tau followed a consistent pattern, being most intense in the entorhinal cortex, hippocampus, and isocortical areas. Within the hippocampus, a particularly high labeling was seen in CA3 pyramidal cells. Somewhat lesser reactivity was present in CA1 neurons while dentate gyrus granule cells were not reactive. Formation of PHF-like tau in CA3 neurons was paralleled by the regression of synaptic contacts of the mossy fiber system terminating on CA3 apical dendrites. Mossy fiber afferentation was re-established during arousal, concomitantly with the decrease of PHF-like tau in CA3 neurons. These findings implicate an essential link between neuronal plasticity and PHF-like phosphorylation of tau. The repeated formation and degradation of PHF-like tau might, thus, represent a physiological mechanism not necessarily associated with pathological effects. Hibernation will, therefore, be a valuable model to study the regulation of PHF-like tau-phosphorylation and its cell biological sequelae under physiological in vivo conditions. PMID- 12904459 TI - Multisensory integration during motor planning. AB - When planning goal-directed reaches, subjects must estimate the position of the arm by integrating visual and proprioceptive signals from the sensory periphery. These integrated position estimates are required at two stages of motor planning: first to determine the desired movement vector, and second to transform the movement vector into a joint-based motor command. We quantified the contributions of each sensory modality to the position estimate formed at each planning stage. Subjects made reaches in a virtual reality environment in which vision and proprioception were dissociated by shifting the location of visual feedback. The relative weighting of vision and proprioception at each stage was then determined using computational models of feedforward motor control. We found that the position estimate used for movement vector planning relies mostly on visual input, whereas the estimate used to compute the joint-based motor command relies more on proprioceptive signals. This suggests that when estimating the position of the arm, the brain selects different combinations of sensory input based on the computation in which the resulting estimate will be used. PMID- 12904460 TI - Methylprednisolone increases neuronal apoptosis during autoimmune CNS inflammation by inhibition of an endogenous neuroprotective pathway. AB - Optic neuritis is one of the most common clinical manifestations of multiple sclerosis (MS), a chronic inflammatory disease of the CNS. High-dosage methylprednisolone treatment has been established as the standard therapy of acute inflammation of the optic nerve (ON). The rationale for corticosteroid treatment lies in the antiinflammatory and immunosuppressive properties of these drugs, as shown in experimental autoimmune encephalomyelitis (EAE), the animal model of MS. To investigate the influence of methylprednisolone therapy on the survival of retinal ganglion cells (RGCs), the neurons that form the axons of the ON, we used a rat model of myelin oligodendrocyte glycoprotein (MOG)-induced EAE. Optic neuritis was diagnosed by recording visual evoked potentials, and RGC function was monitored by measuring electroretinograms. Methylprednisolone treatment significantly increased RGC apoptosis during MOG-EAE. By Western blot analysis, we identified the underlying molecular mechanism: a suppression of mitogen-activated protein kinase (MAPK) phosphorylation, which is a key event in an endogenous neuroprotective pathway. The methylprednisolone-induced inhibition of MAPK phosphorylation was calcium dependent. Hence, we provide evidence for negative effects of steroid treatment on neuronal survival during chronic inflammatory autoimmune disease of the CNS, which should result in a reevaluation of the current therapy regimen. PMID- 12904461 TI - Paranodal interactions regulate expression of sodium channel subtypes and provide a diffusion barrier for the node of Ranvier. AB - The node of Ranvier is a distinct domain of myelinated axons that is highly enriched in sodium channels and is critical for impulse propagation. During development, the channel subtypes expressed at the node undergo a transition from Nav1.2 to Nav1.6. Specialized junctions that form between the paranodal glial membranes and axon flank the nodes and are candidates to regulate their maturation and delineate their boundaries. To investigate these roles, we characterized node development in mice deficient in contactin-associated protein (Caspr), an integral junctional component. Paranodes in these mice lack transverse bands, a hallmark of the mature junction, and exhibit progressive disruption of axon-paranodal loop interactions in the CNS. Caspr mutant mice display significant abnormalities at central nodes; components of the nodes progressively disperse along axons, and many nodes fail to mature properly, persistently expressing Nav1.2 rather than Nav1.6. In contrast, PNS nodes are only modestly longer and, although maturation is delayed, eventually all express Nav1.6. Potassium channels are aberrantly clustered in the paranodes; these clusters are lost over time in the CNS, whereas they persist in the PNS. These findings indicate that interactions of the paranodal loops with the axon promote the transition in sodium channel subtypes at CNS nodes and provide a lateral diffusion barrier that, even in the absence of transverse bands, maintains a high concentration of components at the node and the integrity of voltage-gated channel domains. PMID- 12904462 TI - Patterned vision causes CRE-mediated gene expression in the visual cortex through PKA and ERK. AB - Normal visual experience during postnatal development is necessary for the maturation of visual cortical circuits and acts through molecular mechanisms that are still poorly understood. Recently, it has been shown that ERK (extracellular signal-regulated kinase) 1/2, protein kinase A (PKA), and CREB (cAMP response element-binding protein) are crucial factors for experience-dependent development of the visual cortex, but very little is known about the role of visual experience in their activation. Here, we show that visual stimulation after a brief period of dark rearing caused a transient ERK activation in the visual cortex. Visually induced ERK activation occurred primarily in excitatory neurons of layers II-III and VI and was prevented by binocular lid suture. ERK phosphorylation was strongly reduced by cortical infusion with the cAMP-PKA inhibitor Rp-8-Cl-cAMPS, thus establishing a link between PKA and ERK activation. To analyze the downstream consequences of ERK and PKA signaling, we studied the action of visual stimulation on transcription of genes controlled by CREB in transgenic mice carrying the LacZ reporter gene under the control of the CRE (cAMP response element) promoter. Visual stimulation triggered a prolonged episode of CRE-mediated gene expression in the visual cortex that was suppressed by infusion with the ERK inhibitor U0126. Cortical administration of Rp-8-Cl cAMPS attenuated the experience-dependent activation of CRE-mediated gene transcription. These results show that ERK phosphorylation in visual cortical neurons represents a molecular readout of patterned visual stimuli and that visual activation of ERK involves the cAMP-PKA system. Finally, because CRE mediated gene expression was totally dependent on ERK activation, we suggest that PKA action on CRE-mediated gene expression is mediated by ERK. PMID- 12904463 TI - Selective elimination of corticogeniculate feedback abolishes the electroencephalogram dependence of primary visual cortical receptive fields and reduces their spatial specificity. AB - The role of corticogeniculate feedback in the organization, function, and state dependence of visual responses and receptive fields (RFs) is not well understood. We investigated the contribution of the corticogeniculate loop to state-dependent changes of characteristics of the primary visual cortex response by using a novel approach of eliminating corticogeniculate projection neurons with targeted neuronal apoptosis. Experiments were performed in anesthetized cats (N2O plus halothane) with parallel recordings of single units from experimental (right) and control (left) hemispheres approximately 2 weeks after induction of apoptosis. Within the experimental hemispheres, neurons of area 17 and of the dorsal lateral geniculate nucleus (dLGN) showed an unusually enhanced and prolonged tonic visual response during episodes of synchronized (syn) EEG activity, whereas response levels during less synchronized states were almost normal. In addition, dLGN cells showed a reduced tendency for burst firing and a less regular spike interval distribution compared with those of controls. These changes are likely attributable to a tonic depolarization of dLGN relay neurons or, more likely, to a decreased responsiveness of thalamic inhibitory processes to cortical feedback. Cortical neurons also displayed an activity-dependent increase in RF size, in contrast to an almost activity-invariant RF size of controls, a phenomenon likely related to the elimination of collateral, intracortical projections of layer 6 neurons. Together, these results demonstrate that selective chronic elimination of corticogeniculate feedback results in the loss of EEG-correlated differences of visual processing in the remaining thalamocortical network, accompanied by a significant increase in excitability during syn EEG, at the expense of noticeably reduced spatial receptive-field specificity in the remaining cortical neurons. PMID- 12904464 TI - Synergistic effects of brain-derived neurotrophic factor and chondroitinase ABC on retinal fiber sprouting after denervation of the superior colliculus in adult rats. AB - Damage to the adult CNS often causes devastating and permanent deficits because of the limited capacity of the brain for anatomical reorganization. The finding that collateral sprouting of uninjured fiber tracts mediates recovery of function prompts the search for experimental strategies that stimulate axonal plasticity after CNS trauma. Here we characterize treatments that promote the sprouting of undamaged retinal afferents into the denervated superior colliculus (SC) after a partial retinal lesion in the adult rat. Delivery of brain-derived neurotrophic factor (BDNF) was performed to enhance the intrinsic potential of retinal ganglion cells to reelongate their axons. Reduction of the neurite growth inhibitory properties of the adult SC was accomplished via treatment with chondroitinase ABC (C-ABC), which degrades chondroitin sulfate proteoglycans. Retinal axons were labeled via intraocular injections of fluorescently tagged cholera toxin B subunit, and fiber sprouting within the denervated SC was measured by quantitative laser-scanning confocal microscopy 1 week after the retinal lesion. We found that both the administration of BDNF and the injection of C-ABC induce significant sprouting of retinal afferents into the collicular scotoma. Remarkably, the combined treatment with BDNF and C-ABC showed synergistic effects on axon growth. Colocalization analysis with anti-synapsin antibodies demonstrated synapse formation by the sprouting axons. These results suggest that the combined treatment with BDNF and C-ABC can be relevant in therapies for the repair of the damaged adult CNS. PMID- 12904465 TI - Ex vivo adenoviral vector-mediated neurotrophin gene transfer to olfactory ensheathing glia: effects on rubrospinal tract regeneration, lesion size, and functional recovery after implantation in the injured rat spinal cord. AB - The present study uniquely combines olfactory ensheathing glia (OEG) implantation with ex vivo adenoviral (AdV) vector-based neurotrophin gene therapy in an attempt to enhance regeneration after cervical spinal cord injury. Primary OEG were transduced with AdV vectors encoding rat brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3), or bacterial marker protein beta-galactosidase (LacZ) and subsequently implanted into adult Fischer rats directly after unilateral transection of the dorsolateral funiculus. Implanted animals received a total of 2 x 105 OEG that were subjected to transduction with neurotrophin encoding AdV vector, AdV-LacZ, or no vector, respectively. At 4 months after injury, lesion volumes were smaller in all OEG implanted rats and significantly reduced in size after implantation of neurotrophin-encoding AdV vector-transduced OEG. All OEG grafts were filled with neurofilament-positive axons, and AdV vector mediated expression of BDNF by implanted cells significantly enhanced regenerative sprouting of the rubrospinal tract. Behavioral analysis revealed that OEG-implanted rats displayed better locomotion during horizontal rope walking than unimplanted lesioned controls. Recovery of hind limb function was also improved after implantation of OEG that were transduced with a BDNF- or NT-3 encoding AdV vector. Hind limb performance during horizontal rope locomotion did directly correlate with lesion size, suggesting that neuroprotective effects of OEG implants contributed to the level of functional recovery. Thus, our results demonstrate that genetic engineering of OEG not only resulted in a cell that was more effective in promoting axonal outgrowth but could also lead to enhanced recovery after injury, possibly by sparing of spinal tissue. PMID- 12904466 TI - Presynaptic capacitance measurements and Ca2+ uncaging reveal submillisecond exocytosis kinetics and characterize the Ca2+ sensitivity of vesicle pool depletion at a fast CNS synapse. AB - The intracellular Ca2+ sensitivity of synaptic vesicle fusion is an important determinant of transmitter release probability, but it is unknown for most CNS synapses. We combined whole-cell membrane capacitance measurements and Ca2+ uncaging at the large calyx of Held nerve terminals to determine the Ca2+ sensitivity of synaptic vesicle fusion at a glutamatergic CNS synapse, independent of recording EPSCs. Capacitance increases measured 30-50 msec after elevating the intracellular Ca2+ concentration ([Ca2+]i) by Ca2+ uncaging were half-maximal at approximately 5 microm [Ca2+]i. At 10 microm [Ca2+]i, capacitance increases reached maximal values (256 +/- 125 fF; mean +/- SD), indicating the depletion of an average pool of approximately 4000 readily releasable vesicles. Vesicle pool depletion was confirmed in cross-depletion experiments, in which capacitance responses were measured after Ca2+ uncaging, or after combined stimuli of prolonged presynaptic depolarizations and Ca2+ uncaging. To analyze the Ca2+-dependent rates of vesicle pool depletion, the capacitance rise after Ca2+ uncaging was fitted with single- or double-exponential functions. The fast time constants of double-exponential fits, and the time constants of single exponential fits were 2-3 msec at 10-15 microm [Ca2+]i and reached submillisecond values at 30 microm [Ca2+]i. These results suggest that three to five readily releasable vesicles can fuse within <1 msec at each active zone of a calyx of Held, given that [Ca2+]i rises sufficiently high. Submillisecond kinetics of exocytosis are reached at significantly lower [Ca2+]i than at ribbon-type sensory synapses previously investigated by capacitance measurements. PMID- 12904467 TI - Selective antagonism of GluR5 kainate-receptor-mediated synaptic currents by topiramate in rat basolateral amygdala neurons. AB - Topiramate is a widely used antiepileptic agent whose mechanism of action is poorly understood. The drug has been reported to interact with various ion channel types, including AMPA/kainate receptors. In whole-cell voltage-clamp recordings from principal neurons of the rat basolateral amygdala, topiramate at low concentrations (IC50, approximately 0.5 microm) selectively inhibited pharmacologically isolated excitatory synaptic currents mediated by kainate receptors containing the GluR5 subunit. Topiramate also partially depressed predominantly AMPA-receptor-mediated EPSCs, but with lower efficacy. Topiramate did not alter the degree of facilitation in paired-pulse experiments, and it reduced the amplitude of miniature EPSCs without affecting their frequency, demonstrating that the block of synaptic responses occurs postsynaptically. Inhibition of GluR5 kainate receptors could represent a key mechanism underlying the anticonvulsant activity of topiramate. Moreover, these results support the concept that GluR5 kainate receptors represent a novel target for antiepileptic drug development. PMID- 12904468 TI - Functional integration of embryonic stem cell-derived neurons in hippocampal slice cultures. AB - The generation of neurons and glia from pluripotent embryonic stem (ES) cells represents a promising strategy for the study of CNS development and repair. ES cell-derived neural precursors have been shown to develop into morphologically mature neurons and glia when grafted into brain and spinal cord. However, there is a surprising shortage of data concerning the functional integration of ES cell derived neurons (ESNs) into the host CNS tissue. Here, we use ES cells engineered to express enhanced green fluorescent protein (EGFP) only in neuronal progeny to study the functional properties of ESNs during integration into long-term hippocampal slice cultures. After incorporation into the dentate gyrus, EGFP+ donor neurons display a gradual maturation of their intrinsic discharge behavior and a concomitant increase in the density of voltage-gated Na+ and K+ channels. Integrated ESNs express AMPA and GABA(A) receptor subunits. Most importantly, neurons derived from ES cells receive functional glutamatergic and GABAergic synapses from host neurons. Specifically, we demonstrate that host perforant path axons form synapses onto integrated ESNs. These synapses between host and ES cell derived neurons display pronounced paired-pulse facilitation indicative of intact presynaptic short-term plasticity. Thus, ES cell-derived neural precursors generate functionally active neurons capable of integrating into the brain circuitry. PMID- 12904470 TI - A broad role for melanopsin in nonvisual photoreception. AB - The rod and cone photoreceptors that mediate visual phototransduction in mammals are not required for light-induced circadian entrainment, negative masking of locomotor activity, suppression of pineal melatonin, or the pupillary light reflex. The photopigment melanopsin has recently been identified in intrinsically photosensitive retinal ganglion cells (RGCs) that project to the suprachiasmatic nucleus (SCN), intergeniculate leaflet (IGL), and olivary pretectal nucleus, suggesting that melanopsin might influence a variety of irradiance-driven responses. We have found novel projections from RGCs that express melanopsin mRNA to the ventral subparaventricular zone (vSPZ), a region involved in circadian regulation and negative masking, and the sleep-active ventrolateral preoptic nucleus (VLPO) and determined the subsets of melanopsin-expressing RGCs that project to the SCN, the pretectal area (PTA), and the IGL division of the lateral geniculate nucleus (LGN). Melanopsin was expressed in the majority of RGCs that project to the SCN, vSPZ, and VLPO and in a subpopulation of RGCs that innervate the PTA and the IGL but not in RGCs projecting to the dorsal LGN or superior colliculus. Two-thirds of RGCs containing melanopsin transcript projected to each of the SCN and contralateral PTA, and one-fifth projected to the ipsilateral IGL. Double-retrograde tracing from the SCN and PTA demonstrated a subpopulation of RGCs projecting to both sites, most of which contained melanopsin mRNA. Our results suggest that melanopsin expression defines a subset of RGCs that play a broad role in the regulation of nonvisual photoreception, providing collateralized projections that contribute to circadian entrainment, negative masking, the regulation of sleep-wake states, and the pupillary light reflex. PMID- 12904469 TI - Insulin receptor substrate-2 deficiency impairs brain growth and promotes tau phosphorylation. AB - Insulin resistance and diabetes might promote neurodegenerative disease, but a molecular link between these disorders is unknown. Many factors are responsible for brain growth, patterning, and survival, including the insulin-insulin-like growth factor (IGF)-signaling cascades that are mediated by tyrosine phosphorylation of insulin receptor substrate (IRS) proteins. Irs2 signaling mediates peripheral insulin action and pancreatic beta-cell function, and its failure causes diabetes in mice. In this study, we reveal two important roles for Irs2 signaling in the mouse brain. First, disruption of the Irs2 gene reduced neuronal proliferation during development by 50%, which dissociated brain growth from Irs1-dependent body growth. Second, neurofibrillary tangles containing phosphorylated tau accumulated in the hippocampus of old Irs2 knock-out mice, suggesting that Irs2 signaling is neuroprotective. Thus, dysregulation of the Irs2 branch of the insulin-Igf-signaling cascade reveals a molecular link between diabetes and neurodegenerative disease. PMID- 12904471 TI - Integrin requirement for hippocampal synaptic plasticity and spatial memory. AB - The establishment of memory requires coordinated signaling between presynaptic and postsynaptic terminals in the CNS. The integrins make up a large family of cell adhesion receptors that are known to mediate bidirectional signaling between cells or between cells and their external environment. We show here that many different integrins, including alpha3 and alpha5, are expressed broadly in the adult mouse brain and are associated with synapses. Mice with genetically reduced expression of alpha3 integrin fail to maintain long-term potentiation (LTP) generated in hippocampal CA1 neurons. Mice with reduced expression of the alpha3 and alpha5 integrins exhibit a defect in paired-pulse facilitation. Mice with reduced expression of alpha3, alpha5, and alpha8 are defective in hippocampal LTP and spatial memory in the water maze but have normal fear conditioning. These results demonstrate that several different integrins are involved in physiological plasticity and provide the first evidence of their requirement for behavioral plasticity in vertebrates. PMID- 12904472 TI - Disparity-based coding of three-dimensional surface orientation by macaque middle temporal neurons. AB - Gradients of binocular disparity across the visual field provide a potent cue to the three-dimensional (3-D) orientation of surfaces in a scene. Neurons selective for 3-D surface orientation defined by disparity gradients have recently been described in parietal cortex, but little is known about where and how this selectivity arises within the visual pathways. Because the middle temporal area (MT) has previously been implicated in depth perception, we tested whether MT neurons could signal the 3-D orientation (as parameterized by tilt and slant) of planar surfaces that were depicted by random-dot stereograms containing a linear gradient of horizontal disparities. We find that many MT neurons are tuned for 3 D surface orientation, and that tilt and slant generally have independent effects on MT responses. This separable coding of tilt and slant is reminiscent of the joint coding of variables in other areas (e.g., orientation and spatial frequency in V1). We show that tilt tuning remains unchanged when all coherent motion is removed from the visual stimuli, indicating that tilt selectivity is not a byproduct of 3-D velocity coding. Moreover, tilt tuning is typically insensitive to changes in the mean disparity (depth) of gradient stimuli, indicating that tilt tuning cannot be explained by conventional tuning for frontoparallel disparities. Finally, we explore the receptive field mechanisms underlying selectivity for 3-D surface orientation, and we show that tilt tuning arises through heterogeneous disparity tuning within the receptive fields of MT neurons. Our findings show that MT neurons carry high-level signals about 3-D surface structure, in addition to coding retinal image velocities. PMID- 12904473 TI - Activity-regulated dynamic behavior of early dendritic protrusions: evidence for different types of dendritic filopodia. AB - Dendritic filopodia are long and thin protrusions that occur predominantly during early development of the mammalian CNS. The function of dendritic filopodia is unknown, but they could serve to form early synapses, to generate spines, or to regulate dendritic branching and growth. We used two-photon imaging to characterize the motile behavior of dendritic protrusions during early postnatal development (P2-P12) in pyramidal neurons from acute slices of mouse neocortex. Dendritic protrusions in immature neurons are highly dynamic, and this motility is actin based. Motility and turnover of these early protrusions decreases throughout development, mirroring an increase in their average lifetime and density. Interestingly, density, motility, and length of filopodia are greater in dendritic growth cones than in dendritic shafts. These growth cones disappear after P5. Blocking synaptic transmission globally using TTX or calcium-free solutions led to a 40-120% increase in the density and length of dendritic filopodia in shafts but not in growth cones. Moreover, blocking ionotropic glutamate receptors resulted in an approximately 35% decrease in the density and turnover of shaft filopodia, whereas focal glutamate application led to a 75% increase in the length of shaft filopodia, but neither manipulation affected growth cone filopodia. Our results support the existence of two populations of filopodia, in growth cones and shafts, which are differentially regulated by neuronal activity. We propose that filopodia in dendritic growth cones are involved in dendritic growth and branching in an activity-independent manner, whereas shaft filopodia are responsible for activity-dependent synaptogenesis and, in some cases, may become dendritic spines. PMID- 12904474 TI - Transgenic mice expressing green fluorescent protein under the control of the melanocortin-4 receptor promoter. AB - The melanocortin-4 receptor (MC4-R) is an important regulator of energy homeostasis, and evidence suggests that MC4-R-expressing neurons are downstream targets of leptin action. MC4-Rs are broadly expressed in the CNS, and the distribution of MC4-R mRNA has been analyzed most extensively in the rat. However, relatively little is known concerning chemical profiles of MC4-R expressing neurons. The extent to which central melanocortins act presynaptically or postsynaptically on MC4-Rs is also unknown. To address these issues, we have generated a transgenic mouse line expressing green fluorescent protein (GFP) under the control of the MC4-R promoter, using a modified bacterial artificial chromosome. We have confirmed that the CNS distribution of GFP-producing cells is identical to that of MC4-R mRNA in wild-type mice and that nearly all GFP producing cells coexpress MC4-R mRNA. For example, cells coexpressing GFP and MC4 R mRNA were distributed in the paraventricular hypothalamic nucleus (PVH) and the dorsal motor nucleus of the vagus (DMV). MC4-R promotor-driven GFP expression was found in PVH cells producing thyrotropin-releasing hormone and in cholinergic DMV cells. Finally, we have observed that a synthetic MC3/4-R agonist, MT-II, depolarizes some GFP-expressing cells, suggesting that MC4-Rs function postsynaptically in some instances and may function presynaptically in others. These studies extend our knowledge of the distribution and function of the MC4-R. The transgenic mouse line should be useful for future studies on the role of melanocortin signaling in regulating feeding behavior and autonomic homeostasis. PMID- 12904476 TI - The statistical structure of human speech sounds predicts musical universals. AB - The similarity of musical scales and consonance judgments across human populations has no generally accepted explanation. Here we present evidence that these aspects of auditory perception arise from the statistical structure of naturally occurring periodic sound stimuli. An analysis of speech sounds, the principal source of periodic sound stimuli in the human acoustical environment, shows that the probability distribution of amplitude-frequency combinations in human utterances predicts both the structure of the chromatic scale and consonance ordering. These observations suggest that what we hear is determined by the statistical relationship between acoustical stimuli and their naturally occurring sources, rather than by the physical parameters of the stimulus per se. PMID- 12904475 TI - Substance P Acts through local circuits within the rat dorsal raphe nucleus to alter serotonergic neuronal activity. AB - Basic and clinical studies suggest that neurokinin 1 (NK1) receptor antagonists have efficacy in the treatment of affective disorders through effects on the dorsal raphe nucleus (DR), a source of forebrain-projecting serotonin (5-HT) neurons that has also been implicated in affective disorders. To investigate the regulation of the DR-5-HT system by NK1 receptors, the effects of substance P (an NK1 agonist) on rat DR neuronal activity were characterized. Most of the DR neurons (83%; n = 47 total) were inhibited by substance P microinfusion into the DR, and in some cases (17%) this was preceded by a brief activation. Pure excitation was observed in a small population of neurons (17%) that were localized in the dorsal DR, where NK1 receptors are most dense. Sendide, a selective NK1 antagonist, attenuated the effects of substance P, indicating that they were mediated by NK1 receptor activation. The selective 5-HT1A antagonist, WAY 100635, administered systemically or into the DR, prevented the inhibitory effects of substance P, implicating DR 5-HT1A receptors in this response. Finally, microinfusion of the excitatory amino acid antagonist, kynurenic acid, into the DR prevented both excitatory and inhibitory effects. The results suggest that NK1 receptor activation in the DR excites a population of 5-HT neurons via glutamatergic transmission. This results in 5-HT release throughout the DR, activation of 5-HT1A receptors, and subsequent inhibition. Interactions between NK1 and 5-HT1A receptors within DR neural networks may contribute to the mechanism of action of novel antidepressants acting at NK1 receptors. PMID- 12904478 TI - Functional changes of glial glutamate transporter GLT-1 during ischemia: an in vivo study in the hippocampal CA1 of normal mice and mutant mice lacking GLT-1. AB - Glutamate transporters remove glutamate from the extracellular space and maintain it below neurotoxic levels under normal conditions. However, the dynamics under ischemic conditions remain to be determined. In the present study, we evaluated the function of the glial glutamate transporter (GLT-1) during brain ischemia by using an in vivo brain microdialysis technique in GLT-1 mutant mice. A microdialysis probe was placed in the hippocampal CA1 of GLT-1 mutant and wild type mice, and glutamate levels were measured during 5 and 20 min ischemia. The glutamate levels in mice lacking GLT-1 were significantly higher than the corresponding glutamate levels in wild-type mice during 5 min ischemia. Delayed neuronal death was induced in the CA1 of the mice lacking GLT-1 but not in the CA1 of the wild-type mice. When ischemia was elongated to the duration of 20 min, the glutamate levels in wild-type mice were significantly higher than the corresponding glutamate levels in mice lacking GLT-1 during the last 12.5 min of 20 min ischemia. Acute neuronal death was also observed in the CA1 of wild-type mice. These results suggest that GLT-1 takes up extracellular glutamate to protect neurons in the early stage of ischemia and then releases glutamate, triggering acute neuronal death, when ischemic conditions are elongated. The function of GLT-1 may change from neuroprotective to neurodegenerative during ischemia. PMID- 12904477 TI - Neurotrophic regulation of calcium channels by the peptide neurotransmitter luteinizing hormone releasing hormone. AB - We exploited the simple organization of bullfrog paravertebral sympathetic ganglia (BFSG) to test whether the neurotransmitter peptide luteinizing hormone releasing hormone (LHRH), which generates the late slow EPSP, could also exert long-term neurotrophic control of ion channel expression. Whole-cell recordings from B-cells in BFSG showed that removal of all of the sources of ganglionic LHRH for 10 d by cutting preganglionic C-fibers in vivo caused a 28% reduction in Ca2+ current density. When BFSG B-neurons were dissociated from adult bullfrogs and maintained in a defined-medium, neuron-enriched, low-density, serum-free culture, the ICa density was increased by 49% after 6-7 d in the presence of 0.45 microm LHRH. This increase was not associated with alterations in the voltage dependence of Ca2+ current activation or inactivation and reflected a selective increase in N-type Ca2+ channel current. The increase in ICa density induced by LHRH was blocked by the transcription inhibitor actinomycin D. These results suggest that chronic exposure to a neurotransmitter that acts through G-protein-coupled receptors exerts long-term control of ion channel expression in a fully differentiated, adult sympathetic neuron in vitro or in vivo. PMID- 12904479 TI - Cytosolic prion protein in neurons. AB - Localizing the cellular prion protein (PrPC) in the brain is necessary for understanding the pathogenesis of prion diseases. However, the precise ultrastructural localization of PrPC still remains enigmatic. We performed the first quantitative study of the ultrastructural localization of PrPC in the mouse hippocampus using high-resolution cryoimmunogold electron microscopy. PrPC follows the standard biosynthetic trafficking pathway with a preferential localization in late endosomal compartments and on the plasma membrane of neurons and neuronal processes. PrPC is found with the same frequency within the synaptic specialization and perisynaptically, but is almost completely excluded from synaptic vesicles. Unexpectedly, PrP is also found in the cytosol in subpopulations of neurons in the hippocampus, neocortex, and thalamus but not the cerebellum. Cytosolic PrP may have altered susceptibility to aggregation, suggesting that these neurons might play a significant role in the pathogenesis of prion diseases, in particular those mammals harboring mutant PrP genes. PMID- 12904480 TI - Auditory cortical responses elicited in awake primates by random spectrum stimuli. AB - Contrary to findings in subcortical auditory nuclei, auditory cortex neurons have traditionally been described as spiking only at the onsets of simple sounds such as pure tones or bandpass noise and to acoustic transients in complex sounds. Furthermore, primary auditory cortex (A1) has traditionally been described as mostly tone responsive and the lateral belt area of primates as mostly noise responsive. The present study was designed to unify the study of these two cortical areas using random spectrum stimuli (RSS), a new class of parametric, wideband, stationary acoustic stimuli. We found that 60% of all neurons encountered in A1 and the lateral belt of awake marmoset monkeys (Callithrix jacchus) showed significant changes in firing rates in response to RSS. Of these, 89% showed sustained spiking in response to one or more individual RSS, a substantially greater percentage than would be expected from traditional studies, indicating that RSS are well suited for studying these two cortical areas. When firing rates elicited by RSS were used to construct linear estimates of frequency tuning for these sustained responders, the shape of the estimate function remained relatively constant throughout the stimulus interval and across the stimulus properties of mean sound level, spectral density, and spectral contrast. This finding indicates that frequency tuning computed from RSS reflects a robust estimate of the actual tuning of a neuron. Use of this estimate to predict rate responses to other RSS, however, yielded poor results, implying that auditory cortex neurons integrate information across frequency nonlinearly. No systematic difference in prediction quality between A1 and the lateral belt could be detected. PMID- 12904481 TI - Aberrant growth and differentiation of oligodendrocyte progenitors in neurofibromatosis type 1 mutants. AB - Neurofibromatosis type 1 (NF1) patients are predisposed to learning disabilities, macrocephaly, and brain tumors as well as abnormalities on magnetic resonance imaging that are postulated to result from abnormal myelination. Here we show that Nf1+/- spinal cords in adult mice have more than twofold-increased numbers of NG2+ progenitor cells. Nf1-/- embryonic spinal cords have increased numbers of Olig2+ progenitors. Also, cultures from Nf1 mutant embryos with hemizygous and biallelic Nf1 mutations have dramatically increased numbers of CNS oligodendrocyte progenitor cells. In medium that allows growth of neuroepithelial cells and glial progenitors, mutant cells hyper-respond to FGF2, have increased basal and FGF-stimulated Ras-GTP, and fail to accumulate when treated with a farnesyltransferase inhibitor. Cell accumulation results in part from increased proliferation and decreased cell death. In contrast to wild-type cells, Nf1-/- progenitors express the glial differentiation marker O4 while retaining expression of the progenitor marker nestin. Nf1 mutant progenitors also abnormally coexpress the glial differentiation markers O4 and GFAP. Importantly, Nf1-/- spinal cord-derived oligodendrocyte progenitors, which are amplified 12 fold, retain the ability to form oligodendrocytes after in vivo transplantation. The data reveal a key role for neurofibromin and Ras signaling in the maintenance of CNS progenitor cell pools and also suggest a potential role for progenitor cell defects in the CNS abnormalities of NF1 patients. PMID- 12904482 TI - Group III metabotropic glutamate receptor-mediated modulation of the striatopallidal synapse. AB - The globus pallidus (GP) is a key GABAergic nucleus in the basal ganglia (BG). The predominant input to the GP is an inhibitory striatal projection that forms the first synapse in the indirect pathway. The GP GABAergic neurons project to the subthalamic nucleus, providing an inhibitory control of these glutamatergic cells. Given its place within the BG circuit, it is not surprising that alterations in GP firing pattern are postulated to play a role in both normal and pathological motor behavior. Because the inhibitory striatal input to the GP may play an important role in shaping these firing patterns, we set out to determine the role that the group III metabotropic glutamate receptors (GluRs) play in modulating transmission at the striatopallidal synapse. In rat midbrain slices, electrical stimulation of the striatum evoked GABA(A)-mediated IPSCs recorded in all three types of GP neurons. The group III mGluR-selective agonist L-(+)-2 amino-4-phosphonobutyric acid (L-AP4) inhibited these IPSCs through a presynaptic mechanism of action. L-AP4 exhibited high potency and a pharmacological profile consistent with mediation by mGluR4. Furthermore, the effect of L-AP4 on striatopallidal transmission was absent in mGluR4 knock-out mice, providing convincing evidence that mGluR4 mediates this effect. The finding that mGluR4 may selectively modulate striatopallidal transmission raises the interesting possibility that activation of mGluR4 could decrease the excessive inhibition of the GP that has been postulated to occur in Parkinson's disease. Consistent with this, we find that intracerebroventricular injections of L-AP4 produce therapeutic benefit in both acute and chronic rodent models of Parkinson's disease. PMID- 12904484 TI - Intersegmental coordination of rhythmic motor patterns. PMID- 12904483 TI - KCNQ/M currents in sensory neurons: significance for pain therapy. AB - Neuronal hyperexcitability is a feature of epilepsy and both inflammatory and neuropathic pain. M currents [IK(M)] play a key role in regulating neuronal excitability, and mutations in neuronal KCNQ2/3 subunits, the molecular correlates of IK(M), have previously been linked to benign familial neonatal epilepsy. Here, we demonstrate that KCNQ/M channels are also present in nociceptive sensory systems. IK(M) was identified, on the basis of biophysical and pharmacological properties, in cultured neurons isolated from dorsal root ganglia (DRGs) from 17-d-old rats. Currents were inhibited by the M-channel blockers linopirdine (IC50, 2.1 microm) and XE991 (IC50, 0.26 microm) and enhanced by retigabine (10 microm). The expression of neuronal KCNQ subunits in DRG neurons was confirmed using reverse transcription-PCR and single-cell PCR analysis and by immunofluorescence. Retigabine, applied to the dorsal spinal cord, inhibited C and Adelta fiber-mediated responses of dorsal horn neurons evoked by natural or electrical afferent stimulation and the progressive "windup" discharge with repetitive stimulation in normal rats and in rats subjected to spinal nerve ligation. Retigabine also inhibited responses to intrapaw application of carrageenan in a rat model of chronic pain; this was reversed by XE991. It is suggested that IK(M) plays a key role in controlling the excitability of nociceptors and may represent a novel analgesic target. PMID- 12904485 TI - Branching thalamic afferents link action and perception. AB - Recent observations of single axons and review of older literature show that axons afferent to the thalamus commonly branch, sending one branch to the thalamus and another to a motor or premotor center of the brain stem. That is, the messages that the thalamus relays to the cerebral cortex can be regarded as copies of motor instructions. This pattern of axonal branching is reviewed, particularly for the somatosensory and the visual pathways. The extent to which this anatomical evidence relates to views that link action to perception is explored. Most pathways going through the thalamus to the cortex are already involved in motor mechanisms. These motor links occur before and during activity in the parallel and hierarchical corticocortical circuitry that currently forms the focus of many studies of perceptual processing. PMID- 12904486 TI - Task-specific internal models for kinematic transformations. AB - Numerous studies of motor learning have focused on how people adapt their reaching movements to novel dynamic and visuomotor perturbations that alter the actual or visually perceived motion of the hand. An important finding from this work is that learning of novel dynamics generalizes across different movement tasks. Thus adaptation to an unusual force field generalizes from center-out reaching movements to circular movements (Conditt et al. 1997). This suggests that subjects acquired an internal model of the dynamic environment that could be used to determine the motor commands needed for untrained movements. Using a task interference paradigm, we investigated whether transfer across tasks is also observed when learning visuomotor transformations. On day 1, all subjects adapted to a +30 degrees rotation while making center-out-and-back reaching movements. After a delay of 5 min, different groups of subjects then adapted to a -30 degrees rotation while performing either a continuous tracking task, a figure eight drawing task, or the center-out-and-back reaching task. All subjects were then retested the next day on the +30 degrees rotation in the reaching task. As expected, subjects who experienced the opposing rotations while performing the same reaching tasks showed no retention of learning for the first rotation when tested on day 2 (Krakauer et al. 1999). In contrast, such retrograde interference was not observed in the two groups of subjects who experienced the opposing rotations while performing different tasks. In fact, their performance on day 2 was similar to that of control subjects who never experienced the opposite rotation. This lack of interference suggests that memory resources for visuomotor rotations are task specific. PMID- 12904487 TI - Dopamine modulation of calcium currents in pyloric neurons of the lobster stomatogastric ganglion. AB - We examined the dopamine (DA) modulation of calcium currents (ICa) that could contribute to the plasticity of the pyloric network in the lobster stomatogastric ganglion. Pyloric somata were voltage-clamped under conditions designed to block voltage-gated Na+, K+, and H currents. Depolarizing steps from -60 mV generated voltage-dependent, inward currents that appeared to originate in electrotonically distal, imperfectly clamped regions of the cell. These currents were blocked by Cd2+ and enhanced by Ba2+ but unaffected by Ni2+. Dopamine enhanced the peak ICa in the pyloric constrictor (PY), lateral pyloric (LP), and inferior cardiac (IC) neurons and reduced peak ICa in the ventricular dilator (VD), pyloric dilator (PD), and anterior burster (AB) neurons. All of these effects, except for the AB, are consistent with DA's excitation or inhibition of firing in the pyloric neurons. Enhancement of ICa in PY and LP neurons and reduction of ICa in VD and PD neurons are also consistent with DA-induced synaptic strength changes via modulation of presynaptic ICa. However, the reduction of ICa in AB suggests that DA's enhancement of AB transmitter release is not directly mediated through presynaptic ICa. ICa in PY and PD neurons was more sensitive to nifedipine block than in AB neurons. In addition, nifedipine blocked DA's effects on ICa in the PY and PD neurons but not in the AB neuron. Thus the contribution of specific calcium channel subtypes carrying the total ICa may vary between pyloric neuron classes, and DA may act on different calcium channel subtypes in the different pyloric neurons. PMID- 12904488 TI - Analysis of recordings of single-unit firing and population activity in the dorsal subiculum of unrestrained, freely moving rats. AB - We examined neuronal activity in the dorsal subiculum of unrestrained, male adult Wistar rats, which were implanted with a moveable eight-electrode microdrive. The subiculum is the primary hippocampal formation output area and is comparatively uninvestigated neurophysiologically. We compared subicular unit activity and the subicular EEG while rats occupied a small, restricted environment and also correlated neuronal activity with the ongoing behavior of the animal. Units were separated using their electrophysiological characteristics into bursting units, regular spiking units, theta-modulated units, and fast spiking units. The bursting and regular spiking unit classes are similar to hippocampal CA1 units, whereas the fast spiking units appear to be interneurons. Bursting units were variable in their behavior: some units bursted regularly, and others bursted only occasionally. Theta-modulated units have not been described before; these were similar to regular spiking units in all respects except that they increased their firing significantly when theta oscillations were present in the simultaneous EEG record. Subicular EEG was similar to hippocampal EEG, with theta oscillations dominating "alert, moving" behaviors, while large amplitude irregular activity (LIA), which included sharp waves, predominated when theta oscillations were not present, mainly during "alert, still" and "quiet" behaviors. A relatively small proportion of subicular recordings (approximately 32%) were phase-locked to theta; this is a smaller proportion than in areas from which the subiculum takes major inputs. The relative lack of entrainment of subicular neurons by this important intrinsic rhythm is suggestive of a limit to which theta might be capable of affecting both subicular and hippocampal information processing more generally. PMID- 12904489 TI - Functional organization of parietal neuronal responses to optic-flow stimuli. AB - We analyzed the dissimilarity matrix of neuronal responses to moving visual stimuli using tree clustering and multidimensional scaling (MDS). Single-cell activity was recorded in area 7a while random dots moving coherently in eight different kinds of motion (right-, left-, up-, and downward, clockwise, counterclockwise, expansion, contraction) were presented to behaving monkeys with eyes fixated. Tree clustering analyses showed that the [rightward, leftward], [upward, downward], and [clockwise, counterclockwise]] motions were clustered in three separate branches (i.e., horizontal, vertical, and rotatory motion, respectively). In contrast, expansion was in a lone branch, whereas contraction was also separate but within a larger cluster. The distances among these clusters were then subjected to an MDS analysis to identify the dimensions underlying the tree clustering observed. This analysis revealed two major factors in operation. The first factor separated expansion from all other stimulus motions, which seems to reflect the prominence of expansion during the common activity of locomotion. In contrast, the second factor separated planar motions from motion in depth, which suggests that the latter may hold a special place in visual motion processing. PMID- 12904490 TI - Neuromotor noise limits motor performance, but not motor adaptation, in children. AB - Children do not typically appear to move with the same skill and dexterity as adults, although they can still improve their motor performance in specific tasks with practice. One possible explanation is that their motor performance is limited by an inherently higher level of movement variability, but that their motor adaptive ability is robust to this variability. To test this hypothesis, we examined motor adaptation of 43 children (ages 6-17) and 12 adults as they reached while holding the tip of a lightweight robot. The robot applied either a predictable, velocity-dependent field (the "mean field") or a similar field that incorporated stochastic variation (the "noise field"), thereby further enhancing the variability of the subjects' movements. We found that children exhibited greater initial trial-to-trial variability in their unperturbed movements but were still able to adapt comparably to adults in both the mean and noise fields. Furthermore, the youngest children (ages 6-8) were able to reduce their variability with practice to levels comparable to the remaining children groups although not as low as adults. These results indicate that children as young as age 6 possess adult-like neural systems for motor adaptation and internal model formation that allow them to adapt to novel dynamic environments as well as adults on average despite increased neuromotor or environmental noise. Performance after adaptation is still more variable than adults, however, indicating that movement inconsistency, not motor adaptation inability, ultimately limits motor performance by children and may thus account for their appearance of incoordination and more frequent motor accidents (e.g., spilling, tripping). The results of this study also suggest that movement variability in young children may arise from two sources--a relatively constant, intrinsic source related to fundamental physiological constraints of the developing motor system and a more rapidly modifiable source that is modulated depending on the current motor context. PMID- 12904491 TI - Perception of self-motion from peripheral optokinetic stimulation suppresses visual evoked responses to central stimuli. AB - In a previous functional neuroimaging study we found that early visual areas deactivated when a rotating optical flow stimulus elicited the illusion of self motion (vection) compared with when it was perceived as a moving object. Here, we investigated whether electrical cortical responses to an independent central visual probe stimulus change as a function of whether optical flow stimulation in the periphery induces the illusion of self-motion or not. Visual-evoked potentials (VEPs) were obtained in response to pattern-reversals in the central visual field in the presence of a constant peripheral large-field optokinetic stimulus that rotated around the naso-occipital axis and induced intermittent sensations of vection. As control, VEPs were also recorded during a stationary peripheral stimulus and showed no difference than those obtained during optokinetic stimulation. The VEPs during constant peripheral stimulation were then divided into two groups according to the time spans where the subjects reported object- or self-motion, respectively. The N70 VEP component showed a significant amplitude reduction when, due to the peripheral stimulus, subjects experienced self-motion compared to when the peripheral stimulus was perceived as object-motion. This finding supplements and corroborates our recent evidence from functional neuroimaging that early visual cortex deactivates when a visual flow stimulus elicits the illusion of self-motion compared with when the same sensory input is interpreted as object-motion. This dampened responsiveness might reflect a redistribution of sensorial and attentional resources when the monitoring of self-motion relies on a sustained and veridical processing of optic flow and may be compromised by other sources of visual input. PMID- 12904492 TI - Role of feedforward control of movement stability in reducing slip-related balance loss and falls among older adults. AB - Human upright posture is inherently unstable. To counter the mechanical effect of a large-scale perturbation such as a slip, the CNS can make adaptive adjustments in advance to improve the stability of the body center-of-mass (COM) state (i.e., its velocity and position). Such feedforward control relies on an accurate internal representation of stability limits, which must be a function of anatomical, physiological, and environmental constraints and thus should be computationally deducible based on physical laws of motion. We combined an empirical approach with mathematical modeling to verify the hypothesis that an adaptive improvement in feedforward control of COM stability correlated with a subsequent reduction in balance loss. Forty-one older adults experienced a slip during a sit-to-stand task in a block of slip trials, followed by a block of nonslip trials and a re-slip trial. Their feedforward control of COM stability was quantified as the shortest distance between its state measured at seat-off (slip onset) and the mathematically predicted feasible stability region boundary. With adaptation to repeated slips, older adults were able to exponentially reduce their incidence of falls and backward balance loss, attributable significantly to their improvement in feedforward control of stability. With exposure to slip and nonslip conditions, subjects began to select "optimal" movements that improved stability under both conditions, reducing the reliance on prior knowledge of forthcoming perturbations. These results can be fully accounted for when we assume that an internal representation of the COM stability limits guides the adaptive improvements in the feedforward control of stability. PMID- 12904493 TI - Activation of NMDA receptors in rat dentate gyrus granule cells by spontaneous and evoked transmitter release. AB - Activation of N-methyl-D-aspartate (NMDA) receptors by synaptically released glutamate in the nervous system is usually studied using evoked events mediated by a complex mixture of AMPA, kainate, and NMDA receptors. Here we have characterized pharmacologically isolated spontaneous NMDA receptor-mediated synaptic events and compared them to stimulus evoked excitatory postsynaptic currents (EPSCs) in the same cell to distinguish between various modes of activation of NMDA receptors. Spontaneous NMDA receptor-mediated EPSCs recorded at 34 degrees C in dentate gyrus granule cells (DGGC) have a frequency of 2.5 +/- 0.3 Hz and an average peak amplitude of 13.2 +/- 0.8 pA, a 10-90% rise time of 5.4 +/- 0.3 ms, and a decay time constant of 42.1 +/- 2.1 ms. The single-channel conductance estimated by nonstationary fluctuation analysis was 60 +/- 5 pS. The amplitudes (46.5 +/- 6.4 pA) and 10-90% rise times (18 +/- 2.3 ms) of EPSCs evoked from the entorhinal cortex/subiculum border are significantly larger than the same parameters for spontaneous events (paired t-test, P < 0.05, n = 17). Perfusion of 50 microM D(-)-2-amino-5-phosphonopentanoic acid blocked all spontaneous activity and caused a significant baseline current shift of 18.8 +/- 3.0 pA, thus identifying a tonic conductance mediated by NMDA receptors. The NR2B antagonist ifenprodil (10 microM) significantly reduced the frequency of spontaneous events but had no effect on their kinetics or on the baseline current or variance. At the same time, the peak current and charge of stimulus-evoked events were significantly diminished by ifenprodil. Thus spontaneous NMDA receptor-mediated events in DGGC are predominantly mediated by NR2A or possibly NR2A/NR2B receptors while the activation of NR2B receptors reduces the excitability of entorhinal afferents either directly or through an effect on the entorhinal cells. PMID- 12904494 TI - Excitatory inputs to CA1 interneurons show selective synaptic dynamics. AB - The dynamic properties of synapses between neurons in the hippocampal CA1 area are important for the frequency-dependent signal transfer of the network. We have examined the synaptic dynamics of excitatory inputs to CA1 interneurons and pyramidal cells using whole cell voltage-clamp recordings. The CA1 network was activated using extracellular stimulation electrodes at the Schaffer collaterals (feedforward activation) or at the Alveus (activation of the feedback loop). The dynamic properties of input from the Schaffer collaterals to CA1 interneurons (basket and bistratified cells) were different from the synaptic dynamics of input from the Alveus. Synaptic input from the Schaffer collaterals to CA1 interneurons showed facilitation for most frequencies. After 10 stimuli the synaptic response reached a plateau level that was approximately 150% of the first response in the train. In contrast, the plateau levels of Alveus inputs to interneurons were not different from the first responses for frequencies 11,000 ES cell clones. Although most of the gene-trap vector insertions occurred randomly throughout the genome, we found both vector independent and vector-specific integration "hot spots." Because >50% of the hot spots were vector-specific, we conclude that the most effective way to saturate the mouse genome with gene-trap insertions is by using a combination of gene-trap vectors. When a random sample of gene-trap integrations was passaged to the germ line, 59% (17 of 29) produced an observable phenotype in transgenic mice, a frequency similar to that achieved by conventional gene targeting. Thus, gene trapping allows a large-scale and cost-effective production of ES cell clones with mutations distributed throughout the genome, a resource likely to accelerate genome annotation and the in vivo modeling of human disease. PMID- 12904584 TI - Attenuation of FLOWERING LOCUS C activity as a mechanism for the evolution of summer-annual flowering behavior in Arabidopsis. AB - Plant species have evolved a wide variety of flowering habits, each adapted to maximize reproductive success in their local environment. Even within a species, accessions from different environments can exhibit markedly different flowering behavior. In Arabidopsis, some accessions are rapid-cycling summer annuals, whereas others accessions are late flowering and vernalization responsive and thus behave as winter annuals. Two genes, FLOWERING LOCUS C (FLC) and FRIGIDA (FRI), interact synergistically to confer the winter-annual habit. Previous work has shown that many summer-annual accessions contain null mutations in the FRI gene; thus it appears that these summer-annual accessions have arisen from winter annual ancestors by losing FRI function. In this work we demonstrate that naturally occurring allelic variation in FLC has provided another route to the evolution of summer-annual flowering behavior in Arabidopsis. We have identified two summer-annual accessions, Da (1)-12 and Shakhdara, that contain functional alleles of FRI, but are early flowering because of weak alleles of FLC. We have also determined that the weak allele of FLC found in Landsberg erecta is naturally occurring. Unlike accessions that have arisen because of loss-of function mutations in FRI, the FLC alleles from Da (1)-12, Shakhdara, and Landsberg erecta are not nulls; however, they exhibit lower steady-state mRNA levels than strong alleles of FLC. Sequence analysis indicates that these weak alleles of FLC have arisen independently at least twice during the course of evolution. PMID- 12904586 TI - DHA in pregnancy benefits child development. PMID- 12904585 TI - Developmental amnesia: effect of age at injury. AB - Hypoxic-ischemic events sustained within the first year of life can result in developmental amnesia, a disorder characterized by markedly impaired episodic memory and relatively preserved semantic memory, in association with medial temporal pathology that appears to be restricted to the hippocampus. Here we compared children who had hypoxic-ischemic events before 1 year of age (early group, n = 6) with others who showed memory problems after suffering hypoxic ischemic events between the ages of 6 and 14 years (late group, n = 5). Morphometric analyses of the whole brain revealed that, compared with age-matched controls, both groups had bilateral abnormalities in the hippocampus, putamen, and posterior thalamus, as well as in the right retrosplenial cortex. The two groups also showed similar reductions (approximately 40%) in hippocampal volumes. Neuropsychologically, the only significant differences between the two were on a few tests of immediate memory, where the early group surpassed the late group. The latter measures provided the only clear indication that very early injury can lead to greater functional sparing than injury acquired later in childhood, due perhaps to the greater plasticity of the infant brain. On measures of long-term memory, by contrast, the two groups had highly similar profiles, both showing roughly equivalent preservation of semantic memory combined with marked impairment in episodic memory. It thus appears that, if this selective memory disorder is a special syndrome related to the early occurrence of hypoxia-induced damage, then the effective age at injury for this syndrome extends from birth to puberty. PMID- 12904587 TI - Peanut allergy: recent advances. PMID- 12904588 TI - Postural adjustments in preterm infants at 4 and 6 months post-term during voluntary reaching in supine position. AB - Gradually it is getting clear that motor development - in particular balance control - in so-called "low-risk" preterm infants often differs from that in full term infants. However, little is known on the etiology and pathophysiology of these problems. The aim of this study was to evaluate postural behavior during reaching by means of kinetic and kinematic measurements. Preterm infants (n = 32) without cerebral palsy were investigated longitudinally at the corrected ages of 4 and 6 mo. Thirteen age-matched full-term infants served as controls. Cognitive and motor development were assessed by means of the quality of General Movements (GMs) at 4 mo and Bayley scales at 6 and 12 mo. The infants were lying supine on a forceplate reaching for a toy and the kinetics of the total body's Center of Pressure (COP) was measured in cranial-caudal and medial-lateral direction. The analysis focused on COP displacement, Vmax and oscillatory changes of the COP displacement during reaching. The kinematic analysis of reaching focused on movement units, Vmax and a compound kinematic variable reflecting the quality of reaching. The results showed that preterm infants showed a remarkable "still" postural behavior, which differed significantly from the mobile COP behavior of the full-term infants. More "still" postural behavior at 6 mo was associated with a better quality of reaching movements and with normal GMs at 4 mo. We concluded that "still" postural behavior is an adequate postural strategy of preterm infants. But it might be that this postural behavior is an indicator of later dysfunction. PMID- 12904589 TI - CBF reactivity in hypotensive and normotensive preterm infants. AB - Perinatal distress in the preterm neonate, and the consequent loss of cerebrovascular autoregulation, has been implicated in the pathogenesis of neonatal cerebral lesions. A component of this distress is thought to be hypotension. We examined the autoregulatory capacity of hypotensive and normotensive infants using the 133Xe technique to measure cerebral blood flow. Global CBF was measured during only normotension in 5 infants, and during both hypotension and normotension in 11 infants. All the infants were ventilated and blood pressure was measured using an intra-arterial catheter. Fourteen CBF measurements were made on the normotensive infants. Forty-seven CBF measurements were made on the hypotensive infants, 34 measurements during hypotension and 13 during normotension. The global CBF of the normotensive and hypotensive infants were 13.3 and 13.6 mL/100 g/min, respectively. The mean arterial blood pressure (MABP)-CBF reactivity (95% CI) of the normotensive and hypotensive infants were 1.9% (-0.8% to 4.7%)/mm Hg and 1.9% (0.8% to 3.0%)/mm Hg, respectively. The CO2 CBF reactivity (95%CI) of the normotensive and hypotensive infants was 11.1% (6.8% to 15.5%)/KPa deltaPaCO2 and 4.1% (-5.0% to 14.1%)/KPa deltaPaCO2. The implications of these calculated CBF reactivities is that normotensive infants may have intact autoregulation but with a diminished response to fluctuations in PaCO2. The hypotensive infants appear to have attenuated or absent autoregulation with little or no response in CBF to changes in PaCO2. PMID- 12904590 TI - Angiotensin II type 1 receptor A1166C polymorphism and prophylactic indomethacin treatment induced ductus arteriosus closure in very low birth weight neonates. AB - Altered pulmonary vascular resistance might be a factor for delayed closure of the ductus arteriosus (DA) in preterm infants. Angiotensin II plays a central role in the elevation of pulmonary vascular resistance. Angiotensin II exerts its vasoconstrictor effect on the angiotensin II type 1 receptor (AT1R). Homozygous carriers of the AT1R A1166C genetic variant present an exaggerated vasoconstrictor response to angiotensin II. We have investigated whether the presence of AT1R CC1166 influences the effect of prophylactic indomethacin treatment on the closure of DA until the fifth postnatal day in preterm infants. In this retrospective study detailed medical history of the first postnatal week was obtained in 159 infants born before the 33rd gestational week. All were treated by prophylactic indomethacin to induce permanent closure of the DA. On the sixth postnatal day the DA was still open in 56, whereas it was permanently closed in 103. The AT1R A1166C genotype of the infants was determined from Guthrie spots. Stepwise binary logistic regression analysis was used to assess the effect of medical conditions and genotype on the risk of patent DA (PDA). Birth weight, infantile respiratory distress, and severe hypotension were independent risk factors for PDA (p < 0.01, p < 0.05, p < 0.05, respectively). The carrier state of AT1R CC1166 was protective against PDA (p < 0.05; odds ratio, 0.067). AT1R AC1166 genotype was not associated with PDA. Our results indicate that the risk of PDA might be lower in infants of AT1R CC1166 than in those with AC or AA genotypes. PMID- 12904591 TI - Cerebral (31)P magnetic resonance spectroscopy and systemic acid-base balance during hypoxia in fetal sheep. AB - The purpose of this study was to investigate cerebral energy metabolism and acid base homeostasis during impaired oxygen supply in fetal sheep. Systemic acid-base balance was correlated with the sequence in changes of cerebral phosphorus metabolite ratios and intracellular pH. Phosphorus magnetic resonance spectra were obtained from the brain of six fetal sheep simultaneously with repeated measurements of fetal arterial oxygen saturation and acid-base balance. Fetal hypoxia was induced by gradually reducing the oxygen supply to the anesthetized pregnant ewe to establish an intended arterial pH of 7.00 or lower. The ratio of phosphocreatine to inorganic phosphate decreased from 1.08 +/- 0.10 (SD) during the control period to 0.77 +/- 0.29 at an arterial pH between 7.20 and 7.25. The inorganic phosphate level became significantly increased at an arterial pH between 7.10 and 7.15 compared with control values. With ongoing arterial acidosis, cerebral intracellular pH decreased linearly with the arterial pH. At an arterial pH of 7.00, cerebral intracellular pH was decreased from 7.18 +/- 0.03 to 6.71 +/- 0.28, and phosphocreatine and nucleoside triphosphates levels were decreased significantly. In fetal sheep brain, cerebral oxidative phosphorylation (ratio of phosphocreatine to inorganic phosphate) is already affected at a mild arterial acidosis. At an arterial pH of 7.00 or lower, nucleoside triphosphates disappeared, which almost inevitably was followed by death in fetal sheep. PMID- 12904593 TI - Activin a plasma levels at birth: an index of fetal hypoxia in preterm newborn. AB - Activin-A is a growth factor involved in cell growth and differentiation, neuronal survival, early embryonic development and erythropoiesis. Hypoxemia is a specific trigger for increasing activin-A in fetal lamb circulation. We tested the hypothesis that fetal hypoxia induces activin-A secretion in preterm newborn infants. Fifty newborn infants with gestational ages ranging from 26 to 36 wk were enrolled in a prospective study performed at the Pediatrics, Obstetrics and Reproductive Medicine Department, University of Siena, Italy. Heparinized blood samples were obtained from the umbilical vein after cord clamping, immediately after delivery. Activin A, hypoxanthine (Hx), xanthine (Xa) plasma levels and absolute nucleated red blood cell (NRBC) count were measured. Activin-A levels (p < 0.0001) and NRBC (p < 0.0001) were significantly higher in hypoxic than in non hypoxic preterm newborns. Cord activin A levels were significantly related with Hx (taua=0.64, taub=0.64, p < 0.0001) and Xa (taua=0.56, taub=0.57, p < 0.0001) levels, NRBC ((taua=-0.45, taub=-0.46, p < 0.0001) count; pH (taua=-0.47, taub= 0.48, p < 0.0001) and base deficit (taua=-0.36, taub=0.-0.36, p = 0.0002). Preterm newborns with signs of perinatal hypoxia at birth have increased activin A levels, suggesting that activin-A may reflect indirectly intrauterine hypoxia. PMID- 12904592 TI - Pulmonary surfactant protein A, B, and C mRNA and protein expression in the nitrofen-induced congenital diaphragmatic hernia rat model. AB - Neonates with congenital diaphragmatic hernia (CDH) suffer from a diaphragmatic defect, lung hypoplasia, and pulmonary hypertension, with poor lung function forming the major clinical challenge. Despite prenatal diagnosis and advanced postnatal treatment strategies, the mortality rate of CDH is still high. CDH has been subject of extensive research over the past decades, but its etiology remains unknown. A major problem with CDH is the failure to predict the individual response to treatment modalities like high-frequency ventilation, inhaled nitric oxide, and extracorporeal membrane oxygenation. In this study, we tested the possibility that CDH lungs are surfactant protein deficient, which could explain the respiratory failure and difficulties in treating CDH infants. We investigated this hypothesis in the nitrofen-induced CDH rat model and assessed the cellular concentrations of surfactant protein (SP)-A, -B, and -C mRNA with a quantitative radioactive in situ hybridization technique. No differences were observed between control and CDH lungs for SP mRNA expression patterns. The cellular concentration (mean OD) of SP-A and SP-B mRNA was similar at all stages whereas the mean OD of SP-C mRNA and the volume fraction of cells (% Area) expressing SP mRNA was higher in CDH lungs at term. Immunohistochemical analysis revealed no differences between control and CDH lungs for SP protein expression. No differences in the mean OD or % Area for the SP mRNAs were found between the ipsi- and contralateral side of CDH lungs. We conclude that there is no primary deficiency of surfactant proteins in the nitrofen-induced CDH rat model. PMID- 12904594 TI - Developmental expression of endothelial nitric oxide synthase (eNOS) in the rat liver. AB - Transition from fetal to postnatal life requires significant changes in cardiac, pulmonary, and hepatic blood flow. As such, there must be changes in vascular control in these vascular systems. Vascular resistance, a major contributor to blood flow, is mediated in the ductus arteriosus and pulmonary vasculature by endothelial nitric oxide synthase (eNOS). This study was conducted to determine the ontogeny of hepatic eNOS expression and activity. Additionally, the expression and activity of inducible nitric oxide synthase (iNOS) was measured to determine whether perinatal hypoxia resulted in detectable levels. NOS mRNA and proteins were determined by reverse transcription PCR assay and semiquantitative Western blot analysis, respectively. NOS activity was measured by the formation of [14C]-citrulline from [14C]-arginine. Localization of eNOS within the liver was determined by immunohistochemistry. eNOS mRNA was detectable at low levels at 18-d gestation and increased after birth, reaching a maximum level (4.5-fold increase) at 20 d of life. Parallel patterns for eNOS protein and activity were seen, with 6.9-fold and 16.1-fold increases, respectively. In the prenatal rat, eNOS was localized to areas of extramedullary hematopoiesis, with little signal in the sinusoids. Postnatally, there was a decrease in staining in the hematopoietic cells and a gradual increase in the staining of the endothelium of the sinusoids and central veins. iNOS mRNA and protein could not be detected at any age. eNOS expression and activity are developmentally regulated, increasing after birth coincident with an initial relative hypoxia and an increase in shear forces upon closure of the ductus venosus. PMID- 12904595 TI - Colonization of gnotobiotic mice with human gut microflora at birth protects against Escherichia coli heat-labile enterotoxin-mediated abrogation of oral tolerance. AB - Previous work has shown that the indigenous gut microflora in mice plays a protective role against Escherichia coli heat-labile enterotoxin (LT)-mediated abrogation of oral tolerance to an unrelated co-ingested protein. To assess potential protection by human gut microflora, we studied the effect of human gut microflora in a murine model. Oral tolerance was studied in adult gnotobiotic mice (i.e. ex-germ-free mice) colonized with the entire human fecal microflora and orally administered once with LT and ovalbumin. Systemic suppression of IgG, IgG1, IgG2a, and IgE antibody responses was assessed by ELISA. Both specific IgG subclasses and IgE hyporesponsiveness was induced in LT + ovalbumin-fed gnotobiotic mice, indicating that the human gut microflora can protect against the LT-mediated abrogation of oral tolerance. However, as confirmed with mouse gut microflora, this protective effect only occurs when the gut microflora is associated from birth on. Colonization of germ-free mice with a single bacterial strain, E. coli, predominant in the human and mouse gut microflora in the neonatal period, showed that this strain alone did not induce protection. These results supported the hypothesis that the natural establishment of the gut microflora in neonates crucially influenced resistance to LT-mediated abrogation of oral tolerance by reinforcing suppression of both T helper type 1- and T helper type 2-controlled responses, and suggested that sequential bacterial colonization of the gut rather than a single bacterial species may be involved in this phenomenon. PMID- 12904596 TI - Telomerase activity in cell lines of pediatric soft tissue sarcomas. AB - Telomeres and their maintenance by telomerase have been implicated to play an important role in carcinogenesis. As almost all malignant tumors express telomerase (in contrast to normal somatic cells), assessment of its activity has been proposed as a diagnostic and prognostic tool. To test the prognostic value of telomerase in pediatric soft tissue sarcoma (STS), we analyzed telomere length (by telomere restriction fragment analysis), telomerase activity (by modified telomerase repeat amplification protocol assay), and expression of human telomerase reverse transcriptase (hTERT) mRNA (by TaqMan technique) in cell lines of different types of STS from 12 children and adolescents. Telomere length (3.7 9.0 kb) showed a very heterogeneous pattern, independent of subtype of STS or the age of the patients, and it was not associated with expression of hTERT mRNA. In contrast, there was a trend of an association between hTERT and telomerase activity. The three tested cell lines of embryonal rhabdomyosarcomas demonstrated no or low (n = 2) telomerase activity, which was confirmed in two cases by a very low expression of hTERT mRNA. Thus, we suggest that the significant difference (p < 0.01) in the less aggressive clinical behavior of embryonal rhabdomyosarcomas in comparison to other subtypes may be due to differences in telomerase expression. Taken together, our cell line experiments imply that telomerase activity might be a biologic marker for stratification between STS with different clinical prognosis. PMID- 12904597 TI - Impact of intravenous antibiotic therapy on total daily energy expenditure and physical activity in cystic fibrosis children with Pseudomonas aeruginosa pulmonary exacerbation. AB - Resting energy expenditure (REE) increases during pulmonary exacerbation by Pseudomonas aeruginosa in cystic fibrosis (CF) patients, and decreases after i.v. anti-Pseudomonas aeruginosa antibiotic therapy (IVAT). However, the impact of IVAT on total energy expenditure (TEE) is unknown. The aim of this study was to assess the changes in TEE and its main components after IVAT administered at home. Body composition measured by skinfold thickness and bio-impedance analysis, energy intake (EI) assessed by a weekly diary, REE measured by indirect calorimetry (IC), TEE assessed by a technique using 24-h heart-rate monitoring method and physical activity (PA) monitored using an activity diary (AD) were assessed in 16 patients (9 boys and 7 girls) aged 12.1 +/- 2.3 y (range, 7.1-14.6 y), before and after 28 +/- 4 d including a 14-d IVAT course. After IVAT, weight increased significantly by 1.9% (32.1 +/- 7.5 versus 32.7 +/- 7.6 kg; p < 0.05), while fat mass and fat free mass increased non significantly. EI increased by 4.6% (10,797 +/- 3039 versus 11320 +/- 3074 kJ/d; p < 0.05). TEE was not affected by IVAT (7014 +/- 1929 versus 7081 +/- 1478 kJ/d) whereas REE decreased by 4.1% (5295 +/- 909 versus 5093 +/- 837 kJ/d; p < 0.05), resulting in 9.3% increase in PA assessed by AD converted to metabolic equivalent tasks (MET) (37.0 +/- 3.1 versus 40.7 +/- 4.5 MET; p < 0.05). The improvement in nutritional status after IVAT is not related to a decrease in TEE, but probably to an increase in EI and a decrease of REE after IVAT. After IVAT, the reduction in REE is probably compensated by an increase in PA in CF patients. PMID- 12904598 TI - Human cord blood leukocyte innate immune responses to defense collagens. AB - The innate immune system provides critical protection during initial infections before the generation of an appropriate adaptive (antibody or T cell mediated) immune response. These early defense mechanisms may be particularly critical for neonates in whom the adaptive immune system is not fully operational. Pattern recognition molecules target potential pathogens for destruction by the innate immune system, and likely facilitate the initiation of a pathogen-specific immune response. Defense collagens, such as C1q, MBL and SPA, comprise a family of such proteins that, via specific interactions with phagocytic cells, play a role in this first line of defense. To begin to assess the importance of these innate defense mechanisms in neonates, cord blood plasma and leukocytes were isolated, and responses to these components of the innate defense system were assessed. C1q enhanced the phagocytosis of targets suboptimally opsonized with either IgG or complement components, and this enhancement of phagocytosis was blocked by anti CD93/C1qRP MAb by 57% to 68%. Flow cytometric analysis demonstrated that neonatal monocytes and neutrophils expressed CD93/C1qRP similarly to adult cells, with several-fold greater expression on monocytes than on neutrophils and essentially no expression on lymphocytes. Superoxide production in response to multivalent C1q by neonatal neutrophils was also comparable to adult cells. We also confirm that C1q and MBL are present in neonate circulation. Thus, the data demonstrate that these recognition and effector mechanisms of the innate system are functional in the newborn and similar to that of adult cells. PMID- 12904599 TI - Control of brain intracellular bilirubin levels. PMID- 12904600 TI - Genetic regulation of branching morphogenesis: lessons learned from loss-of function phenotypes. AB - Branching morphogenesis, defined as growth and branching of epithelial tubules during embryogenesis, is a fundamental feature of renal, lung, mammary gland, submandibular gland, and pancreatic morphogenesis in mammals. Disruption of branching morphogenesis has been demonstrated to result in maldevelopment of some of these organs. Genetic studies performed in affected humans and mutant mice have implicated transcription factors, secreted growth factors, and cell surface signaling molecules as critical regulators of branching morphogenesis. These factors function within networks that appear to exert tight control over the number and location of branches. This review summarizes current knowledge regarding the molecular control of branching morphogenesis in vivo with particular emphasis on the genetic contribution to perturbed branching morphogenesis in mice and humans. PMID- 12904601 TI - Administration of drugs known to inhibit P-glycoprotein increases brain bilirubin and alters the regional distribution of bilirubin in rat brain. AB - P-glycoprotein (P-gp) is an ATP-dependent integral plasma membrane efflux pump, expressed in abundance in brain capillary endothelial cells and astrocytes. P-gp contributes to the blood-brain barrier in limiting the influx and retention of a variety of lipophilic compounds, including unconjugated bilirubin. Several drugs block P-gp function and thereby enhance intracellular accumulation of P-gp substrates. In this study we proposed that pretreatment with drugs known to inhibit P-gp function in clinically relevant doses would alter the uptake of bilirubin in the brain of 32- to 36-d-old rats. In the first arm of the study, the animals received pretreatment with an i.v. infusion of either propanolol, erythromycin, verapamil, ceftriaxone, rifampin, or saline, 10 min before an i.v. bolus of 50 mg/kg bilirubin was given. Except for the erythromycin-treated rats, all treatment groups had significantly higher brain-to-serum bilirubin ratios than control animals (p < 0.05, Welch's t test). In the second arm of the study, treatment with either ceftriaxone or rifampin or saline i.v. preceded a 50 mg/kg i.v. bolus of radioactive bilirubin. Analysis of seven different brain regions by scintillation counting showed that the distribution patterns differed significantly between the study groups (p < 0.001, ANOVA), however, not in accordance with a kernicteric staining pattern. Because of limited knowledge of expression and function of P-gp and other membrane transport proteins in the newborn, the implications of this study remain to be seen. We speculate that drugs known to inhibit P-gp function may increase the risk of bilirubin encephalopathy in the hyperbilirubinemic infant. PMID- 12904602 TI - TRAIL/Apo2L ligands induce apoptosis in malignant rhabdoid tumor cell lines. AB - Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL/Apo2L) is a potent inducer of apoptosis in various cancer cells, whereas normal cells are not sensitive to TRAIL-mediated apoptosis. Four TRAIL/Apo2L receptors (DR4, DR5, DcR1, and DcR2) have been identified. DR4 and DR5 have a death domain, whereas DcR1 and DcR2 are called decoy receptors because of their incomplete or lack of a death domain. Malignant rhabdoid tumor (MRT) is an aggressive neoplasm showing a poor prognosis because of its resistance to chemotherapeutic agents. In this study, we examined whether TRAIL could induce apoptotic cell death in MRT cell lines. We found that although half of the MRT cell lines examined were sensitive to TRAIL/Apo2L, Western blot analysis revealed that the expression of DcR2 was low in TRAIL-sensitive MRT cells. We examined the effect of doxorubicin on the expression levels of TRAIL receptors and its enhancement on the susceptibility of MRT cell lines to TRAIL. Western blot and flow cytometric analyses revealed that doxorubicin significantly increased the expression of DR5, and somewhat up regulated the expression of DR4 and DcR2. Moreover, doxorubicin, NF-kappaB inhibitor (SN50), and PI3-kinase/Akt inhibitor (wortmannin, LY294002) enhanced the susceptibility of MRT cell lines to TRAIL/Apo2L-induced apoptosis. These results suggest that TRAIL/Apo2L may provide the basis for clinical trials of TRAIL-based treatment to improve the outcome of MRT patients. PMID- 12904603 TI - L-carnitine reduces brain injury after hypoxia-ischemia in newborn rats. AB - Perinatal hypoxia-ischemia remains a significant cause of neonatal mortality and neurodevelopmental disability. Numerous lines of evidence indicate that cerebral ischemic insults disrupt normal respiratory activity in mitochondria. Carnitine (3-hydroxy-4-N-trimethylammonium-butyrate) has an essential role in fatty acid transport in the mitochondrion and in modulating potentially toxic acyl-CoA levels in the mitochondrial matrix. There are no naturally occurring esterases available to reduce the accumulation of acyl-CoA but this process can be overcome by exogenous carnitine. We used a newborn rat model of perinatal hypoxia-ischemia to test the hypothesis that treatment with l-carnitine would reduce the neuropathologic injury resulting from hypoxia-ischemia in the developing brain. We found that treatment with l-carnitine during hypoxia-ischemia reduces neurologic injury in the immature rat after both a 7- and 28-d recovery period. We saw no neuroprotective effect when l-carnitine was administered after hypoxia ischemia. Treatment with d-carnitine resulted in an increase in mortality during hypoxia-ischemia. Carnitine is easy to administer, has low toxicity, and is routinely used in neonates as well as children with epilepsy, cardiomyopathy, and inborn errors of metabolism. l-Carnitine merits further investigation as a treatment modality for the asphyxiated newborn or as prophylaxis for the at-risk fetus or newborn. PMID- 12904604 TI - Pulmonary surfactant disaturated-phosphatidylcholine (DSPC) turnover and pool size in newborn infants with congenital diaphragmatic hernia (CDH). AB - In animal CDH models, surfactant deficiency contributes to the pathophysiology of the condition but information on human disease is very limited. The aim of our study was to investigate surfactant kinetics in CDH newborns. We studied surfactant disaturated-phosphatidylcholine (DSPC) half-life, turnover and apparent pool size by stable isotope methodology in CDH newborns with no ExtraCorporeal Membrane Oxygenation (ECMO) support (n = 13, birth weight (BW) 3.2 +/- 2.2 kg, gestational age (GA) 39 +/- 0.4 wks, postnatal age 43 +/- 11 h) and in 8 term infants with no lung disease (CONTROLS, BW 2.7 +/- 0 kg, GA 38 +/- 0.8 wks, postnatal age 96 +/- 26 h). We administered a trace dose of 13C-palmitic acid dipalmitoyl-phosphatidylcholine (DPPC) through the endotracheal (ET) tube and we measured DSPC kinetics by gas chromatography-mass spectrometry from DSPC13C-enrichment decay curves obtained from sequential tracheal aspirates. DSPC amount from tracheal aspirates (TA-DSPC) was measured by gas chromatography. In CDH infants DSPC half-life was shorter (24 +/- 4 and 53 +/- 11 h, p = 0.01), turnover faster (0.6 +/- 0.1 and 1.5 +/- 0.3 d-1 p = 0.01), apparent pool size smaller (34 +/- 6 and 57 +/- 7 mg/kg body weight, p = 0.02) and tracheal aspirates DSPC amount lower (2.4 +/- 0.4 and 4.6 +/- 0.5 mg/mL Epithelial Lining Fluid (ELF), p = 0.007) than in CONTROLS. In conclusion surfactant kinetics is grossly abnormal in mechanically ventilated CDH. Whether alterations of DSPC kinetics in CDH infants are caused by a primary surfactant deficiency or are secondary to oxygen therapy and ventilator support has still to be determined. PMID- 12904605 TI - The de novo Q167K mutation in the POU1F1 gene leads to combined pituitary hormone deficiency in an Italian patient. AB - The POU1F1 gene encodes a transcription factor that is important for the development and differentiation of the cells producing GH, prolactin, and TSH in the anterior pituitary gland. Patients with POU1F1 mutations show a combined pituitary hormone deficiency with low or absent levels of GH, prolactin, and TSH. Fourteen mutations have been reported in the POU1F1 gene up to now. These genetic lesions can be inherited either in an autosomal dominant or an autosomal recessive mode. We report on the first Italian patient, a girl, affected by combined pituitary hormone deficiency. The patient was found to be positive for congenital hypothyroidism (with low TSH levels) at neonatal screening. Substitutive therapy was started, but subsequent growth was very poor, although psychomotor development was substantially normal. Hospitalized at 10 mo she showed hypotonic crises, growth retardation, delayed bone age, and facial dysmorphism. In addition to congenital hypothyroidism, GH and prolactin deficiencies were found. Mutation DNA analysis of the patient's POU1F1 gene identified the novel Q167K amino acid change at the heterozygous level. The highly conserved Q167 residue is located in the POU-specific domain. No mutation was detected in the other allele. DNA analysis in the proband's parents did not identify this amino acid substitution, suggesting a de novo genetic lesion. From these data it can be hypothesized that the Q167K mutation has a dominant negative effect. PMID- 12904606 TI - Cardiac and ocular pathologies in a mouse model of mucopolysaccharidosis type VI. AB - Mucopolysaccharidosis type VI (MPS VI) is a lysosomal storage disease caused by a deficiency of arylsulfatase B (ASB) which has its function in the sequential degradation of glycosaminoglycans (GAG). Targeted disruption of the ASB gene resulted in a mouse model of MPS VI that has been closely investigated for skeletal and chondral dysplasia. As ocular and cardiac impairment are also clinically important manifestations of the MPS VI syndrome, the present study was initiated for detailed biochemical, histologic and functional analysis of cornea, optic nerve and heart in ASB-deficient mice. Biochemical evidence for GAG-storage could be obtained for liver, kidney, spleen and myocardium as well as for heart valves, cornea and optic nerve from ASB-deficient mice. In MPS VI mice, histology revealed structural impairment of corneal stroma and epithelium as well as a thickening of the heart valves. According to histologic investigations, the optic nerve appeared not to be altered. However, GAG-storage in the dura mater could be demonstrated in MPS VI mice. Heart function was assessed by echocardiography. While the dimensions of MPS VI hearts were not altered, these hearts clearly showed decreased myocardial contraction and a 50% reduction of cardiac output. In addition, insufficiencies in the mitral and aortic valves were detected. Thus, ASB-deficient mice resemble the phenotype of human MPS VI not only in the skeletal but also in the ocular and cardiac symptoms. To our knowledge, these in vivo evaluations of heart function represent the first respective investigation of a MPS VI animal model and should provide a valuable measure for therapy studies in the MPS VI mouse. PMID- 12904607 TI - Preterm children have disturbances of white matter at 11 years of age as shown by diffusion tensor imaging. AB - Preterm birth frequently involves white matter injury and affects long-term neurologic and cognitive outcomes. Diffusion tensor imaging has been used to show that the white matter microstructure of newborn, preterm children is compromised in a regionally specific manner. However, until now it was not clear whether these lesions would persist and be detectible on long-term follow-up. Hence, we collected diffusion tensor imaging data on a 1.5-T scanner, and computed fractional anisotropy and coherence measures to compare the white matter integrity of children born preterm to that of control subjects. The subjects for the preterm group (10.9 +/- 0.29 y; n = 9; birth weight or= 2500; gestational age, >or= 37 wk). We found that the preterm group had lower fractional anisotropy values in the posterior corpus callosum and bilaterally in the internal capsules. In the posterior corpus callosum this difference in fractional anisotropy values may partially be related to a difference in white matter volume between the groups. An analysis of the coherence measure failed to indicate a group difference in the axonal organization. These results are in agreement with previous diffusion tensor imaging findings in newborn preterm children, and indicate that ex-preterm children with attention deficits have white matter disturbances that are not compensated for or repaired before 11 y of age. PMID- 12904608 TI - Needlescopic operation: surgery of the future? PMID- 12904610 TI - Dexfenfluramine and heart-valve regurgitation in Chinese patients with type 2 diabetes. AB - OBJECTIVE: To assess whether valvular lesions are associated with the use of dexfenfluramine in Chinese patients with type 2 diabetes. DESIGN: Case-control study. PATIENTS AND METHODS: Thirty-six obese Chinese patients with type 2 diabetes and a history of dexfenfluramine use during the period January 1992 and September 1997 were recruited into the study, while another 43 age- and sex matched Chinese patients with type 2 diabetes were recruited as controls. The mean age for the cases was 44.1 years (standard deviation, 11.2 years; median, 42.5 years; range, 20-64 years). The 43 control subjects were age- and sex matched, and had a mean age of 48.5 years (standard deviation, 10.9 years; median, 51.0 years; range, 16-63 years; P>0.05). The male-to-female ratio was confirmed as similar between the two groups (10:26 versus 12:31; P>0.05). All patients were clinically free from cardiovascular disease. Patients with a history of underlying valvular disease from any cause were excluded from the study. All patients underwent echocardiographic assessment, and the presence of any valvular lesions was documented. RESULTS: The mean duration of dexfenfluramine use by the cases was 21.8 weeks (standard deviation, 29.0 weeks; median, 18.0 weeks; range, 1-160 weeks). Subjects with a history of dexfenfluramine use had higher rates of significant aortic regurgitation, tricuspid regurgitation of any severity, and of any valvular regurgitation, compared to controls (11.1% versus 0%, P<0.05; 30.6% versus 4.7%, P<0.01; and 61.1% versus 34.9%, P<0.05, respectively). Logistic stepwise regression analysis to predict the risk of valvular lesion was conducted, with age, sex, history of dexfenfluramine use, body mass index, waist-hip ratio, blood pressure, fasting plasma glucose, lipid profile, and duration of diabetes as independent variables. A history of dexfenfluramine use was the only significant parameter entered into the model (significant aortic regurgitation: beta=9.19, standard error=46.6, P<0.05; any tricuspid regurgitation: beta=2.76, standard error=10.8, P<0.05). CONCLUSION: In Chinese patients with type 2 diabetes, a history of dexfenfluramine use is associated with heart-valve regurgitation, particularly aortic regurgitation. PMID- 12904611 TI - Patients' recommendation of doctor as an indicator of patient satisfaction. AB - OBJECTIVES: To determine whether patients' recommendation of their family doctor to others correlates with patient satisfaction scores, and to investigate other factors influencing patients' recommendation of doctor. DESIGN: Self-administered patient questionnaire. SETTING: Thirty-six family practice clinics, Slovenia. PATIENTS AND METHODS: A total of 2160 consecutive adult patients attending the clinics were approached to complete a self-administered questionnaire, to be returned in a prepaid envelope. The questionnaire included validated tools, such as the EUROPEP questionnaire on patient satisfaction and gathered data on health related quality of life, patient demographic, socio-economic and health characteristics, and attitudes and experience of health services. Patients also selected a response to the statement "I can strongly recommend my family doctor to my friends" on a five-point scale, from strongly disagree to strongly agree. RESULTS: Overall, 92% of respondents were in agreement with the statement that they would strongly recommend their family doctor to their friends. Patient satisfaction in the group of patients strongly agreeing with the statement was 11.1 points higher than that for the group responding they agreed only (92.4 versus 81.3 points; P<0.001). Multivariate analysis showed that 51.5% of the variation in the response to the statement could be explained by patient, doctor, and practice characteristics investigated. CONCLUSION: Higher agreement with the statement "I can strongly recommend my family doctor to my friends" was shown to be associated with higher patient satisfaction with the doctor's working style, with some patient demographic characteristics, aspects of patients' health care utilisation, and some doctor characteristics. Using a simple question regarding recommendation of the doctor to friends can be used as a surrogate measure of patient satisfaction, but should be interpreted with caution. PMID- 12904609 TI - Needlescopic cholecystectomy: prospective study of 150 patients. AB - OBJECTIVE: To evaluate the feasibility and safety of cholecystectomy using miniaturised instruments of 3 mm or less in diameter. DESIGN: Prospective study on patients with gallstones, with or without related complications. SETTING: Private hospital, Hong Kong. PATIENTS: From September 1997 to September 2002, 150 of the 180 consecutive patients managed were included in the present study. MAIN OUTCOME MEASURES: All patients were operated on with a standard four-port technique. Mini-laparoscopes of different sizes were used throughout the study period, which included 2-mm (n=33) fibre-optic laparoscope, and 2.5-mm (n=61) and 3-mm (n=56) laparoscopes of Hopkins rod lens system. The cystic duct and artery were secured either by extracorporeal ties or 10-mm clips passed through the umbilicus. The time taken from dissection to division of the cystic duct and artery, and to complete the operation were documented. RESULTS: The operation was successfully completed with needlescopic instruments in 127 (85%) patients, even though patients with acute cholecystitis and history of common bile duct stones were included. Use of larger-diameter mini-laparoscopes decreased the time needed to divide the cystic duct and artery, to detach the gall bladder from the liver, and to complete the operation. There were no deaths. One minor bile duct injury developed secondary to extensive cauterisation of the gall bladder fossa. CONCLUSION: Needlescopic cholecystectomy with minor technical modification can be completed within a duration comparable to standard laparoscopy at no increased risk for the great majority of patients with gallstones. PMID- 12904612 TI - A prevalence study of epilepsy in Hong Kong. AB - OBJECTIVES: To examine epidemiological data on epilepsy for the Hong Kong west region. DESIGN. Descriptive study. SETTING: Epilepsy clinic, university teaching hospital, Hong Kong. PATIENTS AND METHODS: The epilepsy clinic of Queen Mary Hospital manages the majority of adult patients (aged 15 years or older) with chronic seizure disorders resident in the Hong Kong west area with an adult population of 475,900. All patients underwent electroencephalography examination and each subject was independently assessed by two epileptologists for diagnosis and classified according to the International League Against Epilepsy recommendations. RESULTS: Seven hundred and thirty-six patients (female, 42.9%; male, 57.1%; mean age, 40.8 years; standard deviation, 13.6 years) with epilepsy were enrolled in the study. The prevalence rate of active epilepsy in the population 15 years or older was estimated at 1.54 per 1000 on 1 January 2002. Two hundred and eighty-five (38.7%) patients had idiopathic epilepsy syndromes, 100 (13.6%) had cryptogenic epilepsy, and 285 (38.7%) had a remote symptomatic aetiology. Seizure type was partial in 408 (55.4%) patients and generalised in 285 (38.7%). Thirty-one (4.2%) patients had a positive family history. Idiopathic generalised epilepsy syndromes described as common in the literature, such as juvenile myoclonic epilepsy and childhood absence epilepsy, were infrequently seen at 0.68% and 0.95% of cases, respectively. CONCLUSIONS: This study provides baseline data for epilepsy service development and research in Hong Kong. The prevalence rate of active epilepsy in this Chinese, adult population was low compared with that reported in other developed countries. Further population based epidemiological research is indicated to confirm the prevalence of seizure disorders in this locality. PMID- 12904613 TI - Symptomatic venous thromboembolism in Hong Kong Chinese children. AB - OBJECTIVE: To determine the incidence of venous thromboembolic disease in children of Chinese origin, and associated predisposing factors. DESIGN. Retrospective case series. SETTING: A general, public hospital serving a population of approximately 181,000 children in Hong Kong. PATIENTS AND METHODS: Hong Kong Chinese children under the age of 15 years who were diagnosed with a symptomatic venous thromboembolic event between 1995 and 2000 were included. Data on clinical features, predisposing factors, treatment, and outcome were obtained from review of hospital medical records. RESULTS: Eight children (five girls and three boys) of mean age 11.5 years (range, 0-14.7 years) were included in the study. They presented with deep vein thrombosis (n=4, with pulmonary embolism in one), superior vena cava thrombosis (n=1), and cerebral venous sinus thrombosis (n=3). Predisposing factors included hereditary protein C deficiency (n=3), protein S deficiency (n=2), anticardiolipin antibodies (n=1), malignancy (n=3), recent neurosurgery (n=2), infection (n=1), with multiple predisposing factors seen in three patients. Anticoagulant therapy was prescribed in five patients, and long-term warfarin therapy was required in two cases. Venous thromboembolic disease resolved in all children, but one patient had a recurrence after cessation of warfarin therapy, and one patient had post-thrombotic syndrome. CONCLUSION: The rate of venous thromboembolic disease in Hong Kong Chinese children was comparable to that seen in Caucasian children, with an annual incidence of 0.74 per 100,000 children. Predisposing factors, including hereditary prothrombotic conditions, were common. PMID- 12904614 TI - Prevalence and predictors of default from tuberculosis treatment in Hong Kong. AB - OBJECTIVE: To determine the prevalence and risk factors of default from tuberculosis treatment in Hong Kong. DESIGN: Retrospective study. SETTING: Data were obtained from programme forms completed by physicians in the Hong Kong Government Tuberculosis and Chest Service and from medical records from Hong Kong chest clinics. PATIENTS: In all, 5917 patients registered for antituberculous drug therapy in 1996; medical records of 5757 patients were reviewed. MAIN OUTCOME MEASURES: Patients who defaulted treatment were defined as those who had failed to collect medication for more than 2 consecutive months after the date of the last attendance during the course of treatment. Demographic and clinical characteristics, including history, treatment, and outcome, were compared between defaulters and non-defaulters, both among the whole group and among those with pulmonary disease. RESULTS: There were 442 (8%) patients who defaulted from treatment. Forty-five percent of those who defaulted did so in the first 2 months of treatment. Key risk factors associated with non-compliance were a history of default, male sex, and a history of concomitant liver disease or lung cancer. Among patients with pulmonary tuberculosis (381 defaulters and 1537 non defaulters), multiple drug resistance was also associated with default from treatment. Among defaulters with pulmonary disease, 39% were still bacteriologically positive at the time of default. CONCLUSION: Default from treatment may be partially responsible for the persistent high rates of tuberculosis in Hong Kong in the past decade. Health professionals should ensure that all barriers to treatment be removed and that incentives be used to encourage treatment compliance. PMID- 12904615 TI - The effects of tobacco use on oral health. AB - OBJECTIVE: To review the effects of tobacco use on oral health, with particular emphasis on the effects of periodontal diseases, dental implant failures, and risk of development of oral cancers and precancers. DATA SOURCES: Medline literature search (1977-2002). STUDY SELECTION: Key words for the literature search were 'tobacco smoking', 'periodontal disease', 'dental implant', and 'oral cancers and precancers'. DATA EXTRACTION: Evidence-based literature review. DATA SYNTHESIS: The prevalence and severity of periodontal diseases in their various forms are higher among smokers than among non-smokers. The success of dental implant treatments is significantly influenced by addiction to tobacco smoking. The failure rate of implant osseointegration is considerably higher among smokers, and maintenance of oral hygiene around the implants and risk of peri implantitis are adversely affected by smoking. The risks of developing oral cancers and precancers are greater in smokers. Betel nut chewing and smokeless tobacco produce similar risk to cancer incidence as tobacco smoking. Cessation of tobacco use has a beneficial effect on halting the progression of periodontal diseases and on the outcome of periodontal therapy. CONCLUSIONS: Medical and dental teams should be aware of oral problems associated with tobacco use. Counselling on smoking cessation and smoking prevention programmes should be an integral component of medical and dental teaching and practice. PMID- 12904616 TI - Subfascial endoscopic perforator surgery for venous ulcers. AB - We report the treatment and outcomes of 12 patients who underwent subfascial endoscopic perforator surgery for severe chronic venous insufficiency and venous ulceration. All patients had received prior superficial venous ablative surgery and presented with incompetent perforating veins in the calf and persistent venous ulceration (lasting >10 years). Outcome measures included ulcer healing time, recurrence, clinical symptom, and disability scores. There was one wound complication after subfascial endoscopic perforator surgery. The cumulative ulcer healing rate was 25% at 3 months, 42% at 6 months, and 92% at 1 year. One patient developed ulcer recurrence at 12 months after surgery. The mean clinical score and disability score decreased from 13.00 (standard deviation, 2.26) to 4.83 (1.47) and 1.75 (0.45) to 0.50 (0.52), respectively (P<0.001) after a median follow-up of 15.0 months (interquartile range, 12.0-21.5 months). Subfascial endoscopic perforator surgery was safe and effective in the treatment of patients with severe chronic venous insufficiency and venous ulceration caused by incompetent perforating veins in the calf. PMID- 12904617 TI - A practical guide to capacity assessment and patient consent in Hong Kong. AB - A person's decision-making capacity is pivotal when a doctor determines whether a person has the ability to make his or her own decisions, and thus it has major clinical, ethical, and legal significance, especially in Hong Kong, where the Mental Health Ordinance has introduced Guardianship provisions and provisions for the treatment of mentally incapacitated persons. This paper examines the legal concept of decision-making capacity and its clinical assessment within the local context. It is important for doctors to be aware of the legal mechanisms under which both urgent and non-urgent treatment can be given to mentally incapacitated persons, provided that the treatment is necessary and given in the best interests of the patient. PMID- 12904618 TI - Subtle perforation of the oesophagus by a foreign body. AB - A case of subtle oesophageal perforation caused by a foreign body is described. A 48-year-old woman had had a chicken bone impacted in the upper oesophagus for 4 days. At presentation, the bone was dislodged at endoscopy, and two small round depressions at opposite sides of the oesophageal wall were visible. The chest X ray findings were normal. Computed tomography of the thorax detected a small amount of air in the mediastinum. The water-soluble contrast swallow test showed no evidence of leakage. The patient was successfully treated using conservative measures. PMID- 12904619 TI - Use of botulinum toxin type A in a case of persistent parotid sialocele. AB - Sialocele is an uncommon complication of parotidectomy. Most cases resolve after conservative therapy consisting of repeated aspiration and pressure dressing. The condition is, however, occasionally resistant to such therapy. We report on a 52 year-old Chinese man who had a 10-year history of right parotid swelling. Following fine-needle aspiration cytology, Warthin's tumour was diagnosed, but after elective parotidectomy, a swelling developed and parotid sialocele was diagnosed. Botulinum toxin type A was given after the sialocele had persisted for almost 3 weeks after surgery, and after conservative management had been tried; the sialocele disappeared after two doses of treatment. Botulinum toxin therapy was thus an effective method of treating persistent sialocele. PMID- 12904620 TI - Cholangiocarcinoma presenting as pseudoachalasia and gastroparesis. AB - Cholangiocarcinoma most commonly presents as painless progressive jaundice. We report a case occurring in a 56-year-old Chinese woman with an unusual presentation of progressive dysphagia and vomiting. Oesophageal manometric and barium studies were indicative of achalasia, and computed tomography confirmed the presence of cholangiocarcinoma extending to the gastroesophageal junction and proximal lesser curve of the stomach. In this case, a constricting tumour at the gastroesophageal junction with probable invasion of the vagus nerves led to features of achalasia and gastroparesis. PMID- 12904621 TI - Electronic clinical practice guidelines: current status and future prospects in Hong Kong. PMID- 12904622 TI - The SARS attack on Hong Kong. PMID- 12904623 TI - Severe acute respiratory syndrome and biology, air quality, physics, and mechanical engineering. PMID- 12904624 TI - War on Iraq: the public health perspective. PMID- 12904625 TI - Propaganda or science? Biological warfare and the people of Iraq. PMID- 12904626 TI - Treatment of severe acute respiratory syndrome with convalescent plasma. PMID- 12904627 TI - Treating emerging viral infections. PMID- 12904629 TI - Modelling the severe acute respiratory syndrome epidemic. PMID- 12904630 TI - Race and ethnicity in medical research. PMID- 12904631 TI - Prediction of 200-m sprint kayaking performance. AB - The aim of this study was to determine the anthropometric and physiological profile of 200-m sprint kayakers and to examine relationships with 200-m race performance. Twenty-six male kayakers who were categorised in two ability groups, international (Int) and national (Nat) level, underwent a battery of anthropometric and physiological tests and a 200-m race. Race time was significantly lower in Int than Nat (39.9 +/- 0.8 s and 42.6 +/- 0.9 s, respectively). Int demonstrated significantly greater measures of mesomorphy, biepycondylar humeral breadth, circumferences of the upper arm, forearm and chest, peak power and total work in a modified Wingate test, total work in a 2 min ergometry test, peak isokinetic power, and peak isometric force. Significant relationships were found between 200-m time and a number of anthropometric variables and anaerobic and dynamometric parameters. Stepwise multiple regression revealed that total work in the modified Wingate alone predicted 200-m race time (R2 = 0.53, SEE = 1.11 s) for all 26 subjects, while biepycondylar humeral breadth alone predicted race time (R2 = 0.54, SEE = 0.52 s) in Int. These results demonstrate that superior upper body dimensions and anaerobic capacities distinguish international-level kayakers from national-level athletes and may be used to predict 200-m performance. PMID- 12904632 TI - Development of race profiles for the performance of a simulated 2000-m rowing race. AB - The purpose of this study was to determine the race profile for a 2000-m simulated rowing race as well as the effect of training and gender on the race profile. Nineteen men and 19 women undertook a 2000-m simulated rowing race before and after 10 weeks of a typical off-season training program for rowing. Velocity was calculated every 200 m and the deviation in velocities from the mean race velocity (MRV) was plotted every 200 m to produce race profiles for each gender before and after training. The three fastest male rowers varied approximately 0.02 m.s from the MRV after training and displayed a constant-pace model. The fastest female rowers displayed an all-out strategy after training, producing large deviations from MRV. Average squared deviations from the mean (SDM) determined that all groups except the fastest females had a reduction in MRV deviation after training. These results suggest that the optimal race profile for a simulated 2000-m rowing race may be different between genders. Training reduces SDM and influences both male and female pacing patterns such that both exhibit a pacing strategy that is more similar to that of elite athletes in other events of similar and shorter duration. PMID- 12904633 TI - Prediction of drafted-triathlon race time from submaximal laboratory testing in elite triathletes. AB - PURPOSE AND METHODS: To determine which physiological variables accurately predict the race time of an Olympic-distance International Triathlon undertaken in drafted conditions, 8 elite triathletes underwent both maximal and submaximal laboratory and field physiological testing: a 400-m maximal swim test; an incremental treadmill test; an incremental cycling test; 30 min of cycling followed by 20 min of running (C-R); and 20 min of control running (R) at the exact same speed variations as in running in C-R. Blood samples were drawn to measure venous lactate concentration after the 400-m swim and the cycle and run segments of C-R. During the maximal cycling and running exercises, data were collected using an automated breath-by-breath system. RESULTS: The only parameters correlated with the overall drafted-triathlon time were lactate concentration noted at the end of the cycle segment (r = 0.83, p < 0. 05) and the distance covered during the running part of the submaximal C-R test (r = 0.92, p < 0. 01). Stepwise multiple regression analysis revealed a highly significant (r = 0.96, p < 0.02) relationship between predicted race time (from laboratory measures) and actual race time, using the following calculation: Predicted Triathlon Time (s) = 1.128 (distance covered during R of C-R [m]) + 38.8 ([lactate] at the end of C in C-R) + 13,338. The high R2 value of 0.93 indicated that, taken together, these two laboratory measures could account for 93% of the variance in race times during a drafted triathlon. CONCLUSION: Complementing previous studies, this study demonstrates that different parameters seem to be reliable for predicting performance in drafted vs. nondrafted Olympic-triathlon races. It also demonstrates that, for elite triathletes competing in a drafted Olympic-distance triathlon, performance is accurately predicted from the results of submaximal laboratory measures. PMID- 12904634 TI - Critical velocity and anaerobic distance capacity in prepubertal children. AB - This study was designed to calculate the critical velocity (vcrit) and anaerobic distance capacity (ADC) of prepubescent children for running events. Thirty-four prepubertal children underwent a graded field test to exhaustion in order to determine peak oxygen uptake (peak VO2) and maximal aerobic velocity (MAV). Then, in random order, they performed five runs to exhaustion (tlim) at relative velocities corresponding to 90, 95, 100, 105, and 110% of MAV. The linear relationships between distance limit (dlim) and tlim were calculated in order to determine vcrit (slope of the relationship) and ADC (intercept). Very high individual coefficients of determination were found between dlim and tlim (0.98 < r2 < 0.99; p < 0.001). The vcrit was significantly correlated with peak VO2 (r = 0.73; p < 0.001). However, no relationship was found between ADC and the maximal accumulated oxygen deficit. In conclusion, our results indicated that, for children, the relationship between dlim and tlim could be calculated with tlim ranging from 2 to 10 min, and that vcrit is a good indicator of the aerobic fitness of children. Nevertheless, further studies will have to be conducted to validate the use of ADC as an indicator of children's anaerobic capacity. PMID- 12904635 TI - Effects of different stepwise reduction taper protocols on cycling performance. AB - This study examined the effects of different 7-day taper protocols on simulated 20-km time trials (20TT). Following 3 weeks of baseline training, 11 male cyclists (.VO2max = 4.78 +/- 0.66 L.min-1) were randomly assigned to one of three stepwise reduction tapers in which training volume was reduced by 30% (T30, n = 5), 50% (T50, n = 6), or 80% (T80, n = 6) of baseline training with intensity (85% .VO2max) maintained. Cardiorespiratory measurements were collected every 5 km during the 20TT. Results revealed a significant (5.4%, 0.05) improvement in 20TT performance in the T50 protocol with concomitant increases in .VO2 and O2 pulse. No significant differences were found in T30 or T80. These results showed that a moderate (50%) reduction in weekly training volume appeared to be optimal in terms of enhancing performance. This confirms the contention that proper placement of training volume during tapering, while maintaining exercise intensity, can elicit performance improvements. PMID- 12904636 TI - Overloaded training increases exercise-induced oxidative stress and damage. AB - We hypothesized that overloaded training (OT) in triathlon would induce oxidative stress and damage on muscle and DNA. Nine male triathletes and 6 male sedentary subjects participated in this study. Before and after a 4-week OT, triathletes exercised for a duathlon. Blood ratio of reduced vs. oxidized glutathione (GSH/GSSG), plasma thiobarbituric acid reactive substances (TBARS), leukocyte DNA damage, creatine kinase (CK), and CK-MB mass in plasma, erythrocyte superoxide dismutase (SOD) activity, erythrocyte and plasma glutathione peroxidase (GSH-Px) activities, and plasma total antioxidant status (TAS) were measured before and after OT in pre- and postexercise situations. Triathletes were overloaded in response to OT. In rest conditions, OT induced plasma GSH-Px activity increase and plasma TAS decrease (both p < 0.05). In exercise conditions, OT resulted in higher exercise-induced variations of blood GSH/GSSG ratio, TBARS level (both p < 0.05), and CK-MB mass (p < 0.01) in plasma; and decreased TAS response (p < 0.05). OT could compromise the antioxidant defense mechanism with respect to exercise-induced response. The resulting increased exercise-induced oxidative stress and further cellular susceptibility to damage needs more study. PMID- 12904637 TI - Reproducibility of the cycling time to exhaustion at .VO2peak in highly trained cyclists. AB - The purpose of the present study was to examine, in highly trained cyclists, the reproducibility of cycling time to exhaustion (T(max)) at the power output equal to that attained at peak oxygen uptake (.VO2peak) during a progressive exercise test. Forty-three highly trained male cyclists (M +/- SD; age = 25 +/- 6 yrs; weight = 75 +/- 7 kg; .VO2peak = 64.8 +/- 5.2 ml.kg-1.min-1) performed two T(max) tests one week apart. While the two measures of T(max) were strongly related (r = 0.884; p < 0.001), T(max) from the second test (245 +/- 57 s) was significantly higher than that of the first (237 +/- 57 s; p = 0.047; two-tailed). Within subject variability in the present study was calculated to be 6 +/- 6%, which was lower than that previously reported for T(max) in sub-elite runners (25%). The mean T(max) was significantly (p < 0.05) related to both the second ventilatory turnpoint (VT(2); r = 0.38) and to .VO2peak (r = 0.34). Despite a relatively low within-subject coefficient of variation, these data demonstrate that the second score in a series of two T(max) tests may be significantly greater than the first. Moreover, the present data show that T(max) in highly trained cyclists is moderately related to VT(2) and .VO2peak. PMID- 12904638 TI - Prolonged daytime exercise repeated over 4 days increases sleeping heart rate and metabolic rate. AB - The aim of this study was to determine the influence of prolonged exercise repeated for 4 days on sleeping heart rate (SHR) and metabolic rate (SMR). Eleven young untrained men exercised at moderate intensity 5 hrs daily for 4 days, alternately on a cycle ergometer (57.0 +/- 1.3% .VO2max) and a treadmill (64.7 +/ 1.6% .VO2max). They spent the night prior to the exercise period (control, C) and the 4 nights following exercise days (N1 to N4) in room calorimeters for the measurement of SHR, SMR, and respiratory quotient (RQ) from midnight until 6 a.m. Every morning, before the exercise bouts, plasma-free epinephrine (E) and norepinephrine (NE) levels were measured. After exercise, all SHR values were significantly higher than at C level (52 +/- 1 bpm, p < 0.001) and the highest value was observed on N2 (61 +/- 2 bpm). SMR increased by 11.2 +/- 1.5% from C to N1, p < 0.001, and then plateaued up to N4, whereas RQ decreased from C (0.833 +/ 0.009) to N2 (0.798 +/- 0.005) and then plateaued. Plasma NE levels were higher the morning after each day of exercise and peaked on N2, whereas no significant variations were found for E. Variations of SHR between C and N2, and N3 and N4 were correlated with changes of SMR. No significant relationships were found between morning plasma NE, and either SMR or SHR variations. To conclude, prolonged exercise repeated for 4 days was associated with increases in SHR and SMR during the night following each day of exercise concomitantly with an enhanced lipid oxidation. The sustained stimulation of the sympathetic nervous system may be partly responsible for these effects. PMID- 12904639 TI - Activity patterns during pregnancy. AB - The purpose of this study was to investigate determinants of the activity patterns of women prior to pregnancy and factors associated with quitting activities during pregnancy. METHODS: These data arose from a study designed to look at the impact of exercise in pregnancy on birth weight (Campbell and Mottola, 2001). This secondary analysis explored relationships between subject characteristics and exercise patterns via a self-completed questionnaire. Univariable and multivariable odds ratios were estimated using logistic regression. Multivariable models used backward stepwise variable selection. RESULTS: A total of 853 women agreed to participate and 529 women (62%) returned completed questionnaires. Of these, 369 (70%) and 258 (49%) engaged in a structured exercise program before pregnancy and in Trimester 3, respectively. Factors associated with engaging in regular structured exercise prior to pregnancy included: postsecondary education (OR = 1.50; 0.98, 2.30), no children (OR = 2.44; 1.56, 3.82), nonsmoker (OR = 1.84; 1.18, 2.88), and involvement in regular recreational activities (OR = 3.07; 1.81, 5.20). During pregnancy, all categories of activity decreased except walking, which increased by Trimester 3. Factors associated with quitting a regular structured exercise program by Trimester 3 were: having children (OR = 1.67; 1.05, 2.67), a prepregnancy BMI of 25 (OR = 1.79; 1.04, 3.13), and higher weight gain. IMPLICATIONS: Community programs that encourage active living should address these factors. PMID- 12904640 TI - Training profile counts for time-to-exhaustion performance. AB - The objective of this study was to compare the time to exhaustion (Tlim) at maximal aerobic speed (v.VO2max) in middle- and long-distance runners. Five middle-distance (MDR) and 5 long-distance (LDR) male runners, ages 28 +/- 7 years, were tested running on a treadmill, with the Universite de Montreal Track Test (UMTT), on maximal velocity and on time-to-exhaustion track tests. During the laboratory test, cardiorespiratory variables (e.g., HR, .VO2max, .VCO2, .VE) were assessed. Second, running velocity at .VO2max (v.VO2max) during the UMTT was determined and HR values were recorded; also, maximal velocity on a 30-m sprint (V30) and maximal heart rate (HR max) and time to exhaustion were determined on the track. No significant difference was observed between groups during the multistage treadmill test. Significant differences were found between groups for V30 and Tlim, with MDR showing a 23% longer running time than LDR. The results of the present study demonstrated that the training profile of middle-distance and long-distance runners plays a significant role in Tlim performance when v.VO2max is obtained during a test with short-duration stages. PMID- 12904641 TI - The function and organization of plasmids. PMID- 12904642 TI - Choosing a cloning vector. PMID- 12904644 TI - Chemical transformation of E. coli. PMID- 12904643 TI - Escherichia coli host strains. PMID- 12904645 TI - Electroporation of E. coli. PMID- 12904646 TI - DNA transfer by bacterial conjugation. PMID- 12904647 TI - Cosmid packaging and infection of E. coli. PMID- 12904648 TI - Isolation of plasmids from E. coli by alkaline lysis. PMID- 12904649 TI - Isolation of plasmids from E. coli by boiling lysis. PMID- 12904650 TI - High-purity plasmid isolation using silica oxide. PMID- 12904651 TI - High-throughput plasmid extraction using microtiter plates. PMID- 12904652 TI - Isolation of cosmid and BAC DNA from E. coli. PMID- 12904653 TI - Preparation of single-stranded DNA from phagemid vectors. PMID- 12904654 TI - Using desktop cloning software to plan, track, and evaluate cloning projects. PMID- 12904655 TI - Cloning in plasmid vectors. PMID- 12904656 TI - Extraction of DNA from agarose gels. PMID- 12904658 TI - Construction of genomic libraries in lambda-vectors. PMID- 12904659 TI - Rapid screening of recombinant plasmids. PMID- 12904657 TI - Cloning PCR products with T-vectors. PMID- 12904660 TI - Restriction analysis of recombinant plasmids. PMID- 12904661 TI - Screening recombinant DNA libraries. PMID- 12904662 TI - Sequencing using fluorescent-labeled nucleotides. PMID- 12904663 TI - Site-directed mutagenesis using the megaprimer method. PMID- 12904664 TI - Site-directed mutagenesis by inverse PCR. PMID- 12904665 TI - Creating nested DNA deletions using exonuclease III. PMID- 12904666 TI - Transposon and transposome mutagenesis of plasmids, cosmids, and BACs. PMID- 12904667 TI - In vitro transcription and translation. PMID- 12904668 TI - Vectors for the expression of recombinant proteins in E. coli. PMID- 12904669 TI - Expression of recombinant proteins from lac promoters. PMID- 12904670 TI - Plasmid-based reporter genes: assays for beta-galactosidase and alkaline phosphatase activities. PMID- 12904671 TI - Plasmid-based reporter genes: assays for green fluorescent protein. PMID- 12904672 TI - Conditional pairwise estimation in the Rasch model for ordered response categories using principal components. AB - In the Rasch model for items with more than two ordered response categories, the thresholds that define the successive categories are an integral part of the structure of each item in that the probability of the response in any category is a function of all thresholds, not just the thresholds between any two categories. This paper describes a method of estimation for the Rasch model that takes advantage of this structure. In particular, instead of estimating the thresholds directly, it estimates the principal components of the thresholds, from which threshold estimates are then recovered. The principal components are estimated using a pairwise maximum likelihood algorithm which specialises to the well known algorithm for dichotomous items. The method of estimation has three advantageous properties. First, by considering items in all possible pairs, sufficiency in the Rasch model is exploited with the person parameter conditioned out in estimating the item parameters, and by analogy to the pairwise algorithm for dichotomous items, the estimates appear to be consistent, though unlike for the dichotomous case, no formal proof has yet been provided. Second, the estimates of each item parameter is a function of frequencies in all categories of the item rather than just a function of frequencies of two adjacent categories. This stabilizes estimates in the presence of low frequency data. Third, the procedure accounts readily for missing data. All of these properties are important when the model is used for constructing variables from large scale data sets which must account for structurally missing data. A simulation study shows that the quality of the estimates is excellent. PMID- 12904673 TI - Reliability and true-score measures of binary items as a function of their Rasch difficulty parameter. AB - This article provides formulas for expected true-score measures and reliability of binary items as a function of their Rasch difficulty when the trait (ability) distribution is normal or logistic. The proposed formulas have theoretical value and can be useful in test development, score analysis, and simulation studies. Once the items are calibrated with the dichotomous Rasch model, one can estimate (without further data collection) the expected values for true-score measures (e.g., domain score, true score variance, and error variance for the number-right score) and reliability for both norm-referenced and criterion-referenced interpretations. Thus, given a bank of Rasch calibrated items, one can develop a test with desirable values of population true-score measures and reliability or compare such measures for subsets of items that are grouped by substantive characteristics (e.g., content areas or strands of learning outcomes). An illustrative example for using the proposed formulas is also provided. PMID- 12904674 TI - Using logistic regression to detect item-level non-response bias in surveys. AB - This article describes a procedure for evaluating item-level non-response bias in questionnaire items. Specifically, logistic regression is used to determine whether non-responses are random or systematic in nature for one question from the National Educational Longitudinal Study of 1994 concerning drug use behaviors. It turns out that, indeed, non-responses are systematic with males and lower achieving students being more likely to contribute to non-response along with two-way interactions between ethnicity and SES and ethnicity and geographic region. In addition, the magnitude of the potential bias is estimated, which demonstrates that the parameter estimates obtained by assuming that the data are missing at random may be extremely biased, given this frame of reference. Finally, several steps are suggested for evaluating the threat of non-response bias in survey research. PMID- 12904675 TI - Rasch measurement in the assessment of amytrophic lateral sclerosis patients. AB - This paper examines the sensitivity to change over time of the Amyotrophic Lateral Sclerosis Assessment Questionnaire (ALSAQ-40). Individuals' health status change was assessed by means of the Rasch-based Reliable Change Index (RCI) for ALSAQ-40 questionnaires completed on two occasions, three months apart. In addition, at follow-up respondents indicated how much change they had experienced since baseline via dimension-specific self reported transition questions. 764 individuals returned questionnaires at baseline and follow-up. For all dimensions, of respondents defined by the RCI as worse, a majority rated themselves as worse. However, on two dimensions over 60% of the respondents who rated themselves as being worse were defined as unchanged by the RCI. As with effect size smaller RCI cut-off points might be needed for subjects with ALS. This study confirms that the ALSAQ-40 is a valid and responsive disease specific health related quality of life instrument for use in studies of patients with ALS or other motor neuron diseases. PMID- 12904677 TI - The recovery of the density scale using a stochastic quasi-realization of additive conjoint measurement. AB - This paper attempts to illuminate some of the practical limitations that the Rasch model (and by extension, Item Response Theory models) may have by focusing on the recovery of the density scale. Five simulation trials were conducted: the first four to recover the density scale with different deviations from the assumptions implicit in the use of the Rasch model and the fifth trial with an almost ideal data set. Results demonstrate that when error distributions are insufficient the results may be ordinal at best, and when error distributions are non-symmetrical, the positions of items may be biased with respect to the positions of persons. Results also confirm that errors of estimation, and test and sample information functions are sample dependent. PMID- 12904676 TI - Measuring client satisfaction with public education II: comparing schools with state benchmarks. AB - Because the results of the client satisfaction evaluation trials conducted in the state's public schools revealed that levels of client satisfaction differed in significant and meaningful ways between parents and students as well as between types of schools, this research consultancy provided school versus benchmark comparison reports based on groups of generally comparable school types. The state benchmark for each set of comparable schools estimated how easy it was, on average, for the members of the benchmark group (parents or students) to endorse each of the 20 School Opinion Survey Likert-scale items (King and Bond, 2003). The school satisfaction level for each item was calculated by a similar process to estimate how easy it was, on average, for the members of the school sample (parents or students) to endorse that item. Reports to individual schools used easy to interpret Parent/Student Satisfaction Graphs which plotted the Rasch modeled differences between the state benchmark level and the school level for each item. In very small schools where the small amount of data did not allow for item-by-item graphs to be constructed, overall satisfaction graphs provided one global comparison with the appropriate benchmark to be reported. PMID- 12904678 TI - Substantive scale construction. AB - Variable construction requires careful attention to substantive issues; a theory guiding its development, a hierarchy of illustrative items constructed to define the variable, the subsequent production of item difficulties and person measures, and the analysis of fit. Rasch measurement practitioners should give careful attention to these matters so practical suggestions are given for designing variables based on theory, item construction, and Rasch models for the analysis of data. Variable maps are emphasized to guide variable construction and interpret the results. PMID- 12904680 TI - Crossed and uncrossed olivocochlear pathways exacerbate temporary shifts in hearing sensitivity after narrow band sound trauma in normal ears of animals with unilateral hearing impairment. AB - Olivocochlear (OC) pathways have been shown to reduce the temporary threshold shifts (TTSs) caused by traumatic sounds. More recently they have been shown to exacerbate TTSs under certain conditions. One condition is the normal-hearing ear of animals with a chronic unilateral hearing loss. Testing with pure tone trauma showed that then (a) the normal-hearing ear had a lower-than-normal 'intrinsic' susceptibility to intense tones, (b) binaural trauma exacerbated TTSs in the normal-hearing ear through the activity of uncrossed OC (UOC) pathways, and (c) there was no effect on TTSs of the crossed OC (COC) pathway to the normal-hearing ear. The present study is an examination in such animals of effects with noise band trauma. The effects here confirm the previous finding that under such conditions the normal-hearing ear has a lower-than-normal susceptibility to loud sound, and binaural loud sounds exacerbate TTSs in the normal-hearing ear. They extend the previous study by demonstrating that with this traumatic sound, both COC and UOC pathways exacerbate TTSs. These effects contrast against the effects seen in animals with bilaterally normal hearing for the same noise band. Given the commonality of unilateral hearing losses in the normal human population, these data have implications for the functional effects of the OC pathways on loud sound-induced hearing damage. PMID- 12904679 TI - Fas ligand expression in the organ of Corti. AB - We have previously demonstrated by FACS analysis and histochemistry that Fas ligand (FasL) increases on cochlear cell surfaces after immune response or stimulation with gamma-interferon (IFN-gamma). To determine whether the appearance of FasL on cochlear cell membranes is related to gene expression or to posttranslational events, cochlear cells were treated with IFN-gamma. They were evaluated for FasL gene expression by real-time PCR and for FasL protein localization by confocal microscopy of permeabilized and immunolabeled cells. Real-time PCR analysis of cDNAs generated from unstimulated or IFN-gamma stimulated organ of Corti demonstrated no change in the transcription of the gene encoding FasL. In contrast, confocal microscopy revealed dramatic changes in the cellular distribution of FasL, consistent with movement from the endoplasmic reticulum to the cytoplasm and cell membrane. The results suggest that recruitment of preformed FasL from intracellular compartments, rather than its biosynthesis, is responsible for the increase in FasL on the cell surface following IFN-gamma stimulation. This is similar to the response of cytotoxic T lymphocytes in which gene expression is not involved in FasL surface appearance. Presumably, the use of preformed FasL increases the rapidity of this response. FasL localization to the membrane may be involved in protecting the inner ear from autoimmunity or inflammation. Alternatively it may be related to cochlear cell death in response to inflammatory stress. PMID- 12904681 TI - Language and speech perception outcomes in hearing-impaired children with and without connexin 26 mutations. AB - This study addressed the question of whether a hearing loss caused by mutations in the connexin 26 gene had a significant effect on language and speech perception outcomes in children using cochlear implants or hearing aids. The families of children who had participated in two previous studies of language development were invited to participate in this genetic study. From the 52 children whose families agreed to participate, 15 children with connexin 26 mutations in both chromosomes were identified. After taking into account other factors which are known to affect language development and speech perception in children with impaired hearing, no significant differences were found between the 15 children where connexin 26 was known to be the cause of deafness and the other 37 children in the study. PMID- 12904682 TI - Acoustic and electrical pattern analysis of consonant perceptual cues used by cochlear implant users. AB - It is hypothesized that for postlingually deafened adult cochlear implant (CI) users, a significant source of their perceptual performance variability is attributable to differences in their ability to discriminate the basic perceptual cues that are important in speech recognition. Previous research on 'electric hearing' has identified consistent perceptual cues for vowel recognition. However, the results on consonant perception by CI users are less clear. The primary purpose of this study is to present a quantitative method of evaluating potential 'electric cues' used by CI users in consonant identification. Since the actual input signals to the auditory periphery of CI users are electric in nature, we elected to measure the CI electric discharge patterns in addition to the original acoustic waveforms. The characteristics of the electric discharge patterns in response to intervocalic consonants were quantified and correlated with the dimensions of CI patients' perceptual spaces, which were computed from multidimensional scaling analyses of their consonant confusion matrices. The results agree with most, but not all, commonly accepted acoustic cues used by normal-hearing listeners. The correlation findings also suggest that CI users employ different sets of 'electric cues' in perceiving consonants that differ in their manner of articulation. Specifically, spectral and temporal cues associated with slowly changing formant structures and transitions, and features associated with frication and high-frequency noise, are all highly correlated with the perceptual dimensions of all CI users. However, rapidly changing formant transitions, such as those present in stop consonants, did not appear to play a significant role in consonant recognition by more poorly performing CI subjects. The perceptual results were consistent with our physical findings that the SPEAK coding strategy partially degraded the rapidly changing formant transitions. PMID- 12904683 TI - Improvements in trunk sway observed for stance and gait tasks during recovery from an acute unilateral peripheral vestibular deficit. AB - OBJECTIVES: Our aim was to track improvements in postural control during recovery from an acute unilateral peripheral vestibular deficit (UVL), presumably due to vestibular neuritis, and to determine if recovery rates were different for stance and gait tasks. Postural control was quantified using simple measurements of trunk sway: amplitudes of trunk sway angle and angular velocity, in the roll and pitch directions as well as task duration, were examined for a battery of stance and gait tasks. These measures were collected at the onset of the deficit and then 3 weeks and 3 months later. STUDY DESIGN: A repeated-measures design was used for UVL subjects and age-matched healthy controls. Stance tasks involved standing on 1 or 2 legs with eyes open or closed. Gait tasks consisted of tandem gait, walking normally with eyes closed, or with the head rotating or head pitching, walking up and down stairs and walking over a series of low barriers. Stance and tandem gait tasks were repeated using a foam support surface instead of a normal floor. PATIENTS: Twenty-eight patients with acute UVL were examined. MAIN OUTCOME MEASURES: The range of trunk sway angular displacement and angular velocity in the pitch and roll directions was measured for each task in addition to task duration. The measures were compared with those of normal subjects. RESULTS: The amplitudes of pitch trunk sway for 2-legged stance tasks with eyes closed underwent the greatest reduction 3 weeks after UVL onset. At 3 months, trunk sway was almost normal for all 2-legged stance tasks. One-legged stance tasks with eyes open showed a similar but slower improvement. Stance time without a fall showed a very rapid improvement for 1-legged tasks but was still shorter than that of normal subjects at 3 months. Trunk sway for the simple gait tasks was within normal range at 3 months; however, task duration was still longer than normal. More complex gait tasks, such as walking 8 tandem steps on foam or walking up and down stairs, showed no improvement in trunk roll sway at 3 months. A mix of variables from mainly gait tasks best identified a balance deficit due to UVL, with complex gait tasks becoming more important for identification purposes as compensation progressed. The accuracy of UVL identification with durations alone was 75% of the accuracy with combined trunk sway and duration measures. CONCLUSIONS: These data suggest that recovery of normal trunk control during the compensation process for unilateral vestibular hypofunction is more rapid for stance tasks than gait tasks. Even at 3 months, trunk sway for complex gait tasks was not normal. Thus, trunk sway for gait tasks provides a better insight into remaining deficits in balance control of vestibular-loss patients than the sway of stance tasks. PMID- 12904684 TI - Effect of angiotensin II receptor blocker on plasma levels of TGF-beta 1 and interstitial fibrosis in hypertensive kidney transplant patients. AB - BACKGROUND/AIM: Transforming growth factor-beta1 (TGF-beta 1) is involved in the pathogenesis of chronic allograft nephropathy after kidney transplantation. The aim of the study was to evaluate the effect of the angiotensin receptor blocker losartan on TGF-beta 1 plasma levels and proteinuria in hypertensive transplant recipients. METHODS: A total of 162 transplant recipients were included in the study. The patients were randomized into 3 groups: group 1 received losartan; group II received an angiotensin-converting enzyme inhibitor (captopril), and group III received a calcium channel blocker (amlodipine). All the parameters were recorded at the time of therapy initiation and at 1, 4 and 12 weeks and 12 months thereafter. Graft biopsy before the start and at the end of the study was done to evaluate histopathological progression. RESULTS: Blood pressure was controlled in the 3 groups; however, the need for other antihypertensive agents was significant in groups I and II. Treatment with losartan significantly decreased the plasma level of TGF-beta1, 24-hour urinary protein and serum uric acid (p < 0.05). No significant changes were seen in the hemoglobin or serum potassium levels. The rate of histopathological progression was significantly lower in the losartan group. No patient was discharged from the study due to side effects. CONCLUSIONS: After transplantation all drugs were able to control blood pressure with good safety and tolerability. The study demonstrates that ARB significantly decreases the plasma levels of TGF-beta1, proteinuria and uric acid. These results could play an important and decisive role in the treatment and prevention of chronic allograft nephropathy. PMID- 12904685 TI - Ionotropic glutamate receptors in Caenorhabditis elegans. AB - Ionotropic glutamate receptors (iGluRs) are an important class of heteromeric ligand-gated receptor complexes that mediate a large portion of the excitatory neurotransmission in the vertebrate CNS. Since the cloning of the first iGluR subunit in 1989, the study of this receptor family has rapidly developed in mammals and expanded to include the study of conserved glutamate receptors in simpler invertebrate systems, including the fruit fly Drosophila melanogaster and the soil nematode Caenorhabditis elegans. These model organisms have enabled the genetic analysis of glutamate receptors in the context of a simpler nervous system and provided new insights into receptor function and regulation. In this review we will focus on recent studies that have used genetic, behavioral, and electrophysiological techniques to study the function of iGluRs in C. elegans. PMID- 12904686 TI - Identified ion channels in the squid nervous system. AB - Our modern understanding of channels as discrete voltage-sensitive and ion selective entities comes largely from a series of classical studies using the squid giant axon. This system has also been critical for understanding how transporters and synaptic transmission operate. This review outlines attempts to assign molecular identities to the extensively studied physiological properties of this system. As it turns out, this is no simple task. Molecular candidates for voltage-gated Na(+), K(+), and Ca(2+) channels, as well as ion transporters have been isolated from the squid nervous system. Both physiological and molecular approaches have been used to equate these cloned gene products with their native counterparts. In the case of the delayed rectifier K(+) conductance, the most thoroughly studied example, two major issues further complicate the equation. First, the ability of K(+) channel monomers to form heteromultimers with unique properties must be considered. Second, squid K(+) channel mRNAs are extensively edited, a process that can generate a wide variety of channel proteins from a common gene. The giant axon system is beginning to play an important role in understanding the biological relevance of this latter process. PMID- 12904687 TI - The ascidian dihydropyridine-resistant calcium channel as the prototype of chordate L-type calcium channel. AB - This review describes recent findings on voltage-gated Ca channel (Cav channel) cloned from ascidians, the most primitive chordates. Ascidian L-type like Cav channel has several unusual features: (1). it is closely related to the prototype of chordate L-type Cav channels by sequence alignment; (2). it is resistant to dihydropyridine due to single amino acid change in the pore region, and (3). maternally provided RNA putatively encodes a truncated protein which has remarkable suppressive effect on Cav channel expression during development. Ascidian Cav channel will provide a useful molecular clue in the future to understand Ca(2+)-regulated cell differentiation and physiology with the background of recently defined ascidian genome and molecular biological tools. PMID- 12904688 TI - Cells derived from regenerated endothelium of the porcine coronary artery contain more oxidized forms of apolipoprotein-B-100 without a modification in the uptake of oxidized LDL. AB - Increased accumulation of lipoproteins and cholesterol within cells from regenerated endothelium may be responsible for their reported dysfunction. This study compared the presence and uptake of oxidized forms of low-density lipoprotein (LDL) in cells derived from native and regenerated endothelium. Four weeks after balloon denudation, primary cultures of native and regenerated endothelial cells were prepared from porcine coronary arteries. Regenerated endothelium stained more strongly using an antibody against oxidized lipoproteins. The increase in oxidized forms of apolipoprotein-B-100 exhibited by cells from regenerated endothelium was not due to an increase in extracellular induced oxidation of native LDL, measured as the production of thiobarbituric acid-reactive substances, being identical in both cell types. Intracellular cholesterol and cholesterol ester content were unchanged in regenerated cells. Using flow cytometry, accumulation of oxidized LDL was investigated further by quantifying the uptake of a mildly oxidized preparation of 1,1'-dioctadecyl 3,3,3',3-tetramethyl-indocarbocyanine perchlorate-labelled LDL. The parameters of uptake, EC(50) and E(max), were not different between cells from native and regenerated endothelium suggesting that the number of LOX-1 receptors was identical in the two cell types. Moreover, a negative correlation between the increased uptake of acetylated LDL and decreased cGMP production in response to bradykinin was observed in cells from regenerated endothelium. Thus, the increased incorporation of modified LDL and their intracellular oxidation could be responsible for the alteration in NO production. The presence of oxidized forms of LDL may be a marker of endothelium regeneration and could be involved in the endothelial dysfunction of pig coronary arteries 4 weeks after balloon denudation. PMID- 12904689 TI - Preoperative assessment of myometrial invasion and cervical involvement of endometrial cancer by transvaginal ultrasonography. AB - The aim of the study was to assess the depth of myometrial invasion and cervical involvement by endometrial cancer using preoperative 6.5-MHz, high-frequency transvaginal ultrasonography as compared with postoperative assessment using histopathological examination. The study included 47 patients with histologically proven cancers of the endometrium. All patients underwent transvaginal sonography before surgery. The depth of myometrial invasion was classified as none, inner half of the uterine wall, and outer half of the uterine wall. Cervical spread is recorded as positive or negative. Of 36 (76.6%) patients with proven myometrial invasion, 33 cases (91.66%) were revealed by sonography. Histologically proven cervical invasion that correlated with sonography was shown in 3 patients (75%). In 7 patients (14.9%) ultrasonography could not correctly predict the depth of myometrial invasion. The depth of invasion was underestimated in 4 (8.5%) cases and overestimated in 3 (6.4%) cases. Preoperative assessment of invasion of the uterine wall and cervical spread by transvaginal ultrasonography had an accuracy of 85 and 97.8% if correlated with the definitive histopathological examination. The role of transvaginal ultrasonography in preoperative assessment of the depth of myometrial invasion and cervical involvement in patients with endometrial cancer needs to be studied further before making reliable conclusions. PMID- 12904690 TI - Heparanase expression and angiogenesis in endometrial cancer. AB - Human heparanase has been shown to function in tumor progression, metastatic spread, and tumor angiogenesis. The aim of the present study was to assess heparanase expression in endometrial cancer in correlation with neovascularization and clinicopathological factors. Forty endometrial cancers were obtained from previously untreated patients (median age 55.5, range 33-78 years). The expression of heparanase mRNA was evaluated using a semiquantitative reverse transcriptase-polymerase chain reaction. Tumor angiogenesis was assessed using microvessel counting. The Mann-Whitney U test, one-factor ANOVA test, and Spearman's test were used to determine the relationship between heparanase expression, microvessel density, and clinicopathological parameters. The expression of heparanase mRNA was detected in 20 of 40 (50%) endometrial cancers, and was significantly correlated with FIGO stage IIIc (p=0.0075), the presence of lymph-vascular space involvement (p=0.0041), lymph node metastasis (p=0.0049), and histological tumor grade (p=0.0030). Microvessel density was also associated with FIGO stage IIIc (p=0.027), the presence of lymph-vascular space involvement (p=0.001), lymph node metastasis (p=0.038), ovarian metastasis (p=0.030) and histological tumor grade (p=0.0030). Moreover, we found a strong positive correlation between heparanase expression and microvessel density (r2=0.475, p=0.0001). These results suggest that the expression of heparanase may influence different malignant behaviors in endometrial cancer. PMID- 12904691 TI - Medical management of endometriosis: novel targets and approaches towards the development of future treatment regimes. AB - Endometriosis remains an enigmatic disease in women of reproductive age. Current treatment regimes used to manage the disease are primarily designed to induce a hypoestrogenic state which leads to a reduction in disease and associated symptoms. While these regimes have been used with great success, there are still drawbacks and limitations to these types of therapies. In this review, we address the shortcomings of current drugs used to medically manage endometriosis and offer potential target areas which may be attractive alternatives to current therapies. Emphasis is placed upon the promising research using tumor necrosis factor inhibitors in this context. Further, we discuss the advantages of such therapies and offer experimental approaches towards the development and testing of these regimes. PMID- 12904692 TI - Different proliferative and apoptotic activity in peripheral versus central parts of human uterine leiomyomas. AB - OBJECTIVE: To determine whether there are differences in proliferative and apoptotic activity and in expression of sex steroid receptors between central and peripheral parts of human uterine leiomyomas in different menstrual cycle phases. METHODS: Biopsy specimens of the myometrium and peripheral and central parts of uterine leiomyomas were obtained from 15 women in the proliferative phase and 8 women in the secretory phase. Mitotic cells were detected by immunohistochemical staining for Ki67. Apoptotic cells were detected by the TUNEL method. Mitotic and apoptotic indexes were calculated. Tissue levels of oestrogen and progesterone receptors were measured using a commercial monoclonal receptor enzyme immunoassay kit. RESULTS: During the secretory phase the mitotic index was significantly higher in the peripheral than in the central parts of the leiomyomas. During the proliferative phase the apoptotic index was significantly higher in the peripheral compared with the central parts. These differences were not reflected by differences in receptor expression. CONCLUSIONS: The results of this study suggest that the growth of a human uterine leiomyoma mainly occurs in the peripheral parts of the tumour, during the secretory phase of the menstrual cycle. The findings emphasise the importance of being consistent when taking samples for research work. PMID- 12904693 TI - Detection of chronic endometritis by diagnostic hysteroscopy in asymptomatic infertile patients. AB - Chronic endometritis has been related to infertility and recurrent abortion. It is usually asymptomatic, and the diagnosis is rarely clinically suspected. We performed a prospective study to evaluate both the role of diagnostic hysteroscopy in the detection of chronic endometritis in infertile patients and Chlamydia trachomatis is a potential etiologic factor. Fifty consecutive patients who sought treatment for infertility in a tertiary academic hospital were submitted to diagnostic hysteroscopy and an endometrial biopsy for histopathological study and for diagnosis of C. trachomatis by polymerase chain reaction. The patients' mean age was 33.7 +/- (SD) 5.4 years, and the duration of the couples' infertility ranged from 1 to 18 years. The overall prevalence of chronic endometritis was 12% (6 patients). Among all patients, no cases of chlamydial infection were detected by polymerase chain reaction. In the detection of chronic endometritis, with 95% confidence intervals, the hysteroscopy sensitivity was 16.7% (range 0.9-63,5%), the specificity was 93.2% (range 80.3 98.2%), the positive predictive value was 25% (range 1.3-78.1%), and the negative predictive value was 89.1% (range 65.6-95.9%). These data suggest that hysteroscopy is not useful in the screening for chronic endometritis in asymptomatic infertile women. Further studies are needed to establish the etiology of endometritis in infertile patients. PMID- 12904694 TI - Tubo-ovarian abscess in older women: is the woman's age a risk factor for failed response to conservative treatment? AB - OBJECTIVE: To assess retrospectively if there is a trend of change in the epidemiology of tubo-ovarian abscess (TOA) and to define the epidemiologic, clinical and laboratory risk factors associated with failed response to conservative antibiotic therapy. STUDY DESIGN: The charts of 60 patients, admitted with clinically and sonographically diagnosed TOA between January 1995 and December 2000, were reviewed. On admission, all patients were treated with broad-spectrum antibiotics, and were divided into two groups according to the response to medical treatment. Forty-three patients, responding to antibiotic therapy, constituted group A, whereas 17 patients who did not respond and were further treated by sonographically guided drainage or surgery, were included in group B. The groups were compared with respect to patient characteristics, clinical and sonographic presentation, laboratory findings, duration of hospital stay and recovery rate of different pathogens from the endocervix. RESULTS: The mean age of patients responding to medical therapy (group A) was significantly lower (39.6 +/- 8.3) than the age of patients who did not respond (group B) (45.3 +/- 6.6) (p = 0.02). The gravidity as well as the percentage of menopausal women were significantly increased in group B (p = 0.03 and p = 0.02, respectively). There was a significantly lower incidence of previous pelvic surgery and past history of PID in group B versus group A (p = 0.02 and p = 0.03, respectively), yet the duration of IUD use was significantly prolonged in group B (p = 0.02). The size of TOA, evaluated clinically and sonographically, was significantly increased in group B compared with group A (p = 0.04 and p = 0.009, respectively). Pelvic peritonitis was observed in 40% of group B patients and in none of group A patients (p = 0.0001). The only laboratory finding differentiating between the two groups was the mean sedimentation rate, being significantly higher in group B than in group A (p = 0.0005). The time interval from admission to afebrile condition as well as the duration of hospitalization were significantly shorter in group A (6.1 +/- 2.3 days) than in group B (10.7 +/ 4.7 days) (p = 0.00003). The incidence of positive culture from the endocervix was significantly higher in group B than in group A (p = 0.007). One hundred percent of patients in group B versus 65.2% of patients in group A were treated with triple agent therapy (p = 0.007). CONCLUSION: The results of the current study strengthen our previous assumption that there is probably a new trend in the epidemiology of TOA, occurring in older women, who do not present the traditional risk factors for pelvic inflammatory disease and TOA. PMID- 12904696 TI - Abdominal wall and surgical scar endometriosis: results of magnetic resonance imaging. AB - Scar endometriosis is a rare disease which is difficult to diagnose. The symptoms are nonspecific, typically involving abdominal wall pain at the time of menstruation. Clinical examination may reveal a painful nodule, if the scar involved is located on the abdominal wall, but is normal, when the lesion is located on the uterine scar. Other means of investigation (transvaginal ultrasonography, computed tomography) may be useful in case of lesions on the abdominal wall, or if the nodule is large, but give no specific results. The diagnosis is frequently made only after excision of the lesion. We report here 4 patients operated for scar endometriosis (two abdominal and two uterine scars) for whom MRI had suggested the diagnosis. Thanks to its very high spatial resolution, MRI enables very small lesions to be detected and can distinguish the hemorrhagic signal of endometriotic lesions. Furthermore, it performs better than the CT scan in detecting the limits between muscles and abdominal subcutaneous tissues. PMID- 12904695 TI - Long-term follow-up (5-20 years) after uterine ventrosuspension for chronic pelvic pain and deep dyspareunia. AB - OBJECTIVE: To examine the long-term outcome of patients undergoing uterine ventrosuspension for chronic pelvic pain and deep dyspareunia. METHODS: Eighty two patients, presenting with retroverted uterus, pelvic pain and deep dyspareunia, underwent uterine ventrosuspension by laparotomy or laparoscopy during the period from 1981 to 1996. Only patients (56) with no pelvic pathologic condition, as identified intraoperatively, participated in the study. At the beginning of the year 2002, 41 out of 56 patients (73.2%) were attained by telephonic questionnaire, and they were asked to refer to the uterine ventrosuspension procedure. RESULTS: Patients' average age was 32.3 years, and the mean duration of preoperative symptoms was 3 years. The mean follow-up period was 12 (range 5-20) years. The initial improvement in preoperative symptoms was observed in 70.7% of patients, whereas the final improvement was observed in 46.3% of patients, with no significant difference comparing the approach of laparotomy versus laparoscopy. Hysterectomy after uterine suspension was performed in 12.1% of patients (4.3% after laparotomy and 22.2% after laparoscopy). Twenty-three patients (56.1%) would recommend this operative procedure to others. CONCLUSION: The long-term outcome of patients undergoing uterine ventrosuspesnion approximates to 50% success rate, with no significant difference comparing the approach of laparotomy versus laparoscopy. PMID- 12904697 TI - mRNA-expression of often used house-keeping genes and the relation between RNA and DNA are sex steroid-dependent parameters in human myometrium and fibroids. AB - The content of RNA and DNA in human myometrium and fibroids obtained at different endocrine conditions varied, with the highest RNA/DNA ratio in tissues from pregnant patients, intermediate ratios in women during the menstrual cycle and the lowest in tissues from postmenopausal and GnRHa-treated patients. mRNA expression of two house-keeping genes, gamma-actin and GAPDH, was highest in myometrium from pregnant women, intermediate in untreated women of fertile age and lowest in tissues from GnRHa-treated and postmenopausal women. To control for degradation of nucleic acids when measuring mRNA expression, we suggest additional analysis of gene(s), where the expression pattern is known, and that expression, whenever possible, is related to DNA, which is a more stable parameter than RNA and total nucleic acids, when there are differences in proliferation between tissues and/or groups of patients. PMID- 12904699 TI - A probability model for ultrasound estimation of bladder volume in the diagnosis of female urinary retention. AB - OBJECTIVE: The aim of this study was to evaluate a probability model for ultrasound estimation of bladder volume in the diagnosis of female urinary retention, which is a common condition in the postpartum period. METHODS: A total of 53 women with vaginal delivery were recruited on postpartum day 1, and their bladder volumes were first estimated by ultrasound. Immediately after the ultrasound scans the bladders were catheterized for the true bladder volumes. The ultrasound-estimated bladder volumes were then compared with the catheterized bladder volumes. The data were first tested for non-normality, and then logarithmic transformed in order to conform to normality. A probability model was built using the logarithmic-transformed data, based on the following: an arbitrary cut-off value of 150 ml for urinary retention, linear regression analysis, and paired difference analysis. RESULTS: There was a linear relationship between the ultrasound-estimated and catheterized bladder volumes, which can be represented by the equation: log(10) (Vc) = 0.2959 + 0.8853 log(10) (Vu), where Vc = catheterized bladder volume, and Vu = ultrasound-estimated bladder volume. A normogram was constructed, and the probability of urinary retention presented. CONCLUSIONS: The results of this study showed that our method and statistics are logical by which the probability model of urinary retention is precise enough to support clinical decision-making. PMID- 12904698 TI - Laparoscopy-assisted vaginal hysterectomy clinical pathway. A multivariate analysis of impact on costs and quality of care. AB - Numerous studies have demonstrated that a well-designed clinical pathway is an effective means of sustaining quality while controlling costs in the management of certain disease entities. We evaluated the impact that cost and medical quality have on the implementation of a clinical pathway for laparoscopy-assisted vaginal hysterectomy (LAVH). This retrospective study involved a sample of 124 patients who underwent LAVH in a medical center in central Taiwan. Patients were divided into two groups on the basis of whether they received treatment before or after implementation of the LAVH clinical pathway. The preclinical pathway group was comprised of 40 patients who underwent LAVH before clinical pathway implementation (May-December 1997). The clinical pathway group included 84 patients who underwent LAVH after implementation of the clinical pathway (January 1998-March 1999). In order to study the impact of the LAVH clinical pathway, patient characteristics were controlled by multiple linear regression. The results showed a significant reduction in cost, average length of hospital stay, and average duration of surgery and anesthesia (p < 0.01). Dependent nominal variables for clinical indicators like postoperative intravenous fluid and injection of antibiotics 48 h after surgery, and complications were analyzed by a logistic regression model. The results noted better control of antibiotic intravenous injection 48 h after surgery in the clinical pathway group (p = 0.03). The other indicators included delay of operation day, blood transfusion, patient mortality, and patients readmitted within 2 weeks. There was one operation day delay and one readmission within 2 weeks of discharge in the preclinical pathway group. Based on our results, the implementation of a clinical pathway for LAVH contains cost while maintaining quality of care, especially when the medical fees are paid under the case payment system. PMID- 12904700 TI - Serial transvaginal sonography of a case of complete hydatidiform mole. AB - Sonography has achieved a position of preeminence in the diagnostic evaluation of molar gestation. However, little information is available about the serial change of the transvaginal sonographic features of molar pregnancy. We describe a case of complete hydatidiform mole, and present serial transvaginal views taken during the early gestational period. PMID- 12904701 TI - Disorders of pulmonary circulation. PMID- 12904702 TI - The role of low-molecular-weight heparins in the prevention and treatment of venous thromboembolism in cancer patients. AB - Accumulating evidence suggests that low-molecular-weight heparins are the drug of choice for the prevention and treatment of venous thromboembolism in patients with cancer. For prophylaxis in the surgical setting, once-daily subcutaneous injections of low-molecular-weight heparin are as effective and safe as multiple doses of unfractionated heparin. Extending prophylaxis with low-molecular-weight heparins beyond hospitalization was recently found to reduce safely the risk of postoperative thrombosis after abdominal surgery for cancer. For the long-term treatment of deep vein thrombosis and in select patients with pulmonary embolism, recently completed clinical trials have shown that secondary prophylaxis with low molecular-weight heparin is feasible and more effective than oral anticoagulant therapy in preventing recurrent venous thromboembolism in cancer patients. There is also evidence that low-molecular-weight heparins are effective in cancer patients who develop recurrent thrombosis while on warfarin therapy. Lastly, the potential antineoplastic effects of low-molecular-weight heparins make these agents an attractive option in patients with cancer. Although the management of cancer patients with venous thromboembolism remains challenging, low-molecular weight heparins have simplified and improved the prevention and treatment of venous thromboembolism in these high-risk patients. PMID- 12904704 TI - Central line thrombosis in patients with cancer. AB - Central venous catheters are used frequently to care for patients with cancer and often serve as long-term venous access. Catheter-related central venous thrombosis is a serious and common complication in these patients. The incidence of this event ranges from 2 to 40%. Potential risk factors are catheter position, size of the catheter, and perhaps site of insertion. The diagnosis of catheter associated deep venous thrombosis may be difficult. Doppler ultrasound has a lower accuracy in this setting than it does in symptomatic lower extremity venous thrombosis. No clinical management studies have validated the practice of withholding anticoagulant therapy in patients with a negative Doppler ultrasound. The practice of prophylaxis with low-dose warfarin or low-molecular-weight heparin has been recommended, although results from recent studies have called this into question. Larger, prospective, randomized trials with a uniform population of patients or stratification of risk factors will be essential to address this issue further. PMID- 12904703 TI - Venous thromboembolism in laparoscopic surgery. AB - Currently there are limited randomized data regarding thromboprophylaxis in laparoscopic surgery. The aim of this article is to identify principles to guide safe practice with regard to prevention of venous thromboembolism. With the exception of laparoscopic cholecystectomy, there are no prospective, randomized studies comparing the incidence of venous thromboembolism between a conventional procedure and a laparoscopic procedure for the same operation. Surveys of surgical practice indicate that policies for venous thromboembolism prophylaxis in laparoscopic surgery are generally the same as those for conventional surgery. The increasing use of a minimal access approach for a variety of abdominal, pelvic, and thoracic procedures demands further prospective, randomized studies in this area. Current guidelines endorsed by The Society of American Gastrointestinal Endoscopic Surgeons recommend following the adoption of protocols used in conventional surgery for the equivalent laparoscopic operation, and the European Association for Endoscopic Surgery has recommended the use of intraoperative intermittent pneumatic compression of the lower extremities for all prolonged laparoscopic procedures. There is only limited evidence to support these recommendations. Venous thromboembolism is an important and preventable complication in surgical patients. Evidence does not exist to consider laparoscopic surgery patients to be at a substantially lower risk for venous thromboembolism than those undergoing conventional procedures. Currently there is a lack of data regarding the prophylaxis against venous thromboembolism in laparoscopic surgery, and the practice is thus opinion based. The authors recommend that the use of standard prophylactic regimens tailored to specific patient populations for conventional operations be adopted for laparoscopic surgery until prospective data are available. PMID- 12904705 TI - CT for the diagnosis of venous thromboembolic disease. AB - Spiral CT, venous ultrasound, ventilation-perfusion scanning, and D-dimer tests are routinely used in the diagnosis of venous thromboembolic disease. Advances in multidetector spiral CT and the combination of CT pulmonary angiography and venography as a one-step evaluation of venous thromboembolic disease have markedly improved detection of subsegmental emboli and deep pelvic venous thrombi and decreased the role of conventional pulmonary angiography. As CT has improved, many have questioned what "gold standard" CT should be compared with. Recent clinical outcome studies suggest that CT results are as good as pulmonary angiography and conventional imaging algorithms. Because the sensitivity of CT pulmonary angiography now exceeds 85%, it also appears that it is also more cost effective than other diagnostic approaches. PMID- 12904706 TI - Troponin levels as a guide to treatment of pulmonary embolism. AB - Right ventricular dysfunction in hemodynamically stable patients with acute pulmonary embolism may be a harbinger of adverse outcomes and may potentially result in the early use of thrombolytic therapy. Risk stratification of these patients is an area of recent and intense investigation with a focus on the assessment of right ventricular function after the embolic event. Echocardiography has been used to identify right ventricular dysfunction but is potentially hampered by a number of limitations. With the onset of right ventricular dilation and possible ischemia in acute pulmonary embolism, elevated serum troponins may be an early and reliable marker of right ventricular dysfunction. In acute pulmonary embolism, both right ventricular dysfunction by echocardiogram and elevated troponin levels have been shown to predict an adverse outcome. Therefore, serum troponin levels should help stratify patients with submassive acute pulmonary embolism into a group in which aggressive medical or surgical intervention would be considered. PMID- 12904707 TI - Dyslipidemia, statins, and venous thromboembolism: a potential risk factor and a potential treatment. AB - The optimal drug for the prevention of venous thromboembolism is one that is efficacious, associated with minimal bleeding risk, and easy to administer. Statins fulfill the latter two criteria, but their efficacy remains unproved. By examining the association between dyslipidemia and venous thromboembolism, as well as the evidence that statins might prevent venous thromboembolism, there may be a new rationale for the use of this class of drugs. There may be a common link between arterial and venous thrombosis. Dyslipidemia may be one of the many systemic factors associated not only with arterial thrombosis, but with venous thromboembolism as well. This may occur through the effects of circulating lipid molecules on the vascular endothelium, platelet function, and coagulation factors. By impeding these mechanisms, statins may be protective against venous thrombosis, but epidemiologic studies are few in number, and no randomized clinical trials have been conducted. Better epidemiologic evidence is required to establish whether dyslipidemia is a risk factor for venous thromboembolism. If future observational studies can demonstrate that statins are associated with a lower risk of venous thromboembolism, then consideration should be given to conducting a randomized clinical trial comparing statins with placebo for the prevention of venous thromboembolism. Until then, the efficacy of statins for the prevention or treatment of venous thromboembolism remains uncertain. PMID- 12904708 TI - Hypercoagulable states and central retinal vein occlusion. AB - Central retinal vein occlusion is a common cause of permanent visual loss. Work up and laboratory evaluation of patients requires the clinician to rule out hypertension, diabetes, hyperlipidemia, and glaucoma. Patients without an identifiable risk factor are often subject to extensive testing for primary and secondary thrombophilias. The purpose this paper is to review the literature to determine which of these tests is associated with central retinal vein occlusion. Antiphospholipid antibodies and elevated plasma homocysteine levels appear to be the tests associated most commonly in patients with central retinal vein occlusion in most controlled studies. Primary thrombophilias are found rarely when screening patients with central retinal vein occlusion. Extensive testing for thrombophilias is not warranted in the vast majority of patients with central retinal vein occlusion. Older patients with any of the common vascular risk factors do not require thrombophilic screening. By carefully selecting the patients who are evaluated for thrombophilias, the likelihood of finding true positive tests is increased. PMID- 12904709 TI - Superficial vein thrombosis: risk factors, diagnosis, and treatment. AB - Superficial vein thrombosis (SVT) risk factors are close to those of venous thromboembolism (VTE). Diagnosis is made in a clinical setting but ultrasonography is useful to eliminate concomitant deep vein thrombosis (DVT). For SVT of the lower limbs, which is the main location, varicose veins represent the principal cause but underlying conditions (e.g.: autoimmune diseases, malignancy or thrombophilia) must be sought in idiopathic, migrant or recurrent SVT and in the absence of varicose veins. Concomitant DVT and pulmonary embolism can occur in approximately 15% and 5% respectively. Historical treatments consist of anti-inflammatory agents plus elastic stockings and, in case of varicose veins, thrombectomy and stripping. Other treatments (anticoagulants, vein ligation) were assessed to limit the VTE risk. A one-month prophylactic dose of low molecular weight heparin plus elastic stockings could be the appropriate strategy in most cases. Other studies are needed before definitive conclusions can be drawn. PMID- 12904710 TI - Cancer in interstitial pulmonary fibrosis and sarcoidosis. AB - The occurrence of lung cancer in patients with diffuse interstitial pulmonary disease is well-known. The incidence, however, varies and ranges from 9.8 to 38%. The pathogenesis of lung cancer in pulmonary fibrosis is unclear, but genetic as well as environmental factors seem to be involved. PMID- 12904711 TI - Bronchoscopy in sarcoidosis: diagnostic and therapeutic interventions. AB - Flexible bronchoscopy has revolutionized the evaluation of patients with suspected sarcoidosis and the treatment of sarcoid patients with significant endobronchial disease. The authors explore the diagnostic and therapeutic utility of flexible bronchoscopy by reviewing the pertinent literature with a special interest in recent studies. Bronchoscopy allows multiple diagnostic modalities in suspected sarcoidosis. Recent studies show sometimes surprising results, and the authors review the additive contributions of transbronchial lung biopsy, endobronchial biopsy, transbronchial needle aspiration, and bronchoalveolar lavage to diagnose sarcoidosis. New data specifically show the additive benefit of routine endobronchial biopsy and transbronchial needle aspiration to traditional transbronchial biopsy specimens. In addition, the techniques have been optimized via recent studies and these results are discussed. Endobronchial therapy is reviewed with the recent findings of the superiority of balloon bronchoplasty. Flexible bronchoscopy has a very high diagnostic yield in all stages of suspected sarcoidosis. Transbronchial lung biopsy and endobronchial biopsy should be used routinely, and transbronchial needle aspiration should be considered in cases of significant adenopathy. Bronchoalveolar lavage should be used routinely to exclude alternative diagnoses. Therapeutic bronchoscopy is rarely needed, but when necessary the authors' procedure of choice is bronchoscopic balloon dilatation. PMID- 12904712 TI - Role of exercise evaluation in restrictive lung disease: new insights between March 2001 and February 2003. AB - Restrictive lung disease is often first detected when patients complain of dyspnea on exertion. Many forms of exercise testing are available, from simple hallway oximetry to the more formal and more complex cardiopulmonary exercise test. Although the use of exercise for diagnosis, treatment, and predicting outcomes is largely understudied in this population, it has recently been shown to be of value in some settings. Exercise testing may be a valuable diagnostic tool in determining the extent of lung disease in sarcoidosis. Medinger et al. reported that the symptom-limited exercise test detected pulmonary dysfunction earlier than history, physical examination, chest radiography, and spirometry alone. Furthermore, Delobbe et al. noted that in patients with biopsy-proved sarcoidosis, cardiopulmonary exercise testing was a more sensitive indicator of early lung disease than pulmonary function tests. The American College of Chest Physicians/American Thoracic Society have published an updated consensus statement for cardiopulmonary exercise testing. Christensen et al. reported that patients with restrictive lung disease may be at risk for hypoxemia with light exercise while on an airplane, and suggest that these patients be considered for in-flight oxygen therapy. Lastly, Herridge and the Canadian Critical Care Trial Group used the 6-minute walk test to prove that survivors of acute respiratory distress syndrome have significant functional limitation 1 year after discharge from the intensive care unit largely secondary to neuromuscular sequelae. Exercise testing appears to be a valuable tool in evaluating, treating, and predicting outcomes in patients with restrictive lung disease. Further study will help to support its use in other restrictive lung diseases. PMID- 12904713 TI - Recent advances in the diagnosis and management of nonspecific interstitial pneumonia. AB - Idiopathic interstitial pneumonia is a heterogeneous group of diseases. Recently, the importance of a detailed differentiation of nonspecific interstitial pneumonia from other idiopathic interstitial pneumonias has been demonstrated. Most critical appeared to be a proper classification based on clinical presentation, high-resolution CT findings, bronchoalveolar lavage fluid cell findings, and histopathology. This classification may guide the most appropriate therapeutic approach and has significant implications regarding prognosis. Recent advances in the diagnosis and management of nonspecific interstitial pneumonia, cellular and fibrosing variants, are discussed. PMID- 12904714 TI - Is bronchoalveolar lavage obsolete in the diagnosis of interstitial lung disease? AB - This review considers the literature on bronchoalveolar lavage in the diagnosis of interstitial lung disease published during the last 12 months with the aim of clarifying the role bronchoalveolar lavage can have in diagnostic work-up on the basis of current knowledge and expert opinion. Recent research findings with possible future clinical applications are presented. Various information, useful for research and clinical applications, can be obtained from performing bronchoalveolar lavage in patients with interstitial lung diseases. Indeed, evaluation of cell pattern associated with cell phenotype is used widely in clinical practice to distinguish the various forms and may be of diagnostic value in some interstitial lung diseases, as already known, when the clinical picture is compatible. Bronchoalveolar lavage may also be complementary to high resolution CT or at least useful for diagnosis by exclusion. A major advance in the last year is recognition of a role for bronchoalveolar lavage in the diagnostic workup of idiopathic interstitial pneumonias, albeit as an auxiliary procedure. It may be useful to exclude infections and tumors, may help to decide whether to do surgical biopsy, and may aid in distinguishing different forms of interstitial lung disease. Although it is not diagnostic for idiopathic interstitial pneumonias, in the presence of cell patterns considered "typical" of the various forms, it can support clinical diagnosis in the absence of biopsy. Because further studies following standardized protocols and guidelines will presumably find new parameters for bronchoalveolar lavage in the diagnostics of interstitial lung diseases, it would be a mistake to consider bronchoalveolar lavage an obsolete tool. PMID- 12904715 TI - Management of pulmonary hypertension resulting from interstitial lung disease. AB - Pulmonary hypertension affects right ventricular function by imposing an afterload on the right ventricle and, therefore, as pulmonary hypertension worsens, it ultimately reduces cardiac output. It is clear that in interstitial lung disease, progression of the underlying process with the loss of lung parenchyma contributes substantially to outcome. However, the additional burden of pulmonary hypertension often results in rapid deterioration. Pulmonary hypertension may be treatable and thus may enable the patient to survive until other therapy is effective. Until recently, part of the hesitancy in treating pulmonary hypertension in patients with secondary pulmonary hypertension, has been the limited availability of oral or intravenous drugs that affect the pulmonary circuit without causing excessive systemic vasodilatation. The current drugs available for the treatment of pulmonary hypertension, including prostacyclin and endothelin receptor blockers, have been limited by their high costs and, in the case of prostacyclin, the drug delivery systems. Nitric oxide, which has been used both acutely and chronically for pulmonary hypertension in a variety of diseases, also has limited availability. The role of sildenafil and inhaled prostacyclin remains to be evaluated. The use of these agents in the setting of interstitial lung disease needs to be studied, including combination therapy and early initiation of therapy. Their effect on tissue damage, fibrosis, and inflammation needs to be assessed as well as their effect on the arteriopathy and their potential to cause ventilation-perfusion (V/Q) mismatching. There now appears to be evidence to support treating pulmonary hypertension in patients with interstitial lung disease, with a potential for clinical benefit and a molecular basis for doing it. PMID- 12904716 TI - High-resolution CT: what is it good for in pulmonary fibrosis? AB - For many radiologists and clinicians, high-resolution CT has become a useful tool in the diagnosis of diffuse lung disease, particularly in the diagnosis of idiopathic pulmonary fibrosis. This paper reviews the most recent literature on the usefulness of high-resolution CT in diagnosing, determining the subsequent prognosis, and guiding the care of patients with lung disease. Idiopathic lung diseases, collagen vascular diseases, and the role of lung biopsy in diffuse lung disease are addressed. PMID- 12904717 TI - New observations on pulmonary function changes in sarcoidosis. AB - Respiratory tract involvement occurs at some time during the course of most patients with sarcoidosis. There are many articles on lung function impairment in sarcoidosis, but the definite, unique guideline considering the correlation between the clinical, radiologic, and biochemical findings of the disease is lacking. During the last few years, most of the literature has focused on follow up studies, presenting primarily the effects of the treatment on lung function impairment in patients with sarcoidosis. This study focuses on airflow impairment, diffusion impairment, and bronchial hyperresponsiveness in sarcoidosis. The topic has already been explored, but this time the authors emphasize lung function impairment and its correlation with the course of the disease. Considering the course of sarcoidosis, a significantly higher number of patients with diffusion impairment have the chronic form of the disease. The bronchial challenge test (BCT) was performed in patients with chronic sarcoidosis at the time they experienced relapse of disease activity. A high number of patients with positive BCT is significant in light of their disease. Further analyses, not only functional, but immunologic, are necessary to examine the potential correlation between positive BCT and the activity of sarcoidosis. PMID- 12904719 TI - Odd woman out. PMID- 12904718 TI - Assisting living: where are the RNs? PMID- 12904722 TI - Reflections: Back from remission. PMID- 12904723 TI - Policy perspectives: Assisted living and the role of nursing. PMID- 12904724 TI - Stress urinary incontinence in women. PMID- 12904725 TI - Emergency: Monophasic and biphasic defibrillators. PMID- 12904726 TI - Addressing inadequate pain relief. PMID- 12904730 TI - Solidarity. PMID- 12904731 TI - Not so fast! PMID- 12904732 TI - Community rebirth in Cornwall. PMID- 12904733 TI - Behavioral counseling in primary care to promote a healthy diet: recommendations and rationale. PMID- 12904736 TI - Taking risks, tapping creativity. PMID- 12904737 TI - Barriers to reporting a workplace injury. PMID- 12904738 TI - Imported plague--New York City, 2002. AB - On November 1, 2002, a married couple traveled from Santa Fe County, New Mexico, to New York City (NYC), where they both became ill with fever and unilateral inguinal adenopathy; bubonic plague (Yersinia pestis) was diagnosed subsequently. This report summarizes the clinical and public health investigation of these cases and underscores the importance of rapid diagnosis and communication among health-care providers, public health agencies, and the public when patients seek medical attention for an illness that might be caused by an agent of terrorism. PMID- 12904739 TI - National, state, and urban area vaccination levels among children aged 19-35 months--United States, 2002. AB - Each annual birth cohort in the United States comprises approximately four million infants. Maintaining the gains in childhood vaccination coverage achieved during the 1990s among these children poses an ongoing challenge for public health. The National Immunization Survey (NIS) provides annual estimates of vaccination coverage among children aged 19-35 months for each of the 50 states and 28 selected urban areas. This report presents NIS findings for 2002, which indicate a marked nationwide increase in coverage with >/=1 dose of varicella vaccine (VAR), substantial uptake for >/=3 doses of pneumococcal conjugate vaccine (PCV), generally steady coverage levels for other vaccines nationwide, and continued wide variability in coverage among the states and selected urban areas. PMID- 12904740 TI - Vaccination services in postwar Iraq, May 2003. AB - In the aftermath of the war in Iraq, widespread looting and intentional damage to government facilities resulted in the interruption of public services and utilities. Basic communications were disrupted nationally. Public health headquarters, clinics, and laboratories were damaged, records were ruined, and equipment was stolen. Because travel often was difficult and dangerous, Coalition forces received numerous requests from hospital directors for armed security, and many health-care workers reportedly feared either to commute to their worksites or to remain after dark (D. Simpson, M.D., Coalition Provisional Authority [CPA]'s Ministry of Health Team, personal correspondence, 2003). Public health employees who were able to continue their work went unpaid for several weeks. As a result, throughout Iraq, core public health services (e.g., vaccination services, vectorborne disease control, and the Tuberculosis Directly Observed Therapy program) were disrupted. In addition, severe health hazards caused by damaged water and sanitation systems were added to an already compromised and deteriorating health-care system. This report assesses the cumulative impact of these conditions on vaccination services in postwar Iraq, including the subsequent loss of staff, facilities, and equipment. Because vaccinations in Iraq are available only through the national system of primary health-care centers (PHCCs), this assessment can help address comparable problems experienced by other programs offered through Iraq's PHCCs, guide subsequent emergency responses to vaccine shortages, and provide a preliminary gauge of the status of preventive health-care infrastructure and services to children in Iraq. PMID- 12904741 TI - Update: adverse event data and revised American Thoracic Society/CDC recommendations against the use of rifampin and pyrazinamide for treatment of latent tuberculosis infection--United States, 2003. AB - CDC has reported previously surveillance data of severe liver injury in patients treated for latent tuberculosis infection (LTBI) with a daily and twice-weekly 2 month regimen of rifampin with pyrazinamide (RZ). On the basis of these initial reports, CDC cautioned clinicians in the use of this therapy with advised additional monitoring. To estimate the incidence of RZ-associated severe liver injury and provide more precise data to guide treatment for LTBI, CDC collected data from cohorts of patients in the United States who received RZ for the treatment of LTBI during January 2000-June 2002 and for whom data were reported to CDC through June 6, 2003. This report summarizes the analysis, which found high rates of hospitalization and death from liver injury associated with the use of RZ. On the basis of these findings, the American Thoracic Society (ATS) and CDC now recommend that this regimen should generally not be offered to persons with LTBI. The revised ATS/CDC recommendations described in this report have been endorsed by the Infectious Diseases Society of America (IDSA). Clinicians are advised to use the recommended alternative regimens for the treatment of LTBI. Rifampin and pyrazinamide (PZA) should continue to be administered in multidrug regimens for the treatment of persons with active tuberculosis (TB) disease. PMID- 12904744 TI - In defence of DARPA. PMID- 12904745 TI - Hubble versus the future. PMID- 12904746 TI - Terrorist betting leaves defence agency fighting for autonomy. PMID- 12904747 TI - Fast vaccine offers hope in battle with Ebola. PMID- 12904748 TI - Czech stem-cell work heightens calls for EU ruling. PMID- 12904749 TI - NASA under pressure to extend Hubble's life. PMID- 12904750 TI - Ecological advice sparks sea change in judicial opinion. PMID- 12904751 TI - WHO prepares for final push to rid the world of polio. PMID- 12904752 TI - Piglets add some colour to transgenic story. PMID- 12904753 TI - Red tape frustrates Europe's fund-seekers. PMID- 12904754 TI - Companies vie to put all your genes on a chip. PMID- 12904756 TI - Venice floods: SAVE OUR CITY! PMID- 12904757 TI - DNA microarrays: vital statistics. PMID- 12904758 TI - Natural decaf could brew trouble for farmers. PMID- 12904759 TI - Getting to the heart of transpiration in plants. PMID- 12904760 TI - Flawed science underlies laws on transgenic crops. PMID- 12904766 TI - Science in culture. PMID- 12904767 TI - Genetic engineering: unnatural selection. PMID- 12904769 TI - Nanotechnology: a barrier falls. PMID- 12904768 TI - Animal locomotion: how to walk on water. PMID- 12904770 TI - Atmospheric science: African dust in Florida clouds. PMID- 12904772 TI - Superconductivity: lifting the gossamer veil. PMID- 12904771 TI - Signal transduction: life on Mars, cellularly speaking. PMID- 12904774 TI - Neurobiology: a thorny issue. PMID- 12904775 TI - Astronomy: an elementary puzzle. PMID- 12904777 TI - Particle physics: antimatter matters. PMID- 12904778 TI - Pregnancy: a cloned horse born to its dam twin. AB - Several animal species, including sheep, mice, cattle, goats, rabbits, cats, pigs and, more recently, mules have been reproduced by somatic cell cloning, with the offspring being a genetic copy of the animal donor of the nuclear material used for transfer into an enucleated oocyte. Here we use this technology to clone an adult horse and show that it is possible to establish a viable, full-term pregnancy in which the surrogate mother is also the nuclear donor. The cloned offspring is therefore genetically identical to the mare who carried it, challenging the idea that maternal immunological recognition of fetal antigens influences the well-being of the fetus and the outcome of the pregnancy. PMID- 12904779 TI - Lithium-ion batteries: runaway risk of forming toxic compounds. AB - Lithium-ion batteries are stabilized by an ultrathin protective film that is 10 50 nanometers thick and coats both electrodes. Here we artificially simulate the 'thermal-runaway' conditions that would arise should this coating be destroyed, which could happen in a battery large enough to overheat beyond 80 degrees C. We find that under these conditions the reaction of the battery electrolyte with the material of the unprotected positive electrode results in the formation of toxic fluoro-organic compounds. Although not a concern for the small units used in today's portable devices, this unexpected chemical hazard should be taken into account as larger and larger lithium-ion batteries are developed, for example for incorporation into electric-powered vehicles. PMID- 12904780 TI - Palaeontology: spider-web silk from the Early Cretaceous. AB - The use of viscid silk in aerial webs as a means to capture prey was a key innovation of araneoid spiders and has contributed largely to their ecological success. Here I describe a single silk thread from a spider's web that bears glue droplets and has been preserved in Lebanese amber from the Early Cretaceous period for about 130 million years. This specimen not only demonstrates the antiquity of viscid silk and of the spider superfamily Araneoidea, but is also some 90 million years older than the oldest viscid spider thread previously reported in Baltic amber from the Eocene epoch. PMID- 12904781 TI - Insect signalling: components of giant hornet alarm pheromone. AB - Up to 74 people die each year in Japan after being stung by Hymenopteran insects, with hornets (Vespa spp.) being among the worst offenders. Here we identify a volatile, multi-component alarm pheromone in the venom of the world's largest hornet, V. mandarinia, and use field bioassays to show that 2-pentanol is its principal active component, and that 3-methyl-1-butanol and 1-methylbutyl 3 methylbutanoate act synergistically with it. The compound 1-methylbutyl 3 methylbutanoate, which may also be a foraging-site-marking pheromone, elicits a strong defensive reaction in the sympatric prey hornet V. simillima xanthoptera. As these chemicals are sometimes used in food flavourings and as fragrances in cosmetics, it is possible that they might provoke a seemingly unwarranted hornet attack on humans. PMID- 12904782 TI - Optics (communication arising): mechanism for 'superluminal' tunnelling. AB - In their discussion of a mechanism that I proposed to explain the apparent superluminal (that is, faster than light) tunnelling of light pulses observed in photonic barrier experiments, Buttiker and Washburn used an old 'reshaping' argument that is at variance with my model and is not supported by the bulk of the experimental tunnelling evidence. The mechanism I proposed agrees with experiment and resolves a long-standing paradox - namely, the lack of dependence of tunnelling time on barrier length for thick barriers (the Hartman effect). PMID- 12904784 TI - The evolution of comets in the Oort cloud and Kuiper belt. AB - Comets are remnants from the time when the outer planets formed, approximately 4 4.5 billion years ago. They have been in storage since then in the Oort cloud and Kuiper belt-distant regions that are so cold and sparsely populated that it was long thought that comets approaching the Sun were pristine samples from the time of Solar System formation. It is now recognized, however, that a variety of subtle but important evolutionary mechanisms operate on comets during their long storage, so they can no longer be regarded as wholly pristine. PMID- 12904785 TI - Complete atomic model of the bacterial flagellar filament by electron cryomicroscopy. AB - The bacterial flagellar filament is a helical propeller for bacterial locomotion. It is a helical assembly of a single protein, flagellin, and its tubular structure is formed by 11 protofilaments in two distinct conformations, L- and R type, for supercoiling. The X-ray crystal structure of a flagellin fragment lacking about 100 terminal residues revealed the protofilament structure, but the full filament structure is still essential for understanding the mechanism of supercoiling and polymerization. Here we report a complete atomic model of the R type filament by electron cryomicroscopy. A density map obtained from image data up to 4 A resolution shows the feature of alpha-helical backbone and some large side chains. The atomic model built on the map reveals intricate molecular packing and an alpha-helical coiled coil formed by the terminal chains in the inner core of the filament, with its intersubunit hydrophobic interactions having an important role in stabilizing the filament. PMID- 12904786 TI - An asymptotic-giant-branch star in the progenitor system of a type Ia supernova. AB - Stars that explode as supernovae come in two main classes. A type Ia supernova is recognized by the absence of hydrogen and the presence of elements such as silicon and sulphur in its spectrum; this class of supernova is thought to produce the majority of iron-peak elements in the Universe. They are also used as precise 'standard candles' to measure the distances to galaxies. While there is general agreement that a type Ia supernova is produced by an exploding white dwarf star, no progenitor system has ever been directly observed. Significant effort has gone into searching for circumstellar material to help discriminate between the possible kinds of progenitor systems, but no such material has hitherto been found associated with a type Ia supernova. Here we report the presence of strong hydrogen emission associated with the type Ia supernova SN2002ic, indicating the presence of large amounts of circumstellar material. We infer from this that the progenitor system contained a massive asymptotic-giant branch star that lost several solar masses of hydrogen-rich gas before the supernova explosion. PMID- 12904787 TI - Ballistic carbon nanotube field-effect transistors. AB - A common feature of the single-walled carbon-nanotube field-effect transistors fabricated to date has been the presence of a Schottky barrier at the nanotube- metal junctions. These energy barriers severely limit transistor conductance in the 'ON' state, and reduce the current delivery capability--a key determinant of device performance. Here we show that contacting semiconducting single-walled nanotubes by palladium, a noble metal with high work function and good wetting interactions with nanotubes, greatly reduces or eliminates the barriers for transport through the valence band of nanotubes. In situ modification of the electrode work function by hydrogen is carried out to shed light on the nature of the contacts. With Pd contacts, the 'ON' states of semiconducting nanotubes can behave like ohmically contacted ballistic metallic tubes, exhibiting room temperature conductance near the ballistic transport limit of 4e(2)/h (refs 4-6), high current-carrying capability (approximately 25 micro A per tube), and Fabry Perot interferences at low temperatures. Under high voltage operation, the current saturation appears to be set by backscattering of the charge carriers by optical phonons. High-performance ballistic nanotube field-effect transistors with zero or slightly negative Schottky barriers are thus realized. PMID- 12904788 TI - Low-loss hollow-core silica/air photonic bandgap fibre. AB - Photonic bandgap structures use the principle of interference to reflect radiation. Reflection from photonic bandgap structures has been demonstrated in one, two and three dimensions and various applications have been proposed. Early work in hollow-core photonic bandgap fibre technology used a hexagonal structure surrounding the air core; this fibre was the first demonstration of light guided inside an air core of a photonic bandgap fibre. The potential benefits of guiding light in air derive from lower Rayleigh scattering, lower nonlinearity and lower transmission loss compared to conventional waveguides. In addition, these fibres offer a new platform for studying nonlinear optics in gases. Owing largely to challenges in fabrication, the early air-core fibres were only available in short lengths, and so systematic studies of loss were not possible. More recently, longer lengths of fibre have become available with reported losses of 1,000 dB km(-1). We report here the fabrication and characterization of long lengths of low attenuation photonic bandgap fibre. Attenuation of less than 30 dB km(-1) over a wide transmission window is observed with minimum loss of 13 dB km(-1) at 1,500 nm, measured on 100 m of fibre. Coupling between surface and core modes of the structure is identified as an important contributor to transmission loss in hollow-core photonic bandgap fibres. PMID- 12904789 TI - Unusually large earthquakes inferred from tsunami deposits along the Kuril trench. AB - The Pacific plate converges with northeastern Eurasia at a rate of 8-9 m per century along the Kamchatka, Kuril and Japan trenches. Along the southern Kuril trench, which faces the Japanese island of Hokkaido, this fast subduction has recurrently generated earthquakes with magnitudes of up to approximately 8 over the past two centuries. These historical events, on rupture segments 100-200 km long, have been considered characteristic of Hokkaido's plate-boundary earthquakes. But here we use deposits of prehistoric tsunamis to infer the infrequent occurrence of larger earthquakes generated from longer ruptures. Many of these tsunami deposits form sheets of sand that extend kilometres inland from the deposits of historical tsunamis. Stratigraphic series of extensive sand sheets, intercalated with dated volcanic-ash layers, show that such unusually large tsunamis occurred about every 500 years on average over the past 2,000 7,000 years, most recently approximately 350 years ago. Numerical simulations of these tsunamis are best explained by earthquakes that individually rupture multiple segments along the southern Kuril trench. We infer that such multi segment earthquakes persistently recur among a larger number of single-segment events. PMID- 12904790 TI - The hydrodynamics of water strider locomotion. AB - Water striders Gerridae are insects of characteristic length 1 cm and weight 10 dynes that reside on the surface of ponds, rivers, and the open ocean. Their weight is supported by the surface tension force generated by curvature of the free surface, and they propel themselves by driving their central pair of hydrophobic legs in a sculling motion. Previous investigators have assumed that the hydrodynamic propulsion of the water strider relies on momentum transfer by surface waves. This assumption leads to Denny's paradox: infant water striders, whose legs are too slow to generate waves, should be incapable of propelling themselves along the surface. We here resolve this paradox through reporting the results of high-speed video and particle-tracking studies. Experiments reveal that the strider transfers momentum to the underlying fluid not primarily through capillary waves, but rather through hemispherical vortices shed by its driving legs. This insight guided us in constructing a self-contained mechanical water strider whose means of propulsion is analogous to that of its natural counterpart. PMID- 12904791 TI - Bottlenose dolphins perceive object features through echolocation. AB - How organisms (including people) recognize distant objects is a fundamental question. The correspondence between object characteristics (distal stimuli), like visual shape, and sensory characteristics (proximal stimuli), like retinal projection, is ambiguous. The view that sensory systems are 'designed' to 'pick up' ecologically useful information is vague about how such mechanisms might work. In echolocating dolphins, which are studied as models for object recognition sonar systems, the correspondence between echo characteristics and object characteristics is less clear. Many cognitive scientists assume that object characteristics are extracted from proximal stimuli, but evidence for this remains ambiguous. For example, a dolphin may store 'sound templates' in its brain and identify whole objects by listening for a particular sound. Alternatively, a dolphin's brain may contain algorithms, derived through natural endowments or experience or both, which allow it to identify object characteristics based on sounds. The standard method used to address this question in many species is indirect and has led to equivocal results with dolphins. Here we outline an appropriate method and test it to show that dolphins extract object characteristics directly from echoes. PMID- 12904792 TI - Neuronal populations and single cells representing learned auditory objects. AB - The neural representations associated with learned auditory behaviours, such as recognizing individuals based on their vocalizations, are not well described. Higher vertebrates learn to recognize complex conspecific vocalizations that comprise sequences of easily identified, naturally occurring auditory objects, which should facilitate the analysis of higher auditory pathways. Here we describe the first example of neurons selective for learned conspecific vocalizations in adult animals--in starlings that have been trained operantly to recognize conspecific songs. The neuronal population is found in a non-primary forebrain auditory region, exhibits increased responses to the set of learned songs compared with novel songs, and shows differential responses to categories of learned songs based on recognition training contingencies. Within the population, many cells respond highly selectively to a subset of specific motifs (acoustic objects) present only in the learned songs. Such neuronal selectivity may contribute to song-recognition behaviour, which in starlings is sensitive to motif identity. In this system, both top-down and bottom-up processes may modify the tuning properties of neurons during recognition learning, giving rise to plastic representations of behaviourally meaningful auditory objects. PMID- 12904793 TI - Neural correlates of implied motion. AB - Current views of the visual system assume that the primate brain analyses form and motion along largely independent pathways; they provide no insight into why form is sometimes interpreted as motion. In a series of psychophysical and electrophysiological experiments in humans and macaques, here we show that some form information is processed in the prototypical motion areas of the superior temporal sulcus (STS). First, we show that STS cells respond to dynamic Glass patterns, which contain no coherent motion but suggest a path of motion. Second, we show that when motion signals conflict with form signals suggesting a different path of motion, both humans and monkeys perceive motion in a compromised direction. This compromise also has a correlate in the responses of STS cells, which alter their direction preferences in the presence of conflicting implied motion information. We conclude that cells in the prototypical motion areas in the dorsal visual cortex process form that implies motion. Estimating motion by combining motion cues with form cues may be a strategy to deal with the complexities of motion perception in our natural environment. PMID- 12904794 TI - Induction of dendritic spines by an extracellular domain of AMPA receptor subunit GluR2. AB - Synaptic transmission from excitatory nerve cells in the mammalian brain is largely mediated by AMPA (alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid)-type glutamate receptors located at the surface of dendritic spines. The abundance of postsynaptic AMPA receptors correlates with the size of the synapse and the dimensions of the dendritic spine head. Moreover, long-term potentiation is associated with the formation of dendritic spines as well as synaptic delivery of AMPA receptors. The molecular mechanisms that coordinate AMPA receptor delivery and spine morphogenesis are unknown. Here we show that overexpression of the glutamate receptor 2 (GluR2) subunit of AMPA receptors increases spine size and density in hippocampal neurons, and more remarkably, induces spine formation in GABA-releasing interneurons that normally lack spines. The extracellular N terminal domain (NTD) of GluR2 is responsible for this effect, and heterologous fusion proteins of the NTD of GluR2 inhibit spine morphogenesis. We propose that the NTD of GluR2 functions at the cell surface as part of a receptor-ligand interaction that is important for spine growth and/or stability. PMID- 12904795 TI - Accelerated vaccination for Ebola virus haemorrhagic fever in non-human primates. AB - Containment of highly lethal Ebola virus outbreaks poses a serious public health challenge. Although an experimental vaccine has successfully protected non-human primates against disease, more than six months was required to complete the immunizations, making it impractical to limit an acute epidemic. Here, we report the development of accelerated vaccination against Ebola virus in non-human primates. The antibody response to immunization with an adenoviral (ADV) vector encoding the Ebola glycoprotein (GP) was induced more rapidly than with DNA priming and ADV boosting, but it was of lower magnitude. To determine whether this earlier immune response could nonetheless protect against disease, cynomolgus macaques were challenged with Ebola virus after vaccination with ADV GP and nucleoprotein (NP) vectors. Protection was highly effective and correlated with the generation of Ebola-specific CD8(+) T-cell and antibody responses. Even when animals were immunized once with ADV-GP/NP and challenged 28 days later, they remained resistant to challenge with either low or high doses of virus. This accelerated vaccine provides an intervention that may help to limit the epidemic spread of Ebola, and is applicable to other viruses. PMID- 12904797 TI - Physical paradoxes. PMID- 12904798 TI - First for physics Chicago. PMID- 12904796 TI - Crystal structure of the transfer-RNA domain of transfer-messenger RNA in complex with SmpB. AB - Accurate translation of genetic information into protein sequence depends on complete messenger RNA molecules. Truncated mRNAs cause synthesis of defective proteins, and arrest ribosomes at the end of their incomplete message. In bacteria, a hybrid RNA molecule that combines the functions of both transfer and messenger RNAs (called tmRNA) rescues stalled ribosomes, and targets aberrant, partially synthesized, proteins for proteolytic degradation. Here we report the 3.2-A-resolution structure of the tRNA-like domain of tmRNA (tmRNA(Delta)) in complex with small protein B (SmpB), a protein essential for biological functions of tmRNA. We find that the flexible RNA molecule adopts an open L-shaped conformation and SmpB binds to its elbow region, stabilizing the single-stranded D-loop in an extended conformation. The most striking feature of the structure of tmRNA(Delta) is a 90 degrees rotation of the TPsiC-arm around the helical axis. Owing to this unusual conformation, the SmpB-tmRNA(Delta) complex positioned into the A-site of the ribosome orients SmpB towards the small ribosomal subunit, and directs tmRNA towards the elongation-factor binding region of the ribosome. On the basis of this structure, we propose a model for the binding of tmRNA on the ribosome. PMID- 12904800 TI - Spontaneous tumescence after implantation of three-piece hydraulic penile prostheses: a short-term experience. AB - We evaluated spontaneous tumescence after penile prostheses implantation in a retrospective study. A total of 32 patients with three-piece hydraulic implants (17 Mentor Alpha I, 15 AMS Ultrex Plus) were enrolled. Consultation, personal interviews and questionnaires for both the patients and their partners provided the necessary information. The average follow-up period was 29 months. In total, 50 % of the patients reported spontaneous tumescence without activation of the implant and one further patient (3%) claimed full rigid spontaneous erections. General satisfaction with the prosthetic result was high at 91%. The following complications arose: one case of prosthesis infection, one case of mechanical failure, one case of cylinder dislocation and two cases of paraphimosis. A possible explanation for spontaneous tumescence after penile prosthesis implantation is the elevated preload of the compressed rather than destructed cavernous tissue. The artificial erection evoked by the prosthesis then takes the form of a normal physiological reaction and/or a good objective ridigity, and could account for the high patient satisfaction rate with three-piece hydraulic implants. PMID- 12904801 TI - Clomiphene increases free testosterone levels in men with both secondary hypogonadism and erectile dysfunction: who does and does not benefit? AB - Secondary hypogonadism is more common than primary gonadal failure and is seen in chronic and acute illnesses. Although testosterone has a role in erections, its importance in erectile dysfunction (ED) has been controversial. Hypogonadism produced by functional suppression of pituitary gonadotropins has been shown to correct with clomiphene citrate, but with a modest effect on sexual function. We wondered if longer treatment would produce improved results. A total of 178 men with secondary hypogonadism and ED received clomiphene citrate for 4 months. Sexual function improved in 75%, with no change in 25%, while significant increases in luteinizing hormone (P<0.001) and free testosterone (P<0.001) occurred in all patients. Multivariable analysis showed that responses decreased significantly with aging (P<0.05). Decreased responses also occurred in men with diabetes, hypertension, coronary artery disease, and multiple medication use. Since these conditions are more prevalent with aging, chronic disease may be a more important determinant of sexual dysfunction. Men with anxiety-related disorders responded better to normalization of testosterone. Assessment of androgen status should be accomplished in all men with ED. For those with lower than normal age-matched levels of testosterone treatment directed at normalizing testosterone with clomiphene citrate is a viable alternative to giving androgen supplements. PMID- 12904802 TI - Morphological modifications in clitoris and vagina in spontaneously hypertensive rats. AB - We evaluated possible morphological alteration in clitoris and vagina from spontaneous hypertensive rats (SHR) and normotensive WKY rats. Clitoris and vagina were processed by Masson's trichrome, anti-alpha-smooth-muscle actin, anticollagen type I (COL I) and type III (COL III), and anti-TGFbeta(1). SHR presented higher amount of clitoral cavernous smooth muscle (CSM), vascular smooth muscle; TGFbeta(1) in clitoral vessel wall; higher wall/lumen ratio in both vaginal and clitoral vessels; and remarkable interstitial fibrosis, expressed by a higher amount in interstitial COL I and III in both clitoris and vagina, compared to WKY rats. Nerve fibers from clitoral and vaginal tissue in SHR showed important fibrosis at perineurium. SHR showed positive correlation between systolic blood pressure (SBP) and clitoral CSM; SBP and fibrosis in clitoris; and SBP and COL I and III in clitoris, respectively. Similar findings were observed between SBP and COL I and III in vagina. In conclusion, SHR present morphologic changes in clitoral vessels as well as in clitoral cavernous space, which have a high positive correlation with the high blood pressure level. Moreover, the increase in extracellular matrix affects not only the clitoral and vaginal interstitium but also the nerve structures from both clitoris and vagina. PMID- 12904803 TI - Quality of Sexual Life Questionnaire (QVS): a reliable, sensitive and reproducible instrument to assess quality of life in subjects with erectile dysfunction. AB - A French quality of life questionnaire specific to erectile dysfunction (ED), 'QVS' for 'Questionnaire de Vie Sexuelle', has been developed. This paper describes its validation: item reduction and reliability (internal consistency and reproducibility), construct validity and criterion validity (clinical, discriminant and concurrent). The initial 40-item questionnaire was administered once to 316 ED and 117 control subjects, and twice (D0 and D7) to 104 ED and 29 control subjects. Item reduction gave a 27-item questionnaire with three scales (Sexual Life, Skills and Psychosocial Well-being) and four scores (one score for each scale and a Global Index). Psychometric analyses demonstrated the reliability and the validity of the QVS. The questionnaire was able to discriminate patients according to the presence or severity of ED. The Skills scale was the least sensitive. Analysis of responsiveness to change over time still needs to be addressed to consider the questionnaire as a fully validated instrument. PMID- 12904804 TI - Distribution of the Brief Male Sexual Inventory in community men. AB - The objectives of the study were to characterize male sexual functioning as related to age in community-dwelling older men. In 1989, a random sample of men aged 40-79 y (n=2115) without prior prostate surgery, prostate cancer, or other conditions known to affect voiding function (except benign prostatic hyperplasia) was invited (55% agreed) to participate in the Olmsted County Study of Urinary Symptoms and Health Status Among Men. In 1996, a previously validated male sexual function questionnaire was administered to the cohort. The questionnaire has 11 questions measuring sexual drive (two questions); erectile function (three) and ejaculatory function (two), as well as assessing problems with sex drive, erections, or ejaculation (three); and overall satisfaction with sex life (one). Each question is scored on a scale of 0-4, with higher scores indicating better functioning. Cross-sectional age-specific means (+/-s.d.) for drive, erections, ejaculation, problems, and overall satisfaction declined from 5.2 (+/-1.5), 9.8 (+/-2.5), 7.4 (+/-1.4), 10.7 (+/-2.2), and 2.6 (+/-1.0), respectively, for men in their 40s to 2.4 (+/-1.6), 3.3 (+/-3.4), 3.6 (+/-3.2), 7.7 (+/-3.8), and 2.1 (+/ 1.2) for men 70 y and older (all P<0.001). The cross-sectional decline in function with age was not constant, with age-related patterns differing by domain. The percentage of men reporting erections firm enough to have intercourse in the past 30 days declined from 97% (454/468) among those in their 40s to 51% (180/354) among those in their 80s (P&<0.001). In age-adjusted analyses, men reporting regular sexual partners had statistically significantly higher levels of sex drive, erectile function, ejaculatory function, and overall satisfaction than those who did not report regular sexual partners. Sexual drive, erectile functioning, ejaculatory functioning, and overall sexual satisfaction in men show somewhat differing cross-sectional patterns of decline with advancing age. Active sexual functioning is maintained well into the 80s in a substantial minority of community-dwelling men. PMID- 12904805 TI - Validation of the German version of the International Index of Erectile Function (IIEF) in patients with erectile dysfunction, Peyronie's disease and controls. AB - The objective of this study was to validate the German translation of the International Index of Erectile Function (IIEF). The IIEF was administered to 59 patients with erectile dysfunction (ED), to 38 patients with Peyronie's disease and to 33 controls. All patients were investigated by standardized German versions of international questionnaires of anxiety, depression, social desirability, quality of partnership, physical complaints and life-satisfaction. The five subscales of the English version, however, could not be replicated. Internal consistency for the complete questionnaire of 15 items was high (Cronbach's alpha=0.95). Based on the total scale and two subscales, we were able to discriminate sexual function between the ED group and the comparison groups. There was no significant correlation between the IIEF scores and anxiety, depression, somatic complaints and life-satisfaction. The German version of the IIEF has found good comprehension, and acceptance by the majority of patients. Its use is somewhat limited by its focus on sexual activity in partnership. In contrast to the English version, it mainly addresses a single factor of sexual function. PMID- 12904806 TI - Does sexual dysfunction correlate with deterioration of somatic sensory system in diabetic women? AB - To evaluate genital and extragenital somatic sensory system in diabetic women using biothesiometry and investigate the relation with sexual dysfunction. A total of 30 diabetic women and 20 normal sexually active women as a control group were evaluated with a detailed medical and sexual history including Index of Female Sexual Function (IFSF) questionnaire. Somatic sensory system of all women enrolled to the study was assessed by biothesiometry and threshold sensory values of nine genital sites and 14 extragenital sites were analyzed. The IFSF score in diabetic women was 23.6 while it was 38.3 in the control group (&<0.0005). For each genital as well as extragenital sites, the mean biothesiometric values were significantly higher in diabetics. The sensation of introitus vagina, labium minora and clitoris were found to be the most deteriorated genital sites in diabetic women. The difference between diabetic women with or without female sexual dysfunction (FSD) was not significant for biothesiometric values. Our data indicate that, somatic sensory system is affected by diabetes however sexual dysfunction does not always manifest. PMID- 12904807 TI - Apomorphine-induced brain modulation during sexual stimulation: a new look at central phenomena related to erectile dysfunction. AB - It is well recognized that sexual stimulation leading to penile erection is controlled by different areas in the brain. Animal erection studies have shown that apomorphine (a D2>D1 dopamine receptors nonselective agonist) seems to act on neurons located within the paraventricular nucleus and the medial preoptic area of the hypothalamus. Yet, only recently, was a centrally acting agent, apomorphine sublingual, approved for the treatment of erectile dysfunction. The present functional magnetic resonance imaging placebo-controlled study presents the first in vivo demonstration of the apomorphine-induced modulation of cortical and subcortical brain structures in patients with psychogenic erectile dysfunction. Noteworthy, patients in comparison with potent controls, showed an increased activity in frontal limbic areas that was downregulated by apomorphine. This suggests that psychogenic impotence may be associated with previously unrecognized underlying functional abnormalities of the brain. PMID- 12904810 TI - Sildenafil does not improve sexual function in men without erectile dysfunction but does reduce the postorgasmic refractory time. AB - Sildenafil is one of two oral drugs approved for first-line treatment of erectile dysfunction (ED). Anecdotally, some young healthy men who wish to enhance their sexual performance are requesting or abusing sildenafil. In this randomized double-blind, placebo-controlled clinical study, we investigated the effect of sildenafil in young men without ED. A total of 60 young healthy men age 20-40 y with no reported ED were enrolled for this single-dose home-use study. Subjects had used no medication in the 6 months prior to the study. All had been engaged in a stable relationship for at least 3 months. After completing the IIEF-5 questionnaire, patients were randomized in a double-blind fashion to receive either one 25 mg tablet of sildenafil (group 1) taken prior to intercourse, or an identical placebo tablet (group 2). All subjects completed a questionnaire relating to their erectile quality. There were no differences between the two groups in the reported improvement of erection quality, 12/30 sildenafil vs 10/30 placebo (Fisher's test, P=0.79). Sildenafil caused a significant reduction of the postejaculatory refractory time (12/30 vs 4/30) (chi(2) test, P=0.04). Sildenafil does not improve erections in young healthy men. Sildenafil should not be given to young healthy men to improve their erections and patients should be advised against recreational abuse of the drug. In this limited single-dose home study, sildenafil appears to reduce the postorgasmic refractory time. Although controlled studies are needed to evaluate the efficacy of erection-enhancing drugs in premature ejaculation, it is possible that sildenafil might be useful for this indication. PMID- 12904809 TI - Current drug use as risk factor for erectile dysfunction: results from an Italian epidemiological study. AB - Several drugs have been associated with an increased risk of erectile dysfunction (ED). We analysed the role of pharmacological treatments on the risk of ED using data from a cross-sectional study on prevalence and risk factors for ED in the general population in Italy. A total of 2450 men aged more than 18 years were randomly identified by 143 general practioners (GP) among their registered patients and invited to a confidential interview by their GP. Patients were asked 'about their ability to achieve and maintain an erection sufficient for satisfactory sexual performance'. If they were dissatisfied, they were defined as having ED. Out of the 2450 men identified, 440 (18%) refused to participate. The present analysis therefore includes information on 2010 men. After adjustment for related pathologies, anxiolytics and antidepressants showed insignificantly higher odds ratio (ORs, respectively, 1.7 and 2.1); antipsychotic drug use significantly increased the risk of ED (OR 9.0, 95% confidence interval, CI 1.8 44.4). Diuretics (OR 3.1, 95% CI 1.4-6.9) and anticholinergic drugs (OR 12.8, 95% CI 2.7-60.1) were associated with ED risk. No association emerged between ED and H2 antagonists, anticholesterolemic or hypoglycemic drugs. In conclusion, after taking account of related pathologies, our results suggest that men treated with antipsychotic, diuretic and anticholinergic drugs are at greater risk of ED. PMID- 12904808 TI - Structured interview on erectile dysfunction (SIEDY): a new, multidimensional instrument for quantification of pathogenetic issues on erectile dysfunction. AB - The aim of the present study is the definition of a brief structured interview (SI) providing scores useful for identification and quantification of pathogenetic factors of erectile dysfunction (ED). A SI was developed and applied to a consecutive series of 320 ED patients. A 13-item SI, with three-factor analysis-derived scales, was identified and applied for validation to an independent consecutive series of 194 ED patients. PGE(1) (10 microg) intracavernosal injection, penile duplex ultrasound (PDU), blood hormones, PSA, glycemia, and lipids were used for the assessment of an organic component (OC), and Middlesex Hospital Questionnaire (MHQ) modified for psychological disturbances. Scale 1, dealing with OC, showed a positive correlation with age, BMI, blood pressure, glycemia, and inverse correlation, with testosterone, PGE(1) and several parameters derived from PDU. Scale 2, related to partner's relationship, was not correlated with organic parameters. Scale 3, which measures psychopathological traits was correlated with MHQ scales. Scale 1 (>3) had a sensitivity of 67.9% and a specificity of 67.6% for OC. SIEDY((c)) provides information on ED pathogenesis and might assist physicians in diagnostic and therapeutic choices. PMID- 12904812 TI - Toxicity of antiretroviral therapy and implications for drug development. PMID- 12904811 TI - Pharmacophylogenomics: genes, evolution and drug targets. PMID- 12904813 TI - Therapeutic approaches to innate immunity: severe sepsis and septic shock. PMID- 12904814 TI - Mild cognitive impairment clinical trials. PMID- 12904815 TI - Logistics of process R&D: transforming laboratory methods to manufacturing scale. PMID- 12904816 TI - Drug research: myths, hype and reality. PMID- 12904817 TI - Opinion: understanding 'global' systems biology: metabonomics and the continuum of metabolism. PMID- 12904818 TI - Determining the position of the cell division plane. AB - Proper positioning of the cell division plane during mitosis is essential for determining the size and position of the two daughter cells--a critical step during development and cell differentiation. A bipolar microtubule array has been proposed to be a minimum requirement for furrow positioning in mammalian cells, with furrows forming at the site of microtubule plus-end overlap between the spindle poles. Observations in other species have suggested, however, that this may not be true. Here we show, by inducing mammalian tissue cells with monopolar spindles to enter anaphase, that furrow formation in cultured mammalian cells does not require a bipolar spindle. Unexpectedly, cytokinesis occurs at high frequency in monopolar cells. Division always occurs at a cortical position distal to the chromosomes. Analysis of microtubules during cytokinesis in cells with monopolar and bipolar spindles shows that a subpopulation of stable microtubules extends past chromosomes and binds to the cell cortex at the site of furrow formation. Our data are consistent with a model in which chromosomes supply microtubules with factors that promote microtubule stability and furrowing. PMID- 12904820 TI - [Not Available]. PMID- 12904819 TI - Replication of a cis-syn thymine dimer at atomic resolution. AB - Ultraviolet light damages DNA by catalysing covalent bond formation between adjacent pyrimidines, generating cis-syn cyclobutane pyrimidine dimers (CPDs) as the most common lesion. CPDs block DNA replication by high-fidelity DNA polymerases, but they can be efficiently bypassed by the Y-family DNA polymerase pol eta. Mutations in POLH encoding pol eta are implicated in nearly 20% of xeroderma pigmentosum, a human disease characterized by extreme sensitivity to sunlight and predisposition to skin cancer. Here we have determined two crystal structures of Dpo4, an archaeal pol eta homologue, complexed with CPD-containing DNA, where the 3' and 5' thymine of the CPD separately serves as a templating base. The 3' thymine of the CPD forms a Watson-Crick base pair with the incoming dideoxyATP, but the 5' thymine forms a Hoogsteen base pair with the dideoxyATP in syn conformation. Dpo4 retains a similar tertiary structure, but each unusual DNA structure is individually fitted into the active site for catalysis. A model of the pol eta-CPD complex built from the crystal structures of Saccharomyces cerevisiae apo-pol eta and the Dpo4-CPD complex suggests unique features that allow pol eta to efficiently bypass CPDs. PMID- 12904822 TI - [The importance of monitoring legionellosis in hospital setting: the case of a Sicilian Hospital]. AB - Hospital-acquired legionellosis is a major problem. In Italy this issue is still underestimated because the reported figures are lower than the actual cases. This is probably due to a lack of diagnosis and monitoring. In Sicily, no cases of hospital-acquired legionellosis were reported in the last few years. In the year 2001, a reference laboratory was set up to help health authorities monitor and prevent hospital-acquired legionellosis. The authors report the data concerning a measure of environmental monitoring and water treatment carried out in a hospital where no legionellosis was ever detected before. PMID- 12904823 TI - [Health and demographic status of unauthorized immigrants: the Albanian refugees landed in Italy (Apulia) in 1997]. AB - This study is aimed at assessing the health care demand of the Albanian groups arrived in Italy (Apulia) in 1997 and accommodated in the Cassano and Palese refugee camps. The analysis is based on the forms filled in on their arrival and first aid registers. Albanian health and demographic official data were also analyzed. The most common health problems observed amongst refugee groups were slight illnesses (23.3%), respiratory tract infections (11.3%), gastroenteritis (9.9%). In the light of these data, the refugees' status does not appear to be critical, although there remains the need to carry out constant monitoring to prevent infectious diseases and improve the quality of social facilities. PMID- 12904824 TI - [Microbiological quality of seaside sands: a beach in Latium]. AB - This study is focused on the microbiological quality of a sandy beach in the coastal area around Rome, Italy. The microbiological surveys were carried out on the sands collected both on the beach and on the waterline. A low-concentration of faecal bacteria (streptococci outnumbered Escherichia coli) and a constant rate of staphylococci were detected over the sampling period. Significant statistical correlations were calculated between yeasts and moulds, Escherichia coli and streptococci, streptococci and sulfite-reducing clostridium spores. This survey's data could be a baseline for future studies. PMID- 12904821 TI - [Assessment of a mass screening program for breast cancer in the Milan Health Unit 1 (ASL Provincia di Milano 1)]. AB - This study examines the early data of a breast cancer screening campaign (Progetto Donna) set up by the Milan Health Unit 1 in 1999. The authors have focused on some basic organizational aspects of such campaign and the patients' high response to the project showed a good efficiency of this initiative, aimed at informing and awakening people. The high rate of suspiciously positive results checked in several Hospitals, led to a careful assessment of the quality and the standards of radiology services. PMID- 12904825 TI - [Evolution of microbial flora and organic matter in bottled mineral water]. AB - A 15-week survey was carried out by the authors in order to detect the total microbial count and the quantity of organic matter in mineral waters bottled in dark glass containers and stored at room temperature. Two different and significant increases were registered at the 2nd and 11th week respectively. A relation between microbial count and organic matter was observed during the period examined. PMID- 12904826 TI - [Health director and clinical governance]. AB - Measures are needed to enhance health care quality. These measures should be implemented within the field of Clinical Governance. The authors have pointed out the skills required to the person in charge of Clinical Governance as well as the new role that the Health Director should play in case he intends to be in charge of Clinical Governance himself. PMID- 12904827 TI - [Etiology of rare diseases caused by novel viral agents: a review of the most recent literature]. AB - The authors describe the viral pathologies that have become rare in Italy and in the rest of the western world; they also analyze some novel agents mainly imported from the tropical and Asian Countries. For some of the microorganisms, they describe the various zones at risk, ways of transmission, clinical case, recent lab methods and preventive measures. PMID- 12904828 TI - [Bioethical guidelines for equity in public health. (National Committee for Bioethics. Rome, 25th May 2001)]. PMID- 12904829 TI - Characterisation of a novel class of polyamine-based neuroprotective compounds. AB - Prolonged cerebral ischaemia initiates complex intra- and inter-cellular signalling cascades ultimately resulting in neuronal death. Well-characterised mediators of ischaemic cell death are glutamate, free radicals and nitric oxide. Many drugs that block these mechanisms are neuroprotective in vitro, but have unfavourable side-effect profiles in man. We have recently demonstrated that the compound L-arginyl-3,4-spermidine (L-Arg3,4) is neuroprotective in vitro through an interaction with several of these mechanisms, and prevents ischaemic neurodegeneration in vivo with no gross side effects. In this study, we have used solid-phase combinatorial chemistry, to synthesise a number of analogues of L Arg3,4, and investigate the structure-activity relationship using an in vitro, organotypic hippocampal slice culture model of cerebral ischaemia. A number of molecular features were identified which were essential for the neuroprotective activity including the requirement for a positive charge and an amino acid in the L-configuration. Relatively minor alterations to both the terminal arginine and polyamine moieties significantly attenuated neuroprotective efficacy. Our data implies that these compounds are neuroprotective through a currently undefined mechanism rather than non-specific ionic interactions described previously for other polyamine-containing compounds. PMID- 12904831 TI - Genetic analysis of phosphomannomutase/phosphoglucomutase from Vibrio furnissii and characterization of its role in virulence. AB - The pmm gene from Vibrio furnissii, which encodes phosphomannomutase (PMM), was cloned and sequenced. The open reading frame consisted of 1,434 bp, encoding a polypeptide of 477 amino acids with a molecular mass of 53,325 Da. The predicted amino acid sequence of V. furnissii PMM showed high similarity with PMMs from other enteric bacteria, such as V. cholerae, Salmonella sp. and Escherichia coli. The PMM protein was overexpressed in E. coli as a His(6)-tagged recombinant protein. The estimated apparent K(m )and k(cat) values of the purified recombinant protein for mannose 1-phosphate were about 60 microM and 800 min(-1), respectively. To investigate the biochemical functions and the role of pmm in the virulence of V. furnissii, a pmm knock-out mutant was constructed by homologous recombination mutation. Under the various physical conditions, cell numbers of the wild-type and the mutant did not differ. Oral introduction of bacterial suspensions to a mouse model showed that the pmm-deficient mutant decreased in viability at the intestine. Microscopy of the isolated intestines from mice revealed significant damage after 3 days in intestinal mucosa infected with the wild-type as compared with the mutant. The pmm-deficient mutant caused a reduction of virulence in mice and the loss of O-antigen polysaccharide, and showed low resistance relative to the wild-type when incubated with normal human serum. PMID- 12904832 TI - Fructose-1,6-bisphosphatase from Corynebacterium glutamicum: expression and deletion of the fbp gene and biochemical characterization of the enzyme. AB - The class II fructose-1,6-bisphosphatase gene of Corynebacterium glutamicum, fbp, was cloned and expressed with a N-terminal His-tag in Escherichia coli. Purified, His-tagged fructose-1,6-bisphosphatase from C. glutamicum was shown to be tetrameric, with a molecular mass of about 140 kDa for the homotetramer. The enzyme displayed Michaelis-Menten kinetics for the substrate fructose 1,6 bisphosphate with a K(m) value of about 14 micro M and a V(max) of about 5.4 micro mol min(-1) mg(-1) and k(cat )of about 3.2 s(-1). Fructose-1,6 bisphosphatase activity was dependent on the divalent cations Mg(2+) or Mn(2+) and was inhibited by the monovalent cation Li(+) with an inhibition constant of 140 micro M. Fructose 6-phosphate, glycerol 3-phosphate, ribulose 1,5 bisphosphate and myo-inositol-monophosphate were not significant substrates of fructose-1,6-bisphosphatase from C. glutamicum. The enzymatic activity was inhibited by AMP and phosphoenolpyruvate and to a lesser extent by phosphate, fructose 6-phosphate, fructose 2,6-bisphosphate, and UDP. Fructose-1,6 bisphosphatase activities and protein levels varied little with respect to the carbon source. Deletion of the chromosomal fbp gene led to the absence of any detectable fructose-1,6-bisphosphatase activity in crude extracts of C. glutamicum WTDelta fbp and to an inability of this strain to grow on the carbon sources acetate, citrate, glutamate, and lactate. Thus, fbp is essential for growth on gluconeogenic carbon sources and likely codes for the only fructose-1,6 bisphosphatase in C. glutamicum. PMID- 12904830 TI - Therapeutic administration of 3,4,5-trimethoxy-4'-fluorochalcone, a selective inhibitor of iNOS expression, attenuates the development of adjuvant-induced arthritis in rats. AB - We have previously investigated the effects of a series of dimethoxy- and trimethoxychalcone derivatives, with various patterns of fluorination, on nitric oxide production in LPS-stimulated murine RAW 264.7. The present study was designed to determine if 3,4,5-trimethoxy-4'-fluorochalcone (CH 17) could modulate the production of NO and/or prostaglandins in vivo. On the mouse macrophage cell line RAW 264.7 CH 17 inhibited dose-dependently NO production, with an IC(50) value in the nanomolar range, and reduced PGE(2) levels by a 58% at 10 microM. This compound had no direct inhibitory effect on iNOS and COX-2 activities. NO reduction was the consequence of inhibition of the expression of iNOS. In vitro experiments indicated that CH 17 is an inhibitor of the nuclear factor-kappaB (NF-kappaB) pathway of cellular activation in macrophages. This compound exhibited in vivo an inhibitory behaviour correlated with its in vitro results on nitrite and PGE(2) accumulation. In the rat adjuvant-induced arthritis, oral administration of CH 17 (25 mg/kg) on days 17-24 after adjuvant injection, significantly inhibited paw oedema, protected from weight loss and reduced the levels of inflammatory mediators (nitrites and PGE(2)) in paw homogenates, without affecting PGE(2) levels in stomach homogenates. The profile and potency of this compound, a selective inhibitor of iNOS expression that interferes with NF-kappaB activation, may have relevance for the inhibition of the inflammatory response, representing a new approach to the modulation of different inflammatory pathologies. PMID- 12904833 TI - Mortality and morbidity associated with osteoporosis drug treatment following hip fracture. AB - This study examined post-fracture osteoporosis drug treatment in hip fracture patients and the association of treatment with mortality and morbidity. Pre- and post-fracture demographic/health information was collected on a cohort of hip fracture patients aged 65+ years. Post-fracture administrative data on prescription drug use and health care utilization was linked to the cohort data. Five classes of osteoporosis drugs were available during the study period: hormone replacement therapy (HRT), bisphosphonates (BSP), calcitonin, selective estrogen receptor modulators (SERMs) and vitamin D(3) (Rocaltrol). Pre-fracture, 38 of 449 patients (8%) were on osteoporosis medications. Post-fracture, 81 of 356 patients (23%) were treated; 63 of these patients were untreated prior to fracture. Both treated and untreated patients had similar rates of subsequent hip fracture (6% and 4%, respectively) and Colles fracture (2%). Regardless of treatment status, patients were also equally likely to be hospitalized, both in the short-term (28% in treated, 27% in untreated) and in the long-term (43% versus 37%). However, mortality was significantly lower in the treated group. The lower mortality in the treated group, combined with the knowledge that antiresorptive drugs reduce fractures and increase bone density, merit undertaking a randomized trial to confirm our findings that antiresorptive therapy should be considered in all patients post-hip fracture. PMID- 12904834 TI - Critical appraisal of physical rehabilitation measures after osteoporotic vertebral fracture. AB - Osteoporosis is asymptomatic until fracture occurs. Fracture of the vertebral bodies can be "silent" and is diagnosed incidentally on radiographic visualization. The occurrence of spontaneous vertebral fracture in an otherwise healthy individual is considered pathognomonic of spinal osteoporosis. Osteoporotic vertebral fractures and kyphotic posture are no longer disorders about which nothing can be done. Resistive training exercises can decrease the risk of vertebral fractures. Proprioception generated within joints, ligaments, and muscles contributes to awareness of the relative orientation of functional units of the spine at rest and in motion. This is fundamental to posture, balance, and locomotion. Proprioception reeducation can be utilized for improving posture and balance. The focus of this review is rehabilitative measures for management of vertebral fractures. PMID- 12904835 TI - Osteoporotic fracture: missed opportunity for intervention. AB - As prior fracture is consistently associated with increased risk of subsequent fracture, subjects with a history of prior fracture represent a high risk group which should be targeted for intervention to reduce future fracture rates. The aim of this study was to investigate whether prior osteoporotic fracture affected treatment patterns among subjects admitted with hip fractures. All hip fracture admissions to two major teaching hospitals of the University of New South Wales, Sydney, Australia, over the 12-month period between July 1997 and June 1998 were identified retrospectively from medical records. Patient demographics, frequency and location of prior fractures, and treatment status on admission were recorded. There were a total of 348 atraumatic hip fracture admissions over this 12-month period. Forty five percent of 251 women and 30% of 97 men with an osteoporotic hip fracture had a known prior fracture, including prior hip fracture in 19% of the women and 8% of the men. Among subjects with prior fractures, only 18% of women and 7% of men were on any specific anti-osteoporosis therapy. Even among those with a prior hip fracture, only 21% of women and none of the men were taking optimal appropriate therapy. A high proportion of individuals suffering hip fractures had sustained prior "signal" fractures. Although more subjects with prior fracture received treatment than those without prior fracture, total treatment levels were low, and the majority of high-risk subjects did not receive therapy shown to reduce the risk of further fractures. PMID- 12904836 TI - Quantitative ultrasound assessment of acute bone loss following spinal cord injury: a longitudinal pilot study-comment. PMID- 12904837 TI - A theoretical analysis of the relative influences of peak BMD, age-related bone loss and menopause on the development of osteoporosis. AB - Factors that determine a post-menopausal woman's bone mineral density (BMD) include her mass at the time of skeletal maturity (peak BMD), menopause and the rate of loss she experiences as she ages. Understanding the relative influence of each of these factors may help identify important preventive treatments and provide new ways to identify women at risk for osteoporosis. In this analysis we utilize a computer model of the bone remodeling process to predict the relative influences of peak BMD, menopause and age-related bone loss on the development of osteoporosis. The delay in the onset of osteoporosis (defined as BMD <2.5 SD from the young adult mean) caused by modifying peak BMD, age-related bone loss or the age at menopause is quantified. A 10% increase in peak BMD is predicted to delay the development of osteoporosis by 13 years, while a 10% change in the age at menopause or the rate of non-menopausal bone loss is predicted to delay osteoporosis by approximately 2 years, suggesting that peak BMD may be the single most important factor in the development of osteoporosis. PMID- 12904839 TI - Effect of sub-elite competitive running on bone density, body composition and sexual maturity of adolescent females. AB - The attainment of optimal peak bone mass during adolescence is important in the primary prevention of osteoporosis. Exercise may contribute to skeletal development and bone density during growth, although competitive exercise is suggested to have an adverse effect. This study assesses the effect of moderate exercise on the bone density of adolescent females. Additionally, other factors which significantly influence attainment of peak bone mass were identified. This was a cross-sectional study of 42 adolescent females, classified as runners (n=15) or non-runners (n=27). Nutrient intake, energy expenditure, menstrual history and pubertal stage were recorded. Bone age, skinfold thickness, body composition and bone mineral density (BMD) of total body, lumbar spine and proximal femur were measured. Statistical analyses used Student's t-test, Pearson correlation and multiple regression analyses. Runners had lower fat mass and higher lean mass, with a trend to higher BMD in all sites measured. There were no significant differences in menstrual cycle regularity, age at menarche or number who had attained menarche. Pubic hair development was similar in both groups. Breast development was delayed in runners, although this may have been a function of lower fat mass in this group. When subjects were categorized according to menarchal status, postmenarchal girls were significantly taller and heavier, with higher fat mass and significantly higher total body and lumbar spine BMD. There was no significant relationship between BMD at any site and dietary nutrient intake. Multiple regression analyses, using BMD as the dependent variable, identified running status, pubertal stage, fat mass and lean mass as significant determinants. When BMD/height was used, significant determinants in total body BMD were fat mass, pubertal stage and running status, while in the lumbar spine, only the latter two variables remained significant. In conclusion, body composition, physical activity and sexual maturity were identified as significant determinants of bone density during adolescence. Runners had significantly lower body fat than non-runners, but this did not interfere with hormonal cyclicity, and caused no detriment to their bone density. The results of this study are reassuring, since they indicate that sub-elite competitive athletics has no detrimental effect on bone mass accrual in adolescent females. PMID- 12904840 TI - Effects of dietary improvement on bone metabolism in elderly underweight women with osteoporosis: a randomised controlled trial. AB - Malnutrition in elderly people contributes to osteoporosis and fracture. The aim of the study was to investigate the effects of nutritional improvement on bone metabolism in elderly community-dwelling women. A 12-month randomized controlled trial of 71 ambulant women aged > or =70 years with BMI < or =21 kg/m(2 )and osteoporosis at the hip was undertaken. They received either calcium (1 g) and vitamin D (800 units of cholecalciferol) only (group 1: n=35) or calcium/vitamin D and one or two cartons of a nutritional supplement drink which provided 300 Kcal, 12 g protein, 11.6 g fat and 36.8 g carbohydrate per carton (group 2: n=36). Body composition and bone mineral density (BMD) were assessed at baseline and 12 months. Biochemical markers of bone turnover were measured at baseline and at 1, 3, 6, 9 and 12 months. Group 2 gained significantly more weight [mean (SD) group 1: 0.15 (2.45), group 2:2.66 (2.8) kg P<0.001] and fat mass [group 1: -0.26 (1.8), group 2:1.9 (1.7) kg P<0.001]. BMD at the spine, femoral neck and total hip did not change significantly, although there was a positive trend at the total hip in group 2 [group 1: -0.5 (5.2), group 2:1.25 (3.3)%, P=0.13]. In a subgroup analysis, irrespective of their treatment group, there was a significant difference in changes in BMD at the lumbar spine and total hip in those who lost body weight (A) compared to those who had maintained or increased their weight (B), [mean (SD) % change in BMD lumbar spine; A: -1.64 (3.75), B: 0.96 (2.75) P=0.013, total hip A: -2.09 (6.0), B: 1.04 (3.3), P=0.05)] A significant reduction in serum CTX, a marker of bone resorption, was seen in group 2 [% decrease at 3 months, group 1: 1 (8.7), Group 2: 32 (5.8), P<0.01]. Serum osteoprotegerin (OPG) increased significantly in group 2 with a maximal increase (27%) observed at 6 ( P<0.01) and 9 months ( P<0.05). A small increase in bone specific alkaline phosphatase was seen at 12 months in group 2 [% increase group 1:5 (5), group 2: 17 (6), P=0.05]. Serum osteocalcin increased at 12 months in group 2 ( P=0.01). Dietary improvement in elderly women with low BMI is associated with a reduction in bone resorption with a small but "net" positive effect on bone formation. PMID- 12904841 TI - Influence of muscle strength, physical activity and weight on bone mass in a population-based sample of 1004 elderly women. AB - High physical activity level has been associated with high bone mass and low fracture risk and is therefore recommended to reduce fractures in old age. The aim of this study was to estimate the effect of potentially modifiable variables, such as physical activity, muscle strength, muscle mass and weight, on bone mass in elderly women. The influence of isometric thigh muscle strength, self estimated activity level, body composition and weight on bone mineral density (dual energy X-ray absorptiometry; DXA) in total body, hip and spine was investigated. Subjects were 1004 women, all 75 years old, taking part in the Malmo Osteoporosis Prospective Risk Assessment (OPRA) study. Physical activity and muscle strength accounted for 1-6% of the variability in bone mass, whereas weight, and its closely associated variables lean mass and fat mass, to a much greater extent explained the bone mass variability. We found current body weight to be the variable with the most substantial influence on the total variability in bone mass (15-32% depending on skeletal site) in a forward stepwise regression model. Our findings suggest that in elderly women, the major fracture-preventive effect of physical activity is unlikely to be mediated through increased bone mass. Retaining or even increasing body weight is likely to be beneficial to the skeleton, but an excess body weight increase may have negative effects on health. Nevertheless, training in elderly women may have advantages by improving balance, co-ordination and mobility and therefore decreasing the risk of fractures. PMID- 12904842 TI - Diagnosis of chondral lesions of the knee joint: can MRI replace arthroscopy? A prospective study. AB - The role of magnetic resonance tomography (MRI) for the diagnosis of chondral lesions of the knee joint is still unclear. The sensitivity of the method ranges from 15% to 96%. The scope of our daily experiences showed that there were considerable deviations between the tomographical and arthroscopical results, which vary from the results of experimental studies. Therefore we have conducted a prospective study to investigate the question of how MRI can replace arthroscopy (ASC) in the diagnosis of cartilage damages in the scope of daily routine. All 195 patients included in this study received a magnetic resonance tomography followed by an arthroscopy. A clear diagnosis of supposition had to be determined before the magnetic resonance tomography. The patients were divided into 3 Groups. Group A (n=86) received a standard Military Hospital Ulm (MH) MRI- sagittal STIR TSE and PD TSE, coronal and transversal T2 FFE (TR=660 ms, TE=18 ms, FA=30 degrees, 512 matrix). In addition, one sub-Group, AK (n=21) was examined with a special cartilage sequence of the cartilage fs T1 W FFE. Neither patients in Group AK nor in Group A as a whole received any contrast medium. Group B (n=88) was examined with an alternate MRI protocol (Radiological Joint Practice, Neu-Ulm--sagittal T1 SE, T2 SE and T2 FLASH (TR=608 ms, TE=18 ms, FA=20 degrees, 256 matrix), coronal PD fs), employing gadolinium as a contrast medium. 156 cartilage lesions were found arthroscopically. In Group A the sensitivity was 33%, the specificity 99%, and the positive and negative prediction values 75% and 98% respectively. Group B reached a sensitivity of 53% and a specificity of 98%. The positive prediction value was 48% and the negative was 98%. Group AK showed a sensitivity of 38% and specificity of 98%; the positive and negative prediction values came to 50% and 97% respectively. In conclusion, our results indicate that the MRI examination techniques recommended in the literature at present are not able to replace the ASC for the diagnosis of cartilage damages of the knee joint. In view of the high specificity (97%-99%) and the high negative prediction value (97%-98%), MRI is suitable for the exclusion of cartilage lesions. For a negative MRI associated with a cartilage injury, a cautious attitude towards an operative cartilage treatment is therefore justified. Because the MRI can not replace the ASC for diagnostic of cartilage damage, the ASC still has to be seen as the method of choice for the evaluation of cartilage damage. PMID- 12904843 TI - Glycogenosis storage type I diseases and evolutive adenomatosis: an indication for liver transplantation. AB - We report on two cases of type I glycogen storage disease (GSD) complicated by malignant tumors. A 23-year-old man had GSD Ia with adenomatosis. He underwent transplantation for rapidly growing and radiologically changing adenomata. At histological examination, one adenoma had become a hepatocellular carcinoma. A 22 year-old, HBV-infected woman had GSD type Ib with adenomatosis. At follow-up, several tumors showed changing morphological characteristics. Pre-transplant laparotomy confirmed the presence of a metastatic cholangiocarcinoma. Liver transplantation should be considered in GSD type I patients with adenomatosis, especially when tumor characteristics change. Regular detailed Doppler ultrasound and magnetic nuclear resonance screening during childhood and adolescence are, therefore, mandatory in order for the timing of transplantation to be optimized. PMID- 12904838 TI - Searching for osteoporosis genes in the post-genome era: progress and challenges. AB - Osteoporosis is a common skeletal disease characterized by low bone mineral density (BMD), deterioration of bone microarchitecture and increased fracture risk. It is a complex disease that has high social and economic costs. Osteoporosis and its associated phenotypes are under the strong genetic control. Identification and characterization of specific loci or genes involved in determining osteoporosis and its associated phenotypes will contribute to a greater understanding of the pathogenesis of osteoporosis, and ultimately might lead to the development of better diagnosis, prevention and treatment strategies. Efforts to identify osteoporosis genes have focused on three approaches: animal models, candidate gene approach, and genome-wide scans. In this article, we review the current status for mapping and identification of genes for osteoporosis, with a focus on some promising regions and future prospects. PMID- 12904844 TI - Low-dose immunosuppression in a rat hind-limb transplantation model. AB - Composite tissue allografts (CTAs) offer an alternative to conventional reconstructive methods. However, the toxicity of the drugs that are required to prevent rejection has prevented its widespread clinical application. The purpose of this study was to determine whether a low-dose, corticosteroid-free combination regimen of tacrolimus and mycophenolate mofetil (MMF) would prevent rejection in a rat hind-limb model, with minimal toxic side effects. Three groups were used in this study. In group I, Wistar Furth (WF) rats received a syngeneic WF hind-limb. In groups II and III, WF rats received an ACI hind-limb. The latter were treated with tacrolimus-MMF. Assessment for rejection, flow cytometry, and mixed lymphocyte reactions was performed. Biopsies were taken regularly and at the time of killing. Combination therapy with low-dose tacrolimus-MMF effectively prolonged CTA survival indefinitely, with minimal side effects. Toxicity associated with immunosuppressive drugs can be avoided in a low-dose combination corticosteroid-free regimen. PMID- 12904845 TI - Clinical relevance of antibodies to HLA antigens undetectable by the standard complement-dependent cytotoxicity test. AB - Recent literary data suggest that antibodies to HLA antigens undetectable by the standard complement-dependent cytotoxicity test may cause not only chronic, but also acute immunological complications after kidney transplantation. The aim of this study was to investigate the significance of non-cytotoxic antibodies to HLA antigens for the development of immunological complications and a worse graft prognosis after first kidney transplantation. Sera before and early after transplantation from 120 first kidney recipients were analyzed by flow cytometry (FCXM), ELISA and the standard complement-dependent cytotoxicity (CDC) test. Pre transplant FCXM negativity was related to a lower incidence of rejection episodes in the first post-transplant year ( P<0.01). A significant association between acute rejection and the presence of antibodies to HLA class II antigens before and after transplantation was also found ( P<0.05). Our study supports the findings of other centers of the detrimental role to the kidney graft played by anti-HLA antibodies undetectable by the classical CDC test. PMID- 12904846 TI - Late cytomegalovirus disease following liver transplantation. AB - The widespread use of antiviral prophylaxis or preemptive therapy among orthotopic liver transplantation (OLT) recipients has reduced the occurrence of early cytomegalovirus (CMV) disease. Late disease is increasingly reported. Little is known about CMV disease occurring beyond the first year after transplantation. The aim of this study was to evaluate the occurrence of CMV disease two or more years after OLT and to determine its risk factors and clinical features. Eighty-one consecutive OLT recipients followed for 2 years or longer after transplantation were included in the study. Data were collected on demographic and clinical variables, clinical presentation, treatment, and outcome of late CMV disease. Late CMV disease occurred in 7/81 liver recipients (8.5%) at a mean time of 5.9 years after OLT (range: 3.5--9.3, median: 6.3 years). All seven patients were women, with a mean age of 47.7 years (range: 26--60, median: 59 years). There was no association between the development of late CMV disease and the occurrence of rejection episodes, treatment with corticosteroids, or the early use of antiviral prophylaxis. Clinical presentation included fever and disturbed liver functions in all patients, one patient had concurrent CMV pneumonitis and one CMV retinitis. Though all patients responded to ganciclovir, two had recurrent disease episodes and one patient died of secondary bacterial sepsis. Late-onset CMV disease can occur several years after OLT. Although it manifests classic clinical features of early disease, it is not associated with traditional risk factors and its pathogenesis may differ from that of early disease. PMID- 12904847 TI - Difficult weaning from mechanical ventilation following Wernicke's syndrome developing after gastroplasty. PMID- 12904848 TI - Acute community-acquired bacterial meningitis in adults admitted to the intensive care unit: clinical manifestations, management and prognostic factors. AB - OBJECTIVE: To study the clinical features, management and prognostic factors associated with adverse clinical outcome in a series of patients with acute community-acquired bacterial meningitis admitted to the intensive care unit (ICU). DESIGN AND SETTING: Descriptive, prospective study at two ICUs of a university hospital over a 6-year period. PATIENTS: Sixty-four episodes in 62 adults with acute community-acquired bacterial meningitis admitted to the ICU. RESULTS: Most of the patients (95.3%) were admitted to the ICU presenting with altered mental status (the median value of Glasgow Coma Scale (GCS) was 11). Overall mortality rate was 10.9% (7 patients) and 11 (17.1%) developed adverse clinical outcome (death or severe neurologic deficit). The features associated with adverse clinical outcome were: age over 50 years, seizures or focal neurologic signs at admission, a GCS score of 10 or less and an APACHE II score more than 13 at admission to the ICU. Only the severity of the disease determined according to the APACHE II scale was independently associated with adverse clinical outcome after a multivariate analysis was performed (adjusted odds ratio =8.74; 95% CI =1.70-44.77; p=0.009). All patients were empirically treated with third-generation cephalosporins and dexamethasone was used in 40 cases (62.5%). Ten patients (15.6%) received mannitol, nine of them after a transcranial Doppler sonography recording had been performed. CONCLUSIONS: In adult patients with acute community-acquired bacterial meningitis admitted to the ICU, the overall severity of the disease within 24 h of admission may be the major indicator of adverse in-hospital clinical outcome. PMID- 12904849 TI - Life-threatening hemoptysis in adults with community-acquired pneumonia due to Panton-Valentine leukocidin-secreting Staphylococcus aureus. AB - Three new consecutive cases of life-threatening hemoptysis in adults with community-acquired pneumonia due to Panton-Valentine leukocidin-secreting Staphylococcus aureus are presented, focusing on the particular clinical presentation of this new entity. Between December 1999 and March 2001, three adults aged from 23 to 67 years were admitted to our respiratory intensive care unit for massive hemoptysis and septic shock associated with community-acquired Staphylococcus aureus pneumonia. Isolates were sent to the Centre National de Reference des Toxemies Staphylococciques in Lyon, France, where they were found to secrete Panton-Valentive leukocidin. The clinical course was similar in the three patients, with massive hemoptysis and septic shock necessitating mechanical ventilation. Two patients died rapidly; necropsy showed pulmonary vascular necrosis in one of them. The third patient recovered after appropriate antibiotic therapy. Leukocidin/neutrophil interactions in the pulmonary vasculature may cause severe hemoptysis in patients with community-acquired Staphylococcus aureus pneumonia secreting Panton-Valentine leukocidin. Adult patients with massive hemoptysis and suspected community-acquired pneumonia should receive antibiotic regimens covering Staphylococcus aureus. PMID- 12904850 TI - Middle cerebral artery flow velocity decreases and electroencephalogram (EEG) changes occur as acute hypercapnia reverses. AB - OBJECTIVE: To define the changes in middle cerebral artery flow velocity (Vmca) and the electroencephalogram (EEG) during rapid reduction in arterial carbon dioxide (PCO2) from acute hypercapnia. DESIGN. Human volunteer study. SETTING: University-affiliated hospital experimental laboratory. PARTICIPANTS: Nine healthy volunteers aged 25-35 years. INTERVENTIONS: Subjects rebreathed exhaled gas from a bag pre-filled with 5% carbon dioxide (CO2) in oxygen, up to an end tidal CO2 of 10% or to the limit of discomfort, when they were disconnected. MEASUREMENTS AND RESULTS: Middle cerebral artery blood flow velocity was continuously measured by transcranial Doppler ultrasound, quantitative EEG was recorded and hemodynamics were monitored non-invasively. Vmca closely correlated with end-tidal CO2 changes (r=0.65, p<0.001). When rebreathing ceased, there was a rapid decline in end-tidal CO2 and in Vmca to baseline within 42+/-14 s, followed by a rapid further decline in both variables to below baseline. End tidal CO2 reached a nadir of 4.4+/-1.1% at 146+/-79 s, with Vmca decreasing to 37+/-10 cm/s at 104+/-65 s (a 40% reduction and 31% below basal values). Electroencephalogram alpha activity significantly decreased and delta activity increased during hypercapnia. During subsequent hypocapnia, delta activity decreased back to baseline, while alpha activity increased, but remained below baseline. CONCLUSIONS: These findings demonstrate the rapid changes occurring in cerebral blood flow during rapid declines in arterial CO2 and the consequent potential for producing brain ischemia and hemodynamic alterations if inadvertent hyperventilation occurs following institution of mechanical ventilation for acute hypercapnia. PMID- 12904851 TI - The role of hyperbaric oxygen in the management of subarachnoid hemorrhage. AB - OBJECTIVE: To determine the role of hyperoxic and hyperbaric therapy following experimental subarachnoid hemorrhage (SAH). DESIGN: Prospective, randomized, controlled animal study. SUBJECTS: Thirty male Wistar rats. INTERVENTIONS: Thirty rats were assessed for an initial neurologic status as double-blinded by two different neurosurgeons using a neurologic severity score (NSS) and then underwent an initial angiographic examination. Two days later, 0.3 ml of homologous blood was injected into the cisterna magna to produce a SAH-induced cerebral vasospasm. The NSS and angiographic examination were then repeated. The rats having no spasm or a spasm under 50% (n=8) and 50% or over 50% (n=22) were grouped separately, as groups 1 and 2, respectively. The rats having 50% or more spasm were further divided randomly into group 2A and 2B. The rats in groups 1 and 2A (n=11) underwent a 60-min course of 100% oxygen at the atmospheric pressure 1 atmosphere absolute (ata), and group 2B (n=11) received 100% oxygen at 3 ata for 1 h. Neurologic assessment was repeated on the next day and 7 days later. MEASUREMENTS AND MAIN RESULTS: The animals having no spasm or less than 50% spasm had a better NSS and outcome when compared with the animals having 50% or more spasm. But the animals with 50% or more spasm which underwent hyperbaric therapy were shown to have a better outcome compared to the animals having hyperoxic therapy. CONCLUSION: Exposure to hyperbaric oxygen therapy seemed to accelerate the recovery of neurologic deficits secondary to experimental SAH. PMID- 12904853 TI - Posterior reversible encephalopathy syndrome (PRES) in critically ill obstetric patients. AB - OBJECTIVE: To describe clinical, neuroradiological and evolutionary findings in obstetric patients with posterior reversible encephalopathy syndrome (PRES). DESIGN: Retrospective case series. SETTING: University intensive care unit (ICU). PATIENTS: Four critically ill patients. Two patients experienced PRES in late postpartum without the classical pre-eclamptic signs. All patients showed impairment of consciousness and epileptic seizures; two of them presented cortical blindness and headache, too. True status epilepticus (SE) occurred in two cases. In all patients MRI showed the typical feature of gray-white matter edema, mainly localized to the temporo-parieto-occipital areas. INTERVENTIONS: Normalization of high blood pressure (BP) and treatment of seizures. Two patients with SE and severe impairment of consciousness were treated with an intravenous valproate (ivVPA) bolus followed by continuous infusion. MEASUREMENTS AND RESULTS: In three cases, neurological and MRI abnormalities completely resolved in about a week. Another patient died due to subarachnoid hemorrhage. CONCLUSION: Posterior reversible encephalopathy syndrome is a well described clinical and neuroradiological syndrome characterized by headache, altered mental status, cortical blindness and seizures, and a diagnostic MRI picture; usually reversible, PRES can sometimes result in death or in irreversible neurological deficits, thus requiring early diagnosis and prompt treatment. PRES can have various etiologies, but pregnancy and postpartum more frequently lead to this condition. Treatment of seizures deserves special attention since the anti epileptic drugs currently used in SE management may worsen vigilance as well as autonomic functions. Extensive research is needed to assess the role of ivVPA in this condition. PMID- 12904854 TI - Influence of tidal volume on stroke volume variation. Does it really matter? PMID- 12904852 TI - The pathophysiology of propofol infusion syndrome: a simple name for a complex syndrome. AB - Propofol infusion syndrome (PRIS) is a rare and often fatal syndrome described in critically ill children undergoing long-term propofol infusion at high doses. Recently several cases have been reported in adults, too. The main features of the syndrome consist of cardiac failure, rhabdomyolysis, severe metabolic acidosis and renal failure. To date 21 paediatric cases and 14 adult cases have been described. These latter were mostly patients with acute neurological illnesses or acute inflammatory diseases complicated by severe infections or even sepsis, and receiving catecholamines and/or steroids in addition to propofol. Central nervous system activation with production of catecholamines and glucocorticoids, and systemic inflammation with cytokine production are priming factors for cardiac and peripheral muscle dysfunction. High-dose propofol, but also supportive treatments with catecholamines and corticosteroids, act as triggering factors. At the subcellular level, propofol impairs free fatty acid utilisation and mitochondrial activity. Imbalance between energy demand and utilisation is a key pathogenetic mechanism, which may lead to cardiac and peripheral muscle necrosis. Propofol infusion syndrome is multifactorial, and propofol, particularly when combined with catecholamines and/or steroids, acts as a triggering factor. The syndrome can be lethal and we suggest caution when using prolonged (>48 h) propofol sedation at doses higher than 5 mg/kg per h, particularly in patients with acute neurological or inflammatory illnesses. In these cases, alternative sedative agents should be considered. If unsuitable, strict monitoring of signs of myocytolysis is advisable. PMID- 12904855 TI - Swallowing disorders post orotracheal intubation in the elderly. AB - OBJECTIVES: The purpose of this study was to assess the prevalence and recovery time of swallowing dysfunction after prolonged endotracheal intubation in critically ill elderly patients compared to a younger cohort. DESIGN: This was a prospective, interventional, clinical study set in a medical intensive care unit in a university-affiliated hospital. SUBJECTS: The study involved 42 consecutive elderly patients (>/=65 years old) and 42 controls (<65 years) matched for severity of illness requiring endotracheal intubation for more than 48 h. INTERVENTIONS: A fiberoptic endoscopic evaluation of swallowing (FEES) was performed within 48 h post-extubation and on days 5, 9, and 14 for those with evidence of aspiration. RESULTS. Swallowing dysfunction was assessed by the detection of test material below the true vocal cords. Aspiration was documented in 52% of the elderly and 36% of the control group (P=0.2). No significant difference in the co-morbidity index and the length of mechanical ventilation was found between aspirators and non-aspirators. None of the control group had swallowing deficits after 2 weeks, while 13% of the elderly participants showed persistent impairment in the swallowing reflex. By multivariate analysis, the preadmission functional status was the only determinant of a slowly resolving swallowing deficit (hazard ratio 1.68; 95% confidence interval 1.26-3.97). No post-extubation aspiration pneumonia was identified in either group. CONCLUSIONS: Critically ill elderly patients exhibit delayed resolution of swallowing impairment post extubation. FEES should be considered for those with impaired preadmission functional status. PMID- 12904856 TI - An unusual case of acute orchitis following aortic surgery. PMID- 12904857 TI - Is prostacyclin an inodilator? PMID- 12904859 TI - Low tidal volume ventilation induces proinflammatory and profibrogenic response in lungs of rats. AB - OBJECTIVE: We examined whether mechanical ventilation with low tidal volume induces polymorphonuclear infiltration and proinflammatory and profibrogenic responses in rat lungs compared dependent and nondependent lung region to expression of interleukin-1beta (IL-1beta) and alpha-1 procollagen III (PC III) mRNA. DESIGN: An experimental, randomized and controlled protocol with previously normal rats. INTERVENTIONS: Three groups of ten animals were studied. Two groups were ventilated (FIO2=0.3) in supine position for 1 h without positive end expiratory pressure, one group with a low tidal volume (6 ml/kg), and the other with a high tidal volume (24 ml/kg). In the third group animals were kept in spontaneous ventilation for 1 h. MEASUREMENTS AND RESULTS: After ventilation the right lung was used to quantify polymorphonuclear infiltration. The left lung was divided into dependent and nondependent regions, and expression of IL-1beta and PC III mRNA was quantified by northern blot analysis. The group ventilated with low tidal volume had greater polymorphonuclear infiltration IL-1beta and PC III mRNA expression than the nonventilated group. Similar results were observed with high tidal volumes. There was no difference between low and high tidal volume ventilation. Expression levels of IL-1beta and PC III mRNA were higher in the nondependent region of ventilated groups and equal in the nonventilated group. CONCLUSIONS: Even a low tidal volume mode of mechanical ventilation induces proinflammatory and profibrogenic response, with a nondependent predominance for IL-1beta and PC III mRNA expression in supine, ventilated, previously normal rats. PMID- 12904858 TI - Influence of the humidification device during acute respiratory distress syndrome. AB - OBJECTIVE: To evaluate the effects of the humidification device on respiratory, hemodynamic and gas exchange parameters in acute respiratory distress syndrome (ARDS) patients. DESIGN: A prospective open study. SETTING: A medical intensive care unit of a university hospital. PATIENTS: Acute respiratory distress syndrome patients, with hypercapnia (PaCO(2)>60 mmHg). INTERVENTION: A progressive reduction of the humidification system dead space (DSh). The following five conditions were tested sequentially: (1). heat and moisture exchanger (internal volume=95 ml) with a tracheal closed-suction system (internal volume=25 ml; total DSh=120 ml), (2). heat and moisture exchanger (internal volume=45 ml) with the closed-suction system (DSh=70 ml), (3). heat and moisture exchanger (internal volume=25 ml) with the closed-suction system (DSh=50 ml), (4). heated humidifier with the closed-suction system (DSh=25 ml) and (5). heated humidifier alone (DSh=0 ml). Recordings were performed at baseline and every 30 min after each artificial dead-space reduction. All ventilatory settings remained constant during the measurement periods. RESULTS: Ten ARDS patients were included. A significant PaCO(2) decrease was observed at each humidification system dead space reduction, compared to baseline: PaCO(2 )=80.3+/-20 mmHg at DSh(120) compared to PaCO(2 )=63.6+/-13 mmHg at DSh(0) ( p<0.05). No changes were observed for hemodynamic and ventilatory parameters between the different humidification devices. CONCLUSION: Artificial airway dead-space reduction allows a significant PaCO(2) reduction. Independently of any respiratory mechanical changes, this very simple maneuver may be of importance when low tidal volume ventilation is used in ARDS patients, and when PaCO(2) lowering is warranted. PMID- 12904860 TI - Endothelial nitric oxide synthase (NOS) is upregulated in rapid progressive pulmonary hypertension of the newborn. AB - OBJECTIVE: To provide evidence for the upregulation of endothelial nitric oxide synthase (eNOS) or inducible nitric oxide synthase (iNOS) in the assumed imbalance in the pathophysiology of rapid progressive pulmonary hypertension of the newborn (RPPHN), which is characterized by abnormal hypertrophy of the pulmonary arterioles and arteries leading to increased pulmonary vascular resistance. Furthermore, to determine the cellular source and topographic distribution of eNOS and iNOS. MATERIAL AND METHODS: Lung biopsies were taken from two term neonates with clinical and echocardiographic evidence of RPPH and of three controls. Biopsies were obtained at an early stage of the disease as well as at post mortem and examined immunohistochemically for the presence of eNOS, iNOS and nitrotyrosine. RESULTS: The endothelial cells of pulmonary arterioles stained significantly for eNOS protein in RPPHN patients. This was not the case in the control infants. There were no differences for nitrotyrosine or iNOS between RPPHN patients and controls. CONCLUSION: Rapid progressive pulmonary hypertension of the newborn leads to compensatory induction of eNOS synthesis specifically in endothelial cells of the pulmonary arterioles. This mechanism of compensation can lead to delayed presentation of RPPHN during the late neonatal period. Exogenous inhaled nitric oxide therapy does not lead to suppression of the endogenous synthesis of nitric oxide. PMID- 12904861 TI - Relationship between injury severity and lactate levels in severely injured patients. AB - OBJECTIVE: To determine the correlation of blood concentration of lactate and severity of injury and survival in severely injured patients. DESIGN AND SETTING: A prospective study of severely injured patients admitted directly from an emergency surgical unit to a surgical intensive care unit with an Injury Severity Score (ISS) of 16 points or more. The study was conducted over 30 months. PATIENTS: 98 severely injured subjects aged between 16 and 82 years with ISS range from 16 to 75 points, overall 25.5% mortality. INTERVENTIONS: Blood lactate concentrations were measured once on admission, twice daily during the first 2 days and once daily during the next 3 days. ISS, Revised Trauma Score, Shock Index, and Trauma and Injury Severity Score were calculated for each subject. MEASUREMENTS AND RESULTS: Of 98 severely injured patients 91 had elevated blood lactate concentration (over 2.0 mmol/l). Regression analyses demonstrated that injury severity, as measured by ISS, can be predicted from lactate concentration on admission, while survival, either actual or predicted by Trauma and Injury Severity Score higher than 0.5, can be predicted from lactate concentration after 12 h. We also found that patients with Shock Index higher than 0.9 had significantly higher lactate levels during the first 36 h than those with values less than 0.9. CONCLUSIONS: This study confirmed the relationship between blood lactate levels and injury severity as well as the prognostic value of blood lactate level for survival of severely injured patients. PMID- 12904863 TI - Quantitative trait loci and molecular markers associated with wheat allelopathy. AB - Wheat ( Triticum aestivum L.) has been examined for allelopathic potential against annual ryegrass ( Lolium rigidum). The bioassay technique, 'equal compartment-agar-method', was employed to evaluate seedling allelopathy in a doubled-haploid (DH) population derived from cv Sunco (weakly allelopathic) and cv Tasman (strongly allelopathic). A significant difference in allelopathic activity was found among the DH lines, which inhibited the root length of ryegrass across a range from 23.7 to 88.3%. The phenotypic data showed that wheat allelopathic activity was distributed normally within this DH population and a substantial transgressive segregation for seedling allelopathic activity was also found. Analysis of restriction fragment length polymorphism (RFLP), amplified fragment length polymorphism (AFLP) and microsatellite (SSRs) markers identified two major QTLs on chromosome 2B associated with wheat allelopathy. The linkage analysis of genetic markers and the QTLs may improve genetic gains for the allelopathic activity through marker-assisted selection in wheat breeding. The development of wheat allelopathic cultivars could reduce the over-reliance of weed control on synthetic herbicides. PMID- 12904862 TI - Cyclic nucleotide phosphodiesterases in pancreatic islets. AB - Cyclic nucleotide phosphodiesterases (PDEs) comprise a family of enzymes (PDE1 PDE11) which hydrolyse cyclic AMP and cyclic GMP to their biologically inactive 5' derivatives. Cyclic AMP is an important physiological amplifier of glucose induced insulin secretion. As PDEs are the only known mechanism for inactivating cyclic nucleotides, it is important to characterise the PDEs present in the pancreatic islet beta cells. Several studies have shown pancreatic islets or beta cells to contain PDE1C, PDE3B and PDE4, with some evidence for PDE10A. Most evidence suggests that PDE3B is the most important in relation to the regulation of insulin release, although PDE1C could have a role. PDE3-selective inhibitors augment glucose-induced insulin secretion. In contrast, activation of beta-cell PDE3B could mediate the inhibitory effect of IGF-1 and leptin on insulin secretion. In vivo, although PDE3 inhibitors augment glucose-induced insulin secretion, concomitant inhibition of PDE3B in liver and adipose tissue induce insulin resistance and PDE3 inhibitors do not induce hypoglycaemia. The development of PDE3 inhibitors as anti-diabetic agents would require differentiation between PDE3B in the beta cell and that in hepatocytes and adipocytes. Through their effects in regulating beta-cell cyclic nucleotide concentrations, PDEs could modulate beta-cell growth, differentiation and survival; some work has shown that selective inhibition of PDE4 prevents diabetes in NOD mice and that selective PDE3 inhibition blocks cytokine-induced nitric oxide production in islet cells. Further work is required to understand the mechanism of regulation and role of the various PDEs in islet-cell function and to validate them as targets for drugs to treat and prevent diabetes. PMID- 12904865 TI - Genetic mapping of the novel Turnip mosaic virus resistance gene TuRB03 in Brassica napus. AB - A new source of resistance to the pathotype 4 isolate of Turnip mosaic virus (TuMV) CDN 1 has been identified in Brassica napus (oilseed rape). Analysis of segregation of resistance to TuMV isolate CDN 1 in a backcross generation following a cross between a resistant and a susceptible B. napus line showed that the resistance was dominant and monogenic. Molecular markers linked to this dominant resistance were identified using amplified fragment length polymorphism (AFLP) and microsatellite bulk segregant analysis. Bulks consisted of individuals from a BC(1) population with the resistant or the susceptible phenotype following challenge with CDN 1. One AFLP and six microsatellite markers were associated with the resistance locus, named TuRB03, and these mapped to the same region on chromosome N6 as a previously mapped TuMV resistance gene TuRB01. Further testing of TuRB03 with other TuMV isolates showed that it was not effective against all pathotype 4 isolates. It was effective against some, but not all pathotype 3 isolates tested. It provided further resolution of TuMV pathotypes by sub dividing pathotypes 3 and 4. TuRB03 also provides a new source of resistance for combining with other resistances in our attempts to generate durable resistance to this virus. PMID- 12904864 TI - Comparative genetics at the gene and chromosome levels between rice ( Oryza sativa) and wildrice ( Zizania palustris). AB - Using comparative genetics, genes, repetitive DNA sequences and chromosomes were studied in the Oryzeae in order to more fully exploit the rice genome sequence data. Of particular focus was Zizania palustris L., n = 15, commonly known as American wildrice. Previous work has shown that rice chromosomes 1, 4 and 9 are duplicated in wildrice. The Adh1 and Adh2 genes were sequenced and, based on phylogenetic analyses, found to be duplicated in wildrice. The majority of the sequence diversity in the Adh sequences was in intron 3, in which were found several MITE insertions. Cytological and molecular approaches were used to analyze the evolution of rDNA and centromeric repetitive sequences in the Oryzeae. In wildrice, copies of the 5S rDNA monomer were found at two loci on two different chromosomes near the centromeres, as in rice. One nucleolar organizer region (NOR) locus was found adjacent to the telomere, as in rice. RCS1, a middle repetitive sequence in rice, was present in all of the centromeres of wildrice. RCS2/CentO, the highly repetitive component of Oryza sativa L. centromeres, was conserved in eight of the Oryza species examined, but was not found in wildrice. Three other middle repetitive centromeric sequences (RCH1, RCH2/CentO and RCH3) were also examined and found to have variable evolutionary patterns between species of Oryza and Zizania. PMID- 12904867 TI - [New understanding of the immunopathogenesis of multiple sclerosis]. AB - Multiple sclerosis is a chronic inflammatory and demyelinating disease of the central nervous system. Although the immune system seems to play an important role in its pathogenesis, target antigens are still uncertain and pathways leading to tissue destruction have not been fully elucidated. Recent studies have significantly contributed to better understanding of the disease process and broadened our view on possible scenarios of disease initiation and progression. Here, we review the role of the immune system in the manifestation and evolution of MS and discuss different pathogenetic concepts. We conclude with an outlook on future strategies for identifying the cause of MS. PMID- 12904866 TI - [Current controversies in the diagnosis and management of cerebral venous and dural sinus thrombosis]. AB - Over the last few years, the results from clinical studies and innovative radiological approaches have significantly altered the management of patients with cerebral venous thrombosis. In contrast to previously held beliefs, cerebral venous thrombosis is considered to be a relatively benign disease with an overall favourable prognosis. Mortality is <10%, and the vast majority of patients recover completely. Although the efficacy of heparin has not been unequivocally proven, it is employed in most centres as the standard therapy. There are not sufficient data supporting long-term anticoagulation. In patients with a proven hypercoagulability syndrome, anticoagulation therapy is generally recommended for a period of 6 months. Whether patients with idiopathic thrombosis should also be treated remains controversial. Non-invasive MR and CT angiography techniques have largely replaced conventional angiography for initial evaluation and follow-up examinations. Both methods have a high sensitivity for cerebral venous thrombosis. Local fibrinolytic therapy or other aggressive recanalizing methods can not be generally recommended. PMID- 12904868 TI - [Electric brain stimulation for epilepsy therapy]. AB - Attempts to control epileptic seizures by electrical brain stimulation have been performed for 50 years. Many different stimulation targets and methods have been investigated. Vagal nerve stimulation (VNS) is now approved for the treatment of refractory epilepsies by several governmental authorities in Europe and North America. However, it is mainly used as a palliative method when patients do not respond to medical treatment and epilepsy surgery is not possible. Numerous studies of the effect of deep brain stimulation (DBS) on epileptic seizures have been performed and almost invariably report remarkable success. However, a limited number of controlled studies failed to show a significant effect. Repetitive transcranial magnetic stimulation (rTMS) also was effective in open studies, and controlled studies are now being carried out. In addition, several uncontrolled reports describe successful treatment of refractory status epilepticus with electroconvulsive therapy (ECT). In summary, with the targets and stimulation parameters investigated so far, the effects of electrical brain stimulation on seizure frequency have been moderate at best. In the animal laboratory, we are now testing high-intensity, low-frequency stimulation of white matter tracts directly connected to the epileptogenic zone (e.g., fornix, corpus callosum) as a new methodology to increase the efficacy of DBS ("overdrive method"). PMID- 12904869 TI - [Cerebrospinal fluid parameters in various stages of HIV infection. Results of cross-sectional and longitudinal analysis]. AB - In up to 50% of all human immunodeficiency virus (HIV) patients, the nervous system is clinically involved. Primary or secondary manifestations of the nervous system have been found in even 90% by neuropathological investigations. We present a retrospective analysis of cerebrospinal fluid (CSF) and serum data of 238 HIV patients. Data of cross-sectional analysis in 208 patients and longitudinal analysis in 30 patients are given. In addition, the viral load in CSF and serum was determined in 29 patients. The HIV patients without opportunistic infections showed increased levels of immunoglobulins and more oligoclonal bands. In later stages of the infection, beta-2 microglobulin as a marker of HIV-associated encephalopathy was increased. In the longitudinal study with an observation period of 1 year, an increase could be observed in total CSF proteins of patients who did not receive antiretroviral treatment. In patients with new opportunistic infections of the central nervous system, similar changes in CSF parameters were noted as in comparison to patients not infected by HIV but with the same opportunistic infections. Analysis of CSF is mandatory for the diagnosis and control of opportunistic infections. PMID- 12904870 TI - [Present state of ultrasonographic diagnosis of temporal arteritis. Results of a prospective study]. AB - Temporal arteritis (TA) is a treatable systemic vasculitis with particular affinity for the temporal and orbital arteries. In a prospective study of 115 patients, we evaluated the diagnostic contribution of duplex sonography of the superficial temporal (STA), okzipital (OCCA), and carotid arteries and Doppler sonography of the supratrochlear artery (DSASTR) in patients suspected of having TA. Forty-seven patients had TA: in the STA, periarterial hypoechogenic tissue (halo) was found in 60% (OCCA 48%) and halo and stenosis in 19% (OCCA 15%) of the TA patients and in 4% of patients with other diseases. The DSASTR was abnormal in 13% of the TA patients. Arteriosclerotic carotid disease was found in 72%, >50% stenosis in 13% of the TA patients. Duplex sonography of the STA reached a diagnostic sensitivity/specificity of 78/96% compared to diagnosis and 91/82% compared to histology. In the STA/OCCA, duplex sonography is a helpful new atraumatic method for the diagnosis of TA, and DSASTR is helpful in detecting TA of the orbital arteries. PMID- 12904871 TI - [Is epilepsy in the elderly overlooked? An investigation in a home for the aged]. AB - The symptoms of epileptic seizures in old patients differ from those in younger patients. Therefore, these seizures may be misinterpreted as symptoms of other "typical" diseases in old people. In an old people's home, we assessed whether a standardized questionnaire is able to reveal undiagnosed epilepsy in the elderly. Reported sudden falls, loss of consciousness, and cramps were the criteria for further diagnostic procedures. We found epilepsy in 11 of 389 study participants. In four of them (1% of the total sample), the epilepsy was newly diagnosed; five more cases remained unclear. Therefore, the total number of epileptic patients might have been even higher. Most of the reported sudden falls and unconsciousness (89%) were due to internal medical or other neurological or orthopaedic causes. Once these have been excluded, the diagnosis of epilepsy should be considered. The results of our pilot study suggest that epilepsy in old people's homes is often unrecognised. Early diagnosis and treatment of epilepsy in the elderly is important to improve prognosis and social consequences for affected persons. PMID- 12904872 TI - [Acute headache with meningism and xanthochromic cerebrospinal fluid.An unusual manifestation of cervical epidural abscess]. AB - We report on a 70-year-old female with acute onset of headache, meningism, xanthochromic cerebrospinal fluid, and developing laboratory parameters indicating a systemic infection. Initially, a subarachnoidal hemorrhage was assumed. However, magnetic resonance imaging showed upper cervical osteomyelitis and extending spinal epidural abscess. After application of broad systemic antibiotics, secluded abscess formation was achieved and successful neurosurgical debridement performed. The unusual cranial concentration of the complaints is probably due to the involvement of the craniocervical transition. Cervical epidural spinal abscess represents a potentially dangerous bacterial infection of the upper spine. Inadequate treatment can lead to irreversible neurological deficits. Cervical magnetic resonance imaging is an effective tool for diagnosis, since early diagnosis is important for good prognosis. PMID- 12904874 TI - [Statin-induced Parkinson's-syndrome. Reader's letter on the article by J. Finsterer in "Der Nervenarzt" (2003) 74:115-122]. PMID- 12904873 TI - [Cholesterol-reducing medications-a new therapeutic option for multiple sclerosis? Statins as immunomodulators]. AB - Statins are inhibitors of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase, which is crucial for cholesterol biosynthesis, and are widely used as lipid-lowering agents. These drugs greatly reduce atherosclerosis and cardiovascular morbidity, which in the past was mainly attributed to their cholesterol-lowering properties. However, recent evidence suggests that statins are also potent immunomodulators. They exerted beneficial effects on animal models of experimental autoimmune encephalomyelitis and thus have therapeutic potential for multiple sclerosis. Their exact mechanism of action is still unclear. HMG-CoA-dependent effects and a direct effect on immune receptors are conceivable and are reviewed here. PMID- 12904875 TI - [Dysfunctions of the stomatognathic system in tinnitus patients compared to controls]. AB - AIMS OF THE STUDY: The high prevalence of ear, nose and throat symptoms in patients with temporomandibular disorders seems to depend on anatomical, functional, neuromuscular, or psychosomatic connections between both phenomena. The present study evaluates the prevalence of temporomandibular dysfunctions in tinnitus patients compared to controls. PATIENTS AND METHODS: A total of 40 patients and 35 controls were clinically examined. The case history as well as the quality of tinnitus and its influence on the patient's life were evaluated, and psychosocial data collected by a questionnaire dealing with stress factors. RESULTS: Tinnitus patients showed muscle tenderness in masticatory muscles as well as in other muscle groups significantly more frequently than controls. The prevalence of arthrogenous dysfunctions was not significantly different between the groups. Occlusal parameters such as instability of intercuspidation, presence of dysgnathia, and signs of parafunctions were significantly more frequent in patients than in controls. The psychosocial assessment showed some trend towards the tinnitus patients feeling more stress in society, daily life, their partnerships and at work. CONCLUSIONS: Tinnitus correlates significantly with myogenous disorders but not with arthrogenous disorders of the temporomandibular joint (TMJ). This result supports the concept of neuromuscular and functional relationships between tinnitus and the stomatognathic system. PMID- 12904877 TI - [Experimental investigations of CO2 laser application in middle ear ossicles]. AB - BACKGROUND: During the last few years, several laser systems have been applied for procedures in middle ear surgery. In this study, we determined the technical parameters for the dissection of the middle ear ossicles with the CO(2) laser and analyzed the histological findings. METHODS: The malleus necks of 16 human temporal bones were dissected under standardized conditions using a CO(2) laser with a power output between 35 and 55 kW/cm(2). The specimens were fixed and histological probes of 50- micro m thickness were prepared. RESULTS: The laser outputs led to crater diameters from 0.14 to 0.55 mm. As an analogy between laser energy and thermal tissue destruction, three zones of thermal damage were differentiated: a cinder zone, a carbonization zone, and a zone of dehydration. The metrical dimensions of these zones did not show any correlation to the applied laser energy. CONCLUSIONS: The data of this study show that commercially available CO(2) lasers are sufficient for a safe and effective partial resection of middle ear ossicles using a power output of 35 kW/cm(2). PMID- 12904876 TI - [Auditory brainstem responses evoked by direct mechanical stimulation of the ossicular chain. Objective preoperative testing of candidates for implantable hearing aids]. AB - BACKGROUND AND OBJECTIVE: The function of several implantable hearing systems is based on the vibratory excitation of the ossicular chain. For preoperative testing, malleus vibration audiometry has been described [7]. In special cases, it could be desirable to perform objective testing before implantation. We therefore studied the auditory brainstem responses (ABR) evoked by vibratory stimulation of the ear drum. METHOD: The ABR evoked by vibratory and acoustical click stimulation were recorded in 20 subjects with normal hearing at four different stimulus intensities. For the vibratory stimulation (v-ABR), the transducer rod was brought into mechanical contact with the eardrum under microscopic control. This setup was further used for the determination of the individual subjective threshold. The v-ABR were compared to conventional ABR recordings. RESULTS: In all subjects, the v-ABR could clearly be identified. At high stimulus levels, the complete wave pattern was observed in 15 of 20 cases. There is no essential difference between v-ABR and conventional ABR with respect to the incidence and parameters of the potentials. At high stimulus levels, a pronounced stimulus artefact impedes the identification of wave I in some cases. Moreover, the quality of v-ABR is limited by the uncomfortable situation and incomplete relaxation of the subject. CONCLUSIONS: The measurement of vibratory evoked ABR is practicable without hazards for the patient and it yields reliable results. The hearing threshold for click stimuli can be determined from the incidence and amplitude of the responses. The loudness associated with the click stimuli can be estimated from the response latency. Therefore, the investigation of v-ABR not only gives evidence of the individual suitability for implantable hearing aids but its outcome may also be useful as an orientation for the postoperative fitting. PMID- 12904878 TI - [Transvestibular closure with an autologous bone graft as surgical repair of a nasoseptal defect due to Le Fort-I osteotomy]. AB - Defects of the nasal septum are a common complication after nasal surgery. Affected patients frequently suffer from bleeding, crusting and impaired nasal air flow. The surgical closure of septal defects remains a distinctive challenge. Though many different techniques have been described, the failure rate of this procedure remains high. In the case presented here, a large basal septum defect occurred after a prosthetic Le Fort-I osteotomy. The attempt to cover the distance between the bony nasal floor and the nasal septum with pedicled mucosal flaps failed due to extensive scar formation of the nasal mucosa. Therefore a nasal floor elevation by insertion of an autologous bone graft from the iliac crest was conducted. The bone graft was connected with the hard palate via two titanium screws. Other than with an autologous cartilage graft, no major resorption of the bone graft is to be expected. This indirect method for the closure of a basal nasal septum defect is new. PMID- 12904879 TI - CT and MR imaging findings of xanthogranulomatous cholecystitis: correlation with pathologic findings. AB - The aim of this study was to evaluate CT and MRI findings in xanthogranulomatous cholecystitis (XGC) and to correlate the imaging findings with various pathologic parameters. The study included 13 patients with histopathologically confirmed XGC. The CT ( n=13) and MRI ( n=5) obtained in these patients were evaluated retrospectively. On CT, low-attenuation areas in the wall of XGC correlated with foam and inflammatory cells or necrosis and/or abscess in XGC. Areas of iso- to slightly high signal intensity on T2-weighted images, showing slight enhancement at early phase and strong enhancement at last phase on dynamic study, corresponded with areas of abundant xanthogranulomas. Areas with very high signal intensity on T2-weighted images without enhancement corresponded with necrosis and/or abscesses. Luminal surface enhancement (LSE) of gallbladder wall represented preservation of the epithelial layer. The early-enhanced areas of the liver bed on dynamic CT and MR images corresponded with accumulation of inflammatory cells and abundant fibrosis. Our results indicate that CT and MRI findings correlate well with the histopathologic findings of XGC. PMID- 12904880 TI - Developmental nasal midline masses in children: neuroradiological evaluation. AB - Developmental nasal midline masses in children are rare lesions. Neuroimaging is essential to characterise these lesions, to determine the exact location of the lesion and most importantly to exclude a possible intracranial extension or connection. Our objective was to evaluate CT and MRI in the diagnosis of developmental nasal midline masses. Eleven patients (mean age 4.5 years) with nasal midline masses were examined by CT and MRI. Neuroimaging was evaluated for (a) lesion location/size, (b) indirect (bifid or deformed crista galli, widened foramen caecum, defect of the cribriform plate) and direct (identification of intracranially located lesion components or signal alterations) imaging signs of intracranial extension, (c) secondary complications and (d) associated malformations. Surgical and histological findings served as gold standard. Nasal dermoid sinus cysts were diagnosed in 9 patients. One patient was diagnosed with an meningocele and another patient with a nasal glioma. Indirect CT and MRI signs correlated with the surgical results in 10 of 11 patients. Direct CT findings correlated with surgery in all patients, whereas the direct MRI signs correlated in 9 of 11 patients. In 2 patients MRI showed an intracranial signal alteration not seen on CT. Neuroimaging corrected the clinical diagnosis in 1 patient. One child presented with a meningitis. In none of the patients was an associated malformation diagnosed. Intracranial extension is equally well detected by CT and MRI using indirect imaging signs. Evaluating the direct imaging signs, MRI suspected intracranial components in 2 patients without a correlate on CT. This could represent an isolated intracranial component that got undetected on CT and surgery. In 9 patients CT and MRI matched the surgical findings. The MRI did not show any false-negative results. These results in combination with the multiplanar MRI capabilities, the different image contrasts that can be generated by MRI and the lack of radiation favour the use of MRI as primary imaging tool in these young patients in which the region of imaging is usually centred on the radiosensitive eye lenses. PMID- 12904883 TI - Diffusion-weighted MR imaging of pleural fluid: differentiation of transudative vs exudative pleural effusions. AB - The aim of this study was to evaluate the ability of diffusion-weighted MRI in differentiating transudative from exudative pleural effusions. Fifty-seven patients with pleural effusion were studied. Diffusion-weighted imaging (DWI) was performed with an echo-planar imaging (EPI) sequence (b values 0, 1000 s/mm(2)) in 52 patients. The apparent diffusion coefficient (ADC) values were reconstructed from three different regions. Subsequently, thoracentesis was performed and the pleural fluid was analyzed. Laboratory results revealed 20 transudative and 32 exudative effusions. Transudates had a mean ADC value of 3.42+/-0.76 x 10(-3) mm(2)/s. Exudates had a mean ADC value of 3.18+/-1.82 x 10( 3) mm(2)/s. The optimum cutoff point for ADC values was 3.38 x 10(-3) mm(2)/s with a sensitivity of 90.6% and specificity of 85%. A significant negative correlation was seen between ADC values and pleural fluid protein, albumin concentrations and lactate dehydrogenase (LDH) measurements ( r=-0.69, -0.66, and -0.46, respectively; p<0.01). The positive predictive value, negative predictive value, and diagnostic accuracy of ADC values were determined to be 90.6, 85, and 88.5%, respectively. The application of diffusion gradients to analyze pleural fluid may be an alternative to the thoracentesis. Non-invasive characterization of a pleural effusion by means of DWI with single-shot EPI technique may obviate the need for thoracentesis with its associated patient morbidity. PMID- 12904884 TI - Toward comparability of coronary magnetic resonance angiography: proposal for a standardized quantitative assessment. AB - A method is proposed for the quantitative assessment of coronary magnetic resonance angiography (MRA) acquisitions. The method is based on four parameters: signal-to-noise ratio (SNR); contrast-to-noise ratio (CNR); vessel length; and vessel-edge definition. A pig model (n=7) was used to illustrate the proposed quantitative analysis method. Three-dimensional gradient-echo coronary MRA was performed with and without exogenous contrast enhancement using a gadolinium based blood-pool contrast agent (Vistarem, Guerbet, Aulnay-Sous-Bois, France). The acquired images could be well differentiated based on the four parameters. The SNR was calculated as 9.0+/-1.4 vs 10.4+/-2.1, the CNR as 6.2+/-0.8 vs 8.2+/ 0.9, the vessel length as 48.2+/-11.6 vs 86.5+/-13.8 mm, and the vessel-edge definition as 4.9+/-1.5 vs 7.7+/-3.4. Different coronary MRA techniques can be evaluated objectively with the combined use of SNR, CNR, vessel length, and vessel-edge parameters. PMID- 12904885 TI - Measurement of cardiac output from a test-bolus injection in multislice computed tomography. AB - The aim of this study was to assess the feasibility of non-invasive determination of cardiac function from test-bolus data in multislice spiral computed tomography (MSCT). In 25 patients enhancement data gathered from a standardized test-bolus injection were analyzed. The test-bolus examination was performed prior to a retrospectively ECG-gated MSCT of the heart. A time-attenuation curve was obtained in the ascending aorta at the level of the pulmonary arteries. A gamma variate fit was applied to the curve in order to exclude recirculation and get pure first-pass data. Using the known amount of iodine injected, cardiac output (CO), and stroke volume (SV) were determined from integration of the fitted contrast enhancement curve using a reformation of the Stewart-Hamilton equation. Results were compared with CO and SV calculated from the geometric analysis of the retrospectively gated MSCT data using the ARGUS Software (Siemens, Forchheim, Germany). The CO and SV determined from test-bolus analysis and from geometric analysis correlated well with Pearson's correlation coefficients of 0.87 and 0.88, respectively. The standard deviation of the difference between both methods was 0.51 l/min for CO (8.6%) and 11.0 ml for SV (12.3%). Non-invasive quantification of CO seems to be feasible from a standard test-bolus injection. It provides valuable information on cardiac function without additional radiation or application of contrast material. PMID- 12904886 TI - An effective method for axillary bud culture and RAPD analysis of cloned plants in tetraploid black locust. AB - An efficient micropropagation technique by axillary bud multiplication was established for cloning tetraploid black locust tree (Robinia pseudoacacia L.). The result showed that the optimal medium for shoot multiplication and elongation was Murashige and Skoog (MS) medium supplemented with 0.5 mg/l 6 benzylaminopurine in combination with 0.5 mg/l kinetin and 0.1 mg/l 1 naphthaleneacetic acid. The best medium for rooting was half-strength MS medium with 0.25 mg/l indole-3-butyric acid. In the present report, we examined the genetic fidelity of the micropropagated plants by the randomly amplified polymorphic DNA (RAPD) method with 25 primers. The cloned plants of tetraploid black locust showed complete stability. PMID- 12904887 TI - Rheumatoid arthritis. Introduction. PMID- 12904888 TI - Immunopathogenesis of collagen arthritis. AB - Collagen-induced arthritis (CIA) is an animal model of autoimmunity that has been studied extensively because of its similarities to rheumatoid arthritis (RA). CIA is induced in genetically susceptible strains of mice by immunization with type II collagen (CII), and both T cell and B cell immunity to CII are required for disease manifestation. Like RA, CIA is primarily an autoimmune disease of articular joints and susceptibility to CIA is linked to specific class II molecules of the major histocompatibility complex (H-2(r) and H-2(q)). Recently, it was demonstrated that transgenic expression of HLA-DR1 (*0101) or DR4 (*0401) molecules associated with susceptibility to RA also conferred susceptibility to CIA in the mouse model. The T cell response to CII has been extensively characterized in both the DR transgenic and naturally susceptible mouse strains, including the antigenic determinants recognized, the role of post transcriptional modifications of these determinants in the pathogenic T cell response, and the cytokines produced. Like most class II-mediated autoimmune diseases, the cytokine production of CII-specific T cells reflects a Th1 phenotype of the autoimmune response. While the direct role of T cells in the pathogenesis of CIA is unclear, the B cell response in terms of anti-CII immunoglobulin is critical to the development of the disease. This response, predominated by the IgG2 isotype, requires the activation of the complement cascade for the development of CIA. In recent years, the pathogenesis of this model has been studied extensively and the CIA model is proving to be a valuable asset for the design of new immunotherapeutics for the potential treatment of RA and other autoimmune diseases. PMID- 12904889 TI - The TNF-alpha transgenic mouse model of inflammatory arthritis. AB - Rheumatoid arthritis is a chronic inflammatory disorder that affects multiple peripheral joints. It is the most common form of inflammatory arthritis and is characterized by synovial hyperplasia, immune cell infiltration, cartilage destruction, and bone erosion. To gain insight into the etiology of the disease, a variety of animal models have been established. Twelve years ago George Kollias' laboratory generated a transgenic (Tg) mouse that over-expresses human TNF-alpha, and develops an erosive polyarthritis with many characteristics observed in rheumatoid arthritis patients. The phenotype of this mouse model validated the theory that TNF-alpha is at the apex of the pro-inflammatory cascade in rheumatoid arthritis, and foreshadowed the remarkable success of anti TNF-alpha therapy that has transformed the effective management of this disease. As such, the TNF-Tg mice are very useful tools for dissecting the molecular mechanisms of the pathogenic process and evaluating the efficacy of novel therapeutic strategies for rheumatoid arthritis. In this review we (1) provide a brief summary of TNF-alpha biology and the role of this dominant cytokine in rheumatoid arthritis, (2) describe the various TNF-Tg models and their phenotypes, and (3) give examples of how this model has been used experimentally. PMID- 12904890 TI - Development of humanized mice as a model of inflammatory arthritis. AB - Clinically, rheumatoid arthritis (RA) is a chronic deforming disease characterized mainly by joint swelling and destruction. Although synovial inflammation and bone erosion are the hallmarks of this disease, the presentation of various features between patients is clearly heterogenous, suggesting that there are different variants of RA. Hence, the development of an animal model that has all of the elements of human RA has remained elusive. This review explores several different views on the etiology of RA and the recent data from various murine arthritis models which provide support for these theories. In addition to discussing the potential roles of CD4(+) T cell activation, autoantibodies, and lymphocyte-independent cytokine production, the role of CD4(+) T regulatory cells will be presented in the context of a newly developed humanized transgenic mouse model. This novel T cell receptor transgenic model is being characterized to enhance our understanding of the mechanisms involved in the breach of self-tolerance that occurs in autoimmune disorders such as RA. PMID- 12904891 TI - Heat shock protein 60 and adjuvant arthritis: a model for T cell regulation in human arthritis. AB - Heat shock proteins (hsp) are highly conserved, immune-dominant microbial proteins, whose expression is increased at sites of inflammation. In the experimental model of adjuvant arthritis (AA) immune responses to hsp determine the outcome of disease. AA can be transferred with a single T cell clone specific for a sequence of mycobacterial hsp65 (Mhsp65). Immunization with whole Mhsp65 on the other hand, protects in virtually all forms of experimental arthritis, including AA. This protective effect seems the consequence of the induction of a T cell response directed against self-hsp60. A similar protective effect of self hsp60-specific T cells seems present in patients with a spontaneous remitting form of juvenile idiopathic arthritis. Next to hsp60, other hsp have similar protective effects in arthritis, while other conserved microbial proteins lack such capacity. Nasal administration of hsp60 peptides induces IL-10-driven regulatory T cells that are highly effective in suppressing arthritis. Thus hsp60, or peptides derived from hsp60, are suitable candidates for immune therapy in chronic arthritis. PMID- 12904892 TI - The SCID mouse model: novel therapeutic targets - lessons from gene transfer. AB - The hallmark of rheumatoid arthritis (RA) is progressive destruction of the joints, preceded and accompanied by synovial hyperplasia and chronic inflammation. Spontaneous and induced animal models of RA reflect predominantly the inflammatory aspects of the disease. To reproduce the destruction of cartilage and bone mediated by an activated synovium, it was desirable to develop models that allow the dissection of cellular and molecular components derived from human tissue. The SCID mouse co-implantation model of human RA focuses on RA synovial fibroblasts (RA-SF) and their role in cartilage destruction. The model has provided the best evidence that RA-SF contribute significantly to matrix degradation, even in the absence of human lymphocytes and macrophages, since highly purified RA-SF invade the co-implanted normal human cartilage. Moreover, it became clear that they maintained their aggressive phenotype over long periods of time, particularly at sites of invasion into the co-implanted human cartilage. Targeting different signaling molecules, cytokines and matrix-degrading enzymes by soluble receptors, antagonists or negative mutants in the SCID mouse model of RA has implicated many of them in the mechanisms leading to cartilage destruction. However, since inhibition of a single molecule or pathway is not sufficient to inhibit the aggressive behavior of RA-SF it appears necessary to co express in the synoviocytes genes for two or even more antagonists of e.g. cytokines, matrix-degrading enzymes or molecules interfering specifically with signaling pathways involved in the apoptosis of RA-SF. Based on the recent observation that the L1 (line-1) endogenous retroviral element appears responsible for the cytokine- independent activation via the MAPK p38delta, the current understanding of disease pathogenesis suggests that both the cytokine dependent as well as the cytokine-independent pathways of joint destruction must be inhibited. Modulation of both pathways by gene transfer approaches in the SCID mouse model is a feasible method aimed at identifying novel targets for the prevention of cartilage destruction in RA. PMID- 12904893 TI - The KRN mouse model of inflammatory arthritis. AB - In 1996 a new murine model of spontaneous arthritis was described by the group of Benoist and Mathis. Mice transgenic for a T cell receptor recognizing an epitope of bovine RNase and bred onto a NOD background developed severe destructive arthritis, which resembles human rheumatoid arthritis in many respects. The development of disease requires the presence of T and B lymphocytes and is dependent on the MHC class II molecule I-A(g7). B cell activation by antigen and an additional CD40-CD40 ligand interaction was found to give rise to the production of autoantibodies. Glucose-6-phosphate isomerase was identified as the target of the autoantibodies; moreover, the transgenic T cells were demonstrated to exhibit a dual specificity for both bovine RNase and glucose-6-phosphate isomerase. Importantly, the arthritis is serum transferable to normal recipients, enabling the examination of the pathogenic mechanisms of joint inflammation and destruction. Recent studies suggest the crucial involvement of the innate immune system in the development of antibody-induced arthritis. Complement components, Fc receptors and neutrophils are indispensable for disease induction. An overview of the existing data is given and the emerging concepts of the pathogenesis of the K/BxN arthritis are discussed with respect to their relevance for human rheumatoid arthritis. Because of the reliable and robust induction of joint inflammation by serum transfer this new disease model has been and will be a valuable means to address the as-yet-unanswered key questions related to the development of arthritis. PMID- 12904894 TI - Dihydropyrimidine dehydrogenase (DPD) rapidly regenerates after inactivation by eniluracil (GW776C85) in primary and metastatic colorectal cancer. AB - PURPOSE: Catabolism of 5-fluorouracil (5-FU) is primarily regulated by DPD. Inactivation of DPD using eniluracil is advantageous in that it renders 5-FU orally bioavailable with more predictable pharmacokinetics and blocks one of the major potential mechanisms of 5-FU chemoresistance. The purpose of this study was to initially document inactivation of DPD by eniluracil in primary and metastatic colorectal cancer (CRC) and then to assess the time-course of the regeneration of DPD activity in peripheral blood (and where possible, additional tissues). METHODS: Of 28 patients entered, 23 were randomized to preoperative oral eniluracil (20 mg orally twice daily) or placebo prior to definitive resection of primary or metastatic CRC. Three patients were replaced, two because they had no residual tumor on pathologic evaluation and one for not taking the study drug. Patients received eniluracil 48, 36, 24 and approximately 12 h prior to surgical resection. In a second part of the study to document tissue regeneration of DPD, the additional five patients received eniluracil 144, 132, 120 and 108 h prior to surgical resection. DPD activity was measured in normal tissues, tumors and peripheral blood mononuclear cells (PBMC). Serum eniluracil and plasma uracil concentrations were determined before and through 28 days after eniluracil dosing. Data are presented as means+/-SEM, and significance defined as P<0.05. RESULTS: Eniluracil inactivated DPD below the level of detection in primary and metastatic CRC as well as in normal tissues (0.0 pmol/min per mg protein) compared to primary tumor, metastatic tumor, PBMC, normal mucosa, and normal liver of patients receiving placebo (57+/-12, 119+/-19, 157+/-22, 77+/-12, 243+/ 24 pmol/min/mg protein, respectively; P<0.05). At the time of surgery, serum eniluracil and uracil concentrations were 207+/-36 ng/ml and 2700+/-170 ng/ml in drug-treated patients. Within 6 days following treatment with eniluracil, serum eniluracil and uracil concentrations were undetectable, while DPD activity in PBMC had returned to baseline. The second group of patients (n=5) were given eniluracil 8 and 7 days prior to surgery to evaluate DPD regeneration in normal tissues and primary CRC tissue. In samples of these tissues, collected 6 days after the last eniluracil dose, DPD activity approached baseline in normal mucosa, normal liver and primary tumor (28+/-12, 94+/-23 and 20+/-8 pmol/min per mg protein, respectively). CONCLUSIONS: These results demonstrate that oral administration of eniluracil inactivated DPD below the level of detection in normal tissues as well as in primary and metastatic CRC. After discontinuation of eniluracil, DPD rapidly returned toward baseline within 6 days in PBMC, normal intestinal mucosa and normal liver. PMID- 12904895 TI - Alleviation of side effects induced by irinotecan hydrochloride (CPT-11) in rats by intravenous infusion. AB - PURPOSE: Irinotecan hydrochloride (CPT-11) is a potent topoisomerase I inhibitor and is established and used widely as an antitumor agent. However, it sometimes causes severe side effects such as myelosuppression and diarrhea. These dose limiting toxicities prevent the adoption of CPT-11 in aggressive chemotherapy. Thus we sought to determine in a rat model whether extending the period of infusion of CPT-11 would ameliorate the adverse reactions. METHODS: CPT-11 was administered intravenously (i.v.) to rats at a dose of 60 mg/kg per day for four consecutive days as a bolus injection or as 3-, 8- or 24-h infusions, and then blood cell counts and the incidence of acute and delayed-onset diarrhea were monitored. RESULTS: Serious acute and delayed-onset diarrhea and marked decreases in the number of neutrophils and lymphocytes were observed in the bolus injection group. These symptoms were alleviated in the infusion groups with the degree of alleviation dependent on infusion time. In the bolus injection group, mucosal impairment of the cecal epithelium including wall thickening, edema, a decrease in the number and size of crypts, and the formation of a pseudomembrane-like substance was observed, whereas these changes were less severe in the infusion groups. Areas under the plasma concentration-time curves (AUCpla) of CPT-11 and its metabolite, 7-ethyl-10-hydroxycamptothecin (SN-38), differed little among the bolus injection group, and the 3-h and 8-h infusion groups. However, the AUCpla values of CPT-11 and SN-38 were significantly decreased and increased, respectively, in the 24-h infusion group. The maximum plasma concentrations (Cmax) of CPT-11 decreased with increasing infusion time, but those of SN-38 did not. CONCLUSIONS: It was confirmed that the side effects of CPT-11 were alleviated by extending the infusion time. The pharmacokinetic parameters suggested that the Cmax of CPT-11 is closely related to the incidence and severity of adverse reactions such as myelosuppression and acute and delayed onset diarrhea. PMID- 12904896 TI - Phase I trial of perillyl alcohol administered four times daily continuously. AB - PURPOSE: Previous experience with perillyl alcohol (POH) was with a formulation of 500-mg capsules each containing 250 mg POH and soybean oil. This formulation resulted in the ingestion of large amounts of soybean oil (>10 g/day). Dose limiting toxicities (DLT) were primarily gastrointestinal. Prior studies also showed no further increase in POH metabolite concentrations with doses of >1600 mg/m2. Therefore, a new formulation of POH was developed (700 mg containing 675 mg POH) in an effort to improve dose and metabolite concentrations delivered and toxicity encountered with chronic dosing. EXPERIMENTAL DESIGN: Eligible patients had refractory solid malignancies. Dose escalation occurred in cohorts of three at the dose levels/dose of 1350 mg, 2025 mg, 2700 mg, 3375 mg and 4050 mg, administered orally four times a day in a 28-day cycle. RESULTS: A group of 19 patients were enrolled. One DLT occurred at dose level 5. This cohort was expanded to six patients, and no further DLT occurred. The maximum tolerated dose was not reached. The predominant toxicity was gastrointestinal. Nausea and vomiting occurred in 63% of patients (12/19, grade 1 in 10). The same proportion of patients (12/19) experienced heartburn and indigestion, primarily grade 1. Although the side effects were mild in nature, three patients withdrew from treatment, citing intolerable gastrointestinal toxicity. The AUCs of POH metabolites did not appear to increase from level 1 to level 2 or change significantly from day 1 to day 29. Inter- and intrapatient variability in metabolite levels was observed. CONCLUSIONS: This reformulation of POH appears to be an improvement upon the prior formulation, by reducing the number of capsules ingested and the degree of gastrointestinal toxicity per dose. It does not appear to offer any metabolite pharmacokinetic advantage. A dose of 2050 mg administered four times daily was easily tolerated. Higher doses can be administered but with increasing gastrointestinal toxicity that limits compliance. PMID- 12904897 TI - Assessment of normal and tumor tissue uptake of MAG-CPT, a polymer-bound prodrug of camptothecin, in patients undergoing elective surgery for colorectal carcinoma. AB - PURPOSE: MAG-camptothecin (MAG-CPT) is the lead compound of a novel drug delivery system in which an active cytotoxic moiety, camptothecin (CPT), is covalently linked to a soluble polymeric carrier (MAG) to form an inactive prodrug. The mechanism of action of CPT remains unaltered, but the delivery system is thought to allow the carrier-bound drug to accumulate in tumor tissues and release the active CPT locally. This proof-of-concept clinical study was designed to determine whether MAG-CPT was preferentially delivered to or retained in tumor tissue compared to adjacent normal tissue or plasma, and to estimate the degree of intratissue release of CPT. METHODS: This was an open, non-randomized study in ten adult patients scheduled for elective surgery for colorectal cancer. Patients received a single dose of 60 mg/m2 (CPT equivalent) of MAG-CPT 24 h, 3 days or 7 days prior to surgery. Plasma, tumor, and adjacent normal tissue samples were collected simultaneously at the time of surgery and analyzed for MAG-bound and released CPT concentrations. RESULTS: MAG-bound and free CPT concentrations in plasma, tumor, and normal tissue achieved equilibrium by 24 h after dosing, declining in parallel up to 7 days after dosing. MAG-bound CPT was delivered to similar levels to tumor and normal tissue. At 24 h after dosing, the mean+/-SD MAG-bound CPT concentrations were 861+/-216 ng/g in tumor and 751+/-215 ng/g in adjacent normal tissue, and free CPT concentrations were lower in tumor than in normal tissue (12.2+/-4.7 ng/g and 21.9+/-6.7 ng/g, respectively). At 24 h after dosing, mean+/-SD ratios of MAG-bound and free CPT in tumor and plasma were 0.13+/-0.03 and 0.22+/-0.09, respectively, and the ratios did not change for up to 7 days after dosing, indicating a lack of preferential retention of MAG-bound CPT or release of free CPT in tumor. These results are in marked contrast to previous data from animal tumor xenograft studies, where MAG-CPT levels were higher in tissue than in plasma at 3 and 7 days after a single i.v. dose. CONCLUSIONS: Delivery of CPT to the target tumor tissue is achievable by means of the MAG-CPT polymer-bound delivery system, with the equilibrium between plasma and tumor tissue concentrations of released CPT being established within 24 h after dosing. However, preferential retention of MAG-bound or released CPT in the tumor relative to normal tissue or plasma was not detected during the 7 days after dosing. The methods employed in our study could be of use in making "go/no go" decisions on further development of anticancer drugs. PMID- 12904898 TI - Safety of lumbar puncture for adults with acute leukemia and restrictive prophylactic platelet transfusion. AB - No data exist on the trigger for platelet transfusions in adult thrombocytopenic patients with acute leukemia undergoing lumbar puncture (LP). We reviewed the records of 66 patients with acute leukemia (median age 38 years, range 18-68) who have been treated in our institution for 6 years. A total of 195 LPs were performed. No serious hemorrhagic complications occurred, but there was a significant trend towards a higher percentage of traumatic procedures, defined as the occurrence of >500 erythrocytes per high-power field, in patients with lowest platelet counts ( p<0.005). Although not associated with serious clinical bleeding events in this study, the increased occurrence of traumatic procedures may indicate an increased risk for more serious hemorrhagic complications, implying a trigger not lower than 20x10(9)/L for prophylactic transfusions of platelets in adult patients with acute leukemia undergoing LP. PMID- 12904899 TI - Decreased expression of membrane alpha4beta1, alpha5beta1 integrins and transferrin receptor on erythroblasts in splenectomized patients with beta thalassemia intermedia. Parallel assessment of serum soluble transferrin receptors levels. AB - Dysfunction of cell membrane is a recognized consequence of the pathogenetic process underlying the beta-thalassemia syndromes and it is reasonable to hypothesize that surface structures crucial for the development of erythroid lineage may also be affected. The study included six adult splenectomized patients with beta-thalassemia intermedia. Expression of alpha4beta1 integrin (CD49d/CD29), alpha5beta1 integrin (CD49e/CD29) and transferrin receptor (CD71) on peripheral blood and bone marrow erythroblasts and on erythroid precursors grown in vitro was studied by flow cytometry and immunocytochemistry. Serum soluble transferrin receptor levels (sCD71) were also measured with enzyme-linked immunosorbent assay. In beta-thalassemic patients, significant reduction of CD49d, CD29 and CD71 expression was found in peripheral blood nucleated red cells, compared to patients presenting with erythroblasts in the circulation because of other diseases. Marrow erythroblasts were also deficient for the same molecules against the erythroblasts in iron deficiency anemia. All molecules tested were greatly diminished on erythroid precursors developed in vitro from the patients' cells. Serum sCD71 levels were much higher in thalassemic patients in comparison to both patients with iron deficiency anemia and healthy individuals. The loss of certain integrins and CD71 from erythroid precursors in beta-thalassemia intermedia could be attributed to a generalized membrane dysfunction, perhaps affecting the integrity of their transmembrane domains. The elevation of serum sCD71 levels may be the result of the increased red cell lineage turnover or, alternatively, may indicate increased shedding from the cells to prevent iron overload. In any case, further molecular study of the membrane components is warranted to provide a better understanding of the pathogenetic process in beta-thalassemia syndromes. PMID- 12904900 TI - Increased cell apoptosis in bone marrow trephine biopsies and immunomagnetically isolated myeloid progenitor cells in patients with chronic idiopathic neutropenia. AB - The frequency of apoptotic cells in bone marrow trephine biopsies and cytospins of immunomagnetically isolated myeloid progenitor cells was determined in 39 patients with chronic idiopathic neutropenia (CIN) and 12 hematologically normal individuals using the in situ end-labeling (ISEL) apoptosis detection method. We found that 66.7% of the patients but none of the normal controls displayed apoptotic cells equal to or higher than 5% of the total mononuclear cells in bone marrow biopsies (p<0.01). In the double stain, we also found that the proportion of apoptotic CD15(+) myeloid precursor cells did not differ significantly between patients and control subjects, while the proportion of apoptotic CD34(+) hemopoietic cells could not be estimated with accuracy because of the presence of CD34(+) endothelial cells. Significantly increased apoptosis was noted in cytospins of immunomagnetically isolated patient CD34(+) and CD34(+)/CD33(+) cells but not CD34(-)/CD33(+) cells, compared to the controls ( p<0.001, p<0.02 and p>0.05, respectively). These findings confirm and extend our previous observations in flow-cytometric studies of apoptosis in CIN, indicating that increased apoptosis in CIN bone marrow concerns mainly the CD34(+) and CD34(+)/CD33(+) progenitor cell compartments. We conclude that the accelerated apoptosis in these compartments may account for the impaired neutrophil production in CIN patients. PMID- 12904901 TI - Subtotal splenectomy for the treatment of chronic lymphocytic leukemia. AB - Although splenectomy is helpful in the management of selected patients with chronic lymphocytic leukemia (CLL), in most cases this procedure is accompanied by a greater morbidity and mortality, mainly due to sepsis. Thus, it may be proposed that a conservative procedure that reduces the spleen size may have an effect similar to that of total spleen ablation for the treatment of CLL. The present paper describes our experience with an 81-year-old patient submitted to subtotal splenectomy for treatment of CLL. Indications for surgery were uncontrolled leukemic activity and intense abdominal discomfort due to the huge spleen. The good results obtained with subtotal splenectomy in the present case indicate that this procedure may be a new alternative for the treatment of CLL when removal of the spleen is indicated. PMID- 12904902 TI - The sympathetic contributions to the cardiac plexus. AB - Cardiac sympathetic denervation for intractable angina pectoris in patients unsuitable for conventional revascularization is currently gaining popularity since this procedure may be performed via minimally invasive surgery. A thorough understanding of cardiac innervation and its variations is crucial to successfully effect cardiac denervation. This study aimed to demonstrate the cervical and thoracic sympathetic contributions to the cardiac plexus. The cervical and thoracic sympathetic trunks in 21 fetuses and eight adults were micro-dissected bilaterally and documented ( n=58 sides). The superior cervical cardiac ramus originated from the superior cervical ganglion (present in all specimens) in 53% of cases. The middle cervical ganglion (incidence 81%) gave rise to the middle cervical cardiac ramus in 88% of cases. The cervico-thoracic ganglion (incidence 85%) gave the cervico-thoracic cardiac ramus in 84%. In the thoracic region, four cardiac rami arose from the T2-T6 segment of the thoracic sympathetic trunk. All cervical and thoracic cardiac rami were traced consistently to the deep cardiac plexus. Khogali et al.'s (1999) success of limited T2-T4 sympathectomy in relieving pain at rest of patients with intractable angina pectoris appears to indicate that a significant afferent pain pathway from the heart is selectively interrupted. The variability in pattern of the cervical ganglia, cardiac rami and cervical contributions to the cardiac plexus does not appear to affect the outcome of limited sympathectomy. The complexity of cardiac pain pathways is not fully understood. The study is continuing and attempts to contribute to defining these cardiac neuronal pathways. PMID- 12904904 TI - Intracapsular hip fractures in patients with rheumatoid arthritis. AB - We reviewed the treatment of 43 patients with rheumatoid arthritis and femoral neck fracture. Patients' average age was 66.4 (36-80) years and average duration of RA was 20.3 (4-42) years. Thirteen patients were treated with primary total hip arthroplasty (THA), and the clinical results were comparable to patients treated conservatively or by osteosynthesis. Eighteen patients were treated with primary bipolar hip prosthesis and after an average of 6.1 (1-13) years there was no acetabular destruction. However, long-term results were inferior to patients treated with THA. Nine patients were treated with osteosynthesis, of which two later had a hip prosthesis. Three cases with impacted fractures were treated conservatively with successful union in all. PMID- 12904903 TI - A cadaveric study of a combined trans-mandibular and trans-zygomatic approach to the infratemporal fossa. AB - The infratemporal fossa (ITF) is a deep retromaxillary space corresponding to the inferior aspect of the middle cranial fossa. There are numerous surgical approaches, indicating the difficulty of access, accentuated by the communications with the neighboring regions. The aim of this anatomical study was the optimization and photographic demonstration of the pre-auricular infra temporal trans-zygomatic approach combined with a trans-mandibular approach allowing access to the different regions of the ITF. Six human heads preserved in formalin were dissected under the operating microscope (magnification between x6 and x4). Intra-arterial injection of colored silicone had been done in advance. A plastinated hemi-head was conserved as a teaching and training specimen using the S10 silicone technique. After a pre-auricular temporo-cervical incision, skin flaps, the superficial temporalis fascia and the temporal fascia were elevated. The facial nerve and its branches were preserved and the temporal and zygomatic branches were protected by the superficial temporalis fascia. The division of the temporal muscle and osteotomy of the zygomatic arch opened the superior part of the ITF. Osteotomy of the mandibular ramus preserving the division of the mandibular nerve opened the inferior part of the ITF. This was thus perfectly exposed and access was not impeded by the trunk of the facial nerve and its branches. The place of this complex approach is discussed among the principal exocranial surgical approaches to the ITF. PMID- 12904905 TI - Arthroscopic revisions in failed meniscal surgery. AB - The aim of this study was to classify meniscal tear forms as found in 195 revision arthroscopies. Interval between primary arthroscopy and revision was 7.8+/-5.6 month. All patients were available for control after 1 year. In 174 knees the lesion was located in the medial meniscus and in 21 knees in the lateral meniscus. In the medial meniscus an unstable posterior meniscal horn was seen in 93 knees followed by incomplete horizontal tear and meniscal destruction in 37. Flap tear, circumferential tear, and failed meniscal repair were also seen. In the lateral meniscus destruction of a discoid meniscus, instability near the popliteal hiatus, and various tear forms were seen with nearly equal frequency. Postoperatively Lysholm score increased significantly in both groups. Most meniscal tears, found in revision arthroscopy, are caused by an insufficient primary operation. A diligent analysis of the tear form is absolutely necessary. An adequate radical resection technique to establish a smooth meniscal crest is indispensable. PMID- 12904906 TI - The International Documentation and Evaluation System (IDES)--10-years experience. PMID- 12904909 TI - Primary clear cell sarcoma (malignant melanoma) in the right radius. AB - Clear cell sarcoma (malignant melanoma) originating in bone is an extremely rare occurrence, which has been reported twice previously. It is challenging to differentiate this neoplasm from skeletal metastasis of malignant melanoma because it shows no specific imaging, pathological or immunohistochemical features. However, this differentiation is clinically important due to significant differences in patient management. In this article, we present the case of a 55-year-old man with primary clear cell sarcoma arising in the right radius. PMID- 12904910 TI - The conformational states of Mg.ATP in water. AB - The conformational equilibria of Mg.ATP in solution is studied using molecular dynamics (MD) augmented with umbrella sampling methods. Free energy comparisons show that the Mg(2+) ion is equally likely to coordinate the oxygens of the two end phosphates, or of all three phosphates. The MD trajectories reveal two major degrees of freedom of the Mg.ATP molecule in solution, and we compute the free energy as a function of these variables, and determine its elastic properties. Comparing the free energy function with several crystallographic structures of ATP analogs, we find that the crystal structures correspond to states where ATP would be elastically strained. The average water density around Mg.ATP is investigated to show the average number of hydrogen bonds and the hydrophobicity. PMID- 12904911 TI - Characterisation of germinating and non-germinating wheat seeds by nuclear magnetic resonance (NMR) spectroscopy. AB - Experiments were conducted to characterise the changes, especially of water status in germinating and non-germinating wheat seeds by nuclear magnetic resonance (NMR) spectroscopy. NMR relaxation time ( T(2)) measurements showed tri phasic or bi-phasic characteristics during different stages of hydration, depending on the seed's ability to germinate. Component analysis of T(2) data revealed the existence of only two components, bound and bulk water, in dry seeds. In contrast, both the germinating and non-germinating wheat seeds had a three-component water proton system (bound, bulk and free water) in phase I of hydration. During the lag phase (phase II) of hydration, bulk water component of non-germinating seeds disappeared completely, resulting in a two component water proton system. Nevertheless, the three component water proton system was observed in the germinating seeds in phase II. Following phase II, rapid hydration (phase III) was observed in germinating seeds only. Water protons were re-organised and there were increases in bulk and free water but decreases in bound water concomitantly. Comparison of the physical state of water in these seeds by NMR spectroscopy with that of tissue leachate conductivity measurement suggests that the seed membrane system was affected more evidently in non-germinating seeds, leading to the disorganised cell structure. The present study provides evidence that the reorganisation of physical state of water in germinating wheat seeds during hydration is essential for its subsequent event of germination. PMID- 12904912 TI - Microbial mats on the Orkney Islands revisited: microenvironment and microbial community composition. AB - The microenvironment and community composition of microbial mats developing on beaches in Scapa Flow (Orkney Islands) were investigated. Analysis of characteristic biomarkers (major fatty acids, hydrocarbons, alcohols, and alkenones) revealed the presence of different groups of bacteria and microalgae in mats from Waulkmill and Swanbister beach, including diatoms, Haptophyceae, cyanobacteria, and sulfate-reducing bacteria. These analyses also indicated the presence of methanogens, especially in Swanbister beach mats, and therefore a possible role of methanogenesis for the carbon cycle of these sediments. High amounts of algal lipids and slightly higher numbers (genera, abundances) of cyanobacteria were found in Waulkmill Bay mats. However, overall only a few genera and low numbers of unicellular and filamentous cyanobacteria were present in mats from Waulkmill and Swanbister beach, as deduced from CLSM (confocal laser scanning microscopy) analysis. Spectral scalar irradiance measurements with fiber optic microprobes indicated a pronounced heterogeneity concerning zonation and density of mainly anoxygenic phototrophs in Swanbister Bay mats. By microsensor and T-RFLP (terminal restriction fragment length polymorphism) analysis in Swanbister beach mats, the depth distribution of different populations of purple and sulfate-reducing bacteria could be related to the microenvironmental conditions. Oxygen, but also sulfide and other (inorganic and organic) sulfur compounds, seems to play an important role in the stratification and diversity of these two major bacterial groups involved in sulfur cycling in Swanbister beach mats. PMID- 12904913 TI - Dynamics of microbial populations and strong selection for Cycloclasticus pugetii following the Nakhodka oil spill. AB - Microbial population changes were monitored immediately after the Nakhodka oil spill accident in January 1997 at the heavily oil-contaminated Mikuni coast along the Sea of Japan. The total cell number was almost stable for one year at 2-5 x 10(5) cells mL(-1), while the relative occurrence of culturable heterotrophs and degraders of oil components such as C-heavy oil, kerosene, and n-tetradecane varied, showing a maximum (>50% of the total) immediately following the accident. Gene amplification and phylogenetic analysis of a dilution culture using C-heavy oil as the sole carbon and energy source revealed that one of the predominant oil degraders at the oil-contaminated coast in 2 weeks after the accident closely resembled the aromatic hydrocarbon decomposer Cycloclasticus pugetii. Microbial community composition in oil-contaminated seawater was estimated at the molecular level using newly developed oligonucleotide probes, probe wash-off curve estimation, and quantitative fluorescence dot-blot hybridization techniques. At two different oil-polluted sites, harbor and intertidal regions, the C. pugetii group was estimated to make up 23-25% of the total Bacteria population, followed by the aliphatic hydrocarbon decomposer Alcanivorax borkumensis, which formed 4 7% of the Bacteria. In incubation experiments using floated oil slick and indigenous microbes collected at the harbor, oil degradation activities were enhanced by the addition of both organic and inorganic nutrients. Significant decreases were found in aromatic and aliphatic hydrocarbon fractions: 54-60% and 22-24% in 2 weeks to 68-77% and 23-32% in 2 months, respectively. PMID- 12904915 TI - Temporal patterns in bacterial communities in three temperate lakes of different trophic status. AB - Despite considerable attention in recent years, the composition and dynamics of lake bacterial communities over annual time scales are poorly understood. This study used automated ribosomal intergenic spacer analysis (ARISA) to explore the patterns of change in lake bacterial communities in three temperate lakes over 2 consecutive years. The study lakes included a humic lake, an oligotrophic lake, and a eutrophic lake, and the epilimnetic bacterial communities were sampled every 2 weeks. The patterns of change in bacterial communities indicated that seasonal forces were important in structuring the behavior of the bacterial communities in each lake. All three lakes had relatively stable community composition in spring and fall, but summer changes were dramatic. Summertime variability was often characterized by recurrent drops in bacterial diversity. Specific ARISA fragments derived from these lakes were not constant among lakes or from year to year, and those fragments that did recur in lakes in different years did not exhibit the same seasonal pattern of recurrence. Nonetheless, seasonal patterns observed in 2000 were fairly successful predictors of the rate of change in bacterial communities and in the degree of autocorrelation of bacterial communities in 2001. Thus, seasonal forces may be important structuring elements of these systems as a whole even if they are uncoupled from the dynamics of the individual system components. PMID- 12904916 TI - Characterization of microbial diversity in hypersaline environments by melting profiles and reassociation kinetics in combination with terminal restriction fragment length polymorphism (T-RFLP). AB - The diversity of prokaryotes inhabiting solar saltern ponds was determined by thermal melting and reassociation of community DNA. These measurements were compared with fingerprinting techniques such as terminal restriction fragment length polymorphisms (T-RFLP) analysis, denaturant gradient gel electrophoresis (DGGE), and cloning and sequencing approaches. Three ponds with salinities of 22, 32, and 37% (NaCl saturation) were studied. The combination of independent molecular techniques to estimate the total genetic diversity provided a realistic assessment to reveal the microbial diversity in these environments. The changes in the prokaryotic communities at different salinity (22, 32, and 37% salt) were significant and revealed that the total genetic diversity increased from 22% to 32% salinity. At 37% salinity the diversity was reduced again to nearly half that at 22% salinity. Our results revealed that the community "genome" had a DNA complexity that was 7 (in 22% salinity pond), 13 (in 32% salinity pond), and 4 (in 37% salinity pond) times the complexity of an Escherichia coli genome. The base composition profiles showed two abundant populations, which changed in relative amount between the three ponds. They indicated an uneven taxon distribution at 22% and 37% salinity and a more even distribution at 32% salinity. The results indicated a large predominating population at 37% salinity, which might correspond to the abundance of square archaea (SPhT) observed by transmission electron microscopy (TEM) and also indicated by the same T-RFLP fragment as the SPhT. The SPhT phylotype has also been reported to be the most frequently retrieved phylotype from this environment by culture independent techniques. In addition, two different operational taxonomic units (OTU) were detected at 37% salinity based on PCR with bacterial specific primers and T-RFLP. One of these predominant phylotypes is the extreme halophilic bacterium belonging to the bacteroidetes group, Salinibacter ruber. PMID- 12904917 TI - Inter- and intrareader variability in the interpretation of two radiographic classification systems for juvenile rheumatoid arthritis. AB - OBJECTIVE: To evaluate the inter- and intrareader variability for interpretation of a modified Larsen's radiographic classification system for juvenile rheumatoid arthritis (JRA) focused on osteochondral lesions and a conventional Larsen's classification system, compared to a reference MR scoring system of corresponding images. MATERIALS AND METHODS: Seventy-five radiographs of 60 children with JRA, performed within a short interval of time from the MR examinations, were independently evaluated by three experienced radiologists, three diagnostic imaging residents and three rheumatologists, in two separate sessions, according to the two different classification methods, blinded to the corresponding MR images. RESULTS: The inter- and intrareader concordance rates between the two radiographic classification systems and the MR-related radiographs were respectively poor and poor/moderate. The interobserver range of weighted kappa values for the conventional and the modified Larsen's system respectively was 0.25-0.37 vs 0.19-0.39 for radiologists, 0.25-0.37 vs 0.18-0.30 for residents and 0.19-0.51 vs 0.17-0.29 for rheumatologists. The intrareader rate ranged from 0.17 0.55 for radiologists, 0.2-0.56 for residents, and 0.14-0.59 for rheumatologists. CONCLUSION: Although the proposal of a new radiographic classification system for JRA focused on osteochondral abnormalities sounds promising, the low inter- and intrareader concordance rates with an MR-related radiographic system makes the clinical applicability of such a radiographic system less suitable. PMID- 12904918 TI - Lenticulostriate echogenic vessels: clinical and sonographic study of 70 neonatal cases. AB - BACKGROUND: Neonatal branching echogenic streaks in the basal ganglia or 'lenticulostriate vasculopathy' (LSV) has no clear comprehensive aetiology. OBJECTIVE: To provide some clinical evidence, possibly relevant to aetiology, by analysis of a large series. MATERIALS AND METHODS: Seventy cases (9 deaths, 3 post mortem) identified between 1981 and 2000 out of 9,138 neonates with routine brain sonograms (578 in a protocol for neonates from HIV+ mothers). Review of maternal/neonatal charts for clinical data and serologic status. Retrospective analysis of sonograms for grading and time course of LSV and coexistent abnormalities. RESULTS: LSV was unilateral in 31 cases (22 right), bilateral in 39. Grading separated 8 major, 27 moderate and 35 minor patterns. In 42 cases, LSV was isolated, and in 28 it was mixed with peri-intraventricular haemorrhage grades I or II and/or leucomalacia. LSV was detected in the first postnatal week in 56 instances and appeared later in 13. Disappearance occurred mostly within 2 months. Perinatal circumstances: 13 recipient monochorionic twins (polycythaemia from twin-twin transfusion syndromes), 20 HIV+ mothers (possible immunological disturbances; only 2 infected neonates), 10 congenital anomalies, 1 toxoplasmosis, 1 postnatal cytomegalovirus infection, 25 miscellaneous conditions. Numerous maternal and neonatal factors were non-contributory. CONCLUSIONS: These results do not support congenital toxoplasmosis, rubella, cytomegalovirus, and herpes (TORCH) infections as the main causative factor. Polycythaemia and various immunological disturbances may be involved, as well as infectious agents not appropriately screened by routine serodiagnoses. PMID- 12904919 TI - Infants at higher risk to fracture than the general population of young infants. PMID- 12904920 TI - Balloon dilatation in children for oesophageal strictures other than those due to primary repair of oesophageal atresia, interposition or restrictive fundoplication. AB - BACKGROUND: Balloon dilatation of the oesophagus in children has been performed predominantly for treating strictures, which are the result of primary repair of oesophageal atresia, interposition surgery or restrictive Nissen's fundoplication. Reports of the use of this technique for alternative causes of stricture are few. OBJECTIVE: To report our experience and success with balloon dilatation of strictures due to caustic ingestion, achalasia, oesophagitis, congenital stenosis, and epidermolysis bullosa (EB) and to make comparisons with our treatment of patients with primary repair of oesophageal atresia (OA), as well as with reports in the English language literature. MATERIALS AND METHODS: Retrospective review of fluoroscopically guided balloon oesophageal dilatation procedures in 19 patients over a 5-year period, and comparison of those performed for OA repair complications with those due to other diseases. The average radiation dose, per procedure, was calculated by a medical physicist. RESULTS: Ten patients had strictures as a result of primary repair of OA. Three patients had stricture as a result of EB, two from achalasia, two from caustic injury, one due to an oesophageal web and one from reflux oesophagitis. Our results show that the technique can also be curative for the last group and that it may be used intermittently to alleviate symptoms in ongoing diseases. We have not experienced any complications and have also calculated that, even with prolonged use of multiple procedures, the radiation exposure is comparable to other radiological techniques. CONCLUSIONS: Patients with alternative causes for oesophageal stricture may be treated to resolution within 2 years using balloon dilatation. Ongoing diseases such as EB require ongoing dilatation, but balloon dilatation of strictures has been successful in alleviating swallowing difficulty. Patients with stricture from OA repair sometimes need ongoing dilatation. Radiation exposure for multiple procedures, over an extended period, is comparable to that from a single abdominal CT, and can be considered acceptable when repeat complex surgery is the alternative treatment option, or when ongoing incurable disease is the cause of the stricture. PMID- 12904921 TI - Commentary on "Inter- and intrareader variability in the interpretation of two radiographic classification systems for juvenile rheumatoid arthritis". PMID- 12904924 TI - Diffusion tensor imaging for long-term follow-up of corticospinal tract degeneration in amyotrophic lateral sclerosis. AB - Amyotrophic lateral sclerosis (ALS) is a predominantly clinical and electromyographic diagnosis. Conventional MRI reveals atrophy of the motor system, particularly the pyramidal tract, in the advanced stages but does not provide a sensitive measure of disease progression. Three patients with different principal symptoms of ALS, i.e., with predominant involvement of the upper (UMN) or lower (UMN) motor neurons, or bulbar disease, respectively, underwent serial clinical examination including lung function tests, conventional MRI, and diffusion tensor imaging (DTI). MRI demonstrated changes in of the pyramidal tract without measurable variation on follow-up. The patient with UMN involvement showed remarkable progressive loss of diffusion anisotropy in the pyramidal tract. DTI might be useful, together with clinical follow-up, as an objective morphological marker in therapeutic trials. PMID- 12904925 TI - Secondary aneurysmal bone cyst derived from a giant-cell tumour of the skull base. AB - We report a 20-year-old man with a giant-cell tumour of the skull base which showed multiple fluid levels on MRI. PMID- 12904926 TI - Concentric thalamic change on MR of acute necrotising encephalopathy of childhood. PMID- 12904927 TI - MRI of the tectorial and posterior atlanto-occipital membranes in the late stage of whiplash injury. AB - Our aim was to characterise and classify permanent structural changes in the tectorial and posterior atlanto-occipital membranes several years after a whiplash injury, and to evaluate the reliability of our classification. We obtained sagittal proton density-weighted images of the craniovertebral junction of 92 whiplash-injured and 30 uninjured individuals. Structural abnormalities in the two membranes were classified as grades 1-3 independently by three radiologists blinded for clinical information. Grading criteria were based on reduced tectorial membrane thickness, and elongation or rupture of the posterior atlanto-occipital membrane/dura mater complex. The same images were reassessed 4 months later. Image quality was graded good in 104 cases, slightly reduced in 13 and unsatisfactory in five. Of 117 tectorial membranes 31 (26.5%) showed grade 2 or 3 lesions, in the uninjured group none were grade 3 and only three were grade 2. Pair-wise interobserver agreement (weighted kappa) was moderate (0.47-0.50), while the intraobserver agreement was moderate to good (0.51-0.70). Of 117 posterior atlanto-occipital membranes 20 (17.1%) had grade 2 or 3 lesions; there was no grade 3 and only one grade 2 lesion in the uninjured group. Inter- and intraobserver agreement was good (0.61-0.74 and 0.65-0.86, respectively). Reduced image quality was the main reason for disagreement, but partial thinning and lateral tapering, as normal tectorial membrane variations, created difficulties in some cases. This study strongly indicates that whiplash trauma can damage the tectorial and posterior atlanto-occipital membranes; this can be shown on high resolution MRI. Better knowledge of normal anatomical variations and improved image quality should increase the reliability of lesion classification. PMID- 12904929 TI - Preoperative selective internal carotid artery dural branch embolisation for petroclival meningiomas. AB - We assessed the clinical value of preoperative embolisation of the dural branches of the internal carotid artery (ICA) in cases of petroclival meningioma was evaluated. We carried out preoperative selective embolisation on seven consecutive patients with large petroclival meningiomas, using nonbraided 2 F steam-shaped microcatheters and shapeable hydrophilic microguide-wires to enter the dural branches of the ICA. The embolisations were performed using digital subtraction fluoroscopy with 150-250 microm polyvinyl alcohol particles. The tumours were resected a few days after embolisation. We reviewed angiographic findings before and after endovascular devascularisation, blood loss during tumour resection and clinical outcome. All endovascular procedures were technically successful, and postembolisation angiography showed disappearance of or marked decrease in tumour stain in all cases. Intraoperative bleeding was easily controlled, and intraoperative blood loss was low. PMID- 12904928 TI - Contrast enhancement of the cochlear aqueduct in MR imaging: its frequency and clinical significance. AB - There have been no previous reports on contrast enhancement of the cochlear aqueduct in magnetic resonance imaging. The purpose of the present study was to evaluate the frequency and significance of this finding. Thirty-one patients (15 men and 16 women; age range 18-81 years) with otologic symptoms (sudden sensorineural hearing loss, vertigo, or tinnitus) were examined using contrast enhanced imaging on a 1.5-T MR scanner. The normal ear served as the control. Two radiologists evaluated contrast enhancement in the area of the cochlear aqueduct. Forty-eight of 62 ears (77.4%) showed contrast enhancement of the cochlear aqueduct, but no significant differences in the frequency of contrast enhancement were observed between patients with and patients without vertigo, tinnitus, sensorineural hearing loss, cerebellopontine angle tumors, or a high-riding jugular bulb. In addition, no gender- or age-related differences were noted. Contrast enhancement of the cochlear aqueduct was frequently observed, but the frequency of enhancement in symptomatic ears was not significantly higher than in control ears. The results of this study may prove helpful in avoiding unnecessary examinations and potential diagnostic confusion. PMID- 12904930 TI - Theophylline treatment may adversely affect the anoxia-induced erythropoietic response without suppressing erythropoietin production. AB - OBJECTIVE: To investigate the influence of theophylline on erythropoiesis in chronic obstructive pulmonary disease (COPD) and explore the potential underlying mechanisms. METHODS: We evaluated the haematological parameters and erythropoietin (EPO) values in 38 COPD patients, 18 of which had been treated with theophylline (8 mg/kg daily) for at least 1 year, and the other 20 had never received this drug; 38 sex- and age-matched healthy volunteers served as controls. We further studied the development of BFU-E (bursts forming units of erythrocyte precursors) -derived colonies in semisolid methylcellulose cultures in blood samples from 7 patients randomly selected from both groups. In addition, we studied the effects of theophylline on the erythroid cell development by adding this agent to erythroid cell cultures from 6 healthy volunteers at various concentrations. RESULTS: Haemoglobin values were found to be significantly lower in COPD patients treated with theophylline than in those untreated ( P<0.05). Both groups of patients exhibited significantly higher haemoglobin values than normal subjects ( P<0.01 and P<0.001 for treated and untreated patients, respectively). Serum EPO levels did not differ among the three studied groups. Unlike untreated patients and controls, the serum of the theophylline-treated patients produced a significant growth inhibition of erythroid bursts ( P<0.05); the in vitro use of theophylline showed a concentration-dependent inhibition ( P<0.001). CONCLUSION: Our findings confirm the decrease of red cell production, which occurs following administration of theophylline, exclude the possibility of decreased EPO synthesis and suggest a direct inhibitory action of theophylline on erythropoiesis. PMID- 12904931 TI - Inhibitory effect of 5-fluorouracil on human cytochrome P(450) isoforms in human liver microsomes. AB - OBJECTIVE: A number of case reports have been described regarding drug interactions with 5-fluorouracil (5-FU) and co-administered drugs. However, little is known regarding the inhibitory potential of 5-FU on the metabolism of co-administered drugs by cytochrome P(450) (CYP). The aim of the present study was to elucidate the inhibitory effect of 5-FU on CYP isoforms using human liver microsomes. METHODS: The inhibitory effect of 5-FU on CYP1A2, CYP2C9, CYP2C19, CYP2C8, CYP2E1, CYP2D6, and CYP3A4 activities was examined with specific probe drugs in human liver microsomes. RESULTS: 5-FU showed little or no inhibitory effect on CYP-catalyzed reactions in human liver microsomal preparations. CONCLUSION: 5-FU has no inhibitory effect on CYP isoforms or drug metabolism causing drug interaction with 5-FU. The mechanism that causes drug interaction between co-administered drugs and 5-FU may not be related to direct CYP inhibition by 5-FU. PMID- 12904932 TI - Foot-sole reflex receptive fields for human withdrawal reflexes in symmetrical standing position. AB - Human withdrawal-reflex receptive fields were assessed in 10 healthy subjects during standing with even support on both legs. Two electrical-stimulus intensities (1.2 and 2.2 times the pain threshold, PTh) were used. The painful stimuli were delivered in random order to 12 positions distributed over the foot sole. Tibialis anterior (TA), soleus (SO), vastus lateralis (VL), semitendinosus (ST), and iliopsoas (IL) reflexes were recorded. Further, the vertical force was recorded and the center or pressure (CoP) was assessed in the frontal and sagittal planes on both legs. Reflexes were observed at both intensities with the strongest reflexes at the high intensity. Around the ankle joint, SO reflexes dominated, which is in contrast to previous observations for subjects sitting. An unloading of the limb was found on the stimulated leg associated with a simultaneous loading of the contralateral leg. The shift in load was most pronounced for stimulation of the heel. The flexors ST and IL also had strong reflexes with reflex patterns correlated to the pattern of unloading. The shift in vertical force was accomplished by a move of the CoP in the anterior direction on the stimulated limb (contraction of SO), which simultaneously caused a small movement of the CoP in the lateral direction. In the present standing conditions, the ankle extensor played a dominant role in the withdrawal pattern in contrast to previous studies during sitting, relaxed conditions. PMID- 12904933 TI - Quantitative analyses of principal and secondary compound parieto-occipital feedback pathways in cat. AB - The purpose of our study was to quantify the magnitude of principal and secondary pathways emanating from the middle suprasylvian (MS) region of visuoparietal cortex and terminating in area 18 of primary visual cortex. These pathways transmit feedback signals from visuoparietal cortex to primary visual cortex. (1) WGA-HRP was injected into area 18 to identify inputs from visual structures. In terms of numbers of neurons, feedback projections to area 18 from MS sulcal cortex (areas PMLS, AMLS and PLLS) comprise 26% of inputs from all visual structures. Of these neurons, between 21% and 34.9% are located in upper layers 2 4 and the dominant numbers are located in deep layers 5 and 6. Areas 17 (11.8%) and 19 (11.2%) provide more modest cortical inputs, and another eight areas provide a combined total of 4.3% of inputs. The sum of neurons in all subcompartments of the lateral geniculate nucleus (LGN) accounts for another 34.8% of the input to area 18, whereas inputs from the lateral division of the lateral-posterior nucleus (LPl) account for the final 11.9%. (2) Injection of tritiated-((3)H)-amino acids into MS sulcal cortex revealed substantial direct projections from MS cortex that terminated in all layers of area 18, but with a markedly lower density in layer 4. Projections from MS cortex to both areas 17 and 19 are of similar density and characteristics, whereas those to other cortical targets have very low densities. Quantification also revealed minor-to modest axon projections to all components of LGN and a massive projection throughout the LP-Pul complex. (3) Superposition of the labeled terminal and cell fields identified secondary, compound feedback pathways from MS cortex to area 18. The largest secondary pathway is massive and it includes the LPl nucleus. Much more modest secondary pathways include areas 17 and 19, and LGN. The relative magnitudes of the secondary pathways suggest that the one through LPl exerts a major influence on area 18, whereas the others exert more modest or minor influences. MS cortex in the contralateral hemisphere also innervates area 18 directly. These data are important for interpreting the impact of deactivating feedback projections from visuoparietal cortex on occipital cortex. PMID- 12904935 TI - Humans use continuous visual feedback from the hand to control fast reaching movements. AB - How visual feedback contributes to the on-line control of fast reaching movements is still a matter of considerable debate. Whether feedback is used continuously throughout movements or only in the "slow" end-phases of movements remains an open question. In order to resolve this question, we applied a perturbation technique to measure the influence of visual feedback from the hand at different times during reaching movements. Subjects reached to touch targets in a virtual 3D space, with visual feedback provided by a small virtual sphere that moved with a subject's fingertip. Small random perturbations were applied to the position of the virtual fingertip at two different points in the movement, either at 25% or 50% of the total movement extent. Despite the fact that subjects were unaware of the perturbations, their hand trajectories showed smooth and accurate corrections. Detectable responses were observed within an average of 160 ms after perturbations, and as early as 60% of the distance to the target. Response latencies were constant across different perturbation times and movement speed conditions, suggesting that a fixed sensori-motor delay is the limiting factor. The results provide direct evidence that the human brain uses visual feedback from the hand in a continuous fashion to guide fast reaching movements throughout their extent. PMID- 12904934 TI - Muscle synergies during shifts of the center of pressure by standing persons. AB - Movements by a standing person are commonly associated with adjustments in the activity of postural muscles to cause a desired shift of the center of pressure (COP) and keep balance. We hypothesize that such COP shifts are controlled (stabilized) using a small set of central variables (muscle modes, M-modes), while each M-mode induces changes in the activity of a subgroup of postural muscles. The main purpose of this study has been to explore the possibility of identification of muscle synergies in a postural task using the framework of the uncontrolled manifold (UCM) hypothesis employing the following three steps in data analysis: (i) Identification of M-modes: Subjects were asked to release a load from extended arms through a pulley system, resulting in a COP shift forward prior to load release. Electromyographic (EMG) activity of eleven postural muscles on one side of the body was integrated over a 100 ms interval corresponding to the early stage of the COP shift, and subjected to a principal component (PC) analysis across multiple repetitions of each task. Three PCs were identified and associated with a 'push-back M-mode', a 'push-forward M-mode' and a 'mixed M-mode'. (ii) Calculation of the Jacobian of the system, which relates changes in the magnitude of M-modes to COP shifts using regression techniques: Subjects performed three different tasks (releasing different loads at the back, voluntarily shifting body weight forward and backward, at different speeds) to verify if the relationship between magnitudes of M-modes and COP shifts is task or direction specific. (iii) UCM analysis: Three tasks were chosen (load release in the front, arm movement forward and backward) which were associated with an early shift in COP. A manifold was identified in the M-mode space corresponding to a certain average (across trials) shift of the COP and variance per degree of freedom within the UCM (V(UCM)) and orthogonal (V(ORT)) to the UCM was computed. Across subjects, V(UCM) was significantly higher than V(ORT) when analysis at the third step was performed using a Jacobian computed based on a set of tasks associated with a COP shift in the same direction but not in the opposite direction. This result confirms our hypothesis that the M-modes work together as a synergy to stabilize a desired shift of the COP. Forward and backward COP shifts are associated with different synergies based on the same three M-modes. PMID- 12904936 TI - Locomotor adjustments for circumvention of an obstacle in the travel path. AB - Independent living requires the navigation of a surrounding environment which is often cluttered with obstacles. When walking around an obstacle in the travel path, safe clearance requires some degree of body-segment reorientation. While body-segmental coordination strategies have been well studied for steering tasks that require moving the body in a new walking direction, it has never been established just what coordination strategies are used in different walking tasks. To address this issue, the current study was designed to investigate the timing of body segment coordination strategies and whole-body anticipatory locomotor adjustments employed when circumventing an obstacle placed in the travel path. Six healthy adults were asked to walk at their natural pace during unobstructed walking, as well as during avoidance to the right or left of a cylindrical obstacle (OBS) located in the travel path. Data analyzed were center of mass (COM) clearance from the OBS, forward velocity, step length and width, yaw angles of the head and trunk, roll angle of the trunk, and medial-lateral COM displacement. Onset of change in these variables from unobstructed walking was calculated as the time from OBS crossing. Avoidance involved two equally used strategies: lead limb close to or away from the OBS during the crossing step. Medial-lateral COM deviations were controlled by changes in step width without changes in trunk roll. There were no differences in the onset times of body segment reorientation for path deviation. These results are in contrast to previous studies on change in travel direction where the head segment initiates the body reorientation. Contrary to a steering task, circumventing an obstacle requires a different coordination for a transient change in COM trajectory with the underlying travel-direction maintained. PMID- 12904937 TI - Startle response is dishabituated during a reaction time task. AB - Recent experiments pairing a startling stimulus with a simple reaction time (RT) task have shown that when participants are startled, a prepared movement may be triggered earlier in comparison to voluntary initiation (Carlsen et al. 2003, in press; Siegmund et al. 2001; Valls-Sole et al. 1999). The use of this paradigm in experiments may provide new insights into processes that control rapid voluntary actions. However, because the startle response habituates with repeated exposure to the startling stimulus, its use in experiments may be limited. Previously Brown et al. (1991) and later Siegmund et al. (2001) noted that individuals habituate to a startling stimulus at different rates depending on the required activity level of the participant in the task. The present experiment was designed to determine the rate at which participants habituate to a startle during the completion of a RT task. Participants completed 100 trials in which an active wrist extension to a target was performed as fast as possible following an auditory tone. An unexpected 124 dB auditory startle stimulus accompanied the imperative stimulus in 20 of these trials. For the duration of the experiment, startle response electromyographic (EMG) activity continued to be produced in the sternocleidomastoid muscle (SCM) indicating that habituation was not complete after 20 startle trials. Furthermore RT in the startle condition was significantly shorter than control RT. However, findings indicate that when a measurable EMG burst in the SCM was present, RT was significantly shorter than when no SCM burst was present. PMID- 12904938 TI - Lubeluzole and conditioned learning after cerebral ischemia. AB - Lubeluzole [S-4-(2-benzothiazolylmethylamino)-alpha-((3,4-difluorophenoxy)methyl) 1-piperidineethanol] reduces the severity of cerebral injury in animal models of brain ischemia. Its beneficial effects may include decreased concentration of extracellular glutamate, blockade of sodium and calcium channels, and attenuation of nitric oxide-mediated neuronal death. Previous studies have shown that global cerebral ischemia in rabbits impaired the subsequent acquisition of a trace conditioned eyeblink reflex. Here, we examined the effect of preischemic treatment with lubeluzole on the acquisition of a trace-conditioned eyeblink response after 6.5 min of global cerebral ischemia. Three groups of rabbits underwent cerebral ischemia: one group underwent ischemia alone (I) and two groups underwent ischemia and also received lubeluzole (L(1), 1.25 mg/kg, and L(2), 2.5 mg/kg). All animals were subsequently trained using classical trace conditioning. Each training session consisted of the presentation of the conditioned stimulus (an 85-dB, 6-kHz auditory tone lasting for 100 ms) followed by a trace interval (a period of 300 ms during which no external stimulus was delivered) followed finally by the delivery of the unconditioned stimulus (a 150 ms puff of air directed at the cornea). We found that animals receiving preischemic administration of 1.25 mg/kg of lubeluzole demonstrated a significantly improved acquisition of the trace-conditioned reflex as compared to animals that did not receive lubeluzole. This finding demonstrates improved long term neurobehavioral outcome with preischemic administration of 1.25 mg/kg of lubeluzole. PMID- 12904939 TI - Effects of motivational conflicts on visually elicited saccades in monkeys. AB - The prospect of reward evoked by external stimuli is a central element of goal oriented behavior. To elucidate behavioral effects of reward expectation on saccade latency, we employed a visually guided saccade task with asymmetrical reward schedule. The monkey had to make an immediate saccade to a peripheral visual target in every trial, but was rewarded for a correct saccade to only one of four possible target positions. Reward availability was predictable on the basis of the spatial position of the target throughout a daily session. Compared with the condition where all positions were rewarded with a smaller amount, the mean saccade latency in the asymmetrical reward schedule was significantly shorter when the saccade was made toward the position associated with reward than when it was directed to no-reward positions. Furthermore, a divergence-point analysis on cumulative latency distributions showed that the expectation of reward facilitated saccades at all latency ranges. In contrast, the expected lack of reward delayed the initiation of saccades with latencies longer than about 200 ms, irrespective of whether the saccade was made to a position orthogonal or opposite to the reward position. For saccades with latencies of more than approximately 240 ms, an additional delay was observed when the saccade was made to a position opposite, as compared to orthogonal, to the reward position. These results suggest that the facilitation by predictive reward is mediated by a preparatory process that is location-specific, whereas the inhibition by the absence of reward takes about 200 ms after the target onset to become effective and is initially location nonspecific but turns location-specific over time. PMID- 12904940 TI - Neuromuscular and biomechanical coupling in human cycling: adaptations to changes in crank length. AB - This study exploited the alterations in pedal speed and joints kinematics elicited by changing crank length (CL) to test how altered task mechanics during cycling will modulate the muscle activation characteristics in human rectus femoris (RF), biceps femoris long head (BF), soleus (SOL) and tibialis anterior (TA). Kinetic (torque), kinematic (joint angle) and muscle activity (EMG) data were recorded simultaneously from both legs of 10 healthy adults (aged 20-38 years) during steady-state cycling at ~60 rpm and 90-100 W with three symmetrical CLs (155 mm, 175 mm and 195 mm). The CL elongation (DeltaCL) resulted in similar increases in the knee joint angles and angular velocities during extension and flexion, whilst the ankle joint kinematics was significantly influenced only during extension. DeltaCL resulted in significantly reduced amplitude and prolonged duration of BF EMG, increased mean SOL and TA EMG amplitudes, and shortened SOL activity time. RF activation parameters and TA activity duration were not significantly affected by DeltaCL. Thus total SOL and RF EMG activities were similar with different CLs, presumably enabling steady power output during extension. Higher pedal speeds demand an increased total TA EMG activity and decreased total BF activity to propel the leg through flexion into extension with a greater degree of control over joint stability. We concluded that the proprioceptive information about the changes in the cycling kinematics is used by central neural structures to adapt the activation parameters of the individual muscles to the kinetic demands of the ongoing movement, depending on their biomechanical function. PMID- 12904942 TI - Differential analysis of phosphorylated proteins in resting and thrombin stimulated human platelets. AB - Blood platelets are important components of haemostasis. After their activation they cause healing of wounds by forming plugs and initiate repair processes. One important event in regulating this activation is the phosphorylation/dephosphorylation of multiple proteins on various tyrosine, serine and threonine residues. To understand the exact molecular mechanisms in platelet activation it is essential to identify proteins involved in the signalling pathways and to localise and characterise their phosphorylation sites. After treatment with (32)P and separation by 2D-PAGE using different pI ranges, phosphorylated platelet proteins were detected by autoradiography. Phosphotyrosine-containing proteins were assigned by immunoblotting with an anti phosphotyrosine antibody. Another approach for the identification of phosphorylated proteins was immunoprecipitation of tyrosine-phosphorylated proteins using an anti-phosphotyrosine antibody. Protein spots/bands of interest were excised from the gel, digested with trypsin and analysed by MALDI-TOF-MS and nano-LC-ESI-MS/MS, respectively. Several phosphorylated proteins could be identified and the localisation of some in vivo phosphorylation sites was possible. PMID- 12904941 TI - Static and dynamic changes in body orientation modulate spinal reflex excitability in humans. AB - In the present study, we investigated the modulation pattern of the soleus H reflex in healthy subjects in response to imposed static and dynamic changes in body angle, referenced to the vertical plane. Soleus H reflexes were recorded using conventional methods with subjects either supine or while they were erect. Changes in body angle were initiated with subjects lying supine on a tilt table. Table position was controlled via a motor and could move from the horizontal to the upright position and beyond. Elastic bands around the trunk (upper and lower part) and around the thigh and shank secured subjects' position. In the vertical position, the soleus H reflex exhibited a strong depression in all subjects tested, reaching amplitudes as low as 40+/-8.1% of the control reflex (Ho). With subjects supine, positioning the body at 10 degrees, 20 degrees, 40 degrees, 60 degrees, 90 degrees, -50 degrees and -20 degrees all resulted in a significant facilitation of the soleus H reflex. The reflex magnitude at these angles ranged from 140+/-17.2% to 180+/-10.9% of the Ho. Reflex facilitation was also observed following dynamic tilt of the body in the sagittal plane (at 1.8 degrees /s) with the H reflex reaching amplitudes as high as 300+/-18.3% of Ho. Our findings indicate that changes in body orientation induced a significant facilitation of the H reflex magnitude in soleus motoneurones that were essentially independent of angular change in body orientation or of movement direction. In addition, they highlight the potent modulatory effects that natural stimulation of the vestibular system can have on reflex excitability. The implications of our findings are discussed in relation to the maintenance of body posture. PMID- 12904944 TI - High-performance liquid chromatography-fluorescent assay of catechol-O methyltransferase activity in rat brain. AB - A method to measure catechol- O-methyltransferase (COMT) activity using high performance liquid chromatography-fluorescence detection with norepinephrine (NE) as a natural substrate was optimized for both soluble (S-) and membrane-bound (MB ) COMT activities in rat brain areas, cerebral cortex, cerebellum, hippocampus, brain stem, hypophysis, and hypothalamus. The highest S-COMT activity in Sprague Dawley rat brain was found in hippocampus. MB-COMT activities in all brain areas were about 3-8 times lower than S-COMT activities. However, considering Vmax/ Km values, specificity constants for NE to S- and MB-COMT contributes mainly to the metabolism of NE in cerebral cortex and cerebellum. PMID- 12904943 TI - Acetylation of the HIV-1 Tat protein: an in vitro study. AB - In the last few years, the understanding of lysine acetylation as a regulatory post-translational modification of proteins in cell signalling cascades has increased. It is now known that not only histones but also non-histone factors can serve as substrates of different acetyltransferase enzymes. Acetylated lysine residues in non-histone factors are often identified using radioactive labelling experiments and immunochemical analysis of synthetic peptides. In this study of the human immunodeficiency virus 1 (HIV-1) Tat protein, we demonstrate the benefits of matrix-assisted laser desorption/ionisation mass spectrometry, proteolytic digestion and Edman sequencing for the mapping of acetylation sites. We confirmed that the HIV-1 Tat protein is acetylated in vitro by the acetyltransferase p300 at a specific lysine residue at position 50 in its RNA binding region. Furthermore, we showed that the Tat cysteine-rich region is acetylated at multiple cysteine residues in the absence of enzyme. Since this non enzymatic cysteine acetylation occurs independently from the surrounding peptide sequence, we consider the presence of cysteine residues in acetylated peptides an important factor for the interpretation of in vitro acetylation assays in general. PMID- 12904945 TI - Analysis of alcohol ethoxylates and alkylamine ethoxylates in agricultural soils using pressurised liquid extraction and liquid chromatography-mass spectrometry. AB - Nonionic surfactants e.g. alcohol ethoxylates (AEOs) and alkylamine ethoxylates (ANEOs) are commonly utilised as adjuvants in pesticide formulations to enhance their effectiveness. In this study, analytical methods for AEO and ANEO determination in soil samples using pressurised liquid extraction (PLE) were developed and used in connection with LC-MS. The recovery of the method, which was highly dependent on the soil properties, varied in the range 47-106% for AEO and 27-109% for ANEO. Detection limits (LOD) were 7-13 microg kg(-1) for AEO and 24-43 microg kg(-1) for ANEO. The developed method has been applied to determine AEOs and ANEOs in surface soil samples from fields sprayed with glyphosate herbicides. Tallowalkylamine ethoxylates (an ANEO) were detected in the soil before and after pesticide application, with increasing concentrations after treatment. The highest concentration in the soil samples was observed for the ANEO homologues with the longest ethoxy chains; in the clay soil the concentration decreased with the length of the ethoxy chain. ANEOs added to pesticide formulations as a technical mixture will, as demonstrated in this study, behave as individual homologues, which is reflected in their behaviour in the environment. PMID- 12904946 TI - Label-free screening of bio-molecular interactions. AB - The majority of techniques currently employed to interrogate a biomolecular interaction require some type of radio- or enzymatic- or fluorescent-labelling to report the binding event. However, there is an increasing awareness of novel techniques that do not require labelling of the ligand or the receptor, and that allow virtually any complex to be screened with minimal assay development. This review focuses on three major label-free screening platforms: surface plasmon resonance biosensors, acoustic biosensors, and calorimetric biosensors. Scientists in both academia and industry are using biosensors in areas that encompass almost all areas drug discovery, diagnostics, and the life sciences. The capabilities and advantages of each technique are compared and key applications involving small molecules, proteins, oligonucleotides, bacteriophage, viruses, bacteria, and cells are reviewed. The role of the interface between the biosensor surface (in the case of SPR and acoustic biosensors) and the chemical or biological systems to be studied is also covered with attention to the covalent and non-covalent coupling chemistries commonly employed. PMID- 12904947 TI - Use of solid-phase extraction to eliminate interferences in the determination of mercury by flow-injection CV AAS. AB - Solid-phase extraction with two-step elution has been developed for effective elimination of copper and iron interference with mercury determination by flow injection cold vapour atomic absorption spectrometry (CV AAS). Sodium tetrahydroborate(III) was used as reducing agent. Cation-exchanger Dowex 50Wx4 was applied for the sorption of mercury and both interfering ions. In the first step elution of Cu(II) and Fe(III) was performed using 0.5 mol L(-1) KF solution. Then mercury was eluted with 0.1% thiourea in 8% HCl. The detection limit (3 delta) for Hg(II) was 27 ng L(-1). The expanded uncertainty estimated for the whole procedure was about 6%. The accuracy of the proposed method was evaluated by determination of the recovery of known amount of mercury added to mineral, spring, and tap waters, and by analysis of a certified reference material BCR 144R (sewage sludge). PMID- 12904948 TI - Fast LC/MS in the analysis of small molecules. AB - Liquid chromatography-mass spectrometry (LC/MS) has become one of the most widely used analytical techniques in both qualitative and quantitative analysis of small molecules. Recently, with the increasing demand for ever-higher sample throughput, the use of faster chromatographic separations has become popular, along with other LC/MS methods that decrease analytical cycle-time. The burgeoning use of LC/MS has meant that the primary expertise of many practitioners today is not in the field of LC/MS, which has been facilitated by the ease-of-use of modern LC/MS systems. An examination of the current state of the literature, relating to "fast LC/MS", should serve well to those new to LC/MS, and should help them in the development of fast LC/MS methods that are effective in terms of both the chromatography and the utilization of the mass spectrometer. This review paper focuses on fast LC/MS analyses of small molecules that have been reported in peer-reviewed publications. PMID- 12904949 TI - "A focus on bioanalytical chemistry in Spain" from papers presented to: "Jornadas de Analisis Instrumental (JAI)" 26-29 November 2002, Barcelona, Spain. PMID- 12904950 TI - Effect-directed analysis: a promising tool for the identification of organic toxicants in complex mixtures? AB - Wastewater effluents, groundwater, surface water, sediments, soils and air particulate matter are often contaminated by a multitude of chemicals. Since often no a priori knowledge of relevant toxicants exists, chemical analysis alone is not an appropriate tool for hazard assessment. Instead, a linkage of effect data and hazardous compounds is required. For that purpose, effect-directed analysis (EDA) was developed, which is based on a combination of biotesting, fractionation procedures and chemical analytical methods. Since a controversial discussion about the prospects of success in relation to the expense exists, the current methodological state of EDA for organic toxicants in complex mixtures and important results are reviewed in this paper with the aim of establishing criteria for the successful use of this promising tool. While EDA is a powerful tool to identify specifically acting individual toxicants close to the source of emission, it is inappropriate for screening purposes and often may fail in remote areas where the concentrations of specific toxicants are too low relative to the nonspecific toxicity of the whole mixture of natural and anthropogenic compounds. The biological tools have to be carefully selected with respect to their ability to detect specific effects and their significance in hazard assessment. Sophisticated chemical tools are required to identify individual toxicants in mixtures of thousands of compounds, which are typical for contaminated environments. PMID- 12904951 TI - Quantitative determination of saturated oligogalacturonic acids in enzymatic digests of polygalacturonic acid, pectin and carrot pomace by on-line LC-ESI-MS. AB - A method for the quantitative determination of oligogalacturonic acids (OGAs) by on-line high-performance liquid chromatographic (HPLC) separation and mass spectrometric detection via an electrospray interface (ESI-MS) without additional desalting steps was developed. Saturated OGAs up to a degree of polymerization (dp) of 3 were quantified by comparison with reference compounds. The calibration plots showed high linearity (R(2)>0.99), and the detection limits for dp 1, 2, and 3 were 11, 28, and 6 ng per injection, respectively. Non- and partially methyl-esterified OGAs with a dp of 3 and 4 were calculated semi-quantitatively as dp 3. The analytical system was used for the quantification of OGAs of digests obtained by incubation of polygalacturonic acid, pectin, and carrot pomace with commercial enzyme preparations. Furthermore, methyl-esterified OGAs up to a dp of 12 containing up to 4 methyl esters were detected in a pectin digest. PMID- 12904952 TI - Analysis of meso-2,3-dimercaptosuccinic acid in human urine by capillary electrophoresis using direct injection. AB - meso-2,3-Dimercaptosuccinic acid ( meso-DMSA) is an effective chelating agent for the treatment of lead poisoning. We have developed a capillary electrophoresis (CE) method to monitor the urinary excretion of meso-DMSA in human beings. The urine sample was directly injected for analysis in CE without the requirement of solid-phase extraction (SPE). The meso-DMSA was detected in 20 mM borate buffer (pH 8.3) using a 60-cm length bare fused-silica capillary (75-microm ID, 52.5-cm effective length). The meso-DMSA can be extensively biotransformed during metabolism, and no meso-DMSA in urine samples was found in our studies. Any metabolized meso-DMSA can be successfully converted to free meso-DMSA by chemical reduction with dithiothreitol (DTT). In addition, samples were also treated with ethylenediaminetetraacetic acid (EDTA) to transchelate any meso-DMSA that is coordinated with metal ions present in the urine samples. The total amount of meso-DMSA present as these chemical forms was quantified after chemical reduction and addition of EDTA. The detection limit of meso-DMSA was about 50 microM, the RSD of peak area and migration time of meso-DMSA were 4-8% and less than 1%, respectively. PMID- 12904953 TI - Biosensors based on enzyme field-effect transistors for determination of some substrates and inhibitors. AB - This paper is a review of the authors' publications concerning the development of biosensors based on enzyme field-effect transistors (ENFETs) for direct substrates or inhibitors analysis. Such biosensors were designed by using immobilised enzymes and ion-selective field-effect transistors (ISFETs). Highly specific, sensitive, simple, fast and cheap determination of different substances renders them as promising tools in medicine, biotechnology, environmental control, agriculture and the food industry. The biosensors based on ENFETs and direct enzyme analysis for determination of concentrations of different substrates (glucose, urea, penicillin, formaldehyde, creatinine, etc.) have been developed and their laboratory prototypes were fabricated. Improvement of the analytical characteristics of such biosensors may be achieved by using a differential mode of measurement, working solutions with different buffer concentrations and specific agents, negatively or positively charged additional membranes, or genetically modified enzymes. These approaches allow one to decrease the effect of the buffer capacity influence on the sensor response in an aim to increase the sensitivity of the biosensors and to extend their dynamic ranges. Biosensors for the determination of concentrations of different toxic substances (organophosphorous pesticides, heavy metal ions, hypochlorite, glycoalkaloids, etc.) were designed on the basis of reversible and/or irreversible enzyme inhibition effect(s). The conception of an enzymatic multibiosensor for the determination of different toxic substances based on the enzyme inhibition effect is also described. We will discuss the respective advantages and disadvantages of biosensors based on the ENFETs developed and also demonstrate their practical application. PMID- 12904954 TI - Development of analytical procedures for determination of total chromium by quadrupole ICP-MS and high-resolution ICP-MS, and hexavalent chromium by HPLC-ICP MS, in different materials used in the automotive industry. AB - A European directive was recently adopted limiting the use of hazardous substances such as Pb, Hg, Cd, and Cr(VI) in vehicle manufacturing. From July 2003 a maximum of 2 g Cr(VI) will be authorised per vehicle in corrosion preventing coatings of key components. As no standardised procedures are available to check if produced vehicles are in agreement with this directive, the objective of this work was to develop analytical procedures for total chromium and Cr(VI) determination in these materials. The first step of this study was to optimise digestion procedures for total chromium determination in plastic and metallic materials by inductively coupled plasma mass spectrometry (ICP-MS). High resolution (HR) ICP-MS was used to examine the influence of polyatomic interferences on the detection of the (52)Cr(+) and (53)Cr(+) isotopes. If there was strong interference with m/ z 52 for plastic materials, it was possible to use quadrupole ICP-MS for m/ z 53 if digestions were performed with HNO(3)+H(2)O(2). This mixture was also necessary for digestion of chromium from metallic materials. Extraction procedures in alkaline medium (NH(4)(+)/NH(3) buffer solution at pH 8.9) assisted by sonication were developed for determining Cr(VI) in four different corrosion-preventing coatings by HPLC-ICP-MS. After optimisation and validation with the only solid reference material certified for its Cr(VI) content (BCR 545; welding dusts), the efficiency of this extraction procedure for screw coatings was compared with that described in the EN ISO 3613 standard generally used in routine laboratories. For coatings comprising zinc and aluminium passivated in depth with chromium oxides the extraction procedure developed herein enabled determination of higher Cr(VI) concentrations. This was also observed for the screw covered with a chromium passivant layer on zinc nickel. For coating comprising a chromium passivant layer on alkaline zinc the standardized extraction procedure was more efficient. In the case of painted metallic plate, because of a reactive matrix towards Cr(VI), its extraction without degradation was difficult to perform. PMID- 12904955 TI - Determination of alcohol content in beverages using short-wave near-infrared spectroscopy and temperature correction by transfer calibration procedures. AB - This paper reports the utilization of short-wave near-infrared (SW-NIR) transmission spectroscopy for rapid and conclusive analysis of alcoholic content (% v/v) in beverages. This spectral region is interesting because common visible diode array spectrometers can be utilized, reducing time and costs in comparison with traditional near-infrared or mid-infrared instruments. A correction of external temperature influence is necessary, and for this purposes two calibration transfer procedures were compared: piecewise direct standardization (PDS) and orthogonal signal correction (OSC). The RMSEP found for the alcoholic content model at 20 degrees C was 0.13% v/v and, after application of transfer calibration procedures at other temperatures (15, 25, 30 and 35 degrees C) using the model built at 20 degrees C, errors of the same order of magnitude were obtained. PMID- 12904956 TI - A flow-injection ultrafiltration sampling chemiluminescence system for on-line determination of drug-protein interaction. AB - A flow-injection ultrafiltration sampling chemiluminescence system for on-line determination of cimetidine-bovine serum albumin (BSA) interaction is proposed in this paper. Cimetidine can be oxidized by N-bromosuccinimide (NBS) and sensitized by fluorescein to produce high chemiluminescence emission in basic media. The concentration of cimetidine is linear with the CL intensity in the range 3 x 10( 7) - 1 x 10(-4) mol L(-1) with a detection limit of 1 x 10(-7) mol L(-1) (3 sigma). The drug and protein were mixed in different molar ratios in 0.067 mol L( 1) phosphate buffer, pH 7.4, and incubated at 37 degrees C in a water bath. The ultrafiltration probe was utilized to sample the mixed solution at a flow rate of 5 micro L min(-1). The data obtained by the proposed ultrafiltration flow injection chemiluminescence method was analyzed with Scrathard analysis and a Klotz plot. The estimated association constant ( K) and the number of the binding site ( n) on one molecule of BSA by Scrathard analysis and Klotz plot were 3.15 x 10(4) L mol(-1) and 0.95, 3.25 x 10(4) L mol(-1) and 0.92, respectively. The proposed system proved that flow-injection chemiluminescence analysis coupled with on-line ultrafiltration sampling is a simple and reliable technique for the study of drug-protein interaction. PMID- 12904957 TI - The use of gold bands for flow immunoelectrochemical devices. AB - Gold bands sputtered over a polymeric material, Kapton, have been employed not only for electrochemical detection but also for the development of enzyme immunoassays in a flow system. The immunological interactions on bands acting as reactors are considered for a model analyte, IgM. Different formats of flow immunoassays, competitive and non-competitive, have been checked. Compared with previous results, automation gives rise to in a reduction in analysis time and in reagent consumption. Lower limits of detection are also obtained. Detection, which is also carried out in the flow system, is based on the oxidation of naphthol, the product of the enzymatic hydrolysis of naphthyl-phosphate. PMID- 12904958 TI - 2,3-Dihydroxypyridine-loaded cellulose: a new macromolecular chelator for metal enrichment prior to their determination by atomic absorption spectrometry. AB - New macromolecular chelators have been synthesized, by loading 2,3 dihydroxypyridine (DHP) on cellulose via linkers -NH-CH(2)-CH(2)-NH-SO(2) C(6)H(4)-N=N- and -SO(2)-C(6)H(4)-N=N-, and characterized by elemental analysis, TGA, IR, and CPMAS (13)C NMR spectra. The cellulose with DHP anchored by the shorter linker had better sorption capacity (between 69.7 and 431.1 micromol g( 1)) for Co(II), Ni(II), Cu(II), Zn(II), Cd(II), Pb(II), and Fe(III)) than the other (51.9-378.1 micromol g(-1)); the former was therefore studied in detail as a solid extractant for these metal ions. The optimum pH ranges for quantitative sorption (recovery 97.6-99.8%) on this matrix were: 7.0-9.0, 6.0-9.0, 3.0-8.0, 6.0-8.0, 6.0-9.0, 6.0-7.0, and 2.0-6.0 respectively. Desorption was quantitative with 0.5 mol L(-1) HCl and 0.5 mol L(-1) HNO(3) (for Pb). Simultaneous sorption (at pH 7.0) of all metal ions other than Fe(III) was possible if their total concentration did not exceed the sorption capacity (lowest value). The recovery of seven metal ions from their mixture at pH 6.0 was nearly quantitative when the concentration level of each metal ion was 0.2 microg mL(-1). The optimum flow rate of metal ion solutions for quantitative sorption of metal onto a column packed with DHP-modified cellulose was 2-7 mL min(-1), whereas for desorption the optimum flow rate for the acid solution was 2-4 mL min(-1). The time needed to reach 50% of the total loading capacity ( t(1/2)) was <5 min for all the metal ions except Ni and Pb. The limit of detection (blank+3 s) was from 0.70 to 4.75 microg L(-1) and the limit of quantification (blank+10 s) was between 0.79 and 4.86 microg L(-1). The tolerance limits for NaCl, NaBr, NaI, NaNO(3), Na(2)SO(4), Na(3)PO(4,) humic acid, EDTA, Ca(II), and Mg(II) for sorption of all metal ions are reported. The column packed with DHP-anchored cellulose can be reused at least 20 times for enrichment of metal ions in water sample. It has been used to enrich all the metal ions in pharmaceutical and water samples before their determination by flame AAS. RSD for these determinations was between 1.1 and 6.9%. PMID- 12904959 TI - Derivatisation in gas chromatographic determination of acidic herbicides in aqueous environmental samples. AB - This paper reviews derivatisation processes applied in chromatographic determination of acidic herbicides (with carboxyl and phenol groups), mainly in aqueous environmental samples. The discussion focuses on the basic derivatisation reactions used to convert herbicides to derivatives to make them analysable by gas chromatography, and possibly to reduce detection limits and/or increase extraction recovery from aqueous samples. The reactions are transesterification, esterification, silylation, alkylation, and extractive and pyrolytic alkylation. The reagents used to conduct the reactions are numerous. Diazomethane is a very efficient methylation reagent but explosive and toxic. Methyl iodide also ensures rapid and efficient methylation. Benzyl bromide can be used directly in water but derivatisation yield is low and reproducibility is poor. Butyl chloroformate and dimethyl sulfite can also be used for derivatisation in water, and acetic anhydride can be used for in-situ derivatisation of phenolic herbicides. For increasing selectivity of GC detection pentafluorobenzyl bromide (for ECD) and (2 cyanethyl)dimethyl(diethylamino)silane (for NPD) have been applied. Very characteristic ions are produced in mass spectra if silyl groups are introduced, e.g. by using bis(trimethylsilyl)trifluoroacetamide. Tetramethylammonium, trimethylphenylammonium, tetraalkylammonium, and trimethylsulfonium hydroxides and salts can by used for derivatisation at elevated temperature in the GC injection port. Extractive alkylation is relatively efficient if tetraalkylammonium salts with long chain alkyl groups are used. PMID- 12904960 TI - Separation of the enantiomers of substituted dihydrofurocoumarins by HPLC using macrocyclic glycopeptide chiral stationary phases. AB - Enantiomer separations by HPLC using the macrocyclic glycopeptides teicoplanin (Chirobiotic T), teicoplanin aglycon (Chirobiotic TAG), and ristocetin A (Chirobiotic R) chiral stationary phases (CSP) have been achieved on a unique series of potentially biologically active racemic analogues of dihydrofurocoumarin. The macrocyclic glycopeptides have proven to be very selective for this class of compound. All of the 28 chiral analogues examined afforded baseline separation on at least one of the macrocyclic glycopeptide CSP. The teicoplanin CSP showed the broadest enantioselectivity with 24 of the compounds baseline separated. The TAG and the R CSP produced 23 and 14 baseline separations respectively. All three mobile phase modes, i.e. normal phase (NP), reversed phase (RP), and new polar organic modes (PO), have been evaluated. The NP mode proved to be most effective for the separation of chiral dihydrofurocoumarins on all CSP tested. In the reversed phase (RP) mode, all three CSP separated a similar number of compounds. It was observed that the structural characteristics of the analytes and steric effects are very important factors leading to chiral recognition. Hydrogen bonding was found to play a secondary role in chiral discrimination in the normal phase and polar organic modes. Hydrophobic interactions are important for chiral separation in the reversed-phase mode. Chromatographic retention data does not provide information on the absolute configuration of these chiral dihydrofurocoumarin derivatives. However, when coupled with circular dichroism using the exciton coupling chirality method, the enantiomer elution order and the absolute configuration of some chiral dihydrofurocoumarins were successfully determined. PMID- 12904962 TI - TIMS versus multicollector-ICP-MS: coexistence or struggle for survival? PMID- 12904961 TI - Trace element speciation by ICP-MS in large biomolecules and its potential for proteomics. AB - Latest studies on the chemical association of trace elements to large biomolecules and their importance on the bioinorganic and clinical fields are examined. The complexity of the speciation of metal-biomolecules associations in various biological fluids is stressed. Analytical strategies to tackle speciation analysis and the-state-of-the-art of the instrumentation employed for this purpose are critically reviewed. Hyphenated techniques based on coupling chromatographic separation techniques with ICP-MS detection are now established as the most realistic and potent analytical tools available for real-life speciation analysis. Therefore, the status and potential of metal and semimetals elemental speciation in large biocompounds using ICP-MS detection is mainly focused here by reviewing reported metallo-complexes separations using size exclusion (SEC), ion-exchange (IE), reverse phase chromatography (RP) and capillary electrophoresis (CE). Species of interest include coordination complexes of metals with larger proteins (e.g. in serum, breat milk, etc.) and metallothioneins (e.g. in cytosols from animals and plants) as well as selenoproteins (e.g. in nutritional supplements), DNA-cisplatin adducts and metal/semimetal binding to carbohydrates. An effort is made to assess the potential of present trace elements speciation knowledge and techniques for "heteroatom-tagged" (via ICP-MS) proteomics. PMID- 12904963 TI - Assessment of element-specific homogeneity in reference materials using microanalytical techniques. AB - The IAEA established in 1994 a co-ordinated research programme (CRP) on "Reference Materials for Microanalytical Nuclear Techniques" as part of its efforts to promote and strengthen the use of nuclear analytical technologies in member states with the specific aim of improving the quality of analysis of nuclear, environmental, and biological materials. The objectives of this initiative were: to identify suitable biological reference materials which could serve the needs for quality control in microanalytical techniques; to evaluate existing CRMs for use in microanalytical investigations; to evaluate appropriate sample pretreatment procedures for materials being used for analysis with microanalytical techniques; to identify analytical techniques which can be used for characterisation of homogeneity determination, and to apply such techniques to the characterization of candidate reference materials for use with microanalytical techniques. The CRP lasted for 4 years and seven laboratories and the Agency's Laboratories in Seibersdorf participated. A number of materials including the candidate reference materials IAEA 338 (lichen) and IAEA 413 (single cell algae, elevated level) were evaluated for the distribution of elements such as Cl, K, Ca, Cr, Mn, Fe, Zn, As, Br, Rb, Cd, Hg, and Pb. The results obtained during this CRP suggest that: each element exhibits its characteristic distribution in a matrix described by the "Ingamels' sampling constant" or the "relative homogeneity factor" of Kurfuerst; both concepts are valid over a large range of sample mass used for analysis (from 0.1 microg to around 100 mg); and materials being characterised quantitatively for element homogeneity could be used for the experimental determination of total uncertainty of other analytical techniques. As far as we are aware this is the first time the concept of quantitative characterisation of homogeneity has been applied to potential reference materials and the first demonstration of the feasibility and usefulness of the concept with particular emphasis on enhancing quality control opportunities for microanalytical techniques. PMID- 12904965 TI - Agomelatine(S 20098) antagonizes the penile erections induced by the stimulation of 5-HT2C receptors in Wistar rats. AB - Agomelatine, an antidepressant with melatonin agonist and 5-HT(2C) antagonist properties, as well as two of its main metabolites, S 21517 (N-[2-(7-hydroxy-1 naphtyl)ethyl]acetamide) and S21540 (N-[2-(3-hydroxy-7-methoxynaphtalen-1 yl)ethyl]acetamide), have been assessed in vitro on pig choroid plexus preparations to determine their affinities for 5-HT(2C) receptors and their effects on inositol phosphate production. These compounds were also tested for their ability to inhibit the penile erections induced by the 5-HT(2C) receptor agonists, m-(chlorophenyl)piperazine (mCPP, 0.75 mg/kg, SC) and Ro 60-0175 (2.5 mg/kg, SC) in Wistar rats. These in vivo effects were compared to those of melatonin and the 5-HT antagonists pizotifen and SB 206,553. Agomelatine and S 21517 had moderate affinity for 5-HT(2C) receptors and behaved in vitro as weak antagonists at this receptor subtype. S 21540 had a 10-fold lower affinity. Pizotifen and SB 206,553 antagonized mCPP- and Ro 60-0175-induced penile erections, suggesting that penile erections induced by mCPP or Ro 60-0175 resulted from the stimulation of 5-HT(2C) receptors. Whereas increasing doses (from 1.25 to 40 mg/kg, IP) of melatonin were unable to modify the penile erections induced by mCPP and Ro 60-0175, agomelatine (from 1.25 to 40 mg/kg, IP) dose-dependently decreased mCPP- as well Ro 60-0175-induced penile erections. Furthermore, increasing doses (from 1.25 to 40 mg/kg, IP) of S 21517 and S 21540, the two main metabolites of agomelatine, did not affect the penile erections induced by mCPP and Ro 60-0175. Considering the similar activity of melatonin and agomelatine at melatonin receptors, these data suggested that the reported effects were not due to the stimulation of melatonin receptors and that, contrary to melatonin, agomelatine exerted 5-HT(2C) receptor antagonist properties in addition to its agonist activity at melatonin receptors. Finally, neither S 21517 nor S 21540 seemed to participate to the observed inhibition of penile erections by agomelatine. PMID- 12904966 TI - Effect of antipsychotics on cortical inhibition using transcranial magnetic stimulation. AB - Previous studies suggest that antipsychotic medications may alter cortical inhibition (CI). The current study was designed to determine if typical or atypical antipsychotics indeed alter CI in healthy subjects using three CI paradigms as measured with transcranial magnetic stimulation (TMS): short interval intracortical inhibition (SICI), cortical silent period (CSP) and transcallosal inhibition (TCI). CI was measured before, 6 and 24 h after being randomly assigned to receive a single dose of 2 mg haloperidol (n=8), 10 mg olanzapine (n=10) or placebo (n=9). There was no significant effect on any measure of CI at 6 and 24 h after receiving olanzapine, haloperidol or placebo. Moreover, no significant change in the motor threshold was observed across the three medication groups. Therefore, single administration of an antipsychotic has no effect on CI or resting motor threshold. Whether chronic, repeated administration of antipsychotics has effects on CI requires further investigation. PMID- 12904967 TI - The influence of changing nicotine to tar ratios on human puffing behaviour and perceived sensory response. AB - RATIONALE: Smokers modify their smoking behaviour when switching from their usual product to higher or lower tar and nicotine-yield cigarettes. OBJECTIVE: The aims of the current study were to assess the influence of varying nicotine yields at constant tar yield on human puffing measures, nicotine deliveries under human smoking conditions and the sensory response to mainstream cigarette smoke. These assessments would allow an evaluation of the degree of compensation and the various possible causes of changes, if any. METHODS: The participants were 13 regular smokers of commercial or hand-rolled cigarettes. They were tested with four cigarettes, which exhibited a wide range of nicotine to "tar" ratios at a relatively constant "tar" yield. Their smoking behaviour was monitored by placing the test cigarettes into an orifice-type holder/flowmeter attached to a custom built smoker behaviour analyser. In addition, a comprehensive sensory evaluation of the products was carried out. RESULTS: The differences in the nicotine to tar ratios of the samples did not significantly influence the puffing behaviour patterns, i.e. puff number and interval, total and average puff volume, integrated pressure and puff duration. Additionally the pre- to post-exhaled CO boosts were not significantly influenced by the experimental samples used in the study. However, the nicotine yields obtained by the smokers were significantly influenced by the machine-smoked nicotine yields or the nicotine to tar ratios of the samples. The machine-smoked nicotine yields were highly correlated with the nicotine yields obtained under human smoking conditions. For the sensory evaluation, there was only a significant difference between the samples in the intensity of the impact. CONCLUSION: These observations imply that these puffing variables are not controlled by the nicotine yield of the cigarette. PMID- 12904968 TI - The 5-HT2A receptor antagonist M100,907 attenuates motor and 'impulsive-type' behaviours produced by NMDA receptor antagonism. AB - In the present series of studies, we have investigated the effects of antagonists selective for the 5-HT(2A), 5-HT(2B) and 5-HT(2C) receptors on motor and 'impulsive'-type behaviours elicited by the non-competitive N-methyl- d-aspartate (NMDA) antagonist dizocilpine. The selective 5-HT(2A) receptor antagonist M100,907 (0.5 mg/kg) attenuated the hyperlocomotion and stereotypy produced by dizocilpine (0.1-0.3 mg/kg). The selective 5-HT(2B) receptor antagonist SB215,505 (3 mg/kg) and the selective 5-HT(2C) receptor antagonist SB242,084 (0.5 mg/kg) had no effect against either measure, except that SB242,084 produced a small potentiation of the hyperactivity response. Dizocilpine (0.03 mg/kg) increased premature responding in rats performing the 5-choice serial reaction time task (5 CSRTT), and increased response frequency consequently reducing the mean inter response time (IRT) and efficiency of responding in a DRL24 task. M100,907 (0.5 mg/kg) attenuated each of these effects, as well as the increased premature responding produced by the NMDA NR2B selective antagonist Ro 63-1908 (1 mg/kg) in the 5-CSRTT. In contrast SB242,084 (0.5 mg/kg) did not attenuate the dizocilpine induced premature responding or increased responding in the DRL24 task. Rather, SB242,084 (0.05-0.5 mg/kg) produced qualitatively similar effects to dizocilpine, increasing premature responding and reducing IRT. The results suggest that 5 HT(2A) receptor antagonists may normalise certain 'impulsive' behaviours produced by NMDA receptor hypofunction. The 5-HT(2C) receptor antagonist SB242,084 failed to exert equivalent effects, rather a trend toward exacerbation of the behavioural changes produced by dizocilpine was apparent. PMID- 12904969 TI - Expression of the VEGF receptor-3 in osteoarthritic chondrocytes: stimulation by interleukin-1 beta and association with beta 1-integrins. AB - Recent studies have demonstrated enhanced expression of vascular endothelial growth factor and vascular endothelial growth factor receptor (VEGFR)-1 and -2 in chondrocytes of rheumatoid and osteoarthritic cartilage. Since expression of VEGFR-3 ( Flt-4) in chondrocytes has not yet been investigated, we studied the distribution of VEGFR-3 in osteoarthritic cartilage samples by immunohistochemistry and immunoelectron microscopy. Furthermore, we looked for functional colocalization of VEGFR-3 with the signal transduction receptor beta(1)-integrin. Superficial osteoarthritic chondrocytes exhibited VEGFR-3 expression in the cytoplasm and on the cell membrane. Using western blotting we could demonstrate that interleukin-1 beta (IL-1 beta) stimulates the expression of VEGFR-3 in chondrocytes in vitro in a dose-dependent manner. By coimmunoprecipitation assay we found a functional complex between the beta(1) integrin and VEGFR-3 in IL-1 beta-stimulated chondrocytes indicating that activated VEGFR-3 may interact with beta(1)-integrin and associated subcellular pathways in osteoarthritic chondrocytes. Taken together with results of previous studies showing that beta(1)-integrins were also associated with other surface receptors and proteins in chondrocytes that cause cartilage destruction in arthritis (for example, urokinase-type plasminogen activator receptor and matrix metalloproteinases), we can hypothesize that signal transduction by these receptor complexes via beta(1)-integrins may play a crucial role in pathogenesis of osteoarticular disorders. Further work needs to be done to elucidate downstream signaling events activated by these receptors. PMID- 12904970 TI - Close apposition of dynorphin-positive nerve fibres to lymphocytes in the liver suggests opioidergic neuroimmunomodulation. AB - The liver is innervated by sympathetic efferent, spinal afferent, vagal afferent and probably also vagal efferent fibres. To assess potential functional roles of the various neuronal subsets, data on transmitter systems are of crucial importance. This study was aimed at elucidating a possible opioidergic system in the mouse and rat liver. In particular relationships of opioidergic neurons to immune cells were emphasised. Material from perfusion-fixed mice ( n=29) of different strains (BALB/c, NMRI, C57Bl6, SV 129 inbred) and Wistar rats ( n=7) was cryosectioned at 12-14 microm and incubated for single or double immunofluorescence. Antibodies directed against dynorphin A, met-enkephalin, endomorphin 1 and 2, mu, kappa- and delta-opioid receptors (MOR, KOR, DOR), tyrosine hydroxylase (TH), dopamine beta-hydroxylase (DBH), CD4, CD8 and macrophages were used. Binding sites were detected using Cy3-, FITC-, DTAF-, Cy2 , Alexa 555- and Texas red-tagged secondary antibodies. Specimens were analysed using confocal laser scanning microscopy (CLSM). Numerous nerve fibres staining for dynorphin were found in periportal areas of both mouse and rat livers. Neither met-enkephalin nor endomorphin could be detected in axons. No immunopositive neuronal cell bodies or other cellular elements were seen. All dynorphin positive fibres costained for TH while not every TH-positive fibre costained for dynorphin. Thus, most if not all dynorphin-positive nerve fibres may be of sympathetic origin. KOR immunostaining could be localised to round mononuclear cells which often costained for CD4, less frequently for CD8 and rarely for the pan-macrophage marker BM8. Altogether, about 45% of KOR-positive cells were identified as T-lymphocytes. In some instances, close appositions of dynorphin-positive axons to KOR-positive cells were revealed by CLSM. No KOR immunoreactivity was detected in nerve fibres. Hence, sympathetic neurons innervating the liver may interfere with inflammatory processes, in addition to their well-established beta(2)-adrenergic effect, via an opioidergic action on immune cells. PMID- 12904971 TI - Estrogen receptor expression in meibomian glands and its correlation with age and dry-eye parameters. AB - PURPOSE: To investigate the correlation between estrogen receptor-positive basal cells of the meibomian glands of the lower lid with age, gender, and subjective and objective dry-eye parameters. METHODS: Sixteen lower lid specimens were collected from 7 female and 9 male patients (age range 22-88 years) during tumor surgery requiring whole-thickness excision. Prior to surgery the patients were asked about any dry-eye symptoms (score 0-4, where 0=no dry-eye symptoms). Tear break-up time was measured, and Schirmer I and II tests were performed. We obtained histological sections from the outer margins of the formalin-fixed and paraffin-embedded specimens and performed immunohistochemical staining for estrogen, progesterone and androgen receptors. At least 500 cells were counted per specimen, and the proportion of positively stained cells was calculated and correlated to the age and dry-eye parameters. RESULTS: All meibomian glands had positive nuclear staining with antibodies for estrogen receptors in their outer cell layers, i.e., the basal cell layer. The proportion of cells expressing estrogen receptors increased with age independent of gender (correlation coefficient=0.67, P<0.005). No correlation was found between estrogen receptor positivity and subjective dry-eye symptoms, tear break-up time, or Schirmer I and II test results. There was no difference in the proportion of cells expressing estrogen receptors between female and male patients (P=0.5). CONCLUSIONS: The amount of meibomian gland cells expressing estrogen receptors in the lower lid seems to increase with age independent of gender and seems not to affect the fat layer and stability of the tear film. PMID- 12904972 TI - Y-chromosomal STR haplotypes in a Romanian population sample. AB - A set of seven STR-loci mapping on the male-specific region of the human Y chromosome (DYS19, DYS385, DYS389 I/II, DYS390, DYS391, DYS392, DYS393) were typed by means of two multiplex PCR reactions and capillary electrophoresis in a Romanian population sample of 104 unrelated males. Among the 97 different haplotypes observed, 92 were unique, while 5 occurred more than once. The observed haplotype diversity was 0.989. PMID- 12904973 TI - Spanish population data and forensic usefulness of a novel Y-STR set (DYS437, DYS438, DYS439, DYS460, DYS461, GATA A10, GATA C4, GATA H4). AB - DNA typing of 8 recently described STRs on the Y chromosome was carried out by means of 2 multiplex amplification reactions for 134 unrelated males from Cantabria, a region in northern Spain. Multiplex 1 included loci DYS460 (GATA A7.1), GATA A10, GATA H4 and DYS439; multiplex 2 included DYS461 (GATA A7.2), GATA C4, DYS437 and DYS438. Haplotype diversity was found to be 99.36%, similar to that obtained with the standard 9-STR set ("minimal haplotype") of the European Y-user group (99.35%). The 13-locus haplotype resulting from the combination of the standard minimal haplotype and the 4-locus multiplex 1 showed a 99.89% diversity. Further inclusion of the 4 loci in multiplex 2 resulted in a haplotype diversity of 99.93%. The combination of the "minimal haplotype" and the multiplex 1 in the present study may be an efficient way of increasing the power of discrimination in forensic cases. PMID- 12904974 TI - Lithium in the acute treatment of bipolar disorders-a stocktaking. AB - Before the rise of atypical antipsychotics, lithium used to be the most frequently investigated substance in the acute treatment of bipolar disorders, although studies are not always of the highest methodological standard. Due to the doubt about a sufficient efficacy of lithium expressed in recent years from various sides, and the simultaneous availability of newer treatment alternatives, this paper attempts to make a critical stocktaking of our knowledge about lithium in the acute treatment of bipolar disorders. Aspects concerning the changed disorder concept through the broadening of the bipolar spectrum, together with the available results from controlled and open studies with lithium, are presented and appraised. This shows that lithium should still be seen as an essential, but not the only corner stone in the differentiated treatment of bipolar patients. Provided that it is taken reliably and well-tolerated, lithium represents a first choice treatment, particularly for a classical course of manic depressive illness (Bipolar I disorder), especially for mild to moderate manic syndromes.However, as antidepressive treatment, lithium should rather not be applied as a monotherapy, particularly in severe bipolar depression, since with the new generation of antidepressants and anticonvulsants well-tolerated and very effective alternatives are available. In combination treatment, lithium should be applied particularly when it has already shown good prophylactic efficacy and/or in patients for whom a high suicide risk must be presumed. PMID- 12904975 TI - Lithium in the long-term treatment of bipolar disorders. AB - Usefulness of lithium in the prophylaxis of bipolar disorders has been challenged for five major reasons. The authors review the empirical basis of these criticisms and come to the following conclusions. 1. Lithium efficacy is high and beyond reasonable doubt in classic manic-depressive illness. Bipolar patients presenting atypical features show a much poorer response rate to lithium. 2. There is no empirical evidence for a loss of lithium efficacy over time. 3. There is little evidence for discontinuation-induced refractoriness to lithium. 4. Lithium withdrawal phenomena are well established but seem to be rather specific to certain subgroups. Withdrawal phenomena seem to be common in atypical bipolar disorder but rare in fully stabilized classic manic-depressive illness. 5. Other factors limiting lithium efficacy in clinical practice (e. g., non-compliance) are not specific to lithium. In conclusion, prophylactic lithium does have major drawbacks and there is a clear need for more efficacious alternatives in non classic bipolars. Compared to existing alternatives, lithium currently is to be considered the golden standard. This status might, however, be challenged by major alternative mood-stabilizers that are presently under clinical investigation. PMID- 12904976 TI - How should findings on antisuicidal effects of lithium be integrated into practical treatment decisions? AB - Beyond its prophylactic efficacy lithium has demonstrated possibly specific antisuicidal effects. Lithium significantly reduces the high excess mortality of patients with affective disorders. Appropriate lithium prophylaxis prevents ca. 250 suicides per year in Germany although lithium salts are prescribed within the National Health Scheme at low frequency (0.06% of the population). Rational treatment strategies most likely would demand for about 10 times higher prescription rates. Guidelines and algorithms for selecting an appropriate prophylactic strategy in affective disorders should take into consideration the suicide risk of an individual patient. PMID- 12904977 TI - Lithium augmentation therapy in refractory depression-update 2002. AB - Lithium has been used to augment the efficacy of antidepressant medications for more than 20 years. The present study examines whether evidence exists to support the clinical efficacy of lithium augmentation in refractory, treatment resistant depression. Studies were identified by searching Medline (1980 to August 2002) and by scanning the references of published reviews and standard textbooks. Studies were selected if they were open-labeled or double-blind, placebo controlled or comparator trials that involved patients who had not responded to conventional antidepressants. 27 prospective studies were identified that included a total of 803 depressed patients displaying the following designs: 10 double-blind, placebo-controlled trials, 2 randomized, double-blind comparator trials, 2 randomized, open comparator trials, and 13 open-label trials. The majority of randomized controlled trials has demonstrated substantial efficacy of lithium augmentation in partial and non responders to antidepressant treatment. In the placebo-controlled trials, the response rate in the lithium group was 45% and in the placebo group 18% (p<0.001). Summarizing all open and controlled studies, approximately 50% of patients responded to lithium augmentation within 4 weeks. In conclusion, lithium is the foremost and most well-documented augmentation strategy in refractory depression.Therefore, it should be considered a first-line treatment strategy in patients with major depression who do not adequately respond to standard antidepressants. PMID- 12904978 TI - St. John's Wort. An option for the primary care treatment of depressive patients? AB - This article reviews the historical background, active ingredients of St. John's Wort and the major double-blind placebo-controlled studies. Despite the two major failed clinical trials conducted in American Research Centers, most of the data reviewed support that hypericum extracts are more effective than placebo for the treatment of mild to moderate depressive illness. The authors examine the likely reasons for the failed studies and also describe drug interactions and the side effects of St. John's Wort. Those patients who are prescribed St. John's Wort should be closely monitored. PMID- 12904979 TI - Is recurrent brief depression an expression of mood spectrum disorders in young people? Results of a large community sample. AB - The clinical relevance of Recurrent Brief Depression (RBD) has not received sufficient attention to date and continues to represent a controversial issue. The present study was carried out in a community sample to evaluate the lifetime prevalence of RDB, the degree of comorbidity, as well as possible risk factors. Subjects from a community survey in Sardinia (Italy) were randomly selected from registers of a rural, an urban and a mining area (n=1040, 461 males, 579 females). Interviews were carried out by physicians using the Italian version of the Composite International Diagnostic Interview Simplified which had been modified for the purpose of this study. Lifetime prevalence of RBD was 7.6%; 5.8% in males, 9% in females. Subjects aged 18 to 24 years presented higher frequencies (13.8%, OR 2.2) than those aged 25 or over. Comorbidity with Major Depression was particularly frequent. RBD was furthermore associated with suicide attempts and substance abuse, thereby constituting an effective health problem. Further epidemiological and clinical studies of RBD are warranted in order to develop specific treatments and prevention strategies. PMID- 12904980 TI - Cerebral networks linked to the event-related potential P300. AB - P300 is an event-related potential that is elicited by an oddball paradigm. In several neuropsychiatric diseases, differences in latencies and amplitude compared to healthy subjects have been reported. Because of its clinical significance, several investigations have tried to elucidate the intracranial origins of the P300 component. In the present study we could demonstrate a network of P300 generators. Investigated were 15 healthy subjects with an acoustical oddball paradigm within a fMRI block design, which enabled us to exclude attention or acoustical processing effects. The inferior and middle frontal, superior temporal, lower parietal cortex, the insula and the anterior cingulum were significantly activated symmetrical in both hemispheres. PMID- 12904981 TI - Wrist fracture in a patient undergoing electroconvulsive treatment monitored using the "cuff" method. AB - We report on a case of a 64-year-old female patient who received electroconvulsive treatment for major depression. The patient sustained a fracture to the distal radius of the arm blocked by the "cuff" method (Smith fracture). Subsequent investigations revealed that she was suffering from osteoporosis. This case report underlines the importance of special treatment procedures for osteoporotic patients and other patient groups at risk for fractures. We recommend screening for osteoporosis in routine pre-ECT evaluation, even in younger patients, and propose not attaching the cuff to the arm in those at risk for fractures. In our patient, electroconvulsive therapy was safely continued using higher doses of succinylcholine and without the cuff method. PMID- 12904983 TI - Partial cricotracheal resection for pediatric subglottic stenosis: a single institution's experience in 60 cases. AB - In our study, 60 infants and children, each with a severe subglottic stenosis (SGS), underwent partial cricotracheal resection (PCTR) with primary thyrotracheal anastomosis. According to the Myer-Cotton classification, two were grade II, 41 were grade III and 17 were grade IV stenoses. Of the 60 patients, 57 (95%) are presently decannulated, and one patient sustained a complete restenosis. Two patients with better than 80% subglottic airways still are waiting for decannulation: one because of bilateral cricoarytenoid joint fixation and the second because of temporary stenting of the subglottis with a Montgomery T-tube. The rate of decannulation is 97% (36 of 37 cases) in primary PCTRs, 100% (13 of 13 cases) in salvage PCTRs for failed laryngotracheal reconstructions (LTR) and 70% (7 of 10 cases) in extended PCTRs (i.e., PCTR associated with an additional open-airway procedure). PMID- 12904985 TI - Successful treatment of primary non-Hodgkin's lymphoma of the vagina with chemotherapy. AB - INTRODUCTION: The vagina as a primary site of malignant lymphoma in woman is extremely rare. CASE REPORT: We report here a 30-year-old patient with a primary vaginal non-Hodgkin's lymphoma stage IEA, follicular large cell of B lineage, with an excellent response to cytotoxic chemotherapy (CHOP) and event-free survival of 28 months. CONCLUSION: Primary involvement of the vagina can be successfully treated by pelvic irradiation, but in young women chemotherapy should be considered to preserve fertility, as well as quality of life. PMID- 12904984 TI - Double tumor of the larynx: a case report. AB - A case is described of a double laryngeal tumor with simultaneous evolution but different histological patterns: a chondrosarcoma and an epidermoid carcinoma "in situ" involving both vocal cords. To our knowledge, only one previous case has been reported in the literature. PMID- 12904987 TI - Pregnancy complications, obstetric risks, and neonatal outcome in singleton and twin pregnancies after GIFT and IVF. AB - PURPOSE: In vitro fertilization (IVF) and to a lower extent gamete intra fallopian transfer (GIFT) have become routine infertility treatments in industrialized countries. Our purpose is to compare the obstetric and neonatal characteristics of singleton and twin pregnancies after GIFT and IVF with those conceived spontaneously. METHODS: This case-control study was conducted in a tertiary care medical center. The 322 singleton and 78 twin pregnancies after GIFT or IVF from 1991 through 1996 were evaluated and compared with each other, and with a control group that conceived spontaneously and matched for parity, maternal and gestational age. Statistical significance of differences was assessed by chi(2) test or two-tailed Fisher exact test. Continuous variables were compared by the paired t-test. RESULTS: Pregnancy-induced hypertension (PIH) and vaginal bleeding were significantly more frequent maternal complications in the GIFT/IVF singleton groups compared to controls. In twin pregnancies the rate of cesarean sections, vaginal bleeding and preterm labor were more common after GIFT/IVF but did not reach statistical significance. Assisted reproduction was associated with low birth weight only in twin pregnancies when controlled for confounding variables, however perinatal outcome was comparable. There was no significant difference in the outcome measures between GIFT and IVF pregnancies. CONCLUSION: After controlling for parity, maternal and gestational age, singleton pregnancies conceived by GIFT/IVF are at increased obstetrical risk, however the perinatal outcome is comparable despite a lower average birth weight. PMID- 12904989 TI - Shortening of the patellar tendon length does not influence the patellofemoral alignment in a cadaveric model. AB - INTRODUCTION: Anterior cruciate ligament (ACL) reconstruction with the use of autograft tissue represents the standard treatment. The use of a bone-patellar tendon-bone transplant for symptomatic ACL deficiency achieves good long-term results. The purpose of the study was to investigate in a cadaveric model whether reproducible patellar tendon shortening changes the patellofemoral alignment. MATERIALS AND METHODS: Using five cadaveric knees, an MRI investigation was performed with the patellar tendon left unchanged, shortened 5 mm and shortened 10 mm, respectively, in both 20 and 45 degrees of knee flexion. The lateral patellofemoral and the congruence angles were measured and compared using a one way analysis of variance for repeated measurements. RESULTS: Shortening the patella tendon by approximately 20% did not significantly influence the patellofemoral alignment. CONCLUSION: Although anterior cruciate ligament reconstruction using the patellar tendon has become a standard procedure, postoperative problems such as anterior knee pain, and patellofemoral osteoarthritis occur quite often. In this cadaver study we did not find a significant difference for the patellofemoral alignment, irrespective of the patellar tendon length. PMID- 12904990 TI - Expression of serine proteases and metalloproteinases in human pituitary adenomas and anterior pituitary lobe tissue. AB - Data on the dural invasiveness of pituitary adenomas have been correlated to the expression of matrix metalloproteinases (e.g. MMP-9). Serine proteases have not yet been investigated in human pituitary adenomas. In this study, paraffin embedded material from 84 human pituitary adenomas (acromegaly n=18, Cushing's disease n=21, prolactinoma n=18, thyroid-stimulating hormone-secreting adenoma n=1, nonsecreting adenoma n=26) and 9 nontumourous anterior pituitary lobes (obtained from patients with prostate cancer) was immunohistochemically analysed for expression of MMP-2, MMP-9, tissue inhibitor of metalloproteinases-2 (TIMP 2), urokinase-type plasminogen activator (uPA), uPA receptor (uPAR), tissue-type plasminogen activator (tPA), plasminogen activator inhibitor-1 (PAI-1), and interleukin-6 (IL-6). Cavernous sinus invasion was determined by assessment of preoperative magnetic resonance imaging and intraoperative inspection (invasive n=50, noninvasive n=34). In pituitary adenomas, reactions were positive (diffuse expression) to MMP-2 (74% of cases), MMP-9 (49%), TIMP-2 (88%), uPA (89%), uPAR (90%), tPA (69%), and PAI-1 (87%). A weak expression of IL-6 was found in 12% of the adenomas. All reactions were positive (focal expression) in every sample of anterior lobe tissue, except for uPA (negative in 3 out of 9 cases), and IL-6 (faintly positive in 5 out of 8 cases). Adenomas showed remarkably greater expression of uPA than anterior lobe tissue (Chi-square P<0.05). Nonsecreting adenomas exhibited a stronger tendency towards overexpression of uPA in invasive tumours when compared to noninvasive adenomas (Chi-square P=0.053). We found no correlation of MMP-9 expression and tumour invasion. TIMP-2 was overexpressed in noninvasive as compared to invasive adenomas (Chi-square P<0.05). The interrelationship between MMPs and serine proteinases in pituitary adenomas remains to be elucidated. From our data, a correlation between IL-6 and an activation of MMP-9 cannot be proven. The uPA-system may, however, play a role in dural invasion of pituitary adenomas. PMID- 12904991 TI - PTEN methylation and expression in glioblastomas. AB - The tumor suppressor gene PTEN on chromosome 10q23.3 regulates the Akt signaling pathway and modulates cell growth and apoptosis. The PTEN gene is mutated in 20 40% of glioblastomas. In this study, we assessed whether loss of PTEN expression is also caused epigenetically. Methylation-specific PCR revealed that CpG islands of the PTEN promoter were methylated in 27 of 77 (35%) glioblastomas and in 4 of 11 (36%) glioblastoma cell lines. Only two glioblastomas showed loss of PTEN immunoreactivity in the entire biopsy; both had a missense PTEN mutation and LOH at the PTEN locus, but lacked PTEN methylation. In biopsy specimens with focal loss of PTEN expression, DNA samples extracted from microdissected foci showed PTEN methylation only in areas with loss of PTEN expression. These results suggest that PTEN methylation occurs frequently in glioblastomas and may be associated with focal loss of PTEN expression. However, the correlation between PTEN methylation, PTEN mutations, LOH at the PTEN locus, and loss of PTEN protein expression was inconsistent. Possible reasons for discrepancies between gene status and protein expression include differences in the biological effect of specific PTEN mutations and the possibility that the processed PTEN pseudogene on 9p21 is expressed in glioblastomas and co-reacts with the PTEN antibody. PMID- 12904992 TI - Age-associated prevalence and risk factors of Lewy body pathology in a general population: the Hisayama study. AB - In dementia with Lewy bodies (DLB), the Lewy bodies (LBs) are an essential substrate. Although LB pathology has gained increasing attention as one of the major causes of dementia, little is known about the exact prevalence of LB pathology in the general population. In addition, the pathology of Alzheimer-type dementia (ATD) is frequently associated with DLB. To investigate the prevalence of LB pathology in a community-based population and to evaluate the relationship between LB and ATD pathology, we performed an analysis of 102 consecutive autopsy cases. The survey extended over 2.5 years and autopsy rate was 70.5%. LB pathology was detected using alpha-synuclein immunohistochemistry and was assessed based on consensus guidelines for DLB. ATD pathology was evaluated by both CERAD and NIA-RI criteria. Twenty-nine subjects were clinically demented. LB pathology was present in 23 (22.5%) of 102 cases, and in 12 (41.4%) of the demented subjects. The LB score was not significantly different between DLB cases and non-demented subjects with LB pathology (nd-LB), while the Braak stages were significantly different between the two groups. Prevalence of LB pathology constantly increased with age. DLB cases accompanying severe ATD pathology showed more rapid increase of LB scores than did DLB cases without severe ATD pathology. Moreover, DLB cases with severe ATD pathology had poorer prognoses than those without severe ATD pathology. Our results suggested that aging and severe ATD pathology have a strong effect on the evolution of LB pathology. PMID- 12904993 TI - Age-associated prevalence and risk factors of Lewy body pathology in a general population. PMID- 12904994 TI - Microvascular damage following experimental sinus-vein thrombosis in rats. AB - We evaluated effects of thrombosis of the superior sagittal sinus, its bridging and cortical veins (SVT) on the cerebral microvasculature in rats. Cryosections of brains ( n=7) were examined for venous infarction and microvascular basal lamina damage 3 h after SVT by immunohistochemical staining of microtubule associated protein 2 and collagen type IV. Microvessels in the infarctions showed a decrease in the number (23.5+/-6.1%, P<0.002) and the total area (24.9+/-6.5%, P<0.011) of collagen type IV-positive vessels in contrast to control areas (21.7+/-12.4%, P<0.007; and 26.3+/-15.1%, P<0.026 in contrast to control areas of unoperated animals). This study showed a significant alteration of the cerebral microvasculature in SVT, which might contribute to edema and hemorrhagic transformation. PMID- 12904995 TI - Rectal cancer: how accurate can imaging predict the T stage and the circumferential resection margin? AB - BACKGROUND: One of the concerns after rectal cancer surgery is the local recurrence rate. Randomized trials have shown that the best local control rate for rectal cancer patients overall is obtained by a short preoperative course of radiotherapy followed by a total mesorectal excision, but there are however subgroups of rectal cancer patients with varying risks of recurrences. DISCUSSION: At one end of the continuum are patients with stage I disease who are at very low risk of local recurrence. At the other end are the patients with locally advanced tumor who are at very high risk, and who would benefit from a more extensive neoadjuvant treatment schedule. This kind of differentiated treatment can be performed only when there is an accurate tool to identify preoperatively the different risk groups. There is recent evidence, suggesting that MRI can serve for this purpose, because it accurately predicts the tumor free circumferential resection margin at TME. CONCLUSION: This article provides a literature review of the value of preoperative imaging in rectal cancer surgery. PMID- 12904997 TI - Polymorphism of monocyte chemoattractant protein 1 in Crohn's disease. AB - BACKGROUND AND AIMS: The chemokine MCP-1 is thought to be important for the recruitment of mononuclear cells and the maintenance of inflammation in inflammatory bowel disease. We investigated whether MCP-1 protein expression is correlated with the degree of mucosal inflammation in patients with Crohn's disease. Furthermore, we studied whether a functional single nucleotide polymorphism (G or A) located in the distal regulatory region of the MCP-1 gene is associated with Crohn's disease and/or its phenotype. PATIENTS AND METHODS: MCP-1 concentration in tissue homogenates was analyzed in mucosal biopsy specimens of 31 patients with Crohn's disease and 48 controls by enzyme-linked immunosorbent assay, and the correlation with an endoscopic macroscopic score was analyzed. In 179 patients with Crohn's disease and 189 controls MCP-1 genotyping was carried out by polymerase chain reaction restriction fragment length polymorphism technique. Subgroup phenotypic analysis was performed according to the Vienna classification. RESULTS: MCP-1 tissue concentrations were significantly associated with the macroscopic degree of inflammation. The gene frequency of the different MCP-1 alleles did not differ from healthy controls. However, the G/A and G/G genotype was significantly decreased in patients with a later onset of the disease and both genotypes presented also less frequently with a fistulizing disease behavior. CONCLUSION: The degree of intestinal inflammation in Crohn's disease is associated with MCP-1 tissue levels. Furthermore there is evidence for an association of different disease behavior with different MCP-1 genotypes. PMID- 12904996 TI - Chemoprevention of colorectal cancer in ulcerative colitis. AB - BACKGROUND: Patients with ulcerative colitis (UC) are at greater risk of developing colorectal cancer (CRC) than the general population. Both duration and extent of UC are important risk factors for CRC, as is the presence of primary sclerosing cholangitis, family history of CRC, and (in some studies) early age at diagnosis of UC. Efforts to reduce this risk have focused on colonoscopic surveillance as the best alternative to the more definitive, but less appealing, approach of prophylactic colectomy. However, spurred on by findings in the sporadic CRC literature, there has been a growing interest in a possible role for chemoprevention of CRC in patients with UC. EMPIRICAL STUDIES: Published evidence to date indicates that 5-aminosalicylic acid agents are protective against the development of dysplasia and CRC. Oral, but not topical, steroids also appear to be chemoprotective, but their chronic use cannot be recommended for this indication. Ursodeoxycholic acid has been shown to reduce the risk of neoplasia in UC patients with primary sclerosing cholangitis. Evidence suggests, but does not prove, that folic acid is chemopreventive in patients with UC. Further studies are needed to fully define the chemoprotective role of these and other agents. PMID- 12904999 TI - Combined esophageal and duodenal atresia complicated with meconium peritonitis. PMID- 12904998 TI - No dose-dependent tubulotoxicity of 5-aminosalicylic acid: a prospective study in patients with inflammatory bowel diseases. AB - BACKGROUND AND AIMS: Elevated levels of renal tubular markers in the urine are found in 20-30% of patients with chronic inflammatory bowel diseases. We investigated whether this reflects a dose-dependent tubulotoxicity of 5 aminosalicylic acid (5-ASA). PATIENTS AND METHODS: In an open, prospective, multicenter study 18 patients with Crohn's disease and 29 with ulcerative colitis were treated with 3 g 5-ASA or more daily as the sole drug for 6 weeks. Clinical activity (CDAI, CAI) and renal tubular markers [beta-N-acetyl-D-glucosaminidase (beta-NAG) and other proteins in urine] were monitored. We examined whether the proportion of patients with elevated beta-NAG is more than 15% higher (absolute difference) than that prior to treatment. RESULTS: The proportion decreased from 19.2% to 12.8% in the intention-to-treat analysis (n=47) and from 24.3% to 13.5% in the per-protocol analysis (n=37), which was not more than 15% higher than at baseline. Mean CDAI decreased from 222 to 146 and mean CAI from 7.3 to 3.1 (intention-to-treat analysis). Response to therapy was shown by 61% of patients with Crohn's disease and 66% of patients with ulcerative colitis. The cumulative dose of 5-ASA was not correlated with beta-NAG level in the urine. CONCLUSION: This study largely rules out that 5-ASA at 3 g or higher per day for 6 weeks induces renal tubular damage. Elevated renal tubular markers reflect inflammatory activity or an extraintestinal manifestation of inflammatory bowel diseases. PMID- 12905001 TI - Central venous catheter sepsis in surgical newborns. AB - We examined central venous catheter (CVC) sepsis in newborn surgical patients, as this group appeared to have a higher incidence of this complication. During a 3 year period 79 patients on the surgical service required a tunneled, cuffed, Broviac CVC. Nineteen patients (24% or 9.9 episodes per 1000 catheter days) had proven sepsis and 8 (10% or 1.9/1000 catheter days) had suspected sepsis. An intestinal stoma was definitely related to CVC sepsis ( p<0.001). Other risk factors included lower gestational age, more operations, and younger age at first stoma. Temperature, white blood cell (WBC) count, and platelet count did not correlate with CVC sepsis. We found no better indicator of CVC sepsis than the presentation of an ill child. Certainly temperature, WBC, and platelet count are not reliable indicators. Surgeons have little control over the factors that were found to be related to CVC sepsis. It would appear reasonable from these results to maintain a high index of suspicion in the high-risk groups, to use peripherally inserted central catheters (PIC lines) as the first line of long term vascular access, and to bring CVCs out of a nonabdominal site, perhaps the scalp, in patients with stomas. PMID- 12905002 TI - The successful medical management of severe duodenal strictures secondary to eosinophilic gastroenteritis in an infant. AB - Eosinophilic gastroenteritis is an uncommon condition of unknown etiology that has only been relatively recently reported. Its clinical manifestations range from a mild disease to more severe forms resembling Crohn's disease. Enteric strictures are a rare but recognized complication of this condition. We report a case of eosinophilic gastroenteritis in an infant presenting with severe duodenal strictures that was successfully managed with oral prednisolone. We believe that this is the first reported case of duodenal strictures secondary to eosinophilic gastroenteritis. PMID- 12905003 TI - Sonographic prenatal diagnosis of central nervous system abnormalities. AB - INTRODUCTION: Over the past 20 years, the spectrum of neonatal neurological malformations has changed due to the diffusion of ultrasound, performed either routinely or as required by maternal alpha-fetoprotein screening or history. DISCUSSION: We review and illustrate the potential of ultrasound for the prenatal diagnosis of abnormalities in size or shape of the skull (macrocephaly, microcephaly, craniostenosis), neural tube defects, ventriculomegaly, hydrocephalus, posterior fossa defects (abnormalities in the size of the cisterna magna, cerebellar abnormalities), midline abnormalities (holoprosencephaly, abnormalities of the corpus callosum), ischemic lesions and hemorrhage, tumours, and focalized hyperechogenic images. The limits of fetal ultrasound screening and of the various diagnostic strategies implemented when a fetal brain abnormality is suspected are discussed. Overall, gross lethal abnormalities such as anencephaly or major hydrocephaly are accessible to prenatal sonographic screening, and nearly always result in termination of the pregnancy. However, hydrocephaly may progress late in gestation and remain undiscovered unless a third trimester ultrasound is performed. A majority of cases with myelomeningocele are diagnosed prenatally, resulting either in termination of the pregnancy or in neonatal management. A growing number of more subtle abnormalities, including midline or posterior fossa abnormalities can be spotted by fetal ultrasound, but their postnatal outcome cannot always be predicted accurately, despite the use of fetal magnetic resonance imaging. In such cases, a trans-disciplinary approach involving perinatologists, pediatric radiologists, neuropathologists, neurosurgeons or neurologists familiar with neonates is crucial to counseling the parents. Some brain abnormalities are still extremely difficult or even impossible to diagnose in utero despite advances in sonographic imaging. This is due to the fact that severe neurological impairment may result from conditions that do not affect substantially affect the morphology of the brain, and that major structural abnormalities may develop late in gestation, and thus remain undetected at second trimester ultrasound. CONCLUSION: Ultrasound screening identifies a growing number of central nervous system abnormalities, resulting in substantial changes in the neonatal presentation of neurological congenital abnormalities. PMID- 12905004 TI - Current status of prenatal management of fetal spina bifida in the world: worldwide cooperative survey on the medico-ethical issue. AB - INTRODUCTION: The management of congenital CNS anomalies differs among countries. This paper reports on a worldwide survey of individual experience and regimens for treating spina bifida (SB) in different patient populations and cultural backgrounds. Ten representative members of the International Society for Pediatric Neurosurgery (ISPN) in individual countries registered their experience, prenatal management, and care system for SB. The information collected contained the following major items: intrauterine diagnosis, prophylaxis, and legal termination in prenatal management. DISCUSSION: The prenatal diagnosis of SB is extensively performed throughout the world, but the marker alpha-fetoprotein is not widely accepted in Asia, nor is the prophylactic administration of folic acid, which is not part of the routine care in Asia. Morphological analyses by neuroimaging, especially using an advanced technique of magnetic resonance imaging (MRI), are well developed in Asian countries, e.g., Japan and Korea. The prenatal management of SB fetuses differed depending upon the region, i.e., more support for aggressive and intensive treatment in Asia than in Europe and North America. The regional and racial difference in management of and care for spina bifida fetuses will be discussed further based on the data analysis. PMID- 12905005 TI - Locomotion energetics and gait characteristics of a rat-kangaroo, Bettongia penicillata, have some kangaroo-like features. AB - The locomotory characteristics of kangaroos and wallabies are unusual, with both energetic costs and gait parameters differing from those of quadrupedal running mammals. The kangaroos and wallabies have an evolutionary history of only around 5 million years; their closest relatives, the rat-kangaroos, have a fossil record of more than 26 million years. We examined the locomotory characteristics of a rat-kangaroo, Bettongia penicillata. Locomotory energetics and gait parameters were obtained from animals exercising on a motorised treadmill at speeds from 0.6 m s(-1) to 6.2 m s(-1). Aerobic metabolic costs increased as hopping speed increased, but were significantly different from the costs for a running quadruped; at the fastest speed, the cost of hopping was 50% of the cost of running. Therefore B. penicillata can travel much faster than quadrupedal runners at similar levels of aerobic output. The maximum aerobic output of B. penicillata was 17 times its basal metabolism. Increases in speed during hopping were achieved through increases in stride length, with stride frequency remaining constant. We suggest that these unusual locomotory characteristics are a conservative feature among the hopping marsupials, with an evolutionary history of 20-30 million years. PMID- 12905006 TI - In vitro protein synthesis capacities in a cold stenothermal and a temperate eurythermal pectinid. AB - The translational system was isolated from the gills of the Antarctic scallop Adamussium colbecki (Smith) and the European scallop Aequipecten opercularis (Linnaeus) for in vitro protein synthesis capacities microg protein mg FW(-1) day(-1)) and the translational capacities of RNA (k(RNA in vitro) mg protein mg RNA(-1) day(-1)). In vitro protein synthesis capacity in the cold-adapted pectinid at 0 degrees C was similar to the one found in the temperate scallop at 25 degrees C. These findings might reflect cold compensated rates in Adamussium colbecki, partly explainable by high tissue levels of RNA. Cold-compensated in vitro protein synthesis capacities may further result from increments in the translational capacity of RNA. The thermal sensitivity of the translation machinery was slightly different in the two species, with significantly lower levels of Arrhenius activation energies E(a) and Q(10) in Adamussium colbecki in the temperature range 0-15 degrees C. Reduced protein synthesis and translational capacities were found in vitro in gills of long-term aquarium-maintained Adamussium colbecki and were accounted for by a loss of protein synthesis machinery, i.e. a reduction in RNA levels, as well as a decrease in the amount of protein synthesized per milligram of RNA (RNA translational capacity, k(RNA in vitro)). Such changes may involve food uptake or mirror metabolic depression strategies, like those occurring during winter. Consequences of high in vitro RNA translational capacities found in the permanently cold-adapted species are discussed in the context of seasonal food availability and growth rates at high latitudes. PMID- 12905007 TI - Radiation for prostate cancer issue. PMID- 12905008 TI - High dose rate brachytherapy in the treatment of prostate cancer. AB - The optimal treatment of patients with localized prostate cancer remains controversial. Significant clinical data are available, however, demonstrating that patients treated with radiation therapy (RT) have a significantly better outcome as the dose to the gland is increased. What remains debatable, however, is how to best deliver these higher doses of RT without significantly increasing normal tissue toxicities. Conformal high dose rate brachytherapy (C-HDR BT) represents an alternative means of precise dose delivery that offers similar tumoricidal effects as three-dimensional (3D) conformal external beam radiotherapy (EBRT) or permanent interstitial prostate seed implants with potential additional advantages. Since C-HDR BT consists of temporarily placing afterloading needles or catheters directly into the prostate gland under real time ultrasound guidance, a steep dose gradient between the prostate and adjacent normal tissues can be generated that is minimally affected by organ motion and edema or treatment setup uncertainties. The ability to control the amount of time the single HDR radioactive source "dwells" at each position along the length of each brachytherapy catheter further enhances the conformity of the dose. In addition, recent radiobiological data on prostate cancer treatment suggest that C HDR BT should produce tumor control and late normal tissue side effects that are at least as good as achieved with conventional fractionation, with the additional possibility that acute side effects might be reduced. Published data from several groups performing C-HDR BT as boosts in patients with locally advanced disease have supported these assumptions. Combined with the physical advantages discussed above, C-HDR BT should provide similar tumor control as 3D conformal EBRT with the added advantages of reduced treatment times, less acute toxicity, and no additional technological requirements to account and correct for treatment setup uncertainties and organ motion. Due to the success of C-HDR BT as boost treatment in locally advanced disease, this form of radiation treatment has recently been applied to low-risk prostate cancer patients as an alternative brachytherapy technique to permanent interstitial seed implantation. Advantages in this setting include an improved ability to define and deliver the prescribed dose, a significantly shortened treatment schedule compared to 3D conformal EBRT, and the fact that patients are not radioactive after implantation. PMID- 12905009 TI - Induction of apoptosis in mouse liver adenoma and carcinoma in vivo by transforming growth factor-beta1. AB - PURPOSE: In the liver, transforming growth factor beta-1 (TGF-beta1) constitutes a major negative growth regulating factor involved in the control of cell numbers; failure of this control mechanism has been associated with the development of liver cancer. Since no reports on the in vivo effects of exogenously administered TGF-beta1 on apoptosis in liver tumors have been published yet, we studied hepatocyte sensitivity to the proapoptotic action of TGF-beta1 in stages of chemically induced mouse liver carcinogenesis. METHODS: Mouse liver carcinogenesis was initiated by a single dose of N nitrosodiethylamine (NDEA, 90 mg/kg b.w., i.p.) to 5-week-old B6C3F1 mice. After 2 weeks, mice received either standard diet or a diet containing phenobarbital (PB, 90 mg/kg b.w) for 85 weeks. Four hours before being killed mice received a single dose of TGF-beta1 (56 microg or 200 microg TGF-beta1/kg of b.w., injected into the tail vein). Quantitative histological analysis of mitosis and apoptosis in normal liver tissue (NL), putative preneoplastic foci (PPF), hepatocellular adenoma (HCA), and hepatocellular carcinoma (HCC) was performed on H&E-stained liver sections. RESULTS: In NDEA and NDEA + PB-treated mice, NL exhibited a very low incidence of apoptosis and mitosis, which increased in HCA and HCC. In the lesions apoptoses ranged between 0.03 and 0.6%. Two hundred micrograms of TGF beta1/kg stimulated apoptoses in NL as well as in neoplastic lesions (significant increase in NL, HCA, and HCC); the most pronounced proapoptotic action of TGF-ss1 was observed in lesions of NDEA+PB pretreated mice (about 1.7%). Fifty-six microg TGF-beta1/kg had no detectable effect on apoptosis. CONCLUSION: These observations indicate that during chemically induced liver carcinogenesis in B6C3F1 mice basal rates of apoptoses in adenoma and carcinoma are higher than in normal liver and can be further increased by a proapoptotic cytokine. PMID- 12905010 TI - Dendritic cell-based tumor vaccine for cervical cancer I: in vitro stimulation with recombinant protein-pulsed dendritic cells induces specific T cells to HPV16 E7 or HPV18 E7. AB - PURPOSE: Human papillomavirus (HPV) type 16 and 18 are the most prevalent genotypes in cervical cancers. The viral oncoproteins E6 and E7 are considered to be tumor-specific targets for immunotherapy. HPV E7 antigen-loaded dendritic cells (DC) were evaluated as cellular tumor vaccine. METHODS: Autologous monocyte derived DCs loaded with recombinant HPV16 or HPV18 E7 oncoprotein were used to induce in vitro a specific T cell response. Specificities of activated T cells were determined. RESULTS: E7-specific T cells could be identified in 18/20 T cell lines from healthy blood donors. CD4(+) T cell responses (13/16) were found by proliferation assay. CD8(+) CTLs (12/18) were detectable by interferon-gamma (IFN gamma) ELISpot analysis. Seven donors reacted in both assays and only 2/20 T cell lines did not react in any assay. Thus, specific T cells could be activated in >80% of healthy individuals. T cell lines from suitable donors were specific for HLA-A*0201-restricted epitopes. Furthermore, HPV E7 antigen-loaded DC stimulated specific responses in freshly isolated tumor infiltrating lymphocyte (TIL) populations of cervical cancer patients. CONCLUSION: Autologous dendritic cells loaded with HPV E7 protein can induce T cell responses in healthy individuals by in vitro stimulation and evoke responses in TIL from cervical cancer biopsies. Since there are no limitations with respect to specific HLA-haplotypes, these findings may be a basis for the development of a therapeutic protein-based DC tumor vaccine against cervical cancer for HPV16- and HPV18-positive patients. PMID- 12905012 TI - An unspecific immunostimulating agent and juvenile dermatomyositis: enhanced T cell proliferation and reverse immunosuppression as a severe adverse drug reaction. PMID- 12905011 TI - The late consequences of anthracycline treatment on left ventricular function after treatment for childhood cancer. AB - The purpose of this study was to determine the incidence of changes in left ventricular function in patients in long-term remission after treatment with anthracyclines for a childhood malignancy. The authors examined 155 patients in disease remission who underwent treatment protocols utilising anthracyclines in childhood. The group comprised 90 males and 65 females aged 15+/-4.9 years (range 5-29 years, median 15 years). The age at the time of diagnosis and start of treatment was 8.6+/-4.9 years (range 1-18 years, median 8 years). The time of follow-up was 7.3+/-4 years (range 1-21 years, median 6.3 years). The patients were given a cumulative dose of doxorubicin or daunorubicin of 250+/-131 mg/m2 (range 50-1200 mg/m2, median 240 mg/m2). The values of ejection fraction below 55% and fractional shortening below 30% assessed by means of echocardiography were considered as pathological. The control group consisted of 41 volunteers. Pathological values of fractional shortening were found in 12 patients (8%). Only one patient (0.64%) showed the development of heart failure due to cardiomyopathy. The group of the patients after chemotherapy revealed significantly worse values of left ventricular endsystolic wall stress, mean velocity of circumferential fibre shortening, Tei index, and isovolumic relaxation period in comparison with the control group. We found a correlation between the given cumulative dose of anthracyclines and indicators of systolic function of the left ventricle, but not a relation to the time indicators (age at diagnosis, time of follow-up). CONCLUSION: in the mean period of 6 years after chemotherapy, subclinical cardiotoxicity was found in 11 patients (7%) and cardiomyopathy with heart failure in one patient. Further indicators of subclinical damage are elevation of afterload (end-systolic stress), impaired relaxation and increased value of the Doppler index of global left ventricular function. Further monitoring and evaluation of the relevant subclinical abnormalities over a longer period of time are needed. PMID- 12905013 TI - Paroxetine withdrawal in a neonate with historical and laboratory confirmation. PMID- 12905014 TI - Congenital disorder of glycosylation type 1a in a macrosomic 16-month-old boy with an atypical phenotype and homozygosity of the N216I mutation. AB - We report on a 16-month-old boy with congenital disorder of glycosylation type 1a (CDG-1a; OMIM 212065) showing an atypical phenotype. Whereas failure to thrive is known to be a prominent feature in this condition, our patient developed postnatal macrosomia with increase of weight, length and occipitofrontal circumference (OFC) above the 95th percentile within his 1st year of life. Thereafter, weight and length were close to the 90th and OFC at the 50th percentiles. In contrast to other CDG-1a patients, the child did not have abnormal fat pads or inverted nipples; but unusual eyebrows were present. CDG-1a was confirmed by isoelectric focusing of serum transferrin and measurement of phosphomannomutase activity in leucocytes and cultured fibroblasts (residual activity <5% of controls). Mutation analysis of the phosphomannomutase 2 gene (PMM2) revealed homozygosity for a 647A>T (N216I) mutation in our patient and heterozygosity in his consanguineous parents. CONCLUSION: This is the first report of macrosomia and of homozygosity for the 647A>T (N216I) mutation in a patient with congenital disorder of glycosylation type 1a which may allow further phenotype/genotype comparisons. PMID- 12905015 TI - A mitochondrial DNA mutation (A3243G mtDNA) in a family with cyclic vomiting. PMID- 12905016 TI - Immunogenicity of recombinant BCG vaccine strains overexpressing components of the antigen 85 complex of Mycobacterium tuberculosis. AB - The components of antigen 85 complex of Mycobacterium tuberculosis (Ag 85A, Ag 85B and Ag 85C), due to their immunodominant and secretory nature, represent promising protective antigen candidates and have been used in numerous vaccine preparations. We have used recombinant Bacille Calmette Guerin (BCG) strains overexpressing Ag 85A and Ag 85C to immunize BALB/c mice to investigate the immunogenicity of these strains. Mice immunized with recombinant BCG strains exhibited an increased humoral immune response when compared to mice immunized with wild-type BCG. The recombinant BCG strain overexpressing Ag 85A also induced an increased Th1-like response, characterized by elevated levels of IFN-gamma in antigen stimulated splenocyte cultures and a strong IgG2a antibody response, when compared to wild-type BCG. Immunization with recombinant BCG strain overexpressing Ag 85C, on the other hand did not elicit increased IFN-gamma secretion on restimulation of splenocytes in vitro. PMID- 12905017 TI - Expression and function of bone morphogenetic proteins in the development of the embryonic endocardial cushions. AB - Bone morphogenetic proteins (BMPs) are considered to be significant factors in the morphogenesis of the endocardial cushions of the developing embryonic heart. Previous studies have suggested that they are involved in the epithelial mesenchymal transformation and migration of the cells forming the cushions, or in triggering an apoptotic cascade in a sub-population of cushion cells. We investigated the expression and function of BMP2 and BMP4 proteins in the developing heart of the chick and mouse embryos. In the chick, by immunocytochemistry, we find expression of BMP2 protein in the endocardial cushions of the outflow tract (OT) and atrio-ventricular (AV) regions at embryonic days (ED) 5-6, as well as in adjacent myocardial layers. Immunoblotting indicated that such expression persisted through ED 4-7, but peaked at ED4-5 in the OT and 5-6 in the AV cushions. This temporal sequence correlated with the peaks of apoptotic cell death found previously in the OT and AV cushions of the chick embryo. At equivalent stages in mouse, no such expression of BMP2 was found in the cushions, although expression was detected in adjacent myocardial layers. In the case of BMP4, in both chick and mouse, expression was found only in the myocardia and not in the cushions. Furthermore, BMP-specific receptors were found in the cushions, but not the myocardia, in both the AV and OT regions of the chick embryo. These results provide circumstantial evidence to support the contention that BMPs, originating from the myocardium, could be significant in the induction of apoptosis in chick embryo cushion cells, and confirms that there is species-specific variation in the expression pattern of BMP proteins, as had been predicted from previous studies of mRNA expression. Culture media conditioned by the growth of tissues from various regions of the developing heart were tested for their ability to induce apoptosis in cushion cells in culture. It was found that medium derived from the myocardia induced significant levels of cell death in the cushion cells, and that BMP4 could be detected in such media; however, retroviral over-expression of constitutively active (CA) and dominant negative (DN) isoforms of BMP-specific receptors 1A and 1B (BMPR-1A and BMPR-1B) in cultured cells of the AV cushions did not alter levels of apoptosis or cell proliferation. Similar over-expression in cultured endocardial cells resulted in a significant change in cell shape, from endothelial to fibroblastic, with BMPR 1A CA and BMPR-1B DN, suggesting an influence of these receptors on cell transformation and/or cell migration. Taken together, these results provide support for the contention that BMP2 and BMP4 are important factors in the phenotypic transformational events involved in the morphogenesis of the chick embryo endocardial cushions, and could be involved in the induction of apoptosis in the cushion cells. PMID- 12905018 TI - Unique follicular carcinoma of the thyroid gland with extracellular deposition of amorphous globular structures mimicking an adenoid cystic pattern. PMID- 12905019 TI - Msx genes are expressed in the carapacial ridge of turtle shell: a study of the European pond turtle, Emys orbicularis. AB - The turtle shell forms by extensive ossification of dermis ventrally and dorsally. The carapacial ridge (CR) controls early dorsal shell formation and is thought to play a similar role in shell growth as the apical ectodermal ridge during limb development. However, the molecular mechanisms underlying carapace development are still unknown. Msx genes are involved in the development of limb mesenchyme and of various skeletal structures. In particular, precocious Msx expression is recorded in skeletal precursors that develop close to the ectoderm, such as vertebral spinous processes or skull. Here, we have studied the embryonic expression of Msx genes in the European pond turtle, Emys orbicularis. The overall Msx expression in head, limb, and trunk is similar to what is observed in other vertebrates. We have focused on the CR area and pre-skeletal shell condensations. The CR expresses Msx genes transiently, in a pattern similar to that of fgf10. In the future carapace domain, the dermis located dorsal to the spinal cord expresses Msx genes, as in other vertebrates, but we did not see expansion of this expression in the dermis located more laterally, on top of the dermomyotomes. In the ventral plastron, although the dermal osseous condensations form in the embryonic Msx-positive somatopleura, we did not observe enhanced Msx expression around these elements. These observations may indicate that common mechanisms participate in limb bud and CR early development, but that pre differentiation steps differ between shell and other skeletal structures and involve other gene activities than that of Msx genes. PMID- 12905020 TI - A developmental gradient in the mechanism of K+ uptake during light-stimulated leaf growth in Nicotiana tabacum L. AB - Light causes growth of dicotyledonous leaves by stimulating proton efflux, cell wall acidification and loosening, and solute accumulation for turgor maintenance. For cells still undergoing cell division at the base of expanding tobacco ( Nicotiana tabacum L. cv. Xanthi) leaves, light-stimulated growth depends on K+ uptake, and is inhibited by the potassium channel blocker tetraethylammonium (TEA). The generality of this mechanism has been tested by comparing the effect of light on the growth-associated physiology of dividing and expanding cells in the base with cells at the tip growing by cell expansion only. The magnitude of the light-induced growth response of excised leaf discs is greatest at the leaf base and declines as cells mature. Basal tissue is more sensitive to exogenous potassium, which enhances light-stimulated growth at <1 mM, whereas tip tissue requires higher levels (>10 mM). Growth is inhibited by TEA similarly in tip and base. However, light-stimulated K+ uptake and proton efflux respond differently to TEA in tip and basal tissue. In basal tissue, TEA reduces light-stimulated K+ uptake by 60% and inhibits light-stimulated proton efflux. These results agree with those presented by M. Claussen et al. (1997, Planta 201:227-234) showing that auxin-stimulated H+ pump activity and growth in coleoptiles require K+ uptake as an electrical counterbalance to H+ efflux. In contrast, in tip tissue, TEA inhibits light-stimulated K+ uptake by only 17% and does not inhibit proton efflux. Our results suggest that the basipetal gradient in the effect of TEA on light-regulated growth physiology can be explained by TEA effects on K+ uptake: TEA inhibits light-stimulated H+ pump activity, wall acidification and membrane hyperpolarization only in cells dependent on TEA-sensitive channels for light stimulated K+ uptake. Further, our data suggest that younger, basal tissue is dependent on TEA-sensitive, sucrose-stimulatable channels for light-stimulated K+ uptake whereas older, tip tissue is able to use an additional, TEA-insensitive K+ transporter to mediate light-stimulated K+ uptake. PMID- 12905021 TI - Subcellular localization of two types of ferrochelatase in cucumber. AB - It is widely believed that ferrochelatase (protoheme ferrolyase, EC 4.99.1.1), which catalyzes the insertion of ferrous ion into protoporphyrin IX to form protoheme, exists in both plastids and mitochondria of higher plants. By in vitro import assay with isolated pea (Pisum sativum L.) organelles, it has been proposed that one of two isoforms of ferrochelatase (type 1) is dual-targeted into both plastids and mitochondria, and functions for heme biosynthesis in the both organelles. Recently, however, mitochondrial targeting of ferrochelatase is being disputed since pea mitochondria appeared to accept a variety of chloroplast proteins including the type-1 ferrochelatase of Arabidopsis thaliana (L.) Heynh. To clarify the precise subcellular localization of ferrochelatase in higher plants, here we investigated the subcellular localization of two types of ferrochelatase (CsFeC1 and CsFeC2) in cucumber (Cucumis sativus L.). In cotyledons, a significant level of specific ferrochelatase activity was detected in thylakoid membranes, but only a trace level of activity was detectable in mitochondria. Western blot analysis with specific antibodies showed that anti CsFeC2 antiserum cross-reacted with plastids in photosynthetic and non photosynthetic tissues. Anti-CsFeC1 did not cross-react with mitochondria, but CsFeC1 was clearly detectable in plastids from non-photosynthetic tissues. In situ transient-expression assays using green fluorescent protein demonstrated that, as well as CsFeC2, the N-terminal transit peptide of CsFeC1 targeted the fusion protein solely into plastids, but not into mitochondria. These results demonstrated that in cucumber both CsFeC1 and CsFeC2 are solely targeted into plastids, but not into mitochondria. Screening of a cucumber genomic or cDNA library did not allow any other ferrochelatase homologous gene to be isolated. The data presented here imply the reconsideration of mitochondrial heme biosynthesis in higher plants. PMID- 12905022 TI - Temperature-induced greening of Chlorella vulgaris. The role of the cellular energy balance and zeaxanthin-dependent nonphotochemical quenching. AB - When cells of the green alga Chlorella vulgaris Beij. are transferred from growth at 5 degrees C and an irradiance of 150 micromol photons m(-2) s(-1) to 27 degrees C and the same irradiance, they undergo what is normally considered a high-light to low-light phenotypic change. This involves a 3-fold increase in cellular chlorophyll content with a concomitant increase in light-harvesting complex polypeptide levels. This process appears to occur in response to the cellular capacity to utilize the products of photosynthesis, with the redox state of the plastoquinone pool sensing the cellular energy balance. The phenotypic adjustment can be enhanced or blocked using chemical inhibitors that modulate the redox state of the plastoquinone pool. The functional changes in the photosynthetic apparatus that occurred during the high-light to low-light acclimation were examined with special consideration paid to the paradox that 3 (3,4-dichlorophenyl)-1,1-dimethylurea (DCMU)-treated cells, with non-functional photosystem II (PSII), accumulate light-harvesting polypeptides. At the structural and basic functional levels, the light-harvesting complex of the cells treated with DCMU was indistinguishable from that of the untreated, control cells. To examine how PSII was protected in the DCMU-treated cells, we measured the content of xanthophyll-cycle pigments. It appeared that a zeaxanthin dependent nonphotochemical quenching process was involved in PSII protection during greening in the presence of DCMU. Metabolic inhibitors of mitochondrial respiration were used to examine how the change in cellular energy balance regulates the greening process. Apparently, the mitochondrion acts to supply energy to the chloroplast during greening, and inhibition of mitochondrial respiration diminishes chlorophyll accumulation apparently through an increase in the redox state of the plastoquinone pool. PMID- 12905023 TI - Molecular cloning of an arabidopsis homologue of GCN2, a protein kinase involved in co-ordinated response to amino acid starvation. AB - DNA homologous to the yeast ( Saccharomyces cerevisiae) protein kinase gene, GCN2, was amplified from arabidopsis [ Arabidopsis thaliana (L.) Heynh.] RNA and given the name AtGCN2. The AtGCN2 peptide sequence included adjacent protein kinase and histidyl tRNA synthetase-like domains and showed 45% sequence identity with the GCN2 peptide sequence in the protein kinase domain. AtGCN2 transcripts were detectable in RNA from roots, leaves, stems, buds, flowers, siliques and seedlings. GCN2 is required for yeast cells to respond to amino acid starvation. Expression of AtGCN2 in yeast gcn2 mutants complemented the mutation, enabling growth in the presence of sulfometuron methyl, an inhibitor of branched-chain amino acid biosynthesis, and 3-aminotriazole, an inhibitor of histidine biosynthesis. PMID- 12905024 TI - Gravity-induced modification of auxin transport and distribution for peg formation in cucumber seedlings: possible roles for CS-AUX1 and CS-PIN1. AB - Cucurbit seedlings potentially develop a peg on each side of the transition zone between the hypocotyl and root. Seedlings grown in a horizontal position suppress the development of the peg on the upper side of the transition zone in response to gravity. It is suggested that this suppression occurs due to a reduction in auxin levels to below the threshold value. We show in this study that the free indole-3-acetic acid (IAA) content is low, while IAA conjugates are significantly more abundant in the upper side of the transition zone of gravistimulated seedlings, compared to the lower side. A transient increase in mRNA of the auxin inducible gene, CS-IAA1, was observed in the excised transition zone. The result suggests that the transition zone is a source of auxin. Cucumber seedlings treated with auxin-transport inhibitors exhibited agravitropic growth and developed a peg on each side of the transition zone. Auxin-transport inhibitors additionally caused an increase in CS-IAA1 mRNA accumulation at the transition zone, indicating a rise in intracellular auxin concentrations due to a block of auxin efflux. To study the involvement of the auxin transport system in peg formation, we isolated the cDNAs of a putative auxin influx carrier, CS-AUX1, and putative efflux carrier, CS-PIN1, from cucumber (Cucumis sativus L.) plants. Both genes (CS-AUX1 in particular) were auxin-inducible. Accumulation of CS-AUX1 and CS-PIN1 mRNAs was observed in vascular tissue, cortex and epidermis of the transition zone. A reduced level of CS-AUX1 mRNA was observed in the upper side of the gravistimulated transition zone, compared with the lower side. It is therefore possible that a balance in the activities of auxin influx and efflux carriers controls intracellular auxin concentration at the transition zone, which results in lateral placement of a peg in cucumber seedlings. PMID- 12905026 TI - Photosynthetic CO2 uptake in seedlings of two tropical tree species exposed to oscillating elevated concentrations of CO2. AB - Do short-term fluctuations in CO2 concentrations at elevated CO2 levels affect net CO2 uptake rates of plants? When exposed to 600 microl CO2 l(-1), net CO2 uptake rates in shoots or leaves of seedlings of two tropical C3 tree species, teak (Tectona grandis L. f.) and barrigon [Pseudobombax septenatum (Jacq.) Dug.], increased by 28 and 52% respectively. In the presence of oscillations with half cycles of 20 s, amplitude of ca. 170 microl CO2 l(-1) and mean of 600 microl CO2 l(-1), the stimulation in net CO2 uptake by the two species was reduced to 19 and 36%, respectively, i.e. the CO2 stimulation in photosynthesis associated with a change in exposure from 370 to 600 microl CO2 l(-1) was reduced by a third in both species. Similar reductions in CO2-stimulated net CO2 uptake were observed in T. grandis exposed to 40-s oscillations. Rates of CO2 efflux in the dark by whole shoots of T. grandis decreased by 4.8% upon exposure of plants grown at 370 microl CO2 l(-1) to 600 microl CO2 l(-1). The potential implications of the observations on CO2 oscillations and dark respiration are discussed in the context of free-air CO2 enrichment (FACE) systems in which short-term fluctuations of CO2 concentration are a common feature. PMID- 12905025 TI - Alteration of floral organ identity in rice through ectopic expression of OsMADS16. AB - We used a transgenic approach and yeast two-hybrid experiments to study the role of the rice ( Oryza sativa L.) B-function MADS-box gene, OsMADS16. Transgenic rice plants were generated that ectopically expressed OsMADS16 under the control of the maize ( Zea mays L.) ubiquitin1 promoter. Microscopic observations revealed that the innermost-whorl carpels had been replaced by stamen-like organs, which resembled the flowers of the previously described Arabidopsis thaliana (L.) Heynh. mutation superman as well as those ectopically expressing the AP3 gene. These results indicate that expression of OsMADS16 in the innermost whorl induces stamen development. Occasionally, carpels had completely disappeared. In addition, ectopic expression of OsMADS16 enhanced expression of OsMADS4, another B-function gene, causing superman phenotypes. In the yeast two hybrid system, OsMADS16 did not form a homodimer but, rather, the protein interacted with OsMADS4. OsMADS16 also interacted with OsMADS6 and OSMADS8, both of which are homologous to SEPALLATA proteins required for the proper function of class-B and class-C genes in Arabidopsis. Based on the gene expression pattern and our yeast two-hybrid data, we discuss a quartet model of MADS-domain protein interactions in the lodicule and stamen whorls of rice florets. PMID- 12905027 TI - Localization and functional characterization of metal-binding sites in phytochelatin synthases. AB - Metal-binding domains consisting of short, contiguous stretches of amino acids are found in many proteins mediating the transport, buffering, trafficking or detoxification of metal ions. Phytochelatin synthases are metal-activated enzymes that function in the detoxification of Cd(2+) and other toxic metal and metalloid ions. In order to localize Cd(2+)-binding sites, peptide libraries of two diverse phytochelatin synthases were synthesized and incubated with (109)Cd(2+). Distinct binding sites and binding motifs could be localized based on the patterns of Cd(2+)-binding. The number of binding sites was consistent with previous findings for recombinant protein. Positions of binding sites appeared to be conserved even among diverse phytochelatin synthases. Mutant peptide analysis was used to assess the contribution of exemplary amino acids to binding. Several binding motifs contain cysteines or glutamates. For cysteines a strong correlation was found between binding activity and degree of conservation among known phytochelatin synthases. These findings indicate the suitability of peptide scanning for the identification of metal-binding sites. The functional role of several cysteines was investigated by expression of hemagglutinin-tagged phytochelatin synthases in phytochelatin synthase-deficient, Cd(2+)-hypersensitive Schizosaccharomyces pombe cells. The data are consistent with a model suggesting functionally essential metal-binding activation sites in the N-terminal catalytic part of phytochelatin synthases and additional binding sites at the C-terminus not essential for activity. PMID- 12905028 TI - The proton oligopeptide cotransporter family SLC15 in physiology and pharmacology. AB - Mammalian members of the SLC15 family are electrogenic transporters that utilize the proton-motive force for uphill transport of short chain peptides and peptido mimetics into a variety of cells. The prototype transporters of this family are PEPT1 (SLC15A1) and PEPT2 (SLC15A2), which mediate the uptake of peptide substrates into intestinal and renal epithelial cells. More recently, other sites of functional expression of the two proteins have been identified such as bile duct epithelium (PEPT1), glia cells and epithelia of the choroid plexus, lung and mammary gland (PEPT2). Both proteins can transport essentially every possible di- and tripeptide regardless of the substrate's net charge, but operate stereoselectively. Based on peptide-like structures, various drugs and prodrugs are transported as well, allowing efficient intestinal absorption of the compounds via PEPT1. In kidney tubules both peptide transporters can mediate the renal reabsorption of the filtered compounds thus affecting their pharmacokinetics. Recently, two new peptide transporters, PHT1 (SLC15A4) and PHT2 (SLC15A3), were identified in mammals. They possess an overall amino acid identity with the PEPT-series of 20% to 25%. PHT1 and PHT2 were shown to transport free histidine and certain di- and tripeptides, but it is not yet clear whether they are located on the plasma membrane or represent lysosomal transporters for the proton-dependent export of histidine and dipeptides from lysosomal protein degradation into the cytosol. PMID- 12905030 TI - A radiolabeled peptide ligand of the hERG channel, [125I]-BeKm-1. AB - The wild-type scorpion toxin BeKm-1, which selectively blocks human ether-a-go-go related (hERG) channels, was radiolabeled with iodine at tyrosine 11. Both the mono- and di-iodinated derivatives were found to be biologically active. In electrophysiological patch-clamp recordings mono-[127I]-BeKm-1 had a concentration of half-maximal inhibition (IC50 value) of 27 nM, while wild-type BeKm-1 inhibited hERG channels with an IC50 value of 7 nM. Mono-[125I]-BeKm-1 was found to bind in a concentration-dependent manner and with picomolar affinity to hERG channel protein in purified membrane vesicles from transfected human embryonic kidney cells (HEK-293). Under optimized conditions the equilibrium dissociation constant ( Kd) values from saturation and kinetic binding analysis were 13 and 14 pM, respectively. Both the association and dissociation of [(125)I]-BeKm-1 were fast (association rate constant, k(on)=3.6 x 10(7) M(-1)s( 1); dissociation rate constant, k(off)=0.005 s(-1)). Wild-type BeKm-1 displaced binding of [125I]-BeKm-1 with half-maximal inhibitory concentrations of 44 pM. In contrast, competition experiments with a BeKm-1 mutant BeKm-1-K18A, in which the toxin interaction site is disrupted, resulted in a drop in affinity by more than 300-fold as compared to the wild-type toxin. Iberiotoxin and apamin, peptide inhibitors of Ca2+-activated K+-channels, had no effect on [125I]-BeKm-1 binding. Adding the classical rapid delayed rectifier current (IKr) blocker E-4031 reduced binding of [125I]-BeKm-1 to the hERG channel to an IC50 of 7 nM. In autoradiographic studies on rat hearts, binding of [125I]-BeKm-1 was dose dependent and could partially be displaced by the addition of excess amounts of non-radioactive BeKm-1. The density of the radioactive signal was equally distributed in the myocardium of both the ventricle and atria indicating a homogenous expression of hERG channels throughout the heart. PMID- 12905029 TI - Cation channels, cell volume and the death of an erythrocyte. AB - Similar to a variety of nucleated cells, human erythrocytes activate a non selective cation channel upon osmotic cell shrinkage. Further stimuli of channel activation include oxidative stress, energy depletion and extracellular removal of Cl-. The channel is permeable to Ca2+ and opening of the channel increases cytosolic [Ca2+]. Intriguing evidence points to a role of this channel in the elimination of erythrocytes by apoptosis. Ca2+ entering through the cation channel stimulates a scramblase, leading to breakdown of cell membrane phosphatidylserine asymmetry, and stimulates Ca(2+)-sensitive K+ channels, thus leading to KCl loss and (further) cell shrinkage. The breakdown of phosphatidylserine asymmetry is evidenced by annexin binding, a typical feature of apoptotic cells. The effects of osmotic shock, oxidative stress and energy depletion on annexin binding are mimicked by the Ca2+ ionophore ionomycin (1 microM) and blunted in the nominal absence of extracellular Ca2+. Nevertheless, the residual annexin binding points to additional mechanisms involved in the triggering of the scramblase. The exposure of phosphatidylserine at the extracellular face of the cell membrane stimulates phagocytes to engulf the apoptotic erythrocytes. Thus, sustained activation of the cation channels eventually leads to clearance of affected erythrocytes from peripheral blood. Susceptibility to annexin binding is enhanced in several genetic disorders affecting erythrocyte function, such as thalassaemia, sickle-cell disease and glucose-6-phosphate dehydrogenase deficiency. The enhanced vulnerability presumably contributes to the shortened life span of the affected erythrocytes. Beyond their role in the limitation of erythrocyte survival, cation channels may contribute to the triggering of apoptosis in nucleated cells exposed to osmotic shock and/or oxidative stress. PMID- 12905031 TI - Sepiapterin reduces postischemic injury in the rat heart. AB - A reduced availability of tetrahydrobiopterin (BH4), an essential cofactor for NO synthesis, is causally involved in the development of endothelial dysfunction associated with ischemia/reperfusion. We, therefore, investigated the effect of sepiapterin, a substrate for BH4 synthesis, on postischemic injury in myocardial infarction and myocardial stunning. In rats, myocardial stunning was induced by repetitive ischemia (5 x 10-min ligature of the left coronary artery, 5 x 20-min reperfusion) and myocardial infarction by 50-min ligature and 60-min reperfusion. Myocardial blood flow was determined by H2-clearance, regional myocardial function by pulsed Doppler and infarct size by tetrazolium staining. Myeloperoxidase (MPO) activity was measured as a marker of neutrophil extravasation. cGMP was determined in rat serum as an indicator of increased NO synthesis. In animals treated with sepiapterin, regional myocardial function was significantly improved in both myocardial stunning and infarction and infarct size was significantly reduced. MPO activity decreased with sepiapterin treatment in both models. The systemic level of cGMP was reduced both following myocardial stunning and myocardial infarction in the control group. Pretreatment with sepiapterin induced a significant increase of cGMP level at the end of the protocol in both models. Substitution of sepiapterin reduces postischemic injury both in myocardial stunning and infarction apparently by ameliorating the availability of NO, thereby attenuating the activation of neutrophils in ischemia/reperfusion. PMID- 12905033 TI - Mechanisms of Na+-K+-ATPase phosphorylation by PKC in the medullary thick ascending limb of Henle in the rat. AB - Sodium transport correlates with varying Na+-K+-ATPase activity rates along the nephron. Whether differences in Na+-K+-ATPase regulation by protein kinase C dependent phosphorylation are also present has not been tested. We measured the degree of Na+-K+-ATPase alpha1 subunit phosphorylation by the binding of McK-1 antibody to dephosphorylated Ser-23 and Na+-K+-ATPase activity in medullary thick ascending limb of Henle (mTAL) and proximal tubules (PCT). The degree of Na+-K+ ATPase phosphorylation at Ser-23 was lower in mTAL than in PCT (DU 13.43+/-1.99 versus 2.3+/-0.20, respectively, P<0.01) while Na+-K+-ATPase activity was higher in mTAL (3,402+/-83 vs 711+/-158 pmol/mm tubule per hour in PCT, P<0.01). PKC inhibitor RO-318220 10(-6) M decreased phosphorylation in PCT to 125+/-10% ( P<0.05). In mTAL, RO-318220 did not modify the phosphorylation degree or the activity of Na+-K+-ATPase. Both calcineurin inhibitor FK-506 10(-6) M and phorbol 12-myristate 13-acetate (PMA) 10(-6) M increased the degree of Na+-K+-ATPase phosphorylation ( P<0.05) and inhibited Na+-K+-ATPase activity to 657+/-152 and 1,448+/-347 pmol/mm tubule per hour, respectively, in mTAL ( P<0.01). Increase in [Na+]i to 30, 50 and 70 mM resulted in no changes in Na+-K+-ATPase phosphorylation degree or activity in mTAL. Conversely, in PCT increments in [Na+]i were paralleled by decreased phosphorylation (from 120+/-7 to 160+/-15% of controls, P<0.05) and increased Na+-K+-ATPase activity (from 850+/-139 to 1,874+/ 203 pmol/mm tubule per hour, P<0.01). Dopamine (DA) 10(-6) M decreased both Na+ K+-ATPase dephosphorylation to 41.85+/-9.58% ( P<0.05) and Na+-K+-ATPase activity to 2,405+/-176 pmol/mm tubule per hour in mTAL ( P<0.01). RO-318220 reversed DA effects. Data suggest that regulation of the degree of Na+-K+-ATPase alpha1 subunit phosphorylation at Ser-23 and enzyme activity have different mechanisms in mTAL than in PCT, and may help us to understand the physiological heterogeneity of both segments. PMID- 12905032 TI - Blood sampling methodology is crucial for precise measurement of plasma catecholamines concentrations in mice. AB - Epinephrine (E) and norepinephrine (NE) play a major role in regulating metabolism and cardiovascular physiology. Both are secreted in response to stress and their measurement in plasma allows the study of sympathoadrenal function. Several studies investigating sympathoadrenal physiology are conducted using mice. Review of the literature revealed that basal mouse NE and E plasma concentrations range within 4-140 nM depending on the blood sampling method. Such variability doesn't allow study comparison and may conceal catecholamine variations in response to stress. Therefore, our aim was to determine a reliable sampling method to measure mouse plasma catecholamine concentrations. Results showed that arterial catheterization is the most accurate sampling method: E and NE basal levels were similar to those found in humans (1.1+/-0.3 nM and 4.1+/-0.5 nM, respectively). Retro-orbital bleeding led to analogous results. On the contrary, decapitation was stressful for mice and consequently NE and E concentrations were high (24.6+/-2.7 nM and 27.3+/-3.8 nM, respectively). These different bleeding methods were compared in terms of their ability to detect sympathoadrenal system stimulation (cold-pressure test). With catheter and retro orbital samplings the expected increase in NE and E levels was easily perceived. In contrast, with decapitation no significant change in E was detected. In conclusion, arterial-catheter and retro-orbital blood sampling methods appear to be the most accurate procedures for studying the sympathetic nervous system in mice in both unstressed and stressed conditions. PMID- 12905034 TI - Effect of dexamethasone on voltage-gated K+ channels in Jurkat T-lymphocytes. AB - The voltage-gated K+ channel Kv1.3 is an important regulator of lymphocyte function. Activation of lymphocytes is accompanied by stimulation, whereas CD95 induced apoptosis by inhibition, of Kv1.3. The channel serves to maintain cell membrane potential, a prerequisite for signalling through the Ca2+ release activated Ca2+ channel I(CRAC). As glucocorticoids are known to regulate lymphocyte function, the present study addressed the effect of dexamethasone on voltage-gated K+ channels in Jurkat T-lymphocytes. In whole-cell patch-clamp experiments current families evoked by 200-ms potential steps every 15 s from -70 mV to values from -120 to +100 mV revealed the functional expression of voltage gated K+ channels. Pre-treatment of Jurkat T-lymphocytes for 2-3 h with 1 microM dexamethasone led to a significant decrease of voltage-gated K+ currents. Fura-2 fluorescence measurements showed that the readdition of Ca2+ to Ca(2+)-depleted cells led to a rapid increase of cytosolic Ca2+ activity. This increase of Ca2+ activity was blunted by both the K+ channel blocker margatoxin (10 nM) and 24 h pre-treatment with dexamethasone (1 microM). In conclusion, dexamethasone inhibits voltage-gated K+ channels in Jurkat T-lymphocytes, an effect impeding Ca2+ entry through I(CRAC). PMID- 12905035 TI - Nitric oxide in responses of regional kidney perfusion to renal nerve stimulation and renal ischaemia. AB - The mechanisms underlying the relative insensitivity of medullary blood flow (MBF) to sympathetic drive remain unknown. We tested the effects of nitric oxide synthase blockade on regional kidney perfusion responses to electrical renal nerve stimulation (RNS) in pentobarbitone-anaesthetized rabbits. Under control conditions, RNS reduced renal blood flow (RBF), cortical blood flow (CBF) and MBF in a frequency-dependent manner. MBF was always reduced less than CBF or RBF. NG nitro-L-arginine increased mean arterial pressure (31+/-3 mmHg), reduced RBF ( 8+/-1 ml/min) and MBF (-33+/-6 units), enhanced responses to RNS of RBF (from 48+/-6% to -58+/-6% at 2 Hz), CBF (from -38+/-6% to -43+/-4% at 2 Hz) and, particularly at low frequencies, MBF (from +1+/-18% to -32+/-11% at 2 Hz) and potentiated the RBF hyperaemic response following RNS (by 27+/-6% at 4 Hz). When glyceryl trinitrate was co-infused with NG-nitro-L-arginine to restore basal nitrergic tone, responses to RNS and the subsequent hyperaemia were indistinguishable from control. Since resting renovascular tone or perfusion pressure has little impact on MBF responses to RNS, these present observations suggest that NO contributes to the blunted MBF response to RNS. Paradoxically, NO seems to blunt renal hyperaemia following acute RNS-induced ischaemia. PMID- 12905036 TI - Indications and results of liver transplantation for Echinococcus alveolar infection: an overview. AB - BACKGROUND: Alveolar echinococcosis (AE) of the liver, caused by the larval stage of the fox tapeworm Echinococcus multilocularis, has the characteristics of a slow-growing liver cancer. It is one of the rare parasitic diseases for which a parasitolytic drug is not yet available, and AE is lethal in the absence of appropriate therapeutic management. Complete surgical resection of the parasite at an early stage of infection provides favourable prospects for cure, but, due to a long clinical latency, many cases are diagnosed at an advanced stage, so that partial liver resection can be performed in only 35% of patients. Benzimidazole (BZM) treatment is given in inoperable cases but these compounds are only parasitostatic, and lifelong therapy is required. During the past 20 years some centres have considered liver transplantation (LT) for the treatment of incurable AE. METHODS: Our review summarizes the results of this experience based on a series of 47 European patients who received transplants between 1985 and 2002, tries to specify the real place of LT for AE, and underlines the measures that could be undertaken in the future to improve the results. RESULTS: Five-year survival was 71%. Five-year survival without recurrence was 58%. Major technical difficulties related either to previous laparotomies or to the loco regional involvement were observed. The nine early deaths concerned AE patients with a long past-history of symptomatic AE (iterative cholangitis, secondary biliary cirrhosis). Five late deaths were directly related to ongoing AE, located in the brain in three cases, a very rare AE location that was not investigated before LT in these patients. CONCLUSIONS: In general, the pre-LT screening for distant AE metastases appeared insufficient in this series. Heavy immunosuppressive schemes, absence or delayed re-introduction of BZM after LT have clearly played a role in this unfavourable course. This unique experience indicates that, despite major technical difficulties, LT for incurable AE is feasible and could be discussed in very symptomatic cases. Before LT, interventional radiology should be preferred to repeated laparotomies. Pre-LT and post-LT BZM treatment is mandatory. A careful evaluation of possible distant metastases should be done before the decision for LT is made. After LT, the possibility of an ongoing AE must be permanently kept in mind. This could be reduced by lightening the immunosuppressants, carefully following the specific circulating antibodies, and applying a systematic radiological evaluation, not only to the graft but also to the lungs and the brain. PMID- 12905037 TI - A model of desynchronizing deep brain stimulation with a demand-controlled coordinated reset of neural subpopulations. AB - The coordinated reset of neural subpopulations is introduced as an effectively desynchronizing stimulation technique. For this, short sequences of high frequency pulse trains are administered at different sites in a coordinated way. Desynchronization is easily maintained by performing a coordinated reset with demand-controlled timing or by periodically administering resetting high frequency pulse trains of demand-controlled length. Unlike previously developed methods, this novel approach is robust against variations of model parameters and does not require time-consuming calibration. The novel technique is suggested to be used for demand-controlled deep brain stimulation in patients suffering from Parkinson's disease or essential tremor. It might even be applicable to diseases with intermittently emerging synchronized neural oscillations like epilepsy. PMID- 12905038 TI - Synthesis of two-dimensional human walking: a test of the lambda-model. AB - To test the lambda-model version of the equilibrium point hypothesis both for feasibility and validity with respect to the control of terrestrial locomotion, we developed a two-dimensional, eleven-segment musculoskeletal model of the human body including 14 muscle-tendon complexes per leg, three-segment feet, and a physiologically based model of foot-ground interaction. Human walking was synthesized by numerical integration of the coupled muscle-tendon and rigid body dynamics. To this end a control algorithm based on the lambda-model was implemented in the model providing muscle stimulation patterns that guaranteed dynamically stable walking including a balanced trunk. Thus, the timing of the movement is not preset by a central pattern generator but emerges from the interaction of the musculoskeletal system with the control algorithm. The control parameters were found in a trial-and-error approach. The feedforward part of the control scheme consists of just two target configurations each of which is composed of a set of one nominal length per muscle (lambda-model). Variation of gravity reveals that (1) the synthesized walking patterns are close to ballistic walking and (2) this muscularly induced natural walking can only be initiated and maintained in the range between about a tenth and three times earth-bound gravity. Our walking patterns are robust both against parameter variations and shuffling of the swing leg. We discuss our model with respect to gravity scaling, speed control, feedback delay, and the terms "equilibrium point hypothesis" and "central pattern generator." PMID- 12905039 TI - Coherent interaction of dynamical attractors for object-based selective attention. AB - I investigate essential neuronal mechanisms of visual attention based on object based theory and a biased-competition scheme. A neural network model is proposed that consists of two feature networks, FI and FII, and one object network, OJ. The FI and FII networks send feedforward projections to the OJ network and receive feedback projections from the OJ network in a convergent/divergent manner. The OJ network integrates information about sensory features originated from the FI and FII networks into information about objects. I let the feature networks and the object network memorize individual features and objects according to the Hebbian learning rule and create the point attractors corresponding to these features and objects as long-term memories in the network dynamics. When the model tries to attend to objects that are superimposed, the point attractors relevant to the two objects emerge in each network. After a short interval (hundreds of milliseconds), the point attractors relevant to one of the two objects are selected and the other point attractors are completely suppressed. I suggest that coherent interactions of dynamical attractors relevant to the selected object may be the neuronal substrate for object-based selective attention. Bottom-up (FI-to-OJ and FI-to-OJ) neuronal mechanisms separate candidate objects from the background, and top-down (OJ-to-FI and OJ-to-FII) mechanisms resolve object-competition by which one relevant object is selected from candidate objects. PMID- 12905040 TI - Study of neuronal gain in a conductance-based leaky integrate-and-fire neuron model with balanced excitatory and inhibitory synaptic input. AB - Neurons receive a continual stream of excitatory and inhibitory synaptic inputs. A conductance-based neuron model is used to investigate how the balanced component of this input modulates the amplitude of neuronal responses. The output spiking rate is well described by a formula involving three parameters: the mean mu and variance sigma of the membrane potential and the effective membrane time constant tauQ. This expression shows that, for sufficiently small tauQ, the level of balanced excitatory-inhibitory input has a nonlinear modulatory effect on the neuronal gain. PMID- 12905041 TI - A kinematic theory of rapid human movement. Part IV: a formal mathematical proof and new insights. AB - A few years ago a kinematic theory was proposed to study and analyze rapid human movements. The theory relies on a model of a synergy made up of two neuromuscular systems, one agonist and the other antagonist to the movement. Representing these systems with lognormal impulse responses, it is predicted that the velocity profile of a fast movement will be described by a delta-lognormal equation. So far, many studies have been conducted to test and empirically validate the theory. This paper presents an extended mathematical proof of the model. The proof is based on the Central Limit Theorem under the assumption that a law of proportionate effect governs the cumulative time delays of a sequence of dependent subprocesses constituting a neuromuscular system. Furthermore, a detailed interpretation of the parameters of the delta-lognormal equation, in terms of movement time and amplitude, response time and time delays, is discussed, providing new insights into the properties of the model with respect to neuromuscular system activity and movement generation. PMID- 12905042 TI - A model of the feline medial gastrocnemius motoneuron-muscle system subjected to recurrent inhibition. AB - Recurrent inhibition in the mammalian spinal cord is complex, and its functions are not yet well understood. Skeletomotoneurons (alpha-MNs) excite, via recurrent axon collaterals, inhibitory Renshaw cells (RCs), which in turn inhibit alpha-MNs and other neurons. The anatomical and functional structure of the recurrent inhibitory network is nonhomogeneous, and the gain and filtering characteristics of RCs are modulated by inputs circumventing alpha-MNs. This complex organization is likely to play important roles for the discharge and recruitment properties of alpha-MNs. Modeling this system is a way of investigating hypothesized roles for normal functioning including muscle fatigue and different forms of physiological pathological tremor. In this paper, a detailed model including alpha-MNs, RCs, and the muscle fibers innervated by the alpha-MNs is presented. Outlines of the experimental data underlying the model and the modeling philosophy and procedure are presented. Then the behavior of a RC model is compared with experimental data reported in the literature. Model and experimental data agree well for burst responses elicited by synchronous single-pulse activation of different numbers of motor axons. In addition, the static relation between motor-axon activation rate and RC firing rate agree fairly well in model and experiment, and the same applies to the dynamic responses to step changes in motor-axon rate. The ultimate objective is to use this model in probing the role of recurrent inhibition in the control and stability of (isometric) muscular force under normal and altered conditions occurring during fatigue and muscle pain. PMID- 12905043 TI - Muscle synergies during shifts of the center of pressure by standing persons: identification of muscle modes. AB - When a standing person performs a movement such that the center of gravity shifts, the activity of postural muscles adjusts to keep the balance. We assume that such adjustments are controlled using a small set of central variables, while each variable induces changes in the activity of a subgroup of postural muscles. The purpose of this study has been to identify such muscle groups (muscle modes or M-modes) and compare them across tasks and subjects. Four tasks required the subjects to release a load from extended arms leading to a center of pressure (COP) shift prior to the load release. The fifth task required an explicit COP shift by voluntary sway. Electromyographic activity of 11 postural muscles on one side of the body was integrated over a 100-ms interval corresponding to the early stage of the COP shift, and this integrated EMG activity was subjected to a principal component (PC) analysis across multiple repetitions of each task. Three PCs were identified and associated with a "push back M-mode," a "push-forward M-mode," and a "mixed M-mode." Cluster analysis of the PC vectors across tasks and across subjects confirmed the existence of distinctive push-forward and push-back muscle groups. PC vectors were also compared across tasks and across subjects using cosines as a measure of colinearity between pairs of vectors. In general, M-modes were similar across both tasks and subjects. We conclude that shifts of the COP, whether implicit or explicit, are controlled using a small set of central variables associated with changes in the activity of robust subsets of postural muscles. These results can be used for future analysis of muscle synergies associated with postural tasks. PMID- 12905044 TI - Region-specific differences in Achilles tendon cross-sectional area in runners and non-runners. AB - The present study examined the cross-sectional area (CSA) of the Achilles tendon in subjects who repeatedly exposed their tendons to large loads (habitual runners) compared to control subjects (non-runners). Six male habitual runners [36 (7) years, 70.9 (4.4) kg and 1.84 (0.05) m, +/-SD] who had performed distance running (approximately 80 km per week) for the last 5 years were compared to six non-runners [34 (3) years, 81.2 (8.7) kg and 1.81 (0.02) m, +/-SD]. Tendon CSA was obtained from MR images obtained with the ankle in a neutral position (90 degrees ). The most proximal aspect of the tuberosity of calcaneus was used as a landmark to standardize the levels of images: the most distal image (1) was obtained 10 mm above the proximal tuberosity of calcaneus, and the most proximal image (7) was obtained 70 mm above the proximal tuberosity of calcaneus. There was a significant difference in CSA along the length of the tendon both in runners ( P<0.001) and non-runners ( P<0.01). In non-runners and runners the CSA of the most distal part was 51% and 85% greater than the most proximal part of the tendon, respectively. Furthermore, there was a difference in tendon CSA between the groups, such that runners had a greater CSA (36%) than non-runners at the most distal part of the tendon ( P<0.05). The greater CSA in the distal tendon may reflect differences in structural properties along the length of the human Achilles tendon, while the greater CSA in runners compared to non-runners may indicate a region-specific hypertrophy in response to the habitual loading of running. PMID- 12905045 TI - Leptin as a marker of training stress in highly trained male rowers? AB - The purpose of this study was to investigate the potentially important role leptin may play during training monitoring in athletes. Twelve highly trained male rowers underwent a 3-week period of maximally increased training stress followed by a 2-week tapering period. Fasting blood was sampled after a rest day. Subjects also performed a maximal 2000-m rowing ergometer test before and after 3 weeks of heavy training, and after 2 weeks of tapering. Blood samples were obtained before, immediately after and after 30 min of recovery. Leptin concentrations were measured in duplicate by radioimmunoassay. Mean training time was about 100% higher during the heavy training period (17.5 h x week(-1)) compared to the tapering period (8.9 h x week(-1)). The 3-week heavy training period induced a significant reduction ( P<0.05) in the fasting leptin concentration [from 2.5 (0.4) to 1.5 (0.4) ng x ml(-1)]. Fasting plasma leptin was significantly increased by the end of the 2-week tapering period [2.0 (0.4) ng x ml(-1)] but remained significantly lower compared to the pretraining value. Leptin levels were also significantly decreased only after the 2000-m rowing ergometer test performed at the end of the heavy training period. No differences in leptin concentrations were observed after other performance tests compared to their respective baseline values. In addition, fasting leptin concentration was significantly related to the weekly training time ( r=-0.45; P=0.006). In conclusion, it appears that leptin is sensitive to the rapid and pronounced changes in training volume. A greater training time is associated with a lower leptin concentration in highly trained male rowers. It is suggested that it may be possible to direct typical rowing training by monitoring leptin status. PMID- 12905047 TI - Determination of rate-constants as a method to describe passive expiration. AB - To describe the relaxed expiration by a two-compartment model, we introduced a gas/energy transfer between the lung compartment ( V(1)) and a second one ( V(2)). If V(2) were a real volume, the rate-constants (i.e. the flow/volume ratios) of the compartments would describe a real gas-exchange. Alternatively, if a viscoelastic behaviour of the lung or an energy-exchange between compartments was simulated, V(2) would become a "pseudo-volume". We studied nine mechanically ventilated subjects. Changes in volume were transduced by respiratory inductive plethysmography. The rate-constants were assumed (together with the initial volumes of the compartments) as parameters to fit the total volume [ V(1)( t)+ V(2)( t)]. Once the best fitting was performed using these "physiological" parameters, the system was directly identified and the compartments were independently analysed. The time profile of the second compartment showed a maximum that depended on the value of the rate-constants. Appropriate tests confirmed the reliability of our procedure. In conclusion, our analysis demonstrated that the energy/volume of the second compartment may increase at the beginning of expiration and then decrease, showing a maximum, even though the total curve can only be a decreasing one. In other words, the slowing down of the curve representing expiratory volume is due not only to the longer emptying of the second compartment, but also to the interaction between the two compartments. As presently proposed, this interaction can be represented by either a gas exchange between two actual volumes, or a mechanical energy transfer between the lung and the tissue compartment. PMID- 12905046 TI - The effect of reactive oxidant generation in acute exercise-induced proteinuria in trained and untrained rats. AB - Exercise-induced proteinuria is a common consequence of physical activity, although its mechanism is not clear. We investigated whether free radicals generated during exercise play a role in post-exercise proteinuria in sedentary and treadmill-running trained rats, separately. Sedentary and trained rats were randomly divided into four sub-groups: control, antioxidant treatment, exhaustive exercise and an exhaustive exercise plus antioxidant treatment group. Antioxidant therapy was applied by intragastric catheter for 4 weeks with vitamin C (ascorbic acid, 50 mg x kg(-1) x day(-1)) and vitamin E (alpha-tocopherol, 20 mg x kg( 1).day(-1)). Twenty-four-hour urine samples were used for measuring protein levels and protein electrophoresis. Thiobarbituric acid (TBARS) and glutathione (GSH) levels, superoxide dismutase (SOD) and catalase (CAT) activities were assayed in blood and tissues. Increased urinary protein levels and mixed type proteinuria in electrophoresis were identified after exhaustive exercise in sedentary rats. Erythrocyte, kidney and muscle TBARS levels were significantly elevated in this group. Antioxidant treatment prevented the increase in urinary protein levels, TBARS levels and the occurrence of mixed type proteinuria after exhaustive exercise in sedentary rats. Exhaustive exercise in trained rats resulted in elevation of urine protein levels and mixed type proteinuria although kidney TBARS levels were not changed compared to those of the trained controls. Antioxidant therapy in trained and exhausted-trained animals resulted in decreased TBARS levels in the kidney but it did not affect urinary-increased protein levels or electrophoresis in exhausted animals. This findings suggest that the exercise-induced oxidant stress may contribute to post-exercise proteinuria in sedentary rats. However, this mechanism may not be responsible for proteinuria in trained rats. PMID- 12905048 TI - Variability of gait patterns during unconstrained walking assessed by satellite positioning (GPS). AB - It is established that the ratio between step length (SL) and step frequency (SF) is constant over a large range of walking speed. However, few data are available about the spontaneous variability of this ratio during unconstrained outdoor walking, in particular over a sufficient number of steps. The purpose of the present study was to assess the inter- and intra-subject variability of spatio temporal gait characteristics [SL, SF and walk ratio (WR=SL/SF)] while walking at different freely selected speeds. Twelve healthy subjects walked three times along a 100-m athletic track at: (1). a slower than preferred speed, (2). preferred speed and (3). a faster than preferred speed. Two professional GPS receivers providing 3D positions assessed the walking speed and SF with high precision (less than 0.5% error). Intra-subject variability was calculated as the variation among eight consecutive 5-s samples. WR was found to be constant at preferred and fast speeds [0.41 (0.04) m.s and 0.41 (0.05) m.s respectively] but was higher at slow speeds [0.44 (0.05) m.s]. In other words, between slow and preferred speed, the speed increase was mediated more by a change in SF than SL. The intra-subject variability of WR was low under preferred [CV, coefficient of variation = 1.9 (0.6)%] and fast [CV=1.8 (0.5)%] speed conditions, but higher under low speed condition [CV=4.1 (1.5)%]. On the other hand, the inter-subject variability of WR was 11%, 10% and 12% at slow, preferred and fast walking speeds respectively. It is concluded that the GPS method is able to capture basic gait parameters over a short period of time (5 s). A specific gait pattern for slow walking was observed. Furthermore, it seems that the walking patterns in free living conditions exhibit low intra-individual variability, but that there is substantial variability between subjects. PMID- 12905050 TI - Relative contributions of central and peripheral factors to fatigue during a maximal sustained effort. AB - Local muscle fatigue can originate from both peripheral and central factors. The relative contribution of these factors in the course of a fatiguing contraction in 20 healthy subjects was determined. While subjects made a 2-min sustained maximal voluntary contraction (MVC) of the biceps brachii, muscle fibre conduction velocity (MFCV) was determined with surface electromyography (SEMG) as a representation of developing peripheral fatigue. To quantify the amount of peripheral fatigue, the force development following a train of electrical stimuli on the endplate before and after the contraction were compared. To measure force loss caused by central factors, superimposed electrical stimulation was used during the contraction. By two different methods the influence of peripheral fatigue on the superimposed force responses was taken into account. The first method compared the force response with the actual voluntary force, the second which seemed more valid - used an estimation of peripheral fatigue based on linear interpolation between the force responses during rest before and after sustained contraction. During the contraction, voluntary force decreased to 38%. Peripheral fatigue was responsible for the larger part of this decline (89%). The other part, which was calculated as 12%, was caused by an increase of central activation failure. The decline of MFCV indicated that peripheral fatigue increased predominantly during the first half of the contraction and stayed at a constant level during the latter part. In contrast, central fatigue mainly induced a force decrease in the second part of the contraction. The different mechanisms which could be responsible for this change of emphasis from peripheral to central factors are discussed. PMID- 12905049 TI - Effect of order of exercise intensity upon cardiorespiratory, metabolic, and perceptual responses during exercise of mixed intensity. AB - Exercise of mixed intensities can be of benefit in many different ways. However, whether physiological interaction exists between exercises of different intensity is questionable. As such, the primary aim of this study was to examine the effect of order of exercise intensity upon cardiorespiratory, metabolic, and perceptual responses during exercise of mixed intensity. Eight males and four females volunteered to serve as subjects for the study. They were informed of the purpose of the experiment and gave their written consent to participate. Each subject completed a peak oxygen uptake ( VO(2peak)) test and two submaximal exercises of mixed intensity on three separate laboratory visits. During each submaximal exercise trial, subjects performed a 15-min (high intensity) exercise at 70% VO(2peak) that was followed by another 15-min (low intensity) exercise at 50% VO(2peak) (high/low, H/L), or a 15-min exercise at 50% VO(2peak) that was followed by another 15-min exercise at 70% VO(2peak) (low/high, L/H). Oxygen uptake ( VO(2)), respiratory exchange ratio ( R), expired ventilation ( V(E)), heart rate (HR) and ratings of perceived exertion (RPE) were measured every 5 min throughout exercise. Energy expenditure and carbohydrate and fat oxidation were calculated from VO(2) adjusted for substrate metabolism using R and then accumulated for each phase of exercise intensity as well as for the entire exercise session. VO(2) and HR were higher ( P<0.05), while R was lower ( P<0.05) at the lower intensity in H/L than in L/H. V(E) and RPE were lower ( P<0.05) at the higher intensity in H/L than in L/H. While no differences in caloric expenditure and carbohydrate oxidation between the two trials were observed, fat oxidation was higher ( P<0.05) both at the lower intensity and for the entire trial in H/L than in L/H. It appears that during exercise of mixed intensity, placing some periods of moderate intensity exercise prior to a milder one is a more favorable sequence in that it can elicit a greater fat oxidation while being felt less stressful. PMID- 12905052 TI - Adhesion of Porphyromonas gingivalis to cultured pocket epithelium: mono- and multi-layered. AB - Adhesion of bacteria to epithelial cells might be influenced by the degree of cell differentiation, as observed in the multi-layering process of epithelial cells. In the present study, the adhesion of a large group of clinical Porphyromonas gingivalis strains (n=11) to in vitro cultured mono- and multi layers of epithelial cells was examined and compared. The tissue samples originated from 6 patients with chronic adult periodontitis. Porphyromonas gingivalis bacteria adhered more to mono-layers as opposed to the more differentiated multi-layers. Differences between the clinical P. gingivalis strains, however, became obvious only on multi-layers. These partially differentiated cells may also better represent the individual subject variations. Mono-layer cultures, which are simpler to obtain, seem to be less precise. The importance of cell differentiation on bacterial adhesion needs more attention. PMID- 12905051 TI - Service sector and perceived social support at work in Norwegian nurses' aides. AB - OBJECTIVES: Social support at work is associated with reduced risk of illness and sickness absence. Identifying determinants of social support at work is, hence, important. The objective of this study was to determine the relationship between service sector and level of social support at work in nurses' aides. METHODS: A random sample of nurses' aides, organised by the Norwegian Union of Health-and Social Workers, was mailed a questionnaire in 1999. Of 12,000 recipients, 7,478 (62.3%) completed the questionnaire. The 6,234 vocationally active, female respondents comprised the sample of this study. Perceived social support at work was assessed by scales in the standardised questionnaire, the General Nordic Questionnaire for Psychological and Social Factors at Work (QPSNordic). Each service sector was compared with all other sectors. Because of multiple comparisons, the significance level was set at 0.01, and 99% confidence intervals were used. RESULTS: The following associations were found, after adjustments for age and work schedule: high levels of support from the immediate superior were reported more often by aides in the psychiatric sector [odds ratio (OR) 1.72; 99% confidence interval (CI) 1.29-2.29] and community-nurse service (OR 1.22; CI 1.01 1.48), and less often by aides in nursing homes (OR 0.67; CI 0.58-0.77). Encouraging and supportive culture in the work unit was reported more often by aides in the psychiatric sector (OR 1.55; CI 1.13-2.13), and less often by aides in nursing homes (OR 0.79; CI 0.69-0.92). CONCLUSIONS: In nurses' aides, perceived level of social support at work varies with service sector. Differences in organisational structure, personnel resources, and work tasks could explain the results. It is essential that managers in nursing homes put more emphasis on creating a supportive culture in their work units. Reorganisation and training of personnel could be elements of this effort. Higher grants may also be needed. PMID- 12905053 TI - The effect of perioperative corticosteroids on the outcome of microscopic lumbar disc surgery. AB - Eighty adult patients with lumbar disc herniation verified by magnetic resonance imaging (MRI) and clinical findings corresponding to the radiological level underwent microscopic disc removal to evaluate the outcome of perioperatively given corticosteroids in a prospective randomized double-blind study. In the treatment group the patient received 250 mg Solu-Medrol intravenously and 160 mg Depo-Medrol intramuscularly. Before closure of the wound, a free fat transplant soaked in 80 mg Depo-Medrol was placed on the dural sac. In the control group the same procedure was performed, but sodium chloride was given instead of Depo Medrol. All patients underwent a clinical examination before surgery and at 2, 6, 12, 26, 52 and 104 weeks postoperatively, rating their pain with the visual analog scale (VAS) and function with the Disability Rating Index (DRI). The postoperative hospital stay was significantly shorter (P=0.01) in the treatment group (1.7 days) compared to the control group (2.3 days). Time taken to return to full-time work was also significantly shorter in the treatment group (P=0.003). VAS-W (Worst Pain during last week) was significantly lower in the treatment group (P=0.02). Postoperative spondylitis occurred in one patient in the control group and no adverse corticosteroids effect was seen. Our study shows that perioperatively given corticosteroids improve the outcome of microscopic disc surgery in terms of length of hospital stay and time taken to return to full time work. The results also indicate that corticosteroid treatment reduces pain and improves functional outcome. PMID- 12905054 TI - Preoperative radiological and electrophysiological evaluation in 100 adolescent idiopathic scoliosis patients. AB - This is a prospective study of spinal magnetic resonance imaging (MRI), electrophysiological recordings, and neurological examinations of 100 patients admitted for surgery for adolescent idiopathic scoliosis (AIS), which was conducted to assess the prevalence of structural and functional abnormalities within the spinal cord in patients with clinically normal neurologic condition. In all patients the clinical diagnosis and intact neurological condition was ascertained by a spinal orthopedic surgeon. Full-length spinal axis MRI studies (T1/T2 sequences) and somato-sensory evoked potentials of the tibial nerves (tSSEPs) were preoperatively assessed by independent evaluators blinded to the patients' medical histories. Structural spinal cord abnormalities were found in three of 100 AIS patients on MR imaging. In one patient a Chiari malformation type 1 with an accompanying syringomyelia was diagnosed, which required a suboccipital decompression. In the other two patients small thoracic syringomyelias were diagnosed. Abnormalities of spinal cord function were detected in 68% of the 100 patients: tSSEP latencies corrected for body height were increased in 56% of the patients; pathological differences between tSSEPs on the left and right sides were present in 17% (12% in combination with a prolongation of the latency). The findings of this study indicate that MRI and electrophysiological examinations are essential to assess spinal cord abnormalities that are clinically not detectable in AIS patients. Even in patients with intact neurologic condition and clinically typical right-curved thoracic scoliosis, the possibility of intraspinal pathologies should be ruled out by MRI. It is especially important to detect structural pathologies like syringomyelia and Chiari malformation before proceeding with scoliosis surgery, as these conditions are associated with a higher neurological risk during scoliosis surgery. The electrophysiological recordings made in the present study, with the high number of pathological tSSEPs, are indicative of functional abnormalities with a subclinical involvement of the recorded neuronal pathways. The relevance of the latter findings is not yet clear, but pre-operative tSSEP examinations offer the possibility of assessing alterations in spinal cord function that are undetectable by clinical examination. PMID- 12905056 TI - Lack of marital support and poor psychological responses in male cancer patients. AB - A total of 272 men and 252 women with cancer participated in a study of the impact of being married and the presence of spousal support on psychological distress and coping with cancer. All participants underwent a structured interview and completed the Profile of Mood States and the Mental Adjustment to Cancer scale. Multivariate analysis controlling for potentially confounding biomedical and psychosocial variables revealed that unmarried men had significantly higher levels of psychological distress and lower levels of fighting spirit than married men, and that men with spousal support showed higher levels of fighting spirit than men without spousal support. However, no such difference was observed between unmarried and married women or between women with and without spousal support. These findings suggest that being married may play an important role in reducing psychological distress and enhancing fighting spirit of men with cancer, and that being unmarried may be a risk factor for psychological distress and lower fighting spirit for men with cancer. PMID- 12905055 TI - Pain and its treatment in hospitalized patients with metastatic cancer. AB - GOALS: The aim of this prospective study was to assess the quality of pain management hospitalized cancer patients. PATIENTS AND METHODS: In a quantitative and qualitative evaluation from six oncology centers in Italy, all consecutive cancer patients complaining of pain and hospitalized during the same 2 weeks were requested to fill in a McGill pain questionnaire (MPQ), a present pain intensity scale (PPI), and a hospital anxiety and depression acale (HADS), and to answer a questionnaire focused (QF) on the quality of medical and nursing care. The healthcare provider's antalgic prescriptions were assessed by an index of pain management (IPM). MAIN RESULTS: Of 120 patients with pain admitted to oncology divisions (65 men and 52 women; mean age 57 years, range 21-79 years), 117 completed the questionnaires. The quantitative evaluation (PPI) showed a significant pain reduction between admission and discharge pain levels-from 2.65 to 1.50 ( p<0.001). While a significant reduction of anxiety (HADS) was also found-from 10.24 to 9.11 ( p<0.001)-depression did not improve (9.83 and 9.72). The most relevant information from qualitative evaluation (QF) was: in 37.6% of patients, pain level was higher overnight; 47% waited for spontaneous decrease of pain intensity before asking for nurse or physician intervention; 69% asked for nurse help when pain level was really high. The health care response to patients' pain was not completely satisfactory, since analgesic prescription was adequate in 56.52% but inadequate in 43.47%. CONCLUSIONS: Pain control in hospitalized cancer patients is not completely satisfactory. The physician's attitude is to underestimate and undertreat pain, while nurses are not adequately trained for timely intervention despite published guidelines for pain management. The findings of this study support the concern of inadequate knowledge and inappropriate attitudes regarding pain management, even in cancer patients hospitalized in medical oncology divisions. PMID- 12905058 TI - The relationship between patient characteristics and carer psychological status in home palliative cancer care. AB - GOALS: Despite being both providers and intended recipients of care, informal carers in cancer palliative care report high levels of distress and unmet needs. In order to develop supportive care strategies, this analysis aimed to identify which patient characteristics contribute to carer psychological distress and which coping strategies carers employ. PATIENTS AND METHODS: Informal carers attending two home palliative care services gave cross-sectional data regarding patient characteristics and their own psychological status using standardised measures. Multivariate analyses were performed for each dependent carer psychological measure, with patient characteristics as independent variables (adjusted for carer age and gender). MAIN RESULTS: Forty-three carers participated. Greater patient distress was associated with carer anxiety (b value: magnitude of the effect) (b=0.31, p=0.07), and both patient psychological status (b=0.37, p=0.02) and pain (b=0.29, p=0.09) were associated with carer psychological morbidity. Carer burden was associated with patient psychological distress (b=0.35, p=0.03) and pain (b=0.29, p=0.08). Carer avoidance/emotion focused cognitive coping strategies were associated with patient physical function (b=0.34, p=0.04), and cognitive problem-focused coping was associated with patient symptoms (b=0.28, p=0.06) and physical function (b=0.29, p=0.05). CONCLUSIONS: Adequate provision of patient psychological interventions and effective pain education and control are needed in order to improve carers' psychological health. Patient characteristics are associated with apparently opposing forms of carers' coping (i.e. both avoidance and engagement), demonstrating the importance of interventions addressing a range of coping responses. Further research is needed to understand why carers employ problem focused coping in response to symptoms but not to pain. Evidence-based interventions for informal carers are urgently needed but must be delivered in the context of optimal patient pain and symptom control. PMID- 12905059 TI - Sublethal effect of a weak intermittent magnetic field on the development of Xenopus laevis (Daudin) tadpoles. AB - In three repeated experiments with three different litters of Xenopus laevis (Daudin) tadpoles, three cohorts were reared in an aquarium under the "saw-tooth" magnetic field produced by a television set. Their maturation times are compared with those of three corresponding control cohorts grown in an unexposed aquarium. In the exposed aquarium, the magnetic field amplitude was less than 25 microT and the frequency in the extremely-low-frequency and very-low-frequency wavebands. Neither the exposed nor the unexposed cohorts suffered significant mortality and malformations. However, the exposed tadpoles took about 5 days more than the unexposed ones to reach metamorphosis. The differences in mean maturation times between the exposed and control cohorts were extremely significant (P < 0.001). The results show that a biological population can suffer a sublethal effect when exposed to the magnetic field of a TV set for a long time in the course of juvenile life stages, and that this effect can consist of a delay in reaching the adult stage. PMID- 12905057 TI - Probability of mortality of critically ill cancer patients at 72 h of intensive care unit (ICU) management. AB - GOALS: To develop and validate a model for probability of hospital mortality for cancer patients at 72 h of intensive care unit (ICU) management. PATIENTS AND METHODS: This is an inception cohort study performed at four ICUs of academic medical centers in the United States. Defined continuous and categorical variables were collected on consecutive patients with cancer admitted to the ICU. A preliminary model was developed from 827 patients and then validated on an additional 415 patients. Multiple logistic regression modeling was used to develop the models, which were subsequently evaluated for discrimination and calibration. The main outcome measure is in-hospital death. RESULTS: A probability of mortality model, which incorporates ten discrete categorical variables, was developed and validated. All variables were collected at 72 h of ICU care. Variables included evidence of disease progression, performance status before hospitalization, heart rate >100 beats/min, Glasgow coma score 40 mg/dl, and a urine output of <150 ml for any 8 h in the previous 24 h. The p values for the fit of the preliminary and validation models were 0.535 and 0.354 respectively, and the areas under the receiver operating characteristic (ROC) curves were 0.809 and 0.820. CONCLUSIONS: We report a multivariable logistic regression model to estimate the probability of hospital mortality in critically ill cancer patients at 72 h of ICU care. The model is comprised of ten unambiguous and readily available variables. When used in conjunction with clinical judgment, this model should improve discussions about goals of care of these patients. Additional validation in a community hospital setting is warranted. PMID- 12905060 TI - Mycorrhizal colonization mediated by species interactions in arctic tundra. AB - The Alaskan tussock tundra is a strongly nutrient-limited ecosystem, where almost all vascular plant species are mycorrhizal. We established a long-term removal experiment to document effects of arctic plant species on ecto- and ericoid mycorrhizal fungi and to investigate whether species interactions and/or nutrient availability affect mycorrhizal colonization. The treatments applied were removal of Betula nana ( Betulaceae, dominant deciduous shrub species), removal of Ledum palustre ( Ericaceae, dominant evergreen shrub species), control (no removal), and each of these three treatments with the addition of fertilizer. After 3 years of Ledum removal and fertilization, we found that overall ectomycorrhizal colonization in Betula was significantly reduced. Changes in ectomycorrhizal morphotype composition in removal and fertilized treatments were also observed. These results suggest that the effect of Ledum on Betula's mycorrhizal roots is due to sequestration of nutrients by Ledum, leading to reduced nutrient availability in the soil. In contrast, ericoid mycorrhizal colonization was not affected by fertilization, but the removal of Betula and to a lower degree of Ledum resulted in a reduction of ericoid mycorrhizal colonization suggesting a direct effect of these species on ericoid mycorrhizal colonization. Nutrient availability was only higher in fertilized treatments, but caution should be taken with the interpretation of these data as soil microbes may effectively compete with the ion exchange resins for the nutrients released by plant removal in these nutrient-limited soils. PMID- 12905061 TI - Benefit to N2-fixing alder of extending growth period at the cost of leaf nitrogen loss without resorption. AB - This study examines the adaptive role of not resorbing N in N(2)-fixing deciduous trees in terms of their energy balance. The autumnal growth of N(2)-fixing Alnus firma Sieb. et Zucc. (alder) was compared with that of the non-N(2)-fixing Morus bombycis Koizumi (mulberry), which resorbs leaf N. The freezing resistance of leaves of both species was -2 degrees C. Mulberry seedlings lost their photosynthetic ability in mid-October, although the minimum temperature was still above 0 degrees C. Thereafter, their leaves turned yellow and were gradually shed. In contrast, seedlings of the alder maintained their photosynthetic ability until mid-November, when the minimum temperature fell to the freezing resistance limit. Thereafter, their leaves were shed quickly without an autumn tint. The mulberry resorbed 48.9% of leaf N, whereas the alder resorbed hardly any. These results show that, compared with the mulberry tree, the alder extended its growth period for 1 month in return for losing leaf N without resorption. The amount of energy assimilated by the alder in the extended growth period was about six times that required for compensating for the nitrogen loss, if the compensation is dependent only on the tree's own nitrogen fixation. This surplus energy balance has probably allowed N(2)-fixing deciduous trees to evolve their non-N-resorbing trait. PMID- 12905062 TI - Cellular responses to increasing Cd concentrations in the freshwater crab, Potamonautes warreni, harbouring microbial gill infestations. AB - We investigated the uptake, transport, storage and defence mechanisms in the freshwater crab, Potamonautes warreni, harbouring microbial gill infestations and exposed to increasing chronic (0.2, 0.5, 1.0 mg l(-1)) and acute (2.0 mg l(-1)) cadmium (Cd) concentrations under controlled laboratory conditions over a period of 21 days. Transmission electron microscopy and X-ray microanalysis revealed that the microbial gill fauna was eliminated on exposure to 0.2 mg Cd(2+) l(-1) and that Cd became increasingly adsorbed and incorporated into lamellar crystal deposits and permeated the cuticle of the gills of P. warreni. Degeneration of the apical membrane infoldings and vacuolation of epithelial cells occurred concurrently with pinocytosis, endocytosis and pronounced phagocytotic activity in the epithelia and haemal canal of the gills. Elevated Cd exposures (0.5 or 1.0 mg l(-1)) resulted in the swelling and dissociation of mitochondrial outer membranes together with an increase in transport of Cu, Cl and S by haemocytes in the haemal canal to epithelial tissues depleted in these elements. Cd also accumulated in tightly coiled concentric membrane whorls in the haemal canal, whereas the highest concentrations of Cd were found within aggregates of lysosome like bodies in cuticulin-secreting cells of the gill stem. Chronic exposure to Cd induced increased fatigue and mild uncoordinated motor activity. In contrast, at an acute exposure of 2.0 mg l(-1) over 48 h, P. warreni showed a time-specific rapid loss of motor function, although only mild cellular lesions occurred in the gill tissues. The significance of cellular changes in the gill epithelia and altered motor activity of P. warreni with increased waterborne Cd are discussed as potential biomarker responses in monitoring aquatic pollution. PMID- 12905063 TI - Expression patterns of odorant-binding proteins in antennae of the moth Manduca sexta. AB - Monoclonal antibodies (MAbs) were generated to six recombinant proteins (odorant binding proteins; OBPs) of Manduca sexta. The specificity of each MAb was demonstrated by labeling six immunoblots, each of which contained samples of all six recombinant OBPs. The expression patterns of the six OBPs could be grouped into three classes: (1) one (GOBP1) was expressed in sensilla located throughout each annulus; (2) two (ABPX and ABP2) were expressed in the long sensilla trichoidea bordering a zone that was arranged as an arch on the periphery of each annulus; (3) three (PBP2, PBP3, and GOBP2) were expressed in shorter sensilla occupying a wedge-shaped mid-annular zone of each annulus. In female antennae, sensilla expressing these OBPs were intermixed, and the distinct zonation observed in the male antenna was absent. In males, PBP2 was co-expressed in exactly the same cells of the mid-annular zone as those expressing PBP3 and most of the same cells expressing GOBP2, although its expression overlapped with no or only a few sensilla expressing OBPs of class 1 (GOBP1) or class 2 (ABPX, ABP2). This overlap of expression or lack of overlap between PBP2 and the other OBPs for male antennae was mirrored in female antennae. In view of the restricted spatial expression of OBPs within an annulus and the diversity of possible dimeric combinations of OBPs that arises from the co-expression of multiple OBPs in a given sensillum, OBPs could contribute to the specificity of the olfactory responses of insects. PMID- 12905064 TI - Acute pancreatitis, expression of inducible nitric oxide synthase and defective insulin secretion. AB - A high level of nitric oxide (NO) produced by inducible NO synthase (iNOS) is involved in pancreatic beta-cell dysfunction and apoptosis. In the present study, we examined whether iNOS is also expressed in beta cells after induction of acute pancreatitis (AP) in the rat. Pancreatic islets taken from AP animals and incubated for 60 min in the presence of 20.0 mmol/l glucose showed a decreased insulin secretory response to glucose. The basal insulin release at 1.0 mmol/l glucose was also moderately reduced. Experiments on the dynamics of insulin secretion from perfused pancreas revealed an impairment of both first and second phase of glucose-stimulated insulin release after the induction of AP. Confocal microscopy demonstrated that most of the beta cells in pancreas of rat with AP expressed strong immunoreactivity for iNOS. This was further confirmed by biochemical and Western blot analysis that showed a marked increase in iNOS protein expression and enzyme activity concomitant with a modest reduction in the cNOS protein and activity. Although the mechanisms underlying the defective insulin secretory response of beta cells seen during the early stage of AP are complex, the present finding suggests that the expression of iNOS and a marked iNOS-derived NO production in the beta cells may play at least a contributory role in the impairment of beta-cell function. PMID- 12905065 TI - EphB receptors and ephrinB ligands: regulators of vascular assembly and homeostasis. AB - Eph receptors comprise the largest family of receptor tyrosine kinases consisting of eight EphA receptors (with five corresponding glycosyl-phosphatidyl-inositol anchored ephrinA ligands) and six EphB receptors (with three corresponding transmembrane ephrinB ligands). Originally identified as neuronal pathfinding molecules, genetic loss of function experiments have identified EphB receptors and ephrinB ligands as crucial regulators of vascular assembly, orchestrating arteriovenous differentiation and boundary formation. Despite these clearly defined rate-limiting roles of the EphB/ephrinB system for developmental angiogenesis, the mechanisms of the functions of EphB receptors and ephrinB ligands in the cells of the vascular system are poorly understood. Moreover, little evidence can be found in the recent literature regarding complementary EphB and ephrinB expression patterns that occur in the vascular system and that may bring cells into juxtapositional contact to allow bi-directional signaling between neighboring cells. This review summarizes the current knowledge of the role of EphB receptors and ephrinB ligands during embryonic vascular assembly and discusses recent findings on EphB/ephrinB-mediated cellular functions pointing to the crucial role of the Eph/ephrin system in controlling vascular homeostasis in the adult. PMID- 12905068 TI - Association of the mitochondrial DNA 15497G/A polymorphism with obesity in a middle-aged and elderly Japanese population. AB - Although polymorphism of the mitochondrial DNA 15497guanine/adenine (Mt15497G- >A) leads to the Gly251Ser amino acid replacement on human cytochrome b, it is unknown whether functional alteration of the mitochondrion is induced by the Gly251Ser replacement. To see if an association exists between the Mt15497G-->A polymorphism and obesity, we examined differences in body size, body composition, and regional body fat distribution between the two genotypes in middle-aged and elderly Japanese individuals (825 women and 906 men). The Mt15497 genotype was determined with an automated colorimetric allele-specific DNA probe assay system using the polymerase chain reaction (PCR) method. The Mt15497G-->A polymorphism was detected in 3.5% ( n=60) of all subjects: 2.8% ( n=23) among women and 4.1% ( n=37) among men. After adjusting for age and smoking, we found that body weight, body mass index, waist and hip circumferences, fat mass, fat-free mass, intra abdominal fat and triglycerides were significantly greater in women with the A allele compared with the G allele ( p=0.001-0.025). For men, waist to hip ratio was significantly greater ( p=0.032), and waist circumference, intra-abdominal fat and triglycerides had a trend to be significantly greater ( p=0.062-0.087) in subjects with the A allele compared with the G allele. These data suggest that the Mt15497 polymorphism may be associated with obesity-related variables and lipid metabolism. PMID- 12905069 TI - The HY2 gene as an efficient marker for transposon excision in Arabidopsis. AB - Transposable elements can generate germinal and somatic mutations, and hence represent a powerful tool for the analysis of gene function. Transposons from maize have been adapted to mutagenise the genomes of diverse species. The efficiency of these systems partly relies on the ease with which germinal (i.e. germinally transmitted) or somatic excisions can be detected. Here we describe the use of HY2, a gene that codes for an enzyme involved in the biosynthesis of the phytochrome chromophore, to monitor the excision of a Ds gene-trap element in Arabidopsis thaliana. Taking advantage of the altered germination and de etiolation behaviour of a Ds -tagged hy2 mutant, we have designed an efficient protocol for the recovery of germinal revertants, making HY2 the most precocious excision marker available, to the best of our knowledge. In addition, HY2 is also useful for generating visible sectors in photosynthetic tissues, thanks to the somatic instability of this mutable hy2 allele. PMID- 12905066 TI - Distribution of CGG repeat sizes within the fragile X mental retardation 1 (FMR1) homologue in a non-human primate population. AB - Fragile X syndrome, the most common inherited form of mental retardation, arises in individuals with more than 200 CGG repeats in the 5' untranslated region of the fragile X mental retardation 1 ( FMR1) gene. Although CGG repeat numbers comparable to those found in the normal human population are found in various non human primates, neither the within-species size variation nor the propensity for expansion of the CGG repeat has been described for any non-human primate species. The allele distribution has now been determined for FMR1 (homologue) CGG repeats of 265 unrelated founder females of Macaca mulatta monkeys. Among 530 X chromosomes, at least 26 distinct repeat lengths were identified, ranging from 16 to 54 CGG repeats. Of these alleles 79% have between 25 and 33 CGG repeats. Detailed examination of the CGG region revealed a conserved G (CGG)(2 )G interruption, although in no case was an AGG trinucleotide detected. Two animals carried borderline premutation alleles with 54 CGG repeats, within the region of marginal instability for humans. Thus, M. mulatta may be useful as an animal model for the study of fragile X syndrome. PMID- 12905070 TI - Rapid inactivation of the maize transposable element En/Spm in Medicago truncatula. AB - Transposable elements have been widely used as mutagens in many organisms. Among them, the maize transposable element En/Spm has been shown to transpose efficiently in several plant species including the model plant Arabidopsis, where it has been used for large-scale mutagenesis. To determine whether we could use this transposon as a mutagen in the model legume plant Medicago truncatula, we tested the activity of the autonomous element, as well as two defective elements, in this plant, and in Arabidopsis as a positive control. In agreement with previous reports, we observed efficient excision of the autonomous En/Spm element in A. thaliana. This element was also active in M. truncatula, but the transposition activity was low and was apparently restricted to the tissue culture step necessary for the production of transgenic plants. The activity of one of the defective transposable elements, dSpm, was very low in A. thaliana and even lower in M. truncatula. The use of different sources of transposases suggested that this defect in transposition was associated with the dSpm element itself. Transposition of the other defective element, I6078, was also detected in M. truncatula, but, as observed with the autonomous element, transposition events were very rare and occurred during tissue culture. These results suggest that the En/Spm element is rapidly inactivated in the regenerated plants and their progeny, and therefore is not suitable for routine insertion mutagenesis in M. truncatula. PMID- 12905071 TI - Nucleosome transactions on the Hypocrea jecorina (Trichoderma reesei) cellulase promoter cbh2 associated with cellulase induction. AB - The 5' regulatory region of the cbh2 gene of Hypocrea jecorina contains the cbh2 activating element (CAE) which is essential for induction of cbh2 gene expression by sophorose and cellulose. The CAE consists of two motifs, a CCAAT box on the template strand and a GTAATA box on the coding strand, which cooperate during induction. Northern analyses of cbh2 gene expression has revealed an absolute dependence on induction, but no direct effect of Cre1-mediated carbon catabolite repression. Investigation of the chromatin structure in the wild-type strain showed that, under repressing conditions, there is a nucleosome free region (nfr) around the CAE, which is flanked by strictly positioned nucleosomes. Induction results in a loss of positioning of nucleosomes -1 and -2 downstream of the CAE, thus making the TATA box accessible. Simultaneous mutation of both motifs of the CAE, or of the CCAAT-box alone, also leads to shifting of nucleosome -1, which normally covers the TATA-box under repressing conditions, whereas mutation of the GTAATA element results in a narrowing of the nfr, indicating that the proteins that bind to both motifs in the CAE interact with chromatin, although in different ways. A cellulase-negative mutant strain, which has previously been shown to be altered in protein binding to the CAE, still displayed the induction specific changes in nucleosome structure, indicating that none of the proteins that directly interact with CAE are affected, and that nucleosome rearrangement and induction of cbh2 expression are uncoupled. Interestingly, the carbon catabolite repressor Cre1 is essential for strict nucleosome positioning in the 5' regulatory sequences of cbh2 under all of the conditions tested, and induction can occur in a promoter that lacks positioned nucleosomes. These data suggest that Cre1, the Hap2/3/5 complex and the GTAATA-binding protein are all involved in nucleosome assembly on the cbh2 promoter, and that the latter two respond to inducing conditions by repositioning nucleosome -1. PMID- 12905072 TI - The sterol modifying enzyme LET-767 is essential for growth, reproduction and development in Caenorhabditis elegans. AB - The let-767 gene encodes a protein that is similar to mammalian steroid enzymes that are responsible for the reduction of 17-beta hydroxysteroid hormones. Caenorhabditis elegans is incapable of the de novo synthesis of cholesterol. Therefore, this free-living nematode must extract cholesterol from its environment and modify it to form steroid hormones that are necessary for its survival. C. elegans is unable to survive in the absence of supplemental cholesterol, and is therefore sensitive to cholesterol limitation. We show that a mutation in let-767 results in hypersensitivity to cholesterol limitation, supporting the hypothesis that LET-767 acts on a sterol derivative. Furthermore, let-767 mutants exhibit defects in embryogenesis, female reproduction and molting. Although ecdysone is the major molting hormone in insects, there is as yet no evidence for ecdysone synthesis in C. elegans, suggesting that a different hormone is required for molting in C. elegans. Our results suggest that LET-767 modifies a sterol hormone that is required both for embryogenesis and for later stages of development. PMID- 12905073 TI - Expression and characterization of the type III polyketide synthase 1,3,6,8 tetrahydroxynaphthalene synthase from Streptomyces coelicolor A3(2). AB - Sequence analysis of the metabolically rich 8.7-Mbp genome of the model actinomycete Streptomyces coelicolor A3(2) revealed three genes encoding predicted type III polyketide synthases (PKSs). We report the inactivation, expression, and characterization of the type III PKS homologous SCO1206 gene product as 1,3,6,8-tetrahydroxynaphthalene synthase (THNS). Incubation of recombinant THNS with malonyl-CoA showed THN production, as demonstrated by UV and HPLC analyses. The K(m) value for malonyl-CoA and the k(cat) value for THN synthesis were determined spectrophotometrically to be 3.58+/-0.85 micro M and 0.48+/-0.03 min(-1), respectively. The C-terminal region of S. coelicolor THNS, which is longer than most other bacterial and plant type III PKSs, was shortened by 25 amino acid residues and the resulting mutant was shown to be slightly more active (K(m)=1.97+/-0.19 micro M, k(cat)=0.75+/-0.04 min(-1)) than the wild-type enzyme. PMID- 12905074 TI - Optimization of manganese peroxidase and laccase production in the South American fungus Fomes sclerodermeus (Lev.) Cke. AB - Fomes sclerodermeus produces manganese peroxidase (MnP) and laccase as part of its ligninolytic system. A Doehlert experimental design was applied in order to find the optimum conditions for MnP and laccase production. The factors studied were Cu(2+), Mn(2+) and asparagine. The present model and data analysis allowed us not only to define optimal media for production of both laccase and MnP, but also to show the combined effects between the factors. MnP was strongly influenced by Mn(2+), which acts as an inducer. Under these conditions Cu(2+) negatively affected MnP activity. At 13 days of growth 0.75 U ml(-1) were produced in the optimized culture medium supplemented with 1 mM MnSO(4) and 4 g l(-1) asparagine. The laccase titer under optimized conditions reached maximum values at 16 days of growth: 13.5 U ml(-1) in the presence of 0.2 mM CuSO(4), 0.4 mM MnSO(4) and 6 g l(-1) asparagine. Mn(2+) promoted production of both enzymes. There were important interactions among the nutrients evaluated, the most significant being those between Cu(2+) and asparagine. PMID- 12905075 TI - Cell physiological aspects of the plasma membrane electrogenic H+ pump. AB - This article deals with cell physiological aspects of the plasma membrane electrogenic proton (H+) pump and emphasizes the contribution of the giant algal cells of the Characeae in elucidating the mechanism of the pump. First, a history of the development of intracellular perfusion techniques in characean internodal cells is described, including preparation of tonoplast-free cells. Then, an outline of the hypothesis of the electrogenic H+ pump proposed by Kitasato is introduced, who prophesied the existence of an electric potential generated by an active H+ efflux. Subsequently, a history of finding ATP as the direct energy source of the electrogenic ion pump is presented. Quantitative agreement between the pump current and the ATP-dependent H+ efflux supports the notion that the ion carried by the electrogenic ion pump is H+. The role of the H+ pump in regulation of the cytosolic pH is discussed. Mechanisms of light-induced potential change through photosynthesis-controlled activation of the H+ pump are discussed in terms of changes in the levels of adenine nucleotides and in modulation of the Km value for the ATP of H+-ATPase. Recent progress in the molecular mechanism of the blue-light-induced activation of the H+-ATPase in guard cells is presented. However, there are cases where H+-ATPase activity is inhibited by blue light, indicating the flexibility of the control mechanisms of H+-ATPase activity. Finally, modulation of H+-pumping or H+-ATPase activities in response to environmental factors, such as anoxia, membrane excitation, osmotic and salt stresses, nutrient deficiencies and aluminum toxicity are described. Discussions are presented on the regulation of the electrogenic H+ pump. PMID- 12905076 TI - Regulation of plasma membrane H+-ATPase activity by the membrane environment. AB - The plant plasma membrane H(+)-ATPase is a proton pump which plays a central role in physiological functions such as nutrient uptake and intracellular pH regulation. This pump belongs to the P(3)-type ATPase family and creates an electrochemical gradient across the plasma membrane. The generation of this gradient has a major role in providing the energy for secondary active transport across the plasma membrane. The activity of the proton pump is regulated by the transcriptional and post-translational levels and by membrane environmental factors such as membrane lipids. Several reviews have appeared during the last few years concerning the regulatory mechanism at transcriptional and post translational levels. The plasma membrane H(+)-ATPase requires lipids for activity. This lipid dependency suggests a possible mode of regulation of the H(+)-ATPase via modification of its lipid environment. This review focuses on the regulation of plasma membrane H(+)-ATPase by membrane lipids surrounding H(+) ATPase molecules. PMID- 12905077 TI - Ventriculostomy in a tumor involving the third ventricular floor. AB - Recently, endoscopic management has gotten preference over open surgical treatment in selected cases of intraventricular tumors. Endoscopic third ventriculostomy (ETV) appears unfeasible when tumors extend to the third ventricular floor region due to the risk of perforators and injury to the basilar artery. We report the case of a 12-year-old male with symptoms of acute, chronic, raised intracranial pressure. Computed tomography (CT) and magnetic resonance imaging (MRI) showed a posterior third ventricular tumor involving the aqueductal and floor regions. The ETV was done after clearing the floor by partial tumor resection, keeping the dorsum sellae as the major anatomical landmark. The patient improved satisfactorily and was given adjuvant radiotherapy, and the need for an external shunt was completely eliminated. We conclude that ETV appears worth trying, even in third ventricular tumors involving the floor region if they can be cleared from the tumor keeping the dorsum sellae as the major anatomical landmark. PMID- 12905079 TI - Cognitive development in object manipulation by infant chimpanzees. AB - This study focuses on the development of spontaneous object manipulation in three infant chimpanzees during their first 2 years of life. The three infants were raised by their biological mothers who lived among a group of chimpanzees. A human tester conducted a series of cognitive tests in a triadic situation where mothers collaborated with the researcher during the testing of the infants. Four tasks were presented, taken from normative studies of cognitive development of Japanese infants: inserting objects into corresponding holes in a box, seriating nesting cups, inserting variously shaped objects into corresponding holes in a template, and stacking up wooden blocks. The mothers had already acquired skills to perform these manipulation tasks. The infants were free to observe the mothers' manipulative behavior from immediately after birth. We focused on object object combinations that were made spontaneously by the infant chimpanzees, without providing food reinforcement for any specific behavior that the infants performed. The three main findings can be summarized as follows. First, there was precocious appearance of object-object combination in infant chimpanzees: the age of onset (8-11 months) was comparable to that in humans (around 10 months old). Second, object-object combinations in chimpanzees remained at a low frequency between 11 and 16 months, then increased dramatically at the age of approximately 1.5 years. At the same time, the accuracy of these object-object combinations also increased. Third, chimpanzee infants showed inserting behavior frequently and from an early age but they did not exhibit stacking behavior during their first 2 years of life, in clear contrast to human data. PMID- 12905080 TI - A Capuchin monkey (Cebus apella) recognizes when people do and do not know the location of food. AB - In a previous study, Kuroshima and colleagues demonstrated that capuchin monkeys (Cebus apella) learned to discriminate between a "knower" who inspected a box for food, and a "guesser" who did not. The aim of the present study was to specify whether the subjects learned a simple conditional discrimination or a causal relationship that seeing leads to knowing. In experiment 1, we introduced five types of novel containers to two subjects. Each container was of different shape and color. The subjects gradually learned to reach toward the container the knower suggested. In experiment 2, we diversified the behavior of the knower and the guesser. In experiment 3, in order to eliminate the possibility of discrimination based on differences in the magnitude and the complexity of two trainers, we equated their behaviors. One subject adapted to the novel behaviors of the knower and the guesser, successfully discriminating the two trainers. Thus this monkey clearly learned to use the inspecting action of the knower and the non-inspecting action of the guesser as a discriminative cue to recognize the baited container. This result suggests that one capuchin monkey learned to recognize the relationship between seeing and knowing. PMID- 12905078 TI - A prospective clinical comparison of rectangular titanium cages and iliac crest autografts in anterior cervical discectomy and fusion. AB - The complications of autogenous bone grafting cause spinal surgeons to seek alternative methods for cervical spinal fusion. This prospective study was conducted to evaluate the safety and efficacy of rectangular titanium cages as compared to the widely performed iliac crest autograft fusion. Thirty-six patients with cervical disc disease in whom an anterior cervical approach was indicated for discectomy were included in a prospective controlled study protocol with 1-year follow-up. The first 18 consecutive patients received iliac crest autograft, while the next 18 received rectangular titanium cages. According to Odom's criteria, 15 of 18 (83%) patients in both groups experienced good to excellent functional recovery. According to the patient satisfaction index, 17 of 18 (94%) in both groups were satisfied. There were no significant differences in neck or arm pain. Fusion was present after 1 year in 16 of 18 (89%) patients in the iliac crest autograft group and 15 of 18 (83%) in the rectangular titanium cage group. In the autograft group, one case of pseudarthrosis was present, and marked hip pain was observed in four patients. There were no implant-related complications in the cage group. The authors conclude that titanium cages in anterior cervical discectomy constitute a safe and efficient alternative to iliac crest bone autograft. PMID- 12905081 TI - Genetic deficiency of a mitochondrial aldehyde dehydrogenase increases serum lipid peroxides in community-dwelling females. AB - Mitochondrial aldehyde dehydrogenase 2 (ALDH2) plays a major role in acetaldehyde detoxification. The alcohol sensitivity is associated with a genetic deficiency of ALDH2. We and others have previously reported that such a deficiency influences the risk for late-onset Alzheimer's disease (LOAD), hypertension, and myocardial infarction. Then we tried to find phenotypes to which the ALDH2 polymorphism contributes by conducting several evaluations including biochemical and functional analyses of various tissues in a community-dwelling population. Several serum proteins, lipids, and lipid peroxides (LPO) levels showed differences between the nondefective (ALDH2*1/1) and defective (ALDH2*1/2 and ALDH2*2/2) ALDH2 individuals. However, alcohol-drinking behavior is known to affect these evaluations. Thus, we excluded the effects of alcohol-drinking behavior from the association with the ALDH2-deficient genotype through correction and found that the concentration of LPO was significantly lower in the nondefective ALDH2 females than the defective females. The effect of frequent alcohol-drinking behavior in males seems to override the phenotype of the high serum LPO level. These results indicate that the ALDH2 deficiency may enhance oxidative stress in vivo. Thus, these findings suggest that ALDH2 functions as a protector against oxidative stress and the decrease in protection may influence the onset of AD, hypertension, and myocardial infarction. PMID- 12905082 TI - A novel locus for parietal foramina maps to chromosome 4q21-q23. AB - Parietal foramina [PFM], inherited usually in an autosomal dominant mode, is an extremely rare developmental defect characterized by a symmetrical, oval hole in the parietal bone. It can be present as either an isolated or a syndromic feature. PFM types 1 and 2 (PFM1 and PFM2) have been found to be caused by mutations in the MSX2 and ALX4 genes, located to chromosomes 5 and 11, respectively. After exclusion of both the above loci in a large Chinese pedigree with autosomal dominant PFM, a genome-wide search revealed a linkage of the PFM to markers at the 4q21-q23 region. The maximum LOD score from two-point linkage analysis is 3.87 for marker D4S2961. Analysis of co-segregated haplotype localized the region to a 20-cM interval that flanks D4S392 and D4S2945. Therefore, we concluded that the PFM in the family is a new PFM locus. Although three genes, BMPR1B, PP1 and IBSP, are located to 4q21-q25 and their functions are related to bone morphogenesis, no mutations were identified by sequencing analysis of their exons. PMID- 12905085 TI - The effects of massage on intra muscular temperature in the vastus lateralis in humans. AB - The aim of the current investigation was to evaluate the effect of different durations of massage, and ultrasound treatment, on the temperature of the vastus lateralis muscle in males. Deep effleurage massage of the vastus lateralis was performed on seven healthy males (mean +/- SD; age; 28 +/- 4 years, height 1.84 +/- 0.05 m, body mass 85.7 +/- 12 kg) for 5, 10 and 15 min periods. A 5-min period of ultrasound at 45 KHz was also completed by all subjects. Intra muscular temperature (at 1.5, 2.5 and 3.5 cm) and thigh skin temperature were assessed pre and post treatment. Heart rate was monitored continuously throughout all conditions. Pre treatment intra muscular temperature increased as depth of measurement increased (p = 0.00002). Changes in muscle temperature at 1.5 and 2.5 cm were significantly greater following massage than ultrasound (p < 0.002). No significant differences between massage treatments and ultrasound were noted when intra muscular temperature was measured at 3.5 cm (p > 0.05). Massage also significantly increased both heart rate and thigh skin temperature compared to ultrasound (p < 0.005). Increases in intra muscular temperature, heart rate and thigh skin temperature were the same irrespective of massage duration. These data suggest that massage and ultrasound have only limited effects on deep muscle temperature. As a result such approaches may not be suitable as a preparation strategy for exercise. PMID- 12905086 TI - Perfusion distribution between and within muscles during intermittent static exercise in endurance-trained and untrained men. AB - We have recently shown that muscle perfusion varies between different quadriceps femoris muscles during submaximal exercise in humans. In animals, endurance training changes perfusion distribution between muscles during exercise. Whether the same is observed in humans is currently unknown. Therefore, we compared perfusion levels between different parts of the quadriceps femoris muscle group during one-legged intermittent static exercise in seven endurance-trained and seven untrained men. Muscle perfusion was measured using positron emission tomography with [ 15O]-H 2 O. In addition, relative dispersion of perfusion (standard deviation within a region/mean within a region x 100 %) within each muscle region was calculated as an index of perfusion heterogeneity within the muscles. Muscle perfusion tended to be lower in endurance-trained men (p = 0.16) and it was also different between the regions (p < 0.001). However, perfusion distributed similarly between the groups (p = 0.51). Relative dispersion of perfusion within the muscles was lower in endurance-trained men (p = 0.01) and it was also different between muscles (p < 0.001). These results suggest that endurance training does not alter perfusion distribution between muscles, but it decreases perfusion heterogeneity within the muscles. PMID- 12905087 TI - The effects of exercise training on markers of endothelial function in young healthy men. AB - This study investigated the effects of fitness and of acute exercise on a range of markers of endothelial function in young, healthy adult male subjects who were classified on the basis of maximum oxygen consumptions as being fit VO(2 peak) 71 +/- 2 [ml x min (-1)] x kg (-1) or sedentary VO(2 peak) 53 +/- 2 [ml x min (-1)] x kg (-1). Fit and sedentary subjects had similar resting plasma levels of von Willebrand factor (vWF) and thrombomodulin (TM). Acute maximal aerobic exercise doubled plasma vWF in fit subjects but had no effect in the sedentary population; plasma TM rose with acute exercise in each group but to a greater extent in the fit population. Fit subjects also had higher numbers of circulating endothelial cells (CECs) at rest and exhibited substantially greater forearm reactive hyperaemia responses following a standardized period of arterial occlusion. A cohort of sedentary subjects was given a 5-week training programme of moderate aerobic exercise on a cycle ergometer. Following this, absolute fitness was increased by only 8 % but reactive hyperaemia responses rose to values similar to those in the chronically fit group. The results suggest that both acute and chronic exercise increase endothelial turnover. Chronic exercise is also associated with enhanced endothelium-dependent dilator function and this effect becomes maximal after only a short period of moderate training. PMID- 12905088 TI - Acute hormonal and neuromuscular responses and recovery to forced vs maximum repetitions multiple resistance exercises. AB - Acute hormonal and neuromuscular responses and recovery three days after the exercises were examined during the maximum repetitions (MR) and forced repetitions (FR) resistance exercise protocols in 16 male athletes. MR included 4 sets of leg presses, 2 sets of squats and 2 sets of knee extensions (with 12 RM) with a 2-min recovery between the sets and 4 min between the exercises. In FR the initial load was chosen to be higher than in MR so that the subject could not lift 12 repetitions per set by himself. After each set to failure the subject was assisted to perform the remaining repetitions to complete the 12 repetitions per set. Thus the exercise intensity was greater in FR than in MR. Both loading protocols led to the great acute increases (p < 0.05 - 0.001) in serum testosterone, free testosterone, cortisol and GH concentrations. However, the responses in cortisol (p < 0.05) and GH (p < 0.01) were larger in FR than in MR. The decrease of 56.5 % (p < 0.001) in maximal isometric force in FR was greater (p < 0.001) than that of 38.3 % in MR (p < 0.001) and force remained lower (p < 0.01) during the recovery in FR compared to MR. The larger decrease in isometric strength in FR than in MR was also associated with the decreased maximal voluntary EMG of the loaded muscles. The data indicate that the forced repetition exercise system induced greater acute hormonal and neuromuscular responses than a traditional maximum repetition exercise system and therefore it may be used to manipulate acute resistance exercise variables in athletes. PMID- 12905089 TI - Resistance training improves glycaemic control in obese type 2 diabetic men. AB - The purpose of this investigation was to determine whether moderate intensity resistance training (RT) improves glycaemic control in obese, type 2 diabetic men. Eighteen subjects were randomly assigned to a 10-week RT program, or a non training control group (C). Glycosylated haemoglobin (HbA 1c ), fasting glucose and insulin, glucose and insulin 120 minutes (2h) after a 75 g oral glucose load, body composition and muscular strength and endurance were measured before and after the 10-week experimental period. In the RT group fasting glucose and insulin decreased with training (p < 0.05) and decreases in HbA 1c approached significance (p = 0.057). 2-h glucose and insulin did not change in either group. Fat free mass (FFM) increased by 3.5 % after RT but was unchanged in the controls. Fat mass (FM) increased 6.9 % in C but was unchanged in RT. Percent body fat was unchanged in both groups. Muscular strength and endurance increased by 25 to 52 % in the RT group but was unchanged in controls. Changes in fasting glucose and HbA 1c were inversely related to changes in FFM. These results suggest that RT is an effective form of exercise training which modestly improves glycaemic control and lowers fasting insulin levels in obese type 2 diabetics. PMID- 12905090 TI - Finger flexors fatigue in trained rock climbers and untrained sedentary subjects. AB - The present series of experiments were conducted to access the surface EMG frequency parameters during repeated fingertip isometric contractions to determine if they can be used as a fatigue index under specific grip used in rock climbing. Electromyograms of the finger flexors and extensors were characterised in ten elite climbers and ten non-climbers. The exercise consisted in reaching 80 % of maximal isometric finger force as quickly as possible intermittently with a 5-s contraction followed by 5-s of rest until exhaustion (i. e. when the subject was unable to maintain 80 - 70 % MVC force range for the 5 s). The results clearly indicate that expert climbers performed significantly greater fingertip force than sedentary subjects (420 +/- 46 N vs. 342 +/- 56 N). This force was maintained during twelve repetitions (12.88 +/- 4.96) in sedentary subjects, whereas the climbers maintained the force during nineteen repetitions (19.33 +/- 4.84). The median frequency of both the flexor and extensor EMG power spectra decreased during fatiguing isometric contractions, but at different rates in climbers and non-climbers. In non-climbers, the results replicated previous findings, whereas in climbers the results were novel. PMID- 12905091 TI - Sport specific performance diagnosis in rowing: an incremental graded exercise test in coxless pairs. AB - The aim of the present study was to develop and validate an incremental graded exercise test performed in the rowing boat(coxless pair) in order to give specific performance evaluation data. Furthermore, an attempt was made to transfer these data to training recommendations. Thirty-four female rowers of national and international level performed a 4 x 6 min incremental graded exercise test GXT(boat) in coxless pairs on a lake (6 km, no wind, no waves). The boat velocity on the water (V; measured by a speedometer PACE COACH) was increased continuously from 3.55 m x s-I to 4.03 m x s-1. The individual anaerobic threshold(IAT) was determined by means of the lactate (LA) kinetics during and after exercise. Within 28 days all subjects performed arowing ergometry test GXT(ergo); Gjessing rowing ergometer: 40 watts increments every 3 min) as well as 70 min of constant endurance training in the boat in moderate velocity (ET; n- 10 pairs because of changing weather conditions). Results for V/LAIHR at IAT are: GXT(boat): 3.84+/- 0.10 m x s-1/2.44 +/- 0.66 mmol x 1-1/172 + 11 min(-1); GXT(ergo): 206+/-10 watts/2.53 +0.40 mmol x 1-171 +/- 10 min-' (means+/-SD). The Spearman rank order test showed significant correlations for HR (p < 0.001) and the mean performances of the coxless pairs (p < 0.05). A'5 % lower V during ET lead to a 10% lower HR and a 30% lower LA compared to the values at IAT. In conclusion, both a performance specific evaluation and velocity oriented control of training are possible by means of a 4 x 6 min incremental graded exercise test in coxlesspairs. However, this test on the water requires almost perfect weather conditions. The HR recommendations based on GXT (ergo) were confirmed during GXT(boat). PMID- 12905092 TI - Biological variation in variables associated with exercise training. AB - To be able to identify a training induced change in a certain variable, it is necessary to know the background variation. In this study the coefficient of variation (total, between-subjects, within-subjects), the relative sources of variance (between-subjects and within-subjects), and the critical difference (within-subjects) were estimated in four categories of variables (performance and physiological variables, metabolic and hormonal variables, immunological variables, and mood state variables) in 15 moderately trained male runners measured on three different occasions over a period of 7 weeks. In the performance and physiological variables, 78.9 % of the variance was due to variation between subjects and they had the lowest critical difference (11.9 %). In contrast, the metabolic and hormonal variables had the highest critical difference (59.9 %) and 53.4 % of the variance was due to variations within subjects. The immunological and psychological variables had about two thirds of the variance arising from variation between subjects. However, the critical difference for the immunological variables was high (47.4 %), while it was relatively low for the psychological variables (26.8 %). The low critical difference and variation within subjects of the psychological and in particular the performance and physiological variables indicate that they may be beneficial as primary markers of training induced changes. PMID- 12905093 TI - Reduction of blood pressure response during strength training through intermittent muscle relaxations. AB - The increase in blood pressure during training is a disadvantage of strength training in the elderly. To reduce this effect it is generally recommended to apply lower levels of relative muscle strength with longer contraction durations or higher number of repetitions (continuous mode (CM), e. g. 50% of maximal strength, 10 to 15 repetitions without pauses). Alternatively, higher contraction forces could be combined with frequent periods of muscle relaxation and fewer repetitions (intermittent mode (IM), e. g. 80% of maximal strength, 8 repetitions consisting of 1.5 s concentric contraction, 1.5 s eccentric contraction and 3 s pause). We compared the blood pressure effects of both approaches during leg press exercise in two age groups (10 subjects aged 22 to 42 y, 9 subjects aged 60 to 72 y). Blood pressure was measured continuously by a non-invasive method (FINAPRES, Ohmeda 2300, Englewood USA). RESULTS: 1. The age of the subjects had no significant influence on the slopes of blood pressure increase during the different exercise modes. 2. The frequent insertion of short (3 s) periods of muscle relaxation (IM) decreased the blood pressure response more effectively than a reduction in contraction strength alone (CM). Short muscle relaxations have an immediate, mechanical effect on blood pressure and they allow a metabolic recovery which attenuates the trend of blood pressure increase. PMID- 12905094 TI - Blood lactate concentration/heart rate relationship: laboratory running test vs field roller skiing test. AB - Laboratory treadmill running tests are commonly used to assess the effects of training programs and to prescribe training intensity for cross-country skiers. The present study compared the blood lactate concentration ([La])/heart rate (HR) relationship obtained during a laboratory treadmill running test and used to prescribe training intensities, to the same relationship obtained during a field roller skiing test in ten young male cross-country skiers of national level. The laboratory test consisted of a maximal incremental protocol involving 3 min steps during which metabolic and cardio-respiratory parameters were measured. The field test comprised four loops (4 km long) at different intensities from warm-up speed to maximum aerobic speed during which HR and [La] were measured. For a given HR, [La] values obtained during the field test were significantly higher compared with those measured in the laboratory test (mean difference from 0.2 mmol x l -1 for HR = 147 bpm, to 4.9 mmol x l -1 for HR = 182 bpm; p < 0.05). Exercise specificity, i. e. upper body involvement for roller skiing, as well as protocol characteristics, particularly longer step duration, could explain this discrepancy. The results of the present study highlight the importance of exercise and protocol specificity for [La]/HR relationship currently used to prescribe exercise intensity in cross-country skiing. PMID- 12905095 TI - Effect of carbohydrate feeding during recovery from prolonged running on muscle glycogen metabolism during subsequent exercise. AB - This study examined the effect of carbohydrate (CHO) intake during a 4 h recovery from prolonged running on muscle glycogen metabolism during subsequent exercise. On 2 occasions, 7 male subjects ran for 90 min at 70 % maximum oxygen uptake VO(2 max) on a motorized treadmill (R1) followed by a 4 h rest period (REC) and a 15 min run (R2) consisting of 5 min at 60 % and 10 min at 70 % VO(2 max) During REC, each subject ingested a total of 2.7 l of an isotonic solution containing either 50 g of CHO (LOW) or 175 g of CHO (HIGH). Biopsy samples were obtained from the vastus lateralis immediately after R1, REC and R2. During REC, a higher muscle glycogen resynthesis was observed in HIGH when compared with LOW trial (75 +/- 20 vs. 31 +/- 11 mmol x kg dry matter (dm) -1, respectively; p < 0.01). Muscle glycogen utilization during R2 was similar between the HIGH and LOW trials (39 +/ 10 vs. 46 +/- 11 mmol x kg dm -1, respectively). These results suggest that ingestion of a large amount of CHO at frequent intervals during recovery from exercise does not affect the rate of muscle glycogen utilization during subsequent exercise. PMID- 12905096 TI - Adherence and attrition with intermittent and continuous exercise in overweight women. AB - Few people are active to the levels recommended by Healthy People 2010. Intermittent exercise has been promoted as an exercise prescription which may enable more people to meet recommended guidelines. However, few data are available on intermittent exercise over the long-term. The purpose of the current study was to compare the effects of long-term (72 weeks) continuous (CON) and intermittent (INT) exercise on attrition and adherence in previously sedentary, moderately obese females. Participants were randomized to continuous walking at 60 to 75% of maximum aerobic capacity, 3 days per week, 30 minutes per session, or intermittent exercise for two 15-minute sessions, 5 days per week. Adherence was calculated as the number of sessions completed compared to the number of sessions prescribed. At 12-week intervals, attrition was calculated as the number of participants in the study compared to the total number of participants originally enrolled. For the participants who completed the study, body weight decreased for CON from 80.17 +/- 5.75 kg at baseline to 79.70 +/- 5.40 at 16 months (p < 0.05). For INT, body weight did not change from baseline (85.85 +/- 13.13 kg) to 16 months (85.05 +/- 12.90 kg). By design, INT walked significantly (p < 0.05) further (819 +/- 128 km) compared to CON (527 +/- 46 km). Attrition was 58% for both groups baseline to 72 weeks. However, attrition was greater for CON (38 %) compared to INT (16%) in the first 24 weeks. Adherence was excellent for both groups (> 83%) throughout the study. These results suggest that intermittent and continuous exercise both have considerable attrition rates within 72 weeks of exercise initiation; however, the pattern of attrition differs considerably. That is, it appears that intermittent exercise may reduce attrition in the first 24 weeks of an exercise program; however, attrition does not appear to be different than continuous exercise at 72 weeks. PMID- 12905097 TI - Analysis of 3D kinematics concerning three different clubs in golf swing. AB - Although many professionals have produced books or videotapes which offer a novel approach to the game, a review of the scientific literature reports limited research evaluating the actual biomechanics of the golf swing in comparison with other sports. The aim of this study was to investigate the influence of kinematic pattern in golf swing using three different clubs: driver, five-iron, pitching wedge. These three golf clubs have been chosen for their available range of ball flight. The measures of kinematic data during swing were established with the optoelectronic system VICON (Oxford's Metrics, Oxford, UK) with five cameras operating at 50 frames per second. Clubhead speed was measured using a swing made detector (Bell-Tronics, Ltd, Covington, USA). Seven right-handed male golfers with a high level of skill participated in the study. The results showed that there was an identical timing (movement time and proportion for each phase of the swing) between the three clubs tested, but the kinematics and the clubhead speed were different depending on the three different clubs used. PMID- 12905098 TI - Cognitive performance, SPECT, and blood viscosity in elderly non-demented people using Ginkgo biloba. AB - The aging process is associated with several cognitive alterations. This study looks at the effects of taking dried extract of Ginkgo biloba, which has been used in several countries in an attempt to minimize these effects. The subjects were 48 men aged 60 - 70 matched between control and experimental groups for educational level. Evaluation was based on a number of neuropsychological tests in an attempt to cover the largest possible number of functions including Single Photon Emission Computer Tomography (SPECT) and measures of blood viscosity. The study was run on a double-blind basis with placebo and Ginkgo biloba groups evaluated over a period of 8 months. After treatment, the experimental group showed a reduction in blood viscosity, improved cerebral perfusion in specific areas and improved global cognitive functioning. The control group showed the opposite - higher blood viscosity, a reduction in cerebral perfusion (in specific areas), and cognitive deterioration in different functions. Although the mechanisms by which Ginkgo biloba may contribute to overall enhancement of the parameters evaluated have not been specified, this plant extract certainly appears to be effective in the treatment of cognitive deficits in older people. Further research into its use is called for on the basis of the results obtained here. PMID- 12905099 TI - Predictors of therapeutic effects in amitriptyline treatment--1. Plasma drug levels. AB - OBJECTIVE: To analyze the spectrum of relationships between clinical effects of amitriptyline (At) treatment after 2 and 4 weeks (wks) and plasma levels of At, nortriptyline (Nt), At+Nt, demethylation rate of At, treatment modalities, age, and gender. METHODS: Patients with major depression (ICD 10: F31-F33) and a HAMD 21 total score of 15-41 received At on a dosage schedule chosen by the doctor for at least 4 wks. Plasma drug levels were assessed at baseline and at wks 2 and 4. RESULTS: Of the 58 patients enrolled in the study, 47 (15males, 32 females) were eligible for statistical analysis. An early response by wk 2 (decrease in HAMD-21 score of at least 50 % from baseline) was observed in 34.0 % of patients, and after 4 wks, the response rate was 63.8 % (males 86.6 %, females 53.1 %). There was a low, negative, and significant correlation between percent reduction in HAMD and steady state At concentration only at wk 2 (n = 47 r Sp. = -0.306 p < 0.05). However, the correlation was dependent on the degree of At demethylation and treatment modalities. A ratio of Nt/At >1 was observed in 23 patients; of these, 11 (47.8 %) were non-responders by wk 4. A low rate of demethylation (Nt/At 1 group, the non responders were distributed in the whole range of observed At plasma concentrations (20-150 ng/ml). Only in patients with Nt/At 95% O2) conditions, exercise-induced Tb depression was abolished. Administration of CPT similarly abolished decreased Tb due to both exercise and hypoxia. Trials using Dipsosaurus dorsalis indicate that elevated ambient oxygen during exercise does not influence blood pH or lactate accumulation, suggesting that these factors do not initiate changes in thermoregulatory setpoint following exhaustive exercise. We suggest that when oxygen is limiting, a decrease in arterial oxygen may trigger the release of adenosine, thereby altering the thermoregulatory setpoint. PMID- 12905121 TI - Body temperature null distributions in reptiles with nonzero heat capacity: seasonal thermoregulation in the American alligator (Alligator mississippiensis). AB - Regulation of body temperature may increase fitness of animals by ensuring that biochemical and physiological processes proceed at an optimal rate. The validity of current methods of testing whether or not thermoregulation in reptiles occurs is often limited to very small species that have near zero heat capacity. The aim of this study was to develop a method that allows estimation of body temperature null distributions of large reptiles and to investigate seasonal thermoregulation in the American alligator (Alligator mississippiensis). Continuous body temperature records of wild alligators were obtained from implanted dataloggers in winter (n=7, mass range: 1.6-53.6 kg) and summer (n=7, mass range: 1.9-54.5 kg). Body temperature null distributions were calculated by randomising behavioural postures, thereby randomly altering relative animal surface areas exposed to different avenues of heat transfer. Core body temperatures were predicted by calculations of transient heat transfer by conduction and blood flow. Alligator body temperatures follow regular oscillations during the day. Occasionally, body temperature steadied during the day to fall within a relatively narrow range. Rather than indicating shuttling thermoregulation, however, this pattern could be predicted from random movements. Average daily body temperature increases with body mass in winter but not in summer. Daily amplitudes of body temperature decrease with increasing body mass in summer but not in winter. These patterns result from differential exposure to heat transfer mechanisms at different seasons. In summer, alligators are significantly cooler than predictions for a randomly moving animal, and the reverse is the case in winter. Theoretical predictions show, however, that alligators can be warmer in winter if they maximised their sun exposure. We concluded that alligators may not rely exclusively on regulation of body temperature but that they may also acclimatise biochemically to seasonally changing environmental conditions. PMID- 12905122 TI - Foot-mediated incubation: Nazca booby (Sula granti) feet as surrogate brood patches. AB - Incubation in most avian species involves transferring heat from parent to egg through a highly vascularized brood patch. Some birds, however, do not develop a brood patch. Unusual among birds, these species hold their eggs under the webs of their feet, but the role of the feet in heat transfer is uncertain. Often the webs are positioned between the feathered abdomen and the egg during incubation, suggesting that either the abdomen, the feet, or both could transfer heat to the egg. We studied heat transfer from foot webs to eggs during incubation in Nazca boobies by spatially separating the feet from the abdomen using an oversized egg. We found that feet transfer heat to eggs independently of any heat that may be transferred from the abdomen. In addition, we found that incubating boobies had significantly greater vascularization in their foot webs, measured as a percentage of web area covered by vessels, than nonincubating boobies. We also found that males, whether incubating or nonincubating, had significantly less web vascularization than females. We concluded that vascularized Nazca booby feet function in the same way during incubation that vascularized brood patches do, acting as surrogate brood patches. PMID- 12905123 TI - Carotenoids in eggs and plasma of red-legged partridges: effects of diet and reproductive output. AB - Carotenoids are important dietary constituents in birds. They serve as pigments and play numerous physiological roles in both the laying hen and developing embryo. However, factors determining the absorption of carotenoids and their allocation to different functions are numerous and complex, and causal relationships are generally poorly known. Our objective was to determine the degree to which carotenoid levels in egg yolks and the plasma of hens were influenced by differences in diet and reproductive output in captive red-legged partridges. Carotenoid concentrations were measured by high performance liquid chromatography in two feeds (high and low carotenoid content) and in yolks and plasma of hens near the start and end of laying. Early in the laying season, plasma and yolk carotenoids varied with diet and were correlated with one another. Late in the season, a dietary effect was evident only for yolks, and there was no relationship between plasma and egg levels of individual hens. However, plasma carotenoids at the end of laying were strongly correlated with the number of eggs that had been laid. Dietary availability, although important, could explain some variation in carotenoid levels in plasma and egg yolks only in the context of reproductive history. PMID- 12905124 TI - Variation in energy expenditure among black-legged kittiwakes: effects of activity-specific metabolic rates and activity budgets. AB - We sought to determine the effect of variation in time-activity budgets (TABs) and foraging behavior on energy expenditure rates of parent black-legged kittiwakes (Rissa tridactyla). We quantified TABs using direct observations of radio-tagged adults and simultaneously measured field metabolic rates (FMR) of these same individuals (n=20) using the doubly labeled water technique. Estimated metabolic rates of kittiwakes attending their brood at the nest or loafing near the colony were similar (ca. 1.3 x basal metabolic rate [BMR]), although loafing during foraging trips was more costly (2.9 x BMR). Metabolic rates during commuting flight (7.3 x BMR) and prey-searching flight (6.2 x BMR) were similar, while metabolic rates during plunge diving were much higher (ca. 47 x BMR). The proportion of the measurement interval spent foraging had a positive effect on FMR (R2=0.68), while the combined proportion of time engaged in nest attendance and loafing near the colony had a negative effect on FMR (R2=0.72). Thus, more than two-thirds of the variation in kittiwake FMR could be explained by the allocation of time among various activities. The high energetic cost of plunge diving relative to straight flight and searching flight suggests that kittiwakes can optimize their foraging strategy under conditions of low food availability by commuting long distances to feed in areas where gross foraging efficiency is high. PMID- 12905125 TI - Influence of normal daytime fat deposition on laboratory measurements of torpor use in territorial versus nonterritorial hummingbirds. AB - Fat deposition and torpor use in hummingbirds exhibiting distinct foraging styles should vary. We predicted that dominant territorial hummingbirds will use torpor less than subordinate nonterritorial species because unrestricted access to energy by territory owners allows for fat storage. Entry into torpor was monitored using open-flow respirometry on hummingbirds allowed to accumulate fat normally during the day. Fat accumulation was measured by solvent fat extraction. Territorial blue-throated hummingbirds (Lampornis clemenciae) had the highest fat accumulation and used torpor only 17% of the time. Fat storage by L. clemenciae averaged 26% of lean dry mass (LDM) in 1995 and 18% in 1996, similar to that measured for other nonmigratory birds. Fat storage by magnificent hummingbirds (Eugenes fulgens; trapliner) and black-chinned hummingbirds (Archilochus alexandri; nectar robber) averaged 19% and 16% of LDM, respectively, and they used torpor frequently (64% and 92% of the time, respectively). All species initiated torpor if total body fat dropped below 10% of LDM, indicating the existence of a torpor threshold. The ability of L. clemenciae to store enough fat to support nighttime metabolism is likely an important benefit of territoriality. Likewise, frequent torpor use by subordinates suggests that natural restrictions to energy intake can impact their energy budget, necessitating energy conservation by use of torpor. PMID- 12905126 TI - Differential effects of multiple short day lengths on body weights of gonadectomized siberian hamsters. AB - Siberian hamsters (Phodopus sungorus) maintained under simulated natural photoperiods exhibit marked reductions in body weight as day lengths decrease in summer and fall. This experiment assessed whether the component of the seasonal body weight rhythm that is independent of gonadal hormones exhibits a graded dependence on decreasing day lengths or whether the entire program of weight loss is triggered by the crossing of a single critical day length in late summer. Male hamsters born into a photoperiod with 13 h light and 11 h dark (i.e., 13L : 11D) were castrated and transferred to simulated natural photoperiod for early April at 40 degrees N latitude. At the summer solstice (15L : 9D) some hamsters remained on that photoperiod whereas others experienced gradual decreases in day length. Three additional groups were moved to static photoperiods when day lengths had subsequently declined to 13L : 11D, 11L : 13D, or 9L : 15D, respectively. Day lengths decreasing to 13L : 11D were sufficient to suppress body weight but were less inhibitory than further decreases in day length. Hamsters identified as photononresponsive on the basis of daily activity rhythms increased body weight monotonically. These results establish that steroid independent modulation of body weight depends on photoperiod in a graded fashion. PMID- 12905127 TI - Seasonal changes in the physiology of male Virginia opossums (Didelphis virginiana): signs of the Dasyurid semelparity syndrome? AB - Semelparity, which is multiplying once in a lifetime, is a rare reproductive strategy among mammals. Several species of the marsupial family Dasyuridae experience 100% male mortality following an intense mating period. We investigated seasonal physiological changes that may be associated with early mortality in the male Virginia opossum (Didelphis virginiana; Didelphidae) and compared these changes with those of semelparous, male dasyurids. Free-ranging male Virginia opossums (n=36) were collected during 2001 at the Oklahoma State University Cross Timbers Experimental Range. Seasonal data were collected on hematological, morphological, and helminth parameters of these individuals. We used one-way ANOVA to determine whether there were seasonal differences among means for each parameter. It appeared that male Virginia opossums experienced some physiological changes similar to those of male dasyurids exhibiting semelparity. All males collected in summer (August) were juveniles of the year. Lack of adult males in August suggests high mortality of this cohort during the breeding season. Opossum characteristics exhibiting the dasyurid semelparity syndrome included packed cell volume, adrenal mass, and helminth numbers. Minor lymphocytopenia, neutrophilia, and testosterone concentrations also were similar to semelparous dasyurids. However, a lack of change in serum cortisol concentration and body mass and dynamics in immunoglobulin protein, serum protein, and testes mass were not consistent with previous reports of semelparous dasyurid physiology. Evolutionary divergence and differences in breeding behavior between dasyurids and didelphids may be responsible for the lack of consistency between the taxa. PMID- 12905128 TI - Weaning mass affects changes in body composition and food intake in harbour seal pups during the first month of independence. AB - In phocid seals, the transition to nutritional independence is abrupt, with females abandoning their offspring after weaning and returning to sea. We hypothesized that body size at weaning may play an important role in the nature of this transition. We studied the changes in body composition and water flux of newly weaning harbour seals over the first 4-6 wk postweaning. Thirty-three pups were dosed with deuterium oxide to estimate total body water (TBW) and a subset of 24 was dosed twice to estimate changes in body composition and water flux. All pups lost body mass over the study period, but TBW increased during the period of mass loss, indicating continued lean tissue growth. Combined data from this and our early study indicated that heavy (>median mass) pups were relatively fatter (41.0% vs. 37.1%) and had significantly greater total body energy at weaning than did light (< or = median mass) pups. Percentage TBW declined linearly over time in light pups but was constant in heavy pups for the first 19 d postweaning and then declined linearly. Both the temporal pattern and composition of mass loss differed between light and heavy pups. Estimated food intake increased in the second 2 wk of study compared to the first 2 wk, in both light and heavy pups, reflecting increased foraging success but at levels still insufficient to meet daily expenditures of most individuals. PMID- 12905129 TI - Difference in time to detection: a simple method to differentiate catheter related from non-catheter-related bloodstream infection in immunocompromised pediatric patients. AB - Current methods for diagnosis of catheter-related infection (CRI) are cumbersome and may require removal of the central venous catheter (CVC). A prospective study was conducted to validate the difference in time to detection (DTD) of cultures of blood samples obtained simultaneously from a peripheral vein (PV) and from the CVC for differentiation of CRI and non-CRI. During a 15-month period, 9 episodes were categorized as CRI and 24 as non-CRI. The median DTD for patients with CRI was significantly higher than that for patients with non-CRI (457 vs. -4 min; P<.001). The optimum cutoff point for diagnosis of CRI was a DTD of > or =120 min (sensitivity, 88.9%; specificity, 100%). With pretest probability of CRI ranging from 28% to 54%, the positive predictive value of a DTD of > or =120 min for the diagnosis of CRI was 100%; the negative predictive value was 89%-96%. On the basis of findings from this study, which is the largest, to date, to involve pediatric patients with tunneled CVCs and the first to use paired quantitative blood cultures as a "criterion standard," DTD was found to be a simple, reliable tool for diagnosis of CRI in hospitals that use continuously read blood culture systems. PMID- 12905130 TI - The protease inhibitor lopinavir-ritonavir may produce opiate withdrawal in methadone-maintained patients. AB - This study examines the pharmacokinetic/pharmacodynamic interactions between (1) lopinavir-ritonavir (L/R), a fixed combination of protease inhibitors used for the treatment of HIV disease, and (2) ritonavir alone at the same dosage as that in the L/R formulation, with methadone, an opiate frequently used in substance abuse pharmacotherapy for opioid (heroin)-dependent injection drug users, many of whom are infected with HIV. L/R was associated with significant reductions in the methadone area under the concentration-time curve (P<.001), maximum concentration (P<.001), and minimum concentration (P<.001), as well as increased methadone oral clearance (P<.001) and increased opiate withdrawal symptoms (P=.013), whereas ritonavir use alone modestly and nonsignificantly increased methadone concentrations. Lopinavir is a potent inducer of methadone metabolism, and treatment with L/R requires clinical monitoring and increased methadone doses in some patients, whereas ritonavir has no significant effect on methadone metabolism. PMID- 12905131 TI - Clinical diagnosis of Legionella pneumonia revisited: evaluation of the Community Based Pneumonia Incidence Study Group scoring system. AB - This prospective case-control study sought to identify differences in presentation between Legionella pneumonia (LP) diagnosed by urinary antigen and bacteremic pneumococcal pneumonia (PP), with the aim of assessing the ability of physicians to recognize such differences at admission and validating the Community-Based Pneumonia Incidence Study (CBPIS) Group scoring system for LP diagnosis. Significant differences in presentation were found: male sex, previous receipt of beta-lactam therapy, and temperature >39 degrees C were positively associated with LP; purulent sputum, pleuritic chest pain, and previous upper respiratory tract infection were negatively associated with LP. Physicians considered Legionella to be the most likely diagnosis in 52 (64%) of 81 LP cases and in 8 (6%) of 136 PP cases. Initial administration of a macrolide and rifampin and requests for urinary antigen testing for Legionella at admission were significantly more frequent among patients with LP. Overall, the CBPIS score did not differentiate reliably between LP and PP. Although certain presenting clinical features may allow recognition of LP, it is difficult to express them in a reliable scoring system. PMID- 12905132 TI - Human cervicofacial actinomycoses: microbiological data for 1997 cases. AB - Actinomycoses are sporadically occurring endogenous polymicrobial inflammatory processes, in which fermentative actinomycetes of the genera Actinomyces, Propionibacterium, or Bifidobacterium act as the principal pathogens. Difficulties in diagnosing the disease in a timely and reliable fashion have led clinicians and microbiologists to grossly underestimate its medical importance. Therefore, we evaluated microbiological and selected clinical data derived from 1997 culture-positive cases of human cervicofacial actinomycoses examined in our laboratories during 1972-1999. The causative actinomycetes belonged to at least 9 different species, among which Actinomyces israelii and Actinomyces gerencseriae predominated. The well-known predisposition of male patients to acquire the disease varied with age and appeared to be especially pronounced in patients aged 20-60 years, the highest incidence being found in female patients aged 11-40 years and in male patients aged 21-50 years. The relevant procedures necessary for diagnosing human actinomycoses reliably, as well as details of their complex etiology, are discussed. PMID- 12905133 TI - The inventory of antibiotics in Russian home medicine cabinets. AB - The objective of this study was to inventory the stock of antimicrobials in the home medicine cabinets (HMCs) of the general population in Russia and to find out for which indications people report that they would use antibiotics without a physician's recommendation. The research was performed in 9 Russian cities by physicians who visited households. An inventory of antibiotics in HMCs was made, and respondents were asked about instances in which they would choose automedication with antibiotics. We found that 83.6% of families had antibiotics for systemic use in HMCs. The most common antibiotics in HMCs were trimethoprim sulfamethoxazole (46.3% of HMCs), ampicillin (45.1%), chloramphenicol (32.7%), erythromycin (25.5%), and tetracycline (21.8%). The major indications for automedication with antibiotics were acute viral respiratory tract infections (12.3% of total indications), cough (11.8%), intestinal disorders (11.3%), fever (9%), and sore throat (6.8%). According to this study, antibiotics are widely stocked among the general population in Russia, and people use antibiotics in an uncontrolled and imprudent manner. PMID- 12905134 TI - Aeromonas infection of the hepatobiliary system: report of 15 cases and review of the literature. AB - Aeromonas species cause both intestinal and extraintestinal disease. We reviewed hospital laboratory and medical records to identify patients with Aeromonas infection of the hepatobiliary or pancreatic system. Analysis of data from our hospital, as well as a review of the published literature, yielded a total of 41 episodes in 39 patients, and the features of these episodes are described. The most common manifestation of Aeromonas hepatobiliary infection among all reported cases was cholangitis (29 of 41 episodes). The majority of infections in our hospital occurred in patients with underlying immunosuppression or malignancy (13 of 15 patients), including 4 liver transplant recipients, and nosocomial infection was not infrequent (8 of 17 episodes). Infection occurred most commonly in patients with obstruction of the biliary tract due to stones, tumor, or stricture and was associated with a relatively high mortality rate (11.8%). Antibiotic susceptibility testing revealed that gentamicin, imipenem, and ciprofloxacin had the highest activity against the Aeromonas species isolated. PMID- 12905135 TI - Association of hepatitis C virus infection with sexual exposure in southern India. AB - To determine the association between sexual exposure and hepatitis C virus (HCV) infection in urban Chennai, India, a random sample of adults who live in a slum community completed interviews and provided samples to test for HCV, herpes simplex virus type 2 (HSV-2), and other sexually transmitted infections (STIs). All analyses excluded recent and current injection drug users. HCV infection was not associated with the reported number of sex partners for men or women. Women were more likely to be HCV infected if they reported previous genital ulcer disease (adjusted odds ratio [AOR], 3.88; 95% confidence interval [95% CI], 0.94 16.0; marginally statistically significant). Men were more likely to be HCV infected if they were HSV-2 infected (AOR, 3.85; 95% CI, 1.18-12.6) or reported having had sex with men (AOR, 3.61; 95% CI, 1.00-13.1). Sexual transmission of HCV infection may be facilitated by ulcerative STIs and male-male sexual practices, but it appears to occur infrequently in this population. PMID- 12905136 TI - Immune response to invasive Kingella kingae infections, age-related incidence of disease, and levels of antibody to outer-membrane proteins. AB - The immune response to Kingella kingae was determined by enzyme-linked immunosorbent assay, using outer-membrane proteins as coating antigen, in 19 children with invasive infection. The age-related incidence of K. kingae disease in southern Israel during 1988-2002 was calculated and correlated with serum antibody levels in healthy children. Significant increases in immunoglobulin G (IgG) levels were found in children convalescing after invasive infections. The incidence was 1.3, 40.3, 23.9, 5.7, and 1.9 cases/100,000 children among those aged 0-5, 6-11, 12-23, 24-35, and 36-47 months, respectively. A low attack rate and undetectable serum IgA and high IgG levels were found during the first 6 months of life, which indicates that protection was conferred by maternally derived immunity. The high attack rate found among 6-24-month-old children coincides with the age at which antibody levels were lowest. Low incidence of disease and increasing antibody levels were found among older children, which probably represents cumulative experience with K. kingae antigens via colonization or infection. PMID- 12905137 TI - Asymptomatic infection with Borrelia burgdorferi. AB - The natural history of asymptomatic seroconversion to Borrelia burgdorferi has been unclear. We report here, on the basis of a post hoc assessment, the frequency and outcome of asymptomatic seroconversion to B. burgdorferi in participants of a large Lyme disease vaccine trial. We show that infection with B. burgdorferi may be asymptomatic but that asymptomatic infection is unusual in the United States. PMID- 12905138 TI - Hematologic disorders associated with hepatitis C virus infection and their management. AB - More than 4 million people in the United States are infected with hepatitis C virus (HCV). During the next 20-30 years, the burden of HCV-related mortality and morbidity will likely double. To date, the most effective treatment for chronic HCV infection is the combination of either interferon (IFN)-alpha or pegylated IFN-alpha and ribavirin. For a sustained virologic response, treatment adherence and dose maintenance are essential. However, both IFN-alpha and ribavirin induce hematologic toxicity, such as anemia, neutropenia, and thrombocytopenia, which can compromise treatment adherence and dose maintenance and could, therefore, potentially influence outcomes. Although there are currently no approved treatments for hematologic complications of HCV therapy, studies have shown that hematopoietic growth factors can provide significant benefits. This review highlights the pharmacology, risks, and benefits of recombinant hematopoietic growth factor therapy in HCV-infected patients. PMID- 12905139 TI - The epidemiology of burn wound infections: then and now. AB - Burn wound infections are a serious complication of thermal injury. Although pneumonia is now the most important infection in patients with burns, burn wound infection remains a serious complication unique to the burn recipient. The methods for managing thermal injury have evolved during the past 50 years. This evolution has been accompanied by changes in the etiology, epidemiology, and approach to prevention of burn wound infections. In the 1950s, 1960s, and 1970s and into the mid-1980s, burn wounds were treated by the exposure method, with application of topical antimicrobials to the burn wound surface and gradual debridement with immersion hydrotherapy. As early burn wound excision and wound closure became the focal point of burn wound management, accompanied by a change from immersion hydrotherapy to showering hydrotherapy, the rate of burn wound infection appeared to decrease. Few epidemiologic studies have been done since this change in the approach to management of thermal injury. There are few data on the epidemiology of burn wound infections from the era of early excision and closure. Data are needed on infection rates for excised and closed burn wounds, the etiologies of these infections, and the epidemiology and the prevention of such infections. Additional studies are needed on the indications for topical and antimicrobial prophylaxis and selective decontamination of the digestive tract. PMID- 12905140 TI - Can immune markers predict subsequent discordance between immunologic and virologic responses to antiretroviral therapy? Adult AIDS Clinical Trials Group. AB - It is unclear why discordant immunologic and virologic responses occur during therapy for human immunodeficiency virus (HIV) infection. This study examined whether markers of immune activation and naive/memory lymphocyte subsets at study baseline could predict discordance between HIV type 1 (HIV-1) RNA and CD4+ lymphocyte responses at week 24 of antiretroviral therapy. Ten diverse, prospective antiretroviral studies with 1007 evaluable subjects were included. Subsets of subjects at increased risk for discordance were identified by recursive partitioning. The strongest predictor of more-favorable immunologic than virologic responses was a lower baseline CD4+ lymphocyte count. Weaker predictors in small subsets of subjects were fewer activated CD4+ lymphocytes and fewer CD8+ lymphocytes. Conversely, the strongest predictors of more-favorable virologic than immunologic responses were higher baseline CD4+ lymphocyte count and percentage. Additional predictors in some analyses were higher CD8+ lymphocyte count or percentage and lower HIV-1 RNA concentrations. Baseline markers of immune activation and naive/memory lymphocyte subsets had limited ability to predict subsequent discordance. PMID- 12905141 TI - Prevalence of Bartonella infection among human immunodeficiency virus-infected patients with fever. AB - Bartonella infection can be difficult to diagnose, especially when it manifests as bacteremia, which is usually accompanied by nonspecific symptoms, such as fever. Therefore, we hypothesized that Bartonella infection represents an underrecognized cause of febrile illness. To determine the prevalence of Bartonella infection among patients presenting with fever, we evaluated 382 patients in San Francisco. Overall, 68 patients (18%) had evidence of Bartonella infection detected by culture, indirect fluorescent antibody testing, or polymerase chain reaction (PCR). Twelve patients (3%) had either Bartonella henselae or Bartonella quintana isolated from specimens of blood, tissue, or both or had DNA detected in tissue; all 12 had concomitant human immunodeficiency virus (HIV) infection. Bartonella antibodies were detected in 17% of febrile patients, including 75% of culture-positive or PCR-positive patients. In a nested, matched case-control study aimed at identifying clinical features of febrile illness associated with Bartonella infection, only bacillary angiomatosis and elevated alkaline phosphatase levels were associated with Bartonella infection (P< or =.03 for both). The prevalence of Bartonella infection among patients with late-stage HIV infection and unexplained fever is much greater than has previously been documented. PMID- 12905142 TI - Cytomegalovirus (CMV) and human immunodeficiency virus (HIV) burden, CMV end organ disease, and survival in subjects with advanced HIV infection (AIDS Clinical Trials Group Protocol 360). AB - We undertook a prospective study to analyze cytomegalovirus (CMV) end-organ disease (EOD) in subjects with advanced human immunodeficiency virus (HIV) infection. Of 403 individuals without prior CMV EOD who were followed up for a median of 151 weeks, 56 died and 21 developed CMV EOD. Twenty of the subjects with CMV EOD had CD4 cell counts of < or =50 cells/mm3 and HIV RNA level of >10,000 copies/mL of plasma at baseline; in these 20 subjects, an increase of CMV DNA level to greater than the quantification limits was associated with CMV EOD. A CD4 cell count of < or =100 cells/mm3 and an HIV RNA level of >10,000 copies/mL of plasma at baseline, a CMV DNA level of >200 copies/mL of blood during follow up, or development of CMV EOD were all associated with decreased survival. HIV infected subjects with CD4 cell counts of < or =50 cells/mm3 and HIV RNA levels of >10,000 copies/mL of plasma should have blood fractions screened for CMV DNA; if CMV DNA is detected, CMV prophylaxis might be considered. PMID- 12905143 TI - Increased prevalence of hypothyroidism among human immunodeficiency virus infected patients: a need for screening. AB - The aim of this cross-sectional multicenter study was to determine the prevalence of and risk factors for hypothyroidism in human immunodeficiency virus (HIV) infected patients. Free T4, free T3, and thyroid-stimulating hormone levels were determined. Data on age, sex, weight variation, smoking status, duration of HIV infection, Centers for Disease Control and Prevention disease stage, CD4 cell count, HIV RNA load, lipodystrophy, HIV-hepatitis C virus coinfection, and antiretroviral treatment (type of drugs and total cumulative dose) were collected. The prevalence study included 350 HIV-infected patients. Sixteen percent of patients had hypothyroidism: 2.6% had overt hypothyroidism, 6.6% had subclinical hypothyroidism, and 6.8% had a low free T4 level. The prevalence of subclinical hypothyroidism was higher among HIV-infected men than among HIV infected women. A case-control study was conducted that compared hypothyroid (n=56) and euthyroid (n=287) patients. In the multivariate analysis, receipt of stavudine and low CD4 cell count were associated with hypothyroidism. Therefore, screening may be indicated for patients, especially men, who have received stavudine or have decreased CD4 cell counts. PMID- 12905144 TI - Epidemiological, clinical, and prognostic differences between the diseases caused by Mycobacterium kansasii and Mycobacterium tuberculosis in patients infected with human immunodeficiency virus: a multicenter study. AB - A multicenter, comparative study was performed to determine the epidemiological, clinical, and prognostic differences between the diseases caused by Mycobacterium tuberculosis and Mycobacterium kansasii in human immunodeficiency virus (HIV) infected patients. From 1 January 1995 through 31 December 1999, 25 HIV-infected patients received diagnoses of M. kansasii infection, and another 75 were selected as control subjects from among patients who had M. tuberculosis infection. Variables associated with M. tuberculosis disease in the multivariate analysis were previous intravenous drug use (odds ratio [OR], 8; 95% confidence interval [CI], 1.5-41.4) and interstitial radiologic pattern (OR, 12.7; 95% CI, 1.7-94.3). Variables associated with M. kansasii were previous diagnosis of acquired immunodeficiency syndrome (OR, 15.8; 95% CI, 4.2-59.6) and concomitant opportunistic infections (OR, 14.2; 95% CI, 2-105.7). Clinical and radiologic features were similar for both groups, but epidemiological characteristics and prognosis were different. M. kansasii disease was associated more closely with level of immunosuppression and progression of HIV infection than was disease caused by M. tuberculosis. PMID- 12905145 TI - Role of hepatitis B virus genotypes in chronic hepatitis B exacerbation. AB - Hepatitis B virus (HBV) genotypes and precore and core promoter mutations were determined in 318 patients with HBV. Patients infected with HBV genotype B had a higher median alanine aminotransferase level and bilirubin level and a lower median albumin level during exacerbations of disease, compared with patients infected with HBV genotype C (all P<.001). By logistic regression analysis, HBV genotype B infection (P=.014) and low albumin levels (P=.006) were independently associated with a higher risk of hepatic decompensation during severe exacerbations of disease. Patients infected with genotype B had a significantly higher mortality due to hepatic decompensation than did patients with genotype C (70% vs. 27.8%; P=.05). PMID- 12905146 TI - Fatal human rabies caused by European bat Lyssavirus type 2a infection in Scotland. AB - We wish to report the first recorded case of indigenous human rabies caused by a bat bite in the United Kingdom in 100 years. This instructive case report highlights a number of key lessons: first, bites from insectivorous bats indiginous to the United Kingdom can cause rabies in humans; second, rabies immunization is essential for bat-handlers, and postexposure treatment for rabies is essential for patients bitten by bats; third, patients able to give a history who present with acute flaccid paralysis and/or presumptive viral encephalitis should be asked if they have been bitten by bats, irrespective of travel history, or this history should be obtained from family or friends; fourth, antemortem diagnosis of bat rabies (EBLV type 2a infection) in humans is possible using RT PCR. PMID- 12905147 TI - Detection and quantification of Mycoplasma genitalium in male patients with urethritis. AB - Detection and quantification of Mycoplasma genitalium were evaluated in 83 patients with urethritis (group 1), 60 patients with urethral symptoms but no urethritis (group 2), and 50 asymptomatic men (group 3). Quantification of M. genitalium was carried out using real-time polymerase chain reaction (PCR) analysis of first-pass urine samples. The rate of detection of M. genitalium was significantly higher in group 1 than in groups 2 and 3 (P<.0001). The mean observed concentration of M. genitalium was 1.2x10(4) equivalent genomes/mL of urine (range, 50 to 8x10(4) equivalent genomes/mL). Analysis of M. genitalium load in serial urine samples collected before and after the administration of antibacterial treatment showed an association between clinical and microbiological responses, with a shift to negative PCR results in symptom-free patients. Our results illustrate the usefulness of monitoring the M. genitalium load in evaluating the susceptibility of M. genitalium to antibacterial treatment. PMID- 12905148 TI - Human herpesvirus 8-associated diseases in solid-organ transplantation: importance of viral transmission from the donor. PMID- 12905149 TI - Rifamycin treatment of tuberculosis in a patient receiving atenolol: less interaction with rifabutin than with rifampin. PMID- 12905150 TI - Wiggling subcutaneous lumps. PMID- 12905151 TI - Cytomegalovirus-induced gingival hyperplasia. AB - We report a rare presentation of gingival hyperplasia associated with cytomegalovirus infection in a 33-year-old human immunodeficiency virus-infected man. PMID- 12905152 TI - Clinical review of nontyphoid Salmonella infections from 1991 to 1999 in a Danish county. AB - To determine outcomes of nontyphoid Salmonella infections, we reviewed all 3328 cases of infection registered in a large Danish county from 1991 to 1999. The hospitalization rate was 27% among patients <5 years old, 22% among patients 5-59 years old, and 54% among patients > or =60 years old; complications were present in 7% and extraintestinal disease in 4% of patients. We conclude that nontyphoid Salmonella infections may frequently cause severe disease even among patients without predisposing conditions. PMID- 12905153 TI - Inaccurate glycosylated hemoglobin A1C measurements in human immunodeficiency virus-positive patients with diabetes mellitus. AB - Persistent differences in blood glucose and serum glycosylated hemoglobin (HbA1C) measurements were observed in 4 human immunodeficiency virus-positive patients with diabetes mellitus, all of whom were taking drugs associated with hemolysis, which interferes with the reliability of HbA1C levels. Determination of fructosamine levels was a more accurate alternative for measuring average glycemic control in these patients. PMID- 12905434 TI - Managing the trauma pandemic: learning from the past. PMID- 12905154 TI - Miller-Dieker syndrome: analysis of a human contiguous gene syndrome in the mouse. PMID- 12905435 TI - Treatment considerations for comminuted mandibular fractures. AB - PURPOSE: We sought to assess the methods of treatment used and outcomes for a large sample of patients with comminuted fractures of the mandible. PATIENTS AND METHODS: The records of all patients who were identified as having comminuted fractures of the mandible over a 10-year period with sufficient follow-up were collected and analyzed for demographic information, treatment rendered, and outcomes. Standard descriptive statistics and nonparametric statistics were used to analyze the data. RESULTS: A total of 196 patients (167 males and 29 females) with 198 comminuted fractures met the inclusion criteria. The mandibular body was the most commonly affected region. Approximately half were sustained in altercations. Gunshot wounds created fractures that were more comminuted than other causes. The comminuted regions were treated by closed reduction and maxillomandibular fixation (MMF) in 35 fractures, open reduction with stable internal fixation in 146 fractures, and 17 were treated with external pin fixation. For those patients treated with open reduction, a single reconstruction bone plate was used in the majority of cases (114). For those patients treated open, 98 were treated using an intraoral approach and 52 were treated using an extraoral approach. The mean follow-up was 140.6 days. Complications occurred in 26 fractures (13%). The complications were malocclusion in 8 fractures and nonocclusal (ie, infection, nonunion, etc) in 18 fractures. There was a statistically significant relationship between the development of complications and the degree of fragmentation (P <.05). There was also a significant relationship between treatment and the development of complications (P <.05). Patients treated with external pin fixation had a 35.2% complication rate compared with a 17.1% complication rate for patients undergoing closed treatment with MMF, or patients treated with open reduction and stable internal fixation (10.3%). However, patients treated with external pin fixation had more severe injuries. CONCLUSIONS: The results of this study show that, when possible, the use of open reduction and stable internal fixation is associated with a low complication rate. However, not all comminuted fractures are amenable to this treatment, and in those, alternatives such as closed reduction with MMF or the application of external pin fixation may be necessary. PMID- 12905436 TI - A clinical evaluation of dry socket in a Nigerian teaching hospital. AB - PURPOSE: We sought to determine the incidence of dry socket in a Nigerian teaching hospital and to evaluate the patients' demographic pattern, predisposing factors, the treatment given, and treatment outcome. PATIENTS AND METHODS: A retrospective review of records of dental extractions complicated by dry socket in Obafemi Awolowo University Teaching Hospital, Ile-Ife, between January 1996 and December 2000 was undertaken. Information retrieved included patient sociodemographic data, indications for extraction, tooth extracted, status of attending surgeon, onset of symptoms, relevant findings of the examining clinician, interval before presentation, treatment given, and its outcome. RESULTS: Of the 3,319 dental extractions performed in 3,008 patients, 136 (4.1%) were complicated by dry socket. The patients' mean age was 33.4 (15.4) years and a peak age incidence of 21 to 30 years was found. A slight female preponderance (1.4:1) was observed. The majority of patients were in the low-income group, and presentation in the hospital was prompt in the high-income group. Mandibular teeth were affected 3 times more than maxillary teeth (P =.00080). Most cases of dry socket resulted from extractions performed by undergraduates and house officers. Various underlying systemic conditions were found in 11.0% of cases, none of which included use of oral contraceptives. Treatment was usually the use of zinc oxide eugenol dressing in an irrigated socket, combined with antibiotic therapy in 45.3% of cases. No adverse reaction to zinc oxide eugenol was observed. CONCLUSION: The incidence of dry socket in our hospital is 4.1%. The mandible was involved 3 times more than the maxilla. With the use of zinc oxide eugenol dressing, 70.6% of patients completed treatment satisfactorily and 29.2% were lost to follow-up. PMID- 12905438 TI - Le Fort III osteotomy: a new internal positioned distractor. AB - PURPOSE: The treatment of midfacial retrusion, due to congenital midfacial hypoplasia or trauma, is a challenging problem. Conventional Le Fort III osteotomies, with the surgical mobilisation of the entire midface, offers one solution. Distraction osteogenesis has become an accepted, and often preferred, technique for the treatment of severe midfacial hypoplasia. Existing distraction devices have several disadvantages. To overcome some of these disadvantages, an internal and temporal positioned distraction device was developed. PATIENTS AND METHODS: The new, internal positioned distractor device was used in 5 patients (age range, 8 to 15 years). Nine distractors were used in 4 patients with midfacial retrusion and 1 patient with hemifacial microsomia. RESULTS: The preoperative planned position of the osteotomized Le Fort III segment was obtained without any complications in all cases. Midface advancement, measured as the anterior displacement of the infraorbital rim, ranged from 14 to 20 mm. The distractor was almost invisible and did not disturb daily activities of the patient. Activation of the distractor was performed by rotation of a small lateral activation rod. CONCLUSION: These preliminary results encourage the further use of the new distractor device in a larger series of patients. PMID- 12905437 TI - Condylar erosion and disc displacement: detection with high-resolution ultrasonography. AB - PURPOSE: Our goal was to determine the feasibility of high-resolution sonography for the detection of condylar erosion and associated disc displacement at the temporomandibular joint (TMJ) condyle. MATERIALS AND METHODS: Forty-eight consecutive patients (96 joints) with TMJ disorders were investigated prospectively using a 12.5-MHz array transducer. Images were assessed for condylar erosion and disc displacements. Sonographic findings were correlated with those of magnetic resonance imaging (MRI). RESULTS: At MRI, 18 osseous changes were diagnosed with condylar erosion. Sonographically, 15 of the 18 condylar erosions were diagnosed correctly. Sensitivity, specificity, and accuracy of sonography in the depiction of condylar erosion were 83%, 63%, and 67%, respectively. The positive predictive value was 34%, and the negative predictive value was 94%. Disc displacement without reduction (43 of 96) and disc displacement without reduction concomitant with condylar erosion (16 of 96) were detected with an accuracy of 93% and 80%. CONCLUSION: Sonography is an insufficient imaging technique for the detection of condylar erosion. Assessment of disc displacement without reduction may be reliably made with sonography. PMID- 12905439 TI - Frozen section examination of the margins for resection of squamous cell carcinoma of the lower lip. AB - OBJECTIVES: Our goal was to evaluate the role of frozen section margin examination in the surgical management of squamous cell carcinoma of the lower lip. MATERIALS AND METHODS: We reviewed frozen section examination of the resection surfaces of 131 consecutive patients surgically treated for squamous cell carcinoma of the lower lip during the period January 1980 to January 1999. When invasive carcinoma or carcinoma in situ was present at the resection surface, the margin was defined as positive and re-resection was performed. RESULTS: A peroperative positive margin was seen in 18 of 131 (14%). In 8 patients (6%), a local recurrence developed. CONCLUSIONS: Frozen section examination for margin assessment in the surgical treatment of lip cancer is a reliable technique to control the radicality of the procedure; a secondary surgical procedure or postoperative radiotherapy can be reduced. However, a tumor free resection surface does not guarantee that local recurrence will not occur. PMID- 12905441 TI - Trigeminal nerve hypesthesia after sagittal split osteotomy in setback cases: correlation of postoperative computed tomography and long-term trigeminal somatosensory evoked potentials. AB - PURPOSE: We sought to assess the relationship of the inferior alveolar nerve to the osteotomy site after bilateral sagittal split osteotomy (BSSO) and to correlate the nerve osteotomy relationship as the cause of long-term postoperative hypesthesia. PATIENTS AND METHODS: The subjects consisted of 28 patients with mandibular prognathism who underwent BSSO setback surgery. The distance between the ostectomized surface of the mandibular ramus and mandibular canal was measured and scored on a computed tomograph (CT). The relationship between this distance and the presence and degree or absence of trigeminal nerve hypesthesia was objectively evaluated by latency delay of the trigeminal somatosensory evoked potential (TSEP) records. RESULTS: Five of the 56 sides (8.9%) showed latency delays more than 1 year after the operations. Scored CT points between the canal and the split surface strongly correlated with TSEP latency recovery (Y = 1.716X + 41.2). CONCLUSIONS: This relationship between the canal and the osteotomy site is related to long-term hypesthesia in BSSO postoperatively. PMID- 12905442 TI - Volumetric and histologic responses to radiotherapy or radiochemotherapy of metastatic cervical lymph nodes of oral squamous cell carcinoma. AB - PURPOSE: The relationship between the volumetric and histologic responses of metastatic cervical nodes to radiotherapy or chemotherapy in the oral and maxillofacial region is unclear. In this study, we evaluated the correlation between the initial volume and regression rate of metastatic nodes with their histologic response to preoperative radiotherapy or radiochemotherapy. PATIENTS AND METHODS: The volume of 54 metastatic nodes in 32 patients with squamous cell carcinoma in the oral and maxillofacial region was measured by ultrasonography before and after preoperative therapy, and the rate of the volume change was calculated. All surgically removed nodes were histologically classified as poor, good, or complete response according to their histologic features. RESULTS: There was no significant difference in initial volume among the 3 response groups. Good and complete response nodes showed a significant increase in regression rate compared with poor response nodes. All 11 nodes showing no regression were poor response nodes, and 7 with a regression rate of more than 90% were good or complete response nodes. The remaining 36 nodes (regression rate, 0% to 90%) represented all 3 types of histologic response. Of these, 7 of 9 complete response nodes were found in 5 patients who received combination chemotherapy consisting of 5-fluorouracil, leucovorin, and cisplatin. CONCLUSIONS: The initial nodal volume before therapy is not a good indicator for the response to radiotherapy and/or chemotherapy. A regression rate of more than 90% may be a useful predictor of the effectiveness of preoperative treatments, but it was difficult to define the cutoff values in regression rates for differentiating types of histologic response. PMID- 12905444 TI - Evaluation of 67 Ga citrate and 99m Tc bone scintigraphy at initial examination for primary oral squamous cell carcinoma. AB - PURPOSE: This study was carried out to evaluate the diagnostic utility of gallium citrate Ga 67 ((67)Ga) and technetium Tc 99m bone scintigraphy as initial screening methods for distant metastasis and synchronous malignancies in patients with oral cancer. PATIENTS AND METHODS: Ga and/or bone scintigraphy was performed for 123 patients with primary squamous cell carcinoma (SCC). Abnormal accumulation sites detected with scintigraphy were reevaluated with radiography or computed tomography. Whether the accumulation sites were metastatic was determined according to the radiographic findings and the later clinical course of the patients. RESULTS: Ga scintigraphy did not detect distant metastases or 4 synchronous cancers at other sites and provided no useful information on the emergence of delayed pulmonary metastases that developed in 3 cases after a few months. Ga scintigraphy showed an abnormal accumulation in 53% of primary tumors and 25% of metastatic cervical lymph nodes, and the detection of accumulation was closely related to the size of the lesion. Bone scintigraphy showed 1 case (0.9%) with distant metastasis to the vertebra; however, there were numerous false positive cases that required additional radiographic confirmation. CONCLUSIONS: These results suggested that Ga and bone scintigraphy at the initial examination of patients with primary oral SCC are ineffective for detecting distant metastasis and other malignancies because of the low incidence of these lesions and the diagnostic inaccuracy of scintigraphy. PMID- 12905443 TI - Effects of cerulenin on the endogenous fatty acid synthetic activity in squamous cell carcinoma of the oral cavity. AB - PURPOSES: Inhibition of cerulenin on the endogenous fatty acid synthetic activities of oral squamous cell carcinoma (OSCC) and normal oral mucosa was assayed. METHODS: Squamous cell carcinoma and normal oral mucosa were collected fresh from surgical specimens. The collected tissues were minced in RPMI 1640 and divided into 3 groups: cerulenin treated, dimethylsulfoxide treated, and control. The tissues were incubated in [1(2)-(14)C]acetic acid, sodium salt for the last 2.5 hours of the treatment at 37 degrees C in 5% CO(2). After labeling, total lipids were extracted and counted for (14)C by scintillation counting. RESULTS: Endogenous fatty acid synthetic activities of oral squamous cell caranoma in the cerulenin-treated group decreased by 19% at 1 hour, 64% at 2 hours, and 87% at 4 hours; remained nearly unchanged in the dimethylsulfoxide-treated group; and increased slightly in the control group. The oral mucosa tissues were only mildly affected by cerulenin in fatty acid synthesis. CONCLUSIONS: Cerulenin significantly inhibits fatty acid synthetic activity in squamous cell carcinoma and only mildly affected the oral mucosa, indicating that the fatty acid synthetic pathway may be exploited as a target for developing anticancer drugs. PMID- 12905445 TI - Alterations of morphology and microdensity in the condyle after mandibular osteodistraction in the rat. AB - PURPOSE: In this study, we examined the effects of mandibular distraction osteogenesis on the morphology and the microdensity of the rat condyle. MATERIALS AND METHODS: One hundred twenty-nine rats were allocated to 4 experimental groups (n = 32 or 33). Each received unilateral mandibular ramus osteotomy and distraction device placement. After a 3-day latency, these were distracted once a day for 5 days. The slow distraction group was distracted a total of 1 mm (0.2 mm/d); the moderate group, 2 mm (0.4 mm/d); the rapid group, 3 mm (0.6 mm/d); and the sham group, no distraction (0.0 mm/d). Eight to 9 rats in each group were sacrificed at each of 4 time points after device placement (6, 10, 24, and 38 days). Baseline data were obtained 3 days after osteotomy and device placement without distraction from an additional 10 rats. Radiographs of the hemimandibles were scanned and measured to evaluate changes in condylar size (height, width, and area), angulation, and bone microdensity converted to equivalent bone thickness using a stepwedge. The wet weights of masseter muscle were measured at the time of harvest. RESULTS: 1) Muscle weight gains over time were significantly lower in the treated than the untreated sides throughout the consolidation period (P <.001). 2) Condylar size and angulation on the untreated side increased postoperatively, whereas there was a significant reduction of these parameters (P <.01 to.001) on the treated side at 24 and/or 38 days. 3) Condylar microdensity significantly increased on the untreated side at 24 and 38 days (P <.05 to.01) but not on the treated side. 4) Faster or larger distraction caused more severe size reduction and more upright condylar angulation, prevented an increase in bone microdensity on the treated side, especially during the consolidation periods (P <.05 to.01), and retarded increase in muscle weight, whereas a slower distraction rate showed few negative, and even some positive effects. 5) Correlations in size, angulation, and microdensity between right and left condyles became less significant over time. 6) There were positive correlations between muscle weight and condylar size, angulation, and microdensity. CONCLUSION: An increased rate of mandibular distraction has significant negative effects on condylar morphology and microdensity. PMID- 12905446 TI - A review of the class III antiarrhythmic agents and their uses and implications for patient management. PMID- 12905447 TI - Firearm injuries to the maxillofacial region: an overview of current thoughts regarding demographics, pathophysiology, and management. PMID- 12905448 TI - Maxillofacial surgery in World War I: the role of the dentists and surgeons. PMID- 12905449 TI - A case report of submandibular aggressive fibromatosis in a 16-month-old infant with 8-year postsurgical follow-up. PMID- 12905450 TI - An unusual complication of radial artery cannulation and its management: a case report. PMID- 12905451 TI - Massive osteolysis of the mandible: a case report. PMID- 12905452 TI - Rhabdomyolysis in the acutely cocaine- intoxicated patient sustaining maxillofacial trauma: report of a case and review of the literature. PMID- 12905453 TI - Lemierre's syndrome: a case report. PMID- 12905454 TI - Use of a modified distraction appliance to treat radiation-induced trismus. PMID- 12905459 TI - Tuberculosis in the cytokine era: what rheumatologists need to know. PMID- 12905460 TI - The JAK/STAT pathway in rheumatoid arthritis: pathogenic or protective? PMID- 12905461 TI - What is "resistance" in Paget's disease of bone? PMID- 12905462 TI - Sex hormones and systemic lupus erythematosus: review and meta-analysis. PMID- 12905463 TI - How do antiphospholipid antibodies bind beta2-glycoprotein I? PMID- 12905464 TI - Treatment of rheumatoid arthritis with tumor necrosis factor inhibitors may predispose to significant increase in tuberculosis risk: a multicenter active surveillance report. AB - OBJECTIVE: The long-term safety of therapeutic agents that neutralize tumor necrosis factor (TNF) is uncertain. Recent evidence based on spontaneous reporting shows an association with active tuberculosis (TB). We undertook this study to determine and describe the long-term safety of 2 of these agents, infliximab and etanercept, in rheumatic diseases based on a national active surveillance system following the commercialization of the drugs. METHODS: We analyzed the safety data actively collected in the BIOBADASER (Base de Datos de Productos Biologicos de la Sociedad Espanola de Reumatologia) database, which was launched in February 2000 by the Spanish Society of Rheumatology. For the estimation of TB risk, the annual incidence rate in patients treated with these agents was compared with the background rate and with the rate in a cohort of patients with rheumatoid arthritis (RA) assembled before the era of anti-TNF treatment. RESULTS: Seventy-one participating centers sent data on 1,578 treatments with infliximab (86%) or etanercept (14%) in 1,540 patients. Drug survival rates (reported as the cumulative percentage of patients still receiving medication) for infliximab and etanercept pooled together were 85% and 81% at 1 year and 2 years, respectively. Instances of discontinuation were essentially due to adverse events. Seventeen cases of TB were found in patients treated with infliximab. The estimated incidence of TB associated with infliximab in RA patients was 1,893 per 100,000 in the year 2000 and 1,113 per 100,000 in the year 2001. These findings represent a significant increased risk compared with background rates. In the first 5 months of 2002, after official guidelines were established for TB prevention in patients treated with biologics, only 1 new TB case was registered (in January). CONCLUSION: Therapy with infliximab is associated with an increased risk of active TB. Proper measures are needed to prevent and manage this adverse event. PMID- 12905465 TI - New radiographic bone erosions in the wrists of patients with rheumatoid arthritis are detectable with magnetic resonance imaging a median of two years earlier. AB - OBJECTIVE: In a 5-year followup study, we investigated the temporal relationship between development of wrist joint erosions as visualized by magnetic resonance imaging (MRI) versus conventional radiography (CR), in patients with rheumatoid arthritis. We also evaluated the risk of erosive progression on CR associated with the presence of MRI erosions. METHODS: In 10 patients with rheumatoid arthritis, MRI and CR of the dominant wrist were performed annually for 5 years. In each image set, each wrist bone (metacarpal bases, carpal bones, radius, and ulna) was assessed for the absence or presence of bone erosions. RESULTS: Nine bones showed radiographic erosions at baseline. Twenty-seven new radiographic erosions developed during the 5-year followup period. Of these 27 new erosions, 21 were detected 1-5 years earlier by MRI than by CR, 3 were simultaneously detected by both methods, 2 were detected 1-2 years later by MRI than by CR, and 1 erosion (radiographically detected at 5-year followup) was not visualized with MRI. MRI detection of new radiographic erosions preceded CR detection by a median of 2 years. In bones with MRI erosions at baseline, the relative risk of radiographic erosions at 5-year followup was 4.5 (95% confidence interval [95% CI] 2.6-7.6), compared with bones without baseline MRI erosions. If bones with baseline radiographic erosions were excluded from the analysis, the relative risk was 4.1 (95% CI 2.2-7.5). CONCLUSION: Most new radiographic bone erosions (78%) were visualized at least 1 year earlier by MRI than by CR. This illustrates that the information on joint destruction provided by CR is considerably delayed compared with that provided by MRI. A significantly increased risk of progression of radiographic erosion in bones with baseline MRI erosions was observed, demonstrating a prognostic value of MRI with respect to long-term radiographic outcome. PMID- 12905466 TI - Rheumatoid arthritis is a heterogeneous disease: evidence for differences in the activation of the STAT-1 pathway between rheumatoid tissues. AB - OBJECTIVE: To generate a molecular description of synovial tissue from rheumatoid arthritis (RA) patients that would allow us to unravel novel aspects of pathogenesis and to identify different forms of disease. METHODS: We applied complementary DNA microarray analysis to profile gene expression, with a focus on immune-related genes, in affected joint tissues from RA patients and in tissues from osteoarthritis (OA) patients as a control. To validate microarray data, real time polymerase chain reaction was performed on genes of interest. RESULTS: The gene expression signatures of synovial tissues from RA patients showed considerable variability, resulting in the identification of at least two molecularly distinct forms of RA tissues. One class of tissues revealed abundant expression of clusters of genes indicative of an involvement of the adaptive immune response. Detailed analysis of the expression profile provided evidence for a prominent role of an activated signal transducer and activator of transcription 1 pathway in these tissues. The expression profiles of another group of RA tissues revealed an increased tissue remodeling activity and a low inflammatory gene expression signature. The gene expression pattern in the latter tissues was reminiscent of that observed in the majority of OA tissues. CONCLUSION: The differences in the gene expression profiles provide a unique perspective for distinguishing different pathogenetic RA subsets based on molecular criteria. These data reflect important aspects of molecular variation that are relevant for understanding the biologic dysregulation underlying these subsets of RA. This approach may also help to define homogeneous groups for clinical studies and evaluation of targeted therapies. PMID- 12905468 TI - Tumor necrosis factor alpha blockade reduces the synovial cell infiltrate early after initiation of treatment, but apparently not by induction of apoptosis in synovial tissue. AB - OBJECTIVE: To determine whether treatment with the chimeric anti-tumor necrosis factor alpha antibody infliximab could reduce cellularity by the induction of apoptosis in synovial tissue. METHODS: Twenty-four rheumatoid arthritis patients with active disease were randomized to receive either infliximab (3 mg/kg) (n = 12) or placebo (n = 12) intravenously. All patients were subjected to arthroscopic synovial biopsy directly before initiation of treatment. A second arthroscopic synovial biopsy of the same index joint was performed 48 hours after the first arthroscopy. After the second arthroscopy, the patients who had initially received placebo were also treated with infliximab in an extension study. A third arthroscopy was performed in all patients on day 28. Immunohistologic analysis was performed to characterize the cell infiltrate. In situ detection of apoptotic cells was performed by TUNEL assay and electron microscopy. RESULTS: At 48 hours after initiation of infliximab treatment, there was a significant reduction in the number of intimal macrophages; this was not observed in the placebo group. The number of sublining macrophages, T cells, and plasma cells also tended to be decreased in infliximab-treated patients, but not in the placebo group. Of interest, we did not detect any increase in the number of apoptotic cells after infliximab treatment. CONCLUSION: Infliximab therapy may reduce the number of inflammatory cells in rheumatoid synovial tissue as soon as 48 hours after initiation of treatment, but apparently not by induction of apoptosis. Conceivably, decreased cell infiltration primarily results from early inhibition of cell migration. PMID- 12905467 TI - Serologic changes following B lymphocyte depletion therapy for rheumatoid arthritis. AB - OBJECTIVE: To explore the changes in serologic variables and clinical disease activity following B lymphocyte depletion in 22 patients with rheumatoid arthritis (RA). METHODS: B lymphocyte depletion was attained using combination therapy based on the monoclonal anti-CD20 antibody rituximab. Levels of a serologic indicator of inflammation, C-reactive protein (CRP), of antimicrobial antibodies, of autoantibodies including IgA-, IgM-, and IgG-class rheumatoid factors (RF), and of antibodies to cyclic citrullinated peptide (anti-CCP) were assayed. RESULTS: The majority of patients showed a marked clinical improvement after treatment with rituximab, with benefit lasting up to 33 months. Levels of total serum immunoglobulins fell, although the mean values each remained within the normal range. Whereas the IgM-RF response paralleled the changes in total serum IgM levels, the levels of IgA-RF, IgG-RF, and IgG and anti-CCP antibodies decreased significantly more than did those of their corresponding total serum immunoglobulin classes. The kinetics for the reduction in CRP levels also paralleled the decreases in autoantibody levels. In contrast, levels of antimicrobial antibodies did not change significantly. B lymphocyte return occurred up to 21 months posttreatment. The time to relapse after B lymphocyte return was often long and unpredictable (range 0-17 months). Relapse was, however, closely correlated with rises in the level of at least one autoantibody. Increased autoantibody levels were rarely observed in the absence of clinical change. CONCLUSION: Following B lymphocyte depletion in patients with RA, a positive clinical response occurred in correlation with a significant drop in the levels of CRP and autoantibodies. Antibacterial antibody levels were relatively well maintained. B lymphocyte return preceded relapse in all patients. There was also a temporal relationship between clinical relapse and rises in autoantibody levels. Although these observations are consistent with a role for B lymphocytes in the pathogenesis of RA, the precise mechanisms involved remain unclear. PMID- 12905469 TI - Synovial fluid exoglycosidases are predictors of rheumatoid arthritis and are effective in cartilage glycosaminoglycan depletion. AB - OBJECTIVE: To analyze enzymes involved in joint damage by simultaneous investigation of glycosidases and matrix metalloproteinases (MMPs) in patients with various joint diseases. METHODS: Activities of glycosidases (beta-D glucuronidase, beta-D-N-acetyl-glucosaminidase, beta-D-N-acetyl galactosaminidase, beta-D-galactosidase, and alpha-D-mannosidase) were tested at an acidic pH as well as at the original pH of the synovial fluid (SF) samples in parallel with activities of MMP-1 and MMP-9. RESULTS: Patients with rheumatoid arthritis (RA) were characterized by significantly elevated activities of beta-D glucuronidase and beta-D-N-acetyl-glucosaminidase in SF compared with patients with osteoarthritis, seronegative spondylarthritis, or acute sports injury. To select the best predictor for distinguishing among patient groups, a stepwise logistic regression analysis was performed; the strongest association was found to be between RA and beta-D-glucuronidase/beta-D-N-acetyl-glucosaminidase activities (measured at the pH of the SF). Further, a significant correlation was observed between the activity of SF beta-D-N-acetyl-glucosaminidase and the level of rheumatoid factor. In vitro digestion of human hyaline cartilage samples revealed that the dominant glycosidases, alone or in combination with MMPs, proved to be effective in depleting glycosaminoglycans (GAGs) from cartilage. CONCLUSION: These results suggest that exoglycosidases, which are present in the SF of RA patients, may contribute to the depletion of GAGs from cartilage and thereby facilitate the invasion of synovial cells and their attachment to cartilage in RA. PMID- 12905470 TI - Inflammation and angiogenesis in osteoarthritis. AB - OBJECTIVE: To quantify the relationship between inflammation and angiogenesis in synovial tissue from patients with osteoarthritis (OA). METHODS: Hematoxylin and eosin staining and histologic grading for inflammation were performed for 104 patients who met the American College of Rheumatology criteria for OA and had undergone total joint replacement or arthroscopy. A purposive sample of synovial specimens obtained from 70 patients was used for further analysis. Vascular endothelium, endothelial cell (EC) proliferating nuclei, macrophages, and vascular endothelial growth factor (VEGF) were detected by immunohistochemical analysis. Angiogenesis (EC proliferation, EC fractional area), macrophage fractional area, and VEGF immunoreactivity were measured using computer-assisted image analysis. Double immunofluorescence histochemical analysis was used to determine the cellular localization of VEGF. Radiographic scores for joint space narrowing and osteophyte formation in the knee were also assessed. RESULTS: Synovial tissue samples from 32 (31%) of 104 patients with OA showed severe inflammation; thickened intimal lining and associated lymphoid aggregates were often observed. The EC fractional area, EC proliferation, and VEGF immunoreactivity all increased with increasing histologic inflammation grade and increasing macrophage fractional area. In the synovial intimal lining, VEGF immunoreactivity was localized to macrophages and increased with increasing EC fractional area and angiogenesis. No inflammation or angiogenic indices were significantly correlated with radiographic scores. CONCLUSION: Inflammation and angiogenesis in the synovium are associated with OA. The angiogenic growth factor VEGF generated by the inflamed synovium may promote angiogenesis, thereby contributing to inflammation in OA. PMID- 12905471 TI - Impact of type of meniscal tear on radiographic and symptomatic knee osteoarthritis: a sixteen-year followup of meniscectomy with matched controls. AB - OBJECTIVE: To investigate long-term radiographic and patient-relevant outcome of isolated limited meniscectomy with regard to type of meniscal tear and extent of surgical resection. METHODS: We studied 155 patients with intact cruciate ligaments (mean +/- SD age 54 +/- 12 years) who had undergone meniscectomy an average of 16 +/- 1 years earlier. The patients were examined using standardized radiography and validated self-administered questionnaires. The Knee Injury and Osteoarthritis Outcome Score (KOOS) was used to quantify knee-related symptoms, and the definition of a symptomatic knee was determined. We used 68 control subjects matched for age, sex, and body mass index to calculate the relative risks (RRs). RESULTS: Radiographic tibiofemoral osteoarthritis (OA) (Kellgren/Lawrence grade > or =2) was present in 66 index knees (43%), of which 39 (59%) were considered to be symptomatic according to the KOOS. In total, 77 patients (50%) had a symptomatic index knee. In a multivariate model, degenerative meniscal tears were associated with both radiographic OA (P = 0.030) and combined radiographic and symptomatic OA (P < or = 0.015). The RRs for combined radiographic and symptomatic OA after degenerative and traumatic types of meniscal tear were 7.0 (95% confidence interval [95% CI] 2.1-23.5) and 2.7 (95% CI 0.9-7.7), respectively, compared with matched controls. CONCLUSION: An isolated meniscal tear treated by limited meniscectomy is associated with a high risk of radiographic and symptomatic tibiofemoral OA at 16-year followup. Factors associated with worse outcome were degenerative meniscal lesions and extensive resections. We suggest that degenerative meniscal tears may be associated with incipient OA, and that the meniscal tear signals the first symptom of the disease. PMID- 12905472 TI - The combination of insulin-like growth factor 1 and osteogenic protein 1 promotes increased survival of and matrix synthesis by normal and osteoarthritic human articular chondrocytes. AB - OBJECTIVE: Although growth factor therapy could be an attractive method for stimulating the repair of damaged cartilage matrix, there is evidence that with aging and/or with the development of osteoarthritis (OA), articular chondrocytes may become unresponsive to growth factor stimulation. The aim of the current study was to compare the ability of insulin-like growth factor+(IGF-1) and osteogenic protein+(OP-1), alone and in combination, to stimulate human normal and OA chondrocytes in culture. METHODS: Chondrocytes isolated by enzymatic digestion of cartilage obtained from subjects undergoing knee replacement for OA (n = 6) or from normal ankle joints of tissue donors (n = 7) were cultured in alginate beads in serum-free medium and treated for 21 days with 100 ng/ml IGF-1, 100 ng/ml OP-1, or both. Controls were treated with vehicle alone. The cultures were evaluated for cell survival, cell number by DNA analysis, matrix production by particle exclusion assay, and level of accumulated proteoglycan by dimethylmethylene blue assay. RESULTS: After 21 days in serum-free alginate culture, survival of cells from OA cartilage was 65 +/- 2% (mean +/- SEM), while survival of cells from normal cartilage was significantly greater (82 +/- 3%). Treatment with either IGF-1 or OP-1 alone minimally improved survival, while the combination IGF +OP significantly improved survival, to 87 +/- 2% for OA cells and 95+/-1% for normal cells. Cell proliferation was noted only in the IGF+OP group; this was significant for both normal and OA cells ( approximately 2-fold increase in DNA levels). Matrix production, assessed by particle exclusion and by proteoglycan accumulation, was greatest in the cells treated with IGF + OP in both normal and OA cultures. When proteoglycan levels were corrected for cell numbers (mg proteoglycan/ng DNA), a significant increase over control was noted with OP-1 alone and IGF IGF-1 alone, in both normal and OA cultures, with the greatest levels in the combination group (3-fold increase over control). CONCLUSION: OP-1 was more potent than IGF-1 in stimulating proteoglycan production in both normal and OA cells. However, the best results were obtained with the combination, suggesting that combined therapy with IGF-1 and OP-1 may be an effective strategy for treating OA cartilage damage. PMID- 12905473 TI - High binding capacity of cyclophilin B to chondrocyte heparan sulfate proteoglycans and its release from the cell surface by matrix metalloproteinases: possible role as a proinflammatory mediator in arthritis. AB - OBJECTIVE: To study cyclophilin B, a protein newly identified as a secretion product of cultured chondrocytes, in the context of chondrocyte pathobiology. METHODS: Cyclophilin B was purified by sequential chromatographic processing of the secretion medium of cultured guinea pig chondrocytes. Its presence both at the surface of chondrocyte monolayers and in cartilage was demonstrated by immunohistochemistry. Binding sites at the surface of chondrocytes were characterized by Scatchard plot analysis using (125)I-labeled cyclophilin B, and by glycosidase treatments. The release of cyclophilin B from chondrocytes by activated matrix metalloproteinases (MMPs) was studied by Western blot analysis. RESULTS: Cyclophilin B was present at the surface of cultured chondrocytes and within cartilage, both in cells and in the extracellular matrix, with a particularly intense staining in the superficial layer. It was secreted constitutively by chondrocytes and cartilage explants. Its secretion was enhanced after treatment with its pharmacologic binding partner, cyclosporin A (CSA). Experiments with (125)I-labeled cyclophilin B demonstrated the presence of high capacity, low-affinity, NaCl-sensitive binding sites at the surface of chondrocytes. Cell-bound cyclophilin B could be released by heparinase treatment, demonstrating binding to pericellular heparan sulfate proteoglycans (HSPGs). Chondroitinase or keratanase treatments had no effect. MMPs 1, 2, 3, 9, and 13 released intact cyclophilin B from the cell surface, probably by cleavage of HSPGs. This effect was reversed by the broad-spectrum MMP inhibitor, marimastat. CONCLUSION: Cyclophilin B is a secreted CSA-binding protein involved in inflammatory events. It can induce chemotaxis in human neutrophils and T lymphocytes. The finding that cyclophilin B is an intrinsic component of cartilage and that it can be released by MMPs suggests that it has a role in the pathogenesis of arthritic diseases, even more so since its signaling receptor is present within the inflamed joint both on T cells and in the rheumatoid synovium. PMID- 12905474 TI - Arthritis self-management education programs: a meta-analysis of the effect on pain and disability. AB - OBJECTIVE: Some reports suggest that education programs help arthritis patients better manage their symptoms and improve function. This review of the published literature was undertaken to assess the effect of such programs on pain and disability. METHODS: Medline and HealthSTAR were searched for the period 1964 1998. The references of each article were then hand-searched for further publications. Studies were included in the meta-analysis if the intervention contained a self-management education component, a concurrent control group was included, and pain and/or disability were assessed as end points. Two authors reviewed each study. The methodologic attributes and efficacy of the interventions were assessed using a standardized abstraction tool, and the magnitude of the results was converted to a common measure, the effect size. Summary effect sizes were calculated separately for pain and disability. RESULTS: The search strategy yielded 35 studies, of which 17 met inclusion criteria. The mean age of study participants was 61 years, and 69% were female. On average, 19% of patients did not complete followup (range 0-53%). The summary effect size was 0.12 for pain (95% confidence interval [95% CI] 0.00, 0.24) and 0.07 for disability (95% CI 0.00, 0.15). Funnel plots indicated no significant evidence of bias toward the publication of studies with findings that showed reductions in pain or disability. CONCLUSION: The summary effect sizes suggest that arthritis self-management education programs result in small reductions in pain and disability. PMID- 12905475 TI - Frequency of osteopenia in adolescents with early-onset juvenile idiopathic arthritis: a long-term outcome study of one hundred five patients. AB - OBJECTIVE: To determine the frequency of low bone mineral content (BMC) and low bone mineral density (BMD) as long-term complications in adolescents with early onset juvenile idiopathic arthritis (JIA), and to identify disease variables, patient characteristics, and biochemical bone markers related to low bone mass. METHODS: One hundred five (87%) of 121 adolescent patients with early-onset JIA (ages 13-19 years, 80 girls and 25 boys, mean age at onset of JIA 2.8 years), from a cohort first admitted to the hospital between 1980 and 1985, were assessed after a mean disease duration of 14.2 years. BMC and BMD of the total body, the lumbar spine at L2-L4, and the femoral neck were measured by dual-energy x-ray absorptiometry. Age- and sex-specific reference values from a pooled, healthy reference population were used to calculate Z scores. Low bone mass was defined as a Z score less than -1 SD. RESULTS: Among the 103 adolescent JIA patients who underwent total-body imaging, 41% had low total-body BMC and 34% had low total body BMD. Compared with adolescent JIA patients who had normal total-body BMC, those with low BMC had lower mean weight (P < 0.001), height (P < 0.001), lean mass (P < 0.001), and remission rates (P = 0.016), had longer duration of active disease (P = 0.013), had higher numbers of active and mobility-restricted joints (P < 0.001 and P = 0.001, respectively), had more disability (P = 0.011), had higher frequencies of joint erosions (P < 0.001), and had higher erythrocyte sedimentation rates (P = 0.033). In multiple linear regression analyses of total body BMC, 88% of the variance was explained by the duration of active disease, the number of joints with restricted mobility, the bone area, urinary deoxypyridinoline values, age, weight, and height. CONCLUSION: Forty-one percent of the adolescents with early-onset JIA had low bone mass >11 years after disease onset. The development of low total-body BMC was related to the duration of active disease, disease severity, measures of bone resorption, weight, and height. PMID- 12905476 TI - Long-term efficacy and safety of infliximab in the treatment of ankylosing spondylitis: an open, observational, extension study of a three-month, randomized, placebo-controlled trial. AB - OBJECTIVE: Treatment of ankylosing spondylitis (AS) with infliximab, an anti tumor necrosis factor alpha monoclonal antibody, was shown to be efficacious in patients with active disease during a 3-month treatment period. The purpose of this study was to evaluate the efficacy and safety of infliximab treatment of AS for a 1-year period. METHODS: This study was an open, observational, extension study of a 3-month, randomized, placebo-controlled trial. All patients who had tolerated infliximab (infliximab/infliximab group) or placebo (placebo/infliximab 12-week crossover group) therapy for 3 months entered the open extension trial (n = 65). Infliximab was administered at a dosage of 5 mg/kg every 6 weeks after the induction phase (weeks 0, 2, and 6). The primary end point was a 50% improvement in the Bath Ankylosing Spondylitis Disease Activity Index (BASDAI). RESULTS: At week 54, a total of 54 of the 69 patients (78%) continued to take infliximab. The intent-to-treat primary efficacy analysis at week 54 showed that 47% of patients in the infliximab/infliximab group (95% confidence interval 31-63) and 51% of the patients in the placebo/infliximab group (95% confidence interval 36-67) achieved 50% improvement in BASDAI scores. In the analysis of those who completed the study, the mean BASDAI scores improved between weeks 0 and 54 in both treatment groups: from 6.6 to 2.4 in the infliximab/infliximab group and from 6.3 to 2.6 in the placebo/infliximab group. The dosage of nonsteroidal antiinflammatory drugs was reduced in approximately 70% of the patients. There were significant improvements in measures of functioning, metrologic parameters, and quality of life. Between weeks 12 and 54, a total of 4 patients had serious adverse events that were possibly related to infliximab and resulted in their discontinuing the study. CONCLUSION: Infliximab therapy in AS patients resulted in a rapid and significant improvement in BASDAI scores (>50% improvement) and a durable response for 1 year. The safety profile of infliximab in AS was comparable to that observed in the postmarketing experience for the approved indications. PMID- 12905477 TI - Identification of major loci controlling clinical manifestations of ankylosing spondylitis. AB - OBJECTIVE: To identify genomic regions linked with determinants of age at symptom onset, disease activity, and functional impairment in ankylosing spondylitis (AS). METHODS: A whole genome linkage scan was performed in 188 affected sibling pair families with 454 affected individuals. Traits assessed were age at symptom onset, disease activity assessed by the Bath Ankylosing Spondylitis Disease Activity Index (BASDAI), and functional impairment assessed by the Bath Ankylosing Spondylitis Functional Index (BASFI). Parametric and nonparametric quantitative linkage analysis was performed using parameters defined in a previous segregation study. RESULTS: Heritabilities of the traits studied in this data set were as follows: BASDAI 0.49 (P = 0.0001, 95% confidence interval [95% CI] 0.23-0.75), BASFI 0.76 (P = 10(-7), 95% CI 0.49-1.0), and age at symptom onset 0.33 (P = 0.005, 95% CI 0.04-0.62). No linkage was observed between the major histocompatibility complex (MHC) and any of the traits studied (logarithm of odds [LOD] score <1.0). "Significant" linkage (LOD score 4.0) was observed between a region on chromosome 18p and the BASDAI. Age at symptom onset showed "suggestive" linkage to chromosome 11p (LOD score 3.3). Maximum linkage with the BASFI was seen at chromosome 2q (LOD score 2.9). CONCLUSION: In contrast to the genetic determinants of susceptibility to AS, clinical manifestations of the disease measured by the BASDAI, BASFI, and age at symptom onset are largely determined by a small number of genes not encoded within the MHC. PMID- 12905478 TI - In primary Sjogren's syndrome, HLA class II is associated exclusively with autoantibody production and spreading of the autoimmune response. AB - OBJECTIVE: To reevaluate, in a large series of patients with Sjogren's syndrome (SS) recruited from 2 French centers, the question of whether HLA is associated with SS itself or with a pattern of secretion of autoantibodies. METHODS: One hundred forty-nine white patients fulfilling the American-European Consensus Group criteria for SS were divided into 3 subgroups, according to their anti Ro/SSA and anti-La/SSB status, as follows: group 1 (n = 53), no antibody; group 2 (n = 46), anti-SSA only; group 3 (n = 50), both anti-SSA and anti-SSB. Patients were compared with 222 unrelated healthy subjects representative of the white population in France. RESULTS: Comparisons between the 149 SS patients and 222 controls confirmed the association of SS with DRB1*03 (the frequency was 25% in patients versus 10% in controls) and DQB1*02 (32% versus 22%). The association between HLA and SS was restricted to patients with anti-SSA and/or anti-SSB; no association with HLA was observed in patients in group 1 (no antibody). The frequency of HLA-DRB1*15 was highest in group 2 (24%), compared with 11% in group 1 and 11% in controls, whereas the frequency of HLA-DRB1*03 was highest in group 3 (44%), compared with 12% in group 1, 19% in group 2, and 10% in controls. Group 2 and group 3 had more clinical and biologic markers of activity than did group 1 but were not clinically different. HLA alleles were not associated with clinical features of the disease, and were associated with only some biologic features: rheumatoid factor positivity, increased serum IgG, and thrombocytopenia were associated with HLA-DRB1*03, and neutropenia was associated with DQB1*01. CONCLUSION: HLA class II markers confer genetic susceptibility to Sjogren's syndrome. The association between HLA and SS is restricted to patients with anti SSA and/or anti-SSB antibodies; HLA is not associated with SS in patients without these autoantibodies. The absence of a difference in disease severity between groups 2 and 3, as well as the restricted association of HLA-DRB1*03 in group 3, strongly suggest that HLA alleles predispose to autoantibody secretion, without being associated with clinical outcome. HLA class II phenotype might support epitope spreading: HLA-DR15 favors anti-SSA synthesis, whereas HLA-DR3 is associated with both anti-SSA and anti-SSB production. PMID- 12905479 TI - Prevalence, incidence, survival, and disease characteristics of systemic sclerosis in a large US population. AB - OBJECTIVE: To estimate the prevalence, incidence, survival, and disease characteristics of systemic sclerosis (SSc) in the Detroit tricounty area. METHODS: A census of SSc cases for the period 1989-1991 was conducted in the Detroit area, using multiple sources for case identification. Diagnoses were verified by medical record review. Capture-recapture analysis was used to estimate the total SSc population. Cases of localized scleroderma (morphea and linear disease) were excluded. RESULTS: Based on 706 verified cases of SSc, prevalence was initially estimated to be 242.0 cases per million adults (95% confidence interval [95% CI] 213-274), with an annual incidence of 19.3 new cases per million adults per year (95% CI 12.4-30.2). Capture-recapture analysis, based on the degree of overlap of verified cases among multiple sources, resulted in a revised prevalence estimate of 276 cases per million adults (95% CI 245-310). Sex and race-specific prevalence estimates were significantly higher for women than for men, and for blacks than for whites. The average age at diagnosis was significantly younger for blacks than for whites. Compared with white patients, black patients were almost twice as likely to have diffuse disease (prevalence proportion ratio 1.86, 95% CI 1.48-2.35). Median survival was approximately 11 years. Factors negatively affecting survival included male sex (hazard ratio 1.81, 95% CI 1.29-2.55) and older age at diagnosis (hazard ratio 1.04, 95% CI 1.03-1.05). CONCLUSION: This study establishes baseline estimates of SSc occurrence and characteristics in a large US cohort consisting primarily of black adults and white adults. These data should facilitate research regarding the role of geographic, ethnic, racial, and environmental factors for this disease in comparison populations. PMID- 12905480 TI - Effects of oral cyclophosphamide and prednisolone therapy on the endothelial functions and clinical findings in patients with early diffuse systemic sclerosis. AB - OBJECTIVE: The endothelial damage of microvascular structures in systemic sclerosis (SSc; scleroderma) is associated with increased levels of endothelial adhesion molecules and endothelium-associated cytokines, including E-selectin and thrombomodulin. Although there is still no ideal specific pharmacologic therapy for SSc, cyclophosphamide has resulted in clinical improvement in patients with SSc-related active alveolitis. This study was designed to assess the expression of E-selectin and thrombomodulin in patients with early diffuse SSc, and to investigate the effects of oral cyclophosphamide combined with prednisolone therapy on the levels of these endothelium-associated cytokines and on the patients' clinical outcomes. METHODS: Thirteen patients with early diffuse SSc were treated with oral cyclophosphamide (2-2.5 mg/kg/day) and methylprednisolone (30 mg/every other day) for 1 year. The outcomes were determined as clinical (skin score) and laboratory parameters (including the erythrocyte sedimentation rate, complete blood cell count, levels of C-reactive protein, antinuclear antibody, anti-double-stranded DNA, rate of creatinine clearance, and findings on pulmonary function tests, esophageal manometry, and echocardiography). The concentrations of E-selectin and thrombomodulin were measured in the pretreatment and posttreatment serum samples from the SSc patients and from 12 healthy adults as controls. RESULTS: In the patients with early diffuse SSc, pretreatment and posttreatment mean levels of E-selectin were 51 ng/ml (range 34.2-135.5) and 33.4 ng/ml (range 23-62.5), respectively (P = 0.01), and those of thrombomodulin were 82 ng/ml (range 35.8-120.5) and 74.6 ng/ml (range 23.3-91.3), respectively (P = 0.016). Clinical and laboratory parameters (the skin score and measures of pulmonary function [forced vital capacity and diffusing capacity for carbon monoxide]) were also improved (P < 0.05 for each) at the end of the followup period. CONCLUSION: Combination therapy with cylophosphamide plus prednisolone is effective in the treatment of early diffuse SSc. Circulating levels of E-selectin and thrombomodulin not only demonstrate the extent of endothelial injury and/or activation, but also could be a useful marker to monitor the disease activity in SSc. PMID- 12905481 TI - Occurrence of an activated, profibrotic pattern of gene expression in lung CD8+ T cells from scleroderma patients. AB - OBJECTIVE: Pulmonary fibrosis is a major cause of death in scleroderma patients. Previous studies have shown an increase in CD8+ T cells in the lungs of scleroderma patients. In the present study, we sought to determine whether activated CD8+ T cells contribute to pulmonary fibrosis in scleroderma patients through the production and activation of profibrotic mediators. METHODS: CD8+ cells were isolated from bronchoalveolar lavage fluid obtained from 19 scleroderma patients and 7 healthy subjects. The phenotype of these cells was determined using DNA array technology. Expression of selected genes was confirmed in real-time polymerase chain reaction and enzyme-linked immunosorbent assay experiments. RESULTS: Hierarchical clustering of gene expression profiles revealed 2 groups of subjects. Group 1 consisted of 11 patients (8 with and 3 without lung inflammation). Group 2 consisted of 15 subjects (7 healthy controls and 2 patients with and 6 without lung inflammation). Gene expression in group 1 indicated T cell activation, a type 2 phenotype, production of profibrotic factors and matrix metalloproteinases, and reduced activation-induced cell death. Increased expression of beta6 integrin messenger RNA by CD8+ T cells in group 1 suggested the possibility that these T cells might induce cell-contact-dependent activation of latent transforming growth factor beta (TGFbeta). CONCLUSION: A subset of scleroderma patients at higher risk of progressive lung disease have activated, long-lived CD8+ T cells in their lungs that could promote fibrosis directly, through production of profibrotic factors such as interleukin-4 and oncostatin M, as well as indirectly, through activation of TGFbeta. PMID- 12905482 TI - Molecular mechanisms of interleukin-4-induced up-regulation of type I collagen gene expression in murine fibroblasts. AB - OBJECTIVE: There is evidence that interleukin-4 (IL-4) plays a major role in the induction of extracellular matrix protein synthesis in fibrotic disease. We therefore examined the effect of IL-4 on collagen synthesis in primary fibroblasts isolated from normal and TSK/+ mice, which spontaneously develop a scleroderma-like syndrome characterized by diffuse cutaneous hyperplasia. METHODS: Expression of the IL-4 receptor was determined by flow cytometry and Western blotting. The IL-4 signal transduction cascade was analyzed by Western blotting. We assessed the role of signal transducer and activator of transcription 6 (STAT-6) in IL-4 induction of alpha2(I) collagen promoter activity and message levels via luciferase reporter assay and real-time polymerase chain reaction. The activation status of the transcription factors activator protein 1 (AP-1) and Sp-1 upon stimulation with IL-4 in normal and TSK/+ fibroblasts was examined by electrophoretic mobility shift assay. RESULTS: Flow cytometry and Western blotting showed that IL-4 receptor alpha expression was elevated in TSK/+ fibroblasts compared with normal fibroblasts. After IL-4 stimulation, janus-activated kinase 1 (JAK-1) and JAK-2 were phosphorylated to a greater degree in TSK/+ fibroblasts than in C57BL/6 fibroblasts. TSK/+ fibroblasts appeared to be hyperresponsive to IL-4, displaying increased synthesis of alpha1(I) collagen messenger RNA (mRNA), collagen protein, and activity of a luciferase reporter construct containing the -300 to +54 murine alpha2(I) collagen promoter. Overexpression of STAT-6 enhanced this effect, whereas expression of a dominant-negative STAT-6 abrogated the ability of IL-4 to induce alpha1(I) collagen mRNA in TSK/+ fibroblasts. Moreover, IL-4 induced increased DNA binding activity of transcription factors that are important for collagen synthesis. CONCLUSION: Our observations indicate that IL-4 has a profound effect on several factors that have been identified as playing major roles in the regulation of collagen synthesis and suggest that IL-4 increases the expression of type I collagen through a mechanism involving the activation of transcription factors that bind to and activate collagen promoter. PMID- 12905483 TI - Global surface ultraviolet radiation intensity may modulate the clinical and immunologic expression of autoimmune muscle disease. AB - OBJECTIVE: To determine if geoclimatic factors may influence the nature and frequency of dermatomyositis (DM), polymyositis, and associated autoantibodies around the world. METHODS: We assessed, in the first global evaluation of these conditions, the relationship between 13 geoclimatic variables that may modulate disease and the relative proportion of DM and its associated autoantibody anti-Mi 2, directed against an SNF2-superfamily helicase associated with the nucleosome remodeling and histone acetylation and deacetylation complex, in a global myositis population. Altogether, 919 consecutive patients from populations at 15 locations were studied. RESULTS: Univariate and multivariate analyses demonstrated that of the variables evaluated, surface ultraviolet (UV) radiation intensity (irradiance) most strongly contributed to the relative proportion of DM and was strongly related to the proportion of anti-Mi-2 autoantibodies (weighted r = 0.939, P < 4 x 10(-7) and weighted r = 0.69, P = 0.02, respectively). Published ethnogeographic immunogenetic allele frequencies imply that the striking differences in the proportion of DM- and DM-specific autoantibodies observed around the world are not the result of inherent global variations in known genetic risk factors. CONCLUSION: These data suggest that UV radiation exposure may modulate the clinical and immunologic expression of an autoimmune disease in different populations around the world. PMID- 12905484 TI - Bone mineral density in juvenile dermatomyositis: assessment using dual x-ray absorptiometry. AB - OBJECTIVE: To determine the bone mineral density (BMD) status of our juvenile dermatomyositis (DM) population and to compare the frequency of osteopenia in patients with active disease requiring corticosteroids with that in patients with inactive disease who are not receiving corticosteroids. METHODS: Medical charts of all children diagnosed as having juvenile DM at our institution between 1989 and 1999 were reviewed for demographic and clinical data, including disease activity and duration of corticosteroid therapy. BMD measurements of the lumbar spine (L1-L4) were performed using dual x-ray absorptiometry (DXA). Z scores were calculated from the BMD data for comparison with published normative data. RESULTS: A total of 15 patients were assessed: 10 with active disease, and 5 with inactive disease who had not taken corticosteroids for an average of 6.0 years (range 3.4-8.1 years). Baseline BMD measurements demonstrated osteopenia or frank osteoporosis in the majority of patients, including 6 of the 10 patients with active disease and 4 of the 5 patients whose disease was in remission. Fourteen patients had serial BMD measurements. Persistent or worsening osteopenia was documented in all patients who had ongoing active disease, except for 3 patients who had been treated with bisphosphonates because of vertebral compression fractures. CONCLUSION: Osteopenia is common in patients with juvenile DM, and it usually worsens with ongoing disease. It can persist for many years after the disease enters remission. Bisphosphonates appeared to beneficially affect bone mineralization in our patients. Treatment to prevent the long-term complications of osteoporosis in patients with juvenile DM should be considered and requires further study. PMID- 12905485 TI - Limited versus severe Wegener's granulomatosis: baseline data on patients in the Wegener's granulomatosis etanercept trial. AB - OBJECTIVE: To report baseline data on 180 patients with Wegener's granulomatosis (WG) enrolled in the WG Etanercept Trial (WGET), and to examine demographic and clinical differences between patients with limited disease and those with severe disease. METHODS: Definitions of limited and severe disease were agreed upon by consensus of the investigators at a pretrial meeting and were incorporated into the protocol as a stratification criterion. These data were applied prospectively to the WGET patient cohort, based on clinical features and the intention to treat patients according to disease activity. Data related to disease onset, date of diagnosis, clinical features, antineutrophil cytoplasmic antibody assays, tissue biopsy findings, and other medical history were collected on a baseline medical history form. Physician-investigators from each center participated in the development of this form, and all were certified in its use prior to the start of the trial. Selected data on patients who were screened for the trial but were not enrolled were also collected. RESULTS: Several significant differences between the limited and severe disease subsets were observed. Patients with limited disease were nearly a decade younger at disease onset compared with patients with severe disease. Thirty-three percent of patients with severe disease were women, compared with 58% of those with limited disease. Despite their younger age at symptom onset, patients with limited disease tended to have longer disease duration, a greater likelihood of experiencing exacerbation of previous disease following a period of remission, and a higher prevalence of destructive upper respiratory tract disorders at the time of enrollment (e.g., saddle-nose deformity). Patients with limited WG were less likely than those with severe disease to have antibodies to either proteinase 3 or myeloperoxidase. Patients with severe disease had a higher likelihood of previous thyroid disease, particularly either Graves' disease or Hashimoto thyroiditis, suggesting the possibility of different pathogenetic factors within these disease subsets. Other observed differences between these subsets, such as the greater frequency of alveolar hemorrhage in the severe disease group, were related to the a priori definitions of limited and severe disease. CONCLUSION: There are significant differences between patients with limited WG and those with severe WG with regard to sex, age, the likelihood of recurrent disease, the risk of damage in certain organ systems, and, possibly, etiologic factors. These differences (and perhaps other differences that are currently unrecognized) in patient subsets may have implications for mechanisms of pathogenesis, prognosis, response to treatment, and the design of future clinical investigations. PMID- 12905486 TI - Role of S100A8 and S100A9 in neutrophil recruitment in response to monosodium urate monohydrate crystals in the air-pouch model of acute gouty arthritis. AB - OBJECTIVE: To examine the role of chemokines, S100A8, and S100A9 in neutrophil accumulation induced by the causative agent of gout, monosodium urate monohydrate (MSU) crystals. METHODS: MSU crystal-induced neutrophil migration was studied in the murine air-pouch model. Release of chemokines, S100A8, S100A9, and S100A8/A9 in response to MSU crystals was quantified by enzyme-linked immunosorbent assays. Recruited cells were counted following acetic blue staining, and the subpopulations were characterized by Wright-Giemsa staining of cytospins. RESULTS: MSU crystals induced the accumulation of neutrophils following injection in the air pouch, which correlated with the release of the chemokines CXCL1, CXCL2, CCL2, and CCL3. However, none of these was found to play an important role in neutrophil migration induced by MSU crystals by passive immunization with antibodies directed against each chemokine. S100A8, S100A9, and S100A8/A9 were also found at high levels in the pouch exudates following injection of MSU crystals. In addition, injection of S100A8, S100A9, or S100A8/A9 led to the accumulation of neutrophils in the murine air pouch, demonstrating their proinflammatory activities in vivo. Passive immunization with anti-S100A8 and anti-S100A9 led to a total inhibition of the accumulation of neutrophils. Finally, S100A8/A9 was found at high concentrations in the synovial fluid of patients with gout. CONCLUSION: S100A8 and S100A8/A9 are essential to neutrophil migration induced by MSU crystals. These results suggest that they might be involved in the pathogenesis of gout. PMID- 12905487 TI - Bisphosphonate resistance in Paget's disease of bone. AB - OBJECTIVE: To determine whether resistance to one bisphosphonate predicts resistance to another bisphosphonate. METHODS: One hundred patients with Paget's disease were treated with intravenous (IV) pamidronate. The initial dose was 120 mg, followed by further doses of 240 mg, until either biochemical remission was achieved or a total dose of 1 gm was given. Biochemical remission was defined as an alkaline phosphatase level within the reference range. Patients whose disease failed to respond to pamidronate were then treated with alendronate for 6 months. Patients whose disease failed to respond to alendronate were given either tiludronate for 3 months, or clodronate for 6 months. RESULTS: Sixteen of the 100 patients treated with pamidronate failed to achieve a biochemical response despite a cumulative dose of 1 gm. Of the 16 nonresponders, 1 died of an unrelated cause, and the remaining 15 patients were treated with alendronate. In 2 of these patients, the treatment was changed to another bisphosphonate because of gastrointestinal intolerance to alendronate. Of the remaining 13 patients, 9 (69%) achieved full biochemical remission. In 4 other patients, both pamidronate and alendronate therapy were unsuccessful (1 patient responded to tiludronate, tiludronate therapy was unsuccessful in 1, clodronate was unsuccessful in 1, and 1 patient elected to receive no further treatment). Of the 2 patients who could not receive alendronate because of gastrointestinal intolerance, 1 achieved normalization with tiludronate, and a repeat course of pamidronate was unsuccessful in the other. In total, 73% of patients in whom initial treatment with IV pamidronate was unsuccessful responded to a change in bisphosphonate treatment. CONCLUSION: Failure to achieve biochemical normalization is likely to be specific to the individual drug rather than indicative of bisphosphonate class resistance. PMID- 12905488 TI - Familial Mediterranean fever among patients from Karabakh and the diagnostic value of MEFV gene analysis in all classically affected populations. AB - OBJECTIVE: Familial Mediterranean fever (FMF) is an autosomal-recessive disorder that is common in Armenian, Turkish, Arab, and Sephardic Jewish populations. Its clinical diagnosis is one of exclusion, with the patients displaying nonspecific symptoms related to serosal inflammation. MEFV gene analysis has provided the first objective diagnostic criterion for FMF. However, in the absence of an identified mutation (NI/NI genotype), both the sensitivity of the molecular analyses and the involvement of the MEFV gene in FMF are called into question. The present study was designed to further evaluate the diagnostic value of MEFV analysis in another population of Mediterranean extraction. METHODS: The MEFV gene was screened for mutations in 50 patients living in Karabakh (near Armenia) who fulfilled the established criteria for FMF. In addition, we analyzed published series of patients from the above-mentioned at-risk populations. RESULTS: The mutation spectrum in Karabakhian patients, which consisted of only 6 mutations (with 26% of NI alleles), differed from that reported in Armenian patients. Strikingly, among patients from Karabakh and among all classically affected populations, the distribution of genotypes differed dramatically from Hardy-Weinberg equilibrium (P = 0.0016 and P < 0.00001, respectively). These results, combined with other population genetics-based data, revealed the existence of an FMF-like condition that, depending on the patients' ancestry, was shown to affect 85-99% of those with the NI/NI genotype. CONCLUSION: These data illuminate the meaning of negative results of MEFV analyses and show that in all populations evaluated, most patients with the NI/NI genotype had disease that mimicked FMF and was unrelated to the MEFV gene. Our findings also demonstrate the high sensitivity of a search for very few mutations in order to perform a molecular diagnosis of MEFV-related FMF. PMID- 12905489 TI - A comparative genetic analysis between collagen-induced arthritis and pristane induced arthritis. AB - OBJECTIVE: To compare the genetic regulation of collagen-induced arthritis (CIA) with that of pristane-induced arthritis (PIA) in rats. METHODS: A genome-wide linkage analysis of an (E3 x DA)DA backcross of rats with CIA (n = 364 male rats; the same strain combinations as previously used to determine the genetic control of PIA) was performed. The strongest loci in both CIA and PIA (i.e., Cia12/Pia4 and Cia13/Pia7) were isolated in congenic strains. Susceptibility in both congenic strains was tested in rats with CIA and in rats with PIA. RESULTS: We found a striking, although not complete, similarity of the arthritis-controlling loci in CIA and in PIA, as well as the previously defined loci associated with cartilage destruction, antibody production, and the acute-phase response. All major PIA quantitative trait loci (QTLs) identified in early severe arthritis were also strong regulators of CIA. The 2 strongest QTLs, Cia12/Pia4 on chromosome 12 and Cia13/Pia7 on chromosome 4, were also analyzed in congenic strains with DA or E3 as the background genome. Consistent with the results of linkage analysis, the congenic strain experiments showed that the chromosome 4 locus was more penetrant in CIA than in PIA, while the chromosome 12 locus almost completely dominated the control of PIA severity. CONCLUSION: The underlying genetic control of CIA was found to have many, but not all, pathogenic mechanisms in common with PIA, despite the use of a cartilage-specific antigen (type II collagen) to induce CIA but not PIA. PMID- 12905490 TI - Use of a novel elution regimen reveals the dominance of polyreactive antinuclear autoantibodies in lupus kidneys. AB - OBJECTIVE: The autoantibody specificities that dominate the deposits in lupus kidneys remain unclear. Reasoning that previously utilized elution buffers such as acidic glycine and ammonium thiocyanate may not have been maximally effective in eluting all Ig deposits from the kidneys, this study was conducted to experiment with a stronger dissociating agent, urea-glycine. METHODS: Seven antinuclear antibody-positive, nephritic female (SWR x NZB)F(1) (SNF1) lupus mice were selected for the elution study. Deposited Ig was eluted from their kidneys using 3 different elution buffers: 0.15M glycine-HCl buffer, 1.3M ammonia thiocyanate/0.15M glycine-HCl buffer, and 5M urea/0.15M glycine-HCl buffer. All eluates were tested for specificity against a variety of nuclear and glomerular antigens. RESULTS: Compared with conventional elution buffers, the urea-based regimen eluted severalfold more IgG and IgM antinuclear antibodies from the kidneys of nephritic SNF1 lupus mice. IgG anti-double-stranded DNA (anti-dsDNA) antibodies were not only the most prevalent species in these renal deposits, they were also heavily enriched in the kidneys, relative to the corresponding serum levels. A substantial fraction of the anti-single-stranded DNA and antihistone/DNA (but not antihistone) reactivity in these eluates was due to cross-reactive anti-dsDNA antibodies. No reactivity with SSA, SSB, Sm, RNP, Jo-1, Scl-70, or ribosomal P antigens could be demonstrated in these eluates. Importantly, the urea-glycine eluates differed from the conventional eluates in having significantly greater reactivity to glomerular substrate and laminin. CONCLUSION: This novel urea-based elution provides further support for the dominance of antibodies in lupus kidneys, with strong polyreactivity to DNA and glomerular substrate. PMID- 12905491 TI - Defects in the regulation of B cell apoptosis are required for the production of citrullinated peptide autoantibodies in mice. AB - OBJECTIVE: Protein deimination, a process to modify arginine residues to citrulline by the addition of a neutral oxygen group, is associated with apoptosis. The presence of autoantibodies recognizing citrullinated peptides is highly specific to rheumatoid arthritis (RA) and is therefore a useful marker for the early diagnosis of RA. In this study, we explored whether anti-cyclic citrullinated peptide (anti-CCP) autoantibodies are produced in several experimental models of autoimmune diseases in mice. METHODS: The levels of anti CCP autoantibodies were analyzed by enzyme-linked immunosorbent assay in several lupus-prone strains of mice, in animals with type II collagen (CII)-induced arthritis, and after induction of neonatal tolerance to alloantigens. RESULTS: We observed the production of these autoantibodies in 2 different lupus-prone mice, MRL-lpr/lpr and (NZW x B6)F(1)-hbcl-2 transgenic mice, characterized by the presence of abnormalities in the regulation of B cell apoptosis. Other genetic defects, determining autoimmune susceptibility, present in MRL and NZW mice were additionally required for anti-CCP autoantibody production. The induction of autoantibodies in normal BALB/c mice injected at birth with semiallogeneic spleen cells from (BALB/c x B6)F(1)-hbcl-2 transgenic mice suggested that these additional autoimmune defects may be related, at least in part, to the establishment of abnormal interactions between T cells and B cells. In addition, anti-CCP autoantibodies were not produced in the course of CII-induced arthritis, an experimental model of RA in mice. CONCLUSION: Our study provides evidence for the association between defects in the regulatory cell death machinery of B lymphocytes and the production of certain autoantibody specificities. PMID- 12905492 TI - Dendritic cell uptake of human apoptotic and necrotic neutrophils inhibits CD40, CD80, and CD86 expression and reduces allogeneic T cell responses: relevance to systemic vasculitis. AB - OBJECTIVE: There is a breakdown of tolerance to neutrophil components during systemic vasculitis, which is marked by autoantibodies and T cells with specificity for proteinase 3 or myeloperoxidase, expressed on the surface of apoptotic neutrophils. This study was undertaken to investigate the effects of human apoptotic and necrotic neutrophils on human dendritic cell (DC) phenotype and ability to stimulate allogeneic T cell proliferation. METHODS: DCs were generated from human peripheral blood mononuclear cells and allowed to interact with human apoptotic and necrotic neutrophils in the presence or absence of tumor necrosis factor alpha (TNFalpha). Effects on DC phenotype and ability to stimulate T cell proliferation were observed. RESULTS: Immature DCs engulfed apoptotic and necrotic neutrophils, resulting in up-regulation of CD83 and class II major histocompatibility complex molecules, but down-regulation of CD40, CD80, and CD86, and a decreased ability to stimulate T cell proliferation. When TNFalpha was added in combination with apoptotic neutrophils, the inhibitory effects were overcome to some extent. CONCLUSION: Our results suggest that DC uptake of apoptotic or necrotic neutrophils alone does not shift the immune response from tolerance to autoimmunity in systemic vasculitis. However, cytokines found at sites of inflammation in vasculitis patients may act as maturation factors for DCs, and in combination with apoptotic neutrophils, may lead to an autoimmune phenotype. PMID- 12905494 TI - Tumor necrosis factor receptor-associated periodic syndrome characterized by a mutation affecting the cleavage site of the receptor: implications for pathogenesis. PMID- 12905495 TI - Availability of drugs should be disclosed by manufacturers. PMID- 12905496 TI - Lymphomas after tumor necrosis factor antagonist therapy: comment on the article by Brown et al. PMID- 12905493 TI - Cytokines elicited by T cell epitopes from a synovial autoantigen: altered peptide ligands can reduce interferon-gamma and interleukin-10 production. AB - OBJECTIVE: To explore the cytokine responses associated with T cell epitopes from human cartilage glycoprotein 39 (HC gp-39) and the potential for modifying cytokine secretion using altered peptide ligands (APLs). METHODS: Draining lymph node cells were harvested from HLA-DR*0401 transgenic mice that had been immunized with HC gp-39. Cytokine responses to 5 previously identified HLA DR*0401-restricted HC gp-39 T cell epitopes were studied in vitro. The anchor and T cell receptor (TCR) contact residues of peptide 322-337 were identified, and this information was used to design alanine-substituted APLs. T cells were primed in vivo with wild-type peptide 322-337, restimulated with wild-type peptide or APLs, and the cytokine profiles were compared. RESULTS: Restimulation with individual peptides elicited distinct cytokine profiles. HC gp-39 (peptide 322 337) elicited a dominant interferon-gamma (IFNgamma) response. Residues within the core (positions P1-P9) 322-337 peptide sequence were critical for T cell recognition. Surprisingly, the N-terminal flanking region was also important for recognition by 6 of 10 specific T cell hybridomas. Substitutions of charged TCR contact residues in the 322-337 core epitope (E332A and K335A) were associated with a significant reduction in the IFNgamma and interleukin-10 (IL-10) stimulation indices. Restimulation with peptides W325A and V326A was also associated with a trend toward reduced IFNgamma and IL-10 secretion. In contrast, restimulation with peptide D330N elicited cytokine profiles more comparable with those resulting from restimulation with wild-type peptide. CONCLUSION: This study indicates that APLs of a proinflammatory HC gp-39 T cell epitope may be used to alter the cytokine response from a memory T cell population. PMID- 12905497 TI - Expression of the interferon-gamma-inducible 10-kd protein and CXC receptor 3 in the salivary gland lesions of patients with Sjogren's syndrome: comment on the article by Ogawa et al. PMID- 12905500 TI - Citations and reference lists: for whom and for what purpose? PMID- 12905501 TI - Improved antenatal detection of hemorrhagic/ischemic intracranial lesions using transvaginal ultrasonography and fetal magnetic resonance imaging. PMID- 12905502 TI - Peri- and intraventricular cerebral sonography in second- and third-trimester high-risk fetuses: a comparison with neonatal ultrasound and relation to neurological development. AB - OBJECTIVES: To determine whether periventricular leukomalacia (PVL) and peri- and intraventricular hemorrhages can be detected in fetuses at risk for preterm birth and to establish the clinical significance of this finding. METHODS: Prenatal cerebral sonography was performed in 26 fetuses at risk for uteroplacental insufficiency on the day of inclusion into the study and weekly until delivery. Neonatal cerebral ultrasound examination was performed within 24 h of birth and biweekly until discharge. The infants underwent standardized neurological examinations. RESULTS: During 30 observations, 21/26 fetuses presented in a cephalic position. Successful visualization in the coronal and sagittal planes was significantly correlated with that of a control population (r = 0.615; 95% CI, 0.390-0.771). In 20/21 fetuses at least one observation was adequate for analysis. Eleven cerebral abnormalities were found in nine fetuses, periventricular echodensities in four, intraventricular hemorrhage in two, and localized thalamic densities in five. In six of these infants ultrasound abnormalities persisted after birth. Neurological follow-up at 24 months demonstrated abnormalities in three infants, two infants were normal and one was lost to follow-up. The three infants with normal ultrasound results after birth had a normal follow-up. Five infants with normal antenatal ultrasound results showed ultrasound abnormalities during the neonatal period; four of them developed PVL Grade 1 and one developed PVL Grade 2. The follow-up of two of these infants was normal, two died and one developed spastic tetraplegia. Abnormal antenatal brain sonography was significantly correlated with gestational age at birth (r = 0.487; P = 0.029). CONCLUSION: Peri- and intraventricular echodensities can be detected reliably before birth in fetuses at high risk for uteroplacental insufficiency. When abnormalities persist, there is a high risk for an adverse outcome. PMID- 12905503 TI - Outcome of antenatally diagnosed intracranial hemorrhage: case series and review of the literature. AB - OBJECTIVES: Prenatal diagnosis of intracranial hemorrhage (ICH) has been widely reported. Hemorrhages may occur either within the cerebral ventricles, subdural space or infratentorial fossa. The aim of this study was to determine the sonographic criteria for the diagnosis of fetal ICH, the role of in utero magnetic resonance imaging (MRI) and the outcome of this condition. METHODS: The archives of our ultrasound laboratory and the literature were searched for all cases of antenatally diagnosed ICH. A grading system was used to classify the intraventricular lesions as suggested in postnatal sonographic studies. RESULTS: Adding our series of 16 fetuses to the 93 cases identified in the literature, a group of 109 fetal ICHs was obtained: 89 were intracerebral (79 intraventricular and 10 infratentorial) and 20 were subdural hemorrhages. Intraventricular lesions were mostly classified as severe (32 each for Grades III and IV). In 27 cases antenatal MRI was performed additionally to ultrasound and confirmed the sonographic findings. Of the entire group, 65 infants (59%) were reported to be alive 1 month after birth (51 intraventricular hemorrhages, three infratentorial hemorrhages, 11 subdural hematomas). At 12 months, of the 48 infants whose follow up was available, 25 or 52% were judged neurologically normal (17/36 or 47% among the intraventricular hemorrhages, 6/9 or 66% among the hematomas, and 2/3 or 66% among the infratentorial hemorrhages). CONCLUSIONS: Fetal ICH may be accurately identified and categorized by antenatal sonography. The outcome is usually poor, especially for those fetuses affected by higher-grade intraventricular hemorrhages. PMID- 12905504 TI - Examination of fetal nasal bone and repeatability of measurement in early pregnancy. AB - OBJECTIVE: To evaluate the examination and measurement of fetal nasal bone at 10 14 weeks of gestation. METHODS: The study included 501 fetuses in 496 consecutive pregnant women attending for the routine first-trimester ultrasound examination. The presence or absence of the fetal nasal bone was determined in the mid sagittal plane and the length was measured by one of four examiners (measurement A; n = 501). A second measurement was taken by the same examiner (B, n = 300) and a different examiner repeated the measurement (C, n = 200) whenever possible. Measurements were made to the nearest 0.1 mm. The duration of one hundred consecutive examinations was recorded, as was that of another 100 consecutive routine first-trimester examinations without measuring the nasal bone. RESULTS: The median nasal bone length was 1.6 (0.8-2.4) mm, the median gestational age was 12 (10-14) weeks and the median crown-rump length (CRL) was 63 (32-90) mm. The fetal profile was examined in all 501 cases and the fetal nasal bone was present in all but one case (99.8%). No transvaginal scans were needed for the examination of nasal bone only. The average time for the sonographic examination (8.3 min) was not significantly different from the average time for first trimester scans in which the fetal nasal bone was not measured (8.0 min). The fetal nasal bone length increased from 1.1 mm at a CRL of 35 mm to 2.1 mm at a CRL of 90 mm (nasal bone = 0.016 x CRL + 0.619, P < 0.001, r = 0.655). The repeatability coefficient for intraobserver measurements was 0.080 mm and the coefficient for interobserver measurements was similar (0.083 mm). CONCLUSIONS: The nasal bone can be detected from 10 weeks of gestation onwards. Consistent visualization and repeatable measurement of fetal nasal bone can be performed by an experienced sonographer in the first trimester without extending the length of time required for scanning or introducing the need for transvaginal sonography. PMID- 12905505 TI - Learning curve for sonographic examination of the fetal nasal bone at 11-14 weeks. AB - OBJECTIVE: To determine the number of ultrasound examinations necessary for training sonographers to examine accurately the fetal nasal bone at 11-14 weeks' gestation. METHODS: Fifteen sonographers with experience in measuring nuchal translucency were asked to examine the nasal bone during the routine 11-14-week scan. The supervising doctor recorded if the sonographer succeeded in obtaining the correct image. Each sonographer performed a total of 140 examinations, and the data were analyzed in seven groups of 20 examinations. In a second study, two sonographers with extensive experience in examining the nasal bone examined independently 100 consecutive patients at a median fetal crown-rump length of 65 (45-84) mm and median gestational age of 12 (11-14) weeks and recorded whether the nasal bone was absent or present. RESULTS: In the first group of 20 examinations, there was failure to obtain the correct image of the fetal profile in 1-5 (median, 4) cases. In the subsequent three groups, there was failure to obtain the correct image in 0-3 (median, 1) cases. In the fifth and sixth groups failure occurred in 0-2 (median, 0) cases and in the seventh group all sonographers obtained successful images of the fetal profile in all cases. One sonographer obtained successful images of all cases after the first 40 scans, four after the first 60 scans, six after the first 80 and two each after the first 100 and 120 scans. In the second study, there was agreement between the two sonographers that the nasal bone was absent in two and present in 98 of the 100 consecutive patients examined. CONCLUSION: The minimum number of scans required for an experienced sonographer to become competent in examining the fetal nasal bone is on average 80, with a range of 40-120. PMID- 12905506 TI - Intra- and interoperator variability in fetal nasal bone assessment at 11-14 weeks of gestation. AB - OBJECTIVES: Examination of the fetal nasal bones by ultrasound between 11 and 14 weeks of gestation has been proposed as an additional tool in the detection of trisomy 21 in a high-risk population. However the variability in the identification of fetal nasal bones by ultrasound has not yet been investigated. The aim of this study was to assess the intraobserver and interobserver reproducibility of fetal nasal bone identification by ultrasound between 11 and 14 weeks of gestation. METHODS: A total of 1040 consecutive ultrasound examinations were performed at 11-14 weeks of gestation for nuchal translucency (NT) measurement and nasal bone identification by ultrasound. A total of 657 consecutive video-loops were assessed by three experienced operators. Each operator assigned cases to one of three categories, namely present, uncertain or absent nasal bones, and the results were compared between operators. To assess the intraoperator variability each operator reviewed 100 randomly selected videos out of the 657 loops and again used the same classification. Results were compared by pairwise unweighted and weighted Kappa index to evaluate the inter- and intraoperator variability. RESULTS: Among the 1040 fetuses, there were 51 (4.9%) with an NT measurement above the 95th centile. Nasal bones were identified by ultrasound in 948, not seen in eight and impossible to assess in 84 fetuses. Four fetuses had trisomy 21 including three with absent nasal bones and increased NT and one with present nasal bones and normal NT. The Kappa and weighted Kappa values for interoperator variability between the three operators were between 0.26 and 0.37 and 0.33 and 0.44, respectively. The Kappa and weighted Kappa values for intraoperator variability were between 0.35 and 0.48 and 0.43 and 0.55, respectively. CONCLUSION: The assessment of fetal nasal bones is only fairly reproducible. Although the performance of the test in fetuses at high risk for trisomy 21 has been reported to be good, its implementation as an additional screening technique in the general population must be accompanied by teaching and quality control programs. PMID- 12905507 TI - Delta-NT or NT MoM: which is the most appropriate method for calculating accurate patient-specific risks for trisomy 21 in the first trimester? AB - OBJECTIVE: To assess whether in screening for trisomy 21 by nuchal translucency (NT) the delta or the multiples of the median (MoM) approach is the most appropriate method for calculating accurate individual patient-specific risks. METHODS: Data on fetal NT and crown-rump length from 128,030 unaffected and 428 trisomy 21 pregnancies, measured by sonographers who had obtained The Fetal Medicine Foundation Certificate of Competence in the 11-14-Week Scan, were used. We examined first, if the distribution of NT MoM and log(10)(NT MoM) was Gaussian; second, if the standard deviation of the distributions did not change with gestation; and third, if the median MoM in the affected population was a constant proportion of the median for unaffected pregnancies. All of these features are required to underpin the MoM approach. NT distributions and those of delta-NT were also analyzed. A non-parametric kernel density method was then used to assess the validity of both methods. Errors in the estimation of individual patient-specific risks using the MoM approach were assessed. RESULTS: In the unaffected pregnancies, the distributions of NT MoM and log(10)(NT MoM) were not Gaussian and the standard deviation of log(10)(NT MoM) decreased with gestation. In the trisomy 21 pregnancies, the median NT MoM decreased significantly with gestation, whereas the median delta-NT did not change with gestation. The non parametric density approach showed that contours of constant likelihood ratio were parallel to the gestational age-dependent median NT values, thus supporting the delta-NT approach. The NT MoM approach resulted in women being given an overestimate of risk for trisomy at 11 weeks and a considerable underestimate of risk at 13 weeks. CONCLUSION: In the calculation of risk for trisomy 21 by NT the NT MoM approach is inaccurate and inappropriate because the underlying assumptions are not valid. In contrast, the delta-NT approach gives accurate estimates of risks. PMID- 12905508 TI - First-trimester screening for aneuploidy with fetal nuchal translucency in a United States population. AB - OBJECTIVE: To examine the detection rate of chromosomal abnormalities using a combination of nuchal translucency (NT) and maternal age in a United States population. METHODS: A total of 2131 pregnancies with 2339 fetuses underwent NT screening from April 2000 to April 2002 in our ultrasound unit. Nuchal translucency was measured from 11 to 14 weeks' gestation. Fetal crown-rump length (CRL) was also measured. The risk for trisomy 21 was calculated from a combination of maternal age and fetal NT with the use of software provided by The Fetal Medicine Foundation (FMF). Sensitivity and false-positive rates were calculated for different risk cut-offs. RESULTS: Chromosomal defects were diagnosed in 32 cases, including 12 cases of trisomy 21 and 10 cases of trisomy 18. The estimated risk based on maternal age and fetal NT was 1 in 300 or greater in 195 (8.3%) cases and these included 10/12 (83.3%) pregnancies with trisomy 21 and 9/10 (90.0%) pregnancies with trisomy 18. CONCLUSION: A combination of maternal age and fetal NT provides an effective method of screening for chromosomal defects. Using ultrasound techniques and risk algorithms from The FMF, the performance of the test in an American population is similar to that described in international populations. PMID- 12905509 TI - Validity of the Hewlett-Packard actograph in detecting fetal movements. AB - OBJECTIVE: To detect fetal movements during non-stressed cardiotocography to evaluate the validity of the Hewlett-Packard Doppler ultrasound actograph in detecting fetal movements. METHODS: This was a prospective, observational study. Thirty healthy pregnant women were divided into two gestational age groups (Group I, 31 + 0 to 34 + 0 weeks, n = 15; Group II, 37 + 0 to 40 + 0 weeks, n = 15). A Hewlett-Packard M-1350-A actocardiograph was used to make recordings of 45 min for each woman. Fetal movements were also detected sonographically by an observer and they were recorded independently by the pregnant woman. Ninety consecutive 30 s periods were analyzed to study agreement between the three techniques. RESULTS: The mean total agreement for fetal activity plus quiescence between ultrasound and the actograph was 63.7% for Group I, and 72.2% for Group II. In cases of fetal activity only, these values were 45.8% and 47.6%, respectively. Total agreements between ultrasound and the pregnant woman were 74.0% and 79.0%, respectively, for Groups I and II. Cohen's kappa values for total agreement between ultrasound and actograph were 0.31 (95% CI, 0.27-0.36), indicating 'fair' agreement in Group I, and 0.43 (95% CI, 0.38-0.47), indicating 'moderate' agreement in Group II. CONCLUSIONS: Use of the Hewlett-Packard actograph may offer some additional information during routine cardiotocography. However, our results were disappointing; the technique appears not to be suitable for detailed recordings of fetal behavior, and may even be misleading when difficult cardiotocographic patterns have to be interpreted. PMID- 12905510 TI - Is there a relationship between estimated fetal weight and umbilical artery Doppler impedance indices? AB - OBJECTIVE: To establish whether there is a relationship between estimated fetal weight (EFW) and umbilical artery Doppler waveform impedance indices in the third trimester. METHODS: The pulsatility index (PI) and S/D (systolic/diastolic) ratio were obtained together with the EFW from 274 low-risk pregnancies. Measurements were made at fortnightly intervals from 30 weeks' gestation until delivery. A relationship between the two impedance indices and EFW was sought at gestational age ranges of 30-32, 33-35, 36-38 and 39-41 weeks. RESULTS: There were 918 pairs of PI (with S/D) and EFW available for analysis. The mean of the impedance indices decreased with advancing gestational age as expected. There was no clinically relevant correlation between impedance indices and EFW within any of the gestational age ranges. CONCLUSION: Impedance indices from the umbilical artery Doppler waveform decrease with advancing gestational age due, at least partially, to expansion of the placental vascular tree. Within narrow gestational age ranges in the third trimester, there is no meaningful correlation between fetal weight and impedance indices. It is therefore not necessary to adjust umbilical artery Doppler impedance indices to account for fetal size. PMID- 12905511 TI - Cross-sectional and longitudinal evaluation of uterine artery Doppler velocimetry for the prediction of pre-eclampsia in normotensive women with specific risk factors. AB - OBJECTIVE: To evaluate the performance, in the prediction of pre-eclampsia, of (1) an abnormal mean uterine artery resistance index (RI; cross-sectional index) at 24 weeks of gestation, (2) the individual longitudinal flow pattern of results observed at 16, 20 and 24 weeks of gestation and (3) a multiple logistic regression model including the individual longitudinal flow pattern and the mean RI at 24 weeks. METHODS: A total of 144 normotensive pregnant women with risk factors for pre-eclampsia were evaluated with uterine artery color Doppler at 16, 20 and 24 weeks' gestation. The following indices were obtained: (1) cross sectional index: the mean RI of both uterine arteries at 24 weeks' gestation was calculated for each patient and considered abnormal when >/= 0.58; (2) longitudinal indices: the individual longitudinal flow pattern of mean RI of both the main uterine arteries at 16, 20 and 24 weeks' gestation was derived for each subject and defined as (a) the typical physiological flow pattern, with a trend of decrease in values or (b) the non-physiological flow pattern. The probability of having a pregnancy complicated by pre-eclampsia was also calculated by means of a multivariate logit model. The log-odds was modeled as a function of variables related to maternal characteristics, the individual longitudinal flow pattern indicator, and of the mean RI at 24 weeks' gestation as a continuous variable. RESULTS: Pregnancies had a physiological course in 108 (75%) women, while 36 (25%) women developed pre-eclampsia during the third trimester. For the prediction of pre-eclampsia, the use of an abnormal uterine artery RI index (> or = 0.58) at 24 weeks showed a sensitivity of 77.8%, a specificity of 67.6%, a positive predictive value (PPV) of 44.4% and a negative predictive value (NPV) of 90.1%, with a likelihood ratio (LR) for an abnormal test of 2.4; with the longitudinal flow pattern indicator, sensitivity was 88.9%, specificity 82.4%, PPV 62.7% and NPV 95.7%, with a LR for an abnormal test of 4.9; the use of a logit model yielded a sensitivity of 72.2%, a specificity of 90.7%, a PPV of 72.2% and a NPV of 90.7%, with a LR for an abnormal test of 8.0. CONCLUSIONS: In this study the use of an individual longitudinal flow pattern indicator resulted in improving accuracy in the prediction of pre-eclampsia as compared with the traditional cross-sectional mean RI at 24 weeks. A further increase in specificity and PPV was obtained using a logit model that includes the longitudinal flow pattern indicator and the cross-sectional RI at 24 weeks. Since both the longitudinal flow pattern indicator and the logit model showed a high performance in predicting pre-eclampsia in women with risk factors for impaired placentation, they might be used to identify a high-risk population in which preventive measures and/or therapeutic options might be tested. PMID- 12905512 TI - The pedicle artery sign based on sonography with color Doppler imaging can replace second-stage tests in women with abnormal vaginal bleeding. AB - OBJECTIVE: Unenhanced transvaginal sonography is not accurate in the detection of endometrial polyps. Currently, second-stage tests such as saline contrast sonohysterography and office hysteroscopy are used to diagnose endometrial lesions, but both have limitations and side effects. We proposed visualization of the pedicle artery on color Doppler imaging as a sign of polyps. METHODS: This was a prospective observational study involving 3099 consecutive patients referred for assessment of the endometrium and myometrium. The uterus was assessed in sagittal and coronal planes using color Doppler sonography. Patients with suspected endometrial pathology on sonography were noted and the volume of the focal lesion was calculated. Patients with a clearly visible pedicle artery reaching the central part of the endometrium were regarded as test-positive. The gold standard was defined as the presence or absence of an endometrial polyp at hysterectomy, hysteroscopy and/or endometrial histology on dilatation and curettage within 1 year of sonography. RESULTS: Of the 3099 women, no gold standard was available in 2230. Only 28 of the 199 patients who were test positive did not have a gold standard. In the 869 patients in whom a gold standard was available, 182 had one or more endometrial polyps. The pedicle artery test had an apparent sensitivity for detection of endometrial polyps of 76.4%, specificity of 95.3%, positive predictive value (PPV) of 81.3%, and negative predictive value of 93.8%. When extending the test to the prediction of any focal intracavitary pathology the PPV was 94.2%. CONCLUSIONS: The pedicle artery test has a very high PPV for intracavitary pathology. We expect that in the majority of patients with an endometrial polyp this test may replace more invasive established second-stage tests, such as saline contrast sonohysterography and office hysteroscopy. PMID- 12905513 TI - Pelvic ultrasound evaluation in patients with Turner syndrome during treatment with growth hormone. AB - OBJECTIVES: Treatment with growth hormone (GH), alone or in combination with oxandrolone, is used in patients affected by Turner syndrome to improve growth velocity and adult height. Since GH interacts with gonadotropins in the stimulation of the human ovary, the aim of our study was to evaluate the possible effects of GH administration on uterine and ovarian characteristics. METHODS: We performed pelvic ultrasound assessment in 29 patients with Turner syndrome aged 7.5-16.6 years (19 with 45,X karyotype; 10 with variant karyotypes) before and during treatment with GH alone. Uterine volume and ovarian size and morphology were compared to those of 23 age-matched girls with Turner syndrome not treated with GH. Both patients and controls were divided into prepubertal and pubertal groups. Cross-sectional and longitudinal studies (before and every 6 months during GH treatment for 2 years) were performed. RESULTS: We observed a significantly higher uterine anteroposterior diameter and volume in younger (< or = 11 years) GH-treated Turner syndrome girls than in those who were untreated. Also visualization and heterogeneous echopattern of the ovaries were significantly more frequent in treated than in untreated Turner syndrome patients, particularly before the age of 11 years. The longitudinal study showed a significant increase in uterine volume, more related to treatment than to age. Spontaneous breast development and menarche were found more frequently in GH treated Turner syndrome girls. CONCLUSION: Growth hormone therapy can have a co gonadotropin role in patients with Turner syndrome. PMID- 12905514 TI - Fetal cerebellar hemorrhage in a severely growth-restricted fetus: natural history and differential diagnosis from Dandy-Walker malformation. AB - This is a report of an intracerebellar hemorrhage in a severely growth-restricted fetus with pathological Doppler findings of the fetal and uteroplacental circulations. The diagnosis was made sonographically at 22 weeks of gestation and the natural course of the hemorrhage was followed. Interestingly, the final sonographic appearance of the posterior fossa was quite similar to that of the classic form of Dandy-Walker malformation: absence of the vermis and an enlarged fourth ventricle. However, careful sonographic examination showed that the enlargement of the fourth ventricle was actually caused by a porencephalic cystic lesion of the left cerebellar lobe. Pathological examination revealed complete absence of the vermis and cerebellar hypoplasia. PMID- 12905515 TI - Transient hydrops fetalis of the donor fetus in twin-twin transfusion syndrome after therapeutic amnioreduction. AB - We present a case of twin-twin transfusion syndrome associated with transient hydrops fetalis observed in the donor after therapeutic amnioreduction at 22 weeks of gestation. After the amnioreduction, the bladder of the donor could be visualized and the donor subsequently began to make amniotic fluid, with spontaneous regression of hydrops fetalis. It is suspected that after therapeutic amnioreduction, intrauterine hemodynamic changes occurred and the donor developed transient hydrops fetalis due to volume overload. PMID- 12905516 TI - Doppler sonography for predicting fetal anemia caused by massive fetomaternal hemorrhage. AB - Fetomaternal hemorrhage (FMH) can cause severe anemia in the fetus. Untreated, this may cause hydrops or even fetal death. However, correct diagnosis of FMH followed by blood transfusion can prevent these life-threatening consequences. We describe two cases in which fetal anemia was suspected because of maternal reporting of decreased or absent fetal movements, the detection of a sinusoidal heart rate pattern and increased blood flow velocities of the middle cerebral artery and umbilical vein. Together with the Betke-Kleihauer test showing fetal cells in the maternal circulation, this led to the correct diagnosis of severe fetal anemia caused by FMH. A Cesarean section was performed within a few hours. Both neonates were severely anemic and received immediate blood transfusions. They are currently alive and well. PMID- 12905517 TI - Prenatal sonographic diagnosis of isolated agnathia: a case report. AB - Agnathia is a rare malformation characterized by the absence of the mandible, microstomia, aplasia or hypoplasia of the tongue, and low-set or medially fused ears. It occurs alone or in combination with a variety of malformations such as holoprosencephaly. All previous cases of the prenatal diagnosis of agnathia have been accompanied with fatal anomalies. We report here the first case of isolated agnathia, which was diagnosed by prenatal sonography at 32 weeks of gestation. In this case, prenatal sonographic examination showed the severe defect of the chin, small opening of the mouth and severe polyhydramnios. There was no sonographic evidence of holoprosencephaly or anomalies in other organ systems. The diagnosis was confirmed postnatally. PMID- 12905518 TI - Myometrial trophoblastic implant as a complication of surgically induced first trimester termination of pregnancy. AB - Persistent trophoblastic tissue has been described in the abdominal cavity after surgical treatment of tubal ectopic pregnancy. More infrequently the cause of the ectopic trophoblast is linked to uterine perforation due to surgically induced termination of pregnancy (TOP). Ultrasonographic images may suggest an ectopic pregnancy. A case of myometrial trophoblastic tissue implantation following surgically induced first-trimester TOP is described. PMID- 12905520 TI - Conjoined twins in a triplet pregnancy: early prenatal diagnosis with three dimensional ultrasound and review of the literature. AB - We report two cases of conjoined twins complicating a triplet pregnancy diagnosed by two-dimensional ultrasound in the first trimester and evaluated further by three-dimensional ultrasound. A review of the literature over the last 30 years revealed 11 other cases diagnosed prenatally by ultrasound. Overall, three (23%) of these 13 pregnancies were achieved by assisted reproductive techniques and 10 (77%) were diagnosed before 18 weeks. Four women opted for termination of the whole pregnancy and three were managed expectantly, with two delivering before 32 weeks. Two monochorionic pregnancies underwent selective feticide with intrauterine demise of the non-conjoined fetus in both cases. All four dichorionic pregnancies undergoing selective termination or spontaneous embryo reduction to singleton in the first trimester resulted in term delivery of the non-conjoined fetus. The rare condition of conjoined twins in a triplet pregnancy poses a significant obstetric challenge from both diagnostic and management points of view. Accurate determination of chorionicity in these cases plays a critical role in determining management and outcome. PMID- 12905519 TI - Pelvic echinococcus mimicking multicystic ovary. AB - An unusual case of pelvic echinococcus cyst is presented, appearing initially on transvaginal ultrasound as a pelvic mass mimicking a multicystic ovary. A similar mass in the liver raised preoperatively the suspicion of echinococcosis, making an open surgical procedure preferable to laparoscopy. Diagnosis was confirmed pathologically after removal of the cyst. Though their location in the pelvis is rare, echinococcal cysts should be considered in the differential diagnosis of pelvic masses, especially in patients from endemic areas. Evaluation of previous medical history and current symptoms, together with the ultrasonographic findings, is important for correct diagnosis and appropriate management. PMID- 12905521 TI - Introital and transvaginal ultrasound as the main tool in the assessment of urogenital and pelvic floor dysfunction: an imaging panel and practical approach. AB - This article reviews the different applications of ultrasound in benign urogynecological diseases. The findings presented here were obtained by introital and transvaginal ultrasound, both of which can be performed with the same equipment (5-7-MHz sector transducer, emission angle of at least 90 degrees; for introital sonography, the transducer is placed over the external urethral orifice with the transducer axis corresponding to the body axis). Female voiding dysfunction, including urge symptoms, recurrent urinary tract infections and urinary incontinence, may occur secondary to morphological and topographical changes of the urogenital organs. Findings such as urethral diverticula, periurethral masses, funneling of the urethra and distension cystoceles are identified by introital ultrasound. Transvaginal ultrasound enables the detection of pathologies of the bladder and uterus including its appendages. Ultrasound as part of the diagnostic work-up of stress urinary incontinence and genitourinary prolapse allows for the morphological and dynamic assessment of the lower urinary tract. It is possible, for example, to classify sonographically identified changes of the endopelvic fascia as lateral (distraction cystocele, funneling of the urethra) and central (pulsation cystocele) defects as well as to determine the reactivity of the pelvic floor muscles. Ultrasound has replaced radiography in yielding information on the abnormal morphology of the urogenital organs, which should be taken into account in planning the treatment of urogynecological conditions. PMID- 12905522 TI - Prenatal diagnosis of otocephaly with microphthalmia/anophthalmia using ultrasound and magnetic resonance imaging. PMID- 12905523 TI - Detecting anal sphincter injury: acceptability and feasibility of endoanal ultrasound immediately postpartum. PMID- 12905524 TI - The 'war' of logistic regression models in the diagnosis of ovarian cancer. PMID- 12905527 TI - Wild type p53 gene causes reorganization of cytoskeleton and, therefore, the impaired deformability and difficult migration of murine erythroleukemia cells. AB - We studied the role of p53 gene in the biophysics and biology in murine erythroleukemia cell line (MEL), with the goal of understanding the influence of this tumor suppressor gene on the deformability and metastasis of tumor cells. Experiments were performed on MEL and p53-transfected MEL (MEL-M with mutant p53 gene and MEL-W with wild-type p53 gene). The cell growth curves indicated that the over-expression of wild-type p53 gene significantly suppressed the growth of MEL, with G(0)-G(1) arrest and apoptosis shown by flow cytometric assays. Confocal laser scanning microscopy revealed that the MEL-W had a more compact organization of the F-Actin cytoskeleton than MEL and MEL-M. Fluorescence polarization measurement indicated a higher membrane fluidity of MEL-W than the other two groups. Fourier transform infrared spectroscopy (FT-IR) showed changes in the composition and/or structure of membrane lipids in MEL-W, with decreases in secondary structures of proteins such as alpha-helix, turns and bends and random coil, in comparison to MEL and MEL-M. The osmotic fragility curves indicated that MEL-W was more fragile and micropipette experiments showed that they had increased elasticity and reduced deformability in comparison to MEL and MEL-M. The adhesion assay with the use of the flow chamber revealed a lower adhesion rate of MEL-W to endothelial cells at high shear stress. The present study on the molecular biology with biophysics of MEL cells contributes to our knowledge on the tumor suppressor gene p53. PMID- 12905528 TI - Cytokeratin intermediate filament organisation and dynamics in the vegetal cortex of living Xenopus laevis oocytes and eggs. AB - Cytokeratin intermediate filaments are prominent constituents of developing Xenopus oocytes and eggs, forming radial and cortical networks. In order to investigate the dynamics of the cortical cytokeratin network, we expressed EGFP tagged Xenopus cytokeratin 1(8) in oocytes and eggs. The EGFP-cytokeratin co assembled with endogenous partner cytokeratin proteins to form fluorescent filaments. Using time-lapse confocal microscopy, cytokeratin filament assembly was monitored in live Xenopus oocytes at different stages of oogenesis, and in the artificially-activated mature egg during the first cell cycle. In stage III to V oocytes, cytokeratin proteins formed a loose cortical geodesic network, which became more tightly bundled in stage VI oocytes. Maturation of oocytes into metaphase II-arrested eggs induced disassembly of the EGFP-cytokeratin network. Imaging live eggs after artificial activation allowed us to observe the reassembly of cytokeratin filaments in the vegetal cortex. The earliest observable structures were loose foci, which then extended into curly filament bundles. The position and orientation of these bundles altered with time, suggesting that forces were acting upon them. During cortical rotation, the cytokeratin network realigned into a parallel array that translocated in a directed manner at 5 microm/minute, relative to stationary cortex. The cytokeratin filaments are, therefore, moving in association with the bulk cytoplasm of the egg, suggesting that they may provide a structural role at the moving interface between cortex and cytoplasm. PMID- 12905529 TI - Human polymorphonuclear leukocytes respond to waves of chemoattractant, like Dictyostelium. AB - It has been assumed that the natural chemotactic signal that attracts human polymorphonuclear leukocytes (PMNs) over long distances to sites of infection is in the form of a standing spatial gradient of chemoattractant. We have questioned this assumption on the grounds, first, that standing spatial gradients may not be stable over long distances for long periods of time and, second, that in the one animal cell chemotaxis system in which the natural chemotactic signal has been described in space and time, aggregation of Dicytostelium discoideum, the signal is in the form of an outwardly relayed, nondissipating wave of attractant. Here, it is demonstrated that PMNs alter their behavior in each of the four phases of a wave of PMN chemoattractant, fashioned after the Dictyostelium wave, in a manner similar to Dictyostelium. These results demonstrate that PMNs have all of the machinery to respond to a natural wave of attractant, providing support to the hypothesis that the natural signal that attracts PMNs over large distances to sites of infection in the human body may also be in the form of a wave. PMID- 12905530 TI - Expression of class III beta-tubulin reduces microtubule assembly and confers resistance to paclitaxel. AB - Human brain and testis specific betaIII-tubulin was amplified from a cDNA library, modified to encode a C-terminal hemagglutinin antigen epitope tag, and cloned into a vector that allows tetracycline regulated expression in mammalian cells. Immunofluorescence analysis of transfected Chinese hamster ovary cells demonstrated that expressed HA-tagged betaIII-tubulin is able to assemble with endogenous tubulin into microtubules even though betaIII-tubulin is not a normal constituent of these cells. A stable G418-resistant clone with moderate HAbetaIII tubulin expression displayed weak (1.5-2-fold) resistance to paclitaxel. A second clone with higher HAbetaIII-tubulin expression could not grow unless tetracycline was present to repress transcription of the transfected cDNA. Analysis of cellular microtubules in each of these clones indicated that incorporation of HAbetaIII-tubulin led to a significant expression-dependent decrease in assembled tubulin. Paclitaxel resistant cells were also directly selected from the transfected cell population using a paclitaxel concentration 4 times higher than the minimum toxic dose. Few cells were able to survive the selection and they grew very slowly. Western blot analysis of these resistant cells revealed very high HAbetaIII-tubulin expression that led to almost complete replacement of endogenous beta-tubulin at steady state. Transfected betaIII-tubulin with no epitope tag behaved in a very similar fashion indicating that presence of the HA tag had no discernible functional effect. The results demonstrate that betaIII tubulin diminishes microtubule assembly, is toxic when present at high levels, but is able to confer weak resistance to paclitaxel when expressed at moderate levels in mammalian cells. PMID- 12905532 TI - Fossil sister group of craniates: predicted and found. AB - This study investigates whether the recently described Cambrian fossil Haikouella (and the very similar Yunnanozoon) throws light on the longstanding problem of the origin of craniates. In the first rigorous cladistic analysis of the relations of this animal, we took 40 anatomical characters from Haikouella and other taxa (hemichordates, tunicates, cephalochordates, conodont craniates and other craniates, plus protostomes as the outgroup) and subjected these characters to parsimony analysis. The characters included several previously unrecognized traits of Haikouella, such as upper lips resembling those of larval lampreys, the thick nature of the branchial bars, a mandibular branchial artery but no mandibular branchial bar, muscle fibers defining the myomeres, a dark fibrous sheath that defines the notochord, conclusive evidence for paired eyes, and a large hindbrain and diencephalon in the same positions as in the craniate brain. The cladistic analysis produced this tree: (protostomes, hemichordates (tunicates, (cephalochordates, (Haikouella, (conodonts + other craniates))))), with the "Haikouella + craniate" clade supported by bootstrap values that ranged from 81-96%, depending on how the analysis was structured. Thus, Haikouella is concluded to be the sister group of the craniates. Alternate hypotheses that unite Haikouella with hemichordates or cephalochordates, or consider it a basal deuterostome, received little or no support. Although it is the sister group of craniates, Haikouella is skull-less and lacks an ear, but it does have neural crest derivatives in its branchial bars. Its craniate characters occur mostly in the head and pharynx; its widely spaced, robust branchial bars indicate it ventilated with branchiomeric muscles, not cilia. Despite its craniate mode of ventilation, Haikouella was not a predator but a suspension feeder, as shown by its cephalochordate-like endostyle, and tentacles forming a screen across the mouth. Haikouella was compared to pre-craniates predicted by recent models of craniate evolution and was found to fit these predictions closely. Specifically, it fits Northcutt and Gans' prediction that the change from ciliary to muscular ventilation preceded the change from suspension feeding to predatory feeding; it fits Butler's claim that vision was the first craniate sense to start elaborating; it is consistent with the ideas of Donoghue and others about the ancestor of conodont craniates; and, most strikingly, it resembles Mallatt's prediction of the external appearance of the ancestral craniate head. By contrast, Haikouella does not fit the widespread belief that ancestral craniates resembled hagfishes, because it has no special hagfish characters. Overall, Haikouella agrees so closely with recent predictions about pre-craniates that we conclude that the difficult problem of craniate origins is nearly solved. PMID- 12905531 TI - Changes in the abundance and distribution of conserved centrosomal, cytoskeletal and ciliary proteins during spermiogenesis in Marsilea vestita. AB - Spermiogenesis in the male gametophytes of the water fern Marsilea vestita is a precise and rapid process resulting in the production of ciliated gametes. Development begins from a single cell within the microspore wall that undergoes nine rapid cell division cycles in distinct planes to produce 32 spermatids that are surrounded by 7 sterile cells. Thereafter, the de novo formation of basal bodies occurs in a discrete cytoplasmic particle known as a blepharoplast, with the subsequent formation of a complex ciliary apparatus in elongating spermatids. The rate and extent of development appear to be controlled at a post transcriptional level, where the sudden translation of specific stored mRNAs (e.g., centrin) results in the formation of particular structures in the cells (e.g., blepharoplasts). We show here that additional centrosomal and cytoskeletal antigens known as SF assemblin, p95 kDa protein, delta tubulin, gamma tubulin, Xgrip109, Aik, CTR453, RanBPM, BX63, RSP6, and alpha tubulin each exhibit specific localization patterns both on immunoblots of gametophyte protein isolates and in fixed cells. BAp90, PP4, and RLC exhibit specific localization patterns in fixed cells. We show that the antigens exhibit complex patterns of abundance during spermiogenesis. In an attempt to identify regulatory agents involved in spermiogenesis, we employed a RNAi-based screen of 41 randomly selected gametophyte cDNAs on developing populations of synchronously growing gametophytes. The gametophytes treated with each of the RNAi probes exhibited arrest at a specific stage of development. A consequence of anomalous development was the block to assembly of the ciliary apparatus, an effect highlighted by altered staining with anti-centrin, anti-beta-tubulin, and anti-RSP6 antibodies. Our results show that complex, integrated processes of translation and protein partitioning apparently underlie the assembly of the ciliary apparatus during spermiogenesis in male gametophytes of M. vestita. PMID- 12905533 TI - Cranial osteology in Momotidae (Aves: Coraciiformes). AB - Momotidae (motmots) is found throughout Latin America between Mexico and northern Argentina. Given the absence of detailed studies of cranial osteology of Momotidae in the literature, this article presents a comprehensive description of the variation of the cranial osteology in all nine species of Momotidae and compares the results with published studies of other families of Coraciiformes and families in other orders. In addition, the cranial structures described are related to ecological and behavioral aspects of Momotidae. The cranial osteology of Baryphthengus ruficapillus is described in detail and compared with other species of Momotidae. The results indicate the presence in Momotidae of modified cranial structures, among which the most conspicuous are the frontal, lacrimal, squamosal, orbital, and laterosphenoid regions, as well as the palatine, upper jaw, pterygoid, and mandible. PMID- 12905534 TI - Distribution of keratin and associated proteins in the epidermis of monotreme, marsupial, and placental mammals. AB - The expression of acidic and basic keratins, and of some keratinization marker proteins such as filaggrin, loricrin, involucrin, and trichohyalin, is known for the epidermis of only a few eutherian species. Using light and high-resolution immunocytochemistry, the presence of these proteins has been studied in two monotreme and five marsupial species and compared to that in eutherians. In both monotreme and marsupial epidermis lamellar bodies occur in the upper spinosus and granular layers. Development of the granular layer varies between species and regionally within species. There is great interspecific variation in the size (0.1-3.0 microm) of keratohyalin granules (KHGs) associated with production of orthokeratotic corneous tissues. Those skin regions lacking hairs (platypus web), or showing reduced pelage density (wombat) have, respectively, minute or indiscernible KHGs, associated with patchy, or total, parakeratosis. Ultrastructural analysis shows that monotreme and marsupial KHGs comprise irregular coarse filaments of 25-40 nm that contact keratin filaments. Except for parakeratotic tissues of platypus web, distribution of acidic and basic proteins in monotreme and marsupial epidermis as revealed by anti-keratin antibodies AE1, AE2, and AE3 resembles that of eutherian epidermis. Antibodies against human or rat filaggrins have little or no cross-reactivity with epidermal proteins of other mammals: only sparse areas of wombat and rabbit epidermis show a weak immunofluorescence in transitional cells and in the deepest corneous tissues. Of the available, eutherian-derived antibodies, that against involucrin shows no cross-reactivity with any monotreme and marsupial epidermal tissues and that against trichohyalin cross-reacts only with cells in the inner root sheath and medulla of hairs. These results suggest that if involucrin and trichohyalin are present throughout noneutherian epidermis, they may have species-specific molecular structures. By contrast, eutherian-derived anti-loricrin antibodies show a weak to intense cross-reactivity to KHGs and corneous tissues of both orthokeratotic and parakeratotic epidermis in monotremes and marsupials. High resolution immunogold analysis of loricrin distribution in immature keratinocytes of platypus parakeratotic web epidermis identifies labeled areas of round or irregular, electron-pale granules within the denser keratohyalin component and keratin network. In the deepest mature tissues, loricrin-like labeling is diffuse throughout the cytoplasm, so that cells lack the preferential distribution of loricrin along the corneous envelope that characterizes mature eutherian keratinocytes. Thus, the irregular distribution of loricrin in platypus parakeratotic tissues more resembles that which has been described for reptilian and avian keratinocytes. These observations on the noneutherian epidermis show that a stratum granulosum is present to different degrees, even discontinuous within one tissue, so that parakeratotic and orthokeratotic areas may alternate: this might imply that parakeratotic monotreme epidermis reflects the primitive pattern of amniote alpha-keratogenesis. Absent from anamniote epidermis and all sauropsid beta-keratogenic tissues, the ubiquitous presence of a loricrin-like protein as a major component of other amniote corneous tissues suggests that this is a primitive feature of amniote alpha-keratogenesis. The apparent lack of specific regionalization of loricin near the plasma membranes of monotreme keratinocytes could be an artifactual result of the immunofluorescence technique employed, or there may be masking of the antigenicity of loricrin-like proteins once they are incorporated into the corneous envelope. Nevertheless, the mechanism of redistribution of such proteins during maturation of monotreme keratinocytes is different from, perhaps more primitive, or less specialized, than that in the epidermis of eutherian mammals. PMID- 12905535 TI - Differential mesowear in occluding upper and lower molars: opening mesowear analysis for lower molars and premolars in hypsodont horses. AB - A new approach of reconstructing ungulate diet, the mesowear method, was recently introduced by Fortelius and Solounias ([2000] Am Mus Novitat 3301:1-36). Mesowear is based on facet development on the occlusal surfaces of the teeth. Restricting mesowear investigation to maxillary cheek teeth would allow mesowear investigation only in assemblages of large numbers of individuals and therefore would generally restrict this method to relatively few assemblages of recent and fossil ungulates. Most of the fossil, subfossil, and recent ungulate osteological assemblages that may be assigned to a single taxon have smaller numbers of individuals. This results in a demand to extend the mesowear method to further tooth positions in order to obtain stable dietary classifications of fossil taxa. The focus of this article is to test if a consistent mesowear classification is obtainable for mandibular as well as for maxillary teeth. For statistical testing, large assemblages of isolated cheek teeth of the Vallesian hipparionine horse Hippotherium primigenium and of the recent zebra Equus burchelli were employed as models. The upper tooth positions P4, M1, M2, and M3 as suggested by Kaiser and Solounias (2003) as the model for the "extended" mesowear method and the lower tooth positions P4-M3 were tested for their consistency in classification of the mesowear variables. We found a considerable shift of the mesowear signature towards the grazing edge of the mesowear continuum in lower cheek teeth. In order to adjust the signal of lower teeth to the signal of the upper teeth, a calibration factor was introduced which allowed incorporation of lower cheek teeth into the same model of mesowear investigation together with upper cheek teeth. We propose that this model is particularly suited for the reconstruction of paleodiets in hypsodont hipparionine and equine equids. We further investigated the functional relation between the mesowear profiles and the distribution of dental tissues along the course of the occlusal contact. We therefore correlated mesowear profiles with enamel distribution profiles and found the mesowear profile to be strongly controlled by the attritional environment encountered by a given apex area. The differential signal observed in cusp apex morphology between upper and lower cheek teeth was found to be more closely related to attrition by the antagonistic tooth than to the distribution of dental tissues in the tooth under consideration. The results suggest a general extension of the mesowear method of paleodiet reconstruction and a basic scenario for the evolution of anisodont dentitions. PMID- 12905536 TI - Structural peculiarities of the tubercles in the skin of the turbot, Scophthalmus maximus (L., 1758) (Osteichthyes, Pleuronectiformes, Scophthalmidae). AB - The structure of the bony tubercles of the turbot, Scophthalmus maximus (L., 1758), was examined using ground sections, microradiography, SEM, and TEM. The tubercles are small, isolated, mineralized conical plates randomly distributed in the eyed side of the body. They are composed of three layers: the outer limiting layer, the external layer, and the basal plate, which make up the thin and flat elasmoid scales of Teleostei. The main difference between regular elasmoid scales and bony tubercles lies in the organization and the growth of the basal plate. Indeed, the conical shape of the tubercle is the result of a prominent thickening of the central part of the basal plate where the collagen matrix is organized in a complicated three-dimensional network. Densely packed thick collagen fibrils form superimposed plies organized in a plywood-like structure that resembles that of the elasmoid scales but it is criss-crossed by numerous vertical sheets of thin collagen fibrils. The tubercles originate from thin and flat plates located in the skin of larvae and juveniles, whose structure is that of regular developing elasmoid scales. Thus, the tubercles of Scophthalmus maximus could be considered as modified elasmoid scales rather than bony structures. They might be the result of specific arrangements related to the general trend of reduction of the dermal skeleton in the teleostean lineage. PMID- 12905537 TI - Formation of cartilage in the heart of the Spanish terrapin, Mauremys leprosa (Reptilia, Chelonia). AB - Cartilaginous deposits are regularly present in the heart of several reptilian, avian, and mammalian species. The formation of these extraskeletal cartilages has been studied in birds and mammals, but not in reptiles. The aim here was to elucidate this question in the Spanish terrapin. Hearts from 23 embryos belonging to Yntema (1968) developmental stages 17 to 26 and eight terrapins age 3 months to 10 years were examined using histological, histochemical, and immunohistochemical techniques. In the heart of the Spanish terrapin (Mauremys leprosa), chondrogenesis can start during embryonic life. Cartilaginous tissue develops from a mesenchymal cellular condensation that extends along the aorticopulmonary septum and the incipient pars fibrosa of the ventricular horizontal septum. This cellular condensation, which is smooth muscle alpha-actin (SMalpha-actin)-negative and type II collagen-negative during stages 17 to 22, acts as a prechondrogenic condensation. In stage 23, production of type II collagen begins in the central core of the condensation and gradually spreads toward its periphery. The type II collagen-positive (chondrogenic) cellular condensation remains devoid of perichondrium prior to birth. Thereafter, it converts into hyaline cartilage that extends along the proximal part of the aorticopulmonary septum and the pars fibrosa of the horizontal septum. Our findings are consistent with the assumption that, as in birds and mammals, the precursors of the cardiac chondrocytes in chelonians are neural crest-derived cells of nonmuscular nature. In addition, they point to the possibility that cells from the neural crest populate the embryonic pars fibrosa of the horizontal septum, thereby contributing to its alignment with the aorticopulmonary septum. In the present species, a second cartilaginous deposit of a hyaline nature extends along the sinus wall of the right semilunar valve of the right aorta, penetrating the fibrous cushion that constitutes the proximal support of the corresponding valve leaflet. This cartilage develops after birth, between the third and eighteenth month of life; its morphogenetic origin is unclear. The cartilaginous foci occurring in hearts of Spanish terrapin appear to act as pivots resisting mechanical tensions generated during the cardiac cycle. In the specimens examined there was no sign of replacement of the cardiac cartilages by bone tissue. PMID- 12905538 TI - Architectural properties of distal forelimb muscles in horses, Equus caballus. AB - Articular injuries in athletic horses are associated with large forces from ground impact and from muscular contraction. To accurately and noninvasively predict muscle and joint contact forces, a detailed model of musculoskeletal geometry and muscle architecture is required. Moreover, muscle architectural data can increase our understanding of the relationship between muscle structure and function in the equine distal forelimb. Muscle architectural data were collected from seven limbs obtained from five thoroughbred and thoroughbred-cross horses. Muscle belly rest length, tendon rest length, muscle volume, muscle fiber length, and pennation angle were measured for nine distal forelimb muscles. Physiological cross-sectional area (PCSA) was determined from muscle volume and muscle fiber length. The superficial and deep digital flexor muscles displayed markedly different muscle volumes (227 and 656 cm3, respectively), but their PCSAs were very similar due to a significant difference in muscle fiber length (i.e., the superficial digital flexor muscle had very short fibers, while those of the deep digital flexor muscle were relatively long). The ulnaris lateralis and flexor carpi ulnaris muscles had short fibers (17.4 and 18.3 mm, respectively). These actuators were strong (peak isometric force, Fmax=5,814 and 4,017 N, respectively) and stiff (tendon rest length to muscle fiber length, LT:LMF=5.3 and 2.1, respectively), and are probably well adapted to stabilizing the carpus during the stance phase of gait. In contrast, the flexor carpi radialis muscle displayed long fibers (89.7 mm), low peak isometric force (Fmax=555 N), and high stiffness (LT:LMF=1.6). Due to its long fibers and low Fmax, flexor carpi radialis appears to be better adapted to flexion and extension of the limb during the swing phase of gait than to stabilization of the carpus during stance. Including muscle architectural parameters in a musculoskeletal model of the equine distal forelimb may lead to more realistic estimates not only of the magnitudes of muscle forces, but also of the distribution of forces among the muscles crossing any given joint. PMID- 12905539 TI - Staging of rectal cancer. PMID- 12905540 TI - Controlling the expansion of abdominal aortic aneurysms. PMID- 12905541 TI - Origin of symptoms in diverticular disease. AB - BACKGROUND: A significant number of patients with colonic diverticula experience unexplained, recurrent, short-lived but often debilitating abdominal pain and alteration in bowel habit. Such patients account for many medical consultations every year but, as our understanding of the cause of their symptoms remains imperfect, treatment options are limited. This article reviews the possible mechanisms that may be responsible for the symptoms of diverticular disease. METHODS: Medline and Science Citation Index searches were performed to locate English language articles relating to colonic diverticula and symptoms published between January 1966 and July 2002. Manual cross-referencing was also performed and some historical articles were included. RESULTS AND CONCLUSION: Several theories now exist about the mechanisms underlying the symptoms of diverticular disease. Initial studies suggested that they may be due to alterations in the intracolonic pressure, extrapolating earlier thoughts on the likely pathogenesis of diverticula. It seems more likely, however, that several inter-related processes, such as muscular dysfunction, visceral hypersensitivity and inflammation, are involved in symptom generation. PMID- 12905542 TI - Salvage surgery after restorative proctocolectomy. AB - BACKGROUND AND METHOD: Restorative proctocolectomy is now the elective surgical procedure of choice for most patients with ulcerative colitis or familial adenomatous polyposis. Complications may lead to failure, defined as removal of the reservoir with establishment of a permanent ileostomy or long-term diversion. Failure may be avoided for some patients by salvage surgery. The causes of failure are identified in this article and the procedures adopted to treat them are defined; a review of the literature was carried out to determine the effectiveness of the procedures. RESULTS: Failure after restorative proctocolectomy results from complications, which may occur indefinitely during follow-up to a cumulative rate of about 15 per cent at 10-15 years. Sepsis accounts for over 50 per cent of these complications. Abdominal salvage procedures are successful in 20 to over 80 per cent of patients but the rate of salvage is dependent on the duration of follow-up, which might explain this variance. Local procedures are successful in 50-60 per cent of patients with pouch-vaginal fistula. Poor function accounts for about 30 per cent of failures. Abdominal salvage for outlet obstruction and low pouch capacitance results in satisfactory or acceptable function in up to 70 per cent of patients. There is no effective surgical salvage for pouchitis. CONCLUSION: Salvage surgery must be discussed carefully with the patient, who should be made aware of the possible complications and the prospect of success, which is less than that in the general population of patients undergoing ileoanal pouch surgery. PMID- 12905543 TI - Options for sphincter preservation in surgery for low rectal cancer. AB - BACKGROUND: Abdominoperineal excision of the rectum with a permanent end-sigmoid colostomy was the classical operation for cancer of the distal third of the rectum. A number of factors have recently led to a more conservative approach, allowing sphincter preservation when excising tumours that are not invading the anal sphincter. METHODS: The review is based on the published literature of the treatment of low rectal cancers accessed by searching Medline and other online databases. It includes a description of all the surgical options currently available for low rectal tumours, and a discussion of the advantages and disadvantages of the types of anastomosis and reconstruction. RESULTS AND CONCLUSION: It is now technically possible to remove rectal cancer that is extending into the anal canal with preservation of the anal sphincter mechanism and with a satisfactory oncological outcome. Ultra-low colorectal and coloanal anastomosis, together with a colonic pouch or coloplasty, produces acceptable function in many patients. However, there is still controversy about the risk of tumour implantation, the place of downsizing neoadjuvant therapy, and true long term functional outcome. Despite these concerns, surgeons should strive to perform rectal resection with sphincter preservation for low-lying rectal cancer whenever possible. PMID- 12905544 TI - Randomized clinical trial of three-layer paste and four-layer bandages for venous leg ulcers. AB - BACKGROUND: Both four-layer and three-layer paste bandages are widely used in the treatment of venous leg ulcers. The aim of this study was to compare the efficacy of these two bandaging regimens. METHODS: The study was a prospective, randomized, open comparison of a consecutive cohort of 133 patients with venous ulcers. Participants were stratified by ulcer size into one of three groups and were randomized within each group to receive either three-layer paste or four layer bandages. All patients were followed for 1 year. The time taken to complete ulcer healing was the primary endpoint. The time taken to apply the bandages, comfort, tolerability and cost were also assessed. Analysis was performed on the basis of intention to treat. RESULTS: Ulcers healed completely in 51 (80 per cent) of 64 patients treated with three-layer paste bandages compared with 45 (65 per cent) of 69 patients treated with the four-layer regimen (P = 0.031). This difference developed only after 20 weeks of treatment. The median times to complete healing were 12 weeks for three-layer and 16 weeks for four-layer treatment (P = 0.040). Results of venous function tests, including half-refilling times, were similar in the two groups. CONCLUSION: Three-layer paste bandages were significantly more effective at healing venous ulcers than the four-layer regimen in this study. PMID- 12905546 TI - Spatial relations of the angular vein, an important landmark in axillary nodal dissection. PMID- 12905545 TI - Randomized clinical trial comparing consultant-led or open access investigation for large bowel symptoms. AB - BACKGROUND: Referral of patients with large bowel symptoms is common and increasing. Currently most of these referrals are assessed at an outpatient clinic to determine the need and priority for investigation. METHODS: Over 21 months, 1131 patients referred by the general practitioner with large bowel symptoms were randomized. Patients in the consultant-led group were assessed by surgeons with a colorectal interest while those in the open access group underwent colonoscopy if they were 55 years or older and flexible sigmoidoscopy if younger. RESULTS: The most common symptom among referred patients was rectal bleeding (69.1 per cent) followed by change in bowel habit (48.8 per cent) and abdominal pain (32.3 per cent). There was a significant trend (P < 0.001) for patients in the consultant-led to have more investigations, and more patients in this group had no investigations (P < 0.001). Despite this, the percentage of patients with colonic or other pathology diagnosed was the same in both groups, 63.6 per cent in the consultant-led group compared with 61.8 per cent in the open access group (P = 0.558). Likewise the percentage of patients with cancer or other significant pathology was similar in both groups (13.9 versus 15.4 per cent; P = 0.532). The mean(s.d.) time to diagnose cancer or other significant pathology was 55.1(39.2) days in the consultant-led group compared with 57.4(33.6) days in the open access group (P = 0.514). The cost per patient was almost pound 105 more for patients in the consultant-led group. CONCLUSION: Patients referred by the general practitioner with large bowel symptoms should go directly to a properly managed and staffed open access large bowel investigation unit. This would enable most patients to have their investigations completed at one hospital attendance. PMID- 12905547 TI - Interstitial laser coagulation with temporary hepatic artery occlusion for patients with cirrhosis and irresectable hepatoma. AB - BACKGROUND: The aim was to determine the degree of local control of hepatocellular carcinoma (HCC) in patients with cirrhotic liver disease when treated with ultrasonographically guided interstitial laser coagulation (ILC) with temporary hepatic artery occlusion. METHODS: Sixteen patients with 24 HCC tumours were treated. Follow-up was by computed tomography or magnetic resonance imaging every 3 months. RESULTS: Nineteen of 24 tumours showed complete necrosis immediately after treatment, and there was no tumour recurrence during follow-up (mean 14 months, median 12 months). No effect on liver function was observed after 1 week and there was no death. In 13 of the 16 patients, new HCC foci developed at other sites. CONCLUSION: Percutaneous ILC combined with temporary hepatic artery occlusion during a single session is an effective local treatment for HCC nodules smaller than 5 cm. However, new HCC lesions develop in the majority of patients, which underscores the need for adjuvant therapy or repeated treatment in these patients. PMID- 12905548 TI - Simultaneous resection of colorectal primary tumour and synchronous liver metastases. AB - BACKGROUND: The surgical strategy for treatment of synchronous colorectal liver metastases remains controversial. The outcome and overall survival of patients presenting with such metastases, treated either by simultaneous resection or by delayed resection, were evaluated. METHODS: From 1987 to 2000, 97 patients presented with synchronous colorectal liver metastases, of whom 35 (36 per cent) underwent a simultaneous resection and 62 patients (64 per cent) a delayed resection. Simultaneous resection was considered prospectively for patients with fewer than four unilobar metastases. RESULTS: Age, blood transfusion requirements, operating time, duration of inflow occlusion, hospital stay and mortality rate were similar in the two groups. The morbidity rate did not differ significantly (23 per cent after simultaneous resection and 32 per cent after delayed resection). The location of the primary tumour and extent of liver resection did not influence the morbidity rate significantly in the simultaneous resection group. The overall survival rate was 94, 45 and 21 per cent at 1, 3 and 5 years respectively after simultaneous resection, and 92, 45 and 22 per cent after delayed resection. CONCLUSION: In selected patients, simultaneous resection of the colorectal primary tumour and liver metastases does not increase mortality or morbidity rates compared with delayed resection, even if a left colectomy and/or a major hepatectomy are required. PMID- 12905550 TI - Laparoscopic surgery for recurrent Crohn's disease. AB - BACKGROUND: The aim of this study was to assess the feasibility of laparoscopic surgery for recurrent Crohn's disease, and the role of repeated laparoscopy in reoperation. METHODS: Between January 1994 and May 2002, 61 laparoscopic operations were attempted in 52 patients with ileal or ileocolonic Crohn's disease. Of these, 16 procedures were performed for recurrence at the anastomotic site (recurrent group). The remaining 45 operations were performed as primary procedures (control group). The median follow-up was 48 (range 3-90) months. RESULTS: The median time to reoperation was 46 months. The incidence of enteric fistula and the conversion rate did not differ significantly between the two groups. Although the operating time was significantly longer in the recurrent group, there were no differences in the rate of postoperative complications (three in the recurrent group and six in the control group) and hospital stay (both median 8 days). CONCLUSION: Laparoscopic surgery for recurrent Crohn's disease is feasible in selected patients without an increase in conversion rate or postoperative complications. PMID- 12905549 TI - Role of neoadjuvant chemotherapy in the treatment of multiple colorectal metastases to the liver. AB - BACKGROUND: The role of neoadjuvant chemotherapy for patients with multiple (five or more) bilobar hepatic metastases irrespective of initial resectability is still under scrutiny. The purpose of this study was to compare the outcome of hepatectomy alone with that of hepatectomy after neoadjuvant chemotherapy for multiple bilobar hepatic metastases from colorectal cancer. METHODS: Retrospective data were collected from 71 patients after hepatectomy for five or more bilobar liver tumours. The outcome of 48 patients treated by neoadjuvant chemotherapy followed by hepatectomy was compared with that of 23 patients treated by hepatectomy alone. RESULTS: Patients who received neoadjuvant chemotherapy had better 3- and 5-year survival rates from the time of diagnosis than those who did not (67.0 and 38.9 versus 51.8 and 20.7 per cent respectively; P = 0.039), and required fewer extended hepatectomies (four segments or more) (39 of 48 versus 23 of 23; P = 0.027). Multivariate analysis showed neoadjuvant chemotherapy to be an independent predictor of survival. CONCLUSION: In patients with bilateral multiple colorectal liver metastases, neoadjuvant chemotherapy before hepatectomy was associated with improved survival and enabled complete resection with fewer extended hepatectomies. PMID- 12905551 TI - Recurrence and survival after mesorectal excision for rectal cancer. AB - BACKGROUND: Mesorectal excision for rectal cancer has resulted in local recurrence rates of 3-11 per cent compared with up to 38 per cent after conventional methods. The results of a prospective Danish study with a historical control group are presented. METHODS: Three hundred and eleven patients with a mobile rectal cancer had mesorectal excision with curative intent performed by certified surgeons and were followed for 3 years. Demographic, perioperative and follow-up data were recorded prospectively. A series of patients who had conventional operations for rectal cancer served as a control group. RESULTS: The cumulative 3-year local recurrence rate was 11 per cent after mesorectal excision compared with 30 per cent after conventional surgery (hazard ratio (HR) 0.33 (95 per cent confidence interval (c.i.) 0.21 to 0.52); P < 0.001). Multivariate regression analysis showed that only advanced age (HR 0.97 (95 per cent c.i. 0.94 to 1.00); P = 0.048) and tumour in the lower third of the rectum (HR 0.21 (95 per cent c.i. 0.04 to 1.97); P = 0.075) were marginal independent predictors of local recurrence after mesorectal excision. The cumulative crude 3-year survival rate was 77 per cent after mesorectal excision and 62 per cent after conventional surgery (HR 0.58 (95 per cent c.i. 0.43 to 0.77); P < 0.001). Age was the only independent predictor of death after mesorectal excision (HR 1.04 (95 per cent c.i. 1.02 to 1.07); P = 0.001). CONCLUSION: Mesorectal excision is associated with a considerably lower risk of local recurrence and a better survival rate than conventional surgery, and is the optimum method for rectal cancer resection. PMID- 12905552 TI - Colonic anastomotic strength and matrix metalloproteinase activity in an experimental model of bacterial peritonitis. AB - BACKGROUND: Clinical studies report conflicting results on the safety of primary intestinal anastomoses in the presence of peritonitis, and comprehensive experimental data are lacking. The present study investigated whether the strength of experimental colonic anastomoses is affected if surgery is performed in the presence of pre-existing bacterial peritonitis. METHODS: Colonic anastomoses were constructed in Wistar rats 24 h after caecal ligation and puncture or a sham procedure. Anastomotic strength was assessed by measuring breaking strength and bursting pressure during the first 5 days after operation. Anastomotic hydroxyproline levels were measured and matrix metalloproteinase (MMP) activity was analysed by quantitative gelatin zymography. RESULTS: Anastomotic strength was lowered in the presence of bacterial peritonitis but in a minor and transient way. The breaking strength was lower only immediately after construction of the anastomosis (- 15 per cent, P = 0.011) and the bursting pressure only on the third postoperative day (- 33 per cent, P = 0.038); no anastomotic dehiscence was observed. At 3 days after operation increased levels of MMP activity were observed but anastomotic hydroxyproline content was not affected by bacterial peritonitis. CONCLUSION: The influence of bacterial peritonitis on the development of anastomotic strength is limited. This experimental finding lends support to recent clinical studies that have demonstrated the feasibility of constructing a primary anastomosis under these conditions. PMID- 12905553 TI - Detection of disseminated tumour cells in blood and bone marrow samples of patients undergoing hepatic resection for metastasis of colorectal cancer. AB - BACKGROUND: In 50-60 per cent of patients who undergo hepatic resection for metastasis of colorectal cancer the first site of tumour recurrence is extrahepatic, indicating the presence of more extensive disease at the time of resection. The aim of this study was to evaluate whether the presence of disseminated tumour cells in blood and bone marrow could predict extrahepatic tumour recurrence. METHODS: Cytokeratin 20 (CK20) reverse transcriptase polymerase chain reaction was used to study the presence of tumour cells in preoperative peripheral blood and bone marrow samples from 41 patients with liver metastasis scheduled for surgical resection. RESULTS: CK20 expression was detected in six of 41 peripheral blood samples and in eight of 32 bone marrow samples. There was no correlation between CK20-positive samples and subsequent extrahepatic recurrence. Positive blood samples did, however, correlate with high serum carcinoembryonic antigen level and large tumour volume. None of the 14 patients previously treated with chemotherapy had CK20-positive samples, whereas six of 27 blood and eight of 20 bone marrow samples were positive in the chemotherapy-naive group. CONCLUSION: Although the number of patients in this study is limited, the presence of disseminated tumour cells did not predict subsequent extrahepatic recurrence. The results strongly suggest that the presence of circulating tumour cells in peripheral blood may reflect transient shedding of tumour cells related to large tumour volume. PMID- 12905554 TI - Long-term results of the Lyons R90-01 randomized trial of preoperative radiotherapy with delayed surgery and its effect on sphincter-saving surgery in rectal cancer. PMID- 12905555 TI - Optimal preoperative assessment and surgery for rectal cancer may greatly limit the need for radiotherapy. AB - BACKGROUND: Radiation is being used increasingly in the management of patients with rectal cancer. Over the past decade the Basingstoke Colorectal Research Unit has combined precision total mesorectal excision with the highly selective use of preoperative radiotherapy. METHODS: One hundred and fifty consecutive patients who underwent major surgical excision for cancers of all stages comprised the study group. Preoperative clinical assessment was based largely on tumour size, fixation and distance from the anal verge. Only preoperative radiotherapy was considered and this only for tumours judged to be at high risk of mesorectal fascia involvement. RESULTS: During a 5-year period 35 of 150 patients were selected for preoperative irradiation. In the non-irradiated patients the local recurrence rate after a median follow-up period of 870 (range 51-1903) days was 2.6 per cent (three of 115 patients), compared with 17.1 per cent (six of 35 patients) in those chosen for irradiation. Sixty patients (52.2 per cent) who were not irradiated were node positive. The local recurrence rate for the whole group was 6.0 per cent. CONCLUSION: The great majority of patients undergoing major excision for rectal cancer can be managed without radiation therapy if the preoperative assessment of the mesorectal fascia and surgery are performed optimally. PMID- 12905556 TI - Influence of suture material and surgical technique on risk of reoperation after non-mesh open hernia repair. AB - BACKGROUND: Although mesh techniques are used with increasing frequency, sutured repair still has a place in groin hernia surgery. Studies relating suture material to recurrence rate have yielded conflicting results. The aim of the present study was to analyse the influence of suture material and sutured non mesh technique on the risk of reoperation in open groin hernia repair using data from the Swedish Hernia Register. METHODS: The relative risk of reoperation after sutured repair using non-absorbable, late absorbable and early absorbable sutures was compared in multivariate analyses, taking into account known confounding factors. RESULTS: Between 1992 and 2000, 46,745 hernia repairs were recorded in the Swedish Hernia Register. Of these, 18,057 repairs were performed with open non-mesh methods and were included in the analysis. Using non-absorbable suture as reference, the relative risk of reoperation after repair with early absorbable suture and late absorbable suture was 1.50 (95 per cent confidence interval (c.i.) 1.22 to 1.83) and 1.03 (95 per cent c.i. 0.83 to 1.28) respectively. Using the Shouldice repair as reference, other sutured repairs were associated with a significantly higher relative risk of reoperation (1.22, 95 per cent c.i. 1.03 to 1.44). CONCLUSION: A non-absorbable or a late absorbable suture is recommended for open non-mesh groin hernia repair. The Shouldice technique was found to be superior to other open methods. PMID- 12905558 TI - Protection against experimental small intestinal ischaemia-reperfusion injury with oxygenated perfluorochemical. AB - BACKGROUND: Intestinal ischaemia-reperfusion (IR) injury frequently occurs in abdominal surgery. Perfluorochemical (PFC) can be used to oxygenate intestinal organs directly and allows adenosine 5'-triphosphate (ATP) production within the submerged organs during ischaemia. This study was designed to evaluate the protective effect of PFC in IR injury, focusing on cytokine production in rat small intestine. METHODS: The superior mesenteric artery was occluded in rats for 60 min and the small bowel placed in an intestinal bag containing either normal saline (group 1), oxygenated saline (group 2) or oxygenated PFC (group 3). The arterial clip was subsequently removed, allowing reperfusion. The number of rats that survived for 7 days, tissue ATP levels, biochemical variables, tissue lipid peroxidation (LPO), bacterial cultures and histological changes were examined after reperfusion. RESULTS: The use of oxygenated PFC in group 3 improved survival compared with the other groups. Serum creatine phosphokinase and lactate dehydrogenase levels in groups 1 and 2 reflected small intestinal damage, and plasma levels of tumour necrosis factor alpha and interleukin 6 were raised. In contrast, oxygenated PFC decreased these levels, and reduced LPO, bacterial translocation and augmented apoptosis of the small intestine after reperfusion. CONCLUSION: An intestinal bag containing oxygenated PFC showed protective effects during bowel ischaemia. PMID- 12905557 TI - Fibrosis-associated gene expression in renal transplant glomeruli after acute renal allograft rejection. AB - BACKGROUND: Acute allograft rejection is thought to be a risk factor for chronic allograft nephropathy, the cardinal features of which are vasculopathy, interstitial fibrosis and glomerulosclerosis. Fibrosis-associated genes might act as ad interim surrogate markers for chronic allograft nephropathy. The aim of this study was to determine mRNA expression of fibrosis-associated genes in glomeruli plucked from protocol renal transplant biopsies, in patients with or without a history of acute rejection. METHODS: A consecutive series of 52 patients (31 male, 21 female) was assessed. Donor categories were cadaveric, living related or asystolic. Transplant recipients received either cyclosporin- or tacrolimus-based immunosuppression. Patients routinely underwent percutaneous needle-core renal transplant biopsy at 1 week, and 3 and 6 months. Acute rejection episodes were confirmed histologically and treated with intravenous methylprednisolone, or antithymocyte globulin if steroid resistant. Individual glomeruli were plucked and total mRNA was extracted. Fibrosis-associated genes were amplified by reverse transcriptase-polymerase chain reaction (PCR) and quantified by enzyme-linked immunosorbent assay. RESULTS: The expression of both collagen type III (mean 0.42 versus 0.31 arbitrary units of PCR products corrected for a housekeeping gene) and collagen IV (mean 0.46 versus 0.42 arbitrary units) at 6 months did not differ between recipients who experienced acute rejection episodes and those who were free from rejection. There was also no significant difference between groups in terms of mRNA expression of collagen IValpha2, matrix metalloproteinase 2, tissue inhibitor of matrix metalloproteinases 1 and 2, transforming growth factor beta or tenascin. CONCLUSION: These results suggest that acute rejection episodes do not increase the expression of fibrosis-associated genes in glomeruli from renal transplant biopsies. PMID- 12905559 TI - Evaluation of POSSUM and P-POSSUM scoring systems in patients undergoing colorectal surgery (Br J Surg 2003; 90: 340-345). PMID- 12905561 TI - Temporary closure of open abdominal wounds by the modified sandwich-vacuum pack technique (Br J Surg 2003; 90: 718-722). PMID- 12905562 TI - Randomized clinical study of Gastrografin administration in patients with adhesive small bowel obstruction (Br J Surg 2003; 90: 542-546). PMID- 12905563 TI - Factors predictive of 5-year survival after transarterial chemoembolization for inoperable hepatocellular carcinoma (Br J Surg 2003; 90: 325-331). PMID- 12905566 TI - A tribute to Daniel Perry. Interview by Val J. Halamandaris. PMID- 12905567 TI - Total turnaround: how to use national benchmarks, an operational review, and focus change strategies to totally turn an agency around. PMID- 12905568 TI - How to maximize the return on your OASIS investment. PMID- 12905569 TI - Metabolic syndrome & obesity: co-epidemics could overwhelm home health care. AB - Our nation's battle with the bulge has contributed to the current co-epidemics of obesity and metabolic syndrome. The US Surgeon General's office reports that these disease risks may soon cause as much disease and death as cigarette smoking (HHS, 2001). In the United States, one in four adults has metabolic syndrome, which includes about 15 million persons with type 2 diabetes (Ford, 2002). About 50 percent of the estimated 47 million individuals with metabolic syndrome also have insulin resistance, which is a major factor in type 2 diabetes (Ford, 2002; Blackburn, 2001). Overweight and obese adults have a 50 to 100 percent increased risk for premature death compared to those of normal weight (HHS, 2001). The co epidemics of obesity and metabolic syndrome are costly, as well, having an estimated price tag in 2000 of over $117 billion (HHS, 2001). This article will define the co-epidemics of obesity and metabolic syndrome and describe recommended prevention and management procedures to be adopted in home care. PMID- 12905570 TI - Knowing where you're going changes the journey. PMID- 12905571 TI - [Experiment study of c-fos expression on myocardial acute ischemia/reperfusion injury in rats]. AB - OBJECTIVE: To investigate the changes of c-fos mRNA induced by myocardial ischemia/reperfusion(I/R) during acute period. METHODS: The model of I/R was established, and the rats was divided normal control, ischmia and I/R groups. Insitu hybridization (ISH) and computerized image analysis method was used to observe alteration of c-fos mRNA in cardiac myocytes. RESULTS: After 10 min ischemia and 30 min reperfusion, the area of reperfusion showed a part of cardiac myocytes weak staining, and get to peak at I-60 min/R-30 min. The myocardium in normal control and ischemia groups less than 60 min showed negative staining. No changes were found in all groups by HE staining. CONCLUSION: c-fos mRNA detection may become an important method for diagnosis of I/R. PMID- 12905572 TI - [Study on the changes of ncNOS in chronic heroin dependence and spontaneous withdrawal in rats]. AB - OBJECTIVE: To study the changes and actions of neuronal constructive nitric oxide synthase(ncNOS) in heroin drug abuse. METHODS: The expression of ncNOS and ncNOS mRNA in neurons of cerebral cortex, periaqueductal gray matter and the ventral tegmental area was observed by immunohistochemistry, in situ hybridization and image analysis technique after heroin dependence and spontaneous withdrawal in rats. RESULTS: The quantity of ncNOS and ncNOS mRNA rised clearly and the number of ncNOS and ncNOS mRNA positive cells increased greatly in heroin dependence and withdrawal. The changes of ncNOS and ncNOS mRNA in spontaneous withdrawal were more clear than ones of dependence. Heroin dependence and withdrawal led to alterations in ncNOS and ncNOS mRNA expression in important regions implicated in the physical tolerance and dependence. CONCLUSION: The ncNOS plays an important role in heroin dependence and withdrawal. The ncNOS immunohistochemical changes observed in the present study might be useful for the forensic pathological diagnosis of heroin drug abuse. PMID- 12905573 TI - [The development and changes of pressure of putrefactive gas in cadaveric enterocelia in spring and its forensic application]. AB - OBJECTIVE: To study the development and changes of pressure of putrefactive gas (PPG) in cadaveric enterocelia in spring, and to explore its application in estimation of postmortem interval (EPI). METHODS: 57 goats were divided into 2 groups according to means of death, on land or in water. Celiac PPG were observed timely and systematically. RESULTS: The development of PPG in cadaveric enterocelia, which can be divided into raising phase, peak phase, and declining phase was observed, and a model to estimate postmortern interval by changes of PPG was founded. CONCLUSION: Measuring PPG in cadaveric enterocelia could be used in forensic EPI. PMID- 12905574 TI - [The relationship between the postmortem interval and the changes of DNA content in rabbit's cornea epithelial and endothelium by the computerized image analysis]. AB - OBJECTIVE: To study the relationship between the postmortem interval (PMI) and the changes of DNA content in rabbit's cornea epithelial and endothelium. METHODS: Using the computerized image analysis to measure the DNA content in the cornea epithelial and endothelium of 105 rabbit's at every six hours during 72 hours after death. RESULTS: The degradation rate of DNA in nuclear has an apparent relationship in 72 hours after death of the rabbits. CONCLUSION: The determination of the quantity of DNA in cornea nuclear is a new method to estimate the PMI accurately. PMID- 12905575 TI - [A study on the relationship between the expression of apoptosis-related gene and the injury time in earlier period following cerebral contusion]. AB - OBJECTIVE: To investigate the relationship between the expressive variety of apoptosis-related gene Bcl-2, Fas and the brain injury time during earlier period after cerebral contusion. METHODS: The morphologia feature of brain tissue was observed under microscopy and immunohistochemistry, image analyze technique was applied to detected the expression of Bcl-2 and Fas. RESULTS: Typical apoptotic cells were seen 8 hours after cerebral contusion. The expression of apoptosis related gene was noted after 30 minutes and keep increase subsequently. The expression of Bcl-2 reached maximum level at 4 hours after brain trauma and then declined. CONCLUSION: Detecting the expression of Bcl-2 and Fas would be of valuable in the distinguishment of promortem and postmortem injury, and the estimation of brain injury time during the earlier period after cerebral contusion. PMID- 12905577 TI - [Age determination by radiographic feature of thyroid cartilage in female Han population]. AB - OBJECTIVE: To seek a method of age determination. METHODS: This sample consists of 69 female Chinese han individuals whose ages been known. Five positions were selected from their radiograms which have been divided into three or six grade and then set up a mathematical model to deduce age by multiple linear regression analysis. RESULTS: The standard deviation of female thyroid cartilage is 1.969 and the multiple correctation coefficient is 0.984. CONCLUSION: The observation proved that age determination by radiographic feature of thyroid cartilage can be used for female Chinese han population from 18 to 60 years old. PMID- 12905576 TI - [Experimental pathological study of subacute intoxication by Dioscorea bulbifera L]. AB - OBJECTIVE: To study the pathological change and the toxic mechanism of Dioscorea bulbifera L in mice. METHODS: Sixty ICR mice were randomly assigned to four groups poisoned respectively with 200% Dioscorea bulbifera L of 1/4 LD50, 1/10LD50, 1/30LD50 and a control group treated with distilled water by oral administration. All animals were pathologically examined with LM and some of them were examined with TEM when the mice died during the experiment or the survival mice were sacrificed after thirty days. RESULTS: The pathological changes showed fatty change and the increasing glycogen of liver cells; degeneration and necrosis of the epithelia of uriniferous tubules. The serum BUN and ALT of the experimental groups mice were higher than that of control group. Enzyme histochemical staining showed the decreasing activity of G-6-P and SDH in the liver cells in the experimental groups. CONCLUSION: The experiment suggests that the target organs were liver and kidney. The toxic mechanism of Dioscorea bublifera L was the damage of the mitochondrional and endoplasmic reticulum membrane directly. As a result, the activity of the SDH and G-6-P decreased, the metabolism was affected. PMID- 12905579 TI - [Factors affecting the determination of the percent carboxyhemoglobin saturation of blood]. AB - OBJECTIVE: To investigate factors affecting the determination of the percent carboxyhemoglobin saturation (HbCO%) of blood in an attempt to offer further data for results interpretation and sample storage requirement. METHODS: The HbCO% of blood samples stored in various conditions were detected by three spectrophotometries during the succeeding 30 days. RESULTS: The data detected by reductive double-wavelength spectrophotometry and double-wavelength spectrophotometry were more stable than mono-wavelength spectrophotometry. The HbCO% of blood was significantly related with the storage conditions which include temperature, time and the degree of exposure to air. CONCLUSION: Determinations of HbCO% are reliable which performed by reductive double wavelength spectrophotometry and double-wavelength spectrophotometry, combine with spectral scans. During 30 days, blood stored at 4 degrees C exposed to a limited volume of air does not influence the determination of HbCO%. PMID- 12905578 TI - [A study on sarcosaphagous insects species variety with seasons in Chengdu]. AB - OBJECTIVE: To study the species of sarcosaphagous insects. METHODS: Rabbits were killed and placed outdoors from March to November. Flies that appeared the cadavers were observed and identified. RESULTS: There are five main flies, i.e., Muscadomesticauicina, Lucilia sericata, C. Megacephala, S. fuscicauda, Aldrichiragrahormi. From midtime of April to the beginning of October, Muscadomesticauicina could be seen on the cadvers, however, Aldrichiragrahormi only could be seen before the beginning of May. Otherwise, Lucilia sericata, C. Megacephala, S. fuscicauda could be seen on the cadvers from March to the end of October. After the midtime of November, none of sarcosaphagous flies could be seen on the cadvers. On the other hand, maggots of sarcosaphagous flies could be seen often on the cadvers after adults of sarcosaphagous flies intruding 1 to 4 days, relating to temperature of environment. CONCLUSION: If these flies history of life be studied. it is useful for estimating postmortem interval in Chengdu. PMID- 12905580 TI - [Estimation of postmortal interval by measuring the amount of DNA in cells]. AB - Although there are many methods to estimate the early postmortal interval, more attention has been paid to the research on measuring the amount of DNA in cells. This paper introduce several different measuring ways, law of variation and application situation of DNA in cells. In addition, the result evaluation of measuring methods and application prospect is given. PMID- 12905581 TI - [The role of macrophages in wound age estimation during the wound healing process]. AB - The article reviewed the advance and the forensic meaning of macrophage on the wound age estimation during the wound healing process in resent years. It has also been summarized the relationship between macrophages and wound age estimation on the expression of cytokines derived from macrophages, the changes of macrophage phenotypes and the development process of phagosomes in macrophages after wounding. It is suggested that some regular and characteristic changes of macrophage should be a useful mark to wound age estimation and reminds therefore further study in this field. PMID- 12905582 TI - [The research of the heroin and its metabolites analysis in clinical samples]. AB - Heroin can be metabolized easily in body and the mail metabolites are 6-MAM, morphine and so on. At present, there are urine, blood, hair and so on as specimens for detection, while the analytical technology conclude TLC, GC, HPLC, GC/MS, LC/MS, IA, CE etc. In this paper, these technologies used for heroin's metabolites were viewed in order to provide some reference to the study in relative field. PMID- 12905583 TI - Individual account retirement plans: an analysis of the 2001 Survey of Consumer Finances. PMID- 12905584 TI - Emergency departments. Smoothing the flow. PMID- 12905585 TI - Military hospitals. Wired at war. PMID- 12905586 TI - Workforce. Body politic. PMID- 12905587 TI - Readiness. Three days in May. PMID- 12905588 TI - The hand that feeds you. PMID- 12905589 TI - Management. CMOs' role expands. PMID- 12905590 TI - Community outreach. Give me the news. PMID- 12905591 TI - The data page. Heavy user. PMID- 12905592 TI - Wilson's pianos. PMID- 12905593 TI - Most wired 2003. AB - A record number of health care organizations are represented in H&HN's fifth Most Wired Survey and Benchmarking Study. This year's survey reveals numerous trends in IT strategies among the 100 Most Wired, and the winners of the Innovator, Most Improved and Most Wireless awards. New this year: the Most Wired Small and Rural Award. PMID- 12905594 TI - Global health report. A snapshot of the payer systems, major diseases and workforce trends from around the world. AB - International trade and travel have rendered political and geographic boundaries irrelevant when it comes to diseases. HIV and SARS are among the more recent examples of deadly viruses that spread swiftly from one continent to the next before international health experts can identify them, much less begin to search for a cure. This snapshot of global health issues looks at payer systems from a cross section of nations, lists the world's eight most infectious diseases and their rate of incidence by region, and pays special attention to the shortage of health care workers, which has reached crisis proportions in many areas and is pitting nation against nation in the competition for qualified staff. PMID- 12905595 TI - 2003 NOVA award winners. AB - Even in tough economic times, the five recipients of this year's NOVA Award understand the value of community outreach. Improving the health and well-being of the people they serve is a cornerstone of their mission and it means going outside the walls of their institutions. They also understand that getting the most out of their ambitious community programs requires cooperation with other groups. "If you just push a little more in the face of challenging financial times, through collaboration, everyone benefits," says Gregory Wozniak, president and CEO of NOVA winner St. Mary Medical Center, Langhorne, Pa. "That's one way to decrease costs and increase the access of health care." PMID- 12905596 TI - Honest talk. Disclosure is not about asset protection--it is about doing the right thing. PMID- 12905597 TI - [Ectopic osteogenesis of tissue engineered bone with slow-released bioactive factors]. AB - OBJECTIVE: To study the ectopic osteogenesis of tissue engineered bone with recombinant human bone morphogenetic protein/transforming growth factor-beta (rhBMP/TGF-beta) or WO-1 slow-released factors. METHODS: Partial demineralized freeze-dried bone (PDFDB) of pig was used as scaffold material. rhBMP/TGF-beta or WO-1 were pre-coated on the surface of material by means of vacuum negative pressure absorption, and then coated with polylactic acid (PLA) to make slow released material. There were six group: PDFDB material (group A); PDFDB combined with osteoblasts (group B); PDFDB material with rhBMP/TGF-beta slow-released system (group C); PDFDB material combined with rhBMP/TGF-beta slow-released system osteoblasts (group D); PDFDB with WO-1 slow-released system (group E); PDFDB material combined with WO-1 slow-released system and osteoblasts (group F) were implanted in bilateral lower limbs of 36 Newzealand rabbits respectively (6 rabbits in each groups). Histological, histochemical and biochemical analysis were detected 2, 4, 6, 8 weeks after operation. RESULTS: Within the observation periods, no osteogenesis was observed in group A. The osteogenesis in group B, D, F were superior to that of group C and E (P < 0.05). The osteogenetic activity in group C and E was delayed. The quantity and quality of osteogenesis in group D and F were 2 weeks ahead of time compared with group B, and 4 weeks to that of group C and E. The newborn calcification content was superior to that of group A, C, and E (P < 0.05). CONCLUSIONS: The osteogenesis of PDFDB materials with BMP/TGF-beta or WO-1 is slower than that of which combined with osteoblasts. Simple material PDFDB has no ectopic osteogenesis. PMID- 12905598 TI - [Experimental study of cardiac muscle tissue engineering in bioreactor]. AB - OBJECTIVE: This study investigates construction of cardiac muscle cell-porous collagen scaffold complex in a bioreactor so as to unveil the possibility of generating 3-dimensional cardiac muscle tissue under the environment that mimics microgravity in vitro. METHODS: 1-2-day old neonatal rat cardiac muscle cells were isolated by sequential digestion and pre-plating methods, then seeded onto porous collagen scaffold. The cell-collagen complex was transferred into rotary cell culture system (RCCS) and incubated for 7 days. Cells cultured in 75 ml flasks and constructs cultures on plates served as control. Morphological changes of the cells were observed by light microscope and metabolic rate was recorded. Ultrastructure of the cells growing in porous collagen was observed by transmission electron microscopy. Content of total DNA and protein in the newly formed tissue were analyzed. H-E and anti-sarcomeric alpha-actin stains were performed in comparison with native cardiac muscle. RESULTS: The isolated cardiac muscle cells adhered to the bottom of the flasks 24 hours after plating and began to beat spontaneously. When incubated for 7 days in RCCS, cell-collagen constructs of form a continuous outer tissue layer containing cells aligned with each other. The cell population in the interior of the construct was less in density than the outer part. Transmission electron microscopy demonstrated that subcellular elements characteristic of cardiac myocytes were in the outermost layer of constructs. A strongly positive stains of anti-sarcomeric alpha-actin suggested presence of cell population of differentiated cardiac myocytes in these constructs. Construct biomass was not significantly different from that in neonatal rat ventricle and approximately 40% of that in adult rat ventricles. Construsts in plates contained a few of cells which were less than those in RCCS. Metabolic activity of cells cultured in RCCS was higher than that in flasks and plates. CONCLUSIONS: Dissociated cardiac muscle cells cultured on 3-dimensional scaffolds in RCCS under favorable conditions can form engineered constructs with structural and functional features resembling those of native cardiac tissue. PMID- 12905599 TI - [Effects of transient transfection of human BMP-4 gene on rabbit bone marrow stromal cells]. AB - OBJECTIVE: Bone marrow stromal cells (MSCs) were transfected with human bone morphogenetic protein-4 (hBMP-4) gene in vitro to provide BMP gene modified cells for tissue-engineered bone. METHODS: MSCs were cultured and transfected with pEGFP-hBMP4, pEGFP plasmids respectively or left uninfected as control. Transcription of BMP-4 gene as well as gene transfection efficiency was tested. Morphological and growth feature of the transfected cells were valued. Alkaline phosphatase (ALP), von Kossa, and Osteocalcin (OC) were tested to determine the phenotypes of osteoblast. RESULTS: The gene transfection efficiency was 20%-30%, based on GFP expression. RT-PCR showed that MSCs had low transcription of BMP-4 that was enhanced by the gene transfer. Morphological feature of MSCs transfected with pEGFP-hBMP-4 changed but growth curves did not show much difference among the groups. In pEGFP-hBMP-4 group, ALP positive stain area and the number of calcium nodules were increased, as well as the expression of OC. CONCLUSIONS: A high transfer efficiency of MSCs was achieved under optimized conditions. The gene transfer technique strengthened the transcription of BMP-4 and promoted differentiation from MSCs to osteoblasts. hBMP-4 transferred MSCs may serve as an ideal cell source for tissue-engineered bone. PMID- 12905600 TI - [The construction of two kinds of artificial bone and the comparison of their osteogenesis capability in vivo]. AB - OBJECTIVE: To construct artificial bone with collagen-hydroxyapatite (HA) or collagen-HA-chondroitin sulfate (CS) as the scaffolds, and observe their biological properties. METHODS: The artificial bones were constructed by attaching recombinant human bone morphogenetic protein 2 (rhBMP-2) on those scaffolds. And then they were embedded into muscles of rats. Every weekend those newly formed bones were taken from muscles for comparing the difference in osteogenetic capability of two kinds of artificial bone in vivo. RESULTS: Both kinds of artificial bones could induce bone regeneration in muscle. The collagen HA-CS artificial bone was superior to the collagen-HA artificial in bone-guided degree. CONCLUSIONS: The CS could promote the form action of new bone and accelerate the bone healing. PMID- 12905601 TI - [The regulatory effect of human bone morphogenetic protein 7 gene transfer on the proliferation and differentiation of rabbit bone marrow mesenchymal stem cells]. AB - OBJECTIVE: To detect the proliferation and differentiation of rabbit bone marrow mesenchymal stem cells (BMSc) transferred by retroviral vector carrying human bone morphogenetic protein 7 (hBMP-7) gene. METHODS: hBMP-7-expressing replication-deficient retroviral vector(PT-PLNCX2-hBMP7) was reconstructed using clone technique and recombinant DNA technique. BMSc were infected with the virus granules. The protein of BMP-7 gene in transferred cells were determined by immunohistochemistry. The proliferativity of the transferred cell were assayed by methabenzthiazuron (MTT) method and flow cytometer. Alkaline phosphatase (ALP) were also detected using enzyme kinetics. RESULTS: Cells transferred by PT-PLNCX2 hBMP7 expressed abundant hBMP7 protein in the cytoplasm. Positive findings were not found in those cells that were not transferred. After infected with virus there were not significant difference of cell proliferation and cell cycle between the cells transferred by hBMP-7 or not (P > 0.05). ALP activity in transferred cells were increased significantly (P < 0.01). CONCLUSIONS: hBMP-7 can be transferred and stably expressed in the cultured rabbit bone marrow stem cells. Proliferation and cell cycle of the transferred cell were not affected. hBMP7 gene transfer can be used to induce differentiation of BMSc into osteoblast like cells. PMID- 12905602 TI - [The method of accelerating osteanagenesis and revascularization of tissue engineered bone in big animal in vivo]. AB - OBJECTIVE: To study whether tissue engineered bone can repair the large segment bone defect of large animal or not. To observe what character the fascia flap played during the osteanagenesis and revascularization process of tissue engineered bone. METHODS: 9 Chinese goats were made 2 cm left tibia diaphyseal defect. The repairing effect of the defects was evaluated by ECT, X-ray and histology. 27 goats were divided into three groups: group of CHAP, the defect was filled with coral hydroxyapatite (CHAP); group of tissue engineered bone, the defect was filled with CHAP + bone marrow stroma cells (BMSc); group of fascia flap, the defect was filled with CHAP + BMSc + fascia flap. After finished culturing and inducing the BMSc, CHAP of group of tissue engineered bone and of fascia flap was combined with it. Making fascia flap, different materials as described above were then implanted separately into the defects. Radionuclide bone imaging was used to monitor the revascularization of the implants at 2, 4, 8 weeks after operation. X-ray examination, optical density index of X-ray film, V G staining of tissue slice of the implants were used at 4, 8, 12 weeks after operation, and the biomechanical character of the specimens were tested at 12 weeks post operation. RESULTS: In the first study, the defect showed no bone regeneration phenomenon. 2 cm tibia defect was an ideal animal model. In the second study, group of CHAP manifested a little trace of bone regeneration, as to group of tissue engineered bone, the defect was almost repaired totally. In group of fascia flap, with the assistance of fascia flap which gave more chance to making implants to get more nutrient, the repair was quite complete. CONCLUSIONS: The model of 2 cm caprine tibia diaphyseal defect cannot be repaired by goat itself and can satisfy the tissue engineering's demands. Tissue engineered bone had good ability to repair large segment tibia defect of goat. Fascia flap can accelerate the revascularization process of tissue engineered bone. And by this way, it augment the ability of tissue engineered bone to repair the large bone defect of goat. PMID- 12905603 TI - [An experimental study of demineralized bone matrix to repair bone defects as a scaffold of tissue engineering]. AB - OBJECTIVE: To evaluate application of the sponge of demineralized bone matrix (SDBM) in tissue engineering of bone. METHODS: SDBM was prepared from long bone of rabbits. Bone marrow cells were flushed from the bone shaft of femurs of a two month-old New Zealand white rabbit. After the cells were cultured for 9 days, the flasks were added into dexamethasone (10(-8) mol/L), beta-glycerophosphate sodium (10 mmol/L) and L-ascorbic acid (50 micrograms/ml). After 5 weeks, the cultured cells were collected and marked by 5-Bromo-2'-dexyouridine (BrdU). The grand sum of cells seeded on a piece of SDBM was about (4-6) x 10(6). The composites of cells and SDBM (tissue engineered chip, TEC) were implanted into muscles and bone defects of radius in rabbits. A standard procedure was applied to make a 10 mm long defect bilaterally in the radius of nine skeletally mature male New Zealand white rabbits. All of the 18 defects were randomly divided into three groups: group I, six defects were grafted by TEC; group II, six defects were grafted with SDBM alone; group III, six defects were empty. RESULTS: The results of radiographic and histological evaluation showed that all of the defects were repaired in group I and group II at 6 weeks, none of the defects was repaired in group III. The results of BrdU staining showed that the staining was positive in group I, but negative in group II. Biomechanical test showed that the compressive ultimate strength (CUS) of new bone in TEC implanted group was comparable with normal radius (P = 0.623) and in SDBM implanted group was significant lower than normal radius (P = 0.038). CONCLUSIONS: The TEC can form cartilage and bone tissue in muscles and repair segmental bone defects. SDBM is a kind of effective natural scaffold in tissue engineering of bone. PMID- 12905604 TI - [Mineralized collagen based composite for bone tissue engineering]. AB - OBJECTIVE: To construct a mineralized collagen based composite by biomimetic synthesis for bone tissue engineering. METHODS: Using the molecular collagen as the template, the calcium phosphate is deposited on it to produce a mineralized collagen based composite, then is combined with minute amount of poly lactic acid (PLA), the three-dimensional scaffold composite is prepared by liquid phase separation. Using osteoblast culture technique, the biocompatibility of this biomaterial in vitro is detected by x-ray diffraction, SEM, TEM, fluoroscopy and CLSM. RESULTS: Both degree and the size of crystals in the composite are low, which are similar to that of nature bone. It possesses porous structure and the porosity of the composite is high. The typical fibrillar microstructure is self assembled of the collagen and the nano-crystal hydroxyapatite (HA) in the composite, moreover, the x-ray diffraction graphic of HA crystal shows the [002] oriented. CONCLUSIONS: The biomimetic three-dimensional composite can serve as one of the optimal scaffold material for bone tissue engineering both on structure and on property. PMID- 12905605 TI - [Experimental research on spine fusion induced by tissue engineered bone]. AB - OBJECTIVE: To construct a reasonable substitute for the autograft bone in vitro and transplant it back into the rabbit models to induce the spine fusion. METHODS: The bone marrow stem cell from the seven New Zealand rabbits were cultured. Recombinant human bone morphogenetic protein-4 (rhBMP-4) that has been proved to be bioactive was obtained by the way of genetic engineering. Using the vacuum freezing machine to mix a certain quantity of rhBMP-4 into type I collagen to form a new kind of carrier. Animal model of spine facet process fusion was used. Bone marrow stem cells combined with rhBMP-4 and I type collagen were implanted between the facet process to induce the spine fusion. type I collagen and bone marrow stem cell was used in the controlled group. RESULTS: New bone formation was obvious in the test group. The facet joint was fused very well in this side. No bone formation was present on the other side. CONCLUSIONS: The new composite: bone marrow stem cells, rhBMP-4 and type I collagen was an ideal kind of substitute for the autograft bone. PMID- 12905606 TI - [Experimental study of the immuno-protective activity of recombinant vaccinia virus expressing HPV58 E7]. AB - OBJECTIVE: To develop a recombinant vaccinia virus vaccine expressing HPV58 E7 and to determine its immuno-protective activity in mice bearing HPV58 E7+ tumor. METHODS: E7 DNA was amplified and cloned from a plasmid containing HPV58 E7 genome by PCR. To abolish its transforming activity, the nucleotides coding for amino acid residues at positions 24 and 92 were modified by site-directed mutagenesis so that cysteine was substituted by glycine. Balb/c 3T3 cells were transfected with mE7. The expression of E7 protein by the mE7-transfected Balb/c cells was confirmed by immunofluorescence staining. The transfected cells were observed in vitro for anchorage-independent growth and tumorigenesis in nude mice. Recombinant E7 vaccinia virus vaccine was constructed by homologous recombination of HPV58 E7 vaccinia expression plasmid and vaccinia virus (Tiantan stain). The immuno-protective activity of the vaccines was determined by tumor growth inhibition and cytotoxic T lymphocytes (CTL) induction in vaccine immunized syngeneic mice. RESULTS: Substitution of cysteine by glycine at both positions 24 and 92 of HPV58 E7 abolished its transforming activity. Growth of HPV E7+ tumor in mice immunized with the recombinant vaccinia virus expressing HPV58 E7 was inhibited, and the surviving time of the immunized mice was prolonged. CTL activity was induced as revealed by in vitro cytotoxicity assay using E7+ tumor cells as target cells. CONCLUSIONS: HPV58 E7, with its transforming potential abolished, may be used as vaccine for immunotherapy of patients with HPV 58 related cancers. PMID- 12905607 TI - [Differential expression of the epithelial membrane protein 1 of laryngeal carcinoma]. AB - OBJECTIVE: In order to explore pathogenic mechanism of laryngeal carcinoma, the involved genes were identified in larynx carcinogenesis by comparing the gene expression profile in matched primary normal epithelial cells and primary laryngeal carcinoma cells from the same patients. METHODS: A cDNA microarray analysis consisting of 11,431 human genes revealed significant changes in the expression of 35 genes, with 8 genes being up-regulated and 27 being down regulated. The epithelial membrane protein-1 (EMP-1) is one of the down-regulated genes. EMP-1 expression in various kinds of laryngeal carcinoma was determined by semi-quantitative reverse transcriptase polymerase chain reaction (qRT-PCR). RESULTS: The EMP-1 mRNA levels in all laryngeal carcinoma cells was significantly lower than that in the matched primary normal epithelial cells (P < 0.05) and were not correlated to the stage and differentiation of laryngeal carcinoma (P > 0.05). CONCLUSIONS: The EMP-1 expression was correlated to larynx carcinogenesis and may be helpful to elucidate the pathogenic mechanism in laryngeal carcinoma. PMID- 12905608 TI - [Inhibition of ER alpha-mannosidase expression causes reduction and shortening of microvilli on rat liver epithelial cell WB-F344]. AB - OBJECTIVE: To study the effect of N-glycosylation on the modification of microvilli on the surface of rat liver epithelial cell WB-F344 and the growth of the cells in culture. METHODS: Recombinant adeno-associated virus (rAAV) expression vector pAGX (+) containing an antisense or a sense fragment of 6A8 cDNA encoding a human alpha-mannosidase was constructed. The recombinant vectors or the mock were transfected into WB-F344 cells by means of lipofectAmine. The transfected cells were selected in G418 medium and cloned by means of limiting dilution. Integration of the transfected DNA into host DNA was detected by neo PCR. Rat liver ER alpha-mannosidase activity in cell supernatant was measured by using P-nitrophenyl-alpha-D-mannopyranoside as a substrate. Microvilli on cell surface were observed upon a scan electron microscope. The growth curves of the cells in culture were drawn. RESULTS: The cell clones transfected with antisense 6A8 showed reduction of ER alpha-mannosidase activity with various degrees. Clone AS1 and AS2 cell showed a pronounced reduction of the enzymatic activity. In the study on AS1 cells, Con A binding to the cells was found to be enhanced, cell growth in culture became slow from day 5. The microvilli on the cells were reduced and blunted. CONCLUSIONS: Transfection with antisense 6A8 resulted in reduction and blunting of microvilli on the surface of growing WB-F344 cells, which might be related to N-glycosylation modification. PMID- 12905609 TI - [Isolation of a HBV-PreS2 epitope from a random peptide library displayed on the bacterial flagellin]. AB - OBJECTIVE: To establish an improved procedure for isolation and identification of epitopes from a random peptide library displayed on the bacterial surface. METHODS: Epitopes were screened from FliTrx random peptide library by a monoclonal antibody 3B9 against HBV-PreS2 protein. The enrichment was monitored in each round. Higher affinity clones were obtained by increasing the washing strength and randomly selected for sequencing and Western blot analysis. RESULTS: Clones specifically binding to antibody were enriched in each round. Ten sequences were obtained from sixteen sequenced clones, seven of them contained the common motif RXRGXY with high homogeneity to 135-140 amio acids in HBV-PreS protein and have positive results in Western blot analysis. The other three sequences have no typical motif RXRGXY and showed different Western blot results. CONCLUSIONS: It's easy and quick to drive epitopes from a random peptide library displayed on the bacterial surface. PMID- 12905611 TI - [The epidemiology of meconium stained amniotic fluid on hospital basis]. AB - OBJECTIVE: To explore the epidemiology of meconium stained amniotic fluid pero delivery. METHODS: 6,206 one-fetal delivery cases were divided into meconium stained amniotic fluid and normal amniotic fluid groups. Statistics were analysed on two groups using Cohort study. RESULTS: Incidence of meconium stained amniotic fluid was 16.4%. There was no significant difference on maternal medical complication between two groups. The percentage of parity, gestational week > or = 42 weeks and big birth weight were higher in meconium stained amniotic fluid group than that in normal amniotic fluid group (P < 0.001). The average Apgar score in meconium stained amniotic fluid group was lower than that in normal amniotic fluid group (P = 0.001). The percentage of stillbirth, low birth weight and transferred newborn care unit in meconium stained amniotic fluid group were higher than that in normal amniotic fluid group (P < 0.001). CONCLUSIONS: The relative factors on meconium stained amniotic fluid were maternal parity, gestational weeks > or = 42 weeks and big birth weight. No correlation between meconium stained amniotic fluid and maternal medical complication. Morbidity of newborn baby was higher in meconium stained amniotic fluid. Meconium stained amniotic fluid is a important clinical factor on evaluating pregnancy outcome. PMID- 12905610 TI - [Treatment of premenopausal patients with advanced or recurrent breast cancer by chemical ablation]. AB - OBJECTIVE: To evaluate the clinical efficacy and safety of luteinizing hormone releasing hormone (LH-RH) analog in premenopausal patients with advanced or recurrent breast cancer. METHODS: LH-RH analog (enantone 3.75 mg/2 ml) were administered to 28 premenopausal patients with advanced recurrent breast cancer and its efficacy and side effect were observed. RESULTS: The response rate (complete response and partial response) was 42.9%, and after 8 weeks of treatment, the plasma estrogen in all patients decreased to the level of postmenopause. No major adverse events were observed. CONCLUSIONS: LH-RH analog is effective and safe for premenopausal breast cancer with low adverse reaction and its administration method is easy. PMID- 12905612 TI - [Bone mineral and body composition analysis of whole body in 292 normal subjects assessed by dual X-ray absorptiometry]. AB - OBJECTIVE: To understand whole body bone mineral and body composition changes in normal subjects, and study the relationship between body composition and bone mineral. METHODS: 292 normal subjects aged 10-79 years old, including 140 males and 152 females, were selected to be measured bone mineral content (BMC), bone mineral density (BMD) lean and fat of whole body by dual X-ray absorptiometry (DXA). Individuals were divided into age-groups by every ten years and were analyzed by statistical methods. RESULTS: In males, peak values of BMC, BMD, lean and fat were in the 30-39, 20-39, 30-39, 70-79 age-groups. In females, they were in the 30-39, 30-39, 30-49, 50-69 age-groups respectively. Peak values of BMC, BMD and lean were higher in males than that in females, but peak value of BMD was not significantly higher in males than that in females. Peak value of fat was higher in females than that in males. Loss of BMC and BMD for females were more pronounced than that for males. Loss of lean for males was more pronounced than that for females. There are significant positive correlation between lean, weight and bone mineral in males and females. Fat has significant effect on BMC in females only. CONCLUSIONS: The normal bone mineral and body composition data of whole body for males and females, and the characteristic of changes with aging are provided for analyzing the relationship between bone mineral and body composition with ease. PMID- 12905613 TI - [The effect of estrogen benzoate on normal human breast tissue implanted into athymic nude mice]. AB - OBJECTIVE: To investigate the proliferating effect of estradiol benzoate on normal human breast tissue. METHODS: Three specimens of normal human breast tissue were implanted into nine 9-10-week-old intact female athymic nude mice which were randomly divided into group A, B and C. Each specimen was divided into three parts, and each part was implanted into mice of each group of three (A, B and C) respectively. Two weeks later, part of xenografts was taken out from group A as control group, and then group A, B and C were injected muscularly with 3, 6, 9 micrograms estradiol benzoate once daily, respectively. After therapy for 28 days, the grafts were taken out and expression of proliferating cell nuclear antigen (PCNA), estrogen receptor (ER) and proliferating receptor (PR) in grafts were examined using immunohistochemical method. RESULTS: Compared with control group, PCNA was increased in group B and C (P < 0.05), but in group A had no significant change. ER was gradually down regulated by 3, 6 and 9 micrograms estradiol benzoate (P < 0.05); however, PR was increasingly up regulated (P < 0.05). CONCLUSIONS: Certain dosage of estradiol benzoate has proliferating effect on normal human breast tissue. PMID- 12905614 TI - [Treatment of refractory irritable bowel syndrome with subclinical dosage of antidepressants]. AB - OBJECTIVE: To investigate the effect of antidepressant on irritable bowel syndrome (IBS). METHODS: A self-control and follow-up study on subclinical dosage of antidepressants therapy (fluoxetine 10 mg/d, paroxetine 10 mg/d or doxepin 45 mg/d) for 9-12 wks in 46 patients with refractory IBS symptoms according to Rome II criteria was performed, the clinical outcomes were evaluated by scales changes of symptom-related-anxiety, severity index of symptom, and quality of life specific of IBS, as well as general psychiatric health by SCL-90 during treatment and follow-up periods. RESULTS: All 46 cases completed therapy and first follow up unit (12 wks after treatment) (FFU), at the end of FFU, clinical symptoms in all patients were improved (P < 0.01). Comparison of the scores of symptom related-anxiety, index of symptom, and quality of life specific of IBS at the end of FFU with that at basal level, indexes of the severity (3.4 +/- 1.5 vs 1.8 +/- 0.84) and frequency (3.8 +/- 1.60 vs 2.0 +/- 0.76) of symptoms were subsided significantly (P < 0.01, respectively); the scores of symptom-anxiety questionnaire including body anxiety (16.04 +/- 1.65 vs 10.83 +/- 1.64, P < 0.001), cognitive anxiety (18.78 +/- 2.12 vs 11.17 +/- 1.89, P < 0.001), fear (15.80 +/- 1.76 vs 10.78 +/- 1.85, P < 0.001) and avoiding (15.47 +/- 1.53 vs 10.16 +/- 1.59, P < 0.001) were also subsided significantly. In the meantime, IBS QoL improved significantly (P < 0.05), dysphoria, body image, interference with activity, health worry, social reaction and overall scores were improved significantly (P < 0.01, respectively). The status of general psychiatric health was also improved significantly (P < 0.01). CONCLUSIONS: Treatment of refractory IBS with subclinical dosage antidepressant is rational and effective, However a further study on its mechanisms is suggested. PMID- 12905615 TI - [Binding characteristics of new synthesized opioid receptor ligands to cloned mu opioid receptors stably expressed in CHO cell]. AB - OBJECTIVE: To determine the affinity of new opioid receptor ligands to cloned mu opioid receptors stably expressed in CHO cell. METHODS: The binding characteristics of the opioid ligand [3H] diprenorphine (3H-dip) were studied by cellular biological techniques and radioligands binding in cloned mu opioid receptors stably expressed in CHO cells in saturation binding experiments, and were followed by competition binding experiments with a variety of new synthesized opioid receptor ligands. RESULTS: The Kd and Bmax of [3H] diprenorphine bound to mu receptors were 1.06 nmol/L and 930 fmol/mg protein, respectively. Competition binding experiments revealed that ligand 3# and 12# displayed much higher affinity than DAMGO and Morphine for the cloned mu opioid receptor. However, the affinities of ligands 2#, 6#, 8# and 9# were lower than DAMGO and Morphine. CONCLUSION: The present results suggest that the new ligands 3# and 12# have higher affinity to mu opioid receptors. However, ligands 2#, 6#, 8# and 9# have lower affinity to mu opioid receptors. PMID- 12905616 TI - [Recent advances in islet transplantation and pancreatic stem cell research]. AB - This paper reviewed recent advances in pancreatic islet transplantation research, including islet isolation, purification, culture, cryopreservation and immunoisolation. Latest progresses in induction of pancreatic stem cell and embryonic stem cell to differentiate into insulin-producing islets were also introduced. On the basis of the present situation and future development of islet transplantation-based therapies for diabetes, the author thought that allogeous islet transplantation is a main choice for type I diabetes today and pancreatic stem cell transplantation for tomorrow. PMID- 12905617 TI - [The study of biological properties of gliomas]. AB - In order to explore the new methods of biological treatment of human gliomas, this project is to study the biological properties of gliomas from four different aspects, the results show that there is a IL-6 autocrine loop in human gliomas and the growth of gliomas will be inhibited when the autocrine loop is broken. There is a magnificent predominant expression of Th2 cytokines in human gliomas and human glioma cells, the switching of Th2 to Th1 can inhibit the proliferation of glioma cells. The dosage of 100 micrograms/ml of erythromycin is the best of therapeutic effect. Angiostatin can not only inhibit the vascular endothelial growth, but also have the inhibitory role on the growth of glioma cells in vivo. The above studies have provided some new ideas and will be very helpful for the treatment of glioma patients. PMID- 12905618 TI - [Diagnosis and treatment of Paget's disease of the breast: report of 33 cases]. AB - OBJECTIVE: To study the clinical features and histologic criteria of Paget's disease of breast, and evaluate the potential prognostic factors for better appropriate treatment strategies. METHODS: Thirty-three patients treated for Paget's disease of breast were reviewed retrospectively. RESULTS: The median age of diagnosis was 57 years. Twenty-nine patients were treated by modified radical mastectomy, 4 cases by mastectomy. Twenty-three (69.7%) patients had underlying carcinoma. Twenty-six (78.8%) patients did not have a palpable breast mass, 10 of them was simple Paget's disease, eight had ductal carcinoma in situ, eight had invasive ductal carcinoma. Seven patients had a palpable breast mass, and none of them was simple Paget's disease, except one of them had ductal carcinoma in situ, six had invasive ductal carcinoma. CONCLUSIONS: Most patients with Paget's disease had associated breast carcinoma. Paget's disease of the breast with a palpable breast mass is more likely to be accompanied by an invasive ductal carcinoma and had a poor prognosis. Patients with any nipple complaints deserve a serious investigation. PMID- 12905619 TI - [Diagnosis and epidemiology of large intracranial artery stenosis]. AB - Extracranial carotid artery occlusive disease is a major cause of ischemic stroke in Caucasians. However, intracranial artery occlusive disease, especially middle cerebral artery (MCA) stenosis is more significant in Asians. The underlying mechanisms of ischemic stroke with intracranial artery occlusive lesions is different from that of extracranial artery it is expected to recognize the pathogenesis and epidemiology of intracranial artery occlusive disease. By digital magnetic resonance angiography (MRA) and transcranial doppler (TCD) instead of traumatic subtract angiography methods for screening diagnosis of intracranial artery stenosis that were developed in recent two decades. In current paper, we summarized the results studied with MRA and TCD in Peking Union Medical College Hospital and Chinese University of Hong Kong with literatures reviews in this field. Two aspects are discussed (1) Methods for diagnosis of intracranial artery stenosis; (2) Epidemiology of intracranial artery stenosis. PMID- 12905620 TI - [Non-diabetic renal disease in type II diabetes mellitus]. AB - OBJECTIVE: Several studies have suggested that non-diabetic renal disease (NDRD) was common among non-insulin dependent diabetes mellitus (NIDDM) patients with renal involvement. METHODS: The prevalence of NDRD among Chinese NIDDM population in PUMC hospital center was retrospectively studied. Renal biopsy specimens were evaluated with light-, immunofluorescence- and electron-microscopy. The cohort consisted of 33 NIDDM patients who received renal biopsy. RESULTS: Patients with both isolated diabetic nephropathy (DN, n = 7) and NDRD (n = 22) had comparable duration of DM, creatinine clearance, serum creatinine, albumin and 24 hours proteinuria, as well as incidences of retinopathy, neuropathy and hypertension. Significantly more patients with NDRD had haematuria (P = 0.030) or non-nephrotic proteinuria (P = 0.016). IgA nephropathy accounted for 40.9% of the NDRD identified. CONCLUSIONS: In this study, haematuria and non-nephrotic proteinuria predicted the presence of NDRD among NIDDM patients complicated with renal disease. IgA nephropathy is the most frequent type of NDRD in Chinese. PMID- 12905621 TI - [Progress in human disease proteomics]. AB - In the post-genomic era, with the accomplishment of the sequence mapping of human genome, one of the most important tasks for life science is the explanation and identification of human genome, that is, about 1/3 genes of human genome and their functions need further revealment and verification on the level of protein. In the field of functional proteomics, the human disease proteomics shows great potential in the discovery of new molecular targets and biomarkers for medicine and biopharmacy. In this article, we have made a concise discussion on the current status, existing problems and future development in the research of human disease proteomics both in and out of China. PMID- 12905623 TI - [Single nucleotide polymorphisms in CAPN10 gene of Chinese population and its correlation with type 2 diabetes mellitus in Han people of northern China]. AB - OBJECTIVE: To investigate the distribution of the single nucleotide polymorphisms (SNPs) in CAPN10 gene in Chinese population and their relation with type 2 diabetes mellitus in Han people of Northern China. METHODS: CAPN10 gene was sequenced to detect SNPs in 27 samples of different nationalities in China. 5 SNPs were genotyped with single-base extension (SBE) method to perform case control study in 156 normal Han people of Northern China and 173 type 2 diabetes and the 3 positive loci reported in the article were performed haplotype analysis. One positive locus was also analyzed with transmission-disequilibrium test (TDT) and sib transmission-disequilibrium test (STDT) in 68 type 2 diabetes pedigrees (377 cases). RESULTS: A total of 40 SNPs were identified in length of 8,936 bp, with an average of 1 in every 223 bp; The SNPs in CAPN10 gene did not distribute evenly and the SNPs in Chinese was different from that reported in American Mexicans. There was no significantly statistical difference in the allele frequency of the 5 SNPs between case and control (P > 0.05), and the haplotype frequencies in the two groups were not much different (P > 0.05). There was no positive results in TDT and STDT analysis (P > 0.05). CONCLUSIONS: The SNP distribution of CAPN10 gene varies with different nationalities. The studied SNPs in CAPN10 gene may not be the major susceptibility ones of type 2 diabetes mellitus in Han people of Northern China. PMID- 12905622 TI - [The association of two single nucleotide polymorphisms in PRKCZ and UTS2 respectively with type 2 diabetes in Han people of northern China]. AB - OBJECTIVE: To probe the candidate susceptibility gene (s) of type 2 diabetes in the formal mapping region, 1p36.33-p36.23, in Han people of Northern China using single nucleotide polymorphisms (SNPs). METHODS: 23 SNPs located in 10 candidate genes in the mapping region were chosen from public SNP domain by bioinformatic methods and single base extension (SBE) method were used to genotype the loci in 192 sporadic type 2 diabetes patients and 172 normal individuals to perform case control study. RESULTS: Among the 23 SNPs, 8 were found to be common in Chinese population. There were statistically different in the allele frequency of 2 SNP, rs436045 in the protein kinase C/xi gene and rs228648 in Urotensin II gene between case and control groups. CONCLUSIONS: The two SNP may be associated with type 2 diabetes in Han people of China, which makes base for further study of the relation between the genes they located with type 2 diabetes. PMID- 12905624 TI - [Fine mapping of susceptibility genes loci within chromosome 1 in Chinese Han families with type 2 diabetes]. AB - OBJECTIVES: To confirm previous whole-genome scan results of mapping type 2 diabetes susceptibility genes in chromosome 1 in Northern Chinese Han population by conducting a new genome scan with both an enlarged number of type 2 diabetes families and a new set of microsatellite markers. METHODS: A genome scan method was applied. After multiplexed PCR, electrophoreses, genescan and genotyping analysis, size informations for all loci were obtained, and a further study was done using both parametric and non-parametric linkage analysis to calculate the P values and Z-values of these loci. RESULTS: A total of 34 microsatellite markers distributed within 5 regions along chromosome 1 were surveyed, and 12,000 genotypes were screened. Evidence of linkage with diabetes was identified for 8 of the 34 loci (all the P-values of the 8 loci distributed in 3 regions were lower than 0.05, and the highest Z-value was 2.17). Interestingly, all the 5 markers at the P terminal 1p36.3-1p36.23 region, spanning a long range of 16.9 cM, suggested to be linked with the disease. The results of the other two regions were not consistent with the previous ones. CONCLUSIONS: The study results have confirmed those gained in the previous genome-wide scan. The fact that all 5 loci at the P terminal region displayed linkage with diabetes suggests that more than 1 susceptibility gene may reside in this region. PMID- 12905625 TI - [Differential expression of the genes in leukemia cell apoptosis induced by TRAIL]. AB - OBJECTIVE: To identify the genes differentially expressed in leukemia cell apoptosis induced by recombinant soluble tumor necrosis factor-related apoptosis inducing ligand (rsTRAIL). METHODS: Suppression subtractive hybridization (SSH) and polymerase chain reaction (PCR) were used for the cloning and identification of the genes differentially expressed in the apoptotic Jurkat cells induced by TRAIL. Slot blot and Northern blot were used for the expression pattern analysis of the genes. Automatic DNA sequencing was used for DNA sequence analysis. RESULTS: Six cDNA fragments differentially expressed in the Jurkat leukemia cells treated with TRAIL were found, in which four were inhibited and two were activated during the Jurkat cell apoptosis treated with TRAIL. Among which the five genes of A14, X1, D1, A23 and C5 were found at the first time by DNA sequencing and GeneBank database searching. So that they were registered in GeneBank as AW731601, AW731602, AW731603, AW731604 and BE239235, respectively. It was found that the gene D1 was expressed higher in Jurkat leukemia cells and MCF 7 breast cancer cells than that in K562 leukemia, 825 gastric cancer and 7721 liver cancer cells. CONCLUSIONS: Five novel cDNA fragments were found, and among which D1 might be a tumor specific gene. PMID- 12905626 TI - [A novel gene involved in blood glucose regulation]. AB - OBJECTIVE: To clone a novel gene relative to blood glucose regulation. METHODS: Rat modes of autonomous regulation of blood glucose was made by intra jugular vein to right atrium injection of high concentration of glucose solution, and the control rats were injected with 0.9%NaCl both before skeletal muscles were separated for gene analysis. The differentially expressed fragments were identified by differential display technology (DDRT-PCR). After slot blot and Northern blot analysis, the artificial positive fragments were excluded and the true EST (expression sequence tag) differentially expressed was obtained. These positive EST were used as probes to screen cDNA library of rat skeletal muscle. RESULTS: A novel full-length cDNA, named as Fang-2 was obtained. GenBank Accession No. was AF399874. Fang-2 was found rat homologue of human troponin T by blast software (NCBI). It shared 78% identical nucleotides, which showed the family proteins were conservative. After high concentration of glucose stimulation of rats, the expression of Fang-2 was down-regulated. CONCLUSIONS: A novel gene relative to blood glucose regulation was cloned from rat skeletal muscle. The gene can regulate blood glucose level by effect certain mechanisms unknown yet with down-regulation expression. PMID- 12905627 TI - [The cloning and prokaryotic expression of human tumor necrosis factor like protein]. AB - OBJECTIVE: To identify and clone the gene encoding human TNFLP (tumor necrosis factor like protein) for some functional study on TNFLP. METHODS: The full-length cDNA of TNFLP was isolated from fetal brain cDNA library. Several kinds of software were used to analyze nucleotide sequence and amino acid sequence of TNFLP. TNFLP mRNA distribution was identified by Northern blot. TNFLP-C and TNFLP N were expressed in E. coli with GST expression system. RESULTS: The cDNA of human TNFLP was 2,112 bp, which encoded protein of 208 amino acid. Hydrophobility analysis found there were two hydrophobility regions of human TNFLP. TNFLP-C (112 207 amino acid) and mouse TNF-alpha were homologous. The identity of their amino acid sequence was 42%. Moreover, both of them had a motif-TYKRL. TNFLP was located in chromosome 16. Human TNFLP was widely expressed in various human tissues. Northern blot showed TNFLP was highly expressed in heart, brain and spleen, only one transcript can be seen. GST-TNFLP-C and GST-TNFLP-N fusion proteins were obtained. CONCLUSIONS: Tissue expression spectrum of TNFLP and prokaryotic expression of TNFLP have been done, which establish the base for the functional analysis of TNFLP. PMID- 12905628 TI - [Cloning, expression and purification of neural specific HuD cDNA]. AB - OBJECTIVE: To prokaryoticly express and purify HuD protein and its RNA recognition motifs. METHODS: HuD protein was prokaryoticly expressed and purified by molecular cloning technology. Its biologic activity was testified by Western Blot. RESULTS: Purified HuD protein and its RNA recognized motifs were observed. CONCLUSIONS: The result might aid for basic research and clinical application. PMID- 12905629 TI - [The identification and cloning of human M961 full-length cDNA and its splicing isoform]. AB - OBJECTIVE: To identify and clone the gene encoding human M96 gene and study its expression spectrum in several blood cell lines. METHODS: According to the sequence of human EST which was highly homologous to the mouse M96 gene, primers used for library screening were synthesized, then the human adult testis and fetal brain cDNA library were screened. The gene was analyzed by making use of BLAST and CLUSTAL W, and its expression spectrum was studied by multiple-cell lines Northern blot analysis. The expression change of M961 in cell differentiation was observed by use of K562 cell line induced by hemin. RESULTS: Two cDNA clones encoding human M96 gene were isolated, identified and named as M961, and M962. They were found to be isoforms of each other. Northern, blot showed that M961 gene was expressed highly in CEM, Hel, Dami and K562 cell lines. However, during K562 cell line differentiation, process the expression of M961 elevated only slightly. CONCLUSIONS: M961 gene was expressed highly in pluripotent cell lines with erythrocytic and megakaryocytic potentials. PMID- 12905630 TI - [Annotation of complete genomic sequence of 3p24-p25 478 kb of human DNA]. AB - OBJECTIVE: To annotate the human genome 3p24-p25 478 kb complete sequence. METHODS: The protein-coding genes in the genomic sequence were identified by using ab initio gene finding, homology-based similarity database searching and all or partial mRNA aligning with genomic sequence, and the content feature of the genomic sequence were analyzed by using EMBOSS package. RESULTS: Two known genes SLC6A1 and SLC6A11 were identified; as well as the GC content of this genomic sequence was 47% and 3 putative CpG islands were predicted in the genomic sequence, located in 130,685-131,516 bp, 307,090-307,870 bp and 415,585-416,308 bp, respectively. CONCLUSIONS: The methods, as mentioned above, might be used for annotating the biological information in the genomic sequence, such as gene structure, GC content, CpG island. PMID- 12905631 TI - [Heat shock activated Rac-MEKK-JNK pathway and hsp90 beta gene expression]. AB - OBJECTIVE: To study the effect of Rac-MEKK-JNK (Rac-mitogen activated protein kinase kinase kinase-C-jun N-terminal protein kinase) signal pathway on heat shock-induced hsp90 beta gene expression and the impact of Hsp90 on the regulation of the pathway. METHODS: DN-Rac, DN-MEKK or DN-JNK were cotransfected with hsp90 beta CAT reporter plasmid beta 3.1 into Jurkat or LETPa-2 cells individually, the CAT mRNA expression was then determined quantitatively by competitive RT-PCR based system. Western blot was carried out to detect the expression level and phosphorylation of c-Jun in Jurkat and LETPa-2 cells that were transfected with DN-Rac, DN-MEKK or DN-JNK. By in vitro kinase activity assay and Western blot, the effect of geldnamycin (GA) on heat induced JNK activity were evaluated. RESULTS: In Jurkat cell transfected with DN-Rac, DN-MEKK or DN-JNK, heat shock induced relative CAT mRNA expression level was decreased to (72.8 +/- 5)%, (60 +/- 13.2)% and (47.7 +/- 12.1)% of the control respectively; while in LETPa-2 cell hsp90 beta 3.1 reporter gene expression was accordingly suppressed to (16.17 +/- 5.1)%, (50.2 +/- 8.7)% and (47.5 +/- 10)% of control. C Jun expression and phosphorylation were inhibited by the transfection of either one of DN-Rac, DN-MEKK or DN-JNK. With GA treatment, heat shock induced JNK activity was repressed, while the expression level of JNK or c-Jun was not obviously changed. CONCLUSIONS: Rac-MEKK-JNK pathway promotes heat shock induced hsp90 beta gene expression and hsp90 may participate in the regulation of heat shock activated Rac-MEKK-JNK signal pathway in both Jurkat and LETPa-2 cells. PMID- 12905632 TI - [Cloning cDNA encoding for GAGA-like element binding proteins in human Jurkat cells]. AB - OBJECTIVE: To explore GAGA-like element binding protein in human cells. METHODS: Yeast one-hybrid system was used to screen the GAGA-like element binding proteins in HTLV-1 transformed Jurkat cell cDNA fusion library. Total RNA extracted from Jurkat cells was first labeled by reverse transcription, and was taken as cDNA probe to hybridize with the candidate positive clones. RESULTS: 9 positive clones were obtained, and 6 out of the 9 clones were positively hybridized with the cDNA probe. CONCLUSIONS: 6 candidate clones encoding for GAGA-like element binding proteins were obtained from Jurkat cells for further investigation. PMID- 12905633 TI - [Activation of auto-mesenchymal stem cells of skeletal muscle by bone morphogenetic protein for rescuing bone marrow failure]. AB - OBJECTIVE: To explore the effect of bone morphogenetic protein (BMP) to activate mesenchymal stem cells of skeletal muscle for rescuing bone marrow failure. METHODS: The study was performed on lethal rat acute aplastic anemia model induced by combined 5-fluorouracil (5-FU) and busulfan. The rh-BMP-2 was implanted into the thigh muscle of the rats at 3 days before aplastic anemia was induced. In the control group the rats were implanted with agar into the thigh muscle. The blood picture, pathologic changes and the mortality in two groups were observed. At the same time, rh-BMP-2 were implanted into the thigh muscle of normal Kun-min mice for dynamic control observation of the implantation local morphological changes, colony forming units-spleen (CFU-S) and stem cell growth factor (SCF) expression of the stroma cells of ectopic ossicles induced by BMP. RESULTS: At 7 days after BMP implantation in the mice the mesenchymal cells around BMP in muscle proliferated, and appeared in bone marrow to form an ectopic ossicles. The SCF expression of stroma cells in ectopic ossicles were higher than that of self-bone marrow. 56.3% of BMP-treated aplastic rats were survived over 3 months and its hematopoiesis was completely reconstituted and the histo morphological picture of the spleen and bone marrow were recovered to normal. But in the control group only one of 23 rats was survived, the remainder died of hematopoietic failure. CONCLUSIONS: BMP-implantation into the skeletal muscle could rescue the bone marrow hematopoietic failure. The mechanism might be related to the BMP activated auto-mesenchymal cells of skeletal muscles to direct hematopoietic cell differentiation. In our hands it might create a new pathway for utilization of auto-muscle derived mesenchymal cells to reconstitute hematopoiesis. PMID- 12905634 TI - [Reconstitution of telomerase activity in human embryonic tendon cells transfected by ptsA58H plasmid]. AB - OBJECTIVE: To investigate the feasibility of life span extension of transformed human embryonic tendon cells (THETC) by reconstitution of the telomerase activity. METHODS: THETC were transfected by pGRN145 plasmid containing the human telomerase reverse transcriptase (hTERT) cNDA in vitro by molecular cloning technique. The biological characteristics of transfected cells were detected and compared by morphological observation, plate cloning efficiency, soft agar culture, growth curve of cells cultured in different conditions, immunohistochemistry, telomerase activity assay by telomeric repeat amplification protocol (TRAP). RESULTS: The THETC transfected by pGRN145 plasmid (telT) could express the telomerase activity with extension of life span. The telT maintained the original characteristics of temperature-dependant and serum-dependant, as well as secretion of type I collagen normally and without tendency of malignant transformation. CONCLUSIONS: The life span of THETC can be prolonged by reconstitution of telomerase activity, which provides the novel experimental methods to establish the standard cells line. PMID- 12905635 TI - [Effects of hydrogen peroxide on intracellular free Ca2+ content in rat liver oval cells]. AB - OBJECTIVE: To study the effects and mechanism of hydrogen peroxide (H2O2) of low concentration on dynamic changes of intracellular free calcium contents ([Ca2+]i) in cultural rat liver oval cells (WB-F344 cells). METHODS: Using Fluo-3/Am as fluorescent indicator of [Ca2+]i and it was measured by laser scanning confocal microscope system. RESULTS: The results showed that: (1) A rapid transient spiking of [Ca2+]i occurred after the stimulation of H2O2 of low concentration (800 nmol/L). (2) The [Ca2+]i increase was abolished by pretreated with catalase (CAT) or by incubated in D-Hank's solution containing EGTA, the chelate of extracellular Ca2+. (3) The [Ca2+]i increase was not inhibited by pretreated nifedipine, Ca2+ channel blocker, but was abolished by pretreated with anthracere 9-cardoxylic acid (A9C), the Cl-channel blocker and which also blocked calcium activated non-selective cation channel (CAN). CONCLUSIONS: These results suggest that the increase of [Ca2+]i induced by H2O2 of low concentration may be due to the influx of extracellular Ca2+ through CAN. PMID- 12905636 TI - [The role of p53 binding site on the trans binding of p53 to Hsp90 beta gene]. AB - OBJECTIVE: To investigate the effect of p53 binding site (+31/+60) of hsp90 beta gene on its transcriptional regulation. METHODS: The binding site was first inserted into pBS-SK. After the plasmid annealing and elongation with mutagenic and selective primers, nuclease digestion and bacteria transformation was performed twice to select the positive mutated plasmid. Electrophoretic mobility shift assays (EMSA) was employed to detect the binding of hsp90 beta gene fragment containing mutated p53 binding site and Jurkat cell nuclear extract transfected by p53 expression vector. RESULTS: The sequence analysis profile confirmed a successful mutation of two bases on the core sequence of the second half binding site. EMSA results showed the specific DNA-protein complex band disappeared after the mutation. CONCLUSIONS: The core sequence of p53 binding site plays a key role in the trans binding of p53 to hsp90 beta gene. PMID- 12905637 TI - [Roles of diaphragmatic crural barrier and esophageal body clearance in patients with gastroesophageal reflux disease]. AB - OBJECTIVE: Aim of this study was to investigate functions of lower esophageal sphincter (LES) barrier and esophageal clearance in fasting and postprandial stages in gastroesophageal reflux disease (GERD). METHODS: Eight patients with confirmed GERD and 8 healthy subjects (HS) were observed in the study. The esophageal pH and manometry were recorded simultaneously for 1 h during fasting and 2 h after a meal (1,675 kJ) using pneumohydrolic capillary perfusion system. RESULTS: (1) The esophageal pH monitoring showed that median of percentage of pH < 4 at postprandial in HS and patients with GERD was 0.45% and 11.2%, respectively (P < 0.05). (2) Pressure of lower esophageal sphincter (LESP) significantly decreased after a test meal in GERD (P < 0.001) and in HS (P < 0.001). (3) The amplitude of post-LES relaxation related to swallow (post-LESRA) in GERD was much lower than in HS either during fasting or postprandial stage (P < 0.05). (4) The tension of crural diaphragm at resting (Dia-A0) in GERD was lower than in HS during fasting and postprandial stage(P < 0.05). (5) The tension of crural diaphragm at deep inspiration (dia-AM) in GERD and HS increased 3 or 4 times at pressure at gastroesophageal junction (GEJ). (6) The peristaltic amplitude of the distal esophagus in GERD were much lower than that in HS in both pre- and postprandial periods. CONCLUSIONS: (1) Impaired clearance of Post-LESRA and esophageal body, function of diaphragmatic crural play an important role in development of GERD. (2) The tension of crural diaphragm at deep inspiration can increase the pressure at GEJ. PMID- 12905638 TI - [Complications associated with the technique of pedicle screw fixation]. AB - OBJECTIVE: To analyze the causes of the complications associated with the pedicle screw fixation during and after operation, as well as the methods for prevention and treatment. METHODS: From January 1988 to December 1999, 475 patients who underwent pedicle screw fixation for various spinal disorders were reviewed retrospectively with statistical analysis. RESULTS: Four hundred and twenty-eight patients (90.1% of all) were followed up for (6.0 +/- 3.7) years. Complications of CD, DRFS, Dick, RF and steffee among 72 cases (80% of all) occurred during the operation of pedical screw fixation in 26 cases with the incidence of 5.5% and after the operation in 64 cases with the incidence of 15%. The incidence of operative/postoperative complications were 0-12.9% in CD, 6.8%-11.9% in DRFS, 7.3%-12.7% in Dick, 2.4%-19.5% in RF, and 10.1%-21.5% in Steffee respectively. The main complications included pseudoarthrosis, pedicle screw broken, pedicle screw malposition and so on. The complications were mainly due to unskilled technique of pedicle screw fixation, implant defect in design and so on. CONCLUSIONS: The complications associated with the technique of pedicle screw fixation must not be neglected. The main causes are laid on unskilled technique of pedicle screw fixation and implant defect in design. PMID- 12905639 TI - [Serum levels of macrophage colony stimulating factor in the patients with Alzheimer's disease]. AB - OBJECTIVE: To investigate the availability of serum level of macrophage clony stimulating factor (M-CSF) as a marker for early diagnosis of Alzheimer's disease (AD). METHODS: The serum levels of M-CSF in 70 patients with AD, 52 healthy controls, 22 patients with VAD (vascular dementia) were measured and the serum levels of IL-1 beta, IL-6, TNF-alpha in 32 patients with AD and 20 controls were measured as well. RESULTS: Serum levels of M-CSF were significantly elevated in patients with AD when compared with healthy controls (P < 0.01) and VAD controls (P < 0.05) respectively. At the early stage of mild dementia and middle dementia, serum levels of M-CSF were significantly elevated, but at the later stage of severe dementia, they returned to normal level. Serum levels of IL-1 beta were significantly elevated in AD patients compared with controls (P < 0.05), and serum levels of TNF-alpha and IL-6 were within the normal range in patients with AD. CONCLUSIONS: The results suggest that serum M-CSF level may provide a convenient and sensitive means for the early diagnosis of Alzheimer's disease. PMID- 12905640 TI - [Precision of rat BMD measurements by dual-energy X-ray absorptiometry in vitro]. AB - OBJECTIVE: To evaluate the precision of rat bone mineral density (BMD) measurements by Norland Excellplus dual-energy X-ray absorptiometry (DXA) and to investigate the BMD changes in ovariectomized (Ovx) rats in vitro. METHODS: (1) The coefficients of variation (CV) for BMD measurements at various skeletal regions were repeatedly determined by DXA in 10 Wistar rats in vitro. (2) BMD in lumbar vertebra (L5) and both sides of femurs was measured in total 90 rats. And (3) changes in BMD between Ovx and sham rats were compared. RESULTS: (1) The short-term CVs of BMD measurements in different regions by DXA were as follows, 1.58% for lumbar vertebra (L5), 0.90% for left femur, and 0.86% for right femur, respectively. The long-term CVs were 2.22% for lumbar vertebra (L5), 1.09% for left femur, and 1.20% for right femur. (2) The BMD values in 90 Wistar rats were (127.5 +/- 12.3) in lumbar vertebra (L5), (82.6 +/- 11.3) in corpus vertebra (L5'), (150.7 +/- 10.6) in left femur and (149.9 +/- 10.6) mg/cm2 in right femur, respectively. The correlation coefficient of BMD measurements between left and right femurs was 0.792 (P < 0.001). (3) In Ovx group, the BMD values of corpus vertebra (L5') and distal femurs were significantly decreased, that was 10.0% 17.5% lower in comparison with sham group (P < 0.001). CONCLUSIONS: Measurement of rat BMD in vitro by Norland Excellplus DXA is a useful method, and it can reflect the changes in rat bone masses with good precision. PMID- 12905641 TI - [The efficacy of alendronate in the prevention and treatment of postmenopausal osteoporosis]. AB - OBJECTIVE: To investigate the effectiveness of alendronate Chinese national product in the prevention and treatment of postmenopausal osteoporosis. METHODS: The 56 postmenopausal women with osteopenia or osteoporosis were randomly divided into two groups: treated with alendronate 10 mg/d (28 cases) orally and placebo (28 cases), for 6 months. All subjects received 600 mg/d of calcium carbonate and vitamin D 1,000 U/d. Bone mineral density (BMD) of the lumbar spine, femoral neck, trochanter and Ward's triangle were measured by dual energy X-ray absorptiometry as well as the markers of bone turnover were analysed at the beginning and the end of the study. RESULTS: The results showed that lumbar spine BMD increased by 5% in the alendronate group (P < 0.01), but decreased in BMD of the lumbar spine and femur in the placebo group (P < 0.05) after 6 months of treatment. In the alendronate group the marker of bone resorption and bone formation were significantly decreased after alendronate therapy. There were no change in placebo group. CONCLUSIONS: Alendronate (Chinese national product) is effective in reducing bone turnover and promoting bone mass of postmenopausal osteoporosis. PMID- 12905643 TI - [PPAR gamma--the master of thrifty genes]. AB - Peroxisome proliferation is a cellular response to many chemical compounds affects including natural and modified fatty acids, phthalate and adipate ester plasticizers, leukotriene antagonists, acetylsalicylic acid and certain pathophysiological conditions including dramatic change of cellular morphology and enzymatic activity. Peroxisome proliferation phenomenon is seen primarily in liver and kidney. Hormones and nutritional factor can regulate peroxisome proliferation response. Sustained peroxisome proliferation can lead to hepatocarcinogenesis. The three types of peroxisome proliferator activated receptor, termed PPAR alpha, PPAR beta, and PPAR gamma, expressed in specific tissue, are consisted of a specific a nuclear receptor superfamily. After more than 10 years world wide research, the function of PPAR is clarified, as PPAR gamma, the master of thrifty genes, controls the expression of genes relative to adipogenesis, diabetes mellitus and obesity. The receptor is involved in transcriptional control of numerous cellular processes including cell cycle control, inflammation, immunoregulation and carcinogenesis. PMID- 12905642 TI - [The primary study on a novel protein binding to the death domain of the death receptor 4]. AB - OBJECTIVE: To clone and identify novel proteins binding to the death domain of the death receptor 4 (DR4). METHODS: The yeast two-hybrid system was used for this study. Automatic sequencing was carried out for DNA sequencing. The sequence homology and the functional domains were analyzed by BLAST and the ScanProsite Tool softwares, respectively. Co-immunoprecipitate method was used to confirm human formyl peptide receptor-like 1 (FPRL1) binding specifically with DR4CD (the cytoplasmic domain of DR4) in HEK293T cells. RESULTS: Two positive clones, named as pADB1 and pADB2, were obtained. BLAST searching showed that the homology of the insert sequence of pADB1 with the mRNA of FPRL1 was 97%. The insert of pADB2 shared no homology with any known peptides in GeneBank. Co-immunoprecipitate analysis further confirmed that FPRL1 could bind to DR4CD in vivo specifically. CONCLUSIONS: FPRL1 may associate with DR4CD in vivo specifically. The functional studies of FPRL1 in signaling pathway mediated by TNF-related apoptosis inducing ligand (TRAIL) are in active progress in our laboratory. PMID- 12905645 TI - [Clinical evaluation of two kinds homogenous assays for determination of high density lipoprotein cholesterol]. AB - OBJECTIVE: To evaluate the clinical efficacy of two kinds homogenous assays for direct determination of high-density lipoprotein cholesterol (HDL-C) based on the principle of polyanion polymer/detergent (PPD method) and polyethylene glycol modified enzyme (PEGME) method. METHODS: The two homogenous methods were compared with the precipitation method (PTA-Mg2+ method), their precision, accuracy, specificity and interference were also analyzed. RESULTS: Both homogenous HDL-C assays were precise, having a within-run CV < 3%, day-to-day CV < 3% and total CV < 4%. The HDL-C values measured by the two homogenous methods correlated well with those by PTA-Mg2+ method (X): Y = 0.9316 X + 0.1063, r = 0.9762 for PPD method (Y); and Y = 0.9106 X + 0.1368, r = 0.9894 for PEGME method (Y). The linearity studies showed the two homogenous methods to be linear up to 4.14 mmol/L. The lowest detectable concentration of the two methods was apparently 0.08 mmol/L. Recoveries of the two methods were 94.1%-106.2%. Hemoglobin did not interfere with the HDL-C results in the two homogenous methods, whereas icteric samples with total bilirubin > 200 mg/L showed discrepancies. Lipemia up to triglyceride concentration of 17.0 mmol/L did not interfere with the two homogenous HDL-C assays. CONCLUSIONS: The two new homogenous HDL-C assays meet the requirements for accuracy, precision, ease of handling with massive sample, allow full automation, and are clinically useful. PMID- 12905644 TI - [Establishment and application of a RT-PCR system for quantifying mRNA of 4 hsp genes in human cells]. AB - OBJECTIVE: To establish a RT-PCR system for detecting mRNA expression level of 4 hsp genes in human cells. METHODS: RT-PCR system was established with gene cloning, gene recombination, in vitro transcription and RT-PCR techniques; The detection of expression level of 4 hsp genes in SW13 cell was carried out with this system. RESULTS: In SW13 cell, hsp70 and hsp90 alpha were typical heat shock induced genes, while hsp60 and hsp90 beta were efficiently expressed and further induced by heat-shock to various extent. CONCLUSIONS: In our hands novel RT-PCR system can be used to detect mRNA expression level of 4 human hsp genes. PMID- 12905646 TI - [Pulmonary thromboembolism in systemic lupus erythematosus: clinical analysis of 11 cases]. AB - OBJECTIVE: To investigate the diagnosis and therapy of systemic lupus erythematosus (SLE) patients with pulmonary thromboembolism (PTE). METHODS: 11 hospitalized cases were reviewed retrospectively in PUMC Hospital during January 1984-July 2001. RESULTS: All 11 cases were suffered from severe active lupus with PTE. The SLE-DAI (SLE-disease active index) was 21.9 +/- 4.9. 7 cases had first onset of progressive Raynaud phenomenon. Anti-RNP antibody was positive in 73% of the cases. Echocardiogram revealed medium-severe pulmonary hypertension. When PTE was found, while 6 cases had started with smaller to medium dose of prednisone treatment, which was 20-30 mg/d, and other 4 cases received no prednisone. Only 1 received large dose of prednisone and immunosuppressor. Large dosage of prednisone, immunosuppressor with or without anticoagulant were given to those 6 and 4 patients after final diagnosis, respectively. 6 of 7 cases showed relieved Raynaud phenomenon while 4 cases hemoptysis were disappeared, echocardiogram had confirmed that pulmonary artery pressure decreased in 4 cases. [(31.7 +/- 12.4) mmHg]. 5 cases survived, 3 were dead and 3 failed to be followed up. CONCLUSIONS: Patients of SLE with PTE are liable to be misdiagnosed or missed-diagnosed, The risk factors are active-SLE, progressive Raynaud phenomenon, and symptoms of thromboembolism, positive anti-RNP antibody and mild-medium pulmonary artery hypertension. Combined therapy of present available measures like large dose of prednisone, immunosuppressors and anticoagulant are highly recommended. PMID- 12905647 TI - [Prospect in the study of creating new drugs from Chinese herbal medicine]. AB - This paper concisely describes the prospect in developing new drugs from Chinese herbal medicine. The search for bioactive constituents or leading compounds from natural sources especially from Chinese herbal medicine will be proved to be one of the most effective ways by changing the traditional concept of study on natural drugs using modern ideas. It is well known that plants in China provide a fertile source of natural compounds dealt with biological and structural diversity, which is the best channel to discover bioactive and leading compounds. Many instances have showed that structural transformation and analogues synthesis of known natural compounds or drugs can provide an important way to find new leading compounds. Study of the bioactive principle of Chinese herbal medicine involves an integration of chemical study and bioassay with intensive screening. It should be emphasized that the investigation of bioactive compounds for creating new drugs is the major task of Chinese pharmaceutical industry. PMID- 12905648 TI - [Effect of antisense thrombin receptor and p21 double gene co-expression system on the proliferation and apoptosis in human aortic smooth muscle cells]. AB - OBJECTIVE: To focus on the study of the effect on proliferation and apoptosis of human aortic smooth muscle cells (ASMC) by adeno-associated virus (AAV) vector carrying antisense thrombin receptor (ATR) and p21 double gene co-expression system. METHODS: Cultured human AMSC was infected with recombinant AAV containing ATR, p21 single gene and AP double gene respectively. The integration and expression of genes were confirmed by semi-quantitative RT-PCR. The anti proliferation effect was determined by MTT assay. Cell cycle and apoptotic cell counts were measured through Flow Cytometry. The rate of apoptotic cells was examined with acridine orange/ethidium bromide(AO/EB) stain. RESULTS: RT-PCR indicated that the exogenous genes had been integrated into ASMC. The rates of cell survival were decreased by 16.67%, 21.60%, and 29.4% and the cell counts of G0/G1 phase were (61.8 +/- 2.9)%, (82.5 +/- 4.0)%, (80.4 +/- 6.1)% in ATR, p21 and AP group respectively after rAAV infected 4 days. The level and area of apoptotic peak were greater in AP double gene than ATR and p21 single gene. Cell stain indicated that apoptotic cells were (7.2 +/- 3.3)%, (10.7 +/- 5.6)%, and (18.3 +/- 2.7)% in each transgene group compared with (1.5 +/- 0.8)% in control group. CONCLUSION: AP double gene co-expression system has powerful effect for inhibiting proliferation and inducing apoptosis ASMC than ATR and p21 single gene and that is a superior way for gene therapy to restenosis. PMID- 12905650 TI - [Effects of image quality of intravenous three-dimensional electron beam coronary angiography]. AB - OBJECTIVE: To identify reasons for poor image quality and nonassessability of coronary artery segments, and compare results between early and late diastolic triggering on coronary electron beam angiography (EBA). METHODS: One hundred patients referred for EBA were studied. Contrast-enhanced transaxial coronary images were acquired using electrocardiographic triggering and reconstructed three-dimensionally using volume rendering techniques. The image quality of coronary segments and image artifacts were analyzed statistically. RESULTS: Volume rendering was failed in 7 patients (7%) due to cardiac and breathing motions. Image quality was the best with the left main (LM), and worst with the left circumflex (LCX) coronary artery (P < 0.001). The image quality decreased systematically from proximal to distal within each coronary artery (P < 0.001). Forty percent R-R interval triggering on electrocardiography was better than 80% for image quality. The nonassessable segments occurred in 3% of LM, 2%, 8%, and 5% of proximal, 24%, 22%, and 12% of mid, 64%, 45%, and 20% of distal segments of the left anterior descending (LAD), LCX, and right coronary artery (RCA), respectively (P < 0.05). CONCLUSIONS: The major limitations of coronary EBA are in suboptimal spatial resolution and image artifacts. The image quality could be improved by using optimal electrocardiographic triggering. PMID- 12905649 TI - [Comparasion of the actions of human and porcine erythrocyte-derived depressing factor]. AB - OBJECTIVE: To compare the action mechanisms of human and porcine derived erythrocyte-derived depressing factor (h-EDDF and p-EDDF) as well as the effects on blood pressure. METHODS: The experiments were carried out in spontaneously hypertensive rats (SHR, n = 5) and two kidney-one click renal hypertensive rats (2K-1C, n = 7). The acute and chronic effects of h-EDDF and p-EDDF on blood pressure were observed, blood pressure test using tail plethysmography under unanaesthetic state. Both EDDF were administrated via jugular vein and/or oral respectively. The isolated thoracic aorta ring perfusion assay was used to examine the effect of EDDF on the asodilation. Primary cultured VSMCs were prepared from the thoracic aorta media of 2K-1C and normal Wistar rats. The effect of EDDF on proliferation of VSMCs were determined by MTT assay. The cell cycle of VSMCs was evaluated by flow cytometric. RESULTS: Both h-EDDF and p-EDDF could significantly decrease blood pressure of Wistar rats through intravenous administration and/or orally (P < 0.01 and P < 0.05 respectively). The contractile response of aorta in 2K-1C rats to PE was significantly enhanced compared with that of the control (P < 0.01) and both EDDF (10(-3) g/ml) remarkably induced a vasodilation with endothelium-dependent manner in SHR and 2K 1C rats (P < 0.05). h-EDDF and p-EDDF could significantly inhibit the proliferation of VSMCs from 2K-1C and control rats. After 24 hours of exposure to EDDFs the cell number of G0/G1 phase obviously increased and cell number in S phase was decreased (P < 0.01, respectively). CONCLUSIONS: It seems that the effects of h-EDDF and p-EDDF on blood pressure and vasodilation as well as inhibition of VSMCs proliferation and regulation of cell cycle have no significant difference. PMID- 12905651 TI - [Inter-population and inter-individual effect of education attainment on men's behavior of smoking: China multi-center study of cardiovascular epidemiology]. AB - OBJECTIVE: To study the inter-population and inter-individual effect of education attainment on men's behavior of smoking in China. METHOD: A cross-sectional study of 7,415 men, aged 35-59 years, from 15 population samples was carried out in 1998. Information on education attainment and smoking was collected by standardized questionnaires. The population samples were divided into 2 groups according to the proportion of men with college or above education attainment: group I had an average 25.8% of men with college or above education attainment, and group II had an average 2.0% of men with college or above education attainment. RESULTS: (1) In group I, the percent of ever smokers, current smokers, deeper smoker, and ever smokers with cessation due to disease and the average number of cigarettes per day for the smokers were significantly lower than in group II (66.2%, 56.4%, 34.4%, 47.8%, 17.2 cigarettes/day vs 75.8%, 64.4%, 40.3%, 58.4%, 20.4 cigarettes/day, respectively). (2) The percent of current smokers and the number of cigarettes decreased with increased education attainment in both groups. Taking the men with college or above education attainment in group I as reference, the odd ratio of smoking for those with high school education in group I was 1.8 (95% CI: 1.5-2.1), for those with middle school or lower education in group I was 2.0(95% CI: 1.7-2.5), for those with college or above education in group II was 1.4(95% CI: 0.9-2.1), for those with high school education in group II was 1.7(95% CI: 1.4-2.1), and for those with middle school or lower education in group II was 2.4 (95% CI: 2.1-2.8). CONCLUSION: Smoking behavior was affected by both individual education attainment and population education level. Smoking cessation measures should be focused on the development of better anti-smoking environment, such as education development, health promotion, etc. PMID- 12905652 TI - [An observation of taurine transport alterations in calcification of myocardial cells in vitro]. AB - OBJECTIVE: To observe the alterations of taurine transport, taurine transporter (TAUT) and cysteine sulfinate decarboxylase (CSD) mRNA in the calcification of myocardial cells in vitro. METHODS: 3H-taurine measured the amount of taurine uptake. TAUT and CSD mRNA consents were measured using competitive quantitative RT-PCR in cultured and calcified myocardial cells. RESULTS: In calcification of myocardial cells, taurine concentration was decreased by 27% (P < 0.05), taurine uptake was markedly reduced, Vmax reduced by 39% (P < 0.01), there were no statistical significance of Km values between the two groups. TAUT mRNA decreased by 45% (P < 0.01), but CSD mRNA increased by 25% (P < 0.05). CONCLUSIONS: The data suggest that there were impediment of taurine transport in calcification of myocardial cells, as TAUT mRNA level was decreased, but CSD mRNA concentration was improved. PMID- 12905653 TI - [Comparative study of pulmonary function after conventional ultrafiltration or modified ultrafiltration during cardiac surgery of infants]. AB - OBJECTIVE: To compare the effects of conventional ultrafiltration and modified ultrafiltration in protecting patients' pulmonary function during cardiopulmonary bypass. METHODS: Thirty infants patients (less than 7 kg) were divided into two groups: conventional ultrafiltration group (CUF, n = 15) and modified ultrafiltration group (MUF, n = 15). The volume of ultrafiltration, transfusion, hematocrit (HCT) before and after ultrafiltration, patients' respiration function (respiration index, A-aDO2, airway pressure), the time of mechanical ventilation and ICU in the two groups were respectively monitored. RESULTS: The transfusion in MUF group was significantly less than in CUF group (P < 0.01), and the volume of ultrafiltration in MUF group was significantly more than in CUF group (P < 0.01). The time of mechanical ventilation and ICU staying in MUF group were significantly shorter in MUF group than that in CUF group (P < 0.05). At 12 and 24 hours after operations, the A-aDO2 in MUF group was lower than that in CUF group (P < 0.05), and the respiratory index in MUF group was higher than that in CUF group (P < 0.05). CONCLUSION: The modified ultrafiltration can effectively improve pulmonary function after operations for low weight infants. PMID- 12905654 TI - [Study of autoantibodies against the G-protein-coupled beta 2- and alpha 1 adrenergic and AT1 receptors in patients with primary hypertension]. AB - OBJECTIVE: To determine whether autoantibodies against the cardiac G-protein coupled beta 2- and alpha 1-adrenergic and AT1 receptors are related to patients with primary hypertension. METHODS: Synthetic peptides corresponding to amino acid sequences of the second extracellular loops of the beta 2- and alpha 1 adrenergic and AT1 receptors were respectively used as antigens to screen sera from patients with hypertensive heart diseases (n = 50) as well as simple hypertension (n = 40) and healthy blood donors (n = 40) using ELISA test. RESULTS: The positive ratio of autoantibodies against beta 2 and alpha 1 and AT1 receptors in patients with hypertensive heart diseases were significantly higher than patients with simple hypertension and healthy donors. The geometric mean titers of autoantibodies against beta 2- and alpha 1-adrenergic and AT1 receptors had no difference between the patients with hypertensive heart diseases and the patients with simple hypertension, but the geometric mean titers of two groups were higher than healthy donors. In the patients with hypertensive heart diseases, 81.0% of the patients with autoantibodies against beta 2-adrenergic receptor had autoantibodies against alpha 1-adrenergic receptor and 76.2% had autoantibodies against AT1 receptors. The percent of the autoantibodies against three receptors in patients with hypertensive heart diseases were 52.4%. CONCLUSIONS: Autoantibodies against beta 2- and alpha 1-adrenergic and AT1 receptors play an important role in the pathophysiological changes of primary hypertension, and may participate myocardial and vessel remodeling. PMID- 12905655 TI - [Synthesis and preliminary studies of O-(2-[18F] fluoroethyl)-L-tyrosine as a positron emission tomography imaging agent]. AB - OBJECTIVE: To develop a 18F-labeled amino acid, O-(2-[18F]fluoroethyl) - L tyrosine(18F-FET), as a positron emission tomography (PET) tracer for imaging cerebral tumors. METHODS: 18F-FET was synthesized. Preclinical studies including sterility, endotoxin, and toxicity tests were performed. Two brain tumor cases were studied using 18F-FET and compared with 18F-FDG. RESULTS: Radiochemical purity of 18F-FET was over 95% which remained stable for 6 hours. The 18F-FET injection was sterile and its endotoxin content accorded with the standards of Chinese Pharmacopoeia. The uptake of 18F-FET in the normal brain tissues was significantly lower than that of the tumor, and the images of the brain tumor were clearer than those of 18F-FDG. CONCLUSION: 18F-FET can accumulate in the tumor tissues to give high quality images. It suggests that 18F-FET may be a safe and effective tracer for brain tumor imaging. PMID- 12905656 TI - [Construction of a collagen gel dermis with improved biological property]. AB - OBJECTIVE: To improve the biological property of artificial skin. METHODS: We have ameliorated Hansburgh and Middelkoop's method of manufacturing artificial dermis. The type I collagenase and Dispase were used to isolated neonate prepuce' dermis fibroblast. The gel dermis was constructed by compounding the fibroblast and collagen swelling solution. The property of the collagen gel dermis was measured. RESULTS: The neonate prepuce's dermis fibroblast had property of high proliferation, high activation of the dermis, and it could secrete abundant extracellular matrix (ECM). CONCLUSION: The collagen gel dermis is an useful dermis substitute. PMID- 12905657 TI - [Effect of alpha-difluoromethylornithine on growth characteristics and expression of ALT-04ag gene of human lung carcinoma cells]. AB - OBJECTIVE: To study the effect of polyamine biosynthesis inhibition on growth characteristics of human lung carcinoma cells and its correlation with the expression of human lung carcinoma associated antigen ALT-04ag gene. METHODS: The gene expression was detected by RT-PCR and immunocytochemical tests. The cell growth characteristics were studied by cell growth curves, morphological observation, FCM analysis and DNA electrophoresis. RESULTS: Human lung squamous carcinoma cells L78 treated with 5 mmol/L alpha-difluoromethylornithine (DFMO) for 5 days showed significant growth inhibition and apoptosis induction. The mRNA and protein expressions of ALT-04ag gene in the cells were downregulated, while these changes resulted from DFMO treatment were prevented by provision of DFMO along with exogenous putrescine. CONCLUSION: The effect of polyamine biosynthesis inhibition induced by DFMO restrains the growth characteristics and promotes apoptosis of human lung carcinoma L78 cells, which is associated with down regulation of ALT-04ag gene expression. PMID- 12905659 TI - [Architecture, influencing factors, and sensitivity to antifungal agents of Candida biofilm]. AB - OBJECTIVE: To establish a model of Candida biofilm and to explore its characteristics, ultrastructure, influences by saliva and serum, and sensitivity to antifungal agents. METHODS: Evaluations of the in vitro growth kinetics, influences by saliva and serum, and sensitivity to antifungal agents of Candida biofilm were performed with the abated tetrazolium salt XTT method on a 96-well microtire petri dish. The ultrastructure of Candida biofilm was observed under Confocal Laser Scanning Microscope (CLSM). RESULTS: The bioactivity of Candida biofilm increased with culturing time and serum could obviously increase the action of biofilm. The Candida biofilm was significantly resistant to routine antifungal agents. CONCLUSION: The Candida cells adhered in biofilms are significantly different in morphology from those in suspension and are resistant to routine antifungal agents such as Amphotericine B, Fluconazole and Itraconazole. PMID- 12905658 TI - [Expression and purification of gene transfer vehicle mediated by epidermal growth factor receptor]. AB - OBJECTIVE: To create a gene transfer vehicle for targeting gene therapy of cancer with epidermal growth factor receptor overexpressing. METHODS: Encoding sequences of the first domain of histone gene (H1) and EGF C-loop (EGFc) were obtained by PCR amplification. These two DNA fragments were ligated by EcoR I site, and cloned and sequenced. E. coli expression vector of the fusion gene was then constructed. The fusion protein H1EGFc was purified by specific band isolation from SDS-PAGE. RESULTS: The molecular weight of purified protein was consistent with the designed request. Its purity reached 94.02%. CONCLUSION: A fusion protein H1EGFc was expressed and purified. PMID- 12905660 TI - [Genotyping of HLA-DRB1 by PCR-SSP in Yunnan Lahu ethnic groups]. AB - OBJECTIVE: To investigate polymorphism of HLA-DRB1 in Chinese Lahu population in Yunnan. METHODS: Polymerase chain reaction-sequence specific primers (PCR-SSP) were used to determine HLA-DRB1 genotypes of 110 unrelated healthy Lahu minority people of Yunnan Province. RESULTS: Sixteen alleles of DRB1 were detected in this study. The results of test showed that the genotype distributions observed were corresponded with the Hardy-Weinberg equilibrium. CONCLUSION: This study has obtained a more comprehensive and accurate data set of the normal allele frequencies of HLA-DRB1 in Chinese Lahu population in Yunnan Province, which may be of significance in the studies on population genetics and disease association. PMID- 12905661 TI - [A mouse model of severe acute pancreatitis induced by caerulein plus lipopolysaccharide]. AB - OBJECTIVE: To set up a convenient nontraumatic mouse model of severe acute pancreatitis(SAP). METHODS: Mice received intraperitoneal injections with caerulein and lipopolysaccharide (LPS). Serum amylase and pancreatic moisture content were measured during experiment. The histo pathological changes of pancreas and relevant organs were observed under light microscope. RESULTS: Serum amylase and pancreatic moisture content increased and pancreatic interstitial edema, inflammatory cellular infiltration, parenchymal necrosis as well as parenchymal hemorrhages were happened in the caerulein plus LPS group, and the lesions of other organs including stomach, ileum, spleen, and lung were seen as well. In the careulein group, there was only pancreatic interstitial edema with no parenchmal necrosis or hemorrhage, and the rest organs were normal. CONCLUSIONS: The SAP mouse model induced by caerulein plus LPS has the same pathological characteristics of human SAP, which can be used for human SAP studies. PMID- 12905662 TI - [Identification and assessment of multiple human papillomavirus types in condyloma acuminata lesions from patients with genital warts in Beijing area]. AB - OBJECTIVE: To identify and assess multiple human papillomavirus types in condyloma acuminatum lesions from patients with genital warts in Beijing area, and compare different features between otherwise healthy and immunosuppressed patients. METHODS: PCR, RFLP and nucleotide sequencing analysis were used to determine HPV types from individual lesions. RESULTS: The predominant type from other healthy patients was HPV6, secondly HPV11. The mean age of patients infected by HPV6 was lower than that of HPV11 and HPV6 + 11. While lesions from immunosuppressed patients were often contained HPV11 or mixed with HPV6. Besides, HPV types 16 and 53 were detected from infected lesions than other HPV types. CONCLUSIONS: HPV6 was the major pathogen of condyloma acuminatum, but infected patients were at lower ages. While HPV11 was most often detected from immunosuppressed patients. As a low risk virus in normal genital tract, HPV53 also could be a pathogen in genital warts. PMID- 12905663 TI - [A forty-year study on hypertension]. AB - Since 1959 the investigations on prevalence of hypertension and studies on the prevention and treatment of this disease have been carried out. The vascular mechanism of hypertension and the depressor effect of Chinese traditional herbs were also studied in Chinese Academy of Medical Sciences. The results revealed that: (1) The prevalence of hypertension in Chinese adults increased from 7.73% in 1979 to 11.26% in 1991, both much higher than that in 1959 (5.11%). The rate of awareness, treatment and control was only 26.3%, 12.1%, and 2.8% respectively. The risk factors of hypertension included overweight and alcohol drinking. High sodium, low potassium, low calcium, and low animal protein diet were also very important risk for elevation of blood pressure. Hypertension was the most important causal risk factor of coronary heart disease and stroke. (2) Hypertension diagnosis and staging criteria were established in 1959. Secondary hypertension was found to constitute 1.1% among community hypertensive patients. The new concept of aortitis was formed and found to be the most common cause of renal vascular hypertension. Patient education together with low dose compounds of antihypertensive drugs was implicated widely. Randomized clinical trials Syst China, Post-stroke Antihypertensive Treatment Study, Chinese Acute Stroke Trial, and Chinese Cardiac Study 1 demonstrated benefits of treatment for hypertensive, stroke or acute myocardial infarction affordable by Chinese population at large. (3) A series of functional changes and abnormalities with evident hereditary characteristics were found in the processes of cellular Ca2+ transportation, utilization, metabolism and their modulation of the vascular smooth muscle in SHR, and SHRsp, which seem to be the principal cause of the increase in peripheral vascular resistance in hypertension. (4) Alkaloid of Rauwolfia verticilata and Ligustrazine had marked depressor effect. Flavones of Radix Pueraricae could reduce the cardiac and cerebral ischemic damage and symptoms in hypertensive patients. PMID- 12905664 TI - [Cardiovascular effects of estrogen in postmenopausal women]. AB - Cardiovascular disease continues to be the leading cause of morbidity and mortality among the postmenopausal women. Several randomized clinical trials to determine whether hormone replacement therapy (HRT) has a cardioprotective role and several others are currently underway. The results' however were controversial. During the past 3 years, a number of randomized clinical trials have investigated the role of HRT in primary and secondary prevention of clinical and anatomical manifestations of atherosclerosis and the clinical results suggest that HRT can not decrease cardiovascular events or slow down the development of atherosclerosis in women with established coronary heart disease (CHD). In contrast, HRT has been showed to possibly effective in women without CHD as primary prevention. PMID- 12905665 TI - [Determination of papaverine hydrochloride in skin and blood and the drug contents in pig skin]. AB - OBJECTIVE: To explore the application of ultraviolet spectrophotometer in determining papaverine hydrochloride content in blood and skin and to determine the intradermal content after local external application of papaverine hydrochloride. METHODS: Phosphate buffer solution (pH = 10, ionic strength = 0.4) was added into the blood or ground skin tissue containing papaverine hydrochloride, and papaverine hydrochloride was extracted with chloroform-hexane mixture (ratio:2:3). After oscillation and centrifugation, partial organic phase was retrieved and dried in 60 degrees C water bath. The dried product was dissolved in 1 mol/L HCl and then underwent filtration through 0.2 micron membrane. The ultraviolet apectrophotometry was applied to determine the content of papaverine hydrochloride in the filtrate. 2% papaverine hydrochloride cream was applied on pig's skin and skin specimen was retrieved at given time to determine the papaverine hydrochloride content in skin tissue with ultraviolet spectrophotometry. RESULTS: The absorbance spectrum of the extract was typical of that of papaverine hydrochloride, with no miscellaneous peak. The extractive rate of papaverine hydrochloride in blood of was more than 92% and that in skin tissue more than 87%. Eight hours after external application of 2% papaverine hydrochloride cream on pig skin, the content of papaverine hydrochloride in skin tissue was more than 90 micrograms/g, significantly higher than the effective concentration of papaverine hydrochloride. CONCLUSIONS: Ultraviolet spectrophotometry is a simple, economical and reliable method for the determination of papaverine hydrochloride content in blood and skin tissue, with good reproducibility and sensibility. External application of papaverine hydrocholine cream can provide efficient permeation and drug concentration in the skin. PMID- 12905666 TI - [Diagnosis and therapeutics of placental site trophoblastic tumor]. AB - OBJECTIVE: In order to get better understanding of placental site trophoblastic tumor (PSTT), we have investigated 4 PSTT cases that diagnosed in our hospital and with literature reviews. METHODS: We identified 4 patients of PSTT in our choriocarcinoma ward from Jan, 1985 to Dec, 2000 by retrospective analysis. RESULTS: PSTT accounted for 3.1@1000 of all trophoblastic diseases, the ratio of PSTT to choriocarcinoma was 1:138. The average age was 31. Two cases were secondary to hydatidiform mole with paremetrial or pulmonary metastasis respectively. All cases received hysterectomy and multiple chemotherapy. With a follow-up of 29 to 84 months, all patients are survived up to now. CONCLUSIONS: PSTT is a rare gestational trophoblastic tumor. Accurate pathological diagnosis is critical for proper treatment. The diagnosis and monitor are rather difficult because the beta-hCG are non-parallel to the growth and disappearance of the tumor. Operation plays an important role in its treatment, and multiple chemotherapy is helpful. PMID- 12905667 TI - [One-stage reconstruction of oral and maxillofacial soft tissue defect by using forearm free flap]. AB - OBJECTIVE: To study technique of one-stage reconstruction of oral and maxillofacial soft tissue defect by using forearm free flap. METHODS: The microsurgery procedure was used in the treatment of 4 oral cancer patients with oral and maxillofacial soft tissue defect after surgery. RESULTS: The forearm free flap was survived, for 4-18 months the oral function was well, and the appearance was satisfactory. CONCLUSION: This is a good approach to the reconstruction of oral and maxillofacial soft tissue defect by using forearm free flap technique. PMID- 12905668 TI - [Perioperative anesthetic management for the excision of phaeochromocytoma complicated with catecholamine cardiomyopathy]. AB - OBJECTIVE: To summarize experience of perioperative anesthetic management for patients undergone excision of pheochromocytoma and complicated with catecholamine cardiomyopathy. METHODS: Perioperative anesthetic management for surgical treatment of three cases of pheochromocytoma complicated with catecholamine cardiomyopathy was described and discussed according to literature reports. RESULTS: The catecholamine cardiomyopathy of the three cases presented with left ventricular hypertrophy, congestive cardiac failure and acute myocardial infarction. After removal of the pheochromocytoma under general anesthesia, a prolonged hypotension occurred in all of the three cases. In order to maintain stable hemodynamics, large dose of catecholamine was required after surgery. All of the three patients were survived and discharged. CONCLUSIONS: It is suggested that myocardial dysfunction may be another important factor for the prolonged hypotension after removal of the tumor. Meticulous preoperative assessment of heart function is of primary importance for the management of anaesthesia during surgical procedures. PMID- 12905669 TI - [Measurement of epidermal growth factor receptor concentration in the pre- and postoperative serum in patients with meningiomas]. AB - OBJECTIVE: To investigate effect of the serum concentration of epidermal growth factor receptor (EGFR) in the pre- and postoperative peripheral blood in patients with meningiomas. METHODS: Using ELISA, the EGFR concentration were measured in the pre- and postoperative serum in 53 patients with meningiomas. In this study, the patients were divided into preoperative group, postoperative group (including 42 total resection, 11 subtotal resection), and 28 healthy control. RESULTS: In 53 patients with meningiomas, concentration of preoperative serum EGFR was (352.93 +/- 66.18) fmol/ml, to show higher than control group (159.11 +/- 40.50) fmol/ml (P < 0.0001); Concentration of postoperative serum EGFR was (220.74 +/- 70.63) fmol/ml, to show lower than preoperative group (P < 0.001). Including in 42 patients with meningomas by total resection, serum EGFRs of the 38 patients were decreased (191.20 +/- 32.13) fmol/ml, the 4 patients with peritumoral edema were decreased (248.75 +/- 10.31) fmol/ml, to show lower than preoperative group (P < 0.001); the 11 patients with subtotal resection were decreased (322.14 +/- 89.53) fmol/ml, not to show different than preoperative group (P < 0.05). CONCLUSIONS: In 92.16% patients with meningiomas, the more poison of the tumor is resected, the less the serum EGFR concentration is detected postoperatively, while the total resection of mangionmas resulted in a lowest level of EGFR. These results support the concept that human meningiomas may be autocrine EGFRs. The measurement of serum EGFR can be useful to patients with meningiomas for follow up after surgery. PMID- 12905670 TI - [From clone selection to danger model]. AB - How is the balance between immune response and immune tolerance dynamically kept in the complicated immune system of human body? To answer this question, many scientists have proposed various models since 1950's. A brief introduction to these models is given in this mini-review, which might reflect the historical process in the development of immunology in the past half century. PMID- 12905671 TI - [Toll-like receptor and innate immunity in phylogenesis]. AB - In protection against microbes, an organism recognizes the pathogen associated molecular pattern (PAMP) on microbes by pattern recognition receptor (PRR). Toll like receptor is called innate immunity. A family of cell membrane receptor was found in recent years that can mediate innate immune responses through the activation of a series of immune-related genes. In phylogenesis, it is highly conservative. However, its functions are getting more diversified with the complication of the immune functions of organisms. PMID- 12905672 TI - [B cell activated co-receptor]. AB - B cell activated co-receptor plays important roles in linkage of innate and acquired humoral immune responses. CD21 molecule in the co-receptor complex is a receptor for C3dg and CD19 molecule enhances BCR signal transduction. CD21 also expresses on the surface of follicular dendritic cells, which mediates the long term maintenance of antigens and is indispensable for maintaining the memory of B cells. B cell activated co-receptor also has an effect on the negative selection of B cells reactive to autoantigens. PMID- 12905673 TI - [Cytotoxic T cell]. AB - Cytotoxic T cell (CTL) covers several subtypes, which are CD8+, CD4 and CD4-CD8-. CTL derives from T cell repertoire in lymphoid hematopoietic stem cells. It matures in thymus and is activated in peripheral lymphoid tissues. Effector CTL kills the target cells by 2 ways. One is apoptotic effect mediated by FasL-Fas pathway and the other one is cytolytic effect mediated by granzymes. CTL has aroused great attention due to its significance in anti-tumor and anti-virus. PMID- 12905674 TI - [Killing cell by granzyme]. AB - Granzyme is an effector molecule of activated cytotoxic T cells and natural killer cells. It mainly mediates cell apoptosis. Its function could be explained by its molecular characteristics to some extent. Its cytotoxic effect is related to some other factors contributing to apoptosis induction. It deserves studying if perforin mediates entrance of granzyme into cells. As potential substrates of granzyme caspases and their substrates have been paid much attention to. PMID- 12905675 TI - [Evidence-based clinical practice]. AB - Evidence-based medicine combines the patient's preferences with clinical experience and the best research evidence. Randomized clinical trials are considered the most valid research design for evaluating health-care interventions. However, empirical research shows that intervention effects may be significantly overestimated (in one study on average by about 50 per cent) in randomized clinical trials with inadequate methods for generation of allocation sequence, allocation concealment, and double blinding. Further, intervention effects may be significantly overestimated if trial results are not analysed by the intention-to-treat method. Unfortunately more than 90 per cent of the randomized clinical trials have inadequate randomization, lack double blinding, or do not use intention-to-treat analysis. These irregularities may explain the considerable variation of clinical practice-within regions, single countries, and single clinics. Accordingly, there is an urgent need to improve this situation. Guidelines for Good Clinical (Research) Practice, conduct of more trials as multicentre trials, The Consort Statement, and The Cochrane Collaboration may all help in the application of the best research evidence in clinical practice. By investments in education, applied research, and The Cochrane Collaboration, evidence-based medicine may form a stronger basis for clinical practice. PMID- 12905676 TI - [Evidence-based medical approach to clinical medicine and health policy-making in the 21st century]. AB - In the late part of the 20th century, due to the change of disease spectra, rapid increase of health expenditure and imbalanced distribution of health resources, the health management sectors and doctors increasingly demand the scientific decision-making. With the development of clinical evidences and research, evidence-based medicine (EBM) became emerged in the early 1990's. EBM, the medical science in compliance with the best available clinical evidence, integrates clinical epidemiology (CE), systematic reviews (SRs) and health technology assessment (HTA) as the major technology, to emphasize that the clinical practice should be based on the combination of the best available clinical evidence, medical experiences and patient desire to cope with the demand of medical decision-making. Some governments and international organizations like WHO, and Australia have now adopted the high-quality evidence of EBM for decision making. As the largest developing country, China is challenged by variety of health problems. The introduction, adaptation and implementation of EBM will improve the decision-making and health care services, which should enhance the competitive capacity of medical and pharmaceutical manufactures, as well as promote the popularization of EBM, and protect the doctors and patients rights. PMID- 12905677 TI - [Evidence-based medicine and clinical nutrition]. AB - Evidence based medicine (EBM), the medical science in compliance with the good clinical evidence, has boomed since its inception. Availing of three major technologies of clinical epidemiology (CE), systematic reviews (SRs) and health technology assessment (HTA), it emphasizes that clinical practice should be based on the high-quality clinical evidence. Clinical nutrition (CN) is an important medical field. The introduction of concept of evidence into clinical nutrition was started in the late 1980's. Along with the development of EBM, clinical practice of CN has increasingly applied with the principles of EBM. Now, 1,435 clinical trials related to CN have been identified in the Cochrane Library. Of Sichuan University, there still exists the gap in both research and practice of CN between China and the developed countries. The introduction and implementation of EBM into CN will be preferable to its improvement. The preparation of the Cooperating Group of Clinical Nutrition of the Chinese Cochrane Center by Clinical Nutrition Center of Peking Union Medical College Hospital and West China Hospital of Sichuan University will be a good attempt. PMID- 12905679 TI - [Meta-analysis of the relationship between smoking and stomach cancer]. AB - OBJECTIVE: To explore the risk of stomach cancer associated with cigarette smoking. METHODS: Meta-analysis was used to make formal summary statements about the relationship between smoking and stomach cancer. The results of the studies were analyzed by studied type(case-control study and cohort study) and gender(male and female). Homogeneity showed that random effect model should be selected to calculate the pooling relative risk (RR) and its corresponding 95% confidence interval (95%CI). RESULTS: The total pooling RR(95%CI) was 2.14(1.12 4.06), while the total pooling RRs (95%CI) of male and female smoking with stomach cancer were 1.70 (1.34-2.14) and 1.83 (0.10-33.09), respectively. CONCLUSIONS: Cigarette smoking is a risk factor for stomach cancer. Male smoking can increase risk of stomach cancer, however, the relationship between female smoking and stomach cancer should be studied further. PMID- 12905678 TI - [An appraisal of immunonutrition for clinical nutritional support with a systematic review of English and Chinese documents]. AB - OBJECTIVE: To evaluate the efficacy, safty and economical values of immunonutrition for clinical nutritional support. METHODS: The following electronic databases were searched: Chinese Biomedicine database (CBM), MEDLINE, EMBASE, Cochrane Library and SCI. Data were extracted by two reviewers. RevMan 4.1 was applied for statistical analysis. RESULTS: Seventeen randomised trials comparing the use of immunonutrition which comprises nucleotides, omega-3 fatty acids and arginine with standard enteral nutrition in surgical, trauma/burn and critical illness patients. Combined analysis indicated that the use of immunonutrition decreased infectious events (pooled OR was 0.51, 95%CI [0.38, 0.67], P = 0.00001), length of hospitalization, and the cost. In sensitivity analysis for mortality, we found an increased tendency in immunonutrition (pooled OR was 1.94, 95%CI [1.05, 3.57], P = 0.03). Further sub-group analysis indicated that the effects of immunonutrition were various in different patient types. CONCLUSIONS: Immunonutrition is associated with decreasing of infectious rates, length of hospitalization and cost in selective operation patients. But current evidence can't affirm the role of immunonutrition on critical illness patients. Further studies are urgently needed to clarify the reliability of immunonutrition in this group. PMID- 12905680 TI - [Meta-analysis of assisted music therapy for chronic schizophrenia]. AB - OBJECTIVE: To evaluate the effect of assisted music therapy for chronic schizophrenia. METHODS: 11 articles including 603 chronic schizophrenia patients were meta-analyzed using fixed effect model or random effect model. RESULTS: 6 randomized controlled trials were synthesized, showing that the difference was significant in statistics between experimental group (patients with music and drugs therapy) and control group (patients with drugs therapy only). Both the scores of SANS and BPRS for the control group were higher than those for the experimental group (SANS, d = 0.68; 95%CI: 0.46-0.90 and BPRS, d = 0.44; 95%CI: 0.06-0.82). In addition, the scores of both SANS and BPRS for the pre-test were higher than those for the post-test (SANS, d = 1.17; 95%CI: 0.02-2.32 and BPRS, d = 2.05; 95%CI: 0.28-3.82). CONCLUSION: The short-term effect of assisted music therapy is positive for chronic schizophrenia, but the long-term effect is still to be further studied. PMID- 12905681 TI - [Evidence-based clinical practice of radiotherapy of small-cell lung cancer]. AB - OBJECTIVE: To evaluate the effects of radiotherapy in the treatment of small-cell lung cancer and the optimal radiation doses, irradiation volume and fractionations. METHODS: Using evidence-based principles to search and evaluate clinical evidence on radiotherapy of small-cell lung cancer and giving grades of recommendation in clinical practice. RESULTS AND CONCLUSION: The combination of chemotherapy and thoracic irradiation were the treatment strategy in limited stage small-cell lung cancer (LD SCLC). There were no clear answers on optimal irradiation dose and volume. Early thoracic irradiations were better than later ones. Radiotherapy should be started at the first or second cycle of chemotherapy. Hyperfractionated irradiation may have therapeutic benefit compared with conventional irradiation. Prophylactic cranial irradiation could improve survival for patients with complete response after chemotherapy and radiotherapy. PMID- 12905683 TI - [Application of derivative Dolphe method in the constitution of practice guidelines on prevention and treatment of hypertension for primary care physicians]. AB - OBJECTIVE: To evaluate the necessity, scientificity, and feasibility of the drafted Practice Guidelines on Prevention and Treatment of Hypertension for Primary Care Physicians. METHODS: Derivative Dolphe Method was used to collect the responses from 50 experts in hypertension or related fields within the nation. An evaluation score of 1 to 5 was given to each item for selection, 1 for highly disagreed and 5 for highly agreed. The mean, standard deviation (SD) and coefficient of variance (CV) of the scores were calculated. RESULTS: (1) 90% experts responded to the evaluation sheet. (2) The mean score were above 4 for the five out of the six items related to necessity, the SD varied from 0.51 to 1.05, and the CV from 0.11 to 0.30. (3) Among the 28 items related to scientificity and rationality, the mean score was above 4 for 25 items and was between 3.5 and 4 for other three items. The CV was generally lower than 0.35. (4) Among the seven items related to feasibility, the mean score for five of them was lower than 4, and the CV varied from 0.21 to 0.33. CONCLUSION: The results indicate that the necessity and scientificity of the most content of the guidelines are confirmed by the invited experts. PMID- 12905682 TI - [A methodology study on assessment of cancer screening test]. AB - OBJECTIVE: To analyze the effect of gold standard, blind comparison and different cut-points choosing on screening techniques assessment, and to promote the application of evidence-based medicine theory in screening study. METHODS: A screening study for cervical cancer in rural China in 1999, where 1997 women had been tested for pathology as gold standard and simulating situations without gold standard, blind comparison and under different cut-points. Indices such as detectable rate, sensitivity and specificity were calculated for each technique. Receiver operating characteristics (ROC) curves were drawn and areas under ROC curves between screening techniques were tested. RESULTS: Without gold standard, diagnostic techniques could not be evaluated correctly, and without the blind comparison, the sensitivity and specificity of the tests would be subjectively increased. Furthermore, use of different cut-points led to different sensitivities and specificities of test. CONCLUSIONS: Gold standard, blind comparison and perfect cut-points can improve the quality of screening test and drawing ROC curves is an effective way to confirm cut-points and evaluate diagnostic techniques. It is necessary to enforce the application of evidence based medicine theory in scientific research. PMID- 12905684 TI - [Study on the function of HSD-3.8 gene encoding a testis-specific protein with yeast two-hybrid system]. AB - OBJECTIVE: To explore the protein factors that could interact with the testis specific protein encoded by HSD-3.8 gene (GenBank Accession Number AF311312) related with female fertilization. METHODS: Yeast two-hybrid system was used to screen the human ovary MATCHMAKER cDNA library with constructed "bait plasmid" containing the 0.7 kb fragment (HSD-0.7) of HSD-3.8. The interaction with the positive fragments using a series of truncated bait plasmids was investigated. RESULTS: One positive gene fragment was obtained, which coded for 144 amino acids of the C-terminus of human G protein beta subunit 1. Truncated bait plasmids couldn't interact with the fish protein fragment in yeast. CONCLUSIONS: The protein encoded by HSD-3.8 gene may function through G protein signal transduction pathway and the interaction depends on the integration of the bait protein. PMID- 12905685 TI - [Construction of recombinant human CD59 using ICAM-2 promoter for endothelial specific expression in xenotransplantation]. AB - OBJECTIVE: To construct a recombinant human CD59 gene containing intercellular adhesion molecule-2 promoter for high level endothelial-specific expression in xenotransplantation. METHODS: ICAM-2 promotor fragment and CD59-intron 1 fragment were produced by PCR from the human blood genome, and then clone these fragments into a pcDNA3-CD59 eukaryotic expression vector which was followed by digestion with the specific restricted endonuclease (for example: EcoRI, Hind III). The ICAM-2 promoter and CD59-intron 1 fragments were identified by PCR, and sequencing. The recombinant was then transfected into pig aorta endothelial cells with Lipofection, and the expression was measured by flow cytometer. RESULTS: Products of the sequences measured were in accord with the frames of the gene bank. The expression of the protein of this recombinant was positive. CONCLUSION: The CD59 recombinant gene is constructed successfully, providing a basis for transgenic research. PMID- 12905686 TI - [Vaccination of rhesus monkeys with recombinant antigen fragments and protection from hepatitis E virus infection]. AB - OBJECTIVE: To observe anti-HEV IgG response to vaccination of recombinant antigen fragments and evaluate its protection from Hepatitis E Virus infection in rhesus monkeys (Macaca mulatta). METHODS: Twelve monkeys were divided into three groups and immunized respectively with three different recombinant antigens: namely Ag1 (carboxyl terminal 431 amino acids of ORF2), Ag2 (128aa fragment at the carboxyl terminal of ORF2), and Ag3 (full length ORF3 ligated with two ORF2 fragments encoded by 6743-7126nt and 6287-6404nt). The monkeys were challenged intravenously with fecal suspension from experimentally infected rhesus monkeys, and the other three monkeys served as the placebo group for challenge with HEV. The dynamic changes of the levels of ALT and anti-HEV IgG were examined. Pathological changes of liver tissue were observed by light microscope. Excretion of virus was detected by RT-nPCR. RESULTS: Hepatic histopathology of two monkeys in the placebo group was consistent with acute viral hepatitis, and ALT was elevated 3-4 weeks after inoculated with virus, up to 10-20 times higher than normal level. The liver tissue of monkeys immunized with antigen kept normal, ALT in several monkeys elevated mildly, and anti-HEV IgG conversation occurred at 1-2 weeks after vaccination, with the titer reaching 1:12,800. The virus RNA could be detected by RT-nPCR from days 7 to 50 in monkeys of control group, and from days 7 to 21 in vaccinated monkeys after challenged with virus. CONCLUSIONS: The recombinant antigens could induce the production of anti-HEV IgG, which protected rhesus monkeys from acute Hepatitis symptoms related to HEV infection. PMID- 12905687 TI - [Establishment of drug-resistance cell line of human glioma mediated by MGMT]. AB - OBJECTIVE: To establish a drug-resistance cell line of human glioma mediated by MGMT. METHODS: Simulated the clinical usage of BCNU to establish a BCNU-resistant human glioma subline by cyclic exposing the U251 parent cells to a constant concentration of BCNU. The resistance index and the expression of MGMT mRNA of U251/BCNU were detected and compared the difference of in vitro proliferation between U251 and U251/BCNU. RESULTS: A subline--U251/BCNU was successfully established in about 4-month culture, which had a stable resistance to BCNU. U251/BCNU cells showed 17-fold higher resistance to BCNU than did U251 cells by MTT assay, while U251/BCNU cells expressed MGMT mRNA. The doubling time of U251 and U251/BCNU had no statistical difference. CONCLUSION: A drug-resistance cell line of human glioma mediated by MGMT is established, which could provide experimental basis for further studies on the resistance mechanism and reversal methods of glioma chemotherapy. PMID- 12905688 TI - [Expression of rat protamine gene in MEL cells]. AB - OBJECTIVE: To investigate the expression of rat protamine (RP) gene in MEL cells and the effect on cell growth. METHODS: Eukaryotic expression plasmid pCR-3.1-RP was constructed and transfected into MEL cells. The changes of cell growth rate, mitotic index and colony-forming rate in semi-solid medium were investigated. RESULTS: Transfected MEL cells showed lower growth rate, mitotic index and colony forming rate. Volumes of cells were reduced and reduction of RNA transcription was observed. CONCLUSION: These results suggest that expression of RP in MEL cell may inhibit the cell growth and proliferation. PMID- 12905689 TI - [Hemangioblastic characteristics of fetal bone marrow-derived Flk1+ CD34- cells]. AB - OBJECTIVE: To investigate whether fetal bone marrow stromal cells have hemangioblastic characteristics. METHODS: Human fetal bone marrow stromal cells (hfMSCs) were isolated and cultured. Immunophenotypes of hfMSCs were tested by FACS. hfMSCs seeded in the matrigel were induced with vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) in vitro. Vascularization and hematopoiesis were detected with immunohistochemistry and electron microscope. RESULTS: The typical properties of this CD34- stromal cell population were that 99% cells expressed Flk1 (vascular endothelial cell growth factor receptor 2) and tube structure was formed. In the process of induction, hfMSCs could give rise to CD34+ round cells. CONCLUSIONS: We have demonstrated that fetal bone marrow stroma-derived Flk1+ CD34- cells could differentiate into vascular endothelial cells and hematopoietic cells, indicating that fetal bone marrow stroma-derived Flk1+ CD34- cells have hemangioblastic characteristics. PMID- 12905690 TI - [Serial analysis of gene expression in the pituitary adenomas and para-tumor normal pituitary tissues]. AB - OBJECTIVE: To observe the characteristics and difference of gene expression in the pituitary adenomas and para-tumor normal pituitary tissues. METHODS: Using serial analysis of gene expression (SAGE), two SAGE libraries were generated. Forty clones from each SAGE library were sequenced, and the results were analyzed by SAGE2000 software and compared with the SAGE map at NCBI. RESULTS: A total of 655 gene tags, representing 43 genes, were extracted from the 40 sequence files of the para-tumor normal pituitary tissues and 737 gene tags, representing 53 genes, were extracted from the 40 sequence files of the pituitary adenomas. Of these tags, 13 were not reported before. The genes related to pituitary hormone secretion and energy metabolism were highly expressed in the two kinds of tissues. Some growth factors and cytokines were also expressed, including those involved in the immunological system. But there were also much difference of gene expression in the two tissues. Thirty-one and five tags were only detected in para-tumor normal pituitary tissues and pituitary adenomas, respectively. CONCLUSIONS: Genes involved in hormones secretion and energy metabolism were highly expressed in the pituitary adenomas and para-tumor normal pituitary tissues. Many growth factors and cytokines were also expressed in pituitary. There was also much difference of gene expression in the two kinds of tissues. SAGE can be used not only in understanding the quantity information of gene expression, but also in finding new genes. PMID- 12905691 TI - [Culture and identification of dendritic cells from the peripheral blood of patients with laryngeal squamous cell carcinoma in vitro]. AB - OBJECTIVE: To culture dendritic cells (DC) from peripheral blood of patients with laryngeal carcinoma for therapeutic aid. METHODS: Adherent peripheral blood mononuclear cells from peripheral blood were cultured with 15 ng/ml rhGM-CSF and 7 ng/ml rhIL-4 for one or two weeks. The purity of DC was detected by immunocytochemistry method. The mixed leukocyte reactions stimulated by DC loaded with laryngeal carcinoma antigen were tested by measuring 3H-TdR uptake. RESULTS: A considerable number of suspended cells with spicular or dendritic appearance were observed after 1 week of culture, and their mitochondria were rich in cytoplasm. The positivity of DC was about 30%-60%. DC loaded with laryngeal antigen could induce proliferation of syngeneic T lymphocytes. CONCLUSION: A large number of DC with high purity can be cultured from peripheral blood of patients with laryngeal carcinoma in vitro. It may be used in further experimental studies for clinical applications. PMID- 12905692 TI - [Genetic polymorphism of 9 STR loci in Han nation population in Shandong Province]. AB - OBJECTIVE: To study genetic polymorphism of 9 STR loci in Han nation population in Shangdong Province. METHODS: We investigated 100 unrelated individuals of Han nation population in Shandong Province and studied their genetic distribution of 9 STR loci and Amelogenin locus. Primers of 10 loci were labeled with the NHS ester fluorescent dye 5-FAM (blue), Joe (green) or NED (yellow). The data of Han nation population were generated by multiple amplification and subjected to GeneScan, genotype and genetic distribution analysis. RESULTS: 83 alleles and 220 genotypes were observed, with the corresponding frequency of 0.0050-0.4050 and 0.0100-0.2100. The average of heterozygosity was 0.7778, the accumulated discrimination power was 0.9999. The accumulated probability of exclusion paternity was 0.9999. The polymorphism information content was 0.5823-0.8396. CONCLUSIONS: Chi-Square test indicated that the distribution of genotypes agreed with Hardy-Weinberg equilibrium. PMID- 12905694 TI - [Relationship of changes in sleep architecture and cognitive function in patients with obstructive sleep apnea syndrome]. AB - OBJECTIVES: To explore the relationship between sleep architecture changes and cognitive impairment in patients with obstructive sleep apnea syndrome (OSAS). METHODS: Thirty-six patients with OSAS and 18 controls were administered polysomnography and neuropsychological tests of visual regeneration, digital symbol, comprehensive memory and digital span. RESULTS: Performances on the visual regeneration test and digital symbol test in patients with OSAS were impaired significantly compared with the control group (P < 0.01). Patients' performance on the visual regeneration test was significantly associated with slow wave sleep duration (r = 0.423, P < 0.05) and that on the digital symbol test was significantly associated with REM sleep duration (r = 0.378, P < 0.05). CONCLUSIONS: Slow wave sleep and REM sleep derivation may play a role in the cognitive impairment in patients with OSAS. PMID- 12905693 TI - [Association of polymorphism of human beta 2-adrenergic receptor gene and bronchial asthma]. AB - OBJECTIVE: To investigate whether beta 2-adrenergic receptor gene (beta 2AR) polymorphism at position 16, 27, 164 is in association with asthma susceptibility or asthmatic phenotype (including nocturnal asthma, serum IgE level, bronchial responsiveness, the status of asthmatics). METHODS: By using PCR-RFLP and allelic specific PCR (ASP), the polymorphism of beta 2AR gene at position 16, 27, 164 in 125 Han origin asthmatics and 96 normal healthy controls with the same ethnic nearby Beijing region were genotyped. All patients had their serum total IgE (TIgE) measured by RAST, pulmonary ventilatory function assessed by FEV1% and FEV1/FVC, bronchial responsiveness challenged by methacholine (if FEV1% > 70%), and brocho-reversibity by inhaling beta 2-agonist. RESULTS: There was higher prevalence of Gly16 homozygous of beta 2AR in asthmatics than that in normal healthy controls (22.4% vs 8.3%, P < 0.05), with odd ratio (OR) 2.918 (95% CI: 1.256-6.781); Also there was higher frequency of Gly16 homozygous of beta 2AR in nocturnal asthmatics than that in nonnocturnal asthmatics (35.3% vs 13.5%, P < 0.01), but Gly16 homozygous of beta 2AR was low an independent risk factor for the pathogenesis of asthma. The dose of methacholine was low in asthmatics carrying Gln27 homozygous beta 2AR than Glu27 homozygous beta 2AR and Gln/Glu27 heterozygous beta 2AR in brocho-challenge test [(0.205 +/- 0.275) vs (2.11 +/- 3.00) vs (1.575 +/- 0.828) mumol, P < 0.05]. CONCLUSIONS: Gly16 homozygous beta 2AR was associated with asthma susceptibility in Chinese patients with Han ethnic nearby Beijing region, and Gly16 homozygous beta 2AR was associated significantly with nocturnal asthma. Glu27 homozygous beta 2AR was related to hyper-bronchial reactivity of asthmatics. PMID- 12905695 TI - [Effects of sex hormones on serum levels of nitric oxide and plasma angiotensin II in postmenopausal women]. AB - OBJECTIVE: To observe the effect of estrogen and progestin on the blood levels of nitric oxide and angiotensin II in aid of the application of hormone replacement therapy in postmenopausal women. METHODS: The serum nitric oxide and plasma angiotensin II levels in postmenopausal women were determined before and 3 months after oral intake of estradiol valerate 1 mg/day (n = 10) or estradiol valerate, 1 mg/d plus medroxyprogesterone acetate, 2 mg/d (n = 30). RESULTS: The serum nitric oxide levels of postmenopausal women were significantly increased by 3 months of oral estradiol valerate 1 mg/d (P < 0.05), whereas the plasma levels of angiotensin II tended to decrease. The positive correlation between the increases of nitric oxide and the changes of estradial 3 months after oral intake of estradiol valerate 1 mg/d was significant. Compared with the baseline, no significant changes were observed in both serum nitric oxide levels and plasma angiotensin II levels 3 months after oral intake of estradiol valerate, 1 mg/d plus medroxyprogesterone acetate, 2 mg/d (P < 0.05). CONCLUSIONS: The vascular functions can be improved through increasing the serum nitric oxide level after 3 month oral intake of estradiol valerate, 1 mg/d in postmenopausal women, and estradiol valerate plus medroxyprogesterone acetate intake may attenuate the beneficial effects. PMID- 12905696 TI - [Effect of premenopausal hysterectomy on ovarian function]. AB - OBJECTIVE: To investigate the influence of premenopausal hysterectomy on the function of the conserved ovaries. METHODS: A retrospective survey was conducted by sending questionnaire to 521 cases of hysterectomized women (with the conservation one or two ovaries) for benign gynecological diseases and 1,600 cases of normal controls (with contact uterus and bilateral ovaries). Age, type of operation, diagnosis and the time of the onset of the menopausal syndrome were asked in the questionnaire. RESULTS: The mean age of the onset of the hot flush (293 cases) in the hysterectomized group was (44.64 +/- 4.31) years, which was significantly lower than that of the normal group [(46.87 +/- 4.22) years, 197 cases] (P < 0.01). The mean age of the above 293 women undergoing hysterectomized was (42.84 +/- 4.37) years. The range of the operation was positively correlated with the time of the onset of the menopause syndrome. So there was only (1.99 +/- 2.40)-year interval between the hysterectomy and the onset of the hot flush. CONCLUSION: Hysterectomy with the conservation of bilateral/unilateral ovaries may have some certain influence on ovarian function. PMID- 12905697 TI - [Follow-up studies in polymyalgia rheumatica and temporal arteritis]. AB - OBJECTIVE: To analyze the clinical course and duration of therapy in 28 patients with polymyalgia rheumatica (PMR) and/or temporal arteritis (TA) for identifying factors that influence prolonged steroid use and relapses. METHODS: 28 cases of PMR and/or TA diagnosed from 1992 to 2001 were retrospectively studied in PUMC hospital. Patients were grouped according to the absence or presence of corticosteroid resistant and relapses. RESULTS: Of 28 patients, 22 had pure PMR, 3 had both PMR and TA and 3 had pure TA. 15 patients received corticosteroid therapy and 13 had both corticosteroid and immunosuppressor therapy. The median duration was (25.5 +/- 24.0) months. Increase of white blood cell level and higher baseline erythrocyte sedimentation rate (ESR) were significant risk factors associated with corticosteroid resistant (P < 0.01). Quicker reduction of corticosteroid dose was associated with relapse (P < 0.05). CONCLUSIONS: Patients with increase of white blood cell level and higher baseline erythrocyte sedimentation rate (ESR) are more likely to be resistant to corticosteroid therapy. Quicker reduction in corticosteroid is more likely to relapse. Immunosuppressor therapy should be added to patients who has corticosteroid resistant, and relapse or PMR associated with TA. PMID- 12905698 TI - [Intra-arterial thrombolysis in patients with acute ischemic stroke of the cervical internal carotid artery occlusive disease]. AB - OBJECTIVE: To observe the safety and efficacy of local intra-arterial thrombolysis (LIT) in patients with acute ischemic stroke of cervical internal carotid artery occlusive disease. METHODS: 10 patients were treated by LIT. 2 of 10 patients were further treated by either angioplasty or endarterectomy. Primary neuroradiological assessment was performed with CT in all patients. Angiographic recanalization level was classified according to thrombolysis in myocardial infarction (TIMI) grades. Clinical outcome was classified as good for Modified Rankin Scale (MRS) scores of 0 to 3 and as poor for MRS scores of 4 to 6. RESULTS: Complete/partial recanalization after infusion was accomplished in 8 patients, seven of whom had good clinical outcome. Cerebral hemorrhage occurred in 2 of 10 patients. CONCLUSION: The results of this study suggest that LIT may be helpful to improve the outcome in patients with acute cervical internal carotid artery occlusive disease if it is performed within 6 hours of stroke onset. PMID- 12905699 TI - [NK cell receptors and related ligands]. AB - NK cell has antigen-non-specific receptors on its surface. Based on the molecular structure, NK cell receptors can be divided into two types, including Ig superfamily and C-type lectin superfamily. Based on function, NK cell receptors can be divided into activation receptors (NKAR) and inhibition receptors (NKIR). NKAR includes CD16 mediating antibody dependent cytotoxicity, NKR-P1 mediating natural cytotoxicity, KAR conjugated with DAP12 and co-stimulatory receptor. They transduce signal through the ITAM motif in their cytoplasmic region. NKIR mainly include KIR and CD94/NKG2. Their ligands are MHC-I molecules. They are closely related to the recognition of auto cells by NK cells. Inhibition of NKAR activation and cytotoxic effect of NK cells by NKIR are carried out through the ITIM motif and require co-aggregation of NKAR and NKIR. PMID- 12905700 TI - [CD94/NKG2A--a kind of inhibitory receptor belonging to C-type lectin superfamily]. AB - CD94/NKG2A is a kind of inhibitory receptor belonging to C-type lectin superfamily. It specifically expresses on the surface of some lymphocytes such as NK cells and T cells, and mediates inhibitory signal. In this mini-review, the structure of CD94/NKG2A molecule is described and its biological significance is discussed. PMID- 12905701 TI - [Analysis and synthesis: retrospection and consideration of medical genetics]. AB - The history of medical genetics was reviewed in this article, through formal genetics, cytogenetics and biochemical genetics to molecular genetics. The application of analysis and integration in medical genetics is summarized, and the blueprint of medical genetics is also discussed. PMID- 12905702 TI - [Establishment of BAC mediated transgenic mice containing 97 kb beta-globin gene cluster]. AB - OBJECTIVE: To delete IL-11 receptor alpha chain gene from the Bacterial Artificial Chromosome (BAC) chimeric DNA by RecA protein mediated homologous recombination method and establish the transgenic mice model containing whole beta-globin gene cluster. METHODS: Two 500 bp homologous sequences (A and B) located at the upstream and downstream of IL-11 receptor alpha chain gene respectively were cloned into the Hind III and Xba I sites of pBV vector, then the 1 kb A + B fragment was recovered from the building vector and inserted into the Sal I site of the shuttle vector pSV-RecA. After transforming the shuttle vector into the competent DH10B E. Coli containing BAC DNA, the IL-11 receptor alpha chain gene was finally deleted from the BAC DNA through chloramphenicol positive selection and fusaic acid negative selection. The new BAC clone was characterized by Pulse Field Gel Electrophoresis (PFGE). Then, we microinjected the linearized and purified BAC DNA into the mouse fertilized eggs and prepared the transgenic mice. RESULTS: By RecA protein mediated homologous recombination method, we deleted the IL-11 receptor alpha chain gene from the BAC DNA containing the complete beta-globin Gene Cluster and established 3 respective transgenic mice lines. CONCLUSION: Human beta-globin gene cluster in the transgenic mice mediated by new BAC expresses in a correct mode and level as compared with previous transgenic mice. PMID- 12905704 TI - [Analysis of human cells in transplanted goats using fluorescence in situ hybridization]. AB - OBJECTIVE: To analyze the existence and the dynamic cell frequencies of human cells in goats transplanted in utero with human hematopoietic stem cell (hHSC) by using fluorescence in situ hybridization (FISH) technique. METHODS: Interphase FISH (IFISH) with human-specific 17-chromosome satellite DNA and/or human specific Y-chromosome satellite DNA as probes was performed to analyze the presence and proportions of human cells in 13 transplanted goats. Samples were peripheral blood cells, bone marrow smears and liver touch imprint preparations. RESULTS: Of the 13 transplanted goats, eleven were identified to present human cells. Among them, two goats transplanted with human male HSC were found to have human male cells. The results demonstrated that these transplanted goats were human/goat HSC xenogeneic chimeras. Human cell frequencies decreased with the goat age (months), but the longest survival reached 21 months. During the detected life periods of goats, human cell frequencies in peripheral blood, bone marrow and liver tissues were less than 1@1000, but local human cell frequencies of 207.92@1000 and 392.41@1000 were detected in the liver tissues of 2 transplanted goats. CONCLUSIONS: The existence and long-term survival of human cells in transplanted goats detected by FISH indicated that goats were appropriate recipients for hHSC in utero transplantation. The lower human cell frequencies in blood and bone marrow, and the higher local human cell frequencies in liver tissues suggested that the microenvironment of goat liver tissues might favor the survival, proliferation and differentiation of human cells. PMID- 12905703 TI - [Cloning, expression, and alternative splicing of the novel isoform of hTCP11 gene]. AB - OBJECTIVE: To identify a novel isoform of hTCP11 gene and investigate its expression and alternative splicing. METHODS: According to the sequence of human ESTs which are highly homologous to hTCP11a, primers for PCR were synthesized. Then, the amplified fragments were cloned and sequenced; some methods including BLAST, ClustalW and RT-PCR were used for genomic analysis, study of alternative splicing and gene expression among multiple tissues and different testis tissues. RESULTS: A novel isoform of hTCP11 gene was isolated. It encodes a 440 amino acid protein that is highly homologous to the mouse 566 amino acid protein which is important to sperm function because it encodes the receptor for fertilization promoting peptide (FPP). Among TCP11a, TCP11b and TCP11c, the complicated alternative splicing was found. RT-PCR analysis of RNA extracted from human tissues revealed that the gene is only expressed in fertile adult testes, but not in azoospermic patient testes, fetal testes or other human tissues. CONCLUSION: Our results along with the mouse Tcp-11 function suggest that the isoforms of TCP11 gene play important roles in sperm function and fertility. PMID- 12905705 TI - [Mutation-function analysis in the lipoprotein lipase gene of Chinese patients with hypertriglyceridemic type 2 diabetes]. AB - OBJECTIVE: To investigate the role of lipoprotein lipase (LPL) gene on Chinese patients with hypertriglyceridemic type 2 diabetes. METHODS: Three subject groups, including hypertriglyceridemic group, normalipidemic type 2 diabetes group and healthy controls, were recruited and screened for sequence changes in LPL gene with PCR, SSCP, restriction analysis and direct DNA sequencing. LPL mass and activity in post-heparin plasma and in in vitro expression were investigated. Comparative modeling was performed via Swiss-PDB Viewer to provide the potential 2-D structures of wildtype and mutant proteins. RESULTS: Four missense mutations, Ala71Thr, Val18Ile, Gly188Glu and Glu242Lys, were identified in patients with hypertriglyceridemic type 2 diabetes, and not in both normalipidemic diabetes and the control subjects. The four missense mutations were located in the highly conserved amino acid sites, which are involved in highly conserved exon 3, 5, or 6 regions. They led to reduced LPL mass and enzyme activities in both post heparin plasma and in vitro expression. The modeled structures displayed the differences to a great extent between the mutant and wide-type molecules. CONCLUSION: These results indicated that the 4 missense mutations lead to LPL deficiency and subsequent hypertriglyceridemia. The LPL deficiency predispose a progressive diabetic pathway to those affected individuals. LPL gene is one of susceptibility gene for hypertriglyceridemic type 2 diabetes. PMID- 12905706 TI - [Ten novel mutations in the phenylalanine hydroxylase gene identified in Chinese patients with phenylketonuria]. AB - OBJECTIVE: To study the molecular basis of the phenylalanine hydroxylase (PAH) gene mutation in Chinese patients with phenylketonuria (PKU). METHODS: Using PCR/SSCP and DNA sequencing, we studied the mutations in exons 3, 5, 7, 10, 11, 12 of PAH gene. Totally 120 unrelated children with PKU and their parents from the northern region of China were included in the analysis. RESULT: Ten novel mutations were first time identified in Chinese PKU population as I65T, S70del, G239D, R241fsdelG, L255S, P281L, G346R, L367fsinsC, R400S and Ivsllnt2t-->c. The mutations G239D, R241fsdelG, R400S and Ivsllnt2t-->c have not been yet described in International PAH. In the present study we firstly identified the deletion, insertion and frameshift mutations of PAH gene in China PKU population. So far the mutant type of PAH gene in Chinese included: missense, nonsense, splice, silence, deletion, insertion and frameshift. Novel mutations mainly existed in exon 7: four in exon 7, two in exon 3, two in exon 11, one in exon 10 and one in intron 11. Each proportion of the ten novel mutations was very low (0.42%-1.3%). CONCLUSION: This study demonstrated the high heterogeneity of the PAH gene and the variety of the mutant type of Chinese PKU population and confirmed the exon 7 was the hot spot of PAH gene mutation. PMID- 12905707 TI - [Correlation analysis between WNK4 gene and essential hypertension]. AB - OBJECTIVE: To investigate association of mutation in WNK4 gene with essential hypertension and to analyze the expression of WNK4 gene. METHODS: cSNP in WNK4 gene in a small samples was detected by sequencing, then PCR-RFLP was performed in 98 patients with essential hypertension and 95 control subjects. The expression profile of WNK4 gene was tested by RT-PCR. RESULTS: A cSNP was detected in WNK4 gene exon7 G1662A, and there were significant differences in the distribution of allele frequency of G1662A between essential hypertension group and control group. WNK4 gene were expressed in the tissues of kidney, brain, lung, heart, spleen and intestine of fetus. CONCLUSION: WNK4 gene is well correlated with essential hypertension. PMID- 12905708 TI - [Overexpression of p21WAF1 and p53 in human lung adenocarcinoma cell line]. AB - OBJECTIVE: To study the growth inhibitory effects of p21WAF1 and p53 overexpression in human lung adenocarcinoma cell line. METHODS: The p21WAF1 and p53 gene were transfected respectively into a human lung adenocarcinoma cell line, GLC-82. Flow cytometry (FLC), transmission electron microscopy (EM) and TUNEL technique were used to evaluate cell growth and identify apoptosis. RESULTS: The GLC-82 transfected by p21 plasmid showed increased cell number in G1 phase of cell cycle, decreased proliferation potential and decreased cloning efficiency. Apoptosis have not been detected neither on EM nor by TUNEL technique, whereas the GLC-82 infected by Ad-p53 showed significantly decreased proliferation potential and some of them even died, in addition apoptosis was confirmed by TUNEL technique. CONCLUSION: The results indicate that p21WAF1 and p53 can inhibit proliferation; p53 also can induce apoptosis of lung adenocarcinoma cell. Therefore, these two genes should have a wide application in gene therapy of tumors in future. PMID- 12905709 TI - [Detection of mtDNA 11778 (G-->A) point mutation in a family with Leber's hereditary optic neuropathy by site-specific polymerase chain reaction]. AB - OBJECTIVE: To find a simple, fast, accurate, and quantitative PCR-based method for mutation detection, so as to identify mitochondrial DNA 11778 G-->A point mutation in patients with Leber's hereditary optic neuropathy (LHON). METHOD: On the basis of sequencing of mtDNA from LHON proband, M primer for mutation and N primer for normal were designed to be coupled with reverse primer respectively. Specific PCRs were done on an amplifying condition with high stringency such as a well controlled annealing temperature, low Mg2+ concentration and less thermal cycles. The objective pedigree includes 10 individuals, were against 40 normal control persons. RESULTS: Different ratios of indicative mtDNA 11778A-->G mutation were checked out from the proband, affected maternal members and a 10 year-old boy (up to now no appearance yet), whereas not appeared on normal spouses, paternal offsprings in the family, neither did on 40 controls. CONCLUSION: This site-specific PCR method is a kind of general mutation analysis way, without the restriction of existence of endonuclease site. It can be applied for the gene diagnosis of known-mutation hereditary diseases such as LHON. PMID- 12905710 TI - [Screening by maternal serum markers for Down's syndrome]. AB - OBJECTIVE: To investigate the optimal method of screening for Down's syndrome (DS) with maternal serum mankers. METHODS: Screening by maternal serum markers for Down's syndrome was offered to all 2886 pregnant women in Peking Union Medical Hospital during 1996.11-2001.3. Alpha-fetoprotein (AFP), human chorionic gonadotrophin (free beta-HCG) were used as markers during the first year of pregnancy. Alpha-fetoprotein, free human chorionic gonadotrophin (HCG) and pregnancy-associated plasma protein A (PAPP-A) were used as mid pregnancy and first-trimester markers in next three years. Amniocentesis and (CVS) were done in those defined as risk cases. RESULTS: The detection rate of Down's syndrome by maternal serum markers was 3.8% (11/2886). The proportion of false positive results in group of triple markers (alpha FP, free beta-HCG, PAPP-A) was 5%. CONCLUSIONS: The PAPP-A was a good marker to detect Down's syndrome in early pregnancy and may be used to predict the outcome during mid trimester of pregnancy. The AFP and free beta-HCG can be useful markers to detect Down's syndrome and fetal abnormality. While prenatal diagnostics can be shifted to an early pregnant period. PMID- 12905711 TI - [Bone marrow stem cells transplantation improve locomotive function of dystrophin/utrophin gene double knock-out mice]. AB - OBJECTIVE: Study the improvement of locomotive faculty of dystrophin/utropin gene double knock-out mice (dko mice) by transplanting bone marrow stem cells. METHODS: The bone marrow stem cells of C57BL/6 mice (4- to 5-weeks age) were cultured in vitro for three days, before transplanted intravenously (1.0 x 10(7) for each) into 11 dko mice (7- to 8-weeks age). The dko mice were irridiated with 7Gy gamma-ray before transplantation. 8-9 weeks after transplantation, the locomotroy function, electromyography items and expression of dystrophin in transplanted mice and controls were observed. RESULTS: 8-9 weeks after transplantation, the dropping times of hauling wire were 3.09 +/- 2.47, compared with that of the control dko mice(16.78 +/- 3.60), there are distinct differences. About electromyography items, the duration of active potential and amplitude of maxim contractions were (4.99 +/- 1.62) ms and(2872 +/- 1474.33) microV, compare with those of control dko mice(3.69 +/- 0.40) ms and(1210.0 +/- 551.0) microV, respectively, about 7% fibers of the muscle tissue of transplanted dko mice expressed dystrophin protein. CONCLUSIONS: 8-9 weeks after transplanted with homology bone marrow stem cells, the locomotive function and electromyography items of transplanted dko mice were obviously improved, and about 7% muscle tissue fibers of the mice expressing dystrophin protein were observed. It suggested that there is an ideal prospect for DMD therapy with bone marrow stem cells transplantation. PMID- 12905712 TI - [Dual regulation by delta opioid receptor agonists on the delayed rectified potassium channels in NG108-15 cells]. AB - OBJECTIVE: To investigate the dual effects by the delta opioid receptor agonists DPDPE on the delayed rectified potassium channels in NG108-15 cells. METHODS: A series of outward currents were evoked in NG108-15 cells by depolarizing voltage from -50 mV to +80 mV at holding potential of -90 mV. These currents were delayed rectified potassium currents. Relatively selected delta opioid receptor agonists DPDPE of higher and lower concentrations were used to modulate the delayed rectified K+ current in NG108-15 cells. Opioid receptor antagonist Naloxone (NAL) and relatively selected delta opioid receptor antagonist Naltrindole (NTI) were used in the present experiments for the characterization of the actions of opioid receptors. RESULTS: The relatively higher concentrations of delta opioid receptor agonist DPDPE (> or = 10(-6) mol/L) significantly increased the amplitude of the delayed rectified K+ current. On the contrary, the relatively lower concentrations of DPDPE (< or = 10(-12) mol/L) decreased the amplitude of the delayed rectified K+ current (P < 0.05). Furthermore both the increase and decrease were time-dependent. CONCLUSIONS: delta opioid receptor agonist has dual regulatory effects on the delayed rectified potassium channels in NG108-15 cells. PMID- 12905713 TI - [Clinical application of presurgical embolization of meningiomas]. AB - OBJECTIVE: To evaluate the clinical efficacy and safety of presurgical embolization of meningiomas. METHODS: Matched analysis of samples of embolized and nonembolized groups of meningiomas were observed for variables of clinical efficacy in estimated blood loss, number of transfusions and length of post operative recovery. RESULTS: All mean value of variables of embolized group, were lower than that of non-embolized (estimated blood loss, 1402.97 +/- 1171.60 ml versus 1852.94 +/- 993.40 ml; number of transfusions, 1325.53 +/- 1040.15 ml versus 1747.06 +/- 959.24 ml; length of post surgical recovery, 30.18 +/- 20.24 days versus 33.29 +/- 27.54 days); however, only the estimated blood loss and number of transfusions-variables were significant. There were no major complications caused by the embolization procedure. CONCLUSION: Endovascular devascularization of meningiomas is beneficial for meningiomas because it diminishes the necessity of intra operative transfusion and decreases blood loss. Otherwise the embolization procedure is safe. PMID- 12905714 TI - [A clinical analysis of 103 cases of adrenal incidentaloma]. AB - OBJECTIVE: To improve the diagnosis and treatment of adrenal incidentaloma (AI). METHODS: Retrospective analysis of 103 AI patients in PUMC Hospital from 1994 to 2002. RESULTS: Of the 103 cases (43 men and 60 women), most patients were 40-60 years old. Eighty-five cases (82.5%) were benign masses, of which 36 (35.0%) were non-functional adrenal cortical adenoma, and 12 (11.7%) were pheochromocytoma, 1 (1%) was Cushing's syndrome and 1 for aldosteronomas. Three cases did not show clinical feature and the diagnosis of pheochromocytoma were based on 131I-MIBG. Among 18 (17.5%) malignant tumors, adrenal carcinomas account for the majority (8 cases, 7.8%). Patients with malignant masses were significantly younger than patients with adenomas (P < 0.05), and the adenomas were significantly smaller than malignancy (P < 0.001). A cut-off at 4.0 cm of tumor mass size was set for differentiating benign and malignant tumors. CONCLUSIONS: During the diagnosis of AI, it's essential to evaluate hormonal activity and assess for the risk of malignancy. If the mass size is 4 cm or larger, the risk of malignancy increases. More attention should be paid to subclinical hyperfunctional state. PMID- 12905715 TI - [Molecular cloning of Plasmodium yoelii dynamin-like protein (PyDyn) gene and the immunological character of its domains]. AB - OBJECTIVE: To identify and clone a new full ORF gene of PyDyn (Plasmodium yoelii dynamin-like protein), and examine the protection of their expression products. METHOD: Using the P. yoelii Genome technology and RT-PCR. RESULTS: The full ORF gene of PyDyn was amplified from mRNA of the erythrocytic stage of P. yoelii., three domains of PyDyn were expressed in E. coli., and the fairly positive immunogenicity of them was showed by IFA. The full ORF gene of PyDyn was 2,433 bp and encode 811 amino acids. Its Gene Bank access number is AF458071. PyDyn belongs to the dynamin-like protein family according to its property. CONCLUSION: The new full ORF gene of PyDyn is obtained and identified; their expressed domains are probably new candidates for malaria vaccine. PMID- 12905716 TI - [Gene clone and its characteristics on band 7-like protein in Plasmodium falciparum FCC1/HN]. AB - OBJECTIVE: To identify and clone the gene named pfstom gene which encoding the protein belonging to band 7 family and to do primary research on its function. METHODS: Based on the finished data in international public malaria database, coding sequence of pfstom cDNA was obtained by RT-PCR from FCC1/HN. Its phylogenetic profiles and the homogeny were analyzed by some softwares. After Prokaryotic expression, C terminal of Pfstom protein was expressed by Pet30a system. Recombinant Pfstom protein was used to immol/Lunize rabbit and then serum was harvested and the IgG was purified for Western blot. RESULTS: The coding sequence of pfstom is 1,125 bp which encoding 374 amino acids with C-terminal fragment being homogenous to stomatin-like protein which belongs to band 7 family. Phylogenetic profiles analysis revealed its homogeny to stomatin. Western blot showed its stage-specific expression in trophozoite. CONCLUSION: Pfstom belongs to band-7 family. It was expressed specifically in trophozoite in erythrocyte stage of plasmodium falciparum. It was not expressed in ring stage. And it is membrane-related protein. All these results provided the foundation for further research on pfstom. PMID- 12905717 TI - [Gene therapy of rat prolactinomas mediated by adenoviral vectors with rat tyrosine hydroxylase gene]. AB - OBJECTIVE: To investigate the potential of gene therapy of rat prolactinomas mediated by adenoviral vectors with a gene encoding rat tyrosine hydroxylase. METHODS: Recombinant replication-deficient adenovirus named Ad-GFP-TH with rat TH cDNA and control adenovirus named Ad-GFP were constructed by homologous recombination in bacterial cells. The rat pituitary prolactinoma cell line MMQ are chosen as the target cells to study the effect of gene therapy on their growth and prolactin secretion mediated by Ad-GFP-TH. RESULTS: Recombinant Ad-GFP TH and Ad-GFP were successfully reconstructed. Transfection of MMQ cells with Ad GFP-TH not only restrained their growth but also decreased their PRL secretion. CONCLUSION: Gene therapy may serve for a potential treatment for prolactinomas, especially invasive prolactinomas. PMID- 12905719 TI - [Effect on dose owing to source displacement in coronary endovascular brachytherapy]. AB - OBJECTIVE: To investigate the effect on the dose distribution of source position displacement with the target region margin in catheter-based 192Ir line source endovascular brachytherapy. METHOD: Dose rate distribution along longitudinal axes was estimated by the formula recommended by AAPM No.60 and No. 43. RESULTS: In the two cases of source displacement (1.1 and 5 mm) doses of target region margin was decreased dramatically (at most 53.9% and 565.8% respectively) were compared to that of no source displacement, and the affected range was 6 mm and 9 mm respectively. CONCLUSION: Source displacement will lead to the decrease of dose in target region margin. PMID- 12905718 TI - [Diagnosis and treatment of mesenteric venous thrombosis: analysis of eleven cases]. AB - OBJECTIVE: To evaluate the diagnosis and treatment of mesenteric venous thrombosis. METHODS: The clinical data of 11 cases diagnosed as mesenteric venous thrombosis between 1992 and 2001 in PUMC Hospital were analyzed retrospectively. RESULTS: Postoperative state(27.3%), especially cirrhosis and portal hypertension, and other history of thrombosis (27.3%) were the most common causes. Thrombolysis was performed successfully in two of the eleven cases. The rest of them were misdiagnosed in other hospitals and operated. No patient died after operation, and one (11.1%) recurrence was found. CONCLUSIONS: Early application of anticoagulant is necessary for patients with thrombosis risks. For suspected patients, early computed tomography (CT) and DSA examination should be performed, and prompt thrombolysis and anticoagulation therapy can be performed to avoid the bowel resection after definite diagnosis. To reduce the recurrence, anticoagulant should be maintained for a proper time. PMID- 12905720 TI - [Detection of esophageal intubation-assessment of several methods in clinical anesthesia]. AB - OBJECTIVE: To evaluate the sensitivity and reliability of several widely used tests for prompt detection of inadvertent esophageal intubation. METHODS: Both endotracheal and esophageal intubations were made on 40 adult patients undergoing general anesthesia. The tests such as auscultation of bilateral apex of lungs and epigastrium by inexperienced examiners, capnography, SpO2, chest and upper abdomen movements, and airway resistance were evaluated. RESULTS: 90% and 96.25% cases in esophageal intubation were correctly diagnosed via auscultation of bilateral apex of lungs or epigastrium respectively. During esophageal ventilation, abdominal distension was found in 87.5% of cases, but none of them showed chest movements. Meanwhile, PetCO2 fluctuated between 1-2 mmHg, in association with a quick decline of SpO2 in 156 +/- 11 seconds. The airway mean resistance increased, whereas the period of plateau decreased significantly. CONCLUSIONS: (1) Auscultation of epigastrium in combination with bilateral apex of lungs is recommended because of the improved accuracy in tube positioning. (2) Capnography is the most reliable technique for the prompt detection of esophageal intubation, whereas other parameters do not seem to be of comparable value. PMID- 12905721 TI - [Transvaginal ultrasound assessment of physiological changes of Chinese women's cervix]. AB - OBJECTIVE: To observe Chinese women's cervical physiological changes by transvaginal ultrasound. METHODS: 637 normal female volunteers were studied. Cervical length, width and canal width were measured by transvaginal ultrasound. Cervical length multiplied by width was the biggest vertical section area of cervix(cervical area). RESULTS: The cervical length, width, area and canal width in the menopausal women were significant smaller than that in the un-menopausal women (P < 0.0001). Multinomial logistic regression showed that cervical changes were mainly affected by menopause year, gestation and para. The cervical length, width, area and canal width were negative correlated with menopause year significantly. The cervical and canal width were positive correlated with gestation. The regressive equation was Cervical area (cm2) = 3.69 + 0.23 Gastation -0.16 Menopause year. CONCLUSIONS: Cervical physiological changes are mainly affected by the menopause year, gestation and para. The menopause year is the most important factor in the cervical physiological changes. PMID- 12905722 TI - [Somatostatin mRNA expression in hippocampus of diabetic rats model]. AB - OBJECTIVE: To investigate the relationship between diabetes and somatostatin (SOM). METHODS: We established a streptozocin-induced diabetic rats model and studied the changes of SOM mRNA in hippocampi of diabetic and normal rats by using in situ hybridication and computer image analysis. We studied the mechanism of chronic diabetes encephalopathy by observing the changes of somatostatin mRNA in the hippocampus of diabetic rats was studied. RESULT: The number, light density and average area of positive cells in dentate gyrus area of hippocampus in diabetes model were reduced significantly as compared with normal rats (P < 0.01). CONCLUSION: A decline in SOM mRNA expression may play a role in chronic diabetes encephalopathy because of SOM effect in brain. PMID- 12905723 TI - [Neuroelectrophysiological basis and surgical treatment of essential tremor]. AB - OBJECTIVE: To investigate the relationship between limb tremor and neuronal firing in thalamus (Vim) and retrospectively review the clinical effects and safety of the surgical treatment of essential tremor (ET). METHODS: Forty-two ET patients received microelectrode-guided thalamotomy and 11 cases were quantitatively evaluated with FAHN rating scales pre- and post-operatively. RESULTS: There were electrophysiological tremor-related neurons in ventrolateral part of thalamus. Lesioning of those neurons abolished contralateral limb tremor in all of the patients. No permanent contralateral weakness, dysarthria and hemorrhage were observed. CONCLUSION: "Tremor cell" in thalamus plays a key role in the symptom of ET patients. Destruction of those cells may completely and permanently abolish tremor symptom. PMID- 12905724 TI - [A clinical analysis of glucagonoma syndrome (four cases reported)]. AB - OBJECTIVE: To study the clinical features, diagnosis, and treatment of glucagonoma syndrome (GS) for providing clues for the recognition of this disorder in clinical practice. METHOD: The clinical and laboratory findings of four confirmed patients with GS were analyzed retrospectively. RESULTS: All four patients had typical clinical manifestations of necrolytic migratory erythema (NME), elevated glucagon level in serum and hepatic metastasis. The skin rashes disappeared gradually and serum glucagon level decreased after operation and somatostatin treatment. CONCLUSIONS: NME is the most specific clinical finding of GS. Biopsy of the lesions, glucagon level in serum and various radiological examinations should be done in order to confirm the diagnosis. Surgical resection, chemotherapy, and somatostatin are the main therapies of GS. PMID- 12905725 TI - [Treatment and clinicopathologic analysis of mucosa-associated lymphoid tissue lymphoma of the salivary glands]. AB - OBJECTIVE: To further understanding of lymphoma of salivary gland through clinicopathologic analysis. METHODS: Clinical findings, pathologic features, clinical staging, therapy and prognosis of 4 cases were reviewed and clinically analysed. RESULTS: Different treatment were received by the 4 patients, one had stage IIIA disease and three had stage IE disease. All patients got their illness completely remitted. CONCLUSION: Mucosa-associated lymphoid tissue lymphoma of the salivary glands is an indolent disease. Different treatments can all result in prolonged remission, and it has better outcome than other NHL. PMID- 12905726 TI - [An eighteen-year study on phenylketonuria]. AB - A study on phenylketonuria (PKU) has been carried out in China-Japan Friendship Hospital since 1984. The results revealed that: (1) Totally 603 patients with PKU were diagnosed and treated in the hospital from October 1984 to September 2002. Among which 136 cases were identified by neonatal screening and treated within 3 months. One hundred and ninety-five cases were treated when the children were 3 12 months of age. Another 272 PKU children were diagnosed when they were more than 1 year old. All of these late-treated cases had some signs and symptoms of PKU. Mental retardation was found in 467 cases and various patterns of seizures in 119 cases. After treatment with low-phenylalanine diet, the follow-up for early-treated patients revealed that their physical and mental developments were normal. In late-treated patients, abnormal behaviour was significantly improved and their developmental quotient were elevated. Prenatal gene diagnosis of PKU risk foetus in 22 PKU families was successfully performed. (2) Urinary pterins obtained from 369 HPA patients were measured by HPLC. Twenty two patients with BH4 deficiency have been recognized. Six single base mutations were detected in 18 unrelated northern Chinese BH4 deficiency families, and the mutations at nucleotides 259C-->T and 286G-->A were common mutations. Eighteen BH4 deficient patients were treated with BH4, L-dopa and 5-hydroxytryptophan, and the results were satisfactory. (3) The abnormal rate of EEG was high in untreated patients with PKU, mainly showing epileptiform discharges and partly showing background activity abnormality. The most frequent finding was patchy areas of increased signal intensity in white matter on MRI in the brain of PKU patients, while delayed myelination and brain agenesis were often detected. After dietary treatment, follow-ups with EEG and MRI revealed that the abnormalities were decreased significantly. (4) The relationship between genotype and intellectual phenotype was examined in 29 late-treated patients with classical PKU. It was found that the genotype of 22 patients were compatible with intellectual phenotype and not well matched in 7 cases. The result indicate that the genotype was well matched with intellectual phenotype in classical PKU patients. PMID- 12905727 TI - [Progress in the studies on cytomegalovirus retinitis]. AB - Cytomegalovirus retinitis (CMVR) is the commonest opportunistic ocular infection in patients with human immunodeficiency virus (HIV) infection/acquired immunodeficiency syndrome (AIDS), typically occurs when CD4+ T cell counts fall below 50/mm3. CMVR accounts for the majority of the vision loss associated with HIV-related eye diseases. However progress in the studies on CMVR, including the prevalence, clinical features, differential diagnosis and recent advances in the management of CMVR is reviewed. PMID- 12905728 TI - [IL-15 and autoimmune disease]. AB - This review attempts to define the relationship between IL-15 and autoimmune disease (AID) from IL-15's tissue distribution, expression regulation and biologic function. Meanwhile, five IL-15 dysregulation-related AIDs and four IL 15/IL-15R-targeted AID therapies are described. PMID- 12905729 TI - [Role of Ski/SnoN protein in the regulation of TGF-beta signal pathway]. AB - TGF-beta signal pathway plays an important role in the cell growth, differentiation, formation of extracellular matrix, embryo development and carcinogenesis, etc. However, the regulation of TGF-beta pathway is not totally understood. In 1999, three independent research groups found that Ski/SnoN protein could inhibit the TGF-beta mediated transcription by recruiting N-CoR, a transcription co-repressor. Later studies suggested that TGF-beta and SMADs degraded the Ski/SnoN protein by mediating ubiquitin linkage, showing negative feedback regulation. The important findings in Ski/SnoN laid the theoretical foundation for demonstrating the function of TGF-beta signal pathway. PMID- 12905730 TI - [Steroids hormone and bone]. AB - Bone is an active tissue in which the processes of remodeling are continuous to ensure normal bone integrity and strength. Steroids play an important role in regulating bone growth, development and remodeling. Glucocorticoids excess will induce bone damages especially osteoporosis. Otherwise, estrogen and androgen are bone protective steroids in both female and male. To develop a new selective steroid receptor modulator is one of the targets in future study to treat osteoporosis. PMID- 12905731 TI - [Some problems of postmenopausal osteoporosis prevention with hormone replacement therapy]. AB - Estrogen deficiency is the one of pathogenetic causes of postmenopausal osteoporosis. Exogenous estrogen may prevent postmenopausal bone loss, so to prevent postmenopausal osteoporosis and osteoporotic fracture. The effect of estrogen on postmenopausal syndrome can not be substituted with other medicine. However, the attention should be paid to the safety of hormone replacement therapy (HRT) during its long term use, and HRT should be used individually. PMID- 12905732 TI - [Effect of aging on the ability of growth and differentiation of rat bone marrow stromal cells]. AB - OBJECTIVE: To measure the osteogenesis and adipogenesis potentiality of rat marrow stromal cells (MSCs) derived from 3, 6, 9, 12-month-old doner rats. METHODS: Rat MSCs were induced to osteoblast or adipocyte by osteogenic inducer or adipogenic inducer. In different times, 3 and 12-month-old rat MSCs were observed by histochemistry staining; the mRNA level of type I collagen and lipoprotein lipase of 3, 6, 9, 12-month-old rat MSCs were measured by RT-PCR. RESULTS: The expression of alkaline phosphatase (ALP) of the control group and induced group of 12-month old rat MSCs was less than that of 3-month old rat MSCs after 1-week osteogenic induction. 12 days later, calcification was observed in 3 month old group. Lipid droplets occurred in the cells of 12-month old group after 2-day adipogenic induction, while these droplets occurred after 3-day or 4-day induction in 3-month old group. The mRNA level of type I collagen decreased with the increase of age. The mRNA level of lipoprotein lipase of younger rats was lower than that of older rats. Both of the changes were more significantly with the increase of the induction time. CONCLUSIONS: With the increase of age, the ability of osteogenesis of rat MSCs decreased, but the ability of adipogenesis increased. PMID- 12905733 TI - [Relationship between body composition and bone mineral density in healthy premenopausal women]. AB - OBJECTIVE: To investigate the relative contribution of fat mass and lean mass on bone mineral density (BMD) in premenopausal healthy women. METHODS: The BMD at lumbar spine, proximal femur and total body, together with fat mass and lean mass was determined by dual-energy X-ray absorptiometry (DEXA), the body height, weight, waist, and hip circumference were also measured, and body mass index (BMI) and waist-hip ratio were calculated in 282 premenopausal women. RESULTS: Fat mass was a major determinant for BMI, BMI and lean mass were positively related with L2-4, proximal femur and total body BMD (P = 0.000 for all), and lean mass were the only independent factor contributing to L2-4 (standardized coefficient beta = 0.282, P = 0.000), proximal femur (beta = 0.336, P = 0.000) and total body BMD (beta = 0.361, P = 0.000) in stepwise regression analysis. The relationship between BMI and BMD was further improved after controlling fat mass, while decreased or even lost when controlling lean mass. CONCLUSIONS: Lean mass was an important factor determining BMD in premenopausal women. PMID- 12905734 TI - [Association of Apa I polymorphism of vitamin D receptor gene with bone mass in men]. AB - OBJECTIVE: To investigate the association of Apa I polymorphism in vitamin D receptor (VDR) gene with bone mass in men. METHODS: The VDR Apa I genotype was determined by PCR-restriction fragment length polymorphism (RFLP) in 388 unrelated healthy men aged 46-80 years of Han nationality in Shanghai city. Bone mineral density (BMD) and bone mineral content (BMC) at lumber spine 1-4 (L1-4) and at any sites of proximal femur including to femoral neck (Neck), trochanter (Troch) and Ward's striangle (Ward's) were measured by duel-energy X-ray absorptiometry. RESULTS: Frequencies distribution of VDR Apa I genotype were aa for 48.1%, Aa for 44.2% and AA 7.7%. The allele frequencies of Apa I polymorphism were in Hardy-Weinberg equilibrium. No significant association was found between Apa I genotype and BMD or BMC in group of all population or in subgroup of men below 60 years. In men above 60 years, the significant association was found between VDR Apa I genotype and BMD or BMC at L1-4, Neck and Ward's (P < 0.05, P < 0.01) and compared with Aa and aa genotype, AA genotype had significantly higher mean BMD and BMC at L1-4, Neck and Ward's (P < 0.05, P < 0.01). But Apa I genotype is not associated with BMD and BMC at Troch. CONCLUSIONS: Apa I polymorphism is associated with bone mass in men above 60 years, and AA genotype has higher bone mass. Apa I polymorphism in VDR gene possibly influence loss of trabecular and cortical bone mass in old men. PMID- 12905735 TI - [Association of calcitonin receptor gene polymorphism with bone mineral density in Shanghai women]. AB - OBJECTIVE: To investigate the association of calcitonin receptor (CTR) gene polymorphism with bone mineral density (BMD) in premenopausal and postmenopausal women. METHODS: CTR genotypes were determined by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) in 184 premenopausal women and 199 postmenopausal women in Shanghai area. BMD at lumbar spine (L2-4) and femoral neck (FN) were measured by dual-energy X-ray absorptiometry (DEXA). RESULTS: The distribution of CTR genotypes in 383 Shanghai women were CC genotype 83.8%, TC genotype 14.6%, TT genotype 1.6%, respectively. BMD at FN of CC genotype was significantly higher than TC and TT genotypes (P < 0.01) in postmenopausal women. But there was no difference in BMD of different CTR genotypes in premenopausal women. Multiple regression analysis showed that CTR genotypes were associated with FN BMD in postmenopausal women (P < 0.05). CONCLUSIONS: The polymorphism of CTR gene was associated with BMD in postmenopausal women. PMID- 12905736 TI - [Manufacture and evaluation of a spine phantom used in the cross-calibration of DXA bone densitometry]. AB - OBJECTIVE: To design and manufacture a reliable spine phantom used in the cross calibration and quality control of dual X-ray absorptiometry (DXA). METHODS: A hydroxyapatite quality control phantom was designed and made through three steps: solid water, bone phantom material and integration, then evaluated the phantom on four different types of DXA machines made by LUNAR company. RESULTS: Among the four DXA densitometers, the Expert fan beam densitometer had the biggest accuracy errors of bone mineral density (BMD), bone mineral content(BMC) and area values while the other three one narrow fan beam (Prodigy) and two pencil beam densitometers-had small errors. Of the three indexes measured by all the machines, BMD error was the smallest (-15.4%-11.5%), with the Prodigy's BMD was most outstanding. BMD errors at the higher density ends were small, tend to be positive values while the errors at the lower ends were big, tend to be negative. In cross calibration, giving consideration to the differences between the both ends, it is better to use the regression equation to correct. The base line of precision error derived by scanning the phantom once a day for consecutive 25 days was better than that derived by scanning the phantom 25 times consecutively on the same day. As to precision error, the coefficient of variation (CV) of scanning-25 times-a day was the smallest (0.0043) while the CV of 12 adults measurements was the biggest (0.0078). CONCLUSIONS: This phantom can be used in the quality control and cross-calibration of different types of DXA machines. PMID- 12905737 TI - [Effect of femoral rotation on hip bone mineral density measurement]. AB - OBJECTIVE: To measure hip bone mineral density (BMD) on different femoral rotations and to identify the influence of femoral rotation during hip densitometry. METHODS: Forty women were recruited at the out-department of orthopedic, with the average age of 60.7 years (45-85 years), the average height of 161.8 cm (150-175 cm), and the average weight of 67.4 kg (48-80 kg). BMD measurement of hip was done in neutral position (0 degrees), 15 degrees and 30 degrees of internal rotation under quality control of DXA machine. All the subjects were asked to get down scan table and reposition after each measuring in different rotation. RESULTS: Hip BMD of proximal femur (femoral neck, Ward's area and trochanter region) have significant variations in measurement (P < 0.01) among neutral position, 15 degrees and 30 degrees of internal rotation. There were more BMD change at the cancellous region (Ward's triangle and trochanter region) than at cortical bone area (femoral neck region). Statistical analysis showed a significant positive correlation for hip BMD change between neutral degrees to 15 degrees of internal rotation and 15 degrees to 30 degrees of internal rotation. But no correlation between neutral degrees to 30 degrees of internal rotation for hip BMD change. CONCLUSIONS: Femoral rotation was shown to have a significant effect on hip BMD measurements. Cortical bone region of hip have a less BMD change than cancellous bone region. Proper positioning of the femur during scan can improve precision significantly. PMID- 12905738 TI - [Changes of urinary deoxypyridinoline crosslink/creatinine in rats after ovariectomy and anti-osteoporotic intervention]. AB - OBJECTIVE: To observe the changes of urinary deoxypyridinoline crosslink/creatinine (UDpd/Cr) in rats after OVX and intervention by estrogen and bisphosphonate and investigate the possible application of deoxypyridinoline in osteoporosis diagnosis and treatment. METHODS: 40 female 6-month-old virginal Wistar rats were divided into 5 groups, ovariectomized or sham ovariectomized. (1) Ovxb (n = 8): sacrificed at 6 weeks after OVX; (2) Sham (n = 8): sham ovariectomized; (3) Ovxe (n = 8): sacrificed at 14 weeks after OVX; (4) O + E (n = 9):OVX + 17 beta estradiol [20 micrograms/(kg.d) ih]; (5) O + C (n = 7):OVX + cimadronate [0.2 mg/(kg.d)]; Treatment started 6 weeks after OVX and lasted 8 weeks. Rats in group 2-5 were sacrificed at 14 weeks after OVX. Urinary and serum biochemical parameters were measured, pQCT scanning of femur, bone biomechanical test in femur were determined. RESULTS: OVX resulted in increasing of UDpd/Cr 133.3% (P < 0.01). The ratio of UCa/Cr also increased in OVX groups but without any significant compared with Sham (P > 0.05). UDpd/Cr were reduced by 54.6% and 51.8% (P < 0.01) in O + E, O + C group respectively compared with Ovxe. The significant negative correlationships were found between UDpd/Cr and bone mass, BMD and biomechanic characteristics. CONCLUSIONS: UDpd/Cr ratio is a sensitive bone resorption marker, a marked changes were observed when the rats ovariectomized or treated with estradiol and cimadronate. There were best correlation between UDpd/Cr and bone mineral density and bone biomechanic characteristics. It is fair to apply UDpd/Cr ratio for osteoporosis diagnosis and treatment. PMID- 12905739 TI - [Therapeutic effect of recombinant human parathyroid hormone(1-34) on osteoporosis of ovariectomized rats]. AB - OBJECTIVE: To investigate the therapeutic effect of recombinant human parathyroid hormone(1-34) [rhPTH(1-34)] on osteoporosis of ovariectomized rats. METHODS: The model of osteoporosis was formed after 3 months of ovariectomy with 6-month age of 80 rats. Another 20 rats was control of sham operation. rhPTH(1-34) was subcutaneously injected once daily with 5, 10, 20, 40 micrograms/kg for 3 months. There were 10 rats in each group. The control of therapy included Salmon Calcitonin to 10 rats and Alendronate sodium to 10 rats. The bone weight of dry and ash, bone mineral density, bone biomechanical property, trabecular area, bone mineral deposition and serum alkaline phosphatase, Ca, P and urinary Pyridinoline/creatin (Pyd/Cr) were measured after the end of therapy. RESULTS: When administered to animals as a single subcutaneous injection once daily, rhPTH(1-34) increased obviously bone mass, bone biomechanical property and trabecular area, as well as bone deposition compared with the animals of control group. The bone architecture was ultimately improved by rhPTH(1-34) therapy. CONCLUSIONS: Rats of ovariectomized-induced osteoporosis possess obvious effect of treatment with low dose of rhPTH(1-34) administered once daily. PMID- 12905740 TI - [Assessment of preoperative localization techniques for patients with primary hyperparathyroidism]. AB - OBJECTIVE: To evaluate the sensitivity and usefulness of 99mTc-sestamibi scintigraphy (SS) and neck ultrasonography (US) as preoperative localization procedures in patients with primary hyperparathyroidism (pHPT). METHODS: 160 patients with proved pHPT in Peking Union Medical College Hospital from June 1983 to June 2002 were studied. There were 107 women(66.9%) and 53 men (33.1%), with a mean age of 38.9 years (10-73 years). 100 patients were underwent SS and 148 patients were underwent US prior to surgery, and the results were compared with operative and histological findings. RESULTS: The sensitivity of SS and US in localization of the enlarged parathyroid glands was 94.0% and 85.1% respectively, and the positive predictive value of SS and US was 100% and 89.1% respectively, the overall sensitivity was 98.9% by combination of SS and US. In solitary parathyroid adenomas group (n = 145), the sensitivity of SS and US was 93.3% and 84.7% respectively; There was no significant difference (P = 0.428) in sensitivity of SS between the parathyroid glands correctly identified and undetected in classical neck location as compared with ectopic parathyroid glands, whereas significantly (P = 0.026) influenced by the US sensitivity. CONCLUSIONS: Different sensitivity exit between SS and VS in preoperative localization in patients with pHPT undergoing parathyroidectomy. The combined use of SS and US could increase the sensitivity of localization technique. Ectopic parathyroid had no influence on the sensitivity of 99mTc-MIBI scanning, but decreased the sensitivity of ultrasonography. The size of parathyroid tumors had effects on the sensitivity of ultrasonography. Otherwise, various conditions causing SS false negative were observed. Some interfere factors should be excluded when SS negative results were encountered in clinical practice. PMID- 12905741 TI - [Cloning and characterization of a novel rat gene RSD-7 differentially expressed in testis]. AB - OBJECTIVE: To isolate and identify the differentially expressed genes in spermatogenesis for the understanding molecular mechanism of spermatogenesis. METHODS: Screening of the cDNA library, Northern blot, expression and purification in E. coli with GST expression system, immunocytochemical staining of testis sections were used. RESULTS: (1) A cDNA fragment designated as RSD-7 was isolated from rat testis cDNA library. It was 1,238 bp in length, coding a protein of 232 amino acids with the GenBank accession number AF315467. The encoding protein of RSD-7 cDNA had a Ubiquitin-like domain. (2) Northern blot indicated that RSD-7 was uniquely expressed in rat testis, and in the testis RSD 7 emerged on the 30th postnatal day and expressed until 120th postnatal day. (3) Expression and purification of RSD-7 protein in E. coli with GST expression system and were used to obtain anti-RSD-7 antibody. (4) Immunolocalization of RSD 7 in rat testis revealed that it is expressed only in Sertoli cells. CONCLUSIONS: Transcription pattern of RSD-7 and localization of RSD-7 protein in testis have been made, which established the base for the functional study of RSD-7. PMID- 12905742 TI - [Model of B immunoblastic lymphomas in the Hu-PBL-SCID mice]. AB - OBJECTIVE: To constitute a model of B immunoblastic lymphomas in the Hu-PBL-SCID mice. METHODS: The SCID mice were reconstituted by intraperitoneal injection (i.p.) of 5 x 10(7) human lymphocytes from Epstein-Barr virus (EBV) seronegative individuals. After one week, the SCID mice were inoculated with EBV by i.p. injection, and subjected to the investigation of whether there was any tumor in the abdomen of such SCID mice four weeks later. The characteristics of the found tumor was observed by the methods of Hematoxylin-eosin (HE) stain, immunohistochemical staining and polymerase chain reaction (PCR). RESULTS: Compared with the control groups, all the EBV-infected Hu-PBL-SCID mice had abdominal solid tumors [(32 +/- 12.5) mm3] developed, often located in the liver. HE staining and immunohistochemical staining showed the tumors were human B cell lymphomas. EBV DNA could be detected in the tumors by the PCR. CONCLUSIONS: The model of B immunoblastic lymphomas in the Hu-PBL-SCID mice is successfully constituted, and may well be useful to the human tumor immunological study. PMID- 12905743 TI - [Effect of G protein in the dual regulation of opioid receptor agonist on the delayed rectified potassium channels]. AB - OBJECTIVE: To observe the role of G protein in the dual regulation of opioid receptor agonist on the delayed rectified potassium channels. METHODS: Using whole-cell patch-clamp techniques applied to NG108-15 cells, investigate the effect of opioid receptor agonist on the delayed rectified potassium channels by administration of Guanosine-5'-0'-2-thiociphosphate (GDP beta S), Pertusis Toxin (PTX), Tetroacetic acid nueleoside diphosphate kinase (NDPK) and Adenosine-3' 5' cyclic monophosphate cAMP in the pipette solution. RESULTS: (1) GDP beta S could block the changes induced by both high and low concentration of (D-Pen2.5) enkephalin (DPDPE) (P < 0.05). (2) PTX could inhibit the excitative regulation on K+ channel by high concentration of DPDPE (P < 0.05). But CTX had no effect on K+ channel caused by DPDPE. (3) UDP could block the excitative effect of K+ channel by high concentration of NDPK, while have no changes on the inhibitory effect caused by low concentration of opioid agonists. (4) cAMP took part in the regulation in high concentration of agonist administration (P < 0.05), while no changes for low concentration of agonists. CONCLUSIONS: Dual changes were observed on delayed rectifier potassium channel by agonist treatment on NG108-15 cells. The excitative effect was Gi/o coupled in high concentration of agonist incubation, related to cAMP. While the inhibitory effect was possibly induced by G protein beta gamma subunit directly. PMID- 12905744 TI - [Enhancement of human papillomavirus type 16E6E7 vaccine-induced specific immune response by coimmunization with B7-1 co-stimulatory gene]. AB - OBJECTIVE: To develop a therapeutic vaccine against human tumors associated with human papillomavirus type 16E6E7 (HPV16E6E7) which is modified from a Chinese patient of the cervical cancer which possessing the antigenicity and no transforming activity, and explore more active vaccine for inducing cellular immunity with mouse co-stimulatory molecular B7-1 gene. METHODS: The modified E6E7 gene expression plasmid pVR1012-fmE6E7 was constructed and transfected Cos-7 cells, and the E7 protein specific expression was testified by immunofluorescence assay. C57BL/6 mice were immunized intramuscularly with pVR1012-fmE6E7 alone or in combination with B7-1 gene expression plasmid (pcDNA3.1-B7-1). The activity of cytotoxic T lymphocytes (CTLs) was analyzed with 51Cr specific release assay and the specific antibody in sera was analyzed by indirect ELISA. HPV16 positive C57BL/6 tumor cells C3 were inoculated subcutaneously in the vaccinated mice to assay the growth of transplanted tumors. RESULTS: The specific CTLs and antibody from immunized mice were induced efficaciously by the E6E7 gene immunization, and co-administration of B7-1 gene could significantly enhanced the CTLs immune responses of fmE6E7, and protected 33% immunized mice against C3 tumor cells challenge. In contrast, all the mice immunized only with fmE6E7 gene developed transplanted tumors after C3 cells challenge. There was no difference in E7 specific antibody responses between mice immunized with the E6E7 gene only and co administration with B7-1 gene. CONCLUSIONS: The modified E6E7 gene can be used as target gene for developing DNA vaccine, and B7-1 gene may represent an attractive adjuvant for enhancement of the specific cellular immune responses. PMID- 12905745 TI - [Effect of AP-1 on expression of plasminogen activator inhibitor-1 in human vascular endothelial cells]. AB - OBJECTIVE: To study the effect of nuclear transcription factor AP-1 on tumor necrosis factor alpha (TNF-alpha) or minimal modified low density lipoprotein (mmLDL)-induced expression of plasminogen activator inhibitor-1 (PAI-1) in human vascular endothelial cells. METHODS: Using gene recombination techniques, four luciferase reporter gene plasmids containing different length of human PAI-1 gene promoter were constructed. Through the transient transfection analysis, the roles of AP-1 element (from -823 bp to -553 bp) in PAI-1 promoter have been determined. In order to further verify the role of AP-1 element, the three site-directed mutants were recovered using PCR and sequencing assay. RESULTS: The induction by TNF-alpha or mmLDL were decreased markedly when the three AP-1 elements in PAI-1 promoter had been mutated respectively. CONCLUSIONS: These results indicate that the AP-1 element in PAI-1 promoter may have important role in PAI-1 gene transcriptions in endothelial cells induced by TNF-alpha or mmLDL. PMID- 12905746 TI - [Efficient generation of recombinant adenoviruses expressing antiangiogenic fragment of human thrombospondin1]. AB - OBJECTIVE: To construct recombinant adenoviruses expressing antiangiogenic fragment of human thrombospondin1 (TSP1f). METHODS: TSP1f cDNA was amplified by RT-PCR from normal human peripheral blood mononuclear cells and was subcloned into a shuttle vector pShuttle-CMV. After sequence confirmation, the resultant plasmid was linearized by the restriction endonuclease Pme I and cotransformed with the supercoiled adenoviral vector pAdEasy-1 into Escherichia coli strain BJ5183. Recombinants were selected by Kanamycin resistance and screened by restriction endonuclease digestion. Then, the recombinant adenoviral construct was cleaved with Pac I and transfected into the packaging cell line 293. The adenoviral vector ADV-TSP1f was propagated in 293 cells and purified by cesium chloride (CsCl) density centrifugation. PCR and Western blot analysis were performed to confirm TSP1f expression. RESULTS: Of 43 Kanamycin-resistant colonies obtained from cotransformation, all of the 10 smallest ones were the correct recombinants. TSP-1f was expressed efficiently by ADV-TSP1f. The virus stock titer after CsCl banding was 1.0 x 10(11) pfu/mL. CONCLUSIONS: Generating recombinant adenoviruses using AdEasy System results in highly efficient viral production and significantly decrease the time required to construct usable viruses. ADV-TSP1f can be further used in in vivo gene therapy studies. PMID- 12905747 TI - [Inhibition of 6A8 alpha-manosidase expression induces decrease of adhesion to laminin and reduction of lamellipodia of human nasopharyngeal carcinoma cell CNE 2L2]. AB - OBJECTIVE: To investigate the inhibitory effect of 6A8 alpha-manosidase expression on the adhesiveness of CNE-2L2 cells to laminin and the lamellipodia on cell surface. METHODS: 6A8 alpha-manosidase expression was detected by Western blotting. For assaying the adhesion of cells to laminin, cells were incubated in laminin-coated plate at 37 degrees C for 1 h, the adhered cells were stained with crystal purple dissolved in 0.1 mol/L Sodium Citrate/50% ethanol. Absorbance 540 nm was measured. Adhesion rate (R) was calculated according to formula R = AT/A100 x 100%. Here A100 represents 100% adhesion. lamellipodia on cell surface was observed upon a scanning electron microscopy. RESULTS: The adhesion rate of two clones (AS1 and AS2) with inhibition of 6A8 alpha-manosidase expression to laminin was 0.447 +/- 0.096 and 0.533 +/- 0.065 respectively. The adhesion rate of three controls with normal expression of 6A8 alpha-manosidase to laminin was 0.78 +/- 0.035, 0.7 +/- 0.05 and 0.80 +/- 0.04 respectively. The difference was significant (P < 0.01). CNE-2L2 cells with normal expression of 6A8 alpha manosidase was rich in lamellipodia on their surface. Lamellipodia nearly disappeared on the cells with inhibition of 6A8 alpha-manosidase expression. CONCLUSIONS: Inhibition of 6A8 alpha-manosidase expression results in decrease of adhesion to laminin and reduction of lamellipodia of human nasopharyngeal carcinoma cell CNE-2L2. PMID- 12905748 TI - [Acute lung injury and changes of myocardial ATP enzymes induced by lipopolysaccharide in aging rats]. AB - OBJECTIVE: To investigate whether acute lung injury (ALI) and changes of myocardial ATP enzymes were induced by intravenous or intraventricle of left heart injection of lipopolysaccharide (LPS) in aging rats. METHODS: 40 male Wistar rats were used for reproducing aging animal model. Aging rats were randomly divided into aging control group (n = 8), ALI group (LPS, 5 mg/kg body weight intravenous injection, n = 16), and LPS group (same dosage LPS, intraventricle of left heart injection, n = 16). The samples (blood, lung and heart) were collected at 2, 6 hours after LPS or saline administration. RESULTS: Compared with aging control, protein content in bronchial alveolar lavage fluid (BALF), ratio of lung wet/dry weight and the LA, NO2-/NO3- and MDA contents in blood were increased markedly (P < 0.01) at 2, 6 hours in ALI group. The GSH-Px, Na(+)-K(+)-ATPase activities in lung tissue were decreased significantly (P < 0.01), but NO2-/NO3- content in lung tissue was increased obviously (P < 0.01) at 2 hours in ALI group. These changes were maintained until at 6 hours after LPS administration. The above parameters were no obviously changes in myocardium at 2 hours after LPS administration in ALI group. But at 6 hours, MDA content was increased obviously (P < 0.01); Na(+)-K(+)-ATPase, Ca(2+)-Mg(2+)-ATPase and GSH Px activities in myocardium were decreased markedly (P < 0.01). While in LPS group, only NO2-/NO3- contents were increased (P < 0.05) in blood and lung tissue as well as Na(+)-K(+)-ATPase activity in lung tissue were decreased (P < 0.05), another parameters had no obvious changes. CONCLUSIONS: ALI was obviously formed by intravenous injection LPS after 2, 6 hours in aging rats. Myocardial enzyme etc decreased only at 6 hours in ALI group. But above parameters were no obviously changes in LPS group. It was suggested that there was probable myocardial damage in rats of ALI group, and it was mainly induced by ALI. PMID- 12905750 TI - [A randomized, double blind, and controlled clinical trial of the non-addictive propacetamol in postoperative analgesia]. AB - OBJECTIVE: To compare the postoperative analgesic efficacy and safety of the non addictive propacetamol hydrochloride (Pro-Bufferin) injection and dolantin in a prospective, randomized, double blind and controlled clinical trial. METHODS: After the pain intensity was assessed when the patients were undergone thoracic and abdominal selective surgery became fully conscious, 40 consecutive patients with moderate to severe postoperative pain (equivalent to Pain Grade I and II of American Anesthesia Association classification) were randomized into the study against the control groups. The two groups were similar for age, sex, height/weight, disease categories, operation categories, anesthesia methods and duration, vital signs, hepatorenal function, and blood cell count (P = 0.06 0.93). In the study group, 2 g propacetamol in 100 ml normal saline (NS) intravenously with 1.0 ml NS intramuscularly as the placebo control to dolantin were administered. In the control group, 1.6 g mannitose in 100 ml NS intravenously as the placebo control to propacetamol with 50 mg dolantin (1.0 ml) intramuscularly as the positive control to propacetamol were administered. The intensity change of postoperative pain was then evaluated 10 times with visual analog scale and verbal describing scale during 6 h from the beginning of propacetamol infusion. Vital signs and adverse reactions were also documented. After all data were put into the computer, the blinding codes were decoded and the statistic analysis was then made. RESULTS: There was no significant difference (P = 0.93) about the area under the curve of "Pain Relieve Score vs. Time". The "starting to effect" time (15-30 min), analgesic duration (6 h) and the percentage of excellent or good analgesic effect (90%) in the two groups were the same. Adverse reactions didn't reached the statistic different level (P = 0.35). CONCLUSIONS: Propacetamol HCL injection 2 g intravenously could be an alternative to dolantin 50 mg intramuscularly for moderate to severe postoperative pain with its advantage of being non-addictive. PMID- 12905749 TI - [Construction of a recombinant human adenovirus expressing the ORF2 antigen of HEV and immunization of mice by mucosal system]. AB - OBJECTIVE: To construct a replication-defective recombinant adenovirus expressing the ORF2 (112-660aa) antigen of hepatitis E virus (HEV) and evaluate its immunization effect in BALB/c mice by mucosal inoculation. METHODS: The HEV ORF2 gene encoding for 112-660aa was amplified from plasmid pUC-HEV and inserted into the transfer vector pTrack-CMV. The recombinant plasmid and adenoviral backbone plasmid pAdEasy-1 were co-transformed into E. coli strain BJ5183. Taking the advantage of the high efficient homologous recombination machinery presented in bacteria, the recombinant adenovirus backbone plasmid was generated in BJ5183, and then was transfected into 293 cells. Recombinant Adenoviruses were propagated in 293 cells with high titers. 8-week-old BALB/c mice were inoculated intraperitoneally and intranasally with 10(7) pfu recombinant adenovirus each on weeks 0, 3, 5, 7, 10. RESULTS: Both groups of mice induced humoral IgG immune response with the highest titers 1:1,000 and 1:10,000 each. Only the group inoculated intranasally could induce mucosal IgA immune response. CONCLUSIONS: The adenoviral recombinant can stimulate specific humoral and mucosal immune response in mice and is potentially to be used as a candidate vaccine for the treatment of HEV infection. PMID- 12905751 TI - [Effect of vitamin E on memory and brain monoaminergic neurotransmitter in chronic episodic hypoxia rat]. AB - OBJECTIVE: To investigate the effect of Vitamin E (VitE) on memory and brain monoaminergic neurotransmitter level in chronic episodic hypoxia (EHYP) rat. METHODS: VitE [50 IU/ (250 g.d) or 5 IU/ (250 g.d)] was given to the EHYP rat model. The memory was evaluated by the passive avoidance test and the levels of monoaminergic neurotransmitter, including norepinephrine (NE), dopamine (DA) and 5-hydroxytryptamine (5-HT), were determined in three different brain regions (including cerebral cortex, hippocampus and striatum) using high performance liquid chromatography and electrochemical detection (HPLC-ECD). RESULTS: The performance on passive avoidance test of EHYP rats was worse than that of controlled rats (P < 0.01). The performance of rats in two different treatment groups was better than that of EHYP rats (P < 0.05), the performance of rats in high-dose group was worse than that of rats in low-dose group (P < 0.05). Compared with controlled rats, levels of monoaminergic neurotransmitters in different brain regions of EHYP rats decreased significantly (P < 0.05). Compared with EHYP rats, level of NE and DA in cerebral cortex and level of monoamine (NE, DA, and 5-HT) in hippocampus and striatum of low-dose treated rats were increased significantly (P < 0.05). Different with low-dose treated rats, only level of monoamine (NE, DA, and 5-HT) in striatum and level of 5-HT in hippocampus in high dose treated rats were increased significantly (P < 0.05), as compared with the EHYP rats. CONCLUSIONS: vitE can improve memory and increase brain monoaminergic neurotransmitter of EHYP rats. Moreover, the effect of low-dose vitE is better than that of high-dose VitE. PMID- 12905752 TI - [Comparison of SIB-IMRT treatment plans for upper esophageal carcinoma]. AB - OBJECTIVE: To implement simultaneous integrated boost intensity-modulated radiotherapy(SIB-IMRT) plans for upper esophageal carcinoma and investigate the dose profiles of tumor and electively treated region and the dose to organs at risk (OARs). METHODS: SIB-IMRT plans were designed for two patients with upper esophageal carcinoma. Two target volumes were predefined: PTV1, the target volume of the primary lesion, which was given to 67.2 Gy, and PTV2, the target volume of electively treated region, which was given to 50.4 Gy. With the same dose-volume constraints, but different beams arrangements (3, 5, 7, or 9 equispaced coplanar beams), four plans were generated. Indices, including dose distribution, dose volume histogram (DVH) and conformity index, were used for comparison of these plans. RESULTS: The plan with three intensity-modulated beams could produce good dose distribution for the two target volumes. The dose conformity to targets and the dose to OARs were improved as the beam number increased. The dose distributions in targets changed little when the beam number increased from 7 to 9. CONCLUSIONS: Five to seven intensity-modulated beams can produce desirable dose distributions for simultaneous integrated boost (SIB) treatment for upper esophageal carcinoma. The primary tumor can get higher equivalent dose by SIB treatments. It is easier and more efficient to design plans with equispaced coplanar beams. The efficacy of SIB-IMRT remains to be determined by the clinical outcome. PMID- 12905753 TI - [Action of miosis of 1% pilocarpine liposome in rabbits]. AB - OBJECTIVE: To compare the action of miosis of 1% pilocarpine liposome with 1% pilocarpine solution in rabbits. METHODS: 18 white rabbits were randomly divided into 3 groups. Test group received 1% pilocarpine liposome, positive control group received 1% pilocarpine solution, negative control group received liposome. Each eye drop instilled into left eye of rabbits and sterile saline solution instilled into right eye as control. The pupil diameter was measured at time intervals of beginning, 0.25, 0.5, 1, 2, 3, 4, 5, 7 hours. RESULTS: The mean pupil diameter change of 3 groups in both eyes was not significant (P > 0.05) at beginning. The strongest action of miosis took place 0.25 h in positive control group and 0.5 h in test group after instillation. The dilation of pupil in both groups took place 1 h and 3 h, and the restoration of pupil in both groups took place at 5 h and 7 h. The mean pupil diameter of negative control group was not significant in seven hours. CONCLUSIONS: The results suggest that 1% pilocarpine liposome improves the bioavailability and prolong the duration of its action. PMID- 12905754 TI - [Vitamin K and osteoporosis]. AB - There is a closely relationship between vitamin K and osteoporosis. As a cofactor for carboxylase activity, vitamin K can facilitate the conversion of glutamyl to gamma-carboxyglutamyl residues and influence the synthesis and excretion of gamma carboxylation of osteocalcin to increase the formation of bone. Vitamin K can also effectively inhibit the absorption of bone mass. Besides, there are increasing evidences that vitamin K can effect the synthesis and excretion of nephrocalcin and interlukin-1,6 that can regulate calcium balance and bone metabolism. Meanwhile, there is a consistent line of evidence in human epidemiologic and intervention studies that clearly demonstrate that vitamin K can not only increase bone mineral density in osteoporotic people, but also reduce fracture rates to improve bony health. However more researches are required before vitamin K is widely applied in prevention and treatment of osteoporosis. The American Medical Association recently has increased the dietary reference intakes of vitamin K to 90 mg/d for females and 120 mg/d for males. PMID- 12905755 TI - [Diagnosis and treatment of sellar germinoma]. AB - That eleven patients with sellar germinomas were confirmed by pathological examinations were retrospectively reviewed. In the 11 patients, 2 were males and 9 were females. Mean age was 16.8 years old and mean history was 2.6 years. Among them, 11 patients had diabetes insipidus, 6 patients had visual disturbances, 5 patients had retarded growth, and 5 patients had hypopituitary function. The diameter of tumor was 1-3 cm in 8 cases and greater than 3 cm in 3 cases. MRI showed hypointensity or homogeneous on T1-weighted images in 8 cases. The tumors were fibrous, purple grey or reddish in color with rich blood supply. Germinomas were confirmed by postoperative pathological examinations. All patients received postoperative irradiation. Postoperative follow-up was from 3 months to 6 years. Diabetes insipidus was improved in 9 cases. Visual function were improved in 4 cases, but hypopituitarism and retarded growth were unchanged. In conclusion, when MRI shows sellar parenchyma tumor with unclear boundary in young patients with diabetes insipidus and hypopituitarism, sellar germinoma should be highly suspected. Otherwise, radiation therapy may profitably aid to confirm the diagnosis since irradiation is the main treatment of sellar germinomas. PMID- 12905756 TI - [Hyponatremia after surgery of pituitary adenomas]. AB - OBJECTIVE: To clarify the frequency and presentation associated factors, pathogenetic mechanism, treatment and outcome of hyponatremia after surgery of pituitary adenoma. METHODS: Retrospectively reviewed the data of 186 patients who underwent pituitary surgery between January 1999 and June 2000 in the department. RESULTS: 72 (38.7%, 72/186) patients had suffered post-operative hyponatremia. Among them, 43 (59.7%, 43/72) cases the hyponatremia appeared at 4 to 7 days postoperatively. 43 (59.7%, 43/72) cases presented with nausea, vomiting, headache, dizziness, confusion and weakness. The symptoms of hyponatremia was related to the age, tumor size and adenoma cell type, but not to the sex, surgical approach and degree of removal. Treatment consisted of salt replacement and mild fluid restriction in 4 patients and salt and fluid replacement in 68 patients. Hyponatremia had been resolved within 7 days in 63 patients. CONCLUSIONS: Hyponatremia often appeared about 7 days after surgery of pituitary adenoma, especially in elderly and in patients with macroadenomas. The principle of treatment was salt and fluid replacement. PMID- 12905757 TI - [SARS epidemic trends in Beijing since late April 2003]. AB - Since May 8th, Beijing's severe acute respiratory syndrome (SARS) epidemic is on a continuous decline. Indicating and verifying measures taken since the end of April to be effective. Three stages of SARS have been distinguished in Beijing since late April, they are described in this article as rapid growth stage beginning late April until early May, decline and stable stage throughout the first week of May, and rapid decline stage since May 9th until present. These three stages depict the effectiveness of interventive measures against SARS in Beijing since late April. Information transparency towards the public is an import part among all measures taken, enhancing public awareness and understanding of the current situation and government action as well as introduce public action. Public awareness is an important factor, forming active support for government interventions. PMID- 12905758 TI - [Lung pathology of severe acute respiratory syndrome]. AB - OBJECTIVE: To study the morphological features of the lungs obtained from autopsies of severe acute respiratory syndrome (SARS) patients. METHODS: Bilateral lungs from 7 patients died from SARS were carefully studied grossly and microscopically. All tissues from these cases were routinely processed and carefully studied. RESULTS: All lungs from these cases were extremely expanded and became solid. Microscopically, the edema and fibrin exudates in the alveoli was the most common findings, especially in the early phase of the disease. The hyaline membrane was almost always present in the lungs of these cases. The organization of intra-alveolar fibrin exudates along with the interstitial fibrosis led to obliteration of alveoli and consolidation of lungs. The desquamation and hyperplasia of alveolar lining cells was also apparent. Foci of haemorrhage and lobular pneumonia, even diffuse fungal infection were frequently seen in these specimens. Micro-thrombus were easily found in these lungs. CONCLUSIONS: The lung of SARS from autopsy is characterized by edema, intra alveolar fibrin exudates, hyaline membrane formation, organization of intra alveolar exudates and fibrosis, which lead to the obliteration of alveoli and consolidation of lungs. PMID- 12905759 TI - [Clinical nutrition support and relationship of blood glucose level/insulin administration with outcome in critical SARS patients]. AB - OBJECTIVE: To evaluate the use of clinical nutritional support in critical SARS patients, and the relationship between blood glucose levels/insulin administration amount and outcome. METHODS: Twenty-one SARS patients who reached the standard of Ministry of Health's "critical level" were transferred into our ICU in an average of 11 days after onset and enrolled in this clinical trial. All patients underwent respiratory support and clinical nutrition support as scheduled. For about 60 kg patient per day 3347.2 kJ(800 kcal), 36 g protein, and 125 g carbohydrate was given intravenously; 4184 kJ(1000 kcal), 38 g protein, and 125 g carbohydrate was provided by enteral route. MCT/LCT as fat resource shared 50% calories intake. All patients received similar doses of intravenous Methylprednisolone(about 200 mg/d). Blood glucose, serum albumin, blood lymphocyte counts, and serum alanine transminase (ALT) were checked on the first admission day in ICU and on the 12th day after nutrition therapy was started. Insulin was started to pump in to maintain the blood glucose levels between 4.44 7.78 mmol/L (80-140 mg/dl) when the levels exceeded normal range. RESULTS: Upon admission into ICU, all patients had poor nutrients intake for an average of 11 days and 16 patients (76.2%) were diagnosed as malnutrition. Parenteral and enteral nutrition therapy were then offered for an average of 12 days. On the 12th day, the serum albumin increased [(28.5 +/- 2.2)] g/L vs (37.0 +/- 4.1) g/L] (P = 0.0001) and so did the lymphocytes count [(0.74 +/- 0.47)] x 10(9)/L vs (1.22 +/- 0.73) x 10(9)/L] (P = 0.02). The blood glucose maintained at lower level in the surviving patients when compared with those who died [(9.5 +/- 2.3) mmol/L vs (6.3 +/- 1.8) mmol/L] [(196 +/- 70) mg/dl vs (110 +/- 21) mg/dl] (P = 0.0002), and the abnormally high ALT levels presented in some of the patients decreased but not significantly (81.0% vs 57.1%) (P = 0.18). In order to keep blood glucose within the range 4.44-7.78 mmol/L (80-140 mg/dl), only 18.8% of the surviving patients needed insulin intervention as opposed to 80.0% of those who died (P = 0.03). The amount of insulin used in the surviving group was significant lower than that in the group who died [(24 +/- 2) IU/d vs (72 +/- 9) IU/d] (P = 0.01). CONCLUSIONS: Eleven days after SARS onset, most of the critical patients presented with malnutrition. Some improved nutrition related parameters may be associated with clinical nutritional support. The surviving patients required less insulin when compared to those who died. 80.0% of the patients who died need insulin versus only 18.8% of the surviving patients. Due to the difficulty of SARS management, this study was not a randomized controlled clinical trial. More clinical trials will be needed for checking the results of this investigation. PMID- 12905760 TI - [Detection of SARS-CoV RNA in stool samples of SARS patients by nest RT-PCR and its clinical value]. AB - OBJECTIVE: To investigate of severe acute respiratory syndrome (SARS) convalescent stool shedding by RT-PCR. METHODS: One hundred and three stool samples from 46 SARS patients were collected on May 16th, 20th, and 23rd, 2003. For each sample, RNA was extracted using commercial kit and 7 Nest RT-PCR using a 14-pair different SARS-associated coronavirus (SARS-CoV) special primers were carried out simultaneously. RESULTS: Among these 46 SARS patients, 17 cases (37.0%) were stool SARS-CoV RT-PCR negative, and 29 cases (63.0%) were SARS-CoV RT-PCR positive. The duration of positive cases lasted (31.76 +/- 10.78) d (12-64 d). The longest stool shedding case in this study lasted 64 days. Two serial stool samples and for each sample 2 RT-PCR tests using different primers were positive in this case. CONCLUSIONS: Our study observed longest stool shedding of SARS patients to be 64 days after initial onset of SARS. The average stool shedding was 32 days. Hence it is important to think highly of SARS convalescent patient stool sterilization. PMID- 12905761 TI - [Right attitude towards the postmenopausal hormone replacement therapy]. AB - We have gone through decades using hormone replacement therapy (HRT). The first problem encountered was increased endometrial cancer and solved by addition of progesterone. Now we are facing cardiovascular complications and how could we solve in the use of HRT. Research in vitro with HUAR and HUVEC and clinically seemed to show that small physiological doses might be the solution in protection of CVD. PMID- 12905762 TI - [A nationwide retrospective analysis on chronic diabetic complications and related macrovascular diseases of in-patients with diabetes during 1991-2000]. AB - OBJECTIVE: To investigate the prevalence rates of chronic diabetic complications and their risk factors. METHODS: We analyzed retrospectively the clinical and laboratory data on the 24,496 in-patients from 30 provinces, municipalities, and autonomous regions during January 1, 1991 to December 31, 2000. RESULTS: The prevalence rates of chronic diabetic complications and related macrovascular diseases were 31.9% for hypertension, 12.2% for cerebrovascular diseases, 15.9% for cardiovascular diseases, 5.0% for lower extremity vascular diseases, 34.3% for retinopathy, 33.6% for nephropathy, and 60.3% for neuropathy, total rate of chronic diabetic complications and macrovascular diseases was 73.2%. The risk factors related to all kinds of chronic diabetic complications and macrovascular diseases were age, illness duration, and systolic blood pressure. CONCLUSIONS: The prevalence rates of chronic diabetic complications of in-patients with diabetes mellitus are rather high, near to their level in developed countries. The controllable risk factor of chronic diabetic complications is systolic blood pressure. PMID- 12905763 TI - [Epidemiological survey of chronic vascular complications of type 2 diabetic in patients in four municipalities]. AB - OBJECTIVE: To provide an epidemiological description and risk factors of chronic vascular complications of type 2 diabetic in-patients in four municipalities including Beijing, Shanghai, Tianjin, and Chongqing. METHODS: Data of the study came from 3,469 Type 2 diabetic in-patients from 1991 to 2000 in 10 medical centers of Beijing, Shanghai, Tianjin, and Chongqing. A variety of parameters of in-patients were evaluated retrospectively to know the prevalence and risk factors of chronic vascular complications in the study patients. RESULTS: Overall, the detailed prevalence of chronic vascular complications is listed as follows: diabetic retinopathy 31.5%, diabetic nephropathy 39.7%, diabetic neuropathy 51.1%, hypertension 41.8%, coronary heart disease (CHD) 25.1%, cerebral vascular disease (CVD) 17.3%, vessel complication of lower limbs 9.3%. Multivariate logistic regression analysis shows that diabetes family history, duration of diabetes (> 5 years), and systolic blood pressure (> 125 mmHg) are the risk factors for diabetic retinopathy; duration of diabetes (> 5 years), systolic blood pressure (> 125 mmHg), LDL-C (> 3.12 mmol/L), and triglyceride (> 1.70 mmol/L) are the risk factors for diabetic nephropathy; age (> 45 years), duration of diabetes (> 5 years), HbA1C (> 7.0%), systolic blood pressure (> 125 mmHg), and LDL-C (> 3.12 mmol/L), are the risk factors for CHD; age (> 45 years), duration of diabetes (> 5 years), systolic blood pressure (> 125 mmHg), and triglyceride (> 1.70 mmol/L) are the risk factors for CVD. CONCLUSION: In order to improve patients' outcome, multiple metabolic controls in type 2 diabetic patients are urgently needed, which include decreasing glycemia, reducing hypertension and improving lipid levels. PMID- 12905764 TI - [Lipids disorder, macrovascular diseases, and insulin resistance in Chinese diabetic patients]. AB - OBJECTIVE: To investigate the types of lipid disorder and its relationship with macrovascular diseases and insulin resistance in Chinese diabetic patients. METHODS: 2,430 diabetic patients finished the diabetes complications assessment and were surveyed on their blood lipids, insulin level and macrovascular diseases. The insulin resistance (HOMA-IR) was calculated based on HOMA model. RESULTS: Prevalence of lipids disorder was as high as 63.8% in this group, including 23.9% with both hypercholesterolemia and hypertriglyceridemia (HY-C-T), 16.1% with hypercholesterolemia only (HY-C), 15.0% with hypertriglyceridemia only (HY-T), 5.5% with lower HDL level (L-HDL), 3.3% with hypercholesterolemia, hypertriglyceridemia and lower HDL level together. Compared with the patients with normal lipid level (control group), no duration differences existed among these groups. There were more female patients in HY-C and HY-C-T groups. BMI and WHR were higher in all groups with lipid disorder except in HY-C group. Macrovascular diseases were more common in HY-C-T patients than in control group (33.6% vs 24.0%, P < 0.001). Average blood pressure and the prevalence of hypertension were significantly higher in HY-C-T, HY-C, HY-T groups than in control group (139/79, 138/76, 134/77 vs. 132/75 mmHg; 53.2%, 50.1%, 46.2% vs. 39.2%). Fasting insulin level was significantly higher in HY-C-T and HY-C-T-L-LDL groups than in controls. Insulin resistance was more severe in all patients with lipid disorder except in L-HDL group, particularly in the HY-C-T-L-HDL group. CONCLUSIONS: (1) BMI, WHR increased significantly with the severity of lipid disorder. (2) Nearly two thirds of Chinese diabetic patients have lipid disorder, which is the strong risk factor of macrovascular diseases and aggravates insulin resistance. (3) There was severe insulin resistance in patients with hypertriglyceridemia or with hypertriglyceridemia plus other lipid disorder. PMID- 12905765 TI - [Study of the gene expression pattern in the Kkay mouse with type 2 diabetes by genomic-scale gene expression analysis]. AB - OBJECTIVE: To study the genes differentially expressed in the liver of Kkay diabetic and normal mice by genomic-scale gene expression analysis. METHODS: cDNA microarray chips containing 8,192 cDNAs were used to explore the gene expression pattern of Kkay mouse liver. RESULTS: One hundred and fifty-four genes were screened out, including 68 complete cDNAs and expressed sequence tags, and among them 40 genes were up-regulated and 114 genes were down-regulated respectively. CONCLUSION: Most of the gene expression analysis results were consistent with previous study, and the gene expression pattern of Kkay mouse based on cDNA microarray could be used for high-throughout screening out the genes associated with type 2 diabetes. PMID- 12905766 TI - [A novel gene involved in blood glucose regulation]. AB - OBJECTIVE: To clonea a novel gene related to blood glucose regulation. METHODS: Contrast rat model of autonomous regulation through jugular vein right atrium intubation of high concentration of glucose, the control rats were injected with 0.9% NaCl and skeletal muscle was separated. The differentially expressed fragments were identified by differential display technology (DDRT-PCR). After slot blot and Northern blot analysis, we excluded the artificial positive fragments and got the true EST (expression sequence tag) differentially expressed. These positive EST were used as probes to screen cDNA library of rat skeletal muscle, the coding region of full-length gene was cloned to pEGFP and L 6TG cell line was cultured and transiently transfected with the lipofectamine reagent. After 48 h, intact cells were examined with fluorescence microscope. RESULTS: We got a novel full-length cDNA, named as Fang-1. GenBank Accession No. is AF399873. Using blast software (NCBI), we found Fang-1 is rat homologue of human AK001644. They share 82% identical nucleotides, indicating the family proteins are very conserved. After high concentration of glucose stimulation compared with control rats, the expression of Fang-1 was up-regulated and the expression product of Fang-1 was localized in both cytoplasm and cell nucleus. CONCLUSIONS: A novel gene related to blood glucose regulation was cloned from rat skeletal muscle. The expression product of Fang-1 was localized in both cytoplasm and cell nucleus. The gene can regulate blood glucose level by controlling some mechanisms unknown now with down-regulation expression. PMID- 12905767 TI - [Functional analysis of the single nucleotide polymorphisms in the PRKCZ gene]. AB - OBJECTIVE: To study the function of 5 single nucleotide polymorphisms (SNPs) of the PRKCZ gene, a susceptibility gene for type 2 diabetes in Han population of North China, in the pathogenesis of the disease. METHODS: Bioinformatic methods and reporter gene activity determination were used to analyze the function of the 5 SNPs. RESULTS: The reporter gene activities of different alleles of 2 SNPs, rs427811 and rs809912, were obviously different, which implies that these 2 SNPs might be susceptibility loci of the disease. CONCLUSION: The PRKCZ gene is further confirmed to be a susceptibility gene for type 2 diabetes in Han population of North China. Two SNPs in the gene play a role in the pathogenesis of the disease by affecting the expression level of PRKCZ gene. PMID- 12905768 TI - [Linkage disequilibrium analysis of the single nucleotide polymorphisms in the PRKCZ gene]. AB - OBJECTIVE: To search for the disease-associated haplotype in the PRKCZ gene, a susceptibility gene for type 2 diabetes in Han population of North China, by case control study and linkage disequilibrium (LD) analysis using single nucleotide polymorphisms (SNPs). METHODS: SNPs located in the PRKCZ gene were chosen from public SNP domain by bioinformatic methods and single base extension (SBE) method was used to genotype the loci in 173 sporadic type 2 diabetes patients and 152 normal individuals to perform case-control study and LD analysis. Haplotype block were constructed in these populations. RESULTS: Several SNPs in the PRKCZ gene were found to be associated with the disease. The SNPs formed different haplotype block pattern in case and control groups. The frequencies of the haplotypes formed by 5 SNPs were statistically different between the two groups. CONCLUSION: The haplotype formed by 5 SNPs in the PRKCZ gene may be associated with type 2 diabetes in Han population of China, which is confirmed from statistics to be a susceptibility gene for the disease. PMID- 12905769 TI - [Compensated hyperinsulinemia based on selective insulin resistance predicts elevation of blood pressure in non-diabetic adults]. AB - OBJECTIVES: To investigate if hyperinsulinemia or insulin resistance could predict the elevation of blood pressure in non-diabetic adults. METHODS: One hundred and seventy non-diabetic adults (NGT 107, IGT 63) were included based on the screen by OGTT in 1986. Height, weight, blood pressure were measured. Plasma glucose and insulin concentration at 0.60 and 120 min during OGTT were determined at baseline. All the subjects were followed for six years with blood pressure and plasma glucose examined at the end of the study. Subjects worsening to diabetes were excluded. Insulin area under-curve (INSAUC) and insulin sensitivity index [IAI = (1/FINS x FPG)] were calculated. Stepwise regression analysis was performed to evaluate the effects of INSAUC and insulin sensitivity to the elevation of blood pressure. RESULTS: Both SBP and DBP levels at the end of the study were increased with increased INSAUC baseline. The SBP were (119.5 +/- 2.3), (122.1 +/- 2.5), (129.4 +/- 2.4) and (128.3 +/- 2.6) mmHg, and the DBP were (78.6 +/- 1.6), (79.7 +/- 1.7), (85.2 +/- 1.4) and (84.0 +/- 1.0) mmHg from the lowest to the highest quartiles of INSAUC respectively. Pearson correlation analysis showed Age, SBP, DBP, BMI, FINS, INS1h, INSAUC at baseline were positively correlated to blood pressure levels at the end of the study. After the adjustment of Age, sex, BMI, smoking, PG2 h and blood pressure at baseline, the INSAUC was significantly correlated to blood pressure six years later, while the insulin sensitivity index was not. CONCLUSION: The compensated hyperinsulinemia based on selective insulin resistance rather than insulin resistance to glucose per se could predict the elevation of blood pressure in nondiabetic adults. PMID- 12905770 TI - [Role of Fas, FasL in NOD insulitis]. AB - OBJECTIVE: To analyze the significance of Fas-FasL in NOD insulitis and to explore the mechanism of the autoimmune diabetes. METHODS: Thirty-two female NOD mice, 3-32 weeks of age, were selected. The blood glucose concentrations were recorded. The pathological data were obtained from the HE staining of the pancreatic sections and the immunohistochemical staining, in which insulin, Fas, FasL, CD8 were detected. RESULTS: Diabetes was found from the age of 14 weeks. In normal islets, insulin + cells accounted for (59.37 +/- 1.21)%, and some islet cells were observed expressing Fas. At the age of 6 weeks, insulitis lesions could be found. The average score of insulitis tended to rise with the increasing age (P < 0.0005). Meanwhile, insulin + cells decreased (P < 0.0005), and correlated negatively with scoring (P < 0.05). Fas+ islet cells increased (P < 0.0005), correlated positively with scoring (P < 0.01). In insulitis lesions, islet cells expressed FasL that increased gradually (P < 0.0005) and correlated positively with scoring (P < 0.01). The infiltrating cells were all Fas negative. But these mononucleated cells showed the expression of FasL and CD8, both increasing gradually (P < 0.0005). Furthermore, there was certain correlation between the expression of some antigens: in islet cells, between Fas and insulin (negative, P < 0.01), insulin and FasL (negative, P < 0.01), and Fas and FasL (positive, P < 0.01). In the infiltrating cells, the expression of CD8 was correlated with FasL (positively, P < 0.01); it was also found that there was a negative correlation between Fas+ islet cells and CD8+ mononucleated cells (P < 0.05). CONCLUSION: To sum up, there may be some important and complicated effects by Fas-FasL on the damage of beta cells and the regulation of autoreactive T cells in NOD insulitis, which will facilitate further studies in human type 1 diabetes. PMID- 12905771 TI - [Expression of Fas, FasL, and NF-kappa B in the process of osteoclast-like cell apoptosis effected by sodium fluoride]. AB - OBJECTIVE: To detect the changes in the expression of apoptosis signals: Fas, FasL and NF-kappa B in the process of osteoclast-like cell (OLC) apoptosis effected by sodium fluoride. METHODS: After co-culture of osteoclast-like cells with 0, 5, 10 and 15 mg/L sodium fluoride, Fas, FasL and NF-kappa B antibody expressions were detected by immune-histochemistry. RESULTS: The expression of Fas and FasL increased with the concentration of the sodium fluoride, however the expression of NF-kappa B decreased with the concentration of sodium fluoride. CONCLUSION: In the process of OLC apoptosis induced by sodium fluoride, the expression of Fas and FasL increased, and that of NF-kappa B decreased with the concentration of sodium fluoride respectively, and the changes of the expression present a dose-dependent pattern. PMID- 12905772 TI - [Comparison of capabilities of survival, proliferation and expansion between CD34+CD59+ cells from patients with PNH and CD34+ cells from normal control]. AB - OBJECTIVE: To explore in vitro expansion of CD34+CD59+ cells from patients with PNH, and compare the capabilities of survival, proliferation and expansion between CD34+CD59+ cells from patients with PNH and CD34+ cells from normal control. METHODS: CD34+CD59+ cells from patients with PNH and CD34+ cells from normal control were selected from the bone marrow mononuclear cells by means of two-step sorting method with immunomagnetic microbead-flow cytometry, then underwent in vitro expansion for two weeks and semi-solid culture in vitro before and after expansion. RESULTS: (1) CD34+CD59+ cells from patients with PNH can be expanded effectively in vitro, and the biggest expansion of CD34+CD59+ cells was about 23.49 fold on the 7th day. (2) There were some similar characteristics between CD34+CD59+ cells from patients with PNH and CD34+ cells from normal control, such as: the best combination of hematopoietic factors for in vitro expansion was SCF+ IL-3 + IL-6 + FL + Tpo + Epo, and the seventh day was the most suitable in course of 4-14 days for in vitro expansion, and after in vitro expansion, the cells remained CD59 positive and strong capability of performing colony-forming. (3) CD34+ cells from normal control had better proliferation, expansion and stronger potential to survive than CD34+CD59+ cells from patients with PNH. CONCLUSIONS: (1) In vitro expansion of CD34+CD59+ cells from patients with PNH can be performed. The present study showed the possibility of performing ABMT or APBSCT clinically for patients with PNH. (2) There were some similar characteristics between CD34+CD59+ cells from patients with PNH and CD34+ cells from normal control, but the latter had better proliferation, expansion and stronger potential to survive than the former. CD34+CD59+ cells from patients with PNH were not completely normal cells. PMID- 12905773 TI - [Endolymphatic hydrops of both ears of guinea pig produced by aldosterone]. AB - OBJECTIVE: To develop endolymphatic hydrops in both ears of guinea pigs by aldosterone. METHODS: Thirty animals were divided into 3 groups with 10 in each group. Group A received left ears operation following Kimura', with right ears non-operated as control; group B received introperitonal injection of aldosterone (0.1 mg.0.1 kg-1.d-1) for continuous 5 days; group C received introperitonal injection of 0.5 ml 0.9% NS. Five out of group A,B and C were killed 1 month and 2 months later respectively for analysis of blood K+, Ca2+, Na+ and pathological examination of inner ear, heart, lung, brain, liver and kidney. RESULTS: In group A' moderate to severe endolymphatic hydrops developed in the operated ears during 1 and 2 months. In group B, mild endolymphatic hydrops developed in the both ears during 1 month, and within 2 months it was moderate and severe. No endolymphatic hydrops developed in group C. In contrast to group C, No significant changes could be found in blood analysis of group A, but the concentrations of K+ and Ca2+ were declined, and that of Na+ were increased in group B. The heart, lung, brain, liver, and kidney of all animals were normal. CONCLUSIONS: Aldosterone may induce endolymphatic hydrops in both ears. PMID- 12905774 TI - [Comparison of radiographs and magnetic resonance imaging in the detection of carpal lesions in patients with rheumatoid arthritis]. AB - OBJECTIVE: To compare MR imaging with radiographs in detection of the erosions of the wrists in patients with rheumatoid arthritis (RA). METHODS: In 50 wrists of 25 patients with RA, radiographs of AP projection and MR imaging examination were performed within 1 week. The MRI sequences were SE T1-weighted image, FSE T2 weighted image, and GE T2* weighted image in coronal plane. Gd-DTPA enhanced SE T1-weighted image with same parameters of pre-contrast administration SE T1WI was performed in 25 wrists, which showed prominent lesions in each 25 patients. RESULTS: In 50 wrists of 25 patients with RA, bone erosions were found by MR images compared to radiographs in 46 versus 14 wrists, respectively. There were 170 sites of carpal erosions found by MR images, while radiographs found only 23 sites of carpal erosions. Among them triquetrum and lunate were the most frequently involved sites. Significant difference (P < 0.001) was statistically found between conventional radiography and MR imaging in detection of erosions of the wrists of RA. MR imaging also permitted demonstrating synovitis and joint effusion in the wrists. Gd-DTPA-enhanced images allowed distinguishing pannus changes from joint effusion. CONCLUSION: MRI is superior to conventional radiography with respect to detection of bone erosion in wrists. For those whose conventional radiographs show suspected RA, MRI examination should be advised for the early diagnosis. PMID- 12905775 TI - [Clinical analysis of 7 patients with Gerstmann syndrome secondary to cerebral vascular disease]. AB - OBJECTIVE: To analyze clinical features of patients with Gerstmann syndrome (GS). METHODS: We retrospectively analysed the clinical manifestations of 7 patients (6 men and 1 woman) with GS secondary to cerebral vascular diseases and reviewed the literatures. RESULTS: The age ranged from 51 to 70 years with a mean of 70 years. They all had sudden onset and the tetrad of GS-finger agnosia, left-right disorientation, agraphia and acalculia, 3 patients accompanied by incomplete aphasia, 3 by anomic aphasia, 2 by alexia and 1 by constructional apraxia. Cranial computed tomographic scan showed low-density focus of the left parietal lobe in 6 cases and high-density focus of the left parietal lobe in 1 case. CONCLUSION: GS has the high value in localization and the lesion is mainly localized to angular gyrus of the dominant hemisphere. PMID- 12905777 TI - [Ultrasound contrast agents in liver imaging]. AB - Ultrasound contrast agents may modify any of the three basic physical interactions between ultrasound and body tissues: absorption, reflection and refraction, and then the amount of backscatter can be increased. New imaging techniques including second harmonic imaging, interval-delay harmonic imaging, pulse-inversion harmonic imaging and sub-harmonic imaging proved to increase the sensitivity of altrasound contrast agents. There are several types of agents that have been used for liver imaging: free gas bubbles, saccharide microparticle suspensions, encapsulated microbubbles, perfluorocarbons, organ or tumor-specific agents. Ultrasound contrast agents can show characteristic vascular features of liver, specific types of hepatic tumors, or characteristic dynamic phases of perfusion imaging; therefore they can provide meaningful information for clinical diagnosis. PMID- 12905776 TI - [Nature history and genetics of type 2 diabetes]. AB - The nature history of type 2 diabetes is characterized by the interaction between beta-cells function and insulin sensitivity. Normal glucose tolerance could be maintained when there is a balance between insulin sensitivity and beta-cells function. For most of type 2 diabetic patients, both defects in insulin secretion as well as insulin action are needed for the development of diabetes. In addition, there is abundant evidence indicating the importance of genetic factors in the development of type 2 diabetes, including data from the twin studies, family studies, studies among different ethnic groups living in the same environment and admixture studies. In the last two decades, that genetic defects affecting both beta-cells function and insulin sensitivity has been identified. But those defects can only explain a very small portion of etiology of type 2 diabetes, and the gene defects contributing to the cause of most type 2 diabetes are still at large. We have learned, from the past experience that type 2 diabetes is a very heterogeneous and complex disease. The complexity of type 2 diabetes is related to factors such as genetic heterogeneity, interactions between genes, and the modulating role played by the environment. PMID- 12905778 TI - [Recombinant virus assay: a rapid assay for assessment of drug susceptibility of human immunodeficiency virus type 1 isolates]. AB - OBJECTIVE: To establish a rapid assay for assessment of drug susceptibility of human immunodeficiency virus type 1 isolates (Recombinant virus assay). METHODS: This procedure allows the generation of viable virus with SI phenotype by homologous recombination of a RT-PCR-derived pool of reverse transcriptase (RT) coding sequences into an RT-deleted, noninfectious proviral clone, pHIV delta RTBstE II. Then the drug susceptibility of recombinant virus to RT inhibitors can be assessed in the Hela CD4+ plaque reduction assays. RESULTS: Analysis of 7 HIV strains with SI or NSI phenotype showed that recombinant viruses accurately exhibited the same genotype as that of the original HIV1 isolates. The results of drug susceptibilities of HIV1 isolate got by recombinant virus assay were the same as that by standardized peripheral blood mononuclear cell culture assay. CONCLUSION: Recombinant virus assay is a rapid and accurate method to assess the drug sensitivity of HIV1 isolates with SI or NSI phenotype. PMID- 12905779 TI - [Embolization of intracranial aneurysms with new mechanically detachable coils]. AB - OBJECTIVE: To primarily embolize intracranial aneurysms with new-developed mechanically detachable coils, which is called Detachable Coil System (DCS). METHODS: Five intracranial aneurysms were embolized with DCS, in which 2 were giant aneurysms. DCS with J coils were initially used to make baskets in giant aneurysms, and then other kinds of coils were utilized to pact the aneurysmal cavity. Whereas, spiral coils were mostly used in smaller aneurysms. Coils of DCS were detached by rotating the delivering wire after the coils were satisfactorily positioned in aneurysms. RESULTS: Two of the 5 cases were totally occluded with DCS. Two was sub-totally embolized. Pushing, withdrawal, and adjusting of DCS were safe and efficient. Detachment of DCS was fast. In one case of wide-necked giant aneurysm, coils could not stay inside the aneurysm, which was treated by occlusion of the parent artery. CONCLUSIONS: DCS is a new addition to the interventional radiologist's armamentarium. Detachment is faster. J coils are suitable for giant aneurysms for its unique way of coiling. But more evaluation depends on accumulation of the clinical usage. PMID- 12905780 TI - [Positive and negative selection of thymocytes]. AB - Maturation of T cells in thymus plays key roles in antigen-specific immune response. In maturation, thymocytes undergo positive and negative selection. The MHC restriction of T cell is determined in positive selection. The tolerance of T cells to autoantigen is acquired during negative selection. Quantitation (avidity) theory points out that difference of avidity between thymocytes and stroma cells leads to difference in the signals integrated inside cells. When the signal reaches a particular threshold, positive or negative selection takes place. In this article, the selection mechanism and the relationship between positive and negative selection are mini-reviewed. PMID- 12905781 TI - [TCR-mediated signaling]. AB - In recent years, major progress has been obtained in studying the early events in TCR-mediated signaling. c-Cb1 has been found to be a negative regulatory factor of the tyrosine kinases in ZAP-70/SyK family. The studies on LAT, SLP-76, ItK and Vav have shown their roles in the signal transduction of Ras and phospholipase Cx1 to Ca2+. Micro-glycolipid raft also plays important role in T cell activation. This minireview shows a brief introduction to the process of TCR mediated signaling. PMID- 12905782 TI - [Progress in the study on the molecules in CD28 family]. AB - CD28 family consists of CD28, ICOS, CTLA-4 and PD-1 molecules. The former two are activation receptors and the later two are inhibition receptors. They produce co stimulatory signals combining with the relevant molecules in B7 family, which plays important role in T cell activation and homeostasis among T subsets. Although the mechanism of signaling by CD28 and CTLA-4 has been well studied, many questions still remain to be answered. Further investigations are required for substantiating the dual-signaling model. PMID- 12905783 TI - [Discovery of MHC restriction in antigen recognition of T cells]. AB - T cell can only recognize specific antigenic peptide-MHC complex on antigen presenting cell. This is MHC restriction in antigen recognition of T cell. This phenomenon was discovered by an Austrian scientist Peter. C. Doherty and a Swedish scientist Rolf. M. Zinkernagel by chance. Then, in order to explain the phenomenon, they proposed two hypotheses: dual receptor and modified self. In the during following 20 years numbers of scientists spent great amount of time in the study of the phenomenon. The process of cell-mediated immune response becomes clear, which greatly promotes the advancing of immunology and many related disciplines. PMID- 12905784 TI - [The position and application of nanotechnology in the development of life science]. AB - In this article, the position of nanotechnology and its effects on life science are concisely discussed, including the effects of nanotechnology on biology, on the study of new type biomaterials, on the development of new drugs, on diagnosis and therapy methods. The importance of nanotechnology in life science is summarized, and the application and development of nanotechnology in life science are also discussed. PMID- 12905785 TI - [Studies on the blood compatibility of the polyurethane/nano-sized carbon composite]. AB - OBJECTIVE: To investigate the effects of nano-sized carbon as a dispersed phase on blood compatibility of polyurethanes. METHODS: A novel nanoscale polymeric composite film was prepared by dispersing nano-sized carbon fiber (vapor growth carbon fiber) into the polyurethane solutions. The surface blood compatibilities of the composites were analyzed and evaluated through platelet adhesion measurement using epifluorescent video microscopy and the variation of fibrinogen and free hemoglobin concentration in the blood contacting the composite respectively. RESULTS: It was showed that the platelet adhesions were highly suppressed on the composite surfaces pre-adsorbed or non-pre-adsorbed with fibrinogen. The changes of the concentration for both free hemoglobin and fibrinogen in the blood contacting the composite surface in the circulations were less than the ones contacting the reference surface. CONCLUSIONS: Introducing nano-sized carbon into the polyurethane matrix showed an improvement of antithrombogenicity for the polyurethane materials. It might be a new promising way to develop biomaterials with good blood compatibility. PMID- 12905786 TI - [Preparation of surface functional magnetic microspheres and their application in nucleic acid separation and enzyme immobilization]. AB - OBJECTIVE: To Prepare surface functional magnetic microspheres for the separation of vascular endothelial growth factor (VEGF) nucleic acid and lactase enzyme immobilization. METHODS: Using suspension polymerization methods to copolymerize MA-styrene containing magnetite nanoparticles and GMA-styrene also containing magnetite nanoparticles, respectively. Both the carboxyl-modified magnetic microspheres and epoxy-modified magnetic microspheres were obtained. In addition, the chloromethyl-modified magnetic microspheres were prepared by seedy microemulsion. The magnetic microspheres bound with b-gamma IgG were determined by radioimmunoassay (RIA), and the separation of VEGF nucleic acid and lactase enzyme immobilization were performed by carboxyl-modified magnetic microspheres. RESULTS: Transmission electron microscopy (TEM), energy-dispersive spectroscopy (EDS) and infrared (IR) spectra showed that the products of polymer magnetic microspheres were monodispersed and that the magnetic particles were uniformly distributed in the microsphere with special functional group on the surface of the microsphere. RIA showed that three kinds of magnetic microspheres could be bound with b-gamma IgG and the absorption of b-gamma IgG reached 75 micrograms/mg, especially for the carboxyl-modified magnetic microspheres. The carboxyl-modified magnetic microspheres can be used for the separation of VEGF nucleic acid by coupling with corresponding primer. Moreover, the immobilized enzyme was proportional to the amount of the carboxyl-modified magnetic microspheres. CONCLUSIONS: The surface functional magnetic polymer microspheres can be bound with active bio-substance, and have a wide application prospect in the fields of biology and medicine. PMID- 12905787 TI - [Nano-hydroxyapatite/collagen composite for bone repair]. AB - OBJECTIVE: To develop nano-hydroxyapatite/collagen (NHAC) composite and test its ability in bone repairing. METHODS: NHAC composite was developed by biomimetic method. RESULTS: The composite showed some features of natural bone in both composition and microstructure. The minerals could contribute to 50% by weight of the composites in sheet form. The inorganic phase in the composite was carbonate substituted hydroxyapatite (HA) with low crystallinity and nanometer size. HA precipitates were uniformly distributed on the type I collagen matrix without preferential orientation. The composite exhibited an isotropic mechanical behavior. However, the resistance of the composite to localized pressure could reach the lower limit of that of femur compacta. The tissue response to the NHAC composite implanted in marrow cavity was investigated. Knoop micro-hardness test was performed to compare the mechanical behavior of the composite and bone. At the interface of the implant and marrow tissue, solution-mediated dissolution and macrophage-mediated resorption led to the degradation of the composite, followed by interfacial bone formation by osteoblasts. The process of implant degradation and bone substitution was reminiscent of bone remodeling. CONCLUSION: The composite can be incorporated into bone metabolism instead of being a permanent implant. PMID- 12905788 TI - [Synthesis of nanometer hydroxyapatite by sol-gel method]. AB - OBJECTIVE: To synthesize nanometer hydroxyapatite (HAP) by sol-gel method from Ca (NO3)2.4H2O and PO (CH3O)3 with water and ethanol as solvents and to study the factors that can affect the formation of sol-gel, including the type of solvents, amount of solvents, pH value, as well as properties under the condition of low temperature combustion of dried gel. METHODS: The viscosities of sol were determined on WU viscosimeter. TG-DTA was used to analyze the thermal procedure of dried gel by increasing temperature. Characterization of the dried gel and powder after combustion were evaluated by X-ray diffraction and infrared spectrum. The morphology of the powder was observed by transmission electron microscopy. RESULTS: Stable and homogeneous gel could be obtained at appropriate pH value when ethylene glycol was added into solvent. The dried gel had the feature of low temperature combustion. HAP powder with particle size of 50 nm was directly prepared. CONCLUSION: Nanometer HAP powder with uniform particle size can be synthesized by sol-gel method. PMID- 12905789 TI - [Preliminary studies on the mechanisms of a new anti-tumor agent PH II-7 with special preference to multidrug resistant tumor cells]. AB - OBJECTIVE: To determine the anti-tumor activity of PH II-7 in vitro and explore preliminarily its mechanisms. METHODS: The anti-tumor activity was measured using colorimetric MTT assay. Apoptosis was determined with fluorescence-activated cell sorter (FACS), electron microscopy and agarose gel electrophoresis. The expressions of mdr1 and sorcin genes were determined by Northern blot assay. RESULTS: PH II-7 inhibited the proliferation of various human tumor cells derived from different tumor cell lines. The IC50 values varied from 0.34-18.61 mumol/L. Especially, PH II-7 had strong inhibitory effect on multidrug resistant tumor cells, whereas adriamycin (ADR) was resistant. Apoptosis was induced in HL60 and HL60/ADR cells treated with 1 microgram/ml PH II-7, while PH II-7 inhibited the expressions of mdr1 and sorcin genes. CONCLUSIONS: PH II-7 is a new potential agent which has strong inhibitory effect on both multidrug resistant cells and their parental cells. PH II-7 may increase the intracellular drug concentration in MDR cells by inhibiting the expressions of the MDR-related genes mdr1 and sorcin and induce the apoptosis of MDR cells and their parental tumor cells. PMID- 12905790 TI - [The transcriptional regulation study on human delta globin gene with CAAT box C- >T point mutation in its promoter]. AB - OBJECTIVE: To study the transcriptional regulation of human delta globin gene with C-->T point mutation at -64 in its promoter. METHODS: Human delta globin genes including wild CAAT box and mutant CAAT box (-64C-->T) were separately cloned into eukaryotic expression vector pcDNA3.1 (-)/Myc-His A which was cut out the strong promoter CMV, transfected MEL cells, and induced by DMSO to express. The transcriptional regulation of human delta globin gene was analysed using semi quantitative RT-PCR. RESULTS: The expression level of human delta globin gene with mutant CAAT box was 2.2-fold as high as that with wild CAAT box. CONCLUSION: The defective CAAT box of human delta globin gene promoter region may be one of the major reasons for its low expression level. PMID- 12905791 TI - [Structural mechanism studies of hTNF alpha mutants in position 30 and 42 amino acid]. AB - OBJECTIVE: To study the relationship between the structure and functional activity of hTNF alpha. METHODS: Four hTNF alpha mutants were constructed, different binding structures and gene responses related with these mutants were studied by the methods of immunoprecipitation and mRNA differential display. RESULTS: The specific activities and LD50 of the different hTNF alpha mutants indicated their different bioactivities. It was shown that the hTNF alpha mutants had the relative binding affinities to the wild types. The mRNA differential display assay proved that the hTNF alpha mutants stimulated different gene responses. CONCLUSION: These results suggest that the specific anti-tumor activities of hTNF alpha mutants are accomplished by inducing different or same gene response at different quantities after its binding to specific receptor. PMID- 12905792 TI - [Analysis of lecithin-cholesterol acyltransferase cDNA and protein sequence from tree shrew]. AB - OBJECTIVE: To acquire cDNA sequence of lecithin-cholesterol acyltransferase (LCAT) from tree shrew and analyze the sequence structure. METHODS: The first strand cDNA was acquired by reverse transcription using mRNA from tree shrew liver as template. By the method of SMART RACE PCR, tree shrew LCAT cDNA was acquired and deduced its amino acids sequence. The sequence and structure of tree shrew LCAT cDNA and amino acid were analyzed and predicted by the molecular software. RESULTS: Tree shrew LCAT cDNA is composed of 1,340 bp, including 2 bp 5' untranslated region (5' UTR), 1,320 bp open reading frame (ORF) which encodes protein precursor of 440 amino acids (24 amino acids signal peptide and 416 amino acids mature peptide), and 18 bp 3' untranslated region (3'UTR). The stop codon is TAA and there is a poly (A) signal sequence AATAAA and a 25 bp poly (A) tail. Tree shrew LCAT cDNA sequence has been accepted by GenBank as a new gene, accession number AF272861 and its homology with human and baboon was 90% and 89%, respectively. CONCLUSION: The sequence of LCAT cDNA in tree shrew has high identity with that of human and other experimental animal species. PMID- 12905794 TI - [Effect of platelet factor 4 on the adhesive property of leukemic CD34+ KG1a cell]. AB - OBJECTIVE: To study the effect of PF4 on the adherence of leukemic CD34+ KG1a cell to human umbilical vein endothelial cell line ECV-304 cell and on the expression of adhesive molecules. METHODS: Adhesion assay and adhesion blocking assay were respectively applied to measure the effect of PF4 and/or adhesion molecule monoclonal antibodies on the adhesion property of KG1a. The expressions of adhesion molecules were determined by RT-PCR and FACS analysis. RESULTS: The adhesion of KG1a cells to ECV-304 was significantly enhanced in the presence of PF4. Such enhancement was also observed when KG1a or ECV-304 cells were separately treated with PF4 before interaction. The adhesion capacity of KG1a cells was reduced when cells were co-incubated with the blocking monoclonal antibodies (MoAbs) against CD49d, CD106, CD54, respectively. In contrast, MoAbs against CD62L, CD62P and CD62E had no such effect. During a period of 3 hours when KG1a or ECV-304 cells were respectively incubated with PF4, the mRNA expressions of CD49 d, CD54 were up-regulated. Furthermore, when KG1a or ECV-304 cells were incubated with PF4 for 2 hours, respectively, the percentages of CD49d+ KG1a cells and CD54+ ECV-304 were increased significantly. CONCLUSION: PF4 can enhance KG1a cell adhesive capacity by increasing the expressions of adhesion molecules. PMID- 12905793 TI - [Effects of TNF-alpha or mmLDL on PAI-1 expression in human umbilical vein endothelial cells]. AB - OBJECTIVE: To demonstrate the changes of PAI-1 activity and mRNA expressions induced by tumor necrosis factor alpha (TNF-alpha) or minimally modified low density lipoprotein (mmLDL) in human umbilical vein endothelial cells (HUVECs). METHODS: The PAI-1 activity in HUVECs culture medium was measured by chromogenic assay. The PAI-1 mRNA expressions were determined by Northern blot. The mitogen activated protein kinase-kinase (MAPKK) inhibitor and protein kinase C (PKC) inhibitor were used to intervene the inductions by TNF-alpha or mmLDL. RESULTS: The PAI-1 activity and mRNA level were increased when HUVECs were exposed to TNF alpha or mmLDL. The MAPKK inhibitor (PD98059, 60 mumol/L) could markedly inhibit PAI-1 activity and mRNA expression induced by TNF-alpha (100 U/ml) or mmLDL (50 micrograms/ml). But Staurosporine (10 nmol/L) and H7 (15 mumol/L), which were the PKC inhibitors, had no significant inhibition effect. CONCLUSIONS: (1) TNF-alpha or mmLDL could induce PAI-1 activity and mRNA expression in HUVECs. (2) Increase of PAI-1 activity induced by TNF-alpha and mmLDL was related to its mRNA expression. (3) The MAPK pathway may play a role in PAI-1 expression induced by TNF-alpha or mmLDL. PMID- 12905795 TI - [Alterations of DPC4/SMAD4/MADH4 gene detected in paraffin-embedded tissues of human pancreatic carcinomas]. AB - OBJECTIVE: To demonstrate the alterations of DPC4/SMAD4/MADH4 gene in paraffin embedded tissues of pancreatic carcinomas. METHODS: Forty-six cases of resected specimens containing carcinomatous tissue and normal pancreatic tissue were analysed for possible DPC4 gene mutations by polymerase chain reaction (PCR)and single-strand conformation polymorphism (SSCP). The DNA sequencing technique was applied to determine the patterns of gene mutation in the PCR-SSCP positive cases. Fifty-six cases of pancreatic carcinoma along with the specimens corresponding normal pancreatic tissues were studied by in situ hybridization (ISH) and immunohistochemistry (IHC) techniques for gene expression in mRNA and protein level. RESULTS: The homozygous deletion rate of exon 1, 2, 3, 4, 8, 11 of DPC4 gene in pancreatic carcinoma was 28.26%, while the mutation rate of DPC4 gene was 21.74%. In these tumors, there were 3 cases of nonsense mutation, 5 cases of missense mutation, 1 case of deletion and missense mutation, 1 case of insertion mutation. Positive rates of SMAD4 in carcinomatous tissues detected by the ISH and IHC were 53.57% and 58.93% respectively, whereas they were 91.07% and 89.29% in the matched normal tissue respectively. There were significant difference between cancer and normal tissue (P < 0.05). Thrity-two cases were positive of DPC4/SMAD4 with all methods mentioned above, the coincident rate was 87.50% (28/32). The coincidence between gene detection and ISH of SMAD4 was 87.50%, and it was 96.88% between gene detection and IHC of SMAD4. Of all 56 cases, the coincidence of the positive rates of SMAD4 detected by ISH and IHC was 91.07%. No significant difference among the positive rates of DPC4/SMAD4 as detected by the three different techniques (P > 0.05). CONCLUSIONS: The main mechanisms of inactivation of DPC4 gene in pancreatic carcinoma are homozygous deletion and mutation. The product of DPC4 expression is significantly decreased in cancer group compared with the normal tissues. As a tumor suppressor gene, DPC4 alteration is an important molecular event in pancreatic carcinoma, and probably plays a crucial role in cancer development and progression. PMID- 12905796 TI - [Mutations in the D-loop region of mitochondrial DNA in ovarian tumors]. AB - OBJECTIVE: To investigate mutations in the D-loop region of mitochondrial DNA in ovarian tumors. METHODS: The D-loop region of 25 epithelial ovarian tumors together with the adjacent normal tissues were amplified by PCR and sequenced. RESULTS: Among the 25 ovarian tumors, 26 mutations were identified with the mutation rate of 32%. 19 mutations were detected in two cases of borderline carcinoma which was a special type of epithelial ovarian carcinoma. There were 6 microsatellite instabilities among the mutations and 11 new polymorphisms which were not reported previously in the GenBank. CONCLUSIONS: The D-loop region of mitochondrial DNA is a highly polymorphoric and mutable region and the mutation rate is relatively high in patients with ovarian tumors. PMID- 12905797 TI - [Patellofemoral replacement for severe patellofemoral osteoarthritis: a 2-10 years follow-up study]. AB - OBJECTIVE: To evaluate the therapeutic effects of patellofemoral replacement for severe patellofemoral osteoarthritis. METHODS: Patellofemoral replacement was performed in 30 patients (36 knee joints) with severe patellofemoral osteoarthritis, among whom were 5 males and 25 females. The patients ranged in age from 36 to 74 years old (mean +/- s, 55.8 +/- 8.5) and were followed up for 2 10 years. RESULTS: Significant improvement was made in both the knee score and functional score after the patellofemoral replacement. The effective rate was 88.9% and 91.7% evaluated with knee score and functional score respectively. CONCLUSION: Patellofemoral replacement is effective in treating severe patellofemoral osteoarthritis in appropriate cases. PMID- 12905799 TI - [The role of nutritional status on serum immunoglobulins, body weight and postoperative infectious-related complications in patients with Crohn's disease receiving perioperative parenteral nutrition]. AB - OBJECTIVE: To evaluate the role of nutritional status on serum immunoglobulins, body weight and postoperative infectious-related complications in patients with Crohn's disease receiving perioperative parenteral nutrition (PN). METHODS: 32 patients with Crohn's disease receiving perioperative parenteral nutrition in our department between 1984 and 1994 were enrolled in this survey. 16 patients with loss of body weight in the range of 15%-30% were assigned to the malnutrition group, the other 16 patients with normal weight or loss of body weight less than 15% to the control group. Serum IgM, IgG and IgA levels were measured before and after PN by enzyme-linked immunosorbent assays. Liver function, body weight changes and postoperative complications were also analyzed. RESULTS: IgM levels were elevated before PN in both groups [control group: (133 +/- 16) mg/dl, malnutrition group: (139 +/- 41) mg/dl; normal value: (110 +/- 35) mg/dl; P = 0.04], decreased to normal value [(105 +/- 29) mg/dl, P = 0.02] in the malnutrition group while having no obvious changes in the control group [(129 +/- 13) mg/dl, P = 0.34]. No significant changes in concentrations of IgG and IgA were found (P in the range of 0.20-0.57). The average weight gain was 1.862 kg in malnutrition group [before PN: (45.8 +/- 8.9) kg, after PN: (48.0 +/- 8.8) kg; P = 0.005] and no significant changes in the control group [before PN: (55.6 +/- 6.1) kg, after PN: (56.3 +/- 6.0) kg; P = 0.46]. There was an increase in infectious complications in the control group (control group: 4 cases, 25%, malnourished group: 2 cases, 12.5%; P = 0.13). CONCLUSIONS: Perioperative parenteral nutrition ameliorated the humoral immunity, increased the body weight in patients with obvious malnutrition, whereas it had little value for those without or with mild malnutrition. PMID- 12905798 TI - [Clinical features of gastroesophageal reflux disease in geriatric patients]. AB - OBJECTIVE: To evaluate the features of gastroesophageal reflux disease (GERD) in elderly patients. METHODS: 72 patients with GERD were investigated for the history of illness and the results of gastroscopy and 24 hours esophageal bile monitoring of 54 patients were simultaneously randomized to undergo ambulatory pH monitoring. The degrees of esophagitis were graded according to endoscopic findings. RESULTS: Obesity was found in 48% of the elderly group (more than 65 years old) and 49% of the control group (less than 65 years old). There were longer acid and bile reflux time and higher frequency (65%) of hiatal hernia in the elderly, and more patients (35%) had complicated severe grade esophagitis. There were longer time of bile reflux and the higher incidence (76%) of both acid and bile reflux in elderly group than in control group (P < 0.05). But the acid reflux time (%) was similar in two groups (P > 0.05). CONCLUSIONS: The GERD in elderly patients may be associated with obesity and hiatal hernia. The features of elderly GERD patients are high frequency of erosive esophagitis, and high frequency of both acid and bile reflux, as well as longer history of bile reflux time. PMID- 12905800 TI - [Assay of carnitine in plasma and urine of healthy adults]. AB - OBJECTIVE: To determine the levels of carnitine in plasma and daily excretion of carnitine in urine of healthy adults so as to provide the reference standard for studying the changes of carnitine in patients. METHODS: Carnitine in plasma and urine was assayed with high performance liquid chromatography (HPLC). The levels of total carnitine (TC), free carnitine (FC) and acetyl-carnitine (AC) in fasting plasma and the daily excretion of TC, FC and AC in urine were assayed in 40 healthy adults (20 men and 20 women) with standard diet. RESULTS: Good linearity (r 2 > or = 0.999) was observed in assaying TC, FC and AC. The relative standard deviation (RSD) was lower than 9.1% and bias lower than 5.6%. It was showed that the plasmatic levels of TC, FC and AC in healthy men [(53.1 +/- 8.5), (41.2 +/- 6.1), (6.2 +/- 0.6) mumol/L] were significantly higher than those in healthy women [(45.4 +/- 5.6), (35.2 +/- 4.9), (5.7 +/- 0.7) mumol/L] (P = 0.002, 0.002, 0.035). The daily urinary excretion of TC, FC and AC in healthy men [(386.1 +/- 22.9), (180.5 +/- 31.8), (33.8 +/- 3.3) mumol] were also significantly higher than those in healthy women [(240.1 +/- 35.6), (112.7 +/- 22.6), (29.3 +/- 4.3) mumol] (P < 0.0005, < 0.0005, < 0.0005) when the adults were given standard diet. Both the plasmatic levels and the daily urinary excretion of TC, FC and AC were of significantly positive correlation with lean body mass (LBM) (r = 0.501 0.856). The (TC-FC)/FC ratios in plasma were 0.29 +/- 0.05 for male and 0.29 +/- 0.04 for female. CONCLUSION: Good precision and accuracy were observed in assaying carnitine with HPLC. After standard diet, both the level of carnitine in fasting plasma and the daily urinary carnitine excretion of healthy adults were positively correlated with LBM. PMID- 12905801 TI - [Telomerase expression in various lesions of adrenal cortex]. AB - OBJECTIVE: To investigate the expression of telomerase in various lesions of adrenal cortex. METHODS: By autoradiography-based telomeric repeat amplification protocol, telomerase expression was detected in 36 samples of adrenocortical lesions, including 29 cases adrenocortical adenoma (8 Cushing's syndrome, 17 aldosteronism and 4 nonfunctional adenomas), 5 cases of hyperplasia of adrenal cortex (presented with Chushing' syndrome), 2 cases adrenocortical carcinoma, and 4 samples of normal adrenal cortex. RESULTS: Of the 40 samples, 2 cases of adrenocortical carcinomas had telomerase expression, and the others had no telomerase expression detected. CONCLUSIONS: No significant telomerase expression was found among different endocrine functional benign adrenocortical lesions. Telomerase expression may be used as an important marker of malignant adrenocortical tumor. PMID- 12905802 TI - [The relationship between intracellular chloride concentration and ischemia reperfusion-induced arrhythmias in myocardial cells]. AB - We investigated the effects of simulated ischemia on intracellular Cl- concentration ([Cl-]i) in guinea pig ventricular myocardial cells and possible role of the [Cl-]i on the ischemia/reperfusion-induced arrhythmias in perfused rat hearts. Our results provided direct evidence that the [Cl-]i in ventricular muscle was increased under ischemic conditions, which suggested that activation of the Cl-(-)HCO3- exchanger by ischemia would partially contribute to the elevation of [Cl-]i. Application of stilbene derivatives or lowering Cl- concentration in perfusion solution delayed the onset of ischemia-induced deterioration in action potentials, pHi, [Cl-]i, and suppressed the incidence of ischemia/reperfusion-induced arrhythmias. The conclusion was made to emphasize the important role of intracellular Cl- homeostasis in cardiac physiology and pathogenesis of myocardial ischemia/reperfusion injury. PMID- 12905803 TI - [The application of nanotechnology in biomedicine]. AB - Nanotechnology is a new comprehensive system, which prepares substances or/material at molecular or nuclear level in nanometer scale, and nano materials can be explored to apply on broad aspects of industrial engineering sciences. This paper reviews the following contents: developing molecular engineering, biological molecular robot and nanometer information processing system, the application of nanobiomaterials in the field of tissue engineering, constructing drug carriers with nanobiomaterials, developing nanotechnology relating to diagnosis and monitoring. Developmental trend of nanotechnology in biomedical science is discussed prospectively. PMID- 12905804 TI - [Nano biomedical material and its application]. AB - Nano biomedical material is a new area that shows promising prospect. In this paper, the researches and applications of nano inorganic biomaterial, nano polymer biomaterial and nano composite biomaterial are reviewed. The developmental tendency of nano biomedical material is also forecasted. PMID- 12905805 TI - [Nano-structured scaffold materials used in tissue engineering]. AB - Micro- and nano-structured surfaces of scaffold materials have important effects on cells' adhesion and proliferation. Nano-structured surfaces can improve cells' adhesion and biocompatibility of materials. The effects of nano-biomaterials on the development of tissue engineering and the methods of preparation of nano biomaterials such as molecular self-assemble and template technology are discussed. PMID- 12905806 TI - [The use of muscle autograft denatured by microwave for repair of gap in removal of facial neuroma]. AB - OBJECTIVE: To study the clinical value of muscle autograft denatured by microwave for repair of gap in removal of facial neuroma. METHODS: A case of patient with a larger facial nerve Schwann cell neuroma was reported. Based on the good results of our experimental research on rats recently, we completed the operation by transmastoid approach for removal of facial neuroma, and the gap of the nerve was repaired with muscle autograft denatured by microwave. RESULTS: The patient was followed up for one year, and her facial function recovery on the affected side was satisfactory. CONCLUSION: Muscle autograft denatured by microwave technique is convenient, highly efficient for repairing facial nerve gap after removal of facial neuroma. PMID- 12905807 TI - [The mouse model and human disease]. AB - The mouse is an ideal model organism for studies of human disease, because mouse is physiologically very similar to human. Also, there is a large genetic reservoir of potential models of human diseases that has been generated. In addition, high-resolution genetic and physical linkage maps are now available and the sequence of mouse genome will be completed in the near future. Furthermore, the techniques necessary for the modification of mouse genome, such as transgenic and knockout techniques, and chromosome engineering methods have been established. These techniques enable us to introduce any mutations anywhere in the mouse genome. The methods for analyzing complex genetic diseases also have been developed. These advances facilitate the identification and cloning of mouse disease loci and the establishment of new models. It makes mouse the model organism of choice by academic and industrial researchers to study human diseases. In Part I of this review, we summarize the classical and modern approaches that provide the basis of establishing mouse model of human diseases. In the following parts, we will list more than 100 mouse models of human diseases. In most of these models, the mouse mutant phenotype closely resembles the human disease phenotype. These mouse models are valuable sources for the understanding of the human diseases and they can be used to develop strategies for prevention and treatment of the diseases. PMID- 12905808 TI - [The significance of secondary prevention of cancer in China]. AB - During past twenty years (1973-1992), the cancer mortality was increasing significantly in China. For cancer prevention and control nationwide, the primary prevention is important. Pity is that it usually takes longer time to decrease cancer mortality and liable to benefit younger generation. In practice, the secondary prevention, which put emphasis on the massive screening, early detection and early treatment, would decrease the cancer mortality as well as cancer incidences in relatively short time. Therefore, it would contribute much to the local social and economic development and improve the health insurance. In the high risk areas of cancers, such as cervical, esophageal, gastric, liver and nasopharyngeal cancers, the secondary prevention could be carried out as more effective strategy for cancer prevention in China. PMID- 12905809 TI - [The trends and preventive strategies of esophageal cancer in high-risk areas of Taihang Mountains, China]. AB - OBJECTIVE: To analyze and predict the trends of incidence and mortality rates of esophageal cancer in areas of Taihang Mountains, and explore the strategies for facilitating the reduction of the incidence and mortality of esophageal cancer. METHODS: The data were collected from Linxian and Cixian Counties registries between 1988-1997. The trends of the incidence and mortality rates for esophageal cancer had been fitted and predicted by the model of BP (Back Propagation) Neural Networks. Then Linqi Xiang in Linxian County was taken as an example, to determine the power for reducing incidence and mortality of esophageal cancer using endoscopic technology as an early detection and treatment method. RESULTS: There were slowly declining trends for both incidence and mortality rates during past 10 years in Taihang Mountains. The predicting rates of the esophageal cancer incidence in 2,002 are 115. 70/100,000 for male and 79.88/100,000 for female; mortality rates are 94.00/100,000 and 56.29/100,000 respectively. Both incidence and mortality rates a still in high level. But when using endoscopic technology as an early detection and treatment method, the incidence rate is nearly 1/3 of a control group and the power is 88.30% at the end of eight years. The mortality rate is 1/4 of the control group and power is 89.44% after five years intervention (both powers based on 0.05 significant levels). CONCLUSIONS: The incidence and mortality rates for esophageal cancer are still on high levels in Taihang Mountains. We should continue to enhance our preventive effort there. After using endoscopic technology as an early detection and treatment method, the incidence and mortality rates will be decreased markedly during 5 to 8 years. It is recommended to implement a large-scale secondary prevention strategy in these areas. PMID- 12905811 TI - [Clinical characteristics of Parkinson's disease in natural population]. AB - OBJECTIVE: To study the Clinical characteristics of Parkinson's disease(PD) in natural population. METHODS: All the patients diagnosed as Parkinsonism from a prevalence study in Beijing in 1997 were collected for investigation. The spectrum of the clinical features of Parkinsonism and the onset, progression and prognosis of PD were analysed, as well as the impact factors to the prognosis were analysed. RESULTS: 109 cases were ascertained as PD from the sample population, in which 64(58.7%) were PD, 11 (10.1%) were Parkinson plus syndrome and 34(31.2%) were symptom Parkinsonism. 30 of the 64 PD cases were male. The median age of PD was 74.5 years. The median age at onset was 68 years old. The median duration was 5 years. The average Hoehn-Yahr staging score was 2.81. The clinical characteristics of PD and other Parkinsonism were compared, as asymmetry of symptoms and signs was an important feature to differentiate PD from other Parkinsonism. 20.3 percent of PD cases were associated with dementia. 16(25%) of the 64 PD cases initiated with essential tremor and developed to PD during 6-49 years after the onset of tremor. Tremor was the dominant initial symptoms of PD(60.9%). Type of initial symptoms was a relatively definite impact factor to the prognosis of PD. Levodopa treatment was likely to delay the duration of PD. CONCLUSIONS: The clinical features of Parkinsonism in natural population were different as compared with previous studies. A further study on the natural history of PD thoroughly is needed. PMID- 12905810 TI - [Effects of 8 years community intervention on risk factors of cardiovascular diseases in Fangshan Beijing]. AB - OBJECTIVE: To study the changes of risk factors of cardiovascular disease in the rural community population after intervention. METHODS: The Beijing Fangshan cardiovascular prevention program was a community-based comprehensive intervention study which was launched from 1991 and ended in 2000 in five communities including three as intervention communities (IC) and two as control communities (CC) in Fangshan, Beijing suburb. The intervention measures were focused on health education and hypertension control. The changes of risk factors of cardiovascular disease in IC and CC were analyzed using random sample in the year 1991, 1995 and 1999, respectively. The risk factors include systolic and diastolic blood pressure (SBP and DBP), body mass index(BMI), serum total cholesterol(TC), triglyceride(TG), high-density lipoprotein cholesterol(HDL), smoking, and drinking. RESULTS: From the year 1991 to 1999, the risk factors of cardiovascular disease such as SBP, DBP, smoking rate and drinking rate were reduced in the population of IC. For male in IC, the decline of SBP, DBP, smoking rate, and drinking rate were 1.6 mmHg, 1.1 mmHg, 14.5% (P < 0.01) and 3.7%, respectively. For female of IC, SBP and DBP declined 4.8 mmHg (P < 0.01) and 3.2 mmHg (P < 0.01), respectively. SBP, DBP and smoking rate in the population of CC had a little reduction while BMI, TC and TG increased in both IC and CC. During the period of 1991 to 1999, most cardiovascular risk factors in the population of IC had net reduction compared to that of CC. CONCLUSIONS: Except for BMI and lipids, rural community intervention, as focused on health education and hypertension control, has resulted in the reduction of most risk factors of cardiovascular disease. PMID- 12905812 TI - [Relation of hypertension to diabetic nephropathy in patients with non-insulin dependent diabetes mellitus--a pair-matched case-control study]. AB - OBJECTIVE: The study aimed to assess the role of hypertension in the cause of nephropathy in patients with NIDDM. METHODS: A retrospective analysis was done on two groups of non-insulin-dependent diabetes mellitus (NIDDM) patients, one group without proteinuria (< 300 mg/24 h, n = 106) and the other group with proteinuria (> or = 500 mg/24 h, n = 106). The groups were matched by age (< or = +/- 3) years, sex, race, and resident place. Some information of these subjects including demographic, history of disease, family history of diseases, life and behavior style variables were obtained by questionnaire, some variables were measured including systolic blood pressure(SBP), diastolic blood pressure(DBP), fasting blood glucose, and quantity of protein in 24 h urine. Finally the conditional logistic regression analysis was done. RESULTS: Some factors were independently associated with the occurrence risk of diabetic nephropathy (DN) associated history of hypertension, longer duration of hypertension, higher levels of the past highest SBP and DBP. Their corresponding odd ratios(OR) with 95% confidence intervals(CI) were 2.00(1.17-3.43), 1.25(1.08-1.46), 1.38(1.15 1.66), and 1.33(1.09-1.62) respectively. But family history of hypertension was not significantly associated with the development of DN. When the above-mentioned relations were adjusted by some relevant confounding factors, the associations were still present. CONCLUSIONS: History and longer duration of hypertension, history of the highest SBP and DBP are independent risk factors for DN. PMID- 12905813 TI - [Cloning of human B lymphocyte activation-related novel gene]. AB - OBJECTIVE: To clone the novel activation-related gene of B lymphocyte. METHODS: The differential display reversal transcription PCR (DDRT-PCR) technique was applied to analyse the expression difference of mRNA between resting and activated B lymphocyte from human tonsil. The positive differential display cDNA fragment identified by Northern-blotting was chosen as probe to filtrate human activated B lymphocyte cDNA library. RESULTS: Sixty two differential display cDNA fragments (expressed sequence tag, EST) were obtained. Thirty-two of them were mainly expressed in resting B lymphocyte and thirty were expressed in activated cells. Twenty-five were positive ones after identification by Northern blot analysis. A novel cDNA clone was obtained after using EST30 as a probe to filtrate the human activated B cell cDNA library. The whole cDNA clone was 2,048 bp in length and contains a 630 bp open reading frame. The N end of the deduced amino acid sequence was homologous with KAR3 protein which is a member of kinesins superfamily in yeast. CONCLUSIONS: A novel possible activation-related gene in human B lymphocyte was obtained. PMID- 12905815 TI - [Gene cloning and expression of human soluble interleukin-6 receptor in insect cells]. AB - OBJECTIVE: To clone the human soluble interleukin-6 receptor(hsIL-6R) gene and expression in insect cell line. METHODS: The hsIL-6R gene was cloned into plasmid pAcGP67B. After co-transfection of recombinant plasmid and wild type AcNPV DNA, the rAcNPV was confirmed by the end point dilution assay and dot blot. Then it was purified by plaque assay. RESULTS: SDS-PAGE showed molecular weight of the expressed product was about 47,000. The expressed recombinant protein was confirmed to be specific and capable of binding its ligand IL-6 by Western blot and ligand-receptor binding assays. CONCLUSIONS: Secretory expression of hsIL-6R gene in baculovirus expression system was indicated. The expressed product had immunological and biological activities. PMID- 12905814 TI - [Differentiation and malignant suppression induced by mouse erythroid differentiation and denucleation factor on mouse erythroleukemia cells]. AB - OBJECTIVE: To investigate the roles of mouse erythroid differentiation and denucleation factor (MEDDF), newly cloned in our laboratory, in erythroid terminal differentiation. METHODS: Mouse erythroleukemia cells (MEL) were transfected with eukaryotic expression plasmid pcDNA-MEDDF. The changes of cell growth rate, mitotic index and colony-forming rate in semi-solid medium were investigated. The expressions of c-myc and beta-globin genes were analysed by semi-quantitative RT-PCR. RESULTS: MEL cells transfected with pcDNA-MEDDF showed significant lower growth rate, mitotic index, and colony-forming rate in semisolid medium(P < 0.01). The percentage of benzidine-positive cells was 32.8% after transfection. The expression of beta-globin in cells transfected with pcDNA MEDDF was 3.43 times higher than that of control (MEL transfected with blank vector, pcDNA3.1), and the expression of c-myc was decreased by 66.3%. CONCLUSIONS: MEDDF can induce differentiation of MEL cell, and suppress its malignancy likely. PMID- 12905816 TI - [Development of a canine adenovirus type 1 vaccine strain E3-deleted based expression vector]. AB - OBJECTIVE: To evaluate canine adenovirus type 1 vaccine strain (Cannaught Laboratory Limited, CLL) as recombinant vaccine and gene transfer vector. METHODS: Recombinant virus CLLEGFP which contains enhanced green fluorescent protein(EGFP) reporter gene was constructed. CLLEGFP was used to infect various human derived cell lines(293, Hela, CO, SW, Hep-2 and CAM) by inoculating intraperitoneally(i.p.), intravenously(i.v.) and intramuscularly (i.m.) to Kunming mice other than oral administration. Various tissue samples of the mice were collected at multi-time point for observing EGFP green fluorescence. Anti EGFP antibodies were detected by Western blot analysis in the sera after 4 weeks. RESULTS: CLLEGFP can infect various human derived cell lines and express EGFP. EGFP green fluorescence were observed in liver tissue cells after i.p. transducing 3 days. All immune inoculation ways above could induce Kunming mice producing anti-EGFP antibodies which were identified by Western blot analysis. CONCLUSIONS: These results indicate that CLL possess powerful potential as recombinant vaccine and gene transfer vector. PMID- 12905817 TI - [Polymorphism of parathyroid hormone gene and its association with bone mineral density of Han nationality women in Beijing area]. AB - OBJECTIVE: To investigate the distribution of frequency of parathyroid hormone(PTH) gene polymorphisms in healthy adults of Han nationality in Beijing area and relationship between PTH genotypes and bone mineral density(BMD) in young and postmenopausal women. METHODS: Polymorphism of PTH gene were detected by polymerase chain reaction restriction fragment length polymorphism(PCR-RFLP) of restriction enzyme Bst B1 in 270 subjects. If the site of enzyme Bst B1 existed, the PTH genotype was "B". On the contrary, if base mutation occurred, the genotype was "b". Some of the PTH genotypes were confirmed by DNA sequences analysis. Bone mineral density(BMD) was measured by dual-energy X-ray absorptiometry. RESULTS: Genotype frequencies of BB, Bb, bb were 73.7%, 25.9%, and 0.4% respectively in adults of Beijing areas. The frequencies of RFLP alleles B, b were 86.7% and 13.3%. Beijing postmenopausal women frequencies of BB, Bb, bb were 67.1%, 32.2% and 0.7%. B, b alleles frequencies were 83.2% and 16.8%. We statistically compared bone mineral density at the lumbar 2-4, neck, wards triangle and trochanter major, there was no significant difference between BB and Bb genotype of young women and postmenopausal women groups. Otherwise, no obvious relationship was found between the BMD and PTH genotype in Beijing women. CONCLUSIONS: PTH gene polymorphisms were not associated with BMD in Beijing women. PMID- 12905819 TI - [Influences of age and gender on dose-response and recovery time-course of atracurium]. AB - OBJECTIVES: To evaluate the influences of age and sex on dose-response and recovery time course of atracurium. METHODS: Seventy-two patients (38 men and 34 women), ASA grade I, aged 15-59 years, scheduled for elective surgery were included in the study. All patients were divided into three groups according to their ages: Group 1, 32 patients aged 15-29 years; Group 2, 21 patients aged 30 40 years, and Group 3, 19 patients aged 41-59 years. General anesthesia was maintained with 60% nitrous oxide in oxygen, thiopentone and fentanyl. Neuromuscular function was assessed by train-of-four(TOF) stimulation of accelerometry and the percentage depression of the first response (T1) of the TOF stimulus was used as the study parameter. The dose-response relations of atracurium were determined by the cumulative dose-response technique. RESULTS: As compared to older and female patients, dose-response curves of atracurium in young and male patients were shifted in a parallel fashion to the left. The dose response relation of atracurium correlated closely with patients' ages and sexes by multiple linear regression analysis. There were significant differences of ED50, ED90 and ED95, and clinical duration and total duration of action following administration of atracurium of same doses between the three age groups as well as men and women. CONCLUSIONS: Age and gender could significantly affect the pharmacodynamics of atracurium. PMID- 12905818 TI - [Inhibitory effect of melatonin on the development of pituitary prolactin producing tumors induced by 17-beta-estradiol]. AB - OBJECTIVE: To examine the inhibitory effect of melatonin (MLT) on the development of pituitary prolactin-producing tumors (prolactinoma) induced by 17-beta estradiol (E2), in vivo, and explore MLT's oncostatic mechanisms. METHODS: The prolactinomas were established by implanting E2-laden silastic capsules subcutaneously in Sprague-Dawley male rats. MLT doses 0.05, 0.25, 0.50, 1.00, and 2.00 mg/rat were administrated separately to 5 groups subcutaneously starting seven days prior to tumor induction for 97 days. The matched controls were given equal volumes of 4% alcohol in saline. RESULTS: (1) The prolactinoma weights in 0.05, 0.25, 0.50, 1.00 and 2.00 mg MLT dose groups were 25.91% (P > 0.05), 48.78% (P < 0.01), 36.78% (P < 0.05), 31.04% (P > 0.05) and 35.22% (P > 0.05) respectively which were lower than that of control group; (2) The PRL mRNA levels of prolactinoma in 0.05, 0.25, and 0.50 mg MLT dose groups were 33.67% (P < 0.05), 25.51% (P < 0.05) and 41.84% (P < 0.01) respectively which were lower than that of control group as estimated by Northern Blot, and the in situ hybridization studies; (3) The DNA contents of prolactinoma in 0.05, 0.25 and 0.50 mg MLT dose groups were 40.73% (P < 0.001), 51.15% (P < 0.001) and 60.23% (P < 0.001) respectively which were lower than that of control group by laser scanning confocal microscopy; (4) Plasma peroxidative lipid contents in 0.05, 0.25, 0.50, 1.00 and 2.00 mg MLT dose groups were 26.45% (P < 0.05), 23.97% (P < 0.05), 47.11% (P < 0.001), 66.12%(P < 0.001) and 64.46% (P < 0.001) respectively which were lower than that of control group. The correlation coefficient between MLT doses and plasma peroxidative lipid contents was -0.8257 (P < 0.05). CONCLUSIONS: MLT in suitable doses is able to inhibit the development of E2 induced prolactinoma by inhibiting the expression of PRL gene and the DNA synthesis. The link between MLT antioxidative action and its inhibitory effect on development of prolactinoma should be further investigated. PMID- 12905820 TI - [Relation between alkaline phosphatase in gingival crevicular fluid of implant and the curing result]. AB - OBJECTIVE: To investigate the relation between alkaline phosphatase (ALP) in gingival crevicular fluid (GCF) and prognosis. METHODS: We measured the alkaline phosphatase in gingival crevicular fluid among 56 cases of implant tooth which included 2 failed cases, 5 cases in bad oral hygiene and with gingivitis, compared with the normal group consisted of 10 healthy persons. RESULTS: The difference of ALP level between normal group and success implant group is not significant, but between normal group and success with gingivitis group is significant(P < 0.05). The ALP level of 2 failed cases are highest (because of few failed cases, no statistics was done). CONCLUSIONS: The ALP level in GCF is supposed to be an important index in evaluating the result of the implant. PMID- 12905821 TI - [Masseter thickness measured by ultrasonography of 50 young healthy adults in relation to facial morphology]. AB - OBJECTIVE: To quantify the normal range of the ultrasonically measured thickness of the masseter which is related to the variation of the facial morphology in different young individuals. METHODS: The masseter thickness of 50 young healthy adults (25 men and 25 women) was measured bilaterally under relaxed conditions and with maximal clenching by a real-time ultrasound imaging technique. The relation between masseter thickness and facial morphology was also observed. RESULTS: Under relaxed conditions, the mean thickness of the muscle in men was(11.83 +/- 1.19) mm, and under contracted conditions, (15.51 +/- 1.40) mm. In women, the respective measurements were(9.84 +/- 1.03) mm and (13.02 +/- 1.10) mm. In men the thickness of masseter had a negative linear correlation to the indices of FH/FWa (the ratio of the anterior facial height to the intergonial width) and FWz/FWa (the ratio of the bizygomatic facial width to the intergonial width). CONCLUSIONS: The mean thickness of the masseter in men was larger than that in women, and the thickness of the muscle was related to the male facial morphology. PMID- 12905822 TI - [Fudenine, relative to blood glucose-control, is a novel membrane protein]. AB - OBJECTIVE: To determine whether Fudenine is a novel membrane protein. METHODS: Green fluorescence protein(GFP) was used to localize Fudenine in vivo. GFP, as a control, was targeted to cytoplasm. Epithelial cell, CBRH7919, and non-epithelial cell, L-6TG, were cultured and transiently transfected by using the lipofectamine reagent. After 48 h, intact cells were examined with fluorescence microscope for Fudenine. RESULTS: Reporter plasmid pEGFP-N1, as a control, was expressed and localized to cytoplasm. But Fudenine, driven by the cytomegalovirus promoterenhancer contained in the pEGFP-N1 vector, was overexpressed and targeted to cellular membrane. CONCLUSIONS: Fudenine is a novel membrane protein. It may play the similar role with its homologues AC133 antigen and prominin in human and mouse, respectively. It might be involved in signaling transduction and regulate blood glucose metabolism in vivo. PMID- 12905823 TI - [Cancer risk assessment among medical X-ray workers in China]. AB - Cancer incidence(1950-1995) among 27,011 medical X-ray workers in comparison with 25,782 non X-ray medical specialists between 1950 and 1980 in China was investigated. The average cumulative dose received by the X-ray workers also reconstructed by retrospective dosimetry methods. Significant cancer risk was seen among medical X-ray workers(RR = 1.2). Significantly elevated risks were found in leukemia, cancer of skin, female breast, lung, liver, bladder and esophagus, the RRs were 2.2, 4.1, 1.3, 1.2, 1.2, 1.8 and 2.7 respectively. The patterns of cancer risk were associated with years since beginning of X-ray work, age and calendar year of initial employment and cumulative dose suggest that the risks of leukemia, skin cancer and female breast cancer, possibly thyroid cancer were related to occupational exposure to X-rays. A significant cancer risk could be induced by prolonged exposure to low dose ionizing radiation when the cumulative dose reached a certain level. PMID- 12905824 TI - [30-year experiences on early detection and treatment of esophageal cancer in high risk areas]. AB - The author had conducted the clinical study on early diagnosis and early therapy of esophageal cancer (EC) in the area of high prevalence for more than 30 years. In the endoscopic survey, 742 patients were diagnosed as early EC from 22,285 participants. The endoscopic appearance of the mucosa in early EC was classified into three types: (1) change of mucosa color: red and white area; (2) Increased thickness and change of the vascular pattern of the mucosa; (3) mucosa morphology: erosion, plaque, coarseness and nodular. Iodine staining under endoscopic examination had promoted the early detection of EC and precancerous lesions. Since 307 patients with EC at early stage (65 carcinoma in situ, 95 intramucosal carcinoma, and 147 submucosal carcinoma) had undergone esophagectomy. There were no lymphatic metastasis in carcinoma in situ, however there were 2 cases (2.11%) of intramucosal carcinoma and 27 cases (18.37%) of submucosal carcinoma with lymph node metastasis. 24-year follow-up, results showed that 5-year survival rate is 84.1% and a 10-year survival rate is 72.9% among the cases of esophagectomy. A 5-year survival rate is 100% in 22 cases followed up for full five years among 141 cases with endoscopic mucosectomy. According to this study, severe dysplasia and some moderate dysplasia were precancerous lesions. Should these precancerous cases be treated, the cancerization risk of target population would reduce dramatically and the incidence of EC would be decreased down to 2/3. PMID- 12905825 TI - [Epidemiological studies on the relationship between psychosocial factors and cardiovascular disease]. AB - Psychosocial factors contribute significantly to the pathogenesis and expressions of cardiovascular disease(CVD). Recent studies have demonstrated that 5 specific psychosocial entities are most relevant: (1) depression, (2) anxiety; (3) personality factors and character traits; (4) social isolation, and (5) chronic life stress. Epidemiological studies consistently demonstrate a significant prospective relationship between the occurrence of major depression and the cardiac events. Increasing evidence now links anxiety disorders to the development of CVD events in general populations. Although type A behavior is continuously found to be positively related with increased risk of CVD, a series of studies have reported no correlation between type A behavior and CVD risk. However, hostility, a major attribute of the type A behavior pattern, is considered to be more pathogenic. Social isolation characterized mainly by a relatively small social network has been shown to be associated with increase in the incidence of CVD over time. Finally, chronic life stress, particularly the work-related stress, proved to be closely related to CVD risk. The pathophysiological mechanisms of psychosocial factors on the pathogenesis of CVD are: (1) excessive sympathetic nervous system activation, (2) triggering of myocardial ischemia; (3) promotion of arrhythmogenesis, (4) stimulation of platelet function, and (5) deterioration of endothelial function. PMID- 12905826 TI - [Non-steroidal anti-inflammatory drugs and chemoprevention of digestive cancer]. AB - Recent epidemiology and laboratory studies indicate that regular taking of aspirin and other non-steroidal anti-inflammatory drugs(NSAIDs) may reduce the risk of colorectal, esophageal, stomach and pancreatic cancers and other digestive cancers. Thus, aspirin and other NSAIDs may be an effective chemoprevention agent for digestive cancers. On the other hand, this protection effort may be beneficial to the course of the intervention, regression and prevention of cancer lesions. The possible mechanism of NSAIDs chemoprevention may be: (1) reducing the synthesis of prostaglandin(PG) and inhibiting cyclo oxygenase(COX) activity; (2) inducing apoptosis in epithelial cells of the gastro intestinal origin; (3) obstructing signaling transduction pathways of COX and PG. Now, chemoprevention of NSAIDs has become focus of research on cancer secondary prevention, as its protective effects of chemoprevention of digestive cancer have been determined. NSAIDs, especially selective COX-2 inhibitor may be a novel useful chemoprevention agents for digestive cancer and their precursor lesions in future. PMID- 12905827 TI - [Advances in genetic epidemiology]. AB - We have discussed some advances in genetic epidemiology, which include the effect of gene-environment interaction on diseases. The developing genetic tests and genetic information will benefit the health care and disease prevention. PMID- 12905828 TI - [Statistical approaches to gene mapping of human diseases]. AB - Gene Mapping of human diseases, especially the complex genetic diseases, has been the difficulty and hotspot in medical genetics and gene study. In this article, the principle and application of several gene mapping methods are discussed, including linkage analysis (parametric and nonparametric linkage analysis), association study and linkage disequilibrium analysis. PMID- 12905829 TI - [The nucleic acid analysis by DNA chip technique based on nuclease S1 protection]. AB - OBJECTIVE: Establishing a method for quantitative analysis of nucleic acid by DNA chips. METHODS: The modified oligonucleotides with ribose at 3'-end was chemically synthesized. The 5'-end was labeled by radioisotope 32P with kinase catalyzed reactions. Such oligonucleotides were converted into di-aldehyde at 3' end by oxidization with NaIO4, and then were spotted on glass slide with the amino group modified surface. After reduced with NaBH4, the oligonucleotides were attached strongly. The DNA chips prepared with this method were hybridized with nucleic acids existed in the solution and then digested with nuclease S1. RESULTS: When they were paired with the nucleic acids in the solution perfectly, the oligonucleotides on the chip were not cleaved by nuclease S1. Otherwise, the oligonucleotides on chip were cleaved. The protection efficiencies appeared proportional to the perfect paired nucleic acids in the solution when the content of target nucleic acids were less than the spots on the slides. CONCLUSIONS: The method was developed for both qualitative and quantitative analysis of nucleic acid. As it was not required to label the samples with radioisotope or fluorescence, it might be a practical choice for clinical tests. PMID- 12905830 TI - [Determination of cholesterol in lipoprotein fractions by agarose gel electrophoresis]. AB - OBJECTIVE: To evaluate a single step electrophoresis for quantitative determination of cholesterol of high-, low-, very-low-density lipoprotein(HDL, LDL, VLDL) and fast pre-beta lipoprotein [lipoprotein (a), Lp(a)]. METHODS: Quantification of lipoprotein cholesterol was performed by enzymatic staining of cholesterol in a new agarose gel electrophoresis method that allows the separation of LDL, VLDL, HDL, and Lp(a) by Helena REP system. The results of electrophoresis method were compared with those by traditional method like PTA Mg2+ precipitation method for HDL-C, PVS precipitation method for LDL-C, and Immunoturbidimetric assay(ITA) method for Lp(a). RESULTS: Within-runs CV were 5.16%-7.46%, 1.26%-3.28% and 3.78%-5.86% for VLDL-C, LDL-C and HDL-C, respectively. Between-runs CV were 8.35%-11.25%, 2.78%-4.08% and 4.23%-6.36%, respectively. The linearity of this method was up to 10.35 mmol/L total cholesterol. The recoveries were 90.3%, 94.3% and 89.6%, respectively. No interference were observed when bilirubin(< 342 mumol/L), hemoglobin(< 20 g/L) or triglyceride(< 11.0 mmol/L) were added to pooled serum, respectively. There was good agreement between methods, with r = 0.9557 for HDL-C(electrophoresis method vs PTA-Mg2+ precipitation method), r = 0.9609 for LDL-C(electrophoresis method vs PVS precipitation method) and r = 0.9235 for Lp(a)-C (electrophoresis method) vs Lp(a) (ITA method). CONCLUSIONS: The electrophoresis method offers a simple and inexpensive means of simultaneously measuring HDL-C, VLDL-C, Lp(a)-C and LDL-C. PMID- 12905832 TI - [Effects of anti-ABL tyrosine kinase intrabody on the growth of K562 cells in nude mice]. AB - OBJECTIVE: To study the effects of anti-ABL tyrosine kinase intrabody on the growth of human chronic myelogenous leukemia (CML) cells in nude mice. METHODS: A recombinant retroviral vector MSCV-ibE-IRES-eGFP was constructed to express intracellular single-chain antibody (intrabody) against ABL tyrosine kinase domain in CML cells. K562 cells were transduced with the retrovirus, eGFP+ cells were then selected by fluorescence-activated cell sorting (FACS). The intrabody mRNA expression was determined by reverse transcription (RT)-polymerase chain reaction (PCR). BCR/ABL and c-ABL protein tyrosine kinase (PTK) activity in the cells was examined. Transduced cells and control group K562 cells were transplanted into nude mice respectively and the tumor sizes were dynamically observed. RESULTS: K562-ibE cell was obtained. Expression of the BCR/ABL and c ABL protein tyrosine kinase activity of harvested K562-ibE cells were markedly inhibited. At 14, 21 and 28 days after cell injection, the tumor volumes of experimental mice were obviously smaller than that of control mice, about one half of the control groups (P < 0.05). CONCLUSION: The growth of K562-ibE cells was significantly inhibited in vivo. It is possible that inhibition of the BCR/ABL protein tyrosine kinase activity by the intrabody blocked BCR/ABL signal transduction pathway, promoted apoptosis and reduced tumorigenicity of K562 cells in vivo. PMID- 12905831 TI - [The effects of cytokines mediated ex vivo expansion on the cell adhesion molecule expression of cord blood hematopoietic stem and progenitor cells]. AB - OBJECTIVE: To compare the expression of cell adhesion molecules (CAMs) among VLA 4 (CD49 d), VLA-5 (CD49e), LFA-1 (CD11a), L-selectin (CD62L), and PECAM-1 (CD31) which are more related to the homing of hematopoietic stem and progenitor cells (HSPC) on the ex vivo expanded CD34+ subset with that of fresh isolated AC133+ cells. METHODS: AC133+ cells selected from fresh cord blood (CB) samples were cultured in QBSF-60 serum-free media in the presence of Flt-3 ligand + SCF + TPO (FST), with initial addition of IL-3 for up to 2 week. Expansion potential and the expression of above CAMs were evaluated at day 0, day 7, day 10 and day 14. RESULTS: (1) Simultaneously numerical expansion of various HSPC was constantly observed during the culture, and the fold expansion of AC133+ cells and CD34+ cells on day 14 were 33.50 and 64.56 respectively; (2) The number of CD34+ subsets expressing the above adhesions were all increased at different degrees (from 20 fold to 160 fold). (3) The expressions of CD11a, CD49d, and CD49e on ex vivo expanded CD34+ cells were increased as compared to their baseline levels, but the percentage of CD62L+ and CD31+ subpopulations in CD34+ cells were decreased. CONCLUSIONS: Our short-term culture system can not merely support the simultaneous expansion of CB derived AC133+ cells, but the expanded hematopoietic progenitors may well sustained the expression of homing-related adhesion molecules. PMID- 12905833 TI - [Committed hematopoeitic differentiation of embryonic stem cell]. AB - OBJECTIVE: To explore a stable technology for inducing ES cell to committed hematopoietic differentiation. METHODS: The effects of various inductive factors on BLast Colony-Forming Cell (BL-CFC) were investigated. In vitro differentiative system for ES cells was employed in this study. RESULTS: A high linear correlation between the number of D3.5 EB-derived cells plated and the number of blast cell colonies was developed, r = 0.9931. With high frequency of blast colonies observed (1.08-1.2 colonies per 100 cells). 20%-30% D4T conditioned medium (D4T CM) showed the most significant growth potentials of blast colonies. D4T CM, EPO or KL alone had no blast colony growth promoting effect (P > 0.05). But VEGF alone had high significant blast colony growth promoting effect (P < 0.001). However, any two factors combination from above four factors exerted better growth promoting effect than VEGF alone (EPO + D4T CM, P < 0.05; KL + D4T CM, P < 0.01; VEGF + D4T CM, P < 0.001). There were no significant difference among VEGF + KL and EPO + D4T CM or KL + D4T CM, and KL + D4T CM (P > 0.05). While the combination of VEGF + D4T CM was better than KL + D4T CM, VEGF + KL or EPO + D4T CM (P < 0.001). Moreover, the combination of VEGF + KL + D4T CM + EPO, had the highest significant blast colony growth promoting effect (P < 0.001). And the highest frequency of blast colonies was observed (1.5-1.2 colonies per 100 cells). CONCLUSION: VEGF may be the main factor which stimulates the growth of significant numbers of blast cell colonies. D4T CM maybe contains strong cofactors. EPO and KL are the main factors for the induction of BL-CFC to committed hematopoietic differentiation. D3.5 EB-derived cells are more sensitive to various stimulators and have strong blast colony growth promoting effect than that of D3.25 EB-derived cells. PMID- 12905834 TI - [Transplantation of mesenchymal derived stem cells followed by G-CSF injection can reconstitute hematopoiesis of lethally irradiated BALB/c mice]. AB - OBJECTIVE: To explore the hematopoietic reconstitution potential of mesenchymal derived stem like cells. METHODS: We transplanted bone marrow mesenchymal derived stem like cells into lethally irradiated BALB/c mice. Hematopoietic cells were derived from the non-adherent bone marrow cells 24 hours after initial culture while murine mesenchymal derived stem like cells from bone marrow of donor mice were cultured for 10 days before the transplantation. RESULTS: All mice of group 1 and 3 died in 7-8 days post irradiation following transplantation, while all the mice from group 2 and 4 survived. The time course of hematopoietic reconstitution was then observed. The peripheral blood and bone marrow cell count recovered in the MSC + G-CSF transplanted group and the BM transplanted group after 3 weeks. Interestingly, CFU-GM number in the MSC + G-CSF transplanted group increased significantly after 2 weeks and even more than that in the BM transplanted group after 3 weeks while as CFU-GM colony dropped 2 weeks after in the BM transplanted group. Spleen colony (CFU-S) number and size of the MSC + G CSF transplanted group was significantly greater than the BM transplanted group. Furthermore, PCR analysis was performed using peripheral blood cells to determine if any male-derived cells were present. No male-derived cells were found in any of the mice from group 1 and 3. Y-chromosome-specific src gene was found to be dominant in the MSC + G-CSF transplanted group and the BM transplanted group by week 4 post transplantation. In addition, we demonstrated that induction with G CSF lead to CFU-GM colony formation from MSC compartment in vitro. CONCLUSION: These results indicate that under stimulation of G-CSF, mesenchymal derived stem like cells might differentiate into hematopoietic primitive stem cells in vivo and have the capacity to re-establish hematopoiesis in lethally irradiated mice. This study should provide an alternative transplantation treatment for malignancy. PMID- 12905835 TI - [The CXC-chemokine platelet factor 4 (PF4) increases KG1a cells adherence and modulates actin polymerization of KG1a cells]. AB - OBJECTIVE: To study the effects on adherence of hematopoietic stem/progenitor cells, PF4 was assessed alone or in combination with IL-3 for effects on the total adherence and various kinds of adhesion molecules of KG1a cells as well as actin polymerization in KG1a cells. METHODS: The total adherence was assayed by crystal violet dye staining. The adhesion molecule expression was determined by FACS analysis. These adhesion molecule monoclonal antibodies individually blocked total adherence by MTT. F-actin content was monitored by fluorospectrophotometry. RESULTS: 100 ng/ml PF4 could increase the total adherence of KG1a cells by 80%. 20 ng/ml IL-3 could increase the total adherence of KG1a cells by 96%. When PF4 and IL-3 were combined, the total adherence could be promoted by 97%. Exposure of 1 x 10(6) cells/ml of KG1a cells to 100 ng/ml PF4 the increased total adherence of KG1a cells was mediated by PECAM-1 (CD31), CD44, LFA-1 (CD11a) and Mac-1 (CD11b) but not by P-selectin (CD62P) and E-selectin (CD62E). These adhesion molecule monoclonal antibodies could individually block total adherence for 34% 43%. Similar phenomenon was observed when IL-3 was added onto KG1a cells. Further study found that PF4 induced actin polymerization of KG1a cells. CONCLUSIONS: Our study indicated that PF4 promoted total adherence, as well as several adhesion molecule expression and actin polymerization of KG1a cells. The results suggest that PF4 may have therapeutic utility along with other cytokines by enhancing the total adhesion of hematopoietic stem/progenitor cells to promote the homing. PMID- 12905836 TI - [Effects of FPIL6/IL2 on the differentiation and proliferation of lymphoid in the ML-IC assay]. AB - OBJECTIVE: To develop an in vitro assay that allows the culture and identification of a single human bone marrow progenitor closely related to hematopoietic stem cell, which is more primitive than LTC-IC, and to find an efficient culture conditions for NK-IC expansion. METHODS: Fusion protein IL6/IL2 was reconstructed and expressed in E. coli DH5 alpha. ML-IC was determined by watching if the single cell can give rise to secondary progenitors with both LTC IC and NK-IC characteristics. LTC-IC frequency was determined by the CFC clonogenic methylcellulose assay. NK-IC frequency was determined by phenotyping CD56 positive NK cells. The effect of FPIL6/IL2 on the expansion of NK-IC was examined by comparing the colony number of NK cells before and after the culture. RESULTS: After the initial 4-week expansion culture, we showed that (25.75 +/- 5.68)% of freshly sorted Lin-/34+/DRdim cells were able to generate functional NK IC in one or more secondary FPIL6/IL2 cultures, whereas (6.81 +/- 1.97)% in the control. A total of 102 NK-IC cells were present when were cultured for 6-7 weeks in FPIL6/IL2 expansion medium, which was much higher than the 33 NK-IC cells in the control. CONCLUSION: ML-IC assay will prove useful to assess a very primitive hematopoietic cell with multilineage generative capacity. FPIL6/IL2 is capable of initiating and promoting NK-IC expansion greatly in ex vivo cultures in terms of net-conservation and net proliferation. PMID- 12905837 TI - [Effects of murine bone marrow endothelial cell conditioned medium on the growth of yolk sac hematopoietic stem cells and progenitors]. AB - OBJECTIVE: To investigate the effects of murine bone marrow endothelial cell conditioned medium (mBMEC-CM) on the growth of yolk sac hematopoietic progenitors. METHODS: The serum-free mBMEC-CM was obtained from subcultures of murine endothelial cell line derived from bone marrow which was established in our laboratory. The murine yolk sacs were harvested on day 8.5 postcoitus (pc) and incubated with 0.1% collagenase in 10% fetal calf serum at 37 degrees C for 40 minutes. Yolk sac cells were incubated in tissue culture dishes at 37 degrees C for 1 hour. Nonadherent cells were collected for semisolid culture assay of granulocyte-macrophage colony forming unit (CFU-GM) and high proliferative potential-colony forming cell (HPP-CFC) after being cultured in DMEM with 10% mBMEC-CM and 10% FBS for 24 hours. The number of CFU-GM and HPP-CFC was counted at day 7 and day 14 respectively. RESULTS: The growth of CFU-GM and HPP-CFC was supported by mBMEC-CM with GM-CSF. mBMEC-CM could induce the proliferation and differentiation of yolk sac hematopoietic stem cells and progenitors in liquid culture system. The percentages of CFU-GM and HPP-CFC compared with the 0 hour control were (119.5 +/- 5.7)% and (130.8 +/- 9.8)% respectively after 24 hours liquid culture (P < 0.05). The expansion effects of mBMEC-CM on CFU-GM and HPP CFC were enhanced by compounded with flt3 ligand (FL) and thrombopoietin (TPO). The percentages of CFU-GM and HPP-CFC compared with the 0 hour control were (132.0 +/- 6.2)% and (176.9 +/- 12.8)% respectively after 24 hours liquid culture (P < 0.01). CONCLUSION: Murine bone marrow endothelial cell conditioned medium could support the growth and proliferation of yolk sac hematopoitic stem cells and progenitors, and this promoting effect was further enhanced by addition of FL and TPO. PMID- 12905838 TI - [Osteogenic differentiation of murine yolk sac mesenchymal stem cells in vitro]. AB - OBJECTIVE: To investigate the potential of yolk sac mesenchymal stem cells in osteogenic differentiation. METHODS: Murine yolk sacs were harvested on day 8.5 postcoitus, yolk sac cells were obtained after the yolk sacs were digested by 0.1% type I collagenase for 1 hour, the non-adherent cells were removed after being cultured for 1 hour. The adherent cells were cultured in DMEM containing of 5 ng/ml bFGF and 15% FBS, and passaged when they became subconfluent. The morphologic characteristics, and AKP, BMP-2, as well as type I, III collagen of the yolk sac adherent cells were observed and tested. The attached cells were treated with 10(-8) mol/L dexamethasone, 10 mmol/L beta-glycerophosphate, and 50 micrograms/ml vitamin C at passage 4. Alternations of morphological characteristic, AKP activity, collagen of type I, III and mineralization were detected. RESULTS: Pure mesenchymal stem cells which were of spindle shape, uniform in size, positive in type I, III collagen staining and weak positive in AKP activity could be induced to pleomorphism osteoblast-like cells in vitro. The cells were transformed from spindle shape to polygonal cells which were positive in type I collagen, negative in type III collagen, strong positive in BMP-2, and positive in Von Kossa's stain at week 8. The polygonal cells could form nodular structure and their AKP activity was increased. All these were coincidence with the characters of osteoblast. CONCLUSION: Yolk sac mesenchymal stem cell can be purified and induced to osteoblast in vitro. PMID- 12905839 TI - [Identification and isolation of mesenchymal stem cells from human fetal pancreas]. AB - OBJECTIVE: To isolate and identify the phenotype and biological characteristics of pancreas derived mesenchymal stem cell. METHODS: Fresh pancreas of 4-5 months old aborted fetus was dissected free from connective tissue, and was cut into small pieces. The adherent cells were harvested and subcultured, after the third subculture, the cells were used for examination. Cell cycle was analyzed by measuring DNA content by FACScan flow cytometer. Phenotype of MSCs was analyzed by immunohistochemical SA technique and differentiated cells were identified by relevant specific staining. RESULTS: Fetal pancreas derived cells gave rise to a population of adherent cells characterized by the presence of a predominant cell type with a typical fibroblast like morphology. By transmission electron microcopy, MSC had few endoplasmic reticulums and mitochondrias. During the log phase of growth, MSC proliferated with a two fold population at 30 h. MSC can be ex vivo expanded by successive cycles of trypsinization, seeding, and culture. Under these conditions, MSC had capability of passaging up to 30 times without displaying significant changes in morphology, with 2-fold increase in cell number after each passage. This indicates the high ex vivo expansion potential of MSC. The results showed that the yield of CFU-Fs was above 200 clones even after the 6th passage. Cell cycle analysis by flow cytometry revealed that more than 83% of cells were in the G0/G1 phases, while a small population of cells were actively engaged in proliferation (S + G2 + M = 17%). We also showed that more than 86% of cells were positive stained by FITC labeled CD44, CD29, CD13, and only about 1% of cells were positive for CD34, HLA-DR. Expression of collagen I, III was positive while vWF was negative. In the differentiation study, we found culture expanded pancreas MSCs could be directed into the osteogenic lineage as detected by osteoblastic morphology, expression of alkaline phosphatase, modulation of osteocalcin mRNA production and the formation of a mineralized extracellular matrix. We also found that MSCs could give rise to the adipogenic and chondrogenic lineage as evidenced by accumulation of lipid-rich vacuoles within cells and the expression of lipoprotein lipase mRNA or the expression of collagen II and the deposition of proteoglycans. CONCLUSION: Mesenchymal stem cells existing in human pancreas can be isolated by their adherent ability and should be essential to sustain a steady supply of primitive cells in tissue remodeling. PMID- 12905840 TI - [Longitudinal study of the growth of craniofacial widths in 13-18 years adolescents with normal occlusion]. AB - OBJECTIVE: To study the growth characters of the craniofacial widths and then perusing its growth pattern. METHODS: 26 adolescents with normal occlusions were selected. A posteroanterior cephalogram was taken for every subject at the same time each year for 6 successive years. Computer-aided cephalometrics was carried out. RESULTS: (1) From age 13 to 18, most items of the craniofacial widths were bigger in male than in female. With the increase of age, the items with gender differences keep stable, except the width of L-Anter, L-Ore and L-Nas. (2) Compare with the growth completion of the craniofacial widths in 18 years old, they had completed more than 92% in 13 years old, and the L-Eur, L-Anter, L-Lo, L Max, L-UM and L-LM were almost complete as well. The growth completion quantity of L-Ore, L-Zyg, and L-Nas in female was larger than that in male. (3) The growth of L-Mas, L-Nas, and L-Zyg could keep on to the age of 18. The L-Max maitained stable in female after 13 years old, while there was a growth from age 13 to 17 in male. (4) From age 13 to 18, the tooth arch width kept stable, while the cusp of the lower canine tends to be converging. CONCLUSION: The present findings may aid orthodontists to make rational diagnosis and treatment planning. PMID- 12905841 TI - [Urinary endometriosis]. AB - OBJECTIVE: Aimed at illustrating the means of early diagnosis for prevention of the impairment of kidney function, the results of conservative medical therapy and the surgical treatment in patients with endometriosis of urinary tract. METHODS: Surgical interventions were done in 5 cases with pathological of endometriosis. Ureters were involved by endometriotic lesion in 3 patients, ureters and bladder in one and urethra in another patient. All of the cases were shown at least partial obstruction of the urinary tract by clinical symptoms, diagnostic examinations and the outcomes of various kinds of treatment. RESULTS: Hydropnephrosis and hydroureter were shown by ultrasonography, IVP or retrograde pyelography in one case of ureteral obstruction. A small lump of endometriotic foci along the urethra was detected by manual examination and ultra-sonography in one patient. The relieving of the symptoms and signs of the obstruction of the urinary tract in two cases by GnRH-a or progest in was the evidence of the endometriotic lesion. There were two patients underwent surgical resection of the endometriotic foci surrounding the ureter after medical therapy. CONCLUSIONS: Urinary tract endometriosis are rarely seen and usually escape the attention of the physicians, while they usually cause obstruction of the urinary tract and finally impairment of kidney function if not treated in time. This kind of disease can be easily diagnosed by imaging detection with ultra-sonography, IVP or retrograde pyelography. Conservative treatment with GnRH-a and long acting progesterone as well as surgical treatments are effective for this kind of disease. PMID- 12905842 TI - [Inhibitory effects of lovastatin on the proliferation and cell cycle phase of cultured human glomerular mesangial cells in vitro]. AB - OBJECTIVE: To study the effects and mechanism of lovastatin on cell cycle phase and proliferation of cultured human glomerular mesangial cells in vitro. METHODS: HMC proliferation was determined by 3H-Thymidine incorporation. HMC cell cycle was measured by flow cytometric analysis. RESULTS: Lovastatin was found to inhibit HMC proliferation in a dose-dependent manner. Flow cytometric analysis demonstrated that lovstatin induced G1/S transition arrest. Concomitant addition of mevalonate or farnesol restored all the inhibitory effect of lovstatin on HMC. CONCLUSION: Lovastatin is a HMC proliferation inhibitor. It provides an experimental evidence for re-evaluate renal protective effect of HRI, which has already been widely used in clinical treatment. PMID- 12905843 TI - [Construction of herpes simplex virus type I glycoprotein D DNA vaccine and its preliminary study]. AB - OBJECTIVE: The goal of this study was to construct a eukaryotic expression plasmid containing the gene encoding herpes simplex virus type I glycoprotein D (HSV-1, gD) and evaluate its utility for DNA immunization in mice. METHODS: The gD gene was amplified from viral DNA using PCR with EcoR I and BamH I restriction sites encoded on 5' and 3' ends, respectively. The PCR fragment was inserted into the transfer vector pGEM-T Easy. gD was then cut from this vector and inserted into the EcoR I and BamH I sites in the pcDNA3.1 at the multiple cloning sites (MCS). The recombinant plasmid, pcDNA3.1-gD1, was transfected into COS-7 cells using Lipofectamine according to the manufacture's instructions. The expression of the glycoprotein D was analyzed by immunoblotting of the cell lysates. 4-6 weeks old BALB/C mice were given two injections at tibia anterialis muscle, each containing 100 micrograms of plasmid DNA, on days 0 and 15. pcDNA3.1 was used as negative control. Blood samples were taken from all mice at weeks 0, 2, 4, and 6 after the first inoculation. Standard indirect ELISA was employed to evaluate the levels of specific total Ig in serum. RESULTS: The recombinant plasmid was confirmed with restriction digestion and sequencing to contain target gene segment and expressed in COS-7 cells in vitro shown by Western blotting. The pcDNA3.1-gD1 immunized group induced specific antibody response as compared to the negative control, and the titer was about 1:2000. CONCLUSIONS: The recombinant plasmid pcDNA3.1-gD1 is potential to be used as a candidate vaccine, for the treatment of HSV-1 infection. PMID- 12905844 TI - [The early pathological changes of KKAy mice with type 2 diabetes]. AB - OBJECTIVE: To investigate the early pathological changes of KKAy mouse model of type 2 diabetes. METHODS: Five male KKAy mice and six C57BL mice each were studied at the age of 8, 16, 20 and 24 weeks. With each mouse a 24-hour urine collection was made for the tests of proteinuria. Plasma insulin, lipids, serum creatinine and urea were also measured. Renal tissues were observed to assess glomerular and tubulointerstial pathology. RESULTS: The KKAy mice developed hyperglycemia, hyperinsulinemia and obesity by 16 weeks of age (P < 0.01). The proteinuria increased with the increasing of age (P < 0.005), but there were no changes in control. The glomerular hypertrophy was observed in KKAy mice at the age of 16 weeks. Computer map analysis system (CMIAS) indicated the expansion of mesangial matrix in KKAy mice with ageing. There was significant tubular dilation, accompanied with focal tubular atrophy and interstitial fibrosis. On electron microscopy, GBM undergo progressive thickening (P < 0.01), accompanied with podocytes fusion and increasing of proteinuria. CONCLUSIONS: KKAy mice developed hyperglycemia, hyperinsulinemia and obesity after 16 weeks, with proteinuria, mesangial matrix accumulation, GBM thickening and tubular dilation. It was considered a good animal model for the early pathology changes of DN. PMID- 12905845 TI - [Surgical treatment of spine metastatic tumor]. AB - OBJECTIVE: To explore the value of surgical treatment on spine metastatic tumor. METHODS: 31 cases of spine metastatic tumor with severe pain or/and neurological deficit were selected and treated with various operative methods. RESULTS: Except for 4 cases, all the patients survived from 3 months to 7 years (average 17.6 months). The patients had no complication during treatment. The symptoms and neurological functions recovered with various extent after operation. CONCLUSIONS: Surgical treatment should cautiously be adopted. When the spine metastatic tumor with severe pain and neurological deficit has no response to the conservative methods, the surgical treatment with decompression and internal fixation should be performed to improve the life quality for the patients. PMID- 12905846 TI - [Fat at sacroiliac joints: MR imaging findings (comparison and analysis of 18 volunteers and 52 patients with ankylosing spondylitis)]. AB - OBJECTIVE: In aid of MR imaging diagnosis of ankylosing spondylitis (AS), MRI findings of sacroiliac (SI) joints in volunteers were compared with those in patients with AS, with special reference to the fat findings. METHODS: 18 normal volunteers and 52 AS patients underwent the MRI examinations with the same protocols, i.e., SE T1WI, FSE T2WI, GE T2*WI sequences were taken using a 0.3T imager. All the images were observed and analyzed by two radiologists. RESULTS: Symmetric or patchy distribution fat component either in sacrum and/or ilium was detected in 52.8% of SI joints of volunteers. Thin zone distribution of fat was seen in 35.6% of SI joints of AS patients, which was adjacent to the erosion and sclerosis. In another 25.0% of SI joints of AS patients, the distribution of fat component was found similar to that of the normal volunteers. Cartilage changes were detected in 85.6% of the SI joints of AS patients. In addition, bone erosion, sclerosis and subchondral bone marrow edema were also detected. CONCLUSION: Subchondral fat infiltration associated with erosion and sclerosis should be considered as sacroiliitis. The knowledge of the characteristics of fat component could be helpful for evaluating sacroiliac joints. PMID- 12905847 TI - [Enzyme-amplified time-resolved fluorescence detection for nucleic acid hybridization assays]. AB - OBJECTIVE: To develop a new nonisotopic detection method of enzyme-amplified time resolved fluorescence (EATRF) or enzyme-amplified lanthanide luminescence (EALL) for nucleic acid hybridization assays, which can be applied extensively in clinical diagnosis. METHODS: The method combines the high affinity of biotin streptavidin system, amplification of enzyme, and inherent advantage of lanthanide chelate with the background elimination of time-resolved fluorescence detection. The conversion of 5-fluorosalicyl phosphate to 5-fluorosalicylic acid (5-FSA) by alkaline phosphatase. The salicylic acid product forms a luminescent ternary chelate with Tb3+ and EDTA. RESULTS: The dynamic range of the standard curve of EATRFA for nucleic acid hybridization assay was very wide, the range was more than third order of magnitude. The detection sensitivity was about 10 pg of target sequence. When the known target sequence was 20, 10 and 2 ng, the ratio of measured amount to known amount was 110%, 90% and 115% respectively. The main experimental conditions, for example, the irradiating time of ultraviolet rays, the concentrations of biotinylated probe, AP-SA, 5-FSAP and Tb-EDTA and the methods of washing in the related steps, have been optimized. A new stable technology of fluorescence has been developted. CONCLUSIONS: EATRF detection for nucleic acid hybridization assays is a new sensitive simple method, which has a great prospect. PMID- 12905848 TI - [A technique of continuous medical image digital restoration]. AB - OBJECTIVE: To set up a new technique of continuous medical image digital restoration for electronic record management. METHODS: According to Freeman's chain-code principle, 92 fetal cardiotocography were scanned, thinned, trailed, and chain-coded by computer system. RESULTS: 92 paper-recorded cardiotocography, were transformed to digital data and saved by computer, 23 parameters were produced, which could be used in further study. CONCLUSION: Capacity of a restoring instrument can be greatly saved since the using of digitalized restoration, so that more useful data can be stored in doctor's workstation. Digitalized data can be much easier used in electronic record management, for convenient studies research. PMID- 12905849 TI - [Effects of NBP on ATPase and anti-oxidant enzymes activities and lipid peroxidation in transient focal cerebral ischemic rats]. AB - OBJECTIVE: The aim of the present study was designed to explore the effect of (+/ ) -3-n-butylphthalide (NBP) on ATPase, anti-oxidant enzymes activities and lipid peroxidation of mitochondria and cerebral cortex in rats subjected to 24 hours of reperfusion following 2 hours of cerebral ischemia (tMCAO). METHODS: Activities of SOD (Superoxide Dismutase), GSH-Px (glutathione Peroxidase,) and CAT (Catalase), and MDA level of mitochondria or cortex were measured by using biochemical methods in tMCAO rats. RESULTS: (1) The activities of mitochondrial Na+K(+)-ATPase, Ca(2+)-ATPase and Mg2+ ATPase were found to decrease significantly in the vehicle group (ischemia + saline). Pre-treatment with NBP (5, 10, 20 mg/kg, i.p.) 10 min before tMCAO markedly enhanced the activities of Na+K(+)-ATPase and Ca(2+)-ATPase, compared with vehicle group. (2) The activities of SOD and mitochondrial GSH-Px were decreased and MDA level increased in vehicle groups as compared with that in sham group (non-ischemia + saline). NBP (20 mg/kg, i.p.) significantly enhanced total mitochondrial SOD activity, and also enhanced cerebral cortex total SOD activity (in 5, 10, 20 mg/kg groups). However, it had no obvious effect on CuZn-SOD activity. NBP (20 mg/kg i.p.) markedly increased mitochondrial (but not in cerebral cortex) GSH-Px activity; NBP 10, 20 mg/kg markedly decreased mitochondrial MDA level compared with that in vehicle group (P < 0.05). (3) The action of raceme NBP on the increase of the activities of ATPase and antioxidative enzymes seemed to be beneficial than that of (-) -NBP or (+) NBP. CONCLUSION: The results suggest that NBP improves energy pump and subsides oxidative injury which may contribute to its anti-neuronal apoptotic effect. PMID- 12905850 TI - [The effect of substance P on functional proteins in human neutrophil]. AB - OBJECTIVE: To explore the effect of substance P (SP) on the functional proteins on plasma membrane of neutrophil (Np). METHOD: The response of Np to SP was examined by measuring the level of respiratory burst, the activities of ACP and ALP, the fluoroscopy intensity of CR3, CD45 and FM-LP. RESULTS: It was found that SP could increase respiratory burst of Np, decrease the activity of acid phosphatase (ACP), but had no effect on alkaline phosphatase (ALP). SP could also promote the amount of CD45, complement receptor type 3 (CR3) and N-Formyl-Met-Leu Phe (FMLP) receptors. CONCLUSION: The results showed that the effects of SP on functional proteins in human Np membrane were universality and diversity. It implied that SP could affect various inflammation responses in Np. PMID- 12905851 TI - [Analysis of clinical manifestations of pharyngeal and laryngeal amyloidosis]. AB - OBJECTIVE: To study the clinical manifestations of pharyngeal and laryngeal amyloidosis. METHODS: Retrospective analysis of the clinical data of the inpatients with systemic and local amyloidosis. RESULTS: Of the 25 cases, sex ratio was 21:4, and average age was 48 years. Seventeen cases were systemic, of which 7 patients died in 2 years since diagnoses were made. One case was local bronchial amyloidosis. Seven cases were local primary pharyngeal and laryngeal amyloidosis and no affirmative death report was recorded since diagnoses were made. CONCLUSIONS: Amyloidosis occurs more likely in male and middle- and old aged patients. Pharyngeal and laryngeal local primary amyloidosis have relatively better prognosis than that of systemic amyloidosis. Follow-up is important because local recurrence may be happened. PMID- 12905852 TI - [Progress in the studies on the relationship between the polymorphism of P22phox gene and coronary heart disease]. AB - Reactive oxygen species (ROS) are thought to contribute to the progression of cardiovascular disease (CVD). At present, it has been known that the NADH/NADPH oxidase system is the major source of superoxide in the vascular system. Cytochrome b-245 (P22phox), which is a critical component of NADH/NADPH oxidase, plays an important role in electron transport and producing the superoxide anion. It is considerable to take attention to whether the polymorphism of P22phox gene is associated with a risk of coronary heart disease (CHD). To distinguish the relationship between them will be beneficial to elucidate the genetic mechanisms of CHD, and develop a new genetic marker to provide theoretical base for the prevention and cure of CHD. PMID- 12905853 TI - [Growth inhibition of human nasopharyngeal carcinoma cell CNE-2L2 caused by suppression of 6A8 alpha-mannosidase expression]. AB - OBJECTIVE: To study the relationship between 6A8 alpha-mannosidase expression and growth of CNE-2L2 cells, a human nasopharyngeal carcinoma cell line. METHODS: Recombinant adeno-associated virus vector (rAAV) was used as a mediator to transfer an antisense or a sense fragment of 6A8 cDNA into CNE-2L2 cells. 6A8 alpha-mannosidase expression was detected by means of mAb 6A8a staining, alpha mannosidase activity assay and ConA binding test. Cell growth was examined by means of MTT and colony formation. Tumor growth at the inoculated site of the cells in nude mice was detected after 8 weeks. RESULTS: Transduction of antisense 6A8 could reduce the expression of 6A8 alpha-mannosidase. In comparison to those in controls, the wild type, the mock-transduced and the sense 6A8-transduced cells, the MTT value, the colony number formed and the tumor weight grown at the inoculated site of cells were significantly decreased in the antisense 6A8 transduced cells (P < 0.001). CONCLUSION: Decreased expression of 6A8 alpha mannosidase caused an inhibition of CNE-2L2 cell growth. PMID- 12905854 TI - [6A8 alpha-mannosidase catalyzes p-nitrophenyl-alpha-D-mannopyranoside]. AB - OBJECTIVE: To confirm the alpha-mannosidase nature of the protein encoded by 6A8 cDNA. METHODS: 1) To construct a full-length 6A8 cDNA based on the three cloned DNA fragments by means of gene recombinant technique; 2) To insert the 6A8 cDNA into eukaryotic expression vector pCDI; 3) To transfect the recombinant pCDI-6A8 into COS-7 cells; 4) To characterize the nature of the protein encoded by 6A8 cDNA by means of enzymic activity assay and Western blotting assay. RESULTS: The constructed 6A8 cDNA was the right cDNA in sequence. The enzymetic activity of the homogenate of COS-7 cells transfected with pCDI-6A8 was 3-4 times higher than that of the cells transfected with the mock or the wild cells. The enzymetic reaction could not be inhibited by swainsonine. Western blot showed a band of 120,000 recognized by mAb 6A8. The band in the cells transfected with pCDI-6A8 cDNA was much darker than that in the cells transfected with the mock or in the wild cells. CONCLUSION: The protein encoded by 6A8 cDNA is a kind of alpha mannosidase, which belongs to type II alpha-mannosidase. PMID- 12905855 TI - [The effect of Beijing Fangshan Cardiovascular Prevention Program on the incidence and mortality of stroke]. AB - OBJECTIVE: To explore potential effective measures for lowering incidence and mortality of stroke in rural community population of China. METHODS: Beijing Fangshan Cardiovascular Prevention Program (BFCP), under whole population and high risk individuals strategies with measures of health education and hypertension control, were launched in 1991 in five communities including intervention communities (IC), about 66,000 residents, and control communities (CC), about 54,000 residents, in Fangshan, Beijing suburb. RESULT: Incidences of stroke averaged 235.23 per 100,000 and 289.22 per 100,000, for IC and CC respectively, with a statistically significant difference, and mortalities of stroke averaged 80.63 per 100,000 and 98.01 per 100,000, for IC and CC respectively, with a statistically significant difference, during years of 1992 to 1999. The net change of stroke incidence was 126.13 per 100,000 in IC versus CC, with a statistically significant difference. Incidences of stroke increased by 11.63% and 75.27%, for IC and CC respectively, while mortalities of stroke decreased by 46.80% and 22.82%, respectively, for IC and CC from years of 1992 to 1999. CONCLUSION: BFCP has yielded obvious effect on controlling incidence and mortality of stroke in rural community population, but the trend of stroke incidence increasing was still not restrained radically. PMID- 12905857 TI - [Aneurysmal model induced by elastase]. AB - OBJECTIVE: To induce the aneurysms by corrosion of aneurysmal wall with elastase, imitating clinical intracranial aneurysms patho-morhologically. METHODS: The elastase-corroded arterial segments were transplanted onto lateral walls of carotid arteries. Follow-up and pathological examination of the aneurysms were made at two weeks and one month after construction. RESULTS: The aneurysms formed by the transplantation of elastase-corroded arterial segments were found persistent or enlarge a little within one month. The aneurysms were purplish red and easy to rupture when exposed. The walls were deprived of elastic layer, and constructed by thin connective tissue. CONCLUSIONS: The aneurysms made by transplanting elastase-corroded arterial segments are simple to built, while morphological and pathological characteristics are demonstrated good clinical mimicry, we believe our aneurysm model may well substitute the most popularly used aneurysms made of venous pouches. PMID- 12905856 TI - [Clinical value of induced sputum test in monitoring airway inflammation in asthma]. AB - OBJECTIVE: To explore the clinical value of induced sputum test in monitoring airway inflammation in asthmatic patients. METHODS: The induced sputum test was done in 12 normal subjects and 21 asthmatic subjects; The levels of interleukin-8 (IL-8), eosinophil cation protein (ECP), malondialdehyde (MDA) in plasma and induced sputum in all subjects were measured. RESULTS: The levels of IL-8, ECP, MDA in plasma and induced sputum in unstable asthmatic patients were significantly higher than those in stable asthmatic (P < 0.01), in which the levels of IL-8, ECP, MDA in both plasma and induced sputum were significantly higher than those in normal subjects(P < 0.05); In all asthmatic patients, the levels of IL-8, ECP, MDA in induced sputum were significantly higher than those in plasma. CONCLUSION: The levels of IL-8, ECP, MDA in induced sputum could be used for monitoring airway inflammation in asthmatic patients. PMID- 12905858 TI - [Alzheimer-like protein phosphatase deficiency leads to abnormal phosphorylation and accumulation of neurofilaments]. AB - OBJECTIVE: To explore the effect of Alzheimer-like protein phosphatase deficiency on neurofilament phosphorylation. METHODS: Cell culture, light microscopy, immunocytochemistry and biochemistry techniques were used to make a phosphatase deficient cell model, to detect cell morphology, neurofilament phosphorylation and distribution, cell viability and activity. RESULTS: Non-phosphorylated neurofilament recognized by SMI32 was detected both in cell body and cell processes, it was extremely enriched in cell bodies; Phosphorylated neurofilament bound to SMI34 was mainly determined in cell processes and cell surface. After treatment with okadaic acid (OA), non-phosphorylation-dependent antibodies SMI32 staining was significantly decreased in the cell body, whereas phosphorylated neurofilament reacted with SMI34 was strikingly increased in immunocytochemistry and Western blot, and prominently accumulated to the same cell location. Accompanied with hyper-phosphorylation and accumulation of neurofilament, dose dependent cell toxicity was observed by okadaic acid treatment. CONCLUSION: Deficiency in protein phosphatase induces in neuroblastoma cell line, neurofilament phosphorylation and accumulation, which is involved in Alzheimer neurofibrillary degeneration. PMID- 12905859 TI - [Comparative study of the expression and phosphorylation of neurofilament proteins of brain gray matter in Alzheimer's disease]. AB - OBJECTIVE: To explore the role of abnormal modification of neurofilament proteins (NFPs) in the pathogenesis of Alzheimer disease (AD), and the characteristics of NFPs in distribution and solubility. METHOD: Ultra-centrifugation and Western blot were utilized to isolate and analyze NF subunits. Ultra-centrifugation and Western blot were utilized to observe the effect of abnormal modification of neurofilament proteins (NFPs) on the characteristics of NFPs in distribution and solubility in the pathogenesis of Alzheimer disease. RESULTS: In human brain gray matter, NF subunits were mainly detected in particular fraction. Compared with Huntington's disease (HD), the content of NF subunits was increased in AD brain gray matter and the increased level of NF-H and NF-M were mainly in abnormal hyper-phosphorylate form. CONCLUSION: Abnormal phosphorylation and accumulation of NFPs are involved in pathology of AD brain. PMID- 12905860 TI - [An association analysis of apolipoprotein E genotypes with Alzheimer's disease in Chinese population]. AB - OBJECTIVE: To analyse the association of apolipoprotein E genotypes with Alzheimer's disease in Chinese population. METHODS: Using PCR and restriction enzyme digestion, we have analyzed ApoE allele frequency of Alzheimer's disease (AD) patients and healthy controls. RESULTS: Among the 56 cases of AD, the frequency of ApoE epsilon 2, ApoE epsilon 3, ApoE epsilon 4 are 3 (2.7%), 84 (75.0%), 25 (22.3%) respectively, while in the 67 controls they are 9 (6.7%), 115 (85.8%), 10 (7.5%) respectively. The frequencies of the two groups have significant difference (P < 0.01) and the frequency of ApoE epsilon 4 allele is higher in AD than that in the health controls and the statistical treatment suggest that there is significant difference (chi 2 = 10.99, P < 0.01, OR = 3.59, 95% CI = 1.54-8.41). The frequency of the ApoE epsilon 3 is lower in AD than that in the health controls and the difference is also statistically significant(P < 0.05). CONCLUSIONS: Our results are consistent with previous work in that proving the ApoE epsilon 4 allele is one of the risk factors of AD. The results also provide a support for the protection effect of ApoE epsilon 3 allele in developing AD. PMID- 12905861 TI - [A retrospective analysis of 8 cases with primary central nervous system lymphoma]. AB - OBJECTIVE: To make an inquiry into clinical features, treatment, and prognosis of primary central nervous system lymphoma (PCNSL). METHOD: A retrospective analysis was carried out in a group of 8 patients confirmed diagnostic PCNSL by clinical findings and pathological examination. RESULTS: The common clinical manifestation of PCNSL consisted of increased intracranial pressure, seizures, focal neurologic deficits. Studies of cerebrospinal fluid revealed protein elevation in the most patients. Neuroradiological examinations with CT and MRI both showed solitary or multiple lesion located cerebral hemisphere, periventricular, basal ganglia, corpus callosum. Pathological examination showed B-cell malignant lymphoma. CONCLUSION: The patients with PCNSL usually did not show characteristic clinical manifestations or findings in imaging scanning. The brain biopsy pathological examination might be the only reliable method to confirm the diagnosis of the disease. PMID- 12905862 TI - [Identification and cloning of a novel full ORF gene of P. falciparum PDZ domain containing protein]. AB - OBJECTIVE: To identify and clone a novel full ORF gene of P. falciparum PDZ domain containing protein (PfPCP) and study its expression at the erythrocytic stage. METHODS: The primers were designed according to the sequences from P. falciparum genomic database. The full ORF gene of PfPCP was amplified from mRNA of the erythrocytic stage of P. falciparum Hainan strain by RT-PCR. Bio informatics software were used to analyze PfPCP; PfPCP expression at the erythrocytic stage was studied with Western Blotting. RESULTS: The full ORF gene of PfPCP was 2,076 bp and encoded 691 amino acids. PfPCP contained the typical PDZ domain. Western blotting showed that PfPCP was expressed at the schizont sub stage of P. falciparum erythrocytic stage. CONCLUSIONS: The novel full ORF gene of PfPCP is cloned. As a PDZ domain containing protein, PfPCP is expressed at the schizont sub-stage of P. falciparum. PMID- 12905863 TI - [Determination of the binding site of testis-specific nucleoporin BS-63 to transportin (karopherin beta 2) and the proof of their combination in vitro]. AB - OBJECTIVE: To locate the binding site of testis-specific nucleoporin BS-63 to transportin (karopherin beta 2) and confirm their combination in vitro. METHODS: Constructed different fragments of C terminal BS-63 was employed to localize the binding site of the testis-specific nucleoporin BS-63 to transportin by yeast two hybrid system technique pull-down test was used to identify the interaction between the purified expressed fragments of BS-63 0.6 K and transportin in vitro. RESULTS: BS-63 binding site to transportin was shortened from 1.6 kb to 0.6 kb which included a Ran binding domain (RanBD). SDS-PAGE and Western blot tests confirmed the recombinant purified protein coded by 0.6 kb fragment of BS-63 cDNA could interact with transportin in vitro. CONCLUSIONS: In germ cells, the function of the testis-specific nucleoporin BS-63 localized at cytoplasmic side of NPC importing cargoes into nuclear may be accomplished by transportin cooperation. PMID- 12905864 TI - [The cellular mechanism of angiotensin II inhibition on BKCa of ECV304 cell and the protective effect of gingko leaf extract]. AB - OBJECTIVE: To observe the cellular mechanism of A II inhibition on Maxi conductance calcium activated potassium channel (BKCa) in ECV304 cell membrane. METHOD: Using the cell-attached configuration of patch clamp technique. RESULTS: A II receptor antagonist saralasin (10(-7) mol/L) may block the inhibitory effect of A II (10(-7) mol/L). Phorbol ester (5 x 10(-8) mol/L) potentiated the effect of A II, while NO (10(-10) mol/L SNP) decreased the effect of A II and gingko leaf extract (800 micrograms/ml) activated BKCa and opposed the effects of A II. CONCLUSION: A II receptor mediates the inhibitory effect of A II on BKCa in ECV304, and PKC is involved in this inhibition. NO and ginkgo leaf extract protect BKCa from the inhibition of A II. PMID- 12905865 TI - [Patient-controlled analgesia with lornoxicam in patients undergoing gynecological surgery]. AB - OBJECTIVE: To assess the analgesic effect and side effects of PCA with lornoxicam compared with morphine and tramadol. METHODS: 89 patients, scheduled for elective hysterectomy or hysteromyomectomy, were randomly divided into Group L, Group M and Group T. Three drugs administered i.v. via a patient-controlled analgesia for up to 24 h postoperatively. RESULTS: Efficacy was assessed by comparing total pain relief (TOTPAR) and sum of pain intensity difference (SPID) over 24 h. Statistically significant equivalence of lornoxicam, morphine and tramadol was shown by TOTPAR values 15.2 +/- 3.9, 16.4 +/- 3.5 and 15.9 +/- 4.4, by SPID values 10.3 +/- 3.1, 9.0 +/- 2.0 and 9.2 +/- 4.7, respectively (P > 0.05). Lornoxicam caused fewer adverse events than morphine and tramadol (10.0%, 26.7% and 17.2% of patients, respectively). CONCLUSION: The study suggests that lornoxicam provides an alternative to morphine or tramadaol for the treatment of postoperative pain. PMID- 12905867 TI - [An approach for proper recording of the unit discharge in auditory nerve and inferior colliculus]. AB - OBJECTIVE: To explore a proper recording of the unit discharge in auditory nerve and inferior colliculus for the observation of characteristics of response potential. METHODS: Single unit discharge in auditory nerve and inferior colliculus of healthy adult cat to acoustic stimulation and intracochlear electrical stimulation were recorded. With glass microelectrode. RESULTS: (1) the spontaneous discharge in auditory nerve primary fibers were identified. (2) The waveform of the spontaneous discharge in auditory nerve primary fibers was biphasic potential with positive pulse width 0.5 ms. (3) The discharge rate of auditory nerve ranged from several PPS to 100PPS. (4) The response in inferior colliculus to high frequency pure tone stimulation lacked phase locking. (5) The response in inferior colliculus to low frequency electrical stimulation in cochlea showed phase locking. CONCLUSIONS: It was successful to record the spontaneous discharges and auditory evoked responses in auditory nerve as well as the pure tone evoked response and electrically evoked response in inferior colliculus. However, the existing technical problem in recording the electrically evoked response in auditory nerve was to keep the response potential stable. PMID- 12905866 TI - [Comparative effects of carvedilol and metoprolol in preventing from left ventricular remodeling after acute myocardial infarction in rats]. AB - OBJECTIVE: To compare the effects of carvedilol and metoprolol in preventing from left ventricular remodeling (LVRM) after acute myocardial infarction (AMI) in rats. METHODS: Twenty-four hours after ligating left coronary artery, 105 surviving female SD rats were randomly assigned to AMI control, carvedilol 1 mg/(kg.d) and metoprolol 2 mg/(kg.d) groups. Sham-operated rats (n = 16) were selected randomly as non-infarction control. After four weeks of drugs therapy, hemodynamic studies and pathologic analysis were performed. Exclusive of MI size < 35% or > 55%, complete experimental variables were obtained in 46 rats, which were comprised of AMI (n = 11), carvedilol (n = 12), metoprolol (n = 11), and Sham-operated (n = 12) groups. RESULTS: Compared with sham-operated group, left ventricular (LV) end diastolic pressure (LVEDP), volume (LVV) and weight (LVW), were all significantly increased (P < 0.05-0.001), while maximal rate of rise and fall (+/- dp/dt) of LV pressure as well as their corrected values (+/- dp/dt/LVSP) were all significantly decreased (P < 0.01-0.001) in AMI group. In comparison with AMI group, the LVEDP and LVV were all significantly decreased (all P < 0.001), while +/- dp/dt and +/- dp/dt/LVSP were significantly increased (P < 0.05-0.001) in both carvedilol and metoprolol group, with LVW and RVW only decreased in carvedilol group (P < 0.05-0.01). CONCLUSIONS: 1. Carvedilol can effectively attenuate LVRM, and improve hemodynamics and LV function after AMI in rats, 2. Metoprolol has equivalent beneficial effects as carvedilol on hemodynamics, LV dilatation and function, but not LV hypertrophy. PMID- 12905868 TI - [Effects of 17 beta-estradiol on the release of nitric oxide from HUVEC]. AB - OBJECTIVE: The protective mechanism of 17 beta-estradiol on blood vessels was investigated to provide evidences for clinical use of hormone replacement therapy in postmenopausal women. METHODS: To observe the effects of 17 beta-estradiol (17 beta-E2), testosterone (T), homocysteine (Hcy), 17 beta-E2 combined with T and 17 beta-E2 combined with Hcy on the production of nitric oxide (NO) released from cultured human umbilical endothelial cells (HUVEC) in vitro. RESULTS: Preincubation with 10(-9) mol/L 17 beta-E2 8-48 hours significantly enhanced the release of nitric oxide from HUVEC versus the control, but 10(-9) mol/L T had no effects within 48 hours. Pretreatment with 10(-9)-10(-7) mol/L testosterone for 24 hours had no effects on the release of NO from HUVEC versus the control, but 10(-6), 10(-5) mol/L T significantly reduced the release of NO from HUVEC. HUVEC were treated by 10(-9)-10(-5) mol/L 17 beta-E2 and 10(-9)-10(-5) mol/L T respectively for 24 hours. 10(-9), 10(-8) mol/L T had no effects on the beneficial effect of 17 beta-E2 with regard to the production of NO from HUVEC, while 10(-5) mol/L T greatly attenuated the beneficial effect of 17 beta-E2 with regard to the production of NO from HUVEC. Pretreatment with 10(-4)-10(-3) mol/L Hcy for 24 hours greatly inhibited the release of NO from HUVEC. If treated HUVEC with various concentrations of 17 beta-E2 (10(-9), 10(-7), 10(-5) mol/L) and Hcy for 24 hours simultaneously, 10(-9)-10(-7) mol/L 17 beta-E2 can partly eliminate the effects of Hcy on the released of NO from HUVEC. CONCLUSIONS: T can not enhance the release of NO from HUVECs. The effect of 17 beta-E2 combined with T on the release of NO depends on the concentration of both 17 beta-E2 and T. When 17 beta-E2 and T are in low dose, HUVEC can release more NO. Hcy can inhibit the release of NO from HUVEC, concomitant treatment with a low dose of 17 beta-E2 may eliminate the effects of Hcy on HUVEC. PMID- 12905869 TI - [Down regulatory effects of platelet factor four (PF4) on total adherence and respiratory burst of human neutrophil]. AB - OBJECTIVE: The effect of platelet factor four (PF4) on human neutrophil function was studied. METHODS: Crystal violet dye staining, immunofluorescence labeling, PKC kit assay, NBT and HVA fluoro-spectrophotometry were applied to study the effect of PF4 on total adherence, integrin CD11b level, PKC level and respiratory burst level of resting human neutrophils and FMLP/or PMA-stimulated human neutrophils. RESULTS: It was found that PF4 slightly increased the total adherence of resting human neutrophils. There was no change on integrin CD11b level and respiratory burst level of resting human neutrophils after interaction with PF4. However, PF4 significantly down-regulated the total adherence, integrin CD11b level and respiratory burst level of human neutrophils stimulated by FMLP/or PMA. In addition, PF4 did not influence PKC level on resting and activated human neutrophil. CONCLUSION: These results indicate that PF4 plays a down regulation of the function of human neutrophils. Differing from other members of classical CXC-chemokine family, the signaling of PF4 is not through PLC-PKC signal pathway. PMID- 12905870 TI - [Impact of AFP promoter on the expressions of GFP in tumor cells]. AB - OBJECTIVE: To study the impact of AFP 5'flanking promoter (enhancer) on the expression of GFP in hepatocarcinoma cell. METHODS: Green Fluorescent Protein (GFP) reporter gene expression plasmid pcDNA3-GFP-AFP-w under the direction of AFP 5' flanking promoter (enhancer) was constructed by recombinant DNA technology and confirmed by restriction analyses. pcDNA3-GFP-AFP-w, pcDNA3-GFP and pcDNA3 were transfected into Hela and Bel7402 cells by lipofectin and selected by G418 respectively, after amplification of the positive cell clones, expression of GFP was detected by Western blotting and quantitatively analysed by GEL Doc 2000 digital image systems. RESULTS: The expression of GFP was lower in Bel-GFP-AFP-w than in Bel-GFP but was significantly higher than in Hela-GFP-AFP-w. CONCLUSION: GFP reporter gene plasmid pcDNA3-GFP-AFP-w under the direction of the 3.1 kb AFP 5'flanking promoter (enhancer) can be expressed in HCC Bel7402 cell definitely and specifically. PMID- 12905871 TI - [Relationship between alpha-ENaC gene Thr663Ala polymorphism and ischemic stroke]. AB - OBJECTIVE: To determine the relationship between Thr663Ala polymorphism in amiloride-sensitive epithelial sodium channel alpha-subunit (alpha-ENaC) gene and ischemic stroke in Chinese population. METHODS: Two hundred and ninety-four patients with cerebral infarction and 280 controls without stroke were screened by polymerase chain reaction (PCR) and corresponding Thr663Ala polymorphism probe hybridization. RESULTS: Thr663Ala polymorphism was in agreement with Hardy Weinberg equilibrium. Both the carriers proportion (70.75%) and the frequency (48.64%) of 663Ala allele in patients were more prevalent than that in controls (62.86% and 40.89%) (P < 0.05). The presence of at least one 663Ala allele conferred an odds ratio for stroke of 1.429 (95%CI: 1.009-2.025), and the proportion of attributable risk of population (PAR%) was 11.63%. CONCLUSION: Thr 663Ala polymorphism of alpha-ENaC gene may be a genetic risk of ischemic stroke in Chinese population. PMID- 12905872 TI - [Study of normal range of speed of sound in tibia in Beijing area]. AB - OBJECTIVE: To study the normal reference value of adult tibial speed of sound (tibia SOS) in Beijing region. METHODS: 951 normal volunteers (male/female = 461/490) were included in this study. Speed of sound were measured in the middle of tibia by ultrasound scan (Sound Scan 2000, Myrid, Isereal). According to their gender and age, all subjects were divided into different subgroup by 10 year-age period. RESULTS: The peak value of SOS presented in 30-39 years period in both male and female. The peak value was higher in males than in females. After reach the peak value, the tibial SOS begin to decline. The decreasing rate was fast in SOS were found at 50-59 period in female and over 50-59 years old in male. The tibia values of SOS of postmenopausal was obviously lower than that of premenopausal women. The relationship of menopausal duration and the values if tibial SOS was found to be a non-linear curve, which included 3 stages: the quick decrease, stabilization and further decrease. CONCLUSION: The tibia values of SOS provides a kind of meaningful diagnostic criteria of osteoporosis. PMID- 12905873 TI - [Analysis of 128 Wilson's disease]. AB - OBJECTIVE: To investigate the clinical feature and misdiagnosis of 128 patients with Wilson's disease and propose the points of diagnosis for this disease. METHOD: 128 patients with Wilson's disease from Jan. 1983-Apr. 2000 admitted to our hospital were reviewed. Their clinical feature and diagnosis condition, such as the early onset symptoms, misdiagnosis and hypersplenism were studied. RESULTS: 1. The first onset symptoms: 64 were liver dysfunction and 52 were manifestations of CNS (P > 0.05). 2. The average age of patients with liver dysfunction was significantly younger (13 +/- 3.5 years) than that of CNS manifestations (23 +/- 8.3 years, P < 0.05). 3. The positive rate of cornea Kayser-Fleischer ring was 96.8%; the value of copper oxidase absorbency was 0.051 +/- 0.024 (0.01-0.16) and its positive rate was 93%. 4. The overall misdiagnosis rate was 42.9% (55/128) and the predominant mis-diagnostic diseases were hepatitis A and chronic hepatitis. CONCLUSION: The clinical manifestations are diverse, which make the misdiagnosis rate very high. The age of liver dysfunction patients is significantly younger than that of the CNS manifestations patients. Cornea Kayser-Fleischer ring and copper oxidase absorbency are of high positive rate, as it can be carried out conveniently in clinics we recommends to use this diagnostic measure for Wilson's disease. PMID- 12905874 TI - [Compound EPSPs and action potential of mauthner cell evoked by skin stimulation in crucian carp]. AB - OBJECTIVE: To investigate the effects of afferent excitatory inputs from skin on the excitability of Mauthner cell (M cell) of crucian carp. METHODS: Multiple site intracellular recordings on the soma and the ventral dendrite (VD) of M cell and direct current stimulation on the skin surface of the fish's trunk were employed. RESULTS: Direct stimulation of the skin evoked 3 groups of compound excitatory postsynaptic potentials (EPSPs) on soma and VD of M cell. Group a EPSP had the lowest amplitude (< or = 0.35 mV), which was insensitive to high frequency stimulation. It had the shortest latency (mean 1.9 ms) and could be recorded on the soma and the proximal end of VD. Group b EPSP had the highest amplitude (< or = 9.7 mV), which was increased when the recording electrode was moved from the soma to the distal end of VD. The latency of Group b (mean 4.5 ms) was shorter than that of Group c, but longer than that of Group a. We firstly found that action potential could be induced on the basis of Group b in M cell by skin stimulation. Group c EPSP was our new finding, which could only be evoked by higher intensity (noxious) stimulus (> or = 100 V). It had the longest latency (mean 13.5 ms) with amplitude between those of a and b, and was very sensitive to high frequency stimulation. All the three groups of EPSPs were superimposed with spike-like transient potentials, which were believed to be the sign of electrical synapse activities. CONCLUSIONS: (1) Action potential can be induced on M cell by skin stimulation, which is in controversy with previous reports; (2) the noxious skin stimulation can induce a late EPSP (Group c) in M cell; (3) the neural pathway projecting from skin to M cell is composed of a set of neuronal chains with different number of synaptic relays, in which short chains projected concentrically to the soma and the proximal part of VD, while long chains projected mainly to the distal part of VD; (4) both electrical and chemical synapses may exist in short and long pathways. PMID- 12905875 TI - [The role of immune enhanced enteral nutrition on plasma amino acid, gut permeability and clinical outcome (a randomized, double blind, controlled, multi center clinical trail with 120 cases)]. AB - OBJECTIVE: To evaluate the role of arginine, RNA and omega 3 fatty acid enriched enteral nutrition. METHODS: The study was designed as a prospective, randomized, double blind, multi-central trial. It was an isocaloric and isonitrogenous intake in both groups. The protocol was approved by the Ethic Committee and, written informed consents were obtained. RESULTS: There were 120 patients enrolled in this protocol. After data were input to computer, open the code. 118 out of 120 patients completed the study and, 2 of them were dropped out. One is because the nasal jejunum tubes dropped and not willing to be replaced. Second patient had fistula of anastomosis on 4th days after operation. There were finally 60 patients in the study group and 58 in the control group. There were no liver or renal functions damage and, obvious adverse in both groups. Plasma amino acid profile: There was significant difference (delta) of plasma arginine levels pre- and after study [(33.7 +/- 58.5) mumol/L vs (-2.4 +/- 30.7) mumol/L] (P = 0.004). Intestinal Permeability (lactulose/mannitol ratio): The differences (delta) of lactulose/mannitol ratio pre- and after the study were 0.017 +/- 0.012 in study group and, 0.027 +/- 0.016 in control group. (P = 0.047). Immunological markers: Humoral immunity: The differences of IgM levels pre- and after the study were (0.6 +/- 0.4) g/L in study group and, (0.2 +/- 0.4) g/L in control group(P = 0.006). Cellular immunity: The differences (delta) of CD3 levels pre- and after the study were (3.8 +/- 5.2)% in study group and (0.3 +/- 6.5)% in control group (P = 0.01). In CD4, (3.4 +/- 5.3)% in study group and, (-0.3 +/- 5.7)% in control group (P = 0.032). Clinical Outcomes: There was no infection-related in study group and, 2 abdominal infection patients in control group. No significant difference was found between groups (P = 0.46). The hospital stays were (13 +/- 2.5) days in study group and, (14.5 +/- 3.0) days in control group (P = 0.004). The cost for full hospitalization was (15,122 +/- 6,279) Yuan in study group and, (17,403 +/- 7,091) Yuan in control group. There was 2,281 Yuan lower in study group (P = 0.07). The costs for nutritional drugs were (1,383 +/- 242) Yuan in study group and, (707 +/- 111) Yuan in control group. The difference was 676 Yuan higher in study (P = 0.001). CONCLUSION: Immune enhanced enteral nutrition had better plasma arginine level, intestinal permeability marker, IgM, CD3 and CD4. Also had less hospital stay and, less totaled hospital cost in study group. PMID- 12905876 TI - [Gene clone of autoantigen hnRNP A2/B1 and research on expression in synovium]. AB - OBJECTIVE: To investigate hnRNPA2/B1's role in rheumatoid synovitis by obtaining its cDNA sequence and analyzing its expression in synovium. METHODS: Total RNA was isolated from peripheral blood mononuclear cells (PBMC) and the RT-PCR was performed. The fragment was cloned into PUC-T1 plasmids and further sequenced. We detected it's expression in synovium with monoclonal antibodies and specific cDNA probes by immunohistochemistry and in situ hybridization. RESULTS: The fragment was identified by DNA sequencing. The expression in rheumatoid synovium was higher than that from osteoarthritis and the normal controls (P < 0.05). CONCLUSION: hnRNP A2/B1 cDNA has been cloned successfully and hnRNP A2/B1 might play an important role in rheumatoid synovitis. PMID- 12905877 TI - [Effects of recombinant human macrophage colony stimulating factor (rhM-CSF) on stromal cell derived during the time of mice colony formation unit-spleen (CFU-S) and its role on CD34+ cells expansion in vitro]. AB - OBJECTIVE: To investigate the effects of stromal cell derived from mice spleen during in vitro expansion of CD34+ hematopoietic stem cells from umbilical cord blood. METHODS: Irradiated and non-irradiated mice at day 10, 12 and 14 were sacrificed and their spleens were taken for collecting spleen cells. The cells were stimulated with recombinant human macrophage colony stimulating factor(rhM CSF) for collecting adherent spleen derived stroma cells. The effects of spleen derived stromal cells which treated with mitomycin served as a feeder layer, combined with different cytokines on in vitro expansion of purified cord blood CD34+ were assayed. Clonogenic assay and flow cytometry were employed to analyze phenotype of expanded cells and their ability to form different colony in vitro. RESULTS: 1) rhM-CSF efficiently stimulated spleen derived stromal cells to proliferate; 2) The mice stromal cells obtained during the time of colony formation-unit spleen (CFU-S) promoted the CD34+ cells to form colony in semisolid medium and, combined with cytokines, thereby efficiently expanded the hematopoietic cells from cord blood. After two weeks incubation, CD34 positive cells were 100 times more than original cells cultured and expanded cells still have the ability to form multi-lineage colony in vitro. CONCLUSION: rhM-CSF can stimulate the stromal cells derived from CFU-S to proliferate and support the survival and expansion of CD34+ cells separated from cord blood. PMID- 12905878 TI - [Gene chip technology and its advances in medical science]. AB - Gene (DNA) chip or DNA microarray is a new technique developed during human genomic research programs. When DNA chip containing thousands of DNA or oligonucleotide are hybridized to labeled samples, gene expression, DNA sequencing as well as DNA mutation and polymorphism can be analyzed with high efficiency in large scale. We are able to perform this technique on dozens of specimens at once in tumor differentiation, tumor typing, tumor diagnosis, tumorigenesis, and new tumor-associated genes discovery. PMID- 12905879 TI - [Survivin--a new tumor-specific anti-apoptosis factor]. AB - Survivin (SVV) is a new member of IAP-family (inhibitor of apoptosis, IAP) proteins. It can inhibit apoptosis induced by a variety of factors, and plays important roles during mitosis and cytokinesis of cells. Studies indicated that it is a tumor-specific anti-apoptosis factor. Survivin is expressed during embryonal development but lacks expression in terminally differentiated adult tissues. Interestingly, it becomes reexpressed in transformed cell lines and in a variety of human tumors. Its overexpression is prognostic of tumors and is implicated in the resistance to chemotherapy of tumor cells. SVV may be a new and widely applicable target for gene- and protein-based therapy of tumors. PMID- 12905881 TI - [The mouse model and human disease (sequel)]. PMID- 12905880 TI - [Tuberculosis in patients with chronic renal failure undergoing hemodialysis]. AB - OBJECTIVE: To improve the level of early diagnosis and outcome of tuberculosis (TB) in patients with chronic renal failure on regular hemodialysis (HD). METHODS: Thirteen cases among 306 patients undergoing HD from January 1995 to May 2000 were identified, and the clinical presentations, diagnostic procedures and management were analyzed. RESULTS: The incidence of TB in HD patients was 4.2%. The extrapulmonary TB is common, accounting for 76.9%. The main site of TB was pleural in 4 (30.8%) patients. Most of patients were sensitive to anti-TB agents with the total effective rate of 84.6%. CONCLUSIONS: The TB should be diagnosed carefully and treated promptly if suspected in patients with CRF during the period of HD. PMID- 12905882 TI - [Purification of recombinant hEGF expressed in yeast Pichia pastoris and the study on its characters]. AB - OBJECTIVE: To obtain recombinant human epidermal growth factor(hEGF) that can be used in animal experiments and clinical trial. METHOD: Chemically synthesized hEGF gene was expressed in Yeast Pichia pastoris and the secretory hEGF was purified by Phenysepharose 6 Fast Flow(high sub), Q-sepharose High Performance, and Superdex 30 chromatography, and its characters were studied by respective methods. RESULTS: The purified hEGF doesn't contain pyrogen, endotoxin, or yeast chromosome DNA and the purity reached 98%. The recombinant human EGF has correct molecular weight, pI, N-terminal amino acids sequences, peptide map, ultraviolet spectrum and well-biological activity. CONCLUSION: The purified hEGF is in accord with the requirements for animal experiments and clinical trial which provides the basis of preparing EGF agents for clinical test. PMID- 12905883 TI - [Hunting for novel protein factors in G-protein pathway with yeast two-hybrid system]. AB - OBJECTIVE: To explore the protein factors that could interact with G beta subunit within the G protein signal transducing pathway. METHODS: The highly sensitive protein-protein interaction system--Yeast Two-hybrid System was applied to screen the human cDNA library with constructed "Bite plasmid" containing G beta subunit gene fragment. And then the false positive test was adapted. RESULTS: Three positive gene fragments were obtained. One codes for "Actin bundling protein". The other two are new ones and their Gene Bank accession numbers are AF288405 and AF288406 respectively. CONCLUSIONS: It is the first time to find that among human brain tissue, G beta subunit becomes a structural or functional unit interacts with actin bundling protein and the other two unknown protein factors to activate the following pathway. This result may be important to understand the relationship between the actin cytoskeleton and G proteins. PMID- 12905884 TI - [Roles of G-protein beta and gamma subunits in the interaction of G beta gamma with adenylyl cyclases II]. AB - OBJECTIVE: To explore the individual roles of G protein beta and gamma subunit in the interactions with effectors. METHODS: We investigated the interactions of G beta 1 and G beta 1 gamma 2 with adenylyl cyclase II(AC II) using the yeast two hybrid and three-hybrid systems. RESULTS: When assayed for the abilities to activate the reporter gene, the interactions among AD-beta 1, gamma 2 and BD-AC IIQ in the three-hybrid system were more potent than the interactions between AD beta 1 and BD-AC II Q in the two-hybrid system. The expressions of BD-AC IIQ and AD-beta 1 in transformants coexpressed AD-beta 1 and BD-AC IIQ, and transformants coexpressed AD-beta 1, gamma 2 and BD-AC IIQ were respectively detected. The comparisons between the reporter activity and the expression levels of BD-AC IIQ and AD-beta 1 in the yeast cells show there was no correlations, i.e. The difference in the reporter activity was not a reflection of differential expression level of the hybrid proteins. CONCLUSIONS: All these results suggest that G protein beta 1 subunit is sufficient to maintain the basic interaction between G beta 1 gamma 2 and AC IIQ, and gamma 2 subunit plays an important role in the high affinity interaction of G beta 1 gamma 2 with AC IIQ. PMID- 12905885 TI - [Stimulatory effect of chemokine-like factor 1 (CKLF1) on the growth of bone marrow cells]. AB - OBJECTIVE: To investigate the activity of CKLF1 on the proliferation and differentiation of bone marrow cells. METHODS: Human low density bone marrow cells and mouse bone marrow cells were plated in 96-well microplate and supernatants from transfected COS-7 cell culture were added. The cell proliferation was assayed by MTT method after 5 days incubation. The enhancing effect of CKLF1 on the colony formation of human hematopoietic progenitor cells was identified in semi-solid culture. RESULTS: CKLF1 has obvious enhancing effect on both human and mouse bone marrow cells, it can stimulate the colony formation of human hematopoietic stem cells and has synergistic action with GM-CSF. CONCLUSION: CKLF1 can promote the proliferation and differentiation of bone marrow cells. PMID- 12905886 TI - [Expression in E. coli and chemotactic activity analyzing of recombinant CC chemokine RANTES]. AB - OBJECTIVE: To further understand the structure and function of RANTES and provide basis for research for application. METHODS: PCR product corresponding to region encoding mature RANTES protein was cloned into E. coli expression vector; purified recombinant protein was obtained by heparin affinity chromatography and its chemotactic activity was determined by Boyden chamber. RESULTS: The recombinant RANTES was expressed in E. coli with high efficiency. Purified protein showed chemotactic activity to peripheral blood lymphocytes, U937 cells, and CCR4 stable-transfected HEK293 cells. CONCLUSION: The purified recombinant protein exerted chemotactic activity on PBL and cultured cell lines, and we have established a experimental system for further study of structure and function of RANTES. PMID- 12905888 TI - [Differentiation of natural killer cells into two functional subsets: NKh1 and NKh2]. AB - OBJECTIVE: To verify the presence of functional subsets of natural killer cells based on the cytokine production. METHODS: NK cells were purified and cultured in complete RPMI1640 medium in the presence of either IFN gamma + anti-IL 4(classical Th1 polarization) or IL-4 + anti-IFN gamma (classical Th2 polarization) for three days, and then were collected and detected for type I/type II cytokines by RT-PCR method. RESULTS: NK cells were purified from 15 healthy donors, over 70% purity of NK cells were determined by flow cytometry. NK cells in peripheral blood expressed high level of type I cytokines, mainly IFN gamma, but low level of type II cytokines such as IL-10 and IL-13, IL-4 was not produced by NK cells. Cells cultured in IFN gamma + anti-IL-4 condition exhibited significantly increased level of IFN gamma, unchanged IL-2, and decreased type II cytokines. Cells grew in IL-4 + anti-IFN gamma condition exhibited increased IL 10 and IL-13, and decreased IFN gamma expressions. CONCLUSIONS: Based on the cytokine production, NK cells may be divided into two functional subsets in the same manner as that of T lymphocytes(e.g. Th1/Th2): NKh1 and NKh2. The biological characterization and phenotypic marker are under investigate. PMID- 12905887 TI - [Expression and purification of recombinant human interleukin-11 in Pichia pastoris]. AB - OBJECTIVE: To express recombinant human interleukin-11 (rhIL-11) in methylotropic yeast Pichia pastoris. METHODS: By designing and synthesizing an artificial gene for IL-11, the expression vector pPICZ alpha-A-IL-11 was constructed and introduced into Pichia pastoris by linearized electroporation. The rhIL-11 protein was identified by ELISA and SDS-PAGE analysis. The bioactivity was analyzed by B9-11 cell line. A combination of liquid chromatography was developed to purify the rhIL-11 from ferment supernatant. RESULTS: The nucleotide sequence analysis indicated that the sequence of cloned artificial IL-11 gene accorded with that of designed; the secreted yield of rhIL-11 by yeast Pichia pastoris KM71-2424 in flask reached 60 mg/L. The biological activity of IL-11 in yeast supernatant and E. coli standard determined by B9-11 was 5.5 x 10(7) U/mg and 2.2 x 10(7) U/mg respectively. The rhIL-11 was purified to electrophoretic purity by a combination of liquid chromatography. CONCLUSION: The human IL-11 artificial gene was obtained and successfully expressed in the Pichia pastoris(KM71-2424). The biological activity of IL-11 in yeast supernatant was significantly higher than that of E. coli standard. The rhIL-11 was purified to electrophoretic purity. PMID- 12905889 TI - [Toxicity study of epidermal growth factor eye medicine liquid]. AB - OBJECTIVE: To apply for a new medicine certificate, the experiments of acute, long toxicity and side effects of EGF eye medicine liquid were investigated in animals. METHODS: All of examinations were done according to the instructions of "Requirements of the Toxicity Study of New Medicine" RESULTS: No toxicity or side effect of EGF eye medicine liquid given by i.v., i.p., s.c. or eye-drop were been found. CONCLUSION: For the results of whole experiments are in complete accord with "the requirements of the toxicity study of new medicine", EGF eye medicine liquid will be safety in clinical application. PMID- 12905890 TI - [Expression of neurotrophin-4(NT-4) gene in baculovirus expression vector system]. AB - OBJECTIVE: We selected baculovirus expression vector system to express human NT-4 with biological activity. METHODS: The hNT-4 mature peptide-coding sequence is amplified by polymerase chain reaction (PCR), ligated to baculovirus expression vector PacGP67B, and expressed in the insert sf9 cell line. RESULTS: The protein molecular weight of the expressed hNT-4 was about 15,000 by SDS-PAGE, and the hNT 4 antibody could react with this protein in the infected supernatant and total cell by western-blot. The activity of hNT-4 determined by PC12 cell line was definite. CONCLUSIONS: The results may aid for studying the effect of the hNT-4 on basic medical research and clinical application. PMID- 12905892 TI - [The changes of cell structure in the courses of Armillaria mellea penetrating the nutritional stems of Gastrodia elata]. AB - OBJECTIVE: To study the cell structure changes of Gastrodia elata after Armillaria mellea infection as well as the nutritional resource of this important medicinal plant. METHOD: Observation of the serial sections on the nutritional stems of G. elata, and measure new corms of G. elata when cut off the old corms connection with rhizomorph of A. mellea. RESULT: After G. elata nutritional stems infected by A. mellea, the rhizomorph of A. mellea separate into several hyphal layers and penetrate hyphal stream which infect into the cells of cortex layer in the direction of outside and infect directly into layer cells in the direction of inside, these hypha can be used the nutrition of G. elata the new corms will be stop growth if cut off the rhizomorph of A. mellea connected with G. elata. CONCLUSION: After the rhizomorph of A. mellea infected G. elata nutritional stems, hyphal coils and the hyphal stream will breakthrough passage cells as well as large cells surround the whole nutritional stems, so the passage cells is the key nutritional resource in the whole growth period of G. elata. PMID- 12905891 TI - [Eukaryotic expression and in vitro tumor cell apoptosis-induced effect of human TRAIL]. AB - OBJECTIVE: To explore the tumor cell apoptosis induced by the effect in vitro of eukaryotic expression of full-length TRAIL cDNA and its extracellular region. METHODS: The eukaryotic expression vectors for both forms of the cDNA acquired from the fetal heart cDNA library were constructed. After gene transfecting, the stable expression cell lines were obtained by G418 screening. RESULTS: The supernatants from tansfectants could induce apoptosis of different tumor cell lines in vitro, and an enhanced effect was observed by adding TFAR19 (TF-1 cell apoptosis-related protein 19), a novel apoptosis gene product discovered in our laboratory. CONCLUSION: Eukaryotic expression products of TRAIL can induce apoptosis of the tumor cells, and TFAR19 could enhance the effect on apoptosis of tumor cells. PMID- 12905893 TI - [Gene cloning and expression of CTLA-4 in E. coli]. AB - OBJECTIVE: To express hsCTLA-4 in E. coli. METHODS: The hsCTLA-4 gene was obtained by PCR amplification from pE plasmid which contains CTLA-4 gene and was inserted into the expression vector pGEX-2T. The recombination strain was induced by IPTG with different concentrations and time. RESULTS: The sequence of PCR amplified DNA fragments was identical with the reported CTLA-4 gene. SDS-PAGE and Western blot showed that 0.10 mmol/L IPTG can induce higher production of the fusion protein with molecular weight 40,000 after addition of IPTG 4 hours and GST-CTLA4 had immunological activity. CONCLUSIONS: hsCTLA-4 can be expressed in soluble form high efficiency. PMID- 12905894 TI - [Differential expression analysis and cloning of murine thymic aged-related genes]. AB - OBJECTIVE: Differential expression analysis and cloning of murine thymic aged related genes. METHODS: Different expressions of thymic mRNAs from 1- and 10 month old mice were analyzed via DDRT-PCR and different expression sequence tags (ESTs) were obtained, following by identification with Northern Blotting, DNA sequencing, as well as screening of cDNA library. RESULTS: It was found that there would be a significant difference of gene expression in murine thymuses. Gene expression of some genes were exclusive in thymus from 1 or 10-month old mice, while some expressed different with age. 108 differential display cDNA fragments were recovered, among which 31 were positive for hybridization by Northern blot. After sequenced, 14 ESTs were found to share high homology to known genes, whereas remaining 17 were novel. A murine thymic cDNA library was screened by using one cDNA fragment that expressed with higher level in total RNA of 1-month old murine thymic tissues than 10-month old. Finally, one 1,470 bp fragment was cloned and showed a 99% of homology to murine transketolase. CONCLUSION: Expression of murine thymic genes has displayed a marked difference with age. These genes might be participated in thymic atrophy. PMID- 12905895 TI - [The role of neuroimmunoregulation mobilized by enhancing immunologic function via hou hai acupoint antigen injection]. AB - OBJECTIVE: To study the mechanism of neuroimmunoregulation which is triggered by enhancing immunologic function via Hou Hai acupoint antigen injection. METHODS: Immunohistochemical method, immunofluorometric method and RT-PCR were used to examine the different distribution of cytokine immunopositive cells in the brain and expression of cytokines in the spleen of the human IgG sensitized rats received acupoint, subcutaneous and normal rats antigen injections. RESULTS: In the areas of lateral hypothalamic nucleus (LH) and amygdaloid nuclear complex (AA), the distribution of cytokines immunopositive cells with acupoint injection group was significantly increased more than that in the subcutaneous injection group. But the expression of cytokines immunopositive cells both by the acupoint injection and the subcutaneous injection groups reached their peak value in similar time. Double-labelling results showed that the cytokine immunopositive cells were neurons. In the spleens, the expressions of cytokines, IL-2 and IFN, were significantly increased by acupoint injection more than subcutaneous injection. CONCLUSIONS: The time course of neuroimmunoregulation is similar in the immunized rats via both acupoint injection and subcutaneous injection of antigens. But the activity of neuroimmunoregulation is not the same in acupoint and subcutaneous injection groups. Neurons of the LH and AA are the main source of the neuroimmunomodulators. The effect of enhancing immunologic function via Hou Hai acupoint injection is may be more efficient to mobilize the activity of neuroimmunoregulation of immune-associated brain region than modulation of the immune system. PMID- 12905896 TI - [Killing effect of human pulmonary adenocarcinoma cells with TK + CD/5-Fc + GCV coexpression suicide gene systems]. AB - OBJECTIVE: To investigate the different killing effect to human pulmonary adenocarcinoma cell line cells GLC-82 with coexpressed double suicide genes compared with single gene. METHODS: Recombinant expression vectors containing CD (cytosine deaminase) and/or TK (thymidine kinase) gene under CMV promoter were constructed successfully. The vectors were transfected to GLC-82 tumor cell lines by use of lipofectamine. The clones were picked out after G418 selection. Extraneous gene integration and expression were confirmed by PCR and semi quantitative RT-PCR. The cytotoxicity to these transgenic cells under treatment with 5-Fc and GCV were measured by MTT assays. RESULTS: Double and single suicide gene transfer were both stably expressed in GLC-82 cells. The cytotoxic effects of co-expressed TK-CD genes were superior than that of the single gene. CONCLUSION: The CD + TK/5-Fc + GCV co-expression system is more effective for killing effect of tumor cells than CD/5-Fc or TK/GCV system alone. PMID- 12905897 TI - [Weekly cyclophosphamide pulse therapy combined with corticosteroids in the treatment of pemphigus]. AB - OBJECTIVE: To investigate the better regimen of combined cyclophosphamide pulse therapy with corticosteroids in the treatment of Pemphigus. METHODS: Intravenous cyclophosphamide was given weekly in a dosage of 600 mg to those Pemphigus patients whose conditions couldn't be improved by corticosteroids (oral prednisone 1.2-1.5 mg/kg/d) alone. RESULTS: Ten patients with Pemphigus received weekly cyclophosphamide therapy in addition to corticosteroid. Patients conditions improved quickly, without side effects. CONCLUSIONS: Cyclophosphamide weekly pulse therapy combined with corticosteroids is a good regimen in the treatment of Pemphigus. PMID- 12905898 TI - [The basic and applied study on the epidermal growth factor]. AB - This article reviews the results of the basic research about epidermal growth factor and its receptor, and the development of the novel drug, EGF eyedrop, that containing chemically synthesized EGF gene, the construction of EGF expression vector, the transformation of the host cells, the purification of the recombinant protein EGF, the preparation of three batches of the EGF product and identification, the preclinical and clinical trials. Relevant studies show that recombinant EGF consisting of 51 amino acids can be secreted into the medium under the control of the alpha factor leading sequence in the yeast cells. The EGF can accelerate the growth of corneal-limbal epithelial cells and the healing of an alkali burned corneal. The EGF can be used in curing oral cavity ulcer and skin burned wound. And it has the preventive effects on experimental duodenal ulcer of rat. The antiserum was made for test of the concentration of blood EGF and urine EGF by RIA. Data from studies demonstrate the inhibition effect of EGF on the growth of tumor cells, such as A431 and BT325 cells in the presence of high EGF concentration (> 10 ng/ml). The expression of EGFR and DNA ploidy in renal carcinoma has clinical significance. Crystallization and preliminary x-ray diffraction studies of the EGF has been made. The MW of the EGF product is 6000, and the pI is about 4.6 and it has correct N-terminal amino acids sequences, immunogenicity and biological activity. There is no vestige of the DNA of the yeast cells. Animal experiments reveal that there is no cumulation of the EGF in the body, and EGF can promote corneal epithelial healing. There is no toxicological effect during cornea wound healing of rabbit. A randomized, double blind, placebo-controlled, multi-center clinical trial was conducted in four hospitals to assess safety, ocular tolerance and efficacy of an ophthalmic solution of EGF for 200 cases of cornea transplantation and 247 cases of nebulae. Unequivocal results were obtained as the eyedrop really accelerate the wounded cornea healing. So, the EGF eyedrop as a novel drug of class I is approved by the National Drug Administration, and this is the first gene engineering drug that come from yeast expression system in China. PMID- 12905899 TI - [Preliminary investigation on the pathogeny, diagnosis and treatment of chondromalacia patella]. AB - This paper presents the preliminary investigation on chondromalacia patella at our department in recent years. A random cluster sampling survey covering 2743 normal persons was carried out. The prevalence rate is 36.2%. It was found that, applying transmission electron microscope and immunohistochemical methods on to cartilage tissues of the abnormal region, articular cartilage necrosis was in direct proportion with the abnormal pressure, while the restoration capability of local chondrocytes was in inverse proportion with pathological changes and the pressure. The chondromalacia patella was produced by repeated abnormal stress acting on the cartilage. The stress derived from the uncongruency and the decreasing in the contact area of patellofemoral joint when the subluxation or tilt of patellae was caused by the abnormal anatomical and biomechanical relationship. The initial lesion was at the matrix of cartilage, the collagen network was disrupted, then proteoglycan was lost. The microenvironment of chondrocytes was changed with degradation of matrix. So the chondrocytes became degenerative and necrosis from superficial to deep layer, then feed back the matrix again. Finally, the total cartilage layer might disappear, and the bone under cartilage might proliferate. At late stage, the cartilage was completely destroyed and had no self-restorative ability. Therefore, early diagnosis and treatment are necessary. It is highly suggested axis radiograph of the knee with the tibiae tuberositas localization are helpful to early diagnosis. Furthermore, JKY-Muscle Rehabilitation Instrument is invented for non-operative therapy. It enhances muscle power by selective training of the vastus medialis muscle using electrical stimulator to relieve pain and correct subluxation of patella with 90% efficiency (63% of excellent-effective rate). In late stage, patellofemoral replacement is recommended. The excellent-effective rate is 86.3%. PMID- 12905900 TI - [Effects of SDF-1 alpha and its receptor on regulation of hematopoiesis]. AB - Hematopoietic stem/progenitor cell transplantation is one of main choices for treating patients with malignant tumor or heredity diseases in recently year. The homing of hematopoietic stem cells(HSC) is a committed step to ensure the bone marrow hematopoiesis reconstitution. Recent studies have showed that stromal cell derived factor-1 (SDF-1) and its receptor has important effects on proliferation, differentiation, homing of HSC to bone marrow and hematopoiesis reconstitution. The purpose of this review is to give a recent update informations on mechanism of hematopoiesis regulation mediated by SDF-1 and its receptor. PMID- 12905901 TI - [Application of tissue engineering in medicine]. AB - Tissue engineering is a new field, which is based on engineering and life biology. It studies the structure of creatures and shows a wide prospect of treating the functional obstacle of tissues' and organs' wane. Lots of success has been achieved during the passed years and most of them were used on clinical. According to the origin of germinal layer during embryo development, the applying progress of tissue engineering in medicine is discussed. PMID- 12905902 TI - [A study on cellular matrix via cell extraction from bovine pericardium]. AB - OBJECTIVE: To obtain an acellular matrix from bovine pericardial tissue, as a scaffold for tissue engineering of heart valves or cardiovascular patching applications. METHODS: A four step detergent and enzyme link extraction procedure was practiced in this study. The mechanic properties of the acellular matrix were evaluated, and the components were analyzed biochemically. RESULTS: HE staining confirmed the removal of cells and Von Gieson staining showed the integrality of collagen and elastin. Biochemical analysis demonstrated the retention of collagen and some glycosaminoglycans while the percentage of the soluble proteins reduced slightly. The tissue fracture strength and shrinkage temperature made no significant difference after extraction. CONCLUSIONS: The extraction process is effective to remove cells from bovine pericardial tissue while maintains its mechanical strength. This approach may eventually lead to a scaffold for heart valves and cardiovascular patching applications in tissue engineering. PMID- 12905903 TI - [The factors that influence the consistency of international normalized ratio of prothrombin time]. AB - OBJECTIVE: To investigate the factors that influence the consistency of international normalized ratio (INR) of prothrombin time. METHODS: Using the same automated blood coagulation analyzer and INR calibration plasma with two different types of prothrombin time (PT) reagent to assay the INR values of 19 controls (group 1), 12 patients who took long-term anticoagulant orally (group 2), 12 patients who took it at the first time (group 3) and 8 patients who took long-term anticoagulant together with cephalosporin N (group 4). RESULTS: The two kinds of reagents gave quite different INR values in all four groups (P < 0.01). The INR difference in group 3 (0.23 +/- 0.07, n = 12) was larger than that in group 2 (0.51 +/- 0.20, n = 12) (P < 0.01). The difference in group 4 (0.61 +/- 0.21, n = 8) was larger than that in group 2 (P < 0.01). CONCLUSIONS: The factors that may influence INR consistency are the source and sensitivity of the reagents, as well as the period of the anti coagulant been taken and the other drugs taken at the same time. PMID- 12905904 TI - [Effect of recombinant epidermal growth factor on corneal epithelial cells after excision of pterygium]. AB - OBJECTIVE: To investigate the effects of recombinant epidermal growth factor on the wounded corneal healing after excision of pterygium. METHODS: Simple and unrecurrent pterygium was selected, excised and sewed up conjunctival flap under local anaesthesia and microscope. After operation two drops of EGF eyedrop were used, and five minutes later 0.3% Tarivid Eye Oint was used again, and then the operated eye was enswathed. Everyday the wound cornea was observed and the EGF eyedrop was used. RESULTS: The wound cornea healing time of control group was seven days and that of the EGF group was five days and gave a remarkable difference (P < 0.05). The same results were obtained after comparing the effective group and ineffective group, P < 0.05. CONCLUSIONS: EGF eyedrop can accelerate proliferation and recovery of wound corneal epithelial cells. In the clinical trial, every body felt well and no side-effect was observed. PMID- 12905905 TI - [Diagnosis and treatment of one hundred and eighty-nine patients with chronic pancreatitis]. AB - OBJECTIVE: To summarize the experience of the diagnosis and therapy of chronic pancreatitis (CP). METHODS: 189 patients with chronic pancreatitis confirmed surgically from 1983 to Aug 1999 admitted in our hospital were retrospectively studied. RESULTS: Of all 189 patients, the average age was forty-eight years old. The ratio of male to female is 2.1:1. The proportion of confirmed diagnosis to CP is 51.3% within one year and 80.4% within the first five years. The positive rate for diagnosis was 71.9% by CT and 76.9% by ERCP respectively. The positive rate of PABA was 69.7%. While 136(72%) patients received operations, the rate of complication was low. One patient died of infection after pancreatic fistula at the seventh day after surgery, and 97.8% of patients suffered less pain. CONCLUSIONS: The early diagnosis of CP is difficult. As a delicate operation program set at appropriate time is of vital importance for modifying the course of the disease and improving patients' life quality. PMID- 12905906 TI - Correlation between bacterial counts in saliva and subgingival plaque. AB - The aim of this study was to examine the correlation between the number and type of bacteria from periodontal pockets more than 4 mm deep and saliva in 26 patients. Periodontal pocket samples were taken with paper points and transferred to 0.1 ml of enriched thioglicollate broth. Saliva samples were collected simultaneously in aseptic flasks. Both samples were processed within the first hour. They were inoculated in Schaedler agar plus 5 micrograms/ml vitamin K and 5% blood, TSBV agar and MGB agar to perform colony counts and identification. Spirochete counts per microscopic field were obtained by direct light microscopy of Gram-stained preparations. The results show a fair to good correlation between both samples for anaerobic, pigmented gram-negative rods, anaerobic non-pigmented gram-negative rods, spirochetes, facultative gram-negative rods other than Actinobacillus actinomycetemcomitans, anaerobic, gram-positive cocci and anaerobic gram-positive rods (Spearman correlation coefficient 0.51 to 0.96). The correlation coefficient values for A.a., facultative gram-positive rods, facultative gram-positive cocci and facultative gram-negative cocci were lower than 0.21. There were no significant differences between the counts in both samples for all the bacterial groups (Student's t test, p > 0.1). We may conclude that, under the experimental conditions of the present study, saliva samples and periodontal pocket samples are equally useful to detect subgingival organisms associated with periodontal disease in the oral cavity. Saliva samples were useful to evaluate risk and periodontal therapy in individual patients or groups. PMID- 12905907 TI - Iron overloading inhibits dentine mineralization. AB - The present study reveals the inhibitory effect of iron intoxication on the process of dentine mineralization. Wistar rats were injected intraperitoneally with iron dextran at 0.88 g/kg body weight per day for 10 days during the period of odontogenesis. An age-matched group was injected intraperitoneally with bisodium etidronate (EHBP) at 20 mg/kg body weight per day for 10 days. Another age-matched group was treated with similar amounts of saline intraperitoneally and considered as control. At the end of the experimental period the animals intoxicated with iron exhibited non-mineralized dentine and mineralized bone. The animals treated with EHBP showed non-mineralized dentine and bone. These findings would suggest the existence of different mineralization mechanisms for bone and dentine. PMID- 12905908 TI - Quality control of decontaminating agents. AB - The present study evaluates the efficiency of the following decontaminating agents for the multiresistant, locally circulating bacterium Pseudomonas aeruginosa: glutaraldehyde 2%--makes A and B-, glutaraldehyde-formaldehyde; povidone-iodine-makes A, B and C-; sodium hypochloride; chloroxylenol--makes A and B-; and lapire chloride. The 9027 ATCC strain was used as a standard. A modification of the method of Kelsey and Sykes (1) was used to evaluate decontaminating efficiency. Highly satisfactory results were obtained with glutaraldehide 2% A and B, glutaraldehyde-formaldehyde and sodium hypochlorite. The results for povidone-iodine A, B and C were satisfactory but were unsatisfactory for chloroxylenol and lapirium chloride. PMID- 12905909 TI - A case of prepubertal periodontitis and prevalence of gingivitis in a population attending a university clinic in Rosario, Argentina. AB - The present study was based on the finding of a case of prepubertal periodontitis, a low prevalence pathology. One hundred and fifty-two infant patients attending the Department of Odontopediatrics of the Faculty of Dentistry of the National University of Rosario over the period April 1995-September 1996 were included in the present study. The reason for consultation was dental caries. However, the high general prevalence of gingivitis (50%) was remarkable. This may be due to deficient oral health care awareness at home and at school. The prevalence of prepubertal periodontitis (PPP) in the population under study (one case of localized PPP) was 0.66%. These data are in agreement with international reports that account for a prevalence value of less than 1%. A relevant difference with other reported cases could be that this patient complained of upper respiratory tract disorders and, occasionally, ear infections. These symptoms appear in the generalized but not the localized forms of the disease. PMID- 12905910 TI - Histological study of parotid, submaxillary and von Ebner salivary glands in chronic alcoholics. AB - Samples from parotid, submaxillary, and von Ebner salivary glands of six chronic alcoholic individuals who had died of alcoholic hepatic cirrhosis were analyzed by topographic and histochemical routine stains and marked for cytokeratins; two normal adult individuals were used as control. Modifications in the acinar cells were found, but the most evident changes were observed in the ductal system: enlargement of major ducts, heterogeneous expression of cytokeratins and athrophy in epithelial cells, desquamated cells and stasis of content, and ductal hyperplasia in von Ebner glands. The lymphoplasmocytic infiltration does not represent the typical lymphocytic focus on Sjogren's syndrome or other connective tissue pathologies. Our findings indicate that functional and structural variations are produced both in serous acini and ducts parotid, submaxilar and von Ebner glands affected by alcoholic sialosis. PMID- 12905912 TI - Oral mucosa lesions in Mazahua Indian adolescents. AB - The epidemiologic data on oral lesions in native Indians remain unknown in many countries around the world. This paper reports the prevalence and distribution of oral congenital anomalies and pathologic lesions found in a survey of 107 schoolchildren (ages 12 to 17), from two isolated communities in the ethnographic Mazahua area in the State of Mexico. The main entities identified were: pigmented lesions (47.6%), lingual anomalies (17.4%) and developmental tooth alterations (6.9%). The remaining 24.4% of the lesions were gingival inflammatory hyperplasia, partial ankilosis of the tongue, lichen planus, focal epithelial hyperplasia and the double lip. The most frequent localization was lips and tongue. These findings suggest the high prevalence of oral anomalies in this Indian population. Therefore, we suggest that health programs should emphasize the prevention, diagnosis and treatment of these pathologies in Indians groups. PMID- 12905911 TI - An experimental model to study implant corrosion. AB - Corrosion phenomena would appear to play a decisive role in metallic implant long term behaviour. This study presents a method to correlate results obtained in "in vivo" and "in vitro" studies on materials used for metallic implants. Samples of titanium and copper immersed in saline solution (pH 7.4 and 5.2) were used for the "in vitro" study. Implants of these same materials were placed in rat tibiae following the method previously described by Cabrini et al. The animals were killed 14 days post-implantation, the tibiae were resected, radiographed and embedded in acrylic resin. Polarization curves revealed high corrosion in copper implants and low corrosion in titanium implants. It is important to point out that the titanium samples suffered slightly higher corrosion when immersed in a lower pH medium (pH 5.2), as in chronic inflammatory processes. A passive film was found on titanium samples as opposed to the strong corrosion observed in the copper implants. Microscopy revealed osseointegration around titanium implants and a severe inflammatory reaction with abscesses surrounding the copper implants. The method presented herein would allow to correlate "in vivo" and "in vitro" studies on corrosion in different implant materials and establish their relation with cell response. PMID- 12905913 TI - Efficacy of two techniques for endodontic instrumentation. AB - The aim of the present study was to compare the quality of preparation in curved and straight root canals employing 2 techniques, i.e. the step-back technique and the technique of Goerig, Michelich and Schultz. One hundred and thirty single rooted human teeth were employed throughout. The walls of the root canals of these teeth were painted. Following instrumentation the roots were sectioned longitudinally. The sections were observed under a stereoscopic microscope SZ40 with a grid fitted in the eyepiece. The efficacy of instrumentation was evaluated using area of paint removed as the end-point. Statistical significance of the data was tested with the chi square test employing the Epi Info 5 software. There was no statistically significant difference between techniques for straight canals. The technique of Goerig, Michelich and Schultz was more effective than the step-back technique in the apical third of curved canals. PMID- 12905914 TI - Oral health of children from rural excluded villages (Trancas and Calchaqui valleys. Tucuman, Argentina). AB - The general hypothesis of the study was to evaluate strategically important rural areas to confirm their characteristics of "excluded population" and identify the priorities to allocate human and financial resources of the international co operation project. The aim of this study was to establish the degree of development of villages in the Trancas and Calchaqui valleys, and the oral health of children living in areas involved in an integral study on excluded villages in the northwest of Argentina. These villages are considered a priority in the UNIR Project; they are thus strategically important as a link with countries of the MERCOSUR. Data on living conditions were obtained through direct inquiries (Project UNIR, W. K. Kellogg Foundation/University of Tucuman) and from data supplied by the Ministry of Economy, by the Ministry of Culture and Education and by the SIEMPRO project of the Secretariat of Social Development of the Ministry of Health. The Human Development Index (HDI) was calculated as World Bank criteria (1995), The studies on dental conditions were carried out in all the children between 12 and 14 years old, that is a total of 58 in the Trancas Valley and 212 in the Calchaqui Valley. RESULTS: (1) The HDI was 0.731 for the province, 0.56 for the Trancas Valley and 0.51 for Calchaqui valley. (2) The results are above the values for the country in the first case, and below the average country values for the valleys. (3) The percentage of BNN families reached 92%. The results for oral health show: (4) the DMFT were 6.34 +/- 0.07 in the Trancas Valley and 7.47 +/- 0.276 in the Calchaqui valley. (5) Carious teeth (CT) were the most important component of this index. (6) The absence of a specific and preventive assistance. CONCLUSIONS: (a) Children between 12 and 14 living in the Valleys of Trancas and Calchaqui (Tucuman, Argentina) are affected by severe levels of caries, with values above the national and province means. (b) The human development index is below that of the corresponding national and province levels. (c) Inhabitants of these valleys can be identified as socially and biologically highly vulnerable. (d) The indicators of cariogenic risk reveal the importance of introducing oral preventive care components to the health plan for the area focused on the excluded population. PMID- 12905915 TI - Preventive action of short-term and long-term chlorhexidine rinses. AB - The purpose of this study was to evaluate the effects of 0.12% chlorhexidine mouthrinses on the presence of plaque and gingivitis, either used as the only oral hygiene practice during a short period of time (21 days) or associated with routine daily brushing for 180 days. The experimental subjects were 40 dental students of an average age of 20.9 years. Following a thorough examination and mechanical prophylaxis. The subjects were divided into two treatment groups. Twenty subjects stopped all oral hygiene procedures for 21 days except for twice daily rinses with 0.12% chlorhexidine or a placebo. Plaque, gingivitis scores and oral soft tissue health were recorded on days 0, 7, 14 and 21 for the short term group and on days 0, 90 and 180 of experimentation for the long term group. After 21 days of rinsing, plaque accumulation, gingivitis and bleeding on probing were significantly lower in the chlorhexidine group than in the placebo group. After three and six months of mouthrinses, the chlorhexidine group had less plaque and significantly less gingivitis than the placebo group. No significant differences in adverse oral soft tissue effects were observed between the different groups. It was concluded that a 0.12% chlorhexidine digluconate mouthrinse can provide an important adjunct to the prevention and control of gingivitis when used as the only oral hygiene practice or with regular personal hygiene procedures. The results obtained in this clinical study, using stringent parameters of evaluation with statistic analysis, corroborated the results obtained by other studies. PMID- 12905916 TI - Beyond "bad news": the diagnosis, prognosis and classification of lymphomas and lymphoma patients in the age of biomedicine (1945-1995). PMID- 12905917 TI - To kill or not to kill: the eradication of contagious bovine pleuro-pneumonia in western Europe. PMID- 12905918 TI - Part of a scientific master plan? Paul Ehrlich and the origins of his receptor concept. PMID- 12905919 TI - More than just a propagandist for tea: religious argument and advice on a healthy life in the work of the Dutch physician Cornelis Bontekoe (1647-1685). PMID- 12905920 TI - Illustrations from the Wellcome collections. Medicine man: the forgotten museum of Henry Wellcome. PMID- 12905921 TI - Is the NCS diet obsolete? PMID- 12905922 TI - Quality reports. CMS and JCAHO strive to help consumers make better healthcare decisions. PMID- 12905923 TI - A venue for theological/ethical issues. PMID- 12905924 TI - Telling the values story at Catholic Health East. PMID- 12905925 TI - Service can be the best marketing tool. PMID- 12905926 TI - Can medical schools be Catholic? For two reasons, Catholic universities should continue sponsoring such schools. PMID- 12905927 TI - Creating a medical futility policy. Physicians at a Honolulu hospital describe the experience. PMID- 12905928 TI - Are futility policies the answer? Caregivers must improve communication with patients and their families. PMID- 12905930 TI - Baton Rouge's "virtual clinic". Physicians and dentists have pooled their skills to provide care for the area's working poor. PMID- 12905929 TI - Transforming nursing. A Wisconsin hospital and its nurses have discovered the power of partnership. PMID- 12905931 TI - Leveraging the power of the proxy. Catholic health care systems are using stock holdings to push for corporate accountability. PMID- 12905932 TI - Lessons of mission discernment. The process renders decision making more participatory and transparent and produces better decisions, too. PMID- 12905934 TI - Portraits in collaboration. A bishop takes the lead in Wichita. PMID- 12905933 TI - Cases in genetics, Part 4. Lack of consent is an ethical issue. PMID- 12905935 TI - [Progress of hematology analyzers in Japan]. PMID- 12905936 TI - [Advanced technologies of new automated hematology analyzers]. PMID- 12905937 TI - [Clinical usefulness of automated hematology analyzer system for WHO classification of hematological neoplasia]. PMID- 12905938 TI - [Automation and systematization in hemostatic assays: now and future]. PMID- 12905939 TI - [Circulating hematological malignant cells in peripheral blood, not detected by automated hematology analyzer]. PMID- 12905940 TI - [Standardization of clinical flow cytometric analysis]. PMID- 12905942 TI - [Automation for blood smear preparation]. PMID- 12905941 TI - [Tips on monoclonal antibody reagents for flow cytometry (appendix: CD chart 2003)]. PMID- 12905943 TI - [Automation for blood transfusion examination]. PMID- 12905944 TI - [In vitro microbiology tests and crisis management in hospital]. PMID- 12905945 TI - [Chemical system Synchron LX20 pro]. PMID- 12905947 TI - [Capillary zone electrophoresis and its clinical application]. PMID- 12905946 TI - [Access immunoassay system--troponin I and myocardial damage]. PMID- 12905948 TI - [Total laboratory automation system]. PMID- 12905949 TI - [Total laboratory system: Quality control]. PMID- 12905950 TI - [International standardization: collaboration with NCCLS]. PMID- 12905951 TI - [Automated systems for nucleic acid tests by polymerase chain reaction]. PMID- 12905952 TI - [Chromosomal analysis, fluorescence in situ hybridization method, and spectral karyotyping in hematologic malignancies]. PMID- 12905953 TI - [Gene testing and clinical laboratory]. PMID- 12905954 TI - [A prospect of genetic tests]. PMID- 12905955 TI - [Globalization of clinical laboratory-present status and future--ISO/TC212 and JCTLM]. PMID- 12905956 TI - [POCT--practice of cooperation in medical care]. PMID- 12905957 TI - [Proper recognition of the laboratory testing is keenly needed to keep the meaning for existence of both hospital in-house laboratories and commercial referral laboratories]. PMID- 12905958 TI - [The role and the future of medical technologists]. PMID- 12905959 TI - [Analysis of animal material]. PMID- 12905960 TI - [Papers dedicated to the 90th Birthday of Tat'iana Antonovna Detlaf]. PMID- 12905961 TI - [Research on chemical constituents of Lonicera: a review and prospects]. PMID- 12905962 TI - Intermittent infusions of zoledronic acid are as effective as daily bisphosphonates in increasing bone mineral density in port-menopausal women. PMID- 12905963 TI - Long-term treatment with diacerein has a structure/disease modifying effect on hip osteoarthritis. PMID- 12905964 TI - PASSCLAIM - Report of First Plenary Meeting including a set of interim criteria to scientifically substantiate claims on foods. PMID- 12905966 TI - [Introduction to the main topic]. PMID- 12905965 TI - [Great attention should be paid to preclinical and clinical research of venous diseases in China]. PMID- 12905967 TI - [Considerations from a medical view]. PMID- 12905968 TI - [The DRG-System and its consequences especially on geriatrics]. PMID- 12905969 TI - [The necessity of quality development in the care of dementia patients and their relatives]. PMID- 12905970 TI - [The hot point in management of cerebrovascular disease]. PMID- 12905971 TI - [New conception of antihypertension treatment: endothelium-nitric oxide-oxidation stress chain]. PMID- 12905972 TI - 2 diseases + 1 shot = 10 years of protection. PMID- 12905973 TI - HPNA educational resources for nursing assistants. PMID- 12905974 TI - [Prosthetic surgery of the knee--a success story to be continued]. PMID- 12905975 TI - [Towards research into hepatitis B virus infection]. PMID- 12905976 TI - [Some problems in HLA typing in bone marrow register/cord blood bank]. PMID- 12905977 TI - Plantar wart treatment with combination imiquimod and salicylic acid pads. AB - Treatment of plantar warts is often difficult and may be painful, often employing destructive treatment modalities. We report the successful treatment of a patient with a large plantar wart using Imiquimod 5% cream under occlusion with a 40% salicylic acid pad. This combination treatment modality likely allows successful delivery of Imiquimod through the thick skin on the plantar surface. Once penetrated, an anti-viral state is created by upregulating specific cytokines to eradicate the human papilloma virus (HPV). PMID- 12905978 TI - Helicobacter pylori management: how to improve the therapeutic confusion in practice. AB - There appears to be a disconnect between current guidelines for Helicobacter pylori testing and treatment, and clinical practice, including physician beliefs and actual prescribing patterns. In particular, there are markedly different approaches in primary and secondary care, and country-specific differences in eradication therapy for H pylori infection. Although most physicians do not believe that H pylori causes non-ulcer dyspepsia, the majority appear to prescribe eradication. Less information is available on the management of H pylori infection and gastro-esophageal reflux disease, and more marked differences in attitudes and practice occur in this condition. Even in peptic ulcer disease, where most clinicians both in primary and in secondary care believe H pylori should be eradicated, there is often a breakdown in the translation of this belief into practice. There is also confusion in terms of treatment regimens applied for H pylori eradication. Eradication regimens are less successful in practice than in clinical trials. Furthermore, a sizable proportion of patients with peptic ulcer remain symptomatic despite cure of the ulcer diathesis, which may undermine confidence. Therapeutic confusion about what to prescribe, side effects limiting compliance, bacterial resistance, and socioeconomic factors may all impair therapeutic success with eradication therapy in practice. Unfortunately, it has been well documented that guidelines alone are likely to have little or no impact in practice. Publication in a journal is unlikely to lead to effective implementation in primary care. On the basis of available evidence, clinical behaviour is most likely changed when guidelines are developed by the peer group of clinicians for whom they were intended, are disseminated through a specific educational program, and are implemented by applying, preferably during the consultation, specific reminders. PMID- 12905979 TI - Is it time for quadruple therapy to be first line? AB - The most commonly used regimen for Helicobacter pylori therapy at present is twice-daily proton pump inhibitor (PPI)-based triple therapy. Bismuth-based therapy is the next most common treatment used by gastroenterologists. When a PPI is combined with bismuth-based triple therapy (quadruple therapy), eradication rates are increased as compared with the triple therapy alone. Three separate randomized trials from three continents that compare quadruple therapy and PPI based triple therapy revealed remarkably similar results. Eradication rates with PPI-based triple therapy and quadruple therapy were not significantly different. The eradication rates with quadruple therapy were 3% to 6% higher than PPI triple therapy, indicating that quadruple therapy should be no less effective than PPI triple therapy. Furthermore, these two therapies had similar rates of compliance and adverse events. The major potential benefit of the quadruple therapy relates to antibiotic resistance. In patients with clarithromycin resistance, PPI-based triple therapy, but not quadruple therapy, had a significantly lower eradication rate. However, due to its ability to largely overcome metronidazole resistance, quadruple therapy had little if any decrement in eradication rates compared with PPI triple therapy inpatients with metronidazole-resistant H pylori. Therefore, quadruple therapy can be considered a first line therapy for H pylori. PMID- 12905981 TI - [Policy paper on primary prevention of cardiovascular diseases. Current draft of 3/25/2003 issued by the Executive Committee of the German Society of Cardiology, Heart and Circulation Research. Edited by the Prevention Project Group on behalf of the Executive Committee]. PMID- 12905982 TI - [Stem cell sources of hepatic cells]. PMID- 12905980 TI - [Guidelines for diagnosis and treatment of chronic coronary heart disease. Issued by the executive committee of the German Society of Cardiology--Heart Circulation Research in cooperation with the German Society for Prevention and Rehabilitation of Cardiac Diseases and the German Society for Thoracic and Cardiovascular Surgery]. PMID- 12905983 TI - [Complementary medicine in Exeter: a 10-year jubilee]. PMID- 12905984 TI - [10 years research in complementary medicine. A bridge between past and future. Interview with Prof. Dr. h.c. Alois Stacher, Vienna, by M. Ullmann]. PMID- 12905985 TI - Executive insights. Joseph F. Corfits, Jr., Senior Vice President of Finance and CFO, Central Iowa Health System. PMID- 12905988 TI - Scope-adjusted impact factor. PMID- 12905989 TI - [Report on the 9th Magdeburg Work Conference, 25-26 October 2002. "Palliative oncological surgery and second tumors"]. PMID- 12905990 TI - [Should one screen and treat condyloma lesions in the male partner of a female patient with a CIN? It is not useful to screen and treat]. PMID- 12905992 TI - [Prominent Russian neurosergeon and transfusionist (on the 120th birthday of academician V N Shamov]. PMID- 12905991 TI - [Tendon replacement operations on the foot]. PMID- 12905993 TI - [Veniamin Mikhailovich Ugriumov (on his 90th birthday)]. PMID- 12905995 TI - Current awareness in prenatal diagnosis. PMID- 12905994 TI - The mitochondrial permeability transition pore complex as a pharmacological target. An introduction. PMID- 12905996 TI - [Proceedings of the 3d All-Russian Scientific Conference on "Persistence of microorganisms". Orenburg, September 2000]. PMID- 12905997 TI - Issues in outpatient PPS: keys to successful revenue cycle management. PMID- 12905999 TI - "DNA theft": new crime in the UK. PMID- 12906000 TI - Data trends. Peer comparisons promote improved billing practices. PMID- 12905998 TI - [On the 90th birthday of Tat'iana Antonovna Detlaf]. PMID- 12906001 TI - Fortunes rising! Results of the 2003 HFMA Compensation Survey. PMID- 12906002 TI - American Society of Clinical Oncology-39th Annual Meeting. Taxanes. 31 May-3 June 2003, Chicago, IL, USA. PMID- 12906003 TI - American Society of Clinical Oncology-39th Annual Meeting. Angiogenesis. 31 May-3 June 2003, Chicago, IL, USA. PMID- 12906004 TI - American Society of Clinical Oncology-39th Annual Meting. HRPC and NSCLC. 31 May 3 June 2003, Chicago, IL, USA. PMID- 12906005 TI - American Society of Clinical Oncology-39th Annual Meeting. Investigational Drugs. 31 May-3 June 2003, Chicago, IL, USA. PMID- 12906006 TI - Digestive Disease Week 2003. 17-22 May 2003, Orlando, Fl, USA. PMID- 12906007 TI - Digestive Disease Week 2003. Crohn's disease, constipation and IBS. 17-22 May 2003, Orlando, Fl, USA. PMID- 12906008 TI - Finished Y-chromosome sequence reveals genomic surprises. PMID- 12906009 TI - American Society for Microbiology-103rd General Meeting. 18-22 May 2003, Washington DC, USA. PMID- 12906010 TI - Tracking spongiform encephalopathies in North America. PMID- 12906011 TI - Australian and New Zealand Society of Nuclear Medicine-33rd Annual Scientific Meeting. 10-13 May 2003 Sydney, Australia. PMID- 12906012 TI - Monkeypox spreads as US public-health system plays catch-up. PMID- 12906013 TI - Immunology-15th European Congress. 8-12 June 2002, Rhodes, Greece. PMID- 12906014 TI - Clinical Trials in Cancer-SMi Conference. 11-12 June 2003, London, UK. PMID- 12906016 TI - Pain therapeutics-SMi Conference. 11-12 June, 2003, London, UK. PMID- 12906017 TI - SARS in hiding: WHO calls for vigilance. PMID- 12906018 TI - Capecitabine: new indication. In breast cancer: inadequately assessed. AB - There is no consensus on treatment of locally advanced or metastatic breast cancer after failure of first-line cytotoxic chemotherapy. Common options are continuous infusion of a taxane (docetaxel or paclitaxel), vinorelbine or fluorouracil. Capecitabine is now licensed for use in breast cancer, both in combination with IV docetaxel after anthracycline failure, and as single-agent therapy after failure of anthracyclines and taxanes. The clinical evaluation dossier on capecitabine fails to answer the most important questions about comparative efficacy and safety. In second-line treatment, after anthracycline failure, the only available comparative trial showed that the capecitabine + docetaxel combination increased median survival time by about three months relative to placebo + docetaxel, but caused more adverse events. There are no trials comparing capecitabine with other options. There is no evidence that capecitabine increases the length or quality of survival, relative to intravenous vinorelbine, in women with breast cancer that is resistant to both anthracyclines and taxanes. The classical adverse effects of capecitabine are also observed in women with breast cancer, namely palmoplantar erythrodysesthesia, diarrhoea, nausea and vomiting, and major hyperbilirubinemia. Capecitabine can be taken by mouth and this may be an advantage. However, current evidence is too limited to justify using capecitabine outside of clinical trials. PMID- 12906019 TI - Rituximab: new indication. In aggressive non Hodgkin lymphoma: benefits must be confirmed. AB - The first-line treatment for diffuse large-B-cell non Hodgkin's lymphoma, a highly malignant lymphoma, is CHOP chemotherapy (cyclophosphamide + doxorubicin + vincristine + prednisone). Rituximab, a monoclonal antibody targeting certain B cells, has received a new indication in the treatment of this type of lymphoma, in combination with the CHOP protocol. In late 2002, the only available evaluation data came from one comparative, unblinded trial in patients over 60 years of age. Addition of rituximab to the CHOP protocol increased both the overall two-year survival rate (70% versus 57%), and the two-year event-free survival rate. Other trials are underway. In this trial, 9% of patients had major systemic reactions during the first rituximab infusion (respiratory disturbances, chills, fever and hypotension). These reactions did not occur during subsequent infusions. About 6% of patients had serious cardiac arrhythmias. In practice, the CHOP protocol remains the standard treatment for aggressive non Hodgkin's lymphoma. Pending further information, addition of rituximab to the CHOP protocol may be justified for patients who meet the inclusion criteria used in the only available clinical trial. PMID- 12906020 TI - Etanercept: new preparation. Useful after methotrexate failure in inflammatory rheumatism. AB - There is no reference second-line treatment for patients with rheumatoid arthritis, juvenile chronic arthritis, psoriatic arthropathy or ankylosing spondylitis after failure or intolerance of a slow-acting antirheumatic drug such as methotrexate. Etanercept, a immunosuppressant targeting TNF-alpha (like infliximab), is now approved in France for use in these situations, with the exception of spondylitis. In the second-line treatment of adults with rheumatoid arthritis, the clinical evaluation dossier on etanercept contains data from dose finding studies and two placebo-controlled trials involving patients in whom several single-agent treatments had failed. At a dose of 25 mg subcutaneously twice a week, etanercept worked partially in about half the patients. Without direct comparisons, the place of etanercept relative to other slow-acting antirheumatic drugs is difficult to establish. From indirect comparisons, etanercept seems a slightly better treatment option than infliximab. In the first line treatment of rheumatoid arthritis, one trial showed that etanercept worked faster than methotrexate, but there was no significant difference between the two treatments after two years. Little is known about the efficacy of etanercept in patients with juvenile chronic arthritis who do not respond adequately to methotrexate. There are no comparative trials. One double-blind placebo controlled trial showed that etanercept, when it worked, remained active for at least 7 months. In one trial, etanercept was more effective than placebo in patients with psoriatic arthropathy and ankylosing spondylitis who continued to receive their usual treatment, which included a slow-acting antirheumatic drug in about 50% of cases. More than 50% of patients treated with etanercept have a cutaneous reaction to the injection. These reactions are usually mild or moderate. Active pharmacovigilance is needed, given its mechanism of action, and previous notifications of a wide variety of adverse effects (even though it is sometimes difficult to establish a foolproof link between etanercept and the adverse effect). Long-term studies of large numbers of patients are needed to determine the precise risk of side effects including haematological, infectious, neurological, oncological and immunological effects. In practice, methotrexate remains the first-line treatment for inflammatory arthritis. Etanercept can be a useful second-line treatment, especially in juvenile chronic arthritis. PMID- 12906021 TI - Valganciclovir: new preparation. CMV retinitis: a simpler, oral treatment. AB - For AIDS patients with cytomegalovirus (CMV) retinitis, the standard initial treatment has so far been ganciclovir or foscarnet, both of which are infused intravenously. The choice between these two antiviral agents is based on their differing side effects. Maintenance treatment is based on ganciclovir or foscarnet, but both these drugs must be given as daily intravenous infusions. Oral ganciclovir is less effective. Intraocular maintenance treatment in appropriate only for patients with localised infections and a low risk of relapse. Valganciclovir can be taken orally for both initial and maintenance treatment of CMV retinitis. Valganciclovir has good bioavailability and is rapidly metabolised into ganciclovir. A comparative randomised trial in 160 patients showed that oral valganciclovir is as effective as intravenous ganciclovir when used as an initial treatment. There are no comparative trials of oral valganciclovir as a maintenance treatment, but non comparative data and pharmacokinetic studies suggest that oral valganciclovir is about as effective as intravenous ganciclovir. The adverse effects of oral valganciclovir are similar to those of intravenous ganciclovir, except that the oral route avoids the risk of local complications at the infusion site. Valganciclovir causes more frequent diarrhoea and oral candidosis than intravenous ganciclovir. In practice, for patients with CMV retinitis who require systemic treatment, oral valganciclovir is easier to use and, in our opinion, should now replace IV ganciclovir for both initial and maintenance treatment. The convenience of oral valganciclovir should also limit the use of purely intraocular treatment of CMV retinitis. PMID- 12906022 TI - Hepatitis B immunoglobulin: new preparation. Prolongs survival after liver transplantation. AB - The main complication of liver transplantation for hepatitis B is viral re infection of the graft. It occurs in 50-100% of patients in the absence of prevention, and reduces survival. The clinical evaluation dossier on hepatitis B immunoglobulin contains data from three retrospective studies. Hepatitis B immunoglobulin reduced the rate of graft infections and increased the survival time (measured between 2 and 5 years after grafting) by 15-30% in absolute terms. Treatment should probably be maintained indefinitely with an intravenous injection of 10,000 IU every two to three months, in order to keep anti-HBs antibody titres at adequate levels. In patients with signs of active hepatitis B virus replication, hepatitis B immunoglobulin alone cannot prevent graft infection. Such patients should receive also lamivudine. Hypersensitivity reactions during infusion are the main adverse effects. Hepatitis B immunoglobulin should now be standard treatment for preventing relapse after liver transplantation for hepatitis B. PMID- 12906023 TI - Paroxetine: new indication. In social phobia: minimal assessment. AB - Social phobia is generally defined as an intense and persistent fear of one or several social situations, with important repercussions for occupational activity or social life. Cognitive psychotherapy and antidepressants have partial efficacy. There is no reference drug therapy. In France, paroxetine is the first drug to be granted a licence for patients with social phobia. Clinical evaluation consists of data from four placebo-controlled trials lasting only 12 to 24 weeks. Treatment with paroxetine was associated with a significant improvement in standard social-phobia scores, although most patients remained symptomatic. Paroxetine is the best assessed selective serotonin reuptake inhibitor in this setting. However, long-term data are lacking, and the disorder is chronic. Paroxetine has not been compared with cognitive therapy. About one-third of patients in clinical trials stopped taking paroxetine, mainly because of adverse events. Gastrointestinal upset, sleep disturbance and ejaculatory problems are frequent. Paroxetine also has the potential to interact with other drugs. In practice, paroxetine may help some patients, provided they are aware of its limitations. The long-term effects of paroxetine in this setting remain unknown. PMID- 12906025 TI - Bibliography. Current world literature. Retina and vitreous disorders. PMID- 12906024 TI - Conversations with Sally Ride. Interview by Linda Voss. PMID- 12906027 TI - Plasma levels of IL-8 predict early complications in patients with coronary heart disease after percutaneous coronary intervention. AB - The aim of the present study was to investigate the prognostic value of plasma interleukin-8 (IL-8) for early complications after percutaneous coronary intervention (PCI). The pre- and postprocedural plasma levels of IL-8 and serum C reactive protein (CRP) were examined by immunoassay, and the expression of CD11b/CD18 on neutrophils was assessed by flow cytometry. Early complications (abrupt occlusion, threatened abrupt occlusion, early recurrence of ischemia, myocardial infarction, cardiac sudden death, and target vessel revascularization) occurred intra-procedure and 30 days after PCI and were observed in 121 consecutive patients with coronary heart disease. Sixteen patients with early complications had high preprocedural levels and high postprocedural differentials of IL-8, CRP, and CD11b/CD18 compared to those without complications (all P < 0.05). The occurrence of complications showed a significant increase in the patients according to the tertiles of IL-8, CRP, and CD11b/CD18. Preprocedural levels of IL-8 (RR = 5.864, CI = 1.658-20.734, P = 0.006) and diabetes (RR = 1.587, CI = 1.246-2.132, P = 0.038) were independent predictors of early complications. There were significant correlations in the postprocedural differential between IL-8 and CD11b/CD18 (r = 0.776, P = 0.002) in patients with complications. The results reveal that the early complications after PCI contribute to preprocedural inflammatory responses. Normal levels of IL-8 may be powerful negative predictors of early complications in patients with CHD following PCI. PMID- 12906026 TI - DNA double helix hits 50. PMID- 12906028 TI - Effect of trimetazidine on exercise performance in patients with coronary artery ectasia. AB - Coronary artery ectasia (CAE) is a rare form of coronary artery disease. It has previously been shown that nitrate derivatives induce exertional angina in patients with CAE. Furthermore, there is limited data about the effects of other anti-ischemic agents in CAE. The aim of this study was to investigate the effect of trimetazidine on exercise performance in patients with CAE. The study population consisted of 56 patients with isolated CAE (32 males, mean age: 58 +/- 9 years). The presence of myocardial ischemia was evaluated by treadmill exercise test. The exercise test was positive in 49 patients at baseline and in 27 patients during trimetazidine therapy (P < 0.01). The exercise test induced angina in all of the patients who have had a positive test result. Significant ST depressions were observed in 42 and 23 patients before and after the treatment, respectively (P < 0.01). The extent of ST depression at peak exercise was significantly lower on trimetazidine (0.9 +/- 0.5 vs 1.5 +/- 0.6 mm, P < 0.01). With trimetazidine, the exercise duration increased from 7.8 +/- 2.1 to 8.7 +/- 2.4 min (P = 0.04) and cardiac work-load also increased from 8.9 +/- 2.3 to 10.4 +/- 2.1 mets (P < 0.01). The results suggest that trimetazidine can relieve exercise-induced angina and improve exercise performance in patients with CAE. PMID- 12906029 TI - Different long-term course between chest pain and exercise-induced ST depression in syndrome X. AB - The aim of the present study was to assess the long-term clinical course of patients with syndrome X, focusing on different courses between exercise-induced ST depression and chest pain. Forty-three patients with syndrome X were followed up for 6.4 +/- 3.8 years. They were divided into the 3 groups according to chest pain: disappeared (n = 24), improved (n = 14), or unchanged (n = 5). No patients had cardiac events and all had a favorable long-term prognosis. In patients showing disappearance of chest pain, exercise-induced ST depression and rate pressure product (RPP) at peak exercise did not change during follow-up. However, ST depression and RPP decreased significantly in those with improved chest pain. These observations suggest that abnormal pain perception plays an important role in the development of chest pain. PMID- 12906030 TI - Factor V Leiden mutation and its relation to left atrial thrombus in chronic nonrheumatic atrial fibrillation. AB - The genetic defect of coagulation factor V, known as factor V Leiden, produces a resistance to degradation by activated protein C and increased venous thrombosis. However, the role of factor V Leiden in the formation of left atrial thrombus with nonrheumatic atrial fibrillation has not been studied. We investigated whether factor V Leiden is a risk factor for left atrial thrombus in patients with nonrheumatic atrial fibrillation. We analyzed clinical, echocardiographic, and biochemical data in 105 consecutive patients with nonrheumatic atrial fibrillation. These patients were divided into two groups: group A (n = 37) with left atrial thrombus and group B (n = 68) without left atrial thrombus. The study also included 42 control subjects. Left atrial thrombus was investigated by using both transthoracic echocardiography and transesophageal echocardiography. Blood samples from the patients and controls were analyzed for the factor V Leiden mutation by DNA analysis, using the polymerase chain reaction. There was no significant difference in the prevalence of factor V Leiden between the patients and control subjects. The prevalence of factor V Leiden mutation was 8.1% (3/37) in patients with left atrial thrombus, and 8.8% (6/68) in patients without left atrial thrombus. The prevalence of factor V Leiden was 7.1% (3/42) in control subjects. The prevalance of factor V Leiden was 10% (2/20) in patients with spontaneous echo contrast and 8% (7/85) in patients without spontaneous echo contrast. Multivariate analyses showed that left ventricular ejection fraction was an independent predictor of left atrial thrombus. Factor V Leiden mutation is not a risk factor for left atrial thrombus formation and spontaneous echo contrast in patients with nonrheumatic atrial fibrillation. PMID- 12906031 TI - Endogenous opioids and epinephrine in nitroglycerin provocation tilt test in patients with neurally mediated syncope. AB - Endogenous opioids and catecholamines are involved in autonomic activity. Nitroglycerin provocation tilt is a useful modality for evaluating neurally mediated syncope. Endogenous opioids and epinephrine might play an important role in nitroglycerin provocation tilt. To investigate whether or not opioids and catecholamines are involved in the pathogenesis of nitroglycerin provocation tilt, we measured the temporal changes of the plasma levels of beta endorphin, norepinephrine, and epinephrine in 64 patients with syncope of unknown etiology, and compared the findings with those of 16 patients who underwent isoproterenol provocation tilt (1-3 microg/min) test with a positive response. We performed a 20 minute control tilt (80 degrees) followed by a nitroglycerin provocation tilt of 20 minutes with the intravenous infusion of nitroglycerin. Nitroglycerin infusion was started at 250 microg/h, and was increased by 250 microg/h every 3 minutes up to 1500 microg/h during the tilt test. Beta-endorphin, norepinephrine, and epinephrine were measured in peripheral venous blood in the supine position 2, 10, and 20 minutes after the start of the tilt test, and also at the onset of syncope. Twenty-six patients had a positive response to the control tilt (group 1), and 22 patients had a positive response to nitroglycerin provocation tilt (group 2). The remaining 16 patients had a negative response to both control tilt and nitroglycerin provocation tilt (group 3), compared with isoproterenol provocation tilt patients (group 4). Beta-endorphin and epinephrine only significantly increased in groups 1 and 2 (beta-endorphin; from 7.3 +/- 3.3 pg/mL to 19.9 +/- 17.7 pg/mL, in group 1, P < 0.05; from 7.3 +/- 2.9 to 16.5 +/- 10.7 pg/mL, in group 2, P < 0.05; epinephrine; from 42 +/- 58 pg/mL to 157 +/- 161 pg/mL, in group 1, P < 0.05: from 33 +/- 25 to 202 +/- 252 pg/mL, in group 2, P < 0.05), but not in groups 3 and 4. Beta-endorphin and epinephrine might participate in the pathophysiology in conventional tilt-induced as well as nitroglycerin provocation tilt-induced syncope in patients with neurally mediated syncope. PMID- 12906032 TI - Cardiac muscle cell disorganization in apical hypertrophic cardiomyopathy: a cardiac biopsy study. AB - Apical hypertrophic cardiomyopathy has been divided into two entities: apical asymmetric septal hypertrophy (apical ASH) and apical symmetric hypertrophy (AH). The latter differs clinically from hypertrophic cardiomyopathy (HCM) with ASH, and it is unclear whether AH represents a distinct subtype of HCM. In the present study, the presence or absence and the extent of cardiac muscle cell disorganization, a histologic characteristic of HCM, were compared in patients with AH (n = 10) and ASH (n = 29) in whom cardiac biopsy specimens were obtained from the left ventricular apex and interventricular septum. Disorganization was graded as (1+) in only 1 patient in the AH group and (-) in the remaining 9. In contrast, in the ASH group disorganization was graded as (1+) in 15 patients, (2+) in 7, (3+) in 3, and (-) in only 4 (P < 0.0001). Thus, it was observed that in AH disorganization is virtually absent or at most limited to a very narrow area. It is concluded from a histological stand point as well that the type of apical hypertrophic cardiomyopathy showing apical symmetric hypertrophy differs from usual HCM. PMID- 12906033 TI - Relationship between impaired chronotropic response, cardiac output during exercise, and exercise tolerance in patients with chronic heart failure. AB - The present study was undertaken to investigate the relationship between the extent of impaired chronotropic response and cardiac output during exercise, and exercise tolerance in patients with chronic heart failure. The subjects consisted of 24 patients (mean 60.1 +/- 14.0 years) who had mild chronotropic incompetence. Cardiopulmonary exercise testing was performed in all patients, and heart rate (HR), anaerobic threshold (AT), maximum oxygen uptake (peak VO2), slope of the regression line relating the ventilatory equivalent to carbon dioxide output (VE/VCO2 slope), and exercise time were measured. Cardiac output (CO) was measured by a thoracic bioimpedance method and cardiac index (CI) was calculated. Plasma norepinephrine (NE) was measured at rest and immediately after the exercise test. The changes in HR, NE, and CI from the resting state to immediately after exercise were calculated as deltaHR, deltaNE, and deltaCI, respectively. The deltaNE was converted to a logarithmic scale and deltaHR/log deltaNE was used as a parameter of HR response to sympathetic nerve stimulation. The results were as follows: HR and NE in the resting state had no correlation with AT and with peak VO2. DeltaHR/log deltaNE correlated positively with both AT and peak VO2, and negatively with the VE/CO2 slope. DeltaHR/log deltaNE correlated positively with peak CI, %deltaCI, and deltaCI/exercise time. The data suggest that one of the mechanisms of low exercise tolerance in chronic heart failure patients was due to an inadequate increase in CO response against exercise caused by an impaired HR response to increased NE. PMID- 12906034 TI - Leptin might be a regulator of serum uric acid concentrations in humans. AB - Increased serum urate concentration is a frequent finding in patients with hypertension. Since hyperuricemia is associated with obesity, renal disease, hyperlipidemia, and atherosclerosis, whether or not serum urate is a cardiovascular risk factor per se has remained elusive. The subjects were 210 Turkish male and 210 female adults over 20 years of age. None had diabetes mellitus, endocrine diseases, or renal or hepatic disease, and those receiving antihypertensive drugs, systemic corticosteroids, or lipid-lowering drugs were excluded. Height, weight, blood pressure, serum glucose, lipid profiles, serum insulin, DHEA-SO4, and leptin were measured in the morning after an overnight fast. Women had significantly higher mean leptin (20.3 +/- 0.88 ng/mL vs 5.78 +/- 0.39 ng/mL, P < 0.001) and lower mean uric acid (248.03 +/- 4.76 micromol/L vs 311.6 +/- 5.35 micromol/L, P < 0.001), triglyceride (1.42 +/- 0.06 mmol/L vs 1.61 +/- 0.06 mmol/L, P < 0.001), and DHEA-SO4 (3.02 +/- 0.17 micromol/L vs 4.43 +/- 0.19 micromol/L, P < 0.001) concentrations than men, even when adjusted for BMI. On univariate correlation analysis, leptin showed the strongest association with BMI in both sexes and also correlated significantly with BMI, insulin, uric acid, glucose, total cholesterol, and triglycerides in males and BMI, insulin, uric acid, total cholesterol, apo B, and creatinine in females after adjustment for age and BMI. A statistical model containing creatinine, leptin, insulin, and triglycerides accounted for 34% of the variance in serum uric acid levels in men, whereas another consisting of creatinine, triglycerides, leptin, SBP, and insulin explained 42% of the variance in serum uric acid in women. The present study suggests that leptin could be one of the possible candidates for the missing link between obesity and hyperuricemia. Our study may also suggest that hyperuricemia is not only a metabolic end product but also a marker of a major pressor or pathogenic mechanism underlying the hypertension in obesity. PMID- 12906035 TI - Effects of angiotensin-converting enzyme inhibition on changes in left ventricular myocardial creatine kinase system after myocardial infarction: their relation to ventricular remodeling and function. AB - We assessed the effects of angiotensin-converting enzyme (ACE) inhibition on changes in the myocardial intracellular creatine kinase (CK) system in relation to left ventricular (LV) remodeling and function in heart failure after myocardial infarction (MI) in rats. We compared the findings at 4 weeks after MI to those at 12 weeks after MI. LV weight and chamber size were significantly increased and percent fractional shortening (%FS) was decreased in untreated MI rats compared with normal control animals both at 4 and 12 weeks after MI. Animals with MI and treated with the ACE inhibitor temocapril showed significantly reduced LV weight and chamber size and increased %FS compared with untreated MI rats at 12 weeks after MI, but not at 4 weeks after MI. At 4 weeks after MI, no significant changes were found in the total creatine and relative distribution of each CK isoenzyme in either the temocapril-treated or untreated animals with MI compared with the normal controls. In contrast, at 12 weeks after MI, untreated MI rats showed significant reductions in the total creatine and mitochondrial and MM-CK fractions and increases in the MB- and BB-CK fractions compared with the controls. The alterations in the mitochondrial and MB-CK fractions were significantly attenuated after 12 weeks of ACE inhibition. Thus, LV myocardial energy metabolism is progressively impaired and its alteration is not related to the magnitude of geometric changes and LV dysfunction after MI. Most of the beneficial effects of ACE inhibition were observed at 12 weeks after MI. Our results may provide an insight into the therapeutic strategy of ACE inhibition in chronic heart failure after MI. PMID- 12906036 TI - Norepinephrine-induced inositol 1,4,5-trisphosphate formation in atrial myocytes is regulated by extracellular calcium, protein kinase C, and calmodulin. AB - We investigated whether alteration of extracellular and intracellular Ca2+ concentrations, protein kinase C, and calmodulin modulate norepinephrine (NE) induced inositol 1,4,5-trisphosphate (IP3) formation in neonatal rat atrial myocytes. NE-induced IP3 production in atrial myocytes was stimulated by elevation of extracellular Ca2+ in a dose-dependent manner. However, TMB-8 (an intracellular calcium antagonist) and A23187 (an intracellular calcium agonist) did not significantly affect NE-induced IP3 production. PMA (a protein kinase C agonist) significantly decreased and staurosporine (a protein kinase C antagonist) significantly stimulated NE-induced IP3 production. W7 (a calmodulin antagonist) significantly increased the NE-induced IP3. In conclusion, elevation of extracellular Ca2+ concentrations affects NE-induced IP3 formation in atrial myocytes. Protein kinase C and calmodulin may control the IP3 response to NE by a negative feedback mechanism. PMID- 12906037 TI - Spontaneous intimal dissection in a patient with post-infarct angina: identification with intravascular ultrasound and treatment with coronary stenting. AB - A 45-year-old Turkish male patient was admitted to our hospital for an acute myocardial infarction. He had suffered two previous acute myocardial infarctions 5 and 6 years earlier. Coronary angiography performed after the previous two acute myocardial infarctions had shown normal coronary arteries and coronary vasospasm had been suspected. The patient was treated with thrombolytic therapy (rt-PA) during the last coronary event. Five days after the current admission, the patient had postinfarct angina and underwent coronary angiography, showing only a nonobstructive lesion with irregular ulcerated edges in the left anterior descending artery after the first diagonal branch. Subsequent intravascular ultrasound confirmed the presence of an atherosclerotic lesion with plaque dissection. Stenting of this lesion was performed with successful relief of on going chest pain. Intravascular ultrasound can provide important diagnostic information in patients presenting with acute coronary syndromes in the absence of severe angiographic stenosis. Identification of plaque dissection at mildly stenotic lesions provides pathophysiologic insights that may have therapeutic implications. However, the optimal treatment for such lesions is not known. PMID- 12906038 TI - Side branch protection with hydrophilic polymer coated guide wire during cutting balloon angioplasty of a bifurcated lesion. AB - Cutting balloon angioplasty (CBA) was performed in a patient with in-stent restenosis (ISR) which had an important side branch. We used a hydrophilic polymer-coated guide wire for side branch protection during CBA. After CBA was successfully performed, the cutting balloon and guide wire were microscopically examined and proven to have suffered minor damage which, in itself, did not disturb the procedure. Hydrophilic polymer-coated wire might be an effective and safe choice for ISR which needs to be treated by CBA while protecting an important side branch. PMID- 12906039 TI - Reversible ischemic neurological deficit (RIND) due to exercise testing for the diagnosis of angina pectoris. AB - A 57 year old Japanese male with chest oppression due to exercise is presented. This symptom was likely due to effort angina pectoris. Master's double two-step test revealed ischemic ST segment depression on the electrocardiogram, thus, a Tl201 myocardial stress imaging test using a bicycle ergometer was undertaken. Immediately following the exercise test, the patient experienced dizziness and palsy in his left upper and lower limbs. Cerebral angiography demonstrated 70% stenosis at the right internal carotid artery, but no abnormal findings were demonstrated on a cranial x-ray CT scan and magnetic resonance imaging. His palsy in the left limbs completely recovered within 3 weeks. Thus this event was diagnosed as a reversible ischemic neurological deficit (RIND). Major but non cardiogenic complications during exercise testing are very rare, and RIND has not been reported thus far to the best of our knowledge. PMID- 12906040 TI - Acute aortic dissection developing acute myocardial infarction diagnosed by accidentary inserting the catheters into the false lumen during catheterization: a pitfall and rare image. AB - A case of acute aortic dissection complicating acute inferior myocardial infarction diagnosed by accidentally inserting the catheters into the false lumen during the emergency cardiac catheterization is reported. Although the incidence of acute aortic dissection developing acute myocardial infarction is not very rare, an aortogram and coronary angiogram via the false lumen has never been seen before. This case also illustrates how myocardial infarction can mask aortic dissection. Cardiologists should pay more attention to this severe complication of acute aortic dissection and perform transthoracic echocardiography prior to catheterization or fibrinolysis. PMID- 12906041 TI - Evidence for anti-ischemic effect of dual-chamber pacing in patients with the obstructive form of hypertrophic cardiomyopathy. AB - Dual-chamber pacing reportedly improves the quality of life by reducing the frequency of anginal episodes in selected patients with the obstructive form of hypertrophic cardiomyopathy (HCM), although the underlying mechanism or coronary effect is poorly understood. We report 3 patients with obstructive HCM, in whom the effects of atrial vs. dual-chamber tachypacing on systemic hemodynamics and myocardial lactate metabolism were studied. In all patients myocardial lactate production, objective evidence of myocardial ischemia, was demonstrated during atrial pacing, whereas no patient developed myocardial ischemia during dual chamber pacing. By contrast, the responses of pressure gradient to pacing varied among the patients. These observations demonstrate for the first time that dual chamber pacing exerted an anti-ischemic effect in obstructive HCM, which may contribute, at least partly, to the beneficial effects of chronic AV pacing on angina status and/or LV function. PMID- 12906042 TI - Kappa-casein interactions in the suspension of the two major calcium-sensitive human beta-caseins. AB - The possible effects of both the beta-casein (beta-CN) phosphorylation level and the kappa-CN glycosylation level on micelle formation were studied using the doubly-phosphorylated form (beta-CN-2P) and the quadruply-phosphorylated form (beta-CN-4P) of human beta-CN, along with bovine kappa-CN to compare with previous studies using the more highly glycosylated human kappa-CN. Addition of bovine kappa-CN to human beta-CN-2P, beta-CN-4P, or a 1/1 (wt/wt) mixture of the two was at kappa/beta molar ratios from 0.0 to approximately 0.6 and micelles were reconstituted by addition of Ca+2 either directly at 37 degrees C for determination of the fraction suspended or at an initial temperature of 4 degrees that was gradually increased to 37 degrees C with the change in particle size monitored by turbidity measurements. Analysis of the data indicates that the 4P form requires more kappa-CN for stabilization than the 2P form but that the mixture of the two is more like the 4P form in that lateral kappa-kappa interactions may enhance beta-kappa interactions and micelle formation. Above a kappa/beta molar ratio of about 0.2, the caseins were fully suspended into reconstituted micelles. However, micelle size decreased at a higher ratio, indicating that the kappa-CN probably occupies a surface position and may regulate micelle size by its relative abundance. A comparison with published results suggests that the higher glycosylation level of human kappa-CN may protect a larger surface area and result in smaller micelles. Changes in reconstituted micelle size with pH indicate that positively charged groups in the kappa-CN may interact with the negatively charged phosphate esters in the beta-CN moieties in addition to kappa-beta hydrophobic interactions. PMID- 12906044 TI - Lysinoalanine content of formulas for enteral nutrition. AB - Casein and caseinates are the main ingredients of formulas for enteral nutrition. Their manufacturing procedure and the thermal treatments necessary to assure microbiological stabilization and satisfactory shelf-life of the end-products are particularly favorable for the formation of lysinoalanine (LAL), a cross-linked amino acid that is considered a useful marker of the thermal damage and reduced digestibility of proteins. The lysinoalanine content of 18 different kinds of formulas for enteral nutrition was determined by HPLC after derivatization. The liquid formulas have an average value of 528 microg/g protein LAL, ranging from 160 to 800 microg/g protein (average content of formulas for pediatric use 747 microg/g protein). These values are rather high considering that the average value detected in UHT-treated drinkable milk is 117 microg/g protein. In principle, the preparation of caseinates and the thermal stabilization of the end products are the two steps more favorable for the formation of LAL. The fact that the five samples stabilized by an UHT-treatment have an average value of 512 microg/g protein suggests that the LAL content depends more on the quality of the starting ingredients than on the sterilization process. A better selection of the starting ingredients should improve the quality of formulas for enteral nutrition, which is very desirable when formulating foods for consumers with very high nutritional demands. PMID- 12906043 TI - Pseudomonas aeruginosa lectin PA-IIL as a powerful probe for human and bovine milk analysis. AB - Milk composition exhibits species-specific differences depending on genetic, evolutionary, and environmental factors. In addition, commercial milk preparations are also changed by industrial manipulations, including severe heat processing. Cow milk, used as human food, provides important nutrients but lacks some essential components that are present in raw human milk. The present study, which was aimed at comparing infant breastfeeding to cow-based formula nourishment, shows major differences between the human and the commercial cow milk glycans detectable by the lectins PA-IL (galactose-binding) and PA-IIL (fucose and mannose-binding) isolated from the cells of human pathogen Pseudomonas aeruginosa. More than 40 human milk samples, several cow milks, and bovine milk-based infant formulas, were examined using these two lectins. For purposes of comparison, the plant lectins Concanavalin A (Con A), which binds mannose, and Ulex europaeus 1st lectin (UEA-I), which binds fucose, were also used. The most prominent difference was revealed using PA-IIL, which displayed a unique high sensitivity to the human milk fucosylated compounds. PA-IL and UEA-I also exhibited preferential sensitivity to the human milk but considerably lower than that of PA-IIL. Con A was inhibited by human and the other milk preparations examined to the same extent. These findings indicate the superb applicability of PA-IIL for rapid and reliable comparative investigation of milk glycans from human and cow, indicating which glycans could be added to infant formulas in order to enrich them, as well as for verification and quality control of otherwise improved bovine milk-based infant formulas. PMID- 12906045 TI - Selective enumeration of Lactobacillus delbrueckii ssp. bulgaricus, Streptococcus thermophilus, Lactobacillus acidophilus, bifidobacteria, Lactobacillus casei, Lactobacillus rhamnosus, and propionibacteria. AB - Nineteen bacteriological media were evaluated to assess their suitability to selectively enumerate Lactobacillus delbrueckii ssp. bulgaricus, Streptococcus thermophilus, Lactobacillus casei, Lactobacillus rhamnosus, Lactobacillus acidophilus, bifidobacteria, and propionibacteria. Bacteriological media evaluated included Streptococcus thermophilus agar, pH modified MRS agar, MRS vancomycine agar, MRS-bile agar, MRS-NaCl agar, MRS-lithium chloride agar, MRS NNLP (nalidixic acid, neomycin sulfate, lithium chloride and paramomycine sulfate) agar, reinforced clostridial agar, sugar-based (such as maltose, galactose, sorbitol, manitol, esculin) media, sodium lactate agar, arabinose agar, raffinose agar, xylose agar, and L. casei agar. Incubations were carried out under aerobic and anaerobic conditions at 27, 30, 37, 43, and 45 degrees C for 24, 72 h, and 7 to 9 d. S. thermophilus agar and aerobic incubation at 37 degrees C for 24 h were suitable for S. thermophilus. L. delbrueckii ssp. bulgaricus could be enumerated using MRS agar (pH 4.58 or pH 5.20) and under anaerobic incubation at 45 degrees C for 72 h. MRS-vancomycine agar and anaerobic incubation at 43 degrees C for 72 h were suitable to enumerate L. rhamnosus. MRS vancomycine agar and anaerobic incubation at 37 degrees C for 72 h were selective for L. casei. To estimate the counts of L. casei by subtraction method, counts of L. rhamnosus on MRS-vancomycine agar at 43 degrees C for 72 h under anaerobic incubation could be subtracted from total counts of L. casei and L. rhamnosus enumerated on MRS-vancomycine agar at 37 degrees C for 72 h under anaerobic incubation. L. acidophilus could be enumerated using MRS-agar at 43 degrees C for 72 h or Basal agar-maltose agar at 43 degrees C for 72 h or BA-sorbitol agar at 37 degrees C for 72 h, under anaerobic incubation. Bifidobacteria could be enumerated on MRS-NNLP agar under anaerobic incubation at 37 degrees C for 72 h. Propionibacteria could be enumerated on sodium lactate agar under anaerobic incubation at 30 degrees C for 7 to 9 d. A subtraction method was most suitable for counting propionibacteria in the presence of other lactic acid bacteria from a product. For this method, counts of lactic bacteria at d 3 on sodium lactate agar under anaerobic incubation at 30 degrees C were subtracted from counts at d 7 of lactic bacteria and propionibacteria. PMID- 12906046 TI - Concentration of polar MFGM lipids from buttermilk by microfiltration and supercritical fluid extraction. AB - Buttermilk contains the milk fat globule membrane (MFGM), a material that possesses many complex lipids that function as nutritionally valuable molecules. Milk-derived sphingolipids and phospholipids affect numerous cell functions, including regulating growth and development, molecular transport systems, stress responses, cross membrane trafficking, and absorption processes. We developed a two-step method to produce buttermilk derivative ingredients containing increased concentrations of the polar MFGM lipids by microfiltration and supercritical fluid extraction (SFE). These processes offer environmentally benign alternatives to conventional lipid fractionation methods that rely on toxic solvents. Firstly, using a ceramic tubular membrane with 0.8-micron pore size, we evaluated the cross flow microfiltration system that maximally concentrated the polar MFGM lipids using a 2n factorial design; the experimental factors were buttermilk source (fresh, or reconstituted from powder) and temperature (50 degrees C, and 4 degrees C). Secondly, a SFE process using supercritical carbon dioxide removed exclusively nonpolar lipid material from the microfiltered buttermilk product. Lipid analysis showed that after SFE, the product contained a significantly reduced concentration of nonpolar lipids, and a significantly increased concentration of polar lipids derived from the MFGM. Particle size analysis revealed an impact of SFE on the product structure. The efficiency of the SFE system using the microfiltration-processed powder was compared much more favorably to using buttermilk powder. PMID- 12906048 TI - Effect of feeding prepubertal heifers with a high oil diet on mammary development and milk production. AB - The objective of this study was to evaluate the effect of feeding prepubertal heifers a diet containing a high level of polyunsaturated fatty acids on mammary development and milk production. A total of 116 Holstein heifers were either fed a conventionally formulated concentrate or a high oil (HO) concentrate, using the same formulation but including 20% soybean oil, from birth to 6 mo of age. After 6 mo of age, all heifers were managed identically. Mammary gland development was evaluated on heifers slaughtered at 4 mo (n = 10) and 12 mo (n = 30) of age. Other heifers were bred when they reached 15 mo of age and milk production and feed intake were recorded every day from wk 4 to 18 of lactation. Feeding the high oil concentrate increased the concentration of linoleic acid in blood plasma (176%) and mammary fat pad (78%) at 4 mo of age and mammary fat pad (93%) at 12 mo of age. At 4 mo of age, mammary development was similar in both treatments. At 12 mo of age, total, parenchyma, and stroma weights of the mammary gland were not affected by treatments. However, lipid content was lower and concentration of DNA was higher in the parenchyma of heifers fed the high oil diet. Nevertheless, total parenchymal DNA and dry fat free tissue content did not reach statistical significance despite the fact that they were, respectively, 15 and 21% higher in HO heifers. Milk production and composition was not affected by treatments. In conclusion, feeding prepubertal heifers with a high oil concentrate slightly improved the mammary development but effects were too small to be translated into better lactating performances. PMID- 12906049 TI - Effect of intramammary Escherichia coli endotoxin in early- vs. late-lactating dairy cows. AB - We investigated the differences in the effect of intramammarily infused endotoxin for cows in early (EL) and late (LL) lactation. In this crossover study, nine cows were challenged twice with 100 microg of intramammarily infused Escherichia coli 0111:B4 LPS, each cow serving its own control. Systemic and local signs were recorded throughout the experiment to assess the severity of each cow's response. Daily milk yield and indicators of inflammation in blood and milk were also recorded. The response was significantly more severe in the EL period. Before the challenge, milk yield and serum free fatty acid concentration were higher and serum urea concentration lower in EL compared with LL cows. No significant differences were present in other parameters. After the challenge, milk SCC and leukocyte function, measured as chemiluminescence, increased, more markedly in EL. Blood neutrophil content varied from neutropenia to neutrophilia. Simultaneously with neutropenia, the function of neutrophils increased in LL but decreased in EL. Serum cortisol peaked 4 h after challenge in both groups. In conclusion, endotoxin challenge resulted in more severe response in EL. During that time cows also had impaired neutrophil function postchallenge. PMID- 12906050 TI - L-selectin and beta2-integrin expression on circulating bovine polymorphonuclear leukocytes during endotoxin mastitis. AB - The aim of this in vivo study was to examine the effect of intramammarily administered endotoxin (lipopolysaccharide, LPS) on the expression of L-selectin (CD62L) and the beta2-integrin subunits CD11b and CD18 on circulating bovine PMN. Six early lactating cows were infused with Escherichia coli LPS. The adhesion molecules under study were stained at the cell surface and analyzed flow cytometrically. In addition, some of the clinical parameters associated with adhesion molecule mobilization such as fever, blood cortisol levels, somatic cell count (SCC), and total and differential blood leukocyte count were measured. In analogy with observations during clinical coliform mastitis, a progressive decrease of CD62L expression levels was observed early after LPS infusion, concomitantly with a continuous rise of CD11b and CD18 density. However, no correlation was found between the kinetics of CD11b and CD18 density. The initial changes in adhesion molecule expression paralleled the decrease in blood PMN numbers, together with the increase in rectal temperature, cortisol levels, SCC, and number of circulating immature PMN. In conclusion, intramammarily administered LPS seems to play an important role in modulating adhesion receptor expression on circulating bovine PMN. Interestingly, in contrast to coliform mastitis, the net CD18 variation is not principally influenced by CD11b upregulation during endotoxin administration. The knowledge of adhesion molecule kinetics in relation to the different parameters evaluated in the present study contributes to an improved understanding of the inflammatory reaction. PMID- 12906047 TI - Relationships among milk yield, body condition, cow weight, and reproduction in spring-calved Holstein-Friesians. AB - Relationships among milk production, body condition score (BCS), body weight (BW), and reproduction were studied using logistic regression on data from 6433 spring-calving Holstein-Friesian dairy cows in 74 commercial herds. Multivariate models were adjusted for herd, breeding value for milk yield, proportion of Holstein-Friesian genes, lactation number, calving period, and degree of calving assistance. Significant associations between reproductive measures and components of energy balance were identified. Higher 200-d milk protein content and higher protein-to-fat ratio at start of breeding were associated with increased likelihood of submission for breeding in the first 21 d of the breeding season (SR21). High 100-d cumulative milk yield as a proportion of estimated 305-d milk yield (low persistency) was associated with a lower likelihood of pregnancy to first service (PREG1), whereas cows reaching peak milk yields earlier tended to have higher PREG1. Cows that reached nadir milk protein content relatively late in lactation had lower PREG1. Milk yield at first service and 305-d milk protein content were positively associated with the likelihood of pregnancy after 42 d of breeding (PR42). Higher 305-d milk lactose content was associated with increased PREG1 and PR42. Mean BCS at 60 to 100 d of lactation was positively associated with both SR21 and PR42, whereas nadir BCS was positively associated with PREG1. Cows with precalving BCS > 3.0 that also lost > 0.5 BCS unit by first service had lower PR42. More BW gain for 90 d after start of breeding was associated with higher SR21 and PREG1; more BW gain for 90 d after first service was associated with higher PR42. Milk protein and lactose content, BCS, and BW changes are important tools to identify cows at risk of poor reproduction. PMID- 12906052 TI - Effect of omitting one milking weekly on lactational performances and morphological udder changes in dairy cows. AB - The response of dairy cows to omitting one milking weekly was investigated in two successive experiments conducted with Holstein cows milked twice daily. Experiment 1 compared the lactational performances and udder changes in eight cows (31.2 L/d, 201 d in milk) in the 5 wk before and the 5 wk after introducing the suppression of one milking weekly. Milk yield was recorded daily and milk composition twice weekly. Milk partitioning in the udder (alveolar and cisternal milk) and cisternal size (ultrasonography), 8 h after milking, were also measured at the start and the end of the experiment. Although daily milk yield decreased 32% during the experiment (10 wk), linear regression analysis revealed a loss of milk yield of 1.1 L/d (3.7%) as a consequence of the omission of one milking weekly. Milk composition, lactation persistency, and somatic cell count (SCC) were unaffected by milking omission. Milk partitioning in the udder decreased by 38% in alveolar milk volume and showed a tendency to decrease in cisternal milk volume (15%) and cisternal size (7%), as a result of milking omission and advancing lactation. Loss in total milk yield was negatively related with cisternal milk volume (r = -0.77) and cisternal size (r = -0.70) indicating smaller losses in the udders with large cisterns. In Experiment 2, five cows (21.0 L/d, 227 d in milk) previously adapted to the milking omission schedule were used to study the daily effects of milking omission on milk yield, milk composition and udder health during 10 wk. Milk yield and milk composition were approximately constant but SCC increased with lactation stage. The omission of one milking caused an important decrease in milk yield, fat content and SCC on the omission day and a compensatory increase over the following 2 d, but milk protein and lactose did not vary. All variables reached the average weekly value three days after the milking omission (six milkings). In conclusion, under the conditions used, omitting one milking weekly slightly reduced milk yield and did not affect milk composition when healthy cows were used. Milk losses by milking omission depend on udder cistern characteristics; evaluating cistern size by ultrasonography may be a useful tool for choosing cows that are better adapted to a reduced milking frequency. PMID- 12906051 TI - Manipulation of antioxidant status fails to improve fertility of lactating cows or survival of heat-shocked embryos. AB - Experiments were conducted to test whether enhancement of antioxidant status could improve fertility and milk yield in dairy cows and resistance of cultured embryos to heat shock. Three experiments in three herds were performed to evaluate the effect of multiple intramuscular injections of 500 mg of vitamin E and 50 mg of selenium at 8 to 21 d before expected calving and at 30 and 80 d postpartum on reproduction of lactating Holstein cows. Vitamin E and selenium injections did not improve reproductive function or milk yield in any of the studies. The predicted 305-d milk yield (averages of least-squares means across treatments) were: 9478, 7073, and 10,204 kg projected 305-d milk for experiments 1, 2, and 3, respectively. Percentages of cows pregnant at first service were 30, 16, and 24% in experiments 1, 2, and 3, respectively. Three studies were performed to test whether vitamin E improved development of cultured bovine embryos exposed to heat shock. Heat shock of 41 degrees C at the two-cell stage reduced development to the blastocyst stage, but culture with 100 microM vitamin E did not reduce effects of heat shock on impaired development. For example, 9 h at 41 degrees C reduced blastocyst development from 51.2 +/- 3.3% to 3.4 +/- 3.3% in the absence of vitamin E and from 54.0 +/- 3.3% to 5.2 +/- 3.3% in the presence of vitamin E. Development of morulae to the blastocyst stage was not compromised by culture at 41 degrees C for 9 h. Additionally, there was no overall effect of vitamin E on morula development. In conclusion, multiple injections of vitamin E and selenium at the administered levels did not improve postpartum fertility nor milk yield of lactating Holstein cows in three different herds, and there was no direct thermoprotective effect of vitamin E for cultured, heat-shocked embryos. PMID- 12906054 TI - Short communication: Tissue distribution of leptin and leptin receptor mRNA in the bovine. AB - Detection of leptin and leptin receptor mRNA in various tissues is crucial to an understanding of leptin physiology in dairy cattle. We report here evidence of leptin receptor gene expression in central and peripheral tissues of the bovine by reverse transcription and polymerase chain reaction analysis. Leptin mRNA was detectable in mammary parenchyma and in adipose tissue with similar transcript abundance among the subcutaneous, pericardial, perirenal, and mesenteric adipose depots. The mRNA for the long-form of the leptin receptor, Ob-Rb, was detectable in all four adipose depots, mammary parenchyma, semintendinosus muscle, liver, adrenal cortex, spleen, kidney, testis, mesenteric lymph node, lung, aorta, abomasum, duodenum, jejunum, ileum, hypothalamus, pituitary, brain stem, cerebral cortex, cerebellar cortex, pons, and pineal gland. The mRNA for the short form of the leptin receptor, Ob-Ra, was detectable in the liver, adrenal cortex, spleen, pituitary, and brain stem, but not in the other tissues surveyed. The wide spectrum of tissues expressing the leptin receptor gene reveals that leptin may have multiple physiological functions in the bovine. PMID- 12906053 TI - Effects of the mammary gland on functional capacities of blood mononuclear leukocyte populations from periparturient cows. AB - The composition and functional capacity of peripheral blood mononuclear leukocyte populations from dairy cows are altered substantially during the peripartal period. These changes are associated with a heightened susceptibility of the mammary gland to infection. It has been postulated that the metabolic demands associated with lactogenesis may impact negatively leukocyte function during the periparturient period. In the present study, serum immunoglobulin G1 concentration and functional capacities of peripheral blood mononuclear leukocytes from intact (n = 6) and mastectomized (n = 6) periparturient Jersey cows were evaluated and compared. Cell function assessments included lymphocyte proliferation, immunoglobulin M secretion, and interferon-gamma secretion by unstimulated and pokeweed mitogen stimulated mononuclear leukocytes. Data were summarized as mean responses for 5-d periods beginning 21 d prepartum and concluding at 19 d postpartum. The progressive decrease in serum immunoglobulin G in intact but not mastectomized cows before parturition likely was attributable to the selective uptake of this isotype by the mammary gland. Lymphocyte proliferation and secretion of interferon-gamma and polyclonal IgM by mitogen stimulated leukocytes from intact cows decreased during the 15-d period before calving, reaching a nadir at 0 to 4 d postpartum. From 5 to 19 d postpartum, these functions often were comparable to those observed 2 to 3 wk prepartum. Functions of leukocytes from mastectomized cows did not change during the study period, although they often were of lower magnitude than those of cells from nonlactating cows. These results reconfirm the occurrence of a generalized reduction in blood mononuclear leukocyte function during the periparturient period. They also suggest that the reduction in leukocyte function during the period may be, in part, due to the physiologic demands imposed on the dairy cow by the lactating mammary gland. PMID- 12906055 TI - Differential expression of the lactose transporter gene affects growth of Staphylococcus aureus in milk. AB - In this study, differential display polymerase chain reaction (PCR) was used to search for unique or enhanced expression of genes in prevalent bovine mastitis causing Staphylococcus aureus strains. Comparison of a pair of prevalent and rare strains revealed the differential expression of several genes. The lactose specific permease, enzyme II (EII), was highly expressed in the prevalent strain. This gene was selected for further study due to its potential influence on bacterial growth, because lactose is the primary carbohydrate in milk. Growth analysis illustrated that prevalent strains reach significantly higher growth densities sooner than rare strains. Quantitative competitive reverse transcription PCR (QC RT-PCR) revealed increased EII mRNA expression in prevalent strains as compared to rare strains. Mutation of the EII gene resulted in abrogated growth and decreased EII mRNA expression in media containing lactose. These data suggest that increased EII expression may facilitate the pathogenesis of S. aureus mastitis by enhancing growth. This study is the first to implicate EII as a potential virulence factor in mastitis, and therefore may be useful in the development of novel therapeutic strategies against S. aureus mastitis. PMID- 12906056 TI - Elevated milk soluble CD14 in bovine mammary glands challenged with Escherichia coli lipopolysaccharide. AB - The purpose of this study was to determine whether soluble CD14 (sCD14) in milk was affected by stage of lactation, milk somatic cell count (SCC), presence of bacteria, or lipopolysaccharide (LPS)-induced inflammation. Milk samples from 100 lactating cows (396 functional quarters) were assayed for sCD14 in milk to determine effects of stage of lactation, SCC, and intramammary infection. The concentration of sCD14 was highest in transitional milk (0 to 4 d postpartum) and in milk with high SCC (> 750,000 cells/ml). Most of the infected quarters (> 80%) were infected by coagulase-negative staphylococci and yeast. No difference was found between noninfected and infected quarters. One quarter of six healthy lactating cows was challenged with 100 microg LPS in order to study the kinetics of sCD14 during an LPS-induced inflammation. Milk samples were collected at various intervals until 72 h after injection. Rectal temperature, milk tumor necrosis factor-alpha, and interleukin-8 increased immediately after challenge. The increase in sCD14 paralleled the increase in SCC, peaked at 12 h, and started to decline after 24 h. Serum leakage, as characterized by the level of bovine serum albumin in milk, peaked at 4 h and then gradually decreased. All parameters remained at basal levels in control quarters throughout the study. In vitro experiments indicated that neutrophils released sCD14 in response to LPS in a dose-dependent manner. The results indicate that the concentration of sCD14 was significantly increased in milk after LPS challenge. The increase was not likely due to serum leakage. Instead, infiltrated neutrophils might be the main source of increased sCD14 in milk during inflammation. PMID- 12906057 TI - Effects of long chain fatty acids on lipid and glucose metabolism in monolayer cultures of bovine hepatocytes. AB - The objectives were to determine the long-term (48 h) effects of specific long chain fatty acids on hepatic lipid and glucose metabolism in monolayer cultures of bovine hepatocytes. From 16 to 64 h after plating, hepatocytes from three 7- to 10-d-old calves were exposed to one of the following treatments: 1 mM palmitic acid (1 mM C16:0), 2 mM palmitic acid (2 mM C16:0), or 1 mM palmitic acid plus 1 mM of either stearic (C18:0), oleic (C18:1), linoleic (C18:2), linolenic (C18:3), eicosapentaenoic (C20:5), or docosahexaenoic (C22:6) acid, or 0.5 mM each of eicosapentaenoic and docosahexaenoic acid (C20:5 + C22:6). The two treatments containing 2 mM of saturated fatty acids, 2 mM C16:0 and 1 mM C16:0 plus 1 mM C18:0, increased beta-hydroxybutyrate concentrations in the medium and [1 (14)C]palmitic acid oxidation to acid-soluble products compared with all other treatments. The treatment containing C22:6 increased total cellular triglyceride content and incorporation of [1-(14)C]palmitic acid into cellular triglycerides. The treatments containing C22:6 or C20:5 + C22:6 increased [1-(14)C]palmitic acid metabolism to phospholipids and cholesterol. The presence of C22:6 in the medium decreased metabolism of [2-(14)C]propionic acid either to glucose in the medium or to cellular glycogen. Overall, fatty acids differed in their effects on lipid and glucose metabolism in monolayer cultures of bovine hepatocytes with C22:6 eliciting the most profound changes. PMID- 12906058 TI - Effects of different transition diets on dry matter intake, milk production, and milk composition in dairy cows. AB - The primary objective of this study was to evaluate the effect on dry matter intake (DMI), milk yield, milk composition, body weight (BW), and body condition score (BCS) change of cows offered diets differing in energy density in the last 4 wk of gestation and in the first 8 wk of lactation. Three diets (grass silage:straw, 75:25 on a dry matter basis (SS), grass silage (S), and grass silage + 3 kg concentrate daily (C)) precalving, and two diets (4 kg [LC] or 8 kg [HC] concentrate daily + grass silage ad libitum) postcalving were combined in a 3 x 2 factorial design. Sixty Holstein-Friesian cows entering their second lactation were blocked according to expected calving date and BCS into groups of six and were then allocated at random to the treatments. Individual feeding started 4 wk prior to the expected calving date and measurements were made until the end of the 8th wk of lactation. Mean DMI differed between each of the precalving treatments (7.4, 8.1, and 9.9 kg/d for SS, S, and C, respectively) in the precalving period. The DMI also differed between SS and C for wk 1 to 8 (13.5 and 14.2 kg/d) postcalving. Postcalving, milk (24.2, 26.2, and 28.2 kg/d), fat (933, 1063, and 1171 g/d), and protein (736, 797, and 874 g/d) yields differed between SS, S, and C, respectively. The BCS changes differed between SS and C ( 0.09 and 0.12 of a BCS) in the precalving period and between SS and S compared with C (0.02, 0.06, and -0.26 of a BCS) for wk 1 to 8 postcalving. The BW change differed between SS and S compared with C in both wk 1 to 4 (-0.23, -0.37, and 1.25 kg/d) and wk 1 to 8 (0.18, 0.10, and -0.58 kg/ d) postcalving. The BW and BCS were lower at calving for cows on SS compared with C. The greater amount of concentrate supplement postcalving increased DMI, yields of milk, fat, and protein and decreased BW loss in the first 8 wk of lactation. In conclusion, these results indicate that a greater energy density diet in the final 4 wk of the dry period improves cow production in early lactation. PMID- 12906059 TI - Effects of amino acid nutrition on the responses of dairy cows to milking more frequently with or without injection of growth hormone. AB - The influence of amino acid nutrition on the response to milking more frequently, with or without injection of growth hormone, was examined in eight dairy cows in two 4 x 4 Latin squares with 28-d periods. The four treatments were a diet adequate in amino acids with or without injection of growth hormone and a diet inadequate in amino acids with or without injection of growth hormone. For all four treatments, during the last 14 d of each period, one half of the mammary gland was milked three times a day (3x), while the other half remained on twice daily milking (2x). Both diets were based on grass silage given ad libitum and 4 kg/d of sugar beet pulp together with a supplement containing either fish meal (adequate diet) or feather meal (inadequate diet) as the only protein feeds. The diet containing feather meal is known to be deficient in His, Met, and Lys. On the fish meal diet, the cows responded positively to growth hormone and to milking more frequently and the responses to both treatments were additive. On the feather meal diet, however, even though injection of growth hormone increased the yield of milk protein by around 10%, milking more frequently did not affect milk production. It is concluded that milking more frequently has a weaker effect on the partitioning of amino acid use between body and udder than does growth hormone treatment. PMID- 12906061 TI - Milk production and composition from cows fed high oil or conventional corn at two forage concentrations. AB - Twelve multiparous Holstein cows (63 +/- 24 d in milk) were used in a replicated 4 x 4 Latin square with 28-d periods to evaluate conventional and high oil corn grains when fed at two different forage-to-concentrate ratios. Dietary treatments consisted of conventional or high oil corn supplementing a diet with a 25:25:50 mixture of corn silage: alfalfa: concentrate mix, or a high forage diet with a 30:30:40 mixture of corn silage: alfalfa: concentrate mix. Dry matter intake (28.1, 28.7, 26.9, and 26.2 kg/d for normal diets with conventional and high oil corn, and high forage diets with conventional and high oil corn, respectively) and milk yields (36.8, 37.2, 35.5, and 35.2 kg/d) were similar for conventional and high oil corn diets and were lower with the high forage diet, regardless of corn source. Milk fat concentrations were greater when cows were fed diets containing 60% forage (4.03 vs. 3.88%, for the 60 and 50% forages, respectively), but milk protein concentrations were not affected by forage content. Corn source did not affect milk fat or protein concentrations. Long-chain fatty acid concentrations, unsaturated fatty acid concentrations, and total 18:1 fatty acid concentrations were greater when cows were fed high oil corn but were unaffected by forage content of the diet. Concentrations of transvaccenic acid (0.58, 0.81, 0.62, and 0.69 g/100 g of fatty acids) and cis-9, trans-11 conjugated linoleic acid (0.28, 0.39, 0.32, and 0.33 g/100 g of fatty acids) were greater when cows were fed high oil compared with conventional corn when fed 50% forage but were similar for both corn sources at 60% forage. Total n-3 fatty acids were not affected by corn source or forage content. High forage diets decreased milk production and increased milk fat concentration. Feeding high oil corn increased concentrations of long-chain, unsaturated, transvaccenic, and conjugated linoleic fatty acids in milk; however, production of transvaccenic and conjugated linoleic acids were attenuated by high forage diet. PMID- 12906060 TI - Effect of dietary carbohydrate composition and availability on utilization of ruminal ammonia nitrogen for milk protein synthesis in dairy cows. AB - A trial with four ruminally and duodenally cannulated, late-lactation dairy cows was conducted to investigate the effect of dietary carbohydrate (CHO) composition and availability on ruminal ammonia N utilization and transfer into milk protein. Two diets were fed at 8-h intervals in a crossover design. The diets differed in CHO composition: the ruminally fermentable non-structural carbohydrates (RFSS) diet (barley and molasses) contained a larger proportion of ruminally available CHO in the nonstructural carbohydrate fractions and the ruminally fermentable fiber (RFNDF) diet (corn, beet pulp, and brewer's grains) contained a larger proportion of CHO in ruminally available fiber. Nitrogen-15 was used to label ruminal ammonia N and consequently microbial and milk N. Fermentation acids, enzyme activities, and microbial protein production in the rumen were not affected by diet. Ruminal ammonia concentration was lowered by RFNDF. Ruminal and total tract digestibility of nutrients did not differ between diets except that apparent ruminal degradability of crude protein was lower for RFNDF compared with RFSS. Partitioning of N losses between urine and feces was also not affected by diet. Milk yield and fat and protein content were not affected by treatment. Average concentration of milk urea N was lower for RFNDF than for RFSS. Proportion of milk protein N originating from ruminal microbial N (based on the areas under the 15N-enrichment curves) was higher for RFNDF than for RFSS. Cumulative recovery of 15N in milk protein was 13% higher for RFNDF than for RFSS indicating enhanced transfer of 15N-ammonia into milk protein with the former diet. The results suggested that, compared to diets containing higher levels of ruminally fermentable starch, diets providing higher concentration of ruminally fermentable fiber may enhance transfer of ruminal ammonia and microbial N into milk protein. PMID- 12906062 TI - The effect of corn silage particle size and cottonseed hulls on cows in early lactation. AB - The objective of this study was to evaluate the effects of reducing forage particle length (FPL) and the inclusion of cottonseed hulls (CSH) on intake, digestibility, chewing activity, and milk production of cows in early lactation. Sixteen multiparous cows averaging 17 +/- 3 d in milk and 677 +/- 58 kg BW were assigned to one of four 4 x 4 Latin squares. One square contained ruminally cannulated cows to evaluate effects of treatment on rumen fermentation and function. During each of the 23-d periods, cows were offered one of four total mixed rations that differed in particle length (long or short corn silage) and CSH inclusion rate (0 or 8% DM). Dietary treatments were: long no CSH (LGNH), long with CSH (LGH), short no CSH (SHNH), and short with CSH (SHH). Total physically effective NDF content, measured as percentage of NDF greater than 1.18 mm, was similar across diets, but mean particle length decreased with reducing FPL and inclusion of CSH. Dry matter intake was not significantly affected by FPL but was significantly increased with the inclusion of CSH. Decreasing FPL and the inclusion of CSH significantly increased neutral detergent fiber intake. Total chewing activity expressed as minutes per day was unaffected by FPL and the inclusion of CSH. Both eating and ruminating efficiency expressed as minutes per kilogram of neutral detergent fiber intake increased with increasing FPL and decreased with the inclusion of CSH. Milk production did not differ across treatments, but the inclusion of CSH significantly increased percent and yield of milk protein. Reducing FPL tended to reduce percentage milk fat but not yield. Mean ruminal pH was not affected by FPL but was highest on diets containing CSH, even though no treatment effects were observed on total VFA, acetate, or propionate concentration. These resuits indicate that corn silage FPL is a poor predictor of total chewing time and rumen pH but is useful in understanding factors affecting feeding behavior. In addition, the inclusion of CSH, resulted in increased rumination and mean rumen pH even though effects on VFA concentration were not observed. PMID- 12906063 TI - Effects of milk products fermented by Bifidobacterium longum on blood lipids in rats and healthy adult male volunteers. AB - The effects of milk products fermented by Bifidobacterium longum strain BL1, a probiotic strain, on blood lipids in rats and humans were studied. Rats were fed a cholesterol-enriched experimental diet, supplemented with lyophilized powders of 1) acid milk (control), 2) milk fermented with a mixed culture of ordinary yogurt starters composed of Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus (SL), and 3) bifidobacterium milk fermented with the probiotic B. longum strain BL1, respectively. The bifidobacterium milk feeding brought about significant lowering of the serum concentrations of total cholesterol, low-density lipoprotein cholesterol, and triglycerides, in comparison with the control, while no change in high-density lipoprotein cholesterol concentration was observed. On the other hand, supplementation with SL milk resulted in only slight, nonsignificant decreases in serum lipid concentrations in comparison with the control. In the human study, 32 subjects with serum total cholesterol ranging from 220 to 280 mg/dl were randomly assigned to two treatments: 1) intake of a low-fat drinking yogurt prepared with ordinary yogurt starters composed of S. thermophilus and L. delbrueckii subsp. bulgaricus (P-group) and 2) intake of a low-fat drinking yogurt prepared with the two ordinary yogurt starters plus B. longum strain BL1 (B-group). After intake for 4 wk at 3 x 100 ml/day, reduction of serum total cholesterol was observed in approximately half of the B-group subjects; a particularly significant decrease in serum total cholesterol was found among subjects with moderate hypercholesterolemia (serum total cholesterol > 240 mg/dl). However, the serum lipid concentrations in the P-group subjects were almost stable during the experimental periods. The present results indicate the potential of the probiotic B. longum strain BL1 in serum lipid improvement. PMID- 12906064 TI - Estimation of genetic parameters for concentrations of milk urea nitrogen. AB - The objective of this study was to use field data collected by dairy herd improvement programs to estimate genetic parameters for concentrations of milk urea nitrogen (MUN). Edited data were 36,074 test-day records of MUN and yields of milk, fat, and protein obtained from 6102 cows in Holstein herds in Ontario, Canada. Data were divided into three sets, for the first three lactations. Two analyses were performed on data from each lactation. The first procedure used ANOVA to estimate the significance of the effects of several environmental factors on MUN. Herd-test-day effects had the most significant impact on MUN. Effects of stage of lactation were also important, and MUN levels tended to increase from the time of peak yield until the end of lactation. The second analysis used a random regression model to estimate heritabilities and genetic correlations of MUN and the yield traits. Heritability estimates for MUN in lactations one, two, and three were 0.44, 0.59, and 0.48, respectively. Heritabilities for the yield traits were of a similar magnitude. Little relationship was observed between MUN and yield. Raw phenotypic correlations were all <0.10 (absolute value). Genetic correlations with production traits were close to zero in lactations one and three and only slightly positive in lactation two. The results indicate that selection on MUN is possible, but relationships between MUN and other economically important traits such as metabolic disease and fertility are needed. PMID- 12906066 TI - Modeling lactation curves and estimation of genetic parameters for first lactation test-day records of French Holstein cows. AB - Several functions were used to model the fixed part of the lactation curve and genetic parameters of milk test-day records to estimate using French Holstein data. Parametric curves (Legendre polynomials, Ali-Schaeffer curve, Wilmink curve), fixed classes curves (5-d classes), and regression splines were tested. The latter were appealing because they adjusted the data well, were relatively insensitive to outliers, were flexible, and resulted in smooth curves without requiring the estimation of a large number of parameters. Genetic parameters were estimated with an Average Information REML algorithm where the average information matrix and the first derivatives of the likelihood functions were pooled over 10 samples. This approach made it possible to handle larger data sets. The residual variance was modeled as a quadratic function of days in milk. Quartic Legendre polynomials were used to estimate (co)variances of random effects. The estimates were within the range of most other studies. The greatest genetic variance was in the middle of the lactation while residual and permanent environmental variances mostly decreased during the lactation. The resulting heritability ranged from 0.15 to 0.40. The genetic correlation between the extreme parts of the lactation was 0.35 but genetic correlations were higher than 0.90 for a large part of the lactation. The use of the pooling approach resulted in smaller standard errors for the genetic parameters when compared to those obtained with a single sample. PMID- 12906065 TI - Genotype by environment interaction for yield and somatic cell score with alternative environmental definitions. AB - Differential genetic expression in high and low opportunity Sicilian Holstein Friesian and Brown Swiss herd environments was investigated using endogenous and exogenous variables in a set of three definitions. Results of genetic by environmental interaction were compared using alternative environmental definitions: within herd-year standard deviation for mature equivalent milk yield (HYSD), detectable incidence of normal vs. abnormal (peakless) lactation and herds clustered by causal relationships from high and low frequency use of nutrition, milking, health and animal handling practices. Data for genetic analysis consisted of first-lactation standardized yields of milk, fat and protein, and weighted somatic cell score for 8897 daughters of 825 Holstein Friesian sires and 1143 daughters of 220 Brown Swiss sires. Components of covariance, heritabilities, and genetic correlations were estimated using bivariate and multivariate sire models for average and contrasting environments for each definition. Sire variances for yields were consistently smaller in the low opportunity environments of both breeds. Except for differential incidence of abnormal lactation in Friesian herds, correlated yield response in less privileged environments was 0.41 to 0.81 as much as in high opportunity environments, a substantial loss. Genetic correlations between HYSD environments for yield traits of Friesian were 0.48 to 0.66 but exceeded 0.80 for other definitions. Less correlated response in somatic cell score was also predicted for environments with low use of yield-enhancing practices (0.66 for Friesian and 0.61 for Brown Swiss), which may have resulted from less health care and poorer milking management. Therefore, unfavorable management interactions likely foster unequal gains from selection in contrasting environments defined exogenously or by incidence of peakless lactation. Conversely, greater genetic as well as phenotypic response is expected from additional inputs of nutrition, health care and milking management. PMID- 12906067 TI - Polarization-mode dispersion measurement by an optical time-domain reflectometer with polarimetry assuming backscattering by randomly oriented nonspherical particles. AB - In previous work on polarization-mode dispersion (PMD) measurement with an optical time-domain reflectometer with polarimetry (p-OTDR), scatterers were assumed to be a cloud of small spherical particles in a fiber. We have found that the p-OTDR waveforms were fitted well by modified Mueller matrices, assuming scattering by a cloud of nonspherical particles. We realized a PMD measurement based on the p-OTDR Jones matrix eigenanalysis (JME) method. The measured PMD was consistent with that of JME standard measurement with transmission-type polarimetry. PMID- 12906068 TI - Poled polymer thin-film gratings studied with far-field optical diffraction and second-harmonic near-field microscopy. AB - Electrical poling induces polar ordering of molecules in a grating that has been holographically inscribed on a thin film of polymer with azobenzene side chains. The resulting chi2 grating, seen by second-harmonic-generation (SHG) near-field scanning optical microscopy, can have a periodic structure that is significantly different from the topographical image. The far-field linear and SHG diffration patterns correlate well with the grating structures. Poling of the thin-film grating, which presumably has photodriven nonuniform material properties within each period, leads to the more complex structure of the chi2 grating. PMID- 12906069 TI - Spatial separation of the traveling and evanescent parts of dipole radiation. AB - Electric dipole radiation consists of traveling and evanescent plane waves. When radiation is detected in the far field, only the traveling waves will contribute to the intensity distribution, as the evanescent waves decay exponentially. We propose a method to spatially separate the traveling and evanescent waves before detection. It is shown that when the radiation passes through an interface, evanescent waves can be converted into traveling waves and can subsequently be observed in the far field. Let the radiation be observed under angle theta(t) with the normal. Then there exists an angle theta(ac) such that for 0 < or = theta(t) < theta(ac) all intensity originates in traveling waves, whereas for theta(ac) < theta(t) < pi/2 only evanescent waves contribute. It is shown that with this technique and under the appropriate conditions almost all far-field power can be provided by evanescent waves. PMID- 12906070 TI - Nanotaper for compact mode conversion. AB - We propose and demonstrate an efficient coupler for compact mode conversion between a fiber and a submicrometer waveguide. The coupler is composed of high index-contrast materials and is based on a short taper with a nanometer-sized tip. We show that the micrometer-long silicon-on-insulator-based nanotaper coupler is able to efficiently convert both the mode field profile and the effective index, with a total length as short as 40 microm. We measure an enhancement of the coupling efficiency between an optical fiber and a waveguide by 1 order of magnitude due to the coupler. PMID- 12906071 TI - 90-fs stretched-pulse ytterbium-doped double-clad fiber laser. AB - We report ultrashort, high-energy pulse generation from a side-pumped Yb-doped double-clad fiber laser. The laser is operating in the stretched-pulse regime. It directly generates 4.8-ps, 5-nJ pulses that are extracavity compressed to 90 fs. PMID- 12906072 TI - Tunable all-optical negative multitap microwave filters based on uniform fiber Bragg gratings. AB - We present a novel and simple technique for obtaining transversal filters with negative coefficients by using uniform fiber Bragg gratings. We demonstrate a wide tuning range, good performance, low cost, and easy implementation of multitap filters in an all-optical passive configuration in which negative taps are obtained by use of the transmission of a broadband source through uniform Bragg gratings. PMID- 12906073 TI - Threshold features of a Brillouin Stokes comb generated in a distributed fiber Raman amplifier. AB - We study threshold features of a Brillouin-shifted Stokes comb generated in a distributed fiber Raman amplifier. When the input power of Brillouin pump is linearly increased in high Raman gain, the first Stokes wave grows exponentially at much lower threshold power and then experiences an appreciable power decrease in the vicinity of the Brillouin comb's threshold. This power reduction of the first Brillouin Stokes, which we did not see mentioned in previous reports, was caused by a power transfer to higher-order lines and initiated Brillouin comb generation. Moreover, the effects of Raman pump power and pumping direction on the threshold of a Brillouin comb are investigated. PMID- 12906074 TI - Far-field photothermal microscopy beyond the diffraction limit. AB - The principles of laser photothermal (PT) imaging beyond the diffraction limit are discussed. Algorithms for obtaining information on the sizes and the shapes of nanotargets are analyzed. Spectral PT lifetime imaging is suggested to provide spatial localization of nanotargets overlapping inside the diffraction volume. PMID- 12906075 TI - Direct measurement of the spatial Wigner function with area-integrated detection. AB - We demonstrate experimentally a novel technique for characterizing transverse spatial coherence by using the Wigner distribution function. The method is based on the measurement of interference between a pair of rotated and displaced replicas of the input beam with an area-integrating detector, and it provides an optimal signal-to-noise ratio in regimes when array detectors are not available. We analyze the quantum-optical picture of the presented measurement for single photon signals and discuss possible applications in quantum information processing. PMID- 12906077 TI - Low-coherence interferometric tip-clearance probe. AB - We propose an all-fiber, self-calibrating, economical probe that is capable of near-real-time, single-port, simultaneous blade-to-blade tip-clearance measurements with submillimeter accuracy (typically < 100 microm, absolute) in the first stages of a gas turbine. Our probe relies on the interference between backreflected light from the blade tips during the 1-micros blade passage time and a frequency-shifted reference with variable time delay, making use of a low coherence light source. A single optical fiber of arbitrary length connects the self-contained optics and electronics to the turbine. PMID- 12906078 TI - Inhibition of modulation instability in lasers by noise. AB - It is shown that additive noise can inhibit modulation instability in laser equations of motion. A related self-starting condition for pulsation is obtained by employing a fluctuation--dissipation relation between noise and losses and a statistical mechanics approach. Entropy considerations are shown to play a crucial role. The quantum limit for self-starting is estimated. PMID- 12906076 TI - Ultrahigh light transmission through a C-shaped nanoaperture. AB - Optical resolution beyond the diffraction limit can be achieved by use of a metallic nanoaperture in a near-field optical system. Conventional nanoapertures have very low power throughput. Using a numerical finite-difference time domain method, we discovered a unique C-shaped aperture that provides approximately 3 orders of magnitude more power throughput than a conventional square aperture with a similar near-field spot size of approximately 0.1 lambda. Microwave experiments at 6 GHz quantitatively confirmed the simulated transmission enhancement. The high transmission of the C-aperture--or one of the related shapes--is linked to both a propagation mode in the aperture and local surface plasmons. PMID- 12906079 TI - Optical heterodyne surface-plasmon resonance biosensor. AB - A novel optical heterodyne surface-plasmon resonance (SPR) biosensor with a Zeeman laser is proposed. Two surface plasma waves are excited by two correlated p-polarized waves in a SPR device of the Kretschmann configuration. Two reflected p waves are optically heterodyned such that the magnitude of the heterodyned signal is proportional to the multiplication of two attenuated reflected p waves. Then the detection sensitivity and the dynamic range based on this amplitude sensitive method are enhanced. In the experiment, the kinetics between mouse immunoglobulin G (IgG) and rabbit antimouse IgG is obtained from sensograms of various concentrations of antimouse IgG. A detection sensitivity of 0.2 nM was achieved. In addition, a concentration of 5 ng/ml of protein G interacting with mouse IgG was measured successfully. PMID- 12906080 TI - Differential spectral interferometry: an imaging technique for biomedical applications. AB - Differential spectral interferometry (DSI), a novel method of biomedical imaging that combines the high dynamic range of optical coherence tomography (OCT) with inherently parallel low-bandwidth image acquisition of spectral interferometry (SI), is described. DSI efficiently removes the deleterious dc background inherent in SI measurements while maintaining the parallel nature of SI. We demonstrate DSI on both synthetic and biological samples. Because DSI preserves the low-bandwidth, parallel nature of SI, it is competitive with OCT for biomedical applications in terms of image quality and acquisition rate. PMID- 12906081 TI - Optical trapping microfabrication with electrophoretically delivered particles inside glass capillaries. AB - We have developed a new technique for rapid microfabrication that uses electrophoretically delivered particles and an optical trap. The material particles, micrometer- and nanometer-sized polystyrene beads in aqueous solution, are continuously delivered to an optical trap by means of the electrophoretic effect inside glass capillaries or similar microstructures. The optical trap is used to manipulate and deposit the polystyrene beads onto a substrate. The continuous, on-demand delivery of particles allows for microfabrication in two and three dimensions with high speed and high efficiency and without material waste. This new technique has many potential applications in microelectronics and biotechnology. PMID- 12906082 TI - Three-dimensional electric field visualization utilizing electric-field-induced second-harmonic generation in nematic liquid crystals. AB - An electric-field-induced second-harmonic-generation signal in a nematic liquid crystal is used to map the electric field in an integrated-circuit-like sample. Since the electric-field-induced second-harmonic-generation signal intensity exhibits a strong dependence on the polarization of the incident laser beam, both the amplitude and the orientation of the electric field vectors can be measured. Combined with scanning second-harmonic-generation microscopy, three-dimensional electric field distribution can be easily visualized with high spatial resolution of the order of 1 microm. PMID- 12906083 TI - K5Nd(MoO4)4: a self-tunable laser crystal. AB - We report for the first time to our knowledge an experimental demonstration of wavelength self-tuning in a K5Nd(MoO4)4 broadband-emitting laser crystal (KNM), as well as a theoretical treatment of the system based on its birefringence properties. The self-frequency-tuning of the laser along the full spectral range of the KNM crystal was obtained by rotating a birefringent gain plate in its own plane. The gain plate was placed inside a resonator at Brewster's angle. The tuning characteristics of the spectral filter were obtained by use of the Jones vector formalism. The calculated wavelength-selective tuning precisely matches the experimental observations. PMID- 12906084 TI - Efficient 340-nm light generation by a ridge-type waveguide in a first-order periodically poled MgO:LiNbO3. AB - Quasi-phase-matched (QPM) UV second-harmonic generation (SHG) in a periodically poled MgO:LiNbO3 waveguide is presented. A ridge-type waveguide with high nonlinearity and strong resistance to photorefractive damage was achieved by use of an ultraprecision machining technique. By use of this waveguide in 1.4-microm periodically poled MgO:LiNbO3, a first-order QPM SHG device for 340-nm UV radiation was demonstrated. In a single-pass configuration, continuous-wave 22.4 mW UV light was generated for a fundamental power of 81 mW, corresponding to a normalized conversion efficiency of 340%/W. PMID- 12906085 TI - Nonlinear optical switching matrix. AB - We show that nonlinear directional couplers with variable coupling coefficients can be used to compose a special kind of switching matrix. The switching matrix can be controlled by the signal itself or by a strong pump. Light routing is realized by adjustment of the power of the signal or the pump. Because light routing is based on the optical Kerr effect, ultrafast routing is possible. PMID- 12906086 TI - Asymptotic probability density of nonlinear phase noise. AB - The asymptotic probability density of nonlinear phase noise, often called the Gordon-Mollenauer effect, is derived analytically when the number of fiber spans is large. Nonlinear phase noise is the summation of infinitely many independently distributed noncentral chi2 random variables with two degrees of freedom. The mean and the standard deviation of those random variables are both proportional to the square of the reciprocal of all odd natural numbers. Nonlinear phase noise can also be accurately modeled as the summation of a noncentral chi2 random variable with two degrees of freedom and a Gaussian random variable. PMID- 12906087 TI - Continuous-wave, high-power, Raman continuum generation in holey fibers. AB - The possibility of using low pump power for cw Raman continuum generation is demonstrated by optimization of the pump peak power and by accounting for the loss-related reduction of the effective length of Raman interaction in holey fibers. A 3.8-W, 324-nm-wide cw Raman continuum with a spectral power density higher than 10 mW/nm is generated in a completely fiber-integrated, single-mode format. PMID- 12906088 TI - Compact confocal readout system for three-dimensional memories using a laser feedback semiconductor laser. AB - We present a compact confocal readout system for three-dimensional optical memories that uses the thresholding property of a semiconductor laser for feedback light. The system has higher axial resolution than a conventional confocal system with a pinhole. We demonstrate readout results for data recorded in two recording layers with the developed system. PMID- 12906089 TI - Complex modulation with a twisted-nematic liquid-crystal spatial light modulator: double-pixel approach. AB - I modify the double-phase holographic code to implement complex modulation with a transmission twisted-nematic liquid-crystal display. This device is employed in the mostly phase configuration, for which the phase modulation is coupled with a nonconstant amplitude modulation. The modified double-phase code implements arbitrary complex modulation employing the constrained complex modulation of the display. PMID- 12906090 TI - Practical all-fiber source of high-power, 120-fs pulses at 1 micrometre. AB - Amplification of femtosecond pulses at 1.03 micrometre in a standard Yb-doped single-mode fiber is reported. A pulse energy of 8 nJ and an average power of 400 mW are obtained, limited by available pump power. To our knowledge these are the highest pulse energy and average power obtained from an integrated, single-mode fiber amplifier. After dechirping, 120-fs, 6-nJ pulses are obtained. A practical fiber-based source with performance comparable with that of a bulk solid-state laser is thus demonstrated, and scaling to substantially higher powers will be possible. PMID- 12906091 TI - Generation of 50-fs, 5-nJ pulses at 1.03 micrometre from a wave-breaking-free fiber laser. AB - We report the generation of 6-nJ chirped pulses from a mode-locked Yb fiber laser at 1.03 micrometre. A linear anomalous-dispersion segment suppresses wave breaking effects of solitonlike pulse shaping at high energies. The dechirped pulse duration is 50 fs, and the energy is 5 nJ. This laser produces twice the pulse energy and average power, and approximately five times the peak power, of the previous best mode-locked fiber laser. It is to our knowledge the first fiber laser that directly offers performance similar to that of solid-state lasers such as Ti:sapphire. PMID- 12906092 TI - 420-MHz Cr:forsterite femtosecond ring laser and continuum generation in the 1-2 micrometre range. AB - We demonstrate a chromium-doped forsterite femtosecond ring laser that generates 30-fs pulses at a 420-MHz repetition rate with nearly 500 mW of average power. The compact solid-state design and broad spectral output make this laser attractive for telecommunications applications in the 1.3-1.5-micrometre region. Additional spectral broadening of the laser output in highly nonlinear optical fiber leads to octave-spanning spectra ranging from 1.06 to 2.17 micrometre. The octave is reached at a level of 18 dB below the peak. The underlying optical frequency comb can be linked to existing optical frequency standards. PMID- 12906093 TI - Dispersion-managed semiconductor mode-locked ring laser. AB - A novel breathing-mode external sigma-ring-cavity semiconductor mode-locked laser is developed. Intracavity pulse compression and stretching produce linearly chirped pulses with an asymmetric exponential temporal profile. External dispersion compensation reduces the pulse duration to 274 fs (within 10% of the bandwidth limit). PMID- 12906094 TI - Nd:glass diode-pumped regenerative amplifier, multimillijoule short-pulse chirped pulse-amplifier laser. AB - We have built a diode-pumped Nd:glass regenerative amplifier that is able to produce energies up to 20 mJ within a 470-fs pulse duration at a 1-Hz repetition rate. We obtained this amplifier by using specific intracavity components such as a phase mirror and a birefringent filter to generate a large spatial mode and a large spectral width. PMID- 12906095 TI - Optical phase locking among femtosecond subharmonic pulses. AB - We stabilized the relative carrier-envelope phase slip among the pump pulse and its subharmonic signal and idler pulses in a femtosecond optical parametric oscillator, resulting in long-term phasecoherence among the pulses. The stabilized beat signal corresponding to the relative carrier-envelope phase slip among subharmonic pulses had an accumulated phase error of 0.24 rad in the 1-mHz 1-MHz region. The fluctuation of the beat frequency measured by a 1-s-averaged counter was less than 1 mHz in a 1480-s measurement. PMID- 12906096 TI - The suppressive effect of bifidobacteria on Bacteroides vulgatus, a putative pathogenic microbe in inflammatory bowel disease. AB - Bacteroides, a predominant commensal bacteria in the gut, are thought to be responsible for the development of inflammatory bowel disease (IBD). In the present study, we examined whether or not bifidobacteria suppress B. vulgatus, a representative pathogenic Bacteroides species, in both the coculture system and the gnotobiotic murine model. As a result, Bifidobacterium infantis 1222 highly inhibited the growth of B. vulgatus in the coculture and also significantly suppressed the systemic antibody response raised by B. vulgatus colonizing the gut in gnotobiotic mice. Colonization of the mice by B. vulgatus increased the number of Peyer's patch (PP) cells bearing PNA (peanut agglutinin)+/anti-kappa+ phenotype, which represents plasma cell-like B cells. Moreover, treatment of those B. vulgatus-implanted mice with B. infantis 1222 abrogated such increase in the number of PNA+/anti-kappa+ cells. These results thus suggested that B. infantis 1222 protected the gut epithelial layer including the PP from being invaded by Bacteroides, thereby suppressing the systemic antibody response raised by Bacteroides. PMID- 12906098 TI - Degradation process of Mycobacterium leprae cells in infected tissue examined by the freeze-substitution method in electron microscopy. AB - Mycobacterium leprae cells (strain Thai-53) harvested from infected mouse foot pads were examined by electron microscopy using the freeze-substitution technique. The population of M. leprae cells from the infected tissue consisted of a large number of degraded cells and a few normal cells. These thin sectioned cell profiles could be categorized into four groups depending on the alteration of the membrane structures, and the degradation process is considered to occur in stages, namely from stages 1 to 3. These are the normal cells with an asymmetrical membrane, a seemingly normal cell but with a symmetrical membrane (stage 1), a cell possessing contracted and highly concentrated cytoplasm with a membrane (stage 2), and a cell that has lost its membrane (stage 3). The peptidoglycan layer was found to remain intact in these cell groups. PMID- 12906097 TI - Possible relationship of PFGE patterns of Moraxella catarrhalis between hospital- and community-acquired respiratory infections in a community hospital. AB - We describe a prospective study of molecular analysis of Moraxella catarrhalis isolated from a community hospital. Our study was designed to investigate the possible relationship of pulsed-field gel electrophoresis (PFGE) patterns of M. catarrhalis between hospital- and community-acquired respiratory infections. A nosocomial outbreak of M. catarrhalis was observed between September 2000 and September 2001. During the study period, 40 strains of M. catarrhalis were isolated from a total of 32 patients with respiratory infections (26 strains from 18 inpatients, and 14 strains from 14 outpatients). We compared the PFGE patterns in 40 strains of M. catarrhalis isolated from the respiratory tract of the study patients. The genomic types of M. catarrhalis were classified into three PFGE patterns (A, B, and C). Interestingly, the nosocomial outbreak of M. catarrhalis included two patterns (A and B). Of the three patterns, two patterns (A and B) were found in both inpatients and outpatients. More interestingly, two subtypes of pattern B (B1 and B4) were simultaneously found in both inpatients and outpatients. Our results indicated that PFGE with SmaI chromosomal digestion is a suitable technique to establish the inter-strain genetic relatedness of M. catarrhalis, and suggested that the outbreak of M. catarrhalis occasionally included miscellaneous PFGE patterns. The results also showed that PFGE patterns of M. catarrhalis isolates were similar between hospital- and community-acquired respiratory infections. Analysis of the subtypes suggested that there might be some association between hospital- and community-acquired respiratory infections caused by M. catarrhalis. PMID- 12906099 TI - Disruption of the human pathogenic yeast Candida albicans catalase gene decreases survival in mouse-model infection and elevates susceptibility to higher temperature and to detergents. AB - Catalase-deficient strains of the human pathogenic yeast Candida albicans were constructed using the URA-blaster method. The disruptant was viable and grew normally in an ordinary culture condition, but became extremely sensitive to treatment with hydrogen peroxide. No catalase activity was observed in a catalase (CCT)-gene-disrupted strain, 1F5-4-1, suggesting that there were no other catalase or catalase-like enzymes in this yeast. The disruptant was shown to be sensitive to higher temperature and to low concentrations of SDS, NP-40, or Triton X-100. After a wild-type CCT gene was reintroduced into the disruptant, catalase activity was restored and the strain became moderately sensitive to treatment with hydrogen peroxide. However, neither the temperature sensitivity nor the susceptibility to SDS observed in the disruptant was restored in the CCT reintroduced strain. A model infection experiment using wild-type and dCCT strains showed that the disruptants disappeared more rapidly than the wild-type strain in mouse liver, lung, and spleen. These results suggest that the catalase plays a significant role in survival in the host immune system and thus leads this organism to establish infection in the host. PMID- 12906100 TI - The effect of Lactobacillus rhamnosus on enterohemorrhagic Escherichia coli infection of human intestinal cells in vitro. AB - There are many examples of probiotic effects of various lactic bacteria on enteropathogens. In this study, Lactobacillus strains (L. rhamnosus, L. gasseri, L. casei and L. plantarum) were tested in an in vitro model of enterohemorrhagic Escherichia coli (EHEC) infection of a human colon epithelial cell line, C2BBe1. While the adhesion and colonization of EHEC was not affected by any of the lactobacillus strains tested, the internalization of EHEC into the cell line was markedly suppressed by L. rhamnosus, though not by others. Concerning the possible mechanisms, the viabilities of EHEC and host cell were not affected by the presence of L. rhamnosus. Simple competitions at certain receptors were unlikely because the suppressive effect on EHEC internalization was strictly dependent on viable L. rhamnosus and could not be observed with the conditioned medium or killed L. rhamnosus. The fact that L. rhamnosus showed outstanding potential for adhering to the colon epithelial cell line, compared with other strains, suggested that an avid interaction between L. rhamnosus and the host cell might be modulating intra-cellular events responsible for the internalization of EHEC. PMID- 12906101 TI - Cloning and characterization of the fur gene from Moraxella bovis. AB - A homologue of the ferric uptake regulator gene (fur) was isolated from Moraxella bovis by degenerate polymerase chain reaction and cloning. Fur protein of M. bovis exhibited 72.1% amino acid identity with Acinetobacter calcoaceticus Fur. Western blot analysis showed a decrease of Fur expression in response to sufficient-iron conditions compared with deficient-iron conditions. An electrophoretic mobility-shift assay indicated that Fur protein binds to DNA fragments containing a putative Fur-box derived from the upstream region of the M. bovis fur gene. Fur of M. bovis may regulate the expression of iron transport systems in response to iron limitation in the environment. PMID- 12906102 TI - Gene cloning and characterization of VcrM, a Na+-coupled multidrug efflux pump, from Vibrio cholerae non-O1. AB - We cloned a DNA fragment responsible for drug resistance from chromosome of Vibrio cholerae non-O1. Nucleotide sequence analysis of this fragment revealed the presence of a single open reading frame encoding a protein consisting of 445 amino acid residues. We designated the gene as vcrM. Hydropathy analysis of the deduced amino acid sequence of VcrM suggests the presence of 12 trans-membrane segments. A dendrogram showed that VcrM is a member of the DinF-subfamily within the MATE family of multidrug efflux pumps. Expression of the cloned vcrM gene in drug-hypersensitive Escherichia coli KAM32 cells made them resistant to acriflavine, 4', 6-diamidino-2-phenylindole, Hoechst 33342, rhodamine 6G, tetraphenylphosphonium chloride (TPPCl) and ethidium bromide. Efflux of acriflavine due to VcrM was dependent on Na+ or Li+. Moreover, Na+ efflux was observed with VcrM when TPPCl was added to Na+-loaded cells. Therefore, we conclude that VcrM is a Na+/drug antiporter-type multidrug efflux pump. PMID- 12906103 TI - Hepatitis C virus core protein selectively inhibits synthesis and accumulation of p21/Waf1 and certain nuclear proteins. AB - By using a vaccinia virus-T7 expression system, possible effects of hepatitis C virus (HCV) core protein on synthesis and accumulation of host cellular proteins transiently expressed in cultured cells were analyzed. Immunoblot and immunofluorescence analyses revealed that synthesis and accumulation of certain nuclear proteins, such as p21/Waf1, p53, proliferating cell nuclear antigen and c Fos, were strongly inhibited by HCV core protein. On the other hand, synthesis and accumulation of cytoplasmic proteins, such as 2'-5'-oligoadenylate synthetase (2'-5'-OAS), RNase L and MEK1, were barely affected by HCV core protein. Northern blot analysis showed that the degrees of mRNA expression for those proteins did not differ between HCV core protein-expressing cells and the control, suggesting that the inhibition occurred at the post-transcription level. Pulse-labeling analysis suggested that HCV core protein strongly inhibited synthesis of p21/Waf1 at the translation level. Once being accumulated in the nucleus, p21/Waf1 stability was not significantly affected by HCV core protein. Mutants of HCV core protein C-terminally deleted by 18 or 41 amino acids (aa), which were localized almost exclusively in the nucleus, lost their ability to inhibit synthesis/accumulation of p21/Waf1 whereas another mutant C-terminally deleted by 8 aa still maintained the same properties (subcellular localization and the inhibitory effect) as the full-length HCV core protein of 191 aa. Taken together, our present results suggest that expression of HCV core protein in the cytoplasm selectively inhibits synthesis of p21/Waf1 and some other nuclear proteins at the translation level. PMID- 12906104 TI - Discrimination of West Nile virus and Japanese encephalitis virus strains using RT-PCR RFLP analysis. AB - West Nile (WN) virus is a mosquito-borne flavivirus that induces lethal encephalitis in humans and horses. Since an outbreak of WN encephalitis in humans and horses occurred in New York City in late August 1999, the possibility exists that WN virus will invade regions that have close links with the United States, such as Japan. We developed a genetic diagnostic method that discriminates between strains of WN virus and Japanese encephalitis (JE) virus. The method involves RT-PCR restriction fragment length polymorphism (RFLP) analysis with a RT-PCR primer set, a nested PCR primer set, and a restriction enzyme. We detected WN and JE viruses in experimentally infected animal brain, spleen, and serum samples. Our method is useful in distinguishing WN viruses from the endemic background of JE viruses, and in discriminating the highly virulent WN strain, which was isolated in New York in 1999, from other WN virus strains. PMID- 12906105 TI - High molecular weight vimentin complex is formed after proteolytic digestion of vimentin by caspase-3: detection by sera of patients with interstitial pneumonia. AB - In a previous study, we demonstrated anti-vimentin antibodies in sera of patients with interstitial pneumonia. We hypothesized that antibodies in sera might detect vimentin fragments formed during the process of apoptosis. To prove this, recombinant human vimentin was digested by recombinant human caspase-3 or caspase 8. Then, Western blotting using several commercially available antibodies against human vimentin or patients' sera which had anti-vimentin autoantibodies, was performed. As a result, after recombinant human vimentin was digested by caspase 3 or caspase-8, several vimentin fragments were formed and detected by 2 kinds of monoclonal anti-vimentin antibodies (clone 3B4 and clone V9) as well as by polyclonal sheep anti-human vimentin antibody. It was demonstrated that high molecular weight vimentin was formed after the digestion of vimentin by caspase 3, which was only detected by patients' sera. The high molecular weight vimentin was not formed after digestion of vimentin by caspase-8. Our present results show that high molecular weight vimentin was formed after the digestion of vimentin by caspase-3. In addition, it is suggested that this high molecular weight vimentin acted as an autoantigen to form anti-vimentin autoantibody in vivo. PMID- 12906106 TI - Overcoming tolerance in hepatitis B virus transgenic mice: a possible involvement of regulatory T cells. AB - The hepatitis B virus (HBV) transgenic mouse (Tg) 50-4 strain is immunologically tolerant to HBV antigens. Various vaccination strategies have been attempted but failed to break the tolerance in the mouse. Although the tolerance to HBV antigen is maintained, this mouse strain develops spontaneous liver disease beginning at the age of about 3 months. We attempted to induce immune responses to HBV surface antigen (HBsAg) in the Tg by immunization with recombinant vaccinia virus expressing HBsAg (vvHBV), and observed different immunological responsiveness between 2-month-old and 5-month-old Tg. In contrast to the unbreakable tolerance reported previously, we could induce both the cytotoxic T lymphocyte (CTL) and the antibody response against HBsAg by the vvHBV immunization. The cytokine expression pattern indicated that T helper 1 type immune response was induced. However, interestingly, these immune responses were observed only in the 5-month old Tg, but not in the 2-month-old Tg. Furthermore, CD4+ T cells from 2-month-old mice, but not those from 5-month-old mice, inhibited CTL response to HBV antigen when adoptively transferred to C57BL/6. These results suggest the possible involvement of regulatory T cell function in the HBV Tg for maintaining tolerance. This study would contribute to a better understanding of immune status of the HBV Tg as a model of human chronic hepatitis and to the search for new therapeutic targets for chronic viral infections. PMID- 12906107 TI - Proinflammatory cytokines in bovine colostrum potentiate the mitogenic response of peripheral blood mononuclear cells from newborn calves through IL-2 and CD25 expression. AB - Abstract: Bovine colostrum contains high concentrations of cytokines, and colostral cytokines are considered to be an important factor in stimulation of maturation of the immune system in newborns. In this study, 5 proinflammatory cytokines (IL-1beta, IL-6, TNF-alpha, IFN-gamma and IL-1 receptor antagonist, IL 1ra) present in colostrum were tested for their potential to enhance mitogenic response and to elicit expression of IL-2 mRNA and CD25 in peripheral blood mononuclear cells (PBMC) from newborn calves before being fed colostrum. PBMC were pretreated with each recombinant bovine cytokine for 2 hr before stimulation with concanavalin A (ConA). Pretreatment of PBMC from newborn calves with IL 1beta, TNF-alpha or IFN-gamma significantly enhanced the ConA response, whereas IL-1ra inhibited the response. The degree of enhancement or inhibition of mitogenic response by these cytokines was more pronounced in PBMC from newborn calves than in those from adult cows. Although IL-2 mRNA expression in ConA stimulated PBMC from newborn calves was weaker than that in those from adult cows of ConA-stimulated controls, the expression levels became comparable after pretreatment with IL-1beta, TNF-alpha or IFN-gamma. The CD25 expression in PBMC from newborn calves was also enhanced by pretreatment with IL-1beta, TNF-beta and IFN-gamma. These results suggest that pretreatment of neonatal PBMC with IL 1beta, TNF-alpha or IFN-gamma promotes mitogenic response to ConA through up regulating the production of IL-2 and the expression of the mature IL-2 receptor. PMID- 12906108 TI - Role of microfilaments and microtubules in the invasion of INT-407 cells by Campylobacter jejuni. AB - The internalization mechanisms triggered by Campylobacter jejuni were studied by invasion assays conducted with different inhibitors that act on the cytoskeleton structure of eukaryotic cells. The depolymerization of microfilaments by cytochalasin-D and that of microtubules by colchicines and nocodazole inhibited the uptake of C. jejuni into INT-407 cells in a dose-dependent manner. The inhibitory effect of microfilament depolymerization on C. jejuni internalization was more pronounced than that of microtubule depolymerization. By immunofluorescence microscopic observations, it was demonstrated that both microfilaments and microtubules were localized in INT-407 cells after C. jejuni infection. These data suggest that the internalization mechanism triggered by C. jejuni is associated with the combined effect of microfilaments and microtubules of host cells. PMID- 12906109 TI - DNA base alignment and taxonomic study of genus Malassezia based upon partial sequences of mitochondrial large subunit ribosomal RNA gene. AB - The sequences of the large subunit of mitochondrial ribosomal RNA (LsmtrRNA) gene of Malassezia species were analysed. The sequences of the seven species of Malassezia are well separated in each species. Therefore the LsmtrRNA gene is thought to be one of the gene targets for species identification in the genus Malassezia. The dendrogram obtained from this gene supports the previous study of Malassezia species based upon the chromosomal genes. This is the first report of taxonomic analysis of Malassezia species based upon the mitochondrial gene. PMID- 12906110 TI - Gene silencing by expression of hairpin RNA in Lotus japonicus roots and root nodules. AB - We investigated the efficacy of self-complementary hairpin RNA (hpRNA) expression to induce RNA silencing in the roots and nodules of model legume Lotus japonicus, using hairy root transformation mediated by Agrobacterium rhizogenes. Transgenic lines that express beta-glucuronidase (GUS) by constitutive or nodule-specific promoters were supertransformed by infection of A. rhizogenes harboring constructs for the expression of hpRNAs with sequences complementary to the GUS coding region. GUS activity in more than 60% of the hairy roots was decreased or silenced almost completely. Silencing of the GUS gene was also observed in symbiotic nodules formed on hairy roots in both early and late stages of nodule organogenesis. These results indicate that transient RNA silencing by hairy root transformation provides a powerful tool for loss-of-function analyses of genes that function in roots and root nodules. PMID- 12906111 TI - Targeted activation tagging of the Arabidopsis NBS-LRR gene, ADR1, conveys resistance to virulent pathogens. AB - A transgenic Arabidopsis line containing a chimeric PR-1::luciferase (LUC) reporter gene was subjected to mutagenesis with activation tags. Screening of lines via high-throughput LUC imaging identified a number of dominant Arabidopsis mutants that exhibited enhanced PR-1 gene expression. Here, we report the characterization of one of these mutants, designated activated disease resistance (adr) 1. This line showed constitutive expression of a number of key defense marker genes and accumulated salicylic acid but not ethylene or jasmonic acid. Furthermore, adr1 plants exhibited resistance against the biotrophic pathogens Peronospora parasitica and Erysiphe cichoracearum but not the necrotrophic fungus Botrytis cinerea. Analysis of a series of adr1 double mutants suggested that adr1 mediated resistance against P. parasitica was salicylic acid (SA)-dependent, while resistance against E. cichoracearum was both SA-dependent and partially NPR1-dependent. The ADR1 gene encoded a protein possessing a number of key features, including homology to subdomains of protein kinases, a nucleotide binding domain, and leucine-rich repeats. The controlled, transient expression of ADR1 conveyed striking disease resistance in the absence of yield penalty, highlighting the potential utility of this gene in crop protection. PMID- 12906112 TI - Tobacco mosaic virus induced alterations in the gene expression profile of Arabidopsis thaliana. AB - In this study, mRNA profiles generated from cDNA microarrays were used to identify gene expression changes in Arabidopsis thaliana ecotype Shahdara infected with Tobacco mosaic virus (TMV). Shahdara is a susceptible TMV host, permitting rapid accumulations of virus in both inoculated and systemic tissues, accompanied by defined disease symptoms that include stunting, necrosis, and leaf curling. Gene expression profiles were monitored in whole tissues of inoculated leaves at four days postinoculation (dpi) and in systemically infected leaves at 14 dpi. Microarrays contained cDNAs representing between 8,000 and 10,000 Arabidopsis genes. Expression analysis identified 68 genes that displayed significant and consistent changes in expression levels, either up or down, in either TMV inoculated or systemically infected tissues, or both. Identified TMV responsive genes encode a diverse array of functional proteins that include transcription factors, antioxidants, metabolic enzymes, and transporters. Thus, the TMV infection process has a significant impact on a wide array of cellular processes that likely reflect the biochemical and physiological changes involved in the development of this disease syndrome. PMID- 12906113 TI - The ABC transporter MgAtr4 is a virulence factor of Mycosphaerella graminicola that affects colonization of substomatal cavities in wheat leaves. AB - The role in virulence of the ATP-binding cassette (ABC) transporters MgAtr1, MgAtr2, MgAtr3, MgAtr4, and MgAtr5 from Mycosphaerella graminicola was analyzed by gene disruption or replacement on seedlings of the susceptible wheat cultivar Obelisk. Disruption strains of MgAtr1 and MgAtr2 and replacement strains of MgAtr3 and MgAtr5 displayed the same phenotype as control strains, while virulence of the MgAtr4 disruption strains was significantly reduced. This reduction in virulence was independent of the wheat cultivar used. Histopathological analysis of the infection process revealed that MgAtr4 disruption strains colonize substomatal cavities less efficiently and display reduced intercellular growth in the apoplast of wheat leaves. In vitro growth experiments in different media showed no fitness penalty associated with the disruption of MgAtr4. Expression analysis demonstrated that transcripts of the constitutively expressed gene CYP51 encoding the fungal-specific cytochrome P450 sterol 14alpha-demethylase from M. graminicola were not detectable in interaction RNA from wheat infected with MgAtr4 disruption strains, thus confirming the reduced intercellular growth of these strains. The results indicate that MgAtr4 is a virulence factor of M. graminicola during pathogenesis on wheat and may function in protection against fungitoxic compounds present around the substomatal cavities of wheat leaves. MgAtr4 is the first virulence factor cloned from this important plant pathogen. PMID- 12906114 TI - Aphid-induced defense responses in Mi-1-mediated compatible and incompatible tomato interactions. AB - The tomato Mi-1 gene confers resistance to three species of root-knot nematode and potato aphid. We studied changes in expression of jasmonic acid (JA)- and salicylic acid (SA)-dependent defense genes in response to potato and green peach aphids. We determined changes in three PR proteins, lipoxygenase and proteinase inhibitors I and II transcripts, locally and systemically in both compatible and incompatible interactions in tomato. Transcripts for PR-1 were detected earlier and accumulated to higher levels in the incompatible than in the compatible potato aphid/tomato interactions. The transcript profiles of the other genes were similar in compatible compared with incompatible interactions. Pin1 and Pin2 RNAs were detected early and transiently in both compatible and incompatible interactions. In tomato plants containing Mi-1, systemic expression of PR-1 and GluB was detected in both compatible and incompatible interactions at 48 h after infestations with either aphid. These results suggest that aphid feeding involves both SA and JA/ethylene plant defense signaling pathways and that Mi-1-mediated resistance might involve a SA-dependent signaling pathway. Potato aphid feeding generated reactive oxygen species in both compatible and incompatible interactions. However, a hypersensitive response was absent in the Mi-1-mediated resistance response to potato aphids. Reciprocal grafting experiments revealed that resistance is cell autonomous, and local expression of Mi-1 is required for Mi-1-mediated resistance against the potato aphid. PMID- 12906115 TI - The Sinorhizobium meliloti glycine betaine biosynthetic genes (betlCBA) are induced by choline and highly expressed in bacteroids. AB - The symbiotic soil bacterium Sinorhizobium meliloti has the capacity to synthesize the osmoprotectant glycine betaine from choline-O-sulfate and choline. This pathway is encoded by the betICBA locus, which comprises a regulatory gene, betI, and three structural genes, betC (choline sulfatase), betB (betaine aldehyde dehydrogenase), and betA (choline dehydrogenase). Here, we report that betICBA genes constitute a single operon, despite the existence of intergenic regions containing mosaic elements between betI and betC, and betB and betA. The regulation of the bet operon was investigated by using transcriptional lacZ (beta galactosidase) fusions and has revealed a strong induction by choline at concentrations as low as 25 microM and to a lesser extent by choline-O-sulfate and acetylcholine but not by osmotic stress or oxygen. BetI is a repressor of the bet transcription in the absence of choline, and a nucleotide sequence of dyad symmetry upstream of betI was identified as a putative betI box. Measurements of intracellular pools of choline, well correlated with beta-galactosidase activities, strongly suggested that BetI senses the endogenous choline pool that modulates the intensity of BetI repression. In contrast to Escherichia coli, BetI did not repress choline transport. During symbiosis with Medicago sativa, S. meliloti bet gene expression was observed within the infection threads, in young and in mature nodules. The existence of free choline in nodule cytosol, peribacteroid space, and bacteroids was demonstrated, and the data suggest that bet regulation in planta is mediated by BetI repression, as in free-living cells. Neither Nod nor Fix phenotypes were significantly impaired in a betI::omega mutant, indicating that glycine betaine biosynthesis from choline is not crucial for nodulation and nitrogen fixation. PMID- 12906116 TI - The parasitome of the phytonematode Heterodera glycines. AB - Parasitism genes expressed in the esophageal gland cells of phytonematodes encode secretions that control the complex process of plant parasitism. In the soybean cyst nematode, Heterodera glycines, the parasitome, i.e., the secreted products of parasitism genes, facilitate nematode migration in soybean roots and mediate the modification of root cells into elaborate feeding cells required to support the growth and development of the nematode. With very few exceptions, the identities of these secretions are unknown, and the mechanisms of cyst nematode parasitism, therefore, remain obscure. The most direct and efficient approach for cloning parasitism genes and rapidly advancing our understanding of the molecular interactions during nematode parasitism of plants is to create gland cell specific cDNA libraries using cytoplasm microaspirated from the esophageal gland cells of various parasitic stages. By combining expressed sequence tag analysis of a gland cell cDNA library with high throughput in situ expression localization of clones encoding secretory proteins, we obtained the first comprehensive parasitome profile for a parasitic nematode. We identified 51 new H. glycines gland-expressed candidate parasitism genes, of which 38 genes constitute completely novel sequences. Individual parasitome members showed distinct gland cell expression patterns throughout the parasitic cycle. The parasitome complexity discovered paints a more elaborate picture of host cellular events under specific control by the nematode parasite than previously hypothesized. PMID- 12906117 TI - Identification of extracytoplasmic proteins in Bradyrhizobium japonicum using phage display. AB - A novel gene bank of Bradyrhizobium japonicum USDA110spc4 was constructed using pG3DSS, a phagemid vector designed for detecting genes encoding secreted proteins. In this phagemid, the phage protein III lacks its indigenous signal peptide required for protein secretion, thus recombinant fusion proteins are displayed on the phage surface only if a functional signal peptide is provided by an inserted DNA fragment. In addition, the N-terminal half of protein III has been replaced by a short linker region (the E-tag) that is recognized by a monoclonal antibody, which enables isolation of phages displaying a fusion protein. The expression library described here, therefore, provides a powerful means to affinity select for B. japonicum genes encoding extracytoplasmic proteins. In total, 182 DNA sequences were analyzed, among which 132 different putative extracytoplasmic proteins could be identified. The function of most proteins could be predicted and support an extracytoplasmic localization. In addition, genes encoding novel extracytoplasmic proteins were found. In particular, a novel family of small proteins has been identified that is characterized by a conserved pattern of four cysteine residues. PMID- 12906118 TI - RDH5 gene mutations and electroretinogram in fundus albipunctatus with or without macular dystrophy: RDH5 mutations and ERG in fundus albipunctatus. AB - The aim of this study was to analyze the RDH5 gene in patients with fundus albipunctatus with and without macular dystrophy, and correlate the identified mutations with the electrophysiological results. Twenty-one patients from 19 unrelated Japanese families with fundus albipunctatus were examined. Ten unrelated patients had macular dystrophy. In 18 patients, either a homozygous or a compound heterozygous mutation in the RDH5 gene was identified. The bright flash, mixed rod-cone ERG had a negative configuration with reduced a-wave amplitudes after a short period of dark-adaptation (20 or 30 min). After a prolonged dark-adaptation period (2 or 3 h), the waveform attained normal amplitudes in patients without macular dystrophy but the a-waves were still subnormal in patients with macular dystrophy. The photopic ERG responses were significantly reduced in patients with macular dystrophy, indicating that they also had cone dystrophy. The photopic ERGs were reduced in only some of the patients without macular dystrophy. In patients without macular dystrophy, the scotopic b-wave amplitudes were nonrecordable or significantly reduced after a short dark-adaptation period but then improved to normal levels. However, they did not fully recover in some patients with macular dystrophy. Three patients with macular dystrophy in whom a RDH5 gene mutation could not be detected by our routine method had atypical ERG responses. We conclude that RDH5 gene mutations cause a progressive cone dystrophy or macular dystrophy as well as night blindness. The clinical phenotype including electrophysiological responses varied among patients with the RDH5 gene mutations. PMID- 12906120 TI - MfERG waveform characteristics in the RS1h mouse model featuring a 'negative' ERG. AB - Several retinal disorders lead to a relatively greater attenuation of the b-wave compared to the a-wave of the electroretinogram (ERG), a constellation called 'negative' ERG. To determine the waveform characteristics of multifocal ERGs (mfERGs) and their dependence on recording parameters in such a case, we studied the Rs1h(-/Y) mouse, the model for x-linked juvenile retinoschisis. mfERGs were recorded with a VERIS 4 system connected to a piggyback stimulator prototype that added the stimulus to the optical pathway of a HRA scanning-laser ophthalmoscope (SLO) by means of a wavelength-sensitive mirror. Real-time fundus visualization was achieved with the infrared laser of the SLO (835 nm). High-pass filter settings and the time interval used by the 'artefact removal' feature were varied to study their influence on the waveform. The mfERG in the Rs1h(-/Y) mouse had a 'negative' shape. However, the high-pass filter setting had to be lowered from the usual 10 Hz down to about 2 Hz in order to obtain that result, otherwise the negative shape was lost and mainly a positive peak remained. Similarly, a short time interval used by the 'artefact removal' feature also removed the negative shape. The Rs1h(-/Y) mouse was found to be a valuable model of diseases with a 'negative' waveform shape also in mfERG. Our results underline the importance of a lower high-pass filter cutoff frequency when recording mfERGs in such disorders. In addition, if the 'artefact removal' feature is used, it should be verified that it doesn't distort the waveform shape. PMID- 12906121 TI - Effects of telazol and nembutal on retinal responses. AB - PURPOSE: Compare the effects of Nembutal, a barbiturate, and Telazol, a dissociative anesthetic, on photoreceptor and post-receptoral retinal responses. METHODS: Dark adapted infant and adult albino rats were anesthetized with intraperitoneal injection of Nembutal or Telazol. ERG responses to full-field stimuli were recorded over a 6-8 log unit range from dim intensities at which the scotopic threshold response (STR) was observed to those that saturate the a-wave. The rod photoresponse, b-wave, oscillatory potentials (OPs), and STR parameters of rats in the Nembutal and Telazol groups were compared. RESULTS: For both infants and adults, the saturated amplitudes of the photoresponse and the b-wave were larger in Telazol than Nembutal rats, but sensitivity of the photoresponse did not differ significantly between Telazol and Nembutal rats. The STR was seen only in the Telazol responses of adults. There was little differential effect of the two agents on the OPs. CONCLUSIONS: Photoreceptor and postreceptoral responses recorded under Nembutal and Telazol anesthesia differ significantly. These results may inform selection of anesthetic for studies of animal models. PMID- 12906122 TI - A potential spontaneous rat model of X-linked congenital stationary night blindness. AB - PURPOSE: To describe a possible spontaneous rat model of X-linked congenital stationary night blindness (CSNB). METHODS: Experimental animals were generated by mating the affected animal to normal rats, and from interbreeding littermates. To define the inheritance pattern, full-field electroretinograms (ERGs) were recorded from all progeny. RESULTS: During the course of other experiments, an affected male rat was identified by a reduced amplitude ERG b-wave. When this rat was mated to normal Sprague-Dawley rats, all of the F1 progeny had normal ERG waveforms. When F1 offspring were interbred, 51% of the male offspring had b-wave reductions while all female offspring had normal ERG waveforms. When F1 females were backcrossed to the original affected male, b-wave reductions were noted in both male and female offspring; overall, 46.8% of the backcross progeny exhibited a b-wave reduction. In affected animals, the b-wave was selectively affected as the a-wave appeared to retain normal amplitude and kinetics at 1-4 months old. Cone ERGs were significantly reduced in amplitude and somewhat delayed. Similar ERG results were also obtained under the same stimulus conditions from human patients with complete CSNB (CSNB1). CONCLUSIONS: The inheritance pattern is consistent with an X-linked recessive trait. The electrophysiological results suggest that this mutant rat line may provide another model for CSNB1. PMID- 12906123 TI - Spontaneous occurrence of a potentially night blinding disorder in guinea pigs. AB - Several hereditary retinal disorders such as retinitis pigmentosa and congenital stationary night blindness compromise, sometimes exclusively, the activity of the rod pathway. Unfortunately, there are few animal models of these disorders that could help us better understand the pathophysiological processes involved. The purpose of this report is to present a pedigree of guinea pigs where, as a result of a consanguineous mating and subsequent selective breeding, we developed a new and naturally occurring animal model of a rod disorder. Analysis of the retinal function with the electroretinogram reveals that the threshold for rod-mediated electroretinograms (ERGs) is significantly increased by more than 2 log-units compared to that of normal guinea pigs. Furthermore, in response to a suprathreshold stimulus, also delivered under scotopic condition, which yield a mixed cone-rod response in normal guinea pigs, the ERG waveform in our mutant guinea pigs is almost identical (amplitude and timing of a- and b-waves) to that evoked in photopic condition. The above would thus suggest either a structural (abnormal development or absence) or a functional deficiency of the rod photoreceptors. We believe that our pedigree possibly represents a new animal model of a night blinding disorder, and that this condition is inherited as anautosomal recessive trait in the guinea pig population. PMID- 12906125 TI - Gene transfer in the RPE65 null mutation dog: relationship between construct volume, visual behavior and electroretinographic (ERG) results. AB - In vivo gene transfer in a large group of RPE65 null mutation dogs have been recently performed. The present study was aimed at determining, through visual behavioral and electroretinographic (ERG) testing, if there is a volume effect of the gene construct administered. Eleven Beagle-Briard dogs homozygous for the RPE65 null mutation and two unaffected control dogs were included. Affected animals were unilaterally treated with either a high (70-100 micro1; N = 6) or a low volume (30-60 microl; N = 5) of subretinally injected rAAV.RPE65 construct, at the age of 4 months to 2.5 years. Fellow eyes were treated with a subretinal injection of rAAV.GFP or sham operated and used as internal controls. Retinal function was measured pre- and 10-12 weeks post-surgically, using simultaneous bilateral full-field flash ERGs. A significant improvement in all ERG responses studied was identified for the high volume treated group compared to pre-surgical parameters. A significant improvement for the high intensity scotopic a-wave response for the low volume rAAV.RPE65 treated group was also found. Objective and subjective dim and day light visual maze testing, in eight of the affected treated animals, and the two control dogs, revealed better vision in daylight than in dim light for all animals. Vision in dogs treated with the high volume of gene construct was significantly better in day light than in dim light. No significant difference was noted between day and dim light testing for the control group or those animals treated with a low volume of the gene construct. Significantly better vision was noted in the control group when compared with the low volume group under dim light conditions, and the high volume group under day light conditions. No significant difference in functional vision could be identified between the high volume treated animals and control animals in day light conditions. These findings support the hypothesis that functional vision is improved by subretinal rAAV.RPE65 injection in a volume-dependent manner. PMID- 12906124 TI - Retinal defects in the zebrafish bleached mutant. AB - The recessive zebrafish mutant bleached has, apart from its defects in pigmentation, a heritable defect leading to larval blindness. Here, we analyze the retina of homozygous bleached larvae, employing morphological and electrophysiological methods. Electroretinography revealed a complete lack of electrical signals in response to light. Histological analysis of mutant retinae showed a severely affected outer retina with a hypopigmented pigment epithelium and a disorganized outer nuclear layer containing few or no intact photoreceptors. Using the TUNEL assay for cell death detection, we noticed a strong increase of apoptotic cells in all retinal cell layers, starting in young larvae even before retinal support of visual function. At later stages cell death is most pronounced at the marginal zone, where new cells are constantly added to the retina. At early stages increased apoptosis is mainly confined to the retina, while at later stages elevated cell death is al so apparent in extra-retinal tissues, particularly in the brain. Hence, the lack of visual responses in homozygous bleached larvae can be attributed to a severe defect of the outer retina, preceded by increased levels of apoptotic cell death in all retinal cell layers. PMID- 12906127 TI - Identification and chromosomal localization of mariner-like elements in the cabbage moth Mamestra brassicae (Lepidoptera). AB - A complete mariner-like element has been identified in the lepidopteran Mamestra brassicae. This element, called Mbmar, represents a new type of mariner transposon. It has a transposase similar to that of other insect mariner coding sequences but its inverted terminal repeats differ from typical mariner ones. This observation is unique since generally both mariner coding region and ITRs are evolutionarily conserved in insects. Mbmar is detectable by FISH only in the heterochromatic regions of both the sex chromosomes. PMID- 12906119 TI - Electrophysiological analysis of visual function in mutant mice. AB - The mouse has become a key animal model for ocular research. This situation reflects the fact that genes implicated in human retinal disorders or in mammalian retinal function may be readily manipulated in the mouse. Visual electrophysiology provides a means to examine retinal function in mutant mice, and stimulation and recording protocols have been developed that allow the activity of many classes of retinal neurons to be examined and which take into account unique features of the mouse retina. Here, we review the mouse visual electrophysiology literature, covering techniques used to record the mouse electroretinogram and visual evoked potential, and how these have been applied to characterize the functional implications of gene mutation or manipulation in the mouse retina. PMID- 12906126 TI - Molecular and cytogenetic organization of the 5S ribosomal DNA array in chicken (Gallus gallus). AB - The 5S ribosomal (r) RNA genes encode a small (approximately 120-bp) highly conserved component of the large ribosomal subunit. The objective of the present research was to study the molecular and cytogenetic organization of the chicken 5S rDNA. A predominant 2.2-kb gene (5Salpha) consisting of a coding and intergenic spacer (IGS) region was identified in ten research and commercial populations. A variant gene repeat of 0.6kb (5Sbeta) was observed in some of the populations. Genetic linkage analysis and cytogenetic localization by fluorescence in-situ hybridization assigned the 5S rDNA to chromosome 9. The 5S rDNA array was determined to be 80.2 +/- 7.0 kb upon electrophoretic sizing following EcoRV digestion. Sequence analysis of 5Salpha IGS regions revealed considerable conservation between chicken subspecies (98.4% identity) as well as homology with vertebrate Pol III promoter and regulatory sequence motifs. Minor intraindividual sequence variation within 1000 bp of IGS was observed in four cloned Red Jungle Fowl (Gallus gallus gallus) 5Salpha repeats (95.5% identity in this region). Sequence comparisons between IGS regions of 5Salpha and 5Sbeta genes indicated two short continuous (>20bp) and many short non-continuous homologous regions as well as other conserved features such as promoter and termination motifs. PMID- 12906128 TI - Human chromosome 16 conservation in primates. AB - A study was made of the organization of the chromosome orthologous to HSA16 in primates using a panel of 8 BAC probes spanning human chromosome 16. The probes were used in FISH experiments on great apes and on representatives of the Old World monkeys, New World monkeys, and lemurs. The domestic cat was used as an outgroup. The results indicate that 16p and 16q were separate chromosomes in a primate ancestor. They fused in a Catarrhini ancestor giving rise to the present day form found in HSA, great apes, and Old World monkeys. Several rearrangements were found in New World monkeys. PMID- 12906129 TI - Chromosomal studies in Callicebus donacophilus pallescens, with classic and molecular cytogenetic approaches: multicolour FISH using human and Saguinus oedipus painting probes. AB - This paper presents the karyotype of Callicebus donacophilus pallescens for the first time. The analysis included G-, C-, NOR-banding techniques and FISH with chromosome painting probes from Saguinus oedipus and Homo sapiens. The results were compared with the karyotypes of Callicebus moloch donacophilus and C. moloch previously published. These three karyotypes display the same diploid number (2n = 50) but diverge about the number of biarmed and acrocentric chromosomes. The acrocentrics 14 and 15 from C. m. donacophilus and C. moloch have undergone an in tandem fusion originating a large acrocentric (pair 10) in C. d. pallescens. The major submetacentric pair (pair 1) from C. d. donacophilus and C. moloch have undergone fission originating two acrocentric pairs in C. d. pallescens (pairs 15 and 22). Herein was evidence that, in spite of the high interspecific variation among Callicebus, most of the chromosomes remained conserved. PMID- 12906130 TI - Characterization of the atypical karyotype of the black-winged kite Elanus caeruleus (Falconiformes: Accipitridae) by means of classical and molecular cytogenetic techniques. AB - The karyotype of the black-winged kite (Elanus caeruleus), a small diurnal raptor living in Africa, Asia and southern Europe, was studied with classical (G-, C-, R banding, and Ag-NOR staining) and molecular cytogenetic methods, including primed in-situ labelling (PRINS) and fluorescence in-situ hybridization (FISH) with telomeric (TTAGGG) and centromeric DNA repeats. The study revealed that the genome size, measured by flow cytometry (3.1 pg), is in the normal avian range. However, the black-winged kite karyotype is particularly unusual among birds in having a moderate diploid number of 68 chromosomes, and containing only one pair of dot-shaped microchromosomes. Moreover, the macrochromosomes are medium-sized, with the Z and W gonosomes being clearly the largest in the set. C-banding shows that constitutive heterochromatin is located at the centromeric regions of all chromosomes, and that two pairs of small acrocentrics and the pair of microchromosomes are almost entirely heterochromatic and G-band negative. The distribution pattern of a centromeric repeated DNA sequence, as demonstrated by PRINS, follows that of C-heterochromatin. The localization of telomeric sequences by FISH and PRINS reveals many strong telomeric signals but no extratelomeric signal was observed. The atypical organization of the karyotype of the black winged kite is considered in the context of the modes of karyotypic evolution in birds. PMID- 12906132 TI - Pericentromeric euchromatin is conserved in minute human supernumerary chromosomes: a study using cross-species colour segmenting (RxFISH). AB - Investigation of marker chromosomes is one of the most challenging areas of clinical cytogenetics, especially in the prenatal scenario. A range of techniques including microdissection/reverse painting, SKY and M-FISH are available for the investigation of larger markers (>3 Mb). All these techniques rely on hybridization of unique, homologous sequences with simultaneous suppression of repeat sequences. In contrast, RxFISH is based on hybridization of cross-species syntenic sequences; repeat sequences do not hybridize due to species divergence. We have used RxFISH to analyse a group of the smallest, i.e. minute, supernumerary marker chromosomes. Our results suggest that even the smallest marker chromosomes often contain conserved pericentric euchromatin. More detailed characterization of pericentric genetic content is needed to assess the clinical significance of minute supernumerary markers. PMID- 12906131 TI - Partially functional Cenpa-GFP fusion protein causes increased chromosome missegregation and apoptosis during mouse embryogenesis. AB - CENP-A is an essential histone H3-like protein that localizes to the centromeric region of eukaryotic chromosomes. Heterozygous and homozygous Cenpa-GFP fusion protein mouse mutants, generated through targeted insertion of the green fluorescent protein (GFP) gene into the mouse Cenpa gene locus, show specific localized fluorescence at all the centromeres. Heterozygous mice are healthy and fertile. Cenpa-GFP homozygotes (Cenpag/g) undergo many cell divisions, giving rise to up to one million cells that show relatively accurate differentiation into distinct mouse embryonic tissues until day 10.5 when significant levels of chromosome missegregation, aneuploidy and apoptosis result in death. Cenpag/g embryos assemble functional kinetochores that bind to a host of centromere specific structural and mitotic spindle checkpoint proteins (Cenpc, BubR1, Mad2 and Zw10). Examination of the nucleosomal phasing of centromeric minor and pericentromeric major satellite sequences indicates that the formation of Cenpag/g homotypic nucleosomes is not accompanied by any overt alteration to the overall size of the monomeric nucleosomal structure or the spacing of these structures. This study provides the first example of an essential centromeric protein gene variant in which subtle perturbation at the centromeric nucleosomal/chromatin level manifests in a significantly delayed lethality when compared with Cenpa null mice. PMID- 12906135 TI - Isolation and identification of cDNA fragments and full-length cDNA differentially expressed in human glioblastoma cell line BT-325 versus all-trans retinoic acid induction . AB - OBJECTIVE: To investigate the differentiation process of the human glioblastoma cells. METHODS: Differential display reverse transcribed-PCR (DDRT-PCR) was used to isolate the genes differentially expressed in control and all-trans retinoic acid treated human glioblastoma cell line BT-325. Routine method of cDNA library screening was performed to clone full-length cDNA. RESULTS: Thirty-six RT-PCR reactions were performed and 64 differentially expressed fragments were recovered, amplified and cloned. Of them, 46 ESTs were sequenced and delivered into the GenBank. The homology comparison using BLAST algorithm revealed that 22ESTs are highly homologous with the known genes and many of them play important roles in the cell differentiation progress. A dot-blot hybridization was conducted to certify the differentiation expression. The result showed that 27 EST clones are expressed at different level in control and all-trans retinoic acid treated BT-325 cells. A full-length cDNA was cloned using the EST-HGBB098. CONCLUSION: DDRT-PCR was a simple and effective method to serially analyze the differentially expressed genes. PMID- 12906133 TI - Identification and characterization of U1 small nuclear RNA genes from two crustacean isopod species. AB - Four different units containing three variants of the U1 snRNA gene have been identified in the genome of Asellus aquaticus and only one unit has been identified in the genome of Proasellus coxalis. All four identified U1 snRNA genes can be folded according to the proper secondary structure and possess the functionally useful conserved sequences. Moreover, in the 3 flanking regions, all genes present both the 3 box, a conserved sequence required for 3 processing of mature snRNA, and a polyadenylation signal which is unusual for these genes. The PCR products were used as probes in fluorescent in-situ hybridization (FISH) experiments to locate them on chromosomes of A. aquaticus and P. coxalis. PMID- 12906136 TI - The clinical evaluation of the effects of a new collagenase ointment (Iruxol mono) on debridement and wound healing in the burn wounds. PMID- 12906134 TI - Population variation in the A chromosome distribution of satellite DNA and ribosomal DNA in the grasshopper Eyprepocnemis plorans. AB - The double FISH analysis of two repetitive DNAs (a satellite DNA and ribosomal DNA) in 12 natural populations of the grasshopper Eyprepocnemis plorans collected at the south (Granada and Malaga provinces) and south-east (Albacete and Murcia provinces) of the Iberian Peninsula has shown their wide-spread presence throughout the whole genome as well as extensive variation among populations. Both DNAs are found in most A chromosomes. Regularly, both DNAs occurred in the S11 and X chromosomes, rDNA in the S10 and satDNA in the L2 and M3. No correlation was found between the number of satDNA and rDNA clusters in the A genomes of the 12 populations analysed, and both figures were independent of the presence of B chromosomes. The genomic distribution of both DNAs showed no association with the geographical localization of the populations analysed. Finally, we provide evidence that the supernumerary chromosome segment proximally located on the S11 chromosome is, in most cases, the result of satDNA amplification but, in some cases, it might also derive from amplification of both satDNA and rDNA. PMID- 12906137 TI - Experimental study of homium: YAG laser transmyocardial revascularization in acute ischemic settings with myocardial contrast echocardiography. AB - OBJECTIVE: To study the mechanism and effects of blood perfusion to the acute ischemic region of myocardium through Ho-YAG laser channels with myocardial contrast echocardiography. METHODS: To produce the model of acute myocardial ischemia, we partially ligated the left anterior decending (LAD) coronary artery of canine hearts between 1st. and 2nd. diagonal branches and then performed transmyocardial revascularization in this region with Ho - YAG laser. Myocardial contrast echocardiography was made with a new generation of ultrasound contrast agent and second harmonic imaging of this region before, after ischemia and after laser revascularization. Pictures were taken with "R" wave trigger skill. RESULTS: Acoustic density determing in the ischemia region (anterior wall) with MCE (myocardial contrast echocardiography) was obviously decreased (5.40 +/- 1.81) after the LAD was ligated,as compared with before (11.69 +/- 1.61, P < 0.01). It was increased remarkably after transmyocardial laser revascularization (TMLR) (11.2 +/- 2.01, P < 0.01) as compared with that when ischemia and approximated to that before ischemia (P > 0.05). There were no differences in acoustic density in the lateral wall (as control) among these comprehensive three periods (P > 0.05). Contrast in the laser region developed one cardiac cycle ahead of that in the non-ischemic normal region. CONCLUSION: Acute ischemic myocardium can be perfused by oxygenated blood from the left ventricle through Ho - YAG laser channels. Evidence of blood perfusion through laser channels during systolic phase was detected, and myocardial contrast ultrasonography using intravenous perfluorocarbon-exposed sonicated dextrose albumin may be regarded as a reliable method in the study of transmyocardial revascularization. PMID- 12906138 TI - Serum levels of vascular endothelial growth factor in patients with angina pectoris and acute myocardial infarction. AB - OBJECTIVE: To determine whether serum vascular endothelial growth factor (VEGF) concentrations are altered in several kinds of coronary heart disease patients. MATERIALS AND METHODS: Using a VEGF enzyme-linked immunosorbent assay (ELISA), serum VEGF concentrations were determined in antecubital venous blood of 16 patients with stable angina pectoris (SAP), 16 with unstable angina pectoris (UAP) and 16 with acute myocardial infarction (AMI) before and after thrombolytic therapy, and of 16 age- and sex-matched healthy volunteers who used as controls. RESULTS: The concentrations of serum VEGF in patients with SAP (98.60 +/- 26.99 pg/ml) and UAP (103.61 +/- 24.89 pg/ml) tended to be higher than those in control subjects (80.44 +/- 24.57 pg/ml), but the differences did not reach statistical significance (P > 0.05 for each). Before thrombolytic therapy, the concentrations of serum VEGF in patients with AMI (285.92 +/- 125.15 pg/ml) were significantly higher than those in patients with SAP, UAP or control subjects (P < 0.01, respectively), and correlated with synchronous serum creatine kinase (CK) and its MB isoenzyme (CK-MB) contents (r = 0.866, P < 0.001 and r = 0.948, P < 0.001; respectively). Three hours after thrombolysis, the concentrations of VEGF had fallen to 111.57 +/- 31.29 pg/ml (P < 0.01 vs x before thrombolytic therapy and P < 0.05 vs x control subjects). CONCLUSION: The present study shows that serum concentrations of VEGF in patients with AMI are markedly elevated and that increased serum VEGF levels may be one of the most sensitive indexes in diagnosing AMI and judging reperfusion. PMID- 12906139 TI - The effect of basic fibroblast growth factor slow-release microcapsules on angiogenesis in infarcted rabbit myocardium. AB - OBJECTIVES: To observe the effect of basic fibroblast growth factor (bFGF) slow release microcapsules on angiogenesis in infarcted myocardial regions. METHODS: Myocardial infarction was induced in 24 New Zealand rabbits by ligating the root of left anterior descending coronary artery. Group I (n = 8) served as control, group II (n = 8) as a blank microcapsule group, group III (n = 8, each microcapsule contains 1 microg bFGF) as microcapsule group. In group II and III, 5 blank microcapsules or bFGF slow-release microcapsules were implanted into myocardium underneath the epicardium between the left anterior descending coronary artery and left circumflex artery. Infarct size was evaluated by infarcted weight/left ventricle weight ratio and angiogenesis was evaluated by immunohistochemical examinations 5 weeks later. RESULTS: As compared with group I and II, rabbits treated with bFGF slow-release microcapsules showed higher microvessel counts (group I 37.75 +/- 4.50, group II 38.37 +/- 4.98, vs x group III 135.50 +/- 4.81, P < 0.001) and less infarcted weight/left ventricle weight (group I 16.8% +/- 0.4%, group II 16.7% +/- 0.5%, vs x group III 7.0% +/- 0.2%, P < 0.001). CONCLUSIONS: Subepicardial administration of bFGF slow-release microcapsule in the infarcted rabbit model results in effective angiogenesis and reduction in infarct size. PMID- 12906140 TI - The effect of left ventricular diastolic dysfunction on the pathogenesis of angina decubitus. AB - OBJECTIVE: To investigate the effect of left ventricular diastolic dysfunction on the pathogenesis of angina decubitus (AD). METHODS: The study population consisted of three groups: 20 individuals without cardiovascular studied as group I. Group II included 20 patents with coronary artery disease and without AD. Thirty-one patients with AD and ejection fraction (EF) > 50% were studied as group III. Group II and III were matched for age, EF and extent of coronary artery disease. RESULTS: Left ventriculography (LVG) showed that left ventricular (LV) first 1/3 filling fraction (1/3FF) was significantly lower in group III than in group II and I (both P < 0.001), but LV late 1/3 FF was much higher in group III than in group II and I (P < 0.05, P < 0.01). Left ventricular end-diastolic pressure (LVEDP) was markedly increased before and after LVG in group II and III as compared with group I (both P < 0.05, both P < 0.001). The difference of LVEDP caused by left atrial contraction (left atrial contraction pressure difference, LACPD) before and after LVG was much higher in group III than in group I ( P < 0.01, P < 0.001). However, there we significant differences in LVEDP and in LACPD between before and after LVG only in group III (both P < 0.01). CONCLUSION: The patients with AD have LV diastolic dysfunction, which may be closely related to the pathogenesis of angina decubitus. PMID- 12906141 TI - Nitric oxide inhibits a rise of ATP-introduced cytosolic free Ca2+ concentration and release from intracellular stored Ca2+. AB - OBJECTIVE: The effects of ATP-introduced a rise in cytosolic free Ca2+ concentration and inhibition of nitric oxide were investigated. METHOD: Measurement of free Ca2+ ([Ca2+]i) of cultured rat tail arterial smooth muscle cells using Fura-2/AM dual excitation wavelength spectrofluorometer. RESULTS: There are two components of [Ca2+]i can be evoked by ATP. One part is Ca2+ entry from Ca2+ channel and formed a plateau. The another part is a peak that released from Ca2+ store. Both of them can be inhibited by NO. CONCLUSION: The ATP induced [Ca2+]i rise that release Ca2+ from both Insp3 and ryanochine receptors and Ca2+ entry through calcium channels. The inhibition of NO on ATP induced [Ca2+]i rise that was mediated by cGMP. PMID- 12906143 TI - Calorie restriction can increase thymocyte apoptosis through Bcl-2 and Fas pathway. PMID- 12906142 TI - Complications in laparoscopic gynecologic surgery. AB - OBJECTIVE: To investigate retrospectively the complications and associated factors of gynecolgical laparoscopies. METHODS: 1 769 laparoscopic surgeries were carried out from January 1994 to October 1999 at our department. The procedures included 1421 surgeries of ovary and tube, 52 myomectomies and 296 cases of laproscopic-assisted vaginal hysterectomy (LAVH). A total of 312 patients had a history of prior laparotomy (17.6%). RESULTS: Complications occured in 34 cases, the overall complication rate was 1.92%. Unintended laparotomies occured in 6 cases (0.34%). 12 complications were associated with insertion of Veresse needle or trocar and creation of pneumoperitoneum, including 5 severe emphysema and 7 vascular injuries, this figure represents 35.3% of all complications of this series. Five intraoperative complications (14.7%) occured during the laparoscopic surgery (3 severe bleedings, one bladder injury and one skin burn of leg caused by damaged electrode plate), laparotomy was required in four of these cases. Seventeen complications occured during postoperative stage: 2 intraperitoneal hemorrhages needing laparotomy, 2 bowel injuries, 4 nerve paresis and 9 febrile morbidities. CONCLUSIONS: Operative gynecologic laparoscopy is associated with acceptable morbidity rate, but can not be overlooked. Complication rate seems to be higher in advanced procedures such as LAVH. PMID- 12906144 TI - The treatment of lumbosacral instability by the Diapason system. AB - OBJECTIVE: To introduce a new internal fixation system of spine and its characteristics. METHODS: To review 16 patients with lumbosacral instability who were their clinical outcomes and radiographic evaluation. RESULTS: Fifteen patients gained complete recovery from their preoperative symptoms. One patient who had experienced two operations before and with problems of urinary and fecal incontinence and walking difficulty still had lower limb pain, muscle weakness and urinary incontinence after operation. There is no evidence of spine glide on X-ray, implant failure, neural complication or infection during follow up. CONCLUSION: Diapason system can achieve good early postoperative results with few complications and ease to use for lumbosacral instability. PMID- 12906145 TI - An epidemiology and molecular genetic study on breast cancer susceptibility. AB - OBJECTIVES: To investigate the genetic susceptibility for breast cancer of Chinese, a hospital-based case-control study, pedigree survey and molecular genetic study were conducted. METHODS: Logistic regression model and stratification methods were used in the risk factors analysis. Li-Mantel-Gart and Falconer methods were used to analyse the segregation ratio and heritability. Polymerase chain reaction (PCR) and polyacrylamide gel electrophoresis were used to detect AI, G-binding technique was used to detect the chromosome aberration of peripheral blood lymphocyte. RESULTS: Family history of breast cancer is related to enhanced breast cancer risk significantly, OR is 3.905 (95% CI = 1.079 approximately 14.13), and it widely interacts with other risk factors. Accumulative incidence of breast cancer in first degree relatives is 9.99%, which is larger than that in second, third degree and non-blood relatives. Segregation ratio is 0.021, heritability among first degree relatives is 35.6 +/- 5.8%. Frequencies of LOH at BRCA1 and BRCA2 loci in sporadic breast cancer are 6.12% and 5.77% respectively. In the sibs, both of them show LOH at D13S173 locus, and high frequencies of chromosome aberrations were observed. CONCLUSIONS: Genetic susceptibility contributes to breast cancer occurrence of Chinese, and its racial variation may be one of the important reasons for the large difference of incidence between western and eastern countries. PMID- 12906147 TI - Adenosquamous lung carcinoma: clinical characteristics, surgical treament and prognosis. AB - OBJECTIVE: The effectiveness of surgical resection of adenosquamous carcinoma of the lung remains poorly defined because of the histology's relatively low frequency, the failure in most published series to separate adenosquamous carcinoma from the other variants of non-small cell lung carcinoma. To define the effectiveness of treatment of adenosquamous carcinoma, we have retrospectively reviewed our hospital experience over a 12-year period. METHODS: Retrospectively reviewed 22 cases of adenosquamous carcinoma who were surgically treated, except one patient, in the PUMCH from Jan. 1985 to Aug. 1997. This series constitutes the 1.9% of a total of 1 245 patients with all types of surgical treatment for the primary lung cancer during the same time. RESULTS: The adenosquanous carcinoma was mostly presented in the old patients with a mean age of 60 years and mostly located in the peripheral of lung (n = 20). The overall 5-year survival was 23%. Those with stage I tumors survival was only 18% (n = 13), stage II 5%. The survival in stage III tumos was not longer than 25 months and in stage IV survival was not longer than 12 months. CONCLUSION: Our results suggest that adenosquamous carcinoma of lung was a virulent tumor, which exhibited highly aggressive biological behavior with early lymph nodes metastasis (46%) and its prognosis was worse than that of both squamous cell carcinoma and adenocarcinoma. PMID- 12906146 TI - The study of mechanism of vascular endothelial growth factor on the prevention of restenosis after angioplasty. PMID- 12906149 TI - Treatment of sodium disorders after surgery of craniopharyngioma. AB - OBJECTIVE: To study the treatment of the patients with blood sodium disorder after craniopharyngioma surgery. METHOD: The blood sodium in 44 patients with craniopharyngiomas was daily examined from operative to post-operative days. Hypernatremia is defined as [Na] > 145 mmol/l, and hyponatremia as [Na] < 135 mml/L. RESULTS: Of the 44 patients, 36 developed sodium disorder. Among them, simple hyponatremia was 16 patients, simple hypernatremia was 9 patients, and alternative sodium disorder was 11 patients. CONCLUSION: The blood sodium disorder in patients with craniopharyngiomas after surgery appears to present complicated changes. Three types of blood sodium disorder could be determined as simple hyponatremia, simple hypernatremia, and alternative sodium disorder. Treatment of the sodium disorder in patients with after surgery could be carried in accordance with the above three types. PMID- 12906148 TI - Difference of rejection in single versus combined pancreas and kidney transplantation in rats. AB - OBJECTIVE: To investigate the difference of rejection in single versus combined pancreas and kidney transplantation in rats. METHODS: Allograft models including simultaneous pancreas and kidney (SPK) transplant and pancreas or kidney transplant alone were established in SD-Wistar rats, rejections of pancreas and kidney in different models were compared morphologically and functionally. RESULTS: Mean survival time (MST) of pancreas was significantly prolonged in SPK than in pancreas transplant alone (PTA) (11.5 days vs. 9.2 days, P < 0.05). Incidence of interstitial pancreatic rejection at gade II and grade III was much obvious in PTA than in SPK (42.9% vs. 12.5% at grade II and 28.6% vs 6.3% at grade II , P < 0.05). No significant difference was found in MST between SPK and kidney transplant alone( KTA). Administration of cyclosporine A prolonged the MST of pancreas and kidney, without altering the tendency stated above. CONCLUSIONS: In SPK, the function of pancreas is protected by kidney hence the severity of rejection is reduced, whereas the function of kidney is not protected by pancreas. It suggests that different organs differ in immunoallergization and immunoregulation, and immune response tend to attack organs with greater immunoactivity, those organs with minor one could be protected. Cyclosporine A is effective on prolonging the MST of pancreas and kidney. PMID- 12906150 TI - Diagnosis and treatment of insidious recurrent small intestinal hemorrhage. AB - OBJECTIVE: To improve the localized diagnosis of insidious recurrent small intestinal hemorrhage. METHODS: This retrospective analysis include 64 cases of such diseases, which were admitted from 1988 to 1998 to our hospital. RESULT: Ultrasonography, CT, small bowel pneumobariumgraphy, diluted barium enema, isotopic examination, DSA and intraoperative small-bowel endoscopy were used for diagnosis of hemorrhagic site, and 37 cases got a definite location before operation, while 10 cases were confined the diagnosis during the operation. Forty seven cases were treated surgically, while the other 17 cases had non-surgical treatment. Of the 47 cases, 39 cases underwent partial enterectomy, 5 cases had suture and ligature of vascular deformity, 2 cases had Whipple's operation, and one patient had ectomy of the end of ileum and right colon. CONCLUSION: DSA, Isotopic examination and intraoperative enteroscopy are of considerable importance for the location judgement of recurrent small intestinal hemorrhage. PMID- 12906151 TI - Long-term dietary restriction can decrease the alveolar macrophages to produce reactive oxygen intermediates and reactive nitrogen intermediates. PMID- 12906152 TI - The treatment of chylothorax. AB - OBJECTIVES: To understand and grasp the diagnosis and treatment of chylothorax caused by variou reasons. METHOD: The treatment results of 31 cases of chylothorax in PUMC hospital from 1963 approximately 1997 were retrospectively analyzed. RESULTS: Among 31 cases, 18 underwent surgery, 14 of 18 were cured, 2 died. In the 13 treated conservatively, 2 were cured, 3 died. Eleven cases were congenital, iatrogenic and traumatic chylothorax, 8 of them received surgical treatment and 6 of 8 were cured. The spontaneous chylothorax of unknown cause were 10 cases, 7 were treated by surgery and 6 were cured. CONCLUSION: Surgical intervention should be aggressively recommended for the traumatic, congenital, and iatrogenic chylothorx. The definite reason must be found out for the spontaneous chylothorax, corresponding management will be given according to the reason. Surgical ligation of the thoracic duct will contribute good result for the chylothorax of unknown cause, but combination of multiple treatment measures will be necessary for a successful management. PMID- 12906153 TI - On the sequence and function of kil gene of P1 phage. PMID- 12906154 TI - Influence of anti-TNF alpha monocolonal antibody on intestinal barrier in rats with acute pancreatitis. PMID- 12906155 TI - Cloning and analysis of the genomic DNA sequence of augmenter of liver regeneration from rat. AB - OBJECTIVE: To search for genomic DNA sequence of the augmenter of liver regeneration (ALR) of rat. METHODS: Polymerase chain reaction (PCR) with specific primers was used to amplify the sequence from the rat genome. RESULTS: A piece of genomic DNA sequence and a piece of pseudogene of rat ALR were identified. The lengths of the gene and pseudogene are 1508 bp and 442 bp, respectively. The ALR gene of rat includes 3 exons and 2 introns. The 442 bp DNA sequence may represent a pseudogene or a ALR-related peptide. Predicted amino acid sequence analysis showed that there were 14 different amino acid residues between the gene and pseudogene. ALR-related peptide is 84 amino acid residues in length and relates closely to ALR protein. CONCLUSION: There might be a multigene family of ALR in rat. PMID- 12906156 TI - The coexpression of the preS1 (1-42) and the core (1-144) antigen of HBV in E. coli. AB - OBJECTIVE: To study the therapeutic T cell vaccine for the treatment of chronic hepatitis B by improving the cellular immunization of HBsAg vaccine with the coexpression of the preS1 (1-42) and the Core (1-144) antigen of HBV in E. coli. METHODS: The genes of HBcAg (1-144) and preS1 (1-42) were amplified and fused by PCR. This fused gene was inserted in the prokaryotic expression vector pET-11d and expressed in E. coli. RESULTS: It was showed by SDS-PAGE that the protein molecular weight of the coexpression product was about 20 kD, 20% of all bacteria protein. The monoclonal antibodies against core and preS1 antibody could react with this fused protein by Western-blot technique respectively. The fused gene was verified by sequencing. Under the immune electron microscopy, this fused protein is typical particle of HBcAg but in an aggregated form. CONCLUSION: The results might aid for studying T cell immunotherapeutic vaccine for chronic hepatitis B. PMID- 12906158 TI - A survey of elective surgical patients' attitudes toward anesthesia in PUMC hospital. AB - OBJECTIVE: To assess patients' knowledge, attitudes, and concerns regarding anesthetic management. METHOD: A survey of 55 items was developed and administered preoperatively to 500 patients including 190 men and 310 women in our hospital. Patients were interviewed on their knowledge of the role of anesthesiologists, their preferences regarding anesthetic management, and also their concern about potential anesthetic complications. RESULTS: Patients' perceptions of anesthesiologists' training and role have reached a certain level. Most significant preoperative concerns regarding the anesthesiologists focused on experience, qualifications, and presence or absence during the anesthesia. Patients' concerns also included the possibility of not being waken up following anesthesia, experiencing postoperative pain, and becoming paralyzed. The variations of concerns depended partially on patients' sex, type of anesthesia, and proposed surgical procedure, partially on their education and living environments. CONCLUSION: It is suggested that anesthesiologist address significant patient concerns during the preoperative visit to enhance their effectiveness in patient care. Efforts to educate the public on the anesthesiologists' role in preoperative care would improve patients' confidence. PMID- 12906157 TI - Study on the relationship between mitochondrial gene mutation and latent autoimmune diabetes mellitus in adults. AB - OBJECTIVE: To identify the A3243G mutation of mitochondrial (mt) DNA in patients with latent autoimmune diabetes mellitus in adults (LADA) of Han nationality in the northeast area of China. METHODS: Seventy-nine diabetics of Han nationality, whose families have resided in the northeast area of China for more than 3 generations, were divided into 3 groups: Group 1 (22 cases of type 2 diabetes with maternal inheritance history), Group 2 (34 cases of LADA), Group 3 (23 cases of type 1 diabetes in adolescents). The A3243G of mt DNA was detected in these 79 subjects with the method of PCR-RFLP. RESULTS: None of the 79 diabetics studied was positively identified for the A3243G mutation of mt DNA. CONCLUSION: The A3243G mutation of mt DNA might not be related to the onset of LADA in diabetic population of Han nationality in northeast area of China and there might not be close relationship between A3243G mutation of mt DNA and autoimmunity. PMID- 12906159 TI - The construction and expression of recombinant shuttle plasmid with OmpL1 gene from leptospira interrogans serovar Lai strain 017 in Bacille Calmette-Guerin. AB - OBJECTIVE: To construct recombinant BCG against leptospirosis. METHODS: We amplified the entire open reading frame of the OmpL1 gene from the genome of the leptospire serovar Lai strain 017. Two recombinant plasmids pBQ1 and pBQ2 were constructed by oriented ligation based on the E. coli-BCG shuttle plasmids pMV261 and pMV361 respectively. The recombinant plasmids were transformed into BCG by electroporation. The rBCGs bearing pBQ1 and pBQ2 were induced by high temperature of 45 degrees C. RESULTS: The expressed product, a 35kD protein was detected by SDS-PAGE. The result indicates that pBQ1 and pBQ2 can express OmpL1 in rBCG. CONCLUSION: The technical methods in this study may help detect the immunogenicity and immunoprotection of OmpL1 and develop more safe, highly effective rBCG bearing leptospiral antigen with long-lasting protection. PMID- 12906160 TI - Experience of minimally invasive treatment in 520 patients with intracranial aneurysms. AB - OBJECTIVE: To summarize the experience of minimally invasive treatment in 520 patients with intracranial aneurysms on a retrospective study. METHODS: The measures used in the treatment of 520 patients were reviewed in terms of timing of surgery, induced-hypotensive anesthesia, brain protection combined with temporal occlusion of the feeding artery, external drainage of CSF, dynamic monitoring of intracranial pressure, blood flow velocity, serum osmolality and CT scanning, anti-vasospasm therapy as well as selected interventional endovascular embolization of aneurysms. RESULTS: Of the 520 patients, 485 were treated with either direct clipping or endovascular embolization and 35 patients were treated non-surgically. In 449 patients undergoing direct clipping and 36 undergoing endovascular embolization, intraoperative rupture of aneurysm occurred in 27 (6.0%) and 0%, respectively. Death occurred in 13 (2.6%), hemiplegia in 8 (1.6%), and vegetative state in 2 (0.4%). The operative mortality of direct clipping was 3.8% in 210 patients before 1990 and 1.8% in 275 patients after 1990 (36 patients undergoing endovascular embolization, the operative mortality was 0%). CONCLUSION: The outcome of patients with intacranial aneurysms can be markedly improved and the operative mortality can be lowered by minimally invasive treatment. PMID- 12906161 TI - Comparative study of fibronectin gene expression in tissues from hypertrophic scars and diabetic foot ulcers. AB - OBJECTIVE: To explore the expression characteristic of fibronectin gene in hypertrophic scars and diabetic ulcer tissues. METHODS: The biopsies from normal skins, hypertrophic scars and diabetic foot ulcers were taken. The technique of quantitative polymerase chain reaction (PCR) was used to evaluate the gene expression of fibronectin in the above biopsies. RESULTS: Fibronectin gene expression was enhanced in hypertophic scars and decreased in diabetic foot ulcers compared with that in normal skins. Quantitative comparison showed about 2 fold increase of fibronectin mRNA level in hypertrophic scars and about 3-fold decrease of fibronectin mRNA level in diabetic ulcers as compared with that in normal skins. CONCLUSIONS: Fibronectin gene expression is influenced by the tissue environment. Different expression and synthesis of fibronectin may cause different outcomes in wound healing. PMID- 12906162 TI - Intravascular ultrasound evaluating coronary stents for patients with coronary artery disease: compared old with new multilink stents. AB - OBJECTIVE: It was suggested that coronary stent design and coating may affect stent performance and hence induce varying degrees of thrombogenesis and neointimal hyperplasia. The purpose of this study is to compare the 6-month follow-up results between old and new Multilink stents with the method of intravascular ultrasound (IVUS) imaging. METHODS: We have performed old (n = 40) and new (n = 35) Multilink stent implantations on 75 patients with coronary artery disease. Coronary angiography was performed before, immediately after, and 6 months after the in-stent procedure respectively. Six-month follow-up IVUS imaging was performed and analyzed off-line. RESULTS: Minimal lumen cross sectional area (CSA) of new Multilink stents was significantly larger than that of old Multilink stents (P = 0.0053). Mean stent lumen area of new Multilink stents was significantly larger than that of old Multilink stents (P = 0.040). Similarly, minimal lumen diameter (MLD) of new Multilink stents was larger than that of old Multilink stents (P = 0.011). Old Multilink stents had a higher percentage of plaque area than new Multilink stents. CONCLUSION: The new Multilink stent is obviously superior to old Multilink stents, in particular, in the stent MLD and lumen CSA--major determinants of the restenosis. PMID- 12906163 TI - Methylation of p16 and p15 genes in multiple myeloma. AB - OBJECTIVE: To investigate the frequency of p16 and p15 gene methylation in multiple myeloma (MM), and its relationship with bone marrow cell apoptosis and clinical outcome. METHODS: Twenty-two patients with MM were studied to detect p16 and p15 gene methylation. Methylation-specific polymerase chain reaction (MSP) was used to detect gene methylation, and terminal transferase-mediated dUTP nick end-labeling (TUNEL) was used to detect cell apoptosis. RESULTS: p16 and/or p15 gene methylatoin was detected in 10 of 22 patients (45.4%). There were 3 patients with p16 gene methylation, 9 patients with p15 gene methylation, and 2 patients with both genes methylation. The incidence of methylation of p15 gene was higher than that of p16 gene (P < 0.05). The patients with p16 and/or p15 gene methylation had a delayed cell apoptosis, poor response to chemotherapy, and a short over-all survival (OS). CONCLUSION: The methylation of p16 and/or p15 gene plays a key role in MM apoptosis pathogenesis. The patients with both p16 and p15 gene methylation had a poor prognosis. PMID- 12906165 TI - The increase in plasminogen activator inhibitor type-1 expression by stimulation of activators for peroxisome proliferator-activated receptors in human endothelial cells. AB - OBJECTIVE: [corrected] To investigate the effect of peroxisome proliferator activated receptors (PPARs) activators on plasminogen activator inhibitor type-1 (PAI-1) expression in human umbilical vein endothelial cells and the possible mechanism. METHODS: Human umbilical vein endothelial cells (HUVECs) were obtained from normal fetus, and cultured conventionally. Then the HUVECs were exposed to test agents (linolenic acid, linoleic acid, oleic acid, stearic acid and prostaglandin J2 respectively) in varying concentrations with fresh media. RT-PCR and ELISA were applied to determine the expression of PPARs and PAI-1 in HUVECs. RESULTS: PPAR alpha, PPAR beta and PPAR gamma mRNA were detected by using RT-PCR in HUVECs. Treatment of HUVECs with PPARalpha and PPAR gamma activators- linolenic acid, linoleic acid, oleic acid and prostaglandin J2 respectively, but not with stearic acid could augment PAI-1 mRNA expression and protein secretion in a concentration-dependent manner. However, the mRNA expressions of 3 subclasses of PPAR with their activators in HUVECs were not changed compared with controls. CONCLUSION: HUVECs express PPARs. PPARs activators may increase PAI-1 expression in ECs, but the underlying mechanism remains unclear. Although PPARs expression was not enhanced after stimulated by their activators in ECs, the role of functionally active PPARs in regulating PAI-1 expression in ECs needs to be further investigated by using transient gene transfection assay. PMID- 12906164 TI - Apoptotic and proliferative activity in ovarian benign, borderline and malignant tumors. AB - OBJECTIVE: To determine the apoptotic and proliferative activities in various ovarian epithelial tumors. METHODS: Formalin-fixed, paraffin-embedded tissues of 86 ovarian epithelial tumors, including 52 adenocarcinomas, 23 borderline tumors and 11 cystadenoma, were retrieved. Apoptotic (AI) and proliferative (PI) index were estimated using the monoclonal antibodies: M30, Ki-67 and Ki-S1 in these tumors. Quantitative assessment of AI and PI was estimated by calculating the percentage of positive cells among no less than 1000 tumor cells. RESULTS: Statistically significant difference in AI was found between benign and borderline tumors or carcinomas (P = 0.028, 0.001, respectively). Significant differences in PI, as assessed by both Ki-67 and topo IIalpha, were demonstrated between carcinomas and benign or borderline tumors (both P < 0.001). Benign tumors had both low PI and AI; borderline tumors had lower PI but higher AI, while adenocarcinomas had both high proliferative and high apoptotic rates. Among borderline tumors, serious tumors had significantly lower AI and higher PI than mucinous ones. CONCLUSION: The results suggest that apoptotic and proliferative activities play important roles in the pathogenesis and development of ovarian borderline and malignant tumors. The high apoptotic rate in borderline tumor may explain its relatively indolent behavior while the high proliferative rate in carcinomas tends to explain its aggressive behavior. PMID- 12906166 TI - Peripartum cardiomyopathy--report of 16 cases. AB - OBJECTIVE: To analyze the clinical characteristics of peripartum cardiomyopathy and to evaluate the different factors that influence the prognosis of the peripartum cardiomyopathy. METHOD: A retrospective review was undertaken on records of women who were diagnosed with peripartum cardiomyopathy at Peking Union Medical College Hospital between Jan. 1983 and May 1999. RESULTS: During the research period, only 16 pregnant women were documented as peripartum cardiomyopathy. Some of the women undertook ultrasonic cardiographic (UCG) examination that showed decreased systolic function. Seven women were complicated with pregnancy induced hypertension. Three died of disseminated intravascular coagulation, embolism and cardiogenic shock respectively. CONCLUSION: Early diagnosis of the peripartum cardiomyopathy is extremely important. The UCG can provide helpful information on disease progression or regression. PMID- 12906167 TI - Serum soluble E-cadherin level in patients with endometriosis. AB - OBJECTIVE: To investigate the serum sE-cadherin level in patients with endometriosis and the alterations of that level in healthy control during the menstrual cycle. METHODS: Thirty-two patients with endometriosis and 30 healthy women were tested for serum sE-cadherin levels by enzyme-linked immunosorbent assay. RESULTS: The serum sE-cadherin levels in healthy control did not vary throughout the menstrual cycle, which were lower than those in patients with endometriosis. CONCLUSIONS: E-cadherin might be involved in endometrial shedding during menstruation in endometriosis patients. The serum sE-cadherin assay might be helpful as a serum marker for the diagnosis and management of endometriosis. PMID- 12906168 TI - A clinicopathological analysis of 22 cases of multiple malignant tumors. AB - To get a better understanding of the location, pathophysiology, etiology and prognosis of multiple malignant tumors (MPMT), we evaluated the medical records of 22 patients with MPMT. Our results suggested that radiotherapy and chemotherapy might play an important role in the pathogenesis of MPMT and follow up is important in detecting a secondary primary malignant tumor (PMT) at an early stage. Surgical removal of tumors is the first-choice therapy for MPMT. PMID- 12906169 TI - The effects of nerve growth factor on skin healing and blood recovery in irradiated mice. PMID- 12906171 TI - Next-generation x-ray CT units will provide <500 msec images with 3D resolution comparable to today's projection radiography. Against the proposition. PMID- 12906170 TI - Next-generation x-ray CT units will provide <500 msec images with 3D resolution comparable to today's projection radiography. For the proposition. PMID- 12906172 TI - Which accelerator photon beams are "clinic-like" for reference dosimetry purposes? AB - Previous work has demonstrated that, for photon beam dosimetry, TPR10(20) is not an ideal beam quality specifier for all bremsstrahlung beams, especially for lightly filtered beams in some standards laboratories. This paper addresses the following questions: Is TPR10(20) an adequate beam quality specifier for all modern clinical therapy accelerators? When can nonclinical beams in standards laboratories be used to calibrate ion chambers or measure kQ factors as a function of TPR10(20)? Based on detailed Monte Carlo simulations of Varian, Siemens, Elekta, and GE (Saturn) accelerators one can conclude that TPR10(20) is an adequate beam quality specifier for all these machines in the sense that for a given value of TPR10(20), the value of stopping-power ratios is the same. It is shown that, for the heavily filtered beams used in standards laboratories, TPR10(20) is an adequate beam quality specifier. It is also demonstrated that, for a larger range of bremsstrahlung beams than previously, %dd(10)x is a good beam quality specifier for all clinical beams as well as the lightly and heavily filtered beams in some standards laboratories. A criterion, based on the measured values of TPR10(20) and %dd(10)x for the beam, is proposed for determining whether a nonclinical beam is well specified by TPR10(20). Agreement between calculations for specific accelerators and measured beam quality specifiers is shown to be good, but agreement with published data for a variety of clinical accelerators is not as good. Possible reasons for the discrepancy are discussed. PMID- 12906173 TI - Effect of ethmoid sinus cavity on dose distribution at interface and how to correct for it: magnetic field with photon beams. AB - Recent work proposed the use of magnetic field as a solution to reduce the undesirable effect of air cavities on dose after the air/tissue interface. In contrast to the published work that looks into the problem with slab geometries, in this work we use actual anatomy based on CT images and the magnetic flux from a Helmholtz coil-pair configuration to investigate the problem and to evaluate the efficacy of the proposed solution. The EGS4 phantom was created using CT scans of the head at the level of the ethmoid sinus. The sinus measures 1.95 x 2.18 x 2.00 cm3. The grid size used is 0.15 x 0.15 x 0.4 cm3. Three different radiation beams, 1 x 1, 2 x 2, and 4 x 4 cm2, all 6 MV irradiate the phantom in two different configurations: single beam and parallel opposed. The magnetic field has three different strengths: 0.0, 0.5, and 1.0 T. These represent the maximum strength achieved in the middle of the configuration, between the two coils. The depth of the second buildup region in the absence of the magnetic field was used as the normalization point for the purpose of analysis. Dose was then scored at 0.23 cm after the air/tissue interface. A second phantom, very similar to the CT-based phantom, was created, but with the sinus cavity filled with unit-density tissue; everything else remained the same. This phantom provides a base to investigate the effect of the air cavity on dose. The phantom was termed the phantom without air, or PWA for short. We use the terms "dose reduction ratio" (DRR), defined as one minus the ratio of the dose in PWA to the dose with the presence of the cavity multiplied by 100% and the "dose improvement ratio" (DIR), defined as the ratio of dose with B to that without B, to evaluate the reduction in dose due to the cavity and the improvement in dose with magnetic field, respectively. For single beam geometry, the reduced dose ranged from 41% (1 x 1 cm2 beam) to less than 2% (4 x 4 cm2 beam). For the same single beam geometry, DIR ranged from 1.13 to 1.00 (DIR = 1 indicates no change) with 0.5 T, whereas it ranged from 1.44 to 1.05 for 1.0 T magnets. When an opposing beam was used, the reduced dose was not as severe, such that DRR ranged from 24% to less than 2%. Whereas the dose improvement ranged from 1.08 to 1.00 for 0.5 T, and from 1.23 to 1.01 for 1.0 T. PMID- 12906174 TI - The design of a new insert for calibration of LDR and HDR 192Ir sources in a well type ionization chamber. AB - Some well-type ionization chambers, present a very small sweet spot that are sufficient for small HDR sources. However, if a longer HDR source or LDR wires are calibrated, the positional uncertainty increases and an approximated correction factor must be applied, resulting in an increased uncertainty. One of the ways to avoid this problem would be to flatten the well chamber response by increasing its sweet spot region. This work uses the Monte Carlo code PENELOPE to simulate the response of a well-type chamber HDR-1000, with its original insert, by using an HDR 192Ir source and proposes a new insert design that increases its flatness region from 1.0 cm to approximately 4.0 cm (+/- 2.0 cm about the peak response). PMID- 12906175 TI - Reference dosimetry at the neutron capture therapy facility at Studsvik. AB - The purpose of this publication was to present and evaluate the methods for reference dosimetry in the epithermal neutron beam at the neutron capture therapy facility at Studsvik. Measurements were performed in a PMMA phantom and in air using ionization chambers and activation probes in order to calibrate the epithermal neutron beam. Appropriate beam-dependant calibration factors were determined using Monte Carlo methods for the detectors used in the present publication. Using the presented methodology, the photon, neutron and total absorbed dose to PMMA was determined with an estimated uncertainty of +/- 5.0%, +/- 25%, and +/- 5.5% (2 SD), respectively. The uncertainty of the determination of the photon absorbed dose was comparable to the case in conventional radiotherapy, while the uncertainty of the neutron absorbed dose is much higher using the present methods. The thermal neutron group fluence, i.e., the neutron fluence in the energy interval 0-0.414 eV, was determined with an estimated uncertainty of +/- 2.8% (2 SD), which is acceptable for dosimetry in epithermal neutron beams. PMID- 12906176 TI - Statistical analysis of decorrelation-based transducer tracking for three dimensional ultrasound. AB - The use of speckle decorrelation techniques to calculate the displacement of a moving transducer has shown promise. We describe a technique to estimate displacement between pairs of parallel planes without assuming that plane separation in the scan is uniform. We perform theoretical and empirical analyses of the bias and uncertainty in plane spacing estimates as a function of speckle size, patch size, and the number of planes used for normalization. Practically, only the central, linear region of the autocovariance curves can be used in this decorrelation method, which implies that distance between acquired image planes should be approximately half the speckle size. In this region, the uncertainty in estimated plane spacing was less than 15% for a 8.1 mm (axial) by 9.1 mm (lateral) patch and increased to 33% for an 8.1 mm (axial) by 1.5 mm (lateral) patch. The number of planes, Nz, used to calculate the normalization factors (averages of brightness and- squared brightness) was a major source of bias. Optimum Nz was found to be five to ten planes, depending on distance between acquired image planes, with a poor choice of Nz resulting in a bias of 10% or greater. A second source of bias is brightness gradients which, although they appear very slight on intensity images, can cause a large bias is the plane spacing estimates made using linearized data. PMID- 12906177 TI - Feature-guided analysis for reduction of false positives in CAD of polyps for computed tomographic colonography. AB - We evaluated the effect of our novel technique of feature-guided analysis of polyps on the reduction of false-positive (FP) findings generated by our computer aided diagnosis (CAD) scheme for the detection of polyps from computed tomography colonographic data sets. The detection performance obtained by use of feature guided analysis in the segmentation and feature analysis of polyp candidates was compared with that obtained by use of our previously employed fuzzy clustering technique. We also evaluated the effect of a feature called modified gradient concentration (MGC) on the detection performance. A total of 144 data sets, representing prone and supine views of 72 patients that included 14 patients with 21 colorectal polyps 5-25 mm in diameter, were used in the evaluation. At a 100% by-patient (95% by-polyp) detection sensitivity, the FP rate of our CAD scheme with feature-guided analysis based on round-robin evaluation was 1.3 (1.5) FP detections per patient. This corresponds to a 70-75% reduction in the number of FPs obtained by use of fuzzy clustering at the same sensitivity levels. Application of the MGC feature instead of our previously used gradient concentration feature did not improve the detection result. The results indicate that feature-guided analysis is useful for achieving high sensitivity and a low FP rate in our CAD scheme. PMID- 12906178 TI - Massive training artificial neural network (MTANN) for reduction of false positives in computerized detection of lung nodules in low-dose computed tomography. AB - In this study, we investigated a pattern-recognition technique based on an artificial neural network (ANN), which is called a massive training artificial neural network (MTANN), for reduction of false positives in computerized detection of lung nodules in low-dose computed tomography (CT) images. The MTANN consists of a modified multilayer ANN, which is capable of operating on image data directly. The MTANN is trained by use of a large number of subregions extracted from input images together with the teacher images containing the distribution for the "likelihood of being a nodule." The output image is obtained by scanning an input image with the MTANN. The distinction between a nodule and a non-nodule is made by use of a score which is defined from the output image of the trained MTANN. In order to eliminate various types of non-nodules, we extended the capability of a single MTANN, and developed a multiple MTANN (Multi MTANN). The Multi-MTANN consists of plural MTANNs that are arranged in parallel. Each MTANN is trained by using the same nodules, but with a different type of non nodule. Each MTANN acts as an expert for a specific type of non-nodule, e.g., five different MTANNs were trained to distinguish nodules from various-sized vessels; four other MTANNs were applied to eliminate some other opacities. The outputs of the MTANNs were combined by using the logical AND operation such that each of the trained MTANNs eliminated none of the nodules, but removed the specific type of non-nodule with which the MTANN was trained, and thus removed various types of non-nodules. The Multi-MTANN consisting of nine MTANNs was trained with 10 typical nodules and 10 non-nodules representing each of nine different non-nodule types (90 training non-nodules overall) in a training set. The trained Multi-MTANN was applied to the reduction of false positives reported by our current computerized scheme for lung nodule detection based on a database of 63 low-dose CT scans (1765 sections), which contained 71 confirmed nodules including 66 biopsy-confirmed primary cancers, from a lung cancer screening program. The Multi-MTANN was applied to 58 true positives (nodules from 54 patients) and 1726 false positives (non-nodules) reported by our current scheme in a validation test; these were different from the training set. The results indicated that 83% (1424/1726) of non-nodules were removed with a reduction of one true positive (nodule), i.e., a classification sensitivity of 98.3% (57 of 58 nodules). By using the Multi-MTANN, the false-positive rate of our current scheme was improved from 0.98 to 0.18 false positives per section (from 27.4 to 4.8 per patient) at an overall sensitivity of 80.3% (57/71). PMID- 12906179 TI - Dosimetric properties of an amorphous silicon electronic portal imaging device for verification of dynamic intensity modulated radiation therapy. AB - Dosimetric properties of an amorphous silicon electronic portal imaging device (EPID) for verification of dynamic intensity modulated radiation therapy (IMRT) delivery were investigated. The EPID was utilized with continuous frame-averaging during the beam delivery. Properties studied included effect of buildup, dose linearity, field size response, sampling of rapid multileaf collimator (MLC) leaf speeds, response to dose-rate fluctuations, memory effect, and reproducibility. The dependence of response on EPID calibration and a dead time in image frame acquisition occurring every 64 frames were measured. EPID measurements were also compared to ion chamber and film for open and wedged static fields and IMRT fields. The EPID was linear with dose and dose rate, and response to MLC leaf speeds up to 2.5 cm s(-1) was found to be linear. A field size dependent response of up to 5% relative to dmax ion-chamber measurement was found. Reproducibility was within 0.8% (1 standard deviation) for an IMRT delivery recorded at intervals over a period of one month. The dead time in frame acquisition resulted in errors in the EPID that increased with leaf speed and were over 20% for a 1 cm leaf gap moving at 1.0 cm s(-1). The EPID measurements were also found to depend on the input beam profile utilized for EPID flood-field calibration. The EPID shows promise as a device for verification of IMRT, the major limitation currently being due to dead-time in frame acquisition. PMID- 12906180 TI - Dose distributions in bifurcated coronary vessels treated with catheter-based photon and beta emitters in intravascular brachytherapy. AB - The dose distributions in the bifurcated vessels treated with intravascular brachytherapyline sources are complicated and depend on the bifurcation geometry consisting of a main and a branch vessel at different angles. To investigate the dosimetric effects at the bifurcation, calculations were performed on branching vessels of various bifurcation angles ranging from 20 degrees to 90 degrees. Two catheter based delivery systems were considered in the calculations using a 40 mm long radioactive sources of 192Ir or 90Sr/Y. It was assumed that the bifurcated vessel was treated in twoseparate source insertions, once for the main vessel and later for the branch vessel. Calculations were performed for different values of source gap from 0 to 9 mm, at the junction of main and branch vessels. Our results indicate that main vessel always receives a higher dose (up to 200%) when the branch vessel is also treated. Hot spots at portions of the main vessel near the junction cannot be totally avoided without severely underdosing the branch vessel. For bifurcation angle greater than 45 degrees, a 4 mm source gap can almost ensure that overdosing of the main vessel does not exceed 60% and underdosing of the branch vessel does not exceed 10% for 192Ir. However, for 90Sr/Y, the same is not possible unless the bifurcation angle is larger than 70 degrees. Dose heterogeneity using 90Sr/Y is more sensitive to the value of source gap than 192Ir because 90Sr/Y source provides a sharper dose-fall-off than 192Ir. For both photon and beta emitters, there is no acceptable solution for bifurcation angles less than 30 degrees, where the activity of the line source has a uniform distributions. Appropriate choice of gap at the junction can only help to reduce either overdosing of the main vessel or underdosing of the branch vessel, but not both. PMID- 12906181 TI - Testing and optimization of a semiautomatic prostate boundary segmentation algorithm using virtual operators. AB - Image analysis tasks such as size measurement and landmark-based registration require the user to select control points in an image. The output of such algorithms depends on the choice of control points. Since the choice of points varies from one user to the next, the requirement for user input introduces variability into the output of the algorithm. In order to test and/or optimize such algorithms, it is necessary to assess the multiplicity of outputs generated by the algorithm in response to a large set of inputs; however, the input of data requires substantial time and effort from multiple users. In this paper we describe a method to automate the testing and optimization of algorithms using "virtual operators," which consist of a set of spatial distributions describing how actual users select control points in an image. In order to construct the virtual operator, multiple users must repeatedly select control points in the image on which testing is to be performed. Once virtual operators are generated, control points for initializing the algorithm can be generated from them using a random number generator. Although an initial investment of time is required from the users in order to construct the virtual operator, testing and optimization of the algorithm can be done without further user interaction. We illustrate the construction and use of virtual operators by testing and optimizing our prostate boundary segmentation algorithm. The algorithm requires the user to select four control points on the prostate as input. PMID- 12906182 TI - Prostate boundary segmentation from 3D ultrasound images. AB - Segmenting, or outlining the prostate boundary is an important task in the management of patients with prostate cancer. In this paper, an algorithm is described for semiautomatic segmentation of the prostate from 3D ultrasound images. The algorithm uses model-based initialization and mesh refinement using an efficient deformable model. Initialization requires the user to select only six points from which the outline of the prostate is estimated using shape information. The estimated outline is then automatically deformed to better fit the prostate boundary. An editing tool allows the user to edit the boundary in problematic regions and then deform the model again to improve the final results. The algorithm requires less than 1 min on a Pentium III 400 MHz PC. The accuracy of the algorithm was assessed by comparing the algorithm results, obtained from both local and global analysis, to the manual segmentations on six prostates. The local difference was mapped on the surface of the algorithm boundary to produce a visual representation. Global error analysis showed that the average difference between manual and algorithm boundaries was -0.20 +/- 0.28 mm, the average absolute difference was 1.19 +/- 0.14 mm, the average maximum difference was 7.01 +/- 1.04 mm, and the average volume difference was 7.16% +/- 3.45%. Variability in manual and algorithm segmentation was also assessed: Visual representations of local variability were generated by mapping variability on the segmentation mesh. The mean variability in manual segmentation was 0.98 mm and in algorithm segmentation was 0.63 mm and the differences of about 51.5% of the points comprising the average algorithm boundary are insignificant (P < or = 0.01) to the manual average boundary. PMID- 12906183 TI - Particle selection for laser-accelerated proton therapy feasibility study. AB - In this paper we present calculations for the design of a particle selection system for laser-accelerated proton therapy. Laser-accelerated protons coming from a thin high-density foil have broad energy and angular spectra leading to dose distributions that cannot be directly used for therapeutic applications. Our solution to this problem is a compact particle selection and collimation device that delivers small pencil beams of protons with desired energy spectra. We propose a spectrometer-like particle selection and beam modulation system in which the magnetic field will be used to spread the protons spatially according to their energies and emitting angles. Subsequently, an aperture will be used to select the protons within a therapeutic window of energy (energy modulation). It will be shown that for the effective proton spatial differentiation, the primary collimation device should be used, which will collimate protons to the desired angular distribution and limit the spatial mixing of different energy protons once they have traveled through the magnetic system. Due to the angular proton distribution, the spatial mixing of protons of different energies will always be present and it will result in a proton energy spread with the width depending on the energy. For 250 MeV protons, the width (from the maximum to the minimum energy) is found to be 50 MeV for the magnetic field configuration used in our calculations. As the proton energy decreases, its energy width decreases as well, and for 80 MeV protons it equals 9 MeV. The presence of the energy width in the proton energy distribution will modify the depth dose curves needed for the energy modulation calculation. The matching magnetic field setup will ensure the refocusing of the selected protons and the final beam will be collimated by the secondary collimator. The calculations presented in this article show that the dose rate that the selection system can yield is on the order of D=260 Gy/min for a field size of 1 x 1 cm2. PMID- 12906184 TI - Incorporation of a laser range scanner into image-guided liver surgery: surface acquisition, registration, and tracking. AB - As image guided surgical procedures become increasingly diverse, there will be more scenarios where point-based fiducials cannot be accurately localized for registration and rigid body assumptions no longer hold. As a result, procedures will rely more frequently on anatomical surfaces for the basis of image alignment and will require intraoperative geometric data to measure and compensate for tissue deformation in the organ. In this paper we outline methods for which a laser range scanner may be used to accomplish these tasks intraoperatively. A laser range scanner based on the optical principle of triangulation acquires a dense set of three-dimensional point data in a very rapid, noncontact fashion. Phantom studies were performed to test the ability to link range scan data with traditional modes of image-guided surgery data through localization, registration, and tracking in physical space. The experiments demonstrate that the scanner is capable of localizing point-based fiducials to within 0.2 mm and capable of achieving point and surface based registrations with target registration error of less than 2.0 mm. Tracking points in physical space with the range scanning system yields an error of 1.4 +/- 0.8 mm. Surface deformation studies were performed with the range scanner in order to determine if this device was capable of acquiring enough information for compensation algorithms. In the surface deformation studies, the range scanner was able to detect changes in surface shape due to deformation comparable to those detected by tomographic image studies. Use of the range scanner has been approved for clinical trials, and an initial intraoperative range scan experiment is presented. In all of these studies, the primary source of error in range scan data is deterministically related to the position and orientation of the surface within the scanner's field of view. However, this systematic error can be corrected, allowing the range scanner to provide a rapid, robust method of acquiring anatomical surfaces intraoperatively. PMID- 12906185 TI - A dynamic approach to identifying desired physiological phases for cardiac imaging using multislice spiral CT. AB - In this investigation, we describe a quantitative technique to measure coronary motion, which can be correlated with cardiac image quality using multislice computed tomography (MSCT) scanners. MSCT scanners, with subsecond scanning, thin slice imaging (sub-millimeter) and volume scanning capabilities have paved the way for new clinical applications like noninvasive cardiac imaging. ECG-gated spiral CT using MSCT scanners has made it possible to scan the entire heart in a single breath-hold. The continuous data acquisition makes it possible for multiple phases to be reconstructed from a cardiac cycle. We measure the position and three-dimensional velocities of well-known landmarks along the proximal, mid, and distal regions of the major coronary arteries [left main (LM), left anterior descending (LAD), right coronary artery (RCA), and left circumflex (LCX)] during the cardiac cycle. A dynamic model (called the "delay algorithm") is described which enables us to capture the same physiological phase or "state" of the anatomy during the cardiac cycle as the instantaneous heart rate varies during the spiral scan. The coronary arteries are reconstructed from data obtained during different physiological cardiac phases and we correlate image quality of different parts of the coronary anatomy with phases at which minimum velocities occur. The motion characteristics varied depending on the artery, with the highest motion being observed for RCA. The phases with the lowest mean velocities provided the best visualization. Though more than one phase of relative minimum velocity was observed for each artery, the most consistent image quality was observed during mid-diastole ("diastasis") of the cardiac cycle and was judged to be superior to other reconstructed phases in 92% of the cases. In the process, we also investigated correlation between cardiac arterial states and other measures of motion, such as the left ventricular volume during a cardiac cycle, which earlier has been demonstrated as an example of how anatomic-specific information can be used in a knowledge-based cardiac CT algorithm. Using these estimates in characterizing cardiac motion also provides realistic simulation models for higher heart rates and also in optimizing volume reconstructions for individual segments of the cardiac anatomy. PMID- 12906186 TI - Flat panel detector-based cone beam computed tomography with a circle-plus-two arcs data acquisition orbit: preliminary phantom study. AB - Cone beam computed tomography (CBCT) has been investigated in the past two decades due to its potential advantages over a fan beam CT. These advantages include (a) great improvement in data acquisition efficiency, spatial resolution, and spatial resolution uniformity, (b) substantially better utilization of x-ray photons generated by the x-ray tube compared to a fan beam CT, and (c) significant advancement in clinical three-dimensional (3D) CT applications. However, most studies of CBCT in the past are focused on cone beam data acquisition theories and reconstruction algorithms. The recent development of x ray flat panel detectors (FPD) has made CBCT imaging feasible and practical. This paper reports a newly built flat panel detector-based CBCT prototype scanner and presents the results of the preliminary evaluation of the prototype through a phantom study. The prototype consisted of an x-ray tube, a flat panel detector, a GE 8800 CT gantry, a patient table and a computer system. The prototype was constructed by modifying a GE 8800 CT gantry such that both a single-circle cone beam acquisition orbit and a circle-plus-two-arcs orbit can be achieved. With a circle-plus-two-arcs orbit, a complete set of cone beam projection data can be obtained, consisting of a set of circle projections and a set of arc projections. Using the prototype scanner, the set of circle projections were acquired by rotating the x-ray tube and the FPD together on the gantry, and the set of arc projections were obtained by tilting the gantry while the x-ray tube and detector were at the 12 and 6 o'clock positions, respectively. A filtered backprojection exact cone beam reconstruction algorithm based on a circle-plus-two-arcs orbit was used for cone beam reconstruction from both the circle and arc projections. The system was first characterized in terms of the linearity and dynamic range of the detector. Then the uniformity, spatial resolution and low contrast resolution were assessed using different phantoms mainly in the central plane of the cone beam reconstruction. Finally, the reconstruction accuracy of using the circle plus-two-arcs orbit and its related filtered backprojection cone beam volume CT reconstruction algorithm was evaluated with a specially designed disk phantom. The results obtained using the new cone beam acquisition orbit and the related reconstruction algorithm were compared to those obtained using a single-circle cone beam geometry and Feldkamp's algorithm in terms of reconstruction accuracy. The results of the study demonstrate that the circle-plus-two-arcs cone beam orbit is achievable in practice. Also, the reconstruction accuracy of cone beam reconstruction is significantly improved with the circle-plus-two-arcs orbit and its related exact CB-FPB algorithm, as compared to using a single circle cone beam orbit and Feldkamp's algorithm. PMID- 12906187 TI - Ionization chamber volume averaging effects in dynamic intensity modulated radiation therapy beams. AB - The commercial cylindrical ionization chamber ionization integration accuracy of dynamically moving fields was evaluated. The ionization chambers were exposed to long (14 cm), narrow (0.6, 1.0, 2.0, and 4.0 cm) 6 MV and 18 MV fields. Rather than rely on the linear accelerator to reproducibly scan across the chamber, the chambers were scanned beneath fixed portals. A water-equivalent phantom was constructed with cavities that matched the chambers and placed on a computer controlled one-dimensional table. Computer-controlled electrometers were utilized in continuous charge integrate mode, with 10 samples of the charge, along with time stamps, acquired for each chamber location. A reference chamber was placed just beneath the linear accelerator jaws to adjust for variations in linear accelerator dose rate. The scan spatial resolution was selected to adequately sample regions of steep dose gradient and second spatial derivative (curvature). A fixed measurement in a 10 x 10 cm2 field was used to normalize the profiles to absolute chamber response. Three ionization chambers were tested, a microchamber (0.009 cm3), a Farmer chamber (0.6 cm3) and a waterproof scanning chamber (0.125 cm3). The larger chambers exhibited severe under-response at the small field's centers, but all of the chambers, independent of orientation, accurately integrated the ionization across the scanned portal. This indicates that the tested ionization chambers provide accurate integrated charges in regions of homogeneous dose regions. Partial integration (less than the field width plus the chamber length plus 2 cm), yielded integration errors of greater than 1% and 2% for 6 MV and 18 MV, respectively, with errors for the Farmer chamber of greater than 10% even for the 4 cm wide field. PMID- 12906188 TI - The value of scatter removal by a grid in full field digital mammography. AB - Our objective in this study was to investigate the usefulness of an anti-scatter grid in digital mammography using a contrast detail phantom. The mammography system we investigated was a GE Senographe 2000D. We carried out phantom measurements under various conditions with and without using the anti-scatter grid. A new version of the CDMAM phantom (version 3.4) was used. This phantom consists of a matrix of square cells with disks of varying size and contrast. For given exposure conditions detectability of these disks can be determined and used for construction of contrast detail curves. Previously, a computer program was developed at our institute that performs a fully automatic analysis of the phantom recordings using the ideal observer model. Breast thickness was simulated by a homogeneous layer of PMMA in the range of 1 to 7 cm. Series of images were recorded for different KeV and target-filter combinations depending on the simulated thickness. The dose was kept constant for each thickness with and without using a grid. It appeared that image quality improved for simulated breast thickness below 5 cm when the grid was removed. In the range from 5 to 7 cm contrast detail curves obtained with or without a grid were similar. Results suggest that for compressed breast thickness in the range of 1 to 7 cm a grid might not be needed in the digital mammography system we investigated. Below 5 cm, omitting the grid may allow lower dose to the patient without losing image quality. PMID- 12906189 TI - On site evaluation of three flat panel detectors for digital radiography. AB - During a tender we evaluated the image performance of three commercially available active matrix flat panel imagers (AMFPI) for general radiography, one based on direct detection method (Se photoconductor) the other two on indirect detection method (CsI phosphor). Basic image quality parameters (MTF, NNPS, DQE) were evaluated with particular attention to dose and energy dependence. As it is known, presampling modulation transfer function (MTF) of selenium based detector is very high (at 70 kV, 2 cycles/mm, 2.5 microGy, about 0.80). Indirect detection panels exhibit a comparable (lower) resolution (at 70 kV, 2 cycles/mm, 2.5 microGy, MTF is about 0.34 for both the systems analyzed) and a more pronounced energy and dose dependence could also be noted in one of them. As a consequence of the very high resolution, the normalized noise power spectrum (NNPS) of the direct system is substantially flat, very similar to a white noise. Considering that the sensitive layer of all detectors is the same (0.5 mm), the relatively higher NNPS values are related to selenium absorption properties (lower Z respect to CsI:Tl) and detector inherent noise. NNPSs of the other systems, at low frequencies, are comparable but the frequency dependence is significantly different. At 70 kV, 2.5 microGy, 0.5 cycles/mm detective quantum efficiency (DQE) is about 0.35 for the direct detection system, and about the same (0.6) for the indirect ones. The combined effect of additive and multiplicative noise components makes DQE dependence on dose not monotonic. DQE present a maximum for an intermediate exposure. This complex behavior may be useful to characterize the systems in terms of the monodimensional integral over the frequency of DQE (IDQE). Both visual contrast-detail experiment and the direct evaluation of the signal-to-noise ratio confirmed, at least in a qualitative way, the system performances predicted by IDQE. PMID- 12906190 TI - Thermoluminescent dosimetry of the SourceTech Medical model STM1251 125I seed. AB - Many new models of 125I seeds are being introduced, mainly due to the increase in prostate seed implants. We have evaluated the SourceTech Medical (STM), model STM1251, 125I seed using thermoluminescent dosimeters (TLDs) in a solid water phantom. TLD cubes, LiF TLD-100, with dimension 1 mm on each edge, were irradiated at various distances, 1, 2, 3, and 5 cm, at angles ranging from 0 degrees to 90 degrees in 10 degrees increments. Sensitivity calibration of the TLDs was achieved by irradiation to 10 cGy with 6 MV x rays from a clinical linear accelerator, Clinac 600C. Concurrent with the 125I seed exposures, several TLDs were also exposed to 10 cGy with the 600C as a control set. Dose rates per unit air kerma strength were determined based on the 1999 NIST traceable standard for the STM1251 seed. They are presented as a function of distance r and angle theta. The TG-43 parameters, including the dose rate constant, lambda, anisotropy function, F(r,theta), radial dose function, g(r), anisotropy factor, phian(r), and anisotropy constant, phi, were obtained for use in radiation treatment planning software. The value of lambda was determined as 1.07 +/- 5.5% cGy U(-1) h(-1), which is comparable to model 6702 and to the value determined using the point extrapolation method by Kirov and Williamson. We also find agreement between our TLD data and their Monte Carlo results for g(r), F(r,theta), phian(r), and phi. Additionally, agreement is found with the TLD data of Li and Williamson for lambda and g(r). PMID- 12906191 TI - The effects of intra-fraction organ motion on the delivery of intensity-modulated field with a multileaf collimator. AB - Intensity-modulated radiation therapy can be conveniently delivered with a multileaf collimator. With this method, the entire field is not delivered at once, but rather it is composed of many subfields defined by the leaf positions as a function of beam on time. At any given instant, only these subfields are delivered. During treatment, if the organ moves, part of the volume may move in or out of these subfields. Due to this interplay between organ motion and leaf motion the delivered dose may be different from what was planned. In this work, we present a method that calculates the effects of organ motion on delivered dose. The direction of organ motion may be parallel or perpendicular to the leaf motion, and the effect can be calculated for a single fraction or for multiple fractions. Three breast patients and four lung patients were included in this study,with the amplitude of the organ motion varying from +/- 3.5 mm to +/- 10 mm, and the period varying from 4 to 8 seconds. Calculations were made for these patients with and without organ motion, and results were examined in terms of isodose distribution and dose volume histograms. Each calculation was repeated ten times in order to estimate the statistical uncertainties. For selected patients, calculations were also made with conventional treatment technique. The effects of organ motion on conventional techniques were compared relative to that on IMRT techniques. For breast treatment, the effect of organ motion primarily broadened the penumbra at the posterior field edge. The dose in the rest of the treatment volume was not significantly affected. For lung treatment, the effect also broadened the penumbra and degraded the coverage of the planning target volume (PTV). However, the coverage of the clinical target volume (CTV) was not much affected, provided the PTV margin was adequate. The same effects were observed for both IMRT and conventional treatment techniques. For the IMRT technique, the standard deviations of ten samples of a 30-fraction calculation were very small for all patients, implying that over a typical treatment course of 30 fractions, the delivered dose was very close to the expected value. Hence, under typical clinical conditions, the effect of organ motion on delivered dose can be calculated without considering the interplay between the organ motion and the leaf motion. It can be calculated as the weighted average of the dose distribution without organ motion with the distribution of organ motion. Since the effects of organ motion on dose were comparable for both IMRT and conventional techniques, the PTV margin should remain the same for both techniques. PMID- 12906192 TI - Image quality in two phosphor-based flat panel digital radiographic detectors. AB - Two general types of phosphor screens are currently used in indirect digital radiographic systems: structured phosphor screens and turbid phosphor screens. The purpose of the study was to experimentally compare the image quality characteristics of two flat-panel digital radiography detectors with similar electronics and pixel sizes (0.127 mm), but otherwise equipped with the two types of screens (0.6-mm-thick structured CsI and Lanex Regular). The presampled modulation transfer functions (MTFs) of the detectors were assessed using an edge method. The noise power spectra (NPS) were measured by two-dimensional Fourier analysis of uniformly-exposed radiographs at 50-100 kVp with 19 mm added Al filtration. The detective quantum efficiencies (DQEs) were assessed from the MTF, the NPS, and estimates of the ideal signal-to-noise ratio. The MTF measures of the two detectors were generally similar above a spatial frequency of 2 mm(-1), with approximately 2.5 and approximately 3.8 mm(-1) spatial frequencies corresponding to 0.2 MTF and 0.1 MTF, respectively. Below 2 mm(-1), the MTF for the CsI-based detector was slightly higher by an average of 0.07. At 70 kVp, the measured DQE values in the diagonal (and axial) direction(s) at spatial frequencies of 0.15 mm(-1) and 2.5 mm(-1) were 78% (78%) and 26% (20%) for the CsI-based detector, and 20% (20%) and 7% (6%) for the Lanex-based detector, respectively. The comparative findings experimentally confirm that in indirect flat-panel detectors, structured phosphor screens provide a more favorable tradeoff between resolution and noise compared to turbid-phosphor screens, effectively increasing the detection efficiency of the detector without a negative impact on the detector's spatial resolution response. PMID- 12906193 TI - Segmental and dynamic intensity-modulated radiotherapy delivery techniques for micro-multileaf collimator. AB - A leaf sequencing algorithm has been implemented to deliver segmental and dynamic multileaf collimated intensity-modulated radiotherapy (SMLC-IMRT and DMLC-IMRT, respectively) using a linear accelerator equipped with a micro-multileaf collimator (mMLC). The implementation extends a previously published algorithm for the SMLC-IMRT to include the dynamic MLC-IMRT method and several dosimetric considerations. The algorithm has been extended to account for the transmitted radiation and minimize the leakage between opposing and neighboring leaves. The underdosage problem associated with the tongue-and-groove design of the MLC is significantly reduced by synchronizing the MLC leaf movements. The workings of the leaf sequencing parameters have been investigated and the results of the planar dosimetric investigations show that the sequencing parameters affect the measured dose distributions as intended. Investigations of clinical cases suggest that SMLC and DMLC delivery methods produce comparable results with leaf sequences obtained by root-mean-square (RMS) errors specification of 1.5% and lower, approximately corresponding to 20 or more segments. For SMLC-IMRT, there is little to be gained by using an RMS error specification smaller than 2%, approximately corresponding to 15 segments; however, more segments directly translate to longer treatment time and more strain on the MLC. The implemented leaf synchronization method does not increase the required monitor units while it reduces the measured TG underdoses from a maximum of 12% to a maximum of 3% observed with single field measurements of representative clinical cases studied. PMID- 12906194 TI - Measurement of focal spot size with slit camera using computed radiography and flat-panel based digital detectors. AB - The purpose of this study was to evaluate the use of digital x-ray imaging detectors for the measurement of diagnostic x-ray tube focal spot size using a slit camera. Slit camera images of two focal spots for a radiographic x-ray tube were acquired with direct-exposure film (DF) (as specified by the National Electrical Manufacturers Association [NEMA] Standards Publication No. XR 5, 1992), computed radiography (CR) imaging plates, and an a-Si:H/CsI:Tl-based flat panel (FP) detector. Images obtained with the CR and the FP were acquired over a broad range of detector entrance exposure levels. The DF slit images were evaluated according to NEMA specifications (visually, using a 7x magnifying glass with reticule) by six medical physicists. Additionally, the DF images were digitized and the focal spot sizes obtained from the digital profiles of the slit. The CR and the FP images were analyzed in a manner similar to the digitized DF images. It took less than 20 minutes for a complete CR or FP measurement of focal spot size in two dimensions. In comparison, a typical DF measurement with visual evaluation takes at least 60 minutes, in our experience. In addition to a great reduction in measurement time achieved by using digital detectors, the tube loading requirements were reduced to approximately 20 mAs compared with approximately 1000 mAs when using the DF technique. The calculated focal spot sizes for CR and FP differed from those of digitized DF by -2.4% to +4.8% (sigma=2.5%), far less than the -16.6% to +9.3% (sigma=8.1%) variability introduced by the visual evaluation of the slit image. In addition, the calculated focal spot sizes for the CR and the FP images maintained a coefficient of variation <1.0% over the broad range of exposure levels. Based upon these results, we conclude that (1) FP and CR detectors yield consistent results in measurements of x-ray tube focal spot sizes, (2) compared to DF, CR and FP significantly reduce measurement time and tube loading requirements, (3) CR and FP readily permit digital profile analysis, thereby eliminating observer error, and (4) unlike DF, CR and FP are independent of exposure level. PMID- 12906195 TI - A fluence convolution method to account for respiratory motion in three dimensional dose calculations of the liver: a Monte Carlo study. AB - We describe the implementation of a fluence convolution method to account for the influence of superior-inferior (SI) respiratory induced motion on a Monte Carlo based dose calculation of a tumor located in the liver. This method involves convolving the static fluence map with a function describing the SI motion of the liver-the motion function has been previously derived from measurements of diaphragm movement observed under fluoroscopy. Significant differences are noted between fluence-convolved and static dose distributions in an example clinical treatment plan; hot and cold spots (on the order of 25%) are observed in the fluence-convolved plan at the superior and inferior borders of the liver, respectively. This study illustrates that the fluence convolution method can be incorporated into Monte Carlo dose calculation algorithms to account for some of the effects of patient breathing during radiotherapy treatment planning, thus leading to more accurate dose calculations. PMID- 12906196 TI - Computer aided detection of masses in mammography using subregion Hotelling observers. AB - We propose to investigate the use of the subregion Hotelling observer for the basis of a computer aided detection scheme for masses in mammography. A database of 1320 regions of interest (ROIs) was selected from the DDSM database collected by the University of South Florida using the Lumisys scanner cases. The breakdown of the cases was as follows: 656 normal ROIs, 307 benign ROIs, and 357 cancer ROIs. Each ROI was extracted at a size of 1024 x 1024 pixels and sub-sampled to 128 x 128 pixels. For the detection task, cancer and benign cases were considered positive and normal was considered negative. All positive cases had the lesion centered in the ROI. We chose to investigate the subregion Hotelling observer as a classifier to detect masses. The Hotelling observer incorporates information about the signal, the background, and the noise correlation for prediction of positive and negative and is the optimal detector when these are known. For our study, 225 subregion Hotelling observers were set up in a 15 x 15 grid across the center of the ROIs. Each separate observer was designed to "observe," or discriminate, an 8 x 8 pixel area of the image. A leave one out training and testing methodology was used to generate 225 "features," where each feature is the output of the individual observers. The 225 features derived from separate Hotelling observers were then narrowed down by using forward searching linear discriminants (LDs). The reduced set of features was then analyzed using an additional LD with receiver operating characteristic (ROC) analysis. The 225 Hotelling observer features were searched by the forward searching LD, which selected a subset of 37 features. This subset of 37 features was then analyzed using an additional LD, which gave a ROC area under the curve of 0.9412 +/- 0.006 and a partial area of 0.6728. Additionally, at 98% sensitivity the overall classifier had a specificity of 55.9% and a positive predictive value of 69.3%. Preliminary results suggest that using subregion Hotelling observers in combination with LDs can provide a strong backbone for a CAD scheme to help radiologists with detection. Such a system could be used in conjunction with CAD systems for false positive reduction. PMID- 12906197 TI - Elimination of importance factors for clinically accurate selection of beam orientations, beam weights and wedge angles in conformal radiation therapy. AB - A method of simultaneously optimizing beam orientations, beam weights, and wedge angles for conformal radiotherapy is presented. This method removes the need for importance factors by optimizing one objective only, subject to a set of rigid constraints. This facilitates the production of inverse solutions which, without trial-and-error modification of importance factors, precisely satisfy the specified constraints. The algorithm minimizes an objective function which is based upon the single objective to be optimized, but which is forced to an artificially high value when the constraints are not met, so that only satisfactory solutions are allowed. Due to the complex nature of the objective function space, including multiple local minima separated by large regions of plateau, a random search technique equivalent to fast simulated annealing is used for producing inverse plans. To illustrate the novel features of the new algorithm, a simulation is first presented, for the case of a cylindrical phantom. The morphology of the objective function space is shown to be significantly different for the new algorithm, compared to that for a conventional quadratic objective function. Clinical cases for prostate and craniopharyngioma are then presented. For the prostate case, the objective is to reduce irradiated rectal volume. Three-field, four-field, and six-field optimizations, with or without orientation optimization, are shown to provide solutions which are consistent with previously reported plans and class solutions. For the craniopharyngioma case, which involves the use of a high precision stereotactic conformal technique, the objective is to reduce the irradiated volume of normal brain. Practically feasible beam angles are produced which, compared to a standard plan, provide a small but worthwhile sparing of normal brain. The algorithm is thereby shown to be robust and suitable for clinical application. PMID- 12906198 TI - Integrated density of a lesion: a quantitative, mammographically derived, invariable measure. AB - A method for quantitatively estimating lesion "size" from mammographic images was developed and evaluated. The main idea behind the measure, termed "integrated density" (ID), is that the total x-ray attenuation attributable to an object is theoretically invariant with respect to the projected view and object deformation. Because it is possible to estimate x-ray attenuation of a lesion from relative film densities, after appropriate corrections for background, the invariant property of the measure is expected to result in an objective method for evaluating the "sizes" of breast lesions. ID was calculated as the integral of the estimated image density attributable to a lesion, relative to surrounding background, over the area of the lesion and after corrections for the nonlinearity of the film characteristic curve. This effectively provides a measure proportional to lesion volume. We computed ID and more traditional measures of size (such as "mass diameter" and "effective size") for 100 pairs of ipsilateral mammographic views, each containing a lesion that was relatively visible in both views. The correlation between values calculated for each measure from corresponding pairs of ipsilateral views were computed and compared. All three size-related measures (mass diameter, effective size, and ID) exhibited reasonable linear relationship between paired views (r2>0.7, P<0.001). Specifically, the ID measures for the 100 masses were found to be highly correlated (r2=0.9, P<0.001) between ipsilateral views of the same mass. The correlation increased substantially (r2=0.95), when a measure with linear dimensions of length was defined as the cube root of ID. There is a high degree of correlation between ID-based measures obtained from different views of the same mass. ID-based measures showed a higher degree of invariance than mass diameter or effective size. PMID- 12906199 TI - Dose distribution close to metal implants in Gamma Knife Radiosurgery: a Monte Carlo study. AB - Materials with high atomic numbers favor the occurrence of the photoelectric effect when they are irradiated with gamma rays. Therefore, the photoelectric effects of metal implants within the target regions in Gamma Knife Radiosurgery are worth studying. In the present work, Monte Carlo simulations using EGS4 were employed to investigate the resulting dose enhancements. A dose enhancement as high as 10% was observed close to a platinum implant along the x and y axes, while no significant dose enhancements were observed for silver, stainless steel 301, and titanium ones. A dose enhancement as high as 20% was observed close to the platinum implant along the z axis at the superior position of the metal phantom interface and was 10% higher for other metal implants. PMID- 12906201 TI - Robustness and precision of an automatic marker detection algorithm for online prostate daily targeting using a standard V-EPID. AB - An algorithm for the daily localization of the prostate using implanted markers and a standard video-based electronic portal imaging device (V-EPID) has been tested. Prior to planning, three gold markers were implanted in the prostate of seven patients. The clinical images were acquired with a BeamViewPlus 2.1 V-EPID for each field during the normal course radiotherapy treatment and are used off line to determine the ability of the automatic marker detection algorithm to adequately and consistently detect the markers. Clinical images were obtained with various dose levels from ranging 2.5 to 75 MU. The algorithm is based on marker attenuation characterization in the portal image and spatial distribution. A total of 1182 clinical images were taken. The results show an average efficiency of 93% for the markers detected individually and 85% for the group of markers. This algorithm accomplishes the detection and validation in 0.20-0.40 s. When the center of mass of the group of implanted markers is used, then all displacements can be corrected to within 1.0 mm in 84% of the cases and within 1.5 mm in 97% of cases. The standard video-based EPID tested provides excellent marker detection capability even with low dose levels. The V-EPID can be used successfully with radiopaque markers and the automatic detection algorithm to track and correct the daily setup deviations due to organ motions. PMID- 12906200 TI - Compensator quality control with an amorphous silicon EPID. AB - The calibration and quality control of compensators is conventionally performed with an ion chamber in a water-equivalent phantom. In our center, the compensator factor and four off-axis fluence ratios are measured to verify the central axis beam modulation and orientation of the compensator. Here we report the investigation of an alternative technique for compensator quality control using an amorphous silicon electronic portal imaging device (a-Si EPID). Preliminary experiments were performed to identify appropriate EPID operating parameters for this relative dosimetric study and also to quantify EPID operation. The pixel value versus energy fluence response of the EPID for both open and compensated fields was then determined, and expressed via calibration curves. For open fields the response was seen to be linear, whereas for compensated fields it exhibited a small quadratic component. To account for field size effects, we measured EPID scatter factors. These exhibited small but non-negligible dependencies on compensator thickness and source-detector distance. Finally, a number of test and clinical compensators were evaluated to assess the suitability of the EPID for compensator quality control. Our results indicate that the a-Si EPID can measure clinical compensator factors and off-axis energy fluence ratios to within 2% of values measured by a Farmer chamber on average, and so is a suitable ion chamber replacement. PMID- 12906202 TI - Computerized analysis of shadowing on breast ultrasound for improved lesion detection. AB - Sonography is being considered for the screening of women at high risk for breast cancer. We are developing computerized detection methods to aid in the localization of lesions on breast ultrasound images. The detection scheme presented here is based on the analysis of posterior acoustic shadowing, since posterior acoustic shadowing is observed for many malignant lesions. The method uses a nonlinear filtering technique based on the skewness of the gray level distribution within a kernel of image data. The database used in this study included 400 breast ultrasound cases (757 images) consisting of complicated cysts, solid benign lesions, and malignant lesions. At a false-positive rate of 0.25 false positives per image, a detection sensitivity of 80% by case (66% by image) was achieved for malignant lesions. The performance for the overall database (at 0.25 false positives per image) was less at 42% sensitivity by case (30% by image) due to the more limited presence of posterior acoustic shadowing for benign solid lesions and the presence of posterior acoustic enhancement for cysts. Our computerized method for the detection of lesion shadows alerts radiologists to lesions that exhibit posterior acoustic shadowing. While this is not a characterization method, its performance is best for lesions that exhibit posterior acoustic shadowing such as malignant and, to a lesser extent, benign solid lesions. This method, in combination with other computerized sonographic detection methods, may ultimately help facilitate the use of ultrasound for breast cancer screening. PMID- 12906203 TI - Performance evaluation of a "dual-side read" dedicated mammography computed radiography system. AB - The image quality of a dedicated mammography computed radiography (CR) system was characterized. A unique feature of this system is that it collects image signals from both sides of the storage phosphor. Measurements of the modulation transfer function (MTF), noise power spectrum (NPS), and detective quantum efficiency (DQE) were made. This work included improvements in our measurement methods to specifically account for the detrimental effects of system glare on the MTF and to accurately characterize the low-frequency NPS components. Image quality measurements were performed using a 25 kVp beam filtered with 2 mm Al and an exposure range of 1 to 100 mR (87 to 870 microGy). The DQE was found to decrease with increasing exposure due to an increased contribution of storage phosphor structure noise. The DQE of this system was compared to similar measurements made using a standard CR system. The dual-side read system demonstrated superior DQE compared to the standard system. The decrease in DQE with increasing exposure was more severe for the standard system than the dual-side read system. This finding suggests that the CR system noise was reduced for the dual-side read system compared to the standard system. PMID- 12906205 TI - X-ray scattering in full-field digital mammography. AB - Effects of x-ray scattering on full-field digital mammography are analyzed with the scattering model of Seibert and Boone [Med. Phys. 15, 567-575 (1988)]. A new method is introduced for the estimation of model parameters from measurements. It is shown that with breasts thinner than a certain threshold, removing the anti scatter grid leads to an improved contrast-to-noise ratio with a smaller patient dose. A fast approximate algorithm is presented for determining the scattered field in a gridless digital mammogram. PMID- 12906204 TI - Effect of applicator diameter on lesion size from high temperature interstitial ultrasound thermal therapy. AB - High temperature ultrasound thermal therapy using interstitial and external approaches is becoming increasingly acceptable as a minimally invasive clinical treatment for cancerous and benign disease. The diameter of an interstitial applicator can influence its clinical practicality and effectiveness as well as application site. The purpose of this study was to determine whether the use of larger ultrasound transducers and the inherent increase in applicator size could be justified by potentially producing larger lesion diameters. Four applicator configurations and sizes were studied using ex vivo tissue experiments in liver and beef and using acoustic and biothermal simulations. Catheter-cooled and internally cooled applicators with outer diameters between 2.2 and 4.0 mm produced 3.5 to 5.0 cm diameter lesions in ex vivo liver and 3.0 to 3.5 cm lesions in ex vivo beef muscle with 20-40 W/cm applied for 10 min. Larger applicators produced lesions with radial penetration depths superior to their smaller counterparts at power levels in the 20-40 W/cm range. The higher cooling rates along the outer surface of the larger diameter applicators due to their greater surface area was a dominant factor in increasing lesion size. The higher cooling rates pushed the maximum temperature farther from the applicator surface and reduced the formation of high acoustic attenuation tissue zones. Applicator configuration and frequency (6.7-8.2 MHz) had less influence on lesion size than diameter in the ranges studied. Acoustic and biothermal simulations matched the experimental data well and were applied to model these applicators within sites of clinical interest such as prostate, uterine fibroid, brain, and normal liver. Lesions of 3.9 to 4.7 cm diameter were predicted for moderately perfused tissues such as prostate and fibroid and 2.8 to 3.2 cm for highly perfused tissues such as normal liver. In sites such as uterine fibroid where larger applicators placed using an endoscopic approach could be tolerated, treatment volume increases of 37% were predicted for an applicator diameter increase from 2.4 to 4.0 mm. PMID- 12906206 TI - System performance of a prototype flat-panel imager operated under mammographic conditions. AB - The results of an empirical and theoretical investigation of the performance of a high-resolution, active matrix flat-panel imager performed under mammographic conditions are reported. The imager is based upon a prototype, indirect detection active matrix array incorporating a discrete photodiode in each pixel and a pixel to-pixel pitch of 97 microm. The investigation involved three imager configurations corresponding to the use of three different x-ray converters with the array. The converters were a conventional Gd2O2S-based mammographic phosphor screen (Min-R) and two structured CsI:Tl scintillators: one optimized for high spatial resolution (FOS-HR) and the other for high light output (FOS-HL). Detective quantum efficiency for mammographic exposures ranging from approximately 2 to approximately 40 mR at 26 kVp were determined for each imager configuration through measurements of x-ray sensitivity, modulation transfer function (MTF), and noise power spectrum (NPS). All configurations were found to provide significant presampling MTF at frequencies beyond the Nyquist frequency of the array, approximately 5.2 mm(-1) , consistent with the high spatial resolution of the converters. In addition, the effect of additive electronic noise on the NPS was found to be significantly larger for the configuration with lower system gain (FOS-HR) than for the configurations with higher gain (Min-R, FOS-HL). The maximum DQE values obtained with the CsI:Tl scintillators were considerably greater than those obtained with the Min-R screen due to the significantly lower Swank noise of the scintillators. Moreover, DQE performance was found to degrade with decreasing exposure, although this exposure-dependence was considerably reduced for the higher gain configurations. Theoretical calculations based on the cascaded systems model were found to be in generally good agreement with these empirically determined NPS and DQE values. In this study, we provide an example of how cascaded systems modeling can be used to identify factors limiting system performance and to examine trade-offs between factors toward the goal of maximizing performance. PMID- 12906207 TI - Generalized monitor unit calculation for the Varian enhanced dynamic wedge field. AB - The generalized monitor unit (MU) calculation equation for the Varian enhanced dynamic wedge (EDW) is derived. The assumption of this MU calculation method is that the wedge factor of the EDW at the center of the field is a function of field size, the position of the center of the field in the wedge direction, and the final position of the moving jaw. The wedge factors at the center of the field in both symmetric and asymmetric fields are examined. The difference between calculated and measured wedge factors is within 1.0%. The method developed here is easy to implement. The only datum required in addition to the standard set of conventional physical wedge implementation data is the off-axis output factor for the open field in the reference condition. The off-center point calculation is also examined. For the off-center point calculation, the dose profile in the wedge direction for the largest EDW field is used to obtain the relative off-center ratio in any smaller wedge field. The accuracy of the off center point calculation decreases when the point of calculation is too close to the field edge. PMID- 12906208 TI - A systematic study of imaging uncertainties and their impact on 125I prostate brachytherapy dose evaluation. AB - In order to calculate the dose distribution delivered by a prostate brachytherapy implant, the seed positions and prostate volume are normally identified on post implant CT images. We have systematically considered the impact of uncertainties in contouring the prostate, seed localization, and visualization of all the seeds on the calculated dose distributions, dose-volume histograms, and predicted radiobiological outcome. This study was done for a collection of 27 clinical 125I prostate brachytherapy implants, performed at the London Regional Cancer Centre during our early adoption of this technique. For these clinical dose distributions, the median D90 was 76% of the prescription dose, or 110 Gy, and the median V90 was 80%. We calculated the changes in these dosimetric indices (D90 and V90) and radiobiological outcome (SF2 TCP) as a function of contouring uncertainty, seed localization uncertainty, inability to localize all of the seeds, and binary combinations of these three. The results are presented for a range of uncertainties, which allows the possible application of these results to a variety of imaging modalities that have differing spatial resolutions. We found that both contouring uncertainties and seed localization uncertainties had a large impact on the predicted radiobiological outcome, but that seed localization uncertainties of 6 mm had the largest impact on the dosimetric indices. We also found that the variability in both the predicted radiobiological and dosimetric outcome was largest for contouring uncertainties of 4-8 mm. We conclude that accounting for contouring uncertainties is crucial in accurately deducing the DVHs for post-implant prostate brachytherapy, and hence enabling valid correlation with ultimate clinical outcome. PMID- 12906209 TI - Comparison of IMRT optimization based on a pencil beam and a superposition algorithm. AB - To investigate the role of sophisticated dose calculation methods for treatment planning, we compared conventional pencil beam optimized 6 and 15 MV intensity modulated treatment plans with optimizations based on the superposition technique. Five lung and five head and neck IMRT cases with spatial resolutions of bixels and dose voxels usually employed in clinical practice were considered for tumor volumes between 15 and 500 cm3. We investigated the systematic error of the pencil beam algorithm and the pencil beam induced error to the optimal solution of bixel weights. For the lung cases, the pencil beam overestimated the mean dose deposited inside the planning target volume (PTV) by about 8%, for small lung tumors even up to 20.6%. In the head and neck cases only a slight overestimation in mean PTV dose of 1.5% was observed. The optimization with the superposition method substantially improved the dose coverage of the considered radiation targets. Additionally, for the head and neck cases, the brainstem was significantly spared by about 4% mean PTV dose through the use of the superposition technique. Our studies showed that, in target regions with intricate tissue inhomogeneities, superposition or Monte Carlo techniques have to be used for the optimization and the final dose calculation of intensity modulated treatment plans. PMID- 12906210 TI - Mammogram synthesis using a three-dimensional simulation. III. Modeling and evaluation of the breast ductal network. AB - A method is proposed for realistic simulation of the breast ductal network as part of a computer three-dimensional (3-D) breast phantom. The ductal network is simulated using tree models. Synthetic trees are generated based upon a description of ductal branching by ramification matrices (R matrices), whose elements represent the probabilities of branching at various levels of a tree. We simulated the ductal network of the breast, consisting of multiple lobes, by random binary trees (RBT). Each lobe extends from the ampulla and consists of branching ductal segments of decreasing size, and the associated terminal ductal lobular units. The lobes follow curved paths that project from the nipple toward the chest wall. We have evaluated the RBT model by comparing manually-traced ductal networks from 25 projections of ductal lobes in clinical galactograms and manually-traced networks from 23 projections of synthetic RBTs. A root-mean square (rms) fractional error of 41%, between the R-matrix elements corresponding to clinical and synthetic images, was computed. This difference was influenced by projection and segmentation artifacts and by the limited number of available images. In addition, we analyzed 23 synthetic trees generated using R matrices computed from clinical images. A comparison of these synthetic and clinical images yielded a rms fractional error of 11%, suggesting the possibility that a more appropriate model of the ductal branching morphology may be developed. Rejection of the RBT model also suggests the existence of a relationship between ductal branching morphology and the state of mammary developmentand pathology. PMID- 12906211 TI - Test of GEANT3 and GEANT4 nuclear models for 160 MeV protons stopping in CH2. AB - Monte Carlo simulations are used for many problems in proton radiation therapy, some of which are sensitive to the nuclear interaction model. The available models have been little tested in the regime of interest, namely in their ability to predict the secondary particle yield, including their angle and energy, when 70-250 MeV protons stop in various materials. The present study provides one such test in carbon, complementing a previous one in copper. Using a multilayer Faraday cup we have measured the projected range distribution of charged nuclear secondaries from 160 MeV protons stopping in polyethylene (CH2). To test the popular GEANT Monte Carlo we have simulated the experiment with GEANT3 using the "Gheisha" (default) and "Fluka" models and with GEANT4.5 using the "low-energy" and "precompound" models. The GEANT3/Fluka and GEANT4/precompound simulations agree moderately well with the observed range distribution. The data are given in a convenient form for testing other Monte Carlo programs. PMID- 12906212 TI - Comment on "A Monte Carlo study of IMRT beamlets in inhomogeneous media". PMID- 12906213 TI - McDonald's takes antimicrobial stance. PMID- 12906214 TI - First member of the equine family cloned. PMID- 12906215 TI - Illinois legislature passes controversial dangerous-dog bill. PMID- 12906216 TI - More on animal-friendly farming. PMID- 12906217 TI - Recognizing signs of pain. PMID- 12906218 TI - Continued debate on admission policies. PMID- 12906219 TI - Differing opinions on regulating CAVM standards. PMID- 12906220 TI - Differing opinions on regulating CAVM standards. PMID- 12906221 TI - Wants to clear up Internet misinformation regarding aspartame. PMID- 12906222 TI - What is your diagnosis? Avulsion and dorsal displacement of greater trochanter and fractures of the acetabulum and left pecten of the pubis. PMID- 12906223 TI - Reversing the downward trend. PMID- 12906224 TI - Evaluation of risk factors for bite wounds inflicted on caregivers by dogs and cats in a veterinary teaching hospital. AB - OBJECTIVE: To identify factors associated with increased risk of being bitten by a dog or cat in a veterinary teaching hospital. DESIGN: Unmatched case-control study. STUDY POPULATION: 207 animal caregivers. PROCEDURE: Case subjects (n = 75) were any caregiver that reported being bitten by a dog or cat. Control subjects (n = 132) were randomly selected from a list of all caregivers interacting with dogs or cats. Information on the characteristics of the caregivers, characteristics of the dogs and cats, and the nature of the interaction between the dog or cat and the caregiver was obtained by use of self-administered questionnaires. RESULTS: Caregivers were more likely to be bitten by dogs or cats that had warning signs on their cages indicating the potential to bite or that were considered difficult to handle. Caregivers interacting with cats or with older dogs and cats were more likely to be bitten. Only 37 to 55% of dogs and cats that had characteristics traditionally associated with biting or were considered likely to bite were muzzled. CONCLUSIONS AND CLINICAL RELEVANCE: Muzzling dogs and cats should be considered more frequently. Dogs and cats considered to have the propensity to bite frequently do bite, and precautions, such as muzzling, should be taken if the medical condition or conformation of the dog or cat is amenable to this type of restraint. PMID- 12906225 TI - Diagnostic quality of percutaneous kidney biopsy specimens obtained with laparoscopy versus ultrasound guidance in dogs. AB - OBJECTIVE: To compare diagnostic quality of percutaneous kidney biopsy specimens obtained with laparoscopy versus ultrasound guidance in dogs and compare diagnostic quality of specimens obtained with 14- versus 18-gauge biopsy needles. DESIGN: Prospective study. ANIMALS: 10 healthy dogs. PROCEDURE: In each dog, 2 biopsy specimens were obtained from each kidney, 1 with a 14-gauge biopsy needle and 1 with an 18-gauge biopsy needle. Biopsy specimens were obtained from 1 kidney by means of ultrasound guidance and from the contralateral kidney by means of direct viewing during laparoscopy. Number of glomeruli, quality of the biopsy specimen, proportion of specimens that contained muscle tissue, and proportion of specimens with fragmentation or crushing were determined. RESULTS: Mean +/- SD number of glomeruli (32.6 +/- 11.0) in laparoscopic, 14-gauge biopsy specimens was significantly higher than mean number of glomeruli in ultrasound-guided, 14 gauge specimens; mean number of glomeruli in ultrasound-guided, 18-gauge specimens; and mean number of glomeruli in laparoscopic, 18-gauge specimens. All 10 laparoscopic, 14-gauge biopsy specimens were classified as excellent. The proportion of 18-gauge biopsy specimens with crushing or fragmentation was significantly higher than the proportion of 14-gauge specimens. One of the kidneys biopsied with ultrasound guidance had a large amount of hemorrhage. Hemorrhage was modest and transient following laparoscopic biopsy. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that excellent-quality renal biopsy specimens with large numbers of glomeruli can be obtained with 14-gauge, double spring-activated biopsy needles during laparoscopy. Renal biopsy specimens obtained with 18-gauge biopsy needles frequently had few glomeruli and often were crushed or fragmented, increasing the difficulty in making an accurate diagnosis. PMID- 12906226 TI - Efficacy and safety of topical administration of selamectin for treatment of ear mite infestation in rabbits. AB - OBJECTIVE: To evaluate the efficacy and safety of topical administration of selamectin in rabbits naturally infested with Psoroptes cuniculi. DESIGN: Randomized controlled trial. ANIMALS: 48 mixed-breed domestic rabbits with active P. cuniculi mite populations and clinical ear lesions. PROCEDURES: Rabbits were randomly allocated to 1 of 6 treatment groups. On day 0, rabbits in groups 1 and 2 were given vehicle, rabbits in groups 3 and 4 were given selamectin at a dose of 6 mg/kg (2.7 mg/lb), and rabbits in groups 5 and 6 were given selamectin at a dose of 18 mg/kg (8.2 mg/lb). On day 28, rabbits in groups 2, 4, and 6 were given a second dose of vehicle or selamectin. Otoscopic examinations were performed and ear lesion size was measured weekly for 8 weeks. Quantitative viable mite counts were performed on day 56. RESULTS: On days 7 through 56, lesion sizes for all selamectin-treated groups were significantly lower than sizes for control groups; there were no significant differences in lesion sizes among selamectin-treated groups. All rabbits in the 2 control groups had viable adult P. cuniculi mites for the duration of the study, as determined by otoscopic examination, whereas all rabbits in the 4 selamectin-treated groups were free from P. cuniculi mites on days 7 through 56. No adverse reactions associated with selamectin treatment were observed. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that topical application of selamectin at a dose of 6 or 18 mg/kg can completely eliminate mites from rabbits naturally infested with P. cuniculi. PMID- 12906227 TI - Clinical use of low-profile cystostomy tubes in four dogs and a cat. AB - Traditional cystostomy tubes (used for temporary or permanent diversion of urine in dogs and cats) are long (> or = 22 cm) and cumbersome to stabilize, requiring sutures or bandages to hold the tube against the body. Use of a low-profile gastrostomy port system as a low-profile cystostomy tube (LPCT) in 4 dogs and a cat was investigated; owner satisfaction with the device was assessed. Technical difficulty associated with placement and management of LPCTs was similar to that for traditional cystostomy tubes; with LPCTs, activity and mobility of pets was not compromised, and bandaging was not required. Complications included lower urinary tract infection, mild peristomal leakage of urine and leakage from components of the system, and subcutaneous peristomal infection. Four of 5 owners considered the tube to be easy to use; all owners said they would be comfortable repeating their decision to use the LPCT in their pet. PMID- 12906228 TI - Use of a plate-rod construct and principles of biological osteosynthesis for repair of diaphyseal fractures in dogs and cats: 47 cases (1994-2001). AB - OBJECTIVE: To determine outcome for dogs and cats with diaphyseal fractures in which a plate-rod construct was used for fracture repair. DESIGN: Retrospective study. ANIMALS: 35 dogs and 12 cats. PROCEDURES: Medical records and radiographs were reviewed to obtain information concerning signalment, fracture severity, construct design, time to radiographic union, complications, and outcome. Clients were contacted by telephone to obtain information on complications, limb usage, and overall satisfaction with the procedure. RESULTS: 31 femoral, 9 humeral, and 7 tibial fractures were assessed. Thirteen fractures consisted of 2 fragments, 22 consisted of 3 to 5 fragments, and 12 consisted of > 5 fragments. Forty-six of 47 (98%) fractures reached union. Mean +/- SD times to radiographic union were 7.5 +/- 2.7 weeks for the dogs and 4.8 +/- 1.3 weeks for the cats. There were 4 short term, minor complications and 15 long-term complications (2 major and 13 minor). Owners of 21 of 30 dogs (70%) and 9 of 12 cats reported that their animals had normal limb function. Twenty-six of 28 dog owners (93%) and 12 of 12 cat owners indicated that they were satisfied with results of the procedure. As surgery time increased, time to union also increased. Time to union for fractures with > 5 fragments was significantly shorter than time to union for fractures with < or = 5 fragments. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that plate-rod constructs can successfully be used for repair of diaphyseal fractures of a wide range of severity in dogs and cats. PMID- 12906229 TI - Odds of moderate or severe gastric ulceration in racehorses receiving antiulcer medications. AB - OBJECTIVE: To determine the odds of moderate or severe gastric ulceration in racehorses treated with various antiulcer medications. DESIGN: Unmatched case control study. ANIMALS: 798 horses in active race training (252 Thoroughbreds and 546 Standardbreds). Only horses that had been receiving a single antiulcer medication or no antiulcer medication for at least 2 weeks prior to examination were included. PROCEDURE: Gastroscopy was performed on each horse by a single individual who was not aware of the horses' antiulcer treatments, and severity of gastric ulceration was scored. Signalment and medication history were recorded. Logistic regression was used to determine whether identification of moderate or severe ulceration was associated with treatment, age, breed, or sex. Treatments were grouped as no treatment, buffer, sucralfate, histamine type 2 receptor antagonist, compounded omeprazole, proprietary omeprazole at a low dosage, and proprietary omeprazole at a high dosage. RESULTS: Only proprietary omeprazole was associated with significantly lower odds of moderate or severe ulceration, compared with no treatment. Risks of moderate or severe gastric ulceration in horses receiving a buffer, sucralfate, a histamine type 2 receptor antagonist, or compounded omeprazole were not significantly different from risks in horses receiving no antiulcer medication. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that the proprietary formulation of omeprazole was associated with a significantly lower risk of moderate or severe gastric ulceration, compared with no treatment, in racehorses in active race training, whereas other antiulcer medications were not. PMID- 12906230 TI - Thyroid function in mature horses ingesting endophyte-infected fescue seed. AB - OBJECTIVE: To determine whether ingestion of fescue seed infected with the endophyte Neotyphodium coenophialum would alter thyroid function in adult horses. DESIGN: Original study. ANIMALS: 4 adult mares that were not pregnant and 6 adult geldings. PROCEDURE: Thyrotropin releasing hormone stimulation tests were performed while horses received a standard diet and after infected seed (2.3 kg/d [5 lb/d]) had been fed for 1 and 2 months. Serum prolactin concentrations were measured to verify endophyte absorption. RESULTS: Serum prolactin concentrations indicated that at least 8 of 10 horses absorbed the endophyte. Baseline concentrations of thyroid stimulating hormone, total and free triiodothyronine, and total and free thyroxine and the change in hormone concentrations in response to administration of thyrotropin releasing hormone (1 mg, i.v.) were not altered by ingestion of endophyte-infected fescue seed. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that ingestion of fescue seed infected with the endophytic fungus N. coenophialum for 2 months has little effect on thyroid function in adult horses that are not pregnant. PMID- 12906231 TI - Economic implications of bovine leukemia virus infection in mid-Atlantic dairy herds. AB - OBJECTIVE: To determine the baseline costs of bovine leukemia virus (BLV) infection, including costs of clinical disease and subclinical infection, in a dairy herd representative of the mid-Atlantic region and compare these costs with the cost of a test-and-manage BLV control program. DESIGN: Stochastic spreadsheet model. SAMPLE POPULATION: A commercial Holstein dairy herd with 100 milking cows. PROCEDURES: A spreadsheet model was developed. The overall cost of infection included the cost of clinical disease (ie, lymphosarcoma [LS]) and the effects of subclinical infection on milk production and premature culling. Model input values and distributions were designed to reflect economic conditions in the mid Atlantic region. Relative costs of infection and control were calculated for infection prevalences of 20, 50, and 80%. RESULTS: Estimated mean cost to the producer per case of LS was 412 dollars; for a herd with a 50% prevalence of BLV infection, annual incidence of LS was 0.66. Mean annual cost of subclinical infection at a 50% prevalence of infection was 6,406 dollars. Mean annual cost of a test-and-manage control program was 1,765 dollars. The cost of clinical disease and subclinical infection varied substantially with the prevalence of infection, whereas the cost of control varied with herd size. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that a basic BLV control program may be economically beneficial in herds in which the prevalence of BLV infection is > or = 12.5%. Farm-specific considerations may factor prominently when weighing the costs and benefits of an individual BLV control program. PMID- 12906232 TI - Use of a supraplastron approach to the coelomic cavity for repair of an esophageal tear in a loggerhead sea turtle. AB - A juvenile loggerhead sea turtle (Caretta caretta) was referred for removal of a fishhook lodged in the coelomic portion of the esophagus. Attempts at manual and endoscopic extraction were unsuccessful and resulted in a 6-cm tear in the coelomic portion of the esophagus. A supraplastron approach was made to the coelomic cavity, allowing access to and repair of the esophageal tear. The turtle recovered well and was able to be released to the wild. The surgical approach was associated with less trauma and shorter healing and rehabilitation times, compared with traditional plastron osteotomy. PMID- 12906233 TI - A normal distribution for tensor-valued random variables: applications to diffusion tensor MRI. AB - Diffusion tensor magnetic resonance imaging (DT-MRI) provides a statistical estimate of a symmetric, second-order diffusion tensor of water, D, in each voxel within an imaging volume. We propose a new normal distribution, p(D) alpha exp( 1/2 D: A: D), which describes the variability of D in an ideal DT-MRI experiment. The scalar invariant, D : A : D, is the contraction of a positive definite symmetric, fourth-order precision tensor, A, and D. A correspondence is established between D: A: D and the elastic strain energy density function in continuum mechanics--specifically between D and the second-order infinitesimal strain tensor, and between A and the fourth-order tensor of elastic coefficients. We show that A can be further classified according to different classical elastic symmetries (i.e., isotropy, transverse isotropy, orthotropy, planar symmetry, and anisotropy). When A is an isotropic fourth-order tensor, we derive an explicit analytic expression for p(D), and for the distribution of the three eigenvalues of D, p(gamma1, gamma2, gamma3), which are confirmed by Monte Carlo simulations. We show how A can be estimated from either real or synthetic DT-MRI data for any given experimental design. Here we propose a new criterion for an optimal experimental design: that A be an isotropic fourth-order tensor. This condition ensures that the statistical properties of D (and quantities derived from it) are rotationally invariant. We also investigate the degree of isotropy of several DT MRI experimental designs. Finally, we show that the univariate and multivariate distributions are special cases of the more general tensor-variate normal distribution, and suggest how to generalize p(D) to treat normal random tensor variables that are of third- (or higher) order. We expect that this new distribution, p(D), should be useful in feature extraction; in developing a hypothesis testing framework for segmenting and classifying noisy, discrete tensor data; and in designing experiments to measure tensor quantities. PMID- 12906234 TI - Activation detection in fMRI using a maximum energy ratio statistic obtained by adaptive spatial filtering. AB - An adaptive spatial filtering method is proposed that takes into account contextual information in fMRI activation detection. This filter replaces the time series of each voxel with a weighted average of time series of a small neighborhood around it. The filter coefficients at each voxel are derived so as to maximize a test statistic designed to indicate the presence of activation. This statistic is the ratio of the energy of the filtered time series in a signal subspace to the energy of the residuals. It is shown that the filter coefficients and the maximum energy ratio can be found through a generalized eigenproblem. This approach equates the filter coefficients to the elements of an eigenvector corresponding to the largest eigenvalue of a specific matrix, while the largest eigenvalue itself becomes the maximum energy ratio that can be used as a statistic for detecting activation. The distribution of this statistic under the null hypothesis is derived by a nonparametric permutation technique in the wavelet domain. Also, in this paper we introduce a new set of basis vectors that define the signal subspace. The space spanned by these basis vectors covers a wide range of possible hemodynamic response functions (HRF) and is applicable to both event related and block design fMRI signal analysis. This approach circumvents the need for a priori assumptions about the exact shape of the HRF. Resting-state experimental fMRI data were used to assess the specificity of the method, showing that the actual false-alarm rate of the proposed method is equal or less than its expected value. Analysis of simulated data and motor task fMRI datasets from six volunteers using the method proposed here showed an improved sensitivity as compared to a conventional test with a similar statistic applied to spatially smoothed data. PMID- 12906235 TI - Combined MR data acquisition of multicontrast images using variable acquisition parameters and K-space data sharing. AB - A new technique to reduce clinical magnetic resonance imaging (MRI) scan time by varying acquisition parameters and sharing k-space data between images, is proposed. To improve data utilization, acquisition of multiple images of different contrast is combined into a single scan, with variable acquisition parameters including repetition time (TR), echo time (TE), and echo train length (ETL). This approach is thus referred to as a "combo acquisition." As a proof of concept, simulations of MRI experiments using spin echo (SE) and fast SE (FSE) sequences were performed based on Bloch equations. Predicted scan time reductions of 25%-50% were achieved for 2-contrast and 3-contrast combo acquisitions. Artifacts caused by nonuniform k-space data weighting were suppressed through semi-empirical optimization of parameter variation schemes and the phase encoding order. Optimization was assessed by minimizing three quantitative criteria: energy of the "residue point spread function (PSF)," energy of "residue profiles" across sharp tissue boundaries, and energy of "residue images." In addition, results were further evaluated by quantitatively analyzing the preservation of contrast, the PSF, and the signal-to-noise ratio. Finally, conspicuity of lesions was investigated for combo acquisitions in comparison with standard scans. Implications and challenges for the practical use of combo acquisitions are discussed. PMID- 12906236 TI - Fluorescence-enhanced optical tomography using referenced measurements of heterogeneous media. AB - A three-dimensional image reconstruction for fluorescence-enhanced frequency domain photon migration (FDPM) measurements in turbid media is developed and investigated for three different simulated measurement types: 1) absolute emission measurement, or emission measurements of phase and amplitude attenuation made for a given incident point source of excitation light; 2) referenced emission measurements made relative to an excitation measurement conducted at a single reference point away from the incident source; and 3) referenced emission measurements made relative to the excitation measurement conducted at identical points of detection. The image reconstruction algorithm employs a gradient-based constrained truncated Newton (CONTN) method which implements a bounding parameter, which can be used to govern the level of contrast used to discriminate tissue volumes from heterogeneous background tissues. Reverse differentiation technique is used to calculate the gradients. Using simulated data with superimposed noise to achieve a signal-to-noise ratio of 55 and 35 dB to mimic experimental excitation and emission FDPM measurements, respectively, we show the robustness of emission measurements referenced to excitation light. We investigate the performance of algorithm CONTN using these measurement techniques and show that the absorption coefficients due to fluorophore are reconstructed by CONTN accurately and efficiently. Furthermore, we demonstrate the performance of the bounding parameter for rejection of background artifacts owing to background tissue heterogeneity. PMID- 12906237 TI - Blind deblurring of spiral CT images. AB - To discriminate fine anatomical features in the inner ear, it has been desirable that spiral computed tomography (CT) may perform beyond their current resolution limits with the aid of digital image processing techniques. In this paper, we develop a blind deblurring approach to enhance image resolution retrospectively without complete knowledge of the underlying point spread function (PSF). An oblique CT image can be approximated as the convolution of an isotropic Gaussian PSF and the actual cross section. Practically, the parameter of the PSF is often unavailable. Hence, estimation of the parameter for the underlying PSF is crucially important for blind image deblurring. Based on the iterative deblurring theory, we formulate an edge-to-noise ratio (ENR) to characterize the image quality change due to deblurring. Our blind deblurring algorithm estimates the parameter of the PSF by maximizing the ENR, and deblurs images. In the phantom studies, the blind deblurring algorithm reduces image blurring by about 24%, according to our blurring residual measure. Also, the blind deblurring algorithm works well in patient studies. After fully automatic blind deblurring, the conspicuity of the submillimeter features of the cochlea is substantially improved. PMID- 12906238 TI - Spatial domain filtering for fast modification of the tradeoff between image sharpness and pixel noise in computed tomography. AB - In computed tomography (CT), selection of a convolution kernel determines the tradeoff between image sharpness and pixel noise. For certain clinical applications it is desirable to have two or more sets of images with different settings. So far, this typically requires reconstruction of several sets of images. We present an alternative approach using default reconstruction of sharp images and online filtering in the spatial domain allowing modification of the sharpness-noise tradeoff in real time. A suitable smoothing filter function in the frequency domain is the ratio of smooth and original (sharp) kernel. Efficient implementation can be achieved by a Fourier transform of this ratio to the spatial domain. Separating the two-dimensional spatial filtering into two subsequent one-dimensional filtering stages in the x and y directions using a Gaussian approximation for the convolution kernel further reduces computational complexity. Due to efficient implementation, interactive modification of the filter settings becomes possible, which can completely replace the variety of different reconstruction kernels. PMID- 12906240 TI - Elastic registration in the presence of intensity variations. AB - We have developed a general-purpose registration algorithm for medical images and volumes. This method models the transformation between images as locally affine but globally smooth. The model also explicitly accounts for local and global variations in image intensities. This approach is built upon a differential multiscale framework, allowing us to capture both large- and small-scale transformations. We show that this approach is highly effective across a broad range of synthetic and clinical medical images. PMID- 12906239 TI - Interpolation artifacts in multimodality image registration based on maximization of mutual information. AB - Mutual information (MI) is an increasingly popular match metric for multimodality image registration. However, its value is affected by interpolation, which may limit registration accuracy. The purpose of this study was to characterize the artifacts from eight interpolators and to investigate efficient strategies to overcome these artifacts. The interpolators were: 1) nearest neighbor; 2) linear; 3) cubic Catmull-Rom; 4) Hamming-windowed sinc; 5) partial volume; 6) NN with jittered sampling (JIT); 7) NN with histogram blurring (BLUR); and 8) NN with JIT and BLUR. The impact of interpolation on MI was evaluated in two dimensions over different translational and rotational misregistration. Interpolation caused spurious fluctuations in MI whenever the voxel grids had coinciding periodicities and were nearly aligned. The artifacts did not lessen by using intensity interpolators with wider support (e.g., cubic Catmull-Rom, Hamming-windowed sinc). PV could lead to either arch artifacts or inverted-arch artifacts, depending on the relative voxel sizes. Several strategies reduced artifacts and improved registration robustness: JIT, BLUR, avoiding an extreme number of intensity bins, and resampling the images in a rotated orientation with different relative voxel sizes (e.g., pi/3). These findings also apply to related methods, including normalized MI, joint entropy, and Hill's third moment. PMID- 12906241 TI - Isosurfaces as deformable models for magnetic resonance angiography. AB - Vascular disease produces changes in lumenal shape evident in magnetic resonance angiography (MRA). However, quantification of vascular shape from MRA is problematic due to image artifacts. Prior deformable models for vascular surface reconstruction primarily resolve problems of initialization of the surface mesh. However, initialization can be obtained in a trivial manner for MRA using isosurfaces. We propose a methodology for deforming the isosurface to conform to the boundaries of objects in the image with minimal a priori assumptions of object shape. As in conventional methods, external forces attract the surface toward edges in the image. However, smoothing is produced by a moment that aligns the normals of adjacent surface triangles. Notably, the moment produces no translational motion of surface triangles. The deformable isosurface was applied to a digital phantom of a stenotic artery, to MRA of three renal arteries with atherosclerotic disease and MRA of one carotid artery with atherosclerotic disease. Results of the surface reconstruction from the deformable model were compared with conventional X-ray angiography for the renal arteries. Measurement of the degree of stenosis of the renal arteries was within 12% +/- 6%. The deformable model provided improvements over the isosurface in all cases in terms of measurement of the degree of stenosis or improving the surface smoothness. PMID- 12906242 TI - Intensity-based segmentation of microarray images. AB - The underlying principle in microarray image analysis is that the spot intensity is a measure of the gene expression. This implicitly assumes the gene expression of a spot to be governed entirely by the distribution of the pixel intensities. Thus, a segmentation technique based on the distribution of the pixel intensities is appropriate for the current problem. In this paper, clustering-based segmentation is described to extract the target intensity of the spots. The approximate boundaries of the spots in the microarray are determined by manual adjustment of rectilinear grids. The distribution of the pixel intensity in a grid containing a spot is assumed to be the superposition of the foreground and the local background. The k-means clustering technique and the partitioning around medoids (PAM) were used to generate a binary partition of the pixel intensity distribution. The median (k-means) and the medoid (PAM) of the cluster members are chosen as the cluster representatives. The effectiveness of the clustering-based segmentation techniques was tested on publicly available arrays generated in a lipid metabolism experiment (Callow et al., 2000). The results are compared against those obtained using the region-growing approach (SPOT) (Yang et al., 2001). The effect of additive white Gaussian noise is also investigated. PMID- 12906243 TI - A finite-element approach for Young's modulus reconstruction. AB - Modulus imaging has great potential in soft-tissue characterization since it reveals intrinsic mechanical properties. A novel Young's modulus reconstruction algorithm that is based on finite-element analysis is reported here. This new method overcomes some limitations in other Young's modulus reconstruction methods. Specifically, it relaxes the force boundary condition requirements so that only the force distribution at the compression surface is necessary, thus making the new method more practical. The validity of the new method is demonstrated and the performance of the algorithm with noise in the input data is tested using numerical simulations. Details of how to apply this method under clinical conditions is also discussed. PMID- 12906244 TI - Texture-based classification of atherosclerotic carotid plaques. AB - There are indications that the morphology of atherosclerotic carotid plaques, obtained by high-resolution ultrasound imaging, has prognostic implications. The objective of this study was to develop a computer-aided system that will facilitate the characterization of carotid plaques for the identification of individuals with asymptomatic carotid stenosis at risk of stroke. A total of 230 plaque images were collected which were classified into two types: symptomatic because of ipsilateral hemispheric symptoms, or asymptomatic because they were not connected with ipsilateral hemispheric events. Ten different texture feature sets were extracted from the manually segmented plaque images using the following algorithms: first-order statistics, spatial gray level dependence matrices, gray level difference statistics, neighborhood gray tone difference matrix, statistical feature matrix, Laws texture energy measures, fractal dimension texture analysis, Fourier power spectrum and shape parameters. For the classification task a modular neural network composed of self-organizing map (SOM) classifiers, and combining techniques based on a confidence measure were used. Combining the classification results of the ten SOM classifiers inputted with the ten feature sets improved the classification rate of the individual classifiers, reaching an average diagnostic yield (DY) of 73.1%. The same modular system was implemented using the statistical k-nearest neighbor (KNN) classifier. The combined DY for the KNN system was 68.8%. The results of this paper show that it is possible to identify a group of patients at risk of stroke based on texture features extracted from ultrasound images of carotid plaques. This group of patients may benefit from a carotid endarterectomy whereas other patients may be spared from an unnecessary operation. PMID- 12906246 TI - Single-shot MR imaging using trapezoidal-gradient-based Lissajous trajectories. AB - A novel single-shot trapezoidal-gradient-based Lissajous trajectory is described and implemented on a 3-tesla magnetic resonance (MR) scanner. A feature of this trajectory is that its sampling points are located on a nonequidistant rectangular grid, which permits the usage of one-dimensional optimal algorithms to increase the robustness and speed of image reconstruction. Another advantage of the trajectory is that two images with different effective echo times can be obtained within a single excitation, which might be used for fast T2* mapping, in functional MR imaging scanning of brain activity associated with mental processes. Potential artifacts in reconstructed images were investigated and methods for suppressing these artifacts were developed. Experiments on normal subjects at rest and during brain activation were performed to demonstrate the feasibility of the new sequence. PMID- 12906245 TI - Uncertainty in the output of artificial neural networks. AB - Analysis of the performance of artificial neural networks (ANNs) is usually based on aggregate results on a population of cases. In this paper, we analyze ANN output corresponding to the individual case. We show variability in the outputs of multiple ANNs that are trained and "optimized" from a common set of training cases. We predict this variability from a theoretical standpoint on the basis that multiple ANNs can be optimized to achieve similar overall performance on a population of cases, but produce different outputs for the same individual case because the ANNs use different weights. We use simulations to show that the average standard deviation in the ANN output can be two orders of magnitude higher than the standard deviation in the ANN overall performance measured by the Az value. We further show this variability using an example in mammography where the ANNs are used to classify clustered microcalcifications as malignant or benign based on image features extracted from mammograms. This variability in the ANN output is generally not recognized because a trained individual ANN becomes a deterministic model. Recognition of this variability and the deterministic view of the ANN present a fundamental contradiction. The implication of this variability to the classification task warrants additional study. PMID- 12906247 TI - Analysis of event-related fMRI data using best clustering bases. AB - We explore a new paradigm for the analysis of event-related functional magnetic resonance images (fMRI) of brain activity. We regard the fMRI data as a very large set of time series x(i) (t), indexed by the position i of a voxel inside the brain. The decision that a voxel i(o) is activated is based not solely on the value of the fMRI signal at i(o), but rather on the comparison of all time series x(i) (t) in a small neighborhood Wi(o) around i(o). We construct basis functions on which the projection of the fMRI data reveals the organization of the time series x(i) (t) into activated and nonactivated clusters. These clustering basis functions are selected from large libraries of wavelet packets according to their ability to separate the fMRI time series into the activated cluster and a nonactivated cluster. This principle exploits the intrinsic spatial correlation that is present in the data. The construction of the clustering basis functions described in this paper is applicable to a large category of problems where time series are indexed by a spatial variable. PMID- 12906248 TI - Segmentation and analysis of the human airway tree from three-dimensional X-ray CT images. AB - The lungs exchange air with the external environment via the pulmonary airways. Computed tomography (CT) scanning can be used to obtain detailed images of the pulmonary anatomy, including the airways. These images have been used to measure airway geometry, study airway reactivity, and guide surgical interventions. Prior to these applications, airway segmentation can be used to identify the airway lumen in the CT images. Airway tree segmentation can be performed manually by an image analyst, but the complexity of the tree makes manual segmentation tedious and extremely time-consuming. We describe a fully automatic technique for segmenting the airway tree in three-dimensional (3-D) CT images of the thorax. We use grayscale morphological reconstruction to identify candidate airways on CT slices and then reconstruct a connected 3-D airway tree. After segmentation, we estimate airway branchpoints based on connectivity changes in the reconstructed tree. Compared to manual analysis on 3-mm-thick electron-beam CT images, the automatic approach has an overall airway branch detection sensitivity of approximately 73%. PMID- 12906249 TI - Locating the optic nerve in a retinal image using the fuzzy convergence of the blood vessels. AB - We describe an automated method to locate the optic nerve in images of the ocular fundus. Our method uses a novel algorithm we call fuzzy convergence to determine the origination of the blood vessel network. We evaluate our method using 31 images of healthy retinas and 50 images of diseased retinas, containing such diverse symptoms as tortuous vessels, choroidal neovascularization, and hemorrhages that completely obscure the actual nerve. On this difficult data set, our method achieved 89% correct detection. We also compare our method against three simpler methods, demonstrating the performance improvement. All our images and data are freely available for other researchers to use in evaluating related methods. PMID- 12906250 TI - A new algorithm for border description of polarized light surface microscopic images of pigmented skin lesions. AB - The aim of this study was to provide mathematical descriptors for the border of pigmented skin lesion images and to assess their efficacy for distinction among different lesion groups. New descriptors such as lesion slope and lesion slope regularity are introduced and mathematically defined. A new algorithm based on the Catmull-Rom spline method and the computation of the gray-level gradient of points extracted by interpolation of normal direction on spline points was employed. The efficacy of these new descriptors was tested on a data set of 510 pigmented skin lesions, composed by 85 melanomas and 425 nevi, by employing statistical methods for discrimination between the two populations. PMID- 12906251 TI - Guide-wire tracking during endovascular interventions. AB - A method is presented to extract and track the position of a guide wire during endovascular interventions under X-ray fluoroscopy. The method can be used to improve guide-wire visualization in low-quality fluoroscopic images and to estimate the position of the guide wire in world coordinates. A two-step procedure is utilized to track the guide wire in subsequent frames. First, a rough estimate of the displacement is obtained using a template-matching procedure. Subsequently, the position of the guide wire is determined by fitting a spline to a feature image. The feature images that have been considered enhance line-like structures on: 1) the original images; 2) subtraction images; and 3) preprocessed images in which coherent structures are enhanced. In the optimization step, the influence of the scale at which the feature is calculated and the additional value of using directional information is investigated. The method is evaluated on 267 frames from ten clinical image sequences. Using the automatic method, the guide wire could be tracked in 96% of the frames, with a similar accuracy to three observers, although the position of the tip was estimated less accurately. PMID- 12906253 TI - Mutual-information-based registration of medical images: a survey. AB - An overview is presented of the medical image processing literature on mutual information-based registration. The aim of the survey is threefold: an introduction for those new to the field, an overview for those working in the field, and a reference for those searching for literature on a specific application. Methods are classified according to the different aspects of mutual information-based registration. The main division is in aspects of the methodology and of the application. The part on methodology describes choices made on facets such as preprocessing of images, gray value interpolation, optimization, adaptations to the mutual information measure, and different types of geometrical transformations. The part on applications is a reference of the literature available on different modalities, on interpatient registration and on different anatomical objects. Comparison studies including mutual information are also considered. The paper starts with a description of entropy and mutual information and it closes with a discussion on past achievements and some future challenges. PMID- 12906252 TI - Cortical surface registration for image-guided neurosurgery using laser-range scanning. AB - In this paper, a method of acquiring intraoperative data using a laser range scanner (LRS) is presented within the context of model-updated image-guided surgery. Registering textured point clouds generated by the LRS to tomographic data is explored using established point-based and surface techniques as well as a novel method that incorporates geometry and intensity information via mutual information (SurfaceMI). Phantom registration studies were performed to examine accuracy and robustness for each framework. In addition, an in vivo registration is performed to demonstrate feasibility of the data acquisition system in the operating room. Results indicate that SurfaceMI performed better in many cases than point-based (PBR) and iterative closest point (ICP) methods for registration of textured point clouds. Mean target registration error (TRE) for simulated deep tissue targets in a phantom were 1.0 +/- 0.2, 2.0 +/- 0.3, and 1.2 +/- 0.3 mm for PBR, ICP, and SurfaceMI, respectively. With regard to in vivo registration, the mean TRE of vessel contour points for each framework was 1.9 +/- 1.0, 0.9 +/- 0.6, and 1.3 +/- 0.5 for PBR, ICP, and SurfaceMI, respectively. The methods discussed in this paper in conjunction with the quantitative data provide impetus for using LRS technology within the model-updated image-guided surgery framework. PMID- 12906254 TI - Automated 3-D PDM construction from segmented images using deformable models. AB - In recent years, several methods have been proposed for constructing statistical shape models to aid image analysis tasks by providing a priori knowledge. Examples include principal component analysis of manually or semiautomatically placed corresponding landmarks on the learning shapes [point distribution models (PDMs)], which is time consuming and subjective. However, automatically establishing surface correspondences continues to be a difficult problem. This paper presents a novel method for the automated construction of three-dimensional PDM from segmented images. Corresponding surface landmarks are established by adapting a triangulated learning shape to segmented volumetric images of the remaining shapes. The adaptation is based on a novel deformable model technique. We illustrate our approach using computed tomography data of the vertebra and the femur. We demonstrate that our method accurately represents and predicts shapes. PMID- 12906255 TI - Automatic construction of 3-D statistical deformation models of the brain using nonrigid registration. AB - In this paper, we show how the concept of statistical deformation models (SDMs) can be used for the construction of average models of the anatomy and their variability. SDMs are built by performing a statistical analysis of the deformations required to map anatomical features in one subject into the corresponding features in another subject. The concept of SDMs is similar to statistical shape models (SSMs) which capture statistical information about shapes across a population, but offers several advantages over SSMs. First, SDMs can be constructed directly from images such as three-dimensional (3-D) magnetic resonance (MR) or computer tomography volumes without the need for segmentation which is usually a prerequisite for the construction of SSMs. Instead, a nonrigid registration algorithm based on free-form deformations and normalized mutual information is used to compute the deformations required to establish dense correspondences between the reference subject and the subjects in the population class under investigation. Second, SDMs allow the construction of an atlas of the average anatomy as well as its variability across a population of subjects. Finally, SDMs take the 3-D nature of the underlying anatomy into account by analysing dense 3-D deformation fields rather than only information about the surface shape of anatomical structures. We show results for the construction of anatomical models of the brain from the MR images of 25 different subjects. The correspondences obtained by the nonrigid registration are evaluated using anatomical landmark locations and show an average error of 1.40 mm at these anatomical landmark positions. We also demonstrate that SDMs can be constructed so as to minimize the bias toward the chosen reference subject. PMID- 12906256 TI - Fast algorithms for GS-model-based image reconstruction in data-sharing Fourier imaging. AB - Many imaging experiments involve acquiring a time series of images. To improve imaging speed, several "data-sharing" methods have been proposed, which collect one (or a few) high-resolution reference(s) and a sequence of reduced data sets. In image reconstruction, two methods, known as "Keyhole" and reduced-encoding imaging by generalized-series reconstruction (RIGR), have been used. Keyhole fills in the unmeasured high-frequency data simply with those from the reference data set(s), whereas RIGR recovers the unmeasured data using a generalized series (GS) model, of which the basis functions are constructed based on the reference image(s). This correspondence presents a fast algorithm (and two extensions) for GS-based image reconstruction. The proposed algorithms have the same computational complexity as the Keyhole algorithm, but are more capable of capturing high-resolution dynamic signal changes. PMID- 12906257 TI - Role of peroxisome proliferator-activated receptor gamma and its ligands in the control of immune responses. AB - To ensure that efficient immune responses against dangerous antigens are raised while tolerance to self molecules is preserved, the immune system tightly regulates activation and survival of its cellular compartments through mechanisms only partially characterized. In this context, recent evidence indicates a role in immunity of the nuclear receptor PPAR-gamma, which is upregulated in activated lymphocytes and in dendritic cells. Preliminary in vitro studies indicate that PPAR-gamma activation profoundly alters the immune properties of these cells, usually leading to the inhibition of immune responses. Naturally occurring PPAR gamma ligands include the cyclopentenone prostaglandins of the J series, which are present in bone marrow, thymus, and secondary lymphatic tissues. The levels of these metabolites are increased in inflamed tissues, where they exert strong anti-inflammatory effects leading to resolution of inflammation and wound healing. Cyclopentenone prostaglandins activate both PPAR-gamma-dependent and PPAR-gamma-independent pathways, possess intrinsic proapoptotic potential and are direct inhibitors of NF-kappaB signaling. The relevance of these effects in vivo still awaits proper evaluation in humans. Some of the newly described regulatory pathways might eventually be exploited in the treatment of immune diseases by means of PPAR-gamma ligands, such as thiazolidinediones or prostaglandins. PMID- 12906258 TI - Activation of mammalian Toll-like receptors by endogenous agonists. AB - Toll-like receptors activate innate and adaptive immune systems in mammals. This ancient family of receptors has been evolving since before the taxonomic split between the plant and animal kingdoms. The discovery of the mammalian Toll-like receptors was heralded as confirmation of a predicted biological system explicitly designed to detect exogenous molecules from micro-organisms. However, there is accumulating evidence that Toll-like receptors also detect endogenous agonists, such as the degradation products of macromolecules, products of proteolytic cascades, intracellular components of ruptured cells, and products of genes that are activated by inflammation. Here we review endogenous models of Toll-like receptor activation, a subject of extensive debate. Endogenous activation of mammalian Toll-like receptors may provide key insights for the treatment of multiple conditions, from atherosclerosis to transplant rejection. PMID- 12906259 TI - CD8 T cells and aging. AB - Aging of the immune system, or "immunosenescence," is associated with both a marked reduction in responsiveness as well as functional dysregulation. These changes have been implicated in the increased morbidity and mortality of the elderly population from infectious disease, and may also play a role in autoimmunity and cancer. Though marginal alterations in B lymphocytes are apparent, the dramatic decline in humoral and cell-mediated responses is predominantly the consequence of alterations in the T-cell compartment. The effect of aging on CD4 cell function has been extensively summarized elsewhere. This review, therefore, focuses on the CD8 T-cell subset. Age-related changes in thymic function and involution, cellular homeostasis and lifespan, population shifts, T-cell activation, the process of replicative senescence, and oligoclonal expansions are discussed in terms of their effect on CD8 T cells. Age-associated alterations in CD4 T cells and antigen-presenting cells are mentioned insofar as these cells affect CD8 T-cell activation and function. Distinct patterns of immunosenescence in humans and mice are also noted. PMID- 12906260 TI - Structure and function of the immune semaphorin CD100/SEMA4D. AB - Semaphorins are a large family of membrane-bound and secreted molecules involved in numerous functions, including axon guidance, morphogenesis, carcinogenesis, and immunomodulation. A growing number of semaphorins--namely, human CD100/SEMA4D, CD108/SEMA7A, and SEMA3A; viral semaphorins, SemaVA and SemaVB; and, very recently, mouse Sema4A--were reported to regulate immune cell responses. Among them, the role of CD100 has been well documented in both humans and mice. CD100, in particular, has been shown to influence monocyte migration, T cell activation, B-cell survival as well as T/B and T/dendritic cell cooperation. In contrast to other semaphorins, CD100 is the only semaphorin for which membrane and soluble forms are endowed with functional properties, and for which bidirectional signaling has been suggested. The human membrane-bound CD100 engagement triggers costimulatory signals to T cells through its interaction with membrane protein tyrosine phosphatase CD45 and an intracellular serine kinase. Its soluble extracellular region acts most likely through its receptors, human PlexinB1 and mouse CD72, to promote T-cell priming, B-cell survival and antibody production in response to T-dependent antigens. Human soluble CD100 also induces monocyte paralysis and the arrest of its spontaneous and chemokine-induced migration by signaling through an as yet unknown receptor that is different from PlexinB1 and CD72. In this review, we discuss recent advances in research studies on human and murine CD100, and we describe the relationship of CD100 function to its expression and structure. The signaling events that support CD100 function are also discussed. PMID- 12906261 TI - CD40 and dendritic cell function. AB - CD40 has emerged as a key signaling pathway for the function of B cells, monocytes, and dendritic cells (DC) in the immune system, and plays a major role in inflammatory pathways of nonhemopoietic cells. CD40 is expressed by monocytes and DC and is up-regulated when DC migrate from the periphery to draining lymph nodes (DLN) in response to microbial challenge. CD154 signaling by MHC restricted, activated CD4+ T cells induces differentiation of DC, as defined by an increased surface expression of MHC, costimulatory, and adhesion molecules. Thus, CD40 functions in the adaptive immune response as a trigger for the expression of costimulatory molecules for efficient T-cell activation. CD40 ligation of DC also has the capacity to induce high levels of the cytokine IL-12, which polarizes CD4+ T cells toward a T helper 1 (Th1) type, enhances proliferation of CD8+ T cells, and activates NK cells. CD40 may also play an important role in the decision between tolerance and immunity and the generation of regulatory CD4+ T cells that are thought to maintain peripheral self-tolerance in vivo. PMID- 12906263 TI - Maintenance and attrition of T-cell memory. AB - After antigenic stimulation in the context of "danger signals," naive T cells embark on a programmed, intense expansion phase to counteract the rapid proliferation of pathogens. During the first week of infection, responding T cells undergo > 1000-fold expansion, resulting in the development of large numbers of cells exhibiting potent effector function. As the pathogen (antigen) burden dwindles, a majority of the effectors generated are eliminated by apoptosis, resulting in the survival and maintenance of a small population of antigen-specific cells as long-term memory T cells. Depending on the infection studied, CD8+ T cells appear to differentiate through multiple pathways into resting and effector memory subsets, and require multiple cytokines and cell surface molecules for survival and proliferation. Once generated, the repertoire of memory T cells remains highly vulnerable to attrition during heterologous infections, where homeostatic forces drive deletions in T-cell memory pools to accommodate the entry of new memory T cells. This review will primarily focus on the factors that influence the generation, maintenance, and attrition of memory CD8+ T cells. PMID- 12906264 TI - The competition between PAOs (phosphorus accumulating organisms) and GAOs (glycogen accumulating organisms) in EBPR (enhanced biological phosphorus removal) systems at different temperatures and the effects on system performance. AB - It is well known and firmly established that the rate of chemical and biochemical reactions slow down as temperature decreases. Nevertheless, several studies have reported that the efficiency of enhanced biological phosphorus removal (EBPR) improves as temperature decreases. However, several recent studies have reported that EBPR reaction rates decrease with temperature decrease in accordance with the Arrhenius relationship. This study was designed to more thoroughly investigate this controversy using two UCT plants fed with a synthetic wastewater consisting primarily of acetate as the COD form, and a small amount of supplemental yeast extract. Experiments were performed over temperatures ranging from 5 to 20 degrees C. The results showed that, even though the kinetic rates decrease as temperature decreases, EBPR systems perform better at colder temperatures. The reason for better system performance is apparently related to reduced competition for substrate in the non-oxic zones, which results in an increased population of PAOs and, thus, greater EBPR efficiency. The proliferation of PAOs apparently occurs because they are psychrophilic whereas their competitors are not. The experiments showed that the EBPR sludges accumulated high concentrations of both PHA and glycogen at 20 degrees C, but accumulated more PHA and much less glycogen at 5 degrees C. Although the results could be interpreted as the result of changes in the PAO-GAO competition, Mann Whitney non-parametric comparisons of transmission electron microscopy examinations revealed no indication of the presence of GAOs population under any temperature conditions. Regardless, mass balances of the glycogen data showed that the involvement of glycogen is less at cold temperature, even though EBPR was greater. Unlike current EBPR models (e.g. Mino model), the results suggest that glycogen metabolism is not a precursor for EBPR biochemistry. The results also indicate that temperature not only may cause selective pressure on the dominant organisms, but also may force them to use a different metabolic pathway as temperature decreases. PMID- 12906262 TI - Signal transduction via MHC class I molecules in endothelial and smooth muscle cells. AB - MHC class I molecules have long been recognized for their ability to stimulate intracellular signals in T and B lymphocytes. More recently, it has become clear that MHC class I molecules can also initiate intracellular signals in endothelial and smooth muscle cells, which synergize with growth factor receptors to elicit cell proliferation. This review describes our current knowledge of class I mediated signaling pathways in human endothelial and smooth muscle cells. The role of the class I signaling pathway in modulating cell growth and the clinical significance of this pathway in chronic allograft rejection are discussed. PMID- 12906265 TI - Evaluation of influent prefermentation as a unit process upon biological nutrient removal. AB - The objective of this NSF sponsored research was to provide a controlled comparison of identical continuous flow biological nutrient removal (BNR) processes both with and without prefermentation in order to provide a stronger, more quantitative, technical basis for design engineers to determine the potential benefits of prefermentation to EBPR in treating domestic wastewater. Specifically, this paper focused upon the potential impacts of primary influent prefermentation upon BNR processes treating septic domestic wastewater. This study can be divided into two distinct phases--an initial bench-scale phase which treated septic P-limited (TCOD:TP>40) wastewater and a subsequent pilot-scale phase which treated septic COD-limited (TCOD:TP<40) wastewater. The following conclusions can be drawn from the results obtained to date. Prefermentation increased both RBCOD, SBCOD and VFA content of septic domestic wastewater. Prefermentation resulted in increased biological P removal for a highly septic, non-P limited (TCOD:TP<40:1) wastewater. However, in septic, P-limited (TCOD:TP>40:1) wastewater, changes in net P removal due to prefermentation were suppressed by limited P availability, even though P release and PHA content were affected. Prefermentation increased specific anoxic denitrification rates for both COD and P-limited wastewaters, and in the pilot (COD-limited) study also coincided with greater system N removal. PMID- 12906266 TI - Anoxic phosphorus removal by denitrifying heterotrophic bacteria. AB - The unexplained occurrence of anoxic phosphorus (P) accumulation has largely hampered modeling of nitrification denitrification biological excess P removal (NDBEPR) systems. The aim of this study was, therefore, to isolate and identify denitrifying-P accumulating heterotrophic bacteria (DPBs) from a NDBEPR system in order to evaluate anoxic P accumulation and the specific mechanisms involved. Results of the study showed various heterotrophic bacteria to be capable of anoxic P accumulation utilising nitrate (NO3) as electron acceptor. While Pseudomonas spp. predominated, Serratia spp. and Vibrio spp. demonstrated the most efficient anoxic P accumulation with 7.10 and 7.29 mgPO4-P/L removal, respectively, at an initial NO3 concentration of 13.54 mgNO3-N/L and P concentration of 16.34 mgPO4-P/L. Weaker DPBs were also identified which were only capable of accumulating small amounts of P at low initial P and NO3 concentrations due to weak denitrification capacity. Anoxic P release was also observed due to the presence of acetate. PMID- 12906267 TI - Quantitative estimation of the role of denitrifying phosphate accumulating organisms in nutrient removal. AB - It has been reported that a proportion of polyphosphate-accumulating organisms (PAOs) can denitrify or utilize nitrate as an electron acceptor. The usage of denitrifying-PAO (DN-PAO) can relieve the competition for COD between denitrification and phosphorus removal because they can treat nitrate and phosphate by using the same carbon source. To effectively use DN-PAO for biological nutrient removal (BNR), a new system was proposed in which an anaerobic phase is placed at the influent end, followed by he anoxic and external nitrification phase. In this study, the lab-scale proposed system (A2N system) was operated with a municipal wastewater 1) to confirm stable settlement of DN PAO in the proposed system, 2) to quantitatively estimate the fraction of different groups of organisms like denitrifiers without polyphosphate accumulating capability, aerobic-PAO and DN-PAO and 3) to estimate the advantages of DN-PAO's presence in the system. Moreover, batch experiments in which anoxic and aerobic phosphate uptake rates (PUR) were measured were also carried out. The activity of DN-PAO was observed throughout the experimental period by the batch experiment. From the results of the calculation of COD utilized by each group of organisms, it was concluded that the proposed system could accumulate much more PAO (as DN-PAO) than conventional BNR systems. Moreover, they were responsible for both EBPR and denitrification. PMID- 12906268 TI - The ScanDeNi process could turn an existing under-performing activated sludge plant into an asset. AB - With tightening up of effluent discharge standards from wastewater treatment facilities, many plants are facing costly augmentations and in many cases completely new plants will have to be constructed. The ScanDeNi process was developed in Sweden for increased nitrogen removal at the Vasteras Sewage Treatment Plant (STP), 125,000 p.e. near Stockholm, and can be described as a modified contact stabilisation process with pre-denitrification and a selector stage for nitrification. The STP was upgraded at a cost of some 25 Mill. SEK (2.5 Mill. USD). It has been successfully in operation since 1998, exceeding all expectations. The process is showing the following major advantages. 25-35% less volume for the same Sludge Retention Time (SRT) and secondary sedimentation sludge load, compared to conventional pre-denitrification; or a 25-35% higher load can be applied within the same volume with the same removal efficiencies. The selector mechanism appears to be not limited to the nitrifying bacteria alone. Other microorganisms appear to be responsible for the reduction of surface active matter from the return activated sludge (RAS), as well as in the reject stream from sludge dewatering, resulting in an increase in alpha-values of approximately 50%. Due to the high alpha-values less aeration is required, resulting in significant operating cost savings. 'Automatic' creation of anaerobic conditions, enabling biological phosphorus removal. Whilst rarely a concern in warmer climates, BNR plants in cold climates in winter often lose their capacity to nitrify. The Vasteras STP has consistently maintained excellent effluent quality even with effluent temperatures as low as 7 degrees C, and at an SRT of some 7-9 days, proving the effectiveness of the nitrifier selector. The ScanDeNi process could offer excellent effluent discharge standards (T-N < 10 mg/L, T-P < 0.5 mg/L) in smaller tank volumes and at a significantly lower operating cost, compared to conventional pre-denitrification systems. PMID- 12906269 TI - The effect of GAOs (glycogen accumulating organisms) on anaerobic carbon requirements in full-scale Australian EBPR (enhanced biological phosphorus removal) plants. AB - Glycogen-accumulating organisms (GAOs) were present in six full-scale plants investigated and in all but one made a significant contribution to the amount of volatile fatty acid (VFA) taken up anaerobically. While most plants surveyed contain GAOs, it was demonstrated that it is possible for a full-scale plant to operate with an insignificant GAO population. "Candidatus Accumulibacter phosphatis" were the significant polyphosphate-accumulating organisms (PAOs) in all plants surveyed. "Candidatus Competibacter phosphatis" were found in all plants along with other possible GAOs that were observed but not identified. A significant GAO population will increase the carbon requirements by removing VFA that could otherwise have been used by PAOs. Process optimization minimizing GAOs in full-scale plants would lead to a more efficient use of VFA. PMID- 12906270 TI - Nitrification of high strength ammonia wastewater and nitrite accumulation characteristics. AB - Biological nitrogen removal via the nitrite pathway in wastewater treatment is very important in saving the cost of aeration and as an electron donor for denitrification. Wastewater nitrification and nitrite accumulation were carried out in a biofilm airlift reactor with autotrophic nitrifying biofilm. The biofilm reactor showed almost complete nitrification and most of the oxidized ammonium was present as nitrite at the ammonium load of 1.5 to 3.5 kg N/m3 x d. Nitrite accumulation was stably achieved by the selective inhibition of nitrite oxidizers with free ammonia and dissolved oxygen limitation. Stable 100% conversion to nitrite could also be achieved even under the absence of free ammonia inhibition on nitrite oxidizers. Batch ammonium oxidation and nitrite oxidation with nitrite accumulating nitrifying biofilm showed that nitrite oxidation was completely inhibited when free ammonia is higher than 0.2 mg N/L. However, nitrite oxidation activity was recovered as soon as the free ammonia concentration was below the threshold level when dissolved oxygen concentration was not the limiting factor. Fluorescence in situ hybridization analysis of cryosectioned nitrite accumulating nitrifying biofilm showed that the beta-subclass of Proteobacteria, where ammonia oxidizers belong, was distributed outside the biofilm whereas the alpha-subclass of Proteobacteria, where nitrite oxidizers belong, was found mainly in the inner part of the biofilm. It is likely that dissolved oxygen deficiency or limitation in the inner part of the nitrifying biofilm, where nitrite oxidizers exist, is responsible for the complete shut down of the nitrite oxidizers activity under the absence of free ammonia inhibition. PMID- 12906271 TI - Enhanced nitrogen removal in SBRs bypassing nitrate generation accomplished by multiple aerobic/anoxic phase pairs. AB - A lab-scale SBR was used for the study of nitrogen removal from a synthetic wastewater with an ammonium-nitrogen concentration of 50 mg/L. The react phase of the reactor operation was divided into three sets of consecutive aerobic and anoxic periods with a duration ratio of 1:3 (20 min aerobic and 1 h anoxic phase). Under these operating conditions, nitrogen removal was achieved via nitrite i.e. no nitratification (oxidation of nitrite to nitrate) and hence no denitratification (reduction of nitrate to nitrite) was taking place in the aerobic and anoxic phase, respectively. This was attributed to the suppression of the nitrite-oxidizers activity due to the short aerobic phase duration. This presumption was supported by the ever decreasing amount of nitrate-nitrogen generated in the react phase during the transient, even when the activated sludge of the reactor was supplemented with additional nitrite-oxidizers. On the other hand, denitrification was mainly based on stored carbon sources, as long as the organic carbon (provided in the form of acetate) was never accumulated during the anoxic/anaerobic fill phase of the reactor operation. PMID- 12906273 TI - Fuzzy-control for improved nitrogen removal and energy saving in WWT-plants with pre-denitrification. AB - In the last few years, numerous studies were carried out, dealing with the application of fuzzy-logic to improve the control of the activated sludge process. In this paper, fuzzy-logic based control strategies for wastewater treatment plants with pre-denitrification are presented that should lead to better effluent quality and, in parallel, to a reduction of energy consumption. Extensive experimental investigations on a large scale pilot plant as well as simulation studies (ASM1 with SIMBA) were carried out in order to design, evaluate and compare different fuzzy-controllers with each other and with comparable conventional control systems. The fuzzy-controllers were designed as high-level controllers that determine the DO-setpoints in the aerated zones and the ratio between aerated and non-aerated zones. Conventional PI-controllers were used to maintain the DO-concentration at the set-point levels. The ammonia and nitrate concentration in the effluent and the ammonia load in the influent were considered as input variables for the different fuzzy-controllers. Compared to the operation with fixed nitrification/denitrification zones and constant DO concentrations, the required air-flow could be reduced up to 24% by using fuzzy logic based control strategies. In comparison with a more advanced conventional control strategy (relay controller with two thresholds and the NH4-N concentration in the effluent as single control variable) a reduction of air-flow rate up to 14% could be achieved. At the same time, NH4-N peaks in the effluent that are normally caused by peak flow conditions could be reduced significantly. The large scale experiments show that the fuzzy-controllers can be easily implemented in modern control and supervision systems and that the control characteristics can be followed and modified during operation. It therefore can be expected that the developed fuzzy-control systems will be accepted by the operating personnel in wastewater treatment plants. PMID- 12906272 TI - Induction of denitrification in a pilot-scale trickling filter by adding nitrate at high loading rate. AB - Oxygen transferred from the liquid phase into a biofilm can be used for aerobic degradation of organic matter and for nitrification. A second possible pathway for the removal of organic matter is denitrification in anoxic zones deeper in the biofilm. At high organic loading rates with insufficient oxygen supply to the biofilm, denitrification could be induced by providing the biofilm with external nitrate. This possibility was tested in a pilot-scale trickling filter by adding a pulse of nitrate to a highly loaded trickling filter. The experiment showed that denitrification can indeed be induced by adding nitrate at high loading conditions and that this way a considerably increased substrate removal capacity can be obtained. The fact that denitrification occurred was confirmed by the increased production of CO2 from bioconversion processes, without a major change of the O2 consumption. The simplified mixed-culture biofilm model developed by Rauch et al. was extended for the description of off-gas measurements and was able to describe the results of the experiment very well. PMID- 12906274 TI - Use of pH as fuzzy control parameter for nitrification under different alkalinity in SBR process. AB - In order to achieve fuzzy control of nitrification in a Sequencing Batch Reactor (SBR) brewery wastewater was used as the substrate. The effect of alkalinity on pH variation during nitrification was systematically studied, at the same time the variations of DO and ORP were investigated. Alkalinity and pH of the wastewater were adjusted by adding sodium bicarbonate at five levels and sodium hydroxide at two levels. Unadjusted wastewater was also studied. According to the results, variation of pH could be divided into rising type and descending type. When bicarbonate alkalinity was deficient or sufficient, the descending type happened. If alkalinity was deficient, the pH decreasing rate got slower when nitrification nearly stopped; if alkalinity was sufficient, at the end of nitrification pH turned from decrease to increase. This was the most common situation and pH could be used to control the end of nitrification. When alkalinity was excessive, the rising type happened, pH was increasing at nearly a constant rate during and after nitrification and could not be used to control the nitrification time, but if the aeration rate was moderate DO could be used to control the nitrification time. This situation seldom happened. Therefore the variation of pH could not only be used to control the nitrification time but also to judge whether the alkalinity was enough or not. On the basis of this, the fuzzy controller of nitrification in SBR was constructed. When discussing the influence of pH on nitrification rate the composition and concentration of alkalinity must be considered or else the results may be incomprehensive. And to some extent the influence of alkalinity on nitrification rate was more important than pH. PMID- 12906275 TI - Nitrification preservation in activated sludge during curative bulking chlorination. AB - The bulking that occurs in biological wastewater treatment plants using activated sludge is very often controlled by the injection of sodium hypochlorite into the return activated sludge (RAS) stream. In the present study undertaken at two pilot plants fed with synthetic wastewater, the impact of the pass frequency of the sludge at the chlorine dosing point on the nitrifying flora is analysed. The pass frequency is one for the pilot plant 1 and two for the pilot plant 2. A dose of chlorine of 4.85 +/- 0.05 g/kg/MLVSS per day was applied at both pilots. The preservative effect on nitrifying activity of the lowest concentration of chlorine at the dosing point and therefore of the highest pass frequency was evidenced. Among other tools, a simple method of measurement of the oxygen uptake rate enabled us to monitor the effect of chlorination on nitrification before recording an increase in the ammonia concentration in the bulking. PMID- 12906276 TI - Partial nitrification in a high-load activated sludge system by biofilter backwash water recirculation. AB - The South-Budapest Municipal Wastewater Treatment Plant (SBWWTP) based on the high-load activated sludge process (AS) was upgraded into nutrient removal in 1998-1999 in Hungary. Biofor type nitrifying (NP) and denitrifying (DN) biofilters were implemented for nitrogen removal downstream of the (AS) stage in order to meet the required effluent standards characterized by 50 mgCOD l(-1), 10 mgBOI5 l(-1), 10 mgTN l(-1), 1 mgTP l(-1) and 35 mgTSS l(-1). Phosphorus removal is obtained by chemical addition. The study presented sums up the performance of the upgraded plant throughout 25 months. Besides the efficient pre denitrification obtained in the AS basin, significant ammonium oxidation was observed in the aerated zone, upon the introduction of recirculation of nitrate rich water and backwash water. The introduction of biofilter backwash water, containing nitrifying biomass, has generated significant ammonium conversion in the aerated basins, where nitrification was observed previously to a very limited extent. Process parameters of the AS stage were: aerobic sludge age of 1.5-2.0 days, 1.5 hours hydraulic retention time, specific sludge load of 1.1 kgCOD kgMLSS(-1) d(-1). The nitrification in the AS tank has significantly reduced below 16 degrees C, however, denitrification efficiency was not as highly dependent upon temperature during the investigated period. The paper investigates the nitrogen balance of the plant in two aspects: (i) the effects of seeding on nitrification in the high load AS reactor and (ii) pre-denitrification potential in the anoxic zone of the AS tank of the pre-settled wastewater. PMID- 12906277 TI - Nutrient removal from wastewaters using high performance materials. AB - Return side streams from anaerobic digesters and dewatering facilities at wastewater treatment plants (WWTPs) contribute a significant proportion of the total nitrogen load on a mainstream process. Similarly, significant phosphate loads are also recirculated in biological nutrient removal (BNR) wastewater treatment plants. Ion exchange using a new material, known by the name MesoLite, shows strong potential for the removal of ammonia from these side streams and an opportunity to concurrently reduce phosphate levels. A pilot plant was designed and operated for several months on an ammonia rich centrate from a dewatering centrifuge at the Oxley Creek WWTP, Brisbane, Australia. The system operated with a detention time in the order of one hour and was operated for between 12 and 24 hours prior to regeneration with a sodium rich solution. The same pilot plant was used to demonstrate removal of phosphate from an abattoir wastewater stream at similar flow rates. Using MesoLite materials, >90% reduction of ammonia was achieved in the centrate side stream. A full-scale process would reduce the total nitrogen load at the Oxley Creek WWTP by at least 18%. This reduction in nitrogen load consequently improves the TKN/COD ratio of the influent and enhances the nitrogen removal performance of the biological nutrient removal process. PMID- 12906278 TI - Controlling nitrogen removal using redox and ammonium sensors. AB - At the Ejby Molle WWTP (300,000 PE) operated by the Odense Water Ltd. a system of aeration control which combines ammonium concentration and redox potential has proven itself as a method of reducing nitrogen levels in the effluent. The nitrogen removal process at this plant proceeds in parallel aeration tanks that alternate between periods of aeration and denitrification. At the same time influent is redirected from one tank to the other such that influent and effluent is primarily to and from the tank where denitrification is going on. The aeration stops and the tanks open and close when the ammonium concentration reaches its set point. Aeration is restarted again when redox potential drops to its low set point or when ammonium reaches its upper set point. The average total nitrogen concentration in the effluent from the Ejby Molle WWTP went from 4.7 down to 2.1 mgN/l after this control system was implemented. Not all of this reduction can be explained by better control. However we believe that at least 1 mgN/L of the reduction and probably more was the result of the control system based on the combination of ammonium and redox sensors. PMID- 12906279 TI - Design of nutrient removal activated sludge systems. AB - A mechanistic mathematical model for nutrient and organic matter removal was used to describe the behavior of a nitrification denitrification enhanced biological phosphorus removal (NDEBPR) system. This model was implemented in a user-friendly software DESASS (design and simulation of activated sludge systems). A 484-L pilot plant was operated to verify the model results. The pilot plant was operated for three years over three different sludge ages. The validity of the model was confirmed with data from the pilot plant. Also, the utility of DESASS as a valuable tool for designing NDEBPR systems was confirmed. PMID- 12906280 TI - A thermal adaptation of bacteria to cold temperatures in an enhanced biological phosphorus removal system. AB - Temperature is one of the key parameters that affects the reaction kinetics and performance of enhanced biological phosphorus removal (EBPR) systems. Although studies agree regarding the effect of temperature on kinetic reaction rates, there are contradictory results in the literature regarding the effect of temperature on EBPR system performance. Early investigators (Sell, Ekama et al., Daigger et al.) reported better performance with lower temperatures, but others have reported partial or complete loss of EBPR functions at low temperatures (McClintock et al., Brdjanovic et a., Beatons et al.). One speculation is that deterioration in the EBPR system performance at cold temperatures can be attributed to rigid-like behavior of the cell membranes. Most cells (not all) on the other hand have the ability to alter their membrane fatty acid composition as temperature changes in order to keep their membrane at nearly the same fluidity despite the temperature changes. This unique ability is known as homeoviscous adaptation. In this study, homeoviscous adaptation by EBPR activated sludge was investigated for a series of temperatures ranging from 20 degrees C to 5 degrees C using a lab scale continuous flow EBPR system fed with acetate and supplemental yeast extract. The fatty acid analysis results suggested that the unsaturated to saturated fatty acid ratio increased from 1.40 to 3.61 as temperature dropped from 20 to 5 degrees C. The increased cis-9-hexadecanoic acid (C 16:1) at 5 degrees C strongly indicated the presence of homeoviscous adaptation in the EBPR bacterial community. Thus the cell membranes of the EBPR community were still in a fluid state, and solute transport and proton motive force were operable even at 5 degrees C. It was concluded that loss of EBPR performance at low temperatures is not related to the physical state of the cellular membranes, but is possibly related to the application of unsuitable operational conditions (low SRT, excessive electron acceptors, low anaerobic detention time, non-acclimated sludge, etc.). PMID- 12906281 TI - Nitrification in biofilters under variable load and low temperature. AB - The evolution of European legislation has led to the rehabilitation of many wastewater treatment plants, sometimes through the installation of a biological complementary treatment stage. Among these sites, some plants in mountain areas are considering a biofiltration process. The design of such plants, especially for winter, appears to be tricky because of the very low influent temperature, the high performance requested for ammonia removal and the important and short term variations of the influent loads. The monitoring of a site during two consecutive winters has allowed us to study some aspects of the treatment. The major results are: a maximal nitrification capacity of about 0.59 kg of formed N NO3- x m(-3) of material x d(-1) with an influent temperature around 7 degrees C at the plant inlet; a nitrifying biomass growth rate, expressed as nitrifying capacity increase, of 0.03 kg of N-NO3- x m(-3) of material x d(-2); quick and short terms load variations require a specific operation of the filters prior to the load increase, in order to grow enough active biomass to be able to treat the peak load immediately. PMID- 12906283 TI - Fenton's reagent and coagulation-flocculation as pretreatments of combined wastewater for reuse. AB - In Mexico City, drinking water is mainly produced from groundwater (70%). This practice has caused collateral problems such as Mexico City's soil sinking (5-30 cm/year). One of the most viable alternatives to palliate this problem is the treatment of wastewater for reuse in either irrigation or for groundwater artificial recharge. This paper presents the evaluation of two physicochemical pretreatment systems to treat the wastewater from the metropolitan area of the Mexican Valley that are conducted by two main sewage systems called Great Canal and Churubusco River. In this research two treatment processes were studied: 1) coagulation-flocculation and, 2) Fenton's reagent. For each one of these processes suggested, tests were performed with wastewater samples of the Great Canal and the Churubusco River mixed in a volume ratio of 1:1. In the case of the coagulation-flocculation process, additional experiments were performed to determine the optimal conditions by applying an experimental design technique. In this experimental design, six coagulant agents were considered (alum, ferric chloride, three coagulant reagents of polymeric kind with aluminium and a coagulant reagent of natural origin), and three flocculant agents (an anionic, a cationic, and a non ionic polymers). Concerning the application of the Fenton's reagent (Fe2+:H2O2), the experimental variables were the weight ratio of the ferrous iron and the hydrogen peroxide and the concentrations of these reagents. The pH value was controlled to be near to 4. For the best experimental conditions, the effluent of the Fenton's method showed similar physicochemical characteristics to the wastewater treated by coagulation-flocculation. Nevertheless, Fenton's reagent showed two very important advantages compared to the coagulation-flocculation process: a disinfecting effect and a lower production of residual sludges. PMID- 12906284 TI - Nitrification and denitrification using a single biofilter packed with granular sulfur. AB - This study was performed to develop a granular sulfur packed nitrification/denitrification process employing a uniquely designed single biofilter, which treated a relatively low carbon loaded domestic wastewater taken from a primary clarifier at a municipal wastewater treatment facility. The system was tested on varying experimental conditions, e.g. inflow flow, organic load and nitrogen load. Regardless of flow rate being increased, SS and COD was unvaryingly removed up to 90 and 80%, respectively. Moreover, TKN was also decomposed up to 90%. Increase in COD load gradually led to escalating level of non-biodegradable compounds observed in effluent. Nitrification was accomplished as high as 92%, whereas denitrification was achieved up to approximately 87%. For a while, nitrification and denitrification were observed at 0.65 and 0.55 kg/m3 x day, respectively. Eventually, T-N was decomposed as high as 46%. It was concluded that granular sulfur can be used for not only electron donor, but also for a media to properly treat low carbon loaded wastewater and to filter SS efficiently. PMID- 12906282 TI - Adaptive control of the nitrate level in an activated sludge process. AB - In an activated sludge process for nitrogen removal, nitrate may be reduced to nitrogen gas by facultative heterotrophic bacteria in an anoxic environment. In order to guarantee sufficient supplies of readily biodegradable carbon compounds, an external carbon source often needs to be added. In this paper, an automatic control strategy for controlling the nitrate level using an external carbon source is presented. The external carbon source is added in the first anoxic zone to control the nitrate concentration in the last anoxic zone. Key process parameters are estimated on-line for a simplified Activated Sludge Model No 1. The estimated parameters are used for updating a linear quadratic controller. The strategy is illustrated in a simulation study with realistic influent data and is shown to perform very well. PMID- 12906286 TI - Molecular biological methods (DGGE) as a tool to investigate nitrification inhibition in wastewater treatment. AB - Incomplete nitrification at an activated sludge plant for biological pre treatment of rendering plant effluents led to a detailed investigation on the origin and solution of this problem. Preliminary studies revealed that an inhibition of ammonia oxidising microorganisms (AOM) by process waters of the rendering plant was responsible for the situation. We were able to show a correlation between the existence of specific AOM and nitrification capacity expressed as oxygen uptake rate for maximal nitrification (OURNmax). Only Nitrosospira sp. was found in the activated sludge of the rendering plant and another industrial wastewater treatment plant with problems in nitrification, while reference plants without nitrification problems showed Nitrosomonas spp. as the predominant ammonia oxidising bacteria. By accompanying engineering investigations and experiments (cross-feeding experiments, operation of a two stage laboratory plant) with molecular biological methods (DGGE--Denaturing Gradient Gel Electrophoresis) we were able to elaborate an applicable solution for the rendering plant. Laboratory experiments with a two-stage process layout finally provided complete nitrification overcoming the inhibiting nature of process waters from the rendering plant. DGGE analysis of the second stage activated sludge from the laboratory plant showed a shift in population structure from Nitrosospira sp. towards Nitrosomonas spp. simultaneous to the increase of nitrification capacity. Nitrification capacities comparable to full-scale municipal wastewater treatment plants could be maintained for more than two months. As the design of wastewater treatment plants for nitrification is linked to the growth characteristics of Nitrosomonas spp., established criteria can be applied for the redesign of the full-scale plant. PMID- 12906285 TI - Phased upgrading for nitrogen removal--a low cost approach. AB - A phased approach has been adopted for upgrading the Luggage Point WWTP for nitrogen removal. Luggage Point WWTP (900,000 EP) is the largest plant operated by Brisbane Water. Management actions recommended by the South East Queensland Wastewater Management Strategy require the effluent total nitrogen levels to be reduced to less than 5 mg/L (50th percentile) by 2005. Luggage Point WWTP originally featured primary settling and a completely nitrifying activated sludge process. Effluent total nitrogen levels from the plant averaged 27 mg/L. Characteristics of the wastewater are highly variable, with primary settled wastewater having a COD/BOD5 ratio averaging 2.5, a TKN/COD ratio averaging 0.12 and effluent non-ammonia TKN averaging 1.8 mg/L (but up to 5 mg/L). The upgrade presented a substantial challenge, due to the limited available substrate for denitrification, a significant level of unbiodegradeable nitrogen and the low existing bioreactor HRT. The phased upgrading strategy was adopted to overcome these challenges most effectively and at low cost. The first stage of the upgrade included modifications to the existing plant infrastructure, without constructing new tankage. The existing bioreactors have been retrofitted using a five stage Bardenpho nutrient removal process, incorporating significant flexibility. The first phase upgrade has been commissioned and preliminary results indicate that the effluent total nitrogen has been reduced below the required first phase target of 8 mg/L. This represents a reduction of 3 tonnes of nitrogen per day entering Moreton Bay. The process is also operating with good stability and with excellent sludge settleability. The first phase of the upgrade will be completed for 25M AUS dollars, which represents a low cost of 28 dollars per equivalent person. Further upgrade works are likely to include separate treatment of the digested sludge dewatering centrate and advanced control of the biological process. The upgrade of Luggage Pt WWTP has shown that low effluent nitrogen can be achieved at low cost, through optimum utilisation of existing assets and a phased approach to modifications. PMID- 12906288 TI - Nitrogen removal in industrial wastewater by nitration and denitration--3 years of experience. AB - CPKelco ApS, Denmark is the largest pectin plant in the world and the second largest refined carrageenan plant. The products are used for texturising purposes, primarily within the food industry, but also within the pharmaceutical industry. The products are extracted from imported natural raw materials, like dried citrus peel and special seaweed plants. In the production processes a considerable amount of water and energy are used. The excess water from the production processes is led to CPKelco's own WWTP, which is one of the largest industrial WWTPs in Denmark. In order to obtain higher process stability and lower energy consumption in the WWTP, CPKelco decided to change the nitrogen removal process from a conventional nitrification/denitrification process to a nitration/denitration process, which comprises an oxidation of ammonium to nitrite and a controlled reduction of nitrite to N2. Theoretically this process will decrease the oxygen consumption for oxidation by 25% and the use of carbon source for the reduction will be decreased by 40% compared to the conventional process. This paper presents and discusses the experiences and results from three year's continuous operation of the nitration/denitration process in an activated sludge plant, and the overall performance results are discussed in relation to the previous results. Accordingly the implementation of the nitration/denitration process was done successfully, and today the plant operates with much higher process stability than obtained before, and even the most stringent effluent requirements for nitrogen can be obtained. PMID- 12906289 TI - Development of a two-stage flexible fibre biofilm reactor for treatment of food processing wastewater. AB - Biofilm (or attached growth) reactors can be effectively used to treat organic wastewater from various industries such as food processing industry. They have a number of advantages including high organic loading rates (OLRs) and improved operational stability. A flexible fibre biofim reactor (FFBR) has been developed for efficient and cost effective treatment of food processing wastewater. In the process, simple flexible fibre packing with a very high specific surface area is used as support for microorganisms. The COD removal efficiencies for a range of OLRs have been studied. The FFBR can support an increasingly high OLR, but with a corresponding decrease in the COD removal efficiency. Therefore, a two-stage FFBR was developed to increase the treatment efficiency for systems with high OLRs. Experimental results indicated that a high overall COD removal efficiency could be achieved. At an influent COD of about 2700 mg/L and an OLR of 7.7 kgCOD/m3d, COD removal efficiencies of 76% and 82% were achieved in the first and the second stage of the reactor, respectively. The overall COD removal efficiency was 96%. Therefore, even for wastewater samples with high organic strength, high quality treated effluents could be readily achieved by the use of multiple stage FFBRs. PMID- 12906287 TI - Pre-nitrification by encapsulated nitrifiers--a possibility for self-sufficient energy operation of domestic WWTPs. AB - The overall energy consumption of domestic wastewater treatment plants (WWTPs) increases with treatment efficiency. Approximately 30 to 45 kWh per people equivalent and year is mostly necessary for advanced nitrogen and phosphorus removal, while the aeration contains the main part of approximately 60%. A new process using encapsulated nitrifiers on gel lens beads is introduced to overcome the high energy consumption of aeration. A more selective nitrification process was found at a nitrification rate of between 50 and 60 mg nitrogen per hour and litre reaction volume corresponding to a hydraulic retention time (HRT) of about 30 to 60 minutes while the soluble Chemical Oxygen Demand (COD) removal could be less than 30% depending on operational conditions of the bio-reactor. The latter enables internal use of wastewater's COD for a post denitrification. For the new process the energy consumption as well as total volume of bio-reactor are much less (approximately 30 to 50% for both) than conventional processes due to the low sludge age for COD and nitrate removal and the avoidance of internal wastewater recycle. Therefore, self-sufficient energy operation of domestic WWTPs operating with advanced treatment efficiency could become possible, if energy recovery by anaerobic sludge digestion is included. PMID- 12906290 TI - Bacteria reduction and nutrient removal in small wastewater treatment plants by an algal biofilm. AB - Attached algae settlement is frequently observed in effluents of wastewater treatment plants at locations with sufficient sunlight. For their growth they incorporate nutrients and the surface of the algal biofilm accumulates suspended solids from the clarified wastewater. During the photosynthesis process of algal biofilms oxygen is produced while dissolved carbon dioxide is consumed. This led to an increasing pH due to the change of the carbon dioxide equilibrium in water. The high pH causes precipitation of dissolved phosphates. Furthermore an extensive removal of faecal bacteria was observed in the presence of algae, which may be caused by the activity of algae. The experimental results indicate the high potential of these attached algae for polishing secondary effluent of wastewater treatment plants. Especially for small wastewater treatment plants a post connected stage for nutrient removal and bacteria reduction can be developed with the aid of an algal biofilm. PMID- 12906292 TI - Efficiency of the Activated Sludge Model no. 3 for German wastewater on six different WWTPs. AB - In 1999, the Activated Sludge Model No. 3 by the IWA Task Group on Mathematical Modelling for the Design and Operation of Biological Wastewater Treatment was presented. The model is used for the simulation of nitrogen removal. The simulations in this paper were done on the basis of a new calibration of the ASM 3 by Koch et al., with the easily degradable COD measured by respiration. For modelling of EBPR the BioP-Module of Rieger et al., was used. Six German wastewater treatment plants were simulated during this research to test the existing set of parameters of the models on various large scale plants. It was shown that changes for nitrification and enhanced biological phosphorus removal in the set of biological parameters were necessary. Sensible parameters and recommended values are presented in this article. Apart from the values of the changed biological parameters, we will in our examination discuss the modelling of the different activated sludge systems and the influent fractioning of the COD. Two plants with simultaneous denitrification in the recirculation ditch (EBPR) are simulated, one with preliminary dentrification, one with intermittent denitrification (EBPR), one with cascade denitrification (EBPR), and one pilot plant according to the Johannesburg-process (EBPR) which was simulated over a period of three months. PMID- 12906291 TI - The effect of low sludge age on wastewater fractionation (S(S), S(I)). AB - In lab-scale experiments at the 2-stage activated sludge pilot plant of Vienna's central WWTP it is shown that the wastewater soluble COD concentration, which is inert to a sludge with SRT < 1 d (SI(A)) is about double compared to the S(I) concentration in sludge with SRT > 10 d (SI(B)). Unexpectedly the ratio of SI(A)/SI(B) is independent of the sludge age between SRTs of 0.4 and 1.0 days. The difference between the two S(I) fractions is soluble COD that is readily biodegradable by the sludge with SRT > 10 d. However, it is degraded at a lower maximum growth rate. These results comply with earlier results gained with different methods and at different WWTPs. It is hypothesised that very low sludge ages result in a selection of fast growing bacteria, which can utilise only part of the S(S) in the raw wastewater. The other part of S(S) therefore remains in the wastewater and can thus be utilised for enhanced denitrification in the second stage. It is still unknown beyond which sludge age the soluble inert COD SI(A) starts to decrease, finally reaching the value SI(B) for low loaded systems (SRT > 5 days). From this point on S(I) and S(S) are assumed only to depend on the wastewater composition and not on the sludge age. The assumption of the Activated Sludge Model No.1 that the biodegradable fractions can be modelled as a single substrate and by a single removal kinetic (one Monod term) appears not to be applicable for low sludge ages. Some suggestions for mathematical modelling, design and operation of 2-stage activated sludge systems are given. PMID- 12906293 TI - Modification of Activated Sludge Model no. 3 considering direct growth on primary substrate. AB - This paper provides the structural framework for the proposed modified version of Activated Sludge Model No. 3 (ASM3), where direct heterotrophic growth on readily biodegradable substrate is included as a new process and provision is made so that growth on internal storage compounds is started sequentially, after the depletion of the external primary substrate pool. The results have provided strong indication that there was a need for considering direct growth on primary substrate as a significant biological mechanism. PMID- 12906294 TI - Polyhydroxyalkanoate form and polyphosphate regulation: keys to biological phosphorus and glycogen transformations? AB - Experimental studies with both synthetic and real domestic wastewater showed that poly-3-hydroxy-butyrate (3HB) and poly-3-hydroxy-valerate (3HV) formed in direct proportion to the acetate/propionate (Ace/Pro) ratio of the influent wastewater during Enhanced Biological Phosphorus Removal (EBPR). Acetic acid resulted in higher anaerobic phosphorus (P) release, polyhydroxyalkanoate (PHA) yield, 3HB content, and glycogen (CH) degradation. Linear regression showed that anaerobic P release (Prel) and CH degradation (CHdeg) were both a function of Ace-->3HB, but not of Pro-->3HV. Aerobic P uptake (Pup) correlated best with preceding Prel, rather than PHA (but note Prel correlated with Ace-->3HB). Aerobic CH formation (CHform) correlated best with CHdeg and 3HB. The results imply the acetate/propionate content of influent has a major influence on PHA, CH, and P transformations. Short-term increases in acetic or propionic acid increased Prel, but were always offset by corresponding changes in Pup to yield the same net P removal as the control reactor. Thus net P removal, and EBPR process performance, was probably a function of the population selected (i.e. XPAO fraction) during long-term cultivation. PMID- 12906295 TI - The effect of organic loading rate on the aerobic granulation: the development of shear force theory. AB - The effect of organic loading rate (OLR) on aerobic granulation was studied by adopting three column-shaped, sequential aerobic sludge blanket reactors (SASBR). The reactors had been fed with laboratory prepared, synthetic dextrose-nutrient broth substrate. Experimental results showed clearly that the formation, characteristics and stability of aerobic granules had a close relationship with the strength of OLR applied. Aerobic granules appeared firstly under the OLR of 4 kg COD x (m3 x day)(-1). The system stabilization was demonstrated by its little changed amount and morphology of granules. The characteristics of the stabilized granules were: 5.4 mm in mean diameter, 1.29 in roundness, 118 mg O2 x (mg VSS x hr)(-1) in SPOUR. The respective biomass SVI was 50 mL x (g MLVSS)(-1) and the averaged COD removal rate was 95%. Under the OLR of 8 kg COD x (m3 x day)(-1), granules appeared two days later than those for 4 kg COD x (m3 x day)(-1) and they always coexisted with flocs. The formed granule bed was not as compact as that under 4 kg COD x (m3 x day)(-1). There were no granules formed under the OLR of 1 kg COD x (m3 x day)(-1). Instead, flocs with rather loose structure dominated reactor mixed-liquor. The respective SVI's were 65 and 138 mL x (g MLVSS)(-1) under OLR of 8 and 1 kg COD x (m3 x day)(-1). It was proposed that the growth and maintenance of aerobic granules follow the shear force balance theory. Under the OLR of 4 kg COD x (m3 x day)(-1), a balance was reached between the aeration shear force and organic loading rate. Under this favored condition aerobic granules formed quickly and, became stabilized with the experimental parameters remained unchanged. PMID- 12906296 TI - Elevated precursor 16S rRNA levels suggest the presence of growth inhibitors in wastewater. AB - Conventional activated sludge systems require bacteria to grow to avoid washout through decay and routine solids wasting. Recently we developed a procedure targeting precursor 16S ribosomal RNA to measure the in situ growth activity of phylogenetically defined microbial populations, and this procedure was used to study the growth of bacteria in activated sludge systems. The current study significantly expands this previous work by quantifying levels of precursor 16S ribosomal RNA within individual cells of pure cultures of bacteria exposed to various culture conditions. Initially, three ranges (Type I, Type II, and Type III) of precursor 16S ribosomal RNA levels were defined by whole cell fluorescence in situ hybridization of a pure culture of Acinetobacter calcoaceticusT prepared in three culture conditions. Low levels of precursor 16S ribosomal RNA (Type I) corresponded to a stationary phase culture prepared overnight in Luria-Bertani medium. Intermediate levels of precursor 16S ribosomal RNA (Type II) corresponded to a culture transferred into fresh Luria-Bertani medium, and high levels of precursor 16S ribosomal RNA (Type III) corresponded to a culture treated with the growth inhibiting antibiotic chloramphenicol. Subsequently, the abundance of individual cells of each Type were measured in four different pure cultures after exposure to 0.45-microm filtered primary effluent collected from four different conventional activated sludge treatment plants in Cincinnati, OH, USA. Individual cells of each Type were observed in the culture of A. calcoaceticusT exposed to each of the four primary effluents. Only Type I cells were observed in cultures of A. johnsoniiT, A. johnsonii strain 210a, and Escherichia coliT exposed to each of the four primary effluents. These results suggest that the growth of A. calcoaceticusT was inhibited by an unidentified component of filtered primary effluent present in each of the four wastewaters; whereas the growth of A. johnsoniiT, A. johnsonii strain 210a, and E. coliT were not inhibited. These results have significance for understanding the growth of phylogenetically defined microbial populations within activated sludge treatment systems. If the pattern of elevated p16S rRNA levels observed in A. calcoaceticusT is prevalent in many microbial populations in activated sludge systems, this may have implications for preventing washout of critical microbial populations that may be experiencing growth inhibition. PMID- 12906297 TI - Modelling the effect of shear history on activated sludge flocculation. AB - The aim of this paper is to investigate the effect of shear history on activated sludge flocculation dynamics and to model the observed relationships using population balances. Activated sludge flocs are exposed to dramatic changes in the shear rate within the treatment process, as they pass through localised high and low mixing intensities within the aeration basin and are cycled through the different unit operations of the treatment process. We will show that shear history is a key factor in determining floc size, and that the floc size varies irreversibly with changes in shear rate. A population balance model of the flocculation process is also introduced and evaluated. PMID- 12906298 TI - Improving titrimetric techniques by modelling pH change in activated sludge systems. AB - Existing titrimetric techniques rely on a well defined hydrogen ion production rate. In particular, they are only suitable in circumstances in which constant background pH influencing reactions take place. This is rarely the case due to the presence of the carbonate acid/base system. In this paper, a model, which describes the influence of the nitrification process on pH and accounts for the action of the carbonate system, is presented. The validity of the model is tested by comparison of model predictions for the important state variables with that of experimental data from a batch oxidation of ammonium nitrogen. The two cases studied are the responses of an endogenously respiring nitrifying sludge to: an ammonium chloride pulse and a pulse of both bicarbonate and ammonium chloride. The results are most encouraging as the dynamic HPR response is mirrored by the model simulation. Furthermore, using the model for data interpretation, the initial nitrogen substrate levels are recovered. It is shown that this could not have been achieved in either case using existing titrimetric techniques. PMID- 12906299 TI - Structure-function dynamics and modeling analysis of the micro-environment of activated sludge floc. AB - Biodegradation by microorganisms and mass transfer resistance in the micro environment of activated sludge floc can cause changes in substrate and dissolved oxygen concentrations within the floc and can contribute to stratification of microbial processes inside the flocs. In this study, an integrated model of the microenvironment of the activated sludge floc was developed for floc from wastewaters from several sources and of varying strengths for dynamic simulation of the combined biological processes of COD and nitrogen removal. The model simulation results and measured profiles show the heterogeneous and gradient governed microenvironment of activated sludge floc under different substrate and bulk oxygen concentrations. The substrate concentration increase zones inside the floc were present in all activated sludge floc from the Miller Brewing Co. wastewater treatment facility (high pollutant strength), with an oxygen penetration depth of only 0.15 mm into the outer layer. The anoxic and substrate concentration increase zones also dominated in the activated sludge floc from the Mill Creek Plant influent (medium pollutant strength), with the outer layer (0.20 mm) participating in the metabolism of the pollutants. The radius of the substrate concentration increase zone inside the sludge floc decreased with pollutant removal along the length of the tank. When the pollutant concentration in the bulk wastewater was low (Muddy Creek Plant), the substrate concentration increase zone disappeared; the whole floc was aerobic and in a high redox status. Our experiments and model analyses demonstrate that the microorganisms' structure functions inside activated sludge floc change with the bulk substrate concentration and dissolved oxygen concentration. PMID- 12906300 TI - Dimensional analysis for establishing the testing criteria of kinetic study with respirometry. AB - The kinetic study of a microbial system requires the determination of kinetic parameters under a set of operating variables. Previous researchers indicated that initial conditions, such as S0/X0 and S0/KS may influence the reliability of the parameter estimation. However, little study has been done to identify the sensitivity of system variables to the parameter estimation. This study proposes a novel dimensional analysis and identifies six dimensionless groups: mu(m)/fw, kd/fw, Yg, S0/KS, KS/S0, and 1/(fw theta(c) - 1). By incorporating the SP-moving algorithm proposed by Wu and coworkers in 2001, an algorithm was proposed in this study to perform a sensitivity analysis on the six dimensionless groups. Results of this analysis reveal that S0/X0 is more sensitive than S0/KS, as also evidenced by the fact that gross growth yield (Yg) is sensitive and affecting S0/X0. The analysis also suggests that the theta(c)-based wasting frequency (fw theta(c)) is more sensitive than the daily wasting frequency (fw). A critical minimum value of 1.3 for S0/X0 and a maximum value of 0.1 for S0/KS were suggested to establish the testing criteria for the kinetic study under the respirometric conditions. PMID- 12906301 TI - Effect of nitrite and nitrate on biogenic sulfide production in sewer biofilms determined by the use of microelectrodes. AB - The effects of O2 and NO3- concentrations on in situ sulfate reduction and sulfide reoxidation in microaerophilic wastewater biofilms grown on rotating disk reactors were investigated by the use of microelectrodes for O2, S2-, NO3-, NO2-, and pH. Microelectrode measurements showed the vertical microzonation of O2 respiration, NO3- respiration, H2S oxidation and SO4(2-) reduction in the biofilms. The microelectrode measurements indicate that sulfate reducing activity was largely restricted to a narrow anaerobic zone located about 500 microm below the biofilm surface. An addition of nitrate forced the sulfate reduction zone deeper in the biofilm and reduced the specific sulfate reduction rate as well. The sulfate reduction zone was consequently separated from the O2 and NO3- respiration zones. Anaerobic H2S oxidation with NO3- was also induced by addition of nitrate to the medium. Measurements of the reduced inorganic sulfur compounds (FeS, FeS2 and S0), total-Mn and total-Fe in the biofilm indicated that the produced H2S became immediately oxidized with O2, NO3- and other oxidants, mainly ferric/ferrous hydrates. On the basis of the present results, it was estimated that of all sulfide produced, 13% of the sulfide was precipitated by metal ions as FeS and S0 just above the sulfate reduction zone, 65% was anaerobically oxidized to SO4(2-) with NO3- as an electron acceptor and 22% was aerobically oxidized within the biofilm incubated in 70 micromol l(-1) of DO and 280 micromol l(-1) of NO3-. PMID- 12906302 TI - Non-filamentous sludge bulking caused by a deficiency of nitrogen in industrial wastewater treatment. AB - Deficiency in the nutrient supply such as nitrogen usually results in activated sludge bulking and this phenomenon often takes place in the industrial wastewater treatment plants with activated sludge process. The effects of nitrogen deficiency on activated sludge bulking were studied specially in some experiments carried out in a sequencing batch reactor fed with brewing process wastewater in this paper. The experimental results showed that the sludge settled properly at an influent BOD/N value of 100/4. When the value of BOD/N was 100/3, filaments had an excessive growth at one time during the reaction process. Afterwards, the number of filamentous bacteria began to reduce and simultaneously an excessive growth of viscous Zoogloea with high percentage of moisture was observed and non filamentous activated sludge bulking occurred. When the influent BOD/N value was 100/2, the excessive growth of filamentous microorganisms could not be observed at all times and the sludge characterization was similar to the case in which BOD/N value was 100/3. When the value of influent BOD/N was 100/0.94, more serious non-filamentous bulking occurred. Furthermore, the effects of nitrogen deficiency on the nitrogen sources and phosphorus sources utilization rate and the COD removal rate were investigated in the experiments. PMID- 12906303 TI - Oxygen transfer into activated sludge with high MLSS concentrations. AB - In this report, tests on the impact of the sludge properties on the oxygen transfer at low and high solids contents are presented. Additional to the oxygen transfer tests, the activated sludge was intensively analysed to examine the changes of the alpha-factor in relation to the sludge properties (rheology, EPS, CST, etc.). The alpha-factor did strongly decrease in all sludge types at increasing MLSS or increasing viscosity, respectively. In the second test stage, the impact of the aeration system was examined in detail. For these tests, the same sludge from a membrane bioreactor was used throughout. Apart from the impact of the power density in the reactor and the specific air throughput, the main focus was on the economic efficiency of the examined systems in cases of high MLSS. It became apparent that up to solids contents of 18 g/l the fine-bubble aeration is the most economically efficient method. PMID- 12906304 TI - Optimum aerobic volume control based on continuous in-line oxygen uptake monitoring. AB - Dynamic adaptation of the aerated volume to changing load conditions is essential to maximise the nitrogen removal performance and to minimise energy consumption. A control strategy is presented which provides optimum aerobic volume control (OAV-control concept) based on continuous in-line oxygen uptake monitoring. For ammonium concentrations below 1 mg/l the oxygen uptake rate shows a strong and almost linear dependency on the ammonium concentration. Therefore, the oxygen uptake rate is an ideal indicator for the nitrification performance in activated sludge systems. The OAV-control concept provides dynamic variation of the minimum aerobic volume required for complete nitrification and therefore maximises the denitrification performance. In-line oxygen uptake monitoring is carried out by controlling the oxygen concentration in a continuous aerated zone of the aeration tank and measuring the total air flow to the aeration tank. The total air flow to the aeration tank is directly proportional to the current oxygen uptake rate and can therefore be used as an indicator for the required aerobic volume. The instrumentation requirements for installation of the OAV-control are relatively low, oxygen sensors in the aeration tank and an on-line air flow measurement are needed. This enables individual control of aeration tanks operated in parallel at low investment costs. The OAV-control concept is installed at the WWTP Linz-Asten (1 Mio PE) and shows very good results. Full scale results are presented. PMID- 12906305 TI - Investigation of oxygen transfer rates in full scale membrane bioreactors. AB - In membrane bioreactors (MBRs) for wastewater treatment the secondary clarifier is replaced by a membrane filtration. The advantage of this process is a complete removal of solids from the effluent and a small footprint due to possible high biomass concentrations (MLSS). As oxygen supply counts for more than 70% of total energy cost in municipal WWTPs the design of the aeration system is vital for efficient operation. In this respect the alpha-value is an important influencing factor. The alpha-value depends on the MLSS-concentration as shown in various publications and confirmed by own measurements in two full scale municipal MBRs with MLSS ranging from 7 and 17 kg/m3. Furthermore it must be taken into account that alpha-values are not static values; they vary with loading rates, surfactant concentrations, air flow rates, MLSS concentrations, etc. The average alpha-value at typical 12 kg/m3 MLSS for municipal MBRs is about 0.6 +/- 0.1. As submerged configured MBRs are equipped with an additional coarse bubble "crossflow" aeration system for fouling control, supplementary energy is consumed. Therefore MBRs need more energy compared to conventional treatment plants. Measurements of both aeration systems show that the fine bubble aeration system is more efficient by a factor of three concerning oxygen supply compared to the coarse bubble system. PMID- 12906306 TI - Analysis of polyethylene-glycol-polylactide nano-dimension artificial red blood cells in maintaining systemic hemoglobin levels and prevention of methemoglobin formation. AB - We have recently reported our study on novel nano-dimension red blood cell (rbc) substitute based on ultrathin PEG-PLA membrane nanocapsules (80-150 nanometer diameter) containing hemoglobin (Hb) and enzymes. These have a markedly increased the circulation half-times as compared to our earlier PLA membrane nanocapsules. In the present study to be reported here, instead of looking at this from a pharmacodynamic point of view, we design the Hb nanocapsules from the point of view of transfusion medicine. For instance, the maximal levels of systemic non red blood cell (rbc) Hb that can be attained after one infusion of 30% blood volume of 10 gm/dl Hb in the form of different types of PEG-PLA Hb nanocapsules or polyHb. Also the length of time one infusion can maintain a given systemic non rbc hemoglobin Hb level. Of the two types of polyhemoglobins similar to those in clinical trials but prepared in this laboratory, the maximal levels of Hb reached were 3.35 gm/dl and 3.10 gm/dl respectively. The times for the hemoglobin level to fall to 1.67 gm/dl were 14 hours and 10. hours respectively, corresponding to 24 hours and 17 hours in human. The best PEG-PLA Hb nanocapsules are prepared using a combination of the following 4 factors: use of polymerized Hb, the use of higher M.W. PLA, the use of higher concentrations of PEG-PLA and the crosslinking of the newly formed PEG-PLA Hb nanocapsules. With this, the maximal non-rbc systemic Hb reached was 3.66 gm/dl and the time to reach 1.67 gm/dl was 24.2 hours, or 41.5 hours in human if extrapolated using the results obtained with polyHb in rats. PMID- 12906307 TI - Polypropylene hollow fiber for cells isolation: methods for evaluation of diffusive transport and quality of cells encapsulation. AB - Formulation of membrane properties is important prior the successful implantation of encapsulated cells producing therapeutically relevant compounds. The purpose of our study was to specify the methods allowing preliminary evaluation of hollow fibers (HF) chosen for immunoisolation. We have selected as estimates (1) diffusive permeability for small and large solutes, and HF cut off (in vitro), (2) histological evaluation of tissue overgrowth after sc. implantation into mice. It was found that diffusive coefficients were linearly dependent on the particle diameter except that of albumin (2-3 times higher than theoretically estimated). This discrepancy imply that for certain particles the interaction with membrane material may be significant. The histological evaluation showed that siliconized HF implanted for 105 days were accepted (there was thin fibrotic layer on the external surface of the HF, no surrounding haemopoietic cells were found). It is concluded that proposed methods for preliminary evaluation of hollow fibers chosen for immunoisolation seems to be reliable and suitable for testing diffusive permeability of each relevant cell product. PMID- 12906308 TI - Osteoblasts cultured from osteoporotic bone: a comparative investigation on human and animal-derived cells. AB - In vitro studies on pathophysiology and innovative treatments of many orthopaedic diseases, based on the investigations of cells from pathologic skeletal tissues, greatly improve basic knowledge of osteoporosis. Primary osteoblast (OB) cultures derived from osteopenic bone from different species (human, rat, sheep) were compared to assess the differences that should be taken into account when performing in vitro biocompatibiliy tests or investigating pharmacological and physical treatments. Primary OB were isolated from osteopenic patients and animals by well-established methods and their metabolism was assessed with or without 1,25(OH)2D3. The greatest significant differences were observed between rat and human cells both under basal conditions and after 1,25(OH)2D3 stimulation. In addition, the response to 1,25(OH)2D3 stimulation of OBs from osteopenic rats was significantly different from that of human and sheep OB cultures, in terms of NO, OC, IL-6, and TGF-beta1. Cells derived from osteopenic sheep behaved much more similarly to those from humans, except for a significant difference in terms of TGF-beta1 observed both under basal conditions and after stimulation. PMID- 12906309 TI - The preparation and characterization of monomethoxypoly(ethylene glycol)-b-poly DL-lactide microcapsules containing bovine hemoglobin. AB - Methoxypoly(ethylene glycol)-b-poly-DL-lactide (PELA) microcapsules containing bovine hemoglobin (bHb) were prepared by a W/O/W double emulsion-solvent diffusion process. bHb solution was used as the internal aqueous phase, PELA/organic solvent as the oil phase, and polyvinyl alcohol (PVA) solution as the external aqueous phase. This W/O/W double emulsion was added into a large volume of water (solidification solution) to allow organic solvent to diffuse into water. The optimum preparative condition for PELA microcapsules loaded with bovine hemoglobin was investigated. It was found that homogenization rate, type of organic solvent, and volume of the solidification solution influenced the activity of bovine hemoglobin encapsulated. When the homogenization rate was lower than 9000 rpm and ethyl acetate was used as the organic solvent, there was no significant influence on the activity of hemoglobin. High homogenization rate as 12 000 rpm decreased the P50 and Hill coefficient. Increasing the volume of solidification solution had an effect of improving the activity of microencapsulated hemoglobin. The composition of the PELA had the most important influence on the success of encapsulation. Microcapsules fabricated by PELA with MPEG2k block (molecular weight of MPEG block: 2000) achieved a high entrapment efficiency of 90%, better than PL A homopolymer and PELA with MPEG5k blocks. Hemoglobin microcapsules with native loading oxygen activity (P50 = 26.0 mmHg, Hill coefficient = 2.4), mean size of about 10 microm, and high entrapment efficiency (ca. 93%) were obtained at the optimum condition. PMID- 12906310 TI - Studies on the preparation of chitosan microcarriers cross-linked by oxidized lactose and culture of primary hepatocytes. AB - Chitosan microcarriers were prepared using oxidized lactose as a cross-linking agent which was oxidized by sodium periodate. The effect of amount of oxidized lactose on the preparation of chitosan microcarriers was studied and optimized. Rat hepatocytes cultivated on chitosan microcarriers cross-linked by oxidized lactose retained the spherical shape as they have in vivo. Liver-specific functions such as albumin secretion and glucose metabolism were stably maintained for seven days. The metabolic activities of hepatocytes cultured on the oxidized lactose cross-linked chitosan microcarriers were higher than those of hepatocytes on chitosan microcarriers cross-linked by glutaraldehyde and on cytodex 3. The results suggest that oxidized lactose could be an interesting cross-linking agent for chitosan thus reducing the toxic side effects caused by using glutaraldehyde. PMID- 12906311 TI - Encapsulating protein into preformed liposomes by ethanol-destabilized method. AB - This study describes a highly efficient method for encapsulating protein drugs into liposomes without using toxic solvents such as chloroform. Large unilamellar vesicles (LUVs) were formed by the ethanol injection method. The effects of composition of phospholipid, buffer concentration, incubation time, incubation temperature, drug loading, ethanol content, and the presence of poly(ethylene) glycol (PEG) lipids on the entrapment efficiency of protein were investigated. It was shown that these preformed LUVs could be induced to entrap protein drugs in the presence of ethanol. Protein could be efficiently encapsulated into liposomes. The interaction of the liposomes with proteins leads to the formation of multilamellar liposomes ranging in size from 70 to 120 nm, only slightly bigger than the parent LUVs from which they originated. Protein drugs were stable in the liposomal solution. There is no significant activity loss during the encapsulation process. The optimal encapsulation efficiency was achieved when 30% approximately 40% ethanol was used in encapsulating protein drugs. Due to the steric hindrance, LUVs containing a PEG coating will dramatically reduce the encapsulation efficiency, even in liposomes containing very low amount of PEG. PMID- 12906312 TI - Continuous production of cephalosporin-C by immobilized microbial cells using symbiotic mode in a packed bed bioreactor. AB - Cephalosporins are usually produced semisynthetically from Cephalosporin-C, which is exclusively produced by Cephalosporium acremonium. Free cell studies for the production of Cephalosporin-C had some limitation such as pulpy growth of fungus causing an appreciable rise in the broth viscosity affecting the transfer of oxygen and other nutrients into the cells. High cell concentrations cannot be maintained because of wash out phenomenon at high dilution rates. The whole cell immobilization technique is a potentially important process for Cephalosporin-C biosynthesis, where increase cell densities were maintained and broth-handling problems were reduced. Cephalosporin-C fermentation is a highly aerobic process. The symbiotic relationship of Cephalosporium acremonium and Chlorella pyrenoidosa has been used to increase oxygen transfer rate to the fungi by co-immobilizing it with algae. Co-immobilization of whole cells of fungus and algae were carried out in different immobilizing agents and the systems were coated with polyacrylamide resin of pharmaceutical grade to overcome the problems of leakage. The operational stability of immobilized systems in a packed bed reactor was also studied. PMID- 12906313 TI - An enzyme electrode based on lipoxygenase immobilized in gelatin for selective determination of essential fatty acids. AB - An enzyme electrode for the specific determination of omega-3 and omega-6 fatty acids from the mixture of essential fatty acids (EFAs) was developed by using lipoxygenase (LOX) (EC 1.13.11.12) from soy beans in combination with a dissolved oxygen (DO) probe. The enzyme electrode showed different sensitivities for linoleic (LA) and alpha-linolenic acids (ALA), the most common essential fatty acids. Enzyme electrode response depends linearly on LA concentration between 12.8-160.5 microM and ALA concentration between 3.8-18.9 microM in borate buffer, 0.2 M at pH 9.0. However, in phosphate buffer 0.2 M at pH 6.0 linearity is in the range of 7.5-22.5 microM of ALA concentration at 5 minutes response times. Moreover, maximum electrode response was found in borate buffer at pH 9.0 and 30 degrees C. PMID- 12906314 TI - Preparation and properties of alpha-galactosidase chemically attached to activated chitin. AB - alpha-Galactosidase (alpha-D-galactoside galactohydrolase, EC 3.2.1.22) from watermelon was covalently immobilized on chitin. The immobilized alpha galactosidase exhibited an activity of 0.61 U per g of carrier and an activity yield of 67%. The properties of free and immobilized alpha-galactosidase were also searched and compared. The results showed that, optimum conditions for activity were not affected by immobilization. The optimum pH and temperature for free and immobilized enzyme found as pH 6.0 and 65 degress C, respectively. Compared with the free enzyme, the temperature and pH stabilities of the immobilized enzyme were similar. Both the enzymes were stable between pH 2-10 and below 50 degrees C. The Km values for free and immobilized enzyme were determined using p-nitrophenyl-alpha-D-galactopyranoside (PNPG) and raffinose as substrates. Operational stability of the immobilized enzyme was investigated by using both substrates. The operational half-life (t 1/2) was calculated as 34 h for PNPG and 28 h for raffinose. The immobilized alpha-galactosidase was also utilized in the hydrolysis of raffinose. The immobilization procedure on chitin was cheap and also easy to carry out, and the immobilized enzyme had good properties that the potential for practical application is considerable. PMID- 12906315 TI - Arginine selective biosensor based on arginase-urease immobilized in gelatin. AB - Arginase and urease enzymes were immobilized on the surface of pH electrode by using gelatin membrane which is then cross-linked with glutaraldehyde. Sensor response was maximum when 2.5 mM, Tris-HCl buffer (pH 8.5) was used at 25 degrees C. The biosensor response depends linearly on arginine concentration between 0.025-0.310 mM with response time 10 min. Furthermore, application of the system for the arginine detection in serum samples was also tested. PMID- 12906316 TI - Angiogenesis in psoriasis. PMID- 12906317 TI - Anti-angiogenic therapy: rationale, challenges and clinical studies. AB - Physiological angiogenesis occurs during embryogenesis, wound healing and reproductive functions in adults. Abnormal angiogenesis takes place in certain chronic diseases (diabetes, psoriasis, rheumatoid arthritis, etc.) and tumours. Genetic changes and local stresses including hypoxia, glucose deprivation and oxidative stress play a pivotal role in angiogenesis switch, which is necessary for tumour development and is rate-limiting for tumour progression. Angiogenesis is tightly regulated by pro- and anti-angiogenic growth factors with a series of complex and interrelated steps. Activated endothelial cells (ECs) migrate as a solid cord and, subsequently, form lumina; the sprout tips then anastomose to form vessel loops or networks. One of the final events is the laying down of a basement membrane and the structural support of pericytes. The molecular alterations that sustain angiogenesis represent novel targets for rationally designed anti-cancer treatment strategies. Inhibition of angiogenesis presents certain advantages on conventional therapies, such as the direct accessibility from the circulation, and the potential low rate of drug resistance related to the genetic stability of ECs. Certain anti-angiogenic compounds were found to have potent anticancer property in in vivo experimental studies. Nevertheless, in contrast to preclinical studies, the first generation of anti-angiogenic drugs tested in clinical trials have shown a moderate activity in advanced disease partly due to suboptimal schedules of therapy or biases in study design. PMID- 12906318 TI - Importance of vascular endothelial growth factor A in the progression of experimental neuroblastoma. AB - Vascular endothelial growth factor A (VEGF-A) and its receptor tyrosine kinases located on endothelial cells seem to play an important role in the multistep pathway of angiogenesis. SU5416 is a small molecule which inhibits angiogenesis by acting as an inhibitor of VEGF receptor-2 tyrosine kinase. We have developed a reproducible murine model for neuroblastoma, a childhood cancer, based on s.c. xenotransplantation of SH-SY5Y neuroblastoma cells. We found that SH-SY5Y cells expressed VEGF-A on both the mRNA and protein levels, that plasma concentrations of VEGF-A were significantly elevated in animals with neuroblastoma with a volume > 1.4 ml, and that there was a correlation between VEGF-A levels in plasma and tumor size in untreated tumor-bearing animals. Treatment with SU5416 reduced the growth of neuroblastoma tumors by 65% without apparent toxicity. SU5416 treatment also suppressed tumor angiogenesis, despite an increase in plasma VEGF-A levels per ml tumor volume during therapy. Our experimental data suggest that the angiogenesis inhibitor SU5416 may be beneficial in the treatment of solid tumors of childhood such as neuroblastoma. PMID- 12906319 TI - Atypical cystic fibrosis--diagnostic and management dilemmas. PMID- 12906321 TI - Posture and cystic fibrosis. PMID- 12906320 TI - Autonomic dysfunction in cystic fibrosis. PMID- 12906322 TI - Exercise testing in children with cystic fibrosis. PMID- 12906323 TI - Asthma in cystic fibrosis. PMID- 12906324 TI - Growth and growth charts in cystic fibrosis. PMID- 12906325 TI - Coping with cystic fibrosis. PMID- 12906326 TI - The daily grind and how to stay sane as a mother of two children with cystic fibrosis. PMID- 12906327 TI - Sequencing of the Pseudomonas aeruginosa and Burkholderia cepacia genomes and their applications in relation to cystic fibrosis. PMID- 12906329 TI - Double liposomes: hypoglycemic effects of liposomal insulin on normal rats. AB - The biopharmaceutical characteristics of double liposomes (DLs) containing insulin were examined, and the usefulness of DLs in combination with aprotinin is discussed. Encapsulation of insulin was influenced by lipid composition, and the highest efficiency was observed with positively charged liposomes. Insulin encapsulated in liposomes, especially in DLs, was protected from enzymatic proteolysis. A portion of insulin molecules was adsorbed on the surface of the membrane when liposomes were prepared using a lipid with a positive charge and was degraded by enzymes. Remarkable hypoglycemic effects were observed after intragastric administration of DLs containing insulin at a dose of 20 IU/kg to normal male Wistar rats. The highest mean relative efficacy to administration was obtained with insulin-loading DLs containing aprotinin as a protease inhibitor. These results suggest that DLs are applicable as an oral dosage form for peptide drugs such as insulin etc., especially in combination with protease inhibitors. PMID- 12906328 TI - Recent advances in cross-infection in cystic fibrosis: Burkholderia cepacia complex, Pseudomonas aeruginosa, MRSA and Pandoraea spp. PMID- 12906330 TI - Behavior of decomposition of rifampicin in the presence of isoniazid in the pH range 1-3. AB - The extent of decomposition of rifampicin in the presence of isoniazid was determined in the pH range 1-3 at 37 degrees C in 50 min, the mean stomach residence time. With increase in pH, the degradation initially increased from pH 1 to 2 and then decreased, resulting in a bell-shaped pH-decomposition profile. This showed that rifampicin degraded in the presence of isoniazid to a higher extent at pH 2, the maximum pH in the fasting condition, under which antituberculosis fixed-dose combination (FDC) products are administered. At this pH and in 50 min, rifampicin decomposed by approximately 34%, while the fall of isoniazid was 10%. The extent of decomposition for the two drugs was also determined in marketed formulations, and the values ranged between 13-35% and 4 11%, respectively. The extents of decomposition at stomach residence times of 15 min and 3 h were 11.94% and 62.57%, respectively, for rifampicin and 4.78% and 11.12%, respectively, for isoniazid. The results show that quite an extensive loss of rifampicin and isoniazid can occur as a result of interaction between them in fasting pH conditions. This emphasizes that antituberculosis FDC formulations, which contain both drugs, should be designed in a manner that the interaction of the two drugs is prevented when the formulations are administered on an empty stomach. PMID- 12906331 TI - Preparation and characterization of glassy celecoxib. AB - Celecoxib, a poorly water-soluble drug, was converted into a glassy state by melt quenching. The properties of glassy celecoxib were studied using infrared (IR) spectroscopy, differential scanning calorimetry (DSC), X-ray powder diffraction (XRPD), intrinsic dissolution rate (IDR), and thin-layer-chromatography (TLC). Glass transition occurred at 51.8 degrees C. Infrared spectrum of glass has revealed significant changes due to H-bonding. Celecoxib glass shows around 15 times faster dissolution as compared with the crystalline state. Heckel plot analysis has shown better compressibility in glassy state. Unpulverized glass remained stable for 3 months, whereas after pulverization about 70% crystallinity was gained in 100 hours. Further attempts may be carried out to stabilize the glass. PMID- 12906332 TI - Evaluation of PLGA microspheres as delivery system for antitumor agent camptothecin. AB - Camptothecin (CPT) and its analogues are a new class of anticancer agents that have been identified over the past several years. Camptothecin exists in two forms depending on the pH: An active lactone form at pH below 5 and an inactive carboxylate form at basic or physiological neutral pH. Poly(lactide-co-glycolide) (PLGA) microspheres have been considered good delivery vehicles for CPT because of acidic microenvironment formed through PLGA degradation. The objective of this study is to investigate antitumor activity of CPT after it is encapsulated in PLGA microspheres. In this study, PLGA microspheres containing various CPT loadings were prepared and characterized. Cytotoxicity of these microspheres to B16 melanoma cells was then evaluated, and uptake of microspheres by B16 cells was also studied. Analysis of drug stability revealed that CPT is released from the microspheres in its active lactone form over the entire release duration. It was also found that there was no interaction between CPT and PLGA matrix within microspheres through Differential Scanning Calorimetry (DSC) and Fourien Transform Infrared Spectroscopy (FT-IR) and hign performance liquid chromatography (HPLC) studies. Cytotoxicity assay showed that CPT encapsulated in PLGA microspheres still retained its antitumor potency. Uptake study revealed quick uptake of the microspheres by B16 cells, which was desirable. It was concluded that PLGA microspheres were suitable delivery vehicles to stabilize and deliver CPT for the treatment of cancer. PMID- 12906333 TI - Production and characterization of hot-melt extruded films containing clotrimazole. AB - Hot-melt extrusion technology (HME) was used to prepare muco-adhesive matrix films containing 10% w/w clotrimazole (CT) intended for local drug delivery applications for the oral cavity. This study was aimed at the production and characterization of these drug delivery systems for the prophylaxis and treatment of oral candidiasis. The film system's formulation contained hydroxypropyl cellulose and poly(ethylene oxide) as polymeric carriers, the bioadhesive polycarbophil, and other excipients. The CT formulation was processed at a temperature range of 125-130 degrees C utilizing a Killion extruder (Model KLB 100) equipped with a 6-inch flex-lip die. The films were evaluated for postextrusion drug content, physical and chemical content uniformity, drug release, thermal and crystalline behavior, and bioadhesive strength. The extruded films demonstrated excellent content uniformity and a postprocessing drug content of 93.3% (+/- 1.0). The degradation product, (o-chlorophenyl)diphenyl methanol, was also identified and quantitated using high performance liquid chromatography. The films were determined to exhibit desirable and consistent release properties and bioadhesive strength (p < 0.05). The results of this study indicate that HME is a viable technique for the preparation of muco-adhesive films containing clotrimazole for oral candidiasis. PMID- 12906334 TI - Effect of sodium alterations on hepatic cytochrome P450 3A2 and 2C11 and renal function in rats. AB - Numerous dietary supplements are known to modulate cytochrome P450 (CYP)-mediated metabolism and subsequently alter drug toxicity or efficacy in animals and humans. In the present study we investigated the effect of varying amounts of sodium intake on renal function and the metabolic activity of the hepatic CYP3A2 and CYP2C11 isoforms. Rats were maintained on standard rodent chow or a low-salt rice diet. Within each of these groups rats received either a single intraperitoneal injection of furosemide to initiate salt depletion, or saline. Additional groups included salt supplementation of 500 mg/300 g body weight/day and 1.25 g/300 g body weight/day of sodium chloride solution. Rats receiving the low-salt diet, both with and without a concomitant furosemide administration, had a significant reduction in creatinine clearance without changes in serum creatinine. In addition, urine flow rate was markedly reduced in rats maintained on the low-salt diet. Western blot analysis indicated that neither sodium supplementation nor deprivation altered hepatic microsomal CYP3A2 levels; however, hepatic CYP2C11 levels significantly increased in rats receiving the largest sodium supplement. In vitro metabolic activity of CYP3A2 was unchanged as compared with controls. Activity of CYP2C11 was significantly reduced in both rat groups receiving additional sodium supplements. Acute manipulation of daily sodium intake does alter renal function and specific hepatic CYP isoforms and should be considered when using these rat models. PMID- 12906335 TI - Comparison of dissolution profiles for albendazole tablets using USP apparatus 2 and 4. AB - The in vitro dissolution of albendazole from three different commercially available products (200 mg tablets) was studied using U.S. Pharmacopeia (USP) Apparatus 2 and USP Apparatus 4 in order to compare the release performance of the drug in two essentially different dissolution systems. For both cases, 0.1 N HCl was used as dissolution medium. Only the reference product and one of the generic products studied met the 80% USP 24 specification for albendazole dissolved at 30 min, using USP Apparatus 2. Although the reference product reached 80% of albendazole dissolved at 30 min when Apparatus 4 was used, the generic products' dissolution performance was markedly reduced in this system. Though dissolution rate was slower using Apparatus 4, the total quantity of albendazole dissolved from the reference product, represented by area under the dissolution profile, was practically the same regardless of the system used. Dissolution kinetics of albendazole was adequately described by Weibull's function for all the products. The dissolution time (t(d)) derived from data fitting to this function showed significant differences among the products studied. Data analysis based on analysis of variance (ANOVA) showed nonequivalence among the dissolution profiles of generic products compared with the reference product either with the dissolution vessel system or the flow through cell, as well as nonequivalence among the dissolution profiles using both apparatuses with the same product. Though differences in the dissolution profiles for generic products against the reference product in both systems were found, USP Apparatus 4 showed higher discriminative capacity in differentiating the release characteristics of the products tested. PMID- 12906336 TI - Development of matrix patches for transdermal delivery of a highly lipophilic antiestrogen. AB - The aim of this study was to develop matrix-type transdermal systems (TDSs) containing the highly lipophilic (log P = 5.82) antiestrogen (AE) and the permeation enhancers propylene glycol and lauric acid. For that purpose, permeation of AE from various adhesive matrices through excised skin of hairless mice was evaluated. It was found that pretreatment of the skin with permeation enhancers raised the transdermal flux of subsequently applied antiestrogen. Highest steady-state transdermal fluxes (1.1 microg cm(-2) h(-1)) were obtained from Gelva, polyacrylate adhesive, followed by 0.55 microg cm(-2) h(-1) from Oppanol polyisobutylene, 0.31 microg cm(-2) h(-1) from BIO-PSA silicone, and 0.12 microg cm(-2) h(-1) from Sekisui polyacrylate matrices. In order to develop TDS with high content of fluid permeation enhancer propylene glycol, two different strategies were investigated. One strategy was the addition of hydroxypropyl cellulose (HPC) as thickening agent to Gelva matrices. This allowed for propylene glycol loading levels of up to 30%, resulting in transdermal AE fluxes of 0.09 microg cm(-2) h(-1). On the other hand, a fleece-laminated backing foil was loaded with the described permeation enhancer formulation and laminated with polyacrylate adhesive layer, resulting in transdermal AE fluxes of 0.06 microg cm(-2) h(-1). However, application of these TDSs on skin pretreated with permeation enhancers raised the fluxes to 2.6 microg cm(-2) h(-1) from Gelva/HPC and 0.46 microg cm(-2) h(-1) from fleece/Sekisui. PMID- 12906338 TI - Solid-state variation of troglitazone drug substance by using a different recrystallization method. AB - Rapid and slow crystallization methods (A and B) were applied for troglitazone, an equal mixture of four stereoisomers. Differences in the powder x-ray diffractometry patterns and hygroscopic patterns were observed among the samples crystallized by these methods, suggesting that troglitazone has solid-state variation. In this article, troglitazone recrystallized by method A was evaluated to clarify its structural characteristics and physical property. The crystal structure of predried troglitazone recrystallized by method A was proved to be a dihydrate. By drying, it changed reversibly to an anhydrate, which is the same structure as the RS/SR form, keeping the same enantiomer ratio. The solubility of the troglitazone by method A higher than that by method B at all enantiomer levels. But making the troglitazone amorphous equalized the enantiomeric solubilities of the substances by both methods as well as increased the intrinsic solubilities. Troglitazone by both methods was proved to be stable and retained the ratio of the stereoisomers. PMID- 12906337 TI - Release profiles of theophylline from microspheres consisting of dextran derivatives and cellulose acetate butyrate: effect of polyion complex formation. AB - The objective of this study was to evaluate the utility of mixtures among oppositely charged dextran derivatives as constituents of a controlled release microsphere. Carboxymethyldextran (CMD) and dextran sulfate (DS) were used as polyanions, and [2-(diethylamino) ethyl] dextran (EA) and [2 hydroxypropyltrimethylammonium] dextran (CDC) as polycations. The microspheres consisting of hydrophilic and hydrophobic polymers were prepared by emulsion solvent evaporation method. The mixtures, CMD/EA, CMD/CDC, DS/EA, and DS/CDC, were used as hydrophilic polymers, because they can interact with each other to form polyion complexes for the improvement of sustained-release performances. Cellulose acetate butyrate and theophylline were used as a model hydrophobic polymer and a model drug, respectively. The yield of microspheres was excellent (more than 95%). According to observation, by scanning election microscopy (SEM) microspheres were spherical with a rough surface. The in vitro drug release from microspheres was examined in the JP XIV first fluid, pH 1.2, and second fluid, p H 6.8, at 37 degrees C, and 100 rpm. In the DS/CDC system, drug release was depressed by formation of a polyion complex and not affected by pH of dissolution medium. The release rate was modulated by the ratio of hydrophilic and hydrophobic matrix. This particulate system, in which the polyion complex matrix is strengthened by a hydrophobic polymer, is a promising formulation for drug delivery. PMID- 12906339 TI - Evaluation of monolithic osmotic tablet system for nifedipine delivery in vitro and in vivo. AB - The aim of this study was to evaluate the monolithic osmotic tablet system (MOTS) containing a solid dispersion with the practically water-insoluble drug nifedipine in vitro and in vivo. In the drug release study in vitro, the release profiles of this system had almost zero-order kinetics. The influences of tablet formulation variables, sizes of the delivery orifice, membrane variables, and values of pH in the dissolution medium on nifedipine release from MOTS have been investigated. The results provided evidence that the tablet core played an important role in MOTS. While orifice sizes and membrane variables affected the nifedipine release rate, MOTS was independent of the dissolution medium. The appropriate orifice size was found to be in the range of 0.5-1.0 mm. The coating membrane incorporating hydrophilic polyethylene glycol (PEG) formed a porous structure. The human pharmacokinetics and relative bioavailability of MOTS containing nifedipine were compared with a commercial Adalat osmotic tablet system containing an equivalent dose of nifedipine following an oral single dose of 30 mg given to each of 11 healthy volunteers in an open, randomized crossover study in vivo. The relative bioavailability for MOTS was 112%. There was no statistically significant difference in the pharmacokinetic parameters between two dosage forms. It is concluded that the monolithic osmotic tablet controlled release system is feasible for a long-acting preparation as a once-daily treatment. PMID- 12906340 TI - Pediatric mood and anxiety syndromes in primary care: who gets identified?. AB - OBJECTIVE: To seek clues to the enhancement of primary care management by (i) Determining how often and in whom primary care clinicians in the United States, Puerto Rico, and Canada identify pediatric mood or anxiety syndromes; (ii) Determining which clinical and demographic features predict higher rates of identification; (iii) Describing assessment methods used. METHODS: This report uses the database of the multi-site Child Behavior Study. This cross-sectional study involved 206 primary care practices in the United States, Puerto Rico, and Canada; 395 clinicians and 20,861 primary care attenders aged 4-15 years. Clinicians completed a visit questionnaire addressing presence and type of psychosocial problems and how assessed. Parents completed a questionnaire addressing family demographics, child symptoms (Pediatric Symptom Checklist) and functioning, and child service use. RESULTS: Clinicians identified psychosocial problems on 17.9% of visits, but mood or anxiety syndromes on only 3.3%, most commonly in children judged to have co-morbid behavioral syndromes, of whom the majority (66.7%) already had contact with specialized mental health. Neither parental concerns about mood and anxiety symptoms nor clinician familiarity with the patient were major predictors of identification. When making a diagnosis of a pure internalizing syndrome (i.e., without a co-morbid behavioral syndrome) clinicians rarely used standardized tools or school reports. CONCLUSIONS: Neither screening for nor diagnosis of mood and anxiety syndromes is a routine part of primary care of children and adolescents. Efforts to improve care must include practical, validated screening procedures to enhance assessment for mood and anxiety syndromes, particularly among children in whom primary care clinicians identify psychosocial problems. PMID- 12906341 TI - Integrating clinical nurse specialists into the treatment of primary care patients with depression. AB - OBJECTIVE: To examine the effectiveness of integrating generalist and specialist care for veterans with depression. METHOD: We conducted a randomized trial of patients screening positive for depression at two Veterans Affairs Medical Center general medicine clinic firms. Control firm physicians were notified prior to the encounter when eligible patients had PRIME-MD depression diagnoses. In the intervention firm, a mental health clinical nurse specialist (CNS) was to: design a treatment plan; implement that plan with the primary care physician; and monitor patients via telephone or visits at two weeks, one month and two months. Primary outcomes (depressive symptoms, patient satisfaction with health care) were collected at 3 and 12 months. RESULTS: Of 268 randomized patients, 246 (92%) and 222 (83%) completed 3- and 12-month follow-up interviews. There were no between-group differences in depressive symptoms or satisfaction at 3 or 12 months. The intervention group had greater chart documentation of depression at baseline (63% versus 33%, p = 0.003) and a higher referral rate to mental health services at 3 months (27% versus 9%, p = 0.019). There was no difference in the rate of new prescriptions for, or adequate dosing of, anti-depressant medications. In 40% of patients, CNSs disagreed with the PRIME-MD depression diagnosis, and their rates of watchful waiting were correspondingly high. CONCLUSIONS: Implementing an integrated care model did not occur as intended. Experienced CNSs often did not see the need for treatment in many primary care patients identified by the PRIME-MD. Integrating integrated care models in actual practice may prove challenging. PMID- 12906342 TI - PTSD, depression, and health status in Lebanese civilians exposed to a church explosion. AB - OBJECTIVE: This study examined the effect of stressors on PTSD, depression, and health status in a sample of Lebanese civilians exposed to a church explosion and their comparison groups (33 victims, 30 family members, and 30 neighbors) one year after the event. METHOD: This descriptive study used interviews that were conducted with the participants in their homes. The main outcome measures were the Clinician Administered PTSD Scale Version I, Beck Depression Inventory, and change in health service utilization in the past year. Multivariate analysis was done using logistic regression to predict the study outcomes. RESULTS: PTSD was present in 17.2% of the total sample, depression in 41.9% and the mean increase in doctors' visits was 2.05 +/- 2.97. In the victims' group, 39% met PTSD diagnostic criteria, 51% were depressed, and 45% reported deterioration in their health status. These rates were significantly higher than those in the comparison groups. Also, females were 2.62 times more at risk than males for depression. Victims were 7.35 times and those with financial problems 2.67 times more at risk of having increased doctor's visits than their family or neighbor comparison groups. CONCLUSION: The results suggest that PTSD, depression, and health status deterioration in trauma victims may persist over time. Resources such as social support can help attenuate the symptoms of those at a higher risk. PMID- 12906343 TI - Psychological burden in the era of HAART: impact of selenium therapy. AB - OBJECTIVE: To determine the impact of nutritional (selenium) chemoprevention on levels of psychological burden (anxiety, depression, and mood state) in HIV/AIDS. METHOD: A randomized, double-blind, placebo-controlled selenium therapy (200 microg/day) trial was conducted in HIV+ drug users from 1998-2000. Psychosocial measures (STAI-State and Trait anxiety, BDI-depression, and POMS- mood state), clinical status (CD4 cell count, viral load), and plasma selenium levels were determined at baseline and compared with measurements obtained at the 12-month evaluation in 63 participants (32 men, 31 women). RESULTS: The majority of the study participants reported elevated levels of both State (68%) and Trait (70%) anxiety. Approximately 25% reported overall mood distress (POMS > 60) and moderate depression (BDI > 20). Psychological burden was not influenced by current drug use, antiretroviral treatment, or viral load. At the 12-month evaluation, participants who received selenium reported increased vigor (p = 0.004) and had less anxiety (State, p = 0.05 and Trait, p = 0.02), compared to the placebo-treated individuals. No apparent selenium-related affect on depression or distress was observed. The risk for state anxiety was almost four times higher, and nearly nine times greater for trait anxiety in the placebo treated group, controlling for antiretroviral therapy, CD4 cell decline (> 50 cells) and years of education. CONCLUSIONS: Selenium therapy may be a beneficial treatment to decrease anxiety in HIV+ drug users who exhibit a high prevalence of psychological burden. PMID- 12906344 TI - Religious practice and depression among geriatric home care patients. AB - OBJECTIVE: To examine the relationship between religious practice and depression in a sample of geriatric patients receiving homecare nursing services. METHODS: Patients were sampled weekly for six months from all those aged 65 to 102, and newly enrolled in a visiting nurse agency (N = 130). Depression was assessed by home interviews using the SCID and HRSD. Patients reported their religious service participation prior to receiving homecare and currently. Health status, disability, pain, social support and history of depression were also assessed. RESULTS: The current prevalence of DSM-IV Major Depressive Disorder (MDD) was significantly greater (p < .05), and depressive symptoms were more severe (p < .02), among those persons who had not attended religious services prior to receiving homecare. Logistic regression demonstrated that the effect of religious attendance remained significant when controlling for health status, disability, pain, social support and history of depression. A subsequent analysis compared three groups of patients. They were those who had: 1) Not attended religious services; 2) Stopped attending since homecare; 3) Continued attending. Data demonstrated significantly decreasing prevalence of MDD (p < .03) across the groups. CONCLUSIONS: Prevalence of DSM-IV Major Depressive Disorder and the severity of depressive symptoms were significantly lower among homecare patients who attend religious services. Because a large proportion of persons stop attending religious services after initiating homecare, it is suggested that visitation by clergy may improve depressive symptoms for these patients. PMID- 12906345 TI - Quetiapine in the successful treatment of schizophrenia with comorbid alcohol and drug dependence: a case report. AB - BACKGROUND: Excluding nicotine and caffeine dependence, almost 50% of individuals with schizophrenia also meet the criteria for substance abuse or dependence. Comorbid drug abuse presents complications to the effective treatment of these patients because they have increased psychotic symptoms and poorer treatment compliance. CASE REPORT: This report describes thecase of a young man with schizophrenia and comorbid alcohol and cocaine abuse who was successfully treated with quetiapine. The patient was previously treated with olanzapine and developed priapism, which required emergency medical treatment. CONCLUSIONS: The possible utility of atypical antipsychotics in the treatment of patients with schizophrenia and comorbid substance abuse needs to be confirmed in clinical trials. PMID- 12906346 TI - Ammonia induced encephalopathy from valproic acid in a bipolar patient: case report. AB - Valproic acid is widely used as a mood stabilizer. We report a case of an adult with bipolar disorder taking therapeutic doses of valproic acid, who presented to the emergency department with coma related to hyperammonemia as a complication of valproic acid treatment. Valproic acid was discontinued which resulted in rapid clinical recovery. Valproic acid induced coma was likely related to a urea cycle enzymopathy. Clinicians should consider hyperammonemia in all patients who present with coma and other mental status changes while on valproic acid. In such patients, ammonia level should be obtained in addition to liver function tests. PMID- 12906347 TI - Drug addiction in sickle cell disease: case report. AB - There has been widespread speculation that patients with sickle cell disease (SCD) may become drug dependent if their painful crisis is treated with narcotics. However, there has been no scientific evidence to support this assertion. Paradoxically, individuals suffering from sickle cell disease who are not adequately treated may develop an addiction to narcotics due to self medication to treat their pain. In this article, we describe a 38-year-old African American woman who became addicted to cocaine due to self-medication of her sickle cell pain with cocaine. PMID- 12906348 TI - Sertraline-induced hyperglycemia: case report. AB - Empirical studies with humans suggest that selective serotonin reuptake inhibitors (SSRIs) may, through increases in serotonergic activity, increase insulin secretion, increase insulin sensitivity, and lower serum glucose levels. Indeed, the potentially beneficial effects of SSRI treatment in diabetics are documented. However, we describe the case of a female, with diet-controlled, type 2 diabetes, whose glucose levels increased with exposure to sertraline. PMID- 12906349 TI - Cognitive, behavioral, and psychiatric symptoms in two children with agenesis of the corpus callosum: case report. AB - OBJECTIVE: To report two children, with normal intelligence, referred for evaluation of complex cognitive, behavioral, and psychiatric problems, in which Agenesis of Corpus Callosum (ACC) was an incidental finding. METHOD: Case descriptions are used and a comprehensive differential diagnosis made with previous diagnoses of partial complex seizures (PCS), psychosis, anxiety, attention deficit hyperactivity disorder (ADHD), and behavior disorder. RESULTS: Due to the presence of multiple biopsychosocial interactions a multimodal intervention including pharmacological, behavioral, psychotherapeutic, and social approaches was implemented, with good results. CONCLUSION: These cases underscore the importance of conducting a comprehensive neuropsychiatric evaluation in children with severe behavior problems and other confusing symptoms. Since the incidence of ACC in behaviorally disturbed children, with normal intelligence, is unknown, physicians must accept that a reasonable degree of suspicion is warranted. Further studies are needed to facilitate physicians' awareness. PMID- 12906350 TI - Nutritional characteristics of the leaves of native plants growing in adverse soils of humid tropical lowlands. AB - Acid sulfate, peat, sandy podzolic, and saline soils are widely distributed in the lowlands of Thailand and Malaysia. The nutrient concentrations in the leaves of plants grown in these type of soils were studied with the aim of developing a nutritional strategy for adapting to such problem soils. In sago and oil palms that were well-adapted to peat soil, the N, P, and K concentrations were the same in the mature leaves, while the Ca, Mg, Na, and Fe concentrations were higher in the mature leaves of the oil palm than of the sago palm. Melastoma malabathricum and Melaleuca cajuputi plants that were well-adapted to low pH soils, peat. and acid sulfate soils were also studied. It was observed that a high amount of Al accumulated in the M. marabathricum leaves, while Al did not accumulate in M. cajuputi leaves. M. cajuputi plants accumulated large amounts of Na in their leaves or stems regardless of the exchangeable Na concentration in the soil, while M. malabathricum that was growing in saline-affected soils excluded Na. Positive relationships between macronutrients were recognized between P and N, between K and N, and between P and K. Al showed antagonistic relationships with P, K, Ca, Mg, Fe, Zn, Cu, and Na. Na also showed antagonistic relationships with P, K, Zn, Mn, Cu, and Al. Fe showed weak antagonistic relationships with Zn, Mn, Cu, and Al. PMID- 12906351 TI - Mineral characteristics of leaves of plants from different phylogeny grown in various soil types in the temperate region. AB - The objective of this research was to analyze selected minerals in leaves of plants, belonging to 166 species growing in alluvial, low pH, brown forest and serpentine soils. Mineral characteristics of the soils involved were also determined. For the macronutrients, in trees grown in alluvial soil, N, P, Ca, and Mg concentrations of leaves were higher in recently evolved plants than in plants with a longer period of evolution; K concentration remained constant regardless of evolution. In grasses grown in alluvial soil, it was difficult to detect the general tendency of mineral concentration. N, P, and K concentrations in alluvial soil were closely related to those in low pH and serpentine soils. Ca concentration in alluvial soil was lower than that in low pH and serpentine soils. Mg concentration in alluvial soils was higher than that in low pH soils, while lower than that in serpentine soil. Therefore, N, P, and K accumulated according to the plant characteristics for these elements, while Ca and Mg accumulation was strongly affected by the soil properties. For the micronutrients, in trees, Fe and Mn remained constant regardless of evolution; Zn concentration was lower in recently-evolved plants than in plants with a longer period of evolution. In grasses, Fe, Mn, and Zn concentrations in Caryophyllids were high. Except for Caryophyllids, Fe and Cu concentrations remained constant. Mn concentration decreased with evolution, Zn concentration was higher in recently-evolved plants than in plants with a longer period of evolution. PMID- 12906352 TI - Mineral characteristics of the leaves of 166 plant species with different phylogeny in the temperate region. AB - The P-N regression coefficient was moderate in Magnoliales, Coniferopsidae, Pteridophyta, and Asterids, but very high in Caryophyllids, and very low in Rosids. The K-N regression coefficient in trees remained constant regardless of evolution, and that in grasses was high except for Rosids compared with that in trees. The coefficient was very high in Caryophyllids and was very low in Rosids. The N-Ca and N-Mg relationship was not estimated at all, suggesting that the mechanism of Ca and Mg accumulation was completely different from that of N related accumulation. The Zn and Cu concentrations were related to the N concentration. The Al concentration in leaves was negatively correlated with the N, P, K, Ca, Mg, Na, Cu, Zn, Mn, and Fe concentrations, while the N, P, K, and Mn concentrations in leaves increased slightly with the increase of Al concentration in the high Al concentration. The Na concentration in leaves related negatively to the P, Ca, Cu, Zn, Mn and Al concentration range. Thus, it was demonstrated that most of the minerals in leaves accumulate negatively Al and Na indicating that there are antagonistic mechanisms for mineral accumulation in leaves among Al or Na and other mineral elements. PMID- 12906353 TI - Inactivation of trypsin inhibitors in sweet potato and taro tubers during processing. AB - In order to understand the extent of elimination of trypsin inhibitors during processing of sweet potato (Ipomoea batatas) and taro (Colocasia esculenta) tubers, a detailed study was conducted using tubers processed by oven drying, cooking, and microwave baking. Between 80 and 90% trypsin inhibitor (TI) activity was retained in sweet potato chips up to 2h at 70 degrees C. Among the four cultivars of sweet potatoes, RS-III-2 trypsin inhibitors were more heat labile. Heating at 100 degrees C led to rapid inactivation of TI of sweet potatoes. Varietal differences in thermal stability were more pronounced for the trypsin inhibitors of taro than sweet potatoes. Taro inhibitors were also more rapidly inactivated than sweet potato TI. Between 17 and 31% TI activity was retained in cooked tuber pieces of sweet potatoes, while only 3-10% were retained in taro cultivars. Very effective inactivation of trypsin inhibitors of sweet potatoes and taro could be obtained through microwave baking. Flour prepared from taro was devoid of TI activity, while 5-12% TI activity was retained in the flour prepared from sweet potatoes. The study clearly established that among the four techniques used, microwave baking and flour preparation were the best methods to eliminate TI from sweet potatoes and taro. PMID- 12906354 TI - Sensory and physical properties of peanut brittle prepared in different types of cookware and with different levels of sodium bicarbonate. AB - Two studies to evaluate peanut brittle containing either raw or roasted peanuts were conducted. In the first study, brittles were prepared in iron, aluminum, stainless steel and nonstick stainless steel pans. In the second study, brittles containing 0, 2 or 4 g of sodium bicarbonate prepared in either stainless steel or nonstick stainless steel pans were evaluated. All brittles were evaluated for color, shear strength and sensory properties at 0, 30 and 60 days. Products prepared in a nonstick stainless steel pan and those with added sodium bicarbonate were lighter, more yellow and slightly greener than other products. Pan type did not affect shear force but increasing sodium bicarbonate was associated with decreasing force needed to shear brittles. Peanut and caramel aromas were not affected by pan type or sodium bicarbonate level but were greater (p < 0.05) in products containing roasted peanuts. Burned flavors were more prevalent in products prepared in iron or stainless steel pans with roasted peanuts. More intense sweet and buttery flavors were detected as level of sodium bicarbonate increased. PMID- 12906355 TI - Physical behavior of purified and crude wax obtained from sunflower (Helianthus annuus) seed oil refineries and seed hulls. AB - The sunflower seed waxes obtained from two sources (i) seed hull as a standard and (ii) crude wax from oil refineries were studied for their crystallization, melting characteristics and morphology of crystals. The results of differential scanning calorimetry of wax obtained from seed hulls showed the melting temperature range of 13.18 degrees C with the onset at 62.32 degrees C, for purified wax, compared to the melting range of 24.73 degrees C with the onset at 42.3 degrees C. for crude wax. The enthalpy of fusion for both waxes were 57.55 mcal/mg and 7.63 mcal/mg, respectively. The DSC melt crystallization temperature range was 15.79 degrees C with the onset of 64.58 degrees C for purified wax and temperature range of 31.45 degrees C with an onset of 57.76 degrees C for crude wax. A similar pattern was observed of wax obtained from the crude wax of oil refineries. The enthalpy of crystallization was -64.27 mcal/mg and -7.67 mcal/mg, respectively. The purified wax obtained from the two sources (i) and (ii) were comparable with completion temperatures of 75.5 degrees C and 75.1 degrees C, respectively. The effect of inhibitor (lecithin) on crystallization of purified wax under light microscope and surface structure by scanning electron microscope were observed. Lecithin at 0.2% inhibited the crystallization but nucleation was unaltered. The wax crystal was inhibited to around 60% of the original size with 0.2% lecithin. It is concluded that the sunflower waxes studied were not comparable in their crystal properties of crude and purified states. Lecithin inhibited the crystallization of sunflower seed wax. PMID- 12906356 TI - Nonomuraea terrinata sp. nov., a novel soil actinomycete. AB - The taxonomic position of a soil isolate, strain E626, was evaluated using the polyphasic approach. The organism was found to have chemical and morphological features consistent with its assignment to the genus Nonomuraea, a member of the family Streptosporangiaceae. Strain E626 consistently formed a distinct phyletic line within the Streptosporangiaceae 16S rDNA tree using four different algorithms. Furthermore, the taxonomic distinctness of the organism is underpinned by a range of phenotypic properties, notably morphological features. It is, therefore, proposed that the organism be classified in the genus Nonomuraea as Nonomuraea terrinata sp. nov. PMID- 12906357 TI - Distribution and evolution of chitinase genes in Streptomyces species: involvement of gene-duplication and domain-deletion. AB - Streptomyces coelicolor A3(2) possesses nine genes for family 18 chitinases and two for family 19, showing high multiplicity. By hybridization analyses, distribution of those chitinase genes was investigated in six other Streptomyces species covering the whole phylogenetic range based on 16S rDNA sequences. All strains showed high-multiplicity of chitinase genes, like S. coelicolor A3(2). The phylogeny and gene organization of the family 18 chitinase genes cloned from Streptomyces species so far were then analyzed to investigate the gene evolution. It was concluded that Streptomyces already possessed a variety of chitinase genes prior to branching into many species, and that the ancestral genes of chiA and chiB have been generated by gene-duplication. In the course of the analyses, evidence that the chi30 and chi40 genes of S. thermoviolaceus were derived from their corresponding original chitinase genes by losing gene parts for substrate binding domains and fibronectin type III-like domains was obtained. It was thus shown that gene-duplication and domain-deletion were implicated in generating the high diversity and multiplicity of chitinase genes in Streptomyces species. PMID- 12906358 TI - Design of catabolic cassettes for styrene biodegradation. AB - A broad-host range metabolic cassette has been designed that, under the control of the Ptac promoter, expresses the sytABCD catabolic genes from Pseudomonas sp. Y2, which are responsible for the transformation of styrene into phenylacetic acid (styrene upper pathway). This novel cassette confers to phenylacetic acid degrading bacteria the ability to grow efficiently on styrene as the sole carbon and energy source. By combining both the sty cassette and the archetypal pWW0 TOL plasmid into the well-known Pseudomonas putida F1 aromatic biodegrader, we have constructed a novel derivative strain that shows one of the largest degradative potentials so far described for aromatic hydrocarbons, because it is able to use BTEX compounds (benzene, toluene, ethylbenzene and xylenes) and styrene as a source of carbon and energy. Furthermore, the sty cassette was engineered within a mini-transposon and endowed with a gene containment system, based on the toxic effect of the colicin E3 RNase, to reduce its lateral spread to other hosts. This contained cassette lacks defined transcriptional regulatory signals and, thus, it becomes an alternative strategy to select recombinant strains that efficiently express the desired phenotype from housekeeping regulatory elements. PMID- 12906359 TI - Implication of two glutathione S-transferases in the optimal metabolism of m toluate by Sphingomonas yanoikuyae B1. AB - A putative glutathione S-transferase (GST) gene (bphK) was identified in the meta cleavage operon for the degradation of m-toluate by Sphingomonas yanoikuyae B1. Disruption of bphK resulted in the loss of GST activity against 1-chloro-2,4 dinitrobenzene and a much increased lag time of the mutant strain MB3 (bphK::Km) following subculture into m-toluate medium. In contrast, an increased lag time was not observed when MB3 was grown on biphenyl or m-xylene and MB3 showed normal growth on m-toluate when complemented with a subclone containing the bphK gene only. Furthermore, an additional GST activity was detected in MB3. The induction timing of this second GST activity coincided with the beginning of the exponential growth phase of MB3 on m-toluate, reached maximal activity within three hours, and then dropped sharply to the basal level. Thus, it is apparent that BphK and/or the second GST are necessary for optimal growth of B1 on m toluate. PMID- 12906360 TI - Glyceraldehyde-3-phosphate dehydrogenase, a glycolytic enzyme present in the periplasm of Aeromonas hydrophila. AB - This is the first report describing the glycolytic enzyme, glyceraldehyde-3 phosphate dehydrogenase (GAPDH), as a protein associated with the cell envelope of a gram-negative bacterium (Aeromonas hydrophila). Dose-dependent GAPDH activity was detected in whole bacterial cells from exponentially growing cultures, indicating that an active form of GAPDH is located outside the plasma membrane. This activity represents roughly 10-20% of total cell activity, and it is not reduced by pretreatment of the cells with trypsin. Assays with soluble GAPDH indicate that the activity measured in intact cells does not originate by rebinding to intact cells of cytosolic enzyme released following cell lysis. GAPDH activity levels detected in intact cells varied during the growth phase. The relationship between GAPDH activity and cell culture density was not linear, showing this activity as a major peak in the late-logarithmic phase (A600 = 1.1 1.3), and a decrease when cells entered the stationary phase. The late exponential growing cells showed a GAPDH activity 3 to 4-fold higher than early growing or stationary cells. No activity was detected in culture supernatants. Enzymatic and Western-immunoblotting analysis of subcellular fractions (cytosol, whole and outer membranes, and periplasm) showed that GAPDH is located in the cytosol, as expected, and also in the periplasm. These results place the periplasmic GAPDH of A. hydrophila into the family of multifunctional microbial cell wall-associated GAPDHs which retain their catalytic activity. PMID- 12906361 TI - Numerical phenetic classification of clinically significant aerobic sporoactinomycetes and related organisms. AB - Clinically significant aerobic sporoactinomycetes, notably agents of mycetoma, were examined for a balanced set of unit characters and the resultant data analysed using standard numerical taxonomic procedures. All save two of the one hundred and seventy three tested strains were assigned to three multimembered cluster-groups, which encompassed sixteen major (4-7 strains), ten minor (2-3 strains) and forty single membered clusters, in an analysis based on the simple matching coefficient and unweighted pair group method with arithmetic averages algorithm. The three cluster-groups were equated with the genus Actinomadura (including Actinocorallia and Streptomyces somaliensis strains), and the genera Nocardiopsis and Streptomyces, and Thermobifida and Thermomonospora, respectively. In a corresponding principal co-ordinates analysis four multimembered groups corresponding to the genera Actinomadura, Nocardiopsis, Streptomyces, and Thermobifida and Thermomonospora were recognised. The causal agents of actinomycetoma were not only assigned to established taxa, notably, to Actinomadura latina, Actinomadura madurae, Actinomadura pelletieri and Streptomyces somaliensis, but also to additional centres of taxonomic variation which were equated with the rank of species. Most of the streptomycetes isolated from clinical material were assigned to clusters equated with the species Streptomyces albus and Streptomyces anulatus. The numerical taxonomic data were used to generate a frequency matrix designed to facilitate the identification of clinically significant Actinomadura, Nocardiopsis and Streptomyces strains to the species level; rapid enzyme tests accounted for eleven out of the twenty-one diagnostic tests. PMID- 12906362 TI - Transporters involved in uptake of di- and tricarboxylates in Bacillus subtilis. AB - Di- and tricarboxylates found as intermediates in the tricarboxylic acid cycle can be utilized by many bacteria and serve as carbon and energy source under aerobic and anaerobic conditions. A prerequisite for metabolism is that the carboxylates are transported into the cells across the cytoplasmic membrane. Bacillus subtilis is able to metabolize many di- and tricarboxylates and in this overview the available data on all known and putative di- and tricarboxylate transporters in B. subtilis is summarized. The B. subtilis transporters, that are of the secondary type, are discussed in the context of the protein families to which they belong. Available data on biochemical characterization, regulation of gene expression and the physiological function is summarized. It is concluded that in B. subtilis multiple transporters are present for tricarboxylic acid cycle intermediates. PMID- 12906363 TI - Pheochromocytoma: a permanent challenge for the clinical endocrinologist. PMID- 12906364 TI - Urinary iodine concentration during pregnancy in an area of unstable dietary iodine intake in Switzerland. AB - We prospectively investigated urinary iodine concentration (UIC) in pregnant women and in female, non-pregnant controls in the canton of Berne, Switzerland, in 1992. Mean UIC of pregnant women [205 +/- 151 microg iodine/g creatinine (microg l/g Cr); no. = 153] steadily decreased from the first (236 +/- 180 microg l/g Cr; no. = 31) to the third trimester (183 +/- 111 microg l/g Cr, p < 0.0001; no. = 66) and differed significantly from that of the control group (91 +/- 37 microg l/g Cr, p < 0.0001; no. = 119). UIC increased 2.6-fold from levels indicating mild iodine deficiency in controls to the first trimester, demonstrating that high UIC during early gestation does not necessarily reflect a sufficient iodine supply to the overall population. Pregnancy is accompanied by important alterations in the regulation of thyroid function and iodine metabolism. Increased renal iodine clearance during pregnancy may explain increased UIC during early gestation, whereas increased thyroidal iodine clearance as well as the iodine shift from the maternal circulation to the growing fetal-placental unit, which both tend to lower the circulating serum levels of inorganic iodide, probably are the causes of the continuous decrease of UIC over the course of pregnancy. Mean UIC in our control group, as well as in one parallel and several consecutive investigations in the same region in the 1990s, was found to be below the actually recommended threshold, indicating a new tendency towards mild to moderate iodine deficiency. As salt is the main source of dietary iodine in Switzerland, its iodine concentration was therefore increased nationwide in 1998 for the fourth time, following increases in 1922, 1965 and 1980. PMID- 12906365 TI - The effect of surgical stress on insulin sensitivity, glucose effectiveness and acute insulin response to glucose load. AB - Hyperglycemia after stress is a very common clinical phenomenon. It is generally hypothesized that the underlying cause is a neuroendocrine-mediated deterioration in glucose metabolism. However, the detailed roles of insulin sensitivity, glucose effectiveness and acute insulin response to glucose load in response to stress have not been well established. Hernioplasty was used as a minor stress model for studying stress-induced hyperglycemia. Eleven healthy young men were enrolled voluntarily in this study. Their mean age was 22.0 +/- 0.9 yr and BMI 23.3 +/- 0.6 kg/m2. Frequently sampled i.v. glucose tolerance tests were performed one day before and one day after the surgery. Insulin sensitivity (SI), glucose effectiveness (EG) and area under acute insulin response (AIR) were calculated from "minimal model" algorithms. We also measured fasting concentrations of human GH, ACTH and F on the days of the test. Compared to the pre-operation data, levels of ACTH and F did not change significantly after the surgery. Only GH levels were marginally significant. On the other hand, the SI (0.75 +/- 0.1, 0.52 +/- 0.9 x 10(-5) min(-1)/pmol, p = 0.04), EG (0.023 +/- 0.03, 0.016 +/- 0.003 min(-1), p = 0.01) and AIR (6738.5 +/- 1111.6, 5130.0 +/- 1047.2 pmol, p = 0.005) were all significantly decreased after surgery. The percentages of decrease were 16.3 +/- 15.5, 32.1 +/- 10.3 and 17.8 +/- 10.3%, respectively. Finally, only the changes of EG positively correlate with the changes of ACTH before and after surgery. No significant changes were noted among other stress hormones and the changes of SI, EG and AIR. In conclusion, hernioplasty results in reduced SI, EG and AIR. Among them, although not statistically significant, the EG showed the most distinct decrease after the surgery, which has not been found in previous literature. PMID- 12906366 TI - Caffeine raises the serum melatonin level in healthy subjects: an indication of melatonin metabolism by cytochrome P450(CYP)1A2. AB - Caffeine is metabolized in the liver by cytochrome P450(CYP)1A2. Recent findings imply that this enzyme may also be of importance for the metabolism of human melatonin (MT). If caffeine and MT are metabolized by the same enzyme, one may expect to find different serum MT levels after ingestion of coffee compared with placebo. Although coffee is consumed by people all over the world, few studies have focused on whether caffeine actually affects serum MT levels in normal subjects. We decided to study that particular topic. For that purpose 12 healthy individuals were tested on two occasions, one week apart. On one of these occasions they were given a capsule containing 200 mg caffeine in the evening. On the other, they received placebo. The experimental order was randomized. Serum MT levels were determined every second hour between 22:00 h and 08:00 h, and the melatonin areas under the curve (MT-AUCs) were calculated. After caffeine the serum MT level rose from 0.09 +/- 0.03 nmol/l at 22:00 h to 0.48 +/- 0.07 nmol/l at 04:00 h. The corresponding rise after placebo was less prominent (from 0.06 +/ 0.01 to 0.35 +/- 0.06 nmol/l). This was reflected by the MT-AUC which was 32% larger after ingestion of caffeine compared with placebo (MT-AUC(caffeine) 3.16 +/- 0.44 nmol/l x h vs MT-AUC(placebo) 2.39 +/- 0.40 nmol/l x h; p < 0.02). These findings imply that caffeine, ingested in the evening at a dose corresponding to two ordinary cups of coffee, augments the nocturnal serum MT level, which in turn supports the notion that cytochrome P450(CYP)1A2 is involved in the hepatic metabolism of human MT. PMID- 12906367 TI - Hypothalamo-pituitary-adrenal axis in acute myocardial infarction treated by percutaneous transluminal coronary angioplasty: effect of time of presentation. AB - Acute myocardial infarction (AMI) is associated with a stimulation of cortisol which lasts 24 hours in patients treated by thrombolysis. Percutaneous transluminal coronary angioplasty (PTCA) is an alternative treatment for AMI which reduces the length of myocardial ischemia. Our objective was the determination of the amplitude and duration of cortisol and other hormones of the hypothalamo-pituitary-adrenal (HPA) axis release in patients undergoing PTCA. These responses were also analyzed in relation with the time of onset of AMI. The effect of coronarography with or without angioplasty in patients without AMI was also studied. Plasma ACTH, cortisol, corticotropin-releasing hormone and arginine vasopressin levels were determined during the first 48 hours in 20 patients with first AMI, treated by PTCA and in 10 patients without AMI undergoing coronarography (and angioplasty in five of them). A strong stimulation of the HPA axis was observed in AMI patients, but the duration of cortisol secretion was significantly reduced (less than 8 hours) as compared with previous studies in patients treated with thrombolysis. A clear-cut ACTH-cortisol dissociation was also observed after the third hour. ACTH and cortisol stimulation was higher in patients admitted between 04:00 h and 16:00 h than in patients admitted between 16:00 h and 04:00 h In patients without AMI, coronarography induced a moderate, but significant short-lasting ACTH and cortisol stimulation. In conclusion, our data suggest that the degree of stimulation of the HPA axis may depend upon the type of treatment and the circadian rhythm of this axis. PMID- 12906368 TI - Lipoprotein (a) in childhood: correlations with family history of cardiovascular disease. AB - The association between lipoprotein (a) [Lp(a)] and cardiovascular diseases is well known. Lp(a) is an independent risk factor for the development of atherosclerosis. Little information concerning Lp(a) during childhood is available. The aim of the present investigation was to determine the Lp(a) concentration in a cohort of children aged between 4 and 15 yr and to correlate Lp(a) with: a) overweight status; b) body fat distribution; c) family history of vascular diseases in their parents and grandparents. Six hundred and eighty-nine children (350 males, 339 females), were enrolled in the study. BMI as index of being overweight was calculated; the waist-to-hip ratio (WHR) and the waist-to thigh ratio (WTR) were calculated to obtain two anthropometric indexes for the pattern of body fat distribution. The areas of visceral (VAT) and subcutaneous adipose tissue (SAT) were evaluated by MRI at the L4-L5 level in only 30 overweight subjects. The serum of Lp(a), total cholesterol (TC), HDL-cholesterol, LDL-cholesterol and triglycerides were evaluated in the whole population. Moreover, the same biochemical study was performed in 70 children's parents randomly chosen. A structured questionnaire was administered to the children's parents to investigate the presence of cardiovascular disease (CVD) in family stories. Our data show no Lp(a) serum differences between children according to sex, age and body composition. The strong correlation between the children's and the parents' Lp(a) concentrations and the occurrence of CVD in their grandparents suggests that Lp(a) represents an important independent early risk factor for the development of CVD in adulthood. Measurements of Lp(a) in childhood may help to evaluate this risk in subjects with family history of cardiovascular diseases. PMID- 12906370 TI - Lesion of the amygdala on the right and left side suppresses testosterone secretion but only left-sided intervention decreases serum luteinizing hormone level. AB - The effect of right- and left-sided intra-amygdaloid injection of kainic acid on the hypothalamo-hypophyseal-testicular axis was studied in rats. Both right- and left-sided injection of the neurotoxin into the amygdala resulted in a significant decrease in basal testosterone secretion in vitro of both testes and in serum testosterone concentration. In addition, left-sided administration of kainic acid significantly suppressed serum luteinizing hormone level, while right sided intervention did not alter this parameter. The results of the present study provide further evidence on the involvement of the amygdala in the control of testicular steroidogenesis. Furthermore, the observations suggest functional asymmetry of the amygdala concerning the mechanism of suppressed testosterone secretion. PMID- 12906369 TI - Three weekly injections (TWI) of low-dose growth hormone (GH) restore low normal circulating IGF-I concentrations and reverse cardiac abnormalities associated with adult onset GH deficiency (GHD). AB - GH replacement therapy given 3 times weekly (TWI) and adjusted to allow serum IGF I concentrations in the mid-normal range for sex and age has been shown to be as effective as the daily regimen in improving lipid profile, body composition, bone mass and turnover in adult GH deficient (GHD) patients. Only one study has investigated so far the short-term (6 months) effect of a fixed weight-based TWI dosing schedule on heart structure and function in childhood onset (CO) GHD patients, whereas such a schedule in adult onset (AO) GHD patients has not been studied as yet. Aim of this study was to investigate whether a 1-yr low-dose titrated TWI GH-replacement regimen aimed at achieving and maintaining IGF-I levels within the low normal limits for age and sex is able to affect cardiovascular and heart parameters in a group of AO GHD patients. Eight adult patients (4 women and 4 men, age 35.8 +/- 3.37 yr, body mass index, BMI, 28.7 +/- 2.62 kg/m2) with AO GHD were included in the study, along with 10 healthy subjects, matched for age, sex, BMI and physical activity (6 women and 4 men, age 35.2 +/- 4.05 yr, BMI 28.4 +/- 2.34 kg/m2). M- and B- mode ecocardiography and pulsed doppler examination of transmitral flow were performed in GHD patients at baseline and after 3 and 12 months of GH therapy (mean GH dose 6.7 +/- 0.8 microg/kg/day given thrice a week), while normal subjects were studied once. Treatment with GH for 1 yr induced a significant increase in left ventricular (LV) diastolic and systolic volumes (+11.1 and +16.5%, respectively). Systolic LV posterior wall thickness and LV mass were increased (+10.2 and +7.7%, respectively) by GH administration. Systemic vascular resistance was significantly decreased by 1-yr GH therapy (-13.8% after 1 yr), while stroke volume, cardiac output and cardiac index were increased (+9.4, +11.6 and + 11.9%, respectively). LV end-systolic stress was decreased at the end of GH therapy ( 11.2%). E and A wave, significantly reduced at baseline, were increased by 1 yr of GH therapy (+23.3% and +28.1%, respectively); likewise, the abnormally high E peak deceleration time was partially reversed by GH administration (-10.7%). Our study, though conducted in a small sample size, demonstrates that a TWI GH treatment schedule is able to reverse the cardiovascular abnormalities in AO GHD patients and to improve body composition and lipid profile. The maintenance of circulating IGF-I concentrations within the low normal range allows to avoid most of the side-effects reported with higher GH doses while being cost-effective and improving the patient's compliance. PMID- 12906372 TI - Riedel's thyroiditis and fibrous variant of Hashimoto's thyroiditis: a clinicopathological and immunohistochemical study. AB - The aim of this study was to analyze and compare clinico pathological aspects of Riedel's thyroiditis (RT) and the fibrous variant of Hashimoto's thyroiditis (HTFV), and to show their immunohistochemical features. We reviewed 6 cases of HTFV and 4 cases of RT. Compared to RT, HTFV patients had hypothyroidism, no pressure symptoms, and frequently diagnostic fine-needle aspiration biopsy (FNAB) cytology. At histology, invasion of surrounding tissues and presence of occlusive phlebitis distinguished RT from HTFV. At immunohistochemistry, RT--compared to HTVF--was characterized by: 1) a more abundant fibrous reaction, and granulocytic, monocytic and eosinophil infiltration; 2) few plasma-cells, CD8+ T- and B-lymphocytes. The results of our study add further evidence regarding the separation of RT and HTFV in their peculiar clinical, laboratory, cyto histological and immunohistochemical aspects. PMID- 12906371 TI - Effect of thyroid hormone on the development and gene expression of hormone receptors in rat testes in vivo. AB - Thyroid hormone is known to play a pivotal role in the regulation of prepuberal rat testes development and function with specific influence on the differentiation of Sertoli cells, the only cell type that expresses thyroid hormone receptors in testes. To explore in vivo effects of thyroid hormone on testes development and the regulation of testicular gene expression, the hyper- and hypothyroid rat models were established by T3 injection to pups (ip 100 microg/kg bw) and by oral administration of 6-N-propyl-2-thiouracil (PTU) to the lactating mother from days 1 to 21 post-delivery. Half of the rats from each group were sacrificed at 21 days of age, and the other half were allowed to recover with discontinued treatments from day 22 to day 50. At 21 days of age, a significantly elevated serum T3 level was observed in hyperthyroid rats (179.5 ng/dl) vs controls (97.5 ng/dl), and in hypothyroid rats a significantly lower level of T3 was detected (26.1 ng/dl). However, serum T4 concentration was significantly lower in both hyper- (0.105 microg/dl) and hypothyroid (0.058 microg/dl) rats compared to the controls (2.48 microg/dl). In recovered rats in which the serum T3 and T4 were restored to normal, the serum T levels remained remarkably lower in both hyper- and hypothyroid rats. The significantly decreased body and testes weights observed in both hyper- and hypothyroid rats at 21 days of age were not restored by the time they were 50 days old. Histological analyses of testes of 21-day-old hypothyroid rats revealed smaller-sized seminiferous tubules, incomplete lumen formation and delayed germ cell differentiation and in hyperthyroid rats an increased number of early stage spermatocytes was found. Testicular mRNA levels of follicle-stimulating hormone receptor (FSH-R), luteinizing hormone receptor (LH-R) and androgen binding protein (ABP) were studied by Northern blot hybridization. At 21 days of age data showed that FSH-R mRNA levels were significantly higher in both hyper- and hypothyroid rat testes compared to controls, but no differences were detected in recovered 50-day-old rats. Significantly decreased ABP mRNA levels were detected only in hypothyroid rat testes compared to those in both the hyperthyroid and control groups at 21 days of age, but no significant change was observed in recovered 50-day-old rats. To further evaluate the effect of thyroid hormone on the Leydig cell function, the 2.3/2.6 kb specific LH-R hybridization bands were detected with rat LH-R cRNA probe. Significant suppression of LH-R mRNA levels was only observed in the hypothyroid rat testes at 50 days of age. The testicular thyroid hormone receptors (TRs) and the regulation of TR by thyroid hormone were investigated using semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) assays. Both TRalpha and TRbeta mRNAs were identified in the testes from 21- and/or 50-day-old rats. TRalpha mRNA levels were significantly increased in hypothyroid rat testes and were suppressed in hyperthyroid rats at 21 days of age and no changes of TRalpha mRNA were found in recovered animals. Our in vivo data strongly suggest that the thyroid hormone directly affects the development of prepuberal testes and the regulation of FSH-R and ABP gene expression in Sertoli cells, as well as the LH-R mRNA levels in Leydig cells, which may lead to further modulating the effect of gonadotropins on testes function. PMID- 12906373 TI - Case report of hypopituitarism with suspected syndrome of inappropriate VP secretion (SIADH) due to a large aneurysm of the internal carotid in the sellar region. AB - Hypopituitarism and hyponatremia, especially when severe, are infrequent findings particularly when the cause of hypopituitarism at presentation is unknown and untreated. Interestingly, hyponatremia is usually seen in elderly patients with hypopituitarism due to various causes. We present a case with unrecognized and untreated hypopituitarism due to a large aneurysm of the internal carotid artery in the sellar region causing the syndrome of inappropriate secretion of antidiuretic hormone (SIADH). PMID- 12906374 TI - Interleukin-6 producing pheochromocytoma presenting with acute inflammatory syndrome. AB - Pheochromocytomas are tumors able to produce catecholamines and a variety of biologically active neuropeptides. We report the case of a 36-yr-old female patient with pheochromocytoma exhibiting headache, intermittent fever, thrombocytosis, and marked inflammatory signs. Nonsteroidal anti-inflammatory drugs were ineffective in lowering the body temperature, while a corticosteroid agent obtained excellent results. IL-6 was found elevated (20 pg/ml); it fell to 4.5 pg/ml 3 weeks after the adrenalectomy, in parallel to normalization of other laboratory data. The interleukin-6 (IL-6) over-production can either be ascribed directly to the tumor (as confirmed by immunohistochemistry) or indirectly accounted for by tumoral production, as a consequence of the high levels of circulating norepinephrine. To our knowledge, our paper represents the 6th case report of IL-6 secreting pheochromocytoma associated with clinical markers of inflammatory response. PMID- 12906375 TI - Clinical utility of bone markers in the evaluation and follow-up of osteoporotic patients: why are the markers poorly accepted by clinicians? PMID- 12906376 TI - Biochemical markers of bone metabolism in the assessment of osteoporosis: useful or not? PMID- 12906377 TI - Subclinical thyroid disorders: the menace of the Trojan horse. AB - This review aims at summarizing some new data regarding the so-called subclinical thyroid disease and incorporate the knowledge in order to provide a basis for a more careful approach to these morbidities. SHyper represents a considerable risk factor for atrial fibrillation in the elderly and also for osteoporosis, which is a major cause of morbidity in postmenopausal women. SHypo is not an innocuous condition. It progresses to thyroid failure in patients with positive thyroid antibodies, it affects neurobehavior, it may influence cardiac contractility at exercise and it is associated with an impaired lipid profile that may provoke atherosclerosis. Therefore, both conditions should be screened more carefully in the community and treatment should be more often considered. PMID- 12906379 TI - Leptin is not present in infant formulas. PMID- 12906378 TI - Testosterone replacement therapy in male hypogonadism. AB - In human males 6-7 mg of testosterone are secreted by the testes in a circadian rhythm with a nocturnal rise in testosterone followed by a decline during the day. Testosterone is necessary to induce and maintain secondary sexual characteristics, lean muscle mass, bone density and for normal sexual behaviour and cognitive function in men. Replacement therapy has been shown to be beneficial in men with overt hypogonadism. Natural testosterone should be used and not modified molecules. Testosterone is currently available in oral, intramuscular, subcutaneous and transdermal preparations. Recent advances in testosterone replacement therapy include testosterone gels which provide flexibility in dosing and minimal skin irritation resulting in good compliance, and the development of longer acting intramuscular preparations which result in more stable testosterone levels with longer injection intervals. All patients receiving testosterone should be carefully monitored for changes in hematocrit, liver function, lipid parameters and prostate specific antigen (PSA).This article reviews the current experience with the use of various forms of testosterone for the treatment of male hypogonadism. PMID- 12906380 TI - Prospore membrane formation: how budding yeast gets shaped in meiosis. AB - During meiosis in Saccharomyces cerevisiae four daughter cells, called spores, are generated within the boundaries of the mother cell. This cell differentiation process requires de novo synthesis of prospore membranes (PSMs), which are the precursors of the spore plasma membranes. Assembly of these membranes is initiated at the spindle pole bodies (SPBs) during meiosis II. At this stage of the cell cycle, 4 SPBs are present. Two different meiosis-specific structures are known to be required for PSM formation. At the SPBs, specialized attachments, called the meiotic plaques, provide the required functionality necessary for the recruitment and assembly of the membranes. During subsequent membrane elongation, a second structure becomes important. This proteinaceous assembly forms a coat, called the leading edge protein coat (LEP coat), which covers the boundaries of the membranes. Assembly of the coat occurs at sites next to the SPBs, whereas its disassembly is concomitant to the closure of the membranes. This mini review discusses our current understanding of how the meiotic plaque and the LEP coat might function during biogenesis of the prospore membrane. PMID- 12906381 TI - Screening of antagonistic bacteria for biological control of nursery wilt of black pepper (Piper nigrum). AB - Bacterial antagonists of Phytophthora capsici were isolated from underground shoot portions of rooted cuttings of black pepper. Initially isolates were screened by dual culture on potato dextrose agar and carrot agar. Further, a screening was done on black pepper shoots for supression of lesion caused by the pathogen. Most of the antagonists showed varying levels of antagonism in the dual culture and the shoot assay. Isolate PN-026, showing the highest suppression of lesion development in the shoot assay was found to be the most efficient antagonist in reducing Phytophthora capsici induced nursery wilt of black pepper. This screening involving the host, pathogen, and the antagonist, performed on black pepper shoot (the planting material for this vegetatively propagated crop), could be used as a rapid and reliable method for the isolation of efficient bacterial antagonists of P. capsici. PMID- 12906382 TI - Biodegradation of methyl tertiary butyl ether (MTBE) by a bacterial enrichment consortia and its monoculture isolates. AB - Methyl tertiary butyl ether (MTBE), an important gasoline additive, is a recalcitrant compound posing serious environmental health problems. In this study, MTBE-degrading bacteria were enriched from five environmental samples. Enrichments from Stewart Lake sediments and an MTBE contaminated soil displayed the highest rate of MTBE removal; 29.6 and 27.8% respectively, in 28 days. A total of 12 bacterial monocultures isolated from enrichment cultures were screened for MTBE degradation in liquid cultures. In a nutrient-limited medium containing MTBE as the sole source of carbon and energy, the highest rate of MTBE elimination was achieved with IsoSL1, which degraded 30.6 and 50.2% in 14 and 28 days, respectively. In a nutrient-rich medium containing ethanol and yeast extract, the bacterium (Iso2A) substantially removed MTBE (20.3 and 28.1% removal in 14 and 28 days, respectively). Based upon analysis of the 16s rRNA gene sequence and data base comparison, IsoSL1 and Iso2A were identified as a Streptomyces sp. and Sphingomonas sp., respectively. The Streptomyces sp. is a new genera of bacteria degrading MTBE and could be useful for MTBE bioremediation. PMID- 12906383 TI - Characterisation of Lysobacter enzymogenes (Christensen and Cook 1978) strain 3.1T8, a powerful antagonist of fungal diseases of cucumber. AB - Isolate 3.1T8 of Lysobacter enzymogenes (Christensen and Cook 1978), originating from the rhizosphere of cucumber and shown to have the potential to control Pythium aphanidermatum, is described. The strain produces extracellular proteases and lipases and shows high levels of resistance against streptomycin, kanamycin and tetracycline, but not to chloramphenicol. It shows strong in vitro antibiosis against P. aphanidermatum and several other phytopathogenic fungi. In order to identify the isolate, a carbon substrate oxidation profile (Biolog) was generated, and fatty acid methyl ester (FAME) analysis was performed. Also, the 16S rRNA gene was cloned and sequenced. With Biolog and FAME analysis, no assignment to species level was possible, because the species was not in the respective databases. BLAST analysis of the obtained sequence, followed by phylogenetic analysis, using a number of related and unrelated sequences, showed that the isolate was most closely related to Lysobacter enzymogenes (Christensen and Cook 1978). PMID- 12906384 TI - Cork stoppers industry: defining appropriate mould colonization. AB - AIMS: The main aims of this work were the study of cork slabs moulds colonization and the evaluation of the moulds diversity during cork processing steps, in different cork stoppers factories. Simultaneously, it was envisaged to perform an evaluation of the air quality. METHODS AND RESULTS: Moulds were isolated and identified from cork slabs and cork samples in four cork stoppers factories. The identification was based on morphological characters and microscopic observation of the reproductive structures. Airborne spore dispersion was assessed using a two stage Andersen sampler. It was observed that Chrysonilia sitophila was always present on cork slabs during the maturing period, but mould diversity appeared to be associated to the different factory configurations and processing steps. CONCLUSIONS: Spatial separation of the different steps of the process, including physical separation of the maturation step, is essential to guarantee high air quality and appropriate cork slabs colonization, i.e. C. sitophila dominance. The sorting and cutting of the edges of cork slabs after boiling and before the maturing step is also recommended. SIGNIFICANCE AND IMPACT OF THE STUDY: This study is very important for the cork stopper industry as it gives clear indications on how to keep high quality manufacturing standards and how to avoid occupational health problems. PMID- 12906385 TI - Species pattern and genetic diversity of Trichoderma in a mid-European, primeval floodplain-forest. AB - We investigated the occurrence and genetic diversity of Trichoderma in the river Danube national park, a primeval, riparian forest area located south-east of Vienna (Austria) which represents one of the last cases of an original European river-floodplain landscape. Forty-six strains were isolated and identified at the species level by analysis of morphological characters, by sequence analysis of their internal transcribed spacer regions 1 and 2 (ITS 1 and 2) of the rDNA cluster and--in some cases--a fragment of the translation elongation factor 1alpha (tef1) gene, and RAPD-analysis. Twenty-one strains were positively identified as T. harzianum, thirteen as T. rossicum, four as T. cerinum, two as T. hamatum, and one each as T. atroviride and T. koningii: four strains yielded two different ITS1 and 2 as well as tef1 sequence types, which were not alignable with any known species. Our studies show that they represent two new taxa of Trichoderma. PMID- 12906386 TI - Typing of Salmonella enterica subsp. enterica serovars Choleraesuis, Typhimurium, Dublin and laboratory strains of Escherichia coli using subtracted restriction fingerprinting (SRF). AB - The aim of the present study was to evaluate the usefulness of a new typing technique called subtracted restriction fingerprinting (SRF) for bacterial strain and isolate discrimination. The technique was applied to isolates of Salmonella enterica subsp. enterica (S.) serovars Choleraesuis, Typhimurium, Dublin and to two laboratory strains of E. coli. SRF is based on the selective removal of excess fragments from a restriction digest using magnetic particles. Subsequently, the remaining subset of restriction fragments can easily be analyzed with a conventional agarose gel. Larger fragments are preferentially removed by SRF. This results in an even distribution of bands within each electrophoretic lane and significantly improves scoring. The high discriminatory index for (S.) Choleraesuis (D = 0.914) illustrated the suitability of SRF for genome typing. PMID- 12906387 TI - Comparison of cross inoculation potential of South African avocado and mango isolates of Colletotrichum gloeosporioides. AB - Three hundred and eighteen Colletotrichum gloeosporioides isolates from stem-end rot and anthracnose infected avocados as well as from stem-end rot/anthracnose and soft brown rot on mango, were compared using fruit inoculations. Isolates could be categorised according to lesion size and both avocado and mango isolates produced larger lesions when inoculated on their own hosts. Cross-inoculation potential of these isolates was also compared on strawberries, peppers, guavas, papayas and citrus. All isolates produced lesions on all hosts except citrus. Factors such as area of origin and symptom type from which original isolations were made, could not be correlated with lesion development on these hosts. PMID- 12906388 TI - Rhizosphere microflora of plants used for the phytoremediation of bitumen contaminated soil. AB - The microbial communities and their degradative potential in rhizospheres of alfalfa (Medicago sativa) and reed (Phragmites australis) and in unplanted soil in response to bitumen contamination of soil were studied in pot experiments. According to the results of fluorescence microscopy, over a period of 27 months, bitumen contamination of soil reduced the total number of microorganisms more significantly (by 75%) in unplanted than in rhizosphere soil (by 42% and 7% for reed and alfalfa, respectively) and had various effects on some important physiological groups of microorganisms such as actinomycetes as well as nitrogen fixing, nitrifying, denitrifying, ammonifying, phosphate-solubilizing, sulphur oxidizing, cellulolytic and hydrocarbon-degrading microorganisms. The changes in the physiological structure of the microbial community under bitumen contamination were found to hinge on not merely the presence of plants but also their type. It was noted that the rhizosphere microflora of alfalfa was less inhibited by hydrocarbon pollution and had a higher degradative potential than the rhizosphere microflora of reed. PMID- 12906389 TI - Solubilization of inorganic phosphate and plant growth promotion by cold tolerant mutants of Pseudomonasfluorescens. AB - A study for screening and selection of cold tolerant mutants of Pseudomonas fluorescens strains GRS1, PRS9 and ATCC13525 based on 'P' solubilization ability and subsequent effect on plant growth promotion under in vitro and in situ conditions was conducted. Of all the mutants tested, two were selected, as there was a 21-fold increase in CRPF, (GRS, mutant) and a 10-fold decrease in CRPF7 (PRS9 mutant) over their respective wild types. Under in vitro conditions at 10 degrees C, these cold tolerant mutants exhibited increased plant growth indicating their functionality at low temperature. Subsequently, greenhouse trials using soil-plant microcosms were conducted which revealed that CRPF, (high 'P' solubilizer) was a good rhizosphere colonizer showing a significant increase in root (30 and 20%) and shoot length (20 and 24%) of mung bean, both in sterilized and unsterilized soil, respectively. On the contrary, CRPF, (low 'P' solubilizer) did not stimulate plant growth. Furthermore, sand experiments indicated that tricalcium phosphate served as better phosphorus source for CRPF2 treated mung bean seeds. PMID- 12906390 TI - Partial purification and characterization of the bacteriocin produced by Lactobacillus acidophilus YIT 0154. AB - One strain of Lactobacillus acidophilus was found to produce a bacteriocin-like substance in the culture filtrate. The substance was produced in a growth associated manner, showed heat stability at neutral and acidic pH and exhibited antibacterial activity against various species of Lactobacillus including L. acidophilus itself. The molecular weight of the substance was in the range of 6.2 95 kDa. N-terminal amino acid sequence analysis suggests that the substance may belong to class IIb bacteriocin. PMID- 12906391 TI - Evaluation of arbitrarily primed PCR for typing of Desulfovibrio desulfuricans strains. AB - Arbitrarily primed polymerase chain reaction (AP-PCR) method was applied to the differentiation of 15 (soil and intestinal) Desulfovibrio desulfuricans strains. The primer M 13, which is a core sequence of phage M 13, was found to be appropriate for the differentiation of isolates of this species. The analysis revealed characteristic band patterns for all of the examined strains of which two soil strains (DV-7 and DV-8) showed identical DNA fingerprints. According to Jaccard's coefficient, the soil bacterial group as well as intestinal bacterial group formed two different clusters. Furthermore, the soil strains showed greater variability than the intestinal isolates. Based on the AP-PCR fingerprints D. desulfuricans strains were differentiated depending on their origin. This study demonstrates that the typing method AP-PCR can be useful in epidemiologic investigations as a rapid and valuable tool for differentiation of the strains of D. desulfuricans species. PMID- 12906392 TI - Screening of Bifidobacterium strains isolated from human faeces for antagonistic activities against potentially bacterial pathogens. AB - As probiotic bacteria, strains belonging to the genus Bifidobacterium colonise the gastro-intestinal tract of humans and animals at the time of birth, and they are found in young as well as in adult individuals in great numbers. Moreover, they can interact with the development of enteric infections by the production of antimicrobial metabolites. In this work 281 strains of bifidobacteria were anaerobically isolated from human faecal samples, supplied by volunteers of different ages (youngs, adults, elders), and preliminarly described by microscopic observation. All strains were screened by the fructose 6-phosphate phosphoketolase (F6PPK) test in order to confirm their classification within the genus Bifidobacterium. Selected strains were used to evaluate their antagonistic activities against Escherichia coli, Salmonella thyphimurium, Staphylococcus lentus, Enterococcus faecalis, Acinetobacter calcoaceticus, Sphingomonas paucimobilis, Listeria monocytogenes, Yersinia enterocolitica, Bacillus cereus, Clostridium sporogenes. Experiments were performed in vitro by different methods based on the observation of growth inhibition in Petri dishes. The strains that showed the highest inhibiting activities were compared by SDS-PAGE for total cell proteins, using type strains of human origin as references. Representative isolates were metabolically characterised by the BIOLOG system; a specific database was created with strains obtained from our collection and a statistical evaluation for metabolic patterns was carried out. PMID- 12906393 TI - Cell surface characteristics of two halotolerant strains of Sinorhizobium meliloti. AB - The halotolerant Sinorhizobium meliloti strain Rmd201 and its variant Rmd201 a were examined for their cell surface properties. The variant strain formed rough colonies and was found to be more hydrophobic. Growth of the variant strain was not affected appreciably when NaCl concentration of the medium was increased from 2 mM to 700 mM. Exopolysaccharide (EPS) and the lipopolysaccharide (LPS) content of the variant strain was found to be 7 and 14 times less, respectively, than the parent strain. However, enhanced synthesis of high molecular weight LPS bands were observed in SDS-PAGE analysis in the variant strain when the NaCl concentration was raised from 2 mM to 700 mM. Ribose and glucosamine were present in the variant LPS only. Mannose appeared as a major LPS constituent of the variant when grown in high salt containing medium. All these cell surface characteristics indicated that there were significant differences between the halotolerant strains of S. meliloti. The changes in the cell surface of the variant strain indicated the possible mutation in the gene(s) of cell surface polysaccharide biosynthesis. PMID- 12906394 TI - Effect of carbon source on pyrimidine biosynthesis in Pseudomonas alcaligenes ATCC 14909. AB - The effect of carbon source on the regulation of the de novo pyrimidine biosynthetic enzymes in Pseudomonas alcaligenes ATCC 14909 was investigated. The de novo pyrimidine biosynthetic enzymes were measured in extracts of P. alcaligenes ATCC 14909 cells and of cells from an auxotroph deficient for orotate phosphoribosyltransferase activity. Pyrimidine biosynthetic enzyme activities in ATCC 14909 were influenced by pyrimidine supplementation to the culture medium but not by the carbon source present. Pyrimidine limitation of the auxotroph elevated the de novo enzyme activities indicating that this pathway may be controlled at the transcriptional level by a pyrimidine-related compound. Its regulation seemed to be subject to less transcriptional control by a pyrimidine related compound than what was observed in the closely related species Pseudomonas pseudoalcaligenes. PMID- 12906395 TI - Experimental murine cryptococcal infection results in contamination of bedding with Cryptococcus neoformans. AB - Cryptococcus neoformans is a fungal pathogen that survives in diverse environments. To determine whether cages of mice infected with C. neoformans posed an infection risk to animal caregivers, we investigated whether the fungus could be isolated from the bedding or stool of mice infected by intratracheal (i.t.), intravenous (i.v.), or intraperitoneal (i.p.) routes. The bedding of mice infected i.t. was contaminated with C. neoformans. In contrast, no contamination of bedding with C. neoformans was detected in cages of mice infected i.v. or i.p. C. neoformans was not isolated from murine feces. The C. neoformans strain recovered from bedding material was indistinguishable from the infecting strain by biochemical and molecular techniques. This result suggests that precautions may be warranted when disposing bedding from cages that housed mice with pulmonary C. neoformans infection. PMID- 12906396 TI - Species-specific assessment of pain in laboratory animals. AB - Pain has been defined by the International Association for the Study of Pain as "an unpleasant sensory and emotional experience associated with actual or potential damage or described in terms of such damage". However, the ability to describe the concept of pain is difficult largely because pain is an individualized and subjective experience. What one person finds painful, another may not; what relieves pain for one may not do so for another. Awareness of pain management has become an important health issue for humans and animals. To effectively manage pain, it is crucial to be able to identify it, and identification of pain in animals can be especially problematic. Recognition and alleviation of pain in animals used in biomedical research and teaching is an important goal, both from a humane and regulatory perspective. This paper will: 1) review current literature regarding pain assessment using pain scales and 2) describe how an institutional care and use committee (IACUC) has implemented an effective pain scoring system to allow for an objective, accurate, and humane assessment of pain experienced by animals used in biomedical research. PMID- 12906397 TI - Postoperative pain management using fentanyl patches in dogs. AB - To the ability of the fentanyl patch to control pain in a postoperative canine model, we provided two male beagles with 25-mg/h patches and two with 50-mg/h patches 24 h prior to surgery. Each animal underwent a major abdominal surgical procedure to place three separate catheters with associated vascular access ports. Serum plasma levels of fentanyl were analyzed at multiple time points throughout the study period. Animals were subjectively assessed for postoperative pain by using a Simple Descriptive Scale at regular intervals postoperatively. Other parameters observed and recorded included heart and respiration rates, rectal temperature, appetite, and activity. The fentanyl patch appeared to adequately control postoperative pain in our canine abdominal surgical model. Three animals demonstrated mild pain 1 to 2 h postoperatively. Two animals, one from each dose group, showed mild pain 8 h postoperatively. Mild pain is commensurate with USDA category C, which encompasses procedures that do not result in more than momentary or slight pain or distress and do not require intervention. At no other time points were any of the animals considered to be in pain. Our study also suggested that increased subcutaneous fat delayed the rate of absorption of fentanyl. The lower body-weight beagles, which had the 25-mg/h patches, reached reported human serum analgesic levels within 8 h after placement, whereas the heavier beagles with the 50-mg/h patches reached human serum analgesic levels 12 h after placement. Fentanyl concentrations remained at the reported human analgesic levels in all animals between 2 to 4 h after the patches were removed. Regardless of the dose, decreases in heart rate, respiration rate, and temperature were observed in all four animals 12 h after placing the patches. Issues regarding the regulatory requirements to prevent drug abuse, the side effects and potency of fentanyl, and the prolonged duration of action as a transdermal system should be addressed by the veterinarian when considering usage of this analgesic method. PMID- 12906398 TI - The sedative and behavioral effects of nalbuphine in dogs. AB - We compared the degree of sedation and frequency and intensity of adverse behaviors in dogs associated with nalbuphine when combined with acepromazine or xylazine compared with those of acepromazine or xylazine alone. Twenty-four dogs (13 female, 11 male) undergoing routine ovariohysterectomy or castration were randomly assigned to one of four groups. Group NX received 0.5 mg/kg nalbuphine and 0.5 mg/kg xylazine subcutaneously (s.c.). Group X received 0.5 mg/kg xylazine s.c. Group NA received 0.5 mg/kg nalbuphine and 0.05 mg/kg acepromazine s.c. Group A received 0.05 mg/kg acepromazine s.c. All dogs received 0.01 mg/kg glycopyrrolate s.c. All doses were administered preoperatively. Preoperative resting measurements of heart rate, respiratory rate, rectal temperature, and body weight were obtained. Sedation was scored both inside and outside a kennel prior to drug administration and at 10, 20, and 30 min after drug administration. Dogs were assessed for behavioral responses (leg withdrawal, shivering, rigidity, orienting, panting, struggling, vocalization, wide-eyed facial expression, breath holding, salivating, hiding, biting, or requiring a muzzle) during three time periods: placing the dog on the table, clipping and prepping of forelimb, and intravenous catheterization. Postoperative recovery behaviors were scored. Expired halothane concentrations were recorded at 15, 30, and 45 min postinduction. Significant differences occurred in the level of sedation at 30 min between dogs receiving nalbuphine and xylazine or xylazine only compared with dogs receiving acepromazine. There was a significant difference in behavioral scores with respect to leg withdrawal and orienting during clipping/prepping between dogs receiving nalbuphine and xylazine compared with dogs receiving xylazine. The combination of nalbuphine and xylazine is a useful premedicant which provided greater sedation than acepromazine and reduced some anxiety behaviors more than did xylazine alone. Nalbuphine is an inexpensive opioid and currently is not a controlled substance in the U.S. PMID- 12906399 TI - Comparison of ketamine versus combination of ketamine and medetomidine in injectable anesthetic protocols: chemical immobilization in macaques and tissue reaction in rats. AB - This study compared balanced anesthesia between ketamine alone and ketamine with medetomidine and assessed the repeated intramuscular use of ketamine and its potential for tissue damage. The combination of ketamine and medetomidine was tested in newly arrived macaques undergoing a period of quarantine in an animal facility. Results indicated that the medetomidine and ketamine combination induced a deeper, more level plane of anesthesia of longer duration than did ketamine alone. Furthermore, use of the medetomidine-reversing agent, atipamezole, permitted more rapid recovery. In addition, a preliminary study in adult rats was undertaken to assess tissue damage induced by intramuscular injection of ketamine versus the combination of ketamine and medetomidine. Histological evaluation of tissue inflammation and muscle necrosis in rats indicated that the lower dose of ketamine afforded by combination with medetomidine caused markedly less damage to muscle tissue at injection sites. PMID- 12906400 TI - Effect of pair-housing on operant behavior task performance by rhesus monkeys. AB - This study evaluated the effects of pair-housing on several operant (trained) behaviors in rhesus macaques (Macaca mulatta). Sixteen young, male, individually housed rhesus monkeys (age, 2.5 to 5.5 years) performed a battery of behaviors consisting of motivation (progressive ratio, PR), short-term memory and attention (delayed matching-to-sample, DMTS), color and position discrimination (conditioned position responding, CPR), and learning (incremental repeated acquisition, IRA) tasks. Behavioral assessments occurred 5 days/week, with the PR, IRA, and CPR tasks presented on one test day, and the DMTS task presented on the next test day. Thus, each task was performed two or three days/week. Eight subjects then were pair-housed, while eight age-matched controls remained individually housed. Pair-housed monkeys were separated for behavior testing and feeding but allowed access to each other approximately 20 h/day. The performance of the two groups of monkeys were compared for the 2 months prior to pairing, for a 2-month transition period as the pairs adjusted to the new housing situation, and for a 2-month period after the pairs had been established. Performance of the CPR and IRA tasks did not change over time in either group. For the PR and DMTS tasks, the number of trials completed increased over the course of the study in the controls but not in the pair-housed monkeys. In conclusion, pair-housing monkeys is feasible for studies involving operant behavior testing as a model for a variety of complex brain functions. However, housing condition may affect some test parameters, and this must be taken into consideration during experimental design. PMID- 12906401 TI - Some observations on the pharmacological properties of ivermectin during treatment of a mite infestation in mice. AB - Ivermectin has become one of the most widely used drugs for the treatment of parasitic infections in laboratory rodents. Despite its wide popularity, little has been published regarding its pharmacokinetic properties in mice. We made use of a routine mite control treatment in a conventional animal unit to gain some knowledge of these properties. Numerous inbred, outbred, and transgenic lines of mice were being treated with ivermectin in the drinking water (32 mg/L) for a chronic Myocoptes infestation. A sample of mice from different cages and rooms was culled at various time intervals to check levels of ivermectin in blood. In addition, cellophane tape impressions of fur were obtained from mice undergoing treatment to monitor the effectiveness of ivermectin in controlling the mite infestation. Results showed that ivermectin levels rose sharply in the serum of treated mice and gradually fell once treatment was discontinued. Maximum levels detected at the end of a 10-day period of treatment were 90 ng/ml. Once treatment was discontinued, serum levels of the drug were undetectable after 7 days. In addition, treatment with ivermectin proved very effective in reducing parasite burdens in the colony. Results were in line with known pharmacological properties of ivermectin in other species. Nevertheless, it was worth noting that serum levels can vary markedly depending on various factors, something to be taken into account when considering treatment of mice, especially transgenics. PMID- 12906402 TI - Evaluation of coinfection with pinworms (Aspiculuris tetraptera, Dentostomella translucida, and Syphacia obvelata) in gerbils and mice. AB - Mongolian gerbils (Meriones unguiculatus) naturally infected with the nematode Dentostomella translucida and mice (Mus musculus) that were either pinworm-free or harboring either or both Aspiculuris tetraptera and Syphacia abvelata were crossinfected experimentally with these nematode species. Gerbils were susceptible to infections by S. obvelata and A. tetraptera from mice, whereas mice were resistant to infection by D. translucida from gerbils. The data presented are the first regarding coinfections with these three pinworms in gerbils. PMID- 12906403 TI - Inoculation of Staphylococcus xylosus in SJL/J mice to determine pathogenicity. AB - An experimental study was performed to investigate whether intradermal tail inoculations of Staphylococcus xylosus would result in pathologic lesions in the SJL/J strain of mice (Mus musculus). This organism historically has been classified as a nonpathogenic, commensal bacterium associated with skin and mucous membranes and rarely implicated in infections. In this study, SJL/J mice inoculated with S. xylosus developed cutaneous tail lesions post-inoculation, and the organism was recovered from those lesions. Inoculation was accomplished by surgically inserting silk suture impregnated with the concentrated suspension of bacteria. In addition, a superficial abrasion was created adjacent to the suture, and a bacterial suspension was applied. Approximately 80% of the mice in the inoculated groups developed dermatologic lesions, compared with 0% in the control group. Mice with lesions were treated with Sulfamethoxazole-Trimethoprim in the drinking water continuously for 28 days. For the mice assigned to the treatment group, this treatment resulted in resolution of the cutaneous tail lesions. PMID- 12906404 TI - An alternative method of chronic cerebrospinal fluid collection via the cisterna magna in conscious rhesus monkeys. AB - Models of chronic cerebrospinal fluid (CSF) collection previously have been established for nonhuman primates and canines; many of these methods implement stainless-steel cannulas into the lateral or 4th ventricles or catheters into the cerebral or spinal subarachnoid space. These models have proved successful and reliable but unfortunately require invasive techniques to pass through the skull or require a laminectomy to enter the spinal subarachnoid space, involve the use of expensive and highly specialized stereotaxic equipment for the precise placement of the implants, and may require exteriorized hardware which is cumbersome to maintain and unaesthetic. The model we developed for the rhesus monkey allows for direct access to CSF outflow from the cisterna magna by using a 3.5-French fenestrated silicone catheter which was placed 1.0 cm into the cisterna. The catheter was attached to a titanium port placed subcutaneously between the scapulae to permit easy access for sampling CSF in a conscious, chaired rhesus monkey. We currently have instrumented animals from which we have consistently collected CSF for over 18 months. This novel, economical, less invasive method permits chronic, reliable collection of CSF in conscious rhesus monkeys and has the additional advantages that the model is easier to maintain and more aesthetic. PMID- 12906405 TI - Stochastic annealing. AB - We show how to simulate a system in thermal equilibrium when the energy cannot be evaluated exactly: the error distribution needs to be symmetric, but it does not need to be known. We also solve the Ceperley-Dewing version of this problem, where the error distribution is taken to be fully known. These underlying ideas give an effective optimization strategy for problems where the evaluation of each design can be sampled only statistically, including an application to protein folding. PMID- 12906406 TI - Derivation of the Gross-Pitaevskii equation for condensed bosons from the Bogoliubov-de Gennes equations for superfluid fermions. AB - We derive the time-independent Gross-Pitaevskii equation at zero temperature for condensed bosons, which form as bound-fermion pairs when the mutual fermionic attractive interaction is sufficiently strong, from the strong-coupling limit of the Bogoliubov-de Gennes equations that describe superfluid fermions in the presence of an external potential. Three-body corrections to the Gross-Pitaevskii equation are also obtained by our approach. Our results are relevant to the recent advances with ultracold fermionic atoms in a trap. PMID- 12906407 TI - Quantum Hall fractions in rotating Bose-Einstein condensates. AB - We study the quantum Hall phases that appear in the dilute limit of rotating Bose Einstein condensates. By exact diagonalization in a spherical geometry we obtain the ground state and low-lying excited states of a small number of bosons as a function of the filling fraction nu, the ratio of the number of bosons to the number of vortices. We show the occurrence of the Jain principal sequence of incompressible liquids for nu=2/3,3/4,4/3,5/4 in addition to the Laughlin state nu=1/2 as well as the Pfaffian state for nu=1. We give gap estimates by finite size scaling of both charged and neutral excitations. PMID- 12906408 TI - Critical dynamics of model C resolved. AB - We analyze the field theoretic functions of the dynamical model C in two-loop order. Our results correct long-standing errors in these functions published by several authors. We discuss, in particular, the fixed points for the ratio w* of the two time scales involved, as well as their stability. The regions of the "phase diagram," whose axes are the spatial dimension d and number of order parameter components n, correspond to these fixed points; previous authors have found, in addition, an anomalous region in which the scaling properties were unclear. We show that such an anomalous region does not exist. There are only two regions: one with a finite fixed-point w* where the dynamical exponent z=2+alpha/nu, and another where w*=0 and z is equal to the model A value. We show how the one-loop result is recovered from the two-loop result in the limit epsilon=4-d going to zero. PMID- 12906409 TI - Persistence of manifolds in nonequilibrium critical dynamics. AB - We study the persistence probability P(t) that, starting from a random initial condition, the magnetization of a d'-dimensional manifold of a d-dimensional spin system at its critical point does not change sign up to time t. For d'>0 we find three distinct late-time decay forms for P(t): exponential, stretched exponential, and power law, depending on a single parameter zeta=(D-2+eta)/z, where D=d-d' and eta,z are standard critical exponents. In particular, we predict that for a line magnetization in the critical d=2 Ising model, P(t) decays as a power law while, for d=3, P(t) decays as a power of t for a plane magnetization but as a stretched exponential for a line magnetization. Numerical results are consistent with these predictions. PMID- 12906410 TI - Measurement of decay parameters for Xi- --> Lambda pi- decay. AB - Based on 1.35 x 10(6) polarized Xi(-) events, we measure the parameter phi(Xi) to be -1.61 degrees +/-2.66 degrees +/-0.37 degrees for the Xi(-)-->Lambda pi(-) decay. New results for the parameters beta(Xi) and gamma(Xi) are also presented. Assuming that the CP-violating phase difference is negligible, we deduce the strong phase difference between the P-wave and S-wave amplitudes of the Lambda pi final state to be 3.17 degrees +/-5.28 degrees +/-0.73 degrees, reducing the uncertainty in estimating the level of CP violation in Xi-hyperon decay. PMID- 12906411 TI - Vacuum photon splitting in Lorentz-violating quantum electrodynamics. AB - Radiative corrections arising from Lorentz violation in the fermion sector induce a nonzero amplitude for vacuum photon splitting. At one loop, the on-shell amplitude acquires both CPT-even and CPT-odd contributions forbidden in conventional electrodynamics. PMID- 12906412 TI - Breached pairing superfluidity: possible realization in QCD. AB - We propose a wide universality class of gapless superfluids, and analyze a limit that might be realized in quark matter at intermediate densities. In the breached pairing color superconducting phase heavy s quarks, with a small Fermi surface, pair with light u or d quarks. The ground state has a superfluid and a normal Fermi component simultaneously. We expect a second-order phase transition, as a function of increasing density, from the breached pairing phase to the conventional color-flavor locked phase. PMID- 12906413 TI - QCD inequalities for the nucleon mass and the free energy of baryonic matter. AB - The positivity of the integrand of certain Euclidean space functional integrals for two flavor QCD with degenerate quark masses implies that the free energy per unit volume for QCD with a baryon chemical potential mu(B) (and zero isospin chemical potential) is greater than the free energy with an isospin chemical potential mu(I)=(2 mu(B)/N(c)) (and zero baryon chemical potential). The same result applies to QCD with any number of heavy flavors in addition to the two light flavors so long as the chemical potential is understood as applying to the light quark contributions to the baryon number. This relation implies a bound on the nucleon mass: there exists a particle X in QCD (presumably the pion) such that M(N)> or =(N(c) m(X)/2 I(X)) where m(X) is the mass of the particle and I(X) is its isospin. PMID- 12906414 TI - Dispersion energy from density-functional theory description of monomers. AB - A method is proposed for calculations of dispersion energy at finite intermonomer separations. It uses a generalized Casimir-Polder formula evaluated with dynamic density susceptibilities provided by time-dependent density-functional theory. The method recovers the dispersion energies of He, Ne, and H2O dimers to within 3% or better. Since the computational effort of the new algorithm scales approximately as the third power of system size, the method is much more efficient than standard wave-function methods capable of predicting the dispersion energy at a similarly high level of accuracy. PMID- 12906415 TI - Self-organized periodic photonic structure in a nonchiral liquid crystal. AB - A hybrid-aligned cell of the smectic A liquid material 8CB gives two stable director configurations, one of which is periodic and gives strong diffraction of light. This photonic lattice director profile arises from "frustration" caused by conflicting constraints imposed by the boundary conditions and the constant amplitude smectic density wave. A model is proposed which accords well with the experiment results, predicting correctly the dependence of the periodicity on the cell thickness and reproducing optical polarization microscopy results. PMID- 12906416 TI - Directional tunneling escape from nearly spherical optical resonators. AB - We report the surprising observation of directional tunneling escape from nearly spherical fused-silica optical resonators, in which most of the phase space is filled with nonchaotic regular trajectories. Experimental and theoretical studies of the dependence of the far-field emission pattern on both the degree of deformation and the excitation condition show that nonperturbative phase-space structures in the internal ray dynamics profoundly affect tunneling leakage of the whispering-gallery modes. PMID- 12906417 TI - Characterizing individual scattering events by measuring the amplitude and phase of the electric field diffusing through a random medium. AB - We describe observations of the amplitude and phase of an electric field diffusing through a three-dimensional random medium, using terahertz time-domain spectroscopy. These measurements are spatially resolved with a resolution smaller than the speckle spot size and temporally resolved with a resolution better than one optical cycle. By computing correlation functions between fields measured at different positions and with different temporal delays, it is possible to obtain information about individual scattering events experienced by the diffusing field. This represents a new method for characterizing a multiply scattered wave. PMID- 12906418 TI - Chaotic dynamics of the fractional Lorenz system. AB - In this Letter we introduce a generalization of the Lorenz dynamical system using fractional derivatives. Thus, the system can have an effective noninteger dimension Sigma defined as a sum of the orders of all involved derivatives. We found that the system with Sigma<3 can exhibit chaotic behavior. A striking finding is that there is a critical value of the effective dimension Sigma(cr), under which the system undergoes a transition from chaotic dynamics to regular one. PMID- 12906419 TI - On-off collective imperfect phase synchronization and bursts in wave energy in a turbulent state. AB - A new type of synchronization, on-off collective imperfect phase synchronization, is found in a turbulent state. In the driver frame the nonlinear wave system can be transformed to a set of coupled oscillators moving in a potential related to the unstable steady wave. In "on" stages the oscillators in different spatial scales adjust themselves to collective imperfect phase synchronization, inducing strong bursts in the wave energy. The interspike intervals display a power-law distribution. In addition to the embedded saddle point, it is emphasized that the delocalization of the master mode also plays an important role in developing the on-off synchronization. PMID- 12906420 TI - Two-dimensional turbulence of dilute polymer solutions. AB - We investigate theoretically and numerically the effect of polymer additives on two-dimensional turbulence by means of a viscoelastic model. We provide compelling evidence that, at vanishingly small concentrations, such that the polymers are passively transported, the probability distribution of polymer elongation has a power law tail: Its slope is related to the statistics of finite time Lyapunov exponents of the flow, in quantitative agreement with theoretical predictions. We show that at finite concentrations and sufficiently large elasticity the polymers react on the flow with manifold consequences: Velocity fluctuations are drastically depleted, as observed in soap film experiments; the velocity statistics becomes strongly intermittent; the distribution of finite time Lyapunov exponents shifts to lower values, signaling the reduction of Lagrangian chaos. PMID- 12906421 TI - Observation of higher-order snake resonances in polarized proton acceleration in RHIC. AB - Higher-order and coupled snake resonances were observed during the 2002 polarized proton run in RHIC. Strong depolarization was observed when the fractional part of the vertical betatron tune approached 1/4, and when the fractional part of the horizontal tune approached 3/14. Because of the closed orbit error, each snake resonance splits into two. From the width of the observed snake resonances, we can derive the strength of the imperfection spin resonance. Our results appear consistent with the measured closed orbit error. PMID- 12906422 TI - Turbulent particle transport in magnetized plasmas. AB - Particle transport in magnetized plasmas is investigated with a fluid model of drift wave turbulence. An analytical calculation shows that magnetic field curvature and thermodiffusion drive an anomalous pinch. The curvature driven pinch velocity is consistent with the prediction of turbulence equipartition theory. The thermodiffusion flux is found to be directed inward for a small ratio of electron to ion pressure gradient, and it reverses its sign when increasing this ratio. Numerical simulations confirm that a turbulent particle pinch exists. It is mainly driven by curvature for equal ion and electron heat sources. The sign and relative weights of the curvature and thermodiffusion pinches are consistent with the analytical calculation. PMID- 12906423 TI - Nonequilibrium and non-steady-state evolution of a shock state. AB - Using numerical simulations, we study the evolution of silicon in the passage of a constant-pressure shock wave launched from an adjacent pusher. We examine also its optical characteristics of reflectivity and emission. Our finding points to the study of shocked interfaces as a novel means to explore nonequilibrium, non steady-state behaviors of shock states and an alternative approach to assess electron-ion equilibration rate in a shock wave. It also reveals important structures in such a shock wave in contrast to its usual notion as a propagating discontinuity. This offers some possibilities for reconciling the different findings on the compressibility of deuterium. PMID- 12906424 TI - Resistive collimation of electron beams in laser-produced plasmas. AB - Intense relativistic electron beams, produced by high-intensity short-pulse laser irradiation of a solid target, have many potential applications including fusion by fast ignition. Using a unique Fokker-Planck code, supported by analytic calculations, we show that fast electrons can be collimated into a beam even when the fast electron source is not strongly anisotropic, and we derive a condition for collimation to occur. PMID- 12906425 TI - Laboratory realization of an Alfven wave maser. AB - It is demonstrated that a frequency selective Alfven-wave resonator can be realized by applying a nonuniform magnetic field to a plasma region bounded between a cathode and a semitransparent mesh anode. When a current threshold is exceeded, selective amplification results in a highly coherent, large amplitude wave that propagates into an adjacent plasma column. PMID- 12906426 TI - What is the ground-state structure of the thinnest Si nanowires? AB - Pristine silicon whiskers are compared through energy analysis by separating the surface, edge, and bulk contributions, and by energy computation for a variety of structures and diameters d. It is shown that for d<6 nm a polycrystalline wire of five-fold symmetry, rather than single-crystal types, represents the ground state. It remains stable in molecular dynamics tests up to approximately 1000 K. Its specific surface reconstruction also stands out in that it favors kinetics of whisker growth and thus appears potentially realizable. PMID- 12906427 TI - Shock-induced transformation of Al2O3 and LiF into semiconducting liquids. AB - Shock compression of sapphire (Al2O3) and lithium fluoride (LiF) to pressures above 5 Mbar has been observed to transform these transparent, wide band-gap insulators into partially degenerate liquid semiconductors with optical reflectivities of several percent. Reflectivities rise steadily with shock pressure up to 45% in sapphire at 20 Mbar and 20% in LiF at 13 Mbar. Using a simple model, the electron scattering length was inferred to be approximately the interatomic distance. In addition, several equation-of-state points at these pressures were measured. PMID- 12906428 TI - Adsorption of argon on carbon nanotube bundles and its influence on the bundle lattice parameter. AB - We report experimental studies of the adsorption characteristics and structure of both 36Ar and 40Ar on single-wall carbon nanotube bundles. The structural studies make use of the large difference in coherent neutron scattering cross section for the two Ar isotopes to explore the influence of the adsorbate on the nanotube lattice parameter. We observe no dilation of the nanotube lattice with 40Ar, and explain the apparent expansion of this lattice upon 36Ar adsorption by the location of the adsorbed Ar atoms on the outer bundle surface. PMID- 12906429 TI - Mg segregation at Al/Al3Sc heterophase interfaces on an atomic scale: experiments and computations. AB - Microscopic factors governing solute partitioning in ternary two-phase Al-Sc-Mg alloys are investigated combining three-dimensional-atom-probe (3DAP) microscopy measurements with first-principles computations. 3DAP is employed to measure composition profiles with subnanometer-scale resolution, leading to the identification of a large enhancement of Mg solute at the coherent alpha Al/Al(3)Sc (fcc/L1(2)) heterophase interface. First-principles calculations establish an equilibrium driving force for this interfacial segregation reflecting the nature of the interatomic interactions. PMID- 12906430 TI - Dimensional crossover and charge order in half-doped manganites and cobaltites. AB - We propose a generic model for understanding the effect of quenched disorder on charge-ordering in half-doped manganese and cobalt oxides with different crystal structures. Current experimental observations are explained in the light of the global phase diagram of the model. PMID- 12906432 TI - Dynamic nuclear polarization by electrical spin injection in ferromagnet semiconductor heterostructures. AB - Electrical spin injection from Fe into AlxGa1-xAs quantum well heterostructures is demonstrated in small (<500 Oe) in-plane magnetic fields. The measurement is sensitive only to the component of the spin that precesses about the internal magnetic field in the semiconductor. This field is much larger than the applied field and depends strongly on the injection current density. Details of the observed hysteresis in the spin injection signal are reproduced in a model that incorporates the magnetocrystalline anisotropy of the epitaxial Fe film, spin relaxation in the semiconductor, and the dynamic polarization of nuclei by the injected spins. PMID- 12906431 TI - Probing the different stages in contacting a single molecular wire. AB - A crucial problem in molecular electronics is the control of the electronic contact between a molecule and its electrodes. As a model system, we investigated the contact between the molecular wire group of a C90H98 (Lander) molecule and the edge of a Cu(111) monatomic step. The reproducible contact and decontact of the wire was obtained by manipulating the Lander with a low temperature scanning tunneling microscope. The electronic standing wave patterns on the Cu(111) surface serve to monitor the local electronic perturbation caused by the interaction of the wire end with the step edge, giving information on the quality of the contact. PMID- 12906433 TI - Enhanced electrical resistivity before Neel order in the metals RCuAs2 (R=Sm, Gd, Tb, and Dy). AB - We report an unusual temperature (T) dependent electrical resistivity (rho) behavior in a class of ternary intermetallic compounds of the type RCuAs2 (R=rare earths). For some rare earths (Sm, Gd, Tb, and Dy) with negligible 4f hybridization, there is a pronounced minimum in rho(T) far above respective Neel temperatures (T(N)). However, for the rare earths which are more prone to exhibit such a rho(T) minimum due to 4f-covalent mixing and the Kondo effect, this minimum is depressed. These findings, difficult to explain within the hitherto known concepts, present an interesting scenario in magnetism. PMID- 12906434 TI - Fractional statistics in the fractional quantum Hall effect. AB - A microscopic confirmation of the fractional statistics of the quasiparticles in the fractional quantum Hall effect has so far been lacking. We calculate the statistics of the composite-fermion quasiparticles at nu=1/3 and nu=2/5 by evaluating the Berry phase for a closed loop encircling another composite-fermion quasiparticle. A careful consideration of subtle perturbations in the trajectory due to the presence of an additional quasiparticle is crucial for obtaining the correct value of the statistics. The conditions for the applicability of the fractional statistics concept are discussed. PMID- 12906435 TI - Field theoretical description of quantum Hall edge reconstruction. AB - We propose a generalization of the chiral Luttinger liquid theory to allow for a unified description of quantum Hall edges with or without edge reconstruction. Within this description edge reconstruction is found to be a quantum phase transition in the universality class of a one-dimensional dilute Bose gas transition, whose critical behavior can be obtained exactly. At principal filling factors nu=1/m, we show the additional edge mode due to edge reconstruction modifies the point contact tunneling exponent in the low-energy limit, by a small and nonuniversal amount. PMID- 12906436 TI - Signature of a chemical bond in the conductance between two metal surfaces. AB - Conductance in monatomic metal contacts is quantized; it increases in discrete steps of one conductance quantum 2e(2)/h. By contrast, in a vacuum barrier between two metal surfaces we find that conductance increases linearly and continuously with the interaction energy between individual atoms. This behavior shows unambiguously that current flow between single atoms is a measure for their chemical interaction. In the controlled environment of a scanning tunneling microscope it should allow us to study the formation of covalent bonds up to the point where these atoms finally jump into contact. PMID- 12906437 TI - Phase-sensitive noise in quantum dots under periodic perturbation. AB - We evaluate the ensemble averaged noise in a chaotic quantum dot subject to dc bias and a periodic perturbation of frequency Omega. The noise displays cusps at bias V(n)=n variant Planck's over 2 pi Omega/e that survive the average, even when the period of the perturbation is far shorter than the dwell time in the dot. These features are sensitive to the phase of the time-dependent scattering amplitudes of electrons to pass through the system, and thus provide a novel signature of phase-coherent transport that persists into the nonadiabatic limit. PMID- 12906438 TI - Anisotropy in conductance of a quasi-one-dimensional metallic surface state measured by a square micro-four-point probe method. AB - We have devised a "square micro-four-point probe method" using an independently driven ultrahigh-vacuum four-tip scanning tunneling microscope, and succeeded for the first time to directly measure anisotropic electrical conductance of a single atomic layer on a solid surface. A quasi-one-dimensional metal of a single-domain Si(111)4 x 1-In had a surface-state conductance along the metallic atom chains (sigma(axially)) to be 7.2(+/-0.6) x 10(-4) S/square at room temperature, which was larger than that in the perpendicular direction (sigma(radially)) by approximately 60 times. The sigma(axially) was consistently interpreted by a Boltzmann equation with the anisotropic surface-state band dispersion, while the sigma(radially) was dominated by a surface-space-charge-layer conductance. PMID- 12906439 TI - Evidence for two types of low-energy charge transfer excitations in Sr2CuO3. AB - A comparative analysis of electron energy-loss spectroscopy (EELS) spectra for the 1D insulating cuprate Sr2CuO3 with transferred momentum q--> axially and radially to the chain axis allows one to elucidate the structure of the charge transfer gap in in-chain response. It is determined by the superposition of two types of excitations with different magnitudes of dispersion. The low-energy response with q--> radially to the chain direction, but yet within the plane of CuO4 plaquettes, exhibits also a dispersionless peak near 2 eV. The theoretical simulation of the EELS data using exact diagonalizations of an appropriate extended Hubbard Hamiltonian for relevant clusters requires the explicit consideration of low-lying oxygen 2p pi states within the CuO4 plaquette plane beyond the standard pd sigma extended Hubbard model widely used for cuprates with corner-shared CuO4 plaquettes. PMID- 12906440 TI - Test for pairing symmetry based on spin fluctuations in the organic superconductor kappa-(BEDT-TTF)2X. AB - We propose that the superconducting pairing symmetry of organic superconductors kappa-(BEDT-TTF)2X can be determined by measuring the position in momentum space of the incommensurate peaks of the spin susceptibility. Using the weak coupling BCS theory and including the many-body effects via the random-phase approximation for the Hubbard model on an anisotropic triangular lattice, we show that the position of these peaks is uniquely determined by the pairing symmetry of the superconducting state and the geometry of the Fermi surface. We demonstrate the different incommensurate patterns of spin responses for d(x(2)-y(2-)) and d(xy) like pairing states. In addition, we find that there is no spin resonance mode in the reasonable range of parameters discussed. PMID- 12906441 TI - Determination of the Fermi surface of MgB2 by the de Haas-van Alphen effect. AB - We report measurements of the de Haas-van Alphen (dHvA) effect for single crystals of MgB2, in magnetic fields up to 32 T. In contrast to our earlier work, dHvA orbits from all four sheets of the Fermi surface were detected. Our results are in good overall agreement with calculations of the electronic structure and the electron-phonon mass enhancements of the various orbits, but there are some small quantitative discrepancies. In particular, systematic differences in the relative volumes of the Fermi-surface sheets and the magnitudes of the electron phonon coupling constants could be large enough to affect detailed calculations of T(c) and other superconducting properties. PMID- 12906442 TI - c-Axis optical sum rule and a possible new collective mode in La2-xSrxCuO4. AB - We present the c-axis optical conductivity sigma(1c)(omega,T) of underdoped (x=0.12) and optimally doped (x=0.15) La2-xSrxCuO4 from 4 meV to 1.8 eV obtained by a combination of reflectivity and transmission spectra. In addition to the opening of the superconducting gap, we observe an increase of conductivity above the gap up to 270 meV with a maximal effect at about 120 meV. This may indicate a new collective mode at a surprisingly large energy scale. The Ferrell-Glover Tinkham sum rule is violated for both doping levels. Although the relative value of the violation is much larger for the under-doped sample, the absolute increase of the low-frequency spectral weight, including that of the condensate, is higher in the optimally doped regime. Our results resemble in many respects the observations in YBa(2)Cu(3)O(7-delta). PMID- 12906443 TI - Numerical simulation evidence of dynamical transverse meissner effect and moving bose glass phase. AB - We present 3D numerical simulation results of moving vortex lattices in the presence of 1D correlated disorder at zero temperature. Our results with field tilting confirm the theoretical predictions of a moving Bose glass phase, characterized by transverse pinning and dynamical transverse Meissner effect, the moving flux lines being localized along the correlated disorder direction. Beyond a critical transverse field, vortex lines exhibit along all their length a "kink" structure resulting from an effective static "tin roof" pinning potential in the transverse direction. PMID- 12906444 TI - Large negative magnetic contribution to the thermal expansion in iron-platinum alloys: quantitative theory of the Invar effect. AB - We show that the large negative magnetic contribution to the thermal expansion in disordered Fe-Pt alloys can be understood within the disordered local moment (DLM) approach. On the basis of first principles calculations we quantitatively describe the spontaneous volume magnetostriction for various Pt concentrations. It is found that the Invar effect in these alloys is entirely related to the state of thermal magnetic disorder modeled by the DLM states. We also show that the experimentally observed anomaly in the temperature dependence of the magnetization is due to a spontaneous reduction of the local magnetic moments rather than to "hidden excitations." PMID- 12906445 TI - Magnetic superlattices with variable interlayer exchange coupling: a new approach for the investigation of low-dimensional magnetism. AB - We have investigated the magnetic order in an [Fe(2)/(VHx)(13)] x 200 superlattice as a function of temperature and hydrogen content in the vanadium layers. A J(radially)-T magnetic phase diagram was established where J(radially) denotes the interlayer exchange coupling between adjacent Fe planes. We propose that Fe/V superlattices, in which the ratio of interlayer to intralayer coupling can be tuned continuously and reversibly via hydrogen in the nonmagnetic vanadium, offer a new approach for the study of low-dimensional magnetism and crossover effects near the transition from ferromagnetic to antiferromagnetic order. PMID- 12906446 TI - Full aging in spin glasses. AB - The discovery of dynamic memory effects in the magnetization decays of spin glasses in 1983 marked a turning point in the study of the highly disordered spin glass state. Detailed studies of the memory effects have led to much progress in understanding the qualitative features of the phase space. Even so, the exact nature of the magnetization decay functions has remained elusive, causing confusion. In this Letter, we report strong evidence that the thermoremanent magnetization decays scale with the waiting time t(w). By employing a series of cooling protocols, we demonstrate that the rate at which the sample is cooled to the measuring temperature plays a major role in the determination of scaling. As the effective cooling time t(eff)(c) decreases, t/t(w) scaling improves and for t(eff)(c)<20 s we find almost perfect t/t(w) scaling, i.e., full aging. PMID- 12906447 TI - Half-metallic ferromagnetism and structural stability of zincblende phases of the transition-metal chalcogenides. AB - An accurate density-functional method is used to study systematically half metallic ferromagnetism and stability of zincblende phases of 3d-transition-metal chalcogenides. The zincblende CrTe, CrSe, and VTe phases are found to be excellent half-metallic ferromagnets with large half-metallic gaps (up to 0.88 eV). They are mechanically stable and approximately 0.31-0.53 eV per formula unit higher in total energy than the corresponding nickel-arsenide ground-state phases, and therefore would be grown epitaxially in the form of films and layers thick enough for spintronic applications. PMID- 12906448 TI - Extended quantum critical phase in a magnetized spin-1/2 antiferromagnetic chain. AB - Measurements are reported of the magnetic field dependence of excitations in the quantum critical state of the spin S=1/2 linear chain Heisenberg antiferromagnet copper pyrazine dinitrate (CuPzN). The complete spectrum was measured at k(B)T/J< or =0.025 for H=0 and H=8.7 T, where the system is approximately 30% magnetized. At H=0, the results are in agreement with exact calculations of the dynamic spin correlation function for a two-spinon continuum. At H=8.7 T, there are multiple overlapping continua with incommensurate soft modes. The boundaries of these continua confirm long-standing predictions, and the intensities are consistent with exact diagonalization and Bethe ansatz calculations. PMID- 12906449 TI - Spectral weight contributions of many-particle bound States and continuum. AB - Cluster expansion methods are developed for calculating the spectral weight contributions of multiparticle excitations--continuum and bound states--to high orders. A complete 13th order calculation is carried out for the alternating Heisenberg chain. For lambda=0.27, relevant to the material Cu(NO3)(2).2.5D(2)O, we present detailed spectral weights for the two-triplet continuum and all bound states. We also examine the variation of the relative weights of one- and two particle states with bond alternation from the dimerized to the uniform chain limit. PMID- 12906450 TI - Oscillatory interlayer exchange coupling and its temperature dependence in [Pt/Co]3/NiO/[Co/Pt]3 multilayers with perpendicular anisotropy. AB - Interlayer exchange coupling that oscillates between antiferromagnetic and ferromagnetic as a function of NiO thickness has been observed in [Pt(5 A)/Co(4 A)](3)/NiO(t(NiO) A)/[Co(4 A)/Pt(5 A)](3) multilayers with out-of-plane anisotropy. The period of oscillation corresponds to approximately 2 monolayers of NiO. This oscillatory behavior is possibly attributed to the antiferromagnetic ordering in NiO. The antiferromagnetic interlayer exchange coupling for the 11 A NiO layer shows an increase in coupling strength with increasing temperature, in agreement with the quantum interference model of Bruno for insulating spacer layers. A coexistence of exchange biasing and antiferromagnetic interlayer exchange coupling has been observed below T=250 K. PMID- 12906451 TI - Ferromagnetic ordering in alkali-metal iron antimonides: NaFe4Sb12 and KFe4Sb12. AB - New alkali-metal compounds with the filled-skutterudite structure were synthesized and their chemical and physical properties investigated. X-ray diffraction, microprobe, and chemical analysis established the structure and the composition without defects on the cation site. Magnetization, ac susceptibility, specific heat, resistivity, and NMR or NQR demonstrated NaFe4Sb12 to be ferromagnetic below approximately 85 K and to exhibit an additional magnetic anomaly around 40 K. Band structure calculations find a large density of states at the Fermi energy and a ferromagnetic ground state. Similar behavior was observed for KFe4Sb12. PMID- 12906452 TI - Extended dynamical mean field theory study of the periodic Anderson model. AB - We investigate the competition of the Kondo and the RKKY interactions in heavy fermion systems. We solve a periodic Anderson model using extended dynamical mean field theory (EDMFT) with quantum Monte Carlo method. We monitor simultaneously the evolution of the electronic and magnetic properties. As the RKKY coupling increases the heavy fermion quasiparticle unbinds and a local moment forms. At a critical RKKY coupling there is an onset of magnetic order. Within EDMFT the two transitions occur at different points and the disappearance of the magnetism is not described by a local quantum critical point. PMID- 12906453 TI - Nonlinear properties of left-handed metamaterials. AB - We analyze the properties of microstructured materials with negative refraction, the so-called left-handed metamaterials. We consider a two-dimensional periodic structure created by arrays of wires and split-ring resonators embedded into a nonlinear dielectric, and calculate the effective nonlinear electric permittivity and magnetic permeability. We demonstrate that the hysteresis-type dependence of the magnetic permeability on the field intensity allows changing the material properties from left- to right-handed and back. These effects can be treated as the second-order phase transitions in the transmission properties induced by the variation of an external field. PMID- 12906454 TI - Information-theoretic approach to single-particle and two-particle interference in multipath interferometers. AB - We propose entropic measures for the strength of single-particle and two-particle interference in interferometric experiments where each particle of a pair traverses a multipath interferometer. Optimal single-particle interference excludes any two-particle interference, and vice versa. We report an inequality that states the compromises allowed by quantum mechanics in intermediate situations, and identify a class of two-particle states for which the upper bound is reached. Our approach is applicable to symmetric two-partite systems of any finite dimension. PMID- 12906455 TI - Separable States can be used to distribute entanglement. AB - We show that no entanglement is necessary to distribute entanglement; that is, two distant particles can be entangled by sending a third particle that is never entangled with the other two. Similarly, two particles can become entangled by continuous interaction with a highly mixed mediating particle that never itself becomes entangled. We also consider analogous properties of completely positive maps, in which the composition of two separable maps can create entanglement. PMID- 12906456 TI - Deterministic linear optics quantum computation with single photon qubits. AB - We suggest an efficient scheme for quantum computation with linear optical elements, where the qubits are encoded in single photon states. The scheme reduces the resources required per logical gate by several orders of magnitude, compared to an earlier proposal of Knill, Laflamme, and Milburn, while the resource overhead per gate is independent of the length of the computation. A central feature of the scheme, enabling these improvements, is the prior construction of a "linked" photon state designed according to the particular quantum circuit one wishes to process. Once this state has been successfully prepared, the computation is pursued deterministically by a sequence of teleportation steps. PMID- 12906457 TI - Algorithmic cooling in a momentum state quantum computer. AB - We describe a quantum computer based upon the coherent manipulation of two-level atoms between discrete one-dimensional momentum states. Combinations of short laser pulses with kinetic energy dependent free phase evolution can perform the logical invert, exchange, controlled-NOT, and Hadamard operations on any qubits in the binary representation of the momentum state, as well as conditional phase inversion. These allow a binary right rotation, which halves the momentum distribution in a single coherent process. Fields for the coherent control of atomic momenta may thus be designed as quantum algorithms. PMID- 12906458 TI - Brownian particle in an optical potential of the washboard type. AB - Experimental observations of the fluctuation-driven net transport of silica microspheres are presented in a two-dimensional optical potential of circular symmetry created by a Bessel light beam. The optical field is tailored to break symmetry and create a static tilted periodic (washboard) potential. As the tilt of potential exceeds a threshold, a transition between locked and running modes is observed. The running mode manifests itself by the rapid accumulation of particles at the beam center. PMID- 12906459 TI - DNA dynamics in a microchannel. AB - An extended Brownian dynamics simulation method is used to characterize the dynamics of long DNA molecules flowing in microchannels. The relaxation time increases due to confinement in agreement with scaling predictions. During flow the molecules migrate toward the channel center line, and thereby segregate according to molecular weight. Capturing these effects requires the detailed incorporation of solvent flow in the simulation method, demonstrating the importance of hydrodynamic effects in the dynamics of confined macromolecules. PMID- 12906460 TI - Single molecule lifetime fluctuations reveal segmental dynamics in polymers. AB - We present a single molecule fluorescence study that allows one to probe the nanoscale segmental dynamics in amorphous polymer matrices. By recording single molecular lifetime trajectories of embedded fluorophores, peculiar excursions towards longer lifetimes are observed. The asymmetric response is shown to reflect variations in the photonic mode density as a result of the local density fluctuations of the surrounding polymer. We determine the number of polymer segments involved in a local segmental rearrangement volume around the probe. A common decrease of the number of segments with temperature is found for both investigated polymers, poly(styrene) and poly(isobutylmethacrylate). Our novel approach will prove powerful for the understanding of the nanoscale rearrangements in functional polymers. PMID- 12906461 TI - Confinement-induced colloidal attractions in equilibrium. AB - The Poisson-Boltzmann theory for colloidal electrostatic interactions predicts that charged colloidal spheres dispersed in water should repel each other, even when confined by charged surfaces. Direct measurements on highly charged polystyrene spheres, however, reveal strong, long-ranged confinement-induced attractions that have yet to be explained. We demonstrate that anomalous attractions also characterize the equilibrium pair potential for more weakly charged colloidal silica spheres sedimented into a monolayer above a glass surface. This observation substantially expands the range of conditions for which mean-field theory incorrectly predicts the sign of macroions' interactions, and provides new insights into how confinement induces long-ranged like-charge attractions. PMID- 12906462 TI - Cho and Pak reply to 'Comment on "A convergent series for the QED effective action"'. PMID- 12906463 TI - Comment on "Tracer diffusion in a dislocated lamellar system". PMID- 12906465 TI - Modern Michelson-Morley experiment using cryogenic optical resonators. AB - We report on a new test of Lorentz invariance performed by comparing the resonance frequencies of two orthogonal cryogenic optical resonators subject to Earth's rotation over approximately 1 yr. For a possible anisotropy of the speed of light c, we obtain Delta(theta)c/c(0)=(2.6+/-1.7)x10(-15). Within the Robertson-Mansouri-Sexl (RMS) test theory, this implies an isotropy violation parameter beta-delta-1 / 2=(-2.2+/-1.5)x10(-9), about 3 times lower than the best previous result. Within the general extension of the standard model of particle physics, we extract limits on seven parameters at accuracies down to 10(-15), improving the best previous result by about 2 orders of magnitude. PMID- 12906466 TI - Measurement of the interaction energy near a Feshbach resonance in a 6Li Fermi gas. AB - We investigate the strongly interacting regime in an optically trapped 6Li Fermi mixture near a Feshbach resonance. The resonance is found at 800(40) G in good agreement with theory. Anisotropic expansion of the gas is interpreted by collisional hydrodynamics. We observe an unexpected and large shift (80 G) between the resonance peak and both the maximum of atom loss and the change of sign of the interaction energy. PMID- 12906467 TI - Gravitational waves from sub-lunar-mass primordial black-hole binaries: a new probe of extradimensions. AB - In many brane world models, gravity is largely modified at the electroweak scale approximately 1 TeV. In such models, primordial black holes (PBHs) with a lunar mass M approximately 10(-7)M([circle dot]) might have been produced when the temperature of the Universe was at approximately 1 TeV. If a significant fraction of the dark halo of our galaxy consists of these lunar mass PBHs, a huge number of BH binaries will exist in our neighborhood. Third generation detectors such as EURO can detect gravitational waves from these binaries, and can also determine their chirp mass. With a new detector designed to be sensitive at high frequency bands greater, similar 1 kHz, the existence of extradimensions could be confirmed. PMID- 12906468 TI - Primordial gravity waves and weak lensing. AB - Inflation produces a primordial spectrum of gravity waves in addition to the density perturbations which seed structure formation. We compute the signature of these gravity waves in the large scale shear field. The shear can be divided into a gradient mode (G or E) and a curl mode (C or B). The latter is produced only by gravity waves, so the observations of a nonzero curl mode could be seen as evidence for inflation. We find that the expected signal from inflation is small, peaking on the largest scales at l(l+1)C(l)/2pi<10(-11) at l=2 and falling rapidly thereafter. Even for an all-sky deep survey, this signal would be below noise at all multipoles. PMID- 12906469 TI - Black holes in Godel universes and pp waves. AB - We find exact solutions for rotating and nonrotating neutral black holes in the Godel universe of five-dimensional minimal supergravity theory. We also describe the embedding of this solution in M-theory. After dimensional reduction and T duality, we obtain a supergravity solution corresponding to placing a black string in a pp-wave background. PMID- 12906471 TI - Observation of a near-threshold enhancement in the pp mass spectrum from radiative J/psi-->gammapp decays. AB - We observe a narrow enhancement near 2m(p) in the invariant mass spectrum of pp pairs from radiative J/psi-->gammapp decays. No similar structure is seen in J/psi-->pi(0)pp decays. The results are based on an analysis of a 58 x 10(6) event sample of J/psi decays accumulated with the BESII detector at the Beijing electron-positron collider. The enhancement can be fit with either an S- or P wave Breit-Wigner resonance function. In the case of the S-wave fit, the peak mass is below 2m(p) at M=1859(+3)(-10) (stat)+5-25(syst) MeV/c(2) and the total width is Gamma<30 MeV/c(2) at the 90% confidence level. These mass and width values are not consistent with the properties of any known particle. PMID- 12906474 TI - Locality of quark-hadron duality and deviations from quark counting rules above the resonance region. AB - We show how deviations from the dimensional scaling laws for exclusive processes may be related to a breakdown in the locality of quark-hadron duality. The essential principles are illustrated in a pedagogic model of a composite system with two spinless charged constituents, for which a dual picture for the low energy resonance phenomena and high-energy scaling behavior can be established. We introduce the concept of "restricted locality" of quark-hadron duality and show how this results in deviations from the perturbative quantum chromodynamics quark counting rules above the resonance region. In particular, it can be a possible source for oscillations about the smooth quark counting rule, as seen, e.g., in the 90-degree differential cross sections for gammap-->pi(+)n. PMID- 12906473 TI - Cross-section measurement of charged-pion photoproduction from hydrogen and deuterium. AB - We have measured the differential cross section for the gamman-->pi(-)p and gammap-->pi(+)n reactions at theta(c.m.)=90 degrees in the photon energy range from 1.1 to 5.5 GeV at Jefferson Lab (JLab). The data at E(gamma) greater, similar 3.3 GeV exhibit a global scaling behavior for both pi(-) and pi(+) photoproduction, consistent with the constituent counting rule and the existing pi(+) photoproduction data. Possible oscillations around the scaling value are suggested by these new data. The data show enhancement in the scaled cross section at a center-of-mass energy near 2.2 GeV. The cross section ratio of exclusive pi(-) to pi(+) photoproduction at high energy is consistent with the prediction based on one-hard-gluon-exchange diagrams. PMID- 12906475 TI - Ion-induced quark-gluon implosion. AB - We investigate nuclear fragmentation in the central proton-nucleus and nucleus nucleus collisions at the energies of CERN LHC. Within the semiclassical approximation we argue that because of the fast increase with energy of the cross sections of soft and hard interactions each nucleon is stripped in the average process off "soft" partons and fragments into a collection of leading quarks and gluons with large p(t). Valence quarks and gluons are streaming in the opposite directions when viewed in the c.m. of the produced system. The resulting pattern of the fragmentation of the colliding nuclei leads to an implosion of the quark and gluon constituents of the nuclei. The nonequilibrium state produced at the initial stage in the nucleus fragmentation region is estimated to have densities >/=50 GeV/fm(3) at the LHC energies and probably >/=10 GeV/fm(3) at BNL RHIC. PMID- 12906472 TI - Measurement of ep-->e' ppi+ pi- and baryon resonance analysis. AB - The cross section for the reaction ep-->e(')ppi(+)pi(-) was measured in the resonance region for 1.4pi+/-pi0, study of B+/--->K+/-pi0, and search for B0-->pi0pi0. AB - We present results for the branching fractions and charge asymmetries in B+/-- >h(+/-)pi(0) (where h(+/-)=pi(+/-),K+/-) and a search for the decay B0- >pi(0)pi(0) using a sample of approximately 88 x 10(6) BBmacr; pairs collected by the BABAR detector at the PEP-II asymmetric-energy B Factory at SLAC. We measure B(B+/--->pi(+/-)pi(0))=(5.5(+1.0)(-0.9)+/-0.6)x10(-6), where the first error is statistical and the second is systematic. The B+/--->pi(+/-)pi(0) signal has a significance of 7.7sigma including systematic uncertainties. We simultaneously measure the K+/-pi(0) branching fraction to be B(B+/--->K+/-pi(0))=(12.8(+1.2)( 1.1)+/-1.0)x10(-6). The charge asymmetries are Api(+/-)(pi(0))=-0.03(+0.18)( 0.17)+/-0.02 and AK+/-(pi(0))=-0.09+/-0.09+/-0.01. We place a 90% confidence level upper limit on the branching fraction B(B0-->pi(0)pi(0)) of 3.6 x 10(-6). PMID- 12906480 TI - Polarization-independent linear waveguides in 3D photonic crystals. AB - Using a symmetry-based approach, we have designed polarization-independent waveguides in a 3D photonic crystal. A comprehensive series of numerical experiments, involving the propagation of pulsed signals through long straight waveguide sections and sharp bends, quantitatively evaluates the bend transmission coefficient over the entire bandwidth of the corresponding guided modes. High (approximately 95%) polarization-independent bend transmission is achieved within a certain frequency range. PMID- 12906481 TI - Three types of transitions to phase synchronization in coupled chaotic oscillators. AB - We study the effect of noncoherence on the onset of phase synchronization of two coupled chaotic oscillators. Depending on the coherence properties of oscillations characterized by the phase diffusion, three types of transitions to phase synchronization are found. For phase-coherent attractors this transition occurs shortly after one of the zero Lyapunov exponents becomes negative. At rather strong phase diffusion, phase locking manifests a strong degree of generalized synchronization, and occurs only after one positive Lyapunov exponent becomes negative. For intermediate phase diffusion, phase synchronization sets in via an interior crises of the hyperchaotic set. PMID- 12906482 TI - Close-packed floating clusters: granular hydrodynamics beyond the freezing point? AB - Monodisperse granular flows often develop regions with hexagonal close packing of particles. We investigate this effect in a system of inelastic hard spheres driven from below by a "thermal" plate. Molecular dynamics simulations show, in a wide range of parameters, a close-packed cluster supported by a low-density region. Surprisingly, the steady-state density profile, including the close packed cluster part, is well described by a variant of Navier-Stokes granular hydrodynamics (NSGH). We suggest a simple explanation for the success of NSGH beyond the freezing point. PMID- 12906483 TI - Generation of a self-chirped few-cycle optical pulse in a FEL oscillator. AB - We study the generation of a self-chirped optical pulse in a free-electron laser (FEL) oscillator. In a high-gain FEL oscillator, the frequency chirp is induced in the slippage region as a result of superradiant FEL resonance, and this time frequency correlation evolves continuously into a few-cycle regime, if the optical cavity length is perfectly synchronized to the electron bunch interval. Numerical simulations based on the slowly evolving wave approximation and experimental results are presented. PMID- 12906484 TI - Continuum self-organized-criticality model of turbulent heat transport in tokamaks. AB - A simple generic one-dimensional continuum model of driven dissipative systems is proposed to explain self-organized bursty heat transport in tokamaks. Extensive numerical simulations of this model reproduce many features of present day tokamaks such as submarginal temperature profiles, intermittent transport events, 1/f scaling of the frequency spectra, propagating fronts, etc. This model utilizes a minimal set of phenomenological parameters, which may be determined from experiments and/or simulations. Analytical and physical understanding of the observed features has also been attempted. PMID- 12906485 TI - Lattice trapping barriers to brittle fracture. AB - We present a multiscale simulation of a crack in silicon under tensile loading that is consistent with experiment; fracture is brittle with a modest lattice trapping energy barrier to crack propagation. Our multiscale molecular-dynamics simulation has a tight-binding description of bonding near the crack tip embedded in an empirical-potential (EP) region. Forces on atoms in the tight-binding region are computed using a Green's function method. Comparing our multiscale simulation with EP simulations shows that the EP models severely overestimate lattice trapping, explaining the failure of the Griffith criterion and the dramatic differences in crack morphology. A two-length-scale model for the lattice-trapping energy barrier correctly predicts the critical load for brittle fracture. We argue that lattice trapping plays an important role in the brittle to-ductile transition. PMID- 12906486 TI - Monolayer ice. AB - We report results from molecular dynamics simulations of water under confinement and at ambient conditions that predict a first-order freezing transition from a monolayer of liquid water to a monolayer of ice induced by increasing the distance between the confining parallel plates. Since a slab geometry is incompatible with a tetrahedral arrangement of the sp(3) hybridized oxygen of water, the freezing is coupled to a linear buckling transition. By exploiting the ordered out-of-plane displacement of the molecules in the buckled phase the distortion of the hydrogen bonds is minimized. PMID- 12906487 TI - Anomalous dislocation multiplication in FCC metals. AB - Direct atomistic simulations of dislocation multiplication in fcc aluminum reveal an unexpected mechanism, in which a Frank-Read source emits dislocations with Burgers vectors different from that of the source itself. The mechanism is traced to a spontaneous nucleation of partial dislocation loops within the stacking fault. Understanding and a quantitative description of this unusual process are achieved through the development of a continuum model for dislocation nucleation based on the coarse-grained dislocation dynamics approach and a minimal amount of atomistic input. PMID- 12906488 TI - First one-dimensional stress-strain experiments inside an aperiodic inclusion compound: evidence of depinning effects. AB - This Letter reports on the first observation of depinning under hydrostatic pressure of the guest and host sublattices inside the aperiodic nonadecane-urea inclusion compound. This leads to a unique way for a direct determination of the one-dimensional interguest interaction (1 N/m for nonadecane). PMID- 12906489 TI - Metastable Frenkel pair defect in graphite: source of Wigner energy? AB - The atomic processes associated with energy storage and release in irradiated graphite have long been subject to untested speculation. We examine structures and recombination routes for interstitial-vacancy (I-V) pairs in graphite. Interaction results in the formation of a new metastable defect (an intimate I-V pair) or a Stone-Wales defect. The intimate I-V pair, although 2.9 eV more stable than its isolated constituents, still has a formation energy of 10.8 eV. The barrier to recombination to perfect graphite is calculated to be 1.3 eV, consistent with the experimental first Wigner energy release peak at 1.38 eV. We expect similar defects to form in carbon nanostructures such as nanotubes, nested fullerenes, and onions under irradiation. PMID- 12906490 TI - New mechanism for the alpha to omega martensitic transformation in pure titanium. AB - We propose a new direct mechanism for the pressure driven alpha-->omega martensitic transformation in pure titanium. A systematic algorithm enumerates all possible pathways whose energy barriers are evaluated. A new, homogeneous pathway emerges with a barrier at least 4 times lower than other pathways. The pathway is shown to be favorable in any nucleation model. PMID- 12906491 TI - Critical behavior of thermal relaxation near a breakdown point. AB - At a composition far above the percolation threshold, the resistance of a composite sample increases with time due to Joule heating as a constant current of a sufficiently large value is passed through the sample. If the current is less than a certain breakdown current (I(b)) the resistance eventually reaches a steady value with a characteristic relaxation time tau(h). The latter diverges with current I as tau(h) approximately (1-I(2)/I(2)(b))(-z). The value of the exponent z displays large fluctuations leading to unusual scaling of the relaxation time. It is shown that the results lead to important conclusions about the nature of breakdown phenomena. PMID- 12906492 TI - Evidence of 1D behavior of He4 confined within carbon-nanotube bundles. AB - We present the first low-temperature thermodynamic investigation of the controlled physisorption of He4 gas in carbon single-wall nanotube (SWNT) samples. The vibrational specific heat measured between 100 mK and 6 K demonstrates an extreme sensitivity to outgassing conditions. For bundles with a few number of nanotubes the extra contribution to the specific heat, C(ads), originating from adsorbed He4 at very low density displays 1D behavior, typical for He atoms localized within linear channels as grooves and interstitials, for the first time evidenced. For larger bundles, C(ads) recovers the 2D behavior akin to the case of He4 films on planar substrates (grafoil). PMID- 12906493 TI - Grains, growth, and grooving. AB - We report the in situ investigation of grain growth and grain boundary migration, performed with a variable-temperature scanning tunneling microscope (STM) on a polycrystalline gold film. Atomic step resolution allowed us to identify the individual grains and, thus, also the grain boundaries. Our special, thermal drift-compensated STM design made it possible to follow the same sample area over large temperature intervals. In this way, we have directly observed grain boundary migration and grain growth. In a first quantitative analysis we correlate the observed, unexpected changes in surface roughness with the evolution of the grain and grain boundary configuration. PMID- 12906494 TI - Knudsen diffusivity of a hard sphere in a rough slit pore. AB - An analytic theory for the Knudsen self-diffusivity D(s) of hard spheres in an atomically rough slit-shaped pore is presented which quantitatively matches simulation results. The theory assumes that, due to chaotic molecular trajectories caused by surface morphology, collisions of gas molecules with the wall are partly diffuse and partly specular, the relative magnitude of each depending upon the magnitude of the tangential momentum accommodation coefficient f. The theory thus represents a universal Knudsen fluctuation-dissipation correlation between longitudinal momentum loss and diffusivity that can simplify efforts to estimate D(s). It is also found that D(s) computed using Maxwell's theory of slip, in which collisions with the walls are assumed to be purely diffuse or specular, overpredicts the simulated D(s) by a large margin. PMID- 12906495 TI - Nucleation and growth of molecular organic crystals in a liquid film under vapor deposition. AB - We report on the nucleation and growth of tetracene in a thin liquid film which is continuously supersaturated by vapor deposition of molecules onto the film. In a first stage, nucleation and fast anisotropic two-dimensional dendritic growth occurs. In a second stage, the dendrites coarsen into pallet-shaped crystals. These are highly oriented with respect to the plane of the liquid film and reach a lateral size of several 100 microm. The two-dimensional growth mode is explained by the confined growth geometry in the liquid in combination with the anisotropy of the crystal structure. PMID- 12906496 TI - Doping of magic nanoclusters in the submonolayer In/Si(100) system. AB - Si(100)4 x 3-In reconstruction is essentially a superlattice of magic (identical size) Si7In6 nanoclusters. Using scanning tunneling microscopy (STM) observations, we have found that under appropriate growth conditions up to 35% of these clusters can be modified; namely, two Si atoms in the cluster can be replaced by two In atoms, thus forming a Si5In8 cluster. This modification can be considered as a doping of the magic cluster, as it changes the electronic properties of the cluster from semiconducting towards metallic. The doped cluster is less rigid than the ordinary one and swings in the electrical field of the STM tip. The atomic structure and stability of the doped magic cluster have been examined using first-principles total-energy calculations. PMID- 12906497 TI - Locally critical point in an anisotropic Kondo lattice. AB - We report the first numerical identification of a locally quantum critical point at which the criticality of the local Kondo physics is embedded in that associated with a magnetic ordering. We are able to numerically access the quantum critical behavior by focusing on a Kondo-lattice model with Ising anisotropy. We also establish that the critical exponent for the q-dependent dynamical spin susceptibility is fractional and compares well with the experimental value for heavy fermions. PMID- 12906498 TI - How irrelevant operators affect the determination of fractional charge. AB - We show that the inclusion of irrelevant terms in the Hamiltonian describing tunneling between edge states in the fractional quantum Hall effect can lead to a variety of nonperturbative behaviors in intermediate energy regimes and, in particular, affect crucially the determination of charge through shot noise measurements. We show, for instance, that certain combinations of relevant and irrelevant terms can lead to an effective measured charge nue in the strong backscattering limit and an effective measured charge e in the weak backscattering limit, in sharp contrast with standard perturbative expectations. This provides a possible scenario to explain the experimental observations by Comforti et al. [Nature (London) 416, 515 (2002)]], which are so far not understood. PMID- 12906499 TI - Ionic shell and subshell structures in aluminum and gold nanocontacts. AB - Conductance histograms of aluminum and gold nanocontact rupture are studied experimentally and simulated using embedded atom potentials to assess the interplay between electronic and structural properties at room temperature. Our results reveal a crossover from quantized conductance structures to crystalline faceting or geometric shell/subshell structures at 300 K. The absence of electronic shell structure in gold and aluminum is in stark contrast with the behavior of alkaline metal nanowires which emulate their cluster counterparts. Semiclassical arguments suggest why rapid dominance of ionic structures takes place, and possible nanowire architectures are proposed in consistency with both the experimental and simulated nanocontact data. PMID- 12906500 TI - Spin-blockade spectroscopy of a two-level artificial molecule. AB - Coulomb- and spin-blockade spectroscopy investigations have been performed on an electrostatically defined "artificial molecule" connected to spin polarized leads. The molecule is first effectively reduced to a two-level system by placing both constituent atoms at a specific location of the level spectrum. The spin sensitivity of the conductance enables us to identify the electronic spin states of the two-level molecule. We find in addition that the magnetic field induces variations in the tunnel coupling between the two atoms. The lateral nature of the device is evoked to explain this behavior. PMID- 12906501 TI - Even-odd effect in spontaneously coherent bilayer quantum Hall droplets. AB - Using exact diagonalization in the disk geometry we predict a novel even-odd effect in the Coulomb-blockade spectra of vertically coupled double quantum dots under an external magnetic field. The even-odd effect in the tunneling conductance is a direct manifestation of spontaneous interlayer phase coherence, and is similar to the even-odd resonance in the Cooper pair box problem in mesoscopic superconducting grains. Coherent fluctuations in the number of Cooper pairs in superconductors are analogous to the fluctuations in the relative number difference between the two layers in quantum Hall droplets. PMID- 12906502 TI - Metallic behavior of lightly doped La2-xSrxCuO4 with a Fermi surface forming an arc. AB - Lightly doped La2-xSrxCuO4 in the so-called "insulating" spin-glass phase has been studied by angle-resolved photoemission spectroscopy. We have observed that a "quasiparticle" (QP) peak crosses the Fermi level in the node direction of the d-wave superconducting gap, forming an "arc" of Fermi surface, which explains the metallic behavior at high temperatures of the lightly doped materials. The QP spectral weight of the arc smoothly increases with hole doping, which we attribute to the n approximately x behavior of the carrier number in the underdoped and lightly doped regions. PMID- 12906503 TI - Disordered nanocrystalline superconducting PbMo6S8 with a very large upper critical field. AB - Large increases in the upper critical field B(C2)(0) are reported in bulk superconductors that demonstrate another novel property for nanocrystalline materials. Disordered nanocrystalline PbMo6S8 superconductors were fabricated by mechanical milling and hot isostatic pressing. Conventional PbMo6S8 has B(C2)(0) approximately 50 T. The nanocrystalline materials have higher resistivity (rho(N)) and B(C2)(0) approximately 100 T. The disorder produced in these nanocrystalline materials is significantly different from that produced by doping because it increases rho(N) and, hence, B(C2)(0) without significantly reducing the electronic density of states or superconducting transition temperature (T(C)). Furthermore, the disorder reduces the electron mean-free path to approximately 1 nm which is more than an order of magnitude smaller than the grain size and necessary to achieve the unprecedented increase in B(C2)(0). PMID- 12906504 TI - Perpendicular interlayer coupling in Ni80Fe20/NiO/Co trilayers. AB - An in-plane perpendicular magnetic coupling between Ni80Fe20 and Co has been found in NiFe/NiO/Co trilayers for a NiO thickness ranging from 4 to 25 nm by magneto-optical Kerr effect and x-ray magnetic circular dichroism measurements. In the easy magnetization direction of the Co layer, the Co coercive field H(C) increases when the thickness of the NiO layer t(NiO) increases. Because of the coupling, H(C) is always larger than for NiO/Co bilayers with the same thicknesses. The saturation field of the NiFe layer H(S) decreases when t(NiO) increases, indicating a weakening of the coupling. Numerical simulations show that the presence of interface roughness combined with a small value of the NiO anisotropy can explain the observed 90 degrees coupling. PMID- 12906505 TI - Local magnetic susceptibility of the positive muon in the quasi-one-dimensional S=1/2 antiferromagnet dichlorobis (pyridine) copper (II). AB - We report muon spin rotation measurements of the local magnetic susceptibility around a positive muon in the paramagnetic state of the quasi-one-dimensional spin 1/2 antiferromagnet dichlorobis (pyridine) copper (II). Signals from three distinct sites are resolved and have a temperature dependent frequency shift which is significantly different than the magnetic susceptibility. This difference is attributed to a muon induced perturbation of the spin 1/2 chain. The obtained frequency shifts are compared with transfer matrix density-matrix renormalization-group numerical simulations. PMID- 12906506 TI - Exchange interaction effects on the optical properties of LuMnO3. AB - We have measured the optical conductivity of single crystal LuMnO3 from 10 to 45000 cm(-1) at temperatures between 4 and 300 K. A symmetry allowed on-site Mn d d transition near 1.7 eV is observed to blueshift ( approximately 0.1 eV) in the antiferromagnetic state due to Mn-Mn superexchange interactions. Similar anomalies are observed in the temperature dependence of the TO phonon frequencies which arise from spin-phonon interaction. We find that the known anomaly in the temperature dependence of the quasistatic dielectric constant epsilon(0) below T(N) approximately 90 K is overwhelmingly dominated by the phonon contributions. PMID- 12906507 TI - Quasiparticle relaxation dynamics in heavy fermion compounds. AB - We present the first femtosecond studies of electron-phonon (e-ph) thermalization in heavy-fermion compounds. The e-ph thermalization time tau(ep) increases below the Kondo temperature by more than 2 orders of magnitude as T=0 K is approached. Analysis using the two-temperature model and numerical simulations based on Boltzmann's equations suggest that this anomalous slowing down of the e-ph thermalization derives from the large electronic specific heat and the suppression of scattering between heavy electrons and phonons. PMID- 12906508 TI - Classical and quantum communication without a shared reference frame. AB - We show that communication without a shared reference frame is possible using entangled states. Both classical and quantum information can be communicated with perfect fidelity without a shared reference frame at a rate that asymptotically approaches one classical bit or one encoded qubit per transmitted qubit. We present an optical scheme to communicate classical bits without a shared reference frame using entangled photon pairs and linear optical Bell state measurements. PMID- 12906509 TI - Programmable networks for quantum algorithms. AB - The implementation of a quantum computer requires the realization of a large number of N-qubit unitary operations which represent the possible oracles or which are part of the quantum algorithm. Until now there have been no standard ways to uniformly generate whole classes of N-qubit gates. We develop a method to generate arbitrary controlled phase-shift operations with a single network of one qubit and two-qubit operations. This kind of network can be adapted to various physical implementations of quantum computing and is suitable to realize the Deutsch-Jozsa algorithm as well as Grover's search algorithm. PMID- 12906510 TI - Exact two-qubit universal quantum circuit. AB - We provide an analytic way to implement any arbitrary two-qubit unitary operation, given an entangling two-qubit gate together with local gates. This is shown to provide explicit construction of a universal quantum circuit that exactly simulates arbitrary two-qubit operations in SU(4). Each block in this circuit is given in a closed form solution. We also provide a uniform upper bound of the applications of the given entangling gates, and find that exactly half of all the controlled-unitary gates satisfy the same upper bound as the CNOT gate. These results allow for the efficient implementation of operations in SU(4) required for both quantum computation and quantum simulation. PMID- 12906511 TI - Discontinuous unbinding of lipid multibilayers. AB - We have observed a discontinuous unbinding transition of lipid bilayer stacks composed of phosphatidylethanolamine and phosphatidylglycerol using x-ray diffraction. The unbinding is reversible and coincides with the main (L(beta)- >L(alpha)) transition of the lipid mixture. Interbilayer interaction potentials deduced from the diffraction data reveal that the bilayers in the L(beta) phase are only weakly bound. The unbinding transition appears to be driven by an abrupt increase in steric repulsion resulting from increased thermal undulations of the bilayers upon entering the fluid L(alpha) phase. PMID- 12906512 TI - Saltatory waves in the spike-diffuse-spike model of active dendritic spines. AB - In this Letter we present the explicit construction of a saltatory traveling pulse of nonconstant profile in an idealized model of dendritic tissue. Excitable dendritic spine clusters, modeled with integrate-and-fire (IF) units, are connected to a passive dendritic cable at a discrete set of points. The saltatory nature of the wave is directly attributed to the breaking of translation symmetry in the cable. The conditions for propagation failure are presented as a function of cluster separation and IF threshold. PMID- 12906513 TI - Dynamic force spectroscopy of protein-DNA interactions by unzipping DNA. AB - We demonstrate the first site-specific single-molecule characterization of the prominent activation barrier for the disruption of a protein-DNA binding complex. We achieved this new capability by combining dynamic force spectroscopy with unzipping force analysis of protein association and used the combination to investigate restriction enzyme binding to specific DNA sites. Analysis revealed lifetimes and interaction distances for three protein-DNA interactions. This new method is able to distinguish protein-DNA binding complexes on a site-specific, single-molecule basis. PMID- 12906514 TI - Ion multivalence and like-charge polyelectrolyte attraction. AB - It is known empirically that multivalent ions generate attractions between like charged polyelectrolytes, with different valence requirements for different systems. How multivalent must an ion be before it can condense a given polyelectrolyte? Using charge-tunable M13 virus rods and a family of artificial homologous "dumbbell" divalent ions of different sizes, we have constructed a multivalent ion-polyelectrolyte phase diagram, and find an experimentally motivated general criterion for like-charged attraction based on the ion valence, ion size, and the Gouy-Chapman length. PMID- 12906515 TI - Does a single zealot affect an infinite group of voters? AB - A method for studying the exact properties of a class of inhomogeneous stochastic many-body systems is developed and presented in the framework of a voter model perturbed by the presence of a "zealot," an individual allowed to favor an "opinion." We compute exactly the magnetization of this model and find that in one (1D) and two dimensions (2D) it evolves, algebraically ( approximately t( 1/2)) in 1D and much slower ( approximately 1/ln(t) in 2D, towards the unanimity state chosen by the zealot. In higher dimensions the stationary magnetization is no longer uniform: the zealot cannot influence all the individuals. The implications to other physical problems are also pointed out. PMID- 12906516 TI - Comment on "Repulsive Casimir forces". PMID- 12906518 TI - Comment on "Transverse NMR relaxation as a probe of mesoscopic structure". PMID- 12906520 TI - Fresnel representation of the Wigner function: an operational approach. AB - We present an operational definition of the Wigner function. Our method relies on the Fresnel transform of measured Rabi oscillations and applies to motional states of trapped atoms as well as to field states in cavities. We illustrate this technique using data from recent experiments in ion traps [Phys. Rev. Lett. 76, 1796 (1996)]] and in cavity QED [Nature (London) 403, 743 (2000)]]. The values of the Wigner functions of the underlying states at the origin of phase space are W(|0>)(0)=+1.75 for the vibrational ground state and W(|1>)(0)=-1.4 for the one-photon number state. We generalize this method to wave packets in arbitrary potentials. PMID- 12906521 TI - Entanglement generation and multiparticle interferometry with neutral atoms. AB - We study the preparation and manipulation of states involving a small number of interacting particles. By controlling the splitting and fusing of potential wells, we show how to interconvert Mott-insulator-like and trapped BEC-like states. We also discuss the generation of "Schrodinger cat" states by splitting a microtrap and taking into practical consideration the asymmetry between the resulting wells. These schemes can be used to perform multiparticle interferometry with neutral atoms, where interference effects can be observed only when all the participating particles are measured. PMID- 12906522 TI - Energetically stable particlelike skyrmions in a trapped Bose-Einstein condensate. AB - We numerically show that a topologically nontrivial 3D Skyrmion can be energetically stable in a trapped two-component atomic Bose-Einstein condensate, for the parameters of 87Rb condensate experiments. The separate conservation of the two atomic species can stabilize the Skyrmion against shrinking to zero size, while drift of the Skyrmion due to the trap-induced density gradient can be prevented by rotation or by a laser potential. PMID- 12906523 TI - Quantum nonlocality in the presence of superselection rules and data hiding protocols. AB - We consider a quantum system subject to superselection rules, for which certain restrictions apply to the quantum operations that can be implemented. It is shown how the notion of quantum nonlocality has to be redefined in the presence of superselection rules: there exist separable states that cannot be prepared locally and exhibit some form of nonlocality. Moreover, the notion of local distinguishability in the presence of classical communication has to be altered. This can be used to perform quantum information tasks that are otherwise impossible. In particular, this leads to the introduction of perfect quantum data hiding protocols, for which quantum communication (eventually in the form of a separable but nonlocal state) is needed to unlock the secret. PMID- 12906524 TI - Momentum spectroscopy of 1D phase fluctuations in Bose-Einstein condensates. AB - We measure the axial momentum distribution of Bose-Einstein condensates with an aspect ratio of 152 using Bragg spectroscopy. We observe the Lorentzian momentum distribution characteristic of one-dimensional phase fluctuations. The temperature dependence of the width of this distribution provides a quantitative test of quasicondensate theory. In addition, we observe a condensate length consistent with the suppression of density fluctuations, even when phase fluctuations are large. PMID- 12906525 TI - Measurement of the spatial correlation function of phase fluctuating Bose Einstein condensates. AB - We measure the intensity correlation function of two interfering spatially displaced copies of phase fluctuating Bose-Einstein condensates. It is shown that this corresponds to a measurement of the phase correlation properties of the initial condensate. Analogous to the method used in the stellar interferometer experiment of Hanbury Brown and Twiss, we use spatial intensity correlations to determine the phase coherence lengths of elongated condensates. We find good agreement with our prediction of the correlation function and confirm the expected coherence length. PMID- 12906526 TI - Coherent transport of neutral atoms in spin-dependent optical lattice potentials. AB - We demonstrate the controlled coherent transport and splitting of atomic wave packets in spin-dependent optical lattice potentials. Such experiments open intriguing possibilities for quantum state engineering of many body states. After first preparing localized atomic wave functions in an optical lattice through a Mott insulating phase, we place each atom in a superposition of two internal spin states. Then state selective optical potentials are used to split the wave function of a single atom and transport the corresponding wave packets in two opposite directions. Coherence between the wave packets of an atom delocalized over up to seven lattice sites is demonstrated. PMID- 12906527 TI - Controlling transport in mixtures of interacting particles using Brownian motors. AB - An outstanding open problem in nanoscience is how to control the motion of tiny particles. Ratchetlike devices, inspired by biological motors, have been proposed as a way to achieve this goal. However, the net directed transport is almost suppressed if the diffusing particles are weakly coupled to the underlying spatially asymmetric substrate. Here we show how adding particles of an auxiliary species, that interact with both the primary particles of interest and the substrate, provides a controlled enhancement of the flow for both species. These can move either together or in opposite directions, depending upon the strength of the interaction, and whether it is attractive or repulsive. PMID- 12906528 TI - Fractional diffusion in the multiple-trapping regime and revision of the equivalence with the continuous-time random walk. AB - We investigate the macroscopic diffusion of carriers in the multiple-trapping (MT) regime, in relation with electron transport in nanoscaled heterogeneous systems, and we describe the differences, as well as the similarities, between MT and the continuous-time random walk (CTRW). Diffusion of free carriers in MT can be expressed as a generalized continuity equation based on fractional time derivatives, while the CTRW model for diffusive transport generalizes the constitutive equation for the carrier flux. PMID- 12906529 TI - Methods for constraining fine structure constant evolution with OH microwave transitions. AB - We investigate the constraints that OH microwave transitions in megamasers and molecular absorbers at cosmological distances may place on the evolution of the fine structure constant alpha=e(2)/ variant Planck's over 2pi c. The centimeter OH transitions are a combination of hyperfine splitting and lambda doubling that can constrain the cosmic evolution of alpha from a single species, avoiding systematic errors in alpha measurements from multiple species which may have relative velocity offsets. The most promising method compares the 18 and 6 cm OH lines, includes a calibration of systematic errors, and offers multiple determinations of alpha in a single object. Comparisons of OH lines to the HI 21 cm line and CO rotational transitions also show promise. PMID- 12906530 TI - Superweakly interacting massive particles. AB - We investigate a new class of dark matter: superweakly interacting massive particles (super-WIMPs). As with conventional WIMPs, super-WIMPs appear in well motivated particle theories with naturally the correct relic density. In contrast to WIMPs, however, super-WIMPs are impossible to detect in all conventional dark matter searches. We consider the concrete examples of gravitino and graviton cold dark matter in models with supersymmetry and universal extra dimensions, respectively, and show that super-WIMP dark matter satisfies stringent constraints from big bang nucleosynthesis and the cosmic microwave background. PMID- 12906531 TI - Left-right model of Quark and Lepton masses without a scalar bidoublet. AB - We propose a left-right model of quarks and leptons based on the gauge group SU(3)(C)xSU(2)(L)xSU(2)(R)xU(1)(B-L), where the scalar sector consists of only two doublets: (1,2,1,1) and (1,1,2,1). As a result, any fermion mass, whether it be Majorana or Dirac, must come from dimension-five operators. This allows us to have a common view of quark and lepton masses, including the smallness of Majorana neutrino masses as the consequence of a double seesaw mechanism. PMID- 12906533 TI - New relations for excited baryons in large-N(c) QCD. AB - We show that excited baryons in large-N(c) QCD form multiplets, within which masses are first split at O(1/N(c)). The dominant couplings of resonances to various mesons are highly constrained: The N(1535) decays at leading 1/N(c) order exclusively to eta-N rather than pi-N, and vice versa for the N(1650). This multiplet structure is reproduced by a simple large-N(c) quark model, well studied in the literature, that describes resonances as single-quark excitations. PMID- 12906534 TI - Evidence for a narrow S = +1 baryon resonance in photoproduction from the neutron. AB - The gamman-->K(+)K(-)n reaction on 12C has been studied by measuring both K+ and K- at forward angles. A sharp baryon resonance peak was observed at 1.54+/-0.01 GeV/c(2) with a width smaller than 25 MeV/c(2) and a Gaussian significance of 4.6sigma. The strangeness quantum number (S) of the baryon resonance is +1. It can be interpreted as a molecular meson-baryon resonance or alternatively as an exotic five-quark state (uuddsmacr;) that decays into a K+ and a neutron. The resonance is consistent with the lowest member of an antidecuplet of baryons predicted by the chiral soliton model. PMID- 12906535 TI - Observed Ds(2317) and tentative D(2100-2300) as the charmed cousins of the light scalar nonet. AB - The very recently observed D(*)(sJ)(2317)(+) meson is described as a quasibound scalar csmacr; state in a unitarized meson model, owing its existence to the strong 3P0 Okubo-Zweig-Iizuka-allowed coupling to the nearby S-wave DK threshold. By the same mechanism, a scalar D(*)(0)(2100-2300) resonance is predicted above the Dpi threshold. These scalars are the charmed cousins of the light scalar nonet f(0)(600), f(0)(980), K(*)(0)(800), and a(0)(980), reproduced by the same model. The standard cnmacr; and csmacr; charmed scalars D0 and D(s0), cousins of the scalar nonet f(0)(1370), f(0)(1500), K(*)(0)(1430), and a(0)(1450), are predicted to lie at about 2.64 and 2.79 GeV, respectively, both with a width of some 200 MeV. PMID- 12906536 TI - New direct reaction: two-proton knockout from neutron-rich nuclei. AB - The reaction 9Be(28Mg,26Ne+gamma)X has been studied at 82 MeV/nucleon together with two similar cases, 30Mg and 34Si. Strong evidence that the reactions are direct is offered by the parallel-momentum distributions of the reaction residues and by the inclusive cross sections. The pattern of the partial cross sections for 28Mg suggests the presence of correlations. A preliminary theoretical discussion based on eikonal reaction theory and the many-body shell model is presented. The reaction holds great promise for the study of neutron-rich nuclei. PMID- 12906537 TI - Neutrino-12C scattering in the Ab initio shell model with a realistic three-body interaction. AB - We investigate cross sections for neutrino-12C exclusive scattering and for muon capture on 12C using wave functions obtained in the ab initio no-core shell model. In our parameter-free calculations with basis spaces up to the 6 variant Planck's over 2pi Omega we show that realistic nucleon-nucleon interactions, like, e.g., the CD-Bonn, underpredict the experimental cross sections by more than a factor of 2. By including a realistic three-body interaction, Tucson Melbourne TM'(99), the cross sections are enhanced significantly and a much better agreement with experiment is achieved. At the same time, the TM'(99) interaction improves the calculated level ordering in 12C. The comparison between the CD-Bonn and the three-body calculations provides strong confirmation for the need to include a realistic three-body interaction to account for the spin-orbit strength in p-shell nuclei. PMID- 12906538 TI - Differential cross sections from quantum calculations on coupled Ab initio potential energy surfaces and scattering experiments for Cl(2P)+H2 reactions. AB - To assess the relative reactivity of the spin-orbit excited state of atomic Cl with molecular hydrogen, we have measured differential cross sections using an atomic Cl beam with a known concentration of the ground and excited spin-orbit states. These are compared with the first determination of the cross sections from quantum mechanical scattering calculations on a set of coupled ab initio potential energy surfaces. The comparison suggests that these surfaces may underestimate the degree of rotational excitation of the HCl products and that the excited spin-orbit state plays a minor role in the reaction. PMID- 12906539 TI - Heterogeneity in oscillator networks: are smaller worlds easier to synchronize? AB - Small-world and scale-free networks are known to be more easily synchronized than regular lattices, which is usually attributed to the smaller network distance between oscillators. Surprisingly, we find that networks with a homogeneous distribution of connectivity are more synchronizable than heterogeneous ones, even though the average network distance is larger. We present numerical computations and analytical estimates on synchronizability of the network in terms of its heterogeneity parameters. Our results suggest that some degree of homogeneity is expected in naturally evolved structures, such as neural networks, where synchronizability is desirable. PMID- 12906540 TI - Fluctuating particle motion during shear induced granular compaction. AB - Using a refractive index matching method, we investigate the trajectories of particles in three dimensional granular packing submitted to cyclic shear deformation. The particle motion observed during compaction is not diffusive but exhibits a transient cage effect, similar to the one observed in colloidal glasses. We precisely study the statistics of the step size between two successive cycles and observe that it is proportional to the shear amplitude. The link between the microscopic observations and the macroscopic evolution of the volume fraction during compaction is discussed. PMID- 12906541 TI - Effects of air on the segregation of particles in a shaken granular bed. AB - Effects of interstitial air on the motions of a large intruder in a shaken granular bed are studied experimentally as a function of ambient air pressure, particle size of the bed, and the density of the intruder. It is found that the intruder always rises from the granular bed in the absence of air. However, the intruder can acquire both positive and negative buoyancy in the presence of air. Negative buoyancy can be observed only when both the density of the intruder and the particle size of the bed are small enough. This negative buoyancy can be explained by the unusual air pressure distribution found in the bed. PMID- 12906542 TI - Sandpile formation by revolving rivers. AB - Experimental observation of a new mechanism of sandpile formation is reported. As a steady stream of dry sand is poured onto a horizontal surface, a pile forms which has a thin river of sand on one side flowing from the apex of the pile to the edge of its base. The river rotates about the pile, depositing a new layer of sand with each revolution, thereby causing the pile to grow. For small piles the river is steady and the pile formed is smooth. For larger piles, the river becomes intermittent and the surface of the pile becomes undulating. The essential features of the system that produce the phenomenon are discussed, and the robustness of the phenomena is demonstrated with experiments using different boundary conditions and sands. PMID- 12906532 TI - Central pseudorapidity gaps in events with a leading antiproton at the fermilab tevatron pp collider. AB - We report a measurement of the fraction of events with a large pseudorapidity gap deltaeta within the pseudorapidity region available to the proton dissociation products X in p+p-->p+X. For a final state p of fractional momentum loss xi(p) and 4-momentum transfer squared t(p) within 0.063 is found to be 0.246+/-0.001 (stat)+/-0.042 (syst) [0.184+/-0.001 (stat)+/-0.043 (syst)]. Our results are compared with gap fractions measured in minimum bias pp collisions and with theoretical expectations. PMID- 12906543 TI - Is the electron radiation length constant at high energies? AB - Experimental results for the radiative energy loss of 149, 207, and 287 GeV electrons in a thin Ir target are presented. From the data we conclude that at high energies the radiation length increases in accordance with the Landau Pomeranchuk-Migdal (LPM) theory and thus electrons become more penetrating the higher the energy. The increase of the radiation length as a result of the LPM effect has a significant impact on the behavior of high-energy electromagnetic showers. PMID- 12906544 TI - Cohesive acceleration and focusing of relativistic electrons in overdense plasma. AB - We describe our studies of the generation of plasma wake fields by a relativistic electron bunch and of phasing between the longitudinal and transverse fields in the wake. The leading edge of the electron bunch excites a high-amplitude plasma wake inside the overdense plasma column, and the acceleration and focusing wake fields are probed by the bunch tail. By monitoring the dependence of the acceleration upon the plasma's density, we approached the beam-matching condition and achieved an energy gain of 0.6 MeV over the 17 mm plasma length, corresponding to an average acceleration gradient of 35 MeV/m. Wake-induced modulation in energy and angular divergence of the electron bunch are mapped within a wide range of plasma density. We confirm a theoretical prediction about the phase offset between the accelerating and focusing components of plasma wake. PMID- 12906545 TI - High-intensity laser induced ion acceleration from heavy-water droplets. AB - Fusion neutrons from a heavy water droplet target irradiated with laser pulses of 3 x 10(19) W/cm(2) and from a deuterated secondary target are observed by a time of-flight (TOF) neutron spectrometer. The observed TOF spectrum can be explained by fusion of deuterium ions simultaneously originating from two different sources: ion acceleration in the laser focus by ponderomotively induced charge separation and target-normal sheath acceleration off the target rear surface. The experimental findings agree well with 3D particle-in-cell simulations. PMID- 12906546 TI - Inert gas clusters ejected from bursting bubbles during sputtering. AB - Ar(+)(n) cluster ions (n/=5 Pt layers. Electronic structure calculations confirm the experimental observations, in particular, the dramatic decrease of the CO adsorption energy on a single Pt layer which is caused by the strong Pt-Ru interlayer coupling. PMID- 12906556 TI - Nanocrystal formation and faceting instability in Al(110) homoepitaxy: true upward adatom diffusion at step edges and island corners. AB - Using atomic force microscopy and spot-profile analyzing low energy electron diffraction, we have observed the existence of a striking faceting instability in Al(110) homoepitaxy, characterized by the formation of nanocrystals with well defined facets. These hut-shaped nanocrystals are over tenfold higher than the total film coverage, and coexist in a bimodal growth mode with much shallower and more populous surface mounds. We further use density functional theory calculations to elucidate the microscopic origin of the faceting instability, induced by surprisingly low activation barriers for adatom ascent at step edges and island corners. PMID- 12906557 TI - Phonon softening, chaotic motion, and order-disorder transition in Sn/Ge(111). AB - The phonon dynamics of the Sn/Ge(111) interface is studied using high-resolution helium atom scattering and first-principles calculations. At room temperature we observe a phonon softening at the Kmacr; point in the (sqrt[3]xsqrt[3])R30 degrees phase, associated with the stabilization of a (3x3) phase at low temperature. That phonon band is split into three branches in the (3x3) phase. We analyze the character of these phonons and find out that the low- and room temperature modes are connected via a chaotic motion of the Sn atoms. The system is shown to present an order-disorder transition. PMID- 12906558 TI - Direct imaging of nanoparticle embedding to probe viscoelasticity of polymer surfaces. AB - Atomic force microscopy was used to study the embedding of gold nanoparticles into the surface of polystyrene films. The rate of embedding was determined at temperatures near the bulk glass transition temperature T(g) by measuring the apparent nanosphere height as a function of annealing time. In particular, relative height measurements of the adhered particles were made at temperatures below the bulk T(g) value. In the absence of enhanced surface dynamics or yield processes no embedding is expected to occur for T0.5) inputs, the optimal control is smooth and unique. The optimal variance obtained in the current paper sets the lowest possible bound in controlling the stochasticity of neuronal activity. We also discuss how to implement the optimal control signal for certain model neurons. PMID- 12906578 TI - Adherence of packing defects in soluble proteins. AB - For protein structure to prevail in water, its backbone hydrogen bonds must be shielded from water attack, requiring a cluster of "wrapping" nonpolar groups. Thus, underwrapped regions are adhesive, as exogenous removal of surrounding water becomes thermodynamically favorable. Here we measure the average adhesive force exerted by an underwrapped hydrogen bond on a test hydrophobe and thus define a new interactivity constant. PMID- 12906579 TI - Lamellar phase of stacked two-dimensional rafts of actin filaments. AB - We examined liquid crystalline phases of the cytoskeletal polyelectrolyte filamentous (F-)actin in the presence of multivalent counterions. As a function of increasing ion concentration, the F-actin rods in either an isotropic or a nematic phase will transform into a new and unexpected lamellar phase of cross linked rafts (L(XR) phase), before condensing into a bundled phase of parallel, close-packed rods. This behavior is generic for alkali earth divalent ions Mg2+, Ca2+, Sr2+, and Ba2+, and the structural transitions are achieved without any architecture-specific actin-binding linker proteins. PMID- 12906580 TI - Growth of walled cells: from shells to vesicles. AB - The growth of isolated walled cells is investigated. Examples of such cells range from bacteria to giant algae, and include cochlear hair, plant root hair, fungi, and yeast cells. They are modeled as elastic shells containing a liquid. Cell growth is driven by fluid pressure and is is similar to a plastic deformation of the wall. The requirement of mechanical equilibrium leads to two new scaling laws for cell size that are in quantitative agreement with the compiled biological data. Given these results, possible shapes for growing cells are computed by analogy with those of vesicle membranes. PMID- 12906581 TI - Monte Carlo simulation of a coarse-grained model of polyelectrolyte networks. AB - The structure and properties of a coarse-grained model of a polyelectrolyte network is studied by means of Monte Carlo simulations. Counterions are treated explicitly, and permanent tetrafunctional cross-linking sites are annealed. The resulting pressure-density relationships exhibit a strong dependence on the strength of electrostatic interactions. A discontinuous volume change is observed when electrostatic interactions are strong. The structure of the model networks is examined at various conditions, and it is found to be considerably different from that of noncross-linked polyelectrolytes. PMID- 12906582 TI - Front dynamics in reaction-diffusion systems with Levy flights: a fractional diffusion approach. AB - The use of reaction-diffusion models rests on the key assumption that the diffusive process is Gaussian. However, a growing number of studies have pointed out the presence of anomalous diffusion, and there is a need to understand reactive systems in the presence of this type of non-Gaussian diffusion. Here we study front dynamics in reaction-diffusion systems where anomalous diffusion is due to asymmetric Levy flights. Our approach consists of replacing the Laplacian diffusion operator by a fractional diffusion operator of order alpha, whose fundamental solutions are Levy alpha-stable distributions that exhibit power law decay, x(-(1+alpha)). Numerical simulations of the fractional Fisher-Kolmogorov equation and analytical arguments show that anomalous diffusion leads to the exponential acceleration of the front and a universal power law decay, x(-alpha), of the front's tail. PMID- 12906583 TI - Superfluid Fermi gases with large scattering length. AB - We report quantum Monte Carlo calculations of superfluid Fermi gases with short range two-body attractive interactions with infinite scattering length. The energy of such gases is estimated to be (0.44+/-0.01) times that of the noninteracting gas, and their pairing gap is approximately twice the energy per particle. PMID- 12906584 TI - Measurement of the Casimir force between dissimilar metals. AB - The first precise measurement of the Casimir force between dissimilar metals is reported. The attractive force, between a Cu layer evaporated on a microelectromechanical torsional oscillator and an Au layer deposited on an Al2O3 sphere, was measured dynamically with a noise level of 6 fN/sqrt[Hz]. Measurements were performed for separations in the 0.2-2 micro m range. The results agree to better than 1% in the 0.2-0.5 micro m range with a theoretical model that takes into account the finite conductivity and roughness of the two metals. The observed discrepancies, which are much larger than the experimental precision, can be attributed to a lack of a complete characterization of the optical properties of the specific samples used in the experiment. PMID- 12906585 TI - Exact nonstationary probabilities in the asymmetric exclusion process on a ring. AB - The complete solution of the master equation for a system of interacting particles of finite density is presented. By using a new form of the Bethe ansatz, the totally asymmetric exclusion process on a ring is solved for arbitrary initial conditions and time intervals. PMID- 12906586 TI - Stress-energy tensor for a massless spin 1/2 field in static black hole spacetimes. AB - The stress-energy tensor for the massless spin 1/2 field is numerically computed outside and on the event horizons of both charged and uncharged static nonrotating black holes, corresponding to the Schwarzschild, Reissner-Nordstrom, and extreme Reissner-Nordstrom solutions of Einstein's equations. The field is assumed to be in a thermal state at the black hole temperature. Comparison is made between the numerical results and previous analytic approximations for the stress-energy tensor in these spacetimes. For the Schwarzschild (charge zero) solution, it is shown that the stress energy differs even in sign from the analytic approximation. For the Reissner-Nordstrom and extreme Reissner-Nordstrom solutions, divergences predicted by the analytic approximations are shown not to exist. PMID- 12906588 TI - Lattice calculation of the lowest-order hadronic contribution to the muon anomalous magnetic moment. AB - We present a quenched lattice calculation of the lowest order [O(alpha(2))] hadronic contribution to the anomalous magnetic moment of the muon which arises from the hadronic vacuum polarization. A general method is presented for computing entirely in Euclidean space, obviating the need for the usual dispersive treatment which relies on experimental data for e(+)e(-) annihilation to hadrons. While the result is not yet of comparable precision to those state-of the-art calculations, systematic improvement of the quenched lattice computation to this level is straightforward and well within the reach of present computers. Including the effects of dynamical quarks is conceptually trivial; the computer resources required are not. PMID- 12906589 TI - Polarization transfer in the 4He(e-->,e'p-->)3H reaction up to Q2=2.6 (GeV/c)2. AB - We have measured the proton recoil polarization in the 4He(e-->,e(')p-->)4H reaction at Q(2)=0.5, 1.0, 1.6, and 2.6 (GeV/c)(2). The measured ratio of polarization transfer coefficients differs from a fully relativistic calculation, favoring the inclusion of a medium modification of the proton form factors predicted by a quark-meson coupling model. In addition, the measured induced polarizations agree reasonably well with the fully relativistic calculation indicating that the treatment of final-state interactions is under control. PMID- 12906590 TI - Net baryon density in Au+Au collisions at the relativistic heavy ion collider. AB - We calculate the net-baryon rapidity distribution in Au+Au collisions at the Relativistic Heavy Ion Collider (RHIC) in the framework of the parton cascade model (PCM). Parton rescattering and fragmentation leads to a substantial increase in the net-baryon density at midrapidity over the density produced by initial primary parton-parton scatterings. The PCM is able to describe the measured net-baryon density at RHIC. PMID- 12906592 TI - Recoil-free spectroscopy of neutral Sr atoms in the Lamb-Dicke regime. AB - Recoil-free as well as Doppler-free spectroscopy was demonstrated on the 1S0-3P1 transition of Sr atoms confined in a one-dimensional optical lattice. By investigating the wavelength and polarization dependence of the ac Stark shift acting on the 1S0 and 3P1(m(J)=0) states, we determined the wavelength where the Stark shifts for both states coincide. This Stark-free optical lattice, allowing the purturbation-free spectroscopy of trapped atoms, may keep neutral-atom based optical standards competitive with single-ion standards. PMID- 12906591 TI - Significance of the fragmentation region in ultrarelativistic heavy-ion collisions. AB - We present measurements of the pseudorapidity distribution of primary charged particles produced in Au+Au collisions at three energies, sqrt[s(NN)]=19.6, 130, and 200 GeV, for a range of collision centrali-ties. The distribution narrows for more central collisions and excess particles are produced at high pseudorapidity in peripheral collisions. For a given centrality, however, the distributions are found to scale with energy according to the "limiting fragmentation" hypothesis. The universal fragmentation region described by this scaling grows in pseudorapidity with increasing collision energy, extending well away from the beam rapidity and covering more than half of the pseudorapidity range over which particles are produced. This approach to a universal limiting curve appears to be a dominant feature of the pseudorapidity distribution and therefore of the total particle production in these collisions. PMID- 12906593 TI - Dramatic nondipole effects in low-energy photoionization: experimental and theoretical study of Xe 5s. AB - The Xe 5s nondipole photoelectron parameter gamma is obtained experimentally and theoretically from threshold to approximately 200 eV photon energy. Significant nondipole effects are seen even in the threshold region of this valence shell photoionization. In addition, contrary to previous understanding, clear evidence of interchannel coupling among quadrupole photoionization channels is found. PMID- 12906594 TI - Model-independent quantum approach for intense laser detachment of a weakly bound electron. AB - We present a model-independent theory for laser detachment of a weakly bound electron having a nonzero angular momentum. Our treatment reduces to the well known Keldysh result for tunnel ionization upon neglecting rescattering effects. Numerical results for the above-threshold detachment spectrum of a negative ion having an outer p electron show significant modification of the rescattering plateau as compared to that for an ion having an outer s electron. PMID- 12906595 TI - Charge driven fragmentation of nucleobases. AB - We studied multiple ionization of single nucleobases by means of slow highly charged ions (Xe(q+), q=5-25). The products of the subsequent fragmentation were studied using high resolution coincidence time-of-flight spectrometry. We observed a strong dependence of the fragment kinetic energies on the initial charge state of the intermediate parent ions as well as on the initial chemical environment of the respective fragment ions within the parent molecule. The data allow us to shed light on the charge distribution within the molecule as well as on the fragmentation dynamics of these intermediate size systems. PMID- 12906596 TI - Theory of an entanglement laser. AB - We consider the creation of polarization entangled light from parametric down conversion driven by an intense pulsed pump field inside a cavity. The multiphoton states produced are close approximations to singlet states of two very large spins. A criterion is derived to quantify the entanglement of such states. We study the dynamics of the system in the presence of losses and other imperfections, concluding that the creation of strongly entangled states with photon numbers up to a million seems achievable. PMID- 12906597 TI - Midband dissipative spatial solitons. AB - We show dissipative spatial solitons in nonlinear optical microresonators in which the refractive index is laterally modulated. In addition to "normal" and "staggered" dissipative solitons, similar to those in spatially modulated conservative systems, a narrow "midband" soliton is shown, having no counterparts in conservative systems. PMID- 12906587 TI - Measurements of the branching fractions and bounds on the charge asymmetries of charmless three-body charged B decays. AB - We present measurements of branching fractions and charge asymmetries for charmless B-meson decays to three-body final states of charged pions and kaons. The analysis uses 81.8 fb(-1) of data collected at the Upsilon(4S) resonance with the BABAR detector at the SLAC PEP-II asymmetric B Factory. We measure the branching fractions B(B+-->pi(+)pi(-)pi(+))=(10.9+/-3.3+/-1.6)x10(-6), B(B+- >K+pi(-)pi(+))=(59.1+/-3.8+/-3.2)x10(-6), and B(B+-->K+K-K+)=(29.6+/-2.1+/ 1.6)x10(-6) and provide 90% C.L. upper limits for other decays. We observe no charge asymmetries for these modes. PMID- 12906598 TI - Optical manipulation of microscale fluid flow. AB - A novel optical method is used both to probe and to control dynamics in experiments on the spreading of microscale liquid films over solid substrates. The flow is manipulated by thermally induced surface-tension gradients that are regulated by controlling the absorption of light in the substrate. This approach permits, for the first time, the measurement of the dispersion relation for the well-known contact line instability; the measurements are compared with theoretical predictions from the slip model for spreading films. The experiments also demonstrate the use of feedback control to suppress instability. These results show that optical control can provide dynamically reconfigurable manipulations of fluid flow, thereby suggesting a general approach for constructing reprogrammable microfluidic devices. PMID- 12906599 TI - Suspension and fall of heavy particles in random two-dimensional flow. AB - We investigate the settling of heavy particles in a steady, two-dimensional random velocity field, and find instances in which particle suspension occurs. This leads to a bimodal velocity distribution that may explain some apparently conflicting results reported in the literature. The bimodal distribution is typically smeared out by a time dependence of the ambient flow but, if the time variation is slow, the settling rates of some particles will be as well. The resulting broadbanded velocity distribution of the settling particles will have significance for processes such as rain drop formation, in which the spread of particle velocities affects the statistics of particle collisions. PMID- 12906600 TI - Positron plasma diagnostics and temperature control for antihydrogen production. AB - Production of antihydrogen atoms by mixing antiprotons with a cold, confined, positron plasma depends critically on parameters such as the plasma density and temperature. We discuss nondestructive measurements, based on a novel, real-time analysis of excited, low-order plasma modes, that provide comprehensive characterization of the positron plasma in the ATHENA antihydrogen apparatus. The plasma length, radius, density, and total particle number are obtained. Measurement and control of plasma temperature variations, and the application to antihydrogen production experiments are discussed. PMID- 12906601 TI - Kolmogorov-Kraichnan scaling in the inverse energy cascade of two-dimensional plasma turbulence. AB - Turbulence in plasmas that are magnetically confined, such as tokamaks or linear devices, is two dimensional or at least quasi two dimensional due to the strong magnetic field, which leads to extreme elongation of the fluctuations, if any, in the direction parallel to the magnetic field. These plasmas are also compressible fluid flows obeying the compressible Navier-Stokes equations. This Letter presents the first comprehensive scaling of the structure functions of the density and velocity fields up to 10th order in the PISCES linear plasma device and up to 6th order in the Mega-Ampere Spherical Tokamak (MAST). In the two devices, it is found that the scaling of the turbulent fields is in good agreement with the prediction of the Kolmogorov-Kraichnan theory for two dimensional turbulence in the energy cascade subrange. PMID- 12906602 TI - Spin precession waves in superfluid 3He-B. AB - Spin precession waves of homogeneously precessing domains (HPD) in superfluid 3He B have been studied at 11 bars and temperatures down to 0.45T(c). The waves were excited by an alternating longitudinal magnetic field with an axial symmetry, applied as a small perturbation ranging from 1 nT up to a few micro T. When the spin precession wave is excited, two nuclear magnetic resonances simultaneously coexist: first, the high frequency resonance used for excitation of the HPD, and, second, the low-frequency resonance of the HPD wave mode. We report the first experimental evidence of the nonlinear behavior of low-frequency precession spin wave modes of the continuously maintained HPD. PMID- 12906603 TI - Elastic deformation of polycrystals. AB - We propose a framework to model elastic properties of polycrystals by coupling crystal orientational degrees of freedom with elastic strains. Our model encodes crystal symmetries and takes into account explicitly the strain compatibility induced long-range interaction between grains. The coupling of crystal orientation and elastic interactions allows for the rotation of individual grains by an external load. We apply the model to simulate uniaxial tensile loading of a 2D polycrystal within linear elasticity and a system with elastic anharmonicities that describe structural phase transformations. We investigate the constitutive response of the polycrystal and compare it to that of single crystals with crystallographic orientations that form the polycrystal. PMID- 12906604 TI - Griffiths singularities in unbinding of strongly disordered polymers. AB - Griffiths singularities occurring in the unbinding of strongly disordered heteropolymers are studied. A model with two randomly distributed binding energies, -1 and -v, is introduced and studied analytically by analyzing the Lee Yang zeros of the partition sum. It is demonstrated that in the limit v--> infinity the model exhibits a Griffiths type singularity at a temperature T(G)=O(1) corresponding to melting of long homogeneous domains of the low binding energy. For finite v>>1 the model is expected to exhibit an additional, unbinding, transition at a high temperature T(M)=O(v). PMID- 12906605 TI - Thermal motion and energetics of self-assembled domain structures: Pb on Cu(111). AB - Low energy electron microscope measurements of the thermal motion of 50-200 nm diameter Pb islands on Cu(111) are used to establish the nature and determine the strength of interactions that give rise to self-assembly in this two-dimensional, two-phase system. The results show that self-assembled patterns arise from a temperature-independent surface stress difference of approximately 1.2 N/m between the two phases. With increasing Pb coverage, the domain patterns evolve in a manner consistent with models based on dipolar repulsions caused by elastic interactions due to a surface stress difference. PMID- 12906606 TI - Finite-size scaling analysis of the glass transition. AB - We show that finite-size scaling techniques can be employed to study the glass transition. Our results follow from the postulate of a diverging dynamical correlation length at the glass transition whose physical manifestation is the presence of dynamical heterogeneities. We introduce a parameter B(T,L) whose temperature, T, and system size, L, dependences permit a precise location of the glass transition. We discuss the finite-size scaling behavior of a diverging susceptibility chi(L,T). These new techniques are successfully used to study two lattice models. The analysis straightforwardly applies to any glass-forming system. PMID- 12906607 TI - Field-induced formation of nanometer-sized water bridges. AB - A method to form and manipulate the properties of nanometer-size liquid bridges by an external electric field is discussed. The properties of bridges are shown to be the result of an interplay among the field-induced polarization of the water layer adsorbed on the surface, the surface energy, and the water condensation from the humid air. For a given tip-sample separation, a simple model predicts the existence of a threshold voltage V(th) to form the bridge in full agreement with experiments. PMID- 12906608 TI - Identification of step atoms by high resolution core level spectroscopy. AB - Vicinal Rh(111) surfaces are studied with high resolution core level photoemission. We demonstrate the possibility to distinguish between the different kinds of surface atoms on these surfaces by virtue of their 3d core level binding energies. In particular, the low coordinated step atoms are found to exhibit a clear fingerprint in Rh 3d spectra. We demonstrate how this may be used to show that initial oxygen adsorption occurs on the steps and not on the terraces of the vicinal surfaces. PMID- 12906609 TI - Stacking-fault nucleation on Ir(111). AB - Variable temperature scanning tunneling microscopy experiments reveal that in Ir(111) homoepitaxy islands nucleate and grow both in the regular fcc stacking and in the faulted hcp stacking. Analysis of this effect in dependence on deposition temperature leads to an atomistic model of stacking-fault formation: The large, metastable stacking-fault islands grow by sufficiently fast addition of adatoms to small mobile adatom clusters which occupy in thermal equilibrium the hcp sites with a significant probability. Using parameters derived independently by field ion microscopy, the model accurately describes the results for Ir(111) and is expected to be valid also for other surfaces. PMID- 12906610 TI - Curvature-dependent surface tension of a growing droplet. AB - A ghost interface simulation technique is developed and applied to supersaturated Lennard-Jones liquid-vapor interfaces. It is shown that the surface tension decreases approximately linearly with the supersaturation ratio and that it vanishes at the spinodal. The effect leads to a curvature-dependent surface tension since, it is argued, the local supersaturation of the vapor above a droplet is greater than in the bulk due to slow diffusion in the vapor phase. An analytic approximation is given for the local supersaturation ratio, and an analytic expression for this contribution to Tolman's length is derived. The theory gives a smaller critical radius and reduces the free energy barrier to nucleation compared to classical homogeneous nucleation theory, which have important implications for the kinetics of droplet and bubble formation. PMID- 12906611 TI - Coexistence of s-wave superconductivity and antiferromagnetism. AB - We study the phase diagram of a new model that exhibits a first order transition between s-wave superconducting and antiferromagnetic phases. The model, a generalized Hubbard model augmented with competing spin-spin and pair-pair interactions, was investigated using the projector quantum Monte Carlo method. Upon varying the Hubbard U from attractive to repulsive, we find a first order phase transition between superconducting and antiferromagnetic states. PMID- 12906612 TI - Parameter-free calculation of response functions in time-dependent density functional theory. AB - We have established and implemented a fully ab initio method which allows one to calculate optical absorption spectra, including excitonic effects, without solving the cumbersome Bethe-Salpeter equation, but obtaining results of the same precision. This breakthrough has been achieved in the framework of time-dependent density-functional theory, using new exchange-correlation kernels f(xc) that are free of any empirical parameter. We show that the same excitonic effects in the optical spectra can be reproduced through different f(xc)'s, ranging from frequency-dependent ones to a static one, by varying the kernel's spatial degrees of freedom. This indicates that the key quantity is not f(xc), but f(xc) combined with a response function. We present results for the optical absorption of bulk Si and SiC in good agreement with experiment, almost indistinguishable from those of the Bethe-Salpeter approach. PMID- 12906613 TI - Change of electronic structure in Ca2RuO4 induced by orbital ordering. AB - Optical conductivity spectra sigma(omega) were used to investigate the effect of orbital ordering on the electronic structure of Ca2RuO4. Our LDA+U calculation predicts Ru 4d(xy) ferro-orbital ordering at the ground state, and well explains the present sigma(omega) as well as the reported O 1s x-ray absorption spectra. Variation of temperature (T) causes a large change of spectral weight over several eV as well as collapse of a charge gap accompanied by elongation of the c axis Ru-O bond length. These results clearly indicate that the d(xy) orbital ordering plays a crucial role in the metal-insulator transition and the T dependent electronic structure on a large energy scale. PMID- 12906614 TI - Pseudopotential theory of Auger processes in CdSe quantum dots. AB - Auger rates are calculated for CdSe colloidal quantum dots using atomistic empirical pseudopotential wave functions. We predict the dependence of Auger electron cooling on size, on correlation effects (included via configuration interaction), and on the presence of a spectator exciton. Auger multiexciton recombination rates are predicted for biexcitons as well as for triexcitons. The results agree quantitatively with recent measurements and offer new predictions. PMID- 12906615 TI - Nonperturbative solution for BLOCH electrons in constant magnetic fields. AB - A general theoretical approach for the nonperturbative Bloch solution of Schrodinger's equation in the presence of a constant magnetic field is presented. Using a singular gauge transformation based on a lattice of magnetic flux lines, an equivalent quantum system with a periodic vector potential is obtained. For rational magnetic fields this system forms a magnetic superlattice for which Bloch's theorem then applies. Extensions of the approach to particles with spin and many-body systems and connections to the theory of magnetic translation groups are discussed. PMID- 12906616 TI - Giant Nernst effect and lock-in currents at magic angles in (TMTSF)2PF6. AB - We have measured the thermoelectric signal along the a axis in (TMTSF)2PF6 at 10 kbar as a function of the orientation of the applied magnetic field. Resonantlike Nernst signals were found with a dramatic sign change as the field was rotated through the "Lebed magic angles." The sign change indicates that the electrical current is "locked in" to the magic angle (interchain) directions for field alignment close to, but on either side of, the magic angles. The amplitude of signals near these angles is many orders of magnitude larger than expected from conventional Boltzmann transport theory. PMID- 12906617 TI - Direct measurement of the spin polarization of the magnetic semiconductor (Ga,Mn)As. AB - We have carried out a direct measurement of the degree of spin polarization (P) of the magnetic semiconductor Ga1-xMnxAs using Andreev reflection spectroscopy. Analyses of the conductance spectra of high transparency Ga(0.95)Mn(0.05)As/Ga junctions consistently yield an intrinsic value for P greater than 85%. Our experiments also revealed an extreme sensitivity of the measured spin polarization to the nature and quality of the interface for this material. PMID- 12906618 TI - Variable-range hopping and quantum creep in one dimension. AB - We study the quantum nonlinear response to an applied electric field E of a one dimensional pinned charge-density wave or Luttinger liquid in the presence of disorder. From an explicit construction of low-lying metastable states and of bounce instanton solutions between them, we demonstrate quantum creep v=e( c/E(1/2)) as well as a sharp crossover at E=E(*) towards a linear response form consistent with variable-range hopping arguments, but dependent only on electronic degrees of freedom. PMID- 12906619 TI - Charge transport in the insulating state of (DMe-DCNQI)2Li above T(SP): a possible fractional charge soliton conduction with +/-1 / 2e. AB - A spin-Peierls system (DMe-DCNQI)2Li is studied with W-band electron paramagnetic resonance (EPR) ( approximately 94 GHz) to unveil a charge transport mechanism in the insulating 4k(F) charge density wave state above T(SP). The electron hopping between the neighbor DCNQI columns provides an additional broadening of the EPR linewidth, since the neighbor columns are generally nonequivalent to each other with respect to g shift. The obtained intercolumn hopping rates lead us to the conclusion that the electron hopping to a hole soliton carrying a fractional charge of e / 2 in the neighbor column dominates the intercolumn charge transport. PMID- 12906620 TI - Effects of spin and exchange interaction on the Coulomb-blockade peak statistics in quantum dots. AB - We derive a closed expression for the linear conductance through a quantum dot in the Coulomb-blockade regime in the presence of a constant exchange interaction. With this expression we calculate the temperature dependence of the conductance peak-height and peak-spacing statistics in chaotic quantum dots. Using a realistic value of the exchange interaction, we find significantly better agreement with experimental data as compared with the statistics obtained in the absence of exchange. PMID- 12906621 TI - Quantum interference in carbon-nanotube electron resonators. AB - A new mechanism is proposed to explain the slow conductance fluctuations in the conductance-gate voltage plot observed in the nanotube electron resonators. It is found that the slow conductance fluctuation is an intrinsic quantum interference phenomenon and exists in all metallic nanotube resonators except zigzag ones. Analytical expressions for both slow and rapid oscillation periods of the conductance fluctuations have been derived, which are well consistent with the existing experiments. It is predicted that the ratio of the slow oscillation period to the rapid one is independent of the gate-voltage efficiency, and determined only by the nanotube length used in experiments. PMID- 12906622 TI - Dynamical symmetry breaking as the origin of the zero-dc-resistance state in an ac-driven system. AB - Under a strong ac drive the zero-frequency linear response dissipative resistivity rho(d)(j=0) of a homogeneous state is allowed to become negative. We show that such a state is absolutely unstable. The only time-independent state of a system with a rho(d)(j=0)<0 is characterized by a current which almost everywhere has a magnitude j(0) fixed by the condition that the nonlinear dissipative resistivity rho(d)(j(2)(0))=0. As a result, the dissipative component of the dc-electric field vanishes. The total current may be varied by rearranging the current pattern appropriately with the dissipative component of the dc electric field remaining zero. This result, together with the calculation of Durst et al., indicating the existence of regimes of applied ac microwave field and dc magnetic field where rho(d)(j=0)<0, explains the zero-resistance state observed by Mani et al. and Zudov et al. PMID- 12906623 TI - Low-energy spectra in t-J-type models at low doping levels. AB - Based on a variational approach, we propose that there are two kinds of low energy states in the t-J-type models at low doping. In a quasiparticle state an unpaired spin bound to a hole with a well-defined momentum can be excited with spin waves. The resulting state shows a suppression of antiferromagnetic order around the hole with the profile of a spin bag. These spin-bag states with spin and charge or hole separated form a continuum of low-energy excitations. Very different properties predicted by these two kinds of states explain a number of anomalous results observed in the exact diagonalization studies on small clusters up to 32 sites. PMID- 12906624 TI - Strain and high temperature superconductivity: unexpected results from direct electronic structure measurements in thin films. AB - Angle-resolved photoemission spectroscopy reveals very surprising strain-induced effects on the electronic band dispersion of epitaxial La(2-x)Sr(x)CuO(4-delta) thin films. In strained films we measure a band that crosses the Fermi level (E(F)) well before the Brillouin zone boundary. This is in contrast to the flat band reported in unstrained single crystals and in our unstrained films, as well as in contrast to the band flattening predicted by band structure calculations for in-plane compressive strain. In spite of the density of states reduction near E(F), the critical temperature increases in strained films with respect to unstrained samples. These results require a radical departure from commonly accepted notions about strain effects on high temperature superconductors, with possible general repercussions on superconductivity theory. PMID- 12906625 TI - Variable electrostatic transformer: controllable coupling of two charge qubits. AB - We propose and investigate a novel method for the controlled coupling of two Josephson charge qubits by means of a variable electrostatic transformer. The value of the coupling capacitance is given by the discretized curvature of the lowest energy band of a Josephson junction, which can be positive, negative, or zero. We calculate the charging diagram of the two-qubit system that reflects the transition from positive to negative through vanishing coupling. We also discuss how to implement a phase gate making use of the controllable coupling. PMID- 12906626 TI - Elasticity-driven nanoscale electronic structure in superconductors. AB - The effects of long-range anisotropic elastic deformations on electronic structure in superconductors are analyzed within the framework of the Bogoliubov de Gennes equations. Cases of twin boundaries and isolated defects are considered as illustrations. We find that the superconducting order parameter is depressed in the regions where pronounced lattice-deformation occurs. The calculated local density of states suggests that the electronic structure is strongly modulated in response to lattice deformations, and propagates to longer distances. In particular, this allows the trapping of low-lying quasiparticle states around defects. Some of our predictions can be directly tested by scanning tunneling microscopy experiments. PMID- 12906627 TI - Multiple Andreev reflections in a carbon nanotube quantum dot. AB - We report resonant multiple Andreev reflections in a multiwall carbon nanotube quantum dot coupled to superconducting leads. The position and magnitude of the subharmonic gap structure is found to depend strongly on the level positions of the single-electron states which are adjusted with a gate electrode. We discuss a theoretical model of the device and compare the calculated differential conductance with the experimental data. PMID- 12906628 TI - Sound attenuation study on the bose-Einstein condensation of magnons in TlCuCl3. AB - We investigate experimentally and theoretically sound attenuation in the quantum spin system TlCuCl3 in magnetic fields at low temperatures. Near the point of Bose-Einstein condensation of magnons a sharp peak in the sound attenuation is observed. The peak demonstrates a hysteresis as a function of the magnetic field pointing to a first-order contribution to the transition. The sound damping has a Drude-like form arising as a result of hard-core magnon-magnon collisions. The strength of the coupling between lattice and magnons is estimated from the experimental data. The puzzling relationship between the transition temperature and the concentration of magnons is explained by their "relativistic" dispersion. PMID- 12906629 TI - Anomalous Hall effect in ferromagnetic semiconductors in the hopping transport regime. AB - We present a theory of the anomalous Hall effect in ferromagnetic (Ga,Mn)As in the regime when conduction is due to phonon-assisted hopping of holes between localized states in the impurity band. We show that the microscopic origin of the anomalous Hall conductivity in this system can be attributed to a phase that a hole gains when hopping around closed-loop paths in the presence of spin-orbit interactions and background magnetization of the localized Mn moments. Mapping the problem to a random resistor network, we derive an analytic expression for the macroscopic anomalous Hall conductivity sigma(AH)(xy). We show that sigma(AH)(xy) is proportional to the first derivative of the density of states varrho(epsilon) and thus can be expected to change sign as a function of impurity band filling. We also show that sigma(AH)(xy) depends on temperature as the longitudinal conductivity sigma(xx) within logarithmic accuracy. PMID- 12906630 TI - Ultrafast optical switching to a metallic state by photoinduced mott transition in a halogen-bridged nickel-chain compound. AB - We demonstrate the ultrafast photoinduced Mott transition from a charge transfer insulator to a metal in a halogen-bridged Ni-chain compound by pump-probe reflection spectroscopy. Upon the irradiation of a 130-femtosecond laser pulse, the spectral weight of the gap transition is transferred to the inner-gap region. When the photoexcitation density exceeds 0.1/Ni site, the Drude-like high reflection band appears in the infrared region, signaling the formation of a metallic state. The photogeneration of the metallic state and the subsequent recovery to the original gapped state occur within a few picoseconds. PMID- 12906631 TI - Polaritonic stop-band transparency via exciton-biexciton coupling in CuCl. AB - Radiation is almost completely reflected within the exciton-polariton stop band of a semiconductor, as in the typical case of CuCl. We predict, however, that a coherently driven exciton-biexciton transition allows for the propagation of a probe light beam within the stop band. The phenomenon is reminiscent of electromagnetically induced transparency effects occurring in three-level atomic systems, except that it here involves delocalized electronic excitations in a crystalline structure via a frequency and wave-vector selective polaritonic mechanism. A well-developed transparency, favored by the narrow linewidth of the biexciton, is established within the stop band where a probe pulse may propagate with significant delays. The transparency window can be controlled via the pump beam detuning and intensity. PMID- 12906632 TI - Comparison of electron and neutron compton scattering from entangled protons in a solid polymer. AB - We present, for the first time, a direct comparison between electron (ECS) and neutron (NCS) Compton scattering results from protons of a solid polymer. The momentum distributions of hydrogen obtained from ECS and NCS are in excellent agreement. In both experiments, a strong "anomalous" shortfall in the scattering intensity of protons [first detected in liquid water with NCS [Phys. Rev. Lett. 79, 2839 (1997)]]] is found ranging from about 20% up to 50%, depending on the momentum transfer applied. The characteristic times of electron- and neutron proton collisions lie in the subfemtosecond range. The presented ECS and NCS results provide further direct evidence for this striking effect, which has been ascribed to attosecond quantum entanglement of the protons. PMID- 12906633 TI - First-principles determination of electromechanical responses of solids under finite electric fields. AB - We describe a first-principles, easy-to-implement, and efficient approach for determining the structural geometry of insulating solids under finite electric fields. This method consists of simultaneously minimizing the field-induced total ionic forces and the electric free energy. Moreover, we present a theory to analyze its predictions that provides a microscopic understanding of electro mechanical responses in materials. We illustrate this approach by computing piezoelectric and dielectric responses of two rather different compounds, namely, ferroelectric PbTiO3 and semiconductor GaN. PMID- 12906634 TI - Quantum key distribution with high loss: toward global secure communication. AB - We propose a decoy-pulse method to overcome the photon-number-splitting attack for Bennett-Brassard 1984 quantum key distribution protocol in the presence of high loss: A legitimate user intentionally and randomly replaces signal pulses by multiphoton pulses (decoy pulses). Then they check the loss of the decoy pulses. If the loss of the decoy pulses is abnormally less than that of signal pulses, the whole protocol is aborted. Otherwise, to continue the protocol, they estimate the loss of signal multiphoton pulses based on that of decoy pulses. This estimation can be done with an assumption that the two losses have similar values. We justify that assumption. PMID- 12906635 TI - Majorization criterion for distillability of a bipartite quantum state. AB - Bipartite quantum states are classified into three categories: separable states, bound entangled states, and free entangled states. It is of great importance to characterize these families of states for the development of quantum information science. In this Letter, I show that the separable states and the bound entangled states have a common spectral property. More precisely, I prove that for undis tillable-separable and bound entangled-states, the eigenvalue vector of the global system is majorized by that of the local system. This result constitutes a new sufficient condition for distillability of bipartite quantum states. This is achieved by proving that if a bipartite quantum state satisfies the reduction criterion for distillability, then it satisfies the majorization criterion for separability. PMID- 12906636 TI - Lateral scaling in carbon-nanotube field-effect transistors. AB - We have fabricated carbon-nanotube (CN) field-effect transistors with multiple, individually addressable gate segments. The devices exhibit markedly different transistor characteristics when switched using gate segments controlling the device interior versus those near the source and drain. We ascribe this difference to a change from Schottky-barrier modulation at the contacts to bulk switching. We also find that the current through the bulk portion is independent of gate length for any gate voltage, offering direct evidence for ballistic transport in semiconducting carbon nanotubes over at least a few hundred nanometers, even for relatively small carrier velocities. PMID- 12906637 TI - Superlattice Turing structures in a photosensitive reaction-diffusion system. AB - Families of complex superlattice structures, consisting of combinations of basic hexagonal or square patterns, are found in a photosensitive reaction-diffusion system. The structures are induced by simple illumination patterns whose wavelengths are appropriately related to that of the system's intrinsic Turing pattern. Computer simulations agree with the structures and their stability. The technique offers a general approach to generating superlattices for use in information storage and other applications. PMID- 12906638 TI - Defect-mediated turbulence in systems with local deterministic chaos. AB - Defect-mediated turbulence is shown to exist in media where the underlying local dynamics is deterministically chaotic. While many of the characteristics of defect-mediated turbulence, such as the exponential decay of correlations and a squared Poissonian distribution for the number of defects, are identical to those seen in oscillatory media, the fluctuations in the number of defects differ significantly. The power spectra suggest the existence of underlying correlations that lead to a different and nonuniversal scaling structure in chaotic media. PMID- 12906639 TI - Geometric theory of diblock copolymer phases. AB - We analyze the energetics of spherelike micellar phases in diblock copolymers in terms of well-studied, geometric quantities for their lattices. We argue that the A15 lattice with Pm3;n symmetry should be favored as the blocks become more symmetric and corroborate this through a self-consistent field theory. Because phases with columnar or bicontinuous topologies intervene, the A15 phase, though metastable, is not an equilibrium phase of symmetric diblocks. We investigate the phase diagram of branched diblocks and find that the A15 phase is stable. PMID- 12906640 TI - Replica-exchange method using the generalized effective potential. AB - We propose an effective scheme for fast conformational searches by combining the replica exchange method (REM) with the generalized effect potential concept. The present method introduces the "q" value from the effective potential as a coupling parameter. It is found that the new method not only requires a much smaller number of replicas than the conventional REM, but also makes it possible to perform effective conformational sampling of complex systems with correct distributions maintained. The advantage of the present method has been demonstrated with in vacuo alanine dipeptide using a molecular dynamics simulation. PMID- 12906641 TI - Is earthquake triggering driven by small earthquakes? AB - Using a catalog of seismicity for Southern California, we measure how the number of triggered earthquakes increases with the earthquake magnitude. The trade-off between this relation and the distribution of earthquake magnitudes controls the relative role of small compared to large earthquakes. We show that seismicity triggering is driven by the smallest earthquakes, which trigger fewer events than larger earthquakes, but which are much more numerous. We propose that the nontrivial scaling of the number of triggered earthquakes emerges from the fractal spatial distribution of seismicity. PMID- 12906642 TI - Impact of cosmic rays on stratospheric chlorine chemistry and ozone depletion. AB - Dissociation induced by cosmic rays of chlorofluorocarbons (CFC) and HCl on the surfaces of polar stratospheric clouds (PSC) has been suggested as playing a significant role in causing the ozone hole. However, observed stratospheric CFC distributions are inconsistent with a destruction of CFC on PSC surfaces and no significant correlation exists between ozone levels and cosmic-ray activity inside the polar regions. Model simulations indicate that this mechanism can have only a limited impact on chemical ozone loss and thus on the recovery of stratospheric ozone. PMID- 12906644 TI - Comment on "vibrational recognition of hydrogen-bonded water networks on a metal surface". PMID- 12906643 TI - Probabilistic prediction in scale-free networks: diameter changes. AB - In complex systems, responses to small perturbations are too diverse to definitely predict how much they would be, and then such diverse responses can be predicted in a probabilistic way. Here we study such a problem in scale-free networks, for example, the diameter changes by the deletion of a single vertex for various in silico and real-world scale-free networks. We find that the diameter changes are indeed diverse and their distribution exhibits an algebraic decay with an exponent zeta asymptotically. Interestingly, the exponent zeta is robust as zeta approximately 2.2(1) for most scale-free networks and insensitive to the degree exponents gamma as long as 2e(+)e(-) annihilation with a modified rho propagator. They may be linked to chiral symmetry restoration and support the notion that the in-medium modifications of the rho are more driven by baryon density than by temperature. PMID- 12906653 TI - Observation of trielectronic recombination in Be-like Cl ions. AB - Recombination involving the core excitation of two electrons, which may be termed trielectronic recombination, has been experimentally identified for the first time. Using Cl13+ ions circulating in the TSR heavy-ion storage ring, we have observed surprisingly strong low-energy trielectronic recombination resonances, comparable to the dielectronic process. At higher electron-ion collision energies, trielectronic recombination is suppressed due to the autoionization of the triply excited intermediate state into excited final states. The formation of the intermediate state depends sensitively on configuration mixing, making trielectronic recombination a challenge to atomic-structure calculations. PMID- 12906654 TI - Photoemission and ionization of He+ under simultaneous irradiation of fundamental laser and high-order harmonic pulses. AB - We theoretically study the response of He+ exposed simultaneously to an intense Ti:sapphire laser and its 27th or 13th harmonic pulses. High-order harmonic emission from He+ is enhanced by many orders of magnitude compared with the case of the fundamental pulse alone. Moreover, while an individual 10 fs laser (wavelength lambda(F)=800 nm) or its 27th harmonic pulse with a peak intensity of 3 x 10(14) and 10(13) W/cm(2), respectively, ionizes no more than 5 x 10(-6) of He+, their combined pulse leads to a surprisingly high He2+ yield of 17%. PMID- 12906655 TI - Nonadiabatic alignment of asymmetric top molecules: field-free alignment of iodobenzene. AB - Nonadiabatic laser alignment of an asymmetric top molecule is studied using the combination of a quantum dynamical theory and time-resolved photofragment imaging experiments. In particular, the degree of alignment of iodobenzene, induced by an intense, linearly polarized picosecond laser pulse, is calculated and measured. Pronounced alignment is obtained under field-free conditions. PMID- 12906656 TI - Hyperfine splitting, isotope shift, and level energy of the 3S states of (6,7)Li. AB - We study the 2S-3S transition of (6,7)Li by high-precision laser spectroscopy using two-photon Doppler-free excitation and photoionization detection. Interferometric cross referencing to metrologic Rb 3S-5D two-photon transitions allowed measurement of the transition isotope shift and hyperfine splitting in the 3S state with precision at the 30 kHz level. The results are IS=11 453.734(30) MHz, A(3S)(6Li)=35.263(15) MHz, and A(3S)(7Li)=93.106(11) MHz. Combined with recent theoretical work, the isotope shift yields a new value for the change in squared nuclear charge radii DeltaR(2)=0.47(5) fm(2). This is compared with other work and some existing discrepancies are resolved. PMID- 12906657 TI - Photoelectron imaging of helium droplets. AB - The photoionization and photoelectron spectroscopy of He nanodroplets (10(4) atoms) has been studied by photoelectron imaging with photon energies from 22.5 24.5 eV. Total electron yield measurements reveal broad features, whose onset is approximately 1.5 eV below the ionization potential of atomic He. The photoelectron spectra are dominated by very low energy electrons, with less than 0.6 meV. These results are attributed to the formation and autoionization of highly vibrationally excited He(*)(n) Rydberg states within the cluster, followed by strong final state interactions between the photoelectron and the droplet. PMID- 12906658 TI - Multiple higher-order stop gaps in infrared polymer photonic crystals. AB - Engineering of stop gaps between higher photonic bands provides an alternative to miniaturization of photonic crystals. Femtosecond laser microfabrication of highly correlated void channel polymer microstructures results in photonic crystals with large stop gaps and a multitude of higher-order gaps in the mid- and near-infrared spectral regions. The gap wavelengths obey Bragg's law. Consistent with theory, varying the woodpile structure unit cell allows for tuning the number of higher-order gaps, and transitions from mere resonant Bragg scattering to stop band total reflection are observed. PMID- 12906659 TI - Ideality in a fiber-taper-coupled microresonator system for application to cavity quantum electrodynamics. AB - The ability to achieve near lossless coupling between a waveguide and a resonator is fundamental to many quantum-optical studies as well as to practical applications of such structures. The nature of loss at the junction is described by a figure of merit called ideality. It is shown here that under appropriate conditions ideality in excess of 99.97% is possible using fiber-taper coupling to high-Q silica microspheres. To verify this level of coupling, a technique is introduced that can both measure ideality over a range of coupling strengths and provide a practical diagnostic of parasitic coupling within the fiber-taper waveguide junction. PMID- 12906660 TI - Statistics of multiply scattered broadband terahertz pulses. AB - We describe the first measurements of the diffusion of broadband single-cycle optical pulses through a highly scattering medium. Using terahertz time-domain spectroscopy, we measure the electric field of a multiply scattered wave with a time resolution shorter than one optical cycle. This time-domain measurement provides information on the statistics of both the amplitude and phase distributions of the diffusive wave. We develop a theoretical description, suitable for broadband radiation, which adequately describes the experimental results. PMID- 12906661 TI - Phase imaging and nanoscale currents in phase objects imaged with fast electrons. AB - We derive the magnetic transport-of-intensity equation (MTIE) that links defocused contrast of magnetic nanoobjects imaged by partially coherent electron waves to their micromagnetic parameters. This provides Maxwell's explanation for observable contrast in terms of the Z component of currents existing around vortices in superconductors and domain vortices and walls in ferromagnets. The solution of the MTIE via Fourier transform is used for quantitative mapping of magnetic flux and projected induction in magnetic and superconducting materials imaged by Lorentz microscopy. PMID- 12906662 TI - Physical factors limiting the spectral extent and band gap dependence of supercontinuum generation. AB - Based on simulations of femtosecond pulse propagation in water we elucidate the physical factors that limit the attainable spectral extent of supercontinuum generation in bulk media. In contrast to the standard scenario, where arrest of self-focusing collapse by multiphoton absorption and plasma defocusing are viewed as imposing a limit to supercontinuum broadening, we show that linear chromatic dispersion also plays a major role. This insight provides an intuitive explanation of the observed band gap dependence of the attainable supercontinuum generation in condensed media. PMID- 12906663 TI - Coarse-grained probabilistic automata mimicking chaotic systems. AB - Discretization of phase space usually nullifies chaos in dynamical systems. We show that if randomness is associated with discretization dynamical chaos may survive and be indistinguishable from that of the original chaotic system, when an entropic, coarse-grained analysis is performed. Relevance of this phenomenon to the problem of quantum chaos is discussed. PMID- 12906664 TI - Anomalous heat conduction and anomalous diffusion in one-dimensional systems. AB - We establish a connection between anomalous heat conduction and anomalous diffusion in one-dimensional systems. It is shown that if the mean square of the displacement of the particle is =2Dt(alpha)(01) implies anomalous heat conduction with a divergent thermal conductivity (beta>0). More interestingly, subdiffusion (alpha<1) implies anomalous heat conduction with a convergent thermal conductivity (beta<0), and, consequently, the system is a thermal insulator in the thermodynamic limit. Existing numerical data support our results. PMID- 12906665 TI - Fermat principle for a nonstationary medium. AB - One possible formulation of a variational principle of the Fermat type for systems with time-dependent parameters is suggested. In a stationary case, it reduces to the Mopertui-Lagrange least-action principle. A class of Hamiltonians (dispersion relations) is indicated, for which the variational principle reduces to the Fermat principle in a general nonstationary case. Hamiltonians that are homogeneous functions of momenta are in this category. For the important case of nondispersive waves (corresponding to Hamiltonians being homogeneous function of momenta order 1) the Fermat principle fully determines the geometry of the rays. Equations relating the variation of signal frequency with the rate of change of propagation time are established. PMID- 12906666 TI - Estimating surfactant surface coverage and decomposing its effect on drop deformation. AB - A novel method is introduced to estimate surface coverage and the equation of state of insoluble surfactant on droplets, involving measurement of interfacial tension on a single parent drop and progressively subdivided generations of daughter drops. This has enabled quantitative decomposition of the dilution, tip stretching, and Marangoni effects of surfactants on drop deformation. For a small viscosity ratio of 0.09, the Marangoni effect dominates, increasing first and then decreasing with surface coverage, the dilution effect is significant at high, and tip-stretching only at low surface coverage. For a viscosity ratio of 2.3, the dilution effect dominates, and neither Marangoni nor tip-stretching effects play an important role. PMID- 12906667 TI - Anomalous transport scaling in the DIII-D tokamak matched by supercomputer simulation. AB - Gyrokinetic simulation of tokamak transport has evolved sufficiently to allow direct comparison of numerical results with experimental data. It is to be emphasized that only with the simultaneous inclusion of many distinct and complex effects can this comparison realistically be made. Until now, numerical studies of tokamak microturbulence have been restricted to either (a) flux tubes or (b) electrostatic fluctuations. Using a newly developed global electromagnetic solver, we have been able to recover via direct simulation the Bohm-like scaling observed in DIII-D L-mode discharges. We also match, well within experimental uncertainty, the measured energy diffusivities. PMID- 12906668 TI - Observation of velocity-independent electron transport in the reversed field pinch. AB - Confinement of runaway electrons has been observed for the first time in a reversed field pinch during improved-confinement plasmas in the Madison Symmetric Torus. Energy-resolved hard-x-ray flux measurements have been used to determine the velocity dependence of the electron diffusion coefficient, utilizing computational solutions of the Fokker-Planck transport equation. With improved confinement, the fast electron diffusivity drops by 2 orders of magnitude and is independent of velocity. This suggests a change in the transport mechanism away from stochastic magnetic field diffusion. PMID- 12906669 TI - Novel turbulence trigger for neoclassical tearings mode in tokamaks. AB - A stochastic trigger by microturbulence for a neoclassical tearing mode (NTM) is studied. The NTM induces a topological change of magnetic structure and has a subcritical nature. The transition rate of the probability density function for and statistically averaged amplitude of the NTM are obtained. The boundary in the phase diagram is determined as the statistical long time average of the transition conditions. The NTM can be excited by crossing this boundary even in the absence of other global instabilities. PMID- 12906670 TI - Electron heat transport measured in a stochastic magnetic field. AB - New profile measurements have allowed the electron thermal diffusivity profile to be estimated from power balance in the Madison Symmetric Torus where magnetic islands overlap and field lines are stochastic. The measurements show that (1) the electron energy transport is conductive not convective, (2) the measured thermal diffusivities are in good agreement with numerical simulations of stochastic transport, and (3) transport is greatly reduced near the reversal surface where magnetic diffusion is small. PMID- 12906671 TI - Double reentrance of surface arrangements in freestanding liquid crystal films. AB - The molecular arrangement of tilted surface layers on freestanding films above the bulk smectic-A-smectic-C(*) transition has been studied by null transmission ellipsometry. In the presence of an applied in-plane electric field, a synclinic anticlinic-synclinic-anticlinic double reentrance has been observed. Furthermore, we have characterized a novel transition from the synclinic arrangement to a structure in which the interior layers tilt in the same orientation and the outermost surface layers tilt oppositely with the interior layers. PMID- 12906672 TI - Nucleation of solids in solids: ferrites and martensites. AB - When a solid such as iron is cooled across a structural transition, its final microstructure depends sensitively on the cooling rate and the depth of quench. For instance, an infinitesimally slow cooling or a shallow quench results in an equilibrium "ferrite," while a rapid cooling or a deep quench gives rise to a metastable twinned "martensite." In this paper, we arrive at a single formalism which qualitatively describes the transformation to both a ferrite and a martensite. Fundamental to this understanding is our identification of the crucial dynamical role played by nonelastic degrees of freedom in determining the final microstructure of the product solid. PMID- 12906673 TI - Dynamics of conformal maps for a class of non-Laplacian growth phenomena. AB - Time-dependent conformal maps are used to model a class of growth phenomena limited by coupled non-Laplacian transport processes, such as nonlinear diffusion, advection, and electromigration. Both continuous and stochastic dynamics are described by generalizing conformal-mapping techniques for viscous fingering and diffusion-limited aggregation, respectively. The theory is applied to simulations of advection-diffusion-limited aggregation in a background potential flow. A universal crossover in morphology is observed from diffusion limited to advection-limited fractal patterns with an associated crossover in the growth rate, controlled by a time-dependent effective Peclet number. Remarkably, the fractal dimension is not affected by advection, in spite of dramatic increases in anisotropy and growth rate, due to the persistence of diffusion limitation at small scales. PMID- 12906674 TI - Effects of atomic short-range order on the properties of perovskite alloys in their morphotropic phase boundary. AB - The effects of atomic short-range order on the properties of Pb(Zr(1-x)Ti(x))O3 alloy in its morphotropic phase boundary (MPB) are predicted by combining first principles-based methods and annealing techniques. Clustering is found to lead to a compositional expansion of this boundary, while the association of unlike atoms yields a contraction of this region. Atomic short-range order can thus drastically affect properties of perovskite alloys in their MPB, by inducing phase transitions. Microscopic mechanisms responsible for these effects are revealed and discussed. PMID- 12906675 TI - Fracture mechanisms in bulk metallic glassy materials. AB - We find that the failure of bulk metallic glassy (BMG) materials follows three modes, i.e., shear fracture with a fracture plane significantly deviating from 45 degrees to the loading direction, normal tensile fracture with a fracture plane perpendicular to the loading direction, or distensile fracture in a break or splitting mode with a fracture plane parallel to the loading direction. The actually occurring type of failure strongly depends on the applied loading mode and the microstructure of the material. Extensive evidence indicates that the Tresca fracture criterion is invalid, and for the first time, three fracture criteria are developed for isotropic materials with high strength, such as advanced BMGs or the newly developed bulk nanostructural materials. PMID- 12906676 TI - Bogomol'nyi, Prasad, and Sommerfield configurations in smectics. AB - It is typical in smectic liquid crystals to describe elastic deformations with a linear theory when the elastic strain is small. In smectics, certain essential nonlinearities arise from the requirement of rotational invariance. By employing the Bogomol'nyi, Prasad, and Sommerfield decomposition and relying on boundary conditions and geometric invariants, we have found a large class of exact solutions. We introduce an approximation for the deformation profile far from a spherical inclusion and find an enhanced attractive interaction at long distances due to the nonlinear elasticity, confirmed by numerical minimization. PMID- 12906677 TI - Oxide/metal interface distance and epitaxial strain in the NiO/Ag(001) system. AB - Geometric parameters of NiO films epitaxially grown on Ag(001) were determined using two independent experimental techniques and ab initio simulations. Primary beam diffraction modulated electron emission experiments determined that the NiO films grow with O on top of Ag and that the oxide/metal interface distance is d=2.3+/-0.1 A. Polarization-dependent x-ray absorption, at the Ni-K edge, determined the tetragonal strain (r( parallel )=2.046+/-0.009 A, r( perpendicular )=2.12+/-0.02 A) and d=2.37+/-0.05 A. Periodic slab model results agree with the experiments (d=2.40, r( parallel )=2.07, r( perpendicular )=2.10 A; the O-on-top configuration is the most stable). PMID- 12906678 TI - Bonding of saturated hydrocarbons to metal surfaces. AB - The adsorption of octane on Cu(110) was studied by x-ray absorption and x-ray emission spectroscopy, in combination with spectrum calculations in the framework of density functional theory, as a model system for alkane adsorption on transition metals. Significant electron sharing between the adsorbate and metal surface and involvement of both bonding and antibonding C-H molecular orbitals in the molecule-metal bond was found. The calculations were extended to the case of octane adsorbed on Ni(110), and the position of the metal d band was found to be important for the bonding. The results were generalized to show that this is important for the efficiency as an alkane dehydrogenation catalyst. PMID- 12906679 TI - First-order transition from a Kondo insulator to a ferromagnetic metal in single crystalline FeSi(1-x)Ge(x). AB - The phase diagram of FeSi(1-x)Ge(x), obtained from magnetic, thermal, and transport measurements on single crystals, shows a discontinuous transition from Kondo insulator to ferromagnetic metal with x at a critical concentration, x(c) approximately 0.25. The gap of the insulating phase strongly decreases with x. The specific heat gamma coefficient appears to track the density of states of a Kondo insulator. The phase diagram is consistent with an insulator-metal transition induced by a reduction of the hybridization with x in conjunction with disorder on the Si/Ge ligand site. PMID- 12906680 TI - Optical and loss spectra of carbon nanotubes: depolarization effects and intertube interactions. AB - We performed ab initio calculations of the anisotropic dielectric response of small-diameter single-walled carbon nanotubes in the framework of time-dependent density-functional theory. The calculated optical spectra are in very good agreement with experiment, both concerning absolute peak positions and anisotropy effects. The latter can only be described correctly when crystal local-field effects ("depolarization" effects) are fully taken into account. Moreover, interactions between the tubes can strongly modify their absorption and electron energy-loss spectra. PMID- 12906681 TI - Spin-independent origin of the strongly enhanced effective mass in a dilute 2D electron system. AB - We accurately measure the effective mass in a dilute two-dimensional electron system in silicon by analyzing the temperature dependence of the Shubnikov-de Haas oscillations in the low-temperature limit. A sharp increase of the effective mass with decreasing electron density is observed. We find that the enhanced effective mass is independent of the degree of spin polarization, which points to a spin-independent origin of the mass enhancement and is in contradiction with existing theories. PMID- 12906682 TI - Magnetically induced chessboard pattern in the conductance of a Kondo quantum dot. AB - We quantitatively describe the main features of the magnetically induced conductance modulation of a Kondo quantum dot-or chessboard pattern-in terms of a constant-interaction double quantum dot model. We show that the analogy with a double dot holds down to remarkably low magnetic fields. The analysis is extended by full 3D spin density functional calculations. Introducing an effective Kondo coupling parameter, the chessboard pattern is self-consistently computed as a function of magnetic field and electron number, which enables us to explain our experimental data quantitatively. PMID- 12906683 TI - Zeeman splitting of zero-bias anomaly in Luttinger liquids. AB - Tunneling density of states (DOS) in Luttinger liquid has a dip at zero energy, commonly known as the zero-bias anomaly. In the presence of a magnetic field, in addition to the zero-bias anomaly, the DOS develops two peaks separated from the origin by the Zeeman energy. We show that these finite-bias anomalies are characterized by a power-law behavior of the DOS and the differential conductance, and find the corresponding exponents at arbitrary strength of the electron-electron interaction. The developed theory is applicable to various kinds of quantum wires, including carbon nanotubes. PMID- 12906684 TI - Persistent currents in interacting Aharonov-Bohm interferometers and their enhancement by acoustic radiation. AB - We consider an Aharonov-Bohm interferometer, connected to two electronic reservoirs, with a quantum dot embedded on one of its arms. We find a general expression for the persistent current at steady state, valid for the case where the electronic system is free of interactions except on the dot. The result is used to derive the modification in the persistent current brought about by coupling the quantum dot to a phonon source. The magnitude of the persistent current is found to be enhanced in an appropriate range of the intensity of the acoustic source. PMID- 12906685 TI - Coexistence of composite bosons and composite fermions in nu = 1/2 + 1/2 quantum Hall bilayers. AB - In bilayer quantum Hall systems at filling fractions near nu=1/2+1/2, as the spacing d between the layers is continuously decreased, intralayer correlations must be replaced by interlayer correlations, and the composite fermion (CF) Fermi seas at large d must eventually be replaced by a composite boson (CB) condensate or "111 state" at small d. We propose a scenario where CBs and CFs coexist in two interpenetrating fluids in the transition. Trial wave functions describing these mixed CB-CF states compare very favorably with exact diagonalization results. A Chern-Simons transport theory is constructed that is compatible with experiment. PMID- 12906686 TI - Conductivity of paired composite fermions. AB - We develop a phenomenological description of the nu=5/2 quantum Hall state in which the Halperin-Lee-Read theory of the half-filled Landau level is combined with a p-wave pairing interaction between composite fermions (CFs). The electromagnetic response functions for the resulting mean-field superconducting state of the CFs are calculated and used in an RPA calculation of the q and omega dependent longitudinal conductivity of the physical electrons, a quantity which can be measured experimentally. PMID- 12906687 TI - Chern numbers for spin models of transition metal nanomagnets. AB - We argue that ferromagnetic transition metal nanoparticles with fewer than approximately 100 atoms can be described by an effective Hamiltonian with a single giant spin degree of freedom. The total spin S of the effective Hamiltonian is specified by a Berry curvature Chern number that characterizes the topologically nontrivial dependence of a nanoparticle's many-electron wave function on magnetization orientation. The Berry curvatures and associated Chern numbers have a complex dependence on spin-orbit coupling in the nanoparticle and influence the semiclassical Landau-Liftshitz equations that describe magnetization-orientation dynamics. PMID- 12906688 TI - Deconfinement and dissipation in quantum Hall Josephson tunneling. AB - The zero-bias tunneling resonance in quantum Hall bilayer systems is investigated via numerical simulations of the classical two-dimensional XY model with a symmetry-breaking field. Disorder is included in the model and is shown to nucleate strings of overturned spins proliferated through the system, with unpaired vortices and antivortices at their end points. This string glass state supports low-energy excitations which lead to anomalously large dissipation in tunneling, as observed in experiment. The effect of an in-plane magnetic field is discussed. PMID- 12906689 TI - Influence of pair breaking and phase fluctuations on disordered high Tc cuprate superconductors. AB - Electron irradiation has been used to introduce point defects in a controlled way in underdoped and optimally doped YBa(2)Cu(3)O(7-delta) crystals. This technique allows us to perform very accurate measurements of T(c) and of the ab plane resistivity in a wide range of defect contents x(d) down to T(c)=0. The variation of T(c) and of the transition width with x(d) do not follow current predictions of pair-breaking theories. The data are rather compatible, at least for the highly damaged regime, with the expected influence of phase fluctuations. These results open new questions about the evolution of the defect induced T(c) depression over the phase diagram of the cuprates. PMID- 12906690 TI - Effects of two-band superconductivity on the flux-line lattice in magnesium diboride. AB - We present neutron scattering from the flux line lattice (FLL) in MgB2. Between 0.5 and 0.9 T the FLL undergoes a 30 degrees reorientation, and simultaneously the scattered intensity falls sharply consistent with the weaker superconducting pi band being suppressed with increasing field. We speculate that the pi and sigma bands favor different FLL orientations, and that the reorientation is driven by the suppression of the pi band. When the c axis of the crystal is rotated 45 degrees to the applied field the penetration depth anisotropy could be measured, and rises both as a function of applied field and temperature. PMID- 12906691 TI - Highly degenerate canted spin structure in bilayer manganite La1.1Sr1.9Mn2O7. AB - We have observed magnetic diffuse signals of the bilayer manganite La1.1Sr1.9Mn2O7 by neutron scattering. Even at the low temperature, 20 K, there is a diffuse signal, which is much stronger than that observed in the high temperatures, 144 and 288 K. This signal cannot be characterized by a bilayer structure as observed at 144 K nor by a monolayer structure as at 288 K, but by a high degeneracy of a canted spin structure. Thus, an observation of this diffuse signal is the direct evidence for the establishment of the canted spin structure in La1.1Sr1.9Mn2O7. PMID- 12906692 TI - Magnetization process of single molecule magnets at low temperatures. AB - We show that correlations established before quenching to very low temperatures later drive the magnetization process of systems of single molecule magnets, after a magnetic field is applied at t=0. We also show that in simple cubic lattices m proportional, variant sqrt[t], as observed in Fe8, but only for 1+2log((10)(h(d)/h(w)) time decades, where h(d) is a nearest neighbor dipolar magnetic field and a spin reversal can occur only if the field on it is within ( h(w),h(w)). However, the sqrt[t] behavior is not universal. For bcc and fcc lattices, m proportional, variant t(p), but p approximately 0.7. The value to which m finally levels off is also given. PMID- 12906693 TI - Symmetry of magnetic quantum tunneling in single molecule magnet Mn12-acetate. AB - The symmetry of magnetic quantum tunneling has been studied in the prototype single molecule magnet Mn12-acetate using a micro-Hall effect magnetometer and superconducting high field vector magnet system. An average crystal fourfold symmetry is shown to be due to local molecular environments of twofold symmetry that are rotated by 90 degrees with respect to one another, confirming that disorder which lowers the molecule symmetry is as important to magnetic quantum tunneling. We have studied a subset of these lower (twofold) site symmetry molecules and present evidence for a Berry phase effect consistent with a local twofold symmetry. PMID- 12906694 TI - Field-induced ferromagnetic metallic state of bilayer manganite (La0.4Pr0.6)1.2Sr1.8Mn2O7: a polarized neutron diffraction study. AB - Unpolarized and polarized neutron diffraction measurements have been carried out on the bilayer manganite (La0.4Pr0.6)1.2Sr1.8Mn2O7 which undergoes simultaneous semiconductor-metal paraferromagnetic transitions under magnetic field. Maximum entropy magnetization density reconstruction and multipole refinement on flipping ratios evidence the existence of two distinct field-induced states. The field induced ferromagnetic state where the field is parallel to the c axis is characterized by the presence of magnetic moment on the Sr site of 0.48(2)mu(B), due to the Pr substitution. It also shows a high population of the d3(z(2)-r(2)) orbitals of Mn3+. For the field-induced state where the field is parallel to the a or b axes no magnetization density was found at the Sr site and the dx(2)( y(2)) orbital is slightly more populated than the d3(z(2)-r(2)) one. PMID- 12906695 TI - Entanglement assisted capacity of the broadband Lossy channel. AB - We calculate the entanglement assisted capacity of a multimode bosonic channel with loss. As long as the efficiency of the channel is above 50%, the superdense coding effect can be used to transmit more bits than those that can be stored in the message sent down the channel. Bounds for the other capacities of the multimode channel are also provided. PMID- 12906696 TI - Imprinting complete information about a quantum channel on its output state. AB - We introduce a novel property of bipartite quantum states, which we call faithfulness, and we say that a state is faithful when acting with a channel on one of the two quantum systems; the output state carries complete information about the channel. The concept of faithfulness can also be extended to sets of states, when the output states altogether carry a complete imprinting of the channel. Measures of degrees of faithfulness are proposed. PMID- 12906697 TI - Combinatorics and quantum nonlocality. AB - We use techniques for lower bounds on communication to derive necessary conditions (in terms of detector efficiency or amount of superluminal communication) for being able to reproduce the quantum correlations occurring in Einstein-Podolsky-Rosen-type experiments with classical local hidden-variable theories. As an application, we consider n parties sharing a Greenberger-Horne Zeilinger-type state and show that the amount of superluminal classical communication required to reproduce the correlations is at least n(log((2)n-3) bits and the maximum detector efficiency eta(*) for which the resulting correlations can still be reproduced by a local hidden-variable theory is upper bounded by eta(*)99%) fragment in a nonergodic process. In solution at natural pH the AMP anion is prevalent and photoproduct formation is therefore less important as the time for vibrational cooling is of the order of picoseconds. PMID- 12906700 TI - Particle dispersion on rapidly folding random heteropolymers. AB - We investigate the dynamics of a particle moving randomly along a disordered heteropolymer subjected to rapid conformational changes which induce superdiffusive motion in chemical coordinates. We study the antagonistic interplay between the enhanced diffusion and the quenched disorder. The dispersion speed exhibits universal behavior independent of the folding statistics. On the other hand it is strongly affected by the structure of the disordered potential. The results may serve as a reference point for a number of translocation phenomena observed in biological cells, such as protein dynamics on DNA strands. PMID- 12906701 TI - Comment on "Locality and topology in the molecular Aharonov-Bohm effect". PMID- 12906703 TI - Comment on "ac conductivity spectra of alkali tellurite glasses: composition dependent deviations from the Summerfield scaling". PMID- 12906705 TI - Comment on "Time-dependent density-matrix renormalization group: a systematic method for the study of quantum many-body out-of-equilibrium systems". PMID- 12906707 TI - Comment on "Motional ordering of a charge-density wave in the sliding state". PMID- 12906709 TI - Calpain cleavage of the B isoform of Ins(1,4,5)P3 3-kinase separates the catalytic domain from the membrane anchoring domain. AB - Inositol (1,4,5)-trisphosphate [Ins(1,4,5)P3] is one of the key intracellular second messengers in cells and mobilizes Ca2+ stores in the ER (endoplasmic reticulum). Ins(1,4,5)P3 has a short half-life within the cell, and is rapidly metabolized through one of two pathways, one of which involves further phosphorylation of the inositol ring: Ins(1,4,5)P3 3-kinase (IP3-3K) phosphorylates Ins(1,4,5)P3, resulting in the formation of inositol (1,3,4,5) tetrakisphosphate [Ins(1,3,4,5)P4]. There are three known isoforms of IP3-3K, designated IP3-3KA, IP3-3KB and IP3-3KC. These have differing N-termini, but highly conserved C-termini harbouring the catalytic domain. The three IP3-3K isoforms have different subcellular locations and the B-kinase is uniquely present in both cytosolic and membrane-bound pools. As it is the N-terminus of the B-kinase that differs most from the A- and C-kinases, we have hypothesized that this portion of the protein may be responsible for membrane localization. Although there are no known membrane-targeting protein motifs within the sequence of IP3-3KB, it is found to be tightly associated with the ER membrane. Here, we show that specific regions of the N-terminus of IP3-3KB are necessary and sufficient for efficient membrane localization of the protein. We also report that, in the presence of Ca2+, the kinase domain of IP3-3KB is cleaved from the membrane-anchoring region by calpain. PMID- 12906710 TI - Interleukin-1-receptor-associated kinase 2 (IRAK2)-mediated interleukin-1 dependent nuclear factor kappaB transactivation in Saos2 cells requires the Akt/protein kinase B kinase. AB - The post-receptor pathway that leads to nuclear factor kappaB (NF-kappaB) activation begins with the assembly of a membrane-proximal complex among the interleukin 1 (IL-1) receptors and the adaptor molecules, myeloid differentiation protein 88 (MyD88), IL-1-receptor-associated kinases (IRAKs) and tumour-necrosis factor-receptor-associated factor 6. Eventually, phosphorylation of the inhibitor of NF-kappaB (IkappaB) by the IkappaB kinases releases NF-kappaB, which translocates to the nucleus and modulates gene expression. In this paper, we report that IRAK2 and MyD88, but not IRAK1, interact physically with Akt, as demonstrated by co-immunoprecipitation and pull-down experiments. Interestingly, the association of Akt with recombinant IRAK2 is decreased by stimulation with IL 1, and is favoured by pre-treatment with phosphatase. Likewise, Akt association with IRAK2 is increased considerably by overexpression of PTEN (phosphatase and tensin homologue deleted on chromosome 10), while it is completely abrogated by overexpression of phosphoinositide-dependent protein kinase 1. These data indicate that Akt takes part in the formation of the signalling complex that conveys the signal from the IL-1 receptors to NF-kappaB, a step that is much more membrane-proximal than was reported previously. We also demonstrate that Akt activity is necessary for IL-1-dependent NF-kappaB transactivation, since a kinase-defective mutant of Akt impairs IRAK2- and MyD88-dependent, but not IRAK1 dependent, NF-kappaB activity, as monitored by a gene reporter assay. Accordingly, IRAK2 failed to trigger inducible nitric oxide synthase and IL-1beta production in cells expressing dominant-negative Akt. However, NF-kappaB binding to DNA was not affected by inhibition of Akt, indicating that Akt regulates NF kappaB at a level distinct from the dissociation of p65 from IkappaBalpha and its translocation to the nucleus, possibly involving phosphorylation of the p65 transactivation domain. PMID- 12906711 TI - Incorporation of whey proteins into cheese curd by using transglutaminase. AB - A Ca(2+)-independent microbial TGase (transglutaminase) isolated from Streptoverticillium mobaraense was used to obtain whey protein containing novel dairy products. We evaluated the difference both in the curd formation time as well as in the hardness and deformability of the cheese obtained from cow's milk in the presence or absence of the enzyme. The results of our experiments showed that the milk coagulation time was dependent on the step in cheese manufacture at which TGase was added. We analysed the deformability and the hardness of the dairy products obtained either by adding both TGase and the milk-clotting enzyme to the milk sample at the same time or by adding TGase after treating the milk sample for 30 min with the clotting enzyme and cutting the obtained coagulum. TGase treatment conferred a strongly decreased protein content to derived whey. Moreover, when further amounts of whey were added to the milk during the manufacturing process in the presence of TGase, whey-protein-enriched dairy products could also be obtained. Our findings may lead to new biotechnologies for the re-utilization of by-products from dairy plants and contribute to reduction of environmental pollution from whey-protein disposal. PMID- 12906712 TI - Monochorionic-triamniotic triplet pregnancy after intracytoplasmic sperm injection, assisted hatching, and two-embryo transfer: first reported case following IVF. AB - BACKGROUND: We present a case of monochorionic-triamniotic pregnancy that developed after embryo transfer following in vitro fertilization (IVF). METHODS: After controlled ovarian hyperstimulation and transvaginal retrieval of 22 metaphase II oocytes, fertilization was accomplished with intracytoplasmic sperm injection (ICSI). Assisted embryo hatching was performed, and two embryos were transferred in utero. One non-transferred blastocyst was cryopreserved. RESULTS: Fourteen days post-transfer, serum hCG level was 423 mIU/ml and subsequent transvaginal ultrasound revealed a single intrauterine gestational sac with three separate amnion compartments. Three distinct foci of cardiac motion were detected and the diagnosis was revised to monochorionic-triamniotic triplet pregnancy. Antenatal management included cerclage placement at 19 weeks gestation and hospital admission at 28 weeks gestation due to mild preeclampsia. Three viable female infants were delivered via cesarean at 30 5/7 weeks gestation. CONCLUSIONS: The incidence of triplet delivery in humans is approximately 1:6400, and such pregnancies are classified as high-risk for reasons described in this report. We also outline an obstetric management strategy designed to optimize outcomes. The roles of IVF, ICSI, assisted embryo hatching and associated laboratory culture conditions on the subsequent development of monozygotic/monochorionic pregnancy remain controversial. As demonstrated here, even when two-embryo transfer is employed after IVF the statistical probability of monozygotic multiple gestation cannot be reduced to zero. We encourage discussion of this possibility during informed consent for the advanced reproductive technologies. PMID- 12906713 TI - Over-expression of the mitogen-activated protein kinase (MAPK) kinase (MEK)-MAPK in hepatocellular carcinoma: its role in tumor progression and apoptosis. AB - BACKGROUND: Hepatocellular carcinoma (HCC) is one of the most common malignancies in South East Asia. Although activation of the MEK-MAPK is often associated with cellular growth, the role of MEK-MAPK in growth and survival of hepatocarcinoma cells has not been established. METHODS: Immuno-histochemistry was used to localize phosphorylated MAPK and MEK1/2 in the tissues. 3-(4,5-dimethylthiazol-2 y1)-2,5-diphenyltetrazolium bromide (MTT) assay and ELISA were used to determine cell viability and cell proliferation. Deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) assay was used to detect apoptotic cells. Western blots analysis was performed to determine the levels of proteins involved in the MEK MAPK and apoptotic pathways. Transfection study was performed to assess the role of MEK-MAPK pathway in growth and survival of liver cancer cells. RESULTS: We report that phosphorylation of MEK1/2 at Ser217/221 was detected by immuno histochemistry in 100% (46 of 46) of HCCs examined. A positive signal was localized in the nuclei of hepatocarcinoma cells but not in dysplastic hepatocytes or stromal cells. Over-expression and phosphorylation of MAPK was also detected in 91% (42 of 46) and 69% (32 of 46) of HCCs examined, respectively. The percentage of cells showing positively for phosphorylated MEK1/2 increased with advancing tumor stage. In vitro, treatment of human HepG2 and Hep3B cells with MEK1/2 specific inhibitors U0126 and PD98059 led to growth inhibition and apoptosis. U0126 induced the release of cytochrome c and increased the cleavage of caspase-3, caspase-7, and poly ADP-ribose polymerase (PARP). Inhibition of phosphatidylinositol 3-kinase (PI-3K), c-Jun N-terminal kinase (JNK) and p38 kinase activities caused only a mild apoptosis in HepG2 and Hep3B cells. Activated MEK1-transfected cells were more resistant to UO126-induced apoptosis in vitro and formed larger tumors in SCID mice than mock-transfected cells. CONCLUSION: In conclusion, our results demonstrate that MEK-MAPK plays an important role in the growth and survival of liver cancer cells and suggest that blocking MEK-MAPK activity may represent an alternative approach for the treatment of liver cancer. PMID- 12906714 TI - A case of sigmoid endometriosis difficult to differentiate from colon cancer. AB - BACKGROUND: Although endometriosis with sigmoid serosal involvement is not uncommon in women of childbearing age, the mucosal involvement is rare and differential diagnosis from colon cancer may be difficult due to the lack of pathognomonic symptoms and the poor diagnostic yield of colonoscopy and colonic biopsies. CASE PRESENTATION: We present a case of a young woman with sigmoid endometriosis, in which the initial diagnostic workup suggested colon cancer. Histologic evidence, obtained from a second colonoscopy, along with pelvic ultrasound findings led to the final diagnosis of intestinal endometriosis which was confirmed by laparoscopy. CONCLUSION: Colonic endometriosis is often a diagnostic challenge and should be considered in young women with symptoms from the lower gastrointestinal tract. PMID- 12906715 TI - Developmental gene regulation during tomato fruit ripening and in-vitro sepal morphogenesis. AB - BACKGROUND: Red ripe tomatoes are the result of numerous physiological changes controlled by hormonal and developmental signals, causing maturation or differentiation of various fruit tissues simultaneously. These physiological changes affect visual, textural, flavor, and aroma characteristics, making the fruit more appealing to potential consumers for seed dispersal. Developmental regulation of tomato fruit ripening has, until recently, been lacking in rigorous investigation. We previously indicated the presence of up-regulated transcription factors in ripening tomato fruit by data mining in TIGR Tomato Gene Index. In our in-vitro system, green tomato sepals cultured at 16 to 22 degrees C turn red and swell like ripening tomato fruit while those at 28 degrees C remain green. RESULTS: Here, we have further examined regulation of putative developmental genes possibly involved in tomato fruit ripening and development. Using molecular biological methods, we have determined the relative abundance of various transcripts of genes during in vitro sepal ripening and in tomato fruit pericarp at three stages of development. A number of transcripts show similar expression in fruits to RIN and PSY1, ripening-associated genes, and others show quite different expression. CONCLUSIONS: Our investigation has resulted in confirmation of some of our previous database mining results and has revealed differences in gene expression that may be important for tomato cultivar variation. We present new and intriguing information on genes that should now be studied in a more focused fashion. PMID- 12906716 TI - Influence of the TNF-alpha and TNF-beta polymorphisms upon infectious risk and outcome in surgical intensive care patients. AB - BACKGROUND: Tumor necrosis factor-alpha (TNF-alpha) is a well-documented central inflammatory mediator in sepsis. Specific polymorphisms of the TNF-alpha and TNF beta genes (TNF2 and LTA + 250, respectively) have been suggested to correlate with higher mortality in septic shock. This study sought to determine whether these polymorphisms of the TNF-alpha and -beta genes are associated with an increased risk of infection in an at-risk surgical intensive care population. MATERIALS AND METHODS: Forty-four consecutive patients with systemic inflammatory response syndrome were enrolled prospectively in the study. Genomic DNA was isolated from whole blood samples using standard phenol/chloroform extraction techniques. Specific fragments including the polymorphic sites of each gene were amplified by polymerase chain reaction, and restriction enzyme digestions were performed. Genotypes were determined by gel electrophoresis and confirmed by direct sequencing. RESULTS: Eighty-six percent of the patients were TNF1 homozygotes (G:G at -308 of the TNF-alpha promoter region), whereas 9% of the patients were homozygous for TNF2 (A:A). There was no difference in the incidence of sepsis, septic shock, or mortality between patients bearing the various alleles. Only 13.6% of the patients exhibited the G:G alleles for TNF-beta, whereas the homozygous A:A was present in 45.4% of the patients. CONCLUSION: The presence of the A allele at these polymorphic sites did not predispose critically ill surgical patients to either infection or septic shock. PMID- 12906717 TI - Lower levels of whole blood LPS-stimulated cytokine release are associated with poorer clinical outcomes in surgical ICU patients. AB - BACKGROUND: In vitro pretreatment of human monocytes with lipopolysaccharide (LPS) induces "endotoxin tolerance" with blunted TNF and IL-6 release to rechallenge with LPS. The pro-inflammatory cytokines TNF and IL-6 are important mediators in sepsis. A high IL-6 concentration has been used as a marker of infection severity, but IL-6 may also have beneficial effects as an acute-phase protein. We sought to address two questions: (a) What is the relationship between TNF and IL-6 release? (b) Is the clinical outcome different for intensive care unit (ICU) patients with ex vivo characteristics of endotoxin tolerance (low levels of ex vivo LPS-stimulated cytokine release)? MATERIALS AND METHODS: Heparinized whole blood was obtained from 62 surgical ICU patients and 15 control subjects and incubated for 3 h at 37 degrees C in the presence or absence of 10 ng/mL LPS. Concentrations of TNF and IL-6 were measured in plasma samples using an enzyme-linked immunosorbent assay (pg/mL). Clinical data on ICU length of stay (LOS), ventilator days, white blood cell count (WBC), and documented clinical infection were obtained by chart review. Outcome parameters for patients with low ex vivo LPS-stimulated cytokine release (low = IL-6 < 3000 pg/mL and TNF <2100 pg/mL) were compared to patients with Normal/High concentrations of cytokines. RESULTS: Cytokines were essentially undetectable in ICU patients or controls without LPS stimulation, however a range of values was measured for LPS stimulated release in both ICU patients (IL-6, 7847 +/- 857 pg/mL; TNF, 4390 +/- 457 pg/mL) and controls (IL-6, 7704 +/- 793 pg/mL; TNF, 6706 +/- 715 pg/mL). There were no differences in age between High/Normal concentrations of cytokines compared to the Low cytokine group, however there were significant differences in WBC, cytokine concentrations, ICU LOS, incidence of clinical infection, and mortality. The Low group also required an average of 6.9 more days of mechanical ventilation (p < 0.05). LPS-stimulated TNF release seemed to correlate better with the observed mortality than did IL-6 release. CONCLUSION: The data suggest that ICU patients with characteristics of endotoxin tolerance (low LPS-stimulated cytokine release capacity) have significantly poorer clinical outcomes. Ex vivo LPS-stimulated whole blood cytokine production may be useful to identify ICU patients with severe sepsis. PMID- 12906718 TI - Lipoteichoic acid is a potent inducer of cytokine production in rat and human Kupffer cells in vitro. AB - BACKGROUND: Kupffer cells have been proposed to be a major cellular origin of pro inflammatory mediators in sepsis. However, the cytokine response of Kupffer cells to gram-positive bacteria and their endotoxins peptidoglycan (PepG) and lipoteichoic acid (LTA) has never previously been studied. MATERIALS AND METHODS: Primary cultures of rat and human Kupffer cells were exposed to live Staphylococcus aureus (S. aureus) (4.0 x 10(1) to 4.0 x 10(7) CFU/mL culture medium), as well as highly purified PepG and LTA (0-100 microg/mL). Lipopolysaccharide (LPS) at 1 microg/mL was used for control. In parallel experiments, whole blood obtained from the same rats was stimulated in a similar manner. Accumulation of the proinflammatory cytokines tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) in plasma or culture supernatants were assessed by enzyme immuno assays. TNF-alpha and IL-6 mRNA were analyzed by real time RT-PCR. RESULTS: PepG and LTA, as well as live S. aureus, induced the production of TNF-alpha and IL-6 in Kupffer cells from both species in a time- and dose-dependent manner. Whereas PepG was a more potent inducer of TNF-alpha and IL-6 in whole blood, the opposite seemed to be the case in Kupffer cells. In fact, a 100-fold lower concentration of LTA (1 microg/mL) than of PepG (100 microg/mL) was sufficient to induce a substantial production of both TNF-alpha and IL-6 in the Kupffer cells. TNF-alpha and IL-6 mRNA were induced correspondingly. CONCLUSION: Our results support the contention that gram positive bacteria may activate cytokine production in Kupffer cells during bacteremia and suggest that LTA is important in this interaction. PMID- 12906719 TI - Serum bicarbonate concentration correlates with arterial base deficit in critically ill patients. AB - BACKGROUND: Base deficit (BD) and lactate concentration have been established as endpoints of resuscitation (EOR) in critically ill patients. However, obtaining these data has traditionally required an arterial blood gas (ABG) sample. We hypothesized that the more easily available serum bicarbonate (SB) concentration could approximate BD and potentially serve as a useful EOR of critically ill or septic patients. We evaluated retrospectively the correlation of SB with BD in a cohort of surgical intensive care unit patients. MATERIALS AND METHODS: Clinical data from April 1996 through April 1998 were recorded in a computerized application from 1,712 critically ill adult patients. The data were downloaded daily and imported into a relational database for storage and analysis. A subset of paired SB and ABG samples obtained simultaneously was analyzed by linear regression to determine the correlation coefficients (r) and coefficient of determinations (r(2)) for the respective analyses. RESULTS: A total of 26,690 BD and 16,737 SB determinations were available in the database. Of these, 5,301 BD and SB samples were drawn simultaneously on the same patient. The correlation coefficient for these data pairs was 0.91, and the coefficient of determination was 0.83. The base deficit was predicted by the equation: BD = 22.43 - (0.9522 x SB) (p < 0.0001). CONCLUSION: In this large data set, there was a close inverse correlation between SB and BD in critically ill or septic patients. The predictive equation explains 83% of the variability for BD values. A prospective study comparing SB to BD and lactate could confirm SB as a useful marker of resuscitation. PMID- 12906720 TI - Fungal sinusitis in the immunocompetent patient: risk factors and surgical management. AB - BACKGROUND: Fungal sinusitis has been reported increasingly in immunocompetent patients. However, the most effective, appropriate mode of therapy has not been determined. MATERIALS AND METHODS: In this retrospective study, we examined the records of 110 immunocompetent patients with chronic sinusitis who had undergone sinus surgery at our institution between 1983 and 1994. Five patients (4.5%) with fungal sinusitis were identified. Information on those patients was compared with that of the 105 patients with nonfungal sinusitis. RESULTS: Prolonged use of topical steroids was no more common in patients with fungal sinusitis (20%) than it was in patients with nonfungal sinusitis (4.8%) (p = 0.25). Differentiating features of fungal sinusitis were the presence of a metallic density and areas of high- and low-density on radiologic examination (p < 0.01). All five of the patients with fungal sinusitis were cured by surgical intervention, primarily endoscopic sinus surgery, without undergoing anti-mycotic therapy. Four of the five patients were followed up for at least 3 years without any recurrence. CONCLUSION: Endoscopic sinus surgery is and should remain the mainstay of treatment for fungal sinusitis in immunocompetent patients. Adjunctive anti mycotic therapy may not be necessary. PMID- 12906721 TI - Surgical treatment of severe acute pancreatitis: timing of operation is crucial for survival. AB - BACKGROUND: In patients operated on for severe acute pancreatitis (SAP) the impact of the timing of operation on outcome is controversial. MATERIALS AND METHODS: In a retrospective analysis of a prospectively documented database, we studied 250 patients suffering from SAP, who were in need for surgical treatment during their course of disease. RESULTS: From 1982 to 1998, 250 patients with the diagnosis of SAP who required operative treatment were admitted to the intensive care unit (ICU) of a university hospital. The mean APACHE II score on the day of admission was 16.1 (8-35). One hundred eighty-five patients (74%) required reoperation, of whom 111 patients (60%) underwent reoperation on demand and 74 (40%) patients a pre-planned reoperation. Overall mortality was 38.8% (97 patients). In patients who were operated during the first three weeks after onset of disease, mortality was significantly higher than in patients who were operated after three weeks (46% vs. 25%, p < 0.01). Besides patient age (p < 0.05), APACHE II score at admission (p < 0.01), multiple organ dysfunction (p < 0.01), infection of pancreatic necrosis (p < 0.05), surgical control of pancreatic necrosis (p < 0.0001), and the time of surgical intervention (p < 0.05) determined survival significantly. CONCLUSION: Patients who were operated later than three weeks after onset of disease had a significantly better outcome. In patients suffering from SAP who required surgical treatment, the timing of operation is crucial for survival. PMID- 12906722 TI - Appendicitis: selective use of abdominal CT reduces negative appendectomy rate. AB - BACKGROUND: Recent literature supports the use of CT for all patients with right lower quadrant pain to improve diagnostic accuracy and reduce hospital cost. Our current practice at a large teaching hospital is to use CT only for patients whose diagnosis remains in question after a thorough history, physical examination, and focused laboratory assessment. In consideration of a more liberal use of CT for right lower quadrant pain, we reviewed our experience with selective CT and the effect this has had on our practice, paying particular attention to perforation rate and negative appendectomy rate. MATERIALS AND METHODS: A retrospective chart review was performed for all patients who underwent appendectomy in 1995 and 1998. These two patient populations were evaluated for patient demographics, use of CT scan, perforation rate, and incidence of negative appendectomy. RESULTS: Three hundred ninety-four patients underwent appendectomy in 1995 versus 372 in 1998. Gender (64% male in 1995 vs. 60% in 1998), age (mean 24.9 vs. 25.5) and perforation rate (31% in 1995 vs. 29% in 1998) did not change. The use of CT increased in 1998 (12% vs. 34%, p < 0.001) and overall nontherapeutic appendectomy decreased (14% vs. 7%, p < 0.005). This decrease was seen in patients who were operated without CT (13% vs. 7%, p < 0.03) and after CT (19% vs. 6%, p < 0.02). CONCLUSION: Selective use of CT scan in patients presenting with right lower quadrant pain is helpful in reducing the incidence of nontherapeutic appendectomy. PMID- 12906723 TI - Honey in the management of infections. AB - BACKGROUND: Honey, a natural product of bees of the genera Apis and Meliponinae, has been recognized for medicinal properties since antiquity. Honey has demonstrated antimicrobial properties. These effects are variably ascribed to the pH, hydrogen peroxide content, osmotic effect, and as yet unidentified compounds putatively described as inhibines. MATERIALS AND METHODS: This review will explore the use of honey in necrotizing soft tissue infections, postsurgical wound infections, wounds other than postsurgical infections, Helicobacter pylori of the stomach and duodenum, and burns. Throughout, the in vitro evidence that exists and the explanations that can be offered for the purported benefits of honey will be reviewed. Most of the reports are either uncontrolled case series or in vitro observations. As such, detailed critique of statistical methods will not be undertaken. CONCLUSION: The purpose of this paper is not to debunk honey therapy as a myth, but to stimulate thought among surgeons interested in surgical infection and perhaps serve as the nidus for future research. The use of honey should be considered when more conventional therapies have failed. PMID- 12906724 TI - A rare manifestation of perforated diverticulitis: parastomal subcutaneous abscess. AB - Perforation is a serious complication of diverticular disease. The sigmoid is the main affected anatomic site of perforated diverticulitis and sigmoid resection followed either by Hartmann procedure or primary anastomosis are the standard surgical approaches. Surgery, however, does not cure diverticular disease. About 50% of patients have residual diverticula. The morbidity of residual diverticula appears to be low [1]. However, adequate follow-up studies on recurrent diverticulitis after surgery are lacking. We report a rare case of recurrent perforated diverticulitis in the subcutaneous tissue in proximity of the colostomy after a Hartmann procedure for perforated diverticulitis of the sigmoid colon. PMID- 12906725 TI - A taxonomy for artificial embryogeny. AB - A major challenge for evolutionary computation is to evolve phenotypes such as neural networks, sensory systems, or motor controllers at the same level of complexity as found in biological organisms. In order to meet this challenge, many researchers are proposing indirect encodings, that is, evolutionary mechanisms where the same genes are used multiple times in the process of building a phenotype. Such gene reuse allows compact representations of very complex phenotypes. Development is a natural choice for implementing indirect encodings, if only because nature itself uses this very process. Motivated by the development of embryos in nature, we define artificial embryogeny (AE) as the subdiscipline of evolutionary computation (EC) in which phenotypes undergo a developmental phase. An increasing number of AE systems are currently being developed, and a need has arisen for a principled approach to comparing and contrasting, and ultimately building, such systems. Thus, in this paper, we develop a principled taxonomy for AE. This taxonomy provides a unified context for long-term research in AE, so that implementation decisions can be compared and contrasted along known dimensions in the design space of embryogenic systems. It also allows predicting how the settings of various AE parameters affect the capacity to efficiently evolve complex phenotypes. PMID- 12906726 TI - Evolving small neurocontrollers with self-organized compact encoding. AB - This article presents a novel method for the evolution of artificial autonomous agents with small neurocontrollers. It is based on adaptive, self-organized compact genotypic encoding (SOCE) generating the phenotypic synaptic weights of the agent's neurocontroller. SOCE implements a parallel evolutionary search for neurocontroller solutions in a dynamically varying and reduced subspace of the original synaptic space. It leads to the emergence of compact successful neurocontrollers starting from large networks. The method can serve to estimate the network size needed to perform a given task, and to delineate the relative importance of the neurons composing the agent's controller network. PMID- 12906727 TI - Several necessary conditions for the evolution of complex forms of life in an artificial environment. AB - In order for an artificial life (Alife) system to evolve complex creatures, an artificial environment prepared by a designer has to satisfy several conditions. To clarify this requirement, we first assume that an artificial environment implemented in the computational medium is composed of an information space in which elementary symbols move around and react with each other according to human prepared elementary rules. As fundamental properties of these factors (space, symbols, transportation, and reaction), we present ten criteria from a comparison with the biochemical reaction space in the real world. Then, in the latter half of the article, we take several computational Alife systems one by one, and assess them in terms of the proposed criteria. The assessment can be used not only for improving previous Alife systems but also for devising new Alife models in which complex forms of artificial creatures can be expected to evolve. PMID- 12906728 TI - Intelligent meaning creation in a clumpy world helps communication. AB - This article investigates the problem of how language learners decipher what words mean. In many recent models of language evolution, agents are provided with innate meanings a priori and explicitly transfer them to each other as part of the communication process. By contrast, I investigate how successful communication systems can emerge without innate or transferable meanings, and show that this is dependent on the agents developing highly synchronized conceptual systems. I present experiments with various cognitive, communicative, and environmental factors which affect the likelihood of agents achieving meaning synchronization and demonstrate that an intelligent meaning creation strategy in a clumpy world leads to the highest level of meaning similarity between agents. PMID- 12906729 TI - Evolutionary plantographics. AB - This letter describes an evolutionary system for creating lifelike three dimensional plants and flowers, our main goal being the facilitation of producing realistic plant imagery. With these two goals in mind--ease of generation and realism--we designed the plant genotype and the genotype-to-phenotype mapping. Diversity in our system comes about through two distinct processes--evolution and randomization--allowing the creation not only of single plants but of entire gardens and forests. Thus, we are able to readily produce natural-looking artificial scenes. PMID- 12906730 TI - Collective intelligence of the artificial life community on its own successes, failures, and future. AB - We describe a novel Internet-based method for building consensus and clarifying conflicts in large stakeholder groups facing complex issues, and we use the method to survey and map the scientific and organizational perspectives of the artificial life community during the Seventh International Conference on Artificial Life (summer 2000). The issues addressed in this survey included artificial life's main successes, main failures, main open scientific questions, and main strategies for the future, as well as the benefits and pitfalls of creating a professional society for artificial life. By illuminating the artificial life community's collective perspective on these issues, this survey illustrates the value of such methods of harnessing the collective intelligence of large stakeholder groups. PMID- 12906731 TI - Molecular and immunological approaches to controlling parasitic infection. PMID- 12906732 TI - The design and analysis of microarray experiments: applications in parasitology. AB - Microarray experiments can generate enormous amounts of data, but large datasets are usually inherently complex, and the relevant information they contain can be difficult to extract. For the practicing biologist, we provide an overview of what we believe to be the most important issues that need to be addressed when dealing with microarray data. In a microarray experiment we are simply trying to identify which genes are the most "interesting" in terms of our experimental question, and these will usually be those that are either overexpressed or underexpressed (upregulated or downregulated) under the experimental conditions. Analysis of the data to find these genes involves first preprocessing of the raw data for quality control, including filtering of the data (e.g., detection of outlying values) followed by standardization of the data (i.e., making the data uniformly comparable throughout the dataset). This is followed by the formal quantitative analysis of the data, which will involve either statistical hypothesis testing or multivariate pattern recognition. Statistical hypothesis testing is the usual approach to "class comparison," where several experimental groups are being directly compared. The best approach to this problem is to use analysis of variance, although issues related to multiple hypothesis testing and probability estimation still need to be evaluated. Pattern recognition can involve "class prediction," for which a range of supervised multivariate techniques are available, or "class discovery," for which an even broader range of unsupervised multivariate techniques have been developed. Each technique has its own limitations, which need to be kept in mind when making a choice from among them. To put these ideas in context, we provide a detailed examination of two specific examples of the analysis of microarray data, both from parasitology, covering many of the most important points raised. PMID- 12906733 TI - Genomics and its impact on parasitology and the potential for development of new parasite control methods. AB - Parasitic organisms remain the scourge of the developed and underdeveloped worlds. Malaria, schistosomiasis, leishmaniasis, and trypanosomiasis, for example, still result in a large number of human deaths each year worldwide, while drug resistance among nematodes still poses a major problem to the livestock industries. Genome projects involving parasitic organisms are now abundant, and technologies for the investigations of the parasite transcriptome and proteome are well established. There is no doubt the era of the "omics" is with parasitology, and current trends in the discipline are addressing fundamental biological questions that can make best use of the new technologies, as well as the vast amount of new data being generated. Will this become the "golden age of molecular parasitology," leading to the control of parasitic diseases that have plagued mankind for hundreds of years? The primary aim of this paper is to review advances in the general area of parasite genomics, and to outline where the application of "omics" technologies can and have impacted on the development of new control methods for parasitic organisms. PMID- 12906734 TI - Parasites and immune responses: memory illusion? AB - Immunological memory responses to intracellular protozoa and extracellular helminths govern host resistance and susceptibility to reinfection. Humans and livestock living in parasitic disease endemic regions face continuous exposure from a very early age that often leads to asymptomatic chronic infection over their entire lifespan. Fundamental immunological studies suggest that the generation of T-cell memory is driven by tightly coordinated innate and adaptive cellular immune responses rapidly triggered following initial host infection. A key distinguishing feature of immune memory maintenance between the majority of parasitic diseases and most bacterial or viral diseases is long-term antigen persistence. Consequently, functional parasite immune memory is in a continuous, dynamic flux between activation and deactivation producing functional parasite killing or functional memory cell death. In this sense, T-cell immune memory can be regarded as "memory illusion." Furthermore, due to the finite capacity of memory lymphocytes to proliferate, continuous parasite antigen stimulation may exceed a threshold level at some point in the chronically infected host. This may result in suboptimal effector immune memory leading to host susceptibility to reinfection, or immune dysregulation yielding disease reactivation or immune pathology. The goal of this review is to highlight, through numerous examples, what is currently known about T-cell immune memory to parasites and to provide compelling hypotheses on the survival and maintenance of parasite "memory illusion." These novel concepts are discussed in the context of rationale parasite vaccine design strategies. PMID- 12906735 TI - Parasitic infection and the polarized Th2 immune response can alter a vaccine induced immune response. AB - The AIDS epidemic in the Developing World represents a major global crisis. It is imperative that we develop an effective vaccine. Vaccines are economically the most efficient means of controlling viral infections. However, the development of a vaccine against HIV-1 has been a formidable task, and in developing countries chronic parasitic infection adds another level of complexity to AIDS vaccine development. Helminthic and protozoan infections, common in developing countries, can result in a constant state of immune activation that is characterized by a dominant Th2 type of cytokine profile, high IgE levels, and eosinophilia. Such an immune profile may have an adverse impact on the efficacy of vaccines, in particular, an HIV-1 vaccine. Indeed, the CD8 cellular immune response and the corresponding Th1 type cytokines that enhance the CD8 cellular immune response are important for clearing many viral infections. It is believed that an antigen specific CD8 cellular immune response will be an important component of an HIV-1 vaccine. PMID- 12906736 TI - The role of the host immune response in pathogenesis of Bartonella henselae. AB - Bartonella henselae can infect humans resulting in a wide range of disease syndromes including cat-scratch disease, fever with bacteremia, endocarditis, bacillary angiomatosis, and bacillary peliosis hepatis, among others. The nature and severity of the clinical presentation correlates well with the status of the hosts' immune system. Individuals with impaired immune function, including HIV infection, progress to systemic infections more often. Patients with intact immune function who become infected with B. henselae usually get cat-scratch disease, a disease that usually involves lymphadenopathy resulting from a strong cellular immune response to the bacterium. However, immunocompromised patients often progress to bacillary angiomatosis or bacillary peliosis hepatis. The reduced ability of the hosts immune response to control bacterial infection apparently results in a bacteremia of longer duration, and in some patients the presence of angiogenic lesions that are unique among bacterial infections to Bartonella. Recently, the role of immune effector cells that produce angiogenic cytokines upon stimulation with B. henselae has been proposed. Here, the current status of the role of the immune response in both controlling infection and in B. henselae-triggered immunopathogenesis is presented. PMID- 12906737 TI - Traumatic brain injury induces nociceptin/orphanin FQ expression in neurons of the rat cerebral cortex. AB - Nociceptin/orphanin FQ (N/OFQ) is a recently identified opioid-related neuropeptide. Earlier in vitro studies revealed regulation of N/OFQ expression by injury-induced factors, such as ciliary neurotrophic factor, inflammatory cytokines, and reactive oxygen species. We have extended our studies to in vivo experiments investigating the effect of traumatic brain injury on N/OFQ gene expression and peptide levels in the rat brain. Stab wound injury to the rat cerebral cortex led to a significant increase in N/OFQ mRNA levels in the vicinity of the injury, with the largest induction being seen at 24 h post injury. Quantitative in situ hybridization revealed an almost twofold increase in the number of cells expressing N/OFQ, and the signal intensities within cells were also elevated. Stab wound injury leads to proliferation and hypertrophy of astrocytes, which respond to injury-related factors in vitro by up-regulating N/OFQ expression. However, in vivo N/OFQ co-localized exclusively with the neuronal marker, NeuN, following injury. N/OFQ expression was not detected in caspase-3-positive neurons undergoing apoptosis following injury, and increased N/OFQ expression was spatially more extended than the secondary injury-induced responses, such as astrogliosis and neuronal degeneration. Elevation of N/OFQ immunoreactivity closely followed the increase in N/OFQ gene expression as determined by immunohistochemistry. N/OFQ selectively activates the NOP receptor (ORL-1), but we did not detect parallel changes in levels of NOP receptor mRNA following injury, indicating regulation of the nociceptin system at the peptide and not the receptor level. In summary, a profound and prolonged up-regulation of N/OFQ expression in neurons surrounding a stab wound lesion to cerebral cortex was detected. The function of N/OFQ up-regulation in injury-induced responses in the brain is currently under investigation. PMID- 12906738 TI - Interaction between anesthesia, gender, and functional outcome task following diffuse traumatic brain injury in rats. AB - A number of experimental and clinical studies have demonstrated that functional outcome following traumatic brain injury differs between males and females. Some studies report that females have a better outcome than males following trauma while others report the opposite. In experimental studies, some of the contradictory results may be due to the different experimental conditions, including type of anesthesia and the outcome measures employed. In the present study we have used three different anesthetic protocols and four different outcome measures to determine how these parameters interact and affect functional outcome following traumatic brain injury in male and female rats. Diffuse traumatic brain injury was induced in adult male and female animals using the impact-acceleration brain injury model. Mortality in female animals was no different than males when using halothane anesthesia, slightly better than males when using isoflurane anesthesia, but significantly worse than males under pentobarbital anesthesia. Female animals always performed better than males on rotarod tests of motor outcome, with this effect being unrelated to anesthetic effects. Conversely, in cognitive tests using the Barnes Maze, only isoflurane anesthetized females performed better than their male counterparts. Similarly, in an open field activity task, females always performed better than males after trauma, with isoflurane-anesthetized females also performing significantly better than the halothane-anesthetized female group after injury. Our results suggest that female animals do better than males after diffuse traumatic brain injury, although this observation is dependent upon the type of anesthesia and the functional task employed. Isoflurane is particularly protective in females, pentobarbital is deleterious to female outcome, while halothane anesthesia has the least influence on gender-related outcome. PMID- 12906739 TI - Moderate hypothermia improves neurobehavioral deficits after an epidural focal mass lesion in rodents. AB - The objective of this study was to evaluate the effects of a moderate, intraischemic hypothermia on the behavorial deficits up to 4 weeks after induction of a focal mass lesion. A focal epidural mass lesion was induced by an epidural balloon. The severity of the trauma was defined by the balloon volume and flattening of electroencephalography. Hypothermia (32 degrees C) was induced as soon as maximum balloon infIation was reached. Ischemia was extended over 30 min. After reperfusion, normothermic (n = 24) and hypothermic animals (n = 25) were monitored for 3 h followed by a rewarming of the cooled animals. Results were compared to sham-operated animals (n = 10). Behavioral deficits were assessed by postural reflex (PR), open field (OF), beam balance (BB), beam walking (BW), and water maze tests (WMT). MRI follow-up and histology was evaluated. Sham-operated rats showed normal test results. Rats with normothermia showed worsening of test performance (PR, p < 0.05; OF, p < 0.05; BB, p < 0.05; BW, p < 0.05; WMT, p < 0.05) compared to controls over the whole observation period. A significantly better behavioral outcome was observed in animals treated with hypothermia which showed no differences from controls 3-4 days after injury (PR, OF, BB, BW, WMT, p > 0.05). Lesion induced mortality was reduced in cooled animals but overall mortality rates were not influenced by this therapeutic measure. Neuronal cell loss in the CA1-CA4 region (p < 0.05) was reduced and the lesion size smaller (21%/p > 0.05) in hypothermic animals. Magnetic resonance imaging revealed that the lesion was more pronounced in the cortical grey matter after normothermia, whereas hypothermic animals showed more subcortical brain lacerations. In conclusion, intraischemic hypothermia significantly improved the behavioral outcome, and decreased lesion-induced mortality and the size of the lesion after an epidural focal mass lesion. PMID- 12906740 TI - Therapeutic vaccination for closed head injury. AB - Closed head injury often has a devastating outcome, partly because the insult, like other injuries to the central nervous system (CNS), triggers self destructive processes. During studies of the response to other CNS insults, it was unexpectedly discovered that the immune system, if well controlled, provides protection against self-destructive activities. Here we show that in mice with closed head injury, the immune system plays a key role in the spontaneous recovery. Strain-related differences were observed in the ability to harness a T cell-dependent protective mechanism against the effects of the injury. We further show that the trauma-induced deficit could be reduced, both functionally and anatomically, by post-traumatic vaccination with Cop-1, a synthetic copolymer used to treat patients with multiple sclerosis and found (using a different treatment protocol) to effectively counteract the loss of neurons caused by axonal injury or glutamate-induced toxicity. We suggest that a compound such as Cop-1 can be safely developed as a therapeutic vaccine to boost the body's immune repair mechanisms, thereby providing multifactorial protection against the consequences of brain trauma. PMID- 12906741 TI - Effects of cold injury-induced trauma in manganese superoxide dismutase-deficient mice. AB - Manganese superoxide dismutase (Mn-SOD, SOD2) is an inducible antioxidant localized to the mitochondria, which have been shown to be both the sites of superoxide anion (O(2)*-)) production and the target of free radical attacks. Knock-out mice with targeted disruption of Sod2 (SOD2-KO) are more susceptible to ischemic damage than their wild-type (WT) counterparts, showing increased loss of mitochondrial cytochrome c after trauma, but less apoptotic cell death in the first 24 h following controlled cortical injury. In this study, we sought to investigate whether oxidative stress plays a significant role in the development of secondary brain damage following cold injury-induced brain trauma (CIBT), a model of vasogenic edema. We first measured the levels of O(2)(*-) production 2 h after CIBT by means of in situ hydroethidine oxidation. We then examined lesion size, brain swelling, apoptosis by morphology and TUNEL-staining, neutrophil infiltration, and hemorrhage rates in both SOD2-KO and WT mice at 1, 3, and 7 days post-CIBT. We found no significant differences between SOD2-KO and WT littermates in any of the paradigms or endpoints studied. There was, however, a significant increase in hemorrhagic transformations in all animals that paralleled a robust inflammatory response at 3 days post insult compared with the 24-h endpoint. In the CIBT model used in this study, a 50% reduction in SOD2 activity did not appear to alter the injury response, suggesting that accumulation of free radicals does not play a significant role in secondary brain damage as previously thought with this particular model. PMID- 12906742 TI - Quercetin promotes functional recovery following acute spinal cord injury. AB - We tested the hypothesis that quercetin, a potent Fe(2+)-chelating flavonoid, would decrease secondary damage following spinal cord trauma. MRI studies using the relaxation of the T1 proton signal caused by Fe(2+) ions and the dose dependent reversal of this effect by addition of quercetin in aqueous solution were used to guide us to the dosage of quercetin to be used in animal experimentations. Forty-four male Wistar rats were used in two experimental series to test the hypothesis that administration of quercetin improves recovery of motor function after acute traumatic spinal cord injury. Animals were subjected to laminectomy and subjected to an extradural 40-g force clip compression for 5 sec at T7. Quercetin or saline was administered intraperitoneally 1 h after injury and then every 12 hr thereafter. Recovery of motor function was assessed using BBB scores at weekly intervals for 4 weeks. A dose of 2.5 micromoles quercetin/kg body weight did not result in significantly better functional outcome, whereas doses ranging from 5 to 100 micromoles quercetin/kg body weight resulted in a significantly better functional outcome with half or more of the animals walking, although with deficit; in contrast, no animals walked in the group of saline-treated animals. No significant differences in behavioral outcome were seen amongst the doses ranging from 5 to 100 micromol/kg, nor was there a difference if animals were treated for 4 or 10 days. Therapeutic outcome was coincident with more efficient iron clearance, suggesting that one possible mechanism whereby quercetin decreases secondary damage is through iron chelation. PMID- 12906743 TI - Neuronal sensitization and its behavioral correlates in a rat model of neuropathy are prevented by a cyclic analog of orphenadrine. AB - N-methyl-D-aspartic acid (NMDA) is an agonist at the homonymous receptor implicated in the development of neuronal sensitization and its behavioral correlates. An effective modulation of the NMDA effects, achieved also by uncompetitive antagonists, could contribute to controlling pain symptoms in several neuropathic syndromes. Because nefopam is a known analgesic derivative of orphenadrine and of its congener diphenhydramine, both uncompetitive NMDA receptor antagonists, we tested the effect of nefopam on the developing pain and neuronal anomalies in an animal model of chronic pain with NMDA receptor involvement. A single intraperitoneal injection of nefopam was administered twenty minutes prior to the chronic constriction injury of the sciatic nerve (CCI rats). In the first 10 days, nefopam (30 mg/kg) significantly decreased behavioral signs of neuropathic pain and the stimulus-evoked electrophysiological anomalies in recordings at 14 days, with only slight manifestation afterwards. The dose of 20 mg/kg was ineffective. Nefopam injected after constriction was ineffective. In normal non-operated rats, Nefopam had no effect on the electrophysiological and behavioral parameters. Iontophoretic nefopam (1 mM, 50 80 nA, positive current) in normal rats did not change the spontaneous neuronal activity, but reduced the mean response to noxious stimuli and the concurrent iontophoretic NMDA evoked activity. In CCI rats, iontophoretic nefopam did not significantly modify the spontaneous hyperactivity but reduced significantly both the frequency of the responses to noxious stimuli, and the duration of the afterdischarge. We propose that nefopam exerts a preventive analgesic effect, with a possible role in modulating NMDA receptor-mediated effects in central sensitization. PMID- 12906744 TI - GDNF and BDNF alter the expression of neuronal NOS, c-Jun, and p75 and prevent motoneuron death following spinal root avulsion in adult rats. AB - In the present study, we examined the effects of glial cell line-derived neurotrophic factor (GDNF), brain-derived neurotrophic factor (BDNF), ciliary neurotrophic factor (CNTF), and insulin growth factor (IGF-1) on adult motoneuron survival following spinal root avulsion. The expression of neuronal nitric oxide synthase (nNOS), c-Jun, and the low-affinity neurotrophin receptor (P75) following treatment with these neurotrophic factors was also examined. In control animals, approximately 80% of spinal motoneurons were nNOS positive at 3 weeks following the lesion, whereas in GDNF or BDNF treated animals no nNOS positive motoneurons were found at the same time point. Following injury and treatment with GDNF and BDNF increased numbers of motoneurons were c-Jun and P75 positive. By 6 weeks following the lesion, only approximately 28% of motoneurons persisted in control animals whereas about 90% of motoneurons survived injury following treatment with either GDNF or BDNF. In contrast, CNTF and IGF-1 were ineffective in either inhibiting nNOS expression or preventing motoneuron death. Our results provide in vivo evidence that the survival of injured adult mammalian motoneurons can be promoted by specific neurotrophic factors, and that this effect is associated with inhibition of nNOS expression and up-regulation of c-Jun and P75 expression. PMID- 12906745 TI - Rofecoxib: an update on physicochemical, pharmaceutical, pharmacodynamic and pharmacokinetic aspects. AB - Rofecoxib (MK-966) is a new generation non-steroidal anti-inflammatory agent (NSAID) that exhibits promising anti-inflammatory, analgesic and antipyretic activity. It selectively inhibits cyclooxygenase (COX)-2 isoenzyme in a dose dependent manner in man. No significant inhibition of COX-1 is observed with rofecoxib up to doses of 1000 mg. The pharmacokinetics of rofecoxib has been found to be complex and variable. Mean oral bioavailability after single dose of rofecoxib (12.5, 25 or 50 mg) is 93% with t(max) varying widely between 2 and 9 h. It is highly plasma-protein bound and is metabolized primarily by cytosolic reductases to inactive metabolites. Rofecoxib is eliminated predominantly by hepatic metabolism with a terminal half-life of approximately 17 h during steady state. Various experimental models and clinical studies have demonstrated rofecoxib to be superior, or at least equivalent, in anti-inflammatory, analgesic and antipyretic efficacy to comparator nonselective NSAIDs in osteoarthritis, rheumatoid arthritis and other pain models. Emerging evidence suggests that rofecoxib may also find potential use as supportive therapy in various pathophysiologic conditions like Alzheimer's disease, and in various malignant tumours and polyps, where COX-2 is overly expressed. Rofecoxib is generally well tolerated. Analysis of data pooled from several trials suggests that rofecoxib is associated with fewer incidences of clinically symptomatic gastrointestinal ulcers and ulcer complications vis-a-vis conventional NSAIDs. However, this gastropreserving effect may be negated by concurrent use of low-dose aspirin for cardiovascular risk reduction. Rofecoxib tends to show similar tolerability for renal and cardiothrombotic events as compared with nonnaproxen nonselective NSAIDs. No clinically significant drug interaction has been reported for rofecoxib except with diuretics, where it reverses their salt-wasting effect and thus can be clinically exploited in electrolyte-wasting disorders. There is only modest information about the physicochemical and pharmaceutical aspects of rofecoxib. Being poorly water soluble, its drug delivery has been improved using varied formulation approaches. Although it is stable in solid state, rofecoxib is photosensitive and base-sensitive in solution form with its degradation mechanistics elucidated. Analytical determinations of rofecoxib and its metabolites in biological fluids employing HPLC with varied types of detectors have been reported. Isolated studies have also been published on the chromatographic and spectrophotometric assay of rofecoxib and its degradants in bulk samples and pharmaceutical dosage forms. The current article provides an updated overview on the physicochemical, pharmaceutical, pharmacokinetic and pharmacodynamic vistas of rofecoxib. PMID- 12906746 TI - Effect of surfactant on fabrication and characterization of paclitaxel-loaded polybutylcyanoacrylate nanoparticulate delivery systems. AB - The feasibility of applying biodegradable polybutylcyanoacrylate (PBCA) nanoparticulate delivery systems (NDSs) for the controlled release of paclitaxel was investigated. Paclitaxel-loaded and unloaded PBCA-NDSs containing various surfactants (dextran 70, cholesterol, polyvinyl alcohol and lecithin) were prepared by anionic polymerization. The effects of surfactant (1% w/v), surfactant combination (1% w/v each), and surfactant concentration (0.05, 1.0 and 2.5% w/v) on PBCA-NDSs were evaluated and characterized by particle size, zeta potential, entrapment efficiency, and in-vitro paclitaxel release kinetics. The physicochemical characteristics of PBCA-NDSs incorporated with various surfactants were significantly improved compared with PBCA-NDS without any surfactant, by decreasing particle size at least 3-fold as well as by increasing the zeta potential up to 18-fold to minimize the agglomeration of nanoparticles. Moreover, PBCA-NDSs incorporated with various surfactants demonstrated higher entrapment efficiency of paclitaxel. Results from the in-vitro release kinetic studies indicated that a more controlled biphasic zero-order release pattern of paclitaxel was observed for PBCA-NDSs incorporated with various surfactants. Compared with dextran 70 and polyvinyl alcohol, the naturally occurring lipids, lecithin and cholesterol, indicated greater advantages in improving the physicochemical properties of PBCA-NDSs, in terms of smaller particle size, higher zeta potential and better drug entrapment efficiency, and better controlled release of paclitaxel, in terms of lower release rate and prolonged action from PBCA-NDSs. PMID- 12906747 TI - Formulation and evaluation of metronidazole acid gel for vaginal contraception. AB - In this study, the efficacy of metronidazole as a local spermicidal agent was investigated. The drug was formulated in a concentration of 5% as an acid gel for vaginal application as a local contraception. The minimum spermicidal concentration of metronidazole was evaluated in-vitro and found to be 0.2% w/v. The formulated gel showed instant immobilization and death of all sperm within 30 s. A clinical study was conducted to determine the drug concentrations in vaginal secretions of healthy women and women with symptoms suggesting genital tract infection every 15 min for a 1-h period after application of 1 g of the gel in the vagina. Drug concentrations in the infected group were significantly lower (P < 0.05) than those in the healthy group at all time intervals. The drug concentrations in vaginal secretions after 1 h of vaginal medication remained above the determined minimum spermicidal concentration (0.2%) in the two groups. Measurement of vaginal pH before and after medication revealed a significant drop (P < 0.05) in pH to its normal value in both healthy and infected groups as a result of gel application. In conclusion, the designed gel has potential advantages of achieving a long retention time and effective drug concentrations in the vagina for at least 1 h after application, and of maintaining vaginal pH within its normal range. PMID- 12906748 TI - Transdermal delivery of paracetamol for paediatric use: effects of vehicle formulations on the percutaneous penetration. AB - Paracetamol is a safe and effective analgesic and antipyretic agent, and is one of the most widely used medications for infants and children. The formulations currently available have been designed for oral and rectal administration. However, they are not practical in young patients with vomiting and diarrhoea, or in those who refuse to take the full dose. An alternative route of administration would be a significant contribution to the paediatric pharmacopoeia. The aim of this study was to develop a new transdermal system for optional therapeutic administration of paracetamol in infants and children. In-vivo studies were carried out in animals using a transdermal system of high-loaded, soluble paracetamol in a hydrogel patch, which was also tested in-vitro for 8 h. Although the beneficial contribution of glyceryl oleate to the transdermal penetration of paracetamol seemed to be significant in-vitro, it was shown to be insufficient in vivo. To improve the penetration of the drug, 4% PEG-40 stearate and 10% ethanol were incorporated as absorption enhancers into the dermal patches. A few hours after application of the improved patches to rats, plasma drug concentrations were elevated to levels comparable with those obtained after oral and subcutaneous administration of a high dose of paracetamol. Since plasma drug concentrations did not reach a constant steady state (as a peak or plateau) during the short-term animal experiments, longer pharmacokinetic studies in conscious animals are necessary. PMID- 12906749 TI - Spray-dried carbamazepine-loaded chitosan and HPMC microspheres: preparation and characterisation. AB - In this study, the potential of the spray-drying technique for preparing microspheres able to modify the release profile of carbamazepine was investigated. Low-, medium- and high-molecular-weight chitosan and hydroxypropyl methylcellulose (HPMC) in different drug-polymer ratios were used for the preparation of microspheres. The microspheres, characterized by X-ray powder diffractometry (XRD) and differential scanning calorimetry (DSC), were also studied with respect to particle size distribution, drug content and drug release. The results indicated that the entrapment efficiency (EE), as well as carbamazepine release profile, depended on polymeric composition and drug-polymer ratios of the microspheres prepared. The best entrapment efficiencies were obtained when chitosan of low-molecular-weight (CL) or HPMC were used for the microencapsulation. For all types of polymer used, the microspheres with low carbamazepine loading (6.3% w/w) showed better control of drug release than the microspheres with higher drug loadings. The HPMC microspheres showed the slowest carbamazepine release profile with no initial burst effect. Carbamazepine release profiles from ternary systems, carbamazepine-CL-HPMC microspheres, depended mostly on HPMC content and showed similar carbamazepine release profile as CL microspheres when HPMC content was low (9:1 CL-HPMC ratio, w/w). Otherwise, the carbamazepine release from CL-HPMC microspheres was remarkably faster than from either chitosan or HPMC microspheres. The release profile of carbamazepine from the microspheres was highly correlated with the crystalline changes occurring in the matrix. PMID- 12906750 TI - Controlled release of anti-cocaine catalytic antibody from biodegradable polymer microspheres. AB - Recent reports have shown that anti-cocaine catalytic monoclonal antibody 15A10 reduces the toxic effect of cocaine by increasing its breakdown to systemically inert products ecgonine methylester and benzoic acid. This study reports the microencapsulation of antibody 15A10 using biodegradable poly (lactic-glycolic) acid (PLGA) by double emulsion technique. Formulation parameters such as protein loading, polymer molecular weight and the presence of zinc carbonate were studied for their effects on in-vitro release of antibody from microspheres. The initial burst release was decreased by the reduction of the protein (as % of total ingredients) in the formulation. Although changing the polymer molecular weight did not cause a reduction in initial burst release, it was effective in improving the release rate. The inclusion of zinc carbonate in microsphere preparation resulted in increase in initial burst release. An in-vivo study in mice revealed the presence of antibody in blood up to ten days following subcutaneous injections. These data demonstrate a potential for a sustained-release formulation of monoclonal antibody 15A10 for treatment of cocaine addiction. PMID- 12906751 TI - Prediction of human pharmacokinetics from animal data and molecular structural parameters using multivariate regression analysis: volume of distribution at steady state. AB - The aim of this study was to develop a regression equation for predicting volume of distribution at steady state (Vd(ss)) in humans to enable application to various types of drugs using animal experimental data for rats and dogs and some molecular structural parameters. The Vd(ss) data for rats, dogs and humans of 64 drugs were obtained from literature. The compounds have various structures, pharmacological activities and pharmacokinetic characteristics. In addition, the molecular weight, calculated partition coefficient (clogP), and the number of hydrogen bond acceptors were used as possible descriptors related to the Vd(ss) in humans. Multivariate regression analyses, multiple linear regression analysis and the partial least squares (PLS) method were used to predict Vd(ss) in humans. Interaction terms were also introduced into the regression analysis to evaluate the non-linear relationship. For the data set used in the present study, PLS with quadratic term descriptors gave the best predictive performance. The PLS model using Vd(ss) data for only two animal species and using easily calculated structural parameters could generally predict Vd(ss) in humans better than an allometric method. In addition, the PLS model with only animal data gave almost the same predictive performance as the PLS model with quadratic term descriptors. This model may be easier to use and be practical in a realistic situation, and could predict Vd(ss) in humans better than the allometric method. PMID- 12906752 TI - A new animal model for evaluation of long-term growth rate over one month by rhGH/PLGA microcapsule formulations. AB - A new animal model to evaluate the long-term growth rate produced by a sustained release formulation of recombinant human growth hormone (rhGH) over one month was developed and the usefulness of our microcapsule formulations was demonstrated in this model. Long-term pharmacological effects by subcutaneous injection of microcapsules for sustained release of rhGH were evaluated in hypophysectomized (Hpx) rats treated with immunosuppressive agent along with hormone supplement. Copoly(DL-lactic/glycolic)acid (PLGA) microcapsules for sustained release of rhGH, a two-week sustained-release formulation (rhGH-SR-2W) and a one-month sustained-release formulation (rhGH-SR-1M), were prepared by a solid-in-oil-in water emulsion solvent evaporation technique. Body-weight gain, body-length gain and serum levels of rat insulin-like growth factor-I (rIGF-I) induced by subcutaneous injection of rhGH-SR were compared with those by daily injections of rhGH solution in Hpx rats for 35 days. Serum IGF-I levels in Hpx rats after the injection of rhGH-SR2W microcapsules were higher than those after daily injections of rhGH solution. Body-length gain, a new parameter, after single injection of rhGH-SR-1M microcapsules demonstrated the higher growth rate than that after daily injections of rhGH solution for 35 days. Thus, single injection of rhGH-SR microcapsules demonstrated long-term pharmacological effects greater than those by daily injections of rhGH solution in a newly developed model, immunosuppressed Hpx rats. PMID- 12906753 TI - Regional transport and metabolism of ropivacaine and its CYP3A4 metabolite PPX in human intestine. AB - The major aim of this study was to investigate the CYP3A4 metabolism and polarized transport of ropivacaine and its metabolite 2',6'-pipecoloxylidide (PPX) in tissue specimens from the human small and large intestine. Ropivacaine has been shown to be effective in the treatment of ulcerative colitis in human colon. This study was conducted using a modified Ussing-chamber technique with specimens from jejunum, ileum and colon collected from 11 patients. The local kinetics of ropivacaine and PPX were assessed from their concentration-time profiles in mucosal and serosal compartments. The permeability (P(app)) in the absorptive direction for both ropivacaine and PPX increased regionally in the order jejunum < ileum < colon. Ropivacaine was not found to be subjected to any carrier-mediated intestinal efflux. However, the CYP3A4 metabolite left the human enterocyte in a polarized manner and both the extent of CYP3A4 metabolism of ropivacaine and the extrusion of its metabolite to the mucosal chamber were more efficient in jejunum than in ileum. P-glycoprotein was probably not involved in the metabolite extrusion. No other metabolite than PPX was found. This in-vitro study with human intestinal tissues provides new mechanistic insights into regional transport and metabolism of drugs. PMID- 12906754 TI - Comparison of the growth-inhibitory effect of Hypericum perforatum L. extracts, differing in the concentration of phloroglucinols and flavonoids, on leukaemia cells. AB - In this study we compared, simultaneously, the growth-inhibitory effect of Hypericum perforatum L. extracts, containing various amounts of hyperforin (A 3.25%; B 2.21%; C 0.21% w/w) and flavonoids (A and B 5.3%; C 10% w/w), but closely same amounts of naphthodiantrones (0.3%) on two leukaemic cell lines K562 and U937 in the WST-1 assay. The GI50 (concentration of extracts which caused 50% of cell growth inhibition) for H. perforatum extracts analysed and characterized by HPLC for their biologically active constituents was 248.3-621.3 microg mL(-1) in K562 and 378.2-911.7 microg mL(-1) in U937 cells. The corresponding values of the three main groups were 1.6-3.9 microM naphthodianthrones, 1.0-40.7 microM phloroglucinols and 30.5-68.5 microM flavonoids. The results of this study supported the hypothesis that, apart from hyperforin and flavonoids, other components of the extract could be involved in its growth-inhibitory effect that it exerts without light activation. PMID- 12906755 TI - Screening pharmaceutical preparations containing extracts of turmeric rhizome, artichoke leaf, devil's claw root and garlic or salmon oil for antioxidant capacity. AB - Pharmaceutical preparations derived from natural sources such as vegetables often contain compounds that contribute to the antioxidant defence system and apparently play a role in the protection against degenerative diseases. In the present study, commercial preparations containing extracts of turmeric, artichoke, devil's claw and garlic or salmon oil were investigated. The products were divided into fractions of different polarity, and their antioxidant activity was determined using the Trolox equivalent antioxidant capacity (TEAC) assay. This test is based on the efficacy of the test material to scavenge 2,2'-azino bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) derived radicals. Total phenols were determined in all fractions as well as specific carotenoids in the most lipophilic fraction to assess their contribution to the antioxidant activity. For comparison, the radical scavenging effect of selected constituents of the extracts such as curcumin, luteolin, kaempferol, chlorogenic acid, harpagoside, beta-carotene and alpha-tocopherol was investigated and compared with that of Trolox. Curcumin, luteolin, kaempferol, chlorogenic acid and beta-carotene showed an antioxidant activity superior to Trolox in the TEAC assay; harpagoside was barely active. All fractions of the turmeric extract preparation exhibited pronounced antioxidant activity, which was assigned to the presence of curcumin and other polyphenols. The antioxidant activity corresponding to the artichoke leaf extract was higher in the aqueous fractions than in the lipophilic fractions. Similarly, devil's claw extract was particularly rich in water-soluble antioxidants. Harpagoside, a major compound in devil's claw, did not contribute significantly to its antioxidant activity. The antioxidant capacity of the garlic preparation was poor in the TEAC assay. That of salmon oil was mainly attributed to vitamin E, which is added to the product for stabilization. In all test preparations, the antioxidant activity was significantly correlated with the content of total phenolic compounds. PMID- 12906756 TI - Synergistic Capsicum-tea mixtures with anticancer activity. AB - We have demonstrated a synergy between a decaffeinated green tea concentrate and a vanilloid-containing Capsicum preparation obtained commercially. At a ratio of 25 parts green tea concentrate to 1 part Capsicum preparation, the resultant product exhibited efficacy in the killing of cancer cells in culture 100-times that of green tea on a weight basis. These studies were guided by assays of the putative catechin-vanilloid target protein tNOX, a cell surface growth-related enzymatic activity specific to cancer. The activity of the protein target was inhibited by the tea catechins and the Capsicum vanilloids. As with growth, the tea and Capsicum preparations evaluated were synergistic in their inhibition of the target enzymatic activity. PMID- 12906757 TI - Inhibitory effect of erythromycin on potassium currents in rat ventricular myocytes in comparison with disopyramide. AB - Disopyramide, a class Ia antiarrhythmic agent, has been reported to induce torsades de pointes (TdP) associated with excessive QT prolongation in electrocardiogram (ECG), especially when concomitantly administered with erythromycin, a macrolide antibiotic agent. In this study, we have evaluated the effects of erythromycin on action potential duration (APD) and potassium currents in rat ventricular myocytes in comparison with disopyramide. We have evaluated the relationship between in-vitro potassium current inhibition and in-vivo QT prolongation observed in a previous study. Action potentials and membrane potassium currents, including delayed rectifier current (I(K)) and transient outward current (I(to)), were recorded using a whole-cell patch clamp method in enzymatically-dissociated ventricular cells. Erythromycin and disopyramide prolonged APD in a concentration-dependent manner. Disopyramide (10-100 microM) and erythromycin (100 microM) led to increases in the APD at 90% repolarization level. Disopyramide reduced I(K) (IC50 = 37.2 +/- 0.17 microM) and I(to) (IC50 = 20.9 +/- 0.13 microM) while erythromycin reduced I(K) (IC50 = 60.1 +/- 0.29 microM) but not I(to). The observed prolongation of APD might be ascribed to the inhibition of potassium currents. Erythromycin produced the prolongation of APD and the inhibition of potassium currents with a lag time after addition of the drugs, which suggested that erythromycin might not reach potassium channels from outside the ventricular cells. The potency of disopyramide was almost equivalent under in-vitro and in-vivo conditions. However, potency of erythromycin in-vitro was far weaker than that in-vivo reported in a previous study, presumably due to a difference in the uptake of erythromycin into ventricular myocytes between in vivo and in-vitro conditions. Therefore, when drug-induced risks of QT prolongation are to be evaluated, the difference of potencies between in-vitro and in-vivo should be taken into consideration. PMID- 12906758 TI - Stereospecific reduction of the original anticancer drug oracin in rat extrahepatic tissues. AB - The liver is the major site of drug metabolism in the body. However, many drugs undergo metabolism in extrahepatic sites and in the gut wall and lumen. In this study, the distribution and activity of reductases in rat that reduced potential cytostatic oracin to its principal metabolite 11-dihydrooracin (DHO) were investigated. The extension and stereospecificity of oracin reduction to DHO were tested in microsomal and cytosolic fractions from the liver, kidney, heart, lung and wall of small intestine, caecum and large intestine. Intestinal bacterial reduction of oracin was studied as well. The amount of DHO enantiomers was measured by HPLC with Chiralcel OD-R as chiral column. Reductive biotransformation of oracin was mostly stereospecific for (+)-DHO, but the enantiomeric ratio differed significantly among individual tissues and subcellular fractions (from 56% (+)-DHO in heart microsomes to 92% (+)-DHO in liver cytosol). Stereospecificity for (-)-DHO (60%) was observed in bacterial oracin reduction in the lumen of small intestine, caecum and large intestine. Shift of the (+)-DHO/(-)-DHO enantiomeric ratio from 90:10 (in liver subcellular fractions) to 60:40 (in-vivo) clearly demonstrated the importance of the contribution of extrahepatic metabolism to the total biotransformation of oracin to DHO. PMID- 12906759 TI - Contribution of organic anion transporting polypeptide OATP-C to hepatic elimination of the opioid pentapeptide analogue [D-Ala2, D-Leu5]-enkephalin. AB - The objective of this study was to examine the transport activity of the human organic anion transporter OATP-C (SLC21A6) for oligopeptides that are eliminated rapidly from the systemic circulation. We focused on an opioid peptide analogue, [D-Ala(2), D-Leu(5)]-enkephalin (DADLE), a linear pentapeptide modified to be stable. [(3)H]DADLE was taken up by rat isolated hepatocytes in a saturable manner and highly accumulated in the liver after intravenous administration to rats. The uptake of [(3)H]DADLE by the isolated hepatocytes was inhibited by several organic anions and pentapeptides, but not by tetra- or tripeptides. When OATP-C was expressed in Xenopus laevis oocytes, a significant increase in uptake of [(3)H]DADLE was observed. Moreover, the inhibitory effects of various compounds, including some peptides, on [(3)H]estrone-3-sulfate uptake by OATP-C were similar to those observed in [(3)H]DADLE uptake by rat isolated hepatocytes. In conclusion, it was demonstrated that OATP-C contributes to the rapid hepatic excretion of peptides and peptide-mimetic drugs. PMID- 12906760 TI - Hypochlorite scavenging activity of hydroxycinnamic acids evaluated by a rapid microplate method based on the measurement of chloramines. AB - Scavengers of hypochlorite (XOCl) could have beneficial effects in diseases in which this oxidant plays a pathogenic role. It has been reported that ferulic acid and chlorogenic acid, the quinic ester of caffeic acid, are good hypochlorite scavengers, but a systematic evaluation of the naturally occurring hydroxycinnamic acids (HCAs), which these substances belong to, has not been performed yet. Thus, in this work we studied, by two different in-vitro methods, the antioxidant activity of five HCAs: p-coumaric acid, ferulic acid, sinapinic acid, caffeic acid and chlorogenic acid. The methods applied in this study were based on the oxidation of human serum albumin (HSA) by XOCl, a new microplate method based on the measurement of chloramines and a previously described carbonyl assay. Firstly, lysine-derived chloramines, in the presence or absence of the HCAs, were detected using 5-thio-2-nitrobenzoic acid (TNB), measuring the absorbance at 415 nm by a microplate reader. To remove excess XOCl, Trolox, a known XOCl scavenger, was added before TNB. Secondly, lysine-derived carbonyls, in the presence or absence of the HCAs, were detected by using 2,4 dinitrophenylhydrazine. Hydroxycinnamic acids appeared active (caffeic >/= sinapinic > chlorogenic congruent with ferulic > p-coumaric acid) by both methods, suggesting possible pharmacological applications for these compounds, which are present at high concentrations in the plant kingdom. PMID- 12906761 TI - Interactions of a series of novel spiropyranocoumarin derivatives with reactive oxygen species. AB - A series of new spiro-substituted pyranocoumarin derivatives have been synthesized starting from the commercially available 7-hydroxycoumarin and the conformation of the pyran ring was investigated. The antioxidant activity of the compounds was evaluated in-vitro, by means of three different tests: the interaction with the stable free radical 1,1-diphenyl-2-picrylhydrazyl (DPPH), the competition with DMSO for hydroxyl radicals scavenging ability and the quenching of superoxide anions generated by the enzymic xanthine-xanthine oxidase system. In the DPPH test the spiroadamantane derivative 13 was the most active and possessed a 40% inhibition at a concentration of 400 microM. All compounds successfully compete with DMSO for hydroxyl radicals generated by the Fe(3+)/ascorbic acid system. Compound 13 inhibited the oxidation of DMSO (3.125 mM) by 93% at 2 mM and by 71% at 0.25 mM. The corresponding second-order rate constants have been estimated and all compounds demonstrated higher rate constants compared with the reference compounds, 7-hydroxycoumarin and mannitol. Derivatives possessing extended conjugation showed the highest inhibitory activity for superoxide anions generated by the xanthine-xanthine oxidase system, although the results of this experiment possessed partial parallelism with the results observed in the other two tests. The overall obtained data indicate that the size of the different spiro- substituents influence the degree of free radical scavenging and demonstrate the importance of extended conjugation for the antioxidant activity. Due to its multiple mechanism of protective action, derivative 13 may serve as a lead for the development of analogues that could be useful for the treatment of pathophysiological processes dependent upon reactive oxygen species. PMID- 12906763 TI - Neurotrophic factors protect myelin from attack. PMID- 12906764 TI - EAE in the common marmoset Callithrix jacchus. AB - The common marmoset Callithrix jacchus is a primate phylogenetically close to humans which, when immunized with myelin proteins, is susceptible to a form of experimental autoimmune encephalomyelitis (EAE) that resembles multiple sclerosis (MS). Neuropathological features of marmoset EAE, including inflammation, demyelination and axonal injury, are strikingly similar to findings in the human disease and are the final result of a joint autoimmune attack by myelin-specific T and B cells. The molecular and functional similarity of the marmoset immune system, together with the availability of diagnostic tools that can be used in humans (such as magnetic resonance imaging), makes the marmoset EAE a unique model with which to evaluate the safety and efficacy of therapeutic strategies for the treatment of MS. PMID- 12906765 TI - Pain and MS. AB - Pain is frequently evident in the course of multiple sclerosis (MS) and is estimated to occur in approximately 50% of patients. The incidence of pain has no apparent correlation to disease severity. Moreover, a comprehensive definition of pain has not been established, thus making the evaluation of this chronic, evolving symptom difficult. On the basis of its pathophysiology, pain can occur as a consequence of neurological impairment and disability, somatic pain or because of neurological damage. Although there are few randomized trials for pain in MS, new therapeutic strategies are now available and interest in the symptomatic treatment of MS is growing. PMID- 12906766 TI - Commentary on J Clausen's review--endogenous retroviruses in MS. PMID- 12906767 TI - Endogenous retroviruses and MS: using ERVs as disease markers. AB - Multiple sclerosis (MS) has an unknown cause, but its epidemiology suggests an interplay between environmental factors, possibly including viruses, and genetic components. Endogenous retroviruses (ERVs) are elements of the human genome that potentially may act as either genetic markers for polymorphisms related to MS, or markers of environmental/endogenous stress. Activation of the ERVs HERV-H/RGH, HERV-W and ERV-9 was reported when specific cell types (mainly B cells) from MS patients were cultivated in vitro. Viral RNA from these ERVs has been detected by reverse-transcriptase polymerase chain reaction in sera/plasma and brain tissues from MS patients, although not exclusively from these patients. ERVs play unknown roles in MS: their activation may represent an inflammatory cytokine-mediated epiphenomenon; alternatively, preliminary evidence suggests that specific ERVs may act as auto-, super- or neoantigens with the potential to enhance inflammatory responses or induce autoimmune reactions. ERVs that occur in few copies in the human genome (e.g. ERV-3 and human endogenous retrovirus, HRES-1) may show polymorphic patterns in MS. Studies show that the sequences encoding the envelope protein of ERV-3 are polymorphic to a degree where it becomes impossible to link them with MS. In contrast, the HRES-1 long terminal repeat sequence has a polymorphic pattern with haplotypes characteristic of MS. Haplotypes from non-MS control groups were identical between different topographic areas, but haplotypes from MS patients were different, depending on the population. PMID- 12906768 TI - Revisiting The pathogenesis of multiple sclerosis revisited. PMID- 12906769 TI - Controversies across continents. Report from the MS Forum Symposium, September 2002, Baltimore, USA. AB - The personal views of speakers from North America and Europe concerning the diagnosis and management of people with multiple sclerosis (MS) were presented at an MS Forum symposium before ACTRIM/ECTRIMS Meeting in Baltimore, USA. There were more areas of agreement than controversy and the overall impression was that MS is managed similarly across the continents. Four topics were discussed: diagnosing the clinically isolated syndrome; guidelines for using disease modifying therapy; people who fail to respond to disease-modifying treatment; and treating the patient with rapid deterioration to disability. PMID- 12906772 TI - Immune disorders caused by defects in the caspase cascade. AB - In the immune system, lymphocyte activation by antigen is followed by cell proliferation and induction of effector functions. Subsequently, physiologic cell death signals are induced, resulting in removal of expanded effector-cell populations, to maintain homeostasis. Caspases are intracellular participants in both activation responses and cell death by apoptosis. Targets of caspases include inflammatory activators and also other members of the caspase family that mediate apoptosis. Caspase-8 and caspase-10 participate in the protease cascade following cell surface CD95 engagement by its ligand. Humans with defects in these caspases were initially evaluated for the autoimmune lymphoproliferative syndrome because of their spleen and lymph node enlargement. Although both caspase-8- and caspase-10-deficient individuals had impaired apoptosis, those with caspase-8 deficiency, who also had immunodeficiency, had additional defects in activation of lymphocytes and natural killer cells. These disorders help to define the importance and specificity of the caspase proteases in intracellular signaling pathways. PMID- 12906773 TI - Molecular basis and therapy of disorders associated with chronic neutropenia. AB - There have been many recent advances in our understanding of the molecular basis of neutropenia disorders, primarily through advances in genetic analysis of inherited disorders. Molecular and cellular studies now suggest that accelerated apoptosis of neutrophil precursors in the bone marrow is the common pathophysiologic mechanism. Severe congenital neutropenia and cyclic neutropenia, both usually inherited as autosomal-dominant disorders, are caused by mutations in the neutrophil elastase gene. Myelokathexis is attributed to the downregulation of the bcl-x protein, but the genetic basis is not yet known. The genes for several diseases with more complex phenotypes (eg, glycogen storage disease type 1b, Chediak-Higashi syndrome, Shwachman-Diamond syndrome, dyskeratosis congenita, Griscelli syndrome, Barth syndrome, and Wiskott-Aldrich syndrome) have all been identified recently. The molecular mechanisms for most acquired disorders causing neutropenia (eg, idiopathic neutropenia, pure white cell aplasia, myelodysplasia, and aplastic anemia) are not yet known. Granulocyte colony stimulating factor (G-CSF) is effective treatment for several of these conditions. Through better understanding of these disorders, we anticipate that better treatments will be found in the future. PMID- 12906774 TI - Secondary immune deficiencies associated with biological therapeutics. AB - Biological therapeutics have been utilized in the past several years for the treatment of various autoimmune, cardiovascular, and neoplastic conditions. As these agents target both pathologic and physiologic components of inflammatory and immune systems, the scientific community has been challenged to identify potential secondary immune deficiencies associated with their use. This article will focus on biological agents used for the treatment of rheumatoid arthritis, including tumor necrosis factor-a inhibitors, cytotoxic T lymphocyte-associated antigen-4 (CTLA-4 Ig), rituximab, and interleukin-1 receptor agonist (IL-1Ra). Both clinical and in vitro studies have been done to explore the potential immunomodulatory and immunosuppressive effects of these agents. PMID- 12906775 TI - Immune deficiencies due to defects in cytokine signaling. AB - Severe combined immunodeficiency (SCID) represents a syndrome comprising the most severe forms of inherited immunodeficiencies. Defects in cytokine signaling pathways can result in impaired development of lymphoid cells and/or defective functioning of these cells, and most cases of SCID result from defective signaling through the common cytokine receptor g chain (g(c)) or associated molecules and signaling pathways. Studies of these patients and the analysis of gene-targeted mice provide insight into the underlying signaling defects in inherited immunodeficiencies. The identification of the genetic defects in humans with SCID provides the basis for future therapies for these patients. More subtle deficiencies in cytokine signaling have also been found as causes of other forms of immunodeficiency, and the knowledge learned could lead to novel approaches to antimicrobial therapy. PMID- 12906776 TI - Endogenous production of antimicrobial peptides in innate immunity and human disease. AB - Antimicrobial peptides are diverse and evolutionarily ancient molecules produced by all living organisms. Peptides belonging to the cathelicidin and defensin gene families exhibit an immune strategy as they defend against infection by inhibiting microbial survival, and modify hosts through triggering tissue specific defense and repair events. A variety of processes have evolved in microbes to evade the action of antimicrobial peptides, including the ability to degrade or inactivate antimicrobial peptides, or suppress host production of the peptide in response to infection. Animal models and clinical investigations have shown that an absence of cathelicidin or defensin antimicrobials can lead to disease. In this article, we review important recent advances in understanding the biology of antimicrobial peptides and their role in normal immunity and human disease. PMID- 12906780 TI - Fungal allergens. AB - Many fungi are capable of causing IgE-mediated hypersensitivity in humans. However, the most predominant fungi implicated in allergy belong to the genera Aspergillus, Alternaria, Cladosporium, and Penicillium. Pure and relevant allergens are essential for diagnosis as well as for understanding the immunopathogenesis of the disease. Until recently, pure and standardizable antigens from fungi were not available. In recent years, many recombinant allergens have been produced by molecular cloning. Using these allergens, novel methods are being developed to improve diagnosis of mold-induced allergy. By understanding the immunopathogenesis of allergens, new avenues might open up leading to improved patient care, including immunotherapy and vaccination. This review covers the current status of fungal allergens, their role in reliable immunodiagnosis, and their probable use in immunotherapy and vaccination. PMID- 12906781 TI - Environmental control for fungal allergen exposure. AB - With our limited knowledge of the mold allergens, we must rely on common sense to reduce environmental exposure for patients who are allergic to mold. We understand that the first step is moisture control. Appropriate building design, keeping rainwater and ground water away from the interior, accomplishes this. In addition, the heating ventilation and air conditioning system must be appropriately designed and maintained. Functional maintenance of inside water sources such as free water from plumbing, appliances, and showers can prevent damaging leaks. Indoor humidity or water vapor must be controlled and maintained to prevent condensation on walls or in microenvironments, such as attics, bedrooms, basements, and beneath wall-to-wall carpeting. Few abatement trials have been published, but several suggest that such measures can reduce mold exposure. PMID- 12906782 TI - Exposure, sensitization, and mechanisms of fungus-induced asthma. AB - Healthy individuals are continuously exposed to fungal biomass, which includes live and dead spores and fungal debris that is entrapped in the airways. In patients with asthma and/or atopy, exposure to fungal biomass might result in age dependent sensitization and asthmatic reactions. Interaction with Toll-like receptors (TLRs) of the innate immune defense (alveolar macrophages and epithelial cells) and protease-activated receptors (PARs) determine the effectiveness of elimination of fungal material. The association of sensitization to Alternaria with severe asthma is discussed in relation to the age-dependent sensitization, rate of release of allergens from spores, and activity of its proteases. A model is described concerning the influence of polymorphic genes for airway hyperresponsiveness (AHR) and atopy, showing a cumulating influence on susceptibility for allergen-induced asthma, and explaining that fungus-induced airway obstruction is mainly associated with more severe asthma. PMID- 12906783 TI - Toxic and other non-IgE-mediated effects of fungal exposures. AB - There are more than 100000 recognized species of fungi, comprising 25% of the biomass of the earth. Allergic, IgE-induced, manifestations of airborne fungi are common, whereas non-IgE manifestations are rare. Recently, much focus has been placed on the non-IgE-mediated effects of various molds, including hypersensitivity pneumonitis, infectious disease, and mycotoxicoses. Hypersensitivity pneumonitis is a clinical syndrome associated with systemic and interstitial lung disease that occurs in susceptible individuals following fungal inhalation. Most fungi are not pathogenic to man; however, certain fungi are capable of infecting immunocompetent individuals. Although mycotoxins and exposure to mycotoxins ("toxic mold syndrome") are implicated in causing numerous, nonspecific, systemic symptoms, currently, there is no scientific evidence to support the allegation that human health is affected by inhaled mycotoxins. However, if mold is discovered in a home, school, or office setting, the source should be investigated and appropriate remediation undertaken to minimize structural damage and potential allergic sensitization. PMID- 12906784 TI - Immunotherapy in fungal allergy. AB - For decades airborne fungal spores have been implicated as causative factors in respiratory allergy. Exposure to high atmospheric spore counts and sensitization to specific fungal allergens have been associated with severe asthma, mainly in young adults. Although the prevalence of sensitization to commercial fungal extracts is approximately 3% in epidemiologic studies, in selected patients, particularly with asthma, the sensitization rate might increase to 30%. Of the estimated number of more than 1 million of different fungal species, approximately 80 fungi have been connected with respiratory allergy. Currently, diagnosis and specific therapy of fungal allergy is hampered by the poor quality of most of the commercially available extracts. Clinical efficacy of specific immunotherapy with fungal extracts has been shown in 79 actively treated patients in four controlled trials, with only two fungal species, namely Alternaria alternata and Cladosporium herbarum. The use of recombinant fungal allergens might create new prospects in diagnosis and specific immunotherapy for fungal allergy. PMID- 12906785 TI - Tuberous sclerosis complex gene products, Tuberin and Hamartin, control mTOR signaling by acting as a GTPase-activating protein complex toward Rheb. AB - BACKGROUND: Tuberous Sclerosis Complex (TSC) is a genetic disorder that occurs through the loss of heterozygosity of either TSC1 or TSC2, which encode Hamartin or Tuberin, respectively. Tuberin and Hamartin form a tumor suppressor heterodimer that inhibits the mammalian target of rapamycin (mTOR) nutrient signaling input, but how this occurs is unclear. RESULTS: We show that the small G protein Rheb (Ras homolog enriched in brain) is a molecular target of TSC1/TSC2 that regulates mTOR signaling. Overexpression of Rheb activates 40S ribosomal protein S6 kinase 1 (S6K1) but not p90 ribosomal S6 kinase 1 (RSK1) or Akt. Furthermore, Rheb induces phosphorylation of eukaryotic initiation factor 4E binding protein 1 (4E-BP1) and causes 4E-BP1 to dissociate from eIF4E. This dissociation is completely sensitive to rapamycin (an mTOR inhibitor) but not wortmannin (a phosphoinositide 3-kinase [PI3K] inhibitor). Rheb also activates S6K1 during amino acid insufficiency via a rapamycin-sensitive mechanism, suggesting that Rheb participates in nutrient signaling through mTOR. Moreover, Rheb does not activate a S6K1 mutant that is unresponsive to mTOR-mediated signals, confirming that Rheb functions upstream of mTOR. Overexpression of the Tuberin-Hamartin heterodimer inhibits Rheb-mediated S6K1 activation, suggesting that Tuberin functions as a Rheb GTPase activating protein (GAP). Supporting this notion, TSC patient-derived Tuberin GAP domain mutants were unable to inactivate Rheb in vivo. Moreover, in vitro studies reveal that Tuberin, when associated with Hamartin, acts as a Rheb GTPase-activating protein. Finally, we show that membrane localization of Rheb is important for its biological activity because a farnesylation-defective mutant of Rheb stimulated S6K1 activation less efficiently. CONCLUSIONS: We show that Rheb acts as a novel mediator of the nutrient signaling input to mTOR and is the molecular target of TSC1 and TSC2 within mammalian cells. PMID- 12906786 TI - VA opsin, melanopsin, and an inherent light response within retinal interneurons. AB - BACKGROUND: Although photoreception is best understood in rods and cones, it is increasingly clear that these are not the only photoreceptive cells of the vertebrate retina. While considerable attention has been paid to the role of melanopsin in the generation of intrinsic light sensitivity in the retinal ganglion cells of mammals, nothing is known about the photoreceptive capacity of the horizontal cells of the fish retina in which both VA opsin and melanopsin are expressed. As yet, there has been little more than speculation as to the physiological function of these opsins within local retinal circuit neurons. RESULTS: VA opsin and melanopsin have been isolated and localized within the well characterized cyprinid retina of the roach (Rutilus rutilus). Parallel electrophysiological studies identified a novel subtype of horizontal cell (HC RSD) characterized by a depolarizing response that fits an opsin photopigment with a lambda(max) of 477 nm. The HC-RSD cells mediate responses to light that are characterized by long integration times, well beyond those observed for rods and cones. Significantly, HC-RSD responses persist when the conventional photoreceptor inputs are saturated by background light. CONCLUSIONS: The syncytium of coupled horizontal cells has long been considered to provide a signal of overall retinal irradiance. Our data suggest that this light information is, at least in part, derived from a population of intrinsically photosensitive VA opsin and/or melanopsin horizontal cells. PMID- 12906787 TI - Phosphoinositide 3-kinase is required for process outgrowth and cell polarization of gastrulating mesendodermal cells. AB - BACKGROUND: During vertebrate gastrulation, cell polarization and migration are core components in the cellular rearrangements that lead to the formation of the three germ layers, ectoderm, mesoderm, and endoderm. Previous studies have implicated the Wnt/planar cell polarity (PCP) signaling pathway in controlling cell morphology and movement during gastrulation. However, cell polarization and directed cell migration are reduced but not completely abolished in the absence of Wnt/PCP signals; this observation indicates that other signaling pathways must be involved. RESULTS: We show that Phosphoinositide 3-Kinases (PI3Ks) are required at the onset of zebrafish gastrulation in mesendodermal cells for process formation and cell polarization. Platelet Derived Growth Factor (PDGF) functions upstream of PI3K, while Protein Kinase B (PKB), a downstream effector of PI3K activity, localizes to the leading edge of migrating mesendodermal cells. In the absence of PI3K activity, PKB localization and cell polarization are strongly reduced in mesendodermal cells and are followed by slower but still highly coordinated and directed movements of these cells. CONCLUSIONS: We have identified a novel role of a signaling pathway comprised of PDGF, PI3K, and PKB in the control of morphogenetic cell movements during gastrulation. Furthermore, our findings provide insight into the relationship between cell polarization and directed cell migration at the onset of zebrafish gastrulation. PMID- 12906788 TI - Calcium imaging reveals a network of intrinsically light-sensitive inner-retinal neurons. AB - BACKGROUND: Mice lacking rod and cone photoreceptors (rd/rd cl) are still able to regulate a range of responses to light, including circadian photoentrainment, the pupillary light reflex, and suppression of pineal melatonin by light. These data are consistent with the presence of a novel inner-retinal photoreceptor mediating non-image-forming irradiance detection. RESULTS: We have examined the nature and extent of intrinsic light sensitivity in rd/rd cl retinae by monitoring the effect of light stimulation (470 nm) on intracellular Ca(2+) via FURA-2 imaging. Using this approach, which does not rely on pharmacological or surgical isolation of ganglion cells from the rod and cone photoreceptors, we identified a population of light-sensitive neurons in the ganglion cell layer (GCL). Retinal illumination induced an increase of intracellular Ca(2+) in approximately 2.7% of the neurons. The light-evoked Ca(2+) fluxes were dependent on the intensity and duration of the light stimulus. The light-responsive units formed an extensive network that could be uncoupled by application of the gap junction blocker carbenoxolone. Three types of light-evoked Ca(2+) influx were observed: sustained, transient, and repetitive, which are suggestive of distinct functional classes of GCL photoreceptors. CONCLUSIONS: Collectively, our data reveal a heterogeneous syncytium of intrinsically photosensitive neurons in the GCL coupled to a secondary population of light-driven cells, in the absence of rod and cone inputs. PMID- 12906789 TI - Elm1p is one of three upstream kinases for the Saccharomyces cerevisiae SNF1 complex. AB - BACKGROUND: The yeast SNF1 protein kinase and the mammalian AMP-activated protein kinase are highly conserved heterotrimeric complexes that are "metabolic master switches" involved in the switch from fermentative/anaerobic to oxidative metabolism. They are activated by cellular stresses that deplete cellular ATP, and SNF1 is essential in the response to glucose starvation. In both cases, activation requires phosphorylation at a conserved threonine residue within the activation loop of the kinase domain, but identifying the upstream kinase(s) responsible for this has been a challenging, unsolved problem. RESULTS: Using a library of strains that express 119 yeast protein kinases as GST fusions, we identified Elm1p as the sole kinase that could activate the kinase domain of AMP activated protein kinase in vitro. Elm1p also activated the purified SNF1 complex, and this correlated with phosphorylation of Thr210 in the activation loop. Removal of the C-terminal domain increased the Elm1p kinase activity, indicating that it is auto-inhibitory. Expression of activated, truncated Elm1p from its own promoter gave a constitutive pseudohyphal growth phenotype that was rescued by deletion of SNF1, showing that Snf1p was acting downstream of Elm1p. Deletion of ELM1 does not give an snf- phenotype. However, Elm1p is closely related to Pak1p and Tos3p, and a pak1Delta tos3Delta elm1Delta triple mutant had an snf1- phenotype, i.e., it would not grow on raffinose and did not display hyperphosphorylation of the SNF1 target, Mig1p, in response to glucose starvation. CONCLUSIONS: Elm1p, Pak1p, and Tos3p are upstream kinases for the SNF1 complex that have partially redundant functions. PMID- 12906790 TI - Light-mediated activation of Rac-1 in photoreceptor outer segments. AB - Small GTP binding proteins regulate diverse biological processes including gene expression, cytoskeleton reorganization, and protein and vesicular transport. While small GTPases have been investigated in a wide variety of cells, few studies have addressed their role in photoreceptors. In vertebrate retinal rods, the light stimulus is transmitted from rhodopsin via the pathway mediated by the heterotrimeric G protein transducin. To increase their sensitivity to light, photoreceptors accumulate remarkably high concentrations of rhodopsin and transducin in specialized cellular compartments, the outer segments (OS). Transport of these proteins from the inner segments is regulated by the small GTPases Rab6 and Rab8, which do not enter OS. Here, we asked if small G proteins have other functions in photoreceptors. We show that OS contain the small GTPase Rac-1, a member of the Rho family. In contrast to other cells, Rac-1 in OS is exclusively associated with the membranes and resides in lipid rafts. Most importantly, Rac-1 is activated by light. This activation is specifically blocked by a synthetic peptide corresponding to the Asn-Pro-X-X-Tyr motif found in rhodopsin, and Rac-1 coprecipitates with rhodopsin on Concanavalin A Sepharose. These data provide the first direct evidence for the existence of a novel pathway activated by rhodopsin. PMID- 12906791 TI - A genome-wide screen identifies 27 genes involved in transposon silencing in C. elegans. AB - Transposon jumps are a major cause of genome instability. In the C. elegans strain Bristol N2, transposons are active in somatic cells, but they are silenced in the germline, presumably to protect the germline from mutations. Interestingly, the transposon-silencing mechanism shares factors with the RNAi machinery. To better understand the mechanism of transposon silencing, we performed a genome-wide RNAi screen for genes that, when silenced, cause transposition of Tc1 in the C. elegans germline. We identified 27 such genes, among which are mut-16, a mutator that was previously found but not identified at the molecular level, ppw-2, a member of the argonaute family, and several factors that indicate a role for chromatin structure in the regulation of transposition. Some of the newly identified genes are also required for cosuppression and therefore represent the shared components of the two pathways. Since most of the newly identified genes have clear homologs in other species, and since transposons are found from protozoa to human, it seems likely that they also protect other genomes against transposon activity in the germline. PMID- 12906792 TI - Two neuronal, nuclear-localized RNA binding proteins involved in synaptic transmission. AB - While there is evidence that distinct protein isoforms resulting from alternative pre-mRNA splicing play critical roles in neuronal development and function, little is known about molecules regulating alternative splicing in the nervous system. Using Caenorhabditis elegans as a model for studying neuron/target communication, we report that unc-75 mutant animals display neuroanatomical and behavioral defects indicative of a role in modulating GABAergic and cholinergic neurotransmission but not neuronal development. We show that unc-75 encodes an RRM domain-containing RNA binding protein that is exclusively expressed in the nervous system and neurosecretory gland cells. UNC-75 protein, as well as a subset of related C. elegans RRM proteins, localizes to dynamic nuclear speckles; this localization pattern supports a role for the protein in pre-mRNA splicing. We found that human orthologs of UNC-75, whose splicing activity has recently been documented in vitro, are expressed nearly exclusively in brain and when expressed in C. elegans, rescue unc-75 mutant phenotypes and localize to subnuclear puncta. Furthermore, we report that the subnuclear-localized EXC-7 protein, the C. elegans ortholog of the neuron-restricted Drosophila ELAV splicing factor, acts in parallel to UNC-75 to also affect cholinergic synaptic transmission. In conclusion, we identified a new neuronal, putative pre-mRNA splicing factor, UNC-75, and show that UNC-75, as well as the C. elegans homolog of ELAV, is required for the fine tuning of synaptic transmission. These findings thus provide a novel molecular link between pre-mRNA splicing and presynaptic function. PMID- 12906793 TI - Cholesterol accumulation in NPC1-deficient neurons is ganglioside dependent. AB - Niemann-Pick type C (NPC) disease is a lysosomal disorder commonly caused by a recessive mutation in NPC1, which encodes an integral membrane protein with regions of homology to the morphogen receptor, Patched, and to 3-hydroxy-3 methylglutaryl coenzyme A reductase. Neurons in NPC disease exhibit extensive storage of free cholesterol and glycosphingolipids (GSLs), including GM2 and GM3 gangliosides. Most studies have viewed cholesterol storage as primary, with NPC1 functioning as a retroendocytic transporter for regulation of cholesterol homeostasis. Here, we analyze the effects of genetically depriving NPC neurons of complex gangliosides by creating mice doubly deficient in both NPC1 and the GSL synthetic enzyme, GM2/GD2 synthase (GalNAcT). Ganglioside and cholesterol expression in neurons of NPC1(-/-)/GalNAcT(+/+), NPC1(-/-)/GalNAcT(-/-), NPC1(+/+)/GalNAcT(-/-), and WT mice was examined in situ by immunocytochemical and histochemical methods. Neurons in double-deficient mice lacked intraneuronal GM2 accumulation as expected, but remarkably also exhibited absence or dramatic reduction in free cholesterol. Neurons storing cholesterol consistently showed GM3 accumulation but some GM3-positive neurons lacked cholesterol storage. These findings provide a compelling argument that cholesterol sequestration in NPC1 deficient neurons is ganglioside dependent and suggest that the function of NPC1 in these cells may be more closely linked to homeostatic control of GSLs than cholesterol. PMID- 12906794 TI - A conserved oligomerization domain in drosophila Bazooka/PAR-3 is important for apical localization and epithelial polarity. AB - The PAR-3/PAR-6/aPKC complex is required to establish polarity in many different cell types, including the C. elegans zygote and epithelial and neuronal cells in Drosophila and mammals. In each context, the components of this complex display a mutually dependent asymmetric cortical localization. PAR-6 is a direct effector of Rho family GTPases and binds to and regulates aPKC. Mammalian PAR-3 (mPar3) can associate with transmembrane proteins and may link the complex to the membrane, but this can account for only part of the requirement for this protein in the complex. Here we investigate the function of a novel conserved domain, CR1, of PAR-3 using computational, biochemical, and genetic approaches. Sequence structure comparison by FUGUE predicts that CR1 has the same structural fold as a bacterial oligomerization domain. We show that CR1 of the Drosophila homolog, Bazooka (BAZ), mediates oligomerization in vitro and in vivo. Furthermore, deletion of CR1 disrupts BAZ localization in both epithelial cells and the germline and strongly impairs BAZ function in epithelial polarity. These results indicate that this domain is important for the localization and activity of the PAR-3/PAR6/aPKC complex and define a new role for PAR-3 in assembling higher order protein complexes. PMID- 12906795 TI - The mouse Formin mDia1 is a potent actin nucleation factor regulated by autoinhibition. AB - Formin proteins are widely expressed in eukaryotes and play essential roles in assembling specific cellular actin-based structures. Formins are defined by a Formin Homology 2 (FH2) domain, as well as a proline-rich FH1 domain that binds the actin monomer binding protein, profilin, and other ligands. Constructs including FH2 of budding yeast Bni1 or fission yeast Cdc12 formins nucleate actin filaments in vitro. In this study, we demonstrate that FH2-containing constructs of murine mDia1 (also called p140 mDia or Drf1) are much more potent actin nucleators than the yeast formins. FH1 is necessary for nucleation when actin monomers are profilin bound. mDia1 is a member of the Diaphanous formin subfamily (Dia), whose members contain an N-terminal Rho GTPase binding domain (GBD) and a C-terminal Diaphanous autoinhibitory domain (DAD, ). Based on cellular and in vitro binding studies, an autoinhibitory model for Dia formin regulation proposes that GBD binding to DAD inhibits Dia-induced actin remodeling, whereas Rho binding activates by releasing GBD from DAD. Supporting this model, our results show that an N-terminal mDia1 construct strongly inhibits actin nucleation by the C terminus. RhoA partially relieves inhibition but does so when bound to either GDP or GTP analogs. Both N- and C-terminal mDia1 constructs appear to be multimeric. PMID- 12906796 TI - Requirements for Arabidopsis ATARP2 and ATARP3 during epidermal development. AB - Plant cells employ the actin cytoskeleton to stably position organelles, as tracks for long distance transport, and to reorganize the cytoplasm in response to developmental and environmental cues. While diverse classes of actin binding proteins have been implicated in growth control, the mechanisms of cytoskeletal reorganization and the cellular functions of specific actin filament arrays are unclear. Arabidopsis trichome morphogenesis includes distinct requirements for the microtubule and actin filament cytoskeletons. It also is a genetically tractable process that is providing new knowledge about cytoskeleton function in plants. The "distorted group" of mutants defines a class of at least eight genes that are required during the actin-dependent phase of trichome growth. Using map based cloning and a candidate gene approach, we identified mutations in ARP3 (ATARP3) and ARP2 (ATARP2) genes as the cause of the distorted1 (dis1) and wurm (wrm) phenotypes, respectively. ARP2 and ARP3 are components of the evolutionarily conserved ARP2/3 complex that nucleates actin filament polymerization [3]. Mutations in DIS1 and WRM caused severe trichome growth defects but had relatively mild effects on shoot development. DIS1 rescued the phenotype of Deltaarp3 when overexpressed in S. cerevisiae. Developing dis1 trichomes had defects in cytoplasmic actin bundle organization and reduced relative amounts of cytoplasmic actin filaments in developing branches. PMID- 12906797 TI - Learning and memory deficits in Notch mutant mice. AB - Notch is a critical component of evolutionarily conserved signaling mechanisms that regulate development and may contribute to plasticity-related processes, including changes in neurite structure and maintenance of neural stem cells. Deficits in the Notch pathway are responsible for Alagille and Cadasil syndromes, which are associated with mental retardation and dementia. Additionally, in postmitotic neurons, Notch proteins interact with presenilins and with beta amyloid precursor protein and could therefore have a role in the memory deficits associated with familial and sporadic Alzheimer's disease. To test if alterations in Notch signaling can lead to learning and memory deficits, we studied mice with mutations in this pathway. Here, we show that null heterozygous mutations in Notch1 result in deficits in spatial learning and memory without affecting other forms of learning, motor control, or exploratory activity. We also show that null heterozygous mutations in the downstream cofactor RBP-J result in similarly specific spatial learning and memory deficits. These data indicate that a constitutive decrease in Notch signaling can result in specific learning and memory deficits and suggest that abnormalities in Notch-dependent transcription may contribute to the cognitive deficits associated with Alzheimer's disease and Alagille and Cadasil syndromes. PMID- 12906799 TI - Planning for gene therapy. AB - In spite of some setbacks, gene therapy trials continue, while the UK government is putting in place plans to bring the potential power of genetics into mainstream public heathcare through its National Health Service. PMID- 12906800 TI - GM crops dancing to different tunes. AB - MEDIAWATCH: Parts of the British media are keeping up a campaign against the introduction of genetically modified crops ahead of the governemnet's assessment of the trial programme this autumn, often to the detriment of the arguments. PMID- 12906801 TI - Towering tribute to botany. AB - One of the world's greatest plant collections has won a top heritage award at a time when it is highlighting with a tree-top walkway the need to study the forest canopy which is one of the most crucial but least understood habitats. PMID- 12906802 TI - Obituary: Ira Herskowitz. AB - Jasper Rine pays tribute to a distinguished career spanning several fields and a personality that was a legend among the many great teachers at UCSF. PMID- 12906803 TI - Q & A. Philip Ingham. AB - Philip Ingham grew up in Liverpool and graduated from Cambridge University in 1977. He did his D. Phil in Developmental Genetics at Sussex University and postdoctoral work in Strasbourg, France before joining the laboratory of David Ish-Horowicz at the ICRF Mill Hill Laboratories. Here he applied the emerging technique of tissue in situ hybridisation to the analysis of the Drosophila segmentation genes. After a short spell at the MRC Laboratory of Molecular Biology in Cambridge, he rejoined the ICRF as a Research Scientist at the Developmental Biology Unit in Oxford. His group pioneered the analysis of the Hedgehog signalling pathway in Drosophila and in collaboration with the labs of Andy McMahon and Cliff Tabin at Harvard University, discovered the Hedgehog gene family in vertebrates. In 1996 he was appointed Professor of Developmental Genetics at the University of Sheffield where he has established the Centre for Developmental Genetics. PMID- 12906804 TI - Life in the fast lane. PMID- 12906805 TI - Low-fidelity DNA polymerases. PMID- 12906806 TI - Rhomboids. PMID- 12906807 TI - Evolution of the homeobox complex in the Diptera. PMID- 12906808 TI - Cytokinesis: a logical GAP. AB - Two complexes localize to the central spindle and regulate completion of cytokinesis: one, centralspindlin, contains a kinesin-like protein and a Rho family GAP; the second contains Aurora B kinase. Aurora B kinase is known to regulate localization of centralspindlin and may regulate the activity of the RhoGAP component of centralspindlin. PMID- 12906809 TI - Human evolution: a legacy of cannibalism in our genes? AB - A new study of genetic variation in the human prion protein gene suggests that balancing selection has operated on an amino acid sequence polymorphism in the gene during the last five hundred thousand years. Is this a legacy of widespread cannibalism by our ancestors? PMID- 12906810 TI - Replicative helicase loaders: ring breakers and ring makers. AB - Helicase loaders transfer the ring-shaped replicative helicases onto DNA. They assort into two classes: ring breakers, which place stabile hexameric rings on DNA via transient gaps at subunit interfaces; and helicase makers, which assemble hexameric rings around DNA from monomeric helicase units. PMID- 12906811 TI - Microtubule dynamics: new surprises from an old MAP. AB - Dis1/XMAP215 family microtubule-binding proteins are essential for cell division in animals, plants and fungi, suggesting a conserved cell-division mechanism used by all eukaryotes. Two new studies, however, reveal that different family members can have very different effects on microtubule dynamics. PMID- 12906812 TI - Spermatogenesis: borrowing the apoptotic machinery. AB - The apoptotic machinery is utilized for a wide variety of tasks during development. Recent work has uncovered a new, non-apoptotic role for these factors during the individualization process of maturing spermatids. PMID- 12906813 TI - Meiosis: polo, FEAR and the art of dividing reductionally. AB - Recent studies on the regulation of meiosis have uncovered new roles for old acquaintances: the polo-like kinase Cdc5 has been found to dictate proper kinetochore orientation during meiosis I, while the FEAR pathway is essential for some, but not all, aspects of meiosis I exit. PMID- 12906814 TI - Phage evolution: new worlds of genomic diversity. AB - A recent comparative survey of genomes of phages infecting mycobacteria reveals a vast combinatorial network of gene rearrangements and may provide general models for pattern and process in genome evolution. PMID- 12906815 TI - c-Abl regulation: a tail of two lipids. AB - c-Abl is a non-receptor tyrosine kinase whose activity is tightly controlled in vivo through unknown mechanisms. Recent studies suggest that c-Abl may be regulated in different cellular contexts by distinct lipids. PMID- 12906816 TI - Receptor-ligand binding: 'catch' bonds finally caught. AB - Leukocyte recruitment to sites of inflammation is initiated by the selectin family of adhesion receptors. Recent research reveals that P-selectin binding to its ligand exhibits 'catch' to 'slip' bond transition that may help explain the shear threshold phenomenon. PMID- 12906818 TI - The final player in the coenzyme A biosynthetic pathway. AB - The determination of the crystal structure of human phosphopantothenoylcysteine synthetase completes our knowledge of the enzyme structures involved in all steps of coenzyme A biosynthesis. This structure provides insight into the differences between bacterial and mammalian forms of the enzyme and may guide the structure based development of novel antibacterial compounds. PMID- 12906817 TI - The minus end in sight. AB - Microtubules are intrinsically polar structures. A consequence of this polarity is that the two ends of the microtubule polymer exhibit different properties. The more dynamic plus ends and the mechanisms that regulate their behavior have been the focus of much recent attention. Here, we concentrate on the dynamics and regulation of minus ends, which play distinct but equally critical roles in microtubule function. In the first part of this review, we compare the in vitro and in vivo behavior of microtubules from a minus end perspective. This comparison suggests that cells possess conserved mechanisms to specifically inhibit minus end polymerization, and perhaps also to actively promote depolymerization. In the second part, we focus on the spatial positioning of minus ends, which is achieved by localized microtubule nucleation, minus end capping and minus end anchoring as well as by motor-dependent sorting. These mechanisms are used in different biological contexts to generate the diversity of organized microtubule arrays in cells. PMID- 12906819 TI - Getting a grip on the CTD of Pol II. AB - The first structure of a pre-mRNA processing factor bound to heptad repeats from the C-terminal domain of RNA polymerase II is revealed in a crystal of capping guanylyltransferase complexed with a four-repeat phosphopeptide. PMID- 12906820 TI - Shikimate dehydrogenase structure reveals novel fold. AB - The structure of shikimate 5-dehydrogenase, the fourth enzyme in the shikimate biosynthesis pathway and a member of a large enzyme family without clear structural peer, reveals a novel topological fold for the substrate binding domain and, through homology modeling, expands the possibilities for antimicrobial and herbicide design. PMID- 12906821 TI - Sugar tongs get a grip on the starch granule in barley alpha-amylase 1. AB - A novel polysaccharide binding site is identified in domain C of barley alpha amylase 1 from the X-ray structure of the enzyme/tetrasaccharide complex. PMID- 12906822 TI - Coupling of folding and binding in the PTB domain of the signaling protein Shc. AB - The notion that certain proteins lack intrinsic globular structure under physiological conditions and that the attainment of fully folded structure only occurs upon the binding of target molecules has been recently gaining popularity. We report here the solution structure of the PTB domain of the signaling protein Shc in the free form. Comparison of this structure with that of the complex form, obtained previously with a phosphopeptide ligand, reveals that the Shc PTB domain is structurally disordered in the free form, particularly around the regions constituting the peptide binding pocket. The binding of the ligand appears to reorganize this pocket through local folding events triggering a conformational switch between the free and the complex forms. PMID- 12906823 TI - Structure and texture of fibrous crystals formed by Alzheimer's abeta(11-25) peptide fragment. AB - Amyloid fibril deposition is central to the pathology of Alzheimer's disease. X ray diffraction from amyloid fibrils formed from full-length Abeta(1-40) and from a shorter fragment, Abeta(11-25), have revealed cross-beta diffraction fingerprints. Magnetic alignment of Abeta(11-25) amyloid fibrils gave a distinctive X-ray diffraction texture, allowing interpretation of the diffraction data and a model of the arrangement of the peptides within the amyloid fiber specimen to be constructed. An intriguing feature of the structure of fibrillar Abeta(11-25) is that the beta sheets, of width 5.2 nm, stack by slipping relative to each other by the length of two amino acid units (0.70 nm) to form beta ribbons 4.42 nm in thickness. Abeta(1-40) amyloid fibrils likely consist of once folded hairpins, consistent with the size of the fibers obtained using electron microscopy and X-ray diffraction. PMID- 12906824 TI - Structure of human phosphopantothenoylcysteine synthetase at 2.3 A resolution. AB - The structure of human phosphopantothenoylcysteine (PPC) synthetase was determined at 2.3 A resolution. PPC synthetase is a dimer with identical monomers. Some features of the monomer fold resemble a group of NAD-dependent enzymes, while other features resemble the ribokinase fold. The ATP, phosphopantothenate, and cysteine binding sites were deduced from modeling studies. Highly conserved ATP binding residues include Gly43, Ser61, Gly63, Gly66, Phe230, and Asn258. Highly conserved phosphopantothenate binding residues include Asn59, Ala179, Ala180, and Asp183 from one monomer and Arg55' from the adjacent monomer. The structure predicts a ping pong mechanism with initial formation of an acyladenylate intermediate, followed by release of pyrophosphate and attack by cysteine to form the final products PPC and AMP. PMID- 12906825 TI - Structure of fully reduced bovine copper zinc superoxide dismutase at 1.15 A. AB - Copper zinc superoxide dismutase (CuZnSOD) forms a crucial component of the cellular response to oxidative stress by catalyzing the dismutation of the superoxide radical to hydrogen peroxide and water. Mutations in human CuZnSOD are associated with the development of familial amyotrophic lateral sclerosis (motor neuron disease). We have determined the structure of fully reduced bovine CuZnSOD to 1.15 A, the only atomic resolution structure for an intact CuZnSOD and one of only a small number for metalloproteins. For the first time, both subunits have been captured with the three coordinate Cu(I) ligation required by the generally accepted catalytic mechanism, where dismutation of the superoxide radical occurs via reduction of Cu. Furthermore, the improved resolution compared to previous studies (to 1.65 A) has allowed a more detailed examination of the metal center environment and its associated water network in the active site channel, facilitating the analysis of potential proton transfer routes. PMID- 12906826 TI - Structural basis of proline-specific exopeptidase activity as observed in human dipeptidyl peptidase-IV. AB - Inhibition of dipeptidyl peptidase IV (DPP-IV), the main glucagon-like peptide 1 (GLP1)-degrading enzyme, has been proposed for the treatment of type II diabetes. We expressed and purified the ectodomain of human DPP-IV in Pichia pastoris and determined the X-ray structure at 2.1 A resolution. The enzyme consists of two domains, the catalytic domain, with an alpha/beta hydrolase fold, and a beta propeller domain with an 8-fold repeat of a four-strand beta sheet motif. The beta propeller domain contributes two important functions to the molecule that have not been reported for such structures, an extra beta sheet motif that forms part of the dimerization interface and an additional short helix with a double Glu sequence motif. The Glu motif provides recognition and a binding site for the N terminus of the substrates, as revealed by the complex structure with diprotin A, a substrate with low turnover that is trapped in the tetrahedral intermediate of the reaction in the crystal. PMID- 12906827 TI - The 1.14 A crystal structure of yeast cytosine deaminase: evolution of nucleotide salvage enzymes and implications for genetic chemotherapy. AB - Cytosine deaminase (CD) catalyzes the deamination of cytosine and is only present in prokaryotes and fungi, where it is a member of the pyrimidine salvage pathway. The enzyme is of interest both for antimicrobial drug design and gene therapy applications against tumors. The structure of Saccharomyces cerevisiae CD has been determined in the presence and absence of a mechanism-based inhibitor, at 1.14 and 1.43 A resolution, respectively. The enzyme forms an alpha/beta fold similar to bacterial cytidine deaminase, but with no similarity to the alpha/beta barrel fold used by bacterial cytosine deaminase or mammalian adenosine deaminase. The structures observed for bacterial, fungal, and mammalian nucleic acid deaminases represent an example of the parallel evolution of two unique protein folds to carry out the same reaction on a diverse array of substrates. PMID- 12906828 TI - The structure of barley alpha-amylase isozyme 1 reveals a novel role of domain C in substrate recognition and binding: a pair of sugar tongs. AB - Though the three-dimensional structures of barley alpha-amylase isozymes AMY1 and AMY2 are very similar, they differ remarkably from each other in their affinity for Ca(2+) and when interacting with substrate analogs. A surface site recognizing maltooligosaccharides, not earlier reported for other alpha-amylases and probably associated with the different activity of AMY1 and AMY2 toward starch granules, has been identified. It is located in the C-terminal part of the enzyme and, thus, highlights a potential role of domain C. In order to scrutinize the possible biological significance of this domain in alpha-amylases, a thorough comparison of their three-dimensional structures was conducted. An additional role for an earlier-identified starch granule binding surface site is proposed, and a new calcium ion is reported. PMID- 12906829 TI - Structure of E. coli ketopantoate hydroxymethyl transferase complexed with ketopantoate and Mg2+, solved by locating 160 selenomethionine sites. AB - We report the crystal structure of E. coli ketopantoate hydroxymethyltransferase (KPHMT) at 1.9 A resolution, in complex with its product, ketopantoate. KPHMT catalyzes the first step in the biosynthesis of pantothenate (vitamin B(5)), the precursor of coenzyme A and the acyl carrier protein cofactor. The structure of the decameric enzyme was solved by multiwavelength anomalous dispersion to locate 160 selenomethionine sites and phase 560 kDa of protein, making it the largest structure solved by this approach. KPHMT adopts the (betaalpha)(8) barrel fold and is a member of the phosphoenolpyruvate/pyruvate superfamily. The active site contains a ketopantoate bidentately coordinated to Mg(2+). Similar binding is likely for the substrate, alpha-ketoisovalerate, orienting the C3 for deprotonation. PMID- 12906830 TI - A dual role for an aspartic acid in glycosylasparaginase autoproteolysis. AB - Glycosylasparaginase uses an autoproteolytic processing mechanism, through an N-O acyl shift, to generate a mature/active enzyme from a single-chain precursor. Structures of glycosylasparaginase precursors in complex with a glycine inhibitor have revealed the backbone in the immediate vicinity of the scissile peptide bond to be in a distorted trans conformation, which is believed to be the driving force for the N-O acyl shift to break the peptide bond. Here we report the effects of point mutation D151N. In addition to the loss of the base essential in autoproteolysis, this mutation also eradicates the backbone distortion near the scissile peptide bond. Binding of the glycine inhibitor to the autoproteolytic site of the D151N mutant does not restore the backbone distortion. Therefore, Asp151 plays a dual role, acting as the general base to activate the nucleophile and holding the distorted trans conformation that is critical for initiating an N O acyl shift. PMID- 12906831 TI - Crystal structure of shikimate 5-dehydrogenase (SDH) bound to NADP: insights into function and evolution. AB - The crystal structure of Methanococcus jannaschii shikimate 5-dehydrogenase (MjSDH) bound to the cofactor nicotinamide adenine dinucleotide phosphate (NADP) has been determined at 2.35 A resolution. Shikimate 5-dehydrogenase (SDH) is responsible for NADP-dependent catalysis of the fourth step in shikimate biosynthesis, which is essential for aromatic amino acid metabolism in bacteria, microbial eukaryotes, and plants. The structure of MjSDH is a compact alpha/beta sandwich with two distinct domains, responsible for binding substrate and the NADP cofactor, respectively. A phylogenetically conserved deep cleft on the protein surface corresponds to the enzyme active site. The structure reveals a topologically new domain fold within the N-terminal segment of the polypeptide chain, which binds substrate and supports dimerization. Insights gained from homology modeling and sequence/structure comparisons suggest that the SDHs represent a unique class of dehydrogenases. The structure provides a framework for further investigation to discover and develop novel inhibitors targeting this essential enzyme. PMID- 12906832 TI - Structure and mechanism of MT-ADPRase, a nudix hydrolase from Mycobacterium tuberculosis. AB - Nudix hydrolases are a family of proteins that contain the characteristic sequence GX(5)EX(7)REUXEEXG(I/L/V), the Nudix box. They catalyze the hydrolysis of a variety of nucleoside diphosphate derivatives such as ADP-ribose, Ap(n)A (3 100-fold in poor vector/parasite combinations (P. gallinaceum/An. stephensi), to complete blockade (>1,500 fold) in others (P. berghei/Ae. aegypti). Plasmodium berghei ookinetes survive poorly in the bloodmeal of Ae. aegypti and are unable to invade the midgut epithelium. Cultured mature ookinetes of P. berghei injected directly into the mosquito haemocoele produced salivary gland sporozoites in An. stephensi, but not in Ae. aegypti, suggesting that further species-specific incompatibilities occur downstream of the midgut epithelium in Ae. aegypti. These results show that in these parasite mosquito combinations the susceptibility to malarial infection is regulated at multiple steps during the development of the parasites. Understanding these at the molecular level may contribute to the development of rational strategies to reduce the vector competence of malarial vectors. PMID- 12906878 TI - Echinococcus multilocularis: secondary poisoning of fox population during a vole outbreak reduces environmental contamination in a high endemicity area. AB - This paper describes the role of fox population level on Echinococcus multilocularis infection in foxes in a highly endemic area in eastern France. Fox population level was monitored by spotlight survey at Le Souillot from 1989 to 2000, and from 1992 to 2000 at Chemin, a control site located in a low endemic area. The infection level of the fox population was estimated at Le Souillot from winter 1995 to winter 1999 using a coproantigen ELISA performed on faeces collected in the field. Population biomass of intermediate hosts (Microtus arvalis and Arvicola terrestris) was monitored using index methods from 1995 to 1999. At Le Souillot, a significant decline in the fox population level was recorded after spring 1997 (P<0.001), and the population level remained low until 2000. The decline occurred when 31% of the grassland area was treated with bromadiolone, an anticoagulant used at a large scale for the control of A. terrestris population outbreaks. No decline of population was recorded at Chemin, where bromadiolone was not used for rodent control. Significant differences among ELISA OD distributions in fox faeces were recorded for the five winters under study at Le Souillot (P=0.0004). The median of ELISA OD distribution was 0.209 and 0.207 before the population decline (winter 1995 and 1996, respectively), significantly increased to 0.306 just after the decline (winter 1997), and then significantly decreased to 0.099 and 0.104 afterwards (winter 1998 and 1999, respectively). Therefore, the decrease in infection level occurred during winter 1998, 1 year after the population decline, when the intermediate host biomass in the field was at its highest. These results suggest a complex dependence between the fox population level and E. multilocularis infection in a high endemicity area. Alternative ways to control fox population as a way to reduce E. multilocularis transmission in a given area are discussed. PMID- 12906879 TI - Comparative analysis of mitochondrial genome data for Necator americanus from two endemic regions reveals substantial genetic variation. AB - Necator americanus is a blood-sucking, intestinal nematode of major human health importance in many tropical and subtropical regions of the world. The aim of the present study was to compare the complete mitochondrial genome sequence from one N. americanus individual from Togo with another from China, in order to estimate the magnitude of genetic variability for different mitochondrial genes and non coding regions. For the 12 protein genes, this comparison revealed sequence differences at both the nucleotide (3-7%) and amino acid (1-7%) levels. The most conserved of these was the nad4L gene, whereas the nad1 gene was least conserved at both the nucleotide and amino acid levels. Nucleotide differences were also detected in 14 of the 22 transfer RNAs (trns) (1-13%), the AT-rich region ( approximately 8%), non-coding regions (8-25%) and in the small (rrnS) and large (rrnL) subunits of mitochondrial ribosomal RNA (rrn) ( approximately 1%). Comparison of the rrnL sequences among multiple individual worms revealed nine unequivocal nucleotide differences between N. americanus from the two countries. Consistent with previous studies, these findings provide evidence for substantial genetic variation within N. americanus, which may have implications for the transmission and control of hookworm disease. PMID- 12906880 TI - Sense and antisense transcripts in the histone H1 (HIS-1) locus of Leishmania major. AB - Histone H1 in the parasitic protozoan Leishmania is a developmentally regulated protein encoded by two genes, HIS-1.1 and HIS-1.2. These genes are separated by approximately 20 kb of sequence and are located on the same DNA strand of chromosome 27. When Northern blots of parasite RNA were probed with HIS-1 strand specific riboprobes, we detected sense and antisense transcripts that were polyadenylated and developmentally regulated. When the HIS-1.2 coding region was replaced with the coding region of the neomycin phosphotransferase gene, antisense transcription of this gene was unaffected, indicating that the regulatory elements controlling antisense transcription were located outside of the HIS-1.2 gene, and that transcription in Leishmania can occur from both DNA strands even in the presence of transcription of a selectable marker in the complementary strand. A search for other antisense transcripts within the HIS-1 locus identified an additional transcript (SC-1) within the intervening HIS-1 sequence, downstream of adenine and thymine-rich sequences. These results show that gene expression in Leishmania is not only regulated polycistronically from the sense strand of genomic DNA, but that the complementary strand of DNA also contains sequences that could drive expression of open reading frames from the antisense strand of DNA. These findings suggest that the parasite has evolved in such a way as to maximise the transcription of its genome, a mechanism that might be important for it to maintain virulence. PMID- 12906881 TI - A newly-identified lineage of Schistosoma. AB - Because of their role in causing schistosomiasis, flukes of the genus Schistosoma are the best known of all digeneans. The genus has traditionally been divided into four familiar species groups. Here we report on three poorly known species of Schistosoma, one of which, Schistosoma hippopotami, is known from the hippopotamus, one of which is provisionally identified as Schistosoma edwardiense, another hippo parasite, and a third that has not previously been described. All were collected from freshwater snails obtained from Lake Edward, western Uganda, the type locality for both known hippo schistosomes. The three different kinds of schistosome cercariae differ from one another in size, and all are readily differentiated by their long tail stems from the cercariae of human infecting species. Furthermore, each was recovered from a different genus of snail host, Biomphalaria sudanica, Bulinus truncatus or Ceratophallus natalensis. Molecular analysis, based on 8350 bases of combined nuclear and mitochondrial DNA, groups these three long tail-stem cercariae into a well supported clade that does not associate with any of the recognised species groups. The placement of this clade, basal to all African species plus several Asian species, suggests that there has been an ancient association between Schistosoma and hippos. This new African Schistosoma clade advocates the need for further modification of the traditional species group-based classification. Two of the four species groups are paraphyletic. It also suggests that Schistosoma has been remarkably plastic with respect to adapting to snail hosts-three distantly related genera of planorbid snails have been exploited by worms within a single clade. Finally, it adds a new layer of complexity to deciphering the origins of Schistosoma, often considered to be African but recently challenged as being Asian. In the late Cenozoic the distribution of hippo species straddled both Africa and Asia and they may have provided a means for the introduction of blood flukes to Africa. PMID- 12906882 TI - Cyclosporin-binding proteins of Plasmodium falciparum. AB - A number of cyclosporins, including certain non-immunosuppressive ones, are potent inhibitors of the intraerythrocytic growth of the human malarial parasite Plasmodium falciparum. The major cyclosporin-binding proteins of P. falciparum were investigated by affinity chromatography on cyclosporin-Affigel followed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis, Western blotting, and peptide mass fingerprinting. The two bands obtained on gels were shown to correspond to cyclophilins, PfCyP-19A (formerly PfCyP-19) and PfCyP-19B, whose genes had been characterised previously. PfCyP-19B was an abundant protein of intraerythrocytic P. falciparum (up to 0.5% of parasite protein) that was present in the highest amounts in schizont-stage parasites. Unexpectedly, given its apparent signal sequence, it was located primarily in the cytosol of the parasite. The peptidyl-prolyl cis-trans isomerase activity of recombinant PfCyP 19B had the same profile of susceptibility to cyclosporin derivatives as the bulk isomerase activity of crude P. falciparum extracts. The binding of cyclosporins to cyclophilins may be relevant to the mechanism of action of the drug in the parasite. PMID- 12906883 TI - Animal experimentation in snake venom research and in vitro alternatives. AB - Current experimental techniques used in snake venom research (with and without the use of animals) are reviewed. The emphasis is on the reduction of the use of animals in the development of antivenoms for the clinical treatment of snakebite. Diagnostic and research techniques for the major pathologies of envenoming are described and those using animals are contrasted with non-sentient methods where possible. In particular, LD50 and ED50 assays using animals (in vivo) and fertilised eggs (in vivo, non-sentient) are compared as well as in vitro procedures (ELISA and haemolytic test) for ED50 estimations. The social context of antivenom production, supply and demand is outlined together with the consequent tension between the benefits derived and the increase in opposition to experiments on animals. Stringent regulations governing the use of animals, limited research funds and public pressure all focus the need for progress towards non-animal, or non-sentient, research methods. Some achievements are noted but success is hampered by lack of detailed knowledge of the many constituents of venom which have to be assessed as a whole rather than individually. The only way to evaluate the net pathological effect of venom is to use a living system, usually a rodent, and similarly, the efficacy of antivenoms is also measured in vivo. The pre-clinical testing of antivenoms in animals is therefore a legal requirement in many countries and is strictly monitored by government authorities. New technologies applied to the characterisation of individual venom proteins should enable novel in vitro assays to be designed thus reducing the number of animals required. In the meantime, the principles of Reduce, Refine and Replace relating to animals in research are increasingly endorsed by those working in the field and the many agencies regulating ethical and research policy. PMID- 12906884 TI - A potential vector of domoic acid: the swimming crab Polybius henslowii Leach (Decapoda-brachyura). AB - The swimming crab Polybius henslowii may play an important role in the movement of the amnesic shellfish toxin, domoic acid (DA), through the marine food chain. High DA concentrations have been determined in crab samples harvested along the Portuguese coast during the summer of 2002, reaching a level of 323.1 microg DA/g crab tissue. Toxin distribution in the different crab organs showed levels as high as 571.6 microg DA/g in the visceral tissues. Levels of toxin 4-12 times lower were detected in the remaining tissues. This crab might be a prominent vector of the toxin to higher trophic levels, including fishes, sea birds and even humans. In Portugal P. henslowii is commercialised during the summer in some local markets. DA concentrations were found close to the legal limit of 20 microg/g in samples purchased at Figueira da Foz market. The crabs are boiled prior to reaching the consumers. The cooking process was evaluated. Determination of toxin losses during the cooking process showed a toxin reduction higher than 50%. DA was determined by HPLC-UV and confirmed by spectra acquired with diode array detector. PMID- 12906885 TI - Inhibitory potential of Crotalus durissus terrificus venom on measles virus growth. AB - This paper presents the antiviral activity found in a snake with Crotalus durissus terrificus venom (Cdt), studied by use of microplate inhibition assay, using measles virus (MV). Cdt at concentrations below 100 microg/ml showed no cytotoxicity for Vero cells. This study shows the optimal conditions for cell treatment and infection. Two factors that affect virus binding and infection efficiency were studied: the use of an adsorption step, where infection volume was varied; and the concentration of fetal bovine serum (FBS). The adsorption step, with or without FBS, increased the bound virus percentage, whereas it increased bound virus at equilibrium only in FBS-free until 2.5% FBS. In contrast, the addition of 10% FBS decreased the bound virus percentage. The inhibition of MV replication in Vero cells was observed when Cdt was added either before or during cell infection with virus. Its inhibitory concentration against MV replication was 0.1 until 100 microg/ml, respectively. The anti-MV effect of the Cdt was gradually decreased when it was added before or during infection, and little inhibition was observed when Cdt was added 1 h after infection, suggesting that the MV infection was inhibited at the time of the initial events such as at the moment of adsorption and penetration of the viral cycle. In conclusion, Cdt contains anti-MV effects that may be of potential clinical interest. PMID- 12906886 TI - Hematological cell findings in bone marrow and peripheral blood of rabbits after experimental acute exposure to Loxosceles intermedia (brown spider) venom. AB - The purpose of this work was to find out the cellular changes occurring in bone marrow and peripheral blood after acute exposure to the venom of Loxosceles intermedia. Doses of 40 microg of venom were injected intradermally into five rabbits, and five rabbits receiving only phosphate-buffered saline (PBS) were used as controls. Bone marrow and peripheral blood samples were obtained before the envenomation and 4, 8, 12, 24 and 48 h, and 5, 10, 15, 20 and 30 days after envenomation. In bone marrow samples we assessed cellularity, nucleated red cells, megakaryocytes and neutrophils, and in peripheral blood we assessed red cells (red cell concentration, hemoglobin and hematocrit), leukocytes, neutrophils and platelets. Our objective was to find out if the venom has a direct effect on bone marrow and peripheral blood or if changes in both of them are secondary to the needs of tissues, and if there is a good correlation between histopathological and hematological findings. We found that the red cell parameters were not affected by the venom, except for nucleated red cells which decreased after venom exposure. The depression of megakaryocyte numbers and thrombocytopenia showed a strong correlation with the histopathologic changes observed in skin biopsies obtained from the rabbits. The changes in cellularity and neutrophils of bone marrow were strongly correlated with those in peripheral blood and skin. The thrombocytopenia and neutropenia in peripheral blood are due to marrow depression, which may be a consequence of an extensive migration of platelets and neutrophils to the necrotic lesion or the marrow depression may be a transitory effect of evenoming by L. intermedia. PMID- 12906887 TI - A prospective study of definite bites by spiders of the family Sparassidae (huntsmen spiders) with identification to species level. AB - Spiders of the family Sparassidae occur on most continents in tropical and temperate regions of the world. They are large Araneomorphae (modern spiders) which are often feared. There are few reports of confirmed sparassid bites and some suggest that particular genera (Neosparassus) can cause severe effects. This study investigated the circumstances and clinical effects of bites by Australian sparassid spiders with correlation to taxonomic species level. Cases were recruited prospectively from calls to Australian poison information centres and two emergency departments. Patients were included if there was a clear history of bite and the spider was caught, and were followed up over a week. There were 173 patients with spider injuries and 168 with definite bites by sparassid spiders over a 27 month period. Ninety-five percent of bites were by spiders from four genera: Isopeda (32%), Isopedella (21%), Neosparassus (27%) and Heteropoda (14%); other genera included Delena and Holconia. The seasonal and diurnal distribution of bites differed between genera, although the majority occurred in warmer months in daylight hours. Seventy-six percent of bites occurred because the spider was interfered with (picked up, dressing or picked up an object with the spider) consistent with the behaviour of most of the family requiring provocation to bite. Ninety-five percent of bites occurred on limbs, 82% on distal limbs, again consistent with handling the spider. Pain/discomfort occurred in all cases, and was severe in 27%. The median duration of pain was 5 min which was significantly less than other spiders. Puncture marks (40%) or localised initial bleeding (35%) occurred in 54% of bites, and both occurred significantly more often in all sparassid bites compared to all other spider bites (p<0.0001). Swelling occurred in 16%, itchiness in 14% and redness/red mark in 57% of cases, the latter significantly less common than other spiders (p=0.0002). Systemic effects occurred in 4% of bites and were minor (nausea and/or headache). There were no cases of necrotic ulcers or allergic reactions. There were four cases with features consistent with local infection. Bites by sparassid spiders cause minor effects, characterised by immediate and transient pain, associated with bleeding, puncture marks and local redness. The mechanism of effects appeared to be trauma rather than envenoming. There were no major differences between genera, and Neosparassus spp. did not cause major effects. PMID- 12906888 TI - Inhibition of hemorrhagic and edematogenic activities of snake venoms by a broad spectrum protease inhibitor, murinoglobulin; the effect on venoms from five different genera in Viperidae family. AB - In order to obtain basic data on the effect of broad-spectrum protease inhibitor against local symptoms of Viperidae snake envenomation, inhibitory capacity of rat murinoglobulin on local hemorrhagic and edematogenic activities of venoms from Crotalus atrox, Bothrops jararaca, Lachesis muta muta, Trimeresurus flavoviridis and Echis carinatus sochureki were examined. Murinoglobulin, pre incubated with the crude venoms at 37 degrees C for 15 min, inhibited hemorrhagic activity of all five venoms to various extents. The activity of C. atrox was almost completely inhibited at the murinoglobulin/venom ratio (w/w) of 20. The activity of B. jararaca, Lachesis muta muta and T. flavoviridis venoms was considerably inhibited at the ratio of 20 (77.2, 80.0 and 86.2% inhibition, respectively), however some of the activity still remained even at the ratio of 40 (84.2, 79.8 and 86.2% inhibition, respectively). Among the five venoms, E. c. sochureki venom is quite resistant to murinoglobulin treatment and statistically significant inhibition was only found at the ratio of 40 (64.1% inhibition). Fibrinolytic and gelatinase activities were more susceptible to murinoglobulin inhibition. The treatment at the ratios of 10 and 20 almost completely inhibited respectively the fibrinolytic and the gelatinase activities of all the venoms. Murinoglobulin treatment also significantly inhibited the edematogenic activity of L. muta muta, T. flavoviridis and Echis carinatus sochureki. The treatment of murinoglobulin at the ratio of 40 considerably suppressed the swelling up to 60 min after subcutaneous injection of L. muta muta and E. c. sochureki venoms, and up to 30 min after T. flavoviridis venom injection. Murinoglobulin is a potent inhibitor against local effects of multiple snake venoms in Viperidae family. PMID- 12906889 TI - Role of mast cells and pro-inflammatory mediators on the intestinal secretion induced by cholera toxin. AB - Recent data suggest that diarrhea caused by Vibrio cholerae involves a pro inflammatory mediators release, such as cytokines, prostaglandin and nitric oxide. The aim of this study was to investigate the role of mast cells and their mediators in the intestinal secretion induced by cholera toxin. We examined the dose responses, time course and role of mast cells and pro-inflammatory mediators in cholera toxin intestinal secretory response, in vivo. Cholera toxin caused a dose-dependent secretion, in ligated small intestine loops, at 18 h. Rats treated with 48/80 compound or ketotifen had a significant decrease in the intestinal secretory response. Cholera toxin secretion was significantly reduced by an unspecific histamine/serotonin receptor antagonist, histamine receptor antagonist, phospholipase A2 and cyclooxygenase inhibitors, platelet-activating factor (PAF) receptor antagonists and TNF-alpha synthesis blockers. On the other hand, pretreatment with a specific serotonin receptor antagonist and lipoxygenase inhibitors failed to block this effect. Analysis of the intestinal fluid from rats injected with cholera toxin, revealed that cholera toxin induces the release of IL-1beta and TNF-alpha into fluid. The data suggest that, at least in part, mast cells are involved in cholera toxin-induced secretion, as well as point to the importance of histamine, prostaglandins, PAF, IL-1beta and TNF-alpha in this process. PMID- 12906890 TI - A rapid assay for the brevetoxin group of sodium channel activators based on fluorescence monitoring of synaptoneurosomal membrane potential. AB - A functional pharmacologically-based assay for the brevetoxin group of sodium channel activators was developed using synaptoneurosomes isolated from the brains of CD1 mice. The assay can detect the depolarizing effect of brevetoxin congeners PbTx-2 and PbTx-3 as enhancements of the veratridine-dependent increase in fluorescence of the voltage-sensitive fluorescent probe rhodamine 6G. The assay is relatively rapid and can detect brevetoxin activity in the nanomolar range. The synaptoneurosomal assay has been used to analyse mussel tissue extracts spiked with PbTx-2, and composite toxicity, expressed as PbTx-3 equivalents in extracts of oysters naturally exposed to brevetoxins. In this latter context, the synaptoneurosomal technique was shown to compare favorably with the cytotoxicity assay, the receptor binding assay and HPLC/MS. Our results support the concept that this membrane potential assay detects brevetoxins based on their interaction with sodium channels. PMID- 12906891 TI - BmKK4, a novel toxin from the venom of Asian scorpion Buthus martensi Karsch, inhibits potassium currents in rat hippocampal neurons in vitro. AB - A novel short-chain peptide BmKK4 was isolated from the venom of Asian scorpion Buthus martensi Karsch. It is composed of 30 amino acids including six cysteine residues, and shares less than 25% sequence identity with the known alpha-KTx toxins. The action of BmKK4 on voltage-dependent potassium currents was examined in acutely dissociated hippocampal neurons of rat. BmKK4 (10-100 microM) inhibited both the delayed rectifier and fast transient potassium current in concentration-dependent manners. The inhibition was reversible and voltage independent. BmKK4 caused a depolarizing shift (about 10 mV) of the steady-state activation curve of the currents, without changing their steady-state inactivation behavior. The unique amino acid sequence and electrophysiological effects suggest that BmKK4 represent a new subfamily of potassium channel toxins. PMID- 12906892 TI - Fractionation and characterization of Cerastes cerastes cerastes snake venom and the antitumor action of its lethal and non-lethal fractions. AB - In the present study fractionation of the Cerastes cerastes cerastes snake venom by gel filtration on Sephadex G-75 gave 14 protein fractions. Phospholipase PLA2 activity is not uniformly correlated with the lethality to mice in regard to all venom fractions. F11 which is the richest in PLA2 activity is less toxic than F3, which contains a small amount of PLA2, and F12 is the lowest in lethality and PLA2 activity. Treatment of Ehrlich ascites-bearing mice with two i.p. injections of the most lethal fraction (F3) or a non-lethal fraction (F4) resulted in a significant antitumor activity demonstrated by an increase in the mean survival time of the animals (22.5 and 27.9 days) and in the tumor inhibition ratio of tumor growth (T/C% 139 and 172, respectively), compared to tumor-bearing controls. The cytotoxic activity of F3 and F4 against Ehrlich ascites carcinoma cells might be due to the presence of a cytotoxin rather than to the direct cytolytic effect of the PLA2 because the non-lethal F4 is free from PLA2. Treatment of Swiss albino mice with two i.p. injections of F3 or F4 at the adopted dose levels produced no detrimental side effects demonstrated by the insignificant changes in the tested serum and liver parameters. Treatment of the tumor-bearing mice with the same venom fractions significantly modulated all of the studied biochemical parameters in the serum and liver tissues, compared to normal controls. PMID- 12906893 TI - A simple technique for milking the venom of a small parasitic wasp, Melittobia digitata (Hymenoptera: Eulophidae). AB - We describe a simple method for obtaining good a quantity of pure venom from a small parasitoid wasp, Melittobia digitata. Crushing the insect's head causes venom to be extruded from the ovipositor that dries rapidly as it is collected onto an insect pin. This technique may be applicable to other parasitic Hymenoptera. PMID- 12906895 TI - Analytical methods for the quantitative determination of 3-hydroxy-3 methylglutaryl coenzyme A reductase inhibitors in biological samples. AB - Published analytical methods for the quantitative determinations of presently available five 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors ("statins"), lovastatin, simvastatin, pravastatin, fluvastatin and atorvastatin, are reviewed for therapeutic drug monitoring purpose in patients. Almost all assay reviewed are based on high-performance liquid chromatography or gas chromatography. Some purification steps (liquid-liquid extraction, solid-phase extraction, etc.) have been used before they are submitted to separation by chromatographic procedures and they are detected by various detection methods like UV, fluorescence and mass spectrometry. This review shows that most method may be used quantitative determination of statins in plasma and they are suitable for therapeutic drug monitoring purpose of these drugs. PMID- 12906896 TI - Enantioselective quantitation of the ecstasy compound (R)- and (S)-N-ethyl-3,4 methylenedioxyamphetamine and its major metabolites in human plasma and urine. AB - An enantioselective HPLC method has been developed and validated for the stereospecific analysis of N-ethyl-3,4-methylenedioxyamphetamine (MDE) and its major metabolites N-ethyl-4-hydroxy-3-methoxyamphetamine (HME) and 3,4 methylenedioxyamphetamine (MDA). These compounds have been analyzed both from human plasma and urine after administration of 70 mg pure MDE-hydrochloride enantiomers to four subjects. The samples were prepared by hydrolysis of the o glucuronate and sulfate conjugates using beta-glucuronidase/arylsulfatase and solid-phase extraction with a cation-exchange phase. A chiral stationary protein phase (chiral-CBH) was used for the stereoselective determination of MDE, HME and MDA in a single HPLC run using sodium dihydrogenphosphate, ethylendiaminetetraacetic acid disodium salt and isopropanol as the mobile phase (pH 6.44) and fluorimetric detection (lambda(ex) 286 nm, lambda(em) 322 nm). Moreover, a suitable internal standard (N-ethyl-3,4-methylenedioxybenzylamine) was synthesized and qualified for quantitation purposes. The method showed high recovery rates (>95%) and limits of quantitation for MDE and MDA of 5 ng/ml and for HME of 10 ng/ml. The RSDs for all working ranges of MDE, MDA and HME in plasma and urine, respectively, were less than 1.5%. After validation of the analytical methods in plasma and urine samples pharmacokinetic parameters were calculated. The plasma concentrations of (R)-MDE exceeded those of the S enantiomer (ratio R:S of the area under the curve, 3.1) and the plasma half time of (R)-MDE was longer than that of (S)-MDE (7.9 vs. 4.0 h). In contrast, the stereochemical disposition of the MDE metabolites HME and MDA was reversed. Concentrations of the (S)-metabolites in plasma of volunteers were much higher than those of the (R)-enantiomers. PMID- 12906897 TI - Determination of urinary hippuric acid by micellar electrokinetic capillary chromatography. AB - We propose a method for the simultaneous determination of hippuric acid (HA) and creatinine based on capillary micellar electrokinetic chromatography. Experimental conditions were 20 mM sodium phosphate, pH 7.20, 25 mM sodium dodecyl sulfate, 5% (v/v) acetonitrile. Electropherograms evidenced HA and creatinine peaks in less than 12 min. The method showed good linearity for both analytes and satisfactory within-day precision. The present method, which is accurate, sensitive, rapid and simple, may be applied to single-spot urine samples. PMID- 12906898 TI - Imaging surface plasmon resonance system for screening affinity ligands. AB - A surface plasmon resonance (SPR) system for screening ligands for application in affinity chromatography is described. A combinatorial library of 13 ligands was synthesised, characterised and immobilised to agarose beads and gold SPR devices. Binding and elution behaviour and a range of K(AX) values (10(3) to 10(5) M(-1)) were measured against two target proteins, an insulin analogue (MI3) and a recombinant clotting factor (rFVIIa), in order to create a relational database between the traditional chromatographic format and the new SPR screening system. The SPR transducer surface was fabricated with affinity ligands in a two dimensional, spatially addressable format, which was durable (>100 cycles) and stable over 6 months. The imaging SPR system comprised a direct optical, CCD based, instrument capable of imaging the change in refractive index created by biochemical interactions and allowed affinity ligands to be evaluated 15-fold faster with 130-fold less target protein than conventional chromatographic methods. The binding and elution data from both the SPR and chromatographic systems for both target proteins were comparable, with the K(AX) value generating a nearly linear correlation (R(2)=0.875) and a slope bias of approximately 2.5+/ 0.25-fold higher for the SPR system. The imaging SPR system has proven capable of screening and evaluating affinity ligands for potential use in the recovery of biopharmaceutical proteins. PMID- 12906899 TI - Liquid chromatography-tandem mass spectrometric assay with a novel method of quantitation for the simultaneous determination of bulaquine and its metabolite, primaquine, in monkey plasma. AB - A sensitive and selective liquid chromatography-tandem mass spectrometry method (LC-MS-MS) for the simultaneous estimation of bulaquine and primaquine has been developed and validated in monkey plasma. The mobile phase consisted of acetonitrile/ammonium acetate buffer (20 mM, pH 6) (50:50 v/v) at a flow-rate of 1 ml/min. The chromatographic separations were achieved on two spheri cyano columns (5 microm, 30 x 4.6 mm I.D.) connected in series. The quantitation was carried out using a Micromass LC-MS-MS with an electrospray source in the multiple reaction monitoring (MRM) mode. The analytes were quantified from the summed total ion value of their two most intense molecular transitions. This is another novel method leading to increased sensitivity and precision. A simple liquid-liquid extraction with 2 x 1.0 ml n-hexane/ethyl acetate/dimethyloctyl amine (90:10:0.05, v/v) was utilized. The method was validated in terms of recovery, linearity, accuracy and precision (within- and between-assay variation). The recoveries from spiked control samples were >or=90 and 50% for bulaquine and primaquine, respectively. Linearity in plasma was observed over a dynamic range of 1.56-400 and 3.91-1000 ng/ml for bulaquine and primaquine, respectively. PMID- 12906900 TI - Estimation of octanol-water partition coefficients for neutral and weakly acidic compounds by microemulsion electrokinetic chromatography using dynamically coated capillary columns. AB - Microemulsion electrokinetic chromatography (MEEKC) using dynamically coated capillary columns is shown to be suitable for estimating the octanol-water partition coefficient (log P) for neutral and weakly acidic compounds at pH 3. The solvation parameter model is used to demonstrate that the retention properties of sodium dodecyl sulfate (1.4% w/v), n-butanol (8% v/v) and n-heptane (1.2% v/v) microemulsion are strongly correlated with the octanol-water partition system. For compounds of varied structure and log P values from 0.3 to 5.15, the correlation model is able to estimate log P to better than 0.25 log units. The dynamically coated columns consisting of a bilayer of poly(vinylsulfonate) adsorbed on top of polybrene provide a suitable electroosmotic flow at pH 3 without interfering in the retention properties of the microemulsion. For automated measurements the microemulsion run buffer should be replenished after 10 runs to maintain a stable cycle time and the coated columns replaced after 40 70 runs, depending on sample properties. PMID- 12906901 TI - Purification of single-strand DNA binding protein from an Escherichia coli lysate using counter-current chromatography, partition and precipitation. AB - Single-strand DNA binding protein (SSB) from Escherichia coli lysate was purified by counter-current chromatography (CCC) using the ammonium sulfate precipitation method in a coiled column. About 5 ml of E. coli lysate was separated by CCC using a polymer phase system composed of 16% (w/w) polyethylene glycol (PEG) 1000 and 17% (w/w) ammonium sulfate aqueous polymer two-phase solvent system. The precipitation of proteins in the lysate took place in the CCC column, and the SSB protein was eluted in the fraction 51-56. Many other impurities were either eluted immediately after the solvent front or precipitated in the column. The identities of the proteins in the fractions and in the precipitate were confirmed by SDS-polyacrylamide gel electrophoresis with Coomassie Brilliant Blue staining. PMID- 12906902 TI - Rapid and sensitive quantitation of the antiproliferative agent mitoguazone in small volumes of plasma by high-performance liquid chromatography with ultraviolet detection. AB - Mitoguazone is an antiproliferative agent used in chemotherapy. This study describes a simple and sensitive high-performance liquid chromatographic method for the determination of mitoguazone in 100 microl of plasma. Samples were deproteinized with 100 microl of a solution of internal standard (amiloride, 10 microg/ml) in acetonitrile. An aliquot of the supernatant was injected onto the column. HPLC separation was achieved on a silica column with the mobile phase of methanol-50 mM potassium phosphate buffer (pH 3)-triethylamine (80:20:0.3, v/v), at a flow-rate of 1 ml/min. The eluent was detected at 320 nm. The retention time was about 5.5 min for amiloride and 12 min for mitoguazone. No endogenous substances were found to interfere. Calibration curves were linear from 0.25 to 50 microg/ml. The absolute recoveries of mitoguazone and amiloride were both greater than 84%. The limit of quantitation was 0.25 microg/ml. The intra- and inter-day precision (expressed as RSD) was 5.8%, or less, and the accuracy was 94.7% of the nominal concentration. The method is suitable in pharmacokinetic investigation and monitoring mitoguazone concentration. PMID- 12906903 TI - High-performance liquid chromatographic method for the determination of (-) verbenone 10-hydroxylation catalyzed by rat liver microsomes. AB - A sensitive assay for the determination of (-)-verbenone 10-hydroxylation catalyzed by rat liver microsomes was developed using high-performance liquid chromatography. Verbenone was incubated in vitro with liver microsomes of untreated rats and rats treated with phenobarbital and the products thus formed were extracted with CH(2)Cl(2) and the extracts were separated by HPLC with a C(18) 5-microm analytical column. Elution was conducted with 40% methanol containing 20 mM NaClO(4) and the detection of UV absorbance was done at 251 nm. Product formation was dependent on the incubation time at least up to 30 min and the microsomal protein concentration between 0.01 and 0.1 mg protein/ml. The limit of detection of (-)-10-hydroxyverbenone with the HPLC was found to be about 40 pg, indicating that this method is about 100-fold sensitive than the GC-MS method. Optimized pH for the reaction was at 7.4 when examined with 100 mM potassium phosphate buffer in different pHs. Kinetic analysis showed that K(m) values for liver microsomes of untreated and phenobarbital-treated rats were 206 and 41 microM and V(max) values were 5.8 and 44 nmol/min/mg protein, respectively. Thus the present results provided a sensitive and useful method for the determination of verbenone 10-hydroxylation catalyzed by rat liver microsomes. PMID- 12906904 TI - Liquid chromatographic-tandem mass spectrometric method for the plant lignan 7 hydroxymatairesinol and its potential metabolites in human plasma. AB - A HPLC-MS-MS method was developed for the determination of the plant lignan 7 hydroxymatairesinol and its potential metabolites matairesinol, oxomatairesinol, alpha-conidendrin, 7-hydroxyenterolactone, enterodiol, and enterolactone in human plasma. The method included sample cleanup by solid-phase extraction (SPE) and analysis using a PE Sciex API3000 triple quadrupole mass spectrometer with electrospray ionisation. The lignans were quantified using two deuterated internal standards. They showed good chromatographic linearity, analysis repeatability, and SPE recovery in the presence of plasma. In pooled plasma and in plasma samples collected from two individual subjects lignan glucuronides and sulfates were enzymatically hydrolysed to free lignans and then analysed. All the lignans could be detected in the samples. PMID- 12906905 TI - Determination using liquid-chromatography-electrospray tandem mass spectroscopy of ethinylestradiol serum pharmacokinetics in adult Sprague-Dawley rats. AB - The pharmacokinetics of ethinylestradiol (EE2), a potent synthetic estrogen, was investigated in male and female Sprague-Dawley rats as part of a series of endocrine-active compounds, including genistein and nonylphenol. A method based on solid-phase extraction and LC with negative ion electrospray tandem mass spectrometric detection was developed and validated. The limit of detection in untreated rat serum was below 0.01 ng/ml (0.03 nM), the limit of quantification was 0.03 ng/ml (0.10 nM), the intra- and inter-day precision was 2-9%, and the intra- and inter-day accuracy was 89-94%. This method was used to determine the serum pharmacokinetics of EE2 in rats following oral gavage administration of 1 mg/kg body weight. EE2 was present in serum primarily in the unconjugated form at concentrations below 0.5 ng/ml. The maximal serum concentration was proportional to dose over the range of 0.04-0.5 mg/kg body weight and pharmacokinetic parameters were determined using model-independent analysis. Significant sex differences were observed for elimination half-times and volumes of distribution, but not for total serum clearance or maximal concentrations. The pharmacokinetic analysis of EE2 will be useful for comparing the toxicological effects of EE2 to those of other environmental estrogens in related rodent endocrine disruptor studies. PMID- 12906906 TI - Sensitive determination of tenofovir in human plasma samples using reversed-phase liquid chromatography. AB - A new high-performance liquid chromatography assay was developed for the determination of tenofovir, a nucleotide analogue, in plasma. A solid-liquid extraction procedure was coupled with a reversed-phase HPLC system. The system requires a mobile phase containing Na(2)HPO(4) buffer, tetrabutylammonium hydrogen sulfate and acetonitrile for different elution through a C(18) column with UV detection. The method proved to be accurate, precise and linear between 10 and 4000 ng/ml. The method was applied to determine trough levels of tenofovir in 11 HIV-infected patients with virologic failure under multiple antiretroviral therapy. This method was also successfully applied to a pharmacokinetic study in an HIV infected patient with renal failure. PMID- 12906907 TI - Novel purification system for 6xHis-tagged proteins by magnetic affinity separation. AB - We have developed a novel nickel-silica matrix for the generation of magnetic beads for metal-ion affinity chromatography. In contrast to magnetic Ni-NTA agarose beads, the novel particle type (SiMAC) consists of a magnetic core and a nickel-silica composite matrix with the nickel ions tightly integrated in the silica. This results in a much higher number of chelating groups compared with Ni NTA agarose beads. With the SiMAC beads, greatly improved purification of histidine-tagged proteins from crude bacterial extracts was achieved. The yield was at least twice as high as with conventional materials, the method is faster, since the coupling step is omitted and there is no need for handling toxic Ni(2+) salts. PMID- 12906908 TI - Metabolism and toxicological detection of the new designer drug 4'-methoxy-alpha pyrrolidinopropiophenone studied in rat urine using gas chromatography-mass spectrometry. AB - R,S-4'-Methoxy-alpha-pyrrolidinopropiophenone (MOPPP) is a new designer drug with assumed amphetamine-like effects, which has appeared on the illicit drug market. The aim of this study was to identify the MOPPP metabolites using solid-phase extraction, ethylation or acetylation as well as to develop a toxicological detection procedure in urine using solid-phase extraction, trimethylsilylation and GC-MS. Analysis of urine samples of rats treated with MOPPP revealed that MOPPP [limit of detection (S/N 3) was 100 ng/ml] was completely metabolized by demethylation of the methoxy group, hydroxylation of the pyrrolidine ring with subsequent dehydrogenation to the corresponding lactam and/or oxidative desamination to the corresponding diketo compounds. To some extent, the demethylated MOPPP metabolites were hydroxylated with partial subsequent methylation in position 3'. The hydroxy groups were found to be partly conjugated. Based on these data, MOPPP could be detected in urine via its metabolites by full-scan GC-MS using MS for screening and library search for identification by comparison of the spectra with reference spectra. PMID- 12906909 TI - Enantiomeric separation of TAPP, H-Tyr-(D)Ala-Phe-Phe-NH(2), by capillary electrophoresis using 18-crown-6-tetracarboxylic acid as a chiral selector. AB - A capillary electrophoresis method for the enantiomeric separation of the tetrapeptide H-Tyr-(D)Ala-Phe-Phe-NH(2) (TAPP), has been developed and validated. The preferred background electrolyte (BGE) consisted of 0.1 M aqueous phosphoric acid adjusted to pH 3.0 with triethanolamine. The chiral selectors 18-crown-6 tetracarboxylic acid (18C6H(4)) and heptakis(2,6-di-O-methyl)-beta-cyclodextrin (2,6-DM-beta-CD) were compared and the crown ether 18C6H(4) was found to be superior. The separation of TAPP and its enantiomer was accomplished within 30 min with a resolution greater than 3.5. The method was then investigated with respect to selectivity, linearity, accuracy, range, precision, detection limit (DL), quantitation limit (QL) and robustness, essentially following International Conference of Harmonisation (ICH) guidelines for the validation of analytical methods. The DL and QL for the TAPP enantiomer were found to be 0.3 and 0.8%, respectively, at the target TAPP concentration of 1 mg/ml. Robustness was tested using a full factorial design for the following four experimental variables varied at two levels: pH of the BGE, chiral selector concentration in the BGE, phosphoric acid concentration in the BGE, and temperature. The method showed good performance with respect to all of the validation parameters, and proved to be robust to changes in the experimental parameters within the tested domain. PMID- 12906910 TI - Extraction and quantitation of carfentanil and naltrexone in goat plasma with liquid chromatography-mass spectrometry. AB - This method is the first analytical method for the detection and quantitation of carfentanil and naltrexone at clinically relevant concentrations using liquid chromatography-mass spectrometry. Samples were alkalinized with 100 microl of 1 M NaOH and extracted 2x with 2 ml of toluene. The extractions were combined and dried under N(2) at 40 degrees C in a H(2)O bath. Chromatography was performed using a Zirchrom PBD column and a mobile phase of 30:70 acetonitrile/10 mM ammonium acetate and 0.1 mM citrate (pH=4.4) at a flow rate of 0.3 ml/min. The lower limit of quantitation was 8.5 pg/ml for carfentanil and 0.21 ng/ml for naltrexone. PMID- 12906911 TI - Simple and sensitive high-performance liquid chromatography method for simultaneous determination of urinary free cortisol and 6beta-hydroxycortisol in routine practice. For CYP 3A4 activity evaluation in basal conditions and after grapefruit juice intake. AB - Cytochrome p450 3A4 activity displays a wide variability. The urinary 6beta hydroxycortisol to cortisol ratio, as a non-invasive assay, can be useful for its pretherapeutic characterization. We developed an HPLC-UV method preceded by liquid-liquid extraction for assessment of this ratio in clinical practice. Urine was collected on second void morning-spot sample. Percentage recoveries were high and reproducible. The 6beta-hydroxycortisol to cortisol ratio ranged from 1.6 to 9.9 in 12 Caucasian healthy volunteers. It was reduced by 30 to 70% after ingestion of white grapefruit juice, a CYP3A4 inhibitor. Our method, simple, sensitive and accurate, could be helpful for determination of CYP 3A4 activity before oral chemotherapy, or for the monitoring of the use of grapefruit juice as a pharmacological modulator. PMID- 12906912 TI - Validated method for the determination of six metabolites derived from artichoke leaf extract in human plasma by high-performance liquid chromatography coulometric-array detection. AB - A validated method was developed for the simultaneous determination of the hydroxycinnamates caffeic (CA), dihydrocaffeic (DHCA), ferulic (FA), dihydroferulic (DHFA), and isoferulic acid (IFA) and the flavonoid luteolin (LUT) in human plasma as metabolites derived from artichoke leaf extract. The method involves sample preparation followed by separation using high-performance liquid chromatography on reversed-phase material with a polar endcapping (Aqua-C(18), 250 x 4.6 mm). Selectivity and sensitivity towards the target compounds were achieved by electrochemical array detection (CoulArray). Calibration curves were constructed in the ranges 2.1-51.7 ng x mL(-1) (CA), 2.0-76.7 ng mL(-1) (DHCA), 2.2-53.7 ng x mL(-1) (FA), 2.1-79.2 ng x mL(-1) (DHFA), 1.1-52.6 ng x mL(-1) (IFA) and 2.1-258.6 ng x mL(-1) (LUT). Linearity could be shown for all target compounds over the entire calibration range. Values for within-day and between day precision and accuracy were in accordance with the international guidelines for validation of bioanalytical methods. It is concluded that this newly developed method is appropriate for analysing samples from bioavailability and pharmacokinetic studies after oral administration of artichoke leaf extract. PMID- 12906913 TI - Metabolism and toxicological detection of the new designer drug 3',4' methylenedioxy-alpha-pyrrolidinopropiophenone studied in urine using gas chromatography-mass spectrometry. AB - R,S-3',4'-Methylenedioxy-alpha-pyrrolidinopropiophenone (MDPPP) is a new designer drug with assumed amphetamine-like effects, which has appeared on the illicit drug market. The aim of this study was to identify the MDPPP metabolites using solid-phase extraction, ethylation or acetylation as well as to develop a toxicological detection procedure in urine using solid-phase extraction, trimethylsilylation and GC-MS. Analysis of urine samples of rats treated with MDPPP revealed that MDPPP was completely metabolized by demethylenation of the methylenedioxy group followed by partial 3'-methylation of the resulting catechol, oxidative desamination to the corresponding diketo compounds and/or hydroxylation of the pyrrolidine ring with subsequent dehydrogenation to the corresponding lactam. The hydroxy groups were found to be partly conjugated. Based on these data, MDPPP could be detected in urine via its metabolites by full scan GC-MS using mass chromatography for screening and library search for identification by comparison of the spectra with reference spectra. PMID- 12906914 TI - Simultaneous enantioselective separation of azelastine and three of its metabolites for the investigation of the enantiomeric metabolism in rats. I. Liquid chromatography-ionspray tandem mass spectrometry and electrokinetic capillary chromatography. AB - Enantioselective separation methods and the enantioselective determination of the anti-allergic drug azelastine and of three of its main phase I metabolites in a biological matrix underwent chromatographic and electrophoretic investigations. An enantioselective assay of a coupling of HPLC using a beta-cyclodextrin chiral stationary phase to ionspray tandem mass spectrometry is presented. Additionally, this assay is compared to another enantioselective assay using electrokinetic capillary chromatography with beta-cyclodextrin and carboxymethyl-beta cyclodextrin in polyacrylamide-coated capillaries. For capillary electrophoresis (CE) the importance of polyacrylamide coating for the validation of this separation method is highlighted. Extracted rat plasma samples of enantioselective metabolism studies were measured by both validated assays. Differences in the pharmacokinetics and pharmacodynamics were evaluated for the main substance azelastine and its main metabolite demethylazelastine. So, a first hint about the enantioselectivity of biotransformation of azelastine in rats was seen after oral application of either enantiomer or the racemate to rats. PMID- 12906915 TI - Relative mRNA expression of the lactate dehydrogenase A and B subunits as determined by simultaneous amplification and single strand conformation polymorphism. Relation with subunit enzyme activity. AB - To explore if it is correlated in human tumor cells that the expression of LDH homologous gene and LDH isoenzymes, we used RT-PCR-SSCP technique to measure the relative expression of genes with homologous sequences. The combination of PCR using common primers designed in the highly conserved regions and single-strand conformation polymorphism analysis of the products is used for quantitative determination of the proportions of LDH-A mRNA in human cancer cell lines. The proportion is compared with that of the activities of isoenzymes. The results indicated that the enzyme activity of LDH-A was consistent with mRNA levels in the human tumor cell. The present procedure using a single pair of primers for two fragments can overcome disadvantages in quantitative analysis using multiplex PCR. Template concentrations and PCR cycles did not affect the proportions of LDH A and LDH-B in the product. PMID- 12906916 TI - Specific method for determination of OSI-774 and its metabolite OSI-420 in human plasma by using liquid chromatography-tandem mass spectrometry. AB - A new simple and specific method was developed and validated for the quantitative determination of OSI-774 (Tarceva, Erlotinib) and its metabolite, OSI-420, in human plasma. Sample pretreatment involved a single protein precipitation step with acetonitrile. The analytes were separated on Waters X-Terra C(18) (50 x 2.1 mm I.D., 3.5 microm) analytical column and eluted with acetonitrile-water mobile (70:30, v/v) containing 0.1% formic acid. The analytes of interest were monitored by tandem mass spectrometry with electrospray positive ionization. The overall extraction efficiency was greater than 88% for OSI-774 and 62% for OSI-420, with values for within-day and between-day precision and accuracy of <15%. Compared to previous assays, this method is simple, specific, and reproducible and will be used to characterize the plasma pharmacokinetics of OSI-774 at doses of 50 to 150 mg to optimize treatment with this agent. PMID- 12906917 TI - Determination of plasma testosterone using a simple liquid chromatographic method. AB - A simple and sensitive high-performance liquid chromatographic (HPLC) method using ultraviolet detection was developed for the determination of testosterone in human plasma. Testosterone and the internal standard, griseofulvin, were extracted from 0.50 ml plasma sample using a mixture of dichloromethane-2,2,4 trimethylpentane (3:2, v/v). The mobile phase, consisted of 0.02 M sodium dihydrogenphosphate-acetonitrile-methanol (51:47:2, v/v) adjusted to pH 3.1 and delivered to a C(18) analytical column (150 x 4.6 mm I.D., 4 microm particles) at a flow-rate of 1 ml/min while the detection wavelength was set at 240 nm with a sensitivity range of 0.005 a.u.f.s. The method has a quantification limit of 1.6 ng/ml. Recoveries of testosterone were all greater than 92% over the linear concentration range of 1.6-400 ng/ml while that of griseofulvin was approximately 95%. The within- and between-day RSD values were all less than 8% while the accuracy values ranged from 96.0 to 106.0% over the concentration range studied. The method was applied to the analysis of early morning plasma testosterone levels of 12 healthy human male volunteers. The levels were found to range from 3.1 to 8.4 ng/ml, within the normal range reported in the literature. PMID- 12906918 TI - On the significance of the role of cellular stress response reactions in the toxic actions of dioxin. AB - Dioxin is known to cause many toxic effects that vary greatly in different tissues, ages, genders, and species. In this review, an attempt has been made to sort out major signaling pathways involved in the expression of the toxicities of dioxin. The major strategy adopted in analyzing its major signaling pathways is to view the toxic actions of dioxin as the result of the Ah receptor-mediated expression of a major cellular emergency stress response signal. Evidence pointing to the similarities between the symptoms of poisoning by dioxin and those produced by chronic administration of typical stressors, particularly lipopolysaccharides (LPS), bacterial endotoxins, has been assembled and analyzed. The common symptoms are wasting syndrome, atherosclerosis, fatty liver, and thymic atrophy. On the other hand, oxidative stress caused by cytochrome P450 induction is one of the typical stresses of dioxin poisoning, but not LPS poisoning. One of the major means through which dioxin triggers stress responses via "stress-activated kinase pathways" is stimulation of the cellular production of cytokines/autocrines, particularly growth factors. In the case of hepatocytes for instance, transforming growth factor-alpha plays a pivotal role in the dioxin induced activation of the epidermal growth factor receptor and the extracellular signal-related kinase pathway, which acts as a signal to suppress apoptosis induced by cellular stress. These observations as well as additional experimental data support the idea that one of the major functions of the Ah receptor could be the elicitation of cellular stress response reactions. Another key point in understanding the toxic action of dioxin is that, unlike other cases of stressors, dioxin signaling becomes chronically sustained because of its extreme persistence in the human body, its half-life of 7-10 years, and its selective accumulation in fatty target tissues. PMID- 12906919 TI - Analysis of cell cycle regulation by 1-mono-O-acyl-3-O-(alpha-D-sulfoquinovosyl) glyceride (SQMG), an inhibitor of eukaryotic DNA polymerases. AB - One of the sulfo-lipids, 1-mono-O-acyl-3-O-(alpha-D-sulfoquinovosyl)-glyceride (SQMG), potently and selectively inhibited the activity of mammalian DNA polymerases. SQMG was also a potent apoptosis inducer and the SQMG effect occurred through the induction of G1 arrest with a reduction in the proportion of cells in the S phase. SQMG clearly increased the levels of p53 and p21 proteins, but did not induce the expression of p27 and p16 proteins. SQMG markedly reduced the pRb protein level and inhibited pRb phosphorylation after 48hr. These results suggested that SQMG activates the G1 checkpoint as a result of the DNA polymerase inhibition, and then promotes a p53-dependent apoptotic response. Since aphidicolin, a well-known replicative DNA polymerase inhibitor, did not promote these protein expressions, the apoptosis-inducing pathway by SQMG differs from that by aphidicolin. PMID- 12906920 TI - Characterization of sequence-dependent synergy between ZD1839 ("Iressa") and oxaliplatin. AB - ZD1839 ("Iressa"), a selective epidermal growth factor receptor tyrosine kinase inhibitor (EGFR-TKI), is currently undergoing preclinical and clinical evaluation in several solid tumors. The present study aimed to assess the effect of ZD1839 in combination with oxaliplatin in the colon cancer cell lines HT-29 and LoVo. For in vitro chemosensitivity testing, cells were treated with serial dilutions of each drug sequentially at a fixed ratio of doses that corresponded to 1/20, 1/10, 1/5, 1/2, 1, 1.5 and 2 times the individual IC(50) values. Oxaliplatin followed by ZD1839 produced a synergistic effect. In contrast, oxaliplatin following ZD1839 exhibited an additive effect at best. Mass spectrometry examination revealed that ZD1839 modestly enhanced cellular oxaliplatin accumulation and platinum-DNA (Pt-DNA) adducts (P>0.05). In additional studies, high-performance liquid chromatography revealed that oxaliplatin had no effect on ZD1839 accumulation. In contrast, ZD1839 markedly inhibited removal of Pt-DNA adducts (P<0.05). With oxaliplatin treatment (1 day) followed by ZD1839 (1 day), then incubation with drug-free medium (1 day), 90% of Pt-DNA adducts remained. Apoptosis examination revealed that oxaliplatin-induced apoptosis was prolonged by sequential oxaliplatin followed by ZD1839 treatment compared with oxaliplatin alone. Further experiments revealed that ZD1839 decreased cellular gamma glutamyltransferase activity. CONCLUSIONS: The above observations provide a mechanistic explanation for the synergy of oxaliplatin followed by ZD1839, and suggest that this treatment combination warrants further preclinical and clinical investigation. PMID- 12906921 TI - Hepatobiliary elimination of bile acid-modified oligodeoxynucleotides in Wistar and TR- rats: evidence for mrp2 as carrier for oligodeoxynucleotides. AB - As therapeutic antisense tools, oligonucleotides (ODNs) must enter cells to bind to their target structures. ODNs distribute in nearly each tissue with relatively high concentrations in kidney and liver from where excretion into urine and bile occurs. To investigate mechanisms involved in hepatic ODN transport, normal mixed backbone phosphodiester/phosphorothioate ODNs (n-ODN) and two different bile acid conjugated mixed backbone ODNs (1BA-ODN and 2BA-ODN) were applied to two different rat strains, normal Wistar rats and Wistar TR- rats. In normal Wistar rats, concentration-dependent hepatobiliary elimination of the ODNs was observed with a remarkable increase of excretion of the cholic acid BA-ODN conjugates. In contrast to normal Wistar rats, n-ODN excretion into bile by TR- rats, a mutant Wistar rat strain lacking a functional multidrug resistance-associated protein 2 (mrp2) at the canalicular membrane, was strongly diminished, whereas these rats excreted an ODN conjugated with two cholic acid molecules (2BA-ODN) into bile. Concomitant application of substrates transported by mrp2 such as bromosulfophthalein (BSP) or the synthetic chlorogenic acid derivative S 3025 significantly reduced the biliary appearance of normal ODN and 2BA-ODN in Wistar rats and also in TR- rats. To inhibit the expression of cRNA derived from the Na+ -dependent taurocholate cotransporting polypeptide (Ntcp), antisense ODNs were constructed which fully retained the antisense properties when coupled with two bile acid molecules. The results indicate that ODNs are secreted via the mrp2 into bile. In the absence of mrp2, further excretory transport systems with affinity for bile acids seem to be relevant for their excretion. The results further indicate that bile acid tagged ODNs are useful tools for liver specific antisense therapy. PMID- 12906922 TI - Influence of tumor necrosis factor-alpha on the expression and function of P glycoprotein in an immortalised rat brain capillary endothelial cell line, GPNT. AB - Drug cerebral pharmacokinetics may be altered in the case of inflammatory diseases. This may be due to a modification of drug transport through the blood brain barrier, in particular through drug interaction with the membrane efflux transporter, P-glycoprotein. The objective of this study was to investigate the influence of the inflammatory cytokine, tumor necrosis factor (TNF)-alpha, on the functionality and expression of P-glycoprotein, and on mdr1a and mdr1b mRNA expression in immortalised rat brain endothelial cells, GPNT. Cells were treated with TNF-alpha for 4 days. Levels of mdr1a and mdr1b mRNAs were quantitated using real-time RT-PCR analysis and expression of P-glycoprotein was analyzed by Western blot. The functionality of P-glycoprotein was studied by following the accumulation of [3H]vinblastine in the cells without and with a pre-treatment with a P-glycoprotein inhibitor, GF120918. TNF-alpha increased the levels of mdr1a and mdr1b mRNAs while no effect was observed on protein expression. TNF alpha increased [3H]vinblastine accumulation indicating a time and concentration dependent decrease of P-glycoprotein activity. This effect was eliminated when the cells were pre-treated with GF120918. Our observation of a decrease in P glycoprotein activity could suggest that in the case of inflammatory diseases, brain delivery of P-glycoprotein-dependent drugs can be enhanced. PMID- 12906923 TI - A role for increased mRNA stability in the induction of endothelin-1 synthesis by lipopolysaccharide. AB - An association exists between infection and cardiovascular diseases, including atherosclerosis, stroke and myocardial infarction. This may involve endothelin-1 (ET-1) which has been implicated in these and other vascular pathologies. ET-1 synthesis is controlled primarily by the level of its mRNA and numerous stimuli, including infection, lead to elevated ET-1 levels. Here, we have investigated the regulation of ET-1 release and preproET-1 (ppET-1) mRNA in bovine aortic endothelial cells by lipopolysaccharide (LPS). ET-1 release from bovine aortic endothelial cells was stimulated by LPS and reporter gene assays implicated LPS induced ppET-1 transcription. However, changes in transcription were modest compared to increases in ET-1 synthesis. Therefore, ppET-1 mRNA levels were measured by real-time reverse transcription-polymerase chain reaction. The effect of LPS on ppET-1 mRNA levels was more marked than on transcription (1.2-fold increase in transcription vs. 5.5-fold increase in ppET-1 mRNA). Analysis of ppET 1 mRNA stability by real-time reverse transcription-polymerase chain reaction showed that LPS increased its half-life by approximately 2-fold. Thus, upregulated ppET-1 mRNA and hence increased ET-1 synthesis may be due to both increased transcription and reduced mRNA degradation. These effects of LPS on mRNA stability may be a key mechanism in vascular pathologies through which many proteins are induced in response to infection. PMID- 12906924 TI - Different mechanisms for gamma-glutamyltransferase-dependent resistance to carboplatin and cisplatin. AB - In this work, we investigated the effect of gamma-glutamyltransferase (GGT) overexpression on cell viability after carboplatin treatment and compared with cisplatin. Carboplatin challenge of HeLa cells induced GGT and glutamate-cystine ligase (GCL) activities by 2- and 1.4-fold, respectively and concomitantly increased the intracellular reduced glutathione (GSH) level (1.5-fold). To study the role of GGT, HeLa-GGT cells, a stably transfected cell line overexpressing GGT (120-150 mU/mg protein) and the parental HeLa cells (10-15 mU/mg protein) were used. Both cell lines exhibited comparable viability (IC(50) approximately 150 microM) after carboplatin treatment when cultured in standard (250 microM cystine) medium. Culture in low (50 microM) cystine medium resulted in a dramatic decrease (approximately 90%) of the intracellular GSH level and to a 2.5-fold increase of carboplatin cytotoxicity (IC(50) approximately 60 microM). When GSH (50 microM) was included in the culture medium, only HeLa-GGT cells exhibited increased resistance to carboplatin. Using partially purified GGT from HeLa-GGT cells, we show that cisplatin forms adducts with cysteinylglycine, depending only on GGT activity whereas carboplatin did not efficiently react with cysteinylglycine. Thus, in this model system, GGT activity can affect platinum drugs cytotoxocity by two different ways: cisplatin can be detoxified extracellularly after reaction with the -SH group of cysteinylglycine; in the case of carboplatin, the supply of GSH precursors, initiated by GGT, increases the intracellular level of the tripeptide and provides enhanced defensive mechanisms to the cell. PMID- 12906925 TI - The study of the quercetin action on human erythrocyte membranes. AB - Quercetin is a naturally occurring flavonoid that exerts multiple pharmacological effects. In our previous study, we showed that quercetin greatly affects the lipid membrane. In this report, a study of quercetin on human erythrocyte membrane has been performed to determine the influence of this flavonoid on the fluidity and the conformational changes of membrane proteins. An additional aim of the study was to find how quercetin presence affects the resistance of membrane to haemolytic agents. The results showed that incorporation of quercetin into the erythrocyte membranes caused the changes of the partition coefficient of the Tempo spin label between the water and polar head group phases. In the studies, the W/S ratio has been used as a monitor of changes in protein conformation and in the environment within the membrane. It was observed that quercetin caused an increase in protein-protein interactions in human erythrocyte membranes. Haemolytic action of quercetin in the dark was also investigated. This compound showed protective effect against hypotonic haemolysis. However, in the heat-induced haemolysis quercetin caused acceleration of haemolysis. Dark reaction of erythrocyte with quercetin resulted in a shrinkage of the cells and alteration of their shapes. From the results we have concluded that modification of erythrocyte membrane by quercetin proceeds via reaction with membrane lipids and proteins. PMID- 12906926 TI - Identification of gene expression profiles predicting tumor cell response to L alanosine. AB - The methylthioadenosine phosphorylase (MTAP) gene gained considerable interest as therapeutic target for tumors with the 9p21 deletion. This gene maps to 9p21 and loss of this chromosomal region in tumors offers an unique opportunity for chemoselective treatment, since MTAP is an important salvage enzyme for the formation of adenine that is needed for DNA synthesis. L-Alanosine, an antibiotic from Streptomyces alanosinicus, blocks the common de novo purine biosynthesis pathway and, thereby, inhibits tumor cells with MTAP deficiency. Normal cells escape the detrimental effects of L-alanosine due to their proficiency in the MTAP salvage pathway. The present analysis was undertaken to gain insights into the molecular architecture of tumor cells that determines the response to L alanosine apart from the MTAP gene. Analysis of cell doubling times and IC(50) values for L-alanosine showed that slowly growing cell lines were more resistant to L-alanosine than rapidly growing ones. Mining the database of the National Cancer Institute (N.C.I.), for the mRNA expression of 9706 genes in 60 cell lines by means of Kendall's tau-test, false discovery rate calculation, and hierarchical cluster analysis pointed to 11 genes or expressed sequence tags whose mRNA expression correlated with the IC(50) values for L-alanosine. Furthermore, we tested L-alanosine for cross-resistance in multidrug-resistant cell lines which overexpress selectively either the P-glycoprotein/MDR1 (CEM/ADR5000), MRP1 (HL-60/AR), or BCRP (MDA-MB-231-BCRP) genes. None of the multidrug-resistant cell lines was cross-resistant to L-alanosine indicating that L-alanosine may be suitable to treat multidrug-resistant, refractory tumors in the clinic. Finally, the IC(50) values for L-alanosine of the 60 cell lines were correlated to the p53 mutational status and expression of p53 downstream genes. We found that p53 mutated cell lines were more resistant to L-alanosine than p53 wild type cell lines. PMID- 12906927 TI - A dual mechanism of action of the anticancer agent F 11782 on human topoisomerase II alpha. AB - F 11782 is a novel epipodophyllotoxin that targets eukaryotic topoisomerases and inhibits enzyme binding to DNA. While F 11782 has not been found to stabilize either topoisomerase I or topoisomerase II covalent complexes, drug treatment appears to result in DNA damage. F 11782 has also been shown to inhibit the DNA nucleotide excision repair (NER) pathway. Bisdioxopiperazine-resistant small cell lung cancer (SCLC) OC-NYH/Y165S and Chinese hamster ovary (CHO) CHO/159-1 cells having functional Y49F and Y165S mutations in the topoisomerase II alpha isoform were both resistant to F 11782. The catalytic activity of purified human Y50F and Y165S mutant topoisomerase II alpha (Y50F in the human protein corresponds to Y49F in the CHO protein) was likewise resistant to the inhibitory action of F 11782. F 11782 was also found to induce a non-covalent salt-stable complex of human topoisomerase II with DNA that was ATP-independent. F 11782 thus displays a dual mechanism of action on human topoisomerase II alpha, reducing its affinity for DNA while also stabilizing the protein bound in the form of a salt-stable complex. Our results suggest that topoisomerase II alpha is a target of F 11782 in vivo, and that F 11782 may act as a novel topoisomerase II poison. PMID- 12906928 TI - Regulation of pancreatic stellate cell function in vitro: biological and molecular effects of all-trans retinoic acid. AB - Pancreatic stellate cells (PSCs) are essentially involved in the development of pancreatic fibrosis, a constant feature of chronic pancreatitis and pancreatic cancer. Profibrogenic mediators, such as ethanol metabolites and cytokines, induce a PSC activation process that involves proliferation, enhanced production of extracellular matrix proteins and a phenotypic transition towards myofibroblasts which includes a loss of the characteristic retinoid-containing fat droplets. Here, we have analysed how exogenous all-trans retinoic acid (ATRA) affects activation of rat PSCs induced by sustained culture. Bromodeoxyuridine incorporation assays indicated an ATRA-dependent inhibition of DNA synthesis. In contrast, ATRA did not affect expression of alpha-smooth muscle actin, a protein typical for myofibroblasts. Quantification of [3H]proline incorporation revealed a diminished collagen production in ATRA-treated PSCs. Furthermore, zymography experiments showed that supernatants of ATRA-exposed PSC cultures contained higher levels of matrix metalloproteinase-9 but not of matrix metalloproteinase-2 than untreated controls. At the level of intracellular signalling, ATRA had no effect on extracellular signal-regulated kinase activation after incubation of PSCs with the mitogen platelet-derived growth factor (PDGF). In addition, PDGF induced DNA binding of activator protein-1 (AP-1) transcription factors was not inhibited by ATRA treatment. Luciferase reporter gene assays, however, revealed an ATRA-dependent transrepression of AP-1 in PDGF-stimulated PSCs. Together, the results indicate that exogenous ATRA displays inhibitory effects on PSC proliferation and collagen synthesis but does not block phenotypic transition towards myofibroblasts. We hypothesise that inhibition of AP-1 signalling may be involved in the mediation of biological effects of ATRA on PSCs. PMID- 12906929 TI - Inhibition of prostaglandin D2 clearance in rat hepatocytes by the thromboxane receptor antagonists daltroban and ifetroban and the thromboxane synthase inhibitor furegrelate. AB - Prostanoids, i.e. prostaglandins and thromboxane, regulate liver-specific functions both in homeostasis and during defense reactions. For example, prostanoids are released from Kupffer cells, the resident liver macrophages, in response to the inflammatory mediator anaphylatoxin C5a, and mediate an enhanced glucose output from hepatocytes as energy supply. In perfused rat livers, the thromboxane receptor antagonist daltroban enhanced C5a-induced prostanoid overflow and reduced glucose output. It was the aim of this study to elucidate whether daltroban interfered with prostanoid release from Kupffer cells or prostanoid clearance by hepatocytes, and/or whether it directly influenced prostanoid-dependent glucose metabolism in these cells. In perfused rat livers, daltroban enhanced prostaglandin (PG)D(2) overflow not only after infusion of C5a (15-fold), but also after PGD(2) (10-fold). Neither daltroban nor another receptor antagonist, ifetroban, or the thromboxane synthase inhibitor furegrelate enhanced prostanoid release from Kupffer cells. In contrast, all inhibitors reduced clearance, i.e. uptake and degradation, of PGD(2) by hepatocytes: within 5 min uptake of 1 nmol/L PGD(2) was reduced from 43+/-5 fmol (controls) to 22+/-6 fmol (daltroban), 24+/-6 fmol (ifetroban) and 21+/-6 fmol (furegrelate). PGD(2) in the medium was reduced to 39+/-7% in the controls, but remained at 93+/-9%, 93+/-11% and 60+/-3% in the presence of the inhibitors. PGD(2)-dependent glucose output in the perfused liver or activation of glycogen phosphorylase in isolated hepatocytes remained unaffected by daltroban. These data clearly demonstrate that the thromboxane-inhibitors reduced PGD(2) clearance by hepatocytes, presumably by inhibition of prostanoid transport into the cells. In contrast, they did not interfere with PGD(2)-dependent glucose metabolism, suggesting an independent mechanism for the inhibition of glucose output from the liver. PMID- 12906930 TI - Degradation of endomorphin-2 at the supraspinal level in mice is initiated by dipeptidyl peptidase IV: an in vitro and in vivo study. AB - Endomorphin-2 (Tyr-Pro-Phe-PheNH(2)) was discovered as an endogenous ligand for the mu-opioid receptor. The physiological function of endomorphin-2 as a neurotransmitter or neuromodulator may cease through the rapid enzymatic process in the synapse of brain, as for other neuropeptides. The present study was conducted to examine the metabolism of endomorphin-2 by synaptic membranes prepared from mouse brain. Major metabolites were free tyrosine, free phenylalanine, Tyr-Pro and PheNH(2). Both the degradation of endomorphin-2 and the accumulation of major metabolites were inhibited by specific inhibitors of dipeptidyl peptidase IV, such as diprotin A and B. On the other hand, the accumulation of Phe-PheNH(2) and Pro-Phe-PheNH(2) was increased in the presence of bestatin, an aminopeptidase inhibitor, whereas that of free phenylalanine and PheNH(2) was decreased. Furthermore, purified dipeptidyl peptidase IV hydrolyzed endomorphin-2 at the cleavage site, Pro(2)-Phe(3) bond. Thus, degradation of endomorphin-2 by brain synaptic membranes seems to take place mainly through the cleavage of Pro(2)-Phe(3) bond by dipeptidyl peptidase IV, followed by release of free phenylalanine and PheNH(2) from the liberated fragment, Phe-PheNH(2) by aminopeptidase. We have also examined that the effect of diprotin A on the antinociception induced by intracerebroventricularly administered endomorphin-2 in the mouse paw withdrawal test. Diprotin A simultaneously injected with endomorphin-2 enhanced endomorphin-2-induced antinociception. These results indicate that dipeptidyl peptidase IV may be an important peptidase responsible for terminating endomorphin-2-induced antinociception at the supraspinal level in mice. PMID- 12906931 TI - Antidiabetic and antimalarial biguanide drugs are metal-interactive antiproteolytic agents. AB - Various biguanide derivatives are used as antihyperglycemic and antimalarial drugs (e.g., 1,1-dimethyl biguanide (metformin), phenylethyl biguanide (phenformin), N-(4-chlorophenyl)-N'-(isopropyl)-imidodicarbonimidic diamide (proguanil)); however, no common mechanism has been suggested in these controversial therapeutic actions. Biguanides bind endogenous metals that inhibit cysteine proteases independently, e.g., Zn(2+), Cu(2+), Fe(3+). Here, various biguanide derivatives are reported to be metal-interactive inhibitors of cathepsin B from mammals and falcipain-2 from Plasmodium falciparum. Structural homologies were identified among the Phe-Arg protease substrate motif and the metal complexes of phenformin and proguanil. Molecular modeling revealed that the position of the scissile amide substrate bond corresponds to the biguanide complexed inhibitory metal when the phenyl groups are homologously aligned. Binding of the phenformin-metal complex within the active site of human cathepsin B was modeled with computational docking. A major binding mode involved binding of the drug phenyl group at the protease S2 subsite, and the complexed inhibitory metal shared between the drug and the protease Cys29-His199 catalytic pair. Cysteine protease inhibition was assayed with carbobenzyloxy-PHE-ARG-7 aminomethylcoumarin substrate. In the absence of metal ions, phenformin was a weakly competitive protease inhibitor (apparent K(i) several microM); however, metformin was noninhibitory. In contrast, the metal complexes of both metformin and phenformin were protease inhibitors with potency at therapeutic concentrations. Biguanide-metal complexes were more potent cysteine protease inhibitors than either the biguanide or metal ions alone, i.e., synergistic. Similar to chloroquine, therapeutic extracellular concentrations of metformin, phenformin, and proguanil caused metal-interactive inhibition of lysosomal protein degradation as bioassayed in primary tissue using perfused myocardium. The biguanide moiety is identified as a past and future structural scaffold for synthesis of many protease inhibitors. Results are discussed in relation to Zn(2+)-interactive inhibition of insulin degradation in hormone target tissues, and Fe(3+)-interactive inhibition of hemoglobin degradation in parasite food vacuoles. Previous studies on insulin hypercatabolism and insulin resistance are speculatively reviewed in light of present findings. PMID- 12906932 TI - Down-regulation of c-jun N-terminal kinase-activator protein-1 signaling pathway by Ginkgo biloba extract in human peripheral blood T cells. AB - The activation of T lymphocytes contributes to inflammatory process of cardiovascular and cerebrovascular diseases. We investigated the effects of the extract of Ginkgo biloba (EGb), an ancient plant preserving antioxidant property, on phorbol 12-myristate 13-acetate+ionomycin or anti-CD3+anti-CD28 monoclonal antibodies-activated T cells. Human peripheral blood T cells were negatively selected from whole blood. Cytokines were measured by ELISA, cell surface markers by flow cytometry and the activities of transcription factors and kinases were determined by electrophoresis mobility shift assays, kinase assays and transfection assays. We showed that EGb inhibited several cytokines, including tumor necrosis factor-alpha, interleukin (IL)-2, IL-4 and interferon-gamma production from activated T cells. Electrophoresis mobility shift assay analysis indicated that EGb down-regulated activator protein-1 (AP-1) but not nuclear factor kappa B DNA-binding activity. In addition, EGb inhibited c-jun N-terminal kinase but not extracellular signal regulated protein kinase activity. The inhibitory specificity on AP-1 by EGb was also demonstrated in transfection assays. The inhibition of AP-1 signaling pathway in T cells by EGb provides a support for its efficacy in cardiovascular and cerebrovascular diseases and raises a therapeutic potential for this drug in activated T cell-mediated pathologies. PMID- 12906933 TI - Characterization of a plasma membrane-resident albumin-binding protein associated with the proliferation of estrogen-target, serum-sensitive cells. AB - Estrogens control the proliferation of their target cells through a receptor mediated pathway. Recently presented evidence suggests that estradiol cancels the proliferative inhibition exerted by human albumin (HA) and recombinant human albumin (rHA) on estrogen-target serum-sensitive cells (indirect-negative hypothesis). We postulate that this mechanism requires the presence of a plasma membrane estrogen receptor (mER) and a plasma membrane albumin-binding protein (mABP). Direct evidence confirming the presence of mERalpha in MCF7 cells has recently been presented. Herein, we now show that Western blot analysis of purified T47D membrane proteins with the C542 ERalpha specific monoclonal antibody also revealed specific, multiple M(r) mERs (67, 110, and 130k M(r)). In addition, Western blot analysis with an ABP antiserum revealed a potential 60k M(r) ABP in both MCF7 and T47D plasma membrane extracts. No such evidence was observed in similar extracts from ER-negative, serum-insensitive MDA-MB231 cells. Ligand blot analysis of similar plasma membrane extracts with bovine serum albumin confirmed the presence of a 60k M(r) ABP in MCF7 and T47D cells; again, no such evidence was observed in comparable extracts from MDA-MB231 cells. Fluorescence and confocal microscopy of MCF7 cells fixed in 2.0% paraformaldehyde/0.1% glutaraldehyde identified specific membrane ABP antigenic sites by immunocytochemistry. Serum-insensitive MDA-MB231 cells fixed and labeled similarly did not exhibit this mABP. These results suggest that the proposed mABP is expressed only in serum-sensitive estrogen-target cells and is not expressed in cells insensitive to the proliferative inhibition of HA and rHA. Also, the present data suggest that the proposed mABP may be the recognition mechanism by which both HA and rHA inhibit MCF7 and T47D cell proliferation. PMID- 12906934 TI - 27-Hydroxylation of 7- and 8-dehydrocholesterol in Smith-Lemli-Opitz syndrome: a novel metabolic pathway. AB - Smith-Lemli-Opitz syndrome (SLOS) is attributable to mutations in the gene coding for 7-dehydrocholesterol reductase. Low to absent enzyme activity accounts for the accumulation of both 7-dehydrocholesterol and 8-dehydrocholesterol in plasma and other tissues. Since oxysterols can participate in the regulation of cholesterol homeostasis, we examined the possibility that they are formed from these dehydrocholesterol intermediates. In patients with SLOS, we found serum levels of 27-hydroxy-7-dehydrocholesterol ranging from 0.1 to 0.25micro M and evidence for circulating levels of 27-hydroxy-8-dehydrocholesterol (0.04-0.51 micro M). Picomolar quantities of 27-hydroxy-7-dehydrocholesterol were identified in normal individuals. Biologic activities of 27-hydroxy-7-dehydrocholesterol were found to include inhibition of sterol synthesis and the activation of nuclear receptor LXRalpha but not that of LXRbeta. These activities occurred at concentrations found in plasma and presumably at those existing in tissues. Thus, patients with SLOS have increased levels of metabolites derived from intermediates in cholesterol synthesis that are biologically active and may contribute to the regulation of cholesterol synthesis in vivo. PMID- 12906935 TI - Probing the active site of aromatase with 2-methyl-substituted androstenedione analogs. AB - To gain insight into the spatial nature of the androstenedione (AD) binding (active) site of aromatase in relation to the catalytic function of the enzyme, we synthesized 2,2-dimethylAD (4), 2beta- and 2alpha-methylADs (5 and 6), 19 oxygenated derivatives of compounds 4 and 6, and 2-methyleneAD (17), and we then tested their inhibitory activity as well as their aromatase reaction (aromatization for 2-methyl and 2-methylene analogs or 19-oxygenation for 2,2 dimethyl steroids) with human placental aromatase. 2-Methyl and 2-methylene steroids 5, 6, and 17 were good competitive inhibitors of aromatase (K(i)=22 68nM), but less effective compared to the 2,2-dimethyl analog 4 (K(i)=8.8nM), indicating that a combination of 2beta- and 2alpha-methyl moieties is essential for the formation of a thermodynamically stable inhibitor-aromatase complex. A series of 2alpha-methyl steroids were good substrates for aromatase, whereas 2beta-methyl steroid 5 was an extremely poor substrate, and a series of 2,2 dimethyl steroids did not serve as substrate, suggesting that a 2beta-methyl moiety of the 2,2-dimethyl and 2beta-methyl steroids would prevent the aromatase reaction probably due to steric hindrance in each case. The 2-methylene compound 17 was also aromatized to produce 2-methylestrogen with a low conversion rate where the 1,4-diene structure may have been created before the C(10)-C(19) bond cleavage. Kinetic analysis of the aromatization of androgens revealed that a good substrate was not essentially a good inhibitor for aromatase. PMID- 12906936 TI - HPLC and ELISA analyses of larval bile acids from Pacific and western brook lampreys. AB - Comparative studies were performed on two native lamprey species, Pacific lamprey (Lampetra tridentata) and western brook lamprey (Lampetra richardsoni) from the Pacific coast along with sea lamprey (Petromyzon marinus) from the Great Lakes, to investigate their bile acid production and release. HPLC and ELISA analyses of the gall bladders and liver extract revealed that the major bile acid compound from Pacific and western brook larval lampreys was petromyzonol sulfate (PZS), previously identified as a migratory pheromone in larval sea lamprey. An ELISA for PZS has been developed in a working range of 20 pg-10 ng per well. The tissue concentrations of PZS in gall bladder were 127.40, 145.86, and 276.96 micro g/g body mass in sea lamprey, Pacific lamprey, and western brook lamprey, respectively. Releasing rates for PZS in the three species were measured using ELISA to find that western brook and sea lamprey released PZS 20 times higher than Pacific lamprey did. Further studies are required to determine whether PZS is a chemical cue in Pacific and western brook lampreys. PMID- 12906937 TI - Differential effects of short-term prednisolone treatment on peripheral and abdominal subcutaneous thickness in children assessed by ultrasound. AB - Long-term glucocorticoid excess decreases peripheral and increases abdominal subcutaneous thickness. Short-term prednisolone treatment is used in the treatment of many acute and chronic conditions in children. The aim of the present study was to elucidate if changes in thickness of cutis, subcutis, or dermal water content may be induced by short-term prednisolone treatment in children. Twenty children with asthma aged 7.7-13.8 years were included in a double-blind, randomized, placebo-controlled crossover trial. Active treatment was 5mg prednisolone daily. Treatment, run-in, and wash-out periods were 1 week. On days 1 and 7 of each treatment period, 20 MHz ultrasound scanning of the skin was performed on the thigh, forearm, and abdomen. Prednisolone treatment was associated with decreases in the total thickness of the cutis and subcutis in the thigh (0.28 mm) and forearm (0.15 mm), and an increase in the abdomen (0.23 mm). During placebo treatment the thickness was increased in the thigh (0.07 mm) and abdomen (0.05 mm), and reduced in the forearm (0.03 mm). The differences between prednisolone and placebo treatment were statistically significant in the thigh (P=0.04). The increase in thickness in the abdomen during prednisolone treatment was statistically significantly different from the reductions in the thigh (P=0.03) and forearm (P=0.05). There were no statistically significant differences in the dermal thickness or water content during prednisolone treatment compared to placebo.Short-term treatment with 5mg prednisolone daily may cause differential effects in peripheral and abdominal subcutaneous thickness in children. PMID- 12906938 TI - Structure-activity relationships of 3-deoxy androgens as aromatase inhibitors. Synthesis and biochemical studies of 4-substituted 4-ene and 5-ene steroids. AB - As part of our investigation into the structure-activity relationship of a novel class of aromatase inhibitors, two series of 3-deoxy androgens, androst-5-en-17 ones with a non-polar alkoxy (5 and 6), alkyl (20-22), or phenylalkyl (23 and 24) group at C-4beta and 4-acyloxyandrost-4-en-17-ones (29-32, and 34) were synthesized and evaluated. The 4beta-alkyl and 4beta-phenylalkyl compounds were obtained through reaction of 4alpha,5alpha-epoxy steroid (8) with RMgBr (R: alkyl and phenylalkyl) followed by dehydration of the 4beta-substituted 5alpha-hydroxy products (15-19) with SOCl(2) as key reactions. Acylation of 4alpha,5alpha-diol (25) with (RCO)(2)O in pyridine and subsequent dehydration with SOCl(2) gave the 4-acyloxy steroids. All of the steroids studied, except for 4-acetoxy-19-ol (34) that was a non-competitive inhibitor of human placental aromatase, blocked aromatase activity in a competitive manner. 4-Benzoyloxy- and 4-acetoxy steroids (31) and (32) were the most powerful inhibitors of aromatase (K(i)=70 and 60nM, respectively). Elongation of an acetoxy group in a series of 4-acyloxy steroids or a methyl group in a series of 4beta-alkyl steroids decreased affinity for aromatase principally in relation to carbon number of the acyl or alkyl function. The present findings are potentially useful for understanding the spatial and electronic nature of the binding site of aromatase as well as for developing effective aromatase inhibitors. PMID- 12906939 TI - Anti-glucocorticoid effects of progesterone in vivo on rat adipose tissue metabolism. AB - Steroid hormones seem to be important for adipose tissue metabolism and accumulation. As progesterone has been suggested to modulate the glucocorticoid effects, the interactions between glucocortioid and progesterone on adipose tissue metabolism were investigated.Forty-eight male Wistar rats were adrenectomized and divided into four groups; controls (treated with vehicle only), dexamethasone treated (10 micro g per rat), progesterone treated (5mg per rat) and the last group received both dexamethasone and progesterone. The dexamethasone-treated group had a significant loss of body weight and smaller intra-abdominal fat depots compared to the control group in addition, dexamethasone increased LPL-activity and increased catecholamine stimulated lipolysis. When progesterone was given concomitantly the dexamethasone effects on adipose tissue mass, LPL-activity and lipolysis were blocked. When given alone progesterone had no influence on body weight, amount of adipose tissue, lipolysis or LPL-activity. These data indicate that progesterone acts as an anti glucocorticoid in adipose tissue in vivo, thus attenuating the glucocorticoid effect on adipose tissue metabolism. PMID- 12906940 TI - Cortisol secretion in the elderly. Influence of age, sex and cardiovascular disease in a Chinese population. AB - Adrenal function and aging have been the object of intense interest in recent years. In this study we analyzed morning (08:00 h) serum cortisol concentrations from a sample of Chinese subjects aged from 31 to 110 years. These levels differed according to age, health status and sex, although the sex difference was confirmed only among the healthy elderly. These results suggest that age (older than 60 years), disease and male sex are associated with increased morning serum cortisol levels in a Chinese population. PMID- 12906941 TI - Robust, automatic spike sorting using mixtures of multivariate t-distributions. AB - A number of recent methods developed for automatic classification of multiunit neural activity rely on a Gaussian model of the variability of individual waveforms and the statistical methods of Gaussian mixture decomposition. Recent evidence has shown that the Gaussian model does not accurately capture the multivariate statistics of the waveform samples' distribution. We present further data demonstrating non-Gaussian statistics, and show that the multivariate t distribution, a wide-tailed family of distributions, provides a significantly better fit to the true statistics. We introduce an adaptation of a new expectation-maximization based competitive mixture decomposition algorithm and show that it efficiently and reliably performs mixture decomposition of t distributions. Our algorithm determines the number of units in multiunit neural recordings, even in the presence of significant noise contamination resulting from random threshold crossings and overlapping spikes. PMID- 12906942 TI - Quantifying location-specific information in the discharge of rat hippocampal place cells. AB - The informational content in the location-specific discharge of rat hippocampal cells is usually quantified by an average for the entire behaviorally accessible space. In contrast to such "global" information measures, we consider here information that can be obtained from "local" spike counts at each position. The properties of these local information measures are first illustrated using simulated data with predetermined distributions of location-specific spike counts. Next, place cell recordings from rats foraging in a cylindrical arena with two cue cards on its walls are analyzed; time windows as short as 100 ms were used to accumulate spike counts in locations. We show that information at the centers of firing fields is higher for fields nearer to the cues. Neither firing rates or "global" information measures detected differences between fields near and far from the cues. Thus, analyses of the location-specific information provides a new valuable tool for studying the location-specific activity of rat hippocampal cells. Generalizations of location-specific information can be used to investigate place cell responses to other factors such as running speed or the state of the hippocampal EEG in addition to current position. PMID- 12906943 TI - Surface visualization of electromagnetic brain activity. AB - Advances in hardware and software have made possible the reconstruction of brain activity from non-invasive electrophysiological measurements over a large part of the brain. The appreciation of the information content in the data is enhanced when relevant anatomical detail is also available for visualization. Different neuroscientific questions give rise to different requirements for optimal superposition of structure and function. Most available software deal with scalar measures of activity, especially hemodynamic changes. In contrast, the electrophysiological observables are generated by electrical activity, which depends on the synchrony of neuronal assemblies and the geometry of the local cortical surface. We describe methods for segmentation and visualization of spatio-temporal brain activity, which allow the interplay of geometry and scalar as well as vector properties of the current density directly in the representations. The utility of these methods is demonstrated through displays of tomographic reconstructions of early sensory processing in the somatosensory and visual modality extracted from magnetoencephalography (MEG) data. The activation course characteristic to a specific area could be observed as current density or statistical maps independently and/or contrasted to the activity in other areas or the whole brain. MEG and functional magnetic resonance imaging (fMRI) activations were simultaneously visualized. Integrating and visualizing complementary functional data into a single environment helps evaluating analysis and understanding structure/function relationships in normal and diseased brain. PMID- 12906944 TI - A simple method for immunocytochemical staining with multiple rabbit polyclonal antibodies. AB - We report a single step, simple, repeatable, rapid and reliable technique for simultaneous immunocytochemical staining with two or more rabbit polyclonal antibodies. This technique, which we have dubbed the "Pretty Poly" method, is based on conjugating the antibodies with commercially available, fluorophore tagged Staphylococcal protein-A (SP-A). Staining is illustrated at the calyx type presynaptic nerve terminal of the chick ciliary ganglion with antibodies directed against three nerve terminal proteins: neurofilaments of the axonal cytoskeleton, and two secretory vesicle proteins, SV2 and cysteine string protein (CSP). Images were deblurred with an iterative deconvolution protocol. Staining with a single polyclonal antibody was bright and had a resolution approaching light microscope limit. Treatment with two different polyclonal antibodies conjugated with contrasting dye-tagged protein-A resulted in double staining without significant crossover that was fully equivalent to the standard primary/secondary technique. The same single step protocol was used to stain with all three rabbit polyclonal antibodies or to combine the technique with a standard monoclonal primary/secondary antibody stain. Thus, the Pretty Poly protocol is a highly flexible, simple and yet effective staining technique that essentially solves the problem of co-staining with multiple polyclonal rabbit antibodies. PMID- 12906945 TI - Pixel-based criteria-oriented analysis of time-lapse Ca2+-fluorescence images. AB - Since its inception, the analysis of time-lapse video-images acquired during Ca2+ imaging experiments using fluorescence microscopy has been progressively optimized for achieving a high temporal resolution. In contrast, the spatial resolution of the acquired images is often compromised during analysis to varying degrees by the need to draw regions of interest (ROI). We developed a strategy to analyze images at the acquired spatial resolution-pixel-by-pixel, grouping all pixels based on criteria of interest (COI) in regard to their associated fluorescence values over time and visualizing the distributions of the pixel groups detected in a pseudo-colored map. We applied this pixel-based COI-strategy to the analysis of relative intracellular free calcium levels (Ca(i)(2+)) in attached cultured embryonic hippocampal cells under baseline and experimental conditions designed to evaluate the contribution of extracellular Ca2+ (Ca(e)(2+)) to baseline Ca(i)(2+) levels. We discovered distinct groups of Ca(e)(2+)-dependent Ca(i)(2+) regulation patterns emergent during the earliest phases of hippocampal cell differentiation, which were not limited to inter-cell differences. Thus, pixel-based COI-analysis of time-lapse images can be used to disclose distinct patterns of Ca(e)(2+)-dependent Ca(i)(2+) levels and their corresponding subcellular distributions in developing hippocampal cells. Such a strategy should be useful in studying the emergence and distribution of Ca(i)(2+) signaling at subcellular levels of resolution using fluorescence microscopy. PMID- 12906946 TI - Single neuron activity in the Drosophila larval CNS detected with calcium indicators. AB - Although the Drosophila larva has been extensively used for genetic studies of synaptic transmission and locomotion, neurophysiological studies have lagged because it is difficult to investigate circuitry and synaptic function in the larval central nervous system (CNS). Here we introduce an optical technique to monitor neuronal activity in the intact Drosophila larval CNS. We loaded neurons retrogradely through cut axons with dextran-conjugated calcium indicators. Fluorescence responses to changes in the concentration of intracellular calcium are sufficiently fast and large to monitor electrical activity in single neurons. Responses to action potentials were detected in motor neuron cell bodies, axons, neurites, dendrites and sensory neuron afferents identified by genetically targeted green fluorescent protein expression. Our findings provide an experimental procedure for testing synaptic function and connectivity within the intact larval CNS. PMID- 12906947 TI - Polyethylenimine improves the transfection efficiency of primary cultures of post mitotic rat fetal hypothalamic neurons. AB - Analysis of gene regulatory sequences in primary cultures of neurons has been hampered by inefficient transfection of post-mitotic neurons with reporter plasmids. We describe detailed conditions that allowed a significant improvement of transfection efficiency in primary cultures of serum-supplemented rat fetal hypothalamic cells. Transfected cells expressed the green fluorescent protein (GFP) under the control of the strong but non-cell-specific cytomegalo virus (CMV) promoter or under the thyrotropin-releasing hormone (TRH) promoter, to direct expression only in TRH neurons. Using the CMV promoter-GFP plasmid, we tested several commercially available transfection reagents; the best results were obtained with polyethylenimine (PEI) and Lipofectamine 2000. We optimized the transfection procedure with PEI because it rendered more reproducible results. Transfection with PEI was optimal when cells were transfected at a cellular density of 2.9 x 10(6) cells in 35-mm dishes, with 10 microg of DNA, a PEI/DNA ratio of 8.8 and PEI pH of 6.9. Using these conditions, we were able to detect GFP positive neurons after transfecting the TRH promoter-GFP plasmid. GFP positive cells were successfully purified by FACS. This opens the possibility to use transfection of mammalian CNS post-mitotic neurons for new applications including the purification of specific neuronal subtypes. PMID- 12906948 TI - Increased cortical excitability induced by transcranial DC and peripheral nerve stimulation. AB - This study investigated the effect of short periods of simultaneous weak anodal direct current (DC) stimulation and peripheral ulnar nerve (ES) stimulation on corticospinal excitability. The experiments involved repeated testing of ten normal subjects with four different protocols: (1) No Stimulation; (2) DC only; (3) ES only; (4) DC plus ES. Motor evoked potentials (MEP) were recorded from the First Dorsal Interosseous (FDI); Abductor Digiti Minimi (ADM) and Flexor Carpi Ulnaris (FCU). The baseline MEP amplitude was compared with that obtained immediately after the first set of stimulation, after the second set of stimulation, 15 min post stimulation and 30 min after stimulation. No significant change was seen with the No Stimulation and ES Only protocols. FDI showed a significant change in the DC protocol after the second set of stimulation but the changes were not present 15 or 30 min after. The DC plus ES protocol showed FDI changes that were significant after the second set of stimulation with the elevations persisting when tested 15 and 30 min post intervention. These observations show that a period of anodal DC stimulation preceding a period of ulnar nerve stimulation resulted in significant and persistent elevations in cortical excitability. PMID- 12906949 TI - Retrograde transport of sodium selenite and intracellular injection of micro ruby: a combined method to describe the morphology of zinc-rich neurones. AB - Zinc is found in synaptic vesicles in a large number of glutamatergic systems. Its involvement in neurotransmission and neurological disorders has been suggested. There are methods for tracing these circuits, but they do not fill the dendritic tree. In this study, extracellular selenite injections in vivo were combined with intracellular injection of fluorochromes in fixed tissue to reveal the morphology of these zinc-rich neurones. Intraperitoneal and intracerebral injections of sodium selenite alone or intracerebral injections of selenite combined with bisbenzimide were made in the visual cortex of the rat in order to locate the somata of zinc-rich neurones. After 24 h of retrograde transport, animals were killed and fluorescent markers were injected intracellularly into fixed slices to show neuronal morphology: (a) Lucifer Yellow (LY) followed by biocytin, (b) LY coupled to biocytin or (c) micro-ruby (MR) (dextranamines bound to rhodamine and biotin). Double-labelled somata (selenite+fluorochrome) were plotted. Details of the dendritic morphology were then revealed by incubation in avidin-biotin complex and development in 3,3'-diaminobenzidine and H(2)O(2). Camera lucida drawings showed that zinc-rich neurones in layers II-III involved in cortico-cortical visual projections were typical pyramidal neurones. This technique is noteworthy for its analysis of the morphology (and connections) of zinc-rich neurones. PMID- 12906950 TI - Parenteral methadone: an essential medication for the treatment of pain. PMID- 12906951 TI - Choreiform movements after a single dose of methadone. PMID- 12906952 TI - Naloxone in treating central adverse effects during opioid titration for cancer pain. PMID- 12906953 TI - Poor social conditions, criminality and urban violence: Unmentioned barriers for effective cancer pain control at the end of life. PMID- 12906954 TI - The impact of demographic and disease-specific variables on pain in cancer patients. AB - The aim of this study was to examine to what extent demographic and disease specific variables affected pain in cancer patients. Two to three weeks after their last hospitalization, 1,453 cancer patients completed questionnaires measuring demographic variables, quality of life, and pain (EORTC-QLQ C-30). Response rate was 72.1%. Data on type of cancer and the severity of the disease were also compiled. Sixty percent of the sample reported some pain. Type of cancer, presence of metastases, and time until death were significant predictors of experienced pain. The patients' experience of pain was mainly associated with disease-specific variables. Sex, age, level of education, and co-habitation were not related to pain, but employment status was. The patients on disability pensions had significantly more pain than the patients who were working or studying. Special attention should be given to patients with advanced prostate cancer with a short time to live, as they reported the most pain. PMID- 12906955 TI - My mind is as clear as it used to be: A pilot study illustrating the difficulties of employing a single-item subjective screen to detect cognitive impairment in outpatients with cancer. AB - Oncology patients often complain that their "mind does not seem to be clear." This subjective perception, sometimes referred to as "chemo brain," may be due to situational stressors, psychological disorders, organic factors, or effects of neurotoxic medications. Cognitive decline cannot only diminish quality of life, but can also interfere with a patient's ability to make decisions regarding complex treatment issues. The current study investigated the utility of using item 11 of the Zung Self-Rating Depression Screen (ZSDS) as a cognitive screen. A sample of 61 ambulatory cancer patients completed this study. Participants were recruited from four sites of Community Cancer Care, Inc., in Indiana. A battery of cognitive instruments and psychosocial inventories was administered in a standardized order. The sample had a mean age of 58.6 years and comprised 57.4% (n=35) women and 42.6% (n=26) men. Item 11 of the ZSDS was not significantly correlated to the cognitive measures. Correlates of the perception of cognitive impairment were the Dementia Rating Scale (DRS) Attention Scale (r=-0.26, P<0.05) and the ZSDS total score (r=-0.29, P<0.05). Patients' perceptions of having a cognitive impairment determined by item 11 of the ZSDS was predicted by total score on the ZSDS (F=42.5, P<0.001), age (F=26.0, P<0.001), and score on the Stroop test (F=19.8, P<0.001). Analysis of sensitivity and specificity indicated that the single-item screen used in this study is not an accurate means for identifying oncology patients with actual cognitive impairment. We conclude that while the perception of cognitive impairment is common in cancer patients, there may be problems in interpreting the nature of these complaints, particularly in separating them from depressive preoccupation. PMID- 12906956 TI - Pain control in ambulatory cancer patients--can we do better? AB - To evaluate the degree of pain control among ambulatory cancer patients visiting the outpatient clinics of three oncology centers in south Israel, these patients were interviewed using the Brief Pain Inventory translated into Hebrew (BPI-Heb). Patients suffering from pain at least three times a week or reporting taking daily analgesics during the last two weeks were enrolled. Non-Hebrew speakers and patients too frail or ill were excluded. The study population included 218 subjects. Substantial pain was experienced by 77%, the majority was not adequately treated (81%), and 75% were undermedicated. The daily living activities of the majority of patients (64%) were moderately to severely impacted. Pain control was not associated with any of the sociodemographic or previous treatment profile variables, or by physicians' pain assessment. The physicians' and the patients' ratings of the extent to which pain interfered with the patients' activities fully agreed (+/-2) in fewer than half of the patients. Physicians estimated more severe pain levels, but underestimated its impact on everyday life. These data indicate that better pain control for ambulatory cancer patients is needed and that more information about patients' pain and its impact should be solicited. Further training of care providers is needed to improve the relief from cancer pain and the quality of life of patients. PMID- 12906957 TI - Use of oral and transdermal opioids among patients with metastatic cancer during the last year of life. AB - This study documents the use of oral and transdermal opioids among patients with metastatic cancer during their final year of life. Using a large, integrated health-insurance claims database, we identified all patients who had metastatic lung, breast, colorectal, prostate, or breast cancer and who also died in 1998 or 1999. We then examined all pharmacy claims for these patients over their final 12 months of life. A total of 2,132 patients were identified who met study entrance criteria. Among patients with bone metastases (n=717), 86.9% received opioids at some point during their final year of life; 71.2% of those without bone metastases (n=1,415) received them. Corresponding figures for long-acting opioids were 52.9% and 23.5%. Coverage ratios (total days supplied/total noninstitutionalized days) for any opioids and long-acting opioids were 25.1% and 12.5%, respectively, among patients with bone metastases, and 13.9% and 4.2% for those without bone metastases. During the final month of life, these ratios were 50.8% and 31.3%, and 28.7% and 13.1%. These relatively low rates of opioid use among patients with metastatic cancer in their final year of life suggest that pain in many cases may be suboptimally treated. PMID- 12906958 TI - The efficacy of acupressure and acustimulation wrist bands for the relief of chemotherapy-induced nausea and vomiting. A University of Rochester Cancer Center Community Clinical Oncology Program multicenter study. AB - As an adjunct to standard antiemetics for the relief of chemotherapy-induced nausea and vomiting (NV), 739 patients were randomly assigned to either: 1) acupressure bands, 2) an acustimulation band, or 3) a no band control condition. Patients in the acupressure condition experienced less nausea on the day of treatment compared to controls (P<0.05). There were no significant differences in delayed nausea or vomiting among the three treatment conditions. Additional analyses revealed pronounced gender differences. Men in the acustimulation condition, but not the acupressure condition, had less NV compared to controls (P<0.05). No significant differences among the three treatment conditions were observed in women, although the reduction in nausea on the day of treatment in the acupressure, compared to the no band condition, closely approached statistical significance (P=0.052). Expected efficacy of the bands was related to outcomes for the acupressure but not the acustimulation conditions. PMID- 12906959 TI - A pharmacokinetic study to compare two simultaneous 400 microg doses with a single 800 microg dose of oral transmucosal fentanyl citrate. AB - It is unknown whether two smaller doses of oral transmucosal fentanyl citrate (OTFC) administered simultaneously are pharmacokinetically equivalent to an identical dose administered as a single unit. This issue has important practical implications when patients are attempting to identify the appropriate dosage of OTFC to control their pain. This open-label, randomized, crossover design study compared the pharmacokinetics of two simultaneously consumed 400 microg OTFC doses with one 800 microg OTFC dose in 12 healthy volunteers. The two treatments were pharmacokinetically equivalent. The maximum concentration produced for each dosage group (Cmax) was 1.09 ng/ml for two 400 microg dose and 1.10 ng/ml for one 800 microg dose. Area under the curve (AUC) was 8.2 ng/ml.hr (SE=1.1) and 7.2 ng/ml.hr (SE=1.0). There were no significant differences between the treatment groups in either the time to peak concentration (Tmax) or the mean residence time (MRT). The results demonstrate the bioequivalence of two 400 microg with one 800 microg OTFC units. PMID- 12906960 TI - Pediatric cancer pain management practices and attitudes in China. AB - Children with cancer often receive less adequate pain management than adults. This survey was designed to examine current practices, knowledge, and attitudes pertaining to pediatric pain management in China. Based on the choice of opioid use and self-identification as having a conservative or liberal approach to pain management, we identified two clusters. Members of cluster 1 (60%) had more conservative attitudes and did not perceive that young children experienced pain. Members of cluster 2 (40%) had more liberal attitudes. Four factors predicted conservative membership: never having prescribed opioids for cancer pain (OR=2.67, CI=1.11-6.45), use of placebos for pain management (OR=2.32, CI=1.05 5.08), belief that the adequacy of pain management was fair or good (OR=2.10, CI=1.31-3.37), and belief that children are less sensitive to pain (OR=1.96, CI=1.02-3.76). The top barriers to optimal pediatric pain management were identified as fear of opioid addiction and inadequate knowledge of pain management. PMID- 12906961 TI - Nurses' willingness and the predictors of willingness to provide palliative care in rural communities of taiwan. AB - The aims of this study are to identify the willingness of district nurses to provide palliative care in the rural communities in Taiwan and determine the predictors of this willingness. A questionnaire was sent to all 1,121 community nurses of the 174 government health stations assigned to all the rural areas of Taiwan. The overall response rate was 86.4%, with 940 valid questionnaires retrieved. The majority of respondents (93.0%) expressed a willingness to provide palliative care if they encountered a terminal cancer patient. However, they would limit their services to consultation and referral (91.6% and 87.3%), and were less likely to provide home visiting (51.0%) or bereavement support of family (38.4%). With regard to knowledge, the accurate answers to the philosophy/principles and clinical practice of palliative care were 88.5% and 43%, respectively. The results of stepwise logistic regression analysis showed that "palliative care knowledge," "the belief in external control by authority," and "the belief in external control by chance" were the most significant variables related to the willingness of district nurses to provide palliative care (OR=12.83, 95% CI=2.46-66.76, P<0.01; OR=2.11, 95% CI=1.32-3.35, P<0.01; OR=0.54, 95% CI=0.31-0.95, P<0.05, respectively). However, regarding the content of services, the willingness to provide home visiting was affected positively by the level of information resources, subjective norms, and the belief in external control by authority. This willingness was negatively affected by age of the patient. These data suggest that effective training courses focused on practical knowledge of palliative care for these district nurses, the incorporation of the palliative care into nursing education, and active health policy administration are critical for the community palliative care movement in Taiwan. PMID- 12906962 TI - Hyperalgesia: an emerging iatrogenic syndrome. AB - Clinical reports suggest that opioids, intended to abolish pain, can unexpectedly produce hyperalgesia. This paradoxical effect may be mechanistically related to tolerance induced by increasing doses of opioids. Two case reports illustrate a syndrome characterized by increasing pain pursued by escalating opioid doses, which results in a worsening of the clinical picture. Several experimental data may help explain the course of this challenging clinical condition. In escalating opioid doses rapidly, a risk of opioid-induced hyperalgesia should be recognized, as higher doses of opioids may stimulate rather than inhibit the central nervous system by different mechanisms. Alternative procedures should be taken into consideration to break this cycle, should it occur. More data are needed to detect this condition, as currently no diagnostic information on specific markers, clinical or biochemical, exists. PMID- 12906963 TI - Ventricular repolarization components on the electrocardiogram: cellular basis and clinical significance. AB - Ventricular repolarization components on the surface electrocardiogram (ECG) include J (Osborn) waves, ST-segments, and T- and U-waves, which dynamically change in morphology under various pathophysiologic conditions and play an important role in the development of ventricular arrhythmias. Our primary objective in this review is to identify the ionic and cellular basis for ventricular repolarization components on the body surface ECG under normal and pathologic conditions, including a discussion of their clinical significance. A specific attempt to combine typical clinical ECG tracings with transmembrane electrical recordings is made to illustrate their logical linkage. A transmural voltage gradient during initial ventricular repolarization, which results from the presence of a prominent transient outward K(+) current (I(to))-mediated action potential (AP) notch in the epicardium, but not endocardium, manifests as a J-wave on the ECG. The J-wave is associated with the early repolarization syndrome and Brugada syndrome. ST-segment elevation, as seen in Brugada syndrome and acute myocardial ischemia, cannot be fully explained by using the classic concept of an "injury current" that flows from injured to uninjured myocardium. Rather, ST-segment elevation may be largely secondary to a loss of the AP dome in the epicardium, but not endocardium. The T-wave is a symbol of transmural dispersion of repolarization. The R-on-T phenomenon (an extrasystole originating on the T-wave of a preceding ventricular beat) is probably due to transmural propagation of phase 2 re-entry or phase 2 early after depolarization that could potentially initiate polymorphic ventricular tachycardia or fibrillation. PMID- 12906964 TI - Waiting times, revascularization modality, and outcomes after acute myocardial infarction at hospitals with and without on-site revascularization facilities in Canada. AB - OBJECTIVES: This study was designed to determine whether admission to a Canadian hospital with on-site revascularization (invasive hospital) affected revascularization choice, timing, and outcome compared with community (non invasive) hospitals. BACKGROUND: Health care systems in Canada are characterized by relative restraint in diffusion of tertiary cardiovascular services, with capacity for revascularization procedures concentrated in large regional referral centers. METHODS: We used linked administrative data and a clinical registry to follow-up 15,166 Ontario patients who underwent revascularization within the year after their index acute myocardial infarction (MI). Outcomes included recurrent urgent cardiac hospitalization, hospital bed-days, and death within the same year after the index admission. We adjusted for age, gender, socioeconomic status, illness severity, attending physician specialty, and academic hospital affiliation. RESULTS: After adjusting for baseline factors, patients admitted to invasive hospitals were more likely to receive angioplasty than bypass surgery (adjusted odd ratio: 1.85; 95% confidence interval: 1.68 to 2.04, p < 0.001). The converse pattern was seen for patients admitted to community hospitals. Median revascularization waiting times were significantly shorter at invasive hospitals (12 vs. 48 days, p < 0.001). Patients admitted to invasive hospitals had fewer cardiac re-admissions (41.5 vs. 68.9 events per 100 patients, p < 0.001) before their first revascularization and consumed fewer hospital bed-days (379 vs. 517 per 100 patients, p < 0.001). There were no differences in outcomes beyond revascularization. CONCLUSIONS: Outcome advantages associated with timely post-MI revascularization highlight the importance of organizing revascularization referral networks and facilitating access to revascularization for patients with acute coronary syndromes admitted to community hospitals in Canada. PMID- 12906965 TI - The early days after ST-segment elevation acute myocardial infarction: reconsidering the delayed invasive approach. PMID- 12906966 TI - The impact on coagulation of an intravenous loading dose in addition to a subcutaneous regimen of low-molecular-weight heparin in the initial treatment of acute coronary syndromes. AB - OBJECTIVES: We sought to quantify the impact of adding an intravenous loading dose to a subcutaneous regimen of enoxaparin in patients with an acute coronary syndrome (ACS). BACKGROUND: It is unclear whether an intravenous (i.v.) loading dose of enoxaparin should be added to a subcutaneous (s.c.) regimen in patients with ACS. METHODS: Patients admitted with ACS were randomized to i.v.+s.c.(n = 14) or s.c. alone (n = 11) enoxaparin treatment. Coagulation markers were measured at nine time points during the first 24 h of treatment. RESULTS: The i.v.+s.c. therapy immediately resulted in therapeutic anti-Xa levels, which remained significantly higher for 6 h compared with s.c. alone, without reaching excessively high levels. A rapid decrease of plasma prothrombin fragments 1+2 (F(1+2)) levels was observed as soon as 5 min after the i.v. injection (33% lower; p = 0.007), and these levels remained lower up to 2 h after the start of treatment compared with SQ alone. The ex vivo thrombin generation time was maximally prolonged at 5 min post-injection in the i.v.+s.c. group and remained significantly prolonged up to 6 h post-injection compared with s.c. alone. The tissue factor pathway inhibitor plasma activity was immediately increased by 194% with i.v.+s.c., whereas the maximum increase with s.c. alone was 47% at 3 h. CONCLUSIONS: Therapeutic plasma levels of enoxaparin are achieved significantly earlier by an i.v.+s.c. regimen compared with s.c. alone, without leading to unacceptably high levels. As the risk of thrombotic complications is greatest early after admission, the observed differences in antithrombotic effects may translate into a clinical benefit. However, this remains to be established. PMID- 12906967 TI - Percutaneous coronary interventions in octogenarians. glycoprotein IIb/IIIa receptor inhibitors' safety profile. AB - OBJECTIVES: This study was designed to evaluate the safety profile of glycoprotein IIb/IIIa receptor inhibitors (GPI) in octogenarians undergoing percutaneous coronary intervention (PCI). BACKGROUND: Patients > or =80 years old constitute the fastest growing segment of the U.S. population and have a high prevalence of coronary artery disease. Few data exist regarding the use of GPI during PCI in octogenarians, as these patients have been excluded from randomized clinical trials of GPI. METHODS: Consecutive patients > or =80 years old undergoing PCI between January 1998 and June 2001 were evaluated for clinical outcomes and bleeding complications. RESULTS: One thousand three hundred and ninety two of 14,308 patients (9.7%) undergoing PCI were > or =80 years old. Of these, 459 of 1,392 (33%) of the patients were treated with GPI. Octogenarians treated with GPI were more likely to present with acute coronary syndrome or infarction, receive stents, require an intra-aortic balloon pump, or undergo multi-vessel PCI. Glycoprotein receptor inhibitor use was associated with a higher rate of bleeding, but the transfusion rate was similar to that in patients who did not receive GPI (9.8% vs. 8.6%, p = NS). No cases of intracranial hemorrhage were observed. By multivariate analysis, GPI treatment was associated with longer hospitalization but did not independently predict the need for transfusion or affect mortality. CONCLUSIONS: Octogenarians have a high incidence of bleeding and need for transfusion after PCI. Although the use of GPI was associated with more access and non-access site bleeding and longer hospital stay, GPI treatment does not significantly increase the risk of transfusion or intracranial hemorrhage in this non-randomized cohort. PMID- 12906968 TI - Percutaneous coronary intervention in octogenarians and the safety of glycoprotein IIb/IIIa inhibitors. PMID- 12906969 TI - Nonelectrocardiographic evidence that both ischemic preconditioning and adenosine preconditioning exist in humans. AB - OBJECTIVES: The objective of this study was to use electrocardiogram (ECG) independent parameters to determine whether preconditioning (PC) exists in humans during percutaneous transluminal coronary angioplasty (PTCA). BACKGROUND: Several studies suggest that both ischemia and adenosine induce PC in the human heart during PTCA. However, because almost all of these studies relied on ST-segment shifts as indicators of the severity of ischemia, their conclusions continue to be questioned, and the very existence of ischemic or adenosine PC in humans remains controversial. METHODS: Eighteen patients received either intracoronary adenosine (n = 9) or normal saline (n = 9); 10 min later, they underwent PTCA (three 2-min balloon inflations 5 min apart). RESULTS: Compared with the first inflation, in untreated patients the second and third inflations were associated with less systolic dysfunction (two-dimensional echocardiography), less diastolic dysfunction (color M-mode echocardiography), less lactate production, and less H(+) release into the great cardiac venous blood. In adenosine-treated patients, the extent of all of these abnormalities during the first inflation was less than in untreated patients and did not change with subsequent inflations. CONCLUSIONS: Previous exposure to a brief episode of ischemia (first balloon inflation) or to adenosine produces concordant decreases in ECG, subjective, mechanical, and metabolic manifestations of ischemia during PTCA. These data support the concept that both ischemic PC and pharmacologic PC exist in humans and that PTCA is a useful clinical setting in which to discern their mechanism. PMID- 12906970 TI - Patients with early diabetic heart disease demonstrate a normal myocardial response to dobutamine. AB - OBJECTIVES: We sought to use quantitative markers of the regional left ventricular (LV) response to stress to infer whether diabetic cardiomyopathy is associated with ischemia. BACKGROUND: Diabetic cardiomyopathy has been identified in clinical and experimental studies, but its cause remains unclear. METHODS: We studied 41 diabetic patients with normal resting LV function and a normal dobutamine echo and 41 control subjects with a low probability of coronary disease. Peak myocardial systolic velocity (Sm) and early diastolic velocity (Em) in each segment were averaged, and mean Sm and Em were compared between diabetic patients and controls and among different stages of dobutamine stress. RESULTS: Both Sm and Em progressively increased from rest to peak dobutamine stress. In the diabetic group, Sm was significantly lower than in control subjects at baseline (4.2 +/- 0.9 cm/s vs. 4.7 +/- 0.9 cm/s, p = 0.012). However, Sm at a low dose (6.0 +/- 1.3), before peak (8.4 +/- 1.8), and at peak stress (8.9 +/- 1.8) in diabetic patients was not significantly different from that of controls (6.3 +/- 1.4, 8.9 +/- 1.6, and 9.6 +/- 2.1 cm/s, respectively). The Em (cm/s) in the diabetic group (rest: 4.2 +/- 1.2; low dose: 5.0 +/- 1.4; pre-peak: 5.3 +/- 1.1; peak: 5.9 +/- 1.5) was significantly lower than that of controls (rest: 5.8 +/- 1.5; low dose: 6.6 +/- 1.5; pre-peak: 6.9 +/- 1.3; peak: 7.3 +/- 1.7; all p < 0.001). However, the absolute and relative increases in Sm or Em from rest to peak stress were similar in diabetic and control groups. CONCLUSIONS: Subtle LV dysfunction is present in diabetic patients without overt cardiac disease. The normal response to stress suggests that ischemia due to small-vessel disease may not be important in early diabetic heart muscle disease. PMID- 12906971 TI - Diabetic cardiomyopathy. the importance of being earliest. PMID- 12906972 TI - Echocardiographic prediction of left ventricular dysfunction after mitral valve repair for mitral regurgitation as an indicator to decide the optimal timing of repair. AB - OBJECTIVES: This study sought to determine whether echocardiography before mitral valve repair (MVR) for mitral regurgitation (MR) was predictive of postoperative left ventricular (LV) dysfunction and useful for deciding the optimal timing of repair. BACKGROUND: Some reports have shown that the preoperative echocardiographic data of left ventricular ejection fraction (LVEF) and left ventricular end-systolic diameter (LVDs) were good predictors of postoperative LV dysfunction. However, few reports were based on long-term follow-up data of large numbers of patients who underwent MVR in the last decade. METHODS: A total of 274 patients with moderate or severe MR underwent MVR between October 1, 1991, and September 30, 2000. Among them, 171 patients who had both an operation for isolated MR due to degenerative pathology and a postoperative echocardiogram were studied. Postoperative echocardiograms were performed 3.9 +/- 2.4 years after the operation. The LVEF decreased from 66 +/- 10% before surgery to 63 +/- 11% after surgery (p < 0.0001). On univariate analysis, preoperative LVEF and LVDs correlated with postoperative LVEF (r = 0.41 and r = -0.39, respectively). Overall, postoperative LV dysfunction (defined as LVEF <50%) was not frequent (12%). However, the incidence of postoperative LV dysfunction was high in patients with preoperative LVEF <55% (38%) or LVDs > or =40 mm (23%). CONCLUSIONS: In patients with MR, the echocardiographic data of LVEF and LVDs were good predictors of postoperative LV dysfunction. When a decrease in LVEF or an increase in LVDs is detected, MVR should be considered to preserve postoperative LV function. PMID- 12906973 TI - Unexpected, dismal left ventricular function after surgery for mitral regurgitation: there is just no excuse for it anymore. PMID- 12906974 TI - Detection of viruses in myocardial tissues by polymerase chain reaction. evidence of adenovirus as a common cause of myocarditis in children and adults. AB - OBJECTIVES: The purpose of this study was to analyze cardiac tissue and blood for viral genomes using polymerase chain reaction (PCR) to define the common viral etiologies of myocarditis by age group. BACKGROUND: Enteroviruses are considered the most common cause of myocarditis at all ages. Diagnosis relies on viral cultures, serology, and cardiac histology, which lack sensitivity, as well as PCR. However, in many cases enteroviruses are not detected. METHODS: Cardiac samples were obtained for PCR analysis from patients with myocarditis (n = 624) and dilated cardiomyopathy (DCM) (n = 149). Patients were analyzed by age group, including neonates (n = 116), infants (n = 191), toddlers (n = 87), children (n = 110), adolescents (n = 92), and adults (n = 177). After nucleic acids had been extracted from an endomyocardial biopsy, an explant, or autopsy samples, PCR and reverse transcription PCR were performed to detect the genomic sequences of enterovirus, adenovirus, cytomegalovirus (CMV), herpes simplex virus (HSV), Epstein-Barr virus (EBV), parvovirus, respiratory syncytial virus (RSV), and influenza A virus. RESULTS: Viral genome was amplified (adenovirus = 142, enterovirus = 85, CMV = 18, parvovirus = 6, influenza A = 5, HSV = 5, EBV = 3, RSV = 1) from 239 (38%) of the 624 samples from myocarditis patients, including 26 patient samples in which dual infection was found. Virus was detected in 30 (20%) of 149 DCM patient samples; only adenovirus (n = 18) and enterovirus (n = 12) were detected. CONCLUSIONS: Polymerase chain reaction identified adenovirus as the most common virus in the myocardium of children and adults with myocarditis and DCM. Although enteroviruses are also found in these patients, they appear to be a less common cause of myocarditis than adenovirus. PMID- 12906975 TI - Eradication of viral myocarditis: is there hope? PMID- 12906976 TI - Plasma C-reactive protein as a marker of cardiac allograft vasculopathy in heart transplant recipients. AB - OBJECTIVES: This study was initiated to determine whether heart transplant recipients (HTRs) with cardiac allograft vasculopathy (CAV) have increased levels of high-sensitivity C-reactive protein (hsCRP) and to examine whether an increase in hsCRP after heart transplantation predicts the development of CAV. Furthermore, the effect of pravastatin on plasma levels of hsCRP in HTRs was investigated. BACKGROUND: The relationship between CAV and hsCRP, as well as the effect of statins on hsCRP in HTRs, has not been well established. METHODS: On referral for their annual angiographic control study, 150 consecutive HTRs (mean 6.5 years since transplantation) were included. Plasma levels of hsCRP were measured before angiography and compared with patients with (n = 52) and without (n = 98) CAV. In 49 of these patients, we additionally analyzed hsCRP in blood samples stored from their six-month visit after the transplantation procedure. Furthermore, in a randomized, crossover study, hsCRP was analyzed in 17 male HTRs before and after six weeks of treatment with 20 mg pravastatin. RESULTS: Median levels of CRP were elevated among patients with CAV compared with those with normal angiograms [3.86 (1.78 to 7.00) vs. 1.08 (0.72 to 2.13) mg/l, p < 0.001]. Prospectively evaluated hsCRP levels from six months to follow-up were significantly higher among those who developed CAV compared with those with normal angiograms [+2.76 (1.56 to 5.00) vs. +0.07 (-0.57 to 0.41) mg/l, p < 0.001]. On multivariate analysis, the increase in hsCRP was the only significant predictor of CAV. Six weeks of treatment with pravastatin significantly reduced hsCRP levels by 25%, without any relation to changes in lipid values. CONCLUSIONS: Elevated plasma levels of CRP are associated with angiographic evidence of CAV, and the increase in hsCRP is a strong predictor of development of CAV. Statin treatment reduces levels of hsCRP and should be used in HTRs, regardless of their lipid levels. PMID- 12906977 TI - C-Reactive protein and cardiac allograft vasculopathy. is inflammation the critical link? PMID- 12906978 TI - Cardiac resynchronization therapy can reverse abnormal myocardial strain distribution in patients with heart failure and left bundle branch block. AB - OBJECTIVES: We studied the effects of cardiac resynchronization therapy (CRT) on regional myocardial strain distribution, as determined by echocardiographic strain rate (SR) imaging. BACKGROUND: Dilated hearts with left bundle branch block (LBBB) have an abnormal redistribution of myocardial fiber strain. The effects of CRT on such abnormal strain patterns are unknown. METHODS: We studied 18 patients (12 males and 6 females; mean age 65 +/- 11 years [range 33 to 76 years]) with symptomatic systolic heart failure and LBBB. Doppler myocardial imaging studies were performed to acquire regional longitudinal systolic velocity (cm/s), systolic SR (s(-1)), and systolic strain (%) data from the basal and mid segments of the septum and lateral wall before and after CRT. By convention, negative SR and strain values indicate longitudinal shortening. RESULTS: Before CRT, mid-septal peak SR and peak strain were lower than in the mid-lateral wall (peak SR: -0.79 +/- 0.5 [septum] vs. -1.35 +/- 0.8 [lateral wall], p < 0.05; peak strain: -7 +/- 5 [septum] vs. -11 +/- 5 [lateral wall], p < 0.05). This relationship was reversed during CRT (peak SR: -1.35 +/- 0.8 [septum] vs. -0.93 +/- 0.6 [lateral wall], p < 0.05; peak strain: -11 +/- 6 [septum] vs. -7 +/- 6 [lateral wall], p < 0.05). Cardiac resynchronization therapy reversed the septal lateral difference in mid-segmental peak strain from -46 +/- 94 ms (LBBB) to 17 +/- 92 ms (CRT; p < 0.05). CONCLUSIONS: Left bundle branch block can lead to a significant redistribution of abnormal myocardial fiber strains. These abnormal changes in the extent and timing of septal-lateral strain relationships can be reversed by CRT. The noninvasive identification of specific abnormal but reversible strain patterns should help to improve patient selection for CRT. PMID- 12906979 TI - Cost implications of testing strategy in patients with syncope: randomized assessment of syncope trial. AB - OBJECTIVES: We sought to assess the cost implications of two investigation strategies in patients with unexplained syncope. BACKGROUND: Establishing a diagnosis in patients with unexplained syncope is complicated by infrequent and unpredictable events. The cost-effectiveness of immediate, prolonged monitoring as an alternative to conventional diagnostic strategies has not been studied. METHODS: Sixty patients (age 66 +/- 14 years; 33 males) with unexplained syncope and LV ejection fraction >35% were randomized to conventional testing with an external loop recorder, tilt and electrophysiologic (EP) testing, or prolonged monitoring with an implantable loop recorder with one-year monitoring. If patients remained undiagnosed after their assigned strategy, they were offered a crossover to the alternate strategy. Cost analysis of the two testing strategies was performed. RESULTS: Fourteen of 30 patients who were being monitored were diagnosed at a cost of 2,731 Canadian dollars +/- 285 Canadian dollars per patient and 5,852 Canadian dollars +/- 610 Canadian dollars per diagnosis. In contrast, only six of 30 conventional patients were diagnosed (20% vs. 47%, p = 0.029), at a cost of 1,683 Canadian dollars +/- 505 Canadian dollars per patient (p < 0.0001) and 8,414 Canadian dollars +/- 2,527 Canadian dollars per diagnosis (p < 0.0001). After crossover, a diagnosis was obtained in 1 of 5 patients undergoing conventional testing, compared with 8 of 21 patients who completed monitoring (20% vs. 38%, p = 0.44). Overall, a strategy of monitoring followed by tilt and EP testing was associated with a diagnostic yield of 50%, at a cost of 2,937 Canadian dollars +/- 579 Canadian dollars per patient and 5,875 Canadian dollars +/- 1,159 Canadian dollars per diagnosis. Conventional testing followed by monitoring was associated with a diagnostic yield of 47%, at a greater cost of 3,683 Canadian dollars +/- 1,490 Canadian dollars per patient (p = 0.013) and a greater cost per diagnosis (7,891 Canadian dollars +/- 3,193 Canadian dollars, p = 0.002). CONCLUSIONS: A strategy of primary monitoring is more cost-effective than conventional testing in establishing a diagnosis in recurrent unexplained syncope. PMID- 12906980 TI - Implantable loop recorders. dollars and sense. PMID- 12906981 TI - Relationship of contractile function to transmural extent of infarction in patients with chronic coronary artery disease. AB - OBJECTIVES: We sought to determine the relationship of contractile function to the transmural extent of infarction (TEI) in patients with chronic coronary artery disease. BACKGROUND: In the setting of reperfused, chronic myocardial infarction (MI), the relationship of contractile function to the TEI has not been established. METHODS: We studied function by cine magnetic resonance imaging (MRI) and the TEI by contrast-enhanced MRI in 31 patients with single-vessel disease 162 +/- 62 days after reperfused first MI. RESULTS: Of all 516 segments with MI, blinded observers were unable to detect abnormal thickening in 193 (37%), and wall thickening measured quantitatively in these segments was 66 +/- 28%. Of the 193 segments, 163 (84%) were infarcts limited to the subendocardium. The average TEI reached 53% before half of the patients had abnormal contractile function. When patients with small MI (< or =5% of total left ventricular [LV] mass) were excluded, the average TEI reached 43% before half the patients had abnormal function. In subjects with small MI (< or =5% of total LV mass [n = 13]), even segments with TEI >75% had normal function (14 of 14) because they were surrounded by normally moving neighbor segments. CONCLUSIONS: In the setting of reperfused chronic MI, the TEI approaches 50% before contractile dysfunction can be systematically identified. Contractile function cannot be used to rule out chronic MI. PMID- 12906982 TI - Cardiovascular magnetic resonance of acute myocardial infarction at a very early stage. AB - OBJECTIVES: Very early changes in myocardial tissue composition during acute myocardial infarction (AMI) are difficult to assess in vivo. Cardiovascular magnetic resonance (CMR) imaging provides techniques for visualizing tissue pathology. BACKGROUND: The diagnostic role of CMR in very acute stages of myocardial infarction is uncertain. We investigated signal intensity changes beginning within 60 min after acute coronary occlusion in patients undergoing therapeutic septal artery embolization. METHODS: We investigated eight patients with hypertrophic obstructive cardiomyopathy undergoing interventional septal artery embolization by applying microparticles to reduce left ventricular outflow tract obstruction. In a clinical 1.5-tesla (T) CMR system, we visualized infarct related myocardial signal by T(1)-weighted sequences before and 20 min after administration of contrast media (delayed enhancement) and edema-related signal by T(2)-weighted spin-echo sequences before and 58 +/- 14 min after the intervention as well as on days 1, 3, 7, 14, 28, 90, and 180 during follow-up. RESULTS: Infarct-related changes as defined by contrast enhancement were observed as early as 1 h after the intervention and during six months of follow-up. In contrast, infarct-related myocardial edema, as visualized by high signal intensity in T(2)-weighted spin-echo sequences, was not consistently detectable 1 h after acute arterial occlusion; this was possible in all subsequent studies until day 28. CONCLUSIONS: Contrast-enhanced magnetic resonance imaging detected infarct-related signal changes as early as 1 h after AMI in humans, whereas the sensitivity of edema-related signal changes was not sufficient during this very early stage. PMID- 12906983 TI - Magnetic resonance to assess the aortic valve area in aortic stenosis: how does it compare to current diagnostic standards? AB - OBJECTIVES: The purpose of the present study was to evaluate whether magnetic resonance (MR) planimetry of the aortic valve area (AVA) may prove to be a reliable, non-invasive diagnostic tool in the assessment of aortic valve stenosis, and how the results compare with current diagnostic standards. BACKGROUND: Current standard techniques for assessing the severity of aortic stenosis include transthoracic and transesophageal echocardiography (TEE) as well as transvalvular pressure measurements during cardiac catheterization. METHODS: Forty consecutive patients underwent cardiac catheterization, TEE, and MR. The AVA was estimated by direct planimetry (MR, TEE) or calculated indirectly via the peak systolic transvalvular gradient (catheter). Pressure gradients from cardiac catheterization and Doppler echocardiography were also compared. RESULTS: By MR, the mean AVA(max) was 0.91 +/- 0.25 cm(2); by TEE, AVA(max) was 0.89 +/- 0.28 cm(2); and by catheter, the AVA was calculated as 0.64 +/- 0.26 cm(2). Mean absolute differences in AVA were 0.02 cm(2) for MR versus TEE, 0.27 cm(2) for MR versus catheter, and 0.25 cm(2) for TEE versus catheter. Correlations for AVA(max) were r = 0.96 between MR and TEE, r = 0.47 between TEE and catheter, and r = 0.44 between MR and catheter. The correlation between Doppler and catheter gradients was r = 0.71. CONCLUSIONS: Magnetic resonance planimetry of the AVA correlates well with TEE and less well with the catheter-derived AVA. Invasive and Doppler pressure correlated less well than those obtained from planimetric techniques. Magnetic resonance planimetry of the AVA may provide an accurate, non invasive, well-tolerated alternative to invasive techniques and transthoracic echocardiography in the assessment of aortic stenosis. PMID- 12906984 TI - The ACE/DD genotype is associated with the extent of exercise-induced left ventricular growth in endurance athletes. AB - OBJECTIVES: We studied the impact of the angiotensin-converting enzyme (ACE)/DD genotype on morphologic and functional cardiac changes in adult endurance athletes. BACKGROUND: Trained athletes usually develop adaptive left ventricular hypertrophy (LVH), and ACE gene polymorphisms may regulate myocardial growth. However, little is known about the impact of the ACE/DD genotype and D allele dose on the cardiac changes in adult endurance athletes. METHODS; Echocardiographic studies (including tissue Doppler) were performed in 61 male endurance athletes ranging in age from 25 to 40 years, with a similar period of training (15.6 +/- 4 h/week for 12.6 +/- 5.7 years). The ACE genotype (insertion [I] or deletion [D] alleles) was ascertained by polymerase chain reaction (DD = 27, ID = 31, and II = 3). Athletes with the DD genotype were compared with their ID counterparts. RESULTS: The DD genotype was associated with a higher left ventricular mass index (LVMI) than the ID genotype (162.6 +/- 36.5 g/m(2) vs. 141.6 +/- 34 g/m(2), p = 0.031), regardless of other confounder variables. As a result, 70.4% of DD athletes and only 42% of ID athletes met the criteria for LVH (p = 0.037). Although systolic and early diastolic myocardial velocities were similar in DD and ID subjects, a more prolonged E-wave deceleration time (DT) was observed in DD as compared with ID athletes, after adjusting for other biologic variables (210 +/- 48 ms vs. 174 +/- 36 ms, respectively; p = 0.008). Finally, a positive association between DT and myocardial systolic peak velocity (medial and lateral peak S(m)) was only observed in DD athletes (p = 0.013, r = 0.481). CONCLUSIONS: The ACE/DD genotype is associated with the extent of exercise induced LVH in endurance athletes, regardless of other known biologic factors. PMID- 12906985 TI - Neo-aortic root dilation and valve regurgitation up to 21 years after staged reconstruction for hypoplastic left heart syndrome. AB - OBJECTIVES: We sought to assess the prevalence and progression of neo-aortic root dilation and valvar regurgitation after staged reconstruction for hypoplastic left heart syndrome (HLHS). BACKGROUND: In HLHS, the pulmonary valve functions as the neo-aortic valve. Neo-aortic valve dysfunction has been observed after arterial switch operation and the Ross procedure. METHODS: Patients with HLHS born before January 1995 who had the Fontan operation and had serial echocardiograms were included. Echocardiograms were reviewed preoperatively, after each surgical reconstruction, and at most recent follow-up for neo-aortic root size and severity of neo-aortic regurgitation (AR). Potential risk factors for neo-aortic valve dysfunction were assessed. RESULTS: Fifty-three patients met inclusion criteria. Bidirectional superior cavopulmonary anastomosis as an interim procedure was performed in 39 patients (74%). Median duration of follow up was 9.2 (range 5.1 to 21) years. During follow-up, the neo-aortic root progressively dilated out of proportion to body size over time, with 52 patients (98%) having a Z-score >2 at most recent follow-up. Neo-AR was present in 61% of patients at most recent follow-up, with progression over time in 26 patients (49%). However, neo-AR was more than mild in only three patients. Significantly larger neo-aortic root Z-scores were observed in patients with any degree of neo AR at most recent follow-up. No other anatomic or clinical variables correlated with severity of neo-AR or root dilation. CONCLUSIONS: After staged reconstruction for HLHS, neo-aortic root dilation and neo-AR progress over time. Early volume unloading does not have a beneficial impact on dilation of the neo aortic root. These findings raise concerns about neo-aortic valve function into adulthood. PMID- 12906986 TI - Long-term treatment with low-dose, but not high-dose, guanethidine improves ventricular function and survival of rats with heart failure after myocardial infarction. AB - OBJECTIVES: We sought to evaluate the effects of various doses of guanethidine, a sympathoinhibitory drug, on ventricular function and survival in chronic heart failure (CHF) after myocardial infarction (MI) in rats. BACKGROUND: Direct inhibition of sympathetic outflow by a sympathoinhibitory drug might be an effective approach to therapy of CHF. However, recent clinical trials suggest that excessive suppression of sympathetic activity has an adverse effect on outcome. It remains unclear whether the beneficial effects of the sympathoinhibitory drug would be modified by its dosage. METHODS: Three doses of guanethidine (low-dose [LG], 1 mg/kg/day; medium-dose, 3 mg/kg/day; high-dose, 10 mg/kg/day) were administered via an osmotic mini-pump for 4 weeks. Hemodynamics, left ventricular (LV) diameters, plasma and myocardial norepinephrine (NE) levels, and survival were determined for four weeks after MI. RESULTS: As compared with MI rats receiving vehicle, LG suppressed LV dilation (9.2 +/- 0.9 mm vs. 11.0 +/- 0.8 mm, p < 0.05) and improved LV fractional shortening (25.0 +/- 4.5% vs. 16.4 +/- 4.7%, p < 0.05) in association with a reduction of plasma NE levels (520 +/- 250 pg/ml vs. 1,000 +/- 570 pg/ml, p < 0.05), but not with a significant reduction of noninfarcted myocardial NE levels (154 +/- 71 ng/g vs. 207 +/- 71 ng/g). Low-dose guanethidine reduced 24-h (6%) and 28-day mortality (6%), as compared with untreated MI rats (36% and 52%, respectively). High-dose guanethidine also reduced 24-h mortality (12%) but increased 28-day mortality (91%), in association with a depletion of myocardial NE. Medium-dose guanethidine had no beneficial effects on LV hemodynamics or long-term survival. CONCLUSIONS: These results indicate that the dosage of the sympathoinhibitory drug might be quite important for the treatment of CHF. PMID- 12906987 TI - Sympatholysis and cardiac sympathetic nerve function in the treatment of congestive heart failure. PMID- 12906988 TI - Visualization of risk-area myocardium as a high-intensity, hyperenhanced "hot spot" by myocardial contrast echocardiography following coronary reperfusion: quantitative analysis. AB - OBJECTIVES: We examined whether delayed post-injection imaging of a new ultrasound contrast agent (BR-14) could produce prolonged opacification and hyperenhancement of myocardium subjected to coronary occlusion/reperfusion. BACKGROUND: We hypothesized that ultrasound exposure destroyed BR-14 and eliminated visualization of sustained myocardial opacification from retained microbubbles. METHODS: We studied eight open-chest dogs with 3 h of left anterior descending coronary artery (LAD) occlusion followed by 3 h of reperfusion. Myocardial contrast echocardiography (MCE) was performed before occlusion and 120 min after the onset of both occlusion and reperfusion. Ultrasound imaging was initiated 15 min after injection. Myocardial blood flow (MBF) was assessed by microspheres. RESULTS: Pre-occlusion images revealed uniform opacification of left ventricular myocardium greater than that of the cavity, with a mean intensity of the LAD bed of 8.66 +/- 1.38 dB. During occlusion, MCE resulted in the appearance of a perfusion defect in the LAD risk area (intensity 2.08 +/- 1.10 dB). After 120 min of reperfusion, the LAD risk-area myocardium manifested dense opacification of a higher intensity ("hot spot") than baseline (13.7 vs. 8.7 dB), but with reduced MBF consistent with accumulation of a high concentration of microbubbles. Increased MCE intensity was associated with a greater myeloperoxidase score. CONCLUSIONS: These data establish that contrast opacification by BR-14 may be selectively retained within the perfusion bed of a coronary artery subjected to occlusion/reperfusion. Such opacification exhibits defects with occlusion, manifests hyperenhanced intensity (hot spot) with reperfusion, is associated with the level of myeloperoxidase activity, and conforms to the area of myocardium subjected to altered flow. PMID- 12906989 TI - Intra-ventricular resynchronization for optimal left ventricular function during pacing in experimental left bundle branch block. AB - OBJECTIVES: We sought to investigate to what extent intra-ventricular asynchrony (intraVA) and inter-ventricular asynchrony (interVA) determine left ventricular (LV) function in canine hearts with left bundle branch block (LBBB) during ventricular pacing. BACKGROUND: Pacing therapy improves LV pump function in patients with heart failure and abnormal ventricular conduction supposedly due to resynchronization. However, the relationship between LV pump function and measures of asynchrony is not well established. METHODS: In 15 experiments, LV (various sites) and biventricular (BiV) pacing was performed at atrioventricular (AV) delays of 20 to 140 ms. Measured were the maximum rate of increase (dP/dt(max)) of LV pressure and LV stroke work (SW) (conductance catheter), interVA (time delay between the upslope of LV and RV pressures), and intraVA (from endocardial electrical activation maps). RESULTS: Induction of LBBB increased interVA (-6.4 +/- 8.6 to -28.4 +/- 8.5 ms [RV earlier]) and intraVA (4.9 +/- 2.4 to 18.0 +/- 3.3 ms), whereas LV dP/dt(max) and SW decreased (-13 +/- 18% and -39 +/- 24%, respectively). During LBBB, LV and BiV pacing increased LV dP/dt(max) and SW (mean increases 14% to 21% and 11% to 15%, respectively) without changing diastolic function or preload. Optimal improvement in LV function was obtained consistently when intraVA returned to pre-LBBB values, while interVA remained elevated. Normalization of intraVA required AV delays shorter than the baseline PQ time during LV apex and BiV pacing, thus excluding endogenous LV activation, but AV delays virtually equal to the baseline PQ time (difference 4 +/- 9 ms, p = NS) during pacing at (mid)lateral LV sites to obtain fusion between pacing-induced and endogenous activation. CONCLUSIONS: In LBBB hearts, optimal restoration of LV systolic function by pacing requires intra ventricular resynchronization. The optimal AV delay to achieve this depends on both the site of pacing and baseline PQ time. PMID- 12906990 TI - Triphasic waveforms are superior to biphasic waveforms for transthoracic defibrillation: experimental studies. AB - OBJECTIVES: Our objective was to evaluate the efficacy of triphasic waveforms for transthoracic defibrillation in a swine model. BACKGROUND: Triphasic shocks have been found to cause less post-shock dysfunction than biphasic shocks in chick embryo studies. METHODS: After 30 s of electrically induced ventricular fibrillation (VF), each pig in part I (n = 32) received truncated exponential biphasic (7.2/7.2 ms) and triphasic (4.8/4.8/4.8 ms) transthoracic shocks. Each pig in part II (n = 14) received biphasic (5/5 ms) and triphasic shocks (5/5/5 ms). Three selected energy levels (50, 100, and 150 J) were tested for parts I and II. Pigs in part III (n = 13) received biphasic (5/5 ms) and triphasic (5/5/5 ms) shocks at a higher energy (200 and 300 J). Although the individual pulse durations of these shocks were equal, the energy of each pulse varied. Nine pigs in part I also received shocks where each individual pulse contained equal energy but was of a different duration (biphasic 3.3/11.1 ms; triphasic 2.0/3.2/9.2 ms). RESULTS: Triphasic shocks of equal duration pulses achieved higher success than biphasic shocks at delivered low energies: <40 J: 38 +/- 5% triphasic vs. 19 +/- 4% biphasic (p < 0.01); 40 to <50 J: 66 +/- 7% vs. 42 +/- 7% (p < 0.01); and 50 to <65 J: 78 +/- 4% vs. 54 +/- 5% (p < 0.05). Shocks of equal energy but different duration pulses achieved relatively poor success for both triphasic and biphasic waveforms. Shock-induced ventricular tachycardia (VT) and asystole occurred less often after triphasic shocks. CONCLUSIONS: Triphasic transthoracic shocks composed of equal duration pulses were superior to biphasic shocks for VF termination at low energies and caused less VT and asystole. PMID- 12906991 TI - Pinhole single-photon emission computed tomography for myocardial perfusion imaging of mice. AB - OBJECTIVES: Although transgenic mice have emerged as powerful experimental models of cardiovascular disease, methods for in vivo phenotypic assessment and characterization remain limited, motivating the development of new instruments for biologic measurement. BACKGROUND: We have developed a single-photon emission computed tomography system with a pinhole collimator (pinhole SPECT) for high resolution cardiovascular imaging of mice. In this study, we describe a protocol for myocardial perfusion imaging of mice using technetium-99m ((99m)Tc)-sestamibi and demonstrate the feasibility for measurement of perfusion defect size from pinhole SPECT images. METHODS: Mice were anesthetized and injected with 370 MBq (10 mCi) of (99m)Tc-sestamibi. Tomographic projection images were acquired by rotating each mouse in a vertical axis in front of a stationary clinical scintillation camera equipped with a pinhole collimator. BALB/c mice (n = 15) were imaged after the permanent ligation of the left anterior descending coronary artery. The resulting defect size was measured from circumferential profiles of short-axis images. After imaging, the hearts were excised and sectioned to obtain ultra-high resolution digital autoradiographs of (99m)Tc-sestamibi, from which the actual infarct size was determined. RESULTS: Reconstructed image quality was equivalent to that obtained for clinical myocardial perfusion imaging. Linear regression analysis produced a correlation coefficient of 0.83 (p < 0.001) between the measured and actual values of the defect size. CONCLUSIONS: These results demonstrate that myocardial perfusion can be characterized qualitatively and quantitatively in mice using pinhole SPECT. PMID- 12906992 TI - Reflections on the first year. PMID- 12906993 TI - President's page: Counting heads, coming up short: the ratio of cardiologists to patients is becoming inadequate. PMID- 12906994 TI - Anabolic steroid-induced echocardiographic characteristics of professional football players? PMID- 12906996 TI - Cellular myocardial reconstruction using human myoblasts. PMID- 12907000 TI - Is science stuck in the Middle Ages? PMID- 12907001 TI - Predictive cancer genomics--what do we need? PMID- 12907002 TI - (Almost) three cheers for UK genetics White Paper. PMID- 12907003 TI - Neuroprotection and the role of neuroglobin. PMID- 12907004 TI - PowerPoint: shot with its own bullets. PMID- 12907005 TI - End-of-life decision-making in six European countries: descriptive study. AB - BACKGROUND: Empirical data about end-of-life decision-making practices are scarce. We aimed to investigate frequency and characteristics of end-of-life decision-making practices in six European countries: Belgium, Denmark, Italy, the Netherlands, Sweden, and Switzerland. METHODS: In all participating countries, deaths reported to death registries were stratified for cause (apart from in Switzerland), and samples were drawn from every stratum. Reporting doctors received a mailed questionnaire about the medical decision-making that had preceded the death of the patient. The data-collection procedure precluded identification of any of the doctors or patients. All deaths arose between June, 2001, and February, 2002. We weighted data to correct for stratification and to make results representative for all deaths: results were presented as weighted percentages. FINDINGS: The questionnaire response rate was 75% for the Netherlands, 67% for Switzerland, 62% for Denmark, 61% for Sweden, 59% for Belgium, and 44% for Italy. Total number of deaths studied was 20480. Death happened suddenly and unexpectedly in about a third of cases in all countries. The proportion of deaths that were preceded by any end-of-life decision ranged between 23% (Italy) and 51% (Switzerland). Administration of drugs with the explicit intention of hastening death varied between countries: about 1% or less in Denmark, Italy, Sweden, and Switzerland, 1.82% in Belgium, and 3.40% in the Netherlands. Large variations were recorded in the extent to which decisions were discussed with patients, relatives, and other caregivers. INTERPRETATION: Medical end-of-life decisions frequently precede dying in all participating countries. Patients and relatives are generally involved in decision-making in countries in which the frequency of making these decisions is high. PMID- 12907006 TI - US health-care system faces cost and insurance crises. Rising costs, growing numbers of uninsured, and quality gaps trouble world's most expensive health-care system. PMID- 12907007 TI - Decreased mortality of ischaemic heart disease among carriers of haemophilia. AB - BACKGROUND: Coagulation plays an important part in ischaemic cardiovascular disease. Results of studies have shown that extremes in hypocoagulability protect against ischaemic cardiovascular disease. We have investigated overall mortality and death from cardiovascular causes in carriers of haemophilia, who in most cases have mildly decreased coagulability without clinical signs. METHODS: We followed-up a cohort of 1012 mothers of all known people with haemophilia in the Netherlands from birth to death, or the end-of-study date (41984 person years of follow-up). We obtained vital status and causes of death, if deceased, and compared overall and cause-specific mortality in our cohort with that in the general Dutch female population adjusted for age and calendar period by calculating the standardised mortality ratio (SMR). FINDINGS: Overall mortality was reduced by 22% (261 observed deaths, 333.74 expected; SMR 0.78 [95% CI 0.69 0.88]). Deaths from ischaemic heart disease were reduced by 36% (39 observed deaths, 60.53 expected; SMR 0.64 [0.47-0.88]). We did not note decreased mortality for cerebral stroke (ischaemic and haemorrhagic combined) (28 observed deaths, 36.82 expected; SMR 0.76 [0.53-1.10]). A separate analysis of these two types of stroke was not possible. Women in our cohort had an increased risk of death from extracranial haemorrhage (5 observed deaths, 0.18 expected; SMR 27.78 [8.49-58.18]); however, the number of deaths from this cause was much lower than that for ischaemic heart disease. CONCLUSION: The results show that a mild decrease in coagulability has a protective effect against fatal ischaemic heart disease. PMID- 12907008 TI - Efficacy and safety of seven-valent conjugate pneumococcal vaccine in American Indian children: group randomised trial. AB - BACKGROUND: Streptococcus pneumoniae is the main cause of invasive bacterial disease in children aged younger than 2 years. Navajo and White Mountain Apache children have some of the highest rates of invasive pneumococcal disease documented in the world. We aimed to assess the safety and efficacy of a seven valent polysaccharide protein conjugate pneumococcal vaccine (PnCRM7) against such disease. METHODS: In a group-randomised study, we gave this vaccine to children younger than 2 years from the Navajo and White Mountain Apache Indian reservations; meningococcal type C conjugate vaccine (MnCC) served as the control vaccine. Vaccine schedules were determined by age at enrollment. We recorded episodes of invasive pneumococcal disease and serotyped isolates. Analyses were by intention to treat and per protocol. FINDINGS: 8292 children enrolled in the trial. In the per protocol analysis of the primary efficacy group (children enrolled by 7 months of age) there were eight cases of vaccine serotype disease in the controls and two in the PnCRM7 group; in the intention-to-treat analysis we noted 11 cases of vaccine serotype disease in the MnCC control group and two in the PnCRM7 group. After group randomisation had been controlled for, the per protocol primary efficacy of PnCRM7 was 76.8% (95% CI -9.4% to 95.1%) and the intention-to-treat total primary efficacy was 82.6% (21.4% to 96.1%). INTERPRETATION: PnCRM7 vaccine prevents vaccine serotype invasive pneumococcal disease even in a high risk population. Other regions with similar disease burden should consider including this vaccine in the routine childhood vaccine schedule. PMID- 12907009 TI - Gene expression profiling for the prediction of therapeutic response to docetaxel in patients with breast cancer. AB - BACKGROUND: Systemic chemotherapy for operable breast cancer substantially decreases the risk of death. Patients often have de novo resistance or incomplete response to docetaxel, one of the most active agents in this disease. We postulated that gene expression profiles of the primary breast cancer can predict the response to docetaxel. METHODS: We took core biopsy samples from primary breast tumours in 24 patients before treatment and then assessed tumour response to neoadjuvant docetaxel (four cycles, 100 mg/m2 daily for 3 weeks) by cDNA analysis of RNA extracted from biopsy samples using HgU95-Av2 GeneChip. FINDINGS: From the core biopsy samples, we extracted sufficient total RNA (3-6 microg) for cDNA array analysis using HgU95-Av2 GeneChip. Differential patterns of expression of 92 genes correlated with docetaxel response (p=0.001). Sensitive tumours had higher expression of genes involved in cell cycle, cytoskeleton, adhesion, protein transport, protein modification, transcription, and stress or apoptosis; whereas resistant tumours showed increased expression of some transcriptional and signal transduction genes. In leave-one-out cross-validation analysis, ten of 11 sensitive tumours (90% specificity) and 11 of 13 resistant tumours (85% sensitivity) were correctly classified, with an accuracy of 88%. This 92-gene predictor had positive and negative predictive values of 92% and 83%, respectively. Correlation between RNA expression measured by the arrays and semiquantitative RT-PCR was also ascertained, and our results were validated in an independent set of six patients. INTERPRETATION: If validated, these molecular profiles could allow development of a clinical test for docetaxel sensitivity, thus reducing unnecessary treatment for women with breast cancer. PMID- 12907010 TI - Blunt chest trauma producing acute myocardial infarction in a rugby player. PMID- 12907011 TI - Zoonotic transmission of hepatitis E virus from deer to human beings. AB - Zoonosis has been suggested for hepatitis E virus (HEV) infection, but so far is based only on indirect evidence. We experienced a series of cases of HEV infection among people who had eaten uncooked deer meat 6-7 weeks before. On testing, a left over portion of the deer meat, kept frozen to eat in the future, was positive for HEV RNA, whose nucleotide sequence was identical to those from the patients. Patients' family members who ate none or very little of the deer meat remained uninfected. These findings provide direct evidence for HEV infection to be a zoonosis. PMID- 12907012 TI - Chromoendoscopy with methylene blue and associated DNA damage in Barrett's oesophagus. AB - Chromoendoscopy with methylene blue has been proposed to improve targeting of biopsies to specialised intestinal metaplasia and dysplasia in Barrett's oesophagus. However, methylene blue can induce oxidative damage of DNA when photosensitised by white light. We show that damage to DNA is increased in Barrett's mucosa after chromoendoscopy with methylene blue, an effect apparently dependent on presence of both methylene blue and endoscopic white light. Exposure of Barrett's mucosa to DNA damage during endoscopy warrants caution since it could accelerate carcinogenesis. This risk needs to be carefully balanced against the possible benefit of improved early detection of preneoplastic lesions with methylene blue chromoendoscopy. PMID- 12907013 TI - Coeliac disease. AB - Coeliac disease is a genetically-determined chronic inflammatory intestinal disease induced by an environmental precipitant, gluten. Patients with the disease might have mainly non-gastrointestinal symptoms, and as a result patients present to various medical practitioners. Epidemiological studies have shown that coeliac disease is very common and affects about one in 250 people. The disease is associated with an increased rate of osteoporosis, infertility, autoimmune diseases, and malignant disease, especially lymphomas. The mechanism of the intestinal immune-mediated response is not completely clear, but involves an HLA DQ2 or HLA-DQ8 restricted T-cell immune reaction in the lamina propria as well as an immune reaction in the intestinal epithelium. An important component of the disease is the intraepithelial lymphocyte that might become clonally expanded in refractory sprue and enteropathy-associated T-cell lymphoma. Study of the mechanism of the immune response in coeliac disease could provide insight into the mechanism of inflammatory and autoimmune responses and lead to innovations in treatment. PMID- 12907014 TI - Right ventricular involvement in myocardial infarction and cardiogenic shock. AB - CONTEXT: Right ventricular involvement in acute myocardial infarction and cardiogenic shock has received little attention by clinicians and researchers, although its pathophysiology, clinical presentation, and natural history are distinctly different from those of left ventricular infarction and associated cardiogenic shock. Right ventricular shock has important therapeutic implications for the management of patients, which need to be recognised. STARTING POINT: Investigators at the SHOCK Registry (Alice Jacobs and colleagues, J Am Coll Cardiol 2003; 341: 1273-79) evaluated 49 patients with cardiogenic shock predominantly due to right ventricular infarction and compared them with 884 patients with cardiogenic shock and predominantly left ventricular failure. Perhaps surprisingly, these investigators found that the in-hospital mortality of patients with right ventricular shock was not significantly lower than that of patients with left ventricular shock (53% vs 61%, p=0.296), despite the fact that patients with right ventricular shock were younger, with a lower prevalence of previous infarctions, fewer anterior infarct locations, and less multivessel disease. There was a shorter median time between index infarction and diagnosis of shock in patients with right ventricular shock. In multivariate analysis, right ventricular shock was not an independent predictor of lower in-hospital mortality. WHERE NEXT? The unexpectedly high mortality of patients with cardiogenic shock due to predominantly right ventricular infarction challenges the general notion that right ventricular involvement in myocardial infarction has only little relevance for patient's outcome. Therefore, more attention should be given to the detection of right ventricular involvement in acute myocardial infarction and particularly in cardiogenic shock. If right ventricular shock is diagnosed, urgent reperfusion of the infarct related artery and appropriate circulatory support are required. PMID- 12907015 TI - Euthanasia and other end-of-life decisions in the Netherlands in 1990, 1995, and 2001. AB - Empirical data on the rate of euthanasia, physician-assisted suicide, and other end-of-life decisions have greatly contributed to the debate about the role of such practices in modern health care. In the Netherlands, the continuing debate about whether and when physician-assisted dying is acceptable seems to be resulting in a gradual stabilisation of end-of-life practices. We replicated interview and death-certificate studies done in 1990 and 1995 to investigate whether end-of-life practices had altered between 1995 and 2001. Since 1995, the demand for physician-assisted death has not risen among patients and physicians, who seem to have become somewhat more reluctant in their attitude towards this practice. PMID- 12907016 TI - Beyond release: the equitable use of genomic information. PMID- 12907017 TI - Osteochondral graft transplantation for steroid-induced osteonecrosis of the femoral condyle. PMID- 12907018 TI - Employer struggles with rising health costs. Fewer benefits and employees must contribute more. PMID- 12907019 TI - Cochrane and the salt debate. PMID- 12907020 TI - The place for politics? PMID- 12907021 TI - New generation versus conventional antipsychotics. PMID- 12907022 TI - Insurance loss leaves doctors scrambling for coverage. Insurer says it can no longer cover high-risk specialties. PMID- 12907023 TI - Greece is iodine sufficient. PMID- 12907024 TI - SARS genome patent: to manage and to share. PMID- 12907025 TI - Panspermia--true or false? PMID- 12907026 TI - Panspermia--true or false? PMID- 12907027 TI - US doctors press for malpractice reform . But lawyers and consumer groups say proposals unfair to patients. PMID- 12907028 TI - Panspermia--true or false? PMID- 12907029 TI - Spiritual wellbeing and care of terminally ill patients. PMID- 12907030 TI - Fossil record of meningioma. PMID- 12907032 TI - Research issues in CPD. PMID- 12907033 TI - Limits imposed on residents' work hours. Reduced workload said to increase patient--and resident--safety. PMID- 12907034 TI - Addiction myths. PMID- 12907035 TI - Antibiotics from the sky. PMID- 12907047 TI - The prevalence of neurosensorial hearing loss among cystic fibrosis patients from Hospital de Clinicas de Porto Alegre. AB - Cystic fibrosis (CF) is considered one of the most prevalent lethal genetic disorders. The life expectancy of those patients has been rising. Besides the well-known prevalence of chronic rhinosinusitis, neurosensory hearing loss (NSHL) due to the large use of ototoxic antibiotics may be another problem that should receive special attention in otolaryngology (ENT). OBJECTIVES: Define retrospectively the prevalence of NSHL among patients with CF from the Hospital de Clinicas de Porto Alegre (HCPA) and based on this data discuss the need for a specific out patient ENT-CF clinic. MATERIAL AND METHOD: Files from CF HCPA patients were reviewed for age at diagnosis, the current age, types of antibiotics used, number of treatments, percentage with audiologic evaluation and its respective results. RESULTS: One hundred and seven files were reviewed. The mean age at diagnosis was 1.33 years old (SD 1.97, range 0-12). The mean age of the sample was 7.87 (SD 4.49, range 1-22). Audiologic evaluation was performed in 39.3% (42) of the patients. 28.56% (12) had some degree of NSHL, while 36.3% did not present acoustic reflex. From 667 treatments with antibiotics, 49.5% were with tobramycin, 43.4% with amycacin and 7% with gentamicin. CONCLUSIONS: The prevalence of NSHL among CF patients supports the need for a specific ENT-CF outpatient clinic. This clinic would help on the prevention and treatment of NSHL due to the use of ototoxic antibiotics and on the better understanding/control of rhinosinusal disorders. PMID- 12907048 TI - Effect of positional change and inhalant anesthesia on parameters of acoustic reflectometry. AB - OBJECTIVE: Purpose of this study was to find out the effect of positional change and inhalant anesthesia on acoustic reflectometry (AR) parameters (reflectivity and curve angle). METHOD: AR parameters were measured on 58 ears with otitis media in sitting position before anesthesia and in supine position under inhalant anesthesia, subsequently. RESULTS: Under anesthesia, ears with effusion disclosed more changes in reflectivity (Chi-squared analysis, chi2-test; P<0.05) and curve angle (P>0.1) than those without effusion. Further, inhalant anesthesia caused more changes in the false negative ears (63.63%) than in those with effusion having positive test before anesthesia (12.90%) (P<0.001). CONCLUSIONS: From the data of this study, it could be said that reflectivity shows changes according to the amount of effusion which is in contact with the tympanic membrane under anesthesia, and that curve angle becomes more sensitive to detect effusion when anesthetic gas diffuses into the middle ear with effusion, probably due to the pushing of effusion towards the tympanic membrane. PMID- 12907049 TI - A comparison between parents of children with cochlear implants and parents of children with hearing aids regarding parental distress and treatment expectations. AB - OBJECTIVE: The aim of the present study was to compare the parents of children with a hearing aid (HA) and children with a cochlear implant (CI) regarding their psychological distress, their expectations from treatment, their family climate, and the way they first obtained information on HA/CI. METHODS: 154 parents (return quota 41%; 81 mothers and 73 fathers) of 90 children with a HA and 103 parents (return quota 59%; 57 mothers and 46 fathers) of 57 children with a CI were interviewed by means of a questionnaire. RESULTS: Both groups of parents felt distressed, particularly at the time of diagnosis. Their psychological well being was gradually stabilized in the further course of rehabilitation. Due to the operation associated with it, fitting with a CI brought on a phase of heightened parental psychological distress compared with less invasive treatment with a HA. Regarding family climate, more distress was found in parents of CI children than in parents of HA children. Expectations from therapy appeared realistic in both parental groups; however, after CI fitting, the parents of the CI children showed heightened expectations by comparison with the parents of the HA children. CONCLUSIONS: The results of our study suggest that the parents of hearing impaired children fitted with a HA or a CI may be divided into two subgroups with divergent psychosocial parameters. For the counseling of the parents of hearing impaired children in clinical practice, it would seem important to take these specific differences into consideration. PMID- 12907050 TI - Bronchoscopy induces intestinal mucosal barrier dysfunction: a possible role for nitric oxide. AB - OBJECTIVE: This study investigates the effect of bronchoscopy on intestinal mucosal barrier function and its association with intestinal nitric oxide production. METHODS: 30 rats were used. The study group (n=15) underwent rigid bronchoscopy. At 24 h following bronchoscopy, ileal nitrite/nitrate levels were evaluated. The ileum was also examined for mucosal damage, and graded according Chiu's histologic injury scale. RESULTS: In the bronchoscopy group, the ileal nitrite/nitrate levels were significantly higher than those of controls (398.5 +/ 85.1 and 44.5 +/- 6.6 nmol/g tissue, respectively, P=0.001). In the bronchoscopy group, the mucosal damage was significant, compared with those of controls (mean ranks, 22.8 and 8.2, P<0.0001). The changes varied from denuded villi and dilated capillaries to significant architectural distortion, lamina propria disintegration, ulceration and hemorrhage. Significant correlation was found between ileal nitrite/nitrate levels and mucosal damage in the bronchoscopy group (rs=0.56, P=0.03). CONCLUSION: This study suggests that bronchoscopy induces intestinal mucosal barrier dysfunction in association with excess intestinal nitric oxide production. These events may be involved in mechanisms responsible for bacterial translocation after bronchoscopy. PMID- 12907051 TI - Penetration of cefprozil to middle ear effusion in children with chronic otitis media with effusion. AB - BACKGROUND: Because of chronic otitis media with effusion (COME) demonstrates pathogenic bacteria, treatment with appropriate antibiotic is reasonable. OBJECTIVE: We determined the penetration of cefprozil into the middle ear effusion (MEE) in children with COME. MATERIALS AND METHODS: 25 patients 2-13 years of age with COME were eligible for study. After the single dose of 15 mg/kg patients were assigned to have MEE, and serum samples were obtained during ventilation tube insertion at 0.5, 2, 3, 5, or 6 h after administration of the dose. The concentration of cefprozil was measured using validated high performance liquid chromatography method. RESULTS: The mean concentrations of cefprozil in the MEE ranged from 0.4 to 4.4 microg/ml. The penetration into MEE was rapid and effective. Cefprozil in the MEE was maintained at a greater level than MIC 90 in S. pneumoniae for at least 6 h after administration of 15 mg/kg. CONCLUSION: Cefprozil penetrated well into the MEE in children with COME. PMID- 12907052 TI - Epidemiology of hearing impairment at three Flemish Institutes for Deaf and Speech Defective Children. AB - A retrospective analysis of 190 records of hearing impaired children up to the age of 14, all educated at one of three Flemish Institutes for Deaf and Speech Defective Children in Belgium, was performed. Until 1999, the well-known behavioral test (Ewing test) was used in the Flemish national screening program for hearing losses. Because it presented a lot of disadvantages, it has been replaced by Automated Auditory Brainstem Responses (AABR), enabling the Flemish national neonatal screening program to commence. This study is an extension of the preliminary results of a recently performed retrospective analysis in one Flemish institute of the hearing loss of patients that were diagnosed in the pre AABR era. The authors analyzed the following data: etiology, risk factors of congenital hearing impairment, the patient's history from the moment of the first suspicion to diagnosis and treatment. The median age of the children was 8.5 years (2-14 year). In 66.5% the parents and/or grandparents were the first to raise suspicion, this was at a median age of approximately 9 months. The diagnosis was often made late, at a median age of 15 months (0-88 months). The etiology was unknown in 32.6% of the cases. The only cast-iron certain diagnoses were pre- and perinatal infections, syndromal and genetic hearing loss, and acquired infections (meningitis and measles). No risk factors of hearing loss, as they are stated by the Joint Committee on Infant Hearing, were found in more than 50% of the cases. Audiometry was performed in all cases, often supplemented with Auditory Brainstem Responses and/or click-evoked otoacoustic emissions, while other diagnostic investigations (imaging, genetics, etc.) were only variably performed. Finally, the authors confirm the need for universal neonatal screening, which only recently started in Belgium, and suggest that a detailed protocol should be established to pursue a coherent diagnostic policy. PMID- 12907053 TI - Esophagoscopy for removal of foreign bodies in the pediatric population. AB - A retrospective chart review of children who had rigid esophagoscopy for potential foreign body ingestion from 1998 to 2001 was conducted at Children's Hospital of Michigan Detroit. All pediatric patients less than 11 years of age who presented with suspected foreign body ingestion in a hospital setting over a 4-year period were retrospectively studied. Patient characteristics noted included age, sex and clinical presentation. Pre-operative radiographic findings, esophagoscopy findings, clinical presentations and types of foreign bodies were recorded. A history compatible with foreign body ingestion dictates diagnostic endoscopy with or without radiographic confirmation, and an acceptable rate of performing esophagoscopy without finding a foreign body was found to be 6.2%. PMID- 12907054 TI - Tonsillectomy--clinical consequences twenty years after surgery? AB - Tonsillectomy (T) is one of the most common surgical procedures performed on children. Long-term follow-up studies concerning its consequences are lacking. This study is the first study done on a group of patients that underwent T in their childhood, about 20 years ago. The investigation is a cohort study, which followed-up 18 patients who were tonsillectomized 20 years ago. It was to be determined whether these subjects suffer from more respiratory tract infections (or other infections) today, than people who are not tonsillectomized. A group of 54 age-matched subjects were selected for comparison. A questionnaire was mailed to the study population. No significant differences were found between the groups in the frequency of upper respiratory tract infection (URI). The mean number of URI's was approximately [MSOffice1]2.5 per year in both groups. The duration of the URI's was identical in each group. A high temperature was present to the same extent in each group. Absence from work, number of visits to physicians and the use of antibiotics were the same in each group. However, the prevalence of chronic disease was greater in the T-group than in the comparison group. The difference was significant with a Relative Risk of 9.41 and a Confidence Interval differing from 1 (1.13/=40 years, and stratified by parity, were calculated by logistic regression. RESULTS: In nulliparas, labor induction was associated with an increase in cesarean delivery from 13.7% to 24.7% (adjusted odds ratio [OR] 1.70; 95% confidence interval [CI] 1.48, 1.95]). In multiparas, induction was associated with an increase from 2.4% to 4.5% (OR 1.49; 95% CI 1.10, 2.00). Other variables that placed a nulliparous woman at increased risk for cesarean delivery included maternal age of at least 35 years and gestational ages over 40 weeks. For multiparas, only maternal age 40 years or older and gestational age of 41 weeks were associated with an increase in cesarean deliveries. CONCLUSION: Induction of labor, older maternal age, and gestational age over 40 weeks each independently increase the risk for cesarean delivery in both nulliparous and multiparous women. Although the relative risk from induction is similar in nulliparas and multiparas, the absolute magnitude of the increase is much greater in nulliparas (11% versus 2.1%). PMID- 12907102 TI - Anticardiolipin and anti-beta2-glycoprotein-I antibodies in preeclampsia. AB - OBJECTIVE: To estimate whether antiphospholipid antibodies, specifically anticardiolipin and anti-beta(2)-glycoprotein-I antibodies, are associated with preeclampsia. METHODS: Plasma was prospectively obtained from four groups of pregnant women: those with 1) mild preeclampsia (n = 109); 2) severe preeclampsia (n = 134); 3) hemolysis, elevated liver enzymes, low platelets (HELLP) syndrome (n = 57); and 4) normotensive controls (n = 100). Anticardiolipin and anti beta(2)-glycoprotein-I levels were determined by enzyme-linked immunoassay. RESULTS: Subjects with mild preeclampsia, severe preeclampsia, and HELLP syndrome did not have significantly elevated levels of immunoglobulin G (IgG) and IgM anticardiolipin and anti-beta(2)-glycoprotein-I antibodies compared with normotensive controls (P >.05, Kruskal-Wallis). Similarly, subjects with mild preeclampsia, severe preeclampsia, and HELLP syndrome did not have a significantly higher proportion of women testing positive for each autoantibody compared with normotensive controls (chi(2)). The proportion of patients testing positive for anticardiolipin and anti-beta(2)-glycoprotein-I antibodies were similar in patients with preeclampsia developing before and after 34 weeks' gestation (chi(2)). CONCLUSION: Circulating levels of both anticardiolipin and anti-beta(2)-glycoprotein-I antibodies were not increased in patients with mild preeclampsia, severe preeclampsia, or HELLP syndrome compared with normotensive controls. Our data do not support routine testing for anticardiolipin and anti beta(2)-glycoprotein-I antibodies in women with preeclampsia. PMID- 12907103 TI - Total versus subtotal hysterectomy: a survey of gynecologists. AB - OBJECTIVE: To estimate the attitudes and practice of gynecologists in the Washington, DC, Maryland, and Virginia area regarding total versus subtotal abdominal hysterectomy. METHODS: A questionnaire with 18 questions on physicians' attitudes and practice regarding total versus subtotal hysterectomy was mailed to 1647 gynecologists in Washington, Maryland, and Virginia. RESULTS: The corrected response rate was 51.2%. Forty-five percent of respondents stated that they always removed the cervix. The most common reason cited was to eliminate the risk of cervical cancer. The most common reason for subtotal hysterectomy was surgical difficulty leading to an intraoperative change of procedure. Only 17.8% of respondents always counseled women regarding the advantages and disadvantages of both total and subtotal hysterectomy; 63% rarely or never did. Nineteen percent always offered women a choice between the procedures; 61% rarely or never did. Eighty-eight percent of respondents felt that the risk of cancer in the cervical stump was small or negligible. Gender of the physicians or year of completion of residency made no significant impact on patients being counseled about both procedures or being offered a choice between the two. CONCLUSION: Most gynecologists surveyed favor total abdominal hysterectomy over subtotal hysterectomy. Few counsel women regarding the options of total and subtotal hysterectomy or offer a choice between the procedures. Given that there are no convincing data proving the superiority of either procedure over the other, it may be reasonable to discuss the potential advantages and disadvantages of both procedures with women undergoing hysterectomy for benign disease and to offer them a choice. PMID- 12907104 TI - Risk of mesh erosion with sacral colpopexy and concurrent hysterectomy. AB - OBJECTIVE: To examine short- and long-term mesh-related complications in women undergoing abdominal sacral colpopexy with concurrent hysterectomy, compared with women with a prior hysterectomy undergoing sacral colpopexy alone. METHODS: Patient characteristics, hospital complications, postoperative clinical course, and long-term graft-related complications were reviewed for all women with genital prolapse who underwent abdominal sacral colopexy between 1996 and 1998. Women with concurrent hysterectomy were compared with women with vaginal prolapse after a prior hysterectomy. RESULTS: One hundred twenty-four patients, 60 with concurrent hysterectomy and 64 with prior hysterectomy, were observed postoperatively for a median of 35.5 (0-74) months. Demographics of the two groups were similar, with a mean age of 65.1 +/- 9.4 years and a mean body mass index of 25.8 +/- 4.2 kg/m(2). Eighty percent of colpopexies used prolene synthetic mesh and 20% allograft material. Initial operative and hospital complications were rare in both groups and included a blood transfusion of 2 U, a ureteral transection, a wound infection, heart block, and an arrhythmia. Delayed graft complications included one mesh erosion in a patient with a prior hysterectomy that was managed by office resection (0.8%). CONCLUSION: Concurrent hysterectomy with abdominal sacral colopopexy has a low incidence of mesh complications and can be used as a first-line treatment for genital prolapse. PMID- 12907105 TI - A randomized clinical trial of lorazepam for the reduction of postoperative urinary retention. AB - OBJECTIVE: To estimate whether lorazepam can decrease the incidence of postoperative urinary retention after ambulatory gynecologic surgeries. METHODS: A randomized clinical trial at an urban academic medical center was performed. Lorazepam (1 mg) or a placebo was administered intravenously 1 hour after ambulatory gynecologic procedures. Ninety patients were randomized to receive lorazepam (n = 47) or a placebo (n = 43). A computer-generated random number table was used for group assignment. Concealment was maintained by the hospital pharmacy, which prepared and dispensed the medication. Physicians, nurses, and patients were blinded to the group assignment. The time of the first spontaneous void and other information about the surgery and recovery were then recorded. RESULTS: There was no difference in the overall mean time to void between the lorazepam (3.1 +/- 1.2 hours) and placebo (2.8 +/- 2.0 hours) groups (P =.5). The time to void did not correlate with intravenous fluid use, length of procedure, type of procedure, narcotic use, body mass index, age, type of catheterization, or estimated blood loss. CONCLUSION: Medium half-life benzodiazepines do not significantly reduce the incidence of postoperative urinary retention. PMID- 12907106 TI - Does anovulation exist in eumenorrheic women? AB - OBJECTIVE: To determine whether anovulation exists in normally menstruating women. METHODS: In a database of 550 consecutive couples seeking pregnancy, results of the midluteal serum progesterone level analysis planned for 7 days before the onset of the next menses were examined in women with predictable cycles shorter than 35 days. RESULTS: Of the 550 couples seeking pregnancy, 410 of the female partners (74.5%) were eumenorrheic. Fifteen of these women (3.7%) had apparently anovulatory cycles with a progesterone lower than the normal ovulatory value of 15 nmol/L. Further examination showed that four of the 15 women (26.7%) had an isolated prolonged cycle, whereas an additional four (26.7%) failed to have their sample taken at an appropriate time. One (6.7%) had a low progesterone level that was normal in the subsequent cycle. Two patients (13.3%) were older than 40, both having elevated early follicular follicle-stimulating hormone levels. One patient (6.7%) conceived in the following menstrual cycle without further evaluation. The three remaining women (20%) showed consistently apparently anovulatory cycles. However, the levels were exclusively above the follicular range. CONCLUSION: Our findings cast doubt on the concept of anovulatory cycles in eumenorrheic women and suggest that further examination of the lower level of ovulatory progesterone may indeed be necessary. PMID- 12907107 TI - Increased plasma adrenomedullin in women with recurrent pregnancy loss. AB - OBJECTIVE: To evaluate vascular changes and uterine perfusion in women with recurrent pregnancy loss. METHODS: We measured plasma levels of adrenomedullin of 100 pregnant women in the midluteal phase of a nonpregnant cycle (control group: n = 62; recurrent pregnancy loss group: n = 38). We measured the pulsatility index (PI) in the uterine arteries by transvaginal pulsed Doppler ultrasonography at the same time. RESULTS: The plasma level of adrenomedullin in women with recurrent pregnancy loss (5.6 +/- 1.9, mean +/- standard deviation) was significantly higher (P >.001) than that in control women (3.6 +/- 1.7). Uterine arterial PI of women with recurrent pregnancy loss (2.70 +/- 0.47) was significantly higher (P >.001) than that in control women (2.09 +/- 0.39). Plasma level of adrenomedullin had a significant positive correlation with uterine arterial PI both in the control group (r =.58, P <.001) and in the recurrent pregnancy loss group (r =.78, P <.001). Both plasma adrenomedullin concentration (7.2 +/- 2.3) and uterine arterial PI (3.06 +/- 0.36) were significantly high in women with antiphospholipid antibodies. CONCLUSION: Plasma adrenomedullin may serve as a useful biochemical marker for recurrent pregnancy loss caused by impaired uterine perfusion. PMID- 12907108 TI - Sexual activities and attitudes of women with vulvar dysesthesia. AB - OBJECTIVE: To assess the relationship between sexual activities and attitudes of women with and without vulvar dysesthesia. METHODS: Women with vulvar dysesthesia, 18-60 years old, and ethnically matched women without this disorder were enrolled in this cross-sectional study, completed a 27-page questionnaire, and had a physical examination. We compared sexual activities and attitudes between patients and controls using univariate and multivariable analyses. RESULTS: Between January 21, 2001, and December 12, 2002, we enrolled 63 women with vulvar dysesthesia and 62 controls who reported having a current sexual partner. Women with vulvar dysesthesia were less likely than controls to have had, during the previous month, intercourse (66.7% versus 83.9%, P =.03) and orgasms (57.6% versus 78.7%, P =.02), although the association with intercourse was no longer statistically significant after controlling for age, education, and smoking status (P =.07). Of those having had intercourse in the past month, the frequency of this activity was less among patients (3.0 +/- 2.7 versus 6.2 +/- 4.5 episodes, P <.001). Frequencies of orgasms, fellatio, cunnilingus, and masturbation did not significantly differ between patients and controls. Patients rated sex less important and rated themselves more negatively as sexual people than did controls (P <.001). CONCLUSION: Women with vulvar dysesthesia are similar to those without in many sexual activities but are less likely to have had an orgasm in the past month or to have had intercourse at the same frequency as controls. Although some attitudes were similar, sexual interest and self concept were decreased among those with this disorder. PMID- 12907109 TI - A comparison of the effects of epidural and meperidine analgesia during labor on fetal heart rate. AB - OBJECTIVE: To estimate the effects of initiation of epidural analgesia on fetal heart rate (FHR) patterns compared with intravenous meperidine analgesia. METHODS: Fetal heart rate patterns in 200 nulliparous women with term pregnancies randomized to epidural analgesia with 0.25% bupivacaine were compared with those of 156 similar women given intravenous meperidine. Fetal heart rate patterns occurring within 40 minutes of initiation of labor analgesia were retrospectively read by three maternal-fetal medicine specialists who were blind to clinical events, including type of labor analgesia. RESULTS: Meperidine, compared with epidural analgesia, was associated with statistically significantly less beat-to beat variability (absent or less than 5 beats per minute) of the FHR (30% versus 7% of fetuses, P <.001) in the first 40 minutes after initiation of analgesia, as well as with fewer FHR accelerations (88% versus 62% of fetuses, P <.001). Neither the incidence of FHR decelerations nor the type of deceleration were significantly different between methods of labor analgesia. Specifically, 41% of women given meperidine exhibited FHR decelerations within 40 minutes, compared with 34% given epidural analgesia (P =.353). CONCLUSION: Epidural analgesia does not have deleterious effects on FHR. PMID- 12907110 TI - Gestational age-dependent reference values for pH in umbilical cord arterial blood at term. AB - OBJECTIVE: Despite a change of fetal physiologic and biochemical functions with gestational age, stationary umbilical cord acid-base reference values are used as measures of obstetric care quality. The aim of this study was to assess the risk of low Apgar score relative to low pH when using stationary versus gestational age-adjusted reference values. METHODS: Umbilical artery pH was determined in 24,390 term singleton vaginal deliveries with a 5-minute Apgar score of 9 or greater. Low pH was defined as less than 7.10 as a stationary cutoff value, whereas pH less than (mean - 2 standard deviations [SDs]) was used when gestational age was considered. The 5-minute Apgar score less than 7 at low pH was studied in 44,978 term deliveries. RESULTS: A significant negative correlation was found between gestational age and umbilical artery pH. The odds ratio (OR) for pH less than 7.10 was continuously increasing, from 0.6 at 37 weeks to 1.5 at 42 weeks. In contrast, the OR for pH less than (mean - 2 SDs) was steady until 42 weeks, whereafter it increased (OR 1.24; 95% confidence interval 1.05, 1.47). A linear decrease of the association between Apgar score less than 7 and pH less than 7.10 with increasing pregnancy duration was found (P =.097), but for pH less than (mean - 2 SDs) there was no such association. CONCLUSION: We found a physiologic linear decline of umbilical artery pH with gestational age at term. Gestational age-adjusted umbilical cord artery pH reference values result in fewer diagnoses of cord acidemia than a stationary cutoff of pH less than 7.10. PMID- 12907111 TI - Decline in fish consumption among pregnant women after a national mercury advisory. AB - OBJECTIVE: A well-publicized January 2001 federal advisory recommended that pregnant women limit consumption of certain fish because of concerns about mercury contamination. We endeavored to estimate the extent to which pregnant women changed fish consumption habits after dissemination of this national advisory. METHODS: We performed interrupted time series analysis of data from a cohort of pregnant women (2235 who completed at least one dietary questionnaire) visiting obstetric offices in a multispecialty group practice in eastern Massachusetts, surveyed before the advisory from April 1999 through December 2000 and after the advisory from April 2001 through February 2002. Main outcome measures were consumption of total fish and of four fish types: canned tuna, dark meat fish, shellfish, and white meat fish. Subjects reported fish consumption on semiquantitative food frequency questionnaires administered at each trimester of pregnancy. RESULTS: We observed diminished consumption of dark meat fish, canned tuna, and white meat fish after the national mercury advisory. These decreases resulted in a reduction in total fish consumption of approximately 1.4 servings per month (95% confidence interval 0.7, 2.0) from December 2000 to April 2001, with ongoing declines through the end of the study period. There was no change in shellfish intake. CONCLUSION: After dissemination of federal recommendations, pregnant women in this cohort reported reduced consumption of fish, including tuna, dark meat fish, and white meat fish. Because these fish may confer nutritional benefits to mother and infant, public health implications of these changes remain unclear. PMID- 12907112 TI - Antenatal corticosteroids: are incomplete courses beneficial? AB - OBJECTIVE: To assess the effectiveness of an incomplete course of antenatal corticosteroids on perinatal morbidity and mortality. METHOSD: We studied consecutive neonates born at 23-34 weeks' gestation between January 1, 1998, and July 31, 2002. The study population was divided into a group exposed to one 12-mg dose of betamethasone before delivery and a non-corticosteroid-exposed group, which were compared for various perinatal outcome variables. The Student t test, chi(2), Fisher exact tests, and logistic regression were used for analysis. RESULTS: One hundred twenty-five neonates (55.6%) were exposed to one 12-mg dose of betamethasone before delivery, whereas 104 neonates (45.4%) did not receive antenatal corticosteroids. Mean gestational age at delivery (28.4 +/- 3.1 weeks versus 29.7 +/- 3.5 weeks, P =.002) and birth weight (1189 +/- 409 g versus 1289 +/- 441 g, P =.07) were lower among the steroid-exposed group. Adjusting for gestational age, one dose of betamethasone was associated with significant reduction in the need for vasopressors (odds ratio [OR] 0.35; 95% confidence interval [CI] 0.14, 0.85; P =.02), the rate of intraventricular hemorrhage (OR 0.42; 95%CI 0.19, 0.92; P =.03), and neonatal death (OR 0.31; 95% CI 0.11, 0.86; P =.02). There were no differences between groups in the rate of clinical and histological chorioamnionitis, low Apgar scores, postnatal surfactant exposure, respiratory distress syndrome, bronchopulmonary dysplasia, necrotizing enterocolitis, patent ductus arteriosus, retinopathy of prematurity, and neonatal sepsis. CONCLUSION: An incomplete course of antenatal corticosteroids is associated with reduction in the need for vasopressors, the rate of intraventricular hemorrhage, and neonatal death in preterm neonates. PMID- 12907113 TI - Statistics, not memories: what was the standard of care for administering antenatal steroids to women in preterm labor between 1985 and 2000? AB - We determined the frequency of antenatal corticosteroid use for mothers with threatened premature delivery in 1985, 1990, 1995, and 2000. We next compared published data to the surveyed recollections of 302 obstetricians who were practicing during these years. Two points emerged. First, published reports reveal that the use of antenatal corticosteroids increased steadily, from 8% in 1985 to 20% in 1990, 52% in 1995, and 75% in 2000 (P <.001). Second, "expert" opinions derived from the recollections of practicing obstetricians consistently overestimated the actual use of antenatal corticosteroids during the year in question-31% versus 8% for 1985, 56% versus 20% for 1990, 78% versus 52% for 1995, and 92% versus 72% for 2000 (all Ps <.001). The use of antenatal corticosteroids by obstetricians in the past 15 years reveals a phenomenon that is widely recognized elsewhere-retrospective memories are often wrong, and when they are wrong they are not randomly wrong. Rather, recollections are systematically skewed toward an outcome that, in hindsight, is considered desirable (the "Monday morning quarterback" phenomenon). We offer a simple proposal. In determining the "standard of medical care," the legal system should rely on statistical data about doctors' performance rather than the recollections of experts about doctors' performance. The fallible memories of isolated experts are a crude second-best, far inferior to the data that they approximate. Widespread adoption of this view by professional physician organizations would dramatically increase the rationality of expert testimony in medical malpractice tort law. PMID- 12907114 TI - Recurrence of mild malformations and dysplasias. AB - OBJECTIVE: To estimate whether women delivering infants with mild malformations are at increased risk to have a subsequent infant with a mild malformation. METHODS: Both severe and mild malformations detected at birth were cataloged prospectively for 33,701 women with two consecutive singleton births of infants weighing 500 g or more at a tertiary care hospital. Records from a total of 67,402 infants were analyzed from January 1, 1988, through December 31, 2000. Mild malformations and dysplasias were defined to include skin lesions (eg, cafe au lait spots, nevi, and hemangiomas), extra nipples, and abnormalities involving digits. Pearson and McNemar chi(2) statistics and analysis of variance were used for statistical analysis. Estimation of recurrence risks was accomplished using standard methods for rates and proportions. RESULTS: Of the study women, 2.7% delivered infants with mild malformations in their index pregnancy. Mild malformations recurred in 7% of women whose index infant had a mild malformation (2.7% versus 7%, P <.001). Mild malformations involving the skin or digits also significantly increased in the next delivery (2% versus 5%, P <.001; 0.5% versus 8%, P <.001; recurrence of skin and digit anomalies, respectively). CONCLUSION: Women delivering infants with mild malformations involving the skin and digits of the infant are at increased risk for recurrence during their next pregnancy. PMID- 12907115 TI - Prenatal bladder drainage in the management of fetal lower urinary tract obstruction: a systematic review and meta-analysis. AB - OBJECTIVE: To estimate the effect of prenatal bladder drainage on perinatal survival in fetuses with lower urinary tract obstruction. DATA SOURCES: Relevant articles were identified by searching the databases MEDLINE (1966-2002), EMBASE (1988-2002), and the Cochrane library (2000;4). METHODS OF STUDY SELECTION: Studies were selected if the effect of prenatal bladder drainage (vesicocentesis, vesicoamniotic shunt, or open fetal bladder surgery) on perinatal survival was reported in fetuses with ultrasonic evidence of lower urinary tract obstruction. Study selection, quality assessment, and data abstraction were performed independently and in duplicate. TABULATION, INTEGRATION, AND RESULTS: Sixteen observational studies that included nine case series (147 fetuses) and seven controlled series (195 fetuses) were identified. Study characteristics and quality were recorded for each study. Data on the effect of bladder drainage on perinatal survival were abstracted. Where controlled data were available, 2 x 2 tables were generated to compare the effects of bladder drainage versus no bladder drainage on perinatal survival. Pooled odds ratios (ORs) were used as summary measures of effect, and the results were stratified according to predicted fetal prognoses (based on ultrasound features and fetal urinary electrolytes). Among controlled studies, bladder drainage appeared to improve perinatal survival relative to no drainage (OR 2.5; 95% confidence interval [CI] 1.1, 5.9; P =.03). However, this observation was largely because among the subgroup of fetuses with a poor prognosis there was a marked improvement (OR 8.1; 95% CI 1.2, 52.9; P =.03). Improved perinatal outcome was also suggested in those fetuses considered to have a good prognosis (OR 2.8; 95% CI 0.7, 10.8; P =.13). CONCLUSION: There is a lack of high quality evidence to reliably inform clinical practice regarding prenatal bladder drainage in fetuses with ultrasonic evidence of lower urinary tract obstruction. The limited available evidence suggests that prenatal bladder drainage may improve perinatal survival in these fetuses, particularly those with poor predicted prognoses. Further research in the form of a multicenter randomized controlled trial is required to assess the short- and long-term effects of this intervention. PMID- 12907116 TI - Evidence for asymmetric distribution of sciatic nerve endometriosis. AB - OBJECTIVE: To investigate if a lateral asymmetry exists in the distribution of endometriotic lesions of the sciatic nerve. DATA SOURCES: All articles on sciatic nerve endometriosis identified by MEDLINE and EMBASE database searches were retrieved, and additional reports were collected by systematically reviewing all references. Monographs on endometriosis published in the last 15 years were consulted. METHODS OF STUDY SELECTION: We considered articles in which the presence of an endometriotic lesion of the sciatic nerve and the affected side were assessed. We also included reports lacking histological examination of sciatic nerve specimens but with a surgical diagnosis of pelvic endometriosis. Two authors abstracted data independently on standardized forms. The number of women and the side of the lesion were obtained from individual studies, and the combined frequency of left- and right-side sciatic nerve endometriosis in published reports was computed. TABULATION, INTEGRATION, AND RESULTS: Thirty-two reports including 63 subjects were selected. Endometriosis of the sciatic nerve was on the right side in 41 patients, on the left in 20, and bilateral in two. Considering only patients with unilateral sciatic nerve endometriosis, the observed proportion of right-side lesions (41 of 61 [67.2%]; 95% confidence interval 54.0%, 78.7%) significantly differed from the expected proportion of 50% (chi(2)(1) 7.23, P =.007). Among the 16 cases of histological demonstration of endometriosis infiltrating sciatic nerve roots or fibers, ten had it on the right side (62.5%) and six on the left. Twenty-six of the 38 subjects (68.4%) with surgical demonstration of pelvic endometriosis but without histopathologic evidence of direct sciatic nerve involvement were affected by right cyclic sciatica. CONCLUSION: The finding that two thirds of patients with sciatic nerve endometriosis had right-side lesions constitutes further evidence against the coelomic metaplasia theory. The interposition of the sigmoid colon between the regurgitated endometrial cells implanted on the left posterolateral pelvic peritoneum seems to protect the left lumbosacral plexus and sciatic nerve. PMID- 12907117 TI - Simulation laboratories for training in obstetrics and gynecology. AB - Simulations have been used by the military, airline industry, and our colleagues in other medical specialties to educate, evaluate, and prepare for rare but life threatening scenarios. Work hour limits for residents in obstetrics and gynecology and decreased patient availability for teaching of students and residents require us to think creatively and practically on how to optimize their education. Medical simulations may address scenarios in clinical practice that are considered important to know or understand. Simulations can take many forms, including computer programs, models or mannequins, virtual reality data immersion caves, and a combination of formats. The purpose of this commentary is to call attention to a potential role for medical simulation in obstetrics and gynecology. We briefly describe an example of how simulation may be incorporated into obstetric and gynecologic residency training. It is our contention that educators in obstetrics and gynecology should be aware of the potential for simulation in education. We hope this commentary will stimulate interest in the field, lead to validation studies, and improve training in and the practice of obstetrics and gynecology. PMID- 12907118 TI - What matters matter? P values, H values, leadership, and us. AB - Framed by the question "What matters matter?," this essay considers today's physicians' need for leadership, the principled road they embarked on, and the reasons to continue. Taken as a whole, the vast problems of health care seem unsolvable. Approached in small tangible steps, if not cure, could direction, even inspiration, appear? When people are sick, they look to physicians. By actually caring for patients, physicians have earned trust and learned, scientifically and artfully, about life in ways others cannot. Meaningful patient centered care occurs at the junction of logical science and tenuous human needs: "p values" and "h values." Along with the privileged understanding gained from patients comes the responsibility to stand publicly for the rights of all patients to private moments. Standing up in these ways can never be easy; then again, it never was. It is the continued journey toward historic ideals. It is an imprecise place of struggle, where caring "leadership" has always been most needed, fulfilling and truly defining physicians. Despite today's seemingly insurmountable obstacles, in this place each physician can find ways to reenergize around what matters matter. PMID- 12907119 TI - Evaluation and management of women with endometriosis. AB - Endometriosis is a condition that often leads to a variety of symptoms that range from pain complaints to infertility. Endometriosis is also found in women who are asymptomatic. The diagnosis of endometriosis can be made clinically with reliability similar to that of one made surgically. Medical treatment options are effective, as are surgical treatment options. Complications associated with surgery, however, push the balance in favor of medical therapy whenever possible. Based on the body of evidence available at present, women with endometriosis related complaints should be treated with a first-line medical therapy. If that fails, a second-line medical therapy is warranted under most conditions. Laparoscopic surgery should be reserved for patients in whom second-line medical therapy has failed or is contraindicated by desire to conceive immediately or as soon as possible. PMID- 12907120 TI - Incorporation of inhibin-A in second-trimester screening for Down syndrome. PMID- 12907122 TI - Prevention of early-onset group B streptococcal disease in newborns. PMID- 12907124 TI - ACOG Practice Bulletin: clinical management guidelines for obstetrician gynecologists. Number 45, August 2003. Cervical cytology screening (replaces committee opinion 152, March 1995). AB - Although cervical cancer was the leading cause of cancer death in American women as recently as the 1930s, both the incidence and mortality from cervical cancer have decreased by almost one half since the early 1970s, largely as a result of widespread screening with the Pap test. However, the annual incidence rate has remained at approximately 8 cases per 100,000 women over the past few years. New technology for performing cervical cytology is evolving rapidly as are recommendations for classifying and interpreting the results. The purpose of this document is to provide a review of the best available evidence on screening for cervical cancer. Specific equipment and techniques for performing cervical cytology and interpretation of the results are discussed elsewhere. PMID- 12907126 TI - ACOG Committee Opinion Number 284, August 2003: Nonobstetric surgery in pregnancy. AB - Although there are no data to support specific recommendations regarding nonobstetric surgery and anesthesia in pregnancy, it is important for nonobstetric physicians to obtain obstetric consultation before performing nonobstetric surgery. The decision to use fetal monitoring should be individualized, and each case warrants a team approach for the optimal safety of the woman and her baby. PMID- 12907127 TI - ACOG Committee Opinion. Number 285, August 2003: Induced abortion and breast cancer risk. AB - The purpose of this Committee Opinion is to provide a review of recent studies regarding the potential relationship between induced abortion and subsequent breast cancer and to discuss methodologic challenges in this field of study. The American College of Obstetricians and Gynecologists' Committee on Gynecologic Practice concludes that early studies of the relationship between prior induced abortion and breast cancer risk have been inconsistent and are difficult to interpret because of methodologic considerations. More rigorous recent studies argue against a causal relationship between induced abortion and a subsequent increase in breast cancer risk. PMID- 12907129 TI - Effects of fatty acids on the growth of Caco-2 cells. AB - Epidemiological studies suggest that polyunsaturated fatty acids may protect against colorectal neoplasia. In order to explore this observation, cell proliferation and viability, lipid composition, membrane fluidity, and lipid peroxidation were measured in Caco-2 cells after 48h incubation with various fatty acids. Saturated and monounsaturated fatty acids incorporated less well in the membranes than polyunsaturated fatty acids (PUFAs). All of the PUFAs tested had an inhibitory effect on cell proliferation/viability whereas the saturated and monounsaturated fatty acids did not. Addition of palmitic acid had no significant effect on membrane fluidity whereas unsaturated fatty acids increased membrane fluidity in a dose-dependent manner. PUFAs strongly increased tumor cell lipid peroxidation in a dose-dependent manner. Saturated and monounsaturated fatty acids increased lipid peroxidation in this cell line only at high concentration. Preincubation of Caco-2 cells with vitamin E prevented the inhibition of proliferation/viability, the elevation of the MDA concentration and the increased membrane fluidity induced by PUFAs. Our data indicate that PUFAs are potent inhibitors of the growth of colon cancer cells in vitro. PMID- 12907130 TI - Mechanism of garlic (Allium sativum) induced reduction of hypertension in 2K-1C rats: a possible mediation of Na/H exchanger isoform-1. AB - Garlic causes reduction in blood pressure (BP), however the role of Na/H exchanger (NHE) which mediates hypertension and related tissue-damage is poorly understood. In this study the effect of an established dose of raw garlic extract was investigated on the expression of NHE-1 and -3 and sodium pump activity in a 2K-1C model of hypertension in rats. 2K-1C animals showed high BP, increased serum concentration of PGE2 and TxB2, hypertrophy of the unclipped kidneys, but not in the clipped kidneys In addition, NHE-1 and NHE-3 isoforms were increased in both the 2K-1C kidneys, whereas alpha-actin was increased in the clipped but not in unclipped kidneys. Sodium pump activity was decreased in the clipped kidneys, but remained unchanged in the unclipped kidneys. Garlic treatment reduced the induction of NHE-1 only in the unclipped 2K-1C kidneys, whereas garlic treatment increased the sodium pump activity in both the 2K-1C kidneys. These findings demonstrate that the antihypertensive action of garlic is associated with a reversal of NHE-1 induction in the unclipped kidneys. Induction of NHE isoforms together with a reduced sodium pump activity might cause necrosis in the 2K-1C clipped kidneys due to cellular retention of Na+. On the other hand, activation of sodium pump by garlic extract in the kidneys should reduce intracellular Na+ concentration and normalize BP. These findings signify the use of garlic in the treatment of hypertension. PMID- 12907131 TI - Effects of captopril and losartan on lipid peroxidation, protein oxidation and nitric oxide release in diabetic rat kidney. AB - Increased oxidative stress has an important role in the pathogenesis of diabetic nephropathy. The aim of this study was to evaluate the effects of renin anigiotensin system blockage, either by angiotensin-converting enzyme inhibition or angiotensin receptor blockage, on oxidative stress and nitric oxide release in diabetic rat kidneys. After induction of diabetes, six rats were given captopril, six rats were given losartan, and six rats served as diabetic controls. Six healthy rats were also included. At the end of an 8-week period nitric oxide release, lipid peroxidation and protein oxidation were measured in kidney cortices, and urinary albumin excretion (UAE) was determined in 24-h urine samples. Losartan- and captopril-treated diabetic rats had lower levels of UAE than diabetic controls. Diabetic rats had higher levels of lipid peroxidation and protein oxidation compared to healthy rats. NO release was significantly lower in diabetic groups than healthy controls. UAE levels showed a positive correlation with lipid peroxidation and a negative correlation with NO release. Inhibition of lipid peroxidation could be one of the protective mechanisms of renin-angiotensin axis inhibition in diabetic kidney tissues. PMID- 12907132 TI - Macrolide antibiotics inhibit prostaglandin E2 synthesis and mRNA expression of prostaglandin synthetic enzymes in human leukocytes. AB - We investigated the action of macrolide antibiotics, which are considered to have anti-inflammatory activity, on lipopolysaccharide (LPS)-stimulated prostaglandin (PG) E2 synthesis and the expression of mRNAs for cytosolic phospholipase A2 (cPLA2), cyclooxygenase (COX)-1, and COX-2 in human leukocytes. The production of LPS-stimulated PGE2 was significantly increased in peripheral polymorphonuclear leukocytes (PMNLs) and in mononuclear leukocytes (MNLs). Amounts of mRNAs for COX 2 and cPLA2, but not for COX-1, were enhanced by LPS in PMNLs and MNLs. The LPS enhanced PGE2 synthesis and the expression of cPLA2 and COX-2 mRNAs were inhibited by clarithromycin, azithromycin and dexamethasone in PMNLs and MNLs. The mRNA expression of COX-1 in PMNLs was decreased by clarithromycin and azithromycin. Macrolide antibiotics inhibited PGE2 synthesis in human leukocytes by suppressing cPLA2, COX-1, and COX-2 mRNA expression. These data indicate one mechanism of macrolide anti-inflammatory activity. PMID- 12907133 TI - Increased risk of postpartum depressive symptoms is associated with slower normalization after pregnancy of the functional docosahexaenoic acid status. AB - Observational studies suggest an association between a low docosahexaenoic acid (DHA, 22:6n-3) status after pregnancy and the occurrence of postpartum depression. However, a comparison of the actual biochemical plasma DHA status among women with and without postpartum depression has not been reported yet. The contents of DHA and of its status indicator n-6 docosapentaenoic acid (n-6DPA, 22:5n-6) were measured in the plasma phospholipids of 112 women at delivery and 32 weeks postpartum. At this latter time point, the Edinburgh Postnatal Depression Scale (EPDS) questionnaire was completed to measure postpartum depression retrospectively. The EPDS cutoff score of 10 was used to define 'possibly depressed' (EPDS score > or =10) and non-depressed women (EPDS score <10). Odds ratios (OR) were calculated using a multiple logistic regression analysis with the EPDS cutoff score as dependent and fatty acid concentrations and ratio's as explanatory variables, while controlling for different covariables. The results demonstrated that the postpartum increase of the functional DHA status, expressed as the ratio DHA/n-6DPA, was significantly lower in the 'possibly depressed' group compared to the non-depressed group (2.34+/ 5.56 versus 4.86+/-5.41, respectively; OR=0.88, P=0.03). Lactating women were not more predisposed than non-lactating women were to develop depressive symptoms. From this observation it seems that the availability of DHA in the postpartum period is less in women developing depressive symptoms. Although further studies are needed for confirmation, increasing the dietary DHA intake during pregnancy and postpartum, seems prudent. PMID- 12907134 TI - The fluidity of the serum phospholipids of Fulani pastoralists consuming a high fat diet. AB - The semi-nomadic cattle Fulani of northern Nigeria consume a diet rich in saturated fatty acids. Since the quality of an individual's dietary fat can influence the fatty acid composition of their membrane phospholipids (PL), we investigated the effect consumption of relatively large amounts of saturated fat might have on the fatty acid composition and fluidity of the serum PL of the Fulani. We obtained blood serum from 112 Fulani pastoralists (38 males, 74 females) 15-77 years of age and determined the serum fatty acid composition of the total PL fraction of each specimen. Our results indicate that the PL of the Fulani were enriched for saturated fatty acids. The unsaturated/saturated fatty acid ratio was 1.02 for the Fulani PL compared to 1.22-2.08 for seven other reference groups drawn from published reports. In addition, the mean melting point (MMP) of the fatty acyl chains of the serum PL of the Fulani was considerably higher than that of the reference populations (MMP, 30.6 degrees C versus 21.3-26.1 degrees C, respectively). The double bond index (DBI) of the serum PL of the Fulani was much lower than that of the PL of the groups against which comparisons were made (DBI, 0.98 versus 1.24-1.43, respectively). Since serum PL and tissue PL are in dynamic equilibrium, these findings suggest that the tissue PL of the Fulani we studied has considerably less fluid character than those of other populations. Since a variety of membrane functions depend on the fluid property of the acyl chains of their constituent PL, it is conceivable that certain critical membrane-dependent systems, including receptor-ligand interactions, solute transport, enzyme activity and lateral movement of macromolecules, are affected in the Fulani. PMID- 12907135 TI - Potential role of dietary omega-3 essential fatty acids on some oxidant/antioxidant parameters in rats' corpus striatum. AB - Omega-3 (omega-3) is an essential fatty acid (EFA) found in large amounts in fish oil. It contains eicosapentaenoic acid and docosahexaenoic acid (DHA). DHA is one of the building structures of membrane phospholipids of brain and necessary for continuity of neuronal functions. Evidences support the hypothesis that schizophrenia may be the result of increased reactive oxygen species mediated neuronal injury. Recent reports also suggest the protective effect of omega-3 EFA against neuropsychiatric disorders including schizophrenia. This study proposed to assess the changes in antioxidant enzyme and oxidant parameters in the corpus striatum (CS) of rats fed with omega-3 EFA diet (0.4g/kg/day) for 30 days. Eight control rats and nine rats fed with omega-3 were decapitated under ether anesthesia, and CS was removed immediately. Thiobarbituric acid-reactive substances (TBARS) and nitric oxide (NO) levels as well as total superoxide dismutase (t-SOD) and xanthine oxidase (XO) enzyme activities in the CS were measured. Rats treated with omega-3 EFA had significantly lower values of TBARS (P<0.001), NO (P<0.002) and XO (P<0.005) whereas higher values of t-SOD enzyme activity (P<0.002) than the control rats. These results indicate that omega-3 EFA rich fish oil diet reduces some oxidant parameters in CS. This may be revealed by means of reduced CS TBARS levels as an end product of lipid peroxidation of membranes in treated rats. Additionally, reduced XO activity and NO levels may support this notion. On the other hand, although the mechanism is not clear, omega-3 EFA may indirectly enhance the activity of antioxidant enzyme t-SOD. Taken together, this preliminary animal study provides strong support for a therapeutic effect of omega-3 EFA supplemented to classical neuroleptic regimen in the treatment of schizophrenic symptoms and tardive dyskinesia. PMID- 12907136 TI - Effects of taurolidine and octreotide on tumor growth and lipid peroxidation after staging-laparoscopy in ductal pancreatic cancer. AB - Irrigation with taurolidine after laparoscopy decreases tumor growth in colon carcinoma. In pancreatic cancer subcutaneous therapy with octreotide decreases oxidative stress and carcinogenesis as well. However, it is still unclear, whether irrigation with taurolidine or octreotide after laparoscopic pancreatic biopsy reduces tumor growth in pancreatic cancer as well. In 60 Syrian hamsters ductal pancreatic adenocarcinoma was induced by weekly injection of 10mg/kg body weight N-nitrosobis-2-oxopropylamine s.c. for 10 weeks. In week 16 laparoscopic pancreatic biopsy by use of carbon dioxide was performed (gr. 1, n = 20) with subsequent laparoscopic irrigation with taurolidine (gr. 2, n = 20) or octreotide (gr. 3, n = 20). In week 25 hamsters were sacrificed. Our results show that macroscopic visible primary tumors were found in only one animal of the taurolidine group (5.9%), compared to 42.1% in the saline and 62.5% in the octreotide group (P<0.05). Carcinomas were smaller after saline (6+/-23 mm(2)) than after octreotide irrigation (70+/-120 mm(2), P<0.05). In conclusion this study showed that laparoscopic irrigation with taurolidine after pancreatic biopsy inhibited tumor growth in ductal pancreatic adenocarcinoma. PMID- 12907137 TI - Effects of orally administered OP-1206 alpha-CD with loxoprofen-Na on walking dysfunction in the rat neuropathic intermittent claudication model. AB - An orally active prostaglandin E1 analogue, OP-1206 alpha-CD improves walking dysfunction in the rat spinal stenosis model. Loxoprofen-Na, a non-steroidal anti inflammatory drug, is used to relieve chronic pain in patients with lumbar spinal canal stenosis. To determine whether the OP-1206 alpha-CD in combination with loxoprofen-Na could induce a greater therapeutical effect on walking dysfunction and spinal cord blood flow (SCBF) than OP-1206 alpha-CD treatment alone after chronic spinal stenosis in the rat. Spinal stenosis was induced by placing two pieces of silicon rubber strips in the lumbar (L4 and L6) epidural space of rats. After surgery, walking function was measured using a treadmill apparatus and SCBF was measured using a laser-Doppler flow meter. Drugs were administered orally twice a day for 11 days from the day 3 post-surgery. OP-1206 alpha-CD elicited a significant improvement of walking dysfunction on days 7 and 14 post-surgery and significantly increased spinal cord blood flow on day 15, whereas walking dysfunction and SCBF of rats treated with loxoprofen-Na alone remained unchanged. Combined treatment of OP-1206 alpha-CD with loxoprofen-Na did not provide additive therapeutical effect. These results suggest that a significant improvement seen after OP-1206 alpha-CD treatment is primarily mediated by improvement of the local spinal cord blood flow. This effect is not ameliorated or potentiated by a combined treatment with loxoprofen-Na. PMID- 12907138 TI - Levels of expression of lipoxygenases and cyclooxygenase-2 in human breast cancer. AB - Lipoxygenases and cyclooxygenase are key mediators of arachidonic acid metabolism. The eicosanoids metabolites from these oxygynases have been shown to regulate the growth and death of cancer cells. This study determined the level of expression of 5-, 12-, 15-lipoxygenase and cyclooxygenase-2 expression in a cohort of breast cancer patients and their correlation with clinical outcomes. Compared with normal breast tissues, tumour tissues exhibited a significantly higher levels of 12-lipoxygenase and cyclooxygenase-2 (P<0.05), and significantly lower level of 15-lipoxygenase (P=0.05). Lobular carcinomas had a higher level of cyclooxygenase-2 and lower level of 15-lipoxygenase than ductal carcinomas. The lowest level of 15-lipoxygenase was seen in TNM3 and TNM4 tumours and from patients who died of breast cancer. Levels of 12- and 5-lipoxygenases were also particularly high in tumours from patients who died of breast cancer. This study shows that human breast tumours aberrantly express lipoxygenases and cyclooxygenase-2 and that decreased level of 15-lipoxygenase and raised level of cyclooxygenase-2 and 12-lipoxygenase has prognostic value in patients with breast cancer. PMID- 12907139 TI - Conversion of alpha-linolenic acid to palmitic, palmitoleic, stearic and oleic acids in men and women. AB - The purpose of this study was to determine whether adult humans can recycle carbon from alpha-linolenic acid (18:3n-3) into saturated (SFA) and monounsaturated (MUFA) fatty acids. Six men and six women consumed 700 mg [U-13C] 18:3n-3. Blood was collected over 21 days and breath over 24h. [13C]-labelled SFA and MUFA were detected in plasma phosphatidylcholine (PC) and triacylglycerol (TAG). Total labelled fatty acid incorporation into SFA and MUFA was five- and 25 fold greater in PC than TAG in men and women, respectively. [13C]-16:0 was the major labelled fatty acid in both fractions. Total [13C] incorporation into SFA and MUFA was 20% greater in men than women, and related positively (r(2) = 0.35, P<0.05) to the fractional recovery of labelled 18:3n-3 as 13CO2 on breath. These results suggest that the extent of partitioning towards beta-oxidation and carbon recycling may regulate the availability of 18:3n-3 for conversion to longer-chain fatty acids. PMID- 12907140 TI - American Society of Gene Therapy (ASGT) ad hoc subcommittee on retroviral mediated gene transfer to hematopoietic stem cells. PMID- 12907141 TI - AAV-Mediated gene transfer slows photoreceptor loss in the RCS rat model of retinitis pigmentosa. AB - In the Royal College of Surgeons (RCS) rat, the retinal pigment epithelium (RPE) cannot phagocytose the outer segment discs that are continually shed from photoreceptors. The resulting accumulation of debris in the subretinal space leads to a progressive loss of photoreceptors. The defect results from a mutation in the Mertk gene, which is normally expressed in the RPE. Mertk is a receptor tyrosine kinase, involved in the binding of photoreceptor debris. Mutations in MERTK have also been described in patients with retinitis pigmentosa (RP). Here we demonstrate that subretinal injection of recombinant adeno-associated virus (AAV) expressing the murine Mertk gene can significantly prolong photoreceptor cell survival in the RCS rat. Electroretinographic analysis of treated eyes showed that functional photoreceptors were still present at 9 weeks, when there is virtually no activity in untreated control eyes. Histological analysis of treated eyes revealed a decrease in the amount of debris in the subretinal space, suggesting that RPE function was restored. Moreover, 9 weeks after treatment the number of photoreceptors was 2.5-fold higher in treated than in control eyes. This study provides strong support for the development of AAV-mediated gene therapy for RP caused by mutations in the MERTK gene. PMID- 12907142 TI - Inhibition of HIV-1 infection by lentiviral vectors expressing Pol III-promoted anti-HIV RNAs. AB - A primary advantage of lentiviral vectors is their ability to pass through the nuclear envelope into the cell nucleus thereby allowing transduction of nondividing cells. Using HIV-based lentiviral vectors, we delivered an anti-CCR5 ribozyme (CCR5RZ), a nucleolar localizing TAR RNA decoy, or Pol III-expressed siRNA genes into cultured and primary cells. The CCR5RZ is driven by the adenoviral VA1 Pol III promoter, while the human U6 snRNA Pol III-transcribed TAR decoy is embedded in a U16 snoRNA (designated U16TAR), and the siRNAs were expressed from the human U6 Pol III promoter. The transduction efficiencies of these vectors ranged from 96-98% in 293 cells to 15-20% in primary PBMCs. A combination of the CCR5RZ and U16TAR decoy in a single vector backbone gave enhanced protection against HIV-1 challenge in a selective survival assay in both primary T cells and CD34(+)-derived monocytes. The lentiviral vector backbone expressed siRNAs also showed potent inhibition of p24 expression in PBMCs challenged with HIV-1. Overall our results demonstrate that the lentiviral-based vectors can efficiently deliver single constructs as well as combinations of Pol III therapeutic expression units into primary hematopoietic cells for anti-HIV gene therapy and hold promise for stem or T-cell-based gene therapy for HIV-1 infection. PMID- 12907143 TI - MDA-7 negatively regulates the beta-catenin and PI3K signaling pathways in breast and lung tumor cells. AB - mda-7 is a novel tumor suppressor with cytokine properties. Adenoviral mda-7 (Ad mda7) induces apoptosis and cell death selectively in tumor cells. The molecular mechanisms underlying the anti-tumor activity of Ad-mda7 in breast and lung cancer lines were investigated. Microarray analyses implicated both the beta catenin and the PI3K signaling pathways. Ad-mda7 treatment increased protein expression from tumor suppressor genes, including E-cadherin, APC, GSK-3beta, and PTEN, and decreased expression of proto-oncogenes involved in beta-catenin and PI3K signaling. Ad-mda7 caused a redistribution of cellular beta-catenin from the nucleus to the plasma membrane, resulting in reduced TCF/LEF transcriptional activity, and upregulated the E-cadherin-beta-catenin adhesion complex in a tumor cell-specific manner. Expression of the PI3K pathway members (p85 PI3K, FAK, ILK 1, Akt, and PLC-gamma) was downregulated and expression of the PI3K antagonist PTEN was increased. Consistent with this result, pharmacological inhibition of PI3K by wortmannin did not abrogate killing by Ad-mda7. Killing of breast cancer cells by Ad-mda7 required both MAPK and MEK1/2 signaling pathways, whereas these pathways were not essential for MDA-7-mediated killing in lung cancer cells. Thus, in breast and lung tumor cells MDA-7 protein expression modulates cell-cell adhesion and intracellular signaling via coordinate regulation of the beta catenin and PI3K pathways. PMID- 12907144 TI - Tumor suppressor MDA-7/IL-24 selectively inhibits vascular smooth muscle cell growth and migration. AB - Abnormalities in smooth muscle cell (SMC) proliferation and differentiation underlie the pathogenesis of proliferative vascular diseases. MDA-7 (HUGO approved symbol IL24) is a unique gene, originally identified as a tumor suppressor and more recently shown to have cytokine activity. MDA-7/IL24 has been implicated in apoptosis and cellular differentiation in tumor cells and in tumor invasion/metastasis in clinical specimens-properties central to SMC remodeling during proliferative vascular diseases. In this study, we evaluated the effects of overexpressing MDA-7/IL24 in various SMC: the apparently "normal" rat PAC1 cell line, primary human coronary artery SMC, and normal rat aortic SMC. We transduced SMC with adenovirus-mda7 (Ad-mda7) or control virus (Ad-Luc) and assessed cell viability, apoptosis, and migration. Ad-mda7 suppressed PAC1 cell growth in a dose-dependent manner while having no effect on normal primary human coronary artery cells or rat aortic SMC, despite strong expression of the MDA-7 transgene in all SMC. Similarly, Ad-mda7 treatment induced apoptosis in PAC1 cells with essentially no effect on normal coronary and rat aortic SMC. Ad-mda7 also inhibited serum-stimulated PAC1 cell migration. Karyotype analysis of PAC1 cells revealed that they exhibit multiple chromosomal aberrations. Importantly, recombinant MDA-7 did not elicit cell death or STAT-3 activation in PAC1 SMC, suggesting that the effects of Ad-mda7 were mediated through an intracellular pathway. These data demonstrate that Ad-mda7 exhibits selectivity in apoptosis induction and growth suppression in an atypical SMC line, raising new questions pertaining to heterogeneity in SMC death susceptibility. PMID- 12907145 TI - RU486-inducible retrovirus-mediated caspase-3 overexpression is cytotoxic to bcl xL-expressing myeloma cells in vitro and in vivo. AB - The antiapoptotic protein bcl-x(L) is upregulated in a variety of solid tumors and in primary hematologic malignancies such as multiple myeloma. Activated caspase-3 cleaves proteins essential for cell survival, including bcl-x(L). To explore the potential of caspase-3 as a cytotoxic and immunostimulatory molecule in the treatment of malignancy, an RU486-inducible caspase-3 retrovirus was constructed, validated, and used to transduce first 3T3 and subsequently murine myeloma B9BM1 cells (creating the cell line B9BM-C3). After induction, apoptotic cell death of 3T3 and B9BM-C3 cells began by 4 h and was complete by 48 h postinduction, while nontransduced cells remained viable. Annexin V staining demonstrated 43, 76, and 98% apoptotic cell death at 12, 18, and 24 h postinduction. Activation of caspase-3 was evident in induced cells and cell death could be inhibited by the addition of a caspase-3-specific inhibitor. Overexpression of the myeloma-associated oncogene FGFR3, which upregulates bcl x(L), delayed but did not prevent caspase-3-mediated killing. B9BM-C3 cells formed tumors after subcutaneous injection in mice. Early treatment with RU486 eradicated tumors; however, rechallenge of treated mice failed to demonstrate evidence of immunoprotection. These results indicate that therapeutic attempts to induce caspase-3 in malignant cells may prove useful in the treatment of bcl-x(L) expressing tumors. PMID- 12907146 TI - Recombinant vaccinia virus expressing IL-2 generates effective anti-tumor responses in an orthotopic murine model of head and neck carcinoma. AB - We evaluated the efficacy of a replication-competent, attenuated recombinant vaccinia virus (rvv) expressing IL-2 as a tumor vaccine in an immunocompetent murine model of head and neck squamous cell carcinoma. We implanted oral tumors by injection of tumor cells (SCC VII/SF) into the floor of the mouth of the syngeneic C3H/HeJ mice. Previous studies with this model suggested the presence of tumor-induced immune suppression. To circumvent the immune suppression we subcutaneously (s.c.) immunized the mice with irradiated, rvv-infected tumor cells prior to or along with intratumoral (i.t.) vaccination. A single s.c. vaccination invoked tumor-specific T cell proliferation and cytotoxicity. Mice treated by this protocol survived longer compared to those treated with i.t. vaccination alone. Tumor growth was significantly inhibited (P < 0.0002) and tumor regression was observed in all mice. The numbers of CD4(+) and CD8(+) lymphocytes as well as macrophages in the tumor beds and in tumor-draining lymph nodes were significantly increased in the mice treated by s.c. plus i.t. vaccination compared to s.c. or i.t. vaccination alone. These results suggested that s.c. vaccination along with i.t. vaccination increased antitumoral immunity and that CD4(+), CD8(+) lymphocytes as well as macrophages may play an important role in this antitumoral immunity. PMID- 12907147 TI - Biodistribution and pharmacokinetics of aerosol and intravenously administered DNA-polyethyleneimine complexes: optimization of pulmonary delivery and retention. AB - This report describes the time-dependent biodistribution of human p53 plasmid delivered in aerosol with polyethyleneimine in mice compared to the distribution of this material following intravenous injection. Area-under-the-curve values for p53 plasmid after inhalation were 2.8-fold greater than values after intravenous administration, despite the fact that the delivered aerosol dose was one-fifth the intravenous dose. After aerosol administration, pulmonary concentrations of p53 plasmid were high and other organs showed amounts not distinguishable from untreated control. High concentrations of p53 plasmid in the lungs remained with negligible reduction for at least 24 h. Shortly after intravenous injection, organs exhibited the following relative levels of exogenously administered p53: liver > spleen > blood > or = lungs > heart > kidney. These results demonstrate effective pulmonary delivery of DNA in complex with PEI by aerosol, without significant systemic dissemination. In contrast, intravenous administration caused a prompt systemic distribution of DNA with a shorter half-life of the administered gene in the lungs. PMID- 12907148 TI - Efficient in vitro and in vivo expression of covalently modified plasmid DNA. AB - The tracking of plasmid DNA (pDNA) movement within cells requires the attachment of labels to the DNA in a manner such that: (a) the pDNA remains intact during the labeling process and (b) the labels remain stably attached to the DNA. Keeping these two criteria in mind, we have recently developed a series of alkylating reagents that facilitate the one-step, covalent attachment of compounds directly onto nucleic acids in a nondestructive manner. Using these DNA alkylating reagents, we have attached a wide range of both fluorescent and nonfluorescent reporter molecules onto pDNAs. We now show that even with the covalent attachment of various marker compounds, the pDNA remains expression competent. The ability to create labeled, expression-competent DNA allows for the simultaneous tracking of both pDNA location and reporter gene expression within living or fixed cells. PMID- 12907149 TI - Identification of human plasma proteins that bind to cationic lipid/DNA complex and analysis of their effects on transfection efficiency: implications for intravenous gene transfer. AB - Interaction of cationic lipid/DNA complex with the plasma is a limiting step for the cationic lipid-mediated intravenous gene transfer and expression process. Most of the plasma components that interact with the complex and inhibit its transfection efficiency are still unknown. In the present work, human plasma proteins and lipoproteins that bind to a cationic lipid/DNA complex were isolated on a sucrose density gradient and identified by 2-D gel electrophoresis. Protein binding did not result in complex dissociation or DNA degradation. The effects of several complex-binding plasma components on the transfection efficiency were studied using lung endothelial cells cultured in vitro. Lipoprotein particles caused a drastic loss of the transfection efficiency of the complex. Surprisingly, fibrinogen was found to activate the transfection process. The roles of these complex-binding plasma components on the complex uptake efficiency were quantitatively assessed using radiolabeled plasmid DNA and qualitatively evaluated using fluorescence microscopy. A good correlation was found between the effects of the complex-binding plasma components on the transfection and on cell uptake efficiencies. In contrast to what was generally believed, our data suggest that disruption of the complex does not occur when it is in contact with the plasma and therefore could not be responsible for the loss of transfection activity. Instead, coating of complexes with plasma components seems to be responsible for reduced uptake by cells, which in turn results in reduced transfection. PMID- 12907150 TI - Identification of an HLA-A*0201-restricted epitopic peptide from human dystrophin: application in duchenne muscular dystrophy gene therapy. AB - Dystrophin-based gene therapy treatments aimed at correcting the Duchenne muscular dystrophy phenotype require stable expression of normal dystrophin (DYST) protein in myocytes without immune responses, which would compromise long term expression. To predict cytotoxic T-cell-mediated responses elicited by transgenes, we used here H-2-negative HLA-A*0201 transgenic mice and identified human DYST epitopes, which elicit HLA-A*0201-restricted cytotoxic T cell activities. Among a series of eight peptides predicted from the human DYST sequence, not shared with the endogenous mouse DYST sequence, four of them were able to bind to HLA-A*0201 molecules and to induce cytotoxic T lymphocyte (CTL) responses. After human DYST DNA transfer in muscle of HLA-A*0201 mice, only the human DYST1281 epitope, located in the spectrin-like repeat 9 domain, induced strong CD8(+) CTL responses. Using the corresponding human DYST1281 peptide/HLA A*0201 tetramer, we detected human DYST1281-specific CD8(+) T cells in peripheral lymphoid organs and blood of HLA-A*0201 mice injected with human DYST DNA. Our results demonstrate that muscle injection with human DYST DNA systematically triggers CTL responses against this HLA-A*0201-restricted human DYST1281 peptide, which is present in long human DYST isoforms. Identification of such immunodominant human DYST epitopes and use of peptide/HLA tetramers will allow the immunomonitoring of CTL responses in HLA-phenotyped Duchenne muscular dystrophy patients undergoing gene therapy. Finally, the knowledge of HLA-A*0201 restricted human DYST peptides will be of importance to test, in mouse models, new immunomodulatory interventions allowing long-term engraftment of human dystrophin. PMID- 12907151 TI - Caveolae-mediated internalization of extracellular HIV-1 tat fusion proteins visualized in real time. AB - The Tat protein from HIV-1, when fused with heterologous proteins or peptides, can traverse cell membranes. This ability has generated great interest due to potential therapeutic applications. However, the relevant cellular pathway and its dynamics have not been elucidated yet. Here we unravel the intracellular fate of exogenously added Tat fused with green fluorescent protein (GFP) in live HeLa and CHO cells, from the early interaction with the plasma membrane up to the long term accumulation in the perinuclear region. We demonstrate that the internalization process of full-length Tat and of heterologous proteins fused to the transduction domain of Tat exploits a caveolar-mediated pathway and is inhibited at 4 degrees C. Remarkably, a slow linear movement toward the nucleus of individual GFP-tagged Tat-filled caveolae with an average velocity of 3 micro m/h was observed. No fluorescence was observed in the nucleus, possibly suggesting that Tat fusion protein unfolding is required for nuclear translocation. In addition, early sensitivity to cytochalasin-D treatment indicates the essential role of the actin cytoskeleton in the displacement of Tat vesicles toward the nucleus. Our results imply that HIV-1 Tat mediates the internalization of protein cargos in a slow and temperature-dependent manner by exploiting the caveolar pathway. PMID- 12907152 TI - Identification of a synovial fibroblast-specific protein transduction domain for delivery of apoptotic agents to hyperplastic synovium. AB - Synovial hyperplasia, resulting in erosion of cartilage and bone, represents one of the major pathologies associated with rheumatoid arthritis. To develop an approach for efficient delivery of proteins or agents to synovium to induce targeted apoptosis of hyperplastic synovial tissue, we have screened an M13 peptide phage display library for synovial-specific transduction peptides. We identified a novel synovial-targeted transduction peptide, HAP-1, which is able to facilitate specific internalization of protein complexes into human and rabbit synovial cells in culture and rabbit synovial lining in vivo. HAP-1 and a non tissue-specific cationic protein transduction domain, PTD-5, were fused to an antimicrobial peptide, (KLAK)(2), to generate two proapoptotic peptides termed DP2 and DP1, respectively. Administration of these peptides was able to induce apoptosis of rabbit and human synovial cells in culture, with DP2 inducing synovial cell-specific apoptosis. Intra-articular injection of DP1 and DP2 into arthritic rabbit joints with synovial hyperplasia induced extensive apoptosis of the hyperplastic synovium, while reducing the leukocytic infiltration and synovitis. These results suggest that proapoptotic peptides and, in particular, DP2 can be clinically useful for treatment of synovial hyperplasia, as well as inflammation. Moreover, the results demonstrate the feasibility of identifying tissue-specific transduction peptides capable of mediating efficient transduction in vivo. PMID- 12907153 TI - Catheter-based intracoronary myocardial adenoviral gene delivery: importance of intraluminal seal and infusion flow rate. AB - Although percutaneous, adenoviral-mediated intracoronary gene delivery to the heart has been demonstrated in some species, consistent and safe methodology is needed before clinical applicability is possible. In this study, we examine the effects of altering intracoronary flow rate and obtaining an adequate seal between the catheter and the coronary lumen on successful cardiac gene delivery and myocardial injury in both piglets and adult rabbits. To study the efficacy of in vivo myocardial gene transfer, we utilized adenoviral vectors containing either the beta(2)-adrenergic receptor or beta-galactosidase. The left circumflex coronary artery of piglets and the right coronary artery of rabbits were catheterized under fluoroscopic guidance and adenovirus solutions were injected using varying flow rates with or without balloon inflation. Successful transgene delivery to the heart was determined approximately 1 week after coronary infusions. Histologic analysis was also performed in all animals to determine the extent of myocardial injury. Our results indicate that efficient and reproducible cardiac transgene expression utilizing intracoronary delivery is dependent upon the infusion flow rate and, in larger animals, requires an intraluminal seal. Excessive flow rate is associated with greater myocardial injury. Thus, conditions can be established and controlled to improve future investigational and clinical application of catheter-based intracoronary myocardial gene therapy. PMID- 12907154 TI - The effect of age on hepatic gene transfer with self-inactivating lentiviral vectors in vivo. AB - It is known that cellular proliferation, by either compensatory regeneration or direct hyperplasia, can augment lentiviral vector transduction into hepatocytes in vivo. For this reason, the present study was designed to determine if adolescent mice (312 weeks of age), which still have relatively proliferating livers, would have differential transduction compared to older (7 weeks of age) mice. Self-inactivating lentiviral vectors containing the human alpha(1) antitrypsin (hAAT) promoter driving the expression of either the bacterial lacZ gene or the hAAT cDNA were generated for these studies. We found that adolescent mice given lentiviral vectors expressing lacZ (50 micro g p24/mouse) via intravenous administration had a significantly higher level of hepatocyte transduction as measured by X-gal staining of liver sections compared to the 7 week-old mice. In addition, serum hAAT levels were nearly 40-fold higher in 312 week-old mice administered lentiviral vectors expressing hAAT (50 micro g p24/mouse) compared to the 7-week-old mice. Moreover, the incorporation of a matrix attachment region from immunoglobulin kappa significantly increased transduction of hepatocytes in vivo. Although there was a small reduction in the circulating levels of hAAT, likely due to an immune response against the transgene product, gene expression was sustained for the duration of the study (30 weeks in total). In conclusion, the present study strongly demonstrates that lentiviral vector transduction efficiency and transgene expression were significantly enhanced in adolescent compared to older mice. PMID- 12907155 TI - HIV vector production mediated by Rev protein transduction. AB - HIV-1-based vectors are promising tools for gene therapy because of their ability to integrate into nondividing cells. Their safety in clinical applications remains a major concern. Recombination events occurring among plasmid constructs during vector production could potentially lead to the generation of replication competent viruses. The safety of HIV-1-based vectors can be improved by removing all regions of the viral genome that are not absolutely required for vector production or function. In this study, we demonstrate that the HIV-1 rev gene is dispensable for the production of HIV-1-based vectors if the vector-producing cells are supplied with purified Rev protein. We compared the efficiency of vector production among Rev, TAT-Rev (Rev fused to the protein transduction domain of the HIV TAT protein), and Rev/Pep-1 (Rev complexed with the carrier peptide Pep-1). Our results showed that 293T cells efficiently internalized TAT Rev and Rev/Pep-1 and high-titer vector preparations were obtained with this approach. Vectors generated by such an approach showed little difference in their efficiencies of transduction of established cell lines and primary cells compared with vectors generated by standard plasmid cotransfection. Eliminating the requirement for the HIV-1 rev gene during vector production should improve the safety of applying HIV vectors in human clinical trials. PMID- 12907156 TI - Development of a sensitive assay for detection of replication-competent recombinant lentivirus in large-scale HIV-based vector preparations. AB - Lentiviral vectors have demonstrated great potential as gene therapy vectors mediating efficient ex vivo and in vivo gene delivery and long-term transgene expression in both dividing and nondividing cells. However, for clinical studies it must be demonstrated that lentiviral vector preparations are safe and not contaminated by replication-competent recombinants related to the parental pathogenic virus. Here we describe a sensitive assay for the detection of replication-competent lentiviruses (RCL) in large-scale preparations of HIV-based lentiviral vectors. This RCL assay for lentiviral vectors is based on the principles used for retroviral vectors, using a highly permissive cell line, C8166-45, for RCL amplification and an appropriate positive control virus to establish the assay sensitivity. The assay is capable of detecting 1 RCL infectious unit in a background of 2.5 x 10(8) transducing units of vector in a single test culture. Statistically representative samples from large-scale lentiviral vector productions were assayed using multiple test cultures for each lot. Overall, a total of 1.4 x 10(10) transducing units of vector from 10 independent 14-liter production lots were screened and no RCL was detected. We propose to implement this assay as a release testing for clinical-grade lentiviral vector preparations intended for gene therapy clinical trials. PMID- 12907157 TI - Femtosecond infrared laser-an efficient and safe in vivo gene delivery system for prolonged expression. AB - The major advantages of "naked DNA gene therapy" are its simplicity and a low or negligible immune response. Gene delivery by DNA electroporation (EP) involves injection of DNA and the application of a brief electric pulse to enhance cellular permeability. Although EP is an efficient gene transduction technique in rodents, it requires much higher voltages (>500 V) in larger animals, and hence, in practice it would be hazardous for human patients, as it would cause serious tissue damage. To overcome the obstacles associated with EP-mediated gene delivery in vivo, we developed a new method of gene transduction that uses laser energy. The femtosecond infrared titanium sapphire laser beam was developed specifically for enhancing in vivo gene delivery without risks of tissue damage. System optimization revealed that injection of 10 micro g naked DNA into the tibial muscle of mice followed by application of the laser beam for 5 s, focused to 2 mm depth upon an area of 95 x 95 micro m(2), resulted in the highest intensity and duration of gene expression with no histological or biochemical evidence of muscle damage. We assessed the potential clinical application of LBGT technology by using it to transfer the murine erythropoietin (mEpo) gene into mice. LBGT-mediated mEpo gene delivery resulted in elevated (>22%) hematocrit levels that were sustained for 8 weeks. Gene expression following LBGT was detected for >100 days. Hence, LBGT is a simple, safe, effective, and reproducible method for therapeutic gene delivery with significant clinical potential. PMID- 12907158 TI - Human beta-crystallins modified by backbone cleavage, deamidation and oxidation are prone to associate. AB - Information about beta-crystallins and their post-translational modifications has been scarce because of difficulties in isolating the individual beta-crystallins. These difficulties arise because the beta-crystallin sequences are highly homologous and because beta-crystallins undergo many age-related modifications that lead to a variety of molecular masses and a range of acidities for each crystallin. In this study, human beta-crystallins were isolated using several steps of chromatography both before and after two-dimensional gel electrophoresis. Many previously unidentified in vivo modifications, including deamidations among all beta-crystallins except betaB3, truncation of betaA3, betaB1 and betaA4, and oxidation of some methionines and tryptophans were located among the isolated beta-crystallins. Many modifications occurred before age 20 with modest increases in modification for beta-crystallins from lenses 20-87 years old. The tendency of the modified beta-crystallins to form non-covalent complexes was evident from their chromatographic behaviour. The presence in these complexes of betaB2-crystallin, the least modified and most soluble of the beta crystallins, points to a possible role for betaB2 in solubilizing the more heavily modified beta-crystallins. The greater solubility of beta-crystallins compared with alpha- and gamma-crystallins in aging lenses may be due to beta crystallin modifications and their non-covalent associations. PMID- 12907159 TI - Isolation of a putative keratocyte activating factor from the corneal stroma. AB - Fetal bovine serum has commonly been used to expand the population of keratocytes in culture. Tissue extracts, however, have also been used to grow other cell types. We prepared a DMEM/F12 extract of corneal stroma and compared the growth and morphology of collagenase-isolated keratocytes cultured in DMEM/F12, or DMEM/F12 containing either stromal extract or fetal bovine serum. Cell proliferation was measured by 3H-thymidine and BrdU incorporation as well as by DNA quantitation. The extract was fractionated by gel filtration. Cell morphology was assessed by phase-contrast microscopy. Culture in both extract and serum stimulated keratocytes to proliferate, but keratocytes cultured in the extract grew more slowly due to a longer cell cycle and to a lower final density because of greater sensitivity to contact inhibition. Keratocytes cultured in serum became fibroblastic while those cultured in extract retained the dendritic morphology of quiescent keratocytes. The stimulating factors in the corneal extract were more sensitive to heat inactivation and of higher molecular weight than the stimulating factors in serum. These results indicate that the mitogenic activity in extract and serum are different and that the phenotypes resulting from growth in serum and extract are also different. Keratocytes cultured at low cell densities in the corneal extract may mimic keratocyte activation, an initial and crucial event for keratocytes during the corneal wound healing process. PMID- 12907160 TI - Integrin alphavbeta5 is not required for the phagocytosis of photoreceptor outer segments by cultured retinal pigment epithelial cells. AB - The phagocytosis of photoreceptor outer segments (OS) by the retinal pigment epithelium (RPE) is a receptor mediated process. A key component of this process is the receptor tyrosine kinase, Mer. RPE cells from the RCS rat, which lacks a functional mer gene, and do not express Mer protein, are able to bind OS, but are unable to ingest them, suggesting that both a binding receptor and an ingestion receptor (Mer) are required for phagocytosis to occur. These rats become blind shortly after birth. To date the binding receptor has not been identified. Recent studies, using an SV40 transformed rat RPE cell line, RPE-J, or cultured human RPE cells, have suggested that the receptor for OS binding is the integrin alphavbeta5. However, the results presented here clearly show that this integrin plays at most a minor role in the phagocytosis of OS by primary cultures of rat RPE cells. OS phagocytosis by normal RPE cells is not affected by a function blocking antibody to alphavbeta5 integrin, nor by the integrin-specific blocking peptide GRGDSP. Additionally, RPE-J cells do not express the Mer receptor protein, which has been shown to be obligatory for OS phagocytosis, or RPE65, a specific marker for RPE cells. We suggest that the RPE-J cell line is not a valid model with which to study the complex process of OS phagocytosis. PMID- 12907161 TI - Identification and cloning of the Na/HCO(3-) cotransporter (NBC) in human corneal endothelium. AB - Fluid secretion by the corneal endothelium is associated with the net flux of HCO(3)(-) from basolateral (stromal) to apical (anterior chamber) sides of the tissue. In this study we asked if Na(+)/HCO(3)(-) cotransporter (NBC-1) protein expression and functional activity are present in freshly isolated human corneal endothelium. Immunoblot analysis using a polyclonal antibody to NBC-1 showed a single band at approximately 130 kDa. Indirect immunofluorescence indicated that NBC-1 is expressed on the basolateral, but not apical side of human corneal endothelium. RT-PCR was used to determine whether the kidney or pancreatic isoform of NBC-1 is expressed. Using the specific primers for pNBC and kNBC isoforms, RT-PCR showed that only pNBC could be detected in human corneal endothelium. The product was cloned and confirmed by sequencing. Full-length NBC 1 was also cloned from human corneal endothelium. This clone (hcNBC) is 100% identical to the longer, more common form of NBC [pNBC; 1079 amino acids (aa); 122 kDa in human heart, pancreas and prostate]. To test for functional activity of NBC-1, freshly isolated endothelium was loaded with the pH sensitive fluorescent dye BCECF and HCO(3)(-) fluxes were measured. HCO(3)(-) fluxes were Na(+)-dependent, electrogenic and H(2)-DIDS sensitive. We conclude that the long isoform of the sodium bicarbonate cotransporter (pNBC-1) is expressed on the basolateral side of fresh human corneal endothelium (hcNBC). The shorter form, kNBC, could not be detected. As in bovine corneal endothelium, hcNBC is instrumental in loading HCO(3)(-) into endothelial cells from the basolateral membrane. PMID- 12907162 TI - Lacrimal gland as the major source of mouse tear factors that are cytotoxic to corneal keratocytes. AB - We have previously shown that mouse tears are cytotoxic to the corneal keratocytes. Since tear components are derived from both lacrimal tissues and ocular surface epithelium, we sought to determine the source of the cytotoxic factors in the mouse tear fluid. Cytotoxicity to keratocytes was assessed by an ex vivo assay using an isolated eye; after treatment with test samples, segmentation and disappearance of stromal nuclei were determined by DAPI nuclear staining. Following biological tissues and fluids were examined either directly or after preincubation at 37 degrees C for 2-15 hr: extraorbital lacrimal gland (ELG), intraorbital lacrimal gland (ILG), Harderian gland, Meibomian gland, corneal epithelium, bulbar conjunctiva, palpebral conjunctiva, serum, aqueous humor, and lacrimal fluid collected from a secretory duct of ELG. Under the ex vivo assay conditions, ELG and ILG, with or without preincubation, exhibited a cytotoxic effect comparable to that of diluted tears. Lacrimal fluid collected from an ELG duct was similarly effective. These specimens triggered nuclear segmentation that is typical of apoptotic nuclei. All other specimens showed no effect on stromal nuclei under the identical conditions. In some animals, ELG was surgically removed and the tear cytotoxicity was examined in vivo. The tear cytotoxicity in these animals was lost after the surgery, indicating an involvement of ELG, but it was restored 4-24hr afterward, suggesting a compensatory role of ILG.These results suggest that ELG and ILG are the major sources of tear factors that are cytotoxic to the keratocytes in the mouse. PMID- 12907163 TI - Role of arteries in oxygen induced vaso-obliteration. AB - In mice the retinal vasculature develops in the first postnatal week by spreading from the optic nerve head towards the retinal periphery. During this growth period, exposure to hyperoxia causes vaso-obliteration of capillaries in the retinal center but not in peripheral regions. High oxygen levels lead to downregulation of vascular endothelial growth factor (VEGF), an important survival factor for vascular endothelial cells, which could explain the vaso obliteration caused by hyperoxia. However, it is not clear why only capillaries in the center of the retina are affected. We therefore investigated how capillary obliteration correlates with VEGF mRNA distribution by in situ hybridization in retinal whole mount preparations. In mouse pups reared under normoxic conditions VEGF mRNA was detectable across the entire vascular network but was virtually absent in the immediate vicinity of arteries. This was true along developing retinal arteries but also around the optic nerve head through which the entire arterial blood supply for the retinal and hyaloid vasculature passes. In these areas capillaries were absent, resulting in so-called capillary free zones. Exposure to hyperoxia caused an expansion of areas with low VEGF mRNA which correlated with capillary obliteration in these regions. Combined capillary obliteration around the optic nerve head and along retinal arteries lead to a large capillary free zone in the center of the retina. Thus, our observations suggest that hyperoxia affects the retinal vasculature by reducing VEGF mRNA levels near arteries and causing a widening of capillary free zones. PMID- 12907164 TI - Separation of the yellow chromophores in individual brunescent cataracts. AB - Quantitative changes in the 330 nm absorbing chromophores and 350/450 nm fluorophores of water-soluble (WS) and water-insoluble (WI) proteins of individual human cataract lenses were characterized and compared with aged normal human lens. Twenty-five brunescent cataract lenses from India were selected from five different stages (types I-V) based upon the color of the lens. The WS and WI proteins from each lens were collected and subjected to an extensive enzymatic digestion procedure under argon. The lens protein digests were separated by Bio Gel P-2 size-exclusion chromatography and individual peaks were analyzed further by reversed-phase HPLC. The total WI proteins increased and the total WS protein decreased with the development of cataract, especially in the late stages of cataract (III-V). The total 330 nm absorbance and 350/450 nm fluorescence of the WI fraction also increased, however, the A(330) and fluorescence per mg lens protein were constant except for type V (black) lenses. Bio-Gel P-2 chromatography separated the chromophores and fluorophores into four fractions. The main fraction (designated as peak 2+3) from the cataract WI proteins was several times higher than that present in aged normal human lens WI proteins. A significant increase of this fraction was observed in WI proteins, but not in WS proteins with cataract development. Similarly, fractions 1 and 4 in the WI proteins also increased gradually but fraction 5 did not. Reversed-phase HPLC resolved fraction (2+3) of the water-insoluble sonicate supernatant proteins into four 330 nm absorbing peaks and eight fluorescent peaks. Among these peaks, a late-eluting peak (peak 8) increased 10 to 15-fold with the progress of cataract, and accounted for 80% of the total chromophores in type V lenses. This peak may represent limit digests of advanced glycation end-products (AGEs) derived protein cross-links. HPLC profiles of fraction 5 from both WS and WI proteins showed numerous new peaks which were not observed in either WS protein from cataract or WI proteins from aged normal human. The severe coloration and the higher levels of numerous novel chromophores and fluorophores in brunescent cataractous lenses reveal the possibility that a different chemistry occurs during cataract development. PMID- 12907165 TI - Metabolic mapping in mammalian retina: a biochemical and 3H-2-deoxyglucose autoradiographic study. AB - It has long been known that mammalian retinas metabolize glucose aerobically to lactic acid and carbon dioxide. The classical view holds that glucose is the primary substrate for energy metabolism in all retinal cells, and that photoreceptor cells have the highest rates of glycolysis and respiration. A different and more recent view is that the Muller cells are the principal, if not sole aerobic producers of lactate, which then serves as the primary fuel for the mitochondria in photoreceptor cells and other retinal neurons. In this paper, we have examined these two competing hypotheses in rat and guinea pig retinas by identifying the cellular sites of glucose uptake and phosphorylation via hexokinase by means of autoradiographic studies with 3H-2-deoxyglucose (3H-2DG). The rat retina serves as a vascular model and the guinea pig retina serves as an avascular model. Rat and guinea pig eyecups were incubated in oxygenated, bicarbonate-buffered media containing glucose in the presence of labeled and unlabeled 2DG. Biochemical measurements of lactate production and ATP content were made on rat retinas incubated with different concentrations of glucose and 2DG in order to establish the optimal condition for conducting the autoradiographic studies with 3H-2DG. The optimal substrate concentrations were 1mM glucose and 0.25 mM 2DG. Results showed that following incubation of dark adapted rat eyecups for 1 hr in media containing 1mM glucose/0.25 mM 2DG and supplemented with 3H-2DG, the label was distributed throughout all the layers of the retina, from the ganglion cell layer to the retinal pigment epithelium, with denser label associated with the outer retina (photoreceptors) relative to the density of label in the inner retina, as evaluated by counts of silver grains in individual retinal layers. Exposure of rat eyecups to light did not alter the relative distribution of label, but did increase total grain counts by 70%. However, uptake of labeled 2DG, as measured by scintillation counting of radioactivity in trichloroacetic acid extracts, was not significantly different between light- and dark-adapted rat retinas. In guinea pig eyecups, labeled 2DG was distributed throughout all the retinal layers. Addition of 10mM lactate or pyruvate to the glucose/2DG media produced no measurable change in the density or distribution of label in the eyecups. Measurements of the activity of hexokinase in rat retinas revealed that this enzyme was present in both the mitochondrial and cytosolic fractions. The present results suggest that as long as the availability of ambient glucose is adequate, retinal neurons use glucose, rather than glial-derived lactate, as the major substrate for the production of high energy phosphates. PMID- 12907166 TI - Epithelial transdifferentiation and cataract in the human lens. AB - Anterior subcapsular cataracts cause a serious loss of vision and are normally associated with ocular trauma, inflammation or clinical skin conditions. They appear to be accompanied by epithelial cell growth and transdifferentiation where unscheduled production of a number of proteins, including alpha smooth muscle actin (alpha-sma), occurs. Clinical studies have also revealed an up-regulation of the TGFbeta signalling pathway in such cataracts. The present study, using phase contrast and immunofluorescent techniques, was undertaken to investigate the extent of alpha-sma expression in traumatic cataracts, in capsulorhexis specimens obtained during cataract surgery and in aged human lenses from donor eyes. The donor lenses were also exposed to trauma or TGFbeta in culture to observe their relative contribution to alpha-sma production. Dense anterior subcapsular cataracts were relatively rare (<1%), but all showed a pronounced up regulation of alpha-sma, which was located both in anterior cells of normal appearance and in nucleated fibroblastic cells lying beneath the anterior epithelium. Surprisingly, more than 50% of capsulorhexis specimens from mature cataracts showed expression of alpha-sma, although to a limited extent. Alpha-sma was not expressed in any of the clear donor lenses and culture for 8 days in EMEM did not induce expression. Interestingly, unlike their young animal counterparts, human lenses failed to show the presence of alpha-sma when exposed to 10 ng ml( 1) TGFbeta. However, after culture, lenses with pre-existing cortical opacities did express alpha-sma, as did clear lenses subjected to injury or trauma. It appears that the greater the stress, the greater is the expression of alpha-sma. Cataract, and especially cortical cataract, should therefore be seen as associated with stress-induced signalling pathways in the lens that lead to the transdifferentiation of the anterior epithelial cells. PMID- 12907167 TI - Estrogen prevention of lacrimal gland cell death and lymphocytic infiltration. AB - Previous studies have shown that ovariectomy causes necrosis of lacrimal acinar cells, apoptosis of plasma cells and gland lymphocytic infiltration. Both, lacrimal gland cell death and lymphocytic infiltration were prevented by androgen treatment. Since estrogens are removed by ovariectomy, and the synthetic estrogen diethylstilbestrol has been shown to affect some biochemical correlates of lacrimal secretion, the purpose of this study was to determine the effect of 17 beta-estradiol treatment on ovariectomy-induced cell death and lymphocytic infiltration. Sexually mature female New Zealand white rabbits (4-4.5 kg) were ovariectomized and divided into two groups. One group was treated with 0.5 mg kg( 1) per day 17-beta-estradiol, and the other group with vehicle alone. A third group of sham operated rabbits was used as controls and they also were treated with vehicle alone. Six days after surgery, the animals were euthanized, the lacrimal glands removed and processed for analysis of apoptosis as assessed by DNA fragmentation, and for morphological examination. DNA fragmentation was determined using the TUNEL assay and agarose gel electrophoresis. Sections were also stained for rabbit thymic lymphocyte antigen (RTLA), and rabbit CD18. Labelled nuclei and stained areas were quantified by automated densitometry. Ovariectomized rabbits showed a significant increase in the values for degraded DNA as a percent of total nuclear area (2.90+/-0.40%) compared to sham operated rabbits (0.73+/-0.22%). 17-beta-estradiol treatment in ovariectomized rabbits prevented the increase in DNA degradation. Examination of TUNEL assay at higher magnification (40x) confirmed previous studies showing that ovariectomy caused apoptosis of interstitial cells. Significant numbers of bulging cells of very pale appearance under light microscopy, also confirm previously identified necrotic cells in acinar regions. Treatment with 17-beta-estradiol prevented this necrosis. Increased numbers of RTLA(+) and CD18(+) interstitial cells were also evident after ovariectomy. 17-beta-estradiol treatment prevented the increase in the number of lymphoid cells. We confirmed previous observations that suggest that glandular atrophy observed after ovariectomy is likely to proceed by necrosis of acinar cells rather than apoptosis, and that ovariectomy triggers an inflammatory response in the gland. These results suggest that in addition to androgens, estrogens also seem to play a role to maintain lacrimal gland structure and function. A decrease in available estrogen levels could trigger both lacrimal gland apoptosis and necrosis, as well as lymphocytic infiltration. Although, the effect of estrogens in these experiments seems to be direct and not through androgens, the possibility of the role of an autocrine and/or paracrine factors, promoted by estrogen on lacrimal gland cells still needs to be investigated. PMID- 12907168 TI - Characterization of adenovirus p21 gene transfer, biodistribution, and immune response after local ocular delivery in New Zealand white rabbits. AB - Previous studies suggest that local gene therapy with rAd-p21(WAF1/Cip-1) [. Arch. Ophthalmol. 120, (2002) 941-949] may provide an effective adjunctive anti proliferative treatment to prevent glaucoma surgery failure. To further investigate rAd-p21 in this indication, we have characterized several parameters of local gene delivery to conjunctiva including, vector delivery and transgene expression in target tissue, inflammatory response, biodistribution to non-target tissues, and immune response. Quantitative PCR and RT-PCR assays were employed to evaluate rAd-p21 dissemination and gene transfer following a single subconjunctival injection. In target tissue, significant levels of rAd-p21 DNA were found in 6/6 animals 1 and 4 days after injection. rAd-p21 DNA and RNA could be detected in the un-injected contralateral eye but at levels that were 10000 100000 lower than in the injected eye. Expression of human p21 transgene in conjunctival fibroblasts was confirmed by immunohistochemistry. Biodistribution of rAd-p21 following subconjunctival injection was substantially limited to ocular tissue. In 1/6 rabbits, rAd-p21 DNA was found in whole blood, liver, and spleen at levels that were barely detectable. All non-target organs were negative on day 4. In contrast, in a rabbit injected intravenously as a positive control, all blood samples and tissues samples were positive. rAd-p21 delivery to conjunctiva followed by filtration surgery caused an early acute inflammatory response, which by day 14 was indistinguishable from placebo-treated eyes. Neutralizing anti-adenovirus antibodies were detected following administration of rAd-p21 to conjunctiva, however, vector delivery and transgene expression were unaffected in a subsequent administration to the contralateral eye in the same animal. These results show that local delivery to conjunctiva may be a suitable delivery mode for ocular gene therapy. PMID- 12907169 TI - Vasoactive intestinal and calcitonin gene-related peptides, tyrosine hydroxylase and nitrergic markers in the innervation of the rat central retinal artery. AB - The vascular supply of the optic nerve has been studied with different methods including corrosion casts both in humans and in other mammals. In man, primates and some other mammals, such as the rat, a distinct central retinal artery accompanies the optic nerve, and runs through the lamina cribosa to reach the optic nerve head. Similarities between human and rat central retinal artery could serve to understand changes in the autonomic perivascular innervation in glaucoma using the rat as an animal model. Nitric oxide, calcitonin gene-related peptide, neuropeptide Y, substance P and vasoactive intestinal peptide have been identified around the monkey central retina artery. Innervation of the rat central artery, however, has not been described in detail. Using immuno- and histochemical methods, the present study investigates the peptidergic, adrenergic and nitrergic innervation of the rat posterior ciliary artery as well as the central retina artery. Numerous nitric oxide positive nerve fibers were visualized posterior and anterior to the lamina cribosa of the optic nerve. They colocalized with NADPH-diaphorase positive fibers, which could also be observed in two of six specimens studied at the level of the optic nerve head. Calcitonin gene-related peptide, tyrosine hydroxylase, and VIP positive fibers were also observed surrounding the vessels of the rat optic nerve. The presence of neuronal nitric oxide/NADPH-diaphorase and vasoactive intestinal peptide positive nerve fibers surrounding the posterior ciliary and central retinal arteries indicates a vasodilator effect in the rat optic nerve. Tyrosine hydroxylase positive innervation indicates the presence of sympathetic activity, and calcitonin gene related peptide positive fibers indicate sensory innervation by trigeminal primary efferents. PMID- 12907170 TI - AII amacrine cells express the MT1 melatonin receptor in human and macaque retina. AB - AII amacrine cells are critical interneurons in the rod pathway of mammalian retina, active primarily in dim lighting conditions. Melatonin, a neuromodulator produced at night in the retina, is believed to induce retinal adaptation to dim lighting conditions in most vertebrate species examined to date, including humans. We hypothesized that melatonin may influence retinal light adaptation by acting on AII cells directly and thus investigated whether melatonin receptors were expressed in AII neurons. Postmortem nonpathological eyes from four human donors as well as two eyes from two Macaque Fasicularis monkeys were analyzed. Double immunocytochemistry was performed using an anti-MT(1) antibody and an antibody to calretinin, an AII marker. Analysis utilized confocal microscopy. A polyclonal anti-calretinin antibody labelled amacrine cells exhibiting the distinct AII morphology, in both human and macaque retina. MT(1) immunoreactivity in macaque retina was similar to human staining, in that horizontal, amacrine and ganglion cell bodies were stained, as were inner segments of photoreceptors. In human retina 86% of calretinin positive cells expressed the MT(1) receptor peripherally, whereas centrally, 78% colocalization was observed. In the macaque retina, 100% of AII amacrine cells expressed MT(1) immunoreactivity both centrally and peripherally. That virtually all AII neurons express the MT(1) receptor in both human and macaque retina, may provide the first evidence demonstrating a role for melatonin in AII regulation, furthering the hypothesis of melatonin function in retinal light adaptation. PMID- 12907171 TI - Expression of betaA2-crystallin in human lenses. AB - Evidence of betaA2-crystallin expression has been detected in human lenses. The protein, which co-elutes with betaA1/A3 from reversed phase HPLC separation of beta(L)-crystallins, accounts for 1-2% of the lens crystallins. Its molecular mass, M(r) 22 006, is consistent with the cDNA deduced sequence with addition of acetylation at the N-terminal serine residue. Approximately 20% of the protein is phosphorylated at Ser30. PMID- 12907172 TI - Leukocyte rolling and extravasation in surgical inflammation after mechanical and laser-induced trauma in human patients. AB - Acute inflammations main characteristics are exudation of fluid and plasma proteins and emigration of leukocytes, predominantly neutrophils into inflamed tissue. We have previously demonstrated that leukocyte extravasation can be directly monitored in human patients in vivo after tunnel cataract operation, but at the same time we noticed that only certain manipulations trigger also leukocyte extravasation. To define the type of surgical manipulations triggering leukocyte extravasation, we tested a set of surgical manipulations affecting distinct anatomical components of the conjunctiva. This set included various combinations of epithelial, nerve, and/or blood vessel damage. Manipulations tested here were surgical conjunctival biopsy, low-energy Argon laser treatment (80 mJ) closing a venule temporarily, high-energy argon laser treatment (480 mJ) to occlude a venule firmly, and 193 nm eximer laser-based conjunctival phototherapeutic keratectomy (160 mJ). These manipulations were compared to previous findings on conjunctival inflammation following the standard cataract operation. In mechanical trauma models (cataract operation and conjunctival biopsy) clinical signs of inflammation, number of leukocytes rolling, and number of tissue emigrated leukocytes were notably higher compared to pre-operative levels on one day after the operation. No specific anatomical triggering component for inflammation, including epithelial, nerve, and/or blood vessel damage, was indentified, but rather the trauma mechanism itself appeared to be an essential factor. Surprisingly, in laser-induced traumas no increase in number of rolling cells, rolling velocity slowing, or elevation of tissue emigrated leukocytes took place compared to pre-operative levels. These findings suggest that laser-induced traumas differ fundamentally from the mechanical traumas at all levels of leukocyte extravasation cascade, and at least in our setup, inflammatory reaction can be avoided or triggered depending on surgical method used. PMID- 12907173 TI - Face processing in adolescents with autistic disorder: the inversion and composite effects. AB - Two experiments with upright and inverted face and object images were carried out to investigate whether face processing in autism is more feature-based than in individuals with typical development. Participants were 17 high-ability adolescents with autistic disorder (16-24 years), 24 typically developing children (9-10 years) and 16 adults (18-33 years). In Experiment 1, a normal inversion effect was found for the adolescents with autism in a standard face recognition paradigm with reduced memory demands, except for a subgroup with low social intelligence who were not better in recognizing upright relative to inverted photographs of faces. In Experiment 2, the group with autism did not show the composite effect like the adult group did: they recognized face halves as well in aligned composite faces as in non-aligned composite faces. The results on the inversion task suggest that most adolescents with autism form a normal configuration-based face representation, but the absence of the composite effect indicates that they are less prone to use the contextual information of the face in a visual-search task. PMID- 12907174 TI - Effects of interval duration on temporal processing in schizophrenia. AB - INTRODUCTION: Temporal processing has received scant attention in the literature pertaining to cognitive deficits in patients with schizophrenia. Previous research suggests that patients with schizophrenia exhibit temporal perception deficits on both auditory and visual stimuli. The current study investigated the effects of interval manipulation to (1) replicate the original findings with a larger sample and an increased number of trials (2) assess the degree to which both patients and controls can differentiate temporal changes in a range of experimental interstimulus intervals, and (3) explore whether different interstimulus interval durations pose different levels of difficulty for the patients with schizophrenia. METHODS: Participants were asked to decide whether temporal intervals were shorter or longer than standard intervals on a computer based auditory temporal perception task. The standard interval remained the same duration throughout the various tasks. The interstimulus interval separating the standard and experimental intervals varied in the range of 500, 1000, or 3000 ms. Data are presented for a sample of 16 patients with schizophrenia and 15 controls. RESULTS: Data suggest that patients with schizophrenia exhibit deficits in differentiating interval durations across all paradigms compared to their control-group peers on a range of auditory tasks (p<.001). CONCLUSIONS: These results are consistent with a general temporal deficit in schizophrenia. However, the roles of medication and localization are also addressed. PMID- 12907175 TI - Alzheimer's disease disrupts arithmetic fact retrieval processes but not arithmetic strategy selection. AB - Three groups of healthy younger adults, healthy older adults, and probable AD patients, performed an addition/number comparison task. They compared 128 couples of additions and numbers (e.g., 4 + 9 15) and had to identify the largest item for each problem by pressing one of two buttons located under each item. Manipulations of problem characteristics (i.e., problem difficulty and splits between correct sums and proposed numbers) enabled us to examine strategy selection and specific arithmetic fact retrieval processes. Results showed that arithmetic facts retrieval processes, which were spared with aging, were impaired in AD patients. However, AD patients were able to switch between strategies across trials according to problem characteristics as well as healthy older adults, and less systematically than healthy younger adults. We discuss implications of these findings for further understanding AD-related differences in arithmetic in particular, and problem solving in general. PMID- 12907176 TI - Line bisection-based hemisphericity estimates of university students and professionals: evidence of sorting during higher education and career selection. AB - Recently, hemisphericity has been assessed by six intercorrelated methods. Here, one of these, the "Best Hand Test," a two-hand line bisection-based biophysical method relatively independent of language, culture, or education, was applied to the measurement of hemisphericity distributions within large groups. Entering university students (n=402) enrolled in three lower division courses were chosen as a reference population. Each of these classes contained about 56% left brain oriented individuals. In contrast, mean student left-brain distributions in four specialized, upper level courses (n=180), ranged from 35 to 68%, suggestive of hemisphericity sorting. Even more pronounced hemisphericity distribution differences were found within university representatives of 15 professions (n=421) and within professional subspecialists (n=45). For example, of biochemists (n=18), 83% were left brain-oriented, while among astronomers (n=21), only 29% were. These results are of intrinsic interest, and demonstrate the capability of the Best Hand Test to estimate hemisphericity in large groups. PMID- 12907177 TI - The impact of culture and education on non-verbal neuropsychological measurements: a critical review. AB - Clinical neuropsychology has frequently considered visuospatial and non-verbal tests to be culturally and educationally fair or at least fairer than verbal tests. This paper reviews the cross-cultural differences in performance on visuoperceptual and visuoconstructional ability tasks and analyzes the impact of education and culture on non-verbal neuropsychological measurements. This paper compares: (1) non-verbal test performance among groups with different educational levels, and the same cultural background (inter-education intra-culture comparison); (2) the test performance among groups with the same educational level and different cultural backgrounds (intra-education inter-culture comparisons). Several studies have demonstrated a strong association between educational level and performance on common non-verbal neuropsychological tests. When neuropsychological test performance in different cultural groups is compared, significant differences are evident. Performance on non-verbal tests such as copying figures, drawing maps or listening to tones can be significantly influenced by the individual's culture. Arguments against the use of some current neuropsychological non-verbal instruments, procedures, and norms in the assessment of diverse educational and cultural groups are discussed and possible solutions to this problem are presented. PMID- 12907178 TI - Healthy high-hostiles evidence low-alpha power (7.5-9.5Hz) changes during negative affective learning. AB - The present experiment was designed to better understand the impact of positive and negative emotional processing among low- and high-hostile individuals. Based on previous research which found increased sympathovagal balance among low hostiles to the negative version of the Affective Auditory Verbal Learning Test (AAVL), it was hypothesized that low-hostiles would experience increased cortical arousal to this stimulus whereas their high-hostile counterparts would not. As expected, low-hostiles experienced significantly reduced low-alpha power (7.5 9.5Hz) relative to high-hostiles during the presentation of the negative AAVL. In a replication of prior research, significant primacy and recency effects were noted for the negative and positive word lists, respectively. Results are discussed in terms of cerebral activation theory and the potential impact of emotional processing among high-hostile individuals and their likelihood to develop coronary heart disease. PMID- 12907179 TI - The relationship between executive function and verbal memory in Parkinson's disease. AB - A growing body of evidence suggests that the various cognitive symptoms found in Parkinson's disease (PD) are secondary to executive dysfunction. Studies addressing this possibility for memory impairment specifically have not included measures of working memory nor have they ruled-out potential mediating variables such as overall level of cognitive impairment or depression. The purpose of this study was to include measures of these variables in determining the relationship between multiple aspects of executive function and delayed verbal recall in 32 idiopathic PD patients. Results were consistent with the original hypothesis and further suggest that working memory is a key factor in recall memory and may mediate the relationship between other executive measures and recall in PD. PMID- 12907180 TI - Age and responding-hand related changes in performance of neurologically normal subjects on the line-bisection and chimeric-faces tasks. AB - This study investigated age and responding-hand (left, right, and bimanual) related changes in visuospatial attention. Two tasks were completed by 107 neurologically normal right-handed subjects ranging in age from 5 to 70 years and distributed across four age groups. Task-specific differences between groups were apparent. In the line-bisection task, the younger and older groups displayed symmetrical neglect while the young and middle groups displayed pseudoneglect. In the chimeric-faces task the leftward bias was less pronounced in the older group and more susceptible to responding-hand effects in the middle and older groups. Whilst results, especially those of the bimanual method, provided strong support for an activation model, they imposed an age limitation on its appropriateness as an explanation for performance on the chimeric-faces task. Results are discussed as reflecting changes in the corpus callosum and right hemisphere. PMID- 12907181 TI - The role of the executive system in visuo-spatial memory functioning. AB - Participants were presented with a spatial sequence in which between 4 and 10 cells were highlighted. On each trial list length was unknown to the participant who was required to serially recall the last four cells. Processing of longer lists is assumed to call upon the executive system, which is thought to be involved in updating the contents of the short-term visuo-spatial store. Study 1 revealed that loading the executive system with concurrent random letter generation impaired performance on the spatial task especially recall of the early serial positions. However, contrary to expectation the degree of impairment was no greater on the longer lists, where it had been assumed that updating would be occurring. Study 2 confirmed this finding and demonstrated that relative to sequences of known length, under conditions of uncertainty when the list length was unknown, spatial recall was impaired even on short sequences. The present results support the growing consensus, which is suggestive of closer links between visuo-spatial and executive processes. However, it remains unclear whether or not updating is actually occurring on the longer sequences, and if it is, what specific executive processes are involved. PMID- 12907182 TI - Impaired processing of prosodic and musical patterns after right hemisphere damage. AB - The distinction between the processing of musical information and segmental speech information (i.e., consonants and vowels) has been much explored. In contrast, the relationship between the processing of music and prosodic speech information (e.g., intonation) has been largely ignored. We report an assessment of prosodic perception for an amateur musician, KB, who became amusic following a right-hemisphere stroke. Relative to matched controls, KB's segmental speech perception was preserved. However, KB was unable to discriminate pitch or rhythm patterns in linguistic or musical stimuli. He was also impaired on prosodic perception tasks (e.g., discriminating statements from questions). Results are discussed in terms of common neural mechanisms that may underlie the processing of some aspects of both music and speech prosody. PMID- 12907183 TI - Hand laterality and cognitive ability: a multiple regression approach. AB - Several different associations between hand laterality and cognitive ability have been proposed. Studies reporting different conclusions vary in their procedures for defining laterality, and several of them rely on measures which are statistically problematic. Previous methods for measuring relative hand skill have not satisfactorily separated the overall level of hand skill, which is a known correlate of cognitive ability, from the asymmetry of its distribution. This paper uses a multiple regression paradigm that separates these two components. Support is found for Leask and Crow's [Trends in Cognitive Sciences, 5 (2001) 513] proposal that average cognitive ability increases monotonically with increasing strength of laterality, regardless of its direction. The small average advantage to dextrals stems from them being more strongly lateralised than sinistrals. The paucity of strong dextrals amongst the very gifted is due to a smaller variance in cognitive ability in this group. PMID- 12907184 TI - Missing and needed: polar marine ecotoxicology. PMID- 12907185 TI - The precautionary principle and environmental monitoring. PMID- 12907186 TI - Economic and environmental impacts on ports and harbors from the convention to ban harmful marine anti-fouling systems. AB - The recent Diplomatic Conference held (1-5 October 2001) by the International Maritime Organization (IMO) in London adopted the Draft Convention prepared by The Marine Environmental Protection Committee (MEPC) of IMO for the "Control of Harmful Anti-fouling Systems for Ships." The convention has been developed to immediately ban the use of Tributyltin (TBT) globally in anti-fouling paints to "protect the marine environment". The ban on TBT has come about because TBT has detrimental effects on non-target marine organisms. In November 1999, IMO agreed that a treaty be developed by the MEPC to ensure a ban on the application of TBT based anti-fouling paints by 1 January 2003, and a ban on the use of TBT by 1 January 2008. At the meeting surious concern was expressed by some experts for the need to identify in the treaty the necessary regulatory language for: (1) the "safe" removal, treatment, and disposal of marine anti-foulants deemed "harmful" by the treaty and (2) who is liable for the future dredging and disposal of TBT contaminated port and harbor sediments--to also "protect the marine environment". The requirement for "safe" removal and disposal was incorporated at MEPC 46 as Article 5 in the treaty, without it shipyards complying with existing national and local discharge regulations (most have none for discharge of TBT) could inadvertently release more TBT to ports and harbors in the five-year compliance period than has been leached from ships (hulls) in the past 40 years to the same waters. Virginia is the only State in the US that regulates the discharge to below 50 ng/l (50 parts per trillion). However, the liability for the future dredging and disposal costs of TBT-contaminated port and harbor sediments has not been addressed. PMID- 12907187 TI - Distribution and source recognition of polycyclic aromatic hydrocarbons in the sediments of Hsin-ta Harbour and adjacent coastal areas, Taiwan. AB - Thirty-three sediment samples from Hsin-ta Harbour and neighboring coastal areas were analyzed by GC-MS for polycyclic aromatic hydrocarbons (PAHs). Total concentrations of 30 analyzed parental and alkylated PAHs ( summation operator PAH) varied from 98.1 to 3382 ng/g dry weight. MP/P (methylphenanthrenes/phenanthrene) values larger than 2 coincided with very low P/A (phenanthrene/anthracene) values at inner harbour stations, revealing that a significant portion of low molecular weight PAHs are probably from petrogenic pollution sources, specifically, illegal disposal of used motor oil. The 4,6 dimethyldibenzothiophene/3,6-dimethylphenanthrene (4,6-C(2)D/3,6-C(2)P) ratio is found to be more useful than the MP/P ratio in tracing petrogenic PAHs from the inner harbour area to the adjacent coastal environment. In addition, according to hierarchical cluster analysis, collected sediments cluster in three major groups, Off-shore Group, Near-shore Group and Inner Harbour Group. Three diagnostic ratios, 4,6-C(2)D/3,6-C(2)P, PER/ summation operator PAH (perylene to summation operator PAH) and BaA/CHR (benzo(a)anthracene/chrysene), representing petrogenic, biogenic and pyrogenic origins, are found to be effective in differentiating and characterizing sediments among the groups in this study. Enrichment of pyrogenic and petrogenic PAHs in sediments collected exhibits mixing or dilution, spatially, by biogenic (or natural) PAHs. PMID- 12907188 TI - Use of physiological responses in Mytilus trossulus as integrative bioindicators of sewage pollution. AB - Mussels, Mytilus trossulus (average shell length 43+/-0.8 mm), were sampled from a beach in Alaska that received untreated sewage for several years, a second beach adjacent to a secondary wastewater outfall, and two nearby reference beaches. Survival time in air, byssal thread production rate, and prevalence of trematode parasites were determined for each group. Tolerances to aerial exposure was significantly lower (P<0.05) at both sewage outfall sites than at the reference sites. Mussels exposed to untreated sewage produced fewer byssal threads and had a significantly higher prevalence of encysted trematodes than mussels from the other beaches, including the secondary wastewater site. Survival in air, byssal thread production, and trematode prevalence in mussels may be useful indicators in evaluating the longterm health of beaches exposed to sewage. PMID- 12907189 TI - Pollution by polychlorinated biphenyls in an estuary of the Gulf of California. Their toxicity and bioaccumulation in shrimp Litopenaeus vannamei. PMID- 12907190 TI - Can plankton communities be considered as bio-indicators of water quality in the Lagoon of Venice? AB - This study examines whether plankton of the Lagoon of Venice could be considered as a bio-indicator of areas subjected to various anthropogenic influences. This study was a two year hydrochemical and biological survey in five areas of the Lagoon of Venice, each with different environmental conditions due to pollution from urban, industrial, thermal and agricultural wastes. Phytoplankton associations did not show any promising species. In the different lagoonal areas, this community was differentiated into its major groups. In contrast, the copepod Acartia tonsa Dana could be considered as a target species in highly eutrophic areas. PMID- 12907191 TI - The use of a non-lethal tool for evaluating toxicological hazard of organochlorine contaminants in Mediterranean cetaceans: new data 10 years after the first paper published in MPB. AB - In the Mediterranean Sea, top predators, and particularly cetacean odontocetes, accumulate high concentrations of organochlorine contaminants and toxic metals, incurring high toxicological risk. In this paper we investigate the use of the skin biopsies as a non-lethal tool for evaluating toxicological hazard of organochlorines in Mediterranean cetaceans, presenting new data 10 years after the paper published by Fossi and co-workers [Mar. Poll. Bull. 24 (9) (1992) 459] in which this new methodology was first presented. Some organochlorine compounds, now with worldwide distribution, are known as endocrine disrupting chemicals (EDCs). Here the unexplored hypothesis that Mediterranean cetaceans are potentially at risk due to organochlorines with endocrine disrupting capacity is investigated. High concentrations of DDT metabolites and PCB congeners (known as EDCs) were found in the different Mediterranean species (Stenella coeruleoalba, Delphinus delphis, Tursiops truncatus and Balaenoptera physalus). In this paper we also propose benzo(a)pyrene monooxygenase (BPMO) activity in marine mammal skin biopsies (non-lethal biomarker) as a potential indicator of exposure to organochlorines, with special reference to the compounds with endocrine disrupting capacity. A statistically significant correlation was found between BPMO activity and organochlorine levels (DDTs, pp(')DDT, op(')DDT, PCBs and PCB99) in skin biopsies of males of B. physalus. Moreover a statistical correlation was also found between BPMO activity and DDT levels in skin biopsies of the endangered Mediterranean population of D. delphis. These results suggest that BPMO induction may be an early sign of exposure to organochlorine EDCs and can be used for periodic monitoring of Mediterranean marine mammal toxicological status. PMID- 12907192 TI - Natural cleanup of heavy fuel oil on rocks: an in situ experiment. AB - Changes in the chemical composition of a heavy fuel oil, Bunker C, exposed to the elements for 556 days in the vicinity of Brest Harbour (France, (48 degrees 18(') N, 4 degrees 32(') W)) have been studied. Samples with exposure to full or reflected sunlight, and in the dark, were analysed by thin layer chromatography and gas chromatography coupled with mass spectrometry and compared with the initial oil. Using hopane as a conserved internal standard, an average of more than 56% of the total hydrocarbon in the residual stranded oil had been removed in the 556 days. The results indicate that dissolution, biodegradation and photooxidation all play important roles in the weathering process, with their respective contributions depending on the exposure. PMID- 12907193 TI - Field measurements and modeling of dilution in the wake of a US navy frigate. AB - A field measurement and computer modeling effort was made to assess the dilution field of pulped waste materials discharged into the wake of a US Navy frigate. Pulped paper and fluorescein dye were discharged from the frigate's pulper at known rates. The subsequent particle and dye concentration field was then measured throughout the wake by a following vessel using multiple independent measures. Minimum dilution of the pulped paper reached 3.2 x 10(5) within 1900 m behind the frigate, or about 8 min after discharge. Independent measures typically agreed within 25% of one another and within 20% of model predictions. Minimum dilution of dye reached 2.3 x 10(5) at a down-wake distance of approximately 3500 m, or roughly 15 min. Comparison to model measurements were again within 20%. The field test was not only successful at characterizing wake dilution under one set of at-sea conditions, but was successful at validating the computer model used for assessing a wide range of ships and conditions. PMID- 12907194 TI - Turbidity regimes over fringing coral reefs near a mining site at Lihir Island, Papua New Guinea. AB - An extensive sediment transport survey took place at Lihir Island (Papua New Guinea), where mining operations involve disposal of waste rocks and soil in nearshore waters. To investigate the potential impact of these practices over neighbouring fringing reefs, turbidity and sediment accumulation were measured continuously for extended periods. Turbidity records provided a map of observed impact zones based on turbidity thresholds. The main zoning features were (a) that an extreme turbidity gradient persists between the inner harbour (turbidity levels of 100-1000 mg l(-1)) and the adjacent reefs (turbidity levels in the order of 10 mg l(-1)), and (b) that observed zones conform with pre-operations impact predictions. Accumulation measurements unveiled no significant sediment accumulation over fringing coral reefs. This study contributes to the understanding of the potential impact of sediment discharge to nearshore waters. PMID- 12907195 TI - Persistence of petroleum hydrocarbon contamination in sediments of the canals in Venice, Italy: 1995 and 1998. AB - Total hydrocarbon and selected polycyclic aromatic hydrocarbon (PAH) levels were examined in sediments collected from known problematic areas in the canals of Venice, Italy, in 1995 and 1998. Hydrocarbon concentrations were greatest in the interior canals, moderate in the partially enclosed locations and lowest in the open-water sites. Total hydrocarbon and PAH concentrations declined from 1995 to 1998. Ancillary data suggest that this decline may have been in response to the elimination of many industrial activities in the lagoon and to initiating an aggressive canal dredging program. The distributions of individual components were generally similar both years regardless of the total PAH concentration or the location of sample collected. PAH alkyl homolog distributions suggest that atmospheric deposition of petrochemical combustion products is the main source of PAHs to Venice's sediments. In some cases however, the presence of low levels of the two-ring naphthalene homologous series indicate additional low-level inputs of fresh oil. PMID- 12907196 TI - Recovery in rubble fields: long-term impacts of blast fishing. AB - This paper presents initial results from a study of factors that inhibit or enhance hard coral recovery in rubble fields created by blast fishing in Komodo National Park and Bunaken National Park, Indonesia. Within nine sites monitored since 1998, there was no significant natural recovery. Levels of potential source coral larvae were assessed with settlement tiles in the rubble fields and in nearby high coral cover sites. Rubble movement was measured and shown to be detrimental to small scleractinians, especially in high current areas. In shallow water (2-6 m deep), rubble is often overgrown by soft corals and corallimorpharians, which inhibit hard coral survival. There is increased scleractinian recruitment in quadrats cleared of soft coral, and Acropora nubbins transplanted into soft coral fields suffer greater mortality than those transplanted above the soft coral canopy. Gaining an understanding of the prognosis for coral recovery is essential not only in order to assess the long term impacts of blast fishing, but also to improve management decisions about protection of intact reefs and potential restoration of damaged areas. PMID- 12907197 TI - Future aquatic nutrient limitations. AB - Nutrient limitation of phytoplankton growth in aquatic systems is moving towards a higher incidence of P and Si limitation as a result of increased nitrogen loading, a N:P fertilizer use of 26:1 (molar basis), population growth, and relatively stable silicate loading. This result will likely alter phytoplankton community composition, and may compromise diatom-->zooplankton-->fish food webs. PMID- 12907198 TI - Polychlorinated biphenyl, heavy metal and methylmercury residues in hammerhead sharks: contaminant status and assessment. PMID- 12907199 TI - Accumulation of metals in the soft tissues, byssus and shell of the mytilid mussel Perna viridis (Bivalvia: Mytilidae) from polluted and uncontaminated locations in Hong Kong coastal waters. PMID- 12907200 TI - Background concentrations of Cd, Cu, Pb and Zn in the green-lipped mussel Perna viridis (Linnaeus) from Peninsular Malaysia. PMID- 12907201 TI - Management of cancer of the retromolar trigone. AB - Treatment of cancer of the retromolar trigone (RMT) is controversial. While early lesions may be managed with single-modality, more advanced lesions may invade the mandible, pterygoid musculature, and the adjacent mucosa of the tonsillar pillar and soft palate, therefore making therapeutic decisions more complicated. Treatment options traditionally include surgical resection, external beam irradiation, and combined modality therapy. The choice of therapy is dependent on the extent of the tumor, nodal metastasis, and the medical status of the patient and comorbid conditions. The following review outlines the current issues relevant to the diagnosis and therapy of patients with RMT malignancy. PMID- 12907203 TI - Acquisition of anoikis resistance is a critical step in the progression of oral tongue cancer. AB - We hypothesized that acquisition of resistance to anoikis is a critical step in oral cancer progression. To test this hypothesis, we compared a panel of cell lines derived from human oral tissues across the spectrum of tumor progression from oral keratinocytes (HOK-16B), invasive oral squamous cell carcinoma (Tu167), and finally metastatic carcinoma (TxCS-1, MDA1986) for their sensitivity to detachment from the extracellular matrix. The relationship between stage of tumor progression and anoikis resistance was demonstrated by the apoptotic fractions after 48 h in suspension culture which were 93.33, 61.6, 34.5, and 3.71%, respectively. To further demonstrate that anoikis resistance is important for tumor progression, we selected a highly anoikis resistant cell line, JMAR, by serial passage of the Tu167 cell line in suspension culture. Initially, the JMAR line, and clones derived from it, were characterized for anoikis resistance in vitro, and after 72 h in suspension culture the rates of anoikis in the Tu167 and JMAR lines were found to be 73 and 26%, respectively. The degree of anoikis resistance was found to correlate with survival of nude mice orthotopically injected with 5x10(5) Tu167 or JMAR cells. The JMAR mice had a median survival of 17 days versus over 30 days in mice implanted with the Tu167 line. Finally, we found that in vivo selection in the orthotopic model for a regionally metastatic cell line by implantation of Tu167 into the tongues of nude mice and harvesting and culturing cervical lymph nodes led to production of a cell line, Tu167LN1, which was found to be anoikis-resistant. This cell line had an apoptotic cell fraction of 16.2% (+/-3.14%) after 48 h in suspension culture. PMID- 12907202 TI - Quantitative analysis of cathepsin L mRNA and protein expression during oral cancer progression. AB - Although an important risk factor for oral cancer is the presence of epithelial dysplasia, most of these lesions will not progress to malignancy. Presently, for the individual patient with dysplasia, there are few reliable markers that may indicate the likelihood of progression to oral cancer. Cathepsin L is a lysosomal protease that degrades extracellular matrix material. Because cathepsin L is frequently overexpressed in oral squamous cell carcinoma (SCC) we hypothesized that it is also overexpressed in oral premalignancy and that premalignant lesions that progressed to oral cancer expressed higher levels of cathepsin L than those premalignant lesions that did not. In this retrospective pilot study we examined changes in cathepsin L expression at the mRNA level using quantitative TaqMan RT PCR and at the protein level by immunohistochemistry in 33 routinely processed oral dysplastic lesions and 14 SCCs obtained from 33 patients. Sixteen of the dysplastic lesions progressed to oral SCC and 17 did not after several years of follow-up. Cathepsin L mRNA was overexpressed in 16/33 (48%) dysplastic lesions and in 9/14 (64%) oral SCC. Cathepsin L protein was also overexpressed in a large proportion of dysplasias and cancers. Overexpression was independent of dysplasia grade and identified in both those patients who progressed to oral SCC and in those who did not. Levels of cathepsin L mRNA and protein did not differ significantly in the progressing versus non-progressing dysplasias (P=0.27). However, cathepsin L mRNA and protein were significantly lower in the non progressing dysplasias when compared to the oral cancers (P=0.03) but not in the progressing dysplasias suggesting a trend for dysplasias with overexpressed cathepsin L to be more likely to progress to oral cancer. PMID- 12907204 TI - Survival from glottic and supraglottic laryngeal carcinoma in Mumbai (Bombay), India. AB - The survival experience of patients with cancer of the larynx (ICD 10) registered by the Bombay population-based cancer registry India, during the years 1992-1994, is described. The vital status of these subjects were established by matching with death certificates from the Bombay Municipal death register and by active methods such as reply-paid postal enquiries, telephone enquiries, scrutiny of case records and house visits. Of the 675 eligible cases for analysis, 458 (67.9%) were dead and 217 (32.1%) were alive at last follow-up. The 5-year observed and relative survival rates were 29.1 and 38.2%, respectively; these were 52.1 and 58.5% for glottic laryngeal cancer and 24.2 and 31.4% for supraglottic laryngeal cancer, respectively. The 5-year observed survival was 53.1% for those with localised cancer and 17.8% for those with regional extension. Advancing age, regional and metastatic disease and supraglottic cancers were associated with significantly reduced survival. Early detection and prompt treatment should improve overall survival from laryngeal cancer. PMID- 12907205 TI - Socioeconomic status, lifestyle factors and oral premalignant lesions. AB - Several studies have suggested that low socioeconomic status (SES) is associated with a higher risk of oral cancer, but the association with oral premalignant lesions has not yet been explored. The aim of this study was to examine the association of education, occupation, income and SES index with oral premalignant lesions. A case-control study was conducted with data from the baseline screening of a randomized oral cancer screening trial in Kerala, India. There were a total of 927 oral leukoplakia, 170 oral submucous fibrosis, 100 erythroplakia and 115 multiple oral premalignant lesion cases and 47,773 controls. Subjects with high SES index had protective ORs for oral premalignant lesions, ranging from 0.6 to 0.7, after adjustment for age, sex, BMI, tobacco chewing, smoking, drinking and fruit/vegetable intake. Higher education levels were also associated with decreased risk of all four oral premalignant lesions. Protective ORs for income were observed for oral leukoplakia and possibly oral submucous fibrosis and erythroplakia. SES may be associated with oral premalignant lesions because of access to medical care, health related behaviors, living environment or psychosocial factors. Though the mechanism for the association is not clear, higher SES index, education and income were associated with decreased risk of oral premalignant lesions in our study. PMID- 12907206 TI - Immunosuppressive effects of soluble factors secreted by head and neck squamous cell carcinoma on dendritic cells and T lymphocytes. AB - Recent observations suggest that the inability of the immune system to mount an effective immune response against head and neck squamous cell carcinoma (HNSCC) could be a result of the immunosuppression mediated through soluble factors that are secreted by tumour cells. Therefore, we investigated the effects of conditioned medium obtained from cultures of HNSCC cell lines (HNSCC-CM) on the function of dendritic cells (DC) and T cell immune response. In our study, we could not observe any inhibitory effect of HNSCC-CM on the maturation and the cytokine secretion pattern of DC. On the contrary, HNSCC-CM from two of three cell lines consistently decreased the quantity of IFN-gamma- and IL-4-secreting T cells upon restimulation in vitro. In conclusion, our data suggest that soluble factors secreted by HNSCC cells directly inhibit the function of effector T cells, rather than impeding the process of antigen presentation. PMID- 12907207 TI - Treatment of squamous cell carcinoma of the anterior faucial pillar-retromolar trigone. AB - Cancer of the anterior faucial pillar-retromolar trigone is an uncommon head and neck tumor, which has historically been shown to be associated with poor prognosis. In this retrospective study, we reviewed our experience with primary surgery followed by postoperative radiation therapy in order to determine the impact of our treatment protocols on patients' outcome. Between January 1994 and December 1998, 31 patients with histologically proven squamous cell carcinoma (SCC) of the anterior faucial pillar-retromolar trigone were treated in our department. Surgical excision of the primary lesion and ipsilateral neck dissection were performed in all patients. Reconstruction was accomplished using masseter muscle flap or tongue flap. Postoperatively, most patients (90%) received radiation therapy (51-58 Gy) to the primary side and neck. Adjuvant chemotherapy was offered if histologic signs of aggressive behavior were identified. Four out of 31 patients were initially seen at stage I or II and 27 patients at stage III or IV of the disease. Metastatic disease was demonstrated in 78% of ipsilateral neck nodes. Occult metastases were found in 64% of clinically N0 necks. The 3-year loco-regional recurrence rates were 44.8%. SCC of retromolar trigone is considered as an aggressive and insidious tumor. The reconstruction of the deficit of the anterior faucial pillar-retromolar area with masseter muscle flap is a reliable, safe and absolutely functional method. PMID- 12907208 TI - Maxillofacial manifestations of Langerhans cell histiocytosis: a clinical and therapeutic analysis of 10 patients. AB - The definition of Langerhans cell histiocytosis (formerly known as histiocytosis X) includes the clinical syndromes Hand-Schueller-Christian syndrome, Abt Letterer-Siwe syndrome, and eosinophilic granuloma. The paper gives an overview of current diagnostic and treatment strategies of LCH. Furthermore, records and clinical data of 10 patients with LCH were evaluated retrospectively. Patients' age ranged from 13 years to 42 years. The mandible was more frequently involved than the maxilla. Three patients (30%) had systemic manifestations of LCH in addition to their oral lesions. The longest follow-up period was 12 years. During follow-up six patients (60%) developed recurrent LCH and received adjuvant chemo- or radiation therapy. For solitary bone lesions, surgical curettage is the recommended treatment. Those patients with multi-organ involvement or recurrent LCH should be included into clinical trials initiated by the Histiocyte Society. PMID- 12907209 TI - Coffee and tea intake and risk of oral, pharyngeal and esophageal cancer. AB - The relation between coffee, decaffeinated coffee, tea and oral/pharyngeal, and esophageal cancer risk is inadequately quantified. Data were derived from hospital-based case-control studies conducted in Italy and Switzerland. The study on oral/pharyngeal cancer included 749 cases and 1772 controls, and that of esophageal cancer 395 cases and 1066 controls. Multivariate odds ratios (OR) and 95% confidence intervals (CI) were computed. The OR for >3 cups/day of coffee compared with 134.34 and 691.87>166.26 were selected for the quantitation of quinupristin and dalfopristin, respectively. The assay run cycle-time was approximately 2.0 min injection-to-injection. The assay was linear up to concentration of 4000 ng x ml(-1) quinupristin and 1920 ng x ml(-1) dalfopristin. The lowest limits of quantitation of quinupristin and dalfopristin were found to be 1000 and 480 ng x ml(-1), respectively. Quantitation was based on peak area measurement of quinupristin and dalfopristin using weighed linear regression. Linear relationships with correlation coefficients (r>0.99) were automatically computed for both constituents by MASSLYNX quantify program. The ratio of the slopes of the calibration curves of quinupristin and dalfopristin was found to be 0.425, which matches the nominal ratio composition of the antimicrobial compounds in Synercid. The %RSD ranges were 2.3-4.0% for dalfopristin and 1.3-4.2% for quinupristin, whereas the %DEV ranges were (-7.5+3.7) and (-1.2+9.1%), respectively, indicating appropriate precision and accuracy. Recoveries of 99.5 103.8% and 97.8-99.0% of quinupristin and dalfopristin, respectively, were computed from Synercid injection. The described method is recommended for rapid determination of the contents and for tracking the stability and compatibility of quinupristin and dalfopristin in Synercid injection. PMID- 12907261 TI - High-performance liquid chromatographic analysis of bioactive triterpenes in Perilla frutescens. AB - Perilla frutescens (L.) Britt. (Lamiaceae), a famous traditional Chinese medicine, has been used for the treatment of various diseases. To evaluate the quality of P. frutescens, a simple, rapid and accurate high-performance liquid chromatography (HPLC) method was developed for the assessment of three bioactive triterpene acids: tormentic acid (TA), oleanolic acid (OA) and ursolic acid (UA). The HPLC system used an Spherisob octadecylsilyl silica (ODS) column with acetonitrile and aqueous H(3)PO(4) as the mobile phase and detection at 206 nm. The method was precise with relative standard deviations for these three constituents that ranged between 0.6-1.5% (intraday) and 0.7-2.6% (interday). The content of these three phytochemicals in the leaves of P. frutescens growing at eight different locations of China was determined to establish the effectiveness of the method. PMID- 12907262 TI - Evaluation of stability of cefuroxime axetil in solid state. AB - The effect of temperature and relative atmospheric humidity on the stability of the crystalline form of cefuroxime axetil (CFA) in solid state was investigated. CFA is a mixture of diastereomers A and B. Changes in the concentration of the two diastereomers (A and B) of CFA were recorded by means of HPLC with UV detection. The degradation of diastereomers of CFA occurring at 0% relative humidity (RH) of the ambient air is a reversible first order reaction, while that occurring in humid air (RH>50%) is an autocatalytic first order reaction relative to substrate concentration. Although it has been found, that diastereomer B is the more stable isomer, humidity has a stronger effect on this very diastereomer. PMID- 12907263 TI - A rapid and sensitive LC/MS/MS assay for quantitative determination of digoxin in rat plasma. AB - Digoxin is a cardiac glycoside that is widely used for the treatment of congestive heart failure. To evaluate pharmacokinetics of digoxin in rats, a sensitive LC/MS/MS assay was developed and validated for the determination of digoxin concentration in rat plasma. For detection, a Sciex API3000 LC/MS/MS with atmospheric pressure ionization (API) mass spectrometry turbo ion spray inlet in the positive ion-multiple reaction monitoring mode was used to monitor precursor- >product ions of m/z 798.6-->651.6 for digoxin and m/z 577.6-->433.3 for oleandrin, the internal standard (IS). The standard curve was linear (r(2)>or=0.999) over the digoxin concentration range of 0.1-100 ng/ml in plasma for digoxin. The mean predicted concentrations of the quality control samples deviated by <5.8% from the corresponding nominal values; the intra-assay and inter-assay precision of the assay were within 8.6% relative standard deviation. At the lower limit of quantitation (LLQ) of 0.1 ng/ml, the mean deviation of predicted concentrations from the nominal value was within 3.7%. The extraction recoveries of digoxin and internal standard were 82.7+/-3.9 and 105.9+/-2.3%, respectively. The present method was successfully applied to characterization of pharmacokinetic profiles of digoxin in rats after oral administration. PMID- 12907264 TI - Analysis for residual host cell proteins and DNA in process streams of a recombinant protein product expressed in Escherichia coli cells. AB - Analyses of crude samples from biotechnology processes are often required in order to demonstrate that residual host cell impurities are reduced or eliminated during purification. In later stages of development, as the processes are further developed and finalized, there is a tremendous volume of testing required to confirm the absence of residual host cell proteins (HCP) and DNA. Analytical tests for these components are very challenging since (1). they may be present at levels that span a million-fold range, requiring substantial dilutions; (2). are not a single component, often existing as fragments and a variety of structures; (3). require high sensitivity for final steps in process; and (4). are present in very complex matrices including other impurities, the product, buffers, salts and solvents. Due to the complex matrices and the variety of potential analytes, the methods of analysis are not truly quantitative for all species. Although these limitations are well known, the assays are still very much in demand since they are required for approval of new products. Methods for final products, described elsewhere, focus on approaches to achieve regulatory requirements. The study described herein will describe the technical rationale for measuring the clearance of HCP and DNA in the entire bioprocessing to purification from an Escherichia coli-derived expression system. Three analytical assays, namely, reversed-phase high-performance liquid chromatography (RP-HPLC), enzyme-linked immunosorbent assay (ELISA), and Threshold Total DNA Assay, were utilized to quantify the protein product, HCP and DNA, respectively. Product quantification is often required for yield estimation and is useful since DNA and HCP results are best expressed as a ratio to product for calculation of relative purification factors. The recombinant E. coli were grown to express the protein of interest as insoluble inclusion bodies (IB) within the cells. The IB were isolated by repeated homogenization and centrifugation and the inclusion body slurry (IBS) was solubilized with urea. After refolding the product, the solution was loaded on several commonly used ion exchangers (CM, SP, DEAE, and Q). Product was eluted in a salt gradient mode and fractions were collected and analyzed for product, HCP and DNA. The IBS used for this study contained about 15 mg/ml product, 38 mg/ml HCP and 1.1 mg/ml DNA. Thus, the relative amounts of HCP and DNA in the IBS was excessive, and about 10(3) times greater than typical (because the cells and IB were not processed with the normal number of washing steps during isolation). This was of interest since similar samples may be encountered when working with non-inclusion body systems, such as periplasmic expressions, or in cases where the upstream unit operations under-perform in IB cleaning. The study described herein describes the development of three robust methods that provide the essential process data needed. These findings are of general interest to other projects since applications of similar analytical technology may be used as a tool to develop processes, evaluate clearance of impurities, and produce a suitable product. PMID- 12907265 TI - Determination of nifedipine in human plasma by solid-phase extraction and high performance liquid chromatography: validation and application to pharmacokinetic studies. AB - Nifedipine, a dihydropyridine calcium channel antagonist, is widely used in the treatment of hypertension and other cardiovascular disorders. A simple, rapid, sensitive, precise and accurate HPLC method, using solid-phase extraction, for the quantitation of nifedipine in human plasma was developed and validated. The calibration graphs were linear in the 5-400 ng/ml concentration range (r>0.999). Recovery for nifedipine was greater than 93.9% and for internal standard nitrendipine was 96.1%. Intra-day and inter-day precision ranged from 1.4 to 4.2 and 3.9 to 5.6%, respectively. Intra-day and inter-day accuracy was ranged from 94.5 to 98.0 and 93.1 to 98.0%, respectively. The method was not interfered with by other plasma components and was applied for the determination of nifedipine in pharmacokinetic study after single oral administration of 10 mg nifedipine to 18 healthy male subjects. PMID- 12907266 TI - Fluorimetric determination of some antibiotics in raw material and dosage forms through ternary complex formation with terbium (Tb(3+)). AB - A highly sensitive and specific fluorimetric method was developed for the determination of cefazolin sodium I, cefoperazone sodium II, ceftriaxone sodium III, and cefixime IV. The proposed method involves the formation of ternary complex with Tb(3+) in the presence of Tris buffer. The quenching of the terbium fluorescence due to the complex formation was quantitative for the four studied drugs. The effect of pH, concentration of Tris buffer and terbium were studied. The formation of the complex was highly dependent on the pH. The optimum pH was found to be pH 8 for cefazolin sodium I, ceftriaxone sodium III, cefixime IV and pH 10 for cefoperazone sodium II. The optimum concentration for Tb(3+) was found 1 ml of 10(-4) M solution and for Tris buffer 1 ml of the prepared solution. Under the described conditions, the proposed method was applicable over the concentration range 8.79 x 10(-6)-7.91 x 10(-5), 9.7 x 10(-6)-4.49 x 10(-)5, 6.10 x 10(-6)-2.50 x 10(-5), and 4.92 x 10(-6)-2.95 x 10(-5) mol with mean percentage accuracy of 99.79+/-0.24, 98.97+/-1.25, 100.05+/-0.79, and 100.15+/-0.54 for I, II, III, and IV, respectively. The proposed method was applied successfully for the determination of studied drugs in bulk powder and in pharmaceutical formulations. The results obtained by applying the described method were statistically analyzed and compared with those obtained by applying the official method. The proposed method was used as stability indicating method for the determination of the studied drugs in the presence of their degradation products. PMID- 12907267 TI - Voltammetric investigation of diethylstilbestrol. AB - In this work electrooxidation of diethylstilbestrol (DES) was investigated by cyclic voltammetry (CV) and differential pulse voltammetry (DPV) using a glassy carbon (GC) electrode. It was statistically shown that both methods could be used for the determination of DES in the concentration range of 2 x 10(-5)-6 x 10(-4) M by CV and 1 x 10(-5)-1 x 10(-3) M in methanol (MeOH) and 4 x 10(-5)-6 x 10(-4) M in acetonitrile (ACN) by DPV and both of the methods could be applied to human serum. A mechanism was proposed about the electrooxidation of this substance. PMID- 12907268 TI - Physicochemical compatibility between ketoprofen lysine salt injections (Artrosilene) and pharmaceutical products frequently used for combined therapy by intravenous administration. AB - Ketoprofen lysine salt (Artrosilene) Fiale) is a non-steroidal anti-inflammatory agent frequently administered by intravenous infusion in association regimen with other drugs, such as steroidal anti-inflammatory, anti-hemorrhagic, anti-spastic, anti-ulcer, and antibacterial drugs. The aim of this study was to investigate the physicochemical compatibility between ketoprofen lysine salt (Artrosilene) Fiale) and other injectable drugs frequently used in association. Physicochemical properties of ketoprofen lysine salt mixtures with different drugs, including colour, clarity, pH and drug content were observed or measured before and after (up to 5 h) mixing at room temperature and under light protection. Results show that the association of Artrosilene Fiale with different drugs does not cause, up to 5 h from mixing, any significant variation in the physicochemical parameters mentioned above. In conclusion, the results obtained demonstrated the physicochemical compatibility of ketoprofen lysine salt (Artrosilene) Fiale) with several drugs. PMID- 12907269 TI - Purkinje cell vulnerability and autism: a possible etiological connection. AB - Autism is a neurological disorder of unknown etiology. The onset of the abnormal growth and development within the brain is also not known. Current thought by experts in autism is that the time of onset is prenatal, occurring prior to 30 weeks gestation. However, autism comprises a heterogeneous population in that parents report either that their child was abnormal from birth, or that their child was developmentally normal until sometime after birth, at which time the child began to regress or deteriorate. Anecdotal reports suggest that some children with autism have significant illness or clinical events prior to the development of autistic symptoms. Conceivably, these children may become autistic from neuronal cell death or brain damage sometime after birth as result of insult. To support this theory is that marked Purkinje cell loss, the most consistent finding in the autistic disorder, can result from insult. Evidence suggests that the Purkinje cell is selectively vulnerable. This article discusses a theory that the selective vulnerability of the Purkinje cell may play a role in the etiology of autism, and suggests that a future direction in autism research may be to investigate the possibility of neuronal cell loss from insult as a cause of autism. Results of a small pilot survey are also discussed. PMID- 12907270 TI - ADHD: topic update. PMID- 12907271 TI - Benign childhood epilepsy with centrotemporal spikes: a study of 50 Chinese children. AB - In order to collect local data for children with Benign Childhood Epilepsy with Centrotemporal Spikes (BECTS), we conducted a retrospective study of 50 Chinese children (32 males and 18 females) with BECTS diagnosed in two regional hospitals in Hong Kong from 1995 to 1998. Their peak age of onset was 7 years (range 3-13 years) and a male predominance was observed. Seven patients (14%) had a past history of febrile convulsions and five cases (10%) had a family history of epilepsy. The presentation was protean, but most of them had infrequent, short, nocturnal generalised seizures. The EEG spike foci were most frequently found in mid-temporal regions, followed by centrotemporal regions. Fourteen percent of children did not require anti-epileptic drug treatment. For those who were treated, they were easily controlled on a low dose of carbamazepine (median dosage of 12.75 mg/kg per day) or sodium valproate (median dosage of 20 mg/kg per day). Our study suggested a generally good prognosis for BECTS. No risk factors of frequent seizure recurrence could be identified. PMID- 12907272 TI - Visual and auditory event related potentials in epileptic children: a comparison with normal and abnormal MRI findings. AB - Visual and auditory event related potentials (VERPs and AERPs) in 32 epileptic children with magnetic resonance imaging (MRI) abnormalities and 18 with normal MRI were recorded and compared to the data of 21 healthy children. Of all 50 epileptic children (34 male, 16 female) aged 14.42+/-4.27 (7-20) years, 21 were medically intractable and receiving polytherapy. The mean latencies of N2 and P3 components of VERPs and AERPs in all epileptic children were significantly higher than those of the controls (P<0.05). Epileptic children with structural abnormalities had more prolonged latencies of N2 and P3 components of AERPs and VERPs than those of the healthy children (P<0.05). The epileptic children with normal MRI had significantly more prolonged latency of N2 and P3 of VERPs and P3 of AERPs than those of the controls (P<0.05). The difference of the mean latency of N2 and P3 components or the mean amplitude of P3 components of ERPs between the two epileptic groups was insignificant. The type of medication (mono- versus polytherapy) did not affect the wave components of ERPs. We suggest that epileptic activity, itself, leads to prolonged N2 and P3 components of AERPs and VERPs. The presence of structural abnormality indicated by imaging is not a predictor of ERPs abnormalities. PMID- 12907273 TI - Paroxysmal movement disorders in severe myoclonic epilepsy in infancy. AB - We report on the electroclinical findings and the results of a molecular genetic study of a patient with typical severe myoclonic epilepsy in infancy (TSME) and three with borderline SME (BSME) who showed paroxysmal movement disorders, such as choreoathetosis, dystonia and ballismus, during their clinical course. BSME was defined as a clinical entity that shares common characteristics with TSME but lacks myoclonic seizures associated with ictal EEG changes. When the paroxysmal movement disorders were first observed, all the patients in this study were being treated with polytherapy including phenytoin (PHT), and these abnormal movements disappeared when PHT was discontinued or reduced. However, on other occasions, two of our cases also showed the same abnormal movements even when not being treated with PHT. One patient with TSME and two of the three patients with BSME had SCN1A gene mutations that lead to truncation of the associated protein. We conclude that paroxysmal movement disorders seen in SME patients were closely related to their AED therapy, especially the use of PHT. It is thought that patients with both TSME and BSME have some predisposition toward paroxysmal movement disorders, and that this predisposition is partly related to sodium channel dysfunction, although some other factors might influence the occurrence of this phenomenon. PMID- 12907274 TI - Amplitude spectral analysis of maturational changes of delta waves in preterm infants. AB - The aim of this study is to clarify the usefulness of amplitude spectral analysis for an evaluation of maturational changes of delta activities in preterm infants. We chose each ten healthy infants without complications who were 29-30, 31-32, and 33-34 weeks of post-conceptional age (PCA) at electroencephalogram (EEG) recordings. Fast Fourier transform algorithm was applied for amplitude spectral analysis. The analyzed data were divided into four frequency bands; D1 0.53-1, D2 1-2, D3 2-3, and D4 3-4Hz. The average amplitude of six segments with high voltage slow waves was calculated in each frequency band. A significant reduction of the amplitude along with PCA was present in all leads in D1 band. On the other hand, a significant negative correlation with PCA was observed only in the occipital leads in D2, D3 or D4 bands. In conclusion, maturational EEG changes assessed by amplitude spectral analysis were prominent in the occipital areas, and in the frequency less than 1Hz. PMID- 12907275 TI - In vivo quantitative ultrasonic evaluation of neonatal brain with a real time integrated backscatter imaging system. AB - We applied the integrated backscatter (IBS) imaging system to the evaluation of the normal neonatal brain of different birth-weights: extremely low-birth-weight (N=13), very-low-birth-weight (N=14), low-birth weight (N=14), and normal birth weight (N=19). The IBS values in six regions of interest, the deep white matter, subcortical white matter, choroid plexus, thalamus, lateral ventricle, and occipital bone, were compared among groups of different birth weights, gestational age, and postnatal age: at the date of birth and 28 approximately 30 days after birth. The IBS values were higher in the order of bone>choroid plexus>deep white matter>subcortical white matter>thalamus>lateral ventricle and were significantly different except for the lateral ventricle in all the groups at days 0 and 28 approximately 30. The IBS values increased with the decrease of birth weight and gestational age. There was a decrease of IBS values at day 28 compared to that of day 0 in the extremely low birth weight and very low birth weight groups; however, they remained the same in infants with low birth weights and in the normal birth weight group. Further studies to evaluate the significance of this technique for the objective diagnosis of brain insults in neonates are necessary. PMID- 12907276 TI - Spontaneous intracranial haemorrhage in children: aetiology, presentation and outcome. AB - We are aware of only few reports addressing spontaneous intracranial haemorrhage (SICH) in non-selected series of children. This is a retrospective analysis of clinical presentation and outcome in 34 children with SICH seen in the period 1990-2000 in our hospital. Traumatic, neonatal and subdural haemorrhages were excluded. The majority (47%, 16/34) of SICH was caused by an arteriovenous malformation (AVM). SICH was presented as an acute event in 53% (17/32) of patients. Forty-seven percent (15/32) of the cases had protracted courses, which made the diagnosis difficult. In our series, younger children had less specific symptoms. Mortality (25%, 8/32) and morbidity were considerable, particularly in infratentorial haemorrhages, in aneurysms, in children younger than 3 years and in those with underlying haematological disorders. PMID- 12907277 TI - Absence seizures in patients with localization-related epilepsy. AB - We studied the clinical features of 12 patients with localization-related epilepsy (LRE) associated with absence seizures (AS). AS did not appear in any patients before partial seizures (PS) were first observed. The interval between the onset of PS and AS ranged from 1 month to 7.2 years (mean 2.11 years). The duration of AS (mean 5 months) was short compared with that of PS (mean 3.8 years). Carbamazepine (CBZ) was used in seven patients at the onset of AS. It was discontinued in five but continued in the other two. AS was initially treated with valproate in ten patients. Three of them needed additional antiepileptic drugs: clonazepam in two patients and ethosuximide in one. All patients became free from AS after treatment for AS was started, whereas PS was relatively intractable. Generalized spike-and-waves were often observed before the onset of AS. The interval between the first appearance of generalized spike-and-waves and the onset of AS ranged from 1 to 53 months (mean 20 months). AS in patients with LRE will be relatively benign and transient, and will respond well to antiepileptic drugs. PMID- 12907278 TI - Neonatal seizures: characteristics of EEG ictal activity in preterm and fullterm infants. AB - The neonatal EEG remains one of oldest, yet most valuable, diagnostic and prognostic tests in neonates. The goals of this study were to determine the relationships between the morphology, frequency, and distribution of ictal discharges in the neonatal EEG with age, EEG background activity, and etiology. A total of 156 ictal events were evaluated in 11 preterm (PT) and 25 fullterm (FT) infants. Most of the infants had severe abnormalities of background activity although ictal discharges occurred on both normal and abnormal backgrounds. There was a trend for a closer relationship between behavioral changes during the electroencephalographic seizure when the background activity was normal or moderately abnormal than when background activity was severely abnormal. In both PT and FT infants, the most common site of seizure origin was the temporal lobe. FT infants commonly had sharp waves, spikes, sharp and slow waves, and spike and slow waves at the onset of the ictus while rhythmic delta activity was most common in the PT infants. PT infants typically had a regional onset to the ictus whereas FT infants most frequently had a focal onset. Duration of the ictal events was similar in PT and FT infants and a change in morphology or frequency of the discharges was common during propagation of the ictal discharges in both age groups. There was not a clear relationship between onset, morphology, frequency, or propagation patterns and etiology in either the PT or FT infants. Our results demonstrate that while the type of ictal discharge is related to gestational age, there is a rich variety in the onset, morphology, and frequency of the ictal discharges in both PT and FT infants and that neonatal ictal patterns lack a close correlation with underlying pathology. PMID- 12907279 TI - A 1-year-old infant with McArdle disease associated with hyper-creatine kinase emia during febrile episodes. AB - A 14-month-old girl was hospitalized due to repeated hyper-creatine kinase (CK) emia during pyrexia. Mild hypotonia was observed, but other physical and neurological findings were unremarkable. The serum CK level was normal at rest or normothermia. Open muscle biopsy was performed on the rectus femoris, and showed glycogen storage and complete lack of phosphorylase activity histochemically and biochemically, establishing the diagnosis of McArdle disease. The diagnosis of McArdle disease in early infancy is uncommon. Until this study there have been no reports of clinical symptoms or muscle biopsy findings for McArdle disease in early childhood. This disease must be considered when transient hyper-CKemia is observed in infants, even if glycogen storage is unremarkable as compared with adult cases. PMID- 12907280 TI - Ataxia with vitamin E deficiency and severe dystonia: report of a case. AB - Mutation of the gene for alpha-tocopherol transfer protein causes ataxia with isolated vitamin E deficiency, a disorder usually stabilized or improved after vitamin E supplementation. Dystonia has rarely been described in ataxia with isolated vitamin E deficiency (AVED) patients. We present the case of a young boy with AVED, whose neurological and extra-neurological cardinal symptoms of the disease improved after vitamin E supplementation but who progressively developed generalized dystonia. PMID- 12907281 TI - Focal polymicrogyria, continuous spike-and-wave discharges during slow-wave sleep and Cohen syndrome: a case report. AB - Cohen syndrome is a rare genetic disorder consisting of truncal obesity, hypotonia, mental retardation, characteristic facial appearance and ocular anomalies. Other diagnostic clinical features include narrow hands and feet, low growth parameters, neutropenia and chorioretinal dystrophy. Here, we report an 18 year-old male with Cohen syndrome associated with focal polymicrogyria and continuous spike-and-wave discharges during slow-wave sleep. PMID- 12907282 TI - NCNP News. PMID- 12907284 TI - Effect of intragranular porosity on compression behaviour of and drug release from reservoir pellets. AB - In this study, reservoir pellets were prepared and their compression behaviour as well as the importance of their porosity for compression-induced changes in drug release was investigated. Pellets of three different porosities, consisting of microcrystalline cellulose and salicylic acid, were prepared by extrusion spheronisation and spray-coated with ethyl cellulose (ethanol solution). Lubricated reservoir pellets were compressed and retrieved by deaggregation of the tablets. The retrieved pellets were analysed regarding porosity, thickness, surface area, shape and drug release. It was found that the coating did not significantly affect their compression behaviour. Compaction of pellets of high original porosity considerably affected densification and degree of deformation, whereas the effect on drug release was minor. For low porosity pellets the influence of compaction on drug release was appreciable, but only slight regarding densification and degree of deformation. In conclusion, the porosity of pellets is a potential factor that the formulator can use to optimize drug release and one that can affect the robustness of a formulation during manufacture. Moreover, the coating may be able to adapt to the densification and deformation of the pellets. PMID- 12907285 TI - Evaluation of microcrystalline chitosans for gastro-retentive drug delivery. AB - In vivo absorption studies were carried out in human volunteers to evaluate whether microcrystalline chitosan (MCCh) granules would be gastro-retentive. Furosemide, which is site-specifically absorbed from the upper gastrointestinal tract, was used as model drug. The rate of release of furosemide in vitro could be prolonged by increasing the molecular weight (M(w)) or amount of MCCh (150 to 240 kDa; 80 to 95%) in the granules, and also by addition of acidic excipients to the formulations. No marked changes in the in vivo absorption rate (t(max)) were noted, but the amounts of furosemide absorbed (AUC(0- infinity ) and C(max)) decreased as the in vitro release rate decreased, although this was not statistically significant in the case of AUC. The highest AUC(0- infinity ) (3050 micro g l(-1) h) for furosemide (40 mg) was achieved with granules containing 80% MCCh of 150 kDa M(w). With MCCh of 240 kDa M(w) AUC(0- infinity ) was 1890 micro g l(-1) h. This kind of pharmacokinetic profile of furosemide suggests that the gastric retention time of the granules is too short in relation to the release rate, and a large amount of the drug passes its "absorption window" before being released. The in vivo study produced no evidence that the chitosan formulations studied can be used as mucoadhesive gastro-retentive drug delivery systems. The results of in vitro mucoadhesion studies did not predict the results of in vivo studies. PMID- 12907286 TI - In vivo pharmacokinetics in human volunteers: oral administered guar gum-based colon-targeted 5-fluorouracil tablets. AB - The objective of the present study is to compare the guar gum-based colon targeted tablets of 5-fluorouracil against an immediate release tablet by in vitro dissolution and in vivo pharmacokinetic studies in human volunteers. Twelve healthy volunteers participated in the study. 5-Fluorouracil was administered at a dose of 50 mg both in immediate release tablet and colon-targeted tablet. On oral administration of colon-targeted tablets, 5-fluorouracil started appearing in the plasma at 6 h, and reached the peak concentration (C(max) of 216+/-15 ng/ml) at 7.6+/-0.1 h (T(max)), whereas the immediate release tablets produced peak plasma concentration (C(max) of 278+/-21 ng/ml) at 0.6+/-0.01 h (T(max)). The AUC(0- infinity ) for 5-fluorouracil from colon-targeted tablet and immediate release tablet were found to be 617+/-39 and 205+/-21 ng/ml/h, respectively. Colon-targeted tablets showed delayed t(max), delayed absorption time (t(a)), decreased C(max) and decreased absorption rate constant when compared to the immediate release tablets. The results of the study indicated that the guar gum based colon-targeted formulation did not release the drug in stomach and small intestine, but delivered it to the colon resulting in a slow absorption of the drug and making it available for local action in colon. PMID- 12907287 TI - Enhanced film-forming properties for ethyl cellulose and starch acetate using n alkenyl succinic anhydrides as novel plasticizers. AB - PURPOSE: The aim of this study was to investigate the ability of n-alkenyl succinic anhydrides (n-ASAs) to improve the film-forming characteristics of a novel coating polymer, potato starch acetate degree of substitution 2.8 (SA). n ASAs were also applied to improve the otherwise brittle properties of ethyl cellulose (EC) aqueous dispersion (Aquacoat) and EC solvent-based films. METHODS: The effectiveness of two n-ASAs, 2-octenyl succinic anhydride (OSA) and 2-dodecen 1-ylsuccinic anhydride were evaluated as plasticizers. Mechanical properties, both water vapor and drug permeabilities, and glass transition temperatures of the cast free films were measured. Triethyl citrate and dibutyl sebacate were used as reference plasticizers. RESULTS: The long hydrocarbon chain of n-ASA, with its accessible carbonyl groups, enabled a strong plasticization effect on the tested polymers. Due to the excellent mechanical properties (i.e., a tough film structure with considerable flexibility) and low permeability of the plasticized films, n-ASAs, and especially OSA proved to be an ideal plasticizer particularly for EC based coatings. Also, the EC aqueous dispersion plasticized with n-ASAs resulted in a markedly enhanced coalescence of the colloidal polymer particles, even at low drying temperatures. CONCLUSIONS: In applications where a coating with high flexibility is required, n-ASAs can be used as plasticizers at moderately high concentrations (up to 60-70%, w/w) without losing the high tensile strength, excellent toughness and low permeability of EC and SA films. PMID- 12907288 TI - Relation between structural characteristics of talc and its properties as an antisticking agent in the production of tablets. AB - Antisticking power varies according to the talc considered. Besides its chemical properties, it is necessary to assess its physical properties related to functionality. It is difficult to define the physical properties of talc implicated in its antisticking power. In this work, different talcs were characterised and their performance in reducing sticking in tablet manufacturing was evaluated. The following parameters were studied: apparent density, morphogranulometry, roughness, and the specific surface through the adsorption desorption of argon. Next, the relationship between the characteristics of talcs and their antisticking power was considered. Talc before and after delamination which is a way to obtain talcs with different physical characteristics-was compared. Antisticking power appeared to be dependent on the basal dimensions of talc, and on the ratio value of the external specific surface measured by diffractometry to the total specific surface by the BET method. Models to express the effect of textural factors of talc particles on antisticking power were defined. PMID- 12907289 TI - Investigation of melt agglomeration process with a hydrophobic binder in combination with sucrose stearate. AB - The melt agglomeration process of lactose powder with hydrogenated cottonseed oil (HCO) as the hydrophobic meltable binder was investigated by studying the physicochemical properties of molten HCO modified by sucrose stearates S170, S770 and S1570. The size, size distribution, micromeritic and adhesion properties of agglomerates as well as surface tension, contact angle, viscosity and specific volume of molten HCO, with and without sucrose stearates, were examined. The viscosity, specific volume and surface tension of molten HCO were found to be modified to varying extents by sucrose stearates which are available in different HLB values and melt properties. The growth of melt agglomerates was promoted predominantly by an increase in viscosity, an increase in specific volume or a decrease in surface tension of the molten binding liquid. The agglomerate growth propensity was higher with an increase in inter-particulate binding strength, agglomerate surface wetness and extent of agglomerate consolidation which enhanced the liquid migration from agglomerate core to periphery leading to an increased surface plasticity for coalescence. The inclusion of high concentrations of completely meltable sucrose stearate S170 greatly induced the growth of agglomerates through increased specific volume and viscosity of the molten binding liquid. On the other hand, the inclusion of incompletely meltable sucrose stearates S770 and S1570 promoted the agglomeration mainly via the reduction in surface tension of the molten binding liquid with declining agglomerate growth propensity at high sucrose stearate concentrations. In addition to being an agglomeration modifier, sucrose stearate demonstrated anti adherent property in melt agglomeration process. The properties of molten HCO and melt agglomerates were dependent on the type and concentration of sucrose stearate added. PMID- 12907290 TI - Dissolution testing of acetylsalicylic acid by a channel flow method-correlation to USP basket and intrinsic dissolution methods. AB - A new modification of the channel flow dissolution method is introduced together with the theoretical basis to extract the solubility and mass transfer parameters from the dissolution experiments. Correlation of drug dissolution profiles in the channel flow apparatus was evaluated with respect to USP basket and intrinsic dissolution methods at pH 1.2 or 6.8. Acetylsalicylic acid (ASA) was studied as a pure drug substance and as three simple tablet compositions with microcrystalline cellulose (MCC) and/or lactose as excipients. The channel flow measurements of 100% ASA tablets correlated well with the results of intrinsic dissolution tests. In the channel flow method as well as in the USP basket method the release of ASA was fastest from the tablet compositions containing lactose, while the slowest dissolution rate was observed with the composition containing MCC as the only excipient. As presumed, the dissolution rate of the weak acid was decreased as the pH of the medium was lowered, which was clearly confirmed also by the three dissolution methods. MCC forms matrix tablets and in the USP basket method the dissolution profiles followed square root of time kinetics indicating that diffusion was the rate-controlling step of ASA dissolution. Also the channel flow results indicated that the dissolution of ASA was controlled by mass transfer. The swelling behaviour of the tablets is different in the channel flow method as compared to the basket method: only one tablet surface is exposed to the dissolution medium in the channel flow system. The contact between the tablet surface and the dissolution medium is more similar between the channel flow and intrinsic dissolution methods. PMID- 12907291 TI - Pharmacological evaluation of several major ingredients of Chinese herbal medicines in human hepatoma Hep3B cells. AB - Long-dan-tan (Chinese name) is one of the most common herbal medicines used by Chinese people with chronic liver disease. Accumulated anecdotal evidence suggests that Long-dan-tan may show a beneficial effect in patients with hepatocellular carcinoma. Long-dan-tan is made from five plants: Gentiana root, Scutellaria root, Gardenia fruit, Alisma rhizome, and Bupleurum root. In this study, we have examined the cytotoxic effects of the five major ingredients isolated from the above plants, i.e. gentiopicroside, baicalein, geniposide, alisol B acetate and saikosaponin-d, respectively, on human hepatoma Hep3B cells. Annexin V immunofluorescence detection, DNA fragmentation assays and FACScan analysis of propidium iodide-staining cells showed that gentiopicroside, baicalein, and geniposide had little effect, whereas alisol B acetate and saikosaponin-d profoundly induced apoptosis in Hep3B cells. Alisol B acetate, but not saikosaponin-d, induced G2/M arrest of the cell cycle as well as a significant increase in caspase-3 activity. Interestingly, baicalein by itself induced an increase in H(2)O(2) generation and the subsequent NF-kappaB activation; furthermore, it effectively inhibited the transforming growth factor beta(1) (TGF-beta(1))-induced caspase-3 activation and cell apoptosis. We suggest that alisol B acetate and saikosaponin-d induced cell apoptosis through the caspase-3-dependent and -independent pathways, respectively. Instead of inducing apoptosis, baicalein inhibits TGF-beta(1)-induced apoptosis via increase in cellular H(2)O(2) formation and NF-kappaB activation in human hepatoma Hep3B cells. PMID- 12907292 TI - Characterization of a new rat urinary metabolite of piperine by LC/NMR/MS studies. AB - Potential of piperine, an active alkaloid of black and long peppers, to increase the bioavailability of drugs in humans is of great clinical significance owing to its omnipresence in food. In an attempt to further study the reported differences in its metabolism in rats and humans, a new major urinary metabolite was detected in rat urine and plasma using HPLC. The metabolite was partially purified using reverse phase column chromatography on Sephadex((R))-LH 20 and characterized as 5 (3, 4-methylenedioxy phenyl)-2E,4E-pentadienoic acid-N-(3-yl propionic acid) amide with the help of LC/NMR/positive ESI-MS studies. Complete mass fragmentation pattern could be assigned with MS/MS studies. The metabolite has a unique structure compared to the previously reported metabolites in that it retains methylenedioxy ring and conjugated double bonds while the piperidine ring is modified to form propionic acid group. Mechanism of formation of the metabolite by oxidation and cleavage of piperidine ring is proposed. Kidney appears to be the major excretion route for piperine metabolites in rats as no metabolite could be detected in feces. PMID- 12907293 TI - Thermodynamics of solutions. II. Flurbiprofen and diflunisal as models for studying solvation of drug substances. AB - Three independent methods (sublimation, solubility and solution calorimetry) were used to study the dissolution and solvation processes of diflunisal (DIF) and flurbiprofen (FBP). Thermodynamic functions for the sublimation of DIF and FBP were obtained. Concentrations of saturated solutions and standard solution enthalpies of DIF and FBP in aliphatic alcohols and individual organic solvents were measured. Correlation analysis between: (a) the thermodynamic functions for a substance in various solvents, and (b) the same functions for different compounds was carried out. The investigated substances can be arranged with increasing Gibbs energy of solvation as follows: benzoic acid/ =0.6+/-0.06D/f and polarizability, Tr(Deltaalpha;) approximately 55+/-5 A(3)/f(2) upon electronic excitation, which are similar to those of unbound monomeric Chl a, indicating the absence of strong delocalization of the excitations which would be expected in the presence of strong excitonic interactions. The Stark effect in the Chl b absorption region is significantly bigger with /Deltamu-->/ values of the order of 2.0+/-0.2 D/f and it is attributed to strong interactions with neoxanthin molecules. Clear oligomerisation-dependent differences are observed in the carotenoid region, mainly due to the appearance of a new xanthophyll absorption band at 509 in the spectra of trimers and oligomers. It is ascribed to some lutein molecules, in agreement with previous experimental observations. The electro-optic properties of these lutein molecules are significantly different from those of the other xanthophylls in LHCII, which do not exhibit such a big change in dipole moment upon electronic excitation (/Deltamu-->/ =14.6+/-2.0 D/f). Upon aggregation of LHCII some extra absorption appears on the red side of the main Chl a Q(y) absorption band. In contrast to an earlier suggestion [J. Phys. Chem., A 103 (1999) 2422], no indications are found for the charge-transfer character of the corresponding band. The assignments of the S(2) electronic transitions of neoxanthin and lutein in LHCII and possible origins of the Stark effect are discussed. PMID- 12907304 TI - Chlorophylls of the c family: absolute configuration and inhibition of NADPH:protochlorophyllide oxidoreductase. AB - Using circular dichroism (CD) spectroscopy, the stereochemistry at C-13(2) of members of the chlorophyll (Chl) c family, namely Chls c(1), c(2), c(3) and [8 vinyl]-protochlorophyllide a (Pchlide a) was determined. By comparison with spectra of known enantiomers, all Chl c members turned out to have the (R) configuration, which is in agreement with considerations drawn from chlorophyll biosynthesis. Except for a double bond in the side chain at C-17, the chemical structure of Chl c(1) is identical with Pchlide a, the natural substrate of the light-dependent NADPH:protochlorophyllide oxidoreductase (POR). Thus, lack of binding to the active site due to the wrong configuration at C-13(2), which had been proposed previously, cannot be an explanation for inactivity of Chl c in this enzymic reaction. Our results show rather that Chl c(1) is a competitive inhibitor for this enzyme, tested with Pchlide a and Zn-protopheophorbide a (Zn Ppheide a) as substrates. PMID- 12907305 TI - "Presynaptic silence" may be golden. AB - Conversion of "silent" synapses into active ones is a likely mechanism for long term potentiation (LTP), an experimental paradigm for studying information storage. A widely accepted mechanism that has been suggested for synaptic silence is that functional AMPA glutamate receptors (AMPARs) are absent on the subsynaptic membrane. Evidence is presented here that in many cases the cause of apparent "silence" is presynaptic and due to a low level of glutamate release. Increased transmitter release is crucial for early LTP maintenance. Delayed modifications in postsynaptic receptors matched with transmitter release changes underlie structural alterations associated with late LTP phases. PMID- 12907306 TI - Postsynaptic silent synapses: evidence and mechanisms. AB - In this review I discuss the evidence that some glutamatergic synapses exist that lack surface-expressed postsynaptic AMPA receptors (AMPARs) but contain NMDA receptors opposed to a functional release site. I have summarised the electrophysiological, anatomical and cell biological evidence for such postsynaptically silent synapses, and data that support the idea of rapid AMPAR insertion at silent synapses during long-term potentiation (LTP). I also discuss recent findings suggesting that developmental and activity-dependent alteration in the postsynaptic glutamate receptor composition is a general principle that occurs for other receptor subtypes. This review is not intended to provide a full discussion of possible presynaptic mechanisms for silent synapses; these are covered in the accompanying recent article [Voronin and Cherubini (this issue)]. PMID- 12907307 TI - Steroid-sensitive GABAA receptors in the fetal sheep brain. AB - Neuroactive steroids such as allopregnanolone (3 alpha-hydroxy-5 alpha-pregnan-20 one) influence central nervous system (CNS) excitability by increasing GABA (gamma aminobutyric acid) inhibitory activity. Allopregnanolone concentrations are higher in the fetal compared to the adult ovine brain, suggesting that this neurosteroid may have a role in regulating fetal CNS activity during gestation. We examined the localisation of allopregnanolone-sensitive GABA(A) receptors in the fetal brain to determine if their sensitivity to allopregnanolone changed during late gestation. The binding of [(35)S] tert-butylbicyclophosothionate (TBPS) was used to identify the GABA-chloride ion receptor complex in fetal sheep brains at 90-95, 115-120 and 140-145 days gestational age (GA; term approximately 147 days), by autoradiography. Allopregnanolone (200 nM) reduced [(35)S]TBPS binding by 70-100% throughout the brain at all fetal ages examined. The levels of [(35)S]TBPS binding increased with advancing gestation in all regions examined except some areas of the medulla. Functionally related nuclei and brain areas associated with regulating somato/viscerosensory functions displayed high levels of [(35)S]TBPS binding from mid-gestation. These results indicate that allopregnanolone may interact with GABA(A) receptors to inhibit fetal CNS activity from mid-gestation. This inhibition may contribute to maintaining the sleep-like behaviour and low incidence of arousal-type activity typical of fetal life, and may be neuroprotective by limiting excitatory neurotransmission. PMID- 12907308 TI - Systemic administration of the potent mGlu8 receptor agonist (S)-3,4-DCPG induces c-Fos in stress-related brain regions in wild-type, but not mGlu8 receptor knockout mice. AB - The effect of a novel and potent metabotropic glutamate 8 (mGlu8) receptor agonist, (S)-3,4-dicarboxyphenylglycine (DCPG), was studied in vivo in mouse brain. c-Fos expression was used as a marker of neuronal activity in specific brain regions 2 h after systemic (S)-3,4-DCPG (3-100 mg/kg, i.p.). The selectivity of (S)-3,4-DCPG on mGlu8 receptors was determined in mGlu8 receptor knockout mice. In wild-type mice, (S)-3,4-DCPG (100 mg/kg) significantly increased c-Fos expression in several stress-related brain regions: paraventricular nucleus of the hypothalamus, central nucleus of the amygdala, lateral parabrachial nucleus and locus coeruleus. In the central nucleus of the amgydala, more than 92% of c-Fos positive neurons were identified as GABAergic inhibitory neurons after (S)-3,4-DCPG. Moreover, (S)-3,4-DCPG significantly induced c-Fos in the superficial gray layer of the superior colliculus, a central visual region. c-Fos expression was unchanged by (S)-3,4-DCPG in mGlu8 receptor knockout mice. Our results indicate that systemic (S)-3,4-DCPG alters neuronal excitability in specific brain regions via mGlu8 receptor. The prominent activation of stress areas suggests a role for mGlu8 receptors in the central integration of stress responses. Furthermore, our results indicate that systemic (S)-3,4-DCPG can be used as a tool to explore behavioral and cellular consequences of mGlu8 receptor activation. PMID- 12907309 TI - Neuroprotective action of MPEP, a selective mGluR5 antagonist, in methamphetamine induced dopaminergic neurotoxicity is associated with a decrease in dopamine outflow and inhibition of hyperthermia in rats. AB - The aim of this study was to examine the role of metabotropic glutamate receptor 5 (mGluR5) in the toxic action of methamphetamine on dopaminergic neurones in rats. Methamphetamine (10 mg/kg sc), administered five times, reduced the levels of dopamine and its metabolites in striatal tissue when measured 72 h after the last injection. A selective antagonist of mGluR5, 2-methyl-6 (phenylethynyl)pyridine (MPEP; 5 mg/kg ip), when administered five times immediately before each methamphetamine injection reversed the above-mentioned methamphetamine effects. A single MPEP (5 mg/kg ip) injection reduced the basal extracellular dopamine level in the striatum, as well as dopamine release stimulated either by methamphetamine (10 mg/kg sc) or by intrastriatally administered veratridine (100 microM). Moreover, it transiently diminished the methamphetamine (10 mg/kg sc)-induced hyperthermia and reduced basal body temperature. MPEP administered into the striatum at high concentrations (500 microM) increased extracellular dopamine levels, while lower concentrations (50 100 microM) were devoid of any effect. The results of this study suggest that the blockade of mGluR5 by MPEP may protect dopaminergic neurones against methamphetamine-induced toxicity. Neuroprotection rendered by MPEP may be associated with the reduction of the methamphetamine-induced dopamine efflux in the striatum due to the blockade of extrastriatal mGluR5, and with a decrease in hyperthermia. PMID- 12907310 TI - Caffeine and muscarinic antagonists act in synergy to inhibit haloperidol-induced catalepsy. AB - The possible synergism between caffeine and muscarinic antagonists to inhibit haloperidol-induced catalepsy was investigated with the bar test in rats. Pretreatment with low doses of caffeine (1-3 mg/kg), a non-selective adenosine antagonist, dose dependently reduced the intensity and increased the onset latency of catalepsy induced by haloperidol (0.5-2 mg/kg). Similar effects were produced by the muscarinic antagonists atropine (4.1 mg/kg), and trihexyphenidyl (THP, 0.01-3 mg/kg). THP inhibited catalepsy intensity with an ED(50) of 0.38 mg/kg, and increased its onset latency with an ED(50) of 0.52 mg/kg. The anticataleptic effect of anticholinergics was potentiated when a low dose of caffeine (1 mg/kg) was applied simultaneously. In the presence of caffeine, THP inhibited catalepsy intensity with an ED(50) of 0.19 mg/kg, and prolonged the latency with an ED(50) of 0.30 mg/kg. The synergism was more evident when THP was administered at subthreshold doses that were unable to modify haloperidol-induced catalepsy when applied alone, but produced a clear inhibition of catalepsy when injected with caffeine. To assess whether repeated administration of caffeine could induce tolerance to the synergism with THP, a group of rats was pretreated with three daily doses of caffeine (1 mg/kg) for seven days, and the catalepsy test was performed on the eighth day. In these animals, caffeine was still able to enhance the anticataleptic actions of THP, suggesting that repeated administration of 1 mg/kg caffeine does not induce tolerance to the synergism with anticholinergics. These results indicate that low doses of caffeine enhance the anticataleptic actions of muscarinic antagonists, and leave open the possibility of using caffeine as adjunctive therapy to reduce the doses and the adverse effects of anticholinergics in Parkinson's disease. PMID- 12907311 TI - Muscarinic acetylcholine receptors potentiate the GABAergic transmission in the developing rat inferior colliculus. AB - Postsynaptic currents (PSCs) were recorded using the patch-clamp technique in neurons of the rat inferior colliculus (IC) to investigate the muscarinic modulation of the GABAergic transmission. In the presence of strychnine (0.5 microM) and kynurenic acid (1 mM), spontaneous GABAergic PSCs were observed in all IC neurons investigated. Muscarine (10 microM) greatly potentiated the frequency of these GABAergic PSCs (618% of the control). 4-DAMP (50 nM), a M3 receptor preferring antagonist, greatly blocked the muscarine-evoked PSC frequency increase. The muscarinic antagonists telenzepine (50 nM; M1 preferring), methoctramine (10 microM; M2 preferring), and himbazine (10 microM; M4 preferring), and the nicotinic antagonist mecamylamine (10 microM) did not significantly affect the muscarine effect. These findings indicate that the muscarinic modulation of the GABAergic transmission is primarily mediated by M3 receptors, while M1-, M2- and M4- and nicotinic receptors do not participate substantially. In the presence of tetrodotoxin (0.5 microM), muscarine failed to increase the PSC frequency indicating that its effect depended on the generation of spikes. We suggest that GABAergic interneurones express M3 receptors at some distance from the terminal. Their activation excites GABAergic interneurones, thereby enhancing GABA release in the IC. The muscarinic modulation of the GABAergic transmission may play an important role in the maturation of inhibitory synapses in the developing IC. PMID- 12907312 TI - Norepinephrine release in amygdala of rats during chronic nicotine self administration: an in vivo microdialysis study. AB - The essential role of the amygdala in learning and memory, including cue associated learning, is influenced by local release of norepinephrine (NE). The current study investigated changes in amygdaloid NE secretion in rats learning to self-administer nicotine in an unlimited access model (23 h/day). In vivo microdialysis of NE was performed for 9 h intervals during three phases of nicotine self-administration: acquisition (day 1); early maintenance, when self administration rates first stabilized (day 8.4+/-0.7); and later, during fully stable maintenance (day 17.6+/-1.0). On day 1, a greater number of self administration episodes (SAEs) were associated with elevated NE levels in rats bar-pressing for nicotine (88% vs. 39% with saline). By early maintenance, such episodes increased threefold and overall NE levels were greater. During later maintenance, however, bar-pressing behavior was similar and NE was elevated by the first SAE of the day, but total daily NE levels were no longer elevated. In all the three phases, the enhanced NE release during the first daily SAE did not occur in the last SAE 9 h later. Thus, in an animal model of unlimited nicotine self-administration that approximates the human pattern of nicotine consumption via smoking, the amygdaloid NE response to nicotine diminishes over each day and with the stabilization of self-administration. The decline of amygdaloid NE secretion after long-term nicotine self-administration likely reflects desensitization to the pharmacological effects of nicotine. In addition, amygdaloid NE release, which enhances the consolidation of amygdala-dependent memory, may no longer be necessary once self-administration behavior has been established. PMID- 12907313 TI - L-DOPA increases noradrenaline turnover in central and peripheral nervous systems. AB - The ability of L-3,4-dihydroxyphenylalanine (L-DOPA) to release noradrenaline (NA) from peripheral and CNS neurons was studied using isolated rat vas deferens and in vivo frontal cortex microdialysis. Application of L-DOPA (30 microM) to vas deferens increased basal NA efflux but not electrical field stimulation evoked release of NA when the tissue was pretreated with an inhibitor of MAO-B (clorgyline 1 microM) or an inhibitor of MAO-A and MOA-B (AGN-1133 1 microM). No effect on NA efflux was seen when the tissue was treated with rasagiline (0.005 microM; selective inhibitor of MAO-B), but rasagiline, AGN-1133 and clorgyline increased basal efflux of dopamine (DA) following L-DOPA. In microdialysis experiments, systemic administration of L-DOPA/carbidopa combination (50/12 mg.kg(-1)) increased the efflux of 3,4-dihydroxyphenylglycol and 3-methoxy,4 hydroxyphenylglycol but reduced that of NA. Microdialysate levels of NA, however were increased following L-DOPA/carbidopa when desipramine (1 microM) was infused locally via the probe, or following systemic administration of yohimbine (2 mg.kg(-1)). The data are consistent with the hypothesis that administration of L DOPA is followed by increased axoplasmatic levels of DA which release NA from storage sites into the free cytoplasmatic pool. PMID- 12907314 TI - Agmatine exerts anticonvulsant effect in mice: modulation by alpha 2 adrenoceptors and nitric oxide. AB - The effect of agmatine, an endogenous polyamine metabolite, on seizure susceptibility was investigated in mice. Acute intraperitoneal administration of agmatine (5, 10, 20, 40 mg/kg) had a significant and dose-dependent inhibitory effect on pentylenetetrazole (PTZ)-induced seizures. The peak of this anticonvulsant effect was 45 min after agmatine administration. We further investigated the possible involvement of the alpha(2)-adrenoceptors and L arginine/NO pathway in this effect of agmatine. The alpha(2)-adrenoceptor antagonist, yohimbine (0.5-2 mg/kg), induced a dose-dependent blockade of the anticonvulsant effect of agmatine. The nitric oxide synthase (NOS) substrate, L arginine (60 mg/kg), inhibited the anticonvulsant property of agmatine and this effect was significantly reversed by NOS inhibitor N(G)-nitro-L-arginine (L-NAME, 30 mg/kg), implying an NO-dependent mechanism for L-arginine effect. We further examined a possible additive effect between agmatine (1 or 5 mg/kg) and L-NAME (10 mg/kg). The combination of L-NAME (10 mg/kg) with agmatine (5 but not 1 mg/kg) induced a significantly higher level of seizure protection as compared with each drug alone. Moreover, a combination of lower doses of yohimbine (0.5 mg/kg) and L-arginine (30 mg/kg) also significantly decreased the anticonvulsant effect of agmatine. In conclusion, the present data suggest that agmatine may be of potential use in seizure treatment. PMID- 12907315 TI - Intrathecal nerve growth factor restores opioid effectiveness in an animal model of neuropathic pain. AB - It is without dispute that the treatment of neuropathic pain is an area of largely unmet medical need. Available analgesics, such as morphine, either have minimal effects in neuropathic pain patients, or are not always well tolerated due to concurrent adverse effects. The chronicity of neuropathic pain is thought to be related to many neurochemical changes in the dorsal root ganglia (DRG) and spinal cord, including a reduction in the retrograde transport of nerve growth factor (NGF). In this study, we have determined the ability of chronic intrathecal (i.t.) infusion of NGF to reverse neuropathic pain symptoms and to restore morphine's effectiveness in an animal model of neuropathic pain. Seven days after sciatic nerve constriction injury, NGF was administered to the spinal cord by continuous infusion (125 ng/microl/h) via osmotic pumps attached to chronically implanted i.t. catheters. Spinal infusion of NGF did not affect the expression of tactile allodynia or thermal (hot) hyperalgesia in neuropathic rats, although it significantly increased cold water responses frequency at day 14. Following infusion of vehicle, i.t. morphine (20 microg) was ineffective in altering somatosensory thresholds in neuropathic rats. In contrast, morphine substantially attenuated the neuropathy-induced warm and cold hyperalgesia, as well as tactile allodynia, in neuropathic rats chronically infused with i.t. NGF. In addition, we demonstrate that i.t. morphine-induced antinociception was augmented by a cholecystokinin (CCK) antagonist in animals chronically infused with i.t. antibodies directed against NGF. We hypothesize that NGF is critical in maintaining neurochemical homeostasis in the spinal cord of nociceptive neurons, and that supplementation may be beneficial in restoring and/or maintaining opioid analgesia in chronic pain conditions resulting from traumatic nerve injury. PMID- 12907316 TI - Differential regulation of brain derived neurotrophic factor transcripts by antidepressant treatments in the adult rat brain. AB - Antidepressants are known to increase brain derived neurotrophic factor (BDNF) mRNA in the adult rat brain. The BDNF gene has four differentially regulated promoters that generate four transcript forms, each containing a unique non coding 5' exon (exon I-IV) and a common 3' coding exon. Using in situ hybridization with exon-specific riboprobes, we have examined whether diverse classes of antidepressants recruit a single or multiple BDNF promoters to regulate BDNF mRNAs. The antidepressants tested were electroconvulsive seizure (ECS) and the pharmacological antidepressants tranylcypromine, desipramine and fluoxetine. The effects of both acute and chronic ECS were the most prominent on exon I and II containing BDNF mRNAs in hippocampal and cortical subfields. Chronic ECS enhanced the acute induction of exon I, II and IV mRNAs but did not influence the acute upregulation of exon III mRNAs. Acute pharmacological antidepressants resulted in region-specific decreases in distinct exon-specific BDNF transcripts. In contrast, chronic administration with tranylcypromine and desipramine enhanced exon II and exon III mRNAs, respectively, in discrete hippocampal and cortical subfields. Chronic fluoxetine treatment did not have a significant effect on the exon-specific BDNF transcripts. The results indicate that distinct antidepressants differentially regulate BDNF mRNAs through a region specific recruitment of the four BDNF promoters and suggest that diverse signaling mechanisms may be recruited to regulate BDNF transcripts. PMID- 12907317 TI - An invited commentary. Wall thickening in the GI tract on CT, what does it mean? PMID- 12907318 TI - The evolution of ascitic fluid analysis in the diagnosis of spontaneous bacterial peritonitis. PMID- 12907319 TI - Aminosalicylates and colorectal cancer in IBD: a not-so bitter pill to swallow. AB - Inflammatory bowel disease (IBD) is associated with an increased risk of developing intestinal cancer at sites of chronic inflammation. Aminosalicylates, including both sulfasalazine and mesalamine, are the most commonly prescribed anti-inflammatory agents prescribed in IBD. On balance, the body of literature to date suggests that aminosalicylates confer some protection against the development of colonic neoplasia in patients with IBD and in a variety of models, including in the noninflamed gut. This latter observation implies that aminosalicylates may be of chemopreventive value in normal as well as IBD individuals. The current review examines and gives an overview of the evidence from a variety of sources, including epidemiological, in vivo and in vitro studies that have investigated the potential anticancer effects of aminosalicylates. PMID- 12907320 TI - Effects of gender and age on esophageal biomechanical properties and sensation. AB - OBJECTIVES: The effects of gender and age on human esophageal function is poorly defined. Our aim was to investigate the sensory perception and the biomechanical properties of the esophagus both at the smooth muscle and the striated muscle segments in age- and gender-matched healthy subjects. Eleven older individuals (five men and six women, aged 55-82 yr) and 11 younger healthy individuals (five male and six female, aged 22-45 yr) participated in this study. METHODS: Graded balloon distentions were performed at the striated muscle and the smooth muscle segments of the esophagus in a random order using impedance planimetry. Impedance planimetry facilitated simultaneous assessments of the biomechanical properties and sensory perception. RESULTS: We found that both at the striated and smooth muscle segments, the cross-sectional area, circumferential wall tension-strain relationship and the sensory thresholds were similar between men and women. In contrast, at both of these segments, the cross-sectional area was larger (p < 0.05), the circumferential wall tension-strain association had shifted significantly (p < 0.05) to the left (i.e., the wall was stiffer), and the median thresholds for discomfort and pain were higher (p < 0.05) in older subjects. The biomechanical properties and thresholds for sensory perception also varied between the two esophageal segments. CONCLUSIONS: Aging was associated with a larger lumen and a stiffer but less sensitive esophageal wall. Thus, aging but not gender may influence esophageal function. PMID- 12907321 TI - Effect of prolonged gastric distention on lower esophageal sphincter function and gastroesophageal reflux. AB - OBJECTIVES: Morbidly obese patients treated with an intragastric balloon report a transient increase in gastroesophageal reflux (GER) symptoms. In the present study, we evaluated the underlying mechanisms of GER and examined the effect of prolonged gastric distention on lower esophageal sphincter function. METHODS: Fasting and postprandial manometric studies were performed in obese subjects (n = 15) before, immediately after, and 10 and 20 wk after placement of a 500-ml water filled balloon. RESULTS: Residual lower esophageal sphincter (LES) pressure after water swallows was not affected after balloon placement, excluding mechanical interaction with sleeve function. Postprandial LES pressure was significantly increased after 10 and 20 wk. GER was increased in the right recumbent position until 10 wk after balloon placement, mainly because of an increased percentage of transient lower esophageal sphincter relaxations (TLESRs) accompanied by GER. TLESRs were the main mechanisms underlying reflux both before and after balloon placement. The rate of TLESRs was increased significantly immediately after introduction of the balloon, returning to baseline values after 20 wk. After balloon placement, reflux episodes were evoked by gastric contractions that were not inhibited by meals. CONCLUSIONS: Chronic distention by an intragastric balloon increased reflux up to 10 wk after placement because of an increase in the percentage of TLESRs accompanied by a reflux episode. In addition, prolonged balloon distention increased the rate of TLESRs and created a postprandial state even 10 wk after balloon placement. After 20 wk these effects largely resolved, illustrating adaptation to this artificial situation. PMID- 12907322 TI - Hypertrophy of the muscularis propria of the lower esophageal sphincter and the body of the esophagus in patients with primary motility disorders of the esophagus. AB - OBJECTIVES: Patients with diffuse esophageal spasm (DES) and nutcracker esophagus/high amplitude esophageal contraction (HAEC) have a thicker esophageal muscularis propria than do healthy subjects. The goals of this study were to determine the esophageal muscle cross-sectional area (MCSA), a measure of muscle mass, in patients with achalasia of the esophagus; and to compare it with that in patients with DES, patients with HAEC, and normal subjects. METHODS: Using a high frequency ultrasound probe catheter, concurrent manometry and ultrasound images of the esophagus were recorded in four subject groups: normal volunteers, patients with HAEC, patients with DES, and patients with achalasia of the esophagus. Recordings were obtained from the lower esophageal sphincter (LES) and multiple sites in the esophagus 2, 4, 6, 8, and 10 cm above the LES. RESULTS: The LES and esophageal muscle thickness as well as esophageal MCSA were greater in all three patient groups than in the normal subject group. Muscle thickness and MCSA were observed to be greatest in patients with achalasia, which were greater than in patients with DES, which were greater than in those with HAEC, which in turn were greater than in normal subjects. CONCLUSIONS: We propose that an increase in the MCSA is an important feature of patients with primary motility disorders of the esophagus. The degree of increase in muscle mass may be an important determinant of the type and the severity of esophageal motor dysfunction. PMID- 12907323 TI - Influence of age and gender on gastric acid secretion as estimated by integrated acidity in patients referred for 24-hour ambulatory pH monitoring. AB - OBJECTIVES: Earlier studies have suggested that gastric acid secretion declines with age. Integrated acidity (IA) calculated from ambulatory 24-h pH studies provides an estimation of gastric acid secretion that is less cumbersome than nasogastric aspiration. The aim of this study was to calculate IA as a function of age and gender. METHODS: In a retrospective study, 753 ambulatory 24-h pH studies from 1994 to 2001 were analyzed. Subjects were classified according to nine age groups: <35 yr (n = 124), 35-39 yr (n = 90), 40-44 yr (n = 92), 45-49 yr (n = 111), 50-54 yr (n = 96), 55-59 yr (n = 61), 60-64 yr (n = 59), 65-69 yr (n = 42), and > or =70 yr (n = 64). Basal integrated acidity (BIA) was defined as IA between 12 midnight and 8 AM when no meals were consumed. Daytime and nocturnal IA/h was measured from 8 AM to 8 PM and from 8 PM to 8 AM. Postprandial IA (PIA) was measured for 2 h after the largest meal peak in each patient. RESULTS: Gender distribution was similar between groups (chi(2) = 6.42, 3 df, p = 0.093). Mean gastric pH was similar between genders (2.96 +/- 1.36 women; 3.01 +/- 1.42 men; p = 0.71). Women and men had similar BIA (361.1 +/- 384.1 vs 434.6 +/- 436.6 mmol/L/h, p = 0.10), daytime IA (33.5 +/- 33.0 vs 34.1 +/- 33.7 mmol/L, p = 0.90) and nocturnal IA (43.3 +/- 42.1 vs 48.0 +/- 46.6 mmol/L, p = 0.42), and PIA (38.2 +/- 58.7 vs 36.0 +/- 58.5 mmol/L/h, p = 0.47). Mean gastric pH, daytime and nocturnal IA, BIA, and PIA did not differ between age groups (p > 0.1). None of the parameters studied had a linear association with age (mean gastric pH, r = 0.106; BIA, r = -0.124; daytime IA, r = -0.046; nocturnal IA, r = -0.104; PIA, r = -0.117). CONCLUSIONS: Based on this study, we conclude that IA is unaffected by gender or age. PMID- 12907324 TI - Colonization with cagA-positive Helicobacter pylori strains in intestinal metaplasia of the esophagus and the esophagogastric junction. AB - OBJECTIVES: Recent studies indicate that colonization with cagA-positive Helicobacter pylori (H. pylori) strains may protect against gastroesophageal reflux disease (GERD) and its complications, but the role of cagA in the etiology of Barrett's esophagus has so far been poorly investigated. The pathogenesis of intestinal metaplasia (IM) at an endoscopically normal esophagogastric junction (EGJ) is still unclear, and the role of the H. pylori virulence factor cagA in it has not been investigated. The aim of our study was to assess the relationship between H. pylori and cagA-positive H. pylori in particular and IM at an endoscopically normal EGJ and Barrett's esophagus. METHODS: Serum samples were obtained from 62 patients without IM, 43 patients with IM at an endoscopically normal junction, and 51 patients with Barrett's esophagus. IM was defined as presence of goblet cells with positive staining with Alcian blue. The prevalence of H. pylori and cagA was investigated by assessment of IgG antibody levels as determined by ELISA. RESULTS: The overall H. pylori prevalence was 59% (92/156), and the cagA prevalence was 29% (46/156). Although 63% (39/62) of IM negative subjects and 74% (32/43) of those with IM at the junction were H. pylori positive, only 41% (21/51) of Barrett's patients tested positive. The differences between the IM negative and the Barrett's group (p = 0.02) and between IM at the junction and Barrett's were significant (p = 0.002). The relative cagA prevalence (percentage with cagA positivity and H. pylori positivity) was 56% (22/39) in patients who were IM negative, 59% (19/32) in those with IM at the junction, and 24% (5/21) in those with Barrett's. The prevalence of anti-CagA was significantly lower in patients with Barrett's esophagus compared with patients who were IM negative (p = 0.002) and those who had IM at the junction (p < 0.001). No difference in cagA prevalence was seen between the latter groups. CONCLUSIONS: These findings are in line with the concept that H. pylori and cagA-positive strains in particular protect against the development of Barrett's esophagus. In contrast, our findings do not support the theory that IM at an endoscopically normal esophagogastric junction is associated with H. pylori or cagA-positive strains. IM at the junction and Barrett's esophagus seem to have different etiologies. PMID- 12907326 TI - Incidental findings of thickening luminal gastrointestinal organs on computed tomography: an absolute indication for endoscopy. AB - OBJECTIVES: Incidental findings of thickened luminal GI organs (LGIO) on CT are not uncommon. However, the significance of these findings is unclear. Because of the lack of scientific data, there are no clinical guidelines for the evaluation of these radiologic abnormalities. Our objective was to determine whether endoscopic evaluation of these findings revealed significant abnormalities. METHODS: This study evaluated all incidental findings of thickened LGIO in a large medical center from October, 1997 to March, 1999 that were followed by endoscopic examinations. RESULTS: Ninety-six percent of patients with incidental findings of thickening of the sigmoid colon or rectum, 81% of patients with thickening of the distal esophagus, and 13% of patients with thickening of the cecum had significant abnormalities on further endoscopic work up. CONCLUSIONS: Although positive pathologic findings are less common in thickening of the cecum than in other LGIO, all of these incidental findings on CT warrant further endoscopic examination. PMID- 12907325 TI - The gastrointestinal safety of the COX-2 selective inhibitor etoricoxib assessed by both endoscopy and analysis of upper gastrointestinal events. AB - OBJECTIVES: Etoricoxib is a selective cyclooxygenase inhibitor that in clinical studies has improved the signs and symptoms of osteoarthritis and rheumatoid arthritis and reduced the potential for GI injury. The incidence of endoscopically detected ulcers and of clinically important upper GI events (perforations, ulcers, and bleeding episodes) was compared in patients taking etoricoxib or nonselective nonsteroidal anti-inflammatory drugs (NSAIDs). METHODS: Upper GI endoscopy was performed at intervals over 12 wk in 680 patients taking etoricoxib 120 mg once daily, ibuprofen 800 mg three times daily, or placebo in a randomized, parallel-group, double-blind study. Survival analysis was used to analyze time-to-event data for the incidence of gastric or duodenal ulcers (> or =3 mm and > or =5 mm), and the log rank test was used to compare the cumulative incidence between treatment groups. A combined analysis of upper GI events in all 10 Phase II/III clinical trials of etoricoxib (60, 90, or 120 mg) versus nonselective NSAIDs (naproxen, ibuprofen, or diclofenac) for osteoarthritis, rheumatoid arthritis, and chronic low back pain was conducted. Investigators reported potential events for adjudication by an external, blinded committee, using prespecified criteria to confirm events. All events that occurred during active treatment periods (maximum 792 days) or within 14 days of stopping treatment were included in the analysis. Time to first event was evaluated using survival analysis; the Kaplan-Meier method was used to determine the cumulative incidence, and relative risk was estimated with the Cox proportional hazards model. RESULTS: In the endoscopy study, the cumulative incidence of ulcers >/=3 mm at 12 wk in the ibuprofen group (17%) was significantly higher than in the etoricoxib group (8.1%, p < 0.001); similar results were seen for ulcers >/=5 mm. In the placebo group, the rate of ulcers >/=3 mm was 1.86%. Of 3142 patients treated with once-daily etoricoxib and 1828 patients treated with a nonselective NSAID (ibuprofen, naproxen, or diclofenac), 82 patients with investigator-reported upper GI events (71 confirmed) were eligible for the combined analysis. For etoricoxib versus NSAIDs, the rate per 100 patient-yr for confirmed events was 1.16 versus 3.05 (relative risk = 0.44, 95% CI = 0.27-0.72, p < 0.001), whereas that for investigator-reported events was 1.35 versus 3.42 (relative risk = 0.47, 95% CI = 0.30-0.74, p = 0.001). Results were driven primarily by studies with naproxen as the comparator. CONCLUSIONS: The incidence of endoscopically detected ulcers was significantly lower with etoricoxib 120 mg than with ibuprofen 2400 mg. Treatment with etoricoxib reduced the incidence of investigator-reported and confirmed adverse upper GI events by approximately 50% compared with treatment with nonselective NSAIDs. PMID- 12907327 TI - Incomplete response to endoscopic sphincterotomy in patients with sphincter of Oddi dysfunction: evidence for a chronic pain disorder. AB - OBJECTIVES: The efficacy of endoscopic treatment of sphincter of Oddi dysfunction (SOD) with endoscopic sphincterotomy (ES) remains controversial. Although some studies have shown a positive impact on patient symptoms after treatment, these reports have been largely qualitative and evaluated on short-term response. The aim of our study was to quantitatively measure the long-term outcomes of endoscopic therapy in patients with SOD. METHODS: Thirty-three patients with suspected SOD underwent selective sphincter of Oddi manometry (SOM) of the biliary and/or pancreatic sphincter. Each patient completed a telephone-based survey measuring symptomatic pain before and after SOM +/- ES. The questioner was blinded to the results of SOM. The patients with normal SOM or SOD but who did not undergo ES served as controls. RESULTS: Of these 33 patients (27 women, mean age 48.7 yr, range 13-74), 19 (57.5%) were found to have SOD (12 biliary, six pancreatic, one both). The average follow-up was 18.1 months (range 7-34). Of the patients with SOD, 17 (89%) underwent ES. At follow-up of the 19 patients undergoing ES, five were taking narcotics for persistent pain, two were taking antidepressants, and 15 identified the endoscopic therapy as the reason for their relief. Of the 14 controls, seven were taking narcotics, seven were taking antidepressants, and two identified the endoscopy as the reason for their relief; some patients were taking both antidepressants and narcotics. CONCLUSIONS: Patients found to have SOD who undergo ES are more likely to be improved on long term follow-up when compared with patients with suspected SOD but normal manometry without ES. However, almost uniformly, despite ES, patients continue to have pain, which is consistent with most chronic pain disorders and which suggests a multifactorial cause for the pain. PMID- 12907328 TI - Nurse-administered propofol sedation without anesthesia specialists in 9152 endoscopic cases in an ambulatory surgery center. AB - Narcotics and benzodiazepines are commonly used for sedation for endoscopy in the United States. Propofol has certain advantages over narcotics and benzodiazepines, but its use is often controlled by anesthesia specialists. This report describes our experience with dosage, safety, patient satisfaction, and discharge time with nurse-administered propofol sedation in 9152 endoscopic cases. The study was performed in a private practice ambulatory surgery center in Medford, Oregon. With the assistance of an anesthesiologist, we developed a protocol for administration of propofol in routine endoscopic cases, in which propofol was given by registered nurses under the supervision of endoscopists or gastroenterologists. We then applied the protocol with 9152 patients. There were seven cases of respiratory compromise (three prolonged apnea, three laryngospasm, one aspiration requiring hospitalization), all associated with upper endoscopy. Five patients required mask ventilation, but none required endotracheal intubation. There were seven colonic perforations (<1 per 1000 colonoscopies), of which three may have involved forceful sigmoid disruption. Of patients who had previously received narcotic or benzodiazepine sedation, 84% preferred propofol. Gastroenterologists strongly preferred propofol. The mean time from completion of procedures to discharge in a sample of 100 patients was 18 min.Nurse-administered propofol sedation in an ambulatory surgery center was safe and resulted in high levels of patient satisfaction and rapid postprocedure recovery and discharge. PMID- 12907329 TI - Safety of propofol for conscious sedation during endoscopic procedures in high risk patients-a prospective, controlled study. AB - OBJECTIVES: Propofol, a rapidly-acting hypnotic agent, is increasingly being used for endoscopic sedation. Serious adverse effects, including respiratory and cardiovascular depression, make many endoscopists reluctant to use propofol in critically ill patients. This study characterizes propofol's safety profile in consecutive high-risk patients (American Society of Anesthesiologists [ASA] classes III and IV) compared with matched subjects (ASA classes I and II). METHODS: During a 19-month period, 1370 at risk-patients were sedated with propofol, of whom 47% (614 ASA III, 28 ASA IV) were age matched with 642 consecutive patients of the same gender and age assigned to ASA classes I and II and undergoing the same endoscopic procedures (395 gastroscopies, 201 colonoscopies, 14 combined). Registered nurses performed all sedations by propofol dose titration while carefully monitoring arterial oxygen saturation, heart rate, and blood pressure. RESULTS: No major complications occurred among the critically ill patients. There was, however, an increased risk for a short relevant oxygen desaturation (<90%) of 3.6% for ASA III and IV versus 1.7% for ASA I and II (p = 0.036). In four versus one case, short mask ventilation was necessary. Also, a greater proportion of patients showed a > or =5% oxygen saturation decrease. There was no pronounced influence on arterial pressure or heart rate and no perforations in 336 colonoscopies. CONCLUSIONS: With careful monitoring, propofol sedation during GI endoscopies is safe, even for high-risk patients. Considering their higher comorbidity and tendency toward oxygen desaturation, they need particularly careful monitoring, and the required dose is, on mean, 10-20% lower than in ASA classes I and II. PMID- 12907330 TI - Grading portal gastropathy: validation of a gastropathy scoring system. AB - OBJECTIVES: The specific aim of this study was to independently evaluate the reproducibility and validity of the Baveno portal hypertensive gastropathy (PHG) grading system. METHODS: This is a prospective study of 100 consecutive patients with cirrhosis and portal gastropathy. An endoscopy was performed at entry on all subjects and on 70 patients during follow-up (median 15 months). Interobserver reproducibility was assessed by the kappa statistic. Patients with PHG-related bleeding were compared with those without bleeding. The relationship of PHG related bleeding to the PHG score was assessed by a receiver operating characteristic curve and by multiple logistic regression analysis. RESULTS: Interobserver agreement about the presence of the mosaic pattern and red marks was high (kappa >0.75). However, the agreement about the extent of the lesion was poorer (kappa = 0.4-0.45). There was a stepwise increase in PHG-related bleeding risk with increasing PHG scores. Multiple logistic regression confirmed that the PHG score independently correlated with PHG-associated bleeding (OR = 2.5, 90% CI = 1.4-4.6, p < 0.009). During follow-up, red marks developed de novo in one of 70 patients. The severity of red marks worsened in five of 25 patients, whereas they disappeared in eight of 62 individuals. Nodular lesions in the antrum were found in five subjects. The risks of recurrent bleeding during follow-up were related to severe PHG scores (>4), presence of gastric antral vascular ectasia, and nodular lesions in the antrum. CONCLUSIONS: The PHG scoring system is reproducible and accurately reflects the risks of PHG-related bleeding in patients with cirrhosis. PMID- 12907331 TI - Cost-saving approach to patients on long-term anticoagulation who need endoscopy: a decision analysis. AB - OBJECTIVES: The management strategies used when patients requiring long-term anticoagulation need endoscopic procedures vary considerably. Two commonly used approaches are a "heparin window" strategy in the inpatient setting and, more recently, a "switch to low molecular weight heparin (LMWH)" strategy for elective procedures. The aim of this study was to determine whether an initial diagnostic endoscopy (visualization only) is a cost-effective strategy in these patients. METHODS: Decision analysis was performed for two scenarios using probability estimates from our retrospective study. Scenario 1: Patients with any (urgent and elective) indication for endoscopy while on anticoagulation. A decision tree was made outlining two strategies: 1) a diagnostic endoscopy on full anticoagulation followed by therapeutic endoscopy if needed using standard practice; and 2) standard approach. Scenario 2: Patients requiring elective endoscopy. Here, the decision tree outlined three strategies: 1) initial diagnostic endoscopy on full anticoagulation followed by a therapeutic endoscopy if needed using a "heparin window"; 2) initial diagnostic endoscopy followed by therapeutic endoscopy if needed using "switch to LMWH" strategy; and 3) "direct switch to LMWH strategy." RESULTS: Initial diagnostic endoscopy is the preferred strategy when patients requiring anticoagulation need endoscopy. In scenario 1 (all patients), the diagnostic endoscopy approach will reduce need for hospital stay and save $85,006 per 100 patients when a therapeutic impact is not predictable before endoscopy. Similarly, in scenario 2, an initial diagnostic endoscopy followed by switch to LMWH strategy is the most cost saving. CONCLUSIONS: In anticoagulated patients, an initial diagnostic endoscopy approach on anticoagulation is the most cost saving strategy, when a direct therapeutic impact is not predictable. PMID- 12907332 TI - Intestinal cytokines in children with pervasive developmental disorders. AB - OBJECTIVES: A relationship between autism and gastrointestinal (GI) immune dysregulation has been postulated based on incidence of GI complaints as well as macroscopically observed lymphonodular hyperplasia and microscopically determined enterocolitis in pediatric patients with autism. To evaluate GI immunity, we quantitatively assessed levels of proinflammatory cytokines, interleukin (IL)-6, IL-8, and IL-1beta, produced by intestinal biopsies of children with pervasive developmental disorders. METHODS: Fifteen patients, six with pervasive developmental disorders and nine age-matched controls, presenting for diagnostic colonoscopy were enrolled. Endoscopic biopsies were organ cultured, supernatants were harvested, and IL-6, IL-8, and IL-1beta levels were quantified by ELISA. Tissue histology was evaluated by blinded pathologists. RESULTS: Concentrations of IL-6 from intestinal organ culture supernatants of patients with pervasive developmental disorders (median 318.5 pg/ml, interquartile range 282.0-393.0 pg/ml) when compared with controls (median 436.9 pg/ml, interquartile range 312.6 602.5 pg/ml) were not significantly different (p = 0.0987). Concentrations of IL 8 (median 84,000 pg/ml, interquartile range 16,000-143,000 pg/ml) when compared with controls (median 177,000 pg/ml, interquartile range 114,000-244,000 pg/ml) were not significantly different (p = 0.0707). Concentrations of IL-1beta (median 0.0 pg/ml, interquartile range 0.0-94.7 pg/ml) when compared with controls (median 0.0 pg/ml, interquartile range 0.0-60.2 pg/ml) were not significantly different (p = 0.8826). Tissue histology was nonpathological for all patients. CONCLUSIONS: We have demonstrated no significant difference in production of IL 6, IL-8, and IL-1beta between patients with pervasive developmental disorders and age-matched controls. In general, intestinal levels of IL-6 and IL-8 were lower in patients with pervasive developmental disorders than in age-matched controls. These data fail to support an association between autism and GI inflammation. PMID- 12907333 TI - Prospective assessment of Cleveland Global Quality of Life (CGQL) as a novel marker of quality of life and disease activity in Crohn's disease. AB - OBJECTIVES: The Short Form 36 (SF-36) questionnaire is the most widely accepted measure of quality of life (QOL); however, it is cumbersome to use and requires complicated analysis. The Cleveland Global Quality of Life (CGQL) is a simpler tool, which has been validated in patients with ulcerative colitis undergoing restorative proctocolectomy. This study validates CGQL in patients with Crohn's disease and determines the correlation of QOL measurement systems with disease activity as measured by the Crohn's Disease Activity Index (CDAI). METHODS: All patients with Crohn's disease presenting to this department between September 2001 and February 2002 were prospectively entered into a database, including demographic data, disease manifestations, Vienna classification, age at diagnosis, and duration of Crohn's disease. SF-36, CGQL, and CDAI scores were entered concurrently. Cronbach's alpha was used to assess the internal consistency among the components of the CGQL score. Correlation among various QOL measures was assessed with Spearman's test. Relationships between QOL measures and various baseline factors were assessed by Wilcoxon rank sum test, Student's t test, Kruskal-Wallis test, and Pearson's test, depending on the type and distribution of data. Change from baseline in the CGQL was assessed with the Wilcoxon signed rank test. RESULTS: One hundred seventy-eight (178) patients were entered into the database, and 169 completed QOL and CDAI data. There was a significant correlation between SF-36 and CGQL. CGQL and SF-36 correlated similarly with CDAI. Neither patient age, gender, or smoking history were associated with CGQL. CONCLUSIONS: CGQL correlates with disease activity and is a simple measure of QOL in Crohn's disease. PMID- 12907334 TI - Association between physical activity, fiber intake, and other lifestyle variables and constipation in a study of women. AB - Because the risk factors for constipation in the general population are poorly defined, we examined the association between age, body mass index, and selected lifestyle and dietary factors with the prevalence of constipation in the Nurses' Health Study.A total of 62,036 women, aged 36-61 yr and free of cancer, responded to mailed questionnaires in 1980 and 1982, which assessed bowel movement frequency, dietary, and lifestyle factors. Constipation was defined as two or fewer bowel movements weekly.A total of 3327 women (5.4%) were classified as having constipation. Age and body mass index were inversely associated with constipation (p for trend < 0.0001). In multivariate analysis, women who reported daily physical activity had a lower prevalence of constipation (prevalence ratio [PR] = 0.56, 95% CI = 0.44-0.70). Women in the highest quintile of dietary fiber intake (median intake 20 g/day) were less likely to experience constipation (PR = 0.64, 95% CI = 0.57-0.73) than women in the lowest quintile (median intake 7 g/day). The PR for constipation for women who both reported daily physical activity and were in the highest quintile of fiber intake was 0.32 (95% CI = 0.20 0.54) compared with those with physical activity less than once weekly and those who were in the lowest quintile of fiber intake. Higher frequency of aspirin intake was associated with an increasing prevalence of constipation (PR = 1.38, 95% CI = 1.17-1.62). Current smoking and alcohol use were inversely associated with constipation. Our data suggest that moderate physical activity and increasing fiber intake are associated with substantial reduction in the prevalence of constipation in women. PMID- 12907335 TI - The association between corticosteroid use and development of fractures among IBD patients in a population-based database. AB - OBJECTIVES: Because the rate of fracture among patients with inflammatory bowel disease (IBD) is only slightly higher than that in the general population, it is important to define high-risk groups worthy of diagnostic evaluation or prophylactic interventions. Corticosteroid use has been considered in other diseases to be a risk for fracture, although not all studies in IBD are concordant on this point. We aimed to determine whether patients with IBD drawn from a population-based database who sustain fractures are more likely to have been using corticosteroids than a matched group of IBD patients who did not fracture. METHODS: We extracted from our population-based University of Manitoba Inflammatory Bowel Disease Epidemiology Database the number of patients with a new diagnosis of fracture between the years 1997-2000. From within our Inflammatory Bowel Disease Epidemiology Database, we extracted a control group of IBD patients who did not develop fractures matched to the case group who did by age, gender, diagnosis, year of diagnosis, and geographic area of residence. We linked our cohorts with Manitoba Health's Drug Program Information Network to study corticosteroid use within 2 yr before fracture diagnosis. The Drug Program Information Network is a population-based database, established in 1995, which records all prescription drugs. RESULTS: Fractures were identified in 13 patients with Crohn's disease and in 28 patients with ulcerative colitis. The control group included 103 Crohn's disease and 173 ulcerative colitis patients who did not fracture. In Crohn's disease, for the group who fractured compared with the controls who did not fracture, corticosteroid use before fracture was evident in seven (54%) compared with 21 (22%) who did not fracture (chi(2) = 4.45, df = 1, p = 0.035). In ulcerative colitis, for the group who fractured compared with the controls who did not fracture, corticosteroid use before fracture was evident in five (18%) compared with 37 (21%) who did not fracture (chi(2) = 0.031, df = 1, p = 0.861). Fracture cases were more likely to be exposed to oral corticosteroids (OR = 1.75; 95% CI = 0.82-3.75), but this result is not significant. Regarding corticosteroid dosing among the 12 patients with IBD who fractured and used corticosteroids, the mean total days supply was 314 days +/- 236 days compared with 258 days +/- 278 days in those who did not fracture (p = 0.16). The prescribed daily dose among corticosteroid users was comparable for those who fractured versus those who did not fracture (18 mg/day vs 21 mg/day, p = 0.90). CONCLUSIONS: Patients who require corticosteroids in Crohn's disease should be considered at risk for fracture. Further research is required to delineate after how much corticosteroid use are subjects at risk and/or after what duration of active disease. PMID- 12907336 TI - Improved validation of the Inflammatory Bowel Disease Questionnaire and development of a short form in Greek patients. AB - OBJECTIVES: The Inflammatory Bowel Disease Questionnaire (IBDQ) is a disease specific questionnaire for assessing health-related quality of life. The main objective of this study is to assess three main areas of the validation of the Greek IBDQ: internal validity, designating items to dimensions, and selecting items for a short form. An additional aim is to present alternative validation methods for improved and robust results. METHODS: A total of 134 patients with IBD, including 74 with ulcerative colitis (UC) and 60 with Crohn's disease (CD), completed the Greek IBDQ. Internal validity was assessed by using standard methods and multiple correlation analysis. Factor analysis of the 32 items of the questionnaire was carried out to identify the underlying dimensions, using principal factor analysis and maximal likelihood. For selecting items for a short form, three methods of linear regression analysis were used. RESULTS: All items had adequate convergent validity (except item 11 in the CD group) and acceptable discriminant validity (except item 11 for both groups). Factor analysis yielded five factors (emotional function, social function, systemic symptoms, bowel symptoms, and abdominal symptoms). The combination of three regression techniques and their internal validity yielded two similar but distinct short forms, one for UC and one for CD. Both forms had high correlation with the total IBDQ score (R(2) = 0.949 and 0.977, respectively). CONCLUSIONS: The Greek IBDQ had good internal validity. The original designation in four dimensions and the original short form could be improved with only slight modifications. PMID- 12907337 TI - Expression and enzymatic activity of small intestinal tissue transglutaminase in celiac disease. AB - OBJECTIVES: The molecular and functional properties of small intestinal tissue transglutaminase are largely unknown despite growing interest because of its role in celiac disease (CD). In this study, we aimed to evaluate tissue transglutaminase expression and enzymatic activity in bioptic fragments obtained from the duodenum of untreated individuals with CD and from control subjects. METHODS: Analysis of tissue transglutaminase mRNA expression was performed by reverse transcription-polymerase chain reaction (RT-PCR). The presence of the enzyme in bioptic fragments as well as in homogenates from CD patients and controls was revealed by immunohistochemistry and Western blot, respectively, using the antitissue transglutaminase CUB 7402 clone. To evaluate in situ transglutaminase activity, sections of bioptic fragments were incubated in the presence of 5 mmol/L CaCl(2) with 5-(biotinamido)pentylamine or, alternatively, with a biotinylated glutamine-containing hexapeptide (TVQQEL) and the biotinylated 31-43 A-gliadin-derived peptide. RESULTS: Tissue transglutaminase mRNA levels were 1.0-fold higher (p < 0.05) in CD patients than in controls. Immunohistochemistry and in situ demonstration of enzymatic activity in celiac mucosa clearly showed an increased expression of active tissue transglutaminase in the extracellular matrix of the subepithelial region and in the enterocytes. Staining of the biotinylated 31-43 A-gliadin peptide in the same area of tissue transglutaminase suggested the presence of lysine-donor substrates in intestinal mucosa. CONCLUSIONS: Tissue transglutaminase is more expressed and active in defined areas of the small intestinal mucosa from patients with CD. The presence in the celiac mucosa of proteins able to act as amine-donor substrates suggests that tissue transglutaminase-mediated post-translational modification of proteins cross-linked with gliadin peptides may represent a pathogenic mechanism of CD. PMID- 12907338 TI - Infliximab for treatment of pyoderma gangrenosum associated with inflammatory bowel disease. AB - OBJECTIVES: Pyoderma gangrenosum is an immune-mediated inflammatory condition characterized by ulcerative skin lesions affecting 1-2% of patients with inflammatory bowel disease (IBD). Treatment includes wound care, antibiotics, corticosteroids, and immunomodulators. However, response to therapy varies, and many patients with pyoderma gangrenosum have disease that is refractory to these agents. The aim of this study was to assess the response of medically refractory pyoderma gangrenosum to infliximab. METHODS: This was a multicenter retrospective study of patients with IBD and medically refractory pyoderma gangrenosum treated with infliximab. Data collected included the following: baseline demographics; duration of IBD; history of bowel resection; duration of skin lesions; number, size, and location of pyoderma gangrenosum lesions; prior medications; dose and number of infliximab infusions; bowel activity before and after infliximab; pyoderma gangrenosum activity before and after infliximab therapy; time to response and time to healing of pyoderma gangrenosum lesions; recurrence of pyoderma gangrenosum after infliximab; corticosteroid taper; and adverse reactions to infliximab. RESULTS: There were 13 patients with moderate to severe pyoderma gangrenosum and IBD treated with infliximab. All patients demonstrated complete healing of the skin lesions. Three patients had a complete response to induction infliximab therapy and did not require additional treatment. Ten patients responded to induction infliximab and have maintained pyoderma gangrenosum healing with infusions every 4-12 wk. All patients receiving corticosteroids were able to discontinue them completely after institution of infliximab treatment. Infliximab was well tolerated; the only treatment-related adverse events were sunburn in one patient and an infusion reaction in another. CONCLUSIONS: Infliximab is a safe and effective treatment for IBD-associated pyoderma gangrenosum. PMID- 12907339 TI - Chromoscopy might improve diagnostic accuracy in cancer surveillance for ulcerative colitis. AB - OBJECTIVES: Multiple biopsy has been a recommended procedure for cancer surveillance in patients with ulcerative colitis (UC). The aim of this study was to investigate the accuracy of chromoscopic findings in surveillance for patients with UC. METHODS: During the period 1995-2002, we performed 117 surveillance colonoscopies in 57 patients with pancolitis for more than 5 yr. Multiple biopsy specimens were uniformly obtained from flat mucosa in each segment of the colorectum, and, when necessary, from areas specified by chromoscopy. The specified area was classified as polypoid lesion or visible flat lesion. In each specimen, histology was graded according to the classification of dysplasia. RESULTS: Among 818 specimens, 28 (3.4%) were positive for dysplasia. There were 20 low grade dysplasias and eight high grade dysplasias. Dysplasia was more frequently positive in visible flat lesions (37.1%, p < 0001) and in polypoid lesions (16.9%, p < 0.0001) than in flat mucosa (0.4%, p < 0.0001). Furthermore, it was more frequently positive in visible flat lesions than in polypoid lesions (p < 0.05). High-grade dysplasia was found in 4.4% of polypoid lesions and in 14.8% of visible flat lesions, but it was not detected in flat mucosa. Overall, dysplasia was detected in 12 patients. Positive dysplasia was confined to visible flat lesions in four patients and to flat mucosa in one patient. CONCLUSIONS: Our results suggest that biopsy from flat visible lesions under chromoscopy might improve the accuracy of cancer surveillance in UC. PMID- 12907340 TI - Colonic ulcers accompanying collagenous colitis: implication of nonsteroidal anti inflammatory drugs. AB - OBJECTIVES: A small minority of otherwise typical collagenous colitis (CC) patients also have mucosal ulceration (CC-U). We studied the association of CC-U cases with ingestion of nonsteroidal anti-inflammatory drugs (NSAIDs) as a possible explanation for the mucosal ulceration. METHODS: Clinical information and histological features were reviewed in nine cases of biopsy-diagnosed CC-U. Biopsies from 18 unselected cases of CC without ulceration were reviewed for comparison. RESULTS: Of nine patients with CC-U, seven (77.8%) had a history of NSAID ingestion, compared with four of 18 CC controls (20.2%) (p = 0.006). The diarrhea resolved after cessation of NSAID use in four CC-U patients, partially resolved in one patient, and persisted in one patient. The outcome was not available in one patient. Of the two CC-U patients who did not use NSAIDs, one patient was taking lisinopril (angiotensin-converting enzyme inhibitor), and the diarrhea resolved after stopping the drug; the ulceration in the second patient was thought to be ischemic in origin. CONCLUSIONS: Collagenous colitis with ulceration has a strong association with NSAID ingestion. Evaluation of medications and cessation of NSAIDs should be considered as a therapeutic option in cases of collagenous colitis with colonic ulceration. PMID- 12907341 TI - Association between pain episodes and high amplitude propagated pressure waves in patients with irritable bowel syndrome. AB - OBJECTIVES: In the pathogenesis of irritable bowel syndrome (IBS), both increased visceral sensitivity and altered colonic motility seem to play a role. The aim of this study was to quantify the temporal relationship between pain episodes and the occurrence of high amplitude propagated pressure waves (HAPPWs). METHODS: A total of 11 IBS patients with the nonconstipation predominant pattern of IBS and 10 sex- and age-matched healthy volunteers were studied. On day 1, a solid state manometric catheter was positioned in the left colon and connected to a data logger. The subjects then went home. Thereafter they pressed a button on the data logger at the beginning and end of each pain episode. The 24-h manometric signal recorded on day 2 was divided into consecutive 5-min periods. These periods were evaluated for the occurrence of pain and HAPPWs. Fisher's exact test was applied to calculate the probability that HAPPWs and pain episodes were unrelated. The symptom association probability (SAP) was calculated as (1.0 - P) x 100%. A SAP of >95% was considered to indicate that the observed association did not occur by chance. RESULTS: In four of seven patients with pain on day 2, the SAP was >95%. HAPPWs that were related to pain originated at a more proximal level (p = 0.026) and occurred earlier (p = 0.007) than HAPPWs that were not related to pain. The duration of a pain period was correlated with the number of pain-related HAPPWs in that period (r = 0.906, p = 0.013). Two of the 10 healthy subjects experienced pain, and these pain episodes were not associated with HAPPWs. CONCLUSIONS: Using objective analysis techniques, an association between pain episodes and HAPPWs was found in nonconstipated IBS patients with pain. HAPPWs that are associated with pain are only slightly different from HAPPWs not associated with pain. PMID- 12907342 TI - Validation of automated blood cell counter for the determination of polymorphonuclear cell count in the ascitic fluid of cirrhotic patients with or without spontaneous bacterial peritonitis. AB - OBJECTIVES: Polymorphonuclear (PMN) cell count in ascitic fluid is the most useful test for the diagnosis of spontaneous bacterial peritonitis (SBP). We evaluated the validity of an automated blood cell counter for the PMN determination in ascitic fluid by comparing it with the traditional hematologic method with a light microscope in a manual counting chamber. METHODS: A total of 130 ascitic fluid samples were collected from 74 consecutive cirrhotics. The agreement between the two techniques was assessed according to Bland and Altman's method. The sensitivity, specificity, and positive and negative predictive values of the automated blood cell counter were calculated by considering the diagnosis of SBP as a PMN count > or = 250 cells/mm(3), determined by the manual method as the "gold standard." RESULTS: The mean PMN counts assessed by the manual method and the automated blood cell counter were 124 +/- 301 cells/mm(3) and 130 +/- 339 cells/mm(3), respectively (p = 0.89, ns). The mean +/- SD of the difference between manual and automated measurements was 6 +/- 61 cells/mm(3), whereas the limits of agreement were +127 cells/mm(3) (95% CI = +108 to +147) and -115 cells/mm(3) (95%CI = -96 to -135). SBP was diagnosed in 11 patients. All but one were correctly identified with the automated blood cell counter, with a sensitivity of 94% and a specificity of 100%; positive and negative predictive values were 100% and 99.1%, respectively. CONCLUSIONS: The manual method and the automated blood cell counter have a good agreement in the PMN determination in ascitic fluid, and the automated blood cell counter is a reliable tool for rapid diagnosis of SBP. PMID- 12907343 TI - Metabolic and anthropometric evaluation of insulin resistance in nondiabetic patients with nonalcoholic steatohepatitis. AB - OBJECTIVES: Insulin resistance is nearly universal in patients with nonalcoholic steatohepatitis (NASH) when tested by glucose tolerance tests or clamp methods. However, the pattern of insulin resistance in these patients after a physiological challenge is unknown. We conducted a study to characterize the metabolic response to a mixed meal in nondiabetic patients with NASH (NDN) and to identify anthropometric determinants of insulin resistance in these patients. METHODS: Serum insulin, C-peptide, glucose, and free fatty acid (FFA) levels were measured at 0, 30, 60, 90, and 120 min after a 500-kcal standard meal in 18 NDN and 18 age-, gender-, and body mass index (BMI)-matched controls. Correlations were made between insulin resistance and various anthropometric, calorimetric, and serological variables. RESULTS: Compared with controls, NDN had significantly higher levels of insulin and C-peptide at baseline and after the mixed meal. However, glucose levels were not different either at baseline or after the meal. NDN had higher fasting levels of FFA than the controls (459 +/- 190 vs 339 +/- 144 micro mol/L, respectively, p = 0.03); however, meal-induced suppression in lipolysis was similar between the two groups (39 +/- 113% vs 46 +/- 60%, p = 0.8). Insulin resistance was significantly correlated with BMI (r = 0.39, p = 0.02) and visceral fat (r = 0.50, p = 0.004). Whereas BMI, percent total body fat, and subcutaneous abdominal fat were similar between the groups, the NASH group had significantly higher percent visceral fat compared with controls (28 +/ 10% vs 22 +/- 14%, p = 0.02). CONCLUSIONS: NDN are significantly hyperinsulinemic, both at fasting and after the mixed meal; however, their glucose homeostasis and suppression in lipolysis after a meal challenge are maintained. Insulin resistance in these patients is likely related to their higher visceral fat mass. PMID- 12907344 TI - Risk factors for symptomatic gallstones in patients with liver cirrhosis: a case control study. AB - OBJECTIVES: Liver cirrhosis is a well-documented risk factor for the formation of gallstones. In cirrhotic patients, gallstones are almost always "silent," and surgery is rarely required. When indicated (symptoms or complications), cholecystectomy implies a high morbidity risk in these patients, especially in the advanced stages of cirrhosis. The aim of this study was to estimate the risk factors for symptom development in cirrhotic patients with gallstones to identify the subgroup of patients at risk of undergoing surgery. METHODS: A total of 140 patients with liver cirrhosis and gallstones were studied: 97 with asymptomatic and 43 with symptomatic gallstone disease. The risk factors for gallstone formation (age, gender, family history, parity, obesity, diabetes mellitus, hyperlipoproteinemia) and the characteristics of liver cirrhosis (etiology, duration, Child class, hypersplenism), gallstones (duration, number, size), and gallbladder (size, wall thickness) were assessed in all patients. In 12 patients (four symptomatic, eight asymptomatic), gallbladder emptying was also evaluated by ultrasound. The association of asymptomatic and symptomatic gallstones with all these parameters was statistically evaluated by Student's t, Mann-Whitney, and chi(2) tests, as well as by means of multiple logistic regression. The causal relationship between these characteristics and gallstone symptoms was also examined by means of the KDD (knowledge discovery from databases) method, with an algorithm for learning Bayesian networks. RESULTS: Advanced age, female gender, viral etiology of cirrhosis, family history of gallstones, and duration of gallstone disease were significantly associated with symptomatic gallstone disease. The number or size of gallstones and the size or emptying of the gallbladder did not differ in symptomatic versus asymptomatic patients. Male gender and alcoholic cirrhosis were inversely correlated with symptom presence. In the multivariate analysis, family history (p = 0.0098) and advanced age (p = 0.0422) were positively correlated and male gender (p = 0.0049) and alcoholic etiology of cirrhosis (p = 0.0116) negatively correlated with symptom presence. These relationships (except for age) were also evidenced by the KDD method. CONCLUSIONS: The risk of gallstones becoming symptomatic is significantly lower in men and in alcoholic cirrhosis. In cirrhotic women, and especially in the presence of a positive family history and of advanced age, the risk of developing symptoms and undergoing surgery was significantly greater. PMID- 12907345 TI - Biliary casts after orthotopic liver transplantation: clinical factors, treatment, biochemical analysis. AB - OBJECTIVES: Biliary casts develop in up to 18% of liver transplant recipients. Casts are associated with morbidity, graft failure, need for retransplantation, and mortality. Proposed etiological mechanisms include acute cellular rejection, ischemia, infection, and biliary obstruction. We aimed to identify clinical features associated with biliary cast formation, review treatments, and analyze the biochemical composition of casts at a single, large, liver transplant center. METHODS: Patient records were reviewed retrospectively to identify patients who developed casts. Data were collected with attention to ischemia, rejection, obstruction, infection, immunosuppression, postoperative biliary drain use, and cast-directed management, and were compared with data from controls. Cast specimens, retrieved at cholangiography, were analyzed with chromatography techniques. RESULTS: Ischemic factors were noted in 70% (7/10) of cast patients versus 15% (6/40) of controls (OR = 13.2; 95% CI = 2.7-66.0; p = 0.001). Biliary strictures were present in 50% of cast patients versus 10% of controls (OR = 9.0; 95% CI = 1.8-45.2; p = 0.01). Differences in cold ischemia time, acute cellular rejection, cyclosporin use, infection, and postoperative biliary drain use were not significant. Casts were successfully treated by endoscopic and percutaneous methods in 60% of patients. One patient died of cast-related complications (mortality 10%). Four casts were in satisfactory condition for biochemical analysis. Bilirubin was the main component ( approximately 10-50%). Bile acid synthesis products and cholesterol comprised smaller percentages, and protein comprised only 5-10%. CONCLUSIONS: Biliary casts are more likely to develop in the setting of hepatic ischemia and biliary strictures. Endoscopic and percutaneous cast extraction might achieve favorable results and should be attempted before surgical therapy. PMID- 12907346 TI - Desmoid tumors in familial adenomatous polyposis: a pilot project evaluating efficacy of treatment with pirfenidone. AB - OBJECTIVES: Pirfenidone (Deskar, Marnac Inc., Dallas, TX), 5-methyl-1-phenyl-2 (1H)-pyridone, is a broad-spectrum, noncytotoxic, oral antifibrotic agent that is reported to inhibit or block the action of cytokine growth factors: transforming growth factor beta1, platelet-derived growth factor, epidermal growth factor, and fibroblast growth factor, and to prevent formation of new fibrotic lesions. METHODS: We enrolled 10 women and four men with extensive familial adenomatous polyposis (FAP)-associated desmoid disease in a 2-yr open-label treatment trial with oral pirfenidone. Imaging of desmoids was conducted at baseline and 6, 12, and 24 months. RESULTS: No drug toxicity or drug intolerance was encountered. Seven patients dropped out (three because of progressive disease), and seven continued for at least 18 months. Of those that continued, two had partial but significant reduction in the size of all desmoids beginning in the first 6 months of treatment, and two others experienced relief of symptoms without change in desmoid size. Three patients experienced no change in tumor size or symptoms. CONCLUSIONS: Pirfenidone is well tolerated by patients with FAP-associated desmoid tumors. Some patients with FAP/desmoid tumors treated with pirfenidone had regression of tumors, some had progression, and some had no response. Patients with rapidly growing tumors did not respond to pirfenidone. A placebo controlled trial is needed to determine whether there is a subset of patients for whom pirfenidone may result in partial shrinkage of desmoid tumors, because the natural history of desmoid tumors is not predictable or understood. PMID- 12907347 TI - Determinants of colorectal cancer screening in women undergoing mammography. AB - OBJECTIVES: Women who participate in screening for breast cancer are more likely to participate in screening for colorectal cancer. We studied such a motivated group of women to identify predictors of, and barriers to, participation in colorectal cancer screening by endoscopy. METHODS: We distributed surveys to 551 women > or = 50 yr of age while they were awaiting mammography at four sites in and around Boston, MA from June to September, 2000. The 40-question survey assessed knowledge, attitudes, and beliefs about, and behaviors toward, breast and colorectal cancer screening. Regression models were used to determine factors associated with having had sigmoidoscopy or colonoscopy. RESULTS: Seventy-nine percent of the women completed all or part of the survey. Half (221/438) reported ever having had sigmoidoscopy or colonoscopy. Of these, 93% did so at the recommendation of their primary care provider. Factors associated with participation in endoscopic screening included compliance with annual fecal occult blood testing, a family history of colorectal cancer, and indifference toward the gender of the doctor performing the endoscopy. CONCLUSIONS: Women undergoing mammography overwhelmingly cite the recommendation of their primary care provider as the reason for participating in colorectal cancer screening by endoscopy. Women who preferred a female endoscopist were less likely to have been screened. Whenever possible, primary care providers should offer women the choice of a female endoscopist for colorectal cancer screening. PMID- 12907348 TI - Colonoscopy screening in the elderly: when to stop? AB - OBJECTIVES: The age to begin colorectal cancer (CRC) screening is based on the risk of neoplasia and is published in screening guidelines. The age to stop screening is unknown but should be based, in part, on the same principle. The purpose of this study was to establish whether the prevalence of neoplasia detected by colonoscopy diminished with advancing age, to warrant ceasing colonoscopic screening. METHODS: The endoscopic and pathology reports of all asymptomatic subjects undergoing colonoscopy for the purpose of CRC screening or an evaluation of abdominal pain or change in bowel habits between 1997 and 2000 were reviewed. A multivariate logistic regression analysis was used to assess the effect of age, gender, and indication for examination on the prevalence of neoplasia, as well as on having more than two adenomas, advanced adenomas (tubulovillous, villous, severe dysplasia, or size > or = 1 cm), and invasive cancers. RESULTS: A total of 915 patients were included. Of these, 50% were male, with a mean age of 65 yr (range 50-100). Neoplasia peaked in the seventh decade, with a fall thereafter (p = 0.009). Numerous adenomas, advanced adenomas, and invasive cancers increased with age. The yield for overall neoplasia, advanced adenomas, and more than two adenomas was higher in the screening group than in the symptomatic group. More invasive cancers were found in the symptomatic group compared with the asymptomatic group, but this did not achieve statistical significance (4 vs 1, p = 0.44). CONCLUSIONS: The prevalence of advanced neoplasia continues to increase with age. Subjects undergoing colonoscopy for screening had a greater risk for neoplasia than did subjects with symptoms. There is no decline in yield of advanced neoplasia to justify stopping screening colonoscopy in the elderly. PMID- 12907349 TI - Band ligation versus propanolol and isosorbide mononitrate for primary prophylaxis of variceal bleeding. PMID- 12907350 TI - Hepatitis B-lamivudine for all and forever?. PMID- 12907351 TI - Outcomes of treatment in achalasia: subjective or objective endpoints?. PMID- 12907352 TI - Re: Johnson et al. endoscopic, deep mural implantation of enteryx for the treatment of GERD: 6-month follow-up of a multicenter trial. PMID- 12907354 TI - Bleeding due to repeat sphincterotomy in the early period. PMID- 12907356 TI - Hospitalization and mortality rates from peptic ulcer in the 1990s. PMID- 12907358 TI - Mucinous ductal ectasia: making the numbers really count. PMID- 12907359 TI - Georg Ernst Konjetzny, German surgeon of the 20th century: a great pioneer who suggested the bacterial genesis of gastritis and its relationship to peptic ulcer and gastric cancer. PMID- 12907360 TI - Genetic fingerprinting helps to resolve an unpleasant mistake. PMID- 12907361 TI - Comparison of three integral tight junction barrier proteins in Barrett's epithelium versus normal esophageal epithelium. PMID- 12907362 TI - Percutaneous endoscopic gastrostomy in patients with dementia. PMID- 12907365 TI - Detection of extracellular forms of babesia in the blood by electron microscopy: a diagnostic method for differentiation from Plasmodium falciparum. AB - Light microscopic examination of blood smears is the traditional approach for the diagnosis of babesiosis, but there is morphological overlap with Plasmodium falciparum. The authors describe 3 patients with babesial infection in whom the diagnosis was made by identifying extracellular merozoites in a buffy coat preparation using electron microscopy. This method resulted in a high yield of extracellular babesia, even in a case where virtually no extracellular babesia were detectable in the blood smear. The test had a reasonably fast turnaround time and allowed detailed visualization of the organisms and reliable distinction from Plasmodium falciparum. PMID- 12907366 TI - Molecular variability of TLS-CHOP structure shows no significant impact on the level of adipogenesis: a comparative ultrastructural and RT-PCR analysis of 14 cases of myxoid/round cell liposarcomas. AB - A specific TLS-CHOP fusion gene derived from the t(12;16) is present in at least 95% of myxoid/round cell liposarcomas (MLS). Rare cases of MLS show a variant t(12;22) translocation, resulting in EWS-CHOP fusion gene. The CHOP gene encodes a leucine-zipper transcription factor, which is implicated in both oncogenic transformation and inhibition of adipogenesis. To examine whether the molecular variability of TLS-CHOP or EWS-CHOP fusion transcript structure is associated with the degree of inhibition of adipogenesis, a comparative ultrastructural and RT-PCR-based analysis of 14 MLS was performed. The specimens consisted of 9 primary, 2 locally recurrent tumors, and one representative sample each from 3 patients with multifocal soft tissue metastases. Histologically, there were 8 high-grade and 6 low-grade MLS using 5% round cell (RC) component as the cutoff point. By RT-PCR assay there were 13 cases with TLS-CHOP fusion transcripts: 7 cases of type 5-2 (known as type II), 4 cases of type 7-2 (known as type I),1 case of type 8-2 (known as type III), and 1 unique case of type 6-2. The remaining 1 case showed an EWS-CHOP fusion transcript. Ultrastructural examination revealed that tumor cells were composed of a moderate-to-predominant proportion of well-formed lipoblasts in 4 cases, while in 6 cases such lipoblasts were very scant. The remaining 4 tumors were arrested in the stage of transitional cells. The heterogeneity of TLS-CHOP fusion transcript showed no apparent impact on adipogenesis, since both TLS-CHOP type I and II cases could randomly display various levels of lipoblastic differentiation. Furthermore, the 4 cases without definite lipoblasts showed no preference for any specific fusion variants and consisted of one each of TLS-CHOP subtypes. In addition, the fusion transcript variants did not correlate with other ultrastructural features, such as the presence and amount of glycogen, mitochondria, rough endoplasmic reticulum, vimentin-like intermediate filaments, and external lamina. However, there appeared to have a trend suggesting the predilections of glycogen particles and vimentin-like intermediate filaments in primitive mesenchymal cells and/or transitional cells. These findings cannot substantiate the hypothesis that the molecular variability of fusion transcripts has a biological impact on adipogenesis of MLS, and other factors might be implicated in their level of differentiation. PMID- 12907367 TI - Aggressive angiomyxoma: an ultrastructural study of four cases. AB - The histogenesis of the aggressive angiomyxoma of the vulvo-vaginal region was studied. Four cases of aggressive angiomyxoma were examined by using light microscopy, electron microscopy, and immunohistochemistry. Myofibroblastic from smooth muscle cell differentiation was distinguished by paying close attention to the structures of the neoplastic cell membrane. Aggressive angiomyxomas exhibit subtle features of smooth muscle differentiation, suggesting that the neoplastic cells differentiate from a multipotent perivascular cell. PMID- 12907368 TI - An ultrastructural, immunohistochemical, and cytogenetical study of a monophasic pulmonary synovial sarcoma: implications of the frequent ultrastructure of oligocilia and concentric membranous bodies with positive immunostaining for VS38c and MEF2. AB - A rare case of a monophasic pulmonary synovial sarcoma is reported. A 44-year-old Japanese man underwent lower lobectomy for a nodular mass in his right lung. Immunohistochemical study of the excised primitive spindle cell sarcoma revealed occasional positive stains by hitherto reported antigens of S-100, cytokeratin 7, high molecular weight cytokeratin (34 beta E12), pankeratin (AE1/AE3), and EMA, which were helpful for the differential diagnosis of other spindle cell sarcomas. Furthermore, positive immunostains for MEF2, VS38c (plasma cell antigen), and bcl 2 were rather significant findings in the present case. The definitive evidence that molecular genetic analysis showed a clonal single electrophoretic band of SYT-SSX mutated chimera gene was conclusive for the pathological diagnosis. The implications of the frequently seen ultrastructure of oligocilia and concentric membranous bodies with positive stains for VS38c and MEF2 are discussed. In the difficult pathological diagnosis of a rare and undifferentiated type of sarcoma with unusual clinicopathological features generated at an unusual site, comprehensive ultrastructural, immunohistochemical, and cytogenetic studies will lead to the correct pathological diagnosis and elucidate the detailed characteristics of the tumor. PMID- 12907369 TI - Clinico-pathological evaluation of ciliary dyskinesia: diagnostic role of electron microscopy. AB - From November 1995 to May 2002, the authors studied 59 children with suspected primary ciliary dyskinesia (PCD). Samples of ciliated respiratory epithelium were obtained by nasal brushing from 44 patients and by biopsy of bronchial mucosa from 15 patients. Thirty-four/Fifty-nine samples were suitable to obtain a qualitative-quantitative evaluation of ultrastructural ciliary abnormalities. Many ciliary and axoneme alterations were described. This study revealed that quantitative and qualitative ultrastructural assessment of respiratory epithelial cilia plays an important role in the differentiation between primary, secondary, and borderline ciliary abnormalities. Early diagnosis of PCD with appropriate clinical follow-up and treatment is important to prevent irreversible lung tissue damage, namely bronchiectasis. PMID- 12907370 TI - Dose-dependent mercuric chloride tubular injury in rat kidney. AB - Mercuric chloride (HgCl(2)) produces an acute renal failure in experimental animal models. Since the mechanism of tubular injury has not completely been clarified, this morpho-quantitative study was undertaken to better understand the effects of 2 sublethal doses (T1=1 mg/kg and T3.5=3.5 mg/kg) of HgCl(2) in rat proximal tubules. Morphometrical analysis was performed to quantify both cytoplasmic and nuclear changes found in treated in respect to saline-injected proximal tubules. In the controls, single-cell damage was occasional and nucleolar changes were absent. HgCl(2) induced progressively severe proximal tubule atrophy. In the T1 group, necrosis was limited to pars recta cells and nucleolar segregation was often partial. In the T3.5 group, atrophy was extensive in both convoluted and straight tracts, the nucleolus was completely segregated and coiled body-like inclusions were detached from it. Ultrastructural analysis confirmed dose-dependent changes within straight proximal tubules, i.e., necrosis, apoptosis, nucleolar segregation, swollen mitochondria, vacuolization, and disrupted brush border. In conclusion, in the rat kidney HgCl(2) induced dose dependent alterations not only in the cytoplasm but also in the nucleus of proximal tubule cells. These findings will be useful for better understanding of the pathogenesis of mercury nephrotoxicity and its genotoxic effect. PMID- 12907371 TI - Size of azurophil lysosomes of human PMNs determined by stereological morphometry. AB - The most striking feature of polymorphonuclear leukocytes is the presence of the subcellular organelles known as lysosomes. The peroxidase positive azurophil (primary) lysosomes are generally described in textbooks and the journal literature as predominantly ellipsoidal and approximately 500 nm in size. However, there appears to be no discovery research in the literature establishing that value. This is the first investigation to utilize stereological techniques for microscopic morphometry to more accurately determine the size of human azurophil (primary) lysosomes. This study reports the size of ellipsoidal azurophil lysosomes to be 335 nm with a standard deviation of 97 nm. PMID- 12907372 TI - Rokitansky-Aschoff sinuses of the gallbladder are associated with black pigment gallstone formation: a scanning electron microscopy study. AB - Rokitansky-Aschoff sinuses are the result of hyperplasia and herniation of epithelial cells through the fibromuscular layer of the gallbladder wall and are usually referred to as adenomyomatosis. The role of this study is to demonstrate that Rokitansky-Aschoff sinuses of the gallbladder are a risk factor for the formation of black pigment gallstones. A total of 179 removed gallbladders, were hystologically examined. Sixty-four of the 179 consecutive cholecystomized patients had typical adenomyomatosis. Thirty-eight of the 64 patients with adenomyomatosis had black pigment gallstones, alone (n=22) or in association with single (n=12) or multiple (n=4) cholesterol gallstones in the same gallbladder. Twelve of these patients did not have the typical risk factors for black stones (hemolysis, cirrhoses, gastrectomy, etc). Gallstones were examined by infrared spectroscopy and X-ray diffractometry. In addition, in a subset of 14 patients, the gallstones and the gallbladder wall were examined by scanning electron microscopy. At least in the initial phases of formation, Rokitansky-Aschoff sinuses were found close to small intraparietal vessels and sometimes they contained black pigment microstones. After the fourth to fifth decades of life, black gallstones can be found in the Rokitansky-Aschoff sinuses and in the main gallbladder lumen. Black pigment gallstones can form in Rokitansky-Aschoff sinuses of the gallbladder in absence of the typical risk factors for bilirubin suprasaturation of bile. PMID- 12907373 TI - Identification of Prevotella in pedal osteomyelitis of a diabetic patient. AB - Osteomyelitis in a diabetic patient with a nonhealing foot ulcer, multiple medical conditions, and recurrent hospitalization for antibiotic therapy was found to be associated with gram-negative bacteria Prevotella melanginoganica and Prevotella melaninoganica hemagglutinating variant. Those organisms were identified due to the morphologically distinct features in electron microscopy and sequencing of the genes after Polymerase chain reaction amplification from the pathological material. The bacteria invaded the bone and resided in osteocyte, osteoblast, and endothelial cells. The bacteria are usually associated with periodontal plaques, causing inflammation and destruction of gingival tissue and resorption of the alveolar bone. This is the first ultrastructural and molecular study of a diabetic bone lesion with anaerobic bacterial infection. PMID- 12907376 TI - American Academy of Nursing Expert Panel on Violence policy recommendations for nurses caring for victims of torture (adopted 2001). PMID- 12907377 TI - American Academy of Nursing Expert Panel on Violence policy recommendations on workplace violence (adopted 2002). PMID- 12907378 TI - Therapeutic relationships and boundary maintenance: the perspective of forensic patients enrolled in a treatment program for violent offenders. AB - To extend current knowledge about therapeutic relationships and boundary maintenance with the incarcerated forensic patient, the focus of this naturalistic inquiry was the exploration of the perspectives of forensic patients enrolled in a treatment program for violent offenders. Twelve male participants were interviewed three times. Eight of the twelve participants were interviewed a fourth time for the purpose of soliciting their feedback regarding the researcher's analysis of the data. Analysis of the data collected revealed a core process--the development of "therapeutic" relationships--indicating that the development of relationships was a complex process. Consistent with the participants' tendency to use terms analogous to a house to describe their experiences, the analogy of a house was used to describe the five interrelated themes that emerged from the data. If the interpersonal relationship is the heart of nursing then forensic mental health nurses need to understand the complexity of therapeutic relationships from the perspective of their patients. PMID- 12907379 TI - The tragedy of torture: a global concern for mental health nursing. AB - Torture, a tool of aggressive political regimes, exists in over 100 nations around the world and, as documented in archaeological evidence, has been a part of the dark side of human history, since at least 1500 B.C. In 1864, the first Geneva Convention aimed at protecting wounded soldiers in wartime, and 120 years later, in 1984, the United Nations adopted a specific Convention against Torture. Earlier, in 1975, nurses, through the International Council of Nursing (ICN), adopted a position on caring for people who had been tortured. Danish nurses and physicians have been leaders in advocating the elimination of torture in the world. Yet, nurses in the United States, until the adoption of a resolution by the American Academy of Nurses, October, 2001, had been quite silent about this human pain. With the tragedy of 9/11 still fresh in our minds, nurses are urged to be vigilant in curtailing any torture of prisoners associated with this event. Treatment plans for victims follow many of the interventions used with posttraumatic stress disorder, yet there are unique differences as well. PMID- 12907380 TI - Case law regarding nurses as expert witnesses in child abuse. PMID- 12907381 TI - Positive self-transitions in women child abuse survivors. AB - Clinicians need information about what factors contribute to women child abuse survivors' mental health and success. Most previous research has focused on negative after-effects of abuse, and correlating types and severity of abuse with specific mental and physical problems in adulthood. Two qualitative studies (total N = 55) were done with low-income women survivors of child abuse and neglect from a midwestern inner city area. The majority of these women were African American. All participants were in recovery from substance abuse problems. A secondary analysis of these accounts was done to examine positive life transitions made by these women after experiences of abuse, and throughout adulthood, despite having serious after-effects of abuse. The investigator and research team categorized themes from the narrative data into two processes- epiphanies and maintaining momentum--and six elements constituting the content areas of self-change: self-centering, ownership, interpersonal insulation, willfulness, seeing options, and spiritual connection. Maintaining momentum and shortening the time between epiphanies was found to be central. Tentative clinical implications are discussed. The study was preliminary in that it was a secondary analysis. Suggestions for further research are outlined. PMID- 12907382 TI - Women's perceptions of their male batterers' characteristics and level of violence. AB - This article describes the characteristics of male perpetrators of domestic violence and their relationship to the level of violence. The data about the male partners obtained from 151 battered women were used for this analysis. Using multiple regression, demographic variables and three behavioral indicators, including use of alcohol before a violent episode, history of arrests, and the generality of violence, were examined together for their relationship with the violence scores. With the level of violence as measured by the Conflict Tactics Scale (CTS) as the dependent variable, demographic variables explained 19.1% of the variability, with the behavioral indicators accounting for an additional 4.6% of the variability. Several research and clinical implications are addressed. PMID- 12907383 TI - Strength and vulnerability: spirituality in abused American Muslim women's lives. AB - The importance of spirituality for individuals coping with and recovering from trauma has been widely recognized. Despite this recognition, little information is available addressing the influence of spirituality on the abuse experiences of women surviving intimate partner violence (IPV). This paper begins to amend this gap in knowledge by examining the influence of spirituality on the abuse experiences of American Muslim women, a large and growing population. Findings from this qualitative study indicate that spirituality provided participants with an important means of coping with ongoing violence while in many instances also creating barriers to safety. These findings underscore the complex role spirituality may play as a source of both strength and vulnerability in American Muslim women's response to IPV. PMID- 12907384 TI - Isolation, gratification, justification: offenders' explanations of child molesting. AB - Child sexual abusers' descriptions of their thoughts, behaviors, and relationships were identified by reviewing qualitative research studies published between 1982 and 2001. Offenders used cognitive distortions to meet personal needs, protect themselves from aversive self-awareness, and overcome internal inhibitions against engaging children in sexual activity. Offenders carefully groomed their victims by systematically separating them from their families and peers and socializing them into sexual relationships. Specific cognitive and behavioral processes were used to translate sexual arousal into offending behavior. Offenders justified the abuse by making excuses and redefining their actions as love and mutuality. Throughout the process, offenders exploited the power imbalance inherent in all adult-child relationships. The participants in these studies provided a unique perspective on child sexual abuse that can be used to help victims cope, guide treatment development and evaluation, and suggest new questions for future research. PMID- 12907385 TI - Family dynamics and child abuse and neglect in three Finnish communities. AB - The purpose of this study was to describe family dynamics of Finnish families in which there is abuse or neglect of a child in that family. One adult from 41 families with recognized child abuse/neglect completed the Family Dynamics Measure II (FDM II) consisting of six dimensions of family life, and answered questions about problems, changes and illnesses. Nurses and social workers in community health care and social agencies identified potential subjects in their agencies and collected data. Four of the six dimensions of the FDM II reached statistical significance (p =.05) with child age and number of children, education, marital status, and age of respondent. They are: Flexibility vs. rigidity, Stability vs. disorganization, Clear vs. distorted communication, and Role reciprocity vs. role conflict. FDM II is useful in identifying several of the less positive family dynamics dimensions in abuse:neglect families. Conclusions for clinical management cannot be drawn at this time. PMID- 12907386 TI - The wavering line in the sand: the effects of domestic violence and sexual coercion. AB - Control and power regarding sexuality and fertility challenges many women, especially those involved in abusive relationships. This preliminary study was done to explore the relationship between domestic violence, sexual coercion, and pregnancy. The sample was comprised of community-dwelling women attending support groups for survivors of abuse, and women attending group therapy sessions while housed at a battered woman's shelter. Both questionnaires and focus groups addressed the women's experiences with contraception, sexuality, pregnancy, and domestic violence. Only the qualitative phase of the study, plus demographics from the questionnaires, are presented in this article. The responses suggest that many of the women felt they could not avoid intercourse with their abusers, despite fears of pregnancy. Focus group transcript analysis revealed recurrent themes of problems regarding pregnancy, tactics of abuse, inability to access birth control, and denial and mistrust. This article explores the reality these women face and the thought processes they employ to survive and remain in their relationship. Health care providers can use this information to better understand their clients, to assist them in obtaining and using effective contraception, and to support clients as they make their decisions. PMID- 12907387 TI - Development of a recombinant vaccine against Japanese encephalitis. AB - Japanese encephalitis (JE) is the major form of viral encephalitis in much of the South-East Asia, India, and China. The disease is caused by a mosquito-borne virus known as Japanese encephalitis virus (JEV). The virus spreads in the form of epidemics, although several endemic areas for JEV activity are known. In recent years, JEV has spread to newer geographic locations such as Australia and Pakistan, and thus has become an important emerging virus infection in these areas. A mouse brain-derived, formalin-inactivated vaccine is available for immunization against JE. Because the formalin-inactivated JEV vaccine has limitations in terms of safety, availability, and cost, attempts are being made to develop improved vaccine using the recombinant DNA technology. This article reviews various attempts in this direction and summarizes the latest developments such as the recombinant yellow fever virus- or the plasmid DNA-based JEV vaccine. PMID- 12907388 TI - An unusual syncytia-inducing human immunodeficiency virus type 1 primary isolate from the central nervous system that is restricted to CXCR4, replicates efficiently in macrophages, and induces neuronal apoptosis. AB - Macrophage/microglia cells are the principal targets for human immunodeficiency virus type 1 (HIV-1) in the central nervous system (CNS). Prototype HIV-1 isolates from the CNS are macrophage (M)-tropic, non-syncytia-inducing (NSI), and use CCR5 for entry (R5 strains), but whether syncytia-inducing (SI) CXCR4-using X4 strains might play a role in macrophage/microglia infection and neuronal injury is unknown. To explore the range of features among HIV-1 primary isolates from the CNS, the authors analyzed an HIV-1 strain (TYBE) from cerebrospinal fluid of an individual with acquired immunodeficiency syndrome (AIDS) that was unusual because it was SI. Like other CNS isolates, HIV-1/TYBE replicated to high level in primary human macrophages, but, in contrast to CNS prototypes, TYBE used CXCR4 exclusively to infect macrophages. A functional TYBE env clone confirmed the X4 phenotype and displayed a highly charged V3 sequence typical of X4 strains. Supernatant from TYBE-infected primary human macrophages induced apoptosis of neurons. Thus, TYBE represents a novel type of CNS-derived HIV-1 isolate that is CXCR4-restricted yet replicates efficiently in macrophages and induce neuronal injury. These results demonstrate that HIV-1 variants in the CNS may possess a broader range of biological characteristics than generally appreciated, raise the possibility that X4 strains may participate in AIDS neuropathogenesis, and provide a prototype clade B HIV-1 strain that replicates efficiently in primary macrophages through the exclusive use of CXCR4 as a coreceptor. PMID- 12907389 TI - Decreased neurotropism of nef long terminal repeat (nef/LTR)-deleted simian immunodeficiency virus. AB - Simian immunodeficiency virus (SIV) infection of macaques results in neurological abnormalities similar to those of human immunodeficiency virus (HIV)-associated dementia in humans and is a valuable system for the identification of viral neurotropic and neurovirulence factors. The authors recently established an SIV macaque model where macaques can be infected with wild-type or nef/LTR-deleted SIVmac239 via administration of purified proviral DNA. In this study, the ability of wild-type and nef/LTR-deleted SIV infections to enter the cerebral spinal fluid (CSF) and brain was analyzed. In situ polymerase chain reaction (PCR) readily detected SIV gag DNA-positive cells in the mid-frontal gyrus and basal ganglia of the wild-type SIV-infected macaques, but not in nef/LTR-deleted SIV infected or SIV-uninfected macaques. PCR on extracted DNA confirmed the in situ results, with multiple brain regions of the wild-type SIV-infected macaques positive for both gag and wild-type nef, whereas in the nef/LTR-deleted SIV infected macaques, nef/LTR and gag DNA were undetectable. Further, macaques infected with nef/LTR-deleted SIV, which later became superinfected with wild type SIV, also remained negative for SIV DNA in the brain by both in situ and extracted DNA techniques, despite having high levels of SIV RNA both in the CSF and plasma. This study provides evidence of the inability of nef/LTR-deleted SIV to initiate central nervous system (CNS) infection and suggests that, in the brain regions examined, nef/LTR-deleted viruses have either diminished neurotropism or insufficient systemic viral replication for entry into the CNS. PMID- 12907390 TI - Central nervous system correlates of behavioral deficits following simian immunodeficiency virus infection. AB - Despite the high incidence of cognitive and motor impairment in acquired immunodeficiency syndrome (AIDS) patients, the mechanisms of AIDS-related central nervous system (CNS) pathology are not completely understood. Infection with simian immunodeficiency virus (SIV) in macaques provides an excellent model of AIDS, including human immunodeficiency virus (HIV)-induced CNS pathology and cognitive/behavioral impairment. Co-inoculation with two SIV strains, SIV/17E-Fr and SIV/DeltaB670, accelerates SIV CNS disease, producing SIV encephalitis in over 90% of pig-tailed macaques within 3 months. In the present study, this SIV model was employed to identify cellular and viral correlates of behavioral impairment following SIV infection. Measures of psychomotor speed (simple reaction time), fine motor control (bimanual motor task), and general motor activity (home cage movement) were all adversely affected by SIV disease. Prior to euthanasia, performance was significantly impaired in both a simple reaction time task in 6 of 12 monkeys and a bimanual motor task in 5 of 6 monkeys. All monkeys evaluated (11 of 11) showed significant reductions in spontaneous motor activity. Significant correlations were found between impaired performance on the bimanual motor test and axonal damage (accumulation of beta-amyloid precursor protein in the corpus callosum) as well as increased microglial activation and macrophage infiltration (levels of CD68 and Ham56 immunostaining). These results suggest that axonal damage is related to the behavioral impairment induced by infection with SIV. The axonal damage may result from neuroimmune responses, including microglial and macrophage activation. Therefore, axonal damage may be a morphologic manifestation of neuronal dysfunction that underlies development of behavioral impairment in HIV/SIV CNS infection. PMID- 12907392 TI - The herpes simplex virus (HSV) protein ICP34.5 is a virion component that forms a DNA-binding complex with proliferating cell nuclear antigen and HSV replication proteins. AB - The replicative ability of ICP34.5-null herpes simplex virus (HSV) is cell type and state dependent. In certain cells, ICP34.5 interacts with protein phosphatase 1 to preclude host cell protein synthesis shutoff by dephosphorylation of the eukaryotic initiation factor eIF-2alpha. However, host cell shutoff is not induced by ICP34.5-null HSV in most cells, irrespective of type and state. In general, dividing cells support replication of ICP34.5-null HSV; nondividing cells cannot. Previously the authors showed that ICP34.5 binds to proliferating cell nuclear antigen (PCNA), a protein necessary for cellular DNA replication and repair. Here the authors demonstrate that (1) the interaction between ICP34.5 and PCNA involves two regions of the virus protein; (2) ICP34.5 forms a complex with HSV replication proteins that is DNA binding; (3) at early times in infection, ICP34.5 colocalizes with PCNA and HSV replication proteins in cell nuclei, before accumulating in the cytoplasm; and (4) ICP34.5 is a virion protein. In light of ongoing clinical trials assessing the safety and efficacy of ICP34.5-null HSV, it is vital that the roles of ICP34.5 in HSV replication are understood. The authors propose that in nondividing cells, ICP34.5 is required to switch PCNA from repair to replication mode, a prerequisite for the initiation of HSV replication. PMID- 12907391 TI - Ramified feline microglia selects for distinct variants of feline immunodeficiency virus during early central nervous system infection. AB - It is widely accepted that human immunodeficiency virus (HIV) invades the central nervous system (CNS) shortly after peripheral infection to establish a persistent infection of tissue-resident microglial cells. To what extent this early CNS infection is of pathogenic relevance is a matter of discussion. It is conceivable, however, that infected microglia releases virus variants of enhanced neurotropism and/or neurovirulence compared to peripheral isolates. Moreover, microglial variants may exhibit high resistance to antiviral therapeutics that poorly penetrate into brain tissue. The molecular basis of these biological properties is suspected to be associated with specific sequences in the viral env gene, particularly within the V3 loop. Therefore, we analyzed in the animal model of feline immunodeficiency virus (FIV) infection of cats lentiviral V3 sequences in highly purified microglial cells and blood from acutely infected animals. Compared to the inoculated virus, nucleotide sequence alterations in serum samples were rarely detectable, if at all. In contrast, up to 19 nucleotide exchanges could be identified within FIV V3 from microglia, resulting in a mutation frequency of up to 14.5% with respect to the deduced amino acid sequence. These findings suggest selection of specific virus variants by brain resident target cells that might have implications for antiretroviral drug design. PMID- 12907393 TI - Herpesvirus quiescence in neuronal cells. V: forskolin-responsiveness of the herpes simplex virus type 1 alpha0 promoter and contribution of the putative cAMP response element. AB - The herpes simplex virus (HSV)-1 alpha0 promoter contains a putative cAMP response element (CRE) located at positions -68 to -60 with respect to the initiation of transcription. In this report, the authors examined the functionality of this element using (1) luciferase reporter gene assays in nerve growth factor-differentiated (ND)-PC12 cells and (2) virus-induced activation from quiescently infected (QIF)-PC12 cells. The putative alpha0 CRE was completely eliminated by digestion with the restriction enzyme Tsp45I followed by mung bean nuclease treatment. The mutated region was verified by DNA sequencing and was inserted into the alpha0-luciferase reporter plasmid (pRDalpha0-LUC) creating (pRDalpha0deltaCRE-LUC), and into the HSV-1 genome of strain 17(+)(alpha0deltaCRE). Insertion into both copies of the alpha0 promoter was verified by Southern blot analysis. ND-PC12 cells transfected with pRDalpha0-LUC and pRDalpha0deltaCRE-LUC plasmids responded similarly to forskolin (50 microM), with approximately 250% increases in luciferase activity compared to mock-treated cultures as measured 3 days following treatment. When QIF-PC12 cultures established with HSV-1 strain 17(+) and alpha0deltaCRE were treated with forskolin (50 microM) 17 days post infection, virus was detected in 9/24 (37.5%) and 13/24 (54.2%) of induced cultures by day 8 post treatment, respectively. In contrast, virus was detected in 0/23 and 1/24 (4.2%) of mock-treated cultures by day 8 post treatment for wild-type and mutant viruses, respectively. These findings indicate that the alpha0 promoter is forskolin responsive, the purported CRE of the alpha0 promoter does not confer forskolin responsiveness in ND-PC12 cells, and this element is not required for reactivation of HSV-1 from QIF-PC12 cells. PMID- 12907394 TI - Analysis of the excreted JC virus strains and their potential oral transmission. AB - JC virus (JCV) particles have been detected in urban sewage of divergent geographical areas. In this study, the authors evaluate the genetic characteristics and the infective capabilities of JCV strains in relation to the potential oral transmission of JCV in the population. JCV strains excreted in urine and detected in sewage have been described as presenting archetypal structure of the regulatory region of the viral genome. The regulatory region of JCV viral particles detected in two urban sewage samples have been cloned and characterized. From a total of 40 clones tested, 39 presented archetypal-like regulatory regions, whereas 1 of the clones analyzed presented a tandem repeated structure. Archetypal strains present in the urine of a pregnant woman were able to infect SVG cells, producing infectious virions, as demonstrated by confirmative cell culture, electron microscopy, and in situ DNA hybridization. This is the first description of archetypal JCV productive infection of SVG cells. SVG cells were also successfully infected with Mad-4 JCV viral particles subjected to pH 3 for 1 h at 37 degrees C and to 10 microg/ml of trypsin in the same conditions. A decrease in the viral progeny production was observed when Mad 4 was subjected to acidic pH. Mad-4 did not produce any detectable infection in the enteric cell line CaCo-2. The oral route could represent a significant route of transmission of JCV infections because JCV virions have demonstrated relative resistance in the environment and to some of the conditions present in the gastrointestinal tract. The archetypal strains commonly detected in the environment may be implicated in the transmission of JCV among the population. Sporadic infection with strains presenting tandem repeated structures may have implications in pathogenicity. PMID- 12907396 TI - Putting SARS in perspective: a communication challenge. PMID- 12907395 TI - Simian immunodeficiency virus encephalitis in the white matter and degeneration of the cerebral cortex occur independently in simian immunodeficiency virus infected monkey. AB - Highly active antiretroviral therapy (HAART) has been successful to reduce progression of acquired immunodeficiency syndrome (AIDS). Nevertheless, recent autopsy analysis of the brain from patients with human immunodeficiency virus (HIV)-1 infection reported same or even increasing numbers of AIDS encephalopathy. This insufficient effect of HAART for central nervous system (CNS) complication might be explained by independent pathogenetic processes in lymph node and CNS. We inoculated macaques with three Simian immunodeficiency virus (SIV) strains and investigated relationship between degree of the lymph node pathology and that of AIDS-related brain pathology. Animals infected with T cell-tropic viruses SIVmac239 and SHIV-RT developed typical AIDS pathology in the lymph node 46 to 156 weeks after infection. The cerebral cortex of these animals showed focal or diffuse gliosis, and electron microscopic analysis demonstrated degenerative changes, such as accumulation of dense lamellar bodies in the dendrites and swelling of astrocytic processes. However, there was no evidence of microglial nodules or multinucleated giant cells in the white mater. The animals infected with macrophage-tropic SIV239env/MERT did not develop lymph node pathology of AIDS in the same or longer period of infection. The white mater of the animal, however, showed microglial nodules with multinucleated giant cells, a pathological hallmark of AIDS encephalopathy. SIV immunoreactivity was demonstrated in these giant cells as well as macrophage/microglia cells. On the other hand, there was no abnormality in the cerebral cortex. These findings suggest that there are two independent pathogenetic processes in AIDS encephalopathy: immune response against virus infected macrophage/microglial cells in the white mater without immunodeficiency and cortical degeneration caused in the late stage of AIDS. PMID- 12907398 TI - Media and marijuana: A longitudinal analysis of news media effects on adolescents' marijuana use and related outcomes, 1977-1999. AB - This study examined how aggregate levels of news coverage about marijuana have impacted adolescents' marijuana behavior generally, and through the intervening variables of personal disapproval and perceived harmfulness of marijuana, two variables that existing research has identified as significant predictors of adolescent marijuana use at the aggregate level. It was hypothesized that news coverage of reasons why people should not use marijuana would cause increase in aggregate marijuana abstinence, perceived harmfulness, and personal disapproval. Conversely, news coverage of positive aspects of marijuana use would cause decreases in marijuana abstinence, perceived harmfulness, and personal disapproval. Results of distributed lagged time-series regression and non-linear modeling offered support for two of the three proposed hypotheses. Aggregate media coverage explained a significant portion of the variation in adolescents' abstinence from marijuana use over time. It also explained a significant portion of the variation in personal disapproval of marijuana. Personal disapproval was found to partially mediate the relationship between media coverage and marijuana abstinence. Implications for the conceptualization of media effects on health behaviors are discussed. PMID- 12907399 TI - Health related content in prime-time television programming. AB - This study identified, content analyzed, and described the health-related content (HRC) presented in the top-ranked prime-time network programs for the age 2 to 11 year-old category. The results reveal that viewers see one HRC-containing scene approximately every four minutes. Foods and alcoholic beverages are frequently shown and consumed. Smoking, drug abuse, and promiscuous sex are relatively uncommon. In most scenes, the HRC is not directly related to the main point of the scene. The characters involved in HRC-containing scenes tend to be slender white men. In addition, many of the HRC-containing scenes portray negative health behaviors. PMID- 12907400 TI - Stigmatizing smokers: public sentiment toward cigarette smoking and its relationship to smoking behaviors. AB - Public sentiment in the United States has been evolving against cigarette smoking. Providing support for stronger tobacco control legislation, unfavorable public sentiment has contributed to the decrease in the size of the smoking population in this country. The present study hypothesizes that the unfavorable public sentiment may also discourage cigarette smoking by creating an unfavorable "smoking climate" in which smoking is socially rejected as a deviant behavior. Analyses of several secondary data-sets provided evidence that smoking rates are lower in the states where the public holds relatively unfavorable sentiment toward cigarette smoking. The relationship between public sentiment and smoking rates was significant even after controlling for the effects of state-level tobacco control measures, such as cigarette taxes and smoking restrictions in private workplaces and restaurants. We also found that smokers who have experienced unfavorable public sentiment are more willing to quit smoking than those who have not, supporting the hypothesized effects of antismoking public sentiment on smoking behaviors. PMID- 12907401 TI - Stroke knowledge and barriers to stroke prevention among African Americans: implications for health communication. AB - Stroke is a major cause of death and disability, especially among African Americans. Yet research on stroke knowledge and barriers to stroke prevention among African Americans is limited. This study used a 50-item questionnaire to conduct structured telephone interviews with 379 African American adults, 50 years or older. The questionnaire included questions on stroke knowledge, stroke risk behaviors, and barriers to stroke prevention. A stroke knowledge score was computed by assigning points for correct responses on knowledge items. The average stroke knowledge score of participants was 10.9, out of a maximum possible score of 27. Stroke knowledge was significantly related to the presence of hypertension, heart disease, diabetes, and family history of stroke. College education was significantly associated with older respondents' stroke knowledge. Younger college-educated respondents had more knowledge about the risky behaviors that lead to stroke than those with less education. Respondents preferred hospitals to churches or senior centers for receiving stroke information and for learning about stroke from their physicians. Stress and poor financial status were most frequently reported as barriers to stroke prevention. These findings have implications for developing more effective strategies for educating African Americans about stroke prevention. PMID- 12907402 TI - Order from chaos: organizational aspects of information, education, and communication (a case study from Mali). PMID- 12907403 TI - Nutritional enrichment of fresh apple (Royal Gala) by vacuum impregnation. AB - This study evaluates the use of vacuum impregnation (VI) for developing nutritionally fortified fresh cut apples (Royal Gala). Cut apples were immersed in diluted high fructose corn syrup (20% w/w or 50% w/w) containing calcium or zinc. A vacuum pressure of 50 mmHg was applied for 15 min following atmospheric pressure restoration for 30 min while samples remained in the VI solution. Nutraceutical content and physicochemical properties of the apples immediately after VI were determined. Storability of VI apples at 6 degrees C and 90% relative humidity was also studied based on the color and texture of apples. Results indicated that 15-20% of the Daily Reference Intake of calcium and above 40% of the Daily Reference Intake of zinc could be obtained in 200 g fresh cut apples. VI treatments in 20% w/w high fructose corn syrup solutions had little effects on the physicochemical properties of apples. Storage study showed that VI with zinc significantly improved color stability, and calcium enhanced the firmness of the apples. PMID- 12907404 TI - Small fish is an important dietary source of vitamin A and calcium in rural Bangladesh. AB - Fish plays an important role in the Bangladeshi diet, constituting the main animal food in rural households. Fish consumption is dominated by wild small (length<25 cm) indigenous fish species (SIS). A study was conducted to evaluate the dietary contribution of vitamin A and calcium from fish in rural Bangladesh. Seasonal fish consumption was surveyed in 84 rural households in Kishoreganj district using a 5 day recall method. Fifty-nine of the households practised aquaculture, producing carps and SIS in small ponds. The intra-household distribution of a standardised fish dish was surveyed in 20 households by weighing. The contribution of vitamin A and calcium from fish was calculated as a nutrient contribution ratio, expressing the nutrient intake from fish relative to the recommended intake at household level. The mean amount of fish consumed in all households was 37 g raw edible parts/person/day (median=27) in July 1997. 82 g raw edible parts/person/day (median=64) in October 1997, and 55 g raw edible parts/person/day (median=42) in February 1998. A total of 44 fish species was recorded in the diet. SIS contributed 84% of the total fish intake. Fish bought in the local markets was the most important source of fish in both fish-producing and non-fish-producing households. The intra-household fish distribution showed that the heads of the households (all males) were favoured over females and other males. In October, the; mean nutrient contribution ratio for vitamin A was 40% (median=23%) and for calcium was 32% (median=26%). SIS contributed more than 99% of vitamin A and calcium intakes from the total fish intake. Mola, a vitamin A rich SIS, was produced in 34 of the fish-producing households. Mola harvested from the ponds and consumed in the households contributed, on average, 21% (median=18%) of the household recommended intake of vitamin A in a 7 month period. SIS is an important and perhaps irreplaceable dietary source of vitamin A and calcium, and promotion of the availability and accessibility of SIS for the population in rural Bangladesh should be given priority. PMID- 12907405 TI - Development of a nutritious low viscosity weaning mix using natural ingredients and microbial amylases. AB - A nutritious weaning food was developed using natural ingredients; namely, staple cereals and pulses, groundnut, Spirulina and gooseberry. The nutritive value of the developed weaning mix was found to be better than a commercial mix. The viscosity of the mixes was reduced by the addition of bacterial and fungal amylases. Addition of amylases at a concentration of 0.1-0.4% drastically reduced the viscosity in all the formulated mixes. The maximum viscosity reduction effect was evident at 0.2% for bacterial amylase and at 0.4% for fungal amylases. PMID- 12907406 TI - Antioxidant activity of the anthocyanin from carrot (Daucus carota) callus culture. AB - Antioxidant properties of water-soluble anthocyanin obtained from the carrot (Daucus carota) callus cultures were evaluated by the estimation of the amount of hydroperoxide formed by the autoxidation of linolenic acid at pH 2, 4 and 7. A major anthocyanin, cyanidin-3-lathyroside, was isolated and the antioxidant property was compared with malvidin, peonidin delphinidin and alpha-tocopherol. The malvidin, peonidin and delphinidin were obtained from grapes, Chinese mustard and egg-plant. At pH 2 and 4 the anthocyanin was better than malvidin and peonidin. The results indicate that anthocyanin obtained from carrot showed an antioxidation activity stronger than malvidin, peonidin and alpha-tocopherol, and weaker than delpinidin at pH 2, 4 and 7 by the linolenic acid auto-oxidation system. PMID- 12907407 TI - Organic food: nutritious food or food for thought? A review of the evidence. AB - Apparently, one of the primary reasons for purchasing organic food is the perception that it is more nutritious than conventional food. Given the increasing interest towards organic food products, it is imperative to review the existing literature concerning the nutritional value of the produce, and to determine to what extent are consumer expectations met. There are only few well controlled studies that are capable of making a valid comparison and, therefore, compilation of the results is difficult and generalisation of the conclusions should be made with caution. In spite of these limitations, however, some differences can be identified. Although there is little evidence that organic and conventional foods differ in respect to the concentrations of the various micronutrients (vitamins, minerals and trace elements), there seems to be a slight trend towards higher ascorbic acid content in organically grown leafy vegetables and potatoes. There is also a trend towards lower protein concentration but of higher quality in some organic vegetables and cereal crops. With respect to the rest of the nutrients and the other food groups, existing evidence is inadequate to allow for valid conclusions. Finally, animal feeding experiments indicate that animal health and reproductive performance are slightly improved when they are organically fed. A similar finding has not yet been identified in humans. Several important directions can be highlighted for future research; it seems, however, that despite any differences, a well-balanced diet can equally improve health regardless of its organic or conventional origin. PMID- 12907408 TI - Effect of cooking on oxalate content of pulses using an enzymatic procedure. AB - An enzymatic method proposed for the determination of oxalate in urine is adapted for the estimation of soluble oxalate content in beans, chickpeas and lentils. Oxalates were extracted in water by refluxing for 2 h. The method is based on the oxidation of oxalate by the oxidase and the determination of the resulting hydrogen peroxide, which in presence of peroxidase, 3-methyl-2 benzotiazinolone and 3-dimethylamino benzoic, gives an indamine compound with an absorption maximum at 590 nm. The linearity (from 0.015 to 0.6 mM) of the method is adequate to the analysis of oxalate contents in pulses, and the inter-day precision of the method expressed as relative standard deviation was good (3.01%), with an accuracy of (98.7+/-2.5%) estimated by recovery assays. This method is applied to estimate the losses of soluble oxalate as a consequence of cooking. All the cooking procedures reduce the soluble oxalate content, but microwave and industrial procedures are more effective than traditional domestic cooking. PMID- 12907409 TI - Docosahexaenoic acid level of the breast milk of some Filipino women. AB - This study was conducted to determine the docosahexaenoic acid (DHA) level of the breast milk of 100 Filipino women as affected by diet. The subject distribution was patterned after the 1997 Family and Income Expenditure Survey of the National Statistics Office regarding the total number of families, and the total and average family income and expenditures by income class in an urban area. The subjects were asked to complete a 3-day food record and food frequency questionnaire to ascertain the nutrient content of their food intake and approximate eating habits. Hind milk was drawn manually by means of a fabricated glass breast pump and collected in polypropylene vials. The milk samples were stored in a freezer maintained at -25+/-2 degrees C until they were transported to the University of Montreal for fatty acid composition. The milk was methylated using the Lepage and Roy method. The obtained fatty acid methyl esters were analyzed by gas chromatography. Results showed that the milk samples contain an average of 188.34 microg DHA/ml milk, while %DHA of the samples is 0.65%. Regression analysis revealed that mean protein intake of the subjects was a determinant of the DHA level in the milk samples. PMID- 12907410 TI - The effect of a micronutrient-fortified complementary food on micronutrient status, growth and development of 6- to 12-month-old disadvantaged urban South African infants. AB - The study was conducted to look at the effectiveness of a multimicronutrient fortified complementary food on the micronutrient status, linear growth and psychomotor development of 6- to 12-month-old infants from a black urban disadvantaged community in the Western Cape, South Africa. The study was designed as an intervention study. In both the experimental and control groups, serum retinol concentration showed a decline over the intervention period of 6 months. The decline was less pronounced in the experimental group. This resulted in a significantly (P<005) higher serum retinol concentration at 12 months in the experimental group (26.8+/-5.8 microg/dl) compared with the control group (21.4+/ 5 microg/dl). Serum iron concentration also declined over the intervention period. The decline was less pronounced in the experimental group. No difference was observed in haemoglobin levels between the groups at 12 months. Serum zinc concentration did not differ significantly between the two groups at follow up. Weight gain over the 6 months period did not differ significantly between the experimental (2.1+/-0.9 kg) and control groups (2.1+/-1.2 kg). There was no difference in linear growth between the experimental (10.0+/-1.5 cm) and control group (10.1+/-2.1 cm) at the end of the follow-up period. Weight and length at 6 months significantly predicted weight and length at 12 months. No difference was observed in psychomotor developmental scores between the two groups after 6 months of intervention. Introducing a multimicronutrient-fortified complementary food into the diet of 6- to 12-month-old infants seemed to have an arresting effect on declining serum retinol and iron concentration in the experimental group. No benefit was observed in serum zinc concentration, linear growth and psychomotor development. PMID- 12907412 TI - Mitochondrial proliferation in the permanent vs. temporary cold: enzyme activities and mRNA levels in Antarctic and temperate zoarcid fish. AB - Adjustments in mitochondrial properties and capacities are crucial in acclimatization to seasonal cold and in evolutionary cold adaptation of marine ectotherms. Although long-term compensatory increments in aerobic capacity of fish tissues have frequently been described in response to cold, much less is known about transitional phases and gene expression patterns involved. We investigated the time course of adjustment to acute cold in liver of eurythermal eelpout Zoarces viviparus. Whereas citrate synthase (CS) activity rose progressively in liver, cytochrome c oxidase (COX) activity was not altered during cold acclimation. Species-specific RNA probes were used to determine mRNA levels. CS mRNA (nuclear encoded) displayed a delayed, transient increase in response to cold, such that transcript levels did not parallel the change in enzyme activity. The enzyme activities and mRNA levels in the confamilial Antarctic Pachycara brachycephalum indicate cold compensation of CS activity in this cold-adapted species. The ratio of CS and COX activities was elevated in acclimation and adaptation to cold, indicating enhanced citrate synthesis over respiratory chain capacities in cold-adapted liver mitochondria. This may support enhanced lipid synthesis typically found in cold. The ratio of enzyme activity and transcript levels differed largely between Z. viviparus populations from the Baltic and North Seas, indicating the influence of unidentified parameters other than temperature. Transcript levels may not be tightly correlated with enzyme activities during thermal adaptation and thereafter. The time course of the acclimation process indicates that regulation at the translational and posttranslational levels predominates in adjustment to moderate thermal challenges. PMID- 12907413 TI - Effects of central oxytocin receptor blockade on water and saline intake, mean arterial pressure, and c-Fos expression in rats. AB - Central injection of ANG II has been proposed to have dual effects on salt appetite including a direct stimulatory effect and an indirect inhibitory effect through an activation of central oxytocinergic neurons. The inhibition was demonstrated by pretreating rats with central ornithine vasotocin (OVT; oxytocin antagonist) 30 min before a central ANG II injection. The OVT pretreatment produced a large increase in ANG II-induced saline intake. The present paper reports a failure to replicate that influential experiment. However, we also report for the first time that OVT by itself: 1) provokes drinking of both water and saline solution with a latency almost as short as that produced by ANG II; 2) produces a mild pressor response; and 3) increases c-Fos expression in the organum vasculosum laminae terminalis (OVLT) and the median preoptic nucleus (MnPO). Oxytocin activity may provide an inhibitory control of drinking responses as has been suggested, but the inhibition is tonic and includes both water and saline drinking. Inhibition of this tonic activity may stimulate drinking by increasing neural activity in the OVLT and MnPO. PMID- 12907414 TI - Effects of the opioid antagonist naltrexone on feeding induced by DAMGO in the ventral tegmental area and in the nucleus accumbens shell region in the rat. AB - The nucleus accumbens shell region (sNAcc) and the ventral tegmental area (VTA) are two major nodes in the mesolimbic dopamine pathway, which mediates reward for various survival behaviors, including feeding. Opioids increase and maintain food intake when injected peripherally and centrally. Opioids in the VTA cause increased release of dopamine in the sNAcc, and when injected into either site, cause an increase in food intake. Animals in this study were double cannulated in the VTA and in the sNAcc and injected with various combinations of naltrexone (NTX) (2.5, 5, and 25 microg/side) and Tyr-d-Ala-Gly-(Me)Phe-Gly-ol (DAMGO) (0.1, 0.3, 1, 3, and 5 nmol/side) in both sites. DAMGO was found to dose dependently increase intake to an equal extent when injected into either site. DAMGO-induced increases in food intake when injected into the VTA were blocked to control levels with the highest dose of NTX injected bilaterally into the sNAcc; however, increases in intake when injected into the sNAcc were blocked only partially by the highest dose of NTX injected bilaterally into the VTA. These results indicate opioid-opioid communication between the two sites; however, the communication may be quite indirect, requiring other sites and transmitters to elicit a change in behavior. PMID- 12907415 TI - Cortisol alters carbonic anhydrase-mediated renal sulfate secretion. AB - Active transepithelial sulfate secretion rate by winter flounder renal proximal tubule epithelium in primary culture (fPTC) is dependent on intracellular carbonic anhydrase (CA) and enhanced by cortisol. To further evaluate this relationship, a partial cDNA clone (327 bp) of carbonic anhydrase II (CAII) with high sequence similarity to CAII from numerous species including fish, chicken, and human was obtained from fPTCs. The majority of CA activity and CAII protein was present in the cytosol of fPTCs; however, significant amounts of both (in addition to SDS-resistant CA activity, i.e., CAIV-like isoform) were present in concentrated plasma membranes. CAII from concentrated membranes migrated differently than purified CAII on nondenaturing PAGE gels, suggesting that CAII associates with another membrane component. Treatment of fPTCs with the cell soluble CA inhibitor methazolamide (100 microM) caused a 58% reduction in active transepithelial SO4(2-) secretion. fPTCs that were previously cultured under high cortisol concentrations, when subjected to 5 days of low physiological levels of cortisol, had decreased CA activity (28%), CAII protein abundance (65%), and net active SO4(2-) secretion (28%), with no effect on epithelial differentiation. Methazolamide and low-cortisol treatment in combination inhibited net active SO4(2-) secretion 56%, which was not different than the effect of methazolamide treatment alone. These data indicate that cortisol directly increases renal CA activity, CAII protein abundance, and CA-dependent SO4(2-) secretion in the marine teleost renal proximal tubule. PMID- 12907416 TI - Directionality of coupling of physiological subsystems: age-related changes of cardiorespiratory interaction during different sleep stages in babies. AB - Activity of many physiological subsystems has a well-expressed rhythmic character. Often, a dependency between physiological rhythms is established due to interaction between the corresponding subsystems. Traditional methods of data analysis allow one to quantify the strength of interaction but not the causal interrelation that is indispensable for understanding the mechanisms of interaction. Here we present a recently developed method for quantification of coupling direction and apply it to an important problem. Namely, we study the mutual influence of respiratory and cardiovascular rhythms in healthy newborns within the first 6 mo of life in quiet and active sleep. We find an age-related change of the coupling direction: the interaction is nearly symmetric during the first days and becomes practically unidirectional (from respiration to heart rhythm) at the age of 6 mo. Next, we show that the direction of interaction is mainly determined by respiratory frequency. If the latter is less than approximately 0.6 Hz, the interaction occurs dominantly from respiration to heart. With higher respiratory frequencies that only occur at very young ages, the dominating direction is less pronounced or even abolished. The observed dependencies are not related to sleep stage, suggesting that the coupling direction is determined by system-inherent dynamical processes, rather than by functional modulations. The directional analysis may be applied to other interacting narrow band oscillatory systems, e.g., in the central nervous system. Thus it is an important step forward in revealing and understanding causal mechanisms of interactions. PMID- 12907417 TI - Balanced expression of mitochondrial apoptosis regulatory proteins correlates with long-term survival of cardiac allografts. AB - Abnormal regulation of apoptosis is observed in ischemic injury and may contribute to the pathogenesis of atherosclerosis. However, its role in cardiac allograft vasculopathy (CAV), the fundamental lesion of chronic rejection (CR) in heart transplantation, remains uncertain. To clarify this issue, apoptosis was quantitated in myocardium and coronary arteries from 5 cardiac allograft donors (NL) and explanted hearts of 24 patients with ischemic cardiomyopathy (IsCM) and 15 patients with CR. Tissue samples were analyzed via end-labeling fragmented DNA [via deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL)] and immunoblotting for activated caspase-3 and -9. Myocyte apoptosis assessed by TUNEL was similarly increased over NL (0.21%) in both the CR (0.88%; P < 0.01) and IsCM (0.88%; P < 0.01) groups. Activated caspase-9 levels were significantly higher in CR (14.7%) compared with IsCM (6.9%; P < 0.01) and NL (0%) groups, whereas activated caspase-3 levels were similarly elevated in both CR and IsCM (7.8 and 6.5% vs. 0% in NL; P < 0.01 and P < 0.05) groups. Expression of myocardial Bcl-2 and Bax was increased in CR compared with both NL (Bax, 4.3 fold; P < 0.01; Bcl-2, 5.9-fold; P < 0.01) and IsCM (IsCM: Bax, 2.2-fold; P < 0.05; Bcl-2, 3.2-fold; P < 0.01) groups. The rate of apoptosis and the Bcl-2/Bax ratio independently correlated to graft survival in CR (activation of caspase-9: r = 0.87; P < 0.01; Bcl-2/Bax: r = 0.57; P = 0.05). Compared with native atherosclerosis, coronary arteries with CAV showed more medial apoptosis (7.8 fold; P < 0.01) and higher Bcl-2 levels (5.1-fold; P < 0.01) with lower Bax levels (threefold; P < 0.05) in the intima. These results indicate that abnormal Bcl-2 and Bax expression in myocardium and coronary arteries of cardiac allografts with CR is distinct from that in IsCM and suggest that balancing Bcl-2 to Bax in transplanted hearts promotes long-term graft survival. PMID- 12907418 TI - Localization of the permeability barrier to solutes in isolated arteries by confocal microscopy. AB - Endothelial cells are covered by a surface layer of membrane-associated proteoglycans, glycosaminoglycans, glycoproteins, glycolipids, and associated plasma proteins. This layer may limit transendothelial solute transport. We determined dimension and transport properties of this endothelial surface layer (ESL) in isolated arteries. Rat mesenteric small arteries (diameter approximately 150 microm) were isolated and cannulated with a double-barreled -pipette on the inlet side and a regular pipette on the outlet side. Dynamics and localization of intra-arterial fluorescence by FITC-labeled dextrans (FITC-Deltas) and the endothelial membrane dye DiI were determined with confocal microscopy. Large FITC Delta (148 kDa) filled a core volume inside the arteries within 1 min but was excluded from a 2.6 +/- 0.5-microm-wide region on the luminal side of the endothelium during 30 min of dye perfusion. Medium FITC-Delta (50.7 kDa) slowly penetrated this ESL within 30 min but did not permeate into the arterial wall. Small FITC-Delta (4.4 kDa) quickly passed the ESL and accumulated in the arterial wall. Prolonged luminal fluorochrome illumination with a bright mercury lamp destroyed the approximately 3-microm exclusion zone for FITC-Delta 148 within a few minutes. This study demonstrates the presence of a thick ESL that contributes to the permeability barrier to solutes. The layer is sensitive to phototoxic stress, and its damage could form an early event in atherosclerosis. PMID- 12907419 TI - SLControl: PC-based data acquisition and analysis for muscle mechanics. AB - SLControl is a computerized data acquisition and analysis system that was developed in our laboratory to help perform mechanical experiments using striated muscle preparations. It consists of a computer program (Windows 2000 or later) and a commercially available data acquisition board (16-bit resolution, DAP5216a, Microstar Laboratories, Bellevue, WA). Signals from the user's existing equipment representing force, fiber length (FL), and (if desired) sarcomere length (SL) are connected to the system through standard Bayonet Neill Concelman cables and saved to data files for later analysis. Output signals from the board control FL and trigger additional equipment, e.g., flash lamps. Windows dialogs drive several different experimental protocols, including slack tests and rate of tension recovery measurements. Precise measurements of muscle stiffness and force velocity/power characteristics can also be accomplished using SL and tension control, respectively. In these situations, the FL command signal is updated in real time (at rates > or =2.5 kHz) in response to changes in the measured SL or force signals. Data files can be exported as raw text or analyzed within SLControl with the use of built-in tools for cursor analysis, digital filtering, curve fitting, etc. The software is available for free download at http://www.slcontrol.com. PMID- 12907420 TI - Spatial heterogeneity of action potential alternans during global ischemia in the rabbit heart. AB - Cardiac ischemia causes beat-to-beat fluctuation in action potential duration (APD) alternans, which leads to T wave alternans and arrhythmias. Occurrence of APD alternans that is out of phase at two sites is especially important, but most APD alternans studies have involved rapid pacing of normal myocardium rather than ischemia. To determine the spatial features of APD alternans during ischemia, blood-perfused rabbit hearts were stained with 4-[beta-[2(di-n-butylamino)-6 napthyl]vinyl]pyridinium (di-4-ANEPPS) and imaged with a high-resolution camera. Hearts were perfused with oxygenated Tyrode solution at 37 degrees C for staining and then switched to a 50:50% blood/Tyrode mixture. Hearts were paced from the right ventricle at 3/s, and made ischemic by stopping flow for 6 min. Images of 10,000 pixels were obtained at 300 frames/s. Motion artifact was controlled by immobilization and by manual selection of undistorted single-pixel records. Upstroke propagation and conduction isochrones were displayed by computerized image processing. APD alternans was demonstrated in six of seven hearts, and was out of phase in different regions of the image in three hearts. The largest spatial variation in the onset of depolarization to 50% repolarization (APD50) was 155%. This caused beat-to-beat reversal of repolarization. An alternans map could be constructed for well-immobilized portions of the image. There were discrete regions of APD alternans separated by a boundary, as occurs with intracellular Ca2+ concentration alternans. Pixels as close together as 1.1 mm showed an APD alternans that was out of phase. The out-of-phase APD alternans was not due to conduction alternans, as shown by upstroke intervals and conduction isochrones. This contrasts with rapid pacing, where a causal relationship appears to exist. These new observations suggest distinct mechanisms for the genesis of arrhythmias during ischemia. PMID- 12907421 TI - Does whole body autoregulation mediate the hemodynamic responses to increased dietary salt in rats with clamped ANG II? AB - The present study was conducted to test the hypothesis that salt-dependent hypertension, in rats with an unresponsive renin-angiotensin system, is characterized by a "whole body autoregulation" hemodynamic profile. To test this hypothesis, rats were chronically instrumented to continuously measure cardiac output (CO) and arterial pressure (AP). A venous catheter was implanted for infusion of saline vehicle (Veh; n = 8) or treatment [enalapril (2 mg.kg-1.day-1) plus ANG II: ANG-NORM (5 ng.kg-1.min-1 ANG II, n = 8) or ANG-HI (10 ng.kg-1.min-1 ANG II, n = 9)] to pharmacologically clamp plasma ANG II. After a 10-day recovery period on a 0.1% NaCl diet, AP and CO were measured continuously for 5 days of control (0.1% NaCl), 7 days of high salt (4.0% NaCl), and 5 days of recovery (0.1% NaCl). Hemodynamics did not change in the Veh group at any time. AP increased by approximately 20 mmHg in the ANG-NORM and ANG-HI groups when NaCl was increased. Hypertension was mediated by an increase in CO of approximately 12% at steady state, with no change in total peripheral resistance (TPR) during the high salt period. AP returned to control levels when dietary sodium was decreased, mediated by a approximately 10% decrease in TPR, with CO remaining elevated. There was no difference in the hemodynamic responses to increased salt between the ANG-HI and ANG-NORM groups. We conclude that the whole body autoregulation hypothesis does not explain the hemodynamic profile of salt dependent hypertension in rats with an unresponsive renin-angiotensin system. PMID- 12907423 TI - Angiotensin II attenuates functional hyperemia in the mouse somatosensory cortex. AB - We investigated whether angiotensin II (ANG II), a peptide that plays a central role in the genesis of hypertension, alters the coupling between synaptic activity and cerebral blood flow (CBF), a critical homeostatic mechanism that assures adequate cerebral perfusion to active brain regions. The somatosensory cortex was activated by stroking the facial whiskers in anesthetized C57BL/6J mice while local CBF was recorded by laser-Doppler flowmetry. Intravenous ANG II infusion (0.25 mug.kg-1.min-1) increased mean arterial pressure (MAP) from 82 +/- 2 to 102 +/- 3 mmHg (P < 0.05) without affecting resting CBF (P > 0.05). ANG II attenuated the CBF increase produced by whisker stimulation by 65% (P < 0.05) but did not affect the response to hypercapnia or to neocortical application of the nitric oxide donor S-nitroso-N-acetyl penicillamine (P > 0.05). The effect of ANG II on functional hyperemia persisted if the elevation in MAP was offset by controlled hemorrhage or prevented by topical application of the peptide to the activated cortex. ANG II did not reduce the amplitude of the P1 wave of the field potentials evoked by whisker stimulation (P > 0.05). Infusion of phenylephrine increased MAP (P > 0.05 from ANG II) but did not alter the functional hyperemic response (P > 0.05). The data suggest that ANG II alters the coupling between CBF and neural activity. The mechanisms of the effect are not related to the elevation in MAP and/or to inhibition of the synaptic activity evoked by whisker stimulation. The imbalance between CBF and neural activity induced by ANG II may alter the homeostasis of the neuronal microenvironment and contribute to brain dysfunction during ANG II-induced hypertension. PMID- 12907422 TI - 11,12,15-Trihydroxyeicosatrienoic acid mediates ACh-induced relaxations in rabbit aorta. AB - Rabbit aortic endothelium metabolizes arachidonic acid (AA) by the 15 lipoxygenase pathway to vasodilatory eicosanoids, hydroxyepoxyeicosatrienoic acids (HEETAs), and trihydroxyeicosatrienoic acids (THETAs). The present study determined the chemical identity of the vasoactive THETA and investigated its role in ACh-induced relaxation in the rabbit aorta. AA caused endothelium dependent, concentration-related relaxations of the rabbit aorta. Increasing the extracellular KCl concentration from 4.8 to 20 mM inhibited the relaxations to AA by approximately 60%, thereby implicating K+-channel activation in the relaxations. In addition, AA caused an endothelium-dependent hyperpolarization of aortic smooth muscle from -39.6 +/- 2.7 to -56.1 +/- 3.4 mV. In rabbit aortic rings, [14C]AA was metabolized to prostaglandins, HEETAs, THETAs, and 15 hydroxyeicosatetraenoic acid. Additional purification of the THETAs by HPLC resolved the mixture into its 14C-labeled products. Gas chromatography/mass spectrometry identified the metabolites as isomers of 11,12,15-THETA and 11,14,15 THETA. The 11,12,15-THETA relaxed and hyperpolarized the rabbit aorta, whereas 11,14,15-THETA had no vasoactive effect. The relaxations to 11,12,15-THETA were blocked by 20 mM KCl. In aortic rings pretreated with inhibitors of nitric oxide and prostaglandin synthesis, ACh caused a concentration-related relaxation that was completely blocked by 20 mM KCl. Pretreatment with the phospholipase A2 inhibitors mepacrine and 7,7-dimethyl-5,8-eicosadienoic acid, the lipoxygenase inhibitors cinnamyl-3,4-dihydroxy-alpha-cyanocinnamate, nordihydroguaiaretic acid, and ebselen, or the hydroperoxide isomerase inhibitors miconazole and clotrimazole also blocked ACh-induced relaxations. ACh caused a threefold increase in THETA release. These studies indicate that AA is metabolized by endothelial cells to 11,12,15-THETA, which activates K+ channels to hyperpolarize the aortic smooth muscle membrane and induce relaxation. Additionally, this lipoxygenase pathway mediates the nonnitric oxide, nonprostaglandin relaxations to ACh in the rabbit aorta by acting as a source of an endothelium-derived hyperpolarizing factor. PMID- 12907424 TI - Differential vasoconstrictions induced by angiotensin II: role of AT1 and AT2 receptors in isolated C57BL/6J mouse blood vessels. AB - Genetically altered mice are increasingly used as experimental models. However, ANG II responses in mouse blood vessels have not been well defined. Therefore, the aim of this study was to determine the role of ANG II in regulating major blood vessels in C57/BL6J mice with isometric force measurements. Our results showed that in mouse abdominal aorta ANG II induced a concentration-dependent contraction (EC50 4.6 nM) with a maximum contraction of 75.1 +/- 4.9% at 100 nM compared with that of 60 mM K+. Similarly, femoral artery also exhibited a contractile response of 76.0 +/- 3.4% to the maximum concentration of ANG II (100 nM). In contrast, ANG II (100 nM)-induced contraction was significantly less in carotid artery (24.5 +/- 6.6%) and only minimal (3.5 +/- 0.31%) in thoracic aorta. The nitric oxide synthase inhibitor N omega-nitro-L-arginine methyl ester and the AT2 antagonist PD-123319 failed to enhance ANG II-induced contractions. However, an AT1 antagonist, losartan (10 microM), completely inhibited ANG II (100 nM) response in abdominal aorta and carotid artery. An AT1 agonist, [Sar1] ANG II (100 nM), behaved similarly to ANG II (100 nM) in abdominal aorta and carotid artery. RT-PCR analyses showed that mouse thoracic aorta has a significantly lower AT1 mRNA level than abdominal aorta. These results demonstrate that major mouse vessels exhibit differential contractions to ANG II, possibly because of varied AT1 receptor levels. PMID- 12907425 TI - A selective inducible NOS dimerization inhibitor prevents systemic, cardiac, and pulmonary hemodynamic dysfunction in endotoxemic mice. AB - Increased nitric oxide (NO) production by inducible NO synthase (NOS2), an obligate homodimer, is implicated in the cardiovascular sequelae of sepsis. We tested the ability of a highly selective NOS2 dimerization inhibitor (BBS-2) to prevent endotoxin-induced systemic hypotension, myocardial dysfunction, and impaired hypoxic pulmonary vasoconstriction (HPV) in mice. Mice were challenged with Escherichia coli endotoxin before treatment with BBS-2 or vehicle. Systemic blood pressure was measured before and 4 and 7 h after endotoxin challenge, and echocardiographic parameters of myocardial function were measured before and 7 h after endotoxin challenge. The pulmonary vasoconstrictor response to left mainstem bronchus occlusion, which is a measure of HPV, was studied 22 h after endotoxin challenge. BBS-2 treatment alone did not alter baseline hemodynamics. BBS-2 treatment blocked NOS2 dimerization and completely inhibited the endotoxin induced increase of plasma nitrate and nitrite levels. Treatment with BBS-2 after endotoxin administration prevented systemic hypotension and attenuated myocardial dysfunction. BBS-2 also prevented endotoxin-induced impairment of HPV. In contrast, treatment with NG-nitro-l-arginine methyl ester, which is an inhibitor of all three NOS isoforms, prevented the systemic hypotension but further aggravated the myocardial dysfunction associated with endotoxin challenge. Treatment with BBS-2 prevented endotoxin from causing key features of cardiovascular dysfunction in endotoxemic mice. Selective inhibition of NOS2 dimerization with BBS-2, while sparing the activities of other NOS isoforms, may prove to be a useful treatment strategy in sepsis. PMID- 12907426 TI - Role of interleukin 12 in hypercholesterolemia-induced inflammation. AB - We have previously shown that T lymphocytes and interferon-gamma are involved in hypercholesterolemia-induced leukocyte adhesion to vascular endothelium. This study assessed the contribution of interleukin 12 (IL-12) to these hypercholesterolemia-induced inflammatory responses. Intravital videomicroscopy was used to quantify leukocyte adhesion and emigration and oxidant stress (dihydrorhodamine oxidation) in unstimulated cremasteric venules (wall shear rate > or =500 s-1) of wild-type (WT) C57Bl/6, lymphocyte-deficient [recombinase activating gene knockout (RAG1-/-)], and IL-12-deficient (p35-/- and p40-/-; p35 and p40 are the two subunits of active IL-12) mice on either a normal (ND) or high-cholesterol (HC) diet for 2 wk. RAG1-/--HC mice received splenocytes from WT HC (WT --> RAG1-/-), p35-/--HC (p35-/- --> RAG1-/-), or p40-/--HC (p40-/- --> RAG1-/-) mice. Compared with WT-ND mice, WT-HC mice exhibited exaggerated leukocyte adherence and emigration as well as increased dihydrorhodamine oxidation. The enhanced leukocyte recruitment was absent in the RAG1-/--ND, p35-/ -ND, and p40-/--ND groups. Hypercholesterolemia-induced leukocyte adherence and emigration were attenuated in RAG1-/--HC vs. WT-HC mice but were similar to ND mice. Furthermore, compared with WT-HC animals, p35-/--HC and p40-/--HC mice showed significantly lower leukocyte adhesion and tissue oxidant stress responses, but these values were comparable to ND mice. Leukocyte adherence and emigration in WT --> RAG1-/- mice were similar to responses of WT-HC mice. However, p35-/- --> RAG1-/- mice had lower levels of adherence and emigration vs. the WT --> RAG1-/- and WT-HC groups. Elevated levels of leukocyte adherence and emigration were restored by approximately 50% toward WT-HC levels in p40-/- --> RAG1-/- mice. These findings implicate IL-12 in the inflammatory responses observed in the venules of hypercholesterolemic mice. PMID- 12907428 TI - Static filling pressure in patients during induced ventricular fibrillation. AB - The static pressure resulting after the cessation of flow is thought to reflect the filling of the cardiovascular system. In the past, static filling pressures or mean circulatory filling pressures have only been reported in experimental animals and in human corpses, respectively. We investigated arterial and central venous pressures in supine, anesthetized humans with longer fibrillation/defibrillation sequences (FDSs) during cardioverter/defibrillator implantation. In 82 patients, the average number of FDSs was 4 +/- 2 (mean +/- SD), and their duration was 13 +/- 2 s. In a total of 323 FDSs, arterial blood pressure decreased with a time constant of 2.9 +/- 1.0 s from 77.5 +/- 34.4 to 24.2 +/- 5.3 mmHg. Central venous pressure increased with a time constant of 3.6 +/- 1.3 s from 7.5 +/- 5.2 to 11.0 +/- 5.4 mmHg (36 points, 141 FDS). The average arteriocentral venous blood pressure difference remained at 13.2 +/- 6.2 mmHg. Although it slowly decreased, the pressure difference persisted even with FDSs lasting 20 s. Lack of true equilibrium pressure could possibly be due to a waterfall mechanism. However, waterfalls were identified neither between the left ventricle and large arteries nor at the level of the diaphragm in supine patients. We therefore suggest that static filling pressures/mean circulatory pressures can only be directly assessed if the time after termination of cardiac pumping is adequate, i.e., >20 s. For humans, such times are beyond ethical options. PMID- 12907427 TI - Effects of hypoxia-reoxygenation on rat blood-brain barrier permeability and tight junctional protein expression. AB - Cerebral microvessel endothelial cells that form the blood-brain barrier (BBB) have tight junctions (TJs) that are critical for maintaining brain homeostasis. The effects of initial reoxygenation after a hypoxic insult (H/R) on functional and molecular properties of the BBB and TJs remain unclear. In situ brain perfusion and Western blot analyses were performed to assess in vivo BBB integrity on reoxygenation after a hypoxic insult of 6% O2 for 1 h. Model conditions [blood pressure, blood gas chemistries, cerebral blood flow (CBF), and brain ATP concentration] were also assessed to ensure consistent levels and criteria for insult. In situ brain perfusion revealed that initial reoxygenation (10 min) significantly increased the uptake of [14C]sucrose into brain parenchyma. Capillary depletion and CBF analyses indicated the perturbations were due to increased paracellular permeability rather than vascular volume changes. Hypoxia with reoxygenation (10 min) produced an increase in BBB permeability with associated alterations in tight junctional protein expression. These results suggest that H/R leads to reorganization of TJs and increased paracellular diffusion at the BBB, which is not a result of increased CBF, vascular volume change, or endothelial uptake of marker. Additionally, the tight junctional protein occludin had a shift in bands that correlated with functional changes (i.e., increased permeability) without significant change in expression of claudin-3, zonula occludens-1, or actin. H/R-induced changes in the BBB may result in edema and/or associated pathological outcomes. PMID- 12907429 TI - Myocardial contractility by strain echocardiography: comparison with physiological measurements in an in vitro model. AB - Strain echocardiography (SE) provides the rate and extent of myocardial segment shortening and lengthening. Thus we hypothesized that SE will noninvasively provide estimates of shortening velocity (SV) and length change (delta L). We compared SE-derived strain rate (SR) and strain (epsilon) to force/length transducer-derived SV and percent delta L in isolated muscle strips at multiple load levels and under varying conditions. Electrically stimulated contractions in left ventricular muscle strips (n = 20) were simultaneously recorded with a force/length transducer (to measure SV and percent delta L) and an ultrasound transducer (to measure SR and epsilon). Measurements were made at baseline, after inotropic stimulation, and during hypoxia at multiple load clamp levels (simulating multiple levels of afterload). Despite a difference in absolute numbers, there was a close correlation between SR and SV at baseline (R2 = 0.95), with dobutamine treatment (R2 = 0.99), and during hypoxia (R2 = 0.99). SR was load dependent at baseline (r = 0.98), with dobutamine treatment (r = 0.99), and during hypoxia (r = 0.92). Similarly, there was a close correlation between epsilon and delta L at baseline (R2 = 0.99), with dobutamine treatment (R2 = 0.96), and during hypoxia (R2 = 0.87). Percent epsilon was load dependent at baseline (r = 0.98), with dobutamine treatment (r = 0.98), and during hypoxia (r = 0.94). Bland-Altman analysis revealed a systematic overestimation of SV by SE derived SR at baseline and with dobutamine treatment. There was no bias with SR measurements during hypoxia or with epsilon measurements. SE closely tracks standard physiological parameters of regional contractile function, such as SV and delta L, under conditions of varying afterload. PMID- 12907430 TI - Demonstration of a functional apical sodium hydrogen exchanger in isolated rat gastric glands. AB - Previous studies have shown that gastric glands express at least sodium-hydrogen exchanger (NHE) isoforms 1-4. Our aim was to study NHE-3 localization in rat parietal cells and to investigate the functional activity of an apical membrane NHE-3 isoform in parietal cells of rats. Western blot analysis and immunohistochemistry showed expression of NHE-3 in rat stomach colocalizing the protein in parietal cells together with the beta-subunit of the H(+)-K(+)-ATPase. Functional studies in luminally perfused gastric glands demonstrated the presence of an apical NHE isoform sensitive to low concentrations of 5-ethylisopropyl amiloride (EIPA). Intracellular pH measurements in parietal cells conducted in omeprazole-pretreated superfused gastric glands showed an Na+-dependent proton extrusion pathway that was inhibited both by low concentrations of EIPA and by the NHE-3 specific inhibitor S3226. This pathway for proton extrusion had a higher activity in resting glands and was inhibited on stimulation of histamine induced H(+)-K(+)-ATPase proton extrusion. We conclude that the NHE-3 isoform located on the apical membrane of parietal cells offers an additional pathway for proton secretion under resting conditions. Furthermore, the gastric NHE-3 appears to work under resting conditions and inactivates during periods of H(+)-K(+) ATPase activity. PMID- 12907431 TI - Expression of enteropeptidase in differentiated enterocytes, goblet cells, and the tumor cells in human duodenum. AB - Enteropeptidase (EP) is a serine proteinase and activates trypsinogen to trypsin, thus playing an important role in food digestion. Nevertheless, the localization of EP is still controversial, likely due to a lack of studies using specific antibodies against EP. The aim of this study was to define cellular localization of EP in human duodenum and expression in tumor cells at the duodenal region. Immunohistochemical staining for resected tissues was performed with two antibodies against recombinant EP light and heavy chains, respectively. In situ hybridization was done with two RNA probes that include either the light or the heavy chain sequences of proEP, respectively. The two antibodies reacted with enterocytes, accentuated on the brush border, and goblet cells, with increasing intensity from the bottom of crypts to the top of villi. Paneth cells, neuroendocrine cells, Brunner's glands, lymphocytes, smooth muscle, or connective tissue did not react with the antibodies. The two RNA probes detected EP mRNA expression only in enterocytes and goblet cells. EP is produced in enterocytes and goblet cells, and the localization on the brush border of the cells is reasonable for the physiological activation of digestive enzymes. Interestingly, the antibodies reacted with tumor cells in duodenal polyps and adenocarcinoma at the duodenum but not in Brunner's gland adenoma. EP seems to be a marker of differentiated enterocytes and goblet cells, which suggests the existence of a common progenitor of these cells. Furthermore, EP may be a useful marker of tumor cells originating from these cells. PMID- 12907432 TI - Mixture distribution analysis of phenotypic markers reflecting HFE gene mutations. AB - The goal of this study was to determine whether statistical modeling of population data for a phenotypic marker could reflect a major locus gene defect. Identifying mutations in the HFE gene makes it possible to assess the association between transferrin saturation (TS) subpopulations and HFE mutations. Data were analyzed from 27 895 white patients who attended a health appraisal clinic and who had TS and common mutations of HFE determined. Mixture distribution modeling of TS was performed, and the proportion of HFE mutations in TS subpopulations was assessed on a probability basis. Three subpopulations of TS were identified, consistent with Hardy-Weinberg conditions for major locus effects. For men, 72% of the subpopulation with the highest mean TS had HFE gene mutations; they were primarily homozygotes or compound heterozygotes. Seventy-three percent of the subpopulation with moderate mean TS also had HFE gene mutations; they were predominantly simple heterozygotes. Sixty-seven percent of the subpopulation with the lowest mean TS were wild-type homozygotes. Similar results were observed for women. These results suggest that statistical modeling of population clinical laboratory test data can reveal the influence of a major locus gene defect and perhaps can be applied to other aspects of body metabolism than iron. PMID- 12907433 TI - Kit regulatory elements required for expression in developing hematopoietic and germ cell lineages. AB - The Kit (White) gene encodes the transmembrane receptor of stem cell factor/Kit ligand (KL) and is essential for the normal development/maintenance of pluripotent primordial germ cells (PGCs), hematopoietic stem cells (HSCs), melanoblasts, and some of their descendants. The molecular basis for the transcriptional regulation of Kit during development of these important cell types is unknown. We investigated Kit regulation in hematopoietic cells and PGCs. We identified 6 DNase I hypersensitive sites (HS1-HS6) within the promoter and first intron of the mouse Kit gene and developed mouse lines expressing transgenic green fluorescent protein (GFP) under the control of these regulatory elements. A construct driven by the Kit promoter and including all 6 HS sites is highly expressed during mouse development in Kit+ cells including PGCs and hematopoietic progenitors (erythroid blast-forming units and mixed colony-forming units). In contrast, the Kit promoter alone (comprising HS1) is sufficient to drive low-level GFP expression in PGCs, but unable to function in hematopoietic cells. Hematopoietic expression further requires the addition of the intronproximal HS2 fragment; HS2 also greatly potentiates the activity in PGCs. Thus, HS2 acts as an enhancer integrating transcriptional signals common to 2 developmentally unrelated stem cell/progenitor lineages. Optimal hematopoietic expression further requires HS3-HS6. PMID- 12907434 TI - Metalloproteinase inhibitors improve the recovery and hemostatic function of in vitro-aged or -injured mouse platelets. AB - Platelet transfusions are a crucial component of support for patients with severe thrombocytopenia. Storage of platelet concentrates, however, is associated with a reduction in platelet posttransfusion recovery and hemostatic function. In this study, we established a model of mitochondrial injury that resembles platelet storage lesion. Mitochondrial injury, provoked by incubation of platelets with carbonyl cyanide m-chlorophenylhydrazone (CCCP), led to reduced posttransfusion recovery in mice, an effect that directly correlated with the duration of treatment. Damaged platelets were characterized by shape change, disruption of membrane asymmetry, surface expression of P-selectin, and profound proteolysis of GPIbalpha. Using our model, we identified a key role for endogenous metalloproteinase(s) in platelet clearance, as their inhibition markedly improved posttransfusion recovery of both the mitochondria-injured and in vitro-aged mouse platelets. Metalloproteinase inhibition also prevented proteolysis of GPIbalpha on damaged platelets, thereby improving the hemostatic function of these cells in vivo. We propose that inhibition of metalloproteinase activity during storage could significantly improve the effectiveness of platelet transfusions. Surface expression of GPIbalpha might be a powerful marker to determine the quality of platelet concentrates, because it reflects metalloproteinase activity in vitro. PMID- 12907435 TI - Role of Ras signaling in erythroid differentiation of mouse fetal liver cells: functional analysis by a flow cytometry-based novel culture system. AB - Ras signaling plays an important role in erythropoiesis. Its function has been extensively studied in erythroid and nonerythroid cell lines as well as in primary erythroblasts, but inconclusive results using conventional erythroid colony-forming unit (CFU-E) assays have been obtained concerning the role of Ras signaling in erythroid differentiation. Here we describe a novel culture system that supports terminal fetal liver erythroblast proliferation and differentiation and that closely recapitulates erythroid development in vivo. Erythroid differentiation is monitored step by step and quantitatively by a flow cytometry analysis; this analysis distinguishes CD71 and TER119 double-stained erythroblasts into different stages of differentiation. To study the role of Ras signaling in erythroid differentiation, different H-ras proteins were expressed in CFU-E progenitors and early erythroblasts with the use of a bicistronic retroviral system, and their effects on CFU-E colony formation and erythroid differentiation were analyzed. Only oncogenic H-ras, not dominant-negative H-ras, reduced CFU-E colony formation. Analysis of infected erythroblasts in our newly developed system showed that oncogenic H-ras blocks terminal erythroid differentiation, but not through promoting apoptosis of terminally differentiated erythroid cells. Rather, oncogenic H-ras promotes abnormal proliferation of CFU-E progenitors and early erythroblasts and supports their erythropoietin (Epo) independent growth. PMID- 12907436 TI - HTLV-1 Rex is required for viral spread and persistence in vivo but is dispensable for cellular immortalization in vitro. AB - Human T-cell leukemia virus type 1 (HTLV-1) is associated with leukemia/lymphoma and neurologic disorders. Although the viral transcriptional activator Tax is the critical viral oncoprotein, Rex, which regulates the expression of the viral structural and enzymatic genes, is essential for efficient viral replication. Herein, we investigate the contribution of Rex in HTLV-1 immortalization of primary T cells in vitro and viral survival in an infectious rabbit animal model. A Rex-deficient HTLV-1 (HTLVRex-) was constructed and characterized for viral gene expression, protein production, and immortalization capacity. Cells transiently transfected with the HTLVRex- proviral clone produced low detectable levels of p19 Gag. 729HTLVRex- stable transfectants produced functional Tax, but undetectable levels of Rex or p19 Gag. Coculture of irradiated 729HTLVRex- cells with peripheral blood mononuclear cells (PBMCs) resulted in sustained interleukin 2 (IL-2)-dependent growth of primary T lymphocytes. These cells carried the HTLVRex- genome and expressed tax/rex mRNA but produced no detectable Rex or p19 Gag. Rabbits inoculated with irradiated 729HTLVRex- cells or 729HTLVRex- cells transiently transfected with a Rex cDNA expression plasmid failed to become persistently infected or mount a detectable antibody response to the viral gene products. Together, our results provide the first direct evidence that Rex and its function to modulate viral gene expression and virion production is not required for in vitro immortalization by HTLV-1. However, Rex is critical for efficient infection of cells and persistence in vivo. PMID- 12907437 TI - Alpha5beta1 integrin as a cellular coreceptor for human parvovirus B19: requirement of functional activation of beta1 integrin for viral entry. AB - Replication of the pathogenic human parvovirus B19 is restricted to erythroid progenitor cells. Although blood group P antigen has been reported to be the cell surface receptor for parvovirus B19, a number of nonerythroid cells, which express P antigen, are not permissive for parvovirus B19 infection. We have documented that P antigen is necessary for parvovirus B19 binding but not sufficient for virus entry into cells. To test whether parvovirus B19 utilizes a cell surface coreceptor for entry, we used human erythroleukemia cells (K562), which allow parvovirus B19 binding but not entry. We report here that upon treatment with phorbol esters, K562 cells become adherent and permissive for parvovirus B19 entry, which is mediated by alpha 5 beta 1 integrins, but only in their high-affinity conformation. Mature human red blood cells (RBCs), which express high levels of P antigen, but not alpha 5 beta 1 integrins, bind parvovirus B19 but do not allow viral entry. In contrast, primary human erythroid progenitor cells express high levels of both P antigen and alpha 5 beta 1 integrins and allow beta1 integrin-mediated entry of parvovirus B19. Thus, in a natural course of infection, RBCs are likely exploited for a highly efficient systemic dissemination of parvovirus B19. PMID- 12907438 TI - Coagulation, inflammation, and apoptosis: different roles for protein S and the protein S-C4b binding protein complex. AB - Protein S (PS) has an established role as an important cofactor to activated protein C (APC) in the degradation of coagulation cofactors Va and VIIIa. This anticoagulant role is evident from the consequences of its deficiency, when there is an increased risk of venous thromboembolism. In human plasma, PS circulates approximately 40% as free PS (FPS) and 60% in complex with C4b-binding protein (C4BP). Formation of this complex results in loss of PS cofactor function, and C4BP can then modulate the anticoagulant activity of APC. It had long been predicted that the complex could act as a bridge between coagulation and inflammation due to the involvement of C4BP in regulating complement activation. This prediction was recently supported by the demonstration of binding of the PS C4BP complex to apoptotic cells. This review aims to summarize recent findings on the structure and functions of PS, the basis and importance of its deficiency, its interaction with C4BP, and the possible physiologic and pathologic importance of the PS-C4BP interaction. PMID- 12907439 TI - Heparin-induced substrate behavior of antithrombin Cambridge II. AB - Cambridge II (A384S) is a highly prevalent antithrombin variant in the British population (1.14 per 1000) and predisposes carriers to a mild but significant increased risk of thrombosis. To determine if the association of Cambridge II with thrombophilia is due to a perturbation of the antithrombin inhibitory mechanism, we expressed and characterized the variant. Antithrombin Cambridge II was found to be normal in its affinity for heparin, its ability to form sodium dodecyl sulfate-stable complexes with factor Xa and thrombin, and its uncatalyzed stoichiometries and rates of inhibition. However, in the presence of full-length heparin there was a 3- and 7-fold increase in stoichiometry of inhibition of factor Xa and thrombin. The stoichiometries were not affected by pentasaccharides, indicating that the inhibitory mechanism of antithrombin Cambridge II is perturbed only in the presence of a bridging glycosaminoglycan. Thus, the vascular localization of antithrombin Cambridge II would render the carrier slightly thrombophilic. The high occurrence of this mutation and its possible propagation from a few founders suggests an evolutionary advantage, perhaps in decreasing postpartum bleeding. PMID- 12907440 TI - Three cycles of adriamycin, bleomycin, vinblastine, and dacarbazine (ABVD) or epirubicin, bleomycin, vinblastine, and methotrexate (EBVM) plus extended field radiation therapy in early and intermediate Hodgkin disease: 10-year results of a randomized trial. AB - From 1990 to 1996, a total of 386 adult patients with early/intermediate Hodgkin disease (HD) were randomly assigned to receive 3 cycles of adriamycin, bleomycin, vinblastine, dacarbazine (an alkylating agent), and methylprednisolone (ABVDm, arm A) or epirubicin, bleomycin, vinblastine, methotrexate, and methylprednisolone (EBVMm, arm E), a combination without alkylating agent. Responding patients received extended field radiation therapy (RT). Postchemotherapy complete remission and 10-year freedom from progression rates were higher in arm A (79.5% and 91.4%) than in arm E (70.4%, P =.04, and 80%, P <.002). HD mortality (HDM), treatment-related mortality (TRM), and overall survival (OS) were similar in both arms (A, 2.1%, 7.5%, and 90.4%; B, 3.9%, 5.5%, and 90.3%). However TRM and OS rates were lower in patients aged 40 years or older (P <.005), reflecting the increasing incidence of background fatal events with increasing age. Finally, event-free survival (EFS) was higher in arm A (84.6%) than in arm E (74.9%, P <.02). In patients aged younger than 40 years in arm A (74%), 10-year EFS and OS rates were 88.9% and 95.4% with HDM and TRM rates as low as 0.7% and 3%. Three courses of ABVDm plus RT are the best available option for treating early or intermediate HD. PMID- 12907441 TI - Hepatosplenic gammadelta T-cell lymphoma is a rare clinicopathologic entity with poor outcome: report on a series of 21 patients. AB - We report on the characteristics of 21 patients with hepatosplenic gammadelta T cell lymphoma (HSgammadeltaTCL), an entity recognized since 1994 in the Revised European American Lymphoma (REAL) classification. Median age was 34 years. Patients had splenomegaly (n = 21), hepatomegaly (n = 15), and thrombocytopenia (n = 20). Histopathologic findings were homogeneous and showed the presence of medium-sized lymphoma cells within the sinusoids of splenic red pulp, liver, and bone marrow. Marrow involvement was usually mild but could be demonstrated by phenotyping in all patients. Cells were CD3+CD5-, expressed the gammadelta T-cell receptor, and had a nonactivated cytotoxic cell phenotype (TIA-1+, granzyme B-). Most patients were CD4-/CD8- (16 of 18); CD56+ (15 of 18), expressed the Vdelta1epitope (Vd1+/Vd2-/Vd3-) (9 of 12); and were negative for Epstein-Barr virus (EBV) (18 of 20). Isochromosome arm 7q was documented in 9 of 13 patients. Eight patients had previously undergone kidney transplantation or had a history of systemic lupus, Hodgkin disease, or malaria. Prognosis was poor; median survival time was 16 months, and all but 2 patients ultimately died despite consolidative or salvage high-dose therapy. In conclusion, HSgammadeltaTCL is a disease with distinctive clinical, histopathologic, and phenotypic characteristics. Bone marrow biopsy with combined phenotyping is sufficient for diagnosis, and splenectomy is therefore unwarranted. Current treatment modalities appear to be ineffective in most patients. PMID- 12907442 TI - Molecular histogenesis of posttransplantation lymphoproliferative disorders. AB - Posttransplantation lymphoproliferative disorders (PTLDs) represent a serious complication of solid organ transplantation. This study assessed the molecular histogenesis of 52 B-cell monoclonal PTLDs, including 12 polymorphic PTLDs (P PTLDs), 36 diffuse large B-cell lymphomas (DLBCLs), and 4 Burkitt/Burkitt-like lymphomas (BL/BLLs). Somatic hypermutation (SHM) of immunoglobulin variable (IgV) genes documented that most monoclonal B-cell PTLDs (75% P-PTLDs, 91.3% DLBCLs, 100% BL/BLLs) derive from germinal center (GC)-experienced B cells. B-cell lymphoma 6 (BCL6) mutations occurred in 25% P-PTLDs, 60.6% DLBCLs, and 75.0% BL/BLLs. A first histogenetic category of PTLDs (31.2% DLBCLs) express the BCL6+/multiple myeloma oncogene-1 protein (MUM1-/+)/CD138- profile and mimic B cells experiencing the GC reaction, as also suggested by ongoing SHM in a fraction of these cases. A second subset of PTLDs (66.7% P-PTLDs and 31.2% DLBCLs) display the BCL6-/MUM1+/CD138- phenotype and mimic B cells that have concluded the GC reaction. A third histogenetic category of PTLDs (25.0% P-PTLDs and 31.2% DLBCLs) shows the BCL6-/MUM1+/CD138+ profile, consistent with preterminally differentiated post-GC B cells. Crippling mutations of IgV heavy chain (IgVH) and/or IgV light chain (IgVL) genes, leading to sterile rearrangements and normally preventing cell survival, occur in 4 DLBCLs and 1 BL/BLL that may have been rescued from apoptosis through expression of Epstein Barr virus (EBV)-encoded latent membrane protein 1 (LMP1). Overall, the histogenetic diversity of monoclonal B-cell PTLDs may help define biologically homogeneous categories of the disease. PMID- 12907443 TI - Effects of 5-aza-2'-deoxycytidine on fetal hemoglobin levels, red cell adhesion, and hematopoietic differentiation in patients with sickle cell disease. AB - Fetal hemoglobin (HbF) decreases polymerization of sickle hemoglobin (HbS) and improves outcomes in sickle cell disease (SSD). Therefore, a therapeutic goal in SSD is pharmacologic reactivation of HbF. Silencing of the gamma-globin (HbF) gene is associated with DNA methylation. The cytosine analog 5-aza-2' deoxycytidine (decitabine) hypomethylates DNA by inhibiting DNA methyltransferase. We examined if subcutaneous decitabine could increase HbF levels and improve SSD pathophysiology without cytotoxicity. Eight symptomatic SSD patients resistant or intolerant of standard treatment with hydroxyurea received decitabine 0.2 mg/kg subcutaneously 1 to 3 times per week in 2 cycles of 6-week duration. Treatment decreased neutrophils and increased mean HbF (6.5% to 20.4%, P <.0001) and mean total hemoglobin (76 to 96 g/L [7.6 to 9.6 g/dL], P <.001). Features of vaso-occlusive crisis pathophysiology such as red cell adhesion, endothelial damage, and coagulation pathway activity significantly improved. gamma-Globin gene promoter methylation decreased, and platelets and the proportion of megakaryocytes and erythroid cells in the marrow increased without a decrease in marrow cellularity, consistent with a DNA hypomethylating, noncytotoxic mechanism of action. Weekly subcutaneous decitabine produces cumulative increases in HbF and total hemoglobin through a noncytotoxic mechanism of action. Chronic dosing and sustained increases in hemoglobin F and total hemoglobin levels may be possible. Further studies in SSD and thalassemia are indicated. PMID- 12907444 TI - Lack of evidence for functional ADP-activated human P2X1 receptors supports a role for ATP during hemostasis and thrombosis. AB - Purine nucleotides acting through P2 receptors play key roles in platelet signaling. The P2X1 receptor is an adenosine triphosphate (ATP)-gated ion channel that mediates a rapid calcium influx signal, but can also synergize with subsequent adenosine diphosphate (ADP)-evoked P2Y1 receptor-mediated responses and thus may contribute to platelet activation during hemostasis. Recent studies have shown that P2X1 receptors contribute to the formation of platelet thrombi, particularly under conditions of high shear stress. Based on intracellular Ca2+ measurements a previous report has suggested that a splice variant of the P2X1 receptor, P2X1del, is expressed in platelets and, in contrast to the full-length P2X1WT receptor, is activated by ADP. In the present study we show that the P2X1del receptor fails to form functional ion channels and is below the limit of detection in human platelets. Furthermore, ADP does not contribute to the rapid ionotropic P2X receptor-mediated response in platelets. These results support the notion that ATP is the principal physiologic agonist at P2X1 receptors and that it plays a role in the activation of platelets. PMID- 12907445 TI - CD25 expression on donor CD4+ or CD8+ T cells is associated with an increased risk for graft-versus-host disease after HLA-identical stem cell transplantation in humans. AB - Graft-versus-host disease (GVHD) occurs in an unpredictable fashion after 30% to 50% of matched-related transplantations. The presence of increased frequencies of CD4(+)CD25(+) regulatory T cells in donor grafts has been shown to ameliorate GVHD after allogeneic transplantation in murine models. To determine whether a similar relationship exists in humans, we quantitated the coexpression of CD25 on CD4(+) and CD8(+) T cells within 60 donor grafts infused into matched siblings and examined GVHD incidence in the respective recipients. Recipients in whom GVHD developed received donor grafts containing significantly higher frequencies of CD4(+) T cells coexpressing CD25 than those who did not (median, 9.26% vs 2.22%; P =.004). Frequencies of donor graft CD8(+) T cells coexpressing CD25 were also higher (0.65% vs 0.14%; P =.002). Furthermore, transplant recipients who received grafts containing fewer CD4(+)CD25(+) and CD8(+)CD25(+) T cells were less likely to acquire acute GVHD, even though these donor-recipient pairs were similar to others with respect to relevant clinical variables. These data suggest that the coexpression of CD4 and CD25 may be insufficient to identify regulatory T cells in humans and that increased frequencies and numbers of CD25(+) T cells in donor grafts is associated with GVHD in transplant recipients. PMID- 12907446 TI - Rituximab as adjuvant to high-dose therapy and autologous hematopoietic cell transplantation for aggressive non-Hodgkin lymphoma. AB - Based on the favorable safety profile and the independent activity of rituximab in B-cell lymphoma, we evaluated its efficacy and toxicity after high-dose therapy (HDT) and autologous hematopoietic cell transplantation (HCT). Thirty five patients with diffuse large cell (25 patients), mantle cell (3 patients), transformed (3 patients), or other (4 patients) subtypes of B-cell lymphoma received HDT followed by a purged autologous graft. The rituximab schedule was 4 weekly infusions (375 mg/m(2)) starting at day 42 after HCT and, for patients 5 to 35, a second 4-week course 6 months after HCT. All planned therapy was completed in 29 patients. With 30 months' median follow-up, the 2-year event-free survival (EFS) rate was 83% and the overall survival (OS) rate was 88%. For 21 patients with relapsed or refractory large cell lymphoma, the EFS rate was 81% and the OS rate was 85%. Grades 3 to 4 neutropenia occurred in 19 (54%) patients. A prospective study of immune reconstitution included measurements of lymphocyte subsets, immunoglobulins, and response to vaccination. Serious infections were not observed despite delayed B-cell recovery in all patients and suppressed immunoglobulin G (IgG) levels and low pneumococcus antibody titers in a subset. Rituximab after HDT and HCT is feasible, and these phase 2 data support the current US Intergroup phase 3 trial in recurrent/refractory diffuse large cell lymphoma. PMID- 12907447 TI - Late-onset noninfectious pulmonary complications after allogeneic stem cell transplantation are significantly associated with chronic graft-versus-host disease and with the graft-versus-leukemia effect. AB - Late-onset noninfectious pulmonary complications (LONIPCs) occurring beyond 3 months after allogeneic stem cell transplantation (allo-SCT) have become recognized as life-threatening complications, and they reduce the recipient's quality of life. However, the pathogenesis and optimal treatment for LONIPCs are still unclear. In this study, we retrospectively analyzed the incidence and outcome of LONIPCs among allo-SCT recipients. Between October 1993 and September 2001, 96 patients underwent allo-SCT and 76 patients who survived and were free of disease for more than 3 months after SCT were enrolled. Among the 76 patients, 18 patients (23.7%) developed LONIPCs at a median interval of 227 days after allo SCT (range, 91-1105 days). The patients with LONIPCs were subclassified into those with bronchiolitis obliterans (BO) (6 patients), with interstitial pneumonia (IP) (11 patients), or with both BO and IP (1 patient). The presence of extensive chronic graft-versus-host disease (GVHD) was significantly associated with the development of LONIPCs (P =.0008). Liver or skin involvement in chronic GVHD was not associated, but sicca syndrome was significantly associated with the development of LONIPCs (P <.0001). Most of the IP patients (58.3%) responded well to immunosuppressive treatment, while BO patients did not respond to the therapy. Eight of the 18 patients with LONIPCs died. The major cause of death was respiratory failure (62.5%). The relapse rate of primary malignant disease in the LONIPC patients was significantly lower than that of non-LONIPC patients (1 of 17 [5.9%] versus 16 of 52 [30.8%]; P =.0387). These results indicate that the development of LONIPCs was strongly associated with chronic GVHD and especially with sicca syndrome and the graft-versus-leukemia (GVL) effect. PMID- 12907448 TI - Erythroid pyrimidine 5'-nucleotidase: cloning, developmental expression, and regulation by cAMP and in vivo hypoxia. AB - A characteristic process of terminal erythroid differentiation is the degradation of ribosomal RNA into mononucleotides. The pyrimidine mononucleotides can be dephosphorylated by pyrimidine 5'-nucleotidase (P5N-I). In humans, a lack of this enzyme causes hemolytic anemia with ribosomal structures and trinucleotides retained in the red blood cells (RBCs). Although the protein/nucleotide sequence of P5N-I is known in mammals, the onset and regulation of P5N-I during erythroid maturation is unknown. However, in circulating chicken embryonic RBCs, the enzyme is induced together with carbonic anhydrase (CAII) and 2,3-bisphosphoglycerate (2,3-BPG) by norepinephrine (NE) and adenosine, which are released by the embryo under hypoxic conditions. Here, we present the chicken P5N-I sequence and the gene expression of P5N-I during RBC maturation; the profile of gene expression follows the enzyme activity with a rise between days 13 and 16 of embryonic development. The p5n-I expression is induced (1) in definitive but not primitive RBCs by stimulation of beta-adrenergic/adenosine receptors, and (2) in definitive RBCs by hypoxic incubation of the chicken embryo. Since embryonic RBCs increase their hemoglobin-oxygen affinity by degradation of nucleotides such as uridine triphosphate (UTP) and cytidine triphosphate (CTP), the induction of p5n-I expression can be seen as an adaptive response to hypoxia. PMID- 12907449 TI - Chemokine stimulation of human peripheral blood T lymphocytes induces rapid dephosphorylation of ERM proteins, which facilitates loss of microvilli and polarization. AB - Lymphocyte microvilli mediate initial rolling-adhesion along endothelium but are lost during transmigration from circulation to tissue. However, the mechanism for resorption of lymphocyte microvilli remains unexplored. We show that chemokine stimulation of human peripheral blood T (PBT) cells is sufficient to induce rapid resorption of microvilli. Microvilli in other cells are regulated by ezrin/radixin/moesin (ERM) proteins, which link the plasma membrane to the cortical F-actin cytoskeleton; maintenance of these linkages requires ERM activation, reflected by phosphorylation at a specific carboxy-terminal threonine residue. Carboxyphosphorylated-ERM (cpERM) proteins in resting PBT cells show a punctate peripheral distribution consistent with localization to microvilli. cpERM dephosphorylation begins within seconds of stimulation by chemokines (stromal derived factor 1 alpha [SDF-1 alpha] or secondary lymphoid tissue cytokine), and ERM proteins lose their punctate distribution with kinetics paralleling the loss of microvilli. The cpERM proteins are preferentially associated with the cytoskeleton at rest and this association is lost with chemokine-induced dephosphorylation. Transfection studies show that a dominant negative ERM construct destroys microvilli, whereas a construct mimicking cpERM facilitates formation of microvilli, retards chemokine-induced loss of microvilli, and markedly impairs chemokine-induced polarization. Thus, chemokine induces rapid dephosphorylation and inactivation of cpERM, which may in turn facilitate 2 aspects of cytoskeletal reorganization involved in lymphocyte recruitment: loss of microvilli and polarization. PMID- 12907450 TI - Suppressor of cytokine signaling 1 attenuates IL-15 receptor signaling in CD8+ thymocytes. AB - SOCS1-/- mice die prematurely of increased interferon-gamma (IFNgamma) signaling with severe thymic atrophy and accelerated maturation of T cells. However, it was unclear whether the thymic defects were caused by SOCS1 deficiency or by increased IFNgamma signaling. Using SOCS1-/- IFNgamma-/- mice, we show in this study that SOCS1 deficiency skews thymocyte development toward CD8 lineage independently of IFNgamma. Fetal thymic organ cultures and intrathymic transfer of CD4-CD8- precursors into Rag1-/- mice show that the lineage skewing in SOCS1-/ mice is a T-cell autonomous defect. Interestingly, SOCS1 is not required for attenuating interleukin-7 (IL-7) signaling at the CD4-CD8- stage but is essential for regulating IL-15 and IL-2 signaling in CD8+ thymocytes. IL-15 selectively stimulates SOCS1-/- CD8+ thymocytes, inducing sustained signal transducer and activator of transcription 5 (STAT5) phosphorylation and massive proliferation. IL-15 also strongly up-regulates Bcl-xL and CD44 in CD8+ thymocytes lacking SOCS1. The SOCS1 gene is induced in CD4+ thymocytes by gammac cytokines, whereas CD8+ thymocytes constitutively express SOCS1 mRNA even in the absence of cytokine stimulation. Because many different cell types express IL-15, our results strongly suggest that SOCS1 functions as an indispensable attenuator of IL-15 receptor signaling in developing CD8+ thymocytes. PMID- 12907451 TI - Human dendritic cells express neuronal Eph receptor tyrosine kinases: role of EphA2 in regulating adhesion to fibronectin. AB - Eph receptor tyrosine kinases and their ligands, the ephrins, have been primarily described in the nervous system for their roles in axon guidance, development, and cell intermingling. Here we address whether Eph receptors may also regulate dendritic cell (DC) trafficking. Reverse transcription-polymerase chain reaction (RT-PCR) analysis showed that DCs derived from CD34+ progenitors, but not from monocytes, expressed several receptors, in particular EphA2, EphA4, EphA7, EphB1, and EphB3 mRNA. EphB3 was specifically expressed by Langerhans cells, and EphA2 and EphA7 were expressed by both Langerhans- and interstitial-type DCs. EphA and EphB protein expression on DCs generated in vitro was confirmed by staining with ephrin-A3-Fc and ephrin-B3-Fc fusion proteins that bind to different Eph members, in particular EphA2 and EphB3. Immunostaining with anti-EphA2 antibodies demonstrated the expression of EphA2 by immature DCs and by skin Langerhans cells isolated ex vivo. Interestingly, ephrin expression was detected in epidermal keratinocytes and also in DCs. Adhesion of CD34+-derived DCs to fibronectin, but not to poly-l-lysine, was increased in the presence of ephrin-A3-Fc, a ligand of EphA2, through a beta1 integrin activation pathway. As such, EphA2/ephrin-A3 interactions may play a role in the localization and network of Langerhans cells in the epithelium and in the regulation of their trafficking. PMID- 12907452 TI - Selective expression of stromal-derived factor-1 in the capillary vascular endothelium plays a role in Kaposi sarcoma pathogenesis. AB - Kaposi sarcoma (KS), the most common neoplasm in patients with AIDS, typically presents with multiple skin lesions characterized by "spindle cells," the vast majority of which are infected with KSHV (Kaposi sarcoma herpes virus, also named HHV-8). In patients with AIDS, the presence of cell-associated KSHV DNA in blood is predictive of subsequent KS development, but the mechanisms by which circulating KSHV-infected cells contribute to AIDS-KS pathogenesis are unclear. Here, we show that the chemokine stromal-derived factor-1 (SDF-1), which is constitutively expressed by skin capillary endothelium and displayed on the endothelial cell surface in association with heparan sulfate, can trigger specific arrest of KSHV-infected cells under physiologic shear flow conditions. Moreover, in the presence of soluble SDF-1 gradients, SDF-1 expressed on the endothelial barrier can promote transendothelial migration of KSHV-infected cells. By triggering specific adhesion of circulating KSHV-infected cells and favoring their entry into the extravascular cutaneous space, endothelial cell associated SDF-1 in cutaneous capillaries may dictate the preferential occurrence of KS in the skin. PMID- 12907453 TI - JunB inhibits proliferation and transformation in B-lymphoid cells. AB - The activator protein 1 (AP-1) member JunB has recently been implicated in leukemogenesis. Here we surveyed human lymphoma samples for expression of JunB and other AP-1 members (c-Jun, c-Fos, Fra1, JunD). JunB was strongly expressed in T-cell lymphomas, but non-Hodgkin B-cell lymphomas do not or only weakly express JunB. We therefore asked whether JunB acted as a negative regulator of B-cell development, proliferation, and transformation. We used transgenic mice that expressed JunB under the control of the ubiquitin C promoter; these displayed increased JunB levels in both B- and T-lymphoid cells. JunB transgenic cells of B lymphoid, but not of T-lymphoid, origin responded poorly to mitogenic stimuli. Furthermore, JunB transgenic cells were found to be less susceptible to the transforming potential of the Abelson oncogene in vitro. In addition, overexpression of JunB partially protected transgenic mice against the oncogenic challenge in vivo. However, transformed B cells eventually escaped from the inhibitory effect of JunB: the proliferative response was similar in explanted tumor-derived cells from transgenic animals and those from wild-type controls. Our results identify JunB as a novel regulator of B-cell proliferation and transformation. PMID- 12907455 TI - Epstein-Barr Virus (EBV) LMP2A induces alterations in gene transcription similar to those observed in Reed-Sternberg cells of Hodgkin lymphoma. AB - Epstein-Barr virus (EBV) is associated with the development of a variety of malignancies, including Hodgkin lymphoma. One of the few viral transcripts expressed in EBV-positive Hodgkin/Reed-Sternberg (HRS) cells of Hodgkin lymphoma is latent membrane protein 2A (LMP2A). This viral protein blocks B-cell receptor (BCR)-signaling in vitro. Furthermore, expression of LMP2A in developing B cells in vivo induces a global down-regulation of genes necessary for proper B-cell development. In this study we have analyzed gene transcription in primary B cells from LMP2A transgenic mice, LMP2A-expressing human B-cell lines, and LMP2A positive and -negative EBV-infected lymphoblastoid cell lines (LCLs). We demonstrate that LMP2A increases the expression of genes associated with cell cycle induction and inhibition of apoptosis, alters the expression of genes involved in DNA and RNA metabolism, and decreases the expression of B-cell specific factors and genes associated with immunity. Furthermore, many alterations in gene expression induced by LMP2A are similar to those recently described in HRS cells of Hodgkin lymphoma and activated, proliferating germinal center centroblasts/centrocytes. These correlations suggest that LMP2A expression in EBV-infected B cells may lead to the induction and maintenance of an activated, proliferative state that could ultimately result in the development of Hodgkin lymphoma. PMID- 12907454 TI - A role for Rab27b in NF-E2-dependent pathways of platelet formation. AB - Megakaryocytes release platelets by reorganizing the cytoplasm into proplatelet extensions. Fundamental to this process is the need to coordinate transport of products and organelles in the appropriate abundance to nascent platelets. The importance of the Rab family of small GTPases (guanosine 5'-triphosphatases) in platelet biogenesis is revealed in gunmetal (gm/gm) mice, which show deficient Rab isoprenylation and macrothrombocytopenia with few granules and abnormal megakaryocyte morphology. Although some Rab proteins are implicated in vesicle and organelle transport along microtubules or actin, the role of any Rab protein in platelet biogenesis is unknown. The limited number of Rab proteins with defective membrane association in gm/gm megakaryocytes prominently includes Rab27a and Rab27b. Normal expression of Rab27b is especially increased with terminal megakaryocyte differentiation and dependent on nuclear factor-erythroid 2 (NF-E2), a transcription factor required for thrombopoiesis. Chromatin immunoprecipitation demonstrates recruitment of NF-E2 to the putative Rab27B promoter. Inhibition of endogenous Rab27 function in primary megakaryocytes causes severe quantitative and qualitative defects in proplatelet formation that mimic findings in gm/gm cells. Rab27b localizes to alpha and dense granules in megakaryocytes. These results establish a role for Rab27 in platelet synthesis and suggest that Rab27b in particular may coordinate proplatelet formation with granule transport, possibly by recruiting specific effector pathways. PMID- 12907456 TI - Notch signaling is necessary but not sufficient for differentiation of dendritic cells. AB - The Notch family of receptors plays an important role in regulation of cell differentiation via direct contact between hematopoietic progenitor cells (HPCs) and bone marrow stroma (BMS). However the precise contribution of Notch in dendritic cell (DC) differentiation is controversial. In 2 different experimental systems using Notch-1-null embryonic stem cells and Notch-1-deficient HPCs we have found that Notch-1 is necessary for DC differentiation. However, activation of Notch-1 and Notch-2 with cell-bound Notch ligand did not result in differentiation of mature DCs or macrophages. Instead, it caused accumulation of immature myeloid cells. Removal of feeder cells resulted in rapid differentiation of DCs and macrophages. Addition of interleukin 4 (IL-4) into the culture dramatically increased accumulation of functionally potent DCs. Lipopolysaccharide was not able to reproduce this effect. Thus, these data indicate that Notch signaling prevents differentiation of mature myeloid cells. Instead, it results in accumulation of precursors readily able to differentiate into mature DCs once the Notch signal is stopped (eg, after cell emigration from bone marrow) and in the presence of other additional differentiation signals provided by IL-4. Thus, Notch is required but not sufficient for DC differentiation. PMID- 12907457 TI - Cloning of the t(1;5)(q23;q33) in a myeloproliferative disorder associated with eosinophilia: involvement of PDGFRB and response to imatinib. AB - Eosinophilia is common in myeloproliferative disorders (MPDs) with abnormalities of chromosome band 5q31-33, including those that present with t(1;5)(q23;q33). With the development of rational drug therapy, characterization of the molecular targets for these translocations could guide treatment and affect patient survival. We cloned the t(1;5)(q23;q33) and showed that it fuses platelet-derived growth factor receptor beta (PDGFRB) to the coiled-coil domains of a novel partner protein, myomegalin. Using two-color interphase fluorescence in situ hybridization (FISH), we also demonstrated that the eosinophils are clonal in these disorders. Imatinib mesylate has recently been shown to be efficacious in MPDs with PDGFR activation. Therefore, following our molecular studies, we were able to redirect this patient's treatment. Although she had refractory and progressive disease, once imatinib was started, complete clinical and hematologic remission, as well as major cytogenetic response, was achieved. Given the therapeutic implications, our findings stress the need to aggressively investigate the molecular basis of these diseases, with emphasis on the PDGFR family. PMID- 12907458 TI - IL-10-inducible Bcl-3 negatively regulates LPS-induced TNF-alpha production in macrophages. AB - Interleukin-10 (IL-10) plays an important role in prevention of chronic inflammation in vivo. However, the molecular mechanism by which IL-10 exerts its anti-inflammatory response is poorly understood. Here, we performed a microarray analysis and identified Bcl-3 as an IL-10-inducible gene in macrophages. Lentiviral vector-mediated expression of Bcl-3 inhibited lipopolysaccharide (LPS) induced production of tumor necrosis factor alpha (TNF-alpha), but not IL-6, in macrophages. In Bcl-3-transduced and IL-10-pretreated macrophages, LPS-induced nuclear translocation of nuclear factor kappaB (NF-kappaB) p65 was not impaired. However, DNA binding by NF-kappaB p50/p65 was profoundly inhibited. Nuclear localization of Bcl-3 was associated with inhibition of LPS-induced TNF-alpha production. Overexpression of Bcl-3 suppressed activation of the TNF-alpha promoter, but not the IL-6 promoter. Bcl-3 interacted with NF-kappaB p50 and was recruited to the TNF-alpha promoter, but not the IL-6 promoter, indicating that Bcl-3 facilitates p50-mediated inhibition of TNF-alpha expression. Furthermore, Bcl-3-deficient macrophages showed defective IL-10-mediated suppression of LPS induction of TNF-alpha, but not IL-6. These findings suggest that IL-10-induced Bcl-3 is required for suppression of TNF-alpha production in macrophages. PMID- 12907459 TI - Iron loading and erythrophagocytosis increase ferroportin 1 (FPN1) expression in J774 macrophages. AB - The expression of ferroportin1 (FPN1) in reticuloendothelial macrophages supports the hypothesis that this iron-export protein participates in iron recycling from senescent erythrocytes. To gain insight into FPN1's role in macrophage iron metabolism, we examined the effect of iron status and erythrophagocytosis on FPN1 expression in J774 macrophages. Northern analysis indicated that FPN1 mRNA levels decreased with iron depletion and increased on iron loading. The iron-induced induction of FPN1 mRNA was blocked by actinomycin D, suggesting that transcriptional control was responsible for this effect. After erythrophagocytosis, FPN1 mRNA levels were also up-regulated, increasing 8-fold after 4 hours and returning to basal levels by 16 hours. Western analysis indicated corresponding increases in FPN1 protein levels, with maximal induction after 10 hours. Iron chelation suppressed FPN1 mRNA and protein induction after erythrophagocytosis, suggesting that FPN1 induction results from erythrocyte derived iron. Comparative Northern analyses of iron-related genes after erythrophagocytosis revealed a 16-fold increase in FPN1 levels after 3 hours, a 10-fold increase in heme oxygenase-1 (HO-1) after 3 hours, a 2-fold increase in natural resistance macrophage-associated protein 1 (Nramp1) levels after 6 hours, but no change in divalent metal ion transporter 1 (DMT1) levels. The rapid and strong induction of FPN1 expression after erythrophagocytosis suggests that FPN1 plays a role in iron recycling. PMID- 12907460 TI - Induction of atherosclerosis by low-fat, semisynthetic diets in LDL receptor deficient C57BL/6J and FVB/NJ mice: comparison of lesions of the aortic root, brachiocephalic artery, and whole aorta (en face measurement). AB - OBJECTIVE: A semisynthetic diet with varying amounts of cholesterol was used to achieve hypercholesterolemia and atherosclerosis in LDL receptor-deficient (LDLR /-) mice. Atherosclerotic lesions were measured as cross-sectional area at the aortic root and brachiocephalic artery and by en face analysis of aortic lesion area in 209 male and female animals on the C57BL/6J (B6.LDLR-/-) and FVB/NJ (FVB.LDLR-/-) backgrounds. METHODS AND RESULTS: The semisynthetic diet containing 4.3% fat and 0.00% or 0.02% cholesterol was sufficient to induce hypercholesterolemia (12.6+/-2.4 mmol/L) and atherosclerosis in B6.LDLR-/- mice at the aortic root (98,980+/-37 727 microm2) and brachiocephalic artery (12,039+/ 12,750 microm2) but did not produce significant lesions in the aorta measurable by the en face method. Raising dietary cholesterol to 0.15%, 0.30%, or 0.50% more than doubled plasma cholesterol levels (35.9+/-8.5 mmol/L) and resulted in significant en face lesions. It also led to a significant increase in atherosclerotic lesion area at the aortic root (547,753+/-182,151 microm2) and brachiocephalic arteries (125,666+/-59,339 microm2). Although FVB.LDLR-/- mice developed comparable cholesterol levels, they were relatively atherosclerosis resistant and had many-fold smaller lesions. CONCLUSIONS: These results should aid investigations of atherosclerosis in LDLR-/- mice by informing the selection of diet to be used and the location of lesions to be scored. PMID- 12907461 TI - Promoter but not exon 7 polymorphism of endothelial nitric oxide synthase affects training-induced correction of endothelial dysfunction. AB - Background- Polymorphisms in the promoter (T-786C) and exon 7 (G894T) of the endothelial nitric oxide synthase (eNOS) gene were shown to be associated with reduced vascular NO production or increased proteolytic cleavage of eNOS. Therefore, we aimed to determine the effects of these polymorphisms on endothelial function and endothelial response to physical exercise in patients with coronary artery disease (CAD). METHODS AND RESULTS: Sixty-seven patients were randomized to either a training or a control group. At the beginning and after 4 weeks, acetylcholine-induced changes in average peak velocity (APV) of a coronary or mammary artery were invasively assessed by Doppler velocimetry. Polymorphisms were detected by polymerase chain reaction-restriction fragment length polymorphism. At the beginning, in subjects with the wild-type (WT) variant, APV increased by 88+/-7% in response to acetylcholine. This response was significantly blunted in patients who were positive for the promoter (44+/-7%) or the exon 7 (62+/-9%) polymorphism. Four weeks of exercise training resulted in augmentation of an endothelium-dependent increase in APV by +36+/-12% in promoter polymorphism-positive patients but by +81+/-18% and +91+/-15% in WT variant- and exon 7 polymorphism-positive subjects, respectively. CONCLUSIONS: These results suggest that the presence of either one of the polymorphisms attenuates endothelium-dependent vasodilatation in CAD patients. Only the promoter polymorphism might have an adverse effect on training-induced improvement in endothelial function. PMID- 12907462 TI - Cholesterol dependence of vascular ERK1/2 activation and growth in response to stretch: role of endothelin-1. AB - OBJECTIVE: Stretch-induced growth of the vascular wall plays a role in hypertension and neointima formation. Its signal pathways involve integrins, cytoskeleton, membrane receptors, and ion channels, some of which are organized in cholesterol-rich, membrane domains such as lipid rafts or caveolae. This study tested the role of rafts/caveolae in stretch-induced vascular growth by manipulation of membrane cholesterol contents. METHODS AND RESULTS: Growth and protein synthesis were induced by mechanical stretch of rat portal veins in vitro. Sucrose gradient centrifugation showed stretch-induced tyrosine phosphorylation primarily in fractions containing caveolin-1. Disruption of membrane caveolae with use of methyl-beta-cyclodextrin (mbetacd) reduced weight gain, protein synthesis, and DNA synthesis to levels in unstretched, control veins. These effects were partially reversed by restoration of cellular cholesterol contents. Inhibited growth was associated with abolished activation of extracellular signal-regulated kinase (ERK) 1/2 in response to stretch and endothelin-1 (ET-1) but not to angiotensin II. Inhibition of ET-1 type A (ETA) receptors by RF139317 or endothelin-converting enzyme by phosphoramidone abolished stretch-induced ERK1/2 activation, which was, however, unaffected by removal of the endothelium. CONCLUSIONS: Stretch-induced growth signaling in vascular smooth muscle depends on cholesterol-rich, membrane microdomains by a mechanism involving ETA receptors that respond to endogenous ET-1 production. PMID- 12907463 TI - Arterial smooth muscle cell heterogeneity: implications for atherosclerosis and restenosis development. AB - During atheromatous plaque formation or restenosis after angioplasty, smooth muscle cells (SMCs) migrate from the media toward the intima, where they proliferate and undergo phenotypic changes. The mechanisms that regulate these phenomena and, in particular, the phenotypic modulation of intimal SMCs have been the subject of numerous studies and much debate during recent years. One view is that any SMCs present in the media could undergo phenotypic modulation. Alternatively, the seminal observation of Benditt and Benditt that human atheromatous plaques have the features of a monoclonal or an oligoclonal lesion has led to the hypothesis that a predisposed, medial SMC subpopulation could play a crucial role in the production of intimal thickening. The presence of a distinct SMC population in the arterial wall implies that under normal conditions, SMCs are phenotypically heterogeneous. The concept of SMC heterogeneity is gaining wider acceptance, as shown by the increasing number of publications on this subject. In this review, we discuss the in vitro studies that demonstrate the presence of distinct SMC subpopulations in arteries of various species, including humans. Their specific features and their regulation will be highlighted. Finally, the relevance of an atheroma-prone phenotype to intimal thickening formation will be discussed. PMID- 12907464 TI - Ethanol inhibits fibroblast growth factor-induced proliferation of aortic smooth muscle cells. AB - OBJECTIVE: Epidemiological studies have demonstrated that moderate alcohol consumption reduces mortality associated with coronary artery disease. The protective effect is correlated with the amount of ethanol consumed but is unrelated to the form of alcoholic beverage. Adoption of a favorable lipoprotein profile accounts for about half of the protective action of alcohol, but the remaining causative factors remain conjectural. Fibroblast growth factors (FGFs) play important roles in mediating smooth muscle cell (SMC) proliferation and migration, which are key factors in the atherosclerotic process. In the present study, we examined the effect of ethanol on FGF-mediated SMC growth and signaling. METHODS AND RESULTS: Pharmacologically relevant concentrations of ethanol inhibited the proliferation of a rat aortic SMC line (SV40LT-SMCs) in response to FGF1 and FGF2. Human aortic SMC growth was similarly inhibited by ethanol. Transition into the G2/M phase was specifically affected. FGF-mediated phosphorylation of p42/p44 mitogen-activated protein kinase (MAPK) c-Raf, MAP kinase kinase kinase, MEK1/2 MAP kinase, kinase, stress-activated protein kinase/c-Jun-NH2-terminal kinase, and p38 MAPK were variably reduced by ethanol. The inhibition of intracellular signaling by ethanol was correlated with inhibition of FGF receptor autophosphorylation. By contrast, neither epidermal growth factor receptor autophosphorylation nor epidermal growth factor-mediated p42/p44 MAPK activation was affected by ethanol. CONCLUSIONS: The findings identify the FGF receptor as an inhibitory target for ethanol, which could account in part for the inhibitory actions of ethanol on SMC proliferation observed in vivo. PMID- 12907465 TI - Thiazolidinediones and blood lipids in type 2 diabetes. AB - We evaluated study population characteristics and treatment effects on blood lipids between studies in which either rosiglitazone (RSG) or pioglitazone (PIO) was investigated in patients with type 2 diabetes. We performed a summary analysis of all published double-blind, placebo-controlled studies with RSG (4 and 8 mg/d) and PIO (15, 30, and 45 mg/d). Data were analyzed by the random effects model. Nineteen trials met our inclusion criteria, yielding 5304 patients, 3236 in studies with RSG and 2068 in studies with PIO. Subjects treated with PIO were more obese and showed more pronounced hyperglycemia and dyslipidemia (increased triglycerides and decreased HDL cholesterol) at baseline than did subjects treated with RSG. By weighted linear-regression analysis, studies with PIO showed greater beneficial effects on triglycerides, total cholesterol, and LDL cholesterol, after adjustment for the respective lipid levels at baseline. RSG 8 mg/d showed greater increases in total cholesterol and LDL cholesterol than did RSG 4 mg/d. PIO 30 mg/d showed greater reductions in triglycerides than did PIO 15 mg/d. Studies conducted with PIO showed more beneficial effects on blood lipids, but also different study population characteristics in comparison with studies conducted with RSG. Differences in both pharmacologic properties between agents and study population characteristics are likely to have influenced the results. PMID- 12907466 TI - An in vivo experimental system to study sugar phloem unloading in ripening grape berries during water deficiency stress. AB - An in vivo experimental system-called the 'berry-cup' technique-was developed to study sugar phloem unloading and the accumulation of sugar in ripening grape berries. The berry-cup system consists of a single peeled grape berry immersed in a buffer solution in a cup prepared from a polypropylene syringe. A small cross incision (2 mm in length) is made on the stylar remnant of a berry during its ripening phase, the skin of the berry then being easily peeled off, exposing the dorsal vascular bundles without damaging either these or the pulp tissue of the berry. The sites of sugar phloem unloading are thus made directly accessible and may be regulated by the buffer solution. In addition, the unloaded photoassimilates are easily transported into the buffer solution in the berry cup. With the berry-cup technique, it takes 60 min to purge the sugar already present in the apoplast, after which the amount of sugar in the buffer solution is a direct measure of the sugar unloading from the grape berry phloem. The optimum times for sampling were 20 or 30 min, depending on the type of experiment. Sugar phloem unloading was significantly inhibited by the inclusion of either 7.5 mm NaF or 2.5 mm PCMB in the buffer solution. This study indicates that sugar phloem unloading in ripening grape berries is via the apoplastic network and that the process requires the input of energy. The system was shown to be an appropriate experimental system with which to study sugar phloem unloading in ripening grape berries, and was applied successfully to the study of berry sugar unloaded from grapevines subjected to water stress. The results showed that water deficiency inhibits sugar unloading in grape berries. PMID- 12907467 TI - The role of roots and cotyledons as storage organs in early stages of establishment in Quercus crispula: a quantitative analysis of the nonstructural carbohydrate in cotyledons and roots. AB - Quercus seedlings have hypogeal cotyledons and tap roots, both of which act as storage organs. The importance of the storage function in the two organs may change as the seedling develops. Therefore, changes in carbohydrate reserves in cotyledons and roots of Q. crispula grown under 40 % and 3 % of full light from shoot emergence to the completion of the first leaf flush were monitored. In addition, a shoot-clipping treatment was performed to examine the relative contribution of the cotyledons and tap roots to resprouting. Cotyledons maintained large amounts of nonstructural carbohydrates during shoot development, and carbohydrates were still present in the cotyledons during the final phase of leaf flush. In addition, a notable increase in the amount of carbohydrates was observed in tap roots before leaf flush at both light levels. Since root development occurred before leaf flush, even in plants grown under 3 % light, the carbohydrate found in them presumably originated from seed reserves and was translocated to roots as storage reserves. When shoots were clipped at the leaf flushing stage, the amount of carbohydrate decreased only in the cotyledons after resprouting, suggesting that cotyledons act as the main storage organs during shoot development stages. However, it could be advantageous as a 'risk avoidance strategy' for the seedlings to store reserves in both cotyledons and roots, since cotyledons may be removed by predators during shoot development. PMID- 12907468 TI - Movement of water from old to young leaves in three species of succulents. AB - A hypothetical adaptive response of succulent plants to drought-stress is the redistribution of water from old to young leaves. We examined the effects of possible movement of water from old to young leaves in three succulent species, Carpobrotus edulis (weak CAM-inducible), Kalanchoe tubiflora (CAM) and Sedum spectabile (possibly a CAM-cycler or CAM-inducible). Old leaves were removed from plants, and photosynthesis, transpiration, f. wt : d. wt ratios, diurnal acid fluctuations, stomatal conductance and internal CO2 concentrations of the remaining young leaves were measured during drought-stress. Comparison was made with plants retaining old leaves. There was no evidence that water moved from old to young leaves during drought-stress as previously hypothesized. Only in drought stressed plants of K. tubiflora, were photosynthetic and transpiration rates of young leaves greater on shoots with old leaves removed compared with attached. There was a trend in all species for greater fluctuations in acidity in young leaves on shoots that lacked older leaves. For two of the three species studied, the f. wt : d. wt ratios of young leaves were greater under drought-stress, on shoots with old leaves removed than with them attached. Absence of old leaves may reduce competition for water with young leaves, which consequently have higher water content and greater photosynthetic rates. PMID- 12907469 TI - Suspected child abuse: the potential for justice to miscarry. PMID- 12907471 TI - Endocrine treatment of physiological gynaecomastia. PMID- 12907472 TI - General practitioners and occupational health professionals. PMID- 12907473 TI - Speak up! PMID- 12907475 TI - Judges rule that children can sue for wrongful diagnosis of abuse. PMID- 12907476 TI - South African government to withdraw antiretrovirals for pregnant mothers. PMID- 12907478 TI - Drug company employee who queried trial wins appeal. PMID- 12907479 TI - Baby milk company fined for advertising direct to consumers. PMID- 12907480 TI - Hong Kong under WHO spotlight after flu outbreak. PMID- 12907481 TI - "Yellow card" announcement used to head off concerns, experts claim. PMID- 12907482 TI - Medical prescription of heroin to treatment resistant heroin addicts: two randomised controlled trials. AB - OBJECTIVE: To determine whether supervised medical prescription of heroin can successfully treat addicts who do not sufficiently benefit from methadone maintenance treatment. DESIGN: Two open label randomised controlled trials. SETTING: Methadone maintenance programmes in six cities in the Netherlands. PARTICIPANTS: 549 heroin addicts. INTERVENTIONS: Inhalable heroin (n = 375) or injectable heroin (n = 174) prescribed over 12 months. Heroin (maximum 1000 mg per day) plus methadone (maximum 150 mg per day) compared with methadone alone (maximum 150 mg per day). Psychosocial treatment was offered throughout. MAIN OUTCOME MEASURES: Dichotomous, multidomain response index, including validated indicators of physical health, mental status, and social functioning. RESULTS: Adherence was excellent with 12 month outcome data available for 94% of the randomised participants. With intention to treat analysis, 12 month treatment with heroin plus methadone was significantly more effective than treatment with methadone alone in the trial of inhalable heroin (response rate 49.7% v 26.9%; difference 22.8%, 95% confidence interval 11.0% to 34.6%) and in the trial of injectable heroin (55.5% v 31.2%; difference 24.3%, 9.6% to 39.0%). Discontinuation of the coprescribed heroin resulted in a rapid deterioration in 82% (94/115) of those who responded to the coprescribed heroin. The incidence of serious adverse events was similar across treatment conditions. CONCLUSIONS: Supervised coprescription of heroin is feasible, more effective, and probably as safe as methadone alone in reducing the many physical, mental, and social problems of treatment resistant heroin addicts. PMID- 12907483 TI - Interpregnancy interval and risk of preterm birth and neonatal death: retrospective cohort study. AB - OBJECTIVE: To determine whether a short interval between pregnancies is an independent risk factor for adverse obstetric outcome. DESIGN: Retrospective cohort study. SETTING: Scotland. SUBJECTS: 89 143 women having second births in 1992-8 who conceived within five years of their first birth. MAIN OUTCOME MEASURES: Intrauterine growth restriction (birth weight less than the 5th centile for gestational age), extremely preterm birth (24-32 weeks), moderately preterm birth (33-36 weeks), and perinatal death. RESULTS: Women whose subsequent interpregnancy interval was less than six months were more likely than other women to have had a first birth complicated by intrauterine growth restriction (odds ratio 1.3, 95% confidence interval 1.1 to 1.5), extremely preterm birth (4.1, 3.2 to 5.3), moderately preterm birth (1.5, 1.3 to 1.7), or perinatal death (24.4, 18.9 to 31.5). They were also shorter, less likely to be married, and more likely to be aged less than 20 years at the time of the second birth, to smoke, and to live in an area of high socioeconomic deprivation. When the outcome of the second birth was analysed in relation to the preceding interpregnancy interval and the analysis confined to women whose first birth was a term live birth (n = 69 055), no significant association occurred (adjusted for age, marital status, height, socioeconomic deprivation, smoking, previous birth weight vigesimal, and previous caesarean delivery) between interpregnancy interval and intrauterine growth restriction or stillbirth. However, a short interpregnancy interval (< 6 months) was an independent risk factor for extremely preterm birth (adjusted odds ratio 2.2, 1.3 to 3.6), moderately preterm birth (1.6, 1.3 to 2.0), and neonatal death unrelated to congenital abnormality (3.6, 1.2 to 10.7). The adjusted attributable fractions for these associations were 6.1%, 3.9%, and 13.8%. The associations were very similar when the analysis was confined to married non smokers aged 25 and above. CONCLUSIONS: A short interpregnancy interval is an independent risk factor for preterm delivery and neonatal death in the second birth. PMID- 12907485 TI - Rate limiting factors in recruitment of patients to clinical trials in cancer research: descriptive study. PMID- 12907484 TI - Past exposure to sun, skin phenotype, and risk of multiple sclerosis: case control study. AB - OBJECTIVE: To examine whether past high sun exposure is associated with a reduced risk of multiple sclerosis. DESIGN: Population based case-control study. SETTING: Tasmania, latitudes 41-3 degrees S. PARTICIPANTS: 136 cases with multiple sclerosis and 272 controls randomly drawn from the community and matched on sex and year of birth. MAIN OUTCOME MEASURE: Multiple sclerosis defined by both clinical and magnetic resonance imaging criteria. RESULTS: Higher sun exposure when aged 6-15 years (average 2-3 hours or more a day in summer during weekends and holidays) was associated with a decreased risk of multiple sclerosis (adjusted odds ratio 0.31, 95% confidence interval 0.16 to 0.59). Higher exposure in winter seemed more important than higher exposure in summer. Greater actinic damage was also independently associated with a decreased risk of multiple sclerosis (0.32, 0.11 to 0.88 for grades 4-6 disease). A dose-response relation was observed between multiple sclerosis and decreasing sun exposure when aged 6 15 years and with actinic damage. CONCLUSION: Higher sun exposure during childhood and early adolescence is associated with a reduced risk of multiple sclerosis. Insufficient ultraviolet radiation may therefore influence the development of multiple sclerosis. PMID- 12907486 TI - Effect of national guidelines on prescription of methadone: analysis of NHS prescription data, England 1990-2001. PMID- 12907487 TI - Acute low back pain: systematic review of its prognosis. AB - OBJECTIVES: To describe the course of acute low back pain and sciatica and to identify clinically important prognostic factors for these conditions. DESIGN: Systematic review. DATA SOURCES: Searches of Medline, Embase, Cinahl, and Science Citation Index and iterative searches of bibliographies. MAIN OUTCOME MEASURES: Pain, disability, and return to work. RESULTS: 15 studies of variable methodological quality were included. Rapid improvements in pain (mean reduction 58% of initial scores), disability (58%), and return to work (82% of those initially off work) occurred in one month. Further improvement was apparent until about three months. Thereafter levels for pain, disability, and return to work remained almost constant. 73% of patients had at least one recurrence within 12 months. CONCLUSIONS: People with acute low back pain and associated disability usually improve rapidly within weeks. None the less, pain and disability are typically ongoing, and recurrences are common. PMID- 12907488 TI - Involving patients in medical education. PMID- 12907489 TI - Tennis elbow. PMID- 12907490 TI - Treatment of tennis elbow: the evidence. PMID- 12907491 TI - Neonatal alloimmune thrombocytopenia. PMID- 12907492 TI - Implantable devices for treating tachyarrhythmias. PMID- 12907493 TI - Establishing ethical trials for treatment and prevention of AIDS in developing countries. PMID- 12907494 TI - Ethics guidance for HIV prevention trials. PMID- 12907495 TI - No more free lunches: how to dance with porcupines. PMID- 12907496 TI - No more free lunches: doctors and journals in the South cut off drug companies at their peril. PMID- 12907497 TI - No more free lunches: it is time we all grew up. PMID- 12907498 TI - No more free lunches: relationship between industry and research does not bear scrutiny. PMID- 12907499 TI - No more free lunches: it is time to own up. PMID- 12907501 TI - No more free lunches: the market has us all in its grip. PMID- 12907502 TI - Dying for drugs: drug companies should not have to subsidise incompetent governments. PMID- 12907503 TI - Dying for drugs: drug companies should swear "Hippocratic" oath. PMID- 12907504 TI - Industry funding of patients' support groups: objectives of patients, clinicians, and industry are similar. PMID- 12907505 TI - Prognosis of entanglement could be serious. PMID- 12907506 TI - Industry funding of patients' support groups: declaration of competing interests is rare in Italian breast cancer associations. PMID- 12907507 TI - Interests of charities: diabetes UK has long recognised importance of transparency in funding. PMID- 12907508 TI - Interests of charities: arthritis care sets facts straight. PMID- 12907509 TI - On abandoning ties and avoiding nose rings: medical humour has implications for evidence based medicine... PMID- 12907510 TI - On abandoning ties and avoiding nose rings:. but ties may in truth pose danger to clinicians. PMID- 12907511 TI - On abandoning ties and avoiding nose rings: ... and economic analysis of tie wearing might be interesting. PMID- 12907512 TI - Telephone consultations are routinely used. PMID- 12907514 TI - Polio vaccine plans may run into problems in Nigeria. PMID- 12907515 TI - Australian ministers threaten restrictions on junk food advertising. PMID- 12907518 TI - Doctors in antarctica. PMID- 12907520 TI - Asthma and physical activity. PMID- 12907521 TI - Encouraging news from the antibiotic resistance front. PMID- 12907522 TI - Watch the p wave: it can change! PMID- 12907523 TI - Age and ARDS. PMID- 12907524 TI - Gut mucosal blood flow: regional regulation or systemic pressure dependence? PMID- 12907525 TI - Yet another look at noninvasive positive-pressure ventilation. PMID- 12907526 TI - Leisure-time physical activity patterns among US adults with asthma. AB - BACKGROUND: Little is known about the physical activity patterns among US adults who have asthma. METHODS: Using data for 165,123 respondents of the 2000 Behavioral Risk Factor Surveillance System, we examined leisure-time physical activity. RESULTS: After adjusting for age, about 30% of participants with current asthma (12,489 participants), 24% with former asthma (4,892 participants), and 27% who never had asthma (147,742 participants) were considered to be inactive (p < 0.001). After adjusting for age, the estimated energy expenditure from leisure-time physical activity was 206 kilocalories (kcal) per week lower among respondents with current asthma than among respondents with former asthma (p < 0.001) and 91 kcal/week lower than respondents who had never had asthma (p < 0.001). About 27% of participants with current asthma, 28% of participants with former asthma, and 28% of participants who had never had asthma were participating in recommended levels of physical activity. Walking was the most frequently reported activity for all three groups (respondents with current asthma, 39%; respondents with former asthma, 39%; and respondents who had never had asthma, 38%. Participants with asthma were less likely to engage in running (p < 0.001), basketball (p = 0.001), golf (p < 0.001), and weightlifting (p = 0.001) but were more likely to use an exercise bicycle (p = 0.035) than were participants without asthma. CONCLUSIONS: Like most US adults, the majority of those with asthma were not meeting the current recommendations for physical activity. PMID- 12907527 TI - Stabilization of an increasing trend in physician-diagnosed asthma prevalence in Saskatchewan, 1991 to 1998. AB - OBJECTIVES: To determine the prevalence of asthma, bronchitis, and COPD using the physician services database of the Saskatchewan Health Department from 1991 to 1998. DESIGN: Descriptive population-based study. SETTING: The Province of Saskatchewan, Canada. PARTICIPANTS: Residents of Saskatchewan covered by universal health care in the province. RESULTS: In all age groups, asthma prevalence increased between 1991 and 1995 and either was stable or declined between 1996 and 1998. Preschool children had the highest asthma prevalence during the study period, followed by children aged 5 to 14 years, young adults aged 15 to 34 years, and adults aged 35 to 64 years. Children aged 0 to 4 years and adults aged 35 to 64 years in the Registered Indian population had greater asthma prevalence than persons in other urban or rural populations during the study period. Asthma prevalence rates in rural populations were less than or similar to the rates of urban populations in all age groups during the study period. The prevalence of bronchitis was greater in the Registered Indian population than in urban and rural populations in all age groups throughout the study period. When persons who had visited a physician for bronchitis were excluded from the prevalence calculation, the original increases seen in asthma prevalence among very young children and older adults of Registered Indian origin disappeared, with the urban population having greater asthma prevalence in all age groups. In the Registered Indian population, adults aged 35 to 64 years had almost twofold increases in the prevalence of COPD in comparison to other Saskatchewan populations. CONCLUSIONS: Asthma prevalence, which had been on the increase in the 1980s and early 1990s, was either stable or declining during the latter part of 1990s in Saskatchewan. Preschool children and older adults from the Registered Indian population had greater asthma prevalence than did those from other Saskatchewan populations. Asthma prevalence among the rural populations was either similar or lower in comparison to the rates for the urban populations in all age groups during the study period. Further research is required to elucidate the findings in this study. PMID- 12907528 TI - The effect of smoking intervention and an inhaled bronchodilator on airways reactivity in COPD: the Lung Health Study. AB - BACKGROUND: The Lung Health Study (LHS), a 5-year, randomized, prospective clinical trial, studied the effects of smoking intervention and therapy with inhaled anticholinergic bronchodilators on FEV(1) in participants who were 35 to 60 years of age and had mild COPD. Participants were randomized into the following three groups: usual care; smoking cessation plus inhaled ipratropium bromide; and smoking cessation plus placebo inhaler. This report evaluates the effects of these interventions, demographic characteristics, smoking status, and FEV(1) changes on airway responsiveness (AR). METHODS AND RESULTS: Of 5,887 participants, 4,201 underwent methacholine challenge testing both at study entry and study completion. All groups increased AR during the 5-year period. The increase in AR was greatest in continuing smokers and was associated with a greater FEV(1) decline. An intent-to-treat analysis indicated no significant differences in AR changes among the three groups. CONCLUSIONS: Changes in AR over a 5-year period in the LHS were primarily related to changes in the FEV(1). The greater the decline in FEV(1), the greater the increase in AR. Smoking cessation had a small additional benefit in AR beyond its favorable effects on FEV(1) changes. PMID- 12907529 TI - Mortality and mortality-related factors after hospitalization for acute exacerbation of COPD. AB - BACKGROUND: Acute exacerbations form a major component of the socioeconomic burden of COPD. As yet, little information is available about the long-term outcome of patients who have been hospitalized with acute exacerbations, although high mortality rates have been reported. STUDY OBJECTIVE: The aim of this study was to investigate prospectively the outcome for all patients admitted to the hospital with acute exacerbations of COPD during hospital admission and after 1 year of follow-up. Furthermore, patient characteristics related to increased mortality rate were analyzed. DESIGN: We investigated prospectively the 1-year mortality rate and potential determinants of mortality for all patients admitted to the hospital with an acute exacerbation between January 1 and December 31, 1999. RESULTS: A total of 171 patients were included in the study. The mortality rate during hospital stay was 8%, increasing to 23% after 1 year of follow-up. Despite a comparable in-hospital mortality rate (6%), the 1-year mortality rate was significantly higher for patients admitted to the ICU for respiratory failure (35%). The multivariate Cox proportional hazards model was used to determine independent predictors of survival. Variables included in the regression model were age, sex, FEV(1), PaO(2), PaCO(2), body mass index, long-term use of oral corticosteroids, comorbidity index, and hospital readmissions. The maintenance use of oral glucocorticosteroids (relative risk [RR], 5.07; 95% confidence interval [CI], 2.03 to 12.64), PaCO(2) (RR, 1.17; 95% CI, 1.01 to 1.38), and age (RR, 1.07; 95% CI, 1.01 to 1.12) were independently related to mortality. CONCLUSION: We conclude that the prognosis for patients who have been admitted to the hospital for acute exacerbation of COPD is poor. Long-term use of oral corticosteroids, higher PaCO(2), and older age could be identified as risk factors associated with higher mortality. PMID- 12907530 TI - Specific expiratory muscle training in COPD. AB - BACKGROUND: There are several reports showing that expiratory muscle strength and endurance can be impaired in patients with COPD. This muscle weakness may have clinically relevant implications. Expiratory muscle training tended to improve cough and to reduce the sensation of respiratory effort during exercise in patients other than those with COPD. METHODS: Twenty-six patients with COPD (FEV(1) 38% predicted) were recruited for the study. The patients were randomized into two groups: group 1, 13 patients were assigned to receive specific expiratory muscle training (SEMT) daily, six times a week, each session consisting of 1/2 h of training, for 3 months; and group 2, 13 patients were assigned to be a control group and received training with very low load. Spirometry, respiratory muscle strength and endurance, 6-min walk test, Mahler baseline dyspnea index (before), and the transitional dyspnea index (after) were measured before and after training. RESULTS: The training-induced changes were significantly greater in the SEMT group than in the control group for the following variables: expiratory muscle strength (from 86 +/- 4.1 to 104 +/- 4.9 cm H(2)O, p < 0.005; mean difference from the control group, 24%; 95% confidence interval, 18 to 32%), expiratory muscle endurance (from 57 +/- 2.9% to 76 +/- 4.0%, p < 0.001; mean difference from the control group, 29%; 95% confidence interval, 21 to 39%), and in the distance walked in 6 min (from 262 +/- 38 to 312 +/- 47 m, p < 0.05; mean difference from the control group, 14%; 95% confidence interval, 9 to 20%). There was also a small but not significant increase (from 5.1 +/- 0.9 to 5.6 +/- 0.7, p = 0.14) in the dyspnea index. CONCLUSIONS: The expiratory muscles can be specifically trained with improvement of both strength and endurance in patients with COPD. This improvement is associated with increase in exercise performance and no significant change in the sensation of dyspnea in daily activities. PMID- 12907531 TI - The proportional Venn diagram of obstructive lung disease: two approximations from the United States and the United Kingdom. AB - STUDY OBJECTIVES: The nonproportional Venn diagram of obstructive lung disease (OLD) produced for the 1995 American Thoracic Society guidelines has not been quantified. We aim to quantify the proportion of the general population with OLD and the intersections of physician-diagnosed asthma, chronic bronchitis, and emphysema in the United States and the United Kingdom, and to examine the relationship to obstructive spirometry. DESIGN AND PARTICIPANTS: We analyzed data from the US National Health and Nutrition Examination (NHANES) III survey (1988 to 1994) and the UK General Practice Research Database for the year 1998. RESULTS: The areas of intersection among the three OLD conditions produced seven mutually exclusive disease groups. The asthma-only group was the largest proportion of OLD patients, accounting for 50.3% and 79.4% of all OLD patients in the United States and the United Kingdom, respectively, and decreased with increasing age. Overall, 17% and 19% of OLD patients in the United States and in the United Kingdom, respectively, reported more than one OLD condition, and this percentage increased with age. According to the spirometry data from NHANES III, only 37.4% of emphysema-only patients had objective airflow obstruction. The prevalence of airflow obstruction was significantly higher among participants with combinations of emphysema and chronic bronchitis (57.7%), with emphysema and asthma (51.9%), and with all three OLD diseases concomitantly (52.0%). CONCLUSION: Concomitant diagnosis of asthma, chronic bronchitis, or emphysema is common among OLD patients from the general population, particularly in adults aged > or = 50 years. PMID- 12907532 TI - A multicenter study of the effect of solution temperature on nasal potential difference measurements. AB - BACKGROUND: Nasal potential difference (PD) measurement quantifies the abnormal sodium and chloride transport that is characteristic of cystic fibrosis (CF) and has gained acceptance as both a diagnostic tool and outcome measure for new CF therapies. Because small changes in nasal PD-measured chloride transport are often an important component in evaluating new CF therapies, techniques to maximize sensitivity and reproducibility are essential. STUDY OBJECTIVE: To determine if administration of warmed nasal PD solutions (37 degrees C), instead of room temperature solutions (22 degrees C), results in significant increase in nasal PD-measured transepithelial chloride transport. DESIGN: Multicenter, prospective, cross-over trial of repeated measurements of nasal PD at 22 degrees C and 37 degrees C. RESULTS: Thirty-two healthy volunteers completed the study (four centers, each with 8 subjects). For 22 degrees C vs 37 degrees C, baseline (+/- SD) nasal PD (- 19.3 +/- 6.9 millivolts [mV] vs - 18.8 +/- 7.7 mV, p = 0.76), amiloride-sensitive PD (Delta PD, 10.4 +/- 5.6 mV vs 11.0 +/- 6.1 mV, p = 0.60), and low chloride response (Delta PD, - 10.0 +/- 8.0 mV vs - 8.0 +/- 7.1 mV, p = 0.13) were not statistically significantly affected by warming of solutions. Warming solutions to 37 degrees C dramatically increased the chloride transport response to isoproterenol (Delta PD, - 6.9 +/- 6.4 mV vs - 13.3 +/- 8.8 mV, p < 0.01) and the combined total response to low chloride and isoproterenol (Delta PD, - 16.9 +/- 9.5 mV vs - 21.3 +/- 11.9 mV, p = 0.01). The average increases observed with warming in isoproterenol and combined total responses were - 6.4 mV (95% confidence interval [CI], - 8.5 to - 4.3) and - 4.4 mV (95% CI, - 7.6 to - 1.1), respectively. CONCLUSIONS: Performing nasal PD studies with solutions at 37 degrees C instead of 22 degrees C increases the observed total chloride response by approximately 25% and the isoproterenol-dependent chloride response by approximately 95%. The Cystic Fibrosis Foundation Therapeutics Development Network now recommends warming of solutions as a standard procedure for nasal PD protocols. Utilization of warmed solutions will standardize nasal PD techniques across centers and potentially increase the ability to identify therapies that result in small incremental improvements in CF transmembrane conductance regulator function. PMID- 12907533 TI - Metabolic alkalosis contributes to acute hypercapnic respiratory failure in adult cystic fibrosis. AB - BACKGROUND: and study objectives: Patients with end-stage cystic fibrosis (CF) develop respiratory failure and hypercapnia. In contrast to COPD patients, altered electrolyte transport and malnutrition in CF patients may predispose them to metabolic alkalosis and, therefore, may contribute to hypercapnia. The aim of this study was to determine the prevalence of metabolic alkalosis in adults with hypercapnic respiratory failure in the setting of acute exacerbations of CF compared with COPD. DESIGN: Levels of arterial blood gases, plasma electrolytes, and serum albumin from 14 consecutive hypercapnic CF patients who had been admitted to the hospital with a respiratory exacerbation were compared with 49 consecutive hypercapnic patients with exacerbations of COPD. Hypercapnia was defined as a PaCO(2) of > or = 45 mm Hg. RESULTS: Despite similar PaCO(2) values, patients in the CF group were significantly more alkalotic than were those in the COPD group (mean [+/- SD] pH, 7.43 +/- 0.03 vs 7.37 +/- 0.05, respectively; p < 0.01). A mixed respiratory acidosis and metabolic alkalosis was evident in 71% of CF patients and 22% of COPD patients (p < 0.01). The mean concentrations of plasma chloride (95.1 +/- 4.9 vs 99.8 +/- 5.2 mmol/L, respectively; p < 0.01) and sodium (136.5 +/- 2.8 vs 140.4 +/- 4.5 mmol/L, respectively; p < 0.01) were significantly lower in the CF group, and the levels of serum albumin were significantly reduced (27.4 +/- 5.8 vs 33.7 +/- 4.8 mmol/L, respectively; p < 0.01). CONCLUSION: Metabolic alkalosis contributes to hypercapnic respiratory failure in adults with acute exacerbations of CF. This acid-base disturbance occurs in conjunction with reduced total body salt levels and hypoalbuminemia. PMID- 12907534 TI - Relation between lung function and RBC distribution width in a population-based study. AB - STUDY OBJECTIVES: Pulmonary function is dependent not only on smoking, but also on nutritional status. Since an increased RBC distribution width (RDW) has been associated with nutritional deficiencies, we postulated that RDW has an inverse relation to pulmonary function. The purpose of this study was to test this hypothesis. DESIGN AND SETTING: A cross-sectional study was conducted of a random sample of the general population in western New York. PARTICIPANTS: A total of 1,616 subjects of both genders who were aged 35 to 79 years and were free of respiratory disease. INTERVENTIONS: None. MEASUREMENTS: Pulmonary function was assessed from FEV(1), FVC, height, body weight, total pack-years of smoking, smoking status, hemoglobin concentration, and hematologic indexes, eosinophil count, education, and blood levels of retinol, beta-cryptoxanthin, and vitamin E. RESULTS: We found a direct relation between RDW and the number of pack-years of smoking and smoking status, and an inverse relation between FEV(1) and FVC with RDW, even when potentially confounding variables such as smoking were taken into account. When the variability of FEV(1) due to smoking was used for comparison, an additional 27% of that variability in FEV(1) was explained by variations in antioxidant vitamin levels, and another 24% by RDW. CONCLUSIONS: The results confirmed our hypothesis that there is an inverse relation between RDW and pulmonary function, and raise the possibility that RDW may be a biomarker for as yet unidentified nutrients that affect pulmonary function. PMID- 12907535 TI - Measurement characteristics of peak expiratory flow. AB - STUDY OBJECTIVES: To evaluate features of the peak expiratory flow (PEF) test protocol, and to characterize patterns of reproducibility in multiple PEF measurements. DESIGN: Cross-sectional study. SETTING: University population. PARTICIPANTS: Two hundred twenty-three healthy adults. INTERVENTIONS: Participants recorded five PEF measurements in each of five sessions per day for 1 week. MEASUREMENTS AND RESULTS: Patterns of within-session variability were characterized using a reproducibility criterion based on a large percentage difference between best trials and evidence of a maneuver-induced bronchospasm (MIB) indicated by successive drops of PEF values in a session. Although the maximum PEF value in a session occurred on the fourth or fifth trial 32% of the time, the change in PEF values was small. Supervision was associated with small improvements in level and reproducibility. Using a cutoff of 5% for defining reproducibility, 15% of all sessions were not reproducible. When averaged over each subject, 9% of the cohort had a mean difference > 5%. Overall, MIB was unusual and observed in 8% of all test sessions; however, MIB was more common among asthmatics and subjects with wheeze, atopy, or allergies than subjects without. By contrast, poor reproducibility was more common among smokers and subjects with cough and phlegm. CONCLUSIONS: These results illustrate that it may be unnecessary to supervise all sessions or collect more than three efforts. Results also suggest that reproducibility reflects smoking-related abnormalities, whereas MIB may reflect airways hyperresponsiveness. PMID- 12907536 TI - Peak physiologic responses to arm and leg ergometry in male and female patients with airflow obstruction. AB - STUDY OBJECTIVE: To investigate differences in work capacity for the arms and legs in patients with moderate-to-severe COPD. DESIGN: Cross-sectional investigation. PATIENTS: One hundred twenty-four patients (90 men and 34 women) aged 45 to 81 years with moderate-to-very severe COPD. FEV(1) ranged from 0.70 to 2.79 L/min (FVC, 1.73 to 5.77 L; FEV(1)/FVC, 24 to 69%). All patients were in stable condition at the time of testing and receiving a stable drug regime. MEASUREMENTS: Each patient completed a demographic and medical history questionnaire, pulmonary function studies (spirometry, lung volumes, and diffusion capacity), peak exercise ergometry with gas exchange for the arms and legs; they also rated their subjective assessment of perceived dyspnea and extremity fatigue using Borg scores during exercise. RESULTS: Patients were of comparable age, with men taller and heavier than women. Smoking history was significantly less for women (47.9 pack-years vs 66.6 pack-years for men) even though each group presented with equivalent age (p > 0.05). Women were less obstructed than men, with FEV(1)/FVC (mean +/- SD) of 46.5 +/- 10.9% vs 40.2 +/- 9.3%, respectively. Ventilatory limitation during exercise was noted for all patients studied. Peak work capacity was greater for men, and leg peak responses were greater than arm values for each gender. As airway obstruction increased, work capacity became more limited. Peak arm work achieved was 38.9 +/- 19.6 W, oxygen uptake (VO(2)) was 903.9 +/- 263.5 mL/min, and minute ventilation (VE) was 33.7 +/- 9.5 L. Peak leg work value was 62.9 +/- 24.8 W, VO(2) was 1,091.4 +/- 321.5 mL/min, and VE was 39.3 +/- 12.0 L. Hence, arm values were 62%, 83%, and 85% of the measured leg values, respectively. Dyspnea and extremity effort scores were similar for men and women, and for arms and legs. Regression analysis was used to derive prediction equations for arm work from measured leg ergometry testing. For watts of work, a three-variable model emerged explaining 66% of the variance; VO(2) yielded a four-variable model with 80% of the variance explained; and VE yielded a three-variable model explaining 72% of the variance. CONCLUSION: Arm work is reduced by 38% that of the legs, while more modest reductions are noted for VO(2) and VE, suggesting greater mechanical efficiency for leg work as compared to arm work. These data also suggest greater metabolic demand for respiratory muscles and arm ergometry. Dyspnea and extremity Borg scores were equivalent for each modality and level of airway obstruction studied, suggesting that perception plays an important role in limiting exercise, and that a threshold for termination of exercise may exist. Further, peak leg ergometry results can be used with pulmonary function indexes to predict peak arm workload in watts, VO(2), and VE. These data may be used to assist the clinician in prescribing rehabilitation or estimating arm exercise ability when arm testing is unavailable. PMID- 12907537 TI - Decreasing beta-lactam resistance in Pneumococci from the Memphis region: analysis of 2,152 isolates From 1996 to 2001. AB - STUDY OBJECTIVES: The Memphis region historically has had high pneumococcal antibiotic resistance rates. In recent years, we have seen a significant shift in antibiotic use away from beta-lactams toward the newer quinolones and macrolides. We hypothesized that these changes would cause a shift in pneumococcal antibiotic resistance patterns. DESIGN: Retrospective cohort study. SETTING: A large private hospital system. PATIENTS AND METHODS: We analyzed the antibiotic susceptibility patterns of 2,152 pneumococcal isolates obtained in the Memphis area from 1996 to 2001. Isolates were categorized as invasive or noninvasive and antibiotic resistance was classified according to the 2000 guidelines of the National Committee for Clinical Laboratory Standards. RESULTS: Over the study period, the proportion of penicillin-susceptible noninvasive pneumococcal isolates taken from children increased from 22 to 44% (p = 0.0004 [for trend across the 6-year period]). In noninvasive isolates taken from adults, penicillin susceptibility increased from 22 to 55% (p = 0.002), with a trend toward increasing sensitivity to cefotaxime (p = 0.02) in noninvasive isolates over the same period. The proportion of isolates with high-level penicillin resistance (ie, minimum inhibitory concentration, > or = 4 microg/mL) also decreased between 1996 and 2001 (p = 0.003). Clindamycin resistance in adult noninvasive isolates also declined (p = 0.002). The only adverse trend observed over this period was an increase in erythromycin resistance in noninvasive isolates from adults (p = 0.01). Resistance rates were significantly higher in children than in adults and were higher in noninvasive isolates than in invasive isolates. CONCLUSIONS: The stabilization of beta-lactam resistance rates in our region suggests that a continuous increase in pneumococcal resistance to antibiotics is not inevitable and may be avoidable. PMID- 12907539 TI - Measurements of HIV viral loads from different levels of the respiratory tract. AB - BACKGROUND: The lung is a common site of disease in HIV infection. Virus has been detected in BAL fluid (BALF) and saliva. However, the relationship between viral loads detected at different levels of the respiratory tract is unknown. METHOD: We measured simultaneous HIV viral loads in parotid saliva (PS), bronchial fluid (BF), BALF, and plasma by reverse transcription polymerase chain reaction in 20 HIV-infected individuals. RESULTS: HIV was detected in 53% of BALF samples, 15% of BF samples, 5% of PS samples, and 88% of plasma samples. Viral loads in plasma and BALF samples were positively correlated. There were significantly higher levels of HIV viral load in both plasma and BALF in subjects with CD4 counts of < 200 cells/ microL compared to those with higher counts. Antiretroviral therapy (ART) was associated with lower BALF and plasma viral loads, and the effect in BALF was independent of the plasma viral load. Interestingly, smoking also was associated with lower levels of both BAL and BF viral loads, independent of the plasma viral load. CONCLUSION: These data demonstrate that while HIV can be detected in the respiratory tract, the viral load is influenced by both local factors (ie, level of the respiratory tree and cigarette smoking) and systemic factors (ie, ART and peripheral CD4 count). PMID- 12907538 TI - An economic evaluation of sequential i.v./po moxifloxacin therapy compared to i.v./po co-amoxiclav with or without clarithromycin in the treatment of community acquired pneumonia. AB - STUDY OBJECTIVE: To evaluate costs, clinical consequences, and cost-effectiveness from a German and French health-care system perspective of sequential i.v./po moxifloxacin monotherapy compared to co-amoxiclav with or without clarithromycin (AMC +/- CLA) in patients with community-acquired pneumonia (CAP) who required parenteral treatment. METHODS: Costs and consequences over 21 days were evaluated based on clinical cure rates 5 to 7 days after treatment and health resource use reported for the TARGET multinational, prospective, randomized, open-label trial. This trial compared sequential i.v./po monotherapy with moxifloxacin (400 mg qd) to i.v./po co-amoxiclav (1.2 g i.v./625 mg po tid) with or without clarithromycin (500 mg bid) for 7 to 14 days in hospitalized patients with CAP. Since no country by-treatment interaction was found in spite of some country differences for length of hospital stays, resource data (antimicrobial treatment, hospitalization, and out-of-hospital care) from all centers were pooled and valued using German and French unit prices to estimate CAP-related cost to the German Sickness Funds and French public health-care sector, respectively. RESULTS: Compared to AMC +/- CLA, treatment with moxifloxacin resulted in 5.3% more patients achieving clinical cure 5 to 7 days after therapy (95% confidence interval [CI], 1.2 to 11.8%), increased speed of response (1 day sooner for median time to first return to apyrexia, p = 0.008), and a reduction in hospital stay by 0.81 days (95% CI, - 0.01 to 1.63) within the 21-day time frame. Treatment with moxifloxacin resulted in savings of 266 euro and 381 euro for Germany and France respectively, primarily due to the shorter length of hospital stay. Cost-effectiveness acceptability curves show moxifloxacin has a > or = 95% chance of being cost saving from French and German health-care perspectives, and higher probability of being cost-effective at acceptability thresholds up to 2,000 euro per additional patient cured. CONCLUSION: i.v./po monotherapy with moxifloxacin shows clinical benefits including increased speed of response and is cost-effective compared to i.v./po AMC +/- CLA in the treatment of CAP. PMID- 12907540 TI - Legionnaires disease and HIV infection. AB - STUDY OBJECTIVES: To compare the outcome of Legionnaires disease (LD) in patients with and without HIV infection. DESIGN: Retrospective review of clinical charts. SETTING: Six hundred-bed university hospital. PATIENTS: We studied the clinical findings of 64 patients without HIV and 15 patients with HIV. Patients with a serologic diagnosis only were not included. Patients with previous immunosuppressive therapy or transplant recipients were excluded from the former group. In the HIV group, the mean CD4 cell count was 347.5/ microL, plasma viral load was undetectable in 50% of the patients, and only one patient (7%) was receiving cotrimoxazole as prophylaxis against Pneumocystis carinii at the time of pneumonia. No differences were observed in the two groups with respect to community or nosocomial acquisition, delay in the initiation of appropriate treatment, the use of macrolides or fluoroquinolones, and Fine score in cases of community-acquired LD. RESULTS: Univariate analysis showed that time to apyrexia was longer, and respiratory symptoms, bilateral infiltrates in chest radiograph, hyponatremia, increase in aspartate aminotransferase and creatine phosphokinase (CK), and respiratory failure were more frequent in the HIV group. Mortality was greater in patients with HIV, achieving a statistically significant value of 20%; however, multivariate analysis only confirmed these differences with respect to the increase in CK. CONCLUSIONS: LD has a more severe clinical presentation and worse evolution in patients with HIV. PMID- 12907541 TI - Suboptimal erythropoietic response to hypoxemia in idiopathic pulmonary fibrosis. AB - BACKGROUND AND STUDY OBJECTIVES: Idiopathic pulmonary fibrosis (IPF) is a chronic inflammatory process characterized by severe derangement of gas exchange in the advanced stages of disease. However, erythrocytosis is infrequent in IPF. The aim of this study was to investigate the potential relation between the blunted erythropoietic response and the chronic inflammation. SUBJECTS: Nine patients (6 men and 3 women) with IPF and profound hypoxemia (PO(2) < 65 mm Hg) and 34 sex- and age-matched healthy volunteers participated in the study. METHODS: We evaluated the hematologic parameters, serum erythropoietin, tumor necrosis factor (TNF)-alpha, interleukin (IL)-6, and IL-8 levels. We also studied the development of burst-forming unit-erythroid (BFU-E)-derived colonies in semisolid methylcellulose cultures in blood samples from all patients. RESULTS: Hemoglobin and serum erythropoietin levels were almost comparable between the two studied groups. On the contrary, serum TNF-alpha, IL-6, and IL-8 values were significantly higher in patients with IPF (p < 0.05, p < 0.01, and p < 0.001, respectively). IPF sera induced a significant growth inhibition of erythroid bursts arising from mononuclear cells of either patients or control subjects compared with heat-inactivated AB serum (p < 0.05 and p < 0.01, respectively). Moreover, there was an apparent increment in the number of BFU-E colonies when patients' mononuclear cells were cultured in comparison with those of healthy subjects (p < 0.05). CONCLUSIONS: Our findings suggest that in IPF there is an increased number of primitive erythroid progenitors, which fail to proliferate and differentiate in vivo, suggesting a kind of ineffective erythropoiesis. As a consequence, hemoglobin levels do not rise in proportion to the severity of hypoxemia. Cytokines released from alveolar macrophages seem to have not only local but also systemic effects, since the serum of these patients directly suppressed erythropoiesis; however, the suboptimal erythropoietic response to hypoxia cannot be entirely attributed to this suppression. It is possible that several other factors interfere, synergistically or additively. PMID- 12907542 TI - Low hospital mortality in patients with acute interstitial pneumonia. AB - STUDY OBJECTIVES: To compare the presenting features and outcome of patients with acute interstitial pneumonia (AIP) with other patients with diffuse alveolar damage (DAD) and with historical control subjects. DESIGN: Retrospective chart review. SETTING: A large, urban, teaching hospital. INTERVENTIONS: Patients were classified into idiopathic (AIP group) and secondary causes of DAD (ARDS group) according to available clinical and microbiology data. AIP and ARDS cases were compared, and ARDS cases were analyzed for long-term outcome. MEASUREMENTS AND RESULTS: Twenty patients with pathologic diagnosis of DAD were identified. Four cases were excluded; eight cases of ARDS due to known etiologies were identified. These etiologies included pneumonia and sepsis (n = 6), cocaine use (n = 1), and carmustine chemotherapy (n = 1). Eight cases of AIP were found. When compared with the ARDS group, patients in the AIP group had a longer time from the onset of symptoms until hospital admission (16.8 +/- 15.7 days vs 2.2 +/- 1.0 days, p = 0.0015) and a shorter time from hospital admission to open-lung biopsy (8.3 +/- 3.0 days vs 15.5 +/- 9.5 days, p = 0.02) [mean +/- SD]. Seven of eight patients with AIP and four of eight patients with ARDS survived to hospital discharge (p = not significant). The 12.5% mortality rate for patients with AIP reported in this series was significantly lower than the previously reported cumulative rate of 69.5% (p = 0.0025). Follow-up in five AIP survivors for a mean of 7.6 +/- 3.5 years (range, 3 to 11 years) showed all to be without shortness of breath or relapse despite mild residual fibrosis on chest radiograph and mild-to-moderate restriction on pulmonary function tests (mean total lung capacity, 68.5 +/- 6.2% predicted). CONCLUSIONS: Our data support a favorable hospital and long-term outcome for patients with AIP, with no evidence of recurrence or progression to chronic interstitial lung disease. PMID- 12907543 TI - Acute right atrial strain: regression in normal as well as abnormal P-wave amplitudes with treatment of obstructive pulmonary disease. AB - OBJECTIVE: To evaluate right atrial (RA) "strain" as reflected by changes in P wave amplitude and vector in patients with COPD immediately before and immediately after beginning treatment of exacerbations. BACKGROUND: P-pulmonale (frequently temporary, reflecting acute RA strain) occurs under a variety of circumstances, including COPD. Emergency room (ED) ECGs in patients with acute exacerbations of COPD have suggested that P-pulmonale (P waves > or = 2.5 in leads II, III, and aVF) tends to resolve subsequent to acute treatment. RA strain is defined as a response to RA stress (probably transient pressure rise and/or acute RA enlargement) in patients with COPD. Since P-pulmonale occurs in a small minority of patients with COPD, we investigated dynamic changes in size and mean vector (axis) of all frontal plane P waves in the ED vs the immediate subsequent ward ECG in patients with acute exacerbations of COPD. METHODS: We prospectively compared P-wave amplitude in the ED with the first in-patient ECG in 50 consecutive patients with acute exacerbations of COPD and in 20 consecutive nonpulmonary control patients, analyzing only ECGs showing sinus rhythm and in which P waves were clearly recorded. Despite using a calibrated magnifying graticule, it was difficult to interpret a dynamic change if the initial ED ECG had P-wave amplitude < 1.5 mm in leads II and aVF. We selected lead II because it usually has the largest frontal plane P waves and also aVF to reflect the relative verticality of the mean P vector (axis). We performed a matched-pair analysis to compare the equality of means. RESULTS: Of the patients with COPD, only seven patients (14%) had classical P-pulmonale on the ED ECG. Forty-eight of 50 consecutive patients (96%) demonstrated a decrease in P-wave amplitude between ED and subsequent ward ECGs. Two patients showed no change. The mean differences of P-wave amplitude between ED and ward ECGs in lead II was 0.78 mm, and that in lead aVF was 0.8 mm. The difference of the mean P-axis between ED and ward ECGs was - 5.24 degrees (p < 0.0001 for all three measurements). There was no P-wave amplitude change in the control group between ED and ward ECGs. CONCLUSIONS: P wave amplitude in patients with COPD decreases once an acute exacerbation subsides. Thus, P-wave amplitude and vector are dynamic and could reflect reduced RA strain. We question the traditional (1935) absolute cutoff of 2.5 mm for P pulmonale as of limited value due to insensitivity, hence inappropriate for what this investigation demonstrates to be a continuous variable. PMID- 12907544 TI - Cardiac rupture complicating acute myocardial infarction in the direct percutaneous coronary intervention reperfusion era. AB - BACKGROUND: Cardiac rupture, an uncommon yet catastrophic complication after acute myocardial infarction (AMI), has been studied primarily in the prethrombolytic and thrombolytic therapy eras but not in the direct percutaneous coronary intervention (d-PCI) reperfusion therapy era. The aim of this study was to delineate the incidence, potential risks, timing of occurrence, clinical features, and outcomes of cardiac rupture complicating AMI after d-PCI. METHODS AND RESULTS: Between May 1993 and July 2002, a total of 1,250 patients with AMI underwent d-PCI in our hospital. Of these 1,250 patients studied, 12 patients (0.96%) had cardiac rupture (ventricular septal defect [VSD], three patients; left ventricular [LV] free wall rupture, nine patients] after d-PCI, with a mean (+/- SD) time of occurrence of 52.3 +/- 36.2 h. Three patients with VSD had an insidious presentation, and two of these patients (66.6%) survived after surgical intervention. However, nine patients with LV free wall rupture always presented with sudden and unanticipated hemodynamic collapse. Cardiopulmonary resuscitation was uniformly unsuccessful in patients with LV free wall rupture, and all patients died as a result of this complication within minutes of its onset. The 30-day mortality rate was significantly higher in patients with cardiac rupture than in patients without this complication (83.3% vs 8.2%, respectively; p < 0.001). Univariate analysis demonstrated that the left anterior descending artery was the most likely to be totally occluded in patients who had developed cardiac rupture (100% vs 66.4%, respectively; p = 0.033). Multiple stepwise logistic regression analysis demonstrated that the most significant factors associated with cardiac rupture were advanced age, female gender, and lower body mass index (BMI; all p < 0.05), whereas early reperfusion with d-PCI was an independent determinant of preventing this complication (p < 0.0001). CONCLUSION: Compared with the prethrombolytic era, our study showed that d-PCI had a favorable impact on reducing the incidence of cardiac rupture after AMI. Old age, female gender, lower BMI, and longer time to reperfusion carried a substantially increased risk of cardiac rupture after patients experienced AMIs. Early successful d-PCI was the most powerful determinant of the avoidance of this catastrophic complication after AMI. PMID- 12907545 TI - Oxygen cost of exercise is increased in heart failure after accounting for recovery costs. AB - STUDY OBJECTIVES: The oxygen cost during exercise has been reported to be decreased in patients with congestive heart failure (CHF), implying an increased efficiency (lower oxygen uptake [VO(2)] per Watt [VO(2)/W]); however, these studies ignored the oxygen debt that is increased in heart failure. SUBJECTS: The primary aim of this research was to evaluate the total oxygen cost (work VO(2)/W) during exercise and recovery in patients with heart failure as compared with healthy adults. DESIGN AND PATIENTS: We performed a retrospective analysis comparing the exercise VO(2)/W, the recovery VO(2)/W, the work VO(2)/W, and the VO(2)/W relationship above and below the ventilatory threshold (VT) in 11 healthy control subjects and 45 patients with CHF. RESULTS: The exercise VO(2)/W was decreased by 29% (p < 0.0001) in patients with CHF; however, the recovery VO(2)/W was increased by 167% (p < 0.0001) and the work VO(2)/W was increased by 14% in patients with CHF (p = 0.014). The VO(2)/W slope increased above the VT (+ 27%, p = 0.0017) in both normal subjects and patients with CHF, suggesting a decrease in efficiency above the VT. There was an inverse correlation (r = 0.646, p < 0.0001) between exercise VO(2)/W and recovery VO(2)/W, implying that subjects with a low exercise VO(2)/W were not efficient but rather accumulated a large oxygen debt that was repaid following completion of exercise. CONCLUSIONS: Heart failure is associated with lower exercise VO(2)/W; however, the patient with heart failure is not efficient, but rather accumulating a large oxygen debt (recovery VO(2)/W) that is repaid following exercise. In addition, the work VO(2)/W (including both exercise and recovery) is increased in patients with heart failure in comparison to control subjects, and correlates inversely with the percentage of predicted VO(2). The large recovery VO(2)/W is likely due to impaired oxygen delivery to exercising muscle during exercise. The increase in the work VO(2)/W is probably due to changes in skeletal muscle fiber type that occur in patients with heart failure (type I to type IIb). PMID- 12907546 TI - Prolonged oxygen uptake kinetics during low-intensity exercise are related to poor prognosis in patients with mild-to-moderate congestive heart failure. AB - STUDY OBJECTIVE: To investigate the prognostic value of oxygen uptake (O(2)) kinetics during low-intensity exercise in patients with congestive heart failure. DESIGN: Prospective cohort study. SETTING: Tertiary care center. PATIENTS: One hundred forty-six consecutive patients (128 men) with chronic heart failure, followed up for a mean (+/- SD) duration of 25 +/- 15 months. MEASUREMENTS: A treadmill exercise test was performed with "breath by breath" gas-exchange monitoring. VO(2) kinetics were defined as the VO(2) deficit (ie, Delta VO(2) x time[rest to steady state] - Sigma VO(2)[rest to steady state]) and mean response time (MRT) [ie, VO(2) deficit/Delta VO(2)]. Cardiac death, urgent cardiac transplantation, and hospitalization due to worsening heart failure were considered as the end points. RESULTS: Thirty patients (21%) died, 11 patients (8%) underwent urgent transplantation, and 32 patients (22%) were hospitalized. In univariate analysis, MRT was the most powerful predictor of survival, survival free of urgent transplantation, and survival free of hospitalization (hazard ratios [HRs] per 10 s, 1.65, 1.72, and 1.61, respectively; all p < 0.0001). The predictive value of MRT exceeded that of peak VO(2) (HR per mL/kg/min, 0.90; p = 0.02, 0.91; p = 0.007, and 0.95; p = 0.08, respectively). In multivariate analysis, MRT (HR per 10 s, 1.73; p = 0.0002), resting systolic BP (HR per 10 mm Hg, 0.65; p = 0.003), and the slope of the ventilatory response to exercise (HR per 10 U, 1.68; p = 0.02) were independent predictors of survival. CONCLUSIONS: Our results suggest that VO(2) kinetics are strongly related to outcome in heart failure patients. Since it has several additional advantages over peak exercise testing (eg, less time-consuming, less demanding for the patients, less dependent on motivation, and applicable in patients with limitations other than cardiopulmonary disease), it has the potential to become a prognostic test for the assessment of heart failure patients. PMID- 12907547 TI - Snoring in preschool children: prevalence and association with nocturnal cough and asthma. AB - INTRODUCTION: The association between snoring, nocturnal cough, and allergic symptoms in young children is not known. OBJECTIVE: To measure the prevalence of habitual snoring and its association with nocturnal cough, asthma, and hay fever in preschool children. SETTING: A cross-sectional study. SUBJECTS: Preschool children aged 2 to 5 years. METHOD: The data were collected in a cross-sectional study. A total of 974 children were randomly selected from two areas of Lismore and Wagga Wagga in New South Wales, Australia. RESULTS: The prevalence of snoring was 10.5%, with no gender difference (p = 0.99) or trend association with age (p = 0.58). The association between snoring and nocturnal cough was highly significant (odds ratio [OR], 3.68; 95% confidence interval [CI], 2.41 to 5.63; p = 0.001). This association was significant in both the nonasthmatic and asthmatic groups when examined separately. Snoring was also significantly associated with asthma (OR, 2.03; 95% CI, 1.34 to 3.10; p = 0.001). In subjects without hay fever, the association between snoring and asthma was also highly significant (41.2% vs 24.8%; OR, 2.12; 95% CI, 1.34 to 3.37; p = 0.001). CONCLUSION: The prevalence of snoring in preschool children was 10.5% for both genders. Snoring was significantly associated with both nocturnal cough and asthma. Because snoring, asthma, and nocturnal cough may have a common etiology, it is possible that effective treatment of one symptom may lead to reductions in the presence or severity of the other symptoms. PMID- 12907548 TI - Left ventricular hypertrophy is a common echocardiographic abnormality in severe obstructive sleep apnea and reverses with nasal continuous positive airway pressure. AB - STUDY OBJECTIVES: To determine cardiac structural abnormalities by echocardiography in subjects with severe obstructive sleep apnea (OSA), and to determine the long-term effects of nasal continuous positive airway pressure (CPAP) on such abnormalities. DESIGN: Polysomnography was conducted on oximetry screened patients who showed a desaturation index > 40/h and > or = 20% cumulative time spent below 90%. From these, 25 patients with severe OSA but without daytime hypoxemia underwent echocardiography prior to, then 1 month and 6 months following initiation of CPAP treatment. SETTING: Outpatient sleep disorders center. RESULTS: Of the 25 patients, 13 patients (52%) had hypertension by history or on physical examination. Baseline echocardiograms showed that severe OSA was associated with numerous cardiovascular abnormalities, including left ventricular hypertrophy (LVH) [88%], left atrial enlargement (LAE) [64%], right atrial enlargement (RAE) [48%], and right ventricular hypertrophy (16%). In all patients (intent to treat) as well as those patients compliant with CPAP therapy (84% > 3 h nightly), there was a significant reduction in LVH after 6 months of CPAP therapy as measured by interventricular septal distance (baseline diastolic mean, 13.0 mm; 6-month mean after CPAP, 12.3 mm; p < 0.02). RAE and LAE were unchanged after CPAP therapy. CONCLUSIONS: LVH was present in high frequency in subjects with severe OSA and regressed after 6 months of nasal CPAP therapy. PMID- 12907549 TI - Respiratory monitoring by means of an unattended device in children with suspected uncomplicated obstructive sleep apnea: a validation study. AB - STUDY OBJECTIVE: To compare an unattended device for cardiorespiratory monitoring (POLY-MESAM; MAP; Martinsried, Germany) [P-M] with classic nocturnal polysomnography (PSG) for diagnosis of obstructive sleep apnea (OSA) in children. DESIGN: Clinical setting. PATIENTS: Twelve children (age range, 3 to 6 years) with highly suspected uncomplicated OSA who underwent PSG and P-M on 2 consecutive laboratory nights in a balanced manner. MEASUREMENTS: Respiratory indexes were compared for P-M (automated analysis), hand-scored revised P-M (P Mrev), and PSG. Analysis of contingency for cutoff levels of respiratory disturbance index (RDI) of 5 and 10 and level of agreement between P-M, P-Mrev, and PSG by the concordance method were evaluated. RESULTS: Nine of twelve children (75%) had a PSG RDI > 5, while 41.7% had an RDI > 10, indicating moderate-to-severe OSA. P-M sensitivity (78%) increased with the increase of the RDI cutoff, and P-Mrev sensitivity reached 100% at the cutoff of 10. The specificity was low for RDI > 5 and increased only modestly at RDI > 10 (P-Mrev, 57%). Seven of 12 children (increasing to 9 children with P-Mrev) and 9 of 12 children (increasing to 11 children with P-Mrev) were correctly classified by the P-M unit when cutoffs of 5 and 10 were considered, respectively. As far as the agreement level is concerned, P-M underestimated the incidence of obstructive hypopnea and overestimated the number of central apnea cases. P-Mrev improved the latter measure. CONCLUSION: Based on these data, the P-M device cannot be advocated for common use in a clinical setting, but it may have a role in urgent screening for highly suspected moderate-to-severe OSA. PMID- 12907550 TI - Value of F-18 fluorodeoxyglucose positron emission tomography for predicting the clinical outcome of patients with aggressive lymphoma prior to and after autologous stem-cell transplantation. AB - STUDY OBJECTIVES: To determine and compare the values of positron emission tomography (PET) with F-18 fluorodeoxyglucose (FDG) and CT for predicting clinical outcome of patients with aggressive lymphoma undergoing salvage cytoreductive chemotherapy followed by high-dose chemotherapy and autologous stem cell transplantation (ASCT). PATIENTS AND METHODS: Forty-three patients with lymphoma who underwent ASCT with FDG-PET evaluation were studied. Group 1 (n = 20) patients (6 patients with Hodgkin disease [HD], and 14 patients with non Hodgkin lymphoma [NHL]) underwent PET 2 to 5 weeks after initiation of salvage chemotherapy, prior to ASCT. Group 2 (n = 23) patients (6 patients with HD, and 17 patients with NHL) underwent PET within a median interval of 2.4 months (range, 2 to 6 months) after ASCT. MEASUREMENTS AND RESULTS: Study end points were complete remission, relapse, or death. In group 1, 8 of 20 patients (40%) were disease free after a median follow-up of 13.3 months; 12 patients relapsed or died. PET findings were true-negative in 7 of 8 patients and true-positive in 11 of 12 patients who relapsed after ASCT. In group 2, 9 of 23 patients (39%) were disease free after a median follow-up of 16.5-months; 14 patients relapsed. PET findings were true-negative in 8 of 9 patients and true-positive in 13 of 14 patients who relapsed. Positive and negative predictive values of PET were 92% and 88% (group 1) and 93% and 89% (group 2), respectively. Predictive accuracy values of PET were 90% and 91% for group 1 and group 2, respectively, vs 58% and 67% for CT (p < 0.05). CONCLUSIONS: PET findings but not CT results were strongly correlated with disease-free survival (p < 0.01). Our results show that FDG-PET can be used to predict the post-ASCT outcome of lymphoma patients with high accuracy. PMID- 12907551 TI - The cost-effectiveness of low-dose CT screening for lung cancer: preliminary results of baseline screening. AB - BACKGROUND: Low-dose CT scan screening greatly improves the likelihood of detecting small nodules and, thus, of detecting lung cancer at a potentially more curable stage. METHODS: To evaluate the cost-effectiveness of a single baseline low-dose CT scan for lung cancer screening in high-risk individuals, data from the Early Lung Cancer Action Project (ELCAP) was incorporated into a decision analysis model comparing low-dose CT scan screening of high-risk individuals (ie, those > or = 60 years with at least 10 pack-years of cigarette smoking and no other malignancies) to observation without screening. Cost-effectiveness was expressed as the incremental cost per year of life saved. The analysis adopted the perspectives of the health-care system. The probability of the different outcomes following the decision either to screen or not to screen an individual at risk was based on data from ELCAP and the Surveillance, Epidemiology, and End Results Registry or published data, respectively. The cost of the screening and treatment of patients with lung cancer was established based on data from the New York Presbyterian Hospital's financial system. The base-case analysis was conducted under the assumption of similar aggressiveness of screen-detected and incidentally discovered lung cancers and then was followed by multiple sensitivity analyses to relax these assumptions. RESULTS: The incremental cost effectiveness ratio of a single baseline low-dose CT scan was 2,500 US dollars per year of life saved. The base-case analysis showed that screening would be expected to increase survival by 0.1 year at an incremental cost of approximately 230 US dollars. Only when the likelihood of overdiagnosis was > 50% did the cost effectiveness ratio exceed 50,000 US dollars per year of life saved. The cost effectiveness ratios were also relatively insensitive to estimates of the potential lead-time bias. CONCLUSIONS: A baseline low-dose CT scan for lung cancer screening is potentially highly cost-effective and compares favorably to the cost-effectiveness ratios of other screening programs. PMID- 12907552 TI - Clinical equivalence of two cytokeratin markers in mon-small cell lung cancer: a study of tissue polypeptide antigen and cytokeratin 19 fragments. AB - STUDY OBJECTIVES: We have longstanding experience with tissue polypeptide antigen (TPA), a tumor marker of the cytokeratin (CK) family. In the mid-1990s, a new CK marker, CK 19 fragments (CYFRA 21-1), became popular and widely accepted. This is the first study specifically designed to compare the two markers. DESIGN: Analysis of a single institution database over a 3-year period (ie, 1998 to 2000). SETTING: Community-based hospital and second referral level institution for a province of 500,000 people. PATIENTS: The study included 180 new consecutive patients (143 men) with pathologically documented non-small cell lung cancer (NSCLC), who were observed during and after treatment, and eventually were assessed for status. INTERVENTIONS: Anthropometric, clinical, and laboratory data, including TPA and CYFRA 21-1 serum levels, were recorded prospectively. Standard nonparametric tests, Kaplan-Meyer survival analyses, Cox proportional hazards models, receiver-operating characteristic (ROC) curves, and estimates were used for statistical analysis. MEASUREMENTS AND RESULTS: A total of 1,299 twin TPA and CYFRA 21-1 serum assays (180 performed at diagnosis and 1,119 performed during or after treatment) were obtained. Intermarker correlation tests revealed incredibly high Spearman rho indexes, ranging from 0.935 at diagnosis to 0.813 to 0.921 at the different follow-up times. The substantial equivalence of the two tests explained all the other results, as follows: their similar profile of correlation with the other variables (objective treatment response: TPA rho, 0.456; CYFRA 21-1 rho, 0.463; follow-up performance status: rho range, 0.424 to 0.435); their superimposable capability to predict important clinical situations (eg, recognizing a metastatic disease at diagnosis with areas under the ROC curve of 0.742 and 0.706, respectively); their nearly identical prognostic significance (the D statistic of the goodness-of-fit of a multivariate survival model: TPA, 851.0; CYFRA 21-1, 851.6). CONCLUSIONS: In most of their traditional clinical applications the two serum tests are equivalent because of their virtual identity. We strongly recommend using a CK test in the evaluation of each NSCLC patient. The choice between TPA and CYFRA 21-1 can be based on nonclinical factors, such as the laboratory experience or preference, and the cost of the two kits. PMID- 12907553 TI - Nitric oxide metabolites are not reduced in exhaled breath condensate of patients with primary ciliary dyskinesia. AB - STUDY OBJECTIVES: To investigate whether nitric oxide (NO) metabolites would be reduced in children affected by primary ciliary dyskinesia (PCD). DESIGN: Single center observational study. PATIENTS: Fifteen children with PCD (seven boys; mean [+/- SEM] age, 10.3 +/- 0.7 years; mean FEV(1), 73 +/- 2.1% predicted) were recruited along with 14 healthy age-matched subjects (seven boys; mean age, 11.5 +/- 0.4 years; mean FEV(1), 103 +/- 5% predicted). INTERVENTIONS: We assessed the levels of nitrite (NO(2)(-)), NO(2)(-)/NO(3)(-) (NO(2)(-)/NO(3)(-)), and S nitrosothiol in exhaled breath condensate, exhaled NO, and nasal NO from children with PCD compared to those in healthy children. MEASUREMENTS AND RESULTS: The mean exhaled and nasal NO levels were markedly decreased in children with PCD compared to those without PCD (3.2 +/- 0.2 vs 8.5 +/- 0.9 parts per billion [ppb], respectively [p < 0.0001]; 59.6 +/- 12.2 vs 505.5 +/- 66.8 ppb, respectively [p < 0.001]). Despite the lower levels of exhaled NO in children with PCD, no differences were found in the mean levels of NO(2)(-) (2.9 +/- 0.4 vs 3.5 +/- 0.3 microM, respectively), NO(2)(-)/NO(3)(-) (35.2 +/- 5.0 vs 34.3 +/- 4.5 microM, respectively), or S-nitrosothiol (1.0 +/- 0.2 vs 0.6 +/- 0.1 microM, respectively) between children with PCD and healthy subjects. CONCLUSION: These findings suggest that NO synthase activity may not be decreased as much as might be expected on the basis of low exhaled and nasal NO levels. PMID- 12907554 TI - Exhaled nitric oxide as a predictor of exercise-induced bronchoconstriction. AB - INTRODUCTION: Exercise-induced bronchoconstriction (EIB) is present in 40 to 90% of patients with asthma. Exhaled NO (eNO) levels have been correlated with bronchial hyperresponsiveness to methacholine, and have correlated with the degree of decrease in FEV(1) with exercise. The purpose of our study was to examine whether eNO measurements prior to or after exercise could be used as a surrogate marker of exertional bronchoconstriction in a population referred specifically for the evaluation of EIB. METHODS: We studied 50 consecutive subjects without a history of asthma who were referred for the clinical evaluation of EIB. eNO levels were measured prior to exercise challenge and every 5 min for a total of 30 min after exercise. Forced expiratory flows were measured prior to and serially after exercise challenge. RESULTS: Seven subjects had a decrease in FEV(1) of > or = 15% with exercise. The mean eNO level prior to exercise was 41 parts per billion (ppb) [median +/- SD, 23 +/- 42.2 ppb] in the EIB group and 25.6 ppb (median, 19.95 +/- 18.47 ppb) in the group without EIB. A receiver operator characteristic curve yielded a value of 0.636. When using an eNO level of < 12 ppb, the sensitivity, specificity, negative predictive value, and positive predictive value for EIB were 1.0, 0.31, 0.19, and 1.0, respectively; therefore, no one with a baseline eNO of < 12 ppb demonstrated EIB. CONCLUSIONS: No subjects with very low pre-exercise eNO levels (< 12 ppb) demonstrated bronchial hyperresponsiveness to exercise. eNO measurement may obviate the need for bronchoprovocation testing in patients who complain of exertional dyspnea. PMID- 12907555 TI - Deactivation of alveolar macrophages in septic neutropenic ARDS. AB - STUDY OBJECTIVES: Neutrophils often have been involved in the pathophysiology of ARDS. However, authentic ARDS has been described in patients with severe neutropenia, suggesting the presence of other potential mechanisms that are responsible of this syndrome. Alveolar macrophages (AMs) could be involved in the development of ARDS, and so we decided to study AM activation in neutropenic patients. PATIENTS: We designed a prospective study and enrolled two subgroups of consecutive patients (group A, 18 patients; group B, 22 patients) with septic ARDS. In the first period, 7 of 18 patients were neutropenic, and in the second period 10 of 22 patients were neutropenic. All neutropenic patients were treated with granulocyte colony-stimulating factor (G-CSF). MEASUREMENTS AND RESULTS: In group A, BAL fluid samples were analyzed for differential and total cell counts, and alveolar activation marker expression (ie, human leukocyte antigen [HLA]-DR locus) was determined. Basal and lipopolysaccharide (LPS)-stimulated production of tumor necrosis factor, interleukin (IL)-1 beta, IL-6, and IL-10 was evaluated in group B. In neutropenic patients, the BAL fluid total cell count and the neutrophil absolute count was significantly lower compared to those in nonneutropenic patients (p = 0.029 and p = 0.046, respectively). HLA-DR expression on AMs was significantly decreased (p = 0.016), and the percentage of AMs expressing HLA-DR was also significantly lower (p = 0.041). In neutropenic patients, the mean percentage of AMs expressing HLA-DR was significantly lower in deceased patients compared to survivors (30 +/- 7 vs 43 +/- 1, respectively; p = 0.047). Basal AMs released cytokines was comparable between the two groups; however, LPS stimulation yielded a deactivation of AMs in neutropenic patients. CONCLUSION: These results suggest a deactivation and/or hypoactivation of AMs in septic ARDS patients. This deactivation/hypoactivation could be linked to the use of G-CSF as this molecule has been shown to generate a down-regulation of HLA-DR expression. PMID- 12907556 TI - Effect of age on the development of ARDS in trauma patients. AB - STUDY OBJECTIVES: The purpose of this study was to determine the independent effect of age on the risk of developing ARDS in patients with trauma. DESIGN: Prospective cohort study. SETTING: Level I trauma center. MEASUREMENTS AND RESULTS: A total of 4,020 consecutive trauma patients who were > 12 years of age were identified through the Harborview Medical Center Trauma Registry over a 3 year period. During this time, 484 of the trauma patients (12%) developed ARDS, as identified by the Harborview Medical Center ARDS Registry. Patients who developed ARDS were, on average, older (mean [+/- SD] age, 44.0 +/- 18.8 vs 40.2 +/- 20.0 years, respectively; p < 0.0001) and had higher injury severity scores (23.7 +/- 11.3 vs 18.0 +/- 10.3, respectively; p < 0.0001) than trauma patients who did not develop ARDS. The maximum unadjusted odds ratio for developing ARDS was 2.93 (95% confidence interval, 1.91 to 4.50) for the group 60 to 69 years of age compared to the group 13 to 19 years of age. Patients aged > or = 80 years had an equal risk of developing ARDS compared to those age 13 to 19 years. CONCLUSIONS: Age demonstrated a complex relationship with risk for ARDS development. Older patients showed increasingly higher risks for ARDS development up to 60 to 69 years of age, when the risk for ARDS declined. We concluded that older patients are at significantly greater risk of developing ARDS when compared to younger patients, while the oldest patients may be at less risk. PMID- 12907557 TI - Diffuse alveolar hemorrhage following allogeneic bone marrow transplantation in children. AB - BACKGROUND: Diffuse alveolar hemorrhage (DAH) is a frequent life-threatening complication of bone marrow transplantation (BMT) in adults. This noninfectious pulmonary disorder is rarely reported following BMT in neonates and children. STUDY OBJECTIVES: To review the clinical features and course of children who underwent allogeneic BMT and developed DAH in the posttransplant period. DESIGN: A retrospective 6-year chart review. SETTING: Pediatric ICU in a university hospital. PATIENTS AND INTERVENTIONS: At total of 138 children who had undergone allogeneic BMT for nonmalignant (n = 66) or malignant (n = 72) diseases. MEASUREMENTS AND RESULTS: Six of 138 children (4.3%) aged 2 months to 10 years (male/female ratio, 1:1) developed DAH. Each had a fulminant course with rapidly developing severe respiratory failure, mandating mechanical ventilation within 24 h following symptom onset. They were all treated with methylprednisolone, 6 mg/kg/d for 3 days. Only one child survived, and there have been no sequelae at 2 years post-BMT. Four children died of respiratory causes, and one died of multiorgan failure. CONCLUSIONS: DAH is a potentially fatal respiratory complication that should be included early in the differential diagnosis of acute respiratory failure in children following allogenic BMT for both malignant and nonmalignant diseases. Therapy with high doses of steroids apparently do not affect the course of the disease. PMID- 12907558 TI - Simplified method to measure respiratory-related changes in arterial pulse pressure in patients receiving mechanical ventilation. AB - STUDY OBJECTIVES: Measuring respiratory-related changes in arterial pulse pressure is useful to guide fluid expansion in hemodynamically compromised patients. In the absence of automation, this can be uneasy in clinical practice. The objective of this study was to test an alternative approach (expiratory pause) that should be easier to apply. DESIGN: Prospective observational study comparing two measurement methods of a biological variable. PATIENTS: Seventeen patients receiving mechanical ventilation without spontaneous respiratory activity, with an arterial indwelling catheter, exhibiting respiratory-related fluctuations in arterial pressure. SETTING: Ten-bed respiratory ICU in a 2,000 bed university hospital. INTERVENTION: Analysis of clinically gathered data without specific experimental intervention. MEASUREMENTS: Determinations of the change in arterial pulse pressure observed during ventilatory cycling (Delta Pp,dyn) ["dynamic"] and change in arterial pulse pressure observed during expiratory pauses (Delta Pp,stat) ["static"] were performed to assess respiratory mechanics, and comparison of the two sets of data (correlation, Bland and Altman, Passing and Bablok regression). RESULTS: Delta Pp,dyn and Delta Pp,stat were strongly correlated (R = 0.964; 95% confidence interval, 0.917 to 0.987; p < 0.0001), with a good level of agreement (mean difference, 0.016; lower limit of agreement, - 0.087; upper limit, 0.120) and no systematic difference. CONCLUSION: Measuring respiratory-related Delta Pp,stat provides data that seem interchangeable with Delta Pp,dyn, providing an easy means to routinely obtain this information. PMID- 12907559 TI - Development of respiratory syncytial virus "bronchiolitis" in guinea pigs does not reflect an allergic predisposition in the host. AB - BACKGROUND: Respiratory syncytial virus (RSV) infection causes bronchiolitis in a minority of children. Using a guinea pig model to determine if an allergic predisposition in the host increases permissiveness to RSV infection or severity of experimental "bronchiolitis," we compared the effects of RSV inoculation between strain 2 (allergy-resistant) and strain 13 (allergy-susceptible) inbred animals. METHODS: One month-old, juvenile guinea pigs were classified into four groups (eight guinea pigs per group): (group 1) strain 2, uninfected; (group 2) strain 13, uninfected; (group 3) strain 2, RSV infected; and (group 4) strain 13, RSV infected. Seven days after inoculation, the animals were studied by the following: viral plaque assays for quantification of intrapulmonary RSV; immunohistochemical localization of RSV antigens in lung tissue sections; physiologic assessment of airway obstruction and nonspecific bronchial hyperresponsiveness; quantitative histology of airway T lymphocytes, neutrophils, and eosinophils; and semiquantitative reverse transcriptase-polymerase chain reaction for levels of messenger RNA expression of a panel of proinflammatory cytokines and chemokines. RESULTS: Significantly higher titers of replicating RSV were isolated from the lungs of strain 13 vs strain 2 animals (p < or = 0.001). The two guinea pig strains showed similar cell types with positive viral immunostaining; RSV-associated changes in airway obstruction and nonspecific bronchial hyperresponsiveness; airway T cells, neutrophils, and eosinophils; and messenger RNA expression of cytokines and chemokines. CONCLUSIONS: Strain 13 guinea pigs show increased pulmonary RSV replication than strain 2 animals, but this increased permissiveness to the virus is not reflected by more severe virus induced changes in airway obstruction, nonspecific bronchial hyperresponsiveness, airway inflammation, or gene expression of proinflammatory cytokines and chemokines. PMID- 12907560 TI - Defensive role of pleural mesothelial cell sialomucins in tumor metastasis. AB - STUDY OBJECTIVES: Sialomucin complex (SMC) is a heterodimeric glycoprotein, and is found on the surfaces of the mesothelia of the pleura, pericardium, and peritoneum. Sialomucins play a significant role in adhesion as well as in defense. In this study, we hypothesized that pleural mesothelial cells (PMCs) express SMC and thus prevent the adherence of ovarian cancer cells (HTB-77) to the pleura. METHODS: PMCs were plated, and the adherence of HTB-77 cells was observed using a cytofluor. The PMC monolayer was pretreated with sialidase, and HTB-77 adherence was observed using cytofluor. In another set of HTB-77 cells, adherence was observed when the PMC monolayer was pretreated with supernatants of HTB-77 cells. Last, supernatants of HTB-77 cells were assayed for sialidase activity. RESULTS: The removal of SMC by sialidase greatly increased the adherence of HTB-77 cells to the PMC monolayer, which was statistically significant (p < 0.05). Similar results were obtained when the PMC monolayer was pretreated with the supernatants of HTB-77 cells. Supernatants of HTB-77 cells showed the presence of sialidase. CONCLUSIONS: The presence of SMC on the PMC acts as a defense layer, and its removal by sialidase increases the susceptibility of the PMC layer to the adherence of malignant cells and to increased metastasis. HTB-77 cells also express sialidase, which by its action on the monolayer aids in the adherence of tumor cells to the pleural surface. PMID- 12907561 TI - Effects of dopexamine and positive end-expiratory pressure on intestinal blood flow and oxygenation: the perfusion pressure perspective. AB - OBJECTIVE: To evaluate the net effects of the concomitant use of positive end expiratory pressure (PEEP) and dopexamine on intestinal tissue perfusion and oxygenation during predefined artificial reductions in intestinal perfusion pressure (IPP). DESIGN: Prospective, self-controlled, experimental study. SETTING: University hospital research laboratory. SUBJECTS: Seven female pigs. MEASUREMENTS: In barbiturate-anesthetized pigs, we measured mesenteric blood flow (QMES) [by transit-time ultrasonic flowmetry], jejunal mucosal perfusion (by laser Doppler flowmetry), and tissue PO(2) (by microoximetry). Based on blood sampling, we calculated the intestinal net lactate production and oxygenation. INTERVENTIONS: These measurements and calculations were performed at three predefined and controlled IPP levels, which were obtained by an adjustable clamp around the superior mesenteric artery. At each IPP level, measurements were performed prior to and during PEEP (10 cm H(2)O), both with and without simultaneous dopexamine infusions (at 0.5 and 1.0 microg/kg/min). RESULTS: Within the IPP range of 77 to 33 mm Hg, intestinal perfusion and oxygenation were maintained irrespective of whether PEEP and/or dopexamine were applied or not. At IPP < 33 mm Hg, QMES and intestinal oxygenation deteriorated, resulting in regional net lactate production. At this IPP range, tissue oxygen perfusion was entirely pressure-dependent, and even small reductions in IPP led to prominent increases in intestinal net lactate production. Dopexamine did not modify this pattern. CONCLUSIONS: We describe maintained intestinal tissue oxygen perfusion within a wide perfusion pressure range. Within this perfusion pressure range, PEEP did not induce any adverse regional circulatory effects. Below the perfusion pressure range for effective autoregulation, intestinal tissue oxygen perfusion deteriorated, and regional ischemia occurred. In this situation, dopexamine was unable to counteract IPP-dependent decreases in intestinal tissue oxygen perfusion. The regional ischemic threshold can be defined either as an IPP of < 33 mm Hg or as an intestinal tissue PO(2) of < 45 mm Hg. PMID- 12907562 TI - Acute applications of noninvasive positive pressure ventilation. AB - Noninvasive positive-pressure ventilation (NPPV) has been used increasingly to treat acute respiratory failure (ARF). The best indications for its use are ARF in patients with COPD exacerbations, acute pulmonary edema, and immunocompromised states. For these indications, multiple controlled trials have demonstrated that therapy with NPPV avoids intubation and, in the case of COPD and immunocompromised patients, reduces mortality as well. NPPV is used to treat patients with numerous other forms of ARF, but the evidence is not as strong for its use in those cases, and patients must be selected carefully. The best candidates for NPPV are able to protect their airway, are cooperative, and are otherwise medically stable. Success is optimized when a skilled team applies a well-fitted, comfortable interface. Ventilator settings should be adjusted to reduce respiratory distress while avoiding excessive discomfort, patient ventilator synchrony should be optimized, and adequate oxygenation should be assured. The appropriate application of NPPV in the acute care setting should lead to improved patient outcomes and more efficient resource utilization. PMID- 12907563 TI - Endovascular stent grafting of descending thoracic aortic aneurysms. AB - The treatment of descending thoracic aortic aneurysms using endovascular stents is one of the more recent advances in treatment and is receiving increasing attention as it is a less invasive alternative to open surgical repair. Although the technology is still primitive, significant improvements have lately been made in the design and deployment of the endovascular stent-grafts. Aortic stent grafts were used initially to exclude abdominal, and later thoracic, aortic true and false aneurysms. These prostheses have been increasingly used to treat aneurysms, dissections, and traumatic ruptures of the descending thoracic aorta with good early and mid-term outcomes. Although the long-term outcome of patients with aneurysms of the descending thoracic aorta after stent graft implantation has not been investigated, continued refinement of the endovascular approaches has decreased the need for conventional open thoracic aortic aneurysm repair, especially in patients who are at a high risk for standard surgery because of advanced age or the presence of comorbid diseases. The placement of endoluminal stent-grafts to exclude the dissected or ruptured site of thoracic aortic aneurysms is a technically feasible and relatively safe procedure. With the rapid development of endovascular approaches, the treatment of the descending thoracic aortic aneurysms might alter even more, but an extended follow-up is necessary to determine the longer term outcome. Historical perspectives, advantages, device considerations, complications, and current perspectives of the endovascular stent grafting of the descending thoracic aortic aneurysms are elaborated on. PMID- 12907564 TI - Technical considerations related to the minute ventilation/carbon dioxide output slope in patients with heart failure. AB - BACKGROUND: The minute ventilation (VE)-carbon dioxide output (VCO(2)) relationship has recently been demonstrated to have prognostic significance in the heart failure (HF) population. However, the method by which the VE/VCO(2) slope is expressed has been inconsistent. METHODS: One hundred eighty-eight subjects, who had received diagnoses of HF, underwent exercise testing. Two VE/VCO(2) slope calculations were made, one using exercise data prior to the ventilatory threshold (VT), and one using all data points from rest to peak exercise. Four separate peak exercise VE/VCO(2) slope calculations also were derived with unaveraged, 10-s, 30-s, and 60-s ventilatory expired gas sampling intervals. RESULTS: Although univariate Cox regression analysis demonstrated pre VT and peak VE/VCO(2) slope calculations to both be significant predictors of cardiac-related mortality and hospitalization (p < 0.001), the peak classification scheme was significantly better (p < 0.01). The ventilatory expired gas-sampling interval that was used did not impact the predictive ability of the peak VE/VCO(2) slope. CONCLUSION: Although both the pre-VT and peak VE/VCO(2) slope calculations were prognostically significant, the peak expression was superior. The sampling interval did not appear to have a significant impact on prognostic utility. We hope that the results of the present study will contribute to the standardization of the VE/VCO(2) slope and will enhance its clinical application. PMID- 12907565 TI - Evaluation and management of a patient with congestive heart failure. PMID- 12907566 TI - A patient with anemia and a paraspinal chest mass. PMID- 12907567 TI - Use of an ultrathin bronchoscope in the assessment of central airway obstruction. AB - STUDY OBJECTIVE: To assess the utility of an ultrathin bronchoscope (UB) in the assessment of central airway obstruction (CAO). DESIGN: Prospective evaluation SETTING: Tygerberg Hospital, a tertiary teaching hospital. PATIENTS: Consecutive patients referred to the Lung Unit with CAO. INTERVENTIONS: Fiberoptic bronchoscopy (FOB) was performed with a prototype UB (Olympus BF XP40; Olympus Europe; Hamburg, Germany; outer diameter, 2.8 mm; working channel, 1.2 mm). The UB was used whenever a standard bronchoscope (SB) could not pass the obstruction or could not be tolerated by the patient. MEASUREMENTS AND RESULTS: Data relating to indication and performance of FOB, patient demographics, utility in establishing a diagnosis, and planning definitive management were documented. Twenty-four patients (17 men; mean age, 46 years) were studied. Twelve patients (50%) had malignant CAO, 8 patients (33%) had benign tracheal stenosis, 3 patients (12.5%) had stent occlusion, and 1 patient (4%) had bilateral vocal cord paralysis. In 42% of patients, an initial attempt at passing the obstruction with an SB had failed. Vocal cords or trachea were involved in 62% of patients. The mean luminal occlusion was 84% of the total airway lumen (range, 50 to 100%). One complication (desaturation) led to early termination of FOB. In all but three patients with complete obstruction, the UB was able to pass the CAO and allowed assessment of the obstruction and the distal airways (87%). CONCLUSION: UB-FOB was useful and safe in the assessment of patients with CAO from both benign and malignant disease. It aided in establishing a diagnosis and/or planning of definitive management in all patients examined. PMID- 12907568 TI - Statins, inflammation, and sepsis: hypothesis. AB - Sepsis and septic shock are complex inflammatory syndromes. Multiple cellular activation processes are involved, and many humoral cascades are triggered. Statins have anti-inflammatory properties. Our preliminary observations indicate that patients receiving therapy with statins may have a lower incidence of severe sepsis. We hypothesize that statins have a strong protective effect against sepsis by virtue of diverse anti-inflammatory effects that are independent of their lipid-lowering ability. PMID- 12907569 TI - Diastolic heart failure demystified. AB - The mystery of diastolic heart failure (DHF), described by authorities as a "puzzle" and a "clinical paradox," stems from the following misperception: (1) that the normal ejection fraction implies normal cardiac output (CO), (2) that therefore low CO is not operative (it is rarely mentioned in relation to the pathophysiology of DHF), and (3) the congestive phenomena are due to the stiff left ventricle. In fact, a normal ejection fraction is not a reliable indicator of normal CO; low CO is the fundamental pathophysiologic abnormality of all heart failure (HF), whether systolic and/or diastolic (or, indeed, "high output"); and increased ventricular stiffness is not the principal cause of congestion in DHF. Pathophysiologic explorations supporting these understandings further reveal the following: (1) the premise that a clinical event as dramatic as acute pulmonary edema (systolic and/or diastolic) would be contingent on similarly dramatic acute hypertensive or ischemic ventricular dysfunction, while intuitive, is unsubstantiated, and there is an alternate explanation satisfying both theoretical and clinical observations; (2) contrary to general perception, DHF is no more vulnerable to diuretic-induced hypotension than systolic HF; (3) heart rate reduction should not yet be considered an established therapeutic goal in DHF; (4) since HF is HF whether systolic and/or diastolic, studies are likely to show that therapeutic similarities outweigh differences except as the various agents might modify the underlying structural and/or functional pathology; (5) although long evident that HF occurs by only two mechanisms (systolic dysfunction and/or diastolic dysfunction), it has only recently been acknowledged that the mere exclusion of one is diagnostic of the other; and (6) the definition of HF currently in widespread use is unnecessarily confounded by neglect of the fundamental distinction between ventricular dysfunction and failure. PMID- 12907570 TI - Acute respiratory failure with thrombocytopenia in a 47-year-old woman after hiking in the Sierras. PMID- 12907571 TI - Massive pulmonary pseudotumor. AB - The term pulmonary pseudotumor may be used to describe a well-demarcated interlobar pleural effusion. Pseudotumors are located within pulmonary fissures and are commonly associated with congestive heart failure and other processes that cause transudative pleural effusions. Pseudotumors are typically diagnosed presumptively on chest radiographs based on their lenticular configuration. We report a case of a massive pseudotumor opacifying one third of the right hemithorax on a frontal radiograph. CT scan of the chest showed a loculated effusion within the oblique fissure measuring 10 x 5 cm. The Hounsfield unit characteristic of the effusion was similar to that of freely layering liquid in the contralateral hemithorax that was shown to be a transudate. The pseudotumor resolved with medical management over 8 weeks. This case proves that even a massive pulmonary pseudotumor will resolve with conservative management. PMID- 12907572 TI - Blue rubber bleb nevus syndrome: endobronchial involvement presenting as chronic cough. AB - This case report describes a rare presentation of chronic cough secondary to endobronchial involvement with blue rubber bleb nevus syndrome (BRBNS) lesions. BRNBS is a rare syndrome characterized with cavernous hemangiomas involving the skin and GI tract. We report the case of a 37-year-old woman, with known BRBNS, who acquired an intractable cough with a radiograph revealing multiple pulmonary nodules. A bronchoscopy demonstrated various bluish, raised, and hypervascular lesions characteristic for BRBNS involving the right mainstem bronchus and segmental bronchi. To our knowledge, this is the first report of endobronchial involvement with BRBNS. PMID- 12907573 TI - Lymphangioleiomyomatosis and SZ alpha(1)-antitrypsin disease: a unique combination? AB - We describe a case of a 37-year-old female ex-smoker with lymphangioleiomyomatosis and SZ alpha(1)-antitrypsin disease who underwent successful bilateral sequential lung transplantation. While this disease combination has not been described previously, we recommend vigilance for the possibility of such combinations in patients with chronic lung disease. The possible mechanisms of interaction resulting from both disease processes are discussed. PMID- 12907574 TI - Catamenial pneumothorax: can all cases be explained by the pore hypothesis? PMID- 12907575 TI - Gastroesophageal reflux common in patients with sleep apnea rather than snorers without sleep apnea. PMID- 12907576 TI - Success rates for nortriptyline. PMID- 12907577 TI - Family-witnessed resuscitation. PMID- 12907578 TI - Clarifying cardiac decortication procedure. PMID- 12907579 TI - Another view of end-of-life care. PMID- 12907580 TI - Increasing statistical accuracy. PMID- 12907583 TI - Oral irritation by mustard oil: self-desensitization and cross-desensitization with capsaicin. AB - We investigated the temporal pattern of oral irritation elicited by sequential application of mustard oil (allyl-isothiocyanate), and whether it exhibits self desensitization and cross-desensitization with capsaicin. Mustard oil (0.125%, 40 micro l) was sequentially applied to one side of the tongue at 1 min intervals, and subjects rated the intensity of the irritant sensation elicited by each stimulus. Ratings successively declined across trials, indicating desensitization. In contrast, sequential application of capsaicin (10 ppm) elicited irritation that increased in intensity across trials (sensitization). To test for self-desensitization by mustard oil, a 10 min hiatus was imposed following the series of unilateral mustard oil stimuli, after which mustard oil was applied to both sides of the tongue. In a two-alternative forced-choice paradigm, subjects chose which side had stronger irritation and also independently rated the irritant intensity on each side. A significant majority of subjects chose the side not previously receiving mustard oil as more intense, and assigned significantly higher intensity ratings to that side, indicating self desensitization. In two additional sessions, the same paradigm was used to show mustard oil cross-desensitization of irritation elicited by capsaicin, and capsaicin cross-desensitization of irritation from mustard oil. In a final session, sequential application of mustard oil at faster (20 s) intervals initially evoked a sensitizing pattern followed by desensitization. The temporal patterns of oral irritation exhibited by mustard oil, and its reciprocal cross desensitization with capsaicin, are similar to those of menthol and nicotine. PMID- 12907584 TI - Quantification of chemical vapors in chemosensory research. AB - Studies of olfaction and chemesthesis often rely on nominal, liquid-phase dilutions to quantify the chemicals tested, even though the associated vapor concentrations constitute the actual stimuli. For more than a decade now, our systematic studies of the olfactory and chemesthetic potency of members of homologous chemical series have routinely included quantification of vapors via gas chromatography. This article depicts the relationships between liquid- and vapor-phase concentrations for 60 volatile organic compounds and summarizes the theoretical and technical factors influencing these relationships. The data presented will allow other investigators working with these materials to express them as vapor concentrations even when they lack the resources to perform the analytical measurements. The paper represents a step toward creation of a practical archive for vapor quantification in chemosensory science. PMID- 12907585 TI - Structure-activity studies with pheromone-binding proteins of the gypsy moth, Lymantria dispar. AB - Pheromone olfaction in the gypsy moth, Lymantria dispar, involves accurate distinction of compounds with similar structure and polarity. The identified sex pheromone is (7R,8S)-2-methyl-7,8-epoxyoctadecane, 1a, and a known antagonist is (7Z)-2-methyloctadec-7-ene, 4a. The first step in pheromone olfaction is binding of odorants by small, soluble pheromone-binding proteins (PBPs), found in the pheromone-sensing hairs. We have studied the molecular determinants recognized by the two PBPs found in the gypsy moth, using three pheromone/PBP binding assays. Results indicate that (i) PBPs bind analogs of the pheromone with some discrimination; (ii) PBPs experience enhancement of binding when presented with 1a or its enantiomer and 4a simultaneously; and (iii) the binding enhancement is also seen at high ligand:PBP ratios. We found no evidence of allostery, so the synergistic binding effects and the concentration effect may only be explained by multimerization of PBPs with each other, which leads to more than one population of binding sites. We suggest that the enhanced ligand binding at high ligand:PBP ratios may serve to sequester excess ligand and thereby attenuate very strong signals. PMID- 12907586 TI - Monkey electrophysiological and human psychophysical responses to mutants of the sweet protein brazzein: delineating brazzein sweetness. AB - Responses to brazzein, 25 brazzein mutants and two forms of monellin were studied in two types of experiments: electrophysiological recordings from chorda tympani S fibers of the rhesus monkey, Macaca mulatta, and psychophysical experiments. We found that different mutations at position 29 (changing Asp29 to Ala, Lys or Asn) made the molecule significantly sweeter than brazzein, while mutations at positions 30 or 33 (Lys30Asp or Arg33Ala) removed all sweetness. The same pattern occurred again at the beta-turn region, where Glu41Lys gave the highest sweetness score among the mutants tested, whereas a mutation two residues distant (Arg43Ala) abolished the sweetness. The effects of charge and side chain size were examined at two locations, namely positions 29 and 36. The findings indicate that charge is important for eliciting sweetness, whereas the length of the side chain plays a lesser role. We also found that the N- and C-termini are important for the sweetness of brazzein. The close correlation (r = 0.78) between the results of the above two methods corroborates our hypothesis that S fibers convey sweet taste in primates. PMID- 12907587 TI - High-resolution analysis of the spatio-temporal activity patterns in rat olfactory bulb evoked by enantiomer odors. AB - Understanding how mammals process olfactory stimuli has motivated the development of tools and techniques which permit the simultaneous study of finely structured spatial and temporal patterns of neural activity. A technique is described that uses an array of 32 penetrating microelectrodes implanted bilaterally into the dorsal aspect of rat olfactory bulb to investigate the responses of mitral and tufted neurons to stimulation with simple enantiomer odor pairs at a number of concentrations. It is shown that stable, simultaneous recordings from up to 49 single- and multi-units can be performed for periods of up to 14 h. We show that such odors evoke unique spatial and fast-temporal activity patterns which may subserve odor discrimination. This technique is extensible to other systems neuroscience investigations of olfactory sensory processing. PMID- 12907588 TI - Extracellular transduction events under pulsed stimulation in moth olfactory sensilla. AB - In natural conditions, pheromones released continuously by female moths are broken in discontinuous clumps and filaments. These discontinuities are perceived by flying male moths as periodic variations in the concentration of the stimulus, which have been shown to be essential for location of females. We study analytically and numerically the evolution in time of the activated pheromone receptor (signaling) complex in response to periodic pulses of pheromone. The 13 reaction model considered takes into account the transport of pheromone molecules by pheromone binding proteins (PBP), their enzymatic deactivation in the perireceptor space and their interaction with receptors at the dendritic membrane of neurons in Antheraea polyphemus sensitive to the main pheromone component. The time-averaged and periodic properties of the temporal evolution of the signaling complex are presented, in both transient and steady states. The same time averaged response is shown to result from many different pulse trains and to depend hyperbolically on the time-averaged pheromone concentration in air. The dependency of the amplitude of the oscillations of the signaling complex on pulse characteristics, especially frequency, suggests that the model can account for the ability of the studied type of neuron to resolve repetitive pulses up to 2 Hz, as experimentally observed. Modifications of the model for resolving pulses up to 10 Hz, as found in other neuron types sensitive to the minor pheromone components, are discussed. PMID- 12907589 TI - Influences of feedback and ascending and descending trial presentations on perithreshold odor detection performance. AB - The influences of feedback and ascending and descending trial sequences on the ability of 135 college-aged subjects to detect phenyl ethyl alcohol odorant concentrations ranging from 10(-9) to 10(-5.5) v/v were examined in a two alternative forced-choice test paradigm. At the highest concentrations, ascending trial sequences produced better performance than descending trial sequences; the reverse was true at the lowest concentrations. There was a tendency for feedback to improve performance marginally at the lowest two odorant concentrations presented. In the region associated with a traditional detection threshold calculation (i.e. at the 75% performance point in a two-choice detection task), no influences of feedback or direction of trial sequence were apparent. These data indicate that the effects of explicit feedback and trial sequence direction depend upon the segment of the peri-threshold stimulus concentration continuum evaluated. PMID- 12907590 TI - Understanding creaminess perception of dairy products using free-choice profiling and genetic responsivity to 6-n-propylthiouracil. AB - Flavor and texture contribute to the perception of creaminess in dairy products, but the nature of this interaction is not well understood. Previous studies on the genetic ability to perceive the bitter compound 6-n-propylthiouracil (PROP) reveal the existence of individual differences in creaminess perception. The objective of the present study was to use PROP-classified subjects to gain insight into this individual variation to better understand the cues for creaminess in dairy products, and to ascertain the contributions of flavor and texture to the integrated perception of creaminess. Ten nontasters and 10 supertasters of PROP participated in the study. Subjects evaluated nine commercial dairy products using Free-Choice Profiling (FCP), a type of descriptive analysis that allows subjects to rate products on individual lists of descriptors. Generalized Procrustes Analysis was used to develop separate consensus spaces for nontasters and supertasters. The models for both groups accounted for approximately 54% of the variance in the data and were resolved in two dimensions (a dairy flavor/texture axis and a sweet-sour continuum). The products were arranged in a similar pattern along the dimensions in both models. However, nontasters used a limited number of simple terms (sour, sweet, milky and mouthcoating) to describe the products, whereas supertasters used a more complex vocabulary (rich, buttery, creamy, light, grainy, gritty and sandy). The model for nontasters gave equal weight to the sweet-sour and dairy flavor/texture dimensions (28 and 26% variance, respectively); whereas, the model for supertasters relied more heavily on the dairy flavor/texture dimension (34% variance), and less so on the sweet-sour dimension (20% variance). These data suggest that the overall impression of creaminess was similar for nontasters and supertasters, but the cues the two groups used to judge creaminess differed. PMID- 12907591 TI - Evidence for deficiencies in perceptual and semantic olfactory processes in Parkinson's disease. AB - Olfactory deficits have been reported in Parkinson's disease (PD) and are thought to represent a sensitive marker of the disease. The aim of the present study was to examine the differential contribution in olfactory dysfunction of perceptual and semantic processes of odours in PD patients. Twenty-four PD patients (12 males and 12 females) and 24 control subjects (12 males and 12 females) were tested. The experiment included two sessions. Initially, 12 odorants were delivered, one per minute. For each odour, subjects were asked to rate intensity, pleasantness, familiarity and edibility using linear rating scales. The odorants were again presented and the subjects were asked to identify them. The four olfactory judgements and odour identification were severely disturbed in PD patients when compared to control subjects. These findings demonstrate major deficits for all cognitive tasks of olfactory judgement in PD, and suggest that PD is associated with disruption of olfactory areas situated in the temporal lobes and also in the prefrontal cortex. PMID- 12907592 TI - Parasitic wasps learn and report diverse chemicals with unique conditionable behaviors. AB - Parasitoids exploit numerous chemical cues to locate hosts and food. Whether they detect and learn chemicals foreign to their natural history has not been explored. We show that the parasitoid Microplitis croceipes can associate, with food or hosts, widely different chemicals outside their natural foraging encounters. When learned chemicals are subsequently detected, this parasitoid manifests distinct behaviors characteristic with expectations of food or host, commensurate with prior training. This flexibility of parasitoids to rapidly link diverse chemicals to resource needs and subsequently report them with recognizable behaviors offers new insights into their foraging adaptability, and provides a model for further dissection of olfactory learning related processes. PMID- 12907593 TI - The antioxidant conundrum in cancer. AB - The health-related effects of interactions between reactive oxygen species (ROS) and dietary antioxidants and the consequences of dietary antioxidant supplementation on human health are by no means clear. Although ROS, normal byproducts of aerobic metabolism, are essential for various defense mechanisms in most cells, they can also cause oxidative damage to DNA, proteins, and lipids, resulting in enhanced disease risk. Dietary antioxidants (e.g., vitamin E, vitamin C, beta-carotene, and selenium), as well as endogenous antioxidant mechanisms, can help maintain an appropriate balance between the desirable and undesirable cellular effects of ROS. However, any health-related effects of interactions between dietary antioxidants and ROS likely depend on the health status of an individual and may also be influenced by genetic susceptibilities. Clinical studies of antioxidant supplementation and changes in either oxidative status, disease risk, or disease outcome have been carried out in healthy individuals, populations at risk for certain diseases, and patients undergoing disease therapy. The use of antioxidants during cancer therapy is currently a topic of heated debate because of an overall lack of clear research findings. Some data suggest antioxidants can ameliorate toxic side effects of therapy without affecting treatment efficacy, whereas other data suggest antioxidants interfere with radiotherapy or chemotherapy. Overall, examination of the evidence related to potential interactions between ROS and dietary antioxidants and effects on human health indicates that consuming dietary antioxidant supplements has pros and cons for any population and raises numerous questions, issues, and challenges that make this topic a fertile field for future research. Overall, current knowledge makes it premature to generalize and make specific recommendations about antioxidant usage for those at high risk for cancer or undergoing treatment. PMID- 12907594 TI - PMEPA1, an androgen-regulated NEDD4-binding protein, exhibits cell growth inhibitory function and decreased expression during prostate cancer progression. AB - PMEPA1 was originally identified as a highly androgen-induced gene by serial analysis of gene expression in androgen-treated LNCaP prostate cancer (CaP) cells. PMEPA1 expression is prostate abundant and restricted to prostatic epithelial cells. PMEPA1-encoded protein shows high sequence homology to a mouse N4wbp4-encoded protein that binds to Nedd4 protein, an E3 ubiquitin-protein ligase involved in ubiquitin-dependent, proteasome-mediated protein degradation. Studies from our and other laboratories have suggested the role of PMEPA1 in cell growth regulation as noted by androgen induction of PMEPA1 expression, elevated PMEPA1 expression in nontumorigenic revertants of tumor cell lines after chromosome 8p transfer, and PMEPA1 expression alterations (up- or down regulation) in human tumors. Here, we demonstrate that PMEPA1 protein through its PY motifs interacts with WW domains of the human NEDD4 protein. Exogenous expression of PMEPA1, in widely used CaP cell lines DU145, PC3, LNCaP, and LNCaP sublines (C4, C4-2, and C4-2B), conferred cell growth inhibition, and at least one of the PY motifs of PMEPA1 may be involved in its cell growth inhibitory functions. Quantitative expression analysis of PMEPA1 in paired normal and tumor cells of 62 patients with primary CaP revealed tumor cells associated decreased expression in 40 of 62 patients that were significantly associated with higher pathologic stage and serum prostate-specific antigen. Taken together, PMEPA1 negatively regulates growth of androgen responsive or refractory CaP cells, and these functions may be mediated through the interaction of PMEPA1 with the NEDD4 protein involved in the ubiquitin-proteasome pathway. Loss or reduced PMEPA1 expression in CaP further suggests for its role in prostate tumorigenesis. PMID- 12907595 TI - Complementary effects of platelet-derived growth factor autocrine stimulation and p53 or Ink4a-Arf deletion in a mouse glioma model. AB - INK4a-ARF and p53 inactivation are common but rarely concurrent findings in glioblastoma multiforme. Here we demonstrate that experimental deletion of either tumor suppressor gene cooperates with retrovirally expressed platelet-derived growth factor (PDGF)-B regarding both tumor latency and frequency in a mouse brain tumor model. We find indications of PTEN down-regulation and increased Akt phosphorylation in both types of null tumors (although more prominent in p53-/- tumors) suggesting a possible mechanism for this synergism. This is the first time that the cooperative tumorigenic effects of PDGF-B stimulation and p53 loss of function are demonstrated in an in vivo model, establishing a functional link between two common molecular changes of human secondary glioblastoma multiforme. PMID- 12907596 TI - PML is required for homeodomain-interacting protein kinase 2 (HIPK2)-mediated p53 phosphorylation and cell cycle arrest but is dispensable for the formation of HIPK domains. AB - Here we demonstrate that endogenous human homeodomain-interacting protein kinase (HIPK) 2 and the highly homologous kinase HIPK3 are found in a novel subnuclear domain, the HIPK domains. These are distinct from other subnuclear structures such as Cajal bodies and nucleoli and show only a partial colocalization with promyelocytic leukemia (PML) nuclear bodies (PML-NBs). A kinase inactive HIPK2 point mutant is localized in the nucleoplasm. The occurrence of HIPK domains in PML-/- fibroblasts reveals their independence from the PML protein. HIPK2 can be almost completely recruited to PML-NBs by the PML isoform PML IV, but not by PML III. PML IV-mediated recruitment of HIPK2 does not rely on its kinase function and also occurs in PML-/- fibroblasts, showing that this PML isoform is sufficient for recruitment of HIPK2. Whereas the architecture of HIPK domains is PML independent, HIPK2-mediated enhancement of p53-dependent transcription, p53 serine 46 phosphorylation and the antiproliferative function of HIPK2 strictly rely on the presence of PML. PMID- 12907597 TI - Insulin-like growth factor binding protein 2 enhances glioblastoma invasion by activating invasion-enhancing genes. AB - Comparison of gene expressing profiles between gliomas with different grades revealed frequent overexpression of insulin-like growth factor binding protein 2 (IGFBP2) in glioblastoma (GBM), the most advanced stage of glioma. To determine whether IGFBP2 is involved in the proliferative and invasive nature of GBM, we established stable SNB19 GBM cell lines that overexpress IGFBP2. Although there was no marked difference in the cell growth between IGFBP2 overexpressing SNB19(BP2) lines when compared with the control cells, these clones showed significantly increased invasive rates when compared with the parental or vector transfected SNB19 cells. Total RNAs from controls and SNB19(BP2) clones were used for microarray analysis to detect IGFBP2-mediated alterations in gene expression. When compared with parental or vector-transfected control cells, SNB19(BP2) cells consistently showed 3-5-fold increase in the expression of matrix metalloproteinase-2 (MMP-2) as well as other invasion related genes. Increased MMP-2 expression in SNB19(BP2) cells was subsequently confirmed by real time reverse-transcription PCR, Western blotting, and gelatin zymography. Furthermore, consistent with increased MMP-2 expression in SNB19(BP2) cells, transient transfection of a MMP-2 promoter/luciferase reporter also resulted in 3-6-fold higher luciferase activity in SNB19(BP2) cells than in parental or vector transfected control cells. Finally, tissue microarray analysis of 68 GBM tissue specimens showed a significant correlation between the overexpression of IGFBP2 and elevated MMP-2 expression. Taken together, our data provide evidence that IGFBP2 contributes to glioma progression in part by enhancing MMP-2 gene transcription and in turn tumor cell invasion. PMID- 12907599 TI - Functional FAS promoter polymorphisms are associated with increased risk of acute myeloid leukemia. AB - The FAS (TNFRSF6/CD95/APO-1) gene is silenced in many tumor types, resulting in an inability to respond to proapoptotic signals. The FAS promoter is polymorphic, including a G to A substitution at -1377 bp and an A to G substitution at -670 bp, which occur within SP1 and signal transducers and activators of transcription 1 transcription factor binding sites, respectively. In a case-control study of adult acute myeloid leukemia (AML), we show a significantly increased risk of AML associated with heterozygotes (GA) and homozygote variants (AA) at position -1377 bp (32.3% in cases versus 22.0% in controls; odds ratio, 1.69; 95% confidence interval, 1.32-2.16). Extended haplotype analysis revealed that the -1377A/-670A haplotype was significantly associated with disease (3% versus 0.5%; odds ratio, 6.72; 95% confidence interval, 3.13-14.51). These data suggest that variation in the FAS gene promoter may affect FAS gene expression and modulate apoptotic signaling, contributing to an increased risk of AML. PMID- 12907598 TI - p27(Kip1) induces quiescence and growth factor insensitivity in tamoxifen-treated breast cancer cells. AB - Tamoxifen, a selective estrogen-receptor modulator, is effective in the treatment and prevention of breast cancer, but therapeutic resistance is common. Pure steroidal antiestrogens are efficacious in tamoxifen-resistant disease and, unlike tamoxifen, arrest cells in a state of quiescence from which they cannot reenter the cell cycle after growth factor stimulation. We now show that in hydroxytamoxifen-treated cells, transduction of the cell cycle inhibitor p27(Kip1) induces quiescence and insensitivity to growth stimulation by insulin/insulin-like growth factor I and epidermal growth factor/transforming growth factor alpha. Furthermore, reinitiation of cell cycle progression by insulin/insulin-like growth factor I in hydroxytamoxifen-arrested cells involves dissociation of the corepressors nuclear receptor corepressor (N-CoR) and silencing mediator for retinoid and thyroid hormone receptor (SMRT) from nuclear estrogen receptor alpha and redistribution to the cytoplasm, a process that is inhibited by mitogen-activated protein/extracellular signal-regulated kinase, but not phosphatidylinositol 3'-kinase, inhibitors. These data suggest that agents that up-regulate p27(Kip1) or inhibit growth factor signaling via the extracellular signal-regulated kinases should be tested as therapeutic strategies in tamoxifen-resistant breast cancer. PMID- 12907600 TI - Expulsion of small molecules in vesicles shed by cancer cells: association with gene expression and chemosensitivity profiles. AB - Anticancer drug resistance results from selective pressure of chemotherapy, together with mutations or epigenetic changes that make cells refractory to treatment. In cancer cells, we report that gene expression associated with vesicle shedding correlates with chemosensitivity profiles. Experiments with doxorubicin and other small molecules confirm drug accumulation and expulsion in shed vesicles. Relative differences in the rate of vesicle shedding correspond with doxorubicin resistance across various cell lines. Moreover, accumulation of drug in membrane domains in which vesicles originate accounts for drug expulsion in shed vesicles. These observations implicate vesicle shedding as a drug efflux mechanism potentially involved in drug resistance. PMID- 12907601 TI - Microvascular function regulates intestinal crypt response to radiation. AB - Recent evidence suggests that microvascular endothelial apoptosis represents the primary lesion in radiation damage to the gastrointestinal (GI) tract. Rescue of endothelium by depletion of acid sphingomyelinase or i.v. treatment with basic fibroblast growth factor (FGF) prevented the lethal GI syndrome in C(57)Bl/6 mice. Here we show that basic FGF increased crypt survival after irradiation by 2 3 fold, with a dose modification factor at D(10) of 1.15 (P < 0.01). Basic FGF inhibited initial crypt damage, assessed by crypt shrinkage at 18-24 h, but did not significantly affect the regeneration of surviving crypts at 3.5 days after irradiation. These data suggest that microvascular function regulates expression of radiation-induced crypt stem cell clonogen damage in the evolution of radiation injury to the GI mucosa. PMID- 12907602 TI - Maximum tolerable dose and low-dose metronomic chemotherapy have opposite effects on the mobilization and viability of circulating endothelial progenitor cells. AB - There is growing evidence that vasculogenesis (progenitor cell-derived generation of new blood vessels) is required for the growth of some neoplastic diseases. Here we show that the administration of cyclophosphamide (CTX) at the maximum tolerable dose with 21-day breaks or at more frequent low-dose (metronomic) schedules have opposite effects on the mobilization and viability of circulating endothelial progenitors (CEPs) in immunodeficient mice bearing human lymphoma cells. Animals treated with the maximum tolerable dose CTX experienced a robust CEP mobilization a few days after the end of a cycle of drug administration, and tumors rapidly became drug resistant. Conversely, the administration of metronomic CTX was associated with a consistent decrease in CEP numbers and viability and with more durable inhibition of tumor growth. Our findings suggest that metronomic low-dose chemotherapy regimens are particularly promising for avoiding CEP mobilization and, hence, to potentially reduce vasculogenesis dependent mechanisms of tumor growth. PMID- 12907603 TI - Expression of phosphorylated histone H2AX as a surrogate of cell killing by drugs that create DNA double-strand breaks. AB - Phosphorylation of histone H2AX on serine 139 (gammaH2AX) occurs at sites flanking DNA double-strand breaks and can provide a measure of both number and location of these breaks within the nucleus. Because double-strand breaks are often lethal and are produced by several chemotherapeutic agents, we examined the possibility that expression of gammaH2AX after treatment might be useful as a surrogate indicator of clonogenic cell kill. Chinese hamster V79 cells were exposed for 30 min to drugs known to produce DNA double-stand breaks with different efficiencies: bleomycin, tirapazamine, doxorubicin, etoposide, 4-nitro quinoline-N-oxide, and hydrogen peroxide. Cells were then allowed 1 h to develop foci before fixation or were plated to measure colony formation ability. Anti gammaH2AX antibody staining was measured using flow cytometry. Flow histograms were analyzed for the percentage of cells that showed gammaH2AX levels greater than untreated cells, and this percentage was compared with the clonogenic surviving fraction. H2AX expression measured 1 h after treatment predicted cell killing for all of the drugs examined over two logs of cell kill. Moreover, predictive ability was largely independent of drug type in this cell line, and gammaH2AX levels five times background resulted in 50-90% cell kill. This method seems to provide a useful indicator of clonogenic response to treatment with selected chemotherapeutic drugs. PMID- 12907604 TI - Decreased repair activities of 1,N(6)-ethenoadenine and 3,N(4)-ethenocytosine in lung adenocarcinoma patients. AB - To assess the role of oxidative stress and lipid peroxidation (LPO) in the pathogenesis of lung cancer, we measured the levels of 1,N(6)-ethenoadenine (epsilonA) and 3,N(4)-ethenocytosine (epsilonC) in the DNA by immunoaffinity/(32)P postlabeling (33 cases). We also measured the capacity for epsilonA and epsilonC repair (by the nicking assay) in normal and tumor lung tissues, as well as in blood leukocytes of lung cancer patients (56 cases). Repair activities for epsilonA and epsilonC were also assayed in leukocytes of healthy volunteers, matched with cancer patients for age, sex, and smoking habit (25 individuals). Up to 10-fold variations among individuals were observed both in adducts level and repair activities. No differences in epsilonA and epsilonC levels between tumor and nonaffected lung tissues were recorded. However, leukocytes accumulated a significantly higher number of DNA adducts than the lung tissues. Repair activities for both epsilonA and epsilonC were significantly higher in tumor than in normal lung tissue. No significant differences in epsilonA and epsilonC repair activities were associated with age, sex, or smoking habit. However, a significant difference in repair capacity was observed between two histological types of lung cancer, squamous cell carcinoma (SQ) and adenocarcinoma (AD). In individuals suffering from lung AD, epsilonA- and epsilonC-repair activities in normal lung and blood leukocytes were significantly lower than in SQ patients. Moreover, in nonaffected lung tissue of AD patients, the ratio epsilonA/epsilonC adducts was lower than in SQ patients. Differences have also been found between epsilonA and epsilonC repair activities of cancer patients and healthy volunteers. Repair capacity for epsilonA was significantly lower in blood leukocytes of lung cancer patients than in leukocytes of healthy volunteers (P = 0.012). This difference was even larger between healthy volunteers and patients developing inflammation-related AD (P = 0.00033). Repair activities for epsilonC were the same in leukocytes of healthy controls, all lung cancer patients, and SQ patients. However, individuals with ADs revealed significantly lower epsilonC-repair activity (P = 0.013). These results suggest that oxidative stress-mediated lipid peroxidation might contribute to induction and/or progression of lung cancer. Decreased activity of base excision repair pathway for epsilonA and epsilonC is associated particularly with inflammation related lung AD. PMID- 12907605 TI - Development and characterization of DP-153, a nontumorigenic prostatic cell line that undergoes malignant transformation by expression of dominant-negative transforming growth factor beta receptor type II. AB - We have developed a nontumorigenic epithelial cell line, DP-153, from the dorsal prostate of a Lobund/Wistar rat treated with N-methyl-N-nitrosourea and testosterone propionate. DP-153 cells express cytokeratins 5 and 14, but not cytokeratin 18, consistent with a basal epithelial cell phenotype. Similar to the nontumorigenic NRP-152 prostatic cell line, DP-153 cells do not form tumors in athymic mice and retain many of the properties of normal prostatic cells. They express prostatic acid phosphatase and androgen receptors and require several mitogens (epidermal growth factor, insulin, dexamethasone, and cholera toxin) for sustained growth in culture under serum-containing conditions. DP-153 cells are also growth-stimulated by keratinocyte growth factor and basic fibroblast growth factor and growth-inhibited by all-trans-retinoic acid, 1,25-dihydroxyvitamin D(3), and transforming growth factor (TGF)-beta1. We demonstrate that expression of dominant-negative TGF-beta receptor type II by retroviral transduction of DP 153 cells leads to complete loss of TGF-beta1-induced growth inhibition. When transplanted s.c. in athymic mice, DP-153 cells expressing dominant-negative TGF beta receptor type II form tumors as early as 4 weeks, in contrast to the vector control and parental cell line, which do not form tumors even 8 months after transplantation, supporting the observation that TGF-beta functions as a tumor suppressor in these cells. Our data further support that DP-153 is a suitable cell line for analysis of normal prostatic growth and carcinogenesis. PMID- 12907606 TI - Regulation of caspase expression and apoptosis by adenomatous polyposis coli. AB - The adenomatous polyposis coli (APC) gene, a member of the WNT pathway, has been shown to assign intestinal epithelial cells to a program of proliferation or differentiation through regulation of the beta-catenin/TCF-4 complex. Wild-type APC, in certain cellular contexts, appears to induce differentiation and apoptosis, although mutant forms of APC, known to produce polyps and ultimately cancers, may suppress these events. Here, we show that mutant forms of APC can induce repression of select terminal caspases as a potential means of attenuating responses to apoptotic stimuli. Using gene expression profiling to interrogate the intact intestines of Apc(+/min) mice harboring numerous polyps, we identified a reduction in the mRNA expression of both caspases 3 and 7. We additionally identified a reduction in protein levels of caspase-3, caspase-7, and caspase-9 in human colon cancer specimens known to harbor APC mutations. A reduction in caspase protein levels resulted in resistance to apoptotic-inducing agents and restoration of caspase levels reinstated apoptotic capacities. Consistent with Wnt pathway involvement, dominant negative TCF/LEF induced caspase protein expression. These data provide support for the hypothesis that one of the functions of APC is the regulation of caspase activity and other apoptotic proteins by controlling their expression levels in the cell. PMID- 12907607 TI - Ursolic acid inhibits nuclear factor-kappaB activation induced by carcinogenic agents through suppression of IkappaBalpha kinase and p65 phosphorylation: correlation with down-regulation of cyclooxygenase 2, matrix metalloproteinase 9, and cyclin D1. AB - The process of tumorigenesis requires cellular transformation, hyperproliferation, invasion, angiogenesis, and metastasis. Several genes that mediate these processes are regulated by the transcription factor nuclear factor kappaB (NF-kappaB). The latter is activated by various carcinogens, inflammatory agents, and tumor promoters. Thus, agents that can suppress NF-kappaB activation have the potential to suppress carcinogenesis. Ursolic acid, a pentacyclic triterpene acid, has been shown to suppress the expression of several genes associated with tumorigenesis, but whether ursolic acid mediates its effects through suppression of NF-kappaB is not understood. In the study described in the present report, we found that ursolic acid suppressed NF-kappaB activation induced by various carcinogens including tumor necrosis factor (TNF), phorbol ester, okadaic acid, H(2)O(2), and cigarette smoke. These effects were not cell type specific. Ursolic acid inhibited DNA binding of NF-kappaB consisting of p50 and p65. Ursolic acid inhibited IkappaBalpha degradation, IkappaBalpha phosphorylation, IkappaBalpha kinase activation, p65 phosphorylation, p65 nuclear translocation, and NF-kappaB-dependent reporter gene expression. Ursolic acid also inhibited NF-kappaB-dependent reporter gene expression activated by TNF receptor, TNF receptor-associated death domain, TNF receptor-associated factor, NF-kappaB-inducing kinase, IkappaBalpha kinase, and p65. The inhibition of NF kappaB activation correlated with suppression of NF-kappaB-dependent cyclin D1, cyclooxygenase 2, and matrix metalloproteinase 9 expression. Thus, overall, our results indicate that ursolic acid inhibits IkappaBalpha kinase and p65 phosphorylation, leading to the suppression of NF-kappaB activation induced by various carcinogens. These actions of ursolic acid may mediate its antitumorigenic and chemosensitizing effects. PMID- 12907608 TI - Reduced circulating insulin-like growth factor I levels delay the onset of chemically and genetically induced mammary tumors. AB - Insulin-like growth factors (IGFs) play a crucial role in regulating cell proliferation and differentiation. The aim of this study was to examine the potential relationship between serum IGF-I levels and breast cancer risk. To do this, we studied liver-specific IGF-I gene-deleted (LID) mice, in which circulating IGF-I levels are 25% of that in control mice. Mammary tumors were induced in two ways: (a) by exposing mice to the carcinogen 7,12-dimethybenz (a)anthracene; and (b) by crossing LID mice with C3(1)/SV40 large T-antigen transgenic mice. In both models, LID mice exhibited a delayed latency period of mammary tumor development. In the 7,12-dimethybenz (a)anthracene-induced mammary tumor model, the incidence of palpable mammary tumors was significantly lower in LID mice (26% versus 56% in controls), and the onset of the tumors was delayed (74 +/- 1.2 days in LID mice versus 59.5 +/- 1.1 days in controls). Histological analysis showed extensive squamous metaplasia in late-stage mammary tumors of control mice, whereas late-stage tumors from LID mice exhibited extensive hyperplasia, but little metaplasia. In control mice, the onset of C3(1)/SV40 large T-antigen-induced mammary tumors occurred at 21.6 +/- 1.8 weeks of age, whereas in LID mice the average age of onset was 30.2 +/- 1.7 weeks. In addition, 60% of the mice in the control group developed two or more mammary tumors per mouse, whereas in the LID mice only 30% developed more than one mammary tumor per mouse. Our data demonstrate that circulating IGF-I levels play a significant role as a risk factor in the onset and development of mammary tumors in two well established animal models of breast cancer. PMID- 12907609 TI - Mice with alterations in both p53 and Ink4a/Arf display a striking increase in lung tumor multiplicity and progression: differential chemopreventive effect of budesonide in wild-type and mutant A/J mice. AB - p53 transgenic mice carrying a dominant negative mutation were crossed with Ink4A/Arf heterozygous-deficient mice to investigate whether there is a synergy between these two germ-line mutations in promoting carcinogen-induced lung tumor progression in mice. Mice with a p53 dominant negative mutation and Ink4A/Arf heterozygous deficiency exhibited >20-fold increase in tumor volume compared with approximately 4-fold increase in Ink4A/Arf heterozygous-deficient mice and a 9 fold increase in mice with only the p53 dominant negative mutation. The effect of Ink4A/Arf heterozygous deficiency on lung tumor progression occurred late in the carcinogenesis process (>30 weeks after carcinogen treatment). In addition, most of the lung tumors (approximately 80%) from mice with a p53 mutation and deletion of Ink4A/Arf were lung adenocarcinomas. In contrast, lung adenocarcinomas were seen in <10% of the lung tumors from the wild-type mice and approximately 50% of the lung tumors from Ink4a/Arf heterozygous-deficient or p53 mutant mice. These results indicate a significant synergistic interaction between the presence of a mutant p53 transgene and the Ink4A/Arf deletion during lung tumor progression (P < 0.01). The usefulness of this new mouse model in lung cancer chemoprevention was examined. The chemopreventive efficacy of budesonide was examined in wild type mice, mice with Ink4A/Arf heterozygous deficiency, mice with a mutation in the p53 gene, or mice with both a mutation in the p53 gene and deletion in the Ink4A/Arf locus. Mice treated with budesonide displayed an average of 90% inhibition of lung tumor progression in a standard 18-week chemoprevention assay, regardless of p53 and/or Ink4A/Arf status. However, the efficacy of budesonide against lung tumor progression decreased from 94 to 77% (P = 0.07) in mice with alterations in both p53 and Ink4A/Arf in a 40-week chemoprevention assay. Similarly, when mice bearing established lung adenomas were treated with budesonide, genotype-dependent differential effects of budesonide in wild-type and mutant mice were clearly revealed with a 82, 64, 45, and 33% decrease in tumor volume in wild-type mice, p53(+/+)Ink4a/Arf(+/-) mice, p53(+/ )Ink4a/Arf(+/+) mice, and p53(+/-)Ink4a/Arf(+/-), respectively. Thus, mutant mice with alterations in p53 and/or Ink4A/Arf exhibited a significant resistance to chemoprevention by budesonide. Because p53 and Ink4a/Arf mutations are the most prevalent mutations in human lung cancers, the effectiveness of chemopreventive agents on the mutant A/J mice containing alterations with p53 and Ink4a/Arf is the best preclinical estimate of their efficacy in humans. Thus, the mutant A/J mouse model should prove useful for chemoprevention studies. PMID- 12907611 TI - Identification of tamoxifen-DNA adducts in monkeys treated with tamoxifen. AB - The risk of developing endometrial cancer is increased in breast cancer patients treated with tamoxifen (TAM) and in healthy women undergoing TAM chemoprevention. We have detected previously TAM-DNA adducts in the endometrium of women receiving TAM (Shibutani et al., Carcinogenesis, 21: 1461-1467, 2000). To investigate the genotoxic damage induced by TAM in the uterus and other tissues of primates, we gave adult female cynomolgus monkeys six times the human-equivalent dose of TAM (2 mg/kg body weight/day) for 30 days. DNA samples were prepared from the uterus, ovary, liver, kidney, and brain cortex of three TAM-exposed monkeys and one control monkey and were analyzed as coded specimens. To identify the TAM-DNA adducts, we established a new high-performance liquid chromatography gradient system for (32)P-postlabeling/high-performance liquid chromatography analysis, which can resolve the trans- and cis-diastereoisomers of alpha-(N(2) deoxyguanosinyl)TAM (dG-N(2)-TAM), alpha-(N(2)-deoxyguanosinyl)-N-desmethylTAM, and alpha-(N(2)-deoxyguanosinyl)tamoxifen N-oxide. Trans-forms of dG-N(2)-TAM and dG-N(2)-N-desTAM adducts were detected in the livers of all three TAM-fed monkeys at levels of 2.7 adducts/10(8) nucleotides and 1.7 adducts/10(8) nucleotides, respectively. The levels of dG-N(2)-TAM adducts observed in the uterus of one monkey and in the ovaries of two monkeys were approximately 10-fold lower than those observed in the livers. TAM exposure also induced dG-N(2)-TAM adduct in the brain cortex of all three monkeys with a value of 1.5 adducts/10(8) nucleotides. No TAM-DNA adducts were detected in the kidneys or in any tissues obtained from the unexposed monkey. Our results suggest that women receiving TAM may form genotoxic damage in many organs, including the reproductive organs. PMID- 12907610 TI - Induction of apoptosis by caffeine is mediated by the p53, Bax, and caspase 3 pathways. AB - Caffeine is a key component of many popular drinks, especially tea and coffee. Previous reports have shown that caffeine may contribute to the chemopreventive effect of tea in animals. Here, we report that treatment with low concentrations of caffeine induced apoptosis in JB6 Cl41 cells. JB6 Cl41 cells were starved in 0.1% fetal bovine serum/MEM for 72 h and then treated with 50-450 microM caffeine for 24 h. Cells showed the typical DNA laddering pattern and other characteristics of apoptosis. The IC(50) of caffeine on JB6 Cl41 cells was 2.7 mM. Induction of apoptosis by caffeine appeared to be p53-dependent because cells lacking p53 (p53(-/-)) showed no signs of apoptosis after treatment with caffeine. Immunoprecipitation assays and Western blot analysis showed that caffeine induced phosphorylation of p53 at Ser(15) in JB6 Cl41 cells. The same low concentration of caffeine that was effective for inducing phosphorylation of p53 was also shown to increase p53 activation. Expression of Bax, another p53 target, distinctly increased in a time- and dose-dependent manner. Cleaved caspase 3 was also increased in a time- and dose-dependent manner. These data show that a low concentration of caffeine can induce p53-dependent apoptosis in JB6 cells through the Bax and caspase 3 pathways. PMID- 12907612 TI - Insulin-like growth factor-binding protein-3 gene -202 A/C polymorphism is correlated with advanced disease status in prostate cancer. AB - The circulating level of insulin-like growth factor-binding protein-3 (IGFBP-3) is inversely associated with the risk of prostate cancer (PCa) and its progression and may be modulated by the A/C polymorphism at position -202 in the promoter region of IGFBP-3. This study was conducted to evaluate the role of the A/C polymorphism as a genetic modifier in the etiology of PCa and its disease status. The polymorphism was analyzed by a PCR restriction fragment-length polymorphism technique in 307 PCa patients, 221 benign prostatic hyperplasia (BPH) patients, and 227 male controls. No significant difference in the genotype frequency was found between the PCa or BPH patients and controls (PCa versus control, P = 0.316; BPH versus control, P = 0.964). Regarding the tumor stage, the C allele was more frequently observed in patients having tumors with higher stage (P for trend = 0.002). When the PCa patients with localized disease (stage A + B + C) were considered as reference, those with CC and AC genotype had a significantly increased risk of metastatic disease (stage D) compared with those with AA genotype [age-adjusted odds ratio (aOR) = 3.89, 95% confidence interval (CI) = 1.42-10.64, P = 0.008, and aOR = 1.68, 95% CI = 1.01-2.79, P = 0.044, respectively]. The presence of the C allele appeared to be associated with an increased risk of metastatic PCa with a gene dosage effect (aOR = 1.82, 95% CI = 1.23-2.68, P = 0.002). Similarly, significant findings were also observed when PCa patients were compared between those with organ-confined disease (stage A + B) and those with extra-prostatic extension (stage C + D). Furthermore, the C allele was present more frequently in patients with higher tumor grade. In conclusion, the IGFBP-3 -202 A/C polymorphism was not associated with susceptibility to PCa and BPH in Japanese men, but the presence of the C allele may cumulatively increase the risk for tumor metastasis and for having tumors with a biologically more aggressive phenotype. Because of the significant differences in incidence of clinically evident PCa according to racial backgrounds, the conjecture should be further examined in different racial populations. PMID- 12907613 TI - Phosphatidylinositol 3'-kinase is required for growth of mast cells expressing the kit catalytic domain mutant. AB - The Kit receptor tyrosine kinase is critical for the growth and development of hematopoietic cells, germ cells, and the interstitial cells of Cajal. Gain-of function mutations in codon 816 of the catalytic domain of human Kit [codon 814 of murine Kit (mKit)] are found in patients with mastocytosis, leukemia, and germ cell tumors. There are no drugs that inhibit the activity of Kit catalytic domain mutants to a greater extent than wild-type Kit. The objective of this study was to understand the biochemical mechanisms mediating mast cell transformation by this Kit mutant to identify molecular targets for pharmacological intervention. To this end, we examined signaling pathways activated in the murine mast cell line IC2 infected with either wild-type (IC2-mKit) or mutant mKit (IC2 mKit(D814Y)). In this study, we show that mKit(D814Y) is constitutively phosphorylated on tyrosine 719, and this likely results in constitutive association with activated phosphatidylinositol 3'-kinase (PI3K). In vitro growth of IC2-mKit(D814Y) cells is more sensitive to inhibition of PI3K than SCF-induced growth of IC2-mKit cells. s.c. injection of IC2-mKit(D814Y) in syngeneic mice results in mast cell tumors. To determine whether inhibition of PI3K could reduce mKit(D814Y)-mediated tumorigenicity, mice were treated with 1.5 mg/kg wortmannin three times a week. Five weeks after injection of tumor cells, a 75% reduction in tumor weight was observed when wortmannin treatments were initiated 2 days after inoculation with tumor cells. A 66% reduction occurred when treatment was initiated 2 weeks after inoculation. Treatment with wortmannin increased necrosis in the tumors, and this was associated with apoptosis. Interestingly, there was no effect on tumor vasculature. Thus, PI3K is required for survival and growth of the IC2-mKit(D814Y) mast cell line both in vitro and in vivo. These findings may provide insight into designing strategies for treatment of mastocytosis and other diseases associated with mutations in the Kit catalytic domain. PMID- 12907614 TI - Antitumor effect by interleukin-11 receptor alpha-locus chemokine/CCL27, introduced into tumor cells through a recombinant adenovirus vector. AB - In this study, we examined antitumor activity of a mouse CC chemokine ILC/CCL27 and a mouse CX(3)C chemokine fractalkine/CX(3)CL1 in vivo. We generated recombinant adenovirus vectors with a fiber mutation, encoding mILC (Ad-RGD-mILC) and mFKN (Ad-RGD-mFKN). We confirmed tumor cells infected with Ad-RGD-mILC and Ad RGD-mFKN to express and release these chemokines. Tumor rejection experiments in vivo were carried out by inoculating OV-HM cells infected with Ad-RGD-mILC or Ad RGD-mFKN into immunocompetent mice. mILC significantly suppressed the tumor growth, whereas no such significant effect was observed by mFKN. The antitumor activity induced by mILC was T cell dependent, involving both CD4(+) and CD8(+) T cells. Immunohistochemical analysis revealed accumulation of both CD3(+) lymphocytes and NK cells in the tumor tissue transduced with mILC and mFKN. However, there was a significant difference in the distribution of infiltrating cells. Furthermore, mFKN appeared to have an angiogenic activity, which might have masked its tumor suppressive activity. Collectively, ILC/CCL27 may be a good candidate molecule for cancer gene therapy. PMID- 12907615 TI - Mammary carcinoma suppression by cellular retinoic acid binding protein-II. AB - Retinoic acid (RA) modulates cell proliferation, differentiation, and apoptosis, and is used in chemotherapy and chemoprevention in several human cancers. RA exerts its pleiotropic activities by activating the nuclear receptors, retinoic acid receptor (RAR), which, in turn, regulate transcription of multiple target genes. In cells, RA also associates with cellular RA-binding proteins [cellular RA binding proteins (CRABPs)-I and -II]. Recent studies revealed that CRABP-II functions by "channeling" RA to RAR, thereby enhancing the transcriptional activity of the receptor. In search for a biologically meaningful role for CRABP II, we examined its effect on RA-induced growth inhibition in RA-resistant tumors. Stable expression of CRABP-II in mammary carcinoma SC115 cells enabled activation of RAR, considerably sensitized the cells to RA-induced growth inhibition, and dramatically suppressed their tumorigenicity in immunodeficient mice. Similarly, injection of an adenovirus expressing CRABP-II into mammary carcinomas that spontaneously develop in TgN(MMTVneu)202Mul mice resulted in a significant delay in tumor growth and in prolonged survival rates. Remarkably, in both mouse models, administration of exogenous RA had no additional beneficial effect, indicating that endogenous levels of RA are sufficient for maximal tumor suppression on CRABP-II overexpression. The observations reveal that CRABP-II plays a critical role in sensitizing tumors to the growth-suppressive activities of RA in vivo. PMID- 12907616 TI - E1A, E1B double-restricted adenovirus for oncolytic gene therapy of gallbladder cancer. AB - New treatments, such as gene therapy, are necessary for advanced gallbladder cancer (GBC), but little has been studied. Recent studies have introduced mutant adenoviruses (Ads) with either defective E1B-55kD or mutated E1A, focusing on tumor-specific replication, and the results have been promising. To enhance the safety of this approach, we constructed AxdAdB-3, a double-restricted Ad with a mutant E1A and E1B-55kD deletion. We studied the effects of this Ad in vitro and in vivo on GBC, as well as its safety for normal human cells. We compared the replication and cytopathic effects of AxdAdB-3 in several lines of GBC and primary normal cells with those of wild-type Ad or of AxE1AdB, an E1B-55kD deleted Ad. The efficacy in vivo was examined in nude mice with s.c. implanted or i.p. disseminated GBC. AxdAdB-3 replicated in and caused oncolysis of GBC cell lines (TGBC-44TKB and Mz-ChA2) as efficiently as wild-type Ad or AxE1AdB in vitro. By contrast, AxdAdB-3 replicated much less effectively in primary normal cells (e.g., epithelial cells, endothelial cells, and hepatocytes) than in GBC cells and had only mild cytopathic effects, unlike wild-type Ad. Furthermore, cytotoxicity of AxdAdB-3 in normal cells was milder than that of AxE1AdB. AxdAdB 3 significantly (P < 0.01) suppressed the growth of GBC (TGBC-44TKB) xenografts. AxdAdB-3 was also effective in the treatment of mice with peritoneally disseminated GBC (TGBC-44TKB), demonstrating tumor-selective replication and oncolysis that resulted in significantly (P < 0.05) prolonged survival. The present study shows that the E1 double-restricted Ad effectively and selectively replicates in and causes oncolysis of GBC in vitro and in vivo with reduced negative effects on normal cells, suggesting that this approach could be a promising tool for gene therapy of GBC. PMID- 12907617 TI - All-trans-retinoic acid eliminates immature myeloid cells from tumor-bearing mice and improves the effect of vaccination. AB - Tumor-induced immunosuppression is one of the crucial mechanisms of tumor evasion of immune surveillance. It contributes greatly to the failure of cancer vaccines. Immature myeloid cells (ImCs) play an important role in tumor-induced immunosuppression. These cells accumulate in large numbers in tumor-bearing hosts and directly inhibit T-cell functions via various mechanisms. In this study, we tried to eliminate ImCs in an attempt to improve antitumor response. In vivo administration of all-trans-retinoic acid (ATRA) dramatically reduced the presence of ImCs in all tested tumor models. This effect was not because of a direct antitumor effect of ATRA or decreased production of growth factors by tumor cells. Experiments with adoptive transfer demonstrated that ATRA differentiated ImC in vivo into mature dendritic cells, macrophages, and granulocytes. Decreased presence of ImC in tumor-bearing mice noticeably improved CD4- and CD8-mediated tumor-specific immune response. Combination of ATRA with two different types of cancer vaccines in two different tumor models significantly prolonged the antitumor effect of the treatment. These data suggest that elimination of ImC with ATRA may open an opportunity to improve the effect of cancer vaccines. PMID- 12907618 TI - The biological and biochemical effects of CP-654577, a selective erbB2 kinase inhibitor, on human breast cancer cells. AB - Aberrant expression or activity of epidermal growth factor receptor (EGFr) or the closely related p185(erbB2) can promote cell proliferation and survival and thereby contribute to tumorigenesis. Specific antibodies and low molecular-weight tyrosine kinase inhibitors of both proteins are in clinical trials for cancer treatment. CP-654577 is a potent inhibitor selective for p185(erbB2), relative to EGFr tyrosine kinase, and selectively reduces erbB2 autophosphorylation in intact cells. Treatment of SKBr3 human breast cancer cells with CP-654577 reduces the levels of the activated form of mitogen-activated protein kinase, increases the levels of cyclin-dependent kinase inhibitor p27(kip1) and reduces expression of cyclins D and E. These biochemical changes result in a reduced level of phosphorylated retinoblastoma protein and an inhibition of cell-cycle progression at G(1). Apoptosis is triggered in both SKBr3 and another high erbB2-expressing cell line, BT474, by exposure to 1 micro M CP-654577, but this effect is not observed in MCF7 cells that express low erbB2. Levels of activated Akt, an important positive regulator of cell survival, are reduced within 2 h of exposure to 250 nM CP-654577, and this may contribute to the increased apoptosis. These biochemical effects are distinct from those produced by Tarceva, a selective EGFr inhibitor. The antitumor activity of CP-654577 was investigated in athymic mice bearing s.c. tumors from Fischer rat embryo fibroblasts transfected with erbB2. CP-654577 produced a dose-dependent reduction of p185(erbB2) autophosphorylation and inhibited the growth of these tumors. CP-654577 warrants further evaluation in tumors with high expression of p185(erbB2) and may differ from selective EGFr inhibitors or nonselective dual EGFr/erbB2 inhibitors in efficacy and therapeutic index. PMID- 12907619 TI - Novel mechanisms of apoptosis induced by histone deacetylase inhibitors. AB - Histone deacetylase inhibitors (HDACIs) are a new class of chemotherapeutic drugs able to induce tumor cell apoptosis and/or cell cycle arrest; however, the molecular mechanisms underpinning their anticancer effects are poorly understood. Herein, we assessed the apoptotic pathways activated by three HDACIs, suberoylanilide hydroxamic acid, oxamflatin, and depsipeptide. We determined that all three drugs induced the accumulation of cells with a 4n DNA content and apoptosis mediated by the intrinsic apoptotic pathway. HDACI-induced mitochondrial membrane damage and apoptosis were inhibited by overexpression of Bcl-2, but not by the polycaspase inhibitor N-tert-butoxy-carbonyl-Val-Ala-Asp fluoromethylketone (zVAD-fmk). Moreover, induction of a G(1)-S checkpoint through overexpression of p16(INK4A) or suppression of de novo protein synthesis also inhibited HDACI-induced cell death. Proteolytic cleavage of caspase-2, which is poorly inhibited by zVAD-fmk, was concomitant with HDACI-induced death; however, full processing of caspase-2 to the p19 active form was blocked by Bcl-2. Whereas all three drugs induce the activation of the proapoptotic Bcl-2 protein Bid upstream of mitochondrial membrane disruption, Bid cleavage in response to depsipeptide was significantly attenuated by zVAD-fmk. Suberoylanilide hydroxamic acid and oxamflatin could kill both P-glycoprotein (P-gp)(+) MDR cells and their P-gp(-) counterparts, whereas depsipeptide was shown to be a substrate for P-gp and was less effective in killing P-gp(+) cells. These data provide insight into the functional profile of three HDACIs and are important for the development of more rational approaches to chemotherapy, where information regarding the genetic profile of the tumor is matched with the functional profile of a given chemotherapeutic drug to promote favorable clinical responses. PMID- 12907620 TI - Rapamycin inhibits the interleukin 10 signal transduction pathway and the growth of Epstein Barr virus B-cell lymphomas. AB - EBV-infected B-cell lymphomas are a potentially life-threatening complication in bone marrow and solid organ transplant recipients. Immunosuppressive drugs required to prevent allograft rejection also impair anti-EBV T-cell immunity, thereby increasing the risk of EBV-associated disease. Here we demonstrate that the immunosuppressant rapamycin (RAPA) has a strong antiproliferative effect in vitro on B-cell lines derived from organ transplant recipients with EBV associated posttransplant lymphoproliferative disorder (PTLD). Furthermore, RAPA significantly inhibits or delays the growth of solid tumors established from EBV infected B-cell lines in a xenogeneic mouse model of PTLD. RAPA acts via cell cycle arrest, induction of apoptosis, and, most importantly, inhibition of interleukin 10 secretion, a necessary autocrine growth factor. The reduced interleukin 10 production is accompanied by corresponding decreases in the constitutive activation of the growth-promoting transcription factors signal transducer and activator of transcription 1 and 3. Thus, RAPA can limit B-cell lymphoma growth while simultaneously providing immunosuppression to prevent graft rejection in patients who are otherwise at risk for EBV-associated PTLD. Moreover, these findings may have application to other EBV-associated malignancies. PMID- 12907621 TI - Disease stage variation in CD4+ and CD8+ T-cell reactivity to the receptor tyrosine kinase EphA2 in patients with renal cell carcinoma. AB - We have evaluated CD8+ and CD4+ T-cell responses against a new tumor-associated antigen, the receptor tyrosine kinase EphA2, which is broadly expressed in diverse cancer histologies and is frequently overexpressed in advanced stage/metastatic disease. We report herein that EphA2 is overexpressed in renal cell carcinoma (RCC) cell lines and clinical specimens of RCC, and find that the highest levels of EphA2 are consistently found in the most advanced stages of the disease. We identified and synthesized five putative HLA class I-binding and three class II-binding peptides derived from EphA2 that might serve as targets for immune reactivity. Each peptide induced specific, tumor-reactive CD8+ or CD4+T-cell responses as measured using IFN-gamma enzyme-linked immunospot assays. The EphA2 peptides elicited relatively weak responses from CD8+ T cells derived from normal healthy volunteers or from RCC patients with active disease. In marked contrast, immune reactivity to EphA2-derived epitopes was greatly enhanced in CD8+ T cells that had been isolated from patients who were rendered disease free, after surgery. Furthermore, enzyme-linked immunospot analyses demonstrated prominent EphA2-restricted T-helper 1-type CD4+ T cell activity in patients with early stage disease, whereas T-helper 2-type and T regulatory-type responses predominated in patients with more advanced forms of RCC. These data suggest that the immune system of cancer patients actively monitors EphA2-derived epitopes, and that the magnitude and character of T-cell responses to EphA2 epitopes may convey much-needed predictive information about disease stage and outcome. PMID- 12907622 TI - Synergy between chemotherapy and immunotherapy in the treatment of established murine solid tumors. AB - Cytotoxic chemotherapy is generally considered immunosuppressive, with neutropenia and lymphopenia being common adverse side effects. In this context, we have shown previously that the cytidine analogue, gemcitabine, abolishes humoral responses but, in contrast, augments antigen-specific cellular antitumor immunity. This augmentation occurs in the context of increased antigen cross presentation, T lymphocyte expansion, and infiltration of the tumor. Here, we combine an immunotherapy (CD40 ligation using FGK45; 100 micro g; i.p., q2dx3) with gemcitabine (120 microg/gram; i.p.; q3dx5) to treat established solid tumors. This protocol induced long-term cures in < or =80% of mice, and all of the cured mice were resistant to tumor rechallenge. Synergy between the drug and immunotherapy could not be established in vitro and was only seen in the context of tumor cell death. It was associated with an increase in both CD4 and CD8 T cell infiltration of the tumor, but depletion studies clearly showed that CD4 T cells were not a necessary component of the cure. In contrast, CD8 T cells were absolutely required for the success of this treatment regimen. The priming effect of gemcitabine was not limited to debulking, because mice resected to an equivalent, or lesser residual tumor volume did not eradicate tumor with subsequent immunotherapy. This study provides evidence that chemotherapy has the capacity to augment cellular antitumor immunity, a finding with wider implications for the management of treatment-resistant solid tumors. PMID- 12907623 TI - The circulating dendritic cell compartment in patients with chronic lymphocytic leukemia is severely defective and unable to stimulate an effective T-cell response. AB - Chronic lymphocytic leukemia (CLL), the most frequent leukemia in the Western world, is characterized by a profound dysregulation of the host immune system that has a marked impact on the clinical course of the disease. To date, the competence of the circulating dendritic cell (DC) compartment in CLL patients has not been investigated. To address this issue, we sorted DC precursors from the peripheral blood of CLL patients and found a profoundly altered compartment as compared with normal donors. CLL DCs proved a morphologically and phenotypically immature population, lacking the maturation antigen CD83 and the costimulatory molecule CD80, unable to induce a significant proliferative response in allo mixed lymphocyte reaction, with a reduced ability to release interleukin 12 and to drive a type 1 T-cell response. To investigate whether these defects could be ascribed to inhibiting soluble factors released by the leukemic clone, DCs were generated in vitro from normal monocytes in the presence of allogeneic CLL cells. The addition of CLL cells induced similar markers of abnormal maturation and functional impairment with an inhibition in the expression of costimulatory molecules and a reduction of their allo-stimulatory ability. The blocking of interleukin 6 activity was able to revert the inhibition in a proportion of patients. Taken together, these findings indicate that mechanisms of tumor induced DC inhibition are operational in CLL patients, resulting in both maturative and functional defects in the circulating DC compartment, with a potential functional impact in the regulation of in vivo T-cell immune responses. PMID- 12907624 TI - The apoptosis inhibitor protein survivin induces tumor-specific CD8+ and CD4+ T cells in colorectal cancer patients. AB - The identification of tumor-associated antigens expressed by colorectal carcinoma remains one of the major goals for designing novel immunological treatments for this tumor. By using a reverse-immunology approach, we show here that the inhibitor of apoptosis protein, survivin, is immunogenic in colorectal cancer patients. In particular, we found that survivin elicited CD8(+) T cell-mediated responses in peripheral blood or in tumor-associated lymphocytes from patients at different disease stage. Colorectal carcinoma cells were recognized by survivin specific T lymphocytes, and the survivin-specific, class-I HLA-restricted T lymphocytes were fully activated and released interleukin-2 in response to HLA/survivin-peptide complexes expressed by tumor cells. In addition to CD8 mediated responses, survivin specifically stimulated CD4+ T-cell reactivity in peripheral blood lymphocytes from the same patients, thus suggesting that a complete activation of the immune system may occur in response to this antiapoptotic protein. These findings indicate that survivin could be considered a valuable tumor-associated antigen for immune-based clinical approaches in colorectal cancer. PMID- 12907625 TI - Human CD4+ CD25+ regulatory T cells suppress NKT cell functions. AB - CD4+CD25+ regulatory T cells play an important role in peripheral tolerance. These cells have been reported to be capable of suppressing the response of CD4+CD25- T cells in vitro. The depletion of these cells evokes effective immune responses to tumor cells in vivo. In this study, we demonstrate that CD4+CD25+ T cells also suppress all subsets of Valpha24+NKT cells (Valpha24+CD4-CD8- double negative, Valpha24+CD4+, and Valpha24+CD8+) in both proliferation and cytokine production [IFN-gamma, interleukin-4 (IL-4), IL-13, and IL-10]. This suppression is mediated by cell-to-cell contact but not by a humoral factor or the inhibition of antigen-presenting cells. Moreover, the cytotoxic activity of Valpha24+NKT cells against some tumor cell lines is suppressed by CD4+CD25+ T cells. This finding is important in developing an effective immunotherapy for cancer. PMID- 12907626 TI - Identification of interleukin 8 as an inhibitor of tumor necrosis factor-related apoptosis-inducing ligand-induced apoptosis in the ovarian carcinoma cell line OVCAR3. AB - Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is known to trigger apoptosis in many malignant cells. Whereas cancer cells are responsive to TRAIL-induced cell death when used alone or in combination with other agents, normal cells are known to be relatively less sensitive to the ligand, making it a desirable therapeutic compound to target a variety of cancers. TRAIL induces apoptosis through its interaction with its two proapoptotic death receptors (DRs), DR4 and DR5. In addition, it may also bind the decoy receptors (DcRs), DcR1 and DcR2, which lack an intracellular signaling domain, thus negatively regulating TRAIL-induced apoptosis. Previously, it has been shown that interleukin (IL)-8 is elevated in the ascites of patients with ovarian cancer. Therefore, we examined the role that IL-8 may play in modulating sensitivity to TRAIL-mediated apoptosis. We treated the TRAIL-sensitive cell line OVCAR3 with TRAIL over a period of time with or without pretreatment with IL-8. Here we show the novel findings that IL-8 blocks TRAIL-induced cell death and was able to turn the TRAIL-sensitive cell line into a TRAIL-resistant one. We hypothesized that decreased expression of DRs DR4 and DR5 may contribute to TRAIL resistance. Both reverse transcription-PCR and flow cytometry revealed a decrease in DR4 expression after pretreatment of OVCAR3 cells with IL-8. We have also shown that TRAIL was able to induce caspase-8 cleavage in these cells, whereas pretreatment with IL-8 blocked this caspase cleavage. Through array analysis and confirmation with other techniques, we have determined that IL-8 regulates the expression of a member of the mitogen-activated protein kinase superfamily, p38gamma. These findings provide important insights into the modulation of apoptosis by TRAIL and IL-8 in ovarian cancer. The data suggest a potentially important role of IL-8 in protecting ovarian cancer cells from TRAIL-mediated apoptosis and signify a new potential chemotherapeutic target to augment TRAIL therapy. PMID- 12907627 TI - Transcription factor c-Jun activation represses mdr-1 gene expression. AB - Expression of mdr-1 is complex and highly regulated. Several lines of evidence indirectly suggest that transcription factor c-Jun may negatively regulate human mdr-1 gene expression. We recently found that salvicine, a novel topoisomerase II inhibitor, is cytotoxic for multidrug resistance (MDR) tumor cells and down regulates mdr-1 expression in MDR K562/A02 cells. Salvicine also stimulates a significant increase in the level of c-jun mRNA in HL60 cells. This study investigated the relationship between c-Jun activation and down-regulation of mdr 1 expression by salvicine in K562/A02 cells. Reverse-transcription PCR and Western blotting analyses revealed that salvicine suppressed mdr-1 expression in MDR cells and promoted c-jun expression in both MDR and parental K562 cells. Moreover, levels of c-jun expression were enhanced by salvicine before reduction of mdr-1 expression in K562/A02 cells. Furthermore, c-jun antisense oligodeoxynucleotides prevented salvicine-stimulated enhancement of c-Jun protein and reduction of mdr-1 gene expression, but did not affect the increase in c-jun mRNA levels. Salvicine promoted phosphorylation of c-Jun-N-terminal kinase and c Jun protein in MDR K562/A02 and parental K562 cells. Electrophoretic mobility shift assay analysis showed that salvicine enhanced DNA binding activity of transcription factor activator protein 1. Additionally, c-jun antisense oligodeoxynucleotides also inhibited salvicine-induced apoptosis and cytotoxicity in MDR and parental K562 cells. A possible pathway emerges from these results: salvicine stimulates c-Jun-N-terminal kinase phosphorylation and activation, resulting in c-Jun phosphorylation and activation. Activated c-Jun promotes expression of c-jun itself, represses mdr-1 transcription, and triggers pro apoptotic signals, resulting in low mdr-1 expression and cell death. The present results demonstrate that transcription factor c-Jun plays a principal role in down-regulation of mdr-1 expression and induction of apoptosis in salvicine treated human MDR K562/A02 cells, providing new insights into the complicated mechanisms regulating mdr-1 expression. The findings also suggest that c-Jun might be a potential drug target for circumventing tumor MDR. PMID- 12907628 TI - Opposite effects of modifiers of the ApcMin mutation in intestine and mammary gland. AB - Patterns of tumor susceptibility in different organs are widely divergent in mouse strains: one strain may be highly susceptible to tumors in one organ but resistant in another organ, whereas another strain may exhibit the opposite pattern (P. Demant, Semin. Cancer Biol., 3: 159-166, 1992). Therefore, susceptibility to tumors in different organs is assumed to be controlled by different sets of genes. On the other hand, many oncogenes and tumor suppressor genes are mutated in tumors from different organs, indicating that similar tumorigenic pathways operate in various tissues. To obtain insight into the interactions of susceptibility genes with one of such pathways, we compared tumorigenesis in intestine and mammary gland in recombinant congenic strains (RCSs) carrying the Apc(Min) mutation, affecting the Wnt pathway. The presence of Apc(Min) increased considerably the incidence of intestinal and mammary tumors. The individual RCSs differed in the number and latency of Apc(Min)-induced intestinal and mammary tumors and histological type of the latter. Unexpectedly, the strain distribution of susceptibility to the intestinal and mammary tumors in the Apc(Min)-bearing mice was opposite in the RCSs; the strains most susceptible for intestinal tumors were most resistant to mammary tumors and vice versa. This suggests that a set of genes controls the impact of the Apc(Min) mutation in both organs but with opposite effects. Elucidation of the basis of the observed strain differences in organ-specific Wnt pathway-mediated tumorigenesis will help to understand the interactions between germ-line encoded allelic differences in susceptibility genes and the spectrum of somatic mutations in tumor cells. PMID- 12907629 TI - Genome-wide analysis of gene expression associated with MYCN in human neuroblastoma. AB - Molecular mechanisms through which MYCN expression contributes to the malignant phenotype of neuroblastoma are unknown. We performed a genome-widegene expression analysis of 40 well-characterized neuroblastic tumors and 12 cell lines to identify genes and biological pathways associated with MYCN expression. Gene expression was validated by reverse transcription-PCR and immunohistochemistry using tissue arrays. Two hundred twenty-two of 62,839 oligonucleotide probe sets detected expression of genes that were strongly associated with MYCN expression. Differentially expressed genes included examples of known oncogenes, genes associated with neural differentiation, and genes related to cell proliferation. Expression of a subset of these genes was altered after transfection of a neuroblastoma cell line, SK-N-ER, with a MYCN expressing gene construct when protein synthesis was inhibited and have consensus MYCN binding E-box sequences in their promotor regions, suggesting they represent direct targets. Several novel genes/expressed sequences were identified as overexpressed and most likely coamplified with MYCN in a subset of cases. A classification model to identify neuroblastomas with high levels of MYCN expression was developed based on expression profiles. The identification of coexpressed and coamplified genes associated with MYCN overexpression in neuroblastoma suggests biochemical pathways that contribute to the malignant behavior of these tumors and forms a basis for molecular classification. PMID- 12907630 TI - Wnt pathway activation in mesothelioma: evidence of Dishevelled overexpression and transcriptional activity of beta-catenin. AB - Malignant pleural mesothelioma is a relatively uncommon and yet incurable tumor. The pathogenesis of mesothelioma remains poorly understood. This study evaluated the role of Wnt signaling in mesothelioma. Western blot analysis was conducted to confirm the expression of Dishevelled (Dvl) and cytosolic beta-catenin in matched autologous tissue samples (tumor and normal pleura), malignant pleural effusions, and in established mesothelioma cell lines LRK1A, REN and H513. Thirteen of 15 mesotheliomas examined showed consistent overexpression of Dvl and increased cytosolic beta-catenin levels as compared with controls. To evaluate T-cell factor (Tcf)-dependent transcriptional activity of beta-catenin, luciferase assays were conducted. Fresh mesothelioma cells (effusion derived), as well as LRK1A, REN, and H513 cell lines showed a significant fold increase (1.5-2.4-fold, P < 0.01) in Tcf-dependent transcriptional activity of beta-catenin. To evaluate the biological significance of Dvl function in mesothelioma, a PDZ domain deletion mutant (DeltaPDZ-Dvl) was created and stably transfected into LRK1A, REN, and H513. The effect of DeltaPDZ-Dvl on mesothelioma growth was assayed in vitro (colony formation assay in soft agar) and in vivo (s.c. implantation in athymic mice NCRNU-M). In mesothelioma cells tested, DeltaPDZ-Dvl-mediated inhibition of Dvl decreased cytosolic beta-catenin levels, diminished Tcf mediated transcription, and suppressed tumorigenesis of LRK1A and REN in vitro and in vivo. DeltaPDZ-Dvl also down-regulated expression of c-myc in REN and COX 2 in H513. Our data suggest that in malignant pleural mesothelioma, Wnt signaling is activated through Dvl overexpression and downstream signaling through beta catenin. Inhibition of this signaling leads to significant antitumor effects. These results demonstrate Dvl overexpression in human cancer and, specifically, that Wnt signaling plays a role in mesothelioma pathogenesis. These data offer possible new avenues for therapeutic intervention. PMID- 12907632 TI - High prevalence of BRAF gene mutation in papillary thyroid carcinomas and thyroid tumor cell lines. AB - The RAS-RAF-MEK-ERK-MAP kinase pathway mediates the cellular response to extracellular signals that regulate cell proliferation, differentiation, and apoptosis. Mutation of the RAS proto-oncogene occurs in various thyroid neoplasms such as papillary thyroid carcinomas (PTCs), follicular thyroid adenomas and carcinomas. A second genetic alteration frequently involved in PTC is RET/PTC rearrangements. Recent studies have shown that BRAF, which is a downstream signaling molecule of RET and RAS, is frequently mutated in melanomas. This study tests whether BRAF is also mutated in thyroid tumors and cell lines. We analyzed BRAF gene mutation at codon 599 in thyroid tumors using mutant-allele-specific PCR and in 10 thyroid tumor cell lines by DNA sequencing of the PCR-amplified exon 15. We found that BRAF was mutated in 8 of 10 thyroid tumor cell lines, including 2 of 2 papillary carcinoma cell lines, 4 of 5 anaplastic carcinoma cell lines, 1 of 2 follicular carcinoma cell lines, and 1 follicular adenoma cell line. BRAF mutation at codon 599 was detected in 21 of 56 PTC (38%) but not in 18 follicular adenomas and 6 goiters. BRAF mutation occurred in PTC at a significantly higher frequency in male patients than in female patients. To test whether BRAF mutation may cooperate with RET/PTC rearrangements in the oncogenesis of PTC, we tested whether BRAF-mutated PTCs were also positive for RET/PTC rearrangements. Immunohistochemical staining was conducted to evaluate RET/PTC rearrangements by using two different anti-RET antibodies. Surprisingly, we found that a large number of BRAF-mutated PTCs (8 of 21) also expressed RET, indicating that the RET proto-oncogene is rearranged in these BRAF-mutated PTCs. These observations suggest that mutated BRAF gene may cooperate with RET/PTC to induce the oncogenesis of PTC. PMID- 12907631 TI - Interrogating androgen receptor function in recurrent prostate cancer. AB - The early androgen-dependent (AD) phase of prostate cancer is dependent on the androgen receptor (AR). However, it is unclear whether AR is fully functional in recurrent prostate cancer after androgen withdrawal. To address this issue we interrogated AR signaling in AD and recurrent prostate cancer xenografts using molecular imaging, chromatin immunoprecipitation, and immunohistochemistry. In the imaging experiments, an adenovirus bearing a two-step transcriptional activation cassette, which amplifies AR-dependent firefly luciferase reporter gene activity, was injected into tumors implanted into severe combined immunodeficiency mice. A charge-coupled device optical imaging system detected the initial loss and then resumption of AR transcriptional activity in D luciferin-injected mice as tumors transitioned from AD to recurrent growth. The results of chromatin immunoprecipitation and immunohistochemical localization experiments correlated with the Ad two-step transcriptional activation imaging signal. AR localized to the nucleus and bound to the endogenous prostate-specific antigen enhancer in AD tumors but exited the nucleus and dissociated from the enhancer upon castration. However, AR reentered the nucleus and rebound the prostate-specific antigen enhancer as the cancer transitioned into the recurrent phase. Surprisingly, RNA polymerase II and the general factor TFIIB remained bound to the gene throughout the transition. Our data support the concept that AR is fully functional in recurrent cancer and suggest a model by which a poised but largely inactive transcription complex facilitates reactivation by AR at castrate levels of ligand. PMID- 12907633 TI - FUS/ERG gene fusions in Ewing's tumors. AB - Ewing's tumors are rare pediatric neoplasms that are characterized by specific chromosomal translocations and gene rearrangements. All of the fusion genes reported to date in Ewing's tumors juxtapose the EWS gene at 22q12 to an ETS related gene, the most common of which are FLI1 at 11q24 and ERG at 21q22. We present here four cases of Ewing's tumor, which showed no evidence of a EWS gene rearrangement, but instead contained translocations involving 16p11 and 21q22. A rearrangement involving the same chromosome bands, t(16;21)(p11;q22), is found in rare cases of acute myeloid leukemia and fuses the FUS gene at 16p11 to the ERG gene at 21q22. In two of our Ewing's tumor cases, we were able to show at the sequence level that the translocation between chromosomes 16 and 21 similarly results in a FUS/ERG fusion. In one case, exons 1-5 and most of exon 6 of FUS were fused in-frame to exon 9 of ERG; in the other case, FUS exons 1-7 were fused in-frame to ERG exons 8-9. The functional fusion transcript is expected to be expressed from the der(21)t(16;21) derivative. In the two other t(16;21)-positive Ewing's cases, we performed bacterial artificial chromosome fluorescence in situ hybridization analysis on metaphases and interphase nuclei to demonstrate colocalization of bacterial artificial chromosomes containing FUS and ERG genes, also highly suggestive of fusion gene formation. These represent the first four cases where FUS, rather than EWS, is rearranged with an ETS-family transcription factor in Ewing's tumors. Our data provide additional evidence that the transactivation domains of the TET family of RNA-binding proteins (such as EWS and FUS) are interchangeable, and suggests a novel mechanism of oncogenesis in Ewing's tumors. PMID- 12907634 TI - The impact of microsatellite instability on the molecular phenotype of colorectal tumors. AB - Frequent microsatellite instability MSI (MSI-H) occurring in human tumors is characterized by defective DNA mismatch repair and unique clinical features. However, infrequent MSI (MSI-L) has not been attributable to any other defined molecular pathway, and its existence as a biologically distinct category has been challenged. Moreover, the global molecular phenotypes (GMPs) underlying MSI-H, MSI-L, or microsatellite-stable (MSS) tumors have never been evaluated. To evaluate the impact of MSI status on GMP and to determine the importance of MSI relative to other molecular and clinical features, cDNA microarray-derived data from 41 colon cancers were interpreted using principal components analysis. The clinically relevant principal component with the greatest impact on GMP was component 3, which distinguished MSI-H from non-MSI-H (i.e., MSI-L and microsatellite stable) tumors and was designated the MSI-H separator. Notably, MSI-L cancers were also clearly distinguished from non-MSI-L tumors by another principle component, component 10 (the "MSI-L separator"). This second finding validates the existence of MSI-L tumors as a distinct molecular phenotypic category. Thus, both components 3 and 10 reflected different aspects of MSI and helped to establish principal components analysis as a useful tool to identify and characterize distinct biological features of human malignancy. PMID- 12907636 TI - Widespread bimodal intrachromosomal genomic instability in sporadic breast cancers associated with 13q allelic imbalance. AB - Genomic instability is thought to underlie tumor progression in solid tumors, such as breast cancer. Although evidence that the hereditary breast cancer genes, BRCA1 and BRCA2, are involved in DNA repair suggests that genomic instability plays an important role in hereditary breast tumorigenesis, genomic instability remains poorly characterized in sporadic breast cancers. Using a DNA fingerprinting technique, inter-(simple sequence repeat) PCR (inter-SSR PCR), the degree of genomic instability was quantified in 47 sporadic breast cancers compared with matched adjacent normal breast tissues. Almost all sporadic breast cancers show significant genomic instability by inter-SSR PCR. The distribution of this instability is bimodal; 57% of the tumors show fewer changes, whereas 43% show striking genomic alterations. Further analysis of two inter-SSR PCR tumor normal differences revealed a genomic amplification and probable deletion. Thus, inter-SSR PCR can detect chromosomal breakage-related genomic alterations in most sporadic breast cancers. Genomic instability as detected by inter-SSR PCR is not correlated with aneuploidy, suggesting that this technique preferentially detects intrachromosomal alterations. Chromosomal instability in breast cancer can therefore be subdivided into at least two groups: (a) intrachromosomal and (b) gross chromosomal. Allelic imbalance at markers at the 13q13 and retinoblastoma loci (13q) and not at 17q loci was significantly associated with high levels of intrachromosomal instability, suggesting genes at 13q13 and retinoblastoma loci are either selectively targeted or involved in the genesis of genomic instability in sporadic breast cancers. PMID- 12907635 TI - Inactivation of BHD in sporadic renal tumors. AB - Studies of families with Birt-Hogg-Dube syndrome (BHD) have recently revealed protein-truncating mutations in the BHD gene, leading to tumorigenesis of the skin and of different cell types of kidney. To additionally evaluate the role of BHD in kidney tumorigenesis, we studied 39 sporadic renal tumors of different cell types: 7 renal oncocytomas, 9 chromophobe renal cell carcinomas (RCCs), 11 papillary RCCs, and 12 clear cell RCCs. We screened for BHD mutations and identified a novel somatic mutation in exon 13: c.1939_1966delinsT in a papillary RCC. We performed loss of heterozygosity (LOH) analysis on 28 matched normal/tumor sets, of which 10 of 28 (36%) demonstrated LOH: 2 of 6 (33%) chromophobe RCCs, 5 of 6 (83%) papillary RCCs, 3 of 12 (25%) clear cell RCCs, but 0 of 4 renal oncocytomas. BHD promoter methylation status was examined by a methylation-specific PCR assay of all of the tumors. Methylation was detected in 11 of 39 (28%) sporadic renal tumors: 2 of 7 (29%) renal oncocytomas, 1 of 9 (11%) chromophobe RCCs, 4 of 11 (36%) papillary RCCs, and 4 of 12 (33%) clear cell RCCs. Five tumors with methylation also exhibited LOH. Mutation and methylation were absent in 9 kidney cancer cell lines. Our results showed that somatic BHD mutations are rare in sporadic renal tumors. The alternatives, LOH and BHD promoter methylation, are the two possible inactivating mechanisms involved. In conclusion, unlike other hereditary kidney cancer-related genes (i.e., VHL and MET), which are cell type-specific, BHD is involved in the entire spectrum of histological types of renal tumors, suggesting its major role in kidney cancer tumorigenesis. PMID- 12907637 TI - Susceptibility to aflatoxin B1-related primary hepatocellular carcinoma in mice and humans. AB - The genetic basis of disease susceptibility can be studied by several means, including research on animal models and epidemiological investigations in humans. The two methods are infrequently used simultaneously, but their joint use may overcome the disadvantages of either method alone. We used both approaches in an attempt to understand the genetic basis of aflatoxin B(1) (AFB(1))-related susceptibility to hepatocellular carcinoma (HCC). Ingestion of AFB(1) is a major risk factor for HCC in many areas of the world where HCC is common. Whether humans vary in their ability to detoxify the active intermediate metabolite of AFB(1), AFB(1)-exo-8,9-epoxide, is not certain but may explain why all exposed individuals do not develop HCC. To determine whether human variability in detoxification may exist, in a study of 231 HCC cases and 256 controls, we genotyped eleven loci in two families of AFB(1) detoxification genes; the glutathione S-transferases (GSTs) and the epoxide hydrolases (EPHX). After adjustment for multiple comparisons, only one polymorphism in the epoxide hydrolase family 2 locus remained significantly associated with HCC (odds ratio = 2.06, 95% confidence interval = 1.13-3.12). To determine whether additional susceptibility loci exist, we developed a mouse model system to examine AFB(1) induced HCC. Susceptibility of 7-day-old mice from two common inbred strains (C57BL/6J, DBA/2J) was assessed. DBA/2J animals were 3-fold more sensitive to AFB(1)-induced HCC and significantly more sensitive to AFB(1) acute toxicity than were C57BL/6J animals. Analysis of the xenobiotic metabolizing genes in the two strains revealed single nucleotide polymorphisms in three genes, Gsta4, Gstt1, and Ephx1. Although the GSTT1 and EPHX1 loci did not appear to be related to HCC in the total population of the human study, a polymorphism in GSTA4 was significantly related to risk in the male subset. The mouse model also demonstrated that absent or compromised p53 was not necessary for the development of carcinogenesis. These results indicate that the comparison of results from human studies and the AFB(1)-susceptible mouse model may provide new insights into hepatocarcinogenesis. PMID- 12907638 TI - Identification of 5-fluorouracil-inducible target genes using cDNA microarray profiling. AB - The fluoropyrimidine 5-Fluorouracil (5-FU) is widely used in the treatment of cancer. To identify novel downstream mediators of tumor cell response to 5-FU, we used DNA microarray technology to identify genes that are transcriptionally activated by 5-FU treatment in the MCF-7 breast cancer cell line. Of 2400 genes analyzed, 619 were up-regulated by >3-fold. Highly up-regulated genes (>6-fold) with signal intensities of >3000 were analyzed by Northern blot. Genes that were consistently found to be up-regulated were spermine/spermidine acetyl transferase (SSAT), annexin II, thymosin-beta-10, chaperonin-10, and MAT-8. Treatment of MCF 7 cells with the antifolate tomudex and DNA-damaging agent oxaliplatin also resulted in up-regulation of each of these targets. The 5-FU-induced activation of MAT-8, thymosin-beta-10, and chaperonin-10 was abrogated by inactivation of p53 in MCF-7 cells, whereas induction of SSAT and annexin II was significantly reduced in the absence of p53. Moreover, each of these genes contained more than one potential p53-binding site, suggesting that p53 may play an important regulatory role in 5-FU-induced expression of these genes. In addition, we found that basal expression levels of SSAT, annexin II, thymosin beta-10, and chaperonin-10 were increased (by approximately 2-3-fold), and MAT-8 expression dramatically increased (by approximately 10-fold) in a 5-FU-resistant colorectal cancer cell line (H630-R10) compared with the parental H630 cell line, suggesting these genes may be useful biomarkers of resistance. These results demonstrate the potential of DNA microarrays to identify novel genes involved in mediating the response of tumor cells to chemotherapy. PMID- 12907639 TI - Cell cycle-dependent regulation of the Skp2 promoter by GA-binding protein. AB - Skp2 is the F-box protein component of an SCF-type ubiquitin ligase that interacts specifically with p27(Kip1) and thereby promotes its ubiquitylation and degradation. The abundance of Skp2 mRNA oscillates in a cell cycle-dependent manner, being maximal in S and G(2) phases. The regulation of Skp2 transcription was investigated by cloning the promoter region of the mouse gene and determination of its activity in a luciferase reporter assay. Deletion analysis identified a minimal approximately 0.3-kb promoter region with marked transcriptional activity and a 105-bp essential sequence within this region. Electrophoretic mobility shift assays indicated the presence in nuclear extracts of proteins that bind to this sequence. Site-directed mutagenesis revealed that the core binding motif, CACTTCCG, which is similar to that of GA-binding protein (GABP), is essential for Skp2 transcription. "Supershift" analysis indicated that the protein-probe complexes detected by electrophoretic mobility shift assays contain GABP. Endogenous GABP bound to Skp2 promoter element in a cell cycle dependent manner. Furthermore, overexpression of GABPbeta increased Skp2 promoter activity, and suppression of GABPalpha or GABPbeta by a small interfering RNA resulted in the reduction of Skp2 promoter activity. These data suggest that the cell cycle-dependent binding of GABP to the Skp2 promoter plays an important role in the regulation of Skp2 expression and cell cycle progression from G(1) to S phase. PMID- 12907640 TI - Cdc37 enhances proliferation and is necessary for normal human prostate epithelial cell survival. AB - Cdc37 is a co-chaperone protein that recruits several immature client kinases to Hsp90 for proper folding. Cdc37 up-regulation is a common early event in localized human prostate cancer. Although targeted overexpression in mice leads to prostate epithelial cell hyperplasia, the effect of Cdc37 dysregulation in human prostate cells is unclear. In this study, we examine the role of Cdc37 in the growth regulation of normal prostate epithelial cells using a unique human model system. We demonstrate that Cdc37 overexpression drives proliferation, whereas loss of Cdc37 function arrests growth and leads to apoptosis. With increased Cdc37 expression, molecular analysis of Cdc37 client pathways demonstrates enhanced Raf-1 activity, greater Cdk4 levels, and reduced expression of the cyclin-dependent kinase inhibitor p16/CDKN2. To further investigate these downstream pathways, enhanced Raf-1 or Cdk4 activities were selectively induced in human prostate epithelial cells. Raf-1 activation inhibited proliferation and generated an enlarged, flattened morphology. Induction of Cdk4 activity using cyclin D1 overexpression, however, was sufficient to promote proliferation. These data indicate that Cdc37 induces proliferation and is critical for survival in human prostate epithelial cells. These alterations in cell division and survival may be important in the development and progression of early prostate cancer. PMID- 12907641 TI - Aberrant promoter methylation of the transcription factor genes PAX5 alpha and beta in human cancers. AB - Aberrant methylation of 5'CpG islands is a key epigenetic event in many human cancers. A PCR-based technique of methylated CpG island amplification followed by representational difference analysis was used to identify genes methylated in cancer. Two of the CpG islands identified mapped to the 5' untranslated region of the PAX5 alpha and beta genes. These genes, located on chromosome 9p13, are transcribed from two distinct promoters and form two alternative first exons that are subsequently spliced to the common exons 2-10. The resulting splice variants encode two distinct transcription factors important in cell differentiation and embryonic development. Examination of the methylation status of each gene using methylation-specific PCR revealed that both genes are methylated in approximately 65% of breast and lung tumors. Bisulfite sequencing revealed dense methylation patterns within each 5'CpG island, strongly correlating with transcriptional silencing. Expression in cell lines with dense methylation of either the PAX5 alpha or beta promoter region was restored after treatment with the demethylating agent 5-Aza-2'-deoxycytidine. The PAX5 beta gene encodes for the transcription factor B cell-specific activating protein that, in turn, directly regulates CD19, a gene shown to negatively control cell growth. A strong association was observed between PAX5 beta methylation and loss of expression of the CD19 gene demonstrating that inactivation of the PAX5 beta gene likely contributes to neoplastic development by inhibiting growth regulation through effects on CD19 gene expression. Recent studies have demonstrated the importance of PAX5 gene alterations in human cancer. Our results are the first to identify aberrant promoter methylation as a common mechanism for dysregulation of these genes in solid tumors. PMID- 12907642 TI - Pdef expression in human breast cancer is correlated with invasive potential and altered gene expression. AB - Ets transcription factors control multiple biological processes, including cell proliferation, differentiation, apoptosis, angiogenesis, transformation, and invasion. Pdef is an Ets transcription factor originally identified in prostate tissue. We demonstrate that human Pdef is expressed at high levels primarily in tissues with high epithelial cell content, including prostate, colon, and breast. We also determined that Pdef protein is reduced in human invasive breast cancer and is absent in invasive breast cancer cell lines. We next assessed the functional consequences of these observations. Significantly, expression of Pdef in breast cancer cells leads to inhibition of invasion, migration, and growth. Expression of Pdef also results in the down-regulation of urokinase-type plasminogen activator and activation of the promoter of the tumor suppressor gene, MASPIN: Growth-suppressive effects of Pdef expression are mediated in part by a G(0)-G(1) cell cycle arrest associated with elevated p21 levels. Collectively, these results indicate that Pdef loss may alter the expression of genes controlling progression to invasive breast cancer. PMID- 12907643 TI - Calpain-2 as a target for limiting prostate cancer invasion. AB - Mortality and morbidity of prostate cancer result from extracapsular invasion and metastasis. This tumor progression depends on active cell motility. Previous studies have shown that calpain-regulated rear detachment enabling forward locomotion is required for cell migration initiated by growth factor and adhesion receptors. Therefore, we asked whether calpain would be a target for limiting tumor progression, using as our model the PA DU-145 human prostate carcinoma cell line and a highly invasive subline, wild-type DU-145, derived from it. In vitro, the calpain-specific inhibitor CI-I (ALLN) and the preferential-but-less-specific inhibitor leupeptin decreased transmigration of both cell lines across a Matrigel barrier. These calpain inhibitors limited epidermal growth factor-induced motility but did not alter the growth rate of the tumor cells, as expected. Antisense down-regulation of the growth factor-activated calpain-2 (m-calpain) isoform also reduced transmigration and cell motility. These in vitro findings were then buttressed by in vivo studies, in which i.p. DU-145 tumor xenografts were treated with leupeptin. Tumor invasion into the diaphragm was reduced by leupeptin treatment for both the PA and wild-type DU-145 cells (from 1.7 to 0.78 for the parental line and 2.3 to 1.2 for the invasive derivative, respectively). Tumor cells of both types engineered to express calpain-2 antisense constructs also demonstrated a similar 50% reduced invasiveness in vivo. Finally, we found by gene expression survey of 53 human prostate tumors and 23 normal prostates that calpain was not up-regulated in relationship to invasiveness or metastatic activity, consistent with expectation from the biological role of this effector. Taken together, these results strongly suggest that epigenetic activation of calpain plays an important role in the invasion of human prostate cancer and that it can be targeted to reduce tumor progression. PMID- 12907644 TI - Proteomic analysis of intestinal epithelial cells expressing stabilized beta catenin. AB - Aberrant accumulation of beta-catenin protein because of mutation of either the beta-catenin or adenomatous polyposis coli gene plays an essential role in the development of colorectal carcinoma. We established previously a stable clone of the rat small intestinal epithelial cell line IEC6, which is capable of inducing stabilized beta-catenin protein lacking NH(2)-terminal glycogen synthase kinase 3beta phosphorylation site under a strict control of the tetracycline-regulatory system. This clone, IEC6-TetOFF-beta-catenin DeltaN89, shows in vitro polypoid growth on the removal of doxycycline and seems to be an appropriate model for analyzing the molecular mechanisms of early intestinal carcinogenesis. Of >2000 protein spots displayed by newly developed two-dimensional difference gel electrophoresis, 22 were found to be up- or down-regulated on the induction of stabilized beta-catenin. The majority of these proteins fell into two categories: (a) redox-status regulatory proteins and (b) cytoskeleton-associated proteins. Representatively, a key redox-status regulatory protein, manganese superoxide dismutase, up-regulated in IEC6 cells expressing stabilized beta-catenin protein, was overexpressed in adenoma and adenocarcinoma cells of familial adenomatous polyposis patients in parallel with the accumulation of beta-catenin. These results suggest that aberrant accumulation of beta-catenin might contribute to colorectal carcinogenesis by affecting redox status in the mitochondria of intestinal epithelial cells. PMID- 12907645 TI - Macrophage inhibitory cytokine-1 induces the invasiveness of gastric cancer cells by up-regulating the urokinase-type plasminogen activator system. AB - In our search for genes associated with gastric cancer progression, we identified macrophage inhibitory cytokine-1 (MIC-1), a member of the transforming growth factor beta superfamily, as an overexpressed gene in gastric tumor tissues. Expression analysis of MIC-1 in gastric tumor tissues revealed a specific expression in gastric cancer cells, and this expression level was well correlated with invasive potential in various human gastric cancer cell lines. Stable transfection of MIC-1 into SNU-216, a human gastric cancer cell line, significantly increased its invasiveness. The overexpression of MIC-1 into SNU 216 cells significantly increased the activity of urokinase-type plasminogen activator (uPA), and the expressions of uPA and urokinase-type plasminogen activator receptor (uPAR). Similarly, the stimulation of gastric cancer cell lines with purified recombinant MIC-1 dose-dependently increased cell invasiveness, uPA activity, and uPA and uPAR expression. However, MIC-1 did not significantly suppress the proliferation of gastric cancer cell lines. We also found that the stimulation of human gastric cell lines with recombinant MIC-1 strongly induced activation of mitogen-activated protein kinase kinase-1/2 and extracellular signal-regulated kinase-1/2. Additional analysis revealed that PD98059, a selective inhibitor of mitogen-activated protein kinase kinase-1/2, suppressed not only gastric cancer cell invasiveness and uPA activity, but also the mRNA expressions of uPA and uPAR, as induced by recombinant MIC-1. Our results indicate that MIC-1 may contribute to the malignant progression of gastric cancer cells by inducing tumor cell invasion through the up-regulation of the uPA activation system via extracellular signal-regulated kinase-1/2-dependent pathway. PMID- 12907646 TI - Mutations of APC, K-ras, and p53 are associated with specific chromosomal aberrations in colorectal adenocarcinomas. AB - It is widely accepted that both large-scale chromosomal abnormalities and mutation of specific genes, such as APC, K-ras, and/or p53, occur in the majority of colorectal adenocarcinomas. Whether or not a relationship exists between these different forms of genetic abnormalities was previously unknown. Using comparative genomic hybridization and mutational analysis of APC, K-ras, and p53 to evaluate 50 colorectal adenocarcinomas, we have shown that mutation of p53 is significantly associated with gain of 20q, 13q, and 8q and loss of 18q (P = 0.000, 0.02, 0.044, and 0.001, respectively). Conversely, APC mutation did not associate with any of the above-mentioned aberrations but did associate significantly with gain of 7p (P = 0.01). Gain of chromosomal arm 12p, although a less common aberration, was significantly associated with K-ras mutation (P = 0.011). The associations we have described should refine the search for candidate genes underlying chromosomal aberrations and assist in the definition of distinct pathways in colorectal tumorigenesis. PMID- 12907647 TI - Comparison of immunohistochemistry with reverse transcription-PCR for the detection of micrometastatic prostate cancer in lymph nodes. AB - The objective is to compare the performance of immunohistochemistry (IHC) with that of reverse transcription (RT)-PCR for detecting clinically significant micrometastases in histopathologically normal archival pelvic lymph nodes (PLN) removed at radical prostatectomy from men with locally advanced nonmetastatic prostate cancer. We stained 1864 fixed, paraffin-embedded PLNs from 199 pT(3)N(0)M(0) prostate cancer patients for prostate-specific antigen (PSA) and cytokeratin. We also assessed human glandular kallikrein (hK2) expression in a subset of 164 patients. In addition, all PLN specimens were assayed for hK2 mRNA using a previously described RT-PCR assay. PSA and cytokeratin were expressed in the same 13 of 199 (7%) cases; hK2 was expressed in 3 of 164 (2%) cases. PSA/cytokeratin and hK2 expression were associated with cancer involvement of seminal vesicles, higher Gleason sum, and a positive RT-PCR-hK2 assay result. In standard postoperative multivariable models, IHC-PSA/IHC-Cytokeratin or IHC-hK2 was associated with prostate cancer progression, development of distant metastases, and prostate cancer-specific survival. However, when RT-PCR-hK2 assay result was added to the models, it was the sole predictor of clinical outcomes. Although IHC-PSA/IHC-Cytokeratin and IHC-hK2 were more specific for identifying patients who would suffer biochemical progression and develop metastases and who would ultimately die of prostate cancer, RT-PCR-hK2 was more sensitive and accurate. Although IHC for PSA, cytokeratin, and hK2 appear to be more specific methods for detecting biologically and clinically significant prostate cancer micrometastases in histopathologically normal PLN, RT-PCR-hK2 appears to be a more sensitive method that maintained a reasonable specificity. In pT(3)N(0) patients, a positive RT-PCR-hK2 assay result when performed on PLN was the strongest predictor of clinical outcomes after radical prostatectomy. PMID- 12907648 TI - Mannose receptor (MR) and common lymphatic endothelial and vascular endothelial receptor (CLEVER)-1 direct the binding of cancer cells to the lymph vessel endothelium. AB - Although approximately 50% of cancers give rise to metastases via the lymphatic system, the mechanisms mediating this process have remained unknown. In this study, we have investigated the role of two lymphatic endothelial molecules, the mannose receptor (MR) and common lymphatic endothelial and vascular endothelial receptor (CLEVER)-1 in adhesion of malignant cells to the lymphatic endothelium, and analyzed their expression in two clinical series consisting of squamous cell cancers of the head and neck (n = 17) and breast cancers (n = 72). Affinity of the tested head and neck cancer cell lines to the lymphatic endothelium varied greatly, but adhesion of all cell lines was dependent on both the MR and CLEVER 1. Almost all cancer specimens contained peritumoral vessels that expressed CLEVER-1 and MR, and also the intratumoral lymph vessels often expressed them in both tumor types. However, only intratumoral expression of these molecules seems to be essential for metastatic spread to the regional lymph nodes. Only 8 (22%) of the 36 axillary node-negative breast carcinomas expressed the MR on the intratumoral lymph vessels as compared with 16 (50%) of the 32 node-positive carcinomas (P = 0.017), and all eight head and neck carcinoma patients with regional lymph node metastases at diagnosis had tumors that expressed CLEVER-1 on the intratumoral lymph vessels. These data suggest a role for both the MR and CLEVER-1 in directing the traffic of cancer cells within the lymphatic system. PMID- 12907649 TI - Lsh-deficient murine embryonal fibroblasts show reduced proliferation with signs of abnormal mitosis. AB - Genomic hypomethylation and chromosomal instability are frequent characteristics of human cancer cells. Targeted deletion of Lsh leads to a global defect in genomic methylation, and Lsh-deficient mice die at birth with a reduced body weight. Here, we examine the growth pattern of embryonal fibroblasts derived from Lsh-/- mice. The absence of Lsh leads to a severe proliferative defect of fibroblasts with lower saturation density, early signs of senescence, and a lower frequency of immortalization. The impaired growth rate in vitro may be in part responsible for the small size of Lsh-deficient mice. In addition, Lsh-/- fibroblasts accumulated high centrosome numbers, formed multipolar spindles, displayed micronuclei formation, and elevated nuclear DNA content. A similar increase in centrosome abnormalities was observed when wild-type fibroblasts were treated with a DNA-demethylating agent, suggesting that genomic hypomethylation plays an important role in mitotic defects of Lsh-/- murine embryonal fibroblasts, possibly by altering chromatin structure. Because supernumerary centrosomes are a common feature in cancer cells, this Lsh-dependent pathway has the potential to contribute to genetic instability and chromosomal aberrations during tumor progression. PMID- 12907650 TI - Overexpression of vascular endothelial growth factor by MCF-7 breast cancer cells promotes estrogen-independent tumor growth in vivo. AB - Alteration of the phenotype of breast cancers from estrogen-dependent to estrogen independent growth often leads to the failure of antiestrogenic tumor therapies. We report that overexpression of vascular endothelial growth factor (VEGF) by estrogen-dependent MCF-7 breast cancer cells could abolish estrogen-dependent tumor growth in ovariectomized mice. In the absence of estrogen, MCF-7 VEGF expressing tumors with increased vessel density showed growth kinetics similar to, or even greater than, that of parental MCF-7 tumors with estrogen supplementation. Overexpression of VEGF by MCF-7 cells or treatment on parental MCF-7 cells with recombinant VEGF also stimulated cell proliferation in culture. Our data suggest that VEGF stimulation of MCF-7 tumor angiogenesis and growth is mediated by both autocrine and paracrine mechanisms. PMID- 12907651 TI - Loss of protein kinase Calpha expression may enhance the tumorigenic potential of Gli1 in basal cell carcinoma. AB - Activation of the Sonic hedgehog signaling pathway, primarily through mutational inactivation of the PTCH1 gene, is associated with the development of basal cell carcinoma (BCC). Gli1, a member of the Gli family of transcription factors, is expressed in BCC and in transgenic mice targeted expression of Gli1 in basal keratinocytes leads to BCC development. In addition to BCC, previous studies have shown that Gli1 is expressed in the outer root sheath (ORS) of the hair follicle but is absent in interfollicular epidermis. In this study, we have characterized the expression pattern of two protein kinase C (PKC) isoforms expressed in BCC and hair follicles. We have then used reporter assays to investigate the effects of these isoforms on Gli1 transcriptional activity. We report that in BCC sections, PKCalpha but not PKCdelta was weakly expressed in the epidermis, whereas in the hair follicle, PKCalpha was expressed in the ORS and PKCdelta in the inner root sheath. In contrast, neither PKCalpha nor PKCdelta was expressed in BCC tumor islands, although both isoforms were often expressed in the surrounding stroma. In mammalian 293T cells, coexpression of constitutively active PKCalpha reduced the activity of Gli1 in a dose-dependent manner, whereas constitutively active PKCdelta increased the activity of Gli1, although this required higher expression levels. Regulation of mutant Gli1 protein localized exclusively to the nucleus was similar to that of the wild-type protein, indicating that nuclear-cytoplasmic shuttling is not a determinant of Gli1 control by either PKC isoform. Furthermore, PKC regulation of Gli1 did not involve activation of mitogen-activated protein kinase signaling. Finally, we show that exogenous Gli1 does not alter the expression of PKCalpha in human primary keratinocytes, suggesting that loss of this isoform in BCC is not via Hedgehog signaling. As BCCs have been proposed to originate from the ORS, loss of PKCalpha expression may be relevant to tumor formation; this may, in part, be because of the predicted increase in Gli1 transcriptional activity. PMID- 12907652 TI - Suppression of prostate tumor growth by U19, a novel testosterone-regulated apoptosis inducer. AB - Androgens control prostate homeostasis and regulate androgen response genes. Here, we report the identification and characterization of U19, a novel testosterone-regulated apoptosis inducer with tumor suppressive activity. U19 is an evolutionarily conserved protein expressed in many human tissues, with the most abundant expression in the prostate, bone marrow, kidney, and lymph nodes. Overexpression of U19 in 12 surveyed cell lines induced apoptosis, and new protein synthesis is required for apoptosis induction. Expression of U19 in xenograft prostate tumors markedly induced apoptosis and inhibited tumor growth in vivo. Consistent with its tumor-suppressive role, U19 down-regulation was observed in all of the surveyed prostate cancer cell lines and in 19 of 23 clinical human prostate tumor specimens. Loss of heterozygosity analysis revealed U19 allelic loss in 19 of the 23 specimens. Furthermore, two of the specimens had homozygous U19 deletions, and one specimen had hypermethylated U19 promoter, indicating that U19 can be inactivated genetically or epigenetically. These observations suggest that U19 is growth inhibitory and tumor suppressive and that the disruption of androgen-dependent growth inhibition via U19 down-regulation is commonly associated with prostate cancer progression. PMID- 12907653 TI - Effect of longitudinal oxygen gradients on effectiveness of manipulation of tumor oxygenation. AB - The purpose of this study was to test the hypothesis that longitudinal O(2) gradients in tumor affect response to manipulation of oxygenation. Previously we showed that pO(2) is higher on the fascial than the tumor surface of the R3230Ac rat mammary carcinoma when growing in a dorsal skin-fold window chamber, reflecting a longitudinal oxygen gradient. Magnetic resonance angiography verified prior results: the fascial surface has arterioles and higher vascular density than tumor; and the tumor surface has no arterioles. Phosphorescence lifetime imaging was used to measure each surface hypoxic percentage (HP; percentage of pixels < 10 mm Hg) before and after administration of mannitol or glucose (1 g/kg, i.v.) followed by O(2) breathing. The fascial surface had a smaller HP (median = 2.72%) than tumor (median = 27.94%; P = 0.0002) at baseline. HP on the fascial surface was positively correlated with HP on the tumor surface (P = 0.0067). HP decreased on the fascial surface after either sugar + O(2) (mannitol P = 0.03; glucose P = 0.06; combined P = 0.002), but HP did not change on the tumor surface. Therefore, the tumor surface is refractory to improvement in pO(2) with this method. Additional refinements may be needed to improve pO(2) of analogous regions in larger tumors; mechanism-driven suggestions are provided. PMID- 12907654 TI - Apoptosis induction in prostate cancer cells and xenografts by combined treatment with Apo2 ligand/tumor necrosis factor-related apoptosis-inducing ligand and CPT 11. AB - Because apoptosis is deregulated in most cancers, apoptosis-modulating approaches offer an attractive opportunity for clinical therapy of many tumors, including that of the prostate. LNCaP-derived C4-2 human prostate cancer cells are quite resistant to treatment with Apo2 ligand (Apo2L) or tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), when using a nontagged, Zn-bound recombinant trimeric version that is devoid of any exogeneous sequences and therefore least likely to be immunogenic in human patients and that has been optimized for maximum efficacy and minimum toxicity. When combined with the topoisomerase I inhibitor CPT-11 (irinotecan), Apo2L/TRAIL exhibits enhanced apoptotic activity in C4-2 cells cultured in vitro as well as xenografted as tumors in vivo. Apoptosis both in vitro and in vivo was characterized by two major molecular events. First, apoptosis induction was accompanied by changes in expression levels of the Bcl-2 family genes and their products. However, whereas combination treatment applied to in vitro cell culture was characterized by a significant up regulation and activation of Bax and down-regulation of Bcl-xL, the treatment applied to tumors induced Bak and Bcl-xS, whereas Bcl-omega and Bcl-xL were down regulated. Because there are multiple members of the Bcl-2 family (24 members to date), these data indicate that, under different biological conditions, different proteins may be responsible for activating apoptosis and provide evidence for a differential regulation of the multidomain Bcl-2 protein-encoding genes, bax and bak. Increased Bax expression led to its activation, translocation to the mitochondria, and release of cytochrome c. In addition, this combination treatment induced apoptosis through potent activation of caspase-8 and the proapoptotic protein Bid, resulting in activation of effector caspase-3 and cleavage of its cellular target protein, poly(ADP-ribose) polymerase (PARP), events blocked by the pan-caspase inhibitor N-tert-butoxy-carbonyl-Val-Ala-Asp fluoro methylketone (zVAD-fmk). Activation of multiple caspases and PARP cleavage were also observed in the C4-2 tumors treated with doses resulting in effective tumor control at 42 days after Apo2L/TRAIL plus CPT-11 treatment. Down-regulation of Bax by small interference (RNA) (siRNA) in C4-2 cells significantly prevented PARP cleavage and apoptosis. Strikingly, similar experiments in cells stably expressing a dominant-negative death receptor DR5 led to complete ablation of PARP cleavage and apoptosis, indicating the essential role of both mitochondrial and receptor-mediated apoptotic pathways. Our data indicate that the combined treatment of Apo2L/TRAIL and CPT-11 achieves tumor control in prostate cancer tumors through regulation of Bcl-2 family proteins and potent activation of caspases. PMID- 12907655 TI - Alternative splicing disrupts a nuclear localization signal in spleen tyrosine kinase that is required for invasion suppression in breast cancer. AB - Spleen tyrosine kinase (Syk) is a candidate tumor (metastasis) suppressor that is highly expressed in mammary epithelial cells. Loss of Syk expression through promoter hypermethylation is associated with increased invasiveness in a subset of breast cancer. Here, we show that in addition to full-length Syk [Syk(L)], an alternatively spliced variant, Syk(S), is frequently expressed in breast cancer cells. Syk(S) is identical to Syk(L), except that it lacks 23 amino acid residues (deletion) within the interdomain B (IDB) of Syk. We also show that the aberrant expression of Syk(S) occurs frequently in primary breast tumors but never in matched normal mammary tissues, suggesting a contribution of Syk(S) to mammary tumor progression. Expression of Syk(L) suppressed breast cancer cell invasiveness. In contrast, Syk(S) expression did not affect the cell invasion potential. This differential phenotypic response is accompanied by their different subcellular localization. Immunocytochemical studies and nuclear and cytoplasmic fractionation experiments indicated that Syk(L) could enter the nucleus, whereas Syk(S) was located exclusively in the cytoplasm. Five basic residues in deletion were found to be critical in determining Syk(L) nuclear transport and invasion suppression activity; mutations completely excluded Syk(L) from the nucleus and blocked Syk(L)-inducible invasion suppression. Moreover, IDB acted as an autonomous nuclear localization signal to facilitate nuclear transport of a heterologous protein. Thus, the IDB of Syk(L) contains a nuclear localization signal that is responsible for Syk(L) nuclear translocation. The correlation of the nuclear localization and invasion suppression function of Syk(L) indicated that nuclear Syk possesses biological activities associated with tumor suppression in mammary epithelial cells. PMID- 12907656 TI - The role of transforming growth factor alpha in determining growth factor independence. AB - Growth factor independence is a hallmark of malignancy that is attributed to the development of autocrine growth factor loops in cancer cells. However, growth factor-dependent normal cells also exhibit autocrine activity, thus raising the issue of how endogenously produced activity in cancer cells differs in a manner that leads to growth factor independence. We have examined this issue by comparing growth factor-independent HCT116 human colon carcinoma cells with a growth factor-dependent subcompartment of malignant cells designated HCT116b that was isolated from the same patient tumor. Therefore, the development of the growth factor-independent phenotype represents clonal progression within the tumor in vivo. The growth factor independence of HCT116 cells was shown to be dependent on autocrine transforming growth factor (TGF)-alpha activity, yet the isoparental HCT116b subcompartment showed similar levels of TGF-alpha expression as HCT116 when cells were in exponential growth. When both cell lines were growth arrested by nutrient deprivation, HCT116b cells required nutrient replenishment and growth factors for reinitiation of DNA synthesis, whereas HCT116 cells required only nutrient replenishment. In contrast to growth factor-dependent HCT116b cells, the HCT116 cells showed up-regulation of TGF-alpha expression during growth arrest as a result of enhanced transcription. This increased TGF alpha expression in quiescent HCT116 cells was associated with constitutive epidermal growth factor receptor (EGFR) activation in the growth-arrested state, whereas growth-arrested HCT116b cells did not show EGFR activation. TGF-alpha antisense transfection of HCT116 cells showed that EGFR activation was due to increased TGF-alpha expression. Pretreatment of growth-arrested HCT116 cells with AG1478, a selective inhibitor of EGFR tyrosine kinase activity, blocked the reinitiation of DNA synthesis, demonstrating that growth factor independence was due to the increased TGF-alpha expression and EGFR activation of these cells in growth arrest relative to growth factor-dependent HCT116b cells. Importantly, the level of EGFR activation in growth-arrested HCT116 cells was only slightly higher than that of exponential cells, indicating that it was inappropriate EGFR activation in growth arrest rather than the amplitude of activation that generated growth factor independence. PMID- 12907657 TI - Helicobacter pylori increases proteasome-mediated degradation of p27(kip1) in gastric epithelial cells. AB - Helicobacter pylori infection is associated with increased gastric epithelial cell turnover and is a risk factor for noncardia gastric cancer. H. pylori reduces the expression of p27 protein, a cyclin-dependent kinase inhibitor of the G(1) to S-phase cell cycle transition and gastric tumor suppressor gene. Although cell cycle dysregulation associated with decreased p27 may contribute to gastric carcinogenesis, how H. pylori reduces p27 in gastric epithelial cells remains unknown. In the present study, we investigated the mechanisms of the p27 decrease, using AGS and MKN28 gastric epithelial cells cocultured with H. pylori strains under conditions of defined cell cycle distribution. The expression of p27 protein was reduced by H. pylori in a dose- and time-dependent manner. Northern blot and pulse-chase analyses revealed that this reduction was not regulated at a transcriptional level but by accelerated p27 degradation via a proteasome-dependent pathway. Despite up-regulation of the proteasome-dependent degradation of p27 protein, neither threonine 187-phosphorylated p27 nor skp2 (the ubiquitin ligase for p27) were increased. Furthermore, H. pylori impaired p27 ubiquitination and did not increase global proteasomal function. These results indicate that H. pylori increases the degradation of p27 through a proteasomal pathway distinct from the physiological pathway that degrades p27 during cell cycle progression. Putative virulence genes of H. pylori (cagA, cagE, or vacA) played no role in reducing p27 expression. Increased degradation of p27 by H. pylori through a proteasome-dependent, ubiquitin-independent pathway may contribute to the increased risk of gastric cancer associated with chronic H. pylori infection. PMID- 12907658 TI - Correspondence re: J. B. de Kok et al., DD3, A very sensitive and specific marker to detect prostate tumors. Cancer Res., 62: 2695-2698, 2002. PMID- 12907660 TI - Meeting report: Proteases, extracellular matrix, and cancer: an AACR Special Conference in Cancer Research. PMID- 12907661 TI - Energized by Circulation Research over 30 years. PMID- 12907662 TI - United they go: conjunct regulation of aortic antioxidant enzymes during atherogenesis. PMID- 12907663 TI - Fill a Gab(1) in cardiac hypertrophy signaling: search a missing link between gp130 and ERK5 in hypertrophic remodeling in heart. PMID- 12907664 TI - Priority of experimental evidence. PMID- 12907665 TI - Changes in the Ca2+-activated K+ channels of the coronary artery during left ventricular hypertrophy. AB - It has been suggested that impairment of smooth muscle cell (SMC) function by alterations in the Ca2+-activated K+ (KCa) channels accounts for the reduction in coronary reserve during left ventricular hypertrophy (LVH). However, this hypothesis has not been fully investigated. The main goal of this study was to assess whether the properties of KCa channels in coronary SMCs were altered during LVH. In patch-clamp experiments, the whole-cell currents of the KCa channels were reduced during LVH. The unitary current amplitude and open probability for the KCa channels were significantly reduced in LVH patches compared with control patches. The concentration-response curve of the KCa channel to [Ca2+]i was shifted to the right. Inhibition of the KCa channels by tetraethylammonium (TEA) was more pronounced in LVH cells than in control cells. Western blot analysis indicated no differences in KCa channel expression between the control and LVH coronary SM membranes. In contraction experiments, the effect of high K+ concentration on the resting tension of the LVH coronary artery was greater than on that of the control. The effect of TEA on the resting tension of the LVH coronary artery was reduced compared with the effect on the control. Our findings imply a novel mechanism for reduced coronary reserve during LVH. PMID- 12907666 TI - Respiratory sinus arrhythmia: endogenous activation of nicotinic receptors mediates respiratory modulation of brainstem cardioinhibitory parasympathetic neurons. AB - The heart rate increases during inspiration and decreases during expiration. This respiratory sinus arrhythmia (RSA) occurs by modulation of premotor cardioinhibitory parasympathetic neuron (CPN) activity. However, RSA has not been fully characterized in rats, and despite the critical role of CPNs in the generation of RSA, little is known about the mechanisms that mediate this cardiorespiratory interaction. This study demonstrates that RSA in conscious rats is similar to that in other species. The mechanism of RSA was then examined in vitro. Rhythmic inspiratory-related activity was recorded from the hypoglossal rootlet of 700- to 800-microm medullary sections. CPNs were identified by retrograde fluorescent labeling, and neurotransmission to CPNs was examined using patch-clamp electrophysiological techniques. During inspiratory bursts, the frequency of both spontaneous gamma-aminobutyric acidergic (GABAergic) and spontaneous glycinergic synaptic events in CPNs was significantly increased. Focal application of the nicotinic antagonist dihydro-beta-erythroidine in an alpha4beta2-selective concentration (3 micromol/L) abolished the respiratory evoked increase in GABAergic frequency. In contrast, the increase in glycinergic frequency during inspiration was not altered by nicotinic antagonists. Prenatal nicotine exposure exaggerated the increase in GABAergic frequency during inspiration and enhanced GABAergic synaptic amplitude both between and during inspiratory events. Glycinergic synaptic frequency and amplitude were unchanged by prenatal nicotine exposure. This study establishes a neurochemical link between neurons essential for respiration and CPNs, reveals a functional role for endogenous acetylcholine release and the activation of nicotinic receptors in the generation of RSA, and demonstrates that this cardiorespiratory interaction is exaggerated in rats prenatally exposed to nicotine. PMID- 12907667 TI - Modulation of Na,K-ATPase by the gamma subunit: studies with transfected cells and transmembrane mimetic peptides. AB - The enzymatic activity of the Na,K-ATPase, or sodium pump, is modulated by members of the so-called FXYD family of transmembrane proteins. The best characterized member, FXYD2, also referred to as the gamma subunit, has been shown to decrease the apparent Na+ affinity and increase the apparent ATP affinity of the pump. The effect on ATP affinity had been ascribed to the cytoplasmic C-terminal end of the protein, whereas recent observations suggest that the transmembrane (TM) segment of gamma mediates the Na+ affinity effect. Here we use a novel approach involving synthetic transmembrane mimetic peptides to demonstrate unequivocally that the TM domain of gamma effects the shift in apparent Na+ affinity. Specifically, we show that incubation of these peptides with membranes containing alphabeta pumps modulates Na+ affinity in a manner similar to transfected full-length gamma subunit. Using mutated gamma peptides and transfected proteins, we also show that a specific glycine residue, Gly-41, which is associated with a form of familial renal hypomagnesemia when mutated to Arg, is important for this kinetic effect, whereas Gly-35, located on an alternate face of the transmembrane helix, is not. The peptide approach allows for the analysis of mutants that fail to be expressed in a transfected system. PMID- 12907668 TI - HMGA1 co-activates transcription in B cells through indirect association with DNA. AB - The immunoglobulin heavy chain enhancer, or mu enhancer, is required for B cell development. Only the appropriate combination of transcription factors results in B cell-specific enhancer activation. HMGA1 (formerly (HMG-I(Y)) is a proposed co activator of the ETS transcription factors required for mu enhancer activity. HMGA1 associates with the ETS factor PU.1, resulting in changes in PU.1 structure, and enhanced transcriptional synergy with Ets-1 on the mu enhancer in nonlymphoid cells. New data show HMGA1 directly interacts with Ets-1 in addition to PU.1. In vitro HMGA1/Ets-1 interaction facilitates Ets-1/mu enhancer binding in the absence of an HMGA1.Ets-1.DNA complex. To address whether HMGA1 is present in the transcriptionally active mu nucleoprotein complex, we completed DNA pull down assays to detect protein tethering in the context of protein/DNA interaction. Results show that HMGA1 is not tightly associated with mu enhancer DNA through PU.1 or Ets-1, despite strong associations between these proteins in solution. However, chromatin immunoprecipitation assays show HMGA1 associates with the endogenous enhancer in B cells. Furthermore, antisense HMGA1 substantially decreases mu enhancer activity in B cells. Taken together, these data suggest that HMGA1 functions as a transcriptional mu enhancer co-activator in B cells through indirect association with DNA. PMID- 12907669 TI - In vitro polymerization of heparan sulfate backbone by the EXT proteins. AB - Multiple exosotoses is a dominantly inherited bone disorder caused by defects in EXT1 and EXT2, genes encoding glycosyltransferases involved in heparan sulfate chain elongation. Heparan sulfate polymerization occurs by the alternating addition of glucuronic acid and N-acetylglucosamine units to the nonreducing end of the polysaccharide. EXT1 and EXT2 are suggested to be dual glucuronyl/N acetylglucosaminyltransferases, and a heterooligomeric complex of EXT1 and EXT2 (EXT1/2) is considered to be the biological functional polymerization unit. Here, we have investigated the in vitro polymerization capacities of recombinant soluble EXT1, EXT2, and EXT1/2 complex on exogenous oligosaccharide acceptors derived from Escherichia coli K5 capsular polysaccharide. Incubations of recombinant EXT1 or EXT1/2 complex with 3H-labeled oligosaccharide acceptors and the appropriate nucleotide sugars resulted in conversion of the acceptors to higher molecular weight compounds but with different efficacies for EXT1 and EXT1/2. In contrast, incubations with recombinant EXT2 resulted in the addition of a single glucuronic acid but no further polymerization. These results indicate that EXT1 alone and the EXT1/2 heterocomplex can act as heparan sulfate polymerases in vitro without the addition of additional auxiliary proteins. PMID- 12907670 TI - The structure of 4-hydroxybenzoyl-CoA thioesterase from arthrobacter sp. strain SU. AB - The 4-chlorobenzoyl-CoA dehalogenation pathway in certain Arthrobacter and Pseudomonas bacterial species contains three enzymes: a ligase, a dehalogenase, and a thioesterase. Here we describe the high resolution x-ray crystallographic structure of the 4-hydroxybenzoyl-CoA thioesterase from Arthrobacter sp. strain SU. The tetrameric enzyme is a dimer of dimers with each subunit adopting the so called "hot dog fold" composed of six strands of anti-parallel beta-sheet flanked on one side by a rather long alpha-helix. The dimers come together to form the tetramer with their alpha-helices facing outwards. This quaternary structure is in sharp contrast to that previously observed for the 4-hydroxybenzoyl-CoA thioesterase from Pseudomonas species strain CBS-3, whereby the dimers forming the tetramer pack with their alpha-helices projecting toward the interfacial region. In the Arthrobacter thioesterase, each of the four active sites is formed by three of the subunits of the tetramer. On the basis of both structural and kinetic data, it appears that Glu73 is the active site base in the Arthrobacter thioesterase. Remarkably, this residue is located on the opposite side of the substrate-binding pocket compared with that observed for the Pseudomonas enzyme. Although these two bacterial thioesterases demonstrate equivalent catalytic efficiencies, substrate specificities, and metabolic functions, their quaternary structures, CoA-binding sites, and catalytic platforms are decidedly different. PMID- 12907671 TI - The mechanism of p21-activated kinase 2 autoactivation. AB - The p21-activated kinases (PAKs) play an important role in diverse cellular processes. PAK2 is activated by autophosphorylation upon binding of small G proteins such as Cdc42 and Rac in the GTP-bound state. However, the mechanism of PAK2 autophosphorylation in vitro is unclear. In the present study, the kinetic theory of the substrate reaction during modification of enzyme activity has been applied to a study of the autoactivation of PAK2. On the basis of the kinetic equation of the substrate reaction during the autophosphorylation of PAK2, the activation rate constants for the free enzyme and enzyme-substrate complex have been determined. The results indicate that 1) in the presence of Cdc42, PAK2 autophosphorylation is a bipartite mechanism, with the regulatory domain autophosphorylated at multiple residues, whereas activation coincides with autophosphorylation of the catalytic domain at Thr-402; 2) the autophosphorylation reactions in regulatory domain are either a nonlimiting step or not required for activation of enzyme; 3) the autophosphorylation at site Thr 402 on the catalytic domain occurs by an intermolecular mechanism and is required for phosphorylation of exogenous substrates examined; 4) binding of the exogenous protein/peptide substrates at the active site of PAK2 has little or no effect on the autoactivation of PAK2, suggesting that multiple regions of PAK2 are involved in the enzyme-substrate recognition. The present method also provides a novel approach for studying autophosphorylation reactions. Since the experimental conditions used resemble more closely the in vivo situation where the substrate is constantly being turned over while the enzyme is being modified, this new method would be particularly useful when the regulatory mechanisms of the reversible phosphorylation reaction toward certain enzymes are being assessed. PMID- 12907672 TI - Nitric oxide activates diverse signaling pathways to regulate gene expression. AB - Nitric oxide signaling is crucial for effecting long lasting changes in cells, including gene expression, cell cycle arrest, apoptosis, and differentiation. We have determined the temporal order of gene activation induced by NO in mammalian cells and have examined the signaling pathways that mediate the action of NO. Using microarrays to study the kinetics of gene activation by NO, we have determined that NO induces three distinct waves of gene activity. The first wave is induced within 30 min of exposure to NO and represents the primary gene targets of NO. It is followed by subsequent waves of gene activity that may reflect further cascades of NO-induced gene expression. We verified our results using quantitative real time PCR and further validated our conclusions about the effects of NO by using cytokines to induce endogenous NO production. We next applied pharmacological and genetic approaches to determine the signaling pathways that are used by NO to regulate gene expression. We used inhibitors of particular signaling pathways, as well as cells from animals with a deleted p53 gene, to define groups of genes that require phosphatidylinositol 3-kinase, protein kinase C, NF-kappaB, p53, or combinations thereof for activation by NO. Our results demonstrate that NO utilizes several independent signaling pathways to induce gene expression. PMID- 12907673 TI - Two-stage binding of SecA to the bacterial translocon regulates ribosome translocon interaction. AB - The bacterial translocon interacts with both SecA-bound preproteins and nascent chain-ribosome complexes during Sec and signal recognition particle-dependent protein translocation, respectively. In their inactive state, translocons are saturated with ribosomes and SecA protein, reflecting the inherent affinity of these components for one another. We found that SecA and ribosomes are bound simultaneously and noncompetitively to a common set of inactive translocons. Furthermore, we demonstrate that at a later stage in binding, SecA possesses a ribosome-translocon dissociation activity that is coupled to its ATP-dependent membrane insertion and retraction cycle that drives protein translocation. This novel activity is presumably important in the commitment of the translocon to the Sec-dependent pathway. These results also provide a rationale for the compatibility and regulation of multiple protein translocation pathways that each makes distinct demands on a common translocon core. PMID- 12907674 TI - WW domain HECT E3s target Cbl RING finger E3s for proteasomal degradation. AB - Cbl proteins have RING finger-dependent ubiquitin ligase (E3) activity that is essential for down-regulation of tyrosine kinases. Here we establish that two WW domain HECT E3s, Nedd4 and Itch, bind Cbl proteins and target them for proteasomal degradation. This is dependent on the E3 activity of the HECT E3s but not on that of Cbl. Consistent with these observations, in cells expressing the epidermal growth factor receptor, Nedd4 reverses Cbl-b effects on receptor down regulation, ubiquitylation, and proximal events in signaling. Cbl-b also targets active Src for degradation in cells, and Nedd4 similarly reverses Cbl-mediated Src degradation. These findings establish that RING finger E3s can be substrates, not only for autoubiquitylation but also for ubiquitylation by HECT E3s and suggest an additional level of regulation for Cbl substrates including protein tyrosine kinases. PMID- 12907675 TI - Bone marrow transplantation reveals the in vivo expression of the mitochondrial uncoupling protein 2 in immune and nonimmune cells during inflammation. AB - The mitochondrial uncoupling protein 2 (UCP2) is expressed in spleen, lung, intestine, white adipose tissue, and immune cells. Bone marrow transplantation in mice was used to assess the contribution of immune cells to the expression of UCP2 in basal condition and during inflammation. Immune cells accounted for the total amount of UCP2 expression in the spleen, one-third of its expression in the lung, and did not participate in its expression in the intestine. LPS injection stimulated UCP2 expression in lung, spleen, and intestine in both immune and non immune cells. Successive injections of LPS and dexamethasone or N-acetyl-cysteine prevented the induction of UCP2 in all three tissues, suggesting that oxygen free radical generation plays a role in UCP2 regulation. Finally, both previous studies and our data show that there is down-regulation of UCP2 in immune cells during their activation in the early stages of the LPS response followed by an up regulation in UCP2 during the later stages to protect all cells against oxidative stress. PMID- 12907676 TI - Structural dynamics of myoglobin: ligand migration and binding in valine 68 mutants. AB - We have combined Fourier transform infrared/temperature derivative (FTIR-TDS) spectroscopy at cryogenic temperatures and flash photolysis at ambient temperature to examine the effects of polar and bulky amino acid replacements of the highly conserved distal valine 68 in sperm whale myoglobin. In FTIR-TDS experiments, the CO ligand can serve as an internal voltmeter that monitors the local electrostatic field not only at the active site but also at intermediate ligand docking sites. Mutations of residue 68 alter size, shape, and electric field of the distal pocket, especially in the vicinity of the primary docking site (state B). As a consequence, the infrared bands associated with the ligand at site B are shifted. The effect is most pronounced in mutants with large aromatic side chains. Polar side chains (threonine or serine) have only little effect on the peak frequencies. Ligands that migrate toward more remote sites C and D give rise to IR bands with altered frequencies. TDS experiments separate the photoproducts according to their recombination temperatures. The rates and extent of ligand migration among internal cavities at cryogenic temperatures can be used to interpret geminate and bimolecular O2 and CO recombination at room temperature. The kinetics of geminate recombination can be explained by steric arguments alone, whereas both the polarity and size of the position 68 side chain play major roles in regulating bimolecular ligand binding from the solvent. PMID- 12907677 TI - Lipid transfer inhibitor protein defines the participation of high density lipoprotein subfractions in lipid transfer reactions mediated by cholesterol ester transfer protein (CETP). AB - Cholesterol ester transfer protein (CETP) moves triglyceride (TG) and cholesteryl ester (CE) between lipoproteins. CETP has no apparent preference for high (HDL) or low (LDL) density lipoprotein as lipid donor to very low density lipoprotein (VLDL), and the preference for HDL observed in plasma is due to suppression of LDL transfers by lipid transfer inhibitor protein (LTIP). Given the heterogeneity of HDL, and a demonstrated ability of HDL subfractions to bind LTIP, we examined whether LTIP might also control CETP-facilitated lipid flux among HDL subfractions. CETP-mediated CE transfers from [3H]CE VLDL to various lipoproteins, combined on an equal phospholipid basis, ranged 2-fold and followed the order: HDL3 > LDL > HDL2. LTIP inhibited VLDL to HDL2 transfer at one-half the rate of VLDL to LDL. In contrast, VLDL to HDL3 transfer was stimulated, resulting in a CETP preference for HDL3 that was 3-fold greater than that for LDL or HDL2. Long-term mass transfer experiments confirmed these findings and further established that the previously observed stimulation of CETP activity on HDL by LTIP is due solely to its stimulation of transfer activity on HDL3. TG enrichment of HDL2, which occurs during the HDL cycle, inhibited CETP activity by approximately 2-fold and LTIP activity was blocked almost completely. This suggests that LTIP keeps lipid transfer activity on HDL2 low and constant regardless of its TG enrichment status. Overall, these results show that LTIP tailors CETP-mediated remodeling of HDL3 and HDL2 particles in subclass-specific ways, strongly implicating LTIP as a regulator of HDL metabolism. PMID- 12907678 TI - Origin of higher affinity to RNA of the N-terminal RNA-binding domain than that of the C-terminal one of a mouse neural protein, musashi1, as revealed by comparison of their structures, modes of interaction, surface electrostatic potentials, and backbone dynamics. AB - Musashi1 is an RNA-binding protein abundantly expressed in the developing mouse central nervous system. Its restricted expression in neural precursor cells suggests that it is involved in maintenance of the character of progenitor cells. Musashi1 contains two ribonucleoprotein-type RNA-binding domains (RBDs), RBD1 and RBD2, the affinity to RNA of RBD1 being much higher than that of RBD2. We previously reported the structure and mode of interaction with RNA of RBD2. Here, we have determined the structure and mode of interaction with RNA of RBD1. We have also analyzed the surface electrostatic potential and backbone dynamics of both RBDs. The two RBDs exhibit the same ribo-nucleoprotein-type fold and commonly make contact with RNA on the beta-sheet side. On the other hand, there is a remarkable difference in surface electrostatic potential, the beta-sheet of RBD1 being positively charged, which is favorable for binding negatively charged RNA, but that of RBD2 being almost neutral. There is also a difference in backbone dynamics, the central portion of the beta-sheet of RBD1 being flexible, but that of RBD2 not being flexible. The flexibility of RBD1 may be utilized in the recognition process to facilitate an induced fit. Thus, comparative studies have revealed the origin of the higher affinity of RBD1 than that of RBD2 and indicated that the affinity of an RBD to RNA is not governed by its fold alone but is also determined by its surface electrostatic potential and/or backbone dynamics. The biological role of RBD2 with lower affinity is also discussed. PMID- 12907680 TI - Cryo-atomic force microscopy of unphosphorylated and thiophosphorylated single smooth muscle myosin molecules. AB - The purpose of this study was to determine whether steric blockage of one head by the second head of native two-headed myosin was responsible for the inactivity of nonphosphorylated two-headed myosin compared with the high activity of single headed myosin, as suggested on the basis of electron microscopy of two dimensional crystals of heavy meromyosin (Wendt, T., Taylor, D., Messier, T., Trybus, K. M., and Taylor, K. A. (1999) J. Cell Biol. 147, 1385-1390; and Wendt, T., Taylor, D., Trybus, K. M., and Taylor, K. (2001) Proc. Natl. Acad. Sci. U. S. A. 98, 4361-4366). Our earlier cryo-atomic force microscopy (cryo-AFM) (Zhang, Y., Shao, Z., Somlyo, A. P., and Somlyo, A. V. (1997) Biophys. J. 72, 1308-1318) indicates that thiophosphorylation of the regulatory light chain increases the separation of the two heads of a single myosin molecule, but the thermodynamic probability of steric hindrance by strong binding between the two heads was not determined. We now report this probability determined by cryo-AFM of single whole myosin molecules shown to have normal low ATPase activity (0.007 s-1). We found that the thermodynamic probability of the relative head positions of nonphosphorylated myosin was approximately equal between separated heads as compared with closely apposed heads (energy difference of 0.24 kT (where k is a Boltzman constant and T is the absolute temperature)), and thiophosphorylation increased the number of molecules having separated heads (energy advantage of 1.2 kT (where k is a Boltzman constant and I is the absolute temperature)). Our results do not support the suggestion that strong binding of one head to the other stabilizes the blocked conformation against thermal fluctuations resulting in steric blockage that can account for the low activity of nonphosphorylated two headed myosin. PMID- 12907679 TI - Structure-function analysis of the bestrophin family of anion channels. AB - The bestrophins are a newly described family of anion channels unrelated in primary sequence to any previously characterized channel proteins. The human genome codes for four bestrophins, each of which confers a distinctive plasma membrane conductance on transfected 293 cells. Extracellular treatment with methanethiosulfonate ethyltrimethylammonium (MTSET) of a series of substitution mutants that eliminate one or more cysteines from human bestrophin1 demonstrates that cysteine 69 is the single endogenous cysteine responsible for MTSET inhibition of whole-cell current. Cysteines introduced between positions 78-99 and 223-226 are also accessible to external MTSET, with MTSET modification at positions 79, 80, 83, and 90 producing a 2-6-fold increase in whole-cell current. The latter set of four cysteine-substitution mutants define a region that appears to mediate allosteric control of channel activity. Mapping of transmembrane topography by insertion of N-linked glycosylation sites and tobacco etch virus protease cleavage sites provides evidence for cytosolic N and C termini and an unexpected transmembrane topography with at least three extracellular loops that include positions 60-63, 212-227, and 261-267. These experiments provide the first structural analysis of the bestrophin channel family. PMID- 12907681 TI - Synchronous activation of ERK and phosphatidylinositol 3-kinase pathways is required for collagen and extracellular matrix production in keloids. AB - Keloid fibroproliferation appears to be influenced by epithelial-mesenchymal interactions between keloid keratinocytes (KKs) and keloid fibroblasts (KFs). Keloid and normal fibroblasts exhibit accelerated proliferation and collagen I and III production in co-culture with KKs compared with single cell culture or co culture with normal keratinocytes. ERK and phosphatidylinositol 3-kinase (PI3K) pathway activation has been observed in excessively proliferating KFs in co culture with KKs. We hypothesized that ERK and PI3K pathways might be involved in collagen and extracellular matrix production in KFs. To test our hypothesis, four samples of KFs were co-cultured in defined serum-free medium with KKs for 2-5 days. KF cell lysate was subjected to Western blot analysis. Compared with KF single cell culture, phospho-ERK1/2 and downstream phospho-Elk-1 showed up regulation in the co-culture groups, as did phospho-PI3K and phospho-Akt-1, indicating ERK and PI3K pathway activation. Western blotting of the conditioned medium demonstrated increased collagen I-III, laminin beta2, and fibronectin levels. Addition of the MEK1/2-specific inhibitor U0126 or the PI3K-specific inhibitor LY294002 (but not p38 kinase and JNK inhibitors) completely nullified collagen I-III production and significantly decreased laminin beta2 and fibronectin secretion. In the presence of the MEK1/2 or PI3K inhibitor, fibronectin demonstrated changes in molecular mass reflected by faster in-gel migration. These data strongly suggest that synchronous activation of both the ERK and PI3K pathways is essential for collagen I-III and laminin beta2 production. These pathways additionally appear to affect the side chain attachments of fibronectin. Modulation of these pathways may suggest a direction for keloid therapy. PMID- 12907682 TI - A mutated form of steroidogenic factor 1 (SF-1 G35E) that causes sex reversal in humans fails to synergize with transcription factor GATA-4. AB - Steroidogenic factor 1 (SF-1) is a transcription factor belonging to the nuclear receptor superfamily. SF-1 regulates the expression of many genes involved in reproduction, steroidogenesis, and sexual differentiation. An important SF-1 target for male sexual differentiation is the gene encoding the Mullerian inhibiting substance hormone that induces regression of the Mullerian ducts in the developing male embryo. Not long ago, a mutation (G35E) in the human SF-1 gene was identified as the cause of sex reversal and adrenal failure in a phenotypically female but genotypically XY individual. This suggested that the mutated SF-1 protein might interfere with the expression of SF-1 target gene(s) involved in the male sexual differentiation pathway, such as MIS. Surprisingly, the initial biochemical characterization of the SF-1 G35E mutant revealed that it could bind and activate the MIS promoter as efficiently as wild-type SF-1. MIS expression, however, does not rely solely on SF-1 but rather requires the concerted action of several transcription factors including GATA-4. We have previously reported that GATA-4 and SF-1 transcriptionally cooperate to synergistically activate the MIS promoter. Thus, we hypothesized that the phenotype observed with the SF-1 G35E mutation could be explained, at least in part, by a failure and/or a disruption of GATA-4/SF-1 synergism. We found that the SF-1 G35E mutant failed to synergize with GATA-4 despite a direct physical interaction between the two proteins. Interestingly, the SF-1 G35E mutant also disrupted transcriptional synergism between wild-type SF-1 and GATA-4, indicating that it could act as a dominant negative competitor. Thus, our results strengthen the importance of a GATA-4/SF-1 cooperation for MIS transcription and reveal that disruption of this synergism might be responsible for some cases of abnormal sex differentiation in humans. PMID- 12907683 TI - Plasticity of mitochondrial calcium signaling. AB - Evidence is emerging that a quasisynaptic local communication facilitates the calcium signaling between endoplasmic reticulum and mitochondria. However, it remains elusive whether the machinery of mitochondrial calcium signaling displays plasticity similar to the synaptic transmission. Here we studied the relationship between inositol 1,4,5-trisphosphate (IP3)-linked cytosolic [Ca2+] ([Ca2+]c) oscillations and the associated rise in mitochondrial matrix [Ca2+] ([Ca2+]m) in RBL-2H3 mast cells. We observed that the second [Ca2+]c spike is often associated with a larger rise in the [Ca2+]m than the first. It would appear that this phenomenon was not due to a change in the driving force for Ca2+ uptake and therefore must be due to an enhanced Ca2+ permeability of the mitochondrial Ca2+ uptake sites (uniporter). To investigate the activation and deactivation kinetics of the uniporter during IP3 receptor-mediated Ca2+ mobilization, we established novel methods. Using these approaches, we demonstrated that the IP3-induced increase in the permeability of the uniporter lasted longer than the Ca2+ signal. The sustained increase in Ca2+ permeability was bidirectional. Furthermore, the addition of Ca2+ during the decay of the IP3 effect evoked a large further increase in the uniporter permeability. Calmodulin inhibitors did not interfere with the IP3-induced initial activation of the uniporter but inhibited the sustained phase. These results suggest that the uniporter displays a calmodulin mediated facilitation. This plasticity may allow cooperation among sequential IP3 receptor-mediated [Ca2+] transients in the control of calcium signal propagation to the mitochondria. PMID- 12907684 TI - Actin cytoskeletal architecture regulates nitric oxide-induced apoptosis, dedifferentiation, and cyclooxygenase-2 expression in articular chondrocytes via mitogen-activated protein kinase and protein kinase C pathways. AB - Nitric oxide (NO) in articular chondrocytes regulates differentiation, survival, and inflammatory responses by modulating ERK-1 and -2, p38 kinase, and protein kinase C (PKC) alpha and zeta. In this study, we investigated the effects of the actin cytoskeletal architecture on NO-induced dedifferentiation, apoptosis, cyclooxygenase (COX)-2 expression, and prostaglandin E2 production in articular chondrocytes, with a focus on ERK-1/-2, p38 kinase, and PKC signaling. Disruption of the actin cytoskeleton by cytochalasin D (CD) inhibited NO-induced apoptosis, dedifferentiation, COX-2 expression, and prostaglandin E2 production in chondrocytes cultured on plastic or during cartilage explants culture. CD treatment did not affect ERK-1/-2 activation but blocked the signaling events necessary for NO-induced dedifferentiation, apoptosis, and COX-2 expression such as activation of p38 kinase and inhibition of PKCalpha and -zeta. CD also suppressed activation of downstream signaling of p38 kinase and PKC, such as NF kappaB activation, p53 accumulation, and caspase-3 activation, which are necessary for NO-induced apoptosis. NO production in articular chondrocytes caused down-regulation of phosphatidylinositol (PI) 3-kinase and Akt activities. The down-regulation of PI 3-kinase and Akt was blocked by CD treatment, and the CD effects on apoptosis, p38 kinase, and PKCalpha and -zeta were abolished by the inhibition of PI 3-kinase with LY294002. Our results collectively indicate that the actin cytoskeleton mediates NO-induced regulatory effects in chondrocytes by modulating down-regulation of PI 3-kinase and Akt, activation of p38 kinase, and inhibition of PKCalpha and -zeta PMID- 12907685 TI - In vitro heparan sulfate polymerization: crucial roles of core protein moieties of primer substrates in addition to the EXT1-EXT2 interaction. AB - Heparan, the common unsulfated precursor of heparan sulfate (HS) and heparin, is synthesized on the glycosaminoglycan-protein linkage region tetrasaccharide GlcUA Gal-Gal-Xyl attached to the respective core proteins presumably by HS co polymerases encoded by EXT1 and EXT2, the genetic defects of which result in hereditary multiple exostoses in humans. Although both EXT1 and EXT2 exhibit GlcNAc transferase and GlcUA transferase activities required for the HS synthesis, no HS chain polymerization has been demonstrated in vitro using recombinant enzymes. Here we report in vitro HS polymerization. Recombinant soluble enzymes expressed by co-transfection of EXT1 and EXT2 synthesized heparan polymers with average molecular weights greater than 1.7 x 105 using UDP [3H]GlcNAc and UDP-GlcUA as donors on the recombinant glypican-1 core protein and also on the synthetic linkage region analog GlcUA-Gal-O-C2H4NH-benzyloxycarbonyl. Moreover, in our in vitro polymerization system, a part time proteoglycan, alpha thrombomodulin, that is normally modified with chondroitin sulfate served as a polymerization primer for heparan chain. In contrast, no polymerization was achieved with a mixture of individually expressed EXT1 and EXT2 or with acceptor substrates such as N-acetylheparosan oligosaccharides or the linkage region tetrasaccharide-Ser, which are devoid of a hydrophobic aglycon, suggesting the critical requirement of core protein moieties in addition to the interaction between EXT1 and EXT2 for HS polymerization. PMID- 12907686 TI - Endothelin-1 decreases gap junctional intercellular communication by inducing phosphorylation of connexin 43 in human ovarian carcinoma cells. AB - Endothelin-1 (ET-1) is overexpressed in ovarian carcinoma and acts as an autocrine factor selectively through the ETA receptor (ETAR) to promote tumor cell proliferation, survival, neovascularization, and invasiveness. Loss of gap junctional intercellular communication (GJIC) is critical for tumor progression by allowing the cells to escape growth control. Exposure of HEY and OVCA 433 ovarian carcinoma cell lines to ET-1 led to a 50-75% inhibition in intercellular communication and to a decrease in the connexin 43 (Cx43)-based gap junction plaques. To investigate the phosphorylation state of Cx43, ovarian carcinoma cell lysates were immunoprecipitated and transient tyrosine phosphorylation of Cx43 was detected in ET-1-treated cells. BQ 123, a selective ETAR antagonist, blocked the ET-1-induced Cx43 phosphorylation and cellular uncoupling. Gap junction closure was prevented by tyrphostin 25 and by the selective c-Src inhibitor, PP2. Furthermore, the increased Cx43 tyrosine phosphorylation was correlated with ET-1 induced increase of c-Src activity, and PP2 suppressed the ET-1-induced Cx43 tyrosine phosphorylation, indicating that inhibition of Cx43-based GJIC is mainly mediated by the Src tyrosine kinase pathway. In vivo, the inhibition of human ovarian tumor growth in nude mice induced by the potent ETAR antagonist, ABT-627, was associated with a reduction of Cx43 phosphorylation. These findings indicate that the signaling mechanisms involved in GJIC disruption on ovarian carcinoma cells depend on ETAR activation, which leads to the Cx43 tyrosine phosphorylation mediated by c-Src, suggesting that ETAR blockade may contribute to the control of ovarian carcinoma growth and progression also by preventing the loss of GJIC. PMID- 12907687 TI - Chondroitin sulfate synthase-3. Molecular cloning and characterization. AB - Recently, it has become evident that chondroitin sulfate (CS) glycosyltransferases, which transfer glucuronic acid and/or N-acetylgalactosamine residues from each UDP-sugar to the nonreducing terminus of the CS chain, form a gene family. We report here a novel human gene (GenBank trade mark accession number AB086062) that possesses a sequence homologous with the human chondroitin sulfate synthase-1 (CSS1) gene, formerly known as chondroitin synthase. The full length open reading frame consists of 882 amino acids and encodes a typical type II membrane protein. This enzyme contains a beta 3-glycosyltransferase motif and a beta 4-glycosyltransferase motif similar to that found in CSS1. Both the enzymes were expressed in COS-7 cells as soluble proteins, and their enzymatic natures were characterized. Both glucuronyltransferase and N acetylgalactosaminyltransferase activities were observed when chondroitin, CS polymer, and their corresponding oligosaccharides were used as the acceptor substrates, but no polymerization reaction was observed as in the case of CSS1. The new enzyme was thus designated chondroitin sulfate synthase-3 (CSS3). However, the specific activity of CSS3 was much lower than that of CSS1. The reaction products were shown to have a GlcUA beta 1-3GalNAc linkage and a GalNAc beta 1-4GlcUA linkage in the nonreducing terminus of chondroitin resulting from glucuronyltransferase activity and N-acetylgalactosaminyltransferase activity, respectively. Quantitative real time PCR analysis revealed that the transcript level of CSS3 was much lower than that of CSS1, although it was ubiquitously expressed in various human tissues. These results indicate that CSS3 is a glycosyltransferase having both glucuronyltransferase and N acetylgalactosaminyltransferase activities. It may make a contribution to CS biosynthesis that differs from that of CSS1. PMID- 12907688 TI - Processing and localization of ADAMTS-1 and proteolytic cleavage of versican during cumulus matrix expansion and ovulation. AB - ADAMTS-1 (a disintegrin and metalloprotease with thrombospondin motifs-1) is a member of the ADAMTS family of metalloproteases which, together with ADAMTS-4 and ADAMTS-5, has been shown to degrade members of the lectican family of proteoglycans. ADAMTS-1 mRNA is induced in granulosa cells of periovulatory follicles by the luteinizing hormone surge through a progesterone receptor dependent mechanism. Female progesterone receptor knockout (PRKO) mice are infertile primarily due to ovulatory failure and lack the normal periovulatory induction of ADAMTS-1 mRNA. We therefore investigated the protein localization and function of ADAMTS-1 in ovulating ovaries. Antibodies against two specific peptide regions, the pro-domain and the metalloprotease domain of ADAMTS-1, were generated. Pro-ADAMTS-1 of 110 kDa was identified in mural granulosa cells and appears localized to cytoplasmic secretory vesicles. The mature (85-kDa pro domain truncated) form accumulated in the extracellular matrix of the cumulus oocyte complex (COC) during the process of matrix expansion. Each form of ADAMTS 1 protein increased >10-fold after the ovulatory luteinizing hormone surge in wild-type but not PRKO mice. Versican is also localized selectively to the ovulating COC matrix and was found to be cleaved yielding a 70-kDa N-terminal fragment immunopositive for the neoepitope DPEAAE generated by ADAMTS-1 and ADAMTS-4 protease activity. This extracellular processing of versican was reduced in ADAMTS-1-deficient PRKO mouse ovaries. These observations suggest that one function of ADAMTS-1 in ovulation is to cleave versican in the expanded COC matrix and that the anovulatory phenotype of PRKO mice is at least partially due to loss of this function. PMID- 12907689 TI - The NMR structure of the sensory domain of the membranous two-component fumarate sensor (histidine protein kinase) DcuS of Escherichia coli. AB - The structure of the water-soluble, periplasmic domain of the fumarate sensor DcuS (DcuS-pd) has been determined by NMR spectroscopy in solution. DcuS is a prototype for a sensory histidine kinase with transmembrane signal transfer. DcuS belongs to the CitA family of sensors that are specific for sensing di- and tricarboxylates. The periplasmic domain is folded autonomously and shows helices at the N and the C terminus, suggesting direct linking or connection to helices in the two transmembrane regions. The structure constitutes a novel fold. The nearest structural neighbor is the Per-Arnt-Sim domain of the photoactive yellow protein that binds small molecules covalently. Residues Arg107, His110, and Arg147 are essential for fumarate sensing and are found clustered together. The structure constitutes the first periplasmic domain of a two component sensory system and is distinctly different from the aspartate sensory domain of the Tar chemotaxis sensor. PMID- 12907690 TI - Biosynthesis of 3-O-sulfated heparan sulfate: unique substrate specificity of heparan sulfate 3-O-sulfotransferase isoform 5. AB - Heparan sulfate 3-O-sulfotransferase transfers sulfate to the 3-OH position of a glucosamine to generate 3-O-sulfated heparan sulfate (HS), which is a rare component in HS from natural sources. We previously reported that 3-O- sulfotransferase isoform 5 (3-OST-5) generates both an antithrombin-binding site to exhibit anticoagulant activity and a binding site for herpes simplex virus 1 glycoprotein D to serve as an entry receptor for herpes simplex virus. In this study, we characterize the substrate specificity of 3-OST-5 using the purified enzyme. The enzyme was expressed in insect cells using the baculovirus expression approach and was purified by using heparin-Sepharose and 3',5'-ADP- agarose chromatographies. As expected, the purified enzyme generates both an antithrombin binding site and a glycoprotein D binding site. We isolated IdoUA-AnMan3S and IdoUA-AnMan3S6S from nitrous acid-degraded 3-OST-5-modified HS (pH 1.5), suggesting that 3-OST-5 enzyme sulfates the glucosamine residue that is linked to an iduronic acid residue at the nonreducing end. We also isolated a disaccharide with a structure of DeltaUA2S-GlcNS3S and a tetrasaccharide with a structure of DeltaUA2S-GlcNS-IdoUA2S-GlcNH23S6S from heparin lyases-digested 3-OST-5-modified HS. Our results suggest that 3-OST-5 enzyme sulfates both N-sulfated glucosamine and N-unsubstituted glucosamine residues. Taken together, the results indicate that 3-OST-5 has broader substrate specificity than those of 3-OST-1 and 3-OST-3. The unique substrate specificity of 3-OST-5 serves as an additional tool to study the mechanism for the biosynthesis of biologically active HS. PMID- 12907691 TI - Occurrence of a nontypable Helicobacter pylori strain lacking Lewis blood group O antigens and DD-heptoglycan: evidence for the role of the core alpha1,6-glucan chain in colonization. AB - The cell envelope of Helicobacter pylori contains a lipopolysaccharide (LPS) essential for the physical integrity and functioning of the bacterial cell membrane. The O-chain of this LPS frequently expresses type 2 Lewis x (Lex) and Lewis y (Ley) blood group antigens that mimic human gastric mucosal cell-surface glycoconjugates. This article describes the isolation and structural analysis of the LPS from a clinical isolate of H. pylori strain PJ2 that lacks Le antigens but is still capable of colonization. Subsequent composition, methylation, and CE ESMS analyses of LPS revealed its core oligosaccharide structure to be consistent with the previously proposed structural model for H. pylori LPS. In addition, it carries an unusually long side branch alpha1,6-glucan and was devoid of Le O chain polysaccharide. Its ability to colonize the mouse stomach was essentially identical to that of DD-heptoglycan- and Le antigen- producing H. pylori strains. PMID- 12907692 TI - Vaccination-induced protection of lambs against the parasitic nematode Haemonchus contortus correlates with high IgG antibody responses to the LDNF glycan antigen. AB - Lambs respond to vaccination against bacteria and viruses but have a poor immunological response to nematodes. Here we report that they are protected against the parasitic nematode Haemonchus contortus after vaccination with excretory/secretory (ES) glycoproteins using Alhydrogel as an adjuvant. Lambs immunized with ES in Alhydrogel and challenged with 300 L3 larvae/kg body weight had a reduction in cumulative egg output of 89% and an increased percentage protection of 54% compared with the adjuvant control group. Compared to the adjuvant dimethyl dioctadecyl ammonium bromide, Alhydrogel induced earlier onset and significantly higher ES- specific IgG, IgA, and IgE antibody responses. In all vaccinated groups a substantial proportion of the antibody response was directed against glycan epitopes, irrespective of the adjuvant used. In lambs vaccinated with ES in Alhydrogel but not in any other group a significant increase was found in antibody levels against the GalNAcbeta1,4 (Fucalpha1,3)GlcNAc (fucosylated LacdiNAc, LDNF) antigen, a carbohydrate antigen that is also involved in the host defense against the human parasite Schistosoma mansoni. In lambs the LDNF-specific response increased from the first immunization onward and was significantly higher in protected lambs. In addition, an isotype switch from LDNF-specific IgM to IgG was induced that correlated with protection. These data demonstrate that hyporesponsiveness of lambs to H. contortus can be overcome by vaccination with ES glycoproteins in a strong T helper 2 type response-inducing aluminum adjuvant. This combination generated high and specific antiglycan antibody responses that may contribute to the vaccination-induced protection. PMID- 12907694 TI - Chronic hyperplastic candidosis/candidiasis (candidal leukoplakia). AB - Chronic hyperplastic candidosis/candidiasis (CHC; syn. candidal leukoplakia) is a variant of oral candidosis that typically presents as a white patch on the commissures of the oral mucosa. The major etiologic agent of the disease is the oral fungal pathogen Candida predominantly belonging to Candida albicans, although other systemic co-factors, such as vitamin deficiency and generalized immune suppression, may play a contributory role. Clinically, the lesions are symptomless and regress after appropriate antifungal therapy and correction of underlying nutritional or other deficiencies. If the lesions are untreated, a minor proportion may demonstrate dysplasia and develop into carcinomas. This review outlines the demographic features, etiopathogenesis, immunological features, histopathology, and the role of Candida in the disease process. In the final part of the review, newer molecular biological aspects of the disease are considered together with the management protocols that are currently available, and directions for future research. PMID- 12907693 TI - The role of acquired immunity and periodontal disease progression. AB - Our understanding of the pathogenesis in human periodontal diseases is limited by the lack of specific and sensitive tools or models to study the complex microbial challenges and their interactions with the host's immune system. Recent advances in cellular and molecular biology research have demonstrated the importance of the acquired immune system not only in fighting the virulent periodontal pathogens but also in protecting the host from developing further devastating conditions in periodontal infections. The use of genetic knockout and immunodeficient mouse strains has shown that the acquired immune response-in particular, CD4+ T-cells-plays a pivotal role in controlling the ongoing infection, the immune/inflammatory responses, and the subsequent host's tissue destruction. In particular, studies of the pathogen-specific CD4+ T-cell-mediated immunity have clarified the roles of: (i) the relative diverse immune repertoire involved in periodontal pathogenesis, (ii) the contribution of pathogen associated Th1-Th2 cytokine expressions in periodontal disease progression, and (iii) micro-organism-triggered periodontal CD4+ T-cell-mediated osteoclastogenic factor, 'RANK-L', which is linked to the induction of alveolar bone destruction in situ. The present review will focus on some recent advances in the acquired immune responses involving B-cells, CD8+ T-cells, and CD4+ T-cells in the context of periodontal disease progression. New approaches will further facilitate our understanding of their underlying molecular mechanisms that may lead to the development of new treatment modalities for periodontal diseases and their associated complications. PMID- 12907695 TI - Paracoccidioidomycosis of the mouth: an emerging deep mycosis. AB - Oral fungal infections (mycoses) have come into particular prominence since the advent of infection with Human Immunodeficiency Virus (HIV), and recognition of the Acquired Immune Deficiency Syndrome (AIDS), as well as the phenomenal increase in world travel with increased exposure to infections endemic in the tropics. Paracoccidioidomycosis is a rare mycosis worldwide but common in Brazil and some other areas in Latin America. It can be life-threatening and can manifest with a spectrum of clinical presentations, including frequent oral lesions. This paper reviews the more recent information on Paracoccidioidomycosis, emphasizing those areas most relevant in dental science. PMID- 12907696 TI - Update on burning mouth syndrome: overview and patient management. AB - Burning Mouth Syndrome (BMS) is a chronic pain syndrome that mainly affects middle-aged/old women with hormonal changes or psychological disorders. This condition is probably of multifactorial origin, often idiopathic, and its etiopathogenesis remains largely enigmatic. The present paper discusses several aspects of BMS, updates current knowledge, and provides guidelines for patient management. There is no consensus on the diagnosis and classification of BMS. The etiopathogenesis seems to be complex and in a large number of patients probably involves interactions among local, systemic, and/or psychogenic factors. In the remaining cases, new interesting associations have recently emerged between BMS and either peripheral nerve damage or dopaminergic system disorders, emphasizing the neuropathic background in BMS. Based on these recent data, we have introduced the concepts of "primary" (idiopathic) and "secondary" (resulting from identified precipitating factors) BMS, since this allows for a more systematic approach to patient management. The latter starts with a differential diagnosis based on the exclusion of both other orofacial chronic pain conditions and painful oral diseases exhibiting muco-sal lesions. However, the occurrence of overlapping/overwhelming oral mucosal pathologies, such as infections, may cause difficulties in the diagnosis ("complicated BMS"). BMS treatment is still unsatisfactory, and there is no definitive cure. As a result, a multidisciplinary approach is required to bring the condition under better control. Importantly, BMS patients should be offered regular follow-up during the symptomatic periods and psychological support for alleviating the psychogenic component of the pain. More research is necessary to confirm the association between BMS and systemic disorders, as well as to investigate possible pathogenic mechanisms involving potential nerve damage. If this goal is to be achieved, a uniform definition of BMS and strict criteria for its classification are mandatory. PMID- 12907697 TI - Tooth bleaching--a critical review of the biological aspects. AB - Present tooth-bleaching techniques are based upon hydrogen peroxide as the active agent. It is applied directly, or produced in a chemical reaction from sodium perborate or carbamide peroxide. More than 90% immediate success has been reported for intracoronal bleaching of non-vital teeth, and in the period of 1-8 years' observation time, from 10 to 40% of the initially successfully treated teeth needed re-treatment. Cervical root resorption is a possible consequence of internal bleaching and is more frequently observed in teeth treated with the thermo-catalytic procedure. When the external tooth-bleaching technique is used, the first subjective change in tooth color may be observed after 2-4 nights of tooth bleaching, and more than 90% satisfactory results have been reported. Tooth sensitivity is a common side-effect of external tooth bleaching observed in 15% 78% of the patients, but clinical studies addressing the risk of other adverse effects are lacking. Direct contact with hydrogen peroxide induced genotoxic effects in bacteria and cultured cells, whereas the effect was reduced or abolished in the presence of metabolizing enzymes. Several tumor-promoting studies, including the hamster cheek pouch model, indicated that hydrogen peroxide might act as a promoter. Multiple exposures of hydrogen peroxide have resulted in localized effects on the gastric mucosa, decreased food consumption, reduced weight gain, and blood chemistry changes in mice and rats. Our risk assessment revealed that a sufficient safety level was not reached in certain clinical situations of external tooth bleaching, such as bleaching one tooth arch with 35% carbamide peroxide, using several applications per day of 22% carbamide peroxide, and bleaching both arches simultaneously with 22% carbamide peroxide. The recommendation is to avoid using concentrations higher than 10% carbamide peroxide when one performs external bleaching. We advocate a selective use of external tooth bleaching based on high ethical standards and professional judgment. PMID- 12907698 TI - Implants in the medically compromised patient. AB - Dental clinicians are confronted with an increasing number of medically compromised patients who require implant surgery for their oral rehabilitation. However, there are few guidelines on dental implant therapy in this patient category, so that numerous issues regarding pre- and post-operative management remain unclear to the dental clinician. Therefore, the aim of the present review is to offer a critical evaluation of the literature and to provide the clinician with scientifically based data for implant therapy in the medically compromised patient. This review presents the current knowledge regarding the influence of the most common systemic and local diseases on the outcome of dental implant therapy, e.g., abnormalities in bone metabolism, diabetes mellitus, xerostomia, and ectodermal dysplasias. Specific pathophysiologic aspects of the above mentioned diseases as well as their potential implications for implant success are critically appraised. In line with these implications, guidelines for pre- and post-operative management that may assist in the successful implant-supported rehabilitation of this patient category are proposed. PMID- 12907699 TI - Predator diversity hotspots in the blue ocean. AB - Concentrations of biodiversity, or hotspots, represent conservation priorities in terrestrial ecosystems but remain largely unexplored in marine habitats. In the open ocean, many large predators such as tunas, sharks, billfishes, and sea turtles are of current conservation concern because of their vulnerability to overfishing and ecosystem role. Here we use scientific-observer records from pelagic longline fisheries in the Atlantic and Pacific Oceans to show that oceanic predators concentrate in distinct diversity hotspots. Predator diversity consistently peaks at intermediate latitudes (20-30 degrees N and S), where tropical and temperate species ranges overlap. Individual hotspots are found close to prominent habitat features such as reefs, shelf breaks, or seamounts and often coincide with zooplankton and coral reef hotspots. Closed-area models in the northwest Atlantic predict that protection of hotspots outperforms other area closures in safeguarding threatened pelagic predators from ecological extinction. We conclude that the seemingly monotonous landscape of the open ocean shows rich structure in species diversity and that these features should be used to focus future conservation efforts. PMID- 12907700 TI - Rapid evolution of male-biased gene expression in Drosophila. AB - A number of genes associated with sexual traits and reproduction evolve at the sequence level faster than the majority of genes coding for non-sex-related traits. Whole genome analyses allow this observation to be extended beyond the limited set of genes that have been studied thus far. We use cDNA microarrays to demonstrate that this pattern holds in Drosophila for the phenotype of gene expression as well, but in one sex only. Genes that are male-biased in their expression show more variation in relative expression levels between conspecific populations and two closely related species than do female-biased genes or genes with sexually monomorphic expression patterns. Additionally, elevated ratios of interspecific expression divergence to intraspecific expression variation among male-biased genes suggest that differences in rates of evolution may be due in part to natural selection. This finding has implications for our understanding of the importance of sexual dimorphism for speciation and rates of phenotypic evolution. PMID- 12907701 TI - Molecular cloning and functional expression of the first two specific insect myosuppressin receptors. AB - The Drosophila Genome Project database contains the sequences of two genes, CG8985 and CG13803, which are predicted to code for G protein-coupled receptors. We cloned the cDNAs corresponding to these genes and found that their gene structures had not been correctly annotated. We subsequently expressed the coding regions of the two corrected receptor genes in Chinese hamster ovary cells and found that each of them coded for a receptor that could be activated by low concentrations of Drosophila myosuppressin (EC50,4 x 10(-8) M). The insect myosuppressins are decapeptides that generally inhibit insect visceral muscles. Other tested Drosophila neuropeptides did not activate the two receptors. In addition to the two Drosophila myosuppressin receptors, we identified a sequence in the genomic database from the malaria mosquito Anopheles gambiae that also very likely codes for a myosuppressin receptor. To our knowledge, this paper is the first report on the molecular identification of specific insect myosuppressin receptors. PMID- 12907702 TI - Early resistance to cell death and to onset of the mitochondrial permeability transition during hepatocarcinogenesis with 2-acetylaminofluorene. AB - A hallmark of tumorigenesis is resistance to apoptosis. To explore whether resistance to cell death precedes tumor formation, we have studied the short-term effects of the hepatocarcinogen 2-acetylaminofluorene (AAF) on liver mitochondria, on hepatocytes, and on the response to bacterial endotoxin lipopolysaccharide (LPS) in albino Wistar rats. We show that after as early as two weeks of AAF feeding liver mitochondria developed an increased resistance to opening of the permeability transition pore (PTP), an inner membrane channel that is involved in various forms of cell death. Consistent with a mitochondrial adaptive response in vivo, (i) AAF feeding increased the expression of BCL-2 in mitochondria, and (ii) hepatocytes isolated from AAF-fed rats became resistant to PTP-dependent depolarization, cytochrome c release, and cell death, which were instead observed in hepatocytes from rats fed a control diet. AAF-fed rats were fully protected from the hepatotoxic effects of the injection of 20-30 microg of LPS plus 700 mg of d-galactosamine (d-GalN) x kg-1 of body weight, a treatment that in control rats readily caused a large increase of terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling-positive cells in liver cryosections and release of alanine and aspartate aminotransferase into the bloodstream. Treatment with LPS and d-GalN triggered cleavage of BID, a BCL-2 family member, in the livers of both control- and AAF-fed animals, whereas caspase 3 was cleaved only in control-fed animals, indicating that the mitochondrial proapoptotic pathway had been selectively suppressed during AAF feeding. Phenotypic reversion was observed after stopping the carcinogenic diet. These results underscore a key role of mitochondria in apoptosis and demonstrate that regulation of the mitochondrial PTP is altered early during AAF carcinogenesis, which matches, and possibly causes, the increased resistance of hepatocytes to death stimuli in vivo. Both events precede tumor formation, suggesting that suppression of apoptosis may contribute to the selection of a resistant phenotype, eventually increasing the probability of cell progression to the transformed state. PMID- 12907703 TI - Chronic hepatitis associated with GB virus B persistence in a tamarin after intrahepatic inoculation of synthetic viral RNA. AB - Progress in understanding the pathogenesis of hepatitis C virus (HCV) has been slowed by the absence of tractable small animal models. Whereas GB virus B (GBV B, an unclassified flavivirus) shares a phylogenetic relationship and several biologic attributes with HCV, including hepatotropism, it is not known to cause persistent infection, a hallmark of HCV. Here, we document persistent GBV-B infection in one of two healthy tamarins (Saguinus oedipus) inoculated intrahepatically with infectious synthetic RNA. High-titer viremia (108 to 109 genome equivalents per ml) and transiently elevated serum alanine transaminase activities were present from weeks 4 to 12 postinoculation in both animals. However, whereas GBV-B was eliminated from one animal by 20 weeks, the second animal remained viremic (103 to 107 genome equivalents per ml) for >2 years, with alanine transaminase levels becoming elevated again before spontaneous resolution of the infection. A liver biopsy taken late in the course of infection demonstrated hepatitis with periportal mononuclear infiltrates, hepatocellular microvesicular changes, cytoplasmic lipid droplets, and disordered mitochondrial ultrastructure, findings remarkably similar to chronic hepatitis C. GBV-B infected hepatocytes contained numerous small vesicular membranous structures resembling those associated with expression of HCV nonstructural proteins, and sequencing of GBV-B RNA demonstrated a rate of molecular evolution comparable to that of HCV. We conclude that GBV-B is capable of establishing persistent infections in healthy tamarins, a feature that substantially enhances its value as a model for HCV. Mitochondrial structural changes and altered lipid metabolism leading to steatosis are conserved features of the pathogenesis of chronic hepatitis caused by these genetically distinct flaviviruses. PMID- 12907704 TI - Plants reverse warming effect on ecosystem water balance. AB - Models predict that global warming may increase aridity in water-limited ecosystems by accelerating evapotranspiration. We show that interactions between warming and the dominant biota in a grassland ecosystem produced the reverse effect. In a 2-year field experiment, simulated warming increased spring soil moisture by 5-10% under both ambient and elevated CO2. Warming also accelerated the decline of canopy greenness (normalized difference vegetation index) each spring by 11-17% by inducing earlier plant senescence. Lower transpirational water losses resulting from this earlier senescence provide a mechanism for the unexpected rise in soil moisture. Our findings illustrate the potential for organism-environment interactions to modify the direction as well as the magnitude of global change effects on ecosystem functioning. PMID- 12907705 TI - Induction of sensory long-term facilitation in the carotid body by intermittent hypoxia: implications for recurrent apneas. AB - Reflexes from the carotid body have been implicated in cardiorespiratory disorders associated with chronic intermittent hypoxia (CIH). To investigate whether CIH causes functional and/or structural plasticity in the carotid body, rats were subjected to 10 days of recurrent hypoxia or normoxia. Acute exposures to 10 episodes of hypoxia evoked long-term facilitation (LTF) of carotid body sensory activity in CIH-conditioned but not in control animals. The magnitude of sensory LTF depended on the length of CIH conditioning and was completely reversible and unique to CIH, because conditioning with a comparable duration of sustained hypoxia was ineffective. Histological analysis revealed no differences in carotid body morphology between control and CIH animals. Previous treatment with superoxide anion (O2.-) scavenger prevented sensory LTF. In the CIH conditioned animals, carotid body aconitase enzyme activity decreased compared with controls. These observations suggest that increased generation of reactive oxygen species contribute to sensory LTF. In CIH animals, carotid body complex I activity of the mitochondrial electron transport is inhibited, suggesting mitochondria as one source of O2.- generation. These observations demonstrate that CIH induces a previously uncharacterized form of reactive oxygen species dependent, reversible, functional plasticity in carotid body sensory activity. The sensory LTF may contribute to persistent reflex activation of sympathetic nerve activity and blood pressure in recurrent apnea patients experiencing CIH. PMID- 12907706 TI - Repairing the Sickle Cell mutation. II. Effect of psoralen linker length on specificity of formation and yield of third strand-directed photoproducts with the mutant target sequence. AB - Three identical deoxyoligonucleotide third strands with a 3'-terminal psoralen moiety attached by linkers that differ in length (N = 16, 6 and 4 atoms) and structure were examined for their ability to form triplex-directed psoralen photoproducts with both the mutant T residue of the Sickle Cell beta-globin gene and the comparable wild-type sequence in linear duplex targets. Specificity and yield of UVA (365 nm) and visible (419 nm) light-induced photoadducts were studied. The total photoproduct yield varies with the linker and includes both monoadducts and crosslinks at various available pyrimidine sites. The specificity of photoadduct formation at the desired mutant T residue site was greatly improved by shortening the psoralen linker. In particular, using the N-4 linker, psoralen interaction with the residues of the non-coding duplex strand was essentially eliminated, while modification of the Sickle Cell mutant T residue was maximized. At the same time, the proportion of crosslink formation at the mutant T residue upon UV irradiation was much greater for the N-4 linker. The photoproducts formed with the wild-type target were fully consistent with its single base pair difference. The third strand with the N-4 linker was also shown to bind to a supercoiled plasmid containing the Sickle Cell mutation site, giving photoproduct yields comparable with those observed in the linear mutant target. PMID- 12907707 TI - Repairing the Sickle Cell mutation. III. Effect of irradiation wavelength on the specificity and type of photoproduct formed by a 3'-terminal psoralen on a third strand directed to the mutant base pair. AB - Using a psoralen delivery system mediated by a DNA third strand that binds selectively to linear target duplexes immediately downstream from the Sickle Cell beta-globin gene mutation and the comparable wild-type beta-globin gene sequence, the kinetics of formation and yield of psoralen monoadducts and crosslinks with pyrimidine residues at and near the mutant base pair site and its wild-type counterpart were determined. By exploiting irradiation specificities at 300, 365 and 419 nm, it was possible to evaluate the orientation equilibrium of 3'-linked intercalated psoralen and to develop conditions that lead to preferential formation of each type of photoproduct in both the mutant and wild-type sequences. This makes possible the preparation of each type of photoproduct for use as a substrate for DNA repair. In this way, the base pair change(s) that each generates can be established. PMID- 12907708 TI - Identification and analysis of 'extended -10' promoters in Escherichia coli. AB - We have compiled and aligned the DNA sequences of 554 promoter regions from Escherichia coli and analysed the alignment for sequence similarities. We have focused on the similarities and differences between promoters that either do or do not contain an extended -10 element. The distribution of -10 and -35 hexamer element sequences, the range of spacer lengths between these elements and the frequencies of occurrence of different nucleotides, dinucleotides and trinucleotides were investigated. Extended -10 promoters, which contain a 5'-TG 3' element, tend to have longer spacer lengths than promoters that do not. They also tend to show fewer matches to the consensus -35 hexamer element and contain short runs of T residues in the spacer region. We have shown experimentally that the extended -10 5'-TG-3' motif contributes to promoter activity at seven different promoters. The importance of the motif at different promoters is dependent on the sequence of other promoter elements. PMID- 12907709 TI - Rpb7 subunit of RNA polymerase II interacts with an RNA-binding protein involved in processing of transcripts. AB - Rpb4-Rpb7, a dissociable subcomplex of RNA polymerase II (pol II), is required for transcription initiation. To understand the role of Rpb7 in transcription initiation or other processes in transcription, we carried out a two-hybrid screen for proteins that interact with Rpb7 of the fission yeast Schizosaccharomyces pombe. The screen identified the S.pombe homolog of the Saccharomyces cerevisiae Nrd1, an RNA-binding protein implicated in 3' end formation of small nucleolar and small nuclear RNAs transcribed by pol II. The S.pombe protein, named Seb1 for seven binding, was essential for cell viability, and bound directly to Rpb7 in vitro. Saccharomyces cerevisiae Rpb7 also interacted with Nrd1, indicating that the interaction is conserved in evolution. Glu166 and/or Asp167 of S.pombe Rpb7, residues near the C-terminus of the 172 amino acid protein, were found to be important for its interaction with Seb1. Our results suggest that Rpb7 may function to anchor a processing factor to the pol II apparatus, thereby coupling RNA processing to transcription. The role for Rpb7 is consistent with its location in the pol II complex determined by recent structural studies. PMID- 12907710 TI - Insertion of the T3 DNA polymerase thioredoxin binding domain enhances the processivity and fidelity of Taq DNA polymerase. AB - Insertion of the T3 DNA polymerase thioredoxin binding domain (TBD) into the distantly related thermostable Taq DNA polymerase at an analogous position in the thumb domain, converts the Taq DNA polymerase from a low processive to a highly processive enzyme. Processivity is dependent on the presence of thioredoxin. The enhancement in processivity is 20-50-fold when compared with the wild-type Taq DNA polymerase or to the recombinant polymerase in the absence of thioredoxin. The recombinant Taq DNA pol/TBD is thermostable, PCR competent and able to copy repetitive deoxynucleotide sequences six to seven times more faithfully than Taq DNA polymerase and makes 2-3-fold fewer AT-->GC transition mutations. PMID- 12907711 TI - Probing the substrate specificity of Escherichia coli RNase E using a novel oligonucleotide-based assay. AB - Endoribonuclease RNase E has a central role in both processing and decay of RNA in Escherichia coli, and apparently in many other organisms, where RNase E homologs were identified or their existence has been predicted from genomic data. Although the biochemical properties of this enzyme have been already studied for many years, the substrate specificity of RNase E is still poorly characterized. Here, I have described a novel oligonucleotide-based assay to identify specific sequence determinants that either facilitate or impede the recognition and cleavage of RNA by the catalytic domain of the enzyme. The knowledge of these determinants is crucial for understanding the nature of RNA-protein interactions that control the specificity and efficiency of RNase E cleavage and opens new perspectives for further studies of this multi-domain protein. Moreover, the simplicity and efficiency of the proposed assay suggest that it can be a valuable tool not only for the characterization of RNase E homologs but also for the analysis of other site-specific nucleases. PMID- 12907712 TI - Regulation of TCF ETS-domain transcription factors by helix-loop-helix motifs. AB - DNA binding by the ternary complex factor (TCF) subfamily of ETS-domain transcription factors is tightly regulated by intramolecular and intermolecular interactions. The helix-loop-helix (HLH)-containing Id proteins are trans-acting negative regulators of DNA binding by the TCFs. In the TCF, SAP-2/Net/ERP, intramolecular inhibition of DNA binding is promoted by the cis-acting NID region that also contains an HLH-like motif. The NID also acts as a transcriptional repression domain. Here, we have studied the role of HLH motifs in regulating DNA binding and transcription by the TCF protein SAP-1 and how Cdk-mediated phosphorylation affects the inhibitory activity of the Id proteins towards the TCFs. We demonstrate that the NID region of SAP-1 is an autoinhibitory motif that acts to inhibit DNA binding and also functions as a transcription repression domain. This region can be functionally replaced by fusion of Id proteins to SAP 1, whereby the Id moiety then acts to repress DNA binding in cis. Phosphorylation of the Ids by cyclin-Cdk complexes results in reduction in protein-protein interactions between the Ids and TCFs and relief of their DNA-binding inhibitory activity. In revealing distinct mechanisms through which HLH motifs modulate the activity of TCFs, our results therefore provide further insight into the role of HLH motifs in regulating TCF function and how the inhibitory properties of the trans-acting Id HLH proteins are themselves regulated by phosphorylation. PMID- 12907713 TI - A yeast knockout strain to discriminate between active and inactive tRNA molecules. AB - Here we report the construction of a yeast genetic screen designed to identify essential residues in tRNA(Arg). The system consists of a tRNA(Arg) knockout strain and a set of vectors designed to rescue and select for variants of tRNA(Arg). By plasmid shuffling we selected inactive tRNA mutants that were further analyzed by northern blotting. The mutational analysis focused on the tRNA D and anticodon loops that contact the aminoacyl-tRNA synthetase. The anticodon triplet was excluded from the analysis because of its role in decoding the Arg codons. Most of the inactivating mutations are residues involved in tertiary interactions. These mutations had dramatic effects on tRNA(Arg) abundance. Other inactivating mutations were located in the anticodon loop, where they did not affect transcription and aminoacylation but probably altered interaction with the translation machinery. No lethal effects were observed when residues 16, 20 and 38 were individually mutated, despite the fact that they are involved in sequence-specific interactions with the aminoacyl-tRNA synthetase. However, the steady-state levels of the aminoacylated forms of U20A and U20G were decreased by a factor of 3.5-fold in vivo. This suggests that, unlike in the Escherichia coli tRNA(Arg):ArgRS system where residue 20 (A) is a major identity element, in yeast this position is of limited consequence. PMID- 12907714 TI - Identifying the methyltransferases for m(5)U747 and m(5)U1939 in 23S rRNA using MALDI mass spectrometry. AB - There are three sites of m(5)U modification in Escherichia coli stable RNAs: one at the invariant tRNA position U54 and two in 23S rRNA at the phylogenetically conserved positions U747 and U1939. Each of these sites is modified by its own methyltransferase, and the tRNA methyltransferase, TrmA, is well-characterised. Two open reading frames, YbjF and YgcA, are approximately 30% identical to TrmA, and here we determine the functions of these candidate methyltransferases using MALDI mass spectrometry. A purified recombinant version of YgcA retains its activity and specificity, and methylates U1939 in an RNA transcript in vitro. We were unable to generate a recombinant version of YbjF that retained in vitro activity, so the function of this enzyme was defined in vivo by engineering a ybjF knockout strain. Comparison of the methylation patterns in 23S rRNAs from YbjF(+) and YbjF(-) strains showed that the latter differed only in the lack of the m(5)U747 modification. With this report, the functions of all the E.coli m(5)U RNA methyltransferases are identified, and a more appropriate designation for YbjF would be RumB (RNA uridine methyltransferases B), in line with the recent nomenclature change for YgcA (now RumA). PMID- 12907715 TI - NMR study on the interaction between RPA and DNA decamer containing cis-syn cyclobutane pyrimidine dimer in the presence of XPA: implication for damage verification and strand-specific dual incision in nucleotide excision repair. AB - In mammalian cells, nucleotide excision repair (NER) is the major pathway for the removal of bulky DNA adducts. Many of the key NER proteins are members of the XP family (XPA, XPB, etc.), which was named on the basis of its association with the disorder xerodoma pigmentosum. Human replication protein A (RPA), the ubiquitous single-stranded DNA-binding protein, is another of the essential proteins for NER. RPA stimulates the interaction of XPA with damaged DNA by forming an RPA-XPA complex on damaged DNA sites. Binding of RPA to the undamaged DNA strand is most important during NER, because XPA, which directs the excision nucleases XPG and XPF, must bind to the damaged strand. In this study, nuclear magnetic resonance (NMR) spectroscopy was used to assess the binding of the tandem high affinity DNA binding domains, RPA-AB, and of the isolated domain RPA-A, to normal DNA and damaged DNA containing the cyclobutane pyrimidine dimer (CPD) lesion. Both RPA-A and RPA-AB were found to bind non- specifically to both strands of normal and CPD containing DNA duplexes. There were no differences observed when binding to normal DNA duplex was examined in the presence of the minimal DNA-binding domain of XPA (XPA-MBD). However, there is a drastic difference for CPD-damaged DNA duplex as both RPA-A and RPA-AB bind specifically to the undamaged strand. The strand-specific binding of RPA and XPA to the damaged duplex DNA shows that RPA and XPA play crucial roles in damage verification and guiding cleavage of damaged DNA during NER. PMID- 12907716 TI - RNA structure comparison, motif search and discovery using a reduced representation of RNA conformational space. AB - Given the wealth of new RNA structures and the growing list of RNA functions in biology, it is of great interest to understand the repertoire of RNA folding motifs. The ability to identify new and known motifs within novel RNA structures, to compare tertiary structures with one another and to quantify the characteristics of a given RNA motif are major goals in the field of RNA research; however, there are few systematic ways to address these issues. Using a novel approach for visualizing and mathematically describing macromolecular structures, we have developed a means to quantitatively describe RNA molecules in order to rapidly analyze, compare and explore their features. This approach builds on the alternative eta,theta convention for describing RNA torsion angles and is executed using a new program called PRIMOS. Applying this methodology, we have successfully identified major regions of conformational change in the 50S and 30S ribosomal subunits, we have developed a means to search the database of RNA structures for the prevalence of known motifs and we have classified and identified new motifs. These applications illustrate the powerful capabilities of our new RNA structural convention, and they suggest future adaptations with important implications for bioinformatics and structural genomics. PMID- 12907717 TI - Molecular handcuffing of the relaxosome at the origin of conjugative transfer of the plasmid R1162. AB - The assembly of plasmid-encoded proteins at a unique site (oriT) on the plasmid R1162, to form a complex called the relaxosome, is required for conjugative transfer of the plasmid and for negative regulation of neighboring promoters. Two dimensional chloroquine gel electrophoresis was used to show that oriTs are physically coupled at the relaxosome. This interaction requires all the relaxosome proteins, which are assembled into a structure resulting in a decrease in the average linking number of the plasmid DNA in the cell. Molecules with higher superhelical densities are preferentially selected for assembly of the relaxosome. Genetic data obtained earlier indicate that the molecular coupling reported here is a 'handcuffing' reaction that contributes to the regulation of adjacent plasmid promoters. However, although these promoters affect the expression of the genes for replication, plasmid copy-control is regulated independently. This is the first time 'handcuffing' has been observed at an oriT, and its possible significance for transfer is discussed. PMID- 12907718 TI - Efficient incorporation of positively charged 2', 3'-dideoxynucleoside-5' triphosphates by DNA polymerases and their application in 'direct-load' DNA sequencing. AB - A series of charge-modified, dye-labeled 2', 3'-dideoxynucleoside-5' triphosphates have been synthesized and evaluated as reagents for dye-terminator DNA sequencing. Unlike the commonly used dye-labeled terminators, these terminators possess a net positive charge and migrate in the opposite direction to dye-labeled Sanger fragments during electrophoresis. Post-sequencing reaction purification is not required to remove unreacted nucleotide or associated breakdown products prior to electrophoresis. Thus, DNA sequencing reaction mixtures can be loaded directly onto a separating medium such as a sequencing gel. The charge-modified nucleotides have also been shown to be more efficiently incorporated by a number of DNA polymerases than regular dye-labeled dideoxynucleotide terminators or indeed normal dideoxynucleoside-5' triphosphates. PMID- 12907719 TI - Cell cycle arrest and apoptosis provoked by UV radiation-induced DNA damage are transcriptionally highly divergent responses. AB - DNA damage caused by UV radiation initiates cellular recovery mechanisms, which involve activation of DNA damage response pathways, cell cycle arrest and apoptosis. To assess cellular transcriptional responses to UVC-induced DNA damage we compared time course responses of human skin fibroblasts to low and high doses of UVC radiation known to induce a transient cellular replicative arrest or apoptosis, respectively. UVC radiation elicited >3-fold changes in 460 out of 12,000 transcripts and 89% of these represented downregulated transcripts. Only 5% of the regulated genes were common to both low and high doses of radiation. Cells inflicted with a low dose of UVC exhibited transcription profiles demonstrating transient regulation followed by recovery, whereas the responses were persistent after the high dose. A detailed clustering analysis and functional classification of the targets implied regulation of biologically divergent responses and suggested involvement of transcriptional and translational machinery, inflammatory, anti-proliferative and anti-angiogenic responses. The data support the notion that UVC radiation induces prominent, dose dependent downregulation of transcription. However, the data strongly suggest that transcriptional repression is also target gene selective. Furthermore, the results demonstrate that dose-dependent induction of cell cycle arrest and apoptosis by UVC radiation are transcriptionally highly distinct responses. PMID- 12907720 TI - Two protein-protein interaction sites on the spliceosome-associated human cyclophilin CypH. AB - Cyclophilins are a family of proteins that share a common, highly conserved sequence motif. Cyclophilins bind transiently to other proteins and facilitate their folding. One member of the family, hCypH, is part of the human spliceosomal [U4/U6.U5] tri-snRNP complex; it associates specifically and stably with the U4/U6-specific protein 60K. Here, we demonstrate that recombinant hCypH exhibits peptidyl-prolyl isomerase (PPIase) activity, and describe mutagenesis studies demonstrating that it shares the catalytic pocket with other members of the cyclophilin family. However, neither the PPIase activity nor the catalytic pocket is required for binding of protein 60K. Rather, hCypH contains a small insertion in a loop of the otherwise conserved cyclophilin backbone, and this minor change creates a highly specific binding site that is responsible for the association of this cyclophilin, but not others, with protein 60K. hCypH is thus the first small cyclophilin shown to have a second protein-protein interaction site and the ability to bind stably to another protein. Since the catalytic pocket and the second binding site are located on opposite sides of the cyclophilin structure, this opens up the interesting possibility that hCypH may serve as a bridge mediating interactions between protein 60K of the U4/U6 snRNP and other as yet unknown factors. PMID- 12907721 TI - Methylation of CpG dinucleotides in the open reading frame of a testicular germ cell-specific intronless gene, Tact1/Actl7b, represses its expression in somatic cells. AB - Methylation of CpG islands spanning promoter regions is associated with control of gene expression. However, it is considered that methylation of exonic CpG islands without promoter is not related to gene expression, because such exonic CpG islands are usually distant from the promoter. Whether methylation of exonic CpG islands near the promoter, as in the case of a CpG-rich intronless gene, causes repression of the promoter remains unknown. To gain insight into this issue, we investigated the distribution and methylation status of CpG dinucleotides in the mouse Tact1/Actl7b gene, which is intronless and expressed exclusively in testicular germ cells. The region upstream to the gene was poor in CpG, with CpG dinucleotides absent from the core promoter. However, a CpG island was found inside the open reading frame (ORF). Analysis of the methylation status of the Tact1/Actl7b gene including the 5'-flanking area demonstrated that all CpG sites were methylated in somatic cells, whereas these sites were unmethylated in the Tact1/Actl7b-positive testis. Trans fection experiments with in vitro methylated constructs indicated that methylation of the ORF but not 5' upstream repressed Tact1/Actl7b promoter activity in somatic cells. Similar effects of ORF methylation on the promoter activity were observed in testicular germ cells. These are the first results indicating that methylation of the CpG island in the ORF represses its promoter in somatic cells and demethylation is necessary for gene expression in spermatogenic cells. PMID- 12907722 TI - Apoptin protein multimers form distinct higher-order nucleoprotein complexes with DNA. AB - The chicken anaemia virus-derived protein apoptin is a tumour-specific cell killing agent. It is biologically active as a highly stable, multimeric complex, consisting of 30-40 monomers. In tumour cells, but negligibly in normal cells, apoptin is imported into the nucleus prior to the induction of apoptosis. Immunoelectron microscopic data we report here indicate that apoptin predominantly co-localises with heterochromatin and nucleoli within tumour cells. Apoptin's preference for these DNA-dense nuclear bodies may be explained by our finding that apoptin cooperatively forms distinct superstructures with DNA in vitro. These superstructures do not grow beyond a diameter of approximately 200 nm, containing up to 20 multimeric apoptin complexes and approximately 3 kb of DNA. Furthermore, we show a single apoptin multimer to have eight independent, non-specific DNA-binding sites which preferentially bind strand ends, but which can also collaborate to bind longer stretches of DNA. Apoptin's high affinity for naked, undecorated double- and single-stranded DNA and for DNA fibre ends suggests that it may also capture such DNA in superstructures in vivo. Since these forms of DNA are predominantly found in transcriptionally active, replicating and damaged DNA, apoptin could be triggering apoptosis by interfering with DNA transcription and synthesis. PMID- 12907723 TI - Structures of Escherichia coli DNA mismatch repair enzyme MutS in complex with different mismatches: a common recognition mode for diverse substrates. AB - We have refined a series of isomorphous crystal structures of the Escherichia coli DNA mismatch repair enzyme MutS in complex with G:T, A:A, C:A and G:G mismatches and also with a single unpaired thymidine. In all these structures, the DNA is kinked by approximately 60 degrees upon protein binding. Two residues widely conserved in the MutS family are involved in mismatch recognition. The phenylalanine, Phe 36, is seen stacking on one of the mismatched bases. The same base is also seen forming a hydrogen bond to the glutamate Glu 38. This hydrogen bond involves the N7 if the base stacking on Phe 36 is a purine and the N3 if it is a pyrimidine (thymine). Thus, MutS uses a common binding mode to recognize a wide range of mismatches. PMID- 12907724 TI - The bacterial transposon Tn7 causes premature polyadenylation of mRNA in eukaryotic organisms: TAGKO mutagenesis in filamentous fungi. AB - TAGKO is a Tn7-based transposition system for genome wide mutagenesis in filamentous fungi. The effects of transposon insertion on the expression of TAGKO alleles were examined in Magnaporthe grisea and Mycosphaerella graminicola. Northern analysis showed that stable, truncated transcripts were expressed in the TAGKO mutants. Mapping of the 3'-ends of TAGKO cDNAs revealed that they all contain Tn7 end sequences, regardless of the transposon orientation. Polyadenylation signals characteristic of eukaryotic genes, preceded by stop codons in all frames, are located in both ends of the bacterial transposon. Thus, TAGKO transcripts are prematurely polyadenylated, and truncated proteins are predicted to be translated in the fungal mutants. Depending on the extent of protein truncation, TAGKO mutations in HPD4 (encoding p-hydroxyphenylpyruvate dioxygenase) resulted in tyrosine sensitivity in the two fungi. Similarly, a particular M.grisea CBS1 (encoding cystathionine beta-synthase) TAGKO cDNA failed to complement cysteine auxotrophy in a yeast CBS mutant. TAGKO, therefore, represents a useful tool for in vivo study of truncated gene products in filamentous fungi. PMID- 12907725 TI - Large scale study of protein domain distribution in the context of alternative splicing. AB - Alternative splicing plays an important role in processes such as development, differentiation and cancer. With the recent increase in the estimates of the number of human genes that undergo alternative splicing from 5 to 35-59%, it is becoming critical to develop a better understanding of its functional consequences and regulatory mechanisms. We conducted a large scale study of the distribution of protein domains in a curated data set of several thousand genes and identified protein domains disproportionately distributed among alternatively spliced genes. We also identified a number of protein domains that tend to be spliced out. Both the proteins having the disproportionately distributed domains as well as those with spliced-out domains are predominantly involved in the processes of cell communication, signaling, development and apoptosis. These proteins function mostly as enzymes, signal transducers and receptors. Somewhat surprisingly, 28% of all occurrences of spliced-out domains are not effected by straightforward exclusion of exons coding for the domains but by inclusion or exclusion of other exons to shift the reading frame while retaining the exons coding for the domains in the final transcripts. PMID- 12907726 TI - Alterations in the intracellular level of a protein subunit of human RNase P affect processing of tRNA precursors. AB - The human ribonucleoprotein ribonuclease P (RNase P), processing tRNA, has at least 10 distinct protein subunits. Many of these subunits, including the autoimmune antigen Rpp38, are shared by RNase MRP, a ribonucleoprotein enzyme required for processing of rRNA. We here show that constitutive expression of exogenous, tagged Rpp38 protein in HeLa cells affects processing of tRNA precursors. Alterations in the site-specific cleavage and in the steady-state level of 3' sequences of the internal transcribed spacer 1 of rRNA are also observed. These processing defects are accompanied by selective shut-off of expression of Rpp38 and by low expression of the tagged protein. RNase P purified from these cells exhibits impaired activity in vitro. Moreover, inhibition of Rpp38 by the use of small interfering RNA causes accumulation of the initiator methionine tRNA precursor. Expression of other protein components, but not of the H1 RNA subunit, is coordinately inhibited. Our results reveal that normal expression of Rpp38 is required for the biosynthesis of intact RNase P and for the normal processing of stable RNA in human cells. PMID- 12907727 TI - Zinc finger-dependent HIV-1 nucleocapsid protein-TAR RNA interactions. AB - In the minus-strand transfer step of HIV-1 reverse transcription, the nucleocapsid protein (NC) promotes annealing of the 3' 'R' (repeat) region of the RNA genome to its complementary sequence located in the newly synthesized minus strand strong-stop DNA. The R region contains the highly stable transactivation response (TAR) RNA hairpin. To gain insights into the molecular details of TAR RNA-NC interactions, we carried out hydroxyl radical footprinting, as well as gel shift and fluorescence anisotropy binding assays using wild-type and mutant forms of NC. Our results support the conclusion that NC variants with mutations in their zinc finger domains have dramatically altered TAR RNA binding interactions relative to wild-type NC. These data demonstrate that a specific zinc finger architecture is required for optimal TAR RNA binding, and help to explain the requirement for the zinc finger motifs of NC in its role as a nucleic acid chaperone in minus-strand transfer. PMID- 12907728 TI - The sequence and analysis of Trypanosoma brucei chromosome II. AB - We report here the sequence of chromosome II from Trypanosoma brucei, the causative agent of African sleeping sickness. The 1.2-Mb pairs encode about 470 predicted genes organised in 17 directional clusters on either strand, the largest cluster of which has 92 genes lined up over a 284-kb region. An analysis of the GC skew reveals strand compositional asymmetries that coincide with the distribution of protein-coding genes, suggesting these asymmetries may be the result of transcription-coupled repair on coding versus non-coding strand. A 5-cM genetic map of the chromosome reveals recombinational 'hot' and 'cold' regions, the latter of which is predicted to include the putative centromere. One end of the chromosome consists of a 250-kb region almost exclusively composed of RHS (pseudo)genes that belong to a newly characterised multigene family containing a hot spot of insertion for retroelements. Interspersed with the RHS genes are a few copies of truncated RNA polymerase pseudogenes as well as expression site associated (pseudo)genes (ESAGs) 3 and 4, and 76 bp repeats. These features are reminiscent of a vestigial variant surface glycoprotein (VSG) gene expression site. The other end of the chromosome contains a 30-kb array of VSG genes, the majority of which are pseudogenes, suggesting that this region may be a site for modular de novo construction of VSG gene diversity during transposition/gene conversion events. PMID- 12907730 TI - ADAR2 A-->I editing: site selectivity and editing efficiency are separate events. AB - ADAR enzymes, adenosine deaminases that act on RNA, form a family of RNA editing enzymes that convert adenosine to inosine within RNA that is completely or largely double-stranded. Site-selective A-->I editing has been detected at specific sites within a few structured pre-mRNAs of metazoans. We have analyzed the editing selectivity of ADAR enzymes and have chosen to study the naturally edited R/G site in the pre-mRNA of the glutamate receptor subunit B (GluR-B). A comparison of editing by ADAR1 and ADAR2 revealed differences in the specificity of editing. Our results show that ADAR2 selectively edits the R/G site, while ADAR1 edits more promiscuously at several other adenosines in the double-stranded stem. To further understand the mechanism of selective ADAR2 editing we have investigated the importance of internal loops in the RNA substrate. We have found that the immediate structure surrounding the editing site is important. A purine opposite to the editing site has a negative effect on both selectivity and efficiency of editing. More distant internal loops in the substrate were found to have minor effects on site selectivity, while efficiency of editing was found to be influenced. Finally, changes in the RNA structure that affected editing did not alter the binding abilities of ADAR2. Overall these findings suggest that binding and catalysis are independent events. PMID- 12907729 TI - The DNA sequence of chromosome I of an African trypanosome: gene content, chromosome organisation, recombination and polymorphism. AB - The African trypanosome, Trypanosoma brucei, causes sleeping sickness in humans in sub-Saharan Africa. Here we report the sequence and analysis of the 1.1 Mb chromosome I, which encodes approximately 400 predicted genes organised into directional clusters, of which more than 100 are located in the largest cluster of 250 kb. A 160-kb region consists primarily of three gene families of unknown function, one of which contains a hotspot for retroelement insertion. We also identify five novel gene families. Indeed, almost 20% of predicted genes are members of families. In some cases, tandemly arrayed genes are 99-100% identical, suggesting an active process of amplification and gene conversion. One end of the chromosome consists of a putative bloodstream-form variant surface glycoprotein (VSG) gene expression site that appears truncated and degenerate. The other chromosome end carries VSG and expression site-associated genes and pseudogenes over 50 kb of subtelomeric sequence where, unusually, the telomere-proximal VSG gene is oriented away from the telomere. Our analysis includes the cataloguing of minor genetic variations between the chromosome I homologues and an estimate of crossing-over frequency during genetic exchange. Genetic polymorphisms are exceptionally rare in sequences located within and around the strand-switches between several gene clusters. PMID- 12907731 TI - Role of cysteine amino acid residues on the RNA binding activity of human thymidylate synthase. AB - The role of cysteine sulfhydryl residues on the RNA binding activity of human thymidylate synthase (TS) was investigated by mutating each cysteine residue on human TS to a corresponding alanine residue. Enzymatic activities of TS:C43A and TS:C210A mutant proteins were nearly identical to wild-type TS, while TS:C180A and TS:C199A mutants expressed >80% of wild-type enzyme activity. In contrast, TS:C195A was completely inactive. Mutant proteins, TS:C195A, TS:C199A and TS:C210A, retained RNA binding activity to nearly the same degree as wild-type human TS. RNA binding activity of TS:C43A was reduced by 30% when compared to wild-type TS, while TS:C180A was completely devoid of RNA binding activity. In vitro translation studies confirmed that mutant proteins TS:C43A, TS:C195A, TS:C199A and TS:C210A, significantly repressed human TS mRNA translation, while TS:C180A was unable to do so. To confirm the in vivo significance of the cysteine sulfhydryl residue, mutant proteins TS:C180A and TS:C195A were each expressed in human colon cancer HCT-C18:TS(-) cells that expressed a functionally inactive TS. A recombinant luciferase reporter gene under the control of a TS-response element was co-transfected into these same cells, and luciferase activity increased in the presence of the TS:C195A mutant TS protein to a level similar to that observed upon expression of wild-type TS protein. In contrast, luciferase activity remained unchanged in cells expressing the TS:C180A mutant protein. Taken together, these findings identify Cys-180 as a critical residue for the in vitro and in vivo translational regulatory effects of human TS. PMID- 12907732 TI - Biochemical analysis of components of the pre-replication complex of Archaeoglobus fulgidus. AB - The eukaryotic pre-replication complex is assembled at replication origins in a reaction called licensing. Licensing involves the interactions of a variety of proteins including the origin recognition complex (ORC), Cdc6 and the Mcm2-7 helicase, homologues of which are also found in archaea. The euryarchaeote Archaeoglobus fulgidus encodes two genes with homology to Orc/Cdc6 and a single Mcm homologue. The A.fulgidus Mcm protein and one Orc/Cdc6 homologue have been purified and investigated in vitro. The Mcm protein is an ATP-dependent, hexameric helicase that can unwind between 200 and 400 bp of duplex DNA. Deletion of 112 amino acids from the N-terminus of A.f Mcm produced a protein, which was still capable of forming a hexamer, was competent in DNA binding and was able to unwind at least 1 kb of duplex DNA. The purified Orc/Cdc6 homologue was also able to bind DNA. Both Mcm and Orc/Cdc6 show a preference for specific DNA structures, namely molecules containing a single stranded bubble that mimics early replication intermediates. Nuclease protection showed that the binding sites for Mcm and Orc/Cdc6 overlap. The Orc/Cdc6 protein bound more tightly to these substrates and was able to displace pre-bound Mcm hexamer. PMID- 12907734 TI - Single nucleotide polymorphisms (SNPs) that map to gaps in the human SNP map. AB - An international effort is underway to generate a comprehensive haplotype map (HapMap) of the human genome represented by an estimated 300,000 to 1 million 'tag' single nucleotide polymorphisms (SNPs). Our analysis indicates that the current human SNP map is not sufficiently dense to support the HapMap project. For example, 24.6% of the genome currently lacks SNPs at the minimal density and spacing that would be required to construct even a conservative tag SNP map containing 300,000 SNPs. In an effort to improve the human SNP map, we identified 140,696 additional SNP candidates using a new bioinformatics pipeline. Over 51,000 of these SNPs mapped to the largest gaps in the human SNP map, leading to significant improvements in these regions. Our SNPs will be immediately useful for the HapMap project, and will allow for the inclusion of many additional genomic intervals in the final HapMap. Nevertheless, our results also indicate that additional SNP discovery projects will be required both to define the haplotype architecture of the human genome and to construct comprehensive tag SNP maps that will be useful for genetic linkage studies in humans. PMID- 12907733 TI - Sequence-structure-function relationships of Tgs1, the yeast snRNA/snoRNA cap hypermethylase. AB - The Saccharomyces cerevisiae Tgs1 methyltransferase (MTase) is responsible for conversion of the m(7)G caps of snRNAs and snoRNAs to a 2,2,7- trimethylguanosine structure. To learn more about the evolutionary origin of Tgs1 and to identify structural features required for its activity, we performed a structure-function study. By using sequence comparison and phylogenetic analysis, we found that Tgs1 shows strongest similarity to Mj0882, a protein related to a family comprised of bacterial rRNA:m(2)G MTases RsmC and RsmD. The structural information of Mj0882 was used to build a homology model of Tgs1p which allowed us to predict the range of the minimal globular MTase domain and the localization of other residues that may be important for enzyme function. To further characterize functional domains of Tgs1, mutants were constructed and tested for their effects on cell viability, subcellular localization and binding to the small nuclear ribonucleoproteins (snRNPs) and small nucleolar RNPs (snoRNPs). We found that the N-terminal domain of the hypermethylase is dispensable for binding to the common snRNPs and snoRNPs proteins but essential for correct nucleolar localization. Site- directed mutagenesis of Tgs1 allowed also the identification of the residues likely to be involved in the formation of the m7G-binding site and the catalytic center. PMID- 12907735 TI - Cloning, functional analysis and post-transcriptional regulation of a type II DNA topoisomerase from Leishmania infantum. A new potential target for anti-parasite drugs. AB - We identified a type II topoisomerase enzyme from Leishmania infantum, a parasite protozoon causing disease in humans. This protein, named Li topo II, which displays a variable C-terminal end, is located in the kinetoplast. The cloned gene encoding Li-TOP2 compensates for the slow growth of topo II-deficient mutants of Saccharomyces cerevisiae, resulting in a catalytically active DNA topoisomerase in yeast. Analysis of the specific mRNA levels of the Li-TOP2 gene showed variations throughout the parasite cell cycle in synchronized cells as well as between the distinct forms of the parasite. Thus, the enzyme had higher levels of mRNA expression in the highly infective intracellular form of the parasite, the amastigote, than in the extracellular promastigote form, suggesting a relation with the distinct developmental and infectious phases of the protozoon. In addition, western blot analysis showed differences in protein expression between the proliferative and non-proliferative forms of L.infantum promastigotes, which displayed similar levels of mRNA. This indicated possible post-transcriptional regulation mechanisms. The data suggest that Li topo II has a part in DNA decatenation and probably at the initial stages of proliferation in the intracellular form of L.infantum, a parasite that has to proliferate into the host macrophage to survive its hostile environment in its first moments of intracellular infection. PMID- 12907736 TI - An important role for RUNX3 in human L1 transcription and retrotransposition. AB - LINE-1s (long interspersed nuclear elements-1) are abundant non-LTR retrotransposons that comprise 17% of the human genome. The 5' untranslated region (5'UTR) of human L1 (L1Hs) houses a poorly understood internal promoter. Here we report that mutations at a putative runt-domain transcription factor (RUNX) site (+83 to +101) in the 5'UTR decreased L1Hs transcription and retrotransposition in cell culture-based assays. Exogenous expression of RUNX3, but not the other two RUNX family members, RUNX1 and RUNX2, increased L1Hs transcription and retrotransposition, which were otherwise decreased by siRNAs targeting RUNX3 and a dominant negative RUNX. Further more, the specific interaction between RUNX3 and its binding site was demonstrated by an electrophoretic mobility shift assay (EMSA) using an anti-RUNX3 antibody. Interestingly, RUNX3 may also regulate the antisense promoter activity of L1Hs 5'UTR via another putative RUNX site (+526 to +508), as revealed by site-directed mutations and exogenous expression of RUNX factors. Our results indicate an important role for RUNX3 in L1Hs retrotransposition as well as transcription from its 5'UTR in both sense and antisense directions, and they should contribute to our understanding of the mechanism underlying L1Hs retrotransposition and its impact on the expression of adjacent cellular genes. PMID- 12907738 TI - Characterisation of the substrate specificity of homogeneous vaccinia virus uracil-DNA glycosylase. AB - The decision to stop smallpox vaccination and the loss of specific immunity in a large proportion of the population could jeopardise world health due to the possibility of a natural or provoked re-emergence of smallpox. Therefore, it is mandatory to improve the current capability to prevent or treat such infections. The DNA repair protein uracil-DNA glycosylase (UNG) is one of the viral enzymes important for poxvirus pathogenesis. Consequently, the inhibition of UNG could be a rational strategy for the treatment of infections with poxviruses. In order to develop inhibitor assays for UNG, as a first step, we have characterised the recombinant vaccinia virus UNG (vUNG) and compared it with the human nuclear form (hUNG2) and catalytic fragment (hUNG) UNG. In contrast to hUNG2, vUNG is strongly inhibited in the presence of 7.5 mM MgCl(2). We have shown that highly purified vUNG is not inhibited by a specific uracil-DNA glycosylase inhibitor. Interestingly, both viral and human enzymes preferentially excise uracil when it is opposite to cytosine. The present study provides the basis for the design of specific inhibitors for vUNG. PMID- 12907737 TI - Alanine-scanning mutagenesis of the predicted rRNA-binding domain of ErmC' redefines the substrate-binding site and suggests a model for protein-RNA interactions. AB - The Erm family of adenine-N(6) methyltransferases (MTases) is responsible for the development of resistance to macrolide-lincosamide-streptogramin B antibiotics through the methylation of 23S ribosomal RNA. Hence, these proteins are important potential drug targets. Despite the availability of the NMR and crystal structures of two members of the family (ErmAM and ErmC', respectively) and extensive studies on the RNA substrate, the substrate-binding site and the amino acids involved in RNA recognition by the Erm MTases remain unknown. It has been proposed that the small C-terminal domain functions as a target-binding module, but this prediction has not been tested experimentally. We have undertaken structure-based mutational analysis of 13 charged or polar residues located on the predicted rRNA-binding surface of ErmC' with the aim to identify the area of protein-RNA interactions. The results of in vivo and in vitro analyses of mutant protein suggest that the key RNA-binding residues are located not in the small domain, but in the large catalytic domain, facing the cleft between the two domains. Based on the mutagenesis data, a preliminary three-dimensional model of ErmC' complexed with the minimal substrate was constructed. The identification of the RNA-binding site of ErmC' may be useful for structure-based design of novel drugs that do not necessarily bind to the cofactor-binding site common to many S adenosyl-L- methionine-dependent MTases, but specifically block the substrate binding site of MTases from the Erm family. PMID- 12907739 TI - Impact of surface chemistry and blocking strategies on DNA microarrays. AB - The surfaces and immobilization chemistries of DNA microarrays are the foundation for high quality gene expression data. Four surface modification chemistries, poly-L-lysine (PLL), 3-glycidoxypropyltrimethoxysilane (GPS), DAB-AM poly(propyleminime hexadecaamine) dendrimer (DAB) and 3 aminopropyltrimethoxysilane (APS), were evaluated using cDNA and oligonucleotide sub-arrays. Two un-silanized glass surfaces, RCA-cleaned and immersed in Tris EDTA buffer were also studied. DNA on amine-modified surfaces was fixed by UV (90 mJ/cm(2)), while DNA on GPS-modified surfaces was immobilized by covalent coupling. Arrays were blocked with either succinic anhydride (SA), bovine serum albumin (BSA) or left unblocked prior to hybridization with labeled PCR product. Quality factors evaluated were surface affinity for cDNA versus oligonucleotides, spot and background intensity, spotting concentration and blocking chemistry. Contact angle measurements and atomic force microscopy were preformed to characterize surface wettability and morphology. The GPS surface exhibited the lowest background intensity regardless of blocking method. Blocking the arrays did not affect raw spot intensity, but affected background intensity on amine surfaces, BSA blocking being the lowest. Oligonucleotides and cDNA on unblocked GPS-modified slides gave the best signal (spot-to-background intensity ratio). Under the conditions evaluated, the unblocked GPS surface along with amine covalent coupling was the most appropriate for both cDNA and oligonucleotide microarrays. PMID- 12907740 TI - Dendrimeric coating of glass slides for sensitive DNA microarrays analysis. AB - Successful use and reliability of microarray technology is highly dependent on several factors, including surface chemistry parameters and accessibility of cDNA targets to the DNA probes fixed onto the surface. Here, we show that functionalisation of glass slides with homemade dendrimers allow production of more sensitive and reliable DNA microarrays. The dendrimers are nanometric structures of size-controlled diameter with aldehyde function at their periphery. Covalent attachment of these spherical reactive chemical structures on amino silanised glass slides generates a reactive approximately 100 A layer onto which amino-modified DNA probes are covalently bound. This new grafting chemistry leads to the formation of uniform and homogenous spots. More over, probe concentration before spotting could be reduced from 0.2 to 0.02 mg/ml with PCR products and from 20 to 5 micro M with 70mer oligonucleotides without affecting signal intensities after hybridisation with Cy3- and Cy5-labelled targets. More interestingly, while the binding capacity of captured probes on dendrimer activated glass surface (named dendrislides) is roughly similar to other functionalised glass slides from commercial sources, detection sensitivity was 2 fold higher than with other available DNA microarrays. This detection limit was estimated to 0.1 pM of cDNA targets. Altogether, these features make dendrimer activated slides ideal for manufacturing cost-effective DNA arrays applicable for gene expression and detection of mutations. PMID- 12907742 TI - Combination of DNA-directed immobilization and immuno-PCR: very sensitive antigen detection by means of self-assembled DNA-protein conjugates. AB - An assay for very sensitive antigen detection is described which takes advantage of the self- assembly capabilities of semi-synthetic conjugates of DNA and proteins. The general scheme of this assay is similar to a two-sided (sandwich) enzyme-linked immunoassay (ELISA); however, covalent single-stranded DNA streptavidin (STV) conjugates, capable of hybridizing to complementary surface bound DNA oligomers, are utilized for the effective immobilization of either capture antibodies or antigens, rather than the chemi- or physisorption usually applied in ELISA. Immuno-PCR (IPCR) is employed as a method for signal generation, utilizing oligomeric reagents obtained by self-assembly of STV, biotinylated DNA and antibodies. In three different model systems, detecting human IgG, rabbit IgG or carcinoembryonic antigen, this combination allowed one to increase the sensitivity of the analogous ELISA approximately 1000-fold. For example, <0.1 amol/ micro l (15 pg/ml) of rabbit IgG was detectable. The immunoassay can be carried out in a single step by tagging the analyte with both reagents for capture and read-out simultaneously, thereby significantly reducing handling time and costs of analysis. Moreover, as the spatial selectivity of target immobilization is determined by the specificity of DNA base pairing, the assay is particularly suited for miniaturized microfluidics and lab-on-a-chip devices. PMID- 12907743 TI - Method to protect a targeted amino acid residue during random mutagenesis. AB - To generate a random mutant library that is free from mutation at a particular amino acid residue, we replace the codon of interest with a detachable, short DNA sequence containing a BsaXI recognition site. After PCR mutagenesis, this sequence is removed and intramolecular ligation of the sequences flanking the insert regenerates the gene. The three-base cohesive ends for ligation correspond to the codon for the targeted residue and any sequences with mutations at this site will fail to ligate. As a result, only the variants that are free from mutation at this site are in the proper reading frame. In a random library of C(30) carotenoid synthase CrtM, this method was used to exclude readily accessible mutations at position F26, which confer C(40) synthase function. This enabled us to identify two additional mutations, W38C and E180G, which confer the same phenotype but are present in the random library at much lower frequencies. PMID- 12907741 TI - Gene expression analysis using single molecule detection. AB - Recent developments of single molecule detection techniques and in particular the introduction of fluorescence correlation spectroscopy (FCS) led to a number of important applications in biological research. We present a unique approach for the gene expression analysis using dual-color cross-correlation. The expression assay is based on gene-specific hybridization of two dye-labeled DNA probes to a selected target gene. The counting of the dual-labeled molecules within the solution allows the quantification of the expressed gene copies in absolute numbers. As detection and analysis by FCS can be performed at the level of single molecules, there is no need for any type of amplification. We describe the gene expression assay and present data demonstrating the capacity of this novel technology. In order to prove the gene specificity, we performed experiments with gene-depleted total cDNA. The biological application was demonstrated by quantifying selected high, medium and low abundant genes in cDNA prepared from HL 60 cells. PMID- 12907744 TI - Detection of methylation of human p16(Ink4a) gene 5'-CpG islands by electrochemical method coupled with linker-PCR. AB - Aberrant DNA methylation of the CpG site is among the earliest and most frequent alterations in cancer. Detection of promoter hypermethylation of cancer-related genes may be useful for cancer diagnosis or the detection of recurrence. p16, an inhibitor of the cyclin D-dependent protein kinases, is a classical tumor suppressor gene, and its inactivation is closely associated with carcinogenesis. p16 hypermethylation could be detected in each stage, which is consistent with the finding that aberrant methylation of p16 is a very early event in carcinogenesis. We have developed an electrochemical procedure for detecting DNA methylation of the human p16(Ink4a) gene. The procedure is based on the coupling of DNA electrochemical sensors with linker-PCR- amplified DNA from human gastric tumor tissue and whole blood cells of healthy human. The synthesized oligonucleotide was immobilized on the modified gold electrode to fabricate a DNA biosensor. The hybridization reaction on the electrode surface was monitored by cyclic voltammogram (CV) and square wave voltammogram (SWV), using [Co(phen)(3)](ClO(4))(3) as a redox indicator. Methylation status of human p16(Ink4a) gene was detected and the results were validated by bisulfite DNA sequencing. A good reproducibility was observed in several parallel experiments. The coupling of DNA electrochemical sensors with PCR allowed quick detection and have the potential of the quantitative evaluation of the methylation status of the human p16(Ink4a) gene. PMID- 12907745 TI - Mathematics of quantitative kinetic PCR and the application of standard curves. AB - Fluorescent monitoring of DNA amplification is the basis of real-time PCR, from which target DNA concentration can be determined from the fractional cycle at which a threshold amount of amplicon DNA is produced. Absolute quantification can be achieved using a standard curve constructed by amplifying known amounts of target DNA. In this study, the mathematics of quantitative PCR are examined in detail, from which several fundamental aspects of the threshold method and the application of standard curves are illustrated. The construction of five replicate standard curves for two pairs of nested primers was used to examine the reproducibility and degree of quantitative variation using SYBER Green I fluorescence. Based upon this analysis the application of a single, well- constructed standard curve could provide an estimated precision of +/-6-21%, depending on the number of cycles required to reach threshold. A simplified method for absolute quantification is also proposed, in which quantitative scale is determined by DNA mass at threshold. PMID- 12907746 TI - A sensitive transcriptome analysis method that can detect unknown transcripts. AB - We have developed an AFLP-based gene expression profiling method called 'high coverage expression profiling' (HiCEP) analysis. By making improvements to the selective PCR technique we have reduced the rate of false positive peaks to approximately 4% and consequently the number of peaks, including overlapping peaks, has been markedly decreased. As a result we can determine the relationship between peaks and original transcripts unequivocally. This will make it practical to prepare a database of all peaks, allowing gene assignment without having to isolate individual peaks. This precise selection also enables us to easily clone peaks of interest and predict the corresponding gene for each peak in some species. The procedure is highly reproducible and sensitive enough to detect even a 1.2-fold difference in gene expression. Most importantly, the low false positive rate enables us to analyze gene expression with wide coverage by means of four instead of six nucleotide recognition site restriction enzymes for fingerprinting mRNAs. Therefore, the method detects 70-80% of all transcripts, including non-coding transcripts, unknown and known genes. Moreover, the method requires no sequence information and so is applicable even to eukaryotes for which there is no genome information available. PMID- 12907748 TI - A model-based analysis of microarray experimental error and normalisation. AB - A statistical model is proposed for the analysis of errors in microarray experiments and is employed in the analysis and development of a combined normalisation regime. Through analysis of the model and two-dye microarray data sets, this study found the following. The systematic error introduced by microarray experiments mainly involves spot intensity-dependent, feature-specific and spot position-dependent contributions. It is difficult to remove all these errors effectively without a suitable combined normalisation operation. Adaptive normalisation using a suitable regression technique is more effective in removing spot intensity-related dye bias than self-normalisation, while regional normalisation (block normalisation) is an effective way to correct spot position dependent errors. However, dye-flip replicates are necessary to remove feature specific errors, and also allow the analyst to identify the experimentally introduced dye bias contained in non-self-self data sets. In this case, the bias present in the data sets may include both experimentally introduced dye bias and the biological difference between two samples. Self-normalisation is capable of removing dye bias without identifying the nature of that bias. The performance of adaptive normalisation, on the other hand, depends on its ability to correctly identify the dye bias. If adaptive normalisation is combined with an effective dye bias identification method then there is no systematic difference between the outcomes of the two methods. PMID- 12907747 TI - Restriction site tagged (RST) microarrays: a novel technique to study the species composition of complex microbial systems. AB - We have developed a new type of microarray, restriction site tagged (RST), for example NotI, microarrays. In this approach only sequences surrounding specific restriction sites (i.e. NotI linking clones) were used for generating microarrays. DNA was labeled using a new procedure, NotI representation, where only sequences surrounding NotI sites were labeled. Due to these modifications, the sensitivity of RST microarrays increases several hundred-fold compared to that of ordinary genomic microarrays. In a pilot experiment we have produced NotI microarrays from Gram-positive and Gram-negative bacteria and have shown that even closely related Escherichia coli strains can be easily discriminated using this technique. For example, two E.coli strains, K12 and R2, differ by less than 0.1% in their 16S rRNA sequences and thus the 16S rRNA sequence would not easily discriminate between these strains. However, these strains showed distinctly different hybridization patterns with NotI microarrays. The same technique can be adapted to other restriction enzymes as well. This type of microarray opens the possibility not only for studies of the normal flora of the gut but also for any problem where quantitative and qualitative analysis of microbial (or large viral) genomes is needed. PMID- 12907750 TI - A chemical enucleation method for the transfer of mitochondrial DNA to rho(o) cells. AB - The study of pathogenic mitochondrial DNA mutations has, in most cases, relied on the production of transmitochondrial cybrids. Although the procedure to produce such cybrids is well established, it is laborious and cumbersome. Moreover, the mechanical enucleation procedure is inefficient and different techniques have to be used depending on the adherence properties of the cell. To circumvent these difficulties, we developed a chemical enucleation method that can have wide applicability for the production of transmitochondrial cybrids. The method is based on the use of actinomycin D to render the nuclear genome transcription/replication inactive and unable to recover after treatment. Such treated cells are fused to cells devoid of mitochondrial DNA and selected for the presence of a functional oxidative phosphorylation system. Our results showed that 95% of the clones recovered by this procedure are true transmitochondrial cybrids. This method greatly facilitates the production of transmitochondrial cybrids, thereby increasing the number of mtDNA mutations and the recipient cell types that can be studied by this system. PMID- 12907749 TI - Ligation of high-melting-temperature 'clamp' sequence extends the scanning range of rare point-mutational analysis by constant denaturant capillary electrophoresis (CDCE) to most of the human genome. AB - Mutations cause or influence the prevalence of many diseases. In human tissues, somatic point mutations have been observed at fractions at or below 4/10,000 and 5/100,000 in mitochondrial and nuclear DNA, respectively. In human populations, fractions for the multiple alleles that code for recessive deleterious syndromes are not expected to exceed 5 x 10(-4). Both nuclear and mitochondrial point mutations have been measured in human cells and tissues at fractions approaching 10(-6) using constant denaturant capillary electrophoresis (CDCE) coupled with high-fidelity PCR (hifiPCR). However, this approach is only applicable to those target sequences (approximately 100 bp) juxtaposed with a 'clamp', a higher melting-temperature sequence, in genomic DNA; such naturally clamped targets represent approximately 9% of the human genome. To open up most of the human genome to rare point-mutational analysis, a high-efficiency DNA ligation procedure was recently developed so that a clamp could be attached to any target of interest. We coupled this ligation procedure with prior CDCE/hifiPCR and achieved a sensitivity of 2 x 10(-5) in human cells for the first time using an externally attached clamp. At this sensitivity, somatic mutations, each representing an anatomically distinct cluster of cells (turnover unit) derived from a mutant stem cell, may be detected in a series of tissue samples, each containing as many as 5 x 10(4) turnover units. Additionally, rare inherited mutations may be scanned in pooled DNA samples, each derived from as many as 10(5) persons. PMID- 12907751 TI - Regionalized GC content of template DNA as a predictor of PCR success. AB - A set of 1438 human exons was subjected to nested PCR. The initial success rate using a standard PCR protocol required for ligation-independent cloning was 83.4%. Logistic regression analysis was conducted on 27 primer- and template related characteristics, of which most could be ignored apart from those related to the GC content of the template. Overall GC content of the template was a good predictor for PCR success; however, specificity and sensitivity values for predicted outcome were improved to 84.3 and 94.8%, respectively, when regionalized GC content was employed. This represented a significant improvement in predictability with respect to GC content alone (P < 0.001; chi(2)) and is expected to increase in relative sensitivity as template size increases. Regionalized GC was calculated with respect to a threshold of 61% GC content and a sliding window of 21 bp across the target sequence. Fine-tuning of PCR conditions is not practicable for all target sequences whenever a large number of genes of different lengths and GC content are to be amplified in parallel, particularly if total open reading frame or domain coverage is essential for recombinant protein synthesis. Thus, the present method is proposed as a means of grouping subsets of genes possessing potentially difficult target sequences so that PCR conditions can be optimized separately in order to obtain improved outcomes. PMID- 12907752 TI - Diminished milk synthesis in upstream stimulatory factor 2 null mice is associated with decreased circulating oxytocin and decreased mammary gland expression of eukaryotic initiation factors 4E and 4G. AB - Previous studies have suggested that upstream stimulatory factors (USFs) regulate genes involved with cell cycle progression. Because of the relationship of USFs to an important oncogene in breast cancer, c-myc, we chose to determine the importance of USF to normal mammary gland development in the mouse. Expression of USF in the mammary gland throughout development demonstrated only modest changes. Mutation of the Usf2 gene was associated with reduced fertility in females, but had no effect on prepartum mammary gland development. However, lactation performance in Usf2-/- females was only half of that observed in Usf2+/+ females, and both lactose and nitrogen were decreased in milk from Usf2-/- dams. This decrease was associated with diminished mammary tissue wet weight and luminal area by d 9 of lactation and with a decreased protein-DNA ratio. This decrease was associated with reduced abundance of the eukaryotic initiation factors eIF4E and eIF4G. Blood oxytocin concentrations on d 9 postpartum were also lower in Usf2-/- mice than Usf2+/+ mice. In contrast, the mutation had no effect on blood prolactin concentrations, mammary cell proliferation or apoptosis, mammary tissue oxytocin receptors, or milk protein gene expression. The mutation had only modest effects on maternal behavior. These data support the idea that USF is important to physiological processes necessary for the establishment and maintenance of normal lactation and suggest that USF-2 may impact lactation through both systemic and mammary cell-specific mechanisms. PMID- 12907753 TI - CCAAT/enhancer-binding protein-homologous protein expression and transcriptional activity are regulated by 3',5'-cyclic adenosine monophosphate in thyroid cells. AB - The cAMP pathway activates p38-MAPKs in the FRTL-5 rat thyroid cell line, contributing to the increased expression of the Na+/I- symporter (NIS) mRNA. This study investigates the cAMP-dependent expression and transcriptional activity of the p38-MAPK substrate CCAAT/enhancer-binding protein-homologous protein (CHOP). CHOP is expressed in the rat thyroid gland and in confluent PCCL3 and FRTL-5 cells. In FRTL-5 cells, TSH withdrawal induced a rapid down-regulation of CHOP that could be prevented by forskolin (Fk). Moreover, TSH and Fk were able to reinduce CHOP expression. The use of pharmacological inhibitors indicated that cAMP-induced CHOP expression was dependent on protein kinase A (PKA), mammalian target of rapamycin pathway, and reactive oxygen species. Transfection of a CHOP trans- reporting system revealed strong stimulation of the transcriptional activity of CHOP by Fk, by chlorophenylthio-cAMP, and by the catalytic subunit of PKA. CHOP transcriptional activity was significantly reduced by the p38-MAPK inhibitor SB203580, by transfection of a dominant-negative variant of p38alpha MAPK, or by mutation of two serine residues in CHOP targeted by p38-MAPKs. Finally, cAMP-induced NIS mRNA expression was higher in FRTL-5 cells stably transfected with CHOP cDNA than in control cells. Likewise, the activity of the NIS promoter was higher in cells overexpressing CHOP than in control cells. These findings suggest that the stimulation of CHOP expression and transcriptional activity by the cAMP pathway may contribute to the regulation of genes involved in thyroid cell differentiation. PMID- 12907754 TI - Src mediates prolactin-dependent proliferation of T47D and MCF7 cells via the activation of focal adhesion kinase/Erk1/2 and phosphatidylinositol 3-kinase pathways. AB - Prolactin (PRL) stimulates breast cancer cell proliferation; however, the involvement of PRL-activated signaling molecules in cell proliferation is not fully established. Here we studied the role of c-Src on PRL-stimulated proliferation of T47D and MCF7 breast cancer cells. We initially observed that PRL-dependent activation of focal adhesion kinase (Fak), Erk1/2, and cell proliferation was mediated by c-Src in T47D cells, because expression of a dominant-negative form of c-Src (SrcDM, K295A/Y527F) blocked the PRL-dependent effects. The Src inhibitor PP1 abrogated PRL-dependent in vivo activation of Fak, Erk1/2, p70S6K, and Akt and the proliferation of T47D and MCF7 cells; Janus kinase 2 (Jak2) activation was not affected. However, in vitro, Fak and Jak2 kinases were not directly inhibited by PP1, demonstrating the effect of PP1 on c Src kinase as an upstream activator of Fak. Expression of Fak mutant Y397F abrogated PRL-dependent activation of Fak, Erk1/2, and thymidine incorporation, but had no effect on p70S6K and Akt kinases. MAPK kinase 1/2 (Mek1/2) inhibitor PD184352 blocked PRL-induced stimulation of Erk1/2 and cell proliferation; however, p70S6K and Akt activation were unaffected. The phosphatidylinositol 3 kinase (PI3K) inhibitor LY294002 abolished cell proliferation and activation of p70S6K and Akt; however, PRL-dependent activation of Erk1/2 was not modified. Moreover, we show that both c-Src/PI3K and c-Src/Fak/Erk1/2 pathways are involved in the up-regulation of c-myc and cyclin d1 expression mediated by PRL. The previous findings suggest the existence of two PRL-dependent signaling cascades, initiated by the c-Src-mediated activation of Fak/Erk1/2 and PI3K pathways that, subsequently, control the expression of c-Myc and cyclin D1 and the proliferation of T47D and MCF7 breast cancer cells. PMID- 12907755 TI - Identification of protein tyrosine phosphatases with specificity for the ligand activated growth hormone receptor. AB - Protein tyrosine phosphatases (PTPs) play key roles in switching off tyrosine phosphorylation cascades, such as initiated by cytokine receptors. We have used substrate-trapping mutants of a large set of PTPs to identify members of the PTP family that have substrate specificity for the phosphorylated human GH receptor (GHR) intracellular domain. Among 31 PTPs tested, T cell (TC)-PTP, PTP-beta, PTP1B, stomach cancer-associated PTP 1 (SAP-1), Pyst-2, Meg-2, and PTP-H1 showed specificity for phosphorylated GHR that had been produced by coexpression with a kinase in bacteria. We then used GH-induced, phosphorylated GH receptor, purified from overexpressing mammalian cells, in a Far Western-based approach to test whether these seven PTPs were also capable of recognizing ligand-induced, physiologically phosphorylated GHR. In this assay, only TC-PTP, PTP1B, PTP-H1, and SAP-1 interacted with the mature form of the phosphorylated GHR. In parallel, we show that these PTPs recognize very different subsets of the seven GHR tyrosines that are potentially phosphorylated. Finally, mRNA tissue distribution of these PTPs by RT-PCR analysis and coexpression of the wild-type PTPs to test their ability to dephosphorylate ligand-activated GHR suggest PTP-H1 and PTP1B as potential candidates involved in GHR signaling. PMID- 12907756 TI - Compromised reproductive function in adult female mice selectively expressing mutant ErbB-1 tyrosine kinase receptors in astroglia. AB - The ErbB-1 tyrosine kinase receptor plays critical roles in regulating physiological functions. This receptor-mediated signaling in astroglia has been implicated in controlling female sexual development via activating neurons that release LH-releasing hormone (LHRH), the neuropeptide required for the secretion of LH. It remains unknown whether astroglial ErbB-1 receptors are necessary for maintaining normal adult reproductive function. Here we provide genetic evidence that astroglia-specific and time-controlled disruption of ErbB-1 receptor signaling by expressing mutant ErbB-1 receptors leads to compromised reproduction due to alteration in LHRH neuron-controlled secretion of LH in adult female mice. Therefore, astroglial ErbB-1 receptors are required for controlling LHRH neuronal function and thus maintaining adult reproduction, suggesting that compromised astroglial ErbB-1 signaling may also contribute to reproductive abnormalities in aging females. PMID- 12907757 TI - High constitutive signaling of the ghrelin receptor--identification of a potent inverse agonist. AB - Ghrelin is a GH-releasing peptide that also has an important role as an orexigenic hormone-stimulating food intake. By measuring inositol phosphate turnover or by using a reporter assay for transcriptional activity controlled by cAMP-responsive elements, the ghrelin receptor showed strong, ligand-independent signaling in transfected COS-7 or human embryonic kidney 293 cells. Ghrelin and a number of the known nonpeptide GH secretagogues acted as agonists stimulating inositol phosphate turnover further. In contrast, the low potency ghrelin antagonist, [D-Arg1,D-Phe5,D-Trp7,9,Leu11]-substance P was surprisingly found to be a high potency (EC50 = 5.2 nm) full inverse agonist as it decreased the constitutive signaling of the ghrelin receptor down to that observed in untransfected cells. The homologous motilin receptor functioned as a negative control as it did not display any sign of constitutive activity; however, upon agonist stimulation the motilin receptor signaled as strongly as the unstimulated ghrelin receptor. It is concluded that the ghrelin receptor is highly constitutively active and that this activity could be of physiological importance in its role as a regulator of both GH secretion and appetite control. It is suggested that inverse agonists for the ghrelin receptor could be particularly interesting for the treatment of obesity. PMID- 12907758 TI - Postendocytotic trafficking of the follicle-stimulating hormone (FSH)-FSH receptor complex. AB - Although the fates of the internalized hormone-receptor complexes formed by the lutropin/choriogonadotropin and the TSH receptors have been examined in some detail, much less is known about the fate of the internalized FSH-FSH receptor (FSHR) complex. Using biochemical and imaging approaches we show here that the majority of the internalized FSH-FSHR complex accumulates in endosomes and subsequently recycles back to the cell surface where the bound, intact hormone dissociates back into the medium. Only small amounts of FSH and the FSHR are routed to a lysosomal degradation pathway, and the extent of FSH-induced down regulation of the cell surface and total FSHR is minimal. This pathway was detected in heterologous (human kidney 293T) cells transfected with the rat (r) or human (h) FSHR as well as in a mouse Sertoli cell line (MSC-1) or a mouse granulosa cell line (KK-1) transfected with the rFSHR.Additional experiments using a series of C-terminal deletions of the rFSHR and the hFSHR showed that the recycling of the internalized FSH-FSHR complex and the extent of hFSH-induced down-regulation is dictated by a short stretch of amino acids present at the extreme C-terminal end of the receptor.We conclude that most of the internalized FSH-FSHR complex is recycled back to the cell surface, that this recycling pathway is highly dependent on amino acid residues present near the C terminus of the FSHR, and that it is an important determinant of the extent of down regulation of the FSHR. PMID- 12907759 TI - Epitope map for a growth hormone receptor agonist monoclonal antibody, MAb 263. AB - Monoclonal antibody (MAb) 263 is a widely used monoclonal antibody that recognizes the extracellular domain (ECD) of the GH receptor. It has been shown to act as a GH agonist both in vitro and in vivo, and we report here that it must be divalent to exert its effect on the full-length receptor. To understand the mechanism of its agonist action, we have determined the precise epitope for this antibody using a novel random PCR mutagenesis approach together with expression screening in yeast. A library of 5200 clones of rabbit GH receptor ECD mutants were screened both with MAb 263 and with an anticarboxy-tag antibody to verify complete ECD expression. Sequencing for clones that expressed complete ECD but were not MAb 263 positive identified 20 epitope residues distributed in a discontinuous manner throughout the ECD. The major part of the epitope, as revealed after mapping onto the crystal structure model of the ECD molecule, was located on the side and upper portion of domain 1, particularly within the D-E strand disulfide loop 79-96. Molecular dynamics docking of an antibody of the same isotype as MAb 263 was used to dock the bivalent antibody to the 1528-A2 epitope and to visualize the likely consequences of MAb binding. The minimized model enables the antibody to grasp two receptors in a pincer-like movement from opposite sides, facilitating alignment of the receptor dimerization domains in a manner similar to, but not identical with, GH. PMID- 12907760 TI - Hypothyroidism in thyroid transcription factor 1 haploinsufficiency is caused by reduced expression of the thyroid-stimulating hormone receptor. AB - Humans expressing one allele of the thyroid transcription factor 1 (TTF1) gene have neurological symptoms and increased serum TSH with variable degrees of hypothyroidism. Ttf1+/- mice have also poor coordination and increased serum TSH concentration (205 +/- 22 vs. 92 +/- 12 mU/liter; P < 0.001) and slightly lower T4 (46 +/- 3 vs. 63 +/- 6 nmol/liter; P < 0.02) as compared with Ttf1+/+ mice. To determine whether the hypothyroidism is of central or primary origin, we examined the bioactivity of TSH, thyroidal response to exogenous TSH and the expression of genes regulated by TTF1. TSH bioactivity was normal, but T4 response to a low but not high dose of TSH was significantly reduced in the Ttf1+/- mice (5.5 +/- 2.2 vs. 15.3 +/- 4.1 nmol/liter; P < 0.03), indicating a reduced thyroidal response. Thyroid mRNAs were measured by real-time PCR (Ttf1+/+ littermates = 100%). Ttf1+/ mice had half the levels of TTF1 mRNA (54 +/- 9; P < 0.01) and protein, confirming their haploinsufficiency. Significantly lower levels of mRNAs were observed for two of the three genes with TTF1 cis elements: TSH receptor (TSHr, 57 +/- 4%; P < 0.002), thyroglobulin (63 +/- 7%; P < 0.005), but not thyroid peroxidase (81 +/- 12%; P > 0.05). No significant difference between the two genotypes was found for Pax8, sodium iodide symporter, and iodothyronine deiodinase 1. These results show that Ttf1 haploinsufficiency causes a reduction in the expression of TSHr and thyroglobulin, genes with TTF1 binding sites in their promoter regions. The low TSHr is only partially compensated by an increase in TSH secretion because T4 remains mildly reduced. However, administration of a larger amount of TSH obliterates the response differences by saturating a reduced amount of receptor. PMID- 12907762 TI - Multiple authors, multiple problems. PMID- 12907761 TI - TCF and Groucho-related genes influence pituitary growth and development. AB - Mutations in the prophet of PIT1 gene (PROP1) are the most common cause of multiple pituitary hormone deficiency in humans; however, the mechanism of PROP1 action is not well understood. We report that Prop1 is essential for dorsally restricted expression of a Groucho-related gene, transducin-like enhancer of split 3 (Tle3), which encodes a transcriptional corepressor. Deficiency of a related gene, amino terminal enhancer of split (Aes), causes pituitary anomalies and growth insufficiency. TLE3 and AES have been shown to interact with TCF/LEF (transcripiton factors of the T cell-specific and lymphoid enhancer specific group) family members in cell culture systems. In the absence of TCF4 (Tcf7L2), Prop1 levels are elevated, pituitary hyperplasia ensues and palate closure is abnormal. Thus, we demonstrate that Tcf4 and Aes influence pituitary growth and development, and place Tcf4 and Tle3 in the genetic hierarchy with Prop1. PMID- 12907763 TI - Radiation science. Sparks fly over new report on Hiroshima bomb radiation. PMID- 12907764 TI - France. Basic research chief given the boot. PMID- 12907765 TI - Planetary science. Sniffing out Martian hospitality. PMID- 12907767 TI - Paleontology. Ancient trackways in strip mine threatened by reburial. PMID- 12907766 TI - Evolution. Passages found through labyrinth of bacterial evolution. PMID- 12907768 TI - Relativity theory. Souped-up Archimedes equation torpedoes submarine paradox. PMID- 12907769 TI - National security. 'Terrorism futures' could have a future, experts say. PMID- 12907770 TI - Atomic physics. Ultracold atoms spark a hot race. PMID- 12907771 TI - Atomic physics. The contenders. PMID- 12907772 TI - Scientific workforce. Poor job market blunts impact of new master's programs. PMID- 12907773 TI - Robotics. Shape shifters tread a daunting path toward reality. PMID- 12907774 TI - Robotics. Topologists and roboticists explore an 'inchoate world'. PMID- 12907775 TI - Neuroscience. Depression drugs' powers may rest on new neurons. PMID- 12907776 TI - Jean-Jacques Dordain profile: applying himself to the business of space. PMID- 12907777 TI - A shortage of Ph.D.s? PMID- 12907778 TI - Another perspective on hominid diversity. PMID- 12907779 TI - Chromosomal speciation. PMID- 12907780 TI - Planetary science. Human space exploration is about more than just science. PMID- 12907781 TI - Oceans. Sounding out ocean fine structure. PMID- 12907782 TI - Social science. Ignorance, knowledge, and outcomes in a small world. PMID- 12907784 TI - Climate. Taking the pulse of the tropical water cycle. PMID- 12907783 TI - Development. Longing for ligand: hedgehog, patched, and cell death. PMID- 12907786 TI - Bacterial chromosome dynamics. AB - Bacterial chromosomes are highly compacted structures and share many properties with their eukaryote counterparts, despite not being organized into chromatin or being contained within a cell nucleus. Proteins conserved across all branches of life act in chromosome organization, and common mechanisms maintain genome integrity and ensure faithful replication. The principles that underlie chromosome segregation in bacteria and eukaryotes share similarities, although bacteria segregate DNA as it replicates and lack a eukaryote-like mitotic apparatus for segregating chromosomes. This may be because the distances that newly replicated bacterial chromosomes move apart before cell division are small as compared to those in eukaryotes. Bacteria specify positional information, which determines where cell division will occur and which places the replication machinery and chromosomal loci at defined locations that change during cell cycle progression. PMID- 12907787 TI - Chromosome choreography: the meiotic ballet. AB - The separation of homologous chromosomes during meiosis in eukaryotes is the physical basis of Mendelian inheritance. The core of the meiotic process is a specialized nuclear division (meiosis I) in which homologs pair with each other, recombine, and then segregate from each other. The processes of chromosome alignment and pairing allow for homolog recognition. Reciprocal meiotic recombination ensures meiotic chromosome segregation by converting sister chromatid cohesion into mechanisms that hold homologous chromosomes together. Finally, the ability of sister kinetochores to orient to a single pole at metaphase I allows the separation of homologs to two different daughter cells. Failures to properly accomplish this elegant chromosome dance result in aneuploidy, a major cause of miscarriage and birth defects in human beings. PMID- 12907788 TI - Prokaryotic chromosomes and disease. AB - Recent insights into bacterial genome organization and function have improved our understanding of the nature of pathogenic bacteria and their ability to cause disease. It is becoming increasingly clear that the bacterial chromosome constantly undergoes structural changes due to gene acquisition and loss, recombination, and mutational events that have an impact on the pathogenic potential of the bacterium. Even though the bacterial genome includes additional genetic elements, the chromosome represents the most important entity in this context. Here, we will show that various processes of genomic instability have an influence on the many manifestations of infectious disease. PMID- 12907789 TI - Structural dynamics of eukaryotic chromosome evolution. AB - Large-scale genome sequencing is providing a comprehensive view of the complex evolutionary forces that have shaped the structure of eukaryotic chromosomes. Comparative sequence analyses reveal patterns of apparently random rearrangement interspersed with regions of extraordinarily rapid, localized genome evolution. Numerous subtle rearrangements near centromeres, telomeres, duplications, and interspersed repeats suggest hotspots for eukaryotic chromosome evolution. This localized chromosomal instability may play a role in rapidly evolving lineage specific gene families and in fostering large-scale changes in gene order. Computational algorithms that take into account these dynamic forces along with traditional models of chromosomal rearrangement show promise for reconstructing the natural history of eukaryotic chromosomes. PMID- 12907790 TI - Heterochromatin and epigenetic control of gene expression. AB - Eukaryotic DNA is organized into structurally distinct domains that regulate gene expression and chromosome behavior. Epigenetically heritable domains of heterochromatin control the structure and expression of large chromosome domains and are required for proper chromosome segregation. Recent studies have identified many of the enzymes and structural proteins that work together to assemble heterochromatin. The assembly process appears to occur in a stepwise manner involving sequential rounds of histone modification by silencing complexes that spread along the chromatin fiber by self-oligomerization, as well as by association with specifically modified histone amino-terminal tails. Finally, an unexpected role for noncoding RNAs and RNA interference in the formation of epigenetic chromatin domains has been uncovered. PMID- 12907791 TI - Bacterial sex: playing voyeurs 50 years later. AB - The concept of chromosomes with a ring structure was born during the early studies of bacterial sexuality, and the discovery of fertility factors- episomes or plasmids-provided much later the key tools for gene cloning and biotechnology. But the plasmid-mediated transfer of antibiotic and other resistances, as well as pathogenicity, has served bacteria well in their own adaptive evolution. PMID- 12907792 TI - Measles outbreaks in a population with declining vaccine uptake. PMID- 12907793 TI - Requirement of hippocampal neurogenesis for the behavioral effects of antidepressants. AB - Various chronic antidepressant treatments increase adult hippocampal neurogenesis, but the functional importance of this phenomenon remains unclear. Here, using genetic and radiological methods, we show that disrupting antidepressant-induced neurogenesis blocks behavioral responses to antidepressants. Serotonin 1A receptor null mice were insensitive to the neurogenic and behavioral effects of fluoxetine, a serotonin selective reuptake inhibitor. X-irradiation of a restricted region of mouse brain containing the hippocampus prevented the neurogenic and behavioral effects of two classes of antidepressants. These findings suggest that the behavioral effects of chronic antidepressants may be mediated by the stimulation of neurogenesis in the hippocampus. PMID- 12907794 TI - An all-optical quantum gate in a semiconductor quantum dot. AB - We report coherent optical control of a biexciton (two electron-hole pairs), confined in a single quantum dot, that shows coherent oscillations similar to the excited-state Rabi flopping in an isolated atom. The pulse control of the biexciton dynamics, combined with previously demonstrated control of the single exciton Rabi rotation, serves as the physical basis for a two-bit conditional quantum logic gate. The truth table of the gate shows the features of an all optical quantum gate with interacting yet distinguishable excitons as qubits. Evaluation of the fidelity yields a value of 0.7 for the gate operation. Such experimental capability is essential to a scheme for scalable quantum computation by means of the optical control of spin qubits in dots. PMID- 12907795 TI - Compensation of the birefringence of a polymer by a birefringent crystal. AB - We report a method for compensating the birefringence of optical polymers by doping them with inorganic birefringent crystals. In this method, an inorganic birefringent material is chosen that has the opposite birefringence to that of the polymer and has rod-shaped crystals that are oriented when the polymer chains are oriented. The birefringence of the polymer is thus compensated by the opposing birefringence of the crystal. Birefringence is minimized in various polymer optical devices by adjusting process conditions, because it degrades the performance of devices. This method minimizes it, independent of process conditions, which potentially improves the productivity of devices. PMID- 12907796 TI - Catalytic, oxidative condensation of CH4 to CH3COOH in one step via CH activation. AB - Acetic acid is an important petrochemical that is currently produced from methane (or coal) in a three-step process based on carbonylation of methanol. We report a direct, selective, oxidative condensation of two methane molecules to acetic acid at 180 degrees C in liquid sulfuric acid. Carbon-13 isotopic labeling studies show that both carbons of acetic acid originate from methane. The reaction is catalyzed by palladium, and the results are consistent with the reaction occurring by tandem catalysis, involving methane C-H activation to generate Pd CH3 species, followed by efficient oxidative carbonylation with methanol, generated in situ from methane, to produce acetic acid. PMID- 12907797 TI - Synthesis of zeolite as ordered multicrystal arrays. AB - Zeolites are crystalline nanoporous aluminosilicates widely used in industry. In order for zeolites to find applications as innovative materials, they need to be organized into large two- and three-dimensional (2D and 3D) arrays. We report that uniformly aligned polyurethane films can serve as templates for the synthesis of uniformly aligned 2D and possibly 3D arrays of silicalite-1 crystals, in which the orientations of the crystals are controlled by the nature of the polymers. We propose that the supramolecularly organized organic-inorganic composites that consist of the hydrolyzed organic products and the seed crystals are responsible for this phenomenon. PMID- 12907798 TI - Thermohaline fine structure in an oceanographic front from seismic reflection profiling. AB - We present acoustic images of oceanic thermohaline structure created from marine seismic reflection profiles across the major oceanographic front between the Labrador Current and the North Atlantic Current. The images show that distinct water masses can be mapped, and their internal structure imaged, using low frequency acoustic reflections from sound speed contrasts at interfaces across which temperature changes. The warm/cold front is characterized by east-dipping reflections generated by thermohaline intrusions in the uppermost 1000 meters of the ocean. Our results imply that marine seismic reflection techniques can provide excellent spatial resolution of important oceanic phenomena, including thermohaline intrusions, internal waves, and eddies. PMID- 12907799 TI - Observation of long supershear rupture during the magnitude 8.1 Kunlunshan earthquake. AB - The 2001 Kunlunshan earthquake was an extraordinary event that produced a 400-km long surface rupture. Regional broadband recordings of this event provide an opportunity to accurately observe the speed at which a fault ruptures during an earthquake, which has important implications for seismic risk and for understanding earthquake physics. We determined that rupture propagated on the 400-km-long fault at an average speed of 3.7 to 3.9 km/s, which exceeds the shear velocity of the brittle part of the crust. Rupture started at sub-Rayleigh wave velocity and became supershear, probably approaching 5 km/s, after about 100 km of propagation. PMID- 12907800 TI - An experimental study of search in global social networks. AB - We report on a global social-search experiment in which more than 60,000 e-mail users attempted to reach one of 18 target persons in 13 countries by forwarding messages to acquaintances. We find that successful social search is conducted primarily through intermediate to weak strength ties, does not require highly connected "hubs" to succeed, and, in contrast to unsuccessful social search, disproportionately relies on professional relationships. By accounting for the attrition of message chains, we estimate that social searches can reach their targets in a median of five to seven steps, depending on the separation of source and target, although small variations in chain lengths and participation rates generate large differences in target reachability. We conclude that although global social networks are, in principle, searchable, actual success depends sensitively on individual incentives. PMID- 12907801 TI - Phylogenetics and the cohesion of bacterial genomes. AB - Gene acquisition is an ongoing process in many bacterial genomes, contributing to adaptation and ecological diversification. Lateral gene transfer is considered the primary explanation for discordance among gene phylogenies and as an obstacle to reconstructing the tree of life. We measured the extent of phylogenetic conflict and alien-gene acquisition within quartets of sequenced genomes. Although comparisons of complete gene inventories indicate appreciable gain and loss of genes, orthologs available for phylogenetic reconstruction are consistent with a single tree. PMID- 12907802 TI - Nervous system targets of RNA editing identified by comparative genomics. AB - An unknown number of precursor messenger RNAs undergo genetic recoding by modification of adenosine to inosine, a reaction catalyzed by the adenosine deaminases acting on RNA (ADARs). Discovery of these edited transcripts has always been serendipitous. Using comparative genomics, we identified a phylogenetic signature of RNA editing. We report the identification and experimental verification of 16 previously unknown ADAR target genes in the fruit fly Drosophila and one in humans-more than the sum total previously reported. All of these genes are involved in rapid electrical and chemical neurotransmission, and many of the edited sites recode conserved and functionally important amino acids. These results point to a pivotal role for RNA editing in nervous system function. PMID- 12907803 TI - Single molecule profiling of alternative pre-mRNA splicing. AB - Alternative pre-messenger RNA splicing is an important mechanism for generating protein diversity and may explain in part how mammalian complexity arises from a surprisingly small complement of genes. Here, we describe "digital polony exon profiling,"a single molecule-based technology for studying complex alternative pre-messenger RNA splicing. This technology allows researchers to monitor the combinatorial diversity of exon inclusion in individual transcripts. A minisequencing strategy provides single nucleotide resolution, and the digital nature of the technology allows quantitation of individual splicing variants. Digital polony exon profiling can be used to investigate the physiological and pathological roles of alternately spliced messenger RNAs, as well as the mechanisms by which these messenger RNAs are produced. PMID- 12907804 TI - Retrograde viral delivery of IGF-1 prolongs survival in a mouse ALS model. AB - Amyotrophic lateral sclerosis (ALS) is a progressive, lethal neuromuscular disease that is associated with the degeneration of spinal and brainstem motor neurons, leading to atrophy of limb, axial, and respiratory muscles. The cause of ALS is unknown, and there is no effective therapy. Neurotrophic factors are candidates for therapeutic evaluation in ALS. Although chronic delivery of molecules to the central nervous system has proven difficult, we recently discovered that adeno-associated virus can be retrogradely transported efficiently from muscle to motor neurons of the spinal cord. We report that insulin-like growth factor 1 prolongs life and delays disease progression, even when delivered at the time of overt disease symptoms. PMID- 12907805 TI - Inhibition of neuroepithelial patched-induced apoptosis by sonic hedgehog. AB - During early development in vertebrates, Sonic hedgehog (Shh) is produced by the notochord and the floor plate. A ventrodorsal gradient of Shh directs ventrodorsal patterning of the neural tube. However, Shh is also required for the survival of neuroepithelial cells. We show that Patched (Ptc) induces apoptotic cell death unless its ligand Shh is present to block the signal. Moreover, the blockade of Ptc-induced cell death partly rescues the chick spinal cord defect provoked by Shh deprivation. Thus, the proapoptotic activity of unbound Ptc and the positive effect of Shh-bound Ptc on cell differentiation probably cooperate to achieve the appropriate spinal cord development. PMID- 12907806 TI - Amygdalar and hippocampal theta rhythm synchronization during fear memory retrieval. AB - The amygdalohippocampal circuit plays a pivotal role in Pavlovian fear memory. We simultaneously recorded electrical activity in the lateral amygdala (LA) and the CA1 area of the hippocampus in freely behaving fear-conditioned mice. Patterns of activity were related to fear behavior evoked by conditioned and indifferent sensory stimuli and contexts. Rhythmically synchronized activity at theta frequencies increased between the LA and the CA1 after fear conditioning and became significant during confrontation with conditioned fear stimuli and expression of freezing behavior. Synchronization of theta activities in the amygdalohippocampal network represents a neuronal correlate of conditioned fear, apt to improve neuronal communication during memory retrieval. PMID- 12907808 TI - Dissipationless quantum spin current at room temperature. AB - Although microscopic laws of physics are invariant under the reversal of the arrow of time, the transport of energy and information in most devices is an irreversible process. It is this irreversibility that leads to intrinsic dissipations in electronic devices and limits the possibility of quantum computation. We theoretically predict that the electric field can induce a substantial amount of dissipationless quantum spin current at room temperature, in hole-doped semiconductors such as Si, Ge, and GaAs. On the basis of a generalization of the quantum Hall effect, the predicted effect leads to efficient spin injection without the need for metallic ferromagnets. Principles found here could enable quantum spintronic devices with integrated information processing and storage units, operating with low power consumption and performing reversible quantum computation. PMID- 12907807 TI - Major ecological transitions in wild sunflowers facilitated by hybridization. AB - Hybridization is frequent in many organismal groups, but its role in adaptation is poorly understood. In sunflowers, species found in the most extreme habitats are ancient hybrids, and new gene combinations generated by hybridization are speculated to have contributed to ecological divergence. This possibility was tested through phenotypic and genomic comparisons of ancient and synthetic hybrids. Most trait differences in ancient hybrids could be recreated by complementary gene action in synthetic hybrids and were favored by selection. The same combinations of parental chromosomal segments required to generate extreme phenotypes in synthetic hybrids also occurred in ancient hybrids. Thus, hybridization facilitated ecological divergence in sunflowers. PMID- 12907809 TI - Galaxy disruption in a halo of dark matter. AB - The relics of disrupted satellite galaxies have been found around the Milky Way and Andromeda, but direct evidence of a satellite galaxy in the early stages of disruption has remained elusive. We have discovered a dwarf satellite galaxy in the process of being torn apart by gravitational tidal forces as it merges with a larger galaxy's dark matter halo. Our results illustrate the morphological transformation of dwarf galaxies by tidal interaction and the continued buildup of galaxy halos. PMID- 12907810 TI - Coronary artery imaging with multidetector CT: visualization issues. AB - Noninvasive imaging of the coronary arteries has attracted growing interest in the past few years. One of the possible acquisition techniques is multidetector computed tomography (CT) that produces large three-dimensional (3D) data sets that require visualization techniques for data evaluation. The objective of this article is to increase knowledge of possible 3D visualization techniques together with their advantages and disadvantages for the routine evaluation of cardiac data sets. Common imaging techniques available to the radiologist at standard workstations are multiplanar reformation (MPR), oblique MPR, curved MPR, maximum intensity projection (MIP), shaded-surface display, and direct volume rendering. Each of these techniques has its advantages and disadvantages for the visualization of the coronary artery tree. Several additions to the basic techniques have been developed to overcome some of their shortcomings. Different clinical examinations, such as stent evaluation, stenosis evaluation, and bypass evaluation, require different visualization techniques. The choice of preferred technique for each clinical study depends on the advantages and disadvantages of the various techniques as described in the literature. Because of the large number of possible settings and projection angles, it is important for users to interactively manipulate the images and review the whole vessel volume rather than just looking at static reformatted images. Errors such as findings of false stenoses can be avoided by means of accurate and appropriate use of software features. This requires training of users both with regard to the capabilities of the software and the background of the different techniques and their possible pitfalls. The authors believe that volume rendering of the whole heart is useful for anatomic evaluation of the coronary arteries. For more detailed observation of specific lesions, slab imaging with volume rendering or MIP is required. PMID- 12907811 TI - Fiber-tracking method reveals sensorimotor pathway involvement in stroke patients. AB - BACKGROUND AND PURPOSE: We tested the feasibility of a new MRI technique that provides visualization of the sensorimotor tracts in vivo in a group of stroke victims. SUMMARY OF REPORT: Fourteen patients with small infarctions involving the white matter of the supratentorial brain were evaluated. Sensorimotor tracts on the lesional and contralesional sides were successfully depicted in all cases. The position of the sensorimotor tracts relative to the infarct was in good agreement with clinical symptoms. The overall sensitivity and specificity for sensorimotor tract involvement were 100% and 77%, respectively. CONCLUSIONS: Our proposed fiber-tracking method was shown to be a clinically feasible technique that correlates well with clinical symptoms. PMID- 12907812 TI - Antibiotics in primary prevention of stroke in the elderly. AB - BACKGROUND AND PURPOSE: An increasing number of reports have linked infections to atherosclerosis and thrombosis. Thus, use of antibiotics may lower the risk of developing cerebrovascular disease. We investigated whether antibiotic use is associated with the risk of stroke in elderly individuals treated for hypertension. METHODS: A cohort of 29 937 elderly subjects initiating antihypertensive therapy between 1982 and 1995 was formed from the Quebec healthcare insurance database. A nested case-control design was used in which each subject hospitalized with a primary discharge diagnosis of stroke between 1987 and 1995 was matched on calendar time to 5 randomly selected controls from the cohort. Conditional logistic regression was used to estimate odds ratios of stroke after adjustment for predisposing factors. RESULTS: We identified 1888 cases and 9440 controls. The overall adjusted odds ratio for current antibiotic use was 0.80 (95% confidence interval, 0.63 to 1.01), and that for recent use was 0.81 (95% confidence interval, 0.70 to 0.94). Penicillin was the only individual antibiotic class that showed a protective association across different time windows. No significant association was found between stroke risk and the use of fluoroquinolones, macrolides, tetracyclines, or cephalosporins. CONCLUSIONS: Although no clear, consistent associations between overall antibiotic use and cerebrovascular disease could be found, an intriguing association between penicillin use and stroke should be explored further. PMID- 12907813 TI - Endothelial and platelet activation in acute ischemic stroke and its etiological subtypes. AB - BACKGROUND AND PURPOSE: Activation of endothelial cells and platelets is an important mediator of atherothrombosis. Markers of endothelial cell and platelet activation such as soluble adhesion molecules can be measured in plasma. We hypothesized that patients with acute ischemic stroke would have increased blood concentrations of soluble E-selectin and von Willebrand factor (vWF), primarily reflecting activation of endothelial cells, and increased concentrations of soluble P-selectin and platelet-derived microvesicles (PDM), primarily reflecting activation of platelets, compared with healthy controls. We also hypothesized that these markers would be differentially elevated in ischemic stroke caused by large- and small-artery atherothrombosis compared with cardiogenic embolism. METHODS: We conducted a case-control study of 200 hospital-referred cases of first-ever ischemic stroke and 205 randomly selected community controls stratified by age, sex, and postal code. Using established criteria, we classified cases of stroke by etiological subtype in a blinded fashion. The prevalence of vascular risk factors and blood concentrations of E-selectin, P selectin, vWF antigen, and PDM were determined in stroke cases within 7 days and at 3 to 6 months after stroke and in controls. RESULTS: Mean blood concentrations of soluble E-selectin, P-selectin, and PDM within 7 days of stroke onset were all significantly higher in cases compared with controls. At 3 to 6 months after stroke, the mean blood concentrations of E-selectin and P-selectin fell significantly below that of controls, and PDM concentrations remained elevated. There was a strong, graded, and independent (of age, sex, and vascular risk factors) association between increasing blood concentrations of E-selectin during the acute phase and all etiological subtypes of ischemic stroke, particularly ischemic stroke caused by large-artery atherothrombosis. There was also a significant, graded, and independent association between increasing blood concentrations of vWF during the acute phase and ischemic stroke caused by large artery atherothrombosis. CONCLUSIONS: We have demonstrated significant associations between acute elevation of blood markers of endothelial cell and platelet activation and ischemic stroke and between acute elevation of blood markers of endothelial cell activation and ischemic stroke caused by large-artery atherothrombosis. Persistent elevated blood concentrations of PDM may be a marker of increased risk of ischemic stroke. PMID- 12907814 TI - Impact of hospital-related factors on outcome after treatment of cerebral aneurysms. AB - BACKGROUND AND PURPOSE: The goal of this study was to examine the impact of hospital characteristics on outcome after the treatment of ruptured and unruptured cerebral aneurysms. METHODS: We identified all discharges in New York State from 1995 through 2000 with a principal diagnosis of subarachnoid hemorrhage (SAH) or unruptured cerebral aneurysm (UCA) in patients who were treated by aneurysm clipping, wrapping, or endovascular coiling. An adverse outcome was defined as in-hospital death or discharge to a rehabilitation hospital or long-term facility. We examined the effect of hospital factors, including the rate of endovascular therapy and overall procedural volume, on outcome, length of stay, and total charges. RESULTS: There were 2200 (36.9%) and 3763 (63.1%) admissions for attempted treatment of UCA and SAH, respectively. The 10 highest-volume hospitals performed half of all the procedures. Overall, hospital volume was associated with fewer adverse outcomes and lower in-hospital mortality for both UCA (adverse outcome: odds ratio [OR], 0.89; P<0.0001; mortality: OR, 0.94; P=0.002 for each 10 additional procedures performed per year) and SAH (adverse outcome: OR, 0.94; P=0.03; mortality: OR, 0.95; P=0.005). Use of endovascular therapy (each additional 10% of cases performed endovascularly) was associated with fewer adverse outcomes after treatment of unruptured aneurysm (0.83, P=0.026). Hospital volume had more of an effect on outcome after aneurysm clipping than after endovascular therapy. CONCLUSIONS: Hospital procedural volume and the propensity of a hospital to use endovascular therapy are both independently associated with better outcome. Improvement in outcome could be achieved by a program of regionalization and selective referral for the treatment of cerebral aneurysms. PMID- 12907815 TI - Elevated plasma homocysteine was associated with hemorrhagic and ischemic stroke, but methylenetetrahydrofolate reductase gene C677T polymorphism was a risk factor for thrombotic stroke: a Multicenter Case-Control Study in China. AB - BACKGROUND AND PURPOSE: It is still controversial whether elevated plasma homocysteine and the C677T polymorphism of methylenetetrahydrofolate reductase (MTHFR) gene are risk factors for stroke. The aim of the present study was to investigate the association between the 2 factors and stroke in Chinese in a large case-control study. METHODS: We recruited 1823 stroke patients (807 cerebral thrombosis, 513 lacunar infarction, 503 intracerebral hemorrhage) and 1832 controls. Total plasma homocysteine was determined by high-performance liquid chromatography. C677T polymorphism was genotyped by polymerase chain reaction and HinfI digestion. RESULTS: Total plasma homocysteine levels were significantly higher in cases than controls (median, 14.7 versus 12.8 micromol/L; P<0.001) and associated with an increased risk of 1.87-fold (95% confidence interval [CI], 1.58 to 2.22) for overall stroke, 1.72-fold (95% CI, 1.39 to 2.12) for cerebral thrombosis, 1.89-fold (95% CI, 1.50 to 2.40) for lacunar infarction, and 1.94-fold (95% CI, 1.48 to 2.55) for intracerebral hemorrhage. The C677T mutation of the MTHFR gene was positively correlated with plasma homocysteine levels in both controls (beta=0.250, P<0.001) and cases (beta=0.272, P<0.001) and more frequently in cases than in controls (47.0% versus 44.2%, P=0.017). The TT genotype was associated with an increased risk for overall stroke (odds ratio, 1.27; 95% CI, 1.04 to 1.56) and thrombotic stroke (odds ratio, 1.37; 95% CI, 1.06 to 1.78). CONCLUSIONS: The C677T polymorphism of the MTHFR gene was associated with increased risk of cerebral thrombotic stroke in Chinese. Total plasma homocysteine was correlated with both ischemic and hemorrhagic stroke, suggesting potential initiation of homocysteine-lowering therapy in this population. PMID- 12907816 TI - Histopathological evaluation of middle cerebral artery after percutaneous intracranial transluminal angioplasty. AB - BACKGROUND: Intracranial atherosclerosis accounts for 8% to 10% of all ischemic strokes, and intracranial angioplasty is increasingly performed to treat stenotic lesions. We report an autopsy case and discuss the effects of intracranial angioplasty for atherosclerotic arteries. CASE DESCRIPTION: A 77-year-old patient died 9 days after angioplasty of the left middle cerebral artery as a result of cardiorespiratory failure. The patient was anticoagulated before, during, and after the procedure with heparin, aspirin, and clopidogrel. At the site of angioplasty, the densely fibrotic eccentric plaque was displaced from the adjacent media into the lumen, distorting it and forming elongated projections. No local thrombosis, plaque compression, or inflammation was observed. Additionally, an intramural hemorrhage extended from the site of angioplasty into the stenotic proximal inferior division of the left middle cerebral artery. CONCLUSIONS: Histopathological findings after intracranial angioplasty parallel those in other arterial territories. The implications of these pathological findings on the medical and endovascular treatment of intracranial atherosclerosis are discussed. PMID- 12907817 TI - Proportion of different subtypes of stroke in China. AB - BACKGROUND AND PURPOSE: The goal of this article is to clarify the proportion of stroke subtypes in China, where stoke is the most common cause of death. METHODS: A total of 16,031 first-ever strokes in subjects >or=25 years of age were identified in 1991 to 2000 from 17 Chinese populations through a community-based cardiovascular disease surveillance program in the China Multicenter Collaborative Study of Cardiovascular Epidemiology. World Health Organization diagnosis criteria were used for classification of stroke subtypes. RESULTS: CT scan rate of stroke cases reached a satisfactorily high level only after 1996 in the study populations. In 8268 first-ever stroke events from 10 populations with CT scan rate >75% in 1996 to 2000, 1.8% were subarachnoid hemorrhage, 27.5% were intracerebral hemorrhage, 62.4% were cerebral infarction, and 8.3% were undetermined stroke. The proportion of intracerebral hemorrhage varied from 17.1% to 39.4% and that for cerebral infarction varied from 45.5% to 75.9% from population to population. The ratio of ischemic to hemorrhagic stroke ranged from 1.1 to 3.9 and averaged 2.0). The 28-day fatality rate was 33.3% for subarachnoid hemorrhage, 49.4% for intracerebral hemorrhage, 16.9% for cerebral infarction, and 64.6% for undetermined stroke. CONCLUSIONS: In our study, ischemic stroke was more frequent and its proportion was higher than hemorrhagic stroke in Chinese populations. Although hemorrhagic stroke was more frequent in Chinese than in Western populations, the variation in the proportion of stroke subtypes among Chinese populations could be as large as or larger than that between Chinese and Western populations. PMID- 12907818 TI - Probability of regaining dexterity in the flaccid upper limb: impact of severity of paresis and time since onset in acute stroke. AB - BACKGROUND AND PURPOSE: To improve the accuracy of early postonset prediction of motor recovery in the flaccid hemiplegic arm, the effects of change in motor function over time on the accuracy of prediction were evaluated, and a prediction model for the probability of regaining dexterity at 6 months was developed. METHODS: In 102 stroke patients, dexterity and paresis were measured with the Action Research Arm Test, Motricity Index, and Fugl-Meyer motor evaluation. For model development, 23 candidate determinants were selected. Logistic regression analysis was used for prognostic factors and model development. RESULTS: At 6 months, some dexterity in the paretic arm was found in 38%, and complete functional recovery was seen in 11.6% of the patients. Total anterior circulation infarcts, right hemisphere strokes, homonymous hemianopia, visual gaze deficit, visual inattention, and paresis were statistically significant related to a poor arm function. Motricity Index leg scores of at least 25 points in the first week and Fugl-Meyer arm scores of 11 points in the second week increasing to 19 points in the fourth week raised the probability of developing some dexterity (Action Research Arm Test >or=10 points) from 74% (positive predictive value [PPV], 0.74; 95% confidence interval [CI], 0.63 to 0.86) to 94% (PPV, 0.83; 95% CI, 0.76 to 0.91) at 6 months. No change in probabilities of prediction dexterity was found after 4 weeks. CONCLUSIONS: Based on the Fugl-Meyer scores of the flaccid arm, optimal prediction of arm function outcome at 6 months can be made within 4 weeks after onset. Lack of voluntary motor control of the leg in the first week with no emergence of arm synergies at 4 weeks is associated with poor outcome at 6 months. PMID- 12907819 TI - Prospective evaluation of carotid bruit as a predictor of first stroke in type 2 diabetes: the Fremantle Diabetes Study. AB - BACKGROUND AND PURPOSE: We sought to examine prospectively the relationship between asymptomatic carotid bruit and stroke in type 2 diabetes. METHODS: We studied 1181 (91.3%) of a community-based sample of 1294 patients with type 2 diabetes. These patients had no history of cerebrovascular disease at recruitment during 1993-1996 and were followed until the end of January 2002. Hospital morbidity and death register data relating to cerebrovascular events were also available. Cox proportional hazards models were used to determine whether carotid bruit status was an independent predictor of stroke and to identify other significant cerebrovascular risk factors. RESULTS: One hundred thirty-four patients (11.3%) suffered a first stroke during 6.5+/-2.2 (mean+/-SD) years of follow-up. In the first 2 years after study entry, first stroke (n=45/1181; 3.8%) was strongly predicted by the presence of carotid bruit(s) after adjustment for known cardiovascular risk factors and other potentially confounding variables (hazard ratio, 6.7; 95% CI, 3.0 to 14.9; P<0.001). Between 2 years and census, first stroke (n=89/1083; 8.2%) was not associated with carotid bruit(s) (P=0.97). Age and diastolic blood pressure were other determinants of stroke in the first 2 years, while age, atrial fibrillation/flutter, and microalbuminuria were independent predictors of subsequent stroke. CONCLUSIONS: Type 2 diabetic patients found to have incidental carotid bruits have >6 times the risk of first stroke in the first 2 years than patients without a bruit and should receive intensified management of vascular risk factors. Carotid imaging with a view to surgical intervention in these patients remains controversial. PMID- 12907820 TI - Craniotomy for treatment of unruptured aneurysms is not associated with long-term cognitive dysfunction. AB - BACKGROUND AND PURPOSE: Many studies have reported frequent cognitive deficits associated with subarachnoid hemorrhage (SAH) and aneurysm repair. One study found more severe cognitive deficits after clipping than coiling of aneurysms, raising the possibility that deficits are due to surgery instead of SAH itself. This possibility was directly addressed by evaluating the cognitive effects of surgery without SAH. The goal of this study was to identify changes in cognitive function associated with surgical clipping of unruptured intracerebral aneurysms. METHODS: A consecutive series of 25 patients who underwent surgical clipping of >or=1 unruptured intracerebral aneurysm were tested within 1 week preoperatively and again postoperatively (before hospital discharge and at 3-month follow-up if they had deficits at discharge) on a neuropsychological battery. Different forms of each test were used preoperatively and postoperatively to reduce practice effects. Paired t tests were used to examine differences between preoperative and postoperative test scores across individuals. RESULTS: On most tests, there was no significant change between preoperative and postoperative scores. A significant decline in accuracy before hospital discharge was found only in figure copying (P<0.04) and associative learning (P<0.01), and significant slowing was found on 1 test (P<0.01). Even on these tests, only 3 of 25 patients showed significant deterioration. All but 1 patient returned to baseline by the 3 month follow-up. CONCLUSIONS: We found no evidence of subtle cognitive deficits resulting from aneurysm clipping alone, suggesting that the common impairments after surgery for ruptured aneurysms are due to SAH itself, complications of SAH such as vasospasm or hydrocephalus, or perioperative stroke. PMID- 12907821 TI - Hyperglycemia exacerbates brain edema and perihematomal cell death after intracerebral hemorrhage. AB - BACKGROUND AND PURPOSE: Hyperglycemia has a deleterious effect on brain ischemia. However, the effect of hyperglycemia in intracerebral hemorrhage (ICH) is not well known. We investigated the effect of hyperglycemia on the development of brain edema and perihematomal cell death in ICH. METHODS: Hyperglycemia was induced by intraperitoneal injection of streptozotocin (60 mg/kg) in adult Sprague-Dawley male rats. ICH was induced by stereotaxic infusion of 0.23 U of collagenase into the left striatum. Seventy-two hours after ICH, terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) staining was performed for perihematomal cell death. We also measured brain water content to evaluate edema formation. RESULTS: The serum glucose level of the hyperglycemic group was 394.0+/-180.3 mg/dL (n=31), and that of the normoglycemic group was 97.5+/-27.4 mg/dL (n=31). The size of hemorrhage was similar between groups, without any significant difference (n=8 in each group). The brain water content of hyperglycemic rats (n=17) increased in both lesioned (81.0+/-0.5%) and nonlesioned hemispheres (78.7+/-0.6%) compared with the normoglycemic group (n=17; lesioned: 78.9+/-0.8%; nonlesioned: 77.3+/-1.1%). In the hyperglycemic group, more TUNEL-positive cells were found in the perihematomal regions (n=6). CONCLUSIONS: Hyperglycemia caused more profound brain edema and perihematomal cell death in experimental ICH. PMID- 12907822 TI - Profiles of matrix metalloproteinases, their inhibitors, and laminin in stroke patients: influence of different therapies. AB - BACKGROUND AND PURPOSE: The goal of this study was to determine the temporal profile of several matrix metalloproteinases (MMPs), tissue inhibitors of MMPs (TIMPs), and laminin (an MMP substrate) in human stroke under different treatment paradigms, including thrombolysis and hypothermia. METHODS: We serially measured the serum levels of MMP-2, MMP-3, MMP-9, MMP-13, TIMP-1, TIMP-2, and laminin in 50 patients with acute ischemic stroke using zymography or enzyme-linked immunosorbent assay. Patients were treated with heparin, therapeutic thrombolysis, or hypothermia. Scandinavian Stroke Scale scores were obtained at baseline. Infarct volume was measured with CT scanning on day 4 after stroke onset. Healthy persons were used as control subjects. RESULTS: MMP-2 and MMP-9 increased during the course of ischemia, whereas intact laminin and TIMP-2 decreased significantly (P<0.05). MMP-9 and laminin levels varied significantly by infarct size (P=0.001) and therapy (P=0.0005). MMP-9 levels were significantly higher in patients treated with tissue plasminogen activator (tPA) compared with patients treated with hypothermia. The cleaved form of MMP-9 was found solely in 4 patients treated with tPA. Intact laminin levels were significantly lower in the tPA group than in the hypothermia group. CONCLUSIONS: Selected MMPs and TIMPs are involved in the pathophysiology of acute stroke. This is also reflected by changes in laminin. Treatment paradigms differentially influence levels of MMP-9 and laminin. Combination therapies explicitly involving MMP inhibition could be of value in future treatment strategies. PMID- 12907823 TI - Stroke in South America: a systematic review of incidence, prevalence, and stroke subtypes. AB - BACKGROUND AND PURPOSE: Stroke is a leading cause of mortality and disability in South America because of an increase in life expectancy and changes in the lifestyle of the population. Because epidemiological and clinical characteristics of stroke vary according to regional factors, we need to know the peculiarities of stroke on this continent. METHODS: We performed a systematic review of articles on stroke in South America, with emphasis on those providing information on the incidence and prevalence of stroke (community-based studies) and the pattern of stroke subtypes (hospital-based studies). RESULTS: Seven papers provided information on stroke epidemiology; 11 gave data on the pattern of stroke subtypes. Community-based studies showed crude stroke prevalence rates ranging from 1.74 to 6.51 per 1000 and annual incidence rates from 0.35 to 1.83 per 1000. Hospital-based stroke registries consistently reported a high frequency of intracranial hemorrhages, which accounted for 26% to 46% of all strokes. Among patients with cerebral infarctions, intracranial atherosclerotic lesions and small-vessel disease have been common pathogenic mechanisms underlying the stroke. In most studies, hypertensive arteriolopathy was the most common cause of both infarctions and hemorrhages. CONCLUSIONS: Stroke has been poorly studied in South America. Available data suggest that the prevalence and incidence of stroke are lower than in developing countries. The pattern of stroke subtypes seems to be different from that reported in other regions of the world, with a higher frequency of cerebral hemorrhages, small-vessel disease, and intracranial atherosclerotic lesions. Such differences may be related to genetic, environmental, or sociocultural factors and to differences in the control of stroke risk factors. PMID- 12907824 TI - Iron deficiency: causes, consequences, and strategies to overcome this nutritional problem. AB - Iron deficiency and anemia affect a substantial portion of the world's population, provoking severe health problems to the people suffering these conditions, as well as important economic losses to the regions in which this nutritional deficiency is significant. In this work, the principal causes and consequences produced by this deficiency are discussed, as well as the different strategies that can be applied in order to prevent and solve this nutritional problem. PMID- 12907825 TI - Serum zinc in healthy Belgian children. AB - Many reports mention marginal zinc status in childhood. Information on serum zinc (Zn) in Belgian children since the last reports are old and feeding habits are changing. Four hundred fifty-seven healthy children (0-14 yr, 262 boys) had a venipuncture after an overnight fast during a vaccination campaign. Serum Zn, alpha-tocopherol (alpha-T), cholesterol, low-density lipoprotein cholesterol (LDL C), high-density lipoprotein cholesterol (HDL-C), apolipoprotein B (Apo B), Apo A, and malondialdehyde (MDA) were determinated. The median Zn value is lower in infants than in older children (respectively 11.6 micromol/L vs 12.8 micromol/L). The type of infant feeding does not influence the serum Zn concentrations (breast feeding, adapted, hypoallergenic, soy, or thickened). No children had increased serum MDA concentrations and the value is not influenced by the Zn concentration. Children presenting higher serum Zn values also have significantly higher serum alpha-T levels. In infants, there is a significant positive correlation between serum Zn and cholesterol, LDL-C, and Apo B. In this apparently healthy population, no signs of abnormal in vivo peroxidation of fatty acids are observed, even in the children with low serum Zn. More sensitive methods for the detection of peroxidation are necessary for determination of in vivo effects of marginal trace element status. PMID- 12907826 TI - Should chronic obstructive pulmonary disease outpatients be routinely evaluated for trace elements? AB - We searched for serum concentrations of trace elements and correlated them to malondialdehyde (MDA), which is an indirect marker of oxidative stress, in order to clarify if routine evaluation is necessary in chronic obstructive pulmonary disease (COPD) outpatients. Serum concentrations of copper (Cu), zinc (Zn), and magnesium (Mg) were determined by atomic absorption spectrophotometry and iron (Fe) by a ILLab 1800 autoanalyzer with ILLab test kits. Serum MDA concentrations were detected in terms of TBARS (thiobarbituric acid reactive substances) spectrophotometrically. Serum Cu, Zn, Mg, Fe, and MDA concentrations in patient and control groups were all in the normal reference range. The results respectively were as follows: Cu:123 +/- 29.2 and 122.2 +/- 23.4 microg/dL; Zn: 87.8 +/- 17.8 and 96.9 +/- 12.9 microg/dL; Mg: 2.3 +/- 0,5 and 2.04 +/- 0.28 mg/dL; Fe: 73.8 +/-35.5 and 80.7+/-51.2 microg/dL; MDA: 1.09+/-0.11 and 0.95+/ 0.06 nmol/L. MDA was not correlated to Cu, Zn, Mg, or Fe (p>0.05 for all). The serum Zn concentration of COPD group was lower than the control group (p=0.042), whereas the Mg concentration was higher (p=0.021). There was no statistical difference in other study parameters. Oxidative stress was not increased in clinically stable, regularly treated COPD patients. Although there was no deficiency in trace elements (Cu, Fe, Mg, and Zn), serum Zn was close to the lower limit of the reference value. There is no need for routine evaluation of trace elements in clinically stable, regularly treated COPD outpatients. PMID- 12907827 TI - Zinc-induced changes in ionic currents of cardiomyocytes. AB - The whole-cell voltage-clamp technique was applied to isolated ventricular myocytes to investigate the effects of extracellular and intracellular zinc application on L-type Ca2+ channel currents (ICa). Extracellular zinc exposure at micromolar concentration induced a reversible (with washout of ZnCl2) reduction (30%) of ICa with no change in current-voltage relationship. On the other hand, an increase of intracellular free-zinc concentration, [Zn2+]i, from normal (less than 1 nM) to approx 7 nM with 10 microM Zn-pyrithione exposure caused an inhibition of 33+/-6% in the peak of the ICa and altered the voltage dependency of L-type Ca2+ channels with a 10-mV left shift and a hump at around -40 mV in its current-voltage relation. In contrast, N,N,N',N'-tetrakis(2 pyridylmethyl)ethylenediamine (TPEN) strongly inhibited the ICa (42+/-2%), with only a small but detectable outward shift of the holding current measured at the end of the pulses. Zn-pyrithione and TPEN caused a reproducible decrease of the ICa. Interestingly, TPEN application, without Zn-pyrithione pretreatment, inhibited the ICa (35+/-2%) with no change in voltage dependency. Taken together, the results suggest that both extracellular and intracellular zinc increases under pathological conditions in cardiomyocytes can alter the ICa, but their effects are not in the same order and same manner. One should consider these possible side effects when it is suggested to be vital to cardiovascular cell integrity and functions. PMID- 12907828 TI - Enhanced testicular antioxidant capacity in streptozotocin-induced diabetic rats: protective role of vitamins C and E and selenium. AB - Diabetes mellitus is associated with diabetic impairment of testicular function, ultimately leading to reduced fertility. Its etiology may involve oxidative damage by reactive oxygen substances, and protection against this damage can be offered by antioxidant supplementation. The aim of this study was to investigate the effects of intraperitoneal administration of vitamin C and E, selenium (Se), and vitamin E plus Se (COM) on concentrations of lipid peroxide (as malondialdehyde; MDA), reduced glutathione (GSH), and vitamin E concentrations and glutathione peroxidase (GSH-Px) activity in the testes of rats with diabetes induced by streptozotocin (STZ). Sixty groups were used (10 animals each) and these animals were initially allocated to two groups: control group and diabetic group. The diabetic group was subdivided into five groups as follows: diabetic control (DC), vitamin E, Se, COM, and vitamin C. Animals in the DC group and vitamin C, vitamin E, Se, and COM groups were made diabetic by the injection of STZ on 4 d after an injection of vitamins C and E, Se, and COM. Those vitamins and Se were also administered for 21 consecutive days. The MDA, vitamin E, GSH levels, and GSH-Px activities in testes were determined. Although the vitamin E concentration was higher in the control than in the DC group, the MDA levels were found to be lower in the control than in the DC group. The MDA levels in the testes samples of vitamin C, vitamin E, Se, and COM groups were lower than the DC group. However, GSH-Px activity and GSH levels in the testes were not significantly different between the control and DC groups. Vitamin E concentrations in the vitamin C, vitamin E, Se, and COM groups and GSH levels and GSH-Px activities in the Se, COM, and vitamin C groups were higher than either the control or DC group. The results indicate that reactive oxygen substances may be involved in possible testicular complications in diabetes of rats. Administration of vitamins C and E and Se reduced the testicular lipid peroxidation; these vitamins and Se had significant protective effects on testes of rats against oxidative damage in diabetes. PMID- 12907829 TI - Stabilized ferrous gluconate as iron source for food fortification: bioavailability and toxicity studies in rats. AB - The iron bioavailability and acute oral toxicity in rats of a ferrous gluconate compound stabilized with glycine (SFG), designed for food fortification, was studied in this work by means of the prophylactic method and the Wilcoxon method, respectively. For the former studies, SFG was homogeneously added to a basal diet of low iron content, reaching a final iron concentration of 20.1 +/- 2.4 mg Fe/kg diet. A reference standard diet using ferrous sulfate as an iron-fortifying source (19.0 +/- 2.1 mg Fe/kg diet) and a control diet without iron additions (9.3 +/- 1.4 mg Fe/kg diet) were prepared in the laboratory in a similar way. These diets were administered to three different groups of weaning rats during 23 d as the only type of solid nourishment. The iron bioavailability of SFG was calculated as the relationship between the mass of iron incorporated into hemoglobin during the treatment and the total iron intake per animal. This parameter resulted in 36.6 +/- 6.2% for SFG, whereas a value of 35.4 +/- 8.0% was obtained for ferrous sulfate. The acute toxicological studies were performed in two groups of 70 female and 70 male Sprague-Dawley rats that were administered increasing doses of iron from SFG. The LD50 values of 1775 and 1831 mg SFG/kg body wt were obtained for female and male rats, respectively, evidencing that SFG can be considered as a safe compound from a toxicological point of view. PMID- 12907831 TI - Oral iron absorption test: should it be performed before starting treatment with ferrous preparations? AB - The oral iron absorption test is sometimes used in the assessment of ferrous preparation efficacy before therapeutic use in the treatment of patients with anemia. Overdoses of Fe can cause the production of free radicals that are dangerous because of chemical modifications and damage of proteins, lipids, carbohydrates, and nucleotides. We suggest that this test should not be performed with the recommended dose of iron because of the potential threat to the patients. We assessed the serum concentration of iron and total iron-binding capacity during the test. Before and after the test, the concentration of malonyldialdehyde, an end product of lipid peroxidation, was determined in serum. In most patients, we found an increase in malonyldialdehyde concentration, suggesting the enhanced production of free radicals. This increase was particularly marked in patients with an overabsorption of iron. The administration of iron in the dose recommended for the oral iron absorption test causes increase in serum malonyldialdehyde concentration, proving an overproduction of free radicals. This test should not be performed because of the evidence proving detrimental effects of free-radical overproduction on the human body. PMID- 12907830 TI - Effects of iron(II) salts and iron(III) complexes on trace element status in children with iron-deficiency anemia. AB - Iron-deficiency anemia (IDA) is the most common nutritional deficiency in childhood throughout the world. Although it has been shown that IRA is associated with elevated plasma copper and depleted zinc levels in children, there are conflicting results on the effect of iron supplementation on the absorption of these elements. The aim of this study was to investigate the effects of ferrous and ferric iron supplementation on the trace element status in children (n=25, aged 8-168 mo) with IDA. Fourteen of them were treated with ferric hydroxide polymaltose complex (Ferrum, Vifor, Switzerland) (6 mg/d in the first 3 mo for initial therapy and 3 mg/kg for 3 mo as maintenance); the others were treated with a ferrous sulfate complex (FerroSanol, Schwarz, Germany) (6 mg/d in the first 3 mo for initial therapy and 3 mg/kg for 3 mo as maintenance). Plasma copper, zinc, and ceruloplasmin levels as well as hematological parameters were determined at baseline and the first, third, and sixth month of the treatment period. The hemoglobin and iron levels of patients in both groups were higher in the first and sixth months compared to baseline. Although the ceruloplasmin levels were depleted (48.9 mg/dL vs 41.4 mg/dL, p=0.035) during ferrous iron treatment, the copper and zinc levels remained unchanged. On the other hand, ferric iron supplementation led to an increase in zinc levels in the sixth month of treatment (0.77 mg/L vs 1.0 mg/L, p=0.021). The plasma copper levels were lower in the ferrous iron-treated group at the end of the first month of treatment than in the ferric irontreated group (1.06 mg/L vs 1.29 mg/L, p=0.008). In conclusion, our data showed that copper and ceruloplasmin metabolisms were affected by ferrous iron supplementation, whereas ferric iron kept them to normal levels of zinc, possibly by affecting their absorption. We conclude that the copper and zinc status of patients with IDA should be taken into consideration before and after iron therapy. PMID- 12907832 TI - Topical vaccination of DNA antigens: topical delivery of DNA antigens. AB - Topical DNA vaccines have been shown to elicit both broad humoral and cellular immune responses in vivo. The skin is an attractive site for the delivery of DNA antigens for DNA vaccination. However, due to skin's barrier properties, the penetration of DNA and the applications of topical vaccination are limited. To improve permeability, chemical and physical approaches have been examined to decrease stratum corneum barrier properties. Topical vaccination has been achieved using topical application of naked DNA, DNA/liposomes or emulsion complex, liposomal cream, as well as physical methods such as stripping, electroporation, and micromechanical disruption methods. All methods resulted in a significant enhancement in humoral and cellular immune responses over naked DNA alone. To develop more cost-effective and needle-free vaccines, skin-targeted immunizations are required. This review focuses on the chemical and physical methods developed to enhance DNA delivery into skin. PMID- 12907833 TI - Impact of ultraviolet radiation and ozone on the transepidermal water loss as a function of skin temperature in hairless mice. AB - Exposure to ultraviolet radiation or ozone leads to skin damage including oxidation of skin biomolecules, as well as to depletion of constitutive antioxidants. The highly organized stratum corneum forming the main barrier against most xenobiotics is particularly susceptible to such damage and possible barrier perturbation may be the consequence. Whereas ample evidence exists for an increased permeability for different solutes including water after exposure to ultraviolet radiation, such an effect has not yet been reported for ozone. This study reports on the effect of such oxidative stressors using the hairless mouse as the skin model and measuring temperature-controlled transepidermal water loss (TEWL) as an indicator for skin barrier integrity. First, a strong dependency of the TEWL on skin temperature was observed, an effect that was clearly more pronounced than that found in man. Given this temperature dependency in untreated animals, we proceeded to determine the effects of both ultraviolet radiation and ozone on TEWL over a relevant physiological skin temperature range. Solar simulated ultraviolet radiation (0.75-3 minimal erythemal dose) resulted in a delayed and dose-dependent skin barrier disruption over the entire temperature range investigated. Conversely, daily ozone exposure at 2 ppm for 1 week, however, did not significantly alter TEWL up to 72 h after the last exposure. The results demonstrate a differential response of the epidermis to two environmental stressors associated with oxidative damage; they suggest that chronic ozone exposure at relevant environmental levels does not lead to a detectable skin barrier defect, while solar UV exposure was demonstrated to increase epidermal water loss. Furthermore, experimental evidence clearly suggests that future studies applying TEWL measurements in animal models should be performed under carefully controlled skin temperature conditions. PMID- 12907834 TI - Mass spectrometric examinations of stratum corneum lipid models exposed to ultraviolet irradiation. AB - Lipid model systems consisting of the major components of the stratum corneum intercellular lipid matrix were studied to investigate the ultraviolet-radiation mediated damage of these biomolecules. Pure lipids and liposomes were irradiated using a lamp emitting a solar radiation spectrum. The influences of the irradiation and the effects of added iron ions were studied by electrospray ionization mass spectrometry (MS) with an ion trap analyser. Exact mass measurements were carried out using a time-of-flight mass spectrometer. Only linolenic acid and cholesterol were found to be subject to oxidative changes caused by UV irradiation whereas the other lipids examined (dipalmitoylphosphatidylcholine, ceramide III and cholesterol sulphate) were stable to oxidative stress. Several lipid adducts were observed upon analysis of the liposomes. The composition of these adducts was identified by MS/MS experiments. PMID- 12907835 TI - High plasma levels of 8-methoxypsoralen following bath water delivery in dermatological patients. AB - With respect to the clinical advantages known for bath PUVA therapy, it was of interest to compare the plasma levels of 8-methoxypsoralen (8-MOP) in bath therapy with those after oral administration for a better insight into the pharmacokinetics of 8-MOP following different modes of application. Considerable high plasma levels of 8-MOP were observed after bath therapy with interindividual variability. The half-life of plasma 8-MOP was markedly shorter after bath PUVA than after oral application. The pharmacokinetic profile of 8-MOP differs according to the mode of application. PMID- 12907836 TI - Relationship of hairless mouse skin surface temperature to wound severity and maturation time. AB - Skin surface temperature (SST) changes measured on live hairless mice are presented as a simple means of following wound healing. SST is generally determined by 3 factors which are the ambient temperature, the rate of water loss from the surface of the skin and the diffusion of thermal energy from the body's core. The SST increase immediately after a burn injury reflects the amount of thermal energy absorbed by the skin surface. For burns and other injuries, the drop in SST following injury, but after thermal equilibrium has been established, provides an early indication of the degree of impairment of the skin's barrier. Three kinds of mouse skin wounds, mechanical (tape stripping), thermal and chemical (phenol application), were investigated. SST nadirs ranged from 2.1 to 4.4 degrees C with mild to full-thickness burns, respectively. Except for the earliest moments after wounding, striking parallels were noted between SST and transepidermal water loss profiles for these injuries. The SST profile over the full course of wound healing clearly indicates the severity of the injury, the stages of wound maturation and the time to complete skin healing. PMID- 12907837 TI - EEMCO guidance for the efficacy assessment of antiperspirants and deodorants. AB - Overproduction of sweat, sweaty skin and body odours are unpleasant for many social groups. Body cleansing products are designed to combat these undesirable features of skin. In addition, antiperspirant and deodorant products are more specifically used in the underarm site by a large part of the adult population. Antiperspirants are offered to control emotionally triggered sweating in the armpit. Deodorants are designed to combat malodour generated from bacteria modified sweat. This review summarizes the physiology of eccrine, apocrine and apoeccrine sweat glands. The mechanisms of action of antiperspirants and deodorants are described as well as the factors influencing their efficacies. A series of tests using various measurement methods can be used to demonstrate the efficacy of antiperspirants. These include the gravimetric method, water evaporation quantification, electrodermal measurements, staining procedures, dye injections and cyanoacrylate skin surface strippings and casting replicas. Deodorant efficacy can be evaluated by sensory assessments performed by an expert panel. Indirect support is provided by visualization of apocrine gland excretion and collection of sweat and volatile compounds. Microbiological assessments and chromatographic analysis also provide indirect information. PMID- 12907838 TI - Entry of blood-borne cytokines into the central nervous system: effects on cognitive processes. AB - Blood-borne cytokines affect many aspects of the central nervous system (CNS). One of the more dramatic effects is the induction of sickness behavior. Impairments in learning and memory are an important component of sickness behavior and are largely mediated by IL-1. Many mechanisms have been proposed by which a cytokine circulating in the blood can affect functions within the CNS. We review here the role one of those mechanisms, that of transport of cytokines across the blood-brain barrier (BBB), plays in induction of the memory impairments of sickness behavior. We have shown that the posterior division of the septum (PDS) plays a key role in mediating the effects of interleukin-1alpha (IL-1alpha) on memory. Furthermore, this effect at the PDS is largely mediated by circulating IL-1alpha acting directly at the PDS which, in turn, depends on the ability of IL-1alpha to cross the BBB. PMID- 12907839 TI - Down-regulation of Bcl-2, activation of caspases, and involvement of reactive oxygen species in 6-hydroxydopamine-induced thymocyte apoptosis. AB - OBJECTIVE: Our previous work showed that 6-hydroxydopamine (6-OHDA) induced mouse thymocytes to undergo apoptosis both in vivo and in vitro. In the present study, we further investigated the mechanisms of 6-OHDA-induced apoptosis in vitro. METHODS: Naive mouse thymocytes were cultured with 6-OHDA. The percentages of apoptotic cells were quantified by propidium iodide staining, and DNA fragmentation was detected by agarose gel electrophoresis. Intracellular Bcl-2 was analyzed by immunofluorescence staining. Cu/Zn superoxide dismutase (Cu/Zn SOD) activities were measured by the SOD-525 method. RESULTS: The apoptotic effect of 6-OHDA was blocked by desipramine, a catecholamine uptake blocker. Treatment with 6-OHDA caused a reduction in Bcl-2 expression. VAD-FMK, a broad spectrum caspase inhibitor, and DEVD-CHO, a potent inhibitor of caspase-3, could block 6-OHDA-induced thymocyte apoptosis. However, the specific caspase-1 (ICE) inhibitor YVAD-CMK had no effect. This cell death process was prevented by the protein synthesis inhibitor cycloheximide and by antioxidants. The level of Cu/Zn SOD activities also decreased after cells were exposed to 6-OHDA. CONCLUSION: These results suggest an apoptotic effect of 6-OHDA via the uptake of this neurotoxin by thymocytes, and that down-regulation of Bcl-2, activation of caspases, such as caspase-3 but not caspase-1, generation of reactive oxygen species, and new synthesis of proteins are involved in this process. PMID- 12907840 TI - Coemergence of insomnia and a shift in the Th1/Th2 balance toward Th2 dominance. AB - OBJECTIVES: Insomnia is associated with physical and mental disorders. We examined the effect of insomnia on immune functions, focusing on the T helper 1 (Th1)/ T helper 2 (Th2) balance, by a cross-sectional design. METHODS: We provided a self-administered questionnaire to evaluate sleep habits, smoking and medical disorders to 578 men without any toxic exposure (20-64 years old), and measured natural killer (NK) cell activity in 324 men and production of interferon-gamma (IFN-gamma) and interleukin-4 (IL-4) after stimulation with phytohemagglutinin in 254 men. According to the criteria of DSM-IV, in which insomnia is classified into primary and secondary insomnia, we assessed the effect of insomnia on immune functions, controlling for age and smoking in groups with and without medical disorders. RESULTS: The prevalence of insomnia in the present study was 9.2%. In the absence of medical disorders, insomniac men had a significantly lower IFN-gamma and ratio of IFN-gamma to IL-4 than noninsomniac men. Men with insufficient sleep or difficulty initiating sleep (DIS) had a significantly lower IFN-gamma to IL-4 ratio than those not suffering from insufficient sleep or DIS. In the presence of medical disorders, insomniac men had significantly higher IL-4 than noninsomniac men. Men with difficulty maintaining sleep (DMS) had a significantly lower IFN-gamma to IL-4 ratio than men without DMS. NK cell activity was independent of insomnia. CONCLUSIONS: The present results showed a link between insomnia unrelated to medical disorders and a shift in the Th1/Th2 balance toward Th2 dominance, indicating that the relationship between sleep quality and the etiology of immune-related diseases should be reconsidered. PMID- 12907841 TI - Effect of interleukin 1beta on the HPA axis in H1-receptor knockout mice. AB - OBJECTIVES AND METHODS: Circulating cytokines such as interleukin-1 (IL-1), and tumor necrosis factor-alpha as well as lipopolysaccharide (LPS) are potent ACTH secretagogues, acting via stimulation of corticotropin-releasing hormone (CRH) and vasopressinergic neurons in the paraventricular nucleus of the hypothalamus (PVN). Histamine (HA) has been shown to stimulate ACTH secretion in rats, an effect in part mediated by CRH and arginine vasopressin (AVP). We have previously shown that inhibition of neuronal HA synthesis or central blockade of H(1) receptors (H(1)R) decreased the ACTH response to LPS in male rats. To further elucidate the role of neuronal HA in cytokine-induced activation of the HPA axis, we compared the effect of H(1)R knockout on IL-1beta-induced ACTH secretion in adult male mice. RESULTS: In H(1)R knockout mice, ACTH secretion increased from basal levels of 261 to 492 pmol/l in response to IL-1beta whereas the cytokine induced ACTH secretion increased from 140 to 406 pmol/l in wild-type mice. Plasma corticosterone (CORT) rose from basal levels of 99 to 831 nmol/l in knockout mice upon IL-1beta stimulation, whereas in wild-type mice CORT levels rose from 112 to 841 nmol/l. There was no significant difference in IL-1beta-stimulated plasma ACTH or CORT levels between wild-type and knockout mice. Furthermore, there was no significant difference in basal or IL-1beta-stimulated hypothalamic levels of histamine and tele-methyl-histamine between wild-type and knockout mice. HDC gene expression was significantly lower in knockout mice than in wild-type mice both under basal and IL-1beta-stimulated conditions, while there were no significant differences in CRH gene expression in the PVN in knockout mice under basal and IL 1beta-stimulated conditions. Increased basal expression of AVP in the PVN of knockout mice was observed in this study compared to wild-type mice. CONCLUSION: We conclude that the lack of the gene for histamine H(1)R does not seem to be crucial for the ACTH and CORT response to IL-1beta, either due to possible functional compensation in the H(1)R knockout mouse or due to activation of pathways other than the neuronal histaminergic system. PMID- 12907842 TI - Alzheimer's disease patients display gender dimorphism in circulating anorectic adipokines. AB - Among neurodegenerative diseases, Alzheimer's disease (AD) is a leading cause of death in elderly individuals. AD is characterized, among other clinical findings, by unexplained weight loss, cachexia and altered immune function. To explore whether any relationship between gender and circulating levels of several eating controlling metabolites exist, we evaluated leptin, tumor necrosis factor (TNF) alpha, triiodothyronine (T(3)), free (F) thyroxine (T(4)), TSH, PRL, insulin (INS), and cortisol in 15 AD-treated patients (age range 55-82 years): 9 postmenopausal females (without hormone replacement therapy) and 6 males. The results (mean +/- SEM) indicated that circulating leptin levels were significantly (p < 0.05) higher in female AD (40.34 +/- 11.1 ng/ml) than in male AD (6.07 +/- 1.39 ng/ml) patients. The difference found in circulating leptin levels was noticed regardless of BMI (26.75 +/- 1.77 and 24.55 +/- 1.93 kg/m(2), in females and males, respectively) and waist:hip ratios (0.91 +/- 0.03 and 0.94 +/- 0.02, in females and males, respectively). Moreover, serum TNF-alpha concentrations were also significantly (p < 0.02) higher in AD females (12.24 +/- 1.47 pg/ml) than in AD males (6.62 +/- 1.44 pg/ml), regardless of TNF-alpha:BMI ratios (0.50 +/- 0.09 and 0.28 +/- 0.08, in females and males, respectively; p > 0.05). Finally, no differences were observed between gender (in female and male AD patients, respectively) in circulating levels of T(3) (151.33 +/- 9.91 vs. 116 +/- 17.04 ng/dl), FT(4) (1.26 +/- 0.08 vs. 1.24 +/- 0.06 ng/dl), TSH (1.28 +/- 0.16 vs. 2.46 +/- 0.67 microIU/ml), PRL (10.53 +/- 2.47 vs. 12.61 +/- 2.37 ng/ml), INS (11.76 +/- 1.95 vs. 8.59 +/- 1.34 microIU/ml) and cortisol (15.71 +/- 1.23 vs. 12.63 +/- 1.47 microg/dl). These results indicate that our AD group of patients, with normal corticoadrenal and thyroid functions and normoprolactinemia, displayed a gender-related characteristic in the circulating levels of two very important anorectic signals, leptin and TNF-alpha, being both higher in female than in male AD patients, regardless of BMI. Our study suggests that increased circulating levels of both anorexigenic adipokines may contribute to the metabolic changes observed in AD females. PMID- 12907844 TI - Inhibition of constitutive nitric oxide production increases the severity of lipopolysaccharide-induced sickness behaviour: a role for TNF-alpha. AB - Administration of bacterial lipopolysaccharide (LPS) to rodents induces hypophagia, body weight loss and hypolocomotion, a constellation of symptoms collectively referred to as 'sickness behaviour'. We examined the role of the gaseous transmitter nitric oxide (NO) in mediating LPS-induced sickness behaviour in rats. Treatment with the non-selective NO synthase (NOS) inhibitor N(G)-nitro L-arginine (L-NA) (20 mg/kg; i.p.) increased the severity of LPS-induced sickness behaviour in rats, suggesting that endogenous NO does not act as a mediator of LPS-induced sickness behaviour, but may rather have a protective role, acting in an inhibitory feedback manner to limit LPS-induced sickness. To evaluate the role of the different NOS isoforms in this response, we examined the effect of the neuronal NOS inhibitor, 7-nitroindazole (7-NI; 25 and 50 mg/kg; i.p.), and the inducible NOS inhibitor, aminoguanidine (AGN; 50 and 100 mg/kg; i.p.). Neither 7 NI nor AGN significantly altered LPS-induced sickness behaviour. Therefore, it is likely that the endothelial isoform of NOS mediates the effect of L-NA on LPS induced sickness behaviour. As pro-inflammatory cytokines are mediators of LPS induced sickness behaviour, we examined the effect of L-NA (20 mg/kg; i.p.) on LPS-induced interleukin (IL)-1beta, IL-6 and tumour necrosis factor (TNF)-alpha production. L-NA increased LPS-induced TNF-alpha without significantly altering IL-1beta or IL-6 production. Moreover, pre-treatment with the TNF-alpha inhibitor pentoxyfilline (25 mg/kg; i.p.) largely reversed the augmenting effect of L-NA on LPS-induced sickness behaviour, suggesting that the ability of L-NA to increase TNF-alpha production underpinned its ability to increase the severity of sickness. In conclusion, L-NA increases the severity of LPS-induced sickness behaviour, most likely by blocking the tonic inhibitory action of constitutively produced NO on TNF-alpha production. PMID- 12907845 TI - Cerebral vascular responses to changes in carbon dioxide tension in term and preterm infants with apnea. AB - Carbon dioxide (CO(2)) plays important roles in regulating both respiratory drive and cerebral blood flow. These effects are mediated, in part, by activity of the sympathetic nervous system. We hypothesized that the presence of acute life threatening events or apnea in term or preterm infants, respectively, would serve as a marker for immaturity of cerebral autonomic innervation and that such infants would display a reduced cerebral vascular response to elevated pCO(2). Therefore, we evaluated the cerebral vascular response during CO(2) challenge tests in groups of term and preterm infants with primary apnea. In term infants (39 +/- 2 weeks gestation) with acute life-threatening events, elevated pCO(2) was accompanied by decreasing pulsatility index and increasing mean anterior cerebral blood flow velocity. However, in preterm infants (29 +/- 2 weeks' gestation) with apnea, pulsatility index and anterior cerebral artery flow velocity did not significantly change in response to CO(2) supplementation. We conclude that preterm, but not term, infants with apnea exhibit impaired vascular responses to hypercarbia. PMID- 12907843 TI - Chronic mild stress in mice decreases peripheral cytokine and increases central cytokine expression independently of IL-10 regulation of the cytokine network. AB - OBJECTIVES: Accumulating evidence indicates that stress leads to an increased expression of pro-inflammatory cytokines such as interleukin (IL)-6. The production and action of pro-inflammatory cytokines are down-regulated by anti inflammatory cytokines such as IL-10. This makes IL-10-deficient mice a potentially useful model to assess the effects of stress on cytokine production. METHODS: In the present study, IL-10-deficient mice were compared to wild-type mice in their behavioural and cytokine response to a chronic mild stress procedure. RESULTS: The 3-week chronic mild stress decreased body weight gain and sucrose consumption. It also resulted in a decreased expression of peripheral IL 1beta and IL-6 and an increased expression of brain IL-6. This last change in IL 6 was correlated to body weight loss in stressed mice. However, IL-10-deficient mice did not differ from wild-type mice in their response to the chronic mild stress procedure, despite substantial differences in functioning of the cytokine network. CONCLUSION: These results are interpreted in the context of the relationship between cytokines and behaviour. PMID- 12907846 TI - Lectin binding in the human umbilical cord from normally grown pregnancies and from pregnancies complicated by intra-uterine growth retardation with absent or reversed diastolic flow. AB - The content and the distribution of the glycoconjugate sugar residues were investigated in the human umbilical cord from normally grown pregnancies and from pregnancies complicated by intra-uterine growth retardation with absent or reversed flow in the umbilical artery. The gestational age range was 29-41 weeks. For this purpose, a battery of eight horseradish peroxidase conjugated lectins was used (ConA, WGA, PNA, SBA, DBA, LTA, UEA I, and OOA; see text). Our data showed that in both study groups ConA, WGA and PNA (after neuraminidase digestion) and LTA, UEA I, and OOA reacted with Wharton's jelly and vessels of the umbilical cord from the first weeks of gestation considered. SBA reactivity was observed only in the amniotic epithelial cells of the jelly and in the endothelial cells of the vessels. The reactivity with the most part of the lectins decreased or was lacking in the various components of the umbilical cord between 39 and 41 weeks of gestation in the normal group and between 36 and 37 weeks of gestation in the pathological one. Moreover, the intensity of reactivity with ConA, WGA and PNA (after neuraminidase digestion) and LTA, UEA I, and OOA in the normal group and with PNA (before and after neuraminidase digestion) and LTA and OOA in the pathological one was stronger in the peripheral portion of the jelly with respect to the adventitial one. The reactivity with ConA and UEA I was stronger in the vein than in the arteries in both study groups. Some hypotheses are reported on the role played by the sugar residues in the normal and pathological umbilical cords. PMID- 12907847 TI - Analysis of DNA damage using the comet assay in infants fed cow's milk. AB - It has been hypothesized that non-human milk feeding may increase the risk for cancer or for a specific cancer or group of cancers as well as the risk for diseases such as type-1 diabetes mellitus and Crohn's disease. Regarding DNA damage leading to cancer development in the absence of human milk protection, a comparison between infants fed human milk and cow's milk has been performed. Each group consisted of 35 infants, whose ages ranged from 9 to 12 months. The level of DNA damage in the peripheral blood lymphocytes of infants has been studied by the comet assay. A significant increase has been found in the number of limited DNA-damaged (p < 0.001) and extensive DNA-damaged (p < 0.001) cells of infants fed cow's milk. To our knowledge, this is the first study using the comet assay on infants not breast-fed. Supporting our previous SCE study, these results suggest that there is some level of DNA damage in the lymphocytes of infants not breast-fed and this may lead to malignancy in childhood or later in life. PMID- 12907848 TI - Sequential postnatal changes in erythrocyte glutathione and sulfhydryl content: a possible adaptational response to the extrauterine environment. AB - Glutathione is a key factor in intracellular antioxidant defense mechanisms. We postulated that it might modulate the adaptational response of the fetus to a higher oxygen environment after birth. Therefore, we determined total glutathione (GSH + GSSG) and non-protein sulfhydryl (-SH) content in cord blood RBC hemolysates, and sequential values in blood from the same infants for up to 72 h after birth. We noted a consistent rapid decline in total glutathione content by 24 h in both term (-18%) and preterm (-24%) infants, with subsequent stabilization of levels in term infants. However, erythrocyte (-SH) content (measured only in term babies) remained stable and unchanged from levels in cord blood, indicating the presence of (-SH) in one or more other moieties in addition to GSH. We speculate on the possibility of a role for GSH as a (-SH) donor in modulating postnatal fetal adaptation. PMID- 12907849 TI - Long-term effects of antepartum bed rest on offspring. AB - We studied the children of mothers who were confined to bed during pregnancy for more than 15 consecutive days (mean 3.69 months) in the years 1986-1990 (bed rest offspring; BRO). We studied 86 children: 43 BRO and 43 control children. Data were obtained by means of a 20-item questionnaire filled in by the mothers. The BRO group differed from the control group in incidence of allergies (p = 0.005), motion sickness (p = 0.03), and need to be rocked to fall asleep (p = 0.01). More BRO born at term than controls played musical instruments later in life. Two possible explanations for more allergies among the BRO group are the use of beta stimulating drugs against premature delivery and the effects of prolonged stress on the developing immune system. Understimulation of the developing vestibular system during gestation may affect some aspects of its development and may explain the higher incidence of motion sickness and need for vigorous rocking in BRO. PMID- 12907850 TI - Enteral crude red kidney bean (Phaseolus vulgaris) lectin--phytohemagglutinin- induces maturational changes in the enterocyte membrane proteins of suckling rats. AB - This study aimed to investigate the effect of enterally administered crude red kidney bean (Phaseolus vulgaris) lectin, PHA, on the expression of brush-border membrane vesicle (BBMV) proteins, in particular Na(+)/H(+) exchangers (NHEs), in the small intestine of suckling rats. Gavage of PHA to 14-day-old rats for 3 days resulted in altered protein/glycoprotein patterns as analyzed by SDS-PAGE. Immunoblots demonstrated the appearance of two 71- and 27-kD protein bands indicative for NHE3--one of the NHE isoforms--and PHA, respectively. PHA treatment also resulted in an augmented uptake of (22)Na(+) by the BBMV indicating an increase in NHE activity. Overall, the data suggests that enteral PHA exposure may induce maturational changes in enterocyte membrane proteins in young rats. In view of these findings, an investigation into the addition of PHA to infant formulas and weaning diets is warranted. PMID- 12907851 TI - Alterations in cerebral mitochondria during acute hypoglycemia. AB - To test the hypothesis that acute hypoglycemia leads to free radical induced alterations in cerebral mitochondria, newborn piglets were subjected to 2 h of insulin-induced hypoglycemia (blood glucose 1 mmol/l). The effects of free radicals were determined in cerebral cortical synaptosomes, mitochondria, and neuronal nuclei by measuring membrane lipid peroxidation. Fragmentation of nuclear and mitochondrial DNA was also examined. Lipid peroxidation was significantly increased in hypoglycemic mitochondrial membranes as compared to controls, but no increase in peroxidation in hypoglycemic synaptosomal or nuclear membranes was observed. An increase in low molecular weight DNA fragments was observed only in mitochondrial DNA from hypoglycemic piglets. We speculate that alteration of cerebral mitochondria due to increased free radical production is one of the early events in the pathogenesis of hypoglycemic brain injury. PMID- 12907852 TI - Postischemic hyperthermia induced caspase-3 activation in the newborn rat brain after hypoxia-ischemia and exacerbated the brain damage. AB - The effects of postischemic hyperthermia were investigated in the newborn rat brain after hypoxia-ischemia (HI). Seven-day-old rats were subjected to left carotid artery ligation followed by 8% oxygen for 30 min, and divided into a hyperthermia group (rectal temperature at 39 degrees C for 6 h) and a normothermia group. Hyperthermia resulted in an approximately 5-fold increase in activated caspase-3 24 h after HI when compared with the normothermia group, and gross loss of brain tissue was observed only in the hyperthermia group at 7 and 30 days after HI. Our results show that postischemic hyperthermia exacerbates HI injury in immature brains, and that the mechanism is strongly associated with activation of an apoptotic pathway. PMID- 12907853 TI - Vascular reactivity of pulmonary arteries from premature lambs subjected to liquid ventilation. AB - This study aimed to evaluate the effect of tidal liquid ventilation (TLV) and liquid assisted high frequency oscillatory ventilation (LA-HFOV) on the contractile and relaxing properties of fetal ovine pulmonary arteries. 0.85 term lambs were subjected, after surfactant instillation, to 5 h of TLV or LA-HFOV (5 ml.kg(-1) perfluorocarbon liquid). After euthanasia of the animals, intrapulmonary arteries (fourth branch) were dissected and mounted in a myograph for isometric tension recording. Arteries from unventilated age-matched fetuses were also studied. The contractions induced by K(+), and the thromboxane A(2) mimetic U46619 were not significantly different in the TLV and the LA-HFOV groups. Acetylcholine-induced relaxation was absent after TLV and LA-HFOV but present in unventilated animals. Sodium nitroprusside-induced relaxation was also similar after TLV and LA-HFOV, but reduced when the two groups were compared with unventilated fetuses. We conclude that after TLV and LA-HFOV the pulmonary arterial responses to receptor- and non receptor-mediated contraction and to endothelium-dependent and -independent relaxation were similar. PMID- 12907854 TI - Increased wave latency in auditory brainstem response to anemia in newborn and adult sheep. AB - The objective of the present study was to determine whether phlebotomy-induced anemia in newborn lambs and adult sheep results in prolonged auditory brainstem response (ABR) wave latencies, and if found, whether developmental differences exist between the two groups. Among newborn lambs, hemoglobin (Hb) levels were reduced from 100-120 to 30-50 g/l over a 3-day period. Over the next 2 days, serial red blood cell transfusions were administered to restore Hb to pre phlebotomy levels. In adult sheep, comparable levels of phlebotomy-induced anemia were achieved within a 24-hour period, after which Hb levels returned to pre phlebotomy levels. During the induction of anemia and its reversal, a significant (p < 0.05) increase in ABR latency followed by a return to pre-anemia ABR latencies that was most prominent in the peripheral auditory pathway was observed in both newborn and adult sheep as Hb levels fell below 50 g/l (p < 0.05). There was no difference between adults and newborns in ABR latency relative to the Hb level at which ABR wave latencies increased. We speculate that ABR wave latency prolongation might serve as an indicator of the need for red blood cell transfusion. PMID- 12907855 TI - Neurogenesis in the hippocampus of an adult marsupial. AB - In the adult eutherian brain, stem cells in the dentate gyrus continually divide throughout adult life and into old age producing new granule cells. However, it was not known whether this is also the case for marsupials. Previously, in fact, it was thought that marsupials did not have continued neurogenesis in the mature brain. Here we examined neurogenesis in the adult brain of a small marsupial, the fat-tailed dunnart, using 3H-thymidine to label newly generated cells. We showed that neurogenesis takes place in the adult dentate gyrus along its inner margin, as seen in eutherian mammals. Control animals had similar numbers of labeled cells 24 h and 1 month after 3H-thymidine injection. An enriched environment resulted in similar numbers of cells being generated as controls. However, there was a significant decline in the number of labeled cells one month later. Stress and old age resulted in significantly lower numbers of new cells being generated. In immunohistochemically treated control brains, 3H-thymidine-labeled cells at the early stage were sometimes GFAP positive, were not calbindin positive at either stage examined and at the later stage were PSA-NCAM positive. We hypothesize that, as seen in eutherian mammals, the new cells progressed from being GFAP positive at stem cell stage to PSA-NCAM positive during outgrowth of mossy fibers 1 month later, to calbindin positive when mature. It is possible that maturity of these cells was not reached by 1 month as marsupials have a slower metabolic rate and this species also undergoes daily periods of torpor. PMID- 12907856 TI - Brain-body size relations in grasshopper mice. AB - Cranial volumes were measured on museum specimens of two species of grasshopper mice, Onychomys leucogaster brevicaudus and Onychomys torridus pulcher. These were compared with body weights and body lengths, as recorded on the museum tags. On average, females were heavier and slightly longer than males, but they had smaller cranial volumes by about 3%. Statistically, the differences were marginally significant (with no difference in body length for one species), but the pattern of differences was the same in both species, and therefore probably real. This casts suspicion on the common notion that a larger body size requires a larger brain. It also questions the real meaning of encephalization quotients. PMID- 12907857 TI - Parvalbumin-, calbindin-, and calretinin-immunoreactive neurons in the prefrontal cortex of the owl monkey (Aotus trivirgatus): a standardized quantitative comparison with sensory and motor areas. AB - Recent studies have revealed regional variation in the density and distribution of inhibitory neurons in different cortical areas, which are thought to reflect area-specific specializations in cortical circuitry. However, there are as yet few standardized quantitative data regarding how the inhibitory circuitry in prefrontal cortex (PFC), which is thought to be involved in executive functions such as cognition, emotion and decision making, compares to that in other cortical areas. Here we used immunohistochemical techniques to determine the density and distribution of parvalbumin (PV)-, calbindin (CB)-, and calretinin (CR)-immunoreactive (ir) neurons and axon terminals in the dorsolateral and orbital PFC of the owl monkey (Aotus trivirgatus), and compared them directly with data obtained using the same techniques in 11 different visual, somatosensory and motor areas. We found marked differences in the density of PV ir, CB-ir, and CR-ir interneurons in several cortical areas. One hundred and twenty eight of all 234 possible between-area pair-wise comparisons were significantly different. The density of specific subpopulations of these cells also varied among cortical areas, as did the density of axon terminals. Comparison of PFC with other cortical areas revealed that 40 of all 66 possible statistical comparisons of the density of PV-ir, CB-ir, and CR-ir cells were significantly different. We also found evidence for heterogeneity in the pattern of labeling of PV-ir, CB-ir, and CR-ir cells and axon terminals between the dorsolateral and orbital subdivisions of PFC. These data are likely to reflect basic differences in interneuron circuitry, which are likely to influence inhibitory function in the cortex. PMID- 12907858 TI - Spatio-temporal distribution of microglia/macrophages during regeneration in the cerebellum of adult teleost fish, Apteronotus leptorhynchus: a quantitative analysis. AB - In contrast to mammals, adult teleost fish exhibit an enormous capacity to replace damaged neurons with newly generated ones after injuries in the central nervous system. In the present study, the role of microglia/macrophages, identified by tomato lectin binding, was examined in this process of neuronal regeneration in the corpus cerebelli of the teleost fish Apteronotus leptorhynchus. In the intact corpus cerebelli, or after short survival times following application of a mechanical lesion to this cerebellar subdivision, microglia/macrophages were virtually absent. Conversely, approximately 3 days after application of the lesion, the areal density of microglia/macrophages started to increase at and near the lesion site in the ipsilateral hemisphere, as well as in the contralateral hemisphere, and reached maximum levels at approximately 10 days post lesion. The density remained elevated until it reached background levels approximately one month after the injury. By comparing the time course of the appearance of microglia/macrophages with that of other regenerative events occurring within the first few weeks of wound healing in this model system, we hypothesize that one possible function of microglia/macrophages might be to remove debris of cells that have undergone apoptotic cell death at the lesion site. PMID- 12907859 TI - Expression of nitric oxide synthase in the preoptic-hypothalamo-hypophyseal system of the teleost Oreochromis niloticus. AB - In the present study, we have analyzed the expression of nitric oxide synthase (NOS) in the preoptic-hypothalamo-hypophyseal system of the teleost Oreochromis niloticus. The assay for enzyme activity demonstrated that a constitutive NOS activity is present both in soluble and particulate fractions of the homogenates of diencephalons. Western blot analysis using an antibody against the N-terminus of human nNOS revealed two bands both in the supernatant and in the pellet. One band co-migrates at approximately 150 kDa with that detected in the rat cerebellum homogenates and presumably corresponds to neuronal NOS (nNOS) of mammals. The additional band, which migrates at approximately 180 kDa, might be attributed to an alternatively spliced nNOS isoform. Using NADPH diaphorase (NADPHd) histochemistry in combination with NOS immunohistochemistry, nNOS expression has been detected in preoptic nuclei, hypophysiotrophic nuclei of the ventral hypothalamus, and the pituitary gland. Various degrees of dissociation of NADPHd activity and nNOS immunoreactivity have been detected that could be attributed to the expression of different subtypes of nNOS in the preoptic/hypothalamo/hypophysial system of tilapia. In this paper, we also investigated the colocalization of nNOS with arginine-vasotocin (AVT) by means of immunolabeling of consecutive sections. Results suggest that NO may be colocalized with AVT in a subpopulation of neurosecretory neurons. Present findings suggest that nitric oxide (NO) is implicated in the modulation of hormone release in teleosts in a similar way to mammals. PMID- 12907860 TI - Queen discrimination ability of ant workers (Camponotus japonicus) coincides with brain maturation. AB - Discrimination behavior is a fundamental feature of social insects, and the discrimination ability allows the individual to be integrated into the colony. To address the brain functions associated with this task, which underlies intraspecific communication, I examined the ability to discriminate queens in young workers of the ant Camponotus japonicus together with a histological analysis of their brains. Workers raised in a foster colony were tested with respect to their ability to discriminate the foster queen from an unfamiliar queen. During the first 3 days after eclosing, some young workers made erroneous decisions whereas after 3-4 days the decisions were always correct. To assess brain maturation, mitotic activity in the brains of workers was individually analyzed using BrdU injection immediately after the behavioral test. Neurogenesis appeared to be complete at eclosion, but in 3 of 8 young workers that showed incorrect behavioral decisions, mitotic activity was observed in the antennal lobe. In contrast, no mitotic activity was observed in the brains of young workers who correctly discriminated their foster queens from alien queens. These results suggest that in newly emerged ant workers the ability to discriminate queens coincides with the completion of cell proliferation in the brain. PMID- 12907878 TI - Mechanical injury and repair of cells. AB - OBJECTIVE: To concisely review the field of cell plasma membrane disruption (torn cell surface) and repair. MAIN POINTS: Plasma membrane disruption is a common form of cell injury under physiologic conditions, after trauma, in certain muscular dystrophies, and during certain forms of clinical intervention. Rapid repair of a disruption is essential to cell survival and involves a complex and active cell response that includes membrane fusion and cytoskeletal activation. Tissues, such as cardiac and skeletal muscle, adapt to a disruption injury by hypertrophying. Cells adapt by increasing the efficiency of their resealing response. CONCLUSION: Plasma membrane disruption is an important cellular event in both health and disease. The disruption repair mechanism is now well understood at the cellular level, but much remains to be learned at the molecular level. Cell and tissue level adaptational responses to the disruption either prevent its further occurrence or facilitate future repairs. Therapeutically useful drugs might result if, using this accumulating knowledge, chemical agents can be developed that can enhance repair or adaptive responses. PMID- 12907877 TI - Paradigm of the injury-repair continuum during critical illness. AB - OBJECTIVES: To define injury and repair in the context of critical illness. DESIGN: Review of the literature. RESULTS: The medical practice of critical care medicine has long been one of aggressive assistance and support: utilizing state of the art technology to support the functions of the human body until homeostasis is again achieved and the patient is functioning sufficiently to allow the support to discontinue. Injury is defined as the disruption of molecular, cellular, or organ functions resulting from an external or internal stimulus. Repair is defined as an adaptive process that occurs in response to injury and involves both local and systemic responses that serve to restore structure and regulation for the purpose of organ/tissue function. The study of injury in critical illness is now occurring "upstream," at the genetic and cellular levels, to understand how damaging effects of acute inflammation from injury can be prevented or modulated. CONCLUSION: To treat the patient with critical illness more effectively, it is important to understand both the cause of the insults and the repair processes triggered by the insults because both processes affect the eventual course of the critical illness. PMID- 12907879 TI - Vessel injury and capillary leak. AB - OBJECTIVE: To understand the mechanism of pathologic capillary leak in the critically ill patient. DESIGN: Review of normal and altered physiology of the microvasculature. Review of recent literature describing pathogenesis, mediators, and interventions influencing capillary leak and microvascular repair. SETTING: In vitro and in vivo studies, the latter including animal and human subjects. MEASUREMENTS AND MAIN RESULTS: Capillary leak with resultant edema develops in the critical care setting on the basis of perturbations in Starling's equation, primarily as a result of increased capillary permeability to larger molecules. This process is most likely fueled by inflammatory mediators or mechanical stress. Attempts to prevent or treat this process remain largely unsuccessful; resuscitation is more often symptomatic than therapeutic. Models of microvascular repair focus on discrete injury and may not be applicable to the recovery of capillary damage secondary to a systemic leak CONCLUSIONS: Our understanding of capillary leak syndrome remains fragmented and weighted toward specific mediators contributing to the leak. The implications of extensive edema and the mechanism by which it resolves continue to be the subject of speculation rather than study. PMID- 12907880 TI - Repair of metabolic processes. PMID- 12907881 TI - Molecular diagnostics of injury and repair responses in critical illness: what is the future of "monitoring" in the intensive care unit? AB - OBJECTIVE: To identify potential future means of monitoring injury and repair in critical illness. DESIGN: Review of the literature. RESULTS: Critically ill patients are monitored extensively and intensively by such means as hemodynamics, laboratory values, and radiologic studies. In general, however, the goal of monitoring has been to measure the degree of injury and to prevent further injury, rather than to measure repair. Measures of repair have been limited to phenotypic end points such as return of organ function, as measured by blood chemistry. In this article, I examine how it may be possible in the future to monitor the progress of repair using genomic and proteomic biomarkers. These types of monitors would enable clinicians to control the healing environment using real time, rapid biomarkers, and sophisticated techniques to target therapy to the patient's current inflammatory state, taking into account the genetic makeup of the patient and his or her likely response to a given drug. CONCLUSIONS: The rapidly evolving sciences of genomics, proteomics, computational biology, and complex system theory can be used a) to model critical illness; b) to model adaptive and maladaptive responses to critical illness; c) to tailor treatments to create an ideal inflammatory environment for repair and regeneration, taking into account the individual genetic contribution; and d) to monitor the progress of repair. The major obstacles to reaching these goals are technological, and experience suggests that they will be overcome. PMID- 12907882 TI - Injury and repair in lung and airways. AB - Acute lung injury (ALI) and acute respiratory distress syndrome (ARDS) are common causes of morbidity and mortality in the intensive care unit. ALI/ARDS occurs as a result of systemic inflammation, usually triggered by a microorganism. Activation of leukocytes and release of proinflammatory mediators from multiple cellular sources result in both local and distant tissue injury. Tumor necrosis factor-alpha and interleukin-1 beta are the best characterized of the proinflammatory cytokines contributing to ALI/ARDS and subsequent fibrosis. The ultimate clinical course of ALI/ARDS often is determined by the ability of the injured lung to repopulate the alveolar epithelium with functional cells. Death may occur when fibrosis predominates the healing response, as it results in worsening lung compliance and oxygenation. The rodent bleomycin model of lung fibrosis allows the use of molecular tools to dissect the cellular and subcellular processes leading to fibrosis. The elements of this response may provide therapeutic targets for the prevention of this devastating complication of ALI/ARDS. PMID- 12907883 TI - Mucosal repair in the gastrointestinal tract. AB - OBJECTIVE: The epithelial response to injury in the intestinal mucosa will be described. DESIGN: A comprehensive evaluation of the literature was performed to provide a thorough review of mucosal injury and repair. RESULTS: The intestinal mucosa is a rapidly proliferating sheet of epithelial cells that sustains injury in response to stresses ranging from physiologic daily digestive trauma to severe insults associated with ischemia, chemicals, and infection. Breaks in epithelial continuity impair mucosal barrier function, perturb normal absorptive and secretory transport properties, and render the host susceptible to local infection and distant organ pathology. Minor breaches are rapidly repaired by epithelial restitution, a process independent of cell proliferation. Restitution is regulated by a variety of cytokines and growth factors and is modulated by integrin-dependent interactions with the extracellular matrix. The intracellular mechanisms that control restitution are complex and involve signaling pathways that control dynamic remodeling of the actin cytoskeleton. Emerging understanding of reparative processes suggest several possible therapeutic strategies to enhance gastrointestinal wound healing. CONCLUSION: Minor epithelial injuries are repaired with the complex process of epithelial restitution independent of cell proliferation. PMID- 12907884 TI - Repair of full-thickness bowel injury. AB - OBJECTIVE: Colon surgery is more and more often performed in complex situations such as after trauma, under immunosuppression, or in the elderly. Even under optimal conditions, anastomosis fails in certain situations. The objective of this study was to demonstrate the normal phases of bowel healing and to review the local and systemic factors affecting healing with special attention to critical care variables such as major surgery, acute hemorrhage, and infections. DATA SOURCE: MEDLINE cited and/or published articles. DESIGN: Review analysis. RESULTS: Colon healing is a structured cascade of different phases that can be affected by a multitude of local (infection, ischemia) and systemic (diabetes, malnutrition, anemia, hypothermia, trauma) factors. The normal phases of repair, the resulting bursting pressure as an experimental index of healing, and the available published data on local and systemic factors affecting healing are summarized. CONCLUSION: Several local and systemic factors negatively affect bowel healing; there is still a small portion of patients who fail to heal, suggesting that intrinsic factors need to be analyzed. PMID- 12907885 TI - Healing responses of skin and muscle in critical illness. AB - OBJECTIVE: Many aspects of the care and underlying pathologies in patients suffering critical illness can detrimentally influence the normal healing processes of skin and soft tissues. Although a great diversity of pathologies exists, some aspects of the diseases and their treatments are common in critically ill patients. We aimed to identify some features, both common and specific, that could influence wound healing and the mechanisms by which they may do so. DESIGN: In this review, we first outline the biology of normal skin and muscle healing and then explore how critical illness may influence the normal healing cascade. FINDINGS: The healing of skin and skeletal muscle in critical illness is influenced by both underlying disease processes and the intensive care environment. Local and systemic factors can contribute to impaired healing, with the potential to prolong functional disability and increase the likelihood of wound complications. The frequency and number of soft tissue injuries derived from accidental injury, surgical intervention, and the need for invasive monitoring and therapies in the intensive care unit setting are likely to compromise the innate immunity and potentially further jeopardize the patient's ability to heal. Alterations in coagulation, tissue perfusion, inflammation, immune functioning, metabolism, nutrition, and drug therapies will influence healing responses by modifying the biological responses to tissue disruption. Locally, wound contamination, sepsis, tissue hypoxia, edema, and excessive or prolonged local pressure all have the potential to compromise soft tissue healing. One or more of these factors may be present at any time. CONCLUSION: The skin and soft tissues are vulnerable to both injury and compromised healing when a patient is critically ill and exposed to a critical care environment. The identification of risk factors may aid in forming and modifying treatment strategies when caring for the critically ill patient with soft tissue injuries. PMID- 12907887 TI - Molecular mechanisms of recovery from acute renal failure. AB - OBJECTIVES: Despite technological advances in renal replacement therapy over the past few years, acute renal failure in the intensive care unit remains associated with high morbidity and mortality rates. In this article I review recent research aimed at elucidating mechanisms of renal recovery from acute injury. DESIGN: Review of the literature. CONCLUSIONS: A number of peptide growth hormones are reviewed, including epidermal growth factor, insulin-like growth factor-1, thyroxine, hepatocyte growth factor, and bone morphogenetic protein-7 promote renal regeneration in model systems. Unfortunately, despite promising studies in animal models of toxin and ischemia-induced acute tubular necrosis, human studies have not shown any clinical benefit. However, several of these molecules have not been studied in clinical trials. Existing pharmacologic strategies have a limited role in renal recovery. Finally, several recent studies have focused on the effects of renal replacement therapy on renal recovery, but additional studies are needed to confirm and extend these results. PMID- 12907888 TI - Pancreas: healing response in critical illness. AB - OBJECTIVE: The pathophysiology of acute pancreatitis represents a diverse mix of congenital, hereditary, and acquired problems associated with or causing acute pancreatic inflammation. Acute pancreatitis is characterized by acinar cell injury that may involve regional and systemic inflammatory responses. The systemic manifestations of acute pancreatitis are responsible for the majority of pancreatitis-associated morbidity and are due to the actions of specific inflammatory cytokines. This report summarizes this pancreatic injury, the role of cytokines in the pathogenesis of acute pancreatitis, and the pancreatic healing response that follows. DESIGN: A comprehensive literature review of experimental pancreatitis as well as reports of cytokine involvement and healing response during clinical pancreatitis was performed. RESULTS: Histamine release, bradykinin generation, and cytokine release play a significant role during acute pancreatic inflammation. Following an experimental insult, there is rapid expression of tumor necrosis factor-alpha, interleukin-6, interleukin-1, and chemokines by pancreatic acinar cells and/or transmigrated leukocytes. Preventing the action of these mediators has a profound beneficial effect in experimental animals. Pancreatic fibrosis is a central histologic response after pancreatitis. Transient collagen deposition with acinar necrosis occurs in acute pancreatitis; in chronic pancreatitis, permanent and disorganized pancreatic fibrosis and parenchymal cell atrophy occur. CONCLUSIONS: Inflammatory mediators are responsible for the systemic manifestations of acute pancreatitis and the associated distant organ dysfunction. After the acute injury, regeneration or pancreatic repair is characterized by decreased release of proinflammatory mediators and decreased infiltrating inflammatory cells. Differentiation and proliferation of pancreatic myofibroblasts or "stellate" cells may be responsible for increased extracellular matrix production. The predictable nature in which the inflammation and fibrosis are produced may stimulate novel approaches to disease treatment. PMID- 12907889 TI - Practice parameters for the treatment of patients with dominantly inherited colorectal cancer (familial adenomatous polyposis and hereditary nonpolyposis colorectal cancer). PMID- 12907886 TI - Mechanisms of immune resolution. AB - Initially after injury, the innate/proinflammatory and some aspects of the acquired immune response are up-regulated to maintain a defense against foreign pathogens, clear tissue debris present at the wound site, and orchestrate aspects of tissue remodeling, cell proliferation and angiogenic process, associated with the wound response. However, for proper wound healing to progress, this initial inflammatory response has to be regulated or shut down so as to allow for the reestablishment of matrix, recellularization, and tissue remodeling. Inability to properly resolve the extent of innate/acquired response at a site of injury can lead to poor wound healing, immune suppression, and recurrent infectious episodes. This review attempts to summarize information on regulatory mechanisms that are thought to be involved in controlling/resolving innate or acquired immune responses so as to provide a framework for use in thinking about the impact these processes and their manipulation may have on wound healing and its potential management. PMID- 12907890 TI - Clinical value of preoperative mechanical bowel cleansing in elective colorectal surgery: a systematic review. AB - PURPOSE: This study was designed to establish scientific evidence for and clinical results of preoperative mechanical bowel cleansing before elective colorectal surgery. METHODS: Systematic literature searches in electronic databases, conference proceedings, and hand searches of reference lists of previously retrieved literature without any language restrictions were used. Only randomized trials were included. A quality assessment of each retrieved trial was performed. Outcome measures were surgical infections, mortality, and anastomotic dehiscence. Meta-analyses of the selected trials were performed using the Peto odds ratio. RESULTS: The results of each outcome were as follows. 1). Overall anastomotic leakage-six studies: 5.5 percent with cleansing compared with 2.9 percent without cleansing; odds ratio 1.94, 95 percent confidence interval: 1.09 to 3.43 (P = 0.02). 2). Peritonitis-three studies: 5.1 percent with cleansing compared with 2.8 percent without cleansing; odds ratio 1.90, 95 percent confidence interval: 0.78 to 4.64 (not significant). 3). Wound infection-six studies: 7.4 percent with cleansing compared with 5.7 percent without cleansing; odds ratio 1.34, 95 percent confidence interval: 0.85 to 2.13 (not significant). CONCLUSIONS: There is no evidence in the literature for beneficial effects from the use of bowel cleansing before elective colorectal surgery. Cleansing seems to be associated with an increased risk of more anastomotic dehiscence. Further studies stratifying between rectal and colonic surgery are warranted. PMID- 12907891 TI - Met-analyses. PMID- 12907892 TI - Lack of influence of cytokeratin-positive mini micrometastases in "Negative Node" patients with colorectal cancer: findings from the national surgical adjuvant breast and bowel projects protocols R-01 and C-01. AB - PURPOSE: Results of the few extant reports concerning the clinical significance of so-called "occult micrometastases" of lymph nodes of patients with Dukes A and B colorectal cancer have been variable. We examined the presumably negative nodes of a larger cohort of such patients who were enrolled in the National Surgical Adjuvant Breast and Bowel Project clinical trials R-01 and C-01 for the influence of what we preferably designate as nodal mini micrometastases on parameters of survival. METHODS: Mini micrometastases were detected by immunohistochemical staining of the original lymph node sections with anticytokeratin A1/A3 in a total of 241 Dukes A and B patients with rectal and 158 with colonic cancers. Their frequency, as well as that of nuclear and histologic grades, and an estimation of their relationship to relative risks were correlated with overall and recurrence-free survival by univariate and multivariate analyses. RESULTS: Nodal mini micrometastases were detected in 73 of 399 (18.3 percent) patients of this cohort. They failed to exhibit any significant relationship to overall or recurrence-free survival. No association between the assessments of tumor differentiation and mini micrometastases was found. Nuclear and histologic grades also failed to further discriminate overall or recurrence-free survival in patients with A or B stages of colonic or rectal cancers in this cohort. CONCLUSION: The immunohistochemical demonstration of nodal mini micrometastases failed to discriminate high- and low-risk groups of patients with colorectal cancer who were designated as being node-negative after routine pathologic examination. PMID- 12907894 TI - Sentinel node biopsy for squamous-cell carcinoma of the anus and anal margin. AB - PURPOSE: The majority of anal tumors are squamous-cell carcinomas. These may be tumors of the anal canal or margin. They are best treated by combination of chemotherapy and radiotherapy. T1 and T2 tumors in this regime do not receive radiotherapy to the inguinal regions despite approximately 5 to 10 percent incidence of inguinal lymph node involvement. If the nodal status of the inguinal region could be accurately assessed, then a more tailored radiotherapy regime may be given. This article describes a novel method of assessment of the status of the inguinal lymph nodes in patients. METHODS: Patients with anal squamous-cell carcinoma had four injections of 0.2 ml of antimony sulfide (30 MBq) around the tumor. Under a gamma camera, a distant high-intensity signal was located, and this point was marked on the overlying skin using an indelible ink pen. In the operating theater, patent blue dye was injected all around the tumor. The localized lymph node was removed and sent for histopathology. RESULTS: This procedure was performed on 12 patients. The sentinel node was localized to the inguinal region and removed in eight of these patients. In two patients, metastatic squamous-cell carcinoma was identified histologically in the sentinel node. CONCLUSION: We advocate that this as a safe technique for detecting metastatic disease in the inguinal nodes in patients with anal squamous-cell carcinoma. PMID- 12907896 TI - Sentinel lymph node procedure in patients with epidermoid carcinoma of the anal canal: early experience. AB - PURPOSE: This study was conducted to assess the feasibility of the sentinel lymph node procedure in patients with epidermoid carcinoma of the anal canal. METHODS: Between February 2001 and November 2002, 14 patients with epidermoid carcinoma of the anal canal and no clinical evidence of inguinal involvement were prospectively enrolled in the study. The sentinel lymph node procedure consisted of a combination of preoperative lymphoscintigraphy with technetium 99m dextran 500 injected around the tumor and intraoperative detection of the sentinel node with a gamma probe. Patent blue V dye was also injected at the periphery of the tumor to facilitate direct identification of the blue-stained lymph node. After removal, the sentinel node was studied by hematoxylin and eosin staining and immunohistochemistry for pancytokeratins (antigen A1 and A3). RESULTS: Detection and removal of sentinel lymph nodes was possible in all patients. There was no correlation between tumor size and pattern of lymphatic drainage to the groin. Tumors located in the midline of the anal canal gave rise to bilateral sentinel nodes in eight of nine cases. In total, 23 sentinel lymph nodes were removed. One patient (7.1 percent) had a node identified as positive for metastatic carcinoma on immunohistochemical staining. Surgical complications were minimal. CONCLUSIONS: The standardized technique was safe and highly effective in sampling inguinal sentinel lymph nodes in carcinoma of the anal canal. It also proved to be useful as an instrument to detect micrometastatic deposits in clinically normal nodes. Our early results suggest the sentinel lymph node procedure may have a role in guiding a more selective approach for patients with anal cancer. Additional studies in a larger patient population to determine the sensitivity and specificity of this method are warranted. PMID- 12907897 TI - Doctor-patient communication: results of a four-year prospective study in rectal cancer patients. AB - PURPOSE: The purpose of the study was to examine the effect of communication on rectal cancer patients' quality of life over four years. Previous studies have either used short follow-up periods or examined only certain aspects of quality of life, such as anxiety and depression. METHODS: In a prospective, observational study, rectal cancer patients, recruited by clinicians over a two-year period, were sent questionnaires over four years. The clinical details of these patients were recorded by the Munich Cancer Registry. The psychological scores from the European Organization for Research and Treatment of Cancer QLQ-C30 and CR38 were the main outcome variables. RESULTS: Thirty-nine percent of the sample reported that some aspect of the communication they received was unclear (incomprehensible or too little). More than 60 percent wished to speak more with their physician. Younger patients and those in larger hospitals were more likely to report unclear communication (P < 0.05). Analyses of covariance, controlled for age, gender, adjuvant therapy, stoma, education, clinic, and comorbidity, demonstrated that role, emotional, and social functioning scores were consistently lower in patients reporting unclear communication. Additionally, these patients experienced more problems sleeping, poorer body image, more financial worries, and a worse future perspective. Repeated measures analyses indicated that sleeping problems and emotional and social functioning difficulties persisted for at least three years. CONCLUSION: Reports of unclear communication were associated with poorer quality of life in rectal cancer patients without disease progression. PMID- 12907898 TI - Outcome of laparoscopic surgery for rectal cancer in 101 patients. AB - PURPOSE: This study was conducted to investigate the feasibility of laparoscopic resection of rectal cancer and to compare early outcome data with the results of the conventional technique. METHODS: From January 1996 to March 2002, 435 patients with primary rectal cancer were operated on at our institution. Low risk, small rectal tumors treatable by local excision, rectal cancer recurrences, and emergency cases were excluded from the analysis. Three hundred thirty-four patients were operated on by the conventional open approach. One hundred one selected patients underwent surgery by the laparoscopic technique. RESULTS: Because of the selection process, significantly more patients with early tumor stages were operated on by laparoscopy. There were no differences in mean operation time, morbidity, mortality, or the anastomotic leakage rate; however, the need for intraoperative transfusion, mean stay in the intensive care unit, and length of hospital stay were reduced significantly. CONCLUSIONS: In terms of the intraoperative and early postoperative course, the laparoscopic resection of rectal cancer in a selected cohort of patients compares favorably with the open technique. Because follow-up time is limited to date, only very preliminary information can be given on tumor-related outcome data. However, these preliminary data appear to suggest that rectal cancer resection can be performed by laparoscopy in accordance with established principles of cancer therapy and that port-site metastases are not a relevant clinical problem. Prospective, randomized trials are required to determine whether the laparoscopic approach will play a significant role in the treatment of rectal cancer in the future. PMID- 12907899 TI - Tumor budding at the invasive margin can predict patients at high risk of recurrence after curative surgery for stage II, T3 colon cancer. AB - PURPOSE: The aim of this study was to identify indicators that can predict patients at high risk of tumor recurrence in Stage II, T3 colon cancer. METHODS: A total of 138 patients classified as Stage II, T3 underwent curative resection of colon cancer between 1981 and 1993. Clinical variables included age, gender, bowel obstruction, tumor location, and emergency presentation. For each colon tumor specimen, the following histopathological variables were assessed: maximum tumor diameter (<5 vs. >or=5 cm), depth, tumor grade (well and moderate vs. other), lymphatic and venous invasion (absent vs. present), perineural invasion, tumor necrosis, and tumor margin (expanding vs. infiltrating). We also categorized tumor budding, defined as a single cancer cell or small clusters of undifferentiated cancer cells in the invasive frontal lesion, into two categories: none or minimal (BD-1), and moderate or severe (BD-2). Univariate analysis for factors regarding recurrence and disease-specific survival were performed with the logistic regression model and the log-rank test. RESULTS: Among the factors analyzed, tumor budding was the only factor that was significantly associated with recurrence and survival. The numbers of patients with BD-1 and BD-2 tumors were 111 and 27, respectively. Forty-eight percent of BD-2 tumor patients developed recurrence, compared with 4.5 percent of BD-1 tumor patients (P < 0.0001). The cumulative disease-specific survival rates at five years for patients with BD-1 and BD-2 tumors were 98 and 74 percent, respectively (P < 0.0001). CONCLUSION: The presence of moderate or severe budding at the invasive margin in Stage II, T3 colon cancer indicated a high risk of tumor recurrence after curative surgery, providing useful information for the decision regarding postoperative adjuvant chemotherapy. PMID- 12907900 TI - Adjuvant therapy with protein-bound polysaccharide K and tegafur uracil in patients with stage II or III colorectal cancer: randomized, controlled trial. AB - PURPOSE: Intravenous fluorouracil and leucovorin for six to eight months is currently a standard adjuvant treatment for Stage III colon cancer; however, this regimen is complex, inconvenient, and has a high intolerability. Adjuvant chemotherapies are claimed for objective response rates with an acceptable safety profile and complexity. We investigated the benefits of oral protein-bound polysaccharide K added to oral tegafur/uracil on curatively resected Stage II or III colorectal cancer. METHODS: We prospectively randomized 207 patients to treatments of either oral 3.0 g protein-bound polysaccharide K plus 300 mg tegafur/uracil or 300 mg tegafur/uracil alone for two years following 12 mg/m2 and 8 mg/m2 mitomycin treatment on postoperative Days 1 and 2, respectively. The primary end points were disease-free and overall survival, and recurrence rates. RESULTS: Three (1.4 percent) patients were declared ineligible, and three patients did not start treatment. In total, 201 patients were analyzed. The three year, disease-free survival rate was 80.6 percent (standard error = 3.4 percent) in the protein-bound polysaccharide K group (P = 0.02) compared with 68.7 percent (SE = 5.7 percent) in the control group after a median follow-up of 3.7 years. The estimated relative risk of recurrence in the control group was 1.87 (95 percent confidence interval, 1.10-3.20) at three years. The three-year, overall survival rate was 87.3 percent (standard error = 2.9 percent) in the protein bound polysaccharide K group and 80.6 percent (standard error = 4.8 percent) in the control group (P = 0.24). The three-year, overall survival rate in 80 pathological TNM Stage III patients was 83.0 percent (standard error = 5.2 percent) in the protein-bound polysaccharide K group and 59.3 percent (standard error = 9.5 percent) in the control group (P = 0.02). Protein-bound polysaccharide K prevented distant metastases (P = 0.05), particularly lung metastases (P = 0.01). The incidence of adverse effects was minimal, and compliance was good. CONCLUSION: Adjuvant therapy using a combination of oral protein-bound polysaccharide K and tegafur/uracil is highly effective in preventing the recurrence of colorectal cancer in Stage II or III patients, and increases overall survival in pathological TNM Stage III. These results will be a sufficient proof to conduct a larger study to compare tegafur/uracil/protein bound polysaccharide K with 5-fluorouracil/leucovorin. PMID- 12907901 TI - Microsatellite instability and mutations in DNA mismatch repair genes in sporadic colorectal cancers. AB - PURPOSE: This study was designed to investigate the frequency of mutations in DNA mismatch repair genes in sporadic colorectal cancers. METHODS: Genomic DNAs procured from paraffin blocks of the pathologic specimens from 230 consecutive patients with colorectal cancer were examined for their microsatellite instability status using a mononucleotide microsatellite marker, BAT-26, and also evaluated expressions of hMLH1, hMSH2, and hMSH6 proteins by immunohistochemical staining. Any of these 230 patients did not have family histories of hereditary nonpolyposis colorectal cancer, familial adenomatous polyposis, colorectal cancer, or hereditary nonpolyposis colorectal-related cancers, such as endometrial, small bowel, and ureteral and renal pelvic cancers. When microsatellite instability was positive, mutations in the simple repeated sequences of TGF-betaRII, BAX, IGF IIR, hMSH3, and hMSH6 genes were examined. In microsatellite instability-positive or staining-negative cases, polymerase chain reaction-single-strand conformation polymorphism and DNA sequencing detected mutations of hMLH1, hMSH2, and hMSH6 genes. If mutations were found in tumor tissue samples, we tested for a germline mutation with a microdissected corresponding normal tissue. RESULTS: Among 230 cases of sporadic colorectal cancer, 21 (9.1 percent) manifested microsatellite instability. In the immunohistochemical staining, 20 (8.6 percent) showed loss of expressions. All 20 staining-negative cases were microsatellite instability-positive. Only 1 of 21 (4.8 percent) microsatellite instability-positive cases showed intact staining for three proteins. The frame-shift mutations of the simple repetitive sequences were found in 17 cases (81.0 percent) in TGF-betaRII, 11 (52.4 percent) in BAX, 5 (23.8 percent) in IGF IIR, 7 (33.3 percent) in hMSH3, and 8 (38.1 percent) in hMSH6 genes. Germline mutation was observed in only one case, which accounts for 4.8 percent among positive microsatellite instability and 0.4 percent of total patients, and was found in hMSH2. Five somatic mutations (2 in hMLH1, 2 in hMSH2, and 1 in hMSH6) also were found. CONCLUSION: The results indicated that a germline mutation of DNA mismatch repair gene was a rare event in sporadic colorectal cancers. PMID- 12907902 TI - Comparison of ultrasound and manometric sphincter length and incontinence scores. AB - PURPOSE: Fecal incontinence can be assessed by a wide variety of means that vary in their clinical relevance. This study assesses the correlation between two methods of recording sphincter length and incontinence scores. METHODS: A retrospective review of results from 1999 to 2001 of 137 patients with fecal incontinence was undertaken on patients who had both ultrasound and manometric assessment. Incontinence scores, ultrasonic sphincter length, and manometrically derived sphincter length were recorded. The data were analyzed using the Spearman rank correlation coefficient (r(s)) for nonparametric data. RESULTS: There was negative correlation between incontinence scores and manometric sphincter length (r(s) = -0.27; P = 0.002) but not with ultrasonic length (r(s) = -0.08; P = 0.38). There was correlation between manometric and ultrasonic lengths (r(s) = 0.21; P = 0.02). In males, all three correlated but none of the values were significant (n = 12). Internal and external sphincter defects had no effect on correlation coefficients. CONCLUSION: Given that all measurements include a degree of subjectivity, manometry more closely correlates with the fecal incontinence scores than ultrasound measurements. There seems to be little benefit in measuring the sphincter length with ultrasound. Manometric sphincter length measurement seems to have a role in assessing patients with fecal incontinence. PMID- 12907903 TI - Anal sphincter injury, fecal and urinary incontinence: a 34-year follow-up after forceps delivery. AB - PURPOSE: This study was designed to determine the long-term outcome of forceps delivery in terms of evidence of anal sphincter injury and the incidence of fecal and urinary incontinence. METHODS: Women who delivered in 1964 were evaluated by using endoanal ultrasound, manometry, and a continence questionnaire. Women delivered by forceps were matched with the next normal delivery and elective cesarean delivery in the birth register. RESULTS: The women's overall obstetric history was evaluated. Women who had ever had a forceps delivery (n = 42) had a significantly higher incidence of sphincter rupture compared with women who had only unassisted vaginal deliveries (n = 41) and elective cesarean sections (n = 6) (44 vs. 22 vs. 0 percent; chi-squared 7.09; P = 0.03). There was no significant difference in the incidence of significant fecal incontinence between the three groups (14 vs. 10 vs. 0 percent) or significant urinary incontinence (7 vs. 19 vs. 0 percent). CONCLUSION: Anal sphincter injury was associated with forceps delivery in the past; however, significant fecal and urinary incontinence was not. PMID- 12907904 TI - Functional results after the Orr-Loygue transabdominal rectopexy for complete rectal prolapse. AB - PURPOSE: The aim of this study was to assess functional results after Orr-Loygue transabdominal rectopexy for complete rectal prolapse. METHODS: Thirty-one consecutive patients operated on for complete rectal prolapse between 1995 and 1998 were evaluated preoperatively and postoperatively with regard to anal incontinence, constipation, evacuation difficulties, and overall satisfaction with a standardized questionnaire. Anal continence assessment was based on a clinical scoring system. RESULTS: After a mean follow-up of 28 +/- 13 (range, 12 57) months, no prolapse recurred. Preoperative and postoperative rates of incontinence were 81 percent (25/31) and 55 percent (17/31), respectively (P < 0.03). Continence improved in 24 (96 percent) of the 25 patients who were incontinent before surgery. The mean incontinence score decreased from 11.7 +/- 7.8 preoperatively to 3.2 +/- 4.2 postoperatively (P < 0.001). The self-reported constipation rate was 61 percent (19/31) before surgery and 71 percent (22/31) after surgery (P = nonsignificant). Constipation appeared or worsened in 16 patients (52 percent), whereas it disappeared or improved in 8 (26 percent; P = nonsignificant). Evacuation difficulties increased significantly after surgery from 23 percent (7/31) to 61 percent (19/31; P < 0.003). Ninety-seven percent of patients reported good or very good satisfaction. CONCLUSIONS: Transabdominal Orr Loygue rectopexy resulted in improved continence and a high satisfaction level. Despite a significant postoperative increase in evacuation difficulties, only a 10 percent (nonsignificant) increase in the self-reported constipation rate was observed. PMID- 12907905 TI - Botulinum toxin (botox) reduces pain after hemorrhoidectomy: results of a double blind, randomized study. AB - PURPOSE: Pain after hemorrhoidectomy appears to be multifactorial and dependent on individual pain tolerance, mode of anesthesia, postoperative analgesia, and surgical technique. Spasm of the internal sphincter is believed to play an important role. The aim of this study was to assess the role of botulinum toxin in reducing pain after Milligan-Morgan hemorrhoidectomy. METHODS: This was a double-blind study of 50 consecutive patients undergoing Milligan-Morgan hemorrhoidectomy and assigned to an internal sphincter injection of 0.4 ml of solution containing either botulinum toxin (20 U; Botox) or normal saline. Patients were managed according to standardized perioperative analgesic and laxative regimens. Pain was assessed by use of daily visual analog scores and analgesia requirements for the first seven postoperative days. RESULTS: Patients randomized to receive botulinum toxin had lower daily average and maximal visual analog scores throughout the study period. The difference reached significance on both Day 6 (P < 0.05) and Day 7 (P < 0.05). There was no significant difference (P = 0.12) in morphine requirements in the first 24 hours (botulinum group, 16 (range, 6-27) mg; placebo arm, 22 (range, 13-41) mg). Patients who received Botox used 19 (range, 8-36) coproxamol tablets in the first seven days after surgery compared with 23 (range, 10-40) in the placebo arm (P = 0.63). CONCLUSIONS: Those patients who had botulinum toxin had significantly less pain toward the end of the first week after surgery. Reduction in spasm within the internal sphincter is the presumed mechanism of action. This is the first reported randomized, controlled trial using botulinum toxin in hemorrhoidectomy. PMID- 12907906 TI - Topical cidofovir (HPMPC) is an effective adjuvant to surgical treatment of anogenital condylomata acuminata. AB - PURPOSE: Human papilloma virus infections of the anogenital region are very common and cause condylomata acuminata; cervical, penile, vulvar, or perianal intraepithelial neoplasia; and more rarely, invasive cancer. The currently available therapies often result in painful, extensive, slow-healing ulcerations and frequent early relapses. This study was aimed at determining the efficacy of topical application of the antiviral agent cidofovir at 1 percent. METHODS: Twenty patients treated with coagulations were compared with 27 patients treated with cidofovir. Lesions refractory to cidofovir were cleared up with additional coagulations. The number of patients previously treated for condylomata did not differ between the two groups. Significantly more patients treated with cidofovir, however, had an impaired immune status (37 percent) compared with the patients treated with coagulations (5 percent). RESULTS: Cidofovir alone cured the lesions in 32 percent of the patients and induced partial regression in 60 percent. However, in smokers, complete resolution of the condylomata occurred only in 16.6 percent compared with 66 percent of nonsmokers (P = 0.03). The number of coagulation sessions was much lower (P < 0.0005) in the cidofovir treated group (1 +/- 0.8 vs. 2.9 +/- 2). Furthermore, the relapse rate was significantly lower in the cidofovir group (3.7 vs. 55). All recurrences in the electrocoagulation group occurred within four months of confirmed lesion clearance. Topical applications of cidofovir 1 percent were well tolerated. Thirty-three percent of the patients reported only mild pain caused by erosive dermatitis. In contrast, coagulations caused painful ulcerations that necessitated the use of analgesics in all patients treated this way. CONCLUSIONS: Topical applications of cidofovir, an antiviral compound with activity against human papilloma virus, is effective in the majority of patients with perianal condylomata and is a valuable adjuvant to surgical treatment of these lesions. PMID- 12907908 TI - Population-based incidence of complicated diverticular disease of the sigmoid colon based on gender and age. AB - PURPOSE: The purpose of this study was to characterize the gender and age differences in patients with clinically symptomatic sigmoid diverticular disease requiring surgery. METHODS: All surgical patients hospitalized with proven diverticular disease requiring sigmoid resection from January 1988 to January 1998 were reviewed. RESULTS: A total of 934 patients requiring surgical resection for diverticular disease were admitted. There were 443 men and 491 women with an average age of 64. Forty-nine patients presented with massive rectal bleeding (males, 3.6 percent; females, 1.6 percent), 329 with chronic diverticulitis (males, 15.8 percent; females, 19.3 percent), 61 with obstructive symptoms (males, 2.7 percent; females, 3.9 percent), 148 with fistulas (males, 8.0 percent; females, 7.8 percent), 170 with perforation (male, 8.7 percent; female, 9.4 percent), 79 with abscess (males, 4.0 percent; females, 4.5 percent), 59 with stricture (males, 2.2 percent; females, 4.0 percent), and 39 with acute diverticulitis (males, 2.2 percent; females, 1.9 percent). Overall, patients younger than 50 presented more often with chronic or recurrent diverticulitis. CONCLUSIONS: Female patients present, on average, five years later than male with complications requiring surgery. Overall, men have a higher incidence of bleeding (P = 0.015), whereas women present more often with stricture and obstruction (P = 0.02). Young males present more with fistula (P = 0.03), whereas older males present with bleeding (P = 0.001). Young females present with perforation (P = 0.002), and older females present with chronic diverticulitis (P = 0.04) and stricture (P = 0.04). PMID- 12907909 TI - Muscle fragment welding for control of massive presacral bleeding during rectal mobilization: a review of eight cases. AB - INTRODUCTION: Although massive presacral bleeding during rectal mobilization is uncommon, it can rapidly destabilize a patient. Traditional attempts at control include tamponade with pelvic packing and application of sacral thumbtacks. The aim of this review is to describe the anatomic basis of injury and summarize our experience with this challenging problem, with emphasis on the simple, readily available, effective technique of rectus abdominis muscle fragment welding. METHODS: A retrospective review of eight patients who underwent muscle fragment welding for presacral bleeding incurred during rectal mobilization was undertaken. This technique involves harvesting a small piece of rectus abdominis muscle, which is held in place with a forceps to occlude the bleeding site. Electrocautery adjusted to the highest setting is then applied to the forceps to "weld" closed the bleeding point. RESULTS: Control of presacral bleeding was achieved in all eight patients (3 males) with this technique without complications attributable to this method. Previous attempts at pelvic packing failed in all eight patients. CONCLUSION: Muscle fragment welding is a safe, readily available, and highly effective method of controlling massive presacral bleeding. PMID- 12907910 TI - Safety and outcome of use of nonabsorbable mesh for repair of fascial defects in the presence of open bowel. AB - PURPOSE: Historically, there has been reluctance to use nonabsorbable synthetic mesh for repair of abdominal-wall defects in an operative field in which the presence of open bowel is accompanied by the potential for contamination. Some believe the risk of wound infection and mesh removal in this setting to be unacceptably high. The purpose of this study was to evaluate the safety and efficacy of nonabsorbable mesh used for hernia repair in the presence of a stoma or at the time of colon resection. METHODS: All patients undergoing elective surgical implantation of mesh with concomitant open bowel from 1987 to 2001 were retrospectively reviewed. Computer database identified all patients undergoing parastomal hernia repair, ventral hernia repair with a stoma present, hernia repair with concomitant bowel resection, and colostomy closure with repair of hernia. No patients so identified were excluded. Follow-up was attained on all patients by chart review and telephone survey. The data was statistically analyzed by chi-squared test using a P value of <0.05 for statistical significance. RESULTS: Twenty-nine patients were identified as having undergone 30 elective hernia repairs using nonabsorbable mesh. The repairs were performed in the presence of a stoma or in conjunction with bowel resection. All patients received bowel preparation. Included were 11 patients undergoing parastomal hernia repair (37 percent), 14 patients undergoing ventral hernia repair in the setting of open bowel (47 percent), and 5 patients in whom mesh repair of ventral and parastomal hernias were performed simultaneously (16 percent). Hernias recurred in 13 patients (43 percent). Overall recurrence for mesh repair at a parastomal site was 63 percent; overall recurrence at an incisional hernia site was 21 percent. The risk of wound complications after mesh placement in the setting of open bowel was assessed. Wound seromas developed after surgery in four patients (13 percent). Seromas were all treated successfully by aspiration. Wound infections occurred after surgery in two patients (7 percent). Wound infection occurred exclusively in sites of parastomal repair representing 2 of 16 (13 percent) of parastomal hernia sites. Infection with fistula necessitated mesh removal in one of these two cases. No chronic sinuses were observed. Incidences of recurrence and wound infection were statistically independent of type of hernia, variety of mesh, or operative approach. CONCLUSION: After bowel preparation, nonabsorbable mesh can be used for elective repair of incisional hernia in the presence of open bowel with an expectation of minor morbidity, minimal risk of infection, and an acceptable rate of recurrence. Nonabsorbable mesh can be used for elective repair of parastomal hernia in a similar setting with a low risk of infection independent of surgical approach. Although safe, local mesh repair of parastomal hernia was, in this study, accompanied by a high rate of recurrence. PMID- 12907911 TI - Outcome of patients undergoing liver transplantation for primary sclerosing cholangitis. AB - PURPOSE: The purpose of this study was to determine the outcome of patients with inflammatory bowel disease who underwent liver transplantation for primary sclerosing cholangitis. METHODS: All patients who underwent liver transplantation for primary sclerosing cholangitis at our institution were identified. A review of patients' hospital and office charts was performed; all patients were then contacted, and a detailed survey was administered by telephone. RESULTS: Sixty nine patients were identified. There were 53 males (76.8 percent) and 16 females, with a mean age of 45.3 (+/- 13.3) years. Fifty-two (75.4 percent) of the 69 patients had documented inflammatory bowel disease; of these, 40 had ulcerative colitis (76.9 percent), 11 had Crohn's disease, and 1 had indeterminate colitis. Thirty-one patients (60 percent) were diagnosed with inflammatory bowel disease before primary sclerosing cholangitis, with a mean interval to diagnosis of primary sclerosing cholangitis of 10.8 (+/- 10.3) years. Seven patients had both diagnoses made at roughly the same time, and 14 patients initially were diagnosed with primary sclerosing cholangitis and subsequently were found to have inflammatory bowel disease, with a mean interval of 5.2 (+/- 4.4) years; 5 (35.7 percent) of those 14 patients were only diagnosed with inflammatory bowel disease after their liver transplant. The mean time from diagnosis of primary sclerosing cholangitis to liver transplantation was 6.1 (+/- 4.9) years. Since their transplant, 30.8 percent of patients rated their colitis as worse, 38.5 percent felt it was unchanged, and 30.8 percent felt that their colitis was better controlled. Eight (15.4 percent) of the 52 patients with inflammatory bowel disease denied having any knowledge of an increased risk of colorectal neoplasia. Four patients have required colectomy for colorectal neoplasia after liver transplantation, at a mean of 4.7 years after transplantation. Of the patients with inflammatory bowel disease, 42 (80.1 percent) had at least 1 posttransplant surveillance colonoscopy. Eight of the remaining ten patients had a colectomy, leaving only two patients (3.8 percent) who had not been surveyed. However, only 32 (61.5 percent) of the patients with inflammatory bowel disease have been on a surveillance regimen that would approximately conform to current screening recommendations. CONCLUSIONS: The activity of inflammatory bowel disease after transplantation is highly variable. Patients appeared to lack knowledge of their increased risk for colorectal neoplasia. Colorectal cancer is an uncommon but important complication in patients after liver transplantation for primary sclerosing cholangitis, and ongoing surveillance is required. Patients may require education to increase their awareness of the cancer risk and compliance with surveillance. PMID- 12907912 TI - Comparison of conventional and laparoscopic ileocolic resection for Crohn's disease. AB - PURPOSE: The purpose of this study was to compare open with laparoscopic ileocolic resection in selected patients with refractory Crohn's disease confined to terminal ileum and cecum in terms of small-bowel obstruction and recurrence rates at a follow-up of five years. METHODS: Thirty-nine patients who underwent laparoscopic ileocolic resection during a four-year period were compared with 53 patients who had previously undergone open ileocolic resection by the same surgeons at the same institution. Small-bowel obstruction was any clinical evidence of obstruction requiring hospital admission, regardless of recurrent disease or the need for surgery. Recurrence was defined as histologically proven Crohn's disease requiring reoperation and assessed by the actuarial method. Data were presented as medians and ranges and analyzed with Fisher's exact test, Student's t-test, Wilcoxon's rank-sum test, and log-rank test. RESULTS: Open ileocolic resection and laparoscopic ileocolic resection patients were well matched for age, gender, body mass index, American Society of Anesthesiologists grade, smoking, length of time from diagnosis to index operation, site of disease, preoperative medical treatment, previous abdominal surgery, indications for surgery, anastomosis configuration, microscopical involvement of resection margins, and postoperative complications (9.4 vs. 10.2 percent). Operating time was longer for laparoscopic ileocolic resection patients (105 (60-120) vs. 185 (130-210) min, P < 0.001). Hospital stay was shorter for laparoscopic ileocolic resection patients (11.2 (5-20) vs. 5.6 (3-11) days, P < 0.001). At five-year follow-up, 90.5 percent of open ileocolic resection patients and 92.3 percent of laparoscopic ileocolic resection patients were available. Five-year small-bowel obstruction rates were 35.4 and 11.1 percent, respectively, in open ileocolic resection and laparoscopic ileocolic resection patients (17 vs. 4 patients, P = 0.02). Five-year-recurrence rates were 29.1 (14/48) and 27.7 percent (10/36). Median time to recurrence was 48 and 56 months, respectively, following open ileocolic resection and laparoscopic ileocolic resection (P = 0.9104). CONCLUSION: When compared with open ileocolic resection, laparoscopic ileocolic resection led to lower five-year small-bowel obstruction rates in selected patients with ileocecal Crohn's disease. Five-year recurrence rates did not differ. PMID- 12907913 TI - Adenocarcinoma in the anal transitional zone after ileal pouch for ulcerative colitis: report of a case. AB - This article reports the seventh known case of adenocarcinoma arising in or adjacent to an ileal pouch after proctocolectomy for ulcerative colitis. It is the second reported case of adenocarcinoma in the anal transitional zone in this setting. A literature review is presented of the six previous cases published, and on the wider subject of how to best manage the anal transitional zone. It is concluded that this is a rare, but catastrophic, event with a potentially poor prognosis and can occur late (more than a decade) after the original surgery. All ileal pouches performed for ulcerative colitis should be followed for extended periods. The development of dysplasia necessitates close follow-up, including regular biopsies, and local excision and pouch advancement can manage persistent dysplasia. When operating for dysplasia or cancer, biopsies of the anal transitional zone should be performed or consideration given to mandatory mucosectomy. PMID- 12907914 TI - Vertebral venous air embolism: an unusual complication following colonoscopy: report of a case. AB - Although fiberoptic colonoscopy has gained wide popularity as a diagnostic and therapeutic tool, there remains an inherent complication rate following colonoscopic evaluation. Endoscopically induced bowel perforation and uncontrolled bleeding often necessitate immediate surgical intervention. Another often-unrecognized complication is the introduction of air into the vertebral venous system. A case of vertebral venous air embolism after routine diagnostic colonoscopy is reported with a review of current literature. PMID- 12907915 TI - Conundrum of the cocoon: report of a case and review of the literature. AB - Idiopathic, sclerosing, encapsulating peritonitis, or abdominal cocoon, is a rare cause of bowel obstruction in the elderly. A 65-year-old male patient presented with acute bowel obstruction caused by torsion of the encapsulated small bowel. He was treated successfully with subtotal excision of the fibrocollagenous membrane and small bowel resection. Despite anecdotal reports of a preoperative diagnosis, in the majority of cases, sclerosing, encapsulating peritonitis is a fortuitous finding. A better awareness of this condition may facilitate preoperative diagnosis. PMID- 12907916 TI - Botulinum toxin in the treatment of chronic anal fissure. PMID- 12907917 TI - Fibrin glue and anal fistulas. PMID- 12907918 TI - What mice can do for us: extending the field of neuroscience. PMID- 12907919 TI - Sharing genes? How mice can help us understand our biology. PMID- 12907920 TI - New lessons from knockout mice: The role of serotonin during development and its possible contribution to the origins of neuropsychiatric disorders. AB - Serotonin (5-HT) modulates numerous processes in the central nervous system that are relevant to neuropsychiatric function and dysfunction. It exerts significant effects on anxiety, mood, impulsivity, sleep, ingestive behavior, reward systems, and psychosis. Serotonergic dysfunction has been implicated in several psychiatric conditions but efforts to more clearly understand the mechanisms of this influence have been hampered by the complexity of this system at the receptor level. There are at least 14 distinct receptors that mediate the effects of 5-HT as well as several enzymes that control its synthesis and metabolism. Pharmacologic agents that target specific receptors have provided clues regarding the function of these receptors in the human brain. 5-HT is also an important modulator of neural development and several groups have employed a genetic strategy relevant to behavior. Several inactivation mutations of specific 5-HT receptors have been generated producing interesting behavioral phenotypes related to anxiety, depression, drug abuse, psychosis, and cognition. In many cases, knockout mice have been used to confirm what has already been suspected based on pharmacologic studies. In other instances, mutations have demonstrated new functions of serotonergic genes in development and behavior. PMID- 12907921 TI - Mouse genetic approaches to feeding regulation: serotonin 5-HT2C receptor mutant mice. AB - Neural mechanisms underlying the regulation of ingestive behavior and energy balance are well conserved among mammals. Many neural pathways, each reflecting the function of many genes, interact to regulate these processes. Systematic genetic perturbations are not feasible in humans--the examination of gene functions relevant to feeding regulation must be performed in other species. Many advances in this field have been made through molecular genetic studies of mice, the most genetically tractable of mammalian species. The relevance of mouse ingestive behavior to the mechanisms underlying the regulation of feeding in humans is discussed. Approaches for evaluating the contributions of genes to the regulation of energy balance and to the actions of anorectic drugs are described in the context of studies focused on a line of mice lacking the serotonin 5-HT2C receptor subtype. These animal display reduced responsiveness to serotonergic anorexic drugs and a late-onset obesity syndrome associated with features reminiscent of common forms of human obesity. Developmental studies of energy balance uncovered a novel age-dependent physiological process that may contribute generally to the predisposition of humans and other mammals to accumulate fat stores during "middle-age." These findings are presented to illustrate considerations in the use of mouse molecular genetic technologies to investigate genetic influences on ingestive behavior and energy balance. PMID- 12907922 TI - Substance P antagonists: meet the new drugs, same as the old drugs? Insights from transgenic animal models. AB - Antidepressants that primarily target the reuptake of monoamines have been highly successful treatments. However, therapies with these drugs still have several drawbacks, namely severe side effects, delays in the onset of action, and a significant percentage of non-responders. Recently, non-peptidic antagonists of the neurokinin 1 receptor, or substance P antagonists, have emerged as a novel class of drugs with antidepressant efficacy that is comparable to current drugs, but a potentially reduced side effect profile. This review summarizes the pre clinical evidence derived from pharmacological and transgenic animal studies that suggests an important role for the substance P/neurokinin 1 system in anxiety and depression. Also, potential mechanisms by which substance P antagonists may produce their therapeutic effects are discussed. PMID- 12907923 TI - Genetic approaches to the study of synaptic plasticity and memory storage. AB - Long-term memory is believed to depend on long-lasting changes in the strength of synaptic transmission known as synaptic plasticity. Understanding the molecular mechanisms of long-term synaptic plasticity is one of the principle goals of neuroscience. Among the most powerful tools being brought to bear on this question are genetically modified mice with changes in the expression or biological activity of genes thought to contribute to these processes. This article reviews how strains of mice with alterations in the cyclic adenosine monophosphate/protein kinase A/cyclic adenosine monophosphate-response element binding protein signaling pathway have advanced our understanding of the biological basis of learning and memory. PMID- 12907924 TI - Methylphenidate-induced obsessive-compulsive symptoms in an elderly man. AB - An 82-year-old man with treatment-resistant depression and early Alzheimer's disease was started on methylphenidate. Significant obsessive-compulsive behavior ensued but diminished over several weeks when methylphenidate was replaced by fluvoxamine. The patient had no prior psychiatric history, but he had a sister with obsessive-compulsive disorder. It appears that methylphenidate precipitated the patient's pathological behavior. PMID- 12907925 TI - No respect. PMID- 12907926 TI - Neurosensory testing of the medial calcaneal and medial plantar nerves in patients with plantar heel pain. AB - Eighty-two patients with a chief complaint of plantar heel pain were evaluated for sensory abnormalities within the cutaneous distribution of both the medial calcaneal nerve and the medial plantar nerve, using quantitative neurosensory testing with a pressure-specified sensory device. The results showed that 22.68% of the patients displayed isolated abnormal sensory function within the distribution of the medial calcaneal nerve, whereas 49.48% of the patients displayed abnormal function within the distribution of both the medial calcaneal and the medial plantar nerves. Thus, 72.17% of the patients displayed abnormal sensory function within the distribution of the medial calcaneal nerve. Statistical analysis of the results, using the Pearson chi-square statistic and odds ratio, indicated that a significant percentage of patients with plantar heel pain, even early in the clinical course of plantar heel pain, display abnormal sensibility within the branches of the posterior tibial nerve, and specifically, within the distribution of the medial calcaneal nerve (P <.0008) and the medial plantar nerve (P <.0001). PMID- 12907927 TI - Second metatarsophalangeal joint fusion: a new technique for crossover hammertoe deformity. A preliminary report. AB - The author reports on second metatarsophalangeal joint arthrodesis for the severe crossover hammertoe deformity. Eleven patients underwent an arthrodesis of the second metatarsophalangeal joint with a.062 Kirschner wire intramedullary fixation and a dorsal small bone staple and resection of the proximal phalangeal head of the second digit. Outcomes were retrospectively reviewed at average follow-up time of 19 months. A subjective patient satisfaction survey along with a clinical and radiographic evaluation was undertaken in the postoperative period. Ten of 11 patients scored good to excellent results based on the patient subjective survey at final follow-up. Ten of 11 patients obtained primary arthrodesis. One patient experienced an asymptomatic nonunion. This new technique appears to be reliable for the maintenance of long-term correction of a crossover second toe with arthrodesis of the second metatarsophalangeal joint. PMID- 12907928 TI - Treatment of lateral ankle instability with transfer of the extensor digitorum brevis muscle. AB - Ankle sprains are the most common injuries sustained during sports and physical exercise. Treatment is usually conservative because most of these injuries heal without consequence. However, some injuries may be followed by chronic lateral pain and instability, and surgical stabilization is sometimes necessary. In select cases, there is little or no residual ligament or scar tissue remaining for late reconstruction. Proximal transfer of the origin of the extensor digitorum brevis muscle as a substitute for deficient ligament tissue can be used in these difficult cases. During the years 1971 to 1992, 13 ankles in 10 patients underwent surgery using the proximal extensor digitorum brevis muscle transfer method. At follow-up, all the ankles manifested functional stability and were stable with no clinical drawer sign. All had a normal range of motion in the ankle joint but showed a desired decreased supination range of motion throughout the hindfoot and ankle. The functional Karlsson scores were 84.5 +/- 18.8 before injury, 26.4 +/- 18.7 before surgery, and 83.6 +/- 18.7 at follow-up. Thus, the extensor digitorum brevis muscle transfer seems to be a useful alternative method of long-term ankle stabilization in these difficult chronic case; the results correlate well with a few other studies using this method. PMID- 12907929 TI - Survival of patients with primary pedal melanoma. AB - Malignant melanoma is increasing at a rate faster than any other cancer in the United States. Location of the primary tumor on the foot is associated with poorer prognosis. This study evaluates a cohort of 148 patients with melanoma of the lower extremity (37 foot or ankle and 111 leg, knee, or thigh) diagnosed at a university medical center during a 32-year period. The mean follow-up for the foot/ankle patients was 44 months. The overall 5-year survival rate was 52% for patients with a primary melanoma of the foot/ankle compared with 84% for patients with a primary melanoma elsewhere on the lower extremity. Although the study period extended over 32 years, nearly 65% of the foot/ankle patients were diagnosed in the last decade. This increase is most likely because of the documented increase in incidence of melanoma in the United States and an increasing referral pattern at our institution. PMID- 12907930 TI - Reconstruction of the pediatric flexible planovalgus foot by using an Evans calcaneal osteotomy and augmentative medial split tibialis anterior tendon transfer. AB - Seventeen consecutive pediatric patients (34 feet) underwent reconstruction of flexible pes planovalgus deformity between 1994 and 1999. The average follow-up was 24.9 months (range, 8 to 48 months). All patients underwent a medial split tibialis anterior tendon transfer/tenodesis in combination with an Evans calcaneal-lengthening osteotomy, Kidner or modified Kidner procedure, and tendo Achilles lengthening. Various angular parameters were measured both preoperatively and postoperatively, and improvement in each was noted. The calcaneal pitch improved by an average of 26 degrees (85.88%), the lateral talometatarsal angle improved by an average of 26.4 degrees (96.15%), the lateral talocalcaneal angle improved by an average of 14.8 degrees (34.26%), the anteroposterior talometatarsal angle improved by an average of 21.9 degrees (81.92%), the anteroposterior talocalcaneal angle improved by an average of 15.9 degrees (37.95%), the cuboid abduction angle improved by an average of 13.8 degrees (87.34%), and the talonavicular coverage angle improved by an average of 26.7 degrees (94.68%). Postoperative improvement of all angular measurements was statistically significant at the 95% confidence level (P =.05). In addition, American Orthopedic Foot and Ankle Society hindfoot/ankle scoring was performed preoperatively, 3 months after the second foot surgery, and at the time of maximal follow-up. A preoperative average score of 68.59 improved to 85.76 at 3 months after the second foot surgery, and improved to 96.55 at the time of the last follow-up. All 17 patients were satisfied with their surgical result because all returned for correction of the contralateral foot. All patients indicated a uniform willingness to have these procedures performed again. PMID- 12907931 TI - Digital artery flaps for closure of soft tissue defects of the forefoot. AB - Reconstruction of recalcitrant soft tissue defects in the weightbearing surface of the forefoot can be achieved by using a neurovascular island flap. Island flaps, based on a pedicle from either the proper digital artery or the common digital artery, were used to provide supple and durable coverage. A retrospective analysis was performed on 12 patients who underwent a total of 15 digital artery flaps. There were 7 patients with neuropathic ulcers, 7 with a dysfunctional scar, and 1 with an ischemic ulcer after lower-extremity bypass. There was a failure rate of 13%; 2 flaps fully necrosed, necessitating a revisional digital artery flap. Minor complications were reported in 73% of cases; average time to complete healing was 71 days. All healed flaps have remained viable and durable at an average follow-up of 22.5 months from the date of surgery (range, 3 to 61 months). PMID- 12907932 TI - Peroneal tendon injuries: an evaluation of 49 tears in 41 patients. AB - Forty-one patients sustaining 49 tears of the peroneal tendons were evaluated prospectively a minimum of 1 year after surgical treatment. Preoperative and postoperative function and activity were assessed by using the American Orthopedic Foot and Ankle (AOFAS) score. Mean age at the time of surgery was 44.0 +/- 11.7 years. Mean follow-up after the index surgery was 35.5 +/- 22.2 months. There were 17 women and 24 men. One woman had bilateral surgery 1 year apart. There were a total of 18 tears of the peroneus longus tendon; 11 of these were isolated, whereas 7 had a combined tear with peroneus brevis. There 31 peroneus brevis tears; 24 of these were isolated and 7 were combined. Using 3-way analysis of variance, there were no significant differences in return to activity or postoperative AOFAS scores among those with a longus, brevis, or combined tear. The mean return to activity for peroneus longus, peroneus brevis, and combined tears were 3.2, 3.6, and 3.7 months, respectively. The mean postoperative AOFAS scores were 90.6, 90.8, and 84.3 respectively. The mean preoperative AOFAS score was 52.0 +/- 16.8. The mean postoperative score was 89.7 +/- 10.3 (P <.00001). Using this scoring system, there were 24 excellent, 12 good, 4 fair, and 2 poor scores. Three patients underwent additional surgery. Fourteen of 16 athletes returned to full sporting level. The average return to activity for the entire group was 3.49 +/- 1.15 months. PMID- 12907933 TI - Forensic implications of the foot and ankle. AB - To clinicians, forensic medicine and the foot and ankle complex may appear to have little connection. However, if decomposed, burned, fragmented, or skeletonized human remains are recovered, the foot and ankle can provide information leading to identification and incident analysis. Furthermore, analysis of data collected at accidents can lead to safety modifications. This article reviews common elements of forensic medicine applied to the foot and ankle in 2 areas: establishing positive identification by using radiographic comparisons and analyzing aviation incidents. An understanding of these cases will enhance clinicians' awareness of the potential forensic value of the foot and ankle. PMID- 12907935 TI - Free os cuboideum secundarium: a case report. AB - The free os cuboideum secundarium is one of the most rare accessory bones of the foot and is located proximal and plantar to the cuboid. In a 9-year-old boy, a free os cuboideum secundarium of the left foot was resected for persistent pain after trauma. Histologic examination showed degenerative changes of the hyaline cartilage that covered the entire bone. The central spongiosa appeared normal. The patient is free of pain and is still playing football 9.5 years after resection of the accessory bone. Follow-up examination showed no clinical sign of pathology except a soft scar with little discomfort when walking without shoes. Radiographs showed complete remodeling of left cuboid and calcaneus with normal bony configuration of both feet. PMID- 12907934 TI - Granuloma formation after chevron osteotomy fixation with absorbable copolymer pin: a case report. AB - After fixation of a chevron osteotomy for hallux valgus of the left foot with a bioabsorbable copolymer, a delayed onset of pain at the surgical site developed. Magnetic resonance imaging showed a cystic lesion that was unresponsive to conservative treatment. Surgical excision determined the lesion to be a giant cell granuloma formed as a foreign body response to the degrading copolymer. PMID- 12907936 TI - Ankle swelling in a 6-year-old boy with unusual presentation: report of a rare case. AB - A 6-year-old boy with a history of pain and swelling in the right ankle was referred to our medical center. A cascading algorithm helped us to better evaluate this case of an osteochondroma of talus. The diagnosis was based on clinical and radiologic findings and on histopathologic analysis of the excised tissue. Although osteochondroma, or osteocartilaginous exostosis, is a common skeletal neoplasm, it is rarely seen in the foot. The rare location, a young age group, and an inexperienced surgeon may make the diagnosis confusing. We presented a case that shows the importance of an orderly evaluation of a musculoskeletal neoplasm. This may be helpful for the physician to better understand the clinical implications of any case before attempting a surgical intervention. PMID- 12907937 TI - Use of a hybrid "kickstand" external fixator for pressure relief after soft tissue reconstruction of heel defects. PMID- 12907938 TI - The Rousso stitch: a new everting skin closure technique. AB - The purpose of this article is to describe a new suturing technique that everts the skin and is a modification the horizontal mattress suture called the Rousso stitch. The technique is described step by step, along with reference pictures. The Rousso stitch technique was developed based on the well-documented theory behind everting the edges of thicker skin on closure of surgical incisions to allow for ideal coaptation and minimal scar formation. It is an easy technique to learn and appears to be a good time-saving option for the surgeon to use for the same applications of the typical horizontal mattress suture. PMID- 12907939 TI - Long oblique distal osteotomy of the fifth metatarsal for correction of tailor's bunion: a retrospective review. PMID- 12907940 TI - Modified Kalish osteotomy: a simple approach to minimizing proximal dorsal stress risers. PMID- 12907941 TI - The Mott-Hubbard insulating state and orbital degeneracy in the superconducting C60(3-) fulleride family. AB - Electron correlation controls the properties of important materials such as superconducting and magnetoresistive transition metal oxides and heavy fermion systems. The role of correlation in driving metal-to-insulator transitions assumes further importance because many superconducting materials are located close to such transitions. The nature of the insulating ground state often reveals the dominant interactions in the superconductor, as shown by the importance of the properties of La2CuO4 in understanding the high-temperature superconducting cuprates. The A3C60 alkali metal fullerides are superconducting systems in which the role of correlation in both the normal state and the superconducting pairing mechanism is controversial, because no magnetic insulator comparable to the superconducting materials has been identified. We describe the first example of a cubic C60(3-) system with degenerate orbitals that adopts the Mott-Hubbard insulating localized electron ground state. Electron repulsion is identified as the interaction that is suppressed on the transition to metallic and superconducting behaviour in the fullerides. This observation is combined with ab initio calculations to demonstrate that it is the orbital degeneracy that allows the superconducting cubic A3C60 fullerides to remain metallic while provoking electron localization in systems with lower symmetry. PMID- 12907942 TI - Nanoporous carbide-derived carbon with tunable pore size. AB - Porous solids are of great technological importance due to their ability to interact with gases and liquids not only at the surface, but throughout their bulk. Although large pores can be produced and well controlled in a variety of materials, nanopores in the range of 2 nm and below (micropores, according to IUPAC classification) are usually achieved only in carbons or zeolites. To date, major efforts in the field of porous materials have been directed towards control of the size, shape and uniformity of the pores. Here we demonstrate that porosity of carbide-derived carbons (CDCs) can be tuned with subangstrom accuracy in a wide range by controlling the chlorination temperature. CDC produced from Ti3SiC2 has a narrower pore-size distribution than single-wall carbon nanotubes or activated carbons; its pore-size distribution is comparable to that of zeolites. CDCs are produced at temperatures from 200-1,200 degrees C as a powder, a coating, a membrane or parts with near-final shapes, with or without mesopores. They can find applications in molecular sieves, gas storage, catalysts, adsorbents, battery electrodes, supercapacitors, water/air filters and medical devices. PMID- 12907943 TI - The endoplasmic reticulum is the site of cholesterol-induced cytotoxicity in macrophages. AB - Excess cellular cholesterol induces apoptosis in macrophages, an event likely to promote progression of atherosclerosis. The cellular mechanism of cholesterol induced apoptosis is unknown but had previously been thought to involve the plasma membrane. Here we report that the unfolded protein response (UPR) in the endoplasmic reticulum is activated in cholesterol-loaded macrophages, resulting in expression of the cell death effector CHOP. Cholesterol loading depletes endoplasmic reticulum calcium stores, an event known to induce the UPR. Furthermore, endoplasmic reticulum calcium depletion, the UPR, caspase-3 activation and apoptosis are markedly inhibited by selective inhibition of cholesterol trafficking to the endoplasmic reticulum, and Chop-/- macrophages are protected from cholesterol-induced apoptosis. We propose that cholesterol trafficking to endoplasmic reticulum membranes, resulting in activation of the CHOP arm of the UPR, is the key signalling step in cholesterol-induced apoptosis in macrophages. PMID- 12907944 TI - Hypoxia-inducible VEGF gene delivery to ischemic myocardium using water-soluble lipopolymer. AB - Therapeutic angiogenesis with gene encoding vascular endothelial growth factor (VEGF) is a new potential treatment in cardiovascular disease. However, unregulated VEGF-mediated angiogenesis has the potential to promote tumor growth, accelerate diabetic proliferative retinopathy, and promote rupture of atherosclerotic plaque. To be safe and effective, gene therapy with VEGF must be regulated. To limit the risk of pathological angiogenesis, we developed a hypoxia inducible VEGF gene therapy system using the erythropoietin (Epo) enhancer and water-soluble lipopolymer (WSLP). pEpo-SV-VEGF or pSV-VEGF-Epo was constructed by insertion of the Epo enhancer upstream of the Simian Virus 40 (SV40) promoter or downstream of the poly(A) signal of pSV-VEGF. In vitro transfection showed that pEpo-SV-VEGF, not pSV-VEGF-Epo, induced the VEGF expression in hypoxic cells. In addition, the VEGF protein, which was produced from the Epo-SV-VEGF-transfected and hypoxia-incubated cells, was able to enhance the proliferation of the endothelial cells. Injection of the pEpo-SV-VEGF/WSLP complex showed that the expression of VEGF was induced in ischemic myocardium, compared to normal myo cardium. Therefore, with the localized induction of VEGF and the low cytotoxicity of WSLP, the pEpo-SV-VEGF/WSLP system may be helpful to eventually treat ischemic heart disease. PMID- 12907945 TI - Protection against collagen-induced arthritis by intramuscular gene therapy with an expression plasmid for the interleukin-1 receptor antagonist. AB - The interleukin-1 receptor antagonist (IL-1Ra) is an endogenous protein that can prevent the binding of IL-1 to its cell-surface receptors. Among a number of techniques for gene transfer in vivo, the direct injection of naked DNA into muscle is simple, inexpensive and safe. In this study, we evaluated the potential of intramuscular gene therapy with plasmid DNA containing the cDNA for IL-1Ra in the prevention of murine collagen-induced arthritis (CIA). DBA/1 mice were immunized with bovine type II collagen. At 4 weeks after the initial immunization, expression plasmid for IL-1Ra was injected into four selected sites in the thigh and calf muscles of DBA/1 mice. Control mice received the same plasmid, but lacking the IL-1Ra coding sequence. Macroscopic analysis of paws for redness, swelling and deformities showed that the onset of moderate to severe CIA in the paws of mice injected with IL-1Ra DNA was significantly prevented (P<0.05). In addition, both the synovitis and the cartilage erosion in knee joints were dramatically reduced in mice treated with IL-1Ra DNA (P<0.05). The expression of IL-1beta was significantly decreased in the ankle joints of mice treated with IL-1Ra (P<0.01). Interestingly, the levels of IL-1Ra in sera and joints after intramuscular injection of IL-1Ra DNA were significantly lower than when protein had been used in previous reports, suggesting that the therapeutic effect may be achieved by an alternative mechanism(s) rather than by systemic elevation of IL-1Ra. These observations provide the first evidence that direct intramuscular injection of expression plasmid for IL-1Ra may effectively suppress the inflammatory pathology in arthritis. PMID- 12907946 TI - Targeting recombinant adeno-associated virus vectors to enhance gene transfer to pancreatic islets and liver. AB - Human pancreatic islet cells and hepatocytes represent the two most likely target cells for genetic therapy of type I diabetes. However, limits to the efficiency of rAAV serotype 2 (rAAV2)-mediated gene transfer have been reported for both of these cell targets. Here we report that nonserotype 2 AAV capsids can mediate more efficient transduction of islet cells, with AAV1 being the most efficient serotype in murine islets, suggesting that receptor abundance could be limiting. In order to test this, we generated rAAV particles that display a ligand (ApoE) that targets the low-density lipoprotein receptor, which is present on both of these cell types. The rAAV/ApoE viruses greatly enhanced the efficiency of transduction of both islet cells ex vivo and murine hepatocytes in vivo when compared to native rAAV2 serotype (220- and four-fold, respectively). The use of receptor-targeted rAAV particles may circumvent the lower abundance of receptors on certain nonpermissive cell types. PMID- 12907948 TI - Specific hammerhead ribozymes reduce synthesis of cation-independent mannose 6 phosphate receptor mRNA and protein. AB - Storage diseases because of lysosomal enzyme deficiencies may be treated by the transplantation of cells that secrete the enzyme which is deficient in patients. One can expect that increasing the amount of secreted enzymes will improve the therapy efficacy. Secretion of lysosomal enzymes can be enhanced by reducing the mannose 6-phosphate receptor involved in the lysosomal sorting of newly synthesized lysosomal enzymes. For this purpose, we have constructed hammerhead ribozymes targeting the mRNA of the large murine mannose 6-phosphate receptor (M6PR300). In vitro ribozymes cleave M6PR300 RNA fragments efficiently with cleavage rates of 69-93% after 3 h of incubation. Ribozymes were cloned into an expression vector in which they are integrated into the VaI adenovirus RNA to increase stability and in which they are transcribed from an RNA polymerase III promoter. These plasmids were transiently transfected into BHK cells to investigate in vivo activity. Two ribozymes reduce efficiently the levels of murine M6PR300 mRNA in transient transfection experiments to 42-45%. This correlates with the reduction of M6PR300 biosynthesis, which is reduced also to 37% of normal. We can also demonstrate that the reduction in M6PR300 is sufficient to increase a lysosomal enzyme secretion. PMID- 12907947 TI - Hepatocyte growth factor gene transfer into the liver via the portal vein using electroporation attenuates rat liver cirrhosis. AB - Although a variety of gene transfer methods to the liver have been designed, there are some problems such as the transfection efficiency and safety. In the present study, we developed a modified method of gene transfer into the liver by infusion of plasmid DNA via the portal vein followed by electroporation. After green fluorescence protein gene transfer, transgene expressions were detected in 24 h, and then maximally at 3 days, and persisted for 3 weeks. Histological analysis revealed that very mild tissue damage was induced in the liver to which electroporation was applied. In the second study, human hepatocyte growth factor (HGF) was more detected in the liver injected with 500 microg of human HGF gene than 100 microg of human HGF gene. However, serum HGF did not increase with 100 or 500 microg of human HGF gene. Moreover, 500 microg of HGF gene transfer into the liver by using this method could achieve the long survival of all dimethylnitrosamine-treated rats and attenuate the fibrous regions in the liver. These results suggest that HGF gene transfer into the liver via the portal vein using electroporation might be one of the useful methods for the treatment of various liver diseases. PMID- 12907949 TI - PEI - a potent, but not harmless, mucosal immuno-stimulator of mixed T-helper cell response and FasL-mediated cell death in mice. AB - Polyethyleneimine (PEI) is one of the most effective gene delivery systems available today. However, very little is known about its ability to stimulate a systemic immune response and the molecular mechanisms thereof. However, this information is vital for the future development of new gene delivery systems. Here we address this issue by studying gene expression profiles from spleen lymphocytes after in vivo immunization of mice with PEI formulated with a reporter plasmid (PEI+) or the formulation alone (PEI-). PEI- was found to provoke the activation of genes with important immunostimulatory functions, but without the necessary costimulatory signals. PEI+ resulted in: a mixed Th1/Th2 response; activation of both CD8(+) and CD4(+) T cells, with a larger effect on CD4(+); and FasL-mediated antigen-induced cell death. A comparison of the immune responses of PEI+ with that of the clinically used tetanus toxoid-aluminum phosphate vaccine showed that the DNA vaccine provoked a stronger immune response as compared to the protein vaccine. However, many genes involved in other cellular responses such as apoptosis, stress responses and oncogenesis were activated in PEI+, supporting the theory of immunostimulation by danger genes, but also pointing toward possible adverse reactions such as Alzheimer's disease. PMID- 12907950 TI - A comparison of gene repair strategies in cell culture using a lacZ reporter system. AB - Synthetic oligonucleotides and DNA fragments of less than 1 kilobase (kb) have been shown to cause site-specific genetic alterations in mammalian cells in culture and in vivo. We have used a lacZ reporter gene system to compare the efficiency of episomal and chromosomal gene repair in human embryonic kidney epithelial cells (HEK293), Chinese Hamster Ovary fibroblasts (CHOK1), human bronchial epithelial cells (16HBE), and mouse embryonic stem (ES) cells. The lacZ gene contains a G to A nucleotide change, (Glu to Lys mutation) that abrogates beta-galactosidase activity. We compared the efficiency of different gene repair methods to correct this mutation and restore beta-galactosidase activity. We evaluated PCR-generated double-stranded DNA fragments of 0.52-1.9 kb, single stranded DNA oligonucleotides of 20, 35, or 80 bases containing internal phosphorothioate links, and a 68 base RNA:DNA oligonucleotide. All of the oligonucleotides and DNA fragments showed some gene repair ability with an episomal plasmid. Short DNA fragments of 0.52 kb or greater gave the highest frequencies of episomal gene repair while single-stranded DNA oligonucleotides gave the highest frequency of chromosomal repair. In the context of a chromosomal target, antisense DNA oligonucleotides gave 5-fold higher frequencies of gene repair than their sense counterparts. The RNA:DNA chimeric oligonucleotide gave little or no gene repair on either a chromosomal or episomal target. PMID- 12907951 TI - Therapeutic effect of DNA vaccines combined with chemotherapy in a latent infection model after aerosol infection of mice with Mycobacterium tuberculosis. AB - The prevention of Mycobacterium tuberculosis (M. tuberculosis) reactivation would greatly reduce the incidence of the disease, particularly among the elderly. Here, we evaluated the efficacy of DNA vaccine in combination with a conventional TB chemotherapy on the prevention of M. tuberculosis reactivation. Mice were treated with isoniazid and pyrazinamide for 3 months from 4 weeks after aerosol infection with M. tuberculosis H37Rv. During this period of chemotherapy, DNA immunization was performed three times monthly with an antigen 85A (Ag85A) DNA or an IL-12 mutant (IL-12N220L) DNA, which is known to lead to a reduction in the secretion of the p40 subunit, but not of a bioactive IL-12p70. The reactivation of M. tuberculosis was dramatically reduced in mice treated with either Ag85A DNA (P<0.01) or IL-12N220L DNA (P<0.05) in combination with chemotherapy, compared with control mice receiving only chemotherapy. Ag85A DNA vaccine showed higher IFN-gamma responses to Ag85A protein, but a lower response to culture filtrate than IL-12N220L DNA vaccine. In addition, Ag85A DNA vaccine prevented the reactivation of M. tuberculosis more efficiently than IL-12N220L DNA vaccine, indicating that Ag85A-specific IFN-gamma response might correlate with M. tuberculosis control. This study suggests that immunotherapy using Ag85A or IL 12N220L DNA vaccine combined with conventional chemotherapy might be effective clinically for the prevention of tuberculosis reactivation and may offer a more effective cure for humans than chemotherapy alone. PMID- 12907952 TI - Focused ultrasound (HIFU) induces localized enhancement of reporter gene expression in rabbit carotid artery. AB - The development of accurate, safe, and efficient gene delivery remains a major challenge towards the realization of gene therapeutic prevention and treatment of cardiovascular diseases. In this study, we investigated the ability of high intensity focused ultrasound (HIFU), a form of mechanical wave transmission, to act as a noninvasive tool for the enhancement of in vivo gene transfer into rabbit carotid arteries. Segments of the common carotid arteries of New Zealand white rabbits were isolated and infused with plasmid DNA encoding the reporter beta-galactosidase either with or without the addition of ultrasound contrast agent consisting of small (approximately 2-5 microm) gas-filled human albumin microspheres to augment cavitation. Infused arteries were exposed to pulsed ultrasound for 1 min (frequency 0.85 MHz, burst length 50 ms, repetition frequency 1 Hz, duration 60 s, peak pressure amplitude of 15 MPa). At 6.3 MPa, HIFU enhanced gene expression eight-fold, and 17.5-fold in the presence of contrast. We found increasing amounts of beta-galactosidase expression in the carotid vessel with increasing pressure amplitude. This dose-response relation was present with and without contrast. Without contrast, no vessel damage was detected up to 15 MPa, while the addition of contrast induced side effects above a threshold of 6.3 MPa peak pressure. The entire procedure was feasible and safe for the animals, and the results suggest that HIFU has the potential to assist in the noninvasive spatial regulation of gene transfer into the vascular system. PMID- 12907953 TI - Electroporation as a method for high-level nonviral gene transfer to the lung. AB - To increase the levels of pulmonary gene transfer by nonviral vectors, we have adopted electroporation protocols for use in the lung. A volume of 100-200 microl of purified plasmid DNA suspended in saline was instilled into the lungs of anesthetized mice. Plasmids expressed luciferase, or beta-galactosidase under control of the CMV immediate-early promoter and enhancer. Immediately following delivery, a series of eight square wave electric pulses of 10 ms duration each at an optimal field strength of 200 V/cm were administered to the animals using 10 mm Tweezertrodes (Genetronics, San Diego, CA, USA). The electrodes were placed on either side of the chest, which had been wetted with 70% ethanol. The animals recovered and survived with no apparent trauma until the experiments were terminated at the desired times, between 1 and 7 days post-treatment. Gene expression was detected by 1 day postelectroporation and peaked between 2 and 5 days. By 7 days, expression was back to baseline. By contrast, essentially no gene expression was detected in the absence of electric pulses. Using a beta galactosidase-expressing plasmid, the distribution of gene expression appeared to be concentrated in the periphery of the lung, but was also present throughout the parenchyma. The primary cell types expressing gene product include alveolar type I and type II epithelial cells. No inflammation or lung injury was detected histologically or by cytokine measurements in lungs at either 1 or 24 h following electroporation treatment. These results provide evidence that electroporation is a safe and effective means for introducing naked DNA into the lung and form the basis for future studies on targeted pulmonary gene therapy. PMID- 12907954 TI - A novel combination of promoter and enhancers increases transgene expression in vascular smooth muscle cells in vitro and coronary arteries in vivo after adenovirus-mediated gene transfer. AB - Recombinant adenoviruses are employed widely for vascular gene transfer. Vascular smooth muscle cells (SMCs) are a relatively poor target for transgene expression after adenovirus-mediated gene delivery, however, even when expression is regulated by powerful, constitutive viral promoters. The major immediate-early murine cytomegalovirus enhancer/promoter (MIEmCMV) elicits substantially greater transgene expression than the human cytomegalovirus promoter (MIEhCMV) in all cell types in which they have been compared. The Woodchuck hepatitis virus post transcriptional regulatory element (WPRE) increases transgene expression in numerous cell lines, and fragments of the smooth muscle myosin heavy chain (SMMHC) promoter increase expression within SMC from heterologous promoters. We therefore, compared the expression of beta-galactosidase after adenovirus mediated gene transfer of lacZ under the transcriptional regulation of a variety of combinations of the promoters and enhancers described, in vitro and in porcine coronary arteries. We demonstrate that inclusion of WPRE and a fragment of the rabbit SMMHC promoter along with MIEmCMV increases beta-galactosidase expression 90-fold in SMC in vitro and approximately 40-fold in coronary arteries, compared with vectors in which expression is regulated by MIEhCMV alone. Expression cassette modification represents a simple method of improving adenovirus-mediated vascular gene transfer efficiency and has important implications for the development of efficient cardiovascular gene therapy strategies. PMID- 12907955 TI - Coxsackievirus and adenovirus receptor (CAR)-positive immature osteoblasts as targets of adenovirus-mediated gene transfer for fracture healing. AB - Adenovirus vectors are expected to be a powerful tool for gene therapy to treat severe fractures. Adenovirus invades cells through binding to the coxsackievirus and adenovirus receptor (CAR) on the cell membrane. CAR expression is low in normal adult animals, but it is induced on regenerating cells in some experimental models. We made a rib fracture model in mice and evaluated the histological changes and CAR mRNA expression by RT-PCR 1, 5, 10, 14, and 21 days after the fracture. CAR mRNA was expressed exclusively in the fractured ribs at each time point, but not in the normal ribs. We detected the CAR protein immunohistochemically in fibroblast-like cells in the fracture callus on days 10 and 14 after fracture. In situ hybridization showed that these fibroblast-like cells expressed mRNA of type I collagen and osteopontin, but not osteocalcin, defining the cells as immature osteoblasts. We then transferred small doses (10(4)-10(8) PFU) of lacZ-expressing adenovirus vector into immature osteoblasts on day 14. beta-galactosidase was detected only on the immature osteoblasts at every dose. Immature osteoblasts play an important role in the matrix replacement step in fracture healing. CAR-mediated gene transfer into immature osteoblasts can be reasonable for adenovirus-mediated treatment of fracture healing. PMID- 12907956 TI - Twice a day? PMID- 12907957 TI - Multi-speciality agreement needed. PMID- 12907958 TI - Tooth notation minus confusion. PMID- 12907960 TI - Prolonged paraesthesia. PMID- 12907973 TI - In our opinion: The First Five Years--A framework for Dental Undergraduate Education. PMID- 12907974 TI - General medicine and surgery for dental practitioners. Part 6: The endocrine system. PMID- 12907975 TI - Prevention. Part 4: Toothbrushing: what advice should be given to patients? PMID- 12907983 TI - Clinical characteristics of somatization in dental practice. AB - OBJECTIVES: This study was a first attempt to derive an estimate of the likely incidence of somatization-specific behaviour in a dental setting and its relationship with both somatic symptoms and symptoms of depression. METHODS: Somatization-specific behaviour was operationalized as reporting of inexplicable dental symptoms (eg pain), remarkable frequent attendance at a dental surgery, inexplicably high treatment use or unreasonable requests with regard to treatment. RESULTS: Of the 309 patients surveyed, 8.7%o fulfilled one or more of the criteria for somatization-specific behaviour. This was particularly manifested by a high attendance rate (6.8%). Women exhibited somatization specific behaviour significantly more often (73%) than men (27%9). Support was found for the hypothesis that individuals exhibiting characteristics of somatization-specific behaviour would present themselves to dentists more often with dental complaints and would suffer more from symptoms of depression than patients that did not display these characteristics. CONCLUSIONS: The results suggest that somatization is a factor in dental practice. PMID- 12907999 TI - An outbreak of Norwalk-like virus gastroenteritis in an aged-care residential hostel. PMID- 12907985 TI - Fluoride supplements and changes in tooth decay on the Island of Tristan da Cunha: 1966-1996. AB - BACKGROUND: The island of Tristan Da Cunha is one of the few examples in the world of a remote enclosed community. The inhabitants of the island were transported to England in 1961 as a result of a volcanic eruption and during their time in England they received a detailed dental health examination. They were later examined back on the island in 1966 by Dr John Fisher and in 1982 a school fluoride supplementation programme was introduced. AIM: The present paper reports the results of the first dental health survey to be carried out on the island since the introduction of a regular school fluoride supplementation programme in 1982. SUBJECTS AND METHODS: A cohort of 6-19-year-old subjects on the island were examined in 1996 using a similar protocol to that which was used by Fisher in the 1966 examination. RESULTS: Comparing the 6-12-year-olds who were caries free in 1966 with those caries free in 1996 using the Chi-squared statistic reveals a statistically significant greater number caries free in the more recent cohort (X2 - 6.0, P- 0.014). For the older age group (13-19 years), a similar comparison reveals a highly statistically significant difference(Z2 - 12.26, P 0.005). CONCLUSIONS: A significant increase in the number of caries free 6-19-year-old children in Tristan Da Cunha between 1966 and 1996 was noted: 1) This is a significant finding in the light of the school fluoride supplementation programme that was introduced on the island in 1982; and 2) Appropriate fluoride supplementation regimes may have conferred a protective effect in a group of children with a cariogenic diet. The paper discusses the significance of this study in the support of fluoridation as a method of reducing the prevalence of dental caries a nd also discusses possibilities for future research on the island of Tristan Da Cunha. PMID- 12907984 TI - Retention of prefabricated and individually cast root canal posts in vitro. AB - OBJECTIVES: The aim of the study was to evaluate the retention of prefabricated root canal posts made of a variety of materials that have recently been introduced into dentistry. MATERIALS AND METHODS: The posts studied were CosmoPost, Composipost carbon fibres, Composipost Aestheti-Plus, Composipost Light-Post and Para Post Fiber White. The posts were luted in extracted human premolars and the cores were built up with the resin composites recommended by the manufacturers. The retention of individually cast gold alloy posts luted with zinc-phosphate cement were used as reference. A universal testing machine was used to determine the retention of each cemented post Data were compared using ANOVA supplemented with Fisher's PLSD at a significance level of p < 0.05. RESULTS: Only the CosmoPost system exhibited retention values that were significantly lower than for the conventionally cast gold alloy posts luted with zinc-phosphate cement The force necessary to loosen the CosmoPost specimens was significantly less than that needed to loosen the Composipost. Estheti-Plus (p < 0.05) and the Composipost Ught-Post systems (p < 0.001]. The force necessary to loosen the ParaPost Fiber White specimens was significantly less than for the Composipost Light-Post system (p < 0.01). Other combinations did not differ significantly (p > 0.05). CONCLUSIONS: When zirconium oxide ceramic posts are luted with resin composites the bonding between the ceramic and resin composite seems to be weak. Further studies are thus necessary to achieve improved retention of all-ceramic root canal posts. PMID- 12908000 TI - Firework injuries in New South Wales, 1992-93 to 2001-02. PMID- 12908001 TI - An innovative sexual health medication order system for the Far West Area Health Service. PMID- 12908002 TI - A consumer-initiated survey of women's experiences of breast cancer services in New South Wales. PMID- 12908003 TI - Release of the 2002 Area Health Service Reports. PMID- 12908004 TI - Release of New South Wales Mothers and Babies 2001. PMID- 12908005 TI - Methicillin resistant Staphylococcus aureus. PMID- 12908006 TI - Communicable Diseases Report, NSW: June 2003. PMID- 12908007 TI - Effects of anxiolytic drugs on rewarding and aversive behaviors induced by intracranial stimulation. AB - In considering the characteristics of the action of anxiolytic drugs and their mechanism in the brain, it may be necessary not only to study the behavioral pharmacology but also to perform brain site research. In the present study, the action of anxiolytic drugs was examined with respect to various behaviors that were induced by stimulating the brain areas related to emotions such as reward (pleasure) or aversion in rats. First, the low rate of response in lateral hypothalamic self-stimulation behavior was induced by schedules of low current brain stimulation, variable interval (VI) and differential reinforcement of low rate (DRL). Anxiolytic drugs such as benzodiazepines facilitated these low-rate responses. The drug susceptibility was highest in the low current stimulation, lower in the VI stimulation, and lowest in the DRL stimulation schedules. Furthermore, it was found by the auto-titration method in intracranial self stimulation behavior that anxiolytic drugs decreased the threshold of stimulation reward. Second, it was recognized using the decremental lever pressing (DLP) paradigm that anxiolytic drugs increased the threshold of aversive stimulation of mesencephalic dorsal central gray (DCG), and this increasing effect of the drug was antagonized by GABA receptor blockers such as biccuculline. Finally, it was examined whether or not the conflict situation is established by combining brain stimulation reward and aversion, such as foot-shock or DCG stimulation. As a result, the conflict behavior was established by combining not only the brain stimulation reward and foot-shock aversion, but also the brain stimulation reward and DCG stimulation aversion. Further anxiolytic drugs exhibited anti-conflict action in both situations. The susceptibility of anxiolytic drugs was higher with respect to the conflict behavior induced by intracranial reward and aversion than to that induced by the conventional method based on milk reward and foot-shock aversion. These results suggest that behavioral methods using brain stimulation can examine the mechanisms of direct drug action at the brain stimulation site. Indeed, in the present brain stimulation behavioral study, anxiolytic drugs such as benzodiazepines increased the stimulation threshold in lateral hypothalamic self-stimulation and inhibited the DCG aversive stimulation, thus resulting in an anticonflict action of the drugs. PMID- 12908008 TI - Genome-wide search for strabismus susceptibility loci. AB - The purpose of this study was to search for chromosomal susceptibility loci for comitant strabismus. Genomic DNA was isolated from 10mL blood taken from each member of 30 nuclear families in which 2 or more siblings are affected by either esotropia or exotropia. A genome-wide search was performed with amplification by polymerase chain reaction of 400 markers in microsatellite regions with approximately 10 cM resolution. For each locus, non-parametric affected sib-pair analysis and non-parametric linkage analysis for multiple pedigrees (Genehunter software, http://linkage.rockefeller.edu/soft/) were used to calculate multipoint lod scores and non-parametric linkage (NPL) scores, respectively. In sib-pair analysis, lod scores showed basically flat lines with several peaks of 0.25 on all chromosomes. In non-parametric linkage analysis for multiple pedigrees, NPL scores showed one peak as high as 1.34 on chromosomes 1, 2, 4, 7, 10, 15, and 16, while 2 such peaks were found on chromosomes 3, 9, 11, 12, 18, and 20. Non parametric linkage analysis for multiple pedigrees of 30 families with comitant strabismus suggested a number of chromosomal susceptibility loci. Our ongoing study involving a larger number of families will refine the accuracy of statistical analysis to pinpoint susceptibility loci for comitant strabismus. PMID- 12908010 TI - Radiation damage to the normal monkey brain: experimental study induced by interstitial irradiation. AB - Radiation damage to normal brain tissue induced by interstitial irradiation with iridium-192 seeds was sequentially evaluated by computed tomography (CT), magnetic resonance imaging (MRI), and histological examination. This study was carried out in 14 mature Japanese monkeys. The experimental area received more than 200-260 Gy of irradiation developed coagulative necrosis. Infiltration of macrophages to the periphery of the necrotic area was seen. In addition, neovascularization, hyalinization of vascular walls, and gliosis were found in the periphery of the area invaded by the macrophages. All sites at which the vascular walls were found to have acute stage fibrinoid necrosis eventually developed coagulative necrosis. The focus of necrosis was detected by MRI starting 1 week after the end of radiation treatment, and the size of the necrotic area did not change for 6 months. The peripheral areas showed clear ring enhancement with contrast material. Edema surrounding the lesions was the most significant 1 week after radiation and was reduced to a minimum level 1 month later. However, the edema then expanded once again and was sustained for as long as 6 months. CT did not provide as clear of a presentation as MRI, but it did reveal similar findings for the most part, and depicted calcification in the necrotic area. This experimental model is considered useful for conducting basic research on brachytherapy, as well as for achieving a better understanding of delayed radiation necrosis. PMID- 12908009 TI - Effect of ornithine on the ileal histology, nitric oxide production and lipid peroxidation in LPS-induced endotoxemia. AB - Effect of ornithine which is known to inhibit L-arginine uptake via cationic amino acid transport system has been tested, and compared to aminoguanidine, an iNOS inhibitor in lypopolysaccharide (LPS)-induced endotoxemia in rats. Serum nitrite/nitrate and malondialdehyde (MDA) level have been measured, and ileal histology has also been examined. Endotoxin increased serum nitrite/nitrate and MDA levels from 15.7+/- 2.4 micromol/ml and 2.1 +/-0.2 nmol/ml to 23.1 +/- 1.0 micromol/ml and 5.2+/- 0.3 nmol/ml (both P<0.05), respectively. In addition, LPS caused ileal degeneration. L-ornithine (500 mg/kg) did not improve septic manifestations, i.e., serum nitrite/nitrate and MDA levels did not differ from those in endotoxemia. Neither does it have an improving action on ileal histology. However, higher dose of L-ornithine (2,500 mg/kg) lowered the increased level of nitrite/nitrate and MDA by LPS. Moreover, it restored ileal histology from grade 3 (median) to 0 (median) (P<0.05). On the other hand, aminoguanidine (100 mg/kg) normalized serum nitrite/nitrate and MDA levels but not ileal histology in endotoxemic rats. In conclusion, high dose of L-ornithine could improve endotoxemic parameters in LPS-treated rats. PMID- 12908011 TI - Partial liquid ventilation does not affect BALF TNF-, MIP-2, CINC-1 concentrations, or CD11b cell surface expression, but does increase macrophage proportion among BALF cells in the acute phase of rat LPS-induced lung injury. AB - To elucidate the mechanism of anti-inflammatory effect of partial liquid ventilation (PLV), cytokine concentration, surface CD11b, and macrophage expression were investigated in BALF. The 30-minutes group was treated with gas ventilation (GV) for 30 minutes after intratracheal LPS administration. The GV group was prepared in the same manner as the 30-minutes group, then the GV was continued for the following 2 hours. The PLV group was treated in the same manner as the 30-minutes group, and then received PLV with perflubron for the following 2 hours. Animals were euthanized to receive BAL. The PLV group showed a tendency to have a higher concentration than the GV group of TNF-alpha, MIP-2, and CINC-1 as measured by ELISA, although there were no significant differences. The ratio of expressions of CD11b and macrophages to total leukocytes were determined by flow-cytometry. There were no significant differences in the ratio of CD11b positive expression to acquired cells (GV: 63.6 +/- 8.4%, PLV: 60.5+/-5.4%, P=0.73). However, the proportion of macrophages was significantly increased (GV: 5.6 +/-1.5, PLV: 14.0+/-1.3, P=0.006). These results suggest that the anti inflammatory effect of PLV is not caused by the change in CD11b expression, and that PLV affects the proportion of macrophage among BALF cells. PMID- 12908012 TI - Attempt to establish an experimental animal model of moyamoya disease using immuno-embolic material--histological changes of the arterial wall resulting from immunological reaction in cats. AB - In this study, we investigated the relationship between intimal thickening of the internal carotid artery (ICA) and immunological reaction, and between occlusion of the ICA and development of basal collateral vessels in moyamoya disease. Rod shaped lactic acid-glycolic acid copolymer (LGA-50) and N-acetylmuramyl-L-alanyl D-isoglutamine (muramyl dipeptide: MDP), and immuno-embolic material, were injected into cats unilaterally via the common carotid artery. Histological changes of duplication of the internal elastic lamina could be seen mainly in the terminal portion of the ICA in the animals injected with rod-shaped LGA-50 containing MDP. No angiographic changes were seen in any of the animals. These findings suggest that the immunological reaction induced by MDP caused histological changes in the intima of the ICA similar to those observed in moyamoya disease. This experimental study, however, could not clarify the development of the basal collateral vessels. PMID- 12908013 TI - Efficacy of interferon retreatment on interferon-resistant patients with chronic hepatitis C. AB - Chronic Hepatitis C can progress to end-stage liver cirrhosis or hepatocellular carcinoma. Interferon (IFN) therapy is effective in clearing the hepatitis C virus and in improving liver histology, however, few patients maintain a sustained response (SR) after IFN withdrawal. Immediate retreatment with IFN is therefore considered to be both effective and necessary, especially for patients who do not respond to the initial course of IFN therapy. All 145 patients included in the present study underwent liver biopsy, followed by a first treatment course with various IFNs (alpha2a, alpha2b, alpha, OIF or beta). If hepatitis C virus (HCV) RNA was positive after the first treatment, the patient was assigned to one of 3 groups, depending on whether his or her alanine transaminase (ALT)level was normalized (incomplete response, IR), partially responsive(PR), or non-responsive (NR). After an observational interval of 6 to 76 months, a second IFN treatment was initiated with a higher dose or the same dose of the same IFN for the IR group, and with a different IFN for the PR and NR groups. At 6 months after retreatment with IFN, the overall efficacy of the retreatment was 29.7.% In the case of the IR group, who received the same IFN, the overall efficacy was 45.2%. In patients identified as non-SR after the first treatment, who received a different type of IFN for retreatment, the overall efficacy was 18.6%. Anti-IFN antibody was not detected in most of the breakthrough cases. For some IR patients, retreatment with the same IFN was effective. Anti-IFN antibody was mostly negative, indicating that the same IFN can be used in both the first treatment and retreatment to obtain an SR. Switching to a different IFN was effective for some PR and NR patients, suggesting that changing IFN for such cases is a good therapeutic choice. PMID- 12908014 TI - Absence of scalenus anterior muscle. AB - A rare anomaly of the scalenus muscles is described. In this case, the right scalenus anterior muscle was absent. As a substitute for this muscle, some aberrant muscle slips arose from the lower vertebrae and descended in front of the ventral rami of the lower cervical nerves. These aberrant slips then ran between the ventral rami of the the eighth cervical and first thoracic nerves, and were fused with the right scalenus medius muscle. Thus, the subclavian artery and vein ran in front of the aberrant slips, together with the ventral ramus of the first thoracic nerve. The aberrant muscle slips issued 2 accessory bundles. One bundle ran between the ventral rami of the fourth and fifth cervical nerves and was fused with the scalenus medius muscle; the other bundle ran between the ventral rami of the fifth and sixth cervical nerves and was fused with the scalenus medius muscle. PMID- 12908015 TI - "The state has no business in the bedrooms of the nation". PMID- 12908016 TI - Clinical use of sonohysterography in the evaluation of infertility. AB - Assessment of tubal patency and evaluation of the uterine cavity should be part of all infertility investigations. Sonohysterography (SHSG), also referred to as saline-enhanced ultrasonography, is being used with increasing frequency for investigation of infertility and other gynaecological indications. Sonohysterography is similar to hysterosalpingography (HSG) in its effectiveness for evaluating tubal patency, and superior to HSG for assessing intrauterine abnormalities. Advantages of SHSG over HSG include the ability to assess extrauterine structures, lack of ionizing radiation, and often better tolerability by the woman being examined. Sonohysterography can offer detailed assessment of the female pelvis, and has the potential to replace HSG as a routine, first-line infertility investigation. This paper discusses the technique and indications for SHSG, and reviews its use in infertility investigation, in particular in the assessment of the uterine cavity and tubal patency. PMID- 12908017 TI - The maternal benefits of corticosteroids with HELLP (hemolysis, elevated liver enzymes, low platelet count) syndrome. AB - OBJECTIVES: To determine the more effective regime in improving hematologic abnormalities associated with HELLP (hemolysis, elevated liver enzymes, low platelet count) syndrome, by comparing two different antenatal dosing protocols for dexamethasone (given primarily for fetal lung maturity). STUDY DESIGN: A retrospective cohort study of 30 women between 24 and 34 weeks' gestation, diagnosed with HELLP syndrome prior to delivery, who received 2 doses of dexa methasone 12 mg intramuscularly, given either 24 hours apart or 12 hours apart. RESULTS: There was a shorter time from the first corticosteroid dose to the beginning of improvement with the 12-hour regime for platelet count (11 hours [9, 25 hours] versus 69 hours [41, 112 hours], median [quartiles], P = 0.003) and for alkaline phosphatase (25 hours [5, 44 hours] versus 59 hours [31, 69 hours], median [quartiles], P = 0.02). Worsening recurred from 11 hours, for alanine aminotransferase (ALT) and aspartate transaminase (AST), to 32 hours, for uric acid, after initial improvement. Hematologic improvement occurred in 37% (for ALT) to 67% (for alkaline phosphatase) of women overall. CONCLUSION: A temporary improvement in the hematologic parameters of HELLP syndrome can occur with antenatal dexa-methasone administration, with the 12-hour regime having quicker onset to improvement in platelet count and alkaline phosphatase. PMID- 12908018 TI - Do maternal characteristics and past pregnancy experiences predict preterm delivery among women aged 20 to 34? AB - OBJECTIVE: To identify key demographic, lifestyle, and medical indicators for preterm births in women aged 20 to 34. METHODS: A population-based, case-control study consisting of 987 women aged 20 to 34, who delivered a liveborn singleton infant at <37 weeks' gestation in Alberta between May 1999 and August 2000. Controls delivered liveborn infant(s) at > or =37 weeks' gestation. Information was obtained from computer-assisted telephone interviews, and the provincial Physician Notice of Stillbirth/Birth database. Analysis included bivariate and logistic regression techniques. RESULTS: Logistic regression modelling indicated that significant risk factors for preterm delivery included poor past pregnancy outcome (odds ratio [OR] 6.4), poor emotional health (OR 1.8), more than 3 years or less than 1 year between pregnancies (OR 1.4 and 1.9, respectively), polyhydramnios and oligohydramnios (OR 4.1), bleeding at greater than 20 weeks' gestation (OR 10.4), malpresentation (OR 2.9), gestational hyper- tension (OR 2.2), and gestational hypertension with proteinuria (OR 4.4). Women who had fewer than 10 prenatal visits, regardless of attending prenatal classes, were at highest risk of preterm delivery (OR 6.7). CONCLUSIONS: In this population of women aged 20 to 34 years, few prenatal visits, poor emotional health prior to pregnancy, and conditions of the current pregnancy were strongly associated with preterm singleton birth. PMID- 12908019 TI - Arrest of descent in second stage of labour secondary to macrosomia: a case report. AB - BACKGROUND: Fetal macrosomia, defined as birth weight greater than 4000 g, complicates 10% of pregnancies and is a well-documented cause of prolonged second stage of labour, as well as of arrest of descent of the fetal presenting part. CASE: A multigravida woman with gestational diabetes mellitus was admitted in labour at term, and progressed to full dilatation. The fetal vertex failed to descend beyond -3 station. An emergency Caesarean section was performed and a 6452 g male infant was delivered. CONCLUSION: Physicians should be aware of the possibility of macrosomia as the cause of failure of descent in the second stage. A heightened state of suspicion should be maintained, particularly in a multigravida woman with a prior macrosomic baby and the presence of other predisposing factors such as gestational diabetes mellitus. PMID- 12908020 TI - Emergency contraception. AB - OBJECTIVE: To review current knowledge about emergency contraception (EC), including available options, their modes of action, efficacy, safety, and the effective provision of EC within a practice setting. OPTIONS: The combined estradiol-levonorgestrel (Yuzpe regimen) and the levonorgestrel-only regimen, as well as post-coital copper intrauterine devices, are reviewed. OUTCOMES: Efficacy in terms of reduction in risk of pregnancy, safety, and side effects of methods for EC and the effect of the means of access to EC on its appropriate use and the use of consistent contraception. EVIDENCE: MEDLINE and the Cochrane Database were searched for English-language articles published from January 1998 through March 2003, to update the previous SOGC guidelines published in 2000. Clinical guidelines and position papers developed by health or family planning organizations were also reviewed. Key words used were: emergency contraception, post-coital contraception, emergency contraceptive pills, postcoital copper IUD. VALUES: The studies reviewed were classified according to criteria described by the Canadian Task Force on the Periodic Health Exam and the recommendations for practice were ranked based on this classification. BENEFITS, HARMS, AND COSTS: These guidelines are intended to help reduce unintended pregnancies by increasing awareness and appropriate use of EC. RECOMMENDATIONS: 1. Women who have had unprotected intercourse and wish to prevent pregnancy should be offered hormonal EC up to 5 days after intercourse. (II-2A) 2. A copper IUD can be used up to 7 days after intercourse in women who have no contraindications. (III-B) 3. Women should be advised that the levonorgestrel EC regimen is more effective and causes fewer side effects than the Yuzpe regimen. (I-A) 4. Either 1 double dose of the levonorgestrel EC regimen (1.5 mg) or the regular 2-dose levonorgestrel regimen (0.75 mg each dose) may be used, as they have similar efficacy with no difference in side effects. (I-A) 5. Hormonal EC should be started as soon as possible after unprotected sexual intercourse. (II-2B)6. Women of reproductive age should be provided with a prescription for hormonal EC in advance of need. (I-A) 7. The woman should be evaluated for pregnancy if menses have not begun within 21 days following EC treatment. (III-A) 8. A pelvic examination is not indicated for the provision of hormonal EC. (III-A) Validation: These guidelines have been reviewed by the Clinical Practice Gynaecology and Social and Sexual Issues Committees of the Society of Obstetricians and Gynaecologists of Canada. SPONSOR: The Society of Obstetricians and Gynaecologists of Canada. PMID- 12908022 TI - A dip into history: science and publication in cryobiology 150 years ago. PMID- 12908021 TI - Tension-free vaginal tape (TVT) procedure. AB - OBJECTIVE: To provide an introduction to the tension-free vaginal tape (TVT) procedure, placing it in the context of minimally invasive surgeries for the treatment of urinary stress incontinence in women, and to provide guidance to surgeons counselling women on the merits of this surgical choice. OPTIONS: This discussion is limited to surgical treatment of urinary stress incontinence in women. EVIDENCE: A search of both MEDLINE and the Cochrane Library identified the most relevant medical evidence. This document represents an abstraction of the evidence rather than a methodological review. VALUES: This update is the consensus of the Sub-Committee on Urogynaecology of the Society of Obstetricians and Gynaecologists of Canada (SOGC). BENEFITS, HARMS, AND COSTS: Counselling for the surgical treatment of urinary incontinence should consider the harms-benefits ratios of various options and include a measure of the "weight" of the evidence. RECOMMENDATIONS: 1. The Burch procedure should be offered as the gold standard. The TVT procedure is promising but currently under evaluation in trials that will establish its efficacy and safety (II-3A). 2. Proper training is recommended prior to performing TVT procedures. 3. Long-term trial results are needed before the TVT procedure can be offered to patients as an equal alternative to the Burch procedure. VALIDATION: This technical update has been approved by the Sub Committee on Urogynaecology and Executive and Council of the SOGC. PMID- 12908023 TI - Novel thermal hysteresis proteins from low temperature basidiomycete, Coprinus psychromorbidus. AB - The low temperature basidiomycete, Coprinus psychromorbidus, produced three kinds of thermal hysteresis proteins (THPs) in the extracellular space. Molecular mass of purified fungal THPs were approximately 23 kDa, respectively, however those fungal THPs had different N-terminal amino acid sequences. Those fungal THPs formed ice crystals resembling "stone Age arrow head". These observations indicate that fungal THPs did not form hexagonal ice crystals to inhibit its growth. PMID- 12908024 TI - A simplified measurement of hydrated crystal densities of low melting points (low transition points) by solidifications of aqueous solutions. AB - A volume change method for measuring crystal densities is described. It allows the densities of unstable hydrated crystals at room temperature to be determined, by measurements of volume changes during the solidification of aqueous solutions. NaCl x 2H2O, KCl, MgSO4 x 12H2O and K2HPO4 x 6H2O were measured by the method and their densities (SE) are 1.61+/-0.02, 1.99+/-0.05, 1.45+/-0.01 and 1.75+/-0.02 g ml(-1) respectively. Data of NaCl x 2H2O and KCl are in good agreement with the previously reported values. PMID- 12908025 TI - Development of probabilistic tools to assist in the establishment and management of cryopreserved plant germplasm collections. AB - A simple method, based on the binomial distribution, is proposed to calculate the probability of recovering at least one (or any other fixed number of) plant(s) from a cryobank sample using four given parameters: the percentage of plant recovery observed from a control sample, pobs, the number of propagules used for this control, n1, the number of propagules in the cryobank sample, n2, a chosen risk for the calculation of a confidence interval for the observed plant recovery, alpha. Using this method, it is possible to assess the number of propagules which should be rewarmed immediately after freezing in order to estimate the plant recovery percentage as a function of the total number of propagules available. It also allows the calculation of the minimum plant recovery percentage to ensure that the probability to recover at least one (or A, with A>1) plant(s) is higher than a fixed probability level, as a function of the control and the cryobank sample sizes. Reciprocally, once the plant recovery percentage has been estimated, it is possible to assess the minimum size of the cryobank sample to obtain a probability to recover at least one (or A, with A>1) plant(s) higher than some fixed level. PMID- 12908026 TI - A numerical study of cell behaviour in a ternary solution during the freezing process. AB - Using a continuum model for multi-component phase change system, the freezing of cell suspension in a ternary solution, H2O-NaCl-CPA (cryoprotective agent) inside a flat bag is investigated numerically in this study. The temperature and phase change history, intracellular water loss, and the volume change of the cells at different locations inside cell suspension are calculated. Numerical results reveal that although the sample boundary is cooled at a constant rate, different locations inside the sample experienced different temperature changes and cooling rates. The highest cooling rates occur at internal locations. The cell volume change is location-dependent. PMID- 12908027 TI - Evaluation of damage in Pacific oyster (Crassostrea gigas) spermatozoa before and after cryopreservation using comet assay. AB - We examined the applicability of the comet assay (single cell gel electrophoresis assay) to estimate the quality of frozen-thawed Pacific oyster (Crassostrea gigas) spermatozoa. Comet assay was performed on semen before and after cryopreservation followed by fluorescent staining with propidium iodide to assess DNA integrity. After cryopreservation, the percentage of spermatozoa with damaged DNA significantly increased, while only about half of the cells displayed intact DNA, even when protected with 10 percent DMSO. All the considered parameters (head length, head area, head intensity, total length, total area, total intensity, tail length percent, tail area percent, and tail intensity percent) were higher than the oyster sperm protected with 10 percent DMSO-artificial sea water after freezing and thawing. Only tail length percent, tail area percent, and tail intensity percent were increased significantly after cryopreservation. The tail length percent was found to be the most sensitive indicator of the cryopreservation-induced DNA damage. Our freeze-thawing procedure significantly affected oyster sperm DNA, as indicated by the reduced fertilization rate when frozen-thawed oyster sperm are used. Irreversible alteration of the genome may prevent fertilization or alter normal embryonic development. This study is the first to demonstrate that the comet assay is an inexpensive, rapid and sensitive method for determining DNA damage in Pacific oyster sperm quality assessments. PMID- 12908028 TI - Cryopreservation of shoot tips of Picrorhiza kurroa Royle ex Benth, an indigenous endangered medicinal plant, through vitrification. AB - The cryopreservation of shoot tips of Picrorhiza kurroa Royle ex Benth (IC 266698), an endangered medicinal plant of India was investigated. Shoot tips (about 1 mm in length) excised from four-week-old proliferating shoot cultures were precultured on MS medium supplemented with various osmotica before dehydrating with PVS2 solution at 0 degrees C. The dehydrated shoot tips were directly immersed in LN2. Following cryopreservation, and after rapid rewarming at 45 degrees C, shoot tips were quickly washed with 1.2 M sucrose solution and then plated on solidified shoot culture medium. Shoot tips were successfully cryopreserved by vitrification, when they were precultured on medium supplemented with 5% DMSO at 4 degrees C for two days before dehydrating in PVS2 for 10-20 minutes at 0 degrees C. Average survival in terms of normal shoot formation after 4 wks of plating was about 20% without callus formation. Cold hardening of shoot cultures for four weeks at 4 degrees C significantly improved the survival and shoot regeneration of cryopreserved shoot tips to 70% and 35%, respectively. PMID- 12908029 TI - Selection of overwintering microhabitats used by the arctic woollybear caterpillar, Gynaephora groenlandica. AB - In extreme environments such as the High Arctic, climatic conditions challenge physiological tolerance of insects resulting in prolonged dormancy and extended life cycles. Therefore, the selection of suitable microhabitats for overwintering is crucial. At two field sites on Ellesmere Island, we located hibernacula (silk overwintering structures) used by the Arctic woollybear caterpillar, Gynaephora groenlandica (Wocke) (Lepidoptera: Lymantriidae) during their 11 month dormancy. All hibernacula found were anchored to the base of rocks and were not associated with vegetation. Rocks may function to absorb solar radiation and re-radiate heat, accelerating localized snowmelt, thus allowing caterpillars to emerge as early as possible in spring to begin foraging. Temperatures experienced by G. groenlandica in hibernacula throughout the winter were well within their physiological tolerance range. During late summer aestivation, hibernaculum temperatures were similar to, but more stable than, nearby soil surface temperatures. Lower maximum daily hibernaculum temperatures during the warmest month (July) may reduce metabolic rates and provide some energy savings. Since hibernacula were not randomly distributed around rocks, this suggests that G. groenlandica key in on some feature of the hibernaculum site. The northeast orientation of hibernacula at Eastwind Lake corresponds to the leeward side of rocks. Therefore wind patterns may be important in hibernaculum site selection. Other potential cues that may guide selection of hibernacula sites remain unclear: no seasonal changes in preference for light or soil moisture were observed between active and dormant caterpillars, and thermotaxis could not be distinguished from thigmotaxis. PMID- 12908030 TI - [Prevalence of varicella-zoster virus antibodies in young adults from different Brazilian climatic regions]. AB - To evaluate the prevalence of varicella-zoster virus infection in young adults from different Brazilian urban regions, 975 serum samples from blood donors aged 20 to 29 years, from tropical climate cities (Salvador and Fortaleza) and from temperate climate cities (S o Paulo, Curitiba and Porto Alegre) were tested by an in-house ELISA for detection of anti-varicella-zoster virus IgG antibodies. The overall prevalence was 94.2%. The lowest rate was observed in Fortaleza (88.7%) and the highest in Curitiba (99.5%). Seroprevalence in tropical regions of Brazil (89.4%) was significantly higher than in temperate regions (97.3%), a similar pattern to that observed in other tropical countries. PMID- 12908031 TI - Feasibility study of the immunogenicity and safety of a novel DTPw/Hib (PRP-T) Brazilian combination compared to a licensed vaccine in healthy children at 2, 4, and 6 months of age. AB - Vaccination of infants with conjugated Haemophilus influenzae type b (Hib) vaccines has been proven to reduce Hib meningitis by 95% and pneumoniae by 20%. The routine use of Hib vaccine is facilitated by the introduction of combination vaccines into the EPI (Expanded Plan of Immunization). The objective of this study was to compare the immunogenicity and reactogenicity of an extemporaneously mixed DTPw/Hib (diphtheria-tetanus-whole cell pertussis) combination, using the technology of two Brazilian manufacturers, against a licensed DTPw/Hib European combination in 108 infants vaccinated at 2, 4 and 6 months according to the local national schedule. The Brazilian combination was highly immunogenic with Hib seroprotection rates (anti-PRP > 0.15 mg /ml of 98% after 2 doses and 100% after 3). Also for tetanus and pertussis the new Brazilian combination was as immunogenic as the European counterpart, except the diphtheria seroprotection rates and titers were lower. There was also no clinically relevant difference in reactogenicity. If these feasibility results are confirmed, the Brazilian DTPw/Hib combination should help to boost the uptake of Hib vaccination in Brazil. PMID- 12908032 TI - [Parasitological study with faecal samples of stray and domiciliated cats (Felis catus domesticus) from the Metropolitan Area of Rio de Janeiro, Brazil]. AB - In a survey of 131 faecal samples from Rio de Janeiro domiciliated and stray cats, 63,4% were positive with a higher prevalence among stray animals. Ancylostoma sp (43.5%), Toxocara sp (19.1%), Cystoisospora sp (43.5%), Uncinaria sp (1.5%), Toxocaris leonine (7.6%), Giardia sp (6.1%) and Sarcocystis sp (0.8%) were found. This high prevalence suggests the need for improved control measures against cats to protect animal and human health. PMID- 12908033 TI - Chemotherapeutic effects on larval stages of Schistosoma mansoni during infection and re-infection of mice. AB - The sensitivity of the larval stages of Schistosoma mansoni to chemotherapy with praziquantel and oxamniquine was tested in mice during primary and secondary infections and after different intervals from cercarial exposure. Worm recovery by perfusion of the porto-mesenteric system, followed by counting and a morphometric study of the parasite, allowed the conclusion that the relative resistance of the larval stages of S. mansoni to schistosomicide drugs, demonstrated in primary infections, also persists when the host is already infected. This indicates that a therapeutic failure may result when an infected host is treated some time after being re-infected, because of the presence of migrating, drug-resistant, immature forms of the parasite. PMID- 12908034 TI - [Culicidae insect fauna from rural zone in Amazonas State with incidence of sylvatic yellow fever]. AB - After the occurrence of 14 sylvatic yellow fever cases in 10 cities in the State of Amazonas during 1996, an investigation into the presence of sylvatic yellow fever vectors was carried out. The material of larvae and adult insects was collected around residences and canopy trees within forests, using a light trap (CDC) and human bait. A total of 424 insects was collected. Thirty seven species were identified, some of which were sylvatic yellow fever vectors: Haemagogus janthinomys, Ha. leucocelaenus, Aedes fulvus. PMID- 12908035 TI - [Epidemiological and clinical aspects of scorpion envenomation in the region of Santarem, Para, Brazil]. AB - This is a descriptive and prospective study on epidemiological and clinical aspects of 72 scorpion accidents admitted to Santarem Municipal Hospital, state of Para, Brazil, from February 2000 to February 2001. Only 8.3% brought the animal with them, identified as T. cambridgei. The majority of victims were male (83.3%). The mean age and the time of the medical help were respectively 33.6 +/- 18.3 years and 4.6 +/- 3.2 hours. The parts of the body most affected were the superior members (51.5%). Local symptoms occurred in 91.7% cases and systemic manifestations in 98.6% of the accidents. The local symptoms included: paresthesia in 79.2% cases, pain in 52.8% and edema in 26.4%. Among the systemic manifestations neurological disorders predominated in 97.2%, and the symptom of "electric shock" occurred in 88.9% patients. The most common neurological signs were: myoclonia (93%), dysmetria (86.1%), dysarthria (80.6%), and ataxia (70.8%). The accidents were classified as moderate in 76.4% without any serious cases. The specific anti-venom serum was not administered in 32.7% of the moderate cases, due to non-availability of the anti-venom serum at the time of attendance. The victims of scorpion envenomation notified at Santarem, present a different clinical and regional behavior from previous reports in Brazil and Amazonia regions. The predominantly neurological picture has not previously been described in the Brazilian literature. PMID- 12908036 TI - [Antibodies to the rabies virus in humans working in a veterinary hospital environment at the Universidade Federal de Mato Grosso. Cuiaba, MT, Brazil]. AB - A characterization of the serological profile of 102 workers in constant activities in a veterinary hospital environment at the Universidade Federal de Mato Grosso was made through epidemiological data and serological tests to the rabies virus from November 1999 to November 2000. Of these, 27 had been vaccinated in a pre-exposure scheme and 75 had not received any kind of antirabies immunization. The results of the study classified the people into four groups (G1, G2, G3 and G4). In G1, 19 (18.6%) people that had been previously vaccinated showed a titration level lower than 0.5UI/mL; in G2, 8 (7.8%) people who had been previously vaccinated showed a titration level higher than 0.5UI/mL; in G3, 67 (65.6%) people who had not been vaccinated against rabies virus showed a titration level lower than 0.5UI/mL; and finally in G4, 8 (7.8%) people who had never received any kind of vaccine scheme showed a titration level higher than 0.5UI/mL. The results showed that epidemiological evaluation and serological profile are necessary people working in a veterinary hospital environment while being pre-exposure vaccinated. PMID- 12908037 TI - [Vectorial transmission of Chagas' disease in Mulungu do Morro, Northeastern of Brazil]. AB - A serological survey was carried out to determine the prevalence of Chagas' disease in municipality. The following variables were analyzed to identify the form of transmission of this disease: age, sex, clinical and transfusional history, degree of kinship and serology. Within the 863 municipalities we studied, we identified 265 individuals, with serology testing done on them and on their respective mothers. Of these, 232 tested negative serology for Chagas'disease and 33 (14.2%) positive. We found 9 (3.9%) patients, of 14.3 years. average age with vectorial transmission and 24 (10.3%), of 26.6 years. average age with probable, vertical and vectorial transmission. When we compare the two groups in regard to age averages and manner of transmission of Trypanosoma cruzi, we encounter a statistical significance. Our results suggest the existence of an active, vectorial transmission of Trypanosoma cruzi in Mulungu do Morro. PMID- 12908038 TI - [A comparative study between the efficacy of pentamidine isothionate given in three doses for one week and N-methil-glucamine in a dose of 20mgSbV/day for 20 days to treat cutaneous leishmaniasis]. AB - Seventy-nine patients with cutaneous leishmaniasis were included in this study. The experimental group (n = 38) was treated with pentamidine isothionate in a dose of 4mg/kg/day on alternate days, for one week. The control group (n = 41) was treated with N-methylglucamine in a dose of 20mgSbV/kg/day for 20 days. Twenty-one isolates were identified using monoclonal antibody technique. We characterized Leishmania (Viannia) braziliensis, most frequently. There was a cure rate of 71.05% of the patients in the experimental group and 73.17% in the control group (p = 0.47). We found a statistical significance regarding frequency of ECG alterations between the experimental and control group (p<0.05). In our study pentamidine was as effective as antimonial for the treatment of american cutaneous leishmaniasis. It proved to be a safer drug considering heart toxicity. Moreover, it requires less time to complete the treatment. PMID- 12908039 TI - [Leprosy in Brazil]. AB - Leprosy or Hansen's disease is a chronic infectious disease caused by the Mycobacterium leprae. The skin and nervous manifestations of the disease present a singular clinical picture that is easily recognized. After India, Brazil still is the second country with the greatest number of cases in the world. Around 94% of the known cases and 94% of the new cases reported in America, come from Brazil. The disease presents itself in two well-defined stable and opposite poles (lepromatous and tuberculoid) and two unstable groups (indeterminate and dimorphic). The spectrum of presentation of the disease may also be classified as: tuberculoid tuberculoid (TT), borderline tuberculoid (BT), borderline borderline (BB), borderline lepromatous (BL) and lepromatous lepromatous (LL). The finding of acid fast bacillus in tissue is the most useful method of diagnosis. The effective treatment of leprosy includes the use of specific therapy, suppression of lepra reactions, prevention of physical incapacity, and physical and psychosocial rehabilitation. Chemotherapy with rifampin, dapsone and clofazimine have produced very good results and the control of the disease in Brazil in the foreseeable future is likely. PMID- 12908040 TI - [Internal validity of clinical trials for Plasmodium vivax malaria treatment: analysis of evaluation study of in vivo Plasmodium vivax emergence of resistance to standard doses of primaquine]. AB - Parasite resistance can be defined as the ability of a parasite strain to survive and/or to multiply despite the administration and absorption of a drug given in doses equal or higher than those usually recommended, but within the limits of tolerance of the patients. Therefore, the ideal study design to be used to monitor emergence of parasite resistance would use historical controls or any valid baseline data. Moreover, it is desirable to have some control of remaining determinants of therapeutic failure, not related to biological parasite resistance, which could vary over time and would have potential to distort the analysis for detecting emergence of parasite resistance. Here we comment on the internal validity of studies, which aim to assess the in vivo Plasmodium vivax emergence of resistance to standard doses of primaquine used routinely by health services. Few studies have paid attention to the need to control for other determinant factors of therapeutic failures or made any attempt to compare current findings with cure failure rates of historical controls from a given geographical area. Therefore, attention to the internal validity (and limitations) of study conclusions in these types of study is strongly advised. PMID- 12908041 TI - [Hepatitis A]. AB - Hepatitis A infection is known since the ancient Chinese, Greek and Roman civilizations but the first documented report was published in the eighteenth century. The hepatovirus belongs to the Picornaviridae family, and carries a single strand RNA. There are 7 genotypes. Antibodies of the IgM and IgA classes, during natural infections, appear early in the serum, together with the first clinical manifestations of the disease, but they may also appear at the end of the first week of infection. There is a spectrum of clinical presentation: asymptomatic infection, symptomatic without jaundice and symptomatic jaundiced. A rare fatal form of hepatitis has been described. Diagnosis of the hepatitis A infection is confirmed by the finding of IgM anti-HAV antibodies, routinely performed using an ELISA test. Treatment is supportive. Intramuscular anti-A gamma globulin is used for passive immune prophylaxis, and there is an efficient vaccine for active immune prophylaxis. PMID- 12908042 TI - [Dilated cardiomyopathy in patients with chronic chagasic infection: report of two fatal autochthonous cases from Rio Negro, State of Amazonas, Brazil]. AB - Two fatal cases of human chronic chagasic cardiomyopathy are reported, for the first time, in autochthonous patients from Rio Negro, Amazonas state. Both cases, (45 year old man and 44 year old woman) who were born and lived their whole lives in the Rio Negro region, in the northern part of the state of Amazonas, reported having been bitten several times by triatomine bugs in their camping huts while gathering pia ava fibers. The patients, who had confirmed positive serology for Trypanosoma cruzi antibodies (indirect immunofluorescence, ELISA and Western blot), developed in the last 5-7 years a progressive cardiac failure, with global enlargement of the heart, atrioventricular and left bundle branch block, ventricular extrasystoles, both dying from irreversible cardiac insufficiency. The histopathology of a post-mortem surgical cardiac biopsy performed in one of the cases showed chronic myocarditis with diffuse mononuclear cell infiltrates, with areas of focal cell accumulation, fibrosis, lytic necrosis of myocardial fibers, suggestive of chronic chagasic myocarditis. An in situ PCR was positive for Trypanosoma cruzi. PMID- 12908043 TI - [Extraintestinal finding of Isospora belli unizoic cysts in a patient with AIDS: case report]. AB - This report describes the presence of Isospora belli unizoic cysts in mesenteric lymph nodes and of gametocytes in the gallbladder epitelium of a 26 year-old Brazilian male patient with Acquired Immune Deficiency Syndrome. This patient had received treatment for several times with sulfamethoxazole-trimethoprim. It is discussed the significance of I. belli tissue cysts as possible foci of resistance of the parasite and their association with the infection relapse even post-treatment with anticoccidian medication. PMID- 12908044 TI - [Immediate and generalized reaction to Montenegro skin test]. AB - The authors describe for the first time a case of immediate and generalized reaction to Montenegro skin test with merthiolated antigen. This reaction consisted of generalized cutaneous rash with pruritus, and was treated with oral bromethazine. The pruritus disappeared an hour after medication and the rash 12 hours later. PMID- 12908045 TI - [Blastocystosis: nitazoxanide as a new therapeutic option]. AB - We report a case of a patient with AIDS and severe immunosuppression who developed diarrhea due to a vacuolar stage of Blastocystis hominis. The patient was treated with nitazoxanide, a 5-nitrothiazole broad-spectrum antiparasitic agent. We obtained complete remission of symptoms and eradication of the parasite. Nitazoxanide is a new treatment option for blastocystosis in AIDS patients. Further studies to test its efficacy are needed. PMID- 12908046 TI - [Epidemiologic investigation into characteristics of canine and feline population in a district close to a rural area in Cuiaba-MT, with a view to control of animal rabies]. AB - Questionnaires answered by 476 students (age 15-20 yrs) attending a local school in the neighborhood of Pedra 90 in Cuiaba -MT were analyzed in order to study the characteristics of feline and canine populations in the region. The results showed that 371 (78%) of the 476 families kept 513 dogs and 307 cats. Of 573 dogs, 289 (56.3%) were male and 224 (43.7%) were female. Of 307 cats, 182 (59.28%) were male and 125 (40.72%) were female. The owners of 474 (92.4%) dogs and 267 (86.9%) cats reported that their pets had been vaccinated against rabies. PMID- 12908047 TI - [Occurrence of Cryptosporidium sp in fecal samples of children less than 10 years old with clinical indication of Rotavirus]. AB - The presence of oocysts of Cryptosporidium sp was investigated in 485 fecal samples of children with clinical indication of Rotavirus. No significant differences were observed between Cryptosporidium sp. and rotavirus occurrence and fecal consistency. Cryptosporidium sp also should be performed in the laboratory diagnosis of diarrheic episodes in children. PMID- 12908048 TI - First record on Loxosceles laeta (Nicolet, 1849) (Araneae, Sicariidae) in the West Zone of Sao Paulo City, Sao Paulo, Brazil, and considerations regarding its geographic distribution. AB - Loxosceles laeta spiders were captured in the West zone of Sao Paulo City, this being the first record of the specie in this area. Since loxoscelism is an important public health problem in the South region of Brazil, it is necessary to investigate the presence of this spider in Sao Paulo City. PMID- 12908049 TI - [Evaluation of cytopathologic exam for diagnosis of oral chronic paracoccidioidomycosis]. AB - With the aim of evaluating exfoliative cytology for the diagnosis of paracoccidioidomycosis oral lesions, eight patients that presented the disease were studied. The presence of fungi was demonstrated in all these cases. It was concluded that the oral exfoliative cytology exam can be effectively used in the diagnosis of paracoccidioidomycosis and contribute to the therapeutic control of oral forms of this mycosis. PMID- 12908050 TI - [Bacterial colonization in a hemodialysis dual lumen temporary catheter]. PMID- 12908052 TI - Neuroendocrine alterations impair enamel mineralization, tooth eruption and saliva in rats. AB - Neonatal administration of monosodium glutamate (MSG) in rats causes definite neuroendocrine disturbances which lead to alterations in many organ systems. The possibility that MSG could affect tooth and salivary gland physiology was examined in this paper. Male and female pups were injected subcutaneously with MSG (4 mg/g BW) once a day at the 2nd, 4th, 6th, 8th and 10th day after birth. Control animals were injected with saline, following the same schedule. Lower incisor eruption was determined between the 4th and the 10th postnatal days, and the eruption rate was measured between the 43rd and the 67th days of age. Pilocarpine-stimulated salivary flow was measured at 3 months of age; protein and amylase contents were thereby determined. The animals treated with MSG showed significant reductions in the salivary flow (males, -27%; females, -40%) and in the weight of submandibular glands (about -12%). Body weight reduction was only about 7% for males, and did not vary in females. Saliva of MSG-treated rats had increased concentrations of total proteins and amylase activity. The eruption of lower incisors occurred earlier in MSG-treated rats than in the control group, but on the other hand the eruption rate was significantly slowed down. The incisor microhardness was found to be lower than that of control rats. Our results show that neonatal MSG treatment causes well-defined oral disturbances in adulthood in rats, including salivary flow reduction, which coexisted with unaltered protein synthesis, and disturbances of dental mineralization and eruption. These data support the view that some MSG-sensitive hypothalamic nuclei have an important modulatory effect on the factors which determine caries susceptibility. PMID- 12908053 TI - Oral health profile of education and health professionals attending handicapped children. AB - The purpose of this study was to evaluate the knowledge and attitudes toward oral health of education and health professionals working in a children care program for handicapped children from 0 to 6 years of age, run by a public municipal institution in Rio de Janeiro. Using a printed questionnaire, 67 professionals (teachers, attendants and health professionals) were interviewed. The results were compared to the children's oral hygiene habits, by directly observing their daily nursery routine. Although 97.0% said that oral health could play a part in general health, only 37.3% of the professionals answered correctly on this matter. As for methods for preventing caries, although 92.5% said that they were aware of them, only 17.9% went to the dentist for preventive treatment. Although the majority (81.3%) indicated oral hygiene as a way of preventing caries, observation showed that this practice is not always put into effect in the program's day nursery. Regarding when to start toothbrushing in children, 75.0% of the teachers and 94.4% of the health professionals said that they were aware of the need to begin brushing before one year of age, although this reply was given by only 52.5% of the attendants (chi-square, p = 0.006). In view of these results, it was concluded that attitudes toward oral health were not always coherent with the knowledge that these professionals express. PMID- 12908054 TI - Effectiveness of low cost toothbrushes, with or without dentifrice, in the removal of bacterial plaque in deciduous teeth. AB - The main objective of this study was to compare the effectiveness of a low cost toothbrush ("monoblock") to that of a conventional toothbrush with and without addition of dentifrice with respect to the removal of dental plaque. Thirty-two 4 to 6-year-old children took part in this study: they were evaluated under four experimental conditions defined by the combinations of the values of two factors, toothbrush (conventional or monoblock) and use of dentifrice (with or without). The effectiveness of the treatments was defined in terms of the reduction of a bacterial plaque index evaluated before and after toothbrushing. No statistically significant differences were detected between the two types of toothbrushes with respect to the reduction of the bacterial plaque index. Similarly, there were no statistical evidences that the use of dentifrice improves the mechanical control of dental plaque. These results are important from a public health point of view, specially in developing countries, where the dissemination of educational and preventive techniques of low cost are fundamental. PMID- 12908055 TI - Applicability of 3D-CT facial reconstruction for forensic individual identification. AB - Computed tomography (CT) is used in several clinical dentistry applications even by axial slices and two and three-dimensional reconstructed images (2D-CT and 3D CT). The purpose of the current study is to assess the precision of linear measurements made in 3D-CT using craniometric patterns for individual identification in Forensic Dentistry. Five cadaver heads were submitted to a spiral computed tomography using axial slices, and 3D-CT reconstructions were obtained by volume rendering technique with computer graphics tools. Ten (10) craniometric measurements were determined in 3D-CT images by two examiners independently, twice each, and the standard error of intra- and inter-examiner measurements was assessed. The results demonstrated a low standard error of those measurements, from 0.85% to 3.09%. In conclusion, the linear measurements obtained in osseous and soft tissue structures were considered to be precise in 3D-CT with high imaging quality and resolution. PMID- 12908056 TI - [Normative values and sexual dimorphism in aesthetically pleasant profiles, through cephalometric computerized analysis (Ricketts and McNamara)]. AB - This research aimed at establishing normative values, evaluating sexual dimorphism between cephalometric measurements, and correlating averages for the evaluation of biological compensations that act in the morphology of the facial soft tissue profile; for these purposes, the computerized cephalometric analyses of Ricketts and McNamara were employed. The sample comprised 40 lateral cephalometric radiographs of individuals with aesthetically pleasant profiles. The radiographs were analyzed through the Radiocef program 2.0, following the indicated cephalometric methods, and, later, Student's t-test and the coefficient of correlation of Pearson were used. For men, the cranial anterior length, upper lip length, effective midfacial length, effective mandibular length, lower anterior facial height and lower pharynx diameter presented significant higher dimensions. For women, the higher averages found were for the lower incisor extrusion and inferior lip protrusion. The masculine sample exhibited a smaller lower anterior facial height than the standardized averages by linear gauging because of the smallest effective mandibular length in this group; the same correlation was not significant for angular values. In both samples the inferior lip protrusion was directly related to the different spatial position of the upper and lower incisors, in their relation with the soft tissues and respective bony bases. PMID- 12908057 TI - Superinfecting microorganisms in patients under treatment with cyclosporin-A and its correlation to gingival overgrowth. AB - The aim of this study was to identify the presence of superinfecting microorganisms (Gram-negative enteric rods and Candida sp.) in heart transplant patients and correlate this with gingival overgrowth. Thirty patients (10 females, 20 males--mean age 45 years) were examined. All were under cyclosporin-A (CsA) therapy. Patients who had taken any antibiotics 3 months prior the study or had been submitted to periodontal therapy were not enrolled. Patients were required to have at least 6 teeth. The plaque index (PI), gingival index (GI), pocket depth (PD) and clinical attachment level (CAL) were recorded. Microbiological samples were taken from sulcus/pocket (s/p) and from stimulated saliva (ss) and submitted to analysis. Patients were divided into two groups: the ones with gingival overgrowth (GO) and those without gingival overgrowth (WGO). After statistical analysis (chi-square test, Student's t-test, Fisher test, p < or = 0.05), we concluded that there was no statistical difference between groups in the parameters of gender, CsA dosage, time since transplantation, PI, GI, PD and CAL. Gram-negative rods from either the sulcus/pocket or saliva samples were not found. Candida sp. was detected (s/p-30% and ss-30%). Stimulated saliva samples analysis determined that the presence of Candida sp. was associated with patients without gingival overgrowth. PMID- 12908058 TI - Smoking influences on the thickness of marginal gingival epithelium. AB - Smoking patients show reduction of inflammatory clinical signs that might be associated with local vasoconstriction and an increased gingival epithelial thickness. The purpose of this work was to evaluate the thickness of the marginal gingival oral epithelium in smokers and non-smokers, with clinically healthy gingivae or with gingivitis. Twenty biopsies were obtained from four different groups. Group I: non-smokers with clinically healthy gingivae (n = 5). Group II: non-smokers with gingivitis (n = 5). Group III: smokers with clinically healthy gingivae (n = 5). Group IV: smokers with gingivitis (n = 5). These biopsies were histologically processed, serially sectioned at 5 microns, stained with H. E., and examined by image analysis software (KS400), which was used to perform the morphometric evaluation and the quantification of the major epithelial thickness, the epithelial base thickness and the external and internal epithelial perimeters. Differences between the four groups were analyzed using ANOVA test and Tukey's test. The criteria for statistical significance were accepted at the probability level p < 0.05. A greater epithelial thickness was observed in smokers independent of the gingival health situation. PMID- 12908060 TI - Effects of ligature-induced periodontitis in pregnant Wistar rats. AB - The aim of this study was to evaluate the influence of ligature-induced periodontal disease in pregnant rats on their newborn's health parameters. Twenty four female adult Wistar rats were divided into two groups: the control group (G1) and the group that was submitted to dental ligatures around second upper molars (G2). After the four week period of development of periodontitis, the female animals were mated with male adult Wistar rats. There were no differences in the body weight of females between the two groups during mating and pregnancy. No differences were observed among the groups in relation to the viable newborn index. However, there were differences in newborn birth weight, explained by the diverse size of the litters. In this study, ligature-induced periodontal disease did not promote changes during pregnancy that resulted in low birth weight in newborn Wistar rats. PMID- 12908059 TI - Immunosuppressant therapy and bone loss in ligature-induced periodontitis--a study in rats. AB - Immunosuppressive agents have been recognized as a factor affecting the soft tissues of the periodontium. However, little is known about their effect on periodontitis progression. The aim of the present study was to investigate the influence of cyclosporin A (CsA) administration, associated or not with nifedipine, on the bone loss resulting from a ligature-induced periodontitis in rats. Twenty-four adult male Wistar rats were used. After anesthesia, the mandibular first molar was randomly assigned to receive the cotton ligature in the sulcular area while the contralateral tooth was left unligated. The animals were randomly assigned to one of the following treatments: Group A--saline solution; Group B--CsA (10 mg/kg); Group C--nifedipine (50 mg/kg); Group D--CsA (10 mg/kg) plus nifedipine (50 mg/kg). Forty-five days later, the animals were sacrificed and the specimens routinely processed for serial decalcified sections. Intergroup analysis did not reveal significant differences regarding the bone loss volume in the ligated teeth between the experimental treatments (0.46 +/- 0.11, 0.63 +/- 0.32, 0.53 +/- 0.14, 0.50 +/- 0.18, for groups A, B, C and D, respectively--p > 0.05). However, intragroup analysis showed a greater bone loss volume in the ligated teeth than in the unligated ones (p < 0.05). Within the limits of the present study, the conclusion was that the administration of CsA, associated or not with nifedipine, may not influence bone loss in ligature induced periodontitis in rats. PMID- 12908061 TI - Methodological considerations on descriptive studies of induced periodontal diseases in rats. AB - The aim of this study was to show the technique and the methodological approach used in describing histological characteristics of induced periodontal disease in rats. To reach that inflammatory process, periodontal disease was induced by ligature, with or without sucrose-rich diet. Twenty-four female adult (60 days old) Wistar rats were divided in four groups: Group 1, or control (which received standard diet), Group 2 (which received ligature around the upper second molars and a standard diet), Group 3 (which received a sucrose-rich diet), and Group 4 (which received ligature around the upper second molars and a sucrose-rich diet). The animals were followed for a period of 30 days, after which they were sacrificed. The upper second molars were removed, processed, and the histological characteristics were analyzed by a descriptive dichotomous method. The results were analyzed by the Fisher's exact test (significance level of 95%) and by a residual test, which showed the relation between groups and histological characteristics. The animals which received ligature (Groups 2 and 4) showed histological characteristics related with periodontitis, whilst the animals without ligatures showed no periodontal destruction. This was shown by a distribution of these groups in extremes of a graphic representation. The use of a ligature, as done in this study, was able to promote a chronic inflammatory process in the periodontium of rats, regardless of the adopted diet. The correspondence factorial analysis was capable of showing these characteristics, being one more tool to be used in histological research. PMID- 12908063 TI - Apical and periapical repair of dogs' teeth with periapical lesions after endodontic treatment with different root canal sealers. AB - The aim of this study was to evaluate the apical and periapical repair after root canal treatment of dogs' teeth with pulp necrosis and chronic periapical lesion using different root canal sealers. After periapical lesion induction, forty-four root canals of 3 dogs were submitted to biomechanical preparation using 5.25% sodium hypochlorite as an irrigating solution. A calcium hydroxide dressing (Calen PMCC) was applied for 15 days and the root canals were filled using the lateral condensation technique with gutta-percha points and Sealapex, AH Plus or Sealer Plus for sealing. After 180 days, the animals were sacrificed by anesthetic overdose and the obtained histological sections were stained with hematoxylin-eosin for optical microscopic analysis of the apical and periapical repair. The groups filled with Sealapex and AH Plus had better histological repair (p < 0.05) than the group filled with Sealer Plus, that had unsatisfactory results. PMID- 12908064 TI - In vitro evaluation of different chemical agents for the decontamination of gutta percha cones. AB - This study evaluated the effectiveness of three disinfectants used in Dentistry for decontamination of gutta-percha cones. Sixty gutta-percha cones were contaminated with standardized pure cultures of five species of microorganisms (Enterococcus faecalis ATCC 29212, Staphylococcus aureus ATCC 25923, Candida albicans ATCC CBS-ICB/USP 562, Bacillus subtilis spores ATCC 6633 and Streptococcus mutans ATCC 25175). The cones were treated with 10% polyvinylpyrrolidone-iodine aqueous solution (PVP-I; Groups 1 and 2), 5.25% aqueous sodium hypochlorite (Groups 3 and 4) and paraformaldehyde tablets (Group 5). All chemical agents were efficient for the cold sterilization of gutta-percha cones in short time periods. PMID- 12908065 TI - Evaluation of the substantivity of chlorhexidine in association with sodium fluoride in vitro. AB - The efficacy of the fluoride-chlorhexidine association in the prevention of gingivitis and caries has been advocated for a number of years. The objective of the association of these therapeutic agents is a synergistic action. The aim of the present study was to determine the substantivity of chlorhexidine associated or not to sodium fluoride at different intervals of time, in vitro. Bovine enamel surfaces were treated with 0.12% chlorhexidine gluconate (Periogard--Colgate) or 0.05% sodium fluoride with 0.12% chlorhexidine (Duplak--Dentsply) solutions for one minute. Fragments were placed in distilled water and samples were collected at intervals of 5, 30 and 360 minutes and analyzed by spectrophotometry in the visible ultraviolet region. Substantivity was evaluated by the measurement of chlorhexidine desorption from the treated slabs. The mean values obtained were statistically analyzed by Student's t-test. The results showed that the concentration of chlorhexidine decreased when it was used in association with sodium fluoride. The difference between the groups was statistically significant (p < 0.05). These in vitro results suggest that the association between sodium fluoride and chlorhexidine in the studied concentrations appears to have no beneficial effect because of the decrease in the substantivity of chlorhexidine. PMID- 12908062 TI - Effect of prophylactic treatments on the superficial roughness of dental tissues and of two esthetic restorative materials. AB - Dental prophylaxis is a common way to remove dental plaque and stain, both undesirable factors in most dentistry procedures. However, besides cleaning the tooth surface, prophylactic techniques may increase the surface roughness of restorations and dental tissues, which, in turn, may result in plaque accumulation, superficial staining and superficial degradation. This study evaluated the effect of three prophylactic techniques--sodium bicarbonate jet, pumice paste and whiting paste--on the superficial roughness of two restorative materials--a composite resin and a compomer--and on the superficial roughness of two dental surfaces--enamel and cementum/dentin--through rugosimetric and scanning electron microscopy (SEM) analysis. Statistical analysis of the rugosimetric data showed that the use of pumice paste on enamel produced a significantly smoother surface than the natural surface. However, comparing the effect of the three techniques, prophylaxis with the pumice paste produced a rougher surface than did the other techniques as regards enamel and cementum/dentin probably due to its abrasiveness. On composite resin, the pumice paste only produced a rougher surface than did the whiting paste. On compomer, all of the applied treatments produced similar results. Based on rugosimetric and SEM analysis, we could conclude that the prophylactic treatments employed did not improve roughness of the studied surfaces. As to the effects of the techniques, they were different depending on the surfaces on which the prophylactic treatments were applied. PMID- 12908066 TI - [Collagen type I expression in squamous cell carcinoma of the oral cavity]. AB - An analysis of the extracellular matrix at the invasive front of squamous cell carcinoma of the oral cavity may improve the understanding of tumour cell matrix interactions during malignancy growth. Alterations in collagen I expression may influence cellular invasion and metastasis. In this work, 23 cases of squamous cell carcinoma were submitted to the Anneroth's malignancy grading system. H. E. and sirius red staining were used. Immuno-histochemical expression of collagen type I protein was observed in different malignancy scores. As a result, it was observed that the extracellular matrix in squamous cell carcinoma of the oral cavity shows different patterns of collagen I expression in low and high scores of malignancy. PMID- 12908067 TI - [The frequency of Candida sp. in biopsies of oral mucosal lesions]. AB - Candidosis is the most common fungal infection in the oral cavity, and is usually associated with local and systemic predisposing factors. The occurrence and relevance of Candidal infection in oral lesions such as liquen planus, leukoplakias and carcinomas are still to be understood. The aim of the present study was to define the frequency of infection by Candida sp. on biopsies of oral mucosal lesions and associate its presence with malignant and dysplastic lesions. Histopathology reports issued between 1990 and 2001 inclusive were reviewed. Three sections of each mucosal biopsy were stained using the periodic acid-Schiff (PAS) technique. From the 832 biopsies 27.2% were PAS positive, of which 83.25% were obtained from male patients. There was positive association between fungic infection and mild, moderate and severe epithelial dysplasia, squamous cell carcinoma and hiperqueratosis (p < 0.05). There was no association between fungic infection and inflammatory fibrous hyperplasia, hyperkeratosis, lichen planus and pyogenic granuloma (p < 0.05). The frequency of infection in the tongue was significantly higher (p < 0.05) than in the other sites. Our results do not show a causal relation between Candida sp. and dysplastic lesions and carcinomas, but do confirm the higher presence of that microrganism in those lesions. PMID- 12908068 TI - Homeobox genes: a molecular link between development and cancer. AB - Homeobox genes are regulatory genes encoding nuclear proteins that act as transcription factors, regulating aspects of morphogenesis and cell differentiation during normal embryonic development of several animals. Vertebrate homeobox genes can be divided in two subfamilies: clustered, or HOX genes, and nonclustered, or divergent, homeobox genes. During the last decades, several homeobox genes, clustered and nonclustered ones, were identified in normal tissue, in malignant cells, and in different diseases and metabolic alterations. Homeobox genes are involved in the normal teeth development and in familial teeth agenesis. Normal development and cancer have a great deal in common, as both processes involve shifts between cell proliferation and differentiation. The literature is accumulating evidences that homeobox genes play an important role in oncogenesis. Many cancers exhibit expression of or alteration in homeobox genes. Those include leukemias, colon, skin, prostate, breast and ovarian cancers, among others. This review is aimed at introducing readers to some of the homeobox family functions in normal tissues and especially in cancer. PMID- 12908069 TI - Coinfection with Mycoplasma pneumoniae and Chlamydia pneumoniae in ruptured plaques associated with acute myocardial infarction. AB - OBJECTIVE: To study atheromas, Mycoplasma pneumoniae (M. pneumoniae), and Chlamydia pneumoniae (C. pneumoniae). METHODS: C. pneumoniae was studied with immunohistochemistry and M. pneumoniae with in situ hybridization (ISH), in segments of coronary arteries (SCA) as follows: group A - thrombosed ruptured plaques (TRP) of 23 patients who died due to acute myocardial infarction (AMI); group B - 23 nonruptured plaques (NRP) of group A patients; group C - NRP of 11 coronary patients who did not die due to AMI; and group D - 11 SCA from patients with dilated cardiomyopathy or Chagas' disease without atherosclerosis. RESULTS: The mean number of C. pneumoniae+ cells/400x in groups A, B, C, and D was, respectively, 3.3 +/- 3.6; 1.0 +/- 1.3; 1.2 +/- 2.4; and 0.4 +/- 0.3; and the percentage of M. pneumoniae area was, respectively, 3.9 +/- 3.5; 1.5 +/- 1.6; 0.9 +/- 0.9; and 0.4 +/- 0.2. More M. pneumoniae and C. pneumoniae were found in of group A than in group B (P<0.01). Good correlation was seen between the area of the vessel and the M. pneumoniae area in the plaque (r = 0.46; P=0.001) and between C. pneumoniae+ cells and CD4+ T lymphocytes (r = 0.42; P<0.01). The number of C. pneumoniae+ cells correlated with CD20+ B cells (r=0.48; P<0.01). CONCLUSION: M. pneumoniae and C. pneumoniae are more frequently found in TRP correlate with the intensity of the inflammation and diameter of the vessel (positive remodeling). PMID- 12908071 TI - Cellular cholesterol efflux mediated by HDL isolated from subjects with low HDL levels and coronary artery disease. AB - OBJECTIVE: The aim of this study was to verify whether HDL particles isolated from patients with coronary artery disease (CAD) and low HDL-C had diminished ability to promote cholesterol efflux from cultured cells compared with HDL isolated from subjects without CAD and with normal HDL-C. METHODS: Smooth muscle cells isolated from human aortas cultured and radiolabeled with H-cholesterol were loaded with cholesterol and incubated with increasing concentrations of HDL isolated from 13 CAD patients with low HDL-C (CAD group) or from 5 controls without CAD (C group). Efflux of cellular cholesterol was measured by cellular depletion of radiolabeled cholesterol and by the appearance of H-cholesterol into experimental medium expressed as a percentage of total labeled cholesterol. RESULTS: Cholesterol efflux increased with the amount of HDL present in the medium, and no difference was found between groups at various HDL protein concentrations: efflux was 28 +/- 6.3% (C) and 25.5 +/- 8.9% (CAD) with 25 microg/mL; 34 +/- 4.3% (C) and 31.9 +/- 6.6% (CD) with 50 micro g/mL and 39.5 +/- 3.5% (C) and 37.1 +/- 4.4% (CAD) with 100 micro g/mL, HDL. CONCLUSION: Because the HDL fraction of CAD patients with low HDL-C have normal ability to extract cholesterol from cells of the vessel wall, it is suggested that low HDL-C atherogenicity should be ascribed to diminished concentrations of HDL particles rather than to the qualitative properties of the HDL fraction. PMID- 12908070 TI - Characterization of the in vivo cardiac electrophysiologic effects of high-dose cocaine in closed-chest, anesthetized dogs with normal hearts. AB - OBJECTIVE: To characterize the cardiac electrophysiologic effects of cocaine. METHODS: In 8 dogs (9-13 kg), electrophysiologic parameters and programmed stimulation were undertaken using transvenous catheters at baseline, and after cocaine intravenous infusion (12 mg/kg bolus followed by 0.22 mg/kg/min for 25 minutes). RESULTS: Cocaine plasma levels (n=5) rose to 6.73 +/- 0.56 mg/mL. Cocaine did not affect sinus cycle length and arterial pressure. Cocaine prolonged P wave duration (54 +/- 6 vs 73 +/- 4 ms, P<0.001), PR interval (115 +/ 17 vs 164 +/- 15 ms, P<0.001), QRS duration (62 +/- 10 vs 88 +/- 14 ms, P<0.001), and QTc interval (344 +/- 28 vs 403 +/- 62 ms, P=0.03) but not JT interval (193 +/- 35 vs 226 +/- 53 ms, NS). Cocaine prolonged PA (9 +/- 6 vs 23 +/- 8 ms, P<0.001), AH (73 +/- 16 vs 92 +/- 15 ms; P=0.03), and HV (35 +/- 5 vs 45 +/- 3 ms; P<0.001) intervals and Wenckebach point (247 +/- 26 vs 280 +/- 28 ms, P=0.04). An increase occurred in atrial (138 +/- 8 vs 184 +/- 20 ms; P<0.001) and ventricular (160 +/- 15 vs 187 +/- 25 ms; P=0.03) refractoriness at a cycle length of 300 ms. Atrial arrhythmias were not induced in any dog. Ventricular fibrillation (VF) was induced in 2/8 dogs at baseline and 4/8 dogs after cocaine. CONCLUSION: High doses of cocaine exert significant class I effects and seem to enhance inducibility of VF but not of atrial arrhythmias. PMID- 12908072 TI - A new proposal to guide velocity and inclination in the ramp protocol for the treadmill ergometer. AB - OBJECTIVE: To suggest criteria to guide protocol prescription in ramp treadmill testing, according to sex and age, based on velocity, inclination, and max VO2 reached by the population studied. METHODS: Prospective study describing heart rate (HR), time, velocity, inclination, and VO2 estimated at maximum effort of 1840 individuals from 4 to 79 years old, who performed a treadmill test (TT) according to the ramp protocol. A paired Student t test was used to assess the difference between predicted and reached max VO2, calculated according to the formulas of the "American College of Sports Medicine". RESULTS: Submaximal HR was surpassed in 90.1% of the examinations, with a mean time of 10.0 2.0 minute. Initial and peak inclination velocity of the exercise and max VO2 were inversely proportional to age and were greater in male patients. Predicted Max VO2 was significantly lower than that reached in all patients, except for female children and adolescents (age < 20 years old). CONCLUSION: Use of velocity, inclination, and maximum VO2 actually reached, as a criterion in prescribing the ramp protocol may help in the performance of exercise in treadmill testing. The ramp protocol was well accepted in all age groups and sexes with exercise time within the programmed 8 to 12 minutes. PMID- 12908073 TI - Comparative study of the use of diltiazem as an antispasmodic drug in coronary angiography via the transradial approach. AB - OBJECTIVE: To evaluate the impact of the use, prior to the procedure, of injectable diltiazem to prevent complications. METHODS: Between September 2000 and July 2001, 50 patients underwent transradial coronary angiography and were randomized to receive placebo (GI) or diltiazem (GII) through a catheter inserted into the radial artery. All patients received isosorbide mononitrate. Ultrasound analyses of the radial artery were performed before examination, 30 minutes afterwards, and 7 days afterwards to evaluate the flow, the diameter, and the artery output. RESULTS: The radial artery diameter of GI was 2.4d +/- 0.5 mm before the procedure and 2.3 +/- 0.5 mm after 30 minutes (NS), whereas in GII the diameter was 2.2 +/- 0.3 mm before the examination and +/- 2.5 0.4 mm 30 minutes after it (P<0.001). Radial artery output in group 1 was 7.3 +/- 5.l2 mL/min before the examination and 6.1 +/- 3.5 mL/min 30 minutes after the examination (NS), and GII had an increase of 5.9 +/- 2.5 mL/min before examination to 9.05 +/ 7.78 mL/min after the examination (P=0.04). Complications (spasm, occlusion, and partial obstruction) occurred in 4 patients (17.4%) in GI and did not occur in GII (P=0.04). CONCLUSION: The study suggests a decrease in vascular complications through the transradial access for coronary angiography with the use of diltiazem as an antispasmodic drug, resulting in the significant increase in the diameter of the radial artery and radial artery output. PMID- 12908074 TI - Enalapril alters the formation of the collagen matrix in spontaneously hypertensive rats. AB - OBJECTIVE: To assess the effect of the inhibition of the angiotensin-converting enzyme on the collagen matrix (CM) of the heart of newborn spontaneously hypertensive rats (SHR) during embryonic development. METHODS: The study comprised the 2 following groups of SHR (n=5 each): treated group - rats conceived from SHR females treated with enalapril maleate (15 mg. kg-1.day-1) during gestation; and nontreated group - offspring of nontreated females. The newborns were euthanized within the first 24 hours after birth and their hearts were removed and processed for histological study. Three fields per animal were considered for computer-assisted digital analysis and determination of the volume densities (Vv) of the nuclei and CM. The images were segmented with the aid of Image Pro Plus 4.5.029 software (Media Cybernetics). RESULTS: No difference was observed between the treated and nontreated groups in regard to body mass, cardiac mass, and the relation between cardiac and body mass. A significant reduction in the Vv[matrix] and a concomitant increase in the Vv[nuclei] were observed in the treated group as compared with those in the nontreated group. CONCLUSION: The treatment with enalapril of hypertensive rats during pregnancy alters the collagen content and structure of the myocardium of newborns. PMID- 12908076 TI - Marfan's syndrome: early and severe form in siblings. AB - Marfan's syndrome is an inherited disorder of the connective tissue. Cardiologic manifestations, especially aortic dilation, are important causes of morbidity and mortality in the clinical course of the disease in adults and teenagers. In children, the presence of aortic aneurysm and its dissection or rupture is rare, occurring in patients with genetic mutation of the fibrillin gene but not in those who have the familial form of the disease. We describe here 2 patients, from the same family (siblings), diagnosed with gigantic aortic aneurysm early in infancy, one of them successfully undergoing surgery. PMID- 12908075 TI - Detection of incipient left ventricular hypertrophy in mild to moderate arterial hypertension with normal electrocardiogram and echocardiogram: a new use for signal-averaged electrocardiography. AB - OBJECTIVE: To assess signal-averaged electrocardiogram (SAECG) for diagnosing incipient left ventricular hypertrophy (LVH). METHODS: A study with 115 individuals was carried out. The individuals were divided as follows: GI - 38 healthy individuals; GII - 47 individuals with mild to moderate hypertension and normal findings on echocardiogram and ECG; and GIII - 30 individuals with hypertension and documented LVH. The magnitude vector of the SAECG was analyzed with the high-pass cutoff frequency of 40 Hz through the bidirectional four-pole Butterworth high-pass digital filter. The mean quadratic root of the total QRS voltage (RMST) and the two-dimensional integral of the QRS area of the spectro temporal map were analyzed between 0 and 30 Hz for the frequency domain (Int FD), and between 40 and 250 Hz for the time domain (Int TD). The electrocardiographic criterion for LVH was based on the Cornell Product. Left ventricular mass was calculated with the Devereux formula. RESULTS: All parameters analyzed increased from GI to GIII, except for Int FD (GII vs GIII) and RMST log (GII vs GIII). Int TD showed greater accuracy for detecting LVH with an appropriate cutoff > 8 (sensitivity of 55%, specificity of 81%). Positive values (> 8) were found in 56.5% of the G II patients and in 18.4% of the GI patients (p< 0.0005). CONCLUSION: SAECG can be used in the early diagnosis of LVH in hypertensive patients with normal ECG and echocardiogram. PMID- 12908077 TI - Malignant hypertension and hypertensive encephalopathy in primary aldosteronism caused by adrenal adenoma. AB - Two cases are reported as follows: 1) 1 female patient with accelerated-malignant hypertension secondary to an aldosterone-producing adrenal adenoma; and 2) 1 female patient with adrenal adenoma, severe hypertension, and hypertensive encephalopathy. This association is a rare clinical finding, and malignant hypertension may modify the hormonal characteristic of primary aldosteronism, making its diagnosis more difficult. The diagnosis of primary aldosteronism should be considered in patients with malignant hypertension or hypertensive encephalopathy if persistent hypokalemia occurs. Identification of primary aldosteronism is of paramount importance for the patient's evolution, because the surgical treatment makes the prognosis more favorable. PMID- 12908078 TI - [Clinico-radiographic correlation. Case 7/2003 - Instituto do Coracao do Hospital das Clinicas da FMUSP]. PMID- 12908079 TI - Evidence-based medicine. A lesson for electrocardiography? PMID- 12908080 TI - High-efficiency biolistic co-transformation and regeneration of 'Chardonnay' (Vitis vinifera L.) containing npt-II and antimicrobial peptide genes. AB - A reliable and efficient system for transformation and regeneration of 'Chardonnay' (Vitis vinifera L.) plants via microprojectile bombardment was developed. Improvements over the previous biolistic transformation system included: (1) the use of gold particles for bombardment; (2) step-wise selection at 10 then 15 mg/l kanamycin; and (3) embryo induction at 27 degrees C. Embryogenic cell cultures were either bombarded with pBI426, which contains the reporter gene gus (uidA) coding for beta-glucuronidase (GUS), or were co bombarded with pSAN237 carrying the npt-II (neomycin phosphotransferase II) selectable marker gene, and a second plasmid with an antimicrobial peptide gene. A large number of transient (7,883 +/- 1,928) and stable (46 +/- 32) blue spots per plate at 2 and 95 days after bombardment, respectively, were obtained according to GUS expression analyses. A total of 447 putative transgenic embryos was harvested from 84 bombarded plates. From these embryos, 242 (54%) were regenerated into plants within the first year of the experiment. Southern blot analyses confirmed integration of the transgenes into the grape genome. Co transformation was tested with four separate antimicrobial constructs. The co transformation frequency of unlinked genes was 48% as measured by polymerase chain reaction (PCR), and 56% as estimated by dot blot hybridization. Expression of the gus gene, and PCR and Southern blot analyses of npt-II and antimicrobial genes from regenerated plants document stable transformation of 'Chardonnay' and establish the parameters for highly-efficient biolistic transformation in V. vinifera. PMID- 12908081 TI - Sugar acts as a regulatory signal on the wound-inducible expression of SbHRGP3::GUS in transgenic plants. AB - SbHRGP3 encodes an HRGP whose expression is correlated with the cessation of root elongation in soybean. The wound-inducible expression of SbHRGP3 interestingly requires sucrose although wounding alone induces the expression of many HRGP genes. To examine whether sugar serves as a specific signal on the wound inducible expression or whether sugar is required to provide ATP, we examined SbHRGP3:: GUS expression in transgenic tobacco plants. Various oligosaccharides including non-metabolizable sugar induced SbHRGP3:: GUS expression in transgenic plants. The inhibitors of photosynthesis and of cellular respiration did not affect the wound-inducible expression of SbHRGP3:: GUS. However, the induction was significantly affected by PCMBS, an inhibitor of active apoplastic phloem loading of sucrose, suggesting that SbHRGP3:: GUS expression in phloem tissues requires translocated sucrose. We therefore propose that sugar acts as a specific regulatory signal on the wound-inducible expression of SbHRGP3, rather than acting as a simple provider of ATP. PMID- 12908082 TI - Induction of TNF-alpha, uPA, IL-8 and MCP-1 by doxorubicin in human lung carcinoma cells. AB - PURPOSE: We have previously demonstrated doxorubicin-induced urokinase (uPA) and interleukin-8 (IL-8) expression in human H69 small-cell lung carcinoma (SCLC) cells by a microarray technique using Human Cancer Chip version 2, in which 425 human "cancer-related" genes are spotted on the plates. The microarray analysis also revealed a significant induction of tumor necrosis factor-alpha (TNF-alpha), and doxorubicin-induced macrophage chemoattractant protein-1 (MCP-1) expression was demonstrated by an RNase protection assay. We extended the study by testing the effects of doxorubicin on the induction of TNF-alpha, uPA, IL-8 and MCP-1 in other types of lung carcinoma cells. METHODS: We investigated the effects of doxorubicin on the expression of TNF-alpha, uPA, IL-8 and MCP-1 in 12 human lung carcinoma cell lines, including five SCLC, three adenocarcinoma and four squamous cell carcinoma cells. The surface expression of their receptors was also investigated. RESULTS: TNF-alpha was significantly induced in three cell lines, H69, SBC-7 (SCLC) and PC-9 (adenocarcinoma), uPA in five cell lines, H69, SBC-7, EBC-1 (squamous cell), EBC-2 (squamous cell), and Sq-1 (squamous cell), IL-8 in three cell lines, H69, PC-9 and EBC-1, and MCP-1 in five cell lines, H69, SBC-3 (SCLC), SBC-7, PC-9 and Sq-1. In H69 cells, TNF-alpha antigen levels were increased approximately fivefold in the conditioned medium of doxorubicin-treated cells, in parallel with an increase in mRNA levels. As with uPA and IL-8, the maximum induction was observed at the "sublethal" concentrations of 2 and 4 microM at which cell growth was slightly inhibited 24 h after treatment. Furthermore, the cells did not express receptors including types I and II TNF alpha receptors, uPA receptor (uPAR), C-x-C-chemokine receptor-1 (CXCR-1), or C-C chemokine receptor-2, corresponding to TNF-alpha, uPA, IL-8 and MCP-1, respectively, that were induced by doxorubicin in the cells, although SBC-7 cells expressed uPAR, and EBC-1 cells expressed CXCR-1. CONCLUSIONS: TNF-alpha, uPA, IL 8 and MCP-1 induced and secreted from tumor cells upon doxorubicin stimulation may activate surrounding cells expressing the receptors such as neutrophils and monocytes/macrophages in a paracrine fashion. TNF-alpha is a major proinflammatory cytokine, and IL-8 and MCP-1 are major chemoattractants for neutrophils and monocytes/macrophages, respectively. Furthermore, uPA activates matrix metalloproteinase 9 which can truncate and activate IL-8. Thus, the simultaneous induction of TNF-alpha, uPA, IL-8 and MCP-1 may enhance the interaction between tumor and inflammatory/immune cells, and augment cytotoxicity. PMID- 12908083 TI - Improved method for determination of ammonia and nitrite oxidation activities in mixed bacterial cultures. AB - A simple and reliable method to measure the activity of ammonia and nitrite oxidisers in mixed bacterial cultures was developed. The developed method differentiates between the ammonia and nitrite oxidisers by consecutive injection of NO2- and NH4+. The main advantage of this method is that it avoids the use of metabolic inhibitors for ammonia or nitrite oxidisers, as used by other methods. Moreover, it allows measuring of the short-term effect of an inhibitor on both the ammonia and nitrite oxidisers in one test under controlled environmental conditions (pH, temperature). The developed method was applied to determine the inhibitory effects of salt (NaCl up to 15 g Cl/l) on an enriched culture of nitrifying bacteria. The results of the method demonstrate its potential to accurately determine the individual activities of nitrite and ammonia oxidisers. PMID- 12908084 TI - Microbial fed-batch production of 1,3-propanediol by Klebsiella pneumoniae under micro-aerobic conditions. AB - The microbial production of 1,3-propanediol (1,3-PD) by Klebsiella pneumoniae under micro-aerobic conditions was investigated in this study. The experimental results of batch fermentation showed that the final concentration and yield of 1,3-PD on glycerol under micro-aerobic conditions approached values achieved under anaerobic conditions. However, less ethanol was produced under microaerobic than anaerobic conditions at the end of fermentation. The batch micro-aerobic fermentation time was markedly shorter than that of anaerobic fermentation. This led to an increment of productivity of 1,3-PD. For instance, the concentration, molar yield, and productivity of 1,3-PD of batch micro-aerobic fermentation by K. pneumoniae DSM 2026 were 17.65 g/l, 56.13%, and 2.94 g l(-1) h(-1), respectively, with a fermentation time of 6 h and an initial glycerol concentration of 40 g/l. Compared with DSM 2026, the microbial growth of K. pneumoniae AS 1.1736 was slow and the concentration of 1,3-PD was low under the same conditions. Furthermore, the microbial growth in fed-batch fermentation by K. pneumoniae DSM 2026 was faster under micro-aerobic than anaerobic conditions. The concentration, molar yield, and productivity of 1,3-PD in fed-batch fermentation under micro-aerobic conditions were 59.50 g/l, 51.75%, and 1.57 g l(-1) h(-1), respectively. The volumetric productivity of 1,3-PD under microaerobic conditions was almost twice that of anaerobic fed-batch fermentation, at 1.57 and 0.80 g l(-1) h(-1), respectively. PMID- 12908085 TI - Isolation of microorganisms for biological detoxification of lignocellulosic hydrolysates. AB - Acid pretreatment of lignocellulosic biomass releases furan and phenolic compounds, which are toxic to microorganisms used for subsequent fermentation. In this study, we isolated new microorganisms for depletion of inhibitors in lignocellulosic acid hydrolysates. A sequential enrichment strategy was used to isolate microorganisms from soil. Selection was carried out in a defined mineral medium containing a mixture of ferulic acid (5 mM), 5-hydroxymethylfurfural (5 HMF, 15 mM), and furfural (20 mM) as the carbon and energy sources, followed by an additional transfer into a corn stover hydrolysate (CSH) prepared using dilute acid. Subsequently, based on stable growth on these substrates, six isolates- including five bacteria related to Methylobacterium extorquens, Pseudomonas sp, Flavobacterium indologenes, Acinetobacter sp., Arthrobacter aurescens, and one fungus, Coniochaeta ligniaria--were chosen. All six isolates depleted toxic compounds from defined medium, but only C. ligniaria C8 (NRRL 30616) was effective at eliminating furfural and 5-HMF from CSH. C. ligniaria NRRL 30616 may be useful in developing a bioprocess for inhibitor abatement in the conversion of lignocellulosic biomass to fuels and chemicals. PMID- 12908086 TI - A derivative of the menaquinone precursor 1,4-dihydroxy-2-naphthoate is involved in the reductive transformation of carbon tetrachloride by aerobically grown Shewanella oneidensis MR-1. AB - Transformation of carbon tetrachloride (CT) by Shewanella oneidensis MR-1 has been proposed to involve the anaerobic respiratory-chain component menaquinone. To investigate this hypothesis a series of menaquinone mutants were constructed. The menF mutant is blocked at the start of the menaquinone biosynthetic pathway. The menB, menA and menG mutants are all blocked towards the end of the pathway, being unable to produce 1,4-dihydroxy-2-naphthoic acid (DHNA), demethyl menaquinone and menaquinone, respectively. Aerobically grown mutants unable to produce the menaquinone precursor DHNA (menF and menB mutants) showed a distinctly different CT transformation profile than mutants able to produce DHNA but unable to produce menaquinone (menA and menG mutants). While DHNA did not reduce CT in an abiotic assay, the addition of DHNA to the menF and menB mutants restored normal CT transformation activity. We conclude that a derivative of DHNA, that is distinct from menaquinone, is involved in the reduction of CT by aerobically grown S. oneidensis MR-1. When cells were grown anaerobically with trimethylamine-N-oxide as the terminal electron acceptor, all the menaquinone mutants showed wild-type levels of CT reduction. We conclude that S. oneidensis MR-1 produces two different factors capable of dehalogenating CT. The factor produced under anaerobic growth conditions is not a product of the menaquinone biosynthetic pathway. PMID- 12908087 TI - Sensitivity of chronically HIV-1 infected HeLa cells to electrical stimulation. AB - Use of combination anti-retroviral drug regimens including protease inhibitors dramatically decreased morbidity and mortality rates in HIV-1 infected individuals. However, such combination therapies appear to have many side effects, in addition to the emergence of resistant HIV-1 strains. Therefore, in this study we sought to elucidate novel therapeutic principles against HIV-1 infection. We examined the effects of electrical stimulation on both chronically HIV-1(LAI) infected HeLa cells (P6 HeLa/HIV-1(LAI)) and uninfected cells (P6 HeLa). Cells were cultured on an optically transparent electrode and application of potential at 1.0 V vs Ag/AgCl was performed over time periods ranging from 10 min to 60 min. Both P6 HeLa/HIV-1(LAI) and P6 HeLa cells were progressively damaged as the duration of electrical stimulation increased. However, P6 HeLa/HIV 1(LAl) cells were much more influenced by electrical stimulation than P6 HeLa cells. The difference in damage rate was most obvious at 30 min of electrical stimulation, with damaged cells accounting for about 87% and 4% of P6 HeLa/HIV 1(LAI) and P6 HeLa cells, respectively. After the application of potential for 20 min, the proliferation of P6 HeLa/HIV-1(LAI) cells was markedly inhibited, while the P6 HeLa cells proliferated to an extent similar to that of uninfected cells without application of potential. These results indicate that sensitivity to electrical stimulation is much higher in chronically HIV-1 infected cells than in uninfected cells. This could be considered as a useful new approach against HIV-1 infection. PMID- 12908088 TI - Enrichment of microbial community generating electricity using a fuel-cell-type electrochemical cell. AB - A fuel cell was used to enrich a microbial consortium generating electricity, using organic wastewater as the fuel. Within 30 days of enrichment the maximum current of 0.2 mA was generated with a resistance of 1 kOhms. Current generation was coupled to a fall in chemical oxygen demand from over 1,700 mg l(-1) down to 50 mg l(-1). Denaturing gradient gel electrophoresis showed a different microbial population in the enriched electrode from that in the sludge used as the inoculum. Electron microscopic observation showed a biofilm on the electrode surface and microbial clumps. Nanobacteria-like particles were present on the biofilm surface. Metabolic inhibitors and electron acceptors inhibited the current generation. 16S ribosomal RNA gene analysis showed a diverse bacterial population in the enrichment culture. These findings demonstrate that an electricity-generating microbial consortium can be enriched using a fuel cell and that the electrochemical activity is a form of anaerobic electron transfer. PMID- 12908089 TI - A close encounter with DNA. AB - Atomic force microscopy, associated with surface science, has the potential to resolve the secondary structure of DNA in liquid form with unusually high resolution and with unprecedented accuracy. PMID- 12908090 TI - Pinhole imaging of 131I-metaiodobenzylguanidine (131I-MIBG) in an animal model of neuroblastoma. AB - PURPOSE: To evaluate (131)I-MIBG scintigraphic localization of xenotransplanted and spontaneously arising neuroblastomas in murine models of high-risk neuroblastoma. METHODS: Neuroblastoma xenografts were created by inoculation of human neuroblastoma cell suspensions into the subcutaneous flanks of athymic nude mice. In addition, spontaneous paraspinal neuroblastomas were detected by direct palpation in MYCN transgenic mice. After measured tumor volumes exceeded 200 mm(3), each mouse received an intraperitoneal injection of 18 muCi/g (131)I metaiodobenzylguanidine ((131)I-MIBG). Pinhole scintigraphy was performed to evaluate the MIBG biodistribution and to attempt to visualize the tumors. Each mouse was imaged on a gamma camera equipped with a 3-mm pinhole on one head and an HEGP collimator on the other. RESULTS: Images demonstrated absorption of radiolabeled MIBG and visualization of tumors. Analysis of the images allowed for quantification of relative MIBG uptake and for determination of linear and area measurements of the tumors. CONCLUSION: High-energy pinhole imaging effectively demonstrates uptake of radiolabeled MIBG by human neuroblastoma tumors in murine laboratory models. This technique allows for in vivo assessment of tumor burden. In the future, we plan to use this method to evaluate sensitivity for detecting metastatic spread as well as investigating the therapeutic efficacy of high-dose (131)I-MIBG in combination with radiosensitizing agents. PMID- 12908092 TI - Diffusion-weighted imaging in acute bacterial meningitis in infancy. AB - Bacterial meningitis is frequently fatal or leads to severe neurological impairment. Complications such as vasculitis, resulting in infarcts, should be anticipated and dealt with promptly. Our aim was to demonstrate the complications of meningitis by diffusion weighted imaging (DWI) in patients who deteriorated despite therapy. We studied 13 infants between the ages of 1 day and 32 months who presented with symptoms ranging from fever and vomiting to seizures, encephalopathy and coma due to bacterial meningitis, performing MRI, including DWI, 2-5 days after presentation. Multiple infarcts were found on DWI in 12 of the 13, most commonly in the frontal lobes (in 10). Global involvement was seen in four children, three of whom died; the fourth had a very poor outcome. In one case abnormalities on DWI were due to subdural empyemas. We diagnosed vasculitis in three of five patients studied with MRA. We think DWI an important part of an MRI study in infants with meningitis. Small cortical or deep white-matter infarcts due to septic vasculitis can lead to tissue damage not easily recognized on routine imaging and DWI can be used to confirm that extra-axial collections represent empyemas. PMID- 12908094 TI - Evaluation of the combined measurement uncertainty in isotope dilution by MC-ICP MS. AB - The combination of metrological weighing, the measurement of isotope amount ratios by a multicollector inductively coupled plasma mass spectrometer (MC-ICP MS) and the use of high-purity reference materials are the cornerstones to achieve improved results for the amount content of lead in wine by the reversed isotope dilution technique. Isotope dilution mass spectrometry (IDMS) and reversed IDMS have the potential to be a so-called primary method, with which close comparability and well-stated combined measurement uncertainties can be obtained. This work describes the detailed uncertainty budget determination using the ISO-GUM approach. The traces of lead in wine were separated from the matrix by ion exchange chromatography after HNO(3)/H(2)O(2) microwave digestion. The thallium isotope amount ratio ( n((205)Tl)/ n((203)Tl)) was used to correct for mass discrimination using an exponential model approach. The corrected lead isotope amount ratio n((206)Pb)/ n((208)Pb) for the isotopic standard SRM 981 measured in our laboratory was compared with ratio values considered to be the least uncertain. The result has been compared in a so-called pilot study "lead in wine" organised by the CCQM (Comite Consultatif pour la Quantite de Matiere, BIPM, Paris; the highest measurement authority for analytical chemical measurements). The result for the lead amount content k(Pb) and the corresponding expanded uncertainty U given by our laboratory was:k(Pb)=1.329 x 10-10mol g-1 (amount content of lead in wine)U[k(Pb)]=1.0 x 10-12mol g-1 (expanded uncertainty U=kxuc, k=2)The uncertainty of the main influence parameter of the combined measurement uncertainty was determined to be the isotope amount ratio R(206,B) of the blend between the enriched spike and the sample. PMID- 12908093 TI - Endovascular treatment of brain-stem arteriovenous malformations: safety and efficacy. AB - Our purpose was to evaluate the safety and efficacy of endovascular treatment of brain-stem arteriovenous malformations (AVMs), reviewing six cases managed in the last 5 years. There were four patients who presented with bleeding, one with a progressive neurological deficit and one with obstructive hydrocephalus. Of the six patients, one showed 100%, one 90%, two 75% and two about 50% angiographic obliteration of the AVM after embolisation; the volume decreased about 75% on average. Five patients had a good outcome and one an acceptable outcome, with a mild postprocedure neurological deficit; none had further bleeding during midterm follow-up. Endovascular management of a brain-stem AVM may be an alternative to treatment such as radiosurgery and microsurgery in selected cases. It may be not as risky as previously thought. Embolisation can reduce the size of the AVM and possibly make it more treatable by radiosurgery and decrease the possibility of radiation injury. PMID- 12908096 TI - The prostaglandins epoprostenol and iloprost increase left ventricular contractility in vivo. AB - OBJECTIVE: The principal effects of prostaglandin I(2) are vasodilation and inhibition of platelet aggregation induced by a rise in the intracellular second messenger cAMP. In the heart a rise in intracellular myocardial cAMP increases contractility. We examined whether prostaglandin I(2) increases left ventricular contractility in vivo. The effects of epoprostenol and iloprost on left ventricular contractility were assessed in vivo and compared to the effects of adenosine and sodium nitroprusside, which exerts vasodilatory properties independently of cAMP. DESIGN AND SETTING: Prospective, randomized, cross-over in a university laboratory. SUBJECTS: Eleven pigs (25.9+/-2.8 kg, balanced anesthesia). INTERVENTIONS: Each animal was exposed to intravenous sodium nitroprusside, adenosine, and epoprostenol in randomized order. Iloprost was administered at the end due to its longer half-life. The dose was titrated to achieve a 25% reduction in diastolic aortic pressure. MEASUREMENTS AND RESULTS: Left ventricular contractility was assessed before, during, and after each intervention by determination of the endsystolic elastance with the conductance method. While there was no change in endsystolic elastance upon the infusion of adenosine and sodium nitroprusside; endsystolic elastance increased in the case of epoprostenol (57%) and iloprost (71%). CONCLUSIONS: Left ventricular contractility is increased in vivo by epoprostenol and iloprost but not by adenosine or sodium nitroprusside at equipotent hypotensive dose. A contribution of sympathetic reflex activation of cardiac nerves on the increase in left ventricular contractility cannot be completely ruled out. PMID- 12908097 TI - High variability and disomic segregation of microsatellites in the octoploid Fragaria virginiana Mill. (Rosaceae). AB - The objectives of the present study were to develop microsatellite markers for the wild strawberry, Fragaria virginiana, to evaluate segregation patterns of microsatellite alleles in this octoploid species, and assess genetic variability at microsatellite loci in a wild population. A genomic library was screened for microsatellite repeats and several PCR primers were designed and tested. We also tested the use of heterologous primers and found that F. virginiana primers amplified products in cultivated strawberry, Fragaria x ananassa Duch. and Fragaria chiloensis. Similarly, microsatellite loci developed from cultivated strawberry also successfully amplified F. virginiana loci. We investigated four microsatellite loci in detail, three developed from F. virginiana and one from cultivated strawberry. A survey of 100 individuals from a population of F. virginiana in Pennsylvania demonstrated high heterozygosities (H(e) or gene diversity ranged from 0.80 to 0.88 per locus) and allelic diversity (12-17 alleles per locus), but individual plants had no more than two alleles per locus. Segregation patterns in parents and progeny of two controlled crosses at these four loci were consistent with disomic Mendelian inheritance. Together these findings suggest that the genome of F. virginiana is "highly diploidized" and at least a subset of microsatellite loci can be treated as codominant, diploid markers. Significant heterozygote deficiencies were found at three of the four loci for hermaphroditic individuals but for only one locus among females in this gynodioecious species. PMID- 12908098 TI - New methods of dissecting the masticator space of the rhesus macaque ( Macaca mulatta). AB - The aim of our project was to develop a method to examine the deep masticator space of the rhesus macaque ( Macaca mulatta) in order to investigate the following particulars of the mandibular nerve: (1). its manner of entry into the masticator space, (2). its branching within spaces/canals in the spheno-temporal bony complex, and (3). the location of its principal divisions with reference to the lateral pterygoid plate and muscle heads. In order to access these structures it was necessary to develop novel lateral and medial approaches. These and the instruments used are described. The proximal branching of the mandibular nerve is described and contrasted with that of the human. The implications of nerve branching in humans with reference to (1). evolution, (2). electromyography and (3). anesthetic control are discussed. PMID- 12908095 TI - Osteoporosis after solid organ and bone marrow transplantation. AB - Organ transplantation has become increasingly common as a therapy for end-stage renal, liver, cardiac and pulmonary disease. The population of patients who have survived organ transplantation has grown dramatically over the last 2 decades. Although organ transplant recipients now benefit from greatly improved survival, long-term complications of organ transplantation, such as osteoporosis, adversely affect quality of life and must be addressed. In the early post-transplantation period, the effects of high dose glucocorticoids, combined with other immunosuppressive drugs such as cycosporine A and tacrolimus, cause rapid bone loss particularly at the spine and proximal femur. In this setting, fracture incidence rates as high as 25-65% have been reported. Treatment and prevention strategies must target this early post-transplant period, as well as the patient awaiting transplantation and the long-term transplant recipient. This review will discuss the clinical features of transplantation osteoporosis, the pathophysiology of post-transplantation bone loss and prevention and therapy of this unique bone disease. PMID- 12908099 TI - Association of genetic variation of the RIL gene, encoding a PDZ-LIM domain protein and localized in 5q31.1, with low bone mineral density in adult Japanese women. AB - Twin and family studies had shown that genetic factors are important determinants of bone mass. Multiple genes might be involved. One candidate gene, the reversion induced LIM gene ( RIL), is a PDZ and LIM-domain-containing protein and has been localized within the cytokine cluster of chromosome 5 (5q31.1). In a genetic study of 370 adult Japanese women, we investigated the correlation between radial bone mineral density (BMD) and a genetic variation (-3333T-->C) of the 5' flanking region of RIL gene. A significant association was identified between the RIL variation -3333T-->C and radial BMD ( r=0.15, P=0.003). The variation of the RIL locus may be an important determinant of osteoporosis. PMID- 12908100 TI - Extreme androgen resistance in a kindred with a novel insertion/deletion mutation in exon 5 of the androgen receptor gene. AB - Androgen insensitivy syndrome (AIS) is the most frequent cause of male pseudohermaphroditism resulting from target-organ resistance to androgen action. Individuals bearing the complete form of the disease (CAIS) present a female phenotype and a lack of pubic and axillary hair. In the present study, four 46,XY patients born in two generations from a kindred with a history of AIS were examined for genetic abnormalities in the androgen receptor gene (AR). All eight exons encoding the AR protein were individually amplified from genomic DNA followed by a mutation screening with single-strand conformation polymorphism analysis. Sequencing of the mutant AR revealed a novel insertion/deletion mutation in exon 5. A deletion of 7 bp is replaced by an insertion of 11 nucleotides, which represents a duplication of the adjacent downstream sequence. The mutation g.2640_2646delAGGATGC/2652_2662insTTCGCCCCTGA, results in a frameshift that introduces a premature termination signal TGA, nine codons downstream. Such a rearrangement predicts a truncation of the AR, thereby deleting a large portion of the ligand-binding domain (amino acid position 768 919). Furthermore, although this mutation breaks the translational reading frame starting from codon 760, examination of the complementary DNA suggested that it does not disturb mRNA splicing. These changes have been found in all the patients and appear to account for the observed absence of detectable androgen binding to the AR in cultured fibroblasts and for the CAIS phenotype in the kindred. This disorder represents the first insertion/deletion mutation of the AR that probably arose by a slipped-strand mispairing mechanism. PMID- 12908101 TI - Dynamic molecules: molecular dynamics for everyone. An internet-based access to molecular dynamic simulations: basic concepts. AB - Molecular dynamics is a rapidly developing field of science and has become an established tool for studying the dynamic behavior of biomolecules. Although several high quality programs for performing molecular dynamic simulations are freely available, only well-trained scientists are currently able to make use of the broad scientific potential that molecular dynamic simulations offer to gain insight into structural questions at an atomic level. The "Dynamic Molecules" approach is the first internet portal that provides an interactive access to set up, perform and analyze molecular dynamic simulations. It is completely based on standard web technologies and uses only publicly available software. The aim is to open molecular dynamics techniques to a broader range of users including undergraduate students, teachers and scientists outside the bioinformatics field. The time-limiting factors are the availability of free capacity on the computing server to run the simulations and the time required to transport the history file through the internet for the animation mode. The interactive access mode of the portal is acceptable for animations of molecules having up to about 500 atoms. PMID- 12908104 TI - Hypertension and multicystic dysplastic kidney. PMID- 12908102 TI - A novel extracellular subtilisin-like protease from the hyperthermophile Aeropyrum pernix K1: biochemical properties, cloning, and expression. AB - A novel extracellular serine protease designated Pernisine was purified to homogeneity and characterized from the archaeon Aeropyrum pernix K1. The molecular mass, estimated by SDS-PAGE analysis and by gel filtration chromatography, was about 34 kDa suggesting that the enzyme is monomeric. Pernisine was active in a broad range of pH (5.0-12.0) and temperature (60-120 degrees C) with maximal activity at 90 degrees C and between pH 8.0 and 9.0. In the presence of 1 mM CaCl(2) the activity, as a function of the temperature, reached a maximum at 90 degrees C but at 120 degrees C the enzyme retained almost 80% of its maximal activity. Activity inhibition studies suggest that the enzyme is a serine metalloprotease and biochemical data indicate that Pernisine is a subtilisin-like enzyme. The protease gene, identified from the sequenced genome of A. pernix, was amplified from total genomic DNA by PCR technique to construct the expression plasmid pGEX-Pernisine. The Pernisine, lacking the leader sequence, was expressed in Escherichia coli BL21 strain as a fusion protein with glutathione- S-transferase. The biochemical properties of the recombinant enzyme were found to be similar to those of the native enzyme. PMID- 12908103 TI - A pilot safety and efficacy study of OP-1 putty (rhBMP-7) as an adjunct to iliac crest autograft in posterolateral lumbar fusions. AB - The ability of bone morphogenetic proteins (BMPs) to induce bone formation has led to an increasing interest in the potential for their use in fusion surgery. The purpose of this multi-center clinical pilot study was to evaluate the safety of one such BMP-osteogenic protein 1, in the form of OP-1 putty-combined with autograft for intertransverse process fusion of the lumbar spine in patients with symptomatic spinal stenosis and degenerative spondylolisthesis following spinal decompression. Twelve patients with spinal stenosis and degenerative lumbar spondylolisthesis underwent laminectomy and partial or complete medial facetectomy as required for decompression of the neural elements followed by intertransverse process fusion by placing iliac crest autograft and OP-1 putty between the decorticated transverse processes. No instrumentation was used. Patients were followed clinically using the Oswestry scale and radiographically using static and dynamic radiographs to assess their fusion status. Independent and blinded radiologists assessed the films for the presence of bridging bone between the transverse processes and measured translation and angulation on dynamic films using digital calipers. In addition to bridging bone, less than or equal to 5 degrees of angular motion and less than or equal to 2 mm of translation were required to classify the patients as successfully fused, as per the definition of successful fusion provided by the FDA for use in clinical trials involving investigational devices to attain spinal fusion. Radiographic outcome was compared to a historical control (autograft alone fusion without instrumentation for the treatment of degenerative spondylolisthesis). All adverse events were recorded prospectively. The results showed 9 of the 12 patients (75%) obtained at least a 20% improvement in their preoperative Oswestry score, while 6 of 11 patients (55%) with radiographic follow-up achieved a solid fusion by the criteria used in this study. Bridging bone on the anteroposterior film was observed in 10 of the 11 patients (91%). No systemic toxicity, ectopic bone formation, recurrent stenosis or other adverse events related to the OP-1 putty implant were observed. A successful fusion was observed in slightly over half the patients in this study, using stringent criteria without adjunctive spinal instrumentation. This study did not demonstrate the superiority of OP-1 combined with autograft over an autograft alone historical control, in which the fusion rate was approximately 45%. The lack of adverse events related to the OP-1 putty implant in this study is in agreement with other studies supporting the safety of bone morphogenetic proteins in spinal surgery. PMID- 12908105 TI - Interactions between willows and insect herbivores under enhanced ultraviolet-B radiation. AB - We studied the effects of elevated ultraviolet-B radiation on interactions between insect herbivores and their host plants by exposing two species of phytochemically different willows, Salix myrsinifolia and S. phylicifolia, to a modulated increase in ultraviolet radiation in an outdoor experiment and monitoring the colonisation of insect herbivores on these willows. We examined the effect of increased ultraviolet-B (UV-B) radiation on (1) the quality of willow leaves, (2) the distribution and abundance of insect herbivores feeding on these willows, (3) the resulting amount of damage, and (4) the performance of insect larvae feeding on the exposed plant tissue. Six clones of each of the two willow species were grown in eight blocks for 12 weeks in the UV-B irradiation field. The clones were exposed to a constant 50% increase in UV-B radiation (simulating 20-25% ozone depletion), to a small increase in UV-A radiation or to ambient solar irradiation. We allowed colonisation on the willows by naturally occurring insects, but also introduced adults of a leaf beetle, Phratora vitellinae, a specialist herbivore on S. myrsinifolia. Increased UV-B radiation did not affect any of the measured indices of plant quality. However, numbers of P. vitellinae on S. myrsinifolia were higher in plants with UV-B treatment compared with UV-A and shade controls. In laboratory tests, growth of the second instar larva of P. vitellinae was not affected by UV-B treatment of S. myrsinifolia, but was retarded on UV-B treated leaves of S. phylicifolia. In addition, naturally occurring insect herbivores were more abundant on willows exposed to elevated UV-B radiation compared to those grown under control treatments. In spite of the increased abundance of insect herbivores, willows treated with elevated UV-B did not suffer more herbivore damage than willows exposed to ambient solar radiation (shade control). The observed effects of UV-B on herbivore abundance, feeding and growth varied significantly due to spatial variation in environment quality, as indicated by the UV-treatment x block interaction. The results suggest that (1) environmental variation modifies the effects of UV-B radiation on plant-insect interactions and (2) specialist herbivores might be more sensitive to chemical changes in their secondary host plants (S. phylicifolia) than to changes in their primary hosts (S. myrsinifolia). PMID- 12908106 TI - High-resolution genetic and physical mapping of the cauliflower high-beta carotene gene Or ( Orange). AB - Mutation in the cauliflower gene Or causes high levels of beta-carotene to accumulate in various tissues of the plant that are normally devoid of carotenoids. To decipher the molecular basis by which Or regulates carotenoid accumulation, we have undertaken the isolation of Or by a map-based cloning strategy. Two previously isolated, locus-specific, sequence-characterized amplified region (SCAR) markers that flank Or were employed for the analysis of a large segregating population consisting of 1632 F(2) individuals, and a high resolution genetic linkage map of the Or locus region was developed. To facilitate positional cloning, we constructed a cauliflower genomic library in a bacterial artificial chromosome (BAC) vector, using high molecular weight DNA from Or homozygotes. The BAC library comprises 60,288 clones with an average insert size of 110 kb, and represents an estimated 10-fold coverage of the genome. A BAC contig encompassing the Or locus was established by screening the library with a marker that is closely linked to Or and by identifying overlapping BAC clones by chromosome walking. Physical mapping delimited the Or locus to a 50 kb DNA fragment within a single BAC clone, which corresponds to a genetic interval of 0.3 cM. PMID- 12908107 TI - The evolutionary relationships of zebrafish genes tbx6, tbx16/spadetail and mga. AB - The T-box (Tbx) genes encode a family of transcription factors required for development of vertebrate embryos. In an attempt to discover human orthologues of the zebrafish (Danio rerio) tbx6 and tbx16/spadetail genes, we found that the most similar human (Homo sapiens) gene is the orthologue of mouse (Mus musculus) Mga (MAX gene associated). We have identified the zebrafish orthologue of Mga using analyses of sequence similarity and the orthologies of syntenic genes. Zebrafish mga maps close to dtk (developmental receptor tyrosine kinase), the orthologue of human TYRO3 (TYRO3 protein tyrosine kinase). Like its human and mouse orthologues, zebrafish mga lacks the three conserved introns within the T box coding sequences that are characteristic of the vertebrate T-box gene family. This suggests that these genes are derived from an ancient reverse transcription event. The human genome does not appear to possess orthologues of zebrafish tbx6 or tbx16/spadetail. PMID- 12908108 TI - Identification of the Drosophila melanogaster homolog of the human spastin gene. AB - The human SPG4 locus encodes the spastin gene, which is responsible for the most prevalent form of autosomal dominant hereditary spastic paraplegia (AD-HSP), a neurodegenerative disorder. Here we identify the predicted gene product CG5977 as the Drosophila homolog of the human spastin gene, with much higher sequence similarities than any other related AAA domain protein in the fly. Furthermore we report a new potential transmembrane domain in the N-terminus of the two homologous proteins. During embryogenesis, the expression pattern of Drosophila spastin becomes restricted primarily to the central nervous system, in contrast to the ubiquitous expression of the vertebrate spastin genes. Given this nervous system-specific expression, it will be important to determine if Drosophila spastin loss-of-function mutations also lead to neurodegeneration. PMID- 12908110 TI - Prevention of multiple pregnancies by restricting the number of transferred embryos: randomized control study. AB - INTRODUCTION: The high incidence of multiple pregnancies is a major concern in the treatment of infertility by in vitro fertilization (IVF). Risks and costs are associated with multiple pregnancies and it is important to find a way to decrease multiple pregnancies. The standard protocol for IVF includes the transfer of two or three embryos. METHODS: In this study, we compared the rate of single and multiple pregnancies between patients who received two and three embryos. One hundred and sixty-nine patients were randomly divided into two groups, with 106 cycles of IVF each. Group 1 received two embryos, and group 2 three. RESULTS: Group 1 had total 40 (37.8%) pregnancies, while group 2 had 29 (27.4%). The number of ongoing pregnancies was 33 (31.1%) in group 1 and 26 (24.5%) in group 2. The number of baby-take-home was 32 (30.1%) in group 1 and 26 (24.5%) in group 2. These results show that there was no significant difference between the two groups in both the total and ongoing pregnancies. On the other hand, the number of twin pregnancies was 6 (15.0%) in group 1 and 12 (41.4%) in group 2, showing a significant difference. Group 1 had no triplet pregnancies, while Group 2 had two. CONCLUSION: We conclude that the transfer of two embryos reduces the incidence of multiple pregnancies while maintaining an acceptable rate of pregnancy in IVF. PMID- 12908109 TI - Differential expression and localisation of WWP1, a Nedd4-like protein, in epithelia. AB - Ubiquitination of proteins such as ion transporters appears to be an important process in the regulation of their membrane expression. Recently, using two hybrid screening, we have selected a potential partner for the alpha-subunit of the amiloride-sensitive epithelial sodium channel (ENaC): the WWP1 protein, a ubiquitin ligase belonging to the Nedd4 family. To establish whether WWP1 is co expressed with ENaC, we employed in situ hybridisation, immunohistochemistry and Western blotting to determine the expression of WWP1 in various tissues and cell lines, including those known to express ENaC. As expected, WWP1 was expressed, like ENaC, in the bronchiolar epithelium. However it was also present in the proximal colon and the proximal part of the nephron (where ENaC is not expressed) and absent in the distal parts of the nephron (where ENaC is expressed abundantly). These results suggest that other channels or transport proteins, containing specific domains, such as PY motifs, could be the targets for regulation by WWP1. PMID- 12908113 TI - Fetal hydrocephalus--prenatal treatment. AB - PATIENTS AND METHODS: From January 1986 to January 2001, 85 cases of hydrocephalus were referred to the Pediatric Neurosurgery and Fetal Medicine Sections of the Sao Paulo Federal University and the Santa Joana Maternity Hospital in Sao Paulo, Brazil. Thirty-nine of these cases concerned fetuses ranging from 24 to 32 weeks' gestation, who underwent intrauterine treatment. The hydrocephalus etiologies consisted of: aqueduct stenosis (18 cases), intraventricular hemorrhage (8 cases), Dandy-Walker malformation (4 cases), Chiari type II (4 cases), and other diverse etiologies (5 cases). Twenty fetuses underwent repeated cephalocentesis: 18 received a ventriculo-amniotic shunt and in 1 case endoscopic third ventriculostomy was performed. The follow-up period varied between 1 and 14 years (median 5 years). Out of 39 patients, 26 have been considered as normal (IQ above 70), 6 have mild or moderate handicaps (IQ from 35 to 70), and 7 are severely handicapped (IQ below 35). After birth, 38 patients underwent ventriculoperitoneal shunting with low-pressure valves. CONCLUSION: In the absence of chromosomopathies, when the gestational age ranges between 24 and 32 weeks, progressive fetal hydrocephalus of a non-infectious origin can obtain benefit from intraventricular decompression performed during the fetal period. New methods of diagnosis and treatment should be developed, especially in those countries where interruption of gestation is not permitted by law, for the benefit of those families who, due to social or religious factors, do not accept abortion. PMID- 12908112 TI - Fetal brain tumors. AB - INTRODUCTION: Fetal intracranial tumors are rare and their evolution is usually poor. With routine use of antenatal ultrasound imaging, a larger number of such tumors have been diagnosed. METHODS: During the period from January 1992 to January 2002, 18 cases diagnosed as intracranial tumors in the fetal stage were treated in the Neurosurgery and Obstetrics Departments of the Federal University of Sao Paulo and in the Fetal Medicine Service of the Maternity Hospital Santa Joana. RESULTS: The tumors most frequently found were those of the choroid plexus and teratomas. Out of those 18 cases, 11 patients presented with fetal hydrocephalus, and 5 of them, who were less than 34 weeks old (gestational age), underwent repeated cephalocenteses while awaiting better conditions for delivery. Two fetuses died while in utero before any intervention had taken place and 16 underwent craniotomy after birth. Twelve of those patients are still alive, 9 of whom are considered normal. The 3 others are slightly or moderately retarded. PMID- 12908114 TI - Prenatal screening: invasive diagnostic approaches. AB - INTRODUCTION: Fetal invasive procedures provide ovular samples that are helpful in establishing diagnosis, etiology, and prognosis when ultrasonography and MRI show a central nervous system (CNS) anomaly or when the fetus is at high risk of such pathology. PROCEDURES: Invasive procedure techniques are amniocentesis, fetal blood sampling (FBS), and chorionic villous sampling (CVS). They provide material for fetal DNA, biochemical analysis, or identification of various infectious agents. COMPLICATIONS: The main complications are miscarriage and premature delivery. Counseling the parents about the risks and benefits of these procedures is therefore mandatory. DISCUSSION: Amniocentesis may be performed as early as 15 weeks' gestation and is the most widely used invasive technique. FBS is performed after 18 to 20 weeks of pregnancy and CVS may be carried out after 11 weeks. Indications for invasive techniques include DNA and cytogenetic analysis, diagnosis of neural tube defects, identification of infectious agents (toxoplasmosis, cytomegalovirus, rubella, and varicella), and etiology of intracerebral hemorrhage (fetal platelets and coagulation factors). PMID- 12908115 TI - Early decompressive craniotomy in children with severe traumatic brain injury. AB - INTRODUCTION: Decompressive craniectomy remains a controversial procedure in the treatment of raised intracranial pressure (ICP) associated with post-traumatic brain swelling. Although there are a number of studies in adults published in the literature on this topic, most commonly as a salvage procedure in the treatment of refractory raised ICP, there are few that investigate it primarily in children with head injuries. AIM: Our aim was to report the experience with decompressive craniotomy in children with severe traumatic brain injury (TBI) at the Red Cross Children's' hospital. METHODS: This study reports five patients in whom decompressive craniectomy or craniotomy with duraplasty was used as an early, aggressive treatment of raised ICP causing secondary acute neurological deterioration after head injury. The rationale was to save the patient from acute cerebral herniation and to prevent exposure to a prolonged course of intracranial hypertension. RESULTS: All patients benefited from the procedure, demonstrating control of ICP, radiological improvement and neurological recovery. Long-term follow-up was available, with outcome assessed at a minimum of 14 months after injury. DISCUSSION: The early approach to the use of decompressive craniotomy in the treatment of severe traumatic brain injury (TBI) with secondary deterioration due to raised ICP is emphasised. A favourable outcome was achieved in all of the cases presented. The potential benefit of decompressive craniectomy/craniotomy in the management of children with severe TBI is discussed. PMID- 12908116 TI - Acquired aqueductal stenosis in preterm infants: an indication for neuroendoscopic third ventriculostomy. AB - OBJECT: The object of this study is to demonstrate the delayed occurrence of aqueductal stenosis in preterm infants who have suffered from intraventricular hemorrhage (IVH) and to try to explain the mechanisms of this stenosis. METHOD: From January 1996 to June 2002, 1,046 premature infants were admitted to our institution. Thirty-six neonates suffered from grade 3 or 4 intraventricular hemorrhage (Papile grading), of whom 16 died. Twenty patients survived and a ventriculoperitoneal shunt was inserted in 7 infants. Four patients underwent a neuroendoscopic third ventriculostomy. Follow-up was carried out, twice a month during the first 2 months and subsequently twice a year. CONCLUSION: In 2 children NTV was an effective treatment for hydrocephalus with an average follow up of 29 months. The specific pattern concerning these patients is the long delay before obstructive hydrocephalus and the visualization of de novo obstruction with MRI. The biological explanation must be investigated. PMID- 12908117 TI - New orbitozygomatic approach by craniotomy. AB - INTRODUCTION: The orbitozygomatic extension technique described by Sekhar and Wright requires a temporal basal craniectomy that must be done in order to make osteotomy dissections. In the present work, a technique that includes the base and squamous temporal bone and a craniotomy instead of a craniectomy is shown. METHOD: The temporal and masseter muscles are dissected and separated from the temporal and zygomatic bones to facilitate passing them through the zygomatic arch in a caudal direction, which allows a frontotemporal basal craniotomy. Intracranially, a trepan that communicates to the glenoid cavity is done, which permits a temporal basal cut using the pneumatic drill saw. This is done in order to join with the orbital wall osteotomies, which were done in this approach with the craniotomy. PMID- 12908119 TI - The impact of touch-tone data entry on reports of HIV and STD risk behaviors in telephone interviews. AB - Respondents concerns about privacy can decrease reporting of HIV and STD risk behaviors in general population telephone surveys. The purpose of this paper is to describe the results of an experimental study evaluating whether one method for increasing privacy, touch-tone data entry (TTDE), is effective in increasing estimates of sexual behaviors from a population-based survey. We conducted a random-digit-dial telephone survey of adults in New Jersey (n = 405), with half the respondents using TTDE for answering sexual behavior questions. TTDE led to increased reports of same-sex sexual behavior, certain HIV and STD risk factors, and concern about one s risk for HIV and STD transmission. TTDE also narrowed the difference between men s and women s reports of the number of different sexual partners over the past 10 years. The feasibility and limitations of TTDE are discussed, as well as possible alternative interpretations that consider the impact of TTDE on the dynamics of the interaction between the respondent and the interviewer. PMID- 12908118 TI - Intramedullary spinal cord tumors in children. AB - INTRODUCTION: Pediatric intramedullary spinal cord tumors are rare and make up only a small percentage of all central nervous system neoplasms. DISCUSSION: These neoplasms are predominantly benign histologically, regardless of size, with a large percentage being astrocytomas or gangliogliomas. Ependymomas, which are common in adults, are relatively uncommon in children. Contemporary management of these tumors has generated much controversy. These neoplasms occur primarily in the cervical region, with pain or a motor deficit as the chief complaint. Surgery on these neoplasms can safely be performed using modern surgical adjuncts such as the ultrasonic aspirator, contact laser and neurophysiological monitoring. A radical resection of these tumors results in a good long-term outcome, since the majority are histologically benign. Adjuvant radiation therapy should only be administered for high-grade or malignant tumors. CONCLUSION: The majority of spinal cord tumors are benign; however, malignant tumors have a dismal outcome and surgery in these children should be limited to a conservative debulking. An attempt should be made for radical surgery in hope of avoiding radiation and chemotherapy for the benign tumors. Spinal deformity is a concern in these patients since 35% of children may require a stabilization procedure. PMID- 12908120 TI - Pluralistic ignorance and hooking up. AB - Hooking up--when two people agree to engage in sexual behavior for which there is no future commitment--has become popular on college campuses. In this study we examined the extent to which pluralistic ignorance affects hooking up. One hundred thirty-six female and 128 male college students answered questions regarding their own comfort and their perceived peers comfort in engaging in a variety of sexual behaviors while hooking up. We hypothesized and found that both women and men rated their peers as being more comfortable engaging in these behaviors than they rated themselves. Men expressed more comfort than did women in engaging in these behaviors, and both sexes overestimated the other gender s comfort with hooking up behaviors. Pluralistic ignorance appears to apply to hooking up on college campuses, and we explore some potential consequences of pluralistic ignorance in this context. PMID- 12908121 TI - Priming the sexual system: implicit versus explicit activation. AB - We investigated implicit versus explicit activation of the sexual system using a priming paradigm in which sexual slides were preceded by either sexual or neutral primes. In the first experiment, primes were made inaccessible to conscious cognitive elaboration. Identification of sexual targets was facilitated by sexual primes, indicating that sexual representations in memory can be activated automatically. In the second experiment, in which primes were presented at a conscious level, identification of sexual targets was decelerated by sexual primes. Primes elicited subjective sexual arousal in Experiment 2 only, demonstrating that the activation of subjective experience requires conscious cognitive elaboration. With the addition of a sexually specific physiological measure, to be constructed to measure initial genital responses, this paradigm may help elucidate activational mechanisms of sexual response. PMID- 12908122 TI - The lives and voices of highly sexual women. AB - American women who experience very strong and frequent sexual desire have often been either ignored or stigmatized. This exploratory study of 44 highly sexual women ages 20 to 82, which used a convenience sample and a semi-structured in depth interview method, allowed participants to describe their lives and how their sexuality has affected them. Highly sexual women reported that their lives have been strongly affected by their sexuality. First, the participants internally sensed demand for sexual excitement and satisfaction is too intense to be ignored. For some it was a major organizing principle of their time and energy. Second, their lives are lived in a society that often defines highly sexual women in pejorative ways. Women reported experiencing struggles and challenges in almost every area of their lives because of their sexuality, including feelings about themselves and their relationships with partners, female friends, and acquaintances. PMID- 12908123 TI - Casual sex among Australian schoolies. AB - This study of the casual sexual partnering of 570 male and 776 female Australian high school students on a schoolies week vacation expands on earlier research on factors that influence the sexual activity of vacationing youth. Over 60% of the men and nearly 40% of the women who engaged in sexual intercourse during schoolies week did so with a casual partner. We used logistic regression to test an expanded version of Triandis Theory of Interpersonal Behavior (TIB) in explaining casual sexual partnering. Situational experiences (similar to what others have called situational disinhibition), prior sexual experiences and intentions (similar to what others have called spillover) influenced casual sexual partnering. Different causal pathways were demonstrated for men and women. PMID- 12908124 TI - Heterosocial competence of rapists and child molesters: a meta-analysis. AB - A meta-analysis of 119 effect sizes from 14 papers examined the heterosocial competence of sex offenders. Rapists had significantly lower heterosocial competence than non-sex-offenders, but this effect was relatively small. Rapists in prison differed from non-sex-offenders who were not in prison; however, they did not differ from non-sex-offending prisoners. When unreliable measures were excluded from the analysis, nonincarcerated students and other volunteers who reported committing rape did not differ from nonincarcerated non-sex-offenders. The difference in heterosocial competence between child molesters and non-sex offenders was significantly larger than the difference between rapists and non sex-offenders. Also, child molesters had significantly less heterosocial competence than rapists. Findings clarify the relationship between heterosocial competence, rape, and child sexual abuse. Implications for the research and treatment of sex offenders are discussed. PMID- 12908125 TI - Sexual behavior among HIV-positive men who have sex with men: what's in a label? AB - Relatively little attention has been paid to the use and importance of labels used by men who have sex with men to describe insertive or receptive sexual behavior during intercourse. This study examines sexual self-labels, sexual behavior, HIV transmission risk, and psychological functioning among 205 HIV seropositive men who have sex with men. The majority of participants (88%) identified as a top, a bottom, or versatile. Tops were more likely to engage in insertive anal intercourse than bottoms, and bottoms were more likely to engage in receptive anal intercourse than tops, with versatiles reporting intermediate rates of both behaviors. Although the results suggest preliminary evidence regarding the predictive utility of self-labels, sexual behaviors of self-label groups were greatly overlapping. Differences were found among self label groups in gay self-identification, internalized homophobia, sexual sensation seeking, and anxiety. Results suggest an added value in assessing self labels in addition to asking about sexual behavior. PMID- 12908126 TI - Reasons for the adoption of celibacy among older men and women living with HIV/AIDS. AB - The reasons for becoming celibate following diagnosis with HIV/AIDS were examined using focused interviews with 63 infected older adults (ages 50-68). Forty-eight percent reported they were currently celibate or had been celibate following diagnosis with HIV/AIDS. Women reported celibacy (78%) more than men (36%). Although men and women reported some similar reasons for celibacy, most notably fear of infecting others and fear of reinfection, we also found gender differences in the reasons for celibacy. Additional reasons offered by women included loss of interest in sex, anger and distrust of men, and desire to focus on themselves rather than men. Other reasons offered by men included fear of rejection or stigmatization, difficulty with sexual performance, and negative body image. The prevalence of celibacy and the finding that many reasons for celibacy are related to fear, anger, and distrust suggests that older adults may have difficulty resuming healthy sexual relationships following diagnosis with HIV/AIDS. PMID- 12908127 TI - Precursors and pathways to adolescent prostitution in Taiwan. AB - This paper identifies the major precursors and pathways to prostitution for contemporary Taiwanese girls and the socioeconomic and cultural changes that have facilitated the shift in pathways over the past 10 years. Special attention is paid to the case of Taiwanese aborigines, who have particularly suffered systemic disruption due to prostitution. Analysis of case files and interviews provide quantitative and qualitative data that highlight the role of filial piety and the shift from indenture to runaway as the major pathway in Taiwan. Discussion includes relation of these findings to juvenile entry into prostitution in other countries, directions for future research, and policy recommendations for Taiwan's government. PMID- 12908128 TI - Ethnicity or race, area characteristics, and sexual partner choice among American adolescents. AB - The aim of this paper is to examine the association of individual demographic variables and area characteristics with the characteristics of sexual partners of American adolescents. Data for the study were drawn from the National Longitudinal Study of Adolescent Health. The data indicate that the community characteristics of ethnic or racial composition of the population and region were most strongly related to the ethnicity or race of the partner. Regional differences in the ethnicity or race of partners were significant even after controlling for the ethnic or racial composition of the community. Differences in partner s age, grade, neighborhood, and school or residence were not related to school and community characteristics. In addition, the individual characteristics of age, gender, and race or ethnicity were also related to ethnicity or race of partner. PMID- 12908132 TI - Chemotherapy and irradiation interaction. AB - The combination of chemotherapeutic drugs with radiation therapy has perhaps been one of the most promising strategies for current cancer radiotherapy practice, and has a strong biologic rationale. Concurrent chemoradiotherapy has been shown to improve survival and locoregional control of various cancers compared with radiotherapy alone. This article reviews the biologic rationale and principles fundamental to the use of chemoradiotherapy, and discusses mechanistic interactions between drugs and radiation, the knowledge of which is essential in developing optimal treatment strategies. Chemotherapeutic agents, particularly those shown to augment the radiation response of tumors, will be reviewed, and novel compounds and some of the exciting emerging strategies for chemoradiotherapy will be presented. PMID- 12908130 TI - Description, nomenclature, and mapping of a novel cerebello-renal syndrome with the molar tooth malformation. AB - Cerebello-oculo-renal syndromes (CORSs) and Joubert syndrome (JS) are clinically and genetically heterogeneous autosomal recessive syndromes that share a complex neuroradiological malformation resembling a molar tooth on brain axial images, a condition referred to as "molar tooth on imaging" (MTI) or the "molar tooth sign." The current literature on these syndromes is complex, with overlapping and incomplete phenotypes that complicate the selection of clinically homogeneous cases for genetic purposes. So far, only one locus (JBTS1 on 9q34) has been mapped, in two families with JS. Here, we describe a large consanguineous family with JS and nephronophthisis, representing a novel cerebello-renal phenotype. We have mapped this condition to the pericentromeric region of chromosome 11 and have named the locus "CORS2." The acronym "CORS" is proposed for all loci associated with JS, CORSs, and related phenotypes sharing the MTI, because this neuroradiological sign seems to be the unifying feature of these clinically heterogeneous syndromes. PMID- 12908133 TI - Combined modality treatment for central nervous system malignancies. AB - This review summarizes the current status and future prospects for combined modality treatment of primary and metastatic central nervous system malignancies. The laboratory and clinical basis for multimodality therapy, including surgery, ionizing radiation, and drug therapy, are outlined and critically reviewed. The central nervous system diseases discussed include: glioma (low and high grade), brain metastases, and primary central nervous system lymphoma. Collectively, these data suggest a shift favoring combined modality approaches in several of these diseases; however, the incremental gains are indeed modest. The individual practitioner must weigh these with the additional toxicities before making a therapeutic decision for a particular patient. The future direction of combined modality therapy in these diseases will likely revolve around the increased use of molecular diagnostics resulting in the application of targeted therapy. Clearly, such promising innovations must be delineated in the context of continued preclinical studies and controlled clinical trials. PMID- 12908134 TI - Combined modality management of breast cancer: development of predictive markers through proteomics. AB - The treatment of localized breast cancer often requires the use of surgery, radiation, and chemotherapy. The optimal sequencing of these treatments remains controversial. Although randomized studies have not found an improvement in survival with the use of neoadjuvant chemotherapy, a higher percentage of patients can undergo breast conservation. This clinical setting also allows us the unique opportunity to measure in vivo response to a select drug or drug combination. While there have been many retrospective studies looking at prognostic markers of response, there have been very few prospective studies developing markers that predict ultimate outcome from a specific drug or drug combination. This review discusses some of the newer technologies being used to develop predictive markers of therapeutic response. Proteomics is the study of the complete set of proteins expressed in a cell. It is a powerful tool to compare two different samples. Serial biopsies that compare the protein profile before and 24 hours after a cycle of chemotherapy may allow us to determine particular protein profiles that predict for ultimate clinical outcome. PMID- 12908135 TI - The role of concurrent chemotherapy and radiation in the management of patients with squamous cell carcinomas of the head and neck. AB - The management of patients diagnosed with squamous cell carcinomas of the head and neck has undergone a major paradigm shift during the past decade. The current application of systemic therapy and radiation has offered this patient population the opportunity for cure while simultaneously achieving organ preservation. Researchers have focused their efforts on optimizing radiotherapy treatment schedules, defining effective chemotherapy combinations, and investigating novel drug delivery procedures. This article will review the results of the early trials that identified the ability of sequential therapy to achieve organ preservation without sacrificing survival. These studies helped pave the way for the development of concurrent chemoradiation strategies. The results of multiple meta-analyses that have served as the basis for recent multi-institutional, randomized trials will be examined. At the same time, investigators have studied the role of altered fractionation radiotherapy schedules in conjunction with both systemic and intra-arterial chemotherapy. Researchers have long appreciated the differences in outcomes of head and neck tumors based on anatomic location. The results of the Intergroup trial that has helped to define the current national standard of care for this patient group will be reviewed. While most studies conducted to date have used platinum and 5-fluorouracil drug combinations, recent trials have begun to investigate the use of taxane-based regimens. The ability to now target cellular protein receptors has led investigators to focus their interest on the potential role for new biologic agents in combined modality treatment schemes. These treatment regimens hold tremendous promise for improving long-term outcomes while maintaining the quality of life for this patient population. PMID- 12908136 TI - Combined modality therapy for esophageal cancer. AB - Treatment approaches for esophageal cancer include primary treatment (surgical or nonsurgical) or adjuvant treatment (preoperative or postoperative). Primary treatments include surgery alone, radiation therapy alone, and radiation therapy plus chemotherapy (combined modality therapy). Adjuvant therapies include preoperative or postoperative radiation therapy, preoperative chemotherapy, and preoperative combined modality therapy. There is considerable controversy as to the ideal therapeutic approach. This review will examine the results of these approaches as well as combined modality therapy using novel regimens. PMID- 12908129 TI - Natural selection on the olfactory receptor gene family in humans and chimpanzees. AB - The olfactory receptor (OR) genes constitute the largest gene family in mammalian genomes. Humans have >1,000 OR genes, of which only approximately 40% have an intact coding region and are therefore putatively functional. In contrast, the fraction of intact OR genes in the genomes of the great apes is significantly greater (68%-72%), suggesting that selective pressures on the OR repertoire vary among these species. We have examined the evolutionary forces that shaped the OR gene family in humans and chimpanzees by resequencing 20 OR genes in 16 humans, 16 chimpanzees, and one orangutan. We compared the variation at the OR genes with that at intergenic regions. In both humans and chimpanzees, OR pseudogenes seem to evolve neutrally. In chimpanzees, patterns of variability are consistent with purifying selection acting on intact OR genes, whereas, in humans, there is suggestive evidence for positive selection acting on intact OR genes. These observations are likely due to differences in lifestyle, between humans and great apes, that have led to distinct sensory needs. PMID- 12908137 TI - Combined treatment for limited small cell lung cancer. AB - Radiation therapy is an important component for the management of limited small cell lung cancer (SCLC) in addition to systemic treatment. The role of surgery is limited for SCLC because of the nature of the disease, which often spreads to regional lymph nodes or spreads distantly. There have been major advances in the treatment of limited-stage SCLC during the past decade because of early application of thoracic radiation therapy (TRT) with concurrent chemotherapy, accelerated TRT, the application of prophylactic cranial irradiation for patients who achieved complete response to concurrent chemotherapy and TRT, and the improvement in supportive care such as discontinuation of tobacco smoking after the diagnosis of SCLC, nutritional support, pain control, granulocyte colony stimulating factor, erythropoietin, and adequate antibiotics. It is important to select patients with limited SCLC who are able to tolerate concurrent chemotherapy and accelerated TRT. PMID- 12908139 TI - Combined modality therapy in the treatment of gynecologic malignancies. AB - During the last several years, combined modality therapy has become increasingly important in the treatment of several malignancies. This trend holds true in the treatment of the gynecologic malignancies. In this review we examine the evolving role of combined modality therapy in the treatment of patients with cervical, vulvar, endometrial, and ovarian cancers. Definitive combined chemoradiation has become the new standard of care for patients with locally advanced cervical cancer. Combined chemoradiation provides for improvements in many clinical endpoints, including overall and disease-free survival. Chemoradiation has been used with some success in the treatment of patients with locally advanced vulvar carcinoma and has allowed some patients to avoid disfiguring surgery. Combined chemoradiation is being used in the treatment of patients with endometrial cancer who are found to be at high risk of recurrence after hysterectomy. In the setting of ovarian cancer, abdominopelvic radiation, either with or without chemotherapy, has shown some promise in the treatment of patients with minimal residual disease after resection. Combined modality therapy has played an increasingly important role in the treatment of gynecologic malignancies and its role continues to evolve and expand. PMID- 12908138 TI - Combined modality therapy for pancreatic cancer. AB - In spite of the high mortality in pancreatic cancer, significant progress is being made. This review discusses multimodality therapy for patients with pancreatic cancer. According to several phase II trials and Gastrointestinal Tumor Study Group results, improvements in locoregional control and survival may be achieved when chemotherapy is added to radiation for locally advanced pancreatic cancer. Radiosensitizing chemotherapy agents such as 5-fluorouracil, gemcitabine, paclitaxel, and cisplatin have shown promise. Selected patients with locally advanced disease have been downstaged with chemoradiation, facilitating surgical resection. For patients with resectable disease, the completion of Radiation Therapy Oncology Group study 97-04 represents a major achievement and brings gemcitabine into the forefront of adjuvant therapy. Neoadjuvant chemoradiation may eliminate the delay of initiating adjuvant treatment and spare unnecessary surgery for patients with rapid systemic progression. Molecular agents are being combined with chemoradiation in an attempt to delay or prevent systemic progression. PMID- 12908140 TI - Combined modality treatment for prostate cancer: role of chemotherapy. AB - Chemotherapy for prostate cancer currently has an established role in the treatment of hormone-refractory prostate cancer. There is strong evidence that combined modality treatment with androgen ablation in addition to radiotherapy provides a benefit above and beyond radiotherapy alone in patients with a poor prognosis, perhaps because of an effect of androgen ablation on subclinical distant disease. There are currently several clinical trials investigating whether the addition of chemotherapy with known efficacy in the hormone refractory setting can provide a survival advantage when used adjuvantly. The role of chemotherapy for patients with nonmetastatic, hormone-naive prostate cancer will be discussed. PMID- 12908141 TI - Combined modality therapy for rectal and colon cancer. AB - Combined modality therapy (CMT) with radiation therapy and chemotherapy plays an important role in the management of rectal cancer. Postoperatively, pelvic irradiation and 5-fluorouracil-based chemotherapy have been used to improve local control and survival for high-risk patients after local excision, as well as for patients undergoing abdominoperineal or low anterior resection. For patients treated preoperatively, CMT can also be used to facilitate sphincter-sparing surgery for distal rectal tumors. Preoperative CMT in conjunction with surgery and intraoperative radiation therapy has been useful for patients with locally advanced and recurrent rectal cancer as well. While chemotherapy has benefit for patients with stage III colon cancer above the peritoneal reflection, the role of CMT is less well defined. Retrospective reviews suggest improved local control and survival in subgroups of patients treated with postoperative radiation therapy and chemotherapy. Active areas of investigation in CMT for colorectal cancer include the integration of biologic predictors, radiation modulators, and novel systemic agents. PMID- 12908142 TI - The rationale and potential of combining novel biologic therapies with radiotherapy: focus on non-small cell lung cancer. AB - There are a number of novel, biologically targeted therapies, including anti epidermal growth factor receptor agents (eg, ZD1839 and C225 [cetuximab]), anti angiogenic agents (eg, vascular endothelial growth factor receptor inhibitors endostatin and angiostatin), and vascular targeting agents (eg, ZD6126), currently undergoing clinical development for a range of human cancers. These agents generally have milder toxicity profiles than conventional cytotoxic agents, and have the potential to be combined effectively with radiotherapy and other conventional modalities in the treatment of malignancies. In this review we consider the mechanistic rationale for combining biologically targeted agents with radiotherapy, present preclinical data to validate this hypothesis, and discuss early clinical investigations of these novel combination therapies, with an emphasis on ongoing trials in non-small cell lung cancer. PMID- 12908145 TI - [Clinicopathologic observations on chronic hepatitis C therapy with interferon alpha]. AB - INTRODUCTION: Chronic hepatitis C is a progressive liver disease. It may lead to cirrhosis within 15-20 years, on the grounds of which hepatocellular carcinoma may develop in around 4% of the cases. The entity can thus be regarded a premalignant condition. Based on the above, early diagnosis is of great importance, as is prompt treatment. AIMS: 1. to find correlations between the activity grade, stage of the disease and the various histologic characteristics, which are befitting representations of the clinical course for chronic hepatitis C, 2. to collect data on changes of nuclear proteins as markers of cell proliferations, 3. to analyse the changes in the expression of the components of the extracellular matrix, which play role in the pathogenesis of the fibrosis. RESULTS: Analysing the 106 biopsy specimens most frequently mild degree of inflammation was found. The majority of cases were in stage 1 (41.5%), however 8.69% were already in stage 4. Lymphoid aggregates/follicles and bile duct lesion were in correlation with the activity of the disease. A considerable decrease in the ALT level and in the histologic activity was induced by the 12 months interferon-alpha treatment. A positive correlation was found for the expression of the nuclear proteins used as proliferation markers in minimal and mild activity cases, while in moderate chronic hepatitis C a remarkable decrease was manifest in respect to cyclin-A activity. After 12 months IFN-alpha treatment the proliferating cell nuclear antigen activity increased, with a decrease in cyclin A positivity. CONCLUSIONS: Chronic hepatitis C cases with mild activity may also belong to stage 4, the need for liver biopsies should therefore be considered even besides the presence of near normal biochemical parameters in certain cases. The positive correlations between lymphoid follicles/aggregates, bile duct lesions and chronic hepatitis C activity refers to the fact that the above noted histologic differences may be evaluated as prognostic factors. The 12 months interferon-alfa treatment has a moderating effect on cell proliferation, as shown by the lowered cyclin-A expression and does have an effect on particular components playing role in the early phase of fibrogenesis. PMID- 12908146 TI - [Heat shock proteins as chaperones of the immune response. The optimal stress of life]. AB - The author reviews the complex role of heat-shock proteins during the immune reaction, especially the cellular effects of heat-shock proteins during recognition processes by innate immunity. The pathogenetic role of heat-shock proteins in two multifactorial diseases, inflammatory bowel disease and atherosclerosis, is summarized. A new hypothesis is presented dealing with the "immunodeficiency burden" as the aggregate effect of the genetic polymorphisms resulting in disease susceptibility. According to this mechanism the aggregate effect of polymorphisms resulting in failure of protective immunity determines the susceptibility for the disease in the given subject, when a specific environmental factor is present. PMID- 12908147 TI - [Changes in hematologic and hemostatic parameters after transjugular intrahepatic portosystemic shunt (TIPS) implantation]. AB - BACKGROUND: The pathogenesis of thrombocyte- and leukopenia associated with liver cirrhosis is far from being understood. Hypersplenism is considered to play a major role in this hematologic complication. The effect of transjugular intrahepatic portosystemic shunt (TIPS) implantation--a more recent technique in portal decompression--on platelet count is controversial in the literature. One of the main problems related to TIPS is the frequent occurrence of shunt malfunctions. There have been no reports on consistent clinical or biochemical parameters being able to predict the occlusive and rebleeding episodes after TIPS implantation. AIM AND METHOD: Platelet counts, white blood cell counts and different haemostatic data (prothrombin time, activated partial thromboplastin time and fibrinogen level) of the 24 patients undergoing TIPS placement were analyzed retrospectively prior to the procedure, after one month, after 3 months and 3 monthly thereafter for 18 months. RESULTS: The portal pressure gradient decreasing below the desired 12 mmHg after TIPS placement seems to be the only factor, which can result in moderate but significant increase in platelet counts. There was no significant alteration in white blood cell counts during the follow up period. The different haemostatic parameters scattered in a wide range, no real tendency was demonstrable. Patients in whom recurrent variceal bleeding occurred, the platelet count at the 3rd month was significantly lower compared to the basal platelet count. The decrease in the platelet count preceded shunt malfunction detected with color-Doppler or the appearance of the clinical symptoms. CONCLUSION: Monitoring platelet count may be of prognostic interest in the assessment of the shunt function and the risk of imminent variceal rebleedings during the follow-up period. PMID- 12908148 TI - [Histological and immunohistochemical structure of pulmonary tuberculotic granulomas in untreated cases and cases treated with antitubercular drugs]. AB - INTRODUCTION: The changes occurring in response to antituberculotic treatment and immune defence were studied in human tuberculotic granulomas. AIMS: To compare the possibilities of detection of Mycobacterium tuberculosis with the Ziehl Neelsen staining technique and with an immunohistochemical method, and to assess the roles of lymphocytes and heat-shock protein 70. METHOD: 40 patients who had undergone lung resection (the postoperative histology confirmed tuberculosis) were divided into two equal groups, on the basis of whether they had received antituberculotic treatment preoperatively (group I) or not (group II). Customary histology was used to determine the Langhans cells, epitheloid cells and lymphocytes, and an immunohistochemical method was then applied to examine the heat-shock protein 70 production of these cells and the normal lung. The lymphocytes were divided into CD4+ T-helper, CD8+ T-cytotoxic and CD20+ B cells by means of immune examinations. M. tuberculosis was demonstrated by an immunohistochemical method, with antibody against the wall protein. RESULTS: Heat shock protein 70 was produced by 17.6% of the Langhans cells and 94.4% of the epitheloid cells in group I, and by 100% of both cell types in group II. The bacterium could be detected in 40% of the total number of cases with acid-fast staining, and in 85% by immunohistochemistry. There was no significant difference in the qualitative distribution of the lymphocytes in the granulomas in groups I and II. The heat-shock protein 70 levels of the tuberculotic granuloma and the normal lung were significantly higher in group II. CONCLUSIONS: The production of heat-shock protein 70 is more enhanced in untreated tuberculotic cases. On the basis of their heat-shock protein 70 production, the authors assume that a majority of the Langhans cells have a resting protective function in medically treated cases. Independently of the stage of the infection and of the use or not of antituberculotic treatment, the number of lymphocytes participating in the immune defence is constant. By means of immunohistochemical examination of the wall protein of M. tuberculosis, the presence of the tuberculotic disease can be demonstrated with high reliability. PMID- 12908149 TI - [Primary antiphospholipid syndrome associated with HELLP syndrome in pregnancy]. AB - Authors report the case of a pregnant woman with antiphospholipid syndrome associated with HELLP syndrome. Pregnancy was terminated because of severe preeclampsia. However the patient's condition worsened. New symptoms of antiphospholipid syndrome developed (deep vein thrombosis, ischaemic optic lesion) and ARDS, therefore respiratory therapy was introduced. Plasmaphereses were performed concomitantly with high-dose intravenous immunoglobulin, glucocorticoid, cyclophosphamide and anticoagulant therapy. She responded to the therapy well, and eventually recovered completely. There are only a few case reports about pregnancies complicated with the association of antiphospholipid syndrome and HELLP syndrome. To authors' best knowledge, this is the first Hungarian report about this rare and curious association and it's successful treatment. PMID- 12908150 TI - Institutional politics impede research. PMID- 12908151 TI - The changing CT environment. PMID- 12908152 TI - Sonographic assessment of fetal skeletal dysplasias. PMID- 12908153 TI - Molecular imaging of cancer: the basics. PMID- 12908154 TI - Total teleradiology. PMID- 12908155 TI - Portfolios and critical thinking. PMID- 12908156 TI - CT-gated cardiac scoring. PMID- 12908157 TI - Deceptive debris. PMID- 12908158 TI - The value of nonclinical staff. PMID- 12908159 TI - Patient page. Stereotactic breast biopsy. PMID- 12908160 TI - 2020 healthcare management in Canada: a new model home next door. AB - The Commission on the Future of Health Care in Canada asked whether Medicare is sustainable in its present form. Well, Medicare is not sustainable for at least six reasons. Given a long list of factors, such as Canada's changing dependency ratio, the phenomenon of diminishing returns from increased taxation, competing provincial expenditure needs, low labour and technological productivity in government-funded healthcare, the expectations held by baby boomers, and the evolving value sets of Canadians--Medicare will impoverish Canada within the next couple of decades if not seriously recast. As distasteful as parallel private pay, private-choice healthcare may be to some policy makers and providers who grew up in the 1960s, the reality of the 2020s will dictate its necessity as a pragmatic solution to a systemic problem. PMID- 12908161 TI - If the pillars are shaky, does faith in Medicare crumble? AB - The objectives of the study described in this article were to determine whether the faith of physicians in the Canadian system of health insurance depends on their assessment of quality and access to health services and whether their assessments of quality and access to health services affect their support of out of-pocket and other methods of financing healthcare. To this end, a mail survey of 600 physicians in British Columbia and 240 physicians in Saskatchewan was conducted. The sample was stratified to ensure equal representation from urban and non-urban areas and between female and male specialists and family practitioners. Our conclusions indicate that physicians seem to be open minded in their views on financing. Their overriding concerns are to ensure the provision of sufficient resources to the healthcare system and to maintain full coverage of the population. PMID- 12908162 TI - Day medicine: an urgent internal medicine clinic and medical procedures suite. AB - During Saskatchewan's healthcare reform of the 1990s, the number of acute hospital beds in Saskatoon District Health was cut in half. The emergency room and outpatient facilities were not able to accommodate an increasing number of patients needing urgent assessments and medical procedures. In this article, we describe the development, implementation and utilization of a day medicine program. This description may be useful to others planning healthcare delivery to medical patients, especially in a setting of resource constraint. PMID- 12908163 TI - Devolving healthcare delivery to regional health authorities: is health technology assessment prepared to follow? AB - Since the establishment of health technology assessment units in the latter 1980s, Canada has witnessed an unprecedented transformation of the governance, management and service delivery of its healthcare system. In Alberta, this transformation culminated in the establishment of regional health authorities that provide integrated healthcare to Albertans. With the shift of responsibility for healthcare delivery from the provincial to the regional level, the Alberta Heritage Foundation for Medical Research HTA unit recognized that for health technology assessment to continue to be relevant, it must follow this change. Four steps were taken to refocus the unit's scope: a thorough analysis of the healthcare environment; face-to-face interviews with the chief executive officers of the regions; the development of a framework for HTA in the regions; and the organization of a conference on evidence-based decision making. These steps were helpful in bringing HTA to the attention of regional decision makers. A formal, analytical assessment of the regional healthcare environment, provision of general information (through the framework and conference) and individual information (through face-to-face interviews) enabled a proactive engagement with regions. However, to meet the demands and needs of a population that expects comprehensive coverage that delivers "state of the art" diagnostics and treatments, the efficacy and effectiveness of interventions can sometimes be of subordinate importance. PMID- 12908164 TI - A test of a centre's vision. AB - The Thames Valley Children's Centre, a regional Children's Rehabilitation Centre, was determined to assess the extent to which its Vision ("Our Clients at Their Best") was being achieved. With a sample of 50 former clients, and utilizing a specially constructed survey tool based on the important life outcomes literature, they found that they were attaining their vision for a significant proportion of the participants. They also learned about the differing expectations of the role which the Centre should play. PMID- 12908165 TI - Going provincial: preceptoring and mentoring revisited in the health sciences. Part I. AB - This article describes the Preceptor & Mentor Initiative for Health Sciences in BC, a provincial strategy to facilitate preceptoring and mentoring. A web site was developed with a comprehensive preceptor/mentor framework and resources, based on best practices and applicable to all health sciences. Informal feedback indicates this Initiative is effective in supporting health sciences preceptoring and mentoring in BC. A formal evaluation will be conducted by March 2003 and reported in a future FORUM article. PMID- 12908166 TI - Issues, musings and trends IT & EHR governance: who should the leaders be? PMID- 12908167 TI - [Epidemiology of functional disorders of the kidneys]. AB - The new classification K/DOQI of chronic renal disease differentiates five stages according to the grade of glomerular filtration. In the fifth, most severe grade (= renal failure) dialyzation/transplantation treatment is indicated. For dialyzation/transplantation activities in developed countries relatively accurate epidemiological data exist (national and supranational registers of dialyzation and transplantation programmes). The number of patients increases year by year, in particular in the more advanced age groups. The predominating diagnosis is renal failure is diabetes. Although treatment by one of the methods substituting renal function involves less than 0.1% of the population the economic costs amount to as much as 6 = of the total expenditure on health services (Medicare data, USA). On the other hand the prevalence of chronic renal disease in less advanced stages is not exactly known. It is estimated that chronic kidney disease may be present in some 10% of the population. If the disease were detected in time and the patient was given professional care which can retard the disease, it would be not only of medical but also of socio-economic benefit. PMID- 12908168 TI - [Epidemiology of chronic kidney failure]. PMID- 12908169 TI - [Progression of chronic renal insufficiency and its prevention using angiotensin converting enzyme inhibitors and angiotensin antagonists]. AB - Progression of chronic nephropathies can be significantly retarded by optimal control of blood pressure. To achieve target blood pressure combined treatment is usually necessary in patients with chronic nephropathies. Drugs interfere with the renin-angiotensin-aldosterone system (angiotensin converting enzyme inhibitors and angiotensin antagonists) have as compared with other antihypertensive agents with a comparable blood pressure control a more marked renoprotective effect and are renoprotective also in normotensive microalbuminuric patients with diabetic nephropathy and in proteinuric patients with non-diabetic nephropathies. Treatment with angiotensin converting enzyme inhibitors and angiotensin antagonists should be started as soon as possible and should be lifelong. PMID- 12908170 TI - [Nephrogenic metabolic acidosis]. AB - Metabolic acidosis is a major risk factor of kidney disease progression as a consequence of impaired H+ urinary excretion by the decreased kidney NH3 synthesis. Two key enzymes participate: a) Phosphate-dependent glutaminase under the genomic control of metabolic acidosis and b) Phosphate independent glutaminase localized on proximal tubule microvili under the nongenomic control. Two types of kidney disease metabolic acidoses are dominant: a) Hyperchloremic metabolic acidosis usually on the basis of hereditary or toxic alterations, isolated or as a part of Fanconi syndrome. b) Hyperphosphatemic metabolic acidosis of renal insufficiency. Metabolic acidosis shares serious consequences: metabolic acidosis increases protein catabolism of amino acids, inhibits proteosynthesis (albumin!), accelerates renal osteodystrophy development, modulates calcidiol and parathormone plasma levels and evokes insulin resistance. The present therapy requires full correction of metabolic acidosis! PMID- 12908171 TI - [Metabolic and nutrition care in patients with chronic renal insufficiency and chronic kidney failure]. AB - Metabolic and nutritional care implies procedures which involve normalization or improvement of metabolic deviations in chronic renal insufficiency and failure by dietary and medicamentous means. The therapeutic procedure not only improves some metabolic disorders associated with a decline of the excretory and metabolic endocrinological renal function but can have a positive impact also on progression of renal insufficiency. Conservative treatment thus involves low protein diets, modification of electrolyte and water intake, adjustment of the acid-base balance, Ca, P metabolism, haemogram, hypertension, proteinuria and hyperlipidaemia. In non-diabetic subjects it is sufficient to achieve a glomerular filtration of 0.2 ml/sec. and serum creatinine of 500-600 mumol/l. PMID- 12908172 TI - [Anemia in chronic renal insufficiency and chronic failure]. AB - Complicated metabolic changes in chronic renal insufficiency interfere with the formation and survival of erythrocytes at many levels. Anaemia which thus develops influences in a significant way not only the quality of life but also the prognosis of nephrological patients. The confirmed relationship between cardiovascular morbidity and mortality must be the starting point for early diagnosis and treatment of this anaemia. Administration of recombinant human erythropoietin in the early stage of renal insufficiency can moreover probably retard its progression. European guidelines for the treatment of anaemia give clear instructions how to proceed in the treatment of anaemia. The results of current, so far not completed studies, may modify in future the demands on target haemoglobin values and may elucidate how normalization of the red haemogram can influence the survival of patients with impaired renal function. PMID- 12908173 TI - [Cardiovascular complications in patients with chronic renal insufficiency and chronic kidney failure]. AB - Patients with chronic renal failure have, as compared with age-matched controls with normal renal function, a markedly higher cardiovascular mortality. The reason is probably accelerated atherosclerosis and left ventricular hypertrophy as a result of accumulation of "classical" cardiovascular risk factors and the presence of some risk factors relatively specific for "uraemia" (e.g. anaemia, hyperhydratation, dyslipidaemia). It is assumed that the reversibility of left ventricular hypertrophy is limited in chronic renal failure due to more marked myocardial fibrosis ("uraemic cardiomyopathy"). Its regression can be achieved by treatment of hypertension with inhibitors of angiotensin converting enzyme with a positive effect on cardiovascular mortality. Regression of left ventricular hypertrophy occurs also in some patients after renal transplantation. Treatment of anaemia reduces the risk of progressive left ventricular dilatation. The cardiovascular risk increases probably already a relatively slight decline of glomerular filtration which need not lead to a significant rise of serum creatinine. The cardiovascular risk obviously increases further with progression of chronic renal insufficiency. Patients with a reduced renal function and chronic renal insufficiency have lower target blood pressure and should have also lower target values e.g. of serum cholesterol. Therapeutic procedures in these patients should not be focused only on a slower progression of chronic renal insufficiency but also on reduction of their high cardiovascular risk. PMID- 12908174 TI - [Hypertension and chronic renal insufficiency--chronic kidney failure]. AB - The prevalence of hypertension in patients with chronic renal insufficiency is high. In the stage of renal insufficiency it is 60% and in conservatively terminal renal failure it is as high as 90%. After the initiation of dialyzation treatment it declines temporarily, it is higher during chronic haemodialysis (50 80%) and lower in continuous ambulatory peritoneal dialysis (30%). After transplantation it is recorded in 70-80% recipients of a renal graft. Among the causes of renal hypertension in subjects with conservatively treated chronic renal insufficiency at present secondary renal impairment is increasing--in type 2 diabetes and also renal vascular affection due to atherosclerotic changes and essential arterial hypertension. Approximately 30% of patients where chronic dialyzation treatment is started, come "from the street". In the pathogenesis of renal hypertension sodium retention is involved with volume expansion and an impaired ratio of the formation of vasoactive (vasopressor and vasodepressor) substances. In chronic renal failure the volume component of hypertension predominates markedly. The causes of the development of hypertension after renal transplantation are multifactorial and are most closely associated with immunosuppressive treatment and graft rejection. Pharmacological treatment of renal hypertension prefers inhibitors of the angiotensin converting enzyme (possibly angiotensin II antagonists) because of their concurrent renoprotective action. In the stage of renal insufficiency they call for reduced doses and combination with other antihypertensive agents. The objective of treatment is to achieve a blood pressure < 130/80 mm Hg. In chronic dialyzation treatment the main therapeutic provision in hypertension is adjustment of the volume of extracellular fluid by regime provisions and effective haemoelimination treatment. Calcium blockers are useful in particular in the treatment of hypertension in haemodialyzed subjects and in hypertension after renal transplantation. PMID- 12908175 TI - [Diabetes mellitus and chronic renal insufficiency]. AB - In the nineties of the 20th century diabetic nephropathy has become the leading cause of regular dialysis treatment (RDT) in developed countries. In particular type 2 diabetics are involved. A similar trend can be observed also in the Czech Republic which holds in this respect the first place among countries of the former eastern block (33% patients with RDT) suffer from diabetes. The cause of the increase of patients with diabetic nephropathy and renal failure caused by diabetes is not only the rising prevalence and incidence of type 2 diabetes in the population but in particular the better care provided to patients with type 2 diabetes which enables them to survive macro- and microvascular complications incl. diabetic nephropathy. It is estimated that diabetic nephropathy affects 4 8% patients attending diabetic clinics. With regard to the increasing number of diabetics in RDT, moreover associated with their high polymorbidity, this is a serious medical and economic problem. The main factors which influence in the diabetic patients the risk of development of diabetic nephropathy are long-term control of glycaemia, genetic (ethnic) factors, age and sex. The decisive factor influencing in patients with diabetic nephropathy the progression of chronic renal insufficiency is control of the blood pressure. Including diabetics in RDT is not associated only with medical problems but also with socio-economic issues. The quality of life of diabetics is much lower and the survival of diabetics treated within the framework of RDT is still almost half as compared with the survival of non-diabetic patients. Decision on the selection of the dialysis method is not easy. Medical differences are well defined but should not be considered absolute. It is important to consider also which method is preferred by the patient. Optimally the decision is taken during the period of dispensarization. Both dialyzation methods have comparable results and survival although for diabetics under 50 years of age a more favourable prognosis of peritoneal dialysis is reported. However, the risk of "failure of the method" is in general higher in peritoneal dialysis. PMID- 12908176 TI - [Renal bone disease]. AB - Renal bone disease is a serious complication associated with chronic renal failure. The pathogenetic mechanisms are very complicated. The disorder develops as a result of hypophosphataemia, hypocalcaemia and calcitrol deficiency already during the period when renal functions decline below 50%. Formerly the form with an excessive bone turnover predominated, nowadays we encounter ever more frequently so-called a dynamic bone disease. A serious manifestation are extraosseous calcifications. In treatment phosphate binding substances in the gastrointestinal tract are involved (along with other provisions, correcting hypophosphataemia), supplementation of calcium in case of hypocalcaemia correction of metabolic acidosis and administration of the active vitamin D metabolite (continuously as supplementation in deficient endogenous production, in a pulsatile pattern with the aim to suppress the activity of parathyroid bodies). In case of "resistant" hyperparathyroidism surgery is indicated (parathyroidectomy). Treatment of the dynamic form is not known, prevention of suppression of excessive parathyroid activity is important. New trends in the treatment of renal bone disease are non-calcium phosphate binding substances in the gastrointestinal tract, vitamin D analogues (with a lower hypercalcaemic potential) and calcium mimetics. PMID- 12908177 TI - [Acute states in nephrology. Overview of problems]. AB - Acute conditions in nephrology are situations when treatment of the diagnosis and immediate therapeutic procedures to prevent damage of the kidneys and urogenital tract are urgent. Most important are conditions associated with oligoanuria or polyuria, vascular diseases with an acute course, acute obstructions of the urinary pathways and acute conditions in dialyzed and transplanted patients. An important group is also toxoallergic damage caused by drugs. PMID- 12908178 TI - [Hemodialysis in the treatment of chronic kidney failure. Present status]. AB - In advanced countries haemodialysis treatment is available to all patients with chronic renal failure who need it. At present nephrologists must resolve the problem when it is possible to with-hold long-term haemodialysis treatment, or withdraw it because it no longer leads to prolongation of a good quality life. The results of long-term dialysis treatment depend on the correct timing of its initiation and the quality of nephrological care provided already a long time before the development of renal failure. The morbidity, mortality and quality of life of the patients are influenced in a fundamental way by the quality of provided haemodialysis. An important factor is the dose of dialysis treatment evaluated according to index Kt/Vurea reflecting the urea elimination and obviously also the elimination of other low molecular weight substances. Although prospective controlled trials did not prove so far a favourable effect of haemodialysis membranes permeable for larger molecules ("high-flux" membranes) on the patients' fate, the possible toxic effect of so-called middle molecule substances and peptides with a low molecular weight is assumed. Data suggesting improvement of the quality of life of patients having daily haemodialyses call for further investigations. A still unresolved problem of contemporary haemodialysis systems remains inadequate biocompatibility which leads to reactions associated with possible acute and long-term damage of dialyzed patients. PMID- 12908179 TI - [Pitfalls in immunosuppressive therapy]. PMID- 12908180 TI - [Angiotensin receptor blocker often a valuable alternative to ACE inhibitors]. PMID- 12908181 TI - [Cannabis and schizophrenia]. PMID- 12908182 TI - [Should obesity be attacked by tax for overweight and censure?]. PMID- 12908183 TI - [Effects of ecstasy on the body]. PMID- 12908184 TI - [Muscle infarction--a rare complication of diabetes]. PMID- 12908185 TI - [Assessment of continuing medical education in pediatrics]. PMID- 12908186 TI - [Girl with asthma suffering from nicotine craving and feeling dizzy]. PMID- 12908187 TI - [Is autopsy a relevant procedure in modern times?]. PMID- 12908188 TI - [Is death biologically inevitable?]. PMID- 12908189 TI - [The roles of autopsies in the development of medicine]. PMID- 12908190 TI - [Autopsy in forensic medicine and the protection of law]. PMID- 12908191 TI - [The role of autopsy in diagnostics and clinical research]. PMID- 12908192 TI - [On death and body concepts in different religions]. PMID- 12908193 TI - Nursing theories: relic or stepping stone? PMID- 12908194 TI - Incorporating culture and gender into multiple intervention programs. PMID- 12908195 TI - "Like, what am I supposed to do?": adolescent girls' health concerns in their dating relationships. AB - This ethnographic study explored the health-related concerns, within dating relationships, of 40 female adolescents aged 15 and 16. The results reveal a complex interaction of male/female relational dynamics and socialization processes in these relationships. To avoid behaviours risky to their health, participants had to negotiate power relationships with partners and peers; yet, paradoxically, any increase in their power could increase the threat of violent confrontation, loss of power, and further health compromises. The girls' desire to have a dating partner outweighed their desire to avoid health threats such as substance abuse and violence. This dynamic can be understood in terms of patriarchal power imbalances. These findings point to the gravity of adolescent girls' health issues and the need for interventions that challenge mainstream culture and that foster empowerment among female adolescents. PMID- 12908196 TI - Stress disorders and gender: implications for theory and research. AB - Numerous epidemiological studies report increased prevalence rates for women as compared to men for stress-related disorders such as acute stress disorder, post traumatic stress disorder, and major depressive disorder. Stress disorders disrupt work and home life and pose a high risk for suicide. Multiple factors contribute to the increased vulnerability in women. Physiological differences account for some of the differential. Other factors that make a significant contribution to the overall risk for health problems in response to stressors or trauma include the nature and meaning of the trauma, accessibility of resources, and restrictive diagnostic categories. Increasing our knowledge of the individual impact of each factor as well as the interactions among the factors is central to understanding the development of stress disorders. Comprehensive sex- and gender sensitive middle-range theory, which explores the role of key factors identified in qualitative and quantitative research, is required. The authors discuss structural equation modelling as one method of theory testing. PMID- 12908197 TI - Peer sexual harassment in adolescence: the function of gender. AB - The author describes the sexual harassment experiences of 565 male and female students in Grades 9 through 11 and discusses the influence of the social construction of gender on those experiences. The sample was drawn from 12 schools in 2 Canadian provinces. The study employed a retrospective correlational survey design, using self-report questionnaires administered in class. The results indicate that sexual harassment is pervasive among both male and female adolescents but is particularly problematic for girls due to their qualitatively different harassment experiences. Both girls and boys reported gender harassment to be, in general, more upsetting than unwanted sexual advances. Girls employed a greater variety of coping strategies than boys; however, for both sexes passive coping strategies were predominant. The author offers recommendations related to school health programming and discusses the role of the school nurse. PMID- 12908198 TI - Mental health research and cultural dominance: the social construction of knowledge for international development. AB - This institutional ethnographic work uses the first author's experience as an international development worker, educator, and community mental health nurse in West Africa to illustrate how official research and policy on mental health services reflect Western academic, corporate, economic, and cultural dominance. Focusing on a critical textual analysis of a survey intended to support funding applications to international aid/lending agencies, the authors show how official processes privilege Western policies/research approaches and subordinate local perspectives. If nurses, researchers, and policy-makers are to be effective in carrying out development work in Africa, they must learn to appreciate the subtle exertion of dominance inherent in Western approaches. The authors propose that understanding local knowledge be foregrounded rather than backgrounded to the complex global interpretive frames for international research and international development policy. PMID- 12908199 TI - Daughters of cardiac patients: the process of caregiving. AB - This prospective, exploratory study examined the caregiving role that adult daughters play when a parent is hospitalized for a cardiac condition. Specifically, the study addressed the development and evolution of the caregiving role during the hospital stay and in the post-discharge period. Data were collected from 9 daughters using participant observation and unstructured and semi-structured interviews. Daughters' caregiving was characterized by a non linear process of "knowing" that included knowledge seeking, consolidation of knowledge, and acting on the knowledge. In a secondary process of self-assessment concerning their knowledge, the participants moved back and forth among the 3 dimensions. In addition, their level of involvement was defined by 4 different caregiving styles, based on the family's caregiving culture. Nurses might collaborate more effectively with women caring for a parent by determining their position in the "knowing" process and tailoring the intervention accordingly. Nurses should also be sensitive to families' opinions and traditions regarding caregiving. PMID- 12908201 TI - Translating and adapting measurement instruments for cross-linguistic and cross cultural research: a guide for practitioners. PMID- 12908200 TI - The home environment of Metis, First Nations, and Caucasian adolescent mothers: an examination of quality and influences. AB - This longitudinal exploratory study compared maternal psychosocial, situational, and home-environment characteristics at 4 weeks and at 12-18 months postnatal for a convenience sample of 71 Metis, First Nations, and Caucasian adolescent mothers. The combined group of Metis/First Nations mothers had significantly higher infant-care emotionality scores than the Caucasian mothers at 4 weeks. The Caucasian mothers scored considerably higher on quality of the home environment; a refined multiple regression model containing infant-care emotionality, education level of the infant's maternal grandmother, ethnicity, and enacted social support explained 49% of the variance, with significant influences being infant-care emotionality and grandmother's education level. PMID- 12908202 TI - Maternal-child health care in Aboriginal communities. PMID- 12908203 TI - Consequences of the Amsterdam Declaration--a rights revolution in Europe? PMID- 12908204 TI - Legal issues in quality of care oversight in the United States: recent developments. PMID- 12908205 TI - Human genomic databases: a global public good? PMID- 12908207 TI - Private insurance for all in the Dutch health care system? PMID- 12908206 TI - External peer review by medical specialist (visitatie) in a legal perspective. PMID- 12908208 TI - Laws and practices relating to euthanasia and assisted suicide in 34 countries of the Council of Europe and the USA. PMID- 12908209 TI - Give your reinsurance policy a thorough check up. PMID- 12908210 TI - Review of approaches to financing children's behavioral health favors carve-outs. PMID- 12908211 TI - Cap for injectables not for the faint of heart. PMID- 12908212 TI - Check your cap rates against 'allowable' reimbursement rates. PMID- 12908213 TI - Medicare moves closer to capitated disease management. PMID- 12908214 TI - The economic effect of hypoglycemia in a health plan. AB - This study was undertaken to determine the incidence and economic cost of hypoglycemia in patients with diabetes taking insulin in a large mid-western health plan. Analysis of claims data identified 7,659 members with a diagnostic code for either type 1 or type 2 diabetes during a seven-year interval from 1992 to 1998. A total of 2,118 were prescribed some type of insulin regimen. During the six years of follow-up, more than 16% of insulin-treated patients experienced hypoglycemia profound enough to warrant medical attention. The mean cost per episode was $1,186 (range, $181-$4,924) or $7.04 per patient per month. Although the financial burden is smaller than that of late complications of diabetes, hypoglycemia associated with insulin therapy is associated with significant costs to the health care system. PMID- 12908215 TI - A comparison of costs for four oral antidiabetic regimens within a managed care population. AB - A retrospective database analysis compared costs among patients with type 2 diabetes receiving four antidiabetic regimens: (1) repaglinide monotherapy, (2) metformin monotherapy, (3) repaglinide and metformin in combination, or (4) metformin and glyburide in combination. Pharmacy, medical, and total costs were measured for each cohort over a nine-month period. Although not statistically significant, total adjusted costs were lowest for the repaglinide-metformin combination ($8,924), followed by metformin monotherapy ($9,448), metformin and glyburide ($9,576), and repaglinide monotherapy ($11,910). These results must be confirmed in larger populations, but they imply that differences in pharmacy costs of repaglinide-metformin therapy are offset by measurable medical cost savings. PMID- 12908216 TI - The effect of drug reimportation on managed care pharmacy. PMID- 12908217 TI - Health politics or health policy? PMID- 12908218 TI - Malpractice reform: one state's example. AB - The medical malpractice liability crisis has resulted in soaring liability costs, the closing of major malpractice insurance companies, and many doctors abandoning the performance of high-risk procedures or medical practice altogether. The article explores how medical malpractice liability was addressed in the State of Pennsylvania, as well as recent attempts at tort reform by the federal government. PMID- 12908219 TI - Interview with Phil Robinson, FACHE, CEO, JFK Medical Center, Palm Beach County, Florida. PMID- 12908220 TI - Diagnosing risk: strategies for preventing unexpected job loss. PMID- 12908221 TI - The art of competing against yourself: are you a cannibal? PMID- 12908222 TI - Challenges and opportunities facing health administration practice and education. PMID- 12908223 TI - Healthcare reform through rationing. AB - Currently, the U.S. government cannot afford to pay for or ensure access to healthcare without limitations for all its citizens, as healthcare has become so expensive. Therefore, implementation of a rationing program is likely in the future for the U.S. healthcare system. Key stakeholders in such an initiative include citizens, healthcare providers, and the government. Americans expect to have unlimited access to care, but they do not like its high cost. Healthcare providers focus on giving the highest quality of care, with costs as a secondary concern. As a payer, the government wants to provide broader access to healthcare and, at the same time, reduce healthcare expenditures. The current debate over healthcare rationing surrounds three key questions: (1) can medically necessary procedures be ethically denied, (2) do Americans have a right to unlimited access to healthcare regardless of the costs, and (3) what is the limit on the availability of healthcare resources for those with the ability to pay. Proposals for rationing include developing a program similar to the Oregon Health Plan, using the quality-adjusted life years method, promoting self-rationing to consumers, or encouraging healthcare providers to actively and openly participate in bedside rationing. These proposals are evaluated based on their economic efficiency, fairness, and utility. The Oregon Health Plan is recommended among these proposals as it benefits the stakeholders through the explicit and equitable allocation of scarce healthcare resources. PMID- 12908224 TI - Robot-assisted surgery: the future is here. AB - According to L. Wiley Nifong, director of robotic surgery at East Carolina University's Brody School of Medicine, "Nationally, only one-fourth of the 15 million surgeries performed each year are done with small incisions or what doctors call 'minimally invasive surgery'." Robots could raise that number substantially (Stark 2002). Currently, healthcare organizations use robot technology for thoracic, abdominal, pelvic, and neurological surgical procedures. Minimally invasive surgery reduces the amount of inpatient hospital days, and the computer in the system filters any hand tremors a physician may have during the surgery. The use of robot-assisted surgery improves quality of care because the patient experiences less pain after the surgery. Robot-assisted surgery demonstrates definite advantages for the patient, physician, and hospital; however, healthcare organizations in the United States have yet to acquire the technology because of implementation costs and the lack of FDA (Food and Drug Administration) approval for using the technology for certain types of heart procedures. This article focuses on robot-assisted surgery advantages to patients, physicians, and hospitals as well as on the disadvantages to physicians. In addition, the article addresses implementation costs, which creates financial hurdles for most healthcare organizations; offers recommendations for administrators to embrace this technology for strategic positioning; and enumerates possible roles for robots in medicine. PMID- 12908225 TI - A model for improving the quality and timeliness of compensation and pension examinations in VA facilities. AB - In response to external and internal customer dissatisfaction and in anticipation of markedly higher volumes of examination requests, the Department of Veterans Affairs (VA) produced an eight-month facilitated quality-improvement project designed to improve the quality and timeliness of compensation examination processing. To determine whether participation in the project was associated with better outcomes and to identify team characteristics that were associated with high performance, we obtained centrally collected facility-level performance measures on quality and timeliness of the examinations. To determine factors associated with team success, we compared measures of leadership support reported by teams with high and low performance outcomes. Thirty teams representing 34 VA medical centers and 22 Veterans Benefits Administration's regional offices participated in the project. Monthly volumes were significantly higher for participating teams, and volumes increased significantly over time for both groups. At the beginning of the project, examination timeliness was substantially worse for participating teams (34.1 versus 29.9 days, p = .03); by the end, participants had better performance (28.5 versus 30.3 days, p = .00). Quality measures were maintained. By the end of the project, high performers reported improved leadership, frontline support, resource availability, alignment with strategic goals, and leadership mandate when compared to performance at the beginning of the project; low performers reported the opposite. These results suggest that the principles of clinical improvement can be applied successfully to teach teams how to achieve process improvements within a large healthcare organization. Visible, ongoing support by leadership and alignment of project objectives with strategic goals are associated with improved project outcomes. PMID- 12908226 TI - Adapting manufacturing-based Six Sigma methodology to the service environment of a radiology film library. PMID- 12908227 TI - Hydrogen storage in carbon nanotubes. AB - The article gives a comprehensive overview of hydrogen storage in carbon nanostructures, including experimental results and theoretical calculations. Soon after the discovery of carbon nanotubes in 1991, different research groups succeeded in filling carbon nanotubes with some elements, and, therefore, the question arose of filling carbon nanotubes with hydrogen by possibly using new effects such as nano-capillarity. Subsequently, very promising experiments claiming high hydrogen storage capacities in different carbon nanostructures initiated enormous research activity. Hydrogen storage capacities have been reported that exceed the benchmark for automotive application of 6.5 wt% set by the U.S. Department of Energy. However, the experimental data obtained with different methods for various carbon nanostructures show an extreme scatter. Classical calculations based on physisorption of hydrogen molecules could not explain the high storage capacities measured at ambient temperature, and, assuming chemisorption of hydrogen atoms, hydrogen release requires temperatures too high for technical applications. Up to now, only a few calculations and experiments indicate the possibility of an intermediate binding energy. Recently, serious doubt has arisen in relation to several key experiments, causing considerable controversy. Furthermore, high hydrogen storage capacities measured for carbon nanofibers did not survive cross-checking in different laboratories. Therefore, in light of today's knowledge, it is becoming less likely that at moderate pressures around room temperature carbon nanostructures can store the amount of hydrogen required for automotive applications. PMID- 12908228 TI - Science and applications of single-nanotube Raman spectroscopy. AB - A review is presented of the resonance Raman spectra from individual isolated single-wall carbon nanotubes (SWNTs). A brief summary is given of how the measurements are made. Why the resonance Raman effect allows single-carbon nanotube spectra to be observed easily and under normal operating conditions is summarized. The important structural information that is provided by single nanotube spectroscopy using one laser line is discussed, and what else can be learned from tunable laser experiments is reviewed. Particular attention is given to the determination of the nanotube diameter and of the energy of its van Hove singularities Eii. Applications of single-nanotube spectroscopy are emphasized, such as measurements of isolated SWNTs connected with circuit-based samples and of isolated SWNTs mounted on an atomic force microscope tip. A critical assessment of the opportunities and limitations of the resonance Raman method for structural (n, m) identification is presented. The trigonal warping effect, which is central to the (n, m) identification in resonance Raman spectroscopy, is discussed in simple terms, and the importance of this effect in nanotube science and applications is reviewed. PMID- 12908229 TI - Carbon nanotube field emitter. AB - Recently, carbon nanotubes (CNTs), possessing excellent properties as field emitters, are attracting considerable attention as electron emitters of a cold cathode. In this review article, field emission phenomena of carbon nanotubes with various morphologies and surfaces (clean surface or adsorbed molecules on it) revealed by field emission microscopy are first described. Then, the main subject of this article, application of CNTs as electron sources in display devices is reviewed. Other electric devices utilizing CNT-field emitters are also presented. PMID- 12908230 TI - Carbon arc plasma as a source of nanotubes: emission spectroscopy and formation mechanism. AB - Diagnostics of carbon arc plasma by optical emission spectroscopy during the synthesis of carbon nanotubes is reviewed. Spatial distributions of temperature and C2 radicals in different plasmas are presented. The influence of gas pressure, anode composition, and reaction environment is discussed. Mechanisms of carbon nanotube formation are reviewed, with an emphasis on surface diffusion processes and catalytic effects. PMID- 12908231 TI - Iron catalyst chemistry in modeling a high-pressure carbon monoxide nanotube reactor. AB - The high-pressure carbon monoxide (HiPco) technique for producing single-wall carbon nanotubes (SWNTs) is analyzed with the use of a chemical reaction model coupled with flow properties calculated along streamlines, calculated by the FLUENT code for pure carbon monoxide. Cold iron pentacarbonyl, diluted in CO at about 30 atmospheres, is injected into a conical mixing zone, where hot CO is also introduced via three jets at 30 degrees with respect to the axis. Hot CO decomposes the Fe(CO)5 to release atomic Fe. Then iron nucleates and forms clusters that catalyze the formation of SWNTs by a disproportionation reaction (Boudouard) of CO on Fe-containing clusters. Alternative nucleation rates are estimated from the theory of hard sphere collision dynamics with an activation energy barrier. The rate coefficient for carbon nanotube growth is estimated from activation energies in the literature. The calculated growth was found be about an order of magnitude greater than measured, regardless of the nucleation rate. A study of cluster formation in an incubation zone prior to injection into the reactor shows that direct dimer formation from Fe atoms is not as important as formation via an exchange reaction of Fe with CO in FeCO. PMID- 12908232 TI - Effects of carbonyl bond, metal cluster dissociation, and evaporation rates on predictions of nanotube production in high-pressure carbon monoxide. AB - The high-pressure carbon monoxide (HiPco) process for producing single-wall carbon nanotubes (SWNTs) uses iron pentacarbonyl as the source of iron for catalyzing the Boudouard reaction. Attempts using nickel tetracarbonyl led to no production of SWNTs. This paper discusses simulations at a constant condition of 1300 K and 30 atm in which the chemical rate equations are solved for different reaction schemes. A lumped cluster model is developed to limit the number of species in the models, yet it includes fairly large clusters. Reaction rate coefficients in these schemes are based on bond energies of iron and nickel species and on estimates of chemical rates for formation of SWNTs. SWNT growth is measured by the conformation of CO2. It is shown that the production of CO2 is significantly greater for FeCO because of its lower bond energy as compared with that of NiCO. It is also shown that the dissociation and evaporation rates of atoms from small metal clusters have a significant effect on CO2 production. A high rate of evaporation leads to a smaller number of metal clusters available to catalyze the Boudouard reaction. This suggests that if CO reacts with metal clusters and removes atoms from them by forming MeCO, this has the effect of enhancing the evaporation rate and reducing SWNT production. The study also investigates some other reactions in the model that have a less dramatic influence. PMID- 12908234 TI - Oxidative properties and chemical stability of fluoronanotubes in matrixes of binary inorganic compounds. AB - The chemical stability of fluoronanotubes in selected solid inorganic matrixes has been studied by initially mixing and mechanically grinding the components and subsequently heating them at temperatures ranging from 35 to 600 degrees C. The inorganic compounds selected for matrixes included halides (KBr, KI, Lil, LiBr, LiCl, NaCl, Znl2), oxides (Li2O, Fe2O3, PbO, MnO), lithium peroxide (Li2O2), potassium superoxide (KO2), sulfides (Li2S and ZnS), zinc selenide (ZnSe), lithium nitride (Li3N), and aluminum phosphide (AIP). Solid products, resulting from the proceeding chemical reactions, were analyzed by X-ray diffraction, Raman spectroscopy, and SEM/EDX elemental analysis. Gaseous and volatile products were identified with the help of the TGA/MS technique. Experimental data presented in this paper provide clear evidence that fluoronanotubes are not chemically inert toward the solid matrixes studied and exhibit significant oxidative properties in the redox reactions occurring under various temperatures, depending on the nature of the inorganic compound. PMID- 12908233 TI - The role of surfactant adsorption during ultrasonication in the dispersion of single-walled carbon nanotubes. AB - The ionic surfactant-assisted dispersion of single-walled carbon nanotubes in aqueous solution has been studied by Raman and fluorescent spectroscopy during ultrasonic processing. During the process, an equilibrium is established between free individuals and aggregates or bundles that limits the concentration of the former that is possible. This equilibrium is a function of free sodium dodecyl sulfate concentration. At surfactant concentrations below this value, fluorescence is shifted to a lower energy due to an increase in micropolarity from water association at the nanotube surface. The mechanism of dispersion is postulated as the formation of gaps or spaces at the bundle ends in the high shear environment of the ultrasonicated solution. Surfactant adsorption and diffusion then propagate this space along the bundle length, thereby separating the individual nanotube. The former is found to be controlling, with the use of a derived kinetic model for the dispersion process and extraction of the characteristic rate of nanotube isolation. PMID- 12908235 TI - Low-temperature synthesis of large-area CNx nanotube arrays. AB - Well-aligned nitrogen-doped multiwall carbon nanotube arrays have been successfully grown over large areas on quartz and silicon wafers by floating catalyst chemical vapor deposition at low temperatures (600 degrees C). These nitrogen-including nanotubes, derived from pyridine-ferrocene mixtures, have smaller outer diameters but larger inner diameters compared with carbon nanotubes grown from a xylene-ferrocene mixture under similar conditions. The N-doped nanotubes exhibit bamboo-like structures in the core. Elemental analysis and electron energy loss spectroscopy analysis show that the as-prepared nanotubes contain as much as 2.62 wt.% N, with most of the N concentrated in the inner few shells of the nanotube. Such large-scale arrays of well-aligned N-doped nanotubes on silicon wafers have a current density as high as 23.8 mA/cm2 at an applied electric field of 17 V/micron, which can be further improved by patterning the tubes and coating the silicon substrate with a conductive thin metal film for the fabrication of flat panel displays. PMID- 12908236 TI - Synthesis and thermoelectric power of nitrogen-doped carbon nanotubes. AB - We have previously shown that high-purity multiwalled carbon nanotubes (pristine MWNTs) can be prepared from a mixture of xylene-ferrocene (99 at% C:1 at% Fe) inside a quartz tube reactor operating at approximately 700 degrees C. In a similar process, approximately 3 g of melamine (C3H6N6) was introduced during the growth of MWNTs to prepare nitrogen-doped nanotubes. The structural and electronic properties of nitrogen-doped MWNTs were determined by scanning electron microscopy, high-resolution transmission electron microscopy (HRTEM), electron energy loss spectroscopy (EELS), and thermopower measurements. The individual nitrogen-doped nanotube exhibits a bamboo-like structure and comprises 6-16 tube walls, as evidenced by HRTEM studies. The EELS measurements yielded an average nitrogen content of approximately 5 at% in the doped tubes. The thermoelectric power data of nitrogen-doped MWNTs remained negative even after exposure to oxygen for an extended period of time, suggesting that nitrogen doping of MWNTs renders them n-type, consistent with scanning tunneling spectroscopic studies on similar nanotubes. PMID- 12908237 TI - Production and characterization of polymer nanocomposites with highly aligned single-walled carbon nanotubes. AB - We report the production and characterization of polymer nanocomposites with single-walled carbon nanotubes having improved mechanical properties and exceptional nanotube alignment. High-pressure carbon monoxide nanotubes (HiPco) were efficiently distributed in polystyrene (PS) and polyethylene (PE) with a twin-screw compounder. Nanotube concentrations were 1, 5, 10, and 20 wt% in PE composites and 0.7 wt% in PS composites. PE composites were melt-spun into fibers to achieve highly aligned nanotubes. Polarized Raman spectroscopy shows that the degree of alignment increases with decreasing fiber diameter and decreases with increasing nanotube loading. The orientation distribution function of a 1 wt% HiPco/PE composite had a full width at half-maximum of approximately 5 degrees. The elastic modulus increases up to 450% relative to PE fibers for 20 wt% nanotube loading at an intermediate fiber diameter of 100 microns. PMID- 12908238 TI - Mesoporous silicates as nanoreactors for carbon nanotube production in the absence of transition metal catalysts. AB - Carbon nanotubes were produced from either a template or the polymer-filled pore systems of mesoporous silicates of various structures and dimensions by heat treatment in the absence of air. Successful synthesis was done when the template molecules contained little or no oxygen. For SBA-15 material, where the structure directing molecule used for synthesis of mesoporous silicate was polyethylene oxide-polypropylene oxide-polyethylene oxide triblock copolymer, no carbon nanostructures were formed. A peculiar carbon nanostructure was generated from the template for pore expanded MCM-41. To demonstrate carbon nanotube formation from polymer in the mesoporous silicates, the mesopores of MCM-41, MCM-48, and SBA-15 silicates were filled with divinyl-benzene polymer and then graphitized at 1300 K. The polymer was successfully transformed into carbon nanotubes for the MCM class silicate but not the SBA-15 silicate. PMID- 12908239 TI - Purification process for vertically aligned carbon nanofibers. AB - Individual, free-standing, vertically aligned multiwall carbon nanotubes or nanofibers are ideal for sensor and electrode applications. Our plasma-enhanced chemical vapor deposition techniques for producing free-standing and vertically aligned carbon nanofibers use catalyst particles at the tip of the fiber. Here we present a simple purification process for the removal of iron catalyst particles at the tip of vertically aligned carbon nanofibers derived by plasma-enhanced chemical vapor deposition. The first step involves thermal oxidation in air, at temperatures of 200-400 degrees C, resulting in the physical swelling of the iron particles from the formation of iron oxide. Subsequently, the complete removal of the iron oxide particles is achieved with diluted acid (12% HCl). The purification process appears to be very efficient at removing all of the iron catalyst particles. Electron microscopy images and Raman spectroscopy data indicate that the purification process does not damage the graphitic structure of the nanotubes. PMID- 12908240 TI - Spectroscopic probing of organic molecules encapsulated in functionalized carbon nanotubes in solution. AB - Pyrene was introduced into cavities in functionalized single-walled and multiple walled carbon nanotubes to be used as a molecular probe in the study of encapsulation. The solubility of these materials in common organic solvents allowed solution-phase absorption and emission spectroscopic measurements. The results, which are consistent with the formation of pyrene excimer, are explained in terms of high local pyrene concentrations and perhaps pyrene microcrystals inside the carbon nanotube cavities. The fluorescence decay results show that there is significant quenching of pyrene excited states by the hosting carbon nanotubes. PMID- 12908241 TI - Raman characterization of single-walled nanotubes of various diameters obtained by catalytic disproportionation of CO. AB - Single-walled carbon nanotubes prepared by disproportionation of CO over Co Mo/SiO2 catalysts have been characterized by Raman spectroscopy, using several excitation energies. By varying the reaction temperature, different ranges of nanotube diameter were obtained. The average diameter of a single-walled nanotube produced at 750 degrees C was 0.9 nm, while it increased up to about 1.5 nm when the synthesis was conducted at 950 degrees C. The analysis of the Raman spectra obtained with a range of laser excitation energies not only gives a definite description of the single-walled nanotubes diameters but also helps differentiate the metallic or semiconducting character of the samples. This analysis can be done by comparing the experimental data with calculated gap energies as a function of nanotube diameter as well as comparing the relative intensity of bands centered at 50-60 cm-1 lower than the tangential G mode. The analysis of this feature, which can be fitted with a Breit-Wigner-Fano line, offers a method for distinguishing between metallic and semiconducting single-walled carbon nanotubes. PMID- 12908243 TI - Temperature dependence of electron-to-lattice energy transfer in single-wall carbon nanotube bundles. AB - The electron-phonon coupling strength in single-wall carbon nanotube (SWNT) bundles has been studied directly in the time domain by femtosecond time-resolved photoelectron spectroscopy. We have measured the dependence of H(Te, Tl), the rate of energy transfer between the electronic system and the lattice as a function of electron and lattice temperatures Te and Tl. The experiments are consistent with a T5 dependence of H on the electron and lattice temperatures, respectively. The results can be related to the e-ph mass enhancement parameter lambda. The experimentally obtained value of for lambda/[symbol: see text] D2, where [symbol: see text] D is the Debye temperature, suggests that e-ph scattering times at the Fermi level of SWNT bundles can be exceptionally long, exceeding 1.5 ps at room temperature. PMID- 12908242 TI - High-pressure Raman study of debundled single-walled carbon nanotubes. AB - We report the pressure dependence for the radial (omega R) and tangential (omega T) band frequencies in debundled single-walled carbon nanotubes (SWNTs) derived from laser-synthesized SWNT bundles. As previously described, a chemical procedure was used to prepare debundled SWNTs from as-prepared, large SWNT bundles. The normalized pressure coefficient for omega R in the debundled sample was compared with the corresponding value in the bundled sample to quantify the strength of van der Waals interactions between tubes in these nanotube materials. Furthermore, the pressure dependences for the radial (omega R) and tangential (omega T) band frequencies in debundled tubes were also compared with corresponding dependences predicted for isolated SWNTs, obtained with generalized tight binding molecular dynamic (GTBMD) simulations described in our previous work. The results presented here collectively suggest that the van der Waals interaction is still strong in the debundled sample studied here, which contained predominantly small bundles of SWNTs rather than isolated tubes. PMID- 12908244 TI - Double-walled carbon nanotubes in composite powders. AB - Double-walled carbon nanotubes (DWNTs) may be interesting in many applications since the outer wall would provide an interface with the rest of the system, without modifying the inner wall. CNT-Fe/Fe3C-Al2O3 composite powders containing carbon nanotubes (CNTs) (65% of which are DWNTs) are prepared by reduction of an oxide solid solution in a H2-CH4 gas mixture. The powders and CNTs are studied by both local and macroscopical techniques. The influence of the reducing atmosphere composition and of the dwell time at 1050 degrees C is studied. There is a 6-fold increase in CNT content upon the increase in the CH4 content from 3 to 30 mol.%, but the formation of undesirable carbon nanofibers can also be promoted. A CH4 content of 12-18 mol.% is adapted for the particular iron content in these powders. Increasing the dwell time at 1050 degrees C results in the formation of CNTs with more walls. PMID- 12908245 TI - Filling single-wall carbon nanotubes with d- and f-metal chloride and metal nanowires. AB - Nanowires of magnetic metals (Fe, Co, Ho, Gd) have been synthesized inside the hollow interiors of single-wall carbon nanotubes (SWNTs) by filling SWNTs with precursor metal chlorides and subsequent reduction. SWNTs have been filled by either the melt-phase sealed-tube reaction or a solution-phase method. Among the metal chlorides investigated in this study, HoCl3 and GdCl3 filled the SWNTs to a significantly higher extent. The nanowires have been imaged by transmission electron microscopy (TEM), high-resolution transmission electron microscopy, and scanning transmission electron microscopy (STEM). X-ray energy dispersive spectroscopy carried out in conjunction with TEM and STEM confirmed the presence of metal chloride and metal nanowires. PMID- 12908246 TI - Ni and Ni/Pt filling inside multiwalled carbon nanotubes. AB - Multiwalled carbon nanotubes are grown by microwave plasma chemical vapor deposition with CH4 and H2 as precursor gases. Ni and Ni/Pt electroplated layers are used as catalysts for the synthesis of the tubes. We observe that a very efficient filling of the tubes takes place with Ni. In some cases Ni/Pt filling is also observed inside the tubes. High-resolution transmission electron microscopy (HRTEM) studies, coupled with energy-dispersive X-ray analyses of the tubes, indicate Ni nanorods with a highly symmetrical cylindrical structure. The diameter of the cylindrical nanorods is on the order of 40 nm, and their length is 660 nm. The nano area diffraction pattern of the nanorods reveals the cubic structure of nickel, and electron diffraction spots corresponding to (111), (200), (220) planes are evident. The lattice constant of Ni measured from the diffraction spots was found to be 0.347 +/- 0.0013 nm. This should be compared with 0.352 nm, the value of "a" in bulk Ni. The decrease in the lattice constant may be due to the strain experienced inside the tubes. Raman spectroscopy shows the typical signature of the tangential breathing mode present in the tubes at 1580 cm-1 that shifts to a new position when the C12 is replaced by 13C. The shift, however, is too small and is difficult to explain on the basis of mass difference. HRTEM experiments indicate the presence of Ni3C in the samples dominantly in the interfacial region. PMID- 12908247 TI - Ballistic conduction in multiwalled carbon nanotubes. AB - The electrical transport in multiwalled carbon nanotubes is shown to be ballistic at room temperature with mean free paths on the order of tens of microns. The measurements are performed both in air and in the transmission electron microscope by contacting the free end of a nanotube pointing out of a fiber to a liquid metal and measuring the dependence of the nanotube resistance between the contacts. For a specific representative nanotube the resistance per unit length is found to be Rt = 31 +/- 61 omega/micron and the contact resistance with the liquid metal, Rc = 165 +/- 55 omega microns, corresponding to a mean free path l = 200 microns. Current-to-voltage characteristics are in accord with the electronic structure. The nanotubes survive high currents (up to 1 mA, i.e., current density on the order of 10(9) A/cm2). In situ electron microscopy shows that a relatively large fraction of the nanotubes do not conduct (even at high bias), consistent with the existence of semiconducting nanotubes. Discrepancies with other measurements are most likely due to damage caused to the outer layer(s) of the nanotubes during processing. The measured mean free path of clean, undamaged arc-produced multiwalled carbon nanotubes is several orders of magnitude greater than that for metals, making this perhaps the most significant property of carbon nanotubes. PMID- 12908248 TI - Field emission properties of carbon nanotubes coated with boron nitride. AB - Field emission properties of carbon nanotubes coated with a single layer of boron nitride are calculated using the first-principles pseudopotential method. At lower bias voltage, the emission current of the coated nanotube is comparable to that of the bare carbon nanotube and is dominated by the contribution from localized states at the tip of the tube. At higher voltage, newly generated hybridized states between the carbon nanotube tip and the even-membered boron nitride rings contribute significantly to the emission current because they experience a low tunneling barrier compared with the bare carbon nanotube case. Our results suggest that the insulator coating can, besides protecting the nanotube tip from the attack of gas molecules, substantially enhance the field emission current. PMID- 12908249 TI - Effect of interlayer potential on mechanical deformation of multiwalled carbon nanotubes. AB - A study on the modeling and simulation of interlayer interaction in the multiwalled carbon nanotube (MWCNT) system is presented. We use an interlayer Morse potential previously developed from a local density approximation (LDA) treatment of a bilayer of graphite. We have fit this Morse potential to experimental high-pressure compressibility data for graphite and to a more extensive LDA equation of state (EOS) for graphite, and excellent agreement is observed. We employ this potential to treat the interlayer mechanics of MWCNTs, where the MWCNT is so highly deformed that interlayer separation well below approximately 0.34 nm, such as down to approximately 0.26 nm, is occurring. This, to our knowledge, is the first treatment that attempts to account for deformations that have the layers approaching each other at very high local (interlayer) stress levels. Since evaluating the interlayer potential for a large MWCNT system is computationally intensive, a continuum simulation approach is proposed that saves on computational time and thus on cost. Comparisons with experimental results of buckled and highly kinked MWCNTs are presented. PMID- 12908250 TI - Quantum-chemical model evaluations of thermodynamics and kinetics of oxygen atom additions to narrow nanotubes. AB - This paper reports a computational study of oxygen additions to narrow nanotubes, a problem frequently studied with fullerenes. In fact, fullerene oxides were the first observed fullerene derivatives, and they have naturally attracted the attention of both experiment and theory. C60O had represented a long-standing case of experiment-theory disagreement, and there has been a similar problem with C60O2. The disagreement has been explained by kinetic rather than thermodynamic control. In this paper a similar computational approach is applied to narrow nanotubes. Recently, very narrow nanotubes have been observed with a diameter of 5 A and even with a diameter of 4 A. It has been supposed that the narrow nanotubes are closed by fragments of small fullerenes like C36 or C20. In this report we perform calculations for oxygen additions to such model nanotubes capped by fragments of D2d C36, D4d C32, and Ih C20 fullerenic cages (though the computational models have to be rather short). The three models have the following carbon contents: C84, C80, and C80. Both thermodynamic enthalpy changes and kinetic activation barriers for oxygen addition to six selected bonds are computed and analyzed. The lowest isomer (thermodynamically the most stable) is never of the 6/6 type, that is, the enthalpically favored structures are produced by oxygen additions to the nanotube tips. Interestingly enough, the lowest energy isomer has, for the D2d C36 and D4d C32 cases, the lowest kinetic activation barrier as well. PMID- 12908251 TI - Elastic buckling of multiwall carbon nanotubes under high pressure. AB - This paper studies elastic buckling of individual multiwall carbon nanotubes under radial pressure. The analysis is based on a multiple-elastic-shell model in which each of the concentric tubes of a multiwall carbon nanotube is described as an individual elastic shell. According to their radius-to-thickness ratios, the multiwall carbon nanotubes discussed here are classified into three types: thin, thick, and (almost) solid. The critical pressure for elastic buckling is calculated for examples of all three types. It is found that a thin N-wall nanotube (defined by a radius-to-thickness ratio larger than 4) is approximately equivalent to a single-layer elastic shell whose effective bending stiffness and thickness are N times the effective bending stiffness and thickness of single wall carbon nanotubes. Based on this result, an approximate method is suggested for replacing the problematic multiwall nanotube of many layers with a multilayer elastic shell of fewer layers. In particular, the critical pressure predicted by the present model is in good agreement with known experimental results. PMID- 12908253 TI - Nanometer-scale organic thin film transistors from self-assembled monolayers. AB - A survey of the most interesting results on nanometer-scale organic thin film transistors (nano-OTFT) is presented. Additionally, we discuss our recent results on the properties of end-group functionalized organic self-assembled monolayers and on their use in the fabrication of nanometer-scale field-effect transistors. Nanometer-scale organic transistors (channel length 30 nm) were fabricated, with a self-assembled monolayer as gate insulator. The carrier transport in these transistors, as a function of the channel length, was investigated, and a transition from a dispersive to a ballistic transport at a channel length of 200 nm was observed. On a molecular scale, alkyl monolayers functionalized at their omega-ends by aromatic moieties were prepared. A high anisotropic conductivity in molecular insulator/semiconductor heterostructures of monolayer thickness was observed. These molecular architectures provide a basis for the building blocks of molecular transistors. PMID- 12908254 TI - Magnetic nanostructures and materials in magnetic random access memory. AB - The advances in magnetic random access memory provide a remarkable showcase for the rapid development and application of nanodevices. Several aspects of state-of the-art magnetic nanoscience and nanotechnology are developed and utilized in this single device. Current magnetic random access memory design is built upon the discovery and understanding of physics issues such as giant magnetoresistance, spin-dependent tunneling, exchange bias, and magnetic anisotropy in small elements. Successful magnetic random access memory development requires future research in some of the key areas involving nanotechnology. For example, the uniformity of the barrier thickness across the entire device and magnetic switching stability of a nanosized element are challenging issues that lie ahead. The spin degree of freedom of electrons is an added dimension that is both unique and useful in electronic transport and information technology. Nonvolatile, high-density, high-speed, and low-power magnetic random access memory is one of the first examples of the application of spintronics. PMID- 12908252 TI - Issues of nanoelectronics: a possible roadmap. AB - In this review, we will discuss a possible roadmap in scaling a nanoelectronic device from today's CMOS technology to the ultimate limit when the device fails. In other words, at the limit, CMOS will have a severe short channel effect, significant power dissipation in its quiescent (standby) state, and problems related to other essential characteristics. Efforts to use structures such as the double gate, vertical surround gate, and SOI to improve the gate control have continually been made. Other types of structures using SiGe source/drain, asymmetric Schottky source/drain, and the like will be investigated as viable structures to achieve ultimate CMOS. In reaching its scaling limit, tunneling will be an issue for CMOS. The tunneling current through the gate oxide and between the source and drain will limit the device operation. When tunneling becomes significant, circuits may incorporate tunneling devices with CMOS to further increase the functionality per device count. We will discuss both the top down and bottom-up approaches in attaining the nanometer scale and eventually the atomic scale. Self-assembly is used as a bottom-up approach. The state of the art is reviewed, and the challenges of the multiple-step processing in using the self assembly approach are outlined. Another facet of the scaling trend is to decrease the number of electrons in devices, ultimately leading to single electrons. If the size of a single-electron device is scaled in such a way that the Coulomb self-energy is higher than the thermal energy (at room temperature), a single electron device will be able to operate at room temperature. In principle, the speed of the device will be fast as long as the capacitance of the load is also scaled accordingly. The single-electron device will have a small drive current, and thus the load capacitance, including those of interconnects and fanouts, must be small to achieve a reasonable speed. However, because the increase in the density (and/or functionality) of integrated circuits is the principal driver, the wiring or interconnects will increase and become the bottleneck for the design of future high-density and high-functionality circuits, particularly for single-electron devices. Furthermore, the massive interconnects needed in the architecture used today will result in an increase in load capacitance. Thus for single-electron device circuits, it is critical to have minimal interconnect loads. And new types of architectures with minimal numbers of global interconnects will be needed. Cellular automata, which need only nearest-neighbor interconnects, are discussed as a plausible example. Other architectures such as neural networks are also possible. Examples of signal processing using cellular automata are discussed. Quantum computing and information processing are based on quantum mechanical descriptions of individual particles correlated among each other. A quantum bit or qubit is described as a linear superposition of the wave functions of a two-state system, for example, the spin of a particle. With the interaction of two qubits, they are connected in a "wireless fashion" using wave functions via quantum mechanical interaction, referred to as entanglement. The interconnection by the nonlocality of wave functions affords a massive parallel nature for computing or so-called quantum parallelism. We will describe the potential and solid-state implementations of quantum computing and information, using electron spin and/or nuclear spin in Si and Ge. Group IV elements have a long coherent time and other advantages. The example of using SiGe for g factor engineering will be described. PMID- 12908255 TI - Nanostructured ceramics for biomedical implants. AB - Recent progress in the synthesis, characterization, and biological compatibility of nanostructured ceramics for biomedical implants is reviewed. A major goal is to develop ceramic coating technology that can reduce the friction and wear in mating total joint replacement components, thus contributing to their significantly improved function and longer life span. Particular attention is focused on the enhancement of mechanical properties such as hardness, toughness, and friction coefficient and on the bioactivity as they pertain to the nanostructure of the material. The development of three nanostructured implant coatings is discussed: diamond, hydroxyapatite, and functionally graded metalloceramics based on the Cr-Ti-N ternary system. Nanostructured diamond produced by chemical vapor deposition (CVD) techniques and composed of nano-size diamond grains have particular promise because of the combination of ultrahigh hardness, improved toughness over conventional microcrystalline diamond, low friction, and good adhesion to titanium alloys. Nanostructured processing applied to hydroxyapatite coatings is used to achieve the desired mechanical characteristics and enhanced surface reactivity and has been found to increase osteoblast adhesion, proliferation, and mineralization. Finally, nanostructured metalloceramic coatings provide continuous variation from a nanocrystalline metallic bond at the interface to the hard ceramic bond on the surface and have the ability to overcome adhesion problems associated with ceramic hard coatings on metallic substrates. PMID- 12908256 TI - Lithographic fine-tuning of vertical cavity surface emitting laser-pumped two dimensional photonic crystal lasers. AB - Lithographic tuning of operating wavelengths in a photonic crystal laser array is demonstrated. The photonic crystal lattice constant is varied by 2 nm between elements of the array, and a wavelength spacing of approximately 4 nm is achieved. PMID- 12908257 TI - Photostable luminescent nanoparticles as biological label for cell recognition of system lupus erythematosus patients. AB - In this article, we report a method for cell recognition of system lupus erythematosus (SLE) patients that uses photostable luminescent nanoparticles as biological labels. The luminescent silica nanoparticles are prepared with a water in-oil microemulsion (W/O) technique. The silica network is produced by the controlled hydrolysis of tetraethylorthosilicate (TEOS) in water nanodroplets with the initiation of ammonia (NH3.H2O). A luminescent compound, tris(2,2' bipyridyl)dichlororuthenium(II)hexahydrate [Ru(II)(bpy)3]2+, is doped inside as a luminescent signaling element, and the most appropriate dye concentration for the preparation of the nanoparticles with a size of 28 +/- 4 nm has been determined. The luminescent silica nanoparticles are covalently immobilized with goat anti human immunoglobulin G (IgG), which can recognize SmIgG+ B lymphocytes. We have used antibody-labeled nanoparticles to recognize target SmIgG+ B lymphocytes isolated from the circulating blood of SLE patients. It has been observed that a bioassay based on fluorescent nanoparticles can identify target cells selectively and efficiently. And fluorescent nanoparticle labels also exhibit high photostability. The experiment results have shown that this cell recognition method was an effective one as further proof of the diagnosis of SLE. PMID- 12908258 TI - Fine-pitched microgratings encoded by interference of UV femtosecond laser pulses. AB - Fine-pitched microgratings are encoded on fused silica surfaces by a two-beam laser interference technique employing UV femtosecond pulses from the third harmonics of a Ti:sapphire laser. A pump and prove method utilizing a laser induced optical Kerr effect or transient optical absorption change has been developed to achieve the time coincidence of the two pulses. Use of the UV pulses makes it possible to narrow the grating pitches to an opening as small as 290 nm, and the groove width of the gratings is of nanoscale size. The present technique provides a novel opportunity for the fabrication of periodic nanoscale structures in various materials. PMID- 12908259 TI - Performance of AlGaN/GaN high electron mobility transistors at nanoscale gate lengths. AB - The DC and RF performance of AlGaN/GaN high electron mobility transistors with nanoscale gate lengths is presented. The layer structures were grown by either metal organic chemical vapor deposition or rf plasma-assisted molecular beam epitaxy. Excellent scaling properties were observed as a function of both gate length and width and confirm that these devices are well suited to both high speed switching and power microwave applications. PMID- 12908260 TI - A majority-logic nanodevice using a balanced pair of single-electron boxes. AB - This paper describes a majority-logic gate device that will be useful in developing single-electron integrated circuits. The gate device consists of two identical single-electron boxes combined to form a balanced pair. It accepts three inputs and produces a majority-logic output by using imbalances caused by the input signals; it produces a 1 output if two or three inputs are 1, and a 0 output if two or three inputs are 0. We combine these gate devices into two subsystems, a shift register and an adder, and demonstrate their operation by computer simulation. We also propose a method of fabricating the unit element of the gate device, a minute dot with four coupling arms. We demonstrate by experiments that it is possible to arrange these unit elements on a GaAs substrate, in a self-organizing manner, by means of a process technology that is based on selective-area metalorganic vapor-phase epitaxy. PMID- 12908261 TI - A multi-nano-dot circuit and structure using thermal-noise-assisted tunneling for stochastic associative processing. AB - The single-electron circuit and nanostructure described in this paper are designed for stochastic associative processing, which is an expanded version of ordinary associative memory processing. In stochastic associative processing, the association probability of each stored pattern depends on the similarity between the stored pattern and the input pattern. Such unique processing is useful for sequential stochastic association and for clustering for vector quantization. Conventional single-electron circuits operate only at very low temperature for practical junction capacitance (i.e., 30 K for 0.1 aF) because the charging energy in these circuits is directly related to the tunnel junction capacitance. Our multi-nano-dot circuit and structure operate at room temperature with a junction capacitance around 0.1 aF through tunneling processes assisted by thermal noise. We analyze the operation of this circuit in detail and propose for it a stochastic associative processing operation, where the detection timing of the electron position controls the association probability distribution. PMID- 12908262 TI - A two-stage shift register for clocked Quantum-Dot Cellular Automata. AB - Quantum-Dot Cellular Automata (QCA) is a computational scheme utilizing the position of interacting single electrons within arrays of quantum dots ("cells") to encode and process binary information. Clocked QCA architectures can provide power gain, logic level restoration, and memory features. Using arrays of micron sized metal dots, we experimentally demonstrate operation of a QCA latch-inverter and a two-stage shift register. PMID- 12908263 TI - Spatial patterning of colloidal nanoparticle-based thin film by a combinative technique of layer-by-layer self-assembly and lithography. AB - A novel approach to generating clear patterns of different types of nanoparticles is presented in this paper. Nanoassembly in the vertical direction was combined with planar micropatterning. This provides industrial applications of a popular layer-by-layer method to produce multilayers of polymers, nanoparticles, and proteins organized on the nanometer scale. A thin film of organic polystyrene spheres was first coated on the pretreated silicon wafer with layer-by-layer self assembly. Then a layer of aluminium was deposited on the thin film. A layer of positive photoresist was spun on the surface of aluminum and then illuminated with UV light. The exposed parts of the resist were removed and windows were opened above the aluminum. The subsequent etching removed exposed aluminium and left a polystyrene thin film in the open windows. Oxygen plasma was employed to remove the polystyrene thin film on the bottom. Eventually, aluminum and photoresist were removed and only the desired pattern remained. This approach was also employed for the patterning of the silica nanoparticle thin film, a widely used material in various applications. In this case, wet etching was demonstrated to etch silica particles. A scanning electron microscope was used to produce the image of the pattern. PMID- 12908264 TI - Fundamental device design considerations in the development of disruptive nanoelectronics. AB - In the last quarter of a century silicon-based integrated circuits (ICs) have played a major role in the growth of the economy throughout the world. A number of new technologies, such as quantum computing, molecular computing, DNA molecules for computing, etc., are currently being explored to create a product to replace semiconductor transistor technology. We have examined all of the currently explored options and found that none of these options are suitable as silicon IC's replacements. In this paper we provide fundamental device criteria that must be satisfied for the successful operation of a manufacturable, not yet invented, device. The two fundamental limits are the removal of heat and reliability. The switching speed of any practical man-made computing device will be in the range of 10(-15) to 10(-3) s. Heisenberg's uncertainty principle and the computer architecture set the heat generation limit. The thermal conductivity of the materials used in the fabrication of a nanodimensional device sets the heat removal limit. In current electronic products, redundancy plays a significant part in improving the reliability of parts with macroscopic defects. In the future, microscopic and even nanoscopic defects will play a critical role in the reliability of disruptive nanoelectronics. The lattice vibrations will set the intrinsic reliability of future computing systems. The two critical limits discussed in this paper provide criteria for the selection of materials used in the fabrication of future devices. Our work shows that diamond contains the clue to providing computing devices that will surpass the performance of silicon-based nanoelectronics. PMID- 12908265 TI - Nanocantilever signal transduction by electron transfer. AB - Microfabricated cantilever beams promise to bring about a revolution in the field of chemical, physical, and biological sensor development. The resonance frequency of a microfabricated cantilever shifts sensitively because of mass loading from molecular adsorption. The minimum detectable adsorbed mass on a cantilever sensor can be increased by orders of magnitude by changing the dimensions of the device; smaller and thicker cantilevers offer higher resonance frequency and therefore better mass detection sensitivity. Here we describe micromachined silicon cantilevers that are 0.5 to 4 microns in length, fabricated with the use of a focused ion beam (FIB). In addition, we demonstrate a technique for detection of the cantilever resonance frequency that is based on electron transfer. PMID- 12908266 TI - Phage display antibody-based proteomic device using resonance-enhanced detection. AB - The combination of phage display antibody arrays with a novel nanotransducer technique based on resonant nanoparticles in a nanosandwiched film enables the sensitive parallel screening of proteins. Using the resonance of nanoparticles with their induced mirror dipoles in a thin-film structure, limitations of fluorophores, such as unspecific background and nonvisibility to the eye, can be overcome, thereby leading to an optical signal significantly more sensitive than that of standard colloid techniques. The signal can be both directly observed as a color change of a microdot at the sensor surface and tuned throughout the visible range of the spectrum. Here we report the application of an optical chip using scFv-antibody-antigen interactions. Artificial scFv-antibodies against a variety of proteins, including yeast enzymes and bovine serum albumin (as a standard), were constructed via Phage Display. These scFv-antibodies were then coated onto metal nanoclusters and bound to their antigens that were arrayed as nanodroplets at the resonance layer of the chip. ScFv-Antibody-antigen interaction resulted in a visible array of microdots. Using resonance-enhanced absorption, the absorption signal of the spots was amplified by one to two orders of magnitude (compared to colloid-based techniques). For quantitative analysis, either an 8-micron scanner or a CCD camera (resolution 4 microns) was employed to gain direct-reflection spectra rather than unspecific scatter data (prone to dust and unspecific interaction). Our results demonstrate that this device enables high-throughput proteomics to overcome some limitations of fluorescence, enzyme labels, and colloid techniques. PMID- 12908267 TI - Operation kinetics of a DNA-based molecular switch. AB - The influence of temperature variation, salt concentration, and pH on the operation kinetics of a simple DNA-based molecular switch is investigated. The device shows robust behavior over a wide range of temperatures, pH, and salt concentrations. In particular, the device operates well under physiological conditions. The experimental data can be qualitatively understood in terms of the influence temperature, salt concentration, and pH have on DNA strand interactions. PMID- 12908268 TI - DNA binding to fluorescent ruthenium species released from calcium phosphate/nanoporous silicon structures. AB - This work centers on an analysis of calf thymus DNA binding to emissive Ru complexes which diffuse from biocompatible calcium phosphate/nanoporous silicon films. These nanostructures were characterized by scanning electron microscopy, atomic force microscopy, energy dispersive X-ray analysis, and infrared vibrational spectroscopy. In terms of polynucleotide binding, three different systems were analyzed: (1) an aqueous solution of Ru(phen)(3)2+ (a control); (2) surface-adsorbed Ru(phen)(3)2+ onto undoped calcium phosphate/porous Si/Si in aqueous solution; (3) as-prepared and annealed Ru(phen)3(2+)-doped calcium phosphate/porous Si structures in water. For films with fluorescent Ru originally embedded throughout the film, biphasic diffusion character is found; such behavior is attributed to DNA binding to both surface-bound Ru(phen)(3)2+ and species which originate from deeper regions of the film. PMID- 12908270 TI - Development of organic dye-doped silica nanoparticles for bioanalysis and biosensors. AB - The combination of two silica precursors, tetraethylorthosilicate and phenyltriethoxysilane, were utilized to synthesize organic dye-doped silica nanoparticles. The hydrophobic nature of phenyltriethoxysilane keeps the organic dye in the silica matrix, whereas the hydrophilic tetraethylorthosilicate-formed silica allows the resulting nanoparticles to be dispersed in aqueous solutions. Characterization of the nanoparticles showed that they could be synthesized in the nanometer range with high photostability and minimal dye leakage. The silica matrix of the nanoparticles allows different routes of surface biomolecular modification for biosensor and bioanalysis applications. We have shown different applications of the nanoparticles in bioanalysis and in biosensing. Biotin interaction of avidin-coated nanoparticles can be used for the determination of biotinylated bovine serum albumin, and the immobilization of glutamate dehydrogenase on the nanoparticle surfaces enables the nanoparticles to be used as biosensors for glutamate determination. PMID- 12908269 TI - Covalent attachment of gold nanoparticles to DNA templates. AB - Functionalized gold nanoparticles have been covalently bound to internal, modified sites on double-stranded DNA. Gold nanoparticles coated with mercaptosuccinic acid or thioctic acid were bound to amino-modified thymine bases on double-stranded DNA. Visible absorption spectra, gel electrophoresis, and atomic force microscopy were used to analyze the products. Thiol groups were added to one end of the gold/nanoparticle product, which was then attached to a gold surface. This method has the potential to allow controlled placement of particles with subnanometer precision and to allow attachment of the product to fixed contacts for nanodevice fabrication. PMID- 12908271 TI - Polyelectrolyte microshells as carriers for fluorescent sensors: loading and sensing properties of a ruthenium-based oxygen indicator. AB - A strategy for the design and fabrication of microcapsule-based fluorescent biosensors containing indicators and internal references is described. The rationale for this work is the physical immobilization and chemical separation of assay chemistry for use in biological environments. Using the general approach of depositing oppositely charged species on colloidal micro/nanotemplates, a sensor system employing polyelectrolyte microshells for uptake of functional molecules is proposed, and experiments to demonstrate the feasibility of nanoengineering the sensor properties are described in the context of an oxygen sensor. Methods for immobilization and entrapment of fluorescent indicator and reference dyes are shown, along with the pH dependence of this process. Embedded dyes are shown to be stable and retain their function, as demonstrated with oxygen-sensitivity experiments of loaded microcapsules. Although oxygen sensitivity is presented as an example of a specific application, the overall strategy is likely more generally useful. The work suggests that polyelectrolyte microshells may be used as a platform to develop novel sensors by entrapment of functional materials. PMID- 12908272 TI - Optical spectra of nanowires of Cu and Zn chalcogenides. AB - Nanowires of CuS, ZnS, CuSe, and ZnSe, with diameters in the 5-150-nm range and high aspect ratios, have been prepared with the use of surfactants and characterized by X-ray diffraction and electron microscopy. Electronic absorption spectra and photoluminescence spectra of chalcogenide nanowires have been studied. The spectra show blue-shifts compared with those of bulk samples. PMID- 12908274 TI - Effects of nanometer-thick passivation layers on the mechanical response of thin gold films. AB - The mechanical behavior of freestanding gold membranes 0.5 micron thick with and without passivation layers was studied with a membrane deflection experiment. Membrane width was varied from 2.5 to 20 microns to investigate size effects. The presence of the passivation layer had the effect of reducing the membrane strength. Yield stress, as well as fracture strain and stress, were all found to be significantly lower for the passivated specimens. The residual stress state was found to be significantly larger with passivation, to the degree of generating prestressed cracks at micromachined notches. Membrane width had the greatest effect on the residual stress state with smaller widths having larger residual stress. PMID- 12908273 TI - InAs quantum dots capped by GaAs, In0.4Ga0.6As dots, and In0.2Ga0.8As well. AB - We have fabricated and characterized three types of InAs quantum dots (QDs) with different InxGa1-xAs capping layers. Post-growth atomic force microscopy measurements show that the In0.2Ga0.8As/InAs structure has a smooth surface (dot in-well structure), whereas the In0.4Ga0.6As/InAs structure revealed large QDs with a density similar to that underneath InAs QDs on GaAs (dot-in-dot). With increasing In mole fraction of the capping layer and increasing In0.4Ga0.6As thickness, the energy position of the room-temperature photoluminescence (PL) peak is red-shifted. The quantum dot-in-dot structure emits stronger room temperature PL than does the quantum dot-in-well structure. With a spatially distributed strain in the InAs quantum dot, we have solved the three-dimensional Schrodinger equation by the Green's function theory for the eigenvalues and eigen wave functions. It is concluded that the ground state increases its wave function penetration into the low-barrier InxGa1-xAs capping layer so that its energy position is red-shifted. The reduced PL peak intensity of the dot-in-well (compared with GaAs covered dots) is due to the reduced overlapping between the ground state and the extended states above the GaAs barrier. The overlapping reduction in the dot-in-dot is over compensated for by the reduced relaxation energy (full width at half-maximum), indicating the importance of the sample quality in determining the PL intensity. PMID- 12908275 TI - Engineering in the nanocosmos: Nanorobotics moves kilograms of mass. AB - In recent years Klocke Nanotechnik and partners have developed a new Nanorobotics system that builds a bridge between nanotechnology and classical mechanical engineering. These products combine the advantages of the two technologies: the backlash-free movement of up to 2 kg of load over a stroke of up to 70 mm with a resolution of a few nanometers. This series of positioning modules offers numerous degrees of freedom in the smallest volume for microassembly, interconnection technology, analysis, and reliability testing. Integrated position sensors allow automatic usage, also in rough production technology applications. The repeatability of classical assembly lines is limited to a few microns. Nanorobotics modules allow absolute positioning within 50 nm. PMID- 12908276 TI - Inherently conducting polymer nanostructures. AB - Inherently conducting polymers (ICPs) have been an area of intense interest over the past 30 years, culminating with the award of the 2000 Nobel Prize in Chemistry to MacDiarmid, Heeger and Shirakawa. More recently the unique properties of these materials (e.g., higher conductivity, more rapid discrete electrochemical switching processes) apparent at the nanodimension have become accessible. Significant breakthroughs in synthesis and fabrication of inherently conducting polymers with nanodimensional control have made this possible. This review aims to discuss some of the synthetic approaches researchers have made in an attempt to probe the nano domain as well as some of the property enhancements afforded to these structures. PMID- 12908277 TI - A simple method for the synthesis of silicon carbide nanorods. AB - SiC nanorods were synthesized by a reaction at a temperature of 1200 degrees C, under an argon gas atmosphere, from silicon and amorphous carbon powders mixed by ball milling. The reaction product, which contain SiC nanorods and nanoparticles, has been characterized by high-resolution transmission electron microscopy, X-ray diffraction, and micro-Raman spectroscopy. The synthesized nanorods are more than 1 micron long with a mean diameter of about 10-30 nm. The nanorods possess a well defined crystalline structure with a thin layer of amorphous SiO2 on the surface. Raman shifts of SiC nanorods and the role of structural defects are discussed. PMID- 12908278 TI - Functionalization of carbon nanotubes with bovine serum albumin in homogeneous aqueous solution. AB - Single-walled (SWNTs) and multiple-walled (MWNTs) carbon nanotubes were solubilized via the esterification of nanotube-bound carboxylic acids by oligomeric polyethylene glycol compounds. The water-soluble samples were used as starting materials in reactions with bovine serum albumin (BSA) protein in ambient aqueous solutions. The reaction conditions were designed for thermodynamically favorable transformation from ester to amide linkages, yielding SWNT-BSA and MWNT-BSA conjugates. The results show that the use of soluble starting nanotube materials in an indirect functionalization method represents a valuable approach to the biomodification of carbon nanotubes. PMID- 12908279 TI - Nuclear magnetic resonance of molecular hydrogen trapped in single-walled carbon nanotube bundles. AB - Molecular dynamics of hydrogen trapped in single-walled carbon nanotube bundles was analyzed by nuclear magnetic resonance. The chemical shift of hydrogen was about 5.1 ppm at 293 K, which is similar to that of water. The relaxation time, T1, was about 0.1-0.2 s. Values in this work are comparable to those for hydrogen loaded in silica and a-Si. PMID- 12908280 TI - Possible existence of a higher coverage quasi-one-dimensional phase of argon adsorbed on bundles of single-walled carbon nanotubes. AB - We present results of Ar adsorption isotherms at very low coverages in the first layer and, beyond monolayer completion, on bundles of close-ended single-walled carbon nanotubes. The low coverage results were used to determine the isosteric heat of adsorption and the binding energy of Ar in the groove sites in the first layer. The higher coverage results show evidence of the possible formation of a second-layer groove phase, beyond monolayer completion. Our results for higher coverages are compared with recent computer simulations for this system. PMID- 12908281 TI - Observation of assembly of fluorescent Si nanoparticles under the influence of electric current. AB - A silicon substrate patterned by an oxide is immersed in an alcohol solution of low-doped 1-nm Si nanoparticles. Reverse biasing draws particles to the substrate, mostly along the conducting current paths. Scanning electron and fluorescence microscopy show a tree-like network on the substrate. Avoidance of closed loops and preference for an angle of branching of 90 degrees-120 degrees are observed. The building block of the tree network is not individual particles but spherical particle aggregates approximately 150 nm in diameter. PMID- 12908282 TI - Plasma-enhanced chemical vapor deposition of multiwalled carbon nanofibers. AB - Plasma-enhanced chemical vapor deposition is used to grow vertically aligned multiwalled carbon nanofibers (MWNFs). The graphite basal planes in these nanofibers are not parallel as in nanotubes; instead they exhibit a small angle resembling a stacked cone arrangement. A parametric study with varying process parameters such as growth temperature, feedstock composition, and substrate power has been conducted, and these parameters are found to influence the growth rate, diameter, and morphology. The well-aligned MWNFs are suitable for fabricating electrode systems in sensor and device development. PMID- 12908283 TI - High yield of pure multiwalled carbon nanotubes from the catalytic decomposition of acetylene on in-situ formed cobalt nanoparticles. AB - For the first time, multiwalled carbon nanotubes (MWNTs) could be formed selectively in a high yield, free of any disordered carbon by-product, from the catalytic decomposition of acetylene at 600 degrees C on a CoxMg(1-x)O solid solution. Starting from 1 g of catalytic substrate, 4 g of pure MWNTs were obtained after its dissolution in boiling concentrated HCl, without any additional purification in strongly oxidizing medium, as is required for other methods of nanotube production. In situ reduction of CoO by dihydrogen liberated from acetylene decomposition allows highly divided metal particles to be continuously produced as synthesis proceeds. This is undoubtedly the reason for the good performance of the catalyst and for the ability to produce nanotubes in a narrow diameter range, namely from 10 to 15 nm. With the use of acetylene instead of methane, the synthesis proceeds at low temperature, which prevents the growth of carbon shells, in which the metal particles are generally embedded, decreasing their activity. Because of the very low specific surface area of the catalyst support, the amount of disordered carbon by-product formed is negligible. PMID- 12908284 TI - Mechanical and magnetic properties of nickel-dispersed tetragonal zirconia nanocomposites. AB - Effects of Ni dispersions on microstructure and mechanical properties have been studied for Y2O3-stabilized tetragonal zirconia (Y-TZP)/Ni nanocomposites with Ni dispersion up to 10 vol%. Composites were successfully fabricated by reducing and hot-pressing Y-TZP/NiO powder mixtures. Fracture strength was significantly improved from 1.5 GPa for monolithic Y-TZP to 1.9 GPa for nanocomposites with a small addition of Ni (1-2 vol%). Magnetic properties of Y-TZP/Ni nanocomposites were also investigated. Magnetization curves of Y-TZP/Ni nanocomposites showed typical hysteresis loops of soft magnetic materials, whereas coercivity was much larger than that of pure Ni metal. A new function arising from magnetomechanical effects of metallic Ni is also discussed for the present nanocomposites. PMID- 12908285 TI - Application of Ni:SiO2 nanocomposite to control the carbon deposition on the carbon dioxide reforming of methane. AB - Stable Ni nanoparticles embedded in a mesoporous silica material were used as catalysts for the conversion of methane into synthesis gas. This catalyst has the singular properties of controlling the carbon deposition and deactivation of active sites. A comparative study of our nanocomposites with conventional catalysts showed that impregnation material presented a preferential encapsulation and growth of carbon nanotubes on the metal surface. The impregnated catalyst showed a higher tendency for carbon nanotube and whiskers formation. PMID- 12908286 TI - A Raman study of CdSe and ZnSe nanostructures. AB - Raman studies have been carried out on CdSe nanotubes and ZnSe nanorods produced by surfactant-assisted synthesis. The Raman spectrum of CdSe nanotubes shows modes at 207.5 and 198 cm-1; the former arises from the longitudinal optic phonon mode red-shifted with respect to the bulk mode because of phonon confinement, and the latter is the l = 1 surface phonon. Analysis based on the phonon confinement model demonstrates that the size of the nanoparticle responsible for the red shift is about 4 nm, close to the estimate from the blue-shift of the photoluminescence. The Raman spectrum of ZnSe nanorods shows modes at 257 and 213 cm-1, assigned to longitudinal and transverse optic phonons, blue-shifted with respect to the bulk ZnSe modes because of compressive strain. The mode at 237 cm 1 is the surface phonon. PMID- 12908287 TI - Raman studies of semiconducting oxide nanobelts. AB - Crystalline nanobelts of ZnO and SnO2 were prepared from a thermal evaporation of oxide powders inside an alumina tube in the absence of catalysts. Typical dimensions of the nanobelt samples ranged from approximately 10 to 100 microns in length, 30 to 300 nm in width, and 6 to 30 nm in thickness. Room temperature Raman spectra were obtained on pressed mats of nanobelt samples and compared with the corresponding spectra of the starting oxide powders and bulk materials. Collectively, our Raman data indicated that the as-prepared nanobelt samples used in this study were oxygen deficient. Upon annealing at 900 degrees C in flowing oxygen for 1 h, the nanobelt samples exhibited Raman features that corresponded to those expected in respective bulk semiconducting oxides. The dimensions of the nanobelts were a bit too large to expect significant quantum size effects on the phonon structure similar to those observed in carbon nanotubes and short-period semiconductor superlattices. PMID- 12908288 TI - Bending properties of carbon nanotubes encapsulating solid nanowires. AB - Using empirical potentials and atomistic simulations, we model three-point bend tests of single-walled carbon nanotubes encapsulating metal nanowires. The presence of a metal nanowire inside the nanotube greatly suppresses the tube buckling instability. Increasing tube diameter leads to an increase in the bending strength; however, in contrast to hollow tubes, there is no decrease in the maximum deflection before buckling. Analysis of the principal bending vibrational mode shows a lowering of the frequency, associated with increased tube inertia. Remarkably, metal-filled tubes exhibit strong damping of oscillations whereas unfilled single-walled and multiwalled tubes show no damping. Our studies demonstrate the benefits of filling tubes with solids to modify bending strength and flexibility, suggesting applications for nanotube based elements in micromechanical devices or nanoprobes. PMID- 12908289 TI - Wet-chemical synthesis and properties of CoPt and CoPt3 alloy nanoparticles. AB - Surface-protected, air-stable nanoparticles of CoPt and CoPt3 were prepared by thermal decomposition/reduction of organometallic precursors with a long-chain aliphatic diol, also known as the polyol process. Particles 3 nm in diameter showed ferromagnetic behavior up to 350 K (Hc = 65 Oe at T = 300 K; Hc = 410 Oe at T = 5K) and underwent a disordering-ordering phase transformation after annealing that resulted in an increase in coercivity (Hc = 170 Oe at T = 300 K; Hc = 2000 Oe at T = 5 K). PMID- 12908290 TI - Mechanical alignment of quasi-one-dimensional nanoparticles. AB - Randomly oriented rod or rope-like nanoparticles on the surface of an elastomeric substrate are aligned along one direction simply by stretching the substrate. The technique is demonstrated here using single-walled carbon nanotube ropes, and the degree of alignment is assessed by polarized Raman spectroscopy. The alignment is preserved after the particles are removed from the substrate surface, showing that the aligned nanoparticles can be stamped in patterns onto another surface. PMID- 12908291 TI - Modeling of the HiPco process for carbon nanotube production. I. Chemical kinetics. AB - A chemical kinetic model is developed to help understand and optimize the production of single-walled carbon nanotubes via the high-pressure carbon monoxide (HiPco) process, which employs iron pentacarbonyl as the catalyst precursor and carbon monoxide as the carbon feedstock. The model separates the HiPco process into three steps, precursor decomposition, catalyst growth and evaporation, and carbon nanotube production resulting from the catalyst-enhanced disproportionation of carbon monoxide, known as the Boudouard reaction: 2 CO(g)- >C(s) + CO2(g). The resulting detailed model contains 971 species and 1948 chemical reactions. A second model with a reduced reaction set containing 14 species and 22 chemical reactions is developed on the basis of the detailed model and reproduces the chemistry of the major species. Results showing the parametric dependence of temperature, total pressure, and initial precursor partial pressures are presented, with comparison between the two models. The reduced model is more amenable to coupled reacting flow-field simulations, presented in the following article. PMID- 12908293 TI - Electromagnetic fields on a quantum scale. I. AB - This is the first in a series of two articles, the second of which provides an exact electro-magnetic field description of photon emission, absorption, and radiation pattern. Photon energy exchanges are analyzed and shown to be the triggered, regenerative response of a non-local eigenstate electron. This first article presents a model-based, hidden variable analysis of quantum theory that provides the statistical nature of wave functions. The analysis uses the equations of classical electro-magnetism and conservation of energy while modeling an eigenstate electron as a nonlocal entity. Essential to the analysis are physical properties that were discovered and analyzed only after the historical interpretation of quantum mechanics was established: electron non locality and the standing electro-magnetic energy that accompanies and encompasses an active, electrically small volume. The standing energy produces a driving radiation reaction force that, under certain circumstances, is many orders of magnitude larger than currently accepted values. These properties provide a sufficient basis for the Schrodinger equation as a descriptor of non relativistic eigenstate electrons in or near equilibrium. The uncertainty principle follows, as does the exclusion principle. The analysis leads to atomic stability and causality in the sense that the status of physical phenomena at any instant specifies the status an instant later. PMID- 12908292 TI - Modeling of the HiPco process for carbon nanotube production. II. Reactor-scale analysis. AB - The high-pressure carbon monoxide (HiPco) process, developed at Rice University, has been reported to produce single-walled carbon nanotubes from gas-phase reactions of iron carbonyl in carbon monoxide at high pressures (10-100 atm). Computational modeling is used here to develop an understanding of the HiPco process. A detailed kinetic model of the HiPco process that includes of the precursor, decomposition metal cluster formation and growth, and carbon nanotube growth was developed in the previous article (Part I). Decomposition of precursor molecules is necessary to initiate metal cluster formation. The metal clusters serve as catalysts for carbon nanotube growth. The diameter of metal clusters and number of atoms in these clusters are some of the essential information for predicting carbon nanotube formation and growth, which is then modeled by the Boudouard reaction with metal catalysts. Based on the detailed model simulations, a reduced kinetic model was also developed in Part I for use in reactor-scale flowfield calculations. Here this reduced kinetic model is integrated with a two dimensional axisymmetric reactor flow model to predict reactor performance. Carbon nanotube growth is examined with respect to several process variables (peripheral jet temperature, reactor pressure, and Fe(CO)5 concentration) with the use of the axisymmetric model, and the computed results are compared with existing experimental data. The model yields most of the qualitative trends observed in the experiments and helps to understanding the fundamental processes in HiPco carbon nanotube production. PMID- 12908294 TI - The exact electromagnetic field description of photon emission, absorption, and radiation pattern. II. AB - This is the second of two articles, the first of which contains a proposed explanation of quantum theory based upon electron nonlocality and classical electrodynamics. In this second article classical field theory is used to describe a unique field set for exchange of radiation between an atomic eigenstate and the far field. The radiation satisfies the thermodynamic condition of reversibility as described by Boltzmann, Planck, and Einstein. The exchanged radiation supports the kinematic properties of photons, and it can be emitted or absorbed by a vanishingly small volume. PMID- 12908295 TI - Chemical functionalization of carbon nanotubes. AB - There are many reasons why one would want to form chemical attachments to the walls or ends of carbon nanotubes: these chemical bonds might be used to tailor the interaction of the nanotube with other entities, such as a solvent, a polymer matrix, or other nanotubes. The chemically functionalized nanotube might have mechanical or electrical properties that are different from those of the unfunctionalized nanotube and thus might be used as a chemical sensor or a nanometer-scale electronic device. The challenge is to find a way to reproducibly and reliably chemically alter carbon nanotubes that, like graphite, are fairly unreactive. The various methods used to date and the possible application of the resulting functionalized nanotubes is discussed in this review paper. PMID- 12908296 TI - Synthesis of SnO2 nanoribbons by a carbothermal reduction process. AB - This communication describes, for the first time, the growth of SnO2 nanoribbons by a controlled carbothermal reduction process. An analysis of the transmission electron microscopy image revealed that these nanoribbons have a well-defined shape, with a typical width in the range of 70-300 nm. In general, the nanostructured ribbons were more than 100 microns in length. The results reported here support the hypothesis that this ribbon-like nanostructured material grows by a vapor-solid process. This study introduces two hypotheses to explain the SnO2 nanoribbon growth process. PMID- 12908297 TI - Nanoencapsulation of [60] fullerene by a novel sugar-based polymer. AB - [60] Fullerene has been nanoencapsulated by the formation of an inclusion complex with a beta-cyclodextrin-containing lactose copolymer. The cyclodextrin polymer was synthesized by reaction of beta-cyclodextrin chlorotriazine with beta lactose, and the polymer was highly soluble in water. Encapsulation of the carbon allotrope was achieved by forming a stable inclusion complex between the water soluble cyclodextrin and [60] fullerene by a new synthetic procedure. Preliminary studies indicate that this complex has potential for biomedical applications because of its radical scavenging properties. PMID- 12908298 TI - Surface modification of titanium carbide with carbyne-containing nanocoatings. AB - The aim of this research is to investigate a novel approach to surface engineering of biomaterials that are based on transition metals of the groups IVA VIA. The approach taken relies on the fact that, during the electropolishing of TiC surfaces, the removal of Ti atoms from the TiC surface surpasses that of C atoms. This leads to enrichment of the TiC surface with carbon. Transmission electron microscopic investigation showed that carbon-based films contain carbynes in the form of nanorod-like clusters with lengths in the range of 5-100 nm. This carbyne-containing layer is 50-100 nm thick. It was generalized that carbyne-containing nanofilms are formed on the carbide surface of transition metals of groups IVA-VIA during electropolishing. Since carbynes, being one dimensional chain-like structures [(-C identical to C-)n/(=C=C=)n] with sp1 carbon-carbon hybridization, have the highest degree of biocompatibility because of their biological activity, the development of such surface bioengineering with carbynes extends applications of biomaterials based on transition metals of the groups IVA-VIA. PMID- 12908299 TI - Variable-temperature scanning tunneling microscopy and scanning tunneling spectroscopy study on copper phthalocyanine ultrathin films on a Au(111) surface. AB - Variable-temperature high-resolution scanning tunneling microscopy (STM) images reveal that well-ordered copper phthalocyanine (CuPc) strips can be self assembled by depositing CuPc molecules on a Au(111) surface. The self-assembled strips are supposed to result from the balance of the intermolecular interaction and the interaction between the molecules and substrate during annealing. The energy band (approximately 1.9-2.1 eV) of CuPc, measured by scanning tunneling spectroscopy (STS), is comparable to the optical band gap (approximately 1.7 eV). Spectroscopic measurements confirm that a dipole layer and/or an effect of image force exist at the CuPc/Au(111) interface. PMID- 12908300 TI - Self-assembled nanocomposite of organic-inorganic hybrid: 2,4 dichlorophenoxyacetate in Zn-Al hydrotalcite-like layers. AB - Hydrotalcite-like inorganic layers of Zn-Al, a host containing an organic moiety, 2,4-dichlorophenoxy-acetate, as a guest, was prepared by the spontaneous self assembly method from an aqueous solution for the formation of a new layered organic-inorganic hybrid nanocomposite material. In this synthesis, the host- and guest-forming species were simultaneously included in the mother liquor, aged, and separated. Various Zn/Al ratios (R = 2, 3, and 4), concentrations of 2,4 dichlorophenoxyacetic acid (0.03-0.1 M), and pH (7 and 10) were studied to optimize the formation of the layered nancomposite. It was found that the optimum conditions for the formation of the nanocomposite were R = 4, pH 7, and concentration of 2,4-dichlorophenoxyacetic acid = 0.08 M. X-ray diffraction shows that this sample affords a nanolayered structure with a basal spacing of 24.6 A. PMID- 12908301 TI - Quasi-linear assemblies of silver nanoparticles by highly localized anodic dissolution of copper in the hydrosol. AB - The assembly of silver nanoparticles into quasi-linear superstructures in solution has been demonstrated. A small dc voltage applied to fine-tipped, L shaped copper electrodes immersed in aqueous carboxylic acid-derivatized silver nanoparticle solution leads to the anodic dissolution of copper ions. The controlled release of Cu2+ ions within the gaps results in local screening of repulsive coulombic interactions between the silver nanoparticles and the consequent formation of aggregates that are extremely stable in time. Atomic force and transmission electron microscopy images of the silver nanoparticle solution after application of an electric field showed the presence of a large number of silver nanoparticle assemblies in open, quasi-linear structures. Such open structures do not form if Cu2+ ions are added directly to the silver nanoparticle solution. PMID- 12908302 TI - Light-emitting devices based on ruthenium (II) bipyridine complexes coupled with cadmium sulfide nanoparticles. AB - The fabrication of a light-emitting device based on a composite consisting of cadmium sulfide nanoparticle, poly(vinylalcohol), and ruthenium 2,2'-bipyridyl complex is reported. The electroluminescence spectrum of this composite is similar to the combination of the emission from ruthenium complexes and cadmium sulfide. A stable electroluminescence at low threshold voltage (4 V), with a brightness of 680 cd/m2 and an external quantum efficiency of approximately 0.2% was achieved. it was demonstrated that the cadmium sulfide nanoparticles were able to improve the performance of the device and enhance the electron carrier mobility as compared to the device based on pure ruthenium complex. PMID- 12908303 TI - Effects of hydrogen on the formation of aligned carbon nanotubes by chemical vapor deposition. AB - Well-aligned carbon nanotubes with controllable properties were grown on porous silicon substrates by thermal chemical vapor deposition. The morphologies of the carbon nanotubes were varied with the introduction of H2 during the catalyst activation and/or carbon nanotube growth processes. It was found that H2 promotes the growth of carbon nanotubes while preventing the formation of spherical amorphous carbon particles. Without the introduction of H2 during the C2H2 thermal decomposition, aligned carbon nanotubes mixed with spherical carbon particles were formed on the substrate. However, with the introduction of H2, pure carbon nanotubes were synthesized. These nanotubes also had uniform diameters of 10-20 nm, which is much smaller than nanotubes synthesized without H2. The average growth rate of nanotubes was also affected by the introduction of hydrogen into the reaction chamber during nanotube growth. With the addition of hydrogen, the average growth rate changed from 78 nm/s to 145 nm/s. A possible growth mechanism, including the effect of a high ratio of H2 to C2H2, is suggested for the growth of these well-aligned carbon nanotubes with uniform diameters. PMID- 12908304 TI - Grain growth in nanocrystalline yttrium-stabilized zirconia thin films synthesized by spin coating of polymeric precursors. AB - This article reports results of experimental studies on the microstructural evolution of nanocrystalline yttrium-stabilized zirconia thin films synthesized on a Si substrate via a polymeric precursor spin-coating approach. Grain growth behavior has been investigated at different annealing temperatures (700-1200 degrees C) for periods of up to 240 h. A similar film thickness (approximately 120 nm) was maintained for all of the samples used in this study, to avoid variation in film thickness-dependent grain growth. The effects of the thermal history of the film and the annealing atmosphere on the grain growth were also studied. A simple semiempirical grain growth model has been developed to describe isothermal annealing data and to predict dynamic grain growth behavior during the sintering of polymeric precursor layers to form cubic-phase nanocrystalline yttrium-stabilized zirconia films. PMID- 12908305 TI - New poly(butylene succinate)/layered silicate nanocomposites: preparation and mechanical properties. AB - New poly(butylene succinate) (PBS)/layered silicate nanocomposites have been successfully prepared by simple melt extrusion of PBS and octadecylammonium modified montmorillonite (C18-mmt) at 150 degrees C. The d-spacing of both C18 mmt and intercalated nanocomposites was investigated by wide-angle X-ray diffraction analysis. Bright-field transmission electron microscopic study showed several stacked silicate layers with random orientation in the PBS matrix. The intercalated nanocomposites exhibited remarkable improvement of mechanical properties in both solid and melt states as compared with that of PBS matrix without clay. PMID- 12908306 TI - Optical properties of the synthetic nanocomposites SiO2/CdS/poly(styrene-co maleic anhydride) and SiO2/CdS/poly(styrene-co-maleimide). AB - Hybrid materials consisting of SiO2/CdS particles dispersed in poly(styrene-co maleic anhydride) and poly(styrene-co-maleimide) have been synthesized and characterized. The polymer nanocomposites were synthesised in situ in the presence of previously prepared inorganic fillers (SiO2/CdS). The nanocomposites were synthesized with the use of as-prepared or surface-modified SiO2/CdS fillers. For both types of nanocomposites, the optical properties were evaluated and the observation of size quantization effects in the optical spectra is discussed. In this context, the influence of the inorganic fillers and polymer matrices on the optical properties of the final nanocomposites was investigated. PMID- 12908307 TI - Structural and magnetic properties of evaporated nanostructured Fe/V multilayers. AB - The structural and magnetic properties of the evaporated Fe/V multilayers with a fixed V-layer thickness (tV = 1.5 nm) and variable Fe layer thicknesses (0.75 nm < or = tFe < or = 6 nm) have been studied by X-ray reflectivity and high-angle X ray diffraction, conversion-electron Mossbauer spectrometry, and vibrating sample magnetometry. The results show that multilayers are formed with a broad Fe/V interface and pure crystalline bcc-Fe layers in the center of the individual subsystems. The Fe spin orientation is aligned in the film plane in the individual centers as well in the interfacial regions. The interfacial anisotropy constant Ks was estimated to be equal to 0.04 mJ/m2. This parallel magnetic anisotropy is discussed in terms of reduced symmetry effects on the hybridized 3d states. PMID- 12908308 TI - Nanocrystalline sensor-grade Sn1-xInxO2 (0 < or = x < or = 0.2). AB - Nanocrystalline Sn1-xInxO2 (0 < or = x < or = 0.2) has been successfully prepared by a solution chemical route. High-resolution transmission electron microscopy studies show that the average grain size of Sn0.8In0.2O2 heated at 310 degrees C, 500 degrees C, and 800 degrees C for 12 h is about 3-4 nm, 5-6 nm, and 7-10 nm, respectively. The corresponding values for pure SnO2 are 3-4 nm, 7-10 nm, and 50 90 nm, respectively. Powder X-ray diffraction and electron diffraction studies confirm the existence of solid solution only in the nanocrystalline state (the average particle size is in the range of 5-10 nm) with the solubility limited to 20% of In2O3. Indium ions stabilize the nanocrystalline nature of Sn1-xInxO2 (0 < or = x < or = 0.2) and prevent the grain growth by entering the SnO2 lattice. The thermal characteristics of nanocrystalline Sn1-xInxO2 (0 < or = x < or = 0.2) investigated by thermogravimetric (TG) and differential thermal analysis (DTA) show that the solid solution decomposes at 820 degrees C into SnO2 and In2O3, which is accompanied by a rapid crystal growth. The electrical conductivity and activation energy of Sn1-xInxO2 (0 < or = x < or = 0.2) undergo significant changes when the average grain size is less than or equal to 2 x the Debye length, LD. PMID- 12908309 TI - Bimetallic catalyst for synthesizing quasi-aligned, well-graphitized multiwalled carbon nanotube bundles on a large scale by the catalytic chemical vapor deposition method. AB - An effective method of growth by catalytic chemical vapor deposition (CCVD) to get a large-scale yield of carbon nanotubes is reported. In this method, acetylene is decomposed catalytically over well-dispersed metal particles (Co-Fe and Co-Ni) embedded in commercially available zeolite at a lower temperature (600 700 degrees C). The two binary-metal catalysts (Co-Fe and Co-Ni) used are compared by scanning electron microscopy (SEM) and transmission electron microscopy (TEM). Crucial reaction parameters, such as reaction time, temperature, and the effect of purity of gas to obtain optimum production of the nanotubes, both qualitatively and quantitatively, are also reported. PMID- 12908310 TI - Fabrication and characterization of vertically aligned carbon nanotubes on silicon substrates using porous alumina nanotemplates. AB - An ethylene-air laminar diffusion flame successfully provided silicon substrates of anodic aluminum oxide (AAO) template with vertically oriented well-aligned carbon nanotubes. Field emission scanning electron microscopy (SEM) showed that open-tipped carbon nanotubes consisting of tube elements with the same length and diameter uniformly coated the template. High-resolution transmission electron microscopy (TEM) analyses revealed these nanotubes to be multiwalled carbon nanotubes, some well graphitized. It was found that cobalt catalyst particles, but not the porous aluminum templates, helped the growth of carbon nanotubes through graphitization and bonding of carbon nanotubes to the silicon substrates. PMID- 12908311 TI - Transport properties of nanosystems: viscosity of nanofluids confined in slit nanopores. AB - A fundamental nonequilibrium statistical mechanical approach due to Pozhar and Gubbins (PG) is used to study the Poiseuille flow and momentum transport in 20 model nanofluids confined in slit pores several molecular diameters in width. A simplified version of a general expression for the PG theoretical viscosity is applied to calculate the localized viscosity of the nanofluids in terms of the equilibrium structure factors (density and correlation functions) of nanosystems. These structure factors are calculated by means of the equilibrium molecular dynamics simulations. The localized theoretical viscosity so obtained is used further to calculate the theoretical pore-average viscosity of the nanosystems, and the latter is successfully compared with that extracted from nonequilibrium molecular dynamics simulation data. A simple correlation between the pore-average velocity, viscosity, nanofluid density, and the pore width for nanosystems of moderate density has been developed and recommended for applications in engineering. PMID- 12908313 TI - Organic-inorganic nanostructured colloids. AB - The synthesis and applications of organic-inorganic nanostructured colloids and colloidal-based materials are reviewed here. Emphasis is placed on the strategies and synthetic methods developed to organize organic-inorganic architectures. The article begins with a description and a general hierarchical classification of different systems, from inorganic particle synthesis and surface modification to more elaborate nanostructured colloids obtained through in situ encapsulation and/or self-assembly techniques. Ordering of colloids into two- and three dimensional arrays and their use as templates is also considered. For every system, typical examples are given that highlight the advantages and limitations of the techniques, as are more recent developments. Some properties and applicability of organic-inorganic colloids in catalysis, medicine, and coating technologies are also cited. PMID- 12908312 TI - Atomic force microscopy study of the interaction of Fusarium solani pisi cutinase with lipid surfaces. AB - We present an atomic force microscopy (AFM) study of a supported triacylglyceride multilayer phase and its interaction with lipolytic enzyme cutinase from Fusarium solani pisi. The multilayer triacylglyceride phase of coconut oil showed a rippled surface structure in the AFM images. Upon enzymatic degradation of the triacylglyceride phase, the ripple structure vanished rapidly. The apparent catalytic rate constants could be estimated based on the AFM image information. Interestingly, in one sample we observed what we interpret as a recurrent structural collapse of the cavity dug out by the protein. We interpret the cavities seen in the AFM images as molten surfaces or surface holes filled with liquidified phase containing product molecules, which appear transparent during the image recording. PMID- 12908314 TI - Preparation of graphitic carbon nanofibers with the use of water-soluble supports. AB - Graphitic carbon nanofibers (GCNFs) are prepared from CO/H2 with a nickel-rich Fe Ni (2:8) growth catalyst supported on three different water-soluble supports (Na4SiO4, Na2SiO3, and Na2CO3). GCNF products are formed with yield and crystallinity comparable to those of GCNF product produced with the same growth catalyst supported on fumed silica. Separation of GCNF product from the solid support is accomplished by aqueous dissolution of the support. This synthetic method extends the convenient isolation of GCNFs formed with supported growth catalysts to a wide variety of potential solid supports. PMID- 12908315 TI - Layer-by-layer nanostructured hybrid films of polyaniline and vanadium oxide. AB - Supramolecular structures of polyaniline (PANI) and vanadium oxide (V2O5) have been assembled via the electrostatic layer-by-layer (ELBL) technique. Strong ionic interactions and H-bonding impart unique features to the ELBL films, which are distinct from cast films obtained with the same materials. The interactions were manifested in UV-vis and Fourier transform infrared spectroscopy data. They are enhanced by the intimate contact between the components, as the films are molecularly thin, with 25 A per PANI/V2O5 bilayer. PMID- 12908316 TI - A perfect carbon nanotube with two closed ends. AB - A perfect carbon nanotube with two closed ends consisting of graphitized carbon was experimentally observed. The large carbon nanotube, with a diameter of 187 nm and a length of 1.2 microns, was synthesized by pyrolysis of iron(II) phthalocyanine (FePc) under an Ar/NH3 flow at 900 degrees C on a nickel substrate. The structure and composition of the nanotube were determined by high resolution transmission electron microscopy and electron energy loss spectroscopy. The carbon nanotube seems to grow spontaneously by some autocatalytic process. PMID- 12908317 TI - Atomic force microscopic observation of the molecular orientation in ultrathin films of alkanoic acid-derivatized porphyrins on a mica surface. AB - The observation of the molecular orientation of alkanoic acid-derivatized porphyrins in ultrathin films deposited on mica was carried out by atomic force microscopy. It was observed that the tetraacid derivative 5, 10, 15, 20-tetra(N 10-carboxydecyl-pyridinium-4-yl) porphyrin was arranged as a monolayer with the porphyrin macrocycle oriented coplanar to the mica surface. On the other hand, the diacid derivative protoporphyrin IX Zn(II) formed a bilayer with the hydrophobic part inside and the hydrophilic part in the periphery. Therefore, in this case, the porphyrin macrocycle is roughly perpendicular to the mica surface. PMID- 12908318 TI - Molecular recognition by wall-assembling-type nanocavity in aqueous media. AB - Steroid cyclophanes, each having a macrocyclic ring attached to four bile acid moieties via chiral lysine connectors, were synthesized, and the binding of the 2 naphthylphenylketone (guest) to the steroid cyclophanes in water was investigated. The circular dichroism spectra of the steroid cyclophane with cholic acid and L-lysine were significantly affected by the binding of the guest, and the induced circular dichroism based on the absorption of the achiral guest was also observed. The binding of the guest to the steroid cyclophane with cholic acids and D-lysines induced changes in the circular dichroism spectra with the opposite sign of the molecular ellipticities. An induced circular dichroism spectral change was not observed upon binding of the guest to the analogous host without OH sites. These results strongly suggest that the guest is conformationally fixed through hydrogen bonding between the carbonyl group of the guest and the steroidal hydroxyl group of the host. The assembly of only four steroid residues on the macrocyclic ring probably provided a hydrophobic nanocavity for hydrogen bonding. PMID- 12908319 TI - Nanocrystalline alpha-Ni(OH)2 prepared by ultrasonic precipitation. AB - Nanocrystalline alpha-Ni(OH)2 was prepared by an ultrasonic precipitation/stirring method. Results of X-ray diffraction, transmission electron microscopy, infrared, and thermogravimetric measurements confirm that the sample obtained is alpha phase. Compared with the sample prepared without ultrasonic stirring, the crystal structure of the alpha phase sample has been changed from beta phase. The crystalline size of the sample is about 20 nm, which is smaller than the sample produced without ultrasonic stirring (70 nm). PMID- 12908321 TI - Replication molds having nanometer-scale shape control fabricated by means of oxidation and etching. AB - A means of accurate control of the curvature radius of molds that are used in nanostructure replication techniques is presented. The local non-uniform growth of SiO2 at regions with high curvature is used to fabricate molds with a curvature radius ranging anywhere between 10 and 250 nm. The mold radius is predicted by numerical simulation as a function of oxidation temperature and time and confirmed by a series of oxidation and etching experiments. The silicon, silicon dioxide, and polymer nanostructures are analyzed by scanning electron microscopy and compared with the theory. Replication into photo-plastic polymer from various sharp and round molds is performed, and their properties are discussed. Our results are useful for designing nanostructures in the area of soft lithography and nanoprobe engineering. PMID- 12908320 TI - Voltage tunable electroluminescence of CdTe nanoparticle light-emitting diodes. AB - Strong electroluminescence was observed in a CdTe nanoparticle light-emitting diode that was fabricated by layer-by-layer assembly of poly(p-phenylenevinylene) and CdTe nanoparticles. The turn-on voltage for the electroluminescence is 4.0 V. The electroluminescence is exclusively from CdTe nanoparticles, with a 30 nm red shift in emission wavelength from the photoluminescence and shifts to longer wavelengths at higher applied voltages. Possible mechanisms for this red shift are discussed, and it is concluded that the de-trapping of surface states and trap centers due to electric excitation, the involvement of traps in the electroluminescence, and the quantum confined Stark effect are the major reasons for the red shift. PMID- 12908322 TI - Nanoparticles of latexes from commercial polystyrene. AB - Nanoparticles of polystyrene (PS) (Mw = 1.0-3.0 x 10(6) g/mol) latexes have been successfully prepared from their respective dilute PS (commercial) solutions in cyclohexane, toluene/methanol, or cyclohexane/toluene at each theta temperature. The cationic surfactant cetyltrimethylammonium bromide (CTAB) was used to stabilize the formed PS latex particles. By varying different concentrations of CTAB and PS solution of various Mw, we have successfully produced, for the first time, stable bluish-transparent latex particles ranging from about 10 to 30 nm in diameter (Dw). The number of polymer chains per latex particle (np) is directly proportional to the volume occupied by each latex particle and hence associated to its Dw. The characteristics of these preformed PS latex particles are quite similar to those obtained from the microemulsion polymerization of styrene as reported in literature. These PS latex particles could be further grown by seeding polymerization of styrene to about 50 nm (Dw) with a monodisperse size distribution of Dw/Dn = 1.08. PMID- 12908323 TI - Synthesis and characterization of lanthanum carbide nanotubes. AB - Lanthanum carbide nanotubes have been synthesized by d.c. arc evaporation (approximately 20-30 V, approximately 200 Amp) of lanthanum metal (90 wt.%) loaded graphite rod in a helium atmosphere (665 mbar). To explore the possibility of formation of lanthanum carbide nanotubes, the experiments were carried out with lanthanum metal in different concentrations (i.e., 30, 50, 70, and 90 wt.%) in the graphite rod. The as-synthesized samples were characterized by transmission electron microscopy and X-ray diffractometry. Lanthanum carbide nanotubes (LCNTs) with a diameter of approximately 65 to 95 nm and a length of approximately 0.2 to 1.5 microns were obtained in this study. PMID- 12908324 TI - Nanoindentation behavior of clay/poly(ethylene oxide) nanocomposites. AB - Depth-sensing nanoindentation of clay/poly(ethylene oxide) nanocomposites has shown significant differences in the mechanical behavior of the composites, depending on method of synthesis and clay content. In general, the clay/polymer nanocomposite approach can dramatically improve the hardness, stiffness, and creep resistance of the polymer. However, these improvements are dependent on the clay content. At lower concentrations, the improvements are not significant, because of an adverse effect on creep behavior. PMID- 12908325 TI - Synthesis and characterization of PtSn/carbon and Pt3Sn/carbon nanocomposites as methanol electrooxidation catalysts. AB - Two Pt-Sn/vulcan carbon nanocomposites containing nanoclusters of PtSn (niggliite) and Pt3Sn highly dispersed on a carbon powder support have been prepared using Pt(SnPh2Cl)(PPh3)2(Ph) or [Pt3[mu-(PPh2)2CH2]3(mu 3-SnF3) (mu 3 CO)][PF6] as single-source precursors of metal alloy. PtP2 or Pt metal is also present as a secondary phase. Bimetallic Pt-Sn nanoclusters with an average diameter of 5-8 nm are formed at a total metal loading of ca. 15 wt%. Evaluation of both Pt-Sn/C nanocomposites as electrooxidation catalysts in a direct methanol fuel cell gives fuel cell performances comparable to that expected for Pt-Sn catalysts prepared by more conventional methods. PMID- 12908326 TI - Synthesis of mesoporous silica with embedded nickel nanoparticles for catalyst applications. AB - Here we describe a new route for the synthesis of nanometric Ni particles embedded in a mesoporous silica material with excellent potential for catalytic applications. Mesoporous silica with a surface area in the range of 202-280 m2/g, with narrow pore size distribution and Ni nanoparticles (particles in the range of 3-41 nm) were obtained in a direct process. A different approach was adopted to process such a nanocomposite. This new approach is based on the formation of a polymer with the silicon oxianion and nickel cation chelated to the macromolecule structure and on the control of the pyrolysis step. The CO/CO2 atmosphere resulting from the pyrolysis of the organic material promotes the reduction of the Ni citrate. PMID- 12908327 TI - Silica-precipitating peptides isolated from a combinatorial phage display peptide library. AB - Many biological organisms contain specialized structures composed of inorganic materials. Cellular processes in vivo facilitate the organized assembly of mineral building blocks into complex structures. The structural hierarchy and complexity across a range of length scales are providing new ideas and concepts for materials chemistry. Proteins that direct biomineralization can be used to control the production of nanostructured materials and facilitate the fabrication of new structures. Here, we demonstrate that some of the silica-binding peptides isolated from a combinatorial phage peptide display library can be used in precipitating silica from a solution of silicic acid. The results described in this report demonstrate that peptides displayed by phages act as templates in inorganic material synthesis and provide a means of understanding how some of the biological systems may be carrying out materials chemistry in vivo. PMID- 12908328 TI - Observation of sulfur on the surface of giant aurothiol nanoclusters. AB - We report the study of aurothiol nanoclusters using high-resolution electron microscopy, energy loss spectroscopy, X-ray photoelectron spectroscopy, Auger spectroscopy, and microscopy. It is concluded that the sulfur atoms are located on the surface of the gold nanoparticles in both (100) and (111) microfacets. The X-ray photoelectron spectroscopy data show that there is a Au-Au bond as well as a Au-S bond. Auger depth profile measurements made by sputtering of the nanoparticles corroborates that the sulfur is located on the surface of the nanoparticle. Quantitative Auger analysis indicates a ratio Au/S between approximately 1.79 and 1.98. PMID- 12908329 TI - Synthesis of nanoparticles of barium lanthanum hafnium oxide by a modified combustion process. AB - Barium lanthanum hafnium oxide, a complex perovskite ceramic, has been synthesized as nanoparticles by a modified combustion process for the first time. The Ba, La, and Hf ions required for the formation of Ba2LaHfO5.5 were obtained in solution by dissolving in boiling nitric acid a stoichiometric mixture of BaCO3, La2O3, and HfO2 that had been heated at 1200 degrees C for 4 h. By complexing the ions with citric acid and using ammonia as fuel, it was possible to get Ba2LaHfO5.5 as nanoparticles in a single-step combustion process. The powder obtained by the present combustion process was characterized by X-ray diffraction, BET surface area analysis, differential thermal analysis, thermogravimetric analysis, infrared spectroscopy, and scanning and high resolution transmission electron microscopy. According to the results of X-ray and electron diffraction, the powder synthesized through the combustion process showed single-phase barium lanthanum hafnium oxide. The transmission electron microscopic investigations showed a grain size of 42 nm, with a standard deviation of 8 nm. The nanoparticles of Ba2LaHfO5.5 synthesized by the present combustion technique could be sintered to > 97% of the theoretical density at a relatively low temperature of 1425 degrees C. Scanning electron microscopic studies on the sintered Ba2LaHfO5.5 samples showed that the final grain size of the sintered specimen was < 500 nm. PMID- 12908330 TI - Changes in center-of-pressure dynamics during upright standing related to decreased balance control in young adults: fractional Brownian motion analysis. AB - We investigated the relationships between the ability to maintain balance in an upright stance and center-of-pressure (COP) dynamic properties in young adults. Included in this study were 10 healthy male subjects in each of two groups with respect to balance ability. Balance ability was evaluated according to the length of time a subject stood on one leg with his eyes closed. The means and ranges of this one-leg balancing time were 17.9 s (3-43 s) and 118.3 s (103-120 s) for the off-balance and balance groups, respectively. The time-varying displacements of the COP under a subject's feet during quiet two-leg (normal) standing were measured by an instrumented force platform. Each subject was tested in both the eyes-open and eyes-closed conditions. The COP trajectories were analyzed as fractional Brownian motions according to the procedure of 'stabilogram-diffusion analysis', proposed by Collins and De Luca (1993). The extracted parameters were the effective diffusion coefficients (D) for the short-term (less than about 1.0 s) and long-term intervals, respectively, as well as the Hurst exponents (H) for the short-term and long-term intervals, and some critical-point coordinates (i.e., critical mean square displacements and critical time intervals). The off balance group showed significantly higher values for short-term D, short-term H, and critical mean square displacements than the balance group. No significant differences between the groups were found in the long-term D and H or in the critical time intervals. That is, for the off-balance subjects, an increase in the stochastic activity and positively correlated (persistent) behavior of the postural sway during shorter timescales may cause postural instability. These results suggest that the difference in balance ability for young adults is related to the open-loop (i.e., short-term) control mechanisms but not to the corrective feedback (i.e., long-term) mechanisms used to maintain balance in an upright stance. PMID- 12908331 TI - Environmental, health and economic conditions perceived by 50 rural communities in Bangladesh. AB - For randomly selected 50 villages in Bangladesh, an interview survey with a structured questionnaire was conducted to reveal their perception on the environmental, health and economic conditions at present and for the past 10-year change. The eight following items were analyzed in this paper: air pollution and water pollution, which represent environmental conditions with close relation to health conditions, soil degradation and deforestation, which represent environmental conditions with close relation to economic conditions, epidemic diseases and malnutrition, which represent health conditions, and poverty and jobless, which represent economic conditions. Among the 50 villages, deforestation was most frequently perceived serious at present and worsened in the past 10 years. Of the remaining seven items, those related to economic conditions were more seriously perceived than those related to health and environmental conditions. As revealed by the cluster analysis for the inter-item relations, epidemic diseases, which formed the same cluster with the environmental items, were recognized less serious whereas malnutrition, which formed the same cluster with the economic items, was recognized more serious. These findings are useful not only for rural development programs but also for mitigation programs toward health and environmental hazards in Bangladesh. PMID- 12908332 TI - Kinetic study of seita-fitting. AB - We have already presented two studies of the traditional carrier frame, the seita. In our first study, we reported on seita users supporting loads not on the lumbar vertebrae but on the sacrum. In the second study, we showed that carrying a load on the sacrum was efficient in terms of metabolic rate, muscle activity, cadence and subjective responses. The purpose of this study was to verify the effect of carrying a load on the sacrum in terms of gait pattern. We compared the kinetic parameters produced while carrying a load on the sacrum (LOS) with those produced while carrying a load on the lumbar vertebrae (LOLV). Maximum propulsive force and medial impulse were significantly larger in LOS than in LOLV. These results suggested that a normal gait pattern was maintained more in LOS conditions than in LOLV conditions. This indicated that seita-fitting was efficient for carrying and transporting loads. PMID- 12908333 TI - Evaluation of a step-test for assessing the cardiorespiratory capacity of workers in Thailand: a pilot study. AB - The aim of this pilot study was to evaluate the accuracy of a simple step-test procedure supplemented by the measurement of heart rate (HR) (STEP1) to assess the cardiorespiratory capacity i.e., maximal oxygen consumption (VO2max) of Thai workers. The subjects comprised 18 men and 17 women. Their ages varied from 19 to 20 years and all were physically active. The subjects performed three tests: a submaximal incremental cycle-ergometer (CYCLE) test supplemented by HR recordings and respiratory gas exchange, and two step-tests (STEP1 and STEP2). In the STEP1 test HR was recorded continuously. The STEP2 test included the measurements of HR and respiratory gas exchange. For the male subjects the mean difference of the estimated VO2 max values obtained from the CYCLE and STEP1 test was 15% (p < 0.05). With the female subjects the mean VO2 max values were equal when predicted according to the CYCLE and STEP1 tests. The present STEP1 test is not sufficiently accurate to predict the VO2 max of Thai men. The results of this pilot study need to be confirmed with larger samples of subjects with various backgrounds in terms of individual characteristics and occupations. PMID- 12908334 TI - Fatigue complaints among female shift workers in a computer factory of Japan. AB - For female workers in a computer factory in Japan, consisting of 41 daytime workers and 74 weekly-rotating shift workers (of whom, 37 each were engaged in, respectively, early-shift work and late-shift work during the survey week), within-day variations in the number of fatigue complaints were elucidated. Based on a repeated questionnaire survey, changes of fatigue complaints in a day were evaluated at three occasions, i.e., just before work, just after work, and before retiring, for three working days and one off day. The occasions of fatigue feelings differed among the three work groups: the complaints were significantly more frequent before work in the early-shift workers, after work in the late shift workers, and before retiring in the daytime workers. Feeling of fatigue before and after work may be disadvantageous to safety and efficiency of work. PMID- 12908336 TI - Gender differences in memory for film of pigmies' hunting-gathering activities among Japanese children and adults. AB - Although adaptation to hunting-gathering life is a main hypothesis for understanding of the nature of humans, studies directly examining the hypothesis have not been done. In the present study, we used the method of showing a film depicting hunting and housework by African hunter-gatherers to elementary pupils and university students to examine their memories. In pupils and students, males showed higher percentage of correct answers than females for hunting-related questions, and female showed higher percentage for housework-related questions. The results suggest a males' learning bias to hunting and support the hunting gathering hypothesis. PMID- 12908335 TI - Recovery cardiac cost following short duration high intensity exercise in prepubertal, just pubertal, and postpubertal girls and its relationship with physical and physiological parameters. AB - Recovery cardiac cost (RCC) after short duration high intensity exercise and its relationship with physical and physiological parameters were assessed for 45 sedentary girls aged 10-25. RCC of postpubertal girls were significantly lower than those of prepubertal and just pubertal girls, when expressed in terms of cost in beats above rest. No significant differences were found among the three groups regarding RCC values in terms of % cost above rest. Age and diastolic blood pressure were negatively correlated with RCC both in terms of cost in beats above rest % cost above rest only in postpubertal group. Pre-exercise heart rate in post-pubertal group was negatively correlated with RCC only in terms of % cost above rest. No positive or negative correlation was found between RCC and other parameters under study, in prepubertal and just pubertal groups. PMID- 12908337 TI - [Prevalence of osteopenia and osteoporosis assessed by densitometry in postmenopausal women]. AB - OBJECTIVE: Our goal was to know the osteopenia and osteoporosis prevalence, determined by densitometry using dual energy X-ray absorptiometry (DXA) to measure the bone mineral density (BMD) in postmenopausal women. SUBJECTS AND METHODS: A total the 202 postmenopausal women aged 37 at 74 years old was studied, at the Gynecologist and Obstetrics Hospital No. 3 Medical Center "La Raza" from the IMSS, initiated in December 2001 to June 2002. The measurement BMD using DXA with a LUNAR device and according the WHO criteria, we have calculated the prevalence of osteopenia and osteoporosis in normal postmenopausal women at the lumbar spine (LS) and/or femoral neck (FN). RESULTS: The osteopenia prevalence in LS was: 18.2% in the group aged 45-49 years old, 47.1% in the group aged 50-54, 43.7% in the group aged 55-59, 63.6 in the group aged 60-64, 53% in the group aged 65-69, and 30% in the group aged 70-74. The overall osteopenia prevalence in LS was 43.5%, confidence interval (CI) 95% from 43.08 to 44.03%. The osteoporosis of LS was 17.1% in the group aged 50-54 years old; 21.8 in the group aged 55-59; 22.7 in the group aged 60-64; 35.29% in the group aged 65-69; and 30% in the group aged 70-74. The overall osteoporosis prevalence in the LS was 19.8% (CI) 95% from 14.3 to 25.2%. The osteopenia prevalence in FN was. 19% in the group aged 45-49 years old; 40% in the group aged 50-54; 49% in the group aged 55-59; 54% in the group aged 60-64; 76% in the group aged 65-69 and 60% in the group aged 70-74. The overall osteopenia prevalence of FN was the 48% CI 95% from 40.65-54.59%. The osteoporosis prevalence in FN: 2.38% in the group aged 45 49 years old; 7.14 in the group aged 50-54; 5.45% in the group aged 55-59; 13.6% in the group aged 60-64; 5.8 in the group aged 65-69, and 20% in the group aged 70-74. The overall osteoporosis prevalence in FN was 7.4% IC 95% from 3.6-10.68%. CONCLUSIONS: The osteoporosis prevalence in this group is less than between American women (15%), and early detection is possible between women with BMD low, in the postmenopausal women with high risk factors and primary treatment need to be established and implemented. PMID- 12908338 TI - [Frequency of altered male factor in an infertility clinic]. AB - Infertility is the incapacity of a couple to conceive after a year of regular sexual life without using a method for family planning. The infertility state is dependent on the female factor as well as masculine factor; an altered masculine factor is designated when any cause or causes of infertility reside in the male. The masculine factor as a cause of infertility is present in 40 to 50% of cases hence the importance of an integral evaluation of male alterations and its fertility. The World Health Organization (WHO) has proposed to classify the masculine infertility based on semen features. OBJECTIVE: To determine the frequency of the masculine factor altered in association to diagnosed abnormalities, according to the number of affected seminal indexes based on seminograms performed in infertile patients seen at the department of Human Reproductive Medicine, Hospital Juarez de Mexico. MATERIAL AND METHODS: An observational, descriptive, transverse, retrospective study was performed. SUBJECTS: the seminograms practiced were reviewed from 571 clinical files of couples that the consultation for infertility from January 1993 to February 2001. Collection and analysis of semen samples was based on standards settled by WHO. RESULTS: Of 571 seminogram informs, 371 (65%) showed alterations in the seminal indexes, the stocking of age of this group was of 31.89 years +/- SD 6.3 years. Our findings were: azoospermia in 89 (23.98%), astenonecrozoospermia in 44 (11.85%), hypospermia in 43 (11.59%), astenozoospermia in 33 (8.89%), oligoastenozoospermia in 31 (8.35%) hyponecrozoospermia in 30 (8.08%), oligozoospermia in 25 (6.73%), others in 21 (5.7%), necrozoospermia in 19 (5.12%), hypoastenozoospermia in 18 (4.85%), cryptozoospermia in 11 (2.97%), and hypooligoastenozoospermia in 7 (1.88%). In 62.6% of seminograms practiced there was an isolated alteration, in 34.4% two alterations and in the remaining 3% more than two alterations. CONCLUSION: We found masculine factor altered in 65% of couples who were requiring consultation for infertility. The three main alterations of seminal indexes were azoospermia, astenonecrozoospermia and hypospermia. PMID- 12908339 TI - [Early hebdomadal complications in newborns from mothers with both mild and severe preecampsia]. AB - OBJECTIVE: To understand the early hebdomadal complications in newborns from mothers with both mild and severe preeclampsia. MATERIAL AND METHODS: A transversal, observational trial was conducted in HGR No 1 IMSS, Morelia, Michoacan, Mexico. The clinical records of gestating women were reviewed to detect those who had preeclampsia (PE) or eclampsia in the tokosurgical ward of the same hospital. Newborns were sent up to neonatology room, where their perinatal clinical history was done, and were also observed to detect complications. RESULTS: During the trial 1,644 gestating women were attended: 49 (3%) had PE, 25 (51%) of which had mild PE, 24 (49%) severe PE. Gestational age in 12 (25%) was less at the 37 weeks. With regard to delivery resolution, cesarian section was performed in 28 (57%). As to neonates, one or more early hebdomadale complications were observed in 32 (65%), in which asphyxia, hypoglucemia, and hypocalcemia as well as necrotizing enterocolitis predominated. Multivariate and univariate analysis showed severe preeclampsia to be associated as a significant and independent risk factor for asphyxia, hypocalcemia and necrotizing enterocolitis (RR 7.2, CI 95% 1.8-2.8; RR 1.3 CI 95% 1.0-1.7; and RR 1.4 CI 95% 1.0-1.9 respectively) (p < 0.05). CONCLUSIONS: Disseminating the prenatal control significance to detect PE is important because of the repercussions over gestating women and fetus, which increases perinatal morbidity and mortality. PMID- 12908340 TI - [Association between insulin resistance and pregnancy induced hypertension. Case control study]. AB - OBJECTIVE: To evaluate the association between insulin resistance and the pregnancy induced hypertension. METHODS: One hundred sixty pregnant patients were studied in the third trimester: 46 patients with preeclampsia, 34 patients with gestational hypertension and 80 healthy patients (controls). Socio demographic data were collected and the body mass index was calculated as well as the insulin resistance was evaluated through the homeostasis model assessment. Three logistic regression analyses were made: First with all the patients who developed any form of pregnancy induced hypertension, the second with patients who developed gestational hypertension and the third regression with patients who developed preeclampsia. RESULTS: The variables that had statistical significance (P < 0.05) were number of pregnancies, parity, familiar incomes, and body mass index. The insulin resistance did not have statistical significance in any of the logistic regression analyses. CONCLUSION: The insulin resistance assessed in the third trimester of the pregnancy did not have association with the pregnancy induced hypertension and we recommend to carry out further investigations with prospective design and assessing insulin resistance calculating the homeostasis model assessment during all the pregnancy to evaluate this possible association. PMID- 12908342 TI - [Zoapatle's oxytocic action. 1948]. PMID- 12908341 TI - [Polycystic ovarian disease: clinical and biochemical expression]. AB - The policystic ovary (PO) results from a systemic hormonal dysfunction, characterized by hyperandrogenemia, insulin resistance and anovulation. OBJECTIVE: Assessment was made of clinical and biochemical features of patients with PO. MATERIAL AND METHODS: A retrospective analysis of 211 clinical of charts of patients with ultrasonographic diagnosis of PO seen at the Hospital Juarez de Mexico from 1996 to 2000 was performed. Evaluation was made of body mass index (BMI), waist hip index (WHI), Ferriman index, acanthosis nigricans, lipid and hormonal profile. RESULTS: Of patients studied (n = 211) 64% had infertility and abnormal menses associated to PO, while the rest was normal. Both groups (PO and normal), were similar in age and distribution as well as family back grown for diabetes mellitus, hypertensive disease and obesity. The clinical manifestations observed on patients with PO were infertility 79.16%, anovulation 68.42%, hirsutism 66.9% obesity (66.9%). Biochemical findings in PO patients vs normal patients were: hyperandrogenism (86.73% vs 71.4%), insulin resistance (60.46% vs 37.5%), dislipidemic (56.41% vs 31.5%), LH/FSH ratio > 2 (19.51% vs 10.51%). Regarding sensitivity and specificity for clinical and biochemical findings ranged from 0.64 at 0.90, minimum value had infertility and maximum anovulation; specificity (interval 0.31 to the 0.71) minor corresponded to hyperprolactinemia and higher to infertility. The estimated risk factor (OR) for the population studied exhibited anovulation (OR, 7; 95% IC, 1.79-32.92); infertility (OR, 4.51; 95% IC, 2.03-10.13); insulin resistance (OR, 3; 95% IC, 0.82-8.16); hyperandrogenism (OR, 2.61; 95% IC, 1.02-6.69); and obesity (OR, 2.16; 95% IC, 1.17-4). We concluded that population with PO has a higher risk of infertility, anovulation, obesity, hyperandrogenism, dyslipidemia, insulin resistance and abnormal menses. PMID- 12908343 TI - [Pleural fluid in chronic pancreatitis]. AB - In three patients, a 44-year-old schizophrenic woman and two men aged 54 and 42, who presented with dyspnoea, a pancreaticopleural fistula was diagnosed as a complication of pancreatitis, i.e. a fistulous tract between the pancreas and the pleural cavity. In general, these fistulas have a good prognosis; however, delay in finding the correct diagnosis influences the prognosis. This is often due to unfamiliarity with the disease and the non-specific presentation of patients with pleural effusion. The initial presentation usually comprises respiratory (dyspnoea, coughing due to pleural effusion) and occasional abdominal symptoms (epigastric pain). A definitive diagnosis is made when elevated pleural effusion amylase levels are demonstrated. Surgical treatment is only indicated if conservative or endoscopic treatment fails, and consists of resection of the fistula and drainage of the pancreatic duct via a lateral pancreaticojejunostomy or resection of the part of the pancreas where the fistula originates. In the first patient, surgical drainage of the fluid accumulation was applied, but she died of aspiration pneumonia after she had removed the feeding tube and had refused further treatment. Pancreatic resection resulted in recovery in the two men. PMID- 12908344 TI - [The prioritisation of genetic screening with primary haemochromatosis as an example]. AB - In 1994, the Health Council of the Netherlands published a report entitled 'Genetic screening' which contained 12 criteria for genetic screening programmes. However, the list does not prioritize the various criteria. From the list we have selected two criteria that we consider to be the most important. Firstly, the genetic screening test should be able to discriminate between subjects who are likely to develop the disease and those who are not. Secondly, there should be an effective treatment for subjects with the genetic defect. From this point of view, for example, screening for the C282Y mutation is not a suitable approach for detecting primary haemochromatosis. Although 85-90% of the patients with this disease are homozygous for this mutation, the majority of the carriers will not develop the disease. The 12 criteria of the Health Council of the Netherlands are still applicable. However, when taking a decision as to whether or not genetic screening is useful, we recommend that priority be given to the two primary criteria. PMID- 12908345 TI - [The use of apolipoprotein B in clinical practice to determine the risk for atherosclerosis]. AB - Atherosclerosis is a multifactorial disorder in which lipid profiles play an important role. In addition to the conventional lipid parameters such as total cholesterol, LDL cholesterol, HDL cholesterol and triglycerides, there is an increasing interest in plasma apolipoprotein B (apoB) for risk stratification in daily practice. ApoB is the structural protein of all atherogenic lipoproteins. Each of these lipoproteins contains one apoB molecule and therefore the apoB concentration is a direct reflection of the number of atherogenic particles in the blood. Some patients have a reasonably normal lipid spectrum but an elevated concentration of apoB, which is associated with an increased risk for atherosclerotic complications. This is especially the case for patients with atherogenic forms of hypertriglyceridaemia and with insulin resistance syndrome. On the basis of epidemiological data, it appears that apoB not only has added value as a risk parameter but also as a treatment target. However the clinical use of apoB levels requires a standardised method with better-defined reference values, as is already the case for total-cholesterol and LDL-cholesterol levels. PMID- 12908346 TI - ['Severe acute respiratory syndrome' (SARS): epidemiology, clinical signs, diagnosis and prevention]. AB - Severe acute respiratory syndrome (SARS) is caused by a recently identified Coronavirus (SARS-CoV). The clinical symptoms are non-specific and during the first few days in particular, are not clinically distinguishable from those of many other viral or bacterial infections. The majority of infected patients develop pneumonia within a week of the first symptoms appearing. Since November 2002 the virus has spread from South China to almost 30 other countries, where about 8500 infected individuals have been registered; about 800 people have already died from the disease (9.5%). The number of infected persons includes a noticeably high percentage of health workers. This fact underlines the importance of good infection prevention measures for each patient contact. The implementation of hygienic measures requires attention, because the infection of personnel in Toronto hospitals still occurred after the virus and transmission routes were known. It appears that transmission can be prevented with relatively simple precautions, as long as these are consistently implemented. Early recognition and isolation of a possible source are an essential part of this. SARS is a group A notifiable disease (report if suspected). In the Netherlands the general practitioner has a prominent role in assessing and treating individuals who are infected or might be infected with SARS-CoV. A protocol and a detailed action plan are available. In addition to this hospitals should be prepared for the initial reception of a patient with SARS, who presents directly to the outpatients' clinic or Casualty Department. PMID- 12908347 TI - [Diagnostic image (149). A woman with a mass at the base of the tongue. Lingual thyroid]. AB - A 48-year-old woman had a lump in her throat and dysphagia. On examination an oropharyngeal mass was seen which was diagnosed as a lingual thyroid. PMID- 12908348 TI - [Prediction of cardiovascular disease on the basis of blood pressure measurements more reliable with systolic than with diastolic measurement]. AB - OBJECTIVE: To find an answer in the literature to the question of which blood pressure parameter (systolic, diastolic or pulse pressure) best predicts cardiovascular disease and whether the magnitude of the association of the parameter varies with age. DESIGN: Structured literature review. METHOD: Pubmed was searched for prospective cohort studies concerning the predictive power of two or more blood-pressure components (in particular pulse pressure). RESULTS: Seven relevant articles were found. Systolic blood pressure was found to be a powerful predictor for cardiovascular disease at every age, while pulse pressure was found to be the weakest predictor. The predictive power of diastolic pressure was intermediate, and decreased with age whereas that of pulse pressure increased with age. CONCLUSION: Systolic blood pressure was the best predictor for cardiovascular disease, whereas the role of diastolic pressure appeared to be limited. PMID- 12908349 TI - [Drug treatment of chronic myeloid leukemia: a cost effectiveness analysis of first- and second-line treatment ]. AB - OBJECTIVE: To determine the average cost-effectiveness ratio of treatment of patients with chronic myeloid leukaemia (CML): with first-line interferon alpha 2a (IFN) or with second-line imatinib following IFN failure. DESIGN: Cost effectiveness analysis. METHOD: A general cost-effectiveness analysis was performed using a model. This model consists of two phases: an induction phase of eight months, in which newly-diagnosed patients are treated with either IFN or imatinib, and a chronic treatment phase wherein patients are treated according the result of the induction phase. The model calculated the costs and effects of the treatment. Input for this model was derived from literature and expert opinion. Costs were based on real cost prices and tariffs. RESULTS: Treatment with imatinib resulted in 6.67 quality-adjusted life years (QALYs) and treatment with IFN resulted in 4.98 QALYs. Average costs of treatment with 5 million IU/day of IFN were [symbol: see text] 76,969 and [symbol: see text] 53,257 with 3 million IU/day. For imatinib at 400 mg/day the costs were [symbol: see text] 140,765 per patient. Costs per QALY were [symbol: see text] 15,445, [symbol: see text] 10,687 and [symbol: see text] 21,082, respectively. CONCLUSION: The addition of imatinib to the treatment options in CML resulted in increased quality-adjusted survival, but also higher costs of treatment. PMID- 12908350 TI - [Four infants with congenital cystic malformation of the lung: different routes of diagnosis and forms of management]. AB - In four neonates, two boys and two girls, congenital cystic malformation of the lung was diagnosed. Two cases were diagnosed during prenatal ultrasound investigation. One of the patients recovered by a wait-and-see policy, one after antibiotic treatment and two following surgical removal of the diseased lung sections. Congenital pulmonary cysts are rare; congenital cystadenomatoid malformation (CCAM) is the most prevalent form. The most common presentation of CCAM is respiratory distress immediately following birth. Some children are asymptomatic at birth, but present with recurrent respiratory infections later in childhood. With the increasing use of antenatal ultrasound, an increasing number of cystic lung lesions are diagnosed antenatally. The prognosis of these mostly asymptomatic lesions is not clear and indications for prenatal and postnatal interventions must be determined by a multidisciplinary approach. Recently, malignant pulmonary tumours were found to be associated with CCAM. This finding necessitates long-term follow-up, even of asymptomatic CCAM lesions. PMID- 12908351 TI - [Meningitis due to Listeria monocytogenes as a complication of infliximab therapy]. AB - A 41-year-old woman was admitted to the hospital with meningitis caused by Listeria monocytogenes. Because of her Crohn's disease she used prednisolone and azathioprine. Two weeks before presenting with meningitis, infliximab had been given as the other immunosuppressant drugs had no effect. This tumour necrosis factor alpha (TNF alpha) blocking agent is known to increase the risk of opportunistic infections. This is the first Dutch patient described with meningitis caused by L. monocytogenes after treatment with infliximab. She recovered after antibiotic therapy. When antibiotic treatment is chosen, the possibility of opportunistic infections in patients who use infliximab concurrently with other immunosuppressant drugs should be taken into account. PMID- 12908352 TI - [Brain abscess: a difficult diagnosis]. PMID- 12908354 TI - [Increasing drug costs]. PMID- 12908353 TI - [Diagnostic image (140) A woman suffering from leg thrombosis]. PMID- 12908355 TI - [Increasing drug costs. The society's drug expenditures are strongly increasing. If this trend continues we have to make other priorities]. PMID- 12908356 TI - [Post-traumatic syringomyelia--a serious diagnostic suggestion?]. PMID- 12908357 TI - [Post-traumatic syringomyelia]. AB - INTRODUCTION: A longitudinal cavity in the medulla spinalis is known as a syringomyelia. If the cavity is a widening of the central canal, it is known as hydromyelia. Traumatic spinal cord injury may lead to the development of syringomyelia. MATERIAL AND METHODS: During the period 1959-1990 twenty patients with spinal cord injury in East Denmark (including Greenland) developed later symptomatic posttraumatic syringomyelia. RESULTS: The mean time from trauma to diagnosis was 12 years. Mean time from first symptom to time of diagnosis was three years. Thirteen patients were operated with syringoperitoneal drain, myelotomy or decompression of the spinal cord and dural reconstruction. The best effect of the operations was on pain; little or no effect was found on activities of daily living, motor function, spasticity, sensibility, and bladder and bowel function. DISCUSSION: The main objective of performing operation is to stop further development of disabling symptoms. We recommend that patients with spinal cord injury have an MRI performed three months after injury independent of recovery. If a cyst or syrinx is present or relevant clinical symptoms emerge, MRI should be performed every three months. If no cyst or syrinx or changes in neurological status develop, MRI should be performed every six months. If no changes are seen in two years, MRI should be performed every two or more years. PMID- 12908359 TI - [The prevalence of diabetes in Denmark. Development of a method for a registry based assessment]. AB - INTRODUCTION: The prevalence of diabetes in Denmark is unknown. The purpose of the present article was to describe the possibilities of developing a method to identify individuals with diagnosed diabetes in Denmark on the basis of existing national registers. MATERIAL AND METHODS: Record linkage of data from The Danish National Hospital Register, The Danish National Health Service Register and The Register of Medicinal Product Statistics. RESULTS: A minimum of 130,000 individuals in Denmark are estimated to have diabetes as on December 31, 1999. DISCUSSION: The possibility of developing a monitoring programme from existing data in registers seems to be present. If a validated monitoring programme is developed, this can not only be used to estimate the prevalence of diabetes, but also for a continuous monitoring of diabetes-related services. PMID- 12908358 TI - [Follow-up examination of Danish stainless steel welders previously examined in 1987]. AB - INTRODUCTION: A Danish cohort from 1987 consisting of 226 stainless steel welders and reference persons is part of the European Study Group on Cytogenetic Biomarkers and Health (ESCH). In ESCH increased cancer morbidity and mortality was significantly associated with high levels of chromosomal aberrations, measured in blood samples several years prior to cancer registration. The positive association was found in two cohorts from the Nordic countries and from Italy. MATERIAL AND METHODS: ESCH followed all registered cancer cases and control persons by questionnaires and interviews to obtain information about exposures in the period from the time of blood sampling for chromosomal aberration analysis to the time of cancer diagnosis. In Denmark the total cohort was included in the inquiry and the ESCH questions were supplemented with questions from the Danish National Work Environment Cohort Study 1990-95. RESULTS: Responses from one hundred and forty-four persons showed that seventy four were employed at the same workplace place as in 1987. Differences in occupational exposures, such as more noise, heat and insufficient lighting and no differences in the self-rated health were found in comparison with the Danish National Work Environment Cohort Study as such and with the sample of metal workers. Only very few of the study persons knew the threshold limit value of welding fumes but a majority found that the working environment had improved during the past ten years. DISCUSSION: This study confirms hazardous exposures in stainless steel welding. The threshold limit value, however, has been lowered since 1987 suggesting there is less cancer risk today from stainless steel welding. PMID- 12908360 TI - [Blockade of endothelin receptor with bosentan--a new principle of treatment in pulmonary arterial hypertension]. AB - Blockade of endothelin receptors is a new drug principle for the treatment of pulmonary arterial hypertension. Bosentan is a competitive blocker of endothelin receptors administered by the oral route. In clinical trials it has been shown to increase the functional capacity of patients with pulmonary arterial hypertension. This article describes the background, results of clinical trials, and practical guidelines for the use of bosentan in pulmonary arterial hypertension. PMID- 12908361 TI - [Uterine rupture during second trimester abortion induced with misoprostol]. AB - A 27-year-old woman with one prior cesarean section and one vaginal delivery was seen with a request for termination of pregnancy in her 17th week of pregnancy. Using vaginal misoprostol for two days the vaginal delivery failed. The fetus was giving way of previous scar resulting in rupture of the uterus, loss of blood and suture of uterus by laparotomy. PMID- 12908362 TI - [Long-term prophylaxis after venous thromboembolism]. PMID- 12908363 TI - [Drug therapy of Alzheimer dementia]. PMID- 12908364 TI - [130 kilometers/hour save time!--Besides there is a shortage of organ donors]. PMID- 12908365 TI - [Gatekeeper in the complex health care]. PMID- 12908366 TI - [Mentally ill Turks]. PMID- 12908367 TI - [When physicians cry]. PMID- 12908368 TI - [Are the integrators and supplements useful in sports?]. AB - The continued controversy regarding doping in Italian soccer turns the discussion to the use of food integration in sports. In Italy, there is only the word "integrator" to name many products so different regarding biological features and indication usage. We think it is useful, first of all, to talk about the meaning of integration, and then to introduce the differences between nutritional ergogenic aids and supplements and, finally, to think through relationship between nutrition and athletic performance. On the basis of recent literature, there is no evidence of any real benefit for athletes in taking nutritional ergogenic aids or supplements. PMID- 12908369 TI - [Cardiovascular effects of selective estrogen receptor modulators. Current perspectives]. AB - The use of hormone replacement therapy (HRT) after the menopause for the prevention of the long-term complications of estrogen deprivation has recently been questioned after the publication of large clinical trials that failed to show benefits for postmenopausal women. Although these trials risk to dump the widespread opinion of the cardioprotective effects of long-term estrogen use, they have many pitfalls that prevent a direct clinical application of these negative results. Furthermore, the large amount of epidemiological and experimental evidence indicating estrogens as protective on the vascular system cannot be ignored, and efforts should be devoted to understand the reasons for the discrepancy of results of these recent large trials. In the meanwhile, different molecules should be studied in depth as for the actions on the cardiovascular system, and their specific mechanisms of actions should be elucidated. Selective estrogen receptor modulators (SERM) are a promising family of molecules and some of these compounds have positive effects on cardiovascular risk parameters as well as on vascular cells. Large trials are ongoing to study the impact of these substances on cardiovascular risk, and the near future should provide us with answers on the possible use of SERM as possible safer alternatives to HRT for the long-term prevention of cardiovascular disease in postmenopausal women. PMID- 12908370 TI - [General practitioner and respiratory medicine specialist. A survey on the management of the asthmatic patient]. AB - The relationship between the general practitioner (GP) and the respiratory medicine physician specialist in the management of the asthmatic patients was examined. Two-hundred sixty -two GPs from Brescia and province were enrolled into the study and underwent an appropriate questionnaire, focusing the survey on three aspects: the relationship among GP-respiratory medicine physician specialist and asthma patients, the training of GP, the appraisal of the socio economic aspects of the management of bronchial asthma. The results have shown the requirement of a GP-respiratory medicine physician specialist relationship founded on the exchange of information and experiences, rather then of synthetic diagnoses and therapeutic directions, and the opportunity to establish outpatients' departments dedicated to the management of the asthmatic patients. Moreover, it has been shown an inadequate application of international guidelines and a not full appraisal of the socio-economic aspects of the management of the asthmatic patients. PMID- 12908371 TI - [Malnutrition and dementia: importance of care during food intake]. AB - Aim of this work is to assess the effects on nutritional status of appropriate care in food intake in demented patients. In a long term care in Turin, we enrolled 96 patients aged 75-103 years free from metabolic, hematologic and neoplastic diseases and not showing signs of acute illness. Seventy seven of these patients were affected by dementia. No statistically significant differences were found between demented patients and controls in nutritional status and daily nutrient intake and only need for care in food intake was independently associated with dementia. Our data demonstrate that similar caloric intakes can be obtained in demented and non demented patients with an appropriate level of care in food intake. PMID- 12908372 TI - [Primary plasma cell leukemia. A fulminant case]. AB - Primary plasma cell leukemia is a rare variant of malignant monoclonal gammapathies with de novo presentation in the leukemic phase. Only few series enrolling more than 20 patients have been reported in the literature. The authors deal with a patient who had a fulminant presentation of primary plasma cell leukemia with fatal outcome. Diagnosis of primary plasma cell leukemia was established based on Kyle's criteria, which include an absolute plasma cell count greater than 2 x 10(9)/L or a relative plasma cell number comprising greater than 20% of peripheral blood cells. The patient died within three days of diagnosis because of Streptococcus pneumoniae septic shock and multiple organ failure. This case report is worth of note due to the rarity of primary plasma cell leukemia and also because fulminant presentation is not commonly recognized among patients with malignant gammapathies. PMID- 12908373 TI - [HIV protease inhibitors for the treatment of Kaposi's sarcoma]. AB - A reduced incidence or the regression of Kaposi's sarcoma (KS) has been described in HIV-infected patients treated with HIV-protease inhibitors (PI). We have recently demonstrated that PI block the angiogenesis and the development of KS lesions induced experimentally in vivo by the inoculation of angiogenic factors or human primary KS cells. These effects of PI occur at the same drug concentrations in plasma of treated individuals, and they are due to the inhibition of cell invasion and of the activation of matrix metalloprotease-2, an enzyme that is key to angiogenesis and tumor growth and invasion. Since PI also block the production of cytokines involved in KS initiation and maintenance, this anti-inflammatory activity of PI may also contribute to the anti-KS effects observed in treated individuals. Thus, by direct and indirect activities PI can simultaneously block several pathways involved in tumor growth, invasion or metastasis. These data indicate that PI should also be investigated and exploited for the therapy of KS and tumors of different histology occurring in non infected individuals. PMID- 12908374 TI - [Celiac disease and irritable bowel syndrome: association or diagnostic error?]. AB - Some recent studies showed that coeliac disease is diagnosed in 6-30% of patients previously classified as affected by irritable bowel syndrome. We described herein which are the arguments pro and cons the association between coeliac disease and irritable bowel syndrome, and which is the correct diagnostic algorithm to perform the correct diagnosis. PMID- 12908376 TI - Anemia in patients with coronary artery disease. AB - The pathophysiology of impaired hemodynamic and nonhemodynamic responses to anemia in patients with coronary artery disease is discussed. In animals, experimentally induced coronary artery disease significantly inhibits the hemodynamic response to surgical blood loss; anecdotal evidence in humans corroborates these findings. Erythropoietic response to surgical blood loss may also be blunted in patients with coronary artery disease. Regardless of whether anemia is the result of a preexisting condition or surgical blood loss, its presence worsens outcomes in patients with coronary artery disease who undergo cardiac surgery. The combination of coronary artery disease and anemia has resulted in acute myocardial infarction as well. Finally, anemia after noncardiac surgery is associated with an increased risk of myocardial ischemia, potentially creating a cycle in which blood loss and myocardial ischemia exacerbate each other. Oral iron replacement therapy after elective cardiac surgery increases adverse events without significantly improving hematocrit and hemoglobin levels or iron stores. Allogeneic blood transfusions are less than ideal, and autologous blood transfusion with erythropoietin administration is only possible before elective procedures. New procedures and medications have reduced the blood loss associated with percutaneous coronary intervention, and minimization of blood loss during percutaneous coronary intervention has potentially major clinical and economic implications. PMID- 12908375 TI - [Asthma and aspirin]. AB - Aspirin (ASA) is an important cause of asthma so that ASA induced asthma (AIA) is considered a disease. Its prevalence is of 0.3-0.6 in the general population but it raises to 21% in the asthmatic one. Middle aged female are the most affected. AIA generally begins with a non allergic rhinitis, complicated sometimes with polyps, that evolves secondarily in asthma. The disease is often so severe to need oral corticosteroids to be controlled. It persists independently to the intake of ASA. From a pathogenetic point of view the interaction of ASA with the arachidonic acid metabolism seems to be important. The inhibition of cyclo oxygenase (COX) induces an activation of lypo-oxygenase with an increased synthesis of leukotrienes. In ASA intolerant patients there is also an activation of LTC4 synthetase, enzyme responsible for the synthesis of leukotrienes. Clinical history is very important to diagnose the disease but to confirm the diagnosis sometimes the provocation test is mandatory. Oral and bronchial challenges can be dangerous, while nasal challenge is safer even if must be better standardized. Patients must not use antiinflammatory drugs with the same mechanism of action of ASA; COX-2 inhibitors are generally well tolerated. Antileukotrienes are useful to treat asthma, in association with steroids. Desensitization can be used in very selected patients. PMID- 12908377 TI - Clinical challenges of bleeding in percutaneous coronary intervention. PMID- 12908378 TI - Economic impact of bleeding complications and the role of antithrombotic therapies in percutaneous coronary intervention. AB - The economics of bleeding complications and the role of antithrombotic therapies in percutaneous coronary intervention (PCI) are discussed. More than 1 million PCI procedures are performed annually in the United States, at a mean cost of hospitalization of approximately $9,000 and billions of dollars in total health care costs. Ischemic complications have been reduced to the point that bleeding has become the most common complication. Bleeding complications and transfusions are also among the most costly complications in PCI, accounting for an incremental cost of hospitalization after PCI that may exceed $10,000, due to increased length of stay and the use of additional resources such as ultrasound evaluation and surgical repair of the vascular site. Anemia and transfusions are also associated with increased morbidity and mortality, contributing to additional treatment costs beyond those directly attributable to correcting the bleeding complication. In the past decade, significant reductions in heparin dose and warfarin use were associated with reduced bleeding complications, but glycoprotein IIb/IIIa inhibitors have been shown to increase the clinical and economic costs of bleeding complications. The replacement of heparin with bivalirudin is associated with significant reductions in the costs of antithrombotic therapy and in complications. Reductions in bleeding complications have become a primary target for further improvements in both clinical and economic outcomes. PMID- 12908379 TI - [The quality of medication orders--can it be improved?]. AB - Medication errors are a common cause of morbidity and mortality among patients. Medication administration in hospitals is a complicated procedure with the possibility of error at each step. Errors are most commonly found at the prescription and transcription stages, although it is known that most errors can easily be avoided through strict adherence to standardized procedure guidelines. In examination of medication errors reported in the hospital in the year 2000, we found that 38% reported to have resulted from transcription errors. In the year 2001, the hospital initiated a program designed to identify faulty process of orders in an effort to improve the quality and effectiveness of the medication administration process. As part of this program, it was decided to check and evaluate the quality of the written doctor's orders and the transcription of those orders to the nursing cadre, in various hospital units. The study was conducted using a questionnaire which checked compliance to hospital standards with regard to the medication administration process, as applied to 6 units over the course of 8 weeks. Results of the survey showed poor compliance to guidelines on the part of doctors and nurses. Only 18% of doctors' orders in the study and 37% of the nurses' transcriptions were written according to standards. The Emergency Department showed an even lower compliance with only 3% of doctors' orders and 25% of nurses' transcriptions complying to standards. As a result of this study, it was decided to initiate an intensive in-service teaching course to refresh the staff's knowledge of medication administration guidelines. In the future it is recommended that hand-written orders be replaced by computerized orders in an effort to limit the chance of error. PMID- 12908380 TI - [Non-compliance with medication as a common reason for admission to a closed psychiatric ward]. AB - Noncompliance with pharmacologic treatment is common amongst patients suffering from chronic diseases. In psychiatric patients compliance is extremely important in order to prevent recurrence of the disease. Recurrence is associated with readmissions and increased costs to the health system. About 70% of patients suffering from schizophrenia will relapse within a year without medication. This study evaluates the causes of admission and diagnoses of 100 patients consecutively admitted to a closed psychiatric ward between 1.4.2000 and 23.6.2000. Fifty- four of these 100 admissions were associated with medication noncompliance in the last year. Noncompliance was not associated with psychiatric diagnosis, gender, type of medication or form of administration of the medication. The problem of noncompliance should be approached on two levels: the doctor-patient relationship and reorganization of mental health services. On the doctor-patient level, the therapist has to maintain a therapeutic alliance with the patient and his family and adjust the medication accordingly, in order to maximize the benefit and minimize side effects. On the health system level, mental health should be included in the National Health Insurance Law and the Health Funds should take responsibility for mental health as in other medical fields. This may enable the psychiatric patients to be under the care of their general practitioner in continuity with the psychiatric hospital. Thereby, resources should be transferred from the psychiatric hospitals to the community. The availability of the psychiatric community services and their links to the general healthcare system would increase compliance and reduce recurrence, readmissions and the stigma of patients suffering from psychiatric disorders. PMID- 12908381 TI - [Diode laser cyclophotocoagulation in refractory glaucoma]. AB - PURPOSE: To evaluate the effect of contact transscleral diode cyclophotocoagulation for refractory glaucoma. METHODS: Thirty-three patients with intractable glaucoma were treated with contact diode cyclophotocoagulation. Laser radiation (810-nm) was delivered to the eye by a fiberoptic (G-probe) placed 1.2 mm behind the limbus. Laser setting used was 2 W power for the duration of 2 seconds. Forty to 45 laser spots were applied over 360 degrees. Intraocular pressure (IOP), the number of glaucoma medications, visual acuity and complications were all assessed at last follow-up visit 12 months after laser treatment. Success was defined as IOP of less than 22 mmHg with medications at the last follow-up visit. RESULTS: The mean IOP decreased from 37.8 +/- 8.0 mmHg pre-diode treatment to 18.2 +/- 9.2 mmHg at last follow-up visit after treatment (p < 0.05). About 73% of patients achieved IOP of between 2 and 22 mmHg at final follow-up. The mean number of medications required decreased from 3.4 +/- 1.1 before to 2.4 +/- 1 after treatment (p < 0.05). Fifty five percent of the patients had a deterioration in visual acuity. Other complications were few and included transient hyphema, uveitis, choroidal detachment and hypotony. CONCLUSION: Contact transscleral diode cyclophotocoagulation is effective in lowering IOP and reducing the number of glaucoma medications in refractory glaucoma. The deterioration in visual acuity post-laser is a limitation of the treatment. PMID- 12908382 TI - [Autoimmune hepatitis during pregnancy]. AB - BACKGROUND: Autoimmune hepatitis is a rare chronic disease that mainly affects young women and may influence fertility and pregnancy in these patients. OBJECTIVES: To describe pregnancy and labor in a patient suffering from autoimmune hepatitis and to review the relevant literature. METHODS: Computerized literature research. RESULTS: The disease of 9 years' duration did not relapse during pregnancy under continuous treatment with steroids, cytotoxic drugs, ursodeoxycholic acid and vitamins. Pregnancy was uneventful and ended with preterm vaginal delivery at 35 weeks, of a 2299 gram healthy neonate. The patient did well during the postpartum period and 6 months thereafter. Since there is no single diagnostic test for the disease, the diagnosis is based on the combination of clinical, laboratory, and histopathological findings and by exclusion of other causes of hepatitis. CONCLUSIONS: There is paucity of data in the literature on pregnancy in these patients but the treatment protocols seem to be effective for the mother and safe for the fetus. Maternal and fetal complications rates appear to be currently lower than in the past. Given that relapse cannot be predicted, close surveillance during pregnancy is warranted. PMID- 12908383 TI - [Anticoagulation in pregnant women with prosthetic heart valve--a new approach for therapy]. AB - The treatment of women in childbearing age with a mechanical heart valve is a challenge for the medical staff. Warfarin (Coumadin) is considered to be a safe and effective anticoagulant for patients with prosthetic heart valves. However, treatment during pregnancy poses many difficulties, especially during the first trimester, due to its ability to cross the placenta and its associated fetotoxicity. Treatment with heparin during the first trimester decreases the rate of embryopathy, but increases maternal morbidity and mortality. Warfarin therapy throughout pregnancy, which is common mainly in Europe, carries low rates of maternal complications and roughly six percent of embryopathy. Several studies compared warfarin treatment throughout pregnancy versus treatment with heparin during the first trimester. The relationship between daily warfarin doses and the rate of embryopathy was recently investigated. We report two cases of pregnant women with mechanical heart valves who were treated with heparin during the first trimester. Both underwent an emergency replacement of the prosthetic valve during the eighth week of pregnancy. In this article, we review the literature regarding anticoagulation therapy in pregnant women with prosthetic heart valves; the comparison between treatment with warfarin throughout pregnancy and heparin in the first trimester; and the relation of daily warfarin doses with the rate of embryopathy. The two case reports demonstrate the common approach for therapy and the danger within it. In the discussion we present a new approach for treating pregnant women with prosthetic valve and guidelines for the medical staff. PMID- 12908385 TI - [Diffuse lichen sclerosus et atrophicus involving the palms]. AB - Lichen sclerosus et atrophicus (LSA) is an extremely rare condition which usually involves the genitalia. There are sporadic reports of diffuse LSA with hand involvement apparently the rarest. Although the clinical course of the disease is often benign, there may be widespread lesions, disabling joint contractures, and cutaneous malignancies. We report on a patient in whom diffuse LSA appeared in the sixth decade. The condition led to multiple aggressive squamous cell carcinoma in the upper limbs and other anatomic sites and severe deforming and disabling contractures of the fingers and palms which prevented the patient's use of his hands even for simple tasks. On the basis of our experience, we suggest the application of aggressive primary treatment for cutaneous lesions in LSA. PMID- 12908384 TI - [Cyclic psychosis associated with the menstrual cycle]. AB - Cyclic psychosis associated with the menstrual cycle is an uncommon disorder, not included under the accepted definitions of functional psychoses. We present three female adolescents who developed an acute psychosis a few days before menstruation, which resolved completely upon bleeding or several days later, only to reappear in the same form in subsequent cycles. The clinical presentation was not in line with that of the typical functional psychoses. An extensive medical work-up did not show any significant disturbances, with the exception of anovulatory cycles in one youngster. Psychotropic treatment had no effect on the course of the psychosis. Treatment with progesterone in the second half of the cycle in one case, and with a combined progesterone/estrogen contraceptive agent in another, resulted in full recovery within several cycles. The third girl showed a spontaneous remission within four cycles. Remission continued in all cases after discontinuation of the hormonal treatment, and with no need to reintroduce any psychotropic agent, for a period of 2-4 years. We discuss several possible etiologic mechanisms for cyclic psychosis associated with the menstrual cycle, including it being a cycloid non-specific affective disorder, and its association with a temporary functional hypothalamic-pituitary dysfunction, and with anovulation. We also discuss the role of psychotropic and hormonal treatment in this disorder. PMID- 12908387 TI - [Computerized physician order entry--an urgently required modality]. AB - Misread or misinterpreted hand-written prescriptions or medication orders are the most important cause of life-threatening or serious drug related adverse effects occurring in hospital in-patients. There is convincing evidence in the literature showing that computerized systems for physician order entry, with on-line screening of drug related orders for compatibility with patient characteristics (e.g. age, renal function), drug information (e.g. dose limits, drug-drug interactions) and sets of institutional rules of performance can reduce rates of medication related errors by 80% and more. Reduction of error-related hospital costs would generate the funding required for implementation of such systems, a need that must be met without delay. PMID- 12908386 TI - [The IDF evacuation from southern Lebanon in 2000: impact on the delivery of health care to displaced Lebanese population]. AB - BACKGROUND: The State of Israel controlled a protective security zone in southern Lebanon from 1982 until May 2000, when the Israel Defense Forces (I.D.F.) evacuated this area within two days. Officers and soldiers of the South Lebanese Army, a local militia, which had formerly cooperated with the I.D.F., chose to come to Israel with their families. The provision for the health needs of this displaced population of over 5,861 persons was coordinated by the Northern Region Health Office of the Ministry of Health. AIM: To describe the deployment and provision of health care for this displaced population and to present the lessons learned. METHODS: Data was collected from interviews with the personnel most actively involved in the decisions and execution of the enterprise, as well as from relevant documents. RESULTS: The main health needs of the displaced South Lebanese were related to anxiety, outbreak of chickenpox, dental problems and provision of medical equipment for the disabled. CONCLUSION: The experience accrued by the Northern Region Health Office over the years in provision of care to displaced populations at short notice was implemented. Cooperation within the health sector and between the health services and the military and defense forces was invaluable and essential to deal successfully with this unusual situation. PMID- 12908388 TI - [Confidential healthcare for Israeli adolescents: towards evolution of specific legislation]. AB - BACKGROUND: Despite its critical importance, the issue of confidentiality in the provision of adolescent health care has not been specifically clarified by Israeli law. OBJECTIVE: To review the conflict between the needs of adolescents, attitudes of the medical community and daily medical practice, and to promote legislation that would allow health care providers to treat adolescents without parental consent under certain circumstances. THE CURRENT CONFLICT: A major key to the productive encounter between the adolescent and the health professional is the confidence that sensitive information, communicated between them, will not be divulged to anyone, including parents. The Israeli law does not allow minors to give independent consent to care, except under certain circumstances. Also, Israeli physicians frequently do not provide confidential care, even when the issue is explicitly covered by the law such as in the case of termination of pregnancy. STEPS TOWARDS CHANGE: The Ministry of Health adopted an analogy stating that "in the case of an activity in which minors of his/her age are accustomed to engaging, a legal act performed by a minor cannot be cancelled". On this basis, medical associations and other bodies involved in adolescent health recommended that care related to issues requiring privacy should be provided to minors without parental consent or knowledge whenever it is common among a certain age group to seek the specific type of care. However, a clear and updated legislation is still required in order to enhance professional legitimization and awareness. PMID- 12908389 TI - [Molecular markers for transitional cell carcinoma of bladder]. AB - Superficial transitional cell carcinoma (TCC) has significant rates of recurrence and progression, therefore accurate urinary makers for early diagnosis and follow up are essential. Urine cytology is the oldest marker, constantly used clinically and considered the "gold standard". In this review we will summarize the properties of different molecular markers for TCC and their yield compared to cytology. Briefly, the older markers--BTA stat, BTA TRAK and NMP22 are more sensitive than cytology, but their specificity is lower, especially in patients with underlying urologic pathologies. FDP (fibrinogen degradation products) analysis is rapid and the test is available clinically. It's sensitivity is higher than cytology for low grade tumors, but specificity is low. Of the newer markers, telomerase has a high sensitivity, and a specificity similar to urine cytology, but also has an increased rate of false positive results. HA-HAase (Hyaluronidase) and CK20 have promising sensitivity and specificity, but are still unavailable for clinical use. CK20 has the additional advantage of being able to detect premalignant conditions as well as predicting recurrence. The role of urinary markers in replacing cystoscopy and urine sediment cytology in the follow-up of TCC and in hematuria work up is still uncertain. It is possible that urinary markers in different panel arrays will have a role in the future for those indications and perhaps in the screening of populations at risk, such as smokers and workers exposed to occupational hazards. PMID- 12908391 TI - [NNT--numbers needed to treat]. AB - Number needed to treat--NNT--is a bridging tool between statistical results from clinical trials and the physician daily work. This number represents the number of patients we need to treat with the experimental therapy in order to prevent one of them from developing the bad outcome. The concept is a useful measure of the clinical effort our patient and we must expend in order to help them avoid bad outcomes to their illness. For example, the NNT for antibiotic treatment of otitis media is 8, which means that we have to treat eight children with antibiotic and only one out of the eight will benefit the treatment. In a medical world that uses the Evidence Based Medicine as essential tool, we believe that the NNT is an important tool and very efficient for choosing the best treatment method. PMID- 12908390 TI - [Silicone breast implants with silicone gel and autoimmune disease--are they related?]. AB - Silicone breast implants are used for breast reconstruction and breast augmentation. In the late 80's anecdotal literature describing a possible connection between silicone gel filled breast implants and autoimmune disease accumulated. Emerging concern about the safety of silicone gel implants had led the FDA to restrict their use. The continued use of the implants in postmastectomy reconstruction was allowed, as was their use in a small number of breast augmentation patients who were willing to enroll in long-term studies. In recent years, large meta-analyses established the fact that there is no association between silicone gel breast implants and autoimmune disease. We review the current literature, emphasizing the most important epidemiologic studies. PMID- 12908392 TI - [Binge eating disorder (BED)--new diagnostic category]. AB - Binge Eating Disorder (BED) is a proposed new diagnostic criteria in the DSM-IV, cited as an example of an eating disorder Not Otherwise Specified (NOS). The phrase Binge Eating was introduced over 35 years ago to describe problematic behavior found in a subgroup of overweight individuals, and as a subgroup of Bulimia Nervosa. The defining characteristics of BED patients are consumptions of an objectively large amount of food in discrete time periods, a lack of control during eating episodes and a lack of compensatory behavior binge. BEDs affect approximately 30% of obese individuals seeking treatment. The binge eaters represent a distinct subgroup among the obese with higher prevalence of comorbid psychiatric disturbances, as well as higher rates of eating pathology, as compared to normal obese women. Several researches suggested that individuals use binge eating as a means of coping with emotional distress. In addition, the binge eating disorder represents a distinct subgroup among bulimia nervosa with lower comorbid psychiatric disturbances, eating pathology and purging behavior. The present paper will review this proposed syndrome and will discuss the question of whether it is a distinct subgroup of obese patients or a distinct subgroup of bulimia nervosa patients, or a new diagnostic category. This diagnostic dilemma is discussed in light of clinical and empirical literature. Clinical and therapeutic implications are presented and ideas for future research are proposed. PMID- 12908393 TI - [Salutogenesis--a story about the development of an idea]. AB - The medical sociological concept of salutogenesis is introduced and explained. Salutogenesis means essentially: what factors create and maintain relative health; this is in contradiction to the well known questions on pathogenesis. There is special emphasis on the central component: the sense of coherence and the three sub-categories--comprehensibility, manageability and meaningfulness. The importance of the concept salutogenesis for the theory of general medicine and psychotherapy is explored. An eventual application of this concept and attitude in practical medicine is suggested and discussed. The possible importance of the salutogenesis concept and attitude, for coping with continuing and intense professional stress of physicians and other therapists, is outlined. PMID- 12908394 TI - [The identification of the immunodominant gliadin epitope]. AB - Recent efforts have piri-pointed a relevant sequence on gliadin, that may act as the immunodominant epitope in celiac disease. Independent reports from several leading laboratories identify a 33 amino acid segment spanning through residues 57-89, and excessively rich in proline and glutamine, as the putative epitope. A glutamine residue in position 65 is the site of deamidation by transglutaminase, promoting its recognition by DQ2-HLA. In addition, another gluten protein, glutenin, may be relevant to the pathogenesis in celiac disease. PMID- 12908395 TI - [The terminal patient: Jewish religious law, the Steinberg report and the bioethical discourse in Israel]. AB - This article surveys key texts in contemporary orthodox Jewish law (Halakha) with regard to end-of-life decision making. The author proposes twelve principles that govern Jewish law in that matter. The article proceeds to examine the Steinberg report in the light of Halakha. Orthodox Judaism regards human life as a prime value, which is always beyond consideration of economical means or quality of life. The avoidance of suffering is the only justification to shorten the life of the sufferer, provided that the acts performed do not fall within the Halakhic definition of murder, namely active and direct action that shortens life. It is argued that the main challenge of bioethics in Israel is the bridging between the positive law of Halakha whose fundamental value is submission to God's will as manifested in Halakha, and the rationalism, universalism, and egalitarianism which constitute naturalistic ethics. This challenge may produce ideas such as the "clock machine". It is too early to know if this is a trickery, or genuine ethical creativity. PMID- 12908396 TI - [Characteristic of combat trauma and treatment of gunshot fractures of long bones of the limbs]. AB - Treatment of the patients with gunshot fractures of extremity long bones is one of the main problems of field surgery. The complex study of medical records obtained from 718 servicemen with gunshot fractures of extremity long bones who participated in counter-terrorist operation in the Republic of Chechnia (1994 1996) allowed to evaluate character and severity of the injuries, content of the treatment measures conducted at the stages of medical evacuation. The peculiarities of fighting trauma were the following: the high frequency of combined (22.5%) and multiple (25.9%) injuries, the high share of multi fragmentation and splintered (76.4%), intra-articular fractures (17.3%), the primary defects of bones (7.1%) and soft tissues (4.8%), the injuries of main vessels (12.1%) and nerves (18.5%). Owing to the short periods of evacuation to the stage of specialized medical care (up to 18.2 +/- 5.3 h), high share of preserving variant of primary surgical treatment (82.3%); high quality of medical immobilization at the expense of wide introduction of functionally stable external osteosynthesis (51.2% of the casualties) it was possible to improve the treatment results. PMID- 12908397 TI - [The information security and its urgency for military medical service]. PMID- 12908398 TI - [Surgical treatment and rehabilitation of the patients with post-traumatic nose and upper lip defects and deformities]. PMID- 12908399 TI - [Psychological risk factors in development of cardiovascular diseases in young military personnel]. AB - The results of investigation using the routine examinations and psychological methods have revealed the psychological risk factors of development of cardiovascular diseases in the healthy young servicemen. It was established that the disorders in formation of psychological adaptation mechanisms (P < or = 0.05) should be considered as such risk factors. To decrease the probability of cardiovascular pathology in servicement it is necessary to use the methods of psychological correction and psychotherapy. PMID- 12908400 TI - [Changes in peroxide homeostasis in servicemen during the adaptation to the military service and climatic-and-geographical conditions]. PMID- 12908401 TI - [Pseudotumorous form of yersiniosis]. PMID- 12908402 TI - [Iodocasein in the prophylaxis of iodine deficiency in servicemen]. AB - To evaluate the efficiency of iodcasein in prophylaxis of iodine-deficient states in servicemen the following parameters were studied: iodine sufficiency determined by ioduria median; thyroid volume measured on the base of US data; functional state of hypothalamo-pituitary-thyroid system evaluated by blood level of thyroid hormone and free thyroxin; physical performance assessed by veloergometric test, auto-and-verbal memory and attention change-over. Iodine deficiency was initially detected in 88.9% (main group) and in 94.4% (control group). Ioduria median corresponded to iodine deficit of middle severity in both groups. After the initial examination the persons from the main group were given iodized bread during 9 months. The monitoring results have confirmed the efficiency of iodine prophylaxis with the use iodcasein. PMID- 12908403 TI - [Prophylactic measures in the restoration of occupational health of air force personnel]. PMID- 12908404 TI - [Survival and recovery from alimentary dystrophy and psycho-emotional stress in Leningrad blockade]. PMID- 12908405 TI - [Training arrangement for military doctor's assistants]. PMID- 12908406 TI - [20 years of The Central Navy Clinical Hospital]. PMID- 12908407 TI - [The half-century anniversary of the Seliatin military hospital]. PMID- 12908408 TI - [Tarkhovka military health sanatorium]. PMID- 12908409 TI - [Therapeutic effect of some antibiotics on experimental staphylococcal infection and its correlation with in vitro activity of antibiotics in sub-inhibitory concentration against Staphylococcus aureus strains]. AB - The aim of the study was to demonstrate of whether the therapeutic effects of antibiotics depend on their in vitro activity in sub-inhibitory concentrations against staphylococci. Cloxacillin, gentamicin and lincomycin were used in the study. Groups of S. aureus strains, containing 6 strains with similar MIC values each but different sensitivity to sub-inhibitory antibiotic concentrations (sub MIC) were selected (a total of 36 trains): i. strains increasing their sensitivity to phagocytosis and bactericidal activity of rabbit leukocytes after incubation with an antibiotic in 0.1 MIC concentration, ii. strains with sensitivity to the above factors unaffected by incubation with an antibiotic in 0.5 MIC concentration. The doses of staphylococci causing death of 90-100% of Swiss albino mice 10 days after i.p. infection were determined. The injected doses (LD 90-100) and various doses of antibiotics were used to determine ED50 values as well as the survival rate of the mice with experimental staphylococcal infections after treatment with these antibiotics. It was demonstrated that effective doses (ED 50) of the antiboitics were significantly lower when the antibiotics were administered once to mice infected with strains S. aureus sensitive to sub-MIC concentrations of the investigated antibiotics than for mice infected with strains resistant to their sub-MIC concentrations. Similar correlations were observed in mice which were given the antibiotics several times (for 7 days): the percentage of the surviving mice was higher in the group infected with sub-MIC sensitive strains. The therapeutic effect of cloxacillin, gentamicin and lincomycin demonstrated a significant correlation with the S. aureus strains used to induce the infections and their sensitivity, or lack of sensitivity in vitro, to phagocytosis and bactericdal activity of leukocytes in the presence of antibiotics in sub-MIC concentrations. PMID- 12908410 TI - [Infections with Van B phenotype Enterococcus faecium and E. faecalis in patients with immune deficiency during the course of hematologic neoplasms]. AB - In this work, we have analyzed the second outbreak of VRE with the VanB phenotype to be identified in the country. The aim of this study was to characteristics of the types of resistance to glycopeptide antibiotic and to check the resistance patterns of these pathogens. A trial of monitoring the risk factors for colonization or infection with VRE as well as epidemiological investigation were undertaken. Genus identification of the isolates was performed according to the method of Facklam and Collins, and species were identified using the API 20 Strep test. MICs of different antimicrobial agents were determined by the E-test method. The isolates collected during the investigation demonstrated resistance to multiple antimicrobials, which is a common characteristic of VRE. Isolates were found to be uniformly resistant to penicillin, fluoroquinolones, tetracycline and to high concentrations of aminoglycosides. The only drugs with in vitro activity against the isolates were ampicillin (VRES), linezolid (VRES, VREM) and quinupristin-dalfopristin (VREM). Except for a single VREM isolate, they all revealed the typical VanB phenotype with resistance to vancomycin and susceptibility to teicoplanin. One of the VREM isolates turned out to be resistant to teicoplanin, which coincided with the use of this antibiotic in the patient's therapy. Its vanB gene variant differed by a single mutation from that found in other isolates; however, it also lacked a large part of the vanB gene claster, including the regulatory genes vanRB and -SB, and the vancomycin- inducible promoter PYB. Our studies have found an association between colonization or infection with VRE and the mean duration of hospital stay, previously administration of glycopeptide, cephalosporins and imipenem. These organisms were a common cause of monoethiological bloodstream infections. PMID- 12908411 TI - [Expression of proteins regulated by iron levels in enterococci]. AB - Enterococci respond to iron deprivation in vitro by increasing the expression of a number of iron-regulated proteins. They detected in whole protoplasts lysates and corresponded to proteins with apparent molecular masses of the region 14.4-43 Kda. Their occurrence were correlated to siderophore production. PMID- 12908412 TI - [Correlation between the presence of aggregative fibria and a type of Escherichia coli adhesion]. AB - Enteroaggregative strains of E. coli (EAEC) are an important agents possessing among many virulence factors, aggregative fimbria AAF hemagglutinating in the presence of mannose human group A or/and rats erythrocytes. The aim of the study was to determine the correlation between the presence of AAF fimbria and pattern of adherence in vitro. Tested strains of E. coli were obtained from children with diarrhea (133 strains) and healthy children (105 strains). Among strains of E. coli from children with diarrhea 81 (61%) showed the presence of AAF fimbria and 19 (23%) were adhering in aggregative pattern. In the group of strains of E. coli isolated from healthy children 31 (30%) were AAF positive and 8 (25.8%) of them presented aggregative adherence. Examination of AAF fimbria only dose not allow to distinguish EAEC strains. The data showed the participation of EAEC strains in diarrhea of children below 3 years old. PMID- 12908413 TI - [Survival of Bacillus anthracis spores in baths using modern technologic tannery processes]. AB - The influence of skin tannery baths, according as wet-blue, wet-white and plant technology, on B. anthracis spores survival was investigated. As a result of this study there was explained that lime bath do not inactivated of all spores that are present in infected bath. As a result of these experiments the spores were inactivated completely not before pickle bath. PMID- 12908414 TI - [Experimental infections and colonization of porcine skin samples with B. anthracis]. AB - In this study there was demonstrated the process of appearance and intensity of colonisation of the swine's skin samples by exposing them on agar plate cultures or immersed in liquid cultures of B. anthracis. The results of research were documented with photos of coloured histological preparations. As a result of the research there was demonstrated the possibility of infection and colonisation of skin consignment by B. anthracis from accidental located skin of died or killed animals because of anthrax. This colonisation may occur in the period 3 to 24 hours in optimal conditions. Anthrax bacteria are starting to penetrate and colonize the skin samples during 3 hours with exposing them from the hypodermic side; whereas penetration and colonisation from epidermis side appears after 24 hours. The results demonstrated that in natural conditions for optimal cultivation of B. anthracis there can be a possibility of infection and colonization of health parts of the skin's in storage skin for the longer period of time or longer transport. PMID- 12908415 TI - [Comparison of AP-PCR methods, ribotyping (PCR-ribotyping) and pulsed-field electrophoresis (PFGE) for strains of Clostridium difficile producing toxin B and not producing toxin A]. AB - In this study were used AP-PCR, PCR-ribotyping and pulsed-field elecrophoresis (PFGE) for comparative study of toxin A-negative/toxin B-posi-tive Clostridium difficile strains with deletion in toxin A gen. We investigated nine unrelated clinical strains, isolated from different units and different time from patients suffering to antibiotic associated diarrhea (AAD). We found that toxin A negative/toxin B-positive C. difficile strains isolated in Poland belonging to a single genotype A, are being similar to the Japanese strains. PMID- 12908416 TI - [Detection of ermB gene responsible for high level resistance to clindamycin (MLS type resistance) among Clostridium difficile strains isolated from antibiotic associated diarrhea (AAD)]. AB - In 68 C. difficile strains isolated from feacal samples of patients with antibiotic associated diarrhoea (AAD) investigated presence of ermB gene transferable of high level resistance to clindamycin. The primers set 2980/2981 used for identification of ermB gene amplified a 688 bp segment. We used the Etest to assess all strains for susceptibility to clindamycin. This study demonstrates that 57% of strains isolated from faecal samples of patients with AAD were highly resistant to clindamycin (minimal inhibitory concentration (MIC) of clindamycin, 256 mg/L) and possessed the ermB gene. PMID- 12908418 TI - [Susceptibility to antibiotics of bacteria from genera Micrococcus, Kocuria, Nesterenkonia, Kytococcus and Dermacoccus]. AB - Two hundred and nineteen strains from genera Micrococcus, Kocuria, Nesterenkonia, Kytococcus and Dermacoccus isolated from different sources, such as saliva, skin of palm and forearm and from vestibule of nose were tested. The susceptibility to doxycycline, cetriaxone, cefuroxyme, amikacyne, amoksycillin with clavulanic acid, trimethoprim/sulfamethoxazole, ampicillin, ciprofloxacin, penicillin and erythromycin were estimated. In general, bacteria from these genera are sensitive to most of selected antibiotics. Most of the strains showed resistance to ampicillin and erythromycin. None of these strains produced beta-lactamases. In infectious caused by bacteria from genera Micrococcus, Kocuria, Nesterenkonia, Kytococcus and Dermacoccus in clinical treatment should be used amoksycillin with clavulanic acid, doxycycline, cetriaxone, cefuroxyme, or amikacine because, with one's own range of activity embrace highest percentage of investigated strains. PMID- 12908417 TI - [Occurrence and number of bacteria from the Micrococcus, Kocuria, Nesterenkonia, Kytococcus and Dermacoccus genera on skin and mucous membranes in humans]. AB - The aim of work was to evaluate the frequency of occurrence of bacteria from genera of Micrococcus, Kocuria, Nesterenkonia, kytococcus and Dermatococcus on human skin and mucous membranes in healthy population. Among 150 investigated persons these bacteria were found in 122 (81.3%). The frequency of isolation was similar in both sex (82.4% in female and 79.7% in male). Most often the strains were isolated from oral cavity (48.7%), and from skin of palm and forearm (40.7% and 37.3%). Least frequency of occurrence was observed in vestibule of nose (26%). The predominant isolated strains were: M. luteus (26.2%), and N. halobia (21%) followed by K. varians (16.4%), M. lylae (12.2%), D. sedentarius (9.1%), K. kristinae (7.3%), K. nishinomiyaensis (7.3%), K. rosea (0.3%). PMID- 12908419 TI - [Interleukin 6 in neutrophil cultures and mononuclear cells in blood serum of patients with Lyme disease]. AB - The aim of this study was to estimate the release of IL-6 by human neutrophils (PMN) and peripheral blood mononuclear cells (PBMC) in patients with Lyme disease confronted with the serum levels. The cells were isolated from whole blood of 25 patients and of 10 healthy donors and cultured in the presence of LPS. In the culture supernatants and serum the concentration of IL-6 with ELISA (BioSource) were measured. In patients we observed higher values of IL-6 released by unstimulated PMN and PBMC in compared with control. In contrast to control, we didn't observe increased the release of IL-6 by LPS-stimulated PMN and PBMC as compared to unstimulated cells. In the serum of patient we found increased the concentration of IL-6. The higher ability of PMN and PBMC from patients with Lyme disease to release of IL-6 and the lack response to additional stimulation indicate the activation of PMN and PBMC in vivo. PMID- 12908420 TI - [Use of impedance test for testing effectiveness of drug preservatives]. AB - Pharmacopoeias contain preservation efficacy test for estimating antimicrobial activity of chemical compounds added to pharmaceutical preparations in multidose containers in order to inhibit bioburden growth. This method involves the treating of preserved products with bacteria and yeast cells and monitoring the survival of microorganisms through the specified time periods up to 28 days. The last stage of assay--incubation and colony counting--is very time consuming. Recent advance in technology enables faster and more convenient detection in comparison to traditional methods. Impedance method is based on the principle that conductance and capacitance of cultivation medium increases when bacteria grow and metabolize, Impedance time detection is inversely proportional to initial bacterial population. Six different products were utilised throughout the study. The calibration curves were calculated for each of the tested strains by comparison between standard plate count method and detection time measured in Bactometer system. In our study log reduction calculated in alternative method were similar to those obtained in plate count assay. All of the tested preparations, except one, exhibited acceptable activity against bacteria and fungi and meet the pharmacopoeal requirements. The studies indicated a positive correlation between standard plate count results and impedance reading. The procedure with the usage of Bactometer, provides a rapid and accurate system for the determination of bacterial content. PMID- 12908421 TI - [Use of impedance methods for testing bactericidal activity of antiseptics]. AB - Standards PN-EN 1040 and EN 12054 describe test methods and minimum requirements for bactericidal activities of antiseptics. However, standard procedures are time consuming and require 48 hours of incubation. New alternative technique based on impedimetric procedure provides possibilities to reduce this time to several hours. The Bactometer (BioMerieux, Vitek System, USA) is a fully automated impedance technology system used to microbial quantitation of products. Impedance measures microbial activity by electrical methods. The aim of the study was to adaptate the impedimetric method utilising Bactometer--system to microbiological activity control of chemical antiseptics. Eight different products were utilised throughout the study. The samples for classical and alternative method were prepared in the same way as described in standards. The method of choice was dilution-neutralization method. All procedures conducted in Bactometer were verified by plate count method. The high correlation was observed between results obtained by normative methods and impedimetric measurement. All tested products meet requirements. The procedure utilising the Bactometer, provides a rapid and accurate system for the determination of bacterial content. The results of validation carried out during this study indicate, possibility to use impedimetric method alternatively to traditional methods. PMID- 12908422 TI - Nanoporous materials. AB - The current review article attempts to cover the field of nanoporous materials, thereby materials with voids of the order of 1 nm to 100 nm or even larger. Concepts, that have an effect on a more general basis, are introduced and the common features among the different classes of nanoporous materials are demonstrated enabling understanding their formation and stability. It is seen that the most successful method to produce nanoporous materials made by design is to imprint a template structure into a functional inorganic or even organic network. Further, the manuscript gives an introduction to state of the art overview for microporous, mesoporous and macroporous materials, while the preference lies on ordered pore structures. Further, it is demonstrated how important but also demanding the use of modern analytical techniques are. This makes the field of nanoporous materials truly interdisciplinary with aspects of inorganic chemistry, polymer science, colloid chemistry, physical chemistry, analytical chemistry and chemical engineering. PMID- 12908424 TI - Novel synthesis of polymorphous nanocrystalline KNbO3 by a low temperature solution method. AB - Orthorhombic and rhombohedral single nanocrystalline KNbO3 with different morphologies was synthesized respectively at a relatively low temperature by simply changing the reaction solvent; a conceivable reaction process was illustrated based on the finding of a hexahedral intermediate product during the formation of KNbO3. PMID- 12908423 TI - DNA-directed magnetic network formations with ferromagnetic nanoparticles. AB - We formed a DNA network embedding ferromagnetic cobalt nanoparticles with a 12-nm diameter through a nanoscale self-assembly of DNA molecules on large-scale mica surfaces (12 mm x 12 mm); we then confirmed its structural characteristics with an atomic force microscope. Moreover, noncontact magnetic force microscope measurement revealed that some embedded cobalt nanoparticles have different directions of magnetization, similar to "bits" in magnetic data storage devices. PMID- 12908426 TI - Fabrication of ordered arrays of InP microstructures by wet chemical etching with Au masks. AB - Ordered arrays of InP microstructures have been fabricated on InP(001) substrates by wet chemical etching in aqueous HCl with patterned Au masks. The masks were produced by Au deposition through copper grids or a monolayer of polystyrene microspheres. Square InP mesas (20 x 20 microns) and pillars (approximately 100 nm in both diameter and height) were both produced and characterized by scanning electron microscopy and atomic force microscopy. PMID- 12908425 TI - Raman-active modes of single-walled carbon nanotubes derived from the gas-phase decomposition of CO (HiPco process). AB - Here we report Raman scattering studies of ropes of Single-walled carbon nanotubes (SWNTs) grown by a high CO pressure process. Five samples from five different batches were studied as a function of excitation wavelength. Three of these samples exhibited Raman spectra similar to that found for SWNTs made by pulsed laser vaporization of arc-discharge methods. The other two samples were found by Raman scattering to contain a significant fraction of tubes with diameter < 1.0 nm. These samples exhibited unusual spectra that, however, can be well understood within the existing models for the electronic and phononic states in SWNTs. Spectra recorded with 1064 nm for the sample having a significant fraction of smaller diameter tubes shows strong modes present between 500 and 1200 cm-1. We suggest these modes arise due to the enhancement of Raman cross section for small diameter tubes. PMID- 12908427 TI - A study of polyaniline-carbon nanotube composites. AB - Composites of polyaniline (PANI) have been prepared with pristine multiwalled and single-walled nanotubes as well as nanotubes subjected to acid treatment and subsequent reaction with thionyl chloride. The composites have been characterized by various techniques, including X-ray diffraction, electron microscopy, as well as infrared and Raman spectroscopy. Electrical resistivities of the PANI-nanotube composites have been measured and compared with those of the nanotubes and PANI. PMID- 12908428 TI - Nanofriction behavior of cluster-assembled carbon films. AB - We have characterized the frictional properties of nanostructured carbon films grown by supersonic cluster beam deposition via an atomic force-friction force microscope (AFM-FFM). The experimental data are discussed on the basis of a modified Amonton's law for friction, stating a linear dependence of friction on load plus an adhesive offset accounting for a finite friction force in the limit of null total applied load. Molecular dynamics simulations of the interaction of the AFM tip with the nanostructured carbon confirm the validity of the friction model used for this system. Experimental results show that the friction coefficient is not influenced by the nanostructure of the films nor by the relative humidity. On the other hand the adhesion coefficient depends on these parameters. PMID- 12908429 TI - Size-dependent absorption and defect states in CdSe nanocrystals in various multilayer structures. AB - GeS2-CdSe superlattices and composite films are prepared by consecutive thermal evaporation of CdSe and GeS2 in vacuum. CdSe layer thickness varies between 1 and 10 nm, while the thickness of GeS2 layers is either equal (in superlattices) to or 20 times greater (in composite films) than that of CdSe layers. Standard spectral photocurrent measurements and various constant photocurrent methods are used to study optical absorption of all samples. An overall blueshift is observed with decreasing CdSe layer thickness of superlattices. This shift is related to a size-induced increase of the optical band gap of CdSe due to one-dimensional carrier confinement in the continuous nanocrystalline CdSe layers. A number of features are observed in the absorption spectra of composite films containing CdSe nanocrystals with average radii of approximately 2.5 and approximately 3.3 nm. They are discussed in terms of three-dimensional carrier confinement and are considered a manifestation of excited electron states in CdSe nanocrystals embedded in GeS2 thin film matrix. In addition to these discrete features, the exponential dependence of the optical absorption (Urbach) edge indicates a distribution of "valence band" tail states associated with disorder. Transient photoconductivity measurements made on similarly prepared SiOx-CdSe superlattices exhibit a rapid fall in photocurrent by a power law decay over several orders of magnitude of time, which is consistent with multi-pletrapping transport via an extensive distribution of deep defects. PMID- 12908430 TI - Nanosensor for in vivo measurement of the carcinogen benzo[a]pyrene in a single cell. AB - This work describes the fabrication and the application of an antibody-based fiber-optic nanosensor for in situ measurements of the carcinogen benzo[a]pyrene (BaP) in a single cell. This antibody-based spectroscopic nanosensor is miniaturized enabling the detection of fluorescent analytes in single cells. In addition to measuring fluorescent analytes in single cells, the nanosensor has the potential to be applied for both diagnostic and proteomics purposes. In this work, the human breast carcinoma cell line, MCF-7, was used as the model system to perform BaP measurements in single cells. A standard concentration curve for BaP was established and used to perform quantitative analyses of BaP in individual cells. From these analyses, it was estimated that the concentration of BaP in the individual cells investigated was approximately 3.61 x 10(-10) M. The results obtained demonstrate the application of antibody-based nanosensors for performing in situ measurements inside a single cell. PMID- 12908431 TI - The effect of substituents on molecular electronic junctions. AB - Previous reports have demonstrated that molecular electronic junctions exhibiting negative differential resistance show a marked substituent effect. We show that this substituent effect correlates with stability of the anionic form of the junction molecule. A sufficiently stable anion gives rise to a double potential barrier to electron transport across the molecular junction. The effect of thermal motion of the junction molecule on the electron transport is also considered. PMID- 12908432 TI - Stoichiometric complexes between cyclic phenylazomethines and a dialkyl phosphate for molecular tiling at the air-water interface. AB - The molecular tiling method is proposed as a novel methodology to provide an artificial molecular arrangement at the air-water interface by assembling rigid amphiphilic units like tile pieces. In this study, the mixed monolayers of cyclic phenylazomethine (CPA) as a rigid triangle basement and dihexadecyl phosphate (DHP) as a monolayer-forming component were investigated by pi-A isotherm measurement and Fourier transform infrared (FTIR) spectroscopy in the reflection absorption mode in order to provide basic knowledge of molecular tiling design. Although the CPA molecules alone cannot form a stable monolayer on water, monolayers of DHP/CPA mixtures showed two-step condensed phases that reflect two kinds of assembling modes of alkyl chains of these components. FTIR spectra of the transferred monolayers indicated specific interaction of the amine part in CPA and the phosphate moiety of DHP. pi-A isotherms of the systematic change in the mixing ratio revealed the binding stoichiometry between DHP and the CPA to be 3:1. Interaction of DHP molecules to three basic sites of CPA and matching in cross-sectional area between three DHP molecules and CPA core would result in the stable formation of the 3:1 complex. PMID- 12908433 TI - Fabrication and properties of spray-dried nanofeatured spherical Ni(OH)2 materials. AB - Spherical agglomerates of nanostructured beta-phase Ni(OH)2 with the general formula Ni1-xCox(OH)2 (x = 0, 0.1, 0.3) for use as cathode materials were produced by a modified method including coprecipitation of Ni or Ni composite hydroxide and further spray drying of the precipitated and washed slurry. This process leads to the formation of spherical agglomerate particles with a narrow Gaussian-type distribution range. The method permits faster and cheaper production of cathode materials with a higher specific surface area and similar or better capacity and cycle life compared with the materials prepared via conventional technology. PMID- 12908434 TI - Computational engineering of metallic nanostructures and nanomachines. AB - Small structures with dimensions in the nanometer regime play an important role within a lot of modern technological branches like, for example, genetics, chip fabrication, material science, medicine, or chemistry. While highly sophisticated characterization methods would be necessary to study such nanostructures, computational methods and models have made their entrance into the field of nanotechnology. The present work gives an overview of the problems connected with quantum mechanics, many-particle systems, and nanophysical models. Further, the application of molecular dynamics (MD)--a typical computational method suitable for modelling at the nanolevel--is introduced and outlined. The setup and use of specific MD models, advanced computation techniques, and efficient algorithms are discussed, while the focus is laid on the subjects nanodesign and nanoengineering which are demonstrated for the example of metallic nanostructures. Finally, the introduced techniques and methods are applied to stability studies of theoretical nanomachines. PMID- 12908435 TI - Molecular dynamics study of hypothetical silicon nanotubes using the Tersoff potential. AB - We have performed classical molecular dynamics simulations for hypothetical silicon nanotubes using the Tersoff potential. Our investigation presented a systematic study about the thermal behavior of hypothetical silicon nanotubes and showed the difficulty in producing silicon nanotubes or graphitelike sheets. However, since the elastic energy per atom to curve the sheet into cylinders for silicon atoms is as low as that for carbon atoms, if graphitelike sheets of silicon are formed, the extra cost to produce the tubes is of a similar order to that in carbon. Through the investigations on the structure and properties of a double-wall silicon nanotube, we concluded that quasi-one-dimensional structures consisting of silicon atoms become nanowires rather than nanotubes in order to minimize the number of sp2 bonds. PMID- 12908436 TI - Heart attack. Fast action needed. PMID- 12908437 TI - Handling happy hour. PMID- 12908439 TI - Get moving with yoga. PMID- 12908438 TI - Making positive changes. PMID- 12908440 TI - Diabetes quiz. How much do you know about dietary fat? PMID- 12908441 TI - Flavoring your food with herbs. PMID- 12908442 TI - Making sense of dietary recommendations. PMID- 12908443 TI - The myth of brittle diabetes. PMID- 12908444 TI - Diabetes definitions. Retinal detachment. PMID- 12908445 TI - Swedish country cooking. PMID- 12908447 TI - Supermarket smarts. Vegetables. PMID- 12908446 TI - Choosing a blood glucose meter. PMID- 12908448 TI - Mechanisms implicated in the growth regulatory effects of vitamin D compounds in breast cancer cells. AB - The active metabolite of vitamin D3, 1,25-dihydroxyvitamin D3 (1,25(OH)D3), has been recognized for over 2 decades as a modulator of cell proliferation and differentiation in many cell types, including breast cancer. However, any potential anti-tumour properties displayed by 1,25(OH)D3 are limited by the tendency to cause hypercalcaemia when administered at high doses. Because of this, synthetic vitamin D analogues have been developed that retain the anti tumour effects seen with 1,25(OH)D3 but which have reduced calcaemic activity. However, it is still unclear as to how 1,25(OH)D3 and its synthetic analogues act within breast cancer cells to elicit the effects on cellular proliferation and differentiation. In this chapter we review the advances that have been made in trying to answer this question. It has been found so far that 1,25(OH)D3 has an effect on the expression of certain cell cycle regulators and in this way can bring about G1 arrest. Evidence has also emerged that vitamin D compounds can also affect the growth-promoting pathways initiated by two important factors involved in breast cancer cell promotion; namely the insulin-like growth factor I (IGF-I) and oestrogen-receptor (ER) pathways. Vitamin D compounds have also been implicated in promotion of apoptosis in breast cancer cells and evidence suggests that 1,25(OH)D3 and its synthetic analogues may potentiate responsiveness of breast cancer cells to conventional cytotoxic agents. Although much remains to be learned about the associated underlying mechanisms, ongoing research suggests that vitamin D analogues are a new class of compounds with potential in breast cancer treatment and prevention. PMID- 12908449 TI - [The Puerto Rico Heart Health Program. 1961-1970]. PMID- 12908450 TI - Applications of RNA1 for Drug Discovery-IBC Conference. 8-9 May 2003, London, UK. PMID- 12908451 TI - Osteoporosis & Menopausal Disorders-SMi Conference. PMID- 12908452 TI - Depression & Anxiety-SMi Conference. Shining light into dark corners. 12-13 May 2003, London, UK. PMID- 12908454 TI - [The use of laser in dermatology. Fascination, advances and catches of a pioneering technology]. PMID- 12908453 TI - [Topical immunomodulators in dermatology]. AB - Immunomodulators include both immunostimulatory and immunosuppressive agents. Obligate contact sensitizers such as diphencyprone or dinitrochlorobenzene have been used against viral and autoimmune diseases. Newer agents such as the toll like receptor agonists imiquimod and resiquimod have been clinically used to treat viral infections and skin cancers in immunocompetent and immunosuppressed patients. On the other hand, the topical immunosuppressive agents tacrolimus and pimecrolimus have been used with great success in the treatment of chronic inflammatory diseases in children and adults. The introduction of this new class of drugs (i.e. Calcineurin inhibitors) marked the beginning of the post-cortisone era in clinical dermatology. Toll-like receptor agonists and calcineurin antagonists will supplement corticosteroids to improve specific dermatological therapy. Topical immunotherapy with both immunostimulatory and immunosuppressive agents show potential for effective and patient-friendly treatment of inflammatory, infectious and neoplastic skin diseases. Long-term evaluation will define the tolerability and the safety profile. PMID- 12908455 TI - [Sun protection. Correction of an error]. PMID- 12908456 TI - Self-expandable metal mesh stents for common bile duct stenosis in chronic pancreatitis: retrospective evaluation of long-term follow-up and clinical outcome pilot study. AB - BACKGROUND: Symptomatic common bile duct strictures are frequently seen in the course of severe chronic pancreatitis with a reported incidence of 10-30%. The exact role of endoscopic stenting with metal prostheses as definitive treatment has not yet been clearly defined. METHODS: Six patients with symptomatic distal common bile duct stricture in the setting of chronic pancreatitis were treated with a self-expandable metal stent in an attempt to achieve permanent drainage. All cases were approached non-operatively and underwent prior unsuccessful treatment with plastic prostheses. RESULTS: The patients received a self expandable metal Wall stent after a median follow-up of 14 months of plastic stenting. The median follow-up time was 58 months and the stents remained open for a median time of 20 months, respectively. Overall, in 2 of 6 cases the metal stent therapy was successful while 4 of 6 patients required additional endoscopic procedures. CONCLUSIONS: Self-expandable metal stents could be useful and adequate in patients who suffer from symptomatic obstructive jaundice due to chronic pancreatitis. However, they provide acceptable dilation of the common bile duct for only a limited period of time. Overall, endoscopic stenting with self-expandable metal stents seems to be an advantageous treatment only for a subgroup of patients who are unfavorable candidates for surgical drainage. To clarify the role of the endoscopic approach, prospective trials with homogeneous and larger patient numbers are necessarily in the future. PMID- 12908457 TI - Pneumococcal vaccination for cochlear implant candidates and recipients: updated recommendations of the Advisory Committee on Immunization Practices. AB - In October 2002, CDC recommended that all persons with cochlear implants receive age-appropriate pneumococcal vaccination with 7-valent pneumococcal conjugate vaccine (PCV7) (Prevnar), 23-valent pneumococcal polysaccharide vaccine (PPV23) (Pneumovax), or both according to the Advisory Committee on Immunization Practices (ACIP) schedules for persons at high risk. CDC issued these recommendations on the basis of preliminary data suggesting an increased risk for pneumococcal meningitis in persons with cochlear implants. Findings of a recent investigation by CDC, the Food and Drug Administration (FDA), and state health departments support this recommendation. Children aged <6 years with a cochlear implant had a substantially greater risk for having pneumococcal meningitis, compared with children in the general U.S. population of the same age. Some children who are candidates for cochlear implants have pre-existing anatomic factors that might contribute to an increased risk for meningitis; however, the recent study was not designed to assess this association. PMID- 12908458 TI - Binge eating as a phenotype of melanocortin 4 receptor gene mutations. PMID- 12908459 TI - Binge eating as a phenotype of melanocortin 4 receptor gene mutations. PMID- 12908460 TI - Mammographic screening for breast cancer. PMID- 12908461 TI - Mammographic screening for breast cancer. PMID- 12908462 TI - Mammographic screening for breast cancer. PMID- 12908463 TI - Mammographic screening for breast cancer. PMID- 12908464 TI - Skin cancers after organ transplantation. PMID- 12908466 TI - With a little help from your friends... PMID- 12908465 TI - Skin cancers after organ transplantation. PMID- 12908467 TI - Oncology's trials. PMID- 12908468 TI - Bortezomib. PMID- 12908469 TI - Positioning the patient for abdominoperineal excision of the rectum (APER). PMID- 12908470 TI - The open abdomen--a simple cost-effective technique for laparostomy management. PMID- 12908471 TI - A cork in a bottle--a simple technique for removal of a rectal foreign body. PMID- 12908473 TI - The flexor hallucis longus tendon transfer for chronic tendo-achilles ruptures revisited. PMID- 12908474 TI - Use of the Shirodkar suture in shoulder surgery. PMID- 12908472 TI - A simple way to retrieve the gallbladder in '5-mm' laparoscopic cholecystectomy. PMID- 12908475 TI - Use of adrenaline and bupivacaine to reduce bleeding and pain following harvesting of bone graft. PMID- 12908476 TI - An aid to external fixator application. PMID- 12908477 TI - An introduction to legal thought: four approaches to law and to the allocation of body parts. PMID- 12908478 TI - When life is an injury: an economic approach to wrongful life lawsuits. PMID- 12908479 TI - The formulary files. HMI, Medicaid formularies exhibit surprisingly small differences. PMID- 12908480 TI - The timing of third molar removal in patients undergoing a bilateral sagittal split osteotomy. PMID- 12908481 TI - Emerging trends in trauma care. PMID- 12908482 TI - An eye for the ladies. PMID- 12908483 TI - Wireless capsule endoscopy in the diagnosis of small bowel Crohn's disease. PMID- 12908484 TI - Use with caution: 6MP in pregnancy. PMID- 12908485 TI - Antibodies to "antibodies to tumor necrosis factor" and infusion reactions: is this infliximab's Achille's heel? PMID- 12908486 TI - [Health and power: the political emergence of AIDS/HIV in Brazil]. AB - This analysis of the construction of political responses to AIDS in Brazil is grounded on a study of the epidemic since its first appearance, against a backdrop formed by the recent history of Brazilian health policy, the period's emerging social movements, and data on AIDS/HIV evolution. Together with period documents, oral sources constitute a fundamental part of the research. The article sees official responses to AIDS from the time of the disease's political emergence in Brazil as evolving through different stages and eventually -- after some problems and not without certain limitations--becoming characterized first and foremost by society's participation. PMID- 12908487 TI - [Economics, politics, and public health in Porfirian Mexico (1876-1910)]. AB - The article examines the scientific, political, and economic elements that permitted the birth of modern public health in Mexico under the Porfirio Diaz administration (1876-1910). Firstly, a portion of Mexican physicians were open to the discoveries of microbiology, immunology, and epidemiology. Secondly, the State's growing concentration of power in public health matters ran parallel to its concentration of disciplinary political power and enabled this new knowledge to be placed at the service of collective health problem prevention. Lastly, both imperialism and the Porfirian elite needed to protect their business interests. The article evaluates public health achievements and limitations during the Porfirian period, abruptly interrupted by the revolution begun in 1910. PMID- 12908488 TI - [Medical value of humic acid]. PMID- 12908489 TI - James McCubrey: a member of The Editorial Academy of The International Journal of Oncology. PMID- 12908490 TI - Drug-related taste disturbances. AB - Ageusia and dysgeusia can be caused by many systemically administered drugs. PMID- 12908491 TI - Coxibs and anaphylactic shock. PMID- 12908493 TI - Paroxetine withdrawal syndrome. PMID- 12908492 TI - Pneumonitis and non steroidal antiandrogens. AB - Some serious cases with bicalutamide, flutamide and nilutamide. PMID- 12908494 TI - Kava: first suspended, now prohibited. PMID- 12908495 TI - Excess mortality with salmeterol as single-agent therapy. PMID- 12908496 TI - Fatal pneumopathy and gefitinib. PMID- 12908497 TI - Statins: new data in secondary prevention and diabetes. Pravastatin and simvastatin are the best-assessed statins. AB - The efficacy of pravastatin and simvastatin was first shown several years ago in patients with coronary heart disease. Other trials have since been published. In the HPS trial, which studied patients with coronary heart disease, other cardiovascular conditions, or diabetes, simvastatin significantly reduced the risk of death, coronary events and stroke when compared with placebo. In the ALLHAT-LLT trial, in patients with treated hypertension, pravastatin did not reduce overall mortality. In the PROSPER trial, in patients aged over 70 with cardiovascular disease or cardiovascular risk factors, pravastatin reduced the incidence of coronary events relative to placebo, but did not reduce overall mortality. Pharmacovigilance studies suggest there is no difference between these four statins in terms of their potential to cause rhabdomyolysis. Taken together, these trials show that statin use can be extended to patients with levels of LDL cholesterol over 2.4 mmol/l (0.9 g/l) if they have coronary heart disease (and no hypercholesterolaemia), a history of ischaemic stroke, or lower-limb arterial disease. Statins can also be prescribed for diabetic patients with no signs of cardiovascular disease but whose LDL-cholesterol exceeds 3.4 mmol/l (1.3 g/l). Clinical trial data support the use of pravastatin or simvastatin in these situations, at a dose of 20 or 40 mg daily. Plasma creatine phosphokinase assay should be done if muscle symptoms occur or if the patient has a particular risk of rhabdomyolysis. PMID- 12908498 TI - Antibiotics in children with acute otitis media? AB - Acute otitis media is defined as eardrum inflammation associated with middle-ear effusion and acute signs and symptoms. Viruses are the primary pathogens, and bacterial superinfection is common. Pneumococci and Haemophilus influenzae are the most frequently isolated bacterial pathogens. Recovery is usually rapid, though somewhat slower in children under 2 years of age. The only benefits of antibacterial chemotherapy are a marginal reduction in the duration of pain and use of analgesics. Antibacterial therapy has no effect on the risk of relapse, medium term auditory outcome in children, or complications. It does, however, increase the risk of becoming a carrier of resistant strains PMID- 12908500 TI - NSAID interactions with intrauterine devices? PMID- 12908499 TI - Transparency of EU medicines agencies. PMID- 12908501 TI - Heparin prophylaxis for a leg cast? PMID- 12908502 TI - [Advances in the study and development of medical uses of sericulture and mulberry resources]. PMID- 12908503 TI - The neuropsychology of grammatical gender: an introduction. PMID- 12908505 TI - NIH study supports diluting smallpox vaccine stockpile. PMID- 12908504 TI - Is there a future for vaccines? PMID- 12908506 TI - Edible AIDS vaccine milestone. PMID- 12908507 TI - Use of AAV vectors in AIDS vaccine. PMID- 12908508 TI - Inactivated hepatitis A vaccine: immunogenicity, efficacy, safety and review of official recommendations for use. AB - There is 10 years of marketing experience with the hepatitis A vaccine Havrix. It is highly immunogenic, provides lasting protection in healthy individuals and generates protective levels of antibodies in patients with chronic liver disease or impaired immunity. Postmarketing surveillance data have confirmed the outstanding safety profile of the vaccine. The timing of the booster dose is not critical to effectiveness, which has advantages for the protection of travelers to regions of high endemicity. The vaccine is effective in curbing outbreaks of hepatitis A and also when administered postexposure, due to rapid seroconversion and the long incubation period of the disease. In intermediate endemic regions, an epidemiological shift in hepatitis A infection has driven the development of universal preventive strategies to be added to the targeting of at-risk groups. Existing official recommendations and future directions for vaccine use are reviewed. PMID- 12908509 TI - Vaccination to prevent type 1 diabetes. AB - Immune-based interventions to treat or prevent autoimmune disorders are opening up entirely novel therapeutic avenues. Presently, systemic immune modulators such as antibodies to cytokines or cell-surface molecules (i.e., costimulators) are undergoing clinical testing or are already in use. The next generation of interventions will function in an antigen-specific way. These strategies are more difficult to design, because successful immunization or vaccination has to be monitored on an individual basis and dose/route/timing considerations play an essential role. Both will be discussed in this chapter and suitable combinatorial approaches will be suggested. PMID- 12908510 TI - Breast cancer vaccines: current research and future prospects. AB - Although the immune system can clearly recognize cancer cells, there is little evidence that it does so to any effective consequence in patients with advanced breast cancer. The possibility of effective control of micrometastatic disease remains but is unproven. The challenge for the clinician is to find ways to enhance the ability of a patient's own immune system to recognize and effectively target malignant cells. It is clear that there is no shortage of target antigens for immune cell recognition of breast cancer but it is less certain whether the route to an effective therapeutic strategy lies with enhanced recognition of an individual target or a more general route to enhanced immune cell activation. PMID- 12908511 TI - Immunomodulation with antibodies: clinical application in ovarian cancer and other malignancies. AB - This review identifies the role of antibodies in the field of therapeutic cancer vaccines. Ovarian cancer is used as a model to review advances in therapeutic vaccine development with a focus on antibodies as immunomodulators and antigen mimetics, highlighting research on B43.13 and ACA125. The interaction of biological immunomodulation and chemotherapy is discussed. Requirements of antigen processing and recent advances in the field of dendritic cell biology are critical to current understanding of potent immune response induction. Future directions including use of growth factors, adjuvants and cellular therapies to enhance effects may potentiate observed effects and provide clues for application in many cancers and beyond oncology. PMID- 12908512 TI - CEA-based vaccines. AB - Carcinoembryonic antigen (CEA), the first tumor-associated antigen to be described, was cloned in 1987 and is expressed on nearly 50% of all human tumors. The identification of T-cell specific epitopes within the coding region of the CEA protein has led to the development of various vaccine strategies that target CEA and CEA-expressing tumors. These vaccines have shown evidence of therapeutic effectiveness in animal models and are being evaluated in early phase clinical trials. Although trials are not designed to elucidate clinical responses, they have provided important information about the ability of individual vaccines to induce CEA-specific immune responses through the use of newer in vitro monitoring assays. Continued clinical testing in patients with less advanced disease and the administration of vaccines in combination with other standard therapy will help define the role of CEA-based vaccines in the treatment and prevention of human cancer. PMID- 12908513 TI - Pneumococcal conjugate vaccine: economic issues of the introduction of a new childhood vaccine. AB - In February 2000, a pneumococcal conjugate vaccine was licensed for use in the USA. This vaccine has been shown to be effective in reducing pneumococcal disease, and has been recommended for universal use in infants. However, pneumococcal conjugate vaccine is by far the most expensive child vaccine series routinely administered in the USA, alone accounting for over 40% of the total purchase price of vaccines for the recommended childhood schedule. This article reviews the existing efficacy and economic studies of pneumococcal conjugate vaccine and discusses the process by which routine use of pneumococcal conjugate vaccine was introduced and the role economic analysis played in that process. Some of the scientific and funding issues relating to its use in both the industrialized and developing world are also discussed. PMID- 12908515 TI - Technological advances to increase immunogenicity of DNA vaccines. AB - The latest clinical data obtained with DNA vaccines against HIV and malaria have shown promise, but it is clear that when DNA vaccines are compared with other vaccine vector delivery systems, there is still room for improvement. Further development is more than possible, based on the wealth of information accumulating on methods and approaches to increase immunogenicity of DNA vaccines. Thus, the goal of this review is to summarize some of the latest technological advances to increase immunogenicity of DNA vaccines administered by the im. and id. routes. By means of examples, the review will be intended to focus only on recent developments reported in the last 2 years and likely to go towards the improvement of mucosal, humoral and cellular immune responses mostly against cancer and infectious disease antigens. PMID- 12908514 TI - Meningococcal vaccines. AB - The successful introduction of a protein-polysaccharide conjugate vaccine against serogroup C meningococci into the UK infant immunization schedule, in combination with a catch-up campaign for individuals less than 18 years of age, has seen virtually all group C disease eliminated in childhood. From being a devastating disease with a very high mortality, the possibility of eradicating invasive meningococcal disease now seems eminently feasible. Since similar technology is likely to facilitate prevention of disease caused be serogroup A, Y and W135 meningococci, the major hurdle in achieving the goal of eradication is development of a safe and immunogenic vaccine against serogroup B infections. Outer membrane vesicle vaccines remain in development and further trials are anticipated. Through the recent availability of the meningococcal genome sequence, many new vaccine candidates are being identified and there is increasing optimism that a solution to the problem can be found. PMID- 12908516 TI - Open system gene expression profiling and identification of novel genes for targeted vaccine delivery. AB - Ongoing development of vaccines is limited by the techniques used to identify routes to immunization. Using open system gene expression profiling, researchers can now directly identify receptors used by M-cells in the transport of antigens to the mucosal immune system. These receptors will be ideal for targeted delivery of new synthetic vaccines. In addition, gene expression profiling will provide important new information on the biology of the M-cell and mucosal immune mechanisms. PMID- 12908517 TI - Virus-like particles: a new family of delivery systems. AB - The development of antiviral vaccines has almost exclusively been based on live attenuated vaccines up until now. However, the efficacy of HBsAg particles as an antiHBV vaccine has clearly demonstrated that protective antiviral immunity can be achieved by other strategies. Virus-like particles formed by structural proteins were proven to be highly immunogenic and capable of inducing protective immunity against various viral infections in preclinical studies. Clinical trials using virus-like particles confirmed their safety and immunogenicity. Moreover, chimeric virus-like particles carrying foreign peptidic sequences were shown to elicit potent B- and T-cell responses. Virus-like particles formed by a fusion protein between the HBsAg and the circumsporozoite surface protein are safe and immunogenic in volunteers and induce a partial protection against natural Plasmodium falciparum infection. PMID- 12908518 TI - Montanide ISA 720 and 51: a new generation of water in oil emulsions as adjuvants for human vaccines. AB - The development of adjuvants will represent a major challenge for this century. Indeed the need for safer vaccines leads to the development of a new generation of antigens like synthetic peptide, recombinant proteins or even vectored DNA. However, this is to the detriment of their immunogenicity. The addition of adjuvant is becomes necessary to enhance immune responses and improve vaccine potency. However, adjuvants can be responsible for the apparition of secondary reactions and they must be adapted according to various criteria such as the route of immunization, the type of the immune response, the duration of immunity, or the quality of the antigen, in order to get the best balance between efficacy and safety. PMID- 12908519 TI - Disappearance of CD21-positive follicular dendritic cells preceding the transformation of follicular lymphoma: immunohistological study of the transformation using CD21, p53, Ki-67, and P-glycoprotein. AB - Some follicular lymphomas histologically transform into diffuse aggressive lymphomas, the prognosis of which is poor. There are, however, no reliable histological criteria for predicting which cases will later undergo such transformation. In low-grade B-cell lymphomas, follicular dendritic cells form dense mesh-like networks that contain accumulating neoplastic B-cells. These are rare in high-grade lymphomas. We immunohistochemically analyzed CD21-positive follicular dendritic cells in 32 follicular lymphomas, including 3 transformed lymphomas, in addition to immunohistological study using P-glycoprotein, p53, and Ki-67. We found that the mesh-like networks in follicles are more clearly defined in low-grade lymphomas than in high-grade lymphomas (p = 0.015). Neoplastic follicles in 2 transformed lymphomas lost the networks of follicular dendritic cells before transformation despite the existence of morphologically clear follicles. This differed from the non-transformed cases of the same cytological grades. Prognosis was statistically better for patients with low-grade tumor than for those with high-grade tumor (p = 0.026), and there was a trend toward poorer survival among CD21-negative cases (p = 0.186). P-glycoprotein, p53, and Ki-67 expressions did not provide sufficient information to predict the transformation of follicular lymphoma. The presence of CD21-positive follicular dendritic cells in neoplastic follicles might help predict the potential of follicular lymphoma to transform to diffuse large B-cell lymphoma. PMID- 12908520 TI - Characterization of human papillomavirus infection, P53 and Ki-67 expression in cervix cancer of Mozambican women. AB - In this study, we aimed at evaluating the distribution of HPV types and the expression of P53 and Ki-67 in cervix carcinomas of Mozambican women. Fourty seven invasive carcinomas, 10 CIN III, and 10 normal cervix were studied. P53 and Ki-67 expression was examined immunohistochemically. HPV infection and HPV types were detected by PCR (GP5+/bio-GP6+) and enzyme-immunoassay, respectively. Expression of P53 and Ki-67 and detection of HPV were as follows: normal cervix- 0%, 10%, and 0%, respectively; CIN III--10%, 0%, and 100%, respectively; invasive carcinomas--50%, 55.5%, and 70%, respectively. HPV 16 was identified in 54% of invasive carcinomas, HPV 31, 33, 35, and 45 in 23%, "unidentified" HPV in 19%, and HPV 18 in 4% of invasive carcinomas. No significant associations were observed between P53 expression, Ki-67 expression, and HPV infection. In conclusion, we observed a high frequency of HPV infection in CIN III lesions and invasive carcinomas from Mozambican women, with HPV 16 representing the most frequent viral type. HPV status was not related to P53 and Ki-67 expression. Both P53 and Ki-67 are associated with invasive cervix carcinomas, mainly of the squamous keratinizing histotype. PMID- 12908522 TI - Expression of p53 and mdm2 and their significance in recurrence of superficial bladder cancer. AB - The purpose of this study is to evaluate the expression of p53 and mdm2 and to determine whether they may be used as additional predictors of recurrence in superficial transitional cell carcinoma of the bladder. Paraffin sections of 80 patients with superficial transitional cell carcinoma of the bladder, who were treated with transurethral resection, were stained with p53 and mdm2 antibodies using the standard avidin biotin immunoperoxidase method. Nuclear staining for both p53 and mdm2 was calculated as the percentage of labeled nuclei out of a total number of tumor cells counted. The percentage of p53- and mdm2-positive cells showed a significant relationship with tumor grade and recurrence (p = 0.002 and p = 0.016; p = 0.01 and p = 0.003, respectively). In addition, a weak inverse relationship was found between p53 and mdm2 values (r = -0.184). p53 and mdm2 reactivities are valuable parameters in predicting recurrence in superficial bladder cancer. Thus, mdm2 expression appears to play a role in predicting biologic behavior in superficial transitional carcinoma of the bladder. PMID- 12908521 TI - Deletion mapping of chromosome 1 in early onset and late onset breast tumors--a comparative study in eastern India. AB - Younger women exhibit more aggressive pathologic features of breast cancer (BC) compared to their older counterparts. Young age has been shown to be an independent predictor of adverse prognosis. These findings have raised the question of whether these differences are also present at the genetic level. Twenty-five early onset (age < or = 40 years) tumors including 4 bilateral tumors, and 26 late onset (>40 years) breast tumors, including 2 bilateral tumors, were examined for loss of heterozygosity (LOH) at chromosome 1 using 11 polymorphic microsatellite markers. A comparative study revealed high frequencies of LOH in chr. 1p36 (61%), 1p31.3 (40%), 1p21.3 (50%) and 1q22-23.2 (56%) in a younger group, and chr. 1p36 (46%), 1p34.2 (48%), and 1q22-23.2 (52%) in an older group. These differences in LOH frequency in these two age groups were significant for chr. 1p21.3 (p = 0.025) only. These data suggest that the deletion pattern in early onset breast tumors is not fully identical to late onset breast tumors. Similar differential deletion patterns of LOH in the 5 highly deleted regions were seen in premenopausal and postmenopausal groups. An association was seen between LOH at chr. 1p34.2 and chr. 1q22-23.2 and higher grade of the tumors in older women. Among the highly deleted regions, the deletion at chr. 1p36 was found to occur early in both groups because of common allelic loss in the bilateral tumors. PMID- 12908523 TI - Magnesium whitlockite, a calcium phosphate crystal of special interest in pathology. AB - Whitlockite (in fact magnesium whitlockite) is a calcium orthophosphate crystal in which, in biological conditions, magnesium is partly substituted for calcium. Identified in X-ray or electron diffraction patterns, it occurs in physiological or pathological conditions at extra or intratissular sites, mainly in tissues of non-epithelial origin. In a range of pathological calcifications investigated by X-ray diffraction, we noted that whitlockite appeared to be frequently associated with apatite, particularly in "dystrophic calcifications" of tuberculous origin. These personal observations could be correlated with documented data in oral pathology (dental calculus, salivary stones, and dental caries). Whitlockite deposits have also been reported in non-infectious conditions, such as in aortic media, cartilage, and bone tissue. Whereas the formation of both apatite and magnesium whitlockite appears to be caused by the binding of their constituting ions with proteolipids, magnesium inhibits apatite originating from amorphous calcium phosphate to the benefit of whitlockite formation. Possibly, the development of magnesium whitlockite may provide an interesting marker for magnesium metabolism. Further studies linking histology to crystallography might relate the crystal to issues, such as tuberculous calcifications or diseases of bone tissue, and might be useful for potential diagnostic orientation. PMID- 12908524 TI - Detection of apoptoses in gastro-intestinal graft-versus-host disease and cytomegalovirus colitis by a commercially available antibody against caspase-3. AB - The common pathway of any apoptotic cascade leads to the activation of the so called execution caspases, particularly caspase-3 (CSP3). The question of whether immunohistochemical (IHC) detection of activated CSP3 might be useful for routine purposes still needs to be clarified. We analyzed apoptoses in gastrointestinal graft-versus-host disease (GvHD) and cytomegalovirus (CMV) colitis using a commercially available polyclonal antibody against activated human CSP3. In GvHD samples obtained from the colon, apoptoses detected by CSP3 varied between 11 and 43/40 crypts, and in esophageal specimens between 21 and 40/1.5 mm squamous epithelium basal length. This count was correlated with the apoptotic count assessed on hematoxylin- and eosin (H&E)-stained slides. A perfect concordance for those counting between 30 and 40 apoptoses/40 crypts or 1.5 mm squamous epithelium basal length was detected, whereas cases with low apoptotic counts on the H&E stained slides showed a 2 to 3 fold greater number of stained nuclei as assessed by CSP3 staining. In CMV-colitis, although the number of exploding crypt cells was 8-13/40 crypts, only 1-2 nuclei/40 crypts and almost all cells with typical nuclear inclusions stained positively. The presence of CMV can be easily detected on H&E- or IHC-stained slides, while masked GvHD by an overlying CMV colitis might remain unrecognized. Staining for CSP3 may be helpful in distinguishing these two conditions, as apoptotic count would be excessive in GvHD. PMID- 12908525 TI - Culturing mouse peritoneal mesothelial cells. AB - Obtaining normal cells has become increasingly important for use in comparative genetic analytical techniques to examine alterations in gene expression during transformation and progression into malignancy. Normal mesothelial cells are not currently available in cell banks and are essential for comparison of genetic expression analysis in current mouse mesothelioma models. The purpose of this investigation was to extract normal mouse peritoneal mesothelial cells using minimal culture techniques to obtain sufficient cells for gene expression analysis. Mesothelial cells were collected from the mouse peritoneal cavity in vivo with minimal contamination of lymphocytes and macrophages. The cells were cultured for approximately eight days until they were just confluent and retained normal mesothelial phenotype and cytokeratin immunoperoxidase staining. PMID- 12908526 TI - Carcinosarcoma of the colon: report of a rare tumor. AB - Carcinosarcoma is a rare tumor that shows both epithelial and stromal malignant differentiation. Most reported cases of carcinosarcoma affect the female genital tract (and are called malignant mixed mullerian tumors), but there are also some isolated reports of cases affecting the lung and the head and neck area. Carcinosarcomas only rarely affect the gastrointestinal tract, mainly the esophagus. To the best of our knowledge, only eight cases of carcinosarcoma of the colon have been reported to date. For some lesions, the term 'sarcomatoid carcinoma' is preferred to 'carcinosarcoma', as both stromal and epithelial cells have shown cytokeratin expression on immunohistochemistry. The expression 'carcinosarcoma' should be applied only to those lesions, the stromal elements of which do not express epithelial markers. We report a new case of carcinosarcoma affecting the left colon. The most unique feature of this tumor is that it shows chondro-and osteosarcomatous differentiation, a feature that has been described previously in only one colonic carcinosarcoma. We discuss the histopathological and immunohistochemical features of this lesion as well as its possible histogenesis. PMID- 12908527 TI - Tumor embolism as a cause of an unexpected death: a case report. AB - The primary causes of deaths for individuals with rare cancers can be difficult to diagnose clinically. Often, the symptoms implicate a variety of factors, and an autopsy is thus required to obtain the correct diagnosis. This study analyzes the death of a 45-year-old woman who reportedly died from an acute pulmonary dysfunction. The patient had been treated with antibiotics for three months for intractable pneumonia. Suspicious coin lesions detected by chest X-ray prompted a clinical clarification; however, no final diagnosis was made. The autopsy revealed a bulky thyroid tumor with venous invasion, leading to a massive pulmonary tumor embolism. Furthermore, microscopy identified the tumor as a rare pleomorphic myxoid sarcoma. Thus, the patient died of a large pulmonary tumor embolism originating from this rare sarcoma, and not of acute pulmonary dysfunction of any other means. PMID- 12908528 TI - Increased incidence and severity of retinopathy of prematurity in developing nations. PMID- 12908529 TI - Combined resection and anterior transposition of the inferior oblique muscle for the treatment of moderate to large dissociated vertical deviation associated with inferior oblique muscle overaction. PMID- 12908530 TI - Trends in the management of retinoblastoma: evaluation of 1,196 consecutive eyes during 1974 to 2001. PMID- 12908531 TI - Laser treatment of zone I prethreshold and stage 3 threshold retinopathy of prematurity. AB - PURPOSE: To analyze the structural outcomes of zone I prethreshold and stage 3 threshold retinopathy of prematurity following laser ablation. PATIENTS AND METHODS: This nonrandomized observational case series evaluated 71 eyes of 36 infants who weighed less than 1,250 g at birth. Infants were treated at either prethreshold or threshold severity. The structural outcomes were compared using Fisher's exact test. RESULTS: The structural outcome was favorable in 96% and 80% of the prethreshold and threshold groups, respectively (P = .069). When an unfavorable outcome was defined as dragging of the temporal vessels, the difference was significant (P = .0257). CONCLUSION: Early treatment of zone I disease may result in a better structural outcome. PMID- 12908533 TI - Comparison of cosmetic results in frontalis sling operations: the eyelid crease incision versus the supralash stab incision. AB - PURPOSE: In addition to ptosis, the lack of eyelid crease is one of the main cosmetic problems of patients with poor levator function. In this study, we aimed to evaluate the effect of the incision site on ptosis correction and eyelid crease formation in frontalis sling operations. MATERIALS AND METHODS: Photographs and charts of 27 patients (45 eyelids) who underwent frontalis sling operations were retrospectively reviewed. All patients had congenital ptosis with poor levator function ranging from 0 to 4 mm. In group I (15 eyelids), 3 supralash eyelid skin incisions were performed and the sling material was threaded along the tarsal surface. In group 11 (30 eyelids), the eyelid crease approach was used with the suturing of the sling material directly to the tarsus. RESULTS: Although lid contour and lid crease symmetry were good in 46.6% of the cases in group I, eyelid crease formation was not satisfactory in any of the cases. Lid contour and lid crease symmetry were good in 76.6% of the cases in group II. CONCLUSION: Our results indicated that the eyelid crease approach provided better functional and cosmetic results than did the supralash stab incision in frontalis sling operations. The passage of the sling material behind the orbital septum by direct visualization in the eyelid crease approach is one of the main factors affecting the surgical success of the frontalis sling operation. PMID- 12908532 TI - Incidence and severity of retinopathy of prematurity in Vietnam, a developing middle-income country. AB - BACKGROUND: Although retinopathy of prematurity (ROP) is a leading cause of childhood blindness, its impact in lower income countries is not well documented. The World Health Organization has proclaimed that infants at risk for ROP should have screening eye examinations and access to treatment. PATIENTS AND METHODS: A prospective study was conducted from January 1 through December 31, 2001, at Tu Du Hospital in Ho Chi Minh City for premature infants who weighed 1,500 g or less at birth or were 33 gestational weeks or younger. Serial examinations were used to classify ROP, and treatment outcomes were noted. RESULTS: Two hundred twenty five consecutive infants were included in the data analysis. Birth weights ranged from 900 to 2,000 g (mean, 1,512 g). Gestational ages ranged from 26 to 36 weeks (mean, 31 weeks). ROP was present in 103 (45.8%) of the 225 infants. In infants who weighed 1,250 g or less at birth, the ROP rate was 81.2% (26 of 32 infants). Threshold ROP was present in 9.3% of the 225 infants but in 25% of the 32 infants. Twenty-four eyes received treatment, whereas 16 lacked the family resources. Of the 24 treated eyes, 18 (75%) had a favorable outcome. Of the 16 untreated eyes, only 3 had a favorable outcome. CONCLUSIONS: ROP incidence is high in Vietnam, similar to that in the United States. However, larger, older infants are at risk in Vietnam and the rate of severe ROP seems to be higher. This necessitates an ROP screening paradigm different from that currently used in the United States. PMID- 12908534 TI - Surgery on the nonparetic eye for oculomotor palsy with aberrant regeneration. PMID- 12908535 TI - Prenatal diagnosis of retinoblastoma. PMID- 12908536 TI - Tonic pupil due to supranuclear third nerve paresis. PMID- 12908537 TI - Craniometaphyseal dysplasia: ophthalmic features and management. PMID- 12908538 TI - Isolated bilateral optic neuropathy in acute disseminated encephalomyelitis. PMID- 12908539 TI - Electroretinographic findings in a full-term newborn with retinal hemorrhages. PMID- 12908540 TI - Panuveitis due to acquired rubella and isolation of rubella virus from the aqueous humor. PMID- 12908541 TI - Lisch nodule asymmetry in a patient with neurofibromatosis type 1. PMID- 12908542 TI - Transient monocular diplopia resulting from the treatment of amblyopia. PMID- 12908543 TI - Pilomatrixoma of the eyelid simulating a chalazion. PMID- 12908544 TI - Driving the cell cycle to cancer. AB - Cell cycle progression requires the co-ordinated activation of several kinases, some of which are activated upon the binding of a cyclin subunit. At least four of these so-called cyclin-dependent kinases, namely Cdk4, Cdk6, Cdk2 and Cdk1, have specific roles at particular stages of the cell cycle, including passage through the various cell cycle transitions and the response to specific checkpoints. Not surprisingly, most human tumors carry mutations that deregulate at least one of these kinases. To analyze their specific role in vivo, we are generating strains of gene-targeted mice carrying either activated or defective alleles of these Cdks. As an example, Cdk4 expression appears to be expendable in most cell types since mice lacking Cdk4 are viable. Yet, Cdk4 mutant mice are smaller in size and infertile (only partial infertility in males). In addition, Cdk4 defective mice develop insulin dependent diabetes early in life. However, the importance of these Cdks in tumor cell cycles is underscored by the phenotype of knock in mice where the normal Cdk4 gene has been replaced by a Cdk4 R24C (insensitive to INK inhibitors) mutant. These animals develop a wide spectrum of spontaneous tumors and are highly susceptible to specific carcinogenic treatments. These models are being used now to understand how deregulation of these Cdks leads to cancer development and will be a valuable tool to design and validate new therapeutic strategies against tumour development. PMID- 12908545 TI - Proliferation: the cell cycle. PMID- 12908546 TI - Molecular analysis of gene expression in tumor pathology. AB - Human cancers are diverse in their pathology and responsiveness to clinical treatment. This diversity is at least in part due to variations in cellular gene expression programs. Although the analyis of proteins--the key players in cells and potential drug targets--is advancing rapidly, there are situations in which the analysis of RNA rather than proteins can provide valuable information for the diagnosis of cancer. These situations include absense of an antibody for the protein of interest, expression of functionally defective proteins, expressed small nucleotide polymorphisms (SNPs), analysis of alternatively or abnormally spliced molecules, and functional analysis of splice site mutations. In this chapter we will focus on the analysis of RNA from clinical samples and will summarize how gene expression studies on the RNA level using a variety of new tools can be useful for discovering new classes of tumors, for predicting clinical outcome or therapy response, and for designing novel personalized clinical interventions that can not be achieved with histology alone. PMID- 12908547 TI - Ewing tumor biology: perspectives for innovative treatment approaches. AB - The Ewing's sarcoma family of tumors (EFT) is a group of malignancies affecting bone and soft tissue in adolescents. It is characterized by a unique gene rearrangement between the EWS gene and an ets transcription factor gene. EFT can be cured with conventional multi modal treatment, however, about 40% of patients still succumb to the disease. Relapses can be observed more than 5 years after the end of primary treatment suggesting persistence of minimal residual disease (MRD). Due to the still enigmatic nature of EFT histogenesis the phenotype of EFT stem cells and of dormant tumor cells remains unknown. The most frequent fusion product associated with EFT, EWS-FLI1, is the founding member of a whole class of similarly structured chimeric proteins associated with a variety of human sarcomas and also specific leukemias. The corresponding gene rearrangement constitutes a rate limiting step in oncogenesis as implied by the high association of EFT with EWS-ETS fusions, strong selective pressure for maintenance of a correct reading frame in the tumors, and by experimental data confirming the transforming and tumorigenic potential of EWS-FLI1. Understanding the biology of EWS-ETS gene fusions and its interplay with essential cellular pathways regulating cell growth, apoptosis, differentiation, genomic integrity, and treatment resistance may unravel specifically vulnerable sites for therapeutic targeting. This review summarizes the current knowledge about the EWS FLI1 pathway in EFT and provides some ideas as to how this knowledge may be translated into innovative treatment approaches. PMID- 12908548 TI - Cancer epigenetics: DNA methylation and chromatin alterations in human cancer. AB - Aberrations in the DNA methylation patterns are nowadays recognized as a hallmark of human cancer. One of the most characteristic changes is the hypermethylation of CpG islands of tumor suppressor genes associated with their transcriptional silencing. The target genes are distributed in all cellular pathways (apoptosis, DNA repair, cell cycle, cell adherence, etc.). They are "classical" tumor suppressor genes with associated familial cancers (BRCA1, hMLH1, p16INK4a, VHL, etc.) and putative new tumor suppressor genes which loss may contribute to the transformed phenotype (MGMT, p14ARF, GSTP1, RARB2, etc.). A tumor-type specific profile of CpG island hypermethylation exist in human cancer that allows the use of these aberrantly hypermethylated loci as biomarkers of the malignant disease. The eruption of new technologies for the careful study of the DNA methylation patterns, and their genetic partners in accomplishing gene silencing, it may also provide us with new drugs for the epigenetic treatment of human tumors. PMID- 12908549 TI - Molecular analysis of cancer using DNA and protein microarrays. AB - In conclusion, array-based technologies have emerged that contribute to profiling tissues at the genomic, transcriptomic and proteomic levels. Analytical tools are needed to mine the vast amount of data generated. Ultimately the molecular analysis of cancer at a genome and proteome scale will allow better classification of disease and tailored individualized therapy for individual patients. PMID- 12908550 TI - Proteomic approaches to the diagnosis, treatment, and monitoring of cancer. AB - The field of proteomics holds promise for the discovery of new biomarkers for the early detection and diagnosis of disease, molecular targets for therapy and markers for therapeutic efficacy and toxicity. A variety of proteomics approaches may be used to address these goals. Two-dimensional gel electrophoresis (2D-PAGE) is the cornerstone of many discovery-based proteomics studies. Technologies such as laser capture microdissection (LCM) and highly sensitive MS methods are currently being used together to identify greater numbers of lower abundance proteins that are differentially expressed between defined cell populations. Newer technologies such as reverse phase protein arrays will enable the identification and profiling of target pathways in small biopsy specimens. Surface-enhanced laser desorption/ionization time-of-flight (SELDI-TOF) analysis enables the high throughput characterization of lysates from very few tumor cells or body fluids and may be best suited for diagnosis and monitoring of disease. Such technologies are expected to supplement our arsenal of mRNA-based assays, and we believe that in the future, entire cellular networks and not just a single deregulated protein will be the target of therapeutics and that we will soon be able to monitor the status of these pathways in diseased cells before, during and after therapy. PMID- 12908551 TI - Structural basis of tumoral angiogenesis. PMID- 12908552 TI - Matrix metalloproteinases and tumor progression. AB - The matrix metalloproteinases (MMPs) are a family of more than 20 distinct enzymes that are frequently overexpressed in human tumors. Functional studies have shown that MMPs play an important role in the proteolytic destruction of extracellular matrix and basement membranes, thereby facilitating tumor invasion and metastasis. In addition, these enzymes may also be important in other steps of tumor evolution including neoplastic cell proliferation and angiogenesis stimulation. On the basis of the relevance of MMPs in tumor progression, a number of different strategies aimed to block the unwanted activity of these enzymes in cancer have been developed. Unfortunately, most clinical trials with the first series of MMP inhibitors have failed to show clear benefit in patients with advanced cancer. Explanations for this lack of success include the failure to recognize the role of these enzymes in early stages of the disease as well as inadequacy of either the employed inhibitors or the proteases to be targeted. The introduction of novel concepts such as tumor degradome, and global approaches to protease analysis, may facilitate the identification of the relevant MMPs that must be targeted in each individual cancer patient. On the other hand, the finding that MMPs are enzymes whose effects on biologically active substrates can have profound consequences on cell behaviour, suggests that selective inhibition of a limited set of MMPs at early stages of tumor evolution might be much more effective than using wide-spectrum inhibitors active against most family members, and administered to patients at late stages of the disease. Further studies directed to elucidate these questions will be necessary to clarify whether any of the multiple strategies of MMP inhibition may be part of future therapeutic approaches to control tumor progression. PMID- 12908553 TI - Angiogenesis inhibitors and their therapeutic potentials. PMID- 12908554 TI - Mutated tyrosine kinases as therapeutic targets in myeloid leukemias. AB - Tyrosine kinases are commonly mutated and activated in both acute and chronic myeloid leukemias. Here, we review the functions, signaling activities, mechanism of transformation, and therapeutic targeting of two prototypic tyrosine kinase oncogenes, BCR-ABL and FLT3, associated with chronic myeloid leukemia (CML) and acute myeloid leukemia (AML), respectively. BCR-ABL is generated by the Philadelphia chromosome translocation between chromosomes 9 and 22, creating a chimeric oncogene in which the BCR and c-ABL genes are fused. The product of this oncogene, BCR-ABL, has elevated ABL tyrosine kinase activity and transforms hematopoietic cells by exerting a wide variety of biological effects, including reduction in growth factor dependence, enhanced viability, and altered adhesion of chronic myelocytic leukemia (CML) cells. Elevated tyrosine kinase activity of BCR-ABL is critical for activating downstream signalling cascades and for all aspects of transformation, explaining the remarkable clinical efficacy of the tyrosine kinase inhibitor, imatinib mesylate (STI571). By comparison, FLT3 is mutated in about one third of all cases of AML, most often through a mechanism that involves an internal tandem duplication (ITD) of a small number of amino acid residues in the juxtamembrane domain of the receptor. As is the case for BCR ABL, these mutations activate the kinase activity constitutively, activate multiple signaling pathways, and result in an augmentation of proliferation and viability. Transformation by FLT3-ITD can readily be observed in murine models, and FLT3 cooperates with other types of oncogenes to create a fully transformed acute leukemia. FLT3 tyrosine kinase inhibitors are currently being evaluated in clinical trials and may be very useful therapeutic agents in AML. PMID- 12908556 TI - Immune-promoted tumor cell invasion and metastasis. New considerations in cancer therapy. PMID- 12908555 TI - Targeting PDGF receptors in cancer--rationales and proof of concept clinical trials. AB - The platelet-derived growth factors (PDGF) are a pleotrophic family of peptide growth factors that signal through cell surface, tyrosine kinase receptors (PDGFR) and stimulate various cellular functions including growth, proliferation, and differentiation. To date, PDGF expression has been demonstrated in a number of different solid tumors, from glioblastomas to prostate carcinomas. In these various tumor types, the biologic role of PDGF signaling can vary from autocrine stimulation of cancer cell growth to subtler paracrine interactions involving adjacent stroma and vasculature. The tyrosine kinase inhibitor imatinib mesylate (formerly STI571, Gleevec, Novartis Pharmaceuticals Corp, East Hanover, NJ) blocks activity of the Bcr-Abl oncoprotein and the cell surface tyrosine kinase receptor c-Kit, and as such was recently approved for several indications in the treatment on chronic myeloid leukemia and gastrointestinal stromal tumors. In both of these examples the target protein was identified by an oncogenic, activating mutation. Imatinib mesylate is also a potent inhibitor of PDGFR kinase and is currently being evaluated for the treatment of chronic myelomonocytic leukemia and glioblastoma multiforme, based upon evidence in these diseases of activating mutations in PDGFR. However, the PDGF pathway may represent a therapeutic target in other solid tumors in which it is not part of the oncogenic transformation. In order to investigate the potential biologic implications of inhibiting PDGFR in these tumor types, clinical trials that investigate both established clinical endpoints of response and benefit, as well as surrogate endpoints that describe the biologic significance of PDGF inhibition in vivo are needed. PMID- 12908557 TI - Improvements of survival in nine phase II clinical studies with different types of cancer upon anti-tumor vaccination with an autologous tumor cell vaccine modified by virus infection to introduce danger signals. PMID- 12908558 TI - Causation and prevention of solely estrogen-induced oncogenesis: similarities to human ductal breast cancer. AB - Estrogens are intimately involved in the causation of some of the most prevalent cancers afflicting women, particularly, breast, endometrial, cervico-vaginal, and possibly ovarian. Therefore, it has become particularly pertinent to elucidate the molecular changes and mechanisms whereby estrogens elicit their oncogenic actions so that better prevention strategies can be developed. The estrogen induced Syrian hamster tumors of the kidney have emerged as one of the most intensively studied in-vivo models in solely estrogen-induced oncogenesis. An advantage of this model is that the tumors occur in the absence of any intervening morphologic changes, but rather they are the result of the continuous progression of a subset of interstitial stem cells in the kidney leading to tumor formation. Evidence is presented that the origin of these tumors is derived from ectopic "uterine" stem cells, which are responsive to estrogenic hormones. The other animal tumor model studied is the highly sensitive estrogen-induced mammary tumors of female ACI rats. Their steroid receptor and other gene alterations have been delineated. Importantly, a crucial early event in this solely estrogen induced oncogenic process, common to both animal tumor models, is the overexpression and amplification of c-myc and its protein product. Chromosomal instability, in both early and large well-established frank tumors, is another important characteristic found during early E-induced oncogenesis. These features have been shown to be characteristic of human ductal carcinomas in-situ and in primary invasive ductal breast carcinomas. The molecular alterations seen are considered crucial in eliciting estrogen-induced oncogenesis and have established for the first time a direct causal link between estrogen and the induction of chromosomal instability and aneuploidy in these estrogen-associated neoplasms. PMID- 12908559 TI - Cyclooxygenase-2 inhibitors in cancer prevention and treatment. PMID- 12908560 TI - Exosomes for immunotherapy of cancer. AB - Exosomes are 60 to 90 nm membrane vesicles originating from late endosomes and secreted from most hematopoietic and epithelial cells in vitro. B cell derived exosome antigenicity was first reported in 1996 in MHC class II restricted CD4+ T lymphocytes. In 1998, we reported that dendritic cell derived-exosomes are immunogenic in mice leading to tumor rejection. These findings have renewed the interest in exosomes. The current challenge consists in understanding the mechanisms and the physiological relevance of exosomes that could contribute to the design of the optimal exosome based-vaccination. Here, we will focus on the biological features pertaining to dendritic cell- and tumor cell derived-exosomes and will discuss their potential clinical implementation. PMID- 12908561 TI - Breast cancer gene expression analysis--the case for dynamic profiling. AB - Cancer is a complicated disease with each individual tumor exhibiting to a variable degree a set of overlapping phenotypes that include abnormal proliferation and cell survival, genetic instability, tissue invasion, metastasis and angiogenesis. Each of these pathophysiological functions requires abnormal signaling that fails to respect the normal restraints imposed on healthy cells through homeostatic regulation. Understanding of deregulated gene expression is therefore an essential goal for cancer research because unraveling this problem will provide insights into the fundamental nature of cancer as well as provide opportunities for therapeutic intervention. Gene expression profiling is a remarkable technique that addresses this complexity by documenting the expression of thousands of genes at the level of mRNA abundance. From the standpoint of clinical investigation, the initial focus of gene profiling experiments has been on screening primary tumors for markers of poor prognosis. However this is only a first step in demonstrating the utility of this technique. Future advances will focus on understanding the evolution of an organ confined primary cancer to a treatment resistant lethal systemic disease through repeated tumor sampling and analysis, an approach termed in this paper "dynamic profiling". Using breast cancer endocrine therapy as an example our initial approaches to dynamic profiling will be described. PMID- 12908562 TI - Development of the epidermal growth factor receptor inhibitor Tarceva (OSI-774). AB - The epidermal growth factor receptor (EGFR) is a transmembrane receptor involved in the regulation of a complex array of essential biological processes such as cell proliferation and survival. Dysregulation of EGFR signaling network has been frequently reported in multiple human cancers and has been associated with the processes of tumor development, growth, proliferation, metastasis and angiogenesis. Inhibition of the EGFR was associated with antitumor effects in preclinical models. On the bases of these data, therapeutics targeting the EGFR were explore in clinical trials. Tarceva (OSI-774, OSI Pharmaceuticals, Uniondale, NY) is a small molecule selective inhibitor of the EGFR tyrosine kinase (TK). In preclinical studies, Tarceva inhibited the phosphorylation of the EGFR in a dose and concentration dependent manner resulting in cell cycle arrest and induction of apoptosis. In in vivo studies, the agent caused tumor growth inhibition and shoved synergistic effects when combined with conventional chemotherapy. Subsequent single agent phase I studies and phase I studies in combination with chemotherapy demonstrated that the agent has a good safety profile and induced tumor growth inhibition in a substantial number of patients with a variety of different solid tumor. Preliminary report from phase II studies confirmed the excellent tolerability of Tarceva as well as showed encouraging preliminary activity. Phase III studies have either been completed or are ongoing in several tumor types such as lung cancer and pancreatic cancer. In summary, Tarceva is a novel inhibitor of the EGFR TK which has shown promising activity in initial studies and is currently undergoing full development as an anticancer drug. PMID- 12908563 TI - Gefitinib (Iressa, ZD 1839) for non-small cell lung cancer (NSCLC): recents results and further strategies. AB - Phase I and II clinical studies demonstrated that gefinitib single agent therapy is well tolerated and provides clinically significant antitumor activity in patients with advanced NSCLC who have previously received prior treatment with cytotoxic chemotherapy. Large-scale studies to assess the clinical benefit of gefitinib versus placebo when combined with cisplatin/gemcitabine and carboplatin/paclitaxel as first-line treatment, however, did not prove that the three drug combinations be able to improve survival and compared to "standard" doubled chemotherapy. Therefore it can be summarized that gefinitib may provide evaluable addition to the therapeutic options available for the treatment of advanced NSCLC and may also have potential for use in early NSCLC. PMID- 12908564 TI - Mechanism of action of anti-HER2 monoclonal antibodies: scientific update on trastuzumab and 2C4. AB - The HER family of transmembrane tyrosine kinase receptors is composed of four members, BER1 to HER4. HER2 is a ligand-orphan receptor expressed in many human tumors and overexpressed in 25-30% of breast cancers. HER2 amplifies the signal provided by other receptors of the HER family by forming heterodimers. The essential role of HER2 in the HER signaling network led to the development of anti-HER2 monoclonal antibodies (MAbs) for cancer therapy. In particular, the humanized MAb trastuzumab (Herceptin) has antitumor activity against HER2 overexpressing human breast tumor cells and is widely used for the treatment of women with HER2 overexpressing breast cancers. Trastuzumab induces HER2 receptor downmodulation and, as a result, inhibits critical signalling pathways (i.e. ras Raf-MAPK and PI3K/Akt) and blocks cell cycle progression by inducing the formation of p27/Cdk2 complexes. Trastuzumab also inhibits HER2 cleavage, preceding antibody-induced receptor downmodulation, and this effect might contribute to its antitumor activity in some cancers. In vivo, trastuzumab inhibits angiogenesis and induces antibody-dependent cellular cytotoxicity. A limitation of trastuzumab is that its activity is largely restricted to breast cancers with the highest level of HER2 overexpression or HER2 gene amplification. However, there is a large population of breast cancers and of many other tumors that have low or moderate HER2 expression. In such tumors, HER2 functions as a preferred coreceptor to form heterodimers with HER1 (EGFR), HER3 or HER4. For this reason, a humanized monoclonal antibody, called 2C4, that targets the role of HER2 as a coreceptor is under active development. 2C4 binds to a different epitope of HER2 ectodomain than trastuzumab and sterically hinders HER2 recruitment in heterodimers with other HER receptors. This results in the inhibition of signalling by HER2-based heterodimers both in cells with low and high HER2 expression. In vitro and in vivo antitumor activity has been reported in a range of breast and prostate tumor models. Therefore, 2C4 may have potential against a wide variety of solid tumors. Phase I trials are underway. PMID- 12908565 TI - EORTC research and development: achievements and future perspectives. PMID- 12908566 TI - Renal tubular dysgenesis. PMID- 12908567 TI - Effect of corticosteroid and cyclophosphamide in IgA nephropathy patients with heavy proteinuria and/or moderate-severe pathological changes. AB - BACKGROUND: The treatment of immunoglobulin (Ig) A nephropathy remains a subject of controversy. To clarify if it is justified to use corticosteroid and cyclophosphamide in the treatment of IgA nephropathy patients with heavy proteinuria and/or a moderate-severe pathological grading of at least grade II, we retrospectively analyzed the medical data of IgA nephropathy patients in our hospital. METHODS: Totally 60 patients from 370 cases of biopsy-proved IgA nephropathy in our hospital between 1983 and 1999 met the selection criteria and were enrolled into this analysis. These patients received either corticosteroid alone or in combination with cyclophosphamide. The analysis of the progression of renal disease by serum creatinine and changes in the level of proteinuria were based on an on-treatment approach. Kidney survival curve was by Kaplan-Meier analysis and compared among different subgroups. RESULTS: Of the 60 patients, 20 cases (33.3%) (Group 1) reached a complete remission of proteinuria by prednisolone alone. The other cases received a combination therapy with corticosteroid and cyclophosphamide, which resulted in complete remission in 17 cases (28.3%) (Group 2), partial remission in 9 cases (15.0%) (Group 3) and complete resistance in 14 patients (23.3%) (Group 4). The renal survival rate was significantly worse in Group 4 (p < 0.05) as compared with other groups. CONCLUSIONS: We conclude that corticosteroid and combination therapy with corticosteroid and cyclophosphamide was effective treatment in 61.6% (Groups 1 and 2) of IgA nephropathy patients with heavy proteinuria and/or moderate-severe pathological lesions provided their initial serum creatinine level was < or = 3 mg/dl. PMID- 12908568 TI - Upper gastrointestinal endoscopy in children with upper gastrointestinal bleeding. AB - BACKGROUND: The purpose of this study was to evaluate the clinical presentations and features of upper gastrointestinal (UGI) endoscopy and the role of Helicobacter pylori in children with UGI bleeding. METHODS: A retrospective review was done of the medical records of 112 infants and children who presented with UGI bleeding. They underwent UGI endoscopy in the Taipei Veterans General Hospital, Taiwan between July 1996 and December 2000. Patients with hematemesis caused by swallowing caustic agents or foreign bodies were excluded. RESULTS: There were 112 patients involved in this study. Clinical manifestations included hematemesis (77 cases, 68.8%), tarry stool (37 cases, 33%) and hematochezia (9 cases, 8%). Fifty-three patients (47.3%) had underlying diseases, including neuromuscular deficits (17), hepatobiliary disease (17), renal insufficiency (8), Henoch-Schonlein purpura (HSP) (3), and cow milk allergy (3). Another 34 patients had a preceding acute febrile illness history. The endoscopic appearances included esophageal varices (10.7%), esophagitis or esophageal ulcers (30.4%), superficial gastric mucosal lesions (SGML) (44.6%), gastric ulcers (9.8%), duodenal ulcers (DU) (15.2%), and hemorrhagic erosive duodenitis (2.7%). Eleven cases (9.8%) showed negative findings. There were 13 (11.6%) patients receiving endoscopic hemostatic therapy. The rate of positive results in rapid urease tests in the patients without underlying diseases was 54.8%, which was significantly higher than that of the patients with underlying diseases (17.1%), while Helicobacter pylori was closely related to primary DU. CONCLUSIONS: In children with UGI bleeding, UGI endoscopy provides an accurate diagnostic tool and also provides therapeutic intervention when needed. SGML was the most common endoscopic finding. About 30% of the infants and children with UGI bleeding had history of acute febrile illness. Half of the children had underlying diseases. Helicobacter pylori plays a role in a subgroup of children with UGI bleeding, particularly in children with primary DU. PMID- 12908569 TI - The efficacy and safety of brimonidine 0.2% compared with timolol 0.5% in glaucoma: a randomized clinical trial on Taiwanese patients. AB - BACKGROUND: Brimonidine is a highly selective alpha-2 adrenergic receptor agonist with intraocular pressure (IOP) reducing effect. We conducted this study in Taiwan to compare the safety and efficacy ofbrimonidine 0.2% with timolol 0.5% for the treatment of glaucoma. METHODS: A prospective, randomized, single-masked, 1-month clinical efficacy and safety trial was conducted from March to September 2000. Forty glaucoma patients were enrolled--29 in the brimonidine group and 11 in the timolol group. Patients instilled their study medications twice daily for 4 weeks, and were followed at baseline visit, weeks 2 and 4. Demographic data, reduction of IOP, safety and adverse events were obtained and analyzed. RESULTS: Both drugs showed sustained ocular hypotensive efficacy in the study period. At baseline, the mean IOP was 24.48 +/- 2.29 mmHg in the brimonidine group and 23.32 +/- 0.82 mmHg in the timolol group. The IOP readings after treatment were significantly lower than their baseline levels in both groups at all visits (p < 0.001). At peak, the mean decreases from the baseline IOP ranged from 5.22 +/- 0.30 mmHg to 6.96 +/- 0.33 mmHg for brimonidine and from 4.55 +/- 0.49 mmHg to 6.64 +/- 0.53 mmHg for timolol. At trough, the mean decreases from baseline ranged from 3.72 +/- 0.32 mmHg to 4.55 +/- 0.32 mmHg for brimonidine and 3.82 +/- 0.52 mmHg to 4.27 +/- 0.51 mmHg for timolol. No significant between-group differences were seen at peak or trough at all visits. The clinical success rate was 86.2% in the brimonidine group and 81.8% in the timolol group, making no statistically significant difference between them (p = 0.817). 17.2% of patients in brimonidine group and 9.0% patients in timolol group reported mild adverse events. Ocular allergy occurred in 10.3% of patients in brimonidine group. No significant changes in visual acuity, biomicroscopy or ophthalmoscopy were observed in both groups. Mean systolic and diastolic blood pressure remained relatively stable in both groups except in week 2 (p = 0.016) when brimonidine had lower systolic blood pressure. However, brimonidine showed no significant difference in week 4 from baseline. The mean heart rate in the brimonidine group was relatively unchanged over the study period. Patients receiving timolol experienced statistically significant mean heart rate decreases from baseline (p = 0.020) in week 4. CONCLUSIONS: Topically applied twice daily for one month, brimonidine tartrate 0.2% has clinical effectiveness equivalent to timolol 0.5% in Taiwanese patients with glaucoma. It has a safe systemic profile with minimum effect on the heart. PMID- 12908570 TI - A potential alternative treatment of uncomplicated painful diverticular disease by trans-colonoscopic irrigation technique: a preliminary report. AB - BACKGROUND: Colonic diverticular disease is a common disorder in elder patients. Medical treatment was usually recommended as the first line management for this disease. However, the recurrence rate of such disorder is still high. In patients with severe complications such as abscess or fulminant inflammation, non-invasive diagnostic examination, abdominal CT scan for example, is recommended. Its most common symptom is repeated abdominal pain with disturbance of bowel habit. Many patients are found to be with diverticular disease only after colonoscopic examination. The aim of this study is to introduce a new irrigation-draining method and to evaluate its efficacy in treatment of uncomplicated painful colonic diverticular disease. METHODS: To reduce the risk of recurrence of acute diverticulitis and other severe complications, we introduce a transcolonoscopic irrigation technique for patients of uncomplicated diverticular disease by flushing out the obstructed fecalith from the diverticular sac in order to improve the drainage from the obstructed diverticular sac. RESULTS: Thirty-two patients of uncomplicated painful diverticular disease with obstructed fecalith impacted were treated by this technique. Clinical symptom improved in all of them and no complications developed during the mean follow-up period of 46 months. CONCLUSION: The results of this preliminary study suggest that this technique accomplished in the colonoscopic examination without additional therapeutic procedures. It provides another potential alternative to the conventional medical treatment for patients with uncomplicated diverticular disease. PMID- 12908571 TI - Surgical results of squamous cell carcinoma of the esophagus in young patients. AB - BACKGROUND: Squamous cell carcinoma of the esophagus (ESCC) is one of the ten most common cancers in Taiwan. Surgical resection is the mainstay of the treatment. Young patients with ESCC are reported to have poor surgical outcome. This retrospective and comparative study is to evaluate the clinicopathologic difference and survival between young and elderly patients. METHODS: From January 1985 to December 2000, totally 797 patients with esophageal cancer received esophagectomy with reconstruction at our institution. Among them, 12 patients who received neoadjuvant concurrent chemo-radiotherapy followed by esophagectomy were excluded from this study. The other 785 patients were divided into two groups. Group I included 67 patients aged 50 or younger (8.5%) and group II included 718 patients aged over 50 years (91.5%). The clinical data were collected by chart review and follow-up data were collected through outpatient department record or telephone contact. These two groups were analyzed and compared regarding sex ratio, exposure to tobacco or alcohol, surgical mortality rate and 5-year survival rate. RESULTS: There was no difference in gender distribution between young and old patients groups. More patients in the young patients group had the habits of alcohol and tobacco consumption than in the elder patients group. The surgical (30-day) mortality was 2.98% in the young patient group, and 5.43% in the elder group. The 5-year survival rate was better in young patients group than in elder patients group but there was no statistical significance. Survival in young patients group was significantly related to TNM staging system. CONCLUSIONS: More young patients with ESCC had the habit of smoking and drinking. There was no statistical difference between the groups in clinical variables, surgical mortality and survival. The survival rate was significantly related to the TNM tumor stage. PMID- 12908572 TI - NMP22 in transitional cell carcinoma of the urinary bladder. AB - BACKGROUND: Urinary bladder transitional cell carcinoma (TCC) is a common cancer encountered in the human urinary system. The detection of new and recurrent TCC of the urinary bladder was performed using nuclear matrix (NMP22) kit and compare to the urine cytology. METHODS: Ninety-two patients were included and divided to 3 groups. The first group was composed of patients for detection of recurrence of TCC of the urinary bladder in follow-up check. The second group consisted of patients who had microscopic or gross hematuria. The third group was volunteers. A urine sample was obtained from every enrolled patient. Followed by cystourethroscopy, the urine cytoloy, urine analysis and NMP 22 were performed respectively. A receiver operating characteristics (ROC) curve was constructed to determine NMP22 cutoff value for optimal sensitivity and specificity to detect bladder tumor. The data was then analyzed with Student-t test. RESULTS: ROC curve showed an optimal value of greater than 10 units per mL in differentiating positive from negative results. The sensitivity of NMP22 was 91.7% (22/24) and the specificity was 72.1% (49/68) in detection of TCC. The sensitivity and specificity of cytology were 37.5% (9/24) and 97.1% (66/68), respectively. The positive predictive values of NMP22 and cytology were 53.7% and 81.8%, individually. The false-positive rate of NMP22 was 27.9%. More sensitivity was observed in the NMP22 test with regards to high-grade and advanced-stage tumors. CONCLUSIONS: NMP22 is a highly sensitive tumor marker. However, high false positive rate and unsatisfactory positive predictive value are the drawbacks of this new tumor marker assay. There is inadequate evidence to consider total replacement of cytology with NMP22. But combination of NMP22 and cytology could be the most complete way to avoid unnecessary cystoscopic surveillance. PMID- 12908573 TI - Renal tubular dysgenesis in siblings. AB - Renal tubular dysgenesis is a recently recognized autosomal recessive condition characterized by absence or poor development of proximal convoluted tubules, clinical presentation of oligohydromnios, Potter sequence, and neonatal respiratory failure. Only a few cases of renal tubular dysgenesis have been diagnosed. We report 2 cases from a single family. Histologically, the renal cortices showed crowding of the glomeruli and primitive tubules, most of which demonstrated positive cytoplasmic staining for epithelial membrane antigen and peanut lectin. Electron microscopy of the kidneys of the second baby revealed absence of differentiated characteristics of the proximal tubules. PMID- 12908574 TI - Malignant renal epithelioid angiomyolipoma with aggressive behavior and distant metastasis. AB - Angiomyolipoma (AML) is a common renal tumor and mostly benign in entity. Malignant AML is extremely rare and most of them are found to be epithelioid AML histopathologically. We report the imaging features of a malignant epithelioid AML in a 58-year-old patient with liver and nodal metastases, and review the literatures. We have observed that AML of epithelioid subtype with tumoral necrosis may suggest the malignant change. PMID- 12908575 TI - Extraskeletal mesenchymal chondrosarcoma. AB - Extraskeletal mesenchymal chondrosarcoma is a rare malignancy. It is characteriged by a bimorphic histologic pattern, with a mesenchymal tissue mixed with malignant hyaline cartilage. It also has high incidence of local recurrence and distant metastasis. All cases have been reported fatal associated with this tumor in spite of complete surgical excision. A 35-year-old female presented with a palpable mass about 5 x 3 x 2.5 cm3 in size over her left forearm and was reported to be an extraskeletal mesenchymal chondrosarcoma. Marginal resection followed by regional radiotherapy was done. No local recurrence or distant metastasis was found two years after surgery. Thorough physical examination, series radiographic studies, and multiple sites of tumor biopsies before radical resection of the tumor may decrease the misdiagnostic rate for extraskeletal mesenchymal chondrosarcoma. Appropriate tumor treatment, close patient follow-up and timely treatment for local recurrence or distant metastasis may increase the survival rate. PMID- 12908577 TI - A voice from the past: where are the cats? PMID- 12908576 TI - Severely sustained vomiting as the main symptom in a man with thyrotoxicosis. AB - Thyrotoxicosis has a variety of presentations. Vomiting as a main presenting symptom of thyrotoxicosis is uncommon. We report a 40-year-old male with thyrotoxicosis who presented with sustained vomiting as the main symptom. He also had weight loss, about 10 kg over this 20-day period, and dizziness, particularly in the upright position. Esophagogastroduodenoscopy and abdominal ultrasonography were negative. Laboratory data were unremarkable except serum T4 of 21.2 microg/dl, T3 of 574 ng/dl and TSH < 0.03 microIU/ml. The patient's serum microsomal antibody was positive at a titer of 1:409,600, but serum thyroglogulin antibody was negative at a titer of less than 1:100. The symptoms improved after adminstroction of propylthyrouracil and propranol. A total of 31 such cases have been reported in English literature. The mean age of the patients was 46 +/- 14 year with a range of 19 to 68 years. Only 4 patients, including ours, were male. Weight loss was found in about half of them and might be an important clue. Thyrotoxicosis should be considered in differential diagnosis of unexplained vomiting. PMID- 12908578 TI - Inhibitory mechanism of taurine on the platelet-derived growth factor BB-mediated proliferation in aortic vascular smooth muscle cells. PMID- 12908579 TI - Taurine on sino-atrial nodal cells: Ca2+-dependent modulation. PMID- 12908580 TI - Cellular characterization of taurine transporter in cultured cardiac myocytes and nonmyocytes. PMID- 12908581 TI - Taurine transporter in cultured neonatal cardiomyocytes: a response to cardiac hypertrophy. PMID- 12908582 TI - Effect of taurine and coenzyme Q10 in patients with acute myocardial infarction. PMID- 12908583 TI - Taurine reduces renal ischemia/reperfusion injury in the rat. PMID- 12908584 TI - Taurine-depleted heart and afterload pressure. PMID- 12908585 TI - Taurine reduces mortality in diabetic rats: taurine and experimental diabetes mellitus. PMID- 12908586 TI - The effect of dietary taurine supplementation on plasma and urinary free amino acid concentrations in diabetic rats. PMID- 12908587 TI - Insulin-stimulated taurine uptake in the rat retina. PMID- 12908588 TI - The effect of taurine supplementation on patients with type 2 diabetes mellitus. PMID- 12908589 TI - Hypoglycemic effects of taurine in the alloxan-treated rabbit, a model for type 1 diabetes. PMID- 12908590 TI - Taurine and cellular volume regulation in the hippocampus. PMID- 12908591 TI - Regulation of the volume-sensitive taurine efflux pathway in NIH3T3 mouse fibroblasts. PMID- 12908592 TI - Taurine counteracts cell swelling in rat cerebrocortical slices exposed to ammonia in vitro and in vivo. PMID- 12908593 TI - Cloning of human intestinal taurine transporter and production of polyclonal antibody. PMID- 12908594 TI - Transactivation of TauT by p53 in MCF-7 cells: the role of estrogen receptors. PMID- 12908595 TI - Gating of taurine transport: role of the fourth segment of the taurine transporter. PMID- 12908596 TI - Finding of TRE (TPA responsive element) in the sequence of human taurine transporter promoter. AB - Activity of the taurine transporter (TAUT) is regulated by signal transduction in response to diverse stimuli including tumor promoters such as phobol ester. Regulation of the transcription rate of TAUT appears to play an important role in exerting biological roles of taurine in mammalian tissues in adverse environments. Although cDNA of human TAUT has been cloned and sequenced in placenta, thyroid cells, and retinal pigment epithelial cells, the promoter region of TAUT has never been reported. In order to clone the upstream region of the human TAUT promoter, we have compared TAUT cDNA sequences with the entire human genome sequence. Polymerase chain reaction (PCR) was performed from genomic DNA prepared from a SK-Hep-1 cell line for the amplification of the TAUT promoter region including the partial exon (150 bp) and the 5' untranslated region (UTR, 380 bp). The PCR product of the promoter region, which was 1800 bp long, was ligated into the pGEM-T vector, and sequenced. The 5' flanking region of the TAUT promoter was analysed for the identification of enhancer and regulation motifs. Surprisingly we found the consensus TPA responsive element (TGAGTCAG) which is responsible for gene regulation by the protein kinase C (PKC)-mediated signal transduction pathway. The well known fact that proto-oncogene AP1 (cFos/cJun heterodimer or cJun/cJun homodimer) binds to TRE implies that TAUT expression might be closely linked to tumor promotion. Since AP1 activity is also tightly regulated in nerve cells, AP1-regulated TAUT transcription might be an important step in nerve cell function. Furthermore, the TFIID binding site, cap signal for transcription initiation, PEA3 motif, heat shock factor binding motif, and many other motifs were found in the TAUT promoter region, and require characterization. PMID- 12908597 TI - Protein kinase C and cAMP mediated regulation of taurine transport in human colon carcinoma cell lines (HT-29 & Caco-2). PMID- 12908599 TI - Isovolumetric regulation in mammal cells: role of taurine. PMID- 12908598 TI - Characterization of transcriptional activity of taurine transporter using luciferase reporter constructs. AB - Although taurine transporter (TAUT) activity has been known to be regulated by diverse intracellular and extracellular factors involved in the signal transduction pathway, such as protein kinase C, intracellular Ca concentration, and glucocorticoids, little is known concerning the underlying mechanisms. Evidence suggests that such stimulation-mediated changes in TAUT activity in mammalian cells are partly achieved through the modulation of TAUT transcription activity. In order to better understand the regulation of TAUT transcription activity and subsequently the role of taurine in the signal transduction pathway, we have cloned and sequenced the 5' flanking region of the human TAUT gene, and characterized the TAUT promoter region in human cells. For these reasons, the TAUT luciferase reporter vector was constructed using the 5' flanking region of the TAUT gene (1800 bp). The TAUT luciferase reporter vector was then transfected into SiHa cells, and luciferase activity was measured. The construct containing its own promoter of TAUT (pGL3 b TAU31) showed a 10 fold higher luciferase activity compared to the value found in the empty vector (pGL3 b). This implies a functional transcription of the homologous TAUT promoter. Similar results were obtained with the exon deleted construct [pGL3 b TAU31(-e)]. We also constructed the TAUT luciferase reporter gene (pGL3 pro TAU13) using a heterologous promoter. About 2.5 fold higher luciferase activity was observed in cells transfected with this construct containing the heterologous promoter compared to the value found in the control vector (PGL3 pro). PMID- 12908600 TI - Osmosensitive taurine release: does taurine share the same efflux pathway with chloride and other amino acid osmolytes? PMID- 12908601 TI - Electrophysiological properties of the mouse Na+/Cl(-)-dependent taurine transporter (mTauT-1): steady-state kinetics: stoichiometry of taurine transport. PMID- 12908602 TI - Taurine uptake and release by the pancreatic beta-cells: taurine transport in beta-cells. PMID- 12908603 TI - Regulation of intestinal taurine transporter by cytokines. PMID- 12908604 TI - Determination of taurine and hypotaurine in animal tissues by reversed-phase high performance liquid chromatography after derivatization with dabsyl chloride. PMID- 12908605 TI - Immunohistochemical localiztion of taurine in the rat stomach. PMID- 12908606 TI - Modulation of taurine on CYP3A4 induction by rifampicin in a HepG2 cell line. PMID- 12908607 TI - Effect of acute ethanol administration on S-amino acid metabolism: increased utilization of cysteine for synthesis of taurine rather than glutathione. AB - Alterations in the hepatic metabolism of S-amino acids were examined in male rats injected with a single dose of ethanol (3 g/kg, i.p.). The hepatic concentrations of methionine and S-adenosylhomocysteine (SAH) were increased, but S adenosylmethionine (SAM), cysteine, and glutathione (GSH) decreased rapidly following ethanol administration. The activities of methionine adenosyltransferase (MAT), cystathionine beta-synthase (CbetaS) and cystathionine gamma-lyase (CgammaL) were all inhibited. Gamma-glutamylcysteine synthetase (GCS) activity was increased from t = 8 hr, but hepatic glutathione (GSH) level did not return to control for 48 hr. Both hepatic hypotaurine and taurine levels were increased immediately, which were reduced to below control from t = 18 hr. Changes in the serum concentration of taurine were consistent with results observed in the liver. Cysteine dioxygenase (CDO) activity was increased rapidly, but declined from t = 24 hr. The results indicate that an acute dose of ethanol induces significant alterations in the metabolism of S-amino acids in the liver. Ethanol depresses the cysteine availability for GSH synthesis not only by inhibiting the transsulfuration reactions but also by enhancing its irreversible catabolism to taurine via hypotaurine. The physiological significance of this finding is discussed. PMID- 12908608 TI - Effects of dietary taurine supplementation on hepatic morphological changes of rats in diethylnitrosamine-induced hepatocarcinogenesis. PMID- 12908609 TI - Effect of taurine on cholesterol degradation and bile acid pool in rats fed a high-cholesterol diet. PMID- 12908610 TI - Effect of alpha-tocopherol and taurine supplementation on oxidized LDL levels of middle aged Korean women during aerobic exercise. PMID- 12908611 TI - Dietary taurine intake and serum taurine levels of women on Jeju Island. AB - The purpose of this study was to investigate the dietary taurine intake and serum taurine levels of women on Jeju Island in Korea. Sixty six married women aged 43.5 +/- 7.1 volunteered for this study: 34 from the city area and 32 from two fishing-farming areas. Diet samples were collected from the participants; the samples included three meals (breakfast, lunch and supper), including snacks, drinks and whatever else the participants had eaten for 24 hours. Taurine levels in the diet and serum were determined as the dabsyl derivative by HPLC with a Rf detector. The intake of taurine ranged from 8.4 to 767.6 mg/day and its mean value was 163.9 +/- 150.2 mg/day (mean +/- SD). There was a significant difference between the two groups: 114.9 +/- 78.7 for the women from the city area and 215.9 +/- 187.9 mg/day for the women from the fishing-farming areas (p<0.001). The taurine intake of the total diet, including all snacks and drinks, was 2300 +/- 584 g/day for the city area and 2342 +/- 528 g/day for the fishing farming areas. The daily protein intake was 58.8 +/- 16.4 g for the women of the city area and 65.5 +/- 17.1 g for the women of the fishing-farming areas. There was a significant correlation between the intake of fish/shellfish and taurine (p=0.001) while there was no correlation between the intake of protein and taurine (p=0.057). The taurine levels in serum ranged from 68.6 to 261.6 micromol/L and the mean value was 169.7 +/- 41.5 micromol/L. There was no significant difference between the women from the city area and the women from the fishing-farming areas in serum taurine levels. The correlations of serum taurine levels with serum retinol levels (p=0.016) and alpha-tocopherol (p=0.014) levels were significant. These results suggest that taurine intake is dependent on the fish/shellfish intake and that taurine may play an important role in the retention of antioxidative nutrients. PMID- 12908612 TI - Effect of the obesity index on plasma taurine levels in Korean female adolescents. PMID- 12908613 TI - Regional differences in the dietary taurine intake in Korean college students. PMID- 12908614 TI - Taurine concentration in human blood peripheral lymphocytes: major depression and treatment with the antidepressant mirtazapine. AB - Major depression is a serious disease with various systemic effects, including dysfunction of the immune response. Taurine has been known to be related to certain modifications of the immune system. The aim of this study was to determine the taurine concentration in lymphocytes of patients with major depression and to evaluate the influence of the antidepressant treatment with mirtazapine for six weeks on the levels of taurine. Gamma-aminobutyric acid, aspartate, glutamate and glutamine were also determined. Taurine, aspartate and glutamine levels were increased in the lymphocytes of depressed patients before mirtazapine treatment compared to the control group, and were normalized after treatment. Gamma-aminobutyric acid and glutamate did not differ between patients and controls. There was a significant and positive correlation between the severity of the disorder, measured by the Hamilton Rating Scale, and the concentration of taurine in the lymphocytes of depressed patients before treatment. This correlation was not observed after treatment and neither was there a correlation observed for the other amino acids. The present observations could be an indication of the relevance of taurine as a protective agent in the lymphocytes of patients with severe depression, and could be the result of modifications of taurine transport or efflux processes. PMID- 12908615 TI - Why is taurine cytoprotective? PMID- 12908616 TI - Taurine, analogues and bone: a growing relationship. PMID- 12908617 TI - Anti-inflammatory activities of taurine chloramine: implication for immunoregulation and pathogenesis of rheumatoid arthritis. PMID- 12908619 TI - Taurine chloramine inhibits production of inflammatory mediators and iNOS gene expression in alveolar macrophages; a tale of two pathways: part II, IFN-gamma signaling through JAK/Stat. PMID- 12908618 TI - Taurine chloramine inhibits production of inflammatory mediators and iNOS gene expression in alveolar macrophages; a tale of two pathways: part I, NF-kappaB signaling. PMID- 12908620 TI - Production of nitric oxide by activated microglial cells is inhibited by taurine chloramine. PMID- 12908621 TI - Production of inflammatory mediators by activated C6 cells is attenuated by taurine chloramine inhibition of NF-kappaB activation. PMID- 12908622 TI - Taurine is involved in oxidation of IkappaB alpha at Met45: N-halogenated taurine and anti-inflammatory action. PMID- 12908623 TI - The combined treatment with taurine and niacin blocks the bleomycin-induced activation of nuclear factor-kappaB and lung fibrosis. PMID- 12908624 TI - Taurine reduces lung inflammation and fibrosis caused by bleomycin. PMID- 12908625 TI - Taurine and the lung: which role in asthma? PMID- 12908626 TI - Effect of taurine and other antioxidants on the growth of colon carcinoma cells in the presence of doxorubicin or vinblastine in hypoxic or in ambient oxygen conditions: effect of antioxidants on the action of antineoplastic drugs in MDR and non-MDR cells. PMID- 12908627 TI - The role of taurine in cerebral ischemia: studies in transient forebrain ischemia and embolic focal ischemia in rodents. AB - Sudden cessation of blood flow to the brain results in a series of events that either result in rapid loss of brain cells or delayed neuronal injury in certain vulnerable regions of the brain. Research over the last three decades has allowed for a better understanding of how neurons and other brain cells die from the effects of ischemia and hypoxia in the central nervous system. Excitatory and inhibitory neurotransmitters exist in a very precise balance for normal function of the brain. Ischemia very rapidly disrupts this balance resulting in a rapid build-up of excitatory neurotransmitters, especially glutamate in the extracellular space. The increased glutamate together with energy loss opens a number of different types of calcium and sodium channels resulting in the build up of these ions in neurons, leading to cellular dysfunction and death. While most ischemia research has focused on antagonism of excitatory amino acids, there are some reports on enhancement and amplification of inhibitory responses in focal and global ischemia. The majority of work relates to potentiation of GABA, either endogenous or through GABA potentiating medications. Taurine has neuroinhibitory properties and may also have potential for neuroprotection in cerebral ischemia. This present review focuses on the role of taurine as a neuroprotective agent, possibly acting through several different inhibitory mechanisms. Taurine may inhibit neurotransmitter release and may result in normal intracellular osmolality. In transient global ischemia in gerbils, we studied in vivo microdialysis of amino acids before, during and after ischemia. We were able to show that taurine resulted in attenuation of glutamate during ischemia (however did not reach significance). In similar experiments, neuronal damage was assessed in the hippocampus. Our results show 48% damage in taurine treated animals, 60% in alanine treated animals and 69% in control groups (trend towards protection but again did not reach significance) Focal ischemia was induced by embolizing a thrombus into the distal internal carotid artery and origin of the middle cerebral artery. Again, in studies where we compared taurine to a placebo treated animal, there was no significant decrease in the amount of damage with taurine. There are reports in the literature that taurine may attenuate neuronal injury during ischemia. Our studies in two models of cerebral ischemia in rodents did not reveal neuronal protection. It is possible that higher doses or possibly prolonged use of taurine may show better results. Taurine may also potentially offer additive protective effects when used in combination with thrombolysis or other neuroprotective agents. Further studies are necessary to better understand the potential for taurine as a neuroprotective agent in cerebral ischemia. PMID- 12908628 TI - Studies on taurine efflux from the rat cerebral cortex during exposure to hyposmotic, high K+ and ouabain-containing aCSF. PMID- 12908629 TI - Interactions of taurine and adenosine in the mouse hippocampus in normoxia and ischemia. PMID- 12908630 TI - Involvement of nitric oxide in ischemia-evoked taurine release in the mouse hippocampus. PMID- 12908631 TI - Effect of ammonia on taurine transport in C6 glioma cells. AB - Both short-term and prolonged treatment with ammonia stimulate Tau efflux from C6 glioma cells, which confirms earlier observations with different CNS preparations. In addition, prolonged, but not short-term treatment reversibly decreases Tau uptake. The results are consistent with in vivo observations showing robust extracellular accumulation of Tau in the CNS under hyperammonemic conditions. Enhancement of Tau efflux by short term ammonia treatment is associated with its passage via a channel (or channels) which in contrast to the release from control cells is (are) not inhibited by NA. However, the increased Tau efflux does not appear to involve active Tau transport. Insensitivity to NA distinguishes ammonia-dependent Tau efflux from C6 cells from that reported in other CNS-derived preparations. Tau release evoked by prolonged ammonia treatment likewise shows a NA-insensitive component, but also a component associated with activation of Tau transport in a reverse mode. Increased outward transport of Tau may be associated with transiently increased expression of TauT mRNA. Mutual relation of the two components, but also the identity of the NA-insensitive efflux route with the anion channels so far described remains to be analyzed. PMID- 12908632 TI - Taurine and hypotaurine dynamics in activated C6 glioma: the effects of lipopolysaccharide (LPS) and taurine administration on intracellular hypotaurine and taurine dynamics in C6 glioma. PMID- 12908633 TI - The anti-craving taurine derivative acamprosate: failure to extinguish morphine conditioned place preference. PMID- 12908634 TI - Ethanol-induced taurine efflux: low dose effects and high temporal resolution. PMID- 12908635 TI - Tyrosine kinases and taurine release: signaling events and amino acid release under hyposmotic and ischemic conditions in the chicken retina. PMID- 12908636 TI - Effect of taurine on regulation of GABA and acetylcholine biosynthesis. PMID- 12908637 TI - Taurine effect on neuritic outgrowth from goldfish retinal explants in the absence and presence of fetal calf serum. AB - Post-crush goldfish retinal explants were cultured in the absence of fetal calf serum and in the presence of calcium, albumin, glucose or taurine. There was an increase in length and density of the neurites in a dose-dependent manner when fetal calf serum was added to the medium. The addition of calcium to the basic medium, Leibovitz, produced a small increase in neuritic outgrowth at low concentrations, but inhibited outgrowth at higher levels. Albumin did not significantly modify neuritic outgrowth. Glucose, at variable concentrations, produced a bell-shaped increase in length and density of the neurites. Taurine, in the absence of fetal calf serum, produced a small increase in neuritic outgrowth at 10 mM. In the presence of glucose, taurine stimulated outgrowth was less efficient than in the presence of fetal calf serum. In order to investigate the mechanisms of action of taurine as a trophic agent, the culture medium in the absence of fetal calf serum will be supplemented with glucose. PMID- 12908638 TI - Prevention of epileptic seizures by taurine. AB - Parenteral injection of kainic acid (KA), a glutamate receptor agonist, causes severe and stereotyped behavioral convulsions in mice and is used as a rodent model for human temporal lobe epilepsy. The goal of this study is to examine the potential anti-convulsive effects of the neuro-active amino acid taurine, in the mouse model of KA-induced limbic seizures. We found that taurine (43 mg/Kg, s.c.) had a significant antiepileptic effect when injected 10 min prior to KA. Acute injection of taurine increased the onset latency and reduced the occurrence of tonic seizures. Taurine also reduced the duration of tonic-clonic convulsions and mortality rate following KA-induced seizures. Furthermore, taurine significantly reduced neuronal cell death in the CA3 region of the hippocampus, the most susceptible region to KA in the limbic system. On the other hand, supplementation of taurine in drinking water (0.05%) for 4 continuous weeks failed to decrease the number or latency of partial or tonic-clonic seizures. To the contrary, we found that taurine-fed mice showed increased susceptibility to KA-induced seizures, as demonstrated by a decreased latency for clonic seizures, an increased incidence and duration of tonic-clonic seizures, increased neuronal death in the CA3 region of the hippocampus and a higher post-seizure mortality of the animals. We suggest that the reduced susceptibility to KA-induced seizures in taurine-injected mice is due to an increase in GABA receptor function in the brain which increases the inhibitory drive within the limbic system. This is supported by our in vitro data obtained in primary neuronal cultures showing that taurine acts as a low affinity agonist for GABA(A) receptors, protects neurons against kainate excitotoxic insults and modulates calcium homeostasis. Therefore, taurine is potentially capable of treating seizure-associated brain damage. PMID- 12908639 TI - Taurine regulates mitochondrial calcium homeostasis. AB - We have investigated the protective role of taurine in glutamate-mediated cell death and the involvement of mitochondria in this process. In cultured cerebellar granule cells, glutamate induces a rapid and sustained elevation in cytoplasmic free calcium ([Ca2+]i), causing the collapse of the mitochondrial electrochemical gradient (MtECG) and subsequent cell death. We found that pre-treatment with taurine, did not affect the level of calcium uptake with glutamate but rather reduced its duration; the calcium increase was transient and returned to basal levels about 10 min after adding glutamate. Furthermore, taurine reduced mitochondrial calcium concentration under non-depolarizing conditions. Treatment of cerebellar granule cells with taurine enhanced mitochondrial activity as measured by rhodamine uptake, both in the presence or absence of glutamate. We conclude that taurine prevents or reduces glutamate excitotoxicity through both the enhancement of mitochondrial function and the regulation of intracellular (cytoplasmic and mitochondrial) calcium homeostasis. PMID- 12908640 TI - Taurine in aging and models of neurodegeneration. PMID- 12908641 TI - Taurine stimulation of calcium uptake in the retina: mechanism of action. PMID- 12908642 TI - The nature of taurine binding in the retina. PMID- 12908643 TI - Predictors of retention in dual-focus self-help groups. AB - Attendance at 12-step groups has been found useful in maintaining abstinence from substance use; many members disengage early, missing out on potential benefits. New 12-step based groups have emerged to address the recovery needs of the many substance users with psychiatric comorbidity. Little is known about factors associated with retention in 12-step, especially in this population. This study sought to identify predictors of retention over a one-year period among members of a dual-focus 12-Step fellowship (N = 276). Using multivariate analysis, the following baseline characteristics were associated with greater retention one year later: older age, more lifetime arrests, abstinence in the pre-baseline year, more psychiatric symptoms in the pre-baseline year, not taking psychiatric medication, being more troubled by substance abuse than by mental health, and greater level of self-efficacy for recovery; residing in supported housing and being enrolled in outpatient treatment at follow-up were also significantly associated with better retention. Clinical implications to enhance retention in specialized 12-step groups are discussed. PMID- 12908644 TI - Relationship between self-report and clinician-rated impairment in depressed outpatients. AB - This study examined the potential of using the Global Assessment of Functioning (GAF) scale as a valid measure of patient-reported status and predictor of three month outcomes for 59 patients seeking outpatient treatment for depression. A moderate relationship between the GAF and patient report at baseline and follow up suggests it may be clinically useful for routine monitoring of depressed patients. PMID- 12908645 TI - Innovation and implementation in mental health services for homeless adults: a case study. AB - Seeking to identify conditions that support newly implemented evidence-based practices, this case study examined an implementation in which an existing agency was invited to move into the neighboring county to introduce its "housing first" practice with seriously mentally ill homeless adults. Using a constructivist methodology to elicit the narratives of key actors and observers about the implementation and its attendant controversy, this study found three issues at the core of actors' experiences: mode of presentation, use of an outside agency and the questioned uniqueness of the new practice. Barriers rather than facilitators dominated participants' interpretations of events despite significant researcher-observed facilitators. PMID- 12908646 TI - Literacy and comprehension of Beck Depression Inventory response alternatives. AB - This study explored the relationship of literacy level to comprehension of response alternatives from the Beck Depression Inventory (BDI). Twenty-nine adult basic education students, 14 senior citizens, and 20 oncology patients were given a brief literacy assessment and a comprehension test of selected BDI response alternatives. Three raters with clinical experience assessed comprehension. Lower literacy was significantly associated with less comprehension, even when other factors were controlled, yet comprehension problems were still found among higher literacy groups. The percent of participants who correctly paraphrased the selected BDI response alternatives ranged from 0 to 60%. These preliminary results suggest that poor comprehension of mental health measures is not only prevalent, but also cannot be estimated from patients' education or assessed reading levels. PMID- 12908647 TI - Improving the vocational status of patients with long-term mental illness: a randomised controlled trial of staff training. AB - OBJECTIVE: To investigate the impact of training CMHT members in the practice of Individual Placement and Support on the vocational status of long-term patients. METHOD: Six CMHTs received vocational training by a work co-ordinator; four continued with standard care. The best vocational status of the 1037 subjects was ascertained after one year. Factors associated with improvement in vocational status were identified. RESULTS: There was no difference in change of vocational status. Age, previous employment and diagnosis influenced outcome. CONCLUSION: Training in IPS at team level did not improve employment status. A dedicated, vocational worker appears to be essential for successful IPS. PMID- 12908648 TI - Job satisfaction among mental health professionals in Rome, Italy. AB - Although various surveys on job satisfaction have been performed in mental health care settings, no studies have investigated in-depth the level of satisfaction with the various aspects of work in Italian mental health services. In the present study, all clinical mental health staff working in a large psychiatric catchment area in Rome were invited to anonymously complete a previously validated questionnaire designed to measure job satisfaction among mental health professionals. Of the total 236 health professionals, 196 (83%) agreed to participate. Most participants were not completely satisfied with many aspects of their job, and many were not even moderately satisfied. The level of satisfaction increased with age, and it was significantly lower among hospital-ward staff compared to the staff of outpatient clinics or residential facilities, even after adjusting for age, gender, profession, work setting, and time in current job, using a multiple logistic regression model. Our findings suggest that interventions aimed at increasing job satisfaction among Italian mental health professionals might be warranted, particularly among hospital-ward staff. PMID- 12908649 TI - Drug information sources and antidepressant adherence. AB - The purpose of the study was to examine how patient sources of antidepressant information were related to antidepressant adherence. Eight community pharmacies in central North Carolina participated in the study. A research assistant interviewed eighty-one English-speaking patients who were picking up antidepressant refill prescriptions. Patients most frequently reported receiving antidepressant information from: pharmacists (58%), primary care physicians (50.6%), mental health specialists (40.7%), friends or family members (32.1%), and the Internet (18.5%). Patients experiencing more side effects were significantly less adherent to their antidepressants. Patients who received antidepressant information from more sources were significantly more adherent to their regimen than patients who received information from fewer sources. PMID- 12908650 TI - Use of the balanced scorecard in health care. AB - Since Kaplan and Norton published their article proposing a balanced scorecard, the concept has been widely adopted by industry and health care provider organizations. This article reviews the use of the balanced scorecard in health care and concludes that the balanced scorecard: (1) is relevant to health care, but modification to reflect industry and organizational realities is necessary; (2) is used by a wide range of health care organizations; (3) has been extended to applications beyond that of strategic management; (4) has been modified to include perspectives, such as quality of care, outcomes, and access; (5) increases the need for valid, comprehensive, and timely information; and (6) has been used by two large-scale efforts across many health care organizations in a health care sector, which differ, namely in the units of analysis, purposes, audiences, methods, data, and results. PMID- 12908651 TI - The hospital bond market and the AHERF bankruptcy. AB - An analysis of hospital, tax-exempt bonds issued before and after the Allegheny Health, Education, and Research Foundation (AHERF) bankruptcy demonstrated that despite the decline in market rates for tax-exempt securities in the post period, bonds issued by hospitals and systems carried higher coupon rates than they did in the pre period. There was a significant decline in the proportion of hospital/system bonds that were insured from the pre to the post period. Bond insurance firms tightened their credit criteria after the bankruptcy, which may explain, in part, why the proportion of insured bonds declined. We conclude that hospital bonds are now viewed as riskier instruments than they were prior to the AHERF bankruptcy. This is reflected in higher coupon rates for both insured and uninsured bonds and fewer insured bond issues. This decline in hospital creditworthiness comes at a time when many hospitals need to replace aging assets and acquire new technologies in response to increased inpatient utilization. PMID- 12908652 TI - Benchmarking variation in coding accuracy across the United States. AB - The objective of this study was to measure the consistency of coded medical data through information managers' reports of the overall coding error level in patients' medical records. Using a cross-sectional design, we examined the reported percent of records containing coding errors significant enough to change a diagnostic related group (DRG). Results indicate about 87 percent, 9 percent, and 5 percent of respondents reported that significant coding errors existed in less than 5 percent, 6-10 percent, and greater than 10 percent of the medical records in their institutions, respectively. Significant variation was found in the accuracy and consistency of coding practice and associated data quality across key demographic and organizational variables. Significantly large error rates in coded data exist in some organizations. Given variations across key demographic characteristics, providers may tend to distrust all coded data, when aggregated. As the United States moves toward an evidence-based medicine environment, the use of current patient data classification methods may be of limited value without increased attention to coding practices. PMID- 12908653 TI - Disparity in coding concordance: do physicians and coders agree? AB - Increasing demands for large-scale comparative analysis of health care costs has led to a similar demand for consistently classified data. Evidence-based medicine demands evidence that can be trusted. This study sought to assess managers' observed levels of agreement with physician code selections when classifying patient data. Using a non-sampled research design of both mailed and telephone surveys, we employ a nationwide cross-section of over 16,000 accredited US medical record managers. As a main outcome measure, we evaluate reported levels of agreement between physician and information manager code selections made when classifying patient data. Results indicate about 19 percent of respondents report that coder-physician classification disagreement occurred on more than 5 percent of all patient encounters. In some cases, disagreement occurred in 20 percent or more instances of code selection. This phenomenon shows significant variation across key demographic and market indicators. With the growing practice of measuring coded data quality as an outcome of health care financial performance, along with adoption of electronic classification and patient record systems, the accuracy of coded data is likely to remain uncertain in the absence of more consistent classification and coding practices. PMID- 12908654 TI - Nurse staffing, quality, and financial performance. AB - In examining the relationship among nurse staffing, quality of care, and financial performance, prior empirical studies used competing measures and applied different levels of analysis. Using longitudinal data from 1990 through 1995, our study applied a dynamic econometric model to evaluate whether hospitals that changed their nurse staffing and quality of care affected their financial performance. Sampling 422 hospitals over this study period, we found a statistically significant increase in operating costs when registered nurse levels increase, but no statistically significant decrease in profit. Higher levels of non-nurse staffing caused higher operating expenses, as well as lower profits. PMID- 12908655 TI - Health systems performance: a statewide analysis. AB - In this study, we examine the statewide consumption of health care resources. We assume that the physicians, hospital beds, nurses, and pharmaceuticals represent inputs to the health care delivery system required to treat the population of each state. Since health care resources needed by the various age groups of the population are different, we categorize the population into under 18 years of age, 18 to under 65 years of age, and 65 years of age and older. We develop efficient production frontiers and estimate the overall efficiency of individual states, and also the efficiencies of four inputs. In addition, we estimate the excess number of physicians, nurses, hospital beds, and pharmaceuticals consumed. This study should be useful for the states to help identify appropriate policy measures to make their health delivery systems more efficient. PMID- 12908656 TI - Guest editors' introduction: what can large pragmatic clinical trials do for public mental health care? PMID- 12908657 TI - Approaches to multisite clinical trials: the National Institute of Mental Health perspective. AB - The Division of Services and Intervention Research of the National Institute of Mental Health (NIMH) utilizes a variety of methodologies and approaches in the development of its clinical trials program. In this article, we describe the need for large multisite trials; the mechanisms for addressing this need; and the various approaches that have been used. In the course of carrying out this initiative, we have created opportunities for the Institute and its trial investigators to receive advice and input from the field. We describe the role and function of the trial coordinating center and NIMH staff. We identify the first steps to be taken in the initiation of a trial and highlight the opportunity for ancillary studies. Finally, we enumerate some of the pitfalls of large clinical trials and discuss measures taken to anticipate and address them. PMID- 12908658 TI - The National Institute of Mental Health Clinical Antipsychotic Trials of Intervention Effectiveness (CATIE) project: schizophrenia trial design and protocol development. AB - The National Institute of Mental Health initiated the Clinical Antipsychotic Trials of Intervention Effectiveness (CATIE) program to evaluate the effectiveness of antipsychotic drugs in typical settings and populations so that the study results will be maximally useful in routine clinical situations. The CATIE schizophrenia trial blends features of efficacy studies and large, simple trials to create a pragmatic trial that will provide extensive information about antipsychotic drug effectiveness over at least 18 months. The protocol allows for subjects who receive a study drug that is not effective to receive subsequent treatments within the context of the study. Medication dosages are adjusted within a defined range according to clinical judgment. The primary outcome is all cause treatment discontinuation because it represents an important clinical endpoint that reflects both clinician and patient judgments about efficacy and tolerability. Secondary outcomes include symptoms, side effects, neurocognitive functioning, and cost-effectiveness. Approximately 50 clinical sites across the United States are seeking to enroll a total of 1,500 persons with schizophrenia. Phase 1 is a double-blinded randomized clinical trial comparing treatment with the second generation antipsychotics olanzapine, quetiapine, risperidone, and ziprasidone to perphenazine, a midpotency first generation antipsychotic. If the initially assigned medication is not effective, subjects may choose one of the following phase 2 trials: (1) randomization to open-label clozapine or a double blinded second generation drug that was available but not assigned in phase 1; or (2) double-blinded randomization to ziprasidone or another second generation drug that was available but not assigned in phase 1. If the phase 2 study drug is discontinued, subjects may enter phase 3, in which clinicians help subjects select an open-label treatment based on individuals' experiences in phases 1 and 2. PMID- 12908659 TI - Assessing clinical and functional outcomes in the Clinical Antipsychotic Trials of Intervention Effectiveness (CATIE) schizophrenia trial. AB - Schizophrenia is a symptomatically heterogeneous disorder characterized by the presence of positive and negative symptoms, and variable impairment in community functioning. Given the diversity of symptom presentations and functioning associated with schizophrenia, one of the key challenges facing the Clinical Antipsychotic Trials of Intervention Effectiveness (CATIE) schizophrenia trial was the selection of efficient assessment measures appropriate to a community based effectiveness trial. This article describes the rationale for the measurement approach adopted for the trial, provides a brief overview of the selected measures, and describes the process of training assessment raters for a large and geographically dispersed study group. PMID- 12908661 TI - Clinical Antipsychotic Trials of Intervention Effectiveness (CATIE): Alzheimer's disease trial. AB - This article describes the development of the protocol for the Clinical Antipsychotic Trials of Intervention Effectiveness (CATIE) Alzheimer's disease trial, which was developed in collaboration with the National Institute of Mental Health to assess the effectiveness of atypical antipsychotics for psychosis and/or agitation occurring in outpatients with Alzheimer's disease. The article provides a detailed description of the methodology used in the trial as well as the clinical outcomes and effectiveness measures incorporated into it, discussing the most salient issues encountered in developing the design of the trial, as well as the unique features of the trial. PMID- 12908660 TI - Neurocognitive assessment in the Clinical Antipsychotic Trials of Intervention Effectiveness (CATIE) project schizophrenia trial: development, methodology, and rationale. AB - Patients with schizophrenia are severely impaired in crucial aspects of neurocognitive function. This impairment is the strongest clinical correlate of poor long-term outcome and adaptive dysfunction. Reports of neurocognitive enhancement with second generation antipsychotic medications have thus offered promise for improvement in the long-term outcome of patients with schizophrenia. However, the majority of these studies have had serious weaknesses in methodology, such as open-label design, small samples, or inappropriate dosing of medications. More recent studies have addressed these methodological issues but have been of short duration and have largely been sponsored by pharmaceutical companies. The Clinical Antipsychotic Trials of Intervention Effectiveness (CATIE) project is a unique opportunity to address the comparative neurocognitive effectiveness of available antipsychotic medications. This article describes the neurocognitive methods used in the schizophrenia trial of the CATIE project, including the selection and training of neurocognitive raters, patient inclusion criteria for assessment, rationale for the choice of neurocognitive instruments, and methodology for each neurocognitive test. PMID- 12908662 TI - Statistical approaches to effectiveness measurement and outcome-driven re randomizations in the Clinical Antipsychotic Trials of Intervention Effectiveness (CATIE) studies. AB - The design of the Clinical Antipsychotic Trials of Intervention Effectiveness (CATIE) schizophrenia and Alzheimer's disease studies pose several statistical challenges, including issues related to performing multiple comparisons, defining effectiveness outcomes, and collecting and analyzing data from a design with multiple outcome-driven re-randomizations. We discuss the CATIE strategy for addressing many hypotheses within the context of one clinical trial while controlling the overall type I error rate. We provide motivation for the use of two effectiveness outcomes: time to all-cause discontinuation and composite endpoints that combine outcomes from multiple domains, such as efficacy, safety, cost-effectiveness, and quality of life. Methods for statistical analysis of an outcome-driven re-randomization trial are compared and evaluated. We describe analysis within each phase, analysis based on the first randomization or treatment algorithms, and repeated measures modeling. Finally, strategies are described for designing an electronic data collection system for trials with repeated outcome-driven re-randomizations. PMID- 12908664 TI - The diffusion of new antipsychotic medications and formulary policy. AB - Spending on antipsychotic medications continues to escalate as new products such as atypicals are increasingly used to treat schizophrenia and other conditions. Given that per person spending on behavioral health benefits is shrinking while spending on pharmaceutical products is increasing, the psychotropic portion of mental health expenditures is likely to continue to increase in the future. The diffusion of these new behavioral health technologies, or the rate at which these products have spread through the market, has been very uneven. Differences in adoption and diffusion rates of psychotropic medications across insurance settings, geographic regions, or subpopulations defined by age, gender, or racial or ethnic groups have important implications for the quality of care received by persons with mental illnesses. This article reviews the evidence on the diffusion of antipsychotic medication and discusses the implications of formulary policies on diffusion, addressing the health care service and administrative context in which the Clinical Antipsychotic Trials of Intervention Effectiveness (CATIE) project is being conducted and which it is intended to inform. PMID- 12908663 TI - Changing environments and alternative perspectives in evaluating the cost effectiveness of new antipsychotic drugs. AB - This article examines the ways in which changes in the treatment environment and in measurement perspectives can affect the evaluation of cost-effectiveness of new medications. In three studies we reexamined data from a clinical trial of haloperidol and clozapine conducted from 1993 to 1996. The results of the studies are as follows: Study 1 found that clozapine treatment was associated with significantly reduced inpatient costs, and increased outpatient costs, suggesting that as systems use less inpatient care and more outpatient care, more effective medications may increase, rather than decrease, costs in sicker patients. Study 2 found that while provider assessments and standard measures favored clozapine over haloperidol, patient responses showed little evidence of a clinical advantage for clozapine and a less favorable side-effect profile. Study 3 found that while annual drug costs in the published trial were estimated to be dollars 4,545 for a full year of clozapine treatment, atypical antipsychotic costs in 2000 were estimated to range from dollars 1,254 to dollars 3,016 in the Department of Veterans Affairs system, and from dollars 2,221 to dollars 8,147 in the private sector. In conclusion, cost-effectiveness, as evaluated in studies like CATIE, will increasingly need to be tied to service system contingencies, environments, and evaluation perspectives. PMID- 12908665 TI - Generating evidence to inform policy and practice: the example of the second generation "atypical" antipsychotics. AB - The introduction of the second generation "atypical" antipsychotics has been heralded as a major advance in the treatment of schizophrenia and other psychotic disorders. Systematic reviews have revealed only modest advantages over conventional antipsychotics and uncertainty about long-term efficacy and safety, yet the second generation antipsychotic drugs have been widely accepted into clinical practice. Although the existing evidence of the benefits and harms of atypical antipsychotics can facilitate decision making about individual patients, the randomized evidence remains inadequate to make valid and fully evidence-based policy statements such as clinical practice guidelines that are designed to apply to groups of patients. Further large randomized trials are needed, but these require patients and clinicians to be in equipoise, or substantially uncertain, about alternative therapies. Premature clinical practice guidelines or expert opinion can lead to changes in clinical practice that make it difficult or impossible to conduct the required trials and are therefore a disservice to patients. PMID- 12908666 TI - Randomized controlled trials in evidence-based mental health care: getting the right answer to the right question. AB - The purpose of clinical research is to answer this question: Would a new treatment, when added to the existing range of treatment options available in practice, help patients? Randomized controlled trials (RCTs)--in particular, double-blind RCTs--have important methodological advantages over observational studies for addressing this question. These advantages, however, come at a price. RCTs compare treatments using a particular allocation rule for assigning patients to treatments (random assignment) that does not mimic real-world practice. "Favorable" results from an RCT indicating that a new treatment is superior to existing treatments are neither necessary nor sufficient for establishing a "yes" answer to the question posed above. Modeled on an experimental design, RCTs are expensive in time and money and must compare simple differences in treatments. Findings have a high internal validity but may not address the needs of the field, particularly where treatment is complex and rapidly evolving. Design of clinical research needs to take account of the way treatments are allocated in actual practice and include flexible designs to answer important questions most effectively. PMID- 12908667 TI - Implementation of evidence-based practices in state mental health systems: implications for research and effectiveness studies. AB - There is a huge gap between knowledge and practice, and between what is known through research and what is actually implemented in public mental health systems. In the past 2 decades, rigorous research has demonstrated the effectiveness of various treatments and interventions but these findings have not resulted in broad-based implementation. A major contention of this article is that while there is a growing body of knowledge related to evidence-based practices, there is a lack of understanding and research related to factors critical for implementation. This article reviews the current status of implementation of evidence-based practices in the public mental health system and identifies challenges and barriers related to their dissemination. Based on this analysis, this article proposes a research agenda that promotes the development of a science of implementation of evidence-based practices. PMID- 12908668 TI - Science to services: consumers need "real-world" science. AB - Over the past decade there has been a revolution in pharmacotherapy for schizophrenia and related disorders. The second generation, or atypical, antipsychotic medications have demonstrated efficacy and generally better side effect profiles. However, from the perspective of policy makers the higher costs associated with these newer medications leads to tough decisions regarding their continued use in light of an escalating fiscal crisis. For consumers, both persons with the illness and their family caregivers, the budgetary cutbacks leave many scrambling for answers to questions that most treatment (efficacy) studies were never designed to answer. "Should we oppose formulary restrictions on principle alone, or is there scientific data that can be relied on to inform our position?" On a more personal note, many are asking whether or not to switch to one of the newer medications and which medication would be best for them. Unlike CATIE, efficacy studies were never designed to answer such questions. In this article, we start by highlighting how CATIE will fill important gaps in translating the results of efficacy studies to effectiveness in the real world. Both the development of the CATIE methodology and the study design itself reflect what we will refer to as "real-world science": i.e., science that sheds light on effectiveness in vivo and can inform decisions consumers, clinicians, and policy makers are faced with day-to-day. We discuss CATIE in the context of the fiscal crisis hitting MEDICAID programs leading many policy makers to take the more expensive, atypical antipsychotics off the list of medications made available to patients. We argue that studies like CATIE will be highly informative and ultimately vital to policy makers wishing to create mental health policies that will succeed. Throughout, we highlight how CATIE, and real-world science more generally, are vital to consumers striving to find the medication(s) that works best for them. Given the organic research design process, which arguably relies on a fuller range of stakeholders than any study of its kind before, we remain hopeful that CATIE can succeed in generating an unprecedented amount of real world science that consumers can use. PMID- 12908669 TI - First person account: medicines are not enough. PMID- 12908670 TI - Psychological trauma and schizotypal symptoms. AB - In a sample of 75 women recruited from the community, we measured trauma/maltreatment history and symptoms of schizotypal personality disorder, using both questionnaire and interview measures. As hypothesized, individuals with histories of trauma/maltreatment had elevated levels of schizotypal symptoms. Among types of trauma/maltreatment, reported childhood neglect was especially strongly associated with schizotypal symptoms. Although posttraumatic stress disorder symptom severity, depression, dissociation, and difficulty identifying one's emotions were all associated with schizotypal symptoms, they could not account completely for the association between trauma/maltreatment and schizotypal symptoms. PMID- 12908671 TI - The multidimensional scale of independent functioning: a new instrument for measuring functional disability in psychiatric populations. AB - The Multidimensional Scale of Independent Functioning (MSIF) is a new instrument for rating functional disability in psychiatric outpatients. The MSIF differs from other disability rating scales by providing discrete ratings of (1) role responsibility, (2) presence and level of support, and (3) performance quality. The MSIF, which consists of a semistructured interview and detailed rating anchors, was validated in 114 psychiatric outpatients. The instrument had good criterion, discriminative, interrater, and construct validity. Correlations between comparable ratings on the Social Adjustment Scale II (SAS II) ranged from 0.78 to 0.86. Nevertheless, redundancy analysis using canonical correlation demonstrated that, although the two instruments overlap, the MSIF contains information that is not contained in the SAS II. Furthermore, there was only modest shared variance with conceptually non-overlapping subscales in the SAS II. Interrater reliability (intraclass correlation coefficients) ranged from 0.74 to 1.00 for global and subscale scores. MSIF subscales performed as expected with respect to external validators such as hours of employment, earned income, supported versus nonsupported employment and housing, and mainstream versus nonmainstream educational status. MSIF global ratings were modestly correlated with IQ and psychopathology ratings, consistent with reports in the literature. Construct validity, estimated using Cronbach's alpha coefficient, was 0.72. The MSIF is a promising new instrument designed to circumvent several limitations with existing functional outcome instruments for longitudinal studies, intervention research, and services research. PMID- 12908672 TI - Monozygotic twins exhibit numerous epigenetic differences: clues to twin discordance? AB - The goal of this pilot study was to explore the putative molecular mechanisms underlying the phenotypic discordance of monozygotic (MZ) twins. Thus, patterns of epigenetic DNA modification were investigated in the 5'-regulatory region of the dopamine D2 receptor gene (DRD2) in two pairs of monozygotic twins, one concordant and one discordant for schizophrenia. The bisulfite DNA modification based approach was used to fine-map methylated cytosines in DRD2 in genomic DNA extracted from lymphocytes. Numerous DNA methylation differences were identified in the analyzed region both within and between the pairs of MZ twins. "Epigenetic distances" between MZ twins were calculated and used for the comparison of twin DRD2 methylation profiles. It was detected that the affected twin from the pair discordant for schizophrenia was epigenetically "closer" to the affected concordant twins than to his unaffected MZ co-twin. Although the epigenetic analysis was conducted for only several hundred base pairs of DRD2, the fact that numerous studies identified nonuniform methylation patterns across the clones of bisulfite-modified DNA from the same individual, as well as nonuniform patterns across different individuals, argues for the universality of intra- and interindividual epigenetic variation. Epigenetic studies should provide insight into the molecular causes of differential susceptibility to a disease in genetically identical organisms that may generalize to singletons. PMID- 12908673 TI - Experimental manipulation of prior experience: Effects on item and associative recognition. AB - Frequency of exposure to very low- and high-frequency words was manipulated in a three-phase (familiarisation, study, and test) design. During familiarisation, words were presented with their definition (once, four times, or not presented). One week (Experiment 1) or one day (Experiment 2) later, participants studied a list of homogeneous pairs (i.e., pair members were matched on background and familiarisation frequency). Item and associative recognition of high- and very low-frequency words presented in intact, rearranged, old-new, or new-new pairs were tested in Experiment 1. Associative recognition of very low-frequency words was tested in Experiment 2. Results showed that prior familiarisation improved associative recognition of very low-frequency pairs, but had no effect on high frequency pairs. The role of meaning in the formation of item-to-item and item-to context associations and the implications for current models of memory are discussed. PMID- 12908674 TI - Ordering our world: An examination of time in autobiographical memory. AB - In two studies people judged the order in which two real-world events occurred. Ordering performance was better for events that were recent or widely separated in time. Ordering performance was also consistently related to predicted event memorability and to the amount of processing given to an event during encoding. Ordering performance was not consistently related to the person-typicality, pleasantness, or emotional intensity of the events, and was also not related to whether the two events judged came from the same thematic category. These results suggest that memory for event order is not entirely reconstructed from event content. We suggest that the self-concept may sometimes serve as an implicit timekeeper in autobiographical memory. PMID- 12908675 TI - The Prospective and Retrospective Memory Questionnaire (PRMQ): Normative data and latent structure in a large non-clinical sample. AB - The Prospective and Retrospective Memory Questionnaire (PRMQ; Smith, Della Sala, Logie, & Maylor, 2000) was developed to provide a self-report measure of prospective and retrospective memory slips in everyday life. It consists of sixteen items, eight asking about prospective memory failures, and eight concerning retrospective failures. The PRMQ was administered to a sample of the general adult population (N = 551) ranging in age between 17 and 94. Ten competing models of the latent structure of the PRMQ were derived from theoretical and empirical sources and were tested using confirmatory factor analysis. The model with the best fit had a tripartite structure and consisted of a general memory factor (all items loaded on this factor) plus orthogonal specific factors of prospective and retrospective memory. The reliabilities (internal consistency) of the Total scale and the Prospective and Retrospective scales were acceptable: Cronbach's alpha was 0.89, 0.84, and 0.80, respectively. Age and gender did not influence PRMQ scores, thereby simplifying the presentation and interpretation of normative data. To ease interpretation of scores on the PRMQ, tables are presented for conversion of raw scores on the Total scale and Prospective and Retrospective scales to T scores (confidence limits on scores are also provided). In addition, tables are provided to allow users to assess the reliability and abnormality of differences between an individual's scores on the Prospective and Retrospective scales. PMID- 12908676 TI - A cross-modal weapon focus effect: The influence of a weapon's presence on memory for auditory information. AB - To examine whether a weapon's presence impairs witnesses' memory for auditory information (as it impairs memory for visual information), we conducted two experiments in which undergraduates watched one version of a videotape depicting a male target who held either a weapon or a neutral object and conversed with a female character. The semantic content of his remarks was either easy or difficult to comprehend. The weapon's presence did not affect voice identification accuracy or memory for the target's vocal characteristics (e.g., pitch, loudness, speech rate) but did worsen memory for semantic content in the Difficult Comprehension condition. Our results can be explained by multiple resource models of attention, which propose separate resource "pools" for different sensory modalities. PMID- 12908677 TI - Long-term flashbulb memory for learning of Princess Diana's death. AB - Flashbulb memories are unusually vivid recollections of the circumstances in which one first learns of a shocking and emotionally arousing event. In the present study we examined students' memories concerning the death of Diana. Princess of Wales. Participants completed a standard flashbulb memory questionnaire 1 week after her death, in which they reported the circumstances in which they first heard the news. Accuracy of these memories (defined by consistency of responses across delays) was investigated by either a single retest 18 months after the event, or two retests at 3 and 18 months after the event. Participants' descriptions remained accurate at both 3 and 18 months, and testing at 3 months had no effect on accuracy of the subsequent 18-month responses. Consistent with previous research, both emotional intensity and rehearsal were related to accuracy. PMID- 12908678 TI - Posthypnotic amnesia for a first romantic relationship: Forgetting the entire relationship versus forgetting selected events. AB - This experiment investigated the impact of suggestion focus on posthypnotic amnesia (PHA) for memories of a first romantic relationship. During hypnosis, high and low hypnotisable participants recalled specific memories from this period in response to 10 cue phrases (Elicitation). They then received a PHA suggestion that targeted either the entire period or specific memories from that period. Participants' explicit memory was indexed by cued recall after PHA was administered (memories recalled to "old" and "new" cues; Recall 1) and after it was cancelled (Recall 2). A social judgement task indexed dissociations between implicit and explicit memory. PHA had the greatest impact on highs', but not lows', memory performance (in terms of memories recalled to old cues, recall latency, and qualitative memory ratings) when the suggestion targeted the entire period rather than specific events. We discuss these findings in terms of the parameters of PHA's influence on memory, its value for exploring the nature and structure of autobiographical memory, and its utility as a laboratory analogue of functional amnesia. PMID- 12908679 TI - Memory for a staged criminal event witnessed live and on video. AB - Memory for a staged robbery was tested in two groups of participants witnessing the event either live (n = 62) or on video (n = 64). Immediately after the event participants filled out a questionnaire probing memory with emphasis on the timing of the event and robber characteristics. The results showed that participants who watched a video recording of the event reported more details and with a higher accuracy than participants who were present on the scene, but the pattern of memory errors were similar in the two conditions. It is concluded that laboratory experiments may overestimate the memory of eyewitnesses but are otherwise able to simulate essential aspects of memory performance in naturalistic contexts. PMID- 12908680 TI - Endothelium, nitric oxide, and anesthetics. PMID- 12908681 TI - Vital capacity induction with 8% sevoflurane and N2o causes cerebral hyperemia. AB - PURPOSE: Little is known about the influence of high-dose sevoflurane on cerebral volume. We evaluated induction time and cerebral blood volume with 8% sevoflurane using the "vital capacity induction" technique. METHODS: Thirty-four patients were randomly allocated into three groups. Group P received 2.0 mg x kg(-1) of propofol i.v. and inhalation of 67% N2o/O2, whereas group S5 and group S8 received inhalation of primed 5% and 8% sevoflurane in 67% N2O/O2, respectively. Induction time was measured as the time from the start of inhalation, or from the end of injection, until loss of eyelash reflex. Near-infrared spectroscopy and bispectral index (BIS) were monitored continuously until 3 min after tracheal intubation. RESULTS: Induction time was less in group S8 (17.3 +/- 6.4s, mean +/- SD) than in groups P (25.7 +/- 8.2s) and S5 (33.0 +/- 16.8s). There was a significant increase in cerebral blood volume after intubation in group S8, as suggested by higher cerebral oxyhemoglobin and total hemoglobin levels. There were no differences in BIS scores among the groups during the study period. CONCLUSION: Vital capacity inhalation of 8% sevoflurane produces a faster loss of eyelash reflex than does 5% sevoflurane or propofol, but increases cerebral blood volume. PMID- 12908682 TI - Survey of anesthesia practice in spine surgery patients in the United States. AB - PURPOSE: There is a wide range of anesthetic practice in spine surgery with respect to anesthetic choice, blood conservation, and monitoring. There is no ideal technique with respect to each of these choices. This study was designed to determine the usual practice for members of the Society of Neurosurgical Anesthesia and Critical Care (SNACC), with regard to the use of anesthetic technique, the type of monitoring used (and the person responsible for its application and interpretation), and the blood conservation strategies most often utilized for the management of patients undergoing spinal surgery. METHODS: A total of 415 questionnaires were mailed to all current members of the SNACC. Of these, 85 (23%) were completed and returned. RESULTS: The majority of the respondents administer general anesthesia for lumbar laminectomy. Somatosensory evoked potential (SSEP) monitoring is available at most of the institutions (94%) but only utilized in 75% of patients undergoing a Harrington rod placement. Motor evoked potential monitoring is available at 50% of the institutions, but utilized 25% of the time for these surgeries. The two most frequent blood conservation strategies utilized are intraoperative salvage and autologous donation. CONCLUSION: The most frequent monitoring utilized for major spinal surgeries is SSEP. Autologous donation and intraoperative salvage are the most frequent blood conservation methods utilized. PMID- 12908683 TI - Effects of halothane and isoflurane on acetylcholine-induced, endothelium dependent vasodilation in perfused rat mesenteric arterial beds. AB - PURPOSE: The present study was designed to examine the effects of halothane and isoflurane on acetylcholine-induced, endothelium-dependent vasodilation in rat mesenteric arterial beds perfused at a constant flow both in vitro and in situ. METHODS: In the in-vitro preparation, the mesenteric artery was cannulated and perfused (5 ml x min(-1)). The perfusion pressure was continuously monitored. Under active tone induced by methoxamine, the effects of halothane and isoflurane on the vasodilator response to acetylcholine in either the presence or absence of NG-nitro-L-arginine (L-NA), tetraethylammonium (TEA), or KCl (30 mM) depolarization were examined. All experiments in these preparations were performed in the presence of indomethacin (10 mM). In the in-situ experimental model, rats were anesthetized with pentobarbital and the lungs were mechanically ventilated via a tracheostomy with a ventilator. The superior mesenteric artery was cannulated and used for the monitoring of the perfusion pressure. Blood shunting with constant flow (2 ml x min(-1)) from the carotid artery to the superior mesenteric artery was introduced with clamping at the immediately distal portion of the mesenteric artery branching. Following 20-min ventilation with halothane or isoflurane at 1 minimum alveolar concentration (MAC) in oxygen, acetylcholine was given from the mesenteric artery, under active tone induced by norepinephrine (100 mg x kg(-1) x hr(-1)). RESULTS: In the in-vitro preparation, the nitric oxide synthase inhibitor, L-NA (100 microM) did not affect vasodilations to acetylcholine (1, 10 nM), while the K+ channel inhibitor TEA (10 mM), as well as KCl (30 mM), significantly reduced these vasodilations. However, only in the presence of L-NA, TEA and KCl completely abolished the vasodilations produced by acetylcholine. The higher concentrations of halothane (2.0%, 3.0%), but neither isoflurane (3.0%) nor the lower concentration of halothane (1.0%), significantly impaired vasodilator responses to acetylcholine in the presence of L-NA, whereas the volatile anesthetics did not affect these vasodilations in the absence of L-NA. Halothane (2.0%) did not alter the vasodilation produced by acetylcholine in the presence of TEA or KCl. In the in-vivo preparation, the vasodilator effects of acetylcholine (1 and 10 nmol) were not affected by the inhalation of halothane (1.0%) or isoflurane (1.3%). CONCLUSION: These results suggest that, in resistance arteries in conditions of constant flow, halothane and isoflurane do not affect vasodilations in response to an endothelium dependent agonist. However, in these preparations, once the enzymatic activity of nitric oxide synthase is inhibited, higher concentrations of halothane, but neither isoflurane nor the lower concentration of halothane, appear to impair endothelium-dependent relaxations, probably mediated by TEA-sensitive K+ channels. PMID- 12908684 TI - Hypotension associated with systemic aggregated anaphylaxis is not attenuated by a selective endothelin-A receptor antagonist, BQ 610, in rabbits in vivo. AB - PURPOSE: The present study was done to investigate the role of endothelin-1 (ET 1) in hypotension and bronchospasm provoked by anaphylaxis in rabbits in vivo. METHODS: Forty-five rabbits sensitized to horse serum were randomly allocated to five groups: Group 1 (n = 10) received 0.5 nmol x kg(-1) of ET-1; Group 2 (n = 10) received 0.5 nmol x kg(-1) of ET-1 and 200 nmol x kg(-1) of a selective ETA receptor antagonist, BQ 610, without anaphylaxis; Group 3 (n = 5) received 200nmol x kg(-1) of BQ 610 alone without anaphylaxis, Group 4 (n = 10) received normal saline alone before being antigen challenged to induce anaphylaxis; Group 5 (n = 10) received 200 nmol x kg(-1) of BQ 610 before antigen challenge. RESULTS: Mean arterial pressure (MAP) values were significantly different between Groups 1 and 2. Heart rate (HR), central venous pressure (CVP), dynamic pulmonary compliance (C(dyn)), and pulmonary airway resistance (R(L)) did not differ significantly between Groups 1 and 2. MAP values were significantly decreased compared with baseline in both Groups 4 and 5; however, the values were not significantly different between two groups. CVP values were significantly different between Groups 4 and 5 only at the 15-min time point following antigen challenge. HR, R(L), and C(dyn) values were not significantly different between Groups 4 and 5, nor were the survival rates. CONCLUSION: BQ 610 does not improve hypotension or survival rates in systemic aggregated anaphylactic rabbits in vivo, implying that circulating ET-1 may not play an important role in anaphylaxis, although direct proof of production of circulating ET-1 or activation of ETA receptors is lacking in this study. PMID- 12908685 TI - Recent advances in obstetric anesthesia. AB - The low-dose technique of combined spinal/epidural analgesia is to be welcomed in obstetrics. Its merits include rapid onset of analgesia, with the flexibility of an epidural technique, and high maternal satisfaction. It is a safe and effective technique. Pulse oximetry should be employed when using intrathecal opioids. Commercially available combined-needle devices may make this technique more attractive to users. The role of spinal anesthesia for emergency cesarean section in severe preeclampsia has been reevaluated recently. We consider it a feasible option for those severely preeclamptic women requiring urgent cesarean section who do not have an epidural catheter in place. The choice of anesthetic technique for this patient population should be made on clinical judgment and not on anticipated hemodynamic changes. Spinal anaesthesia for cesarean section is associated with hypotension; however, certain interventions may reduce the incidence and severity of the hypotension. An increase in cardiac output appears to be key in attenuating the hypotensive response to spinal anesthesia. Colloids have exhibited most success in this regard. At our institution, we do not delay spinal anesthesia for urgent cesarean section in order to administer a predetermined volume of fluid; in such cases, we simultaneously administer a fluid preload and spinal anesthesia. Recent studies regarding the use of cell savers for blood conservation in obstetrics are based on small numbers of patients. These studies show great promise, particularly with the modern emphasis on avoiding blood transfusion, which can be massive in this usually young patient population. PMID- 12908686 TI - One hundred years of Nobel Prizes and 150 years of anesthesiology. PMID- 12908687 TI - Acute airway obstruction secondary to retropharyngeal hematoma. PMID- 12908688 TI - Perioperative management of lobectomy in a patient with hypertrophic obstructive cardiomyopathy treated with dual-chamber pacing. PMID- 12908689 TI - Seizure induced by a small dose of fentanyl. PMID- 12908690 TI - Postherpetic neuralgia as a risk factor for classic heatstroke. PMID- 12908691 TI - Modified insertion technique of the laryngeal mask airway in children: a comparison with standard technique. PMID- 12908692 TI - Epidural spinal cord stimulation for neurogenic bladder. PMID- 12908693 TI - Baroreflex sensitivity and hemodynamic changes in elderly and young patients during propofol anesthesia. PMID- 12908695 TI - Usefulness of ultrasonic two-way Doppler flow detector in percutaneous arterial puncture in patients with hemorrhagic shock. PMID- 12908694 TI - Effect of infraorbital nerve block under general anesthesia on consumption of isoflurane and postoperative pain in endoscopic endonasal maxillary sinus surgery by Higashizawa and Koga. PMID- 12908696 TI - A minimally invasive technique for aggressively monitoring high-risk patients undergoing low-risk surgical procedures that do not require endotracheal intubation. PMID- 12908697 TI - Small particulate gadolinium oxide and gadolinium oxide albumin microspheres as multimodal contrast and therapeutic agents. AB - RATIONALE AND OBJECTIVE: To prepare and characterize new particulate contrast media, small particulate gadolinium oxide (SPGO) and gadolinium oxide albumin microspheres (GOAM), as prototype multimodal imaging and therapeutic agents. METHODS: SPGO was purchased from Alfa Aesar Inc. (Ward Hill, MA). GOAM were synthesized via ultrasonic irradiation using SPGO and 5% bovine serum albumin in aqueous solution. SPGO and GOAM were characterized by size, concentration, structure, and gadolinium content. Their relaxivity at high magnetic field strength and x-ray attenuating abilities were evaluated in 0.4% agar gel at room temperature. RESULTS: SPGO were confirmed to be 20-40 nm in diameter. GOAM have an average size of 2 to 5 microm and show a relatively homogeneous distribution of SPGO within the albumin microspheres. T1 and T2 relaxivity of GOAM was 6.7 seconds(-1) mmol/L(-1) and 38.5 seconds(-1) mmol/L(-1), respectively, while that of SPGO was 0.2 seconds(-1) mmol/L(-1) and 6.8 seconds(-1) mmol/L(-1), respectively. At 0, 0.004, 0.16, 4.0, and 16.0 mmol/L SPGO, x-ray attenuation values were measured at 2.11, 3.48, 7.06, 39.83, and 187.33 Hounsfield units, respectively. CONCLUSIONS: Use of SPGO and SPGO encapsulated in non-heat-hardened albumin microspheres (GOAM) represents a new approach. SPGO and GOAM have suitable physicochemical properties to warrant further biophysical and animal studies and reevaluation of toxicity limitations. PMID- 12908698 TI - Comparison of different types of blood pool agents (P792, MS325, USPIO) in a rabbit MR angiography-like protocol. AB - RATIONALE AND OBJECTIVES: The objective of this study is to determine the influence of the pharmacokinetic behaviors of different classes of blood pool agents (BPA) on a rabbit experimental model that mimics a magnetic resonance angiographic protocol. BPA were as follows: P792, a macromolecular agent (RCBPA), USPIO, an ultrasmall superparamagnetic iron oxide particle agent (SCBPA), and MS 325, a small gadolinium chelate that expresses intravascular behavior by reversible albumin binding. METHODS: The 2 main phases of early distribution following contrast agent injection, that is, the bolus phase and the steady-state phase, are investigated by measuring Gd or Fe blood concentrations in the first 5 minutes postinjection. T1 relaxation times and r1 relaxivity were calculated at each time point of blood sampling. Furthermore, in the case of MS-325, the concentrations of the free and bound forms were calculated, according to the measured concentrations and the apparent r1 relaxivities. RESULTS: Injected under similar conditions, the 3 BPA have, during the bolus phase, a comparable profile to Gd-DOTA. Signal enhancement was maximum during this short bolus phase, as were the T1 relaxation times under 30 ms for all agents. At 1 minute postinjection, P792 (r1 = 39 seconds(-1) x mmol/L(-1), 20 MHz) demonstrated the same pharmacokinetic behavior as USPIO (r1 = 33 seconds(-1) x mmol/L(-1), 20 MHz): C1 minute/C0 values were 91 +/- 6% and 92 +/- 12%, respectively. Immediately after the injection at clinical dose, 74% of MS-325 was in free form, resulting in an apparent r1 relaxivity of only 13 seconds(-1) x mmol/L(-1) (20 MHz); 1 minute postinjection, the C1 minute/C0 value of 61 +/- 4% was the lowest as compared with P792 and USPIO and the bound form represented 75% of the MS-325 molecules. CONCLUSIONS: The BPA P792 and USPIO have favorable properties that result from their intravascular retention and their lack of extravasation, allowing optimal contrast between the vessel and the adjacent tissue for several minutes postinjection. Combining a rapid body clearance and a marked T1 effect, P792 presents optimal blood pool characteristics for angiographic applications. During the bolus phase, MS-325 is mainly in free form, which presents the disadvantage of increasing the tissue signal background, due to extravasation of the free form. PMID- 12908699 TI - Development of new glucosylated derivatives of gadolinium diethylenetriaminepentaacetic for magnetic resonance angiography. AB - RATIONALE AND OBJECTIVES: A possible approach for the extension of the vascular residence time of contrast agents relies on the renal reabsorption mechanisms of some molecules such as glucose. In this study, various small-molecular-weight glucosyl derivatives of gadolinium diethylenetriaminepentaacetic (Gd-DTPA) were synthesized and their vascular half-life was studied. METHODS: Several Gd-DTPA bisamides carrying glucosyl groups bound by different linkers were prepared. The pharmacokinetics and biodistribution of these compounds were determined on Wistar rats. RESULTS: The sugar moieties linked to Gd-DTPA efficiently reduce the renal excretion of some derivatives. The interaction with renal carrier has not been clearly demonstrated, nor was any interaction observed with blood components. CONCLUSIONS: Two of the new glucosylated derivatives of Gd-DTPA (Cd-DTPA-BC2-beta cellobionA 2 and Gd-DTPA-BC4-beta-glucosylA 7) can be proposed as blood-pool MR contrast agents, considering their vascular remanence. PMID- 12908700 TI - Helium3 polarization using spin exchange technique: application to simultaneous pulmonary ventilation/perfusion imaging in small animals. AB - RATIONALE AND OBJECTIVES: To develop a simple and robust helium3 polarization system dedicated to small animal imaging. To demonstrate the potential of helium3 imaging for pulmonary ventilation and perfusion studies. METHODS: A home-built polarization system based on spin-exchange technique was used. This system was applied for magnetic resonance imaging ventilation studies on rats using a 2-T magnet. Projection-reconstruction sequences combined with the sliding-window technique were used for acquisition of high temporal resolution ventilation images. RESULTS: Helium3 polarization levels up to 25% were obtained. Simultaneous ventilation and lung perfusion images were acquired with intravenous injection of superparamagnetic contrast agents. Dose effects were investigated using several contrast agent concentration values. CONCLUSIONS: A tabletop helium3 polarization system was realized. This equipment, which is easy to operate, allows the production of polarized gas appropriate to the requirements of small animal studies. This polarization system was used successfully on a ventilation/perfusion imaging study using intravascular contrast agent. PMID- 12908701 TI - Distribution of ventilation in lung transplant recipients: evaluation by dynamic 3He-MRI with lung motion correction. AB - RATIONALE AND OBJECTIVES: The ability of motion corrected dynamic 3He-magnetic resonance imaging (MRI) to discriminate distributional patterns of inhaled hyperpolarized 3He between different groups of lung transplant recipients was evaluated. METHODS: An ultrafast low-angle shot 2D sequence (temporal resolution 128 ms) was used for ventilation 3He-MRI of 11 single and 6 double lung transplant recipients. After digital motion correction, signal kinetics were evaluated in a tracheal and 7 pulmonary regions of interest. Results from grafts and native lungs as well as from normal and rejected grafts were compared with each other and to reference values from healthy subjects. RESULTS: In emphysema patients, median alveolar rise time, a parameter for increase of alveolar signal, was 0.28 seconds for the graft and 0.48 seconds for the native lung, in fibrosis patients its median was 0.46 seconds for the graft and 0.21 seconds for the native lung. In double lung recipients, alveolar rise time was 0.29 seconds in normal and clinically rejected grafts. CONCLUSIONS: Dynamic ventilation 3He-MRI discriminated normal lung grafts from diseased native lungs in single lung recipients. Graft rejection in double lung recipients could not be discriminated. PMID- 12908702 TI - Visualization of normal and interrupted lymphatic drainage in dog legs with interstitial MR lymphography using an extracellular MR contrast agent, gadopentetate dimeglumine. AB - PURPOSE: The capability of the interstitial magnetic resonance lymphography (MR LG) using a widely available extracellular contrast agent, gadopentetate dimeglumine, for the visualization of normal and interrupted lymphatic drainage was tested in dog legs. MATERIALS AND METHODS: With a 7-day interval, 1 mL and 2 mL undiluted gadopentetate dimeglumine were injected intradermally into the dorsal foot of both hind legs of 12 dogs. T1-weighted 3-dimensional (3D) spoiled gradient-echo (repetition time/echo time = 6.7/1.6 ms) and maximum intensity projection (MIP) images covering the legs and pelvic region were acquired at 1.5 T for 10 minutes after 30 seconds gentle massage at the injection sites. These dogs also underwent the MR-LG with 2 mL of this contrast agent 7 days after surgical ligation of the popliteal lymphatic vessels. RESULTS: In the normal dogs, the interstitial MR-LG quickly and consistently visualized the direct connection of the lymphatic vessels and lymph nodes draining from the injection sites up to the inguinal region, without any late adverse effects. The enhancement effect was significantly greater with a 2-mL dose than with a 1-mL dose. After lymphatic obstruction, the MR-LG clearly revealed the anatomic compromises of the lymphatics, such as poor enhancement of the affected lymphatic vessels and lymph nodes, collateral lymphatic vessels, abnormal soft-tissue enhancement, and delayed lymphatic migration of the contrast agent. The topographic 3D images provided a comprehensive anatomy of these normal and compromised lymphatic drainage. CONCLUSION: Although gadopentetate dimeglumine is not lymphotropic, the interstitial MR-LG using this T1-contrast agent appears to have potential for quick and sufficient mapping of the lymphatic drainage from the injection sites and for the characterization of interrupted lymph flow. PMID- 12908703 TI - Sentinel node detection using contrast-enhanced power Doppler ultrasound lymphography. AB - RATIONALE AND OBJECTIVES: To establish the feasibility of using contrast-enhanced interstitial ultrasound (US) lymphography as an alternative to current sentinel node detection methods. METHODS: Aqueous US contrast microbubble suspensions of varying diameter were evaluated in vitro to characterize response to insonation. Contrast media were then injected subcutaneously into the distal extremities of 11 normal dogs to target the cervical and popliteal lymph nodes (nodes, n = 40). First-order (sentinel) lymph nodes and second-order sublumbar nodes were imaged intermittently from 0 to at least 120 minutes following contrast injection using continuous power Doppler mode. Lymphoscintigraphy studies were performed on 4 dogs to verify lymphatic drainage patterns and sentinel lymph nodes. RESULTS: Contrast enhancement occurred in 34/40 (85%) sentinel nodes overall and in 30/32 (94%) nodes when submicron or near-micron diameter bubble formulations were used. In many instances, enhancement persisted throughout the imaging period. Contrast response was most pronounced using a high mechanical index and tissue artifact was reduced or eliminated when using a high pulse repetition frequency. CONCLUSIONS: Contrast-enhanced interstitial US lymphography could serve as an alternative to current sentinel node detection methods. Preliminary findings suggest that submicron or near-micron-diameter bubbles may be suitable for lymphatic imaging applications. PMID- 12908705 TI - Emerging issues in multidetector computed tomography imaging. PMID- 12908704 TI - Calcium-dependent injury of human microvascular endothelial cells induced by a variety of iodinated radiographic contrast media. AB - RATIONALE AND OBJECTIVES: The aim of the present study was to determine the possible mechanisms underlying the endothelial cell damage induced by iodinated radiographic contrast materials (RCM). METHODS: The cultured human skin microvascular endothelial cells (HMVECs) were exposed to various contrast media, and the cell viability was measured by mitochondrial enzyme activity. Nuclear damage was assessed by Hoechst 33342 staining and a fluorescent single-cell gel electrophoresis. The effects of contrast materials on the cellular ATP content and intracellular free Ca2+ concentration were subsequently examined. RESULTS: Although the iodinated RCM tested all caused the cell injury in HMVECs, ionic RCM including amidotrizoate and ioxaglate were more potent in producing the cell damage than nonionic RCM. It is unlikely that the contrast material-induced cell damage is associated with hyperosmolality, since hyperosmolar solution of mannitol or NaCl had no marked influence on the endothelial cell viability. Nuclear damage was noted in cells exposed to amidotrizoate. Amidotrizoate lowered cellular ATP content while elevating the intracellular free Ca2+ concentration. It was notable that the RCM-induced endothelial cell damage was reversed by the chelation of intracellular Ca2+ with 1,2-bis(o-aminophenoxy)ethane-N,N,N',N' tetra-acetic acid but not by the removal of extracellular Ca2+. CONCLUSIONS: Both ionic and nonionic contrast materials caused nuclear damage of endothelial cells. The decrease in tissue ATP content and elevation of intracellular Ca2+ are likely to contribute to the contrast materials-induced endothelial cell damage. PMID- 12908706 TI - Contrast optimization and scan timing for single and multidetector-row computed tomography. AB - Various complex pharmacokinetic interrelationships affect the use of contrast media for computed tomography (CT) imaging. These include factors related to each patient's unique body habitus and their degree of health, such as their age, gender, height, weight, and cardiovascular status. Of equal importance are factors related to the contrast material injection. The volume, concentration, and rate of injection all affect the degree of enhancement that is achieved with an injection of contrast material. In addition, the injection technique--whether the contrast is infused at a constant injection rate (uniphasic injection) or whether the rate is altered during the injection (multiphasic injection)--also affects the magnitude and duration of contrast enhancement. In body CT imaging, the liver poses unique challenges in managing the use of intravenous contrast material because of its dual blood supply and the need to complete imaging before equilibrium occurs between the intravascular and extravascular compartments. The magnitude of hepatic enhancement that is ultimately achieved is related primarily to the amount of iodinated contrast material that accumulates in the extravascular space within the target organ, independent of the speed of the CT scanner. The key determinant of the onset of the equilibrium phase is the injection duration. Given that a high injection flow rate (4-5 ml/s) is desirable for arterial phase imaging, the injection duration is maintained by using an appropriate contrast volume. Thus, modifications of the total iodine dose are best performed by altering the contrast concentration. Use of a high contrast concentration (400 mg iodine/ml) may be advantageous in heavy patients, or in patients in whom routine imaging is married with a need for high-detail imaging of the vasculature with high resolution CT angiography. The magnitude of arterial enhancement that is achieved is related to both the concentration of contrast and the rate of administration. The speed of the scanner determines its ability to record image data during the most advantageous time period, the peak of arterial enhancement. Thus, rapid imaging is particularly advantageous for optimal contrast use in CT angiography as well as in multiphasic imaging of the parenchymal organs. PMID- 12908707 TI - Liver imaging with multidetector helical computed tomography. AB - The speed and flexibility of multidetector computed tomography (MDCT) have led to improvements in liver imaging, particularly related to the detection and characterization of focal lesions. This report discusses the different phase of liver enhancement following the bolus administration of iodinated contrast material, and the enhancement pattern of various liver lesions during these phases. We also propose guidelines for designing protocols for MDCT of the liver and discuss the principles of contrast media delivery. PMID- 12908708 TI - Helical computed tomography of the pancreas: potential impact of higher concentrated contrast agents and multidetector technology. AB - The introduction of multidetector helical computed tomography (CT) has revolutionized the field of pancreatic imaging and has created a new dimension of temporal and spatial resolution. The speed of multidetector CT can either be used to reduce the time needed to cover a given volume or to increase the resolution along the z-axis, thereby resulting in nearly isotropic voxels. Pancreatic perfusion phases can thus be more clearly differentiated and small structures are better delineated, leading to an improvement in the diagnostic value of pancreatic CT. Contrast media with a higher iodine concentration seem to be superior to lower concentrated contrast agents, mainly because of the better evaluability of the vessels in the arterial examination phase. This manuscript reports current CT imaging strategies for patients with pancreatic cancer with emphasis on vascular invasion, lymph node metastasis, liver metastasis, and peritoneal carcinomatosis. PMID- 12908709 TI - Computed tomography angiography: principles and clinical applications. AB - Clinical applications of computed tomography (CT) angiography have increased with the improved technology of multidetector CT systems. Adequate contrast enhancement and the timing of image acquisition are key elements in producing technically adequate CT angiograms. This review article provides guidelines and protocols for four-, eight- and 16-channel multidetector systems in studies of the thoracoabdominal aorta, aortoiliac, and abdominal visceral vasculature, abdominal and extremity run-off studies, and carotid/cerebral CT angiography. PMID- 12908710 TI - Benjamin Gompertz revisited. PMID- 12908711 TI - Variant Creutzfeldt-Jakob disease. AB - A variant form of Creutzfeldt-Jakob disease (vCJD) has had major impact in Europe during the last decade. In this article, we review the aetiology of vCJD and its relation with bovine spongiform encephalopathy. Further, treatment of the disease, the strategies focusing on prevention of transmission, future prospects, and the limitations encountered in epidemiological research of vCJD are discussed. PMID- 12908712 TI - The validity of self-reported leisure time physical activity, and its relationship to serum cholesterol, blood pressure and body mass index. A population based study of 332,182 men and women aged 40-42 years. AB - The importance of leisure time physical activity as a health indicator became more obvious after the results of large prospective studies were published. The validity of these results depends upon both the selection of the active individuals and to what extent self-reported physical activity reflects the individual's true activity. The purpose of this paper is to describe the changes in self-reported physical activity, and to assess the relation between this variable and other biological risk factors such as blood lipids, blood pressure and body mass index (BMI). This report also aims at corroborating the validity of self-reported physical activity by assessing the consistency of the associations between these biological risk factors and physical activity during a 25-years period. The basis for this analysis is a long lasting observational study with a questionnaire as the most important research instrument, in addition to physiological and biological factors such as BMI, blood pressure and blood lipids. The study population consists of 332,182 individuals, aged 40-42 from different counties in Norway who were invited to participate in health survey during 1974-1999. The objectives of this study are (1) to describe changes in self-reported physical activity from 1974 to 1999; (2) to assess the relation between physical activity and the biological variables; and (3) to corroborate the validity of the variable physical activity by assessing the consistency of the above analysis. The results of the analyses of association between decade of birth and self-reported physical activity show that physical activity among 40 aged individuals decreased during 1974-1999. This trend is stronger among the men. Multivariate analyses revealed differences in BMI and serum cholesterol between levels of self-reported physical activity, gender, smoking habits and decade of birth. The explained percentage of the total variance ranged from 6% for BMI to 7% for serum cholesterol. The similar shape of serum cholesterol and BMI according to physical activity indicates that the validity of self-reported physical activity has remained stable over these 25 years. Furthermore, the analysis of covariance showed that the slopes relating year of birth and serum cholesterol and BMI are parallel for self-reported physical activity thus the validity of the variable is confirmed. PMID- 12908713 TI - Serum cotinine as a marker of environmental tobacco smoke exposure in epidemiological studies: the experience of the MATISS project. AB - To describe serum cotinine levels in a rural Italian population and to examine its usefulness as an epidemiologic biomarker of nicotine exposure, cross sectional data collected in 1993 for the MATISS Project (2098 men and 1352 women, aged 20-79 years) were used. The study population consisted of 977 current smokers, 882 nonsmokers reporting exposure to environmental tobacco smoke (ETS) and 1520 nonsmokers reporting no ETS exposure. Mean values of serum cotinine measured by radioimmunoassay for never smokers, ex-smokers and current smokers (including four categories of cigarette consumption), and for categories of ETS exposure in all nonsmokers were calculated. In univariate analysis, there was a positive association between self-reported nicotine exposure and serum cotinine levels in all groups. Using self-reported status as truth, sensitivity and specificity for various cotinine cutoff points were estimated to distinguish nonsmokers from smokers. The value of 15 ng/mL represented the best combined levels of sensitivity (95%) and specificity (96%). Using this cutoff point, the overall misclassification rate for self-reported nonsmokers was 2.1% and about two times greater for the more vs. the less educated. In multivariate analysis, reported ETS exposure among nonsmokers was significantly associated with serum cotinine even after adjusting for age, socio-demographic and behavioural factors, though the strength of the association was not strong. In conclusion, serum cotinine represents a reliable epidemiological marker of nicotine intake and may be helpful when studying ETS exposure. Improved information collection is needed to reduce misclassification among nonsmokers and enhance our understanding of the relationship between ETS and cotinine measures. PMID- 12908714 TI - Assessment of alcohol consumption by mailed questionnaire in epidemiological studies: evaluation of misclassification using a dietary history interview and biochemical markers. AB - BACKGROUND: Self-reported information on alcohol from questionnaires is generally assumed to introduce misclassification of consumption, mainly in the direction of underestimation. The aim of this study was to evaluate self-reported information on alcohol consumption from a mailed questionnaire by comparing to a dietary history interview and biochemical markers of alcohol intake. SUBJECTS AND METHODS: For 76 male twin pairs of the Finnish Twin Cohort Study aged 40-70 years information on self-reported alcohol consumption was collected through mailed questionnaire and dietary history interview. Carbohydrate-deficient transferrin (CDT), Gamma-glutamyltransferase (Gamma-GT) and mean corpuscular volume (MCV) were determined from blood samples. RESULTS: Mean levels of CDT, gamma-GT and MCV showed a rise with increased self-reported alcohol consumption already at low levels of reported consumption (<20 g alcohol/day). There was a positive correlation between reported amount alcohol intake per day and levels of CDT (r = 0.46), gamma-GT (r = 0.32) and MCV (r = 0.36) but within the high consumption group (> or = 30 g/day) there was no such correlation. The questionnaire had sensitivity of 28-43% and specificity of 89% for identification of high consumers of alcohol using the biochemical markers as reference and sensitivity 41% and specificity 94% using the dietary history interview as reference. Sensitivity was improved when information on binge drinking (82%) or possible drinking problems (73%) was considered. CONCLUSION: Comparison to dietary history interview as well as to biochemical markers indicate that self-reported information on alcohol consumption from a mailed questionnaire may be used to distinguish between groups with different levels of alcohol consumption. The suggested misclassification of high consumers implies that only strong associations between high alcohol intake and disease are likely to be detected in studies based on questionnaire data. PMID- 12908715 TI - On the relationship between human all-cause mortality and age. AB - Many equations can be used to study the relationship between mortality rates and age: Gompertz, Weibull, logistic, polynomial and age-period-cohort equations a.o. All these equations result in highly significant correlations between ln mortality rates and age in the age range 35-84 years. This applies to all developed countries and is independent of the differences in causes of death between populations. The best fit is obtained by a second-degree polynomial equation (R2 > 0.99), closely followed by the Gompertz equation. This equation is preferred in view of its extreme simplicity. A highly significant correlation exists between the intercept and the slope of the Gompertz equations, pointing to a crossing-over age. Beyond that age, around 85 years, populations with high mortality rates have a lower mortality, due to selective survival of the strongest individuals. The polynomial age 2 term may be positive or negative, an expression of the acceleration or de-acceleration of mortality at higher ages and is significantly more often positive in women. The equations used are very useful for the study of the aging process and for examining the relationship between possible causal factors and mortality rates in populations. PMID- 12908716 TI - Years of life lost due to premature mortality in Italy. AB - BACKGROUND: The assessment of the burden of disease (BOD) is necessary for sensibly allocating limited health-related resources. No such assessment is available currently for Italy. METHODS: Global burden of disease (GBD) methods were used to analyse in detail the years of life lost (YLLs) component of BOD for the most important diseases contributing to premature mortality in Italy in 1998. YLLs were computed with and without age-weighting and discounting. YLLs were also analysed by gender, for Italy vs. the Euro-A region (a group of 26 European countries) defined in the 2000 update of the GBD Study, and for northern, central and southern Italy, the three traditionally demarcated regions of the country. RESULTS: The use of YLLs yields a ranking of diseases by their relative contribution to mortality burden which differs from a ranking based purely on death counts. Although males contributed 58.5% and females 41.5% of the total mortality burden in terms of YLLs, using death counts the percentages for males and females were similar (50.6% M, 49.4% F). The leading cause of mortality burden, both in terms of YLLs and death counts, was ischaemic heart disease, followed by stroke and lung cancer. Several other conditions, however, had rankings that varied depending on the measure used. While cardiovascular diseases accounted for 31.7% and 'all cancers' for 34.1% of YLLs, they were responsible, respectively, for 44.7 and 27.9% of death counts. The results for Italy generally corresponded with those obtained in GBD 2000 for EURO-A, but the proportion of the total mortality burden explained by the four leading causes is higher in Italy. For within Italy comparisons, there was a decreasing trend from north to south for 'all cancers' and for several specific cancers. Conversely, a consistent increase in YLL rates from north to south was observed for stroke and hypertensive disease. CONCLUSIONS: This analysis of Italy's mortality burden represents the first phase in identifying Italy's total BOD. Simply establishing death counts and rates is no longer sufficient for a full understanding of a country's health status. PMID- 12908717 TI - Serum uric acid and risk for development of hypertension and impaired fasting glucose or Type II diabetes in Japanese male office workers. AB - We examined the association of serum uric acid (SUA) with development of hypertension (blood pressure > or = 140/90 mmHg and/or medication for hypertension) and impaired fasting glucose (IFG) (a fasting plasma glucose level 6.1-6.9 mmol/l) or Type II (non-insulin-dependent) diabetes (a fasting plasma glucose level > or = 7.0 mmol/l and/or medication for diabetes) over a 6-year follow-up among 2310 Japanese male office workers aged 35-59 years who did not have hypertension, IFG, Type II diabetes, or past history of cardiovascular disease at study entry. After controlling for potential predictors of hypertension and diabetes, the relative risk for hypertension compared with quintile 1 of SUA level was 1.27 [95% confidence interval (CI): 1.00-1.62] for quintile 2, 1.34 (95% CI: 1.08-1.74) for quintile 3, 1.48 (95% CI: 1.18-1.89) for quintile 4, and 1.58 (95% CI: 1.26-1.99) for quintile 5 (p for trend <0.001). The respective multivariate-adjusted relative risks for IFG or Type II diabetes compared with quintile 1 of SUA level were 1.55 (95% CI: 0.95-2.63), 1.62 (95% CI: 0.98-2.67), 1.61 (95% CI: 1.01-2.58), and 1.78 (95% CI: 1.11-2.85) (p for trend = 0.030). The association between SUA level and risk for hypertension and IFG or Type II diabetes was stronger among men with a body mass index (BMI) <24.2 kg/m2 than among men with a BMI > or = 24.2 kg/m2, although the absolute risk was greater in more obese men. These results indicate that SUA level is closely associated with an increased risk for hypertension and IFG or Type II diabetes. PMID- 12908718 TI - Tobacco and alcohol consumption, sedentary lifestyle and overweightness in France: a multilevel analysis of individual and area-level determinants. AB - Using a representative sample of the French population, a multilevel analysis allowed us to examine the effect of the level of consumerism, expressed as the gross domestic product (GDP) per capita in the broad area of residence, on the risks of smoking, drinking, sedentary behaviour and overweightness. After adjustment for potential confounders, we found no GDP effect on the odds of being a moderate smoker. Conversely, the risk of being a highly dependent smoker as opposed to a moderate consumer or an abstainer increased with the area-level GDP per capita (OR: 1.13; 95% CI: 1.04-1.23 for an increment of one standard deviation). A similar pattern was found for alcohol consumption: the odds of being a moderate consumer were not related to the GDP per capita, but a positive effect of the GDP on the odds of being an alcohol-dependent drinker as opposed to a moderate consumer or an abstainer was found among women (OR: 1.14; 95% CI: 1.02 1.28). The gap between the sexes with respect to alcohol-dependency therefore appeared to be narrower when the GDP per capita was high. On the other hand, the risk of overweightness was found to increase with the area-level GDP per capita among blue-collar workers only (OR: 1.21; 95% CI: 1.03-1.43). Beyond the well documented socioeconomic effects operating at both the individual and the local neighbourhood levels, our study suggests that broader areas of residence, through their level of economic development, may also have an independent impact on health-related behaviour. PMID- 12908719 TI - Birth weight and risk of angina pectoris: analysis in Swedish twins. AB - OBJECTIVE: Intrauterine nutrition approximated by birth weight has been shown to be inversely associated with risk of coronary heart disease (CHD). By investigating the association within twin pairs discordant for disease, the influence of genetic and early environmental factors is substantially reduced. METHODS: We have investigated the association between birth weight and angina pectoris in same-sexed twins with known zygosity included in the population-based Swedish Twin Registry. Self-reports of birth weight and angina pectoris were collected in a telephone interview between 1998 and 2000. The cohort analyses were based on 4594 same-sexed twins, and the within-pair analyses included 55 dizygotic and 37 monozygotic twin pairs discordant for angina pectoris. Odds ratios (OR) and 95% confidence intervals (CI) were calculated by logistic regression. RESULTS: Compared with birth weight between 2.0 and 2.9 kg, low birth weight (<2.0 kg) was associated with increased risk of angina pectoris in the twin cohort, (OR: 1.46; 95% CI: 1.14-1.87), but after adjustment for potential confounders the risk decreased, and did not reach significance. Within twin pairs discordant for angina pectoris, low birth weight was significantly associated with increased risk of angina pectoris within dizygotic twins (adjusted OR: 5.73; 95% CI: 1.59-20.67), but not within monozygotic twins (adjusted OR: 1.20; 95% CI: 0.40-3.58). CONCLUSIONS: The results suggest that genetic differences associated with foetal growth and adult risk of CHD may have affected previously reported associations between birth weight and CHD. PMID- 12908720 TI - Evaluating a campaign to detect early stage breast tumors in the United States. AB - Growing concern over failure to detect early stage breast cancer has led communities across the United States to participate in the Breast Cancer Awareness Month program. This program mobilizes local public and private institutions, particularly the media, to reach a large audience each October with information on salutary behaviors, including screening, and on resources that can assist newly motivated audiences to adopt the behaviors. Although the scholarly literature includes no assessments of the effect of the program on the actual detection of early stage breast tumors, similar programs targeting other illnesses (e.g., AIDS) are quickly emerging. We attempt such an assessment by applying time-series designs to 92 quarters (beginning January, 1975) of data obtained from cancer registries in the Atlanta and Detroit metropolitan areas as well as in the San Francisco Bay Area. We find that the detection of in situ and local breast tumors increased in all three communities during the quarters that included Breast Cancer Awareness Month. We conclude that community mobilization may have its intended benefit but suggest that community organizers not ignore unintended costs including the emotional and physical sequelae of false positives. PMID- 12908721 TI - Prevalence of hepatitis B and C virus infection in the general population and selected groups in South-Western Greece. AB - Greece is a country of intermediate endemicity for hepatitis B and low endemecity for hepatitis C with a downward trend during the last years. In the present study we investigated the prevalence of hepatitis B virus (HBV) and hepatitis C virus (HCV) infection in the region of South-Western Greece and tried to identify the most important risk factors of transmission. This is a unique epidemiological study, as it is the first community based study in the general population of Greece, with a methodological approach based on multi-staged random sampling. The prevalence of HBV infection seems to be decreasing with a 22.6% rate of HBV markers and a 2.1% rate of chronic HBV carriers. We found male sex, old age and intrafamiliar exposure as the major independent risk factors of HBV transmission, while sexual contact, absence of condom prophylaxis and living in rural areas seem to have also a significant impact for HBV infection. No relation was found between HBV transmission and working in health care facilities, pre-existing hospital admissions and history of transfusion. The prevalence of anti-bodies to the HCV was found 0.5%, even lower than the rate reported in the Mediterranean region. Parenteral exposure was the main risk factor for the transmission of HCV infection. PMID- 12908722 TI - Search for hepatitis delta virus (HDV) infection in hepatitis C patients in north eastern Poland. Comparison with anti-HDV prevalence in chronic hepatitis B. AB - Hepatitis C virus and hepatitis D virus have been shown to suppress HBsAg synthesis. Thus it is possible that HDV infection occurs despite the lack of detectable HBsAg. The aim of our study was to (a) determine the prevalence of HDV infection in patients with chronic hepatitis C (b) compare it with the prevalence of HDV infection in HBsAg positive patients with hepatitis B. The study group consisted of 51 chronic hepatitis C patients, 30 HIV infected drug addicts (27 of them were also positive for anti-HCV) and 102 hepatitis B patients. The participants were tested for anti-HDV, anti-HCV and HBsAg. All anti-HCV positive patients were negative for anti-HDV. Four individuals with anti-HDV belonged to hepatitis B group and constituted 3.9% of all HBsAg positive subjects. We conclude that (a) there is currently no evidence of HDV infection among HCV infected patients in our region (b) hepatitis delta infection is rare in north eastern Poland. PMID- 12908723 TI - Hepatitis C virus infection in injection drug users in Bavaria: risk factors for seropositivity. AB - BACKGROUND: Hepatitis C Virus (HCV) infection is the most common disease among intravenous drug users (IDUs). PATIENTS AND METHOD: All patients admitted to the detoxification unit 1991-1997 and meeting ICD-10 diagnosis of opioid dependency were tested for anti-HCV serology. RESULTS: Thousand and forty nine patients were included in the study. About 61.3% of the IDUs were anti-HCV positive. Increasing age (PR: 1.46; 95% CI: 1.34-1.60), living with a significant other drug user (PR: 1.17; 95% CI: 1.05-1.31), history of therapy (PR: 1.62; 95% CI: 1.50-1.74), history of imprisonment (PR: 1.48; 95% CI: 1.36-1.61), history of emergency treatment (PR: 1.23; 95% CI: 1.12-1.35), additional daily consumption of benzodiazepines (PR: 1.10; 95% CI: 1.00-2.21) or alcohol (PR: 1.26; 95% CI: 1.14 1.38), frequency of injecting heroin (daily: PR: 0.86; 95% CI: 0.78-0.96; previously: PR: 1.14; 95% CI: 1.03-1.26) and type of opioid dependency (methadone: PR: 1.26; 95% CI: 1.13-1.41) were significant factors, considered as individual factors, for positive anti-HCV serology. Using multiple logistic regression we found that older age (OR: 3.54, 95% CI: 1.30-9.67), longer duration of opioid use (OR: 5.74; 95% CI: 1.82-18.13), living with a significant other drug user (OR: 1.47; 95% CI: 1.01-2.16), history of therapy (OR: 4.87; 95% CI: 1.67-14.20), history of imprisonment (OR: 1.92; 95% CI: 1.12-3.28), history of emergency treatment (OR: 1.45; 95% CI: 1.06-1.99) and additional daily consumption of alcohol (OR: 1.49; 95% CI: 1.04-2.13) remained independently associated with positive anti-HCV serology. CONCLUSIONS: These data support the need for early prevention strategies, namely, education of teachers in schools and further training of counsellors informing IDUs of what they can do to minimise the risk of becoming infected or of transmitting infectious agents to others. PMID- 12908724 TI - Comparison of the effectiveness of two mumps vaccines during an outbreak in Switzerland in 1999 and 2000: a case-cohort study. AB - In two recent nation-wide outbreaks of mumps in Switzerland two-thirds of young children with clinical mumps had a history of primary vaccination. On average, measles-mumps-rubella (MMR) vaccination coverage is 80%. Two types of vaccine are commonly used: Jeryl-Lynn and Rubini. The effectiveness of the latter has been questioned in several publications. The authors therefore compared Rubini to Jeryl-Lynn in a case-cohort study. The study included 111 young children with clinical mumps who had been reported to the Swiss Federal Office of Public Health (SFOPH) by primary care physicians of the Swiss Sentinel Surveillance Network (SSSN) between January 1999 and May 2000. Sentinel physicians also sampled 661 children from the same birth cohort as the cases. While we found no evidence for the effectiveness of the Rubini strain, vaccination with the Jeryl-Lynn strain was 70% effective against clinical mumps. Furthermore, children vaccinated with the Rubini strain attended primary health care more frequently with clinical mumps than those who had received Jeryl-Lynn (odds ratio: 2.4; 95% confidence interval (CI): 1.3, 4.7). Restricting the analysis to laboratory confirmed cases increased the odds ratio to 18.4 (95% CI: 2.5, 811.2). Our study confirms the low effectiveness of the Rubini strain vaccine in the field. This vaccine should therefore be considered inappropriate for the control and elimination of mumps and its use should be discontinued. As other vaccines with comparable quality and safety standards and a substantially higher effectiveness are available the MMR vaccination program in Switzerland will not be compromised if the use of Rubini is no longer recommended. PMID- 12908725 TI - Malaria in Turkey: a review of 33 cases. AB - AIM: To evaluate epidemiologic and clinical features of the patients with malaria followed in our clinic, and to review current status of malaria in our country. PATIENTS AND METHODS: Epidemiologic, clinical, diagnostic, and therapeutic features of 33 patients with malaria (4 female, 29 male, mean age: 28 +/- 11 years, range: 15-60) followed in our clinic between 1981 and 2000 were evaluated retrospectively. Malaria data of our country for 1926-2000 were obtained from Health Ministry. RESULTS: Diagnosis was established by thin smears of blood preparations obtained in the febrile period in all cases. Plasmodium vivax was detected in 26 patients (25 domestic and one imported), and P. falciparum in seven (two domestic and five imported). Sixty-one percent of the patients had the prodromal symptoms of the disease and used various antibiotics. All cases demonstrated the typical pattern of fever with chills. Fever (100%), splenomegaly (91%), hepatomegaly (55%), anemia (70%), leukopenia (48%), thrombocytopenia (48%), a rise in sedimentation rate (100%), and abnormalities in hepatic enzymes (30%) were determined in the patients. Chloroquine + primaquine were given to all patients with P. vivax, chloroquin (for three) or mefloquin (for four) alone were given to the patients with P. falciparum. One patient with P. falciparum died soon after admission, all the remaining recovered. Data from Health Ministry revealed that the most common (approximately 100%) species in our country is P. vivax. Although an eradication program against malaria initiated in 1926 achieved success, it still remains as an important health problem. CONCLUSION: Every febrile patient with a history of travel to the regions where malaria is endemic (tropical regions for the world, southeast regions for our country) should raise the suspicion of malaria. Every country should fight against malaria and global cooperation is essential. PMID- 12908726 TI - Seroprevalence of HTLV-I and HTLV-II infection among immigrants in northern Italy. AB - To assess the prevalence of human T cell leukemia virus type I (HTLV-I) and 2 (HTLV-II) infection and the associated risk factors among immigrants living in Northern Italy, we surveyed 3017 open-population subjects from three geographical areas and 371 prisoners. In the open population, the overall prevalence was 0.3% for HTLV-I and 0.1% for HTLV-II, while among prisoners, HTLV-I and HTLV-II infection were detected in 1.4 and 0.8% of subjects, respectively. HTLV-I prevalence was higher in subjects with multiple sexual partners or sexually transmitted diseases. This association was significant in the open-population group and close to significance in prisoners. Multivariate analysis showed that human immunodeficiency virus (HIV) seropositivity remained significantly associated with HTLV-I infection in both targeted populations (OR: 11.2 in the open population; OR: 9.9 among prisoners), whereas sexual exposure was associated with HTLV-I seropositivity only for prisoners (OR: 14.3). No independent variable was related to HTLV-II infection. PMID- 12908727 TI - No incident cases of Chlamydia trachomatis infection in Spanish students after two years of follow-up. PMID- 12908728 TI - Experimental observations and clinical implications of fasting and diet supplementation in fatty livers. AB - Fatty accumulation per se does not appear to affect liver function; however, interest has recently renewed to fatty liver because of the clinical relevance of non alcoholic steato-hepatitis (NASH) and for the increased risk of post transplant failure in grafted livers with steatosis. Clinical and experimental studies have doubtless demonstrated that oxidative stress ensues in steatotic livers. Mitochondria represent the preferential target of the oxidative injury associated to fatty degeneration and show reduced content of glutathione, higher levels of oxidative products and damages to enzymes involved in the process of ATP synthesis, which become more evident under stressing conditions. Although obese patients with fatty liver are advantaged by weight loss, clinical and experimental observations suggest that fatty livers poorly tolerate excessive food deprivation. These observations represent the rationale for treatment strategies based on the supplementation of antioxidants and energetic substrates rather than solely a diet restriction. This review focuses on data emerging from a series of investigations performed in rats with fatty livers induced by a choline-deficient diet, which resembles human steatosis due to an excessive intake of carbohydrates, and aims to give the cue for the development of therapeutic options able to preserve hepatic function after transplantation of steatotic organs. PMID- 12908729 TI - The smooth muscle and airway hyperresponsiveness. AB - Airway hyperresponsiveness, excessive airway narrowing caused by stimuli that normally elicit limited or no response, is one of the cardinal features of asthma. The length-dependence of smooth muscle contractility has been recognized for decades, and it forms an essential foundation for many aspects of the physiological regulation of airway contractility in vivo. This review summarizes the structural and functional alterations of airway smooth muscle in asthma and chronic obstructive pulmonary disease, that underlie pathophysiological conditions of airway hyperresponsiveness. PMID- 12908730 TI - Pre-anesthesiological assessment in paediatric cataract surgery. AB - The authors underline the importance of accurate pre-anesthesiological assessment in children undergoing cataract. The alteration is frequently related to many genetic, metabolic and infectious pathologies that could interfere in anesthesia management whatever surgery is requested. Some possibly responsible congenital syndromes and related alterations are mentioned, particularly focusing on Down's syndrome. PMID- 12908731 TI - Easy, accurate and reliable screening for SNPs by ion pair/reverse phase HPLC: simultaneous detection of factor V G1691A, prothrombin G20210A and methylenetetrahydrofolate reductase C677T variants. AB - We have set up allele specific PCR in combination with high-performance liquid chromatography as a method to simultaneously detect three SNPs, factor V (G1691A), factor II/prothrombin (G20210A) and methylenetetrahydrofolate reductase (C677T) genes, all suspected to be inherited risk factors for deep vein thrombosis. Allele specific multiplex PCR technique was used to generate PCR products of different sizes which were specific for each genotype analyzed in this study. PCR products were analyzed on the WAVE Nucleic Acid Analysis platform by means of a gradient ion pair/reverse phase chromatography without any further purification. This method allows concurrent genotyping of the PCR products of the three mutations within 8 minutes, clearly reduces working time and costs in a routine laboratory, and can easily be adapted to the analysis of any other SNP or the combination of different other SNPs. The assay established here has been validated on more than a hundred samples and gave correct and highly reproducible intra- and interday results, allowing the application of this easy, accurate and reliable test in routine settings and in clinical trials and epidemiological surveys. PMID- 12908732 TI - Evaluation of a fully automated procalcitonin chemiluminescence immunoassay. AB - We evaluated a new fully automated microparticle immunoassay for procalcitonin (LIAISON BRAHMS PCT) in comparison with a previously established manual chemiluminescence assay from the same manufacturer (LUMItest PCT, BRAHMS AG). Procalcitonin (PCT) is an early and rather specific marker of systemic bacterial infection. In addition, the efficacy of antibiotic therapy can be monitored by sequential analysis of PCT values. This is why rapid and accurate determinations of PCT are urgently required by intensive care units. The aim of this study was to evaluate in a clinical set-up a new fully automated rapid PCT test. Analytical results are compared with results obtained by a previously introduced quantitative manual test. Intra-assay coefficients of variation (CV) were found in the range of 0.94 to 7.1% at concentrations between 0.46 and 97.2 microg/l. Over a time period of 27 days the inter-assay CV was found below 4.0% at concentrations of 1.93 and 14.29 microg/l and 9.9% at 0.40 microg/l. The functional sensitivity at a CV level of 20% was determined as 0.2 microg/l. Linearity could be demonstrated in a concentration range from 0 to 445 microg/l. When serum and plasma with EDTA, citrate or heparin anti-coagulation were analyzed in parallel, no systematic bias was found. A method comparison by regression analysis showed PCT values determined by both tests in very good agreement (r = 0.99). PCT concentrations in apparently healthy subjects (n =101) were below 0.58 microg/l in line with previously published results. Patients with sepsis (n = 43) or with infectious adult respiratory distress syndrome (ARDS) (n = 28) showed median values of 22.2 and 18.9 microg/l, respectively. In a clinical set-up the LIAISON Brahms PCT assay provided rapid and accurate PCT results supporting the early detection of severe sepsis, the differentiation between systemic bacterial infection and other inflammatory diseases, and the monitoring of antibiotic therapy in septic patients. The results of the new LIAISON BRAHMS PCT assay show an excellent concordance with the LUMItest PCT. The clinical information derived from the measurements is well comparable to the results obtained with the LUMItest PCT, too. PMID- 12908733 TI - Determination of the cardiac glycosides digoxin and digitoxin by liquid chromatography combined with isotope-dilution mass spectrometry (LC-IDMS)--a candidate reference measurement procedure. AB - This article describes a method of high analytical sensitivity, reproducibility and trueness for the determination of digoxin and digitoxin in serum or plasma at therapeutic levels using a combination of high-pressure liquid chromatography (HPLC), isotope-dilution mass spectrometry (IDMS) and caesium-adduct formation. A method for threefold deuterium substitution in the glycosides was developed, which could be performed within 24 hours without distillation giving yields > 98% of the theoretical value. Extraction from a serum or plasma matrix was performed using a liquid-phase extraction with ammonium acetate buffer/tertiary butylmethyl ether/ethyl acetate at pH 9.5. The HPLC-separation used a 10 x 2 mm LiChrospher RP-18 5 microm guard column in combination with a 125 x 2 mm main column of the same material and a gradient containing methanol, caesium ions and formic acid. Quantification of digoxin and digitoxin was made with IDMS using deuterated internal standards and the system run in single ion monitoring (SIM) mode. The methods had a lower limit of determination of 0.25 microg/l for digoxin and digitoxin, a trueness between 97.5 and 104% for digoxin and between 98 and 101% for digitoxin, respectively and had a coefficient of variation of less than 3% in the therapeutic range for both glycosides. Maximally 1 ml serum or plasma was needed for the procedure. The method is used to set target values for materials used in external quality assessment surveys (EQAS) run by INSTAND as part of a national EQAS-programme.) PMID- 12908734 TI - Reactivity profiles of autoantibodies to different phospholipids and the phospholipid-binding protein beta2-glycoprotein I in patients with clinical symptoms related to thromboembolic and/or vasculopathic events with or without connective tissue diseases. AB - To study the antigenic and epitope specificities of anti-phospholipid Ab in detail, we investigated 177 patients without (62 with APS-related systemic clinical symptoms, 115 with microangiopathies) and 164 patients with connective tissue diseases (CTD). Ab associated with primary APS (pAPS) seem to show a restricted specificity (phospholipid/beta2-GPI-complexes), whereas those in secondary APS (sAPS) react additionaly with pure beta2-GPI. Simultaneously, beta2 GPI-independent Ab were also frequently present in both conditions (50% of all Ab positive sera). In CTD patients, the reactivity profile "pure beta2-GPI + phospholipid/beta2-GPI-complexes" is significantly associated with clinically manifest sAPS. Comparing cardiolipin and phosphatidylserine as antigenic target, the overall concordance (crossreactivity?) between both assays was lower than expected (52%), being highest in pAPS (87%) and sAPS (65%). Based on these results, a two-step procedure for reliable serological diagnosis of APS could be recommended: Ab-screening using a mix of phospholipids complexed with beta2-GPI (sensitivity > 90% for Ab concentrations above 20 U/ml) followed by an assay allowing the simultaneous detection of all relevant antigenic and epitope specificities. PMID- 12908735 TI - The 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) assay as rapid colorimetric method for determination of antibiotic susceptibility of clinical Mycobacterium tuberculosis isolates in liquid medium. AB - We investigated the usefulness of a colorimetric method based on the reduction of a tetrazolium salt (XTT) for the susceptibility testing of clinical isolates of Mycobacterium tuberculosis to isoniazid, rifampin, rifabutin, ethambutol hydrochloride, ethionamide and streptomycin. The isolates and the ATCC reference strains reported as susceptible according to the agar dilution method approved by the National Committee for Clinical Laboratory Standards were found to be susceptible by the XTT colorimetric assay after times of incubation ranging between three days for rifampin and rifabutin to eight days for isoniazid. In comparison with other colorimetric methods reviewed in this article, the proposed assay is suitable for determining the susceptibility or resistance to most antituberculous drugs and, as a consequence of the water-solubility of the formazan yielded by reduction of XTT, additional steps such as the addition of extraction buffer and further incubation before the spectrophotometric analysis are not needed. The XTT reduction assay is an inexpensive, rapid and reliable screening method for the detection of susceptible, resistant and multidrug resistant strains of M. tuberculosis and is an alternative to the costly performance of molecular or radiometric methods. PMID- 12908736 TI - Methodological aspects of the evaluation of postoperative cancer surveillance. Part I: Validity. AB - This contribution deals with problems of determining the validity and efficacy of diagnostic procedures used in postoperative cancer surveillance. While there is a large number of follow-up studies attempting to estimate sensitivity, specificity, or the predictive values of follow-up tests, the methodological aspects of validation are rarely addressed adequately in these publications, and the estimations of validity are often not well-founded. In the first part of this paper we analyze aspects of validity and validation studies. In particular, the following problems are addressed: a) Validity depends on the context; in particular, it depends on the characteristics of the population, the embedding of the test in a diagnostic strategy, the uncertainty of the test itself, the gold standard, and the criterion for positive test results. b) Global, time independent measures of validity are hardly adequate. In particular, any attempt to incorporate a positive lead-time in the definition of the sensitivity is incompatible with an exhaustive classification matrix and with a meaningful definition of predictive values. c) Published estimations of sensitivity mostly suffer from numerous biases. First, there is work-up bias, which arises if positive and negative test-results are verified in a different way (in fact, this implies the use of two different gold standards). Next, test-review bias may occur unless the determination of the test result and the gold standard are mutually blinded. Finally, in quantitative tests, like tumor markers, there is a risk of resubstitution bias if the sample used for defining the cutoff level is identical to the validation sample. Several practical recommendations are given for designing validation studies. In the second part, problems of the efficacy of follow-up programs in regard to patient survival are discussed. Five necessary conditions must be satisfied in order to make such an effect possible. Taking these aspects into account, it can be shown that the usual arguments advanced in favor of diagnostic monitoring are not stringent. As for the design of studies on the efficacy of cancer surveillance, non-randomized studies are inconclusive given that they suffer from a variety of specific biases. It can be shown that the demand, often found in recent publications, that comparative studies of more versus less intensive monitoring be designed so as to permit the detection of a survival advantage in the subgroup of patients who relapse, is unjustified. In the final sections, an overview over published randomized studies and meta analyses on the efficacy of postoperative monitoring of breast and colorectal cancer is given. PMID- 12908737 TI - Methodological aspects of the evaluation of postoperative cancer surveillance. Part II: Efficacy. PMID- 12908738 TI - Agricultural biotechnology: the road to improved nutrition and increased production? PMID- 12908739 TI - Nutritionally enhanced rice to combat malnutrition disorders of the poor. AB - Major deficiency disorders, including vitamin A deficiency, are especially common in countries in which rice is the staple food. In response to the devastating effects of vitamin A deficiency, which may include blindness and, even death, "Golden Rice" has been developed to deliver this nutrient to those populations who need it most. The case of Golden Rice is used to demonstrate the challenges of radical GMO opposition, consumer acceptance, and regulation of biotechnology derived foods. PMID- 12908740 TI - Biotechnology, the environment, and sustainability. AB - This article reviews the environmental impact of genetically modified crops through discussion of environmental risk assessment and includes several case studies. Influence of European regulatory and political issues is considered. PMID- 12908741 TI - Development and transfer of genetically modified virus-resistant sweet potato for subsistence farmers in Kenya. AB - Up to 80% of the yield of sweet potato, the major root crop in East Africa, can be destroyed by crop diseases such as the sweet potato feathery mottle virus (SPFMV). A sweet potato that is resistant to SPFMV is being developed through the combined efforts of non-governmental agenices, biotechnology groups, and agricultural entities. Even small improvements in crop yield, owing to this new technology, will feed countless people in Africa. PMID- 12908742 TI - Productivity improvements in rice. AB - Rice comprises 23%--an immense proportion-of the calories consumed worldwide. Countries that rely on rice as the main staple often consume up to 60% of their daily calories from this cereal product. During the Green Revolution, the yield potential of rice was greatly improved, which is especially important owing to the rapidly growing population. This article reviews the potential contributions of biotechnology to further rice yield potential. PMID- 12908743 TI - A perspective from the food industry. AB - Biotechnology has the potential to meet the ever-increasing demand for enhanced food production. However, several factors besides scientific and agricultural advances can disrupt the progress of this field. This article reviews the interactions between political, regulatory, international, activist, consumer, and scientific entities. A discussion of the precautionary principle is included. PMID- 12908744 TI - An historical perspective from the Green Revolution to the gene revolution. AB - Since the 1960s conventional crop breeding has increased food production commesurate with the growing population. For agricultural development to continue, the exploitation of greater genetic diversity and modern biotechnology are becoming increasingly important. This article reviews the milestones achieved by the Green Revolution and many of the recent breakthroughs of modern biotechnology. PMID- 12908745 TI - Safety assessment of foods produced through agricultural biotechnology. AB - Often the main criticism of foods derived from biotechnology is concerns about food safety. Whereas most present-day biotechnology-derived foods are approximately 99% similar to their non-biotechnology counterparts, the scientific community must ensure the safety of the novel aspects of these foods. The three phases of safety assessment are discussed and the concept of substantial equivalence is explained. PMID- 12908746 TI - Brain imaging techniques. AB - In the last decade, functional brain imaging techniques ranging from fMRI to PET to EEG have added to understanding the mechanisms involved in cognitive, affective, and motoric processes. More recently, as demonstrated by papers in a variety of neuroscience journals, the hypnotic experience and its modulation of critical human processes such as pain have added hypnotic procedures to the experimental techniques available for the neuroscience community. For this interplay to be manifested, it is critical that readers understand brain imaging procedures and the types of questions they can and cannot answer. In this spirit, the present article is an overview of the major brain imaging techniques available to a researcher interested in hypnosis. PMID- 12908747 TI - Hypnosis phenomenology and the neurobiology of consciousness. AB - Recent developments in the philosophical and neurobiological studies of consciousness provide promising frameworks to investigate the neurobiology of hypnosis. A model of consciousness phenomenology is described to demonstrate that the experiential dimensions characterizing hypnosis (relaxation and mental ease, absorption, orientation and monitoring, and self-agency) reflect basic phenomenal properties of consciousness. Changes in relaxation-mental ease and absorption, produced by standard hypnotic procedures, are further associated with changes in brain activity within structures critically involved in the basic representation of the body-self and the regulation of states of consciousness. The combination of experiential and modern brain imaging methods offers a unique perspective on hypnotic phenomena and provides new observations consistent with the proposition that hypnosis is an altered state of consciousness. PMID- 12908749 TI - Temporal aspects of hypnotic processes. AB - The authors examine the cortical processes underlying the process of hypnosis, especially as related to the temporal appearance of specific waveforms in relation to pain. Nonhypnotic pain research suggests that in terms of temporal processing early EEG components are more sensory in nature, and later components are of a more emotional or evaluative nature. In the present work, the authors report that the influence of hypnosis is less on the EEG components associated with the initial sensory experience itself and more on the later components that carry with them rich cognitive/emotional information. The research reviewed in this paper clearly suggests an inhibitory process for the high susceptible individuals associated with the hypnotic analgesia. PMID- 12908748 TI - Negative and positive visual hypnotic hallucinations: attending inside and out. AB - Hypnotic perceptual alteration affects brain function. Those hypnotic instructions that reduce perception by creating an illusory obstruction to it reduce brain response to perception in the cognate sensory cortex, as measured by event-related potential (ERP) amplitude and regional blood flow (PET). Those hypnotic instructions that affect the subject's reaction to perception activate the anterior attentional system, especially the anterior cingulate cortex in PET studies. Hypnosis involves activation without arousal and may be particularly mediated via dopaminergic pathways. Hypnotic alteration of perception is accompanied by measurable changes in both perceptual and attentional function of those specific regions of the brain that process these activities, modulated by the nature of the specific hypnotic instruction. Positive obstructive hallucinations seem to allow for a hypnotic focus inward, activating the functioning of attentional neural systems and reducing perceptual ones. PMID- 12908750 TI - The fox, the hedgehog, and hypnosis. AB - Isaiah Berlin's contrast between the fox, who "knows many things," and the hedgehog, who "knows one big thing," is the starting point for a consideration of monolithic and pluralistic approaches to hypnosis. PMID- 12908751 TI - Rett syndrome as a minicolumnopathy. AB - OBJECTIVE: Rett syndrome is a progressive neurological disorder affecting primarily females. It is characterized by the early regression of acquired language, cognitive functions, social skills, and purposeful hand function. Patients with Rett syndrome are often misdiagnosed as autistic. Recent reports of minicolumnar abnormalities in the brains of autistic and Asperger's syndrome prompted us to search for similar pathology in Rett syndrome. MATERIAL: The patient population consisted of 5 Rett syndrome patients (mean age = 14.4 +/- 4.0 years) and 17 controls (mean age = 14.6 +/- 9.5 years). Tissue was celloidin embedded, sectioned at 35 um and Nissl stained. Images (100x) were taken from Brodmann's areas 9, 21, and 22 from layer III of the left hemisphere. METHOD: Columnar width measurements for these images were obtained with computerized image analysis using previously published algorithms. Each area was analyzed separately with univariate ANOVA, including diagnosis as a fixed factor and age (linear and quadratic terms), and sex as covariates. RESULTS: Diagnosis dependent effects were statistically significant only in area 21 (p = 0.009) even when taking into account a Bonferroni correction for the multiple comparisons. CONCLUSION: Both the regional nature of the changes as well as differences in mean cell spacing differentiates the abnormal minicolumnar morphometry of Rett syndrome from that of autism. PMID- 12908752 TI - Oligodendroglioma: CD44 as a possible prognostic opportunity. AB - CD44, in its standard form as well in its isoforms, is a cell surface adhesion glycoprotein which occurs in a wide variety of non-neoplastic and neoplastic cells. CD44 has been considered to be implicated in tumoral growth and in metastatic potential. We studied the immunohistochemical expression of CD44 standard in 30 oligodendrogliomas (19 primary lesions and 11 recurrences) in order to verify its possible prognostic role. Twelve primary oligodendrogliomas (63%) and 8 recurrences (73%) were CD44-positive. Three of 9 (33%) primary oligodendrogliomas with a Smith grade A-B and 9 of 10 (90%) primary oligodendrogliomas with a Smith grade C-D were found to be in CD44H-positive (p = 0.020). Three of 9 (33%) primary oligodendrogliomas that had not relapsed and 9 of 10 (90%) successively relapsed primary lesions were found to be CD44H-positive (p = 0.020). Median survival of the patients with a CD44H-positive lesion was 84 months; median survival of the patients with a CD44H-negative lesion was 91 months. We conclude that CD44H could have prognostic value regarding the occurrence of relapses. PMID- 12908753 TI - Histological and immunohistochemical study of a neuroblastoma in a dog. AB - A 13-year-old, male German Shepherd dog was euthanasized for a frontal temporal mass revealed by the MRI. The histological examination showed a proliferation composed of small round undifferentiated cells arranged in sheets or nests and sometimes in pseudorosettes interrupted by hypocellular zones of fibrovascular stroma. Immunohistochemical studies revealed the expression of neuroblastic epitopes. The presented neoplasm has many histological and immunohistochemical features in common with the group of olfactory neuroblastomas reported in man, so it could be classified as primitive neuroectodermal tumor with neuronal differentiation. PMID- 12908754 TI - Microsatellite instability and expression of DNA mismatch repair genes in malignant astrocytic tumors from adult and pediatric patients. AB - Microsatellite instability (MSI) is used as a molecular marker for defective DNA mismatch repair (MMR) genes. We report here alterations of MSI in 15 malignant astrocytomas (WHO grade III) and glioblastomas (GBM; WHO grade IV) of pediatric patients (2 - 21 years) and 12 GBM from adults (44 - 68 years) by comparative analysis of BAT25/BAT26 loci and 10 other microsatellite markers. High-level microsatellite instability (MSI-H) occurred in 4 of the 15 pediatric cases (26.7%) and in 1 of the 12 adult GBM cases (8.3%). Low-level microsatellite instability (MSI-L) was observed in 6 pediatric cases (40%) and 8 adult GBM (66.7%). Unstable BAT-25 locus was found in 1 of the MSI-H pediatric cases. Thus, 2 unstable cases showed no instability of this marker. For BAT-26, such a discordance was even more profound: in 1 of MSI-H cases, we obtained no PCR product and the remaining 3 showed no alterations of this marker. MSH2 (Human MutS, Homologue2) protein was detected in all but 3 pediatric cases (1 highly unstable and 2 low-level unstable) and in all adult cases. MLH1 (Human MutL, Homologue 1) protein was detected in all but 2 pediatric cases (1 highly unstable and 1 low-level unstable). Thus, 2 highly unstable pediatric cases showed no detectable MLH1/MSH2 proteins. Our data support earlier observations that MSI occurs predominantly in malignant astrocytic tumors of young patients, which lends support to the hypothesis of different molecular mechanisms of pediatric brain tumors. Surprisingly, we found no significant correlation between the status of 10 microsatellite markers and that of either BAT25 or BAT26 loci or with the expression of MMR genes. PMID- 12908755 TI - Recurrent and atypical meningiomas--a multiparametric study using Ki67 labelling index, AgNOR and DNA Feulgen staining. AB - OBJECTIVE: Histological analysis has limited value to predict the biological behavior of meningiomas. In this study, we investigated the utility of indicators of cell proliferation in the evaluation of histologically benign meningiomas. MATERIALS AND METHODS: For this purpose, 50 meningothelial meningiomas, 50 atypical meningiomas and 8 primary benign meningiomas with their recurrences were studied. For each case the Ki67 labeling index (LI), DNA ploidy and AgNOR were evaluated and the results quantitatively processed and assessed by computerized image analyzer. RESULTS: The Ki67 labelling showed a low index (11.3%) in typical meningiomas and primary meningiomas (13.6%). In contrast, it was higher in atypical (26.6%) and recurrent meningiomas (28%). Similar results were obtained for the AgNOR granule count which showed that typical and primary meningiomas had mean 1.51 - 1.49, whereas recurring meningiomas and atypical meningiomas had mean values of 1.92 and 1.98, respectively. DNA ploidy revealed in the hyperpolyploid region between 4c - 16c: 7.02% of the nuclei in primary meningiomas, 17.98% of the nuclei in recurring meningiomas and 24.63% of the nuclei in atypical meningiomas. CONCLUSION: Our results suggest that evaluation of cell proliferation using Ki67 LI, DNA ploidy and AgNOr, integrated with standard histopathology, can provide better information for a correct grading of meningiomas. PMID- 12908756 TI - Vein of Galen malformation combined with atrial septal defect in a neonate. AB - An arteriovenous fistula (AVF) is an abnormal connection between an artery and a vein, whereby the interconnecting capillary network is missing. Such a malformation frequently occurs in the deep midline regions of the brain, and the subsequent increased flow into the draining vein of Galen substantially dilates in an aneurysmal manner. Congenital forms of the aneurysmal dilatation of the vein of Galen (AVG) often lead to death in the neonatal period, predominantly due to cardiac failure caused by the increased venous inflow as a consequence of the intracerebral arteriovenous shunting. In the presented case a male baby suffered from a rare combination of a cerebral AVF and an atrial septal defect (ASD). He was born at week 38 of pregnancy and subsequently developed tachydyspnoe. Ultrasound (US) and CT scans revealed a large bilateral AVF with dilated basal venous sinuses, hydrocephalus and brain atrophy. In the heart, severe right ventricular hypertrophy, patent ductus arteriosus and an ASD were detectable by US. Neurosurgical consultation rejected the possibility of an operative treatment due to size and localization of the lesion and the existing irreversible brain damage. The child died because of cardiac failure 6 days after birth. Autopsy examination in the brain demonstrated a large conglomerate of dilated blood vessels predominantly in the midline and left occipital lobe, edema and hydrocephalus. In the heart, the ASD detected by US proved to be an ostium secundum-type lesion. Histologically, the conglomerate of vessels revealed features of an AVF and matched the characteristics of AVG. Consequences of chronic ischemic brain injury were also present, with ferruginated neurons suggesting intrauterine damage caused by a congenital AVF. Based on data in the literature, we assume that the left-to-right shunt due to increased venous influx into the heart caused not only cardiomegaly, but may have also interfered with the normal development of the atrial septum leading to an ASD, contributing to the rapid progression of the cardiac failure. PMID- 12908757 TI - Intracranial giant cell arteritis with fatal middle cerebral artery territory infarct. AB - A 37 year-old man who developed a fatal middle cerebral territory infarct was found at autopsy, to have widespread granulomatous angiitis involving meningeal and intracranial--extracerebral vessels but not intracerebral vessels or other extra-cranial vessels. The findings are unique and overlap with those of granulomatous angiitis of the nervous system (GANS) and classic giant cell arteritis (GCA). A possible precipitant for this devastating illness was a recent Chlamydia infection. The salient clinical and pathologic differences between GANS and GCA of the nervous system are discussed. PMID- 12908758 TI - Distal muscular dystrophy of the Miyoshi type. AB - OBJECTIVE: Miyoshi myopathy is an autosomal recessive muscular dystrophy. It is characterized by distal muscle involvement, especially the gastrocnemius and soleus. The disease starts with weakness and atrophy of the calves. MATERIAL AND METHODS: Here we report on 2 patients, brother and sister, from a Turkish family. Onset of the disease was at the age of 20 and 26 years of age, respectively. In both siblings, there was an early and predominant involvement of the distal muscles of the lower limbs. Creatine kinase activity was elevated 50- to 100-fold above normal values. RESULTS: Electromyography revealed a myopathic pattern. Histology of the biceps muscles indicated some myopathic changes consistent with muscular dystrophy. Occurrence in only these 2 siblings with no other family members was indicative of an autosomal recessive inheritance. CONCLUSIONS: We describe the distinctive clinical features in 2 siblings of a Turkish family with MM as differential diagnosis and histological change. PMID- 12908759 TI - Contingency discriminability, matching, and bias in the concurrent-schedule responding of possums (Trichosurus vulpecula). AB - Six possums (Trichosuruus vulpecula) responded under dependent concurrent variable-interval variable-interval schedules of reinforcement. Over 15 conditions, barley-carob was one reinforcer with the other reinforcer consisting of Coco Pops, coconut, or a barley-carob mixture with 0%, 2%, 4%, or 6% salt added to the barley. The schedules were both variable-interval 40 s. As has been found with other species, behavior on the concurrent schedules was biased by the type of feed, with the 6% salt and the coconut giving the greatest biases towards the barley-carob mixture. The schedules were varied over 17 conditions using the barley-carob mixture alone or the barley-carob mixture versus the mixture with 4% or 6% salt. Both the contingency-discriminability model (Davison & Jenkins, 1985) and the generalized matching law described the data from the three sets of conditions equally well. Both gave similar measures of bias; however, some of the parameter values found with the contingency discriminability model were uninterpretable. Thus, any argument for this model based on the interpretability of the parameter values becomes weak. It is worth retaining the generalized matching law as a descriptor of such data. PMID- 12908760 TI - Accuracy of discrimination, rate of responding, and resistance to change. AB - Pigeons were trained on multiple schedules in which responding on a center key produced matching-to-sample trials according to the same variable-interval 30-s schedules in both components. Matching trials consisted of a vertical or tilted line sample on the center key followed by vertical and tilted comparisons on the side keys. Correct responses to comparison stimuli were reinforced with probability .80 in the rich component and .20 in the lean component. Baseline response rates and matching accuracies generally were higher in the rich component, consistent with previous research. When performance was disrupted by prefeeding, response-independent food during intercomponent intervals, intrusion of a delay between sample and comparison stimuli, or extinction, both response rates and matching accuracies generally decreased. Proportions of baseline response rate were greater in the rich component for all disrupters except delay, which had relatively small and inconsistent effects on response rate. By contrast, delay had large and consistent effects on matching accuracy, and proportions of baseline matching accuracy were greater in the rich component for all four disrupters. The dissociation of response rate and accuracy with delay reflects the localized impact of delay on matching performance. The similarity of the data for response rate and accuracy with prefeeding, response-independent food, and extinction shows that matching performance, like response rate, is more resistant to change in a rich than in a lean component. This result extends resistance to change analyses from the frequency of response emission to the degree of stimulus control, and suggests that the strength of discriminating, like the strength of responding, is positively related to rate of reinforcement. PMID- 12908762 TI - Concurrent schedules: reinforcer magnitude effects. AB - Five pigeons were trained on pairs of concurrent variable-interval schedules in a switching-key procedure. The arranged overall rate of reinforcement was constant in all conditions, and the reinforcer-magnitude ratios obtained from the two alternatives were varied over five levels. Each condition remained in effect for 65 sessions and the last 50 sessions of data from each condition were analyzed. At a molar level of analysis, preference was described well by a version of the generalized matching law, consistent with previous reports. More local analyses showed that recently obtained reinforcers had small measurable effects on current preference, with the most recently obtained reinforcer having a substantially larger effect. Larger reinforcers resulted in larger and longer preference pulses, and a small preference was maintained for the larger-magnitude alternative even after long inter-reinforcer intervals. These results are consistent with the notion that the variables controlling choice have both short- and long-term effects. Moreover, they suggest that control by reinforcer magnitude is exerted in a manner similar to control by reinforcer frequency. Lower sensitivities when reinforcer magnitude is varied are likely to be due to equal frequencies of different sized preference pulses, whereas higher sensitivities when reinforcer rates are varied might result from changes in the frequencies of different sized preference pulses. PMID- 12908761 TI - Quantitative analyses of matching-to-sample performance. AB - Six pigeons performed a simultaneous matching-to-sample (MTS) task involving patterns of dots on a liquid-crystal display. Two samples and two comparisons differed in terms of the density of pixels visible through pecking keys mounted in front of the display. Selections of Comparison 1 after Sample 1, and of Comparison 2 after Sample 2, produced intermittent access to food, and errors always produced a time-out. The disparity between the samples and between the comparisons varied across sets of conditions. The ratio of food deliveries for the two correct responses varied over a wide range within each set of conditions, and one condition arranged extinction for correct responses following Sample 1. The quantitative models proposed by Davison and Tustin (1978), Alsop (1991), and Davison (1991) failed to predict performance in some extreme reinforcer-ratio conditions because comparison choice approached indifference (and strong position biases emerged) when the sample clearly signaled a low (or zero) rate of reinforcement. An alternative conceptualization of the reinforcement contingencies operating in MTS tasks is advanced and was supported by further analyses of the data. This model relates the differential responding between the comparisons following each sample to the differential reinforcement for correct responses following that sample. PMID- 12908763 TI - The effect of signaled reinforcement on rats' fixed-interval responding. AB - Four experiments examined the effect on rats' response rate of presenting a brief (500 ms) stimulus simultaneously with the delivery of food on fixed-interval (FI) schedules. In Experiment 1, reinforcement signals that were spatially diffuse (both tones and lights) elevated rates of responding, but responding was attenuated by localized visual stimuli. The remaining experiments examined the signal-induced potentiation of responding. In Experiment 2, a tone reinforcement signal potentiated response rates on an FI schedule, but attenuated response rates on a variable-interval (VI) schedule. This difference was obtained even though the overall rate of responding was equated on the two schedules before the introduction of the signal. Signal-induced potentiation of responding occurred over a range of FI values employed in Experiment 3. In Experiment 4, presenting a reinforcement signal when high local rates of response had occurred immediately before reinforcement resulted in potentiated rates of responding on an FI schedule. The opposite effect on response rate occurred when the reinforcement signal followed only low local rates of response. These results indicate that a variety of factors influence the effects of a reinforcement signal. They imply, however, that the local rate of response at the time of reinforcement is a key factor in establishing the nature of the signaling effect. PMID- 12908764 TI - An analysis of generalized contextual control of conditional discriminations. AB - This research asked whether performance engendered by contextual control procedures would generalize to novel matching-to-sample stimulus arrangements. Two studies were conducted with young adult participants. In Study 1, participants first were trained to perform the contextually controlled conditional discrimination, X-AB, where the sample-comparison relations A1B1 and A2B2 were reinforced in the presence of contextual stimulus X1, but the relations A1B2 and A2B1 were reinforced in the presence of X2. Then, a new conditional discrimination, CD, was established via an unreinforced-conditional-selection procedure. Next, participants were tested for X-CD contextual control performance. Participants selected the originally established CD relations in the presence of X1, but the opposite relations in the presence of X2. Next, an additional conditional relation, EF, was established. Then, participants received trials consisting of entirely novel contextual stimuli, Z1 and Z2, and EF samples and comparisons. Selections were consistent with contextual control; that is, participants selected the originally established EF relations in the presence of one of the novel contextual Z stimuli, but selected the opposite EF relations in the presence of the other contextual Z stimulus. Study 2 systematically replicated these results with naive participants and demonstrated the necessity of first establishing a conditional discrimination prior to tests for generalized contextual control. The findings are discussed in terms of unreinforced conditional selection, stimulus classes, and new ways in which contextual control performances can emerge. PMID- 12908765 TI - Transfer of specific contextual functions to novel conditional discriminations. AB - Three adolescents and 4 children participated in studies designed to examine contextually controlled conditional discrimination performance. In Study 1, participants selected Comparison B1 in the presence one stimulus (A1) and Comparison B2 in the presence of another stimulus (A2) using a matching-to-sample procedure. Next, contextual stimuli X1 or X2 were presented, such that in the presence of X1, selection of B1 given A1 and selection of B2 given A2 were reinforced; and in the presence of X2, selection of B2 given A1 and selection of B1 given A2 were reinforced. Then, new conditional discriminations were taught with Stimuli E and F. When the contextual Stimuli X1 and X2 were presented, participants selected the same comparisons as previously established in the EF relations in the presence of X1, but the opposite comparison as in the EF relations in the presence of X2. The results then were replicated with new Stimuli G and H. In Study 2, a new conditional discrimination, CD, was taught. Then, four combinations of two-element samples--C1 and D1, C2 and D2, C1 and D2, or C2 and D1--were presented with X1 and X2 as comparisons. Five of 6 participants selected X1 in the presence of C1 and D1 or C2 and D2, and selected X2 in the presence of C1 and D2 or C2 and D1. Finally, in Study 3, two new discriminations IJ and JK were taught. Then, the transitive IK relations were tested with X1 and X2 as contextual stimuli. The 4 participants selected K1 in the presence of I1 and K2 in the presence of I2 when the contextual stimulus was X1--demonstrating class formation--and selected the other comparisons when the contextual stimulus was X2. These results suggest that the contextual control functions of X1 and X2 transferred even to relations that had not been directly taught. These results extend those demonstrating generalized contextual control by showing transfer of functions of the contextual stimuli in transitivity tests and when the former contextual stimuli were presented as comparisons. PMID- 12908767 TI - Foamy virus transactivation and gene expression. AB - An overview of the pattern and mechanisms of spuma or foamy virus (FV) gene expression is presented. FVs are complex retroviruses with respect to their genetic outfit and the elements used to control and regulate expression of the viral genome. The increased insight into transcriptional and posttranscriptional mechanisms has revealed that the FVs are distinct, unconventional retroviruses clearly apart from the orthoretroviruses. Although less characterized than the orthoretroviruses, FVs have several unique features that are important for construction and assembly of FV-based vectors for targeted gene delivery and vaccination purposes. Some of these distinguishing features are directly related to the FV-specific mechanisms of gene expression and include (1) the presence of an internal, functional active second transcription unit for expression of the nonstructural genes, (2) the utilization of a subgenomic, spliced transcript for Pol protein expression, and (3) distinct but not yet understood mechanisms for the nuclear exit of defined transcripts and thus an additional level of posttranscriptional control of gene expression. Finally, the interactions of the viral transactivator not only with both viral promoters but also with regulatory elements controlling the expression of defined cellular genes are an important issue with respect to vector development and the apparent apathogenicity of FVs in their natural hosts. PMID- 12908766 TI - The replication strategy of foamy viruses. AB - The replication strategy of foamy viruses diverges in many aspects from what is commonly accepted as the rules of retroviral replication. Although many questions on the details of the replication pathway are still unanswered, it appears that foamy viruses have adopted a strategy which functionally bridges the retroviral and the hepadnaviral replication pathways. A number of experimental findings in favour of the view that foamy viruses are reverse transcribing DNA viruses which integrate into the host cell genome are discussed. PMID- 12908768 TI - Proteolytic processing of foamy virus Gag and Pol proteins. AB - The foamy viral proteases (FV PRs) are set apart from other retroviral processing enzymes by unique features. The first remarkable property is that FV PRs are enzymatically active as high-molecular-mass Pro-Pol proteins. Hence there exist multiple forms of active FV PRs that likely contribute to cleavage site specificity. A FV PR of low molecular size is not detectable in purified virions, in contrast to PRs of other retroviruses that are found in virus particles. Because the major part of Pol remains attached to the amino-terminal enzymatically active PR protein region, the FV-specific way of expressing Pro-Pol polyproteins from a pol-specific transcript provides for the incorporation of Pro Pol and IN into virus particles. Proteolytic processing of Gag and Pol proteins is incomplete and delayed. Another novel feature is that the catalytic center of the active dimers of cat FV PR consists of D-S/T-Q instead of D-S/T-G, an unprecedented feature of this enzyme. The temporal and spatial control and the factors that regulate FV PRs remain to be elucidated. PMID- 12908769 TI - Particle assembly and genome packaging. AB - Foamy virus (FV) replication is distinct from that of all other retroviruses in many respects, including viral assembly. In fact, the viral assembly pathway is rather similar to that of hepadnaviruses such as hepatitis B virus. Foamy virus Gag does not contain landmark retroviral assembly domains such as the major homology region, Cys-His boxes, or a defined M domain. Like hepadnaviruses, the FV Gag protein is not cleaved and contains arginine-rich regions at the carboxyl terminus. In addition, egress of FV particles requires presence of the envelope glycoproteins. Finally, the cis-acting sequences in the FV genome required for genome incorporation, although poorly defined, differ in location from other retroviruses. PMID- 12908770 TI - The foamy virus envelope glycoproteins. AB - The main functions of retroviral glycoproteins are recognition and binding to the cellular virus receptor as well as fusion of viral and cellular lipid membranes to release the viral particle into the cytoplasm of the host cell. Foamy viruses (FVs) are a special group of retroviruses with a very broad host range that use a currently unknown cellular receptor for entry. Nevertheless, many functions of the FV envelope glycoproteins in the viral replication cycle have been characterized in detail over the last years. Several unique features not found for any other retrovirus were identified. These include the presence of two types of FV Env proteins, gp170(Env-Bet) and gp130Env, and the strict requirement of gp130Env coexpression for the FV budding and particle release process, a function that cannot be compensated for by any other viral glycoprotein tested so far. Furthermore, domains in gp130Env could be characterized that influence its intracellular distribution, cell surface transport, and its specific interaction with the viral capsid during particle egress. In addition, it has recently been shown that gp130Env expression alone induces release of subviral particles from cells. This review summarizes the current knowledge about the nature of the FV Env proteins and their function in the viral replication cycle. PMID- 12908771 TI - Foamy virus vectors. AB - Gene therapy is a promising novel treatment for a variety of human diseases. Successful application of gene therapy requires the availability of vehicles with the ability to efficiently deliver and express genes. Viral vectors are efficient means of transferring a gene of interest into target cells. Current available vehicles for gene transfer are either inefficient or potentially unsafe for human gene therapy applications. Foamy viruses offer a fresh alternative vector system for gene transfer with the potential to overcome the concerns of the current vectors. Foamy viruses are nonpathogenic and have a broad host range with the ability to infect various types of cells from different species. Foamy virus replication is distinct and may provide an edge for foamy virus vector usage over other retroviral vectors. These features offer the foamy vectors unique opportunities to deliver several genes into a number of different cell types in vivo safely and efficiently. The principal problems for the design of foamy virus vectors have been solved, and several foamy virus vectors that efficiently transduce a variety of cell types are available. This chapter reviews specific features of foamy virus vector systems and recent advances in the development and use of these vectors. PMID- 12908772 TI - Replication of primate foamy viruses in natural and experimental hosts. AB - Foamy viruses (FVs) are common apathogenic retroviruses readily spread by horizontal transmission in nonhuman primate and some other mammalian host populations. Primate FV infections have been known for half a century, i.e., 15 years before the definition of retroviruses and another 15 years before the detection of primate immune deficiency viruses. The emerging interest in human retroviruses included primate FV, and although the role of human hosts for FV was greatly overestimated temporarily, enthusiastic researchers compiled invaluable data on molecular biology and classic as well as molecular epidemiology of these viruses. It has been shown that lytic FV infection in a wide range of cell cultures is in great contrast to the silent state of the infection in animals. Once transmitted by saliva via biting, FVs reside in all tissues as DNA copies, but their replication is untraceable except in oral submucosal cells, which are thought to supply the virus for transmission. FVs have not definitely been associated with any disease, regardless of viral phylogenetic differences. Various primate and nonprimate species have been used for studies on the natural carrier state and primary infection. Experimental infections have mostly proven to be inefficient in primates as well as lower laboratory animals. However, investigation of the immune response in FV-infected animals has only partly explained the control of FV replication in the animal host. Thus, the biological role of FV remains an enigma to be resolved in the future. PMID- 12908773 TI - Human infection with foamy viruses. AB - Virtually all nonhuman primate species investigated thus far including prosimians, New World and Old World monkeys and apes all harbor distinct and species-specific clades of simian foamy virus (SFV). However, evidence supporting the existence of a human-specific foamy virus (FV) is not yet available. Early reports describing widespread infection of healthy and sick humans with FV could not be confirmed. In contrast, all FV infections documented in humans are of zoonotic origin and are identified in persons occupationally exposed to nonhuman primates. The introduction of SFV into humans raises several public health questions regarding disease outcomes and potential for human-to-human transmissibility. The available data from a very limited number of SFV-infected humans suggest that these infections are nonpathogenic and are not easily transmissible. Additional studies are needed to better define the prevalence and natural history of SFV in humans. PMID- 12908774 TI - Non-primate foamy viruses. AB - Foamy viruses (PFVs), also called spumaviruses, are complex retroviruses inducing a characteristic cytopathic effect in cell culture, leading rapidly to cell lysis. These viruses have been isolated mostly in non-human primates, but three non primate PFVs were characterized, namely the bovine foamy virus, the feline foamy virus and more recently the equine foamy virus. In their hosts, PFVs seem to be apathogenic, mirroring an efficient control of virus replication in vivo. Comparing the biology of the different virus isolates will certainly help to unravel the biology of these retroviruses. PMID- 12908775 TI - Bisphosphonates in oncology: physiopathologic bases and clinical activity. AB - Osteoclastic activation is the ultimate way of bone resorption in neoplasia, induced by the combined effects of tumor-secreted humoral factors (especially parathyroid hormone-related peptides) and osteoclastic-osteoblastic interaction. Bisphosphonates inhibit the osteoclast activity and reduce bone resorption and are a valuable supportive measure for bone disease of neoplasms. Experimental models also suggest an activity of bisphosphonates against cancer cells. Controlled studies, especially in advanced breast cancer and multiple myeloma, indicate different effectiveness against the distinct skeletal-related events. Intravenous clodronate and, especially, pamidronate and zoledronate are the first choice drugs for hypercalcemia, and they play a significant role in reducing metastatic bone pain. Their prolonged use delays, without hampering, the progression of bone disease, including the appearance of osteolysis and the occurrence of pathologic fractures. This effect is probably more valuable when bisphosphonates are administered early in the course of the disease. The evidence that adjuvant bisphosphonates improve survival needs to be confirmed in ongoing studies. Although poorly absorbed by the gastrointestinal tract, oral bisphosphonates are effective in preventing and treating cancer-induced osteoporosis in long-living patients with operable breast cancer. At present, there is little hope that newer bisphosphonates are more effective than those currently used. PMID- 12908776 TI - From conventional to stealth liposomes: a new frontier in cancer chemotherapy. AB - Many attempts have been made to achieve good selectivity to targeted tumor cells by preparing specialized carrier agents that are therapeutically profitable for anticancer therapy. Among these, liposomes are the most studied colloidal particles thus far applied in medicine and in particular in antitumor therapy. Although they were first described in the 1960s, only at the beginning of 1990s did the first therapeutic liposomes appear on the market. The first-generation liposomes (conventional liposomes) comprised a liposome-containing amphotericin B, Ambisome (Nexstar, Boulder, CO, USA), used as an antifungal drug, and Myocet (Elan Pharma Int, Princeton, NJ, USA), a doxorubicin-containing liposome, used in clinical trials to treat metastatic breast cancer. The second-generation liposomes ("pure lipid approach") were long-circulating liposomes, such as Daunoxome, a daunorubicin-containing liposome approved in the US and Europe to treat AIDS-related Kaposi's sarcoma. The third-generation liposomes were surface modified liposomes with gangliosides or sialic acid, which can evade the immune system responsible for removing liposomes from circulation. The fourth-generation liposomes, pegylated liposomal doxorubicin, were called "stealth liposomes" because of their ability to evade interception by the immune system, in the same way as the stealth bomber was able to evade radar. Actually, the only stealth liposome on the market is Caelyx/Doxil (Schering-Plough, Madison NJ, USA), used to cure AIDS-related Kaposi's sarcoma, resistant ovarian cancer and metastatic breast cancer. Pegylated liposomal doxorubicin is characterized by a very long circulation half-life, favorable pharmacokinetic behavior and specific accumulation in tumor tissues. These features account for the much lower toxicity shown by Caelyx in comparison to free doxorubicin, in terms of cardiotoxicity, vesicant effects, nausea, vomiting and alopecia. Pegylated liposomal doxorubicin also appeared to be less myelotoxic than doxorubicin. Typical forms of toxicity associated to it are acute infusion reaction, mucositis and palmar plantar erythrodysesthesia, which occur especially at high doses or short dosing intervals. Active and cell targeted liposomes can be obtained by attaching some antigen-directed monoclonal antibodies (Moab or Moab fragments) or small proteins and molecules (folate, epidermal growth factor, transferrin) to the distal end of polyethylene glycol in pegylated liposomal doxorubicin. The most promising therapeutic application of liposomes is as non-viral vector agents in gene therapy, characterized by the use of cationic phospholipids complexed with the negatively charged DNA plasmid. The use of liposome formulations in local regional anticancer therapy is also discussed. Finally, pegylated liposomal doxorubicin containing radionuclides are used in clinical trials as tumor-imaging agents or in positron emission tomography. PMID- 12908777 TI - Italian general practitioners and smoking cessation strategies. AB - AIMS AND BACKGROUND: General practitioners (GPs) are in a key position to intervene with patients who smoke. The cornerstone of a smoking cessation strategy should be the routine provision of brief advice and follow-up in primary care. However, it seems GPs do not often take action against smoking, at least in Italy as shown by previous reports. The survey was planned, in the context of the "GPs Empowerment Project", a collaborative project involving Denmark, France, Greece, Italy and Portugal, under the ENSP contract with the European Commission (Health & Consumer protection Directorate-General), to evaluate the general attitude, knowledge and behavior of GPs regarding smoking cessation methods. METHODS: A total of 729 family doctors, 409 in Northern and 320 in Southern Italy, were interviewed by phone in the period July-October 2000 regarding their personal smoking habits and their approach with patients on the topic. Prevalence rates were computed using the Epi INFO 6.0 software and were presented as the unadjusted percentage prevalence. RESULTS: The percentage of current smokers among GPs included in the survey was 28.3%, with a higher prevalence in the south (33.3%). Most of the GPs believe that it is their duty to give information about smoking cessation (96.8%) and consider giving information about smoking cessation to their patients an important intervention (98.5%), but only about 49% think their patients will accept their advice. Most GPs (87.3%) declared having discussed about tobacco use with their patients during the month preceding the interview, and 83.5% would like to be trained on smoking cessation strategies. CONCLUSIONS: The results show that, in order to implement primary prevention in clinical practice in Italy, it appears essential to reduce the number of GPs who smoke and to improve GP training on smoking cessation procedures. PMID- 12908778 TI - Screening E-cadherin germline mutations in Italian patients with familial diffuse gastric cancer: an analysis in the District of Urbino, Region Marche, Central Italy. AB - AIMS & BACKGROUND: Hereditary diffuse gastric cancer is a recently defined cancer syndrome caused by inactivating, heterozygous germline mutations in the E cadherin gene (CDH1). To date, 16 truncating germline CDH1 mutations have been described in hereditary diffuse gastric cancer families in different ethnic groups, but so far, no investigation has been addressed to Italian patients. In the District of Urbino, Region Marche, Central Italy, gastric cancer is the most common tumor in men and it is the second in women after breast cancer. In this area, we investigated CDH1 mutations in patients who fulfilled the hereditary diffuse gastric cancer criteria. MATERIAL AND METHODS: Consecutive patients with diffuse gastric cancer were considered eligible for the study. After pedigree analysis, patients who met the International Gastric Cancer Linkage Consortium criteria were studied for CDH1 mutations. After blood samples collection and DNA extraction, standard polymerase chain reaction and sequencing techniques were used for CDH1 analysis. RESULTS: In a study population of 98 patients with diffuse gastric cancer, 11 patients (11%) showed familial clustering and 3 of them met the International Gastric Cancer Linkage Consortium criteria for hereditary diffuse gastric cancer. None of the 3 patients showed inactivating germline mutation in CDH1. CONCLUSIONS: According to recent studies, the frequency of CDH1 inactivating germline mutations in patients who fulfil the hereditary diffuse gastric cancer criteria may be lower than that reported in early investigations. The results of the present study in a population of Italian patients seem to confirm these data. It is likely that unidentified mutations in CDH1 or other involved genes contribute to diffuse gastric cancer susceptibility. PMID- 12908779 TI - Transitions and transversions in Ki-ras gene in colorectal cancers in Mexican patients. AB - AIMS & BACKGROUND: An important increase in the incidence of colorectal cancers has been detected in the last 15 years in Mexico. This fact has been attributed to several causes, including the change in diet acquired from industrialized countries. Various groups have studied the mutational pattern of oncogenes, including Ki-ras gene, in colorectal cancers from different human populations. The aim of this work was to study the prevalence of mutations at codons 12, 13 and 61 of the Ki-ras gene in 37 colorectal tumors from Mexican patients and to correlate them with clinical data. METHODS: Point mutations were studied in 37 colorectal cancers at codons 12 and 13 of the Ki-ras gene, using PCR followed by RFLP. We also performed PCR-SSCP to identify mutations at codon 61. We confirmed mutations by sequence analysis in all the altered codons. RESULTS: Our results indicated that 24.3% of the tumors presented mutations at codon 12, 5.4% at codon 13, and 2.7% at codon 61 of the Ki-ras gene. We found that 75% of these mutations were transitions and 25% transversions. The overall results indicated that the frequency of Ki-ras mutations in colorectal cancers in a sample of a Mexican population (Mexico City) was 32.4%, which is similar to that reported in other populations. We did not find a correlation between the Ki-ras mutations and gender, location of the tumor, or Dukes' stage, but survival of the patient without recurrence was statistically significant. CONCLUSIONS: The study of colorectal cancer indicated that in a Mexican population Ki-ras mutations were present in tumors of patients who survived without tumor recurrence. Most of them were transitions in the first and second base of codon 12. PMID- 12908780 TI - Immunomodulation in a treatment program including pre- and post-operative interleukin-2 and chemotherapy for childhood osteosarcoma. AB - AIMS AND BACKGROUND: The treatment applied in our Institution to children with localized osteosarcoma between 1991 and 1999 consisted of four interleukin 2 (IL 2) courses (9 x 10(6) IU/mL/daily x 4), alternated with pre- and post-operative polichemotherapy. The aims of the present study were to quantify the modifications of some immunological parameters induced by IL-2 and to verify whether polychemotherapy could reduce them. An additional aim was to assess whether any correlation between the immune modifications and the clinical outcome could be found. PATIENTS AND METHODS: We evaluated in 18 consecutive patients the following changes, induced in blood by each IL-2 course: number of lymphocyte subpopulations and natural killer (NK) cells, lymphokine activated killer (LAK) and NK activities. RESULTS: Chemotherapy did not influence the modifications of the number of NK and CD4+ cells and of the LAK and NK activities, induced by each of the four courses of IL-2. The magnitudo of the NK activity and the peak of the NK absolute counts significantly correlated with the clinical outcome. CONCLUSIONS: The results show that the use of IL-2 permitted a repeated immune activation despite the intensive chemotherapy. Furthermore, although the limited number of cases precludes any definitive conclusion, the results suggest a possible role of the NK cells in the control of osteosarcoma. PMID- 12908781 TI - Clinical presentation of peritoneal mesothelioma. AB - AIM: To describe the clinical presentation of peritoneal mesothelioma and its impact on survival. METHODS: Data was collected from 51 patients with peritoneal mesothelioma treated at the Washington Cancer Institute. The demographic, clinical and pathologic information were analyzed. RESULTS: Pain was the most common symptom (recorded in 33% of patients); increased abdominal girth occurred in 31%, increased abdominal girth and pain in 5%, and a new onset hernia in 12%. In an additional 14% of patients, a variety of other clinical findings led to the diagnosis. There was a statistically significant difference in survival by gender, weight loss and volume of disease. CONCLUSIONS: Pain was the most common initial presenting symptom, with increased abdominal girth as a second. A more favorable prognosis occurred in women with a small disease volume. PMID- 12908782 TI - Extrapulmonary small cell carcinoma: multimodality treatment results. AB - AIMS AND BACKGROUND: Extrapulmonary small cell carcinoma is a distinct entity that can occur in many sites, and it is pathologically similar to small-cell lung cancer. We report the results of a retrospective study of a multimodality treatment of 16 consecutive patients with a diagnosis of extrapulmonary small cell carcinoma. METHODS: Primary tumor site was prostate in 2, gallbladder in 2, uterine cervix in 2, liver in 2, endometrium in 1, epididymis in 1, colon in 1, larynx in 1, breast in 1, and unknown primary tumor in 3 patients. Patients' ages ranged from 19 to 79 years (median, 62). Nine patients had limited and 7 had extensive disease. Histologically, 14 were pure extrapulmonary small-cell carcinoma and 2 were mixed with squamous-cell carcinoma. RESULTS: Curative surgery was attempted in 8 patients. Seven patients received local-regional adjuvant radiotherapy. All patients, except the one with a breast primary, were treated with chemotherapy (mostly platinum-based regimens). Overall survival for all patients was 41% and 11% at 2 and 5 years, respectively (median survival, 14 months). Median survival for patients with limited disease was 25 months compared to 12 months for patients with extensive disease (P = 0.05). CONCLUSIONS: Treatment results for extrapulmonary small-cell carcinoma are comparable to those of small-cell carcinomas of the lung. Extent of disease is a significant prognostic factor for survival. PMID- 12908783 TI - Nasopharyngeal/nasal type T/NK lymphomas: analysis of 14 cases and review of the literature. AB - AIMS AND BACKGROUND: Lymphoid malignancies expressing CD56 are rare and most occur in the nasal or nasopharyngeal region. They derive from natural killer cells or from a small subset of T cells that have granular cytoplasm containing molecules that mediate cytotoxic activity: TIA-1, granzyme B and perforin. Both types are closely associated with Epstein-Barr virus. METHODS: We report the pathologic, immunophenotypic and molecular findings in 14 cases of nasopharyngeal/nasal type T/NK lymphomas. RESULTS: Clinically, all patients had localized disease and also had symptoms limited to the nose. The neoplastic cells were frequently pleomorphic, and angiocentric growth was common. Combined immunophenotypic and gene rearrangement analyses demonstrated that most of the cases were true NK cell tumors and were either CD56+ and CD3- or CD56+ and CD3+. Immunohistochemical study showed TIA-1 and granzyme B expression in all cases. By in situ hybridization, most of the cases were associated to Epstein-Barr virus, harboring type 1 virus, and polymerase chain reaction amplification across the 30 bp deletion showed high frequency of latent membrane protein-1-deleted variants. CONCLUSIONS: The nasal type T/NK cell lymphoma shows distinctive clinicopathologic, immunophenotypic and molecular features. These results confirm the important role of Epstein-Barr virus as a local factor in their pathogenesis. PMID- 12908784 TI - Liver damage after transarterial chemoembolization without embolizing agent in unresectable hepatocellular carcinoma. AB - AIM AND BACKGROUND: Transarterial chemoembolization represents a therapy for hepatocellular carcinoma, but in cirrhotic patients affected by large or multifocal HCC with poor hepatic functional reserve, the procedure can damage normal parenchyma. We analyzed the effects on hepatic function of a modified chemoembolization consisting of ethiodized oil (Lipiodol Ultra Fluid) and epirubicin without gelatine sponge (C-LIP). METHODS: Of 90 patients with hepatocellular carcinoma observed from January 1995 to December 1999, 16 with a diagnosis of advanced (large or multifocal) disease underwent 19 C-LIP. The 30th post-C-LIP day was considered as a checkpoint of the biochemical parameters for a possible hepatic failure. The value of alpha-fetoprotein and the clinical finding of ascites were also considered after 30 days. RESULTS: Postoperative values of serum aspartate aminotrasferases, as well as alanine aminotransferase, were significantly higher than preoperative values (P = 0.002 and P = 0.019, respectively) (Table 1). In all patients, there was a significant increase in postoperative total serum bilirubin (P = 0.003). Statistical analysis showed a significant finding of postoperative ascites (P = 0.035) and the effectiveness of C-LIP on neoplastic tissue by a decrease of alpha-fetoprotein values at 30 days (P = 0.067). CONCLUSIONS: Transcatether arterial chemoembolization could represent an effective therapy against multifocal or advanced hepatocellular carcinoma, and its effectiveness is probably not decreased by using a modified procedure without embolizing agent (C-LIP). However, even when performing such a safe procedure, the hepatic functional reserve of the individual patient needs to be accurately evaluated in order to avoid liver failure. PMID- 12908785 TI - The nipple-sparing mastectomy: early results of a feasibility study of a new application of perioperative radiotherapy (ELIOT) in the treatment of breast cancer when mastectomy is indicated. AB - BACKGROUND: Breast cancer surgery has become less and less mutilating, however a mastectomy is required in the case of multicentric, large tumors or recurrences after conservative treatment. The removal of the nipple areola complex during the mastectomy dramatically increases the feeling of mutilation. To reduce this negative psychological impact, in cancers located outside of the central area of the breast, we propose a new type of nipple-sparing mastectomy associated with intraoperative electron beam radiotherapy (ELIOT) delivered on the region of the areola. The nipple-sparing mastectomy is performed leaving 5 mm of glandular tissue behind the nipple areola complex to preserve its blood supply. The reconstruction is immediately performed with a prosthesis or an autologous flap. PATIENTS AND METHODS: Twenty-five patients were included in the study; two of them had a bilateral nipple-sparing mastectomy. RESULTS: Pathological examinations demonstrated the presence of 19 infiltrating carcinomas and 8 ductal carcinoma in situ. Two patients had a superficial skin areolar slough followed by spontaneous healing. One necrosis of the areola occurred due to extensive retroareolar dissection. In the early follow-up, the color of the areola was preserved. All patients except one expressed their satisfaction of having kept their areola. CONCLUSIONS: These preliminary results are encouraging but they require further studies to evaluate the long-term results, the local recurrence rate and the psychological impact. PMID- 12908787 TI - Feasibility of intensity-modulated radiation therapy in the treatment of advanced cervical chordoma. AB - AIMS AND BACKGROUND: Postoperative radiation is often given in cases of cervical chordoma because of the high incidence of local recurrence. The tumor mass usually surrounds the spinal cord and infiltrates vertebral bone. A combined technique using protons or electrons to boost the initial photon fields is generally applied. We evaluated the use of dynamic intensity-modulated radiation therapy as an alternative technique for treating advanced cervical chordoma. METHODS AND STUDY DESIGN: A female patient with incomplete resection of a vertebral chordoma surrounding C2-C3 was irradiated with a total dose of 58 Gy (ICRU point) in 2 Gy daily fractions for 29 days between December 2001 and January 2002. Beam arrangement consisted of seven 6 MV non-opposed coplanar fields. Pretreatment quality assurance included checking of the absolute dose at reference points and 2D dose map analysis. Treatment was delivered with a 120 leaf collimator in sliding window mode. To verify the daily setup, portal images at 0 degrees and 90 degrees were compared with the simulation images before treatment delivery (manual matching) and after treatment delivery (automatic anatomy matching). RESULTS AND CONCLUSIONS: The mean dose to the planning target volume (PTV) was 57.6 +/- 2.1 Gy covering 95% of the PTV per 95% isodose. The minimum dose to the PTV (D99) was 53.6 Gy in the overlapping area between the PTV and the spinal cord planning organ at risk volume (PRV). The maximum dose to the spinal cord was 42.2 Gy and to the spinal cord PRV (8 mm margin) 53.7 Gy. The mean dose to the parotid glands was 37.4 Gy (homolateral gland) and 19.5 Gy (contralateral gland). Average deviation in setup was -1.1 +/- 2.5 mm (anterior posterior), 2.4 +/- 1.3 mm (latero-lateral), 0.7 +/- 0.9 mm (craniocaudal) and 0.43 +/- 1 degree (rotation). CONCLUSIONS: In the treatment of chordomas surrounding the spinal cord, intensity-modulated radiotherapy can provide high dose homogeneity and PTV coverage. Frequent digital portal image-based setup control is able to reduce random positioning errors for head and neck cancer patients immobilized with conventional thermoplastic masks. PMID- 12908786 TI - Surgical management of cerebral metastases from non-small cell lung cancer. AB - AIMS AND BACKGROUND: The objective of the study was to assess the efficacy of surgical resection of solitary brain metastasis in patients with non-small-cell lung cancer. METHODS AND STUDY DESIGN: We report a retrospective analysis of 32 patients with single brain metastasis surgically excised at our hospital. All but one patient underwent postoperative whole brain radiation therapy. RESULTS: The median survival of patients was 12.5 months postoperatively (mean, 17 months), and the overall 1-year survival was 53%. Thirteen patients had recurrence of brain metastasis: 6 of 13 underwent reoperation for the recurrent lesion, and 1 of the 6 patients had a third craniotomy. Baseline characteristics, which significantly influenced survival, included age less than 60 years, tumor histology (ie, adenocarcinoma), and treatment of the primary lung cancer. The analysis did not yield any significant differences between treatment modalities. CONCLUSIONS: Our findings correspond well with those reported in the literature and suggest that surgical resection of single brain metastasis in patients with non-small cell lung cancer can improve survival over conservative management. Furthermore, surgical treatment of the primary tumor and the single brain metastasis, combined or not with radiotherapy and chemotherapy, represents an approach that merits further investigation with more patients and a prospective longitudinal design. PMID- 12908788 TI - Assessment of cytogenetic aberrations and comet assay in colorectal adenocarcinomas. AB - AIMS, BACKGROUND AND STUDY DESIGN: Few studies have investigated the karyotypes of colorectal carcinomas with emphasis on the correlation between cytogenetic findings and clinicopathologic features. The aim of our study involving 20 colorectal adenocarcinomas was to determine their genomic alterations at the chromosomal level by correlating the cytogenetic findings with the extent of DNA damage and clinicopathologic parameters and to compare the results with those of healthy controls. RESULTS: Cytogenetic evaluation of patients and controls revealed 10 abnormal karyotypes in patients with adenocarcinomas located in the rectum, sigmoid and rectosigmoid regions. Four had numerical and six had structural abnormalities. CONCLUSIONS: Statistical analysis revealed a significant difference compared with controls (P <0.01). The karyotypes and the extent of DNA damage assessed by the comet assay were also significantly correlated with tumor stage (P <0.01) using the Kruskal-Wallis non-parametric test, while no statistical significance was observed in relation to patient age and smoking. PMID- 12908789 TI - Cisplatin may be a valid alternative approach in ovarian carcinoma with carboplatin hypersensitivity. Report of three cases. AB - Platinum-based therapy is considered the standard treatment for patients with advanced ovarian cancer. Carboplatin has a more favorable toxicity profile than cisplatin; however, hypersensitivity reactions to carboplatin have been reported occasionally. We reviewed 112 cases of ovarian cancer treated with carboplatin at our institute to identify the hypersensitivity reactions to this chemotherapeutic agent. Hypersensitivity reactions were documented in nine cases (8%). No deaths occurred, but the reactions were judged severe in three of the 112 patients (2.6%). In our own experience with patients showing idiosyncrasy to carboplatin we observed successful resolution after retreatment with cisplatin. Since patients who relapse after initial treatment with carboplatin often respond to it a second time, it is important to continue this treatment. PMID- 12908790 TI - Coincidence of thyroid tumor and thyroglossal duct remnants. Review of the literature and presentation of three cases. AB - AIMS AND BACKGROUND: The coincidence of benign or malignant thyroid tumors with thyroglossal duct remnant (TDR) cysts is rare. Although the precise etiology is still unclear, thyroid origin and spread from a primary site have been suggested and this obviously has important implications for the therapeutic approach. Three cases of thyroglossal duct carcinoma are presented and its management is discussed on the basis of the current rationale for treatment of thyroid cancer. The indication for surgery depends on positive findings in the thyroid gland (nodules, FNAB). The aim of this study was to review our experience in the management of papillary thyroid diseases associated with TDR. MATERIALS AND METHODS: The records of three patients with thyroid tumors associated with TDR treated at the Department of ENT and Head and Neck Surgery of the First Medical Faculty UK of Prague between January 1991 and January 2001 were analyzed. We searched for risk factors of thyroid carcinoma: history of ionizing radiation, history of thyroid diseases, age, tumor size, tumor spread and histopathological factors. RESULTS: We used a triple approach consisting of clinical and ultrasound examination and fine-needle aspiration biopsy for preoperative assessment. Our diagnostic and therapeutic procedures included TDR excision (Sistrunk or Schlange procedure) and total thyroidectomy. Although the therapeutic approach could be a matter of discussion, most patients agreed with our suggestion of relatively radical but non-mutilating treatment. Postoperative radiation or radioiodine ablation is considered in cases of TDR carcinoma or thyroid carcinoma associated with TDR. Oncological follow-up included clinical and ultrasound examination three times during the first year, twice in the second year, and once yearly thereafter. Tumor marker evaluation and/or scintigraphy were performed 6, 12 and/or 24 months following surgery. PMID- 12908791 TI - Leiomyosarcoma of the larynx. AB - Leiomyosarcoma involving the larynx is extremely rare and may be difficult to diagnose. Because of the rarity of this tumor, little information exists on its long-term follow-up and optimal management. We present a review of the literature and report on a patient with leiomyosarcoma of the larynx treated with surgery and postoperative irradiation. In addition, the diagnosis and treatment of leiomyosarcoma are discussed. At six months' follow-up the patient showed no signs of local recurrence but had developed metastases to both lungs. PMID- 12908792 TI - Synchronous presentation of an Askin tumor and a plasmacytoma in an adult patient. AB - Askin tumor, or malignant small round cell tumor of the thoracopulmonary region, is an extremely infrequent entity occurring primarily in children and adolescents. Its histopathologic and cytogenetic features suggest that it belongs to the family of Ewing's sarcoma and primitive neuroectodermal tumors. We report the case of a 43-year-old woman affected by an Askin tumor with bone metastases at diagnosis, presenting synchronously with a plasmacytoma. This is the first reported case of the simultaneous occurrence of an Askin tumor and a malignant hemopathy. The progression of the former and the remission of the plasmacytoma during chemotherapy were remarkable, since Askin tumor treatment shares drugs used for the treatment of plasma cell tumors. Given the infrequent presentation of these diseases in a young adult and the coexistence of two neoplasias characterized by typical chromosomal abnormalities, we consider the possibility of a genetic cancer susceptibility in our patient. PMID- 12908793 TI - Small cell carcinoma of the urinary bladder. A clinicopathologic study of five cases. AB - AIMS AND BACKGROUND: Small cell carcinoma of the bladder (SCCB) is a rare entity characterized clinically by an aggressive behavior with a high incidence of systemic metastases. We report the clinicopathologic findings of five cases. METHODS: We reviewed five consecutive patients with SCCB treated at our institute. In each case the following clinical data were recorded: age, sex, presenting symptoms, endoscopically determined location of the tumor, clinical staging, node involvement (if any), site of metastases (if any), treatment, follow-up and outcome. RESULTS: There were four male and one female patients, age range 42 to 68 years, mean 57.6 years. The clinical presentation was not different from conventional transitional cell carcinoma, with hematuria being the most frequent complaint (four cases). Microscopic examination revealed oat cells in three cases and an intermediate variant in one. At the time of diagnosis the tumors were staged as T3bN2M0, T2N2M0, T4N0M0, T3aN0M0, and T2N0M0. Primary therapy consisted of radical cystectomy alone (one case), transurethral resection (TUR) alone (one case), TUR with chemotherapy (two cases), or TUR with chemotherapy and radiotherapy (one case). Four patients died of progressive disease, with survival from the time of diagnosis ranging from 7 to 16 months (mean, 12.2 months). One patient died of myocardial infarction (unrelated to the primary disease) one month after diagnosis. CONCLUSION: Our study indicates that primary small cell carcinoma of the urinary bladder is as aggressive as its pulmonary counterpart and the overall prognosis of this tumor is very poor. PMID- 12908794 TI - Giant desmoid tumor of the abdominal wall associated with familial adenomatous polyposis. AB - In this paper a case of vast desmoid tumor of the abdominal wall associated with familial adenomatous polyposis is reported. Desmoid tumors represent a particular type of fibrous neoplasms with a higher prevalence in females. They are extremely rare in the sporadic form, while they are found in about 10% of patients affected by familial adenomatous polyposis. Despite their benign histology and the absence of metastatic potential, they can be considered as fibrosarcomas with a low level of malignancy because of their locally invasive nature. The treatment of choice is surgical excision, as wide as possible, aimed at preventing recurrences, which are frequent in this tumor type. The usefulness of complementary therapies such as radiotherapy, hormone or chemotherapy, is not entirely clear. PMID- 12908795 TI - Breast cancer metastatic to the choroid in a male patient: case report. AB - Choroidal metastases have been observed in about 8% of patients with metastatic breast cancer, even if their true incidence is likely to be higher, as they are not routinely investigated in the absence of symptoms. Radiotherapy is the treatment of choice for symptom palliation. The prognosis is traditionally poor, with a reported average survival of one year. Here we describe the third case reported in the literature of a metastatic tumor to the choroid from a male breast carcinoma. PMID- 12908796 TI - Hemorrhagic peritoneal carcinomatosis treated with a weekly schedule of carboplatin and paclitaxel. A case report. AB - Ovarian carcinomas are chemosensitive tumors. Chemotherapy plays a pivotal role also in advanced disease, and the response to chemotherapy appears to be predictive of prolonged survival. Only performance status seems to limit therapy administration and affect patient survival. Here we report on a 66-year-old patient with a clinical status heavily compromised by peritoneal carcinomatosis associated with bloody effusion, which required increasingly frequent paracentesis and transfusions. The clinical conditions worsened under carboplatin monochemotherapy. A further attempt with carboplatin and paclitaxel in a weekly schedule resulted in a clinical response in terms of reduced need for paracentesis and blood support and improved performance status. This case confirms that treatment is the only chance to improve clinical status even in patients with very advanced ovarian cancer and an extensive tumor load. In our opinion, the modified schedule adopted in the case presented here may be worthwhile for future phase II studies in a selected patient population. PMID- 12908798 TI - An extreme consequence of splenectomy in dehydrated hereditary stomatocytosis: gradual thrombo-embolic pulmonary hypertension and lung-heart transplantation. AB - Dehydrated hereditary stomatocytosis (DHS) belongs to the heterogeneous class of hemolytic anemias with leaky red cell membranes. Splenectomy is a highly deleterious treatment, because it favors, with virtually no exception, the occurrence of thromboembolic disease. We describe here the extreme case of a patient with DHS and an associated sickle cell trait. Splenectomy was carried out due to a splenic infarction that occurred during an airplane journey. About 12 years later, the patient noticed an exertional dyspnea, which gradually worsened to such a degree that she became severely incapacitated within 5 years. Eventually, the patient developed a cor pulmonale associated with chronic thromboembolic pulmonary hypertension (CTEPH) and successfully underwent a heart lung transplant operation. This case ranks as one of the most severe examples ever recorded of the effect that splenectomy may have in DHS patients. Nonetheless, it represents the first case to receive a heart-lung transplant. PMID- 12908799 TI - Hb Bronte or alpha93(FG5)Val-->Gly: a new unstable variant of the alpha2-globin gene, associated with a mild alpha(+)-thalassemia phenotype. AB - We report a new unstable variant identified in three carriers of a family from East Sicily; it was named Hb Bronte after the place from which the family originated. DNA sequencing from nucleotides -181 to +894 (alpha1) and to +884 (alpha2) revealed a GTG-->GGG substitution at codon 93 of the alpha2-globin gene. The MCV and MCH values were at the lower end of the normal range in the carriers. On cation exchange high performance liquid chromatography (HPLC), the Hb A2 level was apparently increased to around 6%, and a small abnormal peak (0.3-0.4%) was detected after Hb A2. Two abnormal bands were detected by cellulose acetate electrophoresis: a major band (about 3-4%) migrated between Hb A and Hb F; a minor band (<1%) migrated between Hb A2 and carbonic anhydrase. Normal values of Hb A2 were detected by DEAE microchromatography. On reversed phase HPLC the variant chain was not detected, and most likely it was eluted with the alpha chain peak. The isopropanol stability test was very slightly positive in the carriers. Hemolytic symptoms were absent with the exception of indirect bilirubin, which was at high borderline in 2/3 carriers. In biosynthesis in vitro, the specific activity of the alpha chains was much higher than that of the beta-globin chains, and the alpha/beta biosynthetic ratio in the mother and proband was of the beta-thalassemia (thal) type (2.24 and 2.54, respectively). Time course experiments showed that the increase of the 3H-specific activity of the peak containing normal and variant alpha chains was not linear and was much higher than that of beta chains; moreover, the alpha/beta biosynthetic ratio varied during the 2 hours incubation. PMID- 12908800 TI - Molecular characterization of alpha-thalassemia in Pakistan. AB - Common alpha-thalassemia (thal) rearrangements were studied in a normal random population and in six ethnic groups of Pakistan. Analyses of 204 individuals from the normal population revealed the presence of only the -alpha(3.7) allele with an overall frequency of 8.3%. Ethnic differences were statistically significant for Pashtoon vs. Balochi (p < 0.0005) and Pashtoon vs. Sindhi (p < 0.002). Two hundred and eighty-five thalassemia patients were also studied to identify rare alpha-thal alleles. In this group, 24.6% of the patients had one or two alpha genes deleted. Two rare alleles in the Pakistani population, -alpha(4.2) (0.2%) and alphaalphaalpha(anti3.7) (0.9%), were identified in these patients. The alpha(4.2) allele was found only in Sindhis, while alphaalphaalpha(anti3.7) was present in Punjabis, Sindhis and Balochis. Five patients with triplicated alpha genes were homozygous for either the beta+ or the beta(0) genotype. PMID- 12908801 TI - Role of polymorphic sequences 5' to the G(gamma) gene and 5' to the beta gene on the homozygous beta thalassemic phenotype. AB - Sixty-seven homozygous male and female thalassemic patients with different phenotypes, aged between 8 and 33 years, were divided into three groups, according to the severity of their beta-thalassemia (thal) mutations. We investigated whether some co-inherited genetic factors could influence the phenotype. Patients with milder beta-thal defects, homozygotes or compound heterozygotes for the IVS-I-6 (T-->C) or -87 (C-->G) mutations had a milder disease. In addition, determination of the co-inheritance of the -158 (C-->T) G(gamma) polymorphism and the (AT)9T5 repeat motif in the region -540 to -525, 5' to the beta-globin gene, showed that in some patients with severe or mild/severe beta-thal mutations, linked to haplotype III, there was higher Hb F expression. We conclude that in homozygous beta-thal patients, the severity of the mutations is the most important factor influencing the phenotype, but some polymorphisms such as the -158 (C-->T) G(gamma) and (AT)9T5 repeat motif, increasing the Hb F expression and ameliorate the clinical course of the disease. PMID- 12908802 TI - Hb Lusaka [alpha131(H14)Ser-->phe (alpha1)]: a new variant found in a woman heterozygous for Hb S [beta6(A3)G1u-->Val]. PMID- 12908803 TI - Identification of a new hemoglobin variant: Hb St. Joseph's [beta77(EF1)His- >Leu]. PMID- 12908804 TI - Hb Clinico-Madrid II [alpha90(FG2)Lys-->Arg (alpha1)] and Hb El Escorial [alpha96(G3)Val-->Asp (alpha2)]: two new a chain variants found during a neonatal period study. PMID- 12908805 TI - Two new hemoglobin variants with increased oxygen affinity: Hb Nantes [beta34(B16)Val-->Leu] and Hb Vexin [beta116(G18)His-->Leu]. PMID- 12908806 TI - Identification of a new beta-thalassemia nonsense mutation [codon 59 (AAG- >TAG)]. PMID- 12908807 TI - Mutation at -30 (T-->C) of the delta-globin gene in a Taiwanese beta-thalassemia carrier. PMID- 12908808 TI - Changing cocaine use practices: neo-liberalism, HIV-AIDS, and death in an Argentine shantytown. AB - Cocaine consuming patterns are changing among young drug users who live in "The Villa," a shantytown located in Greater Buenos Aires. After years of drug injection dominance, cocaine snorting became the preferred drug consuming practice while deep and fast structural and cultural transformations have been taken place as part of the neoliberal program implemented in Argentina during the 1990s and the final economic default in 2001-2002. In this article, I analyze how drug users understand and explain these changing practices, including the following aspects: deteriorating economic conditions, the transformations of survival strategies, moral codes, social network organization, violence regulating mechanisms, criminal activity, and police repression. Based on an ethnographic study carried out during the last eight months in "The Villa," I suggest that intense and generalized cocaine injection in shantytowns has logistic, organizational, and structural requirements that cocaine snorting does not have. Particularly, I explore two main aspects associated with these changing cocaine consumption practices: the consequences of the many Acquired Immunodeficiency Syndrome (AIDS)-related deaths, which occurred among older drug injectors, and the progressive social fragmentation tied to the extreme economic deprivation, deepened social exclusion, and growing everyday violence. PMID- 12908809 TI - The separation of codeine from nonprescription combination analgesic products. AB - In the United Kingdom number of products containing codeine in combination with nonopioid analgesics are available from pharmacies for purchase without a prescription. These products may be abused and it has been reported that in order to avoid nonopioid toxicity, some users may attempt to separate the components of combination analgesics, by mixing the dosage form in water and passing it through a coffee filtration apparatus (coffee filters). The present research sought to test the ease of separation of products available in the United Kingdom. The results obtained indicated that separation was possible; however, the extent of separation was influenced by the volume of water used and the product chosen. Healthcare professionals must be aware that separation may be attempted by those wishing to abuse codeine; however, significant nonopioid toxicity could still arise in the abuser. PMID- 12908810 TI - Jail-based substance user treatment: an analysis of retention. AB - Many jail inmates have a history of substance use and "abuse"; few, however, receive comprehensive treatment for substance use disorders while in jail. The authors offer a longitudinal reanalysis of data from five jail-based substance user treatment programs. Survival analysis was used to identify client characteristics associated with length of time in treatment. Survival curves for the five programs were compared, indicating which ones retained inmates the longest. Results from a model stratified by jail site revealed that inmates over 25 years of age and those already sentenced had significantly longer treatment stays. The Substance Abuse Intervention Division (SAID) program, a modified therapeutic community in a New York jail, and the Deciding, Educating, Understanding, Counseling, and Evaluation (DEUCE) program, a curriculum-based intervention, had the longest survival curves and were, therefore, most effective at retaining inmates in treatment. PMID- 12908811 TI - Targeting HIV prevention on African American crack and injection drug users. AB - The use of drugs in the African American community, particularly crack cocaine, has been linked to sexual risk-taking behavior, which increases the likelihood that persons will become infected with Human Immuno Virus. In order to more fully understand risk-taking behavior and to target interventions among African American men and women, this study used data collected from 1277 individuals residing in Lexington and Louisville, Kentucky, who were recruited into National Institute on Drug Abuse (NIDA) Cooperative Agreement Project from 1993 to 1998. The study compared African Americans treated for Sexually transmitted diseases (STDs) (n = 292) with African Americans who reported never being treated for a STD (n = 504) with regard to HIV information, awareness, and the number of HIV tests between those with and without STD exposure. Additionally, we examine gender differences among African Americans who have and have not been exposed to STDs on risk behaviors and HIV knowledge, awareness, and testing. It was hypothesized that African American drug users in the STD group would engage in more risk behaviors than those who reported no STDs. Results indicated that individuals in both groups, the STD exposure group and the no STD group, engaged in similar HIV-risky behaviors. However, the STD group used a greater number of different drugs in their lifetime. The STD group reported they were more likely to get HIV and were more frequently tested for HIV. Females with an STD history were more likely to have been in drug user treatment and to perceive themselves as homeless. Both males and females in the STD group were more likely to report involvement in exchanging sex. PMID- 12908812 TI - Definitions of drunkenness. AB - This study examined definitions of drunkenness in a general population using both quantitative and qualitative data. Using data from 1366 adult current drinkers from the 1995 National Alcohol Survey, frequency of drunkenness and number of drinks to feel drunk were compared. Qualitative data from 58 current drinkers were also examined to determine how they define drunkenness. Overall, there is a positive linear relationship between frequency of drunkenness and number of drinks to feel drunk. For women, the mean number of drinks to feel drunk is below five in all age and ethnicity categories. Several themes emerged from the qualitative data to describe drunkenness that were both negative and positive. These results suggest a possible reframing of how we measure at-risk drinking; a "one-size-fits-all" question of five or more drinks may not be the best way to assess risk for women. PMID- 12908813 TI - The association of selected sociodemographic factors and differences in patterns of substance use: a pilot study in selected areas of Northeast India. AB - Emergence of new addictive substances and progressive increase of 'drug abuse" has become a serious problem in many countries. Despite legal restriction and administrative control, the use of illicit drugs (like opium, heroin, proxyvan, etc.) has increased considerably in many parts of Northeast India. A sample of 1831 people (age 10 years and above) were interviewed during 1997-1998 about their drug use habits, if any, and types of substance used, in Meghalaya and upper Assam. Prevalence of substance use was 29.4% tobacco (20.5% chewers and 12.7% smokers), 12.5% alcohol, and 4.9% opium. In Meghalaya, prevalence of tobacco use was high (41.7%), which was mainly due to the large number of female chewers and male smokers. Opium and cannabis users were mainly confined in Assam close to the Arunachal Pradesh border, indicating a regional influence. Wide differences were observed in the prevalence pattern of tobacco and alcohol use across location, sex, age, education, and occupation. An association of sociodemographic factors with substance use was documented. Religion and ethnic group have significant association with tobacco use patterns but are insignificant with regard to alcohol use. Use of multiple substances and use of cannabis as a substitute for opium are the major problems indicating drug dependence. Mean ages for substance use initiation were: 18.5 years for tobacco, 21.8 years for alcohol, and 25.8 years for opium. Not with-standing the study's limitations, these findings are useful for planned prevention efforts and formulation of a specific program considering the importance of regional and sociodemographic factors. PMID- 12908814 TI - Longitudinal treatment effects among cocaine users: a growth curve modeling approach. AB - This study examined longitudinal treatment effects among cocaine users. The study examined a sample of 371 cocaine users screened from arrestees in jails and from patients in sexually transmitted disease clinics and emergency rooms-all in Los Angeles County during 1992-1994. Of the 371 subjects, 121 had never been in treatment and 250 reported a history of participation in drug user treatment (145 subjects' first treatment was for cocaine use and 105 were in treatment for a drug other than cocaine). Data were collected during face-to-face interviews using a natural history interview instrument. We applied a series of growth curve models to investigate treatment effects on cocaine use. For those who had been in treatment for cocaine use, use of cocaine decreased from approximately 70% before treatment to 12% after treatment entry, while no such changes were observed among those who had never been in treatment or those in treatment for other drugs. Relative to nontreated users, cocaine-treated participants showed a greater likelihood of pretreatment use for both initial status (OR = 3.58) and growth rate (OR = 1.05). After treatment entry, cocaine-treated participants as compared to nontreated participants had a lower likelihood of use (OR = 0.27), although their cocaine use after the initial status increased at a greater rate (OR = 1.03). Treated users were five times less likely to use when they were in treatment than when they were out of treatment. Longer treatment retention was related to initially reduced use but not to later rates of change in cocaine use. The study findings support that treatment for cocaine use is effective in reducing cocaine use. Longitudinal models provide opportunities to demonstrate the dynamic relationships between treatment and outcome. PMID- 12908815 TI - Public health interventionists, penny capitalists, or sources of risk?: assessing street syringe sellers in Hartford, Connecticut. AB - Improved access to sterile syringes decreases risks related to blood-borne transmission of HIV and hepatitis among injection drug users (IDUs). While syringe exchange programs and pharmacy sales of over-the-counter syringes have received considerable attention from researchers and interventionists during the past decade, little is known about informal economy street syringe sellers. In Hartford, CT. as well as other regions throughout the United States a large percentage of IDUs utilize street sellers to facilitate their injection activities. A qualitative and ethnographic study was conducted in Hartford between 1999 and 2001 to help elucidate the public health risks and benefits of street syringe sellers. PMID- 12908816 TI - Expedition Inspiration Fund for Breast Cancer Research Meeting 2003. PMID- 12908817 TI - Genetic polymorphisms of platelet adhesive molecules: association with breast cancer risk and clinical presentation. AB - The main platelet adhesive receptors integrin alpha2beta1, integrin alphaIIbbeta3 and glycoprotein (GP) Ibalpha are also expressed in breast carcinoma cells. They play a key role in tumor cell-induced platelet aggregation and in adhesive interactions necessary for tumoral invasion and metastasis. Several polymorphisms affecting these molecules, two in integrin alpha2 (C807T and G1648A). one in integrin beta3 (T1565C) and one in GP Ibalpha (VNTR), influencing their levels, structure, and possibly their function, have been previously described and associated with cardiovascular diseases. In this study, we investigated the association of these polymorphisms with breast cancer risk or clinical presentation. We studied 101 patients with invasive breast cancer. The main prognostic variables were recorded, and genomic PCR analysis of these polymorphisms was performed. A group of 101 control subjects matched on age and sex was studied and compared with patients. No association was found between VNTR (GP Ibalpha) polymorphism and breast cancer risk or presentation. Genotype and allele frequencies of C807T and G1648A polymorphisms of integrin alpha2 were not statistically different in breast cancer patients and controls, although we found an association between the 1648G/G genotype and higher disease stages (III and IV) (p = 0.02). Breast cancer risk was higher in carriers of beta3 integrin T/T genotype (OR = 2.08, 95% CI = 1.04-4.16, p = 0.04). Furthermore, genotype 1565T/T was also associated with axillary nodal metastasis (p = 0.017) and with tumoral diameter greater than 2cm (p = 0.02). Although confirmatory studies are needed, our results suggest that polymorphic genetic variation of integrins expressed in platelets and epithelial breast cells could modify the risk and the biological aggressiveness of breast carcinomas. PMID- 12908818 TI - H-ras dependent estrogenic effects of epidermal growth factor in the estrogen independent breast cancer cell line MDA-MB-231. AB - A crosstalk between cellular estrogen response and receptor tyrosine kinase signaling has been shown in a variety of estrogen receptor (ER)-positive cell lines. We intended to examine the presence of estrogenic growth factor effects in an ER alpha-negative breast cancer cell line. By means of reporter gene assays, we investigated the activation of estrogen response elements (EREs) by epidermal growth factor (EGF) in the estrogen-unresponsive cell line MDA-MB-231. Our results demonstrate the H-ras-dependent activation of EREs after EGF treatment in this estrogen-unresponsive cell line, an effect which was not observed in the ERalpha/beta-positive breast cancer cell line MCF-7. In MDA-MB-231 cells, the transcriptional activity of an ERE-containing promotor was enhanced dose dependently by all tested EGF concentrations. This effect could be blocked by co treatment with the epidermal growth factor receptor (EGFR) inhibitors AG1478 and ZD1839, as well as by co-transfection with a vector coding for a dominant negative H-ras mutant, but not by co-treatment with the pure antiestrogen ICI182,780. Furthermore, expression of constitutively active H-ras was shown to be sufficient to activate EREs in MDA-MB-231 cells. Our results suggest alternative utilization of ERE-mediated gene regulation in an estradiol unresponsive breast cancer cell line in response to an EGF stimulus. This mechanism was shown to be dependent on EGFR and H-ras activity, but independent of the presence of functional ERalpha. PMID- 12908819 TI - Lignans and tamoxifen, alone or in combination, reduce human breast cancer cell adhesion, invasion and migration in vitro. AB - Flaxseed has been shown to reduce the metastasis of estrogen receptor negative (ER-) human breast cancer in nude mice. This study determined whether enterodiol (ED) and enterolactone (EL), metabolites of plant lignans exceptionally rich in flaxseed, and tamoxifen (TAM), alone or in combination, can influence the various steps of metastasis, that is, breast cancer cell adhesion, invasion and migration, of two ER- human breast cancer cell lines, MDA-MB-435 and MDA-MB-231. The inhibition by ED, EL or TAM (1-5 microM) of cell adhesion to Matrigel or extracellular matrices, fibronectin, laminin, and type IV collagen, as well as cell invasion was dose dependent in both cell lines. When ED, EL and TAM were combined at 1 microM, a greater inhibitory effect on cell adhesion and invasion was observed than with either compound alone. ED and EL at doses of 0.1-10 microM reduced cell migration, but TAM had no effect at 0.1 and 1 microM, and exhibited a stimulatory effect at 10 microM. It is concluded that lignans and TAM, alone or in combination, can inhibit the steps involved in the metastasis cascade. Although more investigations are required, the study also suggests that the intake of the lignan-rich flaxseed may not antagonize the effect of TAM in ER- breast cancer cells. PMID- 12908820 TI - Participation and satisfaction with surgical treatment decision-making in breast cancer among Chinese women. AB - PURPOSE: To report Chinese women's preferred and perceived participation in breast cancer treatment decision making (TDM), describe influences on women's participation preference and participation congruence (PC) (correspondence between preferred and actual amount of participation in TDM), and explore subsequent satisfaction with TDM. PATIENTS AND METHODS: Of 172/211 eligible and available Chinese women recently undergoing breast cancer surgery at one of six Hong Kong government hospitals 154 (89.5%) were recruited. Within 12 days after surgery, women provided interview information on preferred and perceived TDM participation, satisfaction with TDM consultation, difficulties in TDM, and medical and demographic information. RESULTS: Half (55%) reported a treatment choice: 33% wanted the choice to be their own, 59% wanted to share and 8% wanted to delegate the decision. Only age predicted participation preference with older women preferring a more passive role. Eighty percent of women participated as much as, 13% more than and 6% less than desired. Adjusted for age, women reporting PC had fewer difficulties in TDM (beta = 0.21, p = 0.009) than women not reporting PC, while over-involved women had more doubts about their choice (beta = -0.23, p = 0.005). PC was associated with being offered a treatment option (chi2 = 15.59, p < 0.001) and surgeons expressing a surgical preference (chi2 = 6.63, p = 0.036). Satisfaction was unrelated to PC. CONCLUSION: Most Chinese women want shared TDM and to know their surgeon's treatment preference. Over-involved women are at greater risk of difficulties and doubts in TDM and under-involved women perceive a lack of time and information to make their decision. PMID- 12908822 TI - Increase of free insulin-like growth factor-1 in normal human breast in vivo late in the menstrual cycle. AB - Prolonged exposure to endogenous and exogenous sex steroids increases the risk of breast cancer but the mechanisms are poorly understood. Increased levels of circulating insulin-like growth factor-1 (IGF-1) and low levels of IGF binding protein are associated with increased risk of breast cancer suggesting that IGF-1 has to be in its free form to be biologically active. Little is known about sex steroid regulation of IGF-1 locally in the breast. In this study microdialysis was used to determine the local levels of free IGF-1 in normal human breast tissue in healthy female volunteers during the menstrual cycle. The results showed that the extracellular levels of free IGF-1 locally in the breast were doubled in the luteal phase, when estradiol and progesterone levels were elevated, compared with the follicular phase. In plasma, free IGF-1 levels also exhibited a cyclic variation but to a less extent. The increased local levels of the free form of IGF-1 may promote proliferation in the breast epithelium. This could be important in sex steroid dependent breast cancer development. PMID- 12908821 TI - Stable transfection and overexpression of metallothionein isoform 3 inhibits the growth of MCF-7 and Hs578T cells but not that of T-47D or MDA-MB-231 cells. AB - The third isoform of metallothionein (MT-3) is overexpressed in some breast cancers and its expression is associated with a poor disease outcome. In the PC-3 prostate cancer cell line, MT-3 expression has been shown to inhibit cell growth and increase drug resistance. The goal of the present study was to determine if MT-3 overexpression would influence the growth of human breast cancer cell lines. To determine this, the coding sequence of the MT-3 gene was stably transfected into two estrogen receptor positive (MCF-7 and T-47D) and two estrogen receptor negative cell lines (Hs578T and MDA-MB-231) having no basal expression of MT-3. Cell growth was determined by counting DAPI-stained nuclei, cadmium resistance by the colony formation assay, MT mRNA expression by RT-PCR, and MT protein by immuno-blot. It was demonstrated that MCF-7 and Hs578T cells that overexpress the MT-3 gene were growth inhibited compared to untransfected cells. In contrast, T 47D and MDA-MB-231 cells that overexpress MT-3 were not growth inhibited. Stable transfection of the MT-1E gene had no effect on the growth of any of the four cell lines. It was also demonstrated that the overexpression of both MT-3 and MT 1E only increased the resistance of MCF-7 cells to Cd(+2). In all instances, stable transfection of the MT-3 or MT-1E gene had no effect on the expression of the other MT isoforms. The study shows that MT-3 can influence the growth of some breast cancer cell lines. PMID- 12908823 TI - Overexpression of cyclin A overrides the effect of p53 alterations in breast cancer patients with long follow-up time. AB - The tumour suppressor gene p53 and its protein controls critical cellular functions in cell cycle regulation as well as in apoptosis. Recently, in an in vitro study on breast cancer cell line MCF-7, the apoptotic function of p53 has been shown to be altered by overexpression of cyclin A. In this study we have demonstrated a similar association in a consecutive series of 166 breast cancer patients operated for invasive breast carcinomas. We detected mutations (exon 5 8) in the tumour tissue from 28 (16.0%) of the patients, and positive immunoreactivity of p53 protein was detected in tumour tissue samples from 32 (18.8%) patients. A statistically significant correlation between TP53 gene mutations and positive immunohistochemistry of p53 protein was observed (p = 0.0038). Mutations of the TP53 gene, as well as positive immunoreactivity to p53, were associated with poor prognosis (mutations p = 0.053, HR = 1.8, 95% CI 0.99 3.4; positive immunoreactivity p = 0.029, HR 1.33, 95% CI 1.0-1.7; mutation and/or positive immunoreactivity p = 0.015, HR 2.1, 95% CI 1.2-3.7) when cyclin A was not included in the survival analysis. However, when cyclin A overexpression was included, alteration of the p53 protein (mutations and/or positive immunoreactivity) lost its statistical power (p = 0.088). In a stratified survival analysis the OR fell from 3.0 (95% CI 1.2-8.3, p = 0.03) in the low expression cyclin A stratum to 1.3 (95% CI 0.42-4.1, p = 0.77) in the overexpression cyclin A stratum. PMID- 12908824 TI - HER-2/neu assessment in primary chemotherapy treated breast carcinoma: no evidence of gene profile changing. AB - HER-2/neu protein expression and gene amplification were analyzed in a series of 85 consecutive breast carcinoma patients entered into an adriamicin/taxol primary chemotherapy trial followed by surgery, 45 of whom underwent pre-treatment fine needle aspirate (FNA). Dual color FISH (fluorescent in situ hybridization) assay revealed high-level HER-2/neu gene amplification in the immunocytochemistry (ICC) indicated 3+ cases and no, low or moderate amplification in the ICC 2+ group, consistent with previous findings in untreated patients series. Results obtained with the ICC assay CB 11 showed higher overall concordance with FISH than did the Herceptest ICC assay, but CB 11 was less accurate than Herceptest in terms of selecting patients suitable for Herceptin treatment, which is currently restricted to ICC 3+/FISH amplified patients. The only ICC 3+ low-level amplified case (non-amplified according the two more stringent criteria applied) was found with the CB 11 assay. Comparison between pre-treatment smears and post-treatment sections by FISH revealed no significant changes in the HER-2/neu gene profile. In the clinical setting these findings point to the usefulness of HER-2/neu assessment in chemotherapy-treated patients, when pre-treatment material is unavailable. PMID- 12908825 TI - The expression of DCC protein in female breast cancer. AB - BACKGROUND: The deleted in colorectal cancer (DCC) gene has been shown to be frequently deleted or its expression reduced or absent in glioblastomas, colorectal, gastro-intestinal, pancreatic and prostatic tumors. In the present study, we investigated the expression of DCC in surgical specimen from 75 patients with primary breast cancer. METHODS: The expression of the DCC, estrogen receptors (ER), and progesterone receptors (PR) was studied in 75 surgical specimens of primary breast cancer using an immunohistochemical method. To evaluate the outcomes of the breast cancer patients, we followed up the patients during minimum of 10 years. RESULTS: Reduced or loss of expression of DCC was identified in 45 out of 75 samples. There were significant differences between cases without metastasis or local recurrences versus these with metastasis or local recurrences (p = 0.006), and between patients alive with no evidence of malignancy versus those with recurrence or dead of disease (p = 0.005). There were no significant differences between the DCC status and age, sex, tumor location, stage, grade, or proportion of patients who received adjuvant therapy. CONCLUSIONS: These findings suggest that a decrease in DCC expression may influence the prognosis of breast carcinoma patients. PMID- 12908826 TI - Inhibition of invasion and metastasis by glypican-3 in a syngeneic breast cancer model. AB - Glypican-3 (GPC3), a proteoglycan bound to the cell membrane through a GPI anchor, is widely expressed in the embryo but down regulated in most adult tissues, with some exceptions as mammary cells. GPC3 is involved in the regulation of cell proliferation and survival in specific cell types. LM3, a murine mammary tumor cell line unable to express GPC3, was stably transfected with the rat GPC3 gene to analyze its role in tumor progression. Upon injection into syngeneic BALB/c mice LM3-GPC3 clones showed less local invasiveness and developed fewer spontaneous and experimental lung metastasis than controls. GPC3 expressing cells were more sensitive to apoptosis induced by serum depletion, exhibited a delay in the first steps of spreading and were less motile than controls. On the other hand, LM3-GPC3 cells were significantly more adherent to FN than control ones. We observed that GPC3 transfectants presented a higher expression of E-cadherin and beta-catenin, molecules whose down regulation has been associated with tumor progression. Exogenous TGF-beta increased MMP-9 activity in both control and GPC3-expressing cells, but did not modulate MMP-2. Contrarily, GPC3 expression prevented the increase of MMP-2 activity induced by IGF-II. Our results suggest that GPC3 has a protective role against mammary cancer progression. PMID- 12908827 TI - Narrative ability in high-functioning children with autism or Asperger's syndrome. AB - This study examines the narrative abilities of 28 high-functioning children with autism or Asperger's Syndrome and 22 typically developing children across two different discourse contexts. As compared with the typically developing children, the high-functioning group performed relatively well in the storybook context but exhibited difficulty imbuing their narratives of personal experience with the more sophisticated characteristics typically employed by the comparison group. Furthermore, children with autism or Asperger's Syndrome demonstrated impairments inferring and building on the underlying causal relationships both within and across story episodes in both narrative contexts. Findings further revealed that the narrative abilities of children with autism or Asperger's Syndrome were associated with performance on measures of emotional understanding, but not theory of mind or verbal IQ. Findings are discussed in relation to the social and emotional underpinnings of narrative discourse. PMID- 12908828 TI - The comprehension of humorous materials by adolescents with high-functioning autism and Asperger's syndrome. AB - This study investigated the ability of adolescents with Asperger's syndrome or high-functioning autism and an age-matched group of typical adolescents to comprehend humorous materials. The analysis of humor focused on picking funny endings for cartoons and jokes. As expected, the adolescents with autism had significantly poorer comprehension of cartoons and jokes. Both groups had more difficulty with the joke than the cartoon task, but when compared with the typical group, the adolescents with autism performed significantly poorer. Examination of the error patterns revealed that subjects with autism had difficulty handling surprise and coherence within humorous narratives. PMID- 12908829 TI - Further evidence of preserved priming and impaired recall in adults with Asperger's syndrome. AB - Two experiments compared incidental (implicit) and intentional (explicit) memory performance in adults with Asperger's syndrome and individually matched controls. Experiment 1 involved perceptual tests using word fragment cues, following study tasks in which the participants either generated the words from contextual cues or read the words alone, with no contextual cues. Experiment 2 involved conceptual tests using paired associate cues, following study tasks in which the paired associates were rated either for their relatedness or for their readability. Performance in both the incidental tests was similar for both groups. Performance in both the intentional tests was also similar for both groups, with one exception. The adults with Asperger's syndrome were more likely to falsely recall words that had not actually been studied. These findings further delimit the nature of memory impairments in adults with Asperger's syndrome, which seem restricted to certain aspects of episodic memory that include the tendency to make more intrusion errors in recall. PMID- 12908830 TI - Use of functional analysis methodology in the evaluation of medication effects. AB - The atypical antipsychotic medication risperidone was evaluated using a double blind, placebo-controlled design in the treatment of destructive behavior in two individuals with autism. Pre-medication functional analyses indicated that destructive behavior was maintained by escape from demands, attention, or access to tangible items. For both individuals, destructive behavior during the demand condition was significantly reduced during the medication phases, whereas destructive behavior continued to occur to obtain tangible items (Reggie) and attention (Sean). In addition, there appeared to be a differential effect of the medication on self-injurious behavior (SIB) versus aggression for Sean. Results of the study demonstrate how functional analysis may provide information on those conditions and behaviors that are most likely to be affected by a specific medication. PMID- 12908831 TI - Maternal judgments of intentionality in young children with autism: the effects of diagnostic information and stereotyped behavior. AB - This study examined the relation among maternal judgment of intentionality and variables relevant to families of children with autism. Thirty-six mothers of children with autism rated segments of home videotape of behavior from very young children later diagnosed with autism. Mothers were randomly assigned to either a diagnostic or a no diagnostic information group. No significant difference was found on overall ratings of intentionality. Maternal stress was not correlated with overall ratings of intentionality for either group. Mothers in the diagnostic information group rated stereotyped behavior as less intentional. Post hoc analyses showed no differences on maternal ratings of intentionality when the child was in a social setting or interacting with an object, but there were significant differences between ratings when the child was alone. The results are discussed in relation to early development and identification issues in autism. PMID- 12908832 TI - Cognitive underpinnings of pretend play in autism. AB - This article examines the cognitive underpinnings of spontaneous and prompted pretend play in 28 young children with autism, 24 children with other developmental disorders, and 26 typical children. The article compares theories that consider either theory of mind (ToM) or executive function (EF) to be causally important deficits in the development of pretend play in autism and important factors in pretend play. Each of these two theories posits a cognitive precursor to pretense, which would need to be present in typical development, and the absence of which could explain pretend play deficits in children with developmental disabilities such as autism. We tested which of these theories better predicts a child's production of pretend play. Children with autism were significantly delayed on pretend play scores. They also had significant deficits in our ToM measure, but not our EF measures. Regression analyses suggested a role for our measure of generativity, one of the EF measures. PMID- 12908833 TI - Face recognition in children with a pervasive developmental disorder not otherwise specified. AB - This study investigates the accuracy and speed of face recognition in children with a Pervasive Developmental Disorder Not Otherwise Specified (PDDNOS; DSM-IV, American Psychiatric Association [APA], 1994). The study includes a clinical group of 26 nonretarded 7- to 10-year-old children with PDDNOS and a control group of 65 normally developing children of the same age. Two computerized reaction time tasks were administered: a face recognition task and a control task designed to measure the recognition of abstract visuospatial patterns. The latter were either easy or difficult to distinguish from a set of alternative patterns. The normally developing children recognized the faces much faster than the hardly distinguishable abstract patterns. The children in the PDDNOS group needed an amount of time to recognize the faces that almost equalled the time they needed to recognize the abstract patterns that were difficult to distinguish. The results suggest that, when processing faces, children with PDDNOS use a strategy that is more attention-demanding and, hence, less automatic or "Gestalt-like" than the one used by the control children. The results are discussed in the light of a theory that explains the development of coherent mental representations. PMID- 12908834 TI - Autism Diagnostic Interview-Revised and the Childhood Autism Rating Scale: convergence and discrepancy in diagnosing autism. AB - The agreement between the Autism Diagnostic Interview-Revised (ADI-R) and the Childhood Autism Rating Scale (CARS) was investigated in the diagnostic assessment of 54 children aged 22-114 months referred for possible autism. The observed agreement between the two systems was 66.7% (Cohen's kappa = .40) when the ADI-R definition for autism was applied (i.e., scores reaching cutoff in three domains on the ADI-R), but increased considerably with less stringent criteria; that is, scores reaching cutoffs in two domains and in one domain on the ADI-R. As predicted, the CARS identified more cases of autism than the ADI-R. Children classified as autistic according to both instruments had significantly lower IQ/DQ and more severe autistic symptomatology than those classified with the CARS only. PMID- 12908836 TI - The HOXA1 A218G polymorphism and autism: lack of association in white and black patients from the South Carolina Autism Project. AB - A recent study has suggested that the A218G polymorphism in the homeobox A1 (HOXA1) gene may influence susceptibility to autism. We have determined the frequencies of the A and G alleles of the HOXA1 A218G polymorphism in both white and black patients from the South Carolina Autism Project (SCAP) and controls. Marked differences were found in allele frequencies between the races, but no deviations from Hardy-Weinberg equilibrium were seen in either white or black SCAP family members. More direct tests, comparing genotype frequencies between probands and controls and tracking transmission of the A versus G alleles to affected offspring, did not support the contention that allele status for the HOXA1 A218G polymorphism influences one's susceptibility to autism. PMID- 12908835 TI - Analysis of WISC-III, Stanford-Binet:IV, and academic achievement test scores in children with autism. AB - Nonverbal IQs were greater than verbal IQs for young children (3-7 years of age) on the Stanford-Binet:IV (n = 53). However, WISC-III verbal and nonverbal IQs were similar for older children, 6-15 years of age (n = 63). Stanford-Binet:IV profiles were generally consistent for the low-IQ (< 80) and high-IQ (> or = 80) groups, with high scores on visual matching tests (Bead Memory and Quantitative Reasoning). The low- and high-WISC-III IQ groups both performed well relative to IQ on tests of lexical knowledge (Similarities, Information, and Vocabulary), but not on language comprehension and social reasoning (Comprehension). The low-IQ group did best on visuo-motor subtests (Object Assembly and Block Design), but the high-IQ group did not. The high-IQ group had significantly low scores on the Digit Span, Arithmetic, Coding, VMI, and WIAT Written Expression tests, suggesting attention and writing weaknesses. PMID- 12908838 TI - Inclusion to the point of dilution. PMID- 12908837 TI - Brief report: clonazepam behavioral side effects with an individual with mental retardation. AB - Behavioral side effects associated with clonazepam may include agitation, aggression, hyperactivity, irritability, property destruction, and temper tantrums. These side effects may be inadvertently confused with other behavioral or psychiatric conditions, especially if exacerbation of existing challenging behavior occurs. This report describes an individual with mental retardation who experienced behavioral exacerbation associated with clonazepam prescribed at 2 mg/day (0.02 mg/kg/day) to treat aggression, self-injurious behavior, property destruction, and screaming, which was measured with a 15-minute partial interval recording measurement method. When clonazepam was reduced and discontinued, these behaviors significantly decreased from 3.1% of intervals (95% confidence band = 1.6% to 4.6%) to 0.1% of intervals (95% confidence band = 0% to 0.1%). Indicators suggesting review by appropriate medical personnel for possible clonazepam behavioral side effects are provided. PMID- 12908839 TI - The life expectancy of children diagnosed with a pervasive developmental disorder. PMID- 12908840 TI - Integer/linear mathematical programming models: a tool for allocating healthcare resources. AB - In today's environment, the demand for efficient healthcare resource allocation is increasing. As new technologies become available, allocation decisions become more complex and tools to assist decision makers in determining efficient allocations of healthcare resources are encouraged. Mathematical programs have multiple properties that are desirable for healthcare decision makers such as the simultaneous consideration of multiple constraints and a built-in sensitivity analysis. These models have been well researched and are considered invaluable in other industries. Mathematical programming has also become increasingly visible in facilitating the allocation of healthcare resources in the health services research sector. However, the use of mathematical programming tools has been limited in economic evaluations of new technologies. Budget allocations, such as formulary, drug development, and pricing decisions may benefit greatly from the use of mathematical programs. As an increasing number of expensive new technologies become available and pressure grows to contain healthcare costs, these tools may help guide a more efficient allocation of resources for technologies under budgetary and other constraints. PMID- 12908842 TI - Defining monetary values for quality of life improvements: an exploratory study. AB - BACKGROUND: QOL improvements, which can in many cases reflect the overall effect of a health intervention, are frequently measured. It would therefore be useful to define the monetary value of QOL improvements in order to subsequently estimate the value of a given intervention on the basis of its impact on QOL. OBJECTIVE: To explore the feasibility of assessing the monetary value of an intervention by translating the improvement in QOL it produces into willingness to pay (WTP) amounts. METHOD: Seven different hypothetical health state descriptions were developed. Five hundred and forty-five subjects randomly selected among the staff of the University of Montreal, Canada were sent a self administered questionnaire asking them to evaluate the QOL corresponding to various hypothetical health states and indicate the amount they would be willing to pay for interventions that would enable them to move from these health states to perfect health. RESULTS: The response rate was 55% (n = 294). A significant correlation was found between the WTP amount and the anticipated QOL improvement, and between the WTP amount and the respondent's family income. A regression equation for WTP amounts was derived from a step-wise model including the anticipated QOL improvement and the respondent income. While the model could only account for about 20% of the WTP variance (R(2) = 0.2), the mean predicted WTP amounts were very close to the mean WTP amounts observed in the sample. CONCLUSION: This study indicates that a definite correlation exists between willingness to pay for an intervention and the improvement in QOL it produces. PMID- 12908841 TI - Recurrent genital herpes treatments and their impact on quality of life. AB - Herpes genitalis is one of the most common viral sexually transmitted diseases in the world, with an estimated seroprevalence in the US of greater than 20%. Two viruses of the same family cause herpes genitalis: herpes simplex virus 1 and 2. After the resolution of primary infection, the virus persists in the nerve roots of the sacral plexus, often causing recurrent (though generally less severe) outbreaks. These outbreaks, as well as the infectious potential to the patient's sexual partners, results in significant psychological stress on the patient, and has a tremendous negative impact on QOL. Current treatment modalities may result in a reduction in the number of outbreaks and viral shedding, but no cure exists. Although studies have clearly demonstrated the negative impact of recurrent genital herpes on QOL, an assessment scale specific to herpes was not developed until recently. Earlier studies indicated that patients did not perceive a significant benefit from episodic treatment with antivirals, but studies using the Recurrent Genital Herpes Quality of Life Questionnaire (RGHQoL) have now demonstrated that suppressive antiviral therapy improves quality of life in patients with frequent recurrences of genital herpes. However, not all patients with recurrent genital herpes need suppressive therapy, and proposed factors to consider include frequency of recurrence, physical and psychological distress caused by recurrences, and the potential for transmission to the patient's sexual partner. Newer therapeutic modalities, including the topical immune response modifier resiquimod and herpes vaccines, may eventually be shown to further decrease the psychological morbidity of recurrent genital herpes. PMID- 12908843 TI - Validation of expert opinion in identifying comorbidities associated with atopic dermatitis/eczema. AB - BACKGROUND: The use of expert opinion is widespread in economic studies of healthcare utilisation; however, few studies have attempted to assess the validity of assumptions derived from such sources. OBJECTIVE: To examine the use of such expert opinion in determining comorbidities associated with atopic dermatitis/eczema (AD/E), which were assessed as part of a recent third-party payer cost-of-illness study. DESIGN: To identify the disease-related comorbidities that would represent costs associated with AD/E, physicians on an expert panel were asked individually and then collectively to group all International Classification of Diseases, 9(th) Edition-Clinical Modification (ICD-9-CM) diagnosis codes as 'most likely', 'possibly' or 'definitely not' related to the costs of identifying and treating patients with AD/E. Claims representing $US464 million in payer reimbursements from nearly 125 000 patients with AD/E were identified within two separate claims databases (1997 values). Over 850 ICD-9-CM diagnosis codes were identified in the first-listed position from these claims. For each group of 'most likely', 'possibly' and 'definitely not' related diagnosis codes, prevalence rates were compared within AD/E and non AD/E populations from the two historical payer claims databases. Adjusted and non adjusted odds ratios were calculated by comparing prevalence rates between AD/E and non-AD/E patients in the same payer population. RESULTS: The mean prevalence rate of any diagnosis code in the AD/E population was 0.65 +/- 1.82% (SD) with a mean odds ratio of 1.81 +/- 0.96. Comorbidities considered by the expert panel 'most likely' to be associated with AD/E had higher prevalence rates (3.28 +/- 3.63%) and odds ratios (2.14 +/- 1.14). Comorbidities considered to be 'possibly' related to AD/E had prevalence rates and odds ratios of 3.01 +/- 5.06% and 1.84 +/- 0.82, respectively. Comorbidities considered to be 'definitely not' related to AD/E had the lowest prevalence rates (0.45 +/- 1.09%) and odds ratios (1.80 +/ 0.97). CONCLUSIONS: Comparing the result of consensus panels with actual claims histories validated the use of expert opinion in determining comorbidities associated with AD/E. Expert opinion yielded valid results in terms of identifying comorbidities that manifested frequently and disproportionately in the AD/E population. Limited statistical measurements of comorbidities would have been less specific than expert opinion. Future cost-of-illness studies should consider alternative data sources and methodologies to enhance the validity and importance of expert opinion and to corroborate their findings. PMID- 12908844 TI - Eptifibatide: a pharmacoeconomic review of its use in percutaneous coronary intervention and acute coronary syndromes. AB - Eptifibatide (Integrilin) is a selective inhibitor of platelet glycoprotein (GP) IIb/IIIa receptors used as adjunctive therapy for patients undergoing percutaneous coronary intervention (PCI) and for patients with acute coronary syndromes (ACS), particularly those requiring PCI. Most economic analyses of eptifibatide have incorporated clinical and healthcare resource use data from either the ESPRIT (Enhanced Suppression of the Platelet IIb/IIIa Receptor with Integrilin Therapy) study in low- to moderate-risk patients undergoing selective PCI with stent implantation or the PURSUIT (Platelet Glycoprotein IIb/IIIa in Unstable Angina: Receptor Suppression Using Integrilin Therapy) trial in patients with ACS. Eptifibatide achieved statistically significant reductions in combined endpoints of death and ischaemic complications in both of these large multicentre clinical trials, in which patients were randomised to receive intravenous eptifibatide or placebo as adjunctive therapy to heparin and aspirin (plus a thienopyridine in ESPRIT). In US economic analyses using ESPRIT trial data, approximately 40% and 70% of the acquisition cost of eptifibatide was offset by reduced medical resource consumption during the initial hospitalisation period and over a 1-year period, respectively. Eptifibatide was associated with a favourable cost-effectiveness ratio of $US1407 (year 2000 costs) per life-year gained (LYG) in a retrospective US cost-effectiveness analysis that incorporated data from the ESPRIT trial and modelled life expectancy using a large cardiovascular database.Several cost-effectiveness analyses used prospectively collected data from the PURSUIT trial and modelled survival projections using similar methods. These analyses, conducted in the US, Canada and Western Europe, also showed favourable results ($US3761-$US18 774 per LYG; various years of costing). Cost-utility ratios reported in US analyses varied somewhat, but remained <$US20 000 per quality-adjusted life-year gained (1996 values) when clinical efficacy data were derived from the US cohort of PURSUIT. CONCLUSION: Significant clinical benefits have been demonstrated with eptifibatide as adjunctive therapy in patients undergoing selective PCI with stent implantation in the ESPRIT trial and in patients with ACS in the PURSUIT trial. Pharmacoeconomic analyses using data from either ESPRIT or PURSUIT have demonstrated favourable cost-effectiveness ratios for both indications in various countries. ESPRIT-based results from the limited number of available economic analyses are particularly favourable. The cost-effectiveness of eptifibatide in ACS (i.e. PURSUIT-based results) may be further improved by targeting the drug for patients in whom catheterisation and PCI are planned, although further analyses are required to confirm this. PMID- 12908845 TI - Tolcapone-related liver dysfunction: implications for use in Parkinson's disease therapy. AB - Levodopa is the cornerstone of idiopathic Parkinson's disease (PD) treatment. However, after long-term use of levodopa, a significant percentage of patients experience motor fluctuations, which worsen their quality of life. Catechol-O methyltransferase (COMT) inhibitors reduce levodopa metabolism and enhance the respective plasma levels, resulting in improvements in symptoms and overall quality of life. Tolcapone was the first drug of this class to be marketed, but was withdrawn in the European Union due to its implication in the deaths of three PD patients due to hepatic failure. Three deaths from fulminant hepatic failure in 40000 patient-years is a number that is 10-100 times higher than the expected incidence in the general population and, according to the manufacturer's own information, the number is probably underestimated due to under-reporting of cases. In the US, tolcapone was not withdrawn, but restrictive liver enzyme monitoring measures were issued by authorities, which severely limited its use. No further deaths from hepatic failure were reported since these measures were implemented. The mechanisms by which tolcapone may induce liver toxicity are still under debate. It was thought that mitochondrial uncoupling of oxidative phosphorylation by tolcapone, and consequent impairment of energy production by hepatocytes, could be responsible for the observed effects. Some experts consider that the restrictive guidelines issued in the US regarding tolcapone use may be loosened with no consequential reductions in safety. It was suggested that ongoing clinical information about safety should be considered and periodical revisions of the restrictions made accordingly. The identification of the molecular and biochemical basis of tolcapone hepatotoxicity, when completed, should also provide important indications for the clinical use of this drug. In conclusion, appropriate monitoring of liver function can ensure adequate safety in PD patients receiving tolcapone, who can therefore benefit from the symptomatic improvements obtained with this drug. PMID- 12908846 TI - Drug-induced glaucomas: mechanism and management. AB - Glaucoma comprises a heterogeneous group of diseases that have in common a characteristic optic neuropathy and visual field defects, for which elevated intraocular pressure is the major risk factor. The level of intraocular pressure within the eye depends on the steady state of formation and drainage of the clear watery fluid, called the aqueous humour, in the anterior chamber of the eye. An obstruction in the circulatory pathway of aqueous humour causes an elevation in intraocular pressure. Because intraocular pressure is the most modifiable parameter, therapeutic measures (medical and surgical) are aimed at reducing the pressure to protect against optic nerve damage. Glaucomatous optic neuropathy results from degeneration of the axonal nerve fibres in the optic nerve and death of their cell bodies, the retinal ganglion cells. Clinical examination of the optic nerve head or disc and the peripapillary nerve fibre layer of the retina reveals specific changes, and the resulting visual field defects can be documented by perimetry. Glaucoma can be classified into four main groups: primary open-angle glaucoma; angle-closure glaucoma; secondary glaucoma; and developmental glaucoma. Drug-induced glaucoma should be considered as a form of secondary glaucoma because it is brought about by specific systemic or topical medications. Although there is a high prevalence of glaucoma worldwide, the incidence of drug-induced glaucoma is uncertain. Drugs that cause or exacerbate open-angle glaucoma are mostly glucocorticoids. Several classes of drugs, including adrenergic agonists, cholinergics, anticholinergics, sulpha-based drugs, selective serotonin reuptake inhibitors, tricyclic and tetracyclic antidepressants, anticoagulants and histamine H(1) and H(2) receptor antagonists, have been reported to induce or precipitate acute angle-closure glaucoma, especially in individuals predisposed with narrow angles of the anterior chamber. In some instances, bilateral involvement and even blindness have occurred. In this article, the mechanism and management of drug-induced glaucomatous disease of the eye are emphasised. Although the product package insert may mention glaucoma as a contraindication or as an adverse effect, the type of glaucoma is usually not specified. Clinicians should be mindful of the possibility of drug induced glaucoma, whether or not it is listed as a contraindication and, if in doubt, consult an ophthalmologist. PMID- 12908847 TI - Benefits and risks of simvastatin in patients with familial hypercholesterolaemia. AB - Familial hypercholesterolaemia is a frequent, inherited, monogenic disorder, associated with accelerated development of atherosclerotic disease leading to coronary artery disease. Life expectancy of patients with familial hypercholesterolaemia is reduced by 15-30 years unless they are adequately treated with lipid-lowering therapy. Given the chronic nature of this disease, the selection of a therapeutic approach should be strongly based on its long-term safety and tolerability. The introduction of HMG-CoA reductase inhibitors has revolutionised the treatment of familial hypercholesterolaemia. Simvastatin 40-80 mg/day effectively reduces serum low density lipoprotein (LDL)-cholesterol levels. Furthermore, simvastatin reduces triglycerides and mildly raises high density lipoprotein-cholesterol levels. In addition to the hypolipidaemic effect, other potentially important effects, such as improvement of endothelial function and reduction of LDL oxidation and vascular inflammation, have been associated with HMG-CoA reductase inhibitor therapy. Simvastatin has also been shown to abolish the progression, and even facilitate the regression, of existing human atherosclerotic lesions. The good safety and tolerability profile of simvastatin is clearly highlighted by the low rate of therapy discontinuation observed in several population-based clinical trials. The most common adverse events leading to the discontinuation of therapy are gastrointestinal upset and headache. Asymptomatic elevations in liver transaminase levels and myopathy are uncommon. The overwhelming clinical evidence regarding the long-term use of HMG-CoA reductase inhibitor therapy in patients with familial hypercholesterolaemia together with the long-term safety data (particularly relating to simvastatin) provide support for the use of this drug as a first-line agent when pharmacological treatment is indicated. Early intervention with simvastatin treatment can be successfully implemented with favourable economic benefits. PMID- 12908848 TI - Safety and pharmacology of oseltamivir in clinical use. AB - Oseltamivir is a novel agent approved for the treatment and prevention of influenza infection and illnesses in adults and children. Assessment of data from the clinical trial programme, a US health insurance database study and postmarketing surveillance allowed a comprehensive review of the safety of oseltamivir in clinical use in subjects >1 year of age. Oseltamivir has been studied over the course of a 5-year development programme in >11000 subjects from North America, Europe and the Southern Hemisphere, including otherwise healthy adults, approximately 500 elderly/high-risk subjects, and children (>1000) aged 1 12 years. Safety evaluations included treatment-emergent adverse events, hospitalisations and deaths, as well as haematological and biochemical laboratory safety tests. The data reveals that oseltamivir has simple, uncomplicated pharmacology and lacks potential for drug-drug interactions. Electrocardiogram parameters, including corrected QT interval, were unaffected by oseltamivir even at high doses. Postmarketing studies confirmed that transient gastrointestinal disturbance is the major adverse effect of oseltamivir and that this can be reduced by taking oseltamivir after a light snack. On treatment serious adverse events were reported in 1.3% of oseltamivir 75mg twice daily, 0.7% of oseltamivir 150 mg twice daily and 1.2% of placebo recipients, respectively, in the clinical trial programme. Postmarketing, it is estimated that, to date, over 4 million oseltamivir prescriptions have been dispensed worldwide. Approximately 2300 spontaneous reports were received by the manufacturer over the three winter seasons of use. As these events are reported infrequently and from an unknown number of users, it is not possible to definitively assess causality or frequency of reported events. Most reports were of gastrointestinal and skin reactions. However, a clear association between the skin reactions and oseltamivir has not been established. A large study of insurance records, which permitted the assessment of the relative risk of medical events treated in the month following prescription of oseltamivir in general use, showed no evidence of increased risk of cardiac, neuropsychiatric or respiratory events for those receiving oseltamivir compared with those who did not. To conclude, no important safety concerns have evolved which might limit the suitability of oseltamivir for the treatment and prevention of influenza in all patient populations. PMID- 12908849 TI - A feasibility study for recording of dispensing errors and near misses' in four UK primary care pharmacies. AB - Medication errors can occur at the prescribing, transcription, dispensing and administration stage of drug therapy. However, publication of the Organisation with a Memory (OWAM) by the Department of Health in the UK has raised awareness of the need for pharmacies to collect information about dispensing errors and near misses, which occur within an organisation. Such information provides valuable insights into the vulnerabilities of dispensing procedures and identifies areas for improvement in dispensing systems The main aim of this study was to investigate the feasibility of a self-reporting system for dispensing errors and near misses in primary care (community) pharmacies. It was also to identify the types of errors or near misses commonly encountered in community pharmacies. A data collection form was designed and modified for use after a pilot study. Four community pharmacies volunteered to participate in this feasibility study. The data collection was conducted in two phases each of 4 weeks' duration. Any dispensing errors and near misses that occurred during the study periods were recorded by the pharmacy staff in a standard data collection form. A focus group discussion was held with the dispensing staff of participating pharmacies to identify and evaluate the feasibility of the reporting system. Out of a total of 51 357 items dispensed during the two phases of the study, 39 dispensing errors (0.08%) and 247 near misses (0.48%) were detected. The results show that near misses occurred six times more often than dispensing errors, indicating the importance of final checking in pharmacies. The most common types of dispensing errors or near misses appeared to be incorrect strength of medication, followed by incorrect drug, incorrect quantity, incorrect dosage form and incorrect label. Feedback during the focus group discussion indicated that the outcome of the self-reporting scheme was more important than the incidence of errors or near misses. Participating pharmacies also agreed that the self-reporting scheme used was feasible and they would continue using the scheme although some incentives would be helpful. The quantitative results of this study and the qualitative feedback from the participating pharmacies indicate that the self-reporting scheme used is practical and feasible. PMID- 12908850 TI - Hepatic reactions during treatment of multiple sclerosis with interferon-beta-1a: incidence and clinical significance. AB - BACKGROUND: Hepatic dysfunction, manifested as liver enzyme elevations, occurs frequently in patients who are treated with interferon, however, data for patients with multiple sclerosis are limited. OBJECTIVE: To retrospectively assess the safety profile of interferon-beta-1a therapy with respect to liver function during clinical trials and postmarketing surveillance in the treatment of multiple sclerosis. PATIENTS AND METHODS: Adverse effects and laboratory abnormalities were analysed from six randomised, controlled clinical trials (five of which were placebo-controlled) that assessed the use of interferon-beta-1a in patients with multiple sclerosis. Treatment data were collected for 2819 patients for up to 12 months, of whom 1995 received interferon-beta-1a (337 [12%] received Avonex intramuscular therapy, and 1658 [59%] received Rebif subcutaneous therapy), and 824 (29%) received placebo. Data for 2 years were collated for 1178 patients (from two studies). Total weekly interferon doses were 22-132 microg. Postmarketing surveillance data were also analysed. RESULTS: In patients receiving interferon-beta-1a, there were significant elevations of alanine aminotransferase (ALT) levels, of all grades of severity, in up to 59% of patients at 6 months, up to 64% of patients at 12 months and up to 67% of patients at 24 months; ALT elevations were asymptomatic and dose related. More than 50% of elevations in liver enzymes occurred during the first 3 months of treatment, and more than 75% occurred during the first 6 months. Elevated enzyme levels resolved spontaneously or with dosage adjustment. Although the overall incidence of liver enzyme elevation was high during the early months of therapy, after 2 years, the proportion of patients with abnormal liver enzyme levels was 11% of those receiving Rebif 44 microg three times weekly compared with 6% of placebo-treated patients. Only 0.4% of patients discontinued interferon-beta-1a treatment because of hepatic adverse effects. Serious symptomatic interferon related hepatic toxicity occurs, but is uncommon. Concomitant medication use was not associated with increased risk. CONCLUSION: Asymptomatic hepatic dysfunction is common in patients with multiple sclerosis who are treated with interferon beta-1a, and is dose related. Adverse effects are mainly mild and transient, with little impact on adherence to therapy, although rare serious events can occur. Regular liver function monitoring during the first 6 months is recommended. PMID- 12908851 TI - Drug delivery systems for treatment of systemic hypertension. AB - Novel drug delivery systems are available in many areas of medicine. Their application in the treatment of hypertension continues to widen. Oral drug delivery systems permit antihypertensive agents that were previously administered two to four times daily to be administered once daily. Biotechnical use of chemical-dispensing systems has been applied to propranolol (polymer coated beads), clonidine (transdermal therapeutic system), nifedipine (osmotic pump and coat-core), isradipine (osmotic pump), verapamil (sodium alginate and spheroidal oral delivery absorption system), felodipine (coat-core), nisoldipine (coat-core) and diltiazem (polymer coated beads and Geomatrix. The initial goal was to lower blood pressure by a uniform amount throughout the entire day. Now, new drug delivery systems are being developed to target blood pressure in the early morning hours when most cardiovascular events occur. Two chronotherapeutic drug delivery systems are now available for verapamil (chronotherapeutic oral delivery absorption system and delayed coat osmotic pump). Disadvantages of sustained release products include delayed achievement of pharmacodynamic effect, unpredictable bioavailability, enhanced first-pass hepatic metabolism, dose dumping, sustained toxicity, dosage inflexibility and increased cost. Potential advantages include reduced administration frequency, enhanced adherence and convenience, reduced toxicity, stable drug concentrations, uniform drug effect, decreased cost (occasionally) and decreased daily dosage. PMID- 12908852 TI - Pharmacokinetics of L-carnitine. AB - L-Carnitine is a naturally occurring compound that facilitates the transport of fatty acids into mitochondria for beta-oxidation. Exogenous L-carnitine is used clinically for the treatment of carnitine deficiency disorders and a range of other conditions. In humans, the endogenous carnitine pool, which comprises free L-carnitine and a range of short-, medium- and long-chain esters, is maintained by absorption of L-carnitine from dietary sources, biosynthesis within the body and extensive renal tubular reabsorption from glomerular filtrate. In addition, carrier-mediated transport ensures high tissue-to-plasma concentration ratios in tissues that depend critically on fatty acid oxidation. The absorption of L carnitine after oral administration occurs partly via carrier-mediated transport and partly by passive diffusion. After oral doses of 1-6g, the absolute bioavailability is 5-18%. In contrast, the bioavailability of dietary L-carnitine may be as high as 75%. Therefore, pharmacological or supplemental doses of L carnitine are absorbed less efficiently than the relatively smaller amounts present within a normal diet.L-Carnitine and its short-chain esters do not bind to plasma proteins and, although blood cells contain L-carnitine, the rate of distribution between erythrocytes and plasma is extremely slow in whole blood. After intravenous administration, the initial distribution volume of L-carnitine is typically about 0.2-0.3 L/kg, which corresponds to extracellular fluid volume. There are at least three distinct pharmacokinetic compartments for L-carnitine, with the slowest equilibrating pool comprising skeletal and cardiac muscle.L Carnitine is eliminated from the body mainly via urinary excretion. Under baseline conditions, the renal clearance of L-carnitine (1-3 mL/min) is substantially less than glomerular filtration rate (GFR), indicating extensive (98-99%) tubular reabsorption. The threshold concentration for tubular reabsorption (above which the fractional reabsorption begins to decline) is about 40-60 micromol/L, which is similar to the endogenous plasma L-carnitine level. Therefore, the renal clearance of L-carnitine increases after exogenous administration, approaching GFR after high intravenous doses. Patients with primary carnitine deficiency display alterations in the renal handling of L carnitine and/or the transport of the compound into muscle tissue. Similarly, many forms of secondary carnitine deficiency, including some drug-induced disorders, arise from impaired renal tubular reabsorption. Patients with end stage renal disease undergoing dialysis can develop a secondary carnitine deficiency due to the unrestricted loss of L-carnitine through the dialyser, and L-carnitine has been used for treatment of some patients during long-term haemodialysis. Recent studies have started to shed light on the pharmacokinetics of L-carnitine when used in haemodialysis patients. PMID- 12908854 TI - Influence of sex on the pharmacokinetic interaction of fleroxacin and ciprofloxacin with caffeine. AB - BACKGROUND: Previous pharmacokinetic studies have shown that a number of the quinolones inhibit the metabolism of caffeine. OBJECTIVE: To evaluate the effect of sex on the interaction between two quinolones and caffeine. DESIGN: Multiple dose, double-blind, randomised, three-period crossover study. PARTICIPANTS: Twelve male and twelve female healthy volunteers. METHODS: Subjects received by mouth either fleroxacin 400 mg once daily and caffeine 100 mg three times daily, ciprofloxacin 500 mg twice daily and caffeine 100 mg three times daily, or caffeine alone, for 3 days. Subjects received each of the other regimens after 12 day washout periods. Plasma and urine concentrations were determined by validated high-performance liquid chromatography procedures and the data were analysed by noncompartmental linear pharmacokinetic methods. RESULTS: Analysis of the interaction by sex revealed that females showed a significant difference in caffeine pharmacokinetics in the presence of ciprofloxacin (area under the concentration-time curve [AUC], peak plasma concentration [C(max)], time to C(max) [t(max)] and apparent total body clearance [CL/F]) and fleroxacin (AUC and CL/F) when compared with males. Significant differences between sexes were also observed in the pharmacokinetics of ciprofloxacin (AUC, elimination rate constant [beta] and CL/F) and fleroxacin (C(max) and beta) in the presence of caffeine. However, these significant differences disappeared when AUC and C(max) were normalised to 70 kg bodyweight and CL/F was expressed as per kg bodyweight. CONCLUSION: The effect of quinolones on the pharmacokinetics of caffeine, and the reciprocal effect, are different between the sexes, due in part to different bodyweights. PMID- 12908853 TI - Factors affecting the clinical development of cytochrome p450 3A substrates. AB - The objective of this review is to evaluate the risks associated with the discovery and development of cytochrome p450 (CYP) 3A substrates. CYP3A is the most abundant p450 enzyme in human liver and is highly expressed in the intestinal tract. The enzyme contributes substantially to metabolism of approximately 50% of currently marketed drugs that undergo oxidative metabolism. As a result, drug-drug interactions involving inhibitors of CYP3A-mediated metabolism can be of great clinical consequence. It is the position of the authors that, because of the factors responsible for the broad substrate specificity of CYP3A, discovery and development of compounds across a large and broad portfolio that are completely devoid of CYP3A metabolism is not feasible. Thus, it is important that scientifically valid approaches to the discovery and development of compounds metabolised by CYP3A be realised. The clinical relevance of CYP3A metabolism is dependent on a multitude of factors that include the degree of intestinal and hepatic CYP3A-mediated first-pass extraction, the therapeutic index of the compound and the adverse event associated with inhibition of CYP3A metabolism. Thus, a better understanding of the disposition of a CYP3A-metabolised compound relative to the projected or observed therapeutic index (or safety margin) can provide ample evidence to support the continued development of a CYP3A substrate. This document will highlight current practices as well as the benefits and risks associated with those practices. PMID- 12908855 TI - Collagen gel contraction serves to rapidly distinguish epithelial- and mesenchymal-derived cells irrespective of alpha-smooth muscle actin expression. AB - Mesenchymal-like cells in the stroma of breast cancer may arise as a consequence of plasticity within the epithelial compartment, also referred to as epithelial mesenchymal transition, or by recruitment of genuine mesenchymal cells from the peritumoral stroma. Cells of both the epithelial compartment and the stromal compartment express alpha smooth muscle actin (alpha-sm actin) as part of a myoepithelial or a myofibroblastic differentiation program, respectively. Moreover, because both epithelial- and mesenchymal-derived cells are nontumorigenic, other means of discrimination are warranted. Here, we describe the contraction of hydrated collagen gels as a rapid functional assay for the distinction between epithelial- and mesenchymal-derived stromal-like cells irrespective of the status of alpha-sm actin expression. Three epithelial-derived cell lines and three genuine mesenchymal-derived breast cell lines were plated on top of hydrated collagen lattices. Reduction in gel height was measured every hour for 6 h and after 22 h using an x-y-z automated position table. Significantly, the epithelial-derived cells, irrespective of a high alpha-sm actin expression, had a fivefold lower contractility (10.0% reduction in gel height) than their true mesenchymal counterparts (53.1% reduction in gel height). To test whether at all force generation could be induced in the nonmesenchymal cells by alpha-sm actin, transductions were performed to obtain a tetracycline dependent expression. Expression under these conditions did not augment contractility. It is concluded that epithelial-derived mesenchymal-like cells are functionally defective within a connective tissue environment irrespective of an apparent contractile phenotype. PMID- 12908856 TI - Indoor health: background levels of fungi. AB - There is no uniformity in the suggested guidelines for acceptable levels of fungi in indoor ambient air. Thus, health professionals have no way to determine what levels of fungi may pose a threat to human health. The authors reviewed the published literature to identify data reported for noncomplaint structures, that is, structures in which occupants did not have health concerns associated with the quality of the indoor air. For both commercial and residential structures, fungal concentrations detected were often higher than currently suggested guidance values. The average indoor air concentration in 149 noncomplaint commercial buildings was 233 colony forming units (CFU) per cubic meter, whereas outdoor ambient air levels averaged 983 CFU/m(3). Total indoor spore counts ranged from 610 to 1040 spores/m(3) in three commercial buildings. Outdoor total spore counts associated with these buildings ranged from 400 to 80,000 spores/m(3). The average indoor concentration reported for 820 noncomplaint residential structures was 1252 CFU/m(3) with an average outdoor level of 1524 CFU/m(3). Total spore counts detected indoors at 85 residential structures ranged from 68 to 2307 spores/m(3). Outdoor spore levels associated with these structures ranged from 400 to 80,000 spores/m(3). A large proportion of both commercial and residential noncomplaint structures have indoor ambient air fungal concentrations above 500 CFU/m(3), a level often advocated as requiring remediation in structures when occupants complain of nonspecific adverse health symptoms. PMID- 12908857 TI - An HDI polyisocyanate aerosol exposure system for large-scale animal experiments. AB - An exposure system that allows large-scale exposure of animals to 1,6 hexamethylene diisocyanate (HDI)-based polyisocyanates at a stable concentration and aerosol size distribution was developed. The HDI polyisocyanate aerosol is generated by nebulizing a solution of a commercial polyisocyanate product dissolved in acetone. The aerosol is delivered with a constant airflow into a horizontal flow chamber. Complete mixing of aerosol in the chamber is ensured by a circulating fan. This method has been used to generate atmospheres containing HDI polyisocyanates at a concentration of 10.46+/-0.23 mg/m(3) over a 5-hour period. The overall mass median aerodynamic equivalent diameter was found to be 1.42 microm with a geometric standard deviation of 1.26. The HDI monomer concentration was 0.15+/-0.04 mg/m(3). The average chamber acetone concentration was determined to be 2481+/-222 ppm (mean+/-standard deviation). Different HDI polyisocyanate concentrations in the chamber can be achieved by altering the concentration of the commercial polyisocyanate product in acetone and the chamber flow rate. The described exposure system will be useful for performing toxicological studies involving HDI polyisocyanates. PMID- 12908858 TI - Predicting room vapor concentrations due to spills of organic solvents. AB - Relatively small spills of volatile liquids can result in short-term, high concentration exposures. Because of the transient nature of these exposures, air sampling may be precluded. As an alternative, exposure assessment can be done by mathematical modeling. The vapor emission rate from small spills is highest immediately following the spill and decreases as the surface area available for mass transfer decreases and evaporation cools the liquid. This decreasing emission rate is not described by any of the existing evaporation rate models. The authors present an evaporation rate model that describes the changing emissions as exponentially decreasing. The rate of decrease is governed by an evaporation rate parameter alpha, which has the unit of min(-1) and can be estimated based on experimental measurements. The authors measured alpha for a suite of compounds and different sizes of spill. They found that alpha can be estimated for hydrocarbons containing only C, H, and O with the equation: alpha=0.000524 VP + 0.0108 SA/Vol, where VP is the vapor pressure of the liquid and SA/Vol is the surface area to volume ratio. Next, the authors integrated the exponentially decreasing emission rate into a well-mixed room versus a near field/far field dispersion construct to predict vapor concentrations. A preliminary experiment was conducted in a test room to compare measured concentrations with the concentrations predicted by the models. The well-mixed room model performed well based on ANSI indoor air model evaluation criteria. The predicted near field concentrations showed a poor fit to the measured values based on the ANSI criteria, although overall they did capture the observed time profile. PMID- 12908859 TI - Visualization of airflows in push-pull ventilation systems applied to surface treatment tanks. AB - A pilot installation was designed that simulates a surface treatment tank fitted with a push-pull ventilation system. The installation contained elements for measuring and controlling the operational variables (flow rate and tank temperature) and smoke generating equipment for injecting smoke through the holes of the push unit and from the tank surface. Visual observation and video recording of the flows involved meant it was possible to follow the qualitative behavior of the push flow rate along the tank surface and to identify any emissions not captured by the exhaust system. It was possible to differentiate the initial semifree push curtain, its impact with the tank surface, the wall jet that moved toward the exhaust, and its entrance into the exhaust. The methodology proposed is complemented by a quantitative technique for measuring the efficiency, using sulfur hexafluoride as tracer, which permits the causes and location of losses in the ventilation system to be determined. PMID- 12908860 TI - Airway responsiveness of rabbits after exposure to 2-octyl dodecanol. AB - Cooling lubricants are used in the metal industry during drilling or turning. Vapors and aerosols of these lubricants are suspected to induce airway hyperresponsiveness (AHR) in exposed workers. In a previous study the authors demonstrated that water-soluble lubricants induce AHR after acute exposure of rabbits to concentrations near the German MAK value (10 mg/m(3)). In the present investigation the influence of a fatty alcohol as special non-water-soluble cooling lubricant was examined to determine its influence on airway responsiveness (AR). The effects of an aerosolized non-water-soluble lubricant (40, 90, and 220 mg/m(3)) on AR to acetylcholine in a rabbit model were studied. Lubricant atmosphere analysis was performed with infrared spectroscopy. Before exposure, after 2 and 4 hours of application, AR to aerosols from 0.2 and 2% acetylcholine was tested. Basal airway and cardiovascular parameters as well as blood gases did not change during exposure. Lubricant aerosol concentration of 40 and 220 mg/m(3) for 4 hours did not significantly alter AR. Inhalation of 90 mg/m(3) lubricant increased contractile response to ACH significantly. In contrast to formerly investigated water-soluble cooling lubricants, the examined non-water-soluble lubricant did not increase AR in concentrations near the MAK; however, in higher concentrations a significant (p<.05) increase was obtained. PMID- 12908861 TI - Job categories and their effect on exposure to fungal alpha-amylase and inhalable dust in the U.K. baking industry. AB - Enzymes in flour improver, in particular fungal alpha-amylase, are known to be a significant cause of respiratory allergy in the baking industry. This study measured total inhalable dust and fungal alpha-amylase exposures in U.K. bakeries, mills, and a flour improver production and packing facility and determined whether assignment of job description could identify individuals with the highest exposures to fungal alpha-amylase and inhalable dust. A total of 117 personal samples were taken for workers in 19 bakeries, 2 mills, and a flour improver production and packing facility and were analyzed using a monoclonal based immunoassay. Occupational hygiene surveys were undertaken for each site to assign job description and identify individuals who worked directly with flour improvers. Analysis of exposure data identified that mixers and weighers from large bakeries had the highest exposures to both inhalable dust and fungal alpha amylase among the different categories of bakery workers (p<.01). Currently, the maximum exposure limit for flour dust in the United Kingdom is 10 mg/m(3) (8-hour time-weighted average reference period). In this study 25% of the total dust results for bakers exceeded 10 mg/m(3), and interestingly, 63% of the individuals with exposure levels exceeding 10 mg/m(3) were weighers and mixers. Individuals who worked directly with flour improvers were exposed to higher levels of both inhalable dust and fungal alpha-amylase (p<.01) than those who were not directly handling these products. Before sensitive immunoassays were utilized for the detection of specific inhalable allergens, gravimetric analysis was often used as a surrogate. There was a weak relationship between inhalable dust and fungal alpha-amylase exposures; however, inhalable dust levels could not be used to predict amylase exposures, which highlights the importance of measuring both inhalable dust and fungal alpha-amylase exposures. PMID- 12908862 TI - Exposures to diesel exhaust in the International Brotherhood of Teamsters, 1950 1990. AB - A prior case-control study found a positive, monotonic exposure-response relationship between exposure to diesel exhaust and lung cancer among decedents of the Central States Conference of the International Brotherhood of Teamsters. In response to critiques of the Teamsters' exposure estimates by the Health Effects Institute's Diesel Epidemiology Panel, historical exposures and associated uncertainties are investigated here. Historic diesel exhaust exposures are predicted as a function of heavy-duty diesel truck emissions, increasing use of diesel engines, and occupational elemental carbon (EC) measurements taken during the late 1980s and early 1990s. EC from diesel and nondiesel sources is distinguished in light of recent studies indicating a substantial contribution of gasoline vehicles to ambient EC. Monte Carlo sampling is used to characterize exposure distributions. The methodology used in this article-a probabilistic model for historical exposure assessment-is novel. PMID- 12908863 TI - Evaluation of a quantitative fit testing method for N95 filtering facepiece respirators. AB - A method for performing quantitative fit tests (QNFT) with N95 filtering facepiece respirators was developed by earlier investigators. The method employs a simple clamping device to allow the penetration of submicron aerosols through N95 filter media to be measured. The measured value is subtracted from total penetration, with the assumption that the remaining penetration represents faceseal leakage. The developers have used the clamp to assess respirator performance. This study evaluated the clamp's ability to measure filter penetration and determine fit factors. In Phase 1, subjects were quantitatively fit-tested with elastomeric half-facepiece respirators using both generated and ambient aerosols. QNFT were done with each aerosol with both P100 and N95 filters without disturbing the facepiece. In Phase 2 of the study elastomeric half facepieces were sealed to subjects' faces to eliminate faceseal leakage. Ambient aerosol QNFT were performed with P100 and N95 filters without disturbing the facepiece. In both phases the clamp was used to measure N95 filter penetration, which was then subtracted from total penetration for the N95 QNFT. It was hypothesized that N95 fit factors corrected for filter penetration would equal the P100 fit factors. Mean corrected N95 fit factors were significantly different from the P100 fit factors in each phase of the study. In addition, there was essentially no correlation between corrected N95 fit factors and P100 fit factors. It was concluded that the clamp method should not be used to fit-test N95 filtering facepieces or otherwise assess respirator performance. PMID- 12908864 TI - Exposure to particulates, microorganisms, beta(1-3)-glucans, and endotoxins during soybean harvesting. AB - Sclerotinia sclerotiorum is an emerging fungal pathogen affecting soybeans in the United States. In response to its emergence, exposures to particulates, bioaerosols, endotoxins, S. sclerotiorum, and beta(1-3)-glucans were characterized during soybean harvesting. Air sampling was performed on soybean harvesters (combines) and on the farmers in closed cabs as personal samples during harvesting at 17 farms in 1997 and repeated at 15 in 1998. S. sclerotiorum infestation was evident in the fields at 8 of the sites (44%). The geometric mean concentrations (and geometric standard deviations) measured on the combines in 1998 were as follows: total dust, 11.9 (2.8) mg/m(3); inhalable dust 11.7 (6.4) mg/m(3); and beta(1-3)-glucans, 5027 (7) ng/m(3). Values for the personal samples in 1998 were as follows: total dust, 1.2 (6.7) mg/m(3); inhalable dust, 1.1 (5.3); and beta(1-3)-glucans, 674 (9) ng/m(3). These concentrations were two- to threefold higher than in the previous year. Ambient endotoxin concentrations were 4438 EU/m(3) in Year I and 459 EU/m(3) in Year II. Particle size distribution measurements on the combines yielded mass median aerodynamic diameters of 6.6 microm on the combine and 4.0 microm inside the combine cab. Closed combine cabs provided an average workplace protection factor of 11.7 for total dust. Nevertheless, personal exposures to organisms inside combine cabins ranged from 3.6 x 10(4) to 4.0 x 10(8) organisms/m(3). These data indicate the potential exists for high exposures to organic dust and bioaerosols during soybean harvesting. PMID- 12908865 TI - Control of workers' exposure to airborne endotoxins and formaldehyde during the use of metalworking fluids. AB - The study evaluated the effects of triazine use and machine enclosure on workers' exposure to bacteria, endotoxins, and formaldehyde in the use of metalworking fluids (MWF). Concentrations of triazine and bacterial contaminants were monitored in bulk samples of MWF from two machines in one workplace. One of the machines was used normally; triazine was added to the other when needed. Air sampling was used to estimate workers' exposure to endotoxins at 18 workplaces near enclosed and open machines. Concentrations of triazine in MWF and formaldehyde in the air were measured. Some recirculating local exhaust ventilation systems were also tested. The endotoxin and bacteria concentrations in the biocide test of MWF rapidly increased when the biocide levels decreased below 500 ppm. Airborne concentrations of endotoxins were substantially lower near enclosed machines than near open ones. Concentrations of airborne formaldehyde were below the Finnish occupational exposure limit at the existing levels of triazine in MWF. Concentrations of triazine in MWF correlated well with those of formaldehyde in the air near the machines (correlation coefficient r=.69). The results showed that the triazine levels in MWF should continuously be kept high enough (>500 ppm) to prevent workers' exposure to endotoxins and bacteria. Overdosing with triazine, however, should be avoided, so that the levels of airborne formaldehyde remain low. Triazine levels in MWF can be followed by the method described in this article. Workers' exposure to contaminants of MWF can be reduced substantially by enclosing the machines and equipping them with local exhausts. PMID- 12908866 TI - Alternative field methods for measuring hearing protector performance. AB - In comparison with the mandatory noise reduction rating (NRR) testing of every hearing protector sold in the United States, real-world tests of hearing protector attenuation are scarce. This study evaluated data from three potential field-test methods as compared with the subject-fit data from Method B of ANSI S12.6-1997 for the E.A.R(R) Express trade mark Pod Plug trade mark. The new field test methods were the FitCheck headphone (FCH) method, FitCheck in sound field (FCSF) method, and bone-conduction loudness balance (BCLB) method, all of which can be administered in small single-person audiometric booths such as are commonly found in industry. Twenty normal-hearing and audiometrically competent subjects naive to hearing protector use were tested with the laboratory and the three field-test methods in a repeated-measures design. Repeated-measures models with structured covariance matrices were used to analyze the data. Significant effects were found for method, frequency, and first-order frequency-by-gender and frequency-by-method interactions. These effects and interactions were expected given the different psychophysical tasks. The FCSF and BCLB methods provided attenuations that were not significantly different from those found with Method B. Although the attenuations measured for the FCH method were statistically different (greater) than the attenuations from the other methods, the differences were within the magnitude of acceptable test-retest audiometric variability. The results suggest that the FCH and FCSF methods were both feasible and reliable methods for field testing. The FCH method is limited to testing earplugs, and the FCSF requires additional equipment to outfit the test booth, but could be used for testing all types of protectors. PMID- 12908867 TI - Upper body cooling during exercise-heat stress wearing the improved toxicological agent protective system for HAZMAT operations. AB - This study compared endurance in a U.S. Army developmental Occupational Safety and Health Administration Level B personal protective equipment (PPE) system against the toxicological agent protective (TAP) suit, the Army's former standard PPE for Level A and Level B toxic environments. The developmental system consisted of two variations: the improved toxicological agent protective (ITAP) suit with self-contained breathing apparatus (ITAP-SCBA), weight 32 kg, and the ITAP with blower (ITAP-B), weight 21 kg. Both ITAP suits included the personal ice cooling system (PICS). TAP (weight 9.5 kg) had no cooling. It was hypothesized that PICS would effectively cool both ITAP configurations, and endurance in TAP would be limited by heat strain. Eight subjects (six men, two women) attempted three 2-hour treadmill walks (0.89 m/sec, 0% grade, rest/exercise cycles of 10/20 min) at 38 degrees C, 30% relative humidity. Metabolic rate for TAP (222+/-35 W) was significantly less than either ITAP-SCBA (278+/-27 W) or ITAP-B (262+/-24 W) (p<0.05). Endurance time was longer in ITAP SCBA (85+/-20 min) and ITAP-B (87+/-25 min) than in TAP (46+/-10 min) (p<0.05). Heat storage was greater in TAP (77+/-15 W.m(-2)) than in ITAP-SCBA (51+/-16 W.m( 2)) (p<0.05), which was not different from ITAP-B (59+/-14 W.m(-2)). Sweating rate was greater in TAP (23.5+/-11.7 g/min(1)) than in either ITAP-SCBA (11.1+/ 2.9 g/min) or ITAP-B (12.8+/-3.5 g/min) (p<0.05). Endurance in ITAP was nearly twice as long as in PPE with no cooling, even though the PICS, SCBA tanks, and new uniform itself all served to increase metabolic cost over that in TAP. PICS could also be used with civilian Levels A and B PPE increasing work time and worker safety. PMID- 12908868 TI - Workplace exposure to submicron particle mass and number concentrations from manual arc welding of carbon steel. AB - Particle emissions from manual shielded metal arc welding of carbon steel were sampled in a typical industrial maintenance and metal fabrication workplace environment. Particle number measurements over the size range from 14 nm to 10 microm using a scanning mobility particle sizer and an optical particle counter showed that welding produced an approximately lognormal particle mode with a 120 nm count median and a geometric standard deviation of 2.07. This study produced welding particle number concentrations on the order of 2 x 10(5)/cm(3) in the building air 8.5 m away from the welding. Workplace exposure samples were below the current 8-hour American Conference of Governmental Industrial Hygienists mass concentration threshold limit value of 5 mg/m(3). Submicron particles comprised 80% of the total aerosol mass collected by a cascade impactor during welding. The concentration of larger particles was indistinguishable from indoor background. Microscopy showed that the welding emissions are dominated by clusters formed from <0.1 microm primary spheres. These data on the particles resulting from aerosol transformation by natural dilution inside an industrial building can be compared with laboratory-scale studies of welding particulate. The particle number characteristics observed in this study are significant because toxicological hypotheses suggest that number or surface area may be a better metric than mass when evaluating the health effects of fine particles. PMID- 12908869 TI - Particle size distribution and respiratory deposition estimates of beryllium aerosols in an extraction and processing plant. AB - The mass size distribution of beryllium aerosols generated in the various operational areas of a typical extraction and processing plant was studied using an eight-stage impactor sampler. The total concentration of beryllium in the plant was found to be well below the threshold limit value. The mean value of mass median aerodynamic diameter of beryllium particles observed for various operations ranged from 5.0-9.5 microm. The alveolar deposition for various operational areas was estimated to be 3-5% for nasal breathing and 9-13% for oral breathing based on the International Commission on Radiological Protection (ICRP) human respiratory tract model. Deposition during oral breathing was higher than during nasal breathing by approximately a factor of two to three. This study on exposure characterization was useful for reducing the respirable fraction of beryllium aerosol by optimizing the capture velocity and improving the quality of other control measures. PMID- 12908870 TI - Field evaluation and comparison of five methods of sampling lead dust on carpets. AB - Five methods of sampling lead-contaminated dust on carpets were evaluated and compared in 33 New Jersey homes of children with elevated blood lead levels. The five sampling methods were (1) wipe, (2) adhesive label, (3) C18 sheet, (4) vacuum, and (5) hand rinse. Samples were collected side by side on the same carpets within the homes. Among the five methods the wipe and vacuum methods showed high percentages of detectable samples, good reproducibility, and significant correlations with other methods. C18 sheets and adhesive labels collected the least quantity of lead dust, with high percentages of undetectable samples. Because of the limited ability of sampling lead on carpets and the relatively high cost for laboratory analysis, C18 sheets or adhesive labels are not considered feasible sampling techniques. The hand rinse method also was not feasible for carpet sampling, because it was difficult to conduct in the field and laboratory, and it was subject to inconsistency and cross contamination. Wipes, which collected lead dust from carpet surfaces, were believed to be the most appropriate method for measuring lead from carpets accessible to children. However, because of the low pickup from carpets, wipes may not be an appropriate measuring tool to assess the levels of total lead contamination in carpets. The authors recommend using surface wipe sampling to measure accessible lead from carpets for exposure assessment, and vacuum sampling to obtain the information on total lead accumulation. PMID- 12908871 TI - Surface germicidal effects of ozone for microorganisms. AB - In this study the influences of microorganism species, relative humidity, and ozone dosage on ozone surface disinfection were evaluated. Bacterial and fungal cultures were spread on agar plates and exposed to ozone. The selected microorganisms included Escherichia coli, Bacillus subtilis, Candida famata, and Penicillium citrinum. Results showed that microorganism survival fraction and ozone dosage (ozone concentration times exposure time) have an exponential relationship. Results also indicated that E. coli was the most sensitive organism to ozone exposure. E. coli required only very low ozone doses of 2-2.5 and 3.5-4 mg to obtain 50 and 80% inactivation, respectively. In addition, P. citrinum was more resistant than E. coli and required ozone doses of 40-60 and 60-120 mg to obtain 50 and 80% inactivation. In addition, spores of B. subtilis were observed to be the most resistant organism, requiring ozone doses of 40-75 and 145-150 mg to obtain 50 and 80% inactivation. Yeast was less resistant than P. citrinum and B. subtilis, requiring ozone doses of 10 and 15-19 mg to obtain 50 and 80% inactivation. It was clearly indicated that the ozone dose differences for 80% microorganism inactivation could be as high as 40 times between B. subtilis and E. coli. Ozone surface germicidal efficiency increased as relative humidity increased, which could be related to more radicals generated from ozone reaction with more water vapor at higher relative humidity. It was concluded that ozone should be highly effective and provide a reliable safety factor in treating contaminated surface. In addition, workers might need to wear suitable respiratory protection at high ozone level operation. PMID- 12908872 TI - Site-directed mutagenesis of the active site of diacylglycerol kinase alpha: calcium and phosphatidylserine stimulate enzyme activity via distinct mechanisms. AB - Diacylglycerol kinases (DAGKs) catalyse ATP-dependent phosphorylation of sn-1,2 diacylglycerol that arises during stimulated phosphatidylinositol turnover. DAGKa is activated in vitro by Ca2+ and by acidic phospholipids. The regulatory region of DAGKa includes an N-terminal RVH motif and EF hands that mediate Ca2+ dependent activation. DAGKa also contains tandem C1 protein kinase C homology domains. We utilized yeast, Saccharomyces cerevisiae, which lacks an endogenous DAGK, to express DAGKa and to determine the enzymic activities of different mutant forms of pig DAGKa in vitro. Six aspartate residues conserved in all DAGKs were individually examined by site-directed mutagenesis. Five of these aspartate residues reside in conserved blocks that correspond to sequences in the catalytic site of phosphofructokinases. Mutation of D434 (Asp434) or D650 abolished all DAGKa activity, whereas substitution of one among D465, D497, D529 and D697 decreased the activity to 6% or less of that for wild-type DAGKa. Roles of homologous residues in phosphofructokinases suggested that the N-terminal half of the DAGK catalytic domain binds Mg-ATP and the C-terminal half binds diacylglycerol. A DAGKa mutant with its entire regulatory region deleted showed a much decreased activity that was not activated by Ca2+, but still exhibited PS (phosphatidylserine)-dependent activation. Moreover, mutations of aspartate residues at the catalytic domain had differential effects on activation by Ca2+ and PS. These results indicate that Ca2+ and PS stimulate DAGKa via distinct mechanisms. PMID- 12908873 TI - Rapid agonist-evoked coupling of type II Ins(1,4,5)P3 receptor with human transient receptor potential (hTRPC1) channels in human platelets. AB - Depletion of intracellular Ca2+ stores results in the activation of SMCE (store mediated Ca2+ entry) in many cells. The mechanism of activation of SMCE is poorly understood. In human platelets, a secretion-like coupling model may be involved. This proposes that store depletion results in trafficking of portions of the endoplasmic reticulum to the plasma membrane, enabling coupling between proteins in the two membranes. In support of this, we have shown that, in human platelets, agonist-evoked Ca2+ store depletion results in de novo and reversible coupling of the Ins P3RII [type II inositol (1,4,5)trisphosphate receptor] with the putative Ca2+ entry channel hTRPC1 [human canonical transient receptor potential 1 (protein); Rosado, Brownlow and Sage (2002) J. Biol. Chem. 277, 42157-42163]. A crucial test of the hypothesis that this coupling activates SMCE is that it should occur rapidly enough to account for agonist-evoked Ca2+ entry. In the present study, we have used quenched- and stopped-flow approaches to determine the latencies of thrombin-evoked coupling of Ins P3RII with hTRPC1 and of thrombin-evoked bivalent cation entry using Mn2+ quenching of fura 2 fluorescence. Thrombin-evoked Mn2+ entry was detected with a latency of 0.81+/ 0.07 s (S.E.M., n =7) or 1.36+/-0.09 s (S.E.M., n =7) at a concentration of 1.0 or 0.1 unit/ml respectively. Coupling between Ins P3RII and hTRPC1, assessed at 100 ms intervals, was first detected with a latency of 0.9 or 1.4 s after stimulation with thrombin at a concentration of 1.0 or 0.1 unit/ml respectively. These results support the hypothesis that de novo coupling of Ins P3RII with hTRPC1 could activate SMCE in human platelets. PMID- 12908875 TI - Familial Mediterranean fever, inflammation and nephrotic syndrome: fibrillary glomerulopathy and the M680I missense mutation. AB - BACKGROUND: Familial Mediterranean fever (FMF) is an autosomal recessive disease characterized by inflammatory serositis (fever, peritonitis, synovitis and pleuritis). The gene locus responsible for FMF was identified in 1992 and localized to the short arm of chromosome 16. In 1997, a specific FMF gene locus, MEFV, was discovered to encode for a protein, pyrin that mediates inflammation. To date, more than forty missense mutations are known to exist. The diversity of mutations identified has provided insight into the variability of clinical presentation and disease progression. CASE REPORT: We report an individual heterozygous for the M680I gene mutation with a clinical diagnosis of FMF using the Tel-Hashomer criteria. Subsequently, the patient developed nephrotic syndrome with biopsy-confirmed fibrillary glomerulonephritis (FGN). Further diagnostic studies were unremarkable with clinical workup negative for amyloidosis or other secondary causes of nephrotic syndrome. DISCUSSION: Individuals with FMF are at greater risk for developing nephrotic syndrome. The most serious etiology is amyloidosis (AA variant) with renal involvement, ultimately progressing to end stage renal disease. Other known renal diseases in the FMF population include IgA nephropathy, IgM nephropathy, Henoch-Schonlein purpura as well as polyarteritis nodosa. CONCLUSION: To our knowledge, this is the first association between FMF and the M680I mutation later complicated by nephrotic syndrome and fibrillary glomerulonephritis. PMID- 12908874 TI - The Drosophila forkhead transcription factor FOXO mediates the reduction in cell number associated with reduced insulin signaling. AB - BACKGROUND: Forkhead transcription factors belonging to the FOXO subfamily are negatively regulated by protein kinase B (PKB) in response to signaling by insulin and insulin-like growth factor in Caenorhabditis elegans and mammals. In Drosophila, the insulin-signaling pathway regulates the size of cells, organs, and the entire body in response to nutrient availability, by controlling both cell size and cell number. In this study, we present a genetic characterization of dFOXO, the only Drosophila FOXO ortholog. RESULTS: Ectopic expression of dFOXO and human FOXO3a induced organ-size reduction and cell death in a manner dependent on phosphoinositide (PI) 3-kinase and nutrient levels. Surprisingly, flies homozygous for dFOXO null alleles are viable and of normal size. They are, however, more sensitive to oxidative stress. Furthermore, dFOXO function is required for growth inhibition associated with reduced insulin signaling. Loss of dFOXO suppresses the reduction in cell number but not the cell-size reduction elicited by mutations in the insulin-signaling pathway. By microarray analysis and subsequent genetic validation, we have identified d4E-BP, which encodes a translation inhibitor, as a relevant dFOXO target gene. CONCLUSION: Our results show that dFOXO is a crucial mediator of insulin signaling in Drosophila, mediating the reduction in cell number in insulin-signaling mutants. We propose that in response to cellular stresses, such as nutrient deprivation or increased levels of reactive oxygen species, dFOXO is activated and inhibits growth through the action of target genes such as d4E-BP. PMID- 12908876 TI - Safety and immunogenicity of three doses of an eleven-valent diphtheria toxoid and tetanus protein--conjugated pneumococcal vaccine in Filipino infants. AB - BACKGROUND: An 11-valent pneumococcal conjugate vaccine could provide significantly larger reduction in pneumococcal disease burden than the currently available 7-valent vaccine formulation in many countries. METHODS: In total, 50 infants were enrolled to this open, uncontrolled study, which evaluated the safety and immunogenicity of an aluminium adjuvanted 11-valent mixed-carrier diphtheria toxoid or tetanus protein-conjugated vaccine (11-PncTD) when administered in three doses at 6, 10 and 14 weeks of age simultaneously with DTwP//PRP-T and OPV vaccines in Filipino infants. RESULTS: The rates of local reactions between the two injection sites, those associated with the 11-PncTD vaccine and those with the DTwP//PRP-T were almost of equal frequency for all three vaccine doses except for induration, which was significantly more common in the DTP//PRP-T injection site. Fever was present in 39%, 22% and 21% of infants following each of the three doses. Antibody responses were determined by an enzyme immunoassay method before the first vaccination and after the three doses. The vaccine elicited a significant anti-pneumococcal polysaccharide antibody response against all serotypes included in the vaccine, except for type 14, for which the pre-vaccination geometric mean antibody concentration (GMC) was high (1.61 microg/ml). The GMCs one month after the vaccination series ranged from 1.1 micrograms/ml for type 6B to 23.4 microg/ml for type 4. CONCLUSION: The 11-PncTD vaccine is safe, well-tolerated and immunogenic. The effectiveness of the non adjuvanted formulation of the vaccine in preventing pneumonia is currently being evaluated in the Philippines. PMID- 12908877 TI - Anatomical location of macrophage migration inhibitory factor in urogenital tissues, peripheral ganglia and lumbosacral spinal cord of the rat. AB - BACKGROUND: Previous work suggested that macrophage migration inhibitory factor (MIF) may be involved in bladder inflammation. Therefore, the location of MIF was determined immunohistochemically in the bladder, prostate, major pelvic ganglia, sympathetic chain, the L6-S1 dorsal root ganglia (DRG) and the lumbosacral spinal cord of the rat. RESULTS: In the pelvic organs, MIF immunostaining was prominent in the epithelia. MIF was widely present in neurons in the MPG and the sympathetic chain. Some of those neurons also co-localized tyrosine hydroxylase (TH). In the DRGs, some of the neurons that stained for MIF also stained for Substance P. In the lumbosacral spinal cord, MIF immunostaining was observed in the white mater, the dorsal horn, the intermediolateral region and in the area around the central canal. Many cells were intensely stained for MIF and glial fibrillary acidic protein (GFAP) suggesting they were glial cells. However, some cells in the lumbosacral dorsal horn were MIF positive, GFAP negative cells suggestive of neurons. CONCLUSIONS: Therefore, MIF, a pro-inflammatory cytokine, is localized to pelvic organs and also in neurons of the peripheral and central nervous tissues that innervate those organs. Changes in MIF's expression at the end organ and at peripheral and central nervous system sites suggest that MIF is involved in pelvic viscera inflammation and may act at several levels to promote inflammatory changes. PMID- 12908878 TI - Molecular cloning, genomic characterization and over-expression of a novel gene, XRRA1, identified from human colorectal cancer cell HCT116Clone2_XRR and macaque testis. AB - BACKGROUND: As part of our investigation into the genetic basis of tumor cell radioresponse, we have isolated several clones with a wide range of responses to X-radiation (XR) from an unirradiated human colorectal tumor cell line, HCT116. Using human cDNA microarrays, we recently identified a novel gene that was down regulated by two-fold in an XR-resistant cell clone, HCT116Clone2_XRR. We have named this gene as X-ray radiation resistance associated 1 (XRRA1) (GenBank BK000541). Here, we present the first report on the molecular cloning, genomic characterization and over-expression of the XRRA1 gene. RESULTS: We found that XRRA1 was expressed predominantly in testis of both human and macaque. cDNA microarray analysis showed three-fold higher expression of XRRA1 in macaque testis relative to other tissues. We further cloned the macaque XRRA1 cDNA (GenBank AB072776) and a human XRRA1 splice variant from HCT116Clone2_XRR (GenBank AY163836). In silico analysis revealed the full-length human XRRA1, mouse, rat and bovine Xrra1 cDNAs. The XRRA1 gene comprises 11 exons and spans 64 kb on chromosome 11q13.3. Human and macaque cDNAs share 96% homology. Human XRRA1 cDNA is 1987 nt long and encodes a protein of 559 aa. XRRA1 protein is highly conserved in human, macaque, mouse, rat, pig, and bovine. GFP-XRRA1 fusion protein was detected in both the nucleus and cytoplasm of HCT116 clones and COS-7 cells. Interestingly, we found evidence that COS-7 cells which over-expressed XRRA1 lacked Ku86 (Ku80, XRCC5), a non-homologous end joining (NHEJ) DNA repair molecule, in the nucleus. RT-PCR analysis showed differential expression of XRRA1 after XR in HCT116 clones manifesting significantly different XR responses. Further, we found that XRRA1 was expressed in most tumor cell types. Surprisingly, mouse Xrra1 was detected in mouse embryonic stem cells R1. CONCLUSIONS: Both XRRA1 cDNA and protein are highly conserved among mammals, suggesting that XRRA1 may have similar functions. Our results also suggest that the genetic modulation of XRRA1 may affect the XR responses of HCT116 clones and that XRRA1 may have a role in the response of human tumor and normal cells to XR. XRRA1 might be correlated with cancer development and might also be an early expressed gene. PMID- 12908879 TI - Study of human SP-A, SP-B and SP-D loci: allele frequencies, linkage disequilibrium and heterozygosity in different races and ethnic groups. AB - BACKGROUND: SP-A, SP-B, and SP-D are pulmonary surfactant proteins. Several linkage and association studies have been done using these genes as markers to locate pulmonary disease susceptibility genes, but few have studied the markers systematically in different ethnic groups. Here we studied eight markers in SP-A, SP-B, and SP-D genes in seven ethnic groups from three races (Caucasian, Black and Hispanic). We measured the similarity of the marker distribution among the ethnic groups in order to see whether people in different ethnic groups or races could be mixed together for linkage and association studies. To evaluate the usefulness of these markers, we estimated the informativeness of each marker loci in the seven ethnic groups by assessing their heterozygosity and PIC values. We also conducted linkage disequilibrium (LD) analysis to identify associated marker loci and to estimate the haplotype frequencies in each of the seven ethnic groups in an attempt to find valuable haplotypes so that the level of polymorphism of the "markers" could be increased. RESULTS: Our findings indicate that allele and genotype frequencies may be different between different ethnic groups, especially between ethnic groups from different races. The markers are in general polymorphic in a variety of study groups, especially for the two SP-A1 and SP-A2 markers. Two-locus LD analysis reveals that three pairs of loci are strongly associated together: B-18(A/C) with B1013(A/C), DA11(C/T) with DA160(A/G), SP-A1 with SP-A2. Three-locus LD analysis suggests that B-18(A/C), B1013(A/C) and B1580(C/T) are strongly associated with each other. CONCLUSIONS: Allele and genotype frequency differences imply that different ethnic groups should be mixed with extreme caution before performing linkage and association studies. The associated markers could be used together to increase the level of polymorphism and the informativeness of the "markers". PMID- 12908880 TI - Diapause in the pea aphid (Acyrthosiphon pisum) is a slowing but not a cessation of development. AB - BACKGROUND: Many insects undergo a period of arrested development, called diapause, to avoid seasonally recurring adverse conditions. Whilst the phenology and endocrinology of insect diapause have been well studied, there has been comparatively little research into the developmental details of diapause. We investigated developmental aspects of diapause in sexually-produced embryos of the pea aphid, Acyrthosiphon pisum. RESULTS: We found that early stages of embryogenesis progressed at a temperature-independent rate, characteristic of diapause, whereas later stages of embryogenesis progressed at a temperature dependent rate. However, embryos maintained at very high temperatures during the temperature-independent stage showed severe developmental abnormalities. Under no temperature regime did embryos display a distinct resting stage. Rather, morphological development progressed slowly but continuously throughout embryogenesis. CONCLUSION: Diapause in the pea aphid, and perhaps in many other insects, is a temperature-independent slowing but not a cessation of morphological development. This suggests that the mechanisms limiting developmental rate during diapause may be the same as those controlling developmental rate at other stages of growth. PMID- 12908884 TI - Dietary guidelines for sugar: the need for evidence. PMID- 12908885 TI - An adaptive metabolic demand model for protein and amino acid requirements. AB - The shortcomings of the metabolic implications of the current protein requirements model are reviewed, and an alternative model, validated with [1 (13)C]leucine balance results in human adults, is presented and evaluated in the context of defining protein requirements. The model identifies metabolic demands for amino acids as comprising a small fixed component and a variable adaptive component that is relatively insensitive to acute food or protein intake, but which changes slowly with a sustained change in intake, enabling N equilibrium to be achieved. The model accounts for the apparent low efficiency of utilisation of animal proteins in N balance studies and enables more realistic efficiency values to be measured within an experimental framework that takes account of the adaptive metabolic demand. However, the complex relationship between the adaptive metabolic demand and habitual level and quality of protein intake prevents prediction of protein quality by amino acid scoring, which can markedly underestimate actual values. In contrast to the current model, for fully adapted individuals risk of deficiency (i.e. negative N balance after complete adaptation) will only start to increase when intakes fall below the range of the true minimum requirements, i.e. a value that is currently unknown, but likely to be between 0.40 and 0.50 g/kg per d at the lower end of the reported distribution of requirements. At intakes greater than this with additional metabolic demands varying directly with intake, deficiency is only likely as a short-term response to a change to a lower intake within the adaptive range. Thus, for adults satisfying energy needs on most mixed human diets, intakes will be within the adaptive range, and N equilibrium ceases to be a useful indicator of nutritional adequacy of protein. In the context of prescriptive dietary guidelines it may be expedient to retain current values until the benefits (and any risks) of protein intakes within the adaptive range can be quantified. However, from a diagnostic perspective, indicators other than N balance need to be identified, since maintenance of N balance can no longer be used as a surrogate of adequate protein related health. PMID- 12908886 TI - Effects of dietary macronutrient content on energy metabolism and uncoupling protein mRNA expression in broiler chickens. AB - The objective of the present study was to investigate the effects of dietary macronutrient ratio on energy metabolism and on skeletal muscle mRNA expression of avian uncoupling protein (UCP), thought to be implicated in thermogenesis in birds. Broiler chickens from 2 to 6 weeks of age received one of three isoenergetic diets containing different macronutrient ratios (low-lipid (LL) 30 v. 77 g lipid/kg; low-protein (LP) 125 v. 197 g crude protein (Nx6.25)/kg; low carbohydrate (LC) 440 v. 520 g carbohydrate/kg). LP chickens were characterised by significantly lower body weights and food intakes compared with LL and LC chickens (-47 and -38 % respectively) but similar heat production/kg metabolic body weight, as measured by indirect calorimetry, in the three groups. However, heat production/g food ingested was higher in animals receiving the LP diet (+41 %, P<0.05). These chickens also deposited 57 % less energy as protein (P<0.05) and 33 % more as fat. No significant differences in energy and N balances were detected between LL and LC chickens. The diets with the higher fat contents (i.e. the LP and LC diets) induced slightly but significantly higher relative expressions of avian UCP mRNA in gastrocnemius muscle, measured by reverse transcription-polymerase chain reaction, than the LL diet (88 and 90 v. 78 % glyceraldehyde-3-phosphate dehydrogenase respectively, P<0.05). Our present results are consistent with the recent view that UCP homologues could be involved in the regulation of lipid utilisation as fuel substrate and provide evidence that the macronutrient content of the diet regulates energy metabolism and especially protein and fat deposition. PMID- 12908887 TI - Whole-body fluxes and partitioning of amino acids to the mammary gland of cows fed fresh pasture at two levels of intake during early lactation. AB - The utilisation of essential amino acids (EAA) by the mammary gland of lactating dairy cows fed fresh forages was studied to provide basic information useful in designing strategies to increase the production of milk protein from pasture-fed dairy cows. The relationship between the flux of EAA in the whole body and their uptake by the mammary gland was determined in four cows in early lactation (length of time in milk 44 (SD 14.5) d) producing 21 (SD 4.0) kg milk/d. The cows were maintained in metabolism stalls and fed fresh perennial ryegrass (Lolium perenne) and white clover (Trifolium repens) pasture ad libitum or restricted to 75 % ad libitum intake. The whole-body fluxes of amino acids (AA) were measured using an arterio-venous infusion of universally (13)C-labelled AA. Whole-body fluxes of fourteen AA were estimated. Isotope dilution indicated that mammary utilisation accounted for one-third of the whole-body flux of EAA, with individual AA ranging between 17 and 35 %. Isoleucine, leucine, valine and lysine were the EAA with the greatest partitioning towards the mammary gland (up to 36 % of the whole-body flux), which could reflect a potentially limiting effect on milk protein synthesis. In the case of AA with low partitioning to the mammary gland (for example, histidine), it is suggested that non-mammary tissues may have priority over the mammary gland and therefore the supply of this AA may also limit milk protein synthesis. PMID- 12908888 TI - Variable maternal nutrition and growth hormone treatment in the second quarter of pregnancy in pigs alter semitendinosus muscle in adolescent progeny. AB - Maternal nutrition and growth hormone (GH) treatment during early- to mid pregnancy can each alter the subsequent growth and differentiation of muscle in progeny. We have investigated the effects of varying maternal nutrition and maternal treatment with porcine (p) GH during the second quarter of pregnancy in gilts on semitendinosus muscle cross-sectional area and fibre composition of progeny, and relationships between maternal and progeny measures and progeny muscularity. Fifty-three Large White x Landrace gilts, pregnant to Large White x Duroc boars, were fed either 2.2 kg (about 35 % ad libitum intake) or 3.0 kg commercial ration (13.5 MJ digestible energy, 150 g crude protein (N x 6.25)/kg DM)/d and injected with 0, 4 or 8 mg pGH/d from day 25 to 50 of pregnancy, then all were fed 2.2 kg/d for the remainder of pregnancy. The higher maternal feed allowance from day 25 to 50 of pregnancy increased the densities of total and secondary fibres and the secondary:primary fibre ratio in semitendinosus muscles of their female progeny at 61 d of age postnatally. The densities of secondary and total muscle fibres in semitendinosus muscles of progeny were predicted by maternal weight before treatment and maternal plasma insulin-like growth factor II during treatment. Maternal pGH treatment from day 25 to day 50 of pregnancy did not alter fibre densities, but increased the cross-sectional area of the semitendinosus muscle; this may be partially explained by increased maternal plasma glucose. Thus, maternal nutrition and pGH treatment during the second quarter of pregnancy in pigs independently alter muscle characteristics in progeny. PMID- 12908889 TI - Influence of grapefruit-, orange- and apple-juice consumption on urinary variables and risk of crystallization. AB - Alkalizing beverages are highly effective in preventing the recurrence of calcium oxalate (Ox), uric acid and cystine lithiasis. The aim of the present study was to evaluate the influence of grapefruit-juice and apple-juice consumption on the excretion of urinary variables and the risk of crystallization in comparison with orange juice. All investigations were carried out on nine healthy female subjects without any history of stone formation and aged 26-35 years. Each juice was tested in a 5 d study. During the study, the subjects received a standardized diet. Fluid intake of 2.75 litres was composed of 2.25 litres neutral mineral water, 0.4 litre coffee and 0.1 litre milk. On the fourth and fifth day 0.5 litre mineral water was partly substituted by 0.5 or 1.0 litre juice under investigation respectively. The influence on urinary variables was evaluated in 24 h urine samples. In addition, the BONN risk index of CaOx, relative supersaturation (RS)CaOx crystallization was determined. Due to an increased pH value and an increased citric acid excretion after consumption of each juice, the RSCaOx decreased statistically significantly (P<0.05) for grapefruit juice, but not significantly for orange and apple juice. The BONN risk index yielded a distinct decrease in the crystallization risk. We showed that both grapefruit juice and apple juice reduce the risk of CaOx stone formation at a magnitude comparable with the effects obtained from orange juice. PMID- 12908890 TI - Carbohydrates digestibility and faecal nitrogen excretion in rats fed raw or germinated faba bean (Vicia faba)- and chickpea (Cicer arietinum)-based diets. AB - Raw or germinated faba bean (Vicia faba minor var. Alameda) and chickpea (Cicer arietinum kabuli var. Athenas) seed meals were incorporated in essential amino acid-supplemented and energy-equalized diets for growing (65 (SD 1) g) male Wistar rats as the only sources of dietary protein. A lactalbumin-based diet was used as the control. Faecal dry weight and N excretion of animals fed legume containing diets were greater (P<0.01) and DM digestibility lower (P<0.01) than controls. Apparent faecal digestibilities of amino acids were found to be not different or lower (P<0.01) than controls in rats fed diets containing legume seeds, either germinated or not. Total diaminopimelic acid (DAPA) and purine bases excretion were significantly (P<0.01) higher than controls in rats fed both legume seed meals. Faecal bacterial N calculated according to DAPA or purine bases values was similar and significantly higher (P<0.01) than controls in rats fed legume seed meals. Bacterial N accounted for 50 to 80 % of total faecal N in rats fed legume diets. Apparent faecal N digestibility values (53-65 %) of rats fed legume-based diets were lower (P<0.01) than controls, but became substantially higher (85-92 %) when calculated taking into account bacterial N values. Faecal NSP digestibility values for legume diets were 40-57 g/100 g ingested. Germination decreased starch faecal excretion (P<0.05) and increased (P<0.05) faecal starch and NSP digestibilities of faba bean seeds. PMID- 12908891 TI - Effect of altered dietary n-3 fatty acid intake upon plasma lipid fatty acid composition, conversion of [13C]alpha-linolenic acid to longer-chain fatty acids and partitioning towards beta-oxidation in older men. AB - The effect of increased dietary intakes of alpha-linolenic acid (ALNA) or eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) for 2 months upon plasma lipid composition and capacity for conversion of ALNA to longer-chain metabolites was investigated in healthy men (52 (SD 12) years). After a 4-week baseline period when the subjects substituted a control spread, a test meal containing [U-(13)C]ALNA (700 mg) was consumed to measure conversion to EPA, docosapentaenoic acid (DPA) and DHA over 48 h. Subjects were then randomised to one of three groups for 8 weeks before repeating the tracer study: (1) continued on same intake (control, n 5); (2) increased ALNA intake (10 g/d, n 4); (3) increased EPA+DHA intake (1.5 g/d, n 5). At baseline, apparent fractional conversion of labelled ALNA was: EPA 2.80, DPA 1.20 and DHA 0.04 %. After 8 weeks on the control diet, plasma lipid composition and [(13)C]ALNA conversion remained unchanged compared with baseline. The high-ALNA diet resulted in raised plasma triacylglycerol-EPA and -DPA concentrations and phosphatidylcholine-EPA concentration, whilst [(13)C]ALNA conversion was similar to baseline. The high (EPA+DHA) diet raised plasma phosphatidylcholine-EPA and -DHA concentrations, decreased [(13)C]ALNA conversion to EPA (2-fold) and DPA (4-fold), whilst [(13)C]ALNA conversion to DHA was unchanged. The dietary interventions did not alter partitioning of ALNA towards beta-oxidation. The present results indicate ALNA conversion was down-regulated by increased product (EPA+DHA) availability, but was not up-regulated by increased substrate (ALNA) consumption. This suggests regulation of ALNA conversion may limit the influence of variations in dietary n 3 fatty acid intake on plasma lipid compositions. PMID- 12908892 TI - Ontogeny and nutritional manipulation of mitochondrial protein abundance in adipose tissue and the lungs of postnatal sheep. AB - The present study examined the ontogeny of mitochondrial protein abundance in adipose tissue and lungs over the first month of life in the sheep and the extent to which this may be altered by maternal undernutrition during the final month of gestation. The ontogeny of uncoupling protein (UCP), voltage-dependent anion channel (VDAC) and cytochrome c abundance were determined in adipose tissue and lungs sampled from near-term fetuses and young sheep aged 4 h, 1, 7 and 30 d. In adipose tissue, the abundance of UCP1, VDAC and cytochrome c all peaked at 1 d of age and then decreased by 30 d of age, at which stage the brown adipose tissue specific UCP1 was no longer detectable but UCP2 was clearly abundant. For the lungs, however, UCP2 and VDAC abundance both peaked 7 d after birth and then decreased by 30 d of age. During postnatal development, therefore, a marked change in mitochondrial protein abundance occurs within both adipose tissue and lungs. Maternal nutrient restriction had no effect on lamb growth or tissue weights at 30 d of age but was associated with increased abundance of UCP2 and VDAC but not cytochrome c in both adipose tissue and lungs. These mitochondrial adaptations within both adipose tissue and the lungs of offspring born to previously nutrient-restricted mothers may compromise adipose tissue and lung function during periods of environmental stress. PMID- 12908893 TI - Effects of dietary fat quantity and composition on fasting and postprandial levels of coagulation factor VII and serum choline-containing phospholipids. AB - Dietary fat influences plasma levels of coagulation factor VII (FVII) and serum phospholipids (PL). It is, however, unknown if the fat-mediated changes in FVII are linked to PL. The present study aimed to investigate the effects of dietary fat on fasting and postprandial levels of activated FVII (FVIIa), FVII coagulant activity (FVIIc), FVII protein (FVIIag) and choline-containing PL (PC). In a randomized single-blinded crossover-designed study a high-fat diet (HSAFA), a low fat diet (LSAFA), both rich in saturated fatty acids, and a high-fat diet rich in unsaturated fatty acids (HUFA) were consumed for 3 weeks. Twenty-five healthy females, in which postprandial responses were studied in a subset of twelve, were included. The HSAFA diet resulted in higher levels of fasting FVIIa and PC compared with the LSAFA and the HUFA diets (all comparisons P< or =0.01). The fasting PC levels after the LSAFA diet were also higher than after the HUFA diet (P<0.001). Postprandial levels of FVIIa and PC were highest on the HSAFA diet and different from LSAFA and HUFA (all comparisons P< or =0.05). Postprandial FVIIa was higher on the HUFA compared with the LSAFA diet (P<0.03), whereas the HUFA diet resulted in lower postprandial levels of PC than the LSAFA diet (P<0.001). Significant correlations between fasting levels of PC and FVIIc were found on all diets, whereas FVIIag was correlated to PC on the HSAFA and HUFA diet. The present results indicate that dietary fat, both quality and quantity, influences fasting and postprandial levels of FVIIa and PC. Although significant associations between fasting FVII and PC levels were found, our results do not support the assumption that postprandial FVII activation is linked to serum PC. PMID- 12908894 TI - The effect of retinyl palmitate added to iron-fortified maize porridge on erythrocyte incorporation of iron in African children with vitamin A deficiency. AB - Retinyl palmitate added to Fe-fortified maize bread has been reported to enhance Fe absorption in adult Venezuelan subjects but not in Western Europeans. It is not known to what extent these results were influenced by differences in vitamin A status of the study subjects. The objective of the present study was to evaluate the influence of retinyl palmitate added to Fe-fortified maize porridge on erythrocyte incorporation of Fe in children with vitamin A deficiency, before and after vitamin A supplementation. Erythrocyte incorporation of Fe-stable isotopes was measured 14 d after intake of maize porridge (2.0 mg Fe added as ferrous sulfate) with and without added retinyl palmitate (3.5 micromol; 3300 IU). The study was repeated 3 weeks after vitamin A supplementation (intake of a single dose of 210 micromol retinyl palmitate; 'vitamin A capsule'). Vitamin A status was evaluated by the modified relative dose-response (MRDR) technique. Retinyl palmitate added to the test meal reduced the geometric mean erythrocyte incorporation of Fe at baseline from 4.0 to 2.6 % (P=0.008, n 13; paired t test). At 3 weeks after vitamin A supplementation, geometric mean erythrocyte incorporation was 1.9 and 2.3 % respectively from the test meal with and without added retinyl palmitate (P=0.283). Mean dehydroretinol:retinol molar ratios were 0.156 and 0.125 before and after intake of the single dose of 210 micromol retinyl palmitate; 'vitamin A capsule' (P=0.15). In conclusion, retinyl palmitate added to the labelled test meals significantly decreased erythrocyte incorporation of Fe in children with vitamin A deficiency at baseline but had no statistically significant effect 3 weeks after vitamin A supplementation. The difference in response to retinyl palmitate added to Fe-fortified maize porridge on erythrocyte incorporation of Fe before and after intake of the vitamin A capsule indicates, indirectly, changes in vitamin A status not measurable by the MRDR technique. The lack of conclusive data on the effect of retinyl palmitate on Fe absorption indicates the complexity of the interactions between vitamin A status, dietary vitamin A and Fe metabolism. PMID- 12908895 TI - Effect of reduced maternal protein intake in pregnancy in the rat on the fatty acid composition of brain, liver, plasma, heart and lung phospholipids of the offspring after weaning. AB - Reduced protein intake during pregnancy decreased maternal hepatic and plasma docosahexaenoic acid concentrations and impaired docosahexaenoic acid accumulation into fetal brain in the rat. The present study investigated whether restriction of maternal protein intake during pregnancy in the rat alters membrane phospholipid fatty acid composition in the offspring after weaning. Female rats (six per group) were mated and fed diets containing either 180 or 90 g protein/kg throughout pregnancy. Mothers were transferred to standard chow after delivery and the litters reduced to eight pups. Weaning was at 28 d and pups were killed 5 to 6 d later. Tissue weights or membrane total phosphatidylcholine (PC) and phosphatidylethanolamine (PE) concentrations in the offspring did not differ between dietary groups. There were significant differences between the 180 and 90 g/kg groups in liver, brain, lung and heart fatty acid composition that differed between tissues and phospholipid classes. For example, docosahexaenoic and arachidonic acid concentrations were 23 and 10 % lower respectively in hepatic PC, but not PE, in the 90 g/kg group. In brain, docosahexaenoic acid concentration was 17 % lower in PC, but not PE, while arachidonic acid content was 21 % greater in PE but unchanged in PC. The greatest differences were in unsaturated fatty acids, which suggests alterations to desaturase activities and/or the specificity of phospholipid biosynthesis. These results suggest that restricted maternal protein intake during pregnancy results in persistent alterations to membrane fatty acid content. PMID- 12908896 TI - Use of biomarkers of sub-clinical infection, nutrition and neonatal maturity to interpret plasma retinol in Nigerian neonates. AB - Using the World Health Organization criterion, the prevalence of sub-clinical vitamin A deficiency can be assessed using plasma retinol concentrations <0.7 micromol/l. However, plasma retinol can be depressed by infection; thus, the use of this criterion alone may overestimate deficiency. In the present study, we investigated the usefulness of the acute-phase proteins (APP) alpha1 antichymotrypsin (ACT) and alpha1-acid glycoprotein (AGP), plasma carotenoids and anthropometric and gestational indices to interpret plasma retinol in the blood of 192 apparently healthy Nigerian neonates collected randomly during days 1-20 postpartum. The mean weight (2.64 kg) and length (0.458 m) of the neonates and plasma concentrations (geometric mean, micromol/l) of retinol (0.54), alpha carotene (0.072), ss-carotene (0.076) and lutein (0.080) were low. The prevalence of vitamin A deficiency was 72 %, indicating a severe public health problem. Babies who were of low birth weight (P<0.003) or premature and low birth weight (P<0.023) had significantly lower retinol concentrations than full-term normal weight babies. Thirty-two neonates had abnormal ACT and forty-four abnormal AGP concentrations. Positive correlations between retinol and ACT (r 0.186, P=0.05) and AGP (r 0.31, P=0.0001) during days 1-5 may be due to the increasing plasma retinol from maternal milk and a coincidental increasing capacity to synthesise APP. Subsequently, negative correlations between retinol and ACT (r -0.28, P=0.02) and AGP (r -0.29, P=0.018) from day 6 onwards reflected the continuing increase in plasma retinol, but no further increase in the APP. Overall, weight, ACT, lutein and age explained 30 % of the variance in retinol, but lutein was the most significant (r(2) 0.18, P<0.0001). Hence, the distribution of plasma retinol concentrations in this group of neonates was more strongly linked with nutrition (via the surrogate marker lutein) than infection. PMID- 12908897 TI - Single oral doses of 13C forms of pteroylmonoglutamic acid and 5 formyltetrahydrofolic acid elicit differences in short-term kinetics of labelled and unlabelled folates in plasma: potential problems in interpretation of folate bioavailability studies. AB - Single (13)C6-labelled doses of pteroylmonoglutamic acid (PteGlu; 634 nmol) or 5 formyltetrahydrofolic acid (431-569 nmol) were given to fasted adult volunteers, and the rise in total and (13)C-labelled plasma 5-methyltetrahydrofolic acid metabolite monitored over 8 h by HPLC and liquid chromatography-MS. The dose adjusted area under the curve (AUC) for total (labelled plus unlabelled) plasma 5 methyltetrahydrofolic acid following a 5-formyltetrahydrofolic acid test dose was 155 % that obtained following a PteGlu test dose. Surprisingly, an average 60 and 40 % of the total plasma 5-methyltetrahydrofolic acid response to [(13)C6]PteGlu and [(13)C6]5-formyltetrahydrofolic acid, respectively, was unlabelled; an observation never before reported. Short-term kinetics of plasma [(13)C6]5 methyltetrahydrofolic acid showed a slower initial rate of increase in plasma concentration and longer time to peak following an oral dose of [(13)C6]PteGlu compared with that for an oral dose of [(13)C6]5-formyltetrahydrofolic acid, while the [(13)C6]5-methyltetrahydrofolic acid AUC for [(13)C6]5 formyltetrahydrofolic acid was 221 % that for [(13)C6]PteGlu. These data indicate that PteGlu and 5-formyltetrahydrofolic acid, which are thought to be well absorbed (about 90 %) at physiological doses, exhibit dramatically different rates and patterns of plasma response. A limitation in the rate of reduction of PteGlu before methylation could result in slower mucosal transfer of [(13)C6]5 methyltetrahydrofolic acid derived from [(13)C6]PteGlu into the plasma. This, when coupled with an observed similar plasma clearance rate for [(13)C6]5 methyltetrahydrofolic acid metabolite derived from either folate test dose, would yield a comparatively smaller AUC. These findings suggest potential problems in interpretation of absorption studies using unlabelled or labelled folates where the rate of increase, the maximum increase, or the AUC, of plasma folate is employed for test foods (mainly reduced folates) v. a 'reference dose' of PteGlu. PMID- 12908898 TI - The rat as a model for pigs: comparative values for the digestibility of NSP and other macronutrients. AB - The present investigation was undertaken to study whether conventional male Wistar rats could be used as a model for pigs with regard to total tract digestibilities of NSP and macronutrients and whether nebacitin-treated rats could be used as a model for small intestinal digestibility in pigs. Nineteen experimental diets prepared from different fractions of wheat and oats, and which all had been evaluated in experiments with ileal cannulated pigs, were used for the present study. There was a close relationship between the total tract digestibilities of organic matter in the two species. The same was the case with regard to the digestibility of total NSP and arabinoxylans, but the values were on average 6 % lower in rats than in pigs. On average, there were no significant differences between rats and pigs with regard to faecal protein digestibility. However, protein in oat-based diets was significantly better digested in the rat than the pig. The digestibility of fat was consistently higher in rats than in pigs, with the biggest difference being found in oat-based diets, in which most of the fat was locked in cell structures. For the wheat-based diets, in which a large proportion of the fat was present as added fat, there was a greater similarity between the two species. In nebacitin-treated rats the digestibility of organic matter, starch, protein and fat was negatively related to the dietary level of NSP, but this model could not be used to predict the small intestinal digestibility of NSP and macronutrients in ileal-cannulated pigs. PMID- 12908899 TI - Dietary docosahexaenoic acid-induced production of tissue lipid peroxides is not suppressed by higher intake of ascorbic acid in genetically scorbutic Osteogenic Disorder Shionogi/Shi-od/od rats. AB - In previous studies, we showed that docosahexaenoic acid (DHA) ingestion enhanced the susceptibility of rat liver and kidney to lipid peroxidation, but did not increase lipid peroxide formation to the level expected from the relative peroxidizability index (P-index) of the total tissue lipids. The results suggested the existence of some suppressive mechanisms against DHA-induced tissue lipid peroxide formation, as increased tissue ascorbic acid (AsA) and glutathione levels were observed. Therefore, we focused initially on the role of AsA for the suppressive mechanisms. For this purpose, we examined the influence of different levels of dietary AsA (low, moderate, high and excessive levels were 100, 300 (control), 600 and 3000 mg/kg diet respectively) on the tissue lipid peroxide and antioxidant levels in AsA-requiring Osteogenic Disorder Shionogi/Shi-od/od (ODS) rats fed DHA (6.4 % total energy) for 32 or 33 d. Diets were pair-fed to the DHA- and 100 mg AsA/kg diet-fed group. We found that the lipid peroxide concentrations of liver and kidney in the DHA-fed group receiving 100 mg AsA/kg diet were significantly higher or tended to be higher than those of the DHA-fed groups with AsA at more than the usual control level of 300 mg/kg diet. Contrary to this, the liver alpha-tocopherol concentration was significantly lower or tended to be lower in the DHA and 100 mg AsA/kg diet-fed group than those of the other DHA-fed groups. However, tissue lipid peroxide formation and alpha-tocopherol consumption were not suppressed further, even after animals received higher doses of AsA. The present results suggest that higher than normal concentrations of tissue AsA are not necessarily associated with the suppressive mechanisms against dietary DHA induced tissue lipid peroxide formation. PMID- 12908900 TI - Influence of plant and bacterial myrosinase activity on the metabolic fate of glucosinolates in gnotobiotic rats. AB - The breakdown of glucosinolates, a group of thioglucoside compounds found in cruciferous plants, is catalysed by dietary or microbial myrosinase. This hydrolysis releases a range of breakdown products among which are the isothiocyanates, which have been implicated in the cancer-protective effects of cruciferous vegetables. The respective involvement of plant myrosinase and gut bacterial myrosinase in the conversion, in vivo, of glucosinolates into isothiocyanates was investigated in sixteen Fischer 344 rats. Glucosinolate hydrolysis in gnotobiotic rats harbouring a whole human faecal flora (Flora+) was compared with that in germ-free rats (Flora-). Rats were offered a diet where plant myrosinase was either active (Myro+) or inactive (Myro-). The conversion of prop-2-enyl glucosinolate and benzyl glucosinolate to their related isothiocyanates, allyl isothiocyanate and benzyl isothiocyanate, was estimated using urinary mercapturic acids, which are endproducts of isothiocyanate metabolism. The highest excretion of urinary mercapturic acids was found when only plant myrosinase was active (Flora-, Myro+ treatment). Lower excretion was observed when both plant and microbial myrosinases were active (Flora+, Myro+ treatment). Excretion of urinary mercapturic acids when only microbial myrosinase was active (Flora+, Myro- treatment) was low and comparable with the levels in the absence of myrosinase (Flora-, Myro- treatment). No intact glucosinolates were detected in the faeces of rats from the Flora+ treatments confirming the strong capacity of the microflora to break down glucosinolates. The results confirm that plant myrosinase can catalyse substantial release of isothiocyanates in vivo. The results also suggest that the human microflora may, in some circumstances, reduce the proportion of isothiocyanates available for intestinal absorption. PMID- 12908901 TI - Inhibition of tumour necrosis factor-alpha and interleukin 6 production by mononuclear cells following dietary fish-oil supplementation in healthy men and response to antioxidant co-supplementation. AB - Increased dietary consumption of the n-3 polyunsaturated fatty acids (PUFA) eicosapentaenoic acid (20 : 5n-3; EPA) and docosahexaenoic acid (22 : 6n-6; DHA) is associated with their incorporation into circulating phospholipid and increased production of lipid peroxide metabolites. The relationship between peripheral blood mononuclear cell (PBMC) function, n-3 PUFA intake and antioxidant co-supplementation is poorly defined. We therefore investigated tumour necrosis factor (TNF)-alpha and interleukin (IL) 6 production by PBMC and phospholipid fatty acid composition in plasma and erythrocytes of healthy male subjects (n 16) receiving supplemental intakes of 0.3, 1.0 and 2.0 g EPA+DHA/d, as consecutive 4-week courses. All subjects were randomised in a double-blind manner to receive a concurrent antioxidant supplement (200 microg Se, 3 mg Mn, 30 mg D-alpha-tocopheryl succinate, 90 mg ascorbic acid, 450 microg vitamin A (beta carotene and retinol)) or placebo. There was a positive dose-dependent relationship between dietary n-3 PUFA intake and EPA and DHA incorporation into plasma phosphatidylcholine and erythrocyte phosphatidylethanolamine, with a tendency towards a plateau at higher levels of intake. Production of TNF-alpha and IL-6 by PBMC decreased with increasing n-3 PUFA intake but tended towards a 'U-shaped' dose response. Both responses appeared to be augmented by antioxidant co-supplementation at intermediate supplementary n-3 PUFA intakes. Thus, increased dietary n-3 PUFA consumption resulted in defined but contrasting patterns of modulation of phospholipid fatty acid composition and PBMC function, which were further influenced by antioxidant intake. PMID- 12908902 TI - Effect of isoenergetic low- and high-carbohydrate diets on substrate kinetics and oxidation in healthy men. AB - The effect of diet composition on post-absorptive (15 h fast) fatty acid and glucose metabolism was investigated in five healthy men after 2 weeks on a low carbohydrate (Low-CHO) diet (30 % energy intake from carbohydrates, 55 % from fat, 15 % from protein) and after 2 weeks on a high-carbohydrate (High-CHO) diet (energy intake 75, 10 and 15 % from carbohydrates, fat and protein respectively). The diets were isoenergetic and comprised real foods. Stable-isotope tracer methodology and indirect calorimetry were employed to measure glucose and fatty acid kinetics and oxidation. The relative contribution of carbohydrate to the total energy expenditure was significantly higher after the High-CHO diet. After the High-CHO diet, total and plasma fatty oxidation (2.4 (SE 0.7) and 2.1 (SE 0.4) micromol/kg per min respectively) were significantly lower than after the Low-CHO diet (4.8 (SE 0.5) and 4.6 (SE 0.8) micromol/kg per min for total and plasma fatty oxidation respectively). The rate of appearance (Ra) of non esterified fatty acids (NEFA) in plasma and the arterial NEFA concentration were both significantly lower following the High-CHO than the Low-CHO diet. However, even after the High-CHO diet, NEFA Ra was threefold higher than plasma fatty acid oxidation. Thus, the decrease in fatty acid oxidation after consumption of a high carbohydrate diet for 2 weeks in healthy men is unlikely to result from decreased fatty acid delivery to the tissues. Glucose Ra and arterial plasma glucose concentration were similar after the two diets. After the High-CHO diet, arterial lactate concentration was higher and total carbohydrate oxidation rate well exceeded glucose Ra in plasma. Therefore, alterations in intracellular mechanisms may limit fatty acid oxidation after high-carbohydrate diets. PMID- 12908903 TI - Intra-individual variation of basal metabolic rate and the influence of daily habitual physical activity before testing. AB - The present study determined the intra-individual variation of BMR measurements, using a standard out-patient protocol, with the subjects transporting themselves to the laboratory for the BMR measurements after spending the night at home. The effect of a non-fasting state and variation in daily habitual physical activity the day before testing was evaluated. Eight male and eleven female subjects participated in three BMR measurements with 2-week intervals. Physical activity was estimated with a tri-axial accelerometer for movement registration, during the 3 d before each BMR measurement. There were no significant differences in estimated BMR (ANOVA repeated measures, P=0.88) and in physical activity (ANOVA repeated measures, P=0.21). Mean within-subject CV in BMR was found to be 3.3 (SD 2.1) %, ranging from 0.4 to 7.2 %. Differences between BMR measurements could not be explained by differences in physical activity the day before; however the mean within-subject CV in BMR changed from 5.7 to 5.2 % after correcting for within machine variability and from 5.2 to 3.3 % after excluding five measurements because of non-compliance to the protocol including fasting. In conclusion, BMR values measured with a standard out-patient protocol are sufficiently reproducible for most practical purposes despite the within-subject variability in physical activity the day before the measurement. For this purpose, however, non-fasting subjects must be excluded and a regular function check of the ventilated-hood system is recommendable. PMID- 12908904 TI - Can prescription of sip-feed supplements increase energy intake in hospitalised older people with medical problems? AB - A blinded randomised controlled trial of prescribed oral sip-feed supplements compared with routine hospital practice was undertaken in acute admissions to a geriatric medicine department. Patients were eligible for inclusion if they were admitted from home, were not obese (BMI>75th percentile), had no swallowing difficulties and were not deemed to be in the terminal stage of illness. On admission they were stratified by nutritional status (BMI<5th, >5th to <25th, >25th to <75th percentile) and randomised. The intervention group received 120 ml oral sip-feed supplement prescribed three times per d in the medicine prescription chart (22.5 g protein, 2260 kJ (540 kcal) energy/d) distributed at medication rounds for the duration of hospital stay. The control group received routine hospital care. Outcomes were patient compliance with supplement, total energy intake and nursing staff views of the method. Patients were randomised to receive supplements (n 186 of total n 381). Half had full compliance and three quarters at least moderate compliance. Total energy intake was significantly increased, on average, in the intervention group (P=0.001). The proportion of patients meeting estimated minimum energy requirements was significantly increased (P=0.023), but was still <50 % for the sample of patients in the intervention group. The present study suggests this method is acceptable to patients and staff and improves total energy intake. However, the amount prescribed did not ensure minimum energy requirements were met in all cases. PMID- 12908905 TI - Relationship between body mass index, serum cholesterol, leisure-time physical activity, and diet in a Mediterranean Southern-Europe population. AB - The aim of the present study was to determine the relationship of BMI with other cardiovascular risk factors, leisure-time physical activity and diet. Participants were recruited in a cross-sectional population-based survey in a southern-Europe Mediterranean population (Spain); cardiovascular risk factors were measured, and leisure-time physical activity and diet intake were evaluated. Linear regression analysis adjusted for several confounders showed a significant, direct association of BMI and total cholesterol (P<0.005) and LDL-cholesterol (P<0.006), in men. HDL-cholesterol was inversely related to BMI in both sexes (P<0.0001). Higher BMI was more frequent in less-active men (P<0.04) but not in women. BMI increased significantly (P<0.0001) by 1.92 kg/m(2) with each 4.18 MJ consumed in men but not in women. Dietary intakes of carbohydrate (P<0.03), total fat (P<0.03) and saturated fatty acids (P<0.02) were directly associated with BMI in men but not in women, in whom protein intake was correlated (P<0.001) with BMI. Linear regression models including dietary components explained up to 10.6 and 21.1 % of BMI variability in men and women, respectively. Sex differences in the association of BMI with total cholesterol, and LDL-cholesterol, may account for the lower risk for CHD in women compared with men of similar BMI reported in the literature for the southern-Europe Mediterranean region. An increases of BMI may be more deleterious in populations in which it is accompanied by other risk factors such as a higher intake of total fat and, particularly, of saturated fatty acids, or lower leisure-time physical activity. PMID- 12908906 TI - Associations between intake of added sugars and intakes of nutrients and food groups in the diets of German children and adolescents. AB - The aim of the present analysis was to evaluate the effect of intake of added sugars on intakes of nutrients and food groups. We used a mixed linear model (PROC MIXED) to analyse data from the Dortmund Nutritional and Anthropometric Longitudinally Designed (DONALD) study (an ongoing longitudinal cohort study between infancy and adulthood). Between July 1985 and August 2001, 3 d weighed dietary records (n 4993) from 849 children and adolescents (416 male, 433 female) aged 2-18 years were collected and evaluated. Overall mean intake of added sugars was 12.4 (SD 5.3) (median 12.0) % energy. Fat intake (% energy) and intake of all analysed nutrients (as % reference values for nutrient intakes) fell across quintiles 1-5 of energy intake from added sugars (P<0.001). With the exception of 'Sugary foods' and 'Beverages' (significant increase, P<0.0001) and 'Dairy' (NS), intake of all other food groups decreased with increasing intake of added sugars (P<0.0001). In conclusion, we found a slight, but statistically significant, nutrient-dilution effect of added sugars and a significant reduction in intake of important nutrient-bearing food groups. However, the DONALD study gives no clear indicator on grounds of micronutrient dilution for a quantitative limit on intake of added sugars. PMID- 12908907 TI - Antioxidative probiotic fermented goats' milk decreases oxidative stress-mediated atherogenicity in human subjects. AB - The increasing interest in a healthy diet is stimulating innovative development of novel scientific products in the food industry. The viable lactic acid bacteria in fermented milk products, such as yoghurt, have been associated with increased lactose tolerance, a well-balanced intestinal microflora, antimicrobial activity, stimulation of the immune system and antitumoural, anticholesterolaemic and antioxidative properties in human subjects. Recently, we have studied a human Lactobacillus spp. strain that possesses antioxidative activity. The aim of the present pilot study was to develop goats' milk fermented with the human antioxidative lactobacilli strain, Lactobacillus fermentum ME-3, and to test the effect of the fermented probiotic goats' milk on oxidative stress markers (including markers for atherosclerosis) in human blood and urine and on the gut microflora. Twenty-one healthy subjects were assigned to two treatment groups: goats' milk group and fermented goats' milk group (150 g/d) for a period of 21 d. Consumption of fermented goats' milk improved anti-atherogenicity in healthy subjects: it prolonged resistance of the lipoprotein fraction to oxidation, lowered levels of peroxidized lipoproteins, oxidized LDL, 8-isoprostanes and glutathione redox ratio, and enhanced total antioxidative activity. The consumption of fermented goats' milk also altered both the prevalence and proportion of lactic acid bacteria species in the gut microflora of the subjects. We conclude that the goats' milk fermented with our special antioxidative lactobacilli strain Lactobacillus fermentum ME-3 exhibits anti-atherogenic effects. PMID- 12908908 TI - A potential protective mechanism of soya isoflavones against 7,12 dimethylbenz[a]anthracene tumour initiation. AB - Epidemiological studies indicate that Asian women have a lower breast cancer incidence compared with their counterparts in the West, and the difference has been related to soya consumption. Animal studies have suggested that soya may prevent dimethylbenz[a]anthracene (DMBA)-induced carcinogenesis in the breast. In the present study a cell culture model was developed to address the effect of soya isoflavones on the DMBA-induced DNA damage. DMBA is metabolized into a DNA attacking moiety by two phase I cytochrome P450 (CYP) enzymes CYP1A1 and CYP1B1. DNA mutation caused by this genotoxic agent is a crucial step in cancer initiation. Substances that interfere with the CYP1 enzyme activities can affect the initiation. In the present study, genistein was found to be an effective inhibitor of recombinant human CYP1A1 and CYP1B1 with Ki of 15.35 and 0.68 micromol/l. The other soya isoflavone daidzein, on the other hand, did not demonstrate any significant inhibition of the enzyme activities. At the transcriptional level, DMBA induced the CYP1 enzyme expressions by stimulating the xenobiotic response element (XRE)-dependent transactivation pathway. When genistein (25 micromol/l) was co-administered with DMBA, the XRE-Luc activity the CYP1 mRNA abundances were significantly suppressed. The present study illustrated that the soya isoflavone genistein, but not daidzein, protected against DMBA genotoxicity. PMID- 12908909 TI - New aspects in pathogenesis of konzo: neural cell damage directly caused by linamarin contained in cassava (Manihot esculenta Crantz). AB - Epidemic spastic paraparesis (konzo) found in tropical and subtropical countries is known to be caused by long-term intake of cassava (Manihot esculenta Crantz), which contains a cyanoglucoside linamarin (alpha-hydroxyisobutyronitrile-beta-d glucopyranoside). It has been reported that linamarin is enzymatically converted to cyanide by bacteria in the intestine, and this is absorbed into the blood and then damages neural cells. However, unmetabolized linamarin was found in the urine after oral administration of cassava; thus, we hypothesized that konzo could be caused by direct toxicity of the unmetabolized linamarin that was transferred to the brain and could be transported into neural cells via a glucose transporter. In the present study it was confirmed that linamarin directly damaged neural culture pheochromocytoma cell (PC) 12 cells; 0.10 mm-linamarin caused cell death at 13.31 (SD 2.07) %, which was significantly different from that of control group (3.18 (SD 0.92) %, P=0.0004). Additional 10 microM cytochalasin B, an inhibitor of a glucose transporter, prevented cell death: the percentage of dead cells significantly decreased to 6.06 (SD 1.98), P=0.0088). Furthermore, glucose also prevented cell death. These present results strongly suggest that linamarin competes with cytochalasin B and glucose for binding to a glucose transporter and enters into cells via glucose transporter. PMID- 12908910 TI - Folate bioavailability: UK Food Standards Agency workshop report. AB - The UK Food Standards Agency convened a group of expert scientists to review current research investigating folate bioavailability. The workshop aimed to overview current research and establish priorities for future research. Discrepancies were observed in the evidence base for folate bioavailability, especially with regard to the relative bioavailability of natural folates compared with folic acid. A substantial body of evidence shows folic acid to have superior bioavailability relative to food folates; however, the exact relative bioavailability still needs to be determined, and in particular with regard to mixed diets. The bioavailability of folate in a mixed diet is probably not a weighted average of that in the various foods consumed; thus the workshop considered that assessment of folate bioavailability of whole diets should be a high priority for future research. PMID- 12908911 TI - National Academies of Science definitions relating to food fibre only add confusion. PMID- 12908912 TI - The life cycle of Reticulitermes spp (Isoptera: Rhinotermitidae): what do we know? AB - The subterranean termites in the genus Reticulitermes have a complex and plastic life cycle, which has been the subject of a number of publications over the past century. Given the inherent difficulties in studying such cryptic, eusocial organisms it is not perhaps surprising that the literature on their biology has failed to reach a consensus. An overview of the literature is given, which is followed by a discussion of the various theories on the life cycle of Reticulitermes spp. A substantial proportion of the review focuses on the French literature, which constitutes the majority of the primary sources and can be difficult to access. There are many discrepancies in the literature in terms of the number of instars, the definition of workers and the question of whether they should be termed pseudergates or, potentially, an additional terminology used to differentiate between pseudergates and the true workers seen in the higher termites (Isoptera: Termitidae). It remains very difficult to compare publications as there is little conformity; a problem that is aggravated by a general absence of drawings of the relevant instars. Further work on the biology of Reticulitermes is clearly required. There is also a need for researchers to agree on a standard terminology for this genus. A glossary is provided for the various synonyms and definitions. PMID- 12908913 TI - Localized breeding of the Anopheles gambiae complex (Diptera: Culicidae) along the River Gambia, West Africa. AB - A study was undertaken to identify the major larval habitats of the Anopheles gambiae (Giles) complex in rural Gambia. Mosquito larvae and pupae were sampled along transects and in specific habitats in the central region of the country during the rainy seasons of 1996 and 1997. The sampling showed that the major breeding sites were located on the flooded alluvial soils bordering the river. The largest numbers of larvae were found during September, one month after the peak rains. Polymerase chain reaction analysis of specimens showed that Anopheles melas (Theobald) was the dominant species in the flooded areas (81.5%), followed by A. gambiae sensu stricto (Giles) (18.0%) and A. arabiensis (Patton) (0.5%). By sampling in specific habitats it was evident that A. arabiensis was mainly breeding in rain-fed rice fields along the edge of the alluvial soils. Anopheles melas and A. gambiae s.s. often coexisted but whereas A. melas were found in water with a salinity of up to 72% sea water (25.2 g NaCl l(-1)), A. gambiae s.s. only occurred in water with up to 30% sea water (10.5 g NaCl l(-1)). Anopheles melas larvae were found in association with plant communities dominated by sedges and grasses (Eleocharis sp., Paspalum sp., Sporobolus sp.) and sea-purslane Sesuvium portulacastrum (L.) and the presence of cattle hoof prints, whereas A. gambiae s.s. larvae mainly occurred in association with Paspalum sp. and Eleocharis sp. The study showed that even during the peak rainy season, breeding of the A. gambiae complex is almost entirely restricted to the extensive alluvial areas along the river. PMID- 12908914 TI - Incidence of insecticide resistance alleles in sexually-reproducing populations of the peach-potato aphid Myzus persicae (Hemiptera: Aphididae) from southern France. AB - Intensive chemical treatments have led to the development of a number of insecticide resistance mechanisms in the peach-potato aphid Myzus persicae (Sulzer). Some of these mechanisms are known to be associated with negative pleiotropic effects (resistance costs). Molecular and biochemical methods were used to determine the genotypes or phenotypes associated with four insecticide resistance mechanisms in single aphids from sexually-reproducing populations in southern France. The mechanisms considered were E4 and FE4 carboxylesterase overproduction, modified acetycholinesterase, and kdr and rdl resistance associated mutations. A new method for determining individual kdr genotypes is presented. Almost all resistant individuals overproduced FE4 carboxylesterase, whereas modified acetylcholinesterase was rare. Both the kdr and rdl resistance mutations were present at high frequencies in French sexually-reproducing populations. The frequencies of insecticide resistance genes were compared before and after sexual reproduction in one peach orchard at Avignon to evaluate the potential impact of selection on the persistence of resistance alleles in the over-wintering phase. The frequencies of the kdr and rdl mutations varied significantly between autumn and spring sampling periods. The frequency of the kdr mutation increased, probably due to pyrethroid treatments at the end of the winter. Conversely, the frequency of the rdl mutation decreased significantly during winter, probably because of a fitness cost associated with this mutation. PMID- 12908915 TI - Biological control of the larger grain borer Prostephanus truncatus (Coleoptera: Bostrichidae) in Kenya using a predatory beetle Teretrius nigrescens(Coleoptera: Histeridae). AB - The establishment of the predatory histerid beetle, Teretrius nigrescens Lewis, following its field release in south eastern Kenya in 1992, and its impact on populations of the larger grain borer, Prostephanus truncatus (Horn), in woodland habitats is evaluated. Pheromone trap catch data show a single strong peak of P. truncatus from November to January, coincident with the short rains. Comparisons of P. truncatus pheromone trap catches before and after predator establishment, and in areas with and without T. nigrescens, show a sustained decline of over 80% in P. truncatus abundance over a 5-year period, following the establishment T. nigrescens. Concurrent laboratory studies of the impact of T. nigrescens on populations of P. truncatus raised on a wood substrate showed a 77% reduction in the growth of the pest population. These results are compared with published findings of the impact of T. nigrescens as a biological control agent of P. truncatus in west Africa. PMID- 12908916 TI - Overwintering, soil distribution and phenology of Childers canegrub, Antitrogus parvulus (Coleoptera: Scarabaeidae) in Queensland sugarcane. AB - In this study, the question of whether Childers canegrub, Antitrogus parvulus (Britton) overwinters in the subsoil was addressed. Irrigated fields of sugarcane were sampled during a 2-year period near Bundaberg in southern Queensland. Antitrogus parvulus overwintered as second and third instars at each of three sites. During autumn and winter third instars of different allochronic (separated in age by 12 months) populations occurred together and could not be readily separated. Field-collected third instars were reared on ryegrass and separated into two age groups based on the date of pupation. Third instars in the first year of their life cycle (young third instars) remained at shallow depth (100-200 mm) and did not overwinter in the subsoil as once thought. Minimum temperatures during winter were 13-16 degrees C and did not prevent young third instars from feeding and gaining weight. Third instars in their second and final year moved downwards from late summer and pupated in the subsoil at 293-425 mm in spring. General phenology was as previously reported with first instar larvae occurring from January until April, second instars from January until November and third instar larvae throughout the year. Prepupae and pupae were found between October and December and adults occurred in soil during November and January. Batches of eggs occurred at a mean depth of 350 mm. First and second instars occurred predominantly at relatively shallow (100-200 mm) depths in the soil profile. All stages tended to be most common under rows of sugarcane rather than in the interrow. PMID- 12908917 TI - Navigation of Lutzomyia longipalpis (Diptera: Psychodidae) under dusk or starlight conditions. AB - The responses of male and female Lutzomyia longipalpis (Lutz & Neiva) to different wavelengths of light was tested by presenting the sandflies with two light sources simultaneously, a series of test wavelengths between 350-670 nm and a 400 nm control. To test whether L. longipalpis could discriminate between the test and control, three sets of experiments were carried out in which the test wavelengths were presented at higher, equivalent or lower intensity than the control. In all three experiments, ultra-violet (350 nm) and blue-green-yellow (490-546 nm) light was more attractive to L. longipalpis than the control wavelength. However, at low intensity, UV was less attractive, than equivalent or higher intensity UV light. At intensities equivalent to or higher than the control wavelength, ultra-violet light was more attractive than blue-green. Furthermore, at low intensity, green-yellow (546 nm) light was more attractive to males whereas blue-green (490 nm) was more attractive to females. Blue-violet (400 nm) and orange-red (600-670 nm) light were least attractive in all three sets of experiments. Response function experiments indicated that the responses were dependent on both intensity and wavelength and that therefore more than one photoreceptor must be involved in the response. The results indicated that L. longipalpis can discriminate between different wavelengths at different intensities and thus have true colour vision. It also suggests that L. longipalpis may be able to navigate at dusk or under moonlight or starlight conditions using light in the blue-green-yellow part of the spectrum. The difference in response of males and females to light in this region is interesting and may indicate the different ecology of the sexes at night. Overall, these results may have important implications for sandfly trap design. PMID- 12908918 TI - Patterns of resistance to three cereal aphids among wheats in the genus Triticum (Poaceae). AB - Forty-one accessions of wild and cultivated wheats belonging to 19 Triticum species were tested in the field for resistance to three species of aphids, Rhopalosiphum padi Linnaeus, Sitobion avenae Fabricius and Schizaphis graminum Rondani. Antibiotic resistance was estimated by the increase in biomass of aphids over 21 days on adult plants. Overall resistance was estimated by the plant biomass lost due to aphid infestation. All three species of aphids survived and reproduced on all wheats, and reduced spike biomass compared to uninfested controls. The level of antibiosis varied among wheat species and among accessions, with accessions from three, five and one species showing antibiosis to R. padi, S. avenae and S. graminum, respectively. Overall resistance to the three aphid species was observed in five to seven accessions per aphid species. Resistance was usually specific to one aphid species. The frequency of accessions with antibiosis or overall resistance was associated with the ploidy level of the plant species. Except for overall resistance to R. padi, resistance was highest for diploid species and lowest for hexaploid species. No consistent relationship between resistance and level of domestication was detected. Accessions of the wild wheats, Triticum boeoticum Bois, Triticum tauschii (Coss.) Schmal. and Triticum araraticum Jakubz. exhibited high levels of resistance to aphids, as did Triticum monococcum L. which is derived from T. boeoticum. Nevertheless, individual susceptible or resistant accessions occurred at all levels within the evolutionary tree of wheat. PMID- 12908919 TI - Response of female Cydia molesta (Lepidoptera: Tortricidae) to plant derived volatiles. AB - Peach shoot volatiles were attractive to mated female oriental fruit moth, Cydia molesta (Busck), in a dual choice arena. No preference was observed between leaf odours from the principle host plant, peach, and the secondary host plant, apple. Twenty-two compounds were identified in headspace volatiles of peach shoots using gas chromatography-mass spectrometry. Green leaf volatiles accounted for more than 50% of the total emitted volatiles. A bioassay-assisted fractionation using different sorbent polymers indicated an attractant effect of compounds with a chain length of 6-8 carbon atoms. The major compounds of this fraction were tested either singly or in combinations for behavioural response of females. Significant bioactivity was found for a three-component mixture of (Z)-3-hexen-1 yl acetate, (Z)-3-hexen-1-ol and benzaldehyde in a 4:1:1 ratio. This synthetic mixture elicited a similar attractant effect as the full natural blend from peach shoots as well as the bioactive fraction. PMID- 12908920 TI - The effect of chrysanthemum leaf trichome density and prey spatial distribution on predation of Tetranychus urticae (Acari: Tetranychidae) by Phytoseiulus persimilis (Acari: Phytoseiidae). AB - The effect of plant architecture, in terms of leaf hairiness, and prey spatial arrangement, on predation rate of eggs of the spider mite, Tetranychus urticae Koch, by the predatory mite Phytoseiulus persimilis Athias-Henriot was examined on cut stems of chrysanthemums. Three levels of leaf hairiness (trichome density) were obtained using two different chrysanthemum cultivars and two ages within one of the cultivars. The number of prey consumed by P. persimilis was inversely related to trichome density. At low prey densities (less than ten eggs per stem), prey consumption did not differ in a biologically meaningful way between treatments. The effect of prey spatial arrangement on the predation rate of P. persimilis was also examined. Predation rates were higher in prey patches on leaves adjacent to the release point of P. persimilis, but significantly greater numbers of prey were consumed in higher density prey patches compared to low density patches. The predators exhibited non-random searching behaviour, spending more time on leaves closest to the release point. The implications of these findings for biological control and predator-prey dynamics are discussed. PMID- 12908921 TI - Genetic delineation of sibling species of the pest fruit fly Bactocera (Diptera: Tephritidae) using microsatellites. AB - Using a large set of microsatellites, the genetic relationships between three closely related Australian fruit fly species, Bactrocera tryoni (Froggatt), B. neohumeralis (Hardy) and B. aquilonis(May) were investigated. Bactrocera tryoni and B. neohumeralis are sympatric, while B. aquilonisis allopatric to both. The sympatric species, B. tryoni and B. neohumeralis, were found to be genetically distinct. It is likely that despite differences in mating time between these two species, some gene flow still occurs. In contrast, the sibling species B. tryoni and B. aquilonis were found to be closely related, despite allopatry. The level of genetic divergence was similar to that found within eastern Australian populations of B. tryoni. Consideration of all available genetic data suggests that this similarity is not due to recent (i.e. within the last 30 years) displacement of B. aquilonis by B. tryoni from the B. aquilonis region (north western Australia). Instead the data suggests that, at least in the areas sampled, asymmetrical hybridization may have occurred over a longer timescale. PMID- 12908922 TI - Quantitative description and discrimination of butterfly wing patterns using moment invariant analysis. AB - Studies examining and using pattern variation in insects for identification and characterization of individuals and populations have been limited by the methods available for quantifying wing patterns objectively. In this paper, differences in wing pattern are demonstrated statistically using moment invariant data sets generated automatically from digitized images of the speckled wood butterfly, Pararge aegeria (Linnaeus). Studies with other biological subjects have already shown moment invariants to work well with outline shapes and silhouettes. A pilot study with replicated monochrome photographs of a single butterfly showed the method could detect pattern differences between wing surfaces, even in the presence of simulated wing fading and damage. In a further study of the wings of 228 specimens, multivariate analyses of variance using the moment data reliably detected differences between groups of butterflies according to sex, geographical origin and culture history. Potential applications and future improvements of the moment methodology are considered. PMID- 12908923 TI - A new rat model of diffuse brain injury associated with acute subdural hematoma: assessment of varying hematoma volume, insult severity, and the presence of hypoxemia. AB - The aim of this study was to develop a new rat model of diffuse brain injury (DBI) associated with acute subdural hemorrhage (SDH). In order to make this model more clinically relevant, we determined whether the varying hematoma volume, severity of DBI, or the presence of hypoxemia could influence the physiological consequence. SDH was made by an autologous blood injection, while DBI was induced using the impact acceleration model (mild, 450 g/1 m, severe, 450 g/2 m). Physiological parameters measured included intracranial pressure (ICP), mean arterial blood pressure (MABP), cerebral blood flow (CBF), and brain tissue water content. In the first series, 23 rats were randomized into the five following groups: Group 1, sham; Group 2, 400 (microL SDH; Group 3, SDH400 + mild DBI; Group 4, SDH400 + severe DBI; and Group 5, SDH300 + severe DBI. Results suggested that SDH300 + severe DBI (Group 5) may be the most suitable model, in which the MABP and CBF temporarily decreased during the SDH induction, but thereafter recovered to the baseline. Conversely, ICP was persistently elevated throughout the experiment. The water content was also significantly higher in both hemispheres compared to that of sham. In the second series, the animal was exposed to a hypoxemic insult (10 or 30 min) in addition to SDH300 + severe DBI (Group 6). The prolonged hypoxemia caused both a severe CBF reduction without recovery and a bilateral brain swelling, whereas the brief hypoxemia showed a gradual CBF recovery from the transient reduction and an increased water content only in the SDH side. These results suggest that these models may be potentially useful to study the combination of DBI and SDH with or without hypoxemia. PMID- 12908924 TI - Neurogenesis and glial proliferation persist for at least one year in the subventricular zone following brain trauma in rats. AB - In several models of traumatic brain injury in rodents, remarkably progressive tissue loss and neuron death has been observed accompanied by expanding ventricles. Here, we explored potential cell proliferation in the subventricular zone (SVZ) in response to this progressive posttraumatic neurodegeneration. Four month-old rats (n = 57) were subjected to parasagittal fluid-percussion brain injury or sham treatment (no injury), and their brains were harvested at 2 weeks, 2 months, 6 months, and 1 year (n = 6-8/group) after injury or sham treatment. Brain sections (6 microm) were stained with markers of cell proliferation, Ki-67, and proliferative cell nuclear antigen (PCNA) to detect mitotically active cells. In sham animals, we found a typical age-dependent decrease in Ki-67- and PCNA labelled cells in the SVZ over the course of 1 year. However, in brain-injured animals, this decrease was reversed culminating in a sixfold increase in the number of cells staining with Ki-67 and a threefold increase in cells staining with PCNA by 1 year following injury compared to age-matched controls. Using double labeling, we also determined that many of the cells staining with Ki-67 or PCNA expressed markers selective for neurons (neurofilament protein) and glia (GFAP). These data suggest that there is a persistent proliferation of neurons and glia in the SVZ following brain trauma that does not diminish during aging as found in non-injured animals. PMID- 12908925 TI - Anti-porin antibodies prevent excitotoxic and ischemic damage to brain tissue. AB - The mitochondrial permeability transition (MPT) is a converging event for different molecular routes leading to cellular death after excitotoxic/oxidative stress, and is considered to represent the opening of a pore in the mitochondrial membrane. There is evidence that the outer mitochondrial membrane protein porin is involved in the MPT and apoptosis. We present here a proof-of-principle study to address the hypothesis that anti-porin antibodies can prevent excitotoxic/ischemia-induced cell death. We generated anti-porin antibodies and show that the F(ab)(2) fragments penetrate living cells, reduce Ca(2+)-induced mitochondrial swelling as other MPT blockers do, and decrease neuronal death in dissociated and organotypic brain slice cultures exposed to excitotoxic and ischemic episodes. These observations present direct evidence that anti-porin antibody fragments prevent cell damage in brain tissue, that porin is a crucial protein involved in mitochondrial and cell dysfunction, and that it is conceivable that antibodies can be used as therapeutic agents. PMID- 12908926 TI - Treatment of cold injury-induced brain edema with a nonspecific matrix metalloproteinase inhibitor MMI270 in rats. AB - Blood-brain barrier (BBB) disruption is a critical event leading to vasogenic brain edema and secondary brain damage after cold injury-induced brain trauma. Matrix metalloproteinases (MMPs), a family of proteolytic enzymes which degrade the extracellular matrix, are implicated in BBB disruption in this model. The purpose of this study was to examine the effects of MMI270 (N-hydroxy-2(R)-[(4 methoxysulfony) (3-picolyl)-amino]-3-metylbutaneamide hydrochloride monohydrate), a synthetic nonspecific MMP inhibitor, on cold injury-induced brain edema in rats. Cold injury was induced by applying a copper probe cooled with liquid nitrogen on the parietal skull for 30 sec in 38 rats. Treatment with MMI270, a bolus injection at a dose of 30 mg/kg, was started immediately after the induction of cold injury and was continued for 24 h at a dose of 40 mg/kg/day using an intraperitoneal osmotic minipump (n = 7). In the untreated control group (n = 7), rats were administered a vehicle and implanted with a vehicle-containing osmotic pump. Two percent Evans Blue (EB) in saline (1 mL/kg) was administrated intravenously immediately after the cold injury in another group of rats, six of which were untreated and six of which were treated with MMI270 at the above dose. At 24 h after the cold injury, the brain water content and the BBB permeability to EB were determined. To assess the protective effect of MMI270 on secondary brain lesion after the cold injury, the MMI270-treated rats received a bolus injection at a dose of 30 mg/kg, followed by a continuous administration of MMI270 for 7 days at a dose of 40 mg/kg/day using an osmotic minipump (n = 6). In the untreated control group (n = 6), the rats were administered the vehicle and implanted with a vehicle-containing osmotic pump. At 7 days after cold injury, the secondary brain lesion was assessed using hematoxylin and eosin (H-E) staining. Compared with the untreated control group, treatment with MMI270 significantly reduced the brain water content in the ipsilateral core and intermediate areas (p < 0.05 and p < 0.01) and protected the BBB integrity to EB in the ipsilateral core area (p < 0.05) at 24 h after the cold injury. The secondary lesion was significantly smaller in the MMI270-treated animals compared with the untreated animals (p < 0.05) a 7 days after the cold injury. O kur results indicate that treatment with MMI270 in rats exhibits protection in acute brain edema formation and secondary brain damage by attenuating the BBB permeability after cold injury. PMID- 12908927 TI - Creatine diet supplement for spinal cord injury: influences on functional recovery and tissue sparing in rats. AB - Creatine-supplemented diet significantly attenuates cortical damage after traumatic brain injury in rodents. The protective mechanism likely involves maintenance of mitochondrial homeostasis. In the present study, we used two separate contusion spinal cord injury (SCI) instruments--the NYU device and the PSI Infinite Horizon (IH) impactor--to assess the efficacy of creatine supplemented diets on hind limb functional recovery and tissue sparing in adult rats. Rats were fed control versus 2% creatine-supplemented chow for 4-5 weeks prior to SCI (pre-fed), after which most resumed a control diet while some remained on a 2% creatine diet (pre & post-fed). Following long-term behavioral analysis (BBB), the amount of spared spinal cord tissue among the dietary regimen groups was assessed using stereology. Comparatively, both instruments caused similar amounts of gray matter damage while the NYU device rendered a greater loss of white matter, reflected in more severe hind limb functional deficits than with the IH impactor. Relative to the control fed groups injured with either instrument, none of the creatine fed animals showed improvements in hind limb function or white matter tissue sparing. Although creatine did not attenuate gray matter loss in the NYU cohort, it significantly spared gray matter in the IH cohort with pre-fed and pre & post-fed regimens. Such selective sparing of injured spinal cord gray matter with a dietary supplement yields a promising strategy to promote neuroprotection after SCI. The relationship between the efficacy of creatine and the magnitude of the insults is discussed. PMID- 12908928 TI - Macrophage recruitment follows the pattern of inducible nitric oxide synthase expression in a model for carpal tunnel syndrome. AB - Chronic nerve compression (CNC) induces a permeability change in neural vasculature. As recent evidence has shown that an alteration in reactive oxidative species (ROS) is related to neural degradation and regeneration, we evaluated whether inducible nitric oxide synthase (iNOS) plays a role in a rat model for CNC. Semi-quantitative analysis of iNOS mRNA and protein were performed with in situ hybridization and immunohistochemistry, respectively, at 3, 5, and 9 months post-operatively. At 3 months, iNOS mRNA was up-regulated in the perineurium of the proximal nerve with detectable changes in compressed and distal nerve segments. This expression continued to increase in the perineurium of 5-month proximal and compressed nerve segments with distal nerve demonstrating only a slight up-regulation of iNOS mRNA. At 9 months, iNOS mRNA expression was observed in both compressed and distal nerve. iNOS protein expression followed the same pattern of iNOS mRNA. As the perineurium is the blood-nerve barrier, the data suggests that these changes maybe mediated at the level of the perineurium. As macrophages release iNOS, we also evaluated whether macrophage recruitment followed the same pattern as iNOS expression. The results of ED-1 immunostaining for macrophages indicate that macrophages were localized to the outer one-third of cross sections during early time points. At later time points, macrophages were distributed diffusely throughout the nerve sections. Contrary to Wallerian degeneration, which elicits a relatively immediate signal for macrophage recruitment, CNC provides a slow, sustained stimulus for macrophage recruitment, which may be responsible for the up-regulation of iNOS gene expression. PMID- 12908929 TI - Median nerve trauma in a rat model of work-related musculoskeletal disorder. AB - Anatomical and physiological changes were evaluated in the median nerves of rats trained to perform repetitive reaching. Motor degradation was evident after 4 weeks. ED1-immunoreactive macrophages were seen in the transcarpal region of the median nerve of both forelimbs by 5-6 weeks. Fibrosis, characterized by increased immunoexpression of collagen type I by 8 weeks and connective tissue growth factor by 12 weeks, was evident. The conduction velocity (NCV) within the carpal tunnel showed a modest but significant decline after 9-12 weeks. The lowest NCV values were found in animals that refused to participate in the task for the full time available. Thus, both anatomical and physiological signs of progressive tissue damage were present in this model. These results, together with other recent findings indicate that work-related carpal tunnel syndrome develops through mechanisms that include injury, inflammation, fibrosis and subsequent nerve compression. PMID- 12908931 TI - pol gene sequence variation in Swedish HIV-2 patients failing antiretroviral therapy. AB - There is limited knowledge about how to treat and interpret results from genotypic resistance assays in HIV-2 infection. Here, genetic variation in HIV-2 pol gene was studied in 20 of 23 known HIV-2 cases in Sweden. Five patients with signs of virological treatment failure were longitudinally studied. Clinical, virological and immunological data were collected and the protease (PR) and first half of the reverse transcriptase (RT) was amplified and directly sequenced from plasma samples. Moderate to extensive genetic evolution was observed in four of the five patients who failed treatment. Some mutations occurred at positions known to confer resistance in HIV-1, but many occurred at other positions in PR and RT. All patients had been treated with zidovudine alone or in combination with other antiretroviral drugs, but none displayed a mutation at position 215, which is the primary zidovudine resistance site in HIV-1. Instead, a E219D mutation evolved in virus from two patients and a Q151M mutation evolved in two other patients. A M184V mutation indicative of lamivudine resistance was detected in three patients. The virus of one patient who had been treated with ritonavir, nelfinavir, and lopinavir successively acquired nine unusual mutations in the protease gene, most of which are not considered primary or secondary resistance mutations in HIV-1. Our data indicate that the evolutionary pathways that lead to antiretroviral resistance in HIV-2 and HIV-1 exhibit both similarities and differences. Genotypic HIV-2 resistance assays cannot be interpreted using algorithms developed for HIV-1, instead new algorithms specific for HIV-2 have to be developed. PMID- 12908932 TI - Broad spectrum of coreceptor usage and rapid disease progression in HIV-1 infected individuals from Central African Republic. AB - To study the progression of HIV-1 infection and coreceptor usages in Central African Republic, clinical data, plasma viral load, and coreceptor usage of sequential HIV-1 isolates were analyzed in a seroincident prospective cohort (PRIMOCA). Twenty-three HIV-1 infected individuals from the Central African Armed Forces were followed from 1995 to 2000. Viruses were isolated from 17 patients at various time points after seroconversion and their coreceptor usage was examined using GHOST cells expressing CD4 and one of the HIV-1 chemokine coreceptors CCR5, CXCR4, BOB/GPR15, and Bonzo/STRL33/CXCR6. Eleven patients died from AIDS. Eight of them died between 2 and 5 years after seroconversion, after a brief symptomatic stage. Patients who rapidly progressed to AIDS and death displayed the highest viral loads after seroconversion. All isolates obtained soon after seroconversion used CCR5, albeit, in some cases, CXCR4, BOB, or Bonzo were also used. Most isolates remained R5 (59 out of 61 isolates), although viruses using CXCR4 appeared in some cases of progression to AIDS. In several cases, a broad tropism was observed during the course of infection, with a frequent usage of BOB and Bonzo in addition to CCR5. Rapid progression to disease and short survival time among Central African HIV-1 patients appear more frequent than those reported in industrialized countries. Viral coreceptor used was mainly CCR5, but, interestingly, a large part of isolates also used BOB and Bonzo. However, there was no strict correlation between the clinical outcome and extended viral tropism. PMID- 12908933 TI - A new circulating recombinant form, CRF15_01B, reinforces the linkage between IDU and heterosexual epidemics in Thailand. AB - HIV-1 subtype B and CRF01_AE have been in circulation in Thailand and Southeast Asia for more than a decade. Initially separated by risk group, the two strains are increasingly intermixed, and two recombinant strains of essentially reciprocal structure have been recently reported. Here we identify additional CRF_01B recombinants and provide the evidence that HIV-1 strains now pass freely between the two high-risk populations. HIV isolates that showed discordance between CRF01_AE and subtype B in multi-region genotyping assays were selected for the study. They were drawn from 3 different cohorts in Thailand representing different risk behaviors and demographic characteristics: a drug user cohort in the north, a family planning clinic attendee cohort in the southeast, and a cohort study of the mucosal virology and immunology of HIV-1 infection in Thailand. The DNA from these isolates was PCR amplified to recover the full HIV-1 genome and subjected to sequencing and phylogenetic analysis. We establish that one particular CRF_01B recombinant, with the external envelope of subtype B and the rest of the genome from CRF01_AE, is circulating widely in Thailand. Termed CRF15_01B (also referred to as CRF15), the strain was primarily heterosexually transmitted, although injecting drug use (IDU) also played a role. In aggregate data from the studies, CRF15 constituted 1.7% of all HIV-1 infections (95% confidence interval 0.5-4.4%) and was dispersed widely in the country. The previously separate heterosexual and IDU epidemics have apparently been bridged by a new CRF. The entry of CRF15 into the mainstream of the epidemic signals new complexity in the long stable molecular picture in Thailand. These recombinants must be considered in ongoing or projected efficacy evaluations of HIV-1 vaccines and antiviral therapies. PMID- 12908934 TI - Heterogeneity of HIV-1 Rev response element. AB - We compiled all the RRE sequences of HIV-1 in the HIV Sequence Database and analyzed for base variation frequency at each nucleotide position. Positions with high frequency of base alteration scattered throughout the region, but primary sequences of almost all bases in stem IIA, Rev-binding bubble, and most of the stem region of stem-loop III were highly conserved. Comparing to HXB2 secondary structure, basepair-disrupting mutations did not distribute evenly in every region of the RRE. Stem I, stem IIB outside the Rev-binding site, stem IIC, and proximal parts of stem IV and V were more variable, while stem IIA, stem III, and distal parts of stem IV and V were highly conserved. These data indicated that RREs are structurally heterogeneous. The uneven distribution of variation in both primary sequence and the stem structure put forward highly conserved sites that might be more crucial to the function of RRE than the less conserved parts. PMID- 12908935 TI - Molecular epidemiology of dual HIV-1/HIV-2 seropositive adults from Senegal, West Africa. AB - Dual infection with HIV-1 and HIV-2 can occur in locales where these viruses co circulate, most commonly in West Africa. Although dual seropositivity is common in this region, the true rate of dual infection remains unclear. In addition, whether unique HIV-1 subtypes are circulating in dually infected individuals is unknown. A cohort of 47 HIV-1 and HIV-2 dually seropositive individuals from Senegal, West Africa was screened for the presence of HIV-1 and HIV-2 gag and env PBMC viral DNA sequences using PCR. Of the 47 dual HIV-1/HIV-2 seropositive individuals tested, 19 (40.4%) had infection with both HIV-1 and HIV-2 confirmed by genetic sequence analysis, whereas only HIV-1 or HIV-2 was confirmed in 17 (36.2%) or 9 (19.1%), respectively. The majority of HIV-1 subtypes found were CRF 02 and A, although subtypes D, C, G, J and B were also found, reflecting the subtypes known to be circulating in Senegal. There was no significant difference in HIV-1 subtype distribution between individuals with confirmed dual infection and patients in this study with dual seropositivity but lacking HIV-2, or with HIV-1 infected patients within the general population in Senegal, although the study was underpowered to detect anything but large differences. The prevalence of HIV-1/HIV-2 dual infection appears to be significantly less than that of dually seropositive individuals and this likely reflects cross-reactive serology. The common HIV-1 subtypes prevalent in West Africa (CRF-02 and subtype A) have a similar distribution to those found in our cohort of dually infected and dually seropositive subjects. PMID- 12908936 TI - Spectrum of manifestations of Mycobacterium tuberculosis infection in primates infected with SIV. AB - To characterize the manifestations of coinfection with M. tuberculosis and SIV infection, we studied 12 SIV-infected rhesus monkeys, six of which were infected intrabronchially with a low dose of Mycobacterium tuberculosis H37Rv. In the six coinfected animals, M. tuberculosis antigen-stimulated lung and blood cells produced high concentrations of IFN-gamma but not IL-4 8-16 weeks after infection. Of the three coinfected animals with high levels of plasma viremia, two developed disseminated tuberculosis and the other died of bacterial peritonitis. Of three coinfected animals with moderate levels of plasma viremia, two had no clinical or radiographic evidence of tuberculosis or progressive SIV infection for 6 months after infection. At neuropsy, pulmonary granulomata were observed and acid-fast organisms or M. tuberculosis were present. These clinical, immunologic and pathologic findings are consistent with those in humans with latent tuberculosis infection (LTBI), and suggest that a model of LTBI in SIV infected primates can be developed. Such a model will permit delineation of the immunologic and microbial factors that characterize LTBI in HIV-infected persons. PMID- 12908937 TI - Expression and functional activity of isotype and subclass switched human monoclonal antibody reactive with the base of the V3 loop of HIV-1 gp120. AB - Immunoglobulins undergo isotype switching in response to antigenic stimulation. The C(H) domains, in particular the hinge region, impose structural constraints on the interaction of antibody with antigen, especially multivalent antigens such as HIV. We previously showed that switching the IgG1 anti-HIV human monoclonal antibody (HMAb) F105 to an IgG3 resulted in significantly enhanced neutralization of HIV. To further investigate the influence of isotype, including the functional activity of HMAbs switched to IgA, which may be important in mucosal defenses, isotype switched antibodies have been generated for the anti-V3 loop base IgG2 HMAb F425B4e8. Reactivity of the IgG1 antibody was greater than the parental IgG2 antibody for SF2 infected cells but less for primary isolate virions. In contrast, there was less reactivity of the IgG3 with either infected cells or virions. IgA reacted significantly more with infected cells and virions as compared to the IgG subclasses. In contrast to previous studies whereby IgG3 enhanced neutralization, comparable neutralization of primary isolate virus was observed for IgG subclasses (IgG1, IgG2, IgG3) and IgA. This may reflect differences in the exposure of epitopes recognized by the HMAb with antibody flexibility being important to neutralization by antibodies reactive with obscured epitopes (e.g., CD4 binding site). Further analysis of the in vitro activity of isotype or subclass switched antibodies, IgA in particular, alone and in combination with other HMAbs, will provide important information on the role of IgG subclass and IgA antibodies on protective immunity to HIV. PMID- 12908938 TI - Comparison and evaluation of DRI methamphetamine, DRI ecstasy, Abuscreen ONLINE amphetamine, and a modified Abuscreen ONLINE amphetamine screening immunoassays for the detection of amphetamine (AMP), methamphetamine (MTH), 3,4 methylenedioxyamphetamine (MDA), and 3,4-methylenedioxymethamphetamine (MDMA) in human urine. AB - The performances of four immunoassays (DRI amphetamines, DRI ecstasy, Abuscreen ONLINE amphetamines, and a modified Abuscreen ONLINE amphetamines) were evaluated for control failure rates, sensitivity, and specificity for amphetamine (AMP), methamphetamine (MTH), 3,4-methylenedioxyamphetamine (MDA), and 3,4 methylenedioxymethamphetamine (MDMA). The two DRI reagents and the ONLINE reagents were run according to manufacturer specifications using a Roche Hitachi Modular DDP system. The modified ONLINE reagent was calibrated with MDMA and had 16mM sodium periodate added to the R2 reagent. These assays were run on approximately 27,500 human urine samples and 7000 control urine samples prepared at 350 and 674 ng/mL over the course of 8 days. All assays were calibrated using a single point, qualitative cutoff standard with the manufacturer-recommended compound at the Department of Defense cutoff (500 ng/mL). Gas chromatography-mass spectrometry (GC-MS) confirmation was conducted on screened-positive samples. Control performance for the manufacturer recommended assays was excellent, with a maximum qualitative control failure rate of 2.03%. The modified ONLINE reagent demonstrated poor control performance with a maximum failure rate of 38.3% and showed no improved MDMA sensitivity when compared with the ONLINE reagent; the confirmation rate (20%) was improved when compared with the production ONLINE reagent (8%). The DRI ecstasy reagent provided improved sensitivity for MDMA as compared with the ONLINE reagent, with approximately 23% more samples screening and confirming positive for MDMA and a confirmation rate of approximately 90%. The DRI methamphetamine reagent had a low confirmation rate (6% or less) and produced numerous positives for samples with only ephedrine or pseudoephedrine present. PMID- 12908939 TI - Quantitation of cocaine and its major metabolites in human saliva using gas chromatography-positive chemical ionization-mass spectrometry (GC-PCI-MS). AB - An analytical method for the simultaneous determination of cocaine (COC) and its major metabolites, ecgonine methyl ester (EME) and benzoylecgonine (BEG), in saliva was developed. The method involves liquid-liquid extraction in Toxitubes A, derivatization with 99:1 (v/v) N,O-bis-trimethylsilyltrifluoroacetamide (BSTFA)/trimethylchlorosilane (TMCS) and gas chromatography-mass spectrometry (GC MS) determination. The detector response thus obtained is linear over the range 25-1000 ng/mL, with a reproducibility better than 93% and a recovery close to 100% for the three analytes. The limits of detection achieved were 0.9 ng/mL for EME, 2.2 ng/mL for cocaine, and 0.2 ng/mL for BEG, and the limits of quantitation were 3.0 ng/mL for EME, 7.4 ng/mL for cocaine, and 0.8 ng/mL for BEG. The proposed method was applied to 48 saliva samples from cocaine users, 96% of which were positive for the drug and/or its metabolites. Saliva is thus a suitable biological fluid for determining cocaine, EME, and BEG by GC-PCI-MS. PMID- 12908940 TI - Direct and rapid determination of baclofen (Lioresal) and carisoprodol (Soma) in bovine serum by liquid chromatography-mass spectrometry. AB - Baclofen (Lioresal), a lipophilic analogue of c-aminobutyric acid (GABA), and carisoprodol (Soma), a central nervous system depressant with an unknown mechanism of pharmacologic action, are categorized as muscle relaxants. Baclofen is used clinically in the management of spasticity and its sequelae secondary to severe chronic disorders such as multiple sclerosis and other types of spinal cord lesions. Carisoprodol is used for discomfort associated with acute and painful musculoskeletal conditions. Intoxication from these drugs occurs in both humans and animals necessitating a need for their detection in plasma/serum, tissue, and gastrointestinal contents samples. A sensitive and specific analytical method for detection and quantitation of these compounds using liquid chromatography with positive atmospheric pressure chemical ionization-mass spectrometry was developed. A rapid extraction procedure for both analytes from fortified bovine sera is described. Chromatographic separation was carried out on a C(18) reverse-phase column with a gradient elution of acetonitrile and 0.25% acetic acid. The effluent was directed to the mass spectrometer with fragmentation information for baclofen and carisoprodol obtained in a scan monitoring mode. Linear standard curves for baclofen and carisoprodol were constructed based on at least two corresponding extracted ions over a concentration range of 0.1-50 micro g/mL. The analysis of fortified sera samples demonstrates good accuracy and precision for the method with a limit of detection of 0.5 micro g/mL for carisoprodol (n = 3) and 1 micro g/mL for baclofen (n = 4) and a limit of quantitation of 2 micro g/mL for both compounds. Recoveries at the limit of quantitation were between 75 and 95% for both analytes, with a 4.8-9.3% range in standard deviation. PMID- 12908941 TI - Quantitative detection of nine phthalate metabolites in human serum using reversed-phase high-performance liquid chromatography-electrospray ionization tandem mass spectrometry. AB - We developed a highly sensitive method for the quantitative detection of nine phthalate ester metabolites in human serum. This method requires denaturation of the serum enzymes immediately after blood collection to avoid the hydrolysis of the contaminant diester parent compounds introduced during blood collection and storage. Before analysis, the samples were subjected to an enzymatic deconjugation to hydrolyze the glucuronidated phthalate monoesters and a solid phase extraction to isolate the monoesters from other serum components. The extracts were analyzed using reversed-phase high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry. The limits of detection of all nine phthalate monoesters in serum were in the low nanogram-per milliliter range (0.6-1.3 ng/mL). Stable isotope-labeled internal standards for all analytes were used to improve precision and for recovery corrections. This highly selective method permits the analysis of phthalate monoesters without interferences resulting from the hydrolysis of the ubiquitous contaminant phthalate diesters by serum enzymes. In addition, it allows the direct measurement of the active phthalate monoester metabolites reportedly responsible for the reproductive and developmental toxicity of certain phthalates. PMID- 12908942 TI - Distribution of serum paraoxon hydrolyzing activity in a large Spanish population using a routine automized method in clinical laboratory. AB - This work was performed to adapt the manual laboratory method of measuring serum paraoxonase activity using a routine automatized method in the clinical laboratory and to study the distribution of paraoxonase activity in a large population from Alcoy, a region of Spain. The serum samples for the study were obtained from extractions of blood from 2891 individuals, distributed by sex and age groups, in a routine check in a primary care facility of Alcoy. Paraoxonase activity was assayed by measuring the release of p-nitrophenol according to a previously published method adapted to an automatized analyzer. The mean paraoxonase activity recorder was 70.2 +/- 16.5 IU/L. Paraoxonase activity in children (both males and females) was significantly lower (p < 0.0005) than in older individuals. Paraoxonase activity detected in males and females older than 56 was slightly lower than that detected in younger individuals, although in this case the difference was not statistically significant. The paraoxonase activity shows higher mean values in females than in males (p < 0.0005). Human paraoxonase activity shows a unimodal distribution pattern in the studied population, which is in contrast with other studies showing bimodal distribution. PMID- 12908943 TI - Usefulness of sweat testing for the detection of MDMA after a single-dose administration. AB - Nine healthy male subjects and recreational users of 3,4 methylenedioxymethamphetamine (MDMA) participated in a study aimed to assess the usefulness of sweat testing for the detection of MDMA after a single 100-mg dose. Sweat was collected for up to 24 h with the PharmChek sweat patches from which drugs were eluted and then analyzed by immunoassay and gas chromatography-mass spectrometry using deuterated internal standards. The usefulness of a rapid onsite test, the Drugwipe immunochemical strip test, was also assessed. In the sweat patches, MDMA was detected as early as 1.5 h after consumption and peaked at 24 h. Intersubject variability was large; peak MDMA concentrations for the same dose varied in magnitude 30-fold. MDMA concentrations ranged between 3.2 and 1326.1 ng/patch. Only traces of the minor metabolite 3,4 methylenedioxyamphetamine were detected. In all subjects, the onsite test with the Drugwipe was positive at 1.5 h (peak time of MDMA plasma concentration). However, few false-negative results (18%) appeared in the first 6 h after administration. Both sweat patch testing and the onsite sweat strip test may find useful application for noninvasive monitoring of MDMA abuse in sweat. PMID- 12908944 TI - Simultaneous plasma lamotrigine analysis with carbamazepine, carbamazepine 10,11 epoxide, primidone, phenytoin, phenobarbital, and PEMA by micellar electrokinetic capillary chromatography (MECC). AB - The determination of lamotrigine (LTG) simultaneously with carbamazepine (CBZ), carbamazepine 10,11 epoxide (CBZ-E), primidone (PRM), phenytoin (PHT), phenobarbital (PB), and 2-phenyl-2-ethyl-malonamide (PEMA) in human plasma was developed using micellar electrokinetic capillary chromatography (MECC) with a diode-array detector. The reproducibility of both separation and quantitation with MECC analysis were appropriate for the intra- and interassay coefficients. The evaluated drugs concentration intervals of LTG, 0.5-10.0 micro g/mL; CBZ, 1.0 16.0 micro g/mL; PEMA, 1.0-20.0 micro g/mL; PB, 1.0-60.0 micro g/mL; PRM, 1.0 20.0 micro g/mL; PHT, 0.7-40.0 micro g/mL; and CBZ-E, 1.0-14.0 micro g/mL were linear with correlation coefficients higher than 0.987 and coefficients of the variation of the points of the calibration curve lower than 10%. The limit of quantitation of the investigated drugs in plasma varied from 0.5 to 1.0 micro g/mL, depending upon the drug. The MECC technique was sensitive enough to work with microsamples into the subtherapeutic, therapeutic, and toxic concentrations, as well as showed to be simple and efficient when applied to monitoring therapeutic drugs in patients treated with a combination of lamotrigine and other antiepileptic drugs such as hepatic enzyme-inducing agents. PMID- 12908945 TI - Direct analysis of plasticizers in aqueous samples by atmospheric pressure chemical ionization-tandem mass spectrometry (APCI-MS-MS). PMID- 12908946 TI - Foxy, a designer tryptamine hallucinogen. AB - Foxy is slang for 5-methoxy-N,N-diisopropyltryptamine. It has hallucinogenic properties, similar to other tryptamine compounds, and is mildly euphoric. This case report describes a 21-year-old Caucasian man who ingested a pill called Foxy containing an unknown amount of drug. He was observed in hospital for 2 h, during which time he had mild hallucinations and could not move his limbs. A urine sample was collected approximately 4 h after drug ingestion. The patient was then discharged with no follow up assessment. The 5-methoxy-N,N-diisopropyltryptamine was identified in the urine by gas chromatography-mass spectrometry. Standards prepared from the pure material were used in the identification. Quantitative analysis using the same analytical technique resulted in a urinary concentration of 1.7 micro g/mL. Through oxidative deamination, the metabolite, 5-methoxy indole acetic acid, was formed. It was identified in the urine, and the concentration was determined to be 1.3 micro g/mL using gas chromatography-mass spectrometry. Two other compounds were discovered in the urine sample as a result of a routine drug screen. From their mass spectra, they were tentatively identified as 5-methoxy-N-isopropyltryptamine and 5-methoxy-N,N diisopropyltryptamine-N'-oxide. PMID- 12908947 TI - Fatality due to combined use of the designer drugs MDMA and PMA: a distribution study. AB - We present a fatal case involving the combined ingestion of amphetamine, 3,4 methylenedioxymethylamphetamine, 3,4-methylenedioxyamphetamine, and paramethoxyamphetamine. Various postmortem specimens (e.g., several blood samples, urine, and tissue samples) were analyzed to study the distribution of the compounds and their metabolites in the human body. Quantitation took place using liquid chromatography-sonic spray ionization-mass spectrometry after pretreatment with a liquid-liquid extraction. The medico-legal findings were compatible with a disseminated intravascular coagulation induced by hyperthermia caused by the simultaneous intake of the amphetamine analogues. PMID- 12908949 TI - Evaluation of a new reagent: anthraquinone-2-sulfonyl chloride for the determination of phenol in water by liquid chromatography using precolumn phase transfer catalyzed derivatization. AB - A new reagent, anthraquinone-2-sulfonyl chloride, is used for the derivatizaton of phenols. Several compounds with different polarities are selected to evaluate the new reagent and derivatives of these phenols that are prepared via a facile pathway. The optimal conditions for analytical derivatization and mechanism of the derivatization reaction are discussed. The derivatization procedure involves an ion-pair extraction of the deprotonated phenols with a tetrabutylammonium counter ion in the organic phase. At the interface of two phases, the derivatization reaction occurs quantitatively at room temperature within 3 min. The derivatives are stable and readily amenable to analysis by normal-phase (NP) and reversed-phase (RP) high-performance liquid chromatography (HPLC). Excellent linearity response was demonstrated over the concentration range of 0.2-200 micromol/L at 320 nm for NP-HPLC and at 256 nm for RP-HPLC. Combined with preconcentration using a Waters Sep-Pak Plus C(18) cartridge, detection limits of phenols for water-sample analysis are as low as 1 x 10(-9) mol/L (approximately 0.1 microg/mL). PMID- 12908948 TI - Fatal acute topiramate toxicity. AB - A 44-year-old Caucasian female was found dead in bed. Qualitative screening detected ethanol, phenobarbital, and methotrimeprazine. However, none were sufficient to attribute as the cause of death. Additionally, high concentrations of topiramate, an antiepilieptic agent, were found. Analysis of available biological fluids and tissues was carried out with the following results: blood (central) 170 mg/L, liver 140 mg/kg, stomach contents greater than 300 mg, and vitreous fluid 65 mg/L. The cause of death was ascribed to topiramate overdose. PMID- 12908950 TI - Multiresidue determination of acidic pesticides in water by HPLC-DAD with confirmation by GC-MS using conversion to the methyl ester with trimethylsilyldiazomethane. AB - A multiresidue pesticide methodology is studied and results for acidics are reported here, with base/neutral to follow. This work studies a literature procedure as a possible general approach to many pesticides and potentially other analytes that are considered to be liquid chromatographic (LC) candidates, rather than gas chromatographic (GC) ones. The analysis of the sewage effluent of a major southwestern U.S. city serves as an example of the application of the methodology to a real sample. Recovery studies were also conducted to validate the proposed extraction step. A gradient elution program was followed for the high-performance LC (HPLC) leading to a general approach for acidics. Confirmation of identity was by electron ionization-GC-mass spectrometry (MS) after conversion of the acids to the methyl ester (or other appropriate methylation) by means of trimethylsilyldiazomethane. The 3,4 dichlorophenoxyacetic acid is used as an internal standard to monitor the reaction, and polychlorobiphenyl #19 is used for the quantitation internal standard. Although others have reported similar analyses of acids, conversion to the methyl ester is by means of diazomethane itself, rather than by the more convenient and safe trimethylsilyldiazomethane. Thus, the present paper supports the use of trimethylsilyldiazomethane with all of these acids (trimethylsilyldiazomethane has been used in environmental work with some phenoxyacetic acid herbicides) and further supports the usefulness of this reagent as a potential replacement for diazomethane. The HPLC approach here could also serve as the separation basis for an LC-MS solution for confirmation of identity, as well as quantitation. PMID- 12908951 TI - Improvement of peak shape and separation performance of beta-blockers in conventional reversed-phase columns using solvent modifiers. AB - A comparative study of peak shape, elution behavior, and resolution of 16 beta blockers (acebutolol, alprenolol, atenolol, bisoprolol, carteolol, celiprolol, esmolol, labetalol, metoprolol, nadolol, oxprenolol, pindolol, practolol, propranolol, sotalol, and timolol) chromatographed with hybrid mobile phases of triethylamine (TEA)-acetonitrile and sodium dodecyl sulfate (SDS)-propanol is performed using conventional reversed-phase columns and isocratic elution. Both solvent modifiers (TEA and SDS) prevent the interaction of the basic drugs with the alkyl-bonded phase. However, the protection mechanisms of silanols on the packing are different. Whereas TEA associates with the silanol sites (blocking ion-exchange processes or repelling the solutes), the long hydrophobic chain of SDS is inserted in the bonded organic layer with the sulfate group protruding outside, which makes the stationary phase negatively charged. The effects of TEA, acetonitrile, SDS, and propanol on the elution strength, efficiency, peak asymmetry, and resolution are examined under an experimental design basis that is assisted by computer simulation to reach more general conclusions. The combination of improved peak shapes, larger selectivity, and a smaller range in retention among compounds of extreme polarity leads to the observation that a greater number of beta-blockers can be resolved with a hybrid micellar system. PMID- 12908952 TI - Densitometric thin-layer chromatographic determination of artemisinin and its lipophilic derivatives, artemether and arteether. AB - A thin-layer chromatograpy (TLC) method is developed to analyze artemisinin (AT) and its derivatives, artemether (AM) and arteether (AE), using a silica-gel plate with a mobile phase containing pure chloroform. After development, all products are visualized after dipping in a 4-methoxybenzaldehyde dipping reagent of 1% (v/v) in an acidic solution of sulphuric acid (98%, v/v) and acetic acid (96-98%, v/v) (respectively, 2% and 10%, v/v in alcohol-water, 60:30, v/v), presenting a purple color against a slightly colored background. This TLC system is quantitatively evaluated in terms of stability of the color, precision, accuracy, and calibration. Activation is performed at 110 degrees C. Stability of the color of both analytes is reached after 12 min. Precision, less than 5%, is obtained at two levels. Good linearity is obtained in the range of 0.5-8 micro g for all analytes. Some applications show its utility in the quality control of capsules. The prederivatization technique, applying the described dipping reagent before development, reveals the presence of various reaction products, possibly isomers. These results prove that TLC can be a cheap and easy alternative for the analysis of AT and its lipophilic derivatives, AM and AE, as pure powder and in pharmaceutical-dosage forms. PMID- 12908953 TI - Measurement of As, P, and S in the waste gases and water emitted from semiconductor processes by high-temperature hydrogen reduction gas chromatography. AB - A quick, sensitive, and accurate method, high-temperature hydrogen reduction gas chromatography (GC) (1,2), for measuring arsenic (As), phosphorus (P), and sulfur (S) in the waste gases and water emitted from semiconductor processes is proposed in this paper. A high-temperature hydrogen reduction system that changes As, P, S, and their compounds into hydrides by atomic hydrogen has been designed. It is convenient to detect these elements in solid, liquid, and gaseous samples by high temperature hydrogen reduction GC without pretreating samples. The lower detection limits of As, P, and S by this method are 0.01, 0.003, 0.02 mg/L, respectively, and the values of relative standard deviation are 6.2%, 8.6%, and 0.3%, respectively. Results determined by high-temperature hydrogen reduction GC are primarily accordant to those by conventional methods such as colorimetry and ion chromatography. The error statistics of this analysis method also show that high-temperature hydrogen reduction GC can be successfully used to determine trace As, P, and S in waste gases and wastewater emitted from semiconductor processes. PMID- 12908954 TI - A validated HPLC method for the assay of xanthone and 3-methoxyxanthone in PLGA nanocapsules. AB - This work relates the development and validation of a simple reversed-phase high performance liquid chromatographic (HPLC) method for the analysis of xanthone (XAN) and 3-methoxyxanthone (3-MeOXAN) in poly(D,L-lactide-co-glycolide) (PLGA) nanocapsule formulations. This method does not require any complex sample extraction procedure. Chromatographic separation is made with a reversed-phase C(18) column, using methanol-water (90:10, v/v) as a mobile phase at a flow rate of 1 mL/min. Identification is made by UV detection at 237 nm. The isocratic system operates at ambient temperature and requires 7 min of chromatographic time. The developed method is statistically validated according to United States Pharmacopoeia 25 and International Conference on Harmonization guidelines for its specificity, linearity, accuracy, and precision. The assay method proposed in this study is specific for XAN and 3-MeOXAN in the presence of nanocapsule excipients. Diode-array analyses confirm the homogeneity of XAN and 3-MeOXAN peaks in stressed conditions. Standard curves are linear (r > 0.999) over the concentration range of 0.4-2.5 and 1.0-5.8 micro g/mL for XAN and 3-MeOXAN, respectively. Recovery from nanocapsules ranges from 99.6% to 102.8% for XAN and 98.8% to 102.4% for 3-MeOXAN. Repeatability (intra-assay precision) is acceptable with relative standard deviation values of 1.2% for XAN and 0.3% for 3-MeOXAN. PMID- 12908955 TI - A clean and rapid liquid chromatographic technique for sulfamethazine monitoring in pork tissues without using organic solvents. AB - A rapid method for the isolation and high-performance liquid chromatographic (HPLC) determination of sulfamethazine (SMZ) in pork tissues (kidney, liver, and muscle) without using organic solvents is developed. The isolation is performed by homogenization with an acid solution using an ultrasonic-homogenizer, followed by centrifugation. The HPLC analyses are performed using a reversed-phase C(4) column (150- x 4.6-mm i.d.), a mobile phase of 0.02 mol/L citric acid solution, and a photodiode array detector. The resulting HPLC chromatograms are free from interferences for determination and identification. The proposed technique is shown to be linear (r > 0.99) over the concentration range 0.1-2.0 microg/g for all pork tissues. Average recoveries of SMZ (spiked 0.1-2.0 microg/g) range from 87.6% to 90.2%, with inter- and intra-assay variabilities of less than 4%. The total time required for the analysis of one sample and limit of quantitation is less than 20 min and 0.09 microg/g, respectively. PMID- 12908956 TI - Quantitation of tolmetin by high-performance liquid chromatography and method validation. AB - A high-performance liquid chromatographic (HPLC) assay method for assessing the degradation of tolmetin (TLM) is developed and validated under acidic, basic, and photoirradiated conditions. The HPLC method includes an Inertsil 5 ODS-3V column (250- x 4.6-mm i.d.), guard column of Inertsil 7 ODS-3V (50- x 4.6-mm i.d.), mobile phase of CH(3)OH-1% HOAc (64:36, v/v), and UV detection at 254 nm. The developed method satisfies the system suitability criteria, peak integrity, and resolution for the parent drug and its degradants. The established assay method exhibits good selectivity and specificity suitable for stability measurements. From the intra- and interday tests of six replicates, the coefficients of variation are between 0.20% and 1.77% for the former, and 0.12% and 3.40% for the latter. Recoveries are found to be 98.7-101.7%. TLM is determined to be more reactive when exposed to light and acidic conditions, yet TLM is stable in a basic medium. A kinetic study of the photodegradation of TLM shows that it follows an apparent first-order reaction in three alcoholic solvents. PMID- 12908957 TI - A comparative study of the ability of different techniques to extract menthol from Mentha piperita. AB - Supercritical fluid extraction, direct thermal desorption, hydroalcoholic extraction, and atomization are used to extract menthol from leaf plants of Mentha piperita. The investigated methods are comparatively evaluated on the basis of their reliability to determine the enantiomeric distribution of menthol. The enantioselectivity required for the gas chromatographic analysis is achieved using Octakis (2,6-di-O-n-pentyl-3-O-butyryl)-gamma-cyclodextrin as the stationary phase. From the obtained results, it is established that there is a significant effect of the combination of pressure and temperature to achieve the effective isolation and fractionation of the less and most volatile compounds using supercritical fluids. PMID- 12908960 TI - Arthropod containment guidelines. A project of the American Committee of Medical Entomology and American Society of Tropical Medicine and Hygiene. PMID- 12908965 TI - Promoter interference mediated by the U3 region in early-generation HIV-1-derived lentivirus vectors can influence detection of transgene expression in a cell-type and species-specific manner. AB - In a previous study using an early-generation VSV-G-pseudotyped lentivirus vector encoding enhanced green fluorescent protein (EGFP) under the transcriptional control of a human cytomegalovirus (CMV) immediate-early promoter, we examined transduction efficiency in dissociated dorsal root ganglia (DRG) cultures. In cultures of murine origin, transgene expression was observed solely in the sensory neurons with the stromal cell population failing to show evidence of transduction. In contrast, efficient and sustained transduction of both sensory neurons and the stromal cell population was observed in cultures of human origin. Given the widespread use of murine models in preclinical gene therapy studies, in the current study we investigated the basis of this apparent neuron specificity of lentivirus-mediated transduction in murine DRG cultures. The interspecies differences persisted at high multiplicities of infection, and irrespective of whether lentiviral vector stocks were packaged in the presence or absence of human immunodeficiency virus type 1 (HIV-1) accessory proteins. Cell-type specificity of CMV promoter expression, tropism of the VSV-G envelope, and blocks to molecular transduction were also precluded as possible mechanisms, thereby implicating transcriptional repression of the internal heterologous promoter. This promoter interference effect was found to be mediated by cis-acting sequences upstream of the core promoter elements located in the U3 region of the proviral long terminal repeats (LTRs). Deletion of this region, as in late generation self-inactivating (SIN) lentivirus vectors, relieves this effect. This provides a basis for reevaluating data produced using early-generation U3-bearing lentivirus vectors and for reconciling these with results obtained using more contemporary SIN lentivirus vectors carrying a U3 deletion. PMID- 12908966 TI - Size-exclusion chromatography purification of high-titer vesicular stomatitis virus G glycoprotein-pseudotyped retrovectors for cell and gene therapy applications. AB - Vesicular stomatitis virus G glycoprotein (VSV-G)-pseudotyped replication defective retroviral particles are pantropic and amenable to concentration to high titer by ultracentrifugation. These features have allowed development of effective retroviral transduction protocols for stem cells in vitro as well as for tissue engineering in vivo. However, retroparticle ultracentrifugation protocols will also copellet cellular and subcellular debris released from retroviral producer cell lines during vector manufacture. We have analyzed concentrated vector preparations by chromatography and have found that a significant amount of genomic DNA released from producer cells coconcentrates with retroviral particles. In an effort to generate high-purity retroparticle preparations, devoid of subcellular contaminants and contaminating genomic DNA, we have developed a process using size-exclusion chromatography combined with host cell nucleic acid digestion and concentration by ultrafiltration. The procedure allowed for a final recovery of 19 +/- 0.4% infectious viral particles from unfractionated starting material, with an average retroparticle concentration of 7.7 x 10(7) +/- 1.5 x 10(6)/ml. The intact virus is of high purity, >90% as determined by anion-exchange high-performance liquid chromatography. Retroparticle structure appeared intact as determined by negative stain electron microscopy and purified virus was functional and allowed for efficient transduction of primary human bone marrow stromal cells in vitro. In conclusion, we have developed a VSV-G retrovector purification process that can be applied to large-scale retroviral production ideal for cell and gene therapy applications. PMID- 12908967 TI - Retroviral vectors for high-level transgene expression in T lymphocytes. AB - Efficient expression of genes transferred by retroviral vectors is a prerequisite for gene therapy, especially when the biological effect depends on the amount of transgene product. High-level gene expression is desirable for several gene therapy approaches involving T lymphocytes. We evaluated standard retroviral vectors with cis-regulatory control elements of the Moloney murine leukemia virus (Mo-MLV) with or without the human T cell-specific CD2 enhancer. For comparison, vectors containing the long terminal repeat (LTR) of myeloproliferative sarcoma virus (MPSV) and an improved 5' untranslated region were used (MP71 vectors), with or without the woodchuck hepatitis virus posttranscriptional regulatory element (PRE). All vectors expressed the enhanced green fluorescent protein (GFP) to measure transgene expression. In mouse T cells MP71 vectors with and without the PRE yielded an up to 10-fold higher expression level compared with the Mo-MLV based vectors currently used for gene transfer into T lymphocytes. A high multiplicity of infection (MOI) of standard Mo-MLV vectors could not reach expression levels obtained with a low MOI of MP71 vector. Ex vivo-transduced mouse T lymphocytes maintained the vector-dependent differences in level of transgene expression in Rag-1-deficient mice when adoptively transferred. In four human T cell lines and human primary T lymphocytes MP71 vectors yielded an up to 75-fold higher GFP expression level in comparison with the standard Mo-MLV vector. In contrast to mouse T cells, the integration of the PRE into MP71 vectors induced in human T cells a further significant increase in transgene expression level. Southern blot analysis of CEM T cells revealed that the superior performance of MP71 vectors was not due to a higher rate of viral integration. In summary, MP71 vectors are useful tools for stable, high-level gene expression in T lymphocytes, for example, in the expression of T cell receptor genes. PMID- 12908968 TI - Extended amplification in vitro and replicative senescence: key factors implicated in the success of human myoblast transplantation. AB - The limited success of human myoblast transplantation has been related to immune rejection, poor survival, and limited spread of injected myoblasts after transplantation. An important issue that has received little attention, but is nevertheless of fundamental importance in myoblast transplantation protocols, is the proliferative capacity of human satellite cells. Previous studies from our laboratory have demonstrated that the maximum number of divisions that a population of satellite cells can make decreases with age during the first two decades of life then stabilizes in adulthood. These observations indicate that when satellite cells are used as vectors in myoblast transplantation protocols it is important to consider donor age and the number of divisions that the cells have made prior to transplantation as limiting factors in obtaining an optimal number of donor derived muscle fibers. In this study, myoblasts derived from donors of different ages (newborn, 17 years old, and 71 years old) were isolated and amplified in culture. Their potential to participate in in vivo muscle regeneration in RAG2(-/-)/gamma(c)/C5 triple immunodeficient hosts after implantation was evaluated at 4 and 8 weeks postimplantation. Our results demonstrate that prolonged amplification in culture and the approach to replicative senescence are both important factors that may condition the success of myoblast transplantation protocols. PMID- 12908969 TI - In vivo delivery of glial cell-derived neurotrophic factor across the blood-brain barrier by gene transfer into brain capillary endothelial cells. AB - The gene encoding mouse glial cell-derived neurotrophic factor (mGDNF gene) was transfected into brain capillary endothelial cells (BCECs) with the aim of delivering the gene product extensively into the brain parenchyma by making use of the secretory function of BCECs. First, we transfected mGDNF gene into cultured BCECs (MBEC4; mouse brain capillary endothelial cells) in vitro. The amount of mGDNF protein secreted from the transfected cells into the medium was 1500 to 3200 pg/mg of cell protein per day, being about sevenfold higher than that accumulated intracellularly. Furthermore, the basolateral-directed secretion of mGDNF protein from the transfected MBEC4 cells was fivefold higher than the apical-directed secretion. Next, the hemagglutination virus of Japan (HVJ) liposomes encapsulating mGDNF gene were administered to rats in vivo via the internal carotid artery. The transfected rats showed a marked increase in the brain level of GDNF as assessed by means of enzyme-linked immunosorbent assay (ELISA) and Western blotting on day 3 after the administration, and the level remained significantly elevated for at least 12 days. Furthermore, immunohistochemical staining revealed an increase in GDNF immunoreactivity throughout the transfected forebrain. These results indicate that the gene was successfully transferred in vivo from HVJ-liposomes into BCECs, where it was expressed, and the gene product was secreted into the brain. Then, using this delivery method, we evaluated the protective effect for dopamine neuron against a retrograde 6-hydroxydopamine (6-OHDA) lesion, as assessed by behavioral and neurochemical indices. PMID- 12908970 TI - In vivo targeting of tumor endothelial cells by systemic delivery of lentiviral vectors. AB - Tumor angiogenesis is a rate-limiting factor for tumor growth, and the endothelial cells of tumor vessels display specific features that can be exploited for the selective delivery of cancer therapeutics. To specifically target exogenous genes to angiogenic tumor vessels, we generated a panel of vesicular stomatitis virus-pseudotyped lentiviral vectors (LVs) engineered for endothelial cell (EC)-specific expression. We cloned a wide repertoire of transcription regulatory sequences from genes preferentially expressed in ECs (Tie1, Tie2, Flk-1, VE-Cad, and ICAM-2) into self-inactivating LVs to drive expression of the marker gene encoding green fluorescent protein (GFP) or of the conditionally toxic gene encoding nitroreductase, and compared them with the ubiquitously expressing phosphoglycerate kinase (PGK) and cytomegalovirus (CMV) promoters. We evaluated the efficiency and specificity of vector expression in vitro in a panel of human primary cultures, including ECs, fibroblasts, neurons, lymphocytes, and hematopoietic progenitors, and in tumor cell lines. We found that vectors containing promoter and enhancer sequences from the Tie2 gene achieved remarkable specificity of expression in ECs in vitro and in vivo. On intravenous delivery into tumor-bearing mice, the Tie2 vector targeted expression to the ECs of tumor vessels. In contrast, LVs carrying the PGK or CMV promoter gave widespread GFP marking in ECs and non-ECs of tumors and other organs. The previously reported upregulation of the Tie2 gene in ECs activated for angiogenesis may explain the remarkable selectivity of expression of the Tie2 vector in ECs of tumor vessels. The new vector provides the means for selective delivery of gene therapy to tumor sites in vivo. PMID- 12908971 TI - Efficient lentiviral vectors for short hairpin RNA delivery into human cells. AB - RNA interference is an evolutionarily conserved process of gene silencing that in plants serves as a natural defense mechanism against exogenous viral agents. RNA interference is becoming an important tool for the study of biological processes through reverse genetics and has potential for therapeutic applications in humans; however, effective delivery is still a major issue. Small interfering RNA (siRNA) and short hairpin RNA (shRNA) have been introduced into cells by transfection of chemically synthesized and RNA expression via plasmid cassettes utilizing RNA polymerase III transcription. The employment of siRNA/shRNA for gene knockout requires an efficient stable transfection or transduction process. Here, we report the successful construction of lentiviral vectors to express shRNA stably in human cells. We demonstrate that lentiviral vectors expressing siRNA directed to the reporter gene luciferase, when stably transduced into human cells without drug selection, are capable of protecting the cells from infection by a lentiviral vector encoding humanized firefly luciferase as a reporter gene. We observed 16- to 43-fold reduction of gene expression in infected cells transduced with shRNA vectors relative to cells transduced with control vectors. This model system demonstrates the utility of lentiviral vectors to stably express shRNA as both a cellular gene knockout tool and as a means to inhibit exogenous infectious agents such as viruses in human cells. PMID- 12908973 TI - Reproduction and longevity among the British peerage: the effect of frailty and health selection. AB - Whether a cost of reproduction exists among humans is still questionable. A major study of aristocratic British families finds a significant positive correlation between parity and late-life mortality, which indicates a trade-off between reproduction and longevity. This result is supported by four other studies, while earlier studies have not found a relationship or came to the opposite conclusion. We show that in natural fertility populations the relationship between fertility and late-life mortality cannot be studied correctly without considering the effects of differences in health and of mortality selection during childbearing ages because these two effects lead to a dampening of the true relationship. If these effects are controlled in Hollingsworth's genealogy of the British peerage a significant trade-off between reproduction and longevity exists for females but not for males. PMID- 12908974 TI - Explaining and predicting patterns in stochastic population systems. AB - Lattice effects in ecological time-series are patterns that arise because of the inherent discreteness of animal numbers. In this paper, we suggest a systematic approach for predicting lattice effects. We also show that an explanation of all the patterns in a population time-series may require more than one deterministic model, especially when the dynamics are complex. PMID- 12908975 TI - Is hippocampal volume affected by specialization for food hoarding in birds? AB - The hypothesis that spatial-memory specialization affects the size of the hippocampus has become widely accepted among scientists. The hypothesis comes from studies on birds primarily in two families, the Paridae (tits, titmice and chickadees) and the Corvidae (crows, nutcrackers, jays, etc.). Many species in these families store food and rely on spatial memory to relocate the cached items. The hippocampus is a brain structure that is thought to be important for memory. Several studies report that hoarding species in these families possess larger hippocampi than non-hoarding relatives, and that species classified as large-scale hoarders have larger hippocampi than less specialized hoarders. We have investigated the largest dataset on hippocampus size and food-hoarding behaviour in these families so far but did not find a significant correlation between food-hoarding specialization and hippocampal volume. The occurrence of such an effect in earlier studies may depend on differences in the estimation of hippocampal volumes or difficulties in categorizing the degree of specialization for hoarding or both. To control for discrepancies in measurement methods we made our own estimates of hippocampal volumes in 16 individuals of four species that have been included in previous studies. Our estimates agreed closely with previous ones, suggesting that measurement methods are sufficiently consistent. Instead, the main reasons that previous studies have found an effect where we did not are difficulties in assessing the degree of hoarding specialization and the fact that smaller subsets of species were compared than in our study. Our results show that a correlation between food-hoarding specialization and hippocampal volume cannot be claimed on the basis of present data in these families. PMID- 12908976 TI - Evidence for gene flow via seed dispersal from crop to wild relatives in Beta vulgaris (Chenopodiaceae): consequences for the release of genetically modified crop species with weedy lineages. AB - Gene flow and introgression from cultivated to wild plant populations have important evolutionary and ecological consequences and require detailed investigations for risk assessments of transgene escape into natural ecosystems. Sugar beets (Beta vulgaris ssp. vulgaris) are of particular concern because: (i) they are cross-compatible with their wild relatives (the sea beet, B. vulgaris ssp. maritima); (ii) crop-to-wild gene flow is likely to occur via weedy lineages resulting from hybridization events and locally infesting fields. Using a chloroplastic marker and a set of nuclear microsatellite loci, the occurrence of crop-to-wild gene flow was investigated in the French sugar beet production area within a 'contact-zone' in between coastal wild populations and sugar beet fields. The results did not reveal large pollen dispersal from weed to wild beets. However, several pieces of evidence clearly show an escape of weedy lineages from fields via seed flow. Since most studies involving the assessment of transgene escape from crops to wild outcrossing relatives generally focused only on pollen dispersal, this last result was unexpected: it points out the key role of a long-lived seed bank and highlights support for transgene escape via man-mediated long-distance dispersal events. PMID- 12908977 TI - Transients and attractors in epidemics. AB - Historical records of childhood disease incidence reveal complex dynamics. For measles, a simple model has indicated that epidemic patterns represent attractors of a nonlinear dynamic system and that transitions between different attractors are driven by slow changes in birth rates and vaccination levels. The same analysis can explain the main features of chickenpox dynamics, but fails for rubella and whooping cough. We show that an additional (perturbative) analysis of the model, together with knowledge of the population size in question, can account for all the observed incidence patterns by predicting how stochastically sustained transient dynamics should be manifested in these systems. PMID- 12908978 TI - Assessment of the risk posed by bovine spongiform encephalopathy in cattle in Great Britain and the impact of potential changes to current control measures. AB - We extended an existing back-calculation model to analyse data on reported clinical cases of bovine spongiform encephalopathy (BSE), data from random testing of healthy animals slaughtered in abattoirs and testing data from animals reported as sick or dying on the farm. Extensive analysis of demographic data was also undertaken. We estimated past and current BSE infection prevalences in the cattle population and the degree of case under-ascertainment resulting from excess mortality in cattle near to disease onset. Ongoing levels of human exposure to BSE infectivity were also estimated, together with the effect on these of a range of possible exposure-reduction strategies that might replace the current rule banning tissue from cattle over 30 months (OTM) of age from the human food supply. While any policy change that allows a wider age range of animals into the human food supply will increase levels of human exposure to infectivity, the risk posed by such increases is small by comparison with historical exposure levels. Making the pessimistic assumption that there will be 5000 deaths during the variant Creutzfeldt-Jakob disease (vCJD) epidemic in total, our analysis indicates that replacement of the OTM rule with testing would result in 0.04 additional vCJD deaths over the next 60 years. However, there is substantial (more than 40-fold) uncertainty surrounding this estimate, the sources of which are discussed. PMID- 12908979 TI - Infection by the systemic fungus Epichloe glyceriae alters clonal growth of its grass host, Glyceria striata. AB - Parasites and pathogens are hypothesized to change host growth, reproduction and/or behaviour to increase their own transmission. However, studies which clearly demonstrate that parasites or pathogens are directly responsible for changes in hosts are lacking. We previously found that infection by the systemic fungus Epichloe glyceriae was associated with greater clonal growth by its host, Glyceria striata. Whether greater clonal growth resulted directly from pathogen infection or indirectly from increased likelihood of infection for host genotypes with greater clonal growth could not be determined because only naturally infected and uninfected plants were used. In this study, we decoupled infection and host genotype to evaluate the role of pathogen infection on host development and clonal growth. We found that total biomass production did not differ for clones of the same genotype, but infected clones allocated more biomass to clonal growth. Disinfected clones had more tillers and a greater proportion of their biomass in the mother ramet. Infected clones produced fewer tillers but significantly more and longer stolons than disinfected clones. These results support the hypothesis that pathogen infection directly alters host development. Parasite alteration of clonal growth patterns might be advantageous to the persistence and spread of host plants in some ecological conditions. PMID- 12908980 TI - How did pygmy shrews colonize Ireland? Clues from a phylogenetic analysis of mitochondrial cytochrome b sequences. AB - There is a long-standing debate as to how Ireland attained its present fauna; we help to inform this debate with a molecular study of one species. A 1110 base pair fragment of the mitochondrial cytochrome b gene was sequenced in 74 specimens of the pygmy shrew, Sorex minutus, collected from throughout its western Palaearctic range. Phylogenetic analysis of these sequences revealed several well-supported lineages. Most of the 65 haplotypes belonged to a northern lineage, which ranged from Britain in the west to Lake Baikal in the east. The other lineages were largely limited to Iberia, Italy and the Balkans. One exception, however, was a lineage found in both Ireland and Andorra. This affinity, and the large difference between the mitochondrial sequences of Irish and British individuals, suggest that pygmy shrews did not colonize Ireland via a land connection from Britain, as has been previously supposed, but instead were introduced by boat from southwest continental Europe. All the Irish pygmy shrews analysed were identical or very similar in cytochrome b sequence, suggesting an extreme founding event. PMID- 12908981 TI - Low propensity for aerial dispersal in specialist spiders from fragmented landscapes. AB - Aerial dispersal by ballooning is a passive flight, by which wind drag generates an upward lift on a silk thread. It is likely to reflect an aerial lottery, in which the absence of flight direction control is a serious cost for long-distance dispersal in a fragmented landscape. For species occurring in one patchily distributed habitat type, dispersal should evolve in a different way from morphological traits, directly linked to active dispersal. Therefore, we expect that if the risk of landing in an unsuitable habitat is lower than the probability of reaching a suitable habitat, selection should benefit a well developed ballooning behaviour. We investigated interspecific variation in the ballooning-initiating tiptoe behaviour as it is linked to spider dispersal performance. Our results indeed indicate that ballooning performance is negatively related to habitat specialization in spiders from patchy grey dunes, so habitat specialists are characterized by poorly developed dispersal behaviour. These findings are concordant with recent insights that dispersal is selected as risk spreading in generalists, while it is selected against in specialist species. PMID- 12908982 TI - Regulation of ants' foraging to resource productivity. AB - We investigate the behavioural rule used by ant societies to adjust their foraging response to the honeydew productivity of aphids. When a scout finds a single food source, the decision to lay a recruitment trail is an all-or-none response based on the opportunity for this scout to ingest a desired volume acting as a threshold. Here, we demonstrate, through experimental and theoretical approaches, the generic value of this recruitment rule that remains valid when ants have to forage on multiple small sugar feeders to reach their desired volume. Moreover, our experiments show that when ants decide to recruit nest mates they lay trail marks of equal intensity, whatever the number of food sources visited. A model based on the 'desired volume' rule of recruitment as well as on experimentally validated parameter values was built to investigate how ant societies adjust their foraging response to the honeydew productivity profile of aphids. Simulations predict that, with such recruiting rules, the percentage of recruiting ants is directly related to the total production of honeydew. Moreover, an optimal number of foragers exists that maximizes the strength of recruitment, this number being linearly related to the total production of honeydew by the aphid colony. The 'desired volume' recruitment rule that should be generic for all ant species is enough to explain how ants optimize trail recruitment and select aphid colonies or other liquid food resources according to their productivity profile. PMID- 12908983 TI - Insurance-based advantages for subordinate co-foundresses in a temperate paper wasp. AB - Recent explanations for the evolution of eusociality, focusing more on costs and benefits than relatedness, are largely untested. We validate one such model by showing that helpers in foundress groups of the paper wasp Polistes dominulus benefit from an insurance-based mechanism known as Assured Fitness Returns (AFRs). Experimental helper removals left remaining group members with more offspring than they would normally rear. Reduced groups succeeded in preserving the dead helpers' investment by rearing these extra offspring, even when helper removals occurred long before worker emergence. While helpers clearly gained from AFRs, offspring of lone foundresses failed after foundress death, so that AFRs represent a true advantage for helpers. Smaller, less valuable offspring were probably sacrificed to feed larger offspring, but reduced groups did not preferentially attract joiners or increase their foraging effort to compensate for their smaller workforce. We failed to detect a second insurance-based advantage, Survivorship Insurance, in which larger groups are less likely to fail than smaller groups. We suggest that through their use of small offspring as a food store to cope with temporary shortages, wasps may be less susceptible than vertebrates to offspring failure following the death of group members. PMID- 12908984 TI - Multiple mating and sequential mate choice in guppies: females trade up. AB - The trade-up hypothesis outlines a behavioural strategy that females could use to maximize the genetic benefits to their offspring. The hypothesis proposes that females should be more willing to accept a mate when the new male encountered is a superior genetic source to previous mates. We provide a direct test of the trade-up hypothesis using guppies (Poecilia reticulata), and evaluate both behavioural and paternity data. Virgin female guppies were presented sequentially with two males of varying attractiveness, and their responsiveness to each male was quantified. Male attractiveness (ornamentation) was scored as the amount of orange coloration on their body. Females were generally less responsive to second encountered males, yet responsiveness to second males was an increasing function of male ornamentation. These attractive second males also sired a greater proportion of the offspring. There was an overall tendency for last-male advantage in paternity, and this advantage was most exaggerated when the second male was more ornamented than the first. Finally, we found that our estimate of relative sperm number did not account for any significant variation in paternity. Our results suggest that female guppies may use pre-copulatory mechanisms to maximize the genetic quality of their offspring. PMID- 12908985 TI - The turn of the sword: length increases male swimming costs in swordtails. AB - Sexual selection via female mate choice can result in the evolution of elaborate male traits that incur substantial costs for males. Despite increased interest in how female mating preferences contribute to the evolution of male traits, few studies have directly quantified the locomotor costs of such traits. A sexually selected trait that could affect movement costs is the sword exhibited by male swordtail fishes: while longer swords may increase male mating success, they could negatively affect the hydrodynamic aspects of swimming activities. Here, we examine the energetic costs of the sword in Xiphophorus montezumae by experimentally manipulating sword length and measuring male aerobic metabolism during two types of activity, routine swimming and courtship swimming. Direct measurements of oxygen consumption indicate that males with longer swords expend more energy than males with shortened swords during both types of swimming. In addition, the sword increases the cost of male courtship. Thus, while sexual selection via female choice favours long swords, males with longer swords experience higher metabolic costs during swimming, suggesting that sexual and natural selection have opposing effects on sword evolution. This study demonstrates a hydrodynamic cost of a sexually selected trait. In addition, this study discriminates between the cost of a sexually selected trait used in courtship and other courtship costs. PMID- 12908986 TI - Optimal assessment of multiple cues. AB - In a wide range of contexts from mate choice to foraging, animals are required to discriminate between alternative options on the basis of multiple cues. How should they best assess such complex multicomponent stimuli? Here, we construct a model to investigate this problem, focusing on a simple case where a 'chooser' faces a discrimination task involving two cues. These cues vary in their accuracy and in how costly they are to assess. As an example, we consider a mate-choice situation where females choose between males of differing quality. Our model predicts the following: (i) females should become less choosy as the cost of finding new males increases; (ii) females should prioritize cues differently depending on how choosy they are; (iii) females may sometimes prioritize less accurate cues; and (iv) which cues are most important depends on the abundance of desirable mates. These predictions are testable in mate-choice experiments where the costs of choice can be manipulated. Our findings are applicable to other discrimination tasks besides mate choice, for example a predator's choice between palatable and unpalatable prey, or an altruist's choice between kin and non-kin. PMID- 12908987 TI - Divergent evolution during an experimental adaptive radiation. AB - How repeatable a process is evolution? Comparative studies of multicellular eukaryotes and experimental studies with unicellular prokaryotes document the repeated evolution of adaptive phenotypes during similar adaptive radiations, suggesting that the outcome of adaptive radiation is broadly reproducible. The goal of this study was to test this hypothesis by using phenotypic traits to infer the genetic basis of adaptation to simple carbon-limited environments in an extensive adaptive radiation. We used a clone of the bacterium Pseudomonas fluorescens to found two sets of experimental lines. The first set of lines was allowed to adapt to one of 23 novel environments for 1100 generations while the second set of lines was allowed to accumulate mutations by drift for 2000 generations. All lines were then assayed in the 95 environments provided by Biolog microplates to determine the phenotypic consequences of selection and drift. Replicate selection lines propagated in a common environment evolved similar adaptive components of their phenotype but showed extensive variation in non-adaptive phenotypic traits. This variation in non-adaptive phenotypic traits primarily resulted from the ascendance of different beneficial mutations in different lines. We argue that these results reconcile experimental and comparative approaches to studying adaptation by demonstrating that the convergent phenotypic evolution that occurs during adaptive radiation may be associated with radically different sets of beneficial mutations. PMID- 12908988 TI - Re-visioning refugee health: the Victorian Immigrant Health Programme. AB - Few published data are available to support the development and management of health services for refugees in developed countries, despite evidence that refugees are at high risk of suffering a wide range of both physical and mental health problems. Meeting the health needs of increasing numbers of refugees worldwide in culturally acceptable and financially feasible ways is a key challenge to our healthcare systems. This paper reports on seven platforms identified in the development of the Victorian Immigrant Health Programme (VIHP) that the VIHP believes should underpin the management and delivery of healthcare to newly arrived refugees. Sharing this information facilitates and promotes essential collaborative work in the field of refugee health, and assists to prevent duplicate efforts, given constraints on resources for refugee healthcare provision in Australia and elsewhere. PMID- 12908989 TI - Top-rated British business research: has the emperor got any clothes? AB - Business schools have great prestige and charge large amounts of money for their courses. But how good is the science on which they base their prescriptions for action? To find out we examined the published output from the only three British business schools with the highest (5*) research assessment ranking at the time the articles were published. We conclude that theory development and model construction are often elegant. However, the methods used to obtain primary empirical information to confirm or refute the theories or populate models are poor, at least from a positivist or pragmatic ontological perspective. Large scale comparative studies made up only a small proportion of research output from the business schools. Literature reviews were not systematic. The sampling frame and rationale for selection of cases for study are inadequately described. The methods of data collection were frequently not given in sufficient detail to enable the study to be replicated and the conclusions tended to go far beyond what the data by themselves could support. However, this does not have to be the case-there are excellent examples of research in social sciences. We conclude, therefore, that top-rated British business research is a scantily clad emperor. PMID- 12908990 TI - A province-wide study of the association between hospital resource allocation and length of stay. AB - The relationship between hospital resource allocation and clinical efficiency is poorly understood. Within the single-payer healthcare system in Ontario, Canada, the association between hospital spending patterns and length of stay was studied using data from 1117090 patient discharges in 1997/8 at 162 of 171 acute care hospitals. A weighted regression model was created using an overall hospital length of stay index (actual length of stay divided by predicted length of stay) as the dependent variable. Control variables included: hospital size, teaching activity, occupancy rate, rural location and geographic region. Four independent spending variables were defined as a percentage of total hospital spending: nursing, ambulatory care, administration and support, and diagnostics and therapeutics. The reduced regression model had an r-squared of 0.45. Across all spending variables, hospitals spending relatively too little or too much had significantly longer length of stay. Hospitals' overall pattern of resource allocation was also significantly associated with length of stay. Thus, measurable clinical effects can be seen with resource allocation decisions made by hospital management, supporting the need for rigorous decision-making processes. Future research should focus on exploring the nature of this relationship and the potential interdependencies among hospital services that cause this effect. PMID- 12908991 TI - Adoption of regulatory compliance programmes across United States healthcare organizations: a view of institutional disobedience. AB - The growing acceptance of evidence-based decision-support systems in healthcare organizations has resulted in recognition of information accuracy as a key area of organizational management. In the United States, rigid data mandates related to information management have met with some resistance from healthcare provider groups, who have traditionally found little relevance between personalized healthcare practice and accurate information. Variation in management practice poses quality problems in such an environment, since it precludes comparisons across larger markets or areas, a critical component of evidence-based quality assessments. In this study, a national census of health information managers was employed to provide a benchmark of the degree of such variation, examining how proper billing compliance practices vary across organization types as well as market area indicators. Findings here suggest that managers continue to ignore, to some extent, regulatory compliance standards, despite nationwide laws that mandate adoption of uniform compliance practices and programmes. The level of adoption of compliance management in this study varied significantly across practice characteristics and areas, suggesting the existence of barriers to cross market comparative performance assessment. PMID- 12908992 TI - Gaining and maintaining commitment to large-scale change in healthcare organizations. AB - Healthcare administrators have sought to improve the quality of healthcare services by using organizational change as a lever. Unfortunately, evaluations of organizational change efforts in areas such as total quality management (TQM), continuous quality improvement (CQI), and organizational restructuring have indicated that these change programmes have not fulfilled their promise in improving service delivery. Furthermore, there are no easy answers as to why so many large-scale change programmes are unsuccessful. The aim of this analysis is to provide insights into practices that may be utilized to improve the chances of successful change management. It is proposed that in order to effect change, implementers must first gain commitment to the change. This is done by ensuring organizational readiness for change, surfacing dissatisfaction with the present state, communicating a clear vision of the proposed change, promoting participation in the change effort, and developing a clear and consistent communication plan. However gaining commitment is not enough. Many change programmes have been initially perceived as being successful but long-term success has been elusive. Therefore, maintaining commitment during the uncertainty associated with the transition period is imperative. This can be done by successfully managing the transition using action steps such as consolidating change using feedback mechanisms and making the change a permanent part of the organization's culture. PMID- 12908993 TI - The tendering process: flaws and all. AB - The tendering process has become the dominant and now traditional approach to the allocation of research and consultancy projects. This is largely built around the notion that it ensures accountability and probity within the allocative procedure, and that high quality of work and 'value for money' are the outcomes. It appears to have become an institutionalized and unchallengeable process. Yet it is rarely costed in terms of the true resource implications for all the organizations involved, nor is quality assessed in terms of alternative processes that might be employed. This paper explores the tendering process in terms of factors that may suggest that the quality of work produced via the tendering process is not always as good as it might be. Also in a hypothetical example involving the university sector and the National Health Service, data are presented indicating that the overall cost to the public sector is often actually greater than the value of the contract being allocated. An alternative preferred provider relationship-based allocative process is advocated as an improvement to the current established tendering procedure. PMID- 12908994 TI - Break-even analysis revisited: the need to adjust for profitability, the collection rate and autonomous income. AB - This paper modifies traditional break-even analysis and develops a model that reflects the influence of variation in payer mix, the collection rate, profitability and autonomous income on the desired volume alternative. The augmented model indicates that a failure to adjust for uncollectibles and the net surplus results in a systematic understatement of the desired volume alternative. Conversely, a failure to adjust for autonomous income derived from the operation of cafeterias, gift shops or an organization's investment in marketable securities produces an overstatement of the desired volume. In addition, this paper uses Microsoft Excel to develop a spreadsheet that constructs a pro forma income statement, expressed in terms of the contribution margin. The spreadsheet also relies on the percentage of sales or revenue approach to prepare a balance sheet from which indicators of fiscal performance are calculated. Hence, the analysis enables the organization to perform a sensitivity analysis of potential changes in the desired volume, the operating margin, the current ratio, the debt: equity ratio and the amount of cash derived from operations that are associated with expected variation in payer mix, the collection rate, grouped by payer, the net surplus and autonomous income. PMID- 12908995 TI - Constructing management practice in the new public management: the case of mental health managers. AB - There has been a growth in managerialism of professionals in the Public Sector. This paper contributes to this debate by examining perceptions of senior managers in mental health services within Health Authorities, NHS Trusts and Local Authorities' Social Services departments in Wales. The paper explores how different professions responded differently to the growth of managerialism. The results suggest important differences and similarities between management in health and social services settings. PMID- 12908996 TI - Summary of the NHLBI Working Group on Research on Hypertension During Pregnancy. AB - A Working Group on Research in Hypertension in Pregnancy was recently convened by the National Heart Lung and Blood Institute to determine the state of knowledge in this area and suggest appropriate directions for research. Hypertensive disorders in pregnancy, especially preeclampsia, are a leading cause of maternal mortality worldwide and even in developed countries increase perinatal mortality five-fold. Much has been learned about preeclampsia but gaps in the knowledge necessary to direct therapeutic strategies remain. Oxidative stress is a biologically plausible contributor to the disorder that may be amenable to intervention. Hypertension that antedates pregnancy (chronic hypertension) bears many similarities to hypertension in nonpregnant women but the special setting of pregnancy demands information to guide evidence based therapy. The recommendations of the Working Group are to attempt a clinical trial of antioxidant therapy to prevent preeclampsia that is be complemented by mechanistic research to increase understanding of the genetics and pathogenesis of the disorder. For chronic hypertension clinical trials are recommended to direct choice of drugs, evaluate degree of control and assess implications to the mother and fetus. Recommendations to increase participation in this research are also presented. PMID- 12908997 TI - Neutrophil activation and C-reactive protein concentration in preeclampsia. AB - Preeclamptic pregnancies seem to be associated with a higher extent of inflammation compared with normal ones. We intended to test this proposal and also to clarify the contribution of some variables in such inflammatory process. We measured total and differential leukocyte count, serum C-reactive protein (CRP), and plasma levels of lactoferrin, elastase, and granulocyte-macrophage colony-stimulating factor (GM-CSF). Uric acid was also evaluated and used as an indicator of the severity of the disease. A cross-sectional study was performed by evaluating healthy and preeclamptic women in the third trimester of gestation (n = 67 and n = 51, respectively) and 24 to 48 h postpartum (n = 32 and n = 26, respectively). When comparing the third trimester of normal and preeclamptic pregnancies, we found significantly higher levels of uric acid, CRP, and elastase, and a significantly higher elastase to neutrophil ratio in the pathologic group. However, for CRP, statistical significance was lost after adjustment for maternal weight. No significant differences were found in total leukocyte count, plasma levels of GM-CSF, and lactoferrin between groups. In preeclampsia, a significant positive correlation was found between elastase and lactoferrin and these neutrophil activation products correlated positively with uric acid level. Considering the analysis of all variables in the postpartum period, only CRP and uric acid levels were significantly elevated in the pathologic group. However, CRP differences obtained in the puerperium seem to be influenced by the increased number of dystocic deliveries in the preeclamptic group. In conclusion, our data suggest that inflammation is further pronounced in preeclampsia and that the extent of neutrophil activation correlates with the severity of this syndrome. PMID- 12908998 TI - Subclassification of preeclampsia. AB - Preeclampsia is a heterogeneous disorder, and as with other diseases (e.g., type I and type II diabetes), progress in the understanding of this disorder would be assisted greatly if subtypes could be characterized. We suggest that a first step would be to subdivide preeclampsia into early-onset disease (< 34 + 0 weeks') and late onset disease (> 34 + 0 weeks'). PMID- 12908999 TI - Does endothelial nitric oxide synthase gene variation play a role in the occurrence of hypertension in pregnancy? AB - OBJECTIVE: Nitric oxide is suggested to play a role in the development of preeclampsia. METHODS: We studied 61 patients with gestational hypertension (GH), 77 with GH and significant proteinuria (urine protein excretion > or = 300 mg/24 h), 82 with essential hypertension (EH) and 188 normotensive women with at least one normal pregnancy. MAIN OUTCOME MEASURE(S): A polymorphism within the constitutive endothelial nitric oxide synthase (ecNOS) gene in various types of hypertension in pregnancy was explored. RESULTS: Allelic and genotypic frequencies did not differ between controls and case groups. A significant difference was observed between the frequency of the rare allele in GH patients and that in EH group (chi2: 4.47, P <.04). This difference approximated the significance when GH subjects with or without proteinuria were grouped (chi2 square: 3.33; P =.068). Cigarette smoking or gravidity did not interact with the ecNOS polymorphism in identifying different types of hypertension in this setting. CONCLUSION: Our findings argue against an association between ecNOS polymorphism and preeclampsia and support the hypothesis for a different pathogenesis of GH in respect to EH. PMID- 12909000 TI - Fetal DNA quantitation in peripheral blood is not useful as a marker of disease severity in women with preeclampsia. AB - OBJECTIVE: Trophoblast migration into the maternal circulation is increased in preeclampsia and can trigger the endothelial dysfunction that characterizes the clinical disease. We hypothesised that 'fetal trafficking' is increased in women with severe preeclampsia and that the quantity of trafficking is greater in women with more severe disease. STUDY DESIGN: To test the hypothesis, we used a technique that quantifies a genetic marker specific for the fetomaternal unit, that is, the SRY gene in a pregnant woman carrying a male fetus. Thirty two women with pre-eclampsia and 32 control women (women without preeclampsia) were recruited. Preeclampsia was defined according to the International Society for the Study of Hypertension in Pregnancy (ISSHP). All subjects with preeclampsia had evidence of the multisystemic nature of the disease. DNA was extracted from maternal peripheral blood and RTQ PCR analysis was performed to quantify the fetal DNA (SRY) and the total DNA (beta-actin) in each sample. The ratio of fetal to total DNA was calculated and compared between women with preeclampsia and controls. RESULTS: The women with preeclampsia and the control women did not differ in parity, blood pressure at booking, and gestational age at sampling. The groups differed significantly in age (29 +/- 5.7 vs 25 +/- 5.1 years; P =.007), diastolic blood pressure (DBP) at sampling (101 +/- 9.5 vs 70 +/- 5.5 mm Hg; P <.0001), gestational age at delivery (33 +/- 4.3 vs 39 +/- 1.8 weeks; P <.001), and fetal weight (1.98 +/- 1 vs 3.35 +/- 0.5 kgs; P <.0001). SRY was detected in 31 out of 32 women with preeclampsia and in 24 out of 32 control women (P <.001). The median SRY copy number per micro L was greater in women with preeclampsia (10.6, interquartile range 12.89) than in the control women (8.6, interquartile range 20.1) but these differences were not statistically significant at P =.75. The median ratio of fetal to total DNA was almost identical in both groups (0.06, interquartile range 0.13) in PET compared to (0.06, interquartile range 0.17) the control women. No correlation was found between the quantity of fetal DNA and disease severity. CONCLUSION: Fetal trafficking is more likely to be detected in women with preeclampsia compared to control women but the quantity does not appear to correlate with disease severity. PMID- 12909001 TI - Surface expression of neutrophil adhesion molecules in pregnant women at risk for hypertensive complications. AB - OBJECTIVE: To determine if neutrophil activation is a pathogenetic factor in hypertensive disorders in pregnancy, the neutrophil expression of adhesion molecules was prospectively investigated in pregnant women at risk, prior to the development of hypertensive complications. METHODS: Two neutrophil activation parameters, beta2-integrin (CD11b) and l-selectin (CD62L), were assessed at admission between 14 and 24 weeks of gestation in 82 pregnant women at risk of preeclampsia and other hypertensive complications. Results were compared to those in 20 healthy normotensive women. RESULTS: Of the 82 women at risk, 23 (28%) developed hypertensive complications: 9 (11%) preeclampsia and 14 (17%) others, such as intrauterine growth restriction (n = 6), fetal or neonatal loss (n = 8), and preterm delivery (< or = 30 weeks of gestation) (n = 8). All pregnancy outcome measures were significantly worse in the patients with complications than in those at risk but without complications or the healthy controls. Expression of beta2-integrin was significantly higher in early stages of pregnancy in the women who eventually developed complications than the women who did not, P =.019, or the healthy controls, P =.049. CONCLUSIONS: Surface expression of beta2-integrin is increased in pregnant women at risk for hypertensive complications before the clinical manifestations of the disorder. PMID- 12909002 TI - The receptor for advanced glycation end products (RAGE) is elevated in women with preeclampsia. AB - A leading theory of the pathophysiology of preeclampsia is that oxidative stress induces vascular endothelial cell dysfunction. Advanced glycation end products (AGEs) form when aldose sugars react nonenzymatically with proteins under conditions of oxidative stress. AGEs are circulating molecules and can generate reactive oxygen species and vascular dysfunction (in diabetes and atherosclerosis) through an association with cell surface receptors (RAGE). RAGE is a multiligand receptor, expressed in vascular tissue, which is upregulated by its own ligands. Insulin resistance and obesity are risk factors for developing preeclampsia, as well as being conditions that would increase RAGE levels. Thus, we hypothesized that women with preeclampsia will have elevated levels of RAGE protein compared with normal pregnant women. Biopsies of nonlaboring myometrium as well as omentum were taken from normal pregnant and preeclamptic women. Nonpregnant samples were obtained at the time of hysterectomy. Tissue sections were immunostained with anti-RAGE as well as anti-alpha-actin and anti-von Willebrand factor (to identify blood vessels and intact endothelial cells). Staining intensity was qualitatively described as well as given an intensity score, with the identity of the section concealed. Nonpregnant myometrial and omental vessels showed very low to undetectable levels of RAGE staining. Pregnancy induced a significant increase in RAGE protein levels in both myometrium and omental vasculature. Blood vessels from women with preeclampsia consistently had intense staining for RAGE in both vessel beds. Thus, our data suggest that since RAGE activation can induce similar pathophysiologic changes to those observed in women with preeclampsia (including NFkappaB activation, increased TNFalpha and endothelin), elevated RAGE protein may be contributing to the vascular dysfunction in preeclampsia. PMID- 12909003 TI - The effect of treatment on endothelin-1 concentration and mean arterial pressure in preeclampsia and eclampsia. AB - OBJECTIVE: The aim of this study was to determine the effect of treatment on endotelin-1 concentration that is considered to have role in etiopathogenesis of eclampsia and preeclampsia. METHODS: Nine patients with eclampsia and 12 patients with preeclampsia were included to the study. Endothelin-1 levels were measured before and after magnesium sulfate treatment. After the magnesium sulfate administration, if the blood pressure was still elevated, nifedipine was given. RESULTS: Endothelin-1 levels of preeclamptic patients before and after magnesium sulfate treatment were 16.9 +/- 2.3 fmol/ml and 14.6 +/- 1.9 fmol/ml respectively (p < 0.05). The same measurements of eclamptic patients were 18.1 +/- 3.2 and 14.7 +/- 3.4 respectively (p < 0.05). The mean blood pressures of preeclamptic patients before and after magnesium sulfate were 125.8 +/- 7.3 mm-Hg, 118.2 +/- 8.7 mm-Hg respectively, and the same measurements of eclamptic patients were 136.0 +/- 12.4 mm-Hg and 123.1 +/- 10.6 mm-Hg respectively (p < 0.05, p < 0.05). CONCLUSION: Treatment had been found to have negative effects on endothelin-1 levels that is considered to play an important role on etiopathogenesis of eclampsia and preeclampsia. PMID- 12909004 TI - Expression of vascular endothelial growth factor in peripheral blood cells of preeclamptic women. AB - OBJECTIVE: Preeclampsia is associated with platelet and endothelial dysfunction. Vascular endothelial growth factor (VEGF) is found in peripheral blood leukocytes and released from platelets on activation. We analyzed the content of (VEGF) in peripheral blood cells of preeclamptic women. METHODS: The VEGF content of platelets, mononuclear white blood cells, and granulocytes of peripheral blood were analyzed from 12 women with preeclampsia, 19 healthy pregnant women, and 20 nonpregnant women. Protein released from lysed cells was analyzed by enzyme linked immunoassay (ELISA). RESULTS: Platelet VEGF content of preeclamptic women (0.081 ng/109 cells, 0.016 to 2.7 ng/109 cells; median, range) was similar to that of healthy pregnant women (0.31 ng/109 cells, 0.013 to 0.92 ng/109 cells) and that of nonpregnant (0.073 ng/109 cells, 0.012 to 0.76 ng/109 cells) women. Likewise, the VEGF content of granulocytes was similar in preeclamptic (18.5 ng/109 cells, 1.2 to 193 ng/109 cells), healthy pregnant (25.3 ng/109 cells, 0.8 to 441 ng/109 cells), and nonpregnant (29 ng/109 cells, 0.25 to 200 ng/109 cells) women. In mononuclear cells, the VEGF content of healthy pregnant women was higher (4.4 ng/109 cells, 0.13 to 13.7 ng/109 cells) than in nonpregnant women (1.7 ng/109 cells, 0.15 to 11.4 ng/109 cells, P < 0.05). Also, the mononuclear cell VEGF content of preeclamptic women (8.2 ng/109 cells, 0.04 to 23 ng/109 cells) tended to be higher than in nonpregnant women (P approximately 0.07). CONCLUSION: Uncomplicated pregnancy is associated with an elevated VEGF content of mononuclear cells. Preeclampsia does not seem to affect the VEGF content of maternal peripheral blood mononuclear cells, granulocytes, or platelets. PMID- 12909005 TI - Severe maternal morbidity associated with hypertensive disorders in pregnancy in the United States. AB - OBJECTIVES: This study was to report the incidence of severe maternal morbidity associated with hypertensive disorders of pregnancy in the United States. STUDY DESIGN: We used data from the National Hospital Discharge Survey, a nationally representative sample of discharge records, from 1988 to 1997. The database consisted of approximately 300,000 deliveries, which represented 39 million births during the 10-year period. RESULTS: The overall incidence of hypertensive disorders in pregnancy was 5.9% [95% confidence interval (CI): 5.2 to 6.5%]. Eclampsia was reported at 1.0 per 1,000 deliveries (95% CI: 0.8 to 1.2). The incidence of eclampsia, severe preeclampsia, and superimposed preeclampsia remained unchanged during the 10-year period. Women with preeclampsia and eclampsia had a 3- to 25-fold increased risk of severe complications, such as abruptio placentae, thrombocytopenia, disseminated intravascular coagulation, pulmonary edema, and aspiration pneumonia. More than half of women with preeclampsia and eclampsia had cesarean delivery. African American women not only had higher incidence of hypertensive disorders in pregnancy but also tended to have a greater risk for most severe complications. Preeclamptic and eclamptic women younger than 20 years or older than 35 years had substantially higher morbidity. CONCLUSION: Preeclampsia and eclampsia carry a high risk for severe maternal morbidity. Compared to Caucasians, African Americans have higher incidence of hypertensive disorders in pregnancy and suffer from more severe complications. PMID- 12909006 TI - Translational standby sites: how ribosomes may deal with the rapid folding kinetics of mRNA. AB - We have previously shown that stable base-pairing at a translational initiation site in Escherichia coli can inhibit translation by competing with the binding of ribosomes. When the base-pairing is not too strong, this competition is won by the ribosomes, resulting in efficient translation from a structured ribosome binding site (RBS). We now re-examine these results in the light of RNA folding kinetics and find that the window during which a folded RBS is open is generally much too short to recruit a 30S ribosomal subunit from the cytoplasm. We argue that to achieve efficient expression, a 30S subunit must already be in contact with the mRNA while this is still folded, to shift into place as soon as the structure opens. Single-stranded regions flanking the structure may constitute a standby site, to which the 30S subunit can attach non-specifically. We propose a steady-state kinetic model for the early steps of translational initiation and use this to examine various quantitative aspects of standby binding. The kinetic model provides an explanation of why the earlier equilibrium competition model predicted implausibly high 30S-mRNA affinities. Because all RNA is structured to some degree, standby binding is probably a general feature of translational initiation. PMID- 12909007 TI - Stop codons and UGG promote efficient binding of the polypeptide release factor eRF1 to the ribosomal A site. AB - To investigate the codon dependence of human eRF1 binding to the mRNA-ribosome complex, we examined the formation of photocrosslinks between ribosomal components and mRNAs bearing a photoactivable 4-thiouridine probe in the first position of the codon located in the A site. Addition of eRF1 to the phased mRNA ribosome complexes triggers a codon-dependent quenching of crosslink formation. The concentration of eRF1 triggering half quenching ranges from low for the three stop codons, to intermediate for s4UGG and high for other near-cognate triplets. A theoretical analysis of the photochemical processes occurring in a two-state bimolecular model raises a number of stringent conditions, fulfilled by the system studied here, and shows that in any case sound KD values can be extracted if the ratio mT/KD<<1 (mT is total concentration of mRNA added). Considering the KD values obtained for the stop, s4UGG and sense codons (approximately 0.06 microM, 0.45 microM and 2.3 microM, respectively) and our previous finding that only the stop and s4UGG codons are able to promote formation of an eRF1-mRNA crosslink, implying a role for the NIKS loop at the tip of the N domain, we propose a two-step model for eRF1 binding to the A site: a codon-independent bimolecular step is followed by an isomerisation step observed solely with stop and s4UGG codons. Full recognition of the stop codons by the N domain of eRF1 triggers a rearrangement of bound eRF1 from an open to a closed conformation, allowing the universally conserved GGQ loop at the tip of the M domain to come into close proximity of the peptidyl transferase center of the ribosome. UGG is expected to behave as a cryptic stop codon, which, owing to imperfect eRF1-codon recognition, does not allow full reorientation of the M domain of eRF1. As far as the physical steps of eRF1 binding to the ribosome are considered, they appear to closely mimic the behaviour of the tRNA/EF-Tu/GTP complex, but clearly eRF1 is endowed with a greater conformational flexibility than tRNA. PMID- 12909008 TI - Structure of the pyrimidine-rich internal loop in the poliovirus 3'-UTR: the importance of maintaining pseudo-2-fold symmetry in RNA helices containing two adjacent non-canonical base-pairs. AB - Formation of non-canonical base-pairs in RNA often plays a very important functional role. In addition they frequently serve as factors in stabilizing the secondary structure elements that provide the frame of large compact RNA structures. Here we describe the structure of an internal loop containing a 5'CU3'/5'UU3' non-canonical tandem base-pair motif, which is conserved within the 3'-UTR of poliovirus-like enteroviruses. Structural details reveal striking regularities of the local helix geometry, resulting from alternating geometrical adjustments, which are important for understanding and predicting stabilities and configurations of tandem non-canonical base-pairs. The C-U and U-U base-pairs severely contract the minor groove of the sugar-phosphate backbone, which might be important for protein recognition or binding to other RNA elements. PMID- 12909009 TI - Different genome maintenance strategies in human and tobacco cells. AB - In this work, genome maintenance strategies of organisms belonging to different kingdoms (animals versus plants) but of similar genome size were investigated using a novel, universal double-strand break (DSB) repair assay. Different plasmids linearised with KpnI, Acc65I or EcoRV yielding either 3' or 5' protruding or blunt DNA termini, respectively, were transfected into HeLa cells and Nicotiana plumbaginifolia protoplasts and assayed for the efficiency and fidelity of DSB repair. We show that the mechanism of break sealing is similar but that drastic differences are seen in the fidelity of repair: in HeLa cells, 50-55% DSBs were repaired precisely, compared to as little as 15-30% in tobacco cells. Moreover, the DSB repair in plants resulted in 30-40% longer deletions and significantly shorter insertions. Combined, these led to more than twofold larger net DNA loss in tobacco cells. Our observations point to possible differences in the strategies of DSB repair and genome maintenance in plants and animals. PMID- 12909010 TI - A conserved insertion in protein-primed DNA polymerases is involved in primer terminus stabilisation. AB - Protein-primed DNA polymerases form a subgroup of the eukaryotic-type DNA polymerases family, also called family B or alpha-like. A multiple amino acid sequence alignment of this subgroup of DNA polymerases led to the identification of two insertions, TPR-1 and TPR-2, in the polymerisation domain. We showed previously that Asp332 of the TPR-1 insertion of phi29 DNA polymerase is involved in the correct orientation of the terminal protein (TP) for the initiation of replication. In this work, the functional role of two other conserved residues from TPR-1, Lys305 and Tyr315, has been analysed. The four mutant derivatives constructed, K305I, K305R, Y315A and Y315F, displayed a wild-type 3'-5' exonuclease activity on single-stranded DNA. However, when assayed on double stranded DNA such activity was higher than that of the wild-type enzyme. This activity led to a reduced pol/exo ratio, suggesting a defect in stabilising the primer terminus at the polymerase active site. On the other hand, although mutant polymerases K305I and Y315A were able to couple processive DNA polymerisation to strand displacement, they were severely impaired in phi29 TP-DNA replication. The possible role of the TPR-1 insertion in the set of interactions with the nascent chain during the first steps of TP-DNA replication is discussed. PMID- 12909011 TI - DNA binding of a non-sequence-specific HMG-D protein is entropy driven with a substantial non-electrostatic contribution. AB - The thermal properties of two forms of the Drosophila melanogaster HMG-D protein, with and without its highly basic 26 residue C-terminal tail (D100 and D74) and the thermodynamics of their non-sequence-specific interaction with linear DNA duplexes were studied using scanning and titration microcalorimetry, spectropolarimetry, fluorescence anisotropy and FRET techniques at different temperatures and salt concentrations. It was shown that the C-terminal tail of D100 is unfolded at all temperatures, whilst the state of the globular part depends on temperature in a rather complex way, being completely folded only at temperatures close to 0 degrees C and unfolding with significant heat absorption at temperatures below those of the gross denaturational changes. The association constant and thus Gibbs energy of binding for D100 is much greater than for D74 but the enthalpies of their association are similar and are large and positive, i.e. DNA binding is a completely entropy-driven process. The positive entropy of association is due to release of counterions and dehydration upon forming the protein/DNA complex. Ionic strength variation showed that electrostatic interactions play an important but not exclusive role in the DNA binding of the globular part of this non-sequence-specific protein, whilst binding of the positively charged C-terminal tail of D100 is almost completely electrostatic in origin. This interaction with the negative charges of the DNA phosphate groups significantly enhances the DNA bending. An important feature of the non-sequence specific association of these HMG boxes with DNA is that the binding enthalpy is significantly more positive than for the sequence-specific association of the HMG box from Sox-5, despite the fact that these proteins bend the DNA duplex to a similar extent. This difference shows that the enthalpy of dehydration of apolar groups at the HMG-D/DNA interface is not fully compensated by the energy of van der Waals interactions between these groups, i.e. the packing density at the interface must be lower than for the sequence-specific Sox-5 HMG box. PMID- 12909012 TI - Coactivator binding promotes the specific interaction between ligand and the pregnane X receptor. AB - The pregnane X receptor (PXR) detects the presence of a wide variety of endogenous and xenobiotic compounds, and is a master regulator of the expression of genes central to drug metabolism and excretion. We present the 2.0A crystal structure of the human PXR ligand-binding domain (LBD) in complex with the cholesterol-lowering compound SR12813 and a 25 amino acid residue fragment of the human steroid receptor coactivator-1 (SRC-1) containing one LXXLL motif. PXR crystallizes as a homodimer in the asymmetric unit in this structure and possesses a novel alpha2 helix adjacent to its ligand-binding cavity. The SRC-1 peptide forms two distinct helices and binds adjacent to the ligand-dependent transactivation AF-2 helix on the surface of PXR. In contrast with previous PXR structures, in which SR12813 bound in multiple orientations, the small SR12813 agonist in this structure binds in a single, unique orientation within the receptor's ligand-binding pocket and contacts the AF-2 helix. Thermal denaturation studies reveal that the SR12813 ligand and SRC-1 coactivator peptide each stabilize the LBD of PXR, and that together they exert an additive effect on the stability of the receptor. These results indicate that the binding of coactivator to the surface of PXR limits the ability of this promiscuous receptor to "breathe" and helps to trap a single, active conformation of SR12813. They further reveal that specificity is required for PXR activation. PMID- 12909013 TI - Catalysing new reactions during evolution: economy of residues and mechanism. AB - The diversity of function in some enzyme superfamilies shows that during evolution, enzymes have evolved to catalyse different reactions on the same structure scaffold. In this analysis, we examine in detail how enzymes can modify their chemistry, through a comparison of the catalytic residues and mechanisms in 27 pairs of homologous enzymes of totally different functions. We find that evolution is very economical. Enzymes retain structurally conserved residues to aid catalysis, including residues that bind catalytic metal ions and modulate cofactor chemistry. We examine the conservation of residue type and residue function in these structurally conserved residue pairs. Additionally, enzymes often retain common mechanistic steps catalyzed by structurally conserved residues. We have examined these steps in the context of their overall reactions. PMID- 12909014 TI - Structural basis of carbohydrate recognition by the lectin LecB from Pseudomonas aeruginosa. AB - The crystal structure of Pseudomonas aeruginosa fucose-specific lectin LecB was determined in its metal-bound and metal-free state as well as in complex with fucose, mannose and fructopyranose. All three monosaccharides bind isosterically via direct interactions with two calcium ions as well as direct hydrogen bonds with several side-chains. The higher affinity for fucose is explained by the details of the binding site around C6 and O1 of fucose. In the mannose and fructose complexes, a carboxylate oxygen atom and one or two hydroxyl groups are partly shielded from solvent upon sugar binding, preventing them from completely fulfilling their hydrogen bonding potential. In the fucose complex, no such defects are observed. Instead, C6 makes favourable interactions with a small hydrophobic patch. Upon demetallization, the C terminus as well as the otherwise rigid metal-binding loop become more mobile and adopt multiple conformations. PMID- 12909015 TI - Crystal structure of an ADP-dependent glucokinase from Pyrococcus furiosus: implications for a sugar-induced conformational change in ADP-dependent kinase. AB - ADP-dependent kinases are used in the modified Embden-Meyerhoff pathway of certain archaea. Our previous study has revealed a mechanism for ADP-dependent phosphoryl transfer by Thermococcus litoralis glucokinase (tlGK), and its evolutionary relationship with ATP-dependent ribokinases and adenosine kinases (PFKB carbohydrate kinase family members). Here, we report the crystal structure of glucokinase from Pyrococcus furiosus (pfGK) in a closed conformation complexed with glucose and AMP at 1.9A resolution. In comparison with the tlGK structure, the pfGK structure shows significant conformational changes in the small domain and a region around the hinge, suggesting glucose-induced domain closing. A part of the large domain next to the hinge is also shifted accompanied with domain closing. In the pfGK structure, glucose binds in a groove between the large and small domains, and the electron density of O1 atoms for both the alpha and beta anomer configurations was observed. The structural details of the sugar-binding site of ADP-dependent glucokinase were firstly clarified and then site-directed mutagenesis analysis clarified the catalytic residues for ADP-dependent kinase, such as Arg205 and Asp451 of tlGK. Homology search and multiple alignment of amino acid sequences using the information obtained from the structures reveals that eucaryotic hypothetical proteins homologous to ADP-dependent kinases retain the residues for the recognition of a glucose substrate. PMID- 12909016 TI - Structural basis for recognition and catalysis by the bifunctional dCTP deaminase and dUTPase from Methanococcus jannaschii. AB - Potentially mutagenic uracil-containing nucleotide intermediates are generated by deamination of dCTP, either spontaneously or enzymatically as the first step in the conversion of dCTP to dTTP. dUTPases convert dUTP to dUMP, thus avoiding the misincorporation of dUTP into DNA and creating the substrate for the next enzyme in the dTTP synthetic pathway, thymidylate synthase. Although dCTP deaminase and dUTPase activities are usually found in separate but homologous enzymes, the hyperthermophile Methanococcus jannaschii has an enzyme, DCD-DUT, that harbors both dCTP deaminase and dUTP pyrophosphatase activities. DCD-DUT has highest activity on dCTP, followed by dUTP, and dTTP inhibits both the deaminase and pyrophosphatase activities. To help clarify structure-function relationships for DCD-DUT, we have determined the crystal structure of the wild-type DCD-DUT protein in its apo form to 1.42A and structures of DCD-DUT in complex with dCTP and dUTP to resolutions of 1.77A and 2.10A, respectively. To gain insights into substrate interactions, we complemented analyses of the experimentally defined weak density for nucleotides with automated docking experiments using dCTP, dUTP, and dTTP. DCD-DUT is a hexamer, unlike the homologous dUTPases, and its subunits contain several insertions and substitutions different from the dUTPase beta barrel core that likely contribute to dCTP specificity and deamination. These first structures of a dCTP deaminase reveal a probable role for an unstructured C terminal region different from that of the dUTPases and possible mechanisms for both bifunctional enzyme activity and feedback inhibition by dTTP. PMID- 12909017 TI - The structural basis of receptor-binding by Escherichia coli associated with diarrhea and septicemia. AB - GafD in Escherichia coli G (F17) fimbriae is associated with diarrheal disease, and the structure of the ligand-binding domain, GafD1-178, has been determined at 1.7A resolution in the presence of the receptor sugar N-acetyl-D-glucosamine. The overall fold is a beta-barrel jelly-roll fold. The ligand-binding site was identified and localized to the side of the molecule. Receptor binding is mediated by side-chain as well main-chain interactions. Ala43-Asn44, Ser116 Thr117 form the sugar acetamide specificity pocket, while Asp88 confers tight binding and Trp109 appears to position the ligand. There is a disulfide bond that rigidifies the acetamide specificity pocket. The three fimbrial lectins, GafD, FimH and PapG share similar beta-barrel folds but display different ligand binding regions and disulfide-bond patterns. We suggest an evolutionary path for the evolution of the very diverse fimbrial lectins from a common ancestral fold. PMID- 12909018 TI - Thermotoga maritima IscU. Structural characterization and dynamics of a new class of metallochaperone. AB - Members of the IscU family of proteins are among the most conserved of all protein groups, extending across all three kingdoms of life. IscU serves as a scaffold for the assembly of intermediate iron-sulfur cluster centers and further mediates delivery to apo protein targets. Several proteins that mediate delivery of single metal ions to apo targets (termed metallochaperones) have recently been characterized structurally. Each displays a ferredoxin-like betaalphabetabetaalphabeta motif as a structural core. Assembly and delivery of a polynuclear iron-sulfur cluster is, however, a more complex pathway and presumably would demand a distinctive protein mediator. Here, we demonstrate Thermotoga maritima IscU (Tm IscU) to display unique structural and motional characteristics that distinguish it from other members of this class of proteins. In particular, IscU adopts a mobile, physiologically relevant, molten globule like state that is vastly different from the previously identified ferredoxin like fold that has thus far been characterized for other metallochaperones. The secondary structural content of Tm IscU is consistent with previous circular dichroism measurements on apo and holo protein, consisting of six alpha-helices and three beta-strands, the latter forming an anti-parallel beta-sheet. Extensive dynamics studies are consistent with a protein that has reasonably well defined secondary structural elements, but with a tertiary structure that is fluxional among widely different conformational arrangements. Analogous conformational flexibility does not exist in other structurally characterized metallochaperones; however, such a dynamic molecule may account for the lack of long-range NOEs, and allow both for the flexibility that is necessary for the multiple roles of Fe-S cluster assembly, and recognition and delivery of that cluster to a target protein. Additionally, the fluxionality of IscU is unique in that the protein appears to be more compact (based on 1H/2H exchange, R1, R2, and NOE data) but yet more fluid (lack of long-range NOEs) than typical molten globule proteins. PMID- 12909019 TI - Stereospecific interactions of proline residues in protein structures and complexes. AB - The constrained backbone torsion angle of a proline (Pro) residue has usually been invoked to explain its three-dimensional context in proteins. Here we show that specific interactions involving the pyrrolidine ring atoms also contribute to its location in a given secondary structure and its binding to another molecule. It is adept at participating in two rather non-conventional interactions, C-H...pi and C-H...O. The geometry of interaction between the pyrrolidine and aromatic rings, vis-a-vis the occurrence of the C-H...pi interactions has been elucidated. Some of the secondary structural elements stabilized by Pro-aromatic interactions are beta-turns, where a Pro can interact with an adjacent aromatic residue, and in antiparallel beta-sheet, where a Pro in an edge strand can interact with an aromatic residue in the adjacent strand at a non-hydrogen-bonded site. The C-H groups at the Calpha and Cdelta positions can form strong C-H...O interactions (as seen from the clustering of points) and such interactions involving a Pro residue at C' position relative to an alpha-helix can cap the hydrogen bond forming potentials of the free carbonyl groups at the helix C terminus. Functionally important Pro residues occurring at the binding site of a protein almost invariably engage aromatic residues (with one of them being held by C-H...pi interaction) from the partner molecule in the complex, and such aromatic residues are highly conserved during evolution. PMID- 12909020 TI - Transmembrane organization of the Bacillus subtilis chemoreceptor McpB deduced by cysteine disulfide crosslinking. AB - The Bacillus subtilis chemoreceptor McpB is a dimer of identical subunits containing two transmembrane (TM) segments (TM1, residues 17-34: TM2, residues 280-302) in each monomer with a 2-fold axis of symmetry. To study the organization of the TM domains, the wild-type receptor was mutated systematically at the membrane bilayer/extracytoplasmic interface with 15 single cysteine (Cys) substitutions in each of the two TM domains. Each single Cys substitution was capable of complementing a null allele in vivo, suggesting that no significant perturbation of the native tertiary or quaternary structure of the chemoreceptor was introduced by the mutations. On the basis of patterns of disulfide crosslinking between subunits of the dimeric receptor, an alpha-helical interface was identified between TM1 and TM1' (containing residues 32, 36, 39, and 43) and between TM2 and TM2' (containing residues 276, 277, 280, 283 and 286). Pairs of cysteine substitutions (positions 34/280 and 38/273) in TM1 and TM2 were used to further elucidate specific contacts within a monomer subunit, enabling a model to be constructed defining the organization of the TM domain. Crosslinking of residues that were 150-180 degrees removed from position 32 (positions 37, 41, and 44) suggested that the receptors may be organized as an array of trimers of dimers in vivo. All crosslinking was unaffected by deletion of cheB and cheR (loss of receptor demethylation/methylation enzymes) or by deletion of cheW and cheV (loss of proteins that couple receptors with the autophosphorylating kinase). These findings indicate that the organization of the transmembrane region and the stability of the quaternary complex of receptors are independent of covalent modifications of the cytoplasmic domain and conformations in the cytoplasmic domain induced by the coupling proteins. PMID- 12909021 TI - Structural analysis of lipid complexes of GM2-activator protein. AB - The GM2-activator protein (GM2-AP) is a small lysosomal lipid transfer protein essential for the hydrolytic conversion of ganglioside GM2 to GM3 by beta hexosaminidase A. The crystal structure of human apo-GM2-AP is known to consist of a novel beta-cup fold with a spacious hydrophobic interior. Here, we present two new structures of GM2-AP with bound lipids, showing two different lipid binding modes within the apolar pocket. The 1.9A structure with GM2 bound shows the position of the ceramide tail and significant conformational differences among the three molecular copies in the asymmetric unit. The tetrasaccharide head group is not visible and is presumed to be disordered. However, its general position could be established through modeling. The structure of a low-pH crystal, determined at 2.5A resolution, has a significantly enlarged hydrophobic channel that merges with the apolar pocket. Electron density inside the pocket and channel suggests the presence of a trapped phospholipid molecule. Structure alignments among the four crystallographically unique monomers provide information on the potential role for lipid binding of flexible chain segments at the rim of the cavity opening. Two discrete orientations of the S130-T133 loop define an open and a closed configuration of the hydrophobic channel that merges with the apolar pocket. We propose: (i) that the low-pH structure represents an active membrane-binding conformation; (ii) that the mobile S130-T133 loop serves as a gate for passage of ligand into the apolar pocket; and (iii) that this loop and the adjacent apolar V59-W63 loop form a surface patch with two exposed tryptophan residues that could interface with lipid bilayers. PMID- 12909022 TI - Adsorption isotherm and atomic force microscopy studies of the interactions between polymers and surfactants on steel surfaces in hydrocarbon media. AB - The adsorption isotherms for certain polymer and surfactant molecules (and in some cases their mixtures) on stainless steel beads from isooctane have been obtained, together with corresponding adsorbed layer thicknesses, using an atomic force microscope. The polymer is a terminally functionalised (ethylene diamine), low molecular weight polyisobutylene (PIB) derivative and the surfactants are basically alkyl or alkyl phenol alkoxylate molecules, which in one case has been derivatised with an amino functionality. The results indicate the presence of multilayers at the stainless steel-isooctane interface. Theoretical analysis of the surfactant adsorption isotherms suggests molecular aggregation at the interface with an aggregation number between 2 and 6, at the highest coverages. The adsorption of the polymer is reduced in the presence of the surfactant molecules. The polymer leaches metal ions from the steel surface at higher concentrations. PMID- 12909023 TI - New theoretical expressions for the five adsorption type isotherms classified by BET based on statistical physics treatment. AB - New theoretical expressions to model the five adsorption isotherm types have been established. Using the grand canonical ensemble in statistical physics, we give an analytical expression to each of five physical adsorption isotherm types classified by Brunauer, Emett, and Teller, often called BET isotherms. The establishment of these expressions is based on statistical physics and theoretical considerations. This method allowed estimation of all the mathematical parameters in the models. The physicochemical parameters intervening in the adsorption process that the models present could be deduced directly from the experimental adsorption isotherms by numerical simulation. We determine the adequate model for each type of isotherm, which fixes by direct numerical simulation the monolayer, multilayer, or condensation character. New equations are discussed and results obtained are verified for experimental data from the literature. The new theoretical expressions that we have proposed, based on statistical physics treatment, are rather powerful to better understand and interpret the various five physical adsorption type isotherms at a microscopic level. PMID- 12909024 TI - Vibrational study of the metal-adsorbate interaction of phenylacetic acid and alpha-phenylglycine on silver surfaces. AB - Raman and SERS spectra of phenylacetic acid and alpha-phenylglycine on silver sols have been recorded at several concentrations and pH values. The alpha phenylglycine has been also studied in D(2)O. The respective vibrational assignments have been proposed and the analysis of the SERS spectra has made it possible to conclude that phenylacetic acid links to the metal through its carboxylate group only, while alpha-phenylglycine links also through its amino group. In both cases the aromatic ring seems to be almost perpendicular to the metal surface. On the other hand, the contribution of the charge transfer (CT) mechanism to the enhancement of the SERS spectra has been calculated as well and it is found to be very important in both molecules. The band most enhanced by this mechanism is that of vibration 8a, mainly in alpha-phenylglycine. PMID- 12909025 TI - Kinetics and mechanisms of Zn complexation on metal oxides using EXAFS spectroscopy. AB - Zn(II) sorption onto Al and Si oxides was studied as a function of pH (5.1-7.52), sorption density, and ionic strength. This study was carried out to determine the role of the various reaction conditions and sorbent phases in Zn complexation at oxide surfaces. Extended X-ray absorption fine structure (EXAFS) spectroscopy was used to probe the Zn atomic environment at the metal oxide/aqueous interface. For both amorphous silica and high-surface-area gibbsite, Zn sorption kinetics were rapid and reached completion within 24 h. In contrast, Zn sorption on low-surface area-gibbsite was much slower, taking nearly 800 h for a sorption plateau to be reached. In the case of silica, EXAFS revealed that Zn was in octahedral coordination with first-shell oxygen atoms up to a surface loading of approximately 1 micro molm(-2), changing to tetrahedral coordination as surface loading and pH increased. For the high-surface-area gibbsite system, the Znz.sbnd;O first-shell distance was intermediate between values for tetrahedral and octahedral coordination over all loading levels. Zn formed inner-sphere adsorption complexes on both silica and high-surface-area gibbsite over all reaction conditions. For Zn sorption on low-surface-area gibbsite, formation of Znz.sbnd;Al layered double hydroxide (LDH) occurred and was the cause for the observed slow Zn sorption kinetics. The highest pH sample (7.51) in the Zn amorphous silica system resulted in the formation of an amorphous Zn(OH)(2) precipitate with tetrahedral coordination between Zn and O. Aging the reaction samples did not alter the Zn complex in any of the systems. The results of this study indicate the variability of Zn complexation at surfaces prevalent in soil and aquatic systems and the importance of combining macroscopic observations with methods capable of determining metal complex formation mechanisms. PMID- 12909026 TI - Visible-spectroscopy study of the adsorption of alizarinate by Al-montmorillonite in aqueous suspensions and in solid state. AB - The adsorption of the monovalent anionic dye alizarinate onto Na- and Al montmorillonite was carried out by adding the dye into aqueous clay suspensions. Electronic spectra of aqueous suspensions and of air-dried dye-clay complexes were studied. Na-montmorillonite adsorbed only part of the added dye. With total amount of alizarinate up to 5 mmol dye per 100 g clay the adsorption of the dye takes place on the broken bonds, leading to peptization of the clay. Al montmorillonite adsorbed alizarinate completely up to 10 mmol per 100 g clay. Above this loading there was a partition of the dye between the clay and the supernatant. The maximum adsorption for Na- and Al-clay was 4 and 25 mmol dye per 100 g clay, respectively. Absorption bands in the spectrum of Al-montmorillonite suspensions (488-504 nm) appear at longer wavelengths than in the spectrum of air dried Al-montmorillonite (415-455 nm). Thermo-X-ray study of these clay alizarinate complexes suggests that the organic compound was located in the interlayer space in Al-montmorillonite but was not located there in Na montmorillonite. In Al-montmorillonite alizarinate formed a coordination complex with exchangeable Al(3+). In Na-montmorillonite it formed bonds with Al exposed on the broken-bonds sites. PMID- 12909027 TI - PEG-catalytic water splitting in the interface of a bipolar membrane. AB - This paper investigates the effect of polyethylene glycol (PEG) on the water dissociation of bipolar membranes. To do this, bipolar membranes were prepared by immersing anion exchange membranes in different-concentration solutions of different-molecular-weight PEGs and then casting the solutions of sulfonated polyphenylene oxide (SPPO) on the anion exchange membranes. All the bipolar membranes with PEG in the interface are evaluated by current-voltage curves. The experimental results prove that PEG has excellent catalytic function for water dissociation. Furthermore, this function is enhanced by both PEG amount (PEG concentration) and PEG molecular weight in the interface of a bipolar membrane. PMID- 12909028 TI - Surface complexation modeling of uranium(VI) sorbed onto lanthanum monophosphate. AB - Sorption/desorption are basic processes in the field of contaminant transport. In order to develop mechanistically accurate thermodynamic sorption models, the simulation of retention data has to take into account molecular scale informations provided by structural investigations. In this way, the uranyl sorption constants onto lanthanum monophosphate (LaPO(4)) were determined on the basis of a previously published structural investigation. The surface complexation modeling of U(VI) retention onto LaPO(4) has been performed using the constant capacitance model included in the FITEQLv3.2 program. The electrical behavior of the solid surface was investigated using electrophoretic measurements and potentiometric titration experiments. The point of zero charge was found to be 3.5 and surface complexation modeling has made it possible to calculate the surface acidity constants. The fitting procedure was done with respect to the spectroscopic results, which have shown that LaPO(4) presents two kinds of reactive surface sites (lanthanum atoms and phosphate groups). The uranyl sorption edges were determined for two surface coverages: 40 and 20% of the surface sites that are occupied, assuming complete sorption. The modeling of these experimental data was realized by considering two uranyl species ("free" uranyl and uranyl nitrate complex) sorbed only onto phosphate surface groups according to the previously published structural investigation. The obtained sorption constants present similar values for both surface complexes and make it possible to fit both sorption edges: logK(U)=9.4 for z.tbnd;P(OH)(2)+UO(2)(2+)<- >z.tbnd;P(OH)(2)UO(2)(2+) and logK(UN)=9.7 for z.tbnd;P(OH)(2)+UO(2)NO(3)(+)<- >z.tbnd;P(OH)(2)UO(2)NO(3)(+). PMID- 12909029 TI - Thermal and FTIR spectroscopic analysis of the interactions of aniline adsorbed on to MCM-41 mesoporous material. AB - The adsorption of aniline on Na-AlMCM-41 synthesized by us has been characterized by infrared spectroscopy, temperature programmed desorption (TPD), and differential thermal analysis methods. Aniline would be mostly bound to the mesostructure through weak pi interactions. On the mesostructure containing adsorbed water, the co-adsorption of aniline could occur by weak hydrogen bonding through surface water molecules. For water, two possible modes of adsorption have been identified. Different associations between aniline and hydrated and nonhydrated mesostructures have been evaluated in order to favor the posterior in situ polymerization of adsorbed aniline. PMID- 12909030 TI - The adsorption of cationic and amphoteric copolymers on glass surfaces: zeta potential measurements, adsorption isotherm determination, and FT Raman characterization. AB - The adsorption of cationic and amphoteric copolymers onto controlled pore glass (CPG) powders has been studied by measurement of the powder particle zeta (zeta) potential, by determination of the adsorption isotherm, and by FT Raman measurements of the polymer-coated powder. The cationic polymers consisted chiefly of homopolymers of dimethyldiallylammonium chloride (DMDAAC) or copolymers of DMDAAC and acrylamide. The amphoteric polymers studied included copolymers of DMDAAC and acrylic acid. The comonomer ratio was varied to explore the dependence of cationic charge density on the extent and effect of adsorption. Both types of polymers adsorb onto the anionic glass surface via an ion-exchange mechanism. Consequently, a correspondingly higher mass of a low-charge-density copolymer adsorbs than of a cationic homopolymer. The presence of the anionic portion in the amphoteric polymers does not significantly alter this picture. The zeta potential, however, reflects the overall nature of the polymer. Cationic polymers effectively neutralize the glass surface, while amphoteric polymers leave the zeta potential net negative. Adsorption isotherms, determined via the depletion technique using colloidal titration, were used to "calibrate" a FT Raman method. The latter was used to determined the amount of adsorbed polymer under solution conditions in which colloidal titration could not be performed. PMID- 12909031 TI - A fractal analysis of analyte-estrogen receptor binding and dissociation kinetics using biosensors: environmental effects. AB - A fractal analysis is used to model the binding and dissociation kinetics between analytes in solution and estrogen receptors (ER) immobilized on a sensor chip of a surface plasmon resonance (SPR) biosensor. Both cases are analyzed: unliganded as well as liganded. The influence of different ligands is also analyzed. A better understanding of the kinetics provides physical insights into the interactions and suggests means by which appropriate interactions (to promote correct signaling) and inappropriate interactions such as with xenoestrogens (to minimize inappropriate signaling and signaling deleterious to health) may be better controlled. The fractal approach is applied to analyte-ER interaction data available in the literature. Numerical values obtained for the binding and the dissociation rate coefficients are linked to the degree of roughness or heterogeneity (fractal dimension, D(f)) present on the biosensor chip surface. In general, the binding and the dissociation rate coefficients are very sensitive to the degree of heterogeneity on the surface. For example, the binding rate coefficient, k, exhibits a 4.60 order of dependence on the fractal dimension, D(f), for the binding of unliganded and liganded VDR mixed with GST-RXR in solution to Spp-1 VDRE (1,25-dihydroxyvitamin D(3) receptor element) DNA immobilized on a sensor chip surface (Cheskis and Freedman, Biochemistry 35 (1996) 3300-3318). A single-fractal analysis is adequate in some cases. In others (that exhibit complexities in the binding or the dissociation curves) a dual fractal analysis is required to obtain a better fit. A predictive relationship is also presented for the ratio K(A)(=k/k(d)) as a function of the ratio of the fractal dimensions (D(f)/D(fd)). This has biomedical and environmental implications in that the dissociation and binding rate coefficients may be used to alleviate deleterious effects or enhance beneficial effects by selective modulation of the surface. The K(A) exhibits a 112-order dependence on the ratio of the fractal dimensions for the ligand effects on VDR-RXR interaction with specific DNA. PMID- 12909032 TI - Synthesis and physicochemical study of the laminin active sequence: SIKVAV. AB - The synthesis, physicochemical characterization, and interaction with membrane model systems of a peptide derived from the PA22-2 region of laminin are described. Surface activity studies indicate that this peptide is able to spread at the air-water interface being the maximal spreading pressure 20 mN/m at subphase concentrations around 10 micro M. Besides, these peptide molecules are also able to form stable monolayers. Physicochemical studies concerning the interaction of this peptide with lipids, organized in mono and bilayers, were carried out using Langmuir balance experiments and polarization fluorescence techniques. The peptide penetrates better in monolayers of DPPC than in those of PC and forms condensed mixed monolayers with DPPC. Energies of mixing are small thus indicating that deviations from ideality were almost negligible. Interactions with bilayers were studied through microviscosity changes (DPH and TMA-DPH probes), membrane permeability alterations (CF, NBD-PE/dithionite), and fusion promotion (NBD-PE/Rh-PE, resonance energy transfer). Results indicate that this sequence interacts very softly with bilayers without promoting changes in their organization. These data as well as the lack of interaction with erythrocytes suggest that coating liposomes with this peptide through chemical amide bonds can render stable inmunoliposomes for further biological applications. PMID- 12909033 TI - Localized changes in the structural stability of myoglobin upon adsorption onto silica particles, as studied with hydrogen/deuterium exchange mass spectrometry. AB - A new method is presented for monitoring the conformational stability of various parts of a protein that is physically adsorbed onto nanometer-sized silica particles. The method employs hydrogen/deuterium (H/D) exchange of amide hydrogens, a process that is extremely sensitive to structural features of proteins. The resulting mass increase is analyzed with Fourier transform ion cyclotron resonance (FTICR) mass spectrometry. Higher structural specificity is obtained by enzymatically cleaving the adsorbed proteins prior to mass spectrometric analysis. The mass increases of four peptic fragments of myoglobin are followed as a function of the H/D exchange time. The four peptic fragments cover 90% of the myoglobin structure. Two of the peptic fragments, located in the middle of the myoglobin sequence and close to the heme group, do not show any adsorption-induced changes in their structural stability, whereas the more stable C- and N-terminal fragments are destabilized. Interestingly, for the N-terminal fragment, comprising residues 1-29, two distinct and equally large conformational populations are observed. One of these populations has a stability similar to that in solution (-23 kJ/mol), whereas the other population is highly destabilized upon adsorption (-11 kJ/mol). PMID- 12909034 TI - Gold nanolayer-encapsulated silica particles synthesized by surface seeding and shell growing method: near infrared responsive materials. AB - Gold nanolayer-encapsulated silica particles whose optical resonance is located in 750-900 nm spectral region were synthesized by combining Sn (tin)-surface seeding and a shell growing process. The synthesized composite particles can be potentially used in wide biological fields, due to biocompatibility and a well known bioconjugation technique of gold layer. Sn atoms, which can act not only as a catalytic surface for reduction of gold but also as a linker between silica surface and gold nanoparticles, were chemically deposited on hydroxylated silica particles. Then, we introduced another reductant with gold chloride in order to produce a multilayer of Au shell. In the process, Au shells grew by the reduction of additional gold ions on the Sn-functionalized silica surface and resulted in the subsequent coalescence and growth of the deposited gold nanoparticles. Finally, a complete gold nanoshell was formed on the silica surface by the one step method, without a repeated coating process. The deposition of a gold nanolayer on the silica particles was easily controlled by the concentration ratio of Sn-functionalized silica particles and gold chloride solutions. Transmission electron microscopy (TEM) images and optical extinction spectra clearly showed that gold nanolayers were successfully deposited on the silica surface by the novel method. As the gold colloids attached on the silica surface grew, their optical plasmon peak became red-shifted until complete a gold shell was formed. After the gold shell was completed, the optical plasmon resonance became blue-shifted and the extinction spectra were functions of a relative ratio of the core to shell thickness. PMID- 12909035 TI - Influence of solvent on the growth of ZnO nanoparticles. AB - We have synthesized ZnO nanoparticles by precipitation from zinc acetate in a series of n-alkanols from ethanol to 1-hexanol as a function of temperature. In this system, nucleation and growth are relatively fast and, at longer times, the average particle size continues to increase due to diffusion-limited coarsening. During coarsening, the particle volume increases linearly with time, in agreement with the Lifshitz-Slyozov-Wagner (LSW) model. The coarsening rate increases with increasing temperature for all solvents and increases with alkanol chain length. We show that the rate constant for coarsening is determined by the solvent viscosity, surface energy, and the bulk solubility of ZnO in the solvent. PMID- 12909036 TI - Unexpected effects of trace impurities on the properties of polymer-stabilized ruthenium colloids from different sources of ruthenium(III) chloride hydrate. AB - The stability and the catalytic performances of polyvinylpyrrolidone-stabilized ruthenium colloids (PVP-Ru) prepared by using RuCl(3).xH(2)O of different sources have been investigated. The trace impurity content of platinum and palladium in RuCl(3).xH(2)O was demonstrated to be the main factors affecting the properties of PVP-Ru colloids. PMID- 12909037 TI - Silica nanobottles templated from functional polymer spheres. AB - Nanosized hollow silica spheres with holes in the wall (denoted as silica nanobottles) have been successfully prepared by assembly of functional polymer nanospheres with tetraethoxysilane (TEOS) through hydrothermal methods, coupled with removal of the core by programmed calcination. The functional polymer nanospheres were obtained by emulsifier-free emulsion copolymerization of styrene and (ar-vinylbenzyl) trimethylammoium chloride. The silica nanobottle sample was characterized by thermogravimetric analysis (TG), differential thermal analysis (DTA), transmission electron microscopy (TEM), and nitrogen adsorption techniques. The above characterizations confirm that the silica nanobottles have holes of about 8 nm in the wall and this unique structural feature might be useful for their encapsulation. Furthermore, characterization by scanning electron microscopy (SEM), energy dispersive X-ray analysis (EDX), and UV-visible absorption (UV-vis) showed that the luminescent material Eu(TTA)(3)(TPPO)(2) could be effectively encapsulated in silica nanobottles. This reveals that silica nanobottles have potential applications for nanotechniques. PMID- 12909038 TI - The Christiansen effect of brightly colored colloidal dispersion with an amphiphilic polymer. AB - A novel coloration phenomenon in a colloidal dispersion with an amphiphilic polymer was found. The dispersion consists of tetrahydrofuran (THF), an aqueous solution of sodium thiosulfate (Na(2)S(2)O(3).5H(2)O), and hydroxypropylcellulose (HPC). The dispersion was emulsified by HPC as an amphiphilic polymer, so that the aqueous phase was confined in droplets in the THF matrix. It typically appeared bluish violet at room temperature and turned into blue with increasing temperature. In this system, the refractive indices of the inside and outside of the droplet coincided at a certain wavelength at which the light passes through without scattering, which is called the Christiansen effect. The color observed was successfully simulated by Mie's scattering theory in combination with the Christiansen effect. PMID- 12909039 TI - Polarizability and complex conductivity of dilute suspensions of spherical colloidal particles with charged (polyelectrolyte) coatings. AB - The dielectric relaxation of polyelectrolyte-coated colloidal particles is examined via "exact" numerical solutions of the governing electrokinetic equations. The charged polymer coatings are characterized by a nominal charge density, thickness, and permeability. Brush-like segment density profiles are considered here, but more sophisticated segment and charge density profiles are accommodated by the model. The role of added counterions and nonspecific adsorption is considered briefly before examining how the experimentally measured conductivity and dielectric constant increments reflect the frequency of the applied electric field, the strength of the electrolyte, and characteristics of the polymer coatings, namely the charge, charge density, and permeability. Finally, a strategy is suggested by which dielectric spectroscopy and electrophoresis can be used to characterize polymer-coated particles. This approach complements experiments where electroviscous effects such as dynamic light scattering and sedimentation are weak. PMID- 12909040 TI - Hydrodynamic drag force exerted on activated sludge floc at intermediate Reynolds number. AB - We hung the activated sludge flocs on an elastic nylon stick and then subjected it to a uniform water flow and measured its displacement. The hydrodynamic drag force exerted on the floc was subsequently estimated, both for cationic flocculated flocs and for flocculated and then frozen/thawed flocs. A confocal laser scanning microscope (CLSM) was employed to probe the interior structure of flocs. Polyelectrolyte flocculation leads to a compact global structure, and hence high drag force exerted on the floc by water. The corresponding C(D)Omega value at Re=12-27 for flocs ranges from 1.58 to 3.61. Fast freezing would little affect the hydrodynamic drag force. Slow freezing, in contrast, considerably consolidated the floc structure and hence presented impermeable sphere-like behavior of the slowly frozen/thawed flocs. PMID- 12909041 TI - Role of interaction forces in controlling the stability and polishing performance of CMP slurries. AB - Chemical mechanical polishing (CMP) is an essential step in metal and dielectric planarization in multilayer microelectronic device fabrication. In the CMP process it is necessary to minimize the extent of surface defect formation while maintaining good planarity and optimal material removal rates. These requirements are met through the control of chemical and mechanical interactions during the polishing process by engineering the slurry chemistry, particulate properties, and stability. In this study, the performance of surfactant-stabilized silica CMP slurries at high pH and high ionic strengths are investigated with particular emphasis on the particle-particle and particle-substrate interactions. It is shown that for the design of consistently high performing slurries, stability of abrasive particles must be achieved under the dynamic processing conditions of CMP while maintaining sufficient pad-particle-wafer interactions. PMID- 12909042 TI - Reductive desorption of 11-mercaptoundecanoic acid monolayers modified by covalent attachment of 1,3- and 1,4-phenylenediamine. AB - 1,3- and 1,4-phenylenediamine were covalently attached to a self-assembled monolayer of 11-mercaptoundecanoic acid by formation of an amide bond. Bonding of the phenylenediamine isomers was confirmed by the presence of infrared features attributable to ring vibrational modes, the amide I vibration, and the amide II vibration in the reflection-absorption infrared spectrum of the modified interface. Reductive desorption peak potentials were shifted by 120 mV for the monolayer modified by 1,4-phenylenediamine and by 90 mV for the monolayer modified by 1,3-phenylenediamine relative to the desorption potential of the unmodified 11-mercaptoundecanoic acid monolayer. These potential shifts represent 23 and 17 kJ/mol stabilization of the monolayers modified by 1,4- and 1,3 phenylenediamine, respectively. Some of the additional stabilization energy is attributed to lateral hydrogen bonding between the amide groups and unreacted carboxylic acid groups in the monolayer. The difference between the stabilization energies for the interfaces modified with the different phenylenediamine isomers is attributed to structural differences between the isomeric monolayers influencing the molecular packing of the modified interface. PMID- 12909043 TI - Ion-specific effect on the gelation kinetics of concentrated colloidal silica suspensions. AB - The comparative gelation kinetics of concentrated colloidal silica suspensions at pH 9.8 has been studied in the presence of two different counterions, ammonium as a representative structure-breaker ion and sodium as a representative structure maker ion. For the same ionic strength of 0.45 M, the experimental results have indicated a pronounced effect of the ion-specificity on the aggregation kinetics and on the microstructure of the gelified networks at the sol-gel transition. This was ascribed to the significant difference of the critical coagulation concentration value in the presence of the two electrolytes. PMID- 12909044 TI - Adsorption and isotope effects by ion exchange with 1-aza-12-crown-4 bonded Merrifield peptide resin. AB - Magnesium isotope effects were investigated by chemical ion exchange with synthesized 1-aza-12-crown-4 bonded Merrifield peptide resin using elution chromatography. The capacity of azacrown ion exchanger was 0.89 meq/g dry resin. The heavier isotopes of magnesium were enriched in the resin phase, while the lighter isotopes were enriched in the solution phase. The hydration effect is less than the complexation and isotope mass effects. The single stage separation factor was determined according to the method of Glueckauf from the elution curve and isotopic assays. The separation factors of 24Mg(2+)-25Mg(2+), 24Mg(2+) 26Mg(2+), and 25Mg(2+)-26Mg(2+) were 1.012, 1.023, and 1.011, respectively. PMID- 12909045 TI - Structural and adsorptive properties of activated carbons prepared by carbonization and activation of resins. AB - Four activated carbons (S1-S4) possessing different structural characteristics were prepared by carbonization of commercial resins (used for ion exchange) and subsequent activation. Their textural parameters were determined on the basis of nitrogen adsorption-desorption at 77.4 K, analyzed by applying several local and overall adsorption isotherm equations. The nature of carbon surface functionalities was analyzed by FTIR spectroscopy. The GC and solid-phase extraction (SPE) techniques were applied to study the influence of the texture of carbonaceous materials on their adsorptive properties. The adsorption efficiency of synthesized carbons with respect to alkylhalides used as probe compounds in the GC measurements varied over a range from 28% (C(2)H(3)Cl(3)/S2) to 85% (CHBr(3)/S1) depending on the type of adsorbates and adsorbents. The concentrating efficiency of these carbons in SPE of explosive materials changed over a larger range from 12% (trinitroglycerin/S4) and 13% (trinitrotoluene/S2) up to 100% (octogen/S1). Active carbon prepared using Zerolite 225x8 as a precursor demonstrated better results than other carbons in two types of adsorption with average values of the efficiency of 75.4% for explosives and 60.8% for alkylhalides. PMID- 12909046 TI - An alternative Avrami equation to evaluate kinetic parameters of the interaction of Hg(II) with thin chitosan membranes. AB - In this work, thin chitosan membranes were utilized as an adsorbent for the removal of Hg(II) from aqueous solutions. A rise of temperature accelerates the mass transfer of Hg(II) to the membranes, surfaces. The kinetic data did not present a good fitting to the traditional Lagergren adsorption kinetic equations. An alternative Avrami kinetic equation was employed and successfully fitted to the kinetic adsorption quantities. From this new equation, two regions presenting distinct kinectic parameters were found, and the use of the parameter n was also relationed to the determination of the kinetic order. Variations of the adsorption kinetic rate in relation to the time, the initial Hg(II) concentration, and the temperature were also calculated and are discussed. PMID- 12909047 TI - Cementation reactions in the presence of nitrogen compounds. AB - The effect of aniline derivatives on the rate of cementation of copper from dilute copper sulfate solution on iron rods was investigated. The variables studied were concentration of CuSO(4), temperature, organic additives, and their concentrations. It was found that the cementation reaction was first-order in the presence and in the absence of additives. Nitrogen compounds were found to decrease the rate of cementation by 10 to 80% depending on the type of inhibitor as well as its concentration.The adsorption isotherm, the effect of temperature, and the percentage inhibition show that the order of inhibition is p chloroaniline>o-chloroaniline>m-chloroaniline and p-chloroaniline>p-aminobenzoic acid>p-aminoacetophenone. The thermodynamic parameters DeltaS( not equal ), DeltaG( not equal ), DeltaH( not equal ) were given. PMID- 12909048 TI - Calculation of surface enthalpy of solids from an ab initio electronegativity based model: case of ice. AB - A very simple route to calculation of the surface energy of solids is proposed because this value is very difficult to determine experimentally. The first step is the calculation of the attractive part of the electrostatic energy of crystals. The partial charges used in this calculation are obtained by using electronegativity equalization and scales of electronegativity and hardness deduced from physical characteristics of the atom. The lattice energies of the infinite crystal and of semi-infinite layers are then compared. The difference is related to the energy of cohesion and then to the surface energy. Very good results are obtained with ice, if one compares with the surface energy of liquid water, which is generally considered a good approximation of the surface energy of ice. PMID- 12909049 TI - On air entrainment in coatings. AB - A series of experiments that clarify how air bubbles become entrained into coatings are described. The contact line dynamics at the air-liquid interface surrounding a fiber is characterized for a typical coating die operating under atmospheric and pressurized conditions. Glycerin and other viscous liquids are used to reveal that a critical fiber speed exists at which air entrainment begins. The observations confirm that the critical capillary number Ca(c) depends on the physical properties of the coating material, in the form of the Morton number. When the liquid supply is pressurized, the experiments show that adjusting the pressure can stabilize the displaced free surface interface at a prescribed location. Controlling the meniscus location in this way eliminates air entrainment. The threshold occurs when the applied pressure balances the shear exerted on the coating by the moving fiber. Using this approach it is possible to eliminate air entrainment and attain stable wetting at very large values of the capillary number, e.g., Ca congruent with 50. PMID- 12909050 TI - Wetting phase connectivity and irreducible saturation in simple granular media. AB - We present drainage simulations that allow trapping of wetting phase in a simple but nontrivial granular medium, a dense random packing of equal spheres. The basis for the simulations is a network model derived directly from the known locations and dimensions of pore space features. This provides a means of evaluating the morphology of trapped wetting phase. The possible morphologies depend on the assumed connectivity of the wetting phase. At one extreme, we assume that the entire wetting phase except for pendular rings is connected. At the opposite extreme, we illustrate a low level of connectivity by assuming that pendular rings are trapped as soon as the pores surrounding them are drained; any wetting phase not yet drained from pore throats connecting these pores is also assumed to be trapped. Finally we consider a set of criteria involving larger neighborhoods within the network, which allows trapping in individual pores. Irreducible wetting phase saturations obtained in the latter case agree with experimental data. The numbers of pendular rings and liquid bridges are also consistent with observations. Because the agreement does not involve adjustable parameters, we conclude that a relatively simple, local evaluation of trapping criteria can yield physically representative wetting phase configurations. PMID- 12909051 TI - Wetting behavior of flax fibers as reinforcement for polypropylene. AB - The wetting behavior of several flax (cellulose as reference) and polypropylene fibers is characterized by measuring the wetting rates (penetration velocities) of a series of liquids using the capillary rise technique. This present paper aims to provide a deeper understanding of the complex nature of natural fibers and their surface properties. The fiber surface tensions are estimated from plots of the normalized wetting rate as a function of the surface tension of the liquids assuming, in analogy to Zisman's method, that the maximum of the normalized wetting rate corresponds to the solid surface tension. The estimated surface tensions of the investigated flax fibers indicate that all the fibers are quite "hydrophobic." The method used to separate the fibers from the rest of the plants has a large influence on the estimated fiber surface tensions. In the case of polypropylene (PP) fibers, the estimated surface tension corresponds well with literature data. Grafting small amounts of maleic acid anhydrite (MAH) onto the PP surfaces will not affect the wetting behavior and, therefore, the surface tension, whereas grafting larger amounts (10 wt%) of MAH causes the polymer surface tension to increase significantly. Additional pH-dependent zeta-potential measurements show that even the "pure" PP-fibers contain acidic surface functions, possibly due to further processing at elevated temperatures (thermal degradation or other aging processes). PMID- 12909052 TI - Characterization of a quaternary liquid system improving the bioavailability of poorly water soluble drugs. AB - The aim of this work is the characterization of the quaternary system composed of water, triacetin (oil), ethanol (alcohol), and Tween 80 (surfactant), as its results enable the enhancement of the bioavailability of nimesulide, a poorly water soluble nonsteroidal antiinflammatory drug widely employed in the pharmaceutical field. Particular attention is devoted to the surfactant-free ternary system, as it proved able to solubilize nimesulide as well, and the absence of a surfactant is desirable in order to keep the preparation as tolerable as possible. Both bulk and interfacial properties of this system are investigated, and a mathematical model to calculate the interface composition of a three-component two-phase system is developed. This model is based on Gibbs' theory on interfaces, which considers an arbitrary mathematical dividing surface so that the two phases continue uniformly up to it, although interface regions have no sharply defined boundaries. We find that both the quaternary and the ternary systems investigated show a miscibility lacuna and that, in the surfactant-free ternary system, an increase of the ethanol weight fraction is reflected as an impoverishment of the ethanol interfacial molar fraction. PMID- 12909053 TI - Micellization and catalytic activity of the cetyltrimethylammonium bromide-Brij 97-water mixed micellar system. AB - Surface tension measurements and the kinetic study of the basic hydrolysis of ethyl p-nitrophenyl chloromethyl phosphonate were used to examine the structural behavior and catalytic activity of the cethyltrimethylammonium bromide (CTAB) polyoxyethylene (10) oleyl ether, C(18)H(35)(OCH(2)CH(2))(10)OH (Brij 97)-water mixed micellar system. Application of the regular solution model to the experimental data yields the value of the interaction parameter beta as -4.6, which indicates an attractive interaction of the surfactants in the mixed micelle and reflects synergistic solution behavior of the mixture. The mixed micellar composition is found to be enriched in the surfactant with the lower critical micelle concentration (cmc). In the kinetic study a nonmonotonic change in the pseudo-first-order rate constant of basic hydrolysis of the substrate is observed with increasing mole fraction of nonionic surfactant. The pseudophase micellar model reveals that the concentration factor mainly contributes to the catalytic effect, while the microenvironmental factor plays a negative role. PMID- 12909054 TI - New thermodynamic/electrostatic models of adsorption and tension equilibria of aqueous ionic surfactant mixtures: application to sodium dodecyl sulfate/sodium dodecyl sulfonate systems. AB - The nonideal adsorbed solution (NAS) theory has been formally extended to adsorption at the air/water interface from aqueous mixtures of ionic surfactants, explicitly accounting for the surface potential of the adsorbed monolayer with the Gouy-Chapman theory. This new ionic NAS (iNAS) theory is thermodynamically consistent and, when coupled to a micellization model, is valid for concentrations below and above the mixed cmc. Counterion binding is incorporated into the model using two fractional binding parameters, beta(sigma) for the adsorbed monolayer and beta(m) for the micelles. The regular solution theory is used to model the nonideal interactions within the adsorbed monolayer and within the mixed micelles. New tension data for an equimolar mixture of sodium dodecyl sulfate (SDS) and sodium dodecyl sulfonate (SDSn) at two salinities fit this model well when mixing is ideal. The total surface densities, the surface compositions, and the surface potentials for the mixed monolayers are calculated. When there is no added salt, at total surfactant concentrations below the mixed cmc, the adsorbed monolayer is enriched in SDSn, but at total concentrations at and above the mixed cmc, the adsorbed monolayer is nearly an equimolar mixture. In the presence of 100 mM NaCl, the adsorbed monolayer is nearly an equimolar mixture, independent of the total surfactant concentration. PMID- 12909055 TI - Fluorescent behavior of B-phycoerythrin in microemulsions of aerosol OT/water/isooctane. AB - Taking advantage of its unusual fluorescent properties, the incorporation of B phycoerythrin (B-PE) in aerosol OT (AOT, sodium bis-(2-ethylhexyl) sulphosuccinate)/water/isooctane microemulsions was investigated by following their steady-state and time-resolved fluorescence as a function of the water-to surfactant molar ratio, w(0). The fluorescent intensity at 575 nm increased continuously with increasing water content, saturating at a w(0) around 35 and staying practically constant at w(0)> or =40. The steady-state anisotropy showed an initial increase with increasing water content until w(0)=23 and then decreased strongly, staying practically constant when w(0)> or =40. The values of the fluorescent parameters, anisotropy and fluorescent intensity, were unchanged when the water content of the system increased in the range between w(0)=40 to 50. This implies the effective incorporation of B-PE in the microemulsion droplets with w(0)> or =40, as well as the equilibrium of the dispersion at these water/surfactant ratios, since higher water content does not affect the main surrounding microenvironment of the protein. The overall incorporation in the microemulsion droplets caused minor spectroscopic changes with respect to biliprotein in aqueous solution of 20 mM sodium phosphate buffer, pH 7.0, such as a blue absorption shift of 3 nm and an emission shift of 1.5 nm, as well as a slight increase in excitation anisotropy spectrum mainly caused by a decrease in protein mobility. Therefore, there are no important interactions between the chromophores and the AOT sulfonate head groups. Emission intensity decays followed complex kinetics in both aqueous and dispersion media. The stability with time and temperature of the biliprotein in the microemulsion was higher than in the aqueous solution. All the results can be explained in terms of B-PE inclusion in the water droplets of AOT microemulsions where the protein has similar configuration and conformation to that in aqueous solution but with the chromophores more protected. PMID- 12909056 TI - Equilibrium partition of polycyclic aromatic hydrocarbons in a cloud-point extraction process. AB - A cloud-point extraction (CPE) process using the nonionic surfactant Tergitol 15 S-7, a secondary ethoxylated alcohol, to extract selected polycyclic aromatic hydrocarbons (PAHs) from aqueous solutions is investigated. The CPE process is facilitated at the ambient temperature, ca. 22 degrees C, by the reduction of the cloud-point temperature of the surfactant solution by addition of sodium sulfate. It is observed that the preconcentration factor could be enhanced either by increasing the salt concentration or by decreasing the initial surfactant concentration in the micellar solution. A high preconcentration factor of about 40 was achieved at 1 wt% surfactant concentration with the addition of 0.6 M Na(2)SO(4). It is also noted that the equilibrium partition coefficients of the model PAHs are nearly independent of surfactant concentrations, up to 3 wt%, in this study. Correlations between the equilibrium partition coefficients K(p) of the PAHs and their octanol-water partition coefficients K(ow), as well as K(p) and the molar volume V(x) of these PAHs, indicate that the partition processes of the PAHs in the CPE processes are mainly governed by their hydrophobic affinities to the surfactant aggregates. Furthermore, the effect of added Na(2)SO(4) on the equilibrium partition coefficients is also studied. It is shown that addition of more Na(2)SO(4) to the surfactant solution gives more partition of the PAHs into the surfactant-rich phase. PMID- 12909057 TI - Mechanism for defoaming by oils and calcium soap in aqueous systems. AB - The effect of oils, hardness, and calcium soap on foam stability of aqueous solutions of commercial surfactants was investigated. For conditions where negligible calcium soap was formed, stability of foams made with 0.1 wt% solutions of a seven-EO alcohol ethoxylate containing dispersed drops of n hexadecane, triolein, or mixtures of these oils with small amounts of oleic acid could be understood in terms of entry, spreading, and bridging coefficients, i.e., ESB analysis. However, foams made from solutions containing 0.01 wt% of three-EO alcohol ethoxysulfate sodium salt and the same dispersed oils were frequently more stable than expected based on ESB analysis, reflecting that repulsion due to overlap of electrical double layers in the asymmetric oil-water air film made oil entry into the air-water interface more difficult than the theory predicts. When calcium soap was formed in situ by the reaction of fatty acids in the oil with calcium, solid soap particles were observed at the surfaces of the oil drops. The combination of oil and calcium soap produced a synergistic effect facilitating the well-known bridging instability of foam films or Plateau borders and producing a substantial defoaming effect. A possible mechanism of instability involving increases in disjoining pressure at locations where small soap particles approach the air-water interface is discussed. For both surfactants with the triolein-oleic acid mixtures, calculated entry and bridging coefficients for conditions when calcium soap formed were positive shortly after foam generation but negative at equilibrium. These results are consistent with the experimental observation that most defoaming action occurred shortly after foam generation rather than at later times. PMID- 12909058 TI - Electrokinetic flow in a capillary with a charge-regulating surface polymer layer. AB - An analytical study of the steady electrokinetic flow in a long uniform capillary tube or slit is presented. The inside wall of the capillary is covered by a layer of adsorbed or covalently bound charge-regulating polymer in equilibrium with the ambient electrolyte solution. In this solvent-permeable and ion-penetrable surface polyelectrolyte layer, ionogenic functional groups and frictional segments are assumed to distribute at uniform densities. The electrical potential and space charge density distributions in the cross section of the capillary are obtained by solving the linearized Poisson-Boltzmann equation. The fluid velocity profile due to the application of an electric field and a pressure gradient through the capillary is obtained from the analytical solution of a modified Navier-Stokes/Brinkman equation. Explicit formulas for the electroosmotic velocity, the average fluid velocity and electric current density on the cross section, and the streaming potential in the capillary are also derived. The results demonstrate that the direction of the electroosmotic flow and the magnitudes of the fluid velocity and electric current density are dominated by the fixed charge density inside the surface polymer layer, which is determined by the regulation characteristics such as the dissociation equilibrium constants of the ionogenic functional groups in the surface layer and the concentration of the potential-determining ions in the bulk solution. PMID- 12909059 TI - Viscosity measurements of a ferrofluid: comparison with various hydrodynamic equations. AB - Effective viscosity of a magnetic fluid as a function of applied magnetic field oriented in the perpendicular direction of the capillary flow is determined. Close agreement with the Shliomis expression derived on the basis of effective field method is observed. PMID- 12909060 TI - Covalently attached sandwich structure from colloidal particles and diazoresin. AB - A covalently attached sandwich structure between layers and particles has been fabricated from z.sbnd;COOH-containing copolymer latex particles and z.sbnd;N(2)(+)-containing polymers by self-assembly combined with a UV irradiation technique. The ionic bonds involving the layers and particles change to covalent bonds under UV irradiation and the sandwich structure become very stable toward polar organic solvents and electrolyte aqueous solutions. PMID- 12909061 TI - Nanosecond time-resolved studies of long-lived photoinduced charge separation in the dyad fluorescein-anthraquinone-methyl ester adsorbed on TiO2 colloids. AB - A dyad composed of fluorescein and 2-methyl-anthraquinone (FL-AQ) was synthesized and its photophysical properties were examined by absorption, fluorescence spectroscopy, and fluorescence lifetime. The charge-separated state formed by photoinduced intramolecular electron transfer was detected by nanosecond transient absorption spectroscopy for the first time. When FL is excited in solution, the photoinduced electron transfer from FL to AQ proceeds efficiently. The rate constant and the efficiency of intramolecular electron transfer are 3.95 x 10(9) s(-1) and 95%, respectively. Its charge-separated state lifetime is too short to detect by transient absorption spectroscopy. Adding nanometer colloidal TiO(2) to an FL-AQ ethanol solution prolongs the lifetime of the charge-separated state, so its transient absorption signal is recorded significantly. The lifetimes of FL(+). at 480 nm and AQ(-). at 560 nm in the FL-AQ/TiO(2) colloidal system are 11.1 and 8.93 mivros, respectively. PMID- 12909062 TI - Void closure and interdiffusion processes during latex film formation from surfactant-free polystyrene particles: a fluorescence study. AB - This study reports a steady state fluorescence (SSF) technique for studying film formation from surfactant-free polystyrene (PS) latex particles. The latex films were prepared from pyrene (P)-labeled PS particles at room temperature and annealed at elevated temperatures for 5-, 10-, 20-, and 30-min time intervals above the glass transition (T(g)) temperature of polystyrene. During the annealing processes, the transparency of the film changed considerably. Scattered light (I(sc)) and fluorescence intensity (I(0P)) from P were measured after each annealing step to monitor the stages of film formation. Evolution of transparency of latex films were monitored using photon transmission intensity, I(tr). Scanning electron microscopy (SEM) was used to detect variation in the physical structure of annealed films. Onset temperature for film formation, T(0), void closure, T(v), and healing temperatures, T(h), were determined and corresponding activation energies were measured. Void closure and interdiffusion stages were modeled and related activation energies were determined. PMID- 12909063 TI - The interaction energy between two parallel plates with constant surface charge density. AB - On the basis of Langmuir's suggestion we simplify the Poisson-Boltzmann equation and derive the relation of surface potential, potential midway, and the plate distance. Thus we obtain the interaction force and energy equations between two dissimilar plates in the case of constant surface charge density. Agreement with the exact numerical values of the interaction of dissimilar plates is good. This method may not only apply to the cases of high constant potential but to the case of high constant charge density. PMID- 12909064 TI - Anilinepropylsilica xerogel used as a selective Cu (II) adsorbent in aqueous solution. AB - The metal ion adsorption properties of the microporous hybrid anilinepropylsilica xerogel were studied using divalent copper, zinc, and cadmium ions in aqueous solutions in concentrations ranging from 10(-4) up to 5x10(-3) moll(-1). At low concentrations the surface of the solid phase presents selectivity for Cu (II), even in competitive conditions. This preferential sorption ability for copper in relation to zinc and cadmium ions was interpreted by considering the xerogel morphology. PMID- 12909065 TI - Early invasive therapy of non ST-elevation acute coronary syndromes--combined with upstream antiplatelet therapy: yes--but how early? PMID- 12909066 TI - Diabetes abolishes the gender gap in coronary heart disease. PMID- 12909067 TI - Quality of life in nonpharmacologic treatment of atrial fibrillation. AB - The mainstay of treatment for atrial fibrillation, AF, remains pharmacologic control, either by maintaining sinus rhythm or by controlling the ventricular rate and allowing AF to continue. In patients where pharmacologic therapy i not effective, not tolerated or contraindicated, nonpharmacologic treatment may be beneficial. In the last two decades the number of nonpharmacologic treatment options (catheter ablation, cardiac pacing, internal defibrillation, and dysrhythmia surgery) for AF have markedly increased and the number of patients undergoing such treatment is steadily increasing. The most important reason for these treatment strategies is the hope of reducing symptoms, preventing complications and improving quality of life, QoL. However, the impact of nonpharmacologic therapy on QoL is far from established. Following a short presentation of the basic definitions and instruments used in QoL research the present paper reviews clinical studies that have assessed QoL in patients undergoing nonpharmacologic treatment of AF. Major limitations and methodological problems are emphasized. Among these are highly selected often-heterogeneous patients groups, small size, lack of control group and the use of non-validated QoL instruments. Furthermore, in most studies antiarrhythmic medication have been discontinued at the time of the intervention and it is not clear to which degree the improvement in QoL is related solely to the nonpharmacologic treatment or to the removal of drug related adverse effects. Although the currently available data from adequately designed studies are sparse and further investigations are needed, it is noteworthy that the majority of patients undergoing nonpharmacologic treatment report enhanced QoL. PMID- 12909068 TI - A comparison of two invasive strategies in patients with non-ST elevation acute coronary syndromes: results of the Early or Late Intervention in unStable Angina (ELISA) pilot study. 2b/3a upstream therapy and acute coronary syndromes. AB - BACKGROUND: Only few studies specifically addressed the effect of timing of angiography and/or pre-treatment with a glycoprotein 2b/3a receptor blocker in patients with non-ST elevation acute coronary syndromes (ACS) who undergo invasive treatment. METHODS: In a 2-year period, 220 patients with non-ST elevation ACS, were randomized to early angiography without tirofiban pre treatment (Early strategy) or to delayed angiography after 24-48h pre-treatment with tirofiban (Late strategy). The first 48h after admission, CKmb levels were measured and enzymatic infarct size (LDHQ(48)) was assessed by the area under the LDH release curve. When PCI was performed beyond 48h, CKmb was measured 6 and 12h afterwards. RESULTS: Median time to angiography was 6 (Early) and 50 (Late) hours. PCI was performed in 130 patients (59%). In these patients, a patent (TIMI 2 or 3 flow) culprit vessel was more often present in the Late group compared to the Early group (66% vs 82% p=0.05). In patients with an elevated CKmb (n=96, 44%), LDHQ(48)was significantly lower in patients who underwent angiography after pre-treatment with tirofiban (629+/-503U/L (Early) vs 432+/-441U/L (Late), p=0.02). No difference in clinical outcome between the groups was observed at 30 days follow-up. CONCLUSION: This pilot study showed that a strategy of delayed angiography with concomitant pre-treatment with tirofiban is associated with improved angiographic outcomes and less initial enzyme release, compared to a strategy of immediate angiography without 2b/3a inhibitor pre-treatment. The use of an end point parameter, which assess total enzyme release over a given period of time, might be of special value in patients with non-ST elevation ACS, who undergo very early invasive treatment. PMID- 12909069 TI - Cardiovascular events in type 2 diabetes: comparison with nondiabetic individuals without and with prior cardiovascular disease. 10-year follow-up of the Hoorn Study. AB - AIMS: We questioned whether prior cardiovascular disease has the same impact on risk of cardiovascular events as type 2 diabetes, and whether this differed between men and women. METHODS AND RESULTS: To address these issues we compared the 10-year risk of cardiovascular events among 208 Caucasian individuals with diabetes to that of 2253 Caucasian individuals without diabetes, in a population based cohort study. Gender significantly modified the association between type 2 diabetes and cardiovascular events (p=0.01). The hazard ratio of cardiovascular events associated with the presence of diabetes was higher in women (adjusted hazard ratio, 1.8; 95% CI, 1.2 to 2.7) than in men (adjusted hazard ratio, 1.3; 0.9 to 2). As compared to men without diabetes but with prior cardiovascular disease, risk of cardiovascular events was significantly lower in men with diabetes but without prior cardiovascular disease (adjusted hazard ratio, 0.5; 0.3 to 0.9). In contrast, this risk was equal in women with diabetes but without prior cardiovascular disease and women without diabetes but with prior cardiovascular disease (adjusted hazard ratio, 1.0; 0.6 to 1.7; P for interaction between gender and diabetes=0.05). CONCLUSIONS: Women with diabetes but without prior cardiovascular disease have a risk of cardiovascular events that is similar to that of women without diabetes but with prior cardiovascular disease, whereas in men the presence of prior cardiovascular disease conferred a higher risk. These data emphasise the necessity of aggressive treatment of cardiovascular risk factors in women with type 2 diabetes. PMID- 12909070 TI - From guidelines to clinical practice: the impact of hospital and geographical characteristics on temporal trends in the management of acute coronary syndromes. The Global Registry of Acute Coronary Events (GRACE). AB - AIMS: The extent to which hospital and geographic characteristics influence the time course of uptake of evidence from key clinical trials and practice guidelines is unknown. The gap between evidence and practice is well recognized but the factors influencing this disjunction, and the extent to which such factors are modifiable, remain uncertain. METHODS AND RESULTS: Using chronological data from the GRACE registry (n=12666, July 1999 to December 2001), we test the hypothesis that hospital and geographic characteristics influence the time course of uptake of evidence-based guideline recommendations for acute coronary syndromes (ACS) with and without ST elevation. Certain therapies were widely adopted in both ST-segment elevation myocardial infarction (STEMI) and non ST-segment elevation myocardial infarction (NSTEMI) patients (aspirin >94% of all patients; beta-blockers 85-95%) and changed only modestly over time. Significant increases in the use of low-molecular-weight heparins and glycoprotein IIb/IIIa inhibitors occurred in STEMI and NSTEMI patients in advance of published practice guidelines (September/November 2000) with marked geographical differences. The highest use of LMWH was in Europe in NSTEMI (86.8%) and the lowest in the USA (24.0%). Contrasting geographical variations were seen in the use of percutaneous coronary intervention (PCI) in NSTEMI: 39.5% USA, 34.6% Europe, 33.5% Argentina/Brazil, 25.0% Australia/New Zealand/Canada (July-December 2001). Theuse of PCI was more than five times greater in hospitals with an on-site catheterization laboratory compared to centres without these facilities, and geographic differences remained after correction for available facilities. CONCLUSIONS: Hospital and geographical factors appear to have a marked influence on the uptake of evidence-based therapies in ACS management. The presentation and publication of major international guidelines was not associated with a measurable change in the temporal pattern of practice. In contrast, antithrombotic and interventional therapies changed markedly over time and were profoundly influenced by hospital and geographic characteristics. PMID- 12909072 TI - Impact of rate versus rhythm control on quality of life in patients with persistent atrial fibrillation. Results from a prospective randomized study. AB - AIMS: Despite the high prevalence of atrial fibrillation (AF), there are only limited data on quality of life (QoL) stemming from prospective trials comparing rate versus rhythm control. This prospective study evaluated QoL in patients with symptomatic persistent AF randomized to therapy aiming at rate versus rhythm control. METHODS AND RESULTS: Patients with symptomatic persistent AF (7 to 360 days duration) were prospectively randomized to ventricular rate control (n=125) or to cardioversion and maintenance of sinus rhythm (n=127). QoL was assessed by the Medical Outcomes Study Short Form health survey (SF-36) at baseline and during 1 year of follow-up. Changes in QoL were compared on an intent-to-treat basis, and subsequently between patients in sinus rhythm versus AF. At baseline, all SF-36 scales were reduced compared to healthy controls. At 1 year, six of eight items had improved significantly in patients assigned to rate control, and five of eight items on rhythm control (p=ns). The physical component summary showed a comparable increase with both treatment strategies (rate control: p=0.004; rhythm control: p<0.001) whereas no significant changes were found for the mental component summary. At 1 year, 55% of patients reported a positive health transition with no inter-group differences. There were no significant differences in QoL in patients in sinus rhythm or AF at the end of the observation period. CONCLUSION: In patients with symptomatic persistent AF, the two treatment strategies of rate versus rhythm control are associated with similar improvements in QoL. PMID- 12909071 TI - VErapamil plus antiarrhythmic drugs reduce atrial fibrillation recurrences after an electrical cardioversion (VEPARAF Study). AB - AIMS: To evaluate the impact, on atrial fibrillation (AF) recurrences, of verapamil addition to a class IC or III antiarrhythmic drug in patients, with persistent AF, who underwent an electrical cardioversion (EC). METHODS AND RESULTS: Three hundred sixty-three patients were randomized to receive four different pre-treatment protocols: oral amiodarone (group A), oral flecainide (group F), oral amiodarone plus oral verapamil (group A+V), oral flecainide plus oral verapamil (group F+V). Patients who showed an AF recurrence within 3 months were assigned to the alternative group and underwent a second EC after 48h. During 3 months of follow-up, 89 patients (27.5%) had an AF recurrence. By univariate analysis, verapamil reduced AF recurrences if added to amiodarone or flecainide (from 35% to 20%, P=0.004). Applying Cox proportional hazards regression model, only the younger age, the shorter duration of AF, and the use of verapamil were predictor of maintenance of sinus rhythm after cardioversion. In patients with primary AF recurrence, verapamil addition to group A and F patients, significantly decreased secondary AF recurrence rate as compared to group A+V and F+V patients who stopped the verapamil therapy (68% vs 88%, P=0.03). CONCLUSIONS: The addition of verapamil to class IC or III antiarrhythmic drug significantly reduced the AF recurrences, that were more frequent in older patients and in patients with longer lasting AF; moreover, verapamil was effective in reducing the secondary AF recurrences, too. PMID- 12909073 TI - Regression of left ventricular hypertrophy during 10 years after valve replacement for aortic stenosis is related to the preoperative risk profile. AB - BACKGROUND: Previous studies have suggested that regression of hypertrophy may be the underlying determinant of longevity and left ventricular function after valve replacement (AVR) for aortic stenosis (AS). The potential for hypertrophy regression could therefore be related to the preoperative risk profile. METHODS: Ninety-one consecutive patients with AS had a "project" Doppler-echo and radionuclide ventriculography in addition to the standard investigation programme prior to AVR with a disc valve (19-29mm, n=82), a caged ball valve (26-29mm, n=8), or a stented porcine valve (26mm, n=1); 49 (group A) were selected for a serial follow-up study while 42 served as controls (group B). Forty-two group A patients took part in a 1.5-year examination while 47 (26 group A, 21 group B) patients were studied at 10 years. RESULTS: Groups A and B were comparable as regards all pre- and intra-operative data including left ventricular mass index (LVMi). A previously developed preoperative prognostic index (PI) separated the patients into groups with low (n=23), intermediary (n=19) and high risk (n=49) with 10-year survivals of 87%, 58% and 43% (P<0.01). LVMi dropped from 202+/ 58g/m(2)preoperatively to 152+/-45g/m(2)(P<0.0001) at 1.5 years, and 139+/ 40g/m(2)(P<0.0001) at 10 years (three and six patients, respectively, with paravalvular leak or mitral regurgitation excluded). PI correlated with preoperative (r=0.51, P<0.001), 1.5-year (r=0.46, P<0.01), and 10-year LVMi (r=0.41, P<0.01). Also preoperative left ventricular ejection fraction correlated with the three LVMi measurements. Patients with systemic hypertension had higher LVMi at 1.5 years (193+/-42, n=6 vs 144+/-42, n=33, P<0.05) and 10 years (175+/ 39, n=12 vs 124+/-31g/m(2), n=29, P<0.001). Patients with low, intermediary or high PI, excluding those with hypertension, had 1.5-year LVMi of 110+/-35 (n=8), 134+/-43 (n=9) and 164+/-33g/m(2)(n=16; P<0.01), respectively, and 10-year LVMi of 116+/-25 (n=17), 126+/-27 (n=6), and 146+/-41g/m(2)(n=6; P<0.05), respectively. There was no relation between LVMi at 1.5 or 10 years and peak or mean Doppler gradient, prosthetic valve size, or valve size index. CONCLUSIONS: Left ventricular hypertrophy regression for patients who survived up to 10 years after AVR for AS is dependent on the preoperative risk profile indicating that irreversible myocardial disease is the underlying factor. Systemic hypertension is an important factor in its own right. PMID- 12909074 TI - In-hospital and long-term outcome after sub-massive and massive pulmonary embolism submitted to thrombolytic therapy. AB - BACKGROUND: From a registry of 249 confirmed pulmonary embolism (PE) patients submitted to thrombolytic therapy (TT), we analysed predictors of in-hospital course and long-term mortality. METHODS AND RESULTS: The combined clinical end point of in-hospital course associated death, recurrent PE, repeat thrombolysis, surgical embolectomy or bleeding complications. The long-term follow-up included analysis of survival, and occurrence of PE-related events, defined as recurrent deep vein thrombosis, recurrent PE, occurrence of congestive heart failure or change of New York Heart Association functional class to class III or IV in patients who survived the acute phase.In-hospital clinical course was uneventful in 165 (66.3%) patients. Initial right ventricular (RV) dysfunction was reversible in 80% within 48 h following TT. Initial pulmonary vascular obstruction >70% (RR=5.3 [2.1; 13.6]); haemodynamic instability at presentation (RR=2.6 [1.1; 6]); persistence of septal paradoxical motion after TT (RR=5.9 [1.4; 25.9]); and insertion of intracaval filter (RR=3.7 [1.4; 9.4]) were independent predictors of poor in-hospital course. Mean follow-up was 5.3+/-2.6 years. Of the 227 patients alive after the hospital stay, the probability of survival was 92% at 1 year, 79% at 3 years and 56% at 10 years. Multivariate predictors of long-term mortality were age >75 years (RR=2.73 [2.18; 3.21]; P=0.0002), persistence of vascular pulmonary obstruction >30% after thrombolytic treatment (RR=2.22 [1.69; 2.74]; P=0.003), and cancer (RR=2.03 [1.40; 2.65]; P=0.04). CONCLUSION: The recovery of RV function should be considered as a marker of thrombolysis efficacy, while residual pulmonary vascular obstruction and cancer are independent predictors of long-term mortality. These results advocate the identification of high-risk patients by means of systematic lung-scan and echocardiography pre- and post-thrombolysis, and raise the question of the need for thromboendarterectomy in patients with residual pulmonary vascular obstruction. PMID- 12909075 TI - Celiac disease in patients with sporadic and inherited cardiomyopathies and in their relatives. AB - AIMS: To investigate celiac disease (CD) and related co-morbidity in patients with familial and sporadic cardiomyopathy and in their relatives. METHODS AND RESULTS: We screened anti-human-tissue-transglutaminase (IgA and IgG anti-h-tTG) and anti-endomysial antibodies (AEAs) in 238 consecutive adult patients with inherited or sporadic dilated cardiomyopathy (DCM), 418 relatives, and 2000 healthy blood donors. HLADQ2-DQ8 was tested in tTG-positive subjects. The IgA-tTG positive patients with cardiomyopathy underwent duodenal biopsy. Twenty-six subjects were tTG-positive: five DCM patients (2.1%), two of 28 (7.1%) and three of 390 (0.7%) relatives with and without echocardiographic abnormalities respectively, and 16 controls (0.8%). Twenty-two of 26 subjects were AEA positive, and 25 HLA-positive. Of the five patients with cardiomyopathy and biopsy-proven CD, four suffered iron-deficiency anaemia. Two CD-positive DCM patients and two tTG-positive relatives were from families with inherited disease in which CD did not co-segregate with DCM. CONCLUSIONS; The higher prevalence of CD in patients with sporadic or inherited DCM, and of tTG-positive serology in relatives with echocardiographic abnormalities, suggests that immune-mediated mechanisms are active in subsets of patients/families. However, gluten intolerance cannot be considered causative since CD seems to be associated but not co-segregated with DCM in familial cases. PMID- 12909076 TI - Contrast media as carriers for local drug delivery. Successful inhibition of neointimal proliferation in the porcine coronary stent model. AB - BACKGROUND: Lipophilic taxanes can be dissolved in contrast media at significantly higher concentration than in saline. As contrast media have occasionally been observed to delineate the contour of coronary arteries for some seconds they may serve as a matrix for an antiproliferative drug aimed at preventing restenosis. The aim of this study was to test a novel taxane-contrast agent formulation for this new approach in the setting of coronary stenting. METHODS AND RESULTS: In cell culture experiments (bovine vascular smooth muscle cells), 60-min incubation with contrast agent-taxane formulations (iopromide paclitaxel, iopromide-protaxel) induced a significant, concentration-dependent inhibition of vascular smooth muscle cell (VSMC) proliferation over 12 days. Shorter incubation times of 10 and 3 min showed the same efficacy. For in vivo investigation, 16 stents were implanted into the coronary arteries of eight pigs using a 1.3 to 1 overstretch ratio. A control group received iopromide 370 alone while the treatment group was injected with a iopromide-protaxel formulation at a dose of 74 micromol/l, which is far below protaxel levels inducing systemic toxicity. Quantitative angiography and histomorphometry of the stented arteries asserted statistic equality of the baseline parameters between the control and treatment groups. After 28 days, the treatment group showed a marked reduction of the parameters characterizing in-stent restenosis, especially a 34% reduction of the neointimal area. CONCLUSIONS: First evidence is provided that using a contrast agent as solvent for a taxane constitutes a new drug delivery mechanism able to inhibit in-stent restenosis in the porcine restenosis model. PMID- 12909077 TI - Mechanisms of neuronal cell death: diverse roles of calcium in the various subcellular compartments. PMID- 12909078 TI - Neuronal ageing from an intraneuronal perspective: roles of endoplasmic reticulum and mitochondria. AB - The nature of brain ageing and the age-dependent decline in cognitive functions remains poorly understood. Physiological brain ageing is characterised by mild mental dysfunctions, whereas age-dependent neurodegeneration, as illustrated by Alzheimer disease (AD), results rapidly in severe dementia. These two states of the aged brain, the physiological and the pathological, are fundamentally different as the latter stems from significant neuronal loss, whereas the former develops without significant neuronal demise. In this paper, we review the changes in neuronal Ca(2+) homeostasis that occur during brain ageing, and conclude that normal, physiological ageing is characterised mainly by a decrease of neuronal homeostatic reserve, defined as the capacity to respond effectively to functional and metabolic stressors, but does not reach the trigger required to induce neuronal death. In contrast, during neurodegenerative states, Ca(2+) homeostasis is affected early during the pathological process and result in significant neuronal demise. We also review recent evidence suggesting that the endoplasmic reticulum (ER) might play an important role in controlling the balance between healthy and pathological neuronal ageing. PMID- 12909079 TI - Molecular mechanisms of calcium-dependent neurodegeneration in excitotoxicity. AB - Excitotoxicity contributes to neuronal degeneration in many acute CNS diseases, including ischemia, trauma, and epilepsy, and may also play a role in chronic diseases, such as amyotrophic lateral sclerosis (ALS). Key mediators of excitotoxic damage are Ca ions (Ca(2+)), which under physiological conditions govern a multitude of cellular processes, including cell growth, differentiation, and synaptic activity. Consequently, homeostatic mechanisms exist to maintain a low intracellular Ca(2+) ion concentration so that Ca(2+) signals remain spatially and temporally localized. This permits multiple independent Ca-mediated signaling pathways to occur in the same cell. In excitotoxicity, excessive synaptic release of glutamate can lead to the disregulation of Ca(2+) homeostasis. Glutamate activates postsynaptic receptors, including the ionotropic N-methyl-D-aspartate (NMDA), 2-amino-3-(3-hydroxy-5-methylisoxazol-4-yl) proprionate (AMPA), and kainate receptors. Upon their activation, these open their associated ion channel to allow the influx of Ca(2+) and Na(+) ions. Although physiological elevations in intracellular Ca(2+) are salient to normal cell functioning, the excessive influx of Ca(2+) together with any Ca(2+) release from intracellular compartments can overwhelm Ca(2+)-regulatory mechanisms and lead to cell death. Although Ca(2+) disregulation is paramount to neurodegeneration, the exact mechanism by which Ca(2+) ions actually mediate excitotoxicity is less clear. One hypothesis outlined in this review suggests that Ca(2+)-dependent neurotoxicity occurs following the activation of distinct signaling cascades downstream from key points of Ca(2+) entry at synapses, and that triggers of these cascades are physically co-localized with specific glutamate receptors. Thus, we summarize the importance of Ca(2+) regulation in mammalian neurons and the excitotoxicity hypothesis, and focus on the molecular determinants of glutamate receptor-mediated excitotoxic mechanisms. PMID- 12909080 TI - Capacitative calcium entry in the nervous system. AB - Capacitative calcium entry is a process whereby the depletion of Ca(2+) from intracellular stores (likely endoplasmic or sarcoplasmic reticulum) activates plasma membrane Ca(2+) channels. Current research has focused on identification of capacitative calcium entry channels and the mechanism by which Ca(2+) store depletion activates the channels. Leading candidates for the channels are members of the transient receptor potential (TRP) superfamily, although no single gene or gene product has been definitively proven to mediate capacitative calcium entry. The mechanism for activation of the channels is not known; proposals fall into two general categories, either a diffusible signal released from the Ca(2+) stores when their Ca(2+) levels become depleted, or a more direct protein-protein interaction between constituents of the endoplasmic reticulum and the plasma membrane channels. Capacitative calcium entry is a major mechanism for regulated Ca(2+) influx in non-excitable cells, but recent research has indicated that this pathway plays an important role in the function of neuronal cells, and may be important in a number of neuropathological conditions. This review will summarize some of these more recent findings regarding the role of capacitative calcium entry in normal and pathological processes in the nervous system. PMID- 12909081 TI - Calcium dynamics and endoplasmic reticular function in the regulation of protein synthesis: implications for cell growth and adaptability. AB - The endoplasmic reticulum (ER) possesses the structural and functional features expected of an organelle that supports the integration and coordination of major cellular processes. Ca(2+) sequestered within the ER sustains lumenal protein processing while providing a reservoir of the cation to support stimulus-response coupling in the cytosol. Release of ER Ca(2+) sufficient to impair protein processing promotes ER stress and signals the "unfolded protein response" (UPR). The association of the UPR with an acute suppression of mRNA translational initiation and a longer term up-regulation of ER chaperones and partial translational recovery is discussed. Regulatory sites in mRNA translation and the mechanisms responsible for the early and later phases of the UPR are reviewed. The regulatory significance of GRP78/BiP, a multifunctional, broad-specificity ER chaperone, in the coordination of ER protein processing with mRNA translation during acute and chronic ER stress is addressed. The relationship of ER stress to protein misfolding in the cytoplasm is examined. Translational alterations in embryonic cardiomyocytes during treatments with various Ca(2+)-mobilizing, growth promoting stimuli are described. The importance of ER Ca(2+) stores, ER chaperones, and cytosolic-free Ca(2+) in translational control and growth promotion by these stimuli is assessed. Some perspectives are provided regarding Ca(2+) as an integrating factor in the generation or diversion of metabolic energy. Circumstances impacting upon cellular adaptability during exposure to growth stimuli or during stressful conditions that require rapid adjustments in ATP for continued viability are considered. PMID- 12909082 TI - Endoplasmic reticulum: a primary target in various acute disorders and degenerative diseases of the brain. AB - Changes in neuronal calcium activity in the various subcellular compartments have divergent effects on affected cells. In the cytoplasm and mitochondria, where calcium activity is normally low, a prolonged excessive rise in free calcium levels is believed to be toxic, in the endoplasmic reticulum (ER), in contrast, calcium activity is relatively high and severe stress is caused by a depletion of ER calcium stores. Besides its role in cellular calcium signaling, the ER is the site where membrane and secretory proteins are folded and processed. These calcium-dependent processes are fundamental to normal cell functioning. Under conditions of ER dysfunction unfolded proteins accumulate in the ER lumen, a signal responsible for activation of the unfolded protein response (UPR) and the ER-associated degradation (ERAD). UPR is characterized by activation of two ER resident kinases, PKR-like ER kinase (PERK) and IRE1. PERK induces phosphorylation of the eukaryotic initiation factor (eIF2alpha), resulting in a shut-down of translation at the initiation step. This stress response is needed to block new synthesis of proteins that cannot be correctly folded, and thus to protect cells from the effect of unfolded proteins which tend to form toxic aggregates. IRE1, on the other hand, is turned after activation into an endonuclease that cuts out a sequence of 26 bases from the coding region of xbp1 mRNA. Processed xbp1 mRNA is translated into the respective protein, an active transcription factor specific for ER stress genes such as grp78. In acute disorders and degenerative diseases, the ER calcium pool is a primary target of toxic metabolites or intermediates, such as oxygen free radicals, produced during the pathological process. Affected neurons need to activate the entire UPR to cope with the severe form of stress induced by ER dysfunction. This stress response is however hindered under conditions where protein synthesis is suppressed to such an extent that processed xbp1 mRNA is not translated into the processed XBP1 protein (XBP1(proc)). Furthermore, activation of ERAD is important for the degradation of unfolded proteins through the ubiquitin/proteasomal pathway, which is impaired in acute disorders and degenerative diseases, resulting in further ER stress. ER functioning is thus impaired in two different ways: first by the direct action of toxic intermediates, produced in the course of the pathological process, hindering vital ER reactions, and second by the inability of cells to fully activate UPR and ERAD, leaving them unable to withstand the severe form of stress induced by ER dysfunction. PMID- 12909083 TI - Neuronal and glial calcium signaling in Alzheimer's disease. AB - Cognitive impairment and emotional disturbances in Alzheimer's disease (AD) result from the degeneration of synapses and death of neurons in the limbic system and associated regions of the cerebral cortex. An alteration in the proteolytic processing of the amyloid precursor protein (APP) results in increased production and accumulation of amyloid beta-peptide (Abeta) in the brain. Abeta has been shown to cause synaptic dysfunction and can render neurons vulnerable to excitotoxicity and apoptosis by a mechanism involving disruption of cellular calcium homeostasis. By inducing membrane lipid peroxidation and generation of the aldehyde 4-hydroxynonenal, Abeta impairs the function of membrane ion-motive ATPases and glucose and glutamate transporters, and can enhance calcium influx through voltage-dependent and ligand-gated calcium channels. Reduced levels of a secreted form of APP which normally regulates synaptic plasticity and cell survival may also promote disruption of synaptic calcium homeostasis in AD. Some cases of inherited AD are caused by mutations in presenilins 1 and 2 which perturb endoplasmic reticulum (ER) calcium homeostasis such that greater amounts of calcium are released upon stimulation, possibly as the result of alterations in IP(3) and ryanodine receptor channels, Ca(2+) ATPases and the ER stress protein Herp. Abnormalities in calcium regulation in astrocytes, oligodendrocytes, and microglia have also been documented in studies of experimental models of AD, suggesting contributions of these alterations to neuronal dysfunction and cell death in AD. Collectively, the available data show that perturbed cellular calcium homeostasis plays a prominent role in the pathogenesis of AD, suggesting potential benefits of preventative and therapeutic strategies that stabilize cellular calcium homeostasis. PMID- 12909084 TI - Ca(2+) signalling in mitochondria: mechanism and role in physiology and pathology. AB - Over recent years, a renewed interest in mitochondria in the field of Ca(2+) signalling has highlighted their central role in regulating important physiological and pathological events in animal cells. Mitochondria take up calcium through an uptake pathway that, due to its low-Ca(2+) affinity, demands high local calcium concentrations to work. In different cell systems high-Ca(2+) concentration microdomains are generated, upon cell stimulation, in proximity of either plasma membrane or sarco/endoplasmic reticulum Ca(2+) channels. Mitochondrial Ca(2+) accumulation has a dual role, an universal one, which consists in satisfying energy demands by increasing the ATP production through the activation of mitochondrial enzymes, and a cell type specific one, which, through the modulation of the spatio-temporal dynamics of calcium signals, contributes to modulate specific cell functions. Recent work has revealed the central role of mitochondria dysfunction in determining both necrotic and apoptotic cell death. Evidence is also accumulating that suggests that alterations in mitochondrial function may act as predisposing factors in the pathogenesis of a number of neurodegenerative disorders. These include inherited disorders of the mitochondrial genome in which a defect in mitochondrial calcium accumulation has been shown to correlate with a defect in ATP production, thus suggesting a possible involvement of mitochondrial Ca(2+) dysfunction also for this group of diseases. This review analyses recent developments in the area of mitochondrial Ca(2+) signalling and attempts to summarise cell physiology and cell pathology aspects of the mitochondrial Ca(2+) transport machinery. PMID- 12909085 TI - Interactions between mitochondrial bioenergetics and cytoplasmic calcium in cultured cerebellar granule cells. AB - The mitochondrion has moved to the center stage in the drama of the life and death of the neuron. The mitochondrial membrane potential controls the ability of the organelle to generate ATP, generate reactive oxygen species and sequester Ca(2+) entering the cell. Each of these processes interact, and their deconvolution is far from trivial. The cultured cerebellar granule cell provides a model in which knowledge gained from studies on isolated mitochondria can be applied to study the role played by the organelles in the maintenance of Ca(2+) homeostasis in the cell under resting, stimulated and pathophysiological conditions. In particular, mitochondria play a complex role in the response of the neuron to excitotoxic stimulation of NMDA and AMPA-kainate selective glutamate receptors. One goal of research in this area is to provide clues as to possible ways in which modulators of mitochondrial function may be used as neuroprotective agents, since mitochondrial Ca(2+) accumulation seems to play a key role in glutamate excitotoxicity. PMID- 12909086 TI - Ca(2+)/CREB/CBP-dependent gene regulation: a shared mechanism critical in long term synaptic plasticity and neuronal survival. AB - CREB is a transcription factor critical for long-term synaptic plasticity. Intriguingly, recent work has elucidated a role for CREB, as well as upstream CREB kinases, in the control of activity-dependent neuronal survival. Additionally, analysis of the molecular pathology of polyglutamine-repeat diseases suggest that alteration of pCREB-CBP function may underlie, at least in part, the neurodegenerative process. Taken together, these new findings support the idea that Ca(2+)/CREB/CBP-dependent gene regulation might be a shared mechanism critical in both long-term synaptic plasticity and neuronal survival. PMID- 12909087 TI - Studies of the thermal degradation of waste rubber. AB - One kind of Chinese waste tire's sample was pyrolyzed under an inert atmosphere by using thermo-gravimetric apparatus (TGA) and differential thermal analysis (DTA). Different pyrolysis temperature ranges were determined according to the reaction transition temperature obtained by TGA and DTA. Then, at each temperature range, the pyrolysis gaseous products were analyzed by gas chromatography (GC). The influence of the temperature range on the relative yields of the major decomposition products is studied, and a mechanism for the formation of the main components was also investigated. The results indicate that pyrolysis of waste rubber follows the radical mechanism, and the major products are not seriously affected by increasing the temperature from room temperature to 420 degrees C and from 421 to 600 degrees C, but the degradation of blend rubber is different for each of the compositional elastomer. PMID- 12909088 TI - Hydrological performance of MSW incineration residues and MSW co-disposed with sludge in full-scale cells. AB - Water flows were analysed for the filling phase and the first 4 years after closure of two types of full-scale landfill cells: 'special cells' containing mostly fly ash from municipal solid waste (MSW) incineration disposed with other special/hazardous waste, and 'biocells' (biological cells) containing co-disposed MSW and food industry sludge. The landfill cells were constructed about -1.5 m above sea level (masl) at Lomma Bay, southern Sweden. The hydrological effects of water intrusion into the special cells from surroundings and sludge moisture within the biocells were studied. HELP modelling of hydrological processes predicted delay in peaks of leachate generation from uncovered special cells following rain, which was not confirmed. Faster leachate production as a response to rainfall from special cells than from biocells was observed. It was inferred that special waste has more intensive channelling, lower water absorption and higher hydraulic conductivity than mixtures of sludge/MSW. To avoid convergence problems in modelling uncovered special cells, the use of a 5 cm deep top layer with saturated hydraulic conductivity 1.7 x 10(-3) cm s(-1), porosity 0.437, and field capacity 0.105, is suggested. PMID- 12909089 TI - Study of the presence of fluorine in the recycled fractions during carbothermal treatment of EAF dust. AB - Carbothermal treatment tests of electric arc furnace dusts (EAFD) using the Waelz kiln process were carried out in pilot-scale for the production of zinc oxide. The association of halides in the EAFD, and the recycled products, such as zinc oxide fumes and high-grade iron contents fractions were examined by X-ray diffraction (XRD), scanning electron microscope (SEM) and transmission electron microscope (TEM) analysis. XRD reveals the presence of chlorine and fluorine in the dusts in the form of KCl, NaCl and CaF2. An ultra-pure fraction of zinc was obtained after the Double Leaching Waelz Oxide (DLWO) process was performed on the zinc oxide fumes. The halide contents were reduced to approximately 100 ppm Cl and 700 ppm F. The rest of these elements are in the form of CaF2. About 65% F is volatilised as lead and zinc fluorides, 15% is expected in the magnetic fractions and 20% in non-magnetic fractions as CaF2 and MnF2, respectively. PMID- 12909090 TI - The simulation of condensation removal of a heavy metal from exhaust gases onto sorbent particles. AB - A numerical model BAEROSOL for solving the general dynamic equation (GDE) of aerosols is presented. The goal was to model the capture of volatilized metals by sorbents under incinerator-like conditions. The model is based on algorithms presented by Jacobson and Turco [Aerosol Science and Technology 22 (1995) 73]. A hybrid size bin was used to model growth and formation of particles from the continuum phase and the coagulation of existing particles. Condensation and evaporation growth were calculated in a moving size bin approach, where coagulation and nucleation was modeled in the fixed size bin model of the hybrid grid. To account for the thermodynamic equilibrium in the gas phase, a thermodynamic equilibrium code CET89 was implemented. The particle size distribution (PSD) calculated with the model was then compared to analytical solutions provided for growth, coagulation and both combined. Finally, experimental findings by Rodriguez and Hall [Waste Management 21 (2001) 589-607] were compared to the PSD predicted by the developed model and the applicability of the model under incineration conditions is discussed. PMID- 12909091 TI - Advanced ash management technologies for CFBC ash. AB - The combustion of high-sulphur coal demands the reduction of sulphur emissions. The sorbent most often used in sulphur capture technology is calcium-based. Ashes from technologies such as circulating fluidized bed combustion (CFBC), therefore, contain high calcium levels. The use and disposal of these ashes poses challenges, because of highly exothermic reactions with water, high-pH leachates, and excessive expansion of solidified materials. This paper looks at the potential of two post-combustion ash treatment processes, CERCHAR hydration and AWDS disposal, in solving these challenges. A high-sulphur coal-derived CFBC ash is examined, after CERCHAR hydration treatment, in conjunction with a conventionally hydrated ash, in a range of chemical, geotechnical and utilization scenarios. The ashes are used to make no-cement and roller-compacted concrete as well as Ash Water Dense Suspensions (AWDS). The solidified mortar paste from no cement concrete is subjected to an extensive geochemical examination to determine how solidification progresses and strength develops, from a chemical point of view. PMID- 12909092 TI - Characterisation of Malian and Belgian solid waste composts with respect to fertility and suitability for land application. AB - Two composts, a Malian (C1) and a Belgian (C2), and a peat substrate (C3) were analyzed for their suitability for land application. The results revealed that the materials can be used in agriculture but only the application of the peat substrate may create N immobilization. Composts had higher P, Ca, and Mg contents and lower C and K contents and C/N ratio than the peat substrate. The available P extracted from the three materials ranged from 15% (for C2) to 48% (for C3) and available K from 36% (in the composts) to 48% (for C3) of the total elements respectively. The fractionation of Mn, Fe, Zn and Cu in operationally defined fractions (water soluble, exchangeable, complexed, organically bound and residual) allows an estimation of the availability of metals for uptake. For example, 16% of the total Mn in the composts (C1 and C2) and 22% in the peat substrate would be plant available. Available Fe in the three materials was less than 2%. Available Zn varied between 10 and 25%. The fractionation allowed an estimation of the potential for contamination of groundwater following the applications of composts to agricultural lands. PMID- 12909093 TI - Polyalkylphenyl-sulphonic acid with acid groups of variable strength from compost. AB - The insoluble organic fraction (humin-like material, HLM) from rice hull-dairy cattle compost is well converted into water soluble HLM-sulphonate by reaction in liquid SO3. Microanalytical, potentiometric, molecular weight, and NMR data are consistent with a highly homogeneous polymeric arylsulphonate having 4000 Da MW, 1.3 sulphonic groups per aromatic ring and significant content of carboxylic and phenolic groups. By comparison with structure-property relationships for commercial lignosulphonates derived from the pulp and paper industry, the above arylsulphonate is likely to be a candidate at a variety of applications in the chemical industry and in agriculture. Therefore, sulphonation is a means for upgrading composts HLM to the same uses as for commercial lignosulphonates. PMID- 12909094 TI - Removal of copper, chromium, and arsenic from CCA-C treated wood by EDTA extraction. AB - Ethylenediaminetetracetic acid (EDTA) is one of the most common chelators used to bind the metal ions in extremely stable complexes in heavy metal contaminated soils and thus to remediate such substrates. EDTA forms water soluble complexes with many metal ions and it is used to release the various metals. In this study, EDTA extraction of copper, chromium, and arsenic from chromated copper arsenate (CCA-C) treated wood was evaluated using batch leaching experiments. CCA-treated wood samples were extracted with eight different concentrations of EDTA for 4, 8, 18, and 24 h at room temperature. Exposing CCA-treated chips and sawdust to EDTA extraction enhanced removal of CCA components compared with extraction by deionized water. Grinding CCA-treated wood chips into 40-mesh sawdust provided greater access to and removal of CCA components. Extraction with 1% EDTA solution for 24 h removed 60% copper, 13% chromium, and 25% arsenic from treated chips. EDTA extraction of treated sawdust samples resulted in 93% copper, 36% chromium, and 38% arsenic removal. CCA leaching from treated wood blocks was also evaluated according to modified AWPA E11-99 standard test method of determining the leachability of wood preservatives. Leaching of CCA components from treated wood blocks with 1% EDTA solution for 14 days caused more copper leaching compared to leaching with deionized water. Leaching with 1% EDTA for 14 days removed 53% copper from the blocks whereas 14% copper was leached from the blocks with deionized water. The results suggest that EDTA extraction removes significant quantities of copper from CCA-treated wood. Thus, EDTA could be important in the remediation of wood waste treated with the newest formulations of organometalic copper compounds and other water-borne wood preservatives containing copper. PMID- 12909095 TI - Criteria selection for landfills: do we need a limitation on inorganic total content? AB - The chemistry of the landfill environment is complex and dissolution rates may be higher than in natural systems. As will be shown in this paper, there are often great uncertainties in terms of leaching kinetics and long-term emissions of landfills in general. Leaching tests used in the laboratory may reflect a leaching behaviour not consistent with real landfill scenarios. Geochemical calculations for model systems provide an impression of the possible long-term behaviour of buffered, inorganic landfills. For a buffered carbonate-containing system (this system may be a mono-landfill of slag and/or ash after a few decades and/or centuries), maximums for the solubility of the elements Zn, Cd, Pb and Cu can be calculated while taking into account the presence of gypsum, the complexation in water ("speciation") and other factors influencing solubility. We have made these calculations using the recent version of the geochemical code PHREEQC-2. Finally, we conclude that under most circumstances the total elemental content, or the so-called "total availability," is a valuable assessment tool when combined with other criteria, such as leachable amounts and leachate test results. Exceptions may be carbonate-buffered monofill scenarios (e.g. monofill or quasi-monofill of MSWI residues), where setting limits on some elemental contents (Cu, Zn, Fe and Mn) does not seem to be very useful. PMID- 12909097 TI - Clay particle retention in small constructed wetlands. AB - Constructed wetlands (CWs) can be used to mitigate non-point source pollution from arable fields. Previous investigations have shown that the relative soil particle retention in small CWs increases when hydraulic load increases. This paper investigates why this phenomenon occurs, even though common retention models predict the opposite, by studying clay and silt particle retention in two Norwegian CWs. Retention was measured with water flow proportional sampling systems in the inlet and outlet of the wetlands, and the texture of the suspended solids was analyzed. The surface area of the CWs was small compared to the watershed area (approximately 0.07%), giving high average hydraulic loads (1.1 and 2.0 md(-1)). One of the watersheds included only old arable land, whereas the other included areas with disturbed topsoil after artificial land leveling. Clay particle retention was 57% for the CW in the first watershed, and 22% for the CW in the disturbed watershed. The different behavior of the wetlands could be due to differences in aggregate size and stability of the particles entering the wetlands. Results showed that increased hydraulic loads did affect CW retention negatively. However, as runoff increased, soil particles/aggregates with higher sedimentation velocities entered the CWs (e.g., the clay particles behaved as silt particles). Hence, clay particle settling velocity is not constant as assumed in many prediction models. The net result was increased retention. PMID- 12909098 TI - Kinetics of 4-nitrophenol biodegradation in a sequencing batch reactor. AB - In this paper, the biodegradation process of 4-nitrophenol (4NP) in a sequencing batch reactor has been investigated. Kinetic tests have been carried out on biomass grown on mixed substrate (4NP plus biogenic substrate) both in the presence of a biogenic substrate fraction in the feed and with 4NP as the sole carbon source. Removal kinetics for all tests is well described by the typical substrate inhibition pattern as predicted by the Haldane equation. In both sets, estimated kinetic parameters are very similar: no beneficial effect of the biogenic fraction is observed on the 4NP removal while increasing trend of 4NP maximum removal rate with the 4NP/COD(TOT) ratio in the feed has been observed. This finding has been modelled by estimating the fraction of the total biomass involved in 4NP biodegradation as a function of 4NP concentration in the feed. High removal rates, short acclimation times and good settling characteristics of produced sludge (observed during the whole working period) confirm the suitability of periodic systems in enhancing the bacterial potentialities for biodegradation of xenobiotic compounds. PMID- 12909099 TI - Application of nano TiO(2) towards polluted water treatment combined with electro photochemical method. AB - A novel composite reactor was prepared and studied towards the degradation of organic pollutants in this work. In the reactor, a UV lamp was installed to provide energy to excite nano TiO(2), which served as photocatalyst, leading to the production of hole-electron pairs, and a three-electrode electrolysis system was used to accumulate H(2)O(2) which played an important role in the degradation process. The reactor was evaluated by the degradation process of rhodamine 6G (R 6G), and the data obtained in the experiments showed that the combination of the photochemical and electrochemical system raised the degradation rate of R-6G greatly; the working mechanism of the reactor was also discussed in the article. The prepared reactor was also utilized to treat polluted water from dyeing and printing process. After continuous treatment for 0.5h, chemical oxygen demand biochemical oxygen demand, quantity of bacteria and ammonia nitrogen of the polluted water were reduced by 93.9%, 87.6%, 99.9% and 67.5%, respectively, which indicated that the method used here could be used for effective organic dyes degradation. PMID- 12909100 TI - Comparison and evaluation of empirical zone settling velocity parameters based on sludge volume index using a unified settling characteristics database. AB - Over the past few decades classic and contemporary research on the process of secondary clarification in activated sludge plants has illuminated several aspects of the solids-flux theory which actually require a closer examination. Inter alia the most challenging and controversial part in the field has been the development of reliable models for the settling properties of the activated sludge in the settler. Numerous studies have been performed aiming at the evolvement of reliable mathematical formulas that would satisfactorily describe this behaviour, but no universally accepted solution seems to exist to date. That is mostly because different experimental conditions, sludge types and instrumentation have been employed, thus complicating the process of reaching a conclusive result. In order to bring theoretical and practical developments of secondary settling tank design and simulation closer together, a number of related tasks are addressed in this study by the use of an integrated and unified settling characteristics database. Several drawbacks and advantages of the methodologies published hitherto are examined on a universal basis and under the same assumptions in order to reveal artifacts that complicate the procedure of settling velocity estimation. It is suggested that universally accepted solutions may be feasible especially for design purposes. For simulation analysis real-time data of settling velocity should be tested rather than values derived from laboratory experiments which are shown to produce different results depending on the applied approach. In conclusion, an integrated database is proposed as a means for a more robust and universally accepted design procedure. PMID- 12909101 TI - Development of a new method for the determination of nitrosamines by micellar electrokinetic capillary chromatography. AB - A new method was developed for separation and quantification of nitrosamines by micellar electrokinetic capillary chromatography (MEKC). The effects of composition of the buffer, concentration of sodium dodecyl sulfate (SDS), gamma cyclodextrin (CD) and pH on the separation and migration times of the nitrosamines were investigated. The different instrumental variables affecting sensitivity and resolution, such as power supply, injection mode and wavelength for measurement, were carefully optimized. The direct application of this method to aqueous synthetic samples allowed the separation and determination of nitrosamine mixtures at the mg x L(-1) level. The calibration curves revealed a linear range between 0.25 and 10 mg x L(-1), with detection limit (DL) varying from 0.16 to 0.27 mg x L(-1), quantification limit (QL) varying from 0.52 to 0.90 mg x L(-1), and RSD relative standard deviation (RSD) from 4.5% to 8.5%. The proposed method was successfully applied to the determination of dimethylnitrosamine (DMN), diethylnitrosamine (DEN), N-nitrosopyrrolidine (NPYR), N-nitrosomorpholine (NMOR) and N-nitrosopiperidine (NPIP). PMID- 12909102 TI - The mechanism of metals precipitation by biologically generated alkalinity in biofilm reactors. AB - Microbial strains, characterized by increased tolerance and ability to grow in metal bearing wastewaters as well as by effective metal sequestering capability by both active (bioaccumulative) and passive (biosorptive) processes, were tested as inoculum for metal laden wastewater treatment systems. Their capacity to grow in metal bearing wastewater, using an easily available and inexpensive carbon source such as acetate, was studied in batch experiments. Two principal conclusions were drawn: (1). Growth was observed for all the strains examined suggesting that the strains can be acclimated to metals bearing wastewaters. (2). Solution pH increased from neutral to alkaline values during growth (pH(initial)=7, pH(final)=10). The later was observed systematically for all strains. Metal precipitation, due to the metabolically generated alkalinity is expected as a result.Supporting evidence for this hypothesis was provided during the operation of two pilot moving bed sand filters treating two different metal bearing wastewaters. Acetate was used as carbon source to support the growth and maintenance of microbial biomass on the sand grains of the filters. The characteristics of the sludge produced from the operation of the pilot plants were subsequently studied in the laboratory. Both sludges were significantly loaded with large amounts of metals.A mechanism of metal precipitation induced by the metabolically generated alkalinity, when acetate is used as carbon source, could be proposed as the main process responsible for the metals sequestering inside the moving bed sandfilter reactor. PMID- 12909103 TI - Possible cause of excess sludge reduction in an oxic-settling-anaerobic activated sludge process (OSA process). AB - Modification of a conventional activated sludge process by inserting a sludge holding tank in a sludge return line forms an oxic-settling-anaerobic (OSA) process that may provide a cost-effective way to reduce excess sludge production in activated sludge processes. In this paper we systematically evaluate the following possible scenarios that may explain the reduction of excess sludge in the OSA process: (i). energy uncoupling, (ii). domination of slow growers, (iii). soluble microbial products (SMPs) effect and (iv). sludge decay in the sludge holding tank under a low oxidation-reduction potential (ORP) condition. Results show that only the final scenario may reasonably explain this reduction. It has also been found that the sludge decay process in the sludge holding tank may involve the reduction of the cell mass. PMID- 12909104 TI - Electrodialysis and nanofiltration of surface water for subsequent use as infiltration water. AB - In order to achieve stable groundwater levels, an equilibrium between the use of groundwater for drinking water production and natural or artificial groundwater recharge by infiltration is needed. Local governments usually require that the composition of the water used for artificial recharge is similar to the surface water that is naturally present in the specific recharge area. In this paper, electrodialysis (ED) and nanofiltration were evaluated as possible treatment technologies for surface water from a canal in Flanders, the North of Belgium, in view of infiltration at critical places on heathlands. Both methods were evaluated on the basis of a comparison between the water composition after treatment and the composition of local surface waters. The treatment generally consists of a tuning of pH and the removal of contaminants originating from industrial and agricultural activity, e.g., nitrates and pesticides. Further evaluation of the influence of the composition of the water on the characteristics of the artificial recharge, however, was not envisaged. In a case study of water from the canal Schoten-Dessel, satisfactory concentration reductions of Cl(-), SO(4)(2-), NO(3)(-), HCO(3)(-), Na(+), Mg(2+), K(+) and Ca(2+) were obtained by ultrafiltration pretreatment followed by ED. Nanofiltration with UTC-20, N30F, Desal 51 HL, UTC-60 and Desal 5 DL membranes resulted in an insufficient removal level, especially for the monovalent ions. PMID- 12909105 TI - Comparison of effectiveness of different ashing auxiliaries for determination of phosphorus in natural waters, aquatic organisms and sediments by ignition method. AB - An improved method for the determination of phosphorus in natural waters, aquatic organisms and sediments by ignition method is proposed. The recoveries of phosphorus (P) from selected inorganic and organic P-containing compound standards after ignition with different auxiliaries, such as MgSO(4), Mg(NO(3))(2), MgCl(2), Mg(Ac)(2) and CaCl(2), were compared. We found that the phosphorus from most compound standards could not be completely recovered when these compounds were ignited (450-500 degrees C) with the MgSO(4) as auxiliary and the baked residue was extracted with 0.2 mol x l(-1) HCl for 30 min at 80 degrees C or at room temperature. P recoveries, for example, were poor, less than 85%, if pyrophosphate and metaphosphate were ignited with the addition of MgSO(4) prior to the extraction of the baked residue with 0.2 mol x l(-1) HCl at 80 degrees C for 30 min. In contrast, MgCl(2), Mg(Ac)(2) and CaCl(2), as well as Mg(NO(3))(2), could all yield complete P recoveries at routine ashing temperatures (450-500 degrees C). The results demonstrate that MgCl(2) is a more effective auxiliary agent for the determination of phosphorus in natural waters, aquatic organisms and sediments by ignition method than MgSO(4) which is commonly used. PMID- 12909106 TI - Removal of copper ions from aqueous solutions by a steel-making by-product. AB - A study is made of the use of a steel-making by-product (rolling mill scale) as a material for removing Cu(2+) ions from aqueous solutions. The influence of contact time, initial copper ion concentration and temperature on removal capability is considered. The removal of Cu(2+) ions from an aqueous solution involves two processes: on the one hand, the adsorption of Cu(2+) ions on the surface of mill scale due to the iron oxides present in the latter; and on the other hand, the cementation of Cu(2+) onto metallic iron contained in the mill scale. Rolling mill scale is seen to be an effective material for the removal of copper ions from aqueous solutions. PMID- 12909107 TI - Triclosan in a sewage treatment process--balances and monitoring data. AB - In a German sewage treatment plant that processes 200000 m(3) wastewater per day, the concentrations of 2,4,4'-trichloro, 2'-hydroxy-phenylether (triclosan) in the in-flowing ( approximately 1000 ng x l(-1)) as well as in the out-flowing water ( approximately 50 ng x l(-1)) are compared to the concentrations measured in sludge (1200 ng x g(-1)). Considering the mass flow of water and sludge in the respective plant, balances including water and sludge are calculated. Thirty percent of the triclosan is sorbed with weak bonds to the sludge, while some amounts are sorbed as bound residues in the sludge. About 5% is dissolved in the out-flowing water. Thus most of the in-flowing material is not recovered as the parent compound but it is likely that it is transformed to other metabolites or unrecovered bound residues. These data are compared to the monitoring of sewage sludge of 20 different plants in this region, most of which are smaller, though. The concentrations found in these sludges vary from 1000-8000 ng x g(-1). PMID- 12909108 TI - Phenolic removal in a model olive oil mill wastewater using Pleurotus ostreatus in bioreactor cultures and biological evaluation of the process. AB - Pleurotus ostreatus grown in bioreactor batch cultures in a model phenolic wastewater (diluted and sterilized olive oil mill wastewater-OMW), caused significant phenolic removal. Laccase, the sole ligninolytic enzyme detected in the growth environment, was produced during primary metabolic growth. The bioprocess was simulated with the aid of a mathematical model and the parameters of growth were determined. When the fungal biomass was increased in the reactor (during repeated batch experiments) the rate of reducing sugars consumption progressively increased, but a phenolic fraction seemed of being strongly resistant to oxidation. The toxicity of OMW against the seeds of Lepidium sativum and the marine Branchiopoda Artemia sp. was significantly decreased after biotreatment. On the contrary, the toxicity against the freshwater Branchiopoda Daphnia magna was not affected by the treatment, whereas on the soil and freshwater sediments Ostracoda Heterocypris incongruens was slightly decreased. Both treated and untreated OMWs, used as water for irrigation of lettuce and tomato plants, did not significantly affect the uptake of several nutrients by the cultivated plants, but resulted in a decrease in the plant yields, which was minimized when high OMW dilutions were used. As a conclusion, P. ostreatus is able to reduce phenolic content and toxicity of sterilized OMW, in bioreactor cultures. However, high OMW dilutions should be used, and/or additional treatment should be applied before use of the OMW in the environment, e.g. as water for irrigation. Further research should be done in order to transfer this technology under industrial conditions (e.g. by using unsterilized OMW). PMID- 12909109 TI - Mechanism of lead adsorption from aqueous solutions using an adsorbent synthesized from natural condensed tannin. AB - Adsorption is a method for removing lead from wastewater. The adsorption of lead on a new adsorbent synthesized from natural condensed tannin has been investigated using a series of batch adsorption experiments. The study on the adsorption mechanism indicates that the adsorbent performed in aqueous solutions as an ionic exchanger whose end group was sodium ion (Na(+)). One lead (II) ion (Pb(2+)) was adsorbed onto the adsorbent by taking the place of two Na(+) ions. The maximum exchangeable Na(+) present on the adsorbent was measured with the proton titration experiments and it was up to 1.0 mmol x Na(+)g(-1) dry adsorbent. To a significant extent, pH influenced the extraction of lead from aqueous solutions. The lead removal efficiency was up to 71%, 87% and 91% with initial solution pH at 3.0, 3.6 and 4.2, respectively. The Langmuir equation fitted the adsorption isotherm data well. The maximum adsorption capacity of lead calculated was 57.5, 76.9 and 114.9 mg lead g(-1) dry adsorbent at initial solution pH of 3.0, 3.6 and 4.2, respectively. Therefore, the adsorbent does offer favorable characteristics in lead removal from acidic wastewater. PMID- 12909110 TI - Time and space patterns of volatile organic compounds in a sewage treatment plant. AB - 47 regulated and non-regulated volatile organic compounds (VOCs) were characterised by closed-loop stripping analysis (CLSA) and high resolution gas chromatography coupled to mass spectrometry (HRGC/MS) in 28 aqueous samples from 4 sampling points along a sewage treatment plant in Manresa, Catalonia, Spain. A 4 x 2(2) factorial design (16 samples) was first prepared for the sampling, and reinforced with 12 additional samples at the plant influent. The total analyte weighted mean concentration was 232 microg x l(-1) at the plant influent, with a mass flow of 2231 kg x yr(-1). Petroleum solvents and terpenic compounds accounted for 79% of the influent analyte concentration. VOC concentration in influent was clearly higher for most VOCs from 12 to 22 h (high organic load hours), and lower from 24 to 10 h (lower organic load). Differences between time bands were confirmed through t tests. Differences between weekdays and the weekend were not so clear, and could not be confirmed through t tests. VOC concentrations along the plant are discussed. Overall analyte removal in the plant was 89%. PMID- 12909111 TI - Formulation of a mathematical model to predict solar water disinfection. AB - A mathematical model was formulated that will facilitate the prediction of solar disinfection by analyzing the effect of sunlight exposure (x(1)) and the load of bacterial contamination (x(2)), as predictor variables, on the efficiency of solar disinfection (y). Aliquots of 0.1 ml containing average numbers of E. coli, ranging between 1 and 5 x 10(3)cells/ml raw water, were introduced into each of the 96 wells of polystyrene microtitre plates. Plates, with the lid on, were exposed to sunlight for varying exposures ranging between 1.04 x 10(3) and 8.40 x 10(3)kJ m(-2). Double strength nutrient broth was then added. After 48 h incubation wells containing visible contamination were considered as containing one cell or more that survived the exposure. Data showed that disinfection is dependent both on the load of bacterial contamination and sunlight exposure. This relationship is characterized by curves having shoulders followed by a steep decline and then tailing off in an asymptotic fashion. The shoulder size increased with the increase of the contamination load, however, the slope remains the same. Statistical analysis indicates a positive correlation among the variables (R(2) = 0.893); the mathematical model, y=1-(1-e(-kx(1)))(x(2)), represents the relationship, with k being the solar inactivation constant. The exposure required to produce a given decontamination level can be predicted using the equation: x(1)=-1/kln[1-(1-y)(-1/x(2))]e(-micro/rho.m/A), where micro is the linear attenuation coefficient (m(-1)), rho is the density, m is the mass and A is the area of the exposed part of the sample. The predictor variables (x(1), x(2)) strongly influence the efficiency of solar disinfection, which can be predicted using the suggested mathematical model. The present data provides a means to predict the efficiency of solar disinfection as an approach to improve the quality of drinking water mainly in developing countries with adequate sunshine all year-round. PMID- 12909112 TI - Modeling of phosphorus dynamics in aquatic sediments: I--model development. AB - A model is developed to study the phosphorus dynamics in aquatic sediments and to conduct dynamic predictions of phosphorus release across a sediment-water interface. The model focuses on the sediment active layer below the sediment water interface and is based on primary mechanisms regulating phosphorus behavior in sediments, including effective diffusion, bioturbation mixing and burial processes (transport), organic decomposition, sorption kinetic processes and non linear partitioning (mobilization). The effects of environmental factors such as dissolved oxygen and temperature are taken into account. The model is solved by numerical integration. The primary difference from models in the literature is that the model directly describes the dynamic behavior of dissolved, particulate exchangeable ortho-phosphorus and organic phosphorus in sediments, and incorporates dynamic sorption and non-linear partitioning processes. These improve model mechanisms and allow regulation of phosphorus flux through the sediment reservoir that acts as both a source and sink of phosphorus. PMID- 12909113 TI - Modeling of phosphorus dynamics in aquatic sediments: II--examination of model performance. AB - A sediment phosphorus model, describing dynamics of organic phosphorus, dissolved reactive phosphorus and exchangeable particulate phosphorus, is applied to five monitoring stations in Chesapeake Bay, USA, to examine model performance in predicting sediment-water phosphorus exchange. The model was fit to 1 year of field measurements, and verified over 3 years at five sampling stations. The results show that the simulated concentrations of phosphorus reasonably correspond to model mechanisms and field observations in both spatial and temporal variations. Predicted release fluxes of phosphorus are consistent with field measurements and improved over those obtained by existing sediment phosphorus models. The model incorporates the effects of dissolved oxygen and non linear, non-equilibrium sediment sorption in phosphorus dynamics. A sensitivity analysis indicates the importance of the non-linear, non-equilibrium sorption processes to accurate prediction of sediment-water phosphorus flux. PMID- 12909114 TI - Removal of lead from contaminated water bodies using sea nodule as an adsorbent. AB - Adsorption of water soluble lead on polymetallic sea nodule has been studied in detail. Complete decontamination of lead is possible by appropriate sea nodule dosing. Adsorption is also dependent on pH and best adsorption is achieved at pH 6. Beyond pH 6, the desorption of lead from sea nodule surface is practically zero. Residual metal concentrations in the filtrate after adsorption is negligible. Both Freundlich and Langmuir isotherms may reasonably explain adsorption of lead on sea nodule. Chemically bound moisture plays a very crucial role in lead adsorption. Lead adsorptive capability of sea nodule is practically destroyed when calcined at a temperature of 900 degrees C. Lead loading capacity of sea nodule has been estimated at 440 mg of lead per gram of sea nodule. The performance of sea nodule as a lead adsorbent has been successfully tested over six simulated lead contaminated water systems. Lead loading capacity of sea nodule compares favorably with other adsorbents like activated carbon, ion exchange resin, anionic clay, granulated blast furnace slag and natural and treated zeolites. PMID- 12909115 TI - Development of multimetal binding model and application to binary metal biosorption onto peat biomass. AB - Biosorption of Cr(3+), Cu(2+) and Cd(2+) from binary metal solutions onto peat in the batch systems was investigated at pH 4. The order of maximum uptake was Cr>or=Cu>Cd and maximum uptake levels of ca. 0.4 mmol/g were observed for chromium and copper while cadmium was taken up to a maximum of ca. 0.2 mmol/g. Co ion competition resulted in up to 70 percent decrease of primary metal uptake. A novel approach to multicomponent sorption modelling involving regression to the total metal taken up was adopted. Two extended Langmuir-type models were found to exhibit good fit to the experimental data. Using the simpler model of these, three-dimensional sorption surfaces were generated which describe the metal uptake as a function of equilibrium concentrations of both metals. These methods allow prediction of metal uptakes over a continuum of concentrations of both metals in binary systems. PMID- 12909116 TI - Fecal indicator bacteria are abundant in wet sand at freshwater beaches. AB - Potential fecal contamination of sand in the wave-washed zone of public bathing beaches is overlooked in beach monitoring programs. Activity in this zone can bring pathogens to the sand surface or into the water, presenting a health risk to sensitive populations. On a unit weight basis (colony forming units per 100g), the mean summer abundance of the fecal indicator bacteria enterococci and Escherichia coli was 3-38 times higher in the top 20 cm of wet-sand cores than in the water column at six freshwater bathing beaches. E. coli were 4 times more abundant than enterococci in water but counts were similar in the sand. A correlation (r=0.60) existed between E. coli counts in the water and in the top 5 cm of sand only, whereas no relationship existed between enterococci abundance in water and sand. In general, enterococci were most numerous in the 5-10 cm sand stratum and E. coli in the 0-5 cm stratum. These preliminary data show that wet freshwater beach sand is a reservoir of fecal indicator bacteria. Enteric pathogens may also be present in beach sand. PMID- 12909117 TI - Cadmium and lead adsorption by a natural polysaccharide in MF membrane reactor: experimental analysis and modelling. AB - In the present work, Pb(2+) and Cd(2+) adsorption onto a natural polysaccharide has been studied in membrane reactors. The process involves a stirred semi-batch reactor for the adsorption step and a microfiltration (MF) process in order to confine the particles. Due to their lower affinity for the biosorbent, Cd(2+) ions were found to breakthrough the process faster than Pb(2+) cations. The experimental results showed the technical feasibility of the pilot. A mass balance model based on the Langmuir equilibrium isotherm was used to describe the adsorption process. This relation is able to predict experimental data under different operating conditions: the adsorbent and metal concentrations, and the permeate flow rate. Based on these results, it is demonstrated that the biosorbent studied represents an interesting low-cost solution for the treatment of metal ions polluted waters. PMID- 12909118 TI - Photocatalytic degradation of aqueous pollutants using silica-modified TiO(2). AB - Photocatalytic degradation (PCD) of several aqueous pollutants was investigated using a porous silica-coated titanium dioxide (SiO(2)-TiO(2)) photocatalyst. Several cationic, neutral and anionic pollutants were tested. The results indicate that modifying the surface properties of TiO(2) using silica significantly enhances the PCD rate of the cationic pollutants. The rate enhancement decreased with an increase in substrate concentration, especially for the quaternary amines, and was attributed to the decrease in initial adsorption. However, no significant rate-increase resulted for acetate and phenol. Results suggest that the increased presence of cationic pollutants at the catalyst surface caused the rate enhancement. PMID- 12909119 TI - The system design of atrazine oxidation by catalytic oxidation process through a kinetic approach. AB - A design method for the removal of atrazine (ATZ) by catalytic oxidation process (COP) was developed though examination of the reaction kinetics in various conditions. The reaction kinetics of ATZ decay in COP are found to consist of a reaction and a stagnant stage, while two characteristic constants, the initial decay rate and the oxidative capacity of the COP, are successfully derived and used to describe the two stages, respectively. The proposed model and the way to solve the two characteristic constants (the initial decay rate and the oxidative capacity) are capable of describing the ATZ degradation precisely in COP at different Fe(II) concentrations and [Fe(II)]/[H(2)O(2)] ratios, where the marginal [Fe(II)]/[ATZ] ratio at 10 is found to be the critical point to achieve a cost-effective treatment of ATZ by COP. A series of equations for practical completely mixed batch reactor design is also developed based on the two characteristic constants. PMID- 12909120 TI - Adsorption of halogenated aromatic pollutants by a protein released from Bacillus pumilus. AB - Previous studies of the biosorption of halogenated aromatic pollutants (HAPs) have focused on the sorption of these compounds by cell bodies. However, in this study we investigated the adsorption of HAPs by biocompounds released from a bacterium, Bacillus pumilus. When B. pumilus was exposed to high temperature, it released a protein and carbohydrates, exclusively. After determining experimental conditions using 1,2,3,4-tetrachlorinated dibenzofuran (1,2,3,4-TCDF), the adsorption characteristics of polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs), polychlorinated biphenyls (PCBs), chlorinated benzenes, and chlorinated naphthalenes were investigated. These HAPs were adsorbed considerably not only by cells but also by the released protein. In general, highly chlorinated congeners were adsorbed to a greater extent on the protein than lowly chlorinated ones, and the amount adsorbed differed between isomers. The present results are consistent with adsorption occurring via a passive physico-chemical mechanism. Finally, the importance of biocompounds released or excreted from microorganisms for the removal of HAPs is discussed. PMID- 12909121 TI - Quantification of denitrification potential in carbonaceous trickling filters. AB - Biofilm samples from a carbonaceous trickling filter (TF) were evaluated in bench scale reactors to determine their maximum potential denitrification rates. Intact, undisturbed biofilms were placed into 0.6 L bench-scale reactors filled with sterilized, primary clarifier effluent spiked with nitrate to a final concentration of 16-18 mg/L as N. Dissolved oxygen concentrations were maintained between 2 and 4 mg/L in the bulk aqueous phase. Nitrate loss from the reactors was monitored over a 5h period. Denitrification rates of 3.09-5.55 g-N/m(2)day were observed with no initial lag period. This suggests that the capacity for denitrification is inherent in the biofilm and that denitrification can take place even when oxygen is present in the bulk aqueous phase. There were no significant differences in denitrification rates per unit area of media (g N/m(2)day) either between (a). experimental runs or (b). sampling locations over the trickling filter. This suggests that denitrification potentials are uniform over the entire volume of the full-scale TF. For wastewater treatment plants with TFs that currently nitrify downstream, this approach may be used to meet less stringent permitted discharge concentrations and may allow some facilities to postpone or eliminate construction of additional unit processes for denitrification. PMID- 12909122 TI - Hybrid flotation--membrane filtration process for the removal of heavy metal ions from wastewater. AB - A promising process for the removal of heavy metal ions from aqueous solutions involves bonding the metals firstly to a special bonding agent and then separating the loaded bonding agents from the wastewater stream by separation processes. For the separation stage, a new hybrid process of flotation and membrane separation has been developed in this work by integrating specially designed submerged microfiltration modules directly into a flotation reactor. This made it possible to combine the advantages of both flotation and membrane separation while overcoming the limitations. The feasibility of this hybrid process was proven using powdered synthetic zeolites as bonding agents. Stable fluxes of up to 80l m(-2)h(-1) were achieved with the ceramic flat-sheet multi channel membranes applied at low transmembrane pressure (<100 mbar). The process was applied in lab-scale to treat wastewater from the electronics industry. All toxic metals in question, namely copper, nickel and zinc, were reduced from initial concentrations of 474, 3.3 and 167mg x l(-1), respectively, to below 0.05 mg x l(-1), consistently meeting the discharge limits. PMID- 12909123 TI - Storm impacts upon the composition of organic matrices in Nagara River--a study based on molecular weight and activated carbon adsorbability. AB - The impacts of a heavy storm of rain on the composition of natural organic matter (NOM) in Nagara River water were studied in terms of molecular weights (MWs) and activated carbon (AC) adsorbabilities using six water samples collected during a critical Typhoon weather condition. The composition in MWs was analyzed using a HPSEC system and that in adsorbabilities was characterized using parameters devised to reflect NOMs average adsorptive strength (K(M)), adsorptive strength polydispersity (sigma), affinity to AC (1/n) and non-adsorbable fraction (C(non)/C(T0)), respectively. These parameters were determined by model description of observed isotherms with a distributed fictive component method. The heavy storm of rain brought higher content of larger organic components into the river source, thus causing changes of NOMs weight-averaged MWs in the range of 2962-3495 Dalton and MW polydispersity in the narrow range of 1.153-1.226. Comparison of K(M) and sigma values for all samples assessed with both indices of TOC and UV260 showed that large proportions of the storm-induced organic components had adsorptive strengths similar to those existent before the storm, with the presence levels for components revealing much strong and weak adsorbabilities being low. Among all organic components brought into the river by the storm of rain, the percentages of non-adsorbable ones was lower (smaller C(non)/C(T0) values); and the adsorbable ones had generally more affinity to the adsorbents used (smaller 1/n values). PMID- 12909124 TI - Removal of cadmium and nickel from wastewater using bagasse fly ash--a sugar industry waste. AB - The bagasse fly ash, an industrial solid waste of sugar industry, was used for the removal of cadmium and nickel from wastewater. As much as 90% removal of cadmium and nickel is possible in about 60 and 80 min, respectively, under the batch test conditions. Effect of various operating variables, viz., solution pH, adsorbent dose, adsorbate concentration, temperature, particle size, etc., on the removal of cadmium and nickel has been studied. Maximum adsorption of cadmium and nickel occurred at a concentration of 14 and 12 mg x l(-1) and at a pH value of 6.0 and 6.5, respectively. A dose of 10 g x l(-1) of adsorbent was sufficient for the optimum removal of both the metal ions. The material exhibits good adsorption capacity and the adsorption data follow the Langmuir model better then the Freundlich model. The adsorption of both the metal ions increased with increasing temperature indicating endothermic nature of the adsorption process. Isotherms have been used to determine thermodynamic parameters of the process, viz., free energy change, enthalpy change and entropy change. PMID- 12909125 TI - The simultaneous removal of calcium and chloride ions from calcium chloride solution using magnesium-aluminum oxide. AB - We investigated the removal of Ca(2+) and Cl(-) from CaCl(2) solution at 20-60 degrees C, using magnesium-aluminum oxide, Mg(0.80)Al(0.20)O(1.10), prepared by the thermal decomposition of a hydrotalcite-like compound, Mg(0.80)Al(0.20)(OH)(2)(CO(3))(0.10).0.78 H(2)O. The degree of Ca(2+) and Cl(-) removal from the solution increased with increasing initial CaCl(2) concentration, temperature, and quantity of Mg(0.80)Al(0.20)O(1.10) added. When Mg(0.80)Al(0.20)O(1.10) was added to 0.25 M CaCl(2) solution in a Mg(0.80)Al(0.20)O(1.10)/CaCl(2) molar ratio of 20, the degree of Ca(2+) and Cl(-) removal from the solution at 60 degrees C after 0.5 h was 93.0% and 98.2%, respectively. These results reveal that Mg(0.80)Al(0.20)O(1.10) has the capacity to remove Ca(2+) and Cl(-) simultaneously from aqueous solution. PMID- 12909126 TI - The human antibody repertoire to infectious agents: implications for disease pathogenesis. AB - Antibodies are critical entities used for the protection of an individual organism against infection and disease. Through a complex series of events genes rearrange to encode repertoires of molecules, which are tested for their ability to identify foreign molecules. In this report, we discuss factors defined by the individual as well as by infectious organisms that shape the antibody repertoire. A more thorough understanding of the interplay between these factors will eventually allow us to elucidate the underlying mechanisms of disease susceptibility following infection, and to design potent vaccines and other immunomodulating reagents. PMID- 12909127 TI - Membrane-bound complement regulatory proteins inhibit complement activation by an immunotherapeutic mAb in a syngeneic rat colorectal cancer model. AB - MAb-mediated immunotherapy offers a potential tool for destroying metastasizing colorectal tumor cells. Promising results have been obtained by using xenograft models. However, overexpression of membrane-bound complement regulatory proteins (mCRP) impedes complement-mediated destruction of tumor cells in vitro. mCRP operate in a species selective manner. Therefore a syngeneic animal model is needed to investigate the contribution of mCRP in mAb-mediated immunotherapy. Here we present a syngeneic rat colorectal carcinoma model, which fulfills the conditions necessary to investigate the effect of mCRP expression on mAb-mediated immunotherapy of metastases of solid tumors.CC531 rat colorectal cancer cells were injected subcapsularly into the liver of syngeneic WAG/Rij rats. Four mAb (MG1(IgG2a), MG2(IgG2a), MG3(IgG3) and MG4(2a)(IgG2a)) directed against CC531 cells, were tested for their complement activating abilities in vitro and tumor homing capacities in vivo. Only MG4(2a) was found to activate complement in vitro and home to the tumor cells in vivo. This mAb induced C3-deposition and C mediated lysis of CC531 cells in vitro when the effects of the C-inhibitors Crry/p65 and CD59 were neutralized. This implies an important role for these mCRP in restricting the effector functions of tumor-associated mAb on these cells. Although C activation could be induced by MG4(2a) in situ on tumor tissue sections, no deposition of C3 could be found on the tumor cells positive for MG4(2a) in vivo. This suggests that complement activation in vivo was inhibited by mCRP. The results indicate the suitability of this syngeneic animal model for studying the effects of mAb immunotherapy. However, the effect of mCRP on tumor cells need to be overcome, e.g. by the use of mAb against tumor antigens and mCRP. PMID- 12909128 TI - Selection and characterization of naturally occurring single-domain (IgNAR) antibody fragments from immunized sharks by phage display. AB - The novel immunoglobulin isotype novel antigen receptor (IgNAR) is found in cartilaginous fish and is composed of a heavy-chain homodimer that does not associate with light chains. The variable regions of IgNAR function as independent domains similar to those found in the heavy-chain immunoglobulins of Camelids. Here, we describe the successful cloning and generation of a phage displayed, single-domain library based upon the variable domain of IgNAR. Selection of such a library generated from nurse sharks (Ginglymostoma cirratum) immunized with the model antigen hen egg-white lysozyme (HEL) enabled the successful isolation of intact antigen-specific binders matured in vivo. The selected variable domains were shown to be functionally expressed in Escherichia coli, extremely stable, and bind to antigen specifically with an affinity in the nanomolar range. This approach can therefore be considered as an alternative route for the isolation of minimal antigen-binding fragments with favorable characteristics. PMID- 12909129 TI - A set of genes selectively expressed in murine dendritic cells: utility of related cis-acting sequences for lentiviral gene transfer. AB - Professional antigen presenting cells such as dendritic cells (DC) and macrophages (Mphi) share similar characteristics; however, they differ in their ability to initiate an immune response. DCs are much more potent in priming and stimulating nai;ve T-cells. Thus, DCs are good targets for the expression of foreign genes to elicit and specifically modify immune responses. To identify DC markers cDNA subtraction was performed using murine MHC class II(high), B7(high) bone marrow derived DCs as tester and interferon-gamma/E. coli lipopolysaccaride (LPS) treated bone marrow derived macrophages as driver. Analysis of 114 resulting clones revealed a diverse pattern of DC selective (DC(DeltaMphi)) gene expression including known genes whose expression in DCs had not been previously demonstrated as well as multiple novel genes. For several identified DC(DeltaMphi) genes, proximal promoter elements were isolated and incorporated into self-inactivating lentiviral GFP reporter vectors. Promoter activity was measured in bone marrow derived macrophages or dendritic cells. Of the promoters analyzed those for B7-DC and CCL17 drove strong GFP expression in DCs but not in resting or activated macrophages. The CCL17 promoter offered the highest level of expression in DCs and was further activated by culture with LPS or interleukin-4 (IL-4). In contrast, the B7-DC promoter was induced by IL-4 but not by LPS. Endogenous CCL17 and B7-DC mRNAs were increased similarly in IL-4 cultured DCs but only CCL17 was induced by LPS. Additionally, IL-4 increased cell surface expression of B7-DC in both immature and mature DCs. PMID- 12909130 TI - Relative importance of different human aPL derived heavy and light chains in the binding of aPL to cardiolipin. AB - INTRODUCTION: Previous studies have suggested the importance of somatic mutations and arginine, asparagine and lysine residues in the complementarity determining regions (CDRs) of antiphospholipid antibodies (aPL) implicated in the pathogenesis of the antiphospholipid antibody syndrome. The relative contributions of the heavy and light chains of aPL in binding to cardiolipin (CL) were assessed by pairing the heavy and light chains of two IgG, beta(2)GPI dependent aPL (IS4 and CL24) with different partner chains from other IgG, beta(2)GPI independent aPL (UK4) and anti-DNA antibodies (B3 and 33H11). METHODS: Four heavy (V(H)) and five light (V(L)) chain variable sequences from three aPL and two anti-DNA antibodies were cloned into expression vectors containing appropriate gamma(1), lambda or kappa constant region cDNA. Paired combinations of heavy and light chain expression plasmids were transfected into COS-7 cells allowing transient expression of whole IgG molecules, which were harvested and tested for the ability to bind CL and DNA by enzyme-linked immunosorbant assay (ELISA). RESULTS: Whole IgG was produced from 19 heavy/light chain combinations. IS4V(H) was dominant in conferring the ability to bind CL with four of the five V(L) tested. The identity of the V(L) region paired with IS4V(H) was important in determining the strength of binding to CL. IS4V(H) contains multiple arginine residues in CDR3, which may have accumulated due to antigen driven selection. It is likely that these arginine residues may interact with CL. The combination B3V(H)/B3V(L) also bound CL, but none of the other 14 combinations showed any binding in this assay. CONCLUSION: Whole IgG molecules capable of binding CL were produced by in vitro expression in COS-7 cells. Arginine residues play important roles in binding to CL and double-stranded DNA. However, different patterns of mutation to arginine are associated with binding to each of these antigens. PMID- 12909131 TI - Unusually long germline DH genes contribute to large sized CDR3H in bovine antibodies. AB - We demonstrated earlier the existence of an exceptionally long third complementarity-determining region of the heavy chain (CDR3H) (up to 61 amino acids (aa)), with multiple cysteine residues, in some functional IgM antibodies of cattle. To understand the origin of such a long CDR3H, we have now characterized the germline diversity gene (D(H)) of the cattle. A 2.3kb genomic DNA fragment hybridizing with a newly developed DNA probe to putative bovine D(H) gene sequences was isolated, cloned and its nucleotide sequence determined. Inspection of the nucleotide sequence led to identification of three bovine germline D(H) gene segments of varying size: 42bp (14 possible codons), 58bp (19 possible codons) and 148bp (49 possible codons). The characteristic repetitive GGT and TAT codons, remarkable in the CDR3H region of fetal VDJ rearrangements likely encoded by germline genes, are noted in two of the identified germline D(H) genes. These D(H) genes are preferentially expressed in the third reading frame to encode hydrophilic glycine and tyrosine residues in the CDR3H region. Phylogenetic analysis suggests that bovine D(H) genes are closest to rabbit and chicken D(H) genes. Thus, both short and long germline D(H) genes exist in cattle and these are capable of directly contributing to CDR3H size heterogeneity including the exceptionally long CDR3H region, apart from recombination associated mechanistic factors. PMID- 12909133 TI - Semi-quantitative analysis of indigo carmine, using silver colloids, by surface enhanced resonance Raman spectroscopy (SERRS). AB - The application of surface enhanced resonance Raman spectroscopy (SERRS) to the semi-quantitative analysis of the dye, indigo carmine, has been examined using citrate-reduced silver colloids. Good linear correlations are observed for the dye band at 1580 cm(-1) in the concentration range 10(-7)-10(-5) and 10(-9)-10( 5) mol dm(-3), using laser exciting wavelengths of 514.5 [(R=0.9983)] and 632.8 nm [(R=0.9978)], respectively. At concentrations of dye above 10(-6) M the concentration dependence of the SERRS signals is non-linear due to the coverage of the surface of the colloidal particles by the dye being in excess of a full monolayer. At concentrations above 10(-6) M resonance Raman spectroscopy (RRS) can be employed for the quantitative analysis of the dye. An internal standard was used and a good linear correlation (R=0.997) was observed for the dependence of dye signal intensities at 1580 cm(-1) in the concentration range 10(-5)-10(-4) M using a laser exciting wavelength of 514.5 nm. The limits of detection of indigo carmine by SERRS (514.5 nm), SERRS (632.8 nm) and solution RRS (514.5 nm) are found to be 0.9, 1 and 38 ppm, respectively. PMID- 12909134 TI - Raman spectra of carotenoids in natural products. AB - Resonance Raman spectra of naturally occurring carotenoids have been obtained from nautilus, periwinkle (Littorina littorea) and clam shells under 514.5 nm excitation and these spectra are compared with the resonance Raman spectra obtained in situ from tomatoes, carrots, red peppers and saffron. The tomatoes, carrots and red peppers gave rise to resonance Raman spectra exhibiting a nu1 band at ca. 1520 cm(-1), in keeping with its assignment to carotenoids with ca. nine conjugated carbon-carbon double bonds in their main chains, whereas the resonance Raman spectrum of saffron showed a nu1 band at 1537 cm(-1) which can be assigned to crocetin, having seven conjugated carbon-carbon double bonds. A correlation between nu1 wavenumber location and effective conjugated chain length has been used to interpret the data obtained from the shells, and the wavenumber position (1522 cm(-1)) of the nu1 band of the carotenoid in the orange clam shell suggests that it contains nine conjugated double bonds in the main chain. However, the black periwinkle and nautilus shells exhibit nu1 bands at 1504 and 1496 cm(-1), respectively. On the basis of the correlation between nu1 wavenumber location and effective conjugated chain length, this indicates that they contain carotenoids with longer conjugated chains, the former having ca. 11 double bonds and the latter ca. 13 or even more. Raman spectra of the nautilus, periwinkle and clam shells also exhibited a strong band at 1085 cm(-1) and a doublet with components at 701 and 705 cm(-1), which can be assigned to biogenic calcium carbonate in the aragonite crystallographic form. PMID- 12909135 TI - Semi-quantitative analysis of indigo by surface enhanced resonance Raman spectroscopy (SERRS) using silver colloids. AB - In this paper we report for the first time semi-quantitative analysis of indigo using surface enhanced Raman spectroscopy (SERS) and surface enhance resonance Raman spectroscopy (SERRS). Indigo, a dye widely used today in the textile industry, has been used, historically, both as a dye and as a pigment; the latter in both paintings and in printed material. The molecule is uncharged and largely insoluble in most solvents. The application of SERS/SERRS to the semi quantitative analysis of indigo has been examined using aggregated citrate reduced silver colloids with appropriate modifications to experimental protocols to both obtain and maximise SERRS signal intensities. Good linear correlations are observed for the dependence of the intensities of the SERRS band at 1151 cm( 1) using laser exciting wavelengths of 514.5 nm (R=0.9985) and 632.8 nm (R=0.9963) on the indigo concentration over the range 10(-7)-10(-5) and 10(-8) 10(-5) mol dm(-3), respectively. Band intensities were normalised against an internal standard (silver sol band at 243 cm(-1)). Resonance Raman spectra (RRS) of aqueous solutions of indigo could not be collected because of its low solubility and the presence of strong fluorescence. It was, however, possible to obtain RS and RRS spectra of the solid at each laser excitation wavelength. The limits of detection (L.O.D.) of indigo by SERS and SERRS using 514.5 and 632.8 nm were 9 ppm at both exciting wavelengths. Signal enhancement by SERS and SERRS was highly pH dependent due to the formation of singly protonated and possibly doubly protonated forms of the molecule at acidic pH. The SERS and SERRS data provide evidence to suggest that an excess of monolayer coverage of the dye at the surface of silver colloids is observed at concentrations greater than 7.85x10(-6) mol dm(-3) for each exciting wavelength. The data reported herein also strongly suggest the presence of multiple species of the indigo molecule. PMID- 12909136 TI - Raman spectroscopy of different types of Mexican copal resins. AB - Dispersive Raman spectra of six copal resins, that were purchased in local markets in Mexico, are presented. The spectra were interpreted and compared with each other. For all these spectra, the relative intensity of the Raman band at approximately 1645 cm(-1), attributed to the exomethylene nu(C=C) stretching vibration, was rather low, especially as fresh samples are involved. In one resin, viz. Incienso, CaCO3 was detected. Probably this inorganic pigment was added as a whitener. In the spectrum of Lagrima a starch fraction was present. Raman spectra of a sample from an Aztec figurine were recorded. It was shown that its composition was inhomogeneous at the micrometer level. Here, too, CaCO3 was observed. It was not possible to identify the resin applied in the antique figurine due to material degradation by age and environmental exposure. PMID- 12909137 TI - Vibrational spectra of the hydrated carbonate minerals ikaite, monohydrocalcite, lansfordite and nesquehonite. AB - The Raman (200-4000 cm(-1)) and infrared (600-4000 cm(-1)) spectra of four rare carbonate hydrate minerals are reported. These are naturally occurring and synthetic ikaite CaCO3.6H2O, and nesquehonite MgCO3.3H2O; natural monohydrocalcite CaCO3.H2O, and synthetic lansfordite MgCO3.5H2O. The spectra of synthetic ikaite partially substituted with 2H2O and also with 13C were measured, as were those of synthetic deuteriated nesquehonite. Spectra of ikaite and lansfordite, both of which decompose at room temperatures, were measured below 0 degrees C. Assignments of fundamental modes are proposed. PMID- 12909138 TI - Raman spectroscopy of selected arsenates--implications for soil remediation. AB - The contamination of soils with heavy metals such as As, Cr and Cu is of great importance; the remediation of such soils even more so. Arsenic compounds are prevalent in soils either through leaching of mine tailings, the use of Cu/Cr/As as a wood preservative or through the use of arsenic in cattle dips. The arsenic compounds in soils and leachates can be highly reactive and mobile, resulting in the formation of metal arsenate compounds. Of these compounds, one such set of minerals that can be formed is the vivianite arsenate minerals. Raman spectroscopy has been used to characterise the vivianite arsenates and to identify arsenic contaminants in a soil. PMID- 12909139 TI - Catalogue of 45 reference Raman spectra of minerals concerning research in art history or archaeology, especially on corroded metals and coloured glass. AB - Small catalogues of reference Raman spectra of interest for analysing geomaterials or biomaterials of relevance to art history or archaeology are gradually being published by different research groups. However, except for some older catalogues, they are all concerned primarily with pigments, whether inorganic or organic. Here we present for the first time a catalogue of Raman spectra of minerals that may be found in corroded metal artworks or artefacts. At the same time we include some inorganic pigments that may be found in or on stained glass. Most of the minerals analysed came from the Gallery of Mineralogy at the Museum National d'Histoire Naturelle and most were verified by X-ray diffraction in order to augment the confidence in the mineral identity (which is not the case with many other catalogues). A number of problems encountered with mineral terminology are discussed. Comments are made on the spectra where appropriate. PMID- 12909140 TI - On the characterization of disordered and heterogeneous carbonaceous materials by Raman spectroscopy. AB - The applicability of Raman spectroscopy to characterize disordered and heterogeneous carbonaceous materials (CM) is discussed, by considering both natural and synthetic CM. First, different analytical mismatches during the measurement are discussed and technical indications are provided in order to eliminate them. Second, the accuracy and relevance of the different parameters obtained by the decomposition of spectra by conventional fitting procedure, is reviewed. Lastly, a new Raman technique (Raman area mode microspectroscopy) giving an homogeneous repartition of power within a large laser beam is presented, this technique being powerful to study strongly heterogeneous CM and/or photosensitive samples. PMID- 12909141 TI - Raman spectroscopic detection of biomolecular markers from Antarctic materials: evaluation for putative Martian habitats. AB - The vital UV-protective and photosynthetic pigments of cyanobacteria and lichens (microbial symbioses) that dominate primary production in Antarctic desert ecosystems auto-fluoresce at short-wavelengths. A long wavelength (1064 nm) near infra-red laser has been used for non-intrusive Raman spectroscopic analysis of their ecologically significant compounds. There is now much interest in the construction of portable Raman systems for the analysis of cyanobacterial and lichen communities in the field; to this extent, Raman spectra obtained with laboratory-based systems operating at wavelengths of 852 and 1064 nm have been evaluated for potential fieldwork applications of miniaturised units. Selected test specimens of the cyanobacterial Nostoc commune, epilithic lichens Acarospora chlorophana, Xanthoria elegans and Caloplaca saxicola and the endolithic Chroococcidiopsis from Antarctic sites have been examined in the present study. Although some organisms gave useable Raman spectra with short-wavelength lasers, 1064 nm was the only excitation that was consistently excellent for all organisms. We conclude that a 1064 nm Raman spectrometer, miniaturised using an InGaAs detector, is the optimal instrument for in situ studies of pigmented communities at the limits of life on Earth. This has practical potential for the quest for biomolecules residual from any former surface or subsurface life on Mars. PMID- 12909142 TI - Lead-tin mirror formation from mixtures of red lead and tin sulphide. AB - Mosaic gold, tin (IV) sulphide, is a yellow pigment which was known in antiquity but whose use was superseded by other more easily obtainable yellow pigments by the Renaissance. The identification of mosaic gold residues in a burnished golden mirror decoration on a XIIIth Century Spanish polychrome statue is important in that the first reference to the use of mosaic gold in the European literature dates from the XIVth Century, although the use of this material in China had been recorded some time before. In this paper, Raman spectroscopy, XRD and SEM are used in the analysis of the conditions required for the formation of golden mirrors using tin (IV) sulphide in admixture with dilead (II) lead (IV) tetroxide and mercury (II) sulphide. From these results, it is proposed that the major reactions are the reduction of Sn(IV) to Sn(0) with the accompanying oxidation of lead (II) oxide to lead (IV) oxide and the formation of lead (0) and lead (II) sulphide. From these results it was possible to explain the process of creation of the golden mirror from mosaic gold in the XIIIth Century. PMID- 12909143 TI - Non-destructive analysis of pigments and other organic compounds in lichens using Fourier-transform Raman spectroscopy: a study of Antarctic epilithic lichens. AB - Lichens in Antarctic habitats are subjected to environmental extremes, including UVB radiation, desiccation and low temperatures, as well as to rapid fluctuations in these. Lichens synthesise a variety of chemical compounds in response to their environmental conditions which contribute towards their colour, and which act as protectants against physiological stresses. The fluorescence generated by the lichens at 532 nm can be used in epifluorescence microscopy to identify their presence on substrata but this can severely affect the Raman spectra using visible excitation. The advantage of the near infrared excitation used in FT Raman spectroscopy in minimising fluorescence emission facilitates the molecular characterisation of lichen encrustations without having to remove the thallus from its substrate or remove or otherwise damage any part of the thallus. Spectroscopic biomarkers are proposed which allow the lichens to be characterised by the identification of characteristic lichen substances; the use of these biomarkers for the preliminary taxonomic identification of Antarctic lichens is examined and some potential pitfalls are described. PMID- 12909144 TI - Vibrational spectroscopic study of the contents of a chest excavated from the wreck of the HMS Pandora. AB - The FT-IR and Raman spectroscopic analysis of a red powder found in a chest from an officer's cabin during the excavation of the wreck of the 18th Century frigate HMS Pandora have confirmed that the pigment is cinnabar, mercury(II) sulphide. Weaker signals in the Raman spectrum are assignable to a proteinaceous material, such as collagen, typical of a degraded vellum or parchment. Comparison of the Raman spectra with that of a pigmented seal from a 1786 Lieutenant's commission demonstrated that the beeswax component of the seal was not observable. PMID- 12909145 TI - Characterization and surface-enhanced Raman spectral probing of silver hydrosols prepared by two-wavelength laser ablation and fragmentation. AB - A four step Ag foil laser ablation-Ag nanoparticle fragmentation procedure in ultrapure water was carried out both under argon and in air. Pulses of a high power Nd/YAG laser were used for laser ablation (1064 nm) and for the three step Ag hydrosol treatment in the absence of Ag foil in the sequence 1064-532-1064 nm. Transmission electron microscopy (TEM) and surface plasmon (SP) extinction spectra provide evidence of Ag nanoparticle fragmentation in the second and third step of the procedure carried out under argon. While polydispersity of Ag hydrosol increases in the second step, both the polydispersity and the mean size of the nanoparticles are reduced in the third step. Qualitative and quantitative surface-enhanced Raman scattering (SERS)/surface-enhanced resonance Raman scattering (SERRS) spectral probing of systems with Ag hydrosols and the selected adsorbates at 514.5 nm excitation shows that Ag hydrosols obtained in the second step of the preparation procedure carried out in air are the most suitable substrates for SERS/SERRS experiments performed at this excitation wavelength. PMID- 12909146 TI - Influence of laminar flow on preorientation of coal tar pitch structural units: Raman microspectroscopic study. AB - In order to estimate the role of laminar flow of viscous, aromatic matter of carbonaceous precursor on microtextural preorientation in pregraphitization stage, we performed experiments with coal tar pitch (CTP). The principal hypothesis of preorientation of basic structural units (BSUs) in the case of laminar flow (pressure impregnation of CTP into porous matrix) and secondary release of volatiles during carbonization were studied. Glass microplates, planar porous medium with average distance between single microplates 5 microm were used as suitable porous matrix. Samples of CTP were carbonized up to 2500 degrees C. Optical microscopy reveals large flow domains in the sample of cokes carbonized between glass microplates. Raman microspectroscopy and high resolution transmission electron microscopy (HRTEM) show that at nanometric scale, the samples do not support the proposed hypotheses. With increasing temperature of pyrolysis, the graphitization of CTP impregnated into porous matrix proceeds to lower degree of structural ordering in comparison with single pyrolyzed CTP. This is explained by the release of volatile matter during carbonization in geometrically restricted spaces. More evident structural changes were discovered with the sample of single coke, where parts of fine grain mosaics, relicts of 'so called QI parts', reveal higher structural organization, in comparison with large and prolonged flow domains, similar to flow domains of cokes from microplates. PMID- 12909147 TI - Raman spectroscopy of carbon and solid bitumens in sedimentary and metamorphic rocks. AB - Different types of carbonaceous matter from rocks display Raman spectral features which knowledge permits to obtain structural information of these materials. Application of Raman microspectroscopy to investigate kerogen, bitumen, fossils, highly carbonified amorphous carbon as well as graphite from different environments is reviewed. Differences in Raman spectra and structural differences between carbonaceous samples differing in their metamorphic history are discussed on the basis of new data. PMID- 12909148 TI - In situ mobile subaquatic archaeometry evaluated by non-destructive Raman microscopy of gemstones lying under impure waters. AB - A series of laboratory simulations have been made in order to evaluate the credibility of carrying out physico-chemical analysis of cultural heritage items by Raman spectral fingerprinting using a mobile Raman microscope in situ under natural impure water in subaquatic or submarine conditions. Three different kinds of gemstone (zircon, microcline and sodalite) were successively placed under different kinds of impure water into which a low power microscope objective was immersed to eliminate the normal aerial pathway between the objective and the object to be analysed. According to the nature of the impurities (inorganic or organic, dissolved or suspended, transparent or coloured) the results obtained variously gave Raman band intensities stronger than, similar to or weaker than those of spectra obtained without water, i.e. in air. The significant point is that after only minor spectral treatment the less good spectra nevertheless yielded exploitable data with most, if not all, of the key Raman bands being detected. Thus the problems of fluorescence or peak absences under water are of a similar degree of magnitude to the other problems inherent with the Raman spectroscopic technique in aerial conditions, e.g. relative peak intensities varying with crystal orientation; peak positions varying with chemical composition. These results indicate that even if at certain sites of submerged cities or sunken ships, the combination of animal, vegetal, mineral and microbial impurities join together to inhibit or hinder the success of subaquatic or submarine archaeometry, there will certainly be other sites where such activity is indeed credible. PMID- 12909149 TI - Raman microscopy of Greek icons: identification of unusual pigments. AB - Five Greek icons, made between the 15th and 18th centuries and now belonging to the Victoria and Albert Museum collections, were analysed by energy-dispersive X ray fluorescence (EDXRF), optical microscopy and Raman microscopy in order to determine the stratigraphy of the artworks and the identity of the pigments used. Together with common pigments, such as red lake, vermilion, red lead, red iron oxide, orpiment, yellow ochre, lead white, chalk, gypsum, anhydrite, Prussian blue, indigo and a copper-containing green, a few unusual materials were identified, specifically pararealgar (a yellow arsenic sulphide, As4S4), its precursor the chi-phase, and lead tin yellow type II (PbSn(1-x)SixO3). Attention is drawn to the complementarity of the techniques used for the pigment identifications. PMID- 12909150 TI - Stand-off Raman spectroscopic detection of minerals on planetary surfaces. AB - We have designed and developed two breadboard versions of stand-off Raman spectroscopic systems for landers based on a 5-in. Maksutov-Cassegrain telescope and a small (4-in. diameter) Newtonian telescope receiver. These systems are capable of measuring the Raman spectra of minerals located at a distance of 4.5 66 m from the telescope. Both continuous wave (CW) Ar-ion and frequency doubled Nd:YAG (532 nm) pulsed (20 Hz) lasers are used as excitation sources for measuring remote Raman spectra of rocks and minerals. We have also made complementary measurements on the same rock samples with a micro-Raman system in 180 and 135 degrees geometry for evaluating the system performance and for estimating effect of grain size and laser-induced heating on the spectra of minerals using alpha-quartz as a model mineral. A field portable remote pulsed Raman spectroscopic system based on the 5-in. telescope and an f/2.2 spectrograph has been developed and tested. We have also demonstrated a prototype of a combined Raman and laser-induced breakdown spectroscopy (LIBS) system, capable of providing major element composition and mineralogical information on both biogenic and inorganic minerals at a distance of 10 m from the receiver. PMID- 12909151 TI - Raman microspectrometric investigation of wall paintings in S. Giovanni Evangelista Abbey in Parma: a comparison between two artists of the 16th century. AB - Micro-Raman spectroscopy, combined with gas chromatography and ultra-violet fluorescence photography, was used to study some wall paintings in the S. Giovanni Evangelista Abbey in Parma, Italy. The restoration of some painted chapels enabled a comparison between two painters of the 16th century: Parmigianino (Girolamo Francesco Maria Mazzola, 1503-1540) and Michelangelo Anselmi (1492?-1556?). Micro-Raman spectroscopy determined the palette used by the artists, leading to the identification of different white, yellow, red, brown, green, blue and black pigments. Some pigments are evidence of later restorations. Gas chromatography combined with mass spectroscopy revealed the presence of organic binding media and enabled to distinguish between fresco and secco paintings. PMID- 12909152 TI - Clinical utility of an electronic poisons information and clinical decision support tool. AB - The objective of the study was to assess the use of a computer toxicology database/clinical decision aid by clinical practitioners. The study investigated the sources that Emergency Department (ED) personnel use to obtain toxicology information and performed a quality audit of the current database. A questionnaire survey of ED staff was used in departments with access to the New Zealand Poisons Centre Substance Database (NZSD), a toxicology CD ROM computer database. Outcome measures were reported use of alternative data sources when managing clinical toxicology presentations and the qualities of the NZSD. Computer databases are commonly used for the management of clinical toxicology cases and the toxicology computer database/clinical decision aid studied is well accepted and used in Emergency Medicine practice. The users of the NZSD assessed the usability and quality of the information of the database. PMID- 12909153 TI - Knowledge management in clinical practice: a systematic review of information seeking behavior in physicians. AB - OBJECTIVES: To determine information seeking behavior of physicians. DATA SOURCES: Systematic review of 19 studies that described information seeking behavior in a number of different settings using differing methodologies. Analysis was limited to quantitative studies describing sources of information sought by physicians. RESULTS: Investigators have used questionnaires, interviews and observation to identify the information seeking behavior of clinicians. The results were mainly obtained from trials in the United States and showed a wide variation in primary information sources used by physicians. The most frequent source for information used are text sources, second is asking colleagues and only one study found electronic databases to be the primary resource. Physician's desk reference is the commonest cited printed resource. Convenience of access, habit, reliability, high quality, speed of use, and applicability makes information seeking likely to be successful and to occur. The lack of time to search, the huge amount of material, forgetfulness, the belief that there is likely to be no answer, and the lack of urgency all hinder the process of answering questions. CONCLUSIONS: The wide variation in information seeking behavior implies a need for further categorization of information need and information sources. Careful planning of information delivery to physicians is required to enable them to keep up to date and to improve knowledge transfer. PMID- 12909154 TI - Preserving confidentiality when sharing medical database with the Cellsecu system. AB - We propose a computer system called Cellsecu that maintains the anonymity and the confidentiality of each cell containing sensitive information in medical database. Cellsecu attains this by automatically removing, generalizing, and expanding information. It is designed to enhance data privacy protection so a data warehouse can automatically handle queries. In most cases, health organizations collect medical data with explicit identifiers, such as name, address and phone number. Simply removing all explicit identifiers prior to release of the data is not enough to preserve the data confidentiality. Remaining data can be used to re-identify individuals by linking or matching the data to other database, or by looking at unique characteristics found in the database. A formal model based on Modal logic is the theoretical foundation of Cellsecu. As well, a new confidentiality criteria called "non-uniqueness" is defined and implemented. We believe modeling this problem formally can clarify the issue as well as clearly identify the boundary of current technology. Base on our preliminary performance evaluation, the confidentiality check module and the confidentiality enhancing module only slightly degrade system performance. PMID- 12909155 TI - Does the physician order-entry system increase the revenue of a general hospital? AB - OBJECTIVE: The purpose of this study was to examine whether the physician order entry system (POE) could increase the outpatient and inpatient revenue of hospitals. METHOD: We analyzed the inpatient and outpatient revenue data of all general hospitals (212) in South Korea obtained from the Korean National Health Insurance Corporation (KNHIC) during the period from 1996 to 1999 using the mixed model for repeated measure data. RESULTS: Analysis of the 4-years' panel data showed that both outpatient and inpatient revenues increased significantly after POE introduction. The hospital characteristics significantly influencing inpatient revenue were the number of beds, number of physicians and the tertiary status of a hospital; whereas those for outpatient revenue were the number of beds, number of physicians, the private status of a hospital, the tertiary status of a hospital and the urban status of a hospital. CONCLUSION: The revenues from both outpatients and inpatients were found to be increased after the introduction of the POE, while controlling for population size, competition, income, hospital location, hospital size, tertiary status and public status. PMID- 12909156 TI - Personal relationships with an intelligent interactive telephone health behavior advisor system: a multimethod study using surveys and ethnographic interviews. AB - The burgeoning of consumer health informatics and virtual health care can help people improve their health. However, little is known about individuals' reactions to such systems. We conducted an evaluation of the telephone-linked care (TLC) system, a computer-based telecommunications system, that functions as an at home monitor, educator, and counselor for patients with chronic health conditions. Our multimethod assessment of individuals' reactions to using TLC included both quantitative and qualitative methods. Ethnographic in-depth open ended interviews indicated more subtle and surprising reactions to TLC than the overall positive responses from surveys: individuals formed personal relationships with this technology. This relationship formation suggests that TLC designers may have been successful in their attempts to emulate a conversation with a human being. Our study adds to evidence that technology can serve as a projective device for peoples' values and psychological issues. Both designers and users project values and goals onto computer-based technologies and take on different identities through it. Different groups of users, therefore, may see the same technology differently. People also form relationships with technologies, as they did with TLC. These findings, as well as implications for system design and health outcomes, need to be explored in additional studies. PMID- 12909157 TI - Applying methodology to electronic medical record selection. AB - BACKGROUND: Given the potential for electronic medical records (EMRs) to influence every aspect of health care, there has been surprisingly little rigorous research applied to this emerging health technology. An initial phase of the COMPETE (computerization of medical practices for the enhancement of therapeutic efficacy) program, which investigates the impact of EMRs and electronic decision support on efficiency, quality of care and privacy concerns, developed and used a rigorous approach to EMR selection. METHODS: A multidisciplinary team with clinical, technical and research expertise led an eight-stage evaluation process with direct input from potential clinical users at each stage. An iterative sequence of review of EMR specifications and features, live product demonstrations, site visits, and negotiations with vendors led to a progressive narrowing of the field of eligible EMR systems. Final scoring was based on three main themes of clinical usability, data quality and vendor maturity issues. RESULTS: The field of eligible EMR systems was relatively easily narrowed to a few finalists. Determination of the important strengths and weaknesses of these systems' usability, data extraction capabilities and vendor viability, required the full suite of evaluation steps. Preferences varied amongst clinicians and between clinicians and technical specialists, particularly regarding the importance of user interface versus database issues. However, the final scoring process showed consensus amongst clinical and technical experts. CONCLUSION: A rigorous, multidisciplinary process is useful in EMR selection. While prolonged and laborious, it is transparent, enhances buy in and realism of expectations by multiple potential users and IT support staff and may, therefore, improve the ultimate success of any EMR implementation project. PMID- 12909158 TI - An integrated medical image database and retrieval system using a web application server. AB - We developed an Integrated Medical Image Database and Retrieval System (INIS) for easy access by medical staff. The INIS mainly consisted of four parts: specific servers to save medical images from multi-vendor modalities of CT, MRI, CR, ECG and endoscopy; an integrated image database (DB) server to save various kinds of images in a DICOM format; a Web application server to connect clients to the integrated image DB and the Web browser terminals connected to an HIS system. The INIS provided a common screen design to retrieve CT, MRI, CR, endoscopic and ECG images, and radiological reports, which would allow doctors to retrieve radiological images and corresponding reports, or ECG images of a patient simultaneously on a screen. Doctors working in internal medicine on average accessed information 492 times a month. Doctors working in cardiological and gastroenterological accessed information 308 times a month. Using the INIS, medical staff could browse all or parts of a patient's medical images and reports. PMID- 12909159 TI - A practical method of linking data from Medicare claims and a comprehensive electronic medical records system. AB - BACKGROUND: Linking administrative and clinical databases provides opportunities for richer studies to improve healthcare, but linkage may require sophisticated algorithms. Linking US Medicare data with large databases used for everyday clinical practice is seldom described in detail in medical literature. OBJECTIVES: Test a deterministic method of linking data from a local electronic medical records system to Medicare data, and report specific details of the algorithm used as well as lessons learned from the linkage process. SUBJECTS: Medicare beneficiaries with medical encounters in selected Indiana counties in the 5-year period ending in 1999. RESULTS: For 6,388 beneficiaries with Medicare data indicating inpatient encounters in the system, 98% had links to the clinical database. Of 7,231 patients hospitalized and registered in the local clinical system, 86% contained a link to Medicare data, and 69% contained a link even without using Social security number (SSN) as an identifier. Medicare data that conflicted with local hospital records by indicating no local hospitalization occurred in 1.8%. More than 2,000 claims contained hospital identifiers that did not exist in the hospital codebook. CONCLUSIONS: Details of a practical, deterministic method of linking Medicare claims to a large electronic records system have been applied and described. Most records were linked without SSN. A variety of inconsistencies were found and these, along with missing or incomplete data, can influence linking. Integrity of specific variables must be assessed carefully. PMID- 12909160 TI - Context-based ontology building support in clinical domains using formal concept analysis. AB - OBJECTIVE: Ontology in clinical domains is becoming a core research field in the realm of medical informatics. The objective of this study is to explore the potential role of formal concept analysis (FCA) in a context-based ontology building support in a clinical domain (e.g. cardiovascular medicine here). METHODOLOGY: We developed an ontology building support system that integrated an FCA module with a natural language processing (NLP) module. The user interface of the system was developed as a Protege-2000 JAVA tab plug-in. A collection of 368 textual discharge summaries and a standard dictionary of Japanese diagnostic terms (MEDIS ver2.0) were used as the main knowledge sources. A preliminary evaluation was taken to show the usefulness of the system. RESULTS: Stability was shown on the MEDIS-based medical concept extraction with high precision. 73+/-14% (mean+/-S.D.) of the compound medical phrases extracted were sufficiently meaningful to form a medical concept from a clinical perspective. Also, 57.7% of attribute implication pairs (i.e. medical concept pairs) extracted were identified as positive from a clinical perspective. CONCLUSION: Under the framework of our ontology building support system using FCA, the clinical experts could reach a mass of both linguistic information and context-based knowledge that was demonstrated as useful to support their ontology building tasks. PMID- 12909161 TI - Empowerment of patients and communication with health care professionals through an electronic health record. AB - OBJECTIVE: The aim of this project was to design and develop a personal electronic health record (EHR) in order to support patient empowerment and additionally to enhance their communication and information exchange with health professionals through this EHR. METHOD: The functionality of a personal Electronic Healthcare Record (EHR) may vary from a simple web-based interface for interactive data entry and data review up to a much more powerful system additionally supporting electronic data/document communication between clinical information systems of primary care practitioners or hospitals and even reminder based support for the empowered citizen, to actively take care of his health, based on relevant disease management programs. It is one means to support patient empowerment, additionally supported by tools for building a patient community. Since storage and communication of data in an EHR comprises sensible personal health data, each of those functions needs specific security and access management requirements to be considered and implemented. RESULT: Clinical pilot projects are already done or under development. PMID- 12909162 TI - XML as standard for communicating in a document-based electronic patient record: a 3 years experiment. AB - During the past few years, the eXtensible Markup Language (XML) has progressively become a gold standard for accessing, representing and exchanging information, especially in the health care environment. This paper presents an implementation of the use of XML for the electronic patient record (EPR) and discusses more specifically its growing use in two areas of the EPR: first, as a format for the exchange of structured messages, and second, as a comprehensible way of representing patient documents. These statements rely on a 3 years experiment conducted at the Geneva University Hospital as part of its document-centered EPR. PMID- 12909163 TI - Data security and protection in cross-institutional electronic patient records. AB - This paper aims at identifying the specific legal requirements concerning data security and data protection of patient health data that apply to a cross institutional electronic patient record (EPR) and describes possible solutions for meeting these requirements. In Germany, the legal framework for such records provide that disclosure of patient health information to physicians of third party institutions is only allowed in case that it is necessary for the joint treatment of the patient, i.e. in case of a "treatment connection". As a first step, the functionality of a remote-access architecture was proven allowing a one way connection between the EPR systems of two health institutions in Germany, which jointly treat tumor patients. Besides, a signature system model for ensuring the integrity and authenticity of medical documents was developed and implemented in the existing information system architecture of the University Medical Center of Heidelberg. Especially in Germany, the legal framework for cross-institutional EPRs is very complex and has a considerable influence on the development and implementation of cross-institutional EPRs. However, its introduction is thought to be valuable, since a cross-institutional EPR will improve communication within shared care processes, and thus improve the quality of patient care. PMID- 12909164 TI - Developing a virtual patient record using XML and web-based workflow technologies. AB - Virtual patient records (VPR) provide a means for integrated access to patient information that may be scattered around different healthcare settings. Within the boundaries of a health district providing all levels of care, this concept can be implemented in a corporate Intranet environment to support longitudinal patient care activities across the participating healthcare providers. In this context, a VPR implementation enables autonomous healthcare providers to operate in a cooperative working environment and apply continuity of care. Workflow systems bring this collaboration and cooperation into effect by automatically routing the medical information needed by authorized actors in a healthcare process. This paper presents a VPR framework that allows integrating geographically dispersed medical information within a health district and enhancing collaboration and coordination of authorized workgroups by means of a web-based workflow system. An implementation of the proposed framework is also presented. PMID- 12909165 TI - Towards a PropeR combination of patient records and protocols. AB - PROBLEM: The combination of a computer-based patient record and a decision support system (DSS) may give physicians the decisive push they need to accept such systems. In the PropeR-project we determine the requirements for a generic interface between both these systems and evaluate its potential impact on patient care. This article reports results from the first year in one of the domains under study. It also provides background information about the project, including design considerations and experimental approach of forthcoming years. PURPOSE: The objectives of the first year were to determine bottlenecks of the current situation and to determine expected improvements and conditions for implementation of a future situation with computer support. METHODS: These topics were investigated in general by literature review and in the local situation by a requirements analysis. For the analysis we used a combination of observation, interviews, and patient record study. For the literature survey we searched for reviews, meta-analyses and original studies concerning experiences with computer based patient records and DSSs in conventional settings and in integrated care. RESULTS: Main bottleneck was the poor use of shared patient record and paper guidelines. Expected improvements were better protocol adherence and communication. Conditions for implementation of computer support were a proper system interface and adoption by the user. CONCLUSION: Our main conclusion was that the aspect of cooperation needs more decision support than the clinical work itself. PMID- 12909166 TI - PropeR: a multi disciplinary EPR system. AB - This article describes the architecture of an EPR system developed for the PropeR project. This EPR system not only aims at supporting home care of stroke patients, but is also designed in such a way that it can be ported to other medical services without much effort. We will briefly describe the Stroke Service and the related PropeR project. Starting from a list of requirements to construct a generic EPR system we will outline the architecture and describe the standards and methods used. Subsequently we describe the implementation and the problems encountered. In the discussion, we will go into the advantages and disadvantages of the tools and techniques we have used. PMID- 12909167 TI - Healthcare teams over the Internet: programming a certificate-based approach. AB - Healthcare environments are a representative case of collaborative environments since individuals (e.g. doctors) in many cases collaborate in order to provide care to patients in a more proficient way. At the same time modern healthcare institutions are increasingly interested in sharing access of their information resources in the networked environment. Healthcare applications over the Internet offer an attractive communication infrastructure at worldwide level but with a noticeably great factor of risk. Security has, therefore, become a major concern. However, although an adequate level of security can be relied upon digital certificates, if an appropriate security model is used, additional security considerations are needed in order to deal efficiently with the above team-work concerns. The already known Hybrid Access Control (HAC) security model supports and handles efficiently healthcare teams with active security capabilities and is capable to exploit the benefits of certificate technology. In this paper we present the way for encoding the appropriate authoritative information in various types of certificates, as well as the overall operational architecture of the implemented access control system for healthcare collaborative environments over the Internet. A pilot implementation of the proposed methodology in a major Greek hospital has shown the applicability of the proposals and the flexibility of the access control provided. PMID- 12909168 TI - A process for specifying integration for multi-tier applications in healthcare. AB - Integration of the heterogeneous application systems in healthcare is needed to support the clinical and administrative work, patient-centric care, regional interoperability and utilisation of both valuable legacy systems and new technologies. The integration activity is complicated, and must be supported by flexible processes and methods. The integration methods should support the evaluation and specification of different integration approaches, standards and technologies on many interoperability levels. We introduce a process for specifying many integration decisions for a given integration situation. The process is part of an integration method, which is validated in PlugIT project in Finland. PMID- 12909169 TI - Qumquad: a UML-based approach for remodeling of legacy systems in health care. AB - OBJECTIVE: Health care information systems still comprise legacy systems to a certain extent. For reengineering legacy systems a thorough remodeling is inalienable. Current modeling techniques like the Unified Modeling Language (UML) do not offer a systematic and comprehensive process-oriented method for remodeling activities. METHOD: We developed a systematic method for remodeling legacy systems in health care called Qumquad. Qumquad consists of three major steps: (i) modeling the actual state of the application system, (ii) systematic identification of weak points in this model and (iii) development of a target concept for the reimplementation considering the identified weak points. RESULTS: We applied Qumquad for remodeling a documentation and therapy planning system for pediatric oncology (DOSPO). As a result of our remodeling activities we regained an abstract model of the system, an analysis of the current weak points of DOSPO and possible (partly alternative) solutions to overcome the weak points. DISCUSSION AND CONCLUSION: Qumquad proved to be very helpful in the reengineering process of DOSPO since we now have at our disposal a comprehensive model for the reimplementation of DOSPO that current users of the system agree on. Qumquad can easily be applied to other reengineering projects in health care. PMID- 12909170 TI - Discharge and referral data exchange using global standards--the SCIPHOX project in Germany. AB - The goal of the German project "Standardization of Communication between Information Systems in Physician Offices and Hospitals using XML" (aka SCIPHOX) in its first phase is to provide information exchange based on the Extended Markup Language XML between Hospital Information Systems (HIS) and Physician Office Systems (POS). The Clinical Document Architecture (CDA), a standard developed by the Health Level Seven organization (HL7), was chosen to serve as the "backbone" specification. The CDA is an ANSI approved document architecture for exchange of clinical information using XML. In phase I of the SCIPHOX project the proposal specifies the use of the CDA as a generalized international standard in the national context of discharge and referral letters in Germany. The specification defines how to use the CDA header and associated vocabularies by providing a translation and interpretation of the CDA header tags and provides a solution for taking local needs (insurance information etc.) into account. PMID- 12909171 TI - Collaboration--a new IT-service in the next generation of regional health care networks. AB - During the past 10-15 years, Regional Health Care Networks (RHCN) have been established in many regions throughout the world. RHCN build on well-known techniques, methodologies and appropriate standards. Most of the European Countries today have set up IT strategic plans that focus on the establishment of RHCN. The benefits of having access to all relevant information are tremendous and contribute to cost-effective and coherent health services. By the rapid spread and use of Internet, technology has made it possible to interconnect all kinds of applications. In 2000, the most experienced regions in Europe joined PICNIC, a European project to develop the Next Generation Regional Health Care Networks and to support their new ways of providing health and social care. The previous generation of Regional Health Care Networks supported the interconnection of applications by transfer of messages. Messaging is an effective means of integration for isolated high-specialised systems that only need to exchange data. This service will continue to be one of the most important services in the future health care networks. However, tighter coupling may be desirable in some instances to avoid replicating the same functionality in several applications. In other words, certain services can be common and used by a number of applications instead of building that service inside each application. These common services are called middleware services. In PICNIC (http://www.medcom.dk/picnic), a new middleware Collaboration IT service has been identified and developed. This service allows the end users to perform real-time clinical collaboration, with exchange of text, structured data, voice and images across the limits of a single region. A clinical collaboration is associated with the shared clinical context to provide a record of relevant clinical information and facilitates synchronous as well as asynchronous collaboration. This new IT service builds on the increasing popularity of instance messaging and presence systems that facilitate smooth transition between synchronous and asynchronous interaction. The new Collaboration IT service is expected to have a strong impact on the practice of health care in the next generation of Regional Health Care Networks. PMID- 12909172 TI - Case mix use in 25 countries: a migration success but international comparisons failure. AB - Diagnosis related groups (DRGs) were the first health management tool to group patients in clinical meaningful categories with homogeneous resources consumption. They belong to case mix measures that have been introduced in the USA in the seventies to identify clinical differences in groups of patients as well as to document variation in hospitals and physicians practices by comparing similar groups with each others. They are also used to adjust payment rates and to improve quality of care. This innovative concept migrated from the USA to several Western European countries and to Australia first, to Eastern Europe and Asia soon thereafter. The main objective in most countries concerns a hospital financing new methodology, based on similar groups of patients, by applying a global budget. The main result of this international overview of 25 countries is to show the difficulty to make international comparisons, mainly because of the absence of a universal classification system for procedures. Furthermore, diagnostic classification systems to group patients vary also by country. These obstacles to compare hospital and physician practices have profound roots in cultural differences and in political sensitivity for organising health care delivery between countries. The development of an appropriate Case Mix grouper tool including uniform diagnoses and procedures codes, leading to international use becomes a public health priority. PMID- 12909173 TI - The diagnosis related groups enhanced electronic medical record. AB - PROBLEM: The introduction of Diagnosis Related Groups as a basis for hospital payment in Germany announced essential changes in the hospital reimbursement practice. A hospital's economical survival will depend vitally on the accuracy and completeness of the documentation of DRG relevant data like diagnosis and procedure codes. In order to enhance physicians' coding compliance, an easy-to use interface integrating coding tasks seamlessly into clinical routine had to be developed. A generic approach should access coding and clinical guidelines from different information sources. METHODS: Within the Electronic Medical Record (EMR) a user interface ('DRG Control Center') for all DRG relevant clinical and administrative data has been built. A comprehensive DRG-related web site gives online access to DRG grouping software and an electronic coding expert. Both components are linked together using an application supporting bi-directional communication. Other web based services like a guideline search engine can be integrated as well. RESULTS: With the proposed method, the clinician gains quick access to context sensitive clinical guidelines for appropriate treatment of his/her patient and administrative guidelines for the adequate coding of the diagnoses and procedures. This paper describes the design and current implementation and discusses our experiences. PMID- 12909174 TI - Using n-gram method in the decomposition of compound medical diagnoses. AB - OBJECTIVE: Our goal in this study was to find an easy to implement method to detect compound medical diagnosis in Hungarian medical language and decompose them into expressions referring to a single disease. METHODS: A corpus of clinical diagnoses extracted form discharge reports (3,079 expressions, each of them referring to only one disease) was represented in an n-gram tree (a series of n consecutive word). A matching algorithm was implemented in a software, which is able to identify sensible n-grams existing both in test expressions and in the n-gram tree. A test sample of another 92 diagnoses was decomposed by two independent humans and by the software. The decompositions were compared with measure the recall and the precision of the method. RESULTS: There was not full agreement between the decompositions of the humans, (which underlines the relevance of the problem). A consensus was arrived in all disagreed point by a third opinion and open discussion. The resulting decomposition was used as a gold standard and compared with the decomposition produced by the computer. The recall was 82.6% the precision 37.2%. After correction of spelling errors in the test sample the recall increased to 88.6% while the precision slightly decreased to 36.7%. CONCLUSION: The proposed method seems to be useful in decomposition of compound diagnostic expressions and can improve quality of diagnostic coding of clinical cases. Other statistical methods (like vector space methods or neural networks) usually offer a ranked list of candidate codes either for single or compound expressions, and do not warn the user how many codes should be chosen. We propose our method especially in a situation where formal NLP techniques are not available, as it is the case with scarcely spoken languages like Hungarian. PMID- 12909175 TI - Can evaluation studies benefit from triangulation? A case study. AB - BACKGROUND: Information and communication technologies (ICTs) are increasingly being used in health care. Rigorous evaluations of ICT applications during both introduction and routine use are of great importance for decision makers and users. Within evaluation research, two main (and often rather distinct) traditions can be found: the objectivistic and the subjectivistic tradition. METHODS: The theory of triangulation deals with the integration of methods and approaches as to conduct better evaluation studies. In evaluation research, triangulation in general means the multiple employment of various sources of data, observers, methods, and/or theories in investigations of the same phenomenon. We applied triangulation aspects in the analysis of the effects of a computer-based nursing documentation system. RESULTS: We discuss, based on this case study, what benefits can be obtained from applying triangulation in an evaluation study. We show how both the validation of results and the completeness of results can be supported by triangulation. DISCUSSION: The decision whether triangulation may be useful for a given research question, and how it may be correctly applied, requires-like other evaluation methods-intensive training and methodological experience. Medical informatics evaluation research may profit from this well-established theory. PMID- 12909176 TI - Enhancing evidence-based practice--a controlled vocabulary for nursing practice and research. AB - Evidence-based practice requires flexible systems for information retrieval. Search processes in electronic databases must be based on terms, which are familiar to professionals and describe accurately the problem to be solved. The paper describes the process of evolving a controlled nursing vocabulary for indexing and information retrieval purposes. The vocabulary is based on terms of the Medline thesaurus the medical subject headings (MeSH) by National Library of Medicine and the International Classification for Nursing Practice by the International Council of Nurses. The created vocabulary consists of 10 themes central to nursing domain. The content was compiled using Delphi technique. The expert panel had four rounds and at the first stage the vocabulary consists of 558 terms. The validity of the vocabulary has been tested for indexing nursing research and based on the validation the expert panel has made some changes for the hierarchy and content. The vocabulary will be added as an independent theme to the Finnish thesaurus the FinMeSH. PMID- 12909177 TI - Automatic concept extraction from spoken medical reports. AB - OBJECTIVE: The objective of this project is to investigate methods whereby a combination of speech recognition and automated indexing methods substitute for current transcription and indexing practices. METHODS: We based our study on existing speech recognition software programs and on NOMINDEX, a tool that extracts MeSH concepts from medical text in natural language and that is mainly based on a French medical lexicon and on the UMLS. For each document, the process consists of three steps: (1) dictation and digital audio recording, (2) speech recognition, (3) automatic indexing. The evaluation consisted of a comparison between the set of concepts extracted by NOMINDEX after the speech recognition phase and the set of keywords manually extracted from the initial document. The method was evaluated on a set of 28 patient discharge summaries extracted from the MENELAS corpus in French, corresponding to in-patients admitted for coronarography. RESULTS: The overall precision was 73% and the overall recall was 90%. Indexing errors were mainly due to word sense ambiguity and abbreviations. A specific issue was the fact that the standard French translation of MeSH terms lacks diacritics. A preliminary evaluation of speech recognition tools showed that the rate of accurate recognition was higher than 98%. Only 3% of the indexing errors were generated by inadequate speech recognition. DISCUSSION: We discuss several areas to focus on to improve this prototype. However, the very low rate of indexing errors due to speech recognition errors highlights the potential benefits of combining speech recognition techniques and automatic indexing. PMID- 12909178 TI - Electronic patient records: domain message information model perinatology. AB - PURPOSE: The Netherlands is developing a set of national domain information models to support electronic information exchange and electronic patient records (EPR). These domain information models aim to support the development, adoption, implementation and maintenance of the EPR in Dutch healthcare practice. This article describes the modelling for a pilot for mother- and childcare (perinatology). METHODOLOGY: Cases' from perinatology are modelled using the Health Level 7 version 3 Reference Information Model (HL7 RIM) as the methodology and tools. RESULTS: Results include descriptions of care processes, communication and information that are broken down into interaction tables and tables with information. Next several domain information models for perinatology are drawn up. These models allow healthcare professionals to recognise their communication, content and work. Currently, the models facilitate discussion and critique by clinician and informaticians. CONCLUSION: The perinatology domain information models facilitate in building implementations because they contain sufficient details for EPR developers and for developers of messages for information exchange. The first results of the project are useful, despite the fact that HL7 RIM modelling methodology is still not finalized. The approach bridges professional content, technical implementation of messages, and future EPR development. PMID- 12909179 TI - Development of virtual reality stimuli for force platform posturography. AB - People relying much on vision in the control of posture are known to have an elevated risk of falling. Dependence on visual control is an important parameter in the diagnosis of balance disorders. We have previously shown that virtual reality (VR) methods can be used to produce visual stimuli that affect balance, but suitable stimuli need to be found. In this study, the effect of six different VR stimuli on the balance of 22 healthy test subjects was evaluated using force platform posturography. We report in more detail and expand the results published earlier. According to the tests two of the stimuli have a significant destabilizing effect on balance. In addition a significant displacement effect on the subject's center of pressure (COP) was found. Thus it is shown that the design of VR stimuli to cause different effects on the control of balance is possible. PMID- 12909180 TI - Development of diagnostic reference frames for seizures. Part 1: inter participant agreement in the selection of symptoms. AB - OBJECTIVE: Our aim is to develop reliable descriptions of various seizure types, which will be used as a basis for decision support. We use expert opinions in this process. In this contribution we evaluate the inter-participant agreement in the selection of frequently occurring symptoms for the description of seizure types. METHOD: We compared the actual agreement among participants with the agreement that would result from random symptom selection as well as with the maximal agreement attainable. For each seizure type we calculated the reliability coefficients of the responses. RESULTS: For all seizure types we found that the agreement in symptom selection among the participants is significantly higher than expected by chance, but not reaching the maximum agreement attainable. The reliability coefficients varied between 0.56 and 0.74 for the various seizure types. CONCLUSION: Although the participants do not reach the maximum agreement attainable in the selection of symptoms, the majority agreement on characteristic frequently occurring symptoms for the different seizure types does approach the maximum agreement attainable. Therefore, we conclude that expert opinions can be used for building descriptions of seizure types. However, to derive a reliable set of symptoms for the construction of the diagnostic reference frames (DRFs) more participants are needed. PMID- 12909182 TI - Guideline validation in multiple trauma care through business process modeling. AB - Clinical guidelines can improve the quality of care in multiple trauma. In our Department of Trauma Surgery a specific guideline is available paper-based as a set of flowcharts. This format is appropriate for the use by experienced physicians but insufficient for electronic support of learning, workflow and process optimization. A formal and logically consistent version represented with a standardized meta-model is necessary for automatic processing. In our project we transferred the paper-based into an electronic format and analyzed the structure with respect to formal errors. Several errors were detected in seven error categories. The errors were corrected to reach a formally and logically consistent process model. In a second step the clinical content of the guideline was revised interactively using a process-modeling tool. Our study reveals that guideline development should be assisted by process modeling tools, which check the content in comparison to a meta-model. The meta-model itself could support the domain experts in formulating their knowledge systematically. To assure sustainability of guideline development a representation independent of specific applications or specific provider is necessary. Then, clinical guidelines could be used for eLearning, process optimization and workflow management additionally. PMID- 12909181 TI - Development of diagnostic reference frames for seizures. Part 2: are seizure descriptions discriminative? AB - OBJECTIVE: To determine whether the seizure descriptions given by a group of neurologists/epileptologists are discriminative. METHOD: We constructed templates for various seizure types describing how often symptoms were selected by the participants. We defined a matching score to indicate the match between such a template and the symptoms selected by each neurologist/epileptologist individually and computed the scores for each of the sets of selected symptoms with all templates. Correlation coefficients were calculated between the templates. RESULTS: Data were collected from 24 participants. The matching scores and the correlation coefficients both show that participants provide discriminative descriptions of the seizure types. Descriptions of aggregated seizure types, such as primary generalized seizures, are less discriminatory than the descriptions of more specific seizure types. CONCLUSION: We concluded that the participants in our study selected symptoms that result in discriminative descriptions of the seizure types. This indicates that knowledge elicitation by using the opinions of a group of clinical experts is possible. The study also indicates that the design of the study could be ameliorated in several ways. These findings will be taken into account when designing the final study. PMID- 12909183 TI - Which clinical decisions benefit from automation? A task complexity approach. AB - OBJECTIVE: To describe a model for analysing complex medical decision making tasks and for evaluating their suitability for automation. METHOD: Assessment of a decision task's complexity in terms of the number of elementary information processes (EIPs) and the potential for cognitive effort reduction through EIP minimisation using an automated decision aid. RESULTS: The model consists of five steps: (1) selection of the domain and relevant tasks; (2) evaluation of the knowledge complexity for tasks selected; (3) identification of cognitively demanding tasks; (4) assessment of unaided and aided effort requirements for this task accomplishment; and (5) selection of computational tools to achieve this complexity reduction. The model is applied to the task of antibiotic prescribing in critical care and the most complex components of the task identified. Decision aids to support these components can provide a significant reduction of cognitive effort suggesting this is a decision task worth automating. CONCLUSION: We view the role of decision support for complex decision to be one of task complexity reduction, and the model described allows for task automation without lowering decision quality and can assist decision support systems developers. PMID- 12909184 TI - Developing the ontological foundations of a terminological system for end-stage diseases, organ failure, dialysis and transplantation. AB - The Etablissement francais des Greffes (EfG) is a state agency dealing with Public Health issues related to organ, tissue and cell transplantation in France. The evaluation of organ retrieval and transplantation activities, one of its missions, is supported by a national information system (EfG-IS). The EfG-IS is moving towards a new n-tier architecture comprising a terminology server for end stage diseases, organ failure, dialysis and transplantation (EfG-TS). Following a preliminary audit of the existing coding system and in order to facilitate data recording, to improve the quality of information, to assume compatibility with terminological existing standards and to allow semantic interoperability with other local, national or international registries, a specific work has been conducted on the thesauri to integrate within the EfG-TS. In this paper focusing on the server's content rather than the container, we report first the functional and cognitive requirements that resulted from the preliminary audit. We then describe the methodological approach used to build the terminological server on "sound ontological foundations". We performed the semantic analysis of existing medical terms to set up disease description frame-like structures. These diseases description frames consist of a limited set of nosological discriminating slots such as etiology, semiology, pathology, evolution and associated diseases. Each relevant medical term is thus associated to a concept defined and inserted within a hierarchy according to disease description frame resulting from the semantic analysis. Last, because this terminological server is shared by various transplant and dialysis centers to record patient data at different time point, contextualization of terms appeared as one of the functional requirements. We will also point out various contexts for medical terms and how they have been taken into account. PMID- 12909185 TI - Step-by-step mark-up of medical guideline documents. AB - Approaches to formalization of medical guidelines can be divided into model centric and document-centric. While model-centric approaches dominate in the development of clinical decision support applications, document-centric, mark-up based formalization is suitable for application tasks requiring the 'literal' content of the document to be transferred into the formal model. Examples of such tasks are logical verification of the document or compliance analysis of health records. The quality and efficiency of document-centric formalization can be improved using a decomposition of the whole process into several explicit steps. We present a methodology and software tool supporting the step-by-step formalization process. The knowledge elements can be marked up in the source text, refined to a tree structure with increasing level of detail, rearranged into an XML knowledge base, and, finally, exported into the operational representation. User-definable transformation rules enable to automate a large part of the process. The approach is being tested in the domain of cardiology. For parts of the WHO/ISH Guidelines for Hypertension, the process has been carried out through all the stages, to the form of executable application, generated automatically from the XML knowledge base. PMID- 12909186 TI - The effect of alcohol on blood glucose in Type 1 diabetes--metabolic modelling and integration in a decision support system. AB - INTRODUCTION: We have recently shown, in studies with patients with Type 1 (insulin dependent) diabetes, that alcohol intake at 21:00 h significantly reduced blood glucose values after 10-12 h, compared with control studies with no alcohol. HYPOTHESIS: We hypothesised that this was due to the following effects of alcohol: (1) alcohol metabolism increases NADH, leading to a reduction in hepatic gluconeogenesis; (2) increased glycogen phosphorylase activity depletes hepatic glycogen stores; (3) after the alcohol is metabolised, hepatic insulin sensitivity is increased, leading to the restoration of glycogen stores and reduction in blood glucose levels; and (4) consequently, after several hours, glycogen stores and insulin sensitivity return to normal. RESULTS: A model describing these changes (DiasNet-Alcohol) was implemented into the DiasNet model of human glucose metabolism. Our study suggests that the DiasNet-Alcohol model gives a reasonable approximation of these effects of alcohol on blood glucose concentration observed in our study and supports our hypothesis for the mechanism behind these effects in Type 1 diabetes. PMID- 12909187 TI - Collaborative environment for clinical reasoning and distance learning sessions. AB - BACKGROUND: The medical curriculum has changed with the adoption of the student centered learning paradigm. Clinical reasoning learning (CRL) is used in order to develop and improve students' clinical reasoning and problem-solving skills. PURPOSE: We have observed that, in complement to traditional CRL sessions, students commonly consult resources available on the internet. Based on this observation, our objective is to create computer tools to coordinate CRL sessions at distance, integrating these electronic resources at every step of the reasoning process. MATERIAL AND METHODS: In order to create the system, we elaborated an object-oriented model of a computer-supported collaborative learning environment. The proposed system includes a local web-server to store electronic resources and a relational database to store their electronic addresses (urls). JAVA was used as the programming language. RESULTS: We developed a set of cooperative platform-independent tools. This environment includes a communication tool. Multimedia data exchange is possible. Information is shared thanks to an electronic notepad and whiteboard tools. PERSPECTIVES: This learning environment will be integrated in the French Virtual Medical University project, and is intended to be used for undergraduate, internships, residency or continuing medical education. PMID- 12909188 TI - Copability, coping, and learning as focal concepts in the evaluation of computerised diabetes disease management. AB - INTRODUCTION: Within diabetes care, the majority of health decisions are in the hands of the patient. Therefore, the concepts of disease management and self-care represent inescapable challenges for both patient and healthcare professionals, entailing a considerable amount of learning. Thus, a computerised diabetes disease management systems (CDDM) is to be seen not merely as tools for the medical treatment, but also as pedagogical tools to enhance patient competence. HYPOTHESIS: The unfortunate lack of success for most knowledge-based systems might be related to the problem of finding an adequate way of evaluating the systems from their development through the implementation phase to the daily clinical practice. The following presents the initial methodological considerations for evaluating the usefulness of a CDDM system called DiasNet, which is being implemented as a learning tool for patients. The evaluation of usefulness of a CDDM, we claim, entails clinical assessment taking into account the challenges and pitfalls in diabetes disease management. RESULTS: Drawing on activity theory, we suggest the concept of copability as a supplement to 'usability' and 'utility' when determining 'usefulness'. We maintain that it is necessary to ask how well the user copes with the new situation using the system. As ways to measure copability of DiasNet the concepts of coping and learning are discussed, as well as ways this methodology might inform systems development, implementation, and daily clinical practice. PMID- 12909189 TI - N,N,N',N'-tetrakis(2-pyridylmethyl)ethylenediamine inhibits ligand binding to certain G protein-coupled receptors. AB - N,N,N',N'-Tetrakis(2-pyridylmethyl)ethylenediamine (TPEN) is used widely in biological systems to chelate certain heavy metals, particularly Zn2+. Here we show that TPEN inhibits ligand binding to certain G protein-coupled receptors and is an antagonist at muscarinic receptors. In intact human neuroblastoma SH-SY5Y cells, the binding of the muscarinic receptor ligand [N-methyl-3H]scopolamine methyl chloride was inhibited by TPEN (Ki approximately 26 microM), as was muscarinic receptor agonist-induced inositol 1,4,5-trisphosphate formation (Ki approximately 26 microM). This antagonism was not due to metal ion chelation, indicating that it resulted from a direct interaction of TPEN with muscarinic receptors. Examination of the effects of TPEN on other receptors in SH-SY5Y cell membrane preparations showed that the binding of the nonpeptide opioid receptor ligand [15,16-3H]diprenorphine was strongly inhibited, whereas binding of [125I]vasoactive intestinal polypeptide was not. This pattern of selectivity was also seen in AR4-2J rat pancreatoma cell membranes, in which TPEN inhibited ligand binding to muscarinic receptors, but not that to cholecystokinin receptors. In conclusion, these data show that TPEN inhibits ligand binding to certain G protein-coupled receptors and exhibits selectivity towards those receptors whose transmembrane helices form the predominant site for ligand interaction. TPEN may have widespread antagonistic activity towards G protein coupled receptors of this kind. PMID- 12909190 TI - Protein kinase A-dependent coupling of mouse prostacyclin receptors to Gi is cell type dependent. AB - The ability of the prostacyclin (IP) receptor agonist cicaprost to activate Gs-, Gq/11- and Gi-mediated cell signalling pathways has been examined in Chinese hamster ovary (CHO) cells and human embryonic kidney 293 (HEK 293) cells expressing the cloned human (hIP) or mouse (mIP) prostacyclin receptor, and compared with data from NG108-15 and SK-N-SH cells that endogenously express rat/mouse and human IP receptors, respectively. Cicaprost stimulated [3H]cyclic AMP production with EC50 values of 1.5-22 nM, and stimulated [3H]inositol phosphate production (EC50 values 49-457 nM) in all but the SK-N-SH cells. Cicaprost failed to inhibit forskolin-stimulated [3H]cyclic AMP production in any of these cell lines. Therefore, although both human and mouse IP receptors couple to Gs and Gq/11-mediated signalling pathways in a cell type-dependent manner, we could find no evidence for IP receptor coupling to Gi. PMID- 12909191 TI - The opposing effects of endothelin-1 and C-type natriuretic peptide on apoptosis of neonatal rat cardiac myocytes. AB - C-type natriuretic peptide (CNP) and endothelin-1 are paracrine peptides with opposing effects on cardiac myocyte contraction and intracellular cGMP production. Elevated levels of both endothelin-1 and CNP are found in patients with congestive heart failure. These factors may be related to positive and negative regulation of cell apoptosis in the failing heart. To evaluate the effect of CNP and endothelin-1 on apoptosis of cardiac myocytes and the possible mechanisms involved, primary cardiac myocytes were prepared from neonatal Sabra rats. Cardiomyocyte apoptosis was evaluated by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) and Annexin V in situ staining. The TUNEL method was used to measure the apoptotic index. CNP and the cGMP derivative, 8-br-cGMP, induced apoptosis of cardiac myocytes. CNP-induced apoptosis could be blocked by HS 142-1 (a mixture of 20-30 kinds of linear beta 1, 6-glucan esterified by capronic acid, an antagonist of type A and B natriuretic peptide receptors), and KT 5823 (C29H25N3O5), the inhibitor of cGMP dependent protein kinase). Alpha-difluoromethylornithine (DFMO), the irreversible inhibitor of ornithine decarboxylase, also induced apoptosis to a similar extent. CNP and 8-br-cGMP caused a marked reduction of intracellular ornithine decarboxylase expression, as determined by Western blot analysis and immunohistochemical assay. Preincubation with endothelin-1 attenuated CNP- and 8 br-cGMP-induced cardiomyocyte apoptosis. Endothelin-1 also antagonized the CNP- and 8-br-cGMP-induced reduction of intracellular ornithine decarboxylase expression. These results suggest that CNP has a proapoptotic effect on neonatal rat cardiac myocytes. The effect is mediated via natriuretic peptide receptors and is due to an elevation of intracellular cGMP, which reduces the expression of intracellular ornithine decarboxylase and probably the production of polyamines. Endothelin-1 protects cardiac myocytes against CNP-induced apoptosis by influencing the cGMP-dependent pathway, and this effect is probably mediated through both a reduction of cGMP and antagonism of the CNP-induced reduction of intracellular ornithine decarboxylase expression. PMID- 12909192 TI - Nonpeptide gastrin releasing peptide receptor antagonists inhibit the proliferation of lung cancer cells. AB - The ability of nonpeptide antagonists to interact with gastrin releasing peptide receptors on lung cancer cells was investigated. PD176252 (3-(1H-Indol-3-yl)-N-[1 (5-methoxy-pyridin-2-yl)-cyclohexylmethyl]-2-methyl-2-[3-(4-nitro-phenyl)-ureido] propionamide) and PD168368 (3-(1H-Indol-3-yl)-2-methyl-2-[3(4-nitro-phenyl) ureido]-N-(1-pyridin-2-yl-cyclohexylmethyl)-propionamide) inhibited specific 125I gastrin releasing peptide binding to NCI-H1299 cells with IC50 values of 20 and 1500 nM, respectively. Similar binding results were obtained using NCI-H157, H345 and N592 human lung cancer cells. PD176252 inhibited the ability of 1 nM bombesin to cause elevation of cytosolic calcium in Fura-2 loaded NCI-H345 or H1299 cells, whereas it had no effect on basal cytosolic calcium. PD176252 antagonized the ability of 10 nM bombesin to cause elevation of c-fos mRNA in NCI-H1299 cells. Also, PD176252 inhibited the ability of 100 nM bombesin to cause tyrosine phosphorylation of focal adhesion kinase in NCI-H1299 cells. Using a [3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide] assay, PD176252 was more potent than PD168368 at inhibiting NCI-H1299 proliferation. Also, 1 microM PD176252 significantly inhibited lung cancer colony number in vitro. PD176252 in a dose-dependent manner inhibited NCI-H1299 xenograft growth in nude mice in vivo. These results indicate that PD176252 is a gastrin releasing peptide receptor antagonist, which inhibits the proliferation of lung cancer cells. PMID- 12909193 TI - Ligand internalization by cloned neuropeptide Y Y5 receptors excludes Y2 and Y4 receptor-selective peptides. AB - In human embryonic kidney-293 (HEK-293) cells, the cloned human neuropeptide Y Y5 receptor saturably internalized agonists, with the rank order of neuropeptide Y (19-23)-[Gly1,Ser3,Gln4,Thr6,Ala31,Aib32,Gln34]human pancreatic polypeptide (neuropeptide Y-Aib-pancreatic polypeptide)>human neuropeptide Y>porcine peptide YY>[Pro34]human peptide YY>[Leu31,Pro34]human peptide YY>>human peptide YY-(3 36). Human pancreatic polypeptide competed [125I]neuropeptide Y binding and internalization in neuropeptide Y Y5 receptor-expressing cells, but itself showed no internalization. The internalization was strongly dependent on temperature. The surface binding, and especially the internalization, of human neuropeptide Y were highly sensitive to the clathrin network inhibitor phenylarsine oxide, and to the cholesterol-complexing antibiotic filipin III. The internalized ligands were present in particles corresponding to secondary endosomes in Percoll gradients, but especially in particles banding with the acid hexosaminidase lysosomal marker. At any temperature tested, internalization of the neuropeptide Y Y5 receptor driven by human neuropeptide Y in HEK-293 cells was much slower than the internalization of the neuropeptide Y Y1 receptor reported in the same cells, or in Chinese hamster ovary (CHO) cells. The neuropeptide Y Y5 receptor subtype could be the metabotropic receptor responding to protracted challenges by neuropeptide Y-like peptides, and its density could be little sensitive to concentration of extracellular agonists. PMID- 12909194 TI - Functional expression of adenosine A2A and A3 receptors in the mouse dendritic cell line XS-106. AB - There is increasing evidence to suggest that adenosine receptors can modulate the function of cells involved in the immune system. For example, human dendritic cells derived from blood monocytes have recently been described to express functional adenosine A1, A2A and A3 receptors. Therefore, in the present study, we have investigated whether the recently established murine dendritic cell line XS-106 expresses functional adenosine receptors. The selective adenosine A3 receptor agonist 1-[2-chloro-6[[(3-iodophenyl)methyl]amino]-9H-purin-9-yl]-1 deoxy-N-methyl-beta-D-ribofuranuronamide (2-Cl-IB-MECA) inhibited forskolin mediated [3H]cyclic AMP accumulation and stimulated concentration-dependent increases in p42/p44 mitogen-activated protein kinase (MAPK) phosphorylation. The selective adenosine A2A receptor agonist 4-[2-[[-6-amino-9-(N-ethyl-beta-D ribofuranuronamidosyl)-9H-purin-2-yl]amino]ethyl]benzene-propanoic acid (CGS 21680) stimulated a robust increase in [3H]cyclic AMP accumulation and p42/p44 MAPK phosphorylation. In contrast, the selective adenosine A1 receptor agonist CPA (N6-cyclopentyladenosine) did not inhibit forskolin-mediated [3H]cyclic AMP accumulation or stimulate increases in p42/p44 MAPK phosphorylation. These observations suggest that XS-106 cells express functional adenosine A2A and A3 receptors. The non-selective adenosine receptor agonist 5'-N ethylcarboxamidoadenosine (NECA) inhibited lipopolysaccharide-induced tumour necrosis factor-alpha (TNF-alpha) release from XS-106 cells in a concentration dependent fashion. Furthermore, treatment with Cl-IB-MECA (1 microM) or CGS 21680 (1 microM) alone produced a partial inhibition of lipopolysaccharide-induced TNF alpha release (when compared to NECA), whereas a combination of both agonists resulted in the inhibition of TNF-alpha release comparable to that observed with NECA alone. Treatment of cells with the adenosine A2A receptor selective antagonists 4-(2-[7-amino-2-(2-furyl)[1,2,4]triazolo[2,3-a][1,3,5]triazin 5ylamino]ethyl)phenol (ZM 241385; 100 nM) and 5-amino-2-(2-furyl)-7-phenylethyl pyrazolo[4,3-e]-1,2,4-triazolo[1,5c]pyrimidine (SCH 58261; 100 nM) and the adenosine A3 receptor selective antagonist N-[9-chloro-2-(2-furanyl)[1,2,4] triazolo[1,5-c]quinazolin-5-benzeneacetamide (MRS 1220; 100 nM) partially blocked the inhibitory effects of NECA on lipopolysaccharide-induced TNF-alpha release. Combined addition of MRS 1220 and SCH 58261 completely blocked the inhibitory effects of NECA on lipopolysaccharide-induced TNF-alpha release. In conclusion, we have shown that the mouse dendritic cell line XS-106 expresses functional adenosine A2A and A3 receptors, which are capable of modulating TNF-alpha release. PMID- 12909196 TI - Behavioral sensitization and alteration in monoamine metabolism in mice after single versus repeated methamphetamine administration. AB - To address the functional alterations of monoaminergic neuronal systems in mice after single and repeated administration of methamphetamine, we examined the tissue contents of monoamines and their metabolites in addition to locomotor activity estimated by horizontal locomotion and rearing measurements. In male ICR mice, the repeated treatment regimen (intraperitoneal administration of 1.0 mg/kg methamphetamine once per day for five consecutive days) induced hyperlocomotion with a plateau level on test day 4. The initial behavioral response (within 5 min after injection) to the drug appeared to include context-dependent sensitization. Mice after the initial repeated treatment regimen showed behavioral sensitization to the same dose of methamphetamine 5 days after the final injection (test day 11). On test day 11, the first 150 min, but not the nocturnal behavior (during the dark hours), were significantly enhanced after the drug challenge. A marked reduction of the content of L-dihydroxyphenylalanine and the ratio of 3,4 dihydroxyphenylacetic acid to dopamine was observed in the striatum+accumbens of mice after single and repeated administration of methamphetamine. As for serotonin metabolism, the ratio of 5-hydroxyindolacetic acid to serotonin significantly increased in mice after single administration of methamphetamine, although it decreased in mice after repeated administration of methamphetamine. Norepinephrine metabolism (the ratio of 3-methoxy-4-hydroxyphenylglycol to norepinephrine) was not affected in the striatum+accumbens or thalamus+hypothalamus of the mice after repeated or single methamphetamine treatment. These results suggest that dopaminergic and serotonergic neuronal activities were altered during the development of behavioral sensitization. The ratio of 3-methoxytyramine to dopamine was not affected, suggesting that the methamphetamine treatment selectively inhibited the monoamine oxidase pathway for dopamine inactivation. PMID- 12909195 TI - In vivo neuroprotective effects of ACEA 1021 confirmed by magnetic resonance imaging in ischemic stroke. AB - The neuroprotective activity of ACEA 1021 (5-nitro-6,7-dichloro-1,4-dihydro-2,3 quinoxalinedione; licostinel), a selective antagonist at the strychnine insensitive glycine site associated with the NMDA receptor complex, has been investigated in various models of focal cerebral ischemia. In isoflurane anaesthesised Wistar rats with permanent ipsilateral carotid artery ligation and transient middle cerebral artery occlusion (duration of occlusion, 2 h) followed by reperfusion (24 h), intravenous administration of ACEA 1021 (bolus: 10 mg/kg, 15 min after the onset of middle cerebral artery occlusion; infusion: 7 mg/kg/h for 6 h beginning 30 min after occlusion of the artery) produced a 32% reduction in infarct volume. Similarly, in Sprague-Dawley rats with transient middle cerebral artery occlusion (2 h) followed by 24 h of reperfusion, identical treatment with ACEA 1021 decreased infarct size by 39%. Magnetic resonance imaging (MRI) confirmed these effects in the transient model, in that infarct volume observed using apparent diffusion coefficient (ADC) maps was significantly smaller after 24 h in the ACEA 1021-treated rats compared with Tris-treated controls. Furthermore, the increase in perfusion signal intensity after reperfusion was more pronounced in the ACEA 1021-treated rats than in controls. In Fisher 344 rats with permanent occlusion of the middle cerebral artery, ACEA 1021 induced a dose-related decrease in infarct volume, which was associated with an improvement in neurological outcome as measured by the rope suspension procedure. Administration of the same dose regimen, as above, in Fisher rats with permanent middle cerebral artery occlusion reduced infarct volume by 68%. This dose was as effective when administration was delayed for 2 h. In mice with permanent middle cerebral artery occlusion, ACEA 1021 (5 mg/kg, i.v., 5 min after occlusion; 30 mg/kg, s.c., 1 and 4 h post-middle cerebral artery occlusion) decreased infarct size by 42%. The consistent anti-ischemic effects of ACEA 1021 make it a valuable compound for exploratory stroke research. PMID- 12909197 TI - Subsensitivity of P2X but not vanilloid 1 receptors in dorsal root ganglia of rats caused by cyclophosphamide cystitis. AB - The application of cyclophosphamide to rats was used to induce interstitial cystitis. Behavioural studies indicated a strong pain reaction that developed within 2 h and levelled off thereafter causing a constant pain during the following 18 h. Neurons prepared from L6/S1 dorsal root ganglia innervating the urinary bladder responded to the application of capsaicin or alpha,beta-methylene ATP (alpha,beta-meATP) with an increase of intracellular Ca2+ ([Ca2+]i). The [Ca2+]i responses to capsaicin were identical in the dorsal root ganglion cells of cyclophosphamide- and saline-treated rats, whereas alpha,beta-meATP induced less increase in [Ca2+]i in the cyclophosphamide-treated animals than in their saline-treated counterparts. Hence, alpha,beta-meATP-sensitive P2X3 and/or P2X2/3 receptors of L6/S1 dorsal root ganglion neurons were functionally downregulated during subacute pain caused by experimental cystitis. In contrast, capsaicin sensitive vanilloid 1 receptors did not react to the same procedure. Thoracal dorsal root ganglia, not innervating the urinary bladder, were also unaltered in their responsiveness to alpha,beta-meATP by cyclophosphamide treatment. PMID- 12909198 TI - Behavioral evidence for mu-opioid and 5-HT2A receptor interactions. AB - Electrophysiological studies have demonstrated a physiological interaction between 5-HT2A and mu-opioid receptors in the medial prefrontal cortex. Furthermore, behavioral studies have found that phenethylamine hallucinogens induce head shakes when directly administered into the medial prefrontal cortex. The receptor(s) by which morphine suppresses head shakes induced by serotonin agonists have not been characterized. We administered mu-opioid receptor agonists and antagonists to adult male Sprague-Dawley rats prior to treatment with the phenethylamine hallucinogen 1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane (DOI), which is known to induce head shakes via 5-HT2A receptors. The suppressant action of the moderately selective mu-opioid receptor agonist, buprenorphine (ID50 approximately 0.005 mg/kg, i.p.; a mu-opioid receptor partial agonist and kappa opioid receptor antagonist) was blocked by naloxone and pretreatment with the irreversible mu-opioid receptor antagonist clocinnamox. Another mu-opioid receptor agonist fentanyl also suppressed DOI-induced head shakes. In contrast, a delta-opioid receptor agonist was without effect on DOI-induced head shakes. Thus, activation of mu-opioid receptors can suppress head shakes induced by hallucinogenic drugs. PMID- 12909199 TI - Behavioral and biochemical investigations of bupropion metabolites. AB - The stimulus effects of bupropion metabolites were examined in a drug discrimination procedure using (-)nicotine- and (+)amphetamine-trained rats. (+)- and (-)threohydrobupropion partially substituted in each group. R,R hydroxybupropion produced vehicle-appropriate responding in (-)nicotine animals but, when given in combination with the training dose of (-)nicotine, resulted in an attenuated effect. S,S-Hydroxybupropion partially (66%) substituted for ( )nicotine. In (+)amphetamine-trained animals, S,S-hydroxybupropion (ED50=4.4 mg/kg) generalized completely and was similar in potency to bupropion (ED50=5.4 mg/kg). Bupropion and its metabolites lacked affinity for nicotinic acetylcholinergic receptors, but all antagonized (-)nicotine-induced 86Rb+ efflux in cells expressing alpha3beta4 nicotinic cholinergic receptors. S,S Hydroxybupropion possessed affinity at the dopamine transporter comparable to bupropion, and was also found to bind at the norepinephrine transporter. Although it is unlikely that any metabolite isomer is chiefly responsible for the stimulus actions of bupropion, some probably play a role in the complex actions of this agent. PMID- 12909200 TI - Involvement of the melanocortin MC4 receptor in stress-related behavior in rodents. AB - The melanocortin subtype 4 (MC4) receptor has been postulated to be involved in stress and stress-related behavior. We made use of melanocortin MC4 receptor agonists and antagonist to investigate the relationship between the melanocortin MC4 receptor and stress related disorders. The nonspecific melanocortin receptor agonist alpha-melanocyte stimulating hormone (alpha-MSH) and the melanocortin MC4 receptor agonist, Ac-[Nle4,Asp5,D-Phe7,Lys10]alpha-MSH-(4-10)-NH2 (MT II) dose dependently and significantly reduced the number of licking periods in the rat Vogel conflict test, suggesting that stimulation of the melanocortin MC4 receptor causes anxiogenic-like activity in rats. We synthesized a peptidemimetic melanocortin MC4 receptor selective antagonist, Ac-D-2Nal-Arg-2Nal-NH2 (MCL0020), which has high affinity for the melanocortin MC4 receptor with IC50 values of 11.63 +/- 1.48 nM, in contrast, the affinities for melanocortin MC1 and MC3 receptors were negligible. In addition, MCL0020 significantly attenuated the cAMP formation induced by alpha-MSH in COS-1 cells expressing the melanocortin MC4 receptor without affecting basal cAMP contents. Thus, we considered MCL0020 to be a selective melanocrotin MC4 receptor antagonist among melanocortin receptors. Restraint stress significantly reduced food intake in rats, and i.c.v. administration of MCL0020 dose-dependently and significantly attenuated restraint stress-induced anorexia without affecting food intake. Swim stress induced reduction in the time spent in the light area in the mouse light/dark exploration test, and MCL0020 significantly prevented it. Taken together our findings suggest that the melanocortin MC4 receptor might be related to stress-induced changes in behavior, and blockade of the melanocortin MC4 receptor may prevent stress induced disorders such as anxiety. PMID- 12909201 TI - Ca2+-activated K+ channels in the endothelial cell layer involved in modulation of neurogenic contractions in rat penile arteries. AB - The present study was designed to investigate the functional K+ channels involved in contractions induced by electrical field stimulation in isolated rat penile arteries. Blockers of Ca2+-activated K+ channels (KCa), tetraethylammonium, and of large-conductance KCa channels, charybdotoxin and iberiotoxin, as well as a blocker of voltage-dependent K+ channels (KV), 4-aminopyridine, increased resting tension in penile small arteries. In the presence of propranolol and NG-nitro-L arginine (L-NOARG), electrical field stimulation evoked prazosin-sensitive contractions. In endothelium-intact preparations, these latter contractions were enhanced in the presence of tetraethylammonium and charybdotoxin. However, these blockers did not enhance contractions evoked by exogenously added noradrenaline. Endothelial cell removal increased the neurogenic contractions but tetraethylammonium had no further potentiating effect in these preparations. In the presence of an inhibitor of cyclooxygenase, indomethacin, and inhibitor of nitric oxide (NO) synthase, L-NOARG, acetylcholine evoked relaxations, which were abolished in the presence of either tetraethylammonium or charybdotoxin. In phenylephrine-contracted arteries treated with guanethidine and atropine, electrical field stimulation evoked relaxations, which were partially inhibited by L-NOARG and tetraethylammonium, without any additive effect of these drugs. These observations suggest that both large-conductance KCa channels and KV channels sensitive to iberiotoxin/tetraethylammonium and 4-aminopyridine, respectively, are directly involved in the modulation of myogenic tone of rat penile arteries. Furthermore, activation of endothelial intermediate-conductance KCa channels sensitive to tetraethylammonium and charybdotoxin leads to release of a non-NO nonprostanoid factor, which inhibits release of the neurotransmitter, noradrenaline, but these channels do not appear to be involved in inhibition of contraction evoked by exogenously applied noradrenaline in rat penile arteries. PMID- 12909202 TI - SL65.0472 blocks 5-hydroxytryptamine-induced vasoconstriction in a dog hindlimb ischemia model. AB - We have studied the ability of SL65.0472 (7-fluoro-2-oxo-4-[2-[4-(thieno[3,2 c]pyridin-4-yl)piperazin-1-yl]ethyl]-1,2-dihydroquinoline-1-acetamide), a 5 hydroxytryptamine (5-HT) 5-HT1B/5-HT2A receptor antagonist, to antagonise the vasoconstrictor effects of 5-HT and sumatriptan in a canine model of hindlimb ischemia. Dogs underwent right external iliac artery ligation and right superficial femoral artery excision, resulting in decreased perfusion (-31%, P<0.05) in the right hindlimb. Following pretreatment with L-NAME, phentolamine and propranolol, intra-aortic injection of 5-HT markedly reduced blood flow to the right ischemic hindlimb (-50 +/- 2%, P<0,05). 5-HT induced vasoconstriction was significantly inhibited (-66%, P<0.05) by SL65.0472 (300 microg/kg i.v.), but unaffected by ketanserin (300 microg/kg i.v.), a 5-HT2A receptor antagonist. SL65.0472 also blocked sumatriptan-induced vasoconstriction in ischemic and normally perfused hindlimbs. Thus, SL65.0472 is an effective antagonist of 5-HT receptor mediated hindlimb vasoconstriction. PMID- 12909203 TI - Herbimycin A attenuates apoptosis during heat stress in rats. AB - Expression of heat shock proteins (HSPs) as a heat stress response is associated with acquisition of thermotolerance. Herbimycin A is a tyrosine kinase inhibitor that has been shown to induce HSPs. The present study aims to investigate the effects of herbimycin A on thermotolerance in rats subjected to heat stress exposure. Herbimycin A induced hsp70 to peak levels 12 h post-injection in rats without heat stress. No change in hsp70 levels was observed in the vehicle- and saline-treated rats. In rats exposed to heat stress at 45 degrees C for 25 min, 12 h post-treatment, lower peak temperatures were attained in herbimycin A treated group as compared to the vehicle- and saline-treated groups. Terminal transferase-mediated d-UTP nick end labeling (TUNEL) showed that a significant decrease in apoptosis of hepatocytes in herbimycin A-treated rats as compared to the vehicle- and saline-treated rats. Caspase-3 activation was also lower in herbimycin A-treated rats, compared to the vehicle- and saline-treated rats. The present study has demonstrated that herbimycin A is effective for development of thermotolerance and therefore protects rats from heat stress. PMID- 12909204 TI - Role of endothelin ETA receptors in sepsis-induced mortality, vascular leakage, and tissue injury in rats. AB - The role of endothelin ETA receptors in sepsis-induced mortality and edema formation was evaluated with a selective antagonist ABT-627 [2-(4-methoxyphenyl) 4-(1,3-benzodioxol-5-yl)-1-(N,N-di(n-butyl)amino carbonylmethyl)-pyrrolidine-3 carboxylic acid]. Sprague-Dawley rats received saline (control group), Escherichia coli endotoxin (10 mg/kg, sepsis group) or infusion of ABT-627 prior and immediately after saline and endotoxin injection. Mortality, edema formation (wet/dry ratios), and multiple tissue injury (indicated by serum concentrations of creatinine, urea, bilirubin, creatine kinase, lactate dehydrogenase, and aspartate aminotransferase) were monitored within 5 h. Endotoxin injection elicited 64% mortality, significantly augmented edema formation in liver, heart, lung, and kidney, and raised serum levels of tissue injury markers. Pretreatment with ABT-627 completely reversed endotoxin-induced mortality, significantly attenuated wet/dry ratios of the heart, liver, and kidney, but not lungs, and reduced serum levels of creatine kinase, creatinine, aspartate aminotransferase, and lactate dehydrogenase, but not that of urea and bilirubin. These results suggest that endothelin ETA receptors play a significant role in promoting mortality, edema formation (except in the lungs), and tissue injury in animals with severe sepsis. PMID- 12909205 TI - History, protocols, and the team. PMID- 12909206 TI - Is team management appropriate for a patient receiving coronary brachytherapy? PMID- 12909207 TI - Is the patient on this team? PMID- 12909208 TI - Migration and myelination. PMID- 12909209 TI - Evaluation of the Houston biochemical relapse definition in men treated with prolonged neoadjuvant and adjuvant androgen ablation and assessment of follow-up lead-time bias. AB - PURPOSE: To validate the Houston prostate-specific antigen relapse definition in a mature cohort of men treated with external beam radiotherapy (EBRT) and adjuvant androgen ablation (AA) and men treated with EBRT monotherapy, and to compare these results with the American Society for Therapeutic Radiology and Oncology (ASTRO) and Vancouver prostate-specific antigen relapse (biochemical no evidence of disease) definitions. METHODS AND MATERIALS: A prospective database of 1490 men treated with EBRT, with or without AA, was examined. The impact on hazard proportions, as well as the predictive ability, of the Houston, ASTRO, and Vancouver definitions was tested. RESULTS: For all patients, the Houston definition was more accurate (79.5%) than the ASTRO (76.7%) or Vancouver (77.2%) definitions in predicting subsequent clinical relapse. The Houston definition was superior to the ASTRO definition in those treated both with and without AA and equivalent to the Vancouver definition in those receiving AA. The Houston definition demonstrated proportional hazards when categorized for the use of AA, unlike the ASTRO and Vancouver definitions. The effect of inadequate follow-up on the projected relapse rates was negligible with the Houston definition. CONCLUSION: The Houston relapse definition is favored after EBRT monotherapy or combined EBRT and AA. Use of the Cox proportional hazard multivariate analysis is appropriate with the Houston definition, but not with the ASTRO or Vancouver definitions if AA and non-AA patients are combined. PMID- 12909210 TI - Biochemical failure as a determinant of distant metastasis and death in prostate cancer treated with radiotherapy. AB - PURPOSE: Biochemical failure (BF), defined by a rising prostate-specific antigen (PSA) profile, is an early surrogate of treatment failure. However, little evidence is available to show that BF is associated with death for patents with prostate cancer treated with radiotherapy. We examined the relationship between BF and death from prostate cancer. METHODS AND MATERIALS: A total of 942 patients were treated between 1987 and 1998 with external beam radiotherapy who had sufficient PSA determinations in follow-up for the analyses described. The median radiation dose was 72 Gy, median PSA was 9.9 ng/mL, and median follow-up was 73 months. The American Society for Therapeutic Radiology and Oncology consensus definition was used to define BF. Kaplan-Meier calculations were from the start of radiotherapy. Cox proportional hazards regression multivariate analyses were used to investigate the association of BF (time-dependent variable) and other factors to distant metastasis (DM), cause-specific death (CSD), and overall death (OD). The year of treatment was included in some of the multivariate analyses to correct for potential unknown factors that may have occurred during the years of the study, such as stage migration. RESULTS: BF was observed in 316 patients (34%), and 66 (7%) experienced DM, 32 (3%) died of prostate cancer, and 230 (24%) died overall during the study period. The Kaplan-Meier 5-year rate estimates from the start of treatment for BF, DM, CSD, and OD were 38%, 6%, 3%, and 13%, respectively. All patients with DM had BF. In multivariate analyses, BF was associated with DM and CSD, but not OD. The inclusion of the year of treatment did not alter these relationships. CONCLUSION: BF, as a time-dependent covariate, was the strongest determinant of DM and was also very significantly related to CSD. The inclusion of the year of treatment had little effect on these associations. Longer follow-up is needed to determine conclusively the relationship of BF to OD. PMID- 12909211 TI - Effect of sequencing of androgen deprivation and radiotherapy on prostate cancer growth. AB - PURPOSE: Androgen deprivation (AD) is frequently combined with radiotherapy (RT); however, the optimal sequence in vivo is currently unknown. Previous published work from our laboratory demonstrated that AD with RT was consistent with at least an additive, and possibly supra-additive, effect with the combined approach. We, therefore, performed additional experiments to elucidate the optimal sequence. METHODS AND MATERIALS: R3327-G Dunning rat prostate tumor cells were grown s.c. in the flanks of Copenhagen rats. Treatment was initiated when the tumor reached approximately 1 cm(3). Temporary AD was performed by a transscrotal orchiectomy followed 14 days later with androgen restoration using s.c. testosterone implants. RT was delivered using (60)Co to 7 Gy. Seven groups, including the controls, were analyzed: Group 1, sham control (Day 0: AD + testosterone); 2, AD control (Day 0: AD, Day 14: testosterone); 3, RT alone on Day 7 (Day 0: AD + testosterone, Day 7 RT); 4, RT alone on Day 3 (Day 0: AD + testosterone, Day 3: RT); 5, RT during AD (Day 0: AD, Day 7: RT, Day 14: testosterone); 6, RT before AD (Day 0: RT, Day 3: AD, Day 17: testosterone); and Group 7, RT after AD (Day 0: AD, Day 14: testosterone, Day 17: RT). The doubling times for tumor growth were calculated for the seven groups from the end of treatment plus 1 day. Differences in doubling time were assessed using analysis of variance, with pair-wise comparisons accomplished using post-hoc Bonferroni tests. RESULTS: An analysis of the differences in the tumor volume doubling time as measured from the end of treatment suggests that Groups 1 and 7 were statistically different from the other groups (p = 0.02). As expected, the sham control group had the shortest doubling time at 5.4 days and Group 7 (14 days of AD administered before RT) had the longest doubling time at 32.6 days. The findings were similar even after excluding an outlying doubling time of 85 days from Group 7 (p < 0.0001). To assess the effect of sequencing further, only Groups 5 through 7 (excluding the outlier) were compared in an analysis of variance with post-hoc Bonferroni tests. Group 7 (RT after AD) demonstrated a significantly longer doubling time than Groups 5 and 6 (p = 0.0024). CONCLUSION: The results suggest that neoadjuvant AD may result in prolonged suppression of tumor growth, even after testosterone replacement. PMID- 12909213 TI - Definitive radiotherapy of prostate cancer: the possible role of staging lymphadenectomy. AB - PURPOSE: To evaluate the postradiotherapy 5-year cancer-specific (CSS), clinical progression-free (cPFS), and overall (OS) survival rates in patients with pN0 M0 prostate cancer (PC). METHODS: Between 1989 and 1996, 203 consecutive pN0 M0 PC patients (T1-2, 66; T3-4, 137) received conformal prostatic four-field radiotherapy (median target dose 66 Gy). Any hormone manipulation was delayed until clinical progression (growth of the primary tumor or development of distant metastases). RESULTS: After a median observation time of 87 months (range 11 156), 99 patients had relapsed clinically and 70 patients were dead, 37 of them as a result of prostate cancer. Five-year CSS, cPFS, and OS rates were, respectively, 90% (95% CI 86-94%), 64% (95% CI 57-71%), and 82% (95% CI 77-87%), with no difference of OS compared with age-matched males from the general population. Gleason score (< or =7A vs. > or =7B) and the T category predicted cPFS, whereas CSS was associated with Gleason score only. Preradiotherapy PSA failed to predict survival. Patients with T1-2 Gleason score < or =7A had a 97% 5 year CCS, as compared with 89% for all other patients. A median of eight lymph nodes (range 0-29) were described in the specimens from pelvic lymphadenectomy (LA). CONCLUSION: Despite still preliminary observations, our 5-year results challenge the use of combined hormone radiotherapy in patients who are proven to be pN0 by preradiotherapy LA; in particular, in patients with T1-2/Gleason score < or =7A, whereas the survival in all other patients with pN0 M0 prostate cancer may be improved by adjuvant androgen deprivation. PMID- 12909212 TI - A comparison of CT scan to transrectal ultrasound-measured prostate volume in untreated prostate cancer. AB - PURPOSE: To compare CT and transrectal ultrasound (TRUS)-measured prostate volumes in patients with untreated prostate cancer. METHODS AND MATERIALS: Between 1995 and 1999, 48 consecutive patients at the Portland Veterans Affairs Medical Center were treated with external beam radiotherapy. In 36 of these patients, TRUS and CT measurements of the prostate volume were obtained before treatment and <6 months apart. The TRUS volume was calculated using the prolate ellipsoid formula. The CT volume was calculated from the contours of the prostate drawn by one physician, who was unaware of the TRUS volume calculation, on axial CT images. RESULTS: The TRUS and CT prostate volume measurements correlated strongly (Pearson's correlation coefficient = 0.925, 95% confidence interval 0.856-0.961, p < 0.0001). The CT volume was consistently larger than the TRUS volume by a factor of approximately 1.5. In men with a TRUS prostate volume less than the median (<28 cm(3)), the CT/TRUS volume ratio was 1.7, and it was 1.4 for men whose volume was greater than the median. The CT volumes were correlated similarly with the TRUS volumes regardless of the CT slice interval. CONCLUSION: A strong correlation was found between CT scan and TRUS measurement of the prostate volume; however, CT consistently overestimated the prostate volume by approximately 50% compared with TRUS. PMID- 12909214 TI - Late rectal function after prostate brachytherapy. AB - PURPOSE: Using a patient-administered quality of life instrument, to evaluate the effect of permanent prostate brachytherapy on late rectal function. METHODS AND MATERIALS: One hundred eighty-nine prostate brachytherapy patients were mailed the Rectal Function Assessment Score (R-FAS), with a prestamped return envelope. The R-FAS consists of nine questions and a scoring range of 0-27, with higher scores indicative of poorer bowel function. One hundred eighty-seven (98.4%) surveys were returned. The median follow-up was 66.3 months (range 54-92 months). Clinical, treatment, and dosimetric parameters evaluated for bowel dysfunction included patient age, diabetes, hypertension, tobacco consumption, clinical T stage, elapsed time since implant, prostate ultrasound volume, planning target volume, hormonal status, supplemental external beam radiation therapy (EBRT), isotope, average, median and maximum rectal doses, total implanted seed strength, values of the minimum dose received by 90% of the prostate gland (D(90)), and the percent prostate volume receiving 100%, 150%, and 200% of the minimum prescribed dose (V(100/150/200)). RESULTS: The R-FAS score for the evaluated cohort was 3.92, which represented a slight improvement from the 1999 survey score of 4.15 (p = 0.29). Of the evaluated clinical, treatment, and dosimetric parameters, the number of preimplant bowel movements, a history of tobacco consumption, and the median rectal dose correlated with the R-FAS. Although lower rectal doses were noted with Pd-103, isotope did not predict for bowel function. Only 12% (23/187) of patients reported their bowel function to be worse after implantation. Patient perception of overall rectal quality of life was inversely related to the use of supplemental EBRT (p = 0.007). CONCLUSION: Minor bowel changes are noted following prostate brachytherapy. The vast majority of patients did not report any deterioration in bowel function. In addition, the R-FAS continue to improve with time. PMID- 12909215 TI - Intensity-modulated radiation therapy for head-and-neck cancer: the UCSF experience focusing on target volume delineation. AB - PURPOSE: To review the University of California-San Francisco (UCSF) experience of using intensity-modulated radiation therapy (IMRT) to treat head-and-neck cancer focusing on the importance of target volume delineation and adequate target volume coverage. METHODS AND MATERIALS: Between April 1995 and January 2002, 150 histologically confirmed patients underwent IMRT for their head-and neck cancer at our institution. Sites included were nasopharynx 86, oropharynx 22, paranasal sinus 22, thyroid 6, oral tongue 3, nasal cavity 2, salivary 2, larynx 2, hypopharynx 1, lacrimal gland 1, skin 1, temporal bone 1, and trachea 1. One hundred seven patients were treated definitively with IMRT +/- concurrent platinum chemotherapy (92/107), whereas 43 patients underwent gross surgical resection followed by postoperative IMRT +/- concurrent platinum chemotherapy (15/43). IMRT was delivered using three different techniques: 1) manually cut partial transmission blocks, 2) computer-controlled auto-sequencing segmental multileaf collimator, and 3) sequential tomotherapy using dynamic multivane intensity-modulating collimator. Forty-two patients were treated with a forward plan, 102 patients with an inverse plan, and 6 patients with both forward and inverse plans. The gross target volume (GTV) was defined as tumor detected on physical examination or imaging studies. In postoperative cases, the GTV was defined as the preoperative gross tumor volume. The clinical target volume (CTV) included all potential areas at risk for microscopic tumor involvement by either direct extension or nodal spread including a margin for patient motion and setup errors. The average prescription doses to the GTV were 70 Gy and 66 Gy for the primary and the postoperative cases, respectively. The site of recurrence was determined by the diagnostic neuroradiologist to be either within the GTV or the CTV volume by comparison of the treatment planning computed tomography with posttreatment imaging studies. RESULTS: For the primary definitive cases with a median follow-up of 25 months (range 6 to 78 months), 4 patients failed in the GTV. The 2- and 3-year local freedom from progression (LFFP) rates were 97% and 95%. With a median follow-up of 17 months (range 8 to 56 months), 7 patients failed in the postoperative setting. The 2-year LFFP rate was 83%. For the primary group, the average maximum, mean, and minimum doses delivered were 80 Gy, 74 Gy, 56 Gy to the GTV, and 80 Gy, 69 Gy, 33 Gy to the CTV. An average of only 3% of the GTV and 3% of the CTV received less than 95% of the prescribed dose. For the postoperative group, the average maximum, mean, and minimum doses delivered were 79 Gy, 71 Gy, 37 Gy to the GTV and 79 Gy, 66 Gy, 21 Gy to the CTV. An average of only 6% of the GTV and 6% of the CTV received less than 95% of the prescribed dose. CONCLUSION: Accurate target volume delineation in IMRT treatment for head-and-neck cancer is essential. Our multidisciplinary approach in target volume definition resulted in few recurrences with excellent LFFP rates and no marginal failures. Higher treatment failure rates were noted in the postoperative setting in which lower doses were prescribed. Potential dose escalation studies may further improve the local control rates in the postoperative setting. PMID- 12909216 TI - Quality of life after parotid-sparing IMRT for head-and-neck cancer: a prospective longitudinal study. AB - PURPOSE: Parotid-sparing intensity-modulated radiotherapy (IMRT) for head-and neck cancer reduces xerostomia compared with standard RT. To assess potential improvements in broader aspects of quality of life (QOL), we initiated a study of patient-reported QOL and its predictors after IMRT. MATERIALS AND METHODS: This was a prospective longitudinal study of head-and-neck cancer patients receiving multisegmental static IMRT. Patients were given a validated xerostomia questionnaire (XQ), and a validated head-and-neck cancer-related QOL questionnaire consisting of four multi-item domains: Eating, Communication, Pain, and Emotion. The Eating domain contains one question (total of six) asking directly about xerostomia. In both questionnaires, higher scores denote worse symptoms or QOL. The questionnaires and measurements of salivary output from the major glands were completed before RT started (pre-RT) and at 3, 6, and 12 months after RT. The association between the QOL scores and patient-, tumor-, and therapy-related factors was assessed using the random effects model. RESULTS: Thirty-six patients participating in the study completed the questionnaires through 12 months. The XQ scores worsened significantly at 3 months compared with the pre-RT scores, but later they improved gradually through 12 months (p = 0.003), in parallel with an increase in the salivary output from the spared salivary glands. The QOL summary scores were stable between the baseline (pre-RT) and 3 months after RT scores. Patients receiving postoperative RT (whose pre-RT questionnaires were taken a few weeks after surgery) tended to have improved scores after RT, reflecting the subsidence of acute postoperative sequelae, compared with a tendency toward worsened scores in patients receiving definitive RT. After 3 months, statistically significant improvement was noted in the summary QOL scores for all patients, through 12 months after RT (p = 0.01). The salivary flow rates, tumor doses, mean oral cavity dose, age, gender, sites or stages of tumor, surgery, and use of chemotherapy were not associated with the QOL scores at any point. The mean dose to the parotid glands correlated with the QOL scores at 3 months (p = 0.05) but not at other post-RT periods. The XQ and QOL summary scores did not correlate before RT but were significantly correlated at each post-RT point (p < 0.01), with a linear correlation coefficient (r) of 0.59, 0.72, and 0.67 at 3, 6, and 12 months, respectively. At these points, the XQ scores also correlated significantly with the scores of each of the individual QOL domains (p < or = 0.01), including the domains Pain and Emotion, which did not contain any xerostomia-related question. CONCLUSION: After parotid-sparing IMRT, a statistically significant correlation was noted between patient-reported xerostomia and each of the domains of QOL: Eating, Communication, Pain, and Emotion. Both xerostomia and QOL scores improved significantly over time during the first year after therapy. These results suggest that the efforts to improve xerostomia using IMRT may yield improvements in broad aspects of QOL. PMID- 12909217 TI - Effect of treatment time on outcome of radiotherapy for oral tongue carcinoma. AB - PURPOSE: To investigate the importance of total treatment time on the outcome of external beam radiotherapy (EBRT) followed by internal brachytherapy for the treatment of oral tongue carcinoma. METHODS AND MATERIALS: Ninety-four patients with T1-T2N0 squamous cell carcinoma of the oral tongue were treated using 35-40 Gy EBRT followed by 35-40 Gy interstitial (137)Cs brachytherapy between 1985 and 1995. The interval between the end of EBRT and the start of interstitial treatment varied for numerous unavoidable reasons, with a mean of 25.3 days and standard deviation of 3.5 days. The median follow-up period was 59.1 months (range 6-146). RESULTS: The actuarial survival rate of all cases was 78.4% at 5 years. The 5-year local control rate for those with T1 and T2 was 92.8% and 62.7%, respectively (p < 0.05). The local control rate of the primary tumor in patients with a total treatment time >43 days was statistically lower than that of patients with a total treatment time < or =43 days in all patients (p < 0.05) and in the subgroup of Stage T2 patients (p < 0.05). Multivariate analysis revealed that the local control rates in all cases were significantly related to the T stage (T2 or not), total treatment time (>43 days or not), and location of disease (posterior or not). Regression analysis for 5-year local control as a function of treatment duration showed a 2% loss of local control per day of treatment extension >30 days (r = 0.94, p < 0.01). CONCLUSION: The total treatment time was associated with the local control rate in the RT of oral tongue carcinoma. The loss in local control was estimated to be 2.0% per additional day in our series for oral tongue carcinoma. PMID- 12909218 TI - Oropharyngeal candidiasis caused by non-albicans yeast in patients receiving external beam radiotherapy for head-and-neck cancer. AB - PURPOSE: To characterize non-albicans Candida oral infections in patients with head-and-neck cancer receiving external beam radiotherapy (EBRT) with or without concurrent chemotherapy. METHODS AND MATERIALS: Thirty-seven patients with head and-neck cancer received EBRT in 2.0-Gy daily fractions to a median dose of 60.4 Gy (range 38-82.8, mean 64.6). They were followed for oropharyngeal candidiasis (OPC) confirmed by positive examination, positive KOH smear, and/or positive swab or swish culture. Samples were identified and plated on chromogenic media to identify non-albicans yeasts. Colonies were plated on Sabouraud dextrose slants for microdilution antifungal susceptibility testing to fluconazole. DNA typing, including karyotyping, restriction fragment length polymorphism analysis, and Southern blot hybridization with the moderately repetitive Ca3 probe, was performed on selected isolates to confirm individual species. RESULTS: Of the 37 patients, 10 (27%) developed OPC, and 26 (70.3%) displayed Candida carriage state. The median EBRT dose at time of positive culture was 22.5 Gy and at time of OPC was 28.6 Gy. Of the 6 patients receiving chemotherapy and EBRT, 4 (66%) developed OPC at median dose of 27.6 Gy. Three (8%) of 37 patients were infected with non-albicans Candida, and 3 (30%) of all 10 infections were caused by these organisms. CONCLUSION: Non-albicans Candida is emerging as a relatively common cause of OPC in head-and-neck cancer patients. Chromogenic media are helpful to screen these infections. Our data also suggest a greater likelihood of developing OPC in patients receiving concomitant chemotherapy and EBRT. PMID- 12909219 TI - The addition of continuous infusion 5-FU to preoperative radiation therapy increases tumor response, leading to increased sphincter preservation in locally advanced rectal cancer. AB - PURPOSE: To compare the outcome from preoperative chemoradiation (CXRT) and from radiation therapy (RT) in the treatment of rectal cancer in two large, single institutional experiences. PATIENTS AND METHODS: Between 1978 and 1995, 403 patients with localized, nonmetastatic, clinically staged T3 or T4 rectal cancer patients were treated with preoperative RT alone at two institutions. Patients at institution 1 (n = 207) were treated with pelvic CXRT exclusively, and patients at institution 2 were treated (except for 8 given CXRT) with pelvic RT alone (n = 196). In addition, a third group (n = 61) was treated with CXRT at institution 2 between 1998 and 2000 after a policy change. Both institutions delivered 45 Gy in five fractions as a standard dose, but institution 2 used 20 Gy in five fractions in selected cases (n = 26). At both institutions, concurrent chemotherapy consisted of a continuous infusion of 5-fluorouracil (5-FU) at a dosage of 1500 mg/m(2)/week. The end points were response, sphincter preservation (SP), relapse free survival (RFS), pelvic disease control (PC), and overall survival (OS). RESULTS: Median follow-up was 63 months for all living patients at institution 1 and in the primary group of institution 2. Multivariate analysis of the patients in these groups showed that the use of concurrent chemotherapy improved tumor response (T-stage downstaging, 62% vs. 42%, p = 0.001, and pathologic complete response, 23% vs. 5% p < 0.0001), but did not significantly improve LC, RFS, or OS. Follow-up for the secondary group at institution 2 was insufficient to allow the analysis of these endpoints. In the subset of patients receiving 45 Gy who had rectal tumors < or /=6 cm from the anal verge (institution 1: n = 132; institution 2 primary: n = 79; institution 2 secondary: n = 33), there was a significant improvement in SP with the use of concurrent chemotherapy (39% at institution 1 compared with 13% in the primary group at institution 2, p < 0.0001). A logistic regression analysis of clinical prognostic factors indicated that the use of concurrent chemotherapy independently influenced SP in these low tumors (p = 0.002). This finding was supported by a 36% SP rate in the secondary group at institution 2. Thus SP increased after the addition of chemotherapy at institution 2. CONCLUSIONS: The use of concurrent 5-FU with preoperative radiation therapy for T3 and T4 rectal cancer independently increases tumor response and may contribute to increased SP in patients with low rectal cancer. PMID- 12909220 TI - Predictive value of carbohydrate antigen 19-9 in pancreatic cancer treated with radiochemotherapy. AB - PURPOSE: To determine the predictive value of carbohydrate antigen (CA) 19-9 in pancreatic cancer treated with radiochemotherapy. METHODS AND MATERIALS: Ninety five patients with locally advanced unresectable adenocarcinoma of the pancreas were treated with hyperfractionated accelerated radiotherapy to a total dose of 44.8 Gy combined with 5-fluorouracil and folinic acid. CA 19-9 was measured before therapy, each week during therapy, and every 4 weeks during the follow-up period. RESULTS: The median CA 19-9 before treatment was 420 U/mL; in the responder group it was 117 U/mL, and in the nonresponder group it was 806 U/mL. Patients with a pretreatment CA 19-9 less than the median had not only a significantly better tumor response (45.8%) but also a better survival prognosis (median survival 12.3 months) than those with a level higher than the median (tumor response 12.8%; median survival 7.1 months). The posttreatment median CA 19-9 for all patients also exhibited prognostic significance. The median survival of patients with a CA 19-9 level lower than the posttreatment median of 293 U/mL was 13.5 months, compared with 7.2 months for those with a CA 19-9 level greater than the median. To detect recurrent disease during follow-up, the sensitivity of CA 19-9 was 100% and the specificity 88%. CONCLUSION: Our results indicate that CA 19-9 is of predictive value for prognosis, response, and detecting recurrence of pancreatic cancer in patients undergoing combined radiochemotherapy. Therefore, we recommend the routine implementation of CA 19-9 observation during the clinical course of treatment for patients with pancreatic cancer undergoing radiochemotherapy. PMID- 12909221 TI - Concurrent chemoradiotherapy treatment of locally advanced pancreatic cancer: gemcitabine versus 5-fluorouracil, a randomized controlled study. AB - PURPOSE: To determine the efficacy and tolerability of gemcitabine (GEM) concurrent chemoradiotherapy (CCRT) vs. 5-fluorouracil (5-FU) CCRT for locally advanced pancreatic cancer. METHODS AND MATERIALS: Thirty-four patients with locally advanced pancreatic cancer were studied. Eighteen patients were randomized to receive GEM CCRT (600 mg/m(2)/wk for 6 weeks) and 16 patients to receive bolus 5-FU CCRT (500 mg/m(2)/d for 3 days repeated every 2 weeks for 6 weeks). All patients were to receive 3D-CRT 50.4-61.2 Gy at 1.8-Gy/d fractions and GEM (1000 mg/m(2) weekly for 3 weeks repeated every 4 weeks) after RT. RESULTS: The median survival and median time to progression were 14.5 months and 7.1 months for the GEM CCRT group and 6.7 months and 2.7 months for the 5-FU CCRT group (p = 0.027 and p = 0.019, respectively). The quality-adjusted life month survival time was 11.2 +/- 0.5 months for GEM CCRT and 6.0 +/- 0.3 months for 5 FU CCRT patients (p <0.001). The response rate was 50% (four complete responses and five partial responses) for GEM CCRT and 13% (two partial responses) for 5-FU CCRT (p = 0.005). Pain control was 39% for GEM CCRT and 6% for 5-FU CCRT (p = 0.043). Grade 3-4 neutropenia (34% vs. 19%), thrombocytopenia (0% vs. 7%), nausea (33% vs. 31%), vomiting (17% vs. 19%), hospitalization days per month of survival (7.4 +/- 1.7 days vs. 8.0 +/- 1.3 days), and full dose of RT received (78% vs. 75%) were not significantly different between the GEM CCRT and 5-FU CCRT patients. CONCLUSION: GEM CCRT appears more effective than 5-FU CCRT for locally advanced pancreatic cancer and has comparable tolerability. PMID- 12909222 TI - Combination of external beam irradiation and high-dose-rate intraluminal brachytherapy for inoperable carcinoma of the extrahepatic bile ducts. AB - PURPOSE: To assess the feasibility and therapeutic benefits of a combination of external beam radiotherapy (EBRT) and high-dose-rate intraluminal brachytherapy (ILBT) for treating patients with inoperable carcinoma of the extrahepatic bile ducts. METHODS AND MATERIALS: Of 31 patients who received RT at the Yonsei Cancer Center, Yonsei University College of Medicine in Seoul, Korea between 1986 and 1995, 17 patients underwent EBRT alone (Group 1) and 14 patients were treated with EBRT in combination with high-dose-rate ILBT (Group 2). After external drainage, EBRT was delivered with a total dose ranging from 36 to 55 Gy (median 50.4) in both groups. High-dose-rate ILBT for the patients in Group 2 was performed using a high-intensity (192)Ir source (Gamma-med remote afterloading system) within the expandable intrabiliary prosthesis (Gianturco stent), inserted transhepatically at the site of the obstruction. The radiation dose of the high dose-rate ILBT was prescribed at 1.5 cm from the center of the source with a single daily dose of 5 Gy to a total of 15 Gy given in three fractions. The response rate, patterns of treatment failure, treatment morbidity, and survival data in the two groups were compared. RESULTS: Although locoregional recurrence was the most common pattern of failure in both groups, no statistically significant difference was found in the recurrence rates between those who did and did not receive ILBT (53% for Group 1 vs. 36% for Group 2; p > 0.05). However, a prolongation of the median time to tumor recurrence was observed in the Group 2 patients (5 months for Group 1 vs. 9 months for Group 2; p = 0.06). When the EBRT dose delivered was >50 Gy, most patients experienced various degrees of GI symptoms, but the frequency of radiation-induced complications in the two groups was similar. No enhancement in treatment morbidity was attributed to the addition of high-dose-rate ILBT to EBRT. With a median follow-up of 12 months, the overall actuarial 2-year survival rate for Group 2 patients was significantly better than that for Group 1 patients (0% for Group 1 vs. 21% for Group 2; p = 0.015). CONCLUSION: Given these observations, we believe that the combined use of EBRT and high-dose-rate ILBT is a beneficial, relatively safe, and effective method of improving the treatment outcome in selected patients with inoperable carcinoma of the extrahepatic bile ducts. PMID- 12909223 TI - Prospective trial of combined transcatheter arterial chemoembolization and three dimensional conformal radiotherapy for portal vein tumor thrombus in patients with unresectable hepatocellular carcinoma. AB - PURPOSE: We conducted a prospective trial of combined transarterial chemoembolization (TACE) and three-dimensional conformal radiotherapy (3D-CRT) for portal vein tumor thrombus (PVTT) in unresectable hepatocellular carcinoma (HCC). The aim of the present study was to investigate the efficacy and toxicity of this trial regime. METHODS AND MATERIALS: Patients with unresectable HCC complicated with tumor thrombus in the first branch of portal vein were selected as eligible for this study. TACE was performed using Lipiodol, epirubicin hydrochloride, and mitomycin, followed by gelatin sponge cubes. The 3D-CRT was performed targeting the clinical target volume (CTV) defined as PVTT only to a total dose of 60 Gy using 10-MV accelerator. RESULTS: A total of 19 patients were enrolled in this study. Survival rates at 1 and 2 years were 40.6% and 10.2%, respectively. The median survival time was 7.0 months. An objective response was observed in 11 of 19 cases (57.9%). Recanalization of the first portal branches was not observed; however, the protrusion of PVTT into the main portal trunk decreased in all cases. Growth of intrahepatic metastasis outside the 3D-CRT field was observed in 12 cases (63%). Deterioration of the Child-Pugh Score was observed in 5 of 6 cases with the percent volume of the total liver receiving a dose exceeding 30 Gy (V(30)) > or =40%, vs. 2 of 13 cases with a V(30) <40% (p < 0.01). CONCLUSION: This combined therapy was feasible. Our results indicate that V(30) was a predictive test for deterioration of liver function. Further investigation of treatment modalities is needed to prevent the growth of intrahepatic metastasis. PMID- 12909224 TI - Long-term outcome of phase II trial evaluating chemotherapy, chemoradiotherapy, and surgery for locoregionally advanced esophageal cancer. AB - PURPOSE: To evaluate the long-term outcome of chemotherapy, chemoradiotherapy, and surgery for patients with locoregionally advanced esophageal cancer. METHODS AND MATERIALS: Thirty-eight patients with locoregionally advanced esophageal cancer were entered into a Phase II study between November 1996 and October 1998 at the University of Texas M. D. Anderson Cancer Center. Patients initially received two cycles of chemotherapy with paclitaxel (200 mg/m(2)), 5-fluorouracil (750 mg/m(2)/d for 5 days), and cisplatin (15 mg/m(2)/d for 5 days), followed by chemoradiotherapy, consisting of radiation (45 Gy during 5 weeks) with 5 fluorouracil (300 mg/m(2)/d during radiation) and cisplatin (15 mg/m(2)/d for 5 days). Surgical resection was performed 4-6 weeks after the completion of the chemoradiotherapy. RESULTS: Most patients had adenocarcinoma (n = 32; 84%). Pretreatment endoscopic ultrasonography revealed T3 tumors in 33 patients (87%) and N1 disease in 25 patients (66%). Thirty-seven patients (97%) completed the planned chemotherapy and chemoradiotherapy, and 35 patients (92%) underwent surgery, with a 30-day mortality rate of 6% (2 of 35 patients). A pathologic complete response or microscopic residual carcinoma (<10% viable) was found in 25 (71%) of 35 patients and was associated with a disease-free survival rate of 72% at 3 years and 51% at 5 years. On the basis of an intention-to-treat analysis and a median potential follow-up of 58 months, the 3- and 5-year overall survival rate for all 38 patients was 63% and 39%, respectively. CONCLUSION: The long-term results of this study suggest that the strategy of induction chemotherapy followed by chemoradiotherapy and surgery is safe and warrants further evaluation in the treatment of patients with locoregionally advanced esophageal cancer. PMID- 12909225 TI - Outcome of treatment for breast cancer patients with chest wall recurrence according to initial stage: implications for post-mastectomy radiation therapy. AB - PURPOSE: The impact of postmastectomy radiation therapy (PMRT) on overall survival (OS) for patients with Stage II breast cancer with 1-3 positive lymph nodes is controversial. We sought to compare the outcome of salvage treatment for patients with chest wall recurrence (CWR) according to initial disease stage to shed light on the potential benefit of PMRT in specific subgroups of patients. METHODS: We retrospectively reviewed information concerning 96 patients with CWR who were not previously treated with PMRT. The patients were divided according to their initial extent of disease: T1-T2N0 (Group 1), T1-T2 with 1-3 positive lymph nodes (Group 2), and T3-T4 or > or =4 positive lymph nodes (Group 3). The OS and distant metastasis-free survival (DMFS) from the time of CWR were compared using the method of Kaplan and Meier, and a Cox regression model was used for a multivariate analysis. RESULTS: Group 1 had an improved OS and DMFS compared with Group 2 and Group 3 (p < 0.001), but there were no differences in OS or DMFS between Group 2 and Group 3 (p = 0.250 and p = 0.492, respectively). The respective 5-year rates for the three groups were as follows: OS 79.9% vs. 41.9% vs. 29.1%; DMFS 75.2% vs. 33.6% vs. 25.9%. CONCLUSIONS: Breast cancer patients with T1-T2N0 breast cancer who develop a CWR have a significantly better outcome than those with lymph node-positive disease. Patients with T1-T2 tumors and one to three positive lymph nodes have a similar outcome after CWR as those with larger tumors or more than four positive lymph nodes. These data should be considered when weighing the risks and benefits of PMRT for patients with Stage II breast cancer with one to three positive lymph nodes. PMID- 12909226 TI - Radiotherapy and immediate two-stage breast reconstruction with a tissue expander and implant: complications and esthetic results. AB - PURPOSE: To report complications, failure rate, and esthetic results in patients undergoing immediate breast reconstruction with a tissue expander and implant, with or without adjuvant treatment. METHODS AND MATERIALS: We reviewed the records of the 77 patients who underwent immediate breast reconstruction with an expander/implant between January 1999 and December 2000. Complications were assessed using the Common Toxicity Criteria, version 2, scale. Esthetic results were assessed by the physician using five criteria. RESULTS: Of the 77 patients, 55 had received adjuvant radiotherapy. The median follow-up was 25 months. Complications appeared to correlate with radiotherapy (14% for nonirradiated patients; 51% for irradiated patients; p = 0.006) and adjuvant chemotherapy (54% with chemotherapy [CHT] vs. 25% without CHT; p = 0.02). Breast reconstruction failed in 21% of patients (9% of nonirradiated patients and 24% of irradiated patients; p = 0.1), and chemotherapy was associated with a worse rate of failure (34% with CHT vs. 6% without CHT, p = 0.005). Adjuvant tamoxifen, however, correlated neither with complications (45% with tamoxifen vs. 39% without; p = 0.15) nor with failure (21% with tamoxifen and 23% without, p = 0.79). Esthetic results were acceptable in 60% of cases. CONCLUSION: Immediate breast reconstruction with an expander/implant can be considered even for patients requiring adjuvant treatment. However, the complication and failure rates are three times higher after postexpander radiotherapy. PMID- 12909227 TI - Radiotherapy in conjunction with intravenous infusion of 180 mg of disodium pamidronate in management of osteolytic metastases from breast cancer: clinical evaluation, biochemical markers, quality of life, and monitoring of recalcification using assessments of gray-level histogram in plain radiographs. AB - PURPOSE: To evaluate the clinical improvement and radiographically monitor the effect of local radiotherapy in conjunction with disodium pamidronate (DP) on metastatic osteolytic disease. METHODS AND MATERIALS: Thirty-three patients with osteolytic metastasis from advanced breast cancer received radiotherapy with a 6 MV linear accelerator up to a dose of 30 Gy (3 Gy/fraction, 5 d/wk) combined with 24 monthly sessions of a 180-mg DP infusion. Conventional X-rays were obtained during the first six sessions of DP treatment, retaining the same settings for each exposure. The analysis of the image attributes was based on measuring the first-order statistics of the mean value and energy of gray-level histograms in the osteolytic region. RESULTS: The 6-month measurements compared with baseline showed statistically significant differences (p < 0.01, Wilcoxon test) in energy of gray-level histogram (-10.8%), mean value of gray-level histogram (+9.5%), pain score (-5.8 points), Eastern Cooperative Oncology Group status (-2.4 points), urine hydroxyproline/creatinine ratio (-41.7%), urine calcium/creatinine ratio (-58.8%), and bone alkaline phosphatase (-42.4%). Quality of life as determined by the European Organization for Research and Treatment of Cancer Core Quality of Life Questionnaire (version 3) also improved. During follow-up, 88% of patients had complete and 12% partial responses (International Union Against Cancer radiologic criteria). On multivariate analysis, greater changes in the mean value of the gray-level histogram and negative nodal status were predictors for a reduction in the number of skeletal complications after therapy. Flu-like syndrome occurred in 13 patients (39%) and was well managed with mild antipyretics. CONCLUSION: Image-processing in plain radiographs offers an objective way to assess recalcification. The image-processing indexes, along with the measurements of performance status, quality of life, and biochemical markers, improved significantly. Local radiotherapy combined with long-term high-dose DP up to 180 mg is tolerable and has a high therapeutic response. PMID- 12909228 TI - Radiotherapy in the management of giant cell tumor of bone. AB - PURPOSE: To evaluate the outcomes of patients with giant cell tumor of bone (GCTB) treated with radiotherapy (RT) with or without surgical resection. METHODS AND MATERIALS: We performed a retrospective review of the records from 25 consecutive patients with pathologically confirmed GCTB who had undergone RT between 1956 and 2000. RESULTS: Patients ranged in age from 11 to 69 years (median 32); 16 were female and 9 were male. The anatomic distribution of lesions was as follows: cervical spine, 3; temporal bone, 1; thoracic or lumbar spine, 9; sacrum, 8; ilium, 1, and humerus, radius, and thumb metacarpal, 1 each. Tumors ranged in size from 2 to 20 cm (median 9.5) at their maximal dimension. Thirteen patients had been referred for RT for primary GCTB and 12 had been referred with locally recurrent disease after having undergone one or more other treatments. Fourteen patients had undergone RT for gross disease, and the remaining 11 had been treated with RT after gross total resection. In 10 of these 11 patients, the treatment margins were positive or uncertain. Radiation doses ranged from 25 to 65 Gy (median 46). At a median follow-up of 8.8 years (range 0.67-34), 7 patients had developed isolated local recurrence, 2 had developed isolated distant recurrence, and 3 had developed both. The actuarial 5-year overall and disease free survival rate was 91% and 58%, respectively, and the actuarial 5-year local control and distant metastasis-free survival rate was 62% and 81%, respectively. Univariate analysis suggested that treatment for recurrent disease correlated with a lower disease-free survival rate (83% vs. 33%, p = 0.06), distant metastasis-free survival rate (100% vs. 64%, p = 0.08), and local control rate (83% vs. 42%, p = 0.08) at 5 years. Of the 12 cases of recurrence, 7 were ultimately successfully treated with additional salvage therapy. In 4 of these patients, salvage therapy included interferon-alpha 2b. CONCLUSION: RT should be considered an adjuvant to surgery or as alternative therapy in cases of GCTB that are unresectable or in which excision would result in substantial functional deficits. When RT is used as primary therapy, the rate of local control seems to be satisfactory. In heavily pretreated patients, however, RT delivered as it was in this series can result in poor local control, and alternative therapies should be considered. PMID- 12909229 TI - Phase I study of escalating doses of low-dose-rate, locoregional irradiation preceding Cytoxan-TBI for patients with chemotherapy-resistant non-Hodgkin's or Hodgkin's lymphoma. AB - PURPOSE: In patients in whom bone marrow transplantation (BMT) fails, recurrence often occurs at sites known to have contained disease before initiating BMT. The purpose of this study was to find the maximal tolerable dose of locoregional irradiation (LRT) between 1000 and 2000 cGy that could be integrated with our Cytoxan-total body irradiation (TBI) BMT conditioning regimen in the treatment of lymphoma. METHODS AND MATERIALS: Patients had Hodgkin's or non-Hodgkin's lymphoma in chemotherapy-refractory relapse. All patients received LRT to a maximum of three sets of fields encompassing either all current or all previously known sites of disease. Cytoxan-TBI consisted of cyclophosphamide 50 mg/kg daily for 4 days followed by TBI of 1200 cGy given in four fractions. RESULTS: Twenty-one patients were enrolled. Radiation Therapy Oncology Group Grade 3 in-field acute toxicity was observed in 1 patient at each dose level up to 1500 cGy and in 3 of 6 patients receiving 2000 cGy. Clinically evident late toxicities were limited to hypothyroidism and one second malignancy occurring outside the LRT fields. CONCLUSION: Low-dose-rate, LRT with concurrent Cytoxan-TBI before BMT has acceptable rates of in-field toxicity for doses up to 1500 cGy in five fractions. This regimen safely permits the use of a total combined radiation dose of up to 2700 cGy during 2 weeks, with encouraging in-field response rates in treatment refractory patients. PMID- 12909230 TI - Treatment results of radiotherapy for malignant lymphoma of the orbit and histopathologic review according to the WHO classification. AB - PURPOSE: To analyze the results of radiotherapy (RT) for malignant lymphoma of the orbit and to evaluate them compared with the World Health Organization (WHO) classification published in 2001. METHODS AND MATERIALS: The data from 29 patients with malignant lymphoma of the orbit treated with RT at Gunma University Hospital between 1978 and 2001 were retrospectively analyzed. Pathologic slides from 23 cases were available and were reviewed by a hematopathologist according to the WHO classification. The original and reviewed diagnoses, patient characteristics, treatment results, and complications were analyzed. In principle, patients with low-grade or indolent lymphoma were treated with RT alone, using 30 Gy as the tumor dose. Survival data were calculated using the Kaplan-Meier method. RESULTS: One case that proved to be a pseudotumor was excluded from evaluation. Of the 28 cases, 25 were Stage IAE, 1 was Stage IIAE, and 2 were Stage IVAE. The median follow-up was 71 months. According to the original classification and the Working Formulation, the 5- and 10-year overall survival rate of patients with low-grade lymphoma was 94% and 73%, respectively. The corresponding rates for those with intermediate-grade lymphoma were 67% and 67% (p = 0.15). In contrast, the WHO classification showed a significant difference in the survival curves. The 5- and 10-year overall survival rate of patients with mucosa-associated lymphoid tissue (MALT) lymphoma was 100% and 88%, respectively; for diffuse large B-cell patients, the rates were both 0% (p < 0.001). In patients with MALT lymphoma, one local and four distant relapses developed; two of them >10 years after initial treatment. All of the relapsed MALT lymphomas were controlled by salvage therapy. CONCLUSION: Excellent local control and survival can be achieved for patients with orbital MALT lymphoma using RT alone. A precise histopathologic diagnosis using the WHO classification and long-term follow-up for >10 years is recommended. PMID- 12909231 TI - Brain metastasis in children with sarcoma, neuroblastoma, and Wilms' tumor. AB - PURPOSE: To determine the incidence and prognosis of children who develop brain metastasis. MATERIALS AND METHODS: The medical and tumor registry records of 611 children treated at the University of Iowa Hospitals and Clinics between 1965 and 2000 for a sarcoma, neuroblastoma, or Wilms' tumor were reviewed. RESULTS: Thirty children (4.9%) were found to have brain metastasis. Brain metastasis occurred in 9 of 113 (8%) neuroblastoma, 7 of 104 (6.7%) rhabdomyosarcoma, 6 of 105 (5.7%) Ewing's sarcoma, 5 of 106 (4.7%) osteosarcoma, 2 of 83 (2.4%) nonrhabdomyosarcoma soft-tissue sarcoma, and 1 of 100 (1%) Wilms' tumor patients. There were 22 male and 8 female patients, with a median age of 14 years at the time of diagnosis of brain metastasis (range 8 months-20 years). Four patients were diagnosed at autopsy, whereas five had brain metastasis at initial diagnosis. For the 25 children who did not have brain metastasis at initial presentation, the median interval from initial diagnosis to development of brain metastasis was 5 months (range 1-43 months). Twenty-nine (97%) had concurrent or prior history of distant metastasis. Eighteen (60%) had solitary brain metastasis. Treatment for the 26 non-autopsy-diagnosed children included surgery (S), followed by postoperative radiotherapy (RT) and chemotherapy (CT) in 2, S and postoperative RT in 1, RT and CT in 16, S alone in 2, CT alone in 2, RT alone in 1, and no treatment in 2. Median survival was 4 months after diagnosis of brain metastasis, with a 1-year survival rate of 11.5%. On multivariate analysis, only the use of RT was found to positively influence freedom from neurologic progression (p = 0.005). CONCLUSION: Brain metastasis is uncommon in children with a diagnosis of sarcoma, neuroblastoma, or Wilms' tumor and is often accompanied by concurrent distant disease. Children treated with RT for brain metastasis had a better freedom from neurologic progression rate than those who did not receive radiotherapy. PMID- 12909232 TI - Radiosurgery of cerebral arteriovenous malformations in children: a series of 57 cases. AB - PURPOSE: To evaluate the efficacy and outcome of Linac radiosurgery (RS) as treatment of cerebral arteriovenous malformations (cAVM) in a series of 57 children. METHODS AND MATERIALS: Between 1984 and 2000, we used Linac radiosurgery to treat 792 patients with cAVM. This series included 57 children (7.2%) under the age of 15 years at the time of RS (range = 7-15 years, median = 12 years). We were able to evaluate 49 of the children (86%) by angiography, 21 boys and 28 girls (sex ratio = 0.75). First symptoms were: hemorrhage, 34 patients (69.4%); seizures, 6 patients (12.5%); headache, 6 patients (12.5%); and progressive neurologic deficit, 1 patient (2.1%). Nidus size ranged from 5 to 50 mm (median = 20 mm). Nidus volume ranged from 0.6 to 16 cc (median = 3.5 cc). Patient distribution according to Spetzler-Martin grade was as follows: Grade 1, n = 5 (11%); Grade 2, n = 18 (35%); Grade 3, n = 21 (40%,); Grade 4, n = 5 (14%); and Grade 5, n = 0%. Twenty-seven patients (55.1%) had other treatment before RS: embolization, n = 14 (31.1%); neurosurgery, n = 9 (20.5%); embolization and neurosurgery, n = 3 (6.1%). RS was performed with the system used for adults. Patients were seated in a Betti armchair. Circular 15 MV X-ray minibeams (6 to 20 mm) were delivered in coronal arcs by a GECGR Saturne 43 Linac. Planification and dosimetry were carried out using the Associated Target Methodology and Dosigray TPS dosimetric systems. The dose at the peripheral isodose (50-70%) ranged from 18 to 28 Gy. Median and mean doses were 25 Gy and 23.8 Gy. Mono-isocentric planification was used in 25 patients (53.2%) and multi-isocentric in 24 patients (2 to 5 isocenters). The overall follow-up ranged from 7 to 172 months (mean 40 months, median 34 months). RESULTS: The overall rate of obliteration (OR) was 30/49 (61.2%). Mean time to obliteration was 34 months (range = 7 to 172 months). OR varied according to nidus size and volume: OR was 80% for nidus <15 mm, 67% for nidus between 15 and 25 mm, and 42% for nidus >25 mm (p = 0.058). OR was 100% for nidus <1 cc, 73% for nidus between 1 and 4 cc, and 40% for nidus of 4 to 10 cc (p = 0.019). OR according to patient gender was 84.2% for boys and 40% for girls. OR according to minimum dose (Dmin) was 44% for Dmin < 15 Gy (p = 0.01), 89% for D min from 15 to 20 Gy, and 100% for Dmin > 20 Gy (p = 0.01). OR was 62% in nonembolized AVM and 58% in previously embolized AVM (NS). OR according to the number of isocenters was 68.2%, 55.6%, 80%, 50%, and 0% for 1, 2, 3, 4, and 5 isocenters, respectively. After multivariate analysis, only Dmin closely correlated with OR (beta = 0.462; SE = 0.244, p = 0.057). Of the 6 patients with seizures before RS, 5 (80%) were seizure-free without medication after RS. One patient died of pneumonia. MORBIDITY: Four patients (8.2%) had bleeding after RS at 39, 45, 51, and 59 months. No new neurologic deficit was found during the follow-up period. Twenty-nine patients underwent magnetic resonance imaging. Thirteen patients (44.8%) showed no parenchymal changes. Thirteen patients (44.8%) had Grade 2 changes, that is, T2 hypersignals. Two patients (6.9%) had Grade 3 changes, and 1 patient (3.4%) had Grade 4 "necrosis-like" changes. CONCLUSION: In our experience, Linac RS has proven to be a safe and effective method to treat cerebral AVM in children <15 years, whether used alone or in association with embolization and/or neurosurgery. PMID- 12909233 TI - Radiation tolerance of normal temporal bone structures: implications for gamma knife stereotactic radiosurgery. AB - Popular current thought states that hearing loss and facial weakness after radiosurgery of vestibular schwannomas is a function of cranial nerve damage. Although this may be true in some cases, the middle and inner ear contain rich networks of other sensitive structures that are at risk after radiotherapy and that may contribute to toxicity afterward. We reviewed the limited reported data regarding radiation tolerance of external, middle, and inner ear structures, and perspectives for therapy with gamma knife stereotactic radiosurgery are addressed. PMID- 12909234 TI - High-dose whole abdominal and pelvic irradiation for treatment of ovarian carcinoma: long-term toxicity and outcomes. AB - PURPOSE: To evaluate the role of high-dose whole abdominal and pelvic irradiation (WART) in the treatment of epithelial ovarian carcinoma. METHODS AND MATERIALS: A retrospective review was performed on 71 patients with Stage I-III ovarian carcinoma who were treated with WART using an open field technique after total abdominal hysterectomy and bilateral oophorectomy with or without omentectomy. Whole abdominal doses greater than typically recommended were used in a series of patients to enhance local control and to decrease abdominal recurrence. None of the patients had received chemotherapy before RT. Thirty-one patients received Alkeran or cyclophosphamide and two received cisplatin-based chemotherapy after WART. The median whole abdominal dose was 36 Gy (range 9-45.5), delivered in a median of 30 fractions (range 8-46). A pelvic boost was delivered using AP-PA fields during whole abdominal RT to a total midline pelvic dose of 200 cGy/d. The median pelvic dose was 51 Gy (range 16-59). The right lobe and a portion of the left lobe of the liver were shielded with custom blocks at a median dose of 25 Gy (range 9-41). The kidneys were shielded either AP-PA or PA from the first day of RT. The median dose to the kidneys was 19 Gy (range 4-30). RESULTS: The 5-year overall survival rate was 93%, 48%, and 29% for Stage I, II, and III patients, respectively. On multivariate analysis, stage and the extent of residual disease were independent prognostic factors. The 5- and 10-year overall survival rate for the 46 patients in the intermediate-risk group was 61% and 54%, respectively. For this group, a total abdominal dose of > or /=36 Gy was associated with a longer overall survival independent of stage, grade, and the amount of residual disease. This was most likely due to a significant reduction in the incidence of abdominal recurrence in patients receiving >36 Gy to the whole abdomen (18% vs. 49%, p = 0.006). Multivariate analysis revealed that grade (p = 0.023) and abdominal dose (p = 0.018) were independent factors influencing the rate of abdominal recurrence. Pelvic recurrence was rare as a first site of failure (6%). Twenty one percent (n = 15) of the patients developed Grade 3 or 4 (Radiation Therapy Oncology Group [RTOG] criteria) chronic small or large bowel toxicity. Eleven percent of all patients had a small bowel obstruction requiring surgery. A whole abdominal dose >30 Gy and a pelvic dose >50 Gy were associated with a significant increase in small bowel obstruction (p = 0.01) independent of other factors. Chronic Grade 3 or 4 (Common Toxicity Criteria) anemia, thrombocytopenia, and leukopenia were seen in 7%, 1%, and 4% of the patients, respectively. Transient liver enzyme elevation was common (62%). Two patients had Grade 3 (RTOG) hepatic toxicity. Grade 3 or 4 renal toxicity (RTOG) was observed in 4%, and 2 patients (3%) were diagnosed with pelvic insufficiency fractures that were managed conservatively. CONCLUSION: Survival after RT for ovarian carcinoma rivals that achieved with systemic chemotherapy. The results of this study suggest a possible dose-control relationship between the whole abdominal dose and the risk of abdominal recurrence; however, a higher rate of small bowel obstruction was observed when greater abdominal doses and greater pelvic doses were combined. Careful attention to balancing toxicity and efficacy is imperative if RT is to have a future role in the treatment of this disease. PMID- 12909235 TI - Patterns of failure after the multimodality treatment of uterine papillary serous carcinoma. AB - PURPOSE: Uterine papillary serous carcinoma (UPSC) is an aggressive variant of endometrial carcinoma. The majority of patients with clinical Stage I UPSC are found to have extrauterine disease at the time of surgery. Most authors report survival rates of 35-50% for Stage I-II and 0-15% for Stage III and IV UPSC. Surgical treatment as the sole therapy for patients with Stage I-IV UPSC is unacceptable because of high recurrence rates. Chemotherapy, radiotherapy, or both have been added after surgery in an attempt to improve survival. However, the survival benefit to patients from such multimodality therapy remains uncertain. This study analyzes the patterns of failure in patients with FIGO Stages I-IV UPSC treated by multimodality therapy. METHODS AND MATERIALS: Forty two women with FIGO Stages I-IV UPSC who were treated by multimodality therapy were analyzed retrospectively between 1988 and 1998. Data were obtained from tumor registry, hospital, and radiotherapy chart reviews, operative notes, pathology, and chemotherapy flow sheets. All the patients underwent staging laparotomy, peritoneal cytology, total abdominal hysterectomy and salpingo oophorectomy, pelvic and para-aortic lymph node sampling, omentectomy, and cytoreductive surgery, when indicated followed by radiotherapy and/or chemotherapy. Therapy consisted of external beam radiation therapy in 11 patients (26%), systemic chemotherapy in 20 (48%), and both radiotherapy and chemotherapy in 11 (26%). The treatments were not assigned in a randomized fashion. The dose of external beam radiation therapy ranged from 45-50.40 Gy (median 45). Of the 31 patients (74%) who received chemotherapy, 18 received single-agent (58%), whereas 13 received multiagent chemotherapy (42%). RESULTS: Median follow-up for all patients was 19 months (range 4-72). Median follow-up for the surviving patients was 36 months (range 21-72). Their median age was 65 years. Six patients (14%) had Stage I, 8 patients (19%) had Stage II, 10 (24%) had Stage III, and 18 (43%) had Stage IV disease. Twenty-nine patients (69%) had suffered recurrence at the time of last follow-up. The actuarial failure rate at 2 and 5 years was 58% and 67%, respectively. The majority of the patients (19/29) recurred in the abdomen, vagina, or pelvis (66%). Metastases outside the abdomen were much less common as the first site of failure (17%). Twenty-five patients (60%) had died at the time of reporting; the observed survival rate at 2 years and 5 years was 52% and 43%, respectively. CONCLUSIONS: Our data suggest that, after multimodality therapy of FIGO Stage I-IV UPSC, most patients developed abdominopelvic (locoregional) failure, and the great majority of the failures occurred in the abdomen, vagina, and pelvis (66%). Abdominopelvic failure as a component of distant failure occurred in an additional 5 patients (17%). Distant failure alone occurred in 17% of the patients.We propose that future studies should combine whole abdominal radiotherapy (WART) with pelvic and vaginal boosts, in addition to chemotherapy for FIGO Stage I-IV UPSC, especially in patients with minimal residual disease, to attempt to improve the dismal prognosis of patients with UPSC. PMID- 12909236 TI - Pd-ECGF positivity correlates with better survival, while iNOS has no predictive value for cervical carcinomas treated with radiotherapy. AB - PURPOSE: Platelet-derived endothelial cell growth factor (PD-ECGF), which has angiogenic activity, is identical to thymidine phosphorylase. Tumor vascularization is considered to be an important prognostic factor. Nitric oxide synthases (NOSs) are a kind of enzyme that generates nitric oxide. Nitric oxide has not only a self defense against neoplastic cells but also tumor growth stimulation by promoting new blood vessel formation. Our purpose was to investigate the correlation between the expression of PD-ECGF or inducible NOS (iNOS) in cancer cells and prognosis. METHODS AND MATERIALS: Formaldehyde-fixed and paraffin-embedded biopsy specimens excised from 71 cervical squamous cell carcinoma patients who were treated with radiotherapy alone were investigated using an immunohistochemical method. RESULTS: Cancer cells that were positive for PD-ECGF showed intranuclear and cytoplasmic staining patterns. Of the 71 patients, 40 (56%) were positive for PD-ECGF and 31 (44%) were negative. The 5 year survival of the PD-ECGF-positive patients was significantly better than that of the PD-ECGF-negative patients (p = 0.026). Cancer cells that were positive for iNOS showed a cytoplasmic staining pattern. Twenty-seven patients (38%) were positive for iNOS and 44 (62%) were negative. No significant prognostic correlation was observed between iNOS-positive and iNOS-negative patients. CONCLUSION: PD-ECGF positivity in cancer cells is a predictive factor for a good prognosis in cervical squamous cell carcinoma treated with radiotherapy alone. PMID- 12909237 TI - Lymphocyte radiosensitivity correlated with pelvic radiotherapy morbidity. AB - PURPOSE: To test the hypothesis that, before treatment, prostate cancer patients who demonstrate a high yield of ex vivo radiation-induced micronucleus (MN) in G(0) lymphocytes represent a patient population with a greater-than-average risk of developing radiotherapy (RT)-related morbidity. METHODS AND MATERIALS: We prospectively conducted the cytokinesis-block MN assay of peripheral blood lymphocytes (PBLs) in 38 prostate cancer patients. Before the initiation of RT, PBLs from each patient were irradiated (1-4 Gy). The mean patient age +/- SEM was 68.7 +/- 1.0 years. The clinical stage was T1 in 17, T2 in 15, and T3 in 6. The preoperative prostate-specific antigen level was < or =4 ng/mL in 5, 4-10 ng/mL in 18, and >10 ng/mL in 15. All patients underwent standardized pelvic external beam radiotherapy (range 41.4-50.4 Gy) and boost (range 16-26 Gy). The mean follow-up +/- SEM was 32.8 +/- 4.6 months. At the end of follow-up, a radiation oncologist scored the GI or GU morbidity according to the Radiation Therapy Oncology Group criteria without knowledge of the MN data. RESULTS: We found that between the average reactors (n = 25; i.e., patients who had Grade 1 or less RT related morbidity) and over reactors (n = 13; i.e., patients who developed Grade 2 or greater RT-related morbidity), the differences in the ex vivo radiation dose response relationship of MN yield in PBLs were highly significant, especially at doses of > or =2 Gy. Also, the development of RT-related morbidity correlated with the radiation dose-response relationship of MN yield in PBLs before treatment, but did not correlate with any of the patients' clinical variables. CONCLUSION: Our findings suggest that the pre-RT ex vivo radiation dose-response relationship of MN yield in PBLs may be a significant predictive factor for the development of GI or GU morbidity in prostate cancer patients after pelvic RT. PMID- 12909238 TI - Does prone positioning reduce small bowel dose in pelvic radiation with intensity modulated radiotherapy for gynecologic cancer? AB - PURPOSE: Intensity-modulated radiotherapy (IMRT) has been shown to reduce the radiation dose to small bowel in pelvic RT in gynecology patients. Prone positioning has also been used to decrease small bowel dose by displacement of small bowel from the RT field in these patients. The purpose of this study was to determine whether the combination of both IMRT and prone positioning on a belly board can reduce small bowel dose further in gynecologic cancer patients undergoing pelvic RT. METHODS AND MATERIALS: IMRT plans for pelvic RT were computed in 16 patients with gynecologic cancer who had undergone planning CT scans in both the supine and the prone positions on a belly board. For the gross tumor volume, the uterus, cervix, and tumor (or postoperative region) were traced. The clinical target volume was defined as the vessels and lymph nodes from the obturator level to the aortic bifurcation, presacral region, and upper 4 cm of the vagina, in addition to gross tumor volume. The planning target volume was defined as a 2-cm margin in addition to the gross tumor volume and upper 4 cm of the vagina, and 1.5 cm for lymph nodes and vessels. Normal tissue regions of interest included small bowel, large bowel, and bladder. IMRT plans using (1) the limited arc technique (180 degrees arc length) and (2) the extended arc technique (340 degrees arc length) were computed. Dose-volume histograms for normal tissue structures and target were compared between the supine and prone IMRT plans using the paired t test. RESULTS: Prone positioning on a belly board decreased the small bowel dose in gynecologic pelvic IMRT, and the magnitude of improvement depended on the specific IMRT technique used. With the limited arc technique, prone positioning significantly decreased the irradiated small bowel volume at the 25-50-Gy dose levels compared with supine positioning. Small bowel volumes receiving > or =45 Gy decreased from 19% to 12.5% (p = 0.005) with prone positioning. With the extended arc technique, the decrease in irradiated small bowel volume was less marked, but remained detectable in the 35-45-Gy dose levels. Small bowel volumes receiving > or =45 Gy decreased from 13.6% to 10.1% (p = 0.03) with prone positioning. The effect of prone positioning on large bowel and bladder was variable. Large bowel volumes receiving > or =45 Gy increased with prone positioning from 16.5% to 20.6% (p = 0.02) in the limited arc technique and was unaffected in the extended arc technique. CONCLUSION: These preliminary data suggest that prone positioning on a belly board can reduce the small bowel dose further in gynecology patients treated with pelvic RT, and that the dose reduction depends on the IMRT technique used. PMID- 12909239 TI - Intraoperative dural irradiation by customized 192iridium and 90yttrium brachytherapy plaques. AB - PURPOSE: After vertebral or paravertebral tumor resection, tumor cells may remain on the dura. Because a tumoricidal dose is difficult to achieve using external beam radiotherapy without exceeding the spinal cord tolerance, we developed intraoperative applicators to deliver additional dose to the dura. METHODS AND MATERIALS: Eight patients with vertebral or paravertebral tumor underwent conformal external beam radiotherapy, tumor resection, and intraoperative radiotherapy to the dura involved by tumor. At surgery, vertebra, soft tissue, and epidural tumor were resected. A radioactive applicator plaque was placed on the dura to deliver 7.5-15 Gy, and then removed. Vertebral reconstruction and stabilization was completed. Chemotherapy was administered for large, high-grade sarcomas. RESULTS: We progressed through three plaque designs, initially (192)Ir, subsequently liquid (90)Y, and finally (90)Y foil in a semicylindrical polycarbonate plaque, in the treatment of 8 patients. The low-energy (90)Y beta emissions provided a more attractive depth dose profile than that achievable with iridium and gave negligible staff radiation exposure. The (90)Y depth dose measured 29% at 2 mm and 9% at 4 mm from the surface of the foil plaque, with acceptable surface dose homogeneity. The average surface dose rate ranged from 18.7 to 47.6 cGy/min for the iridium plaques and 45.2 to 187.5 cGy/min for the (90)Y plaques. The treatments have been without acute or late neurologic complications. The disease of 6 of 8 patients was locally controlled at median potential follow-up of 24 months. CONCLUSIONS: The (90)Y foil applicator is technically elegant, easy to use, and superior to the earlier models. It has been incorporated into a protocol for spinal tumor treatment. PMID- 12909240 TI - The epidermal growth factor receptor mediates radioresistance. AB - PURPOSE: The epidermal growth factor (EGF) receptor is frequently overexpressed in malignant tumors, and its level is correlated with increased cellular resistance to ionizing radiation. However, no precedent studies have investigated whether expression of EGF receptor would by itself confer on cancer cells resistance to radiation. The current study is aimed to address this question. METHODS AND MATERIALS: A full-length human EGF receptor expression vector was transfected into the OCA-I murine ovarian carcinoma cells for stable clones expressing various levels of EGF receptors. Apoptosis and cell clonogenic survival assays were used to evaluate the sensitivity of the resulting cell clones to ionizing radiation. RESULTS: OCA-I cell clones expressing various levels of EGF receptor (OCA-I EGFR) were obtained. These clones showed an EGF receptor level-dependent increase in resistance to ionizing radiation, measured by apoptosis and cell clonogenic survival assays. Compared with the results for parental OCA-I and control vector-transfected OCA-I cells at the 10% cell survival level, the radioresistance was increased by a factor of 1.60 for EGFR-C5 (high level of EGF receptor expression), 1.37 for EGFR-C3 (intermediate level of EGF receptor expression), and 1.28 for EGFR-C1 (low level of EGF receptor expression). Treatment of the OCA-I EGF receptor transfectants with the anti-EGF receptor monoclonal antibody C225 downregulated the levels of EGF receptor, reduced the phosphorylation levels of EGF receptor downstream substrates (such as Akt and MAPK), and reversed the cellular radioresistance. CONCLUSION: Our results demonstrate that overexpression of the EGF receptor conferred cellular resistance to ionizing radiation. The EGF receptor is thus a valid target for potential radiosensitization. PMID- 12909241 TI - Tumor control probability predictions for genetic radiotherapy. AB - PURPOSE: Genetic radiotherapy, the combination of gene therapy and radiation therapy, for cancer treatment is evolving from laboratory studies to clinical trials. Genetic radiotherapy involves the viral infection of cells that change the sensitivity of transduced cells to radiation. Because there is no patient outcome data for genetic radiotherapy, prospective models are needed to determine the expected benefit of this new modality. Such a prospective model has been developed in this work. METHODS AND MATERIALS: An existing tumor control probability (TCP) calculation model developed for external beam radiotherapy was modified for genetic radiotherapy. Specifically, the (1) transduced fraction and (2) enhancement factor of the transduced cells were included in the model. Parametric studies of the effects of these two variables on TCP for head-and-neck cancer were performed. RESULTS: Using reasonable transduction fraction and enhancement factor values (0.8 and 1.4, respectively), the model predicts an increase in the TCP for genetic radiotherapy over radiotherapy alone by up to 15% for the same radiotherapy dose. The theoretical limit of TCP increase was calculated to be near 70%, which may occur with improved techniques that increase the transduced fraction or because of a strong bystander effect. To maintain existing TCP, dose reductions from 5 Gy (reasonable values) to >30 Gy (ideal case) are predicted for genetic radiotherapy over radiotherapy alone. CONCLUSIONS: Our results indicate that genetic radiotherapy has the potential to significantly improve tumor control over radiotherapy alone. PMID- 12909242 TI - Relative roles of ICAM-1 and VCAM-1 in the pathogenesis of experimental radiation induced intestinal inflammation. AB - PURPOSE: Cell adhesion molecules mediate leukocyte recruitment into the irradiated organs; modulation of this process may protect from radiation damage. Our objective was to characterize the requirement for intercellular adhesion molecule 1 (ICAM-1) and vascular cell adhesion molecule 1 (VCAM-1) in intestinal inflammatory response after abdominal irradiation. METHODS AND MATERIALS: Endothelial ICAM-1 and VCAM-1 expression was determined using radiolabeled antibodies in mice 24 h and 14 days after irradiation with 10 Gy, or sham radiation. Leukocyte-endothelial cell interactions in intestinal venules were assessed using intravital microscopy, and the function of ICAM-1 and VCAM-1 in this process by using blocking antibodies and ICAM-1(-/-) mice. RESULTS: The number of adherent leukocytes significantly increased 24 h after irradiation and remained elevated at 14 days. Treatment with anti-ICAM-1 antibodies and ICAM-1 genetic deficiency significantly reduced leukocyte adhesion 24 h after irradiation. At 14 days after irradiation, both wild-type and ICAM-1(-/-) mice had an upregulation of VCAM-1, expression, and VCAM-1 immunoneutralization, but not ICAM-1 immunoneutralization, significantly reduced leukocyte adhesion. In ICAM-1(-/-) mice, regeneration of the intestinal epithelium was enhanced relative to wild-type mice. CONCLUSIONS: ICAM-1 plays a key role in leukocyte recruitment at early time points after abdominal irradiation, whereas VCAM-1 is the main molecular determinant of leukocyte recruitment at late time points. PMID- 12909243 TI - Unexpected changes of rat cervical spinal cord tolerance caused by inhomogeneous dose distributions. AB - PURPOSE: The effects of dose distribution on dose-effect relationships have been evaluated and, from this, iso-effective doses (ED(50)) established. METHODS AND MATERIALS: Wistar rats were irradiated on the cervical spinal cord with single doses of unmodulated protons (150 MeV) to obtain sharp lateral penumbras, using the shoot-through technique, which employs the plateau of the depth-dose profile rather than the Bragg peak. Two types of inhomogeneous dose distributions have been administered: (1) 2 4-mm fields with 8- or 12-mm spacing between the center of the fields (referred to as split-field) were irradiated with variable single doses and (2) cervical spinal cord was irradiated with various combinations of relatively low doses to a large volume (20 mm) combined with high doses to a small volume (4 mm) (referred to as bath and shower). The endpoint for estimating the dose-response relationships was paralysis of the fore or hind limbs. RESULTS: The split-field experiments (2 x 4 mm) showed a shift in the dose-response curves, giving significant higher ED(50) values of 45.4 Gy and 41.6 Gy for 8- and 12-mm spacing, respectively, compared with the ED(50) of 24.9 Gy for the single 8 mm (same total tissue volume irradiated). These values were closer to the ED(50) for a single 4-mm field of 53.7 Gy. The bath and shower experiments showed a large decrease of the ED(50) values from 15-22 Gy when compared with the 4-mm single field, even with a bath dose as low as 4 Gy. There were no histologic changes found in the low dose bath regions of the spinal cord at postmortem. CONCLUSIONS: Not only the integral irradiated volume is a determining factor for the ED(50) of rat cervical spinal cord, but also the shape of the dose distribution is of great importance. The high ED(50) values of a small region or shower (4 mm) decreases significantly when the adjacent tissue is irradiated with a subthreshold dose (bath), even as low as 4 Gy. The significant shift to lower ED(50) values for induction of paralysis of the limbs by adding a low-dose bath was not accompanied by changes in histologic lesions. These observations may have implications for the interpretation of complex treatment plans and normal tissue complication probability in intensity-modulated radiotherapy. PMID- 12909244 TI - Lethal hepatic injury by combined treatment of radiation plus chemotherapy in rats with thioacetamide-induced liver cirrhosis. AB - PURPOSE: To assess lethal hepatic injury by combined treatment of radiation (RT) plus chemotherapy in a rat model with thioacetamide (TAA)-induced liver cirrhosis. METHODS AND MATERIALS: Male Wistar rats were treated with 0.3 g/L TAA in drinking water. The development of liver cirrhosis was histologically confirmed, and the degree of liver function impairment was assessed by indocyanine green retention test (ICG R15). The established cirrhotic rats were given one of the following treatments: partial liver radiotherapy (25 Gy on about one-third of the whole liver), 5-fluorouracil (5-FU) chemotherapy (50 mg/kg), and combined treatment of partial RT plus 5-FU. The treated rats were closely followed until either death or 30 weeks after the treatment, and the postmortem liver sampling was examined for lethal hepatic injury by the treatments. RESULTS: The rats developed overt liver cirrhosis after 30 weeks of TAA treatment. At that time, the mean ICG R15 level in the TAA-treated rats (TAA-rats) was 14.1% +/- 0.7% compared to 4.6% +/- 0.7% in the control (p < 0.05). The 30-week survival rates in the control and TAA-rats were 100% (5/5) and 75% (6/8), respectively, after partial liver RT (p = 0.72). In the 5-FU chemotherapy group, the survival in TAA-rats was only one-half of that in the controls (100% vs. 50%, p = 0.06). Poor survival in TAA-rats was shown also in the combined group of partial RT plus 5-FU (87.5% vs. 16.7%, p = 0.06). The rats that died before the last observation time showed advanced cirrhosis with areas of lobular collapse, in contrast to the moderate cirrhotic features in those that survived. CONCLUSION: In a rat cirrhosis model with mildly impaired liver function, combined treatment of partial RT plus 5-FU resulted in significantly high incidence of lethal liver injury. The results in this study show that a combined treatment of RT plus chemotherapy in cirrhotic patients should be applied with extreme caution. PMID- 12909245 TI - Radiosensitization by pemetrexed of human colon carcinoma cells in different cell cycle phases. AB - PURPOSE: The novel folate antimetabolite Alimta (pemetrexed disodium, LY231514) exhibits antitumor activity in a broad array of human malignancies and was recently found to enhance radiation-induced cell killing in vitro. In the present study, a possible cell cycle phase-specific radiosensitization by pemetrexed was assessed. METHODS AND MATERIALS: Widr human colon carcinoma cells were synchronized by serum withdrawal/stimulation that yielded about 80% cells with G1 DNA content 6 h after replating and more than 60% S-phase cells after 22 h, as assessed by flow cytometry. The respective cultures were irradiated with doses up to 12 Gy in combination with a subtoxic pemetrexed exposure (1.06 microM for 2 h: about 80% survival), or after mock treatment. Survival curves were generated by the clonogenic assay; apoptosis was measured by sub-G1 DNA flow cytometry. RESULTS: The combination treatment of the G1 cells and of the more radioresistant S-phase cell preparations yielded survival rates that were lower than expected for independent cell killing. Radiosensitization, calculated as the ratio of the mean inactivation doses without or with drug exposure (enhancement ratio), was not significantly different for the two cell preparations (enhancement ratio of 2.1 and 2.3, respectively) and was similar to the previously reported value for log-phase cells. Pemetrexed exposure was unable to stimulate an apoptotic response of these cells to radiation. CONCLUSIONS: Radiosensitization by pemetrexed is not cell cycle phase-specific, and the relative radioresistance of S-phase cells is retained. Apoptosis seems to have no influence on radiosensitization in this cell line. PMID- 12909246 TI - Is it necessary to eliminate the posterior dose margin in prostate brachytherapy to achieve an acceptably low risk of late rectal morbidity? AB - PURPOSE: The use of a posterior dose margin in (125)I prostate brachytherapy is controversial. The posterior margin is often eliminated to lower the risk of late rectal morbidity (Radiation Therapy Oncology Group protocols 9805 and P-0019), but this may compromise the posterior prostate dose coverage. The purpose of this work is to determine whether it is necessary to eliminate the posterior margin to achieve an acceptably low risk of Grade 2 (bleeding/ulceration) late rectal morbidity. METHODS AND MATERIALS: The present work is an extension of a previous study in which we reported the probability of Grade 2 late rectal morbidity (bleeding/ulceration) as a function of the maximum rectal dose. Here, we define the relationship between the maximum rectal dose and the width of the posterior dose margin. From this relationship, and the probability of late morbidity determined earlier, we assessed the probability of late rectal morbidity as a function of the width of the posterior dose margin. The present work was based on the preplans of 15 permanent (125)I prostate seed implants having volumes ranging from 19 to 78 cm(3). All used peripheral loading with (125)I sources in the range 0.35-0.45 mCi. The maximum rectal dose was taken to be the isodose line coincident with the posterior edge of the prostate. The maximum rectal dose and the corresponding margin width were determined from the isodose distribution. The sensitivity of the relationship between the maximum rectal dose and the margin width to the prostate volume, the seed density, and the distance of the most posterior seeds from the posterior edge of the prostate was investigated. RESULTS: The maximum rectal dose is directly proportional to the margin width. This relationship is relatively insensitive to the prostate volume and the seed density, but is sensitive to the location of the posterior seeds relative to the posterior edge of the gland. Moving the seeds from 5 mm to 3 mm from the edge typically increased the maximum rectal dose by 17%. With the posterior seeds 3 mm from the edge, the maximum rectal doses that corresponded to 1 mm, 2 mm, 3 mm, 4 mm, and 5 mm margins were 187 +/- 6 Gy, 222 +/- 8 Gy, 257 +/- 11 Gy, 292 +/- 14 Gy, and 327 +/- 17 Gy, respectively. The corresponding probabilities that a patient will experience late rectal morbidity are < or =1%, < or =2%, < or =3%, < or =5%, and < or =7%, respectively. CONCLUSIONS: Our results indicate that a 2-3 mm posterior dose margin can be used in prostate brachytherapy with a relatively low (2-3%) risk of Grade 2 (bleeding/ulceration) late rectal morbidity, provided the sources in the posterior row are implanted at least 3 mm from the edge of the prostate. A practical guideline is to keep the maximum rectal dose below 150% of the target dose. PMID- 12909247 TI - In regard to Solomon et al.: EGFR blockade with ZD1839 ("Iressa") potentiates the antitumor effects of single and multiple fractions of ionizing radiation in human A431 squamous cell carcinoma. IJROBP 2003;55:713-723. PMID- 12909250 TI - A two-current model for the dynamics of cardiac membrane. AB - In this paper we introduce and study a model for electrical activity of cardiac membrane which incorporates only an inward and an outward current. This model is useful for three reasons: (1) Its simplicity, comparable to the FitzHugh-Nagumo model, makes it useful in numerical simulations, especially in two or three spatial dimensions where numerical efficiency is so important. (2) It can be understood analytically without recourse to numerical simulations. This allows us to determine rather completely how the parameters in the model affect its behavior which in turn provides insight into the effects of the many parameters in more realistic models. (3) It naturally gives rise to a one-dimensional map which specifies the action potential duration as a function of the previous diastolic interval. For certain parameter values, this map exhibits a new phenomenon--subcritical alternans--that does not occur for the commonly used exponential map. PMID- 12909251 TI - Optimality of collective choices: a stochastic approach. AB - Amplifying communication is a characteristic of group-living animals. This study is concerned with food recruitment by chemical means, known to be associated with foraging in most ant colonies but also with defence or nest moving. A stochastic approach of collective choices made by ants faced with different sources is developed to account for the fluctuations inherent to the recruitment process. It has been established that ants are able to optimize their foraging by selecting the most rewarding source. Our results not only confirm that selection is the result of a trail modulation according to food quality but also show the existence of an optimal quantity of laid pheromone for which the selection of a source is at the maximum, whatever the difference between the two sources might be. In terms of colony size, large colonies more easily focus their activity on one source. Moreover, the selection of the rich source is more efficient if many individuals lay small quantities of pheromone, instead of a small group of individuals laying a higher trail amount. These properties due to the stochasticity of the recruitment process can be extended to other social phenomena in which competition between different sources of information occurs. PMID- 12909252 TI - Life history implications of allocation to growth versus reproduction in dynamic energy budgets. AB - We compare the implications of determinate vs. indeterminate growth of a parthenogenetic iteroparous ectotherm at constant food density in the context of the dynamic energy budget theory, which specifies the tight links between life history traits, such as feeding, aging, growth and reproduction. We do a comparative analysis using, as measure of fitness, the life span reproduction, the population growth rate, and the conversion efficiency of food to biomass. When extrinsic mortality is constant, indeterminate growth cannot maximize fitness if measured by the population growth rate or the conversion efficiency, except when mortality is low, in which case both types of animals are similar. If the fitness measure is life span reproduction, indeterminate growth maximizes fitness even with constant mortality, provided it is not very high. When mortality decreases with size, indeterminate growth maximizes fitness for almost all measures of fitness. Finally, we suggest an evolutionary link between allocation strategies and expected life span. In populations of long living species, each type of animal can establish in the population of the other. In populations of short living species, determinate growers can invade, and displace, a population of indeterminate ones. However, when the mortality risk of organisms with small size is much higher than those of large size, indeterminate growers can be superior. PMID- 12909253 TI - An illustration of variable precision rough set theory: the gender classification of the European barn swallow (Hirundo rustica). AB - This paper introduces a new technique in the investigation of object classification and illustrates the potential use of this technique for the analysis of a range of biological data, using avian morphometric data as an example. The nascent variable precision rough sets (VPRS) model is introduced and compared with the decision tree method ID3 (through a 'leave n out' approach), using the same dataset of morphometric measures of European barn swallows (Hirundo rustica) and assessing the accuracy of gender classification based on these measures. The results demonstrate that the VPRS model, allied with the use of a modern method of discretization of data, is comparable with the more traditional non-parametric ID3 decision tree method. We show that, particularly in small samples, the VPRS model can improve classification and to a lesser extent prediction aspects over ID3. Furthermore, through the 'leave n out' approach, some indication can be produced of the relative importance of the different morphometric measures used in this problem. In this case we suggest that VPRS has advantages over ID3, as it intelligently uses more of the morphometric data available for the data classification, whilst placing less emphasis on variables with low reliability. In biological terms, the results suggest that the gender of swallows can be determined with reasonable accuracy from morphometric data and highlight the most important variables in this process. We suggest that both analysis techniques are potentially useful for the analysis of a range of different types of biological datasets, and that VPRS in particular has potential for application to a range of biological circumstances. PMID- 12909254 TI - A region-based model framework for the rat urine concentrating mechanism. AB - The highly structured organization of tubules and blood vessels in the outer medulla of the mammalian kidney is believed to result in preferential interactions among tubules and vessels; such interactions may promote solute cycling and enhance urine concentrating capability. In this study, we formulate a new model framework for the urine concentrating mechanism in the outer medulla of the rat kidney. The model simulates preferential interactions among tubules and vessels by representing two concentric regions and by specifying the fractions of tubules and vessels assigned to each of the regions. The model equations are based on standard expressions for transmural transport and on solute and water conservation. Model equations, which are derived in dynamic form, are solved to obtain steady-state solutions by means of a stable and efficient numerical method, based on the semi-Lagrangian semi-implicit method and on Newton's method. In this application, the computational cost scales as O(N2), where N is the number of spatial subintervals along the medulla. We present representative solutions and show that the method generates approximations that are second-order accurate in space and that exhibit mass conservation. PMID- 12909255 TI - Regression and regrowth of tumour cords following single-dose anticancer treatment. AB - In this paper, the evolution of a tumour cord after treatment is investigated by extensive numerical simulations on the basis of a mathematical model developed by Bertuzzi et al. (submitted). The model is formulated in cylindrical symmetry adopting the continuum approach, and takes into account the influence of oxygen level on the proliferation and death rate of cells, the volume reduction due to disgregation of dead cells, and the cell killing effects of radiation and drugs. Some extensions of the model are proposed to represent more accurately the radioresistance of hypoxic cells and the cytotoxic action of anticancer drugs. The steady state of the cord, and the cord evolution from the steady state after the delivery of a single dose of an anticancer agent, are computed for various combinations of model parameters and for different choices of the functions describing the effects of treatments. The results of the numerical computations show that, in spite of its many simplifications, the model behaviour appears to be reasonable in view of the available experimental observations. The model allows having a better insight into some complex treatment-related events, such as cell reoxygenation and repopulation. PMID- 12909256 TI - Modelling of signalling via G-protein coupled receptors: pathway-dependent agonist potency and efficacy. AB - A mathematical model is constructed to study promiscuous coupling of receptors to G-proteins and to simulate events leading to the activation of multiple effector pathways within a cell. The model is directed at a better understanding of the factors that determine the efficacy and potency of a drug. Assuming that the receptors can exist in multiple conformational states, and allowing for agonist specific conformation, a system of ordinary differential equations is constructed and subsequently pathway-dependent agonist efficacy and potency order is predicted. A simple case of the compartmentalization of receptors and G-proteins is also given, using the current model to illustrate the effects of spatial heterogeneity on the predicted response. PMID- 12909259 TI - The use of consumption data to assess exposure to biotechnology-derived foods and the feasibility of identifying effects on human health through post-market monitoring. AB - The pre-market safety assessment of foods derived through biotechnology provides a scientific basis for concluding reasonable certainty of no harm and ensuring safety. At a minimum, the outcome of such an assessment provides sufficient information to estimate the likelihood of adverse effects on consumers, generally precluding the need for post-market monitoring. Post-market monitoring (PMM) may be appropriate under certain conditions where a better estimate of dietary exposure and/or nutritional consequence of a biotechnology-derived food is required, when a potential safety issue, such as allergenicity, cannot be adequately addressed through pre-market studies, or to corroborate dietary intakes of a nutritionally improved food with beneficial effects on human health. Monitoring programs must be hypothesis-driven, and are dependent upon the availability of accurate consumption data. Exposure assessment methods include both deterministic and probabilistic estimates of intakes using food supply data, individual dietary surveys, household surveys, or total diet studies. In the development of a monitoring approach, resource allocation should be dependent upon both the desired level of conservatism and the endpoint of interest. However, the cost of monitoring varies substantially, and the potential to determine causation may be limited. PMID- 12909260 TI - Assessing the zearalenone-binding activity of adsorbent materials during passage through a dynamic in vitro gastrointestinal model. AB - A novel approach is presented herein to study the intestinal absorption of mycotoxins by using a laboratory model that mimics the metabolic processes of the gastrointestinal (GI) tract of healthy pigs. This model was used to evaluate the small-intestinal absorption of zearalenone from contaminated wheat (4.1 mg/kg) and the effectiveness of activated carbon and cholestyramine at four inclusion levels (0.25, 0.5, 1 and 2%) in reducing toxin absorption. Approximately 32% of ZEA intake (247 microg) was released from the food matrix during 6 h of digestion and was rapidly absorbed at intestinal level. A significant reduction of intestinal absorption of ZEA was found after inclusion of activated carbon or cholestyramine, even at the lowest dose of adsorbents, with a more pronounced effect exhibited by activated carbon. In particular, when 2% of activated carbon or cholestyramine was added to the meal the ZEA intestinal absorption was lowered from 32% of ZEA intake to 5 and 16%, respectively. The sequestering effect of both adsorbents took place already during the first 2 h of digestion and persisted during the rest of the experiment. The GI-model is a rapid and physiologically relevant method to test the efficacy of adsorbent materials in binding mycotoxins and can be used to pre-screen mycotoxin/adsorbent combinations as an alternative to animal experiments. PMID- 12909261 TI - Reduction of genistein clastogenicity in Chinese hamster V79 cells by daidzein and other flavonoids. AB - A broad spectrum of health benefits has been ascribed to soy products. These products contain soy protein and relatively high levels of polyphenolic compounds known as flavonoids. While they are the most likely candidates for biological activity, flavonoids as a class, and of specific interest, genistein, are well known to be genotoxic due to their ability to "poison" cellular DNA topoisomerase II (topo II) resulting in stable chromosome breakage and mutation and raising questions about the long term health effects associated with chronic flavonoid exposure. Interestingly, some flavonoids, such as biochanin, galangin and daidzein, are catalytic topo II inhibitors (not poisons) and actually antagonize the clastogenicity of topo II poisons. It is shown in the present paper that flavonoids possessing catalytic topo II inhibitory activity, strongly antagonize the clastogenicity of genistein in Chinese hamster V79 cells. Importantly, one of these, daidzein, is a major constituent of marketed soy products. It is conjectured that the potential human clastogenic risk of soy products containing genistein might be mitigated or abolished due to the presence of daidzein or other flavonoids in those products. PMID- 12909262 TI - Lack of clastogenic effects in cultured human lymphocytes treated with hydroquinone. AB - Hydroquinone (HQ) occurs in the environment as a result of manmade processes as well as in natural products from plants and animals. The compound has been reported to produce chromosomal effects in some in vivo and in vitro animal models. However, its potential to produce similar effects in human lymphocytes is less clear. To obtain more information on the clastogenic potential of HQ in human cells, its ability to induce structural chromosomal aberrations in human lymphocytes in vitro has been examined, both in the absence and presence of exogenous metabolic activation. Moreover, the effect of HQ pre-incubation on peroxide induced clastogenicity was studied, because HQ has putative chemopreventive activity as well. It was found that HQ was cytotoxic, but did not induce chromosomal aberrations in human lymphocytes cultured in vitro. Additionally, it was observed that pre-incubation of lymphocytes with HQ resulted in a concentration dependent reduction of the H2O2 induced chromosomal aberrations (P=0.069). However, this effect was present at 12 mM H2O2 only, because of high cytotoxicity at higher dosages. PMID- 12909263 TI - A rapid and sensitive cytotoxicity screening assay for trichothecenes in cereal samples. AB - The application of a bioassay for toxicity screening of cereal samples has been studied. The sensitive Swiss mouse 3T3 fibroblasts (3T3 cells) were used to determine the cytotoxicity of Fusarium trichothecenes T-2 toxin (T-2), HT-2 toxin (HT-2), deoxynivalenol (DON) and nivalenol (NIV) in cereal samples. Clean-up of the sample extracts was performed using the MycoSep #225 column and the cytotoxic effects of the samples were determined using the 5-bromo-2'-deoxyuridine (BrdU) bioassay assessing DNA-synthesis. Trichothecene-free samples of wheat, wheat bran and barley had no toxic effect on the 3T3 cells at sample concentrations up to 400 mg/ml cell culture medium while a few oat samples had an inhibiting effect on the DNA-synthesis. IC(50) values (50% response compared with untreated cells) of trichothecene-free wheat and oats spiked with T-2, HT-2, DON and NIV were similar to IC(50) values of pure toxins. A cytotoxicity screening of 33 different cereal samples showed that all samples contaminated with a trichothecene concentration higher than the IC(50) values of T-2, HT-2, DON and NIV had a clear inhibiting effect on DNA-synthesis of the 3T3 cells. The culture of 3T3 cells on microtiter plates in combination with the simple and rapid sample clean-up using the MycoSep #225 column provides a sensitive and easy method for the screening of trichothecenes in cereal samples. PMID- 12909265 TI - Cytogenetic toxicity of neem. AB - The cytogenetic toxicity of the leaf extract of neem was evaluated in murine germ cells. The extract was found to induce structural and numerical changes in the spermatocyte chromosomes as well as synaptic disturbances in them at their first metaphase. A significant increase in the frequency of sperms with abnormal head morphology and the decrease in mean sperm count were also observed. This spermatotoxic effect of the neem extract corroborates its germ cell mutagenicity. The possible role of azadirachtin, the most active principle present in the neem extract, in producing the observed genotoxic effect is discussed. PMID- 12909264 TI - Effects of dietary antioxidants and 2-amino-3-methylimidazo[4,5-f]- quinoline (IQ) on preneoplastic lesions and on oxidative damage, hormonal status, and detoxification capacity in the rat. AB - The potential beneficial or adverse affect of prolonged dietary administration of moderate to high doses (1-100 mg/kg diet) of the antioxidants, lycopene, quercetin and resveratrol or a mixture of lycopene and quercetin was investigated in male F344 rats. Selected markers for toxicity and defense mechanisms were assayed in blood, liver and colon and the impact of the antioxidant administrations on putative preneoplastic changes in liver and colon was assessed. The dietary carcinogen, 2-amino-3-methylimidazo[4,5-f]quinoline (IQ) (200 mg/kg diet) served as a pro-oxidant, genotoxicity and general toxicity control. IQ increased the levels of protein and DNA oxidation products in plasma, the area of glutathione S-transferase-placental form positive (GST-P) foci in the liver as well as the number of colonic aberrant crypt foci (ACF). All antioxidants and the antioxidant combination significantly increased the level of lymphocytic DNA damage, to an extent comparable with the effect induced by IQ. In contrast to the control group where no GST-P foci were detected, GST-P foci were detected in animals exposed to quercetin, lycopene and the combination of the two. However, the increase in the volume of GST-P foci did not reach statistical significance. The present results indicate that moderate to high doses of common dietary antioxidants can damage lymphocyte DNA and induce low levels of preneoplastic liver lesions in experimental animals. Long-term exposure to moderate to high doses of antioxidants may thus via pro-oxidative mechanisms and non-oxidative mechanisms modulate carcinogenesis. PMID- 12909266 TI - Irradiation effects on embryotoxicity and oxidative properties of gossypol dissolved in methanol. AB - Gossypol dissolved in methanol (0.25 and 0.5 mg/ml) was gamma-irradiated at 0, 5, 10, and 20 kGy. The gossypol content was significantly reduced by irradiation in a dose-dependent manner. Ames test performed with nonirradiated and irradiated gossypol solutions was negative at the level of 0.1, 0.5, 1, 10, 50, and 100 microg gossypol/plate. At the gossypol concentration of 2.5 microg/ml, the blastocysts formation rate of mice embryo were decreased by a factor of two as compared with the control, but irradiation at 20 kGy increased the blastocysts formation, resulting in no difference from the control. Addition of gossypol to oil emulsion system or oil showed a prooxidative effect of lipids at the beginning stage by increasing 2-thiobarbituric acid reactive substances (TBARS) and peroxide value but irradiation of the gossypol solution decreased the oxidative changes significantly as compared with the nonirradiated one. Results indicated that irradiation decreased the gossypol concentration dissolved in methanol, resulting in a reduction of embryotoxicity in mice. PMID- 12909267 TI - A 13-week subchronic toxicity study of paprika color in F344 rats. AB - The fruit of the paprika (Capsicum annuum) has been widely used in various countries as a spice and food-coloring additive. As a part of the safety assessment of paprika color (Paprika oleoresin), a 13-week subchronic toxicity study was performed in F344 rats. To establish a no-observed-adverse-effect level (NOAEL) for application in subsequent long-term studies, rats were fed powder diet containing paprika color at dose levels of 0 (basal diet), 0.62, 1.25, 2.5 and 5% (maximum) for 13 weeks. During the experiment, there were no remarkable changes in general appearance and no deaths occurred in any experimental group. Although serum total cholesterol was dose-dependently increased in both sexes, no related histopathological changes were observed in the liver. Slight inflammatory cell infiltration in the myocardium and vacuolation of hepatocytes were noted in both control and paprika color-treated animals, but there were no clear differences between groups. In conclusion, paprika color even at 5% in the diet (0.67 g/rat/day or 2948.4 mg/kg bw/day for male rats and 0.43 g/rat/day or 3197.4 mg/kg bw/day for female rats) did not cause remarkable adverse effects in F344 rats. Thus, the NOAEL and the maximum dose level for carcinogenicity testing of paprika color were concluded to be 5% in the diet. PMID- 12909268 TI - Toxicological effects of chronic low doses of aflatoxin B(1) and fumonisin B(1) containing Fusarium moniliforme culture material in weaned piglets. AB - The effects of chronic oral exposure (28 days) to aflatoxin B(1) (AFB(1)) and fumonisin B(1) (FB(1)) were studied in weaned piglets. Six experimental groups, each comprising two neutered males and two females, were fed ad libitum with rations containing: (A) 0 mg of FB(1) and 0 mg of AFB(1)/kg of feed (control); (B) 10 mg of FB(1)/kg of feed; (C) 30 mg of FB(1)/kg of feed; (D) 50 microg of AFB(1)/kg of feed; (E) 10 mg of FB(1) plus 50 microg of AFB(1)/kg of feed; (F) 30 mg of FB(1) plus 50 microg of AFB(1)/kg of feed. The animals were inspected twice daily and their body weight and feed consumption were recorded weekly and daily, respectively. Samples of feces and urine were collected 24 h after the start of the experiment, to check for fumonisin residues by HPLC analysis. Blood samples were drawn at the start of the experiment and after 28 days for quantification of hematological and biochemical parameters. Necropsies were performed after 28 days; at necropsy, the organs were weighed, inspected macroscopically and processed for histopathological and toxicological analyses. All piglets from groups C and F presented typical signs of pulmonary edema, with reduced feed consumption and body weight gain as well as pathological alterations. FB(1) was detected in feces and urine at 24 h of intoxication and in liver after 28 days of intoxication. Increases were detected regarding the following hematological and biochemical parameters in animals from treatments C and F: erythrocyte number; hematocrit; total bilirubin; total protein; activity of serum alkaline phosphatase, aspartate aminotransferase, and alanine aminotransferase. Cholesterol levels were significantly aumented only in animals from groups C and F, whereas albumin concentrations increased in groups C, F, B and E. The average organ/body weight ratio of piglets (hearth, liver and lung) were significantly greater in groups C and F. The only joint effects of FB(1) and AFB(1) detected (group F) were a decrease in feed consumption during the last week of intoxication and in feed conversion throughout the 28 days of intoxication. Chronic intoxication of piglets with AFB(1) and FB(1) leads to important losses of productivity. PMID- 12909269 TI - An investigation of the general, reproductive and postnatal developmental toxicity of Betapol, a human milk fat equivalent. AB - Betapol consists of triglyceride fatty acids commonly found in vegetable and animal fats. A similarity to human milk fat indicated a potential use in infant formulae as well as for food use in general. To test the potential for substantial equivalence with a related food grade oil, palm oil, Betapol was fed to rats at 15% content in the diet using an augmented two-generation study, in order to obtain information on general (6 months), reproductive and postnatal developmental toxicity in a single study rather than separate studies. For comparison there were two control groups, namely a comparative control fed a diet containing 15% of food grade oil and a negative, or laboratory standard control fed a commercial rodent breeding diet (LAD), containing 2.3-4.7% fat. It was reasoned that if Betapol fed groups showed differences from the comparative control in the direction of the negative control these would reflect differences in the nutritional value of the high fat diets. Presence of a toxicant might be indicated by differences from the comparative control group opposite to the negative control group. The group fed 15% Betapol showed occasional, statistically significant differences from the comparative control group but the direction of difference was towards the negative control group and did not indicate the presence of an unexpected toxicant. PMID- 12909270 TI - Characterization of coloured compounds obtained by enzymatic extraction of bakery products. AB - Melanoidins, the brown-colored polymers formed through Maillard type reaction in several heat-treated foods, represent a significant part of our diet, with an average intake of grams per day. Most of the studies on the physiological effects of these compounds have been performed using the water soluble melanoidin fractions. But dietary melanoidins formed on the surface of bakery products are poorly soluble in water as well as in organic solvents. In this work, an enzymatic solubilization procedure was developed on a gluten-glucose model system and it was applied to bread and biscuits. The soluble material obtained was tested for its antioxidant activity, for its effect on phase-I and phase-II xenobiotic enzymes and for potential cytotoxic effects. Soluble melanoidins from model system and biscuits exhibit a strong antioxidant activity and do not show any cytotoxicity on Caco-2 cells. Melanoidins extracted from biscuits was able to inhibit the activity of Phase I (NADPH-cytochrome-c reductase) and Phase II (Glutathione-S-transferase) enzymes, whereas the low molecular weight melanoidins isolated from gluten-glucose model system inhibit the activity of NADPH cytochrome-c reductase. PMID- 12909271 TI - Dose-dependent effects of glutamate in pyridoxine-induced neuropathy. AB - In order to explore the effects of glutamate in a pyridoxine megadose-induced neuropathy, rats were received glutamate either 0.5 or 1 g/kg/day orally with or without pyridoxine 0.8 g/kg/day intraperitoneally for 14 days. The animal's motor coordination, the muscle power and the thermal threshold were observed daily. The nerve conduction velocity was measured at day 0 and day 15 of the treatment. Glutamate either 0.5 or 1 g/kg/day appeared to have no effect on motor coordination, the nerve conduction velocity and the muscle power score compared with control. However, the thermal response latency was significantly decreased (from day 9) in animals treated with 1 g/kg/day glutamate. In pyridoxine-induced neuropathy rats, glutamate 0.5 g/kg/day significantly decreased the effects of pyridoxine on the sciatic nerve conduction velocity, the muscle power score and the motor coordination. Interestingly, glutamate at a dose of 1 g/kg/day worsened the neurotoxic effects cause by pyridoxine. PMID- 12909272 TI - Modulation of lipopolysaccharide-induced proinflammatory cytokine production in vitro and in vivo by the herbal constituents apigenin (chamomile), ginsenoside Rb(1) (ginseng) and parthenolide (feverfew). AB - Dietary supplements are not subject to the same pre-market approval as conventional drugs, thus the true efficacy and, in cases, safety of these products is not known. The objective of this study was to evaluate the potential anti-inflammatory properties of three herbal constituents, apigenin (chamomile), ginsenoside Rb(1) (ginseng) and parthenolide (feverfew) on lipopolysaccaharide (LPS)-induced proinflammatory cytokine production, and to determine if effects in cell culture could predict results in an intact animal model. Murine macrophage cells and mice were treated with the stimulant LPS and herbal constituents, and resultant culture supernatant and serum, respectively, were evaluated for interleukin (IL)-6 and tumor necrosis factor (TNF)-alpha by ELISA. All three constituents inhibited LPS-induced IL-6 and/or TNF-alpha production in culture. Inhibition of these two cytokines was observed in mice, but did not display the same patterns of inhibition as cell culture data. The results suggest that all three constituents possessed anti-inflammatory properties, but that cell culture data can only be used to approximate potential effects in animals, and must be confirmed using appropriate animal models. PMID- 12909273 TI - Aorta wall quantitative alterations due to different long-term high-fat diet in rats. AB - Twenty Wistar male rats, with ages varying from 21 days to 12 months, were fed with diets supplemented by soybean oil (S group), canola oil (CA group), lard and egg yolk (LE group), and canola plus lard and egg yolk (CA+LE group). The LE group presented the highest heart mass/body mass ratio and the highest blood pressure as well. The aortic structure suffered a number of alterations, from CA rats group (minor alterations) to LE and CA+LE rats group (major alterations). The CA group showed the smallest aorta thickness. No significant differences among groups were found concerning the number of aortic lamellae and the density of tunica media smooth muscle nuclei. Surface density of the aortic lamellae was greater in the CA group and smaller in the LE group. These results suggest that the dietary intake of canola oil could attenuate the aorta structure aging process with a favorable BP maintenance. PMID- 12909275 TI - Toxicity of Australian essential oil Backhousia citriodora (lemon myrtle). Part 2. Absorption and histopathology following application to human skin. AB - The in vitro percutaneous absorption of the essential oil of lemon myrtle (Backhousia citriodora) has been studied in freshly excised human full-thickness abdominal skin obtained from patients undergoing elective surgery. Absorption of lemon myrtle oil in human skin discs (4.9 cm(2)) was evaluated using a Franz cell diffusion system following topical application of neat lemon myrtle oil (100 microl or 18.29 mg/cm(2)) to the epidermal surface at exposure durations of 1 to 12 h. Gas chromatography mass spectrometry (GCMS) was used as an analytical technique to determine the absorption of lemon myrtle oil components in full thickness skin. Citral; (consisting of isomers, geranial and neral) was the only component of lemon myrtle oil found to be absorbing into skin at all exposure periods. At the maximum exposure duration of 12 h, the total absorption of citral in the full-thickness skin disc was 0.29+/-0.07 mg/cm(2) (mean+/-S.D., n=4) of the applied dose. Although the absorption of lemon myrtle oil components was limited, haematoxylin and eosin (H&E) staining showed significant losses in the cellular functioning of skin including; losses of integrity and solubilisation of the stratum corneum, cellular necrosis (to 15%) and cellular vacuolation (to 25%) on comparison to control skin. When a formulated product containing 1% lemon myrtle oil (0.18 mg/cm(2)) was applied to human skin discs (4.9 cm(2)) at 8 h exposure the total absorption of citral in the full-thickness skin disc was 5.12+/-0.60 x 10(-4) mg/cm(2) (mean+/-S.D., n=4) of the applied dose. No other components were detected. The histopathological assessment indicated limited damage to epidermal cells. The combination of the above methodologies enabled the generation of data that could be used for a comprehensive evaluation of the toxicity effects of lemon myrtle oil for topical application. PMID- 12909274 TI - Lycopene oxidation product enhances gap junctional communication. AB - Carotenoids as well as their metabolites and oxidation products stimulate gap junctional communication (GJC) between cells, which is thought to be one of the protective mechanisms related to cancer-preventive activities of these compounds. Increased intake of lycopene by consumption of tomatoes or tomato products has been epidemiologically associated with a diminished risk of prostate cancer. Here, we report a stimulatory effect of a lycopene oxidation product on GJC in rat liver epithelial WB-F344 cells. The active compound was obtained by complete in vitro oxidation of lycopene with hydrogen peroxide/osmium tetroxide. For structural analysis high performance liquid chromatography, gas chromatography coupled with mass spectrometry, ultraviolet/visible-, and infrared spectrophotometry were applied. The biologically active oxidation product was identified as 2,7,11-trimethyl-tetradecahexaene-1,14-dial. The present data indicate a potential role of lycopene degradation products in cell signaling enhancing cell-to-cell communication via gap junctions. PMID- 12909276 TI - Environmental boron contamination in waters of Hisarcik area in the Kutahya Province of Turkey. AB - Our aim was to determine boron levels in waters of rich boron mine areas in Turkey. There is a naturally occurring high level of boron in the ground waters of some parts of Turkey. Our study area is Hisarcik village in the Kutahya Province of Turkey which has a large number of boron mines. Boron occurs in combined form, usually borax (Na2B4O7 x 10H2O) and colemanite (Ca2B6O11 x 5H2O). Water samples were collected from many springs in order to determine boron concentrations and the amount of boron that is exposed to 88 samples from the drinking water (tap water), 37 from ground water and 257 from stream or pond water (surface water). Urine samples taken from 42 residents in this area were analyzed for boron and compared with its levels in drinking waters. The inductively coupled plasma/mass spectrometry (ICP/MS) method was used for the analysis of boron concentrations. Water boron contents at differing locations of this area ranged from 2.05 to 29.00 mg B/l. Mean value of boron was 10.20+/-4.08 mg B/l. Total urinary boron excretion was ranged from 0.04 to 50.70 mg B/l and mean value was 8.30+/-10.91 mg B/l. Current information on sources, occurrence, pollution potential and toxicity of environmental boron is discussed. PMID- 12909277 TI - Ripe fruit of Solanum nigrum L. inhibits cell growth and induces apoptosis in MCF 7 cells. AB - Solanum nigrum L. (SNL) has been traditionally used as a herbal plant, whose fruit is believed to have anti-tumor properties, although the mechanism for the activity remains to be elucidated. In this study, we prepared an ethanol extract from ripe fruits of SNL and investigated the mechanism involved in its growth inhibitory effect on MCF-7 human breast cancer cells. Results from proliferation assay using tritium uptake showed that the proliferative capacity of MCF-7 cells was strongly suppressed in the presence of SNL ethanol extract. This was further confirmed through MTT assay and trypan blue exclusion experiments, which showed a very close correlation between the SNL extract concentration and the surviving cell numbers. The SNL extract-mediated suppression of cell growth was verified to be apoptotic, based on the appearance of DNA laddering, increase in DNA fragmentation, and low fluorescence intensity in nuclei after propidium iodide staining of the cells. Furthermore, the SNL extract was revealed to be a potential scavenger of hydroxyl radicals and DPPH radicals rather than superoxide anions. Collectively, our findings suggest that SNL fruit extract could be used as an anti-oxidant and cancer chemo-preventive material. PMID- 12909278 TI - Effects of bis(2-ethylhexyl) phthalate (DEHP) on secondary sex ratio of mice in a cross-mating study. AB - Bis(2-ethylhexyl) phthalate (DEHP) was given in the diet to provide levels of 0 (C) or 0.03% (T) from 5 weeks of age of the F(0) generation to birth of the F(1) generation in mice. At 9 weeks of age, each female was paired with one male from the same or another treatment groups (cross-mating: C/C, T/C, C/T, T/T), for a period of 5 days. The males were removed from females after 5 days, and the females were allowed to carry their litters to term and deliver. There were no adverse effects of DEHP on either litter size, litter weight and sex ratio at birth. The average body weight of male offspring was significantly increased in all treatment groups at birth. There were no adverse effects of DEHP on female offspring weight at birth. The dose level of DEHP in the present study produced no adverse effects on secondary sex ratio, which meant sex ratio at birth, in mice. PMID- 12909279 TI - Neurodegeneration in amyotrophic lateral sclerosis: the role of oxidative stress and altered homeostasis of metals. AB - Amyotrophic lateral sclerosis is one of the most common neurodegenerative disorders, with an incidence of about 1/100,000. One of the typical features of this progressive, lethal disease, occurring both sporadically and as a familial disorder, is degeneration of cortical and spinal motor neurones. Present evidence indicates that loss of neurones in patients results from a complex interplay among oxidative injury, excitotoxic stimulation, dysfunction of critical proteins and genetic factors. This review focuses on existing evidence that oxidative stress is a major culprit in the pathogenesis of amyotrophic lateral sclerosis. An increase in reactive oxygen species and in products of oxidation has been observed both in post-mortem samples and in experimental models for ALS. This increase may be consequent to altered metabolism of copper and iron ions, that share the property to undergo redox cycling and generate reactive oxygen species. Metal-mediated oxidative stress would lead to several intracellular alterations and contribute to the induction of cell death pathways. PMID- 12909280 TI - Chronic lithium chloride treatment has variable effects on motor behaviour and survival of mice transgenic for the Huntington's disease mutation. AB - Expression of the Huntington's disease (HD) mutation in mice (R6/2 line) causes a progressive neurological phenotype that includes deterioration of motor function resembling that seen in HD. The current study sought to determine whether or not chronic treatment of R6/2 mice with lithium chloride would have an effect on the progression of the phenotype, in light of lithium's reported neuroprotective and anti-depressive properties. Treatment began either before or after the onset of symptoms. Chronic treatment with lithium caused a significant improvement in rotarod performance when treatment was started post- but not pre-symptomatically. There was no overall effect on survival in either group, but further analysis revealed that in the post-symptomatic group, mice could be assigned to one of two distinct groups, depending on the effects of lithium. One subgroup of mice lost weight faster, died earlier and showed rotarod performance similar to the vehicle treated controls. The other subgroup lost weight at a normal rate, died at a similar age, but showed greatly improved motor performance compared to controls. The improvement in rotarod performance suggests that lithium may improve motor symptoms as well as depression in some HD patients. PMID- 12909281 TI - Switch time-point for rapid experience-dependent changes in zinc-containing circuits in the mouse barrel cortex. AB - It has been previously demonstrated that in the mouse barrel cortex, synaptic zinc is regulated by sensory experience. In adult mice, cutting selected vibrissae produced a rapid but transient elevation of synaptic zinc in the corresponding barrels several hours later, whereas in 8 day-old animals this procedure did not affect synaptic zinc. In the present study, we wished to determine the postnatal age at which zinc-containing terminals gain the ability to respond rapidly to a restriction of sensory input. We therefore examined the effects of 1-day sensory deprivation starting at different postnatal ages. For this purpose we unilaterally trimmed all rows of vibrissae, except for row C, and we then visualized synaptic zinc in the barrel cortex 24h later. Up to postnatal day 15 such procedure had no effect on the level of synaptic zinc. However, beginning at postnatal day 16, 1-day sensory deprivation produced an increase in synaptic zinc within hollows of deprived rows of barrels as compared to non deprived rows. These results show that during development there is a specific time-point after which zinc-containing circuits may respond rapidly to altered sensory inputs. A comparison of these findings with previous results obtained after chronic sensory deprivation suggests that a specific time window exists in development for persistent alterations in zinc-containing circuits. PMID- 12909282 TI - The effects of certain H(1)-antagonists on visual evoked potential in rats. AB - The present study was undertaken to clarify the effects of certain H(1) antagonists on visual evoked potential (VEP) in rats. Pyrilamine (5 and 10 mg/kg), diphenhydramine (5 and 10 mg/kg) and chlorpheniramine (10 and 20 mg/kg) caused a significant reduction in the amplitude of late VEP components (P(3) N(3), N(3)-P(4)), although these drugs showed no significant changes in early VEP components (P(1)-N(1), N(1)-P(2)). Cyproheptadine caused a slight enhancement of late components of VEP at a dose of 20 mg/kg. On the other hand, epinastine caused no significant effect on late VEP components even at a dose of 20 mg/kg. The reduction in the late VEP components induced by pyrilamine and diphenhydramine was significantly antagonised by pre-treatment of histidine (200 and 500 mg/kg), but not by physostigmine even at a dose of 0.01 mg/kg. The effect induced by cyproheptadine was significantly potentiated by histidine (500 mg/kg), and significantly reduced by DOI (2 mg/kg). These results indicate that an inhibition of the late VEP components induced by H(1)-antagonist pyrilamine, diphenhydramine and chlorpheniramine may be due to an inhibition of specific sensory system relating the histaminergic mechanisms. In addition, slight enhancement of these components induced by cyproheptadine may be attributable to its anti-serotonergic effects. PMID- 12909283 TI - Effects of delayed administration of (-)-epigallocatechin gallate, a green tea polyphenol on the changes in polyamine levels and neuronal damage after transient forebrain ischemia in gerbils. AB - (-)-Epigallocatechin gallate has a potent antioxidant property and can reduce free radical-induced lipid peroxidation as a green tea polyphenol. In previous study, systemic administration of (-)-epigallocatechin gallate immediately after ischemia has been shown to inhibit the hippocampal neuronal damage in the gerbil model of global ischemia. Polyamines are thought to be important in the generation of brain edema and neuronal cell damage associated with various types of excitatory neurotoxicity. We examined the effects of delayed administration of (-)-epigallocatechin gallate on the changes in polyamine levels and neuronal damage after transient global ischemia in gerbils. To produce transient global ischemia, both common carotid arteries were occluded for 3 min with micro-clips. The gerbils were treated with (-)-epigallocatechin gallate (50 mg/kg, i.p.) at 1 or 3 h after ischemia. The polyamines; putrescine, spermidine, and spermine levels were examined using high performance liquid chromatography in the cerebral cortex and hippocampus 24 h after ischemia. Putrescine levels in the cerebral cortex and hippocampus were increased significantly after ischemia and the delayed administrations of (-)-epigallocatechin gallate (1 or 3 h after ischemia) attenuated the increases. Only minor changes were noted in the spermidine and spermine levels after ischemia. In histology, neuronal injuries in the hippocampal CA1 regions were evaluated quantitatively 5 days after ischemia. (-) Epigallocatechin gallate administered 1 h or 3 after ischemia significantly reduced hippocampal neuronal damage. The present results show that the delayed administrations of (-)-epigallocatechin gallate inhibit the transient global ischemia-induced increase of putrescine levels in the cerebral cortex and hippocampus. (-)-Epigallocatechin gallate is neuroprotective against neuronal damage even when administered up to 3 h after global ischemia. These findings suggest that (-)-epigallocatechin gallate may be promising in the acute treatment of stroke. PMID- 12909284 TI - Role of aging and striatal nitric oxide synthase activity in an animal model of tardive dyskinesia. AB - The risk of tardive dyskinesia (TD) increases with advancing age. Haloperidol increases striatal oxidative stress and inhibits nitric oxide (NO) synthase (NOS) in vitro. Biological aging is associated with increased oxidative stress and reduced brain NOS activity. This paper has explored aging and striatal NOS activity ex vivo as co-morbid factors in an animal model of TD. Young adult, mature adult and aged rats were treated with water or haloperidol (1.5 mg/kg per day) for 12 weeks. Vacous chewing movements (VCM) were monitored, as was striatal NOS activity. Aging significantly increased spontaneous VCM in mature and aged animals and progressively attenuated NOS activity in both mature adult and aged rats compared to young animals, and numerically lower in aged versus mature adult animals. Haloperidol significantly increased VCM in all age groups, while significantly reducing NOS activity in young and mature adults but not aged. Reduced NOS activity after haloperidol treatment was significantly lower in mature compared to young rats, but only numerically lower in aged rats receiving the drug, with a slight increase noted in the latter. In the current model, aging did not markedly alter haloperidol-induced VCM. Abrogated striatal nitrergic activity, therefore, underlies aging and haloperidol-induced VCM. Compensatory nitrergic mechanisms may preclude progressive NOS suppression and dyskinesia under conditions of advanced age and NOS inhibition. PMID- 12909285 TI - Effect of the inhibition of serotonin biosynthesis on the antinociception induced by nonsteroidal anti-inflammatory drugs. AB - The antinociceptive activity of nonsteroidal anti-inflammatory drugs (NSAIDs) has been explained mainly on the basis of their inhibition of the enzyme cyclooxygenase (COX); however, this inhibition is not enough to completely explain the analgesic efficacy of these drugs. The modulation exerted by serotonergic systems on antinociception is well known. The purpose of the present work was to further explore the role of serotonin in the antinociceptive activity of NSAIDs using the writhing test and the tail-flick test of the mice after the inhibition of serotonin biosynthesis with intraperitoneal p-chlorophenylalanine (p-CPA). Pretreatment with p-CPA produced a significant decrease in the antinociceptive activity of NSAIDs administered either by the intraperitoneal or intrathecal routes, in both algesiometric tests. These results suggest a complementary mechanism of antinociception for NSAIDs, independent of their ability to inhibit the activity of COX, involving the activation of descending serotonergic pathways. By the pharmacological nature of the study, one limitation was the absence of biochemical measurement of the synthesis of 5-HT, since the reduction of the brain 5-HT synthesis by pretreatment with p-CPA will be expressed as a diminished antinociceptive activity of NSAIDs, which would be a new argument to consider NSAIDs acting as central analgesic agents. PMID- 12909286 TI - Expression of glutamate transporter, GABRA6, serine proteinase inhibitor 2 and low levels of glutamate and GABA in the brain of knock-out mouse for Canavan disease. AB - Canavan disease (CD) is an autosomal recessive leukodystrophy characterized by spongy degeneration of the brain. The clinical features of CD are hypotonia, megalencephaly, and mental retardation leading to early death. While aspartoacylase (ASPA) activity increases with age in the wild type mouse brain, there is no ASPA activity in the CD mouse brain. So far ASPA deficiency and elevated NAA have been ascribed with the CD. Other factors affecting the brain that result from ASPA deficiency may lead pathophysiology of CD. The NMR spectra and amino acid analysis showed lower levels of glutamate and gamma-aminobutyric acid in the CD mouse brain compared to the wild type. Microarray gene expression on CD mouse brain showed glutamate transporter-EAAT4 and gamma-aminobutyric acid A receptor, subunit alpha6 (GABRA6) were lower 9.7- and 119.1-fold, respectively. Serine proteinase inhibitor 2 (Spi2) was 29.9-fold higher in the CD mouse brain compared to the wild type. The decrease of GABRA6 and high expression of Spi2 in CD mouse brain were also confirmed by real-time RT-PCR. This first report showing abnormal expression of EAAT4, GABRA6, Spi2 combined with lower levels of glutamate and GABA are likely to be associated with the pathophysiology of CD. PMID- 12909287 TI - Perinatal exposure to aluminum alters neuronal nitric oxide synthase expression in the frontal cortex of rat offspring. AB - Disturbance of the neuronal nitric oxide signaling pathway by chronic exposure to aluminum (Al) in drinking water may be a causal factor of neurological disorders in offspring. In order to investigate the relationship between Al administration and expression of neuronal nitric oxide synthase (nNOS), the numbers and distribution patterns of nNOS-immunoreactive neurons were examined in the frontal cortex of offspring after exposure to 0, 5, and 10 mM of Al in drinking water during prenatal and neonatal periods. At the bregma 0.20 level, the number of nNOS-positive neurons was significantly increased (10%) and decreased (17%) following exposure to 5 and 10 mM of Al in drinking water, respectively. The change was more severe in the upper layer than in deep layer of the cortex. In contrast, at the bregma -2.80 level, the number and distribution pattern was not significantly changed following exposure to Al. These data suggest that Al toxicity may be mediated through disturbances to the nitric oxide signaling pathway and exhibits a biphasic effect, especially in the frontal area of the cortex. In addition, the results suggest that impaired expression of nNOS plays an important role in the development of neurological syndrome caused by an exposure to Al during the early developmental stage. PMID- 12909288 TI - Role of the central amygdaloid nucleus in shaping the discharge of gustatory neurons in the rat parabrachial nucleus. AB - The central amygdaloid nucleus (CeA) receives projection from the parabrachial nucleus (PBN) gustatory neurons and descendingly projects to the PBN. To assess if the CeA is involved in modulating the activity of gustatory neurons in the PBN, the effects of electrical stimulation and electrolytic lesion of CeA on PBN gustatory neurons were observed. Of 60 neurons observed, 30 were classified as NaCl-best, 18 as HCl-best, 5 as Quinine HCl (QHCl)-best, and 7 as sucrose-best. During CeA stimulation, the responses to at least one effective stimulus were inhibited in most PBN neurons, with the response magnitudes to HCl and QHCl significantly decreased (P<0.01). In contrast, bilateral lesions of CeA facilitated the responses to HCl and QHCl (P<0.01). According to the best stimulus category, the effects on the responses to HCl and QHCl were similarly subjected to these modulations either during electrical stimulation or after electrolytic lesions of CeA. Analyses of across-unit patterns indicated that the CeA stimulation increased the chemical selection of PBN taste neurons while the CeA lesions depressed the effect on the chemical selection between NaCl and QHCl. These findings suggest that the CeA may be involved in mediating feeding behavior via modulating the activity of gustatory neurons of PBN. PMID- 12909289 TI - Hydroxyl radicals release in rat striatum involves metabotropic glutamate receptors. AB - Disruption of glutamate homeostasis frequently leads to oxidative stress and to the release of hydroxyl radicals (radical OH). Here, we investigated, via a microdialysis approach, the possible involvement of metabotropic glutamate receptors in the glutamate-induced release of hydroxyl radicals in adult rat striatum. Glutamate was applied at low amount, resulting in a moderate release that was not inhibited by dizocilpine (MK-801), a specific NMDA receptor antagonist. (RS)-alpha-methyl-4-carboxyphenylglycine (MCPG), a broad spectrum metabotropic antagonist, that does not exert any effect on the basal release of radical OH suppressed their response to glutamate. (+/-)-1-aminocyclopentane trans-1,3-dicarboxylic acid (t-ACPD), a non-selective metabotropic glutamate receptors agonist, promoted an radical OH release almost similar to that observed after glutamate, which was similarly impaired by co-infusion with MCPG. By contrast, infusion of (RS)-3,5-dihydroxyphenylglycine (DHPG), a more specific group I metabotropic glutamate receptors agonist, did not result in any appreciable radical OH response. Thus, beside NMDA receptors, some metabotropic glutamate receptors may also be involved in the glutamate-induced release of hydroxyl radicals. PMID- 12909290 TI - Location of putative glutamatergic neurons projecting to the medial preoptic area of the rat hypothalamus. AB - The medial preoptic area is a key structure in the neural control of reproduction. Considerable evidence has accumulated indicating that glutamatergic innervation of the area plays an important role in this control. Sources of the glutamatergic input are unknown. Present investigations were aimed at studying this question. [3H]D-aspartate, which is selectively taken up by high-affinity uptake sites at presynaptic endings that use glutamate or aspartate as a transmitter, and is transported back to the cell body, was injected into the medial preoptic area. The neurons retrogradely labelled with [3H]D-aspartate were detected autoradiographically. Labelled cells were found in several telencephalic and diencephalic structures, but not in the brainstem. Within the telencephalon, labelled neurons were detected in the lateral septum, bed nucleus of the stria terminalis and amygdala. Diencephalic structures included the medial preoptic area itself, hypothalamic paraventricular, suprachiasmatic, ventromedial, arcuate, ventral premammillary, supramammillary and thalamic paraventricular nuclei. All of them are known to project to this area. The findings provide the first neuromorphological data on the location of putative glutamatergic neurons projecting to the medial preoptic area. Furthermore, they indicate that local putative glutamatergic neurons as well as several telencephalic and diencephalic structures contribute to the glutamatergic innervation of the area. PMID- 12909291 TI - How to diagnose human brain damage and inflammation. PMID- 12909292 TI - Searching for needles in haystacks-the genetics of multiple sclerosis and other common neurological diseases. AB - Recent years have witnessed considerable advances in our understanding of monogenic neurodegenerative diseases, such as hereditary motor sensory neuropathy and Huntington's Chorea. Progress has been slower in the genetic dissection of other more common neurological diseases with a complex mode of inheritance. The identification of relevant genes in some, such as Alzheimer's disease (AD) or Parkinson's disease (PD), has been facilitated by characteristic pathological findings and autosomal dominant inheritance in a proportion of early onset families. Attempts to identify relevant genes for multiple sclerosis have highlighted the role of the major histocompatibility complex, but so far failed to unequivocally implicate other immunologic or structural candidate genes. Six linkage-based whole genome screens have been completed in multiple sclerosis and several regions of interest have been identified. As technology and progress in the human genome project advance, it has become clear that future studies of common neurological diseases will depend critically on the availability of large sample sizes and will have to address issues of disease heterogeneity. PMID- 12909293 TI - CSF markers for pathogenic processes in Alzheimer's disease: diagnostic implications and use in clinical neurochemistry. AB - In view of current (acetylcholine esterase (AChE) inhibitors) and future (e.g. gamma-secretase inhibitors) therapeutic compounds for treatment of Alzheimer's disease (AD), the development and evaluation of cerebrospinal fluid (CSF) biomarkers for AD has become a rapidly growing research field. Diagnostic biomarkers for AD would be especially valuable as aids to diagnosis early in the course of the disease, when correct diagnosis is difficult, and when therapeutic compounds have the greatest potential for being effective. This paper reviews CSF biomarkers for AD, with emphasis on their role in the clinical diagnosis. The two most studied biochemical markers, CSF-tau and CSF-Abeta42, have high sensitivity to identify AD, but the specificity against other dementias is lower. The addition of phosphorylated tau (P-Tau) seems to increase the specificity for the diagnosis of AD, since normal levels are found in both frontotemporal and Lewy body dementia, and in cerebrovascular disease. These CSF markers may be useful as diagnostic aids, especially to discriminate early or incipient AD from age associated memory impairment, depression, and some secondary dementias. PMID- 12909294 TI - Advances in the development of biomarkers for Alzheimer's disease: from CSF total tau and Abeta(1-42) proteins to phosphorylated tau protein. AB - Advances have been made to establish biological markers of Alzheimer's disease (AD). Measurement of total tau (t-tau) and beta-amyloid(1-42) (Abeta(1-42)) in the cerebrospinal fluid (CSF) seems useful to discriminate early and incipient AD from age-associated memory-impairment, depression, and some secondary dementias. New immunoassays to detect different phosphorylated tau epitopes (p-tau) have recently been developed. P-tau phosphorylated at threonine 231 (p-tau(231)) showed improvements compared to t-tau in the early detection of AD in subjects with mild cognitive impairment. As p-tau(231) declined during the course of AD, it may have potential to track disease progression. Additionally, p-tau(231) improved differential diagnosis between AD, frontotemporal dementia, and geriatric major depression. P-tau phosphorylated at threonine 181 improved diagnostic accuracy between AD and dementia with Lewy bodies. P-tau phosphorylated at serine 199 demonstrated high discriminative power between AD and non-Alzheimer's dementia. P-tau phosphorylated at serine 306/serine 404 improved differential diagnosis between AD and vascular dementia. A comparative study of the different p-tau epitopes is currently under way. In summary, first clinical multi-center studies suggest that measurement of phosphorylated tau proteins may significantly improve early and differential diagnosis and may come close to fulfilling proposed criteria of a biological marker for AD. PMID- 12909295 TI - Cerebral pattern of pro- and anti-inflammatory cytokines in dementias. AB - The knowledge regarding putative inflammatory component(s) participating in Alzheimer's disease (AD) and vascular dementia (VAD) is scarce. Recently, we have demonstrated the presence of certain inflammatory cytokines in the cerebrospinal fluid (CSF) of demented patients. Although the initial event(s) triggering the neurodegenerative processes in AD versus VAD may be different and lead to different neuropathological changes, it may initiate a similar cascade of cytokine production in response to neuronal injury. The cytokines released in the central nervous system (CNS) may, in turn, act in a similar manner in both diseases, amplifying some pathological changes such as amyloidogenesis and white matter lesions or on the contrary acting as neuroprotective molecules. This review will focus on the intracerebral production of the pro- and anti inflammatory cytokines interleukin IL-1beta, IL-1 receptor antagonist (IL-1ra), IL-6 and TNF-alpha in dementia, and their relation to gene polymorphism, to cerebral neuronal damage, apoptosis, and to clinical variables of dementia. Our results, which show for the first time strikingly increased CSF levels of TNF alpha but not of TNF-beta, IL-1beta or IL-6 in AD and VAD, may form a conceptual framework for further studies of neuroprotective mechanisms in dementias. PMID- 12909296 TI - Protein S-100B, neuron-specific enolase (NSE), myelin basic protein (MBP) and glial fibrillary acidic protein (GFAP) in cerebrospinal fluid (CSF) and blood of neurological patients. AB - In this study, data about protein S-100B, neuron-specific enolase, myelin basic protein and glial fibrillary acidic protein in cerebrospinal fluid and blood of patients with an acute or chronic progressive neurological disorder with brain damage are reviewed. Especially in disorders with acute brain damage, determination of these proteins in CSF and blood can be helpful to establish structural and/or functional brain damage to determine severity and prognosis of the disease process and to monitor treatment effects. PMID- 12909297 TI - Studies of the brain specificity of S100B and neuron-specific enolase (NSE) in blood serum of acute care patients. AB - Laboratory monitoring with damage markers of brain and of non-nervous tissues in blood serum of 401 acute care patients showed increased contents of neuron specific enolase (NSE) and S100B besides raised levels of markers of heart, skeletal muscle, bile duct, liver, prostate, kidney, salivary gland damage or of inflammatory stress to varying frequencies. Correlation between raised NSE and S100B contents ascertained brain damage. Correlation between raised NSE and troponin I (cTnI) values indicated brain damage induced by heart failure (probably caused by hypoxia and anemia); this was assessed with correlations between NSE and other heart markers, e.g. creatine kinase (CK) isoenzymes, alpha hydroxybutyrate dehydrogenase. S100B did not show such correlations: data indicated S100B release from non-nervous tissues having high S100B content, e.g. fat, cartilage, skin. S100B release might be triggered by inflammatory stress and tissue damage. This was further supported by low NSE/S100B concentration ratios in serum compared to cerebrospinal fluid (CSF) of patients with comatose state, convulsive status, or intracerebral hemorrhage. Our data revealed CSF to be the relevant sample to monitor brain damage with NSE and S100B, whereas in serum raised S100B levels together with normal NSE levels indicated release from non nervous tissues of acute care patients pointing out multi-organ dysfunction. PMID- 12909298 TI - Cerebrospinal fluid (CSF) and serum S100B: release and wash-out pattern. AB - S100B is an important brain specific protein for monitoring damage and activation of astrocytes. Using a straight forward, non-resource demanding in-house ELISA technique we measured S100B in cerebrospinal fluid (CSF) and serum in patients with traumatic brain injury (TBI) (serum), subarachnoid hemorrhage (SAH) (CSF, serum), intracranial hemorrhage (ICH) (CSF, serum), normal controls (NC) (serum) and a reference population (CSF, N=409). The release and wash-out pattern found in CSF and serum are discussed in relation to the three main determinants for increased brain specific protein levels in body fluids: (i) total mass effect; (ii) pathology; and (iii) time effect. PMID- 12909299 TI - New and old diagnostic markers of meningitis in cerebrospinal fluid (CSF). AB - Five new markers (tumor necrosis factor TNF-alpha, interleukin IL-1 beta, IL-6, IL-8, lipopolysaccharide binding protein (LBP)) and 11 old classical markers were evaluated in 180 cerebrospinal fluid (CSF) and serum pairs to discriminate acute bacterial meningitis (BM) on admission from aseptic (viral) meningitis (AM), bacterial meningitis treated with antibiotics (TM) from AM, and AM from multiple sclerosis (MS). Statistical tests were computed which classified correctly > or =90% of the patients with BM, TM, AM at a sum minimum of false positive plus false negative results, and which reached additionally > or =90% sensitivity and specificity. To discriminate BM from AM, CSF IL-6 test > or =500 ng/l and CSF IL 1 beta test > or =8 ng/l besides CSF lactate test > or =3.5mM/l and CSF granulocyte test > or =150 M/l were revealed. CSF lactate test > or =3.2 mmol/l discriminated TM from AM. CSF leukocyte test > or =35 M/l discriminated AM from MS. Tests with the new markers were more laborious, expensive, and time consuming compared to CSF lactate test. Test candidates, detecting > or =80% of patients with > or =80% sensitivity and specificity, were evaluated with CSF TNF-alpha, IL 8 and LBP, serum IL-6, CSF leukocytes, lymphocytes and monocytes, Qglucose, CSF total protein, albumin, and Qalbumin. All tests should be reviewed in context of clinical findings to diagnose BM reliably. PMID- 12909300 TI - Clinical relevance of polymerase chain reaction (PCR) assays and antigen-driven immunoblots for the diagnosis of neurological infectious diseases. AB - Polymerase chain reaction assays are a powerful tool for detecting the presence of infectious genomes in the cerebrospinal fluid. Positive results always mean a current or pending infection of the central nervous system. Subacute (>7 days) or chronic infections induce an intrathecal humoral immune response and the appearance of oligoclonal IgG antibodies directed against the causal infectious agent. This local synthesis may be observed even in cases of severe systemic immunodeficiency. The use of polymerase chain reactions in combination with the detection of a specific intrathecal immune response should represent the most reliable strategy for the diagnosis of viral and chronic infections of the central nervous system. The authors describe their experience, using this approach, in herpetic encephalitis, acute and recurrent herpetic meningitis, varicella zoster-induced neurological diseases, cytomegalovirus encephalitis, progressive multifocal leukoencephalitis and tuberculous meningitis. PMID- 12909301 TI - Distinct heterogeneity of IgG immune response in cerebrospinal fluid (CSF) detected by isoelectric focusing (IEF) with extended immunofixation. AB - An optimized automated IEF procedure in polyacrylamide micro gels and immunofixation with 10 monospecific antibodies against some fragments of the IgG molecule and against the whole IgG isotypes IgG(1), IgG(2), IgG(3), IgG(4), respectively, detected oligoclonal bands (OBs) within acid, neutral, and alkaline ranges of the gels. Accuracy and reliability of the OB assay for detection of an intrathecal IgG synthesis proved to be higher with immunofixation than with silver staining of bands precipitated by trichloroacetic acid. CSF OBs were specified as lambda or kappa IgG subfractions, respectively, duplex IgGs precipitated with anti-lambda, anti-kappa, and anti-Fc antibodies at the same pI. Most of OBs were classified as belonging to either IgG(1), IgG(2), IgG(3), or IgG(4) isotypes in CSF. The IEF procedure additionally allowed the discrimination of both free light chains and possible "free" heavy gamma chain fragments in CSF, when immunofixation was done with monospecific antibodies against both light chains and gamma chain fragments (e.g. anti-F(ab')(2), anti-Fd, anti-Fc, anti C(H)2). The results pointed out a distinct heterogeneity of the IgG immune response in human CSF. The IEF procedure with extended immunofixation is recommended for completion of the basic laboratory procedures used in neuroimmunology in order to discriminate inflammatory processes in human CNS. PMID- 12909302 TI - Neuropathology of multiple sclerosis-new concepts. AB - Multiple sclerosis is a chronic inflammatory disease of the central nervous system with profound heterogeneity in clinical course, neuroradiological presentation and response to therapy. The pathological analysis of 235 actively demyelinating lesions coming from three centers revealed different structural and immunological features suggesting that different pathogenetic mechanisms are involved in lesion formation. On the basis of the presence or absence of immunoglobulin and complement deposition, myelin protein loss and the patterns of oligodendrocyte degeneration beside a T cell- and macrophage-dominated immune response, four distinct patterns of demyelination have been identified. In this short review, possible paraclinical markers for tissue destruction on the basis of the main features of myelin destruction are discussed. Furthermore, the importance of early axonal damage in multiple sclerosis is highlighted. PMID- 12909303 TI - Approach to discriminate subgroups in multiple sclerosis with cerebrospinal fluid (CSF) basic inflammation indices and TNF-alpha, IL-1beta, IL-6, IL-8. AB - Lumbar CSF and serum pairs of untreated multiple sclerosis patients (MS; n=47) were analyzed on admission. On average, higher CSF leukocyte (lymphocyte and monocyte) counts, IgG index, CSF IgG contents, but not of TNF-alpha, IL-1beta, IL 6, IL-8 in CSF and serum, were revealed in all MS or patients with long disease course (LO-MS) compared with controls. In primary progressive MS (PP-MS) cell counts were low, but IgG contents were high, when compared to relapsing-remitting MS (RR-MS). In clinically probable MS (CP-MS) both contents were low, in clinically definite MS (CD-MS) high. Spearman's correlation with the four monokines and the basic indices in CSF revealed activation patterns known for microglia/macrophages in the four MS subgroups, for astrocytes in CP-MS and RR MS, for CSF lymphocytes in CP-MS and PP-MS, for cells of blood-brain barrier (BBB) in CP-MS, for intrathecal IgG synthesis in PP-MS and for lymphocyte transfer in CD-MS. Correlations between CSF and serum parameters indicated CNS disease processes to be associated with systemic processes of inflammation (acute, chronic) in CD-MS, RR-MS, and PP-MS in different ways. CSF IgG content, IgG index and systemic markers of inflammation correlated with overall disability scores in LO-MS; increasing levels may indicate a bad outcome. PMID- 12909304 TI - Chemokines and matrix metalloproteinase-9 in leukocyte recruitment to the central nervous system. AB - Chemokines and matrix metalloproteinases (MMPs) play key roles in leukocyte migration across the blood-brain barrier (BBB) in infectious and inflammatory diseases, including multiple sclerosis (MS). In MS some chemokine receptors are expressed by an increased percentage of T cells in blood, the CSF concentration of chemokine ligands for these receptors is increased, and there is accumulation of T cells expressing relevant chemokine receptors in CSF and in the CNS parenchyma. Chemokine receptor expression patterns appear to reflect disease activity and disease stage in MS. MMPs are constitutively expressed or induced by proinflammatory cytokines and chemokines in leukocytes and CNS-resident cells. Several MMPs are expressed in MS plaques, and the CSF concentration of MMP-9 is increased in MS. The CSF concentration of MMP-9 may reflect disease activity in MS, and the CSF concentration of MMP-9 is higher in patients carrying the MS associated HLA type DRB1 1501. We review how chemokines and MMP-9 may be involved in the pathogenesis of MS by controlling leukocyte migration between different functional compartments. Measuring expression of these molecules may find use as surrogate markers of disease activity in MS, and interfering with their function holds promise as a novel therapeutic strategy in MS. PMID- 12909306 TI - Adenosine-the missing link to understanding homocysteine pathogenicity or more smoke on the horizon? PMID- 12909305 TI - Adhesion molecules and matrix metalloproteinases in Multiple Sclerosis: effects induced by Interferon-beta. AB - In Multiple Sclerosis (MS) pathology, early inflammation involves leukocyte migration across the blood-brain barrier (BBB) within the central nervous system. In this process, adhesion molecules (AMs), both membrane-bound and soluble circulating forms, and matrix metalloproteinases (MMPs) certainly play a regulatory role. In MS, recombinant Interferon-beta (rIFNbeta) is effective in reducing gadolinium contrast-enhancing lesions on magnetic resonance imaging and this suggests that it may reduce BBB damage or even restore its integrity by different mechanisms that include interference with both AM and MMP pathways. This review will highlight the effects induced by rIFNbeta, both in vitro and in vivo, on cell-bound and soluble forms of AMs and on MMPs. PMID- 12909307 TI - Negative inotropy starts with the beta(3)-adrenoceptor. PMID- 12909308 TI - Mitochondria as common endpoints in early and late preconditioning. PMID- 12909309 TI - Ischemia/reperfusion-induced apoptosis: connecting nitric oxide and cell cycle regulators. PMID- 12909310 TI - Potential role for adenosine in the pathogenesis of the vascular complications of hyperhomocysteinemia. AB - Hyperhomocysteinemia is an independent risk factor for cardiovascular disease. Most previous investigations focused on the role of homocysteine as direct pathogenetic factor for these adverse vascular events. However, the exact pathophysiological mechanism is still unknown. In this review we discuss the hypothesis that a decreased extracellular concentration of adenosine could contribute to the adverse cardiovascular effects of hyperhomocysteinemia. Fundamental to this hypothesis is that, in vivo, any increase in the plasma concentration of homocysteine reflects an increased intracellular homocysteine concentration, which inevitably will result in a decrease in the adenosine concentration. In this situation, the hydrolase reaction catalysed by S adenosylhomocysteine hydrolase will reverse and S-adenosylhomocysteine will accumulate at the expense of adenosine. Stimulation of adenosine receptors by adenosine results in various cardio- and vasoprotective actions, like modulation of vascular resistance, presynaptic inhibition of norepinephrine release, ischaemic preconditioning, inhibition of platelet aggregation, modulation of inflammation and regulation of vascular cell proliferation and death. In this respect, a decrease in the adenosine concentration could contribute significantly to the cardiovascular effects of hyperhomocysteinemia. PMID- 12909311 TI - Microparticles in cardiovascular diseases. AB - Microparticles are membrane vesicles released from many different cell types. There are two mechanisms that can result in their formation, cell activation and apoptosis. In these two mechanisms, different pathways are involved in microparticle generation. Microparticle generation seems to be a well regulated process. Microparticles vary in size, phospholipid and protein composition. They have a potent pro-inflammatory effect, promote coagulation and affect vascular function. Since these processes are all involved in the pathogenesis of cardiovascular disease and circulating microparticle numbers are altered in many cardiovascular diseases, a role for microparticles in the pathogenesis of cardiovascular diseases is likely. Although hard evidence for a role of microparticles in cardiovascular diseases at present is still only limited, new evidence is accumulating rapidly to support this theory. Elucidation of the microparticle composition and the mechanisms involved in exertion of their effects will supply this evidence and enable us to develop additional intervention strategies for prevention and treatment of cardiovascular diseases. PMID- 12909312 TI - beta3-Adrenergic stimulation produces a decrease of cardiac contractility ex vivo in mice overexpressing the human beta3-adrenergic receptor. AB - OBJECTIVES: The regulation of cardiac function by catecholamines involves three populations of beta-adrenoceptor (beta-AR). beta(1)- and beta(2)-AR stimulations produce an increase in contractility and beta(3)-AR stimulation mediates a negative inotropic effect in human ventricular muscle. Because of the lack of suitable animal models, we have generated transgenic mice with cardiac-specific expression of the human beta(3)-AR (TG beta(3) mice). METHODS: TG beta(3) mice were produced by microinjection of the human beta(3)-AR under the control of the alpha myosin heavy chain promoter. Phenotypic analyses comprised beta(3)-AR mRNA and protein determinations, histological studies, electrocardiogram, contractility and cyclic nucleotide measurements. RESULTS: TG beta(3) mice presented no histological evidence of myocyte hypertrophy or fibrogenesis. In basal conditions, TG beta(3) mice were characterized by an increase in heart rate and an acceleration of twitch parameters without modification of its amplitude. beta(3)-AR agonists (CL 316243, SR 58611A) decreased contractility at low concentrations (1-100 nM). At high concentrations, the negative inotropic effect was abolished. Pretreatment with nadolol, a beta(1)/beta(2)-AR blocker, blunted the rebound in peak tension elicited by beta(3)-AR agonists suggesting a non specific action of these compounds on beta(1)- and beta(2)-AR. The involvement of beta(3)-AR in the negative inotropic effect was confirmed by the pretreatment with bupranolol, a non-selective beta-AR antagonist, which fully abolished the effects of SR 58611A. The negative inotropic effect was associated with an increase in intracellular cGMP level. CONCLUSIONS: We conclude that cardiac overexpression of beta(3)-AR in mice reproduces ex vivo the negative inotropic effects obtained with beta(3)-AR stimulation in human ventricular tissues. PMID- 12909313 TI - Second window of ischemic preconditioning regulates mitochondrial permeability transition pore by enhancing Bcl-2 expression. AB - OBJECTIVE: The second window of protection (SWOP) following brief coronary artery occlusion begins at 24 h and may last up to 72 h and occurs via many unknown mechanisms. We investigated the role of the mitochondrial permeability transition pore (PTP), a non specific pore in the inner membrane of the mitochondria in this phenomenon. METHODS: Ischemic preconditioning (IP) was induced in Wistar rats by left coronary artery occlusion (four, 3-min episodes separated by 10 min of reperfusion) on day 1. On day 2, ischemia was induced with 30 min of ischemia and 120 min of reperfusion in IP and control rats. RESULTS: IP rats showed decreased myocardial infarction (MI) area vs. non-IP control rats (15.32 vs. 45.6%). Furthermore, IP rats had preserved cardiac function (heart rate, rate pressure product, coronary flow and aortic flow) and myocardial ATP (P<0.03), decreased tissue water content (73.2 vs. 90.6%), increased expression of Bcl-2, and less mitochondrial swelling, cytochrome C release and apoptosis (2.6 vs. 12.4%) when compared to sham-operated rats. Activation of the permeability transition pore with PTP activators lonidamine (10 mg/kg body weight) or atractyloside (5 mg/kg body weight) before the sustained ischemia on day 2 resulted in complete abolition of SWOP-mediated cytoprotective effects. These agents had no effect on the cytoprotective effects that took place during the first window of preconditioning. CONCLUSION: The cytoprotective effects of SWOP are dependent on PTP activation state and may involve upregulation of Bcl-2 expression. PMID- 12909314 TI - Nitric oxide inhibits ischemia/reperfusion-induced myocardial apoptosis by modulating cyclin A-associated kinase activity. AB - OBJECTIVE: Ischemia/reperfusion in the heart causes myocardial apoptosis and increase nitric oxide (NO) production. We have reported that myocardial apoptosis is related to activation of cell cycle regulatory proteins. However, the role of nitric oxide (NO) in ischemia/reperfusion-induced apoptosis is still unclear. This study was designated to elucidate novel apoptosis mechanisms induced by ischemia/reperfusion, especially the interaction between NO and cell cycle regulators. METHODS AND RESULTS: Neonatal cardiomyocytes from 1- or 2-day-old Wistar rats were subjected to 1-h ischemia and then to reperfusion. The rate of cardiomyocyte apoptosis increased significantly after 24 h of reperfusion as evaluated by TUNEL analysis. NO increased 1.8-fold after 15 min of reperfusion in cardiomyocytes. After 36 h of reperfusion, the apoptosis rate was greatly increased by the NO synthetase inhibitor, Nitro-L-arginine methyl ester (L-NAME), and decreased by the NO donor of S-nitroso-N-acetylpenicillamine (SNAP). Immunoblot analysis showed that the protein levels of cyclin A accumulated in a time-dependent manner in response to ischemia/reperfusion, and L-NAME inhibited this response. Ischemia/reperfusion also increased the activity of cyclin A associated kinase, and the apoptosis was inhibited by infection of dominant negative cdk2 adenovirus. To clarify the involvement of p21(cip1/waf1) protein, which is the suppressor of cyclin A-associated kinase, we performed immunoblot analysis and examined its kinase activity. Treatment of cardiomyocytes with L NAME suppressed the p21(cip1/waf1) protein level and increased the cyclin A associated kinase activity. The addition of SNAP showed inverse results. CONCLUSION: Our data indicates that NO released from cardiomyocytes under condition of ischemia/reperfusion exerts an antiapoptotic effect by modulating cyclin A-associated kinase activity via p21(cip1/waf1) accumulation. PMID- 12909315 TI - Andersen mutations of KCNJ2 suppress the native inward rectifier current IK1 in a dominant-negative fashion. AB - OBJECTIVE: The Andersen's syndrome is a hereditary disease, which is characterized by cardiac arrhythmias, periodic paralysis and dysmorphic features. Recently, mutations of the KCNJ2 gene, which encodes the inward rectifying potassium channel subunit Kir2.1, have been identified in affected individuals. However, the functional effects of these mutations have not yet been fully elucidated. METHODS AND RESULTS: To clarify this situation we generated known Andersen disease mutants of KCNJ2 which did not yield any measurable K(+) currents in CHO cells indicating that the Andersen mutants failed to form functional homomultimeric complexes. EGFP-tagged KCNJ2 wild-type and mutant channels distributed in a similar homogeneous pattern in the cell membrane suggesting that protein trafficking was not altered by the Andersen mutations but rather implicating that the mutations rendered the KCNJ2 channel non-functional. In heterologous coexpression experiments the Andersen mutants exerted a dominant negative effect on wild-type KCNJ2. However, the extent of suppression varied between the different KCNJ2 mutants. Given our results in CHO cells, we expressed the disease mutant KCNJ2-S136F in neonate rat cardiomyocytes using adenoviral gene transfer to test the effect of Andersen mutants on native I(K1). I(K1) density was indeed significantly reduced in KCNJ2-S136F-infected cells (n=9) compared to control cells (n=9) over a voltage range from -70 to -150 mV (P<0.05). CONCLUSION: These results support that Kir2.x channels are a critical component of native I(K1) in neonate rat cardiomyocytes and that a dominant negative suppression of I(K1) in native cells is the pathophysiological correlate of the Andersen's syndrome. PMID- 12909316 TI - Barium block of Kir2 and human cardiac inward rectifier currents: evidence for subunit-heteromeric contribution to native currents. AB - BACKGROUND: Kir2 subunits are believed to underlie the cardiac inwardly rectifying current I(K1). The subunit composition of native I(K1) currents is uncertain, and it has been suggested that heteromultimer formation may play a role. METHODS: We studied Ba(2+) block of homo- and heteromeric Kir2 channels in Xenopus oocytes and compared the properties observed to those of human cardiac I(K1) in cells isolated from myocardial biopsies of normal human hearts. RESULTS: Homomeric expression of Kir2.1 and Kir2.3 produced currents with similar Ba(2+) sensitivities (e.g. IC(50) at -120 mV: 16.2+/-3.4, n=11 and 18.5+/-2.1, n=10, respectively), but these were less sensitive to Ba(2+) than native I(K1) (4.7+/ 0.5 microM, n=10, P=0.001, P<0.001, respectively) and had different Ba(2+) blocking kinetics from cardiac I(K1). Kir2.2 sensitivity was similar to cardiac I(K1) (e.g., 2.8+/-0.4 microM, Kir2.2, n=9, vs. 4.7+/-0.5 microM for I(K1)), but the blocking kinetics of Kir2.2 were faster than those of I(K1). Currents resulting from co-expression of Kir2 subunits had similar Ba(2+) sensitivities and blocking kinetics among groups and were similar to I(K1) in both Ba(2+) sensitivity (e.g., IC(50) at -120 mV: 4.5+/-1.0, 2.5+/-0.5, and 2.3+/-0.4 microM for co-injected Kir2.1/2.2, n=6, Kir2.1/2.3, n=5, and Kir2.2/2.3, n=4, respectively) and blocking kinetics. CONCLUSION: Co-injection of Kir2 subunits results in currents with Ba(2+) blocking properties different from homomeric Kir2 expression but similar to cardiac I(K1). These observations suggest that a substantial proportion of native I(K1) may result from heteromultimer formation among diverse Kir2 family subunits. PMID- 12909317 TI - Developmental changes in IKr and IKs contribute to age-related expression of dofetilide effects on repolarization and proarrhythmia. AB - OBJECTIVE: Clinical and experimental studies suggest that immature hearts are as or more sensitive than adult hearts to adverse effects of I(Kr) blocking drugs. We hypothesized that age-dependent changes in I(Kr) and I(Ks) contribute to the different repolarization reserves and proarrhythmic effects of I(Kr) blockers in the young and adult heart. METHODS: Dogs aged 1-150 days and adults were used to study (1) proarrhythmic effects in situ of the I(Kr) blocker dofetilide; (2) dofetilide effects on action potential duration (APD) recorded with microelectrodes from left ventricular (LV) slabs; (3) I(Kr) and I(Ks) in single LV myocytes using whole-cell voltage clamp. RESULTS: In situ, dofetilide-induced proarrhythmia occurred in 40% of adults, 86% of young (20-150 day) dogs and 0% of neonatal (1-19 day) dogs (P<0.05). Isolated tissue experiments showed no transmural gradient for repolarization from neonate through 3 months of age, after which the gradient increased through adulthood. In the presence of dofetilide, the greatest APD prolongation occurred in neonates. Yet, transmural dispersion did not increase in neonates but significantly increased in young and adults. Dofetilide-induced early after depolarization (EAD) incidence was 23% in adults, 59% in young and 8% in neonates (P<0.05). I(Kr) but not I(Ks) was expressed at <30 days, whereas both currents were present in adult myocardium. CONCLUSIONS: Our data suggest that a lack of I(Ks) results in a greater dependence on I(Kr) for repolarization in neonates and is associated with exaggerated effects of I(Kr)-blockade on APD. However, APD prolongation alone is insufficient for expression of proarrhythmia, which also requires transmural dispersion of repolarization and EADs. The extent to which APD prolongation, transmural dispersion and EADs are manifested at various ages in the absence and presence of I(Kr) blocking drugs appears to be the ultimate determinant of proarrhythmia. PMID- 12909318 TI - Asynchronous development of electrical remodeling and cardiac hypertrophy in the complete AV block dog. AB - OBJECTIVE: Left ventricular hypertrophy has been associated with the prolongation of QT-time, and an increased risk of ventricular arrhythmias. The renin angiotensin system has been implicated in the development of ventricular hypertrophy. At 5 weeks complete AV block (CAVB) in the dog, there is: (1) biventricular hypertrophy associated with a transient activation of components of the renin angiotensin system, (2) increased APD, more pronounced in the left than in the right ventricle leading to spatial dispersion of repolarization, and (3) enhanced susceptibility to drug-induced torsade de pointes arrhythmias. To investigate whether these remodeling processes develop in parallel, time dependency was assessed in absence or presence of the AT1 receptor-blocker Irbesartan. METHODS AND RESULTS: Dogs in sinus rhythm, 2 and 5 weeks CAVB were compared to dogs chronically treated with Irbesartan (30 mg/kg BID). Endocardial monophasic APD of left and right ventricle was measured and susceptibility to torsade de pointes was tested by infusing Dofetilide (0.025 mg/kg/5'). Hypertrophy was determined by relating heart-to-body weight at sacrifice. Left ventricular APD had increased more than right ventricular APD at 2 and 5 weeks CAVB, leading to an increase in spatial dispersion. At that time torsade de pointes were evocable in the majority of the dogs. Hypertrophy had only developed completely at 5 weeks CAVB. Irbesartan had no effect on electrical and structural parameters or on arrhythmogenicity. CONCLUSIONS: In the CAVB dog ventricular hypertrophy is not a prerequisite for electrical remodeling or drug-induced torsade de pointes, and the AT1-receptor has no dominant role in the completion of these remodeling processes. PMID- 12909319 TI - Diverse regulation of atrial natriuretic peptide secretion by serotonin receptor subtypes. AB - OBJECTIVE: Serotonin (5-hydroxytryptamine [5-HT]) receptors are located in peripheral tissues as well as in the central nervous system. Serotonin receptors mediate positive inotropic and chronotropic effects in atria. The aim of this study was to investigate physiological role of endogenous serotonin on the regulation of atrial natriuretic peptide (ANP) secretion from the atria. METHODS: An isolated perfused nonbeating rat atrial model was used. Changes in atrial volume induced by increasing intra-atrial pressure were measured. The concentration of ANP was measured by radioimmunoassay and the translocation of ECF was measured by [3H]-inulin clearance. RESULTS: Serotonin, an endogenous 5-HT receptor agonist, caused concentration-dependent suppressions of stretch-induced ANP secretion, which were less pronounced than those caused by alpha-methyl-5-HT maleate, a 5-HT(2) receptor selective agonist. The suppression of stretch-induced ANP secretion due to serotonin and alpha-methyl-5-HT maleate was attenuated by ketanserin, a 5-HT(2) receptor antagonist, and accentuated by RS23597-190, a 5 HT(4) receptor antagonist. The suppressive effect of serotonin on ANP secretion was attenuated by neomycin, staurosporine, and chelerythrine. In contrast, 2-[1 (4-piperonyl)piperazinyl]benzothiazole, a 5-HT(4) receptor selective agonist, caused an accentuation of stretch-induced ANP secretion, which was completely blocked by RS23597-190 and SB203186 HCl but not by ketanserin. This effect was not affected by MDL12330, KT-5720, or H-89. The intracellular Ca(2+) concentration in single atrial myocytes was not changed by serotonin and agonist for either 5-HT(2) or 5-HT(4) receptor. CONCLUSIONS: These results suggest that atrial 5-HT(2) and 5-HT(4) receptor agonists have opposite actions on the regulation of ANP secretion and the suppressive effect of serotonin on the ANP secretion may act through 5-HT(2) receptor and phospholipase C pathway. PMID- 12909320 TI - Impaired relaxation in transgenic mice overexpressing junctin. AB - OBJECTIVE: Junctin is a major transmembrane protein in cardiac junctional sarcoplasmic reticulum, which forms a quaternary complex with the ryanodine receptor (Ca(2+) release channel), triadin, and calsequestrin. METHODS: To better understand the role of junctin in excitation-contraction coupling in the heart, we generated transgenic mice with targeted overexpression of junctin to mouse heart, using the alpha-MHC promoter to drive protein expression. RESULTS: The protein was overexpressed 10-fold in mouse ventricles and overexpression was accompanied by cardiac hypertrophy (19%). The levels of two other junctional SR proteins, the ryanodine receptor and triadin, were reduced by 32% and 23%, respectively. However, [3H]ryanodine binding and the expression levels of calsequestrin, phospholamban and SERCA2a remained unchanged. Cardiomyocytes from junctin-overexpressing mice exhibited impaired relaxation: Ca(2+) transients decayed at a slower rate and cell relengthening was prolonged. Isolated electrically stimulated papillary muscles from junctin-overexpressing hearts exhibited prolonged mechanical relaxation, and echocardiographic parameters of relaxation were prolonged in the living transgenic mice. The amplitude of caffeine-induced Ca(2+) transients was lower in cardiomyocytes from junctin overexpressing mice. The inactivation kinetics of L-type Ca(2+) channel were prolonged in junctin-overexpressing cardiomyocytes using Ca(2+) or Ba(2+) as charge carriers. CONCLUSION: Our data provide evidence that cardiac-specific overexpression of junctin is accompanied by impaired myocardial relaxation with prolonged Ca(2+) transient kinetics on the cardiomyocyte level. PMID- 12909321 TI - Transgenic rat hearts overexpressing SERCA2a show improved contractility under baseline conditions and pressure overload. AB - OBJECTIVE: The activity of sarcoplasmic reticulum Ca(2+)-ATPase (SERCA) is reduced in the failing myocardium. Therefore, transfer of SERCA2a cDNA is considered as a therapeutical approach. The aim of this study was analysis of the long-term effect of SERCA2a overexpression in normal as well as pressure overload challenged myocardium of transgenic rats. METHODS: Independent transgenic rat lines were established expressing the rat SERCA2a cDNA specifically in the myocardium resulting in increased SERCA2a protein levels by 30-70%. Simultaneous measurements of isometric contraction and calcium transients were carried out in right ventricular papillary muscle preparations. Hemodynamic parameters were measured in hearts of unchallenged rats as well as 10 weeks after pressure overload induced by abdominal aortic banding. RESULTS: Analysis of calcium handling and contractile parameters in isolated right ventricular papillary muscles revealed significant shortening of intracellular calcium transients and half maximal relaxation times (RT(50)). Assessing myocardial contractility in working heart preparations, both transgenic rat lines revealed elevated left ventricular pressure, improved systolic and diastolic parameters, attenuated negative force-frequency relation, and a dose-dependent beta-adrenergic effect. Aortic banding resulted in reduction of left ventricular pressure and worsening of contraction and relaxation parameters with no differences in mortality in both transgenic (+dP/dt 3084+/-96 vs. 3938+/-250 mmHg/s; RT(50) 47.0+/-1.2 vs. 36.7+/ 1.4 ms) and wild-type rats (+dP/dt 2695+/-86 vs. 3297+/-122 mmHg/s; RT(50) 53.0+/ 1.6 vs. 44.1+/-1.4). SERCA2a overexpressing hearts revealed improved hemodynamic parameters compared to wild-type controls. Acceleration of isovolumetric relaxation characterized by the index Tau was directly correlated to SERCA2a protein concentrations. CONCLUSION: Overexpression of SERCA2a protein results in a positive inotropic effect under baseline conditions remaining preserved under pressure overload without affecting mortality. Therefore therapeutic transfer of SERCA2a may become a potential approach for gene therapy of congestive heart failure. Moreover, transgenic SERCA2a rats will be useful for studies of long term SERCA2a overexpression in further cardiovascular disease models. PMID- 12909322 TI - Dynamic regulation of MEK/Erks and Akt/GSK-3beta in human end-stage heart failure after left ventricular mechanical support: myocardial mechanotransduction sensitivity as a possible molecular mechanism. AB - OBJECTIVE: Left ventricular assist devices (LVAD) are used to 'bridge' patients with end-stage heart failure to transplantation. After long-term LVAD support, ventricular function may partially recover, a process called 'reverse remodeling'. As several kinase-mediated signal transduction pathways have been implicated in the development of cardiac hypertrophy and failure, we examined the activities of the Erks, MEKs, Akt, GSK-3 beta, p70S6K, JNKs and p38 under LVAD support as well as during single myocyte strain and whole heart stretch. METHODS: Western blotting and immunohistochemistry were performed using phospho-specific antibodies in matched samples from ten patients with end-stage heart failure before and after LVAD. Cyclic strain was performed in rat neonatal cardiac myocytes, and tensile stretch applied to Langendorff-perfused mouse hearts via a left ventricular balloon. RESULTS: The activity of Erks and Akt in failing hearts dramatically decreased after LVAD support, while that of GSK-3 beta increased. There was an endo/epicardial gradient for Erk activity which persisted after LVAD despite the reduction of total Erk activity. TUNEL-positivity and myocyte size decreased after LVAD, but independently of changes in kinase activity. In cardiomyocytes and Langendorff-perfused mouse hearts both strain/stretch and its relief regulated the activities of Erks, Akt, and GSK-3 beta. CONCLUSION: Erks and Akt/GSK-3 beta are highly responsive to myocyte stretch in vitro and in vivo, and may be sensitive molecular parameters of 'reverse remodeling' under LVAD support. PMID- 12909323 TI - Expression profiling of human idiopathic dilated cardiomyopathy. AB - OBJECTIVE: To investigate the global changes accompanying human dilated cardiomyopathy (DCM) we performed a large-scale expression screen using myocardial biopsies from a group of DCM patients with moderate heart failure. By hierarchical clustering and functional annotation of the deregulated genes we examined extensive changes in the cellular and molecular processes associated to DCM. METHODS: The expression profiles were obtained using a whole genome covering library (UniGene RZPD1) comprising 30336 cDNA clones and amplified RNA from myocardiac biopsies from 10 DCM patients in comparison to tissue samples from four non-failing, healthy donors. RESULTS: By setting stringent selection criteria 364 differentially expressed, sequence-verified non-redundant transcripts were identified with a false discovery rate of <0.001. Numerous genes and ESTs were identified representing previously recognised, as well as novel DCM associated transcripts. Many of them were found to be upregulated and involved in cardiomyocyte energetics, muscle contraction or signalling. Two hundred and twenty-two deregulated transcripts were functionally annotated and hierarchically clustered providing an insight into the pathophysiology of DCM. Data was validated using the MLP-deficient mouse, in which several differentially expressed transcripts identified in the human DCM biopsies could be confirmed. CONCLUSIONS: We report the first genome-wide expression profile analysis using cardiac biopsies from DCM patients at various stages of the disease. Although there is a diversity of links between the cytoskeleton and the initiation of DCM, we speculate that genes implicated in intracellular signalling and in muscle contraction are associated with early stages of the disease. Altogether this study represents the most comprehensive and inclusive molecular portrait of human cardiomyopathy to date. PMID- 12909324 TI - Effects of endotoxic shock on right ventricular systolic function and mechanical efficiency. AB - OBJECTIVE: To investigate the effects of endotoxin infusion on right ventricular (RV) systolic function and mechanical efficiency. METHODS: Six anesthetized pigs (Endo group) received a 0.5 mg/kg endotoxin infusion over 30 min and were compared with six other anesthetized pigs (Control group) receiving placebo for 5 h. RV pressure-volume (PV) loops were obtained by the conductance catheter technique and pulmonary artery flow and pressure were measured with high-fidelity transducers. RESULTS: RV adaptation to increased afterload during the early phase of endotoxin-induced pulmonary hypertension (T30) was obtained by both homeometric and hetereometric regulations: the slope of the end-systolic PV relationship of the right ventricle increased from 1.4+/-0.2 mmHg/ml to 2.9+/-0.4 mmHg/ml (P<0.05) and RV end-diastolic volume increased from 56+/-6 ml to 64+/-11 ml (P<0.05). Consequently, right ventricular-vascular coupling was maintained at a maximum efficiency. Ninety minutes later (T120), facing the same increased afterload, the right ventricle failed to maintain its contractility to such an elevated level and, as a consequence, right ventricular-vascular uncoupling occurred. PV loop area, which is known to be highly correlated with oxygen myocardial consumption, increased from 1154+/-127 mmHg/ml (T0) to 1798+/-122 mmHg/ml (T180) (P<0.05) while RV mechanical efficiency decreased from 63+/-2% (T0) to 45+/-5% (T270) (P<0.05). CONCLUSIONS: In the very early phase of endotoxinic shock, right ventricular-vascular coupling is preserved by an increase in RV contractility. Later, myocardial oxygen consumption and energetic cost of RV contractility are increased, as evidenced by the decrease in RV efficiency, and right ventricular-vascular uncoupling occurs. Therefore, therapies aiming at restoring right ventricular-vascular coupling in endotoxic shock should attempt to increase RV contractility and to decrease RV afterload but also to preserve RV mechanical efficiency. PMID- 12909325 TI - Decrease in sarcoglycans and dystrophin in failing heart following acute myocardial infarction. AB - OBJECTIVE: Genetic defects in several sarcoglycans (SGs) and dystrophin (Dys) play a critical role in cardiomyopathy. The present study was designed to determine whether changes in SGs and Dys might occur in animals with chronic heart failure (CHF) induced by acute myocardial infarction (AMI), which have no genetic defects. METHODS: AMI was induced by the left coronary artery ligation (CAL) in rats. The hemodynamic parameters of the 2- and 8-week CAL (2w- and 8w CAL) rats were measured and the myocardial SGs, Dys, calpain, and calpastatin levels were determined by the Western blot method. Myocardial calpain-like protease activity was evaluated as caseinolysis activity. RESULTS: Increases in left ventricular end-diastolic pressure (LVEDP) and right ventricular systolic pressure, and a decrease in +/-dP/dt were observed at the 2nd week, whereas cardiac output index (COI) was preserved. In contrast, the 8w-CAL rats showed a further increment in LVEDP with low COI. alpha-SG of the viable left ventricle (LV), and septum (Sep) of the 8w-CAL rat decreased (60-70% of the control). The alpha- and beta-SGs of the right ventricle (RV) of the 2w- and 8w-CAL rats were reduced, while gamma- and delta-SGs in the three regions did not change significantly. Dys in the viable LV and RV of the 8w-CAL rat decreased (75% of the control). The amount of m-calpain in the three regions of the 2w- and 8w-CAL rats increased (140-200% of the control), whereas the endogenous calpain inhibitor, calpastatin, did not change significantly. The in vitro degradation studies using purified m-calpain or cytosolic fractions of the 8w-CAL rat heart suggested a reduction in SGs and Dys by calpain. CONCLUSION: The results suggest that a decrease in SGs and Dys may play an important role in the pathophysiology of CHF following AMI. PMID- 12909326 TI - Intracellular ATP is required for mitochondrial apoptotic pathways in isolated hypoxic rat cardiac myocytes. AB - OBJECTIVES: The present study examined the possibility that intracellular ATP levels dictate whether hypoxic cardiac myocytes die by apoptosis or necrosis. BACKGROUND: Although apoptosis and necrosis may appear to be distinct forms of cell death, recent studies suggest that the two may represent different outcomes of a common pathway. In ischemic myocardium, apoptosis appears early, while energy stores are presumably still available, followed only later by necrosis. METHODS: Neonatal rat cardiac myocytes were exposed to continuous hypoxia, during which the intracellular ATP concentration was modulated by varying the glucose content in the medium. The form of cell death was determined at the end of the hypoxic exposure. RESULTS: Under total glucose deprivation, ATP dropped precipitously and cell death occurred exclusively by necrosis as determined by nuclear staining with ethidium homodimer-1 and smearing on DNA agarose gels. However, with increasing glucose concentrations (10, 20, 50, 100 mg/dl) cellular ATP increased correspondingly, and apoptosis progressively replaced necrosis until it became the sole form of cell death, as determined by nuclear morphology, DNA fragmentation on agarose gels, and caspase-3 activation. The data showed a significantly positive correlation between myocyte ATP content and the percentage of apoptotic cells. Hypoxia resulted in lactate production and cellular acidification which stimulates apoptosis. However, acidification-induced apoptosis was also increased in an ATP-dependent fashion. Loss of mitochondrial membrane potential and cytochrome c release from the mitochondria was observed in both the apoptotic and necrotic cells. Furthermore, translocation of Bax from cytosol into mitochondria preceded these events associated with mitochondrial permeability transition. Increased lactate production and a lack of effect by the mitochondrial inhibitor oligomycin indicated that ATP was generated exclusively through glycolysis. CONCLUSIONS: We demonstrate that ATP, generated through glycolysis, is a critical determinant of the form of cell death in hypoxic myocytes, independently of cellular acidification. Our data suggest that necrosis and apoptosis represent different outcomes of the same pathway. In the absence of ATP, necrosis prevails. However, the presence of ATP favors and promotes apoptosis. PMID- 12909327 TI - Effects of sildenafil on myocardial infarct size, microvascular function, and acute ischemic left ventricular dilation. AB - OBJECTIVE: Adverse cardiac events in patients treated with the phosphodiesterase 5 inhibitor sildenafil for erectile dysfunction raised concerns about its safety in ischemic heart disease. METHODS: In anesthetized open-chest rabbits, receiving 1.45 mg/kg sildenafil intravenously or saline 30 min prior to ischemia (n=12, each), infarct size (IS, triphenyltetrazolium), the area of no-reflow (ANR, thioflavin S) (% of the risk area, RA, blue dye), and regional myocardial blood flow (RMBF, radioactive microspheres) were measured after 30 min of coronary occlusion and 180 min of reperfusion. Left ventricular hemodynamics and dimensions (echocardiography) were determined in a separate series of animals (n=5, each). RESULTS: Sildenafil significantly lowered arterial blood pressure before occlusion (-17 to -19 mmHg over 30 min), but during ischemia and reperfusion hemodynamics were comparable to controls. IS in treated animals (51+/ 4%) did not significantly differ from control animals (47+/-4%). No major arrhythmias or lengthening of QT/QTc occurred. While sildenafil slightly increased RMBF and significantly reduced specific vascular resistance in the RA during reperfusion (51+/-7 versus 73+/-10 mmHg g min/ml, P<0.05), the ANR (46+/ 3%) was similar to control animals (44+/-4%). Sildenafil reduced left ventricular dP/dt(max) (P<0.05) and dP/dt(min) (P<0.01) in non-ischemic conditions, and slightly during ischemia, along with a pronounced decrease in ischemic left ventricular end-diastolic pressure (9+/-2 versus 15+/-2 mmHg after saline, P<0.05), but did not attenuate acute ischemic left ventricular dilation. CONCLUSIONS: Sildenafil reduced cardiac pre- and afterload, and parameters of left ventricular contractility. Myocardial necrosis and microvascular dysfunction were neither exacerbated nor attenuated. PMID- 12909328 TI - Brief episode of ischemia activates protective genetic program in rat heart: a gene chip study. AB - OBJECTIVE: Brief episodes of ischemia of 20 min or less have the potential to protect the heart. Such episodes are associated primarily with reversible ischemic injury yet they induce changes in gene expression. The purpose of the study was to determine whether activation of protective genes takes place within 4 h following a brief episode of ischemia that would mimic angina pectoris. METHODS: Three groups of rats were studied. In the control (Ctrl) group, hearts were immediately excised following anesthesia; in the sham-operated (SO) group, opened-chest rats received 4 h and 20 min of no intervention; and in the group subjected to ischemia (SI) hearts received 20 min of proximal coronary occlusion followed by 4 h of reperfusion. Hearts from the SI group were divided into nonischemic (NI) and ischemic (Isc) areas. Changes in gene expression pattern were analyzed by using Affymetrix Gene Chips. RESULTS: Ischemia led to strong upregulation of mRNA transcripts for heat shock proteins 70, 27, 105, 86 and 40 kDa, vascular endothelial growth factor, brain-derived neurotrophic factor, plasminogen activator inhibitor-1, activating transcription factor 3, B-cell translocation gene 2, and growth arrest and DNA damage inducible 45 alpha protein compared to the NI tissue. The majority of mRNAs whose levels increased following brief ischemia were of a protective nature. CONCLUSION: Genetic reprogramming emerging during or following brief episodes of ischemia that simulate angina, can be characterized as protective in nature. Developing new therapeutic strategies aimed to promote this protective response represents a legitimate target for future research. PMID- 12909329 TI - Induction of matrix metalloproteinases-14 and -2 by cyclical mechanical stretch is mediated by tumor necrosis factor-alpha in cultured human umbilical vein endothelial cells. AB - OBJECTIVE: Mechanical forces have profound effects on endothelial cells. This study was undertaken to examine the hypothesis that tumor necrosis factor-alpha (TNF-alpha) is a potential mediator of stretch-induced effects on matrix metalloproteinase (MMP). METHODS: Human umbilical vein endothelial cells (HUVECs) grown on a flexible membrane base were stretched by vacuum to 20% of maximum elongation, at 60 cycles/min. We used the TNF-alpha monoclonal antibody and c-Jun N-terminal kinase (JNK) inhibitor, SP600125, to investigate the cyclical stretch induced expression of MMP-14 and -2 in cultured HUVECs. RESULTS: Cyclical mechanical stretch significantly increased protein synthesis and mRNA expression for MMP-14 and -2 from 2 to 24 h. The increased MMP-14 and-2 proteins after stretch were completely blocked after the addition of TNF-alpha monoclonal antibody (5 microg/ml) or SP600125 (20 microM) 30 min before stretch. By zymography, MMP-2 expression was induced by cyclical stretch and was attenuated by TNF-alpha monoclonal antibody and SP600125. Cyclical stretch increased the immunohistochemical labeling of MMP-14 and -2 and significantly increased release of TNF-alpha into the culture media from 120+/-2 to 331+/-2 pg/ml (P<0.001) after stretch for 12 h. Cyclical stretch increased and SP600125 decreased the phosphorylated JNK. Gel-shifting assay showed that DNA-protein binding activity of AP-1 increased after cyclical stretch and TNF-alpha monoclonal antibody and SP600125 abolished the binding activity induced by cyclical stretch. CONCLUSION: These findings indicate that cyclical stretch augments TNF-alpha production and MMP genes expression in HUVECs. TNF-alpha mediates the stretch-induced MMP genes expression, at least in part, through the JNK pathway. PMID- 12909330 TI - ATP antagonism of thrombin-induced endothelial barrier permeability. AB - OBJECTIVES: Thrombin induces endothelial barrier failure by activating the contractile machinery of endothelial cells. Contractile activation is due to an increase in myosin light chain (MLC) phosphorylation. Here, it was investigated whether stimulation of endothelial cells with ATP can interrupt this thrombin induced pathomechanism. METHODS: In cultured human umbilical vein endothelial cells, cytosolic calcium [Ca(2+)](i) (Fura 2 method), phosphorylation of MLC, isometric tension and permeability for albumin were studied. RESULTS: Thrombin (0.2 U/ml) increased [Ca(2+)](i) from a basal level of 78+/-8 to 570+/-63 nM (mean+/-S.D., n=5, P<0.05), MLC phosphorylation from 71+/-7 to 163+/-18%, isometric tension from 157+/-17 to 232+/-26 microN, and permeability from 2.8+/ 0.4 to 11.6+/-1 x 10(-6) cm/s. Co-presence of ATP (10 microM) and thrombin did not alter the [Ca(2+)](i) rise, but reduced MLC phosphorylation to 59.8+/-10%, isometric tension to 174+/-14 microN, and permeability to 5.4+/-0.6 x 10(-6) cm/s. The thrombin-induced rise in MLC phosphorylation was sensitive to reduction of [Ca(2+)](i) It was accompanied by an increase in Rho activation, and was inhibited by Y-27632 (10 microM), a Rho-kinase blocker. The ATP-induced decrease in MLC phosphorylation was not sensitive to [Ca(2+)](i). It was not accompanied by changes in RhoA activation, and could not by suppressed by Y-27632. CONCLUSIONS: ATP antagonizes the Ca(2+)- and Rho-dependent effects of thrombin on MLC phosphorylation most likely by a Ca(2+)- and Rho-independent activation of MLC phosphatase. It thereby functionally antagonizes the thrombin-induced increase in monolayer tension and permeability. PMID- 12909331 TI - Involvement of cyclin-dependent pathway in the inhibitory effect of delphinidin on angiogenesis. AB - OBJECTIVE: Epidemiologic studies have shown that a diet rich in fruits and vegetables has a beneficial preventive effect for cancer and cardiovascular diseases. The mechanisms of these beneficial effects are not known although there is evidence that polyphenolic compounds in food may be of some benefit. The purpose of this study was to define the effect of delphinidin, a vasoactive polyphenol belonging to the class of anthocyanin, on endothelial cell proliferation and migration as well as on in vivo angiogenesis. METHODS AND RESULTS: Vascular endothelial growth factor-stimulated human umbilical endothelial cell migration and proliferation are potently inhibited by delphinidin. Flow cytometric analysis demonstrates that delphinidin inhibition of proliferation is correlated with the blockade of cell cycle in G(0)/G(1) phase. Western blot analysis shows that delphinidin reverses the vascular endothelial growth factor-induced decrease in expression of cyclin-dependent kinase inhibitor p27(kip1) and the vascular endothelial growth factor-induced increase of cyclin D1 and cyclin A, both being necessary to achieve the G(1)-to-S transition. Furthermore, delphinidin inhibits neovascularisation in vivo in chorioallantoic membrane model. CONCLUSION: Delphinidin overcomes in vitro and in vivo angiogenesis and thus appears promising for the development of an anti-angiogenic therapy. PMID- 12909332 TI - Impaired activities of antioxidant enzymes elicit endothelial dysfunction in spontaneous hypertensive rats despite enhanced vascular nitric oxide generation. AB - OBJECTIVE: Enhanced oxidative stress is involved in mediating the endothelial dysfunction associated with hypertension. The aim of this study was to investigate the relative contributions of pro-oxidant and anti-oxidant enzymes to the pathogenesis of endothelial dysfunction in genetic hypertension. METHODS: Dilator responses to endothelium-dependent and endothelium-independent agents such as acetylcholine (ACh) and sodium nitroprusside were measured in the thoracic aortas of 28-week-old spontaneously hypertensive rats (SHR) and their matched normotensive counterparts, Wistar Kyoto rats (WKY). The activity and expression (mRNA and protein levels) of endothelial nitric oxide synthase (eNOS), p22-phox, a membrane-bound component of NAD(P)H oxidase, and antioxidant enzymes, namely, superoxide dismutases (CuZn- and Mn-SOD), catalase and glutathione peroxidase (GPx), were also investigated in aortic rings. RESULTS: Relaxant responses to ACh were attenuated in phenylephrine-precontracted SHR aortic rings, despite a 2-fold increase in eNOS expression and activity. Although the activity and/or expression of SODs, NAD(P)H oxidase (p22-phox) and GPx were elevated in SHR aorta, catalase activity and expression remained unchanged compared to WKY. Pretreatment of SHR aortic rings with the inhibitor of xanthine oxidase, allopurinol, and the inhibitor of cyclooxygenase, indomethacin, significantly potentiated ACh-induced relaxation. Pretreatment of SHR rings with catalase and Tiron, a superoxide anion (O(2)(-)) scavenger, increased the relaxant responses to the levels observed in WKY rings whereas pyrogallol, a O(2)(-)-generator, abolished relaxant responses to ACh. CONCLUSION: These data demonstrate that dysregulation of several enzymes, resulting in oxidative stress, contributes to the pathogenesis of endothelial dysfunction in SHR and indicate that the antioxidant enzyme catalase is of particular importance in the reversal of this defect. PMID- 12909333 TI - Tetrahydrobiopterin and antioxidants reverse the coronary endothelial dysfunction associated with left ventricular hypertrophy in a porcine model. AB - OBJECTIVE: Endothelium-dependent G-protein mediated relaxations of epicardial coronary arteries is impaired with left ventricular hypertrophy. The objective of this study was to assess the effect of L-arginine, BH(4) and the combination of two antioxidants, superoxide dismutase and catalase, on endothelium-dependent relaxations in a swine left ventricular hypertrophy model. METHODS: Aortic banding was performed 3 cm above the coronary ostia. Vascular reactivity studies were performed in standard organ chamber experiments to assess the NO pathway in the presence of methyltetrahydropterin (a BH(4) analogue), L-arginine, superoxide dismutase and catalase. RESULTS: There was a statistically significant increase in endothelium-dependent relaxation to serotonin and to bradykinin with methyltetrahydropterin and with superoxide dismutase plus catalase (P<0.05) but not with L-arginine compared to untreated coronary arteries from left ventricular hypertrophy animals. Plasma 3-nitrotyrosine level increased significantly from 918+/-122 to 1844+/-300 microM (P<0.05 vs. control) after 60 days of aortic banding. Endothelial dysfunction was not associated with a reduced expression of endothelial nitric oxide synthase 2 months after pressure overload left ventricular hypertrophy. CONCLUSIONS: Treatment with BH(4) and antioxidants constitutes an interesting approach for the prevention of endothelial dysfunction in epicardial coronary arteries associated with left ventricular hypertrophy. PMID- 12909334 TI - Mechanism of E2F1-induced apoptosis in primary vascular smooth muscle cells. AB - OBJECTIVE: The transcription factor E2F1 serves as a major regulator of the cell cycle by controlling G1-S phase transition. However, apart from its proliferative function high levels of deregulated E2F1 are capable of inducing apoptosis depending on the cellular context. In particular the tumor suppressor p53 and its homologue p73 are implicated in this proapoptotic function. METHODS: Here, we investigated the mechanistic basis for E2F1-mediated apoptosis in vascular smooth muscle cells (VSMCs) which have previously been shown to be E2F1-responsive. RESULTS: Interestingly, E2F1-expression in these cells induced clear signs of apoptosis in the absence of any proliferative activity. Although cell-cycle regulated genes such as CCNE1 and CDC25A were activated, BrdU-staining revealed no S-phase entry. Instead, a rapid loss of cell viability by induction of apoptosis was observed. Using a transactivation-defective E2F1-mutant, we show that apoptosis induction is independent of the transactivation function and therefore independent of ARF and p73. However, this mutant retains its ability to stabilize and phosphorylate p53, suggesting that p53 is sufficient for the effect of E2F1. CONCLUSION: VSMCs therefore represent a cellular system in which the transactivation-independent, proapoptotic activity of E2F1 is the primary cellular function. Ectopic expression of E2F1 might therefore be a suitable therapy to prevent VSMC hyperproliferation. PMID- 12909335 TI - Influence of differential vascular remodeling on the coronary vasomotor response. AB - OBJECTIVES: Arterial remodeling may increase or decrease the luminal encroachment of atherosclerotic plaques in the coronary circulation. However, the factors determining the nature and consequences of the remodeling process remain poorly characterized. The study aims were to assess whether the pattern of vascular remodeling influences the physical and vasomotor responses of the coronary arteries in vivo in man. METHODS: Coronary vessel area, distensibility and stiffness were determined in positively, negatively and non-remodeled arterial segments using intravascular ultrasound and Doppler flow measurement. Epicardial vasomotor responses were determined following intracoronary boluses of acetylcholine (10(-6) and 10(-4) M), adenosine (24-30 microg) and nitroglycerin (200 microg). RESULTS: Fifty-six coronary arterial segments were studied in 25 patients. In comparison to non- and positively remodeled segments, negatively remodeled segments had a higher stiffness index (67+/-16 vs. 33+/-5 and 38+/-8, respectively; P<0.02) and appeared to have lower compliance and distensibility (0.66+/-0.17 vs. 1.65+/-0.54 and 0.94+/-0.18/mmHg; P=NS). Non-remodeled segments had a greater change in vessel area with 10(-6) M acetylcholine (4.9+/-0.8%), compared to positively and negatively remodeled segments (0.6+/-1.8% and -4.9+/ 1.8%, respectively, P<0.05). A significant degree of preservation of vasodilatation to 10(-6) M acetylcholine was evident in positively remodeled compared with negatively remodeled segments (P<0.05). Nitroglycerin caused greater vasodilatation in non-remodeled segments (7.2+/-3.8%) than either positively or negatively remodeled segments (4.7+/-0.9 and 3.7+/-0.6%, respectively, P<0.05). CONCLUSIONS: Vascular remodeling is an important and major determinant of local epicardial vasomotor responses. Both structural and functional abnormalities are associated with negative remodeling that may contribute to the adverse effects of such lesions. PMID- 12909336 TI - DExD/H-box proteins and their partners: helping RNA helicases unwind. AB - Members of the DExD/H-box family of RNA helicases are involved in many processes and complexes within the cell. While individual DExD/H helicase family members have been studied extensively, the mechanisms through which helicases affect multiprotein complexes are just beginning to be investigated. Because RNA helicases are both highly conserved and numerous in the cell, study of RNA helicase recruitment and modulation by cofactors is necessary for understanding the mechanisms of helicase action in vivo. This review will focus on cofactor mediated regulation of helicase target specificity and activity. PMID- 12909337 TI - Regulation of gene expression by stop codon recoding: selenocysteine. AB - The regulation of gene expression at the translational level not only allows for rapid changes in specific protein levels but also provides an opportunity to alter codon specificity. For the incorporation of selenocysteine (Sec) into protein, the UGA codon is transformed from one that signals translation termination to one specific for Sec. This review provides a look at Sec incorporation from the perspective of the individual steps involved in protein synthesis: initiation, elongation and termination. The roles of the factors known to be required for Sec incorporation are considered in the context of each step in translation including structural modeling of the differences between the standard elongation factor eEF1A and the Sec-specific counterpart, eEFSec. PMID- 12909338 TI - A HANDful of questions: the molecular biology of the heart and neural crest derivatives (HAND)-subclass of basic helix-loop-helix transcription factors. AB - The HAND subclass of basic Helix-loop-helix factors is comprised of two members HAND1 and HAND2. HAND genes are present within the genomes of organisms ranging from flies to man. Experiments employing chick embryology, tissue culture, and gene targeting in mice show that HAND function is critical for the specification and/or differentiation of extraembryonic structures that include the yolk sac, placenta, and the cells of the trophoblast lineages. HAND factors also play key roles in cardiac, gut, sympathetic neuronal development and in the proper development of tissues populated by HAND-expressing neural crest cells, including regions of the developing vasculature, the limbs, the jaw, and teeth. Surprisingly, nearly 10 years after their initial identification and characterization, little is understood about the nature of the downstream target genes which HAND1 and HAND2 regulate, whether the nature of their transcriptional regulation is positive or negative, or if they modulate genetic programs common to these diverse tissue types or if they drive unique subsets of genes that contribute to tissue identity. At the core of these questions is by which mechanisms do HAND factors modulate biological activity? Do they behave like classical class B bHLH factors or is their function more complex requiring a rethinking of the dogma? What follows is a review of what is currently known about HAND factors and a reflection on why elucidating their role in the biological programs within which they participate has been so difficult. PMID- 12909339 TI - Juxtacentromeric region of human chromosome 21: a boundary between centromeric heterochromatin and euchromatic chromosome arms. AB - We have analysed the genomic structure and transcriptional activity of a 2.3-Mb genomic sequence in the juxtacentromeric region of human chromosome 21. Our work shows that this region comprises two different chromosome domains. The 1.5-Mb proximal domain: (i) is a patchwork of chromosome duplications; (ii) shares sequence similarity with several chromosomes; (iii) contains several gene fragments (truncated genes having an intron/exon structure) intermingled with retrotransposed pseudogenes; and (iv) harbours two genes (TPTE and BAGE2) that belong to gene families and have a cancer and/or testis expression profile. The TPTE gene family was generated before the branching of Old World monkeys from the great ape lineage, by intra- and interchromosome duplications of the ancestral TPTE gene mapping to phylogenetic chromosome XIII. By contrast, the 0.8-Mb distal domain: (i) is devoid of chromosome duplications; (ii) has a chromosome 21 specific sequence; (iii) contains no gene fragments and only one retrotransposed pseudogene; and (iv) harbours six genes including housekeeping genes. G-rich sequences commonly associated with duplication termini cluster at the boundary between the two chromosome domains. These structural and transcriptional features lead us to suggest that the proximal domain has heterochromatic properties, whereas the distal domain has euchromatic properties. PMID- 12909340 TI - The 200-kb segmental duplication on human chromosome 21 originates from a pericentromeric dissemination involving human chromosomes 2, 18 and 13. AB - Regions close to human centromeres contain DNA fragments spanning hundreds of kilobases that exhibit a high degree of sequence identity (>95%). Here we report the genomic structure and evolution of a family of four paralogous regions related to a 220-kb genomic fragment present on the long arm of human chromosome 21 (21q22.1). Phylogenetic classification of the paralogous sequences obtained from the draft of the Human Genome Project are in agreement with results from comparative fluorescence in situ hybridization on metaphase chromosomes from human and great apes. The original copy present in 21q22.1 in human was duplicated in great apes after the divergence of the orang-utan and inserted in a pericentromeric region, most likely the ancestor of HSA2q, then disseminated by transposition of a larger fragment to other pericentromeric locations: HSA18p11, HSA13q11 and HSA21q11.1. The degree of dissemination varies among species. PMID- 12909341 TI - The human genome has 49 cytochrome c pseudogenes, including a relic of a primordial gene that still functions in mouse. AB - Using a computational approach, we have identified 49 cytochrome c (cyc) pseudogenes in the human genome. Analysis of these provides a detailed description of the molecular evolution of the cyc gene. Almost all of the pseudogenes are full-length, and we have concluded that they mostly originated from independent retrotransposition events (i.e. they are processed). Based on phylogenetic analysis and detailed sequence comparison, we have further divided these pseudogenes into two groups. The first, consisting of four young pseudogenes that were dated to be between 27 and 34 Myr old, originated from a gene almost identical to the modern human cyc gene. The second group of pseudogenes is much older and appears to have descended from ancient genes similar to modern rodent cyc genes. Thus, our results support the observation that accelerated evolution in cyc sequence had occurred in the primate lineage. The oldest pseudogene in the second group, dated to be over 80 Myr old, resembles the testis-specific cyc gene in modern rodents. It is likely that the mammalian ancestor had both the somatic and the testis-specific cyc genes. While the testis specific gene is still functional in modern rodents, the human has lost it, retaining only a pseudogene in its place. Thus, our study may have identified a pseudogene that is a dead relic of a gene that has completely died off in the human lineage. PMID- 12909342 TI - Characterisation of a gene cluster in Fugu rubripes containing the complement component C4 gene. AB - In this study, we describe the characterisation of the complement component C4 gene in Fugu rubripes. The Fugu C4 gene, orthologous to the tetrapod C4 gene, encompasses a genomic span of 9702 base pairs and contains 41 exons, encoding the typical C4 three-chain polypeptide. The gene encodes a protein containing 1703 amino acids. The Fugu C4 protein demonstrates the presence of 25 conserved cysteine residues, as well as conservation of the functionally important thioester site. Complete sequencing of one cosmid and sequence scans from a cluster of 18 overlapping BAC clones, centering around the C4 gene, have identified the short-range linkage with five orthologous human genes mapping to the Major Histocompatibility Complex (MHC) including: tenascin X (TNX); cytochrome P450, subfamily XXIA, polypeptide 2 (CYP21A2); allograft inflammatory factor 1 (AIF1) and casein kinase 2, beta polypeptide (CSNK2B), all found in the MHC class III region; and retinoid X receptor, beta (RXRB),which resides in the MHC extended class II region. To date, this syntenic association of the Fugu C4 and other MHC class III region genes has not been observed in other teleost fish. Data from the recent whole-genome shotgun assemblies reveal the Fugu MHC-related cluster of genes to be flanked predominantly by genes mapping to human chromosomes 7 and 19. All of the six identified Fugu MHC-related genes have been characterised at the genomic level. PMID- 12909343 TI - Evolution of low-copy number and major satellite DNA sequences coexisting in two Pimelia species-groups (Coleoptera). AB - Satellite DNA sequence evolution has been studied in several insect species from the genus Pimelia (Tenebrionidae, Coleoptera). Low-copy number homologs of the previously characterized major satellite DNA from P. monticola (PMON) have been cloned and sequenced from six congeneric species belonging to two species groups: Ibero-Balearic and Moroccan. Sequence analysis of a sample of low-copy number repeats revealed two subfamilies, differing on average 17.5% due to randomly spread single point mutations. Each subfamily is specific for a group of taxa in congruence with their biogeography. Within each group, there is no significant species-specific clustering of the sequences. These results suggest that the two satellite subfamilies arose after the split of an ancestral lineage into the North African and Ibero-Balearic Pimelia species-groups, but before their subsequent radiation. Rate heterogeneity tests suggest that PMON sequences have evolved faster in the lineage leading to the Moroccan group. Comparison of sequence divergences between minor PMON and the previously characterized major PIM357 satellite obtained from the same taxa, points to similar evolutionary dynamics. Both sequences are evolving in parallel accumulating mutations in a gradual manner irrespectively of significant differences in abundance. These data show that copy number of the sequence families does not necessarily affect the sequence change dynamics of satellite repeats. PMID- 12909344 TI - A third isoform of cytochrome c oxidase subunit VIII is present in mammals. AB - The terminal enzyme of the mitochondrial respiratory chain, cytochrome c oxidase (COX), contains three mitochondrial and ten nuclear encoded subunits in mammals. Three of the nuclear subunits (VIa, VIIa, and VIII) have muscle and non-muscle specific isoforms, subunit IV contains a lung-specific isoform, and subunit VIb contains a testes-specific isoform. For subunit VIII, the smallest nuclear encoded COX polypeptide, we have now found a third gene (COX 8-3), which has been identified in human, lemur, rat, and mouse, suggesting that it is present in a broad range of Eutherian mammals. Sequence similarity and gene structure support the homology of COX8-3 to the other subunit VIII isoforms, indicating that all three are the product of gene duplications. COX VIII-3 protein is mitochondrially targeted, as shown by a fluorescent COX VIII3/DsRed fusion protein. Both the mitochondrial targeting and its sequence conservation suggest that COXVIII-3 functions as part of the COX holoenzyme and could have a tissue-specific role, as is the case for the other two isoforms. Questions remain about where COX8-3 is predominantly expressed. However, detection of full-length cDNAs, lower levels of sequence divergence at the first and second codon positions compared to the third, and a conserved gene structure indicate that COX VIII-3 is an expressed gene whose origin dates to at least 91 million years ago. PMID- 12909345 TI - Cloning and characterization of rice HMGB1 gene. AB - We isolated a 918 bp long full-length rice HMGB1 cDNA, which has an open reading frame of 471 bp encoding 157 amino acids, with a central domain of high sequence similarity to the HMG-box domain of other plant HMGB1 proteins. RNA gel blot analysis indicated that rice HMGB1 gene is constitutively expressed in various tissues and organs. Southern hybridization and sequence analyses suggested that a single copy of the HMGB1 gene composed of seven exons and six introns exists in rice. We have also cloned a 1755 bp long 5' flanking region of the rice HMGB1 gene, which can be regarded as its promoter. 5' deletion analysis of this promoter indicated that positive cis-elements residing between -1400 and -1115 are important to enhance quantitative expression, whereas negative cis-elements between -1755 and -1400 and between -1115 and -351 inhibit expression. PMID- 12909346 TI - An ascidian RING finger gene is specifically expressed in a single cell of larval ocellus. AB - The ascidian nervous system is extremely simple, although the structure of it is comparable with the complex vertebrate nervous system. This simplicity makes the ascidian nervous system a good model to understand how the neuronal circuit is built up in the chordate nervous system. In order to study the formation of the neuronal circuit at the single cell level, molecular markers to characterize specific single cells are desired. In the present paper, we describe the gene expression pattern of CIGL: an ascidian homologue of Goliath, a Drosophila RING finger gene. In the early embryonic stage, CiGl is expressed in the lateral part of the neural tube and in several peripheral nerve cells. Later in the larval stage, CiGl specifically marks ocellus: one of the pigment cells in the ascidian brain, which is involved in the photoreceptive system. CiGl will be useful to understand the differentiation mechanism of ocellus, and especially to test the model proposed by. In addition, the finding of this single cells specific gene expression pattern at a certain developmental stage encourages us to look for more genes which mark single cells, especially those that have not been well characterized. PMID- 12909347 TI - Cloning and characterization of a novel member of the human ATF/CREB family: ATF2 deletion, a potential regulator of the human DNA polymerase beta promoter. AB - The solitary cAMP response element (CRE)1 in the human DNA polymerase beta (beta pol) core promoter plays a key role in both basal expression and the DNA alkylating agent response of the promoter. To further understand the role of the CRE in the regulation of this promoter, we searched for novel CRE-binding proteins by using a 32P-labeled beta-pol CRE oligodeoxynucleotide and a human cDNA expression library constructed in phage lambda. A total of fourteen phage clones were isolated, corresponding to various members of the CRE-binding protein family. One of these clones, termed ATF2 deletion (ATF2d), encodes a novel ATF2 isoform and was chosen for further characterization in this study. Relative to ATF2 mRNA, this clone contains an internal 97-nt deletion and a unique 3' region. The 97-nt deletion causes a frame shift, resulting in a ATF2-like polypeptide of approximately 60 kDa. ATF2d retains the bZIP domain of ATF2, lacks the N-terminal zinc-finger region, and includes novel characteristics in its N- and C-terminal regions. PMID- 12909348 TI - Characterization of mouse Eppin and a gene cluster of similar protease inhibitors on mouse chromosome 2. AB - We have recently described a novel gene on human chromosome 20q 12-13.2 called Eppin (Epididymal protease inhibitor) that expresses three mRNAs encoding two isoforms of a cysteine-rich protein containing both Kunitz-type and WAP-type (four disulfide core) consensus sequences (Richardson et al., 2001). To further our studies on Eppin, we have cloned, sequenced and characterized mouse Eppin and report that it lies within a 200 Kb cluster of putative Eppin-like genes on mouse chromosome 2. Analysis of the homologies between the genes in the human and mouse Eppin clusters indicates that the first part of the cluster immediately surrounding Eppin represents a conserved linkage because the order of homologous genes is conserved. Sequencing of reverse transcription polymerase chain reaction (RT-PCR) products confirmed the expression of five of these novel Eppin-like genes in the mouse, which include the mouse homologue of HE-4. These genes are characterized by having either one or both of the Kunitz-type and WAP-type consensus sequences. Additional RT-PCR experiments revealed that expression of some of the Eppin-like genes is restricted to epididymis and testis while others are expressed in several somatic tissues. Northern blot analysis of 22 different mouse tissues identified Eppin transcripts only in the epididymis and testis. Immunostaining of Eppin with anti-recombinant mouse Eppin demonstrated Eppin predominantly on the postacrosomal region of mouse spermatozoa, in Sertoli cells, Leydig cells, and round spermatids in the testis, and in the principal cells of the cauda epididymidis epithelium. Eppin is first expressed by Sertoli cells of 12-day-old mice and subsequently in round spermatids, which is consistent with androgen regulation. Our results demonstrate that mouse chromosome 2 contains a conserved linkage of Eppin-like protease inhibitor genes that are expressed in the epididymis. PMID- 12909349 TI - Differential expression of human replacement and cell cycle dependent H3 histone genes. AB - Histones are the major protein component of chromatin. Except H4, all histone classes consist of several subtypes. The H3 family includes two replacement histone genes, H3.3A and H3.3B, which both encode the same protein and are expressed independently from the cell cycle. Since the two genes encode an identical protein, we analyzed whether they are differentially expressed. Therefore we cloned, sequenced and characterized the regulatory structures of the H3.3A gene and compared these with the corresponding regions in the H3.3B gene. In contrast to the H3.3B promoter, the promoter region of the H3.3A gene revealed neither a TATA nor any CCAAT boxes but an initiator element and several SP1 binding sequence motifs within an overall GC-rich sequence. Northern blot analysis of RNA from six human cell lines revealed that every cell line expressed each of the H3 isoform genes H3.1, H3.3A and H3.3B. In contrast, analysis of total RNA from human tissues showed a differential expression of the H3 isoform genes. The H3.3 genes are essentially only expressed in adult tissue, whereas the H3.1 gene is transcribed just in fetal tissue. The functional relevance of the elements identified by sequence analysis was established using a reporter gene assay with deletion constructs of the H3.3A promoter. In this assay a 256 bp fragment was sufficient for the full promoter activity and three promoter segments, each containing SP1 binding motifs, contribute to the H3.3A gene expression. The possible functional relevance of the differences between the two H3.3 genes in structure and expression is discussed. PMID- 12909350 TI - Expression of a novel type I keratin, DAPK-1 in the dorsal aorta and pronephric duct of the zebrafish embryos. AB - We isolated a novel cytokeratin gene of zebrafish (Danio rerio), DAPK-1 closely related to other vertebrate type I cytokeratin genes. Zygotic transcription starts at the sphere stage. After the mid-blastula stage, DAPK-1 is expressed in all surface cells, notably in those of the outer enveloping layer. DAPK-1 messages are also present specifically during the segmentation, pharyngula, and hatching periods. In particular, after 24 h post-fertilization, its expression is restricted to the developing eye region, otic vesicle, pectoral fin, dorsal aorta, and pronephric duct. In the mindbomb mutant embryo that has defects in the dorsal aorta development, DAPK-1 transcripts are not detected in the dorsal aorta and pronephric duct. The characteristic expression pattern of DAPK-1 may facilitate more detailed studies related to the morphogenesis of dorsal aorta and pronephric duct. PMID- 12909351 TI - Bacterial type III secretion systems are ancient and evolved by multiple horizontal-transfer events. AB - Type III secretion systems (TTSS) are unique bacterial mechanisms that mediate elaborate interactions with their hosts. The fact that several of the TTSS proteins are closely related to flagellar export proteins has led to the suggestion that TTSS had evolved from flagella. Here we reconstruct the evolutionary history of four conserved type III secretion proteins and their phylogenetic relationships with flagellar paralogs. Our analysis indicates that the TTSS and the flagellar export mechanism share a common ancestor, but have evolved independently from one another. The suggestion that TTSS genes have evolved from genes encoding flagellar proteins is effectively refuted. A comparison of the species tree, as deduced from 16S rDNA sequences, to the protein phylogenetic trees has led to the identification of several major lateral transfer events involving clusters of TTSS genes. It is hypothesized that horizontal gene transfer has occurred much earlier and more frequently than previously inferred for TTSS genes and is, consequently, a major force shaping the evolution of species that harbor type III secretion systems. PMID- 12909352 TI - A novel myosin heavy chain gene in human chromosome 19q13.3. AB - A human myosin heavy chain gene was identified in chromosome 19q13 by computational sequence analysis, RT-PCR and DNA sequencing of the cDNA. The complete cDNA has a length of 6786 bp and comprises 41 exons (40 coding) included in 108 kb of genomic sequence. Alternative splicing variants were also identified. The gene is expressed in a multitude of tissues, but mainly in small intestine, colon and skeletal muscle. The putative protein (228 kDa) carries the common myosin domains and presents high homology with the non-muscle myosin heavy chains (MYH9 and MYH10) as well as the smooth muscle myosin heavy chain MYH11. Nevertheless, phylogenetic analysis indicated that these homologous proteins are more related among themselves than to MYH14, suggesting that possibly this myosin heavy chain should be classified in a new myosin-subfamily. PMID- 12909353 TI - Pneumocystis carinii STE11, an HMG-box protein, is phosphorylated by the mitogen activated protein kinase PCM. AB - A pheromone-induced mitogen activated protein kinase (MAPK) pathway controls mating in fungi by regulating gene transcription. In the opportunistic fungus Pneumocystis carinii, we have identified a protein containing a high-mobility group (HMG) motif which is homologous to the transcriptional activators STE11 of Schizosaccharomyces pombe and STE12 of Saccharomyces cerevisiae. In fungi, this transcriptional activator functions in sexual development, filamentous growth, and pathogenicity. The fungal pheromone-activated MAPK phosphorylates the transcriptional activator to allow binding to pheromone-response elements in the promoter regions of certain genes. We have previously identified a P. carinii MAPK, PCM, which has significant homology to fungal MAPKs involved in mating. As an initial step in understanding the downstream molecules which interact with the PCM kinase, we have cloned a STE11 homologue in P. carinii. PCSTE11 has an open reading frame of 1.5 kb which encodes a protein of 501 amino acids with a molecular weight of 56 kDa. Greatest homology was to S. pombe STE11 (52%). We have expressed a His-tag fusion of PCSTE11 and purified the protein with nickel affinity resin. PCM phosphorylates the purified protein indicating that PCSTE11 is associated with the MAPK cascade in P. carinii. PMID- 12909354 TI - Cloning and characterization of a novel lipase from Vibrio harveyi strain AP6. AB - A lipolytic enzyme designated Vst, was cloned from Vibrio harveyi strain AP6. The vst gene was 1650 bp and was predicted to encode a 549 amino acid precursor with a molecular mass of about 61 kDa. The predicted polypeptide shared about 30% sequence identity with Bacillus esterases. Sequence alignment of Vst and related esterases revealed the presence of an active site serine located in the middle of the GESAG motif, at positions 212-216. This motif resembled the GXSXG consensus motif characteristic of lipolytic enzymes. Vst was expressed as a carboxy terminal 6 x His tagged recombinant enzyme from E. coli TOP10 cells. SDS-PAGE and Western blot analysis using anti-His antibodies confirmed that the size of the mature protein was about 61 kDa. Substrate specificity of Vst was investigated using p -nitrophenyl (p NP) esters with varying carbon chain lengths, from C2 to C18. Vst showed the highest activity with the long chain p -nitrophenyl ester p NPC12 but was able to hydrolyze longer chain esters (p NPC14-p NPC16) as well as short and medium chain esters (p NPC4 and p NPC8). PMID- 12909355 TI - The 62-kb upstream region of Bombyx mori fibroin heavy chain gene is clustered of repetitive elements and candidate matrix association regions. AB - We sequenced an 80 kb DNA region containing the complete sequence of the silkworm Bombyx mori fibroin gene and its flanking, especially the upstream, regions (-62 kb). About 30% of the 62 kb upstream region is composed of repetitive elements including short interspersed elements Bm1, long interspersed elements L1Bm and mariner-like elements Bmmar1 which are widespread over the silkworm genome. This 62 kb region is also enriched of commonly considered matrix association region (MAR) motifs. A total of 25 individual MAR recognition signatures (MRSs) were identified, with 24 at the upstream and one at the downstream region. Combining two newly developed MAR prediction programs (MAR-finder and Chrclass), ten candidate MARs were predicted, with five containing MRS and seven related to the repetitive elements. The wide distribution of nested repetitive elements, candidate MARs, DNase I hypersensitive sites and other potential regulatory factors recognition sites indicates this region is probably a unique huge cis acting element contributing to the regulation of the spatial and temporal specificity and efficiency of fibroin gene expression. PMID- 12909357 TI - Investigating single nucleotide polymorphism (SNP) density in the human genome and its implications for molecular evolution. AB - We investigated the single nucleotide polymorphism (SNP) density across the human genome and in different genic categories using two SNP databases: Celera's CgsSNP, which includes SNPs identified by comparing genomic sequences, and Celera's RefSNP, which includes SNPs from a variety of sources and is biased toward disease-associated genes. Based on CgsSNP, the average numbers of SNPs per 10 kb was 8.33, 8.44, and 8.09 in the human genome, in intergenic regions, and in genic regions, respectively. In genic regions, the SNP density in intronic, exonic and adjoining untranslated regions was 8.21, 5.28, and 7.51 SNPs per 10 kb, respectively. The pattern of SNP density based on RefSNP was different from that based on CgsSNP, emphasizing its utility for genotype-phenotype association studies but not for most population genetic studies. The number of SNPs per chromosome was correlated with chromosome length, but the density of SNPs estimated by CgsSNP was not significantly correlated with the GC content of the chromosome. Based on CgsSNP, the ratio of nonsense to missense mutations (0.027), the ratio of missense to silent mutations (1.15), and the ratio of non-synonymous to synonymous mutations (1.18) was less than half of that expected in a human protein coding sequence under the neutral mutation theory, reflecting a role for natural selection, especially purifying selection. PMID- 12909356 TI - Low hanging fruit: a subset of human cSNPs is both highly non-uniform and predictable. AB - We present a point mutation classification method that contrasts SNP databases and has the potential to illuminate the relative mutational load of genes caused by codon bias. We group point variation gleaned from public databases by their wild-type and mutant codons, e.g. codon mutation classes (CMCs, 576 possible such as ACG-->ATG), whose frequencies in a database are assembled into a BLOSUM-style matrix describing the likelihood of observing all possible single base codon changes as tuned by the intertwined effects of mutation rate and selection. The rankings of the CMCs in any database are reshuffled according to the population stratification of the typical genotyping experiment producing that resource's data. Analysis of four independent databases reveals that a considerable fraction of mutation in functional genes can be described by a few CMCs regardless of gene identity or population stratification in the genotyping experiment. For example, the top 5% (29/576) of CMCs account for 27.4% of the observed variants in dbSNP while the bottom 5% account for only 0.02%. For non-synonymous disease-causing mutation, 40.8% are described by the top 5% of all possible non-silent CMCs (22/438). Overall, the most observed polymorphism is a G-->A transition at CpG dinucleotides causing ACG, TCG, GCG, and CCG to frequently undergo silent mutation in any gene due to the putative lack of impact on the protein product. In order to assess how well CMC spectrums estimate the aggregate non-synonymous mutational trends of a single gene, a CMC matrix was applied to seven unrelated genes to compute the most likely point mutations. In excess of 87% of these mutation predictions are historically known to play an important role in a disease state according to published literature. CMC-based mutation prediction may aid design and execution of direct association genotyping studies. PMID- 12909358 TI - Identification and cloning of genes encoding viomycin biosynthesis from Streptomyces vinaceus and evidence for involvement of a rare oxygenase. AB - The tuberactinomycins are a family of basic cyclic peptides that exhibit potent antitubercular activity. These peptides are characterized by the presence of an amino acid with a 6-membered cyclic guanidine side chain (capreomycidine) and two or more 2,3-diaminopropionate residues. Viomycin (tuberactinomycin B) is a well studied member of the family, was once prescribed for the treatment of tuberculosis, and has been shown to block translocation during protein biosynthesis. The gene cluster encoding viomycin biosynthesis was identified and cloned from Streptomyces vinaceus. The cluster was identified by screening genomic libraries with the viomycin phosphotransferase self-resistance gene (vph) and non-ribosomal peptide synthetase (NRPS) gene probes amplified from S. vinaceus genomic DNA. The viomycin cluster was localized to ca. 120 kb of contiguous DNA defined by four overlapping cosmid inserts. Each cosmid hybridized with one or more peptide synthetase gene probes and two also hybridized with vph. Confirmation that the cluster encoded viomycin biosynthesis was obtained from the disruption of two NRPS adenylation domains. Partial sequence analysis revealed an ORF (svox) predicted to encode a rare non-heme iron, alpha-ketoglutarate dependent oxygenase proposed to function in the oxidative cyclization of arginine to the capreomycidine residue. Insertional disruption of svox resulted in complete loss of viomycin production, confirming its involvement in the pathway. PMID- 12909360 TI - Molecular characterization of an inducible gentisate 1,2-dioxygenase gene, xlnE, from Pseudomonas alcaligenes NCIMB 9867. AB - Pseudomonas alcaligenes NCIMB 9867 (strain P25X) produces isofunctional enzymes of the gentisate pathway that enables the degradation of xylenols and cresols via gentisate. Previous reports had indicated that one set of enzymes is constitutively expressed whereas the other set is strictly inducible by aromatic hydrocarbon substrates. The gene encoding gentisate 1,2-dioxygenase (GDO), the enzyme that catalyzes the cleavage of the gentisate aromatic ring, was cloned from strain P25X. The GDO gene, designated xlnE, is 1,044 bp, and is part of a 5.4 kb operon which consists of six genes, xlnEFGHID. Transcription of this operon was driven by a sigma 70-type promoter, PxlnE, located 123 bp upstream of the xlnE start codon. Primer extension analysis showed that the xlnE transcription start point is located at the -87 adenine residue. In a P25X xlnE knockout mutant, GDO activity could only be detected when cells were grown in the presence of aromatic substrates, suggesting that xlnE encodes for the constitutive copy of GDO. This was verified by constructing a P25X strain with xlnE transcriptionally fused to a promoterless catechol 2,3-dioxygenase gene. In this strain, catechol 2,3-dioxygenase activity was detected in cells that were grown in the absence of aromatic inducers. However, catechol 2,3-dioxygenase activity increased up to four fold when these cells were grown in the presence of aromatic substrates, in particular 3-hydroxybenzoate. Thus, xlnE is in fact, inducible and the constitutive activity observed under non-inducing conditions was due to its relatively high basal levels of expression. PMID- 12909359 TI - Three CCAAT-boxes and a single cell cycle genes homology region (CHR) are the major regulating sites for transcription from the human cyclin B2 promoter. AB - Cyclins are essential regulators of the cell division cycle. Cyclin B associates with the cyclin-dependent kinase 1 (cdc2) to form a complex which is required for cells to undergo mitosis. In mammalian cells three B-type cyclins have been characterised, cyclin B1, B2 and B3. The cell cycle-dependent synthesis of cyclin B1 and B2 has been investigated in detail displaying maximum expression in G2 which is mainly regulated on the transcriptional level. We have previously shown that this regulation of the mouse cyclin B2 promoter is controlled by a cell cycle-dependent element (CDE) and the cell cycle genes homology region (CHR). Also in a number of other genes CDE/CHR elements repress transcription in G0 and G1 and lead to relief of repression later during the cell cycle. Here, we compare human and mouse cyclin B2 promoters. Both promoters share only nine regions with nucleotide identities. Three of these sites are CCAAT-boxes spaced 33 bp apart which can bind the NF-Y transcriptional activator. NF-Y binding to the human cyclin B2 promoter could be shown by chromatin immunoprecipitation (ChIP) assays. Activation by NF-Y is responsible for more than 93% of the total promoter activity as measured by cotransfecting a plasmid coding for a dominant-negative form of NF-YA. Cell cycle-dependent repression is regulated solely through a CHR. Surprisingly, in contrast to the mouse promoter the CHR in the human cyclin B2 promoter does not rely on a CDE site in tandem with it. Together with the recently described mouse cdc25C promoter, human cyclin B2 is the second identified gene which solely requires a CHR for its cell cycle regulation. PMID- 12909361 TI - Type IV pilus gene homologs pilABCD are required for natural transformation in Actinobacillus actinomycetemcomitans. AB - Some clinical isolates of the gram-negative periodontal pathogen Actinobacillus actinomycetemcomitans are naturally competent for DNA uptake. In this study, we examined the sequence and the function of a type IV pilus-like pilABCD gene cluster and its downstream region in a naturally transformable A. actinomycetemcomitans strain D7S. Specific knockout mutants of pilABCD of strain D7S were constructed by replacing individual genes with an antibiotic resistance cassette. The transformation frequency of chromosome markers in the wildtype strain D7S was approximately 10(-3) per CFU. In contrast, the delta pilA, delta pilB, delta pilC, delta pilBC or delta pilD mutants were non-transformable (transformation frequency <10(-8)). Disruption of an ORF downstream of pilD had no apparent effect on the transformability of this bacterium. The pilA or pilBC deletion did not seem to affect fimbria expression or cell surface structure in either rough or smooth strains as determined by scanning and transmission electron microscopy examinations. RT-PCR analysis showed that pilA was expressed in strain D7S under a competence-inducing growth condition. The expression of pilA was barely detectable in strain D7S cultured under a non-competence-inducing condition or in the non-transformable strain JP-2. The results indicate that pilABCD are required for competence but are apparently not involved in fimbria expression of A. actinomycetemcomitans. PMID- 12909362 TI - In polymorphic genomic regions indels cluster with nucleotide polymorphism: Quantum Genomics. AB - Previously, we have described polymorphic frozen blocks (PFBs) within the Major Histocompatibility Complex (MHC) as regions of several hundred kilobases characterised by high nucleotide diversity, little or no recombination, duplicated segments, disease susceptibility, and human endogenous retroviruses. The nucleotide diversity profile within these PFBs shows peaks and troughs outside of the Class I genes, reflecting other important genes (or sequences) in the region. Here we show that indel density is also clustered with similar peaks and troughs. In fact, SNPs and indels are co-located within PFBs. PMID- 12909363 TI - Mouse SCO-spondin, a gene of the thrombospondin type 1 repeat (TSR) superfamily expressed in the brain. AB - SCO-spondin is specifically expressed in the subcommissural organ (SCO), a secretory ependymal differentiation lining the roof of the third ventricular cavity of the brain. When released into the cerebro-spinal fluid (CSF), SCO spondin aggregates and forms Reissner's fiber (RF), a structure present in the central canal of the spinal cord. SCO-spondin belongs to the superfamily of proteins exhibiting conserved motifs called TSRs for 'thrombospondin type 1 repeats' and involved in axonal pathfinding during development. The mouse SCO spondin coding sequence was searched by alignement of the coding bovine SCO spondin sequence with the mouse whole genome shotgun (WGS) supercontig (NW 000250). Compared to the bovine, mouse SCO-spondin shows 66.8% identity of amino acids. This extracellular matrix glycoprotein has a modular arrangement of several conserved domains including 25 TSRs, 10 low-density lipoprotein receptor (LDLr) type A repeats and cystein-rich regions in the -NH2 and -COOH ends. The spatio-temporal expression of SCO-spondin was analyzed using specific antisera and an homospecific SCO-spondin riboprobe. In the adult, the patterns obtained by in situ hybridization (ISH) and immunohistochemistry correlated well in the SCO, while Reissner's fiber and the ampulla caudalis were immunoreactive only. In the fetus, both the immuno and ISH reactions appeared between 14 and 15 days post coitum (dpc) in the SCO anlage. In addition, the mouse SCO-spondin gene was located at chromosome 6, between marker D6Mit352 and D6Mit119, in a conserved syntenic region. PMID- 12909365 TI - Characterization of the human gamma-glutamyl hydrolase promoter and its gene expression in human tissues and cancer cell lines. AB - Human gamma-glutamyl hydrolase (hGH) plays an important role in the metabolism of folic acid and the pharmacology of antifolates such as methotrexate. We have previously cloned and characterized hGH cDNA and its gene. We report here that the levels of hGH gene expression are high in tissues such as liver, kidney, and placenta as determined by Northern blot and RT-PCR analyses. In contrast, hGH expression is relatively low in spleen, lung, small intestine, and peripheral blood leukocytes. In addition, high levels of hGH mRNA were detected in most cancer cell lines examined. There was no significant difference in hGH mRNA levels between breast cancer tissues and their normal counterparts, although breast cancer tissues generally appeared to have heterogeneous expression of hGH mRNA. Deletion analysis and transient transfection assays were performed. A positive regulatory element located from -52 to +4 relative to the transcriptional start site was found to be required for basal promoter activity in both HepG2 and MCF-7 cells. Also, transcriptional inhibitory elements were found at -96 to-52 and +88 to+104 in MCF-7 cells, but not in HepG2 cells. These data provide novel insights into the regulation of hGH gene transcription in HepG2 and MCF-7 cells. PMID- 12909364 TI - Characterization of Drosophila palmitoyl-protein thioesterase 1. AB - Batten disease or neuronal ceroid lipofuscinoses (NCL) are a group of genetic neurodegenerative diseases that primarily afflict infants and children and are characterized by progressive loss of brain functions caused by the death of central nervous system (CNS) neurons. The most severe form of the disease is infantile NCL (INCL). INCL is caused by mutations in the palmitoyl-protein thioesterase 1 (PPT1) gene, which encodes a palmitoyl-protein thioesterase 1 enzyme that cleaves long-chain fatty acids from S-acylated proteins within the lysosome. How the loss of this activity causes the death of CNS neurons is not known. A PPT1 homolog and palmitoyl-protein thioesterase 1 enzyme activity were characterized in Drosophila melanogaster as an initial step in developing Drosophila as a model system for studying the etiology of INCL. Predicted gene CG12108 in region 8A2 of the X chromosome is 55% identical and 72% similar to human PPT1 and contains conserved catalytic residues and sites of glycosylation. Northern-blot hybridizations revealed a major 1.5 kb CG12108 transcript in unfertilized eggs, embryos, larvae, pupae, adult head and thorax, ovary, testis, and S2 tissue culture cells, as well as several minor mRNA species in some tissues. Levels of the 1.5 kb transcript were fairly uniform among tissues except in testis, where the transcript was enriched 5-fold. The same tissues also contained palmitoyl-protein thioesterase 1 enzyme activity measured using the fluorometric substrate 4-methylumbelliferyl-6-thiopalmitoyl-beta-D-glucoside. Enzyme activity was highest in testis and varied among the other tissues to a greater extent than did CG12108 message, suggesting that CG12108 is subjected to post-transcriptional regulation. Finally, flies homozygous for a deletion that removes CG12108 and three unrelated neighboring genes had less than 3% of wildtype levels of enzyme activity, consistent with CG12108 encoding functional palmitoyl-protein thioesterase 1 activity and being the fly ortholog of human PPT1. CG12108 has been appropriately renamed Ppt1. PMID- 12909366 TI - Polymerase chain reaction amplification of the Bag320 satellite family reveals the ancestral library and past gene conversion events in Bacillus rossius (Insecta Phasmatodea). AB - Polymerase chain reaction amplifications of genomic DNA in 17 individuals of bisexual and parthenogenetic populations of three subspecies of Bacillus rossius (Insecta Phasmatodea) revealed that the species still harbours the whole variability of the ancestral Bag320 satellite family, since monomers of all non hybrid Bacillus taxa plus private sequences occur in it. Bag320 monomers had not been rescued as a major satellite component in B. rossius, but possibly represent the remnant of a set of diverging sequences present in the Bacillus ancestor. Following the library hypothesis, these monomer variants have been differently amplified along the evolutionary pathways leading to present taxa in agreement with the mitochondrial phylogeny of the genus. The putative converted tracts observed are explained as the results of past gene conversion events. PMID- 12909367 TI - Functional mapping of cannabinoid receptor homologs in mammals, other vertebrates, and invertebrates. AB - Over the past decade, several putative homologs of cannabinoid receptors (CBRs) have been identified by homology screening. Homology screening utilizes sequence alignment search engines to recognize homologs. We investigated these putative CBR homologs further by 'functional mapping' of their deduced amino acid sequences. The entire pharmacophore of a CBR has not yet been elucidated, but point-mutation studies have identified over 20 amino acid residues that impart CBR specificity for ligand recognition and/or signal transduction. Twenty point mutation studies were used to construct a CBR functionality matrix. Sixteen putative CBR homologs were then mapped over the matrix. Several putative homologs did not hold up to this analysis: human GPR3, GPR6, GPR12, and Caenorhabditis elegans C02H7.2 expressed a series of crippling substitutions in the matrix, strongly suggesting they do not encode functional CBRs. Mapping the contested leech (Hirudo medicinalis) CBR sequence suggests that it encodes a functional CB1; it expresses fewer substitutions than the sea squirt (Ciona intestinalis) CB1 sequence. Mapping a putative CB2 ortholog in the puffer fish (Fugu rubripes T012234) suggests it may encode a CBR other than CB2. These findings are consistent with the lack of experimental data proving these putative CBRs have affinity for cannabinoid ligands. Matrix analysis also reveals that SR144528, a 'CB2-specific' synthetic antagonist, has affinity for non-mammalian CB1 receptors, and that L3.45 appears to be CB2-specific, its cognate in CB1 receptors is F3.45. In conclusion, functional mapping, utilizing point-mutation studies, may improve the specificity of homology screening performed by sequence alignment search engines. PMID- 12909368 TI - An inserted DNA fragment with plasmid features is uniquely associated with the presence of the GBSi1 group II intron in Streptococcus agalactiae. AB - The group II intron (GBSi1) identified downstream of the C5a-peptidase gene (scpB) in a subpopulation of Streptococcus agalactiae isolates is a suggested marker for a separate genetic lineage of serotype III isolates. In the present study two additional copies of GBSi1, one of which not previously described, were identified among serotype III isolates. All intron copies shared a common target site motif. A single copy of GBSi1 was found in a subgroup of serotype II and V isolates. In these isolates, the intron had inserted downstream of scpB, which suggests that this is the primary insertion site for GBSi1. Most bacterial group II introns described to date reside in transposable elements. The scpB locus was found to be flanked by insertion sequences similar to what has been described in an intronless serotype Ia isolate. However, this region contained an additional 2.1 kb DNA fragment present only in intron carrying isolates. This DNA fragment contained a partial transposase and putative plasmid related proteins. This may suggest that GBSi1 once was brought into the S. agalactiae genome by an integrated plasmid. PMID- 12909369 TI - Isolation of new splice isoforms, characterization and expression analysis of the human septin SEPT8 (KIAA0202). AB - SEPT8 (KIAA0202) is a member of the highly conserved septin family. Septins are membrane-associated GTPases which are involved in cytokinesis and cellular morphogenesis. Using the yeast two-hybrid system and the glutathione-S transferase pull-down assay we previously had identified the SEPT8 (KIAA0202) as interaction partner of the human septin SEPT5 (cell division cycle related-1, CDCrel-1). Since the complete cDNA sequence of the human septin SEPT8 (KIAA0202) was not known at that time, we isolated new 5' and 3' cDNA sequence of SEPT8 (KIAA0202) by screening three different cDNA libraries. In addition, we performed the characterization of SEPT8 (KIAA0202) and identified new splice variants of SEPT8 (KIAA0202). The expression pattern of SEPT8 (KIAA0202) and its interaction partner SEPT5 (CDCrel-1) is illustrated. PMID- 12909370 TI - A novel gene trap method using terminator-REMI and 3' rapid amplification of cDNA ends (RACE) in Dictyostelium. AB - We describe a novel restriction enzyme-mediated integration (REMI) method for gene trapping in Dictyostelium based on the use of a terminator-deficient vector. The vector has a blasticidin deaminase (bsr) gene as a selectable marker but lacks a terminator containing a poly(A) addition signal (AATAAA). Thus, the vector was expected to integrate into the coding region of a gene to create a fusion transcript flanked by the 3' proximal region of the trapped gene. The trapped gene can be identified by simply amplifying the fusion transcript by 3' rapid amplification of cDNA ends (3'-RACE). In the analysis of 35 integration events into known genes, the vectors were found to be integrated 20 times in close proximity to the 3' ends of the genes and in the direction of transcription. This strictly localized insertion seemed to be mediated by negative selection via the surveillance system referred to nonsense-mediated mRNA decay. In contrast, in 15 events the vector integrated in the opposite direction to transcription and at random positions throughout the coding sequence. Analysis of the trapped 3' sequences showed that the transcription of the fusion gene terminated prematurely without the apparent use of an endogenous terminator; nevertheless the transcript did exhibit a poly(A) tail. Based on these results, we designated the method terminator-REMI. Using this method, we have generated a library of tagged Dictyostelium clones from which we have thus far isolated 242 developmental mutants. PMID- 12909371 TI - Cloning and characterization of canine SHARP1 and its evaluation as a positional candidate for canine early retinal degeneration (erd). AB - Canine early retinal degeneration (erd) is an early onset form of canine progressive retinal atrophy phenotypically similar to human retinitis pigmentosa. In a previous study, the locus responsible for erd was mapped to canine chromosome 27 in the region corresponding to HSA12p, a region where no human retinal degeneration loci have been mapped. Canine SHARP1 gene has been localized on CFA27 in the erd interval by RH mapping, and considered as a positional candidate gene for erd. SHARP1 was cloned and sequenced from normal and erd affected dogs, and no disease-causing mutations were identified. Genotyping of 117 dogs from informative pedigrees did not reveal any recombinants between SHARP1 and erd. To date SHARP1 gene is the closest gene-specific marker to erd; genotyping additional informative pedigrees, and sequencing SHARP1 upstream regions from normal and affected dogs will be necessary to establish if SHARP1 is involved in this canine retinal disease. PMID- 12909373 TI - The Resuscitation greats. Claude Bernard: on the origin of carbon monoxide poisoning. PMID- 12909374 TI - The epidemiology of cardiac arrest in children and young adults. AB - AIM: To describe the epidemiology of children and young adults suffering from out of-hospital cardiac arrest. PATIENTS: All patients suffering from out-of-hospital cardiac arrest in whom, resuscitation efforts were attempted in the community of Goteborg between 1980 and 2000. METHODS: Between 31 October 1980 and 31 October 2000, all consecutive cases of cardiac arrest in which the emergency medical service (EMS) system responded and attempted resuscitation were reported and followed-up to discharge from hospital. RESULTS: Among 5505 cardiac arrests information on age was available in 5290 cases (96%). Of these 5290 cases 98 (2%) were children (age 0-17 years), 197 (4%) were young adults (age 18-35 years) and the remaining 4995 (94%) were adults (age >35 years). Children and young adults differed from adults by suffering from a witnessed arrest less frequently, being found by the ambulance crew in ventricular fibrillation/tachycardia less frequently and being judged as having an underlying cardiac aetiology less frequently. Of the children only 5% were discharged from hospital alive compared with 8% for young adults and 9% for adults. Among survivors the cerebral performance categories (CPC) score at discharge tended to differ with 38% of young adults registering a CPC score of 1 (no neurological deficit) compared with 52% among adults. CONCLUSION: Children and young adults suffering from out-of hospital cardiac arrest differed from adults in terms of aetiology and observed initial arrhythmia. Children had a particularly bad outcome whereas young adults had a similar outcome as adults. PMID- 12909375 TI - An evaluation of the PA(Xpress) pharyngeal airway - a new single use airway device. AB - The PA(xpress) phayngeal airway (PA(x)) is a new single use airway device that might be used for airway maintenance during anaesthesia or cardiopulmonary resuscitation. We evaluated the performance of the PA(x) in 103 anaesthetised non paralysed patients undergoing non-emergency anaesthesia. We recorded success of insertion, quality of airway achieved and complications of its use. We were successful in establishing a clear airway on the first attempt on 68 (67%) occasions. We were unable to establish a patent airway in nine (9%) patients. Partial or intermittent airway obstruction occurred during maintenance of anaesthesia in ten (11%) cases but none required removal of the device. Jaw thrust was used to assist insertion in all cases. A mean of 1.37 further manipulations per patient were required to establish an airway and 0.60 per patient were required during maintenance of anaesthesia. In the 94 patients in whom an airway was established, assisted ventilation was excellent in 77 (82%). Leak pressure was 20 cmH2O or above in 58% cases. Intracuff pressure was measured in 55 patients: mean pressure was 68 cmH2O and was above 100 cmH2O in ten (18%) cases. Complications occurred in 38 (37%) patients during insertion, in a further 12 (13%) during maintenance and in eight (9%) during emergence. The device was difficult to insert and associated with a high incidence of trauma: blood was visible on the device after removal in 56 (55%) cases. We conclude that the PA(x) is associated with too high a failure rate and too high an incidence of minor complications for routine airway maintenance. PMID- 12909376 TI - Diagnosis and management of out-of-hospital cardiac arrest secondary to coronary artery spasm. AB - OBJECTIVE: The clinical features of coronary artery spasm as a cause of cardiac arrest were determined in a prospective study on out-of-hospital cardiac arrest (OHCA). METHODS: Coronary angiography was performed at admission in 300 consecutive patients with no obvious non-cardiac cause of OHCA. In survivors with no or minimal coronary artery stenosis, a second angiography with provocation test and electrophysiological testing were performed at 1 month. RESULTS: Spasm was demonstrated in ten patients. Diagnosis was based upon (1) spontaneous spasm on the admission angiogram (3 patients), (2) transient significative ST-segment elevation at follow-up in patients with no or non-significant coronary artery lesions (4 patients) and (3) spasm during the 1 month provocation test (3 patients). Six patients survived at 1 month; spasm occurred during a new provocation test in five despite treatment with high dosage calcium channel blockers leading to coronary stenting in two, an internal cardiovertor defibrillator in one, and increased drug therapy with prolonged hospitalization in the remainder. At a mean follow-up of 55+/-27 months, no recurrent cardiac arrest occurred. CONCLUSION: Systematic coronary angiograms and provocation tests in survivors of OHCA allow prompt diagnosis of coronary artery spasm. Residual spasm despite treatment with calcium channel blockers is frequent. Therapy should therefore be guided by repetitive provocation tests, and seems to avoid recurrence of cardiac arrest. PMID- 12909377 TI - A review of emergency equipment carried and procedures performed by UK front line paramedics. AB - OBJECTIVES: To assess which items of resuscitation equipment are carried on United Kingdom (UK) front line ambulances and what procedures paramedics are able to perform. To compare these findings with those of a previous survey in 1997. METHOD: Postal survey to the chief executives of all the UK ambulance services and direct comparison with the data from 1997. RESULTS: Nasopharyngeal airway usage (NPA) has increased (21-55%) and the laryngeal mask airway (LMA) (from 10 to 26%). No services employ the Combitube. 94% of services use a Hudson type trauma mask (increase of 17%). One service no longer allows its paramedics to deliver 12-15 l of oxygen. 68% of trust paramedics can perform needle thoracocentesis (increase of 35%). No paramedics perform chest drain insertion. All services have 14G intravenous cannulae available and 45% carry the intraosseous needle (increase of 30%). No services employ the MAST suit. There is an increase by 29% in the use of crystalloids and a decrease in the use of colloids of 22%. 23% of fluid resuscitation protocols are based upon the principles of hypotensive resuscitation. Spinal boards and extrication devices are used by 97% of services. The use of inflatable splints has decreased (38 23%). There has been a minor increase in the use of traction splints to 74% of services. The use of Entonox is universal. Nalbuphine (Nubain) is the most widely used opioid. The use of morphine/diamorphine has doubled to 10% with a further 26% to introduce their use. 29% of services have equipped some vehicles with automatic external defibrillators. CONCLUSIONS: The equipment available to UK paramedics and procedures that they may perform continues to expand. There are still variations in the basic management of airway, breathing and circulation care and only some services are keeping up to date with current medical thinking, for example the increasing use of crystalloids and hypotensive resuscitation. It remains to be seen whether the widespread use of Nalbuphine as a first line analgesic may decrease as the use of natural strong opiates becomes more widespread. PMID- 12909379 TI - Cardiac arrest in private locations: different strategies are needed to improve outcome. AB - BACKGROUND: A tremendous amount of public resources are focused on improving cardiac arrest (OHCA) survival in public places, yet most OHCAs occur in private residences. METHODS AND RESULTS: A prospective, observational study of patients transported to seven urban and suburban hospitals and the individuals who called 911 at the time of a cardiac arrest (bystander) was performed. Bystanders (N=543) were interviewed via telephone beginning 2 weeks after the incident to obtain data regarding patient and bystander demographics, including cardiopulmonary resuscitation (CPR) training. Of all arrests 80.2% were in homes. Patients who arrested in public places were significantly younger (63.2 vs. 67.2, P<0.02), more often had an initial rhythm of VF (63.0 vs. 37.7%, P<0.001), were seen or heard to have collapsed by a bystander (74.8 vs. 48.1%, P<0.001), received bystander CPR (60.2 vs. 28.6%, P<0.001), and survived to DC (17.5 vs. 5.5%, P<0.001). Patients who arrested at home were older and had an older bystander (55.4 vs. 41.3, P<0.001). The bystander was less likely to be CPR trained (65.0 vs. 47.4%, P<0.001), less likely to be trained within the last 5 years (49.2 vs. 17.9, P<0.001), and less likely to perform CPR if trained (64.2 vs. 30.0%, P<0.001). Collapse to shock intervals for public versus home VF patients were not different. CONCLUSIONS: Many important characteristics of cardiac arrest patients and the bystander differ in public versus private locations. Fundamentally different strategies are needed to improve survival from these events. PMID- 12909378 TI - First aid knowledge of alpine mountaineers. AB - OBJECTIVE: The study evaluates the knowledge of first aid in mountaineerers who climb routes of moderate difficulty in the western Alps. Additionally the mountaineer's ability to assess their own knowledge was investigated. An analysis of the mountain accidents in the same area showed the real need for knowledge. DESIGN: An investigation of a cohort of mountaineers who reached Margherita Hut (4559 m, Monte Rosa; n=283; 17 questions with five answers each (11 themes)). RESULTS: Knowledge in general is poor. Best results were obtained in relation to cardiac emergencies, altitude sickness, and hypovolaemic shock, and worst resulted with hypothermia, traumatic injuries, treatment of pain and management of emergencies. Although traumatic injuries represent about 50% of mountain accidents in the region, there was a general lack of basic knowledge on this subject. Self-assessment of the individuals level of knowledge and their need for further education was inadequate. Differences between sex, age, nationality, mountaineering professional experience (medical education) are discussed. CONCLUSION: First aid education of mountaineers must be improved. Adequate education should take into account the specific demands of alpine emergencies. PMID- 12909380 TI - A randomized controlled trial of chest compression only CPR for older adults-a pilot study. AB - Older people are trained infrequently in cardiopulmonary resuscitation (CPR), yet are more likely to witness a cardiac arrest. Older people who are CPR trained perform CPR when witnessing a cardiac arrest. OBJECTIVE: To assess whether elderly adults (>55 years) who receive chest-compression only cardiopulmonary resuscitation (CC-CPR) training display equivalent skill retention rates compared with those who receive traditional CPR instruction. We also identified factors associated with 3 months skill retention at 3 months in both groups. METHODS: Older adults in a suburban hospital Older Adult Services program were invited to participate in an experimental CPR course. The 2 h course was modelled after the AHA Friends and Family course, and used one of two standardized video scenarios. Seventy four subjects were randomized to CC-CPR (n=36) or traditional CPR (n=38) training. Participation consisted of initial training, followed by a 3 months return videotaped assessment. Three months skill competence was assessed either by consensus between two video evaluators, or the on-site evaluator. Chi square and Kappa tests were used for analysis, and unadjusted odds ratios and 95% confidence intervals are reported. RESULTS: Skill retention assessments were completed on 29 (81%) CC-CPR and 26 (68%) CPR trainees. Subjects were elderly (71.5+/-6.69 years), and had a high rate of previous CPR training (58.0%). Groups were similar in demographic characteristics. After training, participants exhibited high rates of perceived competence (86.4%), although the overall 3 months skill retention was low (43.6%). CC-CPR training resulted in equivalent skill retention rates as compared with traditional CPR training (51.7 vs. 44.4%; P=0.586). No participant factors were associated with skill retention, including age, previous CPR training, education level, medical history, or perceived physical ability to perform. CONCLUSION: We identified low rates of CPR skill retention in this elderly population. CC-CPR instruction was associated with equivalent skill retention rates compared with traditional CPR instruction. No demographic factors were associated with successful skill retention. PMID- 12909381 TI - Knowledge of basic life support: a pilot study of the Turkish population by Baskent University in Ankara. AB - A total of 494 participants who were scheduled to take Baskent University's basic life support (BLS) training programme in 2001-02 were asked to complete a 25-item questionnaire prior to the course. The questionnaire investigated the demographic characteristics of the subjects, their knowledge of the theoretical and practical aspects of BLS, and personal experience and attitudes related to BLS. The 'non medical group' included 179 laypersons, and the 'medical group' was composed of medical students (n=220), residents (n=69) and clinical nurses (n=26). One hundred and twenty-six (25.5%) of the participants had an encounter with some form of medical emergency event in their past and 207 (41.9%) had taken a BLS course previously. The most commonly cited anxiety about performing BLS was the fear of further harming the victim (56.9%). Nine of the participants said they would not perform mouth-to-mouth ventilation (1.8%). Compared to the other participants, individuals with previous emergency experience, and those who had previous BLS training answered significantly more of the theoretical questions correctly. However, neither of these groups performed significantly better than the other participants in the practical questions (P>0.05 for both comparisons). Based on our findings, we recommend that BLS training for medical undergraduates, other medical personnel and laypersons be improved and standardized throughout Turkey. PMID- 12909382 TI - Quality of CPR with three different ventilation:compression ratios. AB - Current adult basic cardiopulmonary resuscitation (CPR) guidelines recommend a 2:15 ventilation:compression ratio, while the optimal ratio is unknown. This study was designed to compare arterial and mixed venous blood gas changes and cerebral circulation and oxygen delivery with ventilation:compression ratios of 2:15, 2:50 and 5:50 in a model of basic CPR. Ventricular fibrillation (VF) was induced in 12 anaesthetised pigs, and satisfactory recordings were obtained from 9 of them. A non-intervention interval of 3 min was followed by CPR with pauses in compressions for ventilation with 17% oxygen and 4% carbon dioxide in a randomised, cross-over design with each method being used for 5 min. Pulmonary gas exchange was clearly superior with a ventilation:compression ratio of 2:15. While the arterial oxygen saturation stayed above 80% throughout CPR for 2:15, it dropped below 40% during part of the ventilation:compression cycle for both the other two ratios. On the other hand, the ratio 2:50 produced 30% more chest compressions per minute than either of the two other methods. This resulted in a mean carotid flow that was significantly higher with the ratio of 2:50 than with 5:50 while 2:15 was not significantly different from either. The mean cerebrocortical microcirculation was approximately 37% of pre-VF levels during compression cycles alone with no significant differences between the methods. The oxygen delivery to the brain was higher for the ratio of 2:15 than for either 5:50 or 2:50. In parallel the central venous oxygenation, which gives some indication of tissue oxygenation, was higher for the ratio of 2:15 than for both 5:50 and 2:50. As the compressions were done with a mechanical device with only 2 3 s pauses per ventilation, the data cannot be extrapolated to laypersons who have great variations in quality of CPR. However, it might seem reasonable to suggest that basic CPR by professionals should continue with ratio of 2:15 at present if it can be shown that similar brief pauses for ventilation can be achieved in clinical practice. PMID- 12909383 TI - Effect of amiodarone on haemodynamics during cardiopulmonary resuscitation in a canine model of resistant ventricular fibrillation. AB - OBJECTIVE: Amiodarone has been shown to be superior to both placebo and lidocaine in improving survival to hospital admission for victims of out-of-hospital refractory ventricular fibrillation. Concern had been expressed about the known vasodilatatory effects of amiodarone if given without precedent vasoconstrictive medications. The haemodynamic effects of intravenous amiodarone administered during ongoing CPR have not been systemically investigated. Our intention was to verify if amiodarone alone produced significantly lower resuscitation haemodynamics than did either adrenaline (epinephrine) alone or the combination of amiodarone and adrenaline. DESIGN: Prospective, randomized, comparative study. SETTING: Research laboratory of a medical school. SUBJECTS: Thirty mongrel dogs. INTERVENTIONS: After 8 min of untreated VF, defibrillation was attempted once at 3 J/kg and external chest compressions and ventilation started. Those animals resistant to the defibrillation attempt were randomized, ten to an adrenaline (0.02 mg/kg) group, ten to an amiodarone (5 mg/kg) group, and ten to a group receiving a combination of both drugs. MEASUREMENTS AND MAIN RESULTS: Aortic systolic and diastolic, and coronary perfusion pressures were all significantly lower in the group receiving amiodarone alone than in the other two groups. Amiodarone combined with adrenaline produced pressures during CPR similar to adrenaline alone. CONCLUSION: Amiodarone can be safely administered simultaneously in combination with adrenaline and such a combination results in similar haemodynamic support as adrenaline alone. Amiodarone administered alone produces significantly lower coronary perfusion pressure than when combined with adrenaline. PMID- 12909384 TI - Intestinal ischaemia during cardiac arrest and resuscitation: comparative analysis of extracellular metabolites by microdialysis. AB - Intestinal ischaemia is a major complication of shock syndromes causing translocation of bacteria and endotoxins and multiple organ failure in intensive care patients. The present study was designed to use microdialysis as a tool to monitor intestinal ischaemia after cardiac arrest and resuscitation in pigs. For this purpose, microdialysis probes were implanted in pig jejunal wall, peritoneum, skeletal muscle and brain, and interstitial fluid was obtained during circulatory arrest (induced by ventricular fibrillation) and after return of spontaneous circulation (ROSC). Cardiac arrest for 4 min caused a prolonged (60 min) reduction of blood flow in jejunal wall, muscle and brain as determined by the ethanol technique. This was accompanied by cellular damage in heart muscle and brain as indicated by increased levels of troponin-I and protein S-100, respectively. Plasma levels of glucose, lactate and choline were increased at 15 60 min following cardiac arrest. In contrast, cardiac arrest induced a rapid but variable decrease of interstitial glucose levels in all monitored organs; this decrease was followed by an increase over baseline during reperfusion. In the intestine, lactate, glutamate and choline levels were increased during ischaemia and reperfusion for 60-120 min; intestinal and peritoneal samples yielded parallel changes of lactate levels. Brain and muscle samples showed similar changes as in intestinum and peritoneum except for glutamate, which was increased in brain but not in muscle. We conclude that intestinal ischaemia occurs as a consequence of cardiac arrest and resuscitation and can be monitored by in vivo microdialysis. Comparative analysis by multi-site microdialysis reveals that the intestine is equally or even more sensitive to ischaemia than brain or muscle. PMID- 12909386 TI - Non-specific hyperamylasemia in shosin beri-beri. AB - Several reports demonstrate non-specific hyperamylasemia in cardiac surgery or diabetic ketoacidosis. We report here for the first time non-specific hyperamylasemia in a cardiovascular beri-beri case who showed shock with severe metabolic acidosis. Her echocardiography revealed hyperkinetic wall motion of the small left ventricle. Despite intravascular volume expansion in parallel with dopamine administration, her blood pressure did not recover. Abdominal computed tomography (CT) did not reveal pancreatic swelling or any other signs of acute pancreatitis. Her history suggested a possibility of cardiovascular beri-beri due to chronic alcoholism. Thiamine administration dramatically reversed her haemodynamic derangements, metabolic acidosis and even relieved her abdominal pain. Isozyme examinations for hyperamylasemia showed that most of the serum amylase consisted of salivary type. This case report expands our information on non-specific hyperamylasemia encountered in the emergency setting. PMID- 12909385 TI - Gas gangrene pyaemia with myocardial abscess formation--fatal outcome from a rare infection nowadays. AB - We report a case of sudden death after gas gangrene. A 67-year-old male patient with diabetes mellitus and chronic renal failure (on haemodialysis three times a week) presented in the surgical emergency department with a severe swelling and crepitation in the right groin. No signs of trauma were present-except for a well healed, 1-year-old scar after femoro-popliteal bypass surgery. Two days earlier, he had presented to the internal medicine department with epigastric pain and had left against medical advice. On readmission the patient was initially conscious and in a stable cardiopulmonary condition but developed sudden cardiocirculatory failure and underwent resuscitation. Despite all resuscitation measures, including the administration of high doses of catecholamines and the treatment of hyperkalemia, the patient died. Autopsy revealed septicaemia with rod-shaped gram positive bacteria, typical of Clostridium perfringens, evidenced by multiple areas of myonecrosis. Abscess formation was found in the myocardium. Clostridial gas gangrene is a rare clinical condition. Unless immediate diagnosis and adequate therapy measures are taken, the outcome and chances for survival are poor as demonstrated by this case. PMID- 12909387 TI - Implication of epinephrine-induced hypokalemia during cardiac arrest. PMID- 12909389 TI - [ERPs (N200, P300 and CNV) in alcoholics: relapse risk assessment]. AB - The cognitive event-related potentials were studied in a group of 55 alcoholic patients, paired in age and sex with a group of 18 control subjects, using a protocol oddball (visual and auditory) and a protocol VCN Go/Nogo. The N200 obtained using an auditory oddball paradigm had lower amplitude in alcoholics than in controls. A significant amplitude decrease of visual P300 was also observed in alcoholic male subjects. There was however no difference in auditory P300 between alcoholics and control subjects. Using a Go/Nogo paradigm, a significant difference on the final part of the VCN appears between alcoholic and pilot subjects. In addition, the longitudinal follow-up of the same alcoholic patients showed an electrophysiological profile that allowed dividing them into two different groups. On the initial recording (17 days after weaning), the auditory oddball P300 amplitude was significantly higher at Cz and Pz among patients who relapsed during the 3 months follow-up. The same effect appeared on the CNV protocol, where the amplitude of P300 was higher in patients who subsequently relapsed than for those who remained abstinent. Cognitive ERPs may be clinically useful to improve the prediction of risk of relapse among alcoholic patients. PMID- 12909390 TI - [EEG and ischemic stroke in full-term newborns]. AB - The aims of this study were to describe EEG anomalies in unilateral neonatal ischemic stroke without hypoxic-ischemic encephalopathy, and to determine possible links between these abnormalities and long-term outcome. In 6 full-term newborns without severe fetal distress ischemic stroke was confirmed by computed tomography and/or magnetic resonance imaging. Twenty EEGs were recorded during the neonatal period, 5 in acute stage and 15 later. The duration of the follow-up ranged from 3 to 9 years. All newborns developed unilateral clonic seizures, right-sided (5 cases) or left-sided (1 case); seizures began between 14 and 48 h of life. At follow-up, 3 children were normal at 2 and 6 years of age, while the 3 others had sequelae: epilepsy at 9 years of age in one, and unilateral mild cerebral palsy in the 2 others (3 and 4 years of age), with behavioral problems in one of them. Critical EEG discharges, rhythmic sharp waves and/or slow waves were recorded on the injured side. Abnormalities of interictal activity were excess of alpha or theta rhythms, transitory EEG discontinuity or low voltage. The 2 children with cerebral palsy had numerous unilateral post-ictal positive rolandic slow sharp waves (PRSSWs), which were similar to the positive rolandic sharp waves of premature infants; the child with behavioral problems had numerous positive left-sided temporal fast sharp waves. PRSSWs could be associated with contralateral motor sequelae, while positive left temporal fast sharp waves were associated with long term behavioral problems. These findings may be used for future prospective studies aimed at specifying the relation between EEG abnormalities and long-term outcome. PMID- 12909391 TI - Time course of motor excitability before and after a task-related movement. AB - AIMS OF THE STUDY: The time course of motor excitability during a task-related unilateral right thumb movement was studied using sub-threshold transcranial magnetic stimulation (TMS) to the contralateral left motor cortex. The level of stimulation evoked a motor evoked potential (MEP) in the thumb when the subject was at rest in approximately 10% of the trials. METHODS: Subjects made a brief right thumb movement to the predictable omission of regularly presented tone bursts allowing experimental definition of TMS relative to the cue to move. Motor cortical excitability was characterized by amplitude and/or probability of eliciting MEPs. RESULTS: There were four periods of altered motor excitability during task performance compared to a control resting state: a first period of weak facilitation before movement between -500 to -200 ms, a second period without increased excitability approximately 150 ms before movement onset when MEPs amplitude was below that seen in rest, a third period of strong facilitation between -100 ms before movement and +200 ms after facilitation and a fourth period of weak facilitation between +200 to +500 ms. CONCLUSION: These results show that during performance of a task requiring a motor response, motor cortical excitability is increased above resting for hundreds of millisecond before and after the response, except for a transient period between 75 and 150 ms prior to movement onset. The temporal pattern of these excitability changes is compatible with multiple excitatory and inhibitory inputs interacting on motor cortex. PMID- 12909392 TI - [Effects of a subanaesthetic dose of ketamine on emotional and behavioral state in healthy subjects]. AB - Schizophrenic patients suffer from positive (delusions, hallucinations) and negative signs (social withdrawal) as well as emotional disturbance that included quantitative (blunted affect) and qualitative impairments (discordance of emotional level). Ketamine, a phencyclidine derivative, is a non competitive N methyl-D-aspartate (NMDA) glutamate receptor antagonist. In healthy subjects its administration induces some positive symptoms (perceptual distortions.), negative symptoms (emotional deficit, apathy, social withdrawal) and cognitive changes (memory impairments and perseverations) that resemble some aspects of the symptoms of schizophrenia. A double blind cross over, placebo controlled was performed in 12 normal subjects with 2 sessions separated by one week of wash-out to determine ketamine-induced effects on behavioral and emotional responses. During each session, subjects received either ketamine or placebo (saline) infusion. A subanesthetic dose of ketamine (0,5 mg/kg) was administered by constant perfusion over 60 min. Behavioral and cognitive responses were assessed using positive and negative symptoms scales (BPRS, items from SAPS and SANS), vigilance and mood visual analog scale, subjective feelings using the Addiction Research Center Inventory (ARCI) and the Profile of Mood States (POMS). Using Philippot's method, emotions were elicited by films segments which induce a diversity of predictable emotions (fear, anger, sadness, joy, disgust and neutral state) and emotional responses were assessed by the Differential Emotions Scale (DES Izard). Low dose of ketamine induced significant effects on 7-items BPRS score (positive and negative items) and significant effects on positive and negative symptoms from SANS and SAPS. This was associated with emotional blunting of visually-induced responses that resemble aspects of schizophrenic emotional impairments. Ketamine impaired ARCI subscales (benzedrine subscale, pentobarbital chlorpromazine subscale and LSD subscale). The recent findings of ketamine's pharmacology and imaging studies allow to draw several hypothesis related to neurotransmitter systems (glutamate, dopamine, serotonin interactions) and cerebral areas (particularly prefrontal cortex, anterior cingulate cortex, hippocampus) underlying some of these ketamine-induced effects. PMID- 12909395 TI - On-line information sources of toxicology in Finland. AB - Toxicology is relatively young as an independent discipline in Finland as in most other countries. Toxicology gradually evolved at universities as a part of pharmacology during 1960-1970s. The research institutes, with the Finnish Institute of Occupational Health being a pioneer in this respect, have contributed significantly in special research areas. The current paper highlights Finnish toxicology, and concentrates on the current status of sources of digital information and their use. PMID- 12909394 TI - On-line sources of toxicological information in Canada. AB - This paper will provide an overview of the on-line resources available in toxicology in Canada. It will describe a brief history of The Society of Toxicology of Canada, with reference to other societies and also provide information on education, research and other resources related to toxicology. Toxicology in Canada emerged as a distinct and vibrant discipline following the thalidomide tragedy of the 1960s. In the pharmaceutical industry and government, toxicology was readily established as an essential component of drug development and safety, and as the need for toxicologists expanded, training programs were established, usually in collaboration with departments of pharmacology. In the last two to three decades other disciplines, environmental biology, analytical chemistry and epidemiology joined the ranks of toxicology. The on-line sources of toxicology information are rapidly expanding. This article describes those sources considered by the authors to be important from a national and international perspective. The majority of these sources are professional organizations and government agencies. PMID- 12909397 TI - Information resources in toxicology--Italy. AB - The purpose of the present paper is to provide an overview of current resources in the field of toxicology in Italy. The discussion will begin with a brief history of toxicology in this country, which includes the study of the toxicity of plants and other natural substances, and the birth of industrial and forensic toxicology. We will also provide information on research, education, and hazard control in the field of toxicology. Within this context we will examine the public bodies responsible for surveillance and regulatory activities, state-owned and private structures involved in toxicological research, and the educational programs and research activities of universities. Particular emphasis will be placed on the activities of the National Health Service, which plays an important role in areas such as clinical toxicology, food safety, and animal health, as well as those of national and regional agencies dedicated to the protection of the environment. The presentation will be organized as follows: (1) A Brief History of Toxicology in Italy; (2) Professional Societies; (3) National Health Service; (4) National Bodies; (5) Resources for the Environment; (6) Biomedical Websites; (7) Recent Publications; (8) Research Structures; (9) Graduate and Postgraduate Programs; (10) Legislation. PMID- 12909396 TI - Germany: toxicology information on the World Wide Web. AB - Following a brief introduction to the history of toxicology in Germany, this article deals with institutions concerned with toxicological information and research in Germany. The web addresses, responsibilities and main topics of federal authorities, federal state authorities, institutions for statutory accident insurance and prevention, non-university and university research institutes, funding institutions, scientific societies, industry representatives and poison information centers are described and links are given to databases provided on the web sites of these institutions. PMID- 12909398 TI - History and current state of toxicology in Russia. AB - This paper provides a brief history of toxicology and an overview of current research activities and education in toxicology in Russia. It includes a list of national bodies and research institutes, as well as laws, regulatory documents, and databases in toxicology. PMID- 12909399 TI - Online information resources of toxicology in Sweden. AB - This article presents toxicological information resources in Sweden available to the public on the Internet. The main focus is on websites of organizations and universities with toxicological information in English. For example, the National Chemicals Inspectorate (KemI) has several databases with information on chemical substances on their website as well as regulatory information about chemicals. The Swedish Environmental Protection Agency has a database containing environmental information and on the website of the National Institute of Working Life a database on occupational health is available. An important part of toxicological research is carried out at the Institute of Environmental Medicine at the Karolinska Institute. Several universities and colleges are responsible for research, education and training in toxicology and ecotoxicology. PMID- 12909400 TI - Toxicology digital sources produced and available in the United Kingdom (UK). AB - This paper looks at toxicology digital sources produced and available in the United Kingdom (UK). It includes products aggregating a number of sources, which may include information from other countries, and for the convenience of the reader and information seeker these have also been described. PMID- 12909401 TI - Global information network on chemicals (GINC) and its Asian component. AB - The Global Information Network on Chemicals (GINC) is an effort to build a global information network that links international, national, and other organizations working for the safe management of chemicals in order to exchange information and improve communications. The project was originally proposed in 1993 by one of the authors then at the National Institute of Health Sciences (NIHS) of Japan to the International Program on Chemical Safety (IPCS), which is a joint project of World Health Organization (WHO), International Labor Organization (ILO), and United Nations Environment Program (UNEP). The base support system was first implemented at NIHS using the Internet/World Wide Web (WWW) technology in 1995. The project was then endorsed by the Intergovernmental Forum on Chemical Safety (IFCS) and was adopted by the Inter-Organization Program for the Sound Management of Chemicals (IOMC). However, the base system (http://www.nihs.go.jp/GINC/index.html) has been developed and maintained solely by the NIHS group under the support of the Ministry of Health and Welfare (MHW), Japan. Asia, particularly East Asia and the Pacific region, was chosen as the feasibility study region for this project. During the period from December 1994 to July 2002, NIHS hosted eight meetings on this project held in Tokyo. PMID- 12909402 TI - ILO activities in the area of chemical safety. AB - The ILO has been active in the area of safety in the use of chemicals at work since the year of its creation in 1919, including the development of international treaties and other technical instruments, the provision of technical assistance to its member States, and the development of chemical safety information systems. The two key ILO standards in this area are the Conventions on safety in the use of chemicals at work (No. 170, 1990), and the Prevention of Major Industrial Accidents (No. 174, 1993). The ILO Programme on occupational safety, health and environment (Safe Work) is currently responsible for ILO chemical safety activities. In the past two decades, most of ILO work in this area has been carried out within the context of inter-agency collaboration frameworks linking the ILO, WHO, UNEP, FAO, UNIDO, UNITAR, and the OECD, including the International Programme on Chemical Safety (IPCS), the Inter Organisation Programme for the Sound Management of Chemicals (IOMC), and the Intergovernmental Forum on Chemical Safety (IFCS). Apart from the regular development, updating and dissemination of chemical safety information data bases such as the IPCS International Chemical Cards, the elaboration of a Globally harmonized system for the classification and labelling of Chemicals (GHS) has been the most outstanding achievement of this international collaboration on chemical safety. PMID- 12909404 TI - OECD environment, health and safety programme: information on the world wide web. AB - This article presents the computerised resources of the Environment, Health and Safety (EHS) Programme of the Organisation for Economic Co-operation and Development. The main focus is the dissemination of tools for chemical safety assessment, which are used by OECD member countries, with the aim of promoting harmonization. The contents include the activities of the programme with special emphasis on dissemination of information via the web site (http://www.oecd.org/ehs) in the form of up-to-date information on activities, publications, directories, databases, contact points and links to other sites of importance. PMID- 12909403 TI - The International Union of Toxicology (IUTOX): history and its role in information on toxicology. AB - The International Union of Toxicology (IUTOX) was founded in 1980 in Brussels. The initiative was started by the Society of Toxicology (SOT), USA, and the European Society of Toxicology in 1975 (EST), and the foundation prepared by an Inter Society Liason Committee. Eight National Societies of Toxicology were founding members of IUTOX besides SOT and EST. It now comprises 43 national/regional Societies from all over the world, representing over 20,000 toxicologists. Information has always been a key element in toxicology. Information exchange in IUTOX has evolved from letters and phone calls to fax, e mail and the use of the Internet. Initially, newsletters were created which were mailed and later displayed on the Web. The website has been developed as a major information tool with many useful links, thereby providing information for the scientific community, the media and the lay public. PMID- 12909405 TI - UNEP Chemicals' work: breaking the barriers to information access. AB - The global production and trade in chemicals was measured at 1.55 trillion US dollars in 1996, four times the amount of 1966. Chemicals represent 14% of imports and exports of OECD countries and the present decade has been characterised by the expansion of the production and use of chemicals in countries with economies in transition and developing countries. While some of these chemicals are benign at the levels we are usually exposed to them, others present risks to human health or the environment often across national boundaries. Failure to safely manage these risks could lead to serious and long term consequences. The sound management of chemicals requires adequate information about their nature and use as well as their effects and control options. Much of this information can be culled from a variety of sources but that does not guarantee equal access to it by all those who are in need. To achieve equitable information distribution, the chemical unit of the United Nations Environment Programme (UNEP) operates many activities aimed at removing the geographical and technical barriers that prevent the free flow of chemical management information. These activities include the use of assorted media and activities to enhance countries' internal capacity to search and use the information available on the Internet. PMID- 12909406 TI - Analysis of repeated tests for interferon-gamma (IFN-gamma) response and faecal excretion for diagnosis of subclinical paratuberculosis in Danish cattle. AB - A total of 315 cattle were tested for infection with Mycobacterium avium subsp. paratuberculosis (M. paratuberculosis) at three consecutive samplings, using the interferon-gamma (IFN-gamma) test on whole blood and bacteriological culture of faecal samples. Of 205 cattle from 10 infected herds 99 (48%) were positive in the IFN-gamma test on at least one sampling using "IDEXX-criteria" for interpretation, and of 110 cattle from five non-infected herds three (3%) were positive. Forty-four animals from infected and one from non-infected herds tested positive at all three samplings. Although support for the specificity of the IFN gamma test was provided by these results, they also indicate problems with false positives. Approximately half of the positive animals did not give the same result at all three samplings, indicating that repeated testing increases the chance of detecting reactors. Changing, or fluctuating, IFN-gamma test results occurred most frequently in animals younger than 1 year, indicating that the IFN gamma test should be applied only to animals 1 year and older. M. paratuberculosis was isolated from 16 (4%) of 371 cattle, all from infected herds. Fifteen culture-positive cattle tested positive at least once in the IFN gamma test. It was not possible to predict from the IFN-gamma test result the number of animals that would eventually develop disease. However, the test may be useful to detect animals that have been exposed to M. paratuberculosis earlier in their lives, and the testing of young cattle could be included in a control program to check for the effectiveness of preventing transmission of infection to calves and to identify animals at risk of developing disease later in their lives. PMID- 12909407 TI - The immunostimulatory activity and stability of grass carp (Ctenopharyngodon idellus) roe lectin. AB - A hexameric rhamnose-specific lectin with a molecular mass of 205kDa and exhibiting some N-terminal sequence similarity to other fish lectins was isolated from roe of the grass carp (Ctenopharyngodon idellus) by affinity chromatography on rhamnose-Sepharose and ion exchange chromatography by fast protein liquid chromatography on a Mono S column. The lectin exhibited mitogenic activity toward murine splenocytes with a potency lower than that of the plant lectin ConA. It exerted a stimulatory effect at a concentration of 10 micro g/ml on the phagocytic activity of seabream (Sparus sarba) macrophages. It was unstable toward heat (temperature > or =40 degrees C), acid (0.1M HCl), alkali (0.1M NaOH), trypsin and succinylation. PMID- 12909408 TI - Immune modulation following immunization with polyvalent vaccines in dogs. AB - A decline in T-cell-mediated immunity and transient state of immunosuppression after immunization has been reported in dogs. Nevertheless, dogs are still routinely vaccinated with polyvalent live vaccines and severe disease does not generally occur. In order to investigate these effects on the canine immune system and to elucidate possible mechanisms we determined the following immune parameters in the blood of 33 clinically sound German shepherd dogs before and after standard vaccination with a polyvalent vaccine against distemper, parvovirus, viral hepatitis, leptospirosis, kennel cough and rabies: white and differential blood cell count, the serum concentrations and/or activities of IL 1, IL-2, IFN-gamma, TNF-alpha, neopterin and IgG, natural killer (NK) cell activity, bactericidal activity and complement hemolytic activity, lymphocyte proliferation test (LPT) and nitroblue tetrazolium test (NBT). Our major findings were that significant postvaccinal decreases in T-cell mitogenic response to PHA and in neutrophil function and neopterin serum concentration were accompanied by simultaneous increase in plasma IgG and hemolytic complement activity. This suggests a transient shift in the balance between cell-mediated and humoral (T(H)1/T(H)2) immunity rather than immunosuppression. These results do not imply that dogs should not receive live vaccines, as the response to vaccines just seems to create a state of altered homeostasis when immunization elicits protection by humoral and cell-mediated immunity. However, these recognized compromises of immune function should be considered and vaccines still be applied only in healthy animals and strictly according to the rules and regulations given by the manufacturer. PMID- 12909409 TI - Immunohistochemical detection of SWC3, CD2, CD3, CD4 and CD8 antigens in paraformaldehyde fixed and paraffin embedded porcine lymphoid tissue. AB - Identification of the different cell types of the immune system is important for in situ studies on the pathogenesis of infectious diseases in various animals, including the pig. Unfortunately, many monoclonal anti-leukocyte antibodies are only useful for staining frozen tissue sections with inherent poor tissue morphology, and are not readily adapted to formaldehyde fixed and paraffin embedded tissue with well preserved morphology. Seven well characterised monoclonal antibodies against porcine leukocyte antigens were tested on neutral buffered paraformaldehyde fixed and paraffin embedded porcine tissue sections using the highly sensitive tyramide signal amplification system. Combining this method with different antigen retrieval techniques enabled us to detect CD2, CD3, CD4, CD8 and SWC3 antigen expressing cells in porcine lymphoid tissue. Thus, we describe herein methods for the detection of several major cell types of the porcine immune system in fixed tissue with optimal preservation of histological details. PMID- 12909410 TI - 16alpha-Bromo-epiandrosterone therapy modulates experimental feline immunodeficiency virus viremia: initial enhancement leading to long-term suppression. AB - 16alpha-Bromo-epiandrosterone (epiBr), a synthetic derivative of the natural hormone dehyroepiandrosterone (DHEA), was evaluated for its effects on feline immunodeficiency virus (FIV) infection in experimental cats. The rationale for this study was based on the ability of DHEA to significantly reduce the mortality to viral infections in mice. DHEA and epiBr also have demonstrable in vitro anti viral activity for both HIV-1 and FIV. Preliminary pharmacokinetic studies in cats demonstrated that subcutaneously injected epiBr was rapidly absorbed, completely metabolized, and nontoxic. Metabolites were excreted in both urine and feces, with the latter having the most complex pattern of breakdown products. Cats were then divided into four groups; two groups were infected with FIV and two uninfected. Two groups, one infected and one uninfected were treated on 5 consecutive days of weeks 0, 4, 8, 12 and 16 with epiBr. The remaining two groups were mock treated with the drug vehicle alone. Treatment started 1 week prior to infection and extended for 4 weeks after infection. Cats were observed for 20 weeks post-FIV infection. Infected cats had identical decreases in blood neutrophil and lymphocyte counts following, regardless of whether they were treated with epiBr or vehicle alone. The CD4/CD8 T-cell ratio was decreased following FIV exposure, but was significantly more decreased for the epiBr treated animals from week 2 post-infection onward. CD4+ T cells were decreased in FIV-infected cats treated with epiBr compared to their untreated cohort, while CD8+ T cells tended to be higher in treated animals. FIV infected cats that were treated with epiBr had over one-log higher virus loads at week 2 post-infection than non-epiBr treated cohorts. In spite of this enhanced initial viremia, the subsequent levels of virus in the blood were significantly lower in epiBr treated versus untreated animals. EpiBr treated cats had significantly higher FIV-p24 antibody responses than control cats receiving vehicle alone, although primary and secondary antibody responses to a T-cell dependent non-FIV antigen, keyhole limpet hemocyanin (KLH), were unaffected. EpiBr treatment significantly decreased the expected FIV-induced suppression of IL-12 p40 mRNA levels in peripheral blood mononuclear cells (PBMCs) observed at weeks 4, 5, 8, 9 and 16 post-infection, but had no influence on FIV-induced changes in IL-4, IL-6, IL-10, IFN-gamma, MIP 1alpha and RANTES. PMID- 12909411 TI - In vitro studies on the infection and replication of porcine circovirus type 2 in cells of the porcine immune system. AB - Porcine circovirus type 2 (PCV2) nucleic acid and/or antigens are consistently observed in cells of monocytic morphology in lesions of pigs affected by post weaning multisystemic wasting syndrome (PMWS). In this study, PCV2 antigen was detected in the cytoplasm of monocytes, pulmonary macrophages (PMs) and monocyte derived macrophages exposed to the virus in vitro, by immunofluorescence analysis (IFA) and the phenotype of these cells confirmed by detection of monocytic cell surface markers using flow cytometry. Viral antigen was not observed in lymphocytic cells. Replication of the virus in PMs was investigated further by comparison to that observed in the continuous pig kidney cell line (PK15A) using quantitative virus titration, quantitative PCR and by the detection of double stranded DNA intermediates of viral replication by Southern blotting analyses. Although increases in viral DNA and levels of infectious virus progeny and the presence of replicative intermediates, indicative of viral replication, were observed in PK15A cells, no such changes were observed in PMs in spite of the fact that infectious virus, viral antigen and viral DNA persisted in the cells for at least the duration of the experiment. These results suggest that in vivo, monocytic cells may not represent the primary target for PCV2 replication. PMID- 12909412 TI - Modulation of leukocyte populations and immune responses in sheep experimentally infected with Anaplasma (formerly Ehrlichia) phagocytophilum. AB - Anaplasma phagocytophilum infection in sheep is characterized by an immune suppression as indicated by impaired antibody response, reduced lymphocyte response and reduced oxidative burst. The effect of A. phagocytophilum infection on leucocyte populations, especially lymphocytes, was therefore investigated in six sheep experimentally infected with A. phagocytophilum, and compared with leucocyte populations from control animals.To investigate the ability of the infection to interfere with the cellular and humoral responses to specific antigens, the animals were vaccinated with commercial vaccines at the time of experimental infection, and monitored for 56 days. There were reduced percentages of gammadelta T-cells and CD4+ T-cells in peripheral blood of infected animals throughout the study period, and these cell populations showed a down-regulation of CD25 expression; while there was a relative increase in CD8+ T-cells. The reduction in CD25+ gammadelta T-cells involved a subpopulation of WC1+ gammadelta T-cells. During the first 2 weeks of the study there were reduced percentages of B-cells and leukocytes expressing MHC II and CD11b, though this decrease changed to a relative increase later in the study. The relative reductions in leucocyte populations corresponded with the observed leucopenia during the first 3 weeks post-infection, which involved lymphocyte, neutrophil and eosinophil subsets [Vet. Immunol. Immunopathol. 86 (2002) 183]. There was a reduced expression of CD11b and CD14 on granulocytes during the first 2 weeks of the study, which corresponded with the previously reported leucopenia involving neutrophils and eosinophils. Antibody responses to vaccines, lymphocyte in vitro proliferative responses to antigens and mitogens, and in vitro IFN-gamma responses to antigens were reduced up to 4 weeks after infection. PMID- 12909413 TI - Comparison of flow cytometry and image morphometry in the quantitative analysis of cell population markers in the lymph node of sheep. AB - Two approaches to the quantitative analysis of cell population markers in tissues are flow cytometry and image morphometry. To compare these methods, sheep lymph nodes were collected and analysed for CD8+ and CD21+ cell populations, which were selected to represent dispersed and concentrated cell populations, respectively. These two populations were measured as a percentage of total cell count (flow) or total tissue area (morphometry). The two populations were also measured as a percentage of respective base populations (CD2+ cells for CD8 and MHC II+ cells for CD21). A simple linear regression analysis showed that when the cell populations were assessed as a percentage of total cell count or total area, measurements obtained with flow and morphometry only correlated significantly with the dispersed CD8+ population and not with the highly concentrated CD21+ population. However, when the cell populations were assessed as a percentage of their base population, measurements obtained with flow and morphometry showed a significant correlation for both the dispersed and concentrated cell populations. This study demonstrates that measurements of lymph node cell populations obtained with the two methods are comparable, but that tissue distribution of cell populations should be considered, when the unit of measurement is chosen. PMID- 12909414 TI - Inhibition of matrix metalloproteinases ameliorates blood-brain barrier disruption and neuropathological lesions caused by avirulent Semliki Forest virus infection. AB - Semliki Forest virus (SFV) infection of mice is a useful model of viral neuropathogenesis in animals and avirulent strains such as SFV-A7 induce immune mediated demyelination and death of neurones by necrosis and apoptosis. Matrix metalloproteinases (MMPs) have been implicated in various diseases including arthritis and cancer in many species. In this report, we show that MMP-2 and MMP 9 expression is induced in the brains of mice infected i.n. with SFV-A7. Treatment of mice with the pan MMP inhibitor GM6001 ameliorated the development of SFV-induced neuropathological lesions via an effect on the integrity of the blood-brain barrier. Low levels of neuronal necrosis and demyelination in GM6001 treated mice correlated with localisation of fibrinogen staining to thin-walled blood vessels and less intense staining of the perivascular neuropil. PMID- 12909415 TI - Health impact assessment--a science and art for public health. PMID- 12909416 TI - 'Fit for purpose' health impact assessment: a realistic way forward. AB - The UK government is committed to health impact assessment (HIA) as a means of ensuring that health will be a key consideration in policy formulation and other public decision making. However there has been some debate about whether current HIA practice can reliably inform decision making. In particular consultation with stakeholders and literature reviewing, key tools used in HIA, are said to suffer from a number of conceptual and methodological problems, which can undermine the validity of the assessment. In this paper, we argue that the philosophical nature of HIA, its purpose and its contribution to the promotion of public health is still being established. We outline our own HIA practice, which is based on a broad philosophy of 'fit for purpose' i.e. what is this HIA for and what is its spatial, temporal, social and political context. We suggest that it is important to guard against unrealistic expectations and illusions of total objectivity and precision in the HIA process. HIA 'screening' is capable of delivering benefits by making policies, programmes and projects, more health conscious. Once we move beyond this basic expectation and wish to be able to make judgements about the relative health benefits of alternative courses of action, the potential resource intensiveness of the process increases considerably. Even at a high level of resource usage any conclusions reached through the HIA process will always be, in part, subjective and therefore likely to be contested. We must decide what we want, what we are prepared to legislate for and what we are prepared to pay for in the HIA process. PMID- 12909417 TI - How do we make health impact assessment fit for purpose? AB - Progress has been made in recent years in the process of health impact assessment (HIA), including community involvement. The technical side is less well developed. A minimum requirement is that there should be some consistency or robustness, so that the outcome of an HIA does not depend just on who happens to carry it out, that it is not easily swayed by the vested interests that typically surround any project, and that it can withstand legal challenge. Validity is an important criterion, as well as repeatability, as the latter can be achieved merely by propagating errors. All types of evidence should be considered legitimate, including qualitative and quantitative methods. The quality of evidence, and its generalisability, need to be carefully assessed; we should leave behind the divisive discourse around "positivism". Typically there is less information on the links from interventions (policies or projects) to changes in determinants of health than there is on the immediate precursors of health and ill-health. A practical question is, how the best existing knowledge can be made available to HIA practitioners. Other issues are more tractable than is often thought, e.g. that an HIA has to be able to trade off positive and negative impacts to different groups of people, and that the complexity of social causation prevents clear analysis of cause and effect. PMID- 12909418 TI - Migration and health impact assessment. AB - Government policies, programmes and projects can have a significant impact on health. Health impact assessments (HIAs) seek to estimate this impact, but they often do so by measuring intermediate or proxy indicators and factors that act to determine health. These measures frequently assume a static population. However, regeneration policies can work hard for several years to no apparent effect. One explanation could be migration. Families who have benefited move from the area and other, perhaps more deprived, families move in. Conversely, healthy, prosperous families may move into an improved area, giving the impression that the health of the population has changed, when in fact it is the actual population that has changed. Census data in England and Wales show that a positive correlation exists between migration within wards and deprivation scores. This paper explores the possible implications of migration for HIA. The census, NHS central register, electoral register, labour force survey, central index of the Department of Social Security, council tax database and other data sources are examined to identify what migration data are available at a local level. Factors that determine rates of migration at a local level have been reviewed, with special reference to the differences between population subgroups. The paper concludes with recommendations to take account of residential mobility and changes in migration patterns when carrying out HIAs. PMID- 12909419 TI - Hostile Hartlepool and genteel Guildford: hostility and health in 10 English towns. AB - Research from the USA suggests that cities with high mortality rates have high levels of hostility. Our aim was to replicate this research in English towns. A telephone questionnaire, based on the Cook-Medley hostility scale, was administered to random samples of adults in 10 English towns: five with high standardized mortality ratios (SMR) and five with low SMRs. The point estimate for the age-sex-weighted mean hostility score of high SMR towns was higher than that of low SMR towns (mean difference 0.2). However, the 95% confidence interval on the estimate included no difference between the two groups (-0.3-0.8). Our study does not confirm beyond doubt the findings of earlier research in the USA. PMID- 12909420 TI - Partnerships with health and private voluntary organizations: what are the issues for health authorities and boards? AB - BACKGROUND: The number of voluntary organizations active in health care is considerable. There have been recent calls for a new closer working relationship between voluntary bodies and the National Health Service. The relationship between the two healthcare sectors needs to be efficient and harmonious in the interests of patient care; however, little is known about the nature and problems in the current relationship. The present study was undertaken to examine aspects of this relationship from the point of view of health board personnel. OBJECTIVE: To identify the practices and views of Scottish health board staff concerning the funding, role and responsibility of voluntary organizations in the health sector. METHODS: A qualitative study based on in-depth interviews with health board officials in all 15 Scottish health boards. RESULTS: Policies for financial and other relationships with the voluntary sector were often not explicit. The levels and method of funding voluntary health organizations varied across boards, as did the tenure of awards (from 1 to 3 years). Demand for funding far exceeded monies available. Some health boards ensured accountability through audited accounts, annual reports and site visits; however, others thought this inappropriate for small organizations. Health boards recognized the problems of the precariousness of funding and the administrative burden of the monitoring process and the ritual of applying for funding. CONCLUSION: The uncertainties of long-term funding may impede the contribution of voluntary organizations. There is a tension between the requirements of clinical governance and the ability of small voluntary organizations to provide the necessary documentation. One proposed solution, to reduce the number of organizations, might not appeal to the voluntary sector. Future initiatives could address the problem of tailoring funding and accounting to the resources of voluntary organizations. PMID- 12909421 TI - Meta-analysis of published studies or meta-analysis of individual data? Caesarean section in HIV-positive women as a study case. AB - Very few comparative evaluations of meta-analysis of published data and meta analysis using individual patient data have appeared in the medical literature. The association between type of delivery and HIV perinatal transmission appears to be an excellent study case, given the recent publication of the meta-analysis of individual patient data by the International Perinatal HIV Group. In this paper, we report the results of a meta-analysis of the published studies, which show a statistically significant reduction of perinatal HIV transmission rate with Caesarean section. The results are surprisingly similar to those of the recently published meta-analysis of individual patient data, indicating that, in the absence of significant confounding, the two meta-analytic methods are likely to give consistent results. In this era of constrained resources for biomedical research, caution should be taken in abandoning meta-analysis of published data for studying epidemiological associations of public health interest. PMID- 12909422 TI - Caffeine consumption in drivers of heavy vehicles in Turkey. AB - Long-distance drivers in Turkey are forced to drive continuously for 15-20 h on intercity roads without sleep or assistant drivers in order that their cargo, comprising perishable goods (e.g. vegetables, fish), can be transported to their destination on time. During long journeys, drivers try to suppress their need for sleep by using substances containing caffeine as they do not want to stop for a rest. The aim of this study was to evaluate the use of gripin by long-distance drivers and its adverse effects. This cross-sectional study was conducted at the resting facilities of the Kirikkale-Ankara highway in December 1999.Sixty-five percent of long-distance drivers took gripin regularly while using their vehicles. Of these, 53.8% used gripin due to headache and 46.2% to prevent sleep. Sixty-five percent of drivers were smokers and 76.9% of smokers took gripin. It was observed that long-distance drivers tend to use substances containing caffeine in order to prevent sleep and headache. PMID- 12909423 TI - Hospital workers' opinions on the predisposing factors to blood-related work accidents in Central Hospital, Benin City, Edo State, Nigeria. AB - INTRODUCTION: Hospital workers are prone to accidental injuries during their day to-day activities at work. These accidental injuries are of importance because many of these predispose the workers to blood-borne infections. METHODS: A total of 242 hospital workers at Central Hospital, Benin City, Edo State participated in a cross-sectional study carried out between June and August 1999 aimed at finding out the predisposing factors to blood-related work accidents. The study group consisted of 53 doctors, 34 laboratory workers and 155 nurses. All doctors and laboratory workers at Central Hospital were involved in the study, while 50% of the nursing population were involved in the study. The nurses were selected using a stratified random sampling method. The respondents completed a pretested self-administered questionnaire. RESULTS: The results showed that the four main factors responsible for accidents among the hospital workers were: non availability of protective measures; desire to hasten the procedure; type of procedure; and use of inadequate/inappropriate equipment. The major factor responsible for accidents at work among hospital workers was non-availability of protective measures. RECOMMENDATIONS: It is therefore recommended that adequate materials and equipment should be provided. This study could be further validated using a qualitative study design (focus group discussions). PMID- 12909424 TI - Betel nut chewing and related factors in adolescent students in Taiwan. AB - OBJECTIVE: To investigate the prevalence of betel nut chewing and related factors among adolescent students in general, manufacturing, commercial, agricultural and medical technology schools in Taiwan. To make suggestions regarding how health authorities can reduce the number of students taking up the habit. MATERIALS AND METHODS: A cross-sectional survey was designed, and 10,288 students answered the questionnaire. RESULTS: Significantly more male than female students were betel nut chewers. The less urbanized areas were also associated with a higher prevalence of the habit. The prevalence of the habit grew with increasing age. By analysing betel nut chewing behaviour, it was found that most students started chewing betel nuts because of curiosity, and most obtained betel nuts from their friends and classmates. Betel nut chewing behaviour is definitely related to peer pressure. Most betel nut chewers did not realize that withdrawal could lead to physiological reactions, and they were ignorant of any risk to health even though they were compulsive betel nut chewers. The health-harming behaviours of both smoking and drinking were found in many betel nut chewers, particularly smoking. CONCLUSION: This study confirmed the need for a health education programme dealing with the health risks of betel nut chewing for students at risk. The health authorities could also develop a programme for betel nut chewing among Taiwan's adolescent students, emphasizing the dangers of this habit. PMID- 12909425 TI - Presence of H. pylori in the oral cavity of betel-quid ('Paan') chewers with dyspepsia: relationship with periodontal health. PMID- 12909426 TI - Communication of prenatal screening and diagnosis results to primary-care health professionals. AB - OBJECTIVE: To describe how prenatal screening and diagnostic test results are communicated to primary care health professionals. DESIGN: Postal questionnaire. SETTING: Primary care practices across London, East Anglia, the Southeast and Southwest of England. PARTICIPANTS: 714 community-based health professionals (GPs, midwives and health visitors) identified by 247 women (participants in a study of experiences after a positive prenatal diagnosis) as members of their primary care team. MAIN OUTCOME MEASURES: Description of how information about prenatal screening and diagnostic test results is communicated to health professionals working in the primary care team. Participants' views about the timeliness and sufficiency of the information they are given. RESULTS: 131 GPs, 143 midwives and 151 health visitors returned questionnaires, an overall response rate of 60.4%. Sixteen percent of primary care professionals reported that they were not usually informed of high-risk results after serum screening for Down's syndrome. Twenty eight percent were either not usually told about the possibility of abnormalities detected on ultrasound or only heard of such abnormalities from the woman herself. Even when informed, 28% of midwives and GPs did not know what action they were expected to take. Information about confirmed abnormalities is not always communicated to primary care. Only 29% were told soon enough about diagnosed abnormalities and only 17% of health professionals reported that the information they were given was always sufficient to allow them to discuss issues with their patients. The presence of specialist screening coordinators was perceived to be helpful by 76% of health professionals. Improvements in how they were told about abnormalities were suggested and these included the need for systems and protocols that ensured that quality information was communicated quickly. CONCLUSIONS: The frequent absence of means of effective communication between primary and secondary care about diagnosed abnormalities, and health professionals' uncertainties about their role has implications for the quality of care given to women who receive positive results after prenatal screening or diagnosis. There is a need for standards to be established concerning the communication of information to the health professionals responsible for the community-based care of pregnant women after a positive test result. PMID- 12909427 TI - Measuring the burden of premature death due to smoking in Korea from 1990 to 1999. AB - This study estimated the burden of premature death due to smoking in Korea between 1990 and 1999 using the years of life lost (YLL) due to premature death method. To implement this study, age-group-specific YLL due to premature death were calculated by employing the standard expected years of life lost method. YLL due to smoking were calculated based on assumptions and methods developed by the Global Burden of Disease Study Group. The burden of premature death due to smoking was estimated by multiplying the population attributable risk by the YLL of smoking-related diseases. In 1999, the burden of premature death due to smoking was 57.7% in males and 11.4% in females in Korea. The burden of premature death due to smoking increased from 1643 person years per 100,000 in 1990 to 1888 person years in 1999 for males, and increased from 151 person years in 1990 to 225 person years in 1999 for females in Korea. Our results suggest that the method employed in this study, generated in quantified terms, enabled the burden of premature death due to smoking to be obtained comparably with methods used by other international studies in this field, and thus can provide a rational basis for national health policy planning regarding premature death from smoking and the related risk factors in Korea. PMID- 12909428 TI - Education for health: case studies of two multidisciplinary MPH/MSc public health programmes in the UK. AB - Amidst the winds of change that are blowing across the UK public health (PH) landscape in relation to the essential abilities and national standards that are required for the 'art and science' of PH, the preparation for a new cadre of 'PH professionals' is already underway. Several postgraduate masters programmes in public health (MPH) have taken on board the challenge of addressing the requisite sets of skills and expertise as a guide to their content and delivery. Although there are recommendations regarding teaching PH to undergraduate medical students, little consensus seems to exist on teaching postgraduate PH to non medically qualified professionals, health managers and administrators. Employing a case study approach, this article analyses the methods used, philosophies and processes, structure and organization, outcomes to date, and lessons learnt from MPH programmes implemented at two institutions in the UK. The programmes have been initiated recently, and have had the opportunity to take on board the recent national guidelines about training standards. The findings indicate that preparatory work of the programmes, and the challenges and strengths in meeting the recent policy developments in PH training are pertinent points. The MPH programmes highlight key issues in interprofessional education and its purpose, its process and its outcomes in relation to multidisciplinary specialist practice. These programmes provide a variety of models for others wishing to develop or restructure their postgraduate PH teaching programmes. The finalization of the national standards for specialist practice in PH in the UK is encouraged, along with clearer working definitions of the domains of expertise required. Collectively, attention to these measures can ensure that the processes which teaching programmes embrace to refine their content and delivery will equip tomorrow's professionals with PH knowledge and skills. PMID- 12909433 TI - A pain neuromatrix approach to patients with chronic pain. AB - This paper presents an approach to rehabilitation of pain patients. The fundamental principles of the approach are (i) pain is an output of the brain that is produced whenever the brain concludes that body tissue is in danger and action is required, and (ii) pain is a multisystem output that is produced when an individual-specific cortical pain neuromatrix is activated. When pain becomes chronic, the efficacy of the pain neuromatrix is strengthened via nociceptive and non-nociceptive mechanisms, which means that less input, both nociceptive and non nociceptive, is required to produce pain. The clinical approach focuses on decreasing all inputs that imply that body tissue is in danger and then on activating components of the pain neuromatrix without activating its output. Rehabilitation progresses to increase exposure to threatening input across sensory and non-sensory domains. PMID- 12909434 TI - The efficacy of stretching for prevention of exercise-related injury: a systematic review of the literature. AB - The objective of this study was to conduct a systematic analysis of the literature to assess the efficacy of stretching for prevention of exercise related injury. Randomized clinical trials (RCTs) and controlled clinical trials (CCTs) investigating stretching as an injury prevention measure were selected. A computer-aided search of the literature was conducted for relevant articles, followed by assessment of the methods of the studies. The main outcome measures were scores for methodological quality based on four main categories (study population, interventions, measurement of effect, and data presentation and analysis) and main conclusions of authors with regard to stretching. One RCT (25%) and three CCTs (100%) concluded that stretching reduced the incidence of exercise-related injury. Three RCTs (75%) concluded that stretching did not reduce the incidence of exercise-related injury. Only two studies scored more than 50 points (maximum score=100 points) indicating that most of the studies selected were of poor quality. Neither of the two highest scoring RCTs showed positive effects for stretching. Due to the paucity, heterogeneity and poor quality of the available studies no definitive conclusions can be drawn as to the value of stretching for reducing the risk of exercise-related injury. PMID- 12909435 TI - Psychometric properties of a generic health measure in Chinese patients with low back pain in Hong Kong. AB - In Hong Kong, the measurement of perceived health status in patients with low back pain (LBP) can be facilitated by the availability of a health profile specifically designed for the Chinese culture. This prospective observational study investigated the psychometric properties of the generic Current Perceived Health 42 (CPH42) Profile in four separate samples (totalling 473) of Chinese patients with LBP in Hong Kong. The patients completed the CPH42 Profile and the Roland LBP Disability Scale at various points in the course of physiotherapy. Their pain intensity was measured using the 11-point pain numerical rating scale (NRS). The test-retest reliability and internal consistency of the CPH42 Profile demonstrated high intra-class correlation coefficient of 0.92 and Cronbach's alpha of 0.90. Validity was confirmed by a moderate correlation with the Chinese adaptations of the Roland LBP Disability Scale and the NRS at the commencement of physiotherapy (Spearman's correlation coefficients were 0.48 and 0.42, respectively). The responsiveness, measured from the commencement of physiotherapy to weeks 3 and 6 (standard response means of 0.33 and 0.58, respectively), were commensurate with the respective changes in pain intensity. The psychometric properties of the CPH42 Profile suggest its suitability for use as an outcome instrument in future efficacy studies on LBP intervention. PMID- 12909436 TI - Lumbar multifidus muscle size does not differ whether ultrasound imaging is performed in prone or side lying. AB - This methodological study examined the influence of body position when measuring multifidus muscle size using real-time ultrasound imaging. Previous studies examined multifidus with the subject prone but people with certain conditions cannot be studied in this position, so side lying was investigated as an alternative posture. In 20 normal females (aged 19-45 years), the cross-sectional area (CSA) of lumbar multifidus was measured at the level of the fifth lumbar vertebra (L5) on both the right and left sides, with the subject in prone and in side lying. Multifidus CSA was highly correlated between the two positions on both the right (r=0.90) and left (r=0.91) sides. Paired t-tests found no significant differences between the measurements made in the two scanning postures (right P=0.77; left P=0.16). Bland and Altman plots showed good agreement between the two positions, with no systematic difference. These findings demonstrate that measurements of lumbar multifidus at L5 can be made in either prone or side lying and a valid comparison of the results obtained in both positions can be made. PMID- 12909437 TI - Side-to-side weight-bearing asymmetry in subjects with low back pain. AB - The purpose of this project was to determine if subjects with low back pain (LBP) exhibit greater side-to-side weight-bearing (WB) asymmetry compared to healthy control subjects without LBP. This study utilized an observational double cohort design and consisted of 35 subjects with LBP and 31 healthy control subjects. Side-to-side WB asymmetry was calculated as the average of the absolute value of the difference between the right and left lower extremity from three trials. The percentage of the average side-to-side WB asymmetry relative to the total body weight was calculated to normalize expected differences in magnitude of asymmetry based on a subject's total body weight. An 11-point numeric pain rating scale was used to represent the subject's current level of pain. Patients with LBP demonstrated significantly greater normalized side-to-side WB asymmetry than healthy control subjects (8.8% vs. 3.6%, respectively, P<0.001). In patients with LBP, higher magnitudes of side-to-side WB asymmetry were significantly associated with increased pain (r=0.39, P=0.021). In conclusion patients with LBP exhibited increased side-to-side WB asymmetry compared to healthy control subjects without LBP. This asymmetry was associated with increased levels of pain. This finding is relevant for planning future studies that will attempt to provide evidence for the construct validity of manipulation by determining if side-to-side WB asymmetry normalizes after a manipulation intervention and if this improvement is associated with improvements in pain and function. PMID- 12909438 TI - Anatomical and possible clinical relationships between the calcaneofibular ligament and peroneus brevis--a pilot study. AB - The authors have observed in their clinical practice patients presenting with chronic retromalleolar pain following lateral ankle injuries. It has been hypothesized that persistent retromalleolar pain following a supination sprain may be due to peroneus brevis (PB) tendon tears (Boruta et al. 1990). The aims of this study were to investigate whether an anatomical relationship exists between the calcaneofibular ligament (CFL) and PB, and if so, the significance of this relationship in the positions of supination sprain and talar tilt test. Seven out of eight cadaveric ankles demonstrated fibrous connecting tissue between the tendon of PB and CFL. Four of the eight ankles demonstrated PB tendon abnormalities. The presence of connecting tissue between CFL and PB suggests an anatomical basis for concomitant damage to the PB tendon with a supination sprain, thus supporting the hypothesis that there may be an anatomical basis for persistent retromalleolar pain subsequent to injury to the lateral ankle complex. PMID- 12909439 TI - Measuring range of active cervical rotation in a position of full head flexion using the 3D Fastrak measurement system: an intra-tester reliability study. AB - Most external assessments of cervical range of motion assess the upper and lower cervical regions simultaneously. This study investigated the within and between days reliability of the clinical method used to bias this movement to the upper cervical region, namely measuring rotation of the head and neck in a position of full cervical flexion. Measurements were made using the Fastrak measurement system and were conducted by one operator. Results indicated high levels of within and between days repeatability (range of ICC2,1 values: 0.85-0.95). The ranges of axial rotation to right and left, measured with the neck positioned in full flexion, were approximately 56% and 50%, respectively of total cervical rotation, which relates well to the proportional division of rotation in the upper and lower cervical regions. These results suggest that this method of measuring rotation would be appropriate for use in subject studies where movement dysfunction is present in the upper cervical region, such as those with cervicogenic headache. PMID- 12909440 TI - Post-herpetic neuralgia: possible mechanisms for pain relief with manual therapy. PMID- 12909446 TI - Comments on 'Achilles tendinopathy'. PMID- 12909448 TI - A goodbye to Charlie Janeway: Charles A. Janeway Jr (1943-2003). PMID- 12909449 TI - Deciphering the relationship between central and effector memory CD8+ T cells. PMID- 12909450 TI - A genetic basis for human gammadelta T-cell reactivity towards microbial pathogens. PMID- 12909451 TI - Langerhans-cell histiocytosis: neoplasia or unbridled inflammation? PMID- 12909453 TI - Evolutionary constraints on CD1 structure: insights from comparative genomic analysis. PMID- 12909454 TI - TCR-induced NF-kappaB activation: a crucial role for Carma1, Bcl10 and MALT1. PMID- 12909455 TI - Initiation of TCR signalling revisited. PMID- 12909456 TI - What do TCR-pMHC crystal structures teach us about MHC restriction and alloreactivity? PMID- 12909457 TI - Lymphocytes transport serotonin and dopamine: agony or ecstasy? PMID- 12909458 TI - How stress influences the immune response. PMID- 12909459 TI - Heme oxygenase-1: unleashing the protective properties of heme. AB - Heme oxygenase (HO)-1 catabolizes heme into three products: carbon monoxide (CO), biliverdin (which is rapidly converted to bilirubin) and free iron (which leads to the induction of ferritin, an iron-binding protein). HO-1 serves as a "protective" gene by virtue of the anti-inflammatory, anti-apoptotic and anti proliferative actions of one or more of these three products. Administration of CO, biliverdin, bilirubin or iron-binding compounds is protective in rodent disease models of ischemia-reperfusion injury, allograft and xenograft survival, intimal hyperplasia following balloon injury or as seen in chronic graft rejection and others. We suggest that the products of HO-1 action could be valuable therapeutic agents and speculate that HO-1 functions as a "therapeutic funnel", mediating the beneficial effects attributed to other molecules, such as interleukin-10 (IL-10), inducible nitric oxide synthase (NOS2; iNOS) and prostaglandins. This Review is the third in a series on the regulation of the immune system by metabolic pathways. PMID- 12909460 TI - Hepatitis C virus infection: when silence is deception. AB - Hepatitis C virus (HCV) uses complex and unique mechanisms to prevent, evade or subvert innate and adaptive immune responses and to establish persistent infection and chronic hepatitis. Recently developed experimental systems have significantly facilitated the analysis of HCV replication, virus-host interaction and pathogenesis of chronic hepatitis and have provided new insights into the mechanisms of HCV clearance and persistence. PMID- 12909461 TI - Transmyocardial and percutaneous myocardial revascularization: current concepts and future directions. PMID- 12909462 TI - Pathogenesis of bone loss in heart transplant candidates and recipients. AB - BACKGROUND: Heart transplantation (HTX) is associated with decreased bone mineral density and changes in bone metabolism. We conducted this study to evaluate the pathophysiology of bone metabolism in HTX candidates and recipients. METHODS: Thirty-six HTX recipients were compared with 36 HTX candidates concerning biochemical parameters of bone metabolism and bone mineral density. RESULTS: Osteocalcin, bone-specific alkaline phosphatase, cross-linked-N-telopeptide of type I collagen, estradiol, serum creatinine, and blood urea nitrogen concentrations were significantly higher, whereas the calcium-creatinine ratio, thyrotropin, thyroxine, and bone mineral density were significantly lower in HTX recipients than in HTX candidates. Compared with a control group, HTX candidates had decreased renal function and increased bone resorption, whereas HTX recipients additionally had increased alkaline phosphatase and osteocalcin levels. In HTX recipients, we found positive correlations between creatinine clearance and bone mineral density; daily and cumulative cortisone doses were not associated with bone mineral density. CONCLUSIONS: In HTX candidates, disturbances in bone metabolism with increased bone resorption may be caused partly by existing low-grade renal insufficiency, regular intake of loop diuretics, and restriction of mobility. In HTX recipients, immunosuppressive therapy-glucocorticoids and cyclosporine-seem to be responsible for changes in bone metabolism. PMID- 12909463 TI - Cytokine gene polymorphisms in heart transplantation: association of low IL-10 production genotype with Quilty effect. AB - BACKGROUND: Cytokines are important modulators of post-transplant, allogeneic immune responses. In heart transplantation, endomyocardial biopsies allow monitoring of histologic and immunologic events that occur inside the graft; their correlation with risk factors condition graft outcome. Recent reports indicate that various cytokine gene allelic polymorphisms control the number of cytokines produced and may be associated with graft outcome. METHODS: We studied 71 heart transplant recipients between December 1985 and December 2000. We used sequence-specific primers (SSP) polymerase chain reaction to study interleukin-10 (IL-10) polymorphisms at -1082 (G/A), -819 (C/T), and -592 (C/A); tumor necrosis factor alpha (TNF-alpha) at -308 (G/A) and -238 (G/A); transforming growth factor beta (TGF-beta) variants at codon 10 (C/T) and codon 25 (G/C); and interferon gamma (IFN-gamma) polymorphisms at +874 (T/A). We determined the association of allele, genotype, and haplotype frequencies with the presence of histologically proven rejection episodes (according to International Society for Heart and Lung Transplantation criteria) and the presence of Quilty lesions in endomyocardial biopsy specimens. RESULTS: We found no association between the polymorphisms studied and the frequency and severity of acute and chronic rejection episodes. However, the gene frequency of allele A at IL-10 -1082, associated with decreased IL-10 production, was increased in patients with Quilty lesions (p = 0.0027, odds ratio = 2.98). Similarly, we found more AA homozygous individuals, compared with AG heterozygous and GG homozygous individuals (p = 0.0017), among patients with Quilty effect. The ATA and ACC IL-10 haplotypes also were associated with Quilty effect (p = 0.0051). CONCLUSIONS: These results suggest that genetically controlled decreased IL-10 production predisposes to the development of Quilty lesions. The decreased negative regulatory effect of IL-10 on T cells and macrophages may result in enhanced graft infiltration. PMID- 12909464 TI - Amiodarone therapy before heart transplantation as a predictor of thyroid dysfunction after transplantation. AB - BACKGROUND: In recent years we have observed new cases of thyroid disease occurring after heart transplantation (HTx). In these patients, the presence of this disease complicates their post-transplant course and occasionally results in life-threatening thyrotoxicosis. The present study examines the incidence and risk factors of thyroid disease in these patients with special emphasis on the use of amiodarone before HTx. Recommendations for follow-up of thyroid disease in HTx patients are discussed. METHODS: A study was performed on a cohort of 380 patients who received HTx in Rotterdam between January 1, 1985 and April 1, 2001. Twenty patients (5%) developed thyroid dysfunction after HTx, 10 of whom had been treated with amiodarone before HTx. RESULTS: Cumulative survival curves showed that most patients (60%) developed thyroid dysfunction within the first year after HTx (median interval between HTx and diagnosis was 9 months). Using univariate Cox regression analysis, amiodarone was a significant predictor for the development of thyroid dysfunction (hazards ratio 5.2, 95% confidence interval 2.5 to 10.8). After adjustment for age, gender, heart disease and Quetelet index, amiodarone remained a significant predictor. CONCLUSIONS: Our study clearly shows that, despite discontinuation of amiodarone treatment at the time of the HTx procedure, patients remain at risk for developing thyroid disease. HTx recipients treated with amiodarone before HTx should therefore be monitored carefully for development of thyroid dysfunction, especially within the first post-transplant year. PMID- 12909465 TI - Retransplantation in 7,290 primary transplant patients: a 10-year multi institutional study. AB - BACKGROUND: Cardiac retransplantation is a controversial procedure due to the disparity between donor heart demand and supply. METHODS: Of 7,290 patients undergoing primary cardiac transplantation between January 1990 and December 1999 at 42 institutions contributing to the Cardiac Transplant Research Database (CTRD), 106 patients later underwent a second and 1 patient a third cardiac transplant procedure. RESULTS: The actuarial freedom from retransplantation was 99.2% and 96.8% at 1 and 10 years, respectively. Reasons for retransplantation included early graft failure (n = 34), acute cardiac rejection (n = 15), coronary allograft vasculopathy (CAV, n = 39), non-specific graft failure (n = 7), and miscellaneous (n = 10). The only risk factor associated with retransplantation was younger age, reflecting the policy of preferential retransplantation of younger patients. Survival after retransplantation was inferior to that after primary transplantation (56% and 38% at 1 and 5 years, respectively). Risk factors associated with death after retransplantation included retransplantation for acute rejection (p = 0.0005), retransplantation for early graft failure (p = 0.03), and use of a female donor (p = 0.005). Survival after retransplantation for acute rejection was poorest (32% and 8% at 1 and 5 years, respectively) followed by retransplantation for early graft failure (50% and 39% at 1 and 5 years, respectively). Survival after retransplantation for CAV has steadily improved with successive eras. CONCLUSIONS: The results of retransplantation for acute rejection and early graft failure are poor enough to suggest that this option is not advisable. However, retransplantation for CAV is currently associated with satisfactory survival and should continue to be offered to selected patients. PMID- 12909466 TI - Late rejection episodes more than 1 year after pediatric heart transplantation: risk factors and outcomes. AB - BACKGROUND: Little is known about late rejection episodes after pediatric heart transplantation. We determined the frequency of late rejection episodes (>1 year) after pediatric heart transplantation, defined risk factors for its occurrence, and evaluated outcome after late rejection. METHODS: We analyzed data from 685 pediatric recipients (<18 years at transplantation) who underwent transplantation between January 1, 1993, and December 31, 1997, at 18 centers in the Pediatric Heart Transplant Study (PHTS). Probability of freedom from late rejection was determined and risk factors for late rejection and for death after late rejection were sought using univariate and multivariate analyses. RESULTS: We followed 431 patients for >1 year (median follow-up, 32.9 months) of whom 106 (24.6%) experienced 1 or more late rejection episodes (total of 178 episodes, 27 with severe hemodynamic compromise). Probability of freedom from first late rejection was 73% at 3 years and 66% at 4 years after transplantation. Risk factors (multivariate analysis) for first late rejection were >1 episode of rejection in the first year (p = 0.009), recipient black race (p = 0.0002), and older age at transplantation (p = 0.0003). Only 4 of 325 (1.2%) children who survived beyond 1 year without late rejection died compared with 26 of 106 (24.6%) with late rejection (p < 0.0001). Nine of these 26 died within 1 month of the first late rejection episode, and 17 died subsequently: 5 of acute rejection, 3 of sudden unexplained deaths, 3 of documented coronary artery disease, and 6 of other causes. Severe hemodynamic compromise with late rejection was identified as a risk factor for death among children with 1 or more episodes of late rejection. CONCLUSIONS: Approximately 25% of pediatric recipients in the PHTS who survived beyond 1 year experienced late rejection episodes. Late rejection is associated with poor survival, especially when associated with hemodynamic compromise. Absence of late rejection episodes is associated with very low risk of death during medium-term follow-up after pediatric heart transplantation. Determining the risk factors for late rejection will help to identify a cohort of patients who may benefit from enhanced rejection surveillance and treatment. PMID- 12909468 TI - Scoring system to determine need for balloon atrial septostomy for restrictive interatrial communication in infants with hypoplastic left heart syndrome. AB - BACKGROUND: Restrictive interatrial communication (IAC) causes morbidity and mortality in infants with hypoplastic left heart syndrome awaiting cardiac transplantation. We sought to create a scoring system, based on echocardiographic and clinical findings, to serve as a guide for determining the need for balloon atrial septostomy (BAS). METHODS: We retrospectively reviewed echocardiograms of 44 infants with hypoplastic left heart syndrome. Infants were studied from the time of admission to the final end-point of transplantation, Norwood procedure, or death. Seventeen infants underwent BAS for clinical indications of oxygen saturation <80% in room air. Data collected included age at BAS, maximum velocity (V(max)), and IAC diameter throughout the clinical course. We assigned higher IAC scores to smaller IAC diameter, greater V(max) through the IAC, and lower oxygen saturation value. The minimum score was 3, and the maximum score was 9. RESULTS: Only 10% of infants with a score <6 at presentation required BAS, whereas 67% of those with scores > or =6 required BAS. Higher IAC scores at presentation were associated with earlier need for BAS (p = 0.04). CONCLUSIONS: The IAC scoring system can serve as a reliable clinical guide for identifying infants with hypoplastic left heart syndrome who are likely to require BAS for relief of critically restrictive IAC while awaiting cardiac transplantation. PMID- 12909467 TI - Non-invasive detection of coronary artery disease by dobutamine-stress echocardiography in children after heart transplantation. AB - BACKGROUND: Coronary vasculopathy is the main cause of cardiac graft failure. Because yearly coronary angiography is invasive in children, a non-invasive method for detecting graft vasculopathy is needed. The aim of this study was to test dobutamine-stress echocardiography in a pediatric population to determine its feasibility, safety and reliability in the detection of graft coronary artery disease. METHODS: Eighteen patients, aged 2 days to 16.8 years at transplantation (mean 8.4 years), underwent 44 dobutamine-stress echocardiography (DSE) exams, at a follow-up of 1.1 to 11.8 years (mean 5.1 years). Selective coronary angiography was performed for comparison. Echocardiographic recordings were obtained in 4 standard views of the left ventricle and measurements carried out within the frames of a 16-segment model. Segmental scores of contractility were obtained for each segment and a total segmental contractility index was calculated at each stage. RESULTS: All patients reached the maximum dose stage. Maximum heart rate was 57% to 90% of predicted maximum. Maximum systolic blood pressure reached 190 mmHg. Segmental scores were normal in 37 and abnormal in 7 cases. Echographic results were concordant with angiography in 82% and discordant in 18% of the cases (4 negative DSEs with minor angiographic lesions, 2 positive DSEs with normal angiography), but there was no significant angiographic lesion with normal DSE. CONCLUSIONS: DSE is a safe and highly feasible non-invasive technique in transplanted children. A normal DSE study successfully predicts the absence of significant coronary artery disease in the post-transplant population. PMID- 12909469 TI - Long-term follow-up of arrhythmias in pediatric orthotopic heart transplant recipients: incidence and correlation with rejection. AB - BACKGROUND: Arrhythmias in adult orthotopic heart transplant (OHT) recipients are common and have been used as predictors of rejection. Because of the paucity of information in pediatric OHT recipients, the purpose of this study was to determine the incidence and correlation of arrhythmias with rejection or with coronary artery disease (CAD) in children. METHODS: We retrospectively reviewed the records, electrocardiograms (ECGs), and 24-hour ambulatory ECGs of patients who underwent OHT from January 1984 to December 1999. We excluded arrhythmias occurring in the first 2 weeks after OHT. RESULTS: Sixty-nine patients underwent OHT, received triple-immunosuppression therapy, were discharged home, and have been followed for a mean of 4.7 years (0.3-13 years). Each patient had an average of 10 ECGs and three 24-hour ECGs. Twenty-six patients had 33 arrhythmias: sinus bradycardia (n = 9), atrial tachycardia (n = 9), ventricular tachycardia (n = 3), and Wenckebach periodicity (n = 6). Sinus bradycardia was treated with theophylline in 8 patients, and 2 required pacemakers. Atrial tachycardias (atrial flutter in 4 patients and atrial ectopic tachycardia in 5) were treated with digoxin, propranolol, or procainamide. Ventricular tachycardia was treated with mexiletine, lidocaine, and amiodarone. There were 65 episodes of rejection, 20 of which were moderate/severe (> or =3B). Only Wenckebach was associated with the presence of either rejection or CAD (p < 0.05). CONCLUSIONS: We noted clinically significant arrhythmias in 38% of the pediatric OHT recipients. Sinus bradycardia, atrial tachyarrhythmias, and ventricular tachycardia occurred with the same frequency. Only new-onset Wenckebach periodicity was noted in the presence of either CAD or rejection. No arrhythmia was of negative predictive value for rejection or CAD. From this data, we suggest that new-onset Wenckebach prompt evaluation for rejection or CAD. PMID- 12909470 TI - Weight gain after lung transplantation. AB - BACKGROUND: Weight gain is frequently observed after lung transplantation, but the magnitude, predictors and implications of weight gain after lung transplant are unknown. METHODS: This retrospective cohort study included 826 lung transplant recipients randomly selected from 12 international transplant centers. We included adult patients with available weight data at baseline and 1 year post transplant. We examined demographic and clinical predictors of first year weight gain using a multiple linear regression model (n = 579) with percent weight change as the dependent variable. To study the association between first year weight gain and subsequent survival, we performed a Cox proportional hazards analysis. p < 0.05 was considered statistically significant. RESULTS: The median weight change was 10% (range -32% to 84%). On multi-variate analysis, increasing age and prolonged mechanical ventilation were inversely associated with weight gain; obstructive disease, interstitial disease and increasing ischemic time were positively associated with weight gain. Increasing baseline weight was negatively associated with weight gain in patients with obstructive and interstitial disease. The model accounted for 14% of the variance in weight gain. Patients with weight gain above the median had better subsequent survival (adjusted hazard ratio 0.61, 95% confidence interval 0.41 to 0.90). Infection was a more common cause of death in these patients, whereas malignant deaths were more frequent in patients with below-median weight gain. CONCLUSIONS: Substantial weight gain occurs in the first year after lung transplantation. The predictors of weight gain may be used to target high-risk patients for early intervention. Higher weight gain is associated with better subsequent survival. PMID- 12909471 TI - Safety of inhaled nitric oxide after lung transplantation. AB - BACKGROUND: The present study tests the hypothesis that therapy with inhaled nitric oxide (iNO) at the time of lung transplantation in patients undergoing bilateral angle lung transplantation: (i) is safe; and (ii) does not increase either the duration of mechanical ventilation or the incidence of acute graft dysfunction. METHODS: We conducted a prospective, non-randomized trial of iNO at 20 parts per million. The treatment group was comprised of 14 patients (10 females, 4 males) undergoing lung transplantation to address severe end-stage lung disease and pulmonary hypertension (mean pulmonary artery pressure > 30 mmHg). Clinical and histologic parameters were compared with 22 historical control subjects who were matched with the study population for age, diagnosis and disease severity (17 females, 5 males) and had undergone lung transplantation in the preceding 2-year time period. No significant differences were noted between the 2 study groups at baseline. RESULTS: No toxic effect of iNO treatment was evident. Although the incidence of acute graft dysfunction was the same in both groups, the occurrence of acute graft rejection in the initial 4 weeks after transplant was less frequent in the iNO group than in the control group (7% vs 32%, p = 0.05). Fifty percent of the treatment group, as compared with 22% of the control group, were discharged from the hospital within 2 weeks of the procedure (p = 0.05). CONCLUSIONS: Early initiation of iNO in lung transplant patients with pulmonary hypertension is safe and may decrease the incidence of acute graft rejection. We speculate that iNO may exert an immunomodulatory effect. PMID- 12909472 TI - Effect of switching from cyclosporine to tacrolimus on exhaled nitric oxide and pulmonary function in patients with chronic rejection after lung transplantation. AB - BACKGROUND: Previous studies have demonstrated that shifting immunosuppressive therapy from cyclosporine (CyA) to tacrolimus (FK) may arrest the decline in forced expiratory volume in 1 second (FEV(1)) during chronic rejection after lung transplantation. Exhaled nitric oxide (eNO) has been shown to be elevated during chronic rejection. We report the concomitant stabilization of FEV(1) and decrease in eNO after changing from CyA to FK therapy in patients with chronic rejection after lung transplantation. METHODS: We used a prospective design. The study included 10 lung transplant patients (5 men and 5 women), mean age 44 +/- 14 years at time of transplantation, with a progressive decline in FEV(1) that was attributed to chronic rejection. Four patients underwent heart-lung transplantation and 3 had a sequential single and 3 a single-lung transplantation. The switch from CyA to FK occurred at 36 +/- 23 months after transplantation (Time 0). The eNO was measured using a chemiluminescence analyzer, according to standardized European Respiratory Society (ERS) criteria. RESULTS: At Time 0, there were 6 patients in bronchiolitis obliterans syndrome (BOS) Stage 0-p, with a mean decline in FEV(1) of 15 +/- 3%; 2 in BOS Stage 1; and 2 in BOS Stage 2. Compared with the best post-operative FEV(1), there was a progressive and significant decline until Time 0, from 2.56 +/- 0.9 liters to 2.03 +/- 0.94 liters (p = 0.0047). Thereafter, FEV(1) stabilized: 2.03 +/- 0.94 liters at Time 0 and 2.05 +/- 0.94 liters 6 months later (p = non-significant). Concomitantly, there was a gradual increase in eNO during the 6 months before Time 0, from 11.4 +/- 2.5 ppb at the time of best FEV(1) to 20.5 +/- 14.8 ppb at Time 0. After switching, there was a non-significant decline in eNO, from 20.5 +/ 14.8 ppb to 14.9 +/- 5.4 ppb. There was no significant difference in eNO levels between the patients in BOS Stage 0-p and patients in higher BOS stages at either timepoint in the study. CONCLUSIONS: This study illustrates that a switch from CyA to FK can stabilize pulmonary function in lung transplant patients with chronic rejection. This stabilization of FEV(1) is accompanied by a decrease in eNO, indicating that this treatment shift can reduce inflammation of airways during the course of chronic rejection. Consequently, measuring eNO may be extremely valuable in guiding the treatment of chronic rejection after lung transplantation. PMID- 12909473 TI - LVAD bloodstream infections: therapeutic rationale for transplantation after LVAD infection. AB - INTRODUCTION: Patients who have ventricular assist devices (VADs) and experience bloodstream infection (BSI) have high mortality. We addressed 2 questions raised by the United Network for Organ Sharing (UNOS) priority policy for this problem: 1) Are organs wasted on this ultra-high-risk group? 2) Can device-related BSI be differentiated from transient BSI? METHODS: Patients with VADs who underwent heart transplantation from 1987 to 2001, who had BSI during VAD support, and who had positive cultures at VAD explant (device-related BSI, n = 10) were compared with those with negative cultures at explant (non-device-related BSI, n = 11). RESULTS: Patients with device-related BSI had an 80% (8/10) rate of persistent bacteremia; 30 days and 1 year after transplantation, mortality was 14% and 26%, respectively. Non-device-related BSI (n = 11) persisted in 18% (2/11); peri operative and 1-year mortalities were 9% and 13%. Duration of VAD support predicted infection (132 vs 48 days, p < 0.001); hypo-albuminemia (2.9 +/- 0.5 mg/dl vs 3.3 +/- 0.8 mg/dl, p < 0.05), and a resistant organism predicted a device-related BSI. These patients had increased intubation requirements and had increased creatinine concentration during the first post-operative week, with no difference in liver function, blood loss, transfusions (packed red blood cells, fresh frozen plasma, or platelets), or hemodynamic stability vs patients with non device BSI. Despite decreased immunosuppression, we found no difference in acute rejection events with device-related BSI. Re-infection with the pre-operative organism occurred in only 1 patient per group. CONCLUSIONS: These data suggest that urgent (Status 1A) cardiac transplantation is effective in stable patients with device-related BSI, and these data support the current UNOS policy. However, an extra-device source of BSI should be excluded by considering the isolated organism, the baseline nutritional status, and other risk factors. PMID- 12909474 TI - Functional evidence of reversible ischemic injury immediately after the sympathetic storm associated with experimental brain death. AB - BACKGROUND: Acute brain death from increased intracranial pressure results in a transient increase in myocardial adenosine and lactate, which indicates that oxygen demand exceeds oxygen delivery during the sympathetic "storm". The aim of this study was to determine the functional significance of this period of ischemia. METHODS: Brain death was inflicted on 40 Westran pigs (36.5-68.0 kg) by inflating a 21-ml subdural balloon over 3 minutes. In 38 animals, micromanometry and sonomicrometry were used to obtain left ventricular pressure-volume loops to determine the preload recruitable stroke work (PRSW) relationship. Data files were recorded before and at 15-minute intervals after beginning balloon inflation. Plasma troponin I was measured before and 60 minutes after beginning balloon inflation in the 38 instrumented and 2 non-instrumented animals. RESULTS: All animals experienced the classical sympathetic storm. The slope of the PRSW relationship decreased, and the volume-axis intercept shifted to the right 15 minutes after beginning balloon inflation (p < 0.0001). Progressive incremental recovery (leftward shift) occurred between subsequent time points (p < or = 0.0018). In the instrumented animals, the mean plasma troponin I level increased from 1.4 +/- 1.6 microg/liter to 2.8 +/- 2.3 microg/liter (p < 0.001). However, troponin I was not detected before or after induction of brain death in the plasma of either non-instrumented animal (p = 0.001). CONCLUSIONS: The sympathetic storm produced transient contractile dysfunction, consistent with ischemic injury. However, troponin I release reflected surgical instrumentation and not brain death. PMID- 12909475 TI - Cardioprotection by cariporide after prolonged hypothermic storage of the isolated working rat heart. AB - BACKGROUND: Inhibition of the sodium-hydrogen (Na(+)-H(+)) exchanger decreases the extent of ischemia-reperfusion injury in the myocardium. Inhibition may also improve preservation of hearts stored for transplantation. Our aim was to characterize the dose response and to determine optimal timing for administering cariporide, an Na(+)-H(+) exchange inhibitor, during prolonged hypothermic storage. METHODS: We used the rat isolated working-heart model to measure cardiac function. To determine the optimal dose of cariporide, hearts received either no treatment (control) or incremental doses of cariporide (1, 3.2, 10, or 30 micromol/liter) before storage and during reperfusion. Hearts were arrested with and stored in an extracellular-based cardioplegic solution at 2 to 3 degrees C for 6 hours. To determine optimal timing, we arrested a group of hearts with and stored them in a cariporide-supplemented (10 micromol/liter) cardioplegic solution but did not pre-treat them with cariporide. Finally, we treated a separate group of hearts with 10 micromol/liter cariporide before, during, and after storage. RESULTS: Recovery of cardiac function in control hearts was poor. The cardioprotective effect of cariporide was dose dependent, with maximal protection observed at a concentration of 10 micromol/liter. Storing hearts in a cariporide-supplemented cardioplegic solution did not result in better recovery of cardiac function compared with cariporide given before storage and during reperfusion. Moreover, recovery of cardiac function was significantly worse in hearts that had not been pre-treated with cariporide. CONCLUSIONS: Sodium hydrogen-exchange inhibition with cariporide significantly protects the hypothermic ischemic rat heart, increasing cardiac function after reperfusion. The timing of cariporide administration is an important determinant of this cardioprotection. PMID- 12909477 TI - Progressive multifocal leukoencephalopathy in a lung transplant recipient. AB - Progressive multifocal leukoencephalopathy (PML) is a sub-acute, demyelinating disease of the brain caused by a human polyomavirus. We describe a patient with the onset of PML 7 months after lung transplantation. The patient was treated with immunosuppressive modulation and cidofovir, a new anti-viral therapy for PML, with stabilization of the symptoms. We also review the 4 additional reports in the literature of PML after heart and lung transplantation. Progressive multifocal leukoencephalopathy may become more prevalent as the population of heart and lung transplantation recipients increases. PMID- 12909476 TI - Aortic regurgitation in the heterotopic rat heart transplant: effect on ventricular remodeling and diastolic function. AB - OBJECTIVES: Use of the heterotopic rat cardiac isograft model is limited by ventricular atrophy attributable to the left ventricle's non-working state. Previous studies indicate that increased left ventricular pressure-volume work minimizes atrophy. We used a simpler approach to increase ventricular work, imposing aortic regurgitation on the transplant. We hypothesized that this would prevent atrophy and preserve left ventricular compliance. METHODS: We analyzed heterotopic transplants with aortic valvotomy and without aortic valvotomy (controls). Recipient native hearts served as separate controls. After 15 to 25 days, we measured cardiac wet weight, dry weight, and water content of all groups and measured echocardiographic left ventricular wall thickness and end-diastolic and end-systolic diameters in both transplant groups. Left ventricular volume infusions yielded pressure-volume data that we analyzed using regression methods. RESULTS: Aortic regurgitant transplants weighed more than control transplants (dry weight, 0.109 +/- 0.013 g vs 0.097 +/- 0.016 g; p = 0.020, 2-way analysis of variance), but all transplants weighed less than native hearts weighed (p = 0.001). Control transplants were less compliant than regurgitant transplants (p = 0.002), but the latter were similar to their own native hearts (p = 0.34). Wall thickness decreased in regurgitant vs control transplants (p = 0.020, Student's t test), but end-diastolic and end-systolic diameters increased (p < or = 0.001). CONCLUSIONS: Aortic regurgitation in heterotopic transplants improves left ventricular compliance through chamber dilatation without preventing atrophy. Moderate acute aortic regurgitation affects ventricular remodeling more than it stimulates myocardial hypertrophy. Smaller end-diastolic diameter, greater wall thickness, and myocardial edema may explain decreased compliance in non-working transplants. PMID- 12909478 TI - Laparoscopic treatment of polycystic ovaries: is it time to relinquish the procedure? AB - OBJECTIVE: To evaluate the role of laparoscopic ovarian drilling and treatment with metformin in the management of the polycystic ovary syndrome (PCOS). DESIGN: A literature search was conducted using the keywords laparoscopy, laparoscopic ovarian drilling, laparoscopic ovarian diathermy, PCOS, metformin, and ovulation. The MEDLINE and EMBASE databases and the Cochrane Database of Systematic Reviews were searched. RESULTS: No randomized comparisons have been done between laparoscopic ovarian drilling and metformin therapy. However, the ovulation and pregnancy rates appear to be similar for both techniques. Both treatments decrease the incidence of ovarian hyperstimulation and the cancellation rate of IVF cycles. However, unlike laparoscopic ovarian drilling, metformin may decrease the incidence of type 2 diabetes and coronary heart disease. CONCLUSION(S): Given the similar magnitude of the results without the potential risks and complication of surgery, we propose that laparoscopic ovarian drilling should be used sparingly in favor of less invasive treatment with metformin. PMID- 12909479 TI - Androgen excess is the key element in polycystic ovary syndrome. PMID- 12909480 TI - The importance of insulin resistance in polycystic ovary syndrome. PMID- 12909481 TI - What are the key features of importance in polycystic ovary syndrome? PMID- 12909482 TI - Hyperandrogenemia is necessary but not sufficient for polycystic ovary syndrome. PMID- 12909483 TI - We should avoid the indiscriminate use of insulin sensitizers in women with polycystic ovary syndrome. PMID- 12909484 TI - Distinctive proliferative phase differences in gene expression in human myometrium and leiomyomata. AB - OBJECTIVE: To gain a comprehensive view of the gene expression and regulation involved in uterine leiomyomata and matched normal myometrium using oligonucleotide microarray-based hybridization analysis. DESIGN: Retrospective analyses of tissue obtained in a prospective randomized clinical study. SETTING: Academic institution. PATIENT(S): Seven patients with leiomyomata scheduled for surgery during the proliferative phase. INTERVENTIONS(S): Seven paired samples of leiomyomata and adjacent myometrium were obtained from patients undergoing hysterectomy. MAIN OUTCOME MEASURE(S): The total RNA extracted from leiomyomata and myometrium was used for gene expression profiling of 6800 human genes using high-density oligonucleotide microarrays. In addition, reverse transcriptase semiquantitative polymerase chain reaction and immunohistochemistry were used to validate tumor-specific gene expression. RESULT(S): A comparison of expression patterns in each paired sample revealed 68 genes significantly up- or down regulated in each paired tissue sample, of which 23 genes showed increased expression and 45 showed decreased expression in leiomyomata compared with normal myometrium. Cluster analysis supported the relevance of these candidate genes for distinguishing between normal myometrium and leiomyomata biologic activity. CONCLUSION(S): Expression profiling of uterine leiomyomata using high-density oligonucleotide microarrays yields signature patterns that reflect the distinctive differences between normal human myometrium and leiomyomata during the proliferative phase. These observations suggest that a number of genes are involved in the tumorigenesis of leiomyomata. PMID- 12909485 TI - Genome biology and gynecology: the application of oligonucleotide microarrays to leiomyomata. PMID- 12909486 TI - New and old regulators of uterine leiomyoma growth from screening with DNA arrays. PMID- 12909487 TI - Strategy for elucidating differentially expressed genes in leiomyomata identified by microarray technology. AB - OBJECTIVE: cDNA microarray technology identifies genes that are differentially expressed between tissues. Our previous study identified several genes that might contribute to the fibroid phenotype. We therefore sought to confirm genes involved in three distinct signal transduction pathways. DESIGN: Evaluation of differential mRNA and protein expression of Dlk, Frizzled-2, and CD-24 in fibroids compared with adjacent myometrium. University hospital. PATIENT(S): Five women undergoing medically indicated hysterectomy for symptomatic fibroids. INTERVENTION(S): Microarray analysis of up to 33000 genes, reverse transcriptase polymerase chain reaction (RT-PCR), real-time RT-PCR, Western blot, and immunohistochemistry. MAIN OUTCOME MEASURE(S): Expression of mRNA transcripts and protein in fibroid compared with myometrium.A more extensive microarray confirmed differential expression of Frizzled-2 and CD-24 but did not confirm Dlk overexpression. RT-PCR and real-time PCR demonstrated equivalent Dlk mRNA expression between fibroid and myometrium (ratio, 1.02), a slight Frizzled-2 overexpression (ratio, 2.09), and robust CD-24 overexpression in fibroids (ratio, 12.35). Western blot and immunohistochemistry confirmed Frizzled-2 overexpression, but did not confirm Dlk overexpression. CONCLUSION(S): Microarray technology is the first phase of tissue evaluation, but changes in gene expression must be confirmed. Confirmed genes can then be used to generate hypotheses testing their involvement in fibroid development. PMID- 12909489 TI - Microarray analysis in fibroids: which gene list is the correct list? PMID- 12909488 TI - Bench to bedside: clinical opportunities for microarray analysis. PMID- 12909490 TI - The story behind the American Society for Reproductive Medicine's prevention of infertility campaign. PMID- 12909491 TI - Gonadotropin-releasing hormone agonist treatment reduced serum interleukin-6 concentrations in patients with ovarian endometriomas. AB - OBJECTIVE: To determine whether serum interleukin (IL)-6 can be measured in patients with ovarian endometriomas and whether these measurements are useful in managing this disease. DESIGN: A controlled clinical study and an in vitro study. SETTING: Department of Obstetrics and Gynecology, Tottori University, Japan.Twenty-two patients with ovarian endometriomas. INTERVENTION(S): Laparoscopic cystectomy for ovarian endometriomas was performed. Gonadotropin releasing hormone (GnRH) agonist was administered for 3 months in nine patients before laparoscopic surgery. Endometriotic stromal cells obtained from patients with endometriomas with or without GnRH agonist treatment were cultured. MAIN OUTCOME MEASURES(S): IL-6 concentrations in serum or supernatant of the cell culture were measured using ELISA. RESULTS: The serum concentration of IL-6 in patients with endometriomas was higher at the time of diagnosis than in those without endometriomas. Laparoscopic surgery significantly reduced serum levels of IL-6. Serum IL-6 concentrations also decreased after treatment with GnRH agonist. IL-6 production was attenuated in the endometriotic stromal cells obtained from patients with GnRH agonist treatment compared with patients without such treatment. CONCLUSION(S): GnRH agonist treatment may decrease IL-6 production in endometriotic cells. Measurement of serum IL-6 concentrations may be of value in managing patients with endometriomas. PMID- 12909492 TI - Comparison of a levonorgestrel-releasing intrauterine device versus expectant management after conservative surgery for symptomatic endometriosis: a pilot study. AB - OBJECTIVE: To determine whether the frequency and severity of dysmenorrhea are reduced in women with symptomatic endometriosis in whom a levonorgestrel releasing intrauterine device (Lng-IUD) is inserted after operative laparoscopy compared with those treated with surgery only. DESIGN: Open-label, parallel group, randomized, controlled trial. SETTING: A tertiary care and referral center for patients with endometriosis. PATIENTS(S): Parous women with moderate or severe dysmenorrhea undergoing first-line operative laparoscopy for symptomatic endometriosis. INTERVENTION(S): Randomization to immediate Lng-IUD insertion or expectant management after laparoscopic treatment of endometriotic lesions. Proportions of women with recurrence of moderate or severe dysmenorrhea in the two study groups 1 year after surgery and overall degree of satisfaction with treatment. Moderate or severe dysmenorrhea recurred in 2 of 20 (10%) subjects in the postoperative Lng-IUD group and 9/20 (45%) in the surgery-only group. Thus, a medicated device inserted postoperatively will prevent the recurrence of moderate or severe dysmenorrhea in one out of three patients 1 year after surgery. A total of 15/20 (75%) women in the Lng-IUD group and 10/20 (50%) in the expectant management group were satisfied or very satisfied with the treatment received. CONCLUSION(S): Insertion of an Lng-IUD after laparoscopic surgery for symptomatic endometriosis significantly reduced the medium-term risk of recurrence of moderate or severe dysmenorrhea. PMID- 12909493 TI - Laparoscopic uterosacral ligament resection for dysmenorrhea associated with endometriosis: results of a randomized, controlled trial. AB - OBJECTIVE: To evaluate the efficacy of laparoscopic resection of the uterosacral ligaments in women with endometriosis and predominantly midline dysmenorrhea. DESIGN: Randomized controlled trial. SETTING: Two academic departments. One hundred eighty patients undergoing operative laparoscopy as first-line therapy for stage I to IV symptomatic endometriosis. INTERVENTION(S): Operative laparoscopy including uterosacral ligament resection or conservative surgery alone. MAIN OUTCOME MEASURE(S): Proportion of women with recurrence of moderate or severe dysmenorrhea 1 year after surgery. RESULT(S): No complications occurred. Among the patients who were evaluable 1 year after operative laparoscopy, 23 of 78 (29%) women who had uterosacral ligament resection and 21 of 78 (27%) women who had conservative surgery only reported recurrent dysmenorrhea. The corresponding numbers of patients at 3 years were 21 of 59 (36%) women and 18 of 57 (32%) women, respectively. Time to recurrence was similar in the two groups. Pain was substantially reduced, and patients in both groups experienced similar and significant improvements in health-related quality of life, psychiatric profile, and sexual satisfaction. Overall, 68 of 90 (75%) patients in the uterosacral ligament resection group and 67 of 90 (74%) patients in the conservative surgery group were satisfied at 1 year. CONCLUSION(S): Addition of uterosacral ligament resection to conservative laparoscopic surgery for endometriosis did not reduce the medium- or long-term frequency and severity of recurrence of dysmenorrhea. PMID- 12909494 TI - Elevated early follicular gonadotropin levels in women with unexplained infertility do not provide evidence for disordered gonadotropin-releasing hormone secretion as assessed by luteinizing hormone pulse characteristics. AB - OBJECTIVE: To determine whether women with rigorously defined unexplained infertility demonstrated altered GnRH secretion, as reflected by serum LH secretion patterns. DESIGN: Prospective observational study. SETTING: National Center for Infertility Research at Michigan. PATIENT(S): Nine women with rigorously defined unexplained infertility and 11 healthy, parous age-matched control women.Gonadotropin-releasing hormone (25 ng/kg) as a bolus injection. MAIN OUTCOME MEASURE(S): Daytime pulse patterns of LH secretion measured every 10 minutes; mean serum concentrations of LH, FSH, E(2), P, PRL, and cortisol; and response to a physiologic dose of GnRH in the early follicular, late follicular, mid-luteal, and late luteal phases of the same menstrual cycle. RESULT(S): Serum LH pulse frequency and pulse amplitude and LH secretion in response to a physiologic bolus of GnRH were not significantly different in unexplained infertility patients at any phase of the cycle. Luteinizing hormone pulse frequency and amplitude, as well as response to GnRH, varied significantly across the cycle. Mean early follicular serum LH and FSH concentrations were significantly higher in unexplained infertility patients than in fertile control subjects (LH: 5.31 +/-.51 vs. 4.03 +/-.33 [mIU/mL +/- SEM]; FSH: 5.81 +/-.63 vs. 3.80 +/-.45) but were not different at any other phase of the cycle. CONCLUSION(S): These data do not support the hypothesis that unexplained infertility is caused by an abnormality in pulsatile GnRH secretion or abnormal pituitary sensitivity to GnRH. However, the results are consistent with a difference in negative feedback from the ovary to the pituitary in unexplained infertility patients that is suggestive of diminished ovarian reserve. PMID- 12909495 TI - Follicular response and pregnancy among infertile women undergoing ovulation induction and intrauterine insemination. AB - OBJECTIVE: To determine the relationship between ovarian follicular response (number and diameter of follicles) to ovulation induction agents and pregnancy. DESIGN: Retrospective cohort study.Hospital-based center of reproductive medicine. PATIENT(S): Three hundred twenty-two clinically diagnosed infertile couples undergoing 1483 cycles of ovulation induction and IUI. Oral and injectable fertility drugs were administered for 2-10 cycles; ultrasound follicular measurements were recorded around time of ovulation; hCG was administered when follicles were 18-20 mm or more in diameter; IUI was performed 36-40 hours after administration of hCG. MAIN OUTCOME MEASURE(S): A first positive pregnancy test as indicative for conception. Women with follicular diameters >or=20 mm were less likely to become pregnant as compared to women with diameters between 15.00 and 19.99 mm (risk ratio [RR] = 0.58, 95% confidence interval [CI] = 0.35, 0.97). Women >or=30 years of age were half as likely to become pregnant as compared to women <30 years (RR = 0.51, 95% CI = 0.30, 0.85). Likelihood of pregnancy increased by 21% for each prior pregnancy (RR = 1.21, 95% CI = 1.00, 1.47). CONCLUSION(S): Maternal age, gravidity, and follicular diameters around the time of insemination are prognostic factors in the likelihood of pregnancy. PMID- 12909496 TI - Body mass index and uterine receptivity in the oocyte donation model. AB - OBJECTIVE: To evaluate the relationship of body mass index (BMI) to uterine receptivity under conditions of programmed hormonal support and standardized embryo quality. DESIGN: Retrospective cohort study.A tertiary referral center. PATIENTS: Ninety-seven consecutive first-cycle recipients of anonymous oocyte donation. After programmed hormone replacement, recipients had transfer of embryos derived from oocyte donation. Anonymous oocyte donors received ovarian stimulation and underwent transvaginal ultrasound-guided oocyte retrieval. SETTING: A receiver operator characteristic (ROC) curve of implantation versus BMI. Area under the ROC curve was 0.51, 95% confidence interval (CI) 0.41-0.62, suggesting no relationship between BMI and implantation. There was no difference in implantation rates between obese (BMI >or=30) and nonobese (BMI <30) recipients, odds ratio 1.1, 95% CI 0.5-2.4. CONCLUSION(S): Uterine receptivity was unimpaired in women with increased BMI when hormonal support and embryo quality were standardized. PMID- 12909498 TI - A prospective assessment of the predictive value of basal antral follicles in in vitro fertilization cycles. AB - OBJECTIVE: To determine the predictive value and define threshold values for basal antral follicle count in patients undergoing IVF. DESIGN: Prospective cohort analysis. Tertiary care center. Two hundred eighty-nine patients. Transvaginal ultrasonography before starting gonadotropin administration. MAIN OUTCOME MEASURES: Number of oocytes retrieved, basal hormone levels, and cycle outcomes. RESULTS: Pregnant patients had significantly more antral follicles (13.8 +/- 7.5 vs. 12.4 +/- 10.0). Patients in whom cycles were canceled had significantly fewer antral follicles (7.6 +/- 4.8 vs. 13.7 +/- 8.8). Antral follicle count significantly correlated with most prestimulation and poststimulation IVF variables. Threshold analysis demonstrated a lower pregnancy rate (23.5% vs. 57.6%) and a higher cancellation rate (41% vs. 6.4%) associated with having four or fewer antral follicles. CONCLUSION(S): The basal antral follicle count identified patients who responded poorly to IVF stimulation. Having four or fewer antral follicles was associated with a high cancellation rate (41%) and, in patients without a cancelled cycle, a low pregnancy rate (23%). However, no antral follicle count absolutely predicted pregnancy or cycle cancellation. PMID- 12909497 TI - Pronuclear morphology and chromosomal abnormalities as scoring criteria for embryo selection. AB - OBJECTIVE: To verify whether a correlation exists between pronuclear zygote morphology and the chromosomal condition of preimplantation embryos. DESIGN: Prospective analysis of pronuclear zygote morphology and preimplantation genetic diagnosis (PGD) for aneuploidy of the resulting embryos. SETTING: Reproductive medicine unit, day surgery clinic. PATIENT(S): Seventy-seven patients undergoing 107 PGD cycles because of advanced maternal age (77 cycles) or previous IVF failures (30 cycles). INTERVENTION(S): Evaluation of pronuclear zygote morphology and chromosomal condition of the resulting embryos. MAIN OUTCOME MEASURE(S): Rate of embryo development, proportion of euploid embryos, and distribution of chromosomal abnormalities. The position of pronuclei within the ooplasm, the size and distribution of nucleoli, and the orientation of polar bodies with respect to pronuclei were highly predictive for the presence of complex chromosomal abnormalities in the developing embryos; zygotes with juxtaposed pronuclei, large size nucleoli, and polar bodies with small angles subtended by pronuclei and polar bodies were the configurations associated with the highest rates of euploidy. CONCLUSION(S): The combination of the patterns related to pronuclear zygote morphology indicated four configurations where the proportion of chromosomally normal embryos was significantly higher compared with the other configurations, suggesting the validity of this scoring system for the selection of embryos generated by PGD patients. PMID- 12909499 TI - Assessing the clinical utility of in vitro fertilization with intracytoplasmic sperm injection in human immunodeficiency virus type 1 serodiscordant couples: report of 113 consecutive cycles. AB - OBJECTIVE: To assess the utility and safety of in vitro fertilization (IVF) with intracytoplasmic sperm injection (ICSI) in human immunodeficiency virus-1 (HIV-1) serodiscordant couples. DESIGN: Retrospective study. SETTING: University-based practice. PATIENT(S): HIV-1 seropositive men and seronegative women undergoing IVF-ICSI.IVF-ICSI, HIV testing of female partner and infants. MAIN OUTCOME MEASURE(S): IVF performance, pregnancy rates, and HIV-1 seroconversion rate. RESULT(S): We initiated 113 IVF cycles in 61 serodiscordant couples. Due to poor ovarian response, 11.5% of cycles were canceled. The number of oocytes collected per retrieval was 17.11 +/- 9.52 (2 to 47), yielding 13.90 +/- 8.12 (1 to 42) mature oocytes suitable for ICSI, and 9.34 +/- 5.45 (0 to 24) normal fertilized oocytes. We transferred 3.54 +/- 1.09 (1 to 6) embryos. The overall clinical pregnancy rate was 44.8% per embryo transfer (ET); ongoing/delivered pregnancy rate was 36.5% per ET, with a 57.1% multiple gestation rate. Cumulatively, 50.8% of couples achieved a successful pregnancy through IVF-ICSI, 54.1% when including frozen ET cycles. There were no HIV-1 seroconversions in patients or delivered babies. CONCLUSION(S): HIV-1 serodiscordant couples who undergo IVF-ICSI to avoid disease transmission experience high rates of success, but also encounter complications from assisted reproductive technology similar to traditional patients. The best candidates appear to be younger women without strong risk factors for ovarian hyperstimulation syndrome. PMID- 12909500 TI - Endometrial lesions caused by catheters used for embryo transfers: a preliminary report. AB - OBJECTIVE: To visualize by microhysteroscopy any possible lesions on the endocervix and endometrium made by the catheters commonly used for embryo transfer (ET). DESIGN: Prospective descriptive study. Tertiary fertility center (IFER). PATIENT(S): Twenty-three infertile patients underwent a mock transfer before a microhysteroscopy during the postovulatory phase (days 2-5 after ovulation) of the cycle with a Tomcat catheter (n = 5), Frydman's catheter (n = 5), Frydman's set (n = 3), or Wallace's catheter (n = 10). INTERVENTION(S): Mock ETs and subsequent mycrohysteroscopies.Visualization, description, and documentation of endocervical and endometrial lesions. RESULT(S): The lesions in all 23 patients were described and documented (tunnel-like, groove-like, punch out, crater-like). The Wallace catheter appears to be less traumatic to the endometrium (but it seems that it is important to take care to not pass the internal os with the outer sheath). The Tomcat catheter and the Frydman's set caused the more significant lesions that were observed. CONCLUSION(S): In this preliminary study, for the first time endometrial lesions caused by the ET catheters were directly visualized and documented. Some of these observed lesions appear to be capable of compromising the success of ET. PMID- 12909501 TI - Differences in number and activity of peripheral natural killer cells in primary versus secondary recurrent miscarriage. AB - OBJECTIVE: To compare peripheral natural-killer (NK) cell numbers and activity in women with primary recurrent miscarriage, secondary recurrent miscarriage and controls. DESIGN: Observational study. Academic medical center. PATIENT(S): Thirty-eight women with primary recurrent miscarriage, 29 women with secondary recurrent miscarriage, and 25 control women. INTERVENTION(S): None.[1] The proportion of NK cells in the total lymphocyte population, [2] the concentration of NK cells per microliter of blood, and [3] NK activity (NKA), using both standard and whole-blood assays. RESULT(S): Primary aborters had the highest proportion and concentration of NK cells and had the highest activity using the standard assay. Secondary aborters had an intermediate level of these NK cell indices, whereas the control patients had the lowest levels. Using the whole blood NKA assay, the differences between primary and secondary aborters were most apparent: primary aborters had significantly higher NKA than did either secondary aborters or control women (72, 40, and 35 lytic units, respectively). Approximately 50% of the variability in NKA could be attributed to differences in concentrations of NK cell per microliter of blood. CONCLUSION(S): The higher NKA evident in primary recurrent miscarriage and the reported higher efficacy of immunotherapy in primary aborters support the involvement of NK cells in the etiology of primary recurrent miscarriage. PMID- 12909502 TI - A randomized, double-blind, placebo-controlled trial of heparin and aspirin for women with in vitro fertilization implantation failure and antiphospholipid or antinuclear antibodies. AB - OBJECTIVE: To investigate whether heparin and low-dose aspirin increase the pregnancy rate in antiphospholipid antibody or antinuclear antibody-seropositive women with IVF implantation failure. DESIGN: A double-blind, randomized, transfer by-transfer of fresh or cryopreserved embryos, crossover trial.A hospital infertility clinic and associated IVF service. PATIENT(S): Women seropositive for at least one antiphospholipid (APA), antinuclear (ANA), or beta(2) glycoprotein I autoantibody and >or=10 embryos transferred without achieving pregnancy (n = 143). INTERVENTION(S): Subcutaneous unfractionated heparin (5000 IU b.i.d.) and aspirin (100 mg daily) (158 transfers of 296 embryos) or placebo (142 transfers of 259 embryos) from the day of embryo transfer. MAIN OUTCOME MEASURE(S): Fetal heart per embryo transferred (implantation rate). RESULT(S): There was no significant difference in pregnancy rates or implantation rates between treated and placebo cycles; for example, fetal hearts per embryo transferred implantation rates were 6.8% (20/296) and 8.5% (22/259), respectively, and the generalized estimating equation covariate adjusted relative pregnancy rate was 0.65 (95% confidence interval, 0.33-1.28). The implantation rate for seropositive trial participants (42/555, 7.6%) compared favorably with that for IVF implantation failure patients continuing treatment outside the trial (147/3237, 4.5%). CONCLUSION(S): Heparin and aspirin did not improve pregnancy or implantation rates for APA-positive or ANA-positive patients with IVF implantation failure. PMID- 12909503 TI - Normal range and variation of the zona pellucida-induced acrosome reaction in fertile men. AB - OBJECTIVE: To investigate the normal range of the zona pellucida (ZP)-induced acrosome reaction (AR) in fertile men. DESIGN: A study of normative data. SETTING: Academic research and teaching tertiary hospital.None.Fertile men (n = 111) who had children or whose female partner was currently pregnant. MAIN OUTCOME MEASURE (S): Semen analysis, sperm-ZP binding, and the ZP-induced AR. RESULT(S): There was wide range for the ZP-induced AR: 20%-98% with a mean of 48%. The ZP-induced AR was significantly correlated with sperm concentration and normal morphology. The ZP-induced AR was significantly lower in men with sperm concentration 60 x 10(6)/mL. Similarly, men with normal sperm morphology >or=15% had significantly higher ZP-induced AR than men with normal morphology <15%. CONCLUSION(S): There is a wide range of the ZP-induced AR in fertile men that is correlated with sperm concentration and normal morphology. PMID- 12909504 TI - Comparison of controlled ovarian stimulation with human menopausal gonadotropin or recombinant follicle-stimulating hormone. AB - OBJECTIVE: To carefully examine the features of controlled ovarian stimulation performed with recombinant FSH-alpha or hMG. DESIGN: Controlled, prospective, randomized comparison of fixed gonadotropin regimens. SETTING: Academic research institution. PATIENT(S): Fifty infertile patients who were candidates for IUI. INTERVENTION(S): Patients were randomized to receive a fixed regimen of recombinant FSH-alpha (150 IU/day, 25 patients) or hMG (150 IU/day, 25 patients), after GnRH-agonist suppression (long regimen). MAIN OUTCOME MEASURES: Daily measurements of serum LH, immunoreactive FSH, hCG, E(2), P, and T. Transvaginal pelvic ultrasound every 2 days. Pregnancy and abortion rates. Cost of medications. Two recombinant FSH-alpha-treated patients did not respond. Despite matched daily FSH dose, duration of treatment (hMG 10.8 +/- 0.4 vs. recombinant FSH-alpha 12.4 +/- 0.5 days), gonadotropin dose (21.7 +/- 0.8 vs. 25.3 +/- 1.3 ampoules), gonadotropin cost (288 +/- 10 vs. 1,299 +/- 66 /cycle), serum P levels, and small preovulatory follicle number were significantly lower, and LH, hCG, immunoreactive FSH levels, and larger follicles on day 8 were significantly higher in hMG-treated patients. The pregnancy, abortion, and twin pregnancy rates did not differ. CONCLUSION: The hMG administration was associated with: [1]. increased serum LH activity and immunoreactive FSH levels during treatment; [2]. reduced signs of premature luteinization; [3]. differential modulation of folliculogenesis; [4]. lower treatment duration, gonadotropin dose, and cost; and [5]. clinical outcome comparable to recombinant FSH-alpha. PMID- 12909505 TI - Persistent osteopenia in ballet dancers with amenorrhea and delayed menarche despite hormone therapy: a longitudinal study. AB - OBJECTIVE: To investigate the role of estrogen deprivation and replacement in amenorrheic and nonamenorrheic dancers on hormone therapy and calcium. DESIGN: Clinical, placebo-controlled, randomized trial study.Healthy volunteers in an academic research environment. PATIENT(S): Fifty-five dancers (mean age: 22.0 +/- 4.6, age at menarche: 14.7 +/- 2.3 years), including 24 amenorrheics. INTERVENTION(S): Amenorrheics were randomized in a controlled trial to receive placebo or Premarin, 0.625 mg for 25 days monthly, with Provera, 10 mg, for 10 of these 25 days (hormone therapy) for 2 years. These women were compared to normally menstruating controls. The study participants also received 1250 mg of calcium per day. MAIN OUTCOME MEASURE(S): Bone mineral density (BMD) measured at the foot, wrist, and lumbar spine. Our overall results showed no difference in BMD between the treated or placebo groups, indicating that hormone therapy did not change or normalize BMD when compared to normals. Five patients (all on placebo) who resumed menses during the study showed an increase in BMD without normalization. CONCLUSION(S): These findings suggest that mechanisms other than hypoestrogenism may be involved with the osteopenia associated with exercise induced amenorrhea. PMID- 12909506 TI - Social stigma and compounded losses: quality-of-life issues for multiple-birth families. AB - OBJECTIVE: To determine the quality-of-life domains most impacted by multiple births. DESIGN: Focus groups, qualitative research. SETTING: Human volunteers in a medical research environment.Forty-three mothers, 29 raising multiple-birth children, 13 raising singletons, identified from random and convenience samples. INTERVENTION: None. MAIN OUTCOME MEASURE(S): Maternal self-reports of the psychosocial sequelae of multiple or singleton births, based on qualitative data analysis of transcribed group discussions. RESULT(S): The quality-of-life domains that were most impacted by raising multiple birth children were social stigma, pregnancy loss, marital satisfaction, children's health, unmet family needs, parenting stress, maternal depression, and the infertility experience. CONCLUSION(S): Qualitative methods identified two novel quality-of-life domains in iatrogenic multiple birth families: social stigma and compounded losses. An unexpected finding was the potential for increased marital solidification as parents coped with the inordinate stresses of multiple births. As anticipated, children's health, unmet family needs, maternal depression, and parental stress were key areas of concern. In addition, the infertility experience had a lasting impact. These findings are significant, given that at least 38% of all assisted conceptions result in a multiple birth. This study lays the groundwork for further research on the impact of iatrogenic multiple births. PMID- 12909507 TI - Peroxisome proliferator-activated receptor-gamma ligand inhibition of RANTES production by human endometriotic stromal cells is mediated through an upstream promoter element. AB - OBJECTIVE: To investigate the effect of peroxisome proliferator activated receptor-gamma (PPAR-gamma) ligands on transcription and secretion of regulated upon activation normal T-cell expressed and secreted (RANTES) in endometriotic stromal cells. DESIGN: Controlled laboratory study. SETTING: Academic research laboratory. Women in the follicular phase of the menstrual cycle undergoing laparoscopic resection for endometriosis. [1]. Transient transfection of endometriotic stromal cells with RANTES promoter vectors with and without a mutagenized PPAR-gamma response element (PPRE), then treatment with PPAR-gamma ligands; [2]. co-incubation of cells with PPAR-gamma ligands. MAIN OUTCOME MEASURE(S): RANTES promoter activity and RANTES secretion. In endometriotic stromal cells, addition of PPAR-gamma ligands (rosiglitazone and 15 deoxy Delta(12,14) prostaglandin J(2)) inhibited RANTES promoter activity by 51% and 50%, respectively. In cells transfected with the same promoter after site directed mutagenesis of the 5' PPRE, addition of PPAR-gamma ligands failed to inhibit promoter activity. When endometriotic stromal cells were treated with PPAR-gamma ligands, a decrease in RANTES secretion by 51% and 20%, respectively, was observed. CONCLUION(S): The PPAR-gamma ligands inhibit RANTES transcription and protein production in endometriotic stromal cells. Transcriptional repression appears to be mediated through a specific PPRE at -344 to -322 bp upstream from the RNA polymerase start site. PMID- 12909508 TI - Parameters affecting successful transplantation of frozen-thawed human fetal ovaries into immunodeficient mice. AB - OBJECTIVE: To compare the development and survival of human fetal follicles frozen-thawed with dimethylsulfoxide (DMSO) and propandiol (PROH) in immunodeficient mice, to study the effects of host treatment with FSH, and to compare kidney and subcutaneous transplantation. DESIGN: Controlled histologic study. SETTING: Major tertiary care and referral academic center.Twenty-one women undergoing second-trimester pregnancy termination. Microscopic morphometric analysis and immunocytochemistry for proliferating-cell nuclear antigen in human fetal ovaries grafted into immunodeficient mice. RESULTS: Renal grafts that were frozen-thawed with DMSO rather than PROH survived better in the hosts (79.6% compared with 58.8%), but significantly more follicles were identified in grafts frozen-thawed with PROH (P<.001). Follicular development was observed only in FSH treated hosts, and follicular survival and development was better in the kidney than the subcutaneous site. CONCLUSION(S): This is the first report showing development of human fetal follicles in immunodeficient mice. Freezing-thawing with PROH seems to support development and survival better than with DMSO. The kidney is a better transplantation site than the subcutaneous site, probably because of its superior vascularization. Administration of FSH to the host is essential for follicular development. Follicular development and growth was better in ovarian grafts from older fetuses, as they contained more formed follicles. PMID- 12909509 TI - Laparoscopy-assisted transinguinal extracorporeal gonadectomy in six patients with androgen insensitivity syndrome. AB - OBJECTIVE: To describe a new surgical technique to treat prophylactic gonadectomy in patients with androgen insensitivity syndrome. DESIGN: Prospective study. University hospital. PATIENT(S): Six patients with androgen insensitivity syndrome.Laparoscopy-assisted transinguinal or suprapubic extracorporeal gonadectomy. MAIN OUTCOME MEASURE(S): Accessibility, operation time, and cost effectiveness. INTERVENTION(S): Laparoscopy-assisted transinguinal extracorporeal gonadectomy under general anesthesia was performed successfully, with no complications, in all patients as a one-day procedure. The operation times were 12 to 22 minutes (average, 16 minutes). Histopathologic examination revealed testicles in all cases. The cost of each operation was approximately dollar 500 to dollar 1000. CONCLUSION(S): Laparoscopy-assisted transinguinal or suprapubic and extracorporeal gonadectomy has several advantages over laparotomy and operative laparoscopy, including short operating time, safety, simplicity, and cost-effectiveness. PMID- 12909510 TI - Factors affecting the pregnancy rate after microsurgical reversal of tubal ligation. AB - OBJECTIVE: To evaluate the factors affecting the pregnancy rate after microsurgical reversal of tubal ligation. DESIGN: Retrospective clinical study. SETTING: Private practice affiliated with a tertiary care center. One hundred twenty-eight consecutive patients who underwent tubal reversal between October 1992 and May 2001. INTERVENTION(S): Microsurgical tubal reanastomosis performed by a single surgeon. MAIN OUTCOME MEASURE(S): Subsequent pregnancy rates were evaluated with Fisher's exact tests and logistic regression according to clinical characteristics of patients. RESULT(S): The pregnancy rate was 85.7% (54 out of 63) in patients 35 years. The odds ratio (OR) between the two age groups was 7.20, with a 95% confidence interval (CI) of 2.41 to 21.55. The pregnancy rate was 85.4% (35 out of 41) in patients with body mass index (BMI) 25 (OR 3.02; CI 1.04 to 8.77). Patients sterilized 8 years (OR 3.63; CI 1.19 to 11.09). CONCLUSION(S): Age was the primary statistically significant factor affecting pregnancy rate in tubal reversal patients. Body mass index and duration of sterilization had smaller, but statistically significant, associations with pregnancy rate. PMID- 12909511 TI - Effectiveness of autocrosslinked hyaluronic acid gel after laparoscopic myomectomy in infertile patients: a prospective, randomized, controlled study. AB - OBJECTIVE: To assess the efficacy of autocrosslinked hyaluronic gel in postsurgical adhesion prevention after laparoscopic myomectomy. DESIGN: Prospective, randomized, controlled study. SETTING: University of Naples "Federico II".Thirty-six infertile women with symptomatic myomas were randomly divided into two groups of 18 patients each. INTERVENTION(S): Laparoscopic myomectomy with subserous sutures or interrupted figure 8 sutures, with (group A) or without (group B) application of autocrosslinked hyaluronic acid (HA) gel. MAIN OUTCOME MEASURE(S): Rate of postsurgical adhesions at 60-90 days of follow up. The rate of subjects who developed postoperative adhesions was significantly lower in group A in comparison with group B (27.8% vs. 77.8%). In both groups, the rate of adhesions was significantly higher in patients treated with interrupted figure 8 sutures than with subserous sutures. CONCLUSION(S): Autocrosslinked HA gel is a promising resorbable agent barrier for the reduction of postoperative adhesions after laparoscopic myomectomy. Moreover, the type of suture is a factor influencing the postsurgical adhesion formation. PMID- 12909512 TI - Percutaneous oocyte retrieval from an inguinal ovary. PMID- 12909513 TI - Pregnancies and births achieved from in vitro matured oocytes retrieved from poor responders undergoing stimulation in in vitro fertilization cycles. AB - OBJECTIVE: To describe pregnancies that resulted from in vitro matured oocytes derived from stimulated IVF cycles before cancellation owing to poor response of gonadotropins. DESIGN: Case report. University hospital. PATIENT(S): Eight patients who underwent in vitro maturation.Immature oocyte retrieval, in vitro maturation of immature oocytes, fertilization, and ET. Luteal support with progesterone and plvyeron was given. MAIN OUTCOME MEASURE(S): Pregnancy and live birth. RESULT(S): Three pregnancies (two live births and another ongoing) were achieved after immature oocyte retrieval, in vitro maturation, fertilization with ICSI, and ET. CONCLUSION(S): Immature oocyte retrieval from poor responders during stimulation, followed by in vitro maturation, may be an alternative before the cycle is canceled. PMID- 12909514 TI - Spontaneous expulsion of three large fibroids after uterine artery embolization. AB - OBJECTIVE: To report a case of uneventful expulsion of huge fibroids after uterine artery embolization. Case report. SETTING: The Department of Obstetrics and Gynaecology of a university hospital. PATIENT(S): A 45-year-old woman who underwent uterine artery embolization for fibroids. INTERVENTION(S): Transfemoral selective bilateral uterine artery catheterization and injection of 500-700 microm polyvinyl alcohol particles. RESULT(S): Recovery of the patient. The spontaneous expulsion of three fibroids on three different occasions over several months resulted in a significant reduction in menstrual loss and dysmenorrhea. CONCLUSION(S): Delivering fibroids or sections of fibroids may be a natural process after uterine artery embolization, therefore it is essential to warn women about the possible risk. Close follow-up is also essential. The size of the fibroids discharged did not require hysterectomy. Adequate antibiotic cover may be necessary to prevent sepsis. PMID- 12909515 TI - Laser-assisted human embryo biopsy on the third day of development for preimplantation genetic diagnosis: two successful case reports. AB - OBJECTIVE: To perform preimplantation genetic diagnosis (PGD) with 1.48-microm infrared diode laser assistance during embryo biopsy for two patients undergoing IVF. DESIGN: Case reports. SETTING: Private ART laboratory. Two couples undergoing IVF for infertility therapy, both of whom had previously delivered offspring afflicted with spinal muscular atrophy (type 1) after IVF therapy, and who underwent subsequent cycles of IVF coupled with PGD to screen for this disorder. INTERVENTION(S): Two individual IVF cases involving intracytoplasmic sperm injection (ICSI), embryo biopsy with laser assistance, and PGD. The ease and apparent safety of human embryo biopsy using a 1.48-microm infrared laser for partial zona pellucida (ZP) dissection to assist with embryo blastomere biopsy was evaluated. RESULT(S): Both couples were deemed to have some unafflicted embryos for transfer on the fifth day of development after blastomere biopsy in conjunction with PGD. Patient A had a singleton pregnancy and delivered a healthy normal singleton male. Patient B had a twin pregnancy; however, one twin was spontaneously lost at 10 weeks but she ultimately delivered a healthy normal singleton male. CONCLUSION(S): These successful outcomes help to demonstrate the efficacy and safety of laser-assisted embryo biopsy to facilitate PGD screening. PMID- 12909516 TI - Early effects of metformin on glucose dynamics in women with oligoamenorrhea and regular menstrual cycles who were wearing a subcutaneous glucose sensor. PMID- 12909518 TI - Culture conditions and not prolonged culture time are responsible for monozygotic twinning in human in vitro fertilization. PMID- 12909517 TI - Effects of ascorbic acid supplementation on serum progesterone levels in patients with a luteal phase defect. PMID- 12909519 TI - Abnormal endometrial development occurs during the luteal phase of nonsupplemented donor cycles treated with recombinant follicle-stimulating hormone and gonadotropin-releasing hormone antagonists. PMID- 12909520 TI - Preimplantation genetic diagnosis as both a therapeutic and diagnostic tool in assisted reproductive technology. PMID- 12909521 TI - Sex preselection and full disclosure. PMID- 12909524 TI - Links between play and dominance and attachment dimensions of dog-human relationships. AB - It is often claimed that certain behavioral problems in domestic dogs can be triggered by the games played by dog and caregiver (owner). In this study, we examine possible links between the types of games played and dimensions of the dog-owner relationship that are generally considered to affect such problems. Fifty dog-owner partnerships were filmed during 3-min play sessions in which the owner was allowed to choose the games played. All partnerships then undertook a 1 hr test designed to measure elements of behavior commonly ascribed to "dominance" and "attachment." Principal components analysis of the data produced 2 dominance related factors (Amenability and Confident Interactivity) and 4 factors describing aspects of attachment (Nonspecific Attention Seeking, Preference for Owner, Preference for Unfamiliar Person, and Separation-Related Behavior). Amenability, in particular, varied significantly between breeds. In the study, we then compared types of games played to each of these factors. Dogs playing rough and-tumble scored higher for Amenability and lower on Separation-Related Behavior than did dogs playing other types of games. Dogs playing tug-of-war and fetch scored high on Confident Interactivity. Winning or losing these games had no consistent effect on their test scores. If the dog started the majority of the games, the dog was significantly less amenable and more likely to exhibit aggression. The results suggest that how dogs play reflects general attributes of their temperament and relationship with their owner. This study provides no evidence that games play a major deterministic role on dominance dimensions of dog-human relationships, but the results suggest that playing games involving considerable body contact may affect attachment dimensions. PMID- 12909525 TI - Methodologies for the care, maintenance, and breeding of tropical poison frogs. AB - The Biodiversity Laboratories at Saint Joseph's University house more than 100 exotic and endangered animal species-including insects, fish, amphibians, and reptiles. Research in the laboratories is devoted, primarily, to understanding the genetic and ecological bases of animal behavior. In addition, a common theme of the work is the development of techniques for the successful care of exotic animals within a laboratory setting. Such techniques may be critical to studies aimed at understanding the life histories of endangered species and saving them from extinction. Current projects to study the biology of tropical poison frogs have methodologies to improve captive care, maintenance, and breeding. In this article, I present these methodologies and their impact on the welfare of captive poison frogs. PMID- 12909526 TI - The European Union ban on conventional cages for laying hens: history and prospects. AB - Since the publication of Animal Machines (Harrison, 1964), there has been widespread public pressure in Europe--supported by European institutions--to "ban the battery cage." The European Union (EU) and national governments (particularly in Northern Europe) funded research on noncage systems for egg production and enriched cages. In 1986, the EU passed a Directive specifying a minimum size for cages, but public opinion--again particularly in the North--continued to require more. A market sector emerged that would pay more for noncage eggs. Denmark, Sweden, and Switzerland passed more stringent legislation than the rest of Europe. A 1999 Directive with details based on advice from the EU's Scientific Veterinary Committee will phase out conventional laying cages but allow enriched cages. Implementation depends on various factors, including negotiations in the World Trade Organization. In the next 10 years, however, major changes to the housing of most laying hens in Europe almost certainly will occur. Similar changes in other countries will follow. As in Europe, change probably will be piecemeal, affected both by public pressure and by all sectors of society: producers, retailers, consumers, legislators, and the media. PMID- 12909527 TI - Compassion for animals in the laboratory: impairment or refinement of research methodology? AB - There are now signs in the United States as well as in Europe that the importance of a positive human-nonhuman animal relationship in research laboratories is appreciated more seriously. In addition to knowledge and skills, primary attributes of animal research personnel must be feelings of compassion and sensitivity toward animals to safeguard the reliability of scientific research data. PMID- 12909528 TI - Some reflections on the Fourth World Congress on Alternatives and Animal Use in the Life Sciences, New Orleans, Louisiana, August 11 through 15, 2002. PMID- 12909529 TI - [Abstracts of the XXIIIth Cancer Meeting. 8-10 July 2003, Paris, France]. PMID- 12909530 TI - Exposures to lead, metals, and wood dust during stripping and refinishing furniture. PMID- 12909531 TI - Exposure to trichloroethylene in an insignia manufacturing facility. PMID- 12909532 TI - Reducing risk of musculoskeletal disorders through the use of rebar-tying machines. PMID- 12909534 TI - Folpet permeation through nitrile gloves. AB - The aim of this study was to investigate whether two different brands of unsupported and unlined nitrile gloves protected against aqueous emulsions of a Folpet wettable powder (50% Folpet) using an ASTM type-I-PTC 600 permeation cell at 30.0 +/- 0.1 degrees C held in a shaking water bath. An analytical method to determine Folpet using the internal standard method was first developed based on gas chromatography-mass spectrometry (GC-MS), and gas chromatography-electron capture detection (GC-ECD). A novel pyrolysis GC-ECD technique that quantified the thermal degradation product phthalimide had pg sensitivity suitable to detect the trace amounts of Folpet that permeated. The on-column conversion was (68.0 +/ 9.5) percent at 170 degrees C over the folpet injected mass range of 3 to 148 pg. The challenge solution in the permeation cell was 1.4 mg/mL aqueous emulsion of Folpet wettable powder, and 2-propanol was the collection solvent. After evaporation of the collection solvent, the time weighted average rate of permeation of Folpet through SafeSkin nitrile (an exams type of glove) after 8 hours was (42.1 +/- 2.9) ng/cm(2)/min compared with (2.04 +/- 0.69) ng/cm(2)/min for the Sol-Vex nitrile (industrial chemical resistant), the latter being about 21 times more protective and also near the limits of detection. The respective values after 4 hours of exposure were (28.4 +/- 1.2) and (0.65 +/- 0.36) ng/cm(2)/min. Diagnostic reflectance infrared minima of both challenge and collection sides of the gloves showed small changes in wave number and intensity values after 8 hours of exposure, with Folpet being detected in dried spots on the challenge side. GC-ECD-based permeation and IR reflectance data indicated high chemical resistance of the Sol-Vex gloves to an aqueous emulsion of Folpet. PMID- 12909536 TI - Critical literature review of determinants and levels of occupational benzene exposure for United States community-based case-control studies. AB - This article presents the results of an extensive literature review identifying the uses or occurrences of, and exposures to, benzene in a variety of industries for a community-based case-control study of childhood brain cancer in the United States and Canada. We focused on industries for which quantitative exposure data were identified in studies conducted in North America in the 1980s. Each industry was coded according to the 1987 Standard Industrial Classification (SIC) system. For each industry, information relevant to exposure assessment, including process descriptions, job titles, tasks, and work practices, was summarized when available. Estimates of probability and intensity of exposure, and our confidence in these estimates are presented. Arithmetic means (AMs), weighted for the number of measurements for each industry, were calculated based on measurement data from long-term (i.e., 60+ minutes) personal sampling; short-term or area samples were only used when no other data were available for a given industry. Industries for which no quantitative exposure levels were identified in the North American literature but for which information was found on benzene use are briefly described. Published exposure data indicate that workers in most industries in the 1980s experienced exposure levels below the current standard of 1 part per million (ppm), with a weighted AM of 0.33 ppm across all industries. Despite the longtime recognition of the hematological effects of benzene, little information was available on exposure levels and determinants for many industries with potential exposure. Nevertheless, this review may clarify some of the procedures involved in assessing occupational exposures in community-based studies and may aid in the interpretation of previous occupational studies that relied on job title or industry. PMID- 12909535 TI - Impact of dermal absorption factors in occupational exposure assessment: comparison of two models for agricultural reentry workers exposed to azinphosmethyl. AB - This analysis compares two deterministic reentry exposure models that differ in their treatment of the time dependence of dermal absorption. The first model, called the "traditional model," assumes that dermal absorption is a fixed fraction of the cumulative load on skin at the end of the workshift and that absorption is independent of residence time on the skin. The second model, called the "time-integrated model," incorporates the time dependence of both exposure and absorption by assuming that absorption begins at the outset of exposure and continues through the workshift and beyond, until an effective washing event occurs. These two models were evaluated using previously collected biological monitoring data from apple thinners exposed to azinphosmethyl. Daily doses predicted by the models were compared to doses estimated from the biological sampling results assuming pseudo steady-state excretion. The geometric mean dose estimated from the biological sampling data was 20 microg/kg/day. Corresponding geometric mean doses produced by the traditional model and the time-integrated model were 79 microg/kg/day and 24 microg/kg/day, respectively. When the doses predicted by the traditional model were plotted against those estimated from the biological measurements, the slope of the regression line was significantly greater than 1 (beta = 1.37). However, when this same analysis was conducted for the doses predicted by the time-integrated model, the confidence interval around the slope encompassed 1 (beta = 1.01). Thus, time-integrated treatment of absorption appeared to provide more realistic dose estimates than did the traditional approach. PMID- 12909537 TI - Field comparison of inhalable and total dust samplers for assessing airborne dust in swine confinement barns. AB - Inhalable and total dust sampling devices were compared for evaluating airborne dust in swine confinement buildings. Measurements from three swine facilities (n = 77 paired means) were obtained by area sampling using the IOM (Institute of Occupational Medicine, Edinburgh, U.K.) inhalable dust sampler and a 37-mm closed face total (TCF) dust sampler. The overall geometric mean IOM concentration (1.18 mg/m(3), geometric standard deviation [GSD] = 2.00) was significantly greater (P < 0.05) than the overall geometric mean TCF concentration (1.08 mg/m(3), GSD = 1.98). Regression analysis with IOM and TCF values as independent and dependent variables, respectively, yielded a factor of 0.86 (+/-0.04 95% confidence interval), which can be used to estimate TCF values from the IOM measurements. Additional paired sampling data were obtained to compare the following pairs of dust samplers: (1) IOM sampler and conical inhalable sampler (CIS) (n = 20 paired means), (2) IOM and open-face total (TOF) dust samplers (n = 14), (3) CIS and TCF samplers (n = 19), and (4) TCF and TOF samplers (n = 8). Paired t-tests showed significantly (P < 0.05) higher IOM concentrations than the CIS sampler; no significant difference (P > 0.05) was found for the other three pairs compared. It may be necessary to establish work-specific conversion coefficients to obtain a reasonable estimate of worker exposure to total dust from measurements using other types of dust sampling devices. PMID- 12909538 TI - An evaluation of irritant smoke to detect exhalation valve leakage in respirators. AB - This study evaluated the ability of a qualitative fit-test method (irritant smoke) to detect known exhalation valve leakage. The OSHA protocol for the irritant smoke test mandates the use of a low flow air pump at 200 mL/minute or an aspirator squeeze bulb. Many commercial test kits include an aspirator bulb, which is subject to variation in frequency, depth of squeeze, fatigue rate, and individual hand strength. Previous studies on irritant smoke used a handheld squeeze bulb. This study evaluated the effectiveness of a low flow pump for irritant smoke fit-testing. Twenty subjects wearing North 7600 series full-face respirators equipped with P100 filters were fit-tested with a Portacount Plus to ensure adequate fit. After successful fit was demonstrated, the exhalation valve was replaced with a damaged valve and/or rotated approximately 90 degrees to produce a fit factor below 100. Having induced an exhalation valve leak, the irritant smoke fit-test was performed using the OSHA irritant smoke protocol. To avoid introducing additional unknown leakage, all head movement exercises were replaced with the head straight, normal breathing maneuver. Irritant smoke did not detect 40 percent of respirators with leaking exhalation valves. Sixty percent of the subjects were able to detect the irritant smoke. Test sensitivity was 60 percent, well below the recommended 95 percent criterion. Of the 12 subjects that detected irritant smoke, none detected the smoke in less than a minute; the average detection time was 3 min 5 s. Some subjects were able to suppress the cough reflex. These findings suggest that qualitative fit-testing using irritant smoke with a 200 ml/min continuous flow pump does not have adequate sensitivity to detect fit factors less than 100. PMID- 12909539 TI - Summary of historical beryllium uses and airborne concentration levels at Los Alamos National Laboratory. AB - Beryllium operations and accompanying medical surveillance of workers at Los Alamos National Laboratory began in the 1940s. In 1999 a Former Workers Medical Surveillance Program that includes screening for chronic beryllium disease was initiated. As part of this program, historical beryllium exposure conditions were reconstructed from archived paper and electronic industrial hygiene data sources to improve understanding of past beryllium uses and airborne concentration levels. Archived industrial hygiene sampling reports indicated beryllium was principally used in technical areas-01 and -03, primarily being machined. Beryllium was also used at 15 other technical areas in activities that ranged from explosives detonation to the manufacture of X-ray windows. A total of 4528 personal breathing zone and area air samples for beryllium, combined for purposes of calculating summary statistics, were identified during the records review phase. The geometric mean airborne beryllium concentration for the period 1949 1989 for all technical areas was 0.04 microg Be/m(3) with 97 percent of all sample below the 2.0 microg Be/m(3) occupational exposure limit (OEL). Average beryllium concentrations per decade were less than 1 microg Be/m(3) and annual geometric mean concentrations in technical area-03, the largest user of beryllium, were generally below 0.1 microg Be/m(3), indicating exposure was generally well-controlled, that is, below the OEL. Typical of many retrospective exposure assessments, not all archived data could be extracted and summarized. Despite this, we report a reasonable summary of potential beryllium uses and airborne concentration levels a worker may have encountered from 1949-1989. These data can be used to more effectively identify former worker populations at potential risk for chronic beryllium disease and to offer these workers screening as part of the Former Worker Medical Surveillance Program, and in the event that a case is diagnosed, help to understand historical exposure conditions. PMID- 12909541 TI - Ultrasonic atomization and subsequent polymer desolvation for peptide and protein microencapsulation into biodegradable polyesters. AB - Peptide and protein microencapsulation into poly(lactide) (PLA) and poly(lactide co-glycolide) (PLGA) microspheres continues to represent a technological challenge in terms of product sterility and up-scaling. The primary objective of this study was to examine the feasibility of a novel method for peptide and protein entrapment into PLA and PLGA microspheres, particularly suitable for up scaling and aseptic processing. The method involves ultrasonic atomization of an organic polymer solution combined with subsequent organic solvent extraction by a hardening agent. The study evaluated the critical atomization conditions, the required molecular cohesion parameters of polymer solvents and hardening agent for particle preparation as well as the quality of entrapment and release as a function of polymer and peptide/protein type. Suitable polymer solvents and hardening agents were restricted to defined domains of fractional cohesion parameters: f(p) = 0.2-0.35 and f(h) = 0.2-0.4 for the polymer solvents, and f(p) = 0-0.1 and f(h) = 0-0.25 for the hardening agents. Microsphere size (0.1-100 micro m) was largely controlled by the viscosity of the atomized solution. Microencapsulation of the freely water-soluble bovine serum albumin and tetrapeptide thymocartin yielded modest efficiencies of 12-35%, whereas the slightly water-soluble octapeptide vapreotide pamoate was entrapped with 63-93% efficiency. Drug release was mainly governed by the polymer type, lasting over 100 days for BSA entrapped in PLA microspheres and; 20 days for vapreotide pamoate in PLGA 50 : 50 and for thymocartin in PLA. Very importantly, the novel method was readily accommodated within a laminar air-flow cabinet. Under aseptic conditions, sterile microspheres could be prepared. In conclusion, the novel method described may have potential in industrial environments. PMID- 12909542 TI - Study on in vitro release patterns of fentanyl-loaded PLGA microspheres. AB - In order to study the development of the delivery device of long-acting local anaesthetics for post-operative analgesia and control of chronic pain of cancer patient, fentanyl loaded poly(l-lactide-co-glycolide) (PLGA, molecular weight; 5000, 8000, 20,000, and 33,000 g/mole) microspheres (FMS) were studied. FMS were prepared by an emulsion solvent-evaporation method. The influence of several preparation parameters such as initial drug loading, PLGA concentrations, emulsifier concentrations, oil phase volume and mole ratio and molecular weight has been investigated on the fentanyl release patterns. Generally, the drug showed the biphasic release patterns, with an initial diffusion followed by a lag period before the onset of the degradation phase, but there were no lag times in the device. Fentanyl was slowly released from FMS over 10 days in vitro, with a quasi-zero order property. The release rate increased with increasing drug loading as well as increasing polymer concentration with a relatively small initial burst effect. From the results, FMS may be a good formulation to deliver the anaesthesia for the treatment of chronic pain. PMID- 12909543 TI - Synthesis and release studies of shikonin-containing microcapsules prepared by the solvent evaporation method. AB - Microcapsules, containing the pharmaceutical substance shikonin, were prepared by the solvent evaporation method in order to enhance shikonin stability (reduce photo-oxidation, polymerization), decrease its hydrophobicity and control its release rate. The effect of various parameters, such as type of polymer, type and concentration of surfactant, solvent volume and mastic gum (Pistacia lentiscus resin) content/concentration as core additive, on the characteristics of the produced microcapsules and the release rate of shikonin, were experimentally investigated. Among the polymers tested for matrix, ethylcellulose (EC) of viscosity 10 cp was the most successful; EC 100 cp and mastic gum result in larger/compact particles with no pores and much slower release. Sodium dodecyl sulphate (SDS) results in microcapsules with desirable morphological and physicochemical characteristics, while polyethylene glycol (PEG) and polyvinyl alcohol (PVA) are not indicated as surfactants in shikonin microencapsulation. Decreasing the solvent volume (dichloromethane) results in increased mean particle size and, thus, in slower release rate of shikonin, while the incorporation of mastic gum in the capsule core results in better control of shikonin release. Finally, the combination of EC 10 cp as matrix, mastic gum as core additive, low dichloromethane (DCM) volume and low SDS concentration results in microcapsules with the best characteristics in terms of efficiency, loading, release and particle size distribution. PMID- 12909544 TI - Influence of alginate characteristics on the properties of multi-component microcapsules. AB - A variety of sodium alginates, differing in molar mass and structural composition, have been evaluated in the preparation of multi-component microbeads and microcapsules. Bead formation occurred by gelation with calcium chloride. Capsules were produced by reacting the pre-formed beads with the oligocation poly(methylene-co-guanidine). Despite the equiponderous (1:1) mixing with a second polyanion, sodium cellulose sulphate, the influence of the alginate properties remains evident. Specifically, the effect of the chemical composition was found to be more significant than that of the molar mass for both the mechanical and transport properties. Furthermore, for alginates of 73% alpha-l guluronic acid content less shrinking was observed compared to the 38% guluronic materials. This results in the case of the same encapsulator settings in larger microsphere diameters and thicker membranes accompanied by enhanced mechanical resistance though, also, in a higher permeability for the high-G capsules. However, subsequent coating with lower molar mass alginate allows one to adjust the permeability over a broad range, suitable for cell encapsulation and immunoprotection, without compromising the durability. PMID- 12909545 TI - Properties of drug-containing spherical pellets produced by a hot-melt extrusion and spheronization process. AB - The objectives of this study were to investigate the particle size distribution, morphology and dissolution properties of spherical pellets produced by hot-melt extrusion and spheronization and to compare the properties of hot-melt extruded pellets with beads manufactured by a traditional wet-mass extrusion and spheronization method. Spherical pellets were produced by hot-melt extrusion without the use of water or other solvents. A powder blend of theophylline, Eudragit Preparation 4135 F, microcrystalline cellulose and polyethylene glycol 8000 was hot melt-extruded and the resulting composite rod was cut into cylindrical pellets. The pellets were then spheronized in a traditional spheronizer at an elevated temperature. The same powder blend was processed using conventional wet-mass techniques. Unlike wet-mass extruded pellets, pellets prepared from hot-melt extrusion displayed both a narrow particle size distribution and controlled drug release in dissolution media less than pH 7.4. Scanning electron microscopy, X-ray diffraction and porosity measurements were employed to explain the differences in drug release rates of theophylline from pellets produced by the two processing techniques. Theophylline release from the hot-melt extruded pellets was described using the Higuchi diffusion model, and drug release rates from wet-granulated and melt-extruded pellets did not change after post-processing thermal treatment. PMID- 12909546 TI - Beneficial effects of human serum albumin on stability and functionality of alginate microcapsules fabricated in different ways. AB - A key engineering challenge in designing microcapsules made from biocompatible alginate is maintaining adequate exchange of nutrients and oxygen between the entrapped cells and the environment, while simultaneously avoiding swelling and subsequent failure of the microcapsule. Approval for the use of alginate in pharmaceutical and/or biomedical applications also strictly requires that the components of the microcapsule material must meet the safety criteria of the ASTM and FDA. Incorporation of foetal calf serum (FCS) into the microcapsules for stabilization is not in accordance with the guidelines affirmed by these organizations. FCS should be substituted by microcapsule-stabilizing additives that are medically approved. In this communication, it is shown that 10% FCS can be replaced by 1% human serum albumin (i.e. by an agent for which medical approval is granted) without compromising effects on long-term in vitro stability. Furthermore, it is demonstrated that human serum albumin (HSA) significantly enhances cell survival and, particularly, insulin secretion of encapsulated rat islets over a time period of 3 weeks when incubated in culture medium. Thus, HSA-stabilized microcapsules made from UHV(Lam) alginate are apparently a promising system for immunoisolation of cells, particularly when alginate is cross-linked by injection of BaCl(2) crystals into the alginate droplets. Slight adjustments of the alginate concentration can tailor the microcapsule permeability to the released therapeutic factor. PMID- 12909547 TI - Microencapsulation of dehydroepiandrosterone (DHEA) with poly(ortho ester) polymers by interfacial polycondensation. AB - An original encapsulation process of DHEA was developed, based on the formation of poly(ortho ester) membrane from interfacial polycondensation in an oil-in-oil emulsion. First, the formation of poly(ortho ester) (POE) in solution under anhydrous conditions between a polyol, a lactide diol and a diketene acetal (3,9 diethylidene-2,4,8,10-tetraoxaspiro-[5.5]-undecane) was studied in order to determine the structural and thermal characteristics of the POE polymer. The optimization of the formation of a fine and stable emulsion with the required size distribution was performed in relation with the type of the internal and external phases, the type and the concentration of the surfactant and the stirring rate and duration. The diffusion of monomers and DHEA was evaluated by GC-MS analysis in order to determine the mechanisms of the membrane formation. Finally the synthesis of poly(ortho ester) DHEA-loaded microcapsules was performed under anhydrous conditions required by the particular synthesis of POE. Stable poly(ortho ester) microcapsules containing DHEA were obtained with particle sizes; 1 micro m. PMID- 12909548 TI - Release characteristics of polymethacrylate nanospheres containing coumarin-6. AB - Sustained release nanospheres were prepared from the polymethacrylates Eudragit S(100) and E(100) containing a water insoluble dye by a salting-out method. Coumarin-6 was used as a model for insoluble analgesics to ascertain uptake and release properties dependent on polymer characteristics and pH. Morphology and particle size were characterized by scanning electron microscopy (SEM). Particles were smooth, spherical and uniform with diameters ranging from 0.6-0.8 micro m. Yield was 38% and 86% for E(100) and S(100), respectively, and encapsulation of coumarin-6 efficiency was 58% and 75%, respectively. Coumarin-6 was stable within the polymer matrix at temperatures from -20 degrees C to 45 degrees C for 4 months. Release was most efficient from S(100) polymers in phosphate buffer at pH 7.4 and 8.0 reaching a maximum; 5 hours prior to samples at pH 7.0 and 9.0. Release was biphasic and concentration as a function of the square root of time produced linear data suggesting a Higuchi type diffusion from a polymer matrix. Release from E(100) was 65% lower than that from S(100) and was not solely dependent upon the ionization of polymer but most likely due to a combination of factors including buffer ionization. PMID- 12909549 TI - Microparticles prepared by grinding of polymeric films. AB - Microparticles were prepared by a film grinding method, whereby thin drug containing ethylcellulose films were cryogenically ground into microparticles. The particle size and shape of the microparticles could be controlled by the thickness of the films and by the milling time. The encapsulation efficiency as well as the in vitro drug release depended on the physical state of the drug in the ethylcellulose matrix (dispersed vs dissolved). Increased drug loadings and decreased particle size and film thickness increased the drug release. Microparticles prepared from cast films were more dense and had a slower drug release compared to microparticles prepared from sprayed films or from films prepared from an aqueous colloidal ethylcellulose dispersion, Aquacoat ECD. Lamination of the drug-containing film with a drug-free polymer layer on both sides resulted in a reduced drug release. Hydrophilic plasticizers acted as pore formers and accelerated drug release, while lipophilic plasticizers reduced the drug release. The solubility of the drug in the organic polymer solution was one of the main parameters to achieve high encapsulation efficiencies and extended drug release, while dispersed drug was released much faster. The drug release from microparticles prepared by film grinding was faster than from microparticles prepared by the solvent evaporation method. The faster release was attributed to the fractured surface of the ground particles. Grinding of microparticles, which were prepared by the solvent evaporation, also resulted in a faster release. PMID- 12909554 TI - The prevalence of self-reported symptoms of respiratory disease and community belief about the severity of pollution from various sources. AB - It is postulated that health effects of air pollution may be direct and indirect through people's perception about the severity of pollution and concerns over its impact on their health. A cross sectional postal survey of some 6,559 households was conducted in the area of Ellesmere Port and Neston Borough Council. A total of 3,402 (51.9%) usable questionnaires were returned and included in the subsequent analyses. Childhood asthma was associated with central heating. Adult asthma was associated with the number of people in the house who had ever smoked and 'crowding'. General adult respiratory symptoms were associated with perception of industrial air pollution and neighbour noise in univariable but not multivariable analyses. In the multivariable model number of people in the household who had ever smoked, exposure to traffic fume pollution, crowding and living in rented accommodation. This suggests a complex relationship between actual levels of pollution (though not directly measured in this study), social deprivation, socio-behavioural factors and people's perceptions about pollution. A model of the relationship of these factors is proposed and it is argued that studies of the health impact of air pollution that concentrate only on chemical exposure will be flawed unless they are placed in the context of perception and socio-behavioural factors. PMID- 12909550 TI - Microencapsulation and oxidative stability of spray-dried fish oil emulsions. AB - Emulsions of menhaden oil and sodium caseinate (NaCas) incorporating carbohydrates of varying dextrose equivalence (DE) were spray-dried to yield encapsulated fish oil powders. The effects of carbohydrate DE (5.5-38), core/wall ratio and NaCas/carbohydrate ratio on microencapsulation efficiency (ME) and oxidative stability of spray-dried emulsions were examined. The effect of alpha tocopherol or Trolox C addition on the oxidative stability of herring oil emulsions and powders was also determined. ME of fish oil powders was greater than 90% in most cases. Peroxide value (PV) of menhaden oil powders decreased on increasing the DE of carbohydrates. PV of menhaden oil powders increased as core/wall ratio increased from 0.33-1.0. NaCas/DE 28 ratio did not affect PV values of powders. The addition of alpha-tocopherol or Trolox C decreased PV throughout the storage period; this effect was most pronounced for alpha tocopherol added to the oil at a concentration of 100 ppm. Addition of alpha tocopherol delays the onset of oxidation in stored fish oil powders. PMID- 12909555 TI - Mercury in hair and in fish consumed by Riparian women of the Rio Negro, Amazon, Brazil. AB - This work puts the risks and benefits of a mercury acquisition through a fish dominated diet in the context of the other endemic health problems in the Amazon. Fish consumption rate was assessed after determining mercury concentrations in the most consumed fish and in hair of women at fertile age (15 to 45 years of age), living in remote localities of the Rio Negro basin (not impacted by gold mining). Fish-meals were consumed once a day (7.1%) but in most cases (78.6 %) at least twice a day with an estimated mean fish consumption rate of 170.5 g (range 23 to 293 g). The fish-species most preferred were Tucunares (Cichla spp), Pacus (Mylossoma aureum and Myleus micans), Piranhas (Sarrasalmus spp), Cara (Satanoperca jurupari), Jaraqui (Semaprochilodus taeniurus). The mean Hg concentration in these species varies widely (38 to 592 ng g(- 1)) but 27% of samples had Hg concentrations above 500 ng g(- 1), and only 7% were above 1000 ng g(- 1). The hair-Hg concentrations ranged from 6.5 to 32.6 microg g(- 1) with 82% of them having concentrations above 10 microg g(- 1) (w.w.) hair. Hair-Hg concentrations were positively correlated with women's age (r=0.4500; P=0.0111). Clinical evaluation did not detected symptoms (paraparesis, tremor, numbness of limbs, sensory disturbances) compatible with Minamata disease, but there was a high (71%) incidence of clinical history of malaria. Fish is an abundant natural resource, rich in essential nutrients and important to the diet of 'ribeirinhos' of Brazilian Amazonia. We hypothesize that lack of basic medical care in these communities outweighs risk of naturally occurring fish-MMHg. Therefore food habits based on fish consumption is part of a successful strategy for survival and should be considered as a health asset. PMID- 12909556 TI - Validation of a questionnaire and microenvironmental model for estimating past exposures to ozone. AB - Epidemiology studies of the health effects associated with chronic exposure to ozone are constrained by a lack of validated exposure assessment methods that account for activity patterns and building ventilation, two potentially important predictors of ozone exposure. A pilot study was conducted to evaluate the impact of these parameters on an exposure model, to evaluate how reliably they can be reconstructed, and how this reliability affects estimates of past exposures to ozone. Fourteen subjects completed time-activity diaries and wore personal passive monitors during summer 1996 and reconstructed their activities 1 year later in a questionnaire. These data were used in a microenvironmental exposure model and the results were compared to measured personal exposure. A model using activity patterns, building ventilation, and location-specific ambient ozone data produced mean exposure estimates 18.9 +/- 1.5 ppb ozone greater than personal measurements. A simpler model that omitted variation in indoor ventilation produced exposure estimates 5.0 +/- 1.3 ppb ozone greater than mean measured exposures. The questionnaire was a reliable survey method for reconstructing past activity patterns. However, detailed histories of building ventilation may not be required to reconstruct reliably past exposures to ozone. The simple model was more precise and more practical for implementation in large epidemiological studies of health effects related to chronic exposure to ambient ozone. PMID- 12909557 TI - Myocarditis: an expected health hazard associated with water resources contaminated with Coxsackie viruses type B. AB - Enteroviruses, especially Coxsackie B viruses (CBVs), are responsible for approximately 50% of cases of viral myocarditis. In the present study, serum samples (160) were collected from acute myocarditis patients at different age groups and 104 samples of the same age groups as a control. Cholesterol, LDH, CPK, and GOT were measured for all serum samples (264). Also, to study the source of virus transmission, 72 water and 72 wastewater samples were collected from water and wastewater treatment plants at intakes and outlets. Water and wastewater samples were concentrated by filtration through Zeta-plus filter cartridges and reconcentrated by the PEG-6000 precipitation method. Serum, water, and wastewater samples were inoculated in BGM cells for three successive passages. RT-PCR with enterovirus primers was carried out directly for serum samples and for 1st and 3rd cell culture passages. The positive samples were used for neutralization assay using anti-CBV sera pool to determine the CBV followed by neutralization with separate antisera. The results showed that 50 (31.25%) serum samples from acute myocarditis patients and two (1.4%) samples from the controls were positive for enterovirus RT-PCR. For water and wastewater samples enteroviruses were present in 63.8% and 8.3% for intake and outlet of water treatment plants and, 66.6% and 47.2% for intake and outlet of wastewater treatment plants, respectively. The level of CBV serotypes was varied where CBV3 was dominant for all age groups of myocarditis patients and CBV2 and CBV5 were also detected while CBV2 was the main CBV in water samples and CBV2, 3 and 5 were detected in wastewater samples. The integration of cell culture-PCR reduces the time required for virus detection and enhances the sensitivity of the test. PMID- 12909558 TI - Risk factors for increased urinary inorganic arsenic concentrations from low arsenic concentrations in drinking water. AB - A large number of drinking water supplies worldwide have greater than 50 microg l(- 1) inorganic arsenic in drinking water, and there is increasing pressure to reduce concentrations. Few studies have specifically considered low concentrations of arsenic in water supplies and the significance of other factors which may contribute to increased exposure. This study aimed to investigate risk factors for increased urinary inorganic arsenic concentrations, in a population exposed to 10 - 100 microg l(- 1) of arsenic in drinking water, as well as a control population with lower arsenic concentrations in their drinking water. Inorganic arsenic in urine was used as the measure of exposure. The median drinking water arsenic concentration in the exposed population was 43.8 microg l( 1) (16.0 - 73 microg l(- 1)) and less than the analytical limit of detection of 1 microg l(- 1) (95% (P < 0.01) of cell binding to hyaluronic acid (HA). The cytokines TNF-alpha, IFN-gamma, IL-1 beta, and IL-4 stimulated a 2- to 3.5-fold increase in CD44 dependent cell binding to HA. IFN-gamma treatment did not increase CD44 expression as assessed by flow cytometry, although phorbol myristate acetate treatment did. This indicates that IFN-gamma-induced cell binding to HA did not require increased CD44 expression. These data indicate that CD44 is important for bronchial epithelial cell binding to HA and that cytokines known to be expressed in inflammation can increase HA binding independently of the level of CD44 expression. PMID- 12909590 TI - Early high expression of IP-10 in F344 rats resistant to Sendai virus-induced airway injury. AB - Weanling F344 and BN rats differ markedly in their susceptibility to Sendai virus induced airway injury. Early gene expression that controls their differences in susceptibility remains poorly understood. In this study we combined suppressive subtractive hybridization and cDNA library array hybridization to identify genes differentially expressed in virus-susceptible BN and virus-resistant F344 rats during the first 3 days after inoculation. Differential expression of selected clones was further verified by quantitative RT-PCR. Seven virus-induced gene segments were identified. Of them, interferon-gamma-inducible protein 10 (IP-10), Mx1, and guanylate-binding protein-2 mRNA abundance in infected F344 rats was 201.5, 188.2, and 281.7% higher, respectively, than that of infected BN rats at 2 days after inoculation. In situ hybridization indicated that virus-induced IP-10 was expressed mainly in airway epithelial cells of F344 rats. Sendai virus infection can directly induce IP-10 expression in rat tracheal epithelial cells in vitro. IP-10 early high expression might contribute to the resistance to virus induced airway disease in F344 rats by promoting Th1 responses and increasing antiviral activity. PMID- 12909591 TI - Differential roles for NF-kappa B in endotoxin and oxygen induction of interleukin-8 in the macrophage. AB - The alveolar macrophage is an important source of interleukin (IL)-8 during pulmonary injury. The IL-8 gene promoter sequence contains nuclear factor (NF) kappa B, NF-IL6, and activator protein (AP)-1 binding sequences. These sites may have differing regulatory roles in hyperoxia-exposed macrophages than in those stimulated by bacterial lipopolysaccharide (LPS). U-937 and THP-1 macrophage-like cells were exposed to air-5% CO2 or 95% O2-5% CO2, with or without 1.0 microg/ml of LPS, and transfected with an IL-8 promoter-reporter containing NF-kappa B, NF IL6, or AP-1 mutations. Hyperoxia and LPS caused additive increases in IL-8 production by U-937 cells, whereas THP-1 cells responded only to LPS. An NF-kappa B mutation ablated baseline and O2- and LPS-stimulated reporter activity in both cell lines, whereas NF-IL6 mutations had little effect. An AP-1 mutation had an intermediate effect. LPS, but not hyperoxia, stimulated nuclear translocation of NF-kappa B in both cell lines. Pharmacological blockade of NF-kappa B nuclear translocation ablated LPS-, but not hyperoxia-, stimulated IL-8 production. Although an intact promoter NF-kappa B site is crucial to macrophage IL-8 production, only LPS-stimulated production appears to require additional nuclear translocation of NF-kappa B. PMID- 12909592 TI - Inhibition of alveolarization and altered pulmonary mechanics in mice expressing GATA-6. AB - GATA-6, a member of a family of zinc finger transcription factors, is expressed in epithelial cells of the developing lung. To further assess the role of GATA-6 in lung morphogenesis, GATA-6 was expressed in respiratory epithelial cells of the developing mouse lung under control of the surfactant protein C promoter (hSP CGATA-6 mice). Although GATA-6 did not alter lung morphology at embryonic day 18.5, defects in alveolar septation were observed early in the neonatal period, and air space enlargement persisted to adulthood. Airway resistance, airway elastance, tissue damping, and tissue elastance were significantly decreased, and lung volumes were significantly increased at 12 wk of age. Normal postnatal morphogenesis of the lung depends upon precise temporal-spatial regulation of GATA-6. PMID- 12909593 TI - Hypoxia increases AP-1 binding activity by enhancing capacitative Ca2+ entry in human pulmonary artery endothelial cells. AB - Activating protein (AP)-1 transcription factors modulate expression of genes involved in cell proliferation and migration. Chronic hypoxia increases pulmonary artery smooth muscle cell proliferation by upregulating AP-1-responsive genes encoding for endothelium-derived vasoactive and mitogenic factors implicated in pulmonary hypertension development. The expression of AP-1 transcription factors is sensitive to changes in cytosolic free [Ca2+] ([Ca2+]cyt). Capacitative Ca2+ entry (CCE) via store-operated Ca2+ channels (SOC) is an important mechanism for raising [Ca2+]cyt in pulmonary artery endothelial cells (PAEC). Using combined molecular biological, fluorescence microscopy, and biophysical approaches, we examined the effect of chronic hypoxia (3% O2, 72 h) on AP-1 DNA binding activity, CCE, and transient receptor potential (TRP) gene expression in human (h) PAEC. EMSA showed that AP-1 binding to hPAEC nuclear protein extracts was significantly enhanced by hypoxia, the increase being dependent on store-operated Ca2+ influx and sensitive to La3+, an SOC inhibitor. Hypoxia also increased basal [Ca2+]cyt, the amount of CCE produced by store depletion with cyclopiazonic acid, and the amplitude of SOC-mediated currents (ISOC). The increases of CCE amplitude and ISOC current density by hypoxia were paralleled by enhanced TRPC4 mRNA and protein expression. Hypoxia-enhanced CCE and TRPC4 expression were also attenuated by La3+. These data suggest that hypoxia increases AP-1 binding activity by enhancing Ca2+ influx via La3+-sensitive TRP-encoded SOC channels in hPAEC. The Ca2+-mediated increase in AP-1 binding may play an important role in upregulating AP-1-responsive gene expression, in stimulating pulmonary vascular cell proliferation and, ultimately, in pulmonary vascular remodeling in patients with hypoxia-mediated pulmonary hypertension. PMID- 12909594 TI - Neuronal sensitivity to hyperoxia, hypercapnia, and inert gases at hyperbaric pressures. AB - As ambient pressure increases, hydrostatic compression of the central nervous system, combined with increasing levels of inspired Po2, Pco2, and N2 partial pressure, has deleterious effects on neuronal function, resulting in O2 toxicity, CO2 toxicity, N2 narcosis, and high-pressure nervous syndrome. The cellular mechanisms responsible for each disorder have been difficult to study by using classic in vitro electrophysiological methods, due to the physical barrier imposed by the sealed pressure chamber and mechanical disturbances during tissue compression. Improved chamber designs and methods have made such experiments feasible in mammalian neurons, especially at ambient pressures <5 atmospheres absolute (ATA). Here we summarize these methods, the physiologically relevant test pressures, potential research applications, and results of previous research, focusing on the significance of electrophysiological studies at <5 ATA. Intracellular recordings and tissue Po2 measurements in slices of rat brain demonstrate how to differentiate the neuronal effects of increased gas pressures from pressure per se. Examples also highlight the use of hyperoxia (100-fold selective for CCR1 as compared with a panel of G-protein-coupled receptors including related chemokine receptors. Evidence for its potential use in human disease is suggested by its ability to inhibit 90% of the monocyte chemotactic activity present in 11/15 rheumatoid arthritis synovial fluid samples. These data illustrate that CP-481,715 is a potent and selective antagonist for CCR1 with therapeutic potential for rheumatoid arthritis and other inflammatory diseases. PMID- 12909631 TI - Lipid phosphate phosphatases regulate lysophosphatidic acid production and signaling in platelets: studies using chemical inhibitors of lipid phosphate phosphatase activity. AB - Blood platelets play an essential role in ischemic heart disease and stroke contributing to acute thrombotic events by release of potent inflammatory agents within the vasculature. Lysophosphatidic acid (LPA) is a bioactive lipid mediator produced by platelets and found in the blood and atherosclerotic plaques. LPA receptors on platelets, leukocytes, endothelial cells, and smooth muscle cells regulate growth, differentiation, survival, motility, and contractile activity. Definition of the opposing pathways of synthesis and degradation that control extracellular LPA levels is critical to understanding how LPA bioactivity is regulated. We show that intact platelets and platelet membranes actively dephosphorylate LPA and identify the major enzyme responsible as lipid phosphate phosphatase 1 (LPP1). Localization of LPP1 to the platelet surface is increased by exposure to LPA. A novel receptor-inactive sn-3-substituted difluoromethylenephosphonate analog of phosphatidic acid that is a potent competitive inhibitor of LPP1 activity potentiates platelet aggregation and shape change responses to LPA and amplifies LPA production by agonist-stimulated platelets. Our results identify LPP1 as a pivotal regulator of LPA signaling in the cardiovascular system. These findings are consistent with genetic and cell biological evidence implicating LPPs as negative regulators of lysophospholipid signaling and suggest that the mechanisms involve both attenuation of lysophospholipid actions at cell surface receptors and opposition of lysophospholipid production. PMID- 12909632 TI - Characterization of the intracellular transport of GluR1 and GluR2 alpha-amino-3 hydroxy-5-methyl-4-isoxazole propionic acid receptor subunits in hippocampal neurons. AB - Little is known about the dynamics of the dendritic transport of alpha-amino-3 hydroxy-5-methyl-4-isoxazole propionic acid receptors (AMPARs) to synapses. Here, using virally expressed green fluorescent protein (GFP)-GluR1 and GFP-GluR2 and confocal photobleach techniques we show near real-time movement of these subunits in living cultured hippocampal neurons. GFP-GluR1 fluorescence was widely distributed throughout the extranuclear compartment with no evidence for discrete intracellular stores. GFP-GluR1 transport was predominantly proximal to distal at rates of 0.2-0.4 mum.s-1. GFP-GluR2 fluorescence was more punctate and localized at or close to the plasma membrane. Overall, GFP-GluR2 movement was less dynamic with distinct mobile and immobile pools. Neither activation nor inhibition of surface-expressed N-methyl-d-aspartate receptors or AMPARs had any significant effect on the rates of GFP-GluR1 or GFP-GluR2 dendritic transport. These results demonstrate that GluR1 is constitutively and rapidly transported throughout the neuron. GluR2, on the other hand, is less mobile, with a majority retained in relatively immobile membrane-associated clusters, with approximately 40% showing synaptic co-localization. Furthermore, the transport of both subunits is activity independent, suggesting that the regulated delivery of AMPARs to the vicinity of synapses is not a mechanism that is involved in processes such as synaptic plasticity. PMID- 12909634 TI - YodA from Escherichia coli is a metal-binding, lipocalin-like protein. AB - We have determined the crystal structure of YodA, an Escherichia coli protein of unknown function. YodA had been identified under conditions of cadmium stress, and we confirm that it binds metals such as cadmium and zinc. We have also found nickel bound in one of the crystal forms. YodA is composed of two domains: a main lipocalin/calycin-like domain and a helical domain. The principal metal-binding site lies on one side of the calycin domain, thus making YodA the first metal binding lipocalin known. Our experiments suggest that YodA expression may be part of a more general stress response. From sequence analogy with the C-terminal domain of a metal-binding receptor of a member of bacterial ATP-binding cassette transporters, we propose a three-dimensional model for this receptor and suggest that YodA may have a receptor-type partner in E. coli. PMID- 12909633 TI - Cloning and developmental analysis of murid spermatid-specific thioredoxin-2 (SPTRX-2), a novel sperm fibrous sheath protein and autoantigen. AB - Thioredoxins compose a growing family of proteins that participate in different cellular processes via redox-mediated reactions. We report here the cloning, developmental expression, and location of murid Sptrx-2. Mouse and rat SPTRX-2 proteins display a high homology to their human ortholog in the thioredoxin and NDP kinase domains, and the coding genes are located at syntenic positions. Northern blotting and in situ hybridization confirmed the testis-specific expression of murine Sptrx-2 mRNA, mostly in round spermatids. Immunohistochemical analysis of the 19 steps of rat spermiogenesis showed that SPTRX-2 expression becomes prominent in the cytoplasmic lobe of step 15-18 spermatids and diminishes in step 19 just before spermiation. However, in the spermatid tail, SPTRX-2 immunoreactivity increased from step 15 to 19 and was confined to the principal piece. By immunogold electron microscopy, SPTRX-2 was first detected scattered throughout the cytoplasm of the axoneme in step 14-15 spermatids, but began to be incorporated by step 16 into the fibrous sheath (FS). During steps 17-18, the labeling increased over the ribs and columns of the assembled FS. It peaked in step 19 and remained in the FS of epididymal spermatozoa. Immunoblots of isolated FS obtained from spermatozoa confirmed that SPTRX-2 is an integral component of the FS and a post-obstruction autoantigen in vasectomized rats. Our data indicate that SPTRX-2 incorporation into the FS lags well behind FS assembly, suggesting it is required during the final stages of sperm tail maturation in the testis and/or epididymis, where extensive disulfide bonding of FS proteins occurs. PMID- 12909635 TI - Effects of nonlipolytic ligand function of endothelial lipase on high density lipoprotein metabolism in vivo. AB - Endothelial lipase (EL) influences high density lipoprotein (HDL) metabolism in vivo and mediates bridging and uptake of HDL particles independent of its lipolytic activity in vitro. To determine whether EL has a nonlipolytic ligand function in HDL metabolism in vivo, 1 x 1011 particles of a recombinant adenovirus encoding human EL (AdEL), catalytically inactive human EL (AdELS149A), or control (Adnull) were injected into wild-type, apoA-I transgenic, and hepatic lipase knockout mice. ELS149A protein was expressed at higher levels than wild type EL. EL and ELS149A protein were both substantially increased in the postheparin plasma compared with preheparin, indicating that both the wild-type and mutant EL were bound to cell-surface heparan sulfate proteoglycans. Overexpression of wild-type EL was associated with a significantly increased postheparin-plasma phospholipase activity and dramatically decreased levels of total cholesterol, HDL cholesterol, phospholipids, and apoA-I. Injection of AdELS149A did not result in increased phospholipase activity confirming that ELS149A was catalytically inactive. Expression of ELS149A did not decrease lipid or apoA-I levels in wild-type and apoA-I transgenic mice yet led to an intermediate reduction of total cholesterol, HDL cholesterol, and phospholipids in hepatic lipase-deficient mice compared with control and EL-expressing mice. Our study demonstrates for the first time that EL has both a lipolytic and nonlipolytic function in HDL metabolism in vivo. Lipolytic activity of EL, however, seems to be most important for its effects on systemic HDL metabolism. PMID- 12909636 TI - An essential role for aspartate 264 in catalysis by tRNA-guanine transglycosylase from Escherichia coli. AB - tRNA-guanine transglycosylase (TGT) catalyzes a post-transcriptional base exchange reaction involved in the incorporation of the modified base queuine (Q) into the wobble position of certain tRNAs. Catalysis by TGT occurs through a double-displacement mechanism that involves the formation of a covalent enzyme RNA intermediate (Kittendorf, J. D., Barcomb, L. M., Nonekowski, S. T., and Garcia, G. A. (2001) Biochemistry 40, 14123-14133). The TGT chemical mechanism requires the protonation of the displaced guanine and the deprotonation of the incoming heterocyclic base. Based on its position in the active site, it is likely that aspartate 264 is involved in these proton transfer events. To investigate this possibility, site-directed mutagenesis was employed to convert aspartate 264 to alanine, asparagine, glutamate, glutamine, lysine, and histidine. Biochemical characterization of these TGT mutants revealed that only the conservative glutamate mutant retained catalytic activity, with Km values for both tRNA and guanine 3-fold greater than those for wild-type, whereas the kcat was depressed by an order of magnitude. Furthermore, of these six TGT mutants, only the TGT(D264E) was capable of forming a TGT.RNA covalent intermediate; however, unlike wild-type TGT, only hydroxylamine is capable of cleaving the TGT(D264E).RNA covalent complex. In an effort to better understand the unique biochemical properties of the D264E TGT mutant, we solved the crystal structure of the Zymomonas mobilis TGT with the analogous mutation (D280E). The results of these studies support two roles for aspartate 264 in catalysis by TGT, protonation of the leaving guanine and deprotonation of the incoming preQ1. PMID- 12909637 TI - Glycation induces formation of amyloid cross-beta structure in albumin. AB - Amyloid fibrils are components of proteinaceous plaques that are associated with conformational diseases such as Alzheimer's disease, transmissible spongiform encephalopathies, and familial amyloidosis. Amyloid polypeptides share a specific quarternary structure element known as cross-beta structure. Commonly, fibrillar aggregates are modified by advanced glycation end products (AGE). In addition, AGE formation itself induces protein aggregation. Both amyloid proteins and protein-AGE adducts bind multiligand receptors, such as receptor for AGE, CD36, and scavenger receptors A and B type I, and the serine protease tissue-type plasminogen activator (tPA). Based on these observations, we hypothesized that glycation induces refolding of globular proteins, accompanied by formation of cross-beta structure. Using transmission electron microscopy, we demonstrate here that glycated albumin condensates into fibrous or amorphous aggregates. These aggregates bind to amyloid-specific dyes Congo red and thioflavin T and to tPA. In contrast to globular albumin, glycated albumin contains amino acid residues in beta-sheet conformation, as measured with circular dichroism spectropolarimetry. Moreover, it displays cross-beta structure, as determined with x-ray fiber diffraction. We conclude that glycation induces refolding of initially globular albumin into amyloid fibrils comprising cross-beta structure. This would explain how glycated ligands and amyloid ligands can bind to the same multiligand "cross beta structure" receptors and to tPA. PMID- 12909638 TI - Hepatitis C virus NS5A and subgenomic replicon activate NF-kappaB via tyrosine phosphorylation of IkappaBalpha and its degradation by calpain protease. AB - Hepatitis C virus nonstructural protein 5A (NS5A) has been implicated in the HCV antiviral resistance, replication, and transactivation of cellular gene expression. We have recently shown that HCV NS5A activates NF-kappaB via oxidative stress (22). In this study, we investigate the molecular mechanism(s) of NF-kappaB activation in response to oxidative stress induced by NS5A protein. In contrast to the classic Ser32,36 phosphorylation of IkappaBalpha, we report here that tyrosine phosphorylation of IkappaBalpha at Tyr42 and Tyr305 residues is induced by the HCV NS5A and the subgenomic replicons in the NF-kappaB activation process. Use of IkappaBalpha-Tyr42,305 double mutant provided the evidence for their key role in the activation of NF-kappaB. Activation of NF kappaB was blocked by a series of tyrosine kinase inhibitors but not by IkappaB kinase inhibitor BAY 11-7085. More specifically, a ZAP-70 knock-out cell line expressing NS5A and other nonstructural proteins respectively prevented the NF kappaB activation, indicating the involvement of ZAP-70 as a probable tyrosine kinase in the activation process. Evidence is also presented for the possible role of calpain proteases in the NS5A-induced IkappaBalpha degradation. These studies collectively define an alternate pathway of NF-kappaB activation by NS5A alone or in the context of the HCV subgenomic replicon. Constitutive activation of NF-kappaB by HCV has implications in the chronic liver disease including hepatocellular carcinoma associated with HCV infection. PMID- 12909639 TI - RecQ helicase stimulates both DNA catenation and changes in DNA topology by topoisomerase III. AB - Together, RecQ helicase and topoisomerase III (Topo III) of Escherichia coli comprise a potent DNA strand passage activity that can catenate covalently closed DNA (Harmon, F. G., DiGate, R. J., and Kowalczykowski, S. C. (1999) Mol. Cell 3, 611-620). Here we directly assessed the structure of the catenated DNA species formed by RecQ helicase and Topo III using atomic force microscopy. The images show complex catenated DNA species involving crossovers between multiple double stranded DNA molecules that are consistent with full catenanes. E. coli single stranded DNA-binding protein significantly stimulated both the topoisomerase activity of Topo III alone and the DNA strand passage activity of RecQ helicase and Topo III. Titration data suggest that an intermediate of the RecQ helicase unwinding process, perhaps a RecQ helicase-DNA fork, is the target for Topo III action. Catenated DNA is the predominant product under conditions of molecular crowding; however, we also discovered that RecQ helicase and single-stranded DNA binding protein greatly stimulated the intramolecular strand passage ("supercoiling") activity of Topo III, as revealed by changes in the linking number of uncatenated DNA. Together our results demonstrate that RecQ helicase and Topo III function together to comprise a potent and concerted single-strand DNA passage activity that can mediate both catenation-decatenation processes and changes in DNA topology. PMID- 12909640 TI - Protein region important for regulation of lipid metabolism in angiopoietin-like 3 (ANGPTL3): ANGPTL3 is cleaved and activated in vivo. AB - Angiopoietin-like 3 (ANGPTL3) is a secreted protein that is mainly expressed in the liver and regulates lipid metabolism by inhibiting the lipolysis of triglyceriderich lipoproteins. Using deletion mutants of human ANGPTL3, we demonstrated that the N-terminal coiled-coil domain-containing fragment-(17-207) and not the C-terminal fibrinogen-like domain-containing fragment-(207-460) increased the plasma triglyceride levels in mice. We also found that the N terminal region 17-165 was required to increase plasma triglyceride levels in mice and that a substitution of basic amino acid residues in the region 61-66 of the fragment showed no increase in the plasma triglyceride levels and no inhibition of lipolysis by lipoprotein lipase. In addition, when we analyzed ANGPTL3 in human plasma, we detected cleaved fragments of ANGPTL3. By analyzing recombinant ANGPTL3 in mouse plasma, we found that it was cleaved at two sites, Arg221 downward arrow Ala222 and Arg224 downward arrow Thr225, which are located in the linker region between the coiled-coil domain and the fibrinogen-like domain. Furthermore, a cleavage-resistant mutant of ANGPTL3 was determined to be less active than wild-type ANGPTL3 in increasing mouse plasma triglyceride levels but not in inhibiting lipoprotein lipase activity. These findings suggest that the cleavage of ANGPTL3 is important for the activation of ANGPTL3 in vivo. PMID- 12909641 TI - Steroidogenic acute regulatory protein-binding protein cloned by a yeast two hybrid system. AB - Steroidogenic acute regulatory (StAR) protein plays a key role in the transport of cholesterol from the outer mitochondrial membrane to the inner membrane. A StAR mutant protein lacking the first 62 amino acids (N-62 StAR protein) has been reported to be as effective as wild-type StAR protein. In the present study, we examined the mechanism by which StAR protein stimulates steroidogenesis. A Gal4 based yeast two-hybrid system was used to identify proteins interacting with N-62 StAR protein. Nine positive clones were obtained from screening 1 x 106 clones. The results of pull-down assays and mammalian two-hybrid assays confirmed interaction between N-62 StAR protein and the clone 4 translated product. The clone 4 translated product was named StAR-binding protein (SBP). We prepared an expression plasmid (pSBP) by inserting SBP cDNA into the pTarget vector. After cotransfection with the human cytochrome P450scc system, StAR expression vector, and pSBP, the amount of pregnenolone produced by COS-1 cells was increased. The amount of steroid hormones produced by steroidogenic cells subjected to small interfering RNA treatment was less than that produced by control cells. In conclusion, SBP binds StAR protein in cells and enhances the ability of StAR protein to promote syntheses of steroid hormones. PMID- 12909642 TI - The region between transmembrane domains 1 and 2 of the reduced folate carrier forms part of the substrate-binding pocket. AB - A functional cysteine-less form of the hamster reduced folate carrier protein was generated by alanine replacement of the 14 cysteine residues. The predicted 12 transmembrane topology was examined by replacing selected amino acids, predicted to be exposed to the extracellular or cytosolic environments, with cysteines. The location of these cysteines was defined by their accessibility to biotin maleimide in the presence or absence of specific blocking agents. Amino acids predicted to be exposed to the extracellular environment (S46C, S179C, L300C, Y355C, and K430C) could be labeled with biotin maleimide; this modification could be blocked by prior treatment with nonpermeable reagents. Amino acids predicted to be within the cytosol (S152C, Cys224, and L475C) could be labeled only after streptolysin O permeabilization. In addition, the cysteine-less reduced folate carrier was exploited to evaluate a potential substrate-binding domain as suggested by previous studies. Nineteen cysteine replacements were generated between residues 39 and 75, a region located between the first and second transmembrane segments. From the biotinylation of these sites and the ability of various reagents to block this labeling, it appears that L41C, E45C, S46C, T49C, I66C, and L70C are exposed to the extracellular environment, whereas Q54C, Q61C, and T63C are slightly less accessible. Cysteines 39, 42, 44, 47, 51, and 73 were inefficiently biotinylated, suggesting that these sites are located in the membrane or within a tightly folded domain of the protein. Furthermore, biotinylation of cysteines 41, 46, 49, 70, and 71 could be prevented by prior treatment with either methotrexate or folinic acid, indicating that these sites form part of a substrate-binding pocket. PMID- 12909643 TI - The upstream open reading frame mediates constitutive effects on translation of cytochrome p-450c27 from the seventh in-frame AUG codon in rat liver. AB - The 2.3-kb mRNA that codes for cytochrome P-450c27 (CYP27) has an unexpectedly long 5'-untranslated region (UTR) that holds six AUGs, leading to several upstream open reading frames (uORFs). The initiation of translation from the seventh AUG forms a putative 55-kDa precursor, which is processed in mitochondria to form a 52-kDa mature protein. The first three AUGs form fully overlapping uORF1, uORF2, and uORF3 that are in-frame with the seventh AUG and next two form fully overlapping uORF4 and uORF5 that are out-of-frame with the seventh AUG. Although not recognized by the scanning ribosomes under normal conditions, the sixth in-frame AUG forms a putative 57-kDa extension of the main open reading frame. The purpose of this study was to identify the elements in the 5'-UTR that direct CYP27 mRNA translation exclusively from the seventh AUG. Expression of 5' deletion mutants in COS cells reveal that the intact 5'-UTR not only directs the initiation of translation from the seventh AUG but also acts as a negative regulator. A 2-kb deletion mutant that lacks uORF1 initiates translation equally from the sixth and the seventh AUGs, forming both 57- and 55-kDa precursor proteins with a 2-fold increase in rate of translation. However, induction in translation does not affect the levels of the mature 52-kDa form in mitochondria but causes accumulation of the precursor form in cytosol not seen in COS cells transfected with wild-type cDNA. Mutation of the stop codon that terminates uORF1 completely shifts the initiation of translation from the seventh to the first AUG, forming a 67-kDa precursor that is processed into a 52-kDa mature protein in mitochondria. Confirmation of the bicistronic nature of CYP27 mRNA by epitope mapping of uORF1 suggests that translation of CYP27 mRNA from the seventh AUG is directed and regulated by uORF1 expression. PMID- 12909644 TI - Recognition of the N-terminal modules of thrombospondin-1 and thrombospondin-2 by alpha6beta1 integrin. AB - In addition to its recognition by alpha3beta1 and alpha4beta1 integrins, the N terminal pentraxin module of thrombospondin-1 is a ligand for alpha6beta1 integrin. alpha6beta1 integrin mediates adhesion of human microvascular endothelial and HT-1080 fibrosarcoma cells to immobilized thrombospondin-1 and recombinant N-terminal regions of thrombospondin-1 and thrombospondin-2. alpha6beta1 also mediates chemotaxis of microvascular cells to thrombospondin-1 and thrombospondin-2. Using synthetic peptides, LALERKDHSG was identified as an alpha6beta1-binding sequence in thrombospondin-1. This peptide inhibited alpha6beta1-dependent cell adhesion to thrombospondin-1, thrombospondin-2, and the E8 fragment of murine laminin-1. The Glu residue in this peptide was required for activity, and the corresponding residue (Glu90) in the N-terminal module of thrombospondin-1 was required for its recognition by alpha6beta1, but not by alpha4beta1. alpha6beta1 was also expressed in human umbilical vein endothelial cells; but in these cells, only certain agonists could activate the integrin to recognize thrombospondins. Selective activation of alpha6beta1 integrin in microvascular endothelial cells by the anti-beta1 antibody TS2/16 therefore accounts for their adhesion responses to thrombospondins and explains the distinct functions of alpha4beta1 and alpha6beta1 integrins as thrombospondin receptors in microvascular and large vessel endothelial cells. PMID- 12909645 TI - Evidence for an intracellular ADP-ribosyl cyclase/NAD+-glycohydrolase in brain from CD38-deficient mice. AB - Cyclic ADP-ribose, a metabolite of NAD+, is known to modulate intracellular calcium levels and signaling in various cell types, including neural cells. The enzymes responsible for producing cyclic ADP-ribose in the cytoplasm of mammalian cells remain unknown; however, two mammalian enzymes that are capable of producing cyclic ADP-ribose extracellularly have been identified, CD38 and CD157. The present study investigated whether an ADP-ribosyl cyclase/NAD+-glycohydrolase independent of CD38 is present in brain tissue. To address this question, NAD+ metabolizing activities were accurately examined in developing and adult Cd38-/- mouse brain protein extracts and cells. Low ADP-ribosyl cyclase and NAD+ glycohydrolase activities (in the range of pmol of product formed/mg of protein/min) were detected in Cd38-/- brain at all developmental stages studied. Both activities were found to be associated with cell membranes. The activities were significantly higher in Triton X-100-treated neural cells compared with intact cells, suggesting an intracellular location of the novel cyclase. The cyclase and glycohydrolase activities were optimal at pH 6.0 and were inhibited by zinc, properties which are distinct from those of CD157. Both activities were enhanced by guanosine 5'-O-(3-thiotriphosphate), a result suggesting that the novel enzyme may be regulated by a G protein-dependent mechanism. Altogether our results indicate the presence of an intracellular membrane-bound ADP-ribosyl cyclase/NAD+-glycohydrolase distinct from CD38 and from CD157 in mouse brain. This novel enzyme, which is more active in the developing brain than in the adult tissue, may play an important role in cyclic ADP-ribose-mediated calcium signaling during brain development as well as in adult tissue. PMID- 12909646 TI - The stereospecific hydroxylation of [2,2-2H2]butane and chiral dideuteriobutanes by the particulate methane monooxygenase from Methylococcus capsulatus (Bath). AB - Experiments on cryptically chiral ethanes have indicated that the particulate methane monooxygenase (pMMO) from Methylococcus capsulatus (Bath) catalyzes the hydroxylation of ethane with total retention of configuration at the carbon center attacked. This result would seem to rule out a radical mechanism for the hydroxylation chemistry, at least as mediated by this enzyme. The interpretation of subsequent experiments on n-propane, n-butane, and n-pentane has been complicated by hydroxylation at both the pro-R and pro-S secondary C-H bonds, where the hydroxylation takes place. It has been suggested that these results merely reflect presentation of both the pro-R and pro-S C-H bonds to the hot "oxygen atom" species generated at the active site, and that the oxo-transfer chemistry, in fact, proceeds concertedly with retention of configuration. In the present work, we have augmented these earlier studies with experiments on [2,2 2H2]butane and designed d,l form chiral dideuteriobutanes. Essentially equal amounts of (2R)-[3,3-2H2]butan-2-ol and (2R)-[2-2H1]butan-2-ol are produced upon hydroxylation of [2,2-2H2]butane. The chemistry is stereospecific with full retention of configuration at the secondary carbon oxidized. In the case of the various chiral deuterated butanes, the extent of configurational inversion has been shown to be negligible for all the chiral butanes examined. Thus, the hydroxylation of butane takes place with full retention of configuration in butane as well as in the case of ethane. These results are interpreted in terms of an oxo-transfer mechanism based on side-on singlet oxene insertion across the C-H bond similar to that previously noted for singlet carbene insertion (Kirmse, W., and Ozkir, I. S. (1992) J. Am. Chem. Soc. 114, 7590-7591). Finally, we discuss how even the oxene insertion mechanism, with "spin crossover" in the transition state, could lead to small amounts of radical rearrangement products, if and when such products are observed. A scheme is described that unifies the two extreme mechanistic limits, namely the concerted oxene insertion and the hydrogen abstraction radical rebound mechanism within the same over-arching framework. PMID- 12909647 TI - Pseudomonas aeruginosa alginate is refractory to Th1 immune response and impedes host immune clearance in a mouse model of acute lung infection. AB - Pseudomonas aeruginosa is an opportunistic respiratory pathogen that accounts for most of the morbidity and mortality in cystic fibrosis (CF) patients. In CF affected lungs, the bacteria undergo conversion from a non-mucoid to a non tractable mucoid phenotype, due to overproduction of alginate. The effect of alginate production on pathogenicity was investigated by using an acute lung infection mouse model that compared a non-mucoid P. aeruginosa strain, PAO1, to its constitutive alginate-overproducing derivative, Alg(+) PAOmucA22, and an alginate-defective strain, Alg(-) PAOalgD. Bacterial suspensions were instilled into the left bronchus and examined 24 and 48 h post-infection. The highest bacterial loads and the most severe lung pathology were observed with strain Alg( ) PAOalgD at 24 h post-infection, which may have been due to an increase in expression of bacterial elastase by the mutant. Significantly lower lung and spleen bacterial loads were found in the two non-mucoid (PAO1 and Alg(-) PAOalgD) groups, compared to the mucoid Alg(+) PAOmucA22 group, between 24 and 48 h post infection. The positive correlation between lung bacteriology and lung macroscopic pathology in the Alg(+) PAOmucA22 group suggests that alginate production not only impedes pulmonary clearing, but also results in severe lung damage. Positive correlations between IL12 levels and lung macroscopic pathology, and between IL12 and IFN-gamma levels in the Alg(+) PAOmucA22 group, suggested a possible contribution of these pro-inflammatory cytokines to tissue damage. No significant differences were found between the three groups in lung cytokine responses at 24 or 48 h post-infection. However, on comparison within each group at 24 and 48 h post-infection, a significant increase in the pro-inflammatory cytokine IFN-gamma was observed. Higher ratios of IFN-gamma/IL4 and IFN gamma/IL10, but lower IL10 levels, were also found in all three groups. These results indicate a Th1-predominated immune response in these animals. Such cytokine responses could have aided the clearance of non-mucoid P. aeruginosa, but were not sufficient to alleviate infection by the mucoid variants. Alginate production may promote survival and persistence of this pathogenic micro-organism in the lung. PMID- 12909648 TI - Anaerobiosis-induced virulence of Salmonella enterica subsp. enterica serovar Typhimurium: role of phospholipase Cgamma signalling cascade. AB - Salmonella enterica subsp. enterica serovar Typhimurium (S. Typhimurium) can initiate entry into non-phagocytic epithelial cells by triggering certain signal transduction pathways, thereby allowing the pathogen to invade and establish a niche within host cells. Anaerobiosis has been shown to be an important inducer of the invasion process of S. Typhimurium. However, the effect of anaerobiosis on modulation of cell signalling cascades by S. Typhimurium is not known. In the present study, the phospholipase Cgamma signalling cascade was investigated in mice enterocytes, following interaction with S. Typhimurium grown under aerobic and anaerobic growth conditions. Significant increases in enterocyte intracellular calcium and inositol 1,4,5-triphosphate levels were observed on interaction with S. Typhimurium grown anaerobically compared with S. Typhimurium grown aerobically. An increased membrane/cytosolic ratio of protein kinase C was also seen with anaerobic S. Typhimurium in enterocytes compared with aerobic S. Typhimurium. These data suggest that anaerobically grown organisms are more efficient in initiating cell-signalling events than are aerobically grown bacteria. These enhanced cell signals may contribute to the increased virulence of S. Typhimurium grown anaerobically. PMID- 12909649 TI - Characterization of biofilm formation by clinical isolates of Mycobacterium avium. AB - Mycobacterium avium is an environmental organism encountered in natural and urban water sources as well as soil. M. avium biofilm has recently been identified on sauna walls and in city water pipes and might have a role in the survival of virulent strains in the environment and in the host. To characterize the M. avium biofilm, an in vitro model was adapted wherein biofilm develops on a PVC surface. Biofilm was detected by staining with crystal violet and visualization by optical microscopy and quantified by A(570). M. avium strains MAC 101, MAC 100, MAC 104, MAC 109, MAC A5 and MAC 5501 (all isolated from the blood of AIDS patients) were used in the assays. Biofilm formation was dependent on the presence of Ca(2+), Mg(2+) or Zn(2+) ions in the water, with the maximal effect seen at a concentration of 1 micro M. The presence of 2 % glucose and peptone as sources of carbon increased the formation of biofilm, while this was partially inhibited by humic acid. Since sliding motility has been associated with the amount of glycopeptidolipid (GPL), TLC was used to determine the presence of GPL. The supernatant of a biofilm-forming culture induced formation of a stable biofilm and amikacin blocked the establishment of biofilm by M. avium strains at subinhibitory concentrations. Bacteria in the biofilm were more resistant to chlorine as well as to exposure to potassium monopersulfate and chloroheximide acetate than were planktonic bacteria. Identification of M. avium genes involved in biofilm formation and further studies of the effect of antimicrobials on the establishment of biofilm may identify approaches for inhibiting M. avium biofilm formation and colonization. PMID- 12909650 TI - Detection of Clostridium difficile cytotoxin and Clostridium perfringens enterotoxin in cases of diarrhoea in the community. AB - Faecal specimens from 843 cases of diarrhoea in the community were tested for the presence of Clostridium difficile cytotoxin and Clostridium perfringens enterotoxin. C. difficile cytotoxin was detected in faecal specimens from 0.6 % of cases aged at least 2 years by using a Vero cell assay. Factors associated with detection of C. difficile cytotoxin were antibiotic therapy, age over 60 years and living in a home with other elderly people. Three methods were used for the detection of C. perfringens enterotoxin: a Vero cell assay, a commercial (TechLab) enzyme immunoassay (EIA) and an in-house EIA. The lower level of detection of pure C. perfringens enterotoxin in buffer was 0.01 micro g ml(-1) by the TechLab EIA and 1.0 micro g ml(-1) by the Vero cell assay. C. perfringens enterotoxin was detected by using the TechLab EIA in faecal specimens from 2.5 % of cases. This commercial EIA was less sensitive than the in-house EIA, detecting only 31 % of positive cases, but was specific and could be used for outbreak investigation by routine diagnostic laboratories. Age over 60 years was a factor associated with C. perfringens enterotoxin detection; this age group may be targeted for testing. PMID- 12909651 TI - Development of a diagnostic PCR assay that targets a heat-shock protein gene (groES) for detection of Pseudomonas spp. in cystic fibrosis patients. AB - Laboratory detection of Pseudomonas spp., in particular Pseudomonas aeruginosa, remains an important assay in the management of patients with cystic fibrosis (CF). As the groES and groEL genes of P. aeruginosa have now been cloned and their nucleotide sequences determined, the aim of this study was to develop a novel PCR assay for the detection of Pseudomonas spp. from patients with CF by employing conserved primer regions of the groE heat-shock protein domain gene. A PCR assay was designed that targeted a 536 bp region of the groE gene to detect Pseudomonas spp. PCR amplification of genomic DNA from extracted organisms generated an amplicon of the expected size (approx. 536 bp) for all P. aeruginosa (n = 60), Pseudomonas putida, Pseudomonas fluorescens and Pseudomonas stutzeri isolates examined, but did not produce a positive amplicon for several other genera and species that are commonly isolated from the sputum of CF patients. RFLP analysis of the amplicons of all P. aeruginosa isolates demonstrated a single RFLP type that consisted of three bands at approximately 80, 190 and 250 bp; direct sequencing of the amplicons demonstrated the presence of a single sequence type, indicating the highly conserved nature of this region. In addition, the assay successfully produced a positive signal from primary non selective plates of three known P. aeruginosa culture-positive CF patients, but was unable to generate a signal in a further six CF patients who had no history of infection with P. aeruginosa or other Pseudomonas spp. This assay is recommended to detect the presence of Pseudomonas spp., including P. aeruginosa, from primary culture plates that originate from laboratory analysis of CF patients' sputum, particularly at review, in those patients with no previous history of Pseudomonas infection or those who appear to be transiently colonized by this organism. Employment of such molecular methodologies, in conjunction with routine clinical sputum cultures, may provide improved information on the microbial status of CF patients, which will aid clinicians in both optimum patient management in terms of antibiotic regimes and CF centre infection-control practices. PMID- 12909652 TI - Assessment of PCR-DGGE for the identification of diverse Helicobacter species, and application to faecal samples from zoo animals to determine Helicobacter prevalence. AB - Helicobacter species are fastidious bacterial pathogens that are difficult to culture by standard methods. A PCR-denaturing gradient gel electrophoresis (PCR DGGE) technique for detection and identification of different Helicobacter species was developed and evaluated. The method involves PCR detection of Helicobacter DNA by genus-specific primers that target 16S rDNA and subsequent differentiation of Helicobacter PCR products by use of DGGE. Strains are identified by comparing mobilities of unknown samples to those determined for reference strains; sequence analysis can also be performed on purified amplicons. Sixteen DGGE profiles were derived from 44 type and reference strains of 20 Helicobacter species, indicating the potential of this approach for resolving infection of a single host by multiple Helicobacter species. Some more highly related species were not differentiated whereas in highly heterogeneous species, sequence divergence was observed and more than one PCR-DGGE profile was obtained. Application of the PCR-DGGE method to DNA extracted from faeces of zoo animals revealed the presence of Helicobacter DNA in 13 of 16 samples; a correlation was seen between the mobility of PCR products in DGGE analysis and DNA sequencing. In combination, this indicated that zoo animals are colonized by a wide range of different Helicobacter species; seven animals appeared to be colonized by multiple Helicobacter species. By this approach, presumptive identifications were made of Helicobacter bilis and Helicobacter hepaticus in a Nile crocodile, Helicobacter cinaedi in a baboon and a red panda, and Helicobacter felis in a wolf and a Taiwan beauty snake. All of these PCR products ( approximately 400 bp) showed 100 % sequence similarity to 16S rDNA sequences of the mentioned species. These results demonstrate the potential of PCR-DGGE-based analysis for identification of Helicobacter species in complex ecosystems, such as the gastrointestinal tract, and could contribute to a better understanding of the ecology of helicobacters and other pathogens with a complex aetiology. PMID- 12909653 TI - Simple and rapid detection of Salmonella strains by direct PCR amplification of the hilA gene. AB - The suitability of a PCR procedure using a pair of primers targeting the hilA gene was evaluated as a means of detecting Salmonella species. A total of 33 Salmonella strains from 27 serovars and 15 non-Salmonella strains from eight different genera were included. PCR with all the Salmonella strains produced a 784 bp DNA fragment that was absent from all the non-Salmonella strains tested. The detection limit of the PCR was 100 pg with genomic DNA and 3 x 10(4) c.f.u. ml(-1) with serial dilutions of bacterial culture. An enrichment-PCR method was further developed to test the sensitivity of the hilA primers for the detection of Salmonella in faecal samples spiked with different concentrations of Salmonella choleraesuis subsp. choleraesuis serovar Typhimurium. The method described allowed the detection of Salmonella Typhimurium in faecal samples at a concentration of 3 x 10(2) c.f.u. ml(-1). In conclusion, the hilA primers are specific for Salmonella species and the PCR method presented may be suitable for the detection of Salmonella in faeces. PMID- 12909654 TI - Assessment of Chlamydia trachomatis infection of semen specimens by ligase chain reaction. AB - Diagnostic potential of the Chlamydia trachomatis ligase chain reaction system (LCx) to assess the presence of C. trachomatis in urine and semen specimens was evaluated. Paired urine and semen specimens from 153 asymptomatic male partners of subfertile couples attending our Center for Reproductive Medicine were examined by LCx. As controls, 19 semen samples from four donors who were participating in the programme for artificial insemination were used. Of these, 12 samples had previously been shown to be C. trachomatis-positive by an in-house PCR. C. trachomatis was detected by LCx in seven of 153 (5 %) urine samples. None of the 153 semen samples tested positive by LCx. Also, none of the 12 C. trachomatis-containing control semen samples were positive by LCx. By in-house PCR, seven urine specimens and two of 153 (1 %) semen samples tested positive. The corresponding urine samples of these male partners were also C. trachomatis positive, as well as the 12 C. trachomatis-containing samples from donors. In conclusion, LCx is not sensitive enough to assess the presence of C. trachomatis in semen specimens; therefore, this method is not recommended to routinely screen semen specimens from donors who participate in programmes for artificial insemination or male partners of subfertile couples for C. trachomatis. PMID- 12909655 TI - Multilocus restriction typing: a tool for Neisseria meningitidis strain discrimination. AB - Invasive disease-associated strains of Neisseria meningitidis were analysed by multilocus restriction typing (MLRT), which involves the restriction fragment length polymorphism analysis of PCR products generated from the seven loci of housekeeping genes used in MLST. Several different restriction patterns (alleles) were observed for each of the seven loci examined. Greater allelic variation was observed with the fumC and pgm loci than with the abcZ and adk loci, suggesting that the latter were more conserved. The alleles at each of the seven loci were combined to give an allelic profile or restriction type (RT). A good correlation between RT and serogroup, serotype and serosubtype was observed, as all C 2ap1.2,5 strains were contained in a single RT, as were all but one strain of B 4p1.4. In this study, MLRT proved to be an efficient, effective and relatively inexpensive method for N. meningitidis strain characterization. PMID- 12909656 TI - Human rhabdomyosarcoma cells for rapid detection of enteroviruses by shell-vial assay. AB - The ability of the RD (rhabdomyosarcoma) and MRC-5 cell-lines to detect enteroviruses in 33 clinical samples (cerebrospinal fluid, stools and throat swabs) was evaluated. The samples had previously tested enterovirus-positive by traditional tube-culture and had been frozen after their initial processing. By traditional tube-culture, 100 and 85 % of samples were positive for enterovirus in RD and MRC-5 cells, respectively. By rapid shell-vial assay, 94 and 45.5 % were positive after 48 h incubation in RD and MRC-5 cells, respectively. RD cells supported growth of all enterovirus serotypes, whereas MRC-5 cells were not able to detect any of the three coxsackieviruses that were found (one coxsackievirus A9 and two coxsackievirus B5). The shell-vial assay with RD cell-lines may be a useful tool for rapid diagnosis of enteroviral infection. PMID- 12909657 TI - Direct-test PCR for detection of meningococcal DNA and its serogroup characterization: standardization and adaptation for use in a public health laboratory. AB - A direct PCR test (DT-PCR) was established to detect Neisseria meningitidis DNA in clinical samples from patients with suspected bacterial meningitis. Specific primers for the 16S rDNA of N. meningitidis were designed to amplify a 600 bp DNA fragment. One hundred and ninety-three clinical samples were analysed, corresponding to 114 samples from patients diagnosed as positive and 79 as negative for infection by N. meningitidis using conventional methods (culture, latex agglutination and counterimmunoelectrophoresis). These samples were submitted to PCR by two different clinical sample preparation approaches (with and without DNA extraction and purification) and submitted to different PCR protocols to improve the results. In agarose gel detection, the sensitivity value for DT-PCR was 88.5 % and, using dot-blot DNA detection, the sensitivity increased to 96.4 %. The detection limit for meningococcus in cerebrospinal fluid was 2x10(2) c.f.u. ml(-1). Serogroup prediction was done using a multiplex PCR protocol and the sensitivity was 83 % for agarose gel DNA detection and 96.4 % using dot-blot DNA detection. PMID- 12909658 TI - Relationship between molecular epidemiology and antibiotic susceptibility of methicillin-resistant Staphylococcus aureus (MRSA) in a French teaching hospital. AB - The objective of this study was to investigate the relationship between molecular epidemiology and antibiotic susceptibility of methicillin-resistant Staphylococcus aureus (MRSA) over a period of 4 years. The antibiotype of all MRSA isolates that were identified during a yearly period of 3 months was determined; 50 consecutive non-replicate MRSA isolates were typed each year. Susceptibility rates to gentamicin, tobramycin and ofloxacin remained stable (95, 16 and 4 %, respectively). In contrast, the proportion of MRSA isolates susceptible to erythromycin increased progressively from 10.5 to 32.5 % (P < 0.001). PFGE analysis of genomic DNA from 200 isolates revealed the presence of 15 different clones. Two epidemic clones were identified, which contained 150 (clone A) and 28 (clone C) isolates. Non-epidemic strains were more frequently susceptible to ofloxacin (31.8 versus 1.1 %) and tobramycin (45.4 versus 16.8 %) than epidemic strains; those isolates that were susceptible to all antibiotics tested belonged to sporadic clones. The increase of erythromycin susceptibility within MRSA isolates was caused by the emergence of clone C. This study suggests that when selection pressure exerted by an antibiotic is insufficient (i.e. below a threshold level), fitness advantages play a predominant role in the dissemination of MRSA clones. The balance between the selection pressure exerted by antibiotics and the disadvantage of lower replication rates of resistant strains in the absence of antibiotics complicates the biological model of clonal dissemination of epidemic MRSA strains. PMID- 12909659 TI - First isolation and characterization of Borrelia burgdorferi sensu lato strains from Ixodes ricinus ticks in Turkey. AB - In order to investigate the presence and prevalence of Lyme borreliosis (Lyme disease) Borrelia species, 312 unfed ticks were collected by flagging at a woodland area in Trakya, in the European side of Turkey, in May 2002. Twelve of 299 Ixodes ricinus ticks were infected with Borrelia spp., as determined by cultivation in BSK medium (prevalence rate 4.0 %). Ten pure cultures were subjected to further characterization by sequencing analysis of the 5S-23S rDNA intergenic spacer, 16S rDNA and flagellin gene. One isolate of Borrelia burgdorferi sensu stricto, two of Borrelia garinii (Eurasian type), two of Borrelia afzelii, four of Borrelia lusitaniae and one of Borrelia valaisiana were identified. However, no Asian-type B. garinii was found. Interestingly, all Borrelia species that are known to be carried by I. ricinus were discovered among the 10 isolates. These results provide the first evidence for the existence of the Lyme borreliosis agent in Turkey. PMID- 12909660 TI - Changes in genetic types and population dynamics of Moraxella catarrhalis in hospitalized children are not associated with an exacerbation of existing disease. AB - Pulsed-field gel electrophoresis typing was performed on a retrospective set of 129 Moraxella catarrhalis isolates obtained over a 20 month period from 70 children admitted to, or presenting at, the Erasmus University Medical Center, Rotterdam, The Netherlands. The mean age of the children (at the end of the study) was 2.5 years, with a range of 6 months to 15 years. Fifty-one different M. catarrhalis types were isolated from the hospitalized children, with 31 % (22/70) being infected with two particularly prevalent M. catarrhalis types. These two prevalent types also exhibited different protein profiles. The majority (72%; 16/22) of the children infected with these two predominant types had spent at least 1 week on two paediatric intensive care wards. No exacerbation of existing disease or new disease was observed in children who experienced M. catarrhalis type changes. PMID- 12909661 TI - Molecular characterization of clinical and environmental isolates of vancomycin resistant Enterococcus faecium and Enterococcus faecalis from a teaching hospital in Wales. AB - The present study describes the first molecular characterization of environmental and clinical isolates of vancomycin-resistant enterococci (VRE) in Wales. Over a 3-month period (May-July 2000), 134 isolates of VRE (89 Enterococcus faecium and 45 Enterococcus faecalis) were isolated from the patient environment of the University Hospital of Wales (UHW) in Cardiff, Wales, UK. In addition, over the same time-period, 24 clinical isolates of VRE (20 isolates of E. faecium and four isolates of E. faecalis) were obtained from 14 patients. All study isolates were subjected to PFGE typing and their van genotypes were determined by using multiplex PCR. The vanA PCR product (231 bp) was evident in 146 (92 %) of 158 VRE isolates; the remaining 12 isolates (8 %) were positive for the vanB gene. All isolates of E. faecalis were found to be vanA-positive. In total, 16 PFGE banding profiles (pulsotypes) were observed for environmental isolates of E. faecium, whilst eight pulsotypes were found for isolates of E. faecalis. Some of these pulsotypes were isolated from multiple sites, whereas others were more restricted in their distribution. Eleven pulsotypes were evident for clinical isolates and eight of these (representing 11 isolates) were also encountered in environmental isolates. Eleven clinical isolates of E. faecium (55 %) shared an identical pulsotype that was not detected in environmental isolates. These results demonstrate a heterogeneous environmental population of VRE and an association of certain strains with clinical isolates. Predominance of a single pulsotype (not detected in the environment) amongst clinical isolates suggests non-environmental transmission between patients. PMID- 12909662 TI - Bacteriocin-like inhibitory substance (BLIS) production by the normal flora of the nasopharynx: potential to protect against otitis media? AB - The normal bacterial flora of the upper airways provides an important barrier to invading pathogens. This study investigated the production of bacteriocin-like inhibitory substances (BLIS) by streptococci isolated from the nasopharyngeal flora of children who either do or do not experience recurrent acute otitis media (AOM). Twenty children with recurrent AOM and 15 controls were tested. Swabs from the nasopharynx were evaluated for streptococci having BLIS activity against two representative strains of each of the AOM pathogens Streptococcus pneumoniae, Streptococcus pyogenes, Haemophilus influenzae and Moraxella catarrhalis. Streptococci displaying strong BLIS activity were characterized further and tested for known streptococcal bacteriocin structural genes. Sixty-five per cent of children had nasopharyngeal streptococcal isolates that were inhibitory to strains of one or more of the AOM pathogens. Six children (17 %) had streptococci that demonstrated strong BLIS activity against strains of at least three of the pathogenic species. Three of these inhibitory isolates were Streptococcus salivarius, two were S. pneumoniae and one was S. pyogenes. The inhibitory S. salivarius and S. pyogenes were shown to have structural genes for known streptococcal bacteriocins. No statistically significant difference was found between the two groups of children with respect to the presence of inhibitory streptococci in their nasopharyngeal floras. The finding of S. salivarius with strong inhibitory activity against several AOM pathogens in the nasopharyngeal flora of children is unique. Although there is no clear evidence from the present study that these organisms protect against AOM, their low pathogenicity and strong in-vitro BLIS production capability indicate that they should be incorporated in future trials of bacteriotherapy for recurrent AOM. PMID- 12909663 TI - Fatal disseminated Acremonium strictum infection in a preterm newborn: a very rare cause of neonatal septicaemia. AB - Species of the genus Acremonium (Cephalosporium) are opportunistic micro organisms that are environmentally widespread saprophytes in soil and can, very rarely, be pathogenic in humans. Disseminated infection has been described in patients with immunodeficiency, but has previously been reported in only one neonate. A preterm infant with Acremonium strictum fungaemia is reported here. The patient was born at 27 weeks gestation and weighed 870 g at birth. She needed intensive respiratory management and became septic on day 11 of life. Blood and cerebrospinal fluid (CSF) cultures were positive for A. strictum. The patient did not respond to therapy with amphotericin B plus fluconazole and died on day 25 of life. The autopsy showed foci due to A. strictum in the brain, liver and heart. PMID- 12909664 TI - Characterization of a novicida-like subspecies of Francisella tularensis isolated in Australia. AB - Francisella tularensis is found throughout the Northern Hemisphere, where it is associated with the disease of tularaemia in animals and humans. The isolation and identification is reported of a novicida-like subspecies of F. tularensis from a foot wound sustained in brackish water in the Northern Territory of Australia. PMID- 12909665 TI - Plant-based siderophore: a new avenue in molecular medicine for tuberculosis. PMID- 12909668 TI - Human muscle protein synthesis is modulated by extracellular, not intramuscular amino acid availability: a dose-response study. AB - To test the hypothesis that muscle protein synthesis (MPS) is regulated by the concentration of extracellular amino acids, we investigated the dose-response relationship between the rate of human MPS and the concentrations of blood and intramuscular amino acids. We increased blood mixed amino acid concentrations by up to 240 % above basal levels by infusion of mixed amino acids (Aminosyn 15, 44 261 mg kg-1 h-1) in 21 healthy subjects, (11 men 10 women, aged 29 +/- 2 years) and measured the rate of incorporation of D5-phenylalanine or D3-leucine into muscle protein and blood and intramuscular amino acid concentrations. The relationship between the fold increase in MPS and blood essential amino acid concentration ([EAA], mM) was hyperbolic and fitted the equation MPS = (2.68 x [EAA])/(1.51 + [EAA]) (P < 0.01). The pattern of stimulation of myofibrillar, sarcoplasmic and mitochondrial protein was similar. There was no clear relationship between the rate of MPS and the concentration of intramuscular EAAs; indeed, when MPS was increasing most rapidly, the concentration of intramuscular EAAs was below basal levels. We conclude that the rates of synthesis of all classes of muscle proteins are acutely regulated by the blood [EAA] over their normal diurnal range, but become saturated at high concentrations. We propose that the stimulation of protein synthesis depends on the sensing of the concentration of extracellular, rather than intramuscular EAAs. PMID- 12909675 TI - 5-Hydroxytryptamine 1A receptors inhibit cold-induced sympathetically mediated cutaneous vasoconstriction in rabbits. AB - 5-HT1A receptor agonists lower body temperature. We have investigated whether activation of 5-HT1A receptors inhibits cutaneous sympathetic discharge so that dilatation of the cutaneous vascular bed lowers body temperature by increasing heat transfer to the environment. We measured ear pinna blood flow in conscious rabbits (with chronically implanted Doppler ultrasound flow probes), and postganglionic sympathetic vasomotor nerve activity in anaesthetized rabbits. Recordings from conscious rabbits were made in a cage at 26 degrees C and the rabbit was then transferred to a cage at 10 degrees C. The ear pinna Doppler signal fell from 56 +/- 4 cm s-1 in the 26 degrees C cage to 4 +/- 1 cm s-1 (P < 0.0001, n = 24) after 30 min in the 10 degrees C cage, and body temperature increased from 38.8 +/- 0.2 to 39.0 +/- 0.2 degrees C (P < 0.01, n = 24). The 5 HT1A agonist 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT; 0.1 mg kg-1 I.V.) reversed the cold-induced fall in ear pinna blood flow (Doppler signal increased from 5 +/- 1 to 55 +/- 8 cm s-1, P < 0.001, n = 7) within 5 min when administered 30 min after transfer to the 10 degrees C cage, and prevented the fall in ear pinna blood flow when administered before the rabbit was transferred to the 10 degrees C cage. Body temperature decreased after administration of 8-OH-DPAT. These changes were abolished by the specific 5-HT1A antagonist WAY-100635 (0.1 mg kg-1 I.V.). In anaesthetized rabbits, 8-OH-DPAT (0.1 mg kg-1 I.V.) reduced resting postganglionic cutaneous sympathetic vasomotor discharge, and prevented the increase normally elicited by cooling the trunk. Our experiments constitute the first demonstration that activation of 5-HT1A receptors powerfully inhibits cold-induced increases in cutaneous sympathetic vasomotor discharge, thereby dilating the cutaneous vascular bed and increasing transfer of heat to the environment. PMID- 12909676 TI - Mg2+ dependence of halothane-induced Ca2+ release from the sarcoplasmic reticulum in rat skeletal muscle. AB - The effect of cytosolic Mg2+ on halothane-induced Ca2+ release from the sarcoplasmic reticulum (SR) was investigated in mechanically skinned fibres from the rat extensor digitorum longus (EDL) muscle. Preparations were perfused with solutions mimicking the intracellular milieu and changes in [Ca2+] were detected using Fura-2 fluorescence. In the presence of 1 mM Mg2+, brief (500 ms) applications of 40 mM halothane failed to induce Ca2+ release from the SR. However, Ca2+ release became detectable when [Mg2+] was reduced to 0.4 mM, and the amplitude of the response increased progressively as [Mg2+] was further reduced to 0.2 and 0.1 mM. Lower halothane concentrations within the range found during anaesthesia or induction (0.1-1.2 mM) failed to induce SR Ca2+ release at 0.2 or 0.4 mM Mg2+. However, in further experiments, preparations were exposed to 1 mM halothane for 2-3 min under conditions where the volume of solution surrounding the preparation was restricted by stopping the flow. In the absence of perfusion, 1 mM halothane induced Ca2+ release from the SR at 0.4 mM Mg2+ in two out of six preparations, and release was observed consistently at 0.2 and 0.1 mM Mg2+. Responses to 1 mM halothane induced in the presence of 0.4 and 0.2 mM Mg2+ were typically delayed in onset and involved a localised release of Ca2+ that propagated along the fibre. These results suggest that halothane-induced Ca2+ release is strongly inhibited at normal physiological levels of Mg2+. However, when Mg2+-induced inhibition of the ryanodine receptor (RYR) is reduced, levels of halothane within the range found during anaesthesia can induce a marked efflux of Ca2+ from the SR. This may be of relevance to the condition of malignant hyperthermia, where the inhibition of RYRs by Mg2+ is reportedly reduced. PMID- 12909679 TI - Spinal and supraspinal functions of noradrenaline in the frog embryo: consequences for motor behaviour. AB - The monoamine noradrenaline (NA) can initiate and/or modulate locomotion in a variety of vertebrates. Here we report that exogenous NA application can facilitate two completely different fictive behaviours in embryos of the common frog Rana temporaria, depending on whether spinal networks are connected to supraspinal centres. When the nervous system is intact, NA elicits a non-rhythmic coiling motor response, reminiscent of a spontaneous behaviour appropriate to drive hatching movements, but has only minor effects on evoked swimming activity. After the spinal cord has been severed from the brain, spontaneous coiling is no longer observed, nor can NA elicit it, but the amine can 'release' swimming rhythm generation in response to electrical skin stimulation. The rhythm is similar, but relatively inflexible when compared to fictive swimming recorded from intact animals. Our pharmacological tests indicate that alpha 1 adrenoreceptors are involved in the permissive role of NA during spinalised rhythmic swimming and that the fictive coiling response to NA in intact animals involves descending inputs and the activation of beta 1-adrenoreceptors. Furthermore, the subtle effects of NA on evoked swimming in intact animals were mimicked by either alpha 1- or alpha 2-adrenoreceptor activation, reversibly decreasing motor burst durations and increasing their frequency. We discuss our results with reference to the known synergistic actions of NA with another aminergic neuromodulator, serotonin, and raise the possibility that these amines may actively regulate the release of one another during locomotion, in addition to their respective post-synaptic targets in the spinal cord. PMID- 12909680 TI - Kainate receptor (GluR5)-mediated disinhibition of responses in rat ventrobasal thalamus allows a novel sensory processing mechanism. AB - Kainate receptors have been studied extensively in vitro, but how they might function physiologically remains unclear. We studied kainate receptor modulation of synaptic responses in the rat ventrobasal thalamus using the novel antagonist LY382884 and the agonist ATPA (selective for GluR5-containing kainate receptors) as tools. No evidence could be found for a direct contribution of kainate receptors to responses of thalamic relay cells to lemniscal (sensory) input in thalamic slices studied with the aid of intracellular and field potential recordings, using selective AMPA and NMDA receptor antagonists and LY382884. However, the GluR5 agonist ATPA reduced the IPSPs originating from the thalamic reticular nucleus. Extracellular single-neurone recordings in anaesthetised rats showed that excitatory responses evoked by physiological vibrissa afferent stimulation were reduced by LY382884 applied iontophoretically at the recording site. This action of the antagonist was occluded when GABA receptors were blocked, indicating that the reduction in excitatory sensory responses by LY382884 is due to an action on GABAergic inhibition arising from the thalamic reticular nucleus. Further experiments showed that these actions depended on whether inhibition was evoked during activation of the excitatory receptive field rather than when inhibition was evoked from a surround vibrissa. We suggest that GluR5 is located presynaptically on inhibitory GABAergic terminals of thalamic reticular nucleus neurones, and that it is normally activated by glutamate spillover from synapses between excitatory afferents and relay neurones during physiological stimulation. We propose that this GluR5-activated disinhibition has an important novel role in extracting sensory information from background noise. PMID- 12909682 TI - Measurement of voluntary activation of fresh and fatigued human muscles using transcranial magnetic stimulation. AB - Recently, transcranial magnetic stimulation of the motor cortex (TMS) revealed impaired voluntary activation of muscles during maximal efforts. Hence, we evaluated its use as a measure of voluntary activation over a range of contraction strengths in both fresh and fatigued muscles, and compared it with standard twitch interpolation using nerve stimulation. Subjects contracted the elbow flexors isometrically while force and EMG from biceps and triceps were recorded. In one study, eight subjects made submaximal and maximal test contractions with rests to minimise fatigue. In the second study, eight subjects made sustained maximal contractions to reduce force to 60 % of the initial value, followed by brief test contractions. Force responses were recorded following TMS or electrical stimulation of the biceps motor nerve. In other contractions, EMG responses to TMS (motor evoked potentials, MEPs) or to stimulation at the brachial plexus (maximal M waves, Mmax) were recorded. During contractions of 50 % maximum, TMS elicited large MEPs in biceps (> 90 % Mmax) which decreased in size (to approximately 70 % Mmax) with maximal efforts. This suggests that faster firing rates made some motor units effectively refractory. With fatigue, MEPs were also smaller but remained > 70 % Mmax for contractions of 50-100 % maximum. For fresh and fatigued muscle, the superimposed twitch evoked by motor nerve and motor cortex stimulation decreased with increasing contraction strength. For nerve stimulation the relation was curvilinear, and for TMS it was linear for contractions of 50-100 % maximum (r2 = 1.00). Voluntary activation was derived using the expression: (1 - superimposed twitch/resting twitch) x 100. The resting twitch was measured directly for nerve stimulation and for TMS, it was estimated by extrapolation of the linear regression between the twitch and voluntary force. For cortical stimulation, this resulted in a highly linear relation between voluntary activation and force. Furthermore, the estimated activation corresponded well with contraction strength. Using TMS or nerve stimulation, voluntary activation was high during maximal efforts of fresh muscle. With fatigue, both measures revealed reduced voluntary activation (i.e. central fatigue) during maximal efforts. Measured with TMS, this central fatigue accounted for one-quarter of the fall in maximal voluntary force. We conclude that TMS can quantify voluntary activation for fresh or fatigued muscles at forces of 50-100 % maximum. Unlike standard twitch interpolation of the elbow flexors, voluntary activation measured with TMS varies in proportion to voluntary force, it reveals when extra output is available from the motor cortex to increase force, and it elicits force from all relevant synergist muscles. PMID- 12909683 TI - Human skeletal muscle sympathetic nerve activity, heart rate and limb haemodynamics with reduced blood oxygenation and exercise. AB - Acute systemic hypoxia causes significant increases in human skeletal muscle sympathetic nerve activity (MSNA), heart rate and ventilation. This phenomenon is thought to be primarily mediated by excitation of peripheral chemoreceptors sensing a fall in arterial free oxygen partial pressure (Pa,O2). We directly tested the role of Pa,O2 on MSNA (peroneal microneurography), heart rate, ventilation and leg haemodynamics (n = 7-8) at rest and during rhythmic handgrip exercise by using carbon monoxide (CO) to mimic the effect of systemic hypoxia on arterial oxyhaemoglobin (approximately 20 % lower O2Hba), while normalising or increasing Pa,O2 (range 40-620 mmHg). The four experimental conditions were: (1) normoxia (Pa,O2 approximately 110 mmHg; carboxyhaemoglobin (COHb) approximately 2 %); (2) hypoxia (Pa,O2 approximately 40 mmHg; COHb approximately 2 %); (3) CO + normoxia (Pa,O2 approximately 110 mmHg; COHb approximately 23 %); and (4) CO + hyperoxia (Pa,O2 approximately 620 mmHg; COHb ~24 %). Acute hypoxia augmented sympathetic burst frequency, integrated MSNA, heart rate and ventilation compared to normoxia over the entire protocol (7-13 bursts min-1, 100-118 %, 13-17 beats min-1, 2-4 l min-1, respectively, P < 0.05). The major new findings were: (1) CO + normoxia and CO + hyperoxia also elevated MSNA compared to normoxia (63-144 % increase in integrated MSNA; P < 0.05) but they did not increase heart rate (62 67 beats min-1) or ventilation (6.5-6.8 l min-1), and (2) despite the 4-fold elevation in MSNA with hypoxaemia and exercise, resting leg blood flow, vascular conductance and O2 uptake remained unchanged. In conclusion, the present results suggest that increases in MSNA with CO are not mediated by activation of the chemoreflex, whereas hypoxia-induced tachycardia and hyperventilation are mediated by activation of the chemoreflex in response to the decline in Pa,O2. Our findings also suggest that Pa,O2 is not an obligatory signal involved in the enhanced MSNA with reduced blood oxygenation. PMID- 12909684 TI - A single course of prenatal betamethasone in the rat alters postnatal brain cell proliferation but not apoptosis. AB - The aim of this study was to determine the effects of a clinically relevant single course of prenatal betamethasone in the rat on growth parameters with particular reference to brain cell proliferation and apoptosis. We report that administration of 170 microg kg-1 betamethasone twice within 4 h to E20 pregnant rats conveys moderate somatic growth retardation. Further, using a measure of brain cell proliferation independent of blood-brain barrier (BBB) permeability, we demonstrate for the first time that betamethasone is chronically anti proliferative to brain cells without inducing caspase-3-mediated apoptosis. More importantly we show that there is a significant and sexually divergent rebound of neural proliferation which occurs earlier in males than in females and continues until at least 21 days of postnatal life. BBB permeability to [3H]thymidine was significantly increased by steroid treatment re-iterating the fact that tracer studies not correcting for BBB permeability, such as bromodeoxyuridine (BrdU), may be questionable in this type of study. Further, prenatal steroid treatment did not alter postnatal corticosterone levels. In summary we show that prenatal betamethasone conveys significant and long-lasting side effects and that its human clinical application in preterm labour needs more careful consideration as compared to the relative ease with which it is prescribed today. PMID- 12909686 TI - Capsaicin-sensitive and -insensitive vagal bronchopulmonary C-fibres in the mouse. AB - We developed an isolated tracheally perfused (35-37 degrees C) nerve-lung preparation for the study of bronchopulmonary afferent nerve activity in the mouse. Extracellular recordings were made from the vagal sensory neurons located in the jugular-nodose ganglia complex (JNC) with identified receptive fields in the lungs. Analysis of the vagal compound action potential revealed that the mouse vagal C-fibre conduction velocities range from 0.3 to 1.5 m s(-1). A total of 83 bronchopulmonary C-fibres were studied. The sensitivity of the bronchopulmonary C-fibres to the vanilloid receptor 1 (VR1) agonist capsaicin was dependent on conduction velocity. Thus C-fibres with conduction velocities between 0.3 and 0.7 m s(-1) responded to capsaicin (1 microM) while C-fibres with conduction velocities between 0.7 and 1.5 m s(-1) were capsaicin insensitive. Similarly, bradykinin (1 microM) excited only those C-fibres with conduction velocities < 0.7 m s(-1). The response to bradykinin was not mimicked by the B1 receptor agonist [des-Arg9]bradykinin (1 microM) and was abolished by the bradykinin B2 receptor antagonist HOE 140 (1 microM). Adenosine 5'-triphosphate (ATP, 30 microM) activated the C-fibres irrespective of the conduction velocities. This response was mimicked by the selective P2X agonist alpha,beta methylene-adenosine 5'-triphosphate (30 microM). Consistent with the electrophysiology, morphological analysis revealed that only approximately 40% of the lung-specific small diameter (< 20 microm) JNC neurons consistent with the C fibre cell bodies display VR1 immunoreactivity. This study describes a convenient in vitro method for the study of mouse bronchopulmonary C-fibres. The results indicate that C-fibres in the mouse lungs are not homogeneous, but can be subclassified into capsaicin-sensitive and capsaicin-insensitive phenotypes. PMID- 12909685 TI - GABAA and glycine receptors in regulation of intercostal and abdominal expiratory activity in vitro in neonatal rat. AB - The roles played by GABAA and glycine receptors in inspiratory-expiratory motor co-ordination and in tonic inhibitory regulation of expiratory motor activity were studied using brainstem-spinal cord (-rib) preparations from neonatal rats. Inspiratory activity was recorded from the C4 ventral root. Expiratory activity in internal intercostal muscle, internal oblique muscle or T13 ventral root was evoked by a decrease in perfusate pH from 7.4 to 7.1 (i.e. from normal to low pH conditions) and was limited to the first part of the expiratory phase. Under low pH conditions, bath application of 10 microM bicuculline, a GABAA receptor antagonist, caused the inspiratory burst to overlap the expiratory burst in 2/7 preparations. Overlapping of the expiratory burst with the inspiratory burst was observed in 7/7 preparations made under 10 microM bicuculline. Furthermore, such preparations exhibited expiratory bursts under bicuculline-containing normal pH conditions. Local application of 10 microM bicuculline to the brainstem under normal pH conditions evoked expiratory bursts, some of which overlapped the inspiratory bursts. Picrotoxin, another antagonist of the GABAA receptor, had similar effects. Under normal pH conditions, application of strychnine (0.2- 2.0 microM; a glycine receptor antagonist) to the brainstem did not evoke expiratory bursts. On subsequent application of strychnine-containing low pH solution, expiratory bursts were evoked and some (0.5 microM) or all (2.0 microM) of these overlapped the inspiratory burst. Simultaneous application of picrotoxin and strychnine to the brainstem evoked expiratory bursts that overlapped the inspiratory bursts and a subsequent decrease in perfusate pH to 7.1 increased the frequency of the respiratory rhythm. It was a characteristic finding that the duration of the expiratory burst exceeded that of the inspiratory burst under control low pH conditions. This remained true during concurrent blockade of GABAA and glycine receptors. The results suggest that in the in vitro preparation from neonatal rats: (1) GABAA and glycine receptors within the brainstem play important roles in the co-ordination between inspiratory and expiratory motor activity, (2) tonic inhibition via GABAA receptors, but not glycine receptors, plays a role in the regulation of expiratory motor activity and (3) inspiratory and expiratory burst termination is independent of both GABAA and glycine receptors. PMID- 12909687 TI - Expression analysis of five tobacco EIN3 family members in relation to tissue specific ethylene responses. AB - Ethylene induces different sets of genes in different tissues and at different stages of development. To investigate whether these differential responses are caused by differential expression of members of the EIN3 family transcription factors, five tobacco family members were isolated. They can be divided into three subgroups, which is probably due to the amphidiploid nature of tobacco. In phylogenetic analysis, each of the subgroups clustered with one of the three tomato EIL proteins and all NtEILs proved to be most homologous to Arabidopsis EIN3 and EIL1. Although organ-specific ethylene responses have been observed before, northern blot analysis showed that all NtEILs were expressed in all organs. To study differential NtEIL expression at the cellular level, in situ hybridization was used on the tobacco ovary. It was found that different ovary tissues displayed variable ethylene-induced expression of two ethylene-responsive marker genes. By contrast, no differences were found in expression level or tissue-specificity for any of the NtEILs in the ovary, before or after ethylene treatment. This indicates that the organ and tissue-specific ethylene responses are not caused by differential expression of NtEIL family members. These results support a model in which the developmental signals that regulate the tissue specific responses are integrated with the ethylene signal downstream of a common primary ethylene-signalling pathway. PMID- 12909688 TI - Salt tolerance-related protein STO binds to a Myb transcription factor homologue and confers salt tolerance in Arabidopsis. AB - Regulating the intracellular Na+/K+ ratio is an essential process for salinity tolerance. The yeast mutant, can, which is deficient in calcineurin, can not grow on medium containing Na+ because it is unable to regulate the intracellular Na+/K+ ratio. Expression of the STO gene of Arabidopsis thaliana in the can mutant complements the salt-sensitive phenotype. A protein of Arabidopsis, an H protein promoter binding factor (HPPBF-1), that binds to STO protein was isolated. HPPBF-1 cDNA has a sequence encoding a Myb DNA binding-motif and its gene expression is induced by salt stress. Furthermore, HPPBF-1 protein is localized in the nucleus. Although, the expression level of STO is not induced under salt-stress conditions, overexpression of STO in a transgenic Arabidopsis plant gave it a higher salt tolerance than was observed in the wild type. When STO transgenic plants and wild-type plants were subjected to salt stress, root growth was increased by 33-70% in the transgenic plants under salt stress. These results suggest that STO is involved in salt-stress responses in Arabidopsis. PMID- 12909689 TI - Rate of dehydration of corn (Zea mays L.) pollen in the air. AB - The water content of corn (Zea mays L.) pollen directly affects its dispersal in the atmosphere through its effect on settling speed and viability. Therefore, the rate of water loss from pollen after being shed from the anther is an important component of a model to predict effective pollen transport distances in the atmosphere. The rate of water loss from corn pollen in air was determined using two methods: (1) by direct weighing of samples containing approximately 5 x 10(4) grains, and (2) by microscopic measurement of the change in size of individual grains. The conductance of the pollen wall to water loss was derived from the time rate of change of pollen mass or pollen grain size. The two methods gave average conductance values of 0.026 and 0.027 cm s-1, respectively. In other experiments, the water potential, psi, of corn pollen was determined at various values of relative water content (dry weight basis), either by using a thermocouple psychrometer or by allowing samples of pollen to come to vapour equilibrium with various saturated salt solutions. Non-linear regression analysis of the data yielded psi (MPa) = -3.218 theta(-1.35) (r2 = 0.94; for -298 < or = psi < or = -1 MPa). This result was incorporated into a model differential equation for the rate of water loss from pollen. The model agreed well (r2 approximately 0.98) with the observed time-course of the decrease of water content of pollen grains exposed to a range of temperature and humidity conditions. PMID- 12909690 TI - A method for the isolation of root hairs from the model legume Medicago truncatula. AB - A new method for the isolation of root hairs from the model legume, Medicago truncatula, was developed. The procedure involves the propagation of detached roots on agar plates and the collection of root hairs by immersion in liquid nitrogen. Yields of up to 40 micro g of root hair protein were obtained from 50 100 root tips grown for 3 weeks on a single plate. The high purity of the root hair fraction was monitored by western blot analysis using an antibody to the pea epidermis specific protein PsRH2. Sequence analyses revealed that the protein homologous to PsRH2 in M. truncatula, MtRH2, is identical to the root protein MtPR10-1. The MtRH2 protein proved to be a useful endogenous marker to monitor root hair isolation since it is also specifically expressed in the root epidermis. PMID- 12909691 TI - Cold-specific and light-stimulated expression of a wheat (Triticum aestivum L.) Cor gene Wcor15 encoding a chloroplast-targeted protein. AB - Wcor15, a member of the wheat cold-responsive (Cor) gene family, has been isolated and characterized. The deduced polypeptide WCOR15 (MW=14.7 kDa) showed high homology to the previously identified wheat and barley COR proteins. Southern blot analysis using diploid, tetraploid and hexaploid wheat and diploid Aegilops species showed that the wheat and related wild genomes possessed multiple copies of Wcor15 homologues. Five copies were assigned to the homologous group 2 chromosomes by nulli-tetrasomic analysis. Northern blot analysis showed that expression of Wcor15 was specifically induced by low-temperature. Homologous transcripts accumulated in leaves, and light markedly increased their steady state levels. Bombardment-mediated transient expression analysis of a chimeric CaMV 35S::Wcor15-GFP construct showed protein-targeting to epidermal guard cell chloroplasts in excised spiderwort leaves. A promoter of Wcor15 contained at least three CRT/DRE-like sequence motifs found in Arabidopsis Cor genes and induced the reporter GUS gene expression in leaves of transgenic tobacco plants under low-temperature and light conditions. These results suggest that the functional Cor gene system involving the CRT/DRE cis-element is conserved in both monocotyledonous and dicotyledonous plants. PMID- 12909692 TI - Isolation and characterization of a Brassica napus cDNA corresponding to a B class floral development gene. AB - B-class floral homeotic genes are required for the proper formation and identity of petals and stamens in dicot flowers. A partial cDNA clone encoding a B-class gene, BnAP3 (Brassica napus APETALA3), was isolated from a B. napus cDNA library derived from young inflorescence meristems. The 5' region of the cDNA was retrieved by RACE. The deduced amino acid sequence of the full-length clone exhibited high similarity to APETALA3 of Arabidopsis thaliana and functionally homologous proteins from other species. 5' RACE and Southern analysis suggests that BnAP3 has multiple alleles in B. napus. Expression analysis assayed by RT PCR shows that BnAP3 is expressed in floral tissues, as well as non-floral tissues such as root and bract. Transformation of wild-type A. thaliana and B. napus plants with BnAP3 under the control of a promoter specific to reproductive organs converts carpels to stamens, while the expression of this construct in A. thaliana plants mutant for AP3 restores the development of third-whorl stamens in addition to directing a carpel to stamen conversion in the fourth whorl. PMID- 12909693 TI - Evolution of the cellular stress proteome: from monophyletic origin to ubiquitous function. AB - Cells respond to acute environmental change by activating a stress response that is widely studied. However, knowledge of this stress response is fragmentary, and a unifying concept explaining its universality for many different species and types of stress is lacking. The need for a holistic view emphasizing the key aspects of the stress response is addressed by the following hypothesis. The cellular stress response is a reaction to any form of macromolecular damage that exceeds a set threshold, independent of the underlying cause. It is aimed at temporarily increasing tolerance limits towards macromolecular damage by utilizing a phylogenetically conserved set of genes and pathways that mediate global macromolecular stabilization and repair to promote cellular and organismal integrity under suboptimal conditions. This mechanism affords time for a separate set of stressor-specific adaptations, designed to re-establish cellular homeostasis, to take action. Supporting evidence, emerging conclusions, and ways to test this hypothesis are presented. PMID- 12909694 TI - Optimization of bone growth and remodeling in response to loading in tapered mammalian limbs. AB - How bones respond dynamically to mechanical loading through changes in shape and structure is poorly understood, particularly with respect to variations between bones. Structurally, cortical bones adapt in vivo to their mechanical environments primarily by modulating two processes, modeling and Haversian remodeling. Modeling, defined here as the addition of new bone, may occur in response to mechanical stimuli by altering bone shape or size through growth. Haversian remodeling is thought to be an adaptation to repair microcracks or prevent microcrack propagation. Here, we examine whether cortical bone in sheep limbs modulates periosteal modeling and Haversian remodeling to optimize strength relative to mass in hind-limb midshafts in response to moderate levels of exercise at different growth stages. Histomorphometry was used to compare rates of periosteal growth and Haversian remodeling in exercised and sedentary treatment groups of juvenile, subadult and young adult sheep. In vivo strain data were also collected for the tibia and metatarsal midshafts of juvenile sheep. The results suggest that limb bones initially optimize responses to loading according to the varying power requirements associated with adding mass at different locations. In juveniles, exercise induces higher rates of periosteal modeling in proximal midshafts and higher rates of Haversian remodeling in distal midshafts. Consequently, distal element midshafts experience higher strains and, presumably, have lower safety factors. As animals age, periosteal modeling rates decline and Haversian remodeling rates increase, but moderate levels of mechanical loading stimulate neither process significantly. PMID- 12909695 TI - Kinematics of hovering hummingbird flight along simulated and natural elevational gradients. AB - Hovering flight is one of the most energetically demanding forms of animal locomotion. Despite the cost, hummingbirds regularly hover at high elevations, where flight is doubly challenging because of reduced air density and oxygen availability. We performed three laboratory experiments to examine how air density and oxygen partial pressure influence wingbeat kinematics. In the first study, we experimentally lowered air density but maintained constant oxygen partial pressure. Under these hypodense but normoxic conditions, hummingbirds increased stroke amplitude substantially and increased wingbeat frequency slightly. In the second experiment, we maintained constant air density but decreased oxygen partial pressure. Under these normodense but hypoxic conditions, hummingbirds did not alter stroke amplitude but instead reduced wingbeat frequency until they could no longer generate enough vertical force to offset body weight. In a final combined experiment, we decreased air density but increased oxygen availability, and found that the wingbeat kinematics were unaffected by supplemental oxygen. We also studied hovering and maximally loaded flight performance for 43 hummingbird species distributed along a natural elevational gradient in Peru. During free hovering flight, hummingbirds showed increased stroke amplitude interspecifically at higher elevations, mirroring the intra-individual responses in our first laboratory experiment. During loaded flight, hummingbirds increased both wingbeat frequency and wing stroke amplitude by 19% relative to free-flight values at any given elevation. We conclude that modulation of wing stroke amplitude is a major compensatory mechanism for flight in hypodense or hypobaric environments. By contrast, increases in wingbeat frequency impose substantial metabolic demands, are only elicited transiently and anaerobically, and cannot be used to generate additional sustained lift at high elevations. PMID- 12909696 TI - Metabolite comparisons and the identity of nutrients translocated from symbiotic algae to an animal host. AB - Dinoflagellate algae of the genus Symbiodinium in symbiosis with marine animals release much of their photosynthetic carbon to the animal host. The compounds translocated to the host ('mobile compounds') were investigated by metabolite comparison as follows: a substrate was identified as a candidate mobile compound when comparable profiles of metabolites were generated from host metabolism of this substrate (supplied exogenously) and the endogenous mobile compounds. When the sea anemone Anemonia viridis was incubated with NaH14CO2 under photosynthesizing conditions, most of the radioactivity in the animal tissue was recovered from the low-molecular-mass fraction and distributed in the ratio 1:2:1 between the neutral, acidic and basic sub-fractions. Prominent 14C-labelled compounds included glucose, malate and glucose-6-phosphate. When the symbiosis was incubated with 14C-labelled glucose plus succinate or fumarate (but none of eight other substrate combinations tested), the 14C-labelled metabolites closely matched those obtained with NaH14CO2. These data suggest that glucose and succinate/fumarate (or metabolically allied compounds) may be important photosynthetic compounds transferred from the Symbiodinium cells to the tissues of A. viridis. Metabolite comparisons can be applied to study nutritional interactions in symbioses involving photosynthetic algae and, with appropriate modification, other associations between microorganisms and plants or animals. PMID- 12909697 TI - The effect of colour vision status on the detection and selection of fruits by tamarins (Saguinus spp.). AB - The evolution of trichromatic colour vision by the majority of anthropoid primates has been linked to the efficient detection and selection of food, particularly ripe fruits among leaves in dappled light. Modelling of visual signals has shown that trichromats should be more efficient than dichromats at distinguishing both fruits from leaves and ripe from unripe fruits. This prediction is tested in a controlled captive setting using stimuli recreated from those actually encountered by wild tamarins (Saguinus spp.). Dietary data and reflectance spectra of Abuta fluminum fruits eaten by wild saddleback (Saguinus fuscicollis) and moustached (Saguinus mystax) tamarins and their associated leaves were collected in Peru. A. fluminum leaves, and fruits in three stages of ripeness, were reproduced and presented to captive saddleback and red-bellied tamarins (Saguinus labiatus). Trichromats were quicker to learn the task and were more efficient at selecting ripe fruits than were dichromats. This is the first time that a trichromatic foraging advantage has been demonstrated for monkeys using naturalistic stimuli with the same chromatic properties as those encountered by wild animals. PMID- 12909698 TI - A regulatory role for cortisol in muscle glycogen metabolism in rainbow trout Oncorhynchus mykiss Walbaum. AB - To test the hypothesis that cortisol has a regulatory role in fish muscle glycogenesis post-exercise, rainbow trout were treated 1 h prior to exercise with either saline (control) or metyrapone (2-methyl-1, 2-di-3-pyridyl-1-propanone) to block cortisol synthesis. Following exercise (time 0), half of the metyrapone treated fish received a single injection of cortisol, to mimic the post-exercise rise usually observed. Muscle glycogen and the relative activities of glycogen phosphorylase a (Phos a) and glycogen synthase I (GSase I), regulatory enzymes for glycogen resynthesis, were monitored 4 h post-exercise. Metyrapone treatment succeeded in blocking the post-exercise rise in plasma cortisol (17+/-2 vs 118+/ 13 ng ml(-1) in controls at time 0), and cortisol injection resulted in a larger and more prolonged cortisol increase than in controls (159+/-22 vs 121+/-14 ng ml(-1) in controls at 1 h). Muscle glycogen was completely restored in the metyrapone-treated fish within 2 h after exercise (8.3+/-0.6 vs 8+/-0.7 micromol g(-1) pre-exercise), only partially restored in control fish at 4 h (5.4+/-01.4 vs 8.8+/-1.3 micromol g(-1) pre-exercise), and not at all in cortisol-treated fish (1.0+/-0.5 micromol g(-1) at 4 h). The rapid glycogen resynthesis in the metyrapone-treated fish was associated with a more rapid inactivation of Phos a and stimulation of GSase I compared to controls. In cortisol-treated fish, Phos a activity persisted throughout 4 h post-exercise; there was also a significant stimulation of GSase I activity. As a consequence of dual activation of Phos a and GSase I, glycogen cycling probably occurred, thus preventing net synthesis. This explains why the post-exercise elevation of cortisol inhibits net glycogen synthesis in trout muscle. PMID- 12909699 TI - Load compensation in targeted limb movements of an insect. AB - The task of a multi-jointed limb making an aimed movement towards a target requires that the movement is regulated against external perturbations such as changing load. In particular, loading one part of a limb leads to altered static forces on all proximal segments, and to additional dynamic joint interaction forces when the limb moves. We have addressed the question of load compensation in an insect preparation in which a locust makes aimed scratching movements with a hind leg in response to tactile stimulation of a wing. We show that loading the femur or tibia with the equivalent of 8.5 times the mass of the tibia (corresponding to an increase of up to 11.6 times the rotational moment of inertia at the femur-tibia joint) does not impair the animal's ability to make well-coordinated, aimed movements of that leg towards different targets. The kinematics of the movements are the same, and animals aim the same part of their distal tibia at the target, regardless of loading. The movements are carried out with equal accuracy and at the same initial velocity under all load conditions. Because loading of the leg does not change the behavioural performance, there is no indication of a change in aiming strategy. This implies high leg joint stiffness and/or the existence of high gain proprioceptive control loops. We have previously shown that in the unloaded condition, movements elicited by stimuli to different places on the wing are driven by a single underlying movement pattern that shifts depending on stimulus location along the wing surface. Our present data show that leg proprioceptive inputs are also integrated into the leg motor networks, rendering hind limb targeting robust against large changes in moment of inertia. PMID- 12909700 TI - Temperature and the expression of seven muscle-specific protein genes during embryogenesis in the Atlantic cod Gadus morhua L. AB - Seven cDNA clones coding for different muscle-specific proteins (MSPs) were isolated from the fast muscle tissue of Atlantic cod Gadus morhua L. In situ hybridization using cRNA probes was used to characterize the temporal and spatial patterns of gene expression with respect to somite stage in embryos incubated at 4 degrees C, 7 degrees C and 10 degrees C. MyoD transcripts were first observed in the presomitic mesoderm prior to somite formation, and in the lateral compartment of the forming somites. MyoD expression was not observed in the adaxial cells that give rise to the slow muscle layer, and expression was undetectable by in situ hybridization in the lateral somitic mesoderm after the 35-somite stage, during development of the final approximately 15 somites. RT-PCR analysis, however, confirmed the presence of low levels of the transcript during these later stages. A phylogenetic comparison of the deduced aminoacid sequences of the full-length MyoD cDNA clone and those from other teleosts, and inference from the in situ expression pattern suggested homology with a second paralogue (MyoD2) recently isolated from the gilthead seabream Sparus aurata. Following MyoD expression, alpha-actin was the first structural gene to be switched on at the 16-somite stage, followed by myosin heavy chain, troponin T, troponin I and muscle creatine kinase. The final mRNA in the series to be expressed was troponin C. All genes were switched on prior to myofibril assembly. The troponin C sequence was unusual in that it showed the greatest sequence identity with the rainbow trout Oncorhynchus mykiss cardiac/slow form, but was expressed in the fast myotomal muscle and not in the heart. In addition, the third TnC calcium binding site showed a lower level of sequence conservation than the rest of the sequence. No differences were seen in the timing of appearance or rate of posterior progression (relative to somite stage) of any MSP transcripts between embryos raised at the different temperatures. It was concluded that myofibrillar genes are activated asynchronously in a distinct temporal order prior to myofibrillar assembly and that this process was highly canalized over the temperature range studied. PMID- 12909701 TI - Behavioural investigation of polarisation sensitivity in the Japanese quail (Coturnix coturnix japonica) and the European starling (Sturnus vulgaris). AB - Many animals have sensitivity to the e-vector of linearly polarised light, which may assist in visually mediated behaviours such as navigation, signalling and foraging. However, it is still controversial as to whether birds possess polarisation sensitivity. Several studies have found that altering the polarisation patterns of the broad visual field surrounding birds alters their intended migratory orientation. However, electrophysiological tests have failed to elicit evidence for polarisation sensitivity in birds, and the mechanism by which birds might perceive polarised light is unknown. In this experiment, we trained Japanese quail and European starlings to discriminate stimuli differing in their polarisation pattern. Although both quail and starlings were able to discriminate stimuli in which the stimulus sub-components either differed or had the same radiant intensity (the control task), they were unable to discriminate stimuli in which the e-vector orientations of the stimulus sub-components either differed by 90 degrees or had the same angle of polarisation. The birds' successful performance on the control task, but failure to complete the polarisation task, demonstrated that they had all the necessary cognitive abilities to make the discrimination except sensitivity to angle of polarisation. We conclude that quail and starlings are unable to use polarisation cues in this foraging task. PMID- 12909702 TI - Steroids for free? No metabolic costs of elevated maternal androgen levels in the black-headed gull. AB - Within- and between-clutch variation in yolk titres of hormones of maternal origin has been found in many avian species. So far, experiments have revealed mainly beneficial effects of maternal androgens. This would also apply to black headed gulls (Larus ridibundus). Previous experiments have shown that chicks benefit from these higher levels since their competitive abilities are improved and growth and survival probabilities thus enhanced. However, not all females show the same increase in yolk hormones from first to last egg or invest equally high amounts of androgens in their clutches. Possibly, there is a trade-off between the beneficial effects of high androgen levels and potential costs, such as increased metabolic rates. We studied possible metabolic costs of experimentally elevated yolk androgen levels for chicks of several age classes, starting three days prior to hatching until fledging at an age of approximately 30 days. Daily energy expenditure in the field, measured using the doubly labelled water technique, did not differ between treatments or between sexes. Oxygen consumption measured in birds at rest in the lab (RMR) did not vary between chicks hatched from androgen-injected (T) or oil-injected (Oil) control eggs at any age in thermo-neutral or below thermo-neutral conditions. Males showed a lower RMR than females towards the fledging age. We conclude that it is unlikely that the costs of high maternal androgen levels can be found in higher energy expenditure in the chick. PMID- 12909703 TI - Hepoxilins and trioxilins in barnacles: an analysis of their potential roles in egg hatching and larval settlement. AB - The barnacle life cycle has two key stages at which eicosanoids are believed to be involved in cellular communication pathways, namely the hatching of nauplii and the settlement of cypris larvae. Barnacle egg-hatching activity has previously been reported to reside in a variety of eicosanoids, including 8 hydroxyeicosapentaenoic acid and a number of tri-hydroxylated polyunsaturated fatty acid derivatives, the trioxilins. The production of the eicosapentaenoic acid metabolite trioxilin A4 (8,11,12-trihydroxy-5,9,14,17-eicosatetraenoic acid) by the barnacles Balanus amphitrite and Elminius modestus was confirmed using a combination of high-performance liquid chromatography and gas chromatography, both linked to mass spectrometry. In addition, both species also generated trioxilin A3 (8,11,12-trihydroxy-5,9,14-eicosatrienoic acid; an arachidonic acid derived product), 8,11,12-trihydroxy-9,14,17-eicosatrienoic acid (a omega3 analogue of trioxilin A3; derived from omega3 arachidonic acid) and 10,13,14 trihydroxy-4,7,11,16,19-docosapentaenoic acid (a docosahexaenoic acid-derived product). In contrast to earlier reports, trioxilin A3 had no E. modestus egg hatching activity at any of the concentrations tested (10(-9)-10(-6) mol l(-1)). The unstable epoxide precursor hepoxilin A3, however, caused significant levels of hatching at 10(-6) mol l(-1). Furthermore, the stable hepoxilin B3 analogue PBT-3 stimulated hatching at 10(-7) mol l(-1). Neither trioxilin A3, hepoxilin A3 or PBT-3 at 0.25-30 micromol l(-1) served as settlement cues for B. amphitrite cypris larvae. PMID- 12909705 TI - The effect of temperature on swimming performance and oxygen consumption in adult sockeye (Oncorhynchus nerka) and coho (O. kisutch) salmon stocks. AB - Our knowledge of the swimming capabilities and metabolic rates of adult salmon, and particularly the influence of temperature on them, is extremely limited, and yet this information is critical to understanding the remarkable upstream migrations that these fish can make. To remedy this situation, we examined the effects of temperature on swimming performance and metabolic rates of 107 adult fish taken from three stocks of sockeye salmon Oncorhynchus nerka and one stock of coho salmon O. kisutch at various field and laboratory locations, using large, portable, swim tunnels. The salmon stocks were selected because of differences in their ambient water temperature (ranging from 5 degrees C to 20 degrees C) and the total distance of their in-river migrations (ranging from approximately 100 km for coastal stocks to approximately 1100 km for interior stocks). As anticipated, differences in routine metabolic rate observed among salmon stocks were largely explained by an exponential dependence on ambient water temperature. However, the relationship between water temperature and maximum oxygen consumption (MO2max), i.e. the MO2 measured at the critical swimming speed (Ucrit), revealed temperature optima for MO2max that were stock-specific. These temperature optima were very similar to the average ambient water temperatures for the natal stream of a given stock. Furthermore, at a comparable water temperature, the salmon stocks that experienced a long and energetically costly in-river migration were characterized by a higher MO2max, a higher scope for activity, a higher Ucrit and, in some cases, a higher cost of transport, relative to the coastal salmon stocks that experience a short in-river migration. We conclude that high-caliber respirometry can be performed in a field setting and that stock-specific differences in swimming performance of adult salmon may be important for understanding upstream migration energetics and abilities. PMID- 12909704 TI - Biochemical support for the V-ATPase rotary mechanism: antibody against HA-tagged Vma7p or Vma16p but not Vma10p inhibits activity. AB - V-ATPase null mutants in yeast have a distinct, conditionally lethal phenotype that can be obtained through disruption of any one of its subunits. This enables supplementation of this mutant with the relevant subunit tagged with an epitope against which an antibody is available. In this system, the effect of antibody on the activity of the enzyme can be analyzed. Towards this end we used HA to tag subunits Vma7p, Vma10p and Vma16p, which are assumed to represent, respectively, the shaft, stator and turbine of the enzyme, and used them to supplement the corresponding yeast V-ATPase null mutants. The anti-HA epitope antibody inhibited both the ATP-dependent proton uptake and the ATPase activities of the Vma16p-HA and Vma7p-HA containing complexes, in intact vacuoles and in the detergent solubilized enzyme. Neither of these activities was inhibited by the antibody in Vma10p-HA containing enzyme. These results support the function of Vma10p as part of the stator, while the other tagged subunits are part of the rotor apparatus. The HA-tag was attached to the N terminus of Vma16p; thus the antibody inhibition points to its accessibility outside the vacuolar membrane. This assumption is supported by the supplementation of the yeast mutant by the homologues of Vma16p isolated from Arabidopsis thaliana and lemon fruit c-DNA. Contrary to yeast, which has five predicted helices, the plant subunit Vma16p has only four. Our results confirm a recent report that only four of the yeast Vma16p complexes are actually transmembrane helices. PMID- 12909706 TI - Excess post-exercise oxygen consumption in adult sockeye (Oncorhynchus nerka) and coho (O. kisutch) salmon following critical speed swimming. AB - The present study measured the excess post-exercise oxygen cost (EPOC) following tests at critical swimming speed (Ucrit) in three stocks of adult, wild, Pacific salmon (Oncorhynchus sp.) and used EPOC to estimate the time required to return to their routine level of oxygen consumption (recovery time) and the total oxygen cost of swimming to Ucrit. Following exhaustion at Ucrit, recovery time was 42-78 min, depending upon the fish stock. The recovery times are several-fold shorter than previously reported for juvenile, hatchery-raised salmonids. EPOC varied fivefold among the fish stocks, being greatest for Gates Creek sockeye salmon (O. nerka), which was the salmon stock that had the longest in-river migration, experienced the warmest temperature and achieved the highest maximum oxygen consumption compared with the other salmon stocks that were studied. EPOC was related to Ucrit, which in turn was directly influenced by ambient test temperature. The non-aerobic cost of swimming to Ucrit was estimated to add an additional 21.4-50.5% to the oxygen consumption measured at Ucrit. While these non-aerobic contributions to swimming did not affect the minimum cost of transport, they were up to three times higher than the value used previously for an energetic model of salmon migration in the Fraser River, BC, Canada. As such, the underestimate of non-aerobic swimming costs may require a reevaluation of the importance of how in-river barriers like rapids and bypass facilities at dams, and year-to-year changes in river flows and temperatures, affect energy use and hence migration success. PMID- 12909707 TI - Retinoid X receptor and retinoic acid response in the marine sponge Suberites domuncula. AB - To date no nuclear receptors have been identified or cloned from the phylogenetically oldest metazoan phylum, the Porifera (sponges). We show that retinoic acid causes tissue regression in intact individuals of the demosponge Suberites domuncula and in primmorphs, special three-dimensional cell aggregates. Primmorphs were cultivated on a galectin/poly-L-lysine matrix in order to induce canal formation. In the presence of 1 or 50 micromol l(-1) retinoic acid these canals undergo regression, a process that is reversible. We also cloned the cDNA from S. domuncula encoding the retinoid X receptor (RXR), which displays the two motifs of nuclear hormone receptors, the ligand-binding and the DNA-binding domains, and performed phylogenetic analyses of this receptor. RXR expression undergoes strong upregulation in response to treatment with retinoic acid, whereas the expression of the sponge caspase is not increased. The gene encoding the LIM homeodomain protein was found to be strongly upregulated in response to retinoic acid treatment. These data indicate that the RXR and its ligand retinoic acid play a role in the control of morphogenetic events in sponges. PMID- 12909708 TI - Comparison of renal and salt gland function in three species of wild ducks. AB - Three processes central to osmoregulation of marine birds were compared in three species of ducks that differ in habitat affinity, diet and saline tolerance. These processes are filtration of Na+ and water from the plasma by the kidneys, their reabsorption along the renal tubules, and secretion by the salt glands. Barrow's goldeneyes Bucephala islandica, the most marine species, have the highest rates for all three processes and only this species can secrete all the infused salt via the salt glands. Rates of all three processes are lower in mallards Anas platyrhynchos, the most freshwater species. Following saline acclimation, mallards could excrete all the infused Na+ by a combined Na+ excretion of the kidneys and salt glands. Canvasbacks Aythya valisineria, despite being more saline tolerant than mallards, are unable to excrete all the infused Na+. They produce a large volume of urine (like mallards) that has a low [Na+] (like goldeneyes). Salt gland secretion Na+ concentration did not differ among the three species, but only goldeneyes secrete at a rate sufficient to eliminate all infused Na+ via the salt glands. Differences in saline tolerance of these ducks species cannot be fully explained by differences in their filtration, reabsorption and secretion of Na+ and water, suggesting that the intestinal tract plays an important role. PMID- 12909709 TI - Regulation of a renal urea transporter with reduced salinity in a marine elasmobranch, Raja erinacea. AB - Marine elasmobranchs retain urea and other osmolytes, e.g. trimethylamine oxide (TMAO), to counterbalance the osmotic pressure of seawater. We investigated whether a renal urea transporter(s) would be regulated in response to dilution of the external environment. A 779 bp cDNA for a putative skate kidney urea transporter (SkUT) was cloned, sequenced and found to display relatively high identity with facilitated urea transporters from other vertebrates. Northern analysis using SkUT as a probe revealed three signals in the kidney at 3.1, 2.8 and 1.6 kb. Upon exposure to 50% seawater, the levels of all three SkUT transcripts were significantly diminished in the kidney (by 1.8- to 3.5-fold). In response to environmental dilution, renal tissue osmolality and urea concentration decreased, whereas water content increased. There were no significant differences in osmolyte and mRNA levels between the dorsal-lateral bundle and ventral sections of the kidney. Taken together, these findings provide evidence that the downregulation of SkUT may play a key role in lowering tissue urea levels in response to external osmolality. PMID- 12909710 TI - Urea transport in kidney brush-border membrane vesicles from an elasmobranch, Raja erinacea. AB - Marine elasmobranch fishes maintain high urea concentrations and therefore must minimize urea loss to the environment in order to reduce the energetic costs of urea production. Previous studies have identified a facilitated urea transporter in the kidney of the dogfish. We examined mechanisms of urea transport in the kidney of the little skate Raja erinacea using an isolated brush-border membrane vesicle preparation. Urea uptake by brush-border membrane vesicles is by a phloretin-sensitive, non-saturable uniporter in the dorsal section and a phloretin-sensitive, sodium-linked urea transporter (Km = 0.70 mmol l(-1), Vmax = 1.18 micromol h(-1) mg(-1) protein) in the ventral section of the kidney. This provides evidence for two separate urea transporters in the dorsal versus ventral sections of the kidney. We propose that these two mechanisms of urea transport are critical for renal urea reabsorption in the little skate. PMID- 12909711 TI - Activation of Notch signaling pathway precedes heart regeneration in zebrafish. AB - Several vertebrates display the ability to regenerate parts of their body after amputation. During this process, differentiated cells reenter the cell cycle and proliferate to generate a mass of undifferentiated cells. Repatterning mechanisms act on these cells to eventually shape a regenerated tissue or organ that replaces the amputated one. Experiments with regenerating limbs/fins in newts and zebrafish have shown that members of the Msx family of homeodomain-containing transcription factors play key roles during blastema formation and patterning. Here we show that adult zebrafish have a remarkable capacity to regenerate the heart in a process that involves up-regulation of msxB and msxC genes. We present evidence indicating that heart regeneration involves the execution of a specific genetic program, rather than redeployment of a cardiac development program. Preceding Msx activation, there is a marked increase in the expression of notch1b and deltaC, which we show are also up-regulated during fin regeneration. These data suggest a role for the Notch pathway in the activation of the regenerative response. Taken together, our results underscore the use of zebrafish as a model for investigating the process of regeneration in particular and the biology of stem cells in general. Advances in these fields will undoubtedly aid in the implementation of strategies for regenerative medicine. PMID- 12909712 TI - High concentrations of long interspersed nuclear element sequence distinguish monoallelically expressed genes. AB - Genes subject to monoallelic expression are expressed from only one of the two alleles either selected at random (random monoallelic genes) or in a parent-of origin specific manner (imprinted genes). Because high densities of long interspersed nuclear element (LINE)-1 transposon sequence have been implicated in X-inactivation, we asked whether monoallelically expressed autosomal genes are also flanked by high densities of LINE-1 sequence. A statistical analysis of repeat content in the regions surrounding monoallelically and biallelically expressed genes revealed that random monoallelic genes were flanked by significantly higher densities of LINE-1 sequence, evolutionarily more recent and less truncated LINE-1 elements, fewer CpG islands, and fewer base-pairs of short interspersed nuclear elements (SINEs) sequence than biallelically expressed genes. Random monoallelic and imprinted genes were pooled and subjected to a clustering analysis algorithm, which found two clusters on the basis of aforementioned sequence characteristics. Interestingly, these clusters did not follow the random monoallelic vs. imprinted classifications. We infer that chromosomal sequence context plays a role in monoallelic gene expression and may involve the recognition of long repeats or other features. The sequence characteristics that distinguished the high-LINE-1 category were used to identify more than 1,000 additional genes from the human and mouse genomes as candidate genes for monoallelic expression. PMID- 12909713 TI - In vivo regeneration of murine prostate from dissociated cell populations of postnatal epithelia and urogenital sinus mesenchyme. AB - The existence of a postnatal prostate stem cell is supported by several types of evidence. Withdrawal of androgen leads to involution of the gland, but readdition can rapidly stimulate regeneration. Tissue fragments derived from mouse or rat prostatic epithelia from midgestation embryos or adult mice, when combined with tissue fragments from urogenital sinus mesenchyme and grafted under the kidney capsule, can regenerate prostatic structures. Indirect evidence supports that the stem cell population is contained within the basal layer. Purified prostatic stem cell preparations would be useful to define the physical and functional properties required for regeneration and to compare with cells that accumulate during abnormal growth states, like prostate cancer. We have developed a regeneration system using dissociated cell populations of postnatal prostate epithelia and embryonic urogenital sinus mesenchyme. Efficient in vivo regeneration of prostatic structures in the subcapsular space of the kidney was observed within 4-8 wk with as few as 103 epithelial cells from prostates derived from donors 10 d to 6 wk of age. The regenerated structures show a branching tubular epithelial morphology, with expression of a panel of markers consistent with prostate development. Donor epithelial populations can be readily infected with GFP expressing lentiviral vectors to provide integration markers and easy visualization. The cell preparations of urogenital sinus mesenchyme can be expanded in short-term in vitro culture while their inductive capabilities are retained. Further definition of the subpopulation of prostate epithelial cells containing the regeneration activity should be possible with such technologies. PMID- 12909714 TI - Long-distance root-to-shoot transport of phytochelatins and cadmium in Arabidopsis. AB - Phytochelatin synthases (PCS) mediate cellular heavy-metal resistance in plants, fungi, and worms. However, phytochelatins (PCs) are generally considered to function as intracellular heavy-metal detoxification mechanisms, and whether long distance transport of PCs occurs during heavy-metal detoxification remains unknown. Here, wheat TaPCS1 cDNA expression was either targeted to Arabidopsis roots with the Arabidopsis alcohol dehydrogenase (Adh) promoter (Adh::TaPCS1/cad1 3) or ectopically expressed with the cauliflower mosaic virus 35S promoter (35S::TaPCS1/cad1-3) in the PC-deficient mutant cad1-3. Adh::TaPCS1/cad1-3 and 35S::TaPCS1/cad1-3 complemented the cadmium, mercury, and arsenic sensitivities of the cad1-3 mutant. Northern blot, RT-PCR, and Western blot analyses showed Adh promoter-driven TaPCS1 expression only in roots and thus demonstrated lack of long-distance TaPCS1 mRNA and protein transport in plants. Fluorescence HPLC analyses showed that under Cd2+ stress, no PCs were detectable in cad1-3. However, in Adh::TaPCS1/cad1-3 plants, PCs were detected in roots and in rosette leaves and stems. Inductively coupled plasma atomic emission spectrometer analyses showed that either root-specific or ectopic expression of TaPCS1 significantly enhanced long-distance Cd2+ transport into stems and rosette leaves. Unexpectedly, transgenic expression of TaPCS1 reduced Cd2+ accumulation in roots compared with cad1-3. The reduced Cd2+ accumulation in roots and enhanced root-to-shoot Cd2+ transport in transgenic plants were abrogated by l buthionine sulfoximine. The presented findings show that (i) transgenic expression of TaPCS1 suppresses the heavy-metal sensitivity of cad1-3, (ii) PCs can be transported from roots to shoots, and (iii) transgenic expression of the TaPCS1 gene increases long-distance root-to-shoot Cd2+ transport and reduces Cd2+ accumulation in roots. PMID- 12909715 TI - Glutathione synthetase homologs encode alpha-L-glutamate ligases for methanogenic coenzyme F420 and tetrahydrosarcinapterin biosyntheses. AB - Proteins in the ATP-grasp superfamily of amide bond-forming ligases have evolved to function in a number of unrelated biosynthetic pathways. Previously identified homologs encoding glutathione synthetase, d-alanine:d-alanine ligase and the bacterial ribosomal protein S6:glutamate ligase have been vertically inherited within certain organismal lineages. Although members of this specificity-diverse superfamily share a common reaction mechanism, the nonoverlapping set of amino acid and peptide substrates recognized by each family provided few clues as to their evolutionary history. Two members of this family have been identified in the hyperthermophilic marine archaeon Methanococcus jannaschii and shown to catalyze the final reactions in two coenzyme biosynthetic pathways. The MJ0620 (mptN) locus encodes a tetrahydromethanopterin:alpha-l-glutamate ligase that forms tetrahydrosarcinapterin, a single carbon-carrying coenzyme. The MJ1001 (cofF) locus encodes a gamma-F420-2:alpha-l-glutamate ligase, which caps the gamma-glutamyl tail of the hydride carrier coenzyme F420. These two genes share a common ancestor with the ribosomal protein S6:glutamate ligase and a putative alpha-aminoadipate ligase, defining the first group of ATP-grasp enzymes with a shared amino acid substrate specificity. As in glutathione biosynthesis, two unrelated amino acid ligases catalyze sequential reactions in coenzyme F420 polyglutamate formation: a gamma-glutamyl ligase adds 1-3 l-glutamate residues and the ATP-grasp-type ligase described here caps the chain with a single alpha linked l-glutamate residue. The analogous pathways for glutathione, F420, folate, and murein peptide biosyntheses illustrate convergent evolution of nonribosomal peptide biosynthesis through the recruitment of single-step amino acid ligases. PMID- 12909716 TI - Atypical expansion in mice of the sensory neuron-specific Mrg G protein-coupled receptor family. AB - The Mas-related genes (Mrgs) comprise a family of >50 G protein-coupled receptors (GPCRs), many of which are expressed in specific subsets of nociceptive sensory neurons in mice. In contrast, humans contain a related but nonorthologous family of genes, called MrgXs or sensory neuron-specific receptors, of which many fewer appear to be expressed in sensory neurons. To determine whether the diversity of murine Mrgs is generic to rodents or is an atypical feature of mice, we characterized MrgA, MrgB, MrgC, and MrgD subfamilies in rat and gerbil. Surprisingly, although mice have approximately 22 MrgA and approximately 14 MrgC genes, rats and gerbils have just a single MrgA and MrgC gene. This murine specific expansion likely reflects recent retrotransposon-mediated unequal crossover events. The expression of Mrgs in rat sensory ganglia suggests that the extensive cellular diversity in mice can be simplified to a core subset of approximately four different genes (MrgA, MrgB, MrgC, and MrgD), defining a similar number of neuronal subpopulations. Our results suggest more generally that mouse-human genomic comparisons may sometimes reveal differences atypical of rodents. PMID- 12909717 TI - Defective double-strand DNA break repair and chromosomal translocations by MYC overexpression. AB - DNA repair mechanisms are essential for the maintenance of genomic integrity. Disruption of gene products responsible for DNA repair can result in chromosomal damage. Improperly repaired chromosomal damage can result in the loss of chromosomes or the generation of chromosomal deletions or translocations, which can lead to tumorigenesis. The MYC protooncogene is a transcription factor whose overexpression is frequently associated with human neoplasia. MYC has not been previously implicated in a role in DNA repair. Here we report that the overexpression of MYC disrupts the repair of double-strand DNA breaks, resulting in a several-magnitude increase in chromosomal breaks and translocations. We found that MYC inhibited the repair of gamma irradiation DNA breaks in normal human cells and blocked the repair of a single double-strand break engineered to occur in an immortal cell line. By spectral karyotypic analysis, we found that MYC even within one cell division cycle resulted in a several-magnitude increase in the frequency of chromosomal breaks and translocations in normal human cells. Hence, MYC overexpression may be a previously undescribed example of a dominant mutator that may fuel tumorigenesis by inducing chromosomal damage. PMID- 12909718 TI - Nuclear import of hepatitis B virus capsids and release of the viral genome. AB - While studying the import of the hepatitis B virus genome into the nucleus of permeabilized tissue culture cells, we found that viral capsids were imported in intact form through the nuclear pore into the nuclear basket. Import depended on phosphorylation of the capsid protein and was mediated by the cellular transport receptors importin alpha and beta. Virus-derived capsids that contained the mature viral genome were able to release the viral DNA and capsid protein into the nucleoplasm. The uncoating reaction was independent of Ran, a GTP-binding enzyme responsible for dissociating other imported cargo from the inner face of the nuclear pore. Immature capsids that did not contain the mature viral genome reached the basket but did not release capsid proteins nor immature genomes into the nucleoplasm. The different fate of mature and immature capsids after passing the nuclear pore indicates that the outcome of a nuclear import event may be regulated within the nuclear basket. PMID- 12909719 TI - Separating pitch chroma and pitch height in the human brain. AB - Musicians recognize pitch as having two dimensions. On the keyboard, these are illustrated by the octave and the cycle of notes within the octave. In perception, these dimensions are referred to as pitch height and pitch chroma, respectively. Pitch chroma provides a basis for presenting acoustic patterns (melodies) that do not depend on the particular sound source. In contrast, pitch height provides a basis for segregation of notes into streams to separate sound sources. This paper reports a functional magnetic resonance experiment designed to search for distinct mappings of these two types of pitch change in the human brain. The results show that chroma change is specifically represented anterior to primary auditory cortex, whereas height change is specifically represented posterior to primary auditory cortex. We propose that tracking of acoustic information streams occurs in anterior auditory areas, whereas the segregation of sound objects (a crucial aspect of auditory scene analysis) depends on posterior areas. PMID- 12909721 TI - ELG1, a yeast gene required for genome stability, forms a complex related to replication factor C. AB - Many overlapping surveillance and repair mechanisms operate in eukaryotic cells to ensure the stability of the genome. We have screened to isolate yeast mutants exhibiting increased levels of recombination between repeated sequences. Here we characterize one of these mutants, elg1. Strains lacking Elg1p exhibit elevated levels of recombination between homologous and nonhomologous chromosomes, as well as between sister chromatids and direct repeats. These strains also exhibit increased levels of chromosome loss. The Elg1 protein shares sequence homology with the large subunit of the clamp loader replication factor C (RFC) and with the product of two additional genes involved in checkpoint functions and genome maintenance: RAD24 and CTF18. Elg1p forms a complex with the Rfc2-5 subunits of RFC that is distinct from the previously described RFC-like complexes containing Rad24 and Ctf18. Genetic data indicate that the Elg1, Ctf18, and Rad24 RFC-like complexes work in three separate pathways important for maintaining the integrity of the genome and for coping with various genomic stresses. PMID- 12909720 TI - Functional mutants of the sequence-specific transcription factor p53 and implications for master genes of diversity. AB - There are many sources of genetic diversity, ranging from programmed mutagenesis in antibody genes to random mutagenesis during species evolution or development of cancer. We propose that mutations in DNA sequence-specific transcription factors that target response elements (REs) in many genes can also provide for rapid and broad phenotypic diversity, if the mutations lead to altered binding affinities at individual REs. To test this concept, we examined the in vivo transactivation capacity of wild-type human and murine p53 and 25 partial function mutants. The p53s were expressed in yeast from a rheostatable promoter, and the transactivation capacities toward >15 promoter REs upstream of a reporter gene were measured. Surprisingly, there was wide variation in transactivation by the mutant p53s toward the various REs. This is the first study to address directly the impact of mutations in a sequence-specific transcription factor on transactivation from a wide array of REs. We propose a master gene hypothesis for phenotypic diversity where the master gene is a single transcriptional activator (or repressor) that regulates many genes through different REs. Mutations of the master gene can lead to a variety of simultaneous changes in both the selection of targets and the extent of transcriptional modulation at the individual targets, resulting in a vast number of potential phenotypes that can be created with minimal mutational changes without altering existing protein-protein interactions. PMID- 12909722 TI - Polar auxin transport in the wood-forming tissues of hybrid aspen is under simultaneous control of developmental and environmental signals. AB - Recent research has highlighted the importance of auxin concentration gradients during plant development. Establishment of these gradients is believed to involve polar auxin transport through specialized carrier proteins. We have used an experimental system, the wood-forming tissue of hybrid aspen, which allows tissue specific expression analysis of auxin carrier genes and quantification of endogenous concentrations of the hormone. As part of this study, we isolated the putative polar auxin transport genes, PttLAX1-PttLAX3 and PttPIN1-PttPIN3, belonging to the AUX1-like family of influx and PIN1-like efflux carriers, respectively. Analysis of PttLAX and PttPIN expression suggests that specific positions in a concentration gradient of the hormone are associated with different stages of vascular cambium development and expression of specific members of the auxin transport gene families. We were also able demonstrate positive feedback of auxin on polar auxin transport genes. Entry into dormancy at the end of a growing season leads to a loss of auxin transport capacity, paralleled by reduced expression of PttLAX and PttPIN genes. Furthermore, data from field experiments show that production of the molecular components of the auxin transport machinery is governed by environmental controls. Our findings collectively demonstrate that trees have developed mechanisms to modulate auxin transport in the vascular meristem in response to developmental and environmental cues. PMID- 12909724 TI - Optimal design of a bistable switch. AB - Determining optimally designed structures is important for diverse fields of science and engineering. Here we describe a procedure for calculating the optimal design of a switch and apply the method to a bistable microelectromechanical system relay switch. The approach focuses on characterizing the unstable transition state connecting the two stable equilibria to control the force displacements. Small modifications in component shape lead to a substantial improvement in device operation. Fabrication of the optimized devices confirms the predictions. PMID- 12909723 TI - The 15-lipoxygenase-1 product 13-S-hydroxyoctadecadienoic acid down-regulates PPAR-delta to induce apoptosis in colorectal cancer cells. AB - Diminished apoptosis, a critical event in tumorigenesis, is linked to down regulated 15-lipoxygenase-1 (15-LOX-1) expression in colorectal cancer cells. 13 S-hydroxyoctadecadienoic acid (13-S-HODE), which is the primary product of 15-LOX 1 metabolism of linoleic acid, restores apoptosis. Nonsteroidal antiinflammatory drugs (NSAIDs) transcriptionally up-regulate 15-LOX-1 expression to induce apoptosis. Peroxisome proliferator-activated receptors (PPARs) are nuclear receptors for linoleic and arachidonic acid metabolites. PPAR-delta promotes colonic tumorigenesis. NSAIDs suppress PPAR-delta activity in colon cancer cells. The mechanistic relationship between 15-LOX-1 and PPAR-delta was previously unknown. Our current study shows that (i) 13-S-HODE binds to PPAR-delta, decreases PPAR-delta activation, and down-regulates PPAR-delta expression in colorectal cancer cells; (ii) the induction of 15-LOX-1 expression is a critical step in NSAID down-regulation of PPAR-delta and the resultant induction of apoptosis; and (iii) PPAR-delta is an important signaling receptor for 13-S-HODE induced apoptosis. The in vivo relevance of these mechanistic findings was demonstrated in our tumorigenesis studies in nude mouse xenograft models. Our findings indicate that the down-regulation of PPAR-delta by 15-LOX-1 through 13-S HODE is an apoptotic signaling pathway that is activated by NSAIDs. PMID- 12909725 TI - Targeted gene evolution in Escherichia coli using a highly error-prone DNA polymerase I. AB - We present a system for random mutagenesis in Escherichia coli for the evolution of targeted genes. To increase error rates of DNA polymerase I (Pol I) replication, we introduced point mutations in three structural domains that govern Pol I fidelity. Expression of error-prone Pol I in vivo results in strong mutagenesis of a target sequence encoded in a Pol I-dependent plasmid (8.1 x 10-4 mutations per bp, an 80,000-fold increase), with a preference for plasmid relative to chromosome sequence. Mutagenesis is maximal in cultures maintained at stationary phase. Mutations are evenly distributed and show a variety of base pair substitutions, predominantly transitions. Mutagenesis extends at least 3 kb beyond the 400-500 nt reportedly synthesized by Pol I. We demonstrate that our error-prone Pol I can be used to generate enzymes with distinct properties by generating TEM-1 beta-lactamase mutants able to hydrolyze a third-generation lactam antibiotic, aztreonam. Three different mutations contribute to aztreonam resistance. Two are found in the extended-spectrum beta-lactamases most frequently identified in clinical isolates, and the third (G276R) has not been previously described. Our system of targeted mutagenesis in E. coli should have an impact on enzyme-based applications in areas such as synthetic chemistry, gene therapy, and molecular biology. Given the structural conservation between polymerases, this work should also provide a reference for altering the fidelity of other polymerases. PMID- 12909726 TI - Long-range enhancer-promoter interactions in the Scr-Antp interval of the Drosophila Antennapedia complex. AB - Long-range enhancer-promoter interactions are commonly seen in complex genetic loci such as Hox genes and globin genes. In the case of the Drosophila Antennapedia complex, the T1 enhancer bypasses the neighboring ftz gene and interacts with the distant Scr promoter to activate expression in posterior head segments. Previous studies identified a 450-bp promoter-proximal sequence, the tethering element, which is essential for T1-Scr interactions. To obtain a more comprehensive view of how individual enhancers selectively interact with appropriate target genes, we used bioinformatic methods to identify new cis regulatory DNAs in the approximately 50-kb Scr-Antp interval. Three previously uncharacterized regulatory elements were identified: a distal T1 tethering sequence mapping >40 kb from the proximal tethering sequence, a repressor element that excludes activation of Scr by inappropriate enhancers, and a new ftz enhancer that directs expression within the limits of stripes 1 and 5. Many of the regulatory DNAs in the Scr-Antp interval are transcribed, including the proximal and distal tethering elements. We suggest that homotypic interactions between the tethering elements stabilize long-range T1-Scr interactions during development. PMID- 12909727 TI - Horseradish peroxidase immobilized through its carboxylic groups onto a polyacrylonitrile membrane: comparison of enzyme performances with inorganic beaded supports. AB - A hydrophilic polyacrylonitrile (PAN) flat sheet membrane was aminated (8.5 micromol of NH2/mg of dry support) for covalent binding of horseradish peroxidase (HRP), mediated by the soluble carbodiimide l-ethyl-3-(3- dimethylaminopropyl)carbodiimide (EDC). Silica microbeads derivatized by silanization, to yield an aminated support, and commercial aminated glass microbeads were also coupled to HRP with EDC or activated with glutaraldehyde. The immobilized enzyme activities were determined in a batch enzyme reactor with an external loop, the highest specific immobilized HRP activity being obtained on the glass support (55.8 U/mg of protein). Continuous operational stability studies showed that hydrophilic PAN membrane led to the highest retention of HRP activity after an overall period of 35 h, with a normalized productivity of 59.5 micromol of H2O2 reduced/(h x Uimmob HRP). PMID- 12909728 TI - L-asparaginase release from Escherichia coli cells with aqueous two-phase micellar systems. AB - A method was proposed to release and separate L-asparaginase (EC 3.5.1.1) from Escherichia coli ATCC 11303 cells with aqueous two-phase micellar systems. The systems were composed of K2HPO4 and Triton X-100. The method combines enzyme release with enzyme purification. The influence of Triton X-100 concentration, K2HPO4 concentration, and pH on the release and partition of L-asparaginase was investigated. Experimental results showed that E. coli cells treated with 9.4% (w/v) K2HPO4 and 15% (w/v) Triton X-100 at 25 degrees C for 15-20 h released nearly 80% of the enzyme. Most of the released enzyme was partitioned to the bottom phase (phosphate-rich phase). The effects of Triton X-100 concentration, K2HPO4 concentration, and pH on cloud point were also studied. Electron micrography indicated that the chemical treatment altered the inner structure of E. coli cells significantly. PMID- 12909730 TI - Effect of tributylphenyltetraethoxylate on enzyme production of Pleurotus ostreatus. AB - The effect of the addition of the nonionic surfactant tributylphenyltetraethoxylate to culture media on pH and extracellular protein content, and on production of beta-glucosidase, xylanase, laccase, and manganese dependent and -independent peroxidases by the edible fungus Pleurotus ostreatus was determined. The relationship between fermentation parameters and concentration of surfactant was assessed by multiple linear regression analysis, and the similarities and differences among the fermentation parameters were elucidated by principal component analysis. Calculations proved that except for xylanase all other cultivation parameters were significantly influenced by the surfactant, with the effect higher at higher surfactant concentrations. Surfactant increased the production of beta-glucosidase and inhibited laccase and manganese-dependent and -independent peroxidase activities. PMID- 12909729 TI - Solid-phase reducing agents as alternative for reducing disulfide bonds in proteins. AB - Disulfide reduction of Kluyveromyces lactis and Aspergillus oryzae beta galactosidases and beta-lactoglobulin was assessed. Reduction was performed using one of two thiol-containing agents: dithiothreitol (DTT) or thiopropyl-agarose with a high degree of substitution (1000 micromol of SH groups/g of dried gel). Both reductants allowed an increase of three- (for K. lactis beta-galactosidase) and fourfold (for A. oryzae beta-galactosidase) in the initial content of SH groups in the lactases. Nearly sevenfold fewer micromoles of SH groups per milligram of protein were needed to perform the reduction of K. lactis beta galactosidase with thiopropyl-agarose than for the same reduction with DTT. However, for A. oryzae beta-galactosidase, nearly twice as many micromoles of SH groups per milligram of protein were needed with thiopropylagarose than with DTT. Disulfide bonds in beta-lactoglobulin were not accessible to thiopropyl-agarose, since this reduction was only possible in the presence of 6 M urea. These results proved that highly substituted thiopropyl-agarose is as good a reducing agent as DTT, for the reduction of disulfide bonds in proteins. Moreover, excess reducing agent was very simply separated from the reduced protein by filtration, making it easier to control the reaction and providing reduced protein solutions free of reductant. All these advantages substantially cut down the time required and therefore the cost of the overall process. PMID- 12909732 TI - Calcium alginate gel as encapsulation matrix for coimmobilized enzyme systems. AB - Encapsulation within calcium alginate gel capsules was used to produce a coimmobilized enzyme system. Glucose oxidase (GOD) and catalase (CAT) were chosen as model enzymes. The same values of Vmax and Km app for the GOD encapsulated system and for the GOD-CAT coencapsulated system were calculated. When gel beads and capsules were compared, the same catalyst deactivation sequence for the two enzymes was observed. However, when capsules were employed as immobilization support, GOD efficiencies were higher than for the gel beads. These results were explained in terms of the structure of the capsules. PMID- 12909731 TI - Effect of enzymatic interesterification on melting point of palm olein. AB - Immobilized PS-C 'Amano' II lipase was used to catalyze the interesterification of palm olein (POo) with 30, 50, and 70% stearic acid in n-hexane at 60 degrees C. The catalytic performance of the immobilized lipase was evaluated by determining the composition change of fatty acyl groups and triacylglycerol (TAG) by gas liquid chromatography and high-performance liquid chromatography, respectively. The interesterification process resulted in the formation of new TAGs, mainly tripalmitin and dipalmitostearin, both of which were absent in the original oil. These changes in TAG composition resulted in an increase in slip melting point, from the original 25.5 degrees C to 36.3, 37.0, and 40.0 degrees C in the modified POo with 30, 50, and 70% stearic acid, respectively. All the reactions attained steady state in about 6 h. This type of work will find great applications in food industries, such as confectionery. PMID- 12909733 TI - The challenge of pancreatic cancer. PMID- 12909734 TI - Molecular epidemiology of pancreatic cancer. AB - Pancreatic cancer is the fourth leading cause of cancer-related death in the United States. Currently there is no early diagnostic test and no effective treatment options for this deadly disease. Prevention of pancreatic cancer is difficult because little is known about its etiology. The main modifiable risk factors for pancreatic cancer include cigarette smoking and dietary factors. Information from molecular epidemiological study of pancreatic cancer is very limited. DNA adducts derived from exposure to polycyclic aromatic hydrocarbon, aromatic amines, and heterocyclic amines have been detected in human pancreatic tissues. DNA damages derived from oxidative stress and lipid peroxidation are also present in the pancreas. No study has demonstrated a main effect of carcinogen-metabolizing genes and DNA repair genes on the risk of pancreatic cancer thus far. However, significant effects of these genes have been observed among individuals with known carcinogen exposure, such as smoking. A number of environmental and lifestyle factors, such as smoking, alcohol, coffee consumption, and exposure to organochlorine or hydrocarbon solvent, have been associated with the frequency and spectrum of K-ras mutation in pancreatic tumors. Dietary folate intake and serum levels of folate have been associated with the risk of pancreatic cancer among male smokers. These findings demonstrate the potential of the molecular epidemiology approach in understanding the etiology of pancreatic cancer. Further efforts should be made to understand the interactive relationship between genetic and environmental factors in the etiology of pancreatic cancer, which will in turn be important in identifying the high-risk population for the primary prevention of this deadly disease. PMID- 12909736 TI - Chemoprevention for pancreatic cancer. AB - For a number of solid tumors, including pancreatic cancer, efforts aimed at disease prevention may be more successful than currently available anticancer treatments. While specific interventions are emerging to prevent breast, prostate, lung, and colorectal cancer, no trials of chemoprevention are being conducted in pancreatic cancer. Importantly, there are significant obstacles to the conduct of such research. However, preclinical and epidemiologic studies suggest that several drugs may have chemopreventive potential in pancreatic cancer. These include aspirin and other non-steroidal antiinflammatory drugs (NSAIDs), selective cyclooxygenase inhibitors, somatostatin analogs, selective estrogen receptor modulators (SERMs), and anti-androgenic agents. As the oncology community evaluates some of these agents in large chemoprevention trials for breast, colon, and prostate cancer, it may be found that pancreatic cancer prevention occurs as an unintended, but desirable consequence. Moreover, other general societal trends, such as smoking cessation and the widespread use of cholesterol-lowering agents and aspirin, could have a role in reducing the risk of pancreatic cancer, and in the future, may lead to a decrease in its incidence. PMID- 12909738 TI - A clinically relevant model of human pancreatic adenocarcinoma identifies patterns of metastasis associated with alterations of the TGF-beta/Smad4 signaling pathway. AB - Genetic alterations impacting the TGF-beta/Smad4 pathway are found in nearly all pancreatic adenocarcinomas, and recent reports have identified a relationship between DPC4/Smad4 expression and patient survival. In this study we use a clinically relevant animal model of pancreatic cancer to examine the impact of these genetic changes on the biology of pancreatic cancer. METHODS: Using high density oligonucleotide DNA microarray technology, a comprehensive examination of the components of the TGF-beta/Smad4 pathway was performed on three human pancreatic adenocarcinoma cell lines. The in vitro and in vivo growth characteristics of these cell lines was then compared. Finally, using a clinically relevant orthotopic xenograft model of pancreatic cancer, primary tumor growth and metastases were measured for pancreatic tumors derived from each cell line. RESULTS: Examination of the TGF-beta/Smad4 pathway components identified that these three cell lines possess molecular profiles consistent with approximately 90% of pancreatic adenocarcinoma tumors in patients. A significant discrepancy between in vitro and in vivo growth characteristics of each cell line was identified. When tumors from each cell line were established in nu/nu mice, each cell line exhibited distinct metastatic profiles. Data from these studies is consistent, with clinical observations concerning DPC4/Smad4 and patient outcome. CONCLUSION: Using an orthotopic model of tumor growth and metastasis identifies distinct metastatic profiles associated with molecular alterations of the TGF beta/Smad4 pathway and provides insight with regard to the biologic consequences of these changes. PMID- 12909735 TI - NF-kappaB in pancreatic cancer. AB - Although the genetic profile of pancreatic cancer is emerging as a result of much research, the role of specific genetic alterations that initiate tumorigenesis and produce its cardinal clinical features of locally aggressive growth, metastasis, and chemotherapy resistance remains unresolved. Recently, a number of studies have shown that the inhibition of constitutive NF-kappaB activation, one of the frequent molecular alterations in pancreatic cancer, inhibits tumorigenesis and metastasis. It also sensitizes pancreatic cancer cell lines to anticancer agent-induced apoptosis. Therefore because of the crucial role of NF kappaB in pancreatic cancer, it is a potential target for developing novel therapeutic strategies for the disease. In vivo and in vitro models that mimic the tumorigenic phenotypes in the appropriate histological and molecular concert would be very useful for confirming the suspected role of the pancreatic cancer signature genetic lesions and better understanding the molecular basis of this disease. PMID- 12909739 TI - Transgenic models of pancreatic cancer. PMID- 12909740 TI - Angiogenesis and antiangiogenic strategies in pancreatic cancer. AB - Despite numerous advances in the treatment of solid tumors, the prognosis of patients diagnosed with pancreatic cancer remains dismal. Results of both surgical and non-surgical treatment for pancreatic cancer have been extremely disappointing because of the tumors propensity to metastasize, failure of chemotherapy to achieve adequate levels within the tumor, and resistance of pancreatic cancer to cytotoxic agents. Current chemotherapeutic agents and radiation treatments rely on the rapidly dividing nature of tumor cells and are limited by their cytotoxic effects on normal cells. The need to exploit the difference between normal and malignant cells has resulted in an enormous amount of research into the process of tumor neovascularization. New agents are currently being developed that block tumor growth and metastasis through inhibition of angiogenesis. This article reviews the process of angiogenesis and antiangiogenic strategies with a special emphasis on pancreatic cancer. PMID- 12909737 TI - Experimental animal models of pancreatic carcinogenesis and metastasis. AB - Pancreatic cancer is a lethal disease characterized by early metastasis, local invasion, and resistance to conventional therapies. To understand its etiology and eventually make prevention of it possible and effective, appropriate carcinogenesis models will certainly help us understand the effects of environmental and genetic elements on pancreatic carcinogenesis. The development of new treatment strategies to control cancer metastasis is of immediate urgency. Fulfillment of this task relies on our knowledge of the cellular and molecular biology of pancreatic cancer metastasis and the availability of biologically and clinically relevant model systems. Many of the existing pancreatic cancer carcinogenesis and metastasis animal models are described in this review. The advantages and disadvantages of each model and their clinical implications are discussed, and special attention is focused on experimental therapeutic strategies targeting pancreatic cancer metastasis. PMID- 12909742 TI - A monoclinic polymorph of uranyl dinitrate trihydrate, [UO2(NO3)2(H2O)2].H2O. AB - Diaquadinitratouranyl(VI) monohydrate is monoclinic (space group P2(1)/c), in contrast to its triclinic polymorph. The main building block of the structure is the finite non-centrosymmetric [UO(2)(NO(3))(2)(H(2)O)(2)] cluster, which is a uranyl hexagonal bipyramid that shares two non-opposite equatorial edges with the nitrate triangles, such that the two water molecules are at neighbouring equatorial vertices. There is an interstitial water site in the structure, which is located between adjacent [UO(2)(NO(3))(2)(H(2)O)(2)] clusters. PMID- 12909741 TI - Novel gene therapy approaches to pancreatic cancer. AB - One of the most lethal human malignancies, pancreatic adenocarcinoma has remained a therapeutic challenge. Historically, the only curable treatment modalities available to patients with this disease have included pancreaticoduodenectomy with adjuvant chemoradiation. Few patients, however, have resectable disease at the time of presentation, and even for those who are offered this radical course of treatment, post-surgical survival is dismally short. Recently, however, advances in the understanding of the molecular biology of pancreatic cancer have enabled researchers to investigate novel gene therapy approaches to the treatment of this disease. In this paper, we review the common genetic mutations found in pancreatic adenocarcinomas, discuss strategies for the manipulation of genetic targets, and assess current progress in the field of gene therapy as it relates to pancreatic cancer. PMID- 12909743 TI - (K4Li4)Al8Ge8O32.8H2O: an Li+-exchanged potassium aluminogermanate with the zeolite gismondine (GIS) topology. AB - The title compound, lithium potassium dialuminium digermanium octaoxide dihydrate, (K,Li)-(Al,Ge)-GIS (GIS is gismondine), is the result of a 50% Li(+) exchange into the K-(Al,Ge)-GIS structure. The (K,Li)-(Al,Ge)-GIS structure was determined from a 4 x 4 x 2 micro m octahedral single crystal at the ESRF synchrotron X-ray source. The ion exchange results in a symmetry transformation from I2/a for K-(Al,Ge)-GIS to C2/c for (K,Li)-(Al,Ge)-GIS. The structural change is due to disordering of K(+) ions with Li(+) ions along the [001] channel and ordering of water molecules in the [101] channels. The distance between sites partially occupied by K(+) ions increases from 2.19 (3) A in K-(Al,Ge)-GIS to 2.94 (3) A in (K,Li)-(Al,Ge)-GIS. The Li(+) ions occupy positions along the twofold axis at the intersection of the eight-membered-ring channels in a twofold coordination with water molecules. For the four closest framework O(2-) anions, the Li.O distances are 3.87 (4) A. PMID- 12909745 TI - Two modifications of a KH2PO4.HF adduct. AB - The structures of two modifications, (I) and (II), of potassium dihydrogenphosphate-hydrofluoric acid (1/1), KH(2)PO(4).HF, were determined at 250 and 150 K, and at 292 and 150 K, respectively. Modifications (I) and (II) crystallize from stoichiometric aqueous solutions at 295 (1) and 308 (3) K, respectively. The H atoms were located clearly from the difference Fourier maps in each modification. The two modifications differ mainly in the arrangement of the dihydrogenphosphate anions, i.e. (I) contains looped dimeric and tetrameric units of the dihydrogenphosphate ions, whereas (II) contains two types of looped tetrameric unit. In addition, both structures contain a very short F-H.O hydrogen bond (2.38-2.40 A). The K(+) ions are coordinated by O and F atoms, with similar K.O and K.F distances in both modifications. PMID- 12909744 TI - Manganese antimony diselenide iodide, MnSbSe2I. AB - MnSbSe(2)I contains layers parallel to the bc plane that consist of edge- and corner-sharing MnSe(6/2) and MnSe(2/2)I(4/2) octahedra. Sb atoms are located between these layers and form SbSe(3) trigonal pyramids. Owing to the off-center placement of the Sb atom and the inequivalence of the two crystallographically independent Mn atoms, the monoclinic MnSbSe(2)I structure is a distorted variant of the orthorhombic UFeS(3) structure type. PMID- 12909746 TI - A new cubic form of caesium hexaaquamagnesium phosphate, Cs[Mg(H2O)6](PO4). AB - A new cubic form (space group F-43m) of the title compound has been found which is isostructural with the analogous arsenate. [Mg(H(2)O)(6)](2+) cations and phosphate anions are connected by hydrogen bonds, forming a sphalerite-like three dimensional framework. PMID- 12909747 TI - Ca2MgWO6 from neutron and X-ray powder data. AB - The room-temperature structure of the B-site-ordered complex perovskite dicalcium magnesium tungstate, Ca(2)MgWO(6), has been determined by simultaneous Rietveld refinement of neutron and X-ray powder diffraction patterns. Ca(2)MgWO(6) is characterized by B-site ordering and an a(-)a(-)c(+)-type BO(6) octahedral tilt mechanism. PMID- 12909749 TI - A new Na/Mg inverse crown ether. AB - A 'missing' member of the inverse crown ether family, namely mu(4)-oxo tetrakis(mu-2,2,6,6-tetramethylpiperidinido)dimagnesium(II)disodium(I), [Na(2)Mg(2)O(C(9)H(18)N)(4)], has been synthesized by blocking the alternative aromatic metallation route via the use of sterically hindered 1,3,5-mesitylene as a solvent. [Na(2)Mg(2)O(NR(2))(4)] (NR(2) is 2,2,6,6-tetramethylpiperidinide) is shown to form a cationic planar eight-membered ring with alternating metal and N atoms, which captures at its core an oxide guest that lies on an inversion centre [principal dimensions: Na-O = 2.2405 (11) A, Na-N = 2.445 (3) and 2.572 (3) A, Mg O = 1.8673 (9) A, and Mg-N = 2.032 (2) and 2.063 (2) A]. PMID- 12909748 TI - [2,6-Bis(1H-benzimidazol-2-yl-kappaN3)pyridine-kappaN](dimethylformamide kappaO)(thiosulfato-kappa2O,S)nickel(II) monohydrate. AB - The structure of the title compound, [Ni(ths)(bbip)(dmf)].H(2)O [ths is thiosulfate, S(2)O(3); bbip is 2,6-bis(1H-benzimidazol-2-yl)pyridine, C(21)H(13)N(5); and dmf is dimethylformamide, C(3)H(7)NO], is monomeric, with the nickel ion octahedrally surrounded by an N,N',N"-tridentate bbip molecule, an S,O bidentate ths molecule and an O-monodentate dmf molecule. The H atoms of the hydration water molecule and the amino groups of bbip are involved in hydrogen bonding and determine a spatial organization of broad layers parallel to (001), which are connected by weak interactions. PMID- 12909750 TI - A novel cobalt(II) coordination polymer containing the fumarate anion and o phenanthroline. AB - The self-assembly of three crystallographically distinct fumarate ions, two unique cobalt(II) ions and two unique o-phenanthroline molecules results in a two dimensional polymeric structure with the formula [Co(2)(C(4)H(2)O(4))(2)(C(12)H(8)N(2))(2)](n), namely di-mu-fumatato-bis(o phenanthroline)dicobalt(II). The Co atoms are at the nodes of a two-dimensional array linked by coordinated fumarate ligands. Each Co atom is coordinated in a distorted octahedral manner to four fumarate O atoms and two N atoms from the chelating phenanthroline ligands. PMID- 12909751 TI - (2,2'-Diamino-4,4'-bi-1,3-thiazole-kappa2N3,N3')(oxydiacetato kappa3O,O',O")copper(II) monohydrate. AB - The title compound, [Cu(C(4)H(4)O(5))(C(6)H(6)N(4)S(2))].H(2)O, displays a square pyramidal coordination geometry. The tridentate oxydiacetate dianion chelates the Cu(II) atom in the facial mode. The large difference [0.487 (4) A] between the longest Cu-O distance in the basal plane and that in the apical direction correlates with the small displacement of the Cu(II) atom [0.0576 (13) A] from the basal plane towards the apex of the square pyramid. The intermolecular hydrogen-bonding network results in a closely overlapped arrangement of the coordination basal plane and the thiazole ring of a neighboring molecule. PMID- 12909752 TI - cis-[1,4-Bis(diphenylphosphino)butane](mu-tetrathiotungstato)palladium(II) N,N' dimethylformamide hemisolvate hemihydrate. AB - In the title compound, [1,4-bis(diphenylphosphino)butane-2kappa(2)P,P']di-mu-thio 1:2kappa(4)S-dithio-1kappa(2)S-palladium(II)tungsten(VI) N,N'-dimethylformamide hemisolvate hemihydrate, [PdWS(4)(C(28)H(28)P(2))].0.5C(3)H(7)NO.0.5H(2)O, the Pd atom is coordinated by two S atoms from the distorted-tetrahedral [WS(4)](2-) anion and two P atoms from the dppb molecule [dppb is 1,4 bis(diphenylphosphino)butane] in an approximately square-planar configuration. A puckered seven-membered ring is formed by the Pd atom and the dppb ligand. PMID- 12909753 TI - A one-dimensional zigzag coordination polymer of diaqua(pyridine-2,3 dicarboxylato)cobalt(II). AB - The asymmetric unit of the title one-dimensional coordination polymer, catena poly[[mu-pyridine-2,3-dicarboxylato-1kappaO:2kappa(2)N,O'-bis[diaquacobalt(II)]] mu-pyridine-2,3-dicarboxylato-1kappa(2)N,O:2kappaO':1'kappaO'], [Co(C(7)H(3)NO(4))(H(2)O)(2)](n), is composed of a cobalt(II) ion, a pyridine-2,3 dicarboxylate dianion and two water molecules. The polymer has a zigzag structure consisting of a chain of edge-fused rings, and the polymer chains are linked by O H.O hydrogen bonds into a three-dimensional framework. PMID- 12909754 TI - Potassium N-(6-amino-3,4-dihydro-3-methyl-5-nitroso-4-oxopyrimidin-2-yl)-(S) aspartate N-(6-amino-3,4-dihydro-3-methyl-5-nitroso-4-oxopyrimidin-2-yl)-(S) aspartic acid 4.88-hydrate: a two-dimensional coordination polymer. AB - In the title compound, K(+).C(9)H(10)N(5)O(6)(-).C(9)H(11)N(5)O(6).4.88H(2)O, the cations are eight-coordinate. There is a one-dimensional coordination-polymer substructure, built from K and O atoms only, in the form of chains of distorted confacial octahedra, and these are linked into sheets by the neutral organic ligand. The sheets are linked into a single framework by a combination of N-H.O and O-H.O hydrogen bonds. PMID- 12909755 TI - Hexaaquairon(II) dipicrate dihydrate. AB - In the crystal structure of the title compound, [Fe(H(2)O)(6)](C(6)H(2)N(3)O(7))(2).2H(2)O, the centrosymmetric cationic iron complexes and picrate anions form separate stacks extending along the b axis. No picrate species ligate to the metal cation. Picrate ions are linked to one another in the stack via short intermolecular C.C contacts of 3.083 (4) and 3.055 (4) A. Variable-temperature X-ray diffraction measurements performed between room temperature and 93 K showed a linear decrease of the lattice parameters, suggesting that there is no phase transition. PMID- 12909756 TI - Tricarbonyl(eta6-chlorobenzene)chromium. AB - The title compound, [Cr(C(6)H(5)Cl)(CO)(3)], is the first group 6 tricarbonyl eta(6)-monohaloarene compound to be structurally characterized. It adopts a classic piano-stool structure, with the Cr(CO)(3) tripod assuming a syn-eclipsed conformation relative to the arene ring (varphi = 2.0 degrees ). The extended structure is dominated by intermolecular pi.H interactions (H.ring centroid = 2.94 A) and non-classical hydrogen bonds between carbonyl and arene moieties (O.H = 2.50-2.58 A). PMID- 12909757 TI - Potassium dicyanoargentate(I) 1,4,7,10,13,16-hexaoxacyclooctadecane. AB - The crystal structure of the title compound, K[Ag(CN)(2)].C(12)H(24)O(6), conventionally denoted K(18-crown-6)Ag(CN)(2), where 18-crown-6 is 1,4,7,10,13,16 hexaoxacyclooctadecane, is characterized by closely packed linear chains formed by the coordination of the nitrile N atoms of the [Ag(CN)(2)](-) anions to the K(+) cations. The K atoms lie on centers of inversion and are additionally bound to the six O atoms of the crown ether. PMID- 12909758 TI - Ferrocene compounds. XXXIX. 1-Ferrocenylisochromane. AB - In the title compound, [Fe(C(5)H(5))(C(14)H(13)O)], the plane of the heterocyclic ring is almost perpendicular to the plane of the substituted cyclopentadienyl ring, and the heterocyclic ring adopts a half-chair conformation. The conformation of the nearly parallel cyclopentadienyl (Cp) rings [the dihedral angle between their planes is 2.7 (1) degrees ] is almost halfway between eclipsed and staggered, and the rings are mutually twisted by about 19.4 (2) degrees (mean value). The mean lengths of the C-C bonds in the substituted and unsubstituted cyclopentadienyl ring are 1.420 (2) and 1.406 (3) A, respectively, and the Fe-C distances range from 2.029 (2) to 2.051 (2) A. The phenyl and unsubstituted cyclopentadienyl rings are involved in C-H.pi interactions, with intermolecular H.centroid distances of 2.85 and 3.14 A for C-H.pi(Ph), and 2.88 A for C-H.pi(Cp). In two of these interactions, the C-H bond points towards one of the ring bonds rather than towards the ring centroid. In the crystal structure, the C-H.pi interactions connect the molecules into a three-dimensional framework. PMID- 12909759 TI - Bis[1-(p-chlorophenyl)-5-isopropylbiguanide-kappa2N2,N4]nickel(II) dichloride dimethylformamide solvate. AB - The asymmetric unit of the title compound, [Ni(C(11)H(16)ClN(5))(2)]Cl(2).C(3)H(7)NO, contains one monomeric nickel(II) complex cation, two Cl(-) anions and one dimethylformamide solvent molecule. The Ni atom is coordinated to each of two 1-(p-chlorophenyl)-5-isopropylbiguanide (proguanil) ligands via two N atoms. The complex exhibits a square-planar coordination, with the Ni atom lying 0.021 (2) A out of the basal plane. The crystal packing is characterized by several hydrogen bonds. PMID- 12909760 TI - [Complexation du 2-[(5-methylpyrazolyl)methyle]benzimidazole par les chlorures de cuivre et de cadmium]. AB - The structures of dichloro[2-[(5-methyl-1H-pyrazol-3-yl-kappaN(2))methyl]-1H-1,3 benzimidazole-kappaN(3)]copper(II), [CuCl(2)(C(12)H(12)N(4))], and di-mu-chloro bis(chloro[2-[(5-methyl-1H-pyrazol-3-yl-kappaN(2))methyl]-1H-1,3-benzimidazole kappaN(3)]cadmium(II)), [Cd(2)Cl(4)(C(12)H(12)N(4))(2)], show that these compounds have the structural formula [ML(Cl)(2)](n), where L is 2-[(5 methylpyrazolyl)methyl]benzimidazole. When M is copper, the complex is a monomer (n = 1), with a tetrahedral coordination for the Cu atom. When M is cadmium (n = 2), the complex lies about an inversion centre giving rise to a centrosymmetric dimer in which the Cd atoms are bridged by two chloride ions and are pentacoordinated. PMID- 12909761 TI - Polymeric diaquabis[mu4-1,3,5-benzenetricarboxylato(3-)](mu-4,4' bipyridine)trizinc(II). AB - The solvothermal reaction of zinc(II) acetate with 1,3,5-benzenetricarboxylic acid and 4,4'-bipyridine in 1-butanol and acetic acid solution gives the title complex, [Zn(3)(C(9)H(3)O(6))(2)(C(10)H(8)N(2))(H(2)O)(2)](n). There are two different coordination environments for the Zn atom, namely ZnNO(2)(H(2)O) and ZnO(4); in the latter case, the Zn atom lies on a twofold symmetry axis. Carboxylate-bridged Zn(3)(O(2)C)(6) clusters are connected through 1,3,5 benzenetricarboxylate and 4,4'-bipyridine ligands (which lie about inversion centers), yielding a three-dimensional framework. PMID- 12909762 TI - Unusual molecular conformation in dissymmetric propylene-linker compounds containing pyrazolo[3,4-d]pyrimidine and phthalimide moieties. AB - The crystal structure of 4,6-bis(methylsulfanyl)-1-phthalimidopropyl-1H pyrazolo[3,4-d]pyrimidine, C(18)H(17)N(5)O(2)S(2), (VI), reveals an unusual folded conformation due to an apparent intramolecular C-H.pi interaction between the 6-methylsulfanyl and phenyl groups. However, the closely related compound 6 methylsulfanyl-1-phthalimidopropyl-4-(pyrrolidin-1-yl)-1H-pyrazolo[3,4 d]pyrimidine, C(21)H(22)N(6)O(2)S, (VII), exhibits a fully extended structure, devoid of any intramolecular C-H.pi or pi-pi interactions. The crystal packing of both molecules involves intermolecular stacking interactions due to aromatic pi pi interactions. In addition, (VI) exhibits intermolecular C-H.O hydrogen bonding and (VII) exhibits dimerization of the molecules through intermolecular C-H.N hydrogen bonding. PMID- 12909763 TI - The tetrapeptide Z-Leu-Aib-Pro-Val-OBg monohydrate. AB - The intramolecular hydrogen-bonding pattern of Z-Leu-Aib-Pro-Val-OBg monohydrate [(N-benzhydrylamino)carbonylmethyl N-benzyloxycarbonyl-alpha aminoisobutyrylprolylvalinate monohydrate], C(43)H(55)N(5)O(8).H(2)O, is unusual for a tetrapeptide because, in addition to a 1-->4 hydrogen bond, a second hydrogen bond of the type 1-->5 is formed. This folding reflects the intramolecular hydrogen-bonding pattern that this amino acid sequence adopts in the naturally occurring peptaibol alamethicin. PMID- 12909764 TI - Samaderin B and C from Samadera indica. AB - Samaderin B, or (1R,2S,5R,5aR,7aS,11S,11aS,11bR,14S)-1,7,7a,11,11a,11b-hexahydro 1,11-dihydroxy-8,11a,14-trimethyl-2H-5a,2,5-(methanoxymetheno)naphth[1,2 d]oxepine-4,6,10(5H)-trione, C(19)H(22)O(7), and samaderin C, or (1R,2S,5R,5aR,7aS,10S,11S,11aS,11bR,14S)-7,7a,10,11,11a,11b-hexahydro-1,10,11 trihydroxy-8,11a,14-trimethyl-2H-5a,2,5-(methanoxymetheno)naphth[1,2-d]oxepine 4,6(1H,5H)-dione, C(19)H(24)O(7), were isolated from the seed kernels of Samadera indica and were shown to exhibit antifeedant activity against Spodoptera litura third-instar larvae. The replacement of the carbonyl group in samaderin B by a hydroxy group in samaderin C causes conformational changes at the substitution site, but the overall conformation is not affected; however, the compounds pack differently in the crystal lattice. PMID- 12909765 TI - 4,5-Dihydro-3-methyl-5-(4-methylphenyl)-1H-pyrazole-1-carboxamide. AB - The reaction between 4-(4-methylphenyl)but-3-en-2-one and aminoguanidine produced an unexpected product of formula C(12)H(15)N(3)O, consisting of a carboxamide moiety joined to a substituted pyrazoline ring at one of the N atoms. The pyrazoline ring adopts a flat-envelope conformation and the substituted phenyl ring is oriented almost perpendicular to the heterocycle. The carbonyl O atom has partial anionic character as a result of the transfer of pi density from the two adjacent sp(2) N atoms and is involved in an intermolecular hydrogen bond with the amide group. PMID- 12909766 TI - 2-(2-Naphthyloxy)acetate derivatives. I. A new class of antiamnesic agents. AB - The title compounds 1-(2-naphthyloxymethylcarbonyl)piperidine, C(17)H(19)NO(2), (I), and 3-methyl-1-(2-naphthyloxymethylcarbonyl)piperidine, C(18)H(21)NO(2), (II), are potential antiamnesics. In (II), the methyl-substituted piperidine ring is disordered over two conformations. The piperidine ring has a chair conformation in both compounds. In (I), the molecules are linked by weak intermolecular C-H.O interactions to give networks represented by C(4), C(6) and R(4)(4)(18) graph-set motifs, while in (II), weak intermolecular C-H.O interactions generate R(1)(2)(5), C(4) and C(7) graph-set motifs. The dihedral angle between the naphthalene moiety and the piperidine ring is 33.83 (7) degrees in (I), while it is 31.78 (11) and 19.38 (19) degrees for the major and minor conformations, respectively, in (II). PMID- 12909767 TI - 1,3-Bis(4-nitrophenyl)triazene. AB - The structure of the title compound, C(12)H(9)N(5)O(4), reveals an almost planar molecule (r.m.s. deviation = 0.061 A), in which the interplanar angle between the phenyl rings is 5.7 (1) degrees and the largest interplanar angle is that between the phenyl ring and the nitro group of one of the 4-nitrophenyl substituents [8.8 (3) degrees ]. The observed molecular conformation suggests a delocalization of pi-electrons extended over the diazoamine group and the terminal aryl substituents. Intermolecular N-H.O interactions between the twofold screw-related molecules give rise to helical chains along the [010] direction. Intermolecular C H.O interactions then generate sheets of molecules in the (10-1) plane, and these sheets are held together by N.C and O.O pi-pi interactions. PMID- 12909768 TI - Simvastatin. AB - Simvastatin, or (1S,3R,7S,8S,8aR)-8-[2-[(2R,4R)-4-hydroxy-6-oxo-3,4,5,6 tetrahydro-2H-pyran-2-yl]ethyl]-3,7-dimethyl-1,2,3,7,8,8a-hexahydronaphthalen-1 yl 2,2-dimethylbutanoate, C(25)H(38)O(5), is almost isostructural with lovastatin, and the general conformational features are closely related to those of other reported crystal structures of statins. The only hydrogen bond present facilitates the formation of infinite chains of molecules along the b axis. PMID- 12909769 TI - 11-Hydroxy-3,3-dimethyl-7,12-dioxo-3,4,6,6a,7,12,12a,12b octahydrobenz[a]anthracen-1-yl acetate. AB - The Diels-Alder reaction between 5-hydroxy-1,4-naphthoquinone and 5,5-dimethyl-3 vinyl-1,2-cyclohexadienyl acetate by endo addition gives the title compound, C(22)H(22)O(5), in 68% yield. This racemic diastereoisomer has the opposite regiochemistry to ochromycinone analogues produced previously and may allow access to a new type of anticancer-active saquayamycin analogue. PMID- 12909770 TI - Tetraphenylphosphonium 4-hydroxy-2,2,6,6-tetramethylpiperidinyloxy-4-sulfonate. AB - The title compound, C(24)H(20)P(+).C(9)H(17)NO(5)S(-), consists of an organic monovalent cation and an organic monovalent anion, the latter being derived from the TEMPO radical (TEMPO is 2,2,6,6-tetramethylpiperidin-1-oxyl). Two inversion related anions interact via two -O-H.O-S- hydrogen bonds, forming a dimer in which there are no short contacts between the spin centres (-N-O) of the TEMPO(OH)SO(3)(-) anions. Furthermore, no significant magnetic interaction is observed between the dimers because the dimer is surrounded by cations. These results are consistent with the paramagnetic behaviour of the title salt. PMID- 12909771 TI - Cis,cis,cis-1,2,4,5-cyclohexanetetracarboxylic acid and its dianhydride. AB - cis,cis,cis-1,2,4,5-Cyclohexanetetracarboxylic acid, C(10)H(12)O(8), (I), contains a mirror plane and the cyclohexane ring exhibits a chair conformation. Two crystallographically independent hydrogen bonds form R(2)(2)(14), R(2)(2)(16) and R(4)(4)(16) ring motifs, and propagation of these two hydrogen bonds along the c and b axes generates C(2)(2)(16) and C(2)(2)(7) chains. cis,cis,cis-1,2:4,5 Cyclohexanetetracarboxylic dianhydride, C(10)H(8)O(6), (II), was prepared by the reaction of (I) with acetic anhydride. The cyclohexane ring of (II) exhibits a boat conformation and the dihedral angle between the two anhydro rings is 117.5 (1) degrees. PMID- 12909772 TI - Diiododurene: four centrosymmetric molecules in general positions. AB - Diiododurene (1,4-diodo-2,3,5,6-tetramethylbenzene), C(10)H(12)I(2), packs with four molecules in the asymmetric unit. All four of these moleules violate Kitaigorodsky's suggestion that molecules with centers of symmetry will lie on crystallographic centers of symmetry. There is 5.6% disorder at one of the sites. Most of the I atoms are in contact with other I atoms, but only six of the I.I contacts are shorter than 4.2 A. Of these six contacts, one set of three contacts forms a triangular set in which all of the I.I distances are less than 3.9 A. PMID- 12909773 TI - 7-Bromoquinolin-8-ol. AB - Structure analysis of the title compound, C(9)H(6)BrNO, has established that bromination of an 8-hydroxyquinoline derivative occurred in the 7-position. Intermolecular and weak intramolecular O-H.N hydrogen bonds are present, the former causing the molecules to pack as hydrogen-bonded dimers in the solid state. PMID- 12909774 TI - The low-temperature phase of 1,3-dibromo-2,4,6-trimethylbenzene: a single-crystal neutron diffraction study at 120 and 14 K. AB - In the low-temperature phase of dibromomesitylene (1,3-dibromo-2,4,6 trimethylbenzene), C(9)H(10)Br(2), the molecule deviates significantly from the C(3h) molecular symmetry encountered in tribromomesitylene (1,3,5-tribromo-2,4,6 trimethylbenzene), even for the endocyclic bond angles. An apparent C(2v) molecular symmetry is observed. The angle between the normal to the molecular plane and the normal to the (100) plane is approximately 20 degrees. The overall displacement was analysed at 120 K with rigid-body-motion tensor analysis. The methyl group located intermediate between the two Br atoms is rotationally disordered at both temperatures. This disorder was treated using two different approaches at 14 K, viz. the conventional split-atom model and a model using the special annular shapes of the atomic displacement parameters that are available in CRYSTALS [Watkin, Prout, Carruthers & Betteridge (1999). Issue 11. Chemical Crystallography Laboratory, Oxford, England], but only through the latter approach at 120 K. The disorder locally breaks the C(2v) molecular symmetry at 14 K only. Intra- and intermolecular contacts are described and discussed in relation to this methyl-group disorder. The bidimensional pseudo-hexagonal structural topology of trihalogenomesitylenes is altered in dibromomesitylene insofar as the (100) molecular layers are undulated and are not coplanar as a result of an alternating tilt angle of approximately 34 degrees propagating along the [011] and [0-11] directions between successive antiferroelectric molecular columns oriented roughly along the a axis. PMID- 12909775 TI - 17beta-Ethoxy-3-methoxy-8-isoestra-1,3,5(10)-triene. AB - The conformation of the crystal of 17beta-ethoxy-3-methoxy-8-isoestra-1,3,5(10) triene, C(21)H(30)O(2), (I), has been established and compared with the molecular structure of a typical steroid estrogen 8-iso-analogue, (II). Calculations of distances separating some of the H-atom pairs in (I) and (II) by molecular mechanical and semi-empirical methods revealed the similarity of the values to the H.H distances obtained from X-ray analysis. PMID- 12909776 TI - Benzylation and nitrosation of 4-amino-2-(methylsulfanyl)pyrimidin-6(1H)-one: two P2(1)/c polymorphs of 4-amino-1-benzyl-2-(methylsulfanyl)pyrimidin-6(1H)-one with Z' = 1 and 2, 4-amino-6-benzyloxy-2-(methylsulfanyl)pyrimidine and 4-amino-1 benzyl-2-(methylsulfanyl)-5-nitrosopyrimidin-6(1H)-one all give different hydrogen-bonded supramolecular structures. AB - 4-Amino-1-benzyl-2-(methylsulfanyl)pyrimidin-6(1H)-one, C(12)H(13)N(3)OS, crystallizes in two polymorphic forms, both having space group P2(1)/c, with Z' = 1 for form (I) and Z' = 2 for form (II). In (I), the molecules are linked by a single N-H.O hydrogen bond into simple C(6) chains, which are themselves linked into sheets by aromatic pi-pi interactions, while in (II), chains of edge-fused R(4)(2)(8) and R(4)(4)(24) rings are generated by four distinct N-H.O hydrogen bonds. In 4-amino-1-benzyl-2-(methylsulfanyl)-5-nitrosopyrimidin-6(1H)-one, C(12)H(12)N(4)O(2)S, (III), where Z' = 2, two independent three-centre N-H.(N,O) hydrogen bonds generate a C(5)C(6)[R(1)(2)(5)] chain of rings. In 4-amino-6 benzyloxy-2-(methylsulfanyl)pyrimidine, C(12)H(13)N(3)OS, (IV), which is isomeric with (I) and (II), a combination of N-H.N and N-H.O hydrogen bonds generates a sheet of alternating R(2)(2)(8) and R(6)(6)(28) rings. PMID- 12909777 TI - 1,4,8,11-Tetraazoniacyclotetradecane tetrakis(phenylphosphonate) dihydrate forms hydrogen-bonded sheets. AB - In the title compound, C(10)H(28)N(4)(4+).4C(6)H(6)O(3)P(-).2H(2)O, the cation lies across a centre of inversion in space group P2(1)/n. The two independent anions and the water molecule are linked by four O-H.O hydrogen bonds [H.O = 1.74 2.15 (3) A, O.O = 2.5482 (14)-3.0438 (16) A and O-H.O = 160-176 (2) degrees ] into sheets containing equal numbers of centrosymmetric R(6)(6)(16) and R(10)(10)(36) rings. The cation lies at the centre of the R(10)(10)(36) ring and is linked to it by eight N-H.O hydrogen bonds [H.O = 1.77-1.98 A, N.O = 2.6756 (15)-2.8168 (16) A and N-H.O = 151-169 degrees ]. PMID- 12909778 TI - 4,4'-Trimethylenedipyridinium bis[carboxymethylphosphonate(1-)]: a three dimensional framework structure built from O-H...O, N-H...O and C-H...O hydrogen bonds. AB - In the title compound, C(13)H(16)N(2)(2+).2C(2)H(4)O(5)P(-), the cation lies across a twofold rotation axis in space group Fdd2. The anions are linked into molecular ladders by two O-H.O hydrogen bonds [H.O = 1.73 and 1.77 A, O.O = 2.538 (2) and 2.598 (3) A, and O-H.O = 160 and 170 degrees ], these ladders are linked into sheets by a single type of N-H.O hydrogen bond [H.O = 1.75 A, N.O = 2.624 (3) A and N-H.O = 171 degrees ] and the sheets are linked into a three dimensional framework by a single type of C-H.O hydrogen bond [H.O = 2.48 A, C.O = 3.419 (4) A and C-H.O = 167 degrees ]. PMID- 12909779 TI - 2-(2-Naphthyloxy)acetate derivatives. II. A new class of antiamnesic agents. AB - The title compounds, 4-(2-naphthyloxymethylcarbonyl)morpholine, C(16)H(17)NO(3), (I), and 4-methyl-1-(2-naphthyloxymethylcarbonyl)piperazine, C(17)H(20)N(2)O(2), (II), are potential antiamnesics. The morpholine ring in (I) and the piperazine ring in (II) adopt chair conformations. In (I), the molecules are linked by weak intermolecular C-H.O interactions into chains that have a graph-set motif of C(10), while in (II), the molecules are linked by weak intermolecular C-H.O interactions that generate two C(7) graph-set motifs. The dihedral angle between the naphthalene moiety and the best plane through the morpholine ring is 20.62 (4) degrees in (I), while the naphthalene moiety is oriented nearly perpendicular to the mean plane of the piperazine ring in (II). PMID- 12909780 TI - 4-[N,N-Bis(2-cyanoethyl)amino]pyridine. AB - The title compound, 3,3'-(4-pyridylimino)dipropanenitrile, C(11)H(12)N(4), has a twofold axis and consists of a pyridine ring head and two cyanoethyl tails, the three groups being linked by an N atom. The planar geometry around the amino N atom suggests conjugation with the pi-system of the pyridine ring. The molecules are stacked in a layer structure via relatively weak to very weak intermolecular C-H.pi and C-H.N hydrogen-bond interactions. PMID- 12909781 TI - An intermediate in a new synthesis approach to beta-substituted beta hydroxyaspartame. AB - The crystal and molecular structure of 1-tert-butyl 4-ethyl (2'R,3'R,5'R,2S,3S)-3 bromomethyl-3-hydroxy-2-[(2'-hydroxy-2',6',6'-trimethylbicyclo[3.1.1]hept-3' ylidene)amino]succinate, C(21)H(34)BrNO(6), is presented. This compound is an intermediate in the new synthetic route to beta-substituted beta hydroxyaspartates, which are blockers of glutamate transport. PMID- 12909782 TI - C-H...O and C-H...pi interactions in 1-acetyl-4-(p-chlorobenzylideneamino)-3 ethyl-4,5-dihydro-1H-1,2,4-triazol-5-one. AB - The title compound, C(13)H(13)ClN(4)O(2), contains both a phenyl and a triazole ring, both of which are approximately coplanar with the entire molecule. The triazole ring has substituents at the 1-, 2- and 4-positions. Intramolecular C H.O and C-H.N interactions, together with intermolecular C-H.O and C-H.pi interactions, help to stabilize the structure. PMID- 12909783 TI - (1R,3S)-1-Monoamidocamphoric acid. AB - The title compound, (1S,3R)-3-carbamoyl-2,2,3-trimethylcyclopentane-1-carboxylic acid, C(10)H(17)NO(3), was synthesized and characterized by IR, EA, ES-MS (electrospray ionization mass spectra), (1)H NMR, (13)C NMR and X-ray diffraction techniques. The two independent molecules form a two-dimensional network via O H.O and N-H.O hydrogen-bonding interactions between their carboxylic acid and carbamoyl groups. PMID- 12909784 TI - Hydrogen-bond networks in tris(4-hydroxyphenyl)methane and its 1:1 molecular complex with 4,4'-bipyridine. AB - In tris(4-hydroxyphenyl)methane (or 4,4',4"-methanetriyltriphenol), C(19)H(16)O(3), molecules are connected by O-H.O hydrogen bonds [O.O = 2.662 (2) and 2.648 (2) A] into two-dimensional square networks that are twofold interpenetrated. In tris(4-hydroxyphenyl)methane-4,4'-bipyridine (1/1), C(19)H(16)O(3).C(10)H(8)N(2), trisphenol molecules form rectangular networks via O-H.O [O.O = 2.694 (3) A] and C-H.O [C.O = 3.384 (3) A] hydrogen bonds. Bipyridine molecules hydrogen bonded to phenol moieties [O.N = 2.622 (3) and 2.764 (3) A] fill the voids to complete the structure. PMID- 12909785 TI - aka primary nursing. PMID- 12909786 TI - Qualitative analysis of telehomecare nursing activities. PMID- 12909787 TI - Grievance tracking: targeting an improvement process. PMID- 12909788 TI - Nosocomial pressure ulcer prevention: a successful project. PMID- 12909789 TI - Personal characteristics and job satisfaction among nurses in Lebanon. AB - OBJECTIVE: This study investigates the level of satisfaction of Lebanese nurses in their job and the influence of their personal characteristics. BACKGROUND: Given the current difficulties experienced by Lebanese hospitals in recruiting and retaining a sufficient number of nurses, the need to understand the reasons of nurses' dissatisfaction became urgent. Moreover, satisfaction at work is essentially a personal experience also affected by cultural factors. Therefore, it was necessary to study the links between personal characteristics and nurses' dissatisfaction. METHODS: The study included 421 registered nurses. A modified version of Measure of Job Satisfaction, developed by Taynor and Wade, was used to assess the effect of the personal characteristics, namely educational level, age, years of work experience, position, and marital and parental status, on 5 dimensions of satisfaction: personal satisfaction, workload, professional support, pay and prospect, and training. RESULTS: The findings suggest that personal characteristics have important influences on nurses' job perceptions. University graduate nurses reported more dissatisfaction with the quality of supervision and with respect and treatment they receive from their superiors. Nurses younger than 30 years and the technically trained were more dissatisfied with the available opportunities to attend continuing education courses. Results of staff nurses and unmarried nurses showed trends of more dissatisfaction than the married and nurses of higher positions. Moreover, the whole sample perceived that nursing provided a high level of personal satisfaction, but nurses were most dissatisfied with salary and lack of prospects for promotion. CONCLUSION: The results indicate the importance of personal characteristics on nurses' retention. Furthermore, intrinsic factors related to the nature and experience of nursing are more job satisfying than extrinsic factors. PMID- 12909790 TI - Providing staff feedback to nurse managers using internal resources. AB - In many regions of the country, hospitals are faced with increasingly serious nursing shortages. Many hospitals have developed incentive packages to recruit nurses. However, the literature clearly demonstrates that the nurses' work environments must be addressed if nurses are to be retained. The authors created a questionnaire to measure registered nurses' attitudes toward their work environments. Information from this questionnaire can provide timely feedback and enable nurse managers to identify and correct organizational factors that could lead to turnover. The questionnaire takes fewer than 10 minutes for each nurse to complete, and nurse managers can easily calculate the results. The questionnaire can be modified for use in other healthcare facilities. PMID- 12909791 TI - Translating psychosocial research findings into practice guidelines. AB - Evidence-based practice guidelines are increasingly used by healthcare professionals to guide patient care and effect positive patient outcomes. These guidelines are usually based on laboratory-based parameters and lack the psychosocial dimensions of patient care. The authors describe the process used by a hospital-based interdisciplinary team to successfully develop evidence-based psychosocial guidelines designed to foster hopefulness in pediatric patients with cancer, their families, and their healthcare providers. From these guidelines, 4 clinical care projects were developed and are described in this article. PMID- 12909792 TI - Vision 2020, part 2: Educational preparation for the future nurse manager. AB - The demand for knowledgeable and skilled nursing leaders at the first, middle, and executive levels of management in healthcare organizations drove a multiphased project concerning the types of nurse managers that will be needed in the future and their educational needs. In Part 1 (June 2003), national nurse leaders identified the increased need for knowledgeable and skilled nurses in first-line, mid-level, and executive management positions in the healthcare delivery system. This article describes the second Vision 2020 survey of nurse managers in Massachusetts on the current and future demand for nurse managers and their academic preparation and continuing education needs. It concludes with a model curriculum revision for a master's program in nursing management for the public urban university in Boston. PMID- 12909793 TI - Workplace empowerment and magnet hospital characteristics: making the link. AB - OBJECTIVE: To test a theoretical model linking nurses' perceptions of workplace empowerment, magnet hospital characteristics, and job satisfaction in 3 independent studies of nurses in different work settings. BACKGROUND: Strategies proposed in Kanter's structural empowerment theory have the potential to result in work environments that are described in terms of magnet hospital characteristics. Identifying factors that contribute to work conditions that attract and retain highly qualified committed nurses, such as those found in magnet hospitals, that can be put in place by nursing administrators is extremely important for work redesign to promote professional nursing practice. METHODS: Secondary analyses of data from 3 studies were conducted--2 of staff nurses and 1 with acute care nurse practitioners working in Ontario, Canada. The Conditions of Work Effectiveness Questionnaire-II, the NWI-R, and measures of job satisfaction were used to measure the major study variables.RESULTS The results of all 3 studies support the hypothesized relationships between structural empowerment and the magnet hospital characteristics of autonomy, control over practice environment, and positive nurse-physician relationships. The combination of access to empowering work conditions and magnet hospital characteristics was significantly predictive of nurses' satisfaction with their jobs. CONCLUSIONS/IMPLICATIONS: These findings suggest that nursing leaders' efforts to create empowering work environments can influence nurses' ability to practice in a professional manner, ensuring excellent patient care quality and positive organizational outcomes. PMID- 12909794 TI - Preface. Nursing informatics. A foundation for nursing professionalism. PMID- 12909795 TI - Nursing informatics: the key to unlocking contemporary nursing practice. AB - Nursing informatics is a relatively new nursing specialty. Recognized by the American Nurses' Association in 1992, this field within nursing has grown exponentially. Once the purview of highly specialized individuals, nursing informatics has now crept into all dimensions of nursing, from domain of advanced nurse practitioners to prominence in critical care nursing. Nowhere is the management and processing of health-related information more important than in the care of the critically ill patient. Fast-paced environments, split-second decision making, wireless communications, monitoring systems run with computerized backbones, and computerized ordering and documentation, all things unimaginable just a decade ago, are now fundamental to nursing practice. Each requires a baseline understanding of informatics for true mastery. The domain of nursing informatics continues to grow as nursing incorporates expanded roles and new technology into practice. Education for nurse informaticians includes preparation from the baccalaureate level through the doctorate level and national board certification. Areas of practice are expansive, including hospitals, industry, education, policy-making, research, administration, and international settings. Although informaticians work with computers, computing technology is not the heart of the domain. Computers are simply tools that are used. Examples of informatics tools include handheld devices, point-of-care documentation, computerized provider order entry, and bar code medication administration. Nursing informatics plays an essential role in the future directions of healthcare by defining the relationship between nurses and information technology as well as the knowledge that can be gained when these domains work together. PMID- 12909796 TI - Nursing informatics, outcomes, and quality improvement. AB - Nursing informatics actively supports nursing by providing standard language systems, databases, decision support, readily accessible research results, and technology assessments. Through normalized datasets spanning an entire enterprise or other large demographic, nursing informatics tools support improvement of healthcare by answering questions about patient outcomes and quality improvement on an enterprise scale, and by providing documentation for business process definition, business process engineering, and strategic planning. Nursing informatics tools provide a way for advanced practice nurses to examine their practice and the effect of their actions on patient outcomes. Analysis of patient outcomes may lead to initiatives for quality improvement. Supported by nursing informatics tools, successful advance practice nurses leverage their quality improvement initiatives against the enterprise strategic plan to gain leadership support and resources. PMID- 12909797 TI - Clinical decision support in critical care nursing. AB - A clinical decision support system (CDSS) is a computerized application that helps clinicians detect and prevent untoward clinical events such as drug interactions, errors of omission, and trends in symptomatology. A CDSS in healthcare usually is built around an alerting system based on rules of logic. The alerting system of a CDSS can notify clinicians immediately on clinical data entry, or it can generate alerts over time after relating data from multiple sources. A CDSS for nurses and patients offers immediate benefits for nurses and patients by detecting potential drug-laboratory and drug-drug combinations and impending pharmacologic complications, monitoring microbiology results, and helping nurses relate symptoms to pharmacology and medication side effects. Other benefits include savings in time and money and reductions in morbidity and mortality. A CDSS presents an opportunity for nursing informatics and critical care nursing to collaborate for the benefit of the patient and the profession. PMID- 12909798 TI - Privacy and confidentiality: the Health Insurance Portability and Accountability Act in critical care nursing. AB - Nurses are responsible to protect the confidentiality and security of patients' health information. In the critical care setting, these privacy and confidentiality issues may be even more poignant. If able to carry on with their normal lives after discharge, many of the patients that nurses treat will have some sequelae from their illnesses that could affect their careers, finances, and personal lives. This article reviews the current literature, presents a discussion of confidentiality and security as it applies to uniquely identifiable health information, and offers some "best practices" that can be used in daily practice. Furthermore, the author discusses the Health Insurance Portability and Accountability Act of 1996 and details some reasons why the act is not fully implemented a full 6 years after it was signed into law. PMID- 12909799 TI - Human factors: imperative concepts for information systems in critical care. AB - This article provides an overview of human factors, ergonomics, human-computer interaction, and usability concepts as they relate to critical care settings. The use of these concepts can improve patient safety and the productivity of nurses, especially as they relate to the use of information systems. A framework for human-computer interaction is presented. Examples illustrate how the suite of human factors concepts is used to create intuitive, effective data presentations for use in critical care including an intensive care unit (ICU) summary display, an alternative graphic display, and modeling information sources of decision making in a neonatal ICU. The importance of integrating these concepts into advanced practice nurses' environments is made apparent. PMID- 12909800 TI - Informatics competencies for nurse practitioners. AB - Informatics knowledge and skills are essential if clinicians are to master the large volume of information generated in healthcare today. Thus, it is vital that informatics competencies be defined for nursing and incorporated into both curricula and practice. Staggers, Gassert, and Curran have defined informatics competencies for four general levels of nursing practice. However, informatics competencies by role (eg, those specific for advanced practice nursing) have not been defined and validated. This article presents an initial proposed list of informatics competencies essential for nurse practitioner education and practice. To this list, derived from the work of Staggers et al., 1 has been added informatics competencies related to evidence-based practice. Two nurse informaticists and six nurse practitioners, who are program directors, were involved in the development of the proposed competencies. The next step will be to validate these competencies via research. PMID- 12909801 TI - Acute care nurse practitioner education: enhancing performance through the use of clinical simulation. AB - Full-body patient simulators have been used for a number of years to educate nurse anesthetists and anesthesiologists. These lifelike mannequins operate from a sophisticated computerized system with the ability to generate multiple physiologic events and respond to numerous pharmacologic stimuli. The authors recently integrated the use of the patient simulator into the curriculum to educate their acute care nurse practitioner students. The learning process was divided into three steps: the presimulation experience, the simulation experience, and the postsimulation experience. These steps are described as well as important principles that need to be integrated into each phase of the process. A case scenario on respiratory failure provides an example of the simulation experience. In addition, the advantages and disadvantages of this teaching method, as identified by faculty and students, are discussed. PMID- 12909802 TI - Handheld technologies in a clinical setting: state of the technology and resources. AB - Handheld or palm-based computing technology, commonly known as personal digital assistants (PDAs), are having a tremendous impact in many personal, educational, and business settings. The potential is particularly compelling for healthcare, specifically in the clinical setting. By exploring the development of the technology, applications, and products, as well as issues regarding its use, a better understanding of this technology can be gained. The rapid acceptance and ongoing swift development of handheld computing technology is leading to an evolution in the way clinical data and information move to and from the bedside and into the patient record. This article focuses on three main areas related to PDAs: the development of the palm technology, its potential in clinical settings, and available specific resources. PMID- 12909803 TI - Use of personal digital assistants with acute care nurse practitioner students. AB - Personal digital assistants (PDAs) are attaining increased functionality by acute care nurse practitioners (ACNPs). Supplemented by recently developed medical software, these devices assist nurse practitioners in having information available at the point of care. This article reviews the introductory use of PDAs throughout ACNP graduate training with an emphasis on clinical and classroom application. PMID- 12909804 TI - Leveraging palm technology in the advanced practice nursing environment. AB - The use of personal digital assistants (PDAs) in healthcare has expanded exponentially in the past several years. In addition to common feature functions such as contact lists, calculators, calendars, and expense logs, current PDAs boast a wide variety of practical healthcare-related applications such pharmacologic databases, infectious disease programs, medication calculators, and patient scheduling and billing applications. This article examines PDAs in general and the Palm series of handheld devices in particular for use in the advanced practice setting. These devices have several implications for advanced practice nursing including support of both differential diagnosis and diagnostic reasoning, reduction of medication errors, and development of effective treatment protocols. Personal digital assistant technology will inevitably become part and parcel of advanced practice nursing. The rapid, almost daily, changes in the healthcare environment require immediate access to the myriad resources and databases used by advanced practice nurses. Personal digital assistant technology provides such access. PMID- 12909805 TI - The emerging role of electronic diaries in the management of diabetes mellitus. AB - Diabetes mellitus affects an estimated 17 million people in America. A cornerstone to the treatment of this population is appropriate glycemic control, which has been associated with better patient outcomes and cost savings from lack of diabetes complications needing treatment. The patient diary serves as the primary means used by patients and care providers to monitor and ensure glycemic control among this population and as a key component to effective diabetes management. Diabetic patients are encouraged to track continually, in a systematic way, blood glucose levels, dietary intake, activity level, and insulin dose if applicable for use in guiding appropriate care decisions. Traditionally, the recording of such patient experience data has occurred in paper-based diaries. More recently, with the advent of personal digital assistant (PDA) technology, electronic diaries are demonstrating promising benefits over paper based diaries. To take advantage of this technology, patients must be able to use PDA devices accurately and providers must be knowledgeable about their capabilities. Advanced practice nurses (APNs) in general, and nurse practitioners in the outpatient setting more specifically play a key role in assessing the appropriateness of PDA technology for individual patients, educating patients in various software applications, and ensuring the documentation and trending of data. This article provides an overview of traditional diabetes management record keeping techniques followed by a review of current PDA technologies and resources that aim to improve the state of diabetic documentation by patients. PMID- 12909806 TI - Use of personal digital assistants by adolescents with severe asthma: can they enhance patient outcomes? AB - Despite the emphasis placed on asthma education, asthma mortality is on the increase, with rates of disability higher in adolescents. Technology provides unprecedented opportunities for developing means to manage, control, and prevent acute episodes in chronic illnesses. This article describes the use of handheld wireless computer devices (eg, personal digital assistant or pocket personal computer) to prevent emergency department visits and to improve outcomes for patients with severe persistent asthma. The uses of personal digital assistants and challenges encountered in their use are described. Recommendations regarding nurses' contribution in the design, implementation, and evaluation of the program patient outcomes are provided. PMID- 12909807 TI - Genetics of common obesity and type 2 diabetes: please forget diseases and study pathogenic traits. PMID- 12909808 TI - Reverse cholesterol transport, high density lipoproteins and HDL cholesterol: recent data. AB - Unlike LDL cholesterol, which is a major cardiovascular risk factor, HDL cholesterol plays an important anti-atherogenic role through reverse cholesterol transport from peripheral cells to the liver. Some recent biochemical and epidemiological data shed light on this key function. In the hereditary Tangier disease with disseminated lipid storage, the main biochemical feature is a dramatically low level of HDL cholesterol. Different mutations in the ATP-binding cassette transporter A1 (ABCA1) gene have been recently described, which interfere with cellular cholesterol efflux. This results in low HDL plasma level, and defective reverse cholesterol transport to the liver. Moreover, selective hepatic uptake of HDL cholesteryl esters by SR-B1, a class B scavenger receptor, also plays a key role. In the follow-up of the PROCAM Study, the relative risk of coronary events is high in a cluster of patients with increased total cholesterol/HDL-cholesterol ratio. In the prospective secondary prevention VA-HIT study, the relative risk of coronary events in patients with low HDL cholesterol levels is decreased of 22% with a treatment by gemfibrozil. If the present available range of drugs targeted at increasing HDL cholesterol levels is rather narrow, future therapies will be encouraging, especially with agonists of PPARs. PMID- 12909809 TI - Sulfonylureas and cardiovascular effects: from experimental data to clinical use. Available data in humans and clinical applications. AB - OBJECTIVES: 33 years after the UGDP study, the question of deleterious effects of the sulfoylurea (SU) is still raised. We have made a systematic review of the literature from experimental studies to clinical and epidemiological studies. RESULTS: The main molecule studied is glibenclamide (GB). In vitro and in animal studies, GB is both deleterious for ischemic preconditionning (IPC) and protective for arrhythmia during acute ischemia. Glimepiride (GM) and gliclazide (GCZ) do not seem to have effect on IPC. These effects have been few studied in diabetic animals. In human, according to the investigations used, the GB seems nil or suppressing for IPC, it seems elsewhere decreases ventricular arrhythmias during periods of acute ischemia. It is possible that these two actions account for the non-appearance of concordant deleterious effects between short and long term studies. With regards to other drugs, only the GM has been specifically studied in human and appears to be nil on IPC. The only prospective clinical study available, although not having for objective to answer to this question, is the UKPDS study. This trial demonstrates the absence of deleterious cardiac effects of GB compared to chlorpropamide and particularly compared to insulin. CONCLUSION: In conclusion, in experimental studies the cardiac effects of SU differ: both deleterious and protective for GB, nil for GM and GCZ on IPC. In all cases the clinical consequences seems to be nil. PMID- 12909810 TI - The metabolic syndrome in smokers. The D.E.S.I.R. study. AB - OBJECTIVES: To test the hypothesis that smokers have a higher frequency of the metabolic syndrome, and a syndrome with different characteristics than non smokers. MATERIAL AND METHODS: The 2,569 men and 2,637 women included in the D.E.S.I.R. study, aged 30-64 years, volunteers from ten French Social Security Health Examination Centres, were studied. The World Health Organization definition of the metabolic syndrome was adapted for this analysis, and insulin resistance was defined by a concentration above the 75(th) centile of the sex specific distribution of fasting insulin. RESULTS: 28.1% of men and 15.1% of women were current smokers. The metabolic syndrome was twice as frequent in men than in women (22.8% vs 11.0%). Smokers and never-smokers were compared in age stratified samples. In men, the frequencies of hyperglycaemia or hypoglycaemic treatment, dyslipidaemia, microalbuminuria, and central obesity were significantly higher in smokers and 22.5% of smokers and 15.3% of non-smokers had this syndrome (p=0.001). In women, only dyslipidaemia was more frequent in smokers; in contrast smokers had lower insulin concentrations and hence a lower frequency of hyperinsulinemia. The frequency of the syndrome did not differ between women who smoked and those who did not (6.3% vs 6.0%). CONCLUSIONS: In men, the metabolic syndrome was more frequent in smokers than in non-smokers; in contrast, there was no difference for women. For both sexes, syndrome abnormalities tended to be more frequent in smokers than in non-smokers. PMID- 12909811 TI - Time trend in height, weight, and obesity prevalence in school children from Northern France, 1992-2000. AB - OBJECTIVE: The aim of this study was to evaluate the changes in height, weight and in the prevalence of overweight and obesity between 1992 and 2000 in children living in two towns of northern France. METHODS: Two cross-sectional studies were performed in every pre-school (last section) and primary schools of the two towns in 1992 (383 girls and 421 boys enrolled) and 2000 (296 girls and 305 boys). Children were 5 to 12 year old in both studies. MEASUREMENTS: Body height and weight were measured, and BMI was calculated (weight/height(2)). Prevalence of overweight and obesity was determined according to the gender- and age- specific cut-offs of the new international reference (IOTF). We also used the 90(th) and the 97(th) percentiles of the French reference gender- and age-specific BMI curves to define two grades of overweight. RESULTS: After adjustment for age, boys were on average 1.5 cm taller in 2000 than in 1992 (p<0.001), and the same trend was observed in girls (+ 0.9 cm, p<0.075). Height-adjusted or age-adjusted weight and BMI were significantly higher in 2000 than in 1992. In girls, obesity defined by IOTF criteria increased from 1.6 to 4.4% (p<0.03) and overweight from 14.1 to 18.6% (p<0.11). In boys, the change in prevalences was significant only when the less stringent criteria (i.e. the 90(th) percentile of French references) was used (13.8% in 1992 vs 20% in 2000, p=0.03). CONCLUSION: Over an 8 years period, there was an increase in height and BMI in both boys and girls. These results show that the increase in the prevalence of obesity is accompanied by a global trend of accelerated growth. PMID- 12909812 TI - Management and short-term outcome of diabetic patients hospitalized for acute myocardial infarction: results of a nationwide French survey. AB - OBJECTIVES: To compare management and short-term outcome of diabetic and non diabetic patients hospitalized for acute myocardial infarction. METHODS: This was a prospective epidemiological survey. All patients admitted in coronary care units in France in November 2000 for confirmed acute myocardial infarction were eligible to enter the study. RESULTS: Of the 2320 patients recruited from 369 centers, 487 were diabetic (21%). Compared to non-diabetic patients, diabetic patients were 5 years older, more often female, obese and hypertensive; they had more often a history of cardiovascular disease; they had a lower ejection fraction and worse Killip class. Reperfusion therapy was less frequent among diabetic patients (39% versus 51%; p=0.0001), as was the use of beta-blockers (61% versus 72%; p=0.0001), aspirin (83% versus 89%; p=0.0001) and statins (52% versus 60%; p=0.001) during hospitalization. Conversely, the use of ACE inhibitors was more frequent (54% versus 44%; p=0.0001). 58% of diabetic patients received insulin during hospitalization. Twenty-eight-day mortality was 13.1% in diabetic patients and 7.0% in non-diabetic patients (risk ratio: 1.87; p=0.001). Diabetes remained associated with increased mortality after adjustment for relevant risk factors including age and ejection fraction (risk ratio: 1.51; p=0.07). In patients treated with antidiabetic drugs (chiefly sulfonylureas) before admission, 28-day mortality was 10.4% compared with 19.9% in diabetic patients on diet alone or untreated (p=0.005). CONCLUSION: Despite higher cardiovascular risk and worse prognosis, in-hospital management of diabetic patients with acute myocardial infarction remains sub-optimal. Patients previously treated with antidiabetic drugs including sulfonylureas had a better prognosis than untreated diabetic patients. PMID- 12909813 TI - Prospective audit of diabetes care and outcomes in a group of geriatric French care homes. AB - BACKGROUND: In geriatric institutions, diabetes prevalence varies from 10 to 20%. However, little is known about patterns of care and their outcomes. To assess both, an 18-month prospective audit was conducted in 240-bed nursing and 80-bed residential care homes. METHODS: In diabetic subjects, items of care were compared to the corresponding French guideline end-points. Dependency in activities for daily living (ADL) was evaluated at inclusion and at the end of the survey for survivors. Diabetic residents were compared to others for age, sex, mortality and one-year change of iso-resource group ranging (IRG), a need-of care scale. RESULTS: The 73 diabetic patients among 494 residents (14.8%) were not different from the others for age and sex (respectively age: 76.0 y +/- 7.9 compared to 78.2 y +/- 9.4; M/F sex ratio: 0.59 compared to 0.31). Oral hypoglycemic agents were prescribed in 29 (39.7%) and insulin in 26 (35.6%). Control of diabetes with HbA(1C) was in keeping with guidelines in 23.3%, HbA(1C) was never performed in 26%, and > 8% in 20.5%. Rates of items of guidelines that were never addressed ranged from 2 to 80%. During the stay, ADL dependency worsened in diabetic patients, but their one-year change in GIR range was not different from that in the others. Mortality was 30.1% compared to 37.1% in the others (NS). This mortality rate and the evolution of functional dependency were not influenced by the quality of diabetes control. CONCLUSION: In these disabled elderly patients, diabetes management needs to be improved. However, mortality and the evolution of functional dependency were similar in diabetic people than in others. Interventional studies for controlling with reference to geriatric evaluation, i.e. function, nutrition and mental health, are needed in order to establish specific guidelines based on benefit-burden analysis. PMID- 12909814 TI - Screening diabetic patients at risk for foot ulceration. A multi-centre hospital based study in France. AB - BACKGROUND: To determine the prevalence of risk factors for diabetic foot ulceration in diabetic patients free of active pedal ulceration in a hospital setting. METHODS: In sixteen French diabetology centres, a survey was conducted on a given day in all diabetic people attending the units, both as in- or out patients. RESULTS: 664 patients were evaluated: 105 had an active foot ulcer and were excluded from the analysis as were four other patients due to lack of reliable data. From the 555 assessable patients, 40 (7.2%) had a history of foot ulcer or lower-limb amputation. Sensory neuropathy with loss of protective sensation, as measured by the 5.07 (10 g) Semmes-Weinstein monofilament testing, was present in 27.1% of patients, whereas 17% had a peripheral arterial disease mainly based on the clinical examination. On addition, foot deformities were found in 117 patients (21.1%). According to the classification system of the International Working Group on the Diabetic Foot, 72.8% of patients were at low risk for pedal ulceration (grade 0) and 17,5% were in the higher-risk groups (grade 2 & 3). If patients with isolated peripheral arterial disease were considered as a separate risk group (as was those with isolated neuropathy), percentage of low-risk patients decreased to 65.6%. There was a clear trend between the increasing severity of the staging and age, duration of diabetes, prevalence of nephropathy and retinopathy. CONCLUSIONS: Prevalence of risk factors for foot ulceration is rather high in a hospital-based diabetic population, emphasising the need for implementing screening and preventive strategies to decrease the burden of diabetic foot problems and to improve the quality of life for people with diabetes. PMID- 12909815 TI - An economic evaluation of the cost of diabetic foot ulcers: results of a retrospective study on 239 patients. AB - OBJECTIVE: To evaluate the cost of foot ulcers in diabetic patients. METHODS: Retrospective pharmacoeconomic study using direct and indirect costs (sick leave days) from the perspective of French social security system. RESULTS: 239 patients were included in the study by 80 physicians who treat diabetic patients suffering from foot ulcers. Initially identified by telephone survey, these physicians were primarily endocrinologists/diabetologists, general practitioners and surgeons. Average monthly costs in the treatment of foot ulcers were 697 euro; for outpatient care, 1556.20 euro; for hospital care (day treatment and short stays), and 34.76 euro; for sick leaves. When hospitalization was required, it represented approximately 70% of the average cost for foot ulcers. The portion of outpatient costs was principally generated by medical and paramedical treatments, and interventions carried out by healthcare personnel. On the other hand, medication only represented 10% of total costs. The initial severity of the pathology was a determinant clinical factor of high healthcare costs. In addition, the more recent the lesion was, the higher the cost of treatment. Amputation and follow-up by specialists were correlated to high costs as well, a logical result of these clinical factors. CONCLUSION: This analysis is the first to evaluate the cost of treating foot ulcers in such a large population of diabetic patients. The economic outcomes should help direct public authorities in their choices, particularly as regards the interest of treating these diabetes related complications as early as possible. PMID- 12909816 TI - Kinetics of plasma and erythrocyte metformin after acute administration in healthy subjects. AB - OBJECTIVES: Although the existence of a deep compartment for metformin has long been hypothesized, there is still little direct information concerning metformin distribution in individual tissues in man. The only available study involves chronic metformin therapy. In that study, the measurement of metformin in erythrocytes provided a reliable indicator of metformin distribution and of potential accumulation. To determine the kinetics of metformin in plasma and in erythrocytes after acute oral administration, we performed the present study in healthy subjects after a single oral dose of metformin and compared the pharmacokinetics parameters in erythrocytes to those in plasma. METHODS: Six nondiabetic participants took the study dose of 850 mg metformin at 8: 00 AM after a non-standardized breakfast (i.e., as recommended in clinical practice). Blood samples were collected for metformin measurement in plasma and in erythrocytes at 0, 1, 2, 3, 4, 6, 9, 24, 33, 48, 57, and 72 h. RESULTS: Maximum metformin concentration was attained at 3.0 +/- 0.3 h in plasma and 4.7 +/- 0.5 h in erythrocytes. This difference was not significant. Metformin concentrations peaked at a maximum almost 6 times higher in plasma than in erythrocytes (1.7 +/- 0.1 and 0.3 +/- 0.0 mg/l, respectively). However, because the elimination half life of metformin was much longer in erythrocytes (23.4 +/- 1.9 h vs. 2.7 +/- 1.2 h), there was no difference in area under the curve between plasma and erythrocytes. The distribution volume (plasma) was calculated to be 146 +/- 11 l. Plasma and erythrocytes concentration-time curves showed that metformin was not detectable in plasma 24 hours after the oral administration, while it remained detectable in erythrocytes up to 48 hours. Metformin concentrations crossed approximately 13 hours after having reached their maximum values in plasma, approximately 16 h after metformin intake. CONCLUSION: Having demonstrated the rapid elimination of metformin from plasma and its slow disappearance from erythrocytes, the presents results should contribute to adjustment of metformin dosage to renal function, assessment of drug compliance, and retrospective analysis (when blood samples are drawn with delay) of the link between metformin and development of lactic acidosis. Most importantly, the present findings should help to ascertain the optimal dosage of metformin, particularly in elderly patients. PMID- 12909817 TI - What kind of simple fasting index should be used to estimate insulin sensitivity in humans? AB - The hyperinsulinemic euglycemic glucose clamp method is the gold standard for measuring insulin resistance. However it is complex, and simple indexes have been developed. Some of them are based on formulae that calculate the product or the addition of fasting plasma insulin and glucose values whereas others are based on their ratios. We calculated several simple indexes of insulin resistance and compared them to hyperinsulinemic euglycemic clamp data in 111 subjects with a wide range of insulin resistance. We showed that indexes using insulin and glucose ratios in their formulae are poorly correlated with clamp measurements and give false evaluations, particularly in glucose-intolerant and type 2 diabetic subjects. Thus, whatever the glucose profile of study subjects, we suggest the use of a simple index based on the product or the addition of fasting plasma insulin and glucose values instead of their ratios to obtain insulin resistance evaluations close to the hyperinsulinemic euglycemic clamp technique. PMID- 12909819 TI - Atypical neuroleptics and diabetes. AB - Since a few years, an increasing number of cases of atypical neuroleptic associated diabetes are reported in the literature. But few are dedicated to diabetologists. We report here two cases of patients with a severe deterioration of preexisting diabetes, with clozapine for the first case report and olanzapine for the second one. We also make a literature review and discuss the possible mechanisms involved in this atypical neuroleptic-associated diabetes. We recommend a thigh follow-up of weight, glycemic and lipidic parameters during psychotic patients treatment with atypical neuroleptics, in order to prevent or rapidly treat the metabolic complications described in the literature. PMID- 12909818 TI - Fish oil prevents the adrenal activation elicited by mental stress in healthy men. AB - OBJECTIVES: A diet rich in n-3 fatty acids (fish oils) is associated with reduced risks of cardiovascular and metabolic diseases, but the mechanisms remain incompletely understood. Sympathoadrenal activation is postulated to be involved in the pathogenesis of these diseases, and may be inhibited by n-3 fatty acids. We therefore evaluated the effects of a diet supplemented with n-3 fatty acids on the stimulation of the sympathetic nervous system and of stress hormones elicited by a mental stress. METHODS: Seven human volunteers were studied on two occasions, before and after 3 weeks of supplementation with 7.2 g/day fish oil. On each occasion, the concentrations of plasma cortisol, and catecholamines, energy expenditure (indirect calorimetry), and adipose tissue lipolysis (plasma non esterified fatty acid concentrations) were monitored in basal conditions followed by a 30 min mental stress (mental arithmetics and Stroop's test) and a 30 min recovery period. RESULTS: In control conditions, mental stress significantly increased heart rate, mean blood pressure, and energy expenditure. It increased plasma epinephrine from 60.9 +/- 6.2 to 89.3 +/- 16.1 pg/ml (p<0.05), plasma cortisol from 291 +/- 32 to 372 +/- 37 micromol/l (p<0.05) and plasma non esterified fatty acids from 409 +/- 113 to 544 +/- 89 micromol/l (p<0.05). After 3 weeks of a diet supplemented with n-3 fatty acids, the stimulation by mental stress of plasma epinephrine, cortisol, energy expenditure, and plasma non esterified fatty acids concentrations, were all significantly blunted. CONCLUSION: Supplementation with n-3 fatty acids inhibits the adrenal activation elicited by a mental stress, presumably through effects exerted at the level of the central nervous system. PMID- 12909820 TI - [Diabetic retinopathy in children and adolescents]. AB - Diabetic retinopathy rarely occurs before puberty and is never proliferative in prepubescent children. On the opposite, puberty and adolescence are high-risk periods for diabetic retinopathy progression, and call for strict ophthalmologic monitoring. The period between 16 and 18 years of age is particularly critical. Progression towards florid diabetic retinopathy is to be especially feared and should be prevented in the course of adolescence, as this form can be severe and can lead to blindness. Risk factors are probably many, including diabetes duration, difficulties in achieving glycemic control due to increase in insulin requirements, low compliance to treatment, and hormonal changes related to puberty (abnormalities of the growth hormone (GH)/insulin-like growth factor-I (IGF-1) axis). Systematical diabetic retinopathy screening should be performed in adolescents, notably by non-mydriatic fundus photographs. Furthermore, the anticipation of the switch from pediatric to adult structures, together with the careful information and education of parents and children may improve visual prognosis of young diabetic patients, whose life expectancy is high. PMID- 12909821 TI - [French diabetologists' standpoint on the prevention of type 2 diabetes. A survey carried out during the ALFEDIAM Convention Bordeaux 2003]. AB - An opinion poll was carried out during the ALFEDIAM Congress Bordeaux 2003. One hundred and thirty-seven participants (mean age 43.6 +/- 8.3 years/sex Ratio approximately 1) among whom 22.6% run private practices, 51.8% work in hospitals and 21.3% are both private and hospital practitioners, have been questioned about their conception of the prevention of type 2 diabetes. Prediabetes is an acknowledged entity for 61% of the people surveyed. Two thirds use as a diagnostical criterion, moderate fasting hyperglycemia and/or a impaired glucose tolerance. Oral glucose tolerance test (OGTT) is still commonly practised among 51.9% but that is done sparingly only to confirm the diagnosis of diabetes in presence either of several risk factors or of a moderate fasting hyperglycemia. According to 70% of the answers, the detection of diabetes must be repeated every year among at risk subjects aged over 45. The metabolic syndrome is defined according to diverse criteria. The right definition of ATP III is given only in 5% of the cases. As regards the treatment, the combined requirements of physical activity and dietary rules are approved by 97% of the answers. The majority of the persons questioned in the survey consider that a slight loss of weight (less than 5% of the initial weight) is sufficient in a high risk risk individual.On the other hand, opinions are divided as regards the use of drugs at the pre diabetes stage. Metformin is the only one that is accepted by more than 50% with a rate of 58.4% of positive answers, acarbose and orlistat rating respectively 37.2% and 35%. However a great majority (83.6%) are in favour of the reimbursement of antidiabetic drugs in this indication, for high risk individuals, provided a study has clearly demonstrated the efficiency of the molecule concerned. PMID- 12909822 TI - [Diagnosis of adult chronic kidney failure]. PMID- 12909823 TI - [Tuberculosis and non tuberculous mycobacterial diseases]. PMID- 12909824 TI - [Value of PCR amplification from formalin-fixed paraffin-embedded tissues in the diagnosis of Mycobacterium tuberculosis infection]. AB - Mycobacterium tuberculosis infection remains a major health problem throughout the world. In France, tuberculosis is endemic, particularly in the Paris area (Ile-de-France) and in the south (Provence Alpes cote d'Azur) where immigration is greater than in other countries in northern Europe. Culture is the gold standard for diagnosis of tuberculosis and is the only method enabling a study of strain sensitivity to treatment. Histology contributes to diagnosis in most cases by revealing typical necrotizing granulomatous lesions. The diagnosis is then confirmed by the detection of acid-fast bacilli with Ziehl-Neelsen staining. However, the Ziehl-Neelsen stain is not sensitive and does not allow identification of different species. The polymerase chain reaction (PCR) DNA amplification method has been used to detect M. tuberculosis in formalin-fixed paraffin-embedded tissues. The aim of the present study was to investigate the value of this method for the diagnosis of M. tuberculosis infection. The results obtained with PCR assay were compared to those obtained with histological and microbiological methods (direct examination and culture). Sixty-three specimens (mainly lymph node and lung specimens) exhibiting a positive culture for M. tuberculosis were analyzed. Tuberculosis granulomas were noted in 32/63 cases, tuberculoid granulomas in 18/63, pyoepitheloid granuloms in 10/63, and non specific inflammation in 3/63. Ziehl-Neelsen staining was positive in 11/63 cases. PCR assay on tissue sections was positive for M. tuberculosis in 58/63 cases. Controls of the PCR method (granulomas due to other mycobacterial species, foreign body granulomas, sarcoidosis granulomas) were all negative. This study shows that PCR from deparaffinized sections, 1) greatly increases the sensitivity of diagnosis of tuberculosis, 2) enables the diagnosis of M. tuberculosis infection. However, although this method reduces the time to diagnosis, culture remains the gold standard for identification of the mycobacterium and for determining the sensitivity of the isolated strain to treatment. PMID- 12909825 TI - [Non tuberculous mycobacterial diseases]. AB - Mycobacteria species other than members of Mycobacterium tuberculosis complex are called non tuberculous mycobacteria (NTM) or "atypical" mycobacteria. To date, about 80 mycobacterial species have been described. They are usually opportunistic pathogens with variable degrees of virulence. Tuberculosis is the commonest mycobacterial disease in the world, followed by leprosy and Buruli ulcer. Before the AIDS epidemic, NTM diseases were confined to the lungs (M. kansasii, M. intracellulare and M. avium), the skin (M. marinum) or cervical lymph nodes (M. scrofulaceum). The outbreak of AIDS epidemic has dramatically changed the epidemiology of NTM diseases. Between 25 to 50% of AIDS patients in Europe and USA are infected with NTM. NTM infections are usually disseminated in patients with profound immunodeficiency. The inflammatory response and the prognosis of NTM diseases depend on the immunological status and the NTM species. Diagnosis may be difficult, especially in AIDS patients in whom numerous diseases are often associated. Diagnostic criteria involve clinical, radiological, microbiological and pathological findings. Identification of Mycobacterium species in cultures is the gold standard. Pathological examination has several interests: it may reveal an NTM disease, it provides a more rapid assessment of the infection than cultures, and helps to evaluate the virulence of NTM species identified by cultures. PMID- 12909826 TI - [Placental site trophoblastic tumor of the uterus after in vitro fertilization. A case report with initial vaginal metastasis]. AB - We report the first case of placental site trophoblastic tumor following in vitro fertilization. A 32-year-old woman presented with a vaginal polyp diagnosed 11 months after a successful fertilization, resulting in the delivery of a term male baby, but followed by persistent amenorrhea. The diagnosis of PSTT was done on examination of the vaginal polyp and the curettage specimen and was confirmed after hysterectomy. The patient was well 30 months later. The value of initial metastases limited to the pelvis and the role of in vitro fertilization in the pathogenesis of the tumor are discussed. PMID- 12909827 TI - [Non-differentiated carcinoma with osteoclast-like giant cells of the pancreas]. AB - We report the case of a 53-year-old man who presented a mass involving the head of the pancreas resulting in isolated jaundice. Histologically, the tumor was an undifferentiated carcinoma with osteoclast-like giant cells of the pancreas. This very rare neoplasm resembles giant cell tumor of bone. An epithelial origin is now established and this tumor has been recently considered as a variant of ductal adenocarcinoma of the pancreas in the last WHO histological classification. The diagnosis requires both morphology and immunohistochemistry. Although the prognosis of these tumors is reported to be poor, our case is unusual because of a favorable outcome without relapse after 2 years. PMID- 12909828 TI - [Neurectodermal differentiation of extraskeletal myxoid chondrosarcoma: a classical feature?]. AB - Extraskeletal myxoid chondrosarcoma (EMC) is a phenotypically and genotypically distinct entity with a protracted course. A documented case of an extraskeletal myxoid chondrosarcoma characterized by a t(9; 17) (q22; q11) translocation with a neuroendocrine and neural differentiation is reported. PMID- 12909829 TI - Histiocytoid breast carcinoma: a case report of an uncommon histologic variant of lobular carcinoma. AB - A rare case of invasive histiocytoid breast carcinoma is presented. A middle aged female patient underwent quadrantectomy for a palpable mass in her right breast. The neoplastic cells showed pronounced histiocytoid appearance and immunopositivity for cytokeratins 7 and CAM 5,2, EMA, CEA, GCDFP-15, MFG and ER/PR. We report here this uncommon histologic pattern because of the considerable diagnostic interest. Recognition of this histologic variant either in primary or metastatic locations may present some difficulties to the pathologist. Differential diagnostic problems are emphasized and the literature is reviewed. PMID- 12909830 TI - [Extraskeletal myxoid chondrosarcoma of the nasopharynx]. AB - Extraskeletal myxoid chondrosarcoma is a rare but distinct entity with special clinicopathological, immunohistochemical, cytogenetical and outcome features. This tumor developed from soft tissues. A few cases have been reported in the head and neck in the literature. We report two new cases of extraskeletal myxoid chrondrosarcoma presenting in such an unusual site: one involved the left nasal cavity of a 67-year-old man and the second the sphenoidal sinus of a 71-year-old woman. The microscopic examination revealed nests of round small cells dispersed in a myxoid stroma. The myxoid material was stained with Alcian Blue with and without hyaluronidase application whereas no PAS staining was observed. The immunohistochemical staining showed reactivity with S-100 protein and vimentin in two cases and with EMA in one case. These results allowed us to exclude other differential diagnoses: soft tissue tumors with a myxoid stroma (myxoma, myxoid liposarcoma and myxofibrosarcoma). No staining with anti-KL1 allowed us to exclude chordoma. Curative surgery was not possible. Both patients were given radiotherapy and the tumor regressed in one. PMID- 12909831 TI - [Unusual parosteal osteochondromatous proliferation or Nora's tumor. A clinicopathological analysis of 4 cases]. AB - Nora's lesion, also known as bizarre parosteal osteochondromatous proliferation (B.P.O.P.), involves mostly the small tubular bones of the hands and feet. Histologically, it is characterized by a proliferation of chondroid, bony and fibrous tissues, sometimes with high cellular density, bizarre chondrocytes but is devoid of cellular atypia and necrosis. Distinct blue color is noted at the interface of bone and cartilage. The most important lesions that present differential diagnostic problems are chondrosarcoma, parosteal osteosarcoma and florid reactive periostitis. The lesion is benign but may recur locally in as many as 55%. The clinical and pathological findings of four cases of Nora's lesion are presented. PMID- 12909832 TI - [Tuberculosis: a rare and misleading etiology of tongue's ulcer]. AB - We report the case of a 43 year-old man, smoker, who used to live in Africa, consulting for a chronic ulcer of the mobile tongue. An initial biopsy did not show any carcinoma. A second biopsy highlighted an inflammation with numerous tuberculoid granulomas. However, the Ziehl-Neelsen stain was negative. Histoplasmosis of the tongue was then suspected as some round structures looking like yeasts and stained by the Gomori Grocott method were seen within the cytoplasm of giant cells. However, immunohistochemistry using anti-Histoplasma antibodies was negative. Polymerase chain reaction (PCR) assay performed on deparaffinized sections allowed the diagnosis of infection by Mycobacterium tuberculosis. A third biopsy confirmed the diagnosis of tuberculosis by showing some exceptional acid-fast bacilli. Culture was negative. Tuberculosis of the tongue is a very rare condition with different differential diagnosis including carcinoma in smoker population or histoplasmosis in endemic area. PMID- 12909833 TI - [2002 Standards, Options and Recommendations: good practice for the management and shipment of histological and cytopathological cancer specimens]. AB - CONTEXT: The Standards, Options and Recommendations (SOR) collaborative project was initiated in 1993 by the Federation of the French Cancer Centres (FNCLCC), with the 20 French Regional Cancer Centres, several French public university and general hospitals, as well as private clinics and medical speciality societies. Its main objective is the development of serviceable clinical practice guidelines in order to improve the quality of health care and the outcome of cancer patients. The methodology is based on a literature review, followed by critical appraisal by a multidisciplinary group of experts. Draft guidelines are produced, then validated by specialists in cancer care delivery. OBJECTIVES: Produce clinical practice guidelines for the management and shipment of histological and cytopathological cancer specimens using the methodology developed by the Standards, Options and Recommendations project. METHODS: The FNCLCC designated the group of experts. Available data were collected by a search of Medline and lists selected by experts in the group. A first draft of the guidelines was written, then validated by independent reviewers. RESULTS: The main recommendations are: 1/ High-quality transmission of information between professionals is essential to the management of cancer specimens in order to assure high-quality diagnosis and evaluation of prognostic factors; 2/ Written procedures concerning sample shipment, handling, storage, registration, tracking and fixation exist; these procedures, as well as the necessary shipping material, will be sent to all clinical services involved; 3/ When possible, fresh, unfractionated, oriented surgical samples will be submitted to the same histological and cytopathological laboratory; 4/ Samples collected for extemporaneous examination, freezing or cell culture must be shipped immediately under appropriate storage conditions; 5/ Once frozen, samples can be stored in a deep freezer at temperatures of -80 degrees C or below, or kept in liquid nitrogen; 6/ Fixing tissues shortly after sample collection is essential to prevent cell lysis; 7/ Computerised systems will be used to assure correct specimen registration and tracking in histological and cytopathological laboratories. PMID- 12909834 TI - [A third testis!]. PMID- 12909835 TI - [Unusual cells in the mesentery]. PMID- 12909836 TI - [An uncommon and misleading neoplasm to know!]. PMID- 12909837 TI - [A thyroid neoplasm]. PMID- 12910028 TI - [Thinking of cocaine]. PMID- 12910030 TI - [Analysis of drug consumption among ambulatory patients in a drug agency of Northern France (Lens)]. AB - The aim of this study was to describe and analyze the patterns of illicit psychoactive drug consumption, and the utilization of buprenorphine (BHD) in opiate-dependent patients interviewed in the drug agency of Lens. In our sample, multiple drug use was a massive phenomenon. Misuse of BHD was related to young age, multiple drug injection, gender (male) and risky injecting behavior. Multiple drug use was less frequent among patients with a BHD prescription. It is urgent that health professionals and decision-makers be aware of the heterogeneity of drug consumption habits and their impact on the utilization of drug maintenance treatment. Research is needed to understand the history and motivations for misuse and persistent injecting among some drug users. A national conference of consensus on how to implement and maintain treatments is a necessary step forward in caring for opiate drug addicts. PMID- 12910029 TI - [High-dose buprenorphine prescription in southern France from 1999 to 2001]. AB - Several reports have focused on prescription of high-dose buprenorphine using data electronically transmitted to the French health reimbursement system. This study deals with high-dose buprenorphine prescriptions in the Bouches-du-Rhone region between 1999 and 2001. We determined the number of maintained patients followed by practitioners (either short- or long-term care), the doses administered, and the associated prescription of psychotropic treatments. The number of general practitioners involved in 84% of the prescriptions of high-dose buprenorphine grew with a high turnover (20%). More than 25% of the general practitioners in the region prescribed this drug, but only 38% of them were involved in long-term follow-up of more than one maintained patient. The number of maintained patients and the mean daily treatment dose delivered (13.4 mg/d during 2001) increased. The associated prescriptions of benzodiazepines concerned 47% of the patients during 2001 (on the rise), but the associated prescription of flunitrazepam tended to decrease (29% of the subjects during 2001 versus 32% during 2000). This study shows a tendency for deviant behaviors to increase, constituting a public health concern. PMID- 12910031 TI - [Sporting activities and psychoactive substance use. Data abstracted from the French part of the European School Survey on Alcohol and other Drugs (ESPAD 99)]. AB - Few studies have analyzed in the general population psychoactive substance use among athletes, especially among females. In fact, sporting activity is often promoted in prevention actions, as an alternative to addiction or alcohol, tobacco or other substance misuse. So, we propose an analysis of the ESPAD 1999 sample among students (16-18 years old), focused on the relationship between sporting activities and substance use. Boys play sport more frequently than girls (71.5% versus 49.5%) and report 8 hours and more a week 4 more times than girls (14% versus 3.5%). Sixty-eight percent of boys and 36% of girls have already participated in sport competitions, more often at a local, departmental or regional level; a minority of them (26% of boys and 20% of girls) have already participated in sport competitions at a national or international level. Sporting activity is decreasing with age among girls, students from general lycee play sport more frequently than others do (vocational lycee); the higher the father's education level, the more frequently the students play sport. Moderate sporting activity (1-8 hours a week) is a protective factor against regular smoking (OR=0.54 in boys and OR=0.60 in girls) and against regular cannabis use among boys (OR=0.64). Intensive sporting activity (>8 hours a week) is a risk factor for illicit drugs (except cannabis) use (OR=2.74) and sleeping drugs/tranquillizers (OR=1.82) only among girls. Competition level is the most important risk factor for substance misuse as well in boys (except sleeping drugs/tranquillizers) as in girls. Practical implications are: adjusting health policy concerning the beneficial effects of sporting activity, raising sports associations abilities and avoiding doping and addiction in high-level sporting activities. PMID- 12910032 TI - [High dosage buprenorphine and injection practices. A study of 303 patients]. AB - In France, by the end of 1999, a study of a naturalistic-type was led by the Louis-Harris Institute on 303 persons taking high dosage (HD) buprenorphine. This study aimed to identify factors likely to be correlated with stopping or continuing HD buprenorphine injections. We carried out a comparative study of four groups of HD buprenorphine users: "non-injectors" (n=90), "ex-injectors" (n=71), "monitored injectors" (n=69) and "un-monitored injectors" (n=72), with intra-group representativeness. The data was gathered in the context of anonymous interviews, by objective interviewers, in 20 regions. Most of the interviewees were also users or ex-users of more than one psychoactive substance. The "un monitored injector" group was younger than the "non-injector" group and not as well integrated into society. The injection frequency was lower in patients receiving medical care. Research into the reasons for stopping and cutting down on HD buprenorphine injections revealed indicators such as the impossibility of breaking the injection habit, the fact that users seek the immediate sedation effect, and associating with friends who also inject. These factors seem to be greatest during critical periods, such as depression or the absence of well being. We showed that the factors involved in cutting down included: revaluation of the way in which the medicine was taken with the doctor, and becoming aware of the problems resulting from injection through information received from the doctor and/or by experiencing problems firsthand. In all the groups, the benefits associated with taking HD buprenorphine were observed and evaluated, starting with a list of 14 items. Five main benefits were found: cutting down on or stopping heroine, taking better care of oneself, making new plans, being in better physical shape, and finally, sleeping better. The improvement was greatest in the "non-injector" and "ex-injector" groups. The work that would be necessary to establish effective medical support involves a global approach including an appropriate initial prescription, paying special attention during critical periods, and verification of the mode of use so as to avoid under-dosing, and constant evaluation of the risk of resorting to injections. The fact that there are ex-injectors and that some regular injectors have not resorted to injecting during the last month shows the benefits of medical support. PMID- 12910034 TI - Methadone therapy in Croatia. PMID- 12910033 TI - [Voluntary poisoning by ingestion of Datura stramonium. Another cause of hospitalization in youth seeking strong sensations]. AB - Natural hallucinogenic substances have been used in numerous cultures throughout the world for millenniums, providing a sacred intermediary between men and gods. They were used during initiatory, divinatory rites and played an essential role in the myths of exorcism and charm where the initiated enters a sort of trance which separates the soul from the body before "revival". It resulted that a number of plants were given names reflecting their relationship with the divinities, bestowing upon them an aura of mystery and sacredness. With the development of free information superhighways via the net, recipes using these hallucinogenic substances can now travel anywhere on the planet, offering the non initiated a means of experimenting their search for strong sensations. At the same time, the science of botany has undergone a major revolution. Patient observation of nature is no longer necessary since a well-conducted computer search can easily yield a large supply of plants offered for on-line purchase. Children and adolescents in primary and secondary schools are becoming a new target for the botanical market: young people exchange the latest recipes and Internet addresses, or more simply the objects of their new on-line purchase. This has led to an outbreak in the use of mushrooms and hallucinogenic plants where the cultural notion of initiation or sacred rites has metamorphosed into a phenomenon of communication between young people, notably during festive parties. Datura stramonium has become quite popular and the number of hospitalizations for acute datura poisoning has increased rapidly, as indicated by the statistics of the Bordeaux Poison Center (14 hospitalizations for voluntary poisonings with datura in 2002, compared with 5 in 2001). Besides the self-inflicted risk, one of the most important problems is the delay to diagnosis, simply because the primary care physician did not think of such an "exotic" cause. We report here the case of a 22-year-old young man hospitalized following ingestion Datura stramonium seeds. He presented confusions and dreamlike frenzy. The clinical course was favorable in 72 hours with symptomatic treatment, marked by amnesia of the episode. PMID- 12910035 TI - [Maintenance treatment for opiate dependence. Which should we deliver and how should we prescribe it?]. AB - Dr Olivier Jacquiez has practised in the Specialised Care Centre of Dunkerque for 3 years. From his own clinical experience related to the use of methadone and the medical literature, he gives his points of view concerning maintenance treatment delivery, and the eventual influence of the regulations promulgated on January 30th, 2002 by the French Health Authorities (DGS/DHOS 2002/57) allowing each hospital practitioner to initiate a drug maintenance treatment with methadone. PMID- 12910036 TI - [Cognitive disorders and chronic use of cannabis]. AB - Cannabis is by far the illegal substance the most widely used by youth aged 12-25 years. One out of five persons living in France has already tried it once in his lifetime. Although the psychiatric symptoms as well as the cognitive and acute or chronic behavioral effects linked to repeated use of cannabis are well documented in the literature, search for persistent cognitive effects amongst chronic users has not provided convincing evidence because of methodological biases. The issue is nevertheless most crucial, especially amongst teenagers, due to the potential risks of deteriorated academic, social and occupational performance. The longer the exposure to the effects of the substance, the greater the risk of complications. Based on data reported in the literature, we report the cognitive effects associated with chronic use of cannabis as well as its social and educational consequences. PMID- 12910037 TI - [Relationships between alexithymia, depression and interpersonal dependency in addictive subjects]. AB - In the scientific literature, the term of addiction is currently used to describe a whole range of phenomena characterized by an irresistible urge to engage in a series of behaviors carried out in a repetitive and persistent manner despite accruing adverse somatic, psychological and social consequences for the individual. It has been suggested that subjects presenting such behaviors would share specific features of personality which support the appearance or are associated with these addictive behaviors. Dimensions such as alexithymia and depression have been particularly well investigated. The aim of this study was to explore the hypothesis of a specific psychopathological model relating alexithymia and depression in different addictive disorders such as alcoholism, drug addiction or eating disorders. Alexithymic and depressive dimensions were explored and analyzed through the statistical tool of path analysis in a large clinical sample of addicted patients and controls. The results of this statistical method, which tests unidirectional causal relationships between a certain number of observed variables, showed a good adjustment between the observed data and the ideal model, and support the hypothesis that a depressive dimension can facilitate the development of dependence in vulnerable alexithymic subjects. These results can have clinical implications in the treatment of addictive disorders. PMID- 12910038 TI - [New challenges in medical oncology. From medical oncology to oncological medicine]. PMID- 12910039 TI - [Mesenteric venous thrombosis. A retrospective study of 23 cases]. AB - Mesenteric venous thrombosis is uncommon. Diagnosis is difficult and much remains to be learned about clinical aspects, etiology and treatment options. PATIENTS AND METHODS: We conducted a retrospective study over 5 years concerning patients with isolated mesenteric venous thrombosis or with extension to the portal vein. RESULTS: Twenty-three patients (12 women), mean age 55 years, were included. Abdominal pain (96%), bowel obstruction (35%), fever (26%), diarrhea (17%), gastrointestinal bleeding (13%) were the most common symptoms at presentation. In 78% of patients, at least one underlying condition was identified: thrombophilia, oral contraceptive, cancer, paroxysmal nocturnal hemoglobinuria, essential thrombocythemia, intra-abdominal processes. CT scan and ultrasound diagnostic sensitivity were 89% and 57% respectively. Outcome was favorable for the 18 patients treated with anticoagulants (associated with surgery in 4 patients). Three patients with neoplasm died early. CONCLUSION: Mesenteric venous thrombosis must be kept in mind as a possible diagnosis in patients with abdominal pain. Predisposing factors are frequently associated and should be searched for. Early diagnosis, using CT scan or ultrasound, enables immediate anticoagulation and seems to improve outcome. PMID- 12910040 TI - [Treatment of malignant non-Hodgkin's lymphoma. Economic impact of rituximab (Mabthera) versus conventional chemotherapy]. AB - The monoclonal antibody rituximab, targeted against the CD20 antigen, has shown efficacy in patients with follicular lymphoma who relapse or fail to response to conventional chemotherapy. We evaluated the economic impact of using rituximab for the treatment of non-Hodgkin's lymphoma (NLH) in comparison with conventional chemotherapy protocols (CHOP or CHVP). In this retrospective study conducted between 1998 and 2000, the direct costs of treating inpatients with NHL rituximab (n=20) or CHOP/CHVP (n=17) were compared. Results, including costs of administering chemotherapy and adverse events, showed that the average cost per patient was comparable for the two strategies (9700 euro for rituximab, versus 8487 euro for conventional chemotherapy). In the rituximab group, the cost was mostly due to drug purchases. In the conventional chemotherapy group, outlays were related to drug-induced toxicity and longer hospital stay. Our results were similar to others described in the literature. Prospective studies are nevertheless needed for confirmation. For first-line treatement, the difference in the cost-effectiveness-ratio between rituximab and conventional drugs might be smaller, but sound data are not yet available. PMID- 12910042 TI - [Pathophysiology of Sjogren's syndrome]. AB - Sjogren's syndrome (SS) is an excellent model for understanding the pathophysiology of autoimmune diseases and the relationships between autoimmunity and lymphoma. Recently discovered new elements probably play a role in the pathogenesis of this multifactorial disease: genetic predisposition remains largely unknown, but there isa link between certain HLA molecules and the type of autoantibodies secreted; sometimes called autoimmune epithelitis, SS is associated with abnormal apoptosis activity in epithelial cells leading to an abnormal accumulation of degradation products of the cytoskeleton proteins such as alpha- and beta-fordrine and also to the presentation of numerous antinuclear autoantigens to the immune system; significant polyclonal activation of B lymphocytes is probably mediated, at least in part, by a major increase in molecules of the TNF family (e.g. BlyS or BAFF) which play an important role in the production of autoantibodies; cytokine inhibition of healthy glands or anti muscarin receptor antibodies and abnormal function of certain water pumps such as aquaporine could explain the perturbed function of the remaining healthy glands; permanent stimulation of autoreactive B cells favors oncogenic events and could lead to the development of B lymphoma with autoantibody activity. The links between these different elements are progressively falling into place. A better understanding of the pathophysiology of SS can be expected to lead to the development of much needed new therapeutic tools. PMID- 12910043 TI - [Sarcoidosis of the kidney and skin without lung involvement]. AB - We report the case of a patient with predominantly renal sarcoidosis. Renal failure responded well to systemic corticosteroid therapy. The clinical presentation was particular, notably the absence of lung involvement and the presence of cutaneous lymphedema. PMID- 12910041 TI - [Transposition of an American-designed comprehensive medical student examination within the framework of the forthcoming French nationwide comprehensive examination. A preliminary study]. AB - Medical training is undergoing extensive revision in France. A nationwide comprehensive clinical competency examination will be administered for the first time in 2004, relying exclusively on essay-questions. Unfortunately, these questions have psychometric shortcomings, particularly their typically low reliability. High score reliability is mandatory in a high-stakes context. The National Board of Medical Examiners-designed multiple choice-questions (MCQ) are well adapted to assess clinical competency with a high reliability score. The purpose of this study was to test the hypothesis that French medical students could take an American-designed and French-adapted comprehensive clinical knowledge examination with this MCQ format. Two hundred and eighty five French students, from four Medical Schools across France, took an examination composed of 200 MCQs under standardized conditions. Their scores were compared with those of American students. This examination was found assess French students' clinical knowledge with a high level of reliability. French students' scores were slightly lower than those of American students, mostly due to a lack of familiarity with this particular item format, and a lower motivational level. Another study is being designed, with a larger group, to address some of the shortcomings of the initial study. If these preliminary results are replicated, the MCQ format might be a more defendable and sensible alternative to the proposed essay questions. PMID- 12910044 TI - [Non-metastatic intrahepatic cholestasis associated with bronchial adenocarcinoma]. AB - In 1961, Stauffer first described a syndrome characterized by nonmetastatic intrahepatic cholestasis associated with undifferentiated renal adenocarcinoma. Since that time, this syndrome has been associated with other tumor diseases. We describe here a patient with lung adenocarcinoma which led to paraneoplastic cholestasis. We discuss the diagnosis and review the literature, emphasizing the pathophysiology of Stauffer's syndrome. PMID- 12910045 TI - [Gastrointestinal bleeding revealing polycythemia. A case report]. AB - PURPOSE: Gastrointestinal bleeding, an uncommon complication of polycythemia can be the inaugural sign in exceptional cases. CASE REPORT: A 35-year-old patient was hospitalized for upper gastrointestinal bleeding. Physical examination and laboratory tests led to the diagnosis of polycythemia (Vaquez disease). Gastroscopy showed an hemorrhagic bulber ulcer. Outcome was favorable after chemotherapy and antinuclear treatment. CONCLUSION: Based on this observation and data in the literature we propose that this unusual presentation would suggest a relationship between polycythemia and ulcer disease. PMID- 12910046 TI - Aseptic meningitis associated with intravenous administration of dexchlorpheniramine. AB - Drug-induced aseptic meningitis has been reported mainly with the use of nonsteroidal anti-inflammatory drugs, antibiotics, intravenous immunoglobulins and OKT3 antibodies. We describe today a very unusual reaction on intravenous dexchlorpheniramine with this case of aseptic meningitis. PMID- 12910047 TI - [Hypersensitivity to celecoxib]. AB - Celecoxib is a non-steroidal anti-inflammatory drug (NSAID) which acts via specific inhibition of cyclooxygenase-2 (synthesis of prostaglandins mediating pathological inflammation) but which preserves the homeostatic action of cyclooxygenase-1. The efficacy of celecoxib is similar to that of classical NSAIDs and its tolerance is comparable with that of placebo. We nevertheless observed one patient with hypersensitivity to celecoxib who developed fever, toxic skin reaction, and liver involvement. PMID- 12910048 TI - [Spontaneous hematoma of the iliac psoas muscle in chronic myeloid leukemia. A case report]. AB - Spontaneous hematoma in the ilio-psoas muscle is an uncommon condition, usually observed as a complication of anticoagulation or hemophilia. Clinically, the onset is marked by violent pain in the territory of femoral nerve and/or psoitis. The diagnosis is confirmed by echography or CT-scan. The most serious complications are loss of self-sufficiency and neuro-muscular after-effect. Surgery is recommended in patients with neurological suffering, followed by early physiotherapy. We report the case of a 42-year-old man, with an ilio-psoas muscle hematoma, revealing chronic myeloid leukemia, without any hemostasis disorder. PMID- 12910049 TI - [Peroneal nerve palsy induced by anterior tibialis pyomyositis]. AB - A 16-year-old girl from Guadeloup developed paresis of the flexors of the right foot, associated with edema and acute pain located in the upper anterior tibialis muscle. Electromyography confirmed mononeuritis of the right peroneal nerve, with severe reduction of potential amplitude. Computed tomography of the right leg showed a heterogeneous mass involving the upper segment of the anterior tibialis muscle, close to the location of peroneal nerve. Muscle biopsy confirmed pyomyositis. Muscle culture was negative. Paresis improved soon after antibiotic therapy was started. PMID- 12910050 TI - [Cephalic tetanus and Bell's palsy in an elderly man]. AB - Cephalic tetanus is a rare form of tetanus defined as trismus plus paralysis of one or more cranial nerves. The seventh cranial nerve is most commonly involved. The process can remain localized, or become generalized. We describe the case of an elderly Caucasian man who developed cephalic tetanus after head trauma with a facial wound. Bell's palsy was inaugural. PMID- 12910051 TI - [Can we foresee type 2 diabetes: after the Diabetic Prevention Program]. PMID- 12910052 TI - [The diabetic pregnant woman]. AB - Clinical data demonstrated a close relationship between the conceptional glycated haemoglobin and the occurrence of spontaneous early abortions and fetal malformations. Blood glucose level during pregnancy was correlated with the risk of fetal macrosomia with significant links between birth weight, fetal hyperinsulinemia and mean maternal blood glucose levels. Tight blood glucose control from conception to term was clearly shown to be able to lower the risk of fetal abnormalities and perinatal mortality to that of the offspring of a non diabetic mother. But this imply mainly to plan the pregnancy and to maintain a tight normoglycemia from the conception, and to establish a close partnership between diabetologist and obstetrician. The diagnosis and the management of gestational diabetes mellitus (GDM) are still subject to a large debate, and particularly screening and diagnosis criteria. However, diagnosis and management of GDM are clearly mandatory, particularly to identify the undiagnosed type 2 diabetic women. As well as for a diabetic pregnancy, the therapeutic management of GDM must be instituted early and must be intensive in order to reduce the level of fetal macrosomia to the one of the non-diabetic population. PMID- 12910053 TI - [Maturity-onset diabetes of the young (MODY): the history of its dismemberment]. AB - Type 2 diabetes (T2D) is a complex multifactorial disease. Genetic susceptibility is clearly needed, but in most cases, is not sufficient to induce T2D. Environmental factors are determinant in most patients to develop T2D. Monogenic forms of diabetes such as the maturity-onset diabetes of the young (MODY) represent a model to study the genetic basis of T2D. Studies in MODY individuals and families lead to a genetic definition of MODY subtypes with profound implications for our understanding of gene mutations involved in the development of T2D through our understanding on the major role of these genes and on their transcriptional activity on b-cell development and function; and on the regulation of glucose and insulin metabolism. Our current knowledge on the heterogeneity of MODY diabetes and contribution to our comprehension of the molecular mechanisms involved in T2D are described. PMID- 12910054 TI - [MODY in clinical practice. When and why to consider it?]. AB - It is important to consider in clinical practice the diagnosis of MODY and to characterise the MODY sub-type. Indeed, evolution, clinical consequences and treatment of these forms of diabetes will be different according to the genetic origin of MODY. The main characteristics and consequences in clinical practice of these genetic abnormalities are briefly described, with a particular emphasis on the two most frequent sub-types, MODY-2 and MODY-3, caused by mutations of the enzyme glucokinase and of the transcription factor hepatocyte nuclear factor-1 alpha (HNF-1a) respectively. PMID- 12910056 TI - [To observe, or to observe and to act?]. AB - The UKPDS Study has clearly shown that diagnosis and therapeutic management of type 2 diabetes and its complications occur lately in the course of the disease, and that diabetes control deteriorates and micro- and macrovascular complications risk increase with the duration of the disease. Recent data from the STENO-2 study demonstrate the cardiovascular benefit of an intensified multifactorial management of diabetes and associated cardiovascular risk factors in type 2 diabetic patients at high cardiovascular risk. This paper advocates for an early intensive management of type 2 diabetes from the diagnosis in order to maintain glycaemia and glycosylated haemoglobin at the lowest possible level (even less than the current recommended levels) and for an early aggressive management of all known risk factors (including overweight and obesity, elevated arterial blood pressure, dyslipidaemias, etc.). This implies the early use of multiple therapy at adequate doses and a dramatic change in conventional medical habits, from a late and delayed management of diabetes and its complications to their early prevention using all therapeutic resources. PMID- 12910057 TI - [Metabolic syndrome or insulin resistance syndrome. Recent epidemiological data]. AB - Three definitions are currently proposed to define metabolic syndrome or insulinresistance syndrome (WHO, EGIR, NCEP-ATP III). These definitions are described and recent epidemiological data assessing the frequency of this syndrome in different populations according to these three definitions are presented. Observational studies indicates an increased cardiovascular morbidity and mortality risk in subjects with metabolic/insulin resistance syndrome. These findings imply to identify these at risk subjects and to define the optimal preventive management strategy. PMID- 12910055 TI - [The acceptance of the diabetic disease]. AB - Diabetes is a chronic disease and the diabetic patient will have to live with it. He/She will have to accept this fact and to accept the disease. Several adaptative phases will occur during the course of the disease, linked to the disease evolution. Regular contacts with various members of the healthcare team will have to be positive, both in terms of the patient and healthcare individuals. Adherence and compliance to the treatment and long-term follow-up of the disease will have to be envisaged with a practical positive aspect. Main points and psychosocial aspects are discussed in this paper. PMID- 12910058 TI - [Is it possible to prevent type 2 diabetes?]. AB - Four prospective randomised long-term studies have been recently completed and published (Diabetes Prevention Study, Diabetes Prevention Program, STOP-NIDDM trial, XENDOS Study). Both of them clearly demonstrate the possibility to delay and/or prevent the onset of type 2 diabetes in at high-risk subjects with impaired glucose tolerance, through changes in lifestyle (dietary intervention, weight reduction, increased physical activity) or drug treatment (metformin, acarbose, orlistat). These studies are reviewed and practical implications of their results are discussed. PMID- 12910059 TI - [Screening for diabetic retinopathy: non-mydriatic retinal photography]. AB - The screening of diabetic retinopathy is still largely insufficient and consequently, its management is often delayed. The various recommendations indicate a fundus examination at diabetes diagnosis and yearly thereafter. In France, this is far to be reached, and this situation is at risk of aggravation with the increasing number of diabetic patients and the decreasing number of ophthalmologists. New efficient tools, as the non-mydriatic cameras and digital photography, may allow a new network organisation for screening of diabetic retinopathy, with local screening sites linked to a centralised ophthalmological reading center where digital images could be transmitted. The various aspects of this screening and possibilities to improve the screening and early management of diabetic retinopathy are discussed in this paper. PMID- 12910060 TI - [Pilot study and preliminary validation of the French version of a quality-of life questionnaire specific for subjects with growth hormone deficiency]. AB - This study forms part of a research project seeking to develop a standardized questionnaire by which clinicians can assess the impact of growth hormone (GH) deficiency and its treatment on the "perceived health" or health-related quality of life of adults. The specific aim of this study was to translate and adapt for French patients the AGHDA (Adult Growth Hormone Deficiency Assessment) a standardized health-related quality of life measure for use with GH-deficient adults, initially developed in the United Kingdom, and to collect data which could be used to assess the main psychometric characteristics of its French version the ISPA-HC (Indicateur de Sante Perceptuelle Adulte-Hormone de Croissance). The main properties analyzed are: 1/ The scale's acceptability, as determined by means of face-to-face interviews with a small number of subjects, then by an ad hoc questionnaire administered during a test-retest study; 2/ The scale's reliability, as determined by a test-retest study (with a 15-days interval between tests); 3/ The scale's concurrent validity, as expressed by comparison with scores obtained by means of a generic quality of life scale, the ISPN (the French version of the Nottingham Health Profile). The results of this first trial with the ISPA-HC are conforming to what one can expect from a good instrument. The ISPA-HC has been shown to have very good levels of reliability and internal consistency. Its scores show a close correlation with those of the ISPN (the French version of the Nottingham Health Profile). This instrument can be used to measure variations in the perceived health of subjects with growth hormone deficiency. Its responsiveness to change is to be examined in subsequent studies. PMID- 12910062 TI - Adrenal insufficiency and diabetes mellitus secondary to the use of topical corticosteroids for cosmetic purpose. AB - We report a case of symptomatic topical corticosteroid-induced adrenal insufficiency and diabetes in a 46-yr old HIV 1 positive woman of African descent. Topical Betamethasone dipropionate 0.05%-containing creams were used for the purpose of bleaching over a 2 month period prior to the acute episode. She recovered from her acute onset diabetes with ketosis and adrenal insufficiency a few months after withdrawal of corticosteroids. Despite possible discussion about pathophysiology of diabetes because acute-onset remitting diabetes is not rare in patients of African descent, and diabetes may occur in patients taking anti retroviral treatments, no other cause of a hypothalamo-pituitary-adrenal axis disorder was found. This case suggests that chronic use of high dose topical corticosteroid containing creams should be ruled out in patients presenting with Hypothalamo-Pituitary-Adrenal hypofunction. PMID- 12910061 TI - [Reninoma: a rare but curable cause of high blood pressure, a case report]. AB - We report a case of a renin secreting tumor, which is a very rare cause of secondary high blood pressure. A 22-year-old woman was hospitalised for exploration of high blood pressure (160/110 mmHg) with severe hypokaliemia (2,7 mmol/l) and secondary hyperaldosteronism. Physical examination was normal except the high blood pressure. Bioassays show increased kaliuresis (66 mmol/24h), plasma renin (89 pg/ml in clinostastism--108 pg/ml in orthostatism), pro-renin (1207 pg/ml in clinostastism--1412 pg/ml in orthostatism) and aldosterone (210 pg/ml in clinostastism--566 pg/ml in orthostatism). The rest of the endocrine tests were normal (cortisol and ACTH at 8:00 am, urinary free cortisol, overnight 1 mg dexamethasone suppression test). Doppler ultrasound method, performed by an experienced radiologist, did not show renal artery stenosis. Abdominal computerized tomography showed a nodular formation at the upper pole of the right kidney, isodense to renal medullary. The size tumor was 15 mm. The renal vein sampling shows high values of renin on both sides whereas, for the pro-renin, the values were higher on the tumor side. In spite of treatment with CEI (Converting Enzyme Inhibitors) and calcium antagonists, the blood pressure was not controlled. Hypokaliemia persisted (3 mmol/l) in spite of high daily potassium intake (64 mmol/l of potassium chloride). After tumor resection, reninoma was diagnosed by the pathology examination and blood pressure, plasma rennin, plasma aldosterone level returned to normal. PMID- 12910063 TI - [Werner's syndrome and endocrine disorders]. AB - Werner's syndrome is a rare autosomal recessive disease caused by the mutation of DNA helicase gene (WRN), characterized by the premature onset of multiple age related disorders and skin changes similar to those observed in scleroderma. Some endocrinologic and metabolic disorders have been described in patients with Werner's syndrome. We report one case in a 41-year-old man issuing from consanguineous parents, who presented for exploration of hypoglycemic episodes and sexual impotence. Werner's syndrome was diagnosed on the basis of his characteristic clinical appearance. Metabolic disorders were insulin-requiring diabetes and hypertriglyceridemia. Endocrinologic investigation revealed nodular goiter, sub clinical primary hypothyroidism, hypergonadotrophic hypogonadism,adrenal cortical hypofunction and GH deficiency. Pathology examination of the skin biopsy showed a scleroderma-like aspect. Finally, osteoporosis, atherosclerosis and sub-capsular cataract were associated. Thus, in Werner's syndrome metabolic and endocrinologic investigation is necessary in order to treat these disorders and improve the patient's prognosis and life. PMID- 12910064 TI - Reducing the number of T3 orders in the Paris hospital network: towards better appropriatness of thyroid function test prescription. AB - In vitro thyroid function tests are among the most frequently prescribed laboratory procedures. Serum triiodothyronine (T3) tests are seldom necessary as a first-level measurement. Our objectives were to measure the proportion of T3 measurements relative to all in vitro thyroid function tests in a large hospital network and to investigate the contributions of various interventions to change prescribers'behavior. We performed two cross-sectional surveys in 1995 and 1998 in the 50 Paris University hospitals. Questionnaires were mailed to the heads of the 30 laboratories performing thyroid function tests. One-month orders of free and total thyroxine, free and total T3 and thyrotropin were recorded; changes in T3 measurement orders between the two periods were estimated and association with interventions were expressed as odds ratios and 95% confidence intervals. Twenty five heads of laboratory responded to both surveys. In 1995, T3 measurements constituted 21% of in vitro thyroid function test ordering, which seems to us exceedingly high. The decrease in T3 measurement ordering observed in 1998 (15% of thyroid function test ordering) was independently associated with multiple behavioral changes: educational interventions, structured test form use and year of prescription. PMID- 12910065 TI - [Effects of a 19-norprogesterone derivative, the fourth decade nomegestrol acetate, on lipids]. AB - Dyslipidemia is a cardiovascular risk factor which commonly develops during forty. In Europe, progestins are frequently prescribed for treatment of perimenopausal symptoms in women in this age group, as well as in combination with estrogen replacement therapy in non hysterectomised postmenopausal women. Their complete metabolic tolerance is an important, even if non exclusive, factor to take in consideration for cardiovascular protection. Our aim was to review available data on the effects of a 19-norprogesterone derivative, nomegestrol acetate, on lipid tolerability. In healthy or at risk premenopausal women, clinical studies found no significant changes in lipid parameters (total, HDL and LDL cholesterol, triglycerides, apoprotein B, Lp(a) and LpA-I) with nomegestrol acetate administered in antigonadotropic sequence, alone or combined with estrogen in inverse sequence, during 6 to 9 cycles; there was only a small statistically significant decrease in apoprotein A1, probably due to the induced hypoestrogeny. In clinical studies carried out in postmenopausal women, nomegestrol acetate combined with estrogen replacement therapy in a sequential or continuous combined regimen, did not alter the beneficial estrogen-induced lipid profile: reductions in total and LDL cholesterol, apoprotein B and Lp(a); HDL cholesterol was unchanged and an increase in triglycerides occurred only with oral estrogens. A decrease in apoprotein A1 was found after six months of a cyclic sequential hormone replacement therapy but was associated with a beneficial increase in LpA-I. Nomegestrol acetate has proven its neutral effects on lipid metabolism and does not alter the beneficial estrogen-induced lipid effects. PMID- 12910069 TI - [Vitamin E and neurodegenerative diseases]. PMID- 12910066 TI - [Two cases of non-functional gonadotroph adenoma pituitary apoplexy following GnRH-agonist treatment revealing gonadotroph adenoma and pseudopituitary apoplexy after GnRH administration]. AB - We report here two cases of pituitary apoplexy or pseudoapoplexy revealing a gonadotroph adenoma. A 69-year-old man, who had just started antiandrogen treatment (Gn-RH agonist) for prostatic cancer, was admitted to neurosurgery emergency because of increasing headache and visual impairment. The CT-scan disclosed the presence of a large pituitary mass with lateral invasion of the left cavernous sinus. Hormonel testing showed panhypopituitarism. A few days later, diabetes insipidus appeared. The patient first received corticosteroid therapy and underwent surgical adenomectomy. Immunostaining of the tumor tissue was positive for FSHbeta, confirming the diagnosis of gonadotroph adenoma. Three months after surgery, the endocrine evaluation showed pituitary insufficiency. An 81-year-old man complained of mnemonic disorders. The CT-scan revealed a pituitary mass without extension. The Ophthalmological examination showed left temporal upper quadranopsia. Endocrinological tests with administration of GN-HR triggered headache and vomiting. A second CT-scan was unchanged. Hormone testing revealed increased serum levels of FSH and decreased serum levels of LH. Surgical management of the primary tumor was undertaken due to the visual field alteration. Immunohistochemical studies confirmed the diagnosis of gonadotroph FSHbeta adenoma. PMID- 12910067 TI - [Ciprofibrate-induced acute cholestatic hepatitis]. PMID- 12910070 TI - [Dysembryoplastic neuroepithelial tumors]. AB - Dysembryoplastic neuroepithelial tumors DNTs are highly polymorphic tumors that arise during embryogenesis. They are preferentially, but not exclusively, located in the supratentorial cortex. Histologically they may mimic any categories of low grade or even of high-grade gliomas, but from a carcinological point of view, they behave as stable lesions. Their differential diagnosis from gliomas is obviously important to spare these young patients with a normal life expectancy the long- term deleterious effect of radiation or chemotherapy. The diagnosis of DNT must be considered when all the following criteria are present: partial seizures with or without secondary generalization, no neurological deficit or a stable congenital deficit, cortical topography on MRI, absence of peri-tumoral edema and of mass effect. In other locations, the diagnosis of DNT has to be suspected in case of discordance between the neurological status of the patient and the topography of the tumor or of unusual radiological features such as contrast enhancement but no mass effect and no edema. Supratentorial cortical DNTs tend now to be detected more systematically by imaging soon after first seizures. In most instances, the epilepsy can be cured by gross total surgical removal. Surgery also allows to prevent the risks of intratumoral hematoma or infarct. DNTs should therefore be operated soon after diagnosis. However, excellent results can also be obtained by epilepsy surgery in patients with long term drug resistant partial seizures. PMID- 12910071 TI - [Presenting psychiatric and cognitive disorders in adult neurolipidoses]. AB - Neurolipidoses may present as psychiatric illness or dementia which may be isolated for a long time without neurological manifestations. Thus the relation with a metabolic disease may be difficult to establish. In this survey, we wish to present our clinical and biological experience in relation with lysosomal or peroxisomal disorders giving rise to neurolipidoses, a review of the literature, as well as the elements which allow to present a diagnostic strategy. We report mainly on metachromatic leukodystrophy, GM2 gangliosidosis, Fabry's disease, Niemann-Pick type C, Kufs disease, adrenoleukodystrophy, cerebro-tendinous xanthomatosis. Psychiatric symptoms may overshadow subtle signs of cognitive and motor dysfunction. Careful and persistent neurodiagnostic evaluation must be performed even in cases when CT and MRI scans are considered normal. Resistance to psychotropic drugs may be an element of orientation. The biological diagnosis is mainly biochemical. Although most of the genes involved have been cloned, many of the mutations are private, except for metachromatic leukodystrophy for which specific mutations may be related to adult cases and either with predominantly motor or predominantly cognitive and psychiatric manifestations. This review discusses also other metabolic diseases which may present as isolated or predominant cognitive and psychiatric manifestations. PMID- 12910072 TI - [Intravenous immunoglobulins for relapsing-remitting multiple sclerosis after failure of treatment with other immuno-modulators]. AB - Intravenous immunoglobulins are used in the treatment of different autoimmune diseases. Recent trials suggest their efficacy in relapsing remitting multiple sclerosis. We report the results of an efficacy and safety trial using monthly intravenous injections of immunoglobulins for patients with secondary progressive multiple sclerosis. Eighteen patients in clinical progression, who have been previously treated with immunomodulatory or immunosuppressive drugs, were given monthly intravenous immunoglobulin infusions (0.4 g/kg/d for 5 days). At the beginning, the mean EDSS score was 6.77. At the end of the study, an improvement of EDSS was noted in 61.1 p. cent of patients, with less than 1 and 0.75 for secondary and primary progressive diseases respectively. No worsening was reported. Surprisingly, some patients had partial improvement of neurological functions which were considered as sequelae. Indications for intravenous immunoglobulins in the treatment of the multiple sclerosis need to be evaluated. PMID- 12910073 TI - [Primary dural lymphoma: a report of two cases with review of the literature]. AB - Primary lymphoma arising in dura is exceedingly rare. We report the clinicopathologic findings of two patients with primary B-cell lymphoma of dura. Both were female, 38 and 45 years old. Prior to biopsy they were felt to have meningioma on preoperative magnetic resonance imagery. Histologically, tumors were classified as MALT-type lymphoma. Literature describe only 14 reports of similar entity. Primary lymphomas arising in dura appear to have a more favourable clinical course compared to PCNSL and may require a less aggressive treatment. PMID- 12910075 TI - [Disorders of the processing of spatial information in patients with right cerebral lesions and left hemi-neglect]. AB - The aim of this research was to identify, analyze and classify disorders in behavior which occur in the use of mirrors in patients with right cerebral damage presenting left visual spatial hemiplegia. This work was based on models of visual information processing. Seven controls and eleven patients with right cerebral damage performed a test involving grasping of an object using only specular information from a conventional mirror and then from an inverted mirror. The controls grasped up all the cubes straight away. They only experienced minor difficulty with the inverted mirror, mainly in relation to lateral displacement. The patients revealed a variety of behaviors: 1) searching for and trying to seize the object in the conventional and/or inverted mirror, 2) inversion of the paralysed side (left versus right) in the inverted mirror or the appearance of a visual spatial hemiplegia, 3) modifications in the order of grasped (from right to left, from left to right, or at random), 4) directional anomalies in the horizontal plane linked (or not) with disorders in the use of the anteroposterior space. The results of this study confirm that the patients have abnormal behavior in mirror spaces. While the characteristics of this behavior shows analogies with those described in the non-recognition of objects and/or defects in the processing of visual information for localising objects in space, they can be disassociated from them, and constitute separate syndromes. Specific terminology and taxonomy for the clinical forms of mirror agnosia and specular agnosia, of mirror paralysis and specular paralysis, and of specular ataxia are proposed. PMID- 12910074 TI - [Peripheral haemosiderosis of the central nervous system]. AB - We report 2 cases of patients with hemosiderosis of the central nervous system. The diagnosis was made in 1999 in a stroke unit. The patients had both deafness, but their clinical presentation was different, due to heterogeneous features of this pathology. Hemosiderosis of the central nervous system is due to chronic subarachnoidal hemorrhage. Magnetic resonance imaging is extremely sensitive to the presence of hemosiderin and is the investigation of choice enabling diagnosis and sometimes bleeding source (50 p. cent). The interest of our observations concerned the source of bleeding. In the first patient, diagnostic work-up was negative. For the second one, the deposition of hemosiderin was explained by 3 possible sources. If a bleeding source can be identified, surgery is the optimal treatment. Copper chelation, like Trientine can be used, but there is no evidence based for using this treatment. PMID- 12910076 TI - [Diffusion tensor MRI of Wallerian degeneration: a case report]. AB - Diffusion tensor imaging allows a 3D analysis of water molecular motion and an exploration of white matter tracts integrity and orientation. We report a diffusion tensor study of wallerian degeneration of the corticospinal tract following a capsulo-lenticular hemorrhage in a 49-year old man. Reduced anisotropy (loss of fiber coherence) with preserved diffusivity of the damaged corticospinal tract was observed. This highlights the feasibility of diffusion tensor imaging on clinical MR units and its ability to quantify the degree of white matter tract destruction. PMID- 12910077 TI - [Hashimoto's thyroiditis associated with recurrent transverse myelopathy]. AB - A 55-year-old woman presented first episode of paraplegia at D6 level with a marked improvement of neurological signs, but relapse occurred five months later. Hashimoto's thyroiditis with euthyroidism was found. Long-term 9 months treatment with oral corticosteroids was applied. Also she was able to walk with help. Two years after the second palsy she developed a new paraplegia at D6 level with a severe residual deficit five months later in spite of oral corticosteroids. Thyroid antibodies remained positive. Devic's disease without optica neuritis associated with thyroiditis or spinal cord localisation of Hashimoto's encephalopathy was discussed. PMID- 12910078 TI - [Camptocornia presenting with a proximal myotonic myopathy]. AB - We report the case of a 74-year-old patient who presented with an anterior inflexion of the trunk which increased during the day. His past medical history included treatment for hypothyroidism, a cure of cataracts and an increase of gammaGT. This camptocormic attitude revealed a proximal myotonic myopathy (PROMM). Clinical and paraclinical arguments (hypothyroidism, cataracts, weakness, EMG, muscle biopsy, biology) led to diagnosis. PMID- 12910079 TI - [Glycogenesis due to adult phosphorylase kinase deficiency]. AB - A 42-year-old man presented exercise-induced muscle pain without myogloburia since the age of 12 years. Histochemistry and electronmicroscopy of a muscle biopsy revealed subsarcolemmal and inter-myofibrillar accumulation of glycogen. Exercise on a bicycle ergometer produced a normal raise of lactate. Biochemical study showed a partial defect in phosphorylase activity. PMID- 12910080 TI - [Bilateral amyotrophic neuralgia (Parsonage Turner syndrome) with HIV seroconversion]. AB - We report a unusual case of neuralgic amyotrophy (NA) occurring during the seroconversion stage of an HIV infection. Combined with previously published cases, our observation suggests that NA associated with HIV could belong to the group of early multiplex mononeuritis. Neurologists should be aware of HIV infection when managing a patient with NA. PMID- 12910081 TI - [Two unusual complications of the Klippel-Trenaunay syndrome: carpal tunnel syndrome and cerebral venous thrombosis]. AB - Most neurological complications of the Klippel-Trenaunay syndrome are due to the presence of cerebral or spinal arteriovenous malformations and their consequences. We report 2 cases of unusual complications of this syndrome. In the first case, where the anomalies were restricted to the left upper extremity, the occurrence of hand numbness led to the discovery of a carpal tunnel syndrome, probably due to the lymphatic abnormal proliferation within the carpal tunnel. In the second case the disease was much more widespread and affecting, among other localizations, the cephalic extremity. The patient presented with intractable left motor seizures that appeared to be due to an occlusion of the right transverse and sigmoid sinuses. Treatment with heparin and anticonvulsants led to rapid recovery. PMID- 12910082 TI - [Gait disorders with insidious progression in a 60-year old woman]. PMID- 12910083 TI - [A leucoencephalopathic presentation of cerebral lymphoma]. PMID- 12910084 TI - [Methodological issues relevant to clinical research in rare diseases]. PMID- 12910085 TI - [Pneumology Society of the French Language. Guidelines for the clinical management of COPD]. PMID- 12910086 TI - [Guidelines for the clinical management of COPD, 2003 update: organisation and argumentation]. PMID- 12910087 TI - [Guidelines for the clinical management of COPD. Definition, nosology and natural history]. PMID- 12910088 TI - [Guidelines for the clinical management of COPD. Epidemiology]. PMID- 12910089 TI - [Guidelines for the clinical management of COPD. Risk factors]. PMID- 12910090 TI - [Guidelines for the clinical management of COPD. Diagnosis, severity, organization of follow-up]. PMID- 12910091 TI - [Guidelines for the clinical management of COPD. Prevention and reduction of risk factors]. PMID- 12910092 TI - [Guidelines for the clinical management of COPD. Bronchodilatators]. PMID- 12910093 TI - [Guidelines for the clinical management of COPD. Corticosteroids]. PMID- 12910094 TI - [Guidelines for the clinical management of COPD. Other drugs]. PMID- 12910095 TI - [Guidelines for the clinical management of COPD. Rehabilitation]. PMID- 12910096 TI - [Guidelines for the clinical management of COPD. Oxygen therapy and long-term ventilatory assistance]. PMID- 12910097 TI - [Guidelines for the clinical management of COPD. Surgical treatment]. PMID- 12910098 TI - [Guidelines for the clinical management of COPD. Exacerbations/acute respiratory failure: diagnosis, severity and management]. PMID- 12910099 TI - [Guidelines for the clinical management of COPD. Exacerbations/acute respiratory failure: antibiotherapy]. PMID- 12910101 TI - [Bronchial obstruction and dyspnea: detection, perception, sensation]. PMID- 12910102 TI - [How to target biotherapies of lung cancer? Bronchoscopic biopsy and the immunohistochemical detection of the EGF-receptor]. PMID- 12910103 TI - [Reducing the risks associated with bronchoscopy: a call for a standardisation of practice]. PMID- 12910104 TI - [Sudden infant death syndrome: "sleep tight on your back..."...and what else?]. PMID- 12910105 TI - [Health effects of air pollution episodes]. PMID- 12910106 TI - [Asthma and sports: danger!]. PMID- 12910107 TI - [Tolerance of fiberoptic bronchoscopy by self-administered questionnaire: in the words of the patients]. AB - INTRODUCTION: Fibreoptic bronchoscopy is currently undertaken by the majority of respiratory physicians, but under varying conditions. Though complications are rare the tolerance of this examination is sometimes poor, particularly when it is performed under local anaesthesia. The undesirable effects may reduce the value of the examination as well as causing discomfort for the patient. METHODS: A prospective study of the tolerance of the endoscopic examination was made on 100 consecutive patients by self-administered questionnaire. RESULTS: There were no major and 7 minor complications (7%). 45% of the patients were anxious but the experience of the operator tended to reassure them (p=0.07). 30% of the patients reported some pain, which tended to be exacerbated by anxiety (44% vs 18%, p=0.008) and the supine position (57% vs 43%, p=0.047). 37% of patients reported nausea, and 50% dyspnoea, without any significant predictive factor. 79% would agree to a repeat examination under the same conditions and 92% said that they had received information appropriate to the examination undergone. CONCLUSION: The tolerance of fibreoptic bronchoscopy under local anaesthesia is poor and perhaps overestimated by respiratory physicians. Patient information is essential. A national enquiry could lead to the standardisation of techniques. PMID- 12910108 TI - [Impact of prior biological assessment of coagulation on the hemorrhagic risk of fiberoptic bronchoscopy]. AB - INTRODUCTION: The goals of the study were to test the value of coagulation tests to predict bleeding events during fiberoptic bronchoscopy (FOB) and to identify risk factors. METHODS: A monocentric prospective study of 426 procedures performed by one physician was realized. A standardized questionnaire and coagulation tests including prothrombine time, activated cephaline time and platelets counts were performed before the procedure. Bleeding events, defined by a loss of more than 50 ml of blood, were recorded for all patients. RESULTS: 44 patients (10.3%) had bleeding events, modifying the procedure in 19 cases. No death occurred following FOB during the study. Among the 17 patients with abnormalities on coagulation test before FOB, only one had significant bleeding. Two risk factors were found as predictors of bleeding events: nose or gum bleeding (OR=4.99, CI (95%) [2.6-9.5]; p<0.001) and left cardiac failure (OR=4.53, CI (95%) [1.7-12.1]; p<0.01). CONCLUSION: This study shows that abnormalities on coagulation tests are not predictive for bleeding events. Nose or gum bleeding and left cardiac failure may be risk factors for bleeding events during FOB. PMID- 12910109 TI - [EGF-R expression correlation between biopsy and surgical specimens of lung carcinoma]. AB - INTRODUCTION: Most of the time, biological parameters are evaluated on tissues obtained on surgical samples of the primary tumour. New approaches in non small cell lung cancer (NSCLC) treatment consist to administer neoadjuvant chemotherapy or anti-EGF-R (epidermal growth factor receptor) drug. METHODS: We assessed the expression of EGF-R by immunohistochemistry on biopsy samples and on the paired resected tumours in 27 patients. RESULTS: The mean percentage of EGF-R positive neoplastic cells was 11% in surgical specimens compared to 28% in biopsy specimens (p=0.02) although a good correlation (R=0.67; p=0.0001) between biopsies and surgical specimens was observed. Furthermore, the positivity (cut off > 1% cells) rate was not statistically different between biopsies (55%) and tumours (48%) (p=0.63). In term of positivity rate, we found 85% concordant results between biopsies and resected tumours, 4% false negative and 11% false positive on biopsies in comparison with the resected tumours. The positive and negative predictive value of the biopsies were respectively 80% and 92%. CONCLUSIONS: The evaluation of EGF-R by immunohistochemistry on biopsies may provide reliable information about non small cell lung cancer EGF-R status. PMID- 12910110 TI - [Impact of medical prescription computerisation on the incidence of adverse drug effects]. AB - INTRODUCTION: Adverse drug effects are a significant public health problem. Prescription errors are responsible for a significant proportion of these adverse effects. METHODS: We have aimed to improve the link between generation of and interpretation of a prescription through computerisation. The prescription sheet, which is generated daily, was organised to allow care planning without the need to re-copy out treatments on the sheet. A prescription aid was available which was based on a core group of drugs commonly used in our respiratory service. The aim of the study was to compare the various types of errors observed during 6 weeks of computerized prescriptions (229 files) to a retrospective series of handwritten prescriptions of the service at an identical time (184 files) the previous year. The case-mix was identical for both analysed periods. RESULTS: The total number of technical prescribing errors in the 1,599 handwritten lines (49.27% error) was significantly higher (p<0.001) than the 1,805 computerized prescriptions lines (42.88% error). The errors of copying (p<0.001), eligibility (p<0.001) and incorrect spelling (p<0.05) were the main sources of error which were significantly reduced by computerisation. CONCLUSION: Computerised prescription is likely to reduce the incidence of prescribing errors and adverse drug effects. PMID- 12910111 TI - [Dyspnoea and perception of airway obstruction]. AB - INTRODUCTION: Dyspnoea, defined as the perception of difficult or laboured breathing, is ideally quantified, using a visual analogue scale or a Borg scale. STATE OF THE ART: In recent years several studies have explored the correlation between dyspnoea and the degree of airway obstruction, in both healthy subjects and patients with asthma or COPD. In these studies dyspnoea was measured in baseline state, following induced broncho-constriction or during application of an external resistive load. Dyspnoea was measured using a previously validated scale or, in some studies, a simple numerical score. The lung function variables most frequently used to evaluate the degree of correlation with dyspnoea were FEV1, and PEF, although other variables were used in some studies. Despite this marked heterogeneity in the methods used, several studies strongly suggested that, in subjects with COPD, perceived subjective improvement after an inhaled bronchodilator agent correlated with improvement of inspiratory variables. In asthmatics, the major finding was that some subjects had a very poor perception of their level of airway obstruction, a potential risk factor for severe exacerbations. PERSPECTIVES: Further normative studies are needed to define how healthy subjects perceive airway obstruction. CONCLUSIONS: The relationship between dyspnoea and lung function in obstructive disorders has to be evaluated separately in asthmatics and in COPD patients. PMID- 12910112 TI - [Normal and abnormal alveolar development]. AB - INTRODUCTION: Alveolar growth predominantly occurs post-natally and is characterised by the multiplication of alveoli, the thinning of inter-alveolar walls, and the maturation of capillary vessels. STATE OF THE ART: Alveolar growth is controlled by numerous factors whose interactions remain poorly understood. Many phenomena can interfere with normal alveolar growth resulting in abnormal development and a reduction in alveolar surface. This is especially the case in premature human neonates, whose lungs are structurally and functionally immature. Oxygen therapy, mechanical ventilation, and airway inflammation may induce alveolar growth disorders in these children, resulting in the development of bronchopulmonary dysplasia. PERSPECTIVES AND CONCLUSIONS: Further studies are needed to improve our knowledge about alveolar growth regulatory mechanisms so that better strategies can be developed to prevent these respiratory complications in premature neonates. PMID- 12910113 TI - [Asthma in athletes]. AB - INTRODUCTION: Numerous recent studies have shown that the risk of developing asthma or exercise-induced asthma is increased in the athletic population, particularly in endurance-trained athletes at national and international level. STATE OF ART: According to the literature, this could be explained by both hyperventilation during exercise and increased airway exposure to inhaled allergens, pollutants and/or cold dry air. However this form of asthma seems to differ from classical asthma. PERSPECTIVES: In the future, the establishment of rigorous controls - via a detailed description of symptoms and documentation of objective measurements such as resting spirometry, bronchial hyperreactivity and reversibility - should allow early detection of respiratory problems in athletes and enable to provide an adequate treatment. CONCLUSIONS: Although asthma and exercise-induced asthma are particularly common among athletes, if appropriately detected and treated, these disorders should not constitute a limiting factor in exercise performance. PMID- 12910114 TI - [Non-invasive ventilation of elderly patients in the intensive care unit]. AB - INTRODUCTION: An increasing number of elderly patients, with acute respiratory failure (ARF) due to e.g. COPD exacerbation or cardiac failure, are being referred to intensive care units for mechanical ventilation. STATE OF ART: NIV can be an effective ventilatory technique in ARF due to a variety of aetiologies. NIV has been shown to decrease endotracheal intubation rates, complication rates and median hospital length of stay. When managing acute-on-chronic respiratory failure, NIV is a first-line ventilatory technique, in the absence of criteria for immediate endotracheal intubation. The efficacy of NIV depends on the expertise of the medical and nursing staff (including physiotherapists) and should always be performed in an appropriate setting. There are very few data about NIV in the elderly but studies, which included patients over 75 years, did not identify different outcomes for this age group. PERSPECTIVES: Future clinical studies on NIV should allow us to better understand which patients will benefit the most from the technique. Certain specific settings in elderly populations, such as in palliative care or when an "non intubation order" has been given by the patient, his family or the medical/nursing staff, are under evaluation. CONCLUSIONS: NIV is effective and well tolerated and it has become a key ventilatory technique in the management of ARF, particularly for elderly patients. PMID- 12910116 TI - [The mean, the median, and their dispersion indices: when and how to use them in a research article?]. PMID- 12910115 TI - [A study of early tracheostomy in patients undergoing prolonged mechanical ventilation]. AB - BACKGROUND: Several complications are associated to mechanical ventilation (MV), especially if the duration of MV is prolonged: nosocomial pneumonias and sinusitis, laryngeal and tracheal complications (such as stenosis or granuloma). Pneumonias, en particular, seem to be associated to an increased mortality risk. Overall, the hospital mortality rate of patients undergoing MV for more than 7 days is 45%. Early tracheostomy could allow to reduce some of these complications; however, the actual value of this procedure has never been proven. AIM OF THE STUDY: The aim of this multicenter randomized trial is to assess the potential interest of early tracheostomy (before the 4th day of MV) compared to translaryngeal intubation, in critically ill patients undergoing MV for more than 7 days, in reducing mortality rate, incidence of pneumonias, and duration of MV. METHODS: Inclusion criteria are: MV since<4 days, forseable duration of MV>7 days, age>18-yr, and informed consent obtained. Exclusion criteria are: MV since > 4 days, presence of tracheostomy, major risk of bleeding, cervical infectious disease, "moribund" state (according to SAPS II and OSF scores), MV or CPAP at home, non reversible neurologic disease, intracranial hypertension. The randomization is performed before the end of the 4th day of MV. EXPECTED RESULTS: A decrease of mortality rate on d.28 from 45% to 32% is expected (two-sided test, alpha=0.05, B=0.80); overall, 468 patients should be included over a 3-yr period in at least 50 centers. In addition, a decrease of the incidence of pneumonias and of duration of MV during the first 28 days is expected. Secondary endpoints are mortality rate on d.60, hospital mortality, total duration of MV, infectious complications (other than pneumonias), laryngeal and tracheal complications, duration of sedation, duration of stay in intensive care unit, hospital costs, and comfort of patients. PMID- 12910117 TI - [What does "statistically significant" mean?]. PMID- 12910118 TI - [Well differentiated fetal-type pulmonary adenocarcinoma: a tumour with a good prognosis]. AB - INTRODUCTION: Pulmonary tumours resembling the foetal lung are primitive tumours and are rare. They include pneumoblastomas and foetal-type pulmonary adenocarcinoma. Foetal-type adenocarcinomas (FTA) fall into two categories: well differentiated and poorly differentiated. Poorly-differentiated FTA, which tends to occur in older subjects who smoke and who are male, has a guarded prognosis and is classified as a variety of pulmonary adenocarcinoma. CASE REPORTS: Well differentiated FTAs are also called endodermic tumours resembling the foetal lung. These are large tumours, peripherally sited but with endobronchial extension that occur primarily in younger patients who are often non-smoking females. They comprise of tubular glandular structures and morular metaplasia, compact and rounded cellular proliferation's which contain endocrine cells. We have described three cases. CONCLUSIONS: These tumours are important to classify as their prognosis is good if surgical excision is complete. PMID- 12910119 TI - [Isolated pleural metastases from an atypical meningioma]. AB - INTRODUCTION: Less than 2 per 1,000 meningiomas are complicated by extra-cranial metastases. These are most often found in the lung parenchyma, liver or lymph nodes. They almost always occur in anaplastic meningiomas (grade III according to OMS) and much more rarely in atypical meningiomas (grade II). CASE REPORT: We report a case of pleural metastases from a primary frontal atypical meningioma with no other extra-cranial spread. CONCLUSION: Poorly differentiated meningioma presents many morphological and immuno-histochemical similarities to malignant mesothelioma. For this reason the diagnosis of pleural metastase from a meningioma cannot be made without knowledge of the primary meningeal tumour and its histological type. PMID- 12910120 TI - [Low grade pulmonary sarcoma preceding the discovery of a uterine primary]. AB - INTRODUCTION: Low grade pulmonary sarcomas are very rare tumours. We report the case of a low grade sarcoma of the lung occurring two years prior to the finding of a uterine primary. CASE REPORT: Complete surgical excision of a low grade pulmonary sarcoma was performed. The initial search for dissemination was negative. Two years later a follow-up scan discovered a uterine mass as well as a para-aortic shadow that proved to be the primary tumour (low grade uterine sarcoma) and a metastasis. CONCLUSION: This is the second case of a pulmonary metastasis discovered before a primary low grade uterine sarcoma. The first was found during the investigation of a pulmonary sarcoma. The main differential diagnosis to consider is metastatic leiomyosarcoma. In both cases their finding justifies the search for a uterine primary by immunohistochemical examination for oestrogen and progesterone receptors as well as pelvic ultrasound or even magnetic resonance imaging. PMID- 12910121 TI - [Pneumopathy due to isocyanate hypersensitivity: recognition as an occupational disease]. AB - INTRODUCTION: Hypersensitivity pneumopathies (HSP) are listed as number 66 in the list of occupational diseases but this list includes regulatory requirements that are not applicable to isocyanates. The authors report the case of a worker exposed to isophane di-isocyanate (IPDI) during the manufacture of polyurethane foam. CASE REPORT: The diagnosis of HSP was based on the clinical presentation (dyspnoea, fever, crepitant rales) and the complimentary investigations (ground glass appearance on the thoracic CT scan, lymphocytosis in the broncho-alveolar lavage). It was confirmed by trans-bronchial biopsy. Only the functional assessment (airflow obstruction and absence of marked reduction in CO transfer) was atypical. With regard to the role of isocyanates the number of published cases (49 since the first observation in 1976) shows that this aetiology is rare. In the case presented the chronology of symptoms (regression following exclusion and immediate relapse following re-exposure) and the absence of exposure to other agents known to cause HSP confirm the aetiology with near certainty. This case was recognised within the framework of the complimentary procedure that allows for unlisted diseases. CONCLUSION: In order to assist the management and recognition of HSP due to isocyanates there is good reason to raise the awareness of the medical profession and to extend the list of occupational diseases. PMID- 12910122 TI - [Chronic cough after laparoscopic adjustable gastric banding]. AB - INTRODUCTION: Patients are frequently referred for chronic cough. The causes are various. CASE REPORT: We report two cases of chronic cough that occurred after laparoscopic adjustable gastric banding for treatment of morbid obesity. In both cases, the computed tomography scan showed an important oesophageal dilatation. The cough disappeared after the band deflation. CONCLUSION: Oesophageal dilatation after laparoscopic adjustable gastric banding is a new cause to be included in the aetiology of chronic cough. PMID- 12910123 TI - [Bilateral diffuse pneumopathy in a cabinetmaker]. PMID- 12910124 TI - [Multiple pulmonary nodules with halo sign]. PMID- 12910125 TI - [AFSSAPS guidelines for the antibiotic treatment of lower respiratory track infections]. PMID- 12910126 TI - [Chronic obstructive pulmonary disease: an overview]. PMID- 12910131 TI - [Text of recommendations (long version) of theme I: "Pneumology and study of infectious disease"]. PMID- 12910132 TI - [Cystic fibrosis and survival: what interpretation?]. PMID- 12910133 TI - [Respiratory infection in cystic fibrosis: burden of illness in the progression of cystic fibrosis, previous deductions for the microbiological study of bronchial secretions and clinical criteria of infection]. PMID- 12910134 TI - [What is the burden of respiratory infection in the progression of cystic fibrosis]. PMID- 12910135 TI - [Clinical criteria of infection with Staphylococcus aureus and Pseudomonas aeruginosa]. PMID- 12910136 TI - [What previous deductions can be used for optimizing analysis of bronchial secretions in cystic fibrosis?]. PMID- 12910137 TI - [What microbiological criteria define colonization or infection with Staphylococcus aureus and Pseudomonas aeruginosa]. PMID- 12910138 TI - [What microbiological criteria define a colonization or infection with Staphylococcus aureus?]. PMID- 12910139 TI - [What microbiological criteria define a colonization or an infection with Pseudomonas aeruginosa?]. PMID- 12910140 TI - [Treatment of staphylococcal infection in cystic fibrosis]. PMID- 12910141 TI - [Staphylococcus aureus infection: when and how to treat?]. PMID- 12910142 TI - [Strategy of antibiotic therapy by various routes of administration in the initial colonization by Pseudomonas aeruginosa]. PMID- 12910143 TI - [How should initial colonization by Pseudomonas aeruginosa be treated in patients with cystic fibrosis?]. PMID- 12910144 TI - [Strategy of antibiotic therapy in the course of chronic Pseudomonas aeruginosa infection]. PMID- 12910145 TI - [How should chronic Pseudomonas aeruginosa bronchial infection be treated in the course of cystic fibrosis?]. PMID- 12910146 TI - [Management of patients with cystic fibrosis: role of corticosteroid therapy]. PMID- 12910147 TI - [Role of inhaled bronchodilators]. PMID- 12910148 TI - [Role of rhDNase]. PMID- 12910149 TI - [Role of inhaled corticoids]. PMID- 12910150 TI - [Cystic fibrosis and inhaled bronchodilators]. PMID- 12910151 TI - [Role of rhDNase in cystic fibrosis]. PMID- 12910152 TI - [Role of physical therapy in the infant. Role of assistive equipment in the physical therapy of patients with cystic fibrosis]. PMID- 12910153 TI - [Role of respiratory physical therapy in the infant]. PMID- 12910154 TI - [Role of assistive equipment in the treatment of cystic fibrosis]. PMID- 12910158 TI - [The qualities of a good medical article]. PMID- 12910159 TI - [Management of cancer patients by general practitioners. Results of a survey among 422 physicians in Aquitaine]. AB - OBJECTIVE: To have a better knowledge on the role of the general practitioner in the management of cancer patients, their opinion on communications with the specialists and their training in oncology. METHOD: In March 2000, a questionnaire was sent to a random sample of 730 general practitioners in the Aquitaine area. RESULTS: We obtained 440 answers (60%). The general practitioners declared taking care of a mean of 16 cancer patients among their clientele and seeing 5 new cases every year. Two thirds of them decided on the orientation of their patients, case by case. Nearly 70% of the practitioners reported difficulties, notably psychological, in the case of relapses or during the terminal phase. The majority (94%) considered the quality of the information provided by the specialists as correct or good. Globally, the practitioners would prefer to be more involved in the treatment of their cancer patients and 82% would accept to ensure the follow-up of the patients in remission. CONCLUSION: The expectations of the general practitioners defined in this study merit reflection for the development of treatment networks in order to improve their organisation and the collaboration between general practitioners and specialists. PMID- 12910160 TI - [ERAP, a pharmaco-epidemiologic survey on perennial allergic rhinitis in every day medical practice in France]. AB - OBJECTIVE: In order to gain knowledge on the management of perennial allergic rhinitis by the practitioners in France, a survey (Enquete epidemiologique sur la rhinite allergique per-annuelle en pratique quotidienne--ERAP) was developed. METHODS: In the year 2000, 1 346 doctors (including 689 general practitioners) enrolled 3 507 patients consulting for a fall/winter exacerbation of allergic rhinitis. Both doctors and patients filled out a specific questionnaire on the care networks, clinical characteristics, diagnostic modalities and treatment of allergic rhinitis, together with the patients'personal feelings. RESULTS: The patients were bothered on average 6 days a week (5.8 +/- 1.8 days) and 3 months a year (9.2 +/- 7.7 consecutive weeks) by symptoms largely exceeding the ENT area, with frequent ocular symptoms (36.7%), cough (31.9%), sleep disturbances (35.2%) or dyspnea (9.4%). In 70.3% of the patients rhinitis affected their professional life and 90.9% their daily life. The nose was examined in 63.3% of patients and a search for asthma was conducted in 39.3%. In 45.3% subsequent skin prick tests were scheduled and in 53.5% specific IgE measurements and in 33.5% subsequent specialized consultation. Prescribed drugs were mainly oral antihistamines (75.3%) and local glucocorticosteroids (48.5%). Prescriptions were written for a mean duration of 8 weeks. 78.6% considered that the information that they had received was adequate and easy to understand, but 61.1% of patients would have liked to have received more advice. Furthermore, only 48.7% followed their doctor's prescription scrupulously, and 34.3% adapted the prescription. CONCLUSION: Perennial allergic rhinitis indeed impairs quality of life and its management in France is now better understood. PMID- 12910161 TI - [Thrombosis of the superficial dorsal vein of the penis (penile Mondor's phlebitis). The interest of Doppler examination]. AB - INTRODUCTION: Thrombosis of the superficial dorsal vein of the penis (TSDVP) is a frequent and benign pathology that is often underestimated. OBSERVATION: A patient consulted for acute erectile dysfunction and pain in the base of the penis due to a TSDVP, which was treated with non-steroidal anti-inflammatory agents and anticoagulants and was followed-up by Doppler ultrasound examination of the penis. CONCLUSION: Essentially clinical, the diagnosis of TSDVP can benefit from Doppler ultrasound of the penis in difficult cases, because it is a useful tool for correct diagnosis and therapeutic management of the pathology. PMID- 12910162 TI - [Absence of cutaneous complications following extravasation of pegylated liposomal doxorubicin]. PMID- 12910163 TI - [Streptococcus suis meningitis following wild boar hunting]. PMID- 12910164 TI - [Vancomycin-induced linear IgA bullous dermatosis]. PMID- 12910165 TI - [Urinary disorders in ageing men. A turning point of life]. PMID- 12910166 TI - [Anatomo-physiological aspects of the lower urinary tract in men and modifications related to ageing]. AB - MICTURITION AND INCONTINENCE: The lower urinary tract, composed of several organs necessary for micturition, is controlled by the encephalic and peripheral centres, which have a synergic effect and coordinate the lower urinary tract. Continence is ensured by the effect of the adrenergic sympathetic nerve that increases urethral sphincter tonus and maintains low pressure in the bladder during filling. Micturition appeals to the cholinergic system mediated by the muscarinic receptors situated in the bladder. AGEING OF THE PROSTATE: Under the influence of dihydrotestosterone and growth factors, ageing of the prostate is characterized by histological modifications leading to hypertrophy, obstructing the flow of urine. AGEING OF THE BLADDER: Electronic microscopy studies have shown the presence of degenerative lesions and, on urodynamic level, are characterised by over or under-activity. Ageing of the peripheral and central nervous centres is morphological and metabolic with the production of vascular and free radicals, involving the systems that control micturition and continence. The ageing kidney has a tendency to produce more urine at night because of the functional changes that intervene in the salt and water balance regulation systems. PMID- 12910167 TI - [Clinical manifestations of urinary disorders and their treatment in ageing men]. AB - PROBLEMS OF THE PROSTATE: Benign hypertrophy of the prostate (BHP), when it occurs, is manifested by an obstruction or irritation related to overactivity of the bladder. The obstructive syndrome is defined by urodynamic tests. Urge incontinence and study of the pressure-flow ratio are the tests of choice. The functional handicap and impact on quality of life are assessed using the International Prostatism Symptoms Score (I-PSS). Efficient and fairly well tolerated medical treatment has reduced the indications for surgery. It relies on alpha-blockers, 5a-reductase inhibitors and phytotherapy. When indicated, the surgical treatment of choice is endoscopic resection of the prostate. Among the non-prostatic micturition disorders, urge micturition with, in extreme cases, incontinence are due to detrusor instability. This is of multifactor origin; enhanced by the local irritation or environmental factors, it usually occurs within a context of acute or chronic pathologies. Treatment is recommended with anticholinergic agents. New molecules have recently been launched, better tolerated than oxybutinine. Electrostimulation can be a good alternative in mentally normal patients. Micturition due to excess urine may be due to overactivity of the bladder, the major risk of which is acute urine retention. It can also be observed during neurological affections such as Parkinson's disease or during administration of certain drugs. Nocturnal polyuria is a frequent problem. However, simple hygiene and dietary measures and the control of certain concomitant diseases can usually relieve the symptoms. Medical treatment relies on desmopressine. PMID- 12910168 TI - [The management of waste, a simple question of acceptability?]. PMID- 12910169 TI - [MR-arthrography: a reality in France]. PMID- 12910170 TI - [MR-arthrography: general principles and applications]. AB - The recent approval by the French Ministry of Health of the use of intra articular Gadolinium could promote the increasing use of MR-arthrography in France. Although useful in specific pathologies, it should not be overly prescribed and should be considered only if it provides a more accurate diagnosis than other less invasive techniques. The technical aspects and medico-legal implications of MR-arthrography as well as its various indications are reviewed in this article. There are three possible techniques: indirect MR-arthrography with IV Gadolinium injection, direct MR-arthrography with intra-articular Gadolinium injection and lastly, direct MR-arthrography with intra-articular injection of iodinated contrast media (or saline solution). Indirect MR arthrography cannot be recommended because of insufficient contrast enhancement and the absence of joint filling. Conversely, direct MR-arthrography allows joint expansion which smooths out capsule and ligaments, better delineates articular surfaces and yields a homogeneous high intensity signal of the entire joint. Direct MR-arthrography with iodinated contrast media combines standard arthrography with conventional MRI. Direct MR-arthrography with intra-articular injection of dilute Gadolinium is associated with T1WI, usually of higher quality than T2WI, even though the latter remains part of the protocol. Although, the last two techniques yield higher image quality and are often performed for various articular pathologies, they should not be randomly carried out in the evaluation of joint pathology. However, they should be recommended as the first step in the diagnosis of painful shoulders or hips in young adults and athletes. PMID- 12910171 TI - [Value of US imaging of metacarpophalangeal joints in patients with early rheumatoid arthritis]. AB - Technological advances in the field of ultrasound imaging may have, especially in metacarpophalangeal joints, an impact on decision making in patients with early rheumatoid arthritis. First, the normal anatomy of the metacarpophalangeal joints is briefly reviewed. Then, the authors describe the main ultrasound imaging findings of early RA. The role of ultrasound imaging in the assessment of therapeutic response as well as the benefit of microbubble ultrasound contrast agents are considered. PMID- 12910172 TI - [Scrotal ultrasound. Part II: Cysts, tumors and rare non tumoral processes]. AB - Pathological processes of the scrotum are very numerous. They are composed by few common well known diseases and a large spectrum of rare lesions. The testis may be involved by some systemic diseases. Ultrasound is the main modality for scrotal imaging and complementary to clinical evaluation. When a tumoral process is suspected, surgery is required to confirm the diagnosis and provide histology. The role of MR remains under evaluation. PMID- 12910173 TI - [MRI study before and after extracorporal shock wave therapy in calcifying tendinitis of the shoulder]. AB - PURPOSE: Extracorporal shock wave therapy has recently been proposed with good results (70%) for treatment of persistent painful calcifying tendinitis of the shoulder. The aim of this study was to evaluate the early impact of shock wave therapy on the anatomic structures of the shoulder. MATERIALS AND METHODS: Eight patients (7 women and 1 man, with mean age of 48 years) were prospectively followed up after undergoing shock wave therapy (1500 pulses with 0.28 mJ/mm2 energy). MRI (T1: 500/12 [TR/TE] and STIR: 7200/60/180/180 [TR/TE/TI/alpha]) were obtained 2 hours before and 15 days after the procedure; in addition for 5 of them one more examination was carried out 6 hours after extracorporal shock wave therapy. RESULTS: There was no significant signal change of the humeral bone or rotator cuff, and the calcification size, when seen (6 times), was unchanged at successive MR examinations. One patient had subcutaneous fat signal change (STIR) next to the zone of impact, which resolved 15 days after the extracorporal shock wave therapy. No bursitis or joint effusion was found. CONCLUSION: Shock wave therapy has no early complications or significant impact on the anatomic structures of the shoulder. PMID- 12910174 TI - [Value of thoracic CT in the management of severe hemoptysis]. AB - PURPOSE: To assess the value of thoracic CT in the management of patients with severe hemoptysis. PATIENTS AND METHODS: Between January 1997 and January 2001, 62 patients were investigated for severe hemoptysis (>300 ml/24H). The protocol, performed before angiography and embolization, included bronchial fiberoptic examination (BFE) followed by thoracic spiral CT-angiography. Data recorded at CT and BFE were the presence and location of bleeding, the etiology of hemoptysis and the therapeutic modality. RESULTS: Nine patients with life-threatening hemoptysis directly underwent bronchial embolization. CT was available in the 53 remaining patients. No abnormality was found in 4 patients. CT assessed the presence (n=49) and the location (n=38) of the bleeding. The etiology was determined in 49 patients. BFE was feasible in 38/53 patients. BFE assessed the presence (n=38) and location (n=15) of the bleeding. The etiology was determined in 12 cases of bronchial tumour. The available findings of CT and BFE for the presence and the location of the bleeding were concordant. Comparing fiberoptic examination and thoracic CT, the percentages of localized bleedings (39% and 72%) and demonstrated etiologies (32% and 92%), were significantly different (p<0,005 and p<0.0001 respectively). CONCLUSION: Although retrospective and limited by the small number of cases, our study provides arguments to perform thoracic CT before bronchial fiberoptic examination for the management of severe hemoptysis. PMID- 12910175 TI - [Value of percutaneous core needle biopsy in the investigation of a suspected bone tumor]. AB - PURPOSE: To Determine the value of percutaneous core needle biopsy in the investigation of a suspected bone neoplasm. MATERIAL AND METHODS: We performed a retrospective study of 91 core needle biopsies performed between May 1995 and October 2001. Patients were excluded if they had a known primary carcinoma or if an infection was suspected by clinical, physical or laboratory findings. The results were correlated to the analysis of the surgical piece or, for the 28 patients who did not undergo surgery, to the clinical evolution over more than 12 months. RESULTS: The final diagnosis was metastasis in 29 cases, primary bone tumor in 36 cases and benign lesions in 25 cases. Sensitivity was 92.3% and specificity was 97.4%. For primary malignant bone tumors, results respected histology features and grade in 79.2%. In cases of mistake, because of the correlation of these results to the clinical and radiological context, the treatment of the bony malignant lesions were adapted in 95.8% of cases. Only one major complication was reported in these 91 biopsies. CONCLUSION: First intention core needle biopsy, confronted with radio-clinical context seems to have a place in the evaluation of bone lesions when a tumor is suspected. This technique, less expensive than an open biopsy and with fewer complications, is best performed as part of a multidisciplinary approach with the surgeon's collaboration. PMID- 12910176 TI - [MRI in the diagnosis of Rathke's cleft cyst]. AB - PURPOSE: Rathke's cleft cysts are non neoplastic lesions of the sellar area that seldom are symptomatic. Their incidence has been underestimated before magnetic resonance imaging (MRI). The aim of this work was to assess the value of MRI in the diagnosis and differential diagnosis of Rathke's cleft cyst based on a retrospective review of 12 cases and a review of the literature. PATIENTS AND METHODS: We retrospectively reviewed the MRI features of 12 patients with Rathke's cleft cyst collected over 4.5 years. Patients included ten females and two males (mean age: 39 years). The most common presentation was the association of dysmenorrhea and pituitary dysfunction. MRI examinations were performed using a 1.5 Tesla system, T1 and T2 weighted sequences performed before and after Gd- DTPA injection. Signal intensity, shape, size and location of the lesion were analyzed. RESULTS: In all cases, MRI examination showed a cystic lesion of variable size and signal intensity. The cyst was purely intrasellar in nine cases and a suprasellar extension was noted in three cases. Five patients underwent surgery providing pathological confirmation, four underwent routine follow-up and three were lost of follow-up. CONCLUSION: We conclude that MRI is an efficient tool for diagnosis, allowing appropriate medical decision making. PMID- 12910177 TI - [Spontaneous regression of a focal nodular hyperplasia]. AB - The authors report a case of near complete regression of a focal nodular hyperplasia of the liver over a course of 10 years in a 39 year old woman. This presentation is atypical because of its imaging features (initially typically hypervascular, to eventually become a fibrous scar after undergoing a pseudo angiomatous phase) and because of its near complete spontaneous regression. PMID- 12910178 TI - [Isolated dissection of the superior mesenteric artery]. AB - Isolated spontaneous dissection of the superior mesenteric artery is uncommon. We report a case complicated by arterial rupture and discuss diagnostic imaging work up. Diagnosis is usually made by conventional arteriography. But, dissection can also be diagnosed noninvasively by computed tomography angiography and ultrasound. PMID- 12910179 TI - [Meckel's diverticulum Diagnosed by CT-Enteroclysis]. AB - The authors report the case of a 24 year old woman presenting with acute lower GI hemorrhage caused by a Meckel's diverticulum. Diagnosis was made by CT enteroclysis and confirmed at surgery. This new imaging technique will be briefly described and compared to the other more traditional small bowel imaging techniques. The authors will then present the CT-enteroclysis imaging features of this congenital anomaly. PMID- 12910180 TI - [The piriformis syndrome: a rare cause of non discogenic sciatica. A case report]. AB - Atypical sciatic pain without apparent discogenic origin should raise the possibility of a non-discogenic etiology, especially tumoral but also infectious or inflammatory. We report the case of a young patient presenting with non discogenic S1 radiculopathy due to sciatic nerve involvement in the subpiriformis canal secondary to piriformis muscle inflammation. The authors review technical considerations and results of MRI which is of great value in the evaluation of piriformis muscle pathology. PMID- 12910181 TI - [Liver involvement in HELLP syndrome: CT appearance]. PMID- 12910182 TI - [Fibrolipoma of the terminal filum]. PMID- 12910183 TI - [Recommendations for a QA program for digital mammography]. PMID- 12910184 TI - [Address by the honorary President]. PMID- 12910185 TI - [Five times thirteen...and more. Inaugural address of the President]. PMID- 12910186 TI - [Graduation of the honorary Member: Pr. M. Cloarec]. PMID- 12910187 TI - [Placebo and new therapeutic perspectives for arteriopathy of the lower limbs]. PMID- 12910188 TI - [Clinical and ultrasonographic follow-up after femoro-popliteal venous bypass for lower limb revascularization]. AB - Revascularisation of the lower limbs with vein graft is frequent. Femoro popliteal infrainguinal bypass have good permeability rates at five years. The weak point of autogenous vein bypass grafting is vein graft stenosis, which develops particularly in the first post-operative year. Stenoses are mostly in relation with myo-intimal hyperplasia and are often located at anastomoses. Serial arteriography and echographic studies have shown that the development of intrinsic graft stenosis is found in 20 to 30% of vein conduits in the lower extremity. On the basis of an abundance of clinical data, Duplex ultrasound surveillance of infrainguinal vein graft to detect and repair such lesions before graft occlusion is widely accepted and has shown in many retrospective studies to have a positive influence on lifetime of venous grafts. Critical stenosis (peak systolic velocity (PSV) > 300 cm/s and velocity ration (VR) > 4) should undergo surgical revision as it heralds occlusion of the vein graft in short delays. Intermediate stenosis (peak systolic velocity (PSV) 200-300 cm/s and velocity ratio (VR) 2-4 should be accurately followed as 50 to 70% progress to critical stenosis. Use of echo-doppler monitoring is supported by one randomised study which showed superiority of ultrasound surveillance over clinical surveillance, and by a well described relation between graft thrombosis and an increased amputation rate. In contrast, Duplex ultrasound surveillance for prosthetic grafts is not widely accepted primarily because of the occurrence of rare intrinsic lesions in prosthetic grafts, compared to in vein grafts. Vascular doctors are increasingly concerned with the costs of surveillance program and of surgical interventions. Recent economic appraisal have shown that cost associated with graft maintenance is significant, particularly in the first-year and demands consideration. However revision of a duplex-identified stenosis seems to be less costly than revision after graft thrombosis. Costs of limb amputation and the following expense of postamputation rehabilitation are very high and justify limb salvage-related expense. The results of an ongoing randomised controlled trial (the Vein Graft Trial) comparing duplex ultrasound surveillance against clinical surveillance only will certainly bring more data on the benefits of echographic surveillance. PMID- 12910189 TI - [Hyperplasia of the arterial intima due to smooth muscle cell proliferation. Current data, experimental treatments and perspectives]. AB - Restenosis after vascular surgery using bypasses or endovascular techniques for dilatation or recanalisation remains the major Achilles' heel for these techniques. The progressive decrease of vessel lumen in an anastomose leading to graft failure or after arterial transluminal angioplasty is due to a complex process: intimal hyperplasia. This process can be compared to an hypertrophic healing into the intimal layer, reducing the lumen of the vessel. This process appears shortly after surgery or dilatation, between the 3rd and the 18th month. Mechanisms leading to this process are particularly complex, involving several cells and many regulatory processes still unclear. Smooth muscle cells are the main actor by their ability to proliferate and to secrete matrix into the media layer but stimulation and control of this process appear nevertheless complicated. The present review focuses on the pathophysiology of intimal hyperplasia, on different cells acting and on their regulation. Also, we reviewed the experimental and clinical trials evaluating approaches to the prevention of intimal hyperplasia in arteries. PMID- 12910190 TI - [Complications of endovascular treatment of aneurysm of the infrarenal abdominal aorta: incidence and treatment]. AB - INTRODUCTION: Abdominal aortic aneurysms are the 13th leading cause of death in the United States. Conventional surgical treatment is associated with a low mortality of 1.4-5% and a higher morbidity in high-risk patients. Endovascular aneurysm repair is now performed in patients considered at too high risk for conventional repair. Although the use of endovascular grafts was initially limited, this method is gaining popularity despite the risk of complications including endoleaks, dislocation and graft thrombosis. METHODS: Between June 1997 and June 2000, 28 patients were treated with endovascular stent grafts. 53 patients were treated by open surgical repair. Six patients presenting with rupture were excluded. Endoleaks were detected by arteriogram and computed tomographic scan. The mean aneurysm diameter, with a mean length of 3.2 cm, was 6.3 cm. The mean proximal neck diameter was not greater than 2.4 cm. RESULTS: There were no conversions to open repair. The mean time of the intervention was 103 minutes. Nine patients with type I endoleaks underwent successful endovascular treatment; 2 patients presented a late type I endoleak treated in one case by dilatation. Four patients presented a type II endoleak after 6, 18, 30 and 32 months respectively, treated in two cases by embolization. Finally, erosion of the material was seen in four cases and a migration in one case. A decreased size of the aneurysms was seen in 10 cases, a stabilization in 12 cases, an augmentation of more than 5 mm in one case and a diminution followed by an augmentation in one case. CONCLUSION: Key to success is restrictive patient selection due to morphological criteria and improvements in surgical techniques and equipment to reduce the incidence of specific treatment complications require a long-term follow-up. PMID- 12910191 TI - [Mycotic aneurysm of the extracranial internal carotid artery]. AB - Extracranial internal carotid artery aneurysm is rare. They usually appear as an enlarging pulsatile neck mass with associated tenderness and fever. The diagnosis is difficult and can lead to significant medical morbidity. Treatment of these lesions requires expert surgical management and necessitates an assiduous search for an underlying source. We report a case of a mycotic aneurysm of the extracranial carotid artery in an immunosuppressed patient successfully treated by wide excision and saphenous vein patch angioplasty. PMID- 12910192 TI - [Iatrogenic renal perforation during percutaneous angioplasty]. AB - Percutaneous transluminal angioplasty is recommended as the first choice of therapy for renal artery stenosis. However, morbidity and mortality are significant. Among hemorrhagic complications, renal perforation is unusual. A case is reported, caused by the use of an angled hydrophilic guidewire, in which the diagnosis has been made by computed tomography angiography with typically perirenal hematoma and extravasation of contrast material corresponding to the perforation. This complication has been successfully treated with conservative measures. PMID- 12910193 TI - [Incidence and factors influencing glaucoma after penetrating keratoplasty]. AB - PURPOSE: To determine the incidence and factors influencing glaucoma following penetrating keratoplasty. MATERIAL AND METHOD: We prospectively studied 440 consecutive patients undergoing penetrating keratoplasty over a period of 5 years, with an average follow-up of 2 years. High intraocular pressure was defined as IOP over 20 mmHg measured by Goldmann applanation for at least 1 week or if a patient required topical treatment for at least 1 week. Before surgery, 18.7% of the patients presented with glaucoma or high intraocular pressure; 44.9% received a graft for a bullous keratopathy condition, 25.6% had keratoconus, 31.5% received an anterior chamber lens, and 48.5% were phakic. RESULTS: After surgery, 42.2% of the patients had ocular pressure at 20 mmHg or more and required treatment for high ocular pressure. The increase in intraocular pressure appeared after an average delay of 3.3 +/- 4.7 months. The frequency of the increase in intraocular pressure was 54.3% for the bullous keratopathy patients, 26.6% for the keratoconus patients (p<0.001). At the end of the second year, the graft survival was 82.1% in absence of glaucoma, whereas it was 71.5% in cases of glaucoma after keratoplasty. Between all parameters that we studied, high intraocular pressure before surgery, etiology, the status, and receiver age over 60 years were the main factors influencing high intraocular pressure after keratoplasty. Intraocular pressure was correlated with the decrease in endothelial cell density and was at the origin of graft failure. Intraocular pressure correlated with the Best Spectacle Corrected Visual Acuity (BSCVA) by its effect on endothelial cell density and optic nerve destruction. CONCLUSION: The increase in intraocular pressure is a real and serious complication of keratoplasty. Its physiopathology is very complex. Better knowledge of risk factors can be useful in controlling it. PMID- 12910194 TI - [Long-term functional results of unilateral congenital cataract treatment with early surgery: 20 case studies]. AB - PURPOSE: Evaluation of the long-term functional results of the treatment of unilateral congenital cataract (UCC) surgery before age 1. MATERIAL AND METHODS: Retrospective analysis of 20 consecutive cases of UCC operated on before age 1 and followed by the same medical team during the entire treatment period. Lens implantation adopted when surgically possible (15 cases) and medical treatment of amblyopia are described. Functional results are analyzed with a mean follow-up of 6 years (3.5-11 years) and compared with those of 31 similar cases operated on between 1 and 8 years of age. RESULTS: Four cases had less than 0.1 of final visual acuity, 8 cases remained between 0.1 and 0.4, and 8 cases obtained at least 0.5. Functional results with surgical, refractive, visual, oculomotor, and binocular data of all cases are described. Amblyograms are presented. DISCUSSION: Age at surgery, postoperative delay, type of cataract, and refractive progression were not found to be decisive in final results. We discuss the factors that seem important in the visual outcome and consequences on the sound eye of this heavy amblyopia treatment. In addition to early treatment, the absence of surgical complications and the continuity of patching, the quality of the initial organization of therapeutic strategy, and the continuity of the follow-up seem to be major factors for the success of long-term treatment. Treatment and follow-up should be centralized around a specialized team that will help the local team during the entire length of the treatment. PMID- 12910195 TI - [Efficiency of surgical treatment for chronic macular edema due to branch retinal vein occlusion]. AB - INTRODUCTION: Internal limiting membrane peeling has recently given interesting results in the management of macular edema in diabetic patients, even in the absence of vitreomacular tractions. This study was conducted to evaluate the results of a surgical treatment for macular edema resulting from branch retinal vein occlusion. MATERIAL AND METHODS: A nonrandomized prospective study was conducted between March 2001 and April 2002 on 12 eyes of 12 consecutive patients with a visual acuity of 20/40 or less resulting from branch retinal vein occlusion with macular edema. Internal limiting membrane peeling associated with arteriovenous crossing sheathotomy was performed on six eyes and internal limiting membrane peeling alone was performed on six eyes. Patients were tested with ETDRS visual acuity, fluorescein angiography, automated perimetry, and OCT pre- and postoperatively (at 3 and 6 months). RESULTS: At 6 months, postoperative visual acuity was improved in all patients (mean VA=20/40; range, 20/125-20/20), with an average gain of three lines of vision (or 14 ETDRS points) (p=0.002). Fundus examination and fluorescein angiography results were improved in all patients. At 6 months, on automated field testing, the mean corrected defect improved from 3.4+/-0.9dB to 2.3+/-0.9dB (p=0.008). On OCT, mean foveal thickness decreased from 419+/-57 micro m to 233+/-10 micro m (p=0.02). No difference was noted between simple vitrectomy with internal limiting membrane peeling and arteriovenous crossing sheathotomy in terms of visual acuity (p=0.5), visual field (p=0.2), or foveal thickness (p=0.6) improvement. CONCLUSION: Our findings suggest that internal limiting membrane removal for macular edema may improve the functional prognosis in patients with branch retinal vein occlusion. Adventitial sheathotomy did not yield further functional benefits in these cases. PMID- 12910196 TI - [Radial optic neurotomy for severe central retinal vein occlusion: preliminary results]. AB - PURPOSE: To describe the results of radial optic neurotomy for the treatment of severe central retinal vein occlusion. PATIENTS AND METHODS: Prospective noncomparative single-center study. Analysis of ten eyes of ten consecutive patients whose visual acuity was 0.1 or less. They underwent fluorescein angiography, visual field testing by automated perimetry, and macular thickness analysis by optical coherence tomography preoperatively at 3 months and at 6 months postoperatively. RESULTS: Mean visual acuity on an ETDRS chart increased from 30+/-12 points preoperatively to 42+/-15 points at the 3-month visit, (p=0.03), and mean macular thickness decreased from 580+/-150 micro m to 361+/-52 micro m (p=0.04). All patients had clinical improvement as determined by fundus examination and fluorescein angiography. An improvement in the central visual field was observed in all eyes. Mean visual acuity of the five patients followed up for 6 months was 52.8+/-20 points. No visual loss was observed. None of the patients underwent laser photocoagulation or has presented with neovascularization so far. Optociliary veins developed in three eyes and a retinochoroidal anastomosis within the disk incision was observed in two eyes. CONCLUSION: These preliminary results are encouraging when compared to the reported natural progression of severe central retinal vein occlusion. A bypass of the site of occlusion is a possible mechanism for radial optic neurotomy. A randomized study should be conducted to assess the efficacy of radial optic neurotomy and determine the best candidates for surgery. PMID- 12910197 TI - [Calculating the mean visual acuity and the change in visual acuity with a decimal acuity chart]. AB - The decimal visual acuity chart is not easy to use for statistical analysis and requires a transformation into LogMAR units (decimal logarithm of the Minimum Angle Resolution). In contrast to the decimal chart, the logarithmic chart has an arithmetic progression and a constant interval between lines. The LogMAR chart makes statistical analysis of visual acuity easy. Change in visual acuity is calculated directly by subtracting LogMAR data, while the average visual acuity is obtained with the arithmetic mean value of the LogMAR data. The mean acuity expressed in LogMAR units can be transformed into a decimal chart for a more comprehensive result. To calculate the average visual acuity directly from the decimal data, the geometric mean value must be used instead of the arithmetic mean value. PMID- 12910198 TI - [Is cheating with automatic perimetry possible?]. AB - INTRODUCTION: Automatic perimetry examination requires the patient's cooperation, but the programs used check response validity. This study evaluated the possibility of patient cheating on this examination. MATERIALS AND METHOD: This study investigated 27 visual fields of automatic perimetry with patients who had no ophthalmological history. Each subject had to reproduce a visual field loss that was observed for 5 minutes (two diffuse losses, five fields reflecting hemianopsia, four reflecting quadranopsia, two tubular losses, five nasal steps, nine absolute and arciform losses). The 24-2 Fastpac of the Humphrey field analyzer and the G2 of the Octopus perimeter were used for this study. RESULTS: Simple deficits were easy to reproduce, but visual field defects are often deeper and simulated test results tend to be too perfect. On the other hand, complex defects such as arciform scotoma or nasal steps were more difficult to reproduce. Visual field defect simulations were exaggerated beyond what was reasonable. CONCLUSION: This study reports on whether it is possible to cheat with automatic perimetry. The different parameters analyzed by computer programs are not able to detect cheating patients. PMID- 12910199 TI - [Phototoxic drugs and age-related maculopathy]. AB - OBJECTIVES: To disclose a correlation between age-related macular degeneration (AMD) and long-lasting treatments with potentially phototoxic drugs. DESIGN: Case control study. METHODS: The case group was made up of 90 randomized patients, 65 years old or older, with documented AMD, examined in 1998 and 1999. The M/F sex ratio was 0.54, and mean age 79.9 years. The control group was randomized with age- and sex-matched subjects. In both groups, we noted long-lasting oral treatment for hypertension, diabetes mellitus, epilepsy, hyperuricemia, and psychiatric pathologies. All the study group patients underwent a complete ophthalmological examination for visual acuity, contrast sensitivity, biomicroscopy, fundus color and red-free photographs, fluorescein and ICG angiographies. RESULTS: When the study group was compared with the control group, neovascular AMD was positively associated with thiazide diuretics (p<0.001). When the study group was analyzed alone, intake of one or several antihypertensive drugs was positively associated with occult neovascularization and mild to moderate visual loss (p=0.009). Only thiazide diuretic intake was positively associated with severe bilateral loss of vision (p=0.041) and extensive choroidal neovascularization with major subretinal hemorrhages (p=0.022). CONCLUSIONS: These findings suggest that severe neovascular AMD is associated with long-term thiazide diuretic treatment. PMID- 12910200 TI - [Feeder vessel treatment of choroidal neovascularization in age-related macular degeneration]. AB - INTRODUCTION: To report on the results of feeder vessel treatment for choroidal neovascularization in age-related macular degeneration (AMD). MATERIAL AND METHODS: Retrospective study of 40 patients presenting subfoveal classic choroidal neovascularization (CCNV) and/or occult choroidal neovascularization (OCNV) with or without fibrovascular pigment epithelium detachment (PED), based on fluorescein angiography (FA) and indocyanine green angiography (ICG). Visual acuity was measured using Snellen lines and/or ETDRS. The feeder vessels were visualized with high-speed ICG angiography (confocal scanning laser ophthalmoscope, Heidelberg HRA, Heidelberg, Germany). Feeder vessels were identified as very thin vessels with early filling and rapid emptying. Feeder vessel closure was achieved by laser photocoagulation treatment with either minipulse infrared laser therapy (810 nm), microburst, or yellow laser (568 nm). Post-treatment high-speed ICG angiography follow-up was recommended to the patient immediately or the day after the treatment. The number of treatments and location of the feeder vessels in relation to the macula as well as FA, ICG, and visual acuity results were assessed. If necessary, other treatment techniques such as photodynamic therapy (PDT) or transpupillary thermotherapy (TTT) were offered to the patient when leakage was still present. RESULTS: The patients were followed for an average of 6.2 months (2-9 months). Twenty-six women and 14 men (mean age, 72 years; range, 51-95) were included. Eight patients demonstrated classic neovascularization, 32 presented occult neovascularization, including 17 cases of vascularized PED and 15 retrofoveal plaques. The average number of treatments was 1.8 (range, 1-4 treatments). Location of feeder vessels were as follows: nine superior-macular, 18 inferior-macular, six nasal-macular and 11 temporal-macular. Visual acuity improved two lines or more in nine patients (22.5%), decreased 2 lines or less in five patients (12.5%) and was stable in 26 patients (65%). After feeder vessel treatment, seven patients showed improvement of the anatomical aspect of the AMD lesions, with a reduction in leakage. Except for one subretinal hemorrhage, no complications after feeder vessel treatment were observed. CONCLUSION: Feeder vessel treatment is a technique requiring high speed ICG angiography to detect the feeder vessels located at a distance from the subfoveal OCNV, CCNV, and PED. Although the final visual acuity result was for the most part stabilization, the technique appeared to be safe and reproducible. In patients who were not eligible for other therapies such as PED, feeder vessel treatment seemed to be a reasonable alternative. PMID- 12910201 TI - [Uveitis in children: about 20 cases]. AB - INTRODUCTION: Uveitis in children is rare: 3%-8% of all cases. It requires the same diagnostic and etiological processes as uveitis in adults, with additional difficulties at examination and a sometimes insidious progression. MATERIAL AND METHODS: Retrospective study of 20 cases of childhood uveitis from 1995 to 2000. All patients received an ophthalmologic examination and an etiological search, with specific and/or symptomatic treatment and follow-up lasting from 6 months to 5 years. RESULTS: The 20 children presenting uveitis were 4-16 years old, with etiologies as follows: 8 cases of Behcet's disease, 2 cases of Vogt-Koyanagi Harada's disease, 1 case of sarcoidosis, 1 case of uveitis associated with coeliac disease, 1 case of toxoplasmosis, 1 case of sympathetic ophthalmia, 1 case of uveitis with streptococcal infection, 3 cases of ocular toxocarosis, and 2 cases with unknown etiology. Treatment based on the etiological findings was started in the cases of toxoplasmosis and uveitis from streptococcal infection. The others were treated with high- and then digressive-doses of corticosteroids. ANALYSIS: We have noted the high incidence of Behcet's disease in our series. Progression was marked by frequent recurrence for one case of Vogt-Koyanagi Harada's syndrome and one case of toxoplasmosis. This study also revealed a few cases of complicated cataract. DISCUSSION: Clinical characteristics, diagnosis, and treatment of uveitis in children are discussed. PMID- 12910202 TI - [Choroidal granuloma revealing tuberculosis. A case report]. AB - BACKGROUND: The incidence of tuberculosis is still high in developing countries and a steady increase in new cases has been observed in industrial countries within the past decade. Among other reasons, the growing number of immunodeficient patients and migration from developing to industrial countries are both contributing factors. OBSERVATION: A 70-year-old female patient presented with a marked decrease in visual acuity of the left eye. There was no history of tuberculosis. Funduscopy of the left eye revealed a choroidal tumor with multiple choroidal lesions, mostly located at the posterior pole, and adjacent serous retinal detachment. Ultrasonography showed a solid elevated mass; magnetic resonance imaging showed the tumor with posterior extension. Because of blindness in this eye and the presence of posterior extension, the eye was subsequently enucleated. Intraoperative observation found caseum necrosis through scleral breaking. The histological study confirmed choroidal tuberculoma. The patient developed pulmonary tuberculosis 15 days after surgery. The outcome was favorable after antibiotic treatment. DISCUSSION: Confirming the diagnosis of tuberculous uveitis is often difficult. The differential diagnosis includes other granulomatous ocular inflammations. The detection of Mycobacterium tuberculosis and the clinical course make this diagnosis the most likely one. CONCLUSION: Choroidal granuloma is a rare and atypical location of tuberculosis. This case showed the diagnostic difficulties when there is no history of tuberculosis. PMID- 12910203 TI - [Dermoid cysts: epidemiological, clinical and pathological studies]. AB - We retrospectively analyzed 31 cases of dermoid cysts surgically treated between January 1992 and October 2000. Mean patient age was 18.9 years. The preseptal localization predominated, with 29 cases. Two cases of intraorbital localization required orbitotomy. The surgical result was excellent. PMID- 12910204 TI - [Acute bilateral anterior uveitis with hypopyon revealing creeping angle-closure glaucoma]. AB - We report the case of a 35-year-old Asian woman presenting with bilateral anterior uveitis with hypopyon associated with high intraocular pressure. In her past history, she described intermittent angle-closure episodes. Anterior chamber examination and ultrasound biomicroscopy revealed a creeping angle-closure glaucoma. This clinical entity can affect Asians. The angle is gradually shortened with an iris bombe that brings the peripheral iris close to the external angle wall. The acute episodes are often poorly symptomatic. An early diagnosis may prevent glaucoma damages. PMID- 12910205 TI - [Lymphoma revealed by isolated obliquus inferior muscle involvement in exophthalmia]. AB - Orbital lymphomas account for 4% of nontraumatic orbital disorders. We report the case of a 54-year-old patient who presented with binocular diplopia associated with right exophthalmia with an isolated obliquus inferior localization. This lymphomatous localization was unexpected, as lymphomas are described more often in the rectus superior and inferior muscles. Pathological examination found a low grade non-Hodgkin's MALT lymphoma. This case is the basis of a review of the various diagnoses and the necessity of a biopsy prior to surgical tumor ablation that may damage an oculomotor muscle. Treatment should be medical: first intention surgery would have abolished obliquus inferior function. PMID- 12910206 TI - [Compressive optic neuropathy caused by fibrous dysplasia]. AB - We report a case of compressive optic neuropathy caused by fibrous dysplasia in a 28-year-old woman. The patient had no significant medical history. Her best corrected visual acuity was 20/20 in the right eye and 20/400 in the left eye. There was an afferent pupillary defect in the left eye. Slit-lamp examination was unremarkable. Funduscopy showed a normal optic disc bilaterally and congenital hypertrophy of the retinal pigment epithelium in the right eye. Systemic evaluation disclosed facial asymmetry and mucocutaneous lentiginosis involving the face and the limbs. Goldmann visual field testing showed a cecocentral scotoma in the left eye. Imaging studies demonstrated extensive changes of craniofacial fibrous dysplasia involving the sphenoid bone, with compression of the left optic nerve by a cystic structure. Results of gastrointestinal fibroscopy were unremarkable. The patient was given systemic steroids. After 4 days of treatment, her visual acuity had improved to 20/40, with resolution of the afferent pupillary defect and visual field improvement. Debulking of the tumor was recommended, but the patient declined the procedure. She has remained stable over 13 months of follow-up. Compressive optic neuropathy should be considered as a leading cause of visual loss in patients with craniofacial fibrous dysplasia. Early diagnosis, close follow-up, and appropriate management are mandatory to improve or maintain the visual function in such patients. PMID- 12910208 TI - [Pulmonary hypertension revealed by bilateral venous occlusion associated with ocular hypertension]. PMID- 12910207 TI - [Immunosuppression in corneal transplantation]. AB - Rejection is the leading cause of corneal graft failure, induced by loss of the so-called eye immune privilege. Prevention of graft rejection with immunosuppressive therapy is then necessary. Topical corticosteroids are currently the gold standard, and steroids are the only treatment for acute rejection episodes. Steroids are nonspecific immunosuppressive agents, and they can induce glaucoma, cataract, infections, and epithelial defects. Cyclosporin has a specific effect, because it inhibits interleukin-2 transcription and, consequently, the specific activation of T lymphocytes. When cyclosporin is given orally, it effectively prevents graft rejection in high-risk recipients, but it may induce severe side effects (i.e., systemic hypertension, kidney deficiency, and malignant tumor induction). When cyclosporin is given topically, it can effectively replace steroids in case of dexamethasone-induced glaucoma and graft infection, but it can also induce serious corneal epithelial defects. Cyclosporin is not a treatment for acute rejection episodes. Mycophenolate mofetil and FK 506 are promising drugs, but currently they cannot be used routinely to prevent corneal graft rejection. PMID- 12910209 TI - [Optic disk neovascularization in fungal endophthalmitis]. AB - A case of optic disk neovascularization is reported in a 30-year-old female heroin addict with suspected fungal endophthalmitis treated with an intravitreal injection of amphotericin B and fluconazole (200mg/day). Membrane healing was characterized by densification and contraction leading to tractional retinal detachment; excision by vitrectomy was required. Electron microscopy analysis of the membrane removed the tenth month revealed a poorly vascularized fibrous tissue infiltrated by chronic inflammatory cells. Visual improvement was poor owing to a postoperative macular disorder and a full-thickness retinal scar. PMID- 12910210 TI - [Echography for inguinal hernia: will it ever have a factual basis?]. PMID- 12910212 TI - [Management of anastomotic fistula following excision of rectal cancer]. AB - The risk of anastomotic leak after resection of cancers of the mid or low rectum with mesorectal excision is about 10%--the lower the colo-rectal or colo-anal anastomosis, the higher the risk of leak. If the fistula is asymtomatic and the leak is walled off, it is best to defer the closure of the diverting ileostomy for 2-3 months and to proceed only when a radiologic contrast study shows the fistula to have disappeared. More commonly, the anastomotic fistula presents as a pelvic abscess. It is simple and logical to drain the abscess into the digestive tube by enlarging the orifice of the fistula; this can usually be done with a brief general anesthetic. Less commonly, the abscess may present at some distance from the anastomotic leak; this calls for percutaneous drainage. If abscess drainage fails, if pelvic sepsis persists, or if the leak presents from the start as generalized peritonitis, laparotomy is called for in order to lavage the abscess cavity, place effective drains, and perform, if necessary, a diverting stoma upstream. Two strategies are possible: 1) drain placement at the leak site with upstream loop diverting stoma, or 2) takedown of the anastomosis, closure of the distal stump as a Hartmann pouch, and proximal end colostomy in the left lower quadrant. In the first instance, one must be sure the fistula has healed before stoma closure. In the second, the problem is to obtain (at a second stage) sufficient length of well-vascularized proximal colon to make an anastomosis to a short Hartmann pouch or to the anus in a pelvis scarred and inflamed by infection and radiation. A Soave procedure may allow an anastomosis with less risk to peri rectal innervation and with less blood loss. Two maneuvers which may help to gain length are the Toupet technique for freeing the transverse mesocolon or the Deloyer technique of mobilizing the hepatic flexure. In the face of post operative pelvic sepsis, an early intervention adapted to the circumstances will increase the chances of healing and reestablishment of intestinal continuity, and may avoid multiple complex interventions with poor functional results including incontinence, urgency, and difficult evacuation. PMID- 12910211 TI - [Leiomyosarcoma of the inferior vena cava]. AB - Leiomyosarcoma of the inferior vena cava is a rare tumor of mesenchymal origin most commonly found in women. Clinical signs are non-specific. Imagery with ultrasound, CT, or MRI may strongly suggest the diagnosis, but it can only be confirmed by histologic examination of tissue obtained pre or intra-operatively. The tumor is slow growing but nonetheless carries a bad prognosis; it may grow to a large size before directly invading adjacent structures. Systemic spread is a late occurrence. Radical surgical resection is the only treatment which offers any hope for prolonged survival. Standard vascular surgical techniques are usually sufficient. Progress in the techniques of hepatectomy and liver transplantation have allowed the experienced surgeon to undertake the removal of retrohepatic lesions once considered unresectable. High-lying lesions adjacent to the hepatic veins or with thrombus extending into the proximal vena cava may require extracorporeal circulation with or without profound hypothermic circulatory arrest. The efficacy of chemotherapy, whether pre-operative for inaccessible tumors or post-operative for incompletely resected or recurrent tumor, is poorly defined and very limited. PMID- 12910213 TI - [Laparoscopic treatment of rectal prolapse with associated sigmoid colectomy]. PMID- 12910214 TI - [Treatment of cardiac wound]. PMID- 12910215 TI - [The pneumopertitoneum in laparoscopy: Part 3. Directed open insertion of a first trocar]. PMID- 12910216 TI - [Gallstone ileus]. PMID- 12910217 TI - [Where does laparoscopy fit in the treatment of inguinal hernia in 2003?]. AB - Meta-analysis of randomized studies has clearly shown that prosthetic repair of inguinal hernias decreases the risk of hernia recurrence when compared with herniorraphy without prosthesis; but the optimal route for insertion of the prosthetic patch (laparoscopic versus open inguinal approach) remains in dispute. Meta-analysis of randomized studies comparing laparoscopic with open prosthetic hernia repair suggest that laparoscopy is associated with less post-operative pain (both early and late), a quicker recovery, and earlier return to work. Yet this is at the price of longer operative time and an incidence of rare but potentially severe complications. On the basis of these randomized studies, the ANAES in France and the NICE in England have put forth recommendations which accept the indication for laparoscopic repair in recurrent and bilateral hernias, if done by surgeons experienced in laparoscopic technique. For unilateral hernia in adults, laparoscopic repair has shown no proof of superiority over open prosthetic repair in terms of mortality, morbidity, or recurrence rate. The principal advantage of the laparoscopic approach seems to be improved patient comfort; its disadvantage is higher cost and technical difficulty with a prolonged learning curve. The excess costs of the laparoscopic approach may be compensated by an earlier return to work. At present, the laparoscopic repair of hernias finds its clinical niche in patients with bilateral or recurrent hernias or in patients with unilateral hernia who desire a minimal period of postoperative disability. PMID- 12910218 TI - [Cancer of the gastric antrum]. PMID- 12910220 TI - [Operating tools useful for hepatitis C poles and care networks]. PMID- 12910221 TI - [Role of transjugular intrahepatic portosystemic stents (TIPS) in the prevention of recurrent bleeding due to portal hypertension]. PMID- 12910222 TI - [Treatment of chronic hepatitis C with interferon alpha and ribavirin. Results in "real life"]. AB - OBJECTIVES: The applicability of the results of controlled, randomised, therapeutic trials in the general population is not well known. The aim of this prospective study was to assess the results of the treatment of chronic hepatitis C with interferon alpha and ribavirin in "real life." METHODS: In an independent observatory, clinicians from 31 hepatogastroenterology units in French general hospitals registered patients treated with interferon alpha and ribavirin outside clinical trials. Results were compared with those of the best arm (twelve months of bitherapy) of the European-Canadian trial published in 1998 by Poynard et al. RESULTS: 262 naive patients were included. Patients were significantly older, had more severe liver fibrosis, with cirrhosis in 17 percent, but were comparable for sex, contamination mode and viral genotype with those of the Poynard et al. trial. Sustained virological response rate (37%, IC 95%: 31-43%) was not significantly lower (difference 6%, IC 95%: - 18% to + 6%) than in the Poynard et al. trial. Results for response factors were similar. Comorbid conditions which would have prevented our inclusion of patients in the Poynard et al. trial were present in 46% and had an independent negative effect on sustained virological response rate. Side-effects were similar. CONCLUSIONS: The efficacy of the combination of interferon alpha and ribavirin remains good in real life, but depends on the characteristics of the treated population. The results of trials should be presented more cautiously to general population. PMID- 12910223 TI - [Hepatitis C virus genotype 4: epidemiology and treatment]. PMID- 12910224 TI - [Creation of a center for OMGE post-graduate education in gastroenterology in Rabat, Morocco. A great opportunity for French-speaking gastroenterologists in Africa]. PMID- 12910225 TI - [Digestive cancers: the new frontier of hepatogastroenterology]. PMID- 12910226 TI - [Endoscopic therapy of benign colonic post-operative strictures: report on 27 cases]. AB - Benign postoperative anastomotic strictures are frequent. OBJECTIVES: To evaluate the results of endoscopic dilatations and to propose an alternative treatment to surgery. METHODS: Between 1994 and 2001, 27 patients (16 female, 11 male, median 70 years old) with colonic anastomotic strictures were treated with pneumatic or mechanical dilatation. Eleven patients (40.7%) had been operated on for colon carcinoma, 2 for colorectal adenoma and 14 (51.8%) for complicated diverticular disease. A left colectomy was performed in 12 patients and a sigmoid colectomy in 15. Anastomoses were stapled in 25 cases. RESULTS: Twenty one patients were symptomatic (78%). The median time to diagnosis after surgery was 3 months (range: 21 days-4 years). Dilatation was possible in 26 cases (96.3%). The total number of dilatation sessions was 51 and the median number of sessions by patient was 2 (range: 1-4). Only one session was done in 11 patients (40.7%). There was one case of bowel perforation (1.9%) with the guide wire. In 21 cases (77.7%), the patients were no longer symptomatic and failure was observed in 5 cases. Surgery was necessary for 3 patients, with stenosis recurrence in 1. Three patients were treated with a self expanding metal stent. CONCLUSION: Endoscopic dilatation is a simple and efficient method. Therefore, it might be considered as the first line approach for benign anastomotic strictures. Self expanding metal stent could be an alternative to surgery, in case of failure. PMID- 12910227 TI - Enteral nutrition during acute pancreatitis: feasibility study of a self propeeling spiral distal end jejunal tube. AB - OBJECTIVE: The aim of our study was to evaluate the feasibility of enteral jejunal nutrition for acute pancreatitis using a self-propelling spiral distal end jejunal tube. METHODS: Sixteen consecutive patients with acute pancreatitis in whom Flocare tubes were placed for enteral nutrition were included in this open prospective study. All of them had pancreatic and/or peripancreatic necrosis (Balthazar >=D). The median computed topography index was 5 (range 3-10) and the median Ranson score was 2 (range 0-5). The nasoenteric Flocare tube (spiral distal end) was inserted in the stomach at the bedside. Self progression into the jejunum was assessed by X-ray at 1, 7 and 12 hours and then every 24 hours for 4 days. The rate of successful tube self-placement in the jejunum and the time to successful placement were noted. RESULTS: Insertion was successful in 12 of 16 patients (75%). Treitz's ligament was reached in a median of 12 hours (range 1-96 hours). For the remaining patients, the tube was successfully repositioned under fluoroscopic guidance in 2 and withdrawn in 2, one for oral renutrition and one to change to a weighted jejunal tube. No tube dysfunction or recurrence of pancreatitis occurred during the entire period of enteral nutrition. CONCLUSIONS: This study suggests that the nasoenteric Flocare tube can be used effectively and safely in early enteral jejunal nutrition for severe acute pancreatitis, without endoscopic or radiological manipulation. PMID- 12910228 TI - [Inflammatory bowel disease and pregnancy]. PMID- 12910229 TI - [Indications for stool tests in adults]. PMID- 12910230 TI - [Portal vein thrombosis associated with factor V Leiden mutation in a woman who underwent exchange transfusion at birth]. AB - We report the case of a 18-year-old woman with portal vein thrombosis and chronic hepatitis C virus. Portal vein thrombosis was diagnosed by chance on ultrasound examination during initial hepatitis C virus-positive patient screening. The patient interview revealed a history including exchange transfusion at birth, followed by necrotising ulcerocolitis and septicemia. The investigation of general factors favoring thrombosis showed acquired protein S deficiency and heterozygous factor V Leiden mutation. This case demonstrates the value of systematic investigations for general thrombophilic factors in cases of portal vein thrombosis even when a local cause is found. PMID- 12910231 TI - [Hematochezia due to jejunal diverticula and treated by arterial embolization]. AB - Digestive bleeding is a rare complication of jejunal diverticulas whose diagnosis and treatment are difficult. We report a case of hematochezia whose origin was not determined by endoscopic investigations. Coelio-mesenteric arteriography showed a leakage of contrast medium from a jejunal arterial branch and allowed a successful embolization of the arterial branches feeding the bleeding point. PMID- 12910232 TI - [Presence of microcysts within a cystic tumor of the pancreas does not rule out malignancy: report of two cases]. AB - Presence of microcysts within a cystic tumor of the pancreas is usually considered as specific of serous cystadenoma and leads to conservative management and surveillance of asymptomatic patients. We report two cases of malignant cystic tumors of the pancreas with a microcystic component detected by endosonography. We conclude that presence of microcysts detected by endosonography in cystic tumors of the pancreas does not rule out malignancy. PMID- 12910233 TI - [Malacoplakia of the biliary tract and duodenum. Case report]. PMID- 12910234 TI - [Reappearance of syphilis and an old cause of hepatitis]. PMID- 12910235 TI - [Intragastric then intraesophageal migration of an endo-esophageal prosthesis]. PMID- 12910236 TI - [Weber Christian syndrome due to pancreaticomesenteric fistula secondary to acute alcoholic pancreatitis: treatment with pancreatic sphincterotomy alone]. PMID- 12910237 TI - [Endoscopic treatment of severe ileal bleeding in Crohn's disease]. PMID- 12910238 TI - [Immunoallergic shock to oxaliplatin: a new case]. PMID- 12910239 TI - [Gene therapy for liver fibrosis using genes that code for protease matrices]. PMID- 12910240 TI - Down-regulation of RPE65 protein expression and promoter activity by retinoic acid. AB - PURPOSE: RPE65 is critical for the normal formation of 11-cis retinal and thus photoreceptor function. Opsin expressed in HEK293 cells has been reported to form rhodopsin on the addition of all-trans retinol, indicating that the machinery for retinoid isomerization is present. RPE65 has been previously identified in HEK293 cells at both the RNA and protein levels. To further understand retinoid metabolism in these cells and the control of RPE65 expression, HEK293 cells were used as a model to determine if retinoic acid (RA) affects RPE65 promoter activity. METHODS: RPE65 levels were determined by Western blots. RA regulation of RPE65 promoter activity was monitored using the luciferase reporter assay after transient transfection of HEK293 cells with the RPE65 promoter. Deletion and truncation promoter mutants were assessed for activity. RESULTS: RA down regulates RPE65 protein expression and promoter activity. The RA receptors (RARs), RARalpha, -beta, and -gamma, and the retinoid X receptors (RXRs), RXRalpha, -beta, and -gamma, were all identified in these cells and shown to mediate the regulation of RPE65 mRNA expression. After deletion of the AP1, AP4 or NF1 transcription factor binding sites, the RA down-regulation was decreased, but the decrease was not associated with a single transcription factor. The truncation promoter constructs P60, P153 and P257 showed increases in promoter activity, indicating an inhibitory element had been removed, and the down regulatory effect of RA was decreased. CONCLUSIONS: The down-regulation of RPE65 by RA is occurring at the transcription level. Multiple elements in the RPE65 promoter may contribute to this regulation. PMID- 12910241 TI - Acoustic factors govern developmental sharpening of spatial tuning in the auditory cortex. AB - Auditory localization relies on the detection and interpretation of acoustic cues that change in value as the head and external ears grow. Here we show that the maturation of these structures is an important determinant for the development of spatial selectivity in the ferret auditory cortex. Spatial response fields (SRFs) of high-frequency cortical neurons recorded at postnatal days (P) 33-39 were broader, and transmitted less information about stimulus direction, than in older ferrets. They also exhibited slightly broader frequency tuning than neurons recorded in adult animals. However, when infant neurons were stimulated through virtual ears of adults, SRFs sharpened significantly and the amount of transmitted information increased. This improvement was predicted by a model that generates SRF shape from the localization cue values and the neurons' binaural spectrotemporal response properties. The maturation of spatial response characteristics in auditory cortex therefore seems to be limited by peripheral rather than by central factors. PMID- 12910242 TI - Retrograde regulation of synaptic vesicle endocytosis and recycling. AB - Sustained release of neurotransmitter depends upon the recycling of synaptic vesicles. Until now, it has been assumed that vesicle recycling is regulated by signals from the presynaptic bouton alone, but results from rat hippocampal neurons reported here indicate that this need not be the case. Fluorescence imaging and pharmacological analysis show that a nitric oxide (NO) signal generated postsynaptically can regulate endocytosis and at least one later step in synaptic vesicle recycling. The proposed retrograde pathway involves an NMDA receptor (NMDAR)-dependent postsynaptic production of NO, diffusion of NO to a presynaptic site, and a cGMP-dependent increase in presynaptic phosphatidylinositol 4,5-biphosphate (PIP2). These results indicate that the regulation of synaptic vesicle recycling may integrate a much broader range of neural activity signals than previously recognized, including postsynaptic depolarization and the activation of NMDARs at both immediate and nearby postsynaptic active zones. PMID- 12910243 TI - Isolation of drugs active against mammalian prions using a yeast-based screening assay. AB - We have developed a rapid, yeast-based, two-step assay to screen for antiprion drugs. The method allowed us to identify several compounds effective against budding yeast prions responsible for the [PSI+] and [URE3] phenotypes. These inhibitors include the kastellpaolitines, a new class of compounds, and two previously known molecules, phenanthridine and 6-aminophenanthridine. Two potent promoters of mammalian prion clearance in vitro, quinacrine and chlorpromazine, which share structural similarities with the kastellpaolitines, were also active in the assay. The compounds isolated here were also active in promoting mammalian prion clearance. These results validate the present method as an efficient high throughput screening approach to identify new prion inhibitors and furthermore suggest that biochemical pathways controlling prion formation and/or maintenance are conserved from yeast to humans. PMID- 12910244 TI - Multicolor in vitro translation. AB - In vitro translation is a widely used tool for both analytical and preparative purposes. For analytical purposes, small amounts of proteins are synthesized and visualized by detection of labeled amino acids incorporated during translation. The original strategy of incorporating radioactively labeled amino acids, such as [35S]methionine or [14C]leucine, has been superseded by the addition of antigenic tags or the incorporation of biotin-labeled or BODIPY-FL-labeled amino acids. Such nonradioactive tags are easier to visualize after translation and do not pose a radiation hazard. Among the nonradioactive tags, BODIPY-FL-lysine offers the advantage that proteins that have incorporated this amino acid can be directly visualized after gel electrophoresis. We show here that multiple fluorophores introduced into proteins can considerably extend their usefulness, particularly for the comparison of in vitro-translated proteins from related sources. This technology can be applied in various situations, including the simplified detection of rare truncating mutations in clinical samples from cancer patients. PMID- 12910245 TI - High-level generation of polyclonal antibodies by genetic immunization. AB - Antibodies are important tools for investigating the proteome, but current methods for producing them have become a rate-limiting step. A primary obstacle in most methods for generating antibodies or antibody-like molecules is the requirement for at least microgram quantities of purified protein. We have developed a technology for producing antibodies using genetic immunization. Genetic immunization-based antibody production offers several advantages, including high throughput and high specificity. Moreover, antibodies produced from genetically immunized animals are more likely to recognize the native protein. Here we show that a genetic immunization-based system can be used to efficiently raise useful antibodies to a wide range of antigens. We accomplished this by linking the antigen gene to various elements that enhance antigenicity and by codelivering plasmids encoding genetic adjuvants. Our system, which was tested by immunizing mice with >130 antigens, has shown a final success rate of 84%. PMID- 12910246 TI - Protein microarrays guide tolerizing DNA vaccine treatment of autoimmune encephalomyelitis. AB - The diversity of autoimmune responses poses a formidable challenge to the development of antigen-specific tolerizing therapy. We developed 'myelin proteome' microarrays to profile the evolution of autoantibody responses in experimental autoimmune encephalomyelitis (EAE), a model for multiple sclerosis (MS). Increased diversity of autoantibody responses in acute EAE predicted a more severe clinical course. Chronic EAE was associated with previously undescribed extensive intra- and intermolecular epitope spreading of autoreactive B-cell responses. Array analysis of autoantigens targeted in acute EAE was used to guide the choice of autoantigen cDNAs to be incorporated into expression plasmids so as to generate tolerizing vaccines. Tolerizing DNA vaccines encoding a greater number of array-determined myelin targets proved superior in treating established EAE and reduced epitope spreading of autoreactive B-cell responses. Proteomic monitoring of autoantibody responses provides a useful approach to monitor autoimmune disease and to develop and tailor disease- and patient-specific tolerizing DNA vaccines. PMID- 12910247 TI - Molecular viral oncology of hepatocellular carcinoma. AB - Hepatocellular carcinoma (HCC) is the fifth most common cancer, but the third leading cause of cancer death, in the world, with more than 500,000 fatalities annually. The major etiology of HCC/liver cancer in people is hepatitis B virus (HBV), followed by hepatitis C virus infection (HCV), although nonviral causes also play a role in a minority of cases. Recent molecular studies confirm what was suspected: that HCC tissue from different individuals have many phenotypic differences. However, there are clearly features that unify HCC occurring in a background of viral hepatitis B and C. HCC due to HBV and HCV may be an indirect result of enhanced hepatocyte turnover that occurs in an effort to replace infected cells that have been immunologically attacked. Viral functions may also play a more direct role in mediating oncogenesis. This review considers the molecular and cellular mechanisms involved in primary hepatocellular carcinoma, using a viral perspective. PMID- 12910248 TI - Epstein-Barr virus and oncogenesis: from latent genes to tumours. AB - Epstein-Barr virus (EBV) is a ubiquitous human herpesvirus associated with the development of both lymphoid and epithelial tumours. As a common virus infection, EBV appears to have evolved to exploit the process of B cell development to persist as a life-long asymptomatic infection. However, the virus can contribute to oncogenesis as evidenced by its frequent detection in certain tumours, namely Burkitt's lymphoma (BL), post-transplant B cell lymphomas, Hodgkin's disease (HD) and nasopharyngeal carcinoma (NPC), and by its unique ability to efficiently transform resting B cells in vitro into permanently growing lymphoblastoid cell lines (LCLs). These transforming effects are associated with the restricted expression of EBV genes such that only a subset of so-called latent virus proteins are expressed in virus infected tumours and in LCLs. Distinct forms of EBV latency are manifest in the different tumours and these appear to be a vestige of the pattern of latent gene expression used by the virus during the establishment of persistent infection within the B cell pool. This review summarises our current knowledge of EBV latent gene function and how this relates to the role of the virus in the aetiology of different tumours. PMID- 12910250 TI - Human T-cell leukemia virus type I and adult T-cell leukemia. AB - Human T-cell leukemia virus type I (HTLV-I) causes adult T-cell leukemia (ATL) in about 5% of carriers after a long latent period. After its infection, HTLV-I promotes the clonal proliferation of HTLV-I infected cells in vivo by actions of encoded viral proteins, including Tax. However, leukemic cells frequently lack the expression of Tax by the genetic and epigenetic changes of HTLV-I provirus, suggesting that Tax is not always necessary after transformation. Alternatively, ATL cells without Tax protein could escape from the host immune system since Tax is the major target of cytotoxic lymphocytes. During the latent period, alterations of host genome accumulate, finally leading to onset of ATL. PMID- 12910249 TI - Virally targeted therapies for EBV-associated malignancies. AB - In Epstein-Barr virus (EBV)-positive lymphomas, the presence of the EBV genome in virtually all tumor cells, but very few normal cells, suggests that novel, EBV targeted therapies could be used to treat these malignancies. In this paper, we review a variety of different approaches currently under development that specifically target EBV-infected cells for destruction. EBV-based strategies for treating cancer include prevention of viral oncogene expression, inducing loss of the EBV episome, the purposeful induction of the lytic form of EBV infection, and enhancing the host immune response to virally encoded antigens. PMID- 12910251 TI - Cellular transformation by the HTLV-I Tax protein, a jack-of-all-trades. AB - The human T-cell leukemia virus type I (HTLV-I) is an oncogenic retrovirus that is responsible for adult T-cell leukemia and a neurological disease, HTLV-I associated myelopathy/tropical spastic paraparesis. HTLV-I encodes an oncogenic protein, Tax, which affects a variety of cellular functions prompting it to be referred to as a jack-of-all trades. The ability of Tax to both transcriptionally regulate cellular gene expression and to functionally inactivate proteins involved in cell-cycle progression and DNA repair provide the basis for Tax mediated transformation and leukemogenesis. This review will concentrate on the effects of Tax on the dysregulation of the G(1)/S and G(2)/M checkpoints as well as the suppression of DNA damage repair leading to cellular transformation. PMID- 12910252 TI - The role of Kaposi's sarcoma-associated herpesvirus/human herpesvirus-8 regulator of transcription activation (RTA) in control of gene expression. AB - The mechanisms that control the replication state, latency versus lytic, of human herpesviruses have been under intense investigations. Here we summarize some of the recent findings that help define such mechanisms for Kaposi's sarcoma associated herpesvirus/human herpesvirus type 8 (KSHV/HHV-8). For HHV-8, the viral regulator of transcription activation (RTA) is a key mediator of the switch from latency to lytic gene expression in infected cells. RTA is necessary and sufficient to drive HHV-8 lytic replication and the production of viral progeny. The RTA is an immediate-early gene product, it is the initial activator of expression of a multitude of viral and cellular genes that have been implicated in the replication of HHV-8 and pathogenesis of KS. Interactions of RTA with a number of viral promoters, and with a number of transcription factors or transcriptional co-activators are highlighted. Modulation of transactivation, through alternate RTA-protein, or RTA-promoter interactions, is hypothesized to participate in the selective tissue tropism and differential pathogenesis observed in KS. PMID- 12910254 TI - New developments about the association of SV40 with human mesothelioma. AB - Simian virus 40 (SV40) has been detected in human tumors in over 40 different laboratories. Many of these reports linked SV40 to human mesotheliomas. The Vaccine Safety Committee of the Institute of Medicine (IOM), National Academy of Sciences, USA, recently reviewed the evidence associating polio vaccines and/or SV40 with human tumors. The IOM conclusions about polio vaccines and human cancer were: (1) 'the evidence is inadequate to accept or reject a causal relation between SV40-containing polio vaccines and cancer' because the 'epidemiological studies are sufficiently flawed'; (2) 'the biological evidence is of moderate strength that SV40 exposure from the polio vaccines is related to SV40 infection in humans'. The epidemiological studies were considered flawed because it was not possible to distinguish reliably among exposed and nonexposed cohorts. Concerning SV40, the IOM concluded that (1) 'the evidence is strong that SV40 is a transforming virus; (2) the evidence is of moderate strength that SV40 exposure could lead to cancer in humans under natural conditions' (IOM, 2002). Similar conclusions were reached at an International consensus meeting on SV40 and human tumors held at the University of Chicago in 2001. G Klein and C Croce, who chaired the final panel that reviewed all the published evidence linking SV40 to human tumors, stated that 'the presence of SV40 in human tumors has been convincingly demonstrated' (Klein et al., 2002). In addition, a workshop organized by the Biological Carcinogenesis Branch of the National Cancer Institute, Bethesda, MD, chaired by J Pagano, has reached similar conclusions (Wong et al., 2002). Therefore, three independent scientific panels have all agreed that there is compelling evidence that SV40 is present in some human cancers and that SV40 could contribute to the pathogenesis of some of them. It should be noted that the presence of SV40 in mesothelioma and other human tumor types has been challenged by a research team that has consistently reported negative findings (Strickler et al., 2001). However, a member of this research team has recently acknowledged - in sworn testimony -sensitivity problems and possible irregularities that raise concerns about these negative reports (MacLachlan, 2002). These revelations, together with the conclusions of the three independent panels mentioned above, appear to bring to an end the apparent controversy about the presence of SV40 in human mesotheliomas and brain tumors. PMID- 12910253 TI - SV40 in human brain cancers and non-Hodgkin's lymphoma. AB - Simian virus 40 (SV40) is a potent DNA tumor virus that is known to induce primary brain cancers and lymphomas in laboratory animals. SV40 oncogenesis is mediated by the viral large tumor antigen (T-ag), which inactivates the tumor suppressor proteins p53 and pRb family members. During the last decade, independent studies using different molecular biology techniques have shown the presence of SV40 DNA, T-ag, or other viral markers in primary human brain cancers, and a systematic assessment of the data indicates that the virus is significantly associated with this group of human tumors. In addition, recent large independent studies showed that SV40 T-ag DNA is significantly associated with human non-Hodgkin's lymphoma (NHL). Although the prevalence of SV40 infections in humans is not known, numerous observations suggest that SV40 is a pathogen in the human population today. This review examines the molecular biology, pathology, and clinical data implicating SV40 in the pathogenesis of primary human brain cancers and NHL and discusses future research directions needed to define a possible etiologic role for SV40 in these malignancies. PMID- 12910255 TI - Human neurotropic polyomavirus, JCV, and its role in carcinogenesis. AB - A number of recent studies have reported the detection of the ubiquitous human polyomavirus, JC virus (JCV), in samples derived from several types of neural as well as non-neural human tumors. The human neurotropic JCV was first identified as the etiologic agent of the fatal demyelinating disease, progressive multifocal leukoencephalopathy, which usually occurs in individuals with defects in cell mediated immunity, including AIDS. However, upon mounting evidence of the oncogenic potential of the viral regulatory protein, T-antigen, and JCV's oncogenecity in a broad range of animal models, studies were initiated to determine its potential involvement in human carcinogenesis. Initially, the most frequently observed tumors in rodent models, including medulloblastoma, astrocytoma, glioblastoma, and other neural-origin tumors were analysed. These studies were followed by analysis of non-neural tumors such as colorectal carcinomas. In a subset of each tumor type examined, JC viral genomic DNA sequences could be detected by PCR and confirmed by Southern blot hybridization or direct sequencing. In a smaller subset of the tumors, the expression of T antigen was observed by immunohistochemical analysis. Owing to the established functions of T-antigen including its ability to interact with tumor suppressor proteins such as Rb and p53, and its ability to influence chromosomal stability, potential mechanisms of JCV T-antigen-mediated cellular dysregulation are discussed. Further, as increasing evidence suggests that T-antigen is not required for maintenance of a transformed phenotype, a hit-and-run model for T antigen-induced transformation is proposed. PMID- 12910256 TI - Oncogenic transformation by BK virus and association with human tumors. AB - BK virus (BKV), a human polyomavirus closely related to JC virus and Simian Virus 40, is ubiquitous in human populations worldwide. After primary infection, BKV establishes a lifelong latent infection in many organs. BKV transforms rodent cells to the neoplastic phenotype and is highly oncogenic in rodents. This review considers the oncogenic potential of BKV in humans and its possible involvement in human tumors. BKV sequences and T antigen (Tag) are detected in several types of human neoplasms, although the viral load is generally low, with less than one copy of the viral genome per cell. The possible causative role of BKV in human oncogenesis rests on the ability of BKV Tag to inactivate the functions of tumor suppressor proteins p53 and pRB family as well as on its ability to induce chromosomal aberrations in human cells. A 'hit and run' mechanism and secretion of paracrine growth factors by BKV Tag-positive cells, recruiting into proliferation neighboring and distant cells, are discussed as possible BKV pathogenic elements in human oncogenesis. PMID- 12910257 TI - Human papillomaviruses: targeting differentiating epithelial cells for malignant transformation. AB - Human papillomavirus (HPV) infections play a crucial role in the pathogenesis of cervical neoplasia. Insights into the mechanisms by which HPV infection can, in a small numbers of cases, result in malignancy, comes from the observation that three proteins encoded by high-risk genital HPVs, E6, E7 and to a lesser extent E5, target factors that control the cell cycle and proliferation. These interactions result in abrogation of cell cycle control, chromosomal alterations, telomerase activation, and eventual cell immortalization. In this review, we discuss the functions of E6, E7, and E5 proteins that are most relevant to the malignant progression of HPV-transformed cells. PMID- 12910258 TI - Cell cycle regulation and neural differentiation. AB - The general mechanisms that control the cell cycle in mammalian cells have been studied in depth and several proteins that are involved in the tight regulation of cell cycle progression have been identified. However, the analysis of which molecules participate in cell cycle exit of specific cell lineages is not exhaustive yet. Moreover, the strict relation between cell cycle exit and induction of differentiation has not been fully understood and seems to depend on the cell type. Several in vivo and in vitro studies have been performed in the last few years to address these issues in cells of the nervous system. In this review, we focus our attention on cyclin-cyclin-dependent kinase complexes, cyclin kinase inhibitors, genes of the retinoblastoma family, p53 and N-Myc, and we aim to summarize the latest evidence indicating their involvement in the control of the cell cycle and induction of differentiation in different cell types of the peripheral and central nervous systems. Studies on nervous system tumors and a possible contributory role in tumorigenesis of polyomavirus T antigen are reported to point out the critical contribution of some cell cycle regulators to normal neural and glial development. PMID- 12910259 TI - An alcohol dehydrogenase ribozyme. AB - We report an RNA molecule that exhibits activity analogous to that of alcohol dehydrogenase (ADH). Directed in vitro evolution was used to enrich nicotinamide adenine dinucleotide (NAD+)-dependent redox-active RNAs from a combinatorial pool. The most active ribozyme in the population forms a compact pseudoknotted structure and oxidizes an alcohol seven orders of magnitude faster than the estimated spontaneous rate. Moreover, this ADH RNA was coupled with a redox relay between NADH and flavin adenine dinucleotide to give a NAD+-regeneration system. Our demonstration of the redox ability of RNA adds support to an RNA-based metabolic system in ancient life. PMID- 12910260 TI - An mRNA structure that controls gene expression by binding S-adenosylmethionine. AB - Riboswitches are metabolite-binding RNA structures that serve as genetic control elements for certain messenger RNAs. These RNA switches have been identified in all three kingdoms of life and are typically responsible for the control of genes whose protein products are involved in the biosynthesis, transport or utilization of the target metabolite. Herein, we report that a highly conserved RNA domain found in bacteria serves as a riboswitch that responds to the coenzyme S adenosylmethionine (SAM) with remarkably high affinity and specificity. SAM riboswitches undergo structural reorganization upon introduction of SAM, and these allosteric changes regulate the expression of 26 genes in Bacillus subtilis. This and related findings indicate that direct interaction between small metabolites and allosteric mRNAs is an important and widespread form of genetic regulation in bacteria. PMID- 12910261 TI - Insights into the respiratory electron transfer pathway from the structure of nitrate reductase A. AB - The facultative anaerobe Escherichia coli is able to assemble specific respiratory chains by synthesis of appropriate dehydrogenases and reductases in response to the availability of specific substrates. Under anaerobic conditions in the presence of nitrate, E. coli synthesizes the cytoplasmic membrane-bound quinol-nitrate oxidoreductase (nitrate reductase A; NarGHI), which reduces nitrate to nitrite and forms part of a redox loop generating a proton-motive force. We present here the crystal structure of NarGHI at a resolution of 1.9 A. The NarGHI structure identifies the number, coordination scheme and environment of the redox-active prosthetic groups, a unique coordination of the molybdenum atom, the first structural evidence for the role of an open bicyclic form of the molybdo-bis(molybdopterin guanine dinucleotide) (Mo-bisMGD) cofactor in the catalytic mechanism and a novel fold of the membrane anchor subunit. Our findings provide fundamental molecular details for understanding the mechanism of proton motive force generation by a redox loop. PMID- 12910262 TI - Mesenchymal stem cells modified with Akt prevent remodeling and restore performance of infarcted hearts. AB - Transplantation of adult bone marrow-derived mesenchymal stem cells has been proposed as a strategy for cardiac repair following myocardial damage. However, poor cell viability associated with transplantation has limited the reparative capacity of these cells in vivo. In this study, we genetically engineered rat mesenchymal stem cells using ex vivo retroviral transduction to overexpress the prosurvival gene Akt1 (encoding the Akt protein). Transplantation of 5 x 10(6) cells overexpressing Akt into the ischemic rat myocardium inhibited the process of cardiac remodeling by reducing intramyocardial inflammation, collagen deposition and cardiac myocyte hypertrophy, regenerated 80-90% of lost myocardial volume, and completely normalized systolic and diastolic cardiac function. These observed effects were dose (cell number) dependent. Mesenchymal stem cells transduced with Akt1 restored fourfold greater myocardial volume than equal numbers of cells transduced with the reporter gene lacZ. Thus, mesenchymal stem cells genetically enhanced with Akt1 can repair infarcted myocardium, prevent remodeling and nearly normalize cardiac performance. PMID- 12910263 TI - Salusins: newly identified bioactive peptides with hemodynamic and mitogenic activities. AB - The discovery of endogenous bioactive peptides has typically required a lengthy identification process. Computer-assisted analysis of cDNA and genomic DNA sequence information can markedly shorten the process. A bioinformatic analysis of full-length, enriched human cDNA libraries searching for previously unidentified bioactive peptides resulted in the identification and characterization of two related peptides of 28 and 20 amino acids, which we designated salusin-alpha and salusin-beta. Salusins are translated from an alternatively spliced mRNA of TOR2A, a gene encoding a protein of the torsion dystonia family. Intravenous administration of salusin-alpha or salusin-beta to rats causes rapid, profound hypotension and bradycardia. Salusins increase intracellular Ca2+, upregulate a variety of genes and induce cell mitogenesis. Salusin-beta stimulates the release of arginine-vasopressin from rat pituitary. Expression of TOR2A mRNA and its splicing into preprosalusin are ubiquitous, and immunoreactive salusin-alpha and salusin-beta are detected in many human tissues, plasma and urine, suggesting that salusins are endocrine and/or paracrine factors. PMID- 12910264 TI - Bone marrow as a priming site for T-cell responses to blood-borne antigen. AB - Although bone marrow is known as a primary lymphoid organ, its potential to serve as a secondary immune organ has hardly been explored. Here we demonstrate that naive, antigen-specific T cells home to bone marrow, where they can be primed. Antigen presentation to T cells in bone marrow is mediated via resident CD11c+ dendritic cells. They are highly efficient in taking up exogenous blood-borne antigen and processing it via major histocompatibility complex class I and class II pathways. T-cell activation correlates with dendritic cell-T cell clustering in bone marrow stroma. Primary CD4+ and CD8+ T-cell responses generated in bone marrow occur in the absence of secondary lymphoid organs. The responses are not tolerogenic and result in generation of cytotoxic T cells, protective anti-tumor immunity and immunological memory. These findings highlight the uniqueness of bone marrow as an organ important for hemato- and lymphopoiesis and for systemic T cell-mediated immunity. PMID- 12910265 TI - CIITA-regulated plexin-A1 affects T-cell-dendritic cell interactions. AB - The major histocompatibility complex (MHC) class II transactivator (CIITA) is the 'master coactivator' of MHC class II genes. To identify new targets of CIITA, we analyzed cDNA microarrays of dendritic cells (DCs) from CIITA-deficient, MHC class II-deficient and control mice. We found the semaphorin receptor plexin-A1 was expressed in DCs, but not in other immune cells, and was strongly induced by CIITA. RNA interference by short hairpin RNA specific for plexin-A1, but not a single-nucleotide mutant, greatly reduced plexin-A1 expression and T cell stimulation by protein- or peptide-antigen-pulsed DCs.Plexin-A1 is not required for peptide binding to MHC. These data indicate involvement of plexin-A1 in T cell-DC interactions but not antigen processing or binding. PMID- 12910266 TI - Genetic tagging shows increased frequency and longevity of antigen-presenting, skin-derived dendritic cells in vivo. AB - Dendritic cells (DCs) are key regulators of immune responses that activate naive antigen-specific T lymphocytes. In draining lymph nodes, antigen-bearing DCs are reported to be rare and short-lived. How such small numbers of short-lived DCs can activate rare antigen-specific T cells is unclear. Here we show that after immunization of mouse skins by gene gun, the number of antigen-bearing DCs that migrate to draining lymph node is 100-fold higher than previously estimated and that they persist for approximately 2 weeks. The substantial frequency and longevity of DCs in situ ensures ample antigen presentation and stimulation for the rare antigen-specific T cells in draining lymph nodes. PMID- 12910267 TI - Defective development and function of Bcl10-deficient follicular, marginal zone and B1 B cells. AB - Bcl10 is an intracellular protein essential for nuclear factor (NF)-kappaB activation after lymphocyte antigen receptor stimulation. Using knockout mice, we show that absence of Bcl10 impeded conversion from transitional type 2 to mature follicular B cells and caused substantial decreases in marginal zone and B1 B cells. Bcl10-deficient B cells showed no excessive apoptosis. However, both Bcl10 deficient follicular and marginal zone B cells failed to proliferate normally, although Bcl10-deficient marginal zone B cells uniquely failed to activate NF kappaB efficiently after stimulation with lipopolysaccharide. Bcl10-deficient marginal zone B cells did not capture antigens, and Bcl10-deficient (Bcl10-/-) mice failed to initiate humoral responses, leading to an inability to clear blood borne bacteria. Thus, Bcl10 is essential for the development of all mature B cell subsets. PMID- 12910268 TI - AID mutant analyses indicate requirement for class-switch-specific cofactors. AB - Activation-induced cytidine deaminase (AID) is the essential and sole B cell specific factor required for class-switch recombination (CSR) and somatic hypermutation (SHM). However, it is not known how AID differentially regulates these two independent events. Involvement of several cofactors interacting with AID has been indicated by scattered distribution of loss-of-function point mutations and evolutionary conservation of the entire 198-amino-acid protein. Here, we report that human AID mutant proteins with insertions, replacements or truncations in the C-terminal region retained strong SHM activity but almost completely lost CSR activity. These results indicate that AID requires interaction with a cofactor(s) specific to CSR. PMID- 12910269 TI - Cidea-deficient mice have lean phenotype and are resistant to obesity. AB - The thermogenic activity of brown adipose tissue (BAT), important for adaptive thermogenesis and energy expenditure, is mediated by the mitochondrial uncoupling protein1 (Ucp1) that uncouples ATP generation and dissipates the energy as heat. We show here that Cidea, a protein of unknown function sharing sequence similarity with the N-terminal region of DNA fragmentation factors Dffb and Dffa, is expressed at high levels in BAT. Cidea-null mice had higher metabolic rate, lipolysis in BAT and core body temperature when subjected to cold treatment. Notably, Cidea-null mice are lean and resistant to diet-induced obesity and diabetes. Furthermore, we provide evidence that the role of Cidea in regulating thermogenesis, lipolysis and obesity may be mediated in part through its direct suppression of Ucp1 activity. Our data thus indicate a role for Cidea in regulating energy balance and adiposity. PMID- 12910270 TI - Association of cadherin 23 with polygenic inheritance and genetic modification of sensorineural hearing loss. AB - Age-related hearing loss (AHL) in common inbred mouse strains is a genetically complex quantitative trait. We found a synonymous single-nucleotide polymorphism in exon 7 of Cdh23 that shows significant association with AHL and the deafness modifier mdfw (modifer of deafwaddler). The hypomorphic Cdh23(753A) allele causes in-frame skipping of exon 7. Altered adhesion or reduced stability of CDH23 may confer susceptibility to AHL. Homozygosity at Cdh23(753A) or in combination with heterogeneous secondary factors is a primary determinant of AHL in mice. PMID- 12910272 TI - Chronotropic and dromotropic components of cardiac reflexes in rabbits. AB - The relationships between dromotropic and chronotropic components of five reflexes were studied in rabbits: intravenous and intraarterial blood injections, occlusion of the carotid arteries, Aschner maneuver, and stimulation of depressors. All these stimuli reduced heart rate (except carotid artery occlusion, which induced approximately equal number of tachi- and bradycardic responses). The former three stimuli also reduced atrioventricular (AV) conduction velocity, the changes in these two parameters were proportional. Changes in heart rate induced by Aschner maneuver were more pronounced than changes in AV conduction. Stimulation of depressor induced co-directed shifts in these parameters during the first seconds, but then AV conduction increased, while heart rate remained decreased; bradycardia and AV acceleration persisted for long time after termination of stimulation. Our findings attest to independent regulation of heart rate and AV conduction velocity and to the absence of a strict relationship between these two parameters. PMID- 12910273 TI - Involvement of rostral ventromedullar neuronal structures in nitric oxide modulation of central chemosensitive drive. AB - Surface perfusion of the rostral ventromedullar cerebral subdivisions with artificial cerebrospinal fluid containing exogenous NO donor sodium nitroprusside (0.1 mM) increased the discharge rate of the phrenic nerve and potentiated the response of the respiratory center to hypercapnia in narcotized mature rats. The latter reaction was prevented by blockage of NO-synthase in rostral ventromedullar neural structures with N(omega)-nitro-L-arginine methyl ester (L NAME, 0.3 mM). It was hypothesized that rostral ventromedullar neural structures are involved in modulatory action of NO on central chemosensitive drive. PMID- 12910271 TI - Comparative analysis of the genome sequences of Bordetella pertussis, Bordetella parapertussis and Bordetella bronchiseptica. AB - Bordetella pertussis, Bordetella parapertussis and Bordetella bronchiseptica are closely related Gram-negative beta-proteobacteria that colonize the respiratory tracts of mammals. B. pertussis is a strict human pathogen of recent evolutionary origin and is the primary etiologic agent of whooping cough. B. parapertussis can also cause whooping cough, and B. bronchiseptica causes chronic respiratory infections in a wide range of animals. We sequenced the genomes of B. bronchiseptica RB50 (5,338,400 bp; 5,007 predicted genes), B. parapertussis 12822 (4,773,551 bp; 4,404 genes) and B. pertussis Tohama I (4,086,186 bp; 3,816 genes). Our analysis indicates that B. parapertussis and B. pertussis are independent derivatives of B. bronchiseptica-like ancestors. During the evolution of these two host-restricted species there was large-scale gene loss and inactivation; host adaptation seems to be a consequence of loss, not gain, of function, and differences in virulence may be related to loss of regulatory or control functions. PMID- 12910274 TI - Respiratory activity of bulbospinal preparations from newborn rats subjected to periodic hypercapnia in the prenatal period. AB - Periodic exposure of pregnant rats to hypercapnia (10% CO(2)) delayed the development of mechanisms underlying respiratory rhythmogenesis in newborn animals during the early postnatal period. In vitro studies showed that rhythmic activity of the respiratory center in newborn rats (days 0-3) was 2-3-fold lower than that in intact preparations. Age-related changes of respiratory activity were absent, while the reactions of the respiratory center to CO(2) were suppressed compared to the control. PMID- 12910275 TI - Participation of endogenous opioids in pathogenesis of early neuroendocrine manifestations of prenatal stress syndrome. AB - We studied sex dimorphism in the content of norepinephrine and activity of enzymes involved in testosterone metabolism in the preoptic hypothalamic area of 10-day-old rats. Prenatal stress eliminated sex-related differences in these indices. These disturbances were absent in rats subjected to prenatal stress under conditions of opioid receptor blockade with naltrexone. These data attests to the important role of opioids in the pathogenesis of prenatal stress syndrome. PMID- 12910276 TI - Ketamine-induced rotational asymmetry in evaluation of motor disturbances in rats with middle cerebral artery occlusion. AB - Intraperitoneal injection of ketamine in subanesthetic doses to Wistar rats with unilateral occlusion of the middle cerebral artery caused ipsilateral rotation (2 10 rpm), which was recorded in an automatic rotameter. The optimal dose of ketamine was 50 mg/kg. The animals were examined in an automatic rotameter for 40 min. Motor asymmetry persisted for no less than 2 months after surgery. According to the neurological test (Menzies scale) motor asymmetry in animals with focal brain ischemia persisted for no more than 30 days. The degree of ketamine-induced motor asymmetry in intact rats was 0.10 0.03 rpm. PMID- 12910277 TI - DSLET and ACTH(4-10) increase mitotic activity of hepatocytes and suppress antibody production. AB - The mitotic index of hepatocytes remained unchanged after 10 intraperitoneal injections of DSLET and ACTH(4-10) in doses of 0.5, 1.5, and 5 microg/kg, but increased after injection of these substances in doses of 50 and 150 microg/kg. DSLET in doses of 5, 50, and 150 microg/kg decreased the number of antibody producing cells in the spleen. ACTH(4-10) possessed immunosuppressive activity not only in these doses, but also in a dose of 1.5 microg/kg. As differentiated from mitotic activity of hepatocytes, the degree of immunosuppression increased with increasing the dose of test peptides. PMID- 12910278 TI - Laser modification of the blood in vitro and in vivo in patients with Parkinson's disease. AB - The effect of He-Ne laser radiation on activity of MAO B, Cu/Zn-SOD, Mn-SOD, and catalase in blood cells from patients with Parkinson's disease was studied in vivo and in vitro. The effects of intravenous in vivo irradiation (intravenous laser therapy) were more pronounced than those observed in similar in vitro experiments. It is concluded that generalized effect of laser therapy involves interaction between blood cells. PMID- 12910279 TI - Modification of transport function of plasma albumin during atherosclerosis and diabetes mellitus. AB - Parameters characterizing the processes of association, transport, and dissociation of fatty acid molecules on the corresponding binding sites of plasma albumin in patients with atherosclerosis and diabetes mellitus were studied by electron paramagnetic resonance method. In these patients transport function of albumin differed from normal. It should be emphasized that these differences were specific for atherosclerosis and diabetes mellitus, which is of considerable diagnostic value. PMID- 12910280 TI - Dynamics of interrelationships between the content of lipoprotein particles, fibrinogen, and leukocyte count in the plasma from patients with coronary heart disease treated with Kwai. AB - The hypolipidemic effect of Kwai, a preparation based on garlic powder (Allium sativum), depended on the initial content of cholesterol and/or triglycerides. This effect was most pronounced in patients with coronary heart disease with initial cholesterol >7.0 mmol/liter and triglyceride >1.92 mmol/liter. After treatment with Kwai the correlation between triglyceride content and leukocyte count remained unchanged, the correlation between cholesterol content and leukocyte count disappeared, fibrinogen concentration decreased by 11%, and no correlation was found between fibrinogen content and leukocyte count. PMID- 12910281 TI - Pharmacogenetic study of anxiolytic effects of new cholecystokinin receptor antagonists in animals with different levels of emotionality. AB - Effects of two new peptide antagonists of central cholecystokinin receptors, GB 101 (0.05-0.40 mg/kg) and GB-115 (0.006-0.100 mg/kg), on the behavior of inbred animals differing by the reactions to emotional stress were studied in standard tests for evaluation of tranquilizers. The test drugs dose-dependently increased the total locomotor activity in the open field test in Balb/c mice and had no effect on the behavior of C57Bl/6 mice. GB-101 (0.4 mg/kg) and GB-115 (0.025 and 0.05 mg/kg) produced an anxiolytic effect on MR rats (but not on MNRA rats) in the elevated plus-maze and conflict tests. These data confirm anxioselective effects of the test compounds. PMID- 12910282 TI - Qualitative assessment of selective blockade of M(4)-cholinoreceptors in the whole organism. AB - Quantitative assessment of selective blockade of M4-subtype muscarinic receptors was performed by the number of pilocarpine-induced movements of lower jaw in rats. Three antagonists (atropine, cyclodol, and glipin) were used in the experiments. Glipin produced the most potent blockade of M4 receptors in the whole organism compared to other test antagonist. PMID- 12910283 TI - Characteristics of collagenolytic enzymes secreted by deuteromycete fungi Aspergillus flavus. AB - Collagen induced the synthesis of collagenolytic enzymes in the culture of Aspergillus flavus. Enzyme activity of the culture increased during storage and passage on a medium with this inductor. We developed a method for isolation and purification of collagenolytic enzymes and obtained two electrophoretically homogenous enzyme preparations belonging to neutral thermolabile collagenolytic metalloproteinases. PMID- 12910284 TI - Stimulation of mouse resistance to bacterial infection with muramyl dipeptide glycoside. AB - We studied the capacity of 9 new muramyl dipeptide glycosides to stimulate mouse resistance to experimental sepsis induced by intraperitoneal injection of salmonella typhimurium culture. Preventive intraperitoneal injections of muramyl dipeptide beta-glycosides better improved survival of infected animals compared to the original (unmodified) muramyl dipeptide and muramyl dipeptide alpha glycosides. The most effective drug muramyl dipeptide beta-heptylglycoside injected during sepsis development also reduced animal mortality, decreased bacterial contamination of the viscera, and increased phagocytic activity of peritoneal macrophages in infected animals. PMID- 12910285 TI - Adaptation variability of Yersinia pseudotuberculosis during long-term persistence in soil. AB - Adaptation changes in the morphological and biological characteristics of pathogenic Yersinia pseudotuberculosis inhabiting a model soil ecosystem for a long time were studied. Changes in cultural, biochemical, and morphological characteristics of Yersinia and a decrease or loss of virulence were observed under these conditions. The detected changes were phenotypical, i. e. several passages in common nutrient media restored the original characteristics of the strain. PMID- 12910286 TI - IgE and IgG antibodies to Malassezia spp. yeast extract in patients with atopic dermatitis. AB - The presence of immunoglobulins to Malassezia spp. surface proteins in the sera from patients with atopic dermatitis and healthy subjects was studied. It was found that 28% of 25 examined patients with atopic dermatitis had IgE antibodies to Malassezia spp. surface protein preparation. All patients and 5 healthy subjects had IgG antibodies to this preparation. The presence and concentration of specific IgE antibodies in patients with atopic dermatitis correlated with reverse titers of IgG antibodies to this preparation (r=0.782). The medians of values reciprocal to IgG antibody titers in patients with atopic dermatitis with and without specific IgE antibodies to the preparation and in healthy subjects were 64, 1024, and 16, respectively. The preparation derived from Candida albicans (candidine) and previously derived preparation from Malassezia did not cross-react. According to immunoblotting data, the preparation contains allergens presented by proteins with molecular weights 15, 36, 52-56, and 78.4 kDa. PMID- 12910287 TI - Destructive effect of DNA topoisomerase II inhibitor vepesid on mouse spermatogenesis. AB - Inhibition of DNA topoisomerase II with vepesid induced structural and functional reorganization of chromatin in meiotically dividing spermatocytes I, which later led to the block of their differentiation and long-lasting disorders in spermatogenesis. Vepesid induced decondensation of spermatocyte I chromatin, block of desynapsis, and elongation of lateral elements of spermatocyte autosome synaptonemal complexes during late pachytene and diplotene of meiosis. This confirms the involvement of type II DNA topoisomerase in chromatin condensation and homologous chromosome desynapsis at the stage of diplotene and the role of this enzyme in structural organization of the synaptonemal complex. Vepesid induced the formation of dichotomy and breaks of the pericentromer regions of subelements of lateral elements of the autosomal synaptonemal complexes; the number of cells with associations of axial elements of sex chromosomes with autosomal synaptonemal complexes increased, univalents of autosomes and sex chromosomes appeared. Mesna, a modifier of toxic effects of antitumor drugs, had no toxic effect on spermatogenic cells. Mesna reduced the lethal effect of vepesid during combined treatment, but did not ensure long-term protection of spermatogenesis. PMID- 12910288 TI - Transcription regulatory cis-element (GCC)8 in the 5'-untranslated region of the gene for human very-low-density lipoprotein receptors. AB - The 5'-untranslated region of very-low-density lipoprotein (VLDL) receptor gene includes 2 groups of triplet repeats (GCC)n. Four repeats are localized near the promoter region in position 15.23 from the transcription initiation site. Eight repeats were detected in position 573.597. Sequence (GCC)8 in VLDL receptor gene forms specific complexes with nuclear proteins of HepG2 cells, the formation of these complexes depended on Zn(2+). Superexpression of the CGGBP-20 protein interacting with long sequences (GCC)n suppressed transcriptional activity of VLDL receptor gene. Removal of fragment (397.616) containing cis-element (GCC)8 from the 5'-untranslated region of VLDL receptor gene led to activation of the linked marker gene cat in Hutu80 cells, but did not abolish the repressor effect of CGGBP-20 protein. Our results suggest that (GCC)n-binding proteins differing from CGGBP-20 regulate activity of the VLDL receptor gene via cis-element (GCC)8. PMID- 12910289 TI - Effect of metabolic factors on apoptosis in thymocytes during tumor growth. AB - Intensive apoptotic death of thymocytes is a possible mechanism of thymus involution during tumor growth. We studied the role of hypercholesterolemia and lactate acidosis in the induction of increased sensitivity of thymocytes to apoptosis during growth of transplanted hepatoma 22a in mice. Spontaneous apoptosis in thymocytes during tumor growth in mice was studied in vitro by acridine orange/ethidium bromide staining and diphenylamine test. Plasma levels of lactate, total cholesterol, alpha-cholesterol, and triglycerides were measured. A positive correlation was found between intensification of apoptosis (diphenylamine test) and increased concentration of total plasma cholesterol on days 21 and 28 after inoculation of tumor cells. Plasma lactate content did not increase at this term. We hypothesize that hypercholesterolemia accompanying tumor growth acts as a factor increasing thymocyte sensitivity to apoptosis. PMID- 12910290 TI - Expression of Flt-1 and Flk-1 receptors for vascular endothelial growth factor on tumor cells as a new prognostic criterion for locally advanced breast cancer. AB - We studied expression of Flt-1 and Flk-1 receptors on tumor cells obtained from 83 patients with locally advanced breast cancer after neoadjuvant chemotherapy. The mean period of observations was 32.3 months. The median recurrence-free survival periods for Flt-1(+) and Flt-1(-) patients were 55 and 32 months, respectively (p=0.0064). The overall survival periods for Flt-1(-) and Flt-1(+) patients were 45 and 67.6 months, respectively (p=0.014). The mean recurrence free survival periods for Flk-1(+) and Flk-1(-) patients were 40.8 and 60.9 months, respectively (p=0.035). Expression of VEGF had no prognostic value. Our results show that overexpression of Flk-1 on breast cancer cells in patients receiving neoadjuvant chemotherapy is associated with a poor prognosis. By contrast, overexpression of Flt-1 improves survival. PMID- 12910291 TI - New approach to vital analysis of functional activity in ABC transporters (markers for multidrug resistance) in solid tumors by the method of flow cytofluorometry. AB - We developed and described a new approach to vital analysis of functional activity of multidrug resistance markers (ABC transporters) in intact biopsy specimens from human solid tumors by the method of flow cytofluorometry. The algorithm of the study underwent revision, and the cell suspension was obtained in the final stage. Intensification of intracellular doxorubicin accumulation (fluorescence) and increase in the number of fluorescent cells and total fluorescence of doxorubicin-accumulating cells produced by ABC transporter inhibitor sodium azide served as the criteria of expression of these transporters in tumor tissue. Informative value of changes in various parameters of doxorubicin fluorescence is discussed. The increase in the count of fluorescent cells in the suspension of tumors cells after treatment with the inhibitor indicates the presence of tumor cells absolutely resistant to this preparation. The proposed method is technically simple, suitable for structurally different tumors, requires small amounts of the biopsy material and, therefore, can be used for routine analysis. The results of our analysis and spectrofluorometric assay of ABC transporters agree very closely, which suggest that this method is adequate for the purpose. PMID- 12910292 TI - Immunohistological study of NM 23 and C-erbB-2 expression in primary tumor and metastases of colorectal adenocarcinoma. AB - Immunohistochemical study of expression of nm 23 and c-erbB-2 protein markers in primary tumor and metastases of colorectal adenocarcinoma showed that the incidence and expression of both protein markers in primary tumor tissue tended to increase after the appearance of liver metastases. Peculiarities of protein expression in tumor metastases in lymph nodes and liver and the possibility of using these markers as additional prognostic criteria in colorectal cancer are discussed. PMID- 12910293 TI - Retinoprotective effect of Epithalon in campbell rats of various ages. AB - We studied the retinoprotective effect of Epithalon administered to the offspring of Campbell rats during postnatal ontogeny and to mothers before mating and during pregnancy. After this treatment the morphological structure and functional activity of the retina were preserved for a longer period compared to control rats (by 2 times) and to the animals receiving the peptide only during postnatal ontogeny (by 30%). PMID- 12910294 TI - Effect of medical and biological factors on neurological manifestations of vertebral osteochondrosis in residents of the southern Altai Mountains. AB - We performed clinical and epidemiological study of 1508 residents living in the southern Altai Mountains and belonging to two subethnic groups (Telengite and Altai Kizhi). The incidence of neurological manifestations of vertebral osteochondrosis in people older than 17 years was 627.6 26.4%. The results show that genetic factors (hereditary polygenic predisposition) play a major role in the development of neurological manifestations of vertebral osteochondrosis. Premorbid state of the organism and diseases of various organs and systems promote the development of this neurological disorder. PMID- 12910295 TI - Pathomorphology of the vascular bed of postinfarction heart in various types of remodeling. AB - We evaluated pathomorphological changes in the vascular bed of postinfarction heart in various types of remodeling. Dilatational remodeling was characterized by dilation of coronary arteries, increase in their volume density, and regular arrangement. Signs of coronary blood flow reduction and microcirculatory disturbances in the left ventricle were revealed during hypertrophic remodeling. Aneurysmal remodeling was characterized by the presence of small-vascular collateral-anastomotic plexus and reduction of the microcirculatory bed. Endocardial remodeling was accompanied by hypervascularization of the myocardium, formation of new coronary vessels, and reduced capillarization in the left ventricle of postinfarction heart. PMID- 12910296 TI - Ultrastructural characteristics of hepatocytes during echinococcosis. AB - We performed an electron microscopy study and histological examination of hepatocytes during hepatic echinococcosis. The type of damage to liver cells depended on intrahepatic localization of hydatid cysts, which determined the development of postoperation complications. PMID- 12910297 TI - Chronic hepatitis C: modern notions of pathogenesis and morphogenesis. Concept of antiviral protection in hepatocytes. AB - Here we review modern notions about biological properties of hepatitis C virus, geographic distribution of its genotypes, peculiarities of immune reactions during chronic HCV infection, and role of viral replication in the infectious process. The concept of antiviral protection in liver parenchymal cells suggests suppression of biosynthetic reactions in hepatocytes resulting in inhibition of viral replication and focal degradation of the cytoplasm in infected cells followed by exocytosis and elimination of viral particles (cytosanation without cytodestruction). The recovery of structural characteristics in hepatocytes is associated with intracellular regeneration. Special accent is placed on reversibility of liver fibrosis in chronic HCV infection due to resorption of collagen fibers by hepatocytes. These data indicate that the therapy of liver fibrosis holds much promise. PMID- 12910299 TI - Mechanism of synergism between sympathetic and parasympathetic autonomic nervous systems in the regulation of motility of the stomach and sphincter of Oddi. AB - The mechanisms of stimulatory effect of the sympathetic trunk on gastric motor activity and vagal inhibitory effect on electromotor activity of the sphincter of Oddi were studied. Gastric contractions were augmented by preganglionic serotoninergic fibers related synaptically to serotoninergic neurons, while inhibition of electromotor activity of the sphincter of Oddi was elicited by activation of alpha- and beta-adrenoceptors. PMID- 12910298 TI - Dynamics of swallowing-induced cardiac chronotropic responses in healthy subjects. AB - Simultaneous recording of ECG and swallowing movements in healthy humans (n=23, age 20-57 years) showed that each swallow is accompanied by transient tachycardia with initial abrupt and pronounced heart rate increase. These rapid changes in heart rate (evaluation by maximum increment of heart rate over two successive heartbeats, Delta HR(2bt)) are typical of vagal chronotropic responses. The amplitude of tachycardia induced by a single swallow was significantly higher in the supine position (13.1 +/- 5.6 bpm) compared to the standing position (8.5 +/- 3.8 bpm; p<0.0001). Chronotropic responses to a series of three or more successive swallows consisted of two phases, the initial abrupt acceleration and subsequent slower growth of heart rate. In the standing position, the portion of the first rapid phase significantly decreased, while the portion of the slower phase increased compared to the supine position. The amplitude of tachycardia induced by a single swallow and parameter Delta HR(2bt) can serve as indices of the strength of parasympathetic modulation of the heart. By contrast, further slow increase in the heart rate determined by summation of responses to a series of successive swallows can result from not only inhibition of the parasympathetic influences, but also enhancement of sympathetic activity during swallowing. PMID- 12910300 TI - Whether independent regulation of myocardial contractility and diastolic relaxation rate is possible? AB - In acute experiments on cats neural inotropic and lusitropic reactions of the heart to enhancement of pre- and afterload were assessed by changes in contractility and relaxation indices, which were preliminary chosen for their maximum specificity and sensitivity. The control cardiac responses to increased pre- and afterload were measured after treatment with ganglionic blocker arfonad. The myogenic component of these responses assessed under the action of arfonad was highly pronounced, therefore the neural inotropic and lusitropic reactions were measured as the difference between load-induced changes of indices in experiments with and without arfonad. Increased preload produced similar negative inotropic and lusitropic effects, while increased afterload produced a more pronounced negative inotropic effect, which indicated independent regulation of contractility and diastolic relaxation of the heart. PMID- 12910301 TI - Effect of glyprolines on norepinephrine tone of isolated rat aortic rings. AB - Glyprolines, short proline-containing peptides (Pro-Gly-Pro, Pro-Gly, and Gly Pro) in concentrations of 10(-8)-10(-4) M decreased norepinephrine tone of the isolated rat aortic rings. The vasodilatory effect of Pro-Gly-Pro and Pro-Gly was associated with NO release from endotheliocytes, while Gly-Pro directly affected vascular smooth muscle cells. PMID- 12910302 TI - Neurosis of acquired helplessness and role of hypoxia in the formation of this disorder in rats. AB - Acquisition of instrumental defense response with pain reinforcement uncertainty (25% reinforcement) induced the development of acquired helplessness in 50% rats. Acquired helplessness is characterized by the absence of responses to conditioned (light) and unconditioned stimuli (pain), minor response of plasma corticosterone to learning, gas markers of circulatory cerebral hypoxia (Delta A/V pO2 carotid artery/jugular vein), low sensitivity to severe hypobaric conditions, and high resistance of Purkinje cells in the cerebellum. Piracetam improved learning and prevented the development of acquired helplessness. Local changes in cerebral blood flow and energy deficit in neurons responsible for emotional stress during acquired helplessness impair adaptive capacity, but reduce energy consumption and protect neuronal structures. PMID- 12910303 TI - Posthypoxic disturbances in glutamatergic signal transduction in rat brain neurons: correction effect of preconditioning. AB - Severe 3-h hypobaric hypoxia was followed by impairment of Ca(2+)-mediated glutamatergic signal transduction in the posthypoxic period (no less than 72 h). This impairment manifested in changes in the calcium response to glutamate application in slices of rat brain cortex. Moderate hypoxic preconditioning prevented these disturbances developed over the first day after sever hypoxia. PMID- 12910304 TI - LPO and antioxidant defense in the stomach of albino rats injected with angiotensin II and enalapril maleate. AB - The effects of components of angiotensin II system on LPO and antioxidant defense in the stomach of adult albino rats were studied using biochemical and chemiluminescent methods. Five intraperitoneal injections of angiotensin II in a dose of 100 micro/kg activated LPO and inhibited antioxidant processes in the studied tissues. Oral therapy with enalapril maleate (inhibitor of angiotensin converting enzyme) in a daily dose of 10 mg/kg for 2 weeks normalized stress activated LPO processes in gastric tissue. PMID- 12910305 TI - Changes in blood biochemistry in mice during development of experimental depression. AB - Biochemical parameters of the plasma were studied in mice at different stages of the development of depression-like states in males after social defeats in 10 and 20 intermale confrontations (T10 and T20 victims, respectively). Glucose and cholesterol levels were increased in T10 victims in comparison with intact animals. In T20 victims the increase in glucose level was paralleled by an increase in total protein. T20 victims differed from T10 victims by lower catalase activity. PMID- 12910306 TI - Tumor necrosis factor-alpha in low doses preactivates and activates macrophages by increasing their ability to produce reactive oxygen species and oxidize low density lipoproteins: protective effect of antioxidants. AB - Tumor necrosis factor-alpha in low doses activated rat peritoneal macrophages and intensified production of reactive oxygen species (zymosan-depended chemiluminescence). Single or 2-fold incubation with tumor necrosis factor-a activated and preactivated human blood macrophages and promoted oxidative modification of low-density lipoproteins (increased their mobility in agarose gel). Antioxidants (potassium phenosan, probucol, and desferal) suppressed oxidative modification of low-density lipoproteins induced by nonactivated, preactivated, and activated macrophages. Our results show that antioxidants hold much promise for the prevention and therapy of atherosclerosis. PMID- 12910307 TI - Antioxidant status of rats receiving lycopene in different doses. AB - Oral treatment with lycopene (per os) in doses of 10 or 50 mg/kg for 2 weeks led to accumulation of lycopene in the liver, liver microsomes, and blood plasma, increased total plasma antioxidant activity, inhibited LPO in the liver, and decreased solubilization of lysosomal enzymes. Lycopene had no effect on ex vivo resistance of liver microsomes to LPO and activities of antioxidant enzymes in the liver. PMID- 12910308 TI - Effect of sodium succinate on gas exchange in rats with barbiturate-induced coma. AB - Injection of sodium succinate in doses of 5 or 10 mmol/kg (but not 1 mmol/kg) intensified oxygen consumption in rats with sodium thiopental-induced coma. Injection of SDH inhibitor (sodium malonate) inhibited gas exchange and abolished the effect of sodium succinate. The effect of succinate on rat survival was positive, while that of malonate was negative, but manifested only as a trend. The critical role of succinate oxidation in preventing lethal complications of barbiturate-induced coma is proved. PMID- 12910309 TI - Metabolism of PGP peptide after administration via different routes. AB - Main pathways of degradation of PGP peptide possessing antiulcer and antithrombotic activities were studied after its intraperitoneal, intragastric, and intraintestinal administration. In experiments on rabbits we showed by HPLC that unmodified PGP is released into the blood after administration by all three routes and is detected in the plasma over 3-5 h. PG dipeptide is a more stable PGP metabolite presumably determining (together with tripeptide) its pharmacological properties. PMID- 12910310 TI - Sensitivity of immunocompetent cells of DBA/2 and C57Bl/6 mice to cyclophosphamide. AB - T cells were most sensitive to cyclophosphamide in DBA/2 mice, while in C57Bl/6 mice both T and B cells were sensitive. The formation of antibody-producing cells and the production of specific antibodies were delayed in DBA/2 mice immunized after pretreatment with antitumor drug. Accumulation of antibody-producing cells in the spleen was more active in immunized C57Bl/6 mice treated with cyclophosphamide compared to animals not treated with cyclophosphamide. PMID- 12910311 TI - Arrhythmogenic effects of endothelin-1 under conditions of NO-synthase blockade with L-NAME in NMRI mice. AB - Arrythmogenic effects of endothelin-1 were studied in NMRI mice under conditions of NO-synthase blockade with N omega-nitro-L-arginine methyl ester. Intravenous injection of endothelin-1 increased heart rate variability in awake mice. NO synthase blockade potentiated the arrythmogenic effects of endothelin-1. In narcotized animals the arrythmogenic effect of endothelin-1 was not observed and was considerably weakened under conditions of NO-synthase blockade. Arrhythmia was paralleled by atrioventricular block and lengthening of the ST segment. PMID- 12910312 TI - IL-5 expression in the sputum of patients with bronchial asthma. AB - Expression of IL-5 mRNA and the content of IL-5 in the sputum of patients with asthma of different severity were studied before and after treatment. The expression of IL-5 mRNA in mild asthma differed from that in severe and moderate asthma before and after treatment. The level of IL-5 before therapy was different in patients with mild and severe disease. In patients with severe asthma the level of IL-5 differed before and after treatment. PMID- 12910313 TI - Secretion of proinflammatory cytokines by villous chorion tissue in spontaneous abortion. AB - Secretion of proinflammatory cytokines IL-1 beta, TNF-alpha, IL-6, IFN-alpha, and IFN-gamma by the chorion tissue (8-12 weeks) was studied in normal gestation and spontaneous abortion. The production of IL-1 beta, TNF-alpha, and IFN-alpha virtually did not change in spontaneous abortion, while IFN-gamma was not secreted in all experimental groups. The production of IL-6 increased more than 2 fold in patients with spontaneous abortion during the first trimester. These data confirm the involvement of this cytokine in the reproductive processes and in the pathogenesis of miscarriage. PMID- 12910314 TI - Role of dietary starches in induction of immune cells apoptosis in Wistar rats. AB - We compared the effects of maize and potato starch on Wistar rats. The level of oligonucleosomal DNA fragmentation increased in thymic and splenic cells of animals receiving potato starch. The expression of Fas/CD95/Apo-1 apoptosis receptor in lymphocytes and their response to exogenous ceramide was also higher in rats receiving potato starch compared to the reference group. This led to intensification of spontaneous and induced apoptosis and decreased cell content in immune organs. The authors conclude that higher resistance of potato starch to digestive enzymes leads to modification of cell metabolism and induction of ceramide-dependent apoptosis in immune cells. PMID- 12910315 TI - Photodynamic inhibition of infection caused by drug-resistant variants of herpes simplex virus type I. AB - Membranotropic amphiphilic chromophore merocyanine 540 sensitized photodynamic inhibition of drug-resistant and sensitive variants of type I herpes simplex virus in cultured Vero cell. Optimal conditions of photodamage to virus particles and infected cells were determined (merocyanine 540 concentration 1 microM, illumination dose 32.5-65.0 kJ/m(2), exposure at early stages of infection). Infected cells actively bind the photosensitizer, which explains their selective photodamage. PMID- 12910316 TI - Nociceptive thresholds in adult rats of three strains after pain stimulation in the neonatal period. AB - Nociceptive thresholds in the tail-flick test decreased in adult 2.5-month-old KM, Wag/Rij, and Wistar rats receiving injections of placebo in the neonatal period (days 3-9 of life). PMID- 12910317 TI - Central non-NMDA receptors of a honey bee with hereditary kynurenine deficiency. AB - Pharmacological characteristics of non-NMDA receptors involved in associative learning were studied in wild honey bees (normal) and carriers of snow laranja mutation (kynurenine deficiency) by pharmacological analysis and behavioral criteria. The effects of systemic injections of non-NMDA receptor agonists (AMPA, kainic, quisqualic, and domoic acids), AMPA receptor antagonist (NS257-HCl), and AMPA receptor modulator (cyclothiaside) on retention of conditioned reflexes in short-term memory (1 min after the end of learning) were studied. The pharmacological characteristics of non-NMDA receptors were changed in snow laranja mutants with kynurenine deficiency. PMID- 12910318 TI - Differentiation of bone marrow stromal stem cells into cardiomyocyte-like cells in different mammalian species. AB - We describe the possibility of obtaining cardiomyocyte-like cell cultures from rat, guinea pig, and human bone marrow stromal stem cells. The content of troponin I-positive cells attains 35-45% of the total number of cells in the cultures and persists at this level for up to 4 months under differentiation conditions. Spontaneous contractions of cardiomyocyte-like cells were observed after the formation of cell monolayer under differentiation conditions. PMID- 12910319 TI - Xenotransplantation of stem/progenitor cells from human fetal brain to adult rats with spinal trauma. AB - In vitro grown neural stem cells from human fetal brain were transplanted to adult rats with spinal trauma. The spinal cord was examined morphologically using histological and immunohistochemical methods on days 5, 15, 30, and 110. Human neural stem/progenitor cells were viable, migrated, and differentiated into neurons and glia in the traumatized spinal cord in adult rats. PMID- 12910321 TI - Structure of human vertebral lamellar bone in age-associated involution and osteoporosis. AB - We compared the structure of lamellar bone in anterolateral zones of vertebral bodies in adult and senile subjects in health and osteoporosis. The appearance of structural signs of age-related osteopenia and enlargement of coplanar consolidations of the bone matrix crystals were noted. Osteopenia was more pronounced in osteoporosis, while the structural organization of the mineral component did not differ from that in age-matched controls. PMID- 12910320 TI - Effect of ascovertin on morphofunctional changes in the ovaries in monopolar coagulation. AB - A course of ascovertin therapy reduced manifestations of perifocal inflammation and atresia of growing follicles, corrected vascular disorders and changes in the tissue volume ratios outside the necrotic zone after monopolar coagulation of the ovaries in rats. The drug prevented the increase in blood viscosity, erythrocyte aggregation, and LPO activation in the early postoperation period. PMID- 12910322 TI - Lustra, Lustra-AF and Alustra. AB - Hydroquinone has been successfully used to treat hyperpigmentation disorders for many years. Recently, new formulations containing hydroquinone have become available, including Lustra and Lustra-AF (Medicis). These products also contain glycolic acid 2%, an active antioxidant system (ascorbyl palmitate and tocopherol acetate), and moisturizers. Lustra-AF also contains a broad-spectrum sunscreen. Alustra contains a stabilized, high-concentration of retinol. The above formulations inhibit melanogenesis, stimulate epidermal desquamation, inhibit free radical-mediated photodamage and restore the antioxidant reservoir. The addition of retinoids may facilitate epidermal penetration of hydroquinone and prevent its oxidation. Comparative studies have shown that these agents can be effective in reducing blotchiness, mottled hyperpigmentation, post-inflammatory hyperpigmentation, and surface roughness. In addition, these formulations have been generally well tolerated with patients rarely reporting mild-to-moderate adverse events such as dryness, redness, or peeling of the skin. PMID- 12910323 TI - Treatment of hand eczema. AB - Hand eczema is a common skin disease that tends to become chronic and may interfere with many types of work. Emollients have been shown to be useful in reducing eczema activity and in the primary prevention of hand eczema. Protection of the hands is very important for the prevention of hand eczema and is a fundamental aspect of the treatment of hand eczema. Although topical corticosteroids are the mainstay of treatment, few studies of their rational use, efficacy and side-effects have been conducted. A combination of tacrolimus and topical corticosteroids may reduce the risk of steroid-associated side-effects. Systemic treatment with immunosuppressants such as cyclosporine and methotrexate show promising results, and acitretin may suppress keratotic hand eczema. Botulinum toxin has been used with success in the treatment of dyshidrotic hand eczema. PUVA is effective as a systemic treatment. Bath-PUVA treatment, UVB and Grenz rays can also suppress hand eczema. PMID- 12910324 TI - Using banana to generate lactic acid through batch process fermentation. AB - We evaluated the usefulness of waste banana for generating lactic acid through batch fermentation, using Lactobacillus casei under three treatments. Two treatments consisted of substrates of diluted banana puree, one of which was enriched with salts and amino acids. The control treatment comprised a substrate suitable for L. casei growth. When fermentation was evaluated over time, significant differences (P<0.05) were found in the three treatments for each of five variables analyzed (generation and productivity of lactic acid, and consumption of glucose, fructose, and sucrose). Maximum productivity was (in g l( 1) h(-1)) 0.13 for the regular banana treatment, 1.49 for the enriched banana, and 1.48 for the control, with no significant differences found between the latter two treatments. Glucose consumption curves showed that L. casei made greater use of the substrate in the enriched banana and control treatments than in the regular banana treatment. For fructose intake, the enriched banana treatment showed significantly better (P<0.05) results than the regular one. Sucrose consumption was insignificant (P<0.05), probably because fermentation time was too short. Even when enriched, diluted banana puree is an ineffective substrate for L. casei, probably because it lacks nutrients. PMID- 12910325 TI - Acetone butanol ethanol (ABE) production from concentrated substrate: reduction in substrate inhibition by fed-batch technique and product inhibition by gas stripping. AB - Acetone butanol ethanol (ABE) was produced in an integrated fed-batch fermentation-gas stripping product-recovery system using Clostridium beijerinckii BA101, with H(2) and CO(2) as the carrier gases. This technique was applied in order to eliminate the substrate and product inhibition that normally restricts ABE production and sugar utilization to less than 20 g l(-1) and 60 g l(-1), respectively. In the integrated fed-batch fermentation and product recovery system, solvent productivities were improved to 400% of the control batch fermentation productivities. In a control batch reactor, the culture used 45.4 g glucose l(-1) and produced 17.6 g total solvents l(-1) (yield 0.39 g g(-1), productivity 0.29 g l(-1) h(-1)). Using the integrated fermentation-gas stripping product-recovery system with CO(2) and H(2) as carrier gases, we carried out fed batch fermentation experiments and measured various characteristics of the fermentation, including ABE production, selectivity, yield and productivity. The fed-batch reactor was operated for 201 h. At the end of the fermentation, an unusually high concentration of total acids (8.5 g l(-1)) was observed. A total of 500 g glucose was used to produce 232.8 g solvents (77.7 g acetone, 151.7 g butanol, 3.4 g ethanol) in 1 l culture broth. The average solvent yield and productivity were 0.47 g g(-1) and 1.16 g l(-1) h(-1), respectively. PMID- 12910326 TI - Production of nisin with continuous adsorption to Amberlite XAD-4 resin using Lactococcus lactis N8 and L. lactis LAC48. AB - The production of nisin, biomass and lactic acid in pH-controlled and uncontrolled batch fermentation and batch fermentation (pH 5.5) with continuous removal of nisin was examined in the parent strain Lactococcus lactis N8 and LAC48. Strain LAC48 in batch fermentor (pH not controlled) gave a maximum nisin concentration of 2.5 x 10(6) IU g dcw(-1). The nisin concentration remained high (2.0 x 10(6) IU g dcw(-1)) after the logarithmic growth phase (10-22 h), whereas nisin production of strain N8 decreased after the logarithmic growth phase. The maximum nisin production of strain LAC48 was not directly related to the biomass formation and not associated with growth. In order to study end product inhibition in nisin production, a system was built for adsorption of nisin during fermentation. The adsorbent Amberlite XAD-4 was found to have an effective binding capacity for nisin. Cells of LAC48 and N8 compensated for the removal of nisin, indicating that nisin production also occurs in the stationary phase. PMID- 12910327 TI - Response to different environmental stress conditions of industrial and laboratory Saccharomyces cerevisiae strains. AB - Two sets of Saccharomyces cerevisiae strains were compared for their physiological responses to different stress conditions. One group is composed of three strains adapted to controlled laboratory conditions (CEN.PK, LR88 and RS58), whereas the other consisted of five industrial strains (IND1101, SuperStart, LO24, LO41 and Azteca). Most industrial strains showed higher tolerance to heat shock and to an oxidative environment than laboratory strains. Excluding CEN.PK, a similar behavior was observed regarding ethanol production in high sugar concentrations (180 g/l glucose). Addition of acetate (10 g/l) or furfural (2 g/l), in concentrations similar to those found in sugar cane bagasse hydrolysates, decreased cell mass formation and growth rate in almost all strains. CEN.PK and SuperStart showed the highest sensitivity when grown in furfural-containing medium. Acetic acid treatment severely affected cell mass formation and reduced growth rate in all strains; CEN.PK and LO24 were the most resistant. The specific ethanol production rate was not affected by furfural addition. However, specific ethanol production rates decreased in response to acetic acid in four industrial strains, and increased in all laboratory strains and in LO24. No significant correlation was found between the stress tolerance of the strains tested and the transcript accumulation of genes selected by their involvement in the response to each of the stressful environments applied. PMID- 12910328 TI - Bioaugmentation of the phyllosphere for the removal of toluene from indoor air. AB - The removal of airborne toluene by means of the phyllosphere of Azalea indica augmented with a toluene-degrading enrichment culture of Pseudomonas putida TVA8 was studied. The 95% disappearance time [DT95%; the time in which an initial toluene concentration of 90 ppmv (339 mg.m(3)) was removed in a batch experiment] was 75 h for Azalea plants. Under the same experimental conditions, DT95% of inoculated Azalea plants decreased remarkably to about 27 h. Subsequent additions of toluene further increased the removal efficiency of the bioaugmented system (DT95% decreased by a factor of four). A decrease in DT95% was also recorded after repeated incubations of non-inoculated plants, but the toluene-removal rate was remarkably low, compared with the inoculated plants. Hence, inoculation of the leaf surface appeared essential for obtaining rapid removal rates. It was not possible to obtain comparable and sustained removal of airborne toluene by inoculating artificial plant surfaces. This is, to our knowledge, the first report on bioaugmentation of the leaf surface of plants to remove gaseous pollutants from air. The results presented are promising and could be of great practical importance in the field of indoor air pollution control. PMID- 12910330 TI - Diffusion of new medicines into hospital practice. PMID- 12910329 TI - Synthesis of FimH receptor-active manno-oligosaccharides by reverse hydrolysis using alpha-mannosidases from Penicillium citrinum, Aspergillus phoenicis and almond. AB - Recombinant Penicillium citrinum alpha-1,2-mannosidase, expressed in Aspergillus oryzae, was employed to carry out regioselective synthesis of alpha- d mannopyranosyl-(1-->2)- d-mannose. Yields (w/w) of 16.68% disaccharide, 3.07% trisaccharide and 0.48% tetrasaccharide were obtained, with alpha1-->2 linkages present at 98.5% of the total linkages formed. Non-specific alpha-mannosidase from almond was highly efficient in reverse hydrolysis and oligosaccharide yields of 45-50% were achieved. The products of the almond mannosidase were a mixture of disaccharides (30.75%, w/w), trisaccharides (12.26%, w/w) and tetrasaccharides (1.89%, w/w) with 1-->2, 1-->3 and 1-->6 isomers. alpha-1,2-linkage specific mannosidase from P. citrinum and alpha-1,6-linkage-specific mannosidase from Aspergillus phoenicis were used in combination to hydrolyse the respective linkages from the mixture of isomers, resulting in alpha- d-mannopyranosyl-(1- >3)- d-mannose in 86.4% purity. The synthesised oligosaccharides can potentially inhibit the adhesion of pathogens by acting as "decoys" of receptors of type-1 fimbriae carried by enterobacteria. PMID- 12910332 TI - How do I become a forensic scientist? Educational pathways to forensic science careers. PMID- 12910333 TI - Forensic discrimination of photocopy and printer toners II. Discriminant analysis applied to infrared reflection-absorption spectroscopy. AB - Copy toner samples were analyzed using reflection-absorption infrared microscopy (R-A IR). The grouping of copy toners into distinguishable classes achieved by visual comparison and computer-assisted spectral matching was compared to that achieved by multivariate discriminant analysis. For a data set containing spectra of 430 copy toners, 90% (388/430) of the spectra were initially correctly grouped into the classifications previously established by spectral matching. Three groups of samples that did not classify well contained too few samples to allow reliable classification. Samples from two other pairs of groups were similar and often misclassified. Closer examination of spectra from these groups revealed discriminating features that could be used in separate discriminant analyses to improve classification. For one pair of groups, the classification accuracy improved to 91% (81/89) and 97% (28/29), for the two groups, respectively. The other pair of groups were completely distinguishable from one another. With these additional tests, multivariate discriminant analysis correctly classified 96% of the 430 R-A IR toner spectra into the toner groups found previously by spectral matching. PMID- 12910331 TI - Pharmacokinetics of anti-influenza prodrug oseltamivir in children aged 1-5 years. AB - OBJECTIVE: The prodrug oseltamivir has been shown to be efficacious and safe for the treatment of influenza for patients 1 year of age or older; however, pharmacokinetic information was lacking for children below 5 years of age. This study was conducted to assess the metabolic and excretory capacity of oseltamivir and its active carboxylate metabolite in young children. METHODS: Twelve healthy children aged 1-5 years received a single oral suspension dose of oseltamivir (45 mg for 3-5 years, 30 mg for 1-2 years). Plasma and urine concentrations of oseltamivir and the carboxylate were determined by means of liquid chromatography/tandem mass spectrometry. RESULTS: Mean peak plasma concentration and area under the plasma concentration-time curve values normalized to milligram per kilogram oseltamivir dose in the 1- to 2-year group are lower than those in the 3- to 5-year group. Mean body weight normalized oral clearance of oseltamivir and its carboxylate in younger subjects aged 1-2 years (259 ml/min/kg and 12.2 ml/min/kg) were, respectively, 52% and 30% higher than those in older subjects aged 3-5 years (170 ml/min/kg and 9.4 ml/min/kg). CONCLUSION: The results demonstrate that infants as young as 1 year old can metabolize and excrete oseltamivir efficiently. The data derived from this study provide the starting dose of oseltamivir for further investigation in an efficacy study among influenza-infected infants less than 1 year of age. PMID- 12910334 TI - Rehabilitation of avulsion fracture of the tibial tuberosity following Osgood Schlatter disease. AB - A sixteen-year-old boy suffered from sharp pain in the knee during a jump while playing basketball. He had a positive history of Osgood Schlatter disease. Radiographic evaluation demonstrated an avulsion fracture of the tibial tuberosity Type III according to the classification of Watson-Jones. Rehabilitation after avulsion fracture of the tibial tuberosity is an important consideration for this relatively uncommon adolescent injury. In such avulsion fractures, landing on the ground with the knee fully extended after a jump is the most likely cause. This case report reviews the rehabilitation program, and selected functional outcome measures after rehabilitation are reported. The patient returned to sporting activity after 12 months. PMID- 12910336 TI - An exploratory randomised controlled trial of a support programme for carers of patients with a psychosis. AB - BACKGROUND: Despite an acknowledgement of the impact of serious mental disorders on informal caregivers, we still know little about how to best help them. The aim of the study was to evaluate the effectiveness of a two-phased carers' intervention comprising family sessions followed by relatives' groups. This intervention was designed to be of 'intermediate' intensity, that is,one lying between brief educational programmes and long-term family psychoeducational treatments. METHODS: An exploratory randomised controlled trial was conducted comparing the experimental support programme with 'standard' care. All carers of patients with a psychotic disorder from a defined population were approached. Outcome measures were based on a 'stress-appraisal-coping' model of caregiving. RESULTS: Despite concerted attempts to engage carers, only 42% participated in the study. The carers' programme did not offer any significant advantage on any of the outcome measures: psychological morbidity, negative appraisal, coping or social support. The severity of caregiving difficulties decreased over the study period for the group as a whole. CONCLUSIONS: There is still uncertainty about the most effective interventions for carers. Meeting 'needs' may not improve caregiver distress. PMID- 12910337 TI - Risk factors for male and female suicide decedents ages 15-64 in the United States. Results from the 1993 National Mortality Followback Survey. AB - BACKGROUND: Few controlled studies have examined possible gender differences in risk factors for suicide. This paper examined the associations of certain risk factors with suicide among males and females aged 15-64, and the variation in the associations by gender. METHODS: A case-control study was constructed from the 1993 National Mortality Followback Survey in the United States. Information concerning age, race, education, living arrangement, marijuana use, excessive alcohol consumption, access to a firearm, depressive symptoms, and mental health service utilization was collected via death certificate and proxy respondent. Decedents between the ages of 15 and 64 who died by suicide were compared with those who died of natural causes. Logistic regression analysis was used to examine the associations between risk factors and suicide in males and females. RESULTS: In comparison to those who died of natural causes, we found that marijuana use, excessive alcohol consumption, and access to a firearm increased the odds of suicide for both genders. For male decedents, the presence of depressive symptoms was more frequently reported for the suicide decedents in the 45-64 age group, and the proportion of mental health service use was higher among suicide decedents who did not complete high school. For female decedents, depressive symptoms were related to suicide in all age groups, and the use of mental health services was more frequent in the suicides of the 15-29 and 45-64 age groups. CONCLUSIONS: The risk factors of marijuana use, excessive alcohol use, and firearm accessibility in the last year of life increased the odds of suicide in both genders. When compared to natural deaths, depressive symptomatology was common in female suicide decedents, whereas it was only associated with older age among male suicide decedents. The interactions of mental health service use with demographic factors suggested possible gender differences in suicide risk associated with severity of mental disorders, as well as the likelihood of treatment seeking. PMID- 12910335 TI - Right ventricular function and positive pressure ventilation in clinical practice: from hemodynamic subsets to respirator settings. PMID- 12910338 TI - The role of managed care and financing in medical practices: how does psychiatry differ from other medical fields? AB - BACKGROUND: The organization and financing of medical practice has experienced substantial changes during the past decade. Today the majority of individuals with private insurance are enrolled in managed care plans and traditional indemnity insurance is only predominant among Medicare enrollees. At the same time that managed care was growing in general, there was also substantial growth in the number of managed care companies specializing in behavioral health care and separating them out ("carved-out") from other medical care. While it is clear that these changes in managed care penetration and financing arrangements have affected physicians and the way they practice medicine, it is less clear whether there has been a differential effect across physician types. METHOD: Data are from the Community Tracking Study (CTS) physician survey, a national survey of active physicians in the United States fielded between August 1998 and November 1999. To be eligible, physicians had to have completed their medical training (which excludes residents, interns, or fellows), be practicing in the contiguous United States, and be providing direct patient care for at least 20h per week. The total number of completed interviews was 12,304, including 566 psychiatrists, 7,217 primary care physicians, and 4,521 other medical or surgical specialists. Consistent with the previous literature, our analysis is primarily descriptive presenting weighted means and odds ratios from multivariate models. RESULTS: Psychiatrists are significantly less likely than primary care physicians or other specialists to agree with the statements that clinical decisions in the interest of their patients do not reduce income. Further, in comparison to primary care physicians, psychiatrists are less likely to agree that it is possible to maintain continuing relationships with patients over time that promote the delivery of high quality care and that they have the freedom to make clinical decisions that meet their patients' needs. In contrast to the perceived impacts on practice, psychiatrists tend to work in practices with a lower dependence on managed care than either other specialists or primary care physicians. Regarding individual physician compensation among non-owners, psychiatrists are less likely than other physicians to have their income adjusted based on individual productivity. CONCLUSIONS: Psychiatrists are more concerned that managed care has a negative impact on patient-provider relationships than other physicians; however, based on the available measures of financing and organization, psychiatrists are less dependent on managed care. This may suggest that other aspects of managed care, such as pre-authorization and gatekeeping, that disproportionately affect specialists are driving psychiatrists' perceptions of the patient-provider relationship. PMID- 12910340 TI - Stress, social support and depression in single and married mothers. AB - BACKGROUND: This study examined the effect of stress and social support on the relationship between single-parent status and depression. METHOD: A secondary data analysis of the 1994-95 National Population Health Survey was conducted. Single and married mothers who participated in the survey were derived from the general sample (N = 2,921). Logistic regression techniques were used to assess the mediating and moderating effects of stress and social support on the relationship between family structure and depression. RESULTS: Bivariate analyses showed that, compared to married mothers, single mothers were more likely to have suffered an episode of depression (12-month prevalence), to report higher levels of chronic stress, more recent life events and a greater number of childhood adversities. Single mothers also reported lower levels of perceived social support, social involvement and frequency of contact with friends and family than married mothers. The results of the multivariate analyses showed that, together, stress and social support account for almost 40% of the relationship between single- parent status and depression. We also found a conditional effect of stress on depression by family structure. Life events were more strongly related to depression in married than in single mothers. CONCLUSIONS: A substantial part of the association between single-parent status and depression can be accounted for by differences in exposure to stress and social support. Our results suggest that it is important to examine multiple sources of stress, as exposure to both distal and proximal stressors were higher among single mothers. Limitations and directions for future research are discussed. PMID- 12910339 TI - Life events are important in the course of hopelessness-a 2-year follow-up study in a general population. AB - BACKGROUND: The association between life events and hopelessness in a general population is unknown. AIM: The aim of this study was to examine the course of hopelessness and how positive and negative life events are associated with it. METHOD: This was a 2- year follow-up study among general population adults, excluding any with a mental disorder. The impact of 15 occasional life events during the follow-up was assessed and the course of hopelessness measured with the Beck Hopelessness Scale (HS). RESULTS: Four percent of the study subjects with no hopelessness at baseline and 56% of those with hopelessness at baseline reported hopelessness on follow-up. In multiple logistic regression analyses, a notable worsening of the subjective financial situation was revealed as the most important life event, both in becoming hopeless during the follow-up (OR 5.07; 95% CI 2.20-11.7) and in continued hopelessness (OR 7.51, 95% CI 2.19-25.8). Moreover, considerable interpersonal conflicts at work (OR 3.29, 95% CI 1.17 9.27) were associated with becoming hopeless. However, a notable positive change in common living conditions (OR 0.16, 95% CI 0.04-0.74) was found to be a protective factor against becoming hopeless. All these variables remained significant even when adjusted for change in depression scores (BDI). CONCLUSION: Hopelessness may be persistent in a general population. The impact of life events, especially a notable worsening of the subjective financial situation, is important in becoming or remaining hopeless. PMID- 12910341 TI - Predictors of social services supervision of babies of mothers with mental illness after admission to a psychiatric mother and baby unit. AB - BACKGROUND: There is insufficient information on the predictors of parenting difficulties in mothers with severe mental illness. Using data from mother and baby units in the UK we aimed to examine the social and clinical characteristics of mothers whose babies were supervised by social services on discharge. METHOD: A case-control study was carried out using data from mother and baby units and facilities entered onto the Marce database. RESULTS: Of 1197 mothers, 23% were discharged with their babies under some form of social services supervision. Factors independently associated with an increased risk of supervision included social class (OR 3.16, 95% CI 1.99-5.03), single marital status (OR 2.10, 95% CI 1.38-3.20), behavioural disturbance (OR 1.69, 95% CI 1.08-2.65) and psychiatric illness in the partner (OR 2.67, 95% CI 1.59-4.49). The diagnostic groups independently associated with the highest risk of having a supervised baby were schizophrenia (OR 5.16, 95% CI 2.61-10.21) and personality disorder (OR 9.29, 95% CI 3.46-24.91). CONCLUSIONS: Mothers with schizophrenia are at particularly high risk of having their baby supervised by social services. Preventative interventions should be targeted at socio-economic difficulties, early detection of psychiatric disorders postpartum and treatment of perinatal mental illness in the context of the whole family. PMID- 12910342 TI - The aetiology of posttraumatic stress disorder in four ethnic groups in outpatient psychiatry. AB - BACKGROUND: PTSD is one of few diagnoses to be defined by its aetiology. At treatment centres specialising in a certain type of trauma, like war, torture or sexual abuse, the aetiology may be regarded as self-explanatory. Recent surveys of general populations reveal high rates of PTSD, often following exposure to multiple traumatic events. Traumas are frequently overlooked in general psychiatric settings and PTSD is underestimated. Such findings indicate the need to conceptualise trauma in terms of multiple events and to examine the aetiology of PTSD amongst patients in everyday psychiatric practice. The present study examines aetiological factors of PTSD in three minority groups at a general psychiatric outpatient clinic. Arabs, Iranians and Turks selected from an entire year's cohort of patients-and a sample of Swedish patients of similar age and gender distribution. METHODS: Histories were collected from 115 patients, both regarding their own traumas and traumas involving relatives and close friends, by means of a questionnaire. A self-rating instrument for PTSD, with no reference to a specific trauma, was used in order to study the relationship between trauma and PTSD outcome. RESULTS: Of the patients, 89% had experienced at least one trauma, and 77% multiple traumatic events. The prevalence of probable PTSD varied between ethnic groups: Iranians 69%, Arabs 59%, Turks 53% and Swedes 29%. In a logistic regression analysis, probable PTSD outcome was associated with multiplicity of relatives' traumas (OR = 3.14), multiplicity of own traumas (OR = 2.56) and belonging to an ethnic minority (OR = 2.44), but not with gender. No patient without a trauma history was positively assessed for probable PTSD. CONCLUSION: The symptomatic syndrome of probable PTSD does not appear in the absence of extreme trauma, lending empirical support to the definition of PTSD. Prevalence increases with multiplicity of traumas, including those involving a relative or close friend. The high prevalence of probable PTSD found in this study warrants screening for trauma among patients, their relatives and close friends, in outpatient psychiatry. PMID- 12910343 TI - Does racial bias influence psychiatric diagnoses assigned at first hospitalization? AB - BACKGROUND: Research on racial bias in psychiatric diagnosis has largely been limited to studies of admission diagnoses assigned to chronically ill patients. This study tests whether racial bias influences diagnoses assigned to patients at discharge from their first psychiatric hospitalization. METHODS: In a county-wide sample of patients with psychosis, hospital diagnoses were compared with research diagnoses formulated using structured interviews and strict adherence to DSM-III R. Symptom patterns were also examined. RESULTS: Racial differences were observed in the distribution of both hospital and research diagnoses. Using research diagnoses as the gold standard, the sensitivities and specificities of hospital diagnoses were similar by race (for blacks the sensitivity and specificity of schizophrenia was 0.33 and 0.91, and for whites, 0.43 and 0.89). The only suggestion of possible bias was that more blacks were discharged without a definitive diagnosis (38.7% of blacks vs. 26.3% of whites, chi(2) = 5.80, df = 1, p = 0.02). CONCLUSIONS: We did not observe the expected racial bias in the assignment of diagnoses of schizophrenia and affective disorders. While there was evidence that hospital clinicians had more difficulty diagnosing black patients, the low concordance between hospital and research diagnoses for both black and white patients demonstrates the need to better understand the clinical diagnostic process for all patients with psychotic disorders at their first hospitalization. PMID- 12910344 TI - Genetic mapping of QTLs affecting productivity and plant architecture in a full sib cross from non-inbred parents in Cassava (Manihot esculenta Crantz). AB - An attempt was made to identify quantitative trait loci (QTLs) for several productivity and plant architecture traits in a full-sib progeny of 144 individuals from two non-inbred parents in cassava. A molecular linkage map of this cross constructed previously with over 250 markers was the source of molecular markers. The progeny were grown under field conditions at two locations (Palmira and Quilichao) in Colombia and evaluated in 2 years (1998 and 1999) for architecture and productivity traits. Architecture traits evaluated were plant height (PH), branching height (BH), branching levels (BL), branching index (BI), stem portion with leaves (SPL) and leaf area index (LAI). Productivity traits were those related to total dry matter production and distribution, namely fresh root yield (FRY), fresh shoot yield (FSY), harvest index (HI) and the number of storage roots (NR). Phenotypic evaluation of the traits in this population revealed continuous variation for all traits. Broad-sense heritability estimates, ranged from 36% (for NR) to 94% (for BH). Several significant phenotypic correlations were observed between architecture and productivity traits. Primary QTLs, using the single-QTL model, and secondary QTLs, by a primary QTL interaction model, were detected by interval mapping. A total of 30 primary QTLs and 84 secondary QTLs were detected. We identified 35% of detected QTLs in two or more trials, the other QTLs were environment-specific. These results underscore the significant genotype x environment interactions found for most of the traits. Several genomic segments affecting multiple traits were identified and were in agreement with correlation among traits. All QTLs identified for FRY were found associated with either component traits of productivity or architecture traits. This study suggests that QTLs for plant architecture can be used to improve productivity. However an exhaustive search and analysis of QTLs controlling architecture is required before marker-assisted selection (MAS) for increasing productivity can be initiated. PMID- 12910345 TI - Factors limiting maximal performance in humans. AB - Theoretical best performance times (ttheor) in track running are calculated as follows. Maximal metabolic power (Emax) is a known function of maximal oxygen uptake (VO2max), of maximal anaerobic capacity (AnS) and of effort duration to exhaustion (te):Emax=f (te). Metabolic power requirement (Er) to cover the distance (d) in the performance time tp is the product of the energy cost of locomotion per unit distance (C) and the speed: Er=Cxd/tp. The time values for which Emax (te)=Er (tp), assumed to yield ttheor, can be obtained for any given subject and distance provided that VO2max, AnS and C are known, and compared with actual best performances (tact). For 15 min> or =te> or =100 s, the overall ratio tact/ttheor was rather close to 1.0. To estimate the relative role of the different factors limiting VO2max, several resistances to O2 transport are identified, inversely proportional to: alveolar ventilation (RV*), O2 transport by the circulation (RQ), O2 diffusion from capillary blood to mitochondria (Rt), mitochondrial capacity (Rm). Observed changes of VO2max are accompanied by measured changes of several resistances. The ratio of each resistance to the overall resistance can therefore be calculated by means of the O2 conductance equation. In exercise with large muscle groups (two legs), RQ is the major (75%) limiting factor downstream of the lung, its role being reduced to 50% during exercise with small muscle groups (one leg). Rt and Rm account for the remaining fractions. In normoxia RV* is negligible; at high altitude it increases progressively, together with Rt and Rm, at the expense of RQ. PMID- 12910347 TI - Features of acute and transient psychotic disorders. AB - BACKGROUND: Little is known about acute and transient psychotic disorders (ATPD), a diagnostic category introduced with ICD-10. AIMS: To determine the clinical and sociodemographic features, course and outcome of ICD-10 ATPD in a prospective and longitudinal study. METHOD: We recruited all consecutive inpatients fulfilling the ICD-10 criteria of ATPD during a 5-year period. Demographic and clinical features were systematically evaluated and follow-up investigations were carried out at an average of 10 years after onset of the disorder using standardized instruments. RESULTS: ATPD patients represented 8.5% of all inpatients with non organic psychotic disorders. ATPD were characterized by female preponderance. In two-thirds of the cases a typical polymorphic symptomatology was found. In spite of the fact that the possibility of relapse within 5 years was high, the psychopathological and social outcome for most of the patients was very favourable. Schizophrenic episodes during follow-up were rare (7.7% of patients), but a strictly monomorphous course (ATPD episodes only) from index episode to the end of the prospective follow-up was found in only 53.9% of the patients. CONCLUSION: ATPD are not a sharply demarcated and unchanging nosological entity. Nevertheless, the present data support a delineation of ATPD as a diagnostic category with specific clinical features and with a usually favourable prognosis. Further research on the topic is necessary. PMID- 12910349 TI - Self-rated aggression and cerebral monoaminergic turnover. Sex differences in patients with persistent depressive disorder. AB - OBJECTIVE: Outward-directed violence and impulsivity in humans and primates has frequently been related to abnormal brain monoaminergic turnover. Self-rated aggression is likely to be clinically relevant,and its psychobiological basis needs investigation. SUBJECTS: Sixty-six patients (40 women and 26 men) with persistent depressive disorder (PDD) were compared with 497 control subjects from the general Swedish population. METHODS: We administered the Aggression Questionnaire - Revised Swedish Version (AQ-RSV) to patients and control subjects. In patients, CSF 5-hydroxyindoleacetic acid (5-HIAA) and 3-methoxy-5 hydroxyphenylglycol (MHPG) in CSF were analyzed. Total Aggression score and Aggression subfactors 'Physical Aggression','Verbal Aggression','Anger', and 'Hostility'were correlated with CSF concentrations of 5-hydroxyindoleacetic acid (5-HIAA),and 3-methoxy-5-hydroxyphenylglycol (MHPG). RESULTS: Overall, Hostility was positively related to CSF MHPG (t=2.27, p=0.015). Split by sex,Hostility was related with 5-HIAA in males (r=0.62,p=0.003),and with MHPG in females (r=0.38, p=0.03). Comparing self rated aggression with age- and sex-matched data from the general Swedish population, the most prominent deviation was increased Hostility score among PDD patients. Among patients, all aggression factors were nominally higher in women than in men, with the most pronounced sex difference in Hostility (t=-1.89, p=0.04). CONCLUSIONS: Results suggest a clinically meaningful sex difference in a positive relationship between hostility and serotonergic/noradrenergic turnover in PDD patients. PMID- 12910348 TI - Atypical and typical neuroleptics in acute schizophrenia and related delusional disorders. Drug choice, switching and outcome under naturalistic treatment conditions. AB - Atypical neuroleptics have improved drug treatment in schizophrenia. However, their use varies greatly between countries and continents. Recent metaanalyses have deemphasized the range and magnitude of their superiority compared to typical neuroleptics. Aims of the present study were to contribute effectiveness data to this discussion. In 725 inpatients with ICD-10 diagnoses F20, 22-25 from four German psychiatric inpatient units acute neuroleptic treatment and outcome were analyzed under naturalistic conditions. Treatment strategies were stratified post hoc to answer the question, which proportion - and which kind - of patients are primarily given atypicals or typicals, for how long, at which rate and when the atypical/typical drugs are switched to typical/atypical drugs, and what the respective outcomes are. As the results demonstrate, atypicals were administered one time during inpatient treatment in nearly 48% of the patients, however as first choice drugs in only 15% of this population. Treatment change occurred in 28% after 5-6 weeks irrespective of the first drug choice. Outcome differences were, if at all, only modest and not systematically biased towards a single strategy. In conclusion, frequency of inpatient treatment with atypical neuroleptics corresponds to pharmaco-epidemiological data in Europe, but is still lower than in the US. Contrary to contemporary guideline recommendations atypical neuroleptics under routine inpatient treatment conditions were scarcely administered as first choice treatment, and acute clinical outcome is comparable to that under treatment with typical neuroleptics. Reasons and implications of these findings considering the methodological limitations are discussed. PMID- 12910346 TI - Interrelations between monoaminergic afferents and corticotropin-releasing factor immunoreactive neurons in the rat central amygdaloid nucleus: ultrastructural evidence for dopaminergic control of amygdaloid stress systems. AB - Ample evidence implicates corticotropin-releasing factor (CRF)-producing neurons of the central amygdaloid nucleus (CeA) in vegetative, endocrine, and behavioral responses to stress and anxiety in laboratory rats. Monoaminergic systems are involved in modulating these responses. In the present paper, interrelations between CRF-immunoreactive (ir) neurons, and noradrenergic, serotonergic, and dopaminergic afferents were studied using single and double immunolabeling for light and electron microscopy in the rat CeA. Dopaminergic axons formed dense plexus in the CeA overlapping with the localization of CRF-ir neurons, and their terminals formed frequent associations with CRF-ir somata. Contacts of serotonergic axons on CRF-ir neurons were few, and contacts of noradrenergic axons were the exception. Ultrastructurally, symmetric synapses of dopaminergic terminals on CRF-ir somata and dendrites were found. More than 83% of CRF-ir somata were contacted in single ultrathin sections. About half of these possessed two or more contacts. Of non-ir somata, 37% were contacted by dopaminergic terminals, and only 13% of these had two or more contacts. Correlative in situ hybridization indicated that CeA CRF-ir neurons may express receptor subtype dopamine receptor subtype 2. In conclusion, dopaminergic afferents appear to specifically target CeA CRF neurons. They are thus in a position to exert significant influence on the rat amygdaloid CRF stress system. PMID- 12910350 TI - Gonadal hormones in schizophrenia and mood disorders. AB - There are gender-related differences in the prevalence, course and treatment response characteristics of schizophrenia and mood disorders. Gonadal steroids exert potent effects on mood, cognition and behavior, and there is little doubt that androgens are crucial for differentiating to each gender. Serum level of total testosterone, free testosterone, estradiol and sex hormone binding globulin was measured in 69 medication-free men with either schizophrenia (n=29) or bipolar I disorder, manic episode (n=18) or major depressive disorder (n=22). There was a statistically significant difference in free testosterone level between mania and schizophrenia groups (p<0.05). The higher free testosterone level in the mania group compared to the schizophrenia group found in this study supports further investigation of a potential difference in the hypothalamic pituitary-gonadal axis between patients with schizophrenia and bipolar I disorder, manic. PMID- 12910351 TI - Dimensions of suicidal behavior according to patient reports. AB - Five factor analyses with limitations explored the Suicidal Intent Scale (SIS) subscales reflecting suicidal behavior dimensions. This larger sample study conducts an exploratory factor analysis of the SIS. Two large samples of suicide attempters (N= 435 and N= 252) from a general hospital were studied. The validity of SIS subscales obtained from the factor analysis was investigated by examining the association between the subscales and clinical variables. There were two factors: expected lethality and planning. In both samples, male gender and depression tended to be associated with higher scores in both subscales (small to medium effect sizes). Hospitalization was associated with higher scores in both SIS subscales (medium to large effects) suggesting that these subscales were reasonably good predictors of suicide attempt severity. Clinicians assessing patient reports to establish the severity of suicide attempts need to ask questions regarding both dimensions: expected lethality and planning. PMID- 12910352 TI - Frequency of bipolar spectrum in 111 private practice depression outpatients. AB - BACKGROUND: Mood disorders included into the bipolar spectrum are increasing, and overactivity (increased goal-directed activity) has reached the status of mood change for the diagnosis of hypomania in the recent studies by Angst and Akiskal. STUDY AIM: was to find frequency of bipolar spectrum in remitted depressed outpatients by including sub-syndromal hypomania. METHODS: 111 depression remitted outpatients were interviewed for history of hypomania and hypomanic symptoms with the Structured Clinical Interview for DSM-IV-Clinician Version (a partly semistructured interview), as modified by Benazzi and Akiskal. Bipolar I patients were not included. All past hypomanic symptoms (especially overactivity) were systematically assessed. Wording of the questions could be changed to increase/check understanding. Subsyndromal hypomania was defined as an episode of overactivity (increased goal-directed activity) plus at least 2 hypomanic symptoms. RESULTS: Frequency of bipolar II (BPII) was 68/111 (61.2%, 95% confidence interval 52% to 69.8 %), frequency of major depressive disorder (MDD) was 43/111. The most common hypomanic symptom was overactivity. In the MDD sample, sub-syndromal hypomania was present in 39.5% (15.3% of the entire sample), and had 4 median symptoms. Bipolar spectrum frequency was 76.5% (95% confidence interval 67.9% to 83.5 %). Overactivity had higher sensitivity than elevated mood for predicting BPII diagnosis. LIMITATIONS: Single interviewer. CONCLUSIONS: By systematic probing more focused on past overactivity than mood change, and by inclusion of sub-syndromal hypomania, bipolar spectrum frequency was higher than the near 1 to 1 ratio versus MDD reported up to now (Angst et al.). Given the wide confidence interval, the value in the depression population should be around 70%. Better probing skills by clinicians, and use of semi structured interviews could much reduce the current high underdiagnosis of BPII and related disorders in usual clinical practice. PMID- 12910353 TI - Course and outcome of first-admitted patients with acute and transient psychotic disorders (ICD-10:F23). Focus on relapses and social adjustment. AB - OBJECTIVE: The aim of the present study was to investigate course and outcome of acute and transient psychotic disorders (ATPD). METHOD: A sample of 73 first hospitalized patients was evaluated after three to seven years in order to determine the frequency of relapses and to assess social adjustment. RESULT: Forty-two percent experienced no relapse, 46% experienced relapses without developing marked deficits in social adjustment and 12% had relapses associated with a severe social impairment. At discharge from first hospitalization the last group was distinguishable from the other two with respect to negative and depressive symptoms as well as the total score of the Strauss-Carpenter scale. CONCLUSION: Only a minority of first-hospitalized patients with ATPD develop a severe social impairment after three to seven years. This subgroup, however, is not compatible with the concept of a "transient" psychotic disturbance, but rather with an early manifestation of a chronic schizophrenic disorder. PMID- 12910354 TI - Is executive function associated with symptom severity in schizophrenia? AB - This paper reports on a study that was designed to investigate the relationship between psychopathology and executive functions in schizophrenia. Correlations were sparse and mostly weak. The most robust finding was the association between letter fluency and negative symptoms; however, most other applied tasks were not associated with symptom level. Our results support previous findings of differential relationships between impaired executive functions and symptom level. PMID- 12910356 TI - Tape stripping and sodium dodecyl sulfate treatment increase the molecular weight cutoff of polyethylene glycol penetration across murine skin. AB - Prior studies in hairless mice have demonstrated that acute barrier disruption by acetone treatment increases the molecular weight (MW) cutoff of polyethylene glycol (PEG) penetration through the skin. The objective of the present study was to further investigate the dependence of permeability on MW with different forms of barrier disruption. A series of PEGs ranging in MW from near 300 to over 1000 Da were used to study the effects of tape stripping and sodium dodecyl sulfate (SDS) treatment on the MW permeability profiles of mouse skin in vitro. The 12-h percutaneous penetration of all the PEG 300, 600, and 1000 oligomers generally increased as a function of transepidermal water loss (TEWL) of the skin, either tape-stripped or SDS-treated. In addition, the total penetration of PEG oligomers across control skin, and skin tape-stripped and SDS-treated to different degrees of barrier disruption progressively decreased with increasing MW. There were no significant differences in the percutaneous penetration of the PEG oligomers between skin tape-stripped and SDS-treated to the same degree of barrier disruption. The penetration enhancement relative to control skin was more prominent with larger molecules. The MW cutoff for skin penetration increased with the degree of barrier disruption irrespective of the treatment applied, and was 986 Da (tape stripping) and 766 Da (SDS treatment) at TEWL levels in the range 10-20 g/m(2) per h in comparison with 414 Da for control skin. In accordance with previous findings in acetone-treated mouse skin, the results strongly suggest that, irrespective of the form of barrier disruption applied, not only higher amounts but also more varieties of chemicals (larger molecules) may penetrate skin with a compromised barrier than normal skin. PMID- 12910357 TI - Patellar hypertension syndrome in adolescence: a three-year follow up. AB - INTRODUCTION: Anterior knee pain is one of the major causes of knee pain in an adolescent population. It has been suggested that raised intraosseous pressure may play an important role in the development of anterior knee pain in the young patient. MATERIAL AND METHODS: A prospective clinical multicentre study with 27 adolescent patients presenting with anterior knee pain was performed. After failure to respond to conservative treatment, all patients agreed to undergo minimally invasive intraosseous pressure measurements in local anaesthesia. All 27 patients demonstrated a positive "pain provocation test" as a response to increased intrapatellar pressure and were decompressed with a new intraosseous drilling technique. RESULTS: The repeat intraosseous pressure measurement in 27 patients 1 year postoperatively revealed a significant reduction from 43.3+/-13.4 preoperatively to 24.1+/-4.2 mmHg (p<0.001). The pain relief was documented in the form of a reduced VAS from 7.6+/-1.4 to 2.1+/-1.1 cm after 3 years (p<0.001). CONCLUSION: Patients with anterior knee pain and a positive "pain provocation test" were diagnosed as suffering from a hypertension syndrome of the patella. We conclude that simple extraarticular patellar decompression may offer a valuable new form of treatment in this select group of adolescent patients with anterior knee pain. PMID- 12910361 TI - Primary anastomosis after intraoperative colonic lavage vs. Hartmann's procedure in generalized peritonitis complicating diverticular disease of the colon. AB - BACKGROUND AND AIMS: For complicated diverticulitis Hartmann's procedure remains the favored option in patients with acute complicated sigmoid disease, but there has been increasing interest in primary resection and anastomosis with intraoperative colonic lavage. This study compared primary resection with intraoperative colonic lavage and Hartmann's procedure. PATIENTS AND METHODS: Between January 1994 and November 2001, 60 patients underwent emergency laparotomy for diverticular peritonitis (Hinchey stages III and IV). Primary resection and anastomosis with intraoperative colonic lavage was performed in 27 patients and Hartmann's procedure in 33. All data were collected prospectively on a standardized form. RESULTS: Mortality with intraoperative colonic lavage was 11% and with Hartmann's procedure 12%. The incidence of postoperative complication was significantly higher after Hartmann's procedure. The mean hospital stay was significantly longer after Hartmann's procedure than after primary resection with intraoperative colic lavage. CONCLUSION: Primary resection with intraoperative colonic lavage compares favorably with Hartmann's procedure for diffuse purulent peritonitis in complicated diverticulitis. It should be an alternative to Hartmann's procedure in stercoral peritonitis. PMID- 12910360 TI - MELAS: clinical phenotype and morphological brain abnormalities. AB - We describe the clinical and neuropathological findings of three unrelated autopsy cases of MELAS harboring the A3243G transition in the mitochondrial DNA (mtDNA). Using immunohistochemical techniques, we studied the expression of several subunits of the respiratory chain in various brain regions from the same cases. In all three cases there was a reduced immunocytochemical staining for mtDNA-encoded subunits of the respiratory chain, confirming the presence of a defective mitochondrial protein synthesis in this disease. Mitochondrial abnormalities were mostly confined to multiple areas of different size and shape, in agreement with the focal character of the brain pathology in MELAS, and were most prominent in the cerebral cortex, providing a morphological contribution to the explanation of the cognitive regression of the patients. Immunoreactivity for mtDNA-encoded subunits was reduced in the walls of many pial and intracerebral arterioles of different brain regions but there was no clear correlation between territories of affected vessels and distribution of the histological and immunohistochemical lesions. Cerebral focal lesions in MELAS might have a metabolic nature and several pathogenetic mechanisms might be involved in the genesis of stroke-like episodes when there is a local increased ATP demand. PMID- 12910362 TI - Sugar preferences and feeding strategies in the hawkmoth Macroglossum stellatarum. AB - Hummingbird hawkmoths were tested for their preferences for different types of sugar. In triple choice tests, moths sucked for longer periods from sucrose than from fructose and glucose. Naive moths released in a large flight cage and monitored over 24 days, drank on average 137 mul sucrose, 67 mul fructose and 7 mul glucose daily. In an independent test, moths spent more time feeding from sucrose than from fructose and more time feeding from fructose than from glucose. Animals in hibernation made less but longer feeding bouts, whereas animals that fed every day and newly eclosed animals were more likely to make more but shorter visits to feeders. The hawkmoths learned to associate colour with the preferred sugar. In a dual choice test, flower-naive moths chose blue rather than yellow artificial flowers. After the initial test, these animals received sucrose from the yellow and glucose from the blue feeders. A week later they chose yellow more frequently than blue indicating that they had learned to associate a colour with the preferred type of sugar. PMID- 12910363 TI - External beam radiation therapy: role of androgen deprivation. PMID- 12910364 TI - Radiotherapeutic approaches to metastatic disease. AB - Bone metastases in patients with prostate cancer are common. Prostate cancer patients also have a relatively long survival. Palliative radiation can provide effective symptom control in many clinical settings. This article focuses on the management of bone pain and its complications (pathological fractures and spinal cord compression) with radiation treatment. PMID- 12910366 TI - A tool for understanding homologous recombination in plants. AB - Attempts for establishing an efficient gene targeting (GT) system in seed plants have hitherto not been successful. In contrast, GT based on homologous recombination is highly efficient in Physcomitrella, making this moss a novel tool in reverse genetics. However, why homologous and illegitimate recombination are differently regulated between Physcomitrella and seed plants is still enigmatic. Here we update the state of the art of GT in Physcomitrella and discuss approaches to unravel this enigma. Identification of molecular factors significantly enhancing GT and their subsequent transfer to crop plants will have a great impact on plant biotechnology by enabling precise genetic engineering. Physcomitrella appears to be the most useful model system in this context. PMID- 12910367 TI - Efficient in vitro direct shoot organogenesis and regeneration of fertile plants from embryo explants of Bambara groundnut ( Vigna subterranea L. Verdc.). AB - An efficient protocol has been developed for direct shoot organogenesis from embryo axes derived from mature seeds of two different landraces of Bambara groundnut. Multiple shoots were initiated on several media containing different concentrations and combinations of benzylaminopurine (BAP) or thidiazuron (TDZ). Efficient regeneration occurred when the embryo axes were first plated for 6 days on a medium containing high concentrations of BAP (1 mg/l) and alpha naphthaleneacetic acid (NAA, 1 mg/l) and then cut transversely and transferred onto a medium containing 1.5 mg/l BAP. Shoot regeneration frequency was 100% and from five to eight shoots per explant were obtained. The importance of using embryo explants and cytokinins in the culture media, with respect to controlling the development of a highly organogenic system, was demonstrated. Histological studies revealed that proliferating buds originated directly from the superficial layers of the explants without an intermediate callus phase. The regenerated shoots were rooted on a medium containing 1 mg/l NAA and then transferred to the greenhouse. Flow cytometric analyses and chloroplast counts of guard cells suggested that the regenerants were diploid. All were morphologically normal and fertile. The short duration, high efficiency and low frequency of somaclonal variation of this system make it well suited for wider biotechnological applications of Bambara groundnut-a neglected and under-utilized crop. PMID- 12910365 TI - Molecular and genetic prognostic factors of prostate cancer. AB - Prostate cancer is the most commonly diagnosed cancer in Western males, responsible for 3% of all deaths in men over 55 years of age and second only to lung cancer as a cause of cancer death. Biomarkers have become an important diagnostic tool in prostate cancer. The discovery of the serum marker prostate specific antigen (PSA) significantly facilitated the detection and management of prostate cancer. As we enter into the post-genomics era, novel biomarkers of prostate cancer of therapeutic significance will invariably emerge. Here we review a series of existing and emerging molecular-based prognostic markers particularly with radiotherapy. PMID- 12910368 TI - The impact of Gelrite and activated carbon on the elemental composition of two conifer embryogenic tissue initiation media. AB - The elemental composition of plant tissue culture media was studied in response to (1) different levels of Gelrite and activated carbon (AC) in semisolid media and (2) different levels and types of AC in liquid media. Doubling the level of Gelrite from 2 g/l to 4 g/l reduced available magnesium (20%), calcium (16%), zinc (17%) and manganese (24%) and increased potassium (6%). AC adsorbed copper (90-95%) and zinc (35-51%) from both liquid and semisolid media. Two significantly different ACs gave minor differences in adsorption. No adsorption was indicated for inorganic anions. Nonacid-washed AC released significant levels of magnesium (44% increase), calcium (16% increase) and silica (a 75% increase to 1.8 mg/l). The elemental composition of media may need to be adjusted when increasing the Gelrite level or adding AC. PMID- 12910369 TI - Genetic transformation of the figwort, Scrophularia buergeriana Miq., an Oriental medicinal plant. AB - Scrophularia buergeriana Miq. (figwort) contains a diverse group of bioactive natural products and is used to treat a variety of ailments, including fever, constipation, neuritis, and laryngitis. A transformation protocol was established for S. buergeriana using Agrobacterium tumefaciens. Kanamycin-resistant plants were regenerated from leaf explants co-cultivated with A. tumefaciens strain GV3101. The shoot regeneration medium was supplemented with 2 mg l(-1) 6 benzylaminopurine and 70 mg l(-1) putrescine to improve the efficiency of organogenesis. Detection of the neomycin phosphotransferase gene, the presence of high levels of beta-glucuronidase (GUS) transcripts and enzyme activity, and the histochemical localization of GUS confirmed the genetic transformation of S. buergeriana. This work demonstrates the potential of using A. tumefaciens to efficiently transfer foreign genes into a commercially and culturally important Oriental medicinal plant. PMID- 12910370 TI - A promoter derived from taro bacilliform badnavirus drives strong expression in transgenic banana and tobacco plants. AB - Taro bacilliform virus (TaBV) is a pararetrovirus of the genus Badnavirus which infects the monocotyledonous plant, taro ( Colocasia esculenta). A region of the TaBV genome spanning nucleotides 6,281 to 12 (T1200), including the 3' end of open reading frame 3 (ORF 3) and the intergenic region to the end of the tRNA(met)-binding site, was tested for promoter activity along with four different 5' deletion fragments (T600, T500, T250 and T100). In transient assays, only the T1200, T600, T500 fragments were shown to have promoter activity in taro leaf, banana suspension cells and tobacco callus. When these three promoters were evaluated in stably transformed, in vitro-grown transgenic banana and tobacco plants, all were found to drive near-constitutive expression of either the green fluorescent protein or beta-glucuronidase (GUS) reporter gene in the stem (or pseudostem), leaves and roots, with strongest expression observed in the vascular tissue. In transgenic banana leaves, the T600 promoter directed four-fold greater GUS activity than that of the T1200, T500 and the maize polyubiquitin-1 promoters. In transgenic tobacco leaves, the levels of GUS expression directed by the three promoters was between four- and ten-fold lower than that of the double Cauliflower mosaic virus 35S promoter. These results indicate that the TaBV derived promoters may be useful for the high-level constitutive expression of transgenes in either monocotyledonous or dicotyledonous species. PMID- 12910371 TI - A putative mitochondrial fission gene from the ectomycorrhizal ascomycete Tuber borchii Vittad.: cloning, characterisation and phylogeny. AB - Mitochondrial binary division is a complex process occurring in multiple steps, mediated by several proteins. In Saccharomyces cerevisiae, a mitochondrial membrane protein, Fis1p, is required for the proper assembly of the mitochondrial division apparatus. In this study, we report the cloning, characterisation and phylogenetic analysis of Tbfis1, a gene from the ectomycorrhizal ascomycetous truffle Tuber borchii, encoding for an orthologue of S. cerevisiae Fis1p. The Tbfis1 coding region consists of a 468-nucleotide open reading frame interrupted by four introns, which encodes for a polypeptide of 155 amino acids, having a predicted transmembrane domain structure typical of the Fis1p Family. Southern blot analysis revealed that Tbfis1 is a single-copy gene in the T. borchii genome. Tbfis1 is highly expressed during the first stages of T. borchii fruit body ripening, while its expression decreases during T. borchii mycelium ageing. Also, Virtual Northern blot analysis revealed Tbfis1 expression in the symbiotic phase of the fungus life cycle. Phylogenetic analysis allowed the identification of Tbfis1 orthologues in filamentous fungi, yeasts, plants, worms, flies and mammals, indicating that the function of the protein coded by this gene has been conserved during evolution. PMID- 12910372 TI - Post-transplant interleukin-2 in patients with low-grade lymphoid neoplasms previously treated with fludarabine is limited by hematologic toxicity. AB - Given the favorable immunologic effects of IL-2 post transplant, we conducted a feasibility study examining rIL-2 1.0x106 IU/m2/day (SQ) beginning on D+14 post transplant and continuing for 90 days in 12 patients with low-grade lymphoproliferative disorders. Prior to high-dose chemotherapy and autologous peripheral blood stem cell transplantation (HDCT), 11 patients underwent cytoreduction with fludarabine and cyclophosphamide (Flu/Cy); 11 were in complete remission (CR) and one was in partial remission at the time of HDCT. All 12 patients were in CR 90 days post-HDCT. At a median follow-up of 30 (range 3-44) months, seven patients (58%) remain in remission, four are alive with disease, and one has died of disease progression, resulting in an overall survival of 92%. Kaplan-Meier estimates of progression-free survival (PFS) for the group demonstrate a median of 31 (range 3-43) months. Five patients required rIL-2 cessation at 8-58 days after starting the therapy due to hematologic toxicity. These results are comparable to those achieved in other published bone marrow and peripheral blood stem cell transplantion (PBSCT) series without the addition of rIL-2. Furthermore, rIL-2 using this schedule following fludarabine-based cytoreduction was associated with excessive hematologic toxicity. PMID- 12910373 TI - Reduced-intensity conditioning followed by allogeneic hematopoietic cell transplantation in myeloid diseases. AB - Within the past years, reduced or modified doses of chemotherapy or radiotherapy have been widely studied for conditioning before allogeneic hematopoietic cell transplantation in patients with myeloid leukemia not eligible for conventional transplantation. The main goal was to reduce the substantial treatment-related mortality in this patient population while preserving the potential curative graft-versus-leukemia effect. This review summarizes results of published trials using reduced-intensity conditioning (RIC) in patients with acute myeloid leukemia (AML), myelodysplastic syndrome (MDS), chronic myeloid leukemia (CML), and myelofibrosis. In most of the published trials conditioning contained fludarabine (90-180 mg/m(2)) in combination with busulfan (4 x 10 mg/kg), melphalan (90-140 mg/m(2)), or 2-5 Gy total body irradiation (TBI). Peripheral blood hematopoietic stem cells from related or unrelated donors were used as graft source in most of the studies. Post-transplantation immunosuppression consisted of cyclosporine combined with methotrexate or mycophenolate mofetil. Although the majority of the patients were above the age of 50 years, early treatment-related mortality was rather low. Nevertheless, the rate of clinically significant graft-versus-host disease (GVHD) seemed to be comparable to conventional transplants in most of the protocols. The outcome differed between trials, but diagnosis and disease status pre-transplant significantly influenced outcome. In summary, this approach is feasible and provides access to the curative potential of allogeneic stem cell transplantation for patients with higher age or comorbidities. Since the majority of the reports included heterogeneous patient populations, mostly with a short follow-up, more and specifically randomized studies are needed to define the role of RIC before allogeneic hematopoietic cell transplantation. PMID- 12910374 TI - Recombinant human erythropoietin, epoetin beta, in patients with relapsed lymphoma treated with aggressive sequential salvage chemotherapy--results of a randomized trial. AB - The aim of the study was to investigate the effects of erythropoietin (epoetin beta) on red blood cell (RBC) transfusions, hemoglobin (Hb) levels, and quality of life (QOL) in patients with relapsed lymphoma treated with an aggressive sequential salvage chemotherapy (SSCT) regimen. Sixty patients with early or late relapsed Hodgkin's disease ( n=39) or first relapse of aggressive non-Hodgkin's lymphoma ( n=21) were randomized to receive epoetin beta 10,000 IE subcutaneously three times a week or no epoetin during salvage chemotherapy. Patients in both study arms received two cycles of DHAP (dexamethasone, high-dose cytarabine, cisplatin); patients in partial remission (PR) or complete remission (CR) then received cyclophosphamide, followed by peripheral blood stem cell (PBSC) harvest, methotrexate plus vincristine, and etoposide. The final myeloablative course was BEAM (carmustine, etoposide, cytarabine, and melphalan) followed by autologous stem cell support. The primary endpoint of the study was the number of RBC units needed during SSCT. In addition, Hb levels and QOL were measured. The mean number of RBC units given in the epoetin beta arm was 4.5 compared to 8.3 in the control arm ( P=0.0134). The mean Hb levels during therapy were 10.4 g/dl in the epoetin beta arm and 9.7 g/dl in the control ( P=0.018). From baseline until BEAM therapy QOL (EORTC QLQ C30) and fatigue (MFI) assessment showed little QOL worsening or stable levels in both arms with a steeper increase of fatigue levels in the control group. Patients with relapsed lymphoma undergoing aggressive chemotherapy and stem cell support benefited from epoetin beta therapy, with a decrease of RBC transfusion requirements and lower rise of fatigue levels. PMID- 12910375 TI - Salvage chemotherapy according to the ASHAP protocol: a single-center study of 24 patients with relapsed or refractory aggressive non-Hodgkin's lymphomas. AB - Numerous salvage protocols for relapsed or refractory aggressive non-Hodgkin's lymphomas have been described. The purpose of this retrospective study was to evaluate the efficacy and the toxicity of the ASHAP protocol, which combines a continuous infusion of doxorubicin and cisplatin with high-dose cytarabine and methylprednisolone. Twenty-four patients with relapsed or refractory aggressive non-Hodgkin's lymphomas were treated with a median of 3 cycles (range: 1-5) of ASHAP. Twelve patients achieved a complete and four a partial remission for an overall response rate of 67%. The 3-year overall and progression-free survival rates were 60% and 40%, respectively. Ten of the responding patients were consolidated by high-dose chemotherapy. After a median follow-up of 15.5 months, four patients are in continuous complete remission, while six patients suffered relapses (two fatal). For reasons of low risk profile [international prognostic index (IPI) score of 0, n=2] or age >60 years ( n=4), consolidation was limited to involved-field radiotherapy in six patients. All of these patients are alive after a median follow-up of 37 months, with two relapses. Factors predicting a poor response to salvage therapy were primary refractory disease, elevated lactate dehydrogenase activity, and an IPI score of >/=2. The principal toxicity was myelosuppression with grade III or IV neutropenia or thrombocytopenia occurring in 88% or 75%, respectively, of the patients. Nonhematological toxicity was generally mild. There were no treatment-related deaths. The ASHAP regimen is a highly active and well-tolerated salvage protocol for patients with relapsed aggressive non-Hodgkin's lymphomas which compares favorably with other established protocols. PMID- 12910376 TI - Telomere length shortening in non-Hodgkin's lymphoma patients undergoing chemotherapy. AB - We investigated telomere length changes in patients with non-Hodgkin's lymphoma (NHL) receiving conventional-dose chemotherapy. Using Southern blot analysis, telomere length was measured in peripheral blood mononuclear cells from five NHL patients at diagnosis, 15 NHL patients after chemotherapy, and 39 healthy controls. Compared with age-matched putative normal controls, telomeres were significantly shorter in NHL patients at diagnosis. Mean telomere length was shorter after chemotherapy than before chemotherapy and was shorter after chemotherapy than in age-matched putative healthy controls. There was no correlation between the extent of telomere shortening and time elapsed after chemotherapy. These findings suggest that in NHL patients hematopoietic stem cells lose telomere length during the recovery period from bone marrow suppression after conventional-dose chemotherapy. PMID- 12910377 TI - Terminal differentiation in vitro of patient-derived post-TMD megakaryoblastic AML cells. AB - Differentiation induction is a therapeutic principle in acute promyelocytic leukemia (AML) using all- trans retinoic acid. In cell lines with properties of AML M6/M7 (K562 and CMK), differentiation towards megakaryopoietic and erythropoietic phenotypes can be induced in vitro. Transitory myeloproliferative disorder (TMD) is a self-limited disorder of newborn infants with Down syndrome, phenotypically resembling acute myeloid leukemia of megakaryoblastic lineage. Despite spontaneous disappearance of blasts from blood and bone marrow, in about 10% of the patients, overt acute megakaryoblastic leukemia (AML M7) develops up to 4 years later. Recently, mutations of the GATA1 transcription factor have been identified in the megakaryoblastic leukemia of Down syndrome. Here, we studied cells from a patient suffering from megakaryoblastic AML at the age of 2.5 years after spontaneous remission of neonatal TMD. In vitro, terminal differentiation towards a megakaryocyte-like phenotype could be induced by phorbol myristate acetate (PMA), with typical morphological features, upregulation of platelet specific and downregulation of erythroid antigens, going along with downregulation of c-myc. Whether spontaneous resolution of TMD is a process due to terminal differentiation is still open; however, here we give evidence that in vitro differentiation can be induced even in blasts deriving from an overt AML French-American-British (FAB) M7 after TMD. PMID- 12910378 TI - Subaortic left brachiocephalic vein: computed tomography and magnetic resonance angiography findings. AB - A case of anomalous (subaortic) position of the left brachiocephalic vein was incidentally detected on computed tomography images. Magnetic resonance angiography was performed to demonstrate the relationship of this vessel with other vascular structures. The anomalous vein was formed by the union of the left internal jugular and left subclavian veins. This vein passed downward along the left lateral side of the aortic arch, entered the aorticopulmonary window, descended in the mediastinum between the ascending aorta and the trachea and joined with the right brachiocephalic vein to form the superior vena cava. No cardiac anomalies accompanied the subaortic left brachiocephalic vein in the present case. We present the computed tomography and magnetic resonance angiography findings of this rare anomalous vein. PMID- 12910379 TI - Right internal iliac vein joining the left common iliac vein: case report demonstrated by CT angiography. AB - An unusual variation of the iliac veins was detected by computed tomography (CT) angiography in a 35-year-old man. In coronal CT reconstructions, it was shown that the right internal iliac vein of this patient crossed to the left side and drained to the left common iliac vein. This variation is important in retroperitoneal, laparoscopic and orthopedic surgery. We present the CT findings and discuss the embryological origin of this unusual congenital anomaly. PMID- 12910380 TI - Ectopic stapes: a case report with embryologic correlations. AB - A case report of unilateral congenital stapes misplacement revealed by computed tomography is presented. In addition to this malformation, the malleus was synostosed to the middle ear roof. This kind of stapes ectopia has not been described previously. We have analyzed the malformative pattern in the light of normal and teratologic development of the stapes. In a teratologic model in which retinoic acid is administered to pregnant mice, we have observed an ectopic stapes primordium independent of the otic capsule. We discuss the possible pathogenesis of this abnormality in terms of the genetic events of middle ear development, which can be perturbed by retinoic acid administration. PMID- 12910381 TI - The inferior laryngeal nerve: surgical and anatomic considerations. Report of 251 thyroidectomies. AB - This surgical anatomic study aimed to determine (1) the anatomic relation of the laryngeal inferior nerve with the inferior thyroid artery, (2) the existence of extralaryngeal branches of division of the nerve and (3) the size of the nerve seen macroscopically. Two hundred and fifty-one patients underwent thyroid surgery during a period of 30 months. There were 50 males and 201 females. The male population underwent 28 total thyroidectomies, 13 left lobectomies and 9 right lobectomies. The female population underwent 124 total thyroidectomies, 33 left lobectomies and 44 right lobectomies. On the right side: the nerve was found superficial to the artery in 70.24% of females and 51.35% of males, the nerve was divided in 23.81% of females and 21.62% of males and seemed unusually thin in 14.29% of females and 5.41% of males. On the left side: the nerve was found superficial to the artery in 87.26% of females and 95.12% of males, the nerve was divided in 15.29% of females and 14.63% of males and seemed unusually thin in 10.83% of females and 2.44% of males. In conclusion, the inferior laryngeal nerve is characterized by its important anatomic variations, especially on the right side. These variations might be different even between males and females. Knowledge of these variations is very important in order to best identify and preserve the inferior laryngeal nerve during thyroid surgery. PMID- 12910382 TI - Classical and nerve-sparing radical hysterectomy: an evaluation of the risk of injury to the autonomous pelvic nerves. AB - Radical hysterectomy represents the treatment of choice for FIGO stage IA2-IIA cervical cancer. It is associated with several serious complications such as urinary and anorectal dysfunction due to surgical trauma to the autonomous nervous system. In order to determine those surgical steps involving the risk of nerve injury during both classical and nerve-sparing radical hysterectomy, we investigated the relationships between pelvic fascial, vascular and nervous structures in a large series of embalmed and fresh female cadavers. We showed that the extent of potential denervation after classical radical hysterectomy is directly correlated with the radicality of the operation. The surgical steps that carry a high risk of nerve injury are the resection of the uterosacral and vesicouterine ligaments and of the paracervix. A nerve-sparing approach to radical hysterectomy for cervical cancer is feasible if specific resection limits, such as the deep uterine vein, are carefully identified and respected. However, a nerve-sparing surgical effort should be balanced with the oncological priorities of removal of disease and all its potential routes of local spread. PMID- 12910384 TI - Violent coalitionary attacks and intraspecific killing in wild white-faced capuchin monkeys ( Cebus capucinus). AB - During 12 years of observation, we have observed three confirmed and two inferred lethal coalitionary attacks on adult male white-faced capuchins ( Cebus capucinus) by members of two habituated social groups at Lomas Barbudal Biological Reserve, Costa Rica. In one case, an alpha male was badly wounded and evicted from his group, and when later found by his former groupmates he was attacked by several of them and died less than 24 h later. In two other cases, lone extra-group males were mobbed by adult and immature males of a bisexual group. One victim's abdomen was torn open and he died less than 24 h later. A second victim was quite badly bitten but may have escaped. The fourth and fifth cases resulted from intergroup encounters. One victim lost the use of both arms but may have survived, whereas the other died of unknown causes within an hour of the attack. The observed death rate from coalitionary aggression at our site is approximately the same as that reported for eastern chimpanzees. Because at least three of the five observed incidents involved large coalitions attacking lone victims, they support the general hypothesis that imbalances of power contribute to intraspecific killing in primates. However, the occurrence of lethal coalitional attacks in a species lacking fission-fusion social organization poses a challenge to the more specific version of the imbalance-of-power hypothesis proposed by Manson and Wrangham in 1991 to explain chimpanzee and human intergroup aggression. PMID- 12910385 TI - Stress proteins of Clostridium perfringens type A immunoreact with antiserum from rabbits infected with gas gangrene. AB - Various stressors were used to induce stress proteins in Clostridium perfringens. Cultures of C. perfringens FD-1041 were subjected to cold shock (28 degrees C for 1 h), acid shock (pH 4.5 for 30 min), or heat shock (50 degrees C for 30 min). Cells were lysed and protein samples were analyzed by immunoblotting with antiserum derived from rabbits suffering from gas gangrene. Eight cold shock proteins (approximate Mr 101, 82, 70, 37, 22, 12, 10 and 6 kDa) and also eight heat shock proteins (approximate Mr 101, 82, 70, 27, 22, 16, 12 and 10 kDa) were immunoreactive with the serum. No immunoreactive proteins were detected in samples subjected to acid shock proteins and purified DnaK protein was also non immunoreactive with the serum. These immunogenic stress proteins may be important in regulating diseases caused by C. perfringens. Such proteins could be involved in cell survival mechanisms, serve as targets during infection, or play a role in recognition of the bacteria by the host. PMID- 12910386 TI - Cultivation and properties of Echinamoeba thermarum n. sp., an extremely thermophilic amoeba thriving in hot springs. AB - Here we describe a new, extremely thermophilic amoeba growing between 33 degrees C and 57 degrees C ( Topt.=50 degrees C). Isolates had been obtained from hot springs at Agnano Terme (Italy), Yellowstone National Park (USA), Kamchatka (Russia), and the Arenal Volcano (Costa Rica). They could be cultured monoxenically on a thermophilic alpha-proteobacterium. The morphology of the amoeba was studied using a microscope situated under a heatable polyacrylate hood. At 50 degrees C, the cells appeared flat with an irregular triangular or elongate shape, sometimes exhibiting fine spine-like subpseudopodia. On average, they were 22 microm long and 11 microm wide and had one nucleus with a central nucleolus. Based on morphology and on SSU rRNA comparisons, the amoeba belonged to the genus Echinamoeba, where it represents a new species. Referring to its extremely thermophilic lifestyle and its hydrothermal habitat, we name it E. thermarum. PMID- 12910383 TI - Scintimammography with 99mTc-MIBI and magnetic resonance imaging in the evaluation of breast cancer. AB - This study was performed to evaluate the sensitivity and specificity of technetium-99m methoxyisobutylisonitrile (99mTc-MIBI) scintimammography (SMM) and contrast-enhanced magnetic resonance imaging (MRI) in patients with breast masses, using the histological findings as the gold standard. Forty-five consecutive patients with a breast lesion, detected by self-examination, physical examination or screening mammography, underwent SMM and MRI. In 38 cases (84.5%), the histopathology was malignant; the breast cancers ranged from 3 to 100 mm in diameter (mean 22 mm). In the overall patient group, MRI showed a slightly higher sensitivity than SMM (92% vs 84%), but SMM showed a better specificity: 71% vs 42%. The accuracy was 82% and 84% for SMM and MRI respectively. To evaluate the influence of lesion size on the results, patients with lesions < or =20 mm and < or =15 mm were examined. In patients with lesions < or =20 mm, the sensitivity of SMM and MRI decreased to 64% and 82% respectively, while SMM again displayed considerably better specificity: 83% vs 50% for MRI. The accuracy of SMM and MRI was 64% and 82% respectively. In patients with lesions < or =15 mm, SMM again showed better specificity (75% vs 50%), while MRI displayed better sensitivity and accuracy (sensitivity, 81% vs 62%; accuracy, 75% vs 65%). In this study the specificity of SMM in patients with breast lesions was thus superior to that of MRI. The combination of SMM and MRI may be used in those patients with equivocal findings at mammography and ultrasound to reduce the number of unnecessary surgical biopsies. PMID- 12910387 TI - High 16S rDNA bacterial diversity in glacial meltwater lake sediment, Bratina Island, Antarctica. AB - The microbial diversity in maritime meltwater pond sediments from Bratina Island, Ross Sea, Antarctica was investigated by 16S rDNA-dependent molecular phylogeny. Investigations of the vertical distribution, phylogenetic composition, and spatial variability of Bacteria and Archaea in the sediment were carried out. Results revealed the presence of a highly diverse bacterial population and a significantly depth-related composition. Assessment of 173 partial 16S rDNA clones analyzed by amplified rDNA restriction analysis (ARDRA) using tetrameric restriction enzymes (HinP1I 5'G/CGC3'and Msp I. 5'C/CGG3', BioLabs) revealed 153 different bacterial OTUs (operational taxonomic units). However, only seven archaeal OTUs were detected, indicating low archaeal diversity. Based on ARDRA results, 30 bacterial clones were selected for sequencing and the sequenced clones fell into seven major lineages of the domain Bacteria; the alpha, gamma, and delta subdivisions of Proteobacteria, the Cytophaga-Flavobacterium Bacteroides, the Spirochaetaceae, and the Actinobacteria. All of the archaeal clones sequenced belonged to the group Crenarchaeota and phylogenetic analysis revealed close relationships with members of the deep-branching Group 1 Marine Archaea. PMID- 12910388 TI - A novel lineage of sulfate-reducing microorganisms: Thermodesulfobiaceae fam. nov., Thermodesulfobium narugense, gen. nov., sp. nov., a new thermophilic isolate from a hot spring. AB - A novel type of a sulfate-reducing microorganism, represented by strain Na82T, was isolated from a hot spring in Narugo, Japan. The isolate was a moderate thermophilic autotroph that was able to grow on H2/CO2 by sulfate respiration. The isolate could grow with nitrate in place of sulfate, and possessed menaquinone-7 and menaquinone-7(H2) as respiratory quinones. Phylogenetic analysis of the 16S rRNA gene sequence indicated that strain Na82T was a member of the domain Bacteria and distant from any known bacteria, as well as from other sulfate-reducing bacteria (sequence similarities less than 80%). The phylogenetic analysis of the dsrAB gene (alpha and beta subunits of dissimilatory sulfite reductase) sequence also suggested that strain Na82T was not closely related to other sulfate reducers. On the basis of the phenotypic and phylogenetic data, a new taxon is established for the isolate. We proposed the name Thermodesulfobium narugense gen. nov., sp. nov. with strain Na82T (=DSM 14796T=JCM 11510T) as the type strain. Furthermore, a new family, Thermodesulfobiaceae fam. nov., is proposed for the genus. PMID- 12910389 TI - Piezophysiology of genome wide gene expression levels in the yeast Saccharomyces cerevisiae. AB - Hydrostatic pressure is one of the physical factors affecting cellular physiology. Hydrostatic pressure of a few hundred MPa decreases the viability of yeast cells, and pressure of a few tens MPa decreases the growth rate. To understand the effect of hydrostatic pressure, we employed yeast DNA microarrays and analyzed genome-wide gene-expression levels after the pressure treatment with 180 MPa (immediate) at 4 degrees C and recovery incubation for 1 h and 40 MPa (16 h) at 4 degrees C and recovery incubation for 1 h. The transcription of genes involved in energy metabolism, cell defense, and protein metabolism was significantly induced by the pressure treatment. Genome-wide expression profiles suggested that high pressure caused damage to cellular organelles, since the induced gene products were localized in the membrane structure and/or cellular organelles. Hierarchical clustering analysis suggested that the damage caused by the pressure was similar to that caused by detergents, oils, and freezing/thawing. We also estimated the contribution of induced genes to barotolerance using some strains that have the deletion in the corresponding genes. PMID- 12910390 TI - Alpha-amylase activity from the halophilic archaeon Haloferax mediterranei. AB - The halophilic archaeon Haloferax mediterranei is able to grow in a minimal medium containing ammonium acetate as a carbon and nitrogen source. When this medium is enriched with starch, alpha-amylase activity is excreted to the medium in low concentration. Here we report methods to concentrate and purify the enzyme. The relative molecular mass of the enzyme, determined by gel filtration, is 50 +/- 4 kDa, and on SDS-PAGE analysis a single band appeared at 58 kDa. These results indicated that the halophilic alpha-amylase is a monomeric enzyme. The enzyme showed a salt requirement for both stability and activity, being stable from 2 to 4 M NaCl, with maximal activity at 3 M NaCl. The enzyme displayed maximal activity at pHs from 7 to 8, and its optimal temperature was in a range from 50 degrees C to 60 degrees C. The results also implicated several prototropic groups in the catalytic reaction. PMID- 12910391 TI - Mauran, an exopolysaccharide produced by the halophilic bacterium Halomonas maura, with a novel composition and interesting properties for biotechnology. AB - Mauran is an anionic, sulfated heteropolysaccharide with a high uronic-acid content, synthesized by strain S-30 of the halophilic bacterium Halomonas maura. Under optimum environmental and nutritional conditions, it is capable of producing up to 3.8 g of mauran per liter of medium. Aqueous solutions of mauran are highly viscous and display pseudoplastic, viscoelastic and thixotropic behavior. Its viscosity is stable over a wide pH range (3-11), after freezing thawing processes, and in the presence of sucrose, salts, surfactants and alpha hydroxyl acids. It has a high capacity for binding lead and other cations. Its molecular mass when collected from an MY medium supplemented with 2.5% w/v salt during the stationary growth phase is 4.7x10(6) Da. PMID- 12910392 TI - Cold-active serine alkaline protease from the psychrophilic bacterium Pseudomonas strain DY-A: enzyme purification and characterization. AB - An extracellular protease was purified from a deep-sea psychrophilic bacterium strain DY-A which was identified as a Pseudomonas species. The optimal growth and protease-producing temperatures of the strain were all 10 degrees C, and the protease was secreted only at temperatures under 20 degrees C. The enzyme was most active at 40 degrees C and at pH 10.0. It was inhibited by phenylmethyl sulfonylfluoride and diisopropyl fluorophosphate, indicating that it is a serine protease. Chelators such as EDTA, EGTA, 1,10-phenanthroline and 2,2'-bipyridyl produced a decrease of activity. The enzyme was sensitive to denaturing agents such as SDS, urea, and guanidine HCl and resistant to thiol-containing reducing agents such as dithiotreitol. The enzyme was active towards N-succinyl-Ala-Ala Pro-Phe- p-nitroanilide and N-succinyl-Ala-Ala-Pro-Leu- p-nitroanilide. The native molecular mass of the enzyme determined by native PAGE and SDS-PAGE was 25 kDa. PMID- 12910393 TI - Harmonisation of neurosurgical training in the European Countries--a personal view. PMID- 12910394 TI - Pressure-autoregulation, CO2 reactivity and asymmetry of haemodynamic parameters in patients with carotid artery stenotic disease. A clinical appraisal. AB - OBJECTIVES: Patients with carotid artery stenotic disease and poor cerebral haemodynamic reserve are in increased risk of stroke. Haemodynamic reserve can be estimated by measuring cerebrovascular reactivity induced by breathing CO2 and pressure-autoregulation by analyzing spontaneous slow fluctuation in arterial pressure and MCA blood flow velocity. We evaluated the relationship between clinical status, CO2 reactivity, pressure-autoregulation and other haemodynamic variables derived from Transcranial Doppler ultrasonography in patients with carotid artery disease. METHODS: 38 patients were investigated. Arterial pressure and blood flow velocity were monitored during CO2 reactivity tests. Arterial pressure-corrected and non-corrected indices of CO2 reactivity were calculated to compare an influence of rise in arterial pressure during the test. The pressure autoregulation index was calculated as correlation coefficient between slow waves in blood flow velocity and arterial pressure. RESULTS: The increase in CO2 produced a consistent rise in arterial pressure and blood flow velocity and weakened the pressure autoregulation. The value of pressure-corrected CO2 reactivity was lower (p<0.0001) than the non-corrected one, indicating that the rise in arterial pressure during the test tends to over-estimate CO2 reactivity. The pressure-corrected reactivity was correlated with pressure autoregulation (r= 0.46; p<0.005). Both CO2 reactivity and pressure-autoregulation index correlated with a degree of carotid artery stenosis. Side-to-side difference of TCD pulsatility index demonstrated a close relationship with the asymmetry of stenosis (r=-0.61; p<0.0002) and symptoms (r=-0.49; p<0.003). CONCLUSIONS: When calculating CO2 reactivity in patients with carotid artery disease, changes in arterial pressure should be considered. Both CO2 reactivity and pressure autoregulation describe the magnitude of haemodynamic deficit caused by stenosis, pulsatility index expresses the asymmetry of stenosis. PMID- 12910395 TI - Risk factors for the occurrence of chronic subdural haematomas after neurosurgical procedures. AB - Chronic subdural haematoma (CSDH) is a rare clinical complication of neurosurgical procedures. CSDH occurs sporadically after aneurysm clipping surgery and revascularisation surgery but the risk factors are not known. The present study reviewed 6613 consecutive neurosurgical procedures performed from January 1987 to July 2001, and identified 621 cases of CSDH. Fifty of these 621 cases had a past history of neurological disorders treated by neurosurgery. This study evaluated these 50 cases in order to elucidate the clinical and radiological characteristics of CSDH after neurosurgery and to investigate the etiology for identifying in the risk factors of CSDH as a postoperative complication. The incidence of CSDH after neurosurgery was 0.8% (50/6613). Twenty seven of the 50 patients with a past history of neurosurgery had undergone aneurysm clipping surgery. The incidence after clipping surgery was 2.4%. Twelve of these 27 cases also underwent ventriculoperitoneal shunting. Three patients had postoperative CSDH after arachnoid cyst opening and/or shunting. The incidence was highest at 7.5%. Three patients had postoperative CSDH after brain tumour surgery. The incidence was low at 0.4%. However, the ventricular cerebrospinal fluid (CSF) space was opened during tumour removal in 2 of these 3 patients. Communication of the subarachnoid space to the subdural space is considered to be one of the causative factors and excessive CSF shunting facilitates formation of CSDH after neurological surgery. Repair of arachnoid tearing during neurosurgery and avoidance of excessive CSF shunting may reduce the risk of CSDH after neurosurgery. PMID- 12910396 TI - A clinical comparison of non-traumatic acute subdural haematomas either related to coagulopathy or of arterial origin without coagulopathy. AB - BACKGROUND: Non-traumatic acute subdural haematomas enable study of the morbidity and mortality due to the haematoma without the effect of trauma. Whereas it is known that coagulation disorders worsen the outcome of spontaneous intracerebral haematomas, this has not been studied in non-traumatic acute subdural haematomas. METHODS: In a series of 13 non-traumatic acute subdural haematomas admitted to our department between January 1995 and March 2002, we had 9 coagulopathy associated haematomas and 3 haematomas corresponding to the syndrome of 'spontaneous acute subdural haematoma of arterial origin'. Both groups were compared. FINDINGS: Age and gender distribution were comparable. The bleeding source was a cortical artery in 2 of the 2 non-coagulopathy related haematomas operated on, but also in 2 of the 4 coagulopathy associated haematomas that underwent surgery. The average haematoma thickness was higher in the coagulopathy related haematomas. The mean Glasgow Coma Score on admission was 7.7 and the mortality rate was 55.6% in the coagulopathy related group. In the non coagulopathy related haematomas the mean Glasgow Coma Score was 12.0 and the mortality rate 33.3%. The latter mortality rate corresponds well to that of a historical group of 'spontaneous acute subdural haematomas of arterial origin' collected from the literature. INTERPRETATION: The outcome was worse in the non traumatic acute subdural haematomas that were associated with a coagulation deficiency. While in all non-traumatic acute subdural haematomas the interval to surgery should be minimized, early recognition and urgent correction of coagulation deficiencies is certainly indicated. PMID- 12910397 TI - Stereotactic biopsy for intracranial lesions: reliability and its impact on the planning of treatment. AB - BACKGROUND: The authors present a retrospective analysis of 308 computed tomography (CT)-guided stereotactic biopsies in 300 patients in order to evaluate the reliability and efficacy of the stereotactic biopsy for intracranial lesions. METHOD: All patients were suffering from undetermined intracranial lesions and treated at Seoul National University Hospital between January 1993 and December 1999. Age ranged from three to 79 years (mean 41); the male to female ratio was 180:120. All patients underwent CT-guided stereotactic biopsy for the histological verification and/or evacuation of the cyst using Riechert-Mundinger stereotactic system. FINDINGS: Histological diagnosis was made in 275 patients (diagnostic yield 91.7%). Diagnostic yield was better in group with frozen section examination during the stereotactic procedure than the group without it (p=0.01). Neoplastic lesions were more likely to be diagnosed in stereotactic biopsy than non-neoplastic lesions (p=0.02). Among 30 patients who underwent craniotomy after the stereotactic biopsy, the histological diagnoses after the craniotomy were identical to those of the stereotactic biopsy in 29 patients (diagnostic accuracy 96.7%). Two patients died within seven days after the stereotactic biopsy (mortality 0.6%). The postoperative new neurological deficit or aggravation of the neurological status was found in 19 patients, including transient cases of seven patients (permanent morbidity rate 3.9%). Histologically malignant gliomas and deeply-located lesions were the significant risk factors for the development of complications. In 148 cases, histological diagnosis of the stereotactic biopsy was different from the preoperative clinical diagnosis. Among these cases, the treatment plan was changed after stereotactic biopsy in 81 cases. CONCLUSIONS: Stereotactic biopsy for intracranial lesions is a reliable and relatively safe procedure. It is also a very efficacious method especially in patients who need histological confirmation for the treatment. PMID- 12910398 TI - Diagnosis and treatment of progressive space-occupying radiation necrosis following stereotactic radiosurgery for brain metastasis: value of proton magnetic resonance spectroscopy. AB - BACKGROUND: There have been some reports that radiation necrosis can be controlled conservatively. There are rare cases showing progressive space occupying radiation necrosis (PSORN). It is very difficult to control PSORN by conservative treatment. The purpose of this study was to evaluate the early diagnosis of those cases and the timing of surgery for patients with PSORN. METHOD: We have experienced some cases where quality of life was improved by the removal of PSORN after stereotactic radiosurgery (SRS) for brain metastases. Therefore, we evaluated retrospectively the diagnosis and treatment of six cases of symptomatic PSORN at approximately 6-12 months after SRS for metastatic brain tumours. FINDINGS: In all six cases, on Magnetic Resonance Imaging with Gd contrast material (Gd-MRI), PSORN was revealed as a ring-like enhanced mass with large perifocal oedema coupled with the appearance of neurological deficit. Proton Magnetic Resonance Spectroscopy ((1)H-MRS) enabled us to differentiate PSORN from recurrence of metastases in all six cases. Single Photon Emission Computed Tomography with thallium-201 chloride (201TlCl-SPECT) enabled us to do this in four cases of the six. In four cases of the six, lesionectomy of the ring like enhanced mass (PSORN) was performed, and in two of these cases the removal was performed within 4 weeks from the time when conservative treatment became ineffective, and the neurological deficit and perifocal oedema was improved as was the quality of life. However, in the other two patients who were left for more than 16 weeks, the deficit was gradually progressive. The two patients who did not receive lesionectomy were treated by conservative means with steroids and/or heparin and warfarin and they had progressive neurological symptoms. INTERPRETATION: Although, the number of patients is small in this study, and more data will be needed, it is recommended that lesionectomy is performed at an early stage, if possible, when conservative management has failed. PMID- 12910399 TI - Disc height loss after anterior cervical microdiscectomy with titanium intervertebral cage fusion. AB - BACKGROUND: The use of a graft in cervical inter-vertebral disc repair is still a controversial procedure. The aim of the treatment is to restore the physiological disc height and to achieve fusion. This study was performed to determine the rate of narrowing of the cervical intervertebral disc after using a titanium cervical intervertebral cage (BAK-C; Sulzer Spine Tech, Minneapolis, MN). METHODS: 43 patients were included in the study. Each had a cervical disc protrusion, resulting in radiculopathy. All underwent surgery involving an anterior one level cervical microdiscectomy and fusion with a cylindrical titanium cage. The mean follow-up time was 18+/-5 months. FINDINGS: The extent of the mean narrowing rate of disc space was 35.6+/-9%. The fusion rate was 98% and the mean time to fusion was 6 months. INTERPRETATION: The use of a cervical intervertebral cage in anterior cervical microdiscectomy does not prevent the loss of the height of the cervical disc space after the operation. PMID- 12910400 TI - Changes in vascular responses of the basilar artery to acetylcholine and endothelin-1 in an experimental rabbit vasospasm model. AB - BACKGROUND: This study aimed to clarify the characteristics of the basilar arteries of rabbits two and seven days after subarachnoid haemorrhage (SAH) and to investigate the influence of hypoxia on vascular responses to acetylcholine and endothelin-1. METHODS: Thirty four New Zealand white rabbits were divided into 3 groups: 1) control (n=6), 2) 2 days post SAH (n=15), and 3) 7 days post SAH (n=13). Rabbits in the groups 2 and 3 underwent vertebral angiography before sacrifice. Harvested vascular rings were suspended in a small vessel myograph system for isometric tension recording in response to vasodilatory and vasoconstrictory stimulation in oxygenated (95% O2/5% CO2) and hypoxic (95% N2/5% CO2) conditions. To investigate the response to a vasodilator, acetylcholine (ACh) (10(-7) to 3 x 10(-4) M) was applied to a basilar artery pre-contracted with histamine (10(-6) approximately 10(-5) M). To investigate the response to a vasoconstrictor, the effect of endothelin-1 (ET-1) (10(-11) to 3 x 10(-8) M) was observed in quiescent rings. FINDINGS: Seven of 15 rabbits that underwent angiography two days after operation showed vasospasm to 64.3+/-11.2% of the original diameter; likewise seven of the 13 rabbits studied seven days after operation showed vasospasm to 64.9+/-10.9% of the original diameter. Hypoxic conditions significantly reduced the relaxation response to ACh but did not influence basilar artery contraction to ET-1. Responses to ACh and ET-1 did not differ significantly in vessels from animals with or without vasospasm at 2 days. In contrast, at 7 days vascular responses to ACh and ET-1 were significantly less in vessels from animals with vasospasm than those without (p=0.029, 0.032), and also less than in vessels from animals with vasospasm at day 2 (p=0.002, 0.004). There was not a significant difference in the vascular responses of basilar arteries from animals without vasospasm. INTERPRETATION: The markedly lower vascular responses of basilar arteries of rabbits to ACh and ET-1 found seven days after SAH may relate to delayed vasospasm in patients with SAH. PMID- 12910401 TI - Intra-arterial simultaneous administration of anandamide attenuates endothelin-1 induced vasospasm in rabbit basilar arteries. AB - BACKGROUND: Anandamide induces not only endothelium-dependent vasodilatation through cannabinoid receptors but also some endothelium- independent vasodilator effect by calcitonin gene-related peptide release through vanilloid receptors. Endothelin-1, a powerful vasoconstrictive peptide derived from endothelial cells, has been shown to be converted to its active form after cleaving by a vascular matrix metalloproteinase which is also involved in inactivation of calcitonin gene-related peptide. The purpose of this study was to investigate whether anandamide inhibits the acute vascular and morphological effects of Endothelin-1 applied intra-arterially on rabbit basilar arteries. METHOD: Fifteen albino rabbits were anaesthetised and underwent placement of a vertebral artery catheter for angiography of the basilar artery. Animals were divided, arbitrarily, into animals in which there was either intra-arterial injection of saline (Group I, n=5), Endothelin-1 (Group II, n=5) and Endothelin-1 and anandamide (Group III, n=5). The diameter of the basilar artery between the pre and post injection angiograms was measured in each of the three groups and transmission electron microscopic investigations on basilar arteries were performed. FINDINGS: Angiographic studies showed that simultaneous administration of anandamide significantly attenuated Endothelin-1 induced vasoconstriction. Furthermore, it was demonstrated that anandamide reversed the morphological changes induced by Endothelin-1 on the vessel wall. INTERPRETATION: These results indicated that anandamide overcomes the angiographic and morphological effects of intrarterially administered ET-1 induced vasospasm in rabbit basilar arteries probably by induction of CGRP related vasodilatation through vanilloid receptors and prevents the acute ET-1 induced ultrastructural vessel wall damage. PMID- 12910402 TI - "Basal durotomy" to prevent massive intra-operative traumatic brain swelling. AB - OBJECTIVE: Massive intraoperative swelling (IOS) is a potential complication of intracranial surgery for traumatic hematomas. We present a novel design of dural opening to minimize the risks of IOS. METHODS: Over the last eight years, we have used and evaluated a "reversed U-shaped" durotomy incision, which leaves the dura intact in the parasagittal region. Accordingly, the dura should be opened initially over the fronto-basal eminence of the frontal lobe anterior to the middle meningeal artery and sylvian region. The incision is then carried across the middle meningeal into the temporal region. Additional slit incisions may be made, as needed, around the circumference of the craniotomy in order to obtain sufficient access to remove a subdural hematoma. RESULTS: We have employed this technique in 220 trauma craniotomies without any case of uncontrolled IOS. In all patients a complete dural closure was obtained. CONCLUSION: When IOS is suspected, basal durotomy proved to be an excellent technique to prevent it, without compromising the total evacuation of the traumatic intracranial hematoma. PMID- 12910403 TI - Meningio-angiomatosis in a patient with focal epilepsy: value of PET in diagnoses and preoperative planning of surgery. AB - A 17-year-old boy with epileptic seizures due to meningio-angiomatosis without neurofibromatosis type 2 is presented. Low grade astrocytoma in the left temporal lobe was resected when he was 11 years old. A recurrence was suspected on following-up MRI and a positive PET scan with 11C-methionine PET 6 years later around the resected area. The language area was mapped using H2(15)O PET activation technique. The lesion was completely resected while preserving the verbal area assisted by three-dimensional imaging protocol of MR-registered PET. The patient was well and seizure-free for 8 years thereafter without antiepileptic drugs. Histologically, there was an increase of dilated arterioles and meningothelial cell-like spindle cells around them, which are characteristic pathological features of meningio-angiomatosis. It is most likely that angiomatous tissue developed perivascular meningiomatous components and formed the meningio-angiomatosis in our presented case. In addition, we presented our protocol of functional neuro-imaging which was useful in terms of the functional neurosurgery. PMID- 12910404 TI - Mesenchymal chondrosarcoma of the sellar region. AB - BACKGROUND: It is known that, although rare, mesenchymal chondrosarcoma can originate intracranially. However, no such malignant tumour has been described in the sellar region. CLINICAL PRESENTATION: We report a case of mesenchymal chondrosarcoma in a 21-year-old man who presented with double vision, right blepharoptosis and facial pain. Upon initial admission, no endocrinological abnormalities were found, and computed tomography and magnetic resonance imaging revealed a mass with calcification in the sella and right cavernous sinus. INTERVENTION: For this malignant tumour, three surgical resections, two sessions of gamma-knife radiosurgery, one session of fractional irradiation, and one cycle of chemotherapy were performed, resulting in only brief arrest of the tumour growth. Pathologically, the tumour consisted of undifferentiated small cells of high cellularity, and islands of hyaline cartilage. The undifferentiated small cells showed immunoreactivity for vimentin and ultrastructural features suggesting a mesenchymal origin. Lacunar cells in the islands were immunopositive for S-100 protein and vimentin. CONCLUSION: Although malignant tumours in the sellar region are rare, they should be considered in the differential diagnosis of various sellar tumours typified by non-functioning pituitary adenoma, and mesenchymal chondrosarcoma is one possible candidate. PMID- 12910405 TI - Intradural spinal seeding of a clival chordoma. AB - BACKGROUND: Following recent progress in radiological and surgical techniques for treating skull base tumours, more attention should be paid to the relatively rare phenomenon of intradural spinal seeding in cases of clival chordoma. CASE PRESENTATION: A 53-year-old man harboured intradural spinal seeding from a clival chordoma 11 years after his initial surgical treatment. He had undergone skull base surgery five times with extra- and intradural approaches, and adjuvant radiotherapy. His complaints of lumbago, paraparesis, and sensory disturbance of the lower left extremity prompted us to carry out imaging studies of the spinal cord, which revealed multiple intradural, extramedullary mass lesions at the thoracic and lumbosacral spinal level. The tumours were histopathologically diagnosed as spinal seeding of the clival chordoma with high proliferative potential (Ki-67 labelling index of 8.4%). INTERPRETATION: As long-term local control of skull base chordoma appears to be improving by the introduction of skull base surgical techniques and adjuvant radiotherapy, spinal intradural seeding might emerge as a potential complication. Close observation and timely imaging studies for spinal seeding of skull base chordoma would be required in long-term postoperative follow-ups, particularly for those patients with highly proliferative tumours. PMID- 12910406 TI - Orbital cystic neurinoma. AB - A 64-year-old woman presented with a 6-month history of exophthalmus of her right eye. She had no neurological deficit except for the exophthalmus and numbness in the right side of her forehead (frontal nerve territory). Neuro-imaging demonstrated a cystic mass in the right orbit. The tumour was totally removed by microsurgical transcranial surgery. The histological diagnosis was neurinoma. The origin of the tumour was considered to be the frontal nerve. Only two other cases of orbital cystic neurinoma have been reported. This rare clinical entity should be included in the differential diagnosis of a cystic mass in the orbit. PMID- 12910407 TI - Long-term oral Candida colonization, mucositis and salivary function after head and neck radiotherapy. AB - The aim of this study was to follow the long-term effects of radiation therapy of head and neck malignancies on oral yeast colonization, mucositis and salivary function. Included in this prospective study were 32 patients with intended radiation therapy of a malignancy of the head and neck. In all patients the salivary glands lay within the radiation field and the patients had at least five teeth. The first examination was performed after oral hygiene instruction and removal of questionable teeth before the start of radiotherapy. The following examinations were conducted after 3, 6, 9 and 12 months. Together with the quantitative determination of Candida colonization, three "mucositis" variables were assessed: (1) examiner-rated mucositis score (LENT/SOMA), (2) patient-rated mucositis symptoms, and (3) scintigraphic salivary excretion fraction. The maximum Candida colonization was found 6 months after radiation therapy and this declined to above normal values after 12 months. Salivary flow was at a minimum 6 months after radiation therapy and had slightly recovered by 12 months. Examiner rated mucositis and patient-rated xerostomia showed no significant recovery after 6 or 12 months. The results of this study show slight recovery of the oral ecological system. Although the causal role of a single parameter is not clear, persistently elevated Candida colonization should be taken into account therapeutically. PMID- 12910408 TI - Colony structure in a plant-ant: behavioural, chemical and genetic study of polydomy in Cataulacus mckeyi (Myrmicinae). AB - Social organisation of colonies of obligate plant-ants can affect their interaction with myrmecophyte hosts and with other ants competing for the resources they offer. An important parameter of social organisation is whether nest sites of a colony include one or several host individuals. We determined colony boundaries in a plant-ant associated with the rainforest understorey tree Leonardoxa africana subsp. africana, found in coastal forests of Cameroon (Central Africa). This myrmecophyte is strictly associated with two ants, Petalomyrmex phylax and Cataulacus mckeyi. Plants provide food and nesting sites for P. phylax, which protects young leaves against insect herbivores. This mutualism is often parasitised by C. mckeyi, which uses but does not protect the host. The presence of C. mckeyi on a tree excludes the mutualistic ant. Because Petalomyrmex-occupied trees are better protected, their growth and survival are superior to those of Cataulacus-occupied trees, giving P. phylax an advantage in occupation of nest sites. C. mckeyi often colonises trees that have lost their initial associate P. phylax, as a result of injury to the tree caused by disturbance. Polydomy may allow C. mckeyi to occupy small clumps of trees, without the necessity of claustral colony foundation in each tree. Investigating both the proximate (behavioural repertoire, colony odour) and the ultimate factors (genetic structure) that may influence colony closure, we precisely defined colony boundaries. We show that colonies of C. mckeyi are monogynous and facultatively polydomous, i.e. a colony occupies one to several Leonardoxa trees. Workers do not produce males. Thus, the hypothesis that polydomy allows workers in queenless nests to evade queen control for their reproduction is not supported in this instance. This particular colony structure may confer on C. mckeyi an advantage in short-distance dispersal, and this could help explain its persistence within the dynamic Leonardoxa system. PMID- 12910409 TI - Use of forest corridors by boreal Xestia moths. AB - We studied movements of the noctuid moths Xestia speciosa and X. fennica in a fragmented forest landscape including old-growth spruce forest corridors in eastern Finland. We individually marked and released 1,796 adults of the habitat generalist X. speciosa and 683 individuals of X. fennica, which prefers old growth spruce forests. Moths were recaptured with 60 sugar-bait traps that were placed within habitat corridors, in the non-habitat matrix and in surrounding old growth forest patches. Both species favoured the corridors during their movements and tended to avoid entering the matrix. We analysed recapture rates and movements of the recaptured moths. Results suggest that X. speciosa moves longer distances than X. fennica. Though corridors clearly canalized movements by both species, the impact of old-growth forest corridors at the larger landscape scale was less clear in X. fennica. Movement behaviour of the two sexes differed more than that of the two species. PMID- 12910410 TI - Effect of immersion in seawater on egg survival in the lizard Anolis sagrei. AB - Studies on the lizard, Anolis sagrei, revealed that after Hurricane Floyd devastated the Bahamas in 1999, some populations consisted only of hatchlings. Because the storm surge of the hurricane completely inundated these islands, apparently for up to 6 h, survival of anole eggs in salt water for such periods is implied. To test this hypothesis directly, we placed A. sagrei eggs in saltwater for 3 or 6 h with unimmersed eggs serving as the control. Hatching success and incubation time did not differ among the three treatments. These findings help explain the persistence of anole populations on small islands vulnerable to hurricanes. PMID- 12910411 TI - The Arabidopsis thaliana rlp mutations revert the ectopic leaf blade formation conferred by activation tagging of the LEP gene. AB - Activation tagging of the gene LEAFY PETIOLE (LEP) with a T-DNA construct induces ectopic leaf blade formation in Arabidopsis, which results in a leafy petiole phenotype. In addition, the number of rosette leaves produced prior to the onset of bolting is reduced, and the rate of leaf initiation is retarded by the activation tagged LEP gene. The ectopic leaf blade results from an invasion of the petiole region by the wild-type leaf blade. In order to isolate mutants that are specifically disturbed in the outgrowth of the leaf blade, second site mutagenesis was performed using ethane methanesulphonate (EMS) on a transgenic line that harbours the activation-tagged LEP gene and exhibits the leafy petiole phenotype. A collection of revertant for leafy petiole (rlp lines was isolated that form petiolated rosette leaves in the presence of the activated LEP gene, and could be classified into three groups. The class III rlp lines also display altered leaf development in a wild-type (non-transgenic) background, and are probably mutated in genes that affect shoot or leaf development. The rlp lines of classes I and II, which represent the majority of revertants, do not affect leaf blade outgrowth in a wild-type (non-transgenic) background. This indicates that LEP regulates a subset of the genes involved in the process of leaf blade outgrowth, and that genetic and/or functional redundancy in this process compensates for the loss of RLP function during the formation of the wild-type leaf blade. More detailed genetic and morphological analyses were performed on a selection of the rlp lines. Of these, the dominant rlp lines display complete reversion of (1) the leafy petiole phenotype, (2) the reduction in the number of rosette leaves and (3) the slower leaf initiation rate caused by the activation tagged LEP gene. Therefore, these lines are potentially mutated in genes for interacting partners of LEP or in downstream regulatory genes. In contrast, the recessive rlp lines exhibit a specific reversion of the leafy petiole phenotype. Thus, these lines are most probably mutated in genes specific for the outgrowth of the leaf blade. Further functional analysis of the rlp mutations will contribute to the dissection of the complex pathways underlying leaf blade outgrowth. PMID- 12910412 TI - Ultrastructure of spermiogenesis and the spermatozoon in the genus Joyeuxiella Fuhrmann, 1935 (Cestoda, Cyclophyllidea, Dipylidiidae): comparative analysis of J. echinorhynchoides (Sonsino, 1889) and J. pasqualei (Diamare, 1893). AB - This paper describes the ultrastructure of spermiogenesis and the mature spermatozoon of two Dipylidiidae cestodes, Joyeuxiella echinorhynchoides and J. pasqualei. In both species, spermiogenesis follows the type III described by Ba and Marchand for the cyclophyllideans. Nevertheless, it is interesting to note the presence of striated roots associated with the centrioles. The spermatozoon presents the same ultrastructural features in J. echinorhynchoides and J. pasqualei. The apical cone in the anterior extremity of the sperm is more than 2.0 microm long in both J. echinorhynchoides and J. pasqualei. Both species present a single crest-like body, 150 nm thick in J. echinorhynchoides and 75 nm thick in J. pasqualei. The cortical microtubules are spiralled at an angle of 40 45 degrees to the spermatozoon axis in both Joyeuxiella species. A periaxonemal sheath and glycogen granules are also described in the mature sperm. We also describe, for the first time, the disposition of glycogen granules in two opposed and spiralled cords in cestodes and the formation of the periaxonemal sheath in the final stage of spermiogenesis. PMID- 12910413 TI - Microarray analysis of changes in gene expression in a murine model of chronic chagasic cardiomyopathy. AB - Chagas' disease, caused by infection with Trypanosoma cruzi, is a major cause of cardiomyopathy in endemic regions. Infection leads to cardiac remodeling associated with congestive heart failure and dilated cardiomyopathy. In order to study the changes in the gene expression profile due to infection, C57BL/6 x 129sv male mice were infected with 1 x 10(3) trypomastigotes of the Brazil strain of T. cruzi. Histopathological examination of the myocardium revealed chronic inflammation, vasculitis and fibrosis 100 days post-infection. Cardiac magnetic resonance imaging revealed a significantly dilated heart compared with uninfected mice. The relative abundance or depletion of myocardial mRNAs was evaluated using high-density microarrays consisting of 27,400 mouse cDNAs, which were hybridized with fluorescent probes generated from mRNAs of T. cruzi infected and uninfected hearts. Differentially expressed genes were sorted according to their normalized expression patterns and functional groups including those involved in transcription, intracellular transport, structure/junction/adhesion or extracellular matrix, signaling, host defense, energetics, metabolism, cell shape and death. The regulated genes are interpreted in the pathogenesis of chagasic heart disease. PMID- 12910414 TI - Seasonal occurrence of the tapeworm Proteocephalus longicollis and its transmission from copepod intermediate host to fish. AB - Seasonal occurrence in terms of prevalence, intensity of infection, abundance and density of the tapeworm Proteocephalus longicollis (Zeder, 1800) and its transmission between its intermediate host (Cyclops abyssorum prealpinus) and definitive host (common whitefish, Coregonus lavaretus) in Lake Annecy, an oligotrophic lake in the western part of the Alps, France, were studied in the period of 1998-2000. A copepod Cyclops abyssorum prealpinus (Kiefer, 1939), the dominant species among planktonic crustaceans in the lake, served as the only intermediate host for this parasite. Infection with plerocercoids was higher in adult copepods (predominantly females) than in copepodite stages IV and V. The prevalence rate of 25% found in C. abyssorum prealpinus females in June 1998 represents a unique infection rate of intermediate hosts with fish tapeworms in natural conditions. The final host, the common whitefish Coregonus lavaretus (L.), was heavily infected with P. longicollis throughout the year (prevalence 90%; mean abundance 40.3; maximum intensity of infection more than 500 tapeworms per fish); immature tapeworms predominated in all samples (P<0.01). Transmission of tapeworm larvae from copepods to the common whitefish took place most intensively in summer and autumn, and depended on seasonal changes in the density of the C. abyssorum prealpinus population, infection of this copepod with plerocercoids and their density in the lake. In addition, transmission efficiency also seems to be determined by the longevity of tapeworm larvae in the intermediate host, timing of predation of the fish host and rapid development of the parasite within this host during the summer period. Overall transmission potential of P. longicollis was low, with only about 9% of juvenile specimens reaching maturity in common whitefish. PMID- 12910415 TI - Preliminary study on sex-related inflammatory reactions in mice infected with Schistosoma mansoni. AB - The aim of this study was to investigate the contribution of the sex of both the parasite and the host to the inflammatory response induced in unisexual infections of Schistosoma mansoni in mice. Organ weight, cell count and the delayed type hypersensitivity reaction were used as tools in this comparative study. The inflammatory reactions differed as a function of the sex of both the host and the parasite. Female mice showed a stronger inflammatory reaction to schistosome infection than males, while male schistosomes induced a stronger inflammatory response compared to females. The host-related differences in the inflammatory reaction may reflect differences in the factors affecting the immune defence of male and female mice. The differences in the inflammatory response induced by the parasite are discussed in terms of the quantity and quality of antigens among male and female worms. PMID- 12910416 TI - Study of specific IgG subclass antibodies for diagnosis of Gnathostoma spinigerum. AB - Gnathostoma spinigerum infection is endemic in Thailand and many Asian countries. Current diagnosis is the skin test and enzyme-linked immunosorbent assay (ELISA) for IgG antibody against the G. spinigerum third-stage larvae (L3), but cross reactivity is common. We evaluated the sensitivity and specificity of anti-G. spinigerum L3 IgG subclass antibodies for diagnosis of 43 patients with gnathostomiasis. The majority of patients with gnathostomiasis (91%) had eosinophilia. While the anti-G. spinigerum L3 IgG1 antibody provided the highest sensitivity (98%), the anti-G. spinigerum L3 IgG2 antibody had the highest specificity (88%). The ELISA that detected anti-G. spinigerum L3 IgG1 antibody could be a reliable laboratory screening test, while anti-G. spinigerum L3 IgG2 antibody could be used to confirm the diagnosis. PMID- 12910417 TI - The effect of the microtubule inhibitor tubulozole-C on the tegument of triclabendazole-susceptible and triclabendazole-resistant Fasciola hepatica. AB - The benzimidazole derivative, triclabendazole is the main drug used against Fasciola hepatica, although its precise mode of action remains to be fully determined. Previous studies have suggested that triclabendazole acts as a microtubule inhibitor in the same manner as tubulozole-C. Consequently, flukes from triclabendazole-susceptible and triclabendazole-resistant isolates were treated with tubulozole-C (1x10(-6 )M) in vitro and changes in tegumental morphology and tubulin distribution within the tegument were monitored by scanning and transmission electron microscopy, together with tubulin immunocytochemistry. Tubulozole-C caused severe disruption within the tegument of triclabendazole-susceptible flukes. Tubulin immunoreactivity diminished within the tegumental syncytium of the triclabendazole-susceptible flukes following treatment with tubulozole-C. In contrast, tubulozole-C caused only minor disruption of the tegument in triclabendazole-resistant F. hepatica and did not significantly alter the pattern of tubulin immunostaining within the tegumental syncytium. The results of the present study indicate that tubulozole-C and triclabendazole share the same target molecule and that triclabendazole-resistant flukes are also cross-resistant to tubulozole-C. PMID- 12910418 TI - Dihydropyrimidine dehydrogenase in normal and malignant endometrium: relationship with cell proliferation and thymidine phosphorylase. AB - Dihydropyrimidine dehydrogenase (DPD) is a pyrimidine salvage enzyme responsible for degradation of thymine, which is produced from thymidine by thymidine phosphorylase (TP). Our purpose was to determine the relationship between DPD, cell proliferation and TP expression in human endometrium. We examined DPD gene expression using reverse transcription-polymerase chain reaction, DPD protein levels using enzyme-linked immunosorbent assay, and DPD protein localization using immunohistochemistry in 58 normal endometria and 28 endometrial cancers. DPD gene expression was then related to the proliferating cell nuclear antigen index and to TP gene expression. DPD gene expression, which was correlated with DPD protein level, was relatively stable throughout various menstrual phases but was significantly elevated in postmenopausal status. It was significantly lower in endometrial cancer than in normal endometrium. Localization analysis revealed that DPD protein was located primarily in epithelial cells, but was also present in stromal cells. DPD gene expression correlated inversely with the PCNA index. TP gene expression pattern contrasted with that of DPD in postmenopausal and malignant endometrium. A high ratio of TP to DPD gene expression was significantly more frequent in endometrial cancer than in normal endometrium in any menstrual phase. DPD may act cooperatively with TP to affect cell function by maintaining the pyrimidine nucleotide pool balance in normal and malignant endometrium. PMID- 12910419 TI - Ovarian sex cord-stromal tumors--a clinicopathological study of 72 cases from the Kiel Pediatric Tumor Registry. AB - We analyzed 72 patients with ovarian sex cord-stromal tumors (OSCST) registered at the German Pediatric Tumor Registry in Kiel over a 20-year period. Juvenile granulosa cell tumors (JGCT, n=48) were the most frequent histological subtype. In addition, there were 14 Sertoli-Leydig cell tumors, 5 sclerosing stromal tumors, 2 sex cord tumors with annular tubules, 2 thecomas and 1 steroid cell tumor. Stage according to FIGO (International Federation of Gynecologists and Obstetricians) was Ia in 39 patients, Ic in 17 patients, II in 3 patients and III in 1 patient (60 patients with complete data). Compared with adult granulosa cell tumors, JGCT showed pronounced mitotic activity [mean 9.8 mitoses/10 high power field (HPF)], which was significantly higher than in other histological subtypes (2.7/10 HPF, P=0.001). Immunohistochemical analysis revealed frequent coexpression of vimentin (positive in 52/52 examined tumors), cytokeratin (27/33), and inhibin (19/20). Of patients, 12 with Ic or higher stage tumors received adjuvant cisplatinum-based chemotherapy. Event-free survival at 10 years was 0.88 +/- 0.05 (38/43 patients with follow-up data). Outcome significantly correlated with stage and mitotic activity (<20 versus > or =20 mitoses/10 HPF: event-free survival 1.0 versus 0.48 +/- 0.05, P=0.0001). In conclusion, this analysis confirms that the majority of patients with OSCST present at low tumor stage and that prognosis in these patients is excellent. Refractory tumors are characterized by high proliferative activity. Therefore, histopathological evaluation substantially contributes to risk assessment in patients with OSCST and might be useful for therapy stratification in prospective therapeutic protocols. PMID- 12910420 TI - Evaluation of gastroesophageal reflux disease following various reconstructive procedures for a distal gastrectomy. AB - BACKGROUND AND AIMS: Some patients develop gastroesophageal reflux disease (GERD) after a distal gastrectomy. In these patients, the evaluation of GERD with 24-h pH monitoring could be difficult because they may have an insufficient amount of gastric acid. PATIENTS AND METHODS: To evaluate GERD following a distal gastrectomy, we retrospectively evaluated 38 patients postoperatively with an acid reflux test (ART), a barium study, endoscopy, and esophageal manometry. Three reconstructive procedures, Billroth I (B-I group: 14 patients), Billroth II (B-II group: 11 patients), and jejunal interposition (interposition group: 13 patients) were compared with respect to GERD and short- term operation results. RESULTS: Operation time and postoperative hospital stay were longest in the interposition group. Reflux symptoms were present in ten patients (26%). The ART, barium study and endoscopy demonstrated evidence of GERD in 22 (58%), 10 (26%) and 4 (11%) of the 38 patients, respectively. The frequency of a positive ART in the interposition group was significantly lower than in both the B-I and the B-II groups. The abdominal length of the lower esophageal sphincter in the interposition group was higher than that in the B-II group ( P<0.05). CONCLUSION: Although jejunal interposition required longer operation time and hospital stay, the lower esophageal sphincter function following jejunal interposition appears to be superior to that following a Billroth-I or Billroth-II procedure. PMID- 12910421 TI - Is male gender a risk factor for bile duct injury during laparoscopic cholecystectomy? AB - BACKGROUND: Since its introduction in the late 1980s laparoscopic cholecystectomy has become the treatment of choice for gallstone disease. Unfortunately, the rate of iatrogenic biliary duct injuries (BDIs) has at least doubled after the adoption of the laparoscopic method. Population-based studies reporting the distribution of laparoscopic BDI patients according to gender and the severity of the BDI are mostly lacking. The purpose of the present study was to analyze the BDIs sustained during laparoscopic cholecystectomy in and around Turku University Central Hospital, with a special reference to the distribution of patients according to gender and the severity of the BDI. PATIENTS AND METHODS: A total of 3,736 laparoscopic cholecystectomies (2,627 female patients, 1,109 male) was performed in and around Turku University Central Hospital from 1995 to 2002 (by the end of April). The number and severity of BDIs and the gender of BDI patients were recorded, and the risk of BDI during laparoscopic cholecystectomy was calculated for the total patient population and for both genders separately. RESULTS: The risk of BDI was 0.86% for the total patient population, 0.95% for female and 0.63% for male. The most conspicuous finding was that the female gender was predominant in the severe types of BDI. However, the risk of mild BDI seemed to be fairly equal in both genders. CONCLUSION: We conclude that female gender seems to be a risk factor for severe iatrogenic BDI during laparoscopic cholecystectomy. PMID- 12910422 TI - Prospective evaluation of ultrasound and colour duplex imaging for the assessment of surgical resectability of pancreatic tumours. AB - BACKGROUND: This study was performed to evaluate colour duplex imaging (CDI) for the assessment of resectability of pancreatic tumours (PTs). METHOD: From October 1998 to December 2001, 182 patients consecutively having surgery for a PT were enrolled in this prospective study. Extension of the tumour to large blood vessels and retroperitoneum, the detection of liver metastases, enlarged lymph nodes and peritoneal carcinomatosis were defined as criteria for assessment. The patients were grouped into three classes of resectability: US-TU 1 = resectable/potentially curable, US-TU 2 = questionably resectable/curable, and US TU 3 = non-resectable/not curable. CDI was performed by five different examiners. Results were compared with intra-operative findings. RESULTS: Using CDI, we classified 85 (46.7%) tumours as resectable, 64 (35.2%) as non-resectable, and 33 (18.1%) as questionably resectable. Overall, 46.2% ( n=84) were found to be resectable, and 53.8% ( n=98) to be non-resectable, intra-operatively. A correlation between CDI classification and intra-operative findings was found in 138 of 149 cases (92.6%) (sensitivity 88.4%, specificity 96.3%). With regard to the complete oncological status (local extension, metastases, lymph-node staging and peritoneal carcinomatosis), a sensitivity of 77.2% and specificity of 95.7% were found. Non-correlated findings were likely attributed to missing small liver metastases, peritoneal carcinomatosis without ascites, and on difficulties in the assessment of enlarged lymph nodes concerning tumour infiltration. CONCLUSION: The use of CDI in evaluation of PTs may provide valuable pre-operative assessment of surgical resectability and may be performed in the clinical setting. PMID- 12910424 TI - [Palliative laser therapy of colorectal cancer with sustained local tumor removal]. AB - Laser photocoagulation is an established palliative therapeutic method in advanced colorectal cancer. In most cases tumor obstruction of the colonic lumen and symptoms like bleeding, secretion and diarrhoea can be improved or prevented. Even curative tumor therapy of some patients has been reported, particularly in cases with superficial growth of the tumor. We report about an elderly female patient with locally advanced invasive carcinoma of the rectosigmoidal junction. This tumor and a polypoid carcinoma of the sigmoid, possibly in combination with snare resection, were persistently removed by laser photocoagulation. PMID- 12910423 TI - [Endoscopic Therapy of Early Colorectal Cancer (pT1) - A Prospective Study]. AB - INTRODUCTION: In a prospective study initiated in 1982, we have been investigating the question as to whether - and if so, which - pT1 carcinomas of the colorectum can be treated exclusively via the endoscope. METHOD: In the period between February 1, 1982 and April 30, 2001, a total of 5,470 polyps were removed endoscopically at the Medical Department I of the Klinikum Ludwigsburg. Among these lesions, a total of 144 (2.6 %) pT1 carcinomas were found in 141 patients. We were able to follow 120 patients with 123 pT1 carcinomas over a mean follow-up period of 46 months (range: 1-60). In low-risk situations (definitive removal in healthy tissue, G1-G2, no lymphatic involvement), endoscopic treatment alone usually represented sufficient treatment. In high-risk cases (removal in healthy tissue uncertain or negative, and/or lymphatic vessel involvement, and/or G3/G4), subsequent surgical resection was carried out. RESULTS: 64 cases were classified as high-risk, 59 as low-risk. Nevertheless, 9 patients with 10 low risk carcinomas were submitted to surgery (young age, patient's own request). In none of these 10 cases was residual tumour or lymph node metastasis detected in the surgical specimen. 47 patients with 49 low-risk carcinomas were treated solely by endoscopic polypectomy using the diathermy snare, and 45 patients with 47 carcinomas remained recurrence-free during the follow-up period. In a single case, a local recurrence was detected 2 months after polypectomy and underwent curative resection. In another case, peritoneal carcinosis with tumour infiltrating into the colon developed 8 months after initial treatment; this, however, was most probably a recurrence of a previously operated carcinoma of the uterus. Among the high-risk cases, 10 were not submitted to surgery on account of advanced age and/or rejection of an operation by the patient; all remained recurrence-free. Among the surgically treated high-risk carcinomas, 3 surgical specimens contained residual tumour, while 2 revealed a lymph node metastasis. In our group of patients, no tumour-related mortality was seen among endoscopically treated patients. DISCUSSION: In the light of the fact that the reported mortality rate associated with open surgery for colorectal carcinoma is 3 % as compared with about 1 % risk of lymph node metastasis and 0,1 % mortality rate for the endoscopic modality, endoscopic removal of a pT1 tumour in a low-risk situation followed by appropriate surveillance can be considered as adequate treatment. PMID- 12910425 TI - [Angiomyolipoma of the colon - case report and review of the literature]. AB - Angiomyolipoma is one of the benign hamartomas that is found sporadically or associated with tuberous sclerosis. It is a rare soft tissue tumor involving mostly the kidneys, sometimes other visceral organs. The tumor is composed of smooth muscle cells, adipocytes and small sized hyalinized vessels. We present the case of a 74-year-old man with a bifocal angiomyolipoma of the colon. This diagnosis was confirmed at surgery, where a partial colectomy was performed. Histologic examination disclosed the tumor. The patient had no signs of family history of tuberous sclerosis. Extrarenal angiomyolipoma is rare and this may be the first report of bifocal colonic angiomyolipoma. PMID- 12910426 TI - [Secondary and primary prophylaxis of gastropathy associated with nonsteroidal antiinflammatory drugs or low-dose-aspirin: a review based on four clinical scenarios]. AB - Based on current references four clinical scenarios were discussed and different management strategies were compared for secondary and primary prophylaxis of ulcer or peptic ulcer bleeding under continuous therapy with non-steroidal antiinflammatory drugs (NSAID) or low-dose-aspirin, for H.pylori-positive and H.pylori-negative patients. Used as secondary prophylaxis eradication alone is insufficient in preventing recurrent peptic ulcer or recurrent ulcer bleeding for H.pylori-positive patients who continue to take unselective NSAIDs. Maintenance therapy with PPIs or switching from nonselective NSAID to COX-2-inhibitors is required after eradication of H.pylori or primary H.pylori-negative patients. Further evaluation is needed of what kind of secondary prophylaxis - maintenance therapy with PPI or switching to COX-2-inhibitor - is more (cost-)effective. It is sufficient to use eradication of H.pylori alone as secondary prophylaxis in preventing recurrent peptic ulcer or recurrent ulcer bleeding for H.pylori positive patients, who continue to take low-dose-aspirin. Maintenance therapy with PPI is not generally required. However it can be considered for patients with increased risk for gastrointestinal complications (previous history of peptic ulcer, age over 65 years, concomitant use of corticosteroids, anticoagulants or individual NSAID with higher risk for gastrointestinal complications, serious cardiovascular disease). Switching from low-dose-aspirin to clopidogrel is not required. Used as primary prophylaxis in preventing peptic ulcer or ulcer bleeding before starting long-term therapy with NSAIDs, COX-2 inhibitors or unselective NSAIDs concomitant with PPIs are recommended for patients with increased risk for gastrointestinal complications. Patients starting long-term therapy with unselective NSAIDs should be screened for H.pylori and eradicated. There are no valid data supporting screening for H.pylori and eradication for patients starting long-term therapy with low-dose aspirin. Further studies are needed to evaluate a possible benefit for patients with increased risk for gastrointestinal complications. PMID- 12910427 TI - [Surgical and medical therapy for gastroesophageal reflux disease. Consensus of pro/contra clinical debate]. PMID- 12910428 TI - [Specific localisation of human cytomegalovirus nucleic acids and proteins in human colorectal cancer]. PMID- 12910429 TI - [Celecoxib as well as Diclofenac in Combination with Omeprazole can Prevent the Recurrence of NSAIDs-Induced Gastrointestinal Bleeding]. PMID- 12910431 TI - [Review "Kava, kavapyrones and toxic liver injury" by R Teschke, Hanau Zeitschrift fur Gastroenterologie 2003, 41:395-404]. PMID- 12910433 TI - The Kleine-Levin syndrome - effects of treatment with lithium -. AB - Kleine-Levin syndrome (KLS) is a rare disorder which affects mainly adolescents. Periods of extreme somnolence alternate with megaphagia, psychomental changes and behavioural symptoms. The cause and pathogenesis of KLS remains unknown. Several treatments have been tried and recently lithium has been proposed for a prophylactic use in single cases. In view of the rarity of KLS, long-term results of lithium therapy have not been described yet. We report the clinical course of five adolescents with KLS who were treated with lithium. All patients showed significant EEG and polysomnographic changes during the episodes and had normal results in the interval. All patients had relapses while being treated with lithium. But episodes of hypersomnia under lithium therapy were shorter and monosymptomatic with lack of behavioural symptoms. Statistical modelling showed that the risk for a relapsing episode under maintenance of lithium drops per months of therapy from 100 % to 93 %, and furthermore that the maintenance of lithium shortens the mean duration of episodes to 19 %. No severe side effects were observed. In conclusion, in KLS with a high frequency of episodes and severe behavioural changes lithium may become a treatment option. PMID- 12910434 TI - Glatiramer acetate treatment in patients with childhood and juvenile onset multiple sclerosis. AB - BACKGROUND: Recent data indicate that in multiple sclerosis disease onset before the age of 16 is more common than previously assumed. However, current therapeutic options are limited to the treatment of acute attacks in these patients. Glatiramer acetate has been successfully applied in adults with relapsing-remitting multiple sclerosis, but there are no data available about the use of glatiramer acetate in childhood and juvenile onset multiple sclerosis. METHODS: Seven patients with relapsing-remitting multiple sclerosis and disease onset between 9 and 16 years of age were treated with daily subcutaneous injection of 20 mg glatiramer acetate (Copaxone). Patients were followed for 24 months. Treatment was initiated in all patients before the age of 18. RESULTS: The use of glatiramer acetate in our cohort of early onset multiple sclerosis patients was safe and well tolerated. Two out of seven patients remained relapse free during the two year period. Clinical disability as measured by the EDSS remained stable in three out of seven patients. CONCLUSION: Due to the small number of patients the efficacy of the treatment has to be interpreted with caution. However, there might be a more pronounced treatment benefit in patients with low disability at treatment initiation and early treatment onset. PMID- 12910435 TI - Degree of hypomyelination and magnetic resonance spectroscopy findings in patients with Pelizaeus Merzbacher phenotype. AB - As only 10 - 30 % of patients with a Pelizaeus Merzbacher disease (PMD) phenotype carry mutations of the proteolipid protein (PLP) gene, we were interested if the degree and time-dependent progression of abnormal MRI and MRS findings would discriminate patients with mutations of the PLP gene (Pelizaeus Merzbacher disease, PMD) from patients without a defect of the PLP gene (Pelizaeus Merzbacher-like disease, PMLD). For a standardised intraindividual follow-up and for comparison of the degree of hypomyelination, we have applied a newly developed semiquantitative myelination score on a total of 18 MRI series of 4 PMD and 4 PMLD patients. We found severe hypomyelination (< 50 % of normal) in 2 PMD and in 2 PMLD patients, moderate hypomyelination (< 75 % of normal) in 2 PMD and mild hypomyelination (> 75 % of normal) in 2 PMLD patients. Our score revealed a clear correlation between the degree of hypomyelination and the severity of clinical handicap in PMD but not in PMLD patients. MRS showed heterogeneous cerebral metabolite patterns in both patient groups and seems to reflect a mixture of unspecific changes due to primary hypomyelination and secondary gliosis and demyelination. Neither by MRI nor by MRS were patterns found that would allow differentiation between PMD and PMLD patients. PMID- 12910436 TI - NOx (nitrite/nitrate) in cerebral spinal fluids obtained from patients with influenza-associated encephalopathy. AB - It has become evident that the number of patients with a new type of influenza associated encephalopathy is increasing in Japan. Nitric oxide (NO), a simple free radical gas, elicits a wide range of physiological and pathophysiological effects. We measured the nitrite/nitrate (NO x ) levels in cerebral spinal fluid obtained from patients with influenza-associated encephalopathy in order to evaluate the correlation between the NO production and the process of influenza associated encephalopathy. Fifteen children were enrolled, aged from 1 to 9 years. As control we used 14 cerebral spinal fluids obtained from patients with urinary tract infection, respiratory infection or mumps meningitis without any sequela. NO 3 in influenza-associated encephalopathy was significantly higher than that of control group. On the other hand NO 2 was not significantly higher than that of control group. In particular, 4 out of 5 fatal cases revealed high NO 2 or NO 3. One case having normal levels in NO 2 and NO 3 showed that NH 3 was high. These results revealed that NO plays a role in influenza-associated encephalopathy through indirect effects of NO. PMID- 12910437 TI - Interleukin-6 and oligoclonal IgG synthesis in children with acute disseminated encephalomyelitis. AB - Interleukin-6 (IL-6) has a number of roles including recruitment of T lymphocytes and differentiation of B lymphocytes into IgG-secreting plasma cells. Furthermore, IL-6 is a neuropoietic cytokine with effects on neuronal differentiation, function and survival. We studied IL-6 concentrations in children with acute disseminated encephalomyelitis (ADEM; n = 14), and compared the values with those obtained from control patients with other inflammatory (OIND; n = 13) and non-inflammatory (NIND; n = 10) neurological disorders. Patients with ADEM had a significantly increased CSF IL-6 concentration compared with both OIND and NIND groups ( P < 0.01). Serum IL-6 was also increased in the ADEM group compared with the OIND group ( P < 0.05). CSF: serum IL-6 ratios were significantly increased in the ADEM group compared with the NIND group ( P < 0.05), suggesting an intrathecal production of IL-6 rather than its passive transfer across the blood-brain barrier alone. In ADEM, there was a significant correlation between an increased CSF IL-6 and an identical pattern of oligoclonal IgG synthesis in both serum and CSF ( P < 0.05). These results would suggest a role for IL-6 in the pathology of ADEM, and a possible direct link between an increased IL-6 and a proliferation of B lymphocytes with consequent IgG production. PMID- 12910438 TI - Familial bilateral medial parietooccipital band heterotopia not related to DCX or LIS1 gene defects. AB - A father and his daughter displayed strictly similar focal brain dysplasia at MR examination, characterized by regional medial posterior laminar sub-cortical grey matter heterotopia. To our knowledge, no family presenting such anomalies has yet been described. LIS1 and DCX gene defects were excluded. Collecting patients with such inherited dysplasia should improve our knowledge of the genetic basis of cortical malformations. PMID- 12910439 TI - Focal neuropathy in children with critical illness. AB - Four children are described who developed focal lower limb nerve palsies following critical illness. Two had clinical and/or neurophysiological evidence of simultaneous generalised critical illness polyneuropathy. The diagnosis was delayed in three patients due to the presence of central motor abnormalities and slow motor recovery. Follow-up from seven months to three years showed minimal or no recovery. In three, a vasculitic skin infarct, compartment syndrome and focal myositis could have caused nerve compression. We suggest that in critical illness peripheral nerves have an increased susceptibility to damage by local pressure. PMID- 12910440 TI - An unusual case of benign reflex myoclonic epilepsy of infancy. AB - A previously healthy one-year-old boy, the youngest child of unrelated parents, presented with a four-week history of episodes of myoclonus triggered only by tactile stimulation to his head. There had been no loss of developmental skills. The electroencephalogram (EEG) revealed generalised polyspike wave activity both with and without clinical correlate. The infant was started on sodium valproate, which resulted in cessation of the myoclonic episodes one week after starting therapy. At subsequent follow-up (at 18 months) the infant was seizure free and a repeat EEG was normal. This case of non-progressive reflex myoclonic epilepsy of infancy triggered only by head tapping (and not by acoustic stimuli) is an extremely rare phenomenon. Reflex myoclonic epilepsy of infancy represents a distinct subtype of myoclonic epilepsy in infancy. It should be considered as an age-dependent idiopathic generalised epileptic syndrome with an apparently good prognosis. PMID- 12910441 TI - New pattern of brain MRI lesions in isolated complex I deficiency. AB - We describe a boy presenting at the end of the first year of life with severely delayed motor development and only mild mental retardation. Neurological examination revealed axial hypotonia, mild ataxia and pyramidal signs. Elevated lactate and protein in cerebrospinal fluid were the most prominent laboratory abnormalities. Brain MRI showed severe supratentorial white matter changes. Cerebellar white matter appeared normal whereas the signal of the atrophic cerebellar cortex was markedly increased. In vivo 1H-magnetic resonance spectroscopy of the parietooccipital white matter region showed a distinct resonance of lactate. By means of biochemical analysis of respiratory chain enzymes in fibroblasts, the diagnosis of an isolated complex I deficiency could be established in our patient. PMID- 12910442 TI - Cerebellar hypoplasia with heterotopic purkinje cells in the molecular layer and preservation of the granule layers associated with severe encephalopathy. A new entity? AB - We present the case of an infant girl, born to first cousins, with a clinical phenotype consisting of microcephaly, hypotonia, strabismus and severe psychomotor retardation. Magnetic resonance imaging (MRI) showed global cerebellar atrophy involving the vermis and both hemispheres. The patient's serum transferrin levels were consistently unremarkable. Cerebellar biopsy, performed at 13 months of age, revealed heterotopic Purkinje cells in the molecular layer, but preservation of the external and internal granular layers. To our knowledge, this histological pattern of cerebellar cortical disorganization has not been described previously. The consanguinity of the parents suggests an autosomal recessive inheritance. PMID- 12910444 TI - [What does the Wallenberg syndrome teach us?]. PMID- 12910443 TI - Low thymic output and reduced heterogeneity of alpha/beta, but not gamma/delta, T lymphocytes in infants with ataxia-telangiectasia. AB - Ataxia-telangiectasia, a genetic disease caused by the homozygous mutation of the ATM gene, is frequently associated to a deficit of humoral and cellular immune functions. A decreased thymic output and skewed T cell and B cell receptor repertoires have been recently described in children over 7 years of age and in adults with this disease and have been proposed as a possible explanation for the immunodeficiency. To understand whether T cell defects arise early in life as a consequence of ATM gene mutations, we analysed the extent of thymic function by measuring the number of naive T cells and by studying the heterogeneity of T cells by means of heteroduplex analysis, in two children less than 2 years old with a remarkable reduction of T cell count. We found that the thymic output is decreased in babies with ataxia-telangiectasia if compared with that observed in age-matched normal babies. The low production of new T cells is associated to a reduction of the diversity of alpha/beta, but not gamma/delta, T lymphocytes. Our data indicate that ATM mutation limits the generation of a wide alpha/beta T cell repertoire and this feature can be responsible for the immunodeficiency observed in ataxia-telangiectasia babies. PMID- 12910445 TI - [Classical crossed syndromes of the medulla oblongata. A historical and topodiagnostic discussion]. AB - Historical publications of the classical alternating medulla oblongata syndromes of Wallenberg, Babinski-Nageotte, Cestan-Chenais, Hughlings Jackson, Avellis, Schmidt, Dejerine, Spiller and Tapia were reviewed and critically analysed. We compare these descriptions with descriptions of the brainstem syndromes in well known modern German, English and Russian neurological textbooks. The anatomic basis and etiology of the alternating medullar syndromes, and the main publications relating to these syndromes were discussed. Causes of the inconsistencies of the modern and historical descriptions of these syndromes might be an ignorance of the historical references. Progress and development of the clinical neurology and neuroanatomy in the late twentieth century, however, has also lead to correction and perfection of some historical descriptions in the modern neurological literature. PMID- 12910446 TI - [Neurological complications of inflammatory intestinal diseases]. AB - A number of neurological symptoms have been described as complications of inflammatory bowel disorders such as coeliac disease, Whipple's disease, Crohn's disease and ulcerative colitis. The neurology of coeliac disease includes disorders of the central nervous system encompassing cerebellar ataxia, epilepsy, myoclonus, dementia and multifocal leukoencephalopathy. Peripheral neuropathies, of axonal and demyelinating types, and myopathies have also been reported. The pathomechanisms are more related to immunological-inflammatory processes than to a malresorptive vitamin deficiencies. While CNS symptoms of coeliac disease show a poor response to gluten restriction, peripheral symptoms may respond to elimination of gluten from the diet. There are few reports of a successful corticosteroid treatment in patients showing inflammatory findings in the CSF or bioptic material. Whipple's disease is caused by the gram-positive bacillus Tropheryma whipplei. Patients with a central nervous system manifestation of Whipple's disease may develop a variety of symptoms including dementia, supranuclear gaze palsy, movement disorders, hypothalamic dysfunction and myorhythmia. The CNS-infection is diagnosed by PCR of the CSF. Long-term antibiotic treatment is required, and relapses may occur after withdrawal of antimicrobial therapy. Crohn's disease and ulcerative colitis are complicated by various forms of polyneuropathies, and arterial and venous cerebrovascular diseases. In most cases with neurological complications of inflammatory gastrointestinal diseases an immune mediated inflammatory process is suspected to be the underlying pathomechanism. PMID- 12910447 TI - [The next generation of "atypical" antipsychotics: the role of positron emission tomography]. AB - Almost fifteen years of research with Positron Emission Tomography (PET) and Single Photon Emission Computed Tomography (SPECT) have led to a profound understanding of the relationships between antipsychotic doses and plasma levels on the one hand and occupancy of (striatal) D2 -like dopamine receptors on the other hand as well as with the associated clinical effects and side effects. Furthermore, with the development of clinically "atypical" antipsychotics PET studies helped to generate hypotheses regarding the essential pharmacological properties of this heterogeneous class of drugs. Possible mechanisms of action include combined D2 -/5-HT2 antagonism, preferential mesolimbic binding, and fast dissociation from the D2 -receptor. Our recently published PET study on the in vivo characterization of the partial dopamine receptor agonist, aripiprazole, suggests a novel mechanism of action, which leads to clinically "atypical" properties of an antipsychotic. Aripiprazole, of which the antipsychotic efficacy has been proven in various multicenter clinical trials, leads to almost complete saturation of D2 -like dopamine receptors at clinically used doses; however, the incidence of extrapyramidal side effects under aripiprazole is not higher than under placebo. PET like no other method is suitable to display in vivo a novel mechanism of "atypicality" of a new class of antipsychotics. PMID- 12910448 TI - [Diagnostics and treatment of acute stroke]. PMID- 12910449 TI - Investigation of the induced-fit mechanism and catalytic activity of the human cytomegalovirus protease homodimer via molecular dynamics simulations. AB - Human cytomegalovirus (HCMV) is a highly species-specific DNA virus infecting up to 80% of the general population. The viral genome contains the open reading frame UL80, which encodes the full-length 80 kDa HCMV serine protease and its substrate. Full-length HCMV protease is composed of an N-terminal 256-amino-acid proteolytic domain, called assemblin, a linker region, and a C-terminal structural domain, the assembly protein precursor. Biochemical studies have shown that dimerization activates assemblin because of an induced stabilization of the oxyanion hole (Arg166). Thus, we performed here molecular dynamics (MD) simulations on HCMV protease models to study the induced-fit mechanism of the enzyme upon the binding of substrates and peptidyl inhibitors, and structural and energetic factors that are responsible for the catalytic activity of the enzyme dimer. Long and stable trajectories were obtained for the models of the monomeric and dimeric states, free in solution and bound to a peptidyl-activated carbonyl inhibitor, with very good agreement between theoretical and experimental results. Our results suggest that HCMV protease is indeed a novel example of serine protease that operates by an induced-fit mechanism. Also, in agreement with mutagenesis studies, our MD simulations suggest that the dimeric form is necessary to activate the enzyme because of an induced stabilization of the oxyanion hole. PMID- 12910450 TI - Discrimination of native loop conformations in membrane proteins: decoy library design and evaluation of effective energy scoring functions. AB - The recent determination of crystal structures for several important membrane proteins opens the way for comparative modeling of their membrane-spanning regions. However, the ability to predict correctly the structures of loop regions, which may be critical, for example, in ligand binding, remains a considerable challenge. To meet this challenge, accurate scoring methods have to discriminate between candidate conformations of an unknown loop structure. Some success in loop prediction has been reported for globular proteins; however, the proximity of membrane protein loops to the lipid bilayer casts doubt on the applicability of the same scoring methods to this problem. In this work, we develop "decoy libraries" of non-native folds generated, using the structures of two membrane proteins, with molecular dynamics and Monte Carlo techniques over a range of temperatures. We introduce a new approach for decoy library generation by constructing a flat distribution of conformations covering a wide range of Calpha-root-mean-square deviation (RMSD) from the native structure; this removes possible bias in subsequent scoring stages. We then score these decoy conformations with effective energy functions, using increasingly more cpu intensive implicit solvent models, including (1) simple Coulombic electrostatics with constant or distance-dependent dielectrics; (2) atomic solvation parameters; (3) the effective energy function (EEF1) of Lazaridis and Karplus; (4) generalized Born/Analytical Continuum Solvent; and (5) finite-difference Poisson Boltzmann energy functions. We show that distinction of native-like membrane protein loops may be achieved using effective energies with the assumption of a homogenous environment; thus, the absence of the adjacent lipid bilayer does not affect the scoring ability. In particular, the Analytical Continuum Solvent and finite-difference Poisson-Boltzmann energy functions are seen to be the most powerful scoring functions. Interestingly, the use of the uncharged states of ionizable sidechains is shown to aid prediction, particularly for the simplest energy functions. PMID- 12910451 TI - Simple two-state protein folding kinetics requires near-levinthal thermodynamic cooperativity. AB - Simple two-state folding kinetics of many small single-domain proteins are characterized by chevron plots with linear folding and unfolding arms consistent with an apparent two-state description of equilibrium thermodynamics. This phenomenon is hereby recognized as a nontrivial heteropolymer property capable of providing fundamental insight into protein energetics. Many current protein chain models, including common lattice and continuum Go models with explicit native biases, fail to reproduce this generic protein property. Here we show that simple two-state kinetics is obtainable from models with a cooperative interplay between core burial and local conformational propensities or an extra strongly favorable energy for the native structure. These predictions suggest that intramolecular recognition in real two-state proteins is more specific than that envisioned by common Go-like constructs with pairwise additive energies. The many-body interactions in the present kinetically two-state models lead to high thermodynamic cooperativity as measured by their van't Hoff to calorimetric enthalpy ratios, implying that the native and denatured conformational populations are well separated in enthalpy by a high free-energy barrier. It has been observed experimentally that deviations from Arrhenius behavior are often more severe for folding than for unfolding. This asymmetry may be rationalized by one of the present modeling scenarios if the effective many-body cooperative interactions stabilizing the native structure against unfolding is less dependent on temperature than the interactions that drive the folding kinetics. PMID- 12910452 TI - Contact order dependent protein folding rates: kinetic consequences of a cooperative interplay between favorable nonlocal interactions and local conformational preferences. AB - Physical mechanisms underlying the empirical correlation between relative contact order (CO) and folding rate among naturally occurring small single-domain proteins are investigated by evaluating postulated interaction schemes for a set of three-dimensional 27mer lattice protein models with 97 different CO values. Many-body interactions are constructed such that contact energies become more favorable when short chain segments sequentially adjacent to the contacting residues adopt native-like conformations. At a given interaction strength, this scheme leads to folding rates that are logarithmically well correlated with CO (correlation coefficient r = 0.914) and span more than 2.5 orders of magnitude, whereas folding rates of the corresponding Go models with additive contact energies have much less logarithmic correlation with CO and span only approximately one order of magnitude. The present protein chain models also exhibit calorimetric cooperativity and linear chevron plots similar to that observed experimentally for proteins with apparent simple two-state folding/unfolding kinetics. Thus, our findings suggest that CO-dependent folding rates of real proteins may arise partly from a significant positive coupling between nonlocal contact favorabilities and local conformational preferences. PMID- 12910453 TI - A left-handed alpha-helix containing both L- and D-amino acids: the solution structure of the antimicrobial lipodepsipeptide tolaasin. AB - The 18-amino acid cytolytic lipodepsipeptide tolaasin, produced in culture by virulent strains of Pseudomonas tolaasii, is the causal agent of the brown blotch disease of the cultivated mushroom. Tolaasin has a sequence of D-amino acids in its N-terminal region, then alternates L- and D-amino acids, and bears a C terminal lactone macrocycle composed of 5-residues. The solution structure of tolaasin in sodium dodecyl sulfate was studied by 2D-NMR spectroscopy and molecular dynamics simulated annealing calculations. Tolaasin forms an amphipathic left-handed alpha-helix in the regionDPro2-DalloThr14 comprising the sequence of seven D-amino acids and the adjacent L-D-L-D-D-region. To the best of our knowledge, this is the first recognized example of a left-handed alpha-helix including both D- and L-amino acids. The lactone macrocycle adopts a "boat-like" conformation and is shifted from the helical axis as to form a "golf-club" overall conformation. These structural features will be of importance in understanding, and preventing, tolaasin's role in the bacterial colonization of the host plant, and its toxic action on cells. Furthermore, the observed antimicrobial activity together with the potential resistance to enzymatic degradation and the increased antigenicity (both due to the presence of L- and D amino acids) strongly suggests for tolaasin a potential role as a template model for the design of new therapeutic antibacterial molecules. PMID- 12910454 TI - Identification of functionally conserved residues with the use of entropy variability plots. AB - We introduce sequence entropy-variability plots as a method of analyzing families of protein sequences, and demonstrate this for three well-known sequence families: globins, ras-like proteins, and serine-proteases. The location of an aligned residue position in the entropy-variability plot correlates with structural characteristics, and with known facts about the roles of individual amino acids in the function of these proteins. The large numbers of known sequences in these families allowed us to introduce new filtering methods for variability patterns. The results are discussed in terms of a simple evolutionary model for functional proteins. PMID- 12910455 TI - Sequence analysis reveals how G protein-coupled receptors transduce the signal to the G protein. AB - Sequence entropy-variability plots based on alignments of very large numbers of sequences-can indicate the location in proteins of the main active site and modulator sites. In the previous article in this issue, we applied this observation to a series of well-studied proteins and concluded that it was possible to detect most of the residues with a known functional role. Here, we apply the method to rhodopsin-like G protein-coupled receptors. Our conclusion is that G protein binding is the main evolutionary constraint on these receptors, and that other ligands, such as agonists, act as modulators. The activation of the receptors can be described as a simple, two-step process, and the residues involved in signal transduction can be identified. PMID- 12910456 TI - Spatial profiling of protein hydrophobicity: native vs. decoy structures. AB - A recent study of 30 soluble globular protein structures revealed a quasi invariant called the hydrophobic ratio. This invariant, which is the ratio of the distance at which the second order hydrophobic moment vanished to the distance at which the zero order moment vanished, was found to be 0.75 +/- 0.05 for 30 protein structures. This report first describes the results of the hydrophobic profiling of 5,387 non-redundant globular protein domains of the Protein Data Bank, which yields a hydrophobic ratio of 0.71 +/- 0.08. Then, a new hydrophobic score is defined based on the hydrophobic profiling to discriminate native-like proteins from decoy structures. This is tested on three widely used decoy sets, namely the Holm and Sander decoys, Park and Levitt decoys, and Baker decoys. Since the hydrophobic moment profiling characterizes a global feature and requires reasonably good statistics, this imposes a constraint upon the size of the protein structures in order to yield relatively smooth moment profiles. We show that even subject to the limitations of protein size (both Park & Levitt and Baker sets are small protein decoys), the hydrophobic moment profiling and hydrophobic score can provide useful information that should be complementary to the information provided by force field calculations. PMID- 12910457 TI - Flavors of protein disorder. AB - Intrinsically disordered proteins are characterized by long regions lacking 3-D structure in their native states, yet they have been so far associated with 28 distinguishable functions. Previous studies showed that protein predictors trained on disorder from one type of protein often achieve poor accuracy on disorder of proteins of a different type, thus indicating significant differences in sequence properties among disordered proteins. Important biological problems are identifying different types, or flavors, of disorder and examining their relationships with protein function. Innovative use of computational methods is needed in addressing these problems due to relative scarcity of experimental data and background knowledge related to protein disorder. We developed an algorithm that partitions protein disorder into flavors based on competition among increasing numbers of predictors, with prediction accuracy determining both the number of distinct predictors and the partitioning of the individual proteins. Using 145 variously characterized proteins with long (>30 amino acids) disordered regions, 3 flavors, called V, C, and S, were identified by this approach, with the V subset containing 52 segments and 7743 residues, C containing 39 segments and 3402 residues, and S containing 54 segments and 5752 residues. The V, C, and S flavors were distinguishable by amino acid compositions, sequence locations, and biological function. For the sequences in SwissProt and 28 genomes, their protein functions exhibit correlations with the commonness and usage of different disorder flavors, suggesting different flavor-function sets across these protein groups. Overall, the results herein support the flavor-function approach as a useful complement to structural genomics as a means for automatically assigning possible functions to sequences. PMID- 12910458 TI - Survey for g-proteins in the prokaryotic genomes: prediction of functional roles based on classification. AB - The members of the family of G-proteins are characterized by their ability to bind and hydrolyze guanosine triphosphate (GTP) to guanosine diphosphate (GDP). Despite a common biochemical function of GTP hydrolysis shared among the members of the family of G-proteins, they are associated with diverse biological roles. The current work describes the identification and detailed analysis of the putative G-proteins encoded in the completely sequenced prokaryotic genomes. Inferences on the biological roles of these G-proteins have been obtained by their classification into known functional subfamilies. We have identified 497 G proteins in 42 genomes. Seven small GTP-binding protein homologues have been identified in prokaryotes with at least two of the diagnostic sequence motifs of G-proteins conserved. The translation factors have the largest representation (234 sequences) and are found to be ubiquitous, which is consistent with their critical role in protein synthesis. The GTP_OBG subfamily comprises of 79 sequences in our dataset. A total of 177 sequences belong to the subfamily of GTPase of unknown function and 154 of these could be associated with domains of known functions such as cell cycle regulation and t-RNA modification. The large GTP-binding proteins and the alpha-subunit of heterotrimeric G-proteins are not detected in the genomes of the prokaryotes surveyed. PMID- 12910459 TI - How well can we predict native contacts in proteins based on decoy structures and their energies? AB - One strategy for ab initio protein structure prediction is to generate a large number of possible structures (decoys) and select the most fitting ones based on a scoring or free energy function. The conformational space of a protein is huge, and chances are rare that any heuristically generated structure will directly fall in the neighborhood of the native structure. It is desirable that, instead of being thrown away, the unfitting decoy structures can provide insights into native structures so prediction can be made progressively. First, we demonstrate that a recently parameterized physics-based effective free energy function based on the GROMOS96 force field and a generalized Born/surface area solvent model is, as several other physics-based and knowledge-based models, capable of distinguishing native structures from decoy structures for a number of widely used decoy databases. Second, we observe a substantial increase in correlations of the effective free energies with the degree of similarity between the decoys and the native structure, if the similarity is measured by the content of native inter-residue contacts in a decoy structure rather than its root-mean-square deviation from the native structure. Finally, we investigate the possibility of predicting native contacts based on the frequency of occurrence of contacts in decoy structures. For most proteins contained in the decoy databases, a meaningful amount of native contacts can be predicted based on plain frequencies of occurrence at a relatively high level of accuracy. Relative to using plain frequencies, overwhelming improvements in sensitivity of the predictions are observed for the 4_state_reduced decoy sets by applying energy-dependent weighting of decoy structures in determining the frequency. There, approximately 80% native contacts can be predicted at an accuracy of approximately 80% using energy-weighted frequencies. The sensitivity of the plain frequency approach is much lower (20% to 40%). Such improvements are, however, not observed for the other decoy databases. The rationalization and implications of the results are discussed. PMID- 12910460 TI - Improved protein-ligand docking using GOLD. AB - The Chemscore function was implemented as a scoring function for the protein ligand docking program GOLD, and its performance compared to the original Goldscore function and two consensus docking protocols, "Goldscore-CS" and "Chemscore-GS," in terms of docking accuracy, prediction of binding affinities, and speed. In the "Goldscore-CS" protocol, dockings produced with the Goldscore function are scored and ranked with the Chemscore function; in the "Chemscore-GS" protocol, dockings produced with the Chemscore function are scored and ranked with the Goldscore function. Comparisons were made for a "clean" set of 224 protein-ligand complexes, and for two subsets of this set, one for which the ligands are "drug-like," the other for which they are "fragment-like." For "drug like" and "fragment-like" ligands, the docking accuracies obtained with Chemscore and Goldscore functions are similar. For larger ligands, Goldscore gives superior results. Docking with the Chemscore function is up to three times faster than docking with the Goldscore function. Both combined docking protocols give significant improvements in docking accuracy over the use of the Goldscore or Chemscore function alone. "Goldscore-CS" gives success rates of up to 81% (top ranked GOLD solution within 2.0 A of the experimental binding mode) for the "clean list," but at the cost of long search times. For most virtual screening applications, "Chemscore-GS" seems optimal; search settings that give docking speeds of around 0.25-1.3 min/compound have success rates of about 78% for "drug like" compounds and 85% for "fragment-like" compounds. In terms of producing binding energy estimates, the Goldscore function appears to perform better than the Chemscore function and the two consensus protocols, particularly for faster search settings. Even at docking speeds of around 1-2 min/compound, the Goldscore function predicts binding energies with a standard deviation of approximately 10.5 kJ/mol. PMID- 12910461 TI - Structure, evolution, and inhibitor interaction of S-adenosyl-L-homocysteine hydrolase from Plasmodium falciparum. AB - S-adenosylhomocysteine hydrolase (SAHH) is a key regulator of S adenosylmethionine-dependent methylation reactions and an interesting pharmacologic target. We cloned the SAHH gene from Plasmodium falciparum (PfSAHH), with an amino acid sequence agreeing with that of the PlasmoDB genomic database. Even though the expressed recombinant enzyme, PfSAHH, could use 3-deaza adenosine (DZA) as an alternative substrate in contrast to the human SAHH, it has a unique inability to substitute 3-deaza-(+/-)aristeromycin (DZAri) for adenosine. Among the analogs of DZA, including neplanocin A, DZAri was the most potent inhibitor of the PfSAHH enzyme activity, with a K(i) of about 150 nM, whether Ado or DZA was used as a substrate. When the same DZA analogs were tested for their antimalarial activity, they also inhibited the in vitro growth of P. falciparum parasites potently. Homology-modeling analysis revealed that a single substitution (Thr60-Cys59) between the human and malarial PfSAHH, in an otherwise similar SAH-binding pocket, might account for the differential interactions with the nucleoside analogs. This subtle difference in the active site may be exploited in the development of novel drugs that selectively inhibit PfSAHH. We performed a comprehensive phylogenetic analysis of the SAHH superfamily and inferred that SAHH evolved in the common ancestor of Archaea and Eukaryota, and was subsequently horizontally transferred to Bacteria. Additionally, an analysis of the unusual and uncharacterized AHCYL1 family of the SAHH paralogs extant only in animals reveals striking divergence of its SAH-binding pocket and the loss of key conserved residues, thus suggesting an evolution of novel function(s). PMID- 12910464 TI - Characteristics of reproductive biology and proximate factors regulating seasonal breeding in captive golden-headed lion tamarins (Leontopithecus chrysomelas). AB - Reproduction is highly demanding in terms of energy expenditure, and the costs and benefits associated with postponing or investing in a reproductive effort are crucial determinants of an individual's fitness. Understanding the reproductive potential of a species under varying ecological conditions offers important insights into the dynamics of its social system. This study provides the first detailed analysis of the reproductive potential of wild- and captive-born golden headed lion tamarins (Leontopithecus chrysomelas) under captive conditions, based on studbook data compiled during 1984-2000. Litters produced by wild-born females breeding in captivity are similar in size to litters observed in the wild, but smaller than litters of captive-born females. The more stringent ecological conditions experienced by wild-born females during maturation may result in a lifelong effect on litter size. However, interbirth intervals are shorter for wild-born than captive-born females. The relatively smaller burden of infant care that results from having smaller litters may allow wild-born females to sustain the next pregnancy sooner. Reproduction in the Brazilian captive population is highly seasonal for both wild-born females and females born in captivity in Brazil. Changes in photoperiod over a year provide a proximate explanation for changes in the proportion of conceptions and births per month. Outside Brazil, breeding occurs year-round, and no clear birth peak is apparent. Information from field reports that could be used to relate this finding to ecological factors, such as resource availability, is unavailable. PMID- 12910462 TI - Crystal structure of a flavin-binding protein from Thermotoga maritima. PMID- 12910466 TI - Possible audience effect in Thomas langurs (Primates; Presbytis thomasi): an experimental study on male loud calls in response to a tiger model. AB - Recent research indicates that animal vocalizations can refer to objects in the outside world, and that call production may be controlled by the sender depending on the type of audience involved. Our research on wild male Thomas langurs (Primates: Presbytis thomasi) shows that these males call as a response to a tiger model when they are in a group, but do not call when they are solitary. This is the first experimental study on wild primates to demonstrate that the presence or absence of an audience influences calling behavior. Our results indicate that males in mixed-sex groups give more loud calls than solitary males when exposed to a predator model. This suggests that giving loud calls in response to predators is not purely a reflex and may be controlled in some way by the sender. PMID- 12910465 TI - Experimental field study of the relative costs and benefits to wild tamarins (Saguinus imperator and S. fuscicollis) of exploiting contestable food patches as single- and mixed-species troops. AB - Several species of tamarins form stable mixed-species troops in which groups of each species feed, forage, rest, and travel together during much of the year. Although the precise set of factors that facilitate this ecological relationship remains unclear, predator detection and foraging benefits are presumed to play a critical role in maintaining troop stability. In this work we present data from an experimental field study designed to examine how factors such as social dominance and within-patch foraging decisions affect the costs and benefits to tamarins of visiting feeding sites as single- and mixed-species troops. Our data indicate that when they exploited contestable food patches (sets of eight feeding platforms, two of which contained a 100-g banana), each tamarin species experienced foraging costs when they arrived as part of a mixed-species troop. These costs were found to be less severe for emperor tamarins because they were socially dominant to saddle-back tamarins and could displace them at feeding sites. We conclude that the foraging benefits to tamarins residing in mixed species troops are asymmetrical, and that at feeding sites in which the amount of food in a patch is insufficient to satiate all troop members, even minor differences in the timing of return to food patches and changes in troop cohesion have a measurable effect on the costs and benefits to participating tamarin species. PMID- 12910467 TI - Diurnal patterns of urinary steroid excretion in wild chimpanzees. AB - Urinary testosterone and cortisol concentrations were quantified in a large number of samples (>500) collected from wild male chimpanzees (n=11) over the course of 1 year. For both steroids, urinary concentrations were higher and more variable in the morning than in the afternoon. Urinary creatinine levels showed no such diurnal pattern. These patterns are consistent with studies of steroid excretion in humans and gorillas. This study emphasizes the importance of considering time of day as a confounding variable in field studies of primate endocrine function. It also suggests that if a small number of samples are to be used to characterize an individual's basal steroid levels, afternoon samples may be preferable because they show less intra-individual variability. PMID- 12910468 TI - Leaf-pile pulling: an unusual play pattern in wild chimpanzees. AB - Play in nonhuman animals has generally been viewed as being uniform among study sites. No studies have examined whether there are local variations in play. In this work we report an apparently locality-specific form of play that is basically solo locomotor play, but also has aspects of object play and social play. We describe this unusual "leaf-pile pulling" (LPL) pattern based on video footage of the chimpanzees of Mahale, Tanzania. Typically, when a party of chimpanzees moves in a procession down a slope in the dry season, a youngster will turn around and walk backward while raking many dry leaves with both hands. This activity accumulates many dry leaves while producing a lot of sound. After the player walks 1-15 m, he/she either turns around and walks forward or moves in a somersaulting fashion. The performer usually faces an individual that is immediately following him/her in the procession. The age of the performers ranges from 2 to 22 years, but 3-10 years are most typical. Compared to younger (< 8 years) individuals, older (> or = 8 years) individuals tend to cover longer distances during play, and to be more likely to play only on sloped surfaces and during travel. One of the authors (W.W.) has only seen the behavior exhibited by five different individuals on 10-15 occasions at Gombe. The behavior is exhibited less often at Gombe than at Mahale. To date, LPL has not been reported elsewhere. The absence of LPL at other sites may or may not be explained by environmental differences, such as differences in the availability of dead leaves and sloped terrain. Although the possibility that LPL is acquired by individual learning cannot be completely ruled out, the hypothesis that it is a tradition of the Mahale study group is more likely. This suggests that play may warrant more careful consideration in studies of nonhuman primate culture. PMID- 12910469 TI - Functional cellular biochemistry: a memorial to the late Dr. Joe Chayen. Meeting proceedings, 14 November 2002. PMID- 12910470 TI - Out of one eye: a life integrating cellular biochemistry and function. PMID- 12910471 TI - Messenger DNA in higher plants. AB - Evidence is presented that, as in animal and human cells, plant cells can release a newly-synthesized DNA which can freely circulate in the plants. This DNA enters cells and their nuclei where it may be integrated and be expressed so acting, apparently, as a messenger-DNA. PMID- 12910472 TI - Chromatin-associated sphingomyelin: metabolism in relation to cell function. AB - After the first histochemical demonstration by Chayen and Gahan of the presence of phospholipids and especially of sphingomyelin in chromatin, this became the object of long debate and of contradictory results. The general conclusion was that the presence of phospholipids may due to contamination during the isolation of chromatin. More recently the existence of a phospholipid chromatin fraction was confirmed by demonstrating that isolated hepatocyte nuclei, labelled by saturated and unsaturated radioiodination method, showed the presence of radioactivity only in the membrane and not in the isolated chromatin. The phospholipid composition showed an enrichment in sphingomyelin which increased during hepatocyte maturation or erythroleukemic cell differentiation induced by DMSO. A decrease in sphingomyelin was observed at the beginning of the S-phase in regenerating liver or in cultured proliferating cells. These changes were due to the presence of sphingomyelinase and sphingomyelin synthase in the chromatin, the activity of which paralleled the variation in sphingomyelin content. The sphingomyelin was co-localized with RNA as shown by biochemical and electron microscopy methods. Using bromo-uridine it was demonstrated that labelled RNA and sphingomyelin were present in actively transcribing nuclear regions. Isolated nuclear complexes after DNase and RNase digestion contained not only protein, but also RNA and sphingomyelin. After hydrolysis of sphingomyelin the RNAse-resistant RNA becomes RNAse sensitive. It can therefore be concluded that sphingomyelin and the related enzymes are present in the chromatin; sphingomyelin may have a role in RNA transcription protecting RNA by RNAse digestion before its transfer to the cytoplasm. PMID- 12910473 TI - Annexin 1: a paracrine/juxtacrine mediator of glucorticoid action in the neuroendocrine system. AB - Glucocorticoids (GCs) play an essential role in the maintenance of homeostasis. In normal circumstances their secretion is tightly regulated by a complex servo mechanism through which the steroids suppress the synthesis and release of ACTH and its hypothalamic releasing factors (CRH and AVP) and thereby reduce the positive drive to the adrenal cortex. The feedback actions of GCs on hormone release develop rapidly (within minutes), well before any changes in hormone synthesis are apparent. By using immunoneutralization, gene targeting and pharmacological strategies in in vivo and in vitro models, we have identified annexin 1, a Ca(2+)- and phospholipid-binding protein, as a key mediator of the early inhibitory actions of GCs on peptide release. This brief review outlines this work and describes molecular and cellular studies which have provided insight into the mechanism of annexin 1-dependent GC signalling in the neuroendocrine system. PMID- 12910474 TI - Investigation of the regulation of bone mass by mechanical loading: from quantitative cytochemistry to gene array. AB - From the 1860s to the early 1980s, the process that fitted bone architecture and mass to function had been investigated and characterized. It was known that increases in exercise were associated with increased bone mass, and that disuse caused osteopaenia, but the mechanisms by which those processes were regulated was not understood. The idea that osteocytes, the cells embedded in bone, were sensitive to the effects of mechanical loading was attractive, yet there was almost no experimental support for it, at least in part because the cells were considered inaccessible for study. In 1984, the techniques devised by Chayen and his co-workers were focused on this area. By analysis of the activity of the enzyme glucose 6-phosphate dehydrogenase in osteocytes in sections of avian bone that had been subjected to brief periods of applied mechanical loading, we showed for the first time that osteocytes could respond within a few minutes to mechanical stimulation. The lack of elevation of activity of other glycolytic enzymes led to the conclusion that this elevation was due to increased activity of the pentose shunt pathway, which was likely to be associated with increased production of reducing equivalents for biosynthesis, and ribose sugars for RNA synthesis. This was the first demonstration of an ability of osteocytes to respond to an external mechanical event and in effect provided a mechanistic link for the fundamental principle of what is known as Wolff's law of bone remodelling. These studies were dependent on several technical advances brought together in the Chayen Cellular Biology Laboratory at the Kennedy Institute. The ability to make cryosections of undecalcified bone, to perform cytochemical analysis of (soluble) enzyme activities by use of colloid stabilizers in the reaction medium, and finally to measure accurately the coloured reaction products by microdensitometry (which avoided optical heterogeneity errors) combined to provide a powerful way to explore bone cell function in situ. In the intervening years since then, similar studies have become routine, and the impact of molecular biological advances in hard tissues have remained dependent on techniques pioneered in the Chayen laboratory. During such studies, other advances have spun off, so that osteocyte gene expression has been analysed in samples taken from sections where the precise tissue characteristics were known, leading to advances in understanding of intercellular signalling mechanisms in bone by differential display, and the role of apoptosis in osteocytes in regulation of osteoclastic resorption. Still more recently, materials extracted from undecalcified sections have been used in gene array studies to discover new candidate genes with a role in the adaptive mechanism. Without Joe Chayen's involvement in this area, which now impacts on almost all bone biological science either directly or indirectly, our understanding of the pathophysiology of osteoporosis would have been very different. PMID- 12910475 TI - A cytochemical assay for osteoclast cathepsin K activity. AB - Cathepsin K is a member of the papain superfamily of cysteine proteases and plays a pivotal role in osteoclast-mediated bone resorption. This enzyme is an excellent target for antiresorptive therapies for osteopenic disorders such as osteoporosis.(1) Although isolated inhibitor studies on purified enzymes is required to discover potent and selective inhibitors of cathepsin K, a quantitative cytochemical assay(2) for cathepsin K would allow inhibitors to be tested on actual osteoclasts within sections of bone. Furthermore cathepsin K activity could be used to identify and analyse osteoclasts at definitive stages of their lifespan. A cytochemical assay is described that localizes osteoclast cathepsin K activity in unfixed, undecalcified cryostat sections of animal and human bone. PMID- 12910476 TI - Identifying and characterizing the joint cavity-forming cell. AB - For many years, a large body of circumstantial evidence supported the notion that the synovial membrane produced the hyaluronan-rich synovial fluid. A quantitative cytochemical technique for uridine-diphospho glucose dehydrogenase (UDPGD) activity established that fibroblast-like cells on the intimal surface of the synovial lining made a specific contribution to maintaining these glycosaminoglycan levels. Our studies have aimed to determine the mechanisms that control the attainment and persistence of this differentiated phenotype, and have recently focused on their appearance during joint cavity development in the embryonic limb; a process that is dependent upon skeletal movement. These in situ micro-biochemical studies have shown that cells bordering the presumptive joint cavity exhibit raised UDPGD activity, are associated with a matrix rich in hyaluronan and show immobilization-induced loss in such characteristics. Together with complimentary studies in adult joints, this suggests that mechanical stimuli promote the acquisition of this joint line-forming phenotype. For this reason our studies have attempted to identify the 'up-stream' mechano-dependent factors that control these events. Endothelial cells respond to mechanical stimuli by activating, via phosphorylation, mitogen activated protein kinase/extracellular signal-regulated kinase (MAPkinase/ERK). Using phospho-specific anti-ERK-1/2 antibodies we have shown that immunolabelling of developing limbs shows a clear joint line-selective activation during cavitation, with little if any labelling within neighbouring elements, and that this is abolished in immobilized limbs. In an attempt to facilitate the final mechanistic deciphering of these responses we have used an in vitro-based approach and found by Western blotting that active ERK-1/2 expression was increased in cultured articular surface cells following application of dynamic mechanical strain. Intriguingly, the use of a selective inhibitor (PD98059) of ERK activation by its classical activating kinase, Mek, to restrict such strain-induced increases, produced an enhanced strain-related increase in UDPGD mRNA expression. This suggests that mechano-dependent ERK activation serves a feedback regulatory role during differentiation of these cells. Whilst it is clear that these in vitro experiments serve a useful function, it is clear that they generally take little regard of the influence that might be provided by cell-cell and cell-matrix interactions within the developing limb's complex and dynamic environment and architecture. It is therefore imperative that we attempt to bridge the gap between the cell biology of such phenomena on the one hand, and the morphological approach to this same problem on the other. PMID- 12910477 TI - Cytochemical bioassays: every dog has its day. PMID- 12910479 TI - The relationship between total sialic acid levels and antioxidant status in the tadpoles of Bufo viridis and Rana ridibunda ridibunda. AB - Total sialic acid levels (TSA), antioxidant enzymes activities such as superoxide dismutase (SOD), catalase (CAT) and lipid peroxidation (LPO) levels were investigated during the developmental period in tadpoles of the predominantly terrestrial amphibian B. viridis and the predominantly aquatic amphibian R. r. ridibunda. Maximum TSA levels were observed in B. viridis and R. r. ridibunda at the fifth and third week of their development, respectively. SOD and CAT activity variations during development in B. viridis were higher than in R. r. ridibunda. Although SOD activity in B. viridis was higher than R. r. ridibunda at the eighth week, SOD activity increased 19.2-fold in R. r. ridibunda and 10.4-fold in B. viridis between the first and eighth week. CAT activity in R. r. ridibunda did not significantly change (p>0.001) until the fifth week then increased, whereas in B. viridis CAT increased after the third week. In contrast to the rise in the antioxidant enzyme activities, LPO levels tended to decrease during the developmental period. Levels of LPO showed a similar trend until the third week for both species. The minimum LPO levels in B. viridis and R. r. ridibunda were 23+/-1.2 and 146+/-7.3 nmol MDA g(-1) tissue, at the eighth week, respectively. While decreasing LPO levels correlated with increasing antioxidant enzyme activities, TSA tended to decrease after reaching a maximum point. PMID- 12910478 TI - Oxidized LDL induces transcription factor activator protein-1 in rat mesangial cells. AB - It has been shown that oxidized low-density lipoprotein (ox-LDL), through the activation of glomerular cells, stimulates pathobiological processes involved in monocyte infiltration into the mesangium. The underlying molecular mechanisms are not fully understood. The present study showed that ox-LDL strongly induced AP-1 binding activity in rat mesangial cells (RMCs) in a dose- and time-dependent manner, reaching the maximal activation at 250 microg ml(-1) within 24 h. The results from mobility shift assays and Western blotting analysis revealed that this AP-1 binding increase involved c-Jun, but not c-Fos. Moreover, this ox-LDL increased AP-1 binding was inhibited by several protein kinase (PK) inhibitors: the protein kinase C (PKC) inhibitor Bisindolylmaleimide I, the cAMP-dependent PK (PKA) inhibitor H89, and the tyrosine PK (PTK) inhibitor genistein. Protein phosphorylation represents mitogen-activated protein kinase (MAPK) activity. Therefore, we examined the role of ox-LDL on the activation of mesangial cell JNK/SAPK, the only recognized protein kinase that catalyses phosphorylation of c Jun. The incubation of mesangial cells with ox-LDL induced phosphorylation of JNK1/SAPK dose dependently, with the maximal response at 150 microg ml(-1). This study demonstrates that multiple kinase activities are involved in the mechanism of ox-LDL-induced AP-1 activation in mesangial cells, and ox-LDL stimulates AP-1 through JNK-c-Jun other than MEK-c-Fos signalling pathway. PMID- 12910480 TI - Analysis of intracellular distribution and apoptosis involvement of the Ufd1l gene product by over-expression studies. AB - UFD1L is the human homologue of the yeast ubiquitin fusion degradation 1 (Ufd1) gene and maps on chromosome 22q11.2 in the typically deleted region (TDR) for DiGeorge/velocardiofacial syndromes (DGS/VCFS). In yeast, Ufd1 protein is involved in a degradation pathway for ubiquitin fused products (UFD pathway). Several studies have demonstrated that Ufd1 is a component of the Cdc48-Ufd1-Npl4 multiprotein complex which is active in the recognition of several polyubiquitin tagged proteins and facilitates their presentation to the 26S proteasome for protein degradation or even more specific processing. The multiprotein complex Cdc48-Ufd-Npl4 is also active in mammalian cells. The biochemical role of UFD1L protein in human cells is unknown, even though the interaction between UFD1L and NPL4 proteins has been maintained. In order to clarify this issue, we examined the intracellular distribution of the protein in different mammalian cells and studied its involvement in the Fas and ceramide factors-mediated apoptotic pathways. We established that in mammalian cells, Ufd1l is localized around the nucleus and that it does not interfere with Fas-and ceramide-mediated apoptosis. PMID- 12910482 TI - Changes in adenosine deaminase activity in ageing cultured human cells and the role of zinc. AB - The level of adenosine deaminase (ADA; EC 3.5.4.4) was estimated at different passages in six confluent fibroblast cultures established from forearm skin biopsies of healthy adult normal volunteers. After determination of the zinc concentration in standard growth medium, ADA activity was estimated at different passages of subculture in media with different zinc concentrations. The results indicated that the specific activity of ADA in control confluent skin fibroblast cultures (passage 2) cultivated in standard growth medium containing 15.4 microM zinc (similar to that present in normal human plasma) was equal to 226.6+/-19.64 micromol min(-1) mg(-1) protein. The results showed that there were no significant changes in ADA specific activity in any of the control cultures as the zinc concentration of the medium was increased. To characterize the passage of subculture at which fibroblasts enter the ageing phase, three marker enzymes were assayed namely, phosphofructokinase, lactate dehydrogenase and glycogen phosphorylase. The result showed that the cells enter the ageing phase at passage 20 and beyond. Further investigation showed that ADA activity of serially subcultured confluent cultures cultivated in standard growth medium significantly dropped at passages 20, 25 and 30. ADA activity however was not significantly altered in cells at passage 2, 10 and 15 cultivated in standard growth medium and in the presence of higher zinc levels (23.1, 34.6, 53.8 and 73.1 microM). Furthermore there was significant lowering of ADA activities in cells at passages 20, 25 and 30 when cells were cultured in the presence of 15.4, 23.1 and 34.6 microM zinc. Such lowered activities of ADA were restored to normal when the cells were cultured in the presence of higher zinc concentration equal to 53.8 and 73.1 microM. From the results we concluded that it is possible to restore ADA activity in aged skin fibroblasts to normal levels by raising the zinc concentration in the culture medium to four or five times the control normal plasma zinc level. PMID- 12910481 TI - A combination of alpha-tocopherol, vitamin C and N-acetyl cysteine increases unsaturated fatty acid levels in hydrogen peroxide-induced Candida tropicalis (ATCC 13803). AB - This research was aimed at evaluating the antioxidant effects of combinations of alpha lipoic acid (LA), vitamin C (VC), N-acetyl cysteine (NAC) and alpha tocopherol (TOC) on lipid level and fatty acid composition of C. tropicalis (ATCC 13803) against hydrogen peroxide toxicity. According to the experimental results, the cell density of C. tropicalis increased significantly in NAC+LA+H2O2, NAC+TOC+ H2O2 and NAC+VC+H2O2 groups (p<0.001) at the end of 48 and 72 h incubation times. The total lipid level in H2O2 and H2O2 + antioxidant supplemented groups was lower than that of the control group. In the fatty acid composition of C. tropicalis, the palmitic acid level was raised in the NAC group (p<0.05), whereas its level was reduced in the other supplemented groups. While the oleic acid level increased in NAC+TOC+H2O2 and NAC+VC+H2O2 (p<0.001) groups, its level slightly decreased in the H2O2 group. The linolenic acid level was low in all the supplemented groups, but linoleic acid and total mono-unsaturated fatty acid (MUFA) levels were high in these groups compared with the control group. Total polyunsaturated fatty acid level (PUFA) decreased in NAC and H2O2 groups (p<0.01), but its level increased in NAC+LA+H2O2 and NAC+TOC+H2O2 groups (respectively, p<0.01, p<0.001). Total saturated fatty acid level decreased significantly in NAC+TOC+H2O2, NAC+H2O2 and NAC+VC+H2O2 (p<0.001) groups (p<0.01), whereas total unsaturated fatty acid level increased in NAC, NAC+H2O2, NAC+LA+H2O2, NAC+TOC+H2O2 and NAC+VC+H2O2 groups. In conclusion, our data showed that the levels of total unsaturated fatty acid, MUFA and PUFA were raised with the combinations of NAC and TOC, LA and VC in C. tropicalis cells subjected to hydrogen peroxide toxicity. PMID- 12910483 TI - Effects of caffeic acid phenethyl ester and alpha-tocopherol on reperfusion injury in rat brain. AB - Oxygen-derived free radicals have been implicated in the pathogenesis of cerebral injury after ischaemia-reperfusion. Caffeic acid phenethyl ester (CAPE), an active component of propolis extract, exhibits antioxidant properties. The purpose of the present study was to investigate the effects of ischaemia and subsequent reperfusion on rat brain and to investigate the effects of two free radical scavengers, CAPE and alpha-tocopherol, on this in vivo model of cerebral injury. Ischaemia was induced by bilateral occlusion of the carotid arteries for 20 min and reperfusion was achieved by releasing the occlusion to restore the circulation for 20 min. Control rats underwent a sham operation. CAPE at 10 micromol kg(-1) or alpha-tocopherol at 25 micromol kg(-1) was administered intraperitoneally before reperfusion. Reperfusion led to significant increase in the activity of xanthine oxidase and higher malondialdehyde levels in the brain. Acute administration of both CAPE and alpha-tocopherol suppressed ischaemia reperfusion-induced cerebral lipid peroxidation and injury, but CAPE seems to offer a better therapeutic advantage over alpha-tocopherol. PMID- 12910485 TI - Do multivitamin or folic acid supplements reduce the risk for congenital heart defects? Evidence and gaps. AB - Congenital heart defects are among the most common congenital anomalies and are the leading cause of infant death due to congenital anomalies. Except for a few known measures, effective primary prevention is not yet feasible for most heart anomalies. Recent reports have associated the use of multivitamin supplements around the time of conception and during early pregnancy with a reduced risk for heart defects in the offspring. We review and discuss the evidence and suggest a framework for further investigation in this area. PMID- 12910484 TI - Antioxidant status and lipid peroxidation in type II diabetes mellitus. AB - Diabetes Mellitus (DM), a state of chronic hyperglycaemia, is a common disease affecting over 124 million individuals worldwide. In this study, erythrocyte glutathione levels, lipid peroxidation, superoxide dismutase, catalase, and glutathione peroxidase and some extracellular antioxidant protein levels of patients with type II diabetes mellitus and healthy controls were investigated. Thirty-eight patients (21 males; with age of mean +/- SD, 53.1+/-9.7 years) and 18 clinically healthy subjects (10 males; with age of mean +/- SD, 49.3+/-15.2 years) were included in the study. Levels of erythrocyte lipid peroxidation, serum ceruloplasmin and glucose levels, HbA1C levels, and erythrocyte catalase activity were significantly increased, whereas serum albumin and transferrin levels, erythrocyte glutathione levels, and glutathione peroxidase activity were significantly decreased compared to those of controls. There was no significant difference in superoxide dismutase activity compared to controls. The results suggest that the antioxidant deficiency and excessive peroxide-mediated damage may appear in non-insulin dependent diabetes mellitus. PMID- 12910486 TI - Use of a multiplex PCR/sequencing strategy to detect both connexin 30 (GJB6) 342 kb deletion and connexin 26 (GJB2) mutations in cases of childhood deafness. AB - Hearing loss is a common congenital disorder that is frequently associated with mutations in the Cx26 gene (GJB2). Three recent reports that found a large deletion in another DFNB1 gene, Cx30 (GJB6), suggest that this defect may cause nonsyndromic recessive hearing loss through either a homozygous deletion of Cx30, or digenic inheritance of a Cx30 deletion and a Cx26 mutation in trans. We designed a simple diagnostic strategy with multiplex PCR followed by direct sequencing to allow for the simultaneous detection of Cx26 mutations and Cx30 deletions, and evaluated its effectiveness as a clinical genetic test by examining 200 DNA samples. In the 108 samples from deaf subjects, two digenic mutations were identified in Cx26 and Cx30 (E47X/342 kb deletion and 167delT/342 kb deletion); 69 had only Cx26 mutations (29 biallelic, 40 singleton), including two novel frameshift mutations 511-512insAACG and 358-360delAG; and 37 had no detectable mutation in either Cx26 or Cx30. Our deletion mapping suggested that the proximal breakpoint of all reported Cx30 large deletions are between the nucleotide 444 and 627 at the Cx30 coding region within a maximal interval of 78 or 184 bp. This simultaneous examination of Cx26 and Cx30 is a practical and efficient diagnostic approach for patients with nonsyndromic congenital deafness. PMID- 12910487 TI - MCA/MR syndrome with hypocholesterolemia related to familial dominant hypobetalipoproteinemia. AB - Recent reports have emphasized the role of cholesterol in vertebrate embryonic development. The RSH or so-called Smith-Lemli-Opitz syndrome (SLOS) was the first multiple congenital anomalies/mental retardation syndrome related to a cholesterol synthesis disturbance. Familial hypobetalipoproteinemia is a well known dominantly inherited entity in which affected individuals usually are free of symptoms. We report on the unusual association of a malformation syndrome with mental retardation resembling SLOS and profound hypocholesterolemia related to familial hypobetalipoproteinemia. We discuss the possible causal relationship between the two conditions and the current understanding of the role of cholesterol in normal embryogenesis. PMID- 12910488 TI - Congenital deficiency of alpha-fetoprotein and associated chromosomal abnormality in the placenta. AB - In this study we describe two patients with congenital absence of alpha fetoprotein (AFP). The pathological examination results, including an immunohistochemical stain, which define qualitatively the levels of AFP detected by the biochemical studies and the comparative genomic hybridization (CGH) are enclosed. A description of the suggested functions of AFP and the means of its production are set forth. An explanation is suggested for the lack of symptoms in a newborn with undetectable levels of AFP and the mechanism by which this condition might occur. PMID- 12910489 TI - Talo-patello-scaphoid osteolysis, synovitis, and short fourth metacarpals in sisters: a new syndrome? AB - Osteolysis syndromes are characterized by resorption of affected bones with associated swelling and pain. Various forms of multicentric osteolysis syndromes including autosomal dominant and recessive carpal-tarsal osteolysis, Torg, Francois, Whyte-Hemingway, Hajdu-Cheney, Winchester, and other forms have been described. Most present in pre-school years with extensive involvement and destruction of multiple bones. We present a sister-pair, both of whom presented in early teenage, i.e., 13 and 15.5 years, respectively, with bilateral ankle, knee, and later, wrist pain. Radiological examination revealed bilateral osteolysis of tali, scaphoids, and patellae, and short fourth metacarpals in both sisters. Further investigation revealed absence of renal involvement, a normal excretion of amino acids, mucopolysaccharides and oligosaccharides, and presence of chronic synovitis in both sisters. Both parents and a younger brother were without radiographic or clinical evidence of the disease and there was no history of consanguinity. Thus, our sister-pair presented with the same carpal and tarsal bone involvement at a much later age, with evidence of chronic synovitis, along with short fourth metacarpals (brachydactyly type E changes) and without renal disease, suggesting a new syndrome with probable autosomal recessive inheritance. PMID- 12910490 TI - Clinical and diagnostic characteristics of complex III deficiency due to mutations in the BCS1L gene. AB - We investigated two siblings of a Spanish family presenting with congenital lactic acidosis. They had severe failure to thrive, liver dysfunction, and renal tubulopathy. An isolated biochemical complex III deficiency was detected in liver. A search for mutations in the human bc1 synthesis like (BCS1L) gene was undertaken. Direct sequencing revealed a missense mutation R45C and a nonsense mutation R56X, both located in exon 1 of BCS1L. The missense mutation in combination with a loss of function of the second allele is responsible for the isolated complex III deficiency in this family. PMID- 12910491 TI - Segmental neurofibromatosis in childhood. AB - Segmental neurofibromatosis refers to individuals who have manifestations of neurofibromatosis type 1 (NF-1) limited to one area of the body. It results from a post-conceptional mutation in the NF-1 gene leading to somatic mosaicism. Although it is generally considered a rare condition, this report of 39 children with segmental NF-1 demonstrates that it is commonly seen in a pediatric NF-1 referral center. The mean age at diagnosis was 7.8 years (range: 2-25 years). Twenty-nine patients had only pigmentary manifestations of segmental NF-1, including seven who had only cafe-au-lait macules and 22 who had cafe-au-lait macules and freckling. Two patients had isolated plexiform neurofibromas; a third patient had a plexiform neurofibroma of the eyelid in addition to ipsilateral dysplasia of the sphenoid wing and Lisch nodules. A 12-year-old girl had an isolated tibial pseudarthrosis. An 8-year-old boy had an isolated optic pathway tumor, which behaved both biologically and radiographically as an NF1-associated tumor. While most children with segmental NF-1 have only localized pigmentary changes, some children will have isolated plexiform neurofibromas, pseudarthroses, or optic pathway tumors. Accurate diagnosis of segmental NF-1 is crucial for both management and genetic counseling. PMID- 12910493 TI - Molecular characterization of an inherited ring (19) demonstrating ring opening. AB - Ring chromosomes arise following breakage in both chromosome arms and rejoining of the centric segment at the broken ends or by end-to-end fusion of the telomeres. The phenotype of ring carriers is unpredictable, and developmental abnormalities may occur even when the ring appears to be structurally balanced. This is believed to be due to mitotic instability from abnormal segregation and sister chromatid exchange in somatic cells. Although ring chromosomes usually arise as de novo events, transmittal from mosaic carriers to offspring sometimes occurs. In such cases, offspring with ring mosaicism in combination with a normal cell line remain unexplained. In this report, we used detailed molecular and cytogenetic analyses of a prenatally detected, inherited ring (19) to observe the behavior of the ring chromosome in culture, and to investigate the mechanism of inherited ring chromosome mosaicism. PMID- 12910492 TI - Growth hormone receptor variant (L526I) modifies plasma HDL cholesterol phenotype in familial hypercholesterolemia: intra-familial association study in an eight generation hyperlipidemic kindred. AB - Defect of growth hormone receptor (GHR) is classically known to cause Laron syndrome, characterized by short stature, specific facial appearance, elevated serum growth hormone levels, and decreased insulin-like growth factor I levels. In addition, an increased cardiovascular risk due to elevated plasma total and LDL cholesterol levels marks another feature of the disease. Growth hormone (GH) plays an important role in the regulation of lipoprotein metabolism. GH status was found to be an independent determinant of plasma total cholesterol and triglyceride levels in humans. We studied a total of 207 members of eight generation extended family of familial hypercholesterolemia (FH) in which affected members presented with various lipoprotein phenotypes. Intra-familial correlation analysis of a modifier effect of a Leu526Ile substitution in GHR gene was carried out among 95 carriers for LDL receptor gene (LDLR) mutation and 112 non-carriers. When plasma high-density lipoprotein cholesterol (HDL-c) levels in the LDLR-mutation carriers were compared, a significant lowering effect of HDL-c was observed with the Leu allele; the values were lowest among Leu/Leu homozygotes (mean +/- SD = 37 +/- 2 mg/dl), highest in Ile/Ile homozygotes (50 +/ 4 mg/dl), and intermediate among Leu/Ile heterozygotes (41 +/- 2 mg/dl) (P = 0.0021). The results indicate a significant modification of the phenotype of FH with the defective LDLR allele, by GHR Leu variation in the kindred studied. PMID- 12910494 TI - ABase-a tool for the rapid assessment of anthropometric measurements on handheld computers. AB - Anthropometry is widely recognized as an important method in the evaluation of dysmorphic children. Nevertheless, it has never gained widespread clinical use. We believe that this has mainly practical reasons: appropriate growth charts are not readily available at the bedside or in clinic and taking multiple measurements and plotting them into corresponding growth charts is a time consuming task. Here we describe a computer program that overcomes both problems: ABase compares entered anthropometric measurements to a database of age- and sex matched reference values, calculates the centile rank and displays the result either as text or as a digitized growth chart. The program runs on small handheld computers that can easily be carried around in a shirt pocket. PMID- 12910495 TI - Overlap between VACTERL and hemifacial microsomia illustrating a spectrum of malformations seen in axial mesodermal dysplasia complex (AMDC). AB - We describe a male infant born to consanguineous healthy parents with multiple congenital anomalies of the skeleton and internal organs. His phenotype displays an overlap between VACTERL and hemifacial microsomia (oculo-auriculo-vertebral spectrum/OAV). In addition, striking asymmetry of the malformations further supports the classification as part of the "axial mesodermal dysplasia complex" (AMDC) which is supposed to arise from disturbed mesodermal cell migration during early blastogenesis. The present case was instructive to review the continuous spectrum of AMDC related anomalies. PMID- 12910498 TI - Characterization of a supernumerary ring chromosome 1 mosaicism in two cell systems by molecular cytogenetic techniques and review of the literature. AB - We report on a 4-year-old boy with developmental delay and microcephaly with an additional small marker chromosome derived from chromosome 1 and detected in 14% of T-lymphocytes by conventional cytogenetics and in 9% of buccal smear cells by interphase FISH. Using molecular cytogenetic techniques, the marker chromosome was characterized as an extra ring chromosome consisting of euchromatic material from the proximal short arm of chromosome 1. We compare the cytogenetic data and the phenotype of our patient to those previously described cases with marker chromosome 1 mosaicism. We conclude that in addition to the straightforward molecular cytogenetic characterization of the euchromatic content of the ring chromosome, the investigation of a second cell system gives additional information about the tissue specific distribution of the supernumerary marker chromosome (SMC) and provides more reliable data for further karyotype/phenotype correlations and the prediction of the phenotypic outcome in prenatal cases. PMID- 12910497 TI - Prostate cancer is part of the hereditary non-polyposis colorectal cancer (HNPCC) tumor spectrum. AB - The recognized urologic tumor spectrum in hereditary non-polyposis colon cancer includes ureteral and renal pelvis malignancies. Here, we report a family in which the proband, who had three metachronous adenocarcinomas of the colon and rectum (at ages 54, 57, and 60), presented with an adenocarcinoma of the prostate at age 61. Immunohistochemical (IHC) staining of colonic, rectal, and prostatic tumor tissues demonstrated lack of expression of both MSH2 and MSH6. Accordingly, microsatellite instability (MSI) was found in the rectal, colonic, and prostatic tumors. The kindred complies with the Amsterdam criteria for HNPCC, as five members over three generations had colorectal cancer. Molecular investigations were initiated when the proband's son presented with an adenocarcinoma of the colon at age 35. Southern blotting analysis of genomic DNA led to identification of a novel genomic deletion encompassing exon 5 of the MSH2 gene. Although prostate cancer has occasionally been described in HNPCC families, to the best of our knowledge, this is the first report where the MSI and IHC analysis of the prostatic adenomcarcinoma clearly link its aetiology to the germline mismatch repair mutation. Hence, prostate cancer should be included in the HNPCC tumor spectrum. PMID- 12910496 TI - Cardiovascular abnormalities associated with the Stuve-Wiedemann syndrome. AB - The Stuve-Wiedemann syndrome (SWS) is a congenital bone dysplasia characterized by camptodactyly with ulnar deviation and congenital bowing of the long bones. Affected patients present with respiratory difficulties in the neonatal period or later and recurrent episodes of hyperthermia. The typical radiological findings are bowing of the long bones of the lower limbs, wide metaphyses with decreased density, and abnormal trabecular pattern. Generally, respiratory insufficiency and hyperthermia are reported to be the cause of death. We report on two sibs with SWS, who died from severe pulmonary hypertension with pulmonary artery wall abnormality. We suggest a common pathophysiological process, which could explain the cardiovascular findings that we observed immediately after birth in the two affected sibs. We hypothesize that the severe pulmonary hypertension due to the arterial wall abnormality could explain the neonatal death of these two children. PMID- 12910499 TI - Isolated postaxial polydactyly type B with mosaicism of a submicroscopic unbalanced translocation leading to an extended phenotype in offspring. AB - Postaxial polydactyly (PAP) is characterized by the presence of one or more extra ulnar or fibular digits or parts of it. PAP type B presents frequently as a skin tag on the hand(s). It is usually an isolated malformation, but in 6.6% it is associated with other congenital abnormalities, mostly well recognizable syndromes. We present a male with PAP-B only and his daughter with an extended phenotype including mental retardation and minor dysmorphisms. Both share a cytogenetically balanced t(4;7)(p15.2;q35), present in mosaicism in the father. We found microdeletions associated with the breakpoints. The chromosomal regions described here have not been previously associated with the PAP-B phenotype. We present the first case of an individual with isolated PAP-B and a submicroscopic chromosome abnormality. PMID- 12910500 TI - Premature thelarche in Coffin-Siris syndrome. PMID- 12910501 TI - Primary pulmonary dysgenesis in velocardiofacial syndrome: a second patient. PMID- 12910502 TI - Under-ascertainment of mosaic carriers of balanced homologous acrocentric translocations and isochromosomes. AB - Acrocentric rearrangements are the most common chromosome abnormalities in humans. Carriers of homologous acrocentric rearrangements (Robertsonian translocations (ROBs) between homologous chromosomes and isochromosomes) are at very high risk of having multiple spontaneous abortions and chromosomally abnormal offspring. Parents of fetuses and children with unbalanced homologous acrocentric rearrangements are rarely found to be carriers or mosaic for the same rearrangement. Even though recurrent miscarriages may indicate a carrier parent, carriers are rarely identified. Comparison of non-chromosome 21 homologous rearrangements to rea(21q21q) culled from the literature revealed a 7-fold decrease in the number of mosaic cases among the parents of non-rea(21q21q) offspring. This under-ascertainment in parents may be due to low level mosaicism confined to the gonads, a true biological difference between chromosome 21 rearrangements and other homologous acrocentric rearrangements, or simply to the lack of rigorous clinical investigation of the parental karyotypes to uncover mosaicism. We recommend that polymorphic marker analysis be applied to apparently de novo acrocentric rearrangements to distinguish those resulting from biparental postzygotic formation from those resulting from meiotic formation; the latter of which may indicate a potential carrier parent. Parental chromosomal constitutions could then be screened in a large number of cells and in more than one tissue type to identify mosaicism. Identification of mosaicism allows for accurate genetic counseling and discussion of reproductive options. However, given that mosaicism may be restricted to the gonads, prenatal testing is likely to be desired by the family whether or not mosaicism is found. PMID- 12910503 TI - Clinical and molecular diagnosis should be consistent. PMID- 12910504 TI - Comments on "Epidemiological evidence that maternal diabetes does not appear to increase the risk for Down syndrome". PMID- 12910507 TI - Abstracts of the XXXV meeting of the International Society of Paediatric Oncology (SIOP) and the International Society of Paediatric Surgical Oncology (IPSO). Cairo, Egypt, October 8-11, 2003. PMID- 12910508 TI - Hepatocellular carcinoma: is there a potential for chemoprevention using cyclooxygenase-2 inhibitors? AB - Inhibitors of cyclooxygenase-2 (COX-2) have proapoptotic and antiangiogenic effects on malignant tumors and inhibit their invasion to surrounding tissues. These properties are derived from COX-dependent and/or COX-independent signaling via peroxisome proliferator-activated receptor gamma. Although the role of COX-2 involvement in human hepatocarcinogenesis has not been determined yet, selective COX-2 inhibitors with COX-independent properties may potentially suppress hepatocarcinogenesis. This hypothesis should be confirmed in in vivo studies using animal models. These studies may provide insights into any application of the COX-2 inhibitor for primary and/or secondary chemoprevention. PMID- 12910509 TI - Where do we stand with hepatoblastoma? A review. AB - Hepatoblastoma (HB) is the most common pediatric liver malignancy, comprising approximately 1% of all pediatric cancers. The disparate clinical staging systems and histologic classifications that were developed during the last decades, nevertheless, reflect the remaining difficulties and uncertainties in characterizing HB. Furthermore, the combination of surgery and (neo)adjuvant chemotherapy has improved patient outcomes dramatically. A poor prognosis is associated with large tumor size, multifocality, extrahepatic disease, and metastatic spread. The exact etiology of HB remains unknown, but the cytogenetic alterations, phenotypic features, and biologic aspects that accompany this neoplasm yield more and more insight into its pathogenesis. New cell-biologic and molecular-biologic insights may lead to the development of new treatment modalities, especially for patients with a bad prognosis. This review summarizes the different aspects of this intriguing tumor and discusses the current status of research and treatment for patients with HB. PMID- 12910510 TI - Long-term adjustment of survivors of early-stage breast carcinoma, 20 years after adjuvant chemotherapy. AB - BACKGROUND: The long-term impact of breast carcinoma and its treatment was assessed in 153 breast carcinoma survivors previously treated on a Phase III randomized trial (Cancer and Leukemia Group B [CALGB 7581]) a median of 20 years after entry to CALGB 7581. METHODS: Survivors were interviewed by telephone using the following standardized measures: Brief Symptom Inventory (BSI), PostTraumatic Stress Disorder Checklist with the trauma defined as survivors' response to having had cancer (PCL-C), Conditioned Nausea, Vomiting and Distress, European Organization for Research and Treatment of Cancer QLQ-C30 (quality of life), Life Experience Survey (stressful events), MOS Social Support Survey, comorbid conditions (Older Americans Resources and Services Questionnaire), and items developed to assess long-term breast carcinoma treatment side effects and their interference with functioning. RESULTS: Only 5% of survivors had scores that were suggestive of clinical levels of distress (BSI), 15% reported 2 or more posttraumatic stress disorder (PTSD) symptoms (PCL-C) that were moderately to extremely bothersome, 1-6% reported conditioned nausea, emesis, and distress as a consequence of sights, smells, and tastes triggered by reminders of their treatment, 29% reported sexual problems attributed to having had cancer, 39% reported lymphedema, and 33%, reported numbness. Survivors who reported greater lymphedema and numbness that interfered with functioning had significantly worse PTSD (PCL-C; P = 0.008) com- pared with survivors who reported less lymphedema and numbness. Survivors with a lower level of education (P = 0.026), less adequate social support (P = 0.0033), more severe negative life events (P = 0.0098), and greater dissatisfaction with their medical care (P = 0.037) had worse PTSD compared with other survivors. CONCLUSIONS: Twenty years after the initial treatment, the impact of breast carcinoma on survivors' adjustment was minimal. However, the higher prevalence of PTSD symptoms in response to having had cancer is indicative of continuing psychologic sequelae long after treatment completion. Findings related to lymphedema and numbness and continued symptoms of PTSD suggest that the long-term psychologic and medical sequelae on adjustment may be underrecognized. To establish in more detail whether survivors' overall psychologic state is any different from that of individuals without cancer, a population of community residents without cancer would need to be studied. PMID- 12910511 TI - Short-term morbidity of the upper limb after sentinel lymph node biopsy or axillary lymph node dissection for Stage I or II breast carcinoma. AB - BACKGROUND: The goals of sentinel lymph node biopsy (SLNB) are to improve axillary staging and reduce unnecessary axillary lymph node dissections (ALND), thereby reducing treatment-related upper-limb morbidity. In the current prospective study, short-term upper-limb morbidity was assessed after SLNB and/or ALND. METHODS: The study comprised 204 patients with Stage I/II breast carcinoma. Mean patient age was 55.6 years (standard deviation, 11.6). Sixty-six patients (32%) underwent SLNB only, and 138 (68%) underwent a Level I-II ALND. Assessment (preoperative [t0] and 6 weeks postoperative [t1]) included evaluation of shoulder range of motion, muscle strength, grip strength, pain, upper/forearm circumference, shoulder disability, and activities of daily life (ADL). RESULTS: Considerable treatment-related upper-limb morbidity was observed. Significant (P < 0.001) changes were found for pain, range of motion in forward flexion, abduction and abduction/external rotation, strength of shoulder abductors and elbow flexors, and in perceived disability in ADL. However, no significant difference in change of upper-limb function and ADL was found between the SLNB and ALND groups. CONCLUSIONS: Significant short-term treatment-related upper-limb morbidity exists after SLNB or ALND. There is no significant difference in short term treatment-related morbidity between SLNB and ALND. PMID- 12910512 TI - Eighteen-year results in the treatment of early breast carcinoma with mastectomy versus breast conservation therapy: the National Cancer Institute Randomized Trial. AB - BACKGROUND: Between 1979-1987, the National Cancer Institute conducted a randomized, prospective study of mastectomy (MT) versus breast conservation therapy (BCT) in the treatment of patients with early-stage breast carcinoma. After a median potential follow-up of 18.4 years, the authors present the updated results. METHODS: After informed consent was obtained from each patient, 237 evaluable women with clinical AJCC Stage I and Stage II breast carcinoma were enrolled on an institutionally reviewed protocol and randomly assigned to undergo modified radical MT (116 patients) or BCT (121 patients), which was comprised of lumpectomy, axillary lymph node dissection, and radiation therapy. Negative surgical margins in the lumpectomy arm were not required. The 237 randomized patients were followed for a median potential follow-up of 18.4 years. The primary endpoints were overall survival and disease-free survival. RESULTS: At a median follow-up of 18.4 years, there was no detectable difference with regard to overall survival between patients treated with MT and those treated with BCT (58% vs. 54%; P = 0.67 overall). Twenty-seven women in the BCT arm (22%) experienced an in-breast event. After censoring in-breast events in the BCT arm that were salvaged successfully by MT, disease-free survival also was found to be statistically similar (67% in the MT arm vs. 63% in the BCT arm; P = 0.64 overall). There was no statistically significant difference with regard to contralateral breast carcinoma between the two treatment arms (P = 0.70). CONCLUSIONS: After nearly 20 years of follow-up, there was no detectable difference in overall survival or disease-free survival in patients with early stage breast carcinoma who were treated with MT compared with those treated with BCT. For BCT patients, long-term in-breast failures continued to occur throughout the duration of follow-up. There was no statistically significant difference in the incidence of contralateral breast carcinoma between the two treatment groups. PMID- 12910514 TI - Enhancement of lymph node metastasis and distant metastasis of thyroid carcinoma. AB - BACKGROUND: The mechanisms of local and distant metastases are imperfectly understood. The goal of the current study was to add to the body of knowledge regarding local and distant metastases of thyroid malignancies. METHODS: The authors performed multivariate analysis of 573 patients who underwent surgery between November 1994 and May 2002 for follicular (FTC; n = 100), papillary (PTC; n = 236), or medullary thyroid carcinoma (MTC; n = 237) at a university hospital. RESULTS: In multivariate analysis, extrathyroidal extension consistently evolved as the key risk factor for both lymph node metastasis and distant metastasis. This correlation was most pronounced in MTC and least pronounced in FTC. The risk of lymph node metastasis also increased with reoperative status in patients with MTC and with primary tumor diameter in patients with MTC (tumor diameter > 10 mm) and patients with PTC (tumor diameter > 20 mm). In the PTC group, lymph node metastasis was more common among patients younger than age 45. In the MTC group, extrathyroidal growth and distant metastasis were associated exclusively with lymph node metastasis. Lymph node metastasis was the only secondary risk factor for distant metastasis. In the analysis of risk factors for distant metastasis in the FTC and PTC groups, no interaction was found between extrathyroidal growth and lymph node metastasis. This finding suggests that extrathyroidal growth and lymph node metastasis of FTC and PTC, and presumably also MTC, represent separate mechanisms and routes of distant metastasis. CONCLUSIONS: Screening for both local residual disease and distant metastases should be intensified in the high risk population of patients whose primary tumors exhibit large diameters, extrathyroidal growth, or lymph node metastasis. PMID- 12910513 TI - Androgen receptors frequently are expressed in breast carcinomas: potential relevance to new therapeutic strategies. AB - BACKGROUND: Several studies have demonstrated the biologic and therapeutic significance of estrogen and progesterone receptors (ER and PR) in breast carcinomas. The aim of the current study was to examine the presence of androgen receptors (AR) in breast carcinomas. METHODS: Two hundred cases of breast carcinoma, consisting of 145 invasive and 55 noninvasive (ductal carcinoma in situ [DCIS]) lesions, were examined using a monoclonal antibody against AR on formalin-fixed, paraffin-embedded archival material. The results were analyzed for correlations with immunohistochemically determined ER, PR, and HER-2/neu expression. RESULTS: Eighty-seven of the 145 cases (60%) of invasive carcinoma and 45 of the 55 cases (82%) of DCIS were AR-positive according to internationally standardized guidelines. The vast majority of Grade 1 carcinomas were positive for AR (90% of invasive Grade 1 carcinomas and 95% of Grade 1 DCIS), whereas in Grade 3 invasive carcinomas and DCIS, positive immunoreactions for AR were observed in 46% and 76% of cases, respectively. Among the cases of Grade 3 carcinoma, 33 invasive carcinomas (39%) and 17 DCIS lesions (68%) were ER negative but AR-positive. Among Grade 1 carcinomas (invasive and DCIS), not a single case was positive for HER-2/neu, but most cases were intensely positive for AR. In contrast, many invasive Grade 3 carcinomas exhibited agreement between AR status and HER-2/neu status (AR-positive and HER-2/neu-positive, 30.5%; AR negative and HER-2/neu-negative, 42.5%). CONCLUSIONS: Androgen receptors are commonly expressed in DCIS and in invasive breast carcinoma. A significant number of poorly differentiated carcinomas are ER-negative and PR-negative but AR positive. Immunohistochemical examination of AR would be desirable because it would provide additional information about steroid receptors in breast carcinomas. PMID- 12910515 TI - Expression and regulation of the novel vascular endothelial growth factor receptor neuropilin-1 by epidermal growth factor in human pancreatic carcinoma. AB - BACKGROUND: It was recently shown that neuropilin-1 (NRP-1), which was described originally as a receptor for the semaphorins/collapsins (ligands involved in neuronal guidance), is a coreceptor for vascular endothelial growth factor (VEGF) and increases the affinity of specific isoforms of VEGF to its receptor, VEGF-R2. METHODS: The authors investigated the expression and regulation of NRP-1 in human pancreatic adenocarcinoma specimens and cell lines. RESULTS: Immunohistochemical analysis revealed that NRP-1 was expressed in 12 of 12 human pancreatic adenocarcinoma specimens but was absent in nonmalignant pancreatic tissue. Northern blot analysis revealed NRP-1 mRNA expression in 8 of 11 human pancreatic adenocarcinoma cell lines. NRP-1 mRNA expression was increased by epidermal growth factor (EGF) but not by tumor necrosis factor alpha in several of the human pancreatic adenocarcinoma cell lines studied. Treating human Panc-48 adenocarcinoma cells with EGF activated Akt and Erk but not P-38. Blockade of the phosphatidylinositol-3 kinase (PI-3K)/Akt, mitogen-activated protein kinase (MAPK)/Erk, or P-38 pathways abrogated EGF-induced NRP-1 expression. Finally, EGF receptor blockade in vivo led to a decrease in NRP-1 expression in an orthotopic model of human pancreatic carcinoma. CONCLUSIONS: NRP-1 is expressed in most human pancreatic adenocarcinomas and cell lines but not in nonmalignant pancreatic tissue. EGF regulates NRP-1 expression through the PI-3K/Akt and MAPK/Erk signaling pathways, and blockade of the EGF receptor is associated with decreased expression of NRP-1 in vivo. NRP-1 may act as a coreceptor for VEGF in pancreatic carcinoma, as it does in other tumor systems, thereby enhancing angiogenesis and the effect of VEGF on the growth of pancreatic adenocarcinoma. PMID- 12910516 TI - The significance of thymidine phosphorylase/platelet-derived endothelial cell growth factor activity in renal cell carcinoma. AB - BACKGROUND: Thymidine phosphorylase (TP) is identical to platelet-derived endothelial cell growth factor (PDECGF) and has angiogenic activity. Little is known about the significance of TP/PDECGF activity in patients with renal cell carcinoma (RCC). The authors examined the enzymatic activity of TP in 66 RCC specimens and investigated the association between the level of TP activity and the stage/grade status of patients with RCC. Furthermore, the authors examined the correlation between TP/PDECGF activity and prognosis. METHODS: TP activity levels in nonfixed, fresh-frozen RCC specimens and in specimens of normal kidney were determined using a thin-layer chromatography assay. RESULTS: The activity of TP was approximately 3.5-fold greater in RCC specimens compared with normal kidney specimens. TP activity in patients with Stage III-IV RCC was 2.6-fold greater compared with TP activity in patients with Stage I-II RCC. In addition, the level of TP activity was correlated with a higher grade of RCC. Patients who had RCC with low TP activity had a longer postoperative disease-specific survival compared with patients who had RCC with high TP activity in the 5-year follow-up. CONCLUSIONS: The current study is the first to demonstrate a correlation between levels of TP activity and both disease progression and a higher grade of RCC. It also is the first to show that elevated TP activity in patients with RCC predicts a poor prognosis. The results suggest that high TP/PDECGF activity may be associated with the malignant potential of RCC and that TP/PDECGF may be a molecular therapeutic target in patients with RCC. PMID- 12910518 TI - Advanced seminoma--treatment results and prognostic factors for survival after first-line, cisplatin-based chemotherapy and for patients with recurrent disease: a single-institution experience in 145 patients. AB - BACKGROUND: Advanced seminoma is a rare clinicopathologic entity. To the authors' knowledge, very few sizeable reports published to date have studied the outcome of patients with advanced seminoma after first-line and salvage therapy, and few have dealt with prognostic factors initially or in patients with recurrent disease. METHODS: The records of 145 men with advanced seminoma who were treated with cisplatin-based first-line chemotherapy regimens were reviewed. Six patient characteristics, including age, prior radiotherapy, primary tumor site, initial serum lactate dehydrogenase and human chorionic gonadotropin levels, and disease stage, were studied as initial prognostic factors. In patients with recurrent disease, outcome according to the site of recurrence and the salvage treatment was also reviewed. RESULTS: A complete response was obtained in 130 patients (90%) after cisplatin-based first-line chemotherapy, and the 5-year overall survival rate was 81% (95% confidence interval [95% CI], 73-87%). Nonpulmonary visceral metastasis at diagnosis was the only initial adverse prognostic factor. Thirty-one patients (21%) developed recurrent disease. Recurrence in the liver or the central nervous system was a major adverse prognostic factor, with a 5-year overall survival rate of 7% (95% CI, 1-32%), compared with 58% (95% CI, 33-79%) in patients who had lymph node, lung, or bone recurrences. The only durable complete remission after a liver recurrence was obtained with high-dose chemotherapy followed by autologous stem cell transplantation. All 12 patients who were treated for primary mediastinal seminoma with cisplatin-based chemotherapy alone were long-term disease free survivors. CONCLUSIONS: Overall, the prognosis of patients with advanced seminoma was good after cisplatin-based, first-line chemotherapy. Metastasis in the liver or the central nervous system, initially or at recurrence, is currently the only proven adverse prognostic factor. PMID- 12910517 TI - Identification of fibroblast growth factor receptor 3 mutations in urine sediment DNA samples complements cytology in bladder tumor detection. AB - BACKGROUND: Mutations in fibroblast growth factor 3 receptor (FGFR3) are frequent events in low-grade bladder tumors. To assess the potential utility of the detection of FGFR3 mutations in a screening modality, the authors analyzed urine sediment DNA samples from 192 patients in a retrospective study. METHODS: Urine sediment DNA samples from 192 patients were prepared. Seventy-two patients had undergone transurethral resection (TURBT group) of mainly Ta lesions and 120 patients had undergone cystectomy (cystectomy group). The majority of patients in the cystectomy group had more advanced tumors compared with patients in the TURBT group. DNA preparations were screened for FGFR3 mutations in exons 7, 10, and 15 using single-strand conformation polymorphism (SSCP) and DNA sequencing. RESULTS: Using SSCP, 67% of patients in the TURBT group and 28% in the cystectomy group displayed FGFR3 mutations. Comparative analysis of cytology results and FGFR3 mutational analysis were performed in 122 cases. Within the TURBT group, FGFR3 mutation analysis outperformed cytology. FGFR3 mutation analysis identified change in 68% of urine sediment DNA samples whereas cytology recorded the presence of tumor cells in 32% of the DNA samples. In the cystectomy group, cytology outperformed FGFR3 mutation analysis. Cytology recorded tumor detection in 90% of patients, while SSCP identified mutational change in 24%. CONCLUSIONS: Combining FGFR3 mutation results with cytology in both groups correctly identified tumor presence in 105 of 122 (86%) of patients. The greater sensitivity of FGFR3 mutation detection over cytology in identifying the presence of low-grade, superficial bladder tumors represents a potential new tool to complement standard cytology in screening patients for bladder tumors and recurrent disease. PMID- 12910519 TI - Does retroperitoneal lymph node dissection have a curative role for patients with sex cord-stromal testicular tumors? AB - BACKGROUND: Sex cord-stromal tumors account for < 5% of all adult testicular tumors, and 10% are malignant. Due to the limited reported experience, there is no agreement on the best management, especially in patients who have tumors with malignant pathologic features or who present with metastatic disease. The authors attempt to evaluate the role of retroperitoneal lymph node dissection (RPLND) in the management of patients with these malignant sex cord-stromal tumors. METHODS: Reviewing the Indiana University testis cancer registry revealed 17 patients who underwent RPLND for sex cord-stromal tumors. Pathology was reviewed for features suggestive of malignancy. The data examined included clinical and pathologic stage, surgical procedure, additional therapy received, and outcome. RESULTS: Pathology included Leydig tumors in six patients, Sertoli tumors in four patients, sex cord-stromal tumors in five patients, a granulosa cell tumor in one patient, and a poorly differentiated non-germ cell tumor in one patient. Nine patients had histologic features suggestive of malignancy. Clinical stage at surgery was Stage I in nine patients and Stage IIA-IIIA in eight patients. Patients underwent modified or bilateral RPLND. Nine patients had pathologic Stage I tumors, and the remaining eight patients and had pathologic Stage IIB IIIA tumors. Follow-up ranged from 8 months to 11 years. Of the eight patients with Stage II-III disease, six patients eventually died of metastatic disease despite additional radiotherapy and/or chemotherapy. CONCLUSIONS: Sex cord stromal tumors have a potentially aggressive malignant behavior that is difficult to predict based on clinical and pathologic features. Although the therapeutic role of RPLND in patients with small-volume metastatic retroperitoneal tumors is unclear, RPLND remains an option to be performed immediately after orchiectomy, especially in patients who have tumors with malignant features and/or small volume metastatic disease. PMID- 12910520 TI - Expression of c-ABL, c-KIT, and platelet-derived growth factor receptor-beta in ovarian serous carcinoma and normal ovarian surface epithelium. AB - BACKGROUND: Tyrosine kinases, such as c-KIT, c-ABL, and platelet-derived growth factor-beta (PDGFR-beta), are important regulators of cell growth. Highly potent and selective inhibitors of tyrosine kinases are being investigated as alternatives to standard chemotherapy. One such inhibitor, imatinib mesylate, is being used to treat gastrointestinal stromal tumors and chronic myelogenous leukemia. Ovarian carcinomas frequently develop resistance to conventional chemotherapeutic agents. Immunohistochemical expression of c-ABL, PDGFR-beta, and c-KIT was evaluated in ovarian carcinomas to determine whether treatment with imatinib mesylate might be feasible. METHODS: The expression of c-ABL, c-KIT, and PDGFR-beta in tumors was evaluated by immunohistochemical analysis of 52 ovarian serous carcinomas, including 21 low-grade (well differentiated) and 31 high-grade (poorly differentiated) tumors. Fourteen normal ovaries were also evaluated. RESULTS: In normal ovarian surface epithelium, c-ABL was expressed universally. PDGFR-beta was expressed in the majority (93%) of samples of normal ovarian epithelium, whereas the c-KIT protein was undetectable in normal ovarian surface epithelium. Overall, c-ABL was expressed in 71% of serous carcinomas. c-ABL was expressed more frequently in the low-grade serous carcinomas (81%) compared with the high-grade serous carcinomas (65%). PDGFR-beta expression was observed in 81% of serous carcinomas overall and was observed more frequently in higher-grade tumors. c-KIT immunohistochemical staining was absent in low-grade tumors but was present in 26% of high-grade serous carcinomas. CONCLUSIONS: The majority of ovarian serous carcinomas express one or more of the kinases targeted by the tyrosine kinase inhibitor, imatinib mesylate, suggesting the potential usefulness of this drug in the treatment of ovarian carcinoma. PMID- 12910521 TI - Radiotherapy for patients with early-stage glottic carcinoma: univariate and multivariate analyses in a group of consecutive, unselected patients. AB - BACKGROUND: Radiotherapy (RT) has a remarkable success rate in the treatment of patients with glottic carcinoma. The objectives of the current study were to assess the results in a group of consecutive patients with comparable characteristics who were treated with RT (6-megavolt photon linear accelerator) and to determine the prognostic factors that may influence local control in patients with early-stage glottic carcinoma. The impact on local control of tobacco smoking and second primary malignancies also was investigated. METHODS: Four hundred ten patients with T1-T2 squamous cell carcinoma of the glottis who were treated between 1986 and 2001 were analyzed retrospectively with regard to local control and overall survival. Potential prognostic factors for local control were evaluated with univariate and multivariate models. The impact of technologic advances also was evaluated. RESULTS: The 5-year and 10-year overall survival rates were 83% and 63.5%, respectively. The overall 10-year local control rate for patients with T1-T2 glottic carcinoma was 89%. The median time to recurrence was 7 months. Univariate analysis showed that tumor category, tumor size, macroscopic appearance of the lesion, RT fraction size, persistent edema, year of RT treatment, unchanged dysphonia, and surgical option all had a significant influence on local control; whereas multivariate analysis showed that only persistent dysphonia and year of RT treatment were significantly associated with increased local control. A 22.2% rate of second primary malignancies was reported: second primary tumors were the major cause of death in the patients studied. Only 2 patients died of laryngeal carcinoma; 304 patients were alive with their disease in complete remission, 1 patient was alive with recurrent laryngeal carcinoma after undergoing salvage surgery, and 103 patients died of either intercurrent disease or a second primary tumor. CONCLUSIONS: The use of a 6-megavolt photon linear accelerator achieved a high rate of local control in patients with T1-T2 glottic carcinoma. Dysphonia and the year of RT treatment were the most important prognostically significant factors for patient outcome. The occurrence of a second primary tumor was the most frequent cause of death, especially among patients who did not stop smoking after a diagnosis of glottic carcinoma. PMID- 12910522 TI - Phase II study of alemtuzumab in chronic lymphoproliferative disorders. AB - BACKGROUND: Pilot studies showed that alemtuzumab is active in lymphoproliferative disorders. The authors conducted a Phase II trial to evaluate the efficacy and safety of alemtuzumab in advanced or refractory chronic lymphoproliferative disorders. METHODS: Seventy-eight patients were enrolled. The most common diagnoses were chronic lymphocytic leukemia (n = 42 patients) and T cell prolymphocytic leukemia (n = 18 patients). Before entering the study, the patients had received multiple therapies (median, three therapies) and the median time from diagnosis was 7 years. Alemtuzumab was given intravenously at doses of 3 mg, 10 mg, and 30 mg on 3 consecutive days, after which 30 mg was administered 3 times a week. Patients were treated for 4-12 weeks depending on disease response. All patients received prophylactic trimethoprim/sulfamethoxazole and valacyclovir. RESULTS: The overall response rate was 35%, the complete response (CR) rate was 13%, and the partial response (PR) rate was 22%. The median duration of response was 18 months for patients achieving a CR and 7 months for patients achieving a PR. The median duration of survival was 25 months for patients who had a response and 12 months for the entire population. Normalization of the lymphocyte count was observed in 84% of patients and resolution of bone marrow involvement was observed in 49% of patients. The most common infusion-related adverse events were fever, rigors, skin rash, nausea, and dyspnea. These were most common during the first week of therapy. Hematologic toxicity was comprised of long-lasting lymphocytopenia and transient neutropenia and thrombocytopenia. Thirty-six patients (46%) experienced at least one episode of fever or infection. CONCLUSIONS: Alemtuzumab has a high response rate in patients with chronic lymphoproliferative disorders. PMID- 12910523 TI - The impact of age on toxicity, response rate, quality of life, and survival in patients with advanced, Stage IIIB or IV nonsmall cell lung carcinoma treated with carboplatin and paclitaxel. AB - BACKGROUND: The optimal treatment strategy for elderly patients with advanced nonsmall cell lung carcinoma has not been defined to date. The authors performed a retrospective analysis of a Phase III trial that treated patients who had Stage IIIB or IV nonsmall cell lung carcinoma with carboplatin and paclitaxel and analyzed the impact of age on response rate, survival, toxicity, and quality of life. METHODS: Patients with Stage IIIB or IV NSCLC were randomized to receive either 4 cycles of carboplatin at an area under the curve (AUC) of 6 and paclitaxel at a dose of 200 mg/m(2) every 21 days or treatment with carboplatin and paclitaxel (C/P) until they developed disease progression. At the time of disease progression, all patients on both arms were to receive second-line weekly paclitaxel at a dose of 80 mg/m(2) per week. In this analysis, patients age 70 years and older were compared with patients younger than age 70 years. In addition, a minimum log rank P value analysis was performed in an attempt to identify other potential age splits that may have been significant. RESULTS: Two hundred thirty patients were randomized. Sixty-seven patients were age 70 years or older (29%). The median number of cycles delivered for both age groups was 4 cycles (range, 0-19 cycles). No statistically significant differences in any of the most common toxicities (Grade >or= 2) associated with C/P were identified (data from Cycles 1-4) for patients younger than age 70 years compared with patients age 70 years and older, respectively, including neutropenia (38% vs. 35%), neuropathy (13% vs. 16%), leukopenia (7% vs. 13%), myalgia/arthralgia (15% vs. 9%), malaise (8% vs. 15%), anemia (9% vs. 4%), thrombocytopenia (7% vs. 9%), anorexia (8% vs. 4%), and nausea/emesis (14% vs. 15%). In addition, no potential age splits that may have been significant were found using a minimum log rank P value analysis. CONCLUSIONS: The current analysis demonstrated that C/P exhibited similar toxicity profiles in patients age 70 years and older compared with patients younger than age 70 years. The survival rates were not different between the two age groups, and there was no difference in progression of quality-of-life outcomes. In fit, elderly patients, C/P represented a reasonable standard regimen. PMID- 12910524 TI - Vascular endothelial growth factor-C expression correlates with lymph node localization of human melanoma metastases. AB - BACKGROUND: Melanoma metastasizes by different mechanisms comprising direct invasion of the surrounding tissue and spreading via the lymphatic or vascular system. Despite their clinical relevance, the molecular mechanisms that guide the route of spreading and localization of the metastases in different tissues are not well known. Recent studies in different tumor types have shown that vascular endothelial growth factor-C (VEGF-C), which displays a high specificity for lymphatic endothelium, is involved in tumor-induced lymphangiogenesis and lymphatic metastatic spread. The authors studied the expression of VEGF-C in cultured human melanoma cells derived from cutaneous and lymph node metastases as well as in metastatic melanoma tissue specimens to assess a possible involvement of this growth factor in lymph node localization of melanoma metastases. METHODS: VEGF-C expression was evaluated in vitro on human melanoma cell lines established from cutaneous and lymph node metastasis specimens by reverse transcriptase polymerase chain reaction, Northern blot analysis, and immunofluorescence analysis. Immunohistochemical analysis of 42 tissue specimens of melanoma metastases and 10 tissue specimens of primary skin melanomas was also performed. RESULTS: Preferential expression of VEGF-C was detected in lymph node-derived tumor cell lines at both the mRNA and protein levels. The association between VEGF-C production and lymph node localization of metastases was confirmed by the in vivo analysis. In addition, analysis of 10 patients, from whom specimens of both the primary skin melanoma and melanoma metastases were available, indicated a correlation between VEGF-C expression in the primary tumor and lymph node localization of metastases. CONCLUSIONS: The findings of the current study demonstrate that VEGF-C expression is correlated with localization of melanoma metastases in the lymph nodes and suggest that VEGF-C expression in primary skin melanoma may be predictive of lymph node metastatic dissemination. PMID- 12910525 TI - Characterization of the neoplastic phenotype in the familial atypical multiple mole melanoma-pancreatic carcinoma syndrome. AB - BACKGROUND: Previous studies suggest that the familial atypical multiple-mole melanoma (FAMMM) syndrome may predispose affected families to nonmelanoma carcinomas, including adenocarcinoma of the pancreas. It has been found that many of these families harbor mutations in the CDKN2A gene on chromosome 9p21. The phenotypic expression of CDKN2A mutations in these families has not been characterized fully. METHODS: The authors studied eight families that appeared to inherit multiple nevi, cutaneous melanomas, and pancreatic carcinomas in association with a CDKN2A germline mutation. The expression of disease within these families was examined, and segregation ratios were estimated to assess the patterns of inheritance according to various definitions of phenotype. RESULTS: Either multiple nevi or pancreatic carcinoma was diagnosed in 53% of first-degree relatives of the probands. The offspring of parents affected with multiple nevi, melanoma, or pancreatic carcinoma were significantly more likely to be affected themselves compared with the offspring of unaffected parents (48.9% vs. 16.7%; P = 0.004). CONCLUSIONS: The current results provide additional evidence that multiple nevi, melanoma, or pancreatic carcinoma may be inherited as autosomal dominant traits in families known to harbor CDKN2A mutations. Other malignancies may be a part of the phenotype in these families, although this hypothesis requires additional study. PMID- 12910526 TI - Myoid differentiation and prognosis in adult pleomorphic sarcomas of the extremity: an analysis of 92 cases. AB - BACKGROUND: The results of a recent study demonstrated an association between myoid differentiation and an adverse prognosis in adult patients with pleomorphic sarcoma, as determined by 5-year metastasis-free survival rates. METHODS: To confirm the importance of muscle differentiation on prognosis in a well controlled clinical context, 92 samples from patients with pleomorphic sarcoma of the extremity from a single institution were immunostained with 4 monoclonal antibodies believed to be correlated with myoid differentiation: alpha-smooth muscle actin, muscle-specific actin, desmin, and myoglobin. RESULTS: Forty-two cases were positive for at least 1 muscle marker and 50 cases were uniformly negative. Between the two groups, there was no significant difference in tumor size, tumor extent, or patient age found; however, histologic grade was significantly higher (P = 0.038) in the myoid tumors. The 5-year survival differed significantly between patients with myoid tumors (35%) and those without myoid tumors (65%) (P = 0.0054). Myoid differentiation remained an adverse prognostic indicator after adjusting for clinically significant factors (i.e., histologic grade, tumor size, tumor extent, and patient age) (P = 0.01) (hazard ratio, 2.39; 95% confidence interval, 1.24-4.63). Furthermore, there was an inverse relation found between the number of myoid markers present and survival (P = 0.004). CONCLUSIONS: Myoid differentiation was found to be an independent indicator of adverse prognosis in adult patients with pleomorphic spindle cell sarcoma of the extremity. PMID- 12910527 TI - Obesity as a potential risk factor for adenocarcinomas and squamous cell carcinomas of the uterine cervix. AB - BACKGROUND: Hormonal factors may play a more prominent role in cervical adenocarcinoma than squamous cell carcinoma. The authors evaluated whether obesity, which can influence hormone levels, was associated with adenocarcinoma and squamous cell carcinoma. METHODS: This case-control study included 124 patients with adenocarcinoma, 139 matched patients with squamous cell carcinoma, and 307 matched community control participants. All participants completed interviews and provided cervicovaginal samples for human papillomavirus (HPV) testing. Polytomous logistic regression-generated odds ratios (ORs) and 95% confidence intervals (95% CIs) for self-reported height and weight, body mass index (BMI; kg/m(2)), and measured waist-to-hip ratio (WHR) for both histologic types were adjusted and stratified for HPV and other confounders. RESULTS: Height, weight, BMI, and WHR were positively associated with adenocarcinoma. BMI >or= 30 kg/m(2) (vs. BMI < 25 kg/m(2); OR, 2.1 and 95% CI, 1.1-3.8) and WHR in the highest tertile (vs. the lowest tertile; OR, 1.8 and 95% CI, 0.97-3.3) were associated with adenocarcinoma. Neither height nor weight was found to be associated with squamous cell carcinoma, and associations for BMI >or= 30 kg/m(2) (OR, 1.6) and WHR in the highest tertile (OR, 1.6) were weaker and were not statistically significant. Analyses using only HPV positive controls showed similar associations. The data were adjusted for and stratified by screening, but higher BMI and WHR were associated with higher disease stage at diagnosis, even among recently and frequently screened patients with adenocarcinoma. Thus, residual confounding by screening could not be excluded as an explanation for the associations. CONCLUSIONS: Obesity and body fat distribution were associated more strongly with adenocarcinoma than with squamous cell carcinoma. Although questions about screening remain, obesity may have a particular influence on the risk of glandular cervical carcinoma. PMID- 12910528 TI - Measuring the side effects of taxane therapy in oncology: the functional assesment of cancer therapy-taxane (FACT-taxane). AB - BACKGROUND: Cancer chemotherapy with some of the taxane class of agents can be associated with significant neurotoxicity, arthralgias, myalgias, and skin changes that may offset the therapeutic benefits of taxane use. METHODS: The authors developed and tested a set of questions to assess these important side effects of taxane therapy from the patient's perspective. The current study evaluated the taxane subscale of the Functional Assessment of Chronic Illness Therapy (FACIT) measurement system. Reliability, validity, and responsiveness to expected change were evaluated in the context of an ongoing clinical trial comparing four cycles of carboplatin plus paclitaxel with a strategy of carboplatin plus paclitaxel until disease progression in patients with advanced nonsmall cell lung carcinoma (NSCLC). RESULTS: The 16-item Taxane subscale score and the 11-item peripheral neuropathy subset both demonstrated excellent internal consistency and concurrent validity, and the scores worsened as one would predict during a 12-week treatment course of taxane therapy. Results of the psychometric analyses supported the use of this subscale for measuring the unwanted adverse consequences of effective cancer therapies. Measuring the patient perception of treatment side effects also allowed a preliminary exploration of the relative quality of life (QOL) impact of symptom relief and treatment toxicity. The results indicated that toxicity and symptom improvement may make relatively equivalent contributions to total QOL as measured by the summary score from a multidimensional QOL instrument, the Functional Assessment of Cancer Therapy General. However, symptom status and improvement appear to play a stronger role than taxane toxicity in patients' global rating of their QOL. CONCLUSIONS: Future research might examine this question of competing benefits as a potential aid to decision-making regarding the administration of toxic therapies in the setting of advanced disease. PMID- 12910530 TI - Identification of genes that are regulated transcriptionally by Myc in childhood tumors. AB - BACKGROUND: Amplification of the N-myc oncogene is associated with adverse outcomes in the common childhood tumor, neuroblastoma. Because the transforming properties of Myc are related to its ability to modulate gene expression, the authors used cDNA microarrays to identify potential Myc target genes. METHODS: Expression levels of 4608 genes were analyzed in a series of neuroblastoma cell lines. Identical analyses were performed in a panel of medulloblastoma cell lines to identify c-Myc targets and to determine the extent to which N-Myc targets and c-Myc targets were shared. Comparisons were made between cell lines with high levels versus low levels of Myc protein expression. RESULTS: Array analyses yielded 121 genes with increased expression levels (>or= 1.65-fold) and 9 genes with decreased expression levels in N-Myc-expressing versus nonexpressing cell lines. Many of these were newly identified targets of biologic interest. Fifty percent of the N-Myc targets (60 of 121) were mutual c-Myc targets. A significant correlation between the level of N-myc and selected target gene expression was demonstrated independently in 27 neuroblastoma tumor samples and in an N-myc inducible cell line system. CONCLUSIONS: A number of diverse pathways are modulated by N-Myc in neuroblastoma. Although, overall, there was significant correlation between myc and target transcript expression among cohorts of tumors, great variability in levels of target expression was seen among individual tumor samples, and this biologic heterogeneity in the levels of target gene expression may offer insight into differences in the clinical behavior of neuroblastoma and may prove to be of prognostic significance in the future. PMID- 12910529 TI - Phase II study of ecteinascidin 743 in heavily pretreated patients with recurrent osteosarcoma. AB - BACKGROUND: Recurrent osteosarcoma is a drug-resistant disease with a dismal prognosis. The objective of this Phase II study was to evaluate the activity of ecteinascidin 743 (ET-743) as a salvage therapy in these patients. METHODS: Patients with recurrent osteosarcoma who had received standard chemotherapeutic agents were eligible. ET-743 was administered at a dose of 1500 microg/m(2) as a 24-hour infusion every 3 weeks. Pharmacokinetic studies were performed during the first cycle. RESULTS: Twenty-five patients were enrolled, 23 of whom were assessable for response (median age of 18 years; range, 12-67 years). The median number of previous chemotherapeutic agents was five (range, three to eight previous agents). Sixty-one cycles were administered (median number of cycles per patient was 2; range, 1-9 cycles per patient). Three patients (12%) achieved minor responses (49% 36% and 25%, respectively). Fifteen patients (60%) developed a transient elevation of hepatic transaminases (Grade 3 or 4 [according to the National Cancer Institute Common Toxicity Criteria]), which was not cumulative. Grade 3 or 4 neutropenia and thrombocytopenia were observed in 12 patients (48%) and 6 patients (24%), respectively. The mean area under the curve (AUC) in 4 patients experiencing Grade 4 toxicity (76.4 +/- 29.3 ng x hr/mL) was significantly greater (P = 0.034) than that in those for whom the most severe toxicity was Grade 3 (39.5 +/- 17.2 ng x hr/mL [n = 12]) or Grade 1-2 (52.6 +/- 15.6 ng x hr/mL [n = 5]). There were no other significant correlations found between pharmacokinetic variables and patient characteristics, toxicity, or therapeutic response. CONCLUSIONS: ET-743 was found to be well tolerated in heavily pretreated osteosarcoma patients but had limited antitumor activity as a single agent. The combination of ET-743 with cisplatin or doxorubicin should be considered. PMID- 12910531 TI - Cognitive-behavioral intervention for distress in patients with melanoma: comparison with standard medical care and impact on quality of life. AB - BACKGROUND: Melanoma accounts for > 79% of skin cancer-related deaths, although it accounts for only 4% of skin cancer incidence. Given the potential for lethality, it is likely that patients with melanoma may experience significant emotional distress. The current study was designed to determine the effect of a cognitive-behavioral intervention on distress and health-related quality of life (HRQOL) in patients with melanoma who had medium-to-high distress. METHODS: Forty eight patients who had Global Severity Index scores >or= 60 2 months after their initial visit to the multidisciplinary melanoma clinic were randomized to receive either standard care or 4 sessions of a cognitive-behavioral intervention (CBI). Repeated assessments using the Brief Symptom Inventory, the Medical Outcomes Survey Short Form-36, and the State-Trait Anxiety Inventory occurred at baseline, at 2 months, and at 6 months after intervention for both groups. RESULTS: An intent-to-treat analysis did not reveal significantly lower distress in the CBI group at 2 months or 6 months of follow-up, although differences were noted in anxiety and HRQOL. An effect-of-intervention analysis did reveal lower levels of distress in the CBI group at 2 months, with differences approaching significance at 6 months. CONCLUSIONS: The four-session CBI significantly reduced distress and improved HRQOL for a period of 2 months in patients with melanoma who had medium to-high distress, with improved general health evident 6 months after the intervention. Some variation in results was revealed in an intent-to-treat analysis. The initial evidence from the current study showed that a brief intervention may be effective for creating change in individuals with cancer who have increased distress, although further research is needed to identify the most optimal approach for delivering the intervention. PMID- 12910532 TI - Radiotherapy for extranodal, marginal zone, B-cell lymphoma of mucosa-associated lymphoid tissue originating in the ocular adnexa: a multiinstitutional, retrospective review of 50 patients. AB - BACKGROUND: Due to the small number of patients and differences in the pathologic classification in most radiotherapy series, information regarding the adequacy of tumor control in patients with ocular-adnexal marginal zone B-cell lymphoma of mucosa-associated lymphoid tissue (MALT lymphoma) is limited. METHODS: A multiinstitutional, retrospective study was performed on 50 patients with Stage IE ocular-adnexal MALT lymphoma who were treated with radiotherapy between 1989 and 1999. The impact of patient characteristics and other variables on tumor control was analyzed. RESULTS: Responses to radiotherapy include a complete response (CR) in 26 patients, a partial response (PR) in 20 patients, and no change in 4 patients. Forty-nine of 50 patients obtained tumor control in the ocular adnexa at 24 months. Overall, 6 patients exhibited disease recurrence at 4 97 months. Three patients developed recurrence in the ocular adnexa. Two patients had isolated extranodal failure involving the oral floor and the submandibular gland, and one patient experienced failure in the neck lymph node. The initial tumor response had a marginal impact on the development of recurrence. None of the 26 patients who achieved a CR experienced ocular-adnexal recurrence. All three patients who experienced local treatment failure belonged to the initial PR group. In total, five of six patients who developed recurrent disease had obtained a PR after initial radiotherapy. Age, gender, tumor location, and dose of radiotherapy did not influence the development of recurrence. There was only one death due to lymphoma. The 5-year overall survival rate was 91% with a median follow-up of 46 months. CONCLUSIONS: Radiotherapy offers excellent local control with a prolonged clinical course for patients with MALT lymphoma in the ocular adnexa. The initial response to radiotherapy marginally influenced the probability of recurrence. PMID- 12910534 TI - Cancer chronotherapy: principles, applications, and perspectives. PMID- 12910533 TI - Detection of UGT1A10 polymorphisms and their association with orolaryngeal carcinoma risk. AB - BACKGROUND: UGT1A10 exhibits glucuronidating activity against metabolites of the tobacco smoke carcinogen, benzo(a)pyrene, and is expressed highly in numerous target tissues for tobacco-related cancers including the upper aerodigestive tract. The current study was conducted to determine the prevalence of genetic polymorphisms in the UGT1A10-specific region of the UDP-glucuronosyltransferase family 1A locus and their relationship with risk for orolaryngeal carcinoma. METHODS: The authors analyzed UGT1A10-specific sequences in a population of black, white, and Asian individuals. Ten UGT1A10 alleles were identified by direct sequencing of UGT1A10 sequences amplified by polymerase chain reaction (PCR) using DNA purified from buccal cell swabs that were taken from individual subjects. RESULTS: In addition to three silent polymorphisms, three missense polymorphisms were found at codons 139 (Glu > Lys), 240 (Thr > Met), and 244 (Leu > Ile). Using PCR-restriction fragment length polymorphism analysis of buccal cell DNA, the prevalence of the UGT1A10(240Met) variant was less than 0.01% in whites and blacks. Similarly, the prevalence of both the UGT1A10(139Lys) and UGT1A10(244Ile) variants was less than 0.01% in whites but it was significantly higher (0.04 and 0.05, respectively, P < 0.01) in blacks. None of the missense UGT1A10 variants were found in any of the Asian individuals examined. In a case control study of black individuals, a significant association with orolaryngeal carcinoma risk was found in persons with at least 1 UGT1A10(139Lys) allele (crude odds ratio, 0.29 [95% confidence interval, 0.10-0.81]; adjusted odds ratio, 0.20 [95% confidence interval, 0.05-0.87]). No association was observed for the codon 244 (Leu > Ile) polymorphism. CONCLUSIONS: The data from the current study show that the UGT1A10 gene has several low-frequency missense polymorphisms and that the codon 139 polymorphism is an independent risk factor for orolaryngeal carcinoma in blacks. PMID- 12910536 TI - Silencing of retroviral vector transduced LacZ reporter gene by frameshift mutation. AB - Moloney murine leukemia virus-based vector expressing Escherichia coli beta galactosidase (lacZ) as reporter gene and the transposon Tn5 neomycin resistance (neo) gene was transduced at low-multiplicity of infections into NIH 3T3 cells. Geneticin (G418)-resistant cells were recloned and cell lines containing beta galactosidase positive or beta-galactosidase negative cells were obtained. Both positive and negative cell lines contained a single proviral copy at distinct integration sites. RNA complementary to lacZ was detected in beta-galactosidase positive as well as in one of three investigated beta-galactosidase negative cell lines. DNA sequence analysis of proviral LacZ gene in beta-galactosidase negative cell line C6 showed a single nucleotide insertion at position 1567 resulting in reading frame shift and translational stop codon at position 1629. This mutation explains the enzyme inactivation. The absence of beta-galactosidase after retroviral transduction of LacZ reproter gene may be a consequence of definite mutation but not a consequence of ineffective transduction or transcriptional inactivation of transgene. PMID- 12910537 TI - Purification of bioethanol effluent in an UASB reactor system with simultaneous biogas formation. AB - In this study, the prospect of using an Upflow Anaerobic Sludge Blanket (UASB) reactor for detoxification of process water derived from bioethanol production has been investigated. The bioethanol effluent (BEE) originated from wet oxidized wheat straw fermented by Saccharomyces cerevisiae and Thermoanaerobacter mathranii A3M4 to produce ethanol from glucose and xylose, respectively. In batch experiments the methane potential of BEE was determined to 529 mL-CH(4)/g-VS. In batch degradation experiments it was shown that the presence of BEE had a positive influence on the removal of the inhibitors 2-furoic acid, 4 hydroxyacetophenone, and acetovanillone as compared to conversion of the inhibitors as sole substrate in synthetic media. Furthermore, experiments were carried out treating BEE in a laboratory-scale UASB reactor. The results showed a Chemical Oxygen Demand (COD) removal of 80% (w/w) at an organic loading rate of 29 g-COD/(L. d). GC analysis of the lignocellulosic related potentially inhibitory compounds 2-furoic acid, vanillic acid, homovanillic acid, acetovanillone, syringic acid, acetosyringone, syringol, 4-hydroxybenzoic acid, and 4-hydroxybenzaldehyde showed that all of these compounds were removed from the BEE in the reactor. Implementation of a UASB purification step was found to be a promising approach to detoxify process water from bioethanol production allowing for recirculation of the process water and reduced production costs. PMID- 12910538 TI - In-situ near infrared spectroscopy to monitor key analytes in mammalian cell cultivation. AB - The use of in-situ near infrared spectroscopy (NIRS) as a tool for monitoring four key analytes in a CHO-K1 animal cell culture was investigated. Previous work using on-line NIRS to monitor bioprocesses has involved its application ex-situ where the analyzer is physically outside the fermentor, or to microbial bioprocesses. This novel application of NIRS to monitor analytes within an animal cell culture using a steam sterilizable in-situ fiber optic probe is very important for furthering the use of NIRS within the bioprocessing industry. The method of calibration used to develop the models involved the use of large data sets so that all likely variation in stoichiometry was incorporated within the models. Successful models for glucose, lactate, glutamine, and ammonia were built with Standard Error of Predictions (SEP's) of 0.072 (g/L), 0.0144 (g/L), 0.308 (mM), and 0.036 (mM), respectively of the total concentration range. PMID- 12910539 TI - Modeling ionic hydrogels swelling: characterization of the non-steady state. AB - Ionic hydrogels can be used as controlled release systems that respond to an external substrate or trigger by swelling or de-swelling. One example is a glucose-sensitive system for insulin-controlled release based on pH-sensitive hydrogel. To enhance understanding of non-steady state swelling, and to facilitate design of specifications (e.g., glucose-sensitivity) of the pH sensitive ionic hydrogel based on the copolymer poly (2-hydroxyethyl methacrylate co-N, N-dimethylaminoethyl methacrylate) (poly (HEMA-co-DMAEMA)), we developed a mathematical compartmental model using the software SAAM II. Current analytical and computational methods focus on equilibrium swelling of hydrogels; although for many stimuli-responsive hydrogel applications, the dynamic process is significant. We now report, using a combination of experimental data and kinetic analysis that in the poly (HEMA-co-DMAEMA) the rate of proton entry is governed by a different rate coefficient than water entry rate. The transport coefficient governing water uptake is dependent upon three variables: pH of external media, amine groups incorporated into the polymer, and crosslinking density of the polymer. An additional result is that swelling equilibrium is reached when all the amine groups are protonated. In this study we also demonstrate the predictive capability of the model for both interpolated and extrapolated data, and its use in design of future bench experiments. Uncovering these fundamental properties of pH-sensitive hydrogels with the aid of a kinetic model suggests that the complexities of hydrogel research and development can be overcome by combining experimental and computational approaches. PMID- 12910540 TI - Numerical simulation and PEPT measurements of a 3D conical helical-blade mixer: a high potential solids mixer for solid-state fermentation. AB - Helical-blade solids mixers have a large potential as bioreactors for solid-state fermentation (SSF). Fundamental knowledge of the flow and mixing behavior is required for robust operation of these mixers. In this study predictions of a discrete particle model were compared to experiments with colored wheat grain particles and positron emission particle tracking (PEPT) measurements. In the discrete particle model individual movements of particles were calculated from interaction forces. It was concluded that the predicted overall flow behavior matched well with the PEPT measurements. Differences between the model predictions and the experiments with wheat grains were found to be due to the assumption that substrate particles were spherical, which was in the model. Model simulations and experiments with spherical green peas confirmed this. The mixing in the helical-blade mixer could be attributed to (1) the transport of particles up and down in the interior of the mixer, and (2) dispersion or micro-mixing of particles in the top region of the mixer. It appeared that the mixing rate scaled linearly with the rotation rate of the blade, although the average particle velocity did not scale proportionally. It may be that the flow behavior changes as a function of the rotation rate (e.g., changing thickness of the top region); further study is required to confirm this. To increase the mixing performance of the mixer, a larger blade or a change in the shape of the mixer (larger top surface/volume ratio) is recommended. PMID- 12910541 TI - Long-term continuous evolution of acetate resistant Acetobacter aceti. AB - Elevated concentrations of cytotoxic acetate are found in many environmental niches, and few species are relatively resistant to acetate. In particular the high-level acetate resistance of so-called acetic acid bacteria that occurs in industrial settings must be constantly selected for. To investigate the nature of such high-level resistance, we grew the moderately acetate-resistant Acetobacter aceti wild-type and acetate-sensitive Escherichia coli in long-term continuous cultures with increasing acetate concentrations at near neutral pH. While E. coli did not acquire any significant resistance after 125 generations of selection, A. aceti evolved the capability to grow at acetate concentrations exceeding 50 g/L within 240 generations. This phenotype was found to be stable for several generations in the absence of selective pressure, hence must be genetically determined. Intracellular acetate concentrations were significantly lower in evolved A. aceti, when compared to wild-type A. aceti and E. coli, indicating that cytoplasmatic anion accumulation is an important component of acetate toxicity. PMID- 12910542 TI - Quantification of central metabolic fluxes in the facultative methylotroph methylobacterium extorquens AM1 using 13C-label tracing and mass spectrometry. AB - The metabolic fluxes of central carbon metabolism were measured in chemostat grown cultures of Methylobacterium extorquens AM1 with methanol as the sole organic carbon and energy source and growth-limiting substrate. Label tracing experiments were carried out using 70% (13)C-methanol in the feed, and the steady state mass isotopomer distributions of amino acids derived from total cell protein were measured by gas chromatography coupled to mass spectrometry. Fluxes were calculated from the isotopomer distribution data using an isotopomer balance model and evolutionary error minimization algorithm. The combination of labeled methanol with unlabeled CO(2), which enters central metabolism in two different reactions, provided the discriminatory power necessary to allow quantification of the unknown fluxes within a reasonably small confidence interval. In wild-type M. extorquens AM1, no measurable flux was detected through pyruvate dehydrogenase or malic enzyme, and very little flux through alpha-ketoglutarate dehydrogenase (1.4% of total carbon). In contrast, the alpha-ketoglutarate dehydrogenase flux was 25.5% of total carbon in the regulatory mutant strain phaR, while the pyruvate dehydrogenase and malic enzyme fluxes remained insignificant. The success of this technique with growth on C(1) compounds suggests that it can be applied to help characterize the effects of other regulatory mutations, and serve as a diagnostic tool in the metabolic engineering of methylotrophic bacteria. PMID- 12910543 TI - Site-specific recombination in human cells catalyzed by the wild-type integrase protein of coliphage HK022. AB - The activity of the Integrase (Int) protein encoded by coliphage HK022 was tested in a human cell culture. Plasmids were constructed as substrates that carry the sites of the integration reaction (attP and attB) or the sites of excision (attL and attR). The site-specific recombination reactions were monitored in cis and in trans configurations by the expression of the green fluorescent protein (GFP) as a reporter. Cells cotransfected with the substrate plasmid(s) and with a plasmid that expresses the wild-type Int show efficient integration as well as excision in both configurations. The wild-type Int was active in the human cells without the need to supply the accessory proteins integration host factor (IHF) and excisionase (Xis) that are indispensable for the reaction in the bacterial host. PMID- 12910544 TI - Chemical processing as a tool to generate ovalbumin variants with changed stability. AB - Processing of ovalbumin may result in proteins that differ more than 23 degrees C in denaturation temperature while the structural fold is not significantly affected. This is achieved by 1) conversion of positive residues into negative ones (succinylation); 2) elimination of negative charges (methylation); 3) reducing the proteins hydrophobic exposure (glycosylation); 4) increasing the hydrophobic exposure (lipophilization); or by 5) processing under alkaline conditions and elevated temperature (S-ovalbumin). The effect on the structural fold was investigated using a variety of biochemical and spectroscopic tools. The consequences of the modification on the thermodynamics of the protein was studied using differential scanning calorimetry and by monitoring the tryptophan fluorescence or ellipticity at 222 nm of protein samples dissolved in different concentrations of guanidine-HCl. The impact of the modification on the denaturation temperature scales for all types of modifications with a free energy change of about 1 kJ per mol ovalbumin per Kelvin (or 0.0026 kJ per mol residue per K). The nature of the covalently coupled moiety determines the impact of the modification on the protein thermodynamics. It is suggested that especially for lipophilized protein the water-binding properties are substantially lowered. Processing of globular proteins in a controlled manner offers great opportunities to control a desired functionality, for example, as texturizer in food or medical applications. PMID- 12910545 TI - Use of confocal scanning laser microscopy to measure the concentrations of aerial and penetrative hyphae during growth of Rhizopus oligosporus on a solid surface. AB - In order to develop a method for use in investigations of spatial biomass distribution in solid-state fermentation systems, confocal scanning laser microscopy was used to determine the concentrations of aerial and penetrative biomass against height and depth above and below the substrate surface, during growth of Rhizopus oligosporus on potato dextrose agar. Penetrative hyphae had penetrated to a depth of 0.445 cm by 64 h and showed rhizoid morphology, in which the maximum biomass concentration, of 4.45 mg dry wt cm(-3), occurred at a depth of 0.075 cm. For aerial biomass the maximum density of 39.54 mg dry wt (-3) occurred at the substrate surface. For both aerial and penetrative biomass, there were two distinct regions in which the biomass concentration decayed exponentially with distance from the surface. For aerial biomass, the first exponential decay region was up to 0.1 cm height. The second region above the height of 0.1 cm corresponded to that in which sporangiophores dominated. This work lays the foundation for deeper studies into what controls the growth of fungal hyphae above and below the surfaces of solid substrates. PMID- 12910546 TI - Transient self-inhibition of the growth of Lactobacillus delbrueckii subsp. bulgaricus in a pH-regulated fermentor. AB - An industrial strain of Lactobacillus delbrueckii subsp. bulgaricus was grown in a synthetic medium on lactose as carbon substrate, in a pH-regulated fermentor. Growth proceeded in two distinct phases separated by a transient stationary phase. Various experimental approaches were used to identify the cause of this growth arrest. Growth experiments in L. bulgaricus culture supernatant fluids collected at different cultivation times in fermentor, and supplemented or not with various nutritional solutions, enabled us to discard the possibility of a nutritional limitation. Tube cultures of L. bulgaricus in medium supplemented with various lactic acid concentrations showed a potential inhibition by this metabolic end product but confirmed that this inhibition was not responsible for the cessation of growth. It was concluded that at least one inhibitory compound was produced during the growth phase of the strain, and this compound disappeared from the medium in the transient stationary phase, enabling the growth to start again later in the culture. Indeed, the stoichiometric analysis of the culture showed, firstly, that unidentified carbon compounds were produced from lactose during growth, which were probably converted in lactic acid during the transient stationary phase and, secondly, that part of the amino acids consumed gave catabolic end products. Finally, bacteriocin-like compounds were not considered to be responsible for this growth arrest. PMID- 12910547 TI - Simplified modeling of fed-batch alcoholic fermentation of sugarcane blackstrap molasses. AB - Simplified modeling based on material balances for biomass, ethanol and substrate was used to describe the kinetics of fed-batch alcohol fermentation of sugarcane blackstrap molasses. Maintenance requirements were previously shown to be of particular significance in this system, owing to the use of massive inoculum to minimize inhibitions; therefore, they were taken into consideration for kinetic modeling. Average values of biomass and ethanol yields, productivities, and substrate consumption rates, calculated at the end of runs performed either at constant or exponentially varying flow rates, demonstrated that all of these parameters were influenced by the initial sugar-feeding rate, F(o)S(o). Under conditions of substrate shortage (F(o)S(o) 17 years old) who used oral NSAIDs for more than 4 days. Two reviewers evaluated 4,881 published titles, identifying 55 NSAID versus placebo randomized controlled trials (RCTs), 37 unpublished (FDA data) placebo-controlled RCTs; 86 NSAID versus NSAID RCTs (sample size >or=50); and 103 observational studies. RESULTS: The majority of clinical trials were of good quality. Meta-regression identified an increased risk of dyspepsia for users of specific NSAIDs (adjusted odds ratio [OR] of indomethacin, meclofenamate, piroxicam = 2.8), and for high dosages of other NSAIDs (OR = 3.1), but not for other NSAIDs regardless of dosage (OR = 1.1). Dyspepsia was not reported as an outcome in the case control or cohort studies. CONCLUSIONS: Clinical trial data indicate that high dosages of any NSAID along with any dosage of indomethacin, meclofenamate, or piroxicam increase the risk of dyspepsia by about 3-fold. Other NSAIDs at lower dosages were not associated with an increased risk of dyspepsia. PMID- 12910558 TI - Predicting the onset of forearm pain: a prospective study across 12 occupational groups. AB - OBJECTIVE: To determine, among workers free of forearm pain, the role of mechanical and psychosocial factors in predicting future onset. METHODS: A prospective cohort study was conducted among 782 newly employed workers from 12 occupational groups. At baseline, a cohort of 782 workers free of forearm pain was identified and measurement was made about physical and psychosocial aspects of their job and working environment. Subjects were recontacted after 1 year to determine new onsets of forearm pain. A sample of those reporting new onset forearm pain underwent a structured examination of the upper limb. RESULTS: One year after baseline, 666 (85%) subjects were followed up. The overall prevalence of new onset forearm pain was 8.3% (n = 55). The strongest mechanical risk factor was frequent repetitive movements of the arm or wrist (odds ratio [OR] 2.9, 95% confidence interval [95% CI] 1.6-5.2). The strongest psychosocial risk factors were work considered monotonous at least half of the time (OR 3.0, 95% CI 1.6 5.7) or work with little autonomy (OR 2.6, 95% CI 1.1-6.1). Three specific independent risk factors (monotonous work, repetitive wrist movement, working with hands above shoulder level) could distinguish groups of subjects at substantially different risks of onset. CONCLUSIONS: Along with repetitive movements of the arms and wrists, mechanical postural factors and psychosocial factors also are important risk factors for onset of forearm pain. Our study emphasizes the multifactorial nature of risks for onset of forearm pain, and provides leads as to possible mechanisms for prevention. PMID- 12910559 TI - Socioeconomic impact of osteoarthritis in Hong Kong: utilization of health and social services, and direct and indirect costs. AB - OBJECTIVE: To determine the direct and indirect cost of osteoarthritis (OA) according to disease severity, and to estimate the total cost of the disease in Hong Kong. METHODS: This study is a retrospective, cross-sectional, nonrandom, cohort design, with subjects stratified according to disease severity based on functional limitation and the presence or absence of joint prosthesis. Subjects were recruited from primary care, geriatric medicine, rheumatology, and orthopedic clinics. There were 219 patients in the mild disease category, 290 patients in the severe category, and 65 patients with joint replacement. A questionnaire gathered information on demographic and socioeconomic characteristics, function limitation, use of health and social services, and effect on occupation and living arrangements over the previous 12 months. Costs were calculated as direct and indirect. RESULTS: Low education and socioeconomic class were associated with more severe disease. OA affected family or close relationships in 44%. The average cost incurred as a result of side effects of medication is similar to the average cost of medication itself. Excluding joint replacement, the direct costs ranged from Hong Kong (HK) dollar $11,690 to $40,180 per person per year and indirect costs, HK $3,300-$6,640. The direct costs are comparable to those reported in Western countries; however, the ratio of direct to indirect costs is much higher than 1, in contrast to the greater indirect versus direct costs reported in whites. The total cost expressed as a percentage of gross national product is also much lower in Hong Kong. CONCLUSIONS: The socioeconomic impact of OA in the Hong Kong population is comparable to Western countries, but the economic burden is largely placed on the government, with patients having relatively low out-of-pocket expenditures. PMID- 12910560 TI - The relationship between self-report and performance-related measures: questioning the content validity of timed tests. AB - OBJECTIVE: To examine the determinants of the modest correlation between self report and performance-related measures in patients with osteoarthritis of the hip or knee. METHODS: Measures included the Lower Extremity Functional Scale (LEFS), the self paced walk, timed up-and-go, and stair test. Each performance measure consisted of 3 domains: time, pain (visual analog scale), and exertion (Borg scale). Activity specificity was assessed by examining correlations between the LEFS with single activity and multiple activity time scores. Domain specificity was examined by comparing correlations between the LEFS and single and multiple domain scores. The impact of measurement error was considered. RESULTS: Increasing the number of activity time scores had no effect. Forming a composite performance score based on time, pain, and exertion substantially increased the correlation from 0.44 (composite timed score) to 0.59 (pooled domain and activity score) (P = 0.009). CONCLUSION: Performance scores based on time alone appear to inadequately represent the breadth of health concepts associated with functional status. PMID- 12910561 TI - Efficacy of infliximab in resistant psoriatic arthritis. AB - OBJECTIVE: To evaluate the efficacy and safety of the anti-tumor necrosis factor alpha monoclonal antibody infliximab in the treatment of active psoriatic arthritis (PsA) resistant to previous symptom modifying antirheumatic drugs. METHODS: Sixteen patients with peripheral active PsA with at least 6 months of methotrexate (MTX) therapy at a stable dosage were treated with infliximab administered at a dosage of 3 mg/kg at 0, 2, 6, 14, 22, and 30 weeks while continuing to receive MTX. Intake of nonsteroidal antiinflammatory drugs and corticosteroids was stable during the study period. Standard clinical assessments, erythrocyte sedimentation rate (ESR), and C reactive protein (CRP) were determined at baseline and at weeks 2, 6, 14, 22, and 30. RESULTS: By week 2, significant improvements were registered in the number of swollen and tender joints, visual analog scale for pain, patient and doctor global disease assessment scores, Health Assessment Questionnaire, Dougados functional index, ESR, and CRP. At week 30, the percentages of patients satisfying American College of Rheumatology (ACR) 20%, ACR 50%, and ACR 70% response rates were 64%, 57%, and 57%, respectively. In the 3 patients with active axial disease, spinal stiffness and pain resolved almost completely at week 2 and the improvement did not diminish over time. Psoriasis Area Severity Index improvement was 37% at week 2 and 86% at week 30. No patients dropped out for treatment failure. Side effects were observed in 4 of 16 patients, 2 of whom suspended the therapy due to a severe allergic reaction. CONCLUSION: In patients with resistant PsA, infliximab is an effective therapy without major side effects. PMID- 12910562 TI - sE-selectin for stratifying outcome in rheumatoid arthritis. AB - OBJECTIVES: To determine the usefulness of sE-selectin as a marker for early diagnosis and stratification of rheumatoid arthritis. METHODS: We investigated several markers of disease activity, including circulating adhesion molecules and other standard laboratory tests, in a 2-3 year followup analysis of patients with rheumatoid arthritis. RESULTS: The mean +/- SD levels of sE-selectin (91.68 +/- 31.8 ng/ml versus 49.83 +/- 14.76 ng/ml) and rheumatoid factor (375.7 +/- 394.4 U versus 44.66 +/- 37.63 U) were strongly elevated in severe (n = 15) versus mild (n = 7) courses of disease. Statistical calculation of mean and standard deviation revealed that sE-selectin represents a highly significant marker for the presence of persistent and aggressive disease over time, regardless of therapeutic intervention and observation time points (P = 0.0004). Notably, regression analysis identified constant values for all parameters analyzed and, therefore, a stable course of the disease could be predicted from the beginning. CONCLUSION: sE-selectin appears to be a powerful marker to predict the severity of rheumatoid arthritis. PMID- 12910563 TI - Assessment of depression in rheumatoid arthritis: a modified version of the center for epidemiologic studies depression scale. AB - OBJECTIVE: The Center for Epidemiologic Studies Depression Scale (CES-D) is an instrument commonly used to assess depressive symptoms. Although the psychometric properties of the instrument are well established, the instrument's ability to identify confirmed cases of major depression has been unclear. The purpose of this study was to evaluate the ability of cutoff scores from both a full scale and a modified CES-D to detect major depression in people with rheumatoid arthritis (RA). METHOD: Data were analyzed from 457 persons with RA, including 91 who met criteria for major depression. RESULTS: Results indicated that, in general, a full scale cutoff score of 19 was the most efficient in identifying cases of major depression; the cutoff score of 19 outperformed a variety of other cutoff scores from the modified scale. Even the most efficient cutoff scores, however, demonstrated problems in accurately identifying people with depression. CONCLUSION: The CES-D, while potentially useful as a screening tool, should not be used to identify cases of major depression. PMID- 12910564 TI - Group education for patients with rheumatoid arthritis and their partners. AB - OBJECTIVE: To determine the effects of group education followed by booster sessions for people with rheumatoid arthritis (RA), and to determine whether participation of a significant other influenced the effects. METHODS: A total of 218 RA patients, each of them with a partner, took part in the study. Two-thirds of the patients received a 5-week group self-management education program, with booster sessions after 3, 6, and 9 months; half of them received the intervention with a partner, and half without. One-third of the patients received the same educational materials without group sessions. Data were collected 1 week before the group sessions began and 2, 6, and 12 months later. The assessments included health behavior, arthritis self-efficacy, health status, and social interactions. RESULTS: After 12 months, self-efficacy scores for coping with other symptoms were significantly higher for patients participating in the group education without a partner and significantly lower for patients participating in the group education with a partner. Fatigue increased in patients participating in the group education with a significant other and decreased in patients participating in the group education without a significant other. No other effects were found on health status, health behavior, or social interactions. CONCLUSION: Our findings suggest that participation of a significant other in psychoeducational programs does not have only positive effects. Instead of stimulating patients to adopt beneficial health behaviors and increase their self-efficacy expectations, participation of a significant other led in our program to decreases in self efficacy and increased fatigue, whereas patients participating in group education without partners showed increases in self-efficacy and decreased fatigue. Booster sessions did not seem to influence results. PMID- 12910565 TI - Impact of socioeconomic status on the course of rheumatoid arthritis and on related use of health care services. AB - OBJECTIVE: To quantify the impact of socioeconomic status (SES) among patients with rheumatoid arthritis on 1) health outcomes and related health care utilization in relation to disease duration and 2) changes in health outcomes and related health care utilization over a 2-year period. METHODS: A questionnaire survey was conducted among 878 patients with rheumatoid arthritis (RA), varying in disease duration from 0 to more than 15 years. To determine the impact of SES on the health outcomes and health care use, patients were compared within and between 3 disease duration groups. Additionally, longitudinal changes in health outcomes and health care use were assessed with a followup questionnaire sent out 2 years later. RESULTS: Patients with low SES have worse disease activity, physical health, mental health, and quality of life than patients with high SES. These differences, however, decreased over time. Regarding health care use, we found that patients with low SES made considerably less use of allied health care than patients with high SES. CONCLUSION: Efforts should be undertaken in health care to alleviate the health disadvantages of RA patients in lower socioeconomic groups. In particular, the access to allied health care could be improved. PMID- 12910566 TI - The use of rheumatoid arthritis health-related quality of life patient questionnaires in clinical practice: lessons learned. AB - OBJECTIVE: The utilization of health-related quality of life (HRQOL) patient questionnaires by clinical rheumatologists is limited. Yet, considerable literature exists defining the value of such data. In an effort to understand this apparent paradox, we performed a literature review and conducted a survey to describe what has been learned over the past 2 decades concerning the use of these measures in clinical care and explore the reasons for their underutilization. METHODS: A panel of rheumatologists with extensive clinical experience was convened to review the relevant literature pertaining to the use of HRQOL patient instruments in clinical practice. Additionally, a survey of all American College of Rheumatology practicing clinicians was conducted to assess the use of and beliefs about these measures. RESULTS: The literature provided evidence to support the use of HRQOL patient measures in clinical practice. Forty seven percent of the responding rheumatologists stated that none of their patients complete HRQOL patient questionnaires. The majority of respondents (63%) reported that such information is "somewhat valuable." The most frequently reported reason for the underutilization was that such instruments "require too much staff time." CONCLUSIONS: The literature supports the potential value of HRQOL patient questionnaires in clinical practice. Few rheumatologists routinely gather such information as part of patient care. Reasons for this discrepancy between utility and use are given along with recommendations intended to help increase their utilization in clinical care. PMID- 12910567 TI - Treatment of primary Sjogren's syndrome with low-dose human interferon alfa administered by the oromucosal route: combined phase III results. AB - OBJECTIVE: This study tested the safety and efficacy of human interferon (IFN) alfa for treatment of salivary hypofunction and dry mouth symptoms in primary Sjogren's syndrome patients. METHODS: Combined results are reported from 2 phase III clinical trials in which a total of 497 subjects with primary Sjogren's syndrome received 150 international units of human IFN alfa or matching placebo 3 times per day for 24 weeks by the oromucosal route. RESULTS: Subjects given IFN alfa had a significantly (P = 0.01) greater mean increase in unstimulated whole saliva (UWS) flow, compared with subjects given placebo. In IFN alfa patients, increases in UWS correlated positively and significantly with improvements noted in 7 of 8 symptoms associated with oral and ocular dryness. The coprimary endpoints of stimulated whole saliva flow and oral dryness were not significantly improved in the IFN alfa group relative to placebo. No significant differences were found between the groups with respect to overall adverse event incidence or severity. CONCLUSION: IFN alfa given at low dosage by the oromucosal route can significantly increase UWS flow in patients with primary Sjogren's syndrome, without causing significant adverse events. PMID- 12910568 TI - Clinical and epidemiologic analysis of giant cell (temporal) arteritis from a nationwide survey in 1998 in Japan: the first government-supported nationwide survey. AB - OBJECTIVE: To elucidate epidemiologic and clinical manifestations of Japanese patients with giant cell arteritis (GCA), the first nationwide survey for GCA was conducted in 1998 in Japan. METHODS: The first questionnaire on GCA for patients treated in 1997 was sent to 10,717 medical departments in Japan. A total of 177 patients were reported. Among the 177 patients, 66 GCA patients with detailed clinical and epidemiologic features on second survey were analyzed. RESULTS: Prevalence in patients 50 years of age and older in 1997 was 1.47 per 100,000 population in Japan. The average age at onset was 71.5 years old. The male:female ratio was 1:1.7. The association with permanent and complete visual loss (6.5%), jaw claudication (15.2%), and polymyalgia rheumatica (PMR) (30.3%) were low in frequency compared with those reported from other countries. All patients were treated with corticosteroids. Only 3 (4.5%) patients were reported as deceased due to other causes. CONCLUSION: The prevalence of GCA in Japan was revealed to be extremely low compared with other countries. Clinical findings of permanent and complete visual loss, jaw claudication, and PMR were infrequent among Japanese patients with GCA. PMID- 12910569 TI - Treatment with infliximab for a child with Behcet's disease. PMID- 12910570 TI - Current status of glucosamine therapy in osteoarthritis. PMID- 12910572 TI - Colchicine myopathy in a patient with familial Mediterranean fever and normal renal function. PMID- 12910574 TI - Comment on methotrexate pneumonitis after initiation of infliximab therapy for rheumatoid arthritis. PMID- 12910575 TI - More comments on the "fibromyalgia" label. PMID- 12910577 TI - Basic science research vs. clinical research in cystic fibrosis: Has the pendulum swung too far? PMID- 12910578 TI - Pulmonary mucus: Pediatric perspective. AB - Airway mucus hypersecretion is a clinical feature of a number of childhood diseases, including asthma and bronchitis-associated conditions. However, compared with adults, there is relatively scarce information concerning mucus pathophysiology in respiratory diseases in children. The available evidence indicates many similarities between adult and childhood respiratory hypersecretory conditions, including goblet-cell hyperplasia and submucosal gland hypertrophy, and airway mucus plugging in asthma. Consequently, it is likely that treatments that are effective in adults would be effective in children. Numerous therapeutic targets are linked to the pathophysiology of airway mucus hypersecretion in experimental models and adults with respiratory disease. Whether or not these same targets are relevant in children is for the most part unclear. These targets include the inflammatory cells mediating the inflammatory response that generates the hypersecretory phenotype, and highly specific cellular elements such as epidermal growth factor receptor tyrosine kinase and calcium-activated chloride (CACL) channels. Identification of these factors is linked with the development of different classes of pharmacotherapeutic molecules directed at these targets. Compounds with a broader spectrum of anti-inflammatory activity are likely to be more effective than compounds with restricted activity. However, certain highly specific targets, such as human CACL1 channels, appear to be strongly associated with the development of an airway hypersecretory phenotype. Data from current clinical trials in adults with blockers of these specific targets are awaited with great interest. The hope is that, if effective, pediatric trials with these compounds could be initiated with a view to alleviation of the clinical impact of airway mucus hypersecretion in children. A significant challenge to the therapeutic progression of these new compounds is effective delivery to the airways in children, with the research effort into development of new compounds matched by advances in inhaler design. PMID- 12910579 TI - Methicillin-resistant Staphylococcus aureus in children with cystic fibrosis: An eradication protocol. AB - A retrospective 12-year study (May 1988-July 2000) was undertaken in children with cystic fibrosis (CF) to evaluate 1) the magnitude of methicillin-resistant Staphylococcus aureus (MRSA) in these children; 2) the clinical impact of MRSA on CF; and 3) the efficacy of an MRSA protocol aimed at the eradication of the carrier state. The study maneuver comprised of 1) surveillance cultures of throat/rectum to detect the MRSA carrier state; 2) life-long cephradine rather than flucloxacillin to lift selection pressure; 3) topical application of oral and nebulized vancomycin for 5 days to clear the carriage of MRSA; and 4) a strict antistaphylococcal hygiene program, including handwashing and device policy. Fifteen children with CF (11 boys, with median age 117 months) positive for MRSA were enrolled. The current prevalence of MRSA among children with CF in our hospital is 6.5%. Of 15 children identified, only 12 (18 episodes of MRSA colonization) were treated according to protocol. Median age of MRSA acquisition was 73 months (interquartile range, 43-134 months). In 7 patients (55%), MRSA was eradicated. Of a total of 18 MRSA episodes, the protocol was successful in 10 episodes. The mean period of MRSA-free status was 12 months (range, 6-36 months). Pulmonary function (measured by FEV(1)) was not affected (68% of predicted before treatment, and 68% of predicted after treatment). All children were oropharyngeal carriers of both MRSA and ceftazidime-resistant P. aeruginosa. We believe that an effort has to be made to limit MRSA in CF clinics for the following reasons: 1) MRSA carriage in any individual is an abnormal condition; 2) limitation of systemic vancomycin use is desirable; 3) MRSA carriage may be a contraindication for lung transplantation; and 4) epidemiologically, a CF unit with a substantial MRSA problem functions as a source of dissemination for other patients. PMID- 12910580 TI - Assessing bronchodilator responsiveness in infants using partial expiratory flow volume curves. AB - Our objective was to compare the effectiveness of maximum forced expiratory flow measured at functional residual capacity (V'maxFRC) and the ratio of flow at 75% of the forced expiratory volume to peak forced expiratory flow (FEF(75)/FEF(peak)) for detecting bronchodilator-related changes in wheezy infants. In 55 infants (mean age, 7.8 +/- 3.1 months) with a history of recurrent wheezing, V'maxFRC and FEF(75)/FEF(peak) were measured at baseline and 15 min following nebulized albuterol. Mean results from 4 baseline and 4 postalbuterol partial expiratory flow-volume curves were compared at baseline and following bronchodilator challenge. The strength (relative effect size) of each measure for assessing change was quantified by dividing the mean of the pre- to postdifferences by the standard deviation of the differences. Mean percent predicted V'maxFRC was 41.3 +/- 34.3% at baseline and 44.4 +/- 34.0% following albuterol. Mean FEF(75)/FEF(peak) was 26.7 +/- 13.4% at baseline and 35.8 +/- 14.3% following albuterol. The mean percent change from baseline [(post-pre)/pre] in percent predicted V'maxFRC was 18.3 +/- 39.3, and for FEF(75)/FEF(peak), it was 44.1 +/- 36.8. The change in FEF(75)/FEF(peak) following albuterol was significantly greater than the change in V'maxFRC (P < 0.0001). The relative effect size for mean percent change from baseline in V'maxFRC was 0.47, and for FEF(75)/FEF(peak), 1.20. Changes in FEF(75)/FEF(peak) appear to differentiate changes in airway function following administration of a bronchodilator better than do changes in V'maxFRC. PMID- 12910581 TI - Correlation between standing height, sitting height, and arm span as an index of pulmonary function in 6-10-year-old children. AB - The present study evaluates the relationships of pulmonary volumes and flows and the anthropometric variables of standing height, sitting height, and arm span in 100 6-10-year-old children. To be included in the study, all children were required to be healthy and above the 3rd percentile of the curve of Tanner et al. (Arch Dis Child 1966;41:454-471; Arch Dis Child 1966;41:613-635) for height and weight. Standing height, sitting height, and arm span were measured according to standard protocol. The pulmonary function measurements were: tidal volume, minute respiratory volume, respiratory rate, expiratory reserve volume, inspiratory capacity, vital capacity, residual volume (RV), functional residual capacity, total lung capacity (TLC), forced vital capacity (FVC), forced expiratory volume in 1 sec of FVC (FEV(1)), forced expiratory flow measured between 25-75% of FVC, and FEV(1)/FVC and RV/TLC ratios. Simple linear regression best expressed the correlation between pulmonary function and the anthropometric variables. Significant correlations between anthropometric and pulmonary function measurements were observed, with FVC and FEV(1) showing the highest and RV the lowest r(2) values. There was a significant positive correlation between standing height and arm span according to linear regression and Bland-Altman comparison (Bland and Altman, Lancet 1968;8:307-310). Our results suggest that when an accurate determination of standing height cannot be obtained, arm span could be used interchangeably, using the same regression equation. However, for patients with limb deformities, regression equations of pulmonary function measurements in relation to sitting height may be a better choice for estimating pulmonary function. PMID- 12910582 TI - Migration to a western country increases asthma symptoms but not eosinophilic airway inflammation. AB - The prevalence of asthma symptoms varies markedly throughout the world. However, the asthma mechanisms involved are not defined. Studying the effects of migration can help identify the reasons for this geographic variation. The aims of this study were to examine the prevalence of asthma symptoms, airway hyperresponsiveness (AHR), and induced sputum eosinophils in adolescents who migrate to Australia. The study was conducted in Sydney, Australia, where adolescent students completed a video symptom questionnaire, hypertonic saline challenge, sputum induction, and allergy skin testing. The 211 students had widely different cultural backgrounds, including Asian, South Pacific, Middle Eastern, European, and African countries. Among adolescents who were migrants to Australia, the prevalence of asthma symptoms was higher than that reported using a similar methodology in their country of origin. Asthma symptom prevalence was related to residence time in Australia. The prevalence of wheeze was 17.2% in recent arrivals, 20.5% in adolescents living in Australia for >2 years, and 36.3% in those living all their lifetime in Australia (P = 0.013). For every year of residence in Australia, there was an 11% increase in prevalence of current wheeze (odds ratio, 1.11; P = 0.02). This effect was not related to atopy, AHR, or eosinophilic airway inflammation. Sputum neutrophils were elevated in recent arrivals. In conclusion, adolescents who migrate to Australia report increased asthma symptoms, compared to their country of origin, and asthma symptoms are further increased for every additional year of residence in Australia. The development of wheeze after migration to Australia was independent of eosinophilic inflammation and consistent with noneosinophilic asthma mechanisms. PMID- 12910583 TI - Validity of esophageal pressure measurements with positive end-expiratory pressure in preterm infants. AB - Previous research suggested that esophageal pressure changes (DeltaP(es)) may not reflect pleural pressure changes (DeltaP(pl)) in the presence of positive end expiratory pressure (PEEP), making assessments of dynamic lung mechanics invalid in these circumstances. To test this hypothesis, we measured DeltaP(es) using a water-filled catheter in 18 preterm infants with lung disease (9 intubated), and adjusted the catheter position to achieve a valid occlusion test. End-expiratory occlusions were then carried out at PEEP (cm H(2)O) of 0, 4, and 8, and plots of DeltaP(es) against DeltaP(ao) during airway occlusion were examined to derive the ratio DeltaP(es)/DeltaP(ao) and the r value (as a measure of linearity). There was no significant change in DeltaP(es)/DeltaP(ao), which remained close to 1.0 as PEEP was increased from 0 to 8 cm H(2)O, and r also remained close to unity, indicating no appreciable hysteresis or alinearity of the plots. Our results show that DeltaP(es), when measured with an appropriately placed water-filled catheter, continues to reflect DeltaP(pl) accurately when lung volume is raised by applying PEEP up to 8 cm H(2)O. PMID- 12910584 TI - Anaerobic exercise in pediatric cystic fibrosis. AB - Anaerobic fitness is important for daily functioning of children with cystic fibrosis (CF). The aim of this study was to assess the determinants of anaerobic performance in CF. Anaerobic performance was measured in 39 children with CF (mean age, 13.2 +/- 1.8 (SD) years, forced expired volume in 1 sec (FEV(1)) 81.6 +/- 22.1% predicted), using a Wingate anaerobic test. Significant associations were found for peak power (PP) and mean power (MP) with fat-free mass (FFM) body weight, body mass index, maximal isometric muscle force, and aerobic capacity. Pulmonary function was correlated with anaerobic indices when controlled for FFM. Multiple regression analysis indicated that FFM and FEV(1) accounted for 82% and 86% of the variability in PP and MP, respectively. Patients with moderate CF (FEV(1) < 80%), as compared to mild CF (FEV(1) >/= 80%), had higher PP (difference = 85 W, 95% CI = 27-144 W) and MP (difference = 53 W, 95% CI = 42-63 W) at equivalent FFM. Our results indicate that FFM and pulmonary function are important determinants of anaerobic exercise performance in children with CF. With progression of pulmonary disease, anaerobic performance may be enhanced. PMID- 12910586 TI - Inhaled corticosteroids in childhood asthma: Income differences in use. AB - Asthma hospitalization rates in children increase with decreasing level of household income. This research was undertaken to determine whether use of inhaled corticosteroid drugs, which can prevent asthma hospitalizations, followed a similar socioeconomic gradient in children with asthma. We performed a cross sectional study of association, using population-based prescription and healthcare data sources. Our subjects were 16,862 Manitoba children, aged 5-15 years, with prescriptions for asthma drugs during January 1995-March 1996. Our measures were adjusted for asthma severity, physician specialty, and proportion of children with an inhaled corticosteroid prescription by neighborhood income. Forty-five percent of children treated for asthma had at least one inhaled corticosteroid prescription during January 1995-March 1996. The proportion of children with inhaled corticosteroid prescriptions decreased with successive decreases in neighborhood income. The socioeconomic gradient in the likelihood of an inhaled corticosteroid prescription was most evident among children with mild moderate asthma who were not in the care of an asthma specialist. In conclusion, a socioeconomic gradient in the use of inhaled corticosteroids prescriptions can be found among children with universal access to healthcare and drug insurance. PMID- 12910585 TI - Longitudinal evaluation of bronchopulmonary disease in children with cystic fibrosis. AB - Children with cystic fibrosis (CF) develop bronchopulmonary disease at variable ages. Determining the epidemiology of chronic lung disease and quantifying its severity, however, have been difficult in infants and young children. As part of the Wisconsin CF Neonatal Screening Project, we were presented with an ideal opportunity to assess longitudinally the evolution of symptoms, signs, and quantitative measures of CF respiratory disease. After newborn screening test results led to early recognition, 64 patients diagnosed at a median age of 6.71 weeks were enrolled and studied systematically at a median age of 11.3 years to obtain clinical information, chest radiographs, and pulmonary function tests. Our observations revealed that a frequent cough by history is evident by 10.5 months of age in half the patients. Quantitative chest radiology (CXR scoring) demonstrated that potentially irreversible abnormalities are present in half the children by 2 years. The severity of Wisconsin and Brasfield CXR scores increased in association with respiratory infections. Longitudinal progression of Wisconsin CXR scores was related to age (P < 0.001), pancreatic insufficiency (P = 0.005), and respiratory secretion cultures positive for Staphylococus aureas (P = 0.039). In contrast, serial spirometry showed limited sensitivity, as did lung volume determinations; neither was satisfactory as repeated measures with acceptable quality control until after 7 years of age. Time to event analyses revealed that half the patients had % predicted FEF(25-75) and FEV(1)/FVC values greater than 80% until 10.7 and 9.9 years, respectively. We conclude that of the methods evaluated, quantitative chest radiology is currently the best procedure for frequent assessment of bronchopulmonary disease in CF, and that radiographic progression is evident in approximately 85% of patients by 5 years of age. Our results also suggest that bronchiectasis and other radiographic evidence of chronic infection are apparent prior to airways obstruction in young CF patients. PMID- 12910587 TI - Changes in lung volume and work of breathing: A comparison of two variable-flow nasal continuous positive airway pressure devices in very low birth weight infants. AB - Variable flow nasal continuous positive airway pressure (VF-NCPAP) recruits lung volume more effectively and reduces work of breathing (WOB) compared to constant flow NCPAP (CF-NCPAP) in very low birth weight (VLBW) infants. Because different VF-NCPAP devices have somewhat different flow patterns, whether different VF NCPAP devices function similarly is unknown. We compared two VF-NCPAP devices: the Infant Flow trade mark (EME, Ltd.) and the Arabella(R) (Hamilton Medical) to assess whether lung volume recruitment and WOB were similar in VLBW infants requiring NCPAP. Eighteen infants <1,500 g were studied on both NCPAP devices applied in random order. All infants required NCPAP for mild respiratory distress. Calibrated DC-coupled respiratory inductance plethysmography (RIP) was used to assess lung volume changes. NCPAP was first increased to 8 cmH(2)O to allow comparable recruitment in all infants, and then was slowly decreased to 6, 4, and 0 cmH(2)O, with data collection at each level. Mean birth weight (+/-SD) was 1,107 +/- 218 g, gestational age was 27.9 +/- 2.0 weeks, weight at study was 1,092 +/- 222 g, and age at study was 4.6 +/- 4.3 days. There were no differences in lung volume recruitment overall or at any NCPAP level (P = 0.943). No differences were found in either inspiratory WOB (P = 0.468) or in resistive WOB (P = 0.610) between devices. Compliance, tidal volume, respiratory rate, and minute ventilation were also similar. Despite differences in flow characteristics between the two VF-NCPAP devices we studied, lung volume recruitment and WOB were similar. PMID- 12910588 TI - Congenital pulmonary capillary hemangiomatosis: Report of two cases and review of the literature. AB - Pulmonary capillary hemangiomatosis (PCH) is a rare disease characterized by pulmonary hypertension and excessive neovascularization within the pulmonary interstitium, vasculature, and airways. We describe two unusual cases of congenital PCH. Both cases had concurrent anomalies, including renal and urinary bladder agenesis and hypertropic cardiomyopathy. In one case, capillary proliferation caused significant impingement of the proximal bronchial airways. A review of the current literature is described. PMID- 12910589 TI - Retropharyngeal and mediastinal abscess following adenoidectomy. AB - Adenoidectomy is one of the most common surgical procedures in children. On rare occasions it can be complicated by a retropharyngeal abscess (RA). We report the case of a 9-year-old girl with an RA extending to the mediastinum following adenoidectomy. The mediastinal collection was successfully treated with computed tomography-guided percutaneous drainage to avoid extensive surgical procedure. Surgeons should be aware of these dramatic complications and should avoid overzealous surgery in the removal of the adenoid tissue. PMID- 12910590 TI - [Infections: an inseparable part of medicine]. AB - In review-article personal view of the author on main issues of infectious diseases is presented. Infectious diseases are still very important problem with features of difficult to overcome crisis. It is a result of many biological, social, and political factors. They are an inseparable part of medicine developing together with progress of sciences. PMID- 12910591 TI - [Characteristics of various types of Creutzfeldt-Jakob disease]. AB - The author presents main types of Creutzfeldt-Jakob disease with short description of their ethiopathogenesis, clinical symptomatology, epidemiology and diagnostic criteria. Relevancy of currently applied laboratorial methods in diagnosis of CJD is discussed. PMID- 12910592 TI - [Chlamydia pneumoniae as a biological factor contributing to pathological changes of arterial walls in patients with atherosclerosis]. AB - In this study, using scanning and transmission electron microscope, we attempt to evaluate ultrastructural alterations of endothelial cells, macrophages and smooth muscles cells. The inflammatory process has an essential impact on the development of Chlamydia infection. Specimens from human carotid were obtained from patients who underwent endarterectomy. For examination under scanning and transmission electron microscope vessel sections were fixed in paraformaldehyde and glutaraldehyde. We analysed alterations of endothelial cells covering advanced atherosclerotic plaque in carotid using scanning electron microscope. Smooth muscle cells had undergone the heaviest proliferation among the cells on artery wall. In the tested material we detect diversified morphological forms of Chlamydia sp. We found that one of the pathogens that may lead to atherosclerotic lesions is Chlamydia pneumoniae. PMID- 12910593 TI - [Virulence of Yersinia pseudotuberculosis strains isolated from environment as a threat to human epidemic]. AB - The purpose of the work has been to identify Yersinia pseudotuberculosis strains and to demonstrate their potential pathogenicity using PCR reaction. Investigations included 167 samples of the water and 32 samples of the soil from westpomeranian region. Based on the analysis of PCR reactions the research confirmed the presence of DNA Y. pseudotuberculosis strain in 6 of the water samples and in 1 of the soil sample. The strains have been identified by nucleotide sequence of the ypm gene, which is specific only for mentioned species. Genotypic analysis demonstrated also the presence of genes, which confirmed their virulence. The PCR reaction should be used in the microbiological diagnostic of Y. pseudotuberculosis strains, isolated from animals and from environment, because it is very specific, fast and sensitive method. PMID- 12910594 TI - [Sequelae of tick-born encephalitis in cases treated in the infection observatory ward of Regional Hospital in Gizycko]. AB - The paper reveals the description of the sequels of tick-born encephalitis among patients treated in Infectious Observatory Ward of Country Hospital in Gizycko during 2000-2002. PMID- 12910595 TI - [Pertussis: recurrent threat]. AB - Disturbing increase of pertussis diseases in Poland in the last twelve years has been observed also in Bialystok district and in Bialystok City. Incidence rates exceed an average of morbidity in Poland. Most of illnesses are registered at children between 5 to 9 years old, completely immunized against this disease. There were more females than males. The most of patients needed hospitalization. PMID- 12910597 TI - [Clinical picture of Herpesviridae infections among immunocompromised patients: bone marrow and solid organ transplants recipients]. AB - The human herpes virus (HHV) family (herpesviridae) are large DNA viruses containing eight important, ubiquitous human pathogens. This group of viruses encompasses: herpes simplex virus (HSV types 1 and 2), varicella-zoster virus (VZV), Epstein-Barr virus (EBV), cytomegalovirus (CMV), HHV-6, HHV-7 (cause roseola or exanthema subitum in children) and Kaposi sarcoma herpes virus- (KSHV). The outstanding property of herpes viruses is lifelong persistence of infection and potential periodic reactivation, particularly often among immunocompromised patients. Herpesvirus infections are associated with a wide spectrum of diseases ranging from local ulceration to serious systemic illnessess or malignancies. These infections are one of the major cause of morbidity and mortality in the immunocompromised patients. PMID- 12910596 TI - [Prevalence of antibodies and vaccination against measles, mumps and rubella in Polish population]. AB - The data according number of cases and morbidity rates of measles, mumps and rubella as well as antibody prevalence against diseases above mentioned in Polish population were shown. The impact of mass vaccination on decreasing of measles incidence and number of reported rubella congenital syndrome was observed. The possibility of elimination these diseases in the near future was discussed. PMID- 12910598 TI - [Controversy concerning optimal prophylaxis and empirical antifungal therapy in immunocompromised patients]. AB - This article presents actual major problem about a steady increase in frequency of opportunistic invasive fungal infections (IFIs) in immunocompromised patients. However, there still remains much uncertainty regarding the best methods for establishing the diagnosis of most IFIs. An international consensus, that defining opportunistic IFIs proposed three levels of probability: "proven", "probable", and "possible". Practising physicians approach this uncertainty by prophylaxis and antifungal empirical therapy. Unfortunately, up to now we dispose only few antifungals compounds and all have narrow of therapeutic windows. This article reviews the therapeutic options in chemoprevention and antifungal therapy. Fluconazole and itraconazole are the first durable alternatives to polyenes in chemoprophylaxis. However their use remains controversial as debate continues over both their effectiveness and their potential to select out resistant Candida sp. Amphotericin B is the "gold" standard for the treatment both empirical and proven IFIs, but this drug is frequently associated with severe nephrotoxicity. The lipid formulations of amphotericin B enable higher dosages to be administrated with lower incidences of side effects but its effectiveness is not sufficient. It is to be hoped that rationally designed clinical trials with the new compounds, such as for example echinocandins will lead to improved prevention and treatment of IFIs. PMID- 12910599 TI - [Immune mechanisms in HIV infection and their role in antiretroviral therapy]. AB - In this article mechanisms of CD4 cells depletion, apoptosis, necrosis and T-cell activation, cellular immune responses to HIV infection, with emphasis on involvement of CD4 and CD8 HIV-specific cytotoxic T lymphocytes in human immunodeficiency virus infection are being discussed. Author reviews recent reports on immune restoration resulting from antiretroviral therapy, T-cell turnover within CD4 and CD8 subsets, including rapidly (activated effector and memory) and slowly (naive) proliferating T-cells and the influence of IL-7 on regeneration of T lymphocytes. Finally function of dendritic cells in pathomechanisms of HIV disease is summarized. PMID- 12910600 TI - [Extrahepatic replication of the hepatitis C virus (HCV)]. AB - Pathogenetic aspects and experimental methods employed in the determination of extrahepatic replication of HCV are summarized in the paper. Cells, tissues and organs supporting the active viral replication, particularly cells of the immune system, are presented. PMID- 12910601 TI - [Viral nosocomial infections: the problem of contemporary hospital management]. AB - The most frequent viral pathogens which are the cause of nosocomial infections were presented. Influenza and parainfluenza viruses as well as RS virus affect frequently respiratory tract. So called enteric viruses which are rotaviruses, adenoviruses, small round viruses, astroviruses, caliciviruses, corona viruses, Coxackie, ECHO may be the agents of disorders in digestive tract in the form of intoxications. Viruses of viral hepatitis B, C, D and HIV, CMV, EBV may be the source of nosocomial viral infections transmitted by blood (transfusions). PMID- 12910602 TI - [Occurrence of insect pests in hospitals in Poland]. AB - The prevalent synantropic species present in hospitals in Poland was the German cockroach (Blattella germanica L.), found in about 70% hospitals. It was followed by Oriental cockroach (Blatta orientais L.) and Pharaoh's ant (Monomorium pharaonis L.) occurred in 40% and 17% of hospitals respectively. Kitchens, laundries and lavatories were the most often infested places. Preliminary investigation of German cockroaches caught in hospitals in Poland showed on their body surfaces presence of bacteria known as these causing nosocomial infection. Several strains were resistant to antibacterial drugs widely used for treatment and showed insensitivity to chemical disinfectants used for surface treatment. Additional risk elements in Poland could be high resistance levels to many insecticides used for insect control in hospitals. PMID- 12910603 TI - [Travel medicine]. AB - The risk of the health join to travel are briefly presented. The following factors are mentioned: travel by fly, climate, expose for diseases. The most often pathogens are considered: bacteria, viruses, parasites, and some imported diseases and their vectors. The crucial importance of the stress and conditions of travel is underlined. It has been suggested some procedures which can protect before the risk of travel. PMID- 12910604 TI - [Visceral leishmaniasis]. AB - Leishmanioses are widespread in 88 countries of the tropical and subtropical zone, including regions of the Mediterranean Sea basin of Southern Europe. Actually, approximately 350 million of people live in Leishmania endemic areas and about 12 million of individuals are infected. Visceral leishmaniosis (kala azar disease, tropical splenomegaly) is caused by at least 3 species of Leishmania protozoa: L. donovani, L. infantum and L. chagasi. The incidence of the disease is estimated at 500,000 new cases annually. The infection is transmitted by Phlebotomus or Lutzomyia mosquitos bites, in which intestines forms invasive to humans are developed. Leishmania spp. have a predilection to the reticulo-histiocytary system cells, leading to their proliferation and disruption, and after spreading to the circulation they invade spleen, liver and bone marrow. Visceral leishmaniosis should be suspected in travelers returning from tropical and subtropical areas with signs of splenomegaly and twice temperature spikes in a day. We reported a case of the kala-azar disease in the 22 year-old Polish patient seasonally working in Italy. The clinical picture was expressed by two daily pikes of fever proceeded by chills, excessive sweat, hepatosplenomegaly, lymphadenopathy, general weakness, abdominal pain and nausea. The Leishmania infection was complicated by candidiosis. Laboratory tests showed anaemia, thrombocytopenia, leucopenia, hypergammaglobulinaemia and a suppression of immunological cellular response. The diagnosis was confirmed by a presence of amastigota forms in macrophages of the bone marrow aspirate and a detection of specific antibodies to L. infantum by Westernblotting. The patient was successfully treated with Glucantime. PMID- 12910605 TI - [Bioterrorism threat]. AB - In this article contemporary views on terrorism with biological weapons use are presented. The bioterrorism phenomenon is described; some of the attacks known so far are reviewed as well as sources of possible agents' acquisition, terrorists' capabilities and lines of conduct. PMID- 12910606 TI - [Animal pathogens as biological weapons]. AB - In this article, the most important animal pathogens, which, according to CDC classification, can be used as biological agents, are presented. The means of dissemination and the ways of infection's propagation are reviewed. Typical and the most dangerous signs and symptoms are described, as well as the consequences of these infections. PMID- 12910607 TI - [Protection against bioterrorism acts in Poland]. AB - In this article general principles of defense against bioterrorism are presented. Furthermore, the effort, already undertaken for organization of the system enabling effective countermeasures to this threat as well as liquidation of the effects of such an event, is shown. PMID- 12910608 TI - [Methods of biological weapon threats detection]. AB - Detection and identification of biological weapon agents are ones of more important elements of defence against biological weapon and bio-terrorism. So, the attack agent determination creates the chance of necessary prophylactic- medical and liquidation activities undertaking on time. Different systems of biological hazards detection are known. Among others, they allow to detect the biological agents presence in the air, execute the aerosol particles analysis, and are able to generate the alarm signal (LRBSDS, JBSDS, FLAPS). Some systems are able to detect biological agents presence, and moreover, to identify them (JPS, JBAIDS). Luminometric method possesses many advantages, from which the versatility of its usage at different attack scenarios, is getting to stay the most important one. A lot of attention is paid to identification problems, recently. Simple technologies, adjusted to field conditions, which give initial identification result within several minutes (immuno-chromatographic tests), have been developed. These tests, based on colloidal gold may be replaced with UCP particles, what influences positively for identification sensibility and specificity. Systems allowing to execute genetic tests in field conditions (RAPID) and biosensors, thank to which bacteria and their toxins, and viruses can be identified (RAPTOR) have occurred. Huge diagnostic possibilities are created by flow cytometry. Instrumental techniques, among which the mass spectrometry pays attention, are developed. Huge hope is connected to systems working on "chips", allowing, among others, to execute significant amount of analyses simultaneously. Thank to huge expenditures, paid for biological agents identification methods development, part of them, after positive laboratory and field practical tests, shall be introduced into the microbiological practice, staying the valuable contribution of fight against bio-terrorism. PMID- 12910609 TI - [Advances in microbiological diagnosis of infectious diseases]. AB - Development of new methods in microbiological diagnosis of infectious diseases is described. Particularly methods which are based on techniques DNA microarray, protein microarray and nanotechnology. PMID- 12910610 TI - [Molecular methods in the diagnosis of infectious diseases]. AB - In the long term, many of the conventional diagnostic approaches to detection and characterisation of infectious diseases is complemented or even replaced by recognition of DNA/RNA specific sequences. The molecular procedures are based on two distinct types of methods--nucleic acid hybridization techniques with specific molecular probes, and DNA amplification by the polymerase chain reaction PCR. In the first method DNA fragments generated by restriction endonucleases are separated by electrophoresis, transferred to proper membrane and annealed with specific oligonucleotides which are labelled with 32P or with a non-radioactive marker. PCR reaction utilizes a DNA extension enzyme (polymerase) which can add nucleotide bases to primers once a template is provided. Oligonucleotides primers have to recognize target molecule. The single cycle is then repeated and each time DNA segments are doubled. Multiplex polymerase chain reactions have been designed to identify an organism as well as its virulence-specific sequences or antibiotic resistance plasmids simultaneously. The major drawback of PCR procedure which result from its exquisite sensitivity have to be consider. The problem is contamination and many strategies have been developed to avoid it. To these research tools were added probes for in situ hybridization. Advantages provided by this technique include the ability to detect latent (non-replicating) viruses and to localize their genomes to nuclear or cytoplasmic regions within cells. Nucleic acid probes or the hybridization conditions can be manipulated so that a broad spectrum of genotypes could be detectable. This is particularly valuable in those emerging infections where the individual serotypes are unknown. Recombinant DNA approaches have now been described for detection of a wide range of infectious agents. Some remain research activities, others are more appropriate to the routine diagnostic laboratory. An understanding of a pathogen's life cycle and the host's responses to the infectious agent is enhanced by characterisation of the former's genome using molecular technology. The spread of epidemics or hospital-acquired (nonsocomial) infections is followed and characterised with more accuracy by the identification of unique DNA fingerprints for individual pathogens. The prospects for molecular medicine in microbiology are vast and will have profound effects in laboratory and clinical practice. PMID- 12910611 TI - [Immunoglobulins and rules of passive-active prophylaxis]. AB - This review deals with the safety uses and mechanisms of action intravenous immunoglobulin preparations (IVIG) in prophylaxis and treatment of viruses and bacterial infection and also about the role of immunoglobulins in autoimmune disorders. The second part contains suggested intervals between immunoglobulin administration and measles immunization. PMID- 12910612 TI - [Evaluation of selected parameters of blood coagulation and fibrinolysis system in patients undergoing total hip replacement surgery with normovolemic hemodilution procedure and standard enoxaparine prophylaxis]. AB - The aim of the study was to evaluate selected blood coagulation and fibrinolysis parameters in patients undergoing total hip replacement surgery with normovolemic hemodilution and standard enoksaparine profilaxis. The study included 66 patients undergoing hip replacement surgery. The group consisted of 51 women and 15 men, within the age range of 47-78, the mean age was 64. In 32 (subgroup II) patients the surgery was performed with the use of normovolemic hemodilution, in 34 (subgroup I) the hemodilution procedure was not applied. The enoksaparine as prophylaxis started 12 hours prior to surgery and continued during hospitalisation. The examination of the coagulation system was performed: on the day of the operation in the morning, on the day of the operation in the evening and on the first day after operation. We determined the concentrations of TAT and PAP complexes, prothrombin fragments 1 + 2 (F1 + 2) and d-dimers (DD). CONCLUSIONS: 1) during total hip replacement surgery and particularly in the period of the first 12 hours after the procedure marked activation of coagulation and fibrinolysis occurRed; 2) the application of the hemodilution procedure does not influence significantly the degree of coagulation and fibrinolysis disorders in the perioperative period, but could reduced incidence of thromboembolic complications in the postoperative period. PMID- 12910614 TI - [Intra- and postoperative complications in the arthroscopy of the knee joint]. AB - Arthroscopy of the knee joint is one of the invasive surgical procedures that are connected with a relatively low risk of complications. Currently it is considered that in the group of diagnostic arthroscopies and technically simple operative arthroscopies all possible complications (less and more serious) account for 0.8 1.2% and in technically more advanced operations performed by means and under control of arthroscopy--1.5-5%. The purpose of this study was a retrospective analysis of intra- and postoperative complications in 10,770 arthroscopies performed in the years of 1986-2001. In our material a number of 731 (6.98%) various complications was observed. General complications (anaestesiological and cardiovascular) represented 0.15%, intraoperative complications--0.31% and postoperative complications--6.34%. PMID- 12910613 TI - [Graft remodeling following Hertel ACL reconstruction in MR imaging]. AB - MRI and clinical assessment of 21 patients operated on with Hertel method due to knee instability has been presented. Position of the graft in reference to the Blumensaat line has been evaluated in MRI with the use of 3 point scale after mean follow-up of 9-months. Four points scale has been used to grade signal intensity as indicator of graft remodeling. Lysholm scale served for clinical assessment. Mean grade for graft position was 2.3 and for signal intensity 2.7. Better remodeling of the graft correlated positively with its better positioning, but clinical assessment correlated poorly with signal intensity and localization of the graft. Incorrect positioning of the graft hinders its remodeling- ligamentization of the tendon. PMID- 12910615 TI - [Early diagnosis of post-traumatic algodystrophy]. AB - The results of the retrospective analysis of the chronology and sequence of symptoms and signs occurrence in 64 patients who developed algodystrophy as a consequence of trauma/operation are presented. The following features were noted: diffuse pain, swelling, reduction of movement of fingers, temperature and colour changes of the limb. Basing on these data, we determined how early after accident a set of features sufficient to establish the diagnosis could be detected. It was found that diagnosis of algodystrophy could be made at 1 week in 22 patients (34%), at 2 weeks in 37 (58%), at 3 weeks in 46 (72%) and at 4 weeks in 53 patients (83%). Only in 11 patients (17%) algodystrophy appeared later than 1 month after initial trauma. The syndrome has been eventually diagnosed at 1 month only in 20 patients (31%). A traditional approach to the algodystrophy as a chronic pain syndrome caused probably considerable delay in the diagnosis, which occurred in almost 70% of our patients. The results of the study show that algodystrophy can be diagnosed in most cases as early as at 1 month after initial trauma. Early diagnosis prevents of development of florid condition and allow the patients to recover in relatively short time. PMID- 12910617 TI - [Galveston technique in treatment of paralytic spine deformities]. AB - The author's present their own experiences with the Galveston technique in treatment of paralytic spine deformities. Nineteen patients (6 boys and 13 girls) were treated (between 1996 and 2002) at the Swiebodzin Spine Centre and the Special Spine Surgery Unit in Poznan. All patients were unable to sit straight without external support, due to neurologic muscle weakness and pelvic obliquity. Surgical treatment lead to a marked improvement of the spinal deformity and also allowed the patients to sit-straight, without need of any support. PMID- 12910616 TI - [First dorsal metacarpal flap in compound multi tissues thumb reconstruction]. AB - In the years 1985-2000 two females and eleven males aged 3-48 (mean age 27) were treated using neurovascular first dorsal metacarpal flap because of compound thumb multi-tissue defects. In two cases the flap included a fragment of vascularized bone from the index proximal phalanx. Simultaneously a graft of non vascularized bone in three cases, nerve grafts in two cases and a reconstruction of A1 pulley of the FPL tendon in three cases were carried out. Two patients were operated immediately after the injury and the others had delayed reconstructions. In 6 cases the defects resulted from crushing or lacerated wounds of the proximal thumb and in three cases of the distal thumb. Three thumbs were distally amputated. A 3 year-old child had an extensive palmar tissues necrosis due to an electric burn. Pedicled flaps were used in twelve patients and one patient had an island flap. The flaps ranged from 2.24 to 12.0 square centimetres. The donor wounds were directly closed in 6 patients and in 7 patients they were skin grafted. All flaps survived completely resulting in good function. Vascularized bone grafts raised in the flaps healed after 6 and 10 weeks respectively, those non-vascularized from the olecranon healed after 8 weeks and all were further rebuilt giving strong thumb of proper length. 2PD was 8-10 mm. Reconstruction of the sheath elements with flap tissues successfully supported FPL tendon. None of the donor site were observed. PMID- 12910618 TI - [Operative treatment of sacral tumors]. AB - The paper presents posterior access to sacrum according to Gennari used for resection of sacral tumors. Technique was compared with other operative techniques described in literature. The advantage of this method is the low risk of postoperative infection and limited operative technique. PMID- 12910619 TI - [Osteogenesis imperfecta]. AB - The paper describes patients with osteogenesis imperfecta (oi). It gives the causes of oi, shows types according to Sillence. Discuss clinical and radiological appearance of the patients. It provides the latest information about rehabilitation and surgical treatment (multilevel osteotomies and rodding), and supplementation of osteoporosis connecting with oi. PMID- 12910620 TI - [Total elbow arthroplasty]. AB - The authors describe indications and contraindications for total elbow arthroplasty. The brief characteristic of total elbow replacement is done. The authors describe its own case of clinical use of the Coonrad-Morrey implant. PMID- 12910621 TI - [Early results of surgical treatment of dysplastic hip arthrosis with CDH prosthesis]. AB - The paper presents the early results of surgical treatment of osteoarthritis secondary to hip dysplasia with CDH hip prosthesis. Between 2001 and 2002 fourteen cases of hip arthritis secondary to dysplasia were treated with CDH prosthesis. Surgery was performed in women, age ranging from 22 to 52 years (average age: 40 years). In 8 cases a cementless CDH femoral component was used. In all cases cementless Taperloc (Biomet) acetabulum components were used. Weight bearing was allowed 7-14 days post-op. Final patient assessment was done 6-26 months post-op, using the Merle d'Aubigne classification. Patient satisfaction was also taken into account. Functional assessment using this classification yielded 7 points (range: 5-9 points) pre-op, and 11 points post-op (range: 10-12 points). In the post-op assessment, patients reported less pain and an increased range of motion of the hip. No infections were noted. In 1 case deep rein irritation was noted on the second day post-op. In 10 cases there was no limb length discrepancy post-op, while in 4 cases it ranged from 1.5 cm to 3.0 cm. CDH prosthesis allows secure hip replacement procedure, especially in cases with narrow femoral canals. PMID- 12910622 TI - [Relationships between the brain and the immune system]. AB - The concept that the brain can modulate activity the immune system stems from the theory of stress. Recent advances in the study of the inter-relationships between the central nervous system and the immune system have demonstrated a vast network of communication pathways between the two systems. Lymphoid organs are innervated by branches of the autonomic nervous system. Accessory immune cells and lymphocytes have membrane receptors for most neurotransmitters and neuropeptides. These receptors are functional, and their activation leads to changes in immune functions, including cell proliferation, chimiotactism and specific immune responses. Brain lesions and stressors can induce a number of changes in the functioning of the immune system. All these changes are not necessarily mediated by the neuroendocrine system. They can also be dependent on autonomic nerve function. The communication pathways that link the brain to the immune system are normally activated by signals from the immune system, and they serve to regulate immune responses. These signals originate from accessory immune cells such as monocytes and macrophages and they are represented mainly by proinflammatory cytokines. Proinflammatory cytokines produced at the periphery act on the brain via two major pathways: (1) a humoral pathway allowing pathogen specific molecular patterns to act on Toll-like receptors in those brain areas that are devoid of a functional blood-brain barrier, the so-called circumventricular areas; (2) a neural pathway, represented by the afferent nerves that innervate the bodily site of infection and injury. In both cases, peripherally produced cytokines induce the expression of brain cytokines that are produced by resident macrophages and microglial cells. These locally produced cytokines diffuse throughout the brain parenchyma to act on target brain areas so as to organise the central components of the host response to infection (fever, neuroendocrine activation, and sickness behavior). PMID- 12910623 TI - [Interactions between the immune and neuroendocrine systems: clinical implications]. AB - The endocrine and immune systems are interrelated via a bidirectional network in which hormones affect immune function and, in turn, immune responses are reflected in neuroendocrine changes. This bidirectional communication is possible because both systems share a common "chemical language" that results from a sharing of common ligands (hormones and cytokines) and their specific receptors. Cytokines are important partners in this crosstalk. They play a role in modulating the hypothalamo-pituitary-adrenal (HPA) axis responses at all three levels: the hypothalamus, the pituitary gland and the adrenals. Acute effects of cytokines are produced at the central nervous system level, particularly the hypothalamus, whereas pituitary and adrenal actions are slower and are probably involved during prolonged exposure to cytokines such as during chronic inflammation or infection. Several mechanisms have been proposed by which peripheral cytokines may gain access to the brain. They include an active transport through the blood-brain barrier, a passage at the circumventricular organ level, as well as a neuronal pathway through the vagal nerve. The immune neuroendocrine interactions are involved in numerous physiological and pathophysiological conditions and the interactions with the HPA axis may represent a mechanism through which the immune system, by stimulating the production of glucocorticoids, avoids an overshoot of inflammatory response. They may also be involved in the state of hypogonadism, of hypothyroidism and growth inhibition which can occur during inflammatory and infectious diseases. The crosstalk between the immune and endocrine systems is important to homeostasis, since the interactions can produce various appropriate adaptative responses when homeostasis is threatened. PMID- 12910624 TI - [Endocrine markers of cellular immunity: defining the endocrine phenotype]. AB - The term neuroendocrine has been used to define cells that secrete their products in a regulated manner, in response to a specific stimulus. The neuroendocrine system includes neurons and endocrine cells sharing a common phenotypic program characterized by the expression of markers such as neuropeptides, chromogranins, neuropeptide processing enzymes SPC2 and SPC3 (subtilase-like pro-protein convertases) or dense core secretory granules. Various theories such as the APUD (amine precursor uptake decarboxylation) concept, the diffuse neuroendocrine system (DNES) or the paraneuron concept have been put forth to classify neuroendocrine cells as a cohesive group. Neuroendocrine characteristics have been used as evidence of a common embryological origin for normal and neoplastic cells. However, it is now recognized that neuroendocrine characteristics can be observed in various cell types, such as immunocytes, that do not share a common embryological origin with either neurons or endocrine cells. We propose to redefine previous "neuroendocrine" concepts to include the notion that activation of specific genetic switches can lead to the expression of a partial or full neuroendocrine phenotype in a variety of cell types, including immune cells. PMID- 12910626 TI - [The immune status of Schwann cells: what is the role of the P2X7 receptor?]. AB - The peripheral nervous system (PNS) displays structural barriers and a lack of lymphatic drainage which strongly limit the access of molecules and cells from the immune system. In addition, the PNS has the ability to set up some specific mechanisms of immune protection to limit the pathogenicity of inflammation processes following insults by pathogens or inflammatory autoimmune diseases like the Guillain-Barre syndrome. Schwann cells are among the most prominent cells which can display immune capabilities in the PNS. Numerous in vitro studies have shown that Schwann cells were indeed able to display a large repertoire of properties, ranging from the participation to antigen presentation, to secretion of pro- and anti-inflammatory cytokines, chemokines and neurotrophic factors. In vivo studies have confirmed the immune capabilities of Schwann cells. The aim of this review is to present how Schwann cells can participate to the initiation, the regulation and the termination of the immune response in the light of the recent discovery of the Schwann cell expression of purinergic P2X7 receptors. PMID- 12910625 TI - [Astrocytes, cells involved in neuro-immune interactions in the central nervous system]. AB - The astrocyte, the major glial cell in the central nervous system, may influence many aspects of inflammation and immune reactivity within the brain. We have established a model of chronically activated T lymphocytes, interacting with neural cells of diverse origin to study the complex immune regulatory system suspected to lead to neuroinflammatory diseases. We show that human astrocytes became reactive following T cell contact, secreting proinflammatory cytokines, matrix metalloproteinases (MMP) and tissue inhibitors of metalloproteinase (TIMP). The altered MMP/TIMP system was shown to be involved in deleterious effects displayed by activated T cells towards human multipotent neural precursers by controlling their sensitivity to T cell-induced Fas-mediated apoptosis. MMP/TIMP was suspected to stabilize Fas at the cell membrane. In a model of mixed rat glial cells in primary culture (astrocytes, oligodendrocytes), activated T lymphocytes induced the collapse of processes and the death of immature oligodendrocytes. These effects were associated with upregulation of Fas at the cell surface of oligodendrocytes and secretion of MMP and TIMP by astrocytes. By amplifying the expression of inflammatory molecules including the MMP/TIMP system, astrocytes appear to be a crucial relay in the deleterious molecular cascade triggered by activated T lymphocytes. Detection of altered MMP/TIMP in patients suffering from myelopathy associated with retroviral infection (HTLV-1) strongly suggests its involvement in the physiopathological process of the disease. PMID- 12910628 TI - [Matrix metalloproteinases and their inhibitors, modulators of neuro-immune interactions and of pathophysiological processes in the nervous system]. AB - The matrix metalloproteinases (MMPs) belong to a growing family of Zn2+-dependent endopeptidases, secreted or membrane-bound (MT-MMP), that regulate or degrade by proteolytic cleavage protein components of the extracellular matrix, cytokines, chemokines, cell adhesion molecules and a variety of membrane receptors. MMP activity is counterbalanced by their physiological inhibitors, the tissue inhibitors of MMPs (TIMPs), a family of 4 secreted multifunctional proteins that have growth promoting activities. In physiological conditions MMP activity is tightly regulated and altered MMP regulation is associated with pathological processes including inflammation, cell proliferation, cell death and tissue remodeling. The MMP/TIMP system is involved in the development and function of cells of the immune system by promoting their differentiation, activation, migration across basement membranes and tissues. In the last years, data has accumulated indicating that the MMP/TIMP system is expressed in the nervous system where it regulates neuro-immune interactions and plays a major role in pathophysiological processes. In this review, we present recent in vivo and in vitro studies that highlight the contribution of the MMP/TIMP system to various diseases of the nervous system, involving blood brain barrier breakdown, neuroinflammation, glial reactivity, neuronal death, reactive plasticity, and to developmental and physiological processes including cell migration, axonal sprouting and neuronal plasticity. This review also alludes to the beneficial effects of synthetic MMP inhibitors in different animal models of neuropathology. In all, a further understanding of the role of MMPs and TIMPs in the nervous system should contribute to unravel mechanisms of neuronal plasticity and pathology and set the basis of new therapeutic strategies in nervous system disorders based on the development of synthetic MMP inhibitors. PMID- 12910627 TI - [Cytokines and nutritional disorders]. AB - During an infection, a decrease in food intake together with elevated energy expenditure appears. Anorexia is one of the most common signs of illness and is often considered as an undesirable manifestation of sickness. However, compelling data demonstrate that anorexia constitutes an adaptative strategy systematically organised for pathogens elimination. Microbial products stimulate the production by immunocompetent cells of cytokines, which orchestrate the immune response. Since the administration of cytokines reduces food intake, it has been suggested that these agents play a key role in mediating anorexia during infection. This review details the mechanisms of cytokine-induced anorexia, focusing on the role of endogenously produced brain cytokines and more particularly interleukin-1 (IL 1). De novo synthesis of IL-1 occurs in the brain during peripheral infection mimicked by the administration of bacterial endotoxin lipopolysaccharide (LPS). Centrally produced IL-1 acts on its receptors to mediate anorexia as demonstrated by the use of knockout mice and specific IL-1 receptor antagonist. Functional neuroanatomy demonstrates further that LPS or IL-1 specifically activates the hypothalamic neurons that control food intake. Leptin is tightly regulated by IL 1, suggesting the involvement of this hormone in the anorexia of infection. The mechanisms by which hypothalamic arcuate nucleus neuropeptides, which are regulated by IL-1 and leptin, could mediate anorexia during infection are discussed. PMID- 12910629 TI - [Does transforming growth factor-beta (TGF-beta) act as a neuroprotective agent in cerebral ischemia?]. AB - Necrosis and apoptosis are the two fundamental hallmarks of neuronal death in stroke. Nevertheless, thrombolysis, by means of the recombinant serine protease t PA, remains until now the only approved treatment of stroke in man. Over the last years, the cytokine termed Transforming Growth Factor-beta 1 (TGF-beta 1) has been found to be strongly up regulated in the central nervous system following ischemia-induced brain damage. Recent studies have shown a neuroprotective activity of TGF-beta 1 against ischemia-induced neuronal death. In vitro, TGF beta 1 protects neurons against excitotoxicity by inhibiting the t-PA-potentiated NMDA-induced neuronal death through a mechanism involving the up-regulation of the type-1 plasminogen activator inhibitor (PAI-1) in astrocytes. Altogether, these observations suggest that either TGF-beta signaling or TGF-beta 1-modulated genes could be good targets for the development of new therapeutic strategies for stroke in man. PMID- 12910630 TI - [Neuro-immune interactions in psychopathology with the example of interferon alpha-induced depression]. AB - The use of interferon (IFN)-alpha for the treatment of viral diseases or cancers is associated with neuropsychiatric side effects in a large number of patients. The mechanisms by which cytokines induce these symptoms, as well as the vulnerability factors for these effects, have not been yet fully elucidated. Systematic clinical studies, combining biochemical approaches, functional brain imaging and treatment intervention, have been initiated to better understand the phenomenology, pathophysiology, and preventive strategies of the neuropsychiatric effects of IFN-alpha in patients with malignant melanoma or chronic hepatitis C. The findings indicate differential phenomenology and treatment responsiveness of the neurovegetative and mood/cognitive symptoms induced by IFN-alpha, suggesting distinct underlying mechanisms. Impaired neuroendocine function, fronto-striatal dysfunction and decreased monoamine were found to contribute to the pathophysiology of core symptoms of IFN-alpha-induced depression, including symptoms of mood alterations, cognitive dysfunction, anhedonia and psychomotor retardation. In addition, some behavioral and biological markers of the vulnerability for IFN-alpha-induced depression were identified. These findings provide important information concerning the relationship between cytokines and depression. PMID- 12910631 TI - [Genetic control of cardiac tube formation in Drosophila]. AB - In Drosophila, the heart is composed of a simple linear tube constituted of 52 pairs of myoendothelial cells which differentiate during embryogenesis to build up a functional mature organ. The cardiac tube is a contractile organ with autonomous muscular activity which functions as a hemolymph pump in an open circulatory circuit. The cardiac tube is organized in metamers which contain six pairs of cardioblasts per segment. Within each metamer the cardioblasts express a combination of genetic markers underlying their functional diversity. For example, the two most posterior cardiac cells in segments A5 to A7 differentiate into ostiae which allow the inflow of hemolymph in the tube. An additional axial information along the anteroposterior axis orchestrates the subdivision of the cardiac tube into an "aorta" in the anterior region and a "heart" in the posterior region which behave as distinct functional entities. The major pacemaker activity is located in the most caudal part of the heart. This analysis has being made possible by the identification and the utilization of specific morphological and genetic markers and an in vivo observation of cardiac function in the embryo. Functional organogenesis of the cardiac tube is accurately controlled by genetic programs that have been in part identified. Hox genes are responsible for the axial subdivision of the tube into functional modules. They activate, in their specific domains of expression, target genes effectors of the terminal differentiation. On the other hand, part of the information required for segmental information is provided by Hedgehog, a morphogen secreted by dorsal ectoderm, whose activity triggers the ostiae formation in the heart domain. PMID- 12910632 TI - [Implications of environmental cues in cardiomyogenesis]. AB - The staging of murine cardiomyocyte specification and determination was investigated in cultures of tissue explants from pre- and postgastrulation embryos and after transplantation of cardiac or cardiogenic tissues from mouse embryos into chick embryos. The development of cultured and transplanted cells in cardiomyocytes was evaluated by testing the expression of several cardiac transcription factor genes (Nkx 2.5, eHAND, dHAND, GATA 4), alpha cardiac actin, and beta myosin heavy chain protein. In vitro analyses showed that cells with the potential to form cardiac muscle were present prior to gastrulation in 6.5-day postconception (dpc) epiblasts. Although, as shown by in vivo experiments, neurectodermal derived structures did not influence cardiogenesis in epiblast transplants, these transplants did not exhibit full cardiogenic cell differentiation in the chicken environment. In in vitro culture, the neurectoderm also had no effect on murine cardiomyogenesis. In contrast, the presence of endoderm in explants between mid- and late streak stages stimulated emerging mesodermal cells to adopt a myocardial pathway. Mesoderm from late streak explants (7.5 dpc) was capable of differentiating into a cardiac phenotype in the avian heterotopic environment, indicating that the specification of cardiac precursors became irreversible around the late streak stage in mouse embryo. PMID- 12910633 TI - [The mouse as a model of heart morphogenesis in mammals: origin and lineage of myocytes]. AB - In order to follow cardiac precursor cells, we have adopted a retrospective clonal approach, based on the nlaacZ genetic label. Random clones were generated and observed at different developmental stages in murine myocardium. The distribution of these clones in clusters suggest for the first time that cells fated to form myocardium proliferate in two steps. The first growth phase, before E8.5, is dispersive and polarised along the axis of the primitive cardiac tube, contributing to its elongation. The second growth phase is coherent and polarised differentially in different cardiac subregions. Interestingly, this can be correlated with production of geometrical forms (dilatation of a sphere, enlargement of a tube), showing the relation between heart morphogenesis and the controlled proliferation of myocardial cells. The restricted distribution of clones to the right or left ventricule was also investigated with the goal of establishing the time at which cardiac chamber identity emerges. Right and left ventricular lineages appear to segregate early, in agreement with the existence of two populations of cardiac precursors, the so-called primary (or posterior) and secondary (or anterior) heart fields. PMID- 12910634 TI - [The mouse as a model for heart morphogenesis in mammals: the origin of myocytes and studies with cardiac explants]. AB - The heart is one of the first organs to form during embryogenesis since its circulatory function is critical from early stages for embryo survival. In man, morphological events are affected by molecular perturbations, which can lead to a congenital heart defect. It is important therefore to understand not only the molecular signals, but also the morphological events, which govern myocardial cell identity. The study of transgenic mouse lines, Mlc1v-nlacZ-24 and Mlc3f nlacZ-2, has led to the identification of a new precardiac territory, the anterior heart field, which has also been described recently in birds, and which contributes to the myocardium of the arterial pole of the heart. The use of explant cultures also indicates that pharyngeal mesoderm participates in the formation of the outflow tract and right ventricle and shows that the primitive heart tube has a predominantly left ventricular identity. We have also shown that Fgf-10 is expressed in the anterior heart field, where a role for FGF signaling in arterial pole morphogenesis is suggested by inhibitor experiments. Finally explant cultures have been employed to examine the acquisition of left-right atrial identity. The Mlc3f-nlacZ-2 line, which marks the right atrium, allowed us to determine the time window during which left-right signaling confers left-right atrial identity. PMID- 12910635 TI - [Role of molecular biology in understanding the formation of congenital cardiomyopathies in humans]. AB - New insights into the genetics of congenital heart diseases in human beings have been obtained these past years. The identification of genes for heart defects have led to a new clinical approach of these malformations in children and their families. These progresses have been made by means of positional cloning as well as through the analysis of mouse models. These findings uncovered a new complexity in the development of cardiac defects and led to revise the different classifications for congenital heart defects. Pediatric cardiologists have also improved their efficiency in defining cardiac phenotypes in affected individuals and in pedigrees with recurrent malformations. At the present time, clinical consequences of these findings remain sparse, though in particular cases genetic counselling has been dramatically improved. PMID- 12910636 TI - [Pathogenetic mechanisms of the initiation of systemic inflammatory response syndrome (a literature review)]. PMID- 12910637 TI - [Congenital defects of post-heparin lipoprotein lipase. Diagnosis of familial combined lipoproteinemia]. PMID- 12910638 TI - [Comparison of the trypsin inhibitor activity in urine of donors and patients with kidney diseases]. AB - The general and acid-stable activities, i.e. anti-tryptic activity (ATA) and anti chemotryptic activity (ACTA), of a trypsin inhibiter in the urine (TIU) were determined for patients and donors. A lack of reliable differences between the general and acid-stable activities in donors and in patients with nephritis is indicative of that TIU is the main inhibiter of trypsin in human urine. A correlation degree between the general ATA and ACTA (r > 0.7) means that the two domain inhibiter form is excreted into the urine. A comparison between the general and acid-stable ATAs suggests the presence of other trypsin inhibiters in the urine of patients. PMID- 12910639 TI - [The role of peroxidation in pathogenesis of varicose veins]. AB - The role of lipid peroxidation in varicose disease (VD) was studied and clinically substantiated in case of 32 patients with a compensated derangement of the regional phlebohemodynamics. An oxidant-antioxidant misbalance was detected, which displayed itself through a higher level of oxiradicals in the blood plasma as well as through a reduced general antioxidant activity in erythrocytes and a reduced level of enzymes, i.e. superoxide dismutase and catalase, by 40% as compared to the control values in healthy persons. The study results point at an important role of the oxidative stress developing in the capillary angiogenesis and varicose transformation of the surface veins in the legs at a muscle load in the vertical position, if there is constitutive insufficiency of the anti-oxidant protection. PMID- 12910641 TI - [Management principles and experience in the clinical diagnostic laboratory (lecture, part 2)]. PMID- 12910640 TI - [Sickle cell anemia (HBSS) in an adult patient from Tanzania]. AB - There is a description of an observation of patient N., aged 29, (a student of Russian University for Peoples' Friendship and a citizen of Tanzania) suffering from sickle-cell anemia--SCA--(HbSS), homozygous type, which was detected at screening of abnormal Hb by electrophoresis on the cellulose acetate strips. The results of clinical-and-biological examinations of the patients are described. The patient had anemia (Hb 98-83 g/L); thrombotic (pain) crises with dark urination were periodically observed. HbS accounted for the bulwark (98%) of Hb, while small fractions of Hb (HbF and HbA2) were within the normal limits. This was a comparatively mild form of the disease; the patient had minor changes in the heart and lungs with other internal bodies being intact. Patient N. received, from childhood, an appropriate therapy including antibiotics, folic acid, analgesics etc. Obviously, this provides for an explanation to that he lived up to 29 years, withstood the acclimatization in Russia and could get education. A comparison of the clinical-and-laboratory indices of patient N. with the data of patients with SCA of other types (HbS-thalassemia, HbSC and HbSK) is presented. PMID- 12910642 TI - [Automatic erythrocytometry in a robotized microscope MEKOS-Ts1]. AB - Measurements of the form and size of erythrocytes are needed in the diagnosis of a number of diseases. However, such measurements, if made manually, are a labor consuming and often inaccurate method, which ensures the determination of a very limited number of parameters. Hardware/software unit MEKOS-C1 enables an automated examination of blood smear, thus speeding up significantly the analysis and ensuring a more complete and accurate information. The possibilities of unit MEKOS-C1 were evaluated for the diagnosis of ovalocytosis. Blood smears of 19 patients from the Russian Pediatric Clinical Hospital, including 8 patients with inherited ovalocytosis, 1 patient with spherocytic ovalocytosis and 10 patients without the disease, were made use of Erythrocytes were isolated in the images of preparations and the contour of each cell was approximated by ellipse. The ratio between ellipse semi-axes served as a measure of erythrocytes' ovality. The mean ratio of semi-axes (RS) and the index of ovalocytosis (IO), i.e. a ratio of the mean maximum diameter to the mean minimal diameter, were calculated for each smear. Manual IO measurements were made in all preparations as a control. Since an additional error can enter the result because of the irregular smear nature and impossibility to standardize completely the technology of smear preparation, the data, obtained from two different smear parts and from parallel preparations, were compared. The reliability, stability and good reproducibility of the automated measurement results were demonstrated. The mean erythrocytes' RS correlated well with IO, obtained manually, and did not virtually differ from RS, measured in the automated manner. The mean RS value of erythrocytes, obtained from patients with inherited ovalocytosis, significantly differed from the control values, which is indicative of a high information density of the discussed parameter. Therefore, RS, when measured automatically, is a reliable and convenient characteristic of erythrocytes' ovality under the conditions of using the ordinary technique of smear preparation. PMID- 12910644 TI - [Evaluation of results of total urine analysis in patients after kidney allotransplantation (review)]. PMID- 12910643 TI - [Clinical significance of determining the soluble transferrin receptor]. AB - The aim of the study was to work out a method for determining the soluble transferrin receptor (TfR) on the basis of the direct immune-enzyme analysis, including its dynamic testing in patients with anemia of various etiologies. Reagents, needed for the above analysis, i.e. TfRs and antibodies to them, were obtained to implement the set goal. TfR was isolated by Kanevsky's affine chromatography from the homogenate of patients. The thus isolated TfR was used to immunize and to obtain the monoclonal antibodies. The conjugate of horseradish peroxidase (HP) and antibodies to TfR were made use of to determine the quantity of bound antibodies. One type of antibodies were immobilized in plates of the "Nunk" company (Denmark), and the other type of monoclonal antibodies were conjugated with HP. A reliably higher TfR in iron-deficiency anemia was shown during the determination of the TfR level in 36 donors and 266 patients. After a conducted ferrotherapy, the TfR level went down approaching the normal value. The TfR level was related with a disease stage and activity of the prolipherative process in cases of autoimmune hemolytic anemia, B12-folate-dependent anemia, lympho- and myeloprolipherative diseases and in oncology patients. The elaborated method opens up new possibilities for the differential diagnosis of anemia and provide for objective criteria of a conducted therapy. PMID- 12910645 TI - [Use of test-system Amplicor HIV-1 for HIV-infection diagnosis in newborn in Russia--first results (2001-2002)]. PMID- 12910646 TI - [Tropical Medicine: towards scientific bilingualism without renouncing French speaking pedagogy]. PMID- 12910647 TI - [Mycetoma of the hand]. PMID- 12910648 TI - [Iron chelation in antimalarial therapy]. AB - Rapid development of significant resistance to antimalarial drugs has been a major force driving research to identify and develop new compounds. A number of iron(III)-chelating compounds designed for purposes other than treating malaria have in vitro antimalarial activity stemming from iron deprivation or toxic effects related to free radical release. Several of the iron(III) chelators have been effective in animal models of plasmodial infection. Desferrioxamine has been used successfully against both uncomplicated and severe malaria in humans. Iron chelating agents seem to be promising therapeutic adjuvants for treatment of severe Plasmodium falciparum malaria infection. PMID- 12910649 TI - [Practice guidelines for diagnosing the cause of hypereosinophilia in a traveler]. PMID- 12910650 TI - [Traveler's diarrhea: recommendations prior to departure and practice guidelines]. PMID- 12910651 TI - [Comparison of sociocultural attitudes towards epilepsy in Limousin (France), in Togo and in Benin (Africa)]. AB - Sociocultural attitudes continue to have a negative impact on management of epilepsy in many African countries and in a few advanced countries. The purpose of this study was to compare attitudes toward epilepsy in France and two African nations: Togo and Benin. A total of 305 epileptic patients over 18 years of age were interviewed using the same quantitative questionnaire about their beliefs, knowledge attitudes and practices regarding their disease. There were 77 patients from the Limousin region in France, 129 from the rural canton of Nadoba in Togo and 99 from the coastal province in Benin. The frequency of epileptic seizure during the last two years prior to the study was lower in France than in Togo and Benin. The number of people who believed in supernatural causes of epilepsy was higher in Togo and Benin whereas the number of people attributing the disease to social causes (e.g. death and stress) was higher in France. Few epileptic patients in France thought that the disease was contagious whereas many patients in both Togo and Benin still believed that the disease was contagious and that some foods were forbidden. More patients in France than in Togo and Benin were aware of the relationship of epilepsy with alcohol, drug abuse and cerebral injury. Epileptic patients in France were more likely to consult a physician and use medical drugs for the treatment of epilepsy than their counterparts in Togo and Benin. Epileptic patients in Togo often complained of social exclusion. Although sometimes contradictory, these findings support the idea that sociocultural attitudes should be taken into account in the management of the disease. PMID- 12910652 TI - [Post-hemolytic renal failure in children with glucose-6-phosphate dehydrogenase deficiency at the University Hospital Center in Lome]. AB - The purpose of the study was to identify predisposing factors for acute hemolysis and post-hemolytic renal failure in children with glucose-6-phosphate dehydrogenase deficiency (G6PD). Any child presenting hemoglobinuria during the study period was prospectively evaluated. Evaluation included detection of the presence of hemolytic agents, laboratory tests to measure hemolysis, G6PD activity, infection and renal failure, and assessment of outcome and management of hemolysis and renal failure. G6PD deficiency was documented in 32.1% of the 230 children admitted with hemoglobinuria. Anuric renal failure occurred during the hemolysis episode in 35.1% of patients with G6PD deficiency (21 boys and 5 girls between 30 months to 13 years old). Acute hemolysis associated with infection occurred before any treatment in 53.8% of cases and after beginning treatment in 46.1%. In 84.6% of cases, occurrence of acute hemolysis involved association of drugs considered as nonhemolytic either with themselves or with other drugs. Anuric renal failure occurred after beginning treatment in all cases and was most severe in patients with of multiple-germ infection (30.7%) and drug association (84.6%). Renal failure was reversible in 80.7% and fatal in 19.2%. Multiple-germ infection and drug association appeared as the main predisposing factors for post hemolytic anuric renal failure in patients with G6PD deficiency. The high frequency of these factors in tropical areas suggests implication of local endemic infections. PMID- 12910653 TI - [Seroprevalence of HIV infection in sexually abused children in Cameroon]. AB - The purpose of this study was to determine the prevalence of HIV infection in sexually abused children and again alert public official to the urgent need of a national committee on combating child sexual exploitation in Cameroon. This study was conducted at the New Bell District Hospital and Laquintine Hospital in Douala from February 1, 1998 to March 31, 2000. A total of 76 sexually abused children with a mean age of 11.6 years (range, 3-15) were included. There were 64 girls (84.2%) and 12 boys (15.8%). Among the 71 victims tested, 24 (37.5%) became seropositive as a result of sexual abuse with penetration. Abuse involved rape in 84.2% of cases and molestation in 15.8%. Abuse occurred within the family in 13.1% of cases and outside the family in 86.9%. Slightly more than 50% of the children studied did not attend school and 17.1% had menial jobs. This study demonstrates that sexual abuse is a major risk for sexually transmitted diseases including HIV. These findings also underline the need to adopt a national program to protect children from sexual abuse and to implement a policy for post-exposure prophylaxis against HIV. PMID- 12910654 TI - [Campaign to eradicate yaws on Santo Island, Vanuatu in 2001]. AB - At the beginning of the 1960s a campaign was carried out to eradicate endemic trepanematosis in the South Pacific. During the year 2000, twenty cases of yaws were diagnosed on Santo Island in Vanuatu. Recrudescence of this disease was confirmed by a seroprevalence testing showing 20.9% positive VDRL in 273 serum tests. In view of these findings, the Health Minister in collaboration with the WHO office in Vanuatu undertook mass benzathine-benzylpenicilline treatment in the population of the Santo and Malo (SANMA) province. The objective was to treat at least 85% of the population. Treatment was administered to 36,414 of the 39,397 inhabitants identified in 587 villages during the campaign, i.e., 92.4%. Coverage varied from 87.6 to 97.3% depending on the zone. Positive VDRL was obtained in 96 of the 230 serum samples collected for suspicion of yaws. All documented cases of yaws were found in the southeastern part of the island. Treatment had a dramatic effect on lesions within 24 hours. Cooperation of the population and involvement of mobile units and primary health-care providers were key factors in the success of this campaign. Further action will include training of health care workers and improvement of sanitary conditions in villages. Seroprevalence testing to assess the effectiveness of this campaign is to be carried out in 2003. PMID- 12910655 TI - [Paragonimiasis in Cameroon: clinicoradiologic features and treatment outcome]. AB - A prospective study of paragonimiasis was undertaken from September 1992 to July 1997 in the Mount Kupe zone of the Southwest Province of Cameroon. In a group of 2700 subjects, 312 presenting one or more signs of paragonimiasis underwent testing to detect Paragonimus africanus eggs in sputum and stools. Eggs were found in 30 subjects (9.61%). These patients underwent chest x-rays to assess radiological lesions due to paragonimiasis before and after treatment with Praziquantel. Symptoms included cough in all patients, hemoptysis in 13 (43.33%) and chest pain in 18 (60%). Only one patients presented altered general status. No patients presented fever. Before treatment chest x-rays demonstrated perinodular shadow in 22 patients (73.33%), pulmonary infiltrative opacity in 12 (40.00%), pulmonary nodules and cavitation in 3 (10.00%), pleuropulmonary calcification in 3 (10.00%), and cicatricial lesions in 2 (6.66%). Radiological findings were normal in 8 patients (26.66%). Following treatment parasitological and clinical cure was achieved at 1 and 2 months in all cases but X-ray abnormalities persisted for six months in 56.6% of cases. The most notable changes included disappearance of perihilar shadow in 8 out of 22 patients (36.36%) and worsening of radiological findings in 3 (13.66%). In 19 patients, treatment had no effect on X-ray findings in comparison with baseline. Absence of pleural effusion and high incidence of perihilar shadow may be specific features of paragonimiasis in Central Africa where the incidence of concomitant parasitic, fungal and microbial diseases is high. PMID- 12910657 TI - [American histoplasmosis: a case with cutaneous presentation in Morocco]. AB - Histoplasmosis Histoplasma capsulatum is a common systemic mycosis in immunocompromised subjects. It has not been reported in Maghreb countries. The purpose of this report is to describe a case with cutaneous presentation involving a Moroccan woman undergoing long-term corticosteroid therapy for systemic inflammatory diseases (rheumatoid polyarthritis and scleroderma). In addition to the main pathophysiological features of the disease, the authors present the causal agent and clinical manifestations. They stress the importance of laboratory findings in the diagnosis of this fungal disease. PMID- 12910656 TI - [Chloroquine drug prophylaxis no longer prevents Plasmodium falciparum-induced fetal hypotrophy in Kinshasa]. AB - Control of malaria-related antenatal morbidity and prevention of fetal hypertrophy due to Plasmodium falciparum-induced chorionitis in endemic malaria depends mainly on weekly treatment with chloroquine insofar as that drug is still effective. The purpose of this study was to evaluate the efficacy of chloroquine drug prophylaxis in preventing fetal hypotrophy in the hyper-endemic environment of Kinshasa, Democratic Republic of Congo, where emergence of chloroquine resistant P. falciparum strains is a current threat. Data concerning the mother, fetus, and placenta were collected from 59 parturients at Saint Marc's Maternity Hospital in Kingasani, which is one of the major health institution in the Democratic Republic of Congo. The confounding effect of parity, age, and placental weight was controlled using general linear model. Our results showed that regular chloroquine treatment did not lower the risk of chorionitis (Odds ratio = 2.0; CI95% = 0.4-11.6; p = 0.226) or its impact on birth weight. Findings after correction showed that mean birth weight was significantly lower (p = 0.011) with (2.724 +/- 89 g; 24 cases) than without (2.992 +/- 80 g; 32 cases) occurrence of chorionitis. PMID- 12910659 TI - ["Epidemic" of plantar dermatitis during military maneuvers in the rain forest of Guyana]. AB - The purpose of this report is to describe an "epidemic" outbreak of plantar dermatitis that occurred in 30 soldiers on mission in the rain forest of Guyana. The contingent was forced to stop twice and operational readiness was seriously reduced. The first episode affecting 43% (13/30) of the contingent occurred after 4 days of march. In the second episode 70% (21/30) of the contingent was affected. Manifestations were characterized by the appearance of distinct, dry, nonpruriginus, erythematous papules followed a few hours later by quasi exfoliating, noncoalescing, nonoozing, nonmalodorous lesions measuring several centimeters in diameter. Symptoms regressed quickly within 48 to 72 hours with only symptomatic treatment. A combination of microtrauma and maceration is the most likely mechanism. However identification of an offending microbial, viral or fungal agent or of a cause in the environment, clothing or products could not be made. A tempting hypothesis involves an inflammatory reaction caused by an organism in ground water or the soil. A prospective study could be undertaken at the Rain Forest Training Camp in Regina where similar clinical manifestation have been observed. PMID- 12910658 TI - [Heterotopic pregnancy: report of three cases]. AB - The association of intra-uterine and extra-uterine pregnancy has increased steadily with the widespread use of infertility treatment. Diagnosis of extra uterine pregnancy may be delayed as a result of visualization of an intrauterine gestation sac, leading to a less favorable diagnosis than for simple extrauterine pregnancy. The purpose of this report is to described three heterotopic pregnancies observed in a series of 25 simple extrauterine pregnancies managed in the Gynecology Department of the El Rapha Clinic in Libreville Gabon over an 18 months period. A review of the literature is included to provide more background on this association. PMID- 12910661 TI - [Chronic tropical pancreatitis: a case report]. AB - The purpose of this report is to describe a case of tropical calcific pancreatitis (TCP). This disease is specific to tropical regions and constitutes the main cause of chronic pancreatitis in children worldwide. It can also be observed in young adults (2nd and 3rd decade). Shortage of dietary lipids during childhood has been implicated in the development of TCP and mutation of the SPINK1 gene has been cited as a predisposing genetic factor. The underlying pathophysiology of TCP is the same as chronic calcific pancreatitis (CCP) due to alcohol abuse. The main features are a sex ratio of 1, absence of alcohol consumption, occurrence of childhood diabetes in one third of cases, low incidence of acidoketosis, and presence of macro-calcifications especially in ducts. In 10% of cases TCP is complicated by pancreatic carcinoma occurring at an early age, located mainly in the body and tail of the pancreas, and having a less favorable prognosis than primary cancer. Treatment of patients with TCP is the same as for patients with CCP due to alcohol abuse. Prevention depends on improvement of nutritional status of the population. PMID- 12910660 TI - [Sporadic porphyria cutanea tarda: a case report in a Moroccan man]. AB - This report describes a case of sporadic porphyria cutanea tarda involving a 38 year-old Moroccan man. Clinical diagnosis was based on characteristic features, i.e., facial hypertrichosis and bullous lesions lasting four months during the summer of 2000 followed by macular scarring on the dorsal surfaces on the hands. Three well-known precipitating factors were noted, i.e., sun, ethanol and hepatitis C virus infection. Laboratory diagnosis was based on dark red urine and elevated serum and urine uroporphyrin levels. Enhanced uroporphyrin production was due to urodecarboxylase deficiency in the liver. Urodecarboxylase activity in red blood cells and serum ferritin level were normal. The patient is heterozygous for the His63Asp HFE gene mutation associated with hereditary hemochromatosis. The photoprotective effect of melanin in this dark-skinned patient failed to offset uroporphyrin-induced photosensitivity. Avoidance of sun, ethanol and phlebotomy have prevented recurrences. PMID- 12910662 TI - [Description of the first cases of serotype A, sequence type (ST)-11 meningococcal meningitis in Senegal]. AB - Senegal is located in the African meningitis belt and meningococcal meniningitis outbreaks are yearly events. Occurrence of an epidemic involving serogroup W135 in 2000 and its spread following the Hajj (pilgrimage to Mecca) exposed the strongly Moslem population of Senegal to the risk of early infection. Indeed the first two cases in Dakar occurred simultaneously with the spread of this epidemic strain. The purpose of this article is to describe clinical, laboratory, and therapeutic findings in these two cases and the results of the ensuing epidemiological survey. The relationship with the pilgrimage and consequences on public health in Senegal are discussed. PMID- 12910663 TI - [Outbreak of tropical phagedenic ulcers after the rainy season in Dijibouti]. AB - The incidence of cutaneous ulcers was observed after the rainy season in Djibouti in 1997. Based on the study of epidemiologic, clinical, biological, and therapeutic features these lesion were classified as phagedenic ulcers. While direct examination showed numerous fusiform bacilli, cultures performed in one patient, led to isolation of numerous colonies of Prevotella loescheii. Though less common, two other anaerobic bacterial species were detected, i.e., Peptostreptococcus anaerobius and Peptostreptococcus sp. PMID- 12910664 TI - [Chronic malnutrition in adolescents from 10 to 19 years of age living in rural Togo (from August to October, 1997 and November to December, 1998)]. PMID- 12910665 TI - [Trichinosis in Papua New Guinea]. PMID- 12910666 TI - [...PAL+ in Anglet]. PMID- 12910667 TI - [In vitro differentiation of rat bone mesenchymal stem cells into neuron-like cells induced by vitamin A acid etc]. AB - OBJECTIVE: To explore the feasibility of inducing rat bone mesenchymal stem cells (BMSC) to differentiate into neuron-like cells with the use of Vitamin A acid, zinc and rat injured spinal cord extracts in vitro. METHODS: The BMSC were isolated from rat, cultured for 4 passages, and were treated with 10 ng/ml basic fibroblast growth factor (bFGF) for 24 h before induction. Then the medium was replaced by an induction media containing Vitamin A acid, zinc and rat injured spinal cord extracts. The morphological changes of the cells were observed. At day 12 of induction, the cells were stained immunocytochemically with neuron specific enolase (NSE), neurofilament (NF) and glial fibrillary acidic protein (GFAP) antibodies. RESULTS: At day 12 of induction, a certain number of BMSC became neuron-like cells and showed NSE and NF expression. But the neuron-like cells did not express GFAP. CONCLUSION: The BMSC can be induced to differentiate into neuron-like cells with the use of Vitamin A acid, zinc and rat injured spinal cord extracts. PMID- 12910668 TI - [Adipose tissue-derived stromal cells differentiate into neuron-like cells]. AB - OBJECTIVE: To investigate the possibility of inducing adipose tissue-derived stromal cells to differentiate into neuron-like cells, and to explore a new cell source for central nervous system transplantation. METHODS: beta-mercaptoethanol was adopted to induce the cells to differentiate; undifferentiated cells and differentiated cells were identified with immunocytochemistry. RESULTS: A population of adipose tissue-derived stromal cells were isolated from adult rat adipose tissue; they were processed to obtain a fibroblast-like population of cells and could be maintained in vitro for extended periods with stable population doubling, and they were expanded as undifferentiated cells in culture for more than 10 passages, indicating their proliferative capacity. beta mercaptoethanol induced the stem cells to express nestin, characteristic of neuronal precursor stem cells at early stage of differentiation, and at late stage they exhibited a neuronal phenotype, expressing neuron-specific enolase (NSE) and neurofilament(NF); with an optimal differentiation protocol, almost 60% 85% of the cells expressed NSE and NF. CONCLUSION: The data support the hypothesis that adult adipose tissue contains stem cells capable of differentiating into neurons. PMID- 12910669 TI - [Construction of plant expressive vector of human beta-defensin-2 gene]. AB - OBJECTIVE: Several evidences suggested that transgenic plants would be a facile and economic bioreactor for large-scale production of industrial and pharmaceutical recombinant proteins. This study is made in an attempt to establish plant bioreactor for expression of recombinant hBD-2. METHODS: Recombinant hBD-2 gene with C terminal of bi-tags of myc and 6xHis was inserted into a plant expressive vector-pCAMBIA1304, which closely located the down-stream of CaMV35S promoter. Agrobacterium tumefaciens LBA4404 was transformed with the recombinant plant expressive vector: rpCAMBIA1304/hBD-2/His. The positive clones of LBA4404 transformed by rpCAMBIA1304/hBD-2/His were selected on a culture plate containing kanamycin. The callus tissues were transfected by positive clones of LBA4404, and positive callus were examined by using the resistant selection of hygromycin gene. RESULTS: The evidences of enzyme digestion, PCR and sequence analysis confirmed that recombinant hBD-2 gene with C terminal of bi-tags of myc and 6xHis was correctly inserted into pCAMBIA1304 and was located between CaMV35S promoter and Nos terminal cordon to construct recombinant plant expressive vector: rpCAMBIA1304/hBD-2/His, thus indicating that rpCAMBIA1304/hBD-2/His has successfully transformed Agrobacterium tumefaciens LBA4404 and positive clones have been isolated. The results from resistant selection of hygromycin gene showed that rpCAMBIA1304/hBD-2/His has been transferred into the callus of wheat, and the differentiation of callus tissue under selective pressure of hygromycin is carried out continually. CONCLUSION: The above data suggest that the technique of transgenic plant is workable for the production of recombinant hBD-2. PMID- 12910670 TI - [Construction of a site-directed mutant of FALL-39 and antibacterial activity of its E. coli-based product]. AB - OBJECTIVE: Constructing a point-directed mutant of FALL-39 and prokaryotic expressive vector, and determining its antibacterial activity. METHODS: A two step polymerase chain reaction (PCR) was used for the site-directed mutagenesis. Two sets of primers (P1, P2, P3, P4) were designed according to FALL-39 gene sequence and mismatch was introduced into P2 for a substitution of AAG for AAT at position 32. Mutagenesis was performed in a two-step PCR, the amplified fragments from the second PCR, which contain the mutation site, were subcloned into the vector PGEX lambda 1T and verified by sequencing analysis. Antibacterial activities of the E. coli-based product against E. coli and P. aeruginosa were detected by using minimal inhibitory concentration (MIC), minimal effective concentration (MEC) and minimal bactericidal concentration (MBC). RESULTS: A mutant of FALL-39, whose AAT at position 32 is substituted by AAG, was obtained by the two-step PCR. The antibacterial activities of the E. coli-based product of the recombinant FALL-39-Lys32 against E. coli ML-35p and P. aeruginosa ATCC27853 were much stronger than those of FALL-39. MIC and MBC of the FALL-39 against E. coli were 18.75 micrograms/ml and 37.50 micrograms/ml, whereas those of FALL-39 Lys32, 10.94 micrograms/ml and 21.88 micrograms/ml respectively. CONCLUSION: A point-directed FALL-39-Lys32 mutant was obtained by using two-step PCR and its antibacterial activity was increased, suggesting that increased cationic FALL-39 mutant might enhance its antibacterial activity. PMID- 12910671 TI - [Multiple human cord blood transfusion for inducing low immune response]. AB - OBJECTIVE: To investigate and evaluate a clinically perspective way of inducing transplantation tolerance. METHODS: Multi-cord blood transfusion was performed before kidney transplantation in 20 cases. Donor-specific mixed lymphocyte reaction(MLR) and allograft rejection were observed and studied. RESULTS: The donor-specific MLR was suppressed in every recipient. No allograft rejection took place in the cord blood transfusion group three months after kidney transplantation. CONCLUSION: Multi-cord blood transfusion is a practical way of inducing transplantation tolerance. PMID- 12910672 TI - [Research on expansion of CD34+ cells from umbilical cord blood using different cytokine groups in vitro]. AB - OBJECTIVE: To investigate the expansion of the CD34+ cells of umbilical cord blood in vitro culture comprising different cytokines and the potential hematopoiesis function of the expanded CD34+ cells. METHOD: Expanding the purified CD34+ cells from the fresh umbilical cord blood for 6 days in vitro with the use of different combination of FL, SCF, TPO, IL-3 and IL-6. Getting one better combination by analyses of the results from flowcytometry, LTC-IC, and progenitor clonogenic assays. RESULTS: The numbers of CD34+ cells expanded by use of different combination of cytokines were 3.1-8.6 times as large as the numbers of ordinary ones, thus making 16 cases of insufficient CD34+ cells become able to satisfy the needs of the adults weighing 50 kg. No significant difference was observed between the function of the expanded cells and the original ones in hematopoiesis. Meanwhile, it was proved that TPO in combination with other cytokines could strengthen the expansion of CD34+ cells, but using TPO alone for long time culture would cause more CD34+ cells to be megalocaryocytes. CONCLUSION: The combination of FST36 cytokines can expand the CD34+ cells of the umbilical cord blood to meet the adults' needs in laboratory's methods. PMID- 12910674 TI - [Plasma haemostatic and fibrinolytic activities and their relationship to levels of serum lipids and apolipoproteins in endogenous hypertriglyceridemic patients]. AB - OBJECTIVE: To investigate the changes of plasma haemostatic and fibrinolytic activities and their relationship to serum lipids and apolipoproteins levels in endogenous hypertriglyceridemic patients. METHODS: We determined the plasma prothrombin time(PT), activated partial thromplastin time(APTT), activity of tissue-type plasminogen activator(t-PA), activity of plasminogen activator inhibitor-1 (PAI-1) and fibrinogen(Fg) level of 24 endogenous hyper triglyceridemic patients and 34 healthy controls, and observed their relationship to serum lipids and apolipoproteins. RESULTS: The PT and APTT of patients with hypertriglyceridemia were obviously shorter than those of control subjects (P < 0.01, P < 0.05); PAI-1 of patients was significant higher than that of control group (P < 0.01); there was no statistically significant difference in t-PAI-1: a between the control subjects and the hypertriglyceridemic patients; the levels of Fg in patients was 27% higher than that in healthy control (P < 0.01). The correlation analysis indicated that PT was significantly associated with the levels of plasma HDL-C (r = 0.445, P < 0.01), and inversely associated with the levels of plasma TG, apoC III, BG and BMI (r = -0.294, -0.320, -0.282, -0.272, P < 0.05); APTT was negatively correlated with serum TG and apoC III levels (r = 0.345, -0.320, P < 0.05); t-PA was negatively correlated with BG levels (r = 0.336, P < 0.01). PAI-1 and Fg levels were strongly correlated with serum TG, apoC II, C III and E levels (r = 0.400, 0.408, 0.497, 0.454, P < 0.01 and r = 0.642, 0.581, 0.673, 0.304, P < 0.01, respectively), and negatively correlated with HDL-C (r = -0.366, -0.524, P < 0.01). CONCLUSION: Increasing of coagulating activity and decreasing of fibrinolytic activity in hypertriglyceridemic patients were significantly associated with serum lipids and apolipoproteins levels. PMID- 12910673 TI - [Screening of hypercoagulability-related genes in aorta of hypertriglyceridemic rats]. AB - OBJECTIVE: The aim of this study was to screen hypercoagulability-related genes in aorta of hypertriglyceridemic rats. METHODS: Suppression subtractive hybridization (SSH) was used to construct hypercoagulable rat cDNA subtractive library. Semi-quantitative RT-PCR was applied to identify differentially expressed genes. RESULTS: Hypercoagulable rat cDNA subtractive library had been successfully constructed. Sequence analysis and homology searching of 20 clones showed that these clones represent 19 genes: 8 are known genes newly found to have a relationship to hypercoagulability and 11 are novel genes (ESTs) that had been deposited to GenBank. Three ESTs (HC002, HC057 and HC067) were confirmed to be over-expressed genes by semi-quantitative RT-PCR. CONCLUSION: The constructed subtractive library is highly efficient and has laid a foundation for screening and cloning hypercoagulability-related novel genes of rat, HC002, HC057 and HC067 are probably related to hypercoagulability induced by hypertriglyceridemia. PMID- 12910675 TI - [Apoptosis of cancer cells induced by influenza virus and its elementary mechanism]. AB - OBJECTIVE: To study the apotosis-inducing effects of influenza virus on cancer cells and the role of FasL. METHODS: Four strains of cancer cells (Hela, Raji, SMMC-7721 and SPC-A-1) were infected with 20 m. o. i influenza virus and the effects of apoptosis were detected at different time points post infection by electron microscopy, DNA agrose gel electrophoresis, PI stained flow cytometry, and Annexin-V FITC/PI stained flow cytometry. And the concentration of FasL in the culture supernatant was measured with the use of ELISA at the same time. RESULTS: All the four strains of cancer cells showed apoptotic morphological changes. DNA agarose electrophoresis also showed "ladder" in a time dependent manner after virus infection. During this course, there was an obvious increase of FasL concentration in the cell culture supernatant. CONCLUSION: Influenza virus can induce tumour cells apoptosis in vitro, and it is probably related to the increased expression of FasL. PMID- 12910676 TI - [Study on the antisense inhibition effect of liposome-mediated radiolabled antisense oligonucleotides of c-myc mRNA]. AB - OBJECTIVE: To ascertain whether liposome-mediated antisense oligonucleotides could inhibit the cancer cells from growing and the oncogene from expressing. METHODS: The antisense, sense and scrambled oligonucleotides for c-myc mRNA, each containing 15 bases, were synthesized elsewhere and were labeled with 99m technetium (99mTc-DNA). Some of them were encapsulated with liposome. Then the 99mTc-DNA and the 99mTc-DNA entrapped with liposome were transfected into colon cancer cells, LS-174-T, with different radioactivity concentrations. At different time after transfection the uptake rate of cells was measured, and the reflow rate was detected at 18 h. Inhibition effect on cellular growth was tested by MTT colorimetric experiment and on the expression of c-myc by flow cytometry. RESULTS: The cellular uptake rate increased gradually after transfection within 5 hours, and that of liposome-mediated 99mTc-DNA was significantly higher than that of 99mTc-DNA. The reflow rate and fluorescence intensity of oncoprotein were 39.51 percent and 2.9860 +/- 0.3733 for antisense oligonucleotides, 44.12 percent and 4.2600 +/- 0.2218 for sense oligonucleotides, and 63.92 percent and 5.2620 +/ 0.8562 for mismatched oligonucleotides. The reflow rate and fluorescence intensity of liposome-entrapped 99mTc-labeled antisense oligonucleotide were significantly lower than those of sense and scrambled oligonucleotides. CONCLUSION: The liposome-mediated 99mTc-labeled antisense oligonucleotides could prohibit the cancer cells from growing and the oncogene from expressing. PMID- 12910677 TI - [The changes of angiotensin II receptor expression in cardiomyocytes of ventricles in pigs with experimental ventricular septal defect]. AB - OBJECTIVE: To investigate the changes of angiotensin II receptors expression in cardiomyocytes of both ventricles in pigs with experimental ventricular septal defect (VSD). METHODS: Nineteen young pigs were divided into 3 groups: the pigs that received operation to produce experimental VSD, the pigs that underwent sham operation, and the pigs as normal control. One month after the operation, the pigs were catheterized and then put to death; the cardiomyocytes were taken from their ventricles and were isolated. Radioligand binding assay was done to measure the Bmax and KD of angiotensin II receptor per 10(6) cells. RESULTS: For both Bmax and KD of angiotensin II receptors, there were no significant differences between the sham-operation group and the control. The Bmax of left ventricle AT1R (188.42 +/- 133.97 fmol/10(6)) and Bmax of right ventricle AT1R (272.14 +/- 232.74 fmol/10(6)) and AT2R (40.42 +/- 34.76 fmol/10(6)) in the operation group were significantly elevated in comparison with the control (Bmax of left ventricle AT1R 29.20 +/- 19.50 fmol/L, and Bmax of right ventricle AT1R 76.72 +/- 51.21 fmol/L and AT2R 9.63 +/- 1.27 fmol/L) respectively, P < 0.05. CONCLUSION: Intracardiac left-to-right shunt related volume overload leads to the elevation of AT1R expression in cardiomyocytes, which serves as an important mechanism of ventriclular hypertrophy and remodling in congenital heart disease. The increased expression of AT2R in right ventricle is a possible explanation of the phenomenon that moderately sized VSDs usually show left other than right ventriclular hypertrophy, taking into account that AT2R has been suggested to mediate antitrophic reaction. PMID- 12910678 TI - [Optimizing cultural conditions of measles vaccine working seed lot with orthogonal experiments]. AB - OBJECTIVE: To improve the quality of measles vaccine and find out good combination of cultural factors of measles vaccine virus seeds. METHODS: Orthogonal experiments were made to confirm optimized cultural factors. 5 lots of optimized measles vaccine working seed lot were prepared. RESULTS: The titers of the optimized vaccine seed improved significantly (> or = 0.69 +/- 0.35 LgCCID50/1.0 ml, P < 0.05), compared with 5 lots of traditional working seed lot. With these seeds we trial-prepared 5 lots of lyophilized vaccines respectively, the titer and stability met the Chinese Requirements of Biological Products. CONCLUSION: The orthogonal experiment is a kind of good method for optimizing cultural conditions of measles vaccine working seed lot. The yield and quality of measles vaccine can be improved in the optimized working seed lot with high titer and good stability. PMID- 12910679 TI - [The expression of A103 and inhibin alpha in adrenocortical adenoma by high throughput tissue microarray techniques]. AB - OBJECTIVE: To evaluate the potential diagnostic role of A103 and inhibin alpha in adrenocortical adenoma and the feasibility of tissue microarray/tissue chips. METHODS: A series of adrenal gland tissue chips were prepared, including 79 cases of formalin-fixed, paraffin-embedded tissue from 3 normal adrenal glands, 66 adrenocortical adenomas, 5 cases of metastatic carcinoma and 5 pheochromocytomas. A103 and inhibin alpha were detected with Envision immunohistochemical staining methods. RESULTS: Positive staining was seen in 3 of 3 (100%) adrenal cortices and in 55 of 61(90.2%) adrenocortical adenomas but in none of the pheochromocytomas or adrenal metastatic carcinomas. In the 3 of 3(100%) adrenal cortices, inhibin alpha immunoreactivity was limited to the zona reticularis and the innermost zona fasciculata. 51 of 61(83.6%) adrenocortical adenomas reacted with inhibin alpha. None of the pheochromocytomas or metastatic carcinomas reacted with inhibin alpha. CONCLUSIONS: High-throughput tissue microarray/tissue chip offers a powerful strategy for rapid identification in situ. It is feasible to utilize tissue microarray for a rapid, economic and accurate screening of tissue specimens on a large scale. The combined use of A103 and inhibin alpha is valuable in distinguishing adrenocortical adenoma from pheochromocytoma and other metastatic carcinomas. PMID- 12910680 TI - [Polymorphism of insulin receptor substrate-1 gene in patients with coronary heart diseases]. AB - OBJECTIVE: To explore the relationship between the polymorphism of insulin receptor substrate-1 (IRS-1) gene and coronary heart diseases in Chinese population. METHODS: A total of 113 patients with coronary heart diseases and 194 healthy Chengdu residents with less than 82 mmol/L of TG and 6.2 mmol/L of TC in fasting serum testing were selected and compared. The genotype and allele frequency of IRS-1 gene polymorphism was assayed by polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP). The serum lipids were measured by enzymatic kits. RESULTS: The GG genotype dominated in the IRS-1 gene with frequency of 0.960 and 0.985 for patients and healthy controls respectively. Difference appeared in R allele frequency (0.015 for healthy controls and 0.040 for patients, P = 0.05). Patients with genotype G/R had significantly higher level of serum TC and lower level of plasma glucose than patients with genotype G/G (P < 0.05). CONCLUSION: IRS-1 gene polymorphism is associated with coronary heart disease in Chinese population. The G/R and G/G genotype of IRS-1 gene contributes to the increasing of serum cholesterol and plasma glucose respectively. PMID- 12910681 TI - [Internalization of antibody-targeted immunonanoparticles into human hepatoma cells and its reversal effect on MDR]. AB - OBJECTIVE: To observe if antibody-targeted immunonanoparticles could internalize into sensitive and multidrug resistance (MDR) cells of human hepatoma; and to study if the immunonanoparticles could reverse the MDR. METHODS: Human hepatoma specific adriamycin-loaded human serum albumin immunonanoparticles (HAb18-ADR-HSA NP) were incubated with human hepatoma sensitive cell line (SMMC-7721) or MDR cell line (SMMC-7721/MDR+) and the internalization of immunonanoparticles were observed by laser confocus microscopy, scanning electron microscopy and transmission electron microscopy; MTT colorimetric assay was used for assaying in vitro cytotoxicities of HAb18-ADR-HSA-NP to the resistant variant cells. Then based on these data, IC50 value of the immunonanoparticles and RF (Resistant Factor) of MDR cells were calculated. RESULTS: Laser confocus microscopy showed that many fluorescent particles (labeled immunonanoparticles) tightly adsorbed to SMMC-7721 cells and were also seen in cytoplasm of SMMC-7721 cells. When incubated with immunonanoparticles at 37 degrees C, many of immunonanoparticles were visualized in cytoplasm of SMCC-7721 or SMCC-7721/MDR+. These immunonanoparticles-contained cells exhibited damaged ultrastructures and the damage degree depended on incubation time. When the human hepatoma cells were pretreated with HAb18 antibody and incubated with immunonanoparticles, few immunonanoparticles were seen in cytoplasm of the cells, suggesting antibody specific internalization of the immunonanoparticles. Scanning electron microscopy demonstrated specific binding of the immunonanoparticles to the resistant variant cells. That immunonanoparticles exerted enhanced cytotoxicity to the resistant variant cells was demonstrated by a decrease of RF value of MDR cells, compared with free ADR (4.4 vs. 2.1). CONCLUSION: Human hepatoma-specific adriamycin loaded human serum albumin immunonanoparticles could specifically internalize into sensitive or multidrug resistance cells of human hepatoma via antibody direction. The immunonanoparticles could enhance the sensitivity of MDR cells to ADR cytotoxicity, suggesting its reverse effect on MDR. PMID- 12910682 TI - [Investigation on tumor-targeting characteristics of bifidobacterium infantis to melanoma in mice]. AB - OBJECTIVE: To investigate the tumor-targeting of bifidobacterium infantis to melanoma. METHODS: After bolus administration of bifidobacterium infantis with 3H TdR tagged, the values of radioactivity in tumor and organs were examined at days 1, 2, 3, 4 and 7. Anaerobic culture and histological observation of tumor and normal organs were taken for the examination of tumor-targeting characteristics of bifidobacterium infantis. RESULTS: The radioactivity in melanoma tissue increased progressively, while the radioactivity in normal organs became attenuated with time. The anaerobic culture showed an obvious proliferation of bifidobacterium infantis in tumor tissue. A large part of area was Gram positive in the tumor tissue section, whereas the normal tissue was Gram negative. CONCLUSION: Bifidobacterium infantis has good tumor-targeting characteristics in mice melanoma. PMID- 12910683 TI - [Effects of quercetin on the proliferation and apoptosis in transplantation tumor of breast cancer in nude mice]. AB - OBJECTIVE: To investigate the effects of quercetin on tumour growth, cell proliferation and apoptosis in transplantation tumor of breast cancer cell line MCF-7 in nude mice. METHODS: MCF-7 cells were inoculated into the mammary fatty pad of nude mice to establish breast cancer model, then twenty-four BALB/c nude mice with xenograft tumor were randomized into four groups: Control group, Quercetin group, 5-Fu group and Quercetin + 5-Fu group. After 15 days treatment, samples of tumor were collected. The sections of tumor were observed under light microscope and electron microscope. Cell apoptosis in situ was examined by a Tunel assay, and the expressions of ki67 antigen and B-cell lymphoma/leukemia-2 (Bcl-2) were detected by immunohistochemistry. RESULTS: 1. The tumor weight of Control group was significantly higher than those of Quercetin group, 5-Fu group and Quercetin + 5-Fu group (P < 0.05). 2. Immunohistochemical staining of ki67 showed that the ki67 label index (ki67-LI) displayed significant difference between Control group and Quercetin group, 5-Fu group, Quercetin + 5-Fu group, and so did the staining of Bcl-2. 3. Detection of apoptosis in situ showed that apoptosis index (AI) was significantly higher in Quercetin group, 5-Fu group and Quercetin + 5-Fu group than in control group (P < 0.01), and higher AI was observed in Quercetin + 5-Fu group as compared with the AI in Quercetin group and 5-Fu group (P < 0.05). CONCLUSION: Quercetin can inhibit the growth of transplantation tumor of breast cancer cell line MCF-7 in nude mice. PMID- 12910684 TI - [The function of integrin beta 1 in the hypoxic cellular damage mechanism of renal tubular epithelial cell of newborn porcine]. AB - OBJECTIVE: To inquire into the mechanism of renal deficiency in neonatal asphyxia. METHODS: The 30, 60, 90, 120 min hypoxic renal cell model was made by use of DMEM/F-12 with antimycine A. ATP content was detected by reversed-phase HPLC. The levels of integrin beta 1 of renal tubular epithelial(RTE) cell of the groups were detected using flow cytometry. RESULTS: ATP deficiency cellular model of newborn porcine RTE cell was successfully set up. The levels of ATP of the newborn porcine RET cell during 30, 60, 90, 120 min hypoxia time significantly decreased as compared with normal control (P < 0.001). The levels of integrin beta 1 of newborn porcine RTE cell decreased as the ATP deficiency time increased and the differences between the groups were statistically significant (P < 0.001). CONCLUSION: The decreased level of integrin beta 1 in the newborn porcine RTE cell during anoxia may play an important role in the newborn renal deficiency during asphyxia. PMID- 12910685 TI - [The changes of filamin distribution in renal tubular epithelial cells during kidney ischemia/reperfusion injury in neonatal rats]. AB - OBJECTIVE: By investigating the changes of filamin distribution in renal tubular epithelial cells at different ischemia/reperfusion time stages we studied the role of filamin in kidney ischemia/reperfusion injury. METHODS: The neonatal SD rat ischemia/reperfusion model was set up. Immunofluorescent staining was used to show the distribution of filamin in renal tubular epithelial cells and the quantities of filamin were counted by ImagePlus-Pro system. RESULTS: Filamin was located around the basal plasma membrane in normal renal tubular epithelial cells. After ischemia for 0.5 h, the redistribution was not significant. After reperfusion for 0.5 h, filamin came into the cell plasma, appeared at the apex of cells and in the lumen. This kind of change was most apparent after reperfusion for 2 h and accompanied with destruction of renal tubules. The regeneration started 24 h post-ischemia. Filamin was redistributed to the basal plasma membrane. After reperfusion for 120 h, the regeneration ended and the structure of renal tubules became normal. CONCLUSION: The distribution of filamin was changed during ischemia/reperfusion injury, this change was prior to the change of actin cytoskeleton and integrin. PMID- 12910686 TI - [A study on the protective efficacy of yisheng injection against IRI in pig renal transplant model and the mechanism therein involved]. AB - OBJECTIVE: To inquire into the mechanism of the protective efficacy of Yisheng injection against ischemia/reperfusion injury(IRI) in pig renal-transplant model. METHODS: A total of 24 Chinese Sichuan White Pigs (SWPs) (2-3 months old, 17.5 +/ 4.6 kg, with clear family background) were used. 6 pairs of SWPs with better results of tissue typing were allocated to the group of renal allotransplantation. The other SWPs were subjected to renal autotransplantation. Yisheng injection was added into the preservative fluid and injected intravenously after transplantation in the test groups against IRI. We detected the SWPs' liver and kidney functions and serum superoxide dismutase(SOD) and malondialdehyde(MDA) concentrations after transplantation. RESULTS: Serum creatinine and urea levels increased in 2 days after operation and then returned to normal in both groups except the pigs suffering from acute tubular necrosis or acute rejection. The concentrations of serum MDA, SOD began to increase 2 days after operation (P < 0.05). Less increase of serum MDA, less decrease of NO and more increase of SOD in test groups were detected, compared with those in control groups (P < 0.05). CONCLUSION: Yisheng injection has protective efficacy against IRI and hence may have an application foreground. PMID- 12910687 TI - [Protective effect of berberine on cardiac myocyte injured by ischemia reperfusion]. AB - OBJECTIVE: To observe the protective effect of berberine on cardiac myocyte injured by ischemia-reperfusion(I/R) in neonatal rats. METHODS: Cardiac myocytes from neonatal SD rat were cultured and pretreated with berberine in three different concentrations, 1.5 x 10(-6) mol/L, 1.5 x 10(-5) mol/L and 1.5 x 10(-4) mol/L, before exposure to hypoxia (95% N2-5% CO2) for 24 h and reoxygenation (95% air-5% CO2) for 1 h to create cell model of ischemia-reperfusion. Then lactate dehydrogenase (LDH) release, methylenedioxyamphetamine (MDA) release superoxide dismutase (SOD) activity were measured, and cell apoptosis was detected. RESULTS: Compared with the measurements taken in normal group, the LDH and MDA in the supernatant of the cells in ischemia and reperfusion group increased highly (P < 0.01), SOD activity decreased sharply (P < 0.01), and apoptosis of cells in ischemia groups and reperfusion groups increased highly (P < 0.01). Pretreatment of myocytes with berberine resulted in reduction in LDH and MDA release (P < 0.01) in I/R groups, attenuation of apoptosis in ischemia and reperfusion groups (P < 0.01). When the concentration of berberine was increased, these effects were getting obvious. Especially when the myocytes were pretreated with berberine at 1.5 x 10(-5) mmol/L, the cell apoptosis rates were 14.4% and 20% in ischemia group and reperfusion group, which were lower than the rates of 17.4% and 41% before pretreatment, respectively. CONCLUSION: Berberine alleviates I/R injury and attenuates apoptosis in myocytes exposed to I/R. These effects partly depend on the concentration of berberine. PMID- 12910688 TI - [Cerebral ischemic tolerance induced by focal ischemic preconditioning in rats]. AB - OBJECTIVE: To investigate the ischemic tolerance induced by focal ischemic preconditioning (PC) in rats. METHODS: Ten minutes of middle cerebral artery occlusion(MCAO) was used for PC. Different duration of reperfusion (1, 3, 7, 14, 21 and 28 days) was allowed before 2 hours of MCAO followed by 22 hours of reperfusion in SD rats that were divided into the PC group and the PC plus MCAO group, and were compared with the rats of sham-operation plus MCAO group. Neurological scores, infarct volume, brain water content and HE staining were evaluated in each group. RESULTS: PC produced no neurological deficits and TTC demonstrated infarct. PC reduced neurological deficits significantly caused by 2 h MCAO 1-28 days after PC. Infarct volume was reduced only if PC was performed 1 14 days before MCAO. PC given 3 days before MCAO also decreased brain water content. CONCLUSION: Ten minutes of MCAO was strong enough to induce ischemic tolerance without brain injury. Infarct volume reduction appeared at 1 day and did not vanish until 14 days after PC, but protection against neurological impairment existed at least 4 weeks. PMID- 12910689 TI - [Study on the effect of integrin on human sperm activation]. AB - OBJECTIVE: This study was aimed to disclose whether the VLA-5 integrin on human sperm membrane takes part in the activation of human spermatozoa. METHODS: Fluo 3/AM was coincubated with human spermatozoa. The change of intracellular calcium([Ca2+]i) was determined by flow cytometry when human sperms were exposed to Fibronectin(Fn) as the specific ligand of integrin VLA-5. Human sperm acrosome reaction(AR) rate was evaluated by triple-stain technique. RESULTS: Fn significantly raised the AR rate (P < 0.05) and the level of [Ca2+]i (P < 0.01) in the spermatozoa of fertile men. But there was no significant rise in the level of [Ca2+]i in the spermatozoa of infertile men. CONCLUSION: The VLA-5 integrin on human sperm membrane plays a role in the activation of sperm. PMID- 12910690 TI - [Effect of removal of superior spermatic nerve on spermatogenesis]. AB - OBJECTIVE: To evaluate the role of superior spermatic nerve in the regulation of spermatogenesis. METHODS: Mature SD rats (350-375 g) received bilateral surgical removal of the superior spermatic nerve(SSN). The animals were killed 1 and 2 months after operation and were studied in comparison with the SD rats of the sham operation group. H. E. staining was employed in observing spermatogenesis, and transmission electron microscopy and terminal deoxynuleotidyl tansferase mediated dUTP nick end-labeling (TUNEL) were used to detect apoptosis. RESULTS: No pathological changes were observed 1 month after operation. Two months after SSN denervation, spermatogenesis arrest was seen in the seminiferous tubules in which only Sertoli cells or Sertoli cells and a few spermatogonia remained. Chromosome condensation and breakage in germ cells were observed under transmission electron microscope in denervated testes. TUNEL showed that SSN denervation induced apoptosis of spermatogonia and Leydig cells significantly. CONCLUSION: The superior spermatic nerve seems to be involved in the regulation of spermatogenesis. Further study is worth doing to clarify the possible mechanism of action. PMID- 12910691 TI - [Effect of FGF-BP on angiogenesis in squamous cell carcinoma]. AB - OBJECTIVE: To localize fibroblast growth factors binding protein (FGF-BP) mRNA in squamous cell carcinoma (SCC) and study the FGF-BP association with vascularity. METHODS: Thirty-five primary squamous cell carcinoma specimens, 8 metastatic squamous cell carcinoma specimens and 7 histologically adjacent tissues were studied. The localization of FGF-BP mRNA expression was performed using in situ hybridization with 35 S-labeled human FGF-BP. Microvessels in tumor specimens were identified by von Willebrand Factor-specific immunohistochemical staining for endothelial cells. RESULTS: In situ hybridization demonstrated that FGF-BP mRNA was overexpressed in SCC. But a specific hybridization signal for FGF-BP mRNA was not detected in normal adult muscle, larynx, tongue, and lymph node. The highest density of microvessels was observed at the invasive edge of the tumor. The mean microvessel count in FGF-BP positive SCC was significantly higher than that in FGF-BP negative tumors (P < 0.001). CONCLUSION: The results demonstrated that FGF-BP is present in SCC of the head and neck. The association of FGF-BP expression with angiogenesis suggests that FGF-BP may be an angiogenic factor in SCC. PMID- 12910692 TI - [Inhibition effect of aspirin on the growth of gastric cancer and the mechanism there-in involved]. AB - OBJECTIVE: To investigate the effect of aspirin on the growth of gastric cancer in vitro and the relevant mechanism. METHODS: The effect of aspirin on proliferation of SGC-7901 cells was measured by 3H-thymidine incorporation into DNA; the cell cycle was analyzed by flow cytometry. COX-2 protein was examined in SGC-7901 cells by immunohistochemistry. The expression of c-fos and the AP-1 activity were detected by immunoblotting and EMSA respectively. RESULTS: Aspirin decreased 3H-thymidine incorporation into SGC-7901 cells. The inhibition effect showed a good correlation with aspirin over a range of concentrations from 1 x 10(-1) mol/L to 1 x 10(-5) mol/L (r = 0.9, P < 0.01). Immunohistochemistry and immunoblotting indicated that aspirin effectively decreased COX-2 and c-fos expression in SGC-7901 cell. Aspirin could inhibit the AP-1 binding activation stimulated by fetal calf serum. CONCLUSION: Aspirin is effective for inhibiting the growth of gastric cancer. The anti-neoplastic effect aspirin produces may involve the inhibition of COX-2 expression and AP-1 activity. PMID- 12910694 TI - [Detection of point mutation in p53 gene by capillary electrophoresis-PCR-SSCP analysis]. AB - OBJECTIVE: To screen the point mutation of p53 gene rapidly by capillary electrophoresis (CE). METHODS: A simple capillary electrophoresis system for the strand conformational polymorphism (SSCP) analysis of genomic DNA was developed by choosing commercially available capillary and gel buffer. This CE-SSCP system was applied to the analysis of twenty PCR products amplified from the exon 7 of the p53 gene of tissue specimens from patients with colon cancer. RESULTS: Five of them (Ca4, Ca6, Ca7, Ca8, Ca14) were found to have mutation within 25 minutes, while the results of the SSCP analysis by PAGE-silver staining techniques only detected mutation of four samples. And sequencing of these PCR products proved that the results of CE-SSCP were correct. CONCLUSION: This simple, accurate and less expensive method could be used in the field of medical research and in the clinical DNA diagnosis of human cancers and other diseases. PMID- 12910693 TI - [A preliminary study of screening assay for thyroid hormone disruptors in vivo]. AB - OBJECTIVE: To study the biological effects of thyroid induced by N',N-methylene bis (2-amino-5-sulfhydryl-1, 3, 4-thiadianole), discussing whether this agent is a thyroid hormone disruptor. METHODS: Sixty Wistar rats were divided into 2 groups, including one experiment group and one control. The rats of experiment group were fed with this agent at 50 mg/kg. Whole animals were weighed, then they were sacrificed by bleeding from femoral artery on the fifth, 10th, 20th day, respectively; at the same time, serum T4 and TSH concentrations were examined and the thyroids were taken out and weighed for coefficient of thyroid gland weight; the pathological changes of thyroids were observed under light microscope; the enzyme activity of succinate dehydrogenase and thyroidperoxidase, and the positive expression of PCNA in thyroid gland were examined by histochemistry. RESULTS: Increased coefficient of thyroid gland weight, proliferation of thyroid epithelium, increased enzyme activity of SDH and TPO, and increased positive cells of PCNA were observed in the experiment group. CONCLUSION: N',N-methylene bis (2-amino-5-sulfhydryl-1, 3, 4-thiadianole) is likely to be a thyroid hormone disruptor. PMID- 12910695 TI - [The effects of selective cyclooxygenase-2 inhibitors on the growth of gastric adenocarcinoma]. AB - OBJECTIVE: This study was aimed to compare the effects of three kinds of selective cyclooxygenase-2 inhibitors (meloxicam, celecoxib, rofecoxib on the growth of gastric adenocarcinoma SGC7901 cell line, and to observe the effect of rofecoxib, on transplanted gastric cancer of nude mice in vivo. METHODS: The proliferation and apoptosis of SGC7901 cells were measured by 3H-thymidine incorporation into DNA and the TdT-mediated dUTP nick end-labeling assay (TUNEL) separately. The expression of PCNA and COX-2 of gastric adenocarcinoma cells were detected by immunocytochemistry. Human gastric adenocarcinoma SGC7901 cells were implanted orthotopically in the stomach of nude mice. Rofecoxib (30 mg.kg-1.d-1) was administrated i.g. for eight weeks. RESULTS: All the drugs potentially decreased 3H-thymidine incorporation into SGC7901 cells. The inhibition effects showed a dose-dependence manner. The median-response concentration was: 1.18 x 10(-7) mol/L (meloxicam), 1.68 x 10(-8) mol/L (celecoxib), 4.39 x 10(-9) mol/L (rofecoxib). After treatment with meloxicam, celecoxib, rofecoxib (1 x 10(-5) mol/L) for 24 hours, the apoptosis indices of SGC7901 cells were: 19.8% +/- 1.8%, 24.6% +/- 1.2% and 31.2% +/- 2.2%, respectively. The higher selective inhibition on COX-2, the higher apoptosis index (P < 0.01). Rofecoxib down-regulated the expression of COX-2 and PCNA of SGC7901 cell, both in vitro and in vivo. The inhibition rate for xenografts in situ in nude mice treated with rofecoxib was 93.9%. CONCLUSION: The higher selective inhibition on COX-2, the stronger inhibition on gastric adenocarcinoma cells. Rofecoxib may be one of the important medicines in the treatment of gastric adenocarcinoma. PMID- 12910696 TI - [Relationship of the expression of CD44v6 and paxillin to the prognosis of non small cell lung carcinoma]. AB - OBJECTIVE: To investigate the expression and significance of CD44v6 and Paxillin in non-small cell lung carcinoma. METHODS: Immunohistochemical staining was used to detect the expression of CD44v6 and Paxillin in 79 cases of non-small cell lung carcinoma, and the relationships of the expression with histological type, lymph node metastasis and prognosis were detected. RESULTS: The expression rates of CD44v6 and Paxillin were 60.8% and 34.1% respectively. The expression of CD44v6 was positively correlated with metastasis and negatively correlated with the 2-year survival rate (P < 0.05). The expression of Paxillin was positively correlated with metastasis (P < 0.05). CONCLUSION: The expression of CD44v6 and Paxillin is closely correlated with the lymph node metastasis and the prognosis of non-small cell lung carcinoma. PMID- 12910697 TI - [Studies of hTERT gene expression in acute leukemia children]. AB - OBJECTIVE: To explore the gene expression of human telomerase reverse transcriptase (hTERT) in acute leukemia (AL) children and its clinical implication. METHODS: Expression of hTERT mRNA was detected in K562 leukemia cell line, 40 acute leukemia children (AL) and 10 healthy subjects by reverse transcriptase-polymerase chain reaction (RT-PCR). RESULTS: hTERT gene was expressed in K562 cell line, acute lymphoblastic leukemia children (23/30), acute myeloid leukemia children (7/10), and healthy subject (1/10). The expression level of hTERT gene was observed in all subtypes of AL and was related to the prognosis of AL, and the patients with relative level > or = 1.0 were found to have lower complete remission (CR) rates and disease-free survival. CONCLUSION: hTERT gene expression is associated with the pathogenesis of childhood AL. It can serve as a prognostic indicator in childhood AL. PMID- 12910698 TI - [Study on the chromosome aberration of peripheral blood lymphocytes in patients with nasopharyngeal carcinoma]. AB - OBJECTIVE: To study the peripheral lymphocyte chromosome aberration in the patients with nasopharyngeal carcinoma. METHODS: The peripheral blood lymphocytes, from 28 patients with nasopharyngeal carcinoma and 20 normal controls were cultured with TC199 medium for this study on chromosome aberration. RESULTS: The chromosome number aberration rate, structure aberration rate, micronucleus rate and cell aberration rate of the peripheral lymphocytes from the patients with nasopharyngeal carcinoma were 8.68%, 10.89%, 1.83% and 8.65%, and these rates were significantly higher than those of normal controls, respectively (P < 0.05 or P < 0.01). CONCLUSION: The above findings demonstrated the instability of chromosomes in the peripheral blood lymphocytes of nasopharyngeal carcinoma patients. PMID- 12910699 TI - [Effect of para-aminobenzonic acid on cell-surface hydrophobicity of Actinomyces viscosus]. AB - OBJECTIVE: To study the effect of Para-aminobenzonic acid (PABA) on cell-surface hydrophobicity of Actinomyces viscosus. METHODS: Microbial adhesion to hydrocarbons (MATH) was used to measure the cell-surface hydrophobicity of Actinomyces viscosus which grew in modified Carlsson medium with different dilution of PABA. RESULTS: Following the increase of concentration of PABA, the value of cell-surface hydrophobicity of Actinomyces viscosus decreased, which were 0.38900 +/- 0.00026, 0.25462 +/- 0.00168, 0.16360 +/- 0.00026 respectively(P < 0.05). CONCLUSION: PABA could inhibit the adherence of Actinomyces viscosus by way of changing its cell-surface hydrophobicity. PMID- 12910700 TI - [The influence of gene vaccines pcDNA3-pac and pcDNA3-gtfB of Streptococcus mutans on plaque index and S. mutans score in rats]. AB - OBJECTIVE: This study was aimed at using the gene vaccines pcDNA3-pac and pcDNA3 gtfB which were constructed on PAc and GTF-B gene vaccinating gnotobiotic rats by submandibular gland-target injection to observe the level of salivary S. mutans and dental plaque inhabitation. METHODS: Thirty-six 28-day-old Wistar rats were divided into 6 groups, including those vaccinated with pcDNA3-pac, pcDNA3-gtfB, pcDNA3-pac combined with pcDNA3-gtfB, inactive whole cell of S. mutans JBP, and those injected with pcDNA3 and 0.1 mol/L PBS buffer respectively. Each rat was administrated with 100 micrograms vaccines or controlled materials by submandibular gland-target injection, vaccinated three times at 1-2 weeks interval. Then the gnotobiotic rat model was constructed, following that all rats were fed with cariogenic diet 2000# for 3 months. Rat saliva was collected to assay the level of S. mutans in saliva and evaluate the dental plaque index. RESULTS: The level of S. mutans in saliva and the dental plaque index of the group vaccinated with pcDNA3-pac combined with pcDNA3-gtfB and the group with inactive S. mutans cell were lower than those of other groups. And they were the highest in the groups injected with pcDNA3 or PBS buffer (P < 0.01 or P < 0.05). No statistically significant differences were observed between the group vaccinated with pcDNA3-pac and the group vaccinated with pcDNA3-gtfB, between the group vaccinated with pcDNA3-pac combined with pcDNA3-gtfB and the group for positive control, and between the pcDNA3 group and PBS buffer group (P > 0.05). CONCLUSION: The two gene anticaries vaccines induce the specific antibodies and thus produce effects on the level of S. mutans in saliva and dental plaque index. Therefore gene vaccines pcDNA3-pac and pcDNA3-gtfB are effective, and joint gene vaccines inoculation is optimal. PMID- 12910701 TI - [Animal experiment on the histological changes and associated factors of autotransplanted costochondral graft in arthroplasty of TMJ]. AB - OBJECTIVE: To explore the histological changes and associated factors of autotransplanted costochondral graft in arthroplasty of the temporomandibular joint(TMJ). METHODS: Twenty two New Zealand rabbits of 3 months were randomly divided into 4 groups. Each of A, B, C groups had 6 animals. The left discs, condyloid processes and upper parts of the mandibular rami were removed; 1 cm of rami in A, B groups and 2 cm of rami in C group were resected. The lengths of the grafts were all 2 cm. Arthroplasty with autotransplanted costochondral graft was modeled in A, B groups but the lengths of the chondral parts were different, with 4 mm in group A and 1 mm in group B. In group C the lengths of chondral parts were also 4 mm but the grafts were fixed in deep soft tissue where the rami had been taken off. Group D comprised 4 animals as normal control. The rabbits were sacrificed in batches at weeks 4 and 6 after operation. Cephalometrical and histological examinations were conducted. RESULTS: X-ray films showed the heights of the left mandibles were shorter than those of the right side in groups A, B and C, but significant difference was observed only between groups B and C. Histological examination showed that bony connections had been formed between the bony part of the grafts and the rami in groups A and B, with proliferative layer of the cartilage approaching to articular surface and on which parallel dense collagenous fibers could be found. Group B had less cartilaginous proliferation and regeneration than Group A. The chondral part of group C displayed hypertrophy, surounded by newly formed cartilage. But the growth centers in the costochondral junctions of the grafts still kept intrinsic structural characteristics. CONCLUSION: Remodeling of the costochondral grafts appeared in groups A, B and C. Nevertheless adaptable remodeling only developed in the group with the grafts firmly fixed to the mandibular rami. The shorter the length of chondral parts, the less the ability of cartilaginous proliferation and regeneration. PMID- 12910702 TI - [Effect of chondroitin sulfate-A attached titanium surface on culture osteoblast]. AB - OBJECTIVE: To investigation the in vitro effect of CS-A attached titanium surface on the culture of osteoblast. METHODS: We examined the cell proliferation, the ALP activities, and the Calcium contents in the Ostelblast cultured on the Titanium surfaces attached with different concentrations of CS-A Calcium. RESULTS: The CS-A promots the proliferation of osteoblast and increases the calcium content on the titanium disks. CONCLUSION: The biocompatibility of titanium was improved after CS-A attached titanium surface. PMID- 12910703 TI - [Effect of Radix scutellariae on the growth and form of Porphyromanus endodontics in vitro]. AB - OBJECTIVE: To provide scientific evidence for underpinning the use of Radix Scutellariae in the treatment of pulp and periapical diseases. METHODS: Using minute amount serial dilution test, this study assessed the minimal inhibitory concentration of Radix Scutellariae extreact against Porphyromanus Endodontics. The forms of Porphyromanus endodontics treated with Radix Scutellariae extract at different concentrations were observed by use of SEM. RESULTS: The MIC of Radix Scutellariae extract was determined to be 1 mg/ml. Radix Scutellariae changed the forms of Porphyromanus endodontics, making them become bigger, longer and crumbled. CONCLUSION: Radix Scutellariae could inhibit the growth of Porphyromanus Endodontics. PMID- 12910704 TI - [The effect of Galla chinensis on the demineralization of enamel]. AB - OBJECTIVE: To investigate the effect of Galla Chinensis with or without fluoride on demineralization of enamel. METHODS: Galla Chinensis with or without fluoride and acid buffer were used to treat the samples alternately and the treatment was repeated 8 times. Then the calcium of acid buffer was measured by American PE HTS 7000 plus and the specimens were observed with laser scanning confocal microscope. RESULTS: Compared with the calcium concentration of control group (deionized water or fluoride), the calcium concentration of all tested groups (except the 1 mg/ml group without fluoride) decreased significantly (P < 0.25). There was marked difference in total fluorescence (TF) and average fluorescence (AF) between the tested groups and control group (P < 0.05). As for the area of lesion, only groups without fluoride and the 4 mg/ml group containing fluoride decreased (P < 0.01). Confocal images showed that the zone of red area of tested groups was narrower than that of control group. CONCLUSION: Galla Chinensis can reinforce the resistance of enamel to acid, either with or without fluoride. PMID- 12910705 TI - [The effect of rehabilitation of flared root canal on the fracture resistance of root and post-core system]. AB - OBJECTIVE: To assess the effect of rehabilitation of flared root canal on the fracture resistance of root and post-core system. METHODS: Thirty-two simulated tooth roots made of polymethyl methacrylate were divided into 4 groups, including the normal root canal group and 3 flared root canal groups. One flared root canal group was rehabilitated by light-cared composite, another group was rehabilitated by amalgam, and the third flared root conal group was not treated. Casting post and core restored every simulated tooth root. Each specimen was embedded in acrylic resin and then fixed in a special jig on the universal load-testing machine. A compressive load was applied at a 90-degree angle to the long axis of the core until fracture, at a crosshead speed of 1 mm/min; the maximum of load was recorded. RESULTS: The means of load in normal root canal group, light-cared composite rehabilitated group, amalgam rehabilitated group, not treated group were 202.92 N, 194.60 N, 146.89 N, 142.09 N. It was shown that the load of normal root canal group is significantly greater than the load of not-treated group (P < 0.01); the light-cared composite rehabilitated group is significantly stronger than the not-treated group (P < 0.01), but there is no significant difference between the light-cared composite rehabilitated group and normal root canal group, and no significant difference between the amalgam rehabilitated group and not-treated group (P > 0.05). CONCLUSION: The rehabilitation of flared root canal using suitable material not only changes the shape of root canal, but also increases the fracture resistance of root canal because of its thicker and stronger root canal wall which obviously improves the fracture resistance of post and core. PMID- 12910706 TI - [The age-related changes of P53 protein expression in oral mucosa and their effects on oral leukoplakia]. AB - OBJECTIVE: To investigate the age-related changes of P53 protein expression in oral mucous membrane and their effects on pathogenesis of the oral precancerous lesions in elderly people. METHODS: An immunohistochemical technique (SP method) was applied to study the changes of P53 expression in normal oral mucosa and oral leukoplakia(OLK) in different age groups. The staining results were evaluated by semi-quantity method. RESULTS: With aging, an elevating tendency of P53 protein level exhibited in the normal oral mucosa and OLK, respectively. The P53 level in the old with OLK was higher than that in the normal oral mucosa of the old (P < 0.01). CONCLUSION: With aging, P53 protein may undergo dysfunction, lead to imperfect DNA repair and excessive DNA damage accumulation, and ultimately facilitate malignant transformation of the oral mucosa in the old. PMID- 12910707 TI - [Effects of asbestos on P53 protein expression in V79 cells]. AB - OBJECTIVE: To explore the effects of physical and chemical properties of asbestos on mutagenesis/carcinogenesis. METHODS: The P53 protein expression in V79 cells exposed to asbestos suspension and filtrate was studied using immunohistochemical method. RESULTS: After asbestos suspension and filtrate treatment of V79 cells, the value of hue for indicating P53 protein expression was significantly lowered, and such abnormal expression was especially observed in binucleate cells and polykaryocytes. CONCLUSION: The above data demonstrated that abnormal accumulation of P53 protein in V79 cells was related to asbestos exposure and that asbestos suspension and filtrate had even greater effect on the cells at the segmentation period. PMID- 12910708 TI - [Estimate of human visceral adipose area and abdominal subcutaneous adipose area in obese Chinese by simple anthropometric parameters]. AB - OBJECTIVE: To explore the relationship of simple anthropometric parameters with abdominal subcutaneous adipose area (SA) with visceral adipose area (VA), and to generate equations predicting SA and VA by simple anthropometric parameters. METHODS: SA and VA were measured with computed tomography (CT) in 309 human subjects (male 88, female 221). Weight (W), height (H), body mass index (BMI), waist circumference (WC), abdominal circumference (AC), hip circumference (HC) and waist to hip ratio (WHR) were also measured. Multiple stepwise regression analysis was used to generate equations for predicting SA and VA from age and simple anthropometric parameters of 259 subjects (80%, including 181 women and 78 men) randomly selected from the overall sample. These equations were then cross validated in the remaining 50 subjects (20%, including 40 women and 10 men). RESULTS: The best regression equations for male were developed for predicting SA and VA, and the explanatory variables included WC and age. In women, the equation for predicting SA included AC and BMI; the equation for predicting VA included WHR and W and age. In the cross-validation study, the differences between predicted and observed values of VA in men and women were -7.83% and -6.94%, respectively; the differences between predicted and observed values of SA in men and women were 8.01% and 0.69%, respectively. The goodness of fit between predicted and observed values is good. CONCLUSION: The absolute amount of human VA and SA in obese Chinese can be predicted from anthropometric measurements. PMID- 12910710 TI - [Establishment of whole pancreaticoduodenal allotransplantation model with jejunum drainage and portal venous drainage in pigs]. AB - OBJECTIVE: To establish a pig model of pancreaticoduodenal transplantation with jejunum drainage and portal venous drainage. METHODS: Forty pigs were randomly divided into the donor group and recipient group. University of Wisconsin (UW) solution was used for both in situ flush and storage of all organs under cold storage conditions. Whole pancreaticoduodenal graft was harvested with segments of abdominal aorta and portal vein and shaped under cold UW solution. Then, the end-to-end anastomosis was performed with the donor iliac artery bifurcation Y graft to the recipient superior mesenteric arteries and celiac artery. Furthermore, type I diabetes model was made by removal of the recipient pancreas The venous anastomosis was reconstructed between the donor portal vein and the recipient superior mesenteric vein. Meanwhile, the end-to-side anastomosis was performed with the donor common iliac artery bifurcation Y graft to the recipient abdomial aorta, and the side-to-side intestinal anastomosis between the donor duodenum and the recipient jejunum was performed. RESULTS: Twenty piges were subjected to pancreaticoduodenal transplantation, in which 1 died from respiratory depression because of anesthesia, and 2 had thrombus and infarction in the pancreatic graft after transplantation. 17 cases showed normal blood sugar in peripheral circulation 24 hours after operation, the mean survival of the graft was 18.6 +/- 2.6 days. CONCLUSION: The technique of duodenal transplantation with jejunum drainage and portal venous drainage may pave the way for the further development of pancreas transplantation in clinical practice. It can be utilized in basic research of pancreas transplantation. PMID- 12910709 TI - [Effect of Fos expression induced by spinal nitric oxide on the nociception of formalin in rats]. AB - OBJECTIVE: To assess the effect of s.c. injected formalin and clarify the possible mechanism of nociception in SD rats. METHODS: Sixty-four healthy male rats were randomly distributed into the control groups (group NS, group formalin) and test groups (group LaF, group LnF) with correponding numbers. Group NS served as normal control. Group formalin was treated with 5% formalin 100 microliters, s.c. Groups LaF and LnF were treated respectively with L-Arg 4.7 mumol ith and L NAME 370 nmol ith before receiving 5% formalin 100 microliters, s.c. like group formalin. 1/2 h and 0-1 h after formalin treatment, spinal cord segments were removed for expression of NOS, and behavior responses were observed. RESULTS: The flinching and licking time of group formalin was longer than that of group NS, and NOS expression of group formalin was stronger than that of group NS. Pretreatment with L-Arg potentiated formalin-induced response, but pretreatment with L-NAME reduced the response. CONCLUSIONS: Spinal nitric oxide release might be one of the nociception mechanisms in the formalin test. PMID- 12910711 TI - [Human leucocyte antigen-DPA1 polymorphism distribution in Chengdu Chinese families with intrahepatic cholestasis of pregnancy]. AB - OBJECTIVE: To investigate the relationship between human leucocyte antigen (HLA) DPA1 region gene polymorphism and intrahepatic cholestasis of pregnancy (ICP). METHODS: The HLA-DPA1 alleles of 25 ICP families and 25 normal control group families were analysed by polymerase chain reaction-sequence specific primers(PCR SSP) method. RESULTS: The analysis of the frequencies and feto-maternal or couples sharing of the DPA1 alleles showed that no statistically significant difference was observed between the two study groups. CONCLUSION: It is unlikely that an HLA-DPA1 allele is primarily associated with susceptibility or resistance to ICP. PMID- 12910712 TI - [An epidemiologic investigation of acute rheumatic fever and rheumatic heart disease among students aged 5-18 in west area of Sichuan Province]. AB - OBJECTIVE: To study the incidence of acute rheumatic fever (ARF) and its epidemiologic features and the prevalence rate of rheumatic heart disease in west area of Sichuan province. METHODS: By using disease surveillance program, an epidemiologic investigation of ARF was performed from June 1992 to June 1995 and an annual screening survey was carried out among 46,595 students aged 5-18 in 1993 and 1994. RESULTS: The annual incidence of ARF was 12.87 per 10(5), there was no difference between the rural area and the urban area. ARF was found to be more frequent in students aged 9-15 years and in autumn and winter. All of the ARF patients had rheumatis and 83.3% of the cases were accompanied with heart lesion. The prevalence rate of rheumatic heart disease was 11 per thousand. CONCLUSION: The long-term monitoring of epidemiologic trend and community-based controls of rheumatic fever and rheumatic heart disease among students aged 5-18 are necessary and effective. PMID- 12910713 TI - [The status quo of perimenopausal and postmenopausal women in Chengdu: report of a population-based cross-sectional study]. AB - OBJECTIVE: To investigate the situation of reproductive health of perimenopausal and postmenopausal women in Chengdu and provide a basis for planning reproductive health care for these women. METHODS: We conducted a population-based cross sectional study in women who were over 45 years old in the central city and the nearby county of Chengdu. RESULTS: The mean age of menopause was 48.1 years. There were sixty percent of women who had dysfunctional uterine bleeding before menopause. About half of the postmenopausal women had experienced perimenopausal symptoms. There were 460 women who ever used or currently use estrogen therapy. But only half of them visited doctors and sought medical therapy. And the main problems were hot flash, night sweats, dyssomnia, and urinary infection. Most of them took medicine for a short period (< 1 year). CONCLUSION: We found that middle-aged and older women have many symptoms related to menopause. It is necessary for us to provide reproductive health care for them. PMID- 12910714 TI - [Stereopsis of the patients after cataract extraction and intraocular lens implantation]. AB - OBJECTIVE: To understand the stereopsis of the patients after cataract extractation and intraocular lens implantation and the related factors. METHODS: In this study were examined the stereopsis of 30 patients who had undergone cataract extraction and intraocular lens implantation and whose post-operative vision acuity measurements were 0.5 or higher. RESULTS: These patients' steroacuity, anisometropia, aniseikonia and scotoma were: 407.67" +/- 390.19", 1.53D +/- 1.96D, 3.93% +/- 3.51%, 3.17" +/- 1.98" respectively. There was no significant correlation between steroacuity anisometropia and aniseikonia. There was siganificant positive correlation between steroacuity and scotoma. CONCLUSIONS: Intraocular lens implantation may ameliorate anisometropia and aniseikonia of the patients after cataract extraction and reduce the damage to stereopsis. PMID- 12910715 TI - [Pharmacokinetics and relative bioavailability of KC-404 sustained release tablets in healthy volunteers]. AB - OBJECTIVE: To study the pharmacokinetics and relative bioavailability of KC-404 sustained release tablets and capsules in healthy volunteers. METHODS: The concentration of KC-404 in serum was determined by HPLC method after a single oral dose (20 mg) of tablet or capsule was administered to 19 healthy male volunteers respectively in an open randomized cross-over test. RESULTS: After being processed by 3P87 pharmacokinetics program, the experiment data showed that the pharmacokinetic parameters of the tablets and the capsules were AUC0- >infinity: 740.12 ng.h/ml and 724.04 ng.h/ml; tmax: 5.37 h and 5.11 h; Cmax: 46.98 ng/ml and 46.29 ng/ml; T1/2: 7.4 h and 7.0 h; MRT0-->infinity: 14.78 h and 14.39 h respectively. There were no significant differences in AUC, tmax, Cmax, T1/2 and MRT0-->infinity between these two preparations (P > 0.05). The relative bioavailability of KC-404 sustained release tablets was 102.6%. CONCLUSION: These two preparations are bioequivalent. PMID- 12910717 TI - [Effect of ginsenoside Rg3 on the progression of orthotopically xenotransplanted human breast cancer in nude mice and its mechanism]. AB - OBJECTIVE: To explore the inhibiting effect of ginsenoside Rg3 on the growth and angiogenesis of orthotopically xenotransplanted human breast infiltrating duct carcinoma in nude mice. METHODS: A total of 15 female nude mice having received xenotransplanted human breast infiltrating duct carcinoma were randomly divided into 3 groups. Ginsenoside Rg3 (5 mg/kg, qd), cyclophosphamid (26 mg/kg, qod) and control solution (0.5% sodium carboxymethyl cellulose, qd) were given to each group by gas-trogavage in 0.5 ml volume for 56 days. Breast cancer masses were collected for light microscope observation. The intra-tumoral microvessel density (MVD) and the expression of vascular endothelial growth factor(VEGF) were examined by immunohis-tochemical staining. RESULTS: The tumor weight of CTX group was significantly lower than that of control group (P < 0.05), and the comparison between those of Rg3 group and control group was nearly reaching significant level. The MVD and VEGF of Rg3 group by immunohistochemical staining were significantly lower than those of control group (P < 0.05). CONCLUSION: Rg3 could inhibit the growth and angiogenesis of xenotransplanted human breast infiltrating duct carcinoma in nude mice. PMID- 12910716 TI - [Effects of NaBT and NSAIDs on colorectal adenoma-carcinoma cells]. AB - OBJECTIVE: We investigated the primary adenoma cells and HT-29 cells in an attempt to know whether NaBT and two NSAIDs (aspirin and NS-398) could be used as effective adjuvant for reducing cell proliferation, and to explore the possible mechanisms therein involved. METHODS: MTT was employed in detecting the anti proliferative effects on both adenoma and HT-29 culture cells. RESULTS: Both butyrate and NSAIDs could inhibit the proliferation of cells. The effects of the NSAIDs could be synergistically strengthened by combining with butyrate. CONCLUSION: NaBT and the two NSAIDs could inhibit the cell proliferation when used separately, and a combination of butyrate with aspirin or NS-398 could enhance the inhibitory effects. PMID- 12910719 TI - [Short-term effects of cyclosporine A on the function of pancreatic islet cells in normal and spontaneous diabetic Chinese hamsters]. AB - OBJECTIVE: To examine the effects of low dose (25-100 ng/ml) CsA on the hormonal secretion of the pancreatic islet cells isolated from normal and spontaneously inherited diabetic Chinese hamsters. METHODS: A modified kanatsuna and Knudsen's column perifusion system was developed. The secretions of Insulin (Ins) and glucagon (Glc) of the cultured islet cells responding to the glucose stimulations were tested after pretreated with different doses of CsA for 24 hours. RESULTS: The secretions of Ins of the diabetic islet cells with and without glucose stimulations (16.7 mmol/L) both reduced significantly compared to the normal controls (P < 0.05). In contrast, the static secretions of Glc increased significantly in the diabetes islet cells (P < 0.05). The response (Ins release) of normal islets to glucose stimuli was significantly inhibited after 24-hour pretreatment with CsA (25-100 ng/ml, P < 0.05), while the concentrations of Ins and Glc detected in the culture fluids did not differ from the controls (P > 0.05). CONCLUSION: Abnormal islet function exists in diabetic hamsters characterized by impairment of Ins secretion and increase of Glc secretion. Low dose CsA has no significant effects on the secretions of Ins and Glc, but it inhibits the response of the islets to the glucose stimuli. PMID- 12910718 TI - [The variation of Caspase3 activity in tanshinone induced NB4 cells apoptosis]. AB - OBJECTIVE: To study the variation and role of Caspase3 activity in the process of Tanshinone (Tan II A) induced NB4 cells apoptosis. METHODS: NB4 cell apoptosis induced by Tan II A was demonstrated by cell morphology, DNA content analysis and DNA fragmentation assay. Caspase3 activity was determined by spectrofluorometry, and its inhibitory assay was performed using N-acetyl-Asp-Glu-Val-Asp-aldehyde(AC DEVD-CHO). RESULTS: Tan II A could induce NB4 cell apoptosis accompanied with increase of caspase3 activity. The induction of NB4 cell apoptosis by use of Tan II A could be partially inhibited by AC-DEVD-CHO. CONCLUSION: The induction of NB4 cell apoptosis by Tan II A could be fulfilled by activating Caspase3. PMID- 12910720 TI - [Preventive effects of montelukast on hypoxic pulmonary hypertension in rats]. AB - OBJECTIVE: To evaluate the role of cysteinyl leukotriene(CysLT) in the pathogenesis of hypoxic pulmonary hypertension and the preventive effects of montelukast, a CysLT receptor antagonist, on hypoxic pulmonary hypertension. METHODS: Thirty male Wistar rats were randomly divided into three groups: control group, hypoxic group and montelukast preventive group. The animal model of pulmonary hypertension was established by exposing the rats to normabaric hypoxic conditions for 3 weeks. The thickness of pulmonary arterioles was measured by a computerized image analyzer. The level of LTC4 in plasma was measured by EIA. RESULTS: In the hypoxic group, the index of right ventricular hypertrophy[RV/(LV + S)] and the index of wall thickness of pulmonary arteriole (WT% and WA%) increased significantly when compared against those in the control group [RV/(LV + S): 30.85% +/- 1.44% vs 20.75% +/- 1.97%; WT%: 23.54% +/- 4.43% vs 13.17% +/- 3.67%; WA%: 72.76% +/- 9.28% vs 50.41% +/- 6.37%, P < 0.01, respectively]. Meanwhile, the plasma level of LTC4 in hypoxic rats was higher than that in control rats [(2395.40 +/- 193.86) pg/ml vs (1006.50 +/- 193.17) pg/ml, P < 0.01]. In the preventive group, the RV/(LV + S) (24.09% +/- 1.09%), WT% (15.44% +/- 4.72%) and WA% (51.98% +/- 12.18%) decreased remarkably as compared with those of the hypoxic group(P < 0.01). The plasma level of LTC4 in the preventive group (2706.25 +/- 350.49 pg/ml) was not significantly different from that in the hypoxic group(P > 0.05). CONCLUSION: Chronic hypoxia stimulates synthesis and release of LTC4. CysLT may play an important role in the pathogenesis of hypoxic pulmonary hypertension and montelukast can prevent hypoxi pulmonary hypertension. PMID- 12910721 TI - [Research on motor dysfunction and the role of CTP after traumatic brain injury in rats]. AB - OBJECTIVE: This is a study on the level of molecular biology to clarify the possible mechanism of delayed neural death after traumatic brain injury (TBI) and to probe into the influences of motor function of cytidine triphosphate(CTP) on rats after brain trauma. METHOD: The model of severe closed traumatic brain injury (TBI) was established according to the method created by Marmarou in 1994. 300 Wistar rats were divided randomly into TBI group (n = 96), CTP treating group and sham operation group, and each of these three groups was divided into 8 subgroups, namely, 3, 6, 12, 24, 48, 72, 168 and 336 hours. At the same time, the rest 12 rats were taken as the normal group for comparison. The CTP treating group after injury were treated with injection of CTP (30 mg/(kg.d)) in peritoneum; the TBI group, the sham operation group and the normal group were treated with normal saline for comparison at the same time and all were killed at each time point. Another 48 rats were divided randomly into 4 groups, namely, brain injury group, CTP treating group, sham operation group and the normal group, and their motor dysfuction was evaluated using Neurological Severity Score (NSS). The methods such as immunohistochemistry, TUNEL and NSS were used to dynamically observe the pathological changes in cortex of rats, the expression of P53 protein and the motor function after TBI. RESULTS: The increased expression of P53 protein and neural apoptosis showed up in cortex and motor dysfunction occurred after TBI. CONCLUSION: The increased expression of P53 protein may bring about nerve cell apoptosis after TBI. Motor dysfunction is the inevitable outcome of nerve cell apoptosis in cortex. CTP can reduce P53 protein expression and nerve cell apoptosis, and it can improve motor function. PMID- 12910722 TI - [Investigation of the effect of Na-butyrate on TNBS induced rat colitis]. AB - OBJECTIVE: To observe the effect of Na-butyrate on trinitrobenzene sulfonic acid (TNBS) induced rat colitis and discuss its mechanism. METHODS: 0.85 ml of 50% ethanol enema containing 30 mg TNBS was instillated into rat colon to produce distal colitis. On the 5th day after TNBS enema, the rats were divided into 3 groups and were given respectively 100 mmol/L Na-butyrate, 100 mg/kg SASP or 0.9% saline enema daily. On the 19th day, macroscopical, histological injury scores of colonic mucosa and tissue myeloperoxidase (MPO) activity were evaluated, and the mitotic index in Na-butyrate group was compared with that in SASP group. RESULTS: Diarrhea was apparently improved, weight gain was not apparently different, the scores of macroscopical and histological injury and MPO activity decreased in the SASP group and the Na-butyrate group, compared with the control (P < 0.05). However, there was not obvious difference between the Na-butyrate group and SASP group (P > 0.05). Mitotic index in Na-butyrate group was higher than that in SASP group (P < 0.05). CONCLUSION: Na-butyrate enema can expedite the restoration of the experimental colitis, and its therapeutic effect is as good as that of SASP. PMID- 12910723 TI - [The Predictive Role of Vascular Endothelial Growth Factor and Estradiol in Infertile Patients with Ovarian Hyperstimulation Syndrome]. AB - OBJECTIVE: To determine the predictive values of vascular endothelial growth factor (VEGF) and estradiol (E2) in patients with ovarian hyperstimulation syndrome (OHSS) and to go further into the relevant pathogenisis. METHODS: Blood samples were collected from 51 high risk patients and 30 control patients undergoing superovulation on the day of hCG administration and follicular fluid at the time of egg retrieval. The high risk patients were divided into two groups[OHSS group (n=11) and non-OHSS group (n=40)] VEGF and E2 were detected and then differences between the high risk groups and control group were analyzed. RESULTS: There wa no significant difference in VEGF between high risk groups and control group on the day of hCG injection (P>0.05). After hCG administration, serum VEGF concentrations rose significantly in OHSS group (P<0.01). The serum and follicular fluid VEGF levels were obviously higher than those of the control group. The follicular fluid concentrations of VEGF were significantly higher than that of serum in every group (P<0.001). E2 concentrations of OHSS group rose obviously after hCG injection (P<0.01) and were significantly higher than those of the control group (P<0.01), but there no statistically significant change in E2 concentrations in non-OHSS groups (P>0.05). CONCLUSION: The results from this study show that the serum and follicular fluid VEGF concentrations are significantly higher than those of control group on the day of ovum retrieval, indicating that VEGF may play an important role in the pathogenesis of OHSS. It cannot predict the risk of OHSS on the day hCG administration, but it may be considered as a predictive marker for the development of OHSS after hCG administration. Serum VEGF concentrations in combination with consective E2 measurements can assist in predicting OHSS. PMID- 12910724 TI - [Quantitative analysis of mixed hematopoietic chimersim based short tandem repeats loci after nonmyeloablative stem cell transplantation]. AB - OBJECTIVE: To establish a method for quantitative analysis of mixed hematopoietic chimerism (MC) based on STR loci. METHODS: Using STR technique combined with PCR, we analyzed the peripheral blood samples collected from 3 nonmyeloablative stem cell transplantation recipients every 15 days after transplantation. RESULTS: The quantitative results coincided with the qualitative results of DNA electrophoresis and with the clinical outcomes. When the result of electrophoresis showed only donor's allele type we could still find out the recipient's specific allele. CONCLUSION: The quantitative analysis using STR-PCR can detect more subtle changes in the evidence of engraftment. It is a more efficient method for detect the hematopoietic chimerism post transplantation. PMID- 12910725 TI - [On preparation the monoclonal antibody against synthetic antigen of cyclosporine A]. AB - OBJECTIVE: To study the methods for preparing the monoclonal antibody against synthetic antigen of cyclosporine A. METHODS: These included using photochemical reaction to couple cyclosporine A with poly-L-lysine as synthetic antigen, immunizing BALB/c mice, fusing spleen cells and SP2/0 cells. Then hybridoma cells were repeatedly screened and subcloned. The monoclonal antibodies were identified by indirect ELISA. RESULTS: Four hybridoma cells in secreting anti-cyclosporine A monoclonal antibodies were obtained. Only two showed high titer and some specificity. CONCLUSIONS: The methods of antigen synthesis and immunization were practicable. But the methods of antibody preparation and preservation need to be improved. PMID- 12910726 TI - [HPLC method for determination of tramadol hydrochloride in human plasma]. AB - OBJECTIVE: A reversed-phase high performance liquid chromatography (HPLC) method was established to determine the concentration of tramadol hydrochloride in human plasma. METHODS: HPLC instrument was used with the column: 150 mm x 4.6 mm (Li Chrosorb C18, 5 microns). The mobile phase was composed of phosphate buffer solution (0.02 mol/L, pH = 3.7)-acetamide (83: 17, V/V). Flow rate was 1.0 ml/min. Detection wavelength was 216 nm. RESULTS: The standard curve equation was Y = 0.03244X + 0.6007. The linear range was 25-800 ng/ml. The minimum detection limit was 10.2 ng/ml. CONCLUSION: The reversed-phase HPLC method is simple, rapid and sensitive. It is applicable to the determination of the concentration of tramadol hydrochloride in human plasma. PMID- 12910727 TI - [Determination of salbutamol in human plasma by column-switching HPLC with UV detection]. AB - OBJECTIVE: To make better the RP-HPLC method with column-switching technique for the determination of salbutamol in human plasma. METHODS: A high-pressure flow channel selection valve and Kromasil C18 pretreatment column (20 x 4 mm, 5 microns), Ultrasphere Cyano analysis column (250 mm x 4.6 mm, 5 um, Beckman) were used. To the plasma sample 1.0 ml, 0.5 ml phosphate buffer (2.0 mol/L, pH 9.0) containing 0.4% diphenylboric acid 2-aminoethyl ester was added, then extraction was performed with the use of 4.0 ml chloroform containing 1% tetraoctylammonium bromide. The organic layer was removed and extracted again with 300 microliters (0.08 mol/L) acetic acid. 100 microliters of the acid layer was injected onto the column. The mobile phase of pH 2.8, 0.025 mol/L phosphate buffer-acetonitrile methanol (95:4:1) was pumped at the rate of 0.9 and 1.0 ml.min-1 through the pretreatment and analysis column, respectively. The column-switching time was from 0.7 min to 1.5 min. The detector at 0.002 aufs was set at 224 nm. RESULTS: The retention time for salbutamol was 6.7 min and that for internal standard (morphine) 7.6 min. The standard curve was linear over the concentration range from 0.5 to 32 micrograms/L. The lowest concentration of detection in plasma was 0.5 microgram/L. The method recovery was 96%-107%; the intra-day RSD less than 5%; the inter-day RSD less than 8%. CONCLUSION: This method was found to be simple, rapid, sensitive and accurate for determination of salbutamol in human plasma. PMID- 12910728 TI - [Determination of tin in canned foods with oscillographic polarography]. AB - OBJECTIVE: To develop an oscillographic polarography method for determining the concentration of tin in canned foods. METHODS: Because the sensitive polarography wave of tin can be obtained in the system of H2SO4-KI-VitC- NH4VO3, the tin content of several canned foods was analyzed. RESULTS: The detection limit was 0.12 microgram/L. The calibration graph of Sn was linear from 0 to 2.0 micrograms/ml, and the relative standard deviation ranged from 4.22% to 8.33%. The recoveries were 94.7%-112.0%. The samples were analyzed by the proposed method and standard method. The results of the two methods agreed well. CONCLUSION: This method is simple, quick and almost free from interference, and it is available for use in analysis of the canned sample. PMID- 12910729 TI - [Isolation of cochlear inner hair cells from guinea pigs by trypsin]. AB - OBJECTIVE: To obtain isolated inner hair cells from guinea pigs by trypsin digestion and mechanical trituration. METHODS: The guinea pig was decapitated and the temporal bone was quickly removed. Then the lateral wall of the cochlea was removed. The organ of Corti was dissected out under a dissection microscope. Single hair cells were obtained by digestion with trypsin and gentle mechanical trituration. RESULTS: The isolated inner hair cells were observed under inverse microscope. Five to twenty single outer hair cells were obtained from each ear and most of them could survive for about 5 hours. CONCLUSION: This technique can provide sufficient numbers of solitary inner hair cells for studies on the electrophysiological properties of the cells. PMID- 12910730 TI - St. Luke's site upgrade improves navigation and speeds up content publishing. PMID- 12910731 TI - Customized portals may encourage users to visit your site often. PMID- 12910732 TI - Survey provides insights into how health sites can get more consumers online. PMID- 12910733 TI - Granting a reprieve. Leapfrog delays date of compliance. PMID- 12910734 TI - Connecting the supply line. Part 1. PMID- 12910735 TI - Privacy matters. PMID- 12910736 TI - Next best thing to free. Organization redistributes donated supplies. PMID- 12910737 TI - O.R. Left behind. Counting surgical instruments reduces medical errors. PMID- 12910738 TI - Update on molecular techniques for diagnostic testing of infectious disease. AB - The era of diagnostic molecular biology has arrived for small animal clinicians, and it is a near certainty that assays such as the PCR and RT-PCR will become more widely available for a wider array of infectious agents. Already there is an extensive list of infectious diseases of dogs and cats that have been investigated with molecular tools. A partial list is included in box 1. An understanding of the advantages and disadvantages of the molecular techniques and some of the questions these techniques can answer for clinicians can serve practitioners well in their approach to the diagnosis of infectious diseases in dogs and cats. It is likely that additional applications of these tools to small animal medicine will become apparent as investigators use and refine them for their research purposes, or as new uses emerge from human medical applications. Clinicians also are likely to reap the benefits of this knowledge. Because samples often are acquired easily from clinical patients in most practice settings, access to these tools puts all clinicians in the group of discoverers of new, or variations of, infectious diseases and their clinical manifestations. PMID- 12910739 TI - Pythiosis, lagenidiosis, and zygomycosis in small animals. AB - Pythiosis, lagenidiosis, and zygomycosis affect animals living in temperate, tropical, and subtropical climates, and these diseases are often fatal. Although Pythium insidiosum, Conidiobolus species, and Basidiobolus species have been recognized as pathogens for several years, members of the genus Lagenidium have been identified as a cause of oomycosis in dogs only recently. Pythiosis, lagenidiosis, and zygomycosis share similar clinical and histologic characteristics, making them difficult to distinguish from one another; however, distinguishing between these pathogens is important because of differences in epidemiology, choice and duration of therapy, and prognosis. PMID- 12910741 TI - Update on antifungal therapy. AB - Fungal pathogens are becoming increasingly important for human and small animal medicine. This article highlights many standards-of-care and new agents for treatment of these pathogens for small animals and people. PMID- 12910740 TI - Update on canine and feline fungal diseases. AB - Systemic fungal diseases cause significant morbidity and mortality in dogs and cats. Blastomycosis, histoplasmosis, coccidiomycosis, and cryptococcosis represent the four most common systemic fungal diseases. Young adult, large breed dogs generally are predisposed; cats usually do not have predictable predispositions. Intact cell-mediated immunity is essential to initial resistance to infection and response to treatment in animals. Several body systems can be affected. Diagnosis can be confirmed on the basis of clinical signs and demonstration of the causative organism. Serology is helpful with coccidiomycosis and cryptococcosis. Treatment is complicated by limited availability of fungicidal antimicrobials and the necessity of long-term treatment with expensive drugs. PMID- 12910742 TI - Update on feline calicivirus: new trends. AB - In addition to being important upper respiratory tract pathogens of cats, FCVs are increasingly reported as a cause of a highly contagious febrile hemorrhagic syndrome. Strains causing this syndrome are genetically different from the vaccine strain and other nonhemorrhagic FCV isolates. They apparently differ from one outbreak to another. The syndrome is characterized variably by fever; cutaneous edema and ulcerative dermatitis; upper respiratory tract signs; anorexia; occasionally icterus, vomiting, and diarrhea; and a mortality that approaches 50%. Adult cats tend to be more severely affected than kittens, and vaccination does not appear to have a significant protective effect. Rapid recognition of the disease through identification of clinical signs and appropriate testing, followed by strict institution of disinfection, isolation, and quarantine measures, are essential to prevent widespread mortality resulting from the infection. PMID- 12910743 TI - Feline hemotropic mycoplasmosis (feline hemobartonellosis). AB - Hemotropic mycoplasmas represent an important cause of anemia in cats worldwide. Previously known as Haemobartonella species, sequencing of the 16S rRNA genes of these organisms has led to their reclassification as mycoplasmas. Two species have been identified in cats, M haemofelis and "Candidatus M haemominutum." The latter organism alone has not been associated with disease in naturally infected cats but may cause anemia in FeLV-infected cats and accelerate development of FeLV-induced myeloproliferative disease. The mode of transmission of these organisms remains enigmatic. Nevertheless, development of sensitive DNA-based tests for these unculturable organisms has improved the understanding of the epidemiology and pathogenesis of FHM. Cats with clinical signs and laboratory abnormalities consistent with FHM should be treated with doxycycline; enrofloxacin may represent an effective alternative. Transfusion with packed red blood cells after cross-matching may be required for severely anemia cats, and addition of prednisone may be required if the diagnosis of FHM is uncertain, or response to antimicrobials alone is insufficient. Affected cats should be tested for FeLV, the most common concurrent infection in cats with FHM. PMID- 12910744 TI - Leptospirosis. A re-emerging zoonotic disease. AB - Leptospirosis is a re-emerging infectious disease that occurs in dogs in urban and rural environments. It is caused by a filamentous spiral bacterium that has a predilection for renal tubules. Acute renal failure, hepatic dysfunction, and hemorrhagic diathesis are the most common clinical signs. Treatment with antibiotics and supportive care can manage a high percentage of cases successfully. Newer vaccines developed in response to the change in frequency of certain serovars may decrease the incidence of clinical disease. Leptospirosis affects a wide variety of species and is zoonotic. PMID- 12910745 TI - Bartonellosis. AB - The role of Bartonella species as pathogens in dogs and cats is being defined. Diagnosis and treatment of Bartonella infections of dogs and cats remain challenging. As new information regarding Bartonella infections of companion animals becomes available, the understanding of the pathogenesis of these infections will improve. Most Bartonella species infecting dogs and cats are zoonotic, with B henselae the most important zoonotic species. B henselae bacteremia is common in domestic cats, and cats transmit B henselae to people. Transmission of Bartonella infections among cats and dogs is believed to occur primarily by way of arthropod vectors. Control of arthropod vectors and avoiding interactions with pets that result in scratches or bites are the most effective means to prevent transmission between animals and people. PMID- 12910746 TI - Canine borreliosis. AB - A guild of organisms carried by the same vector (Ixodes ticks) in Lyme-endemic areas may be confounding the understanding of Lyme disease in dogs. A new diagnostic method, the C6 peptide test for Lyme, and serology and PCR testing for Ehrlichia, Babesia, and Bartonella species will help to sort out seroprevalence and symptomatology caused by exposure to these agents or by coinfections. In addition, Rickettsia, Leptospira, Mycoplasma species, and more could be involved in dogs diagnosed with a "doxycycline-responsive" disease. The author does not recommend treating asymptomatic Borrelia carrier dogs, but does recommend screening them for proteinuria and for exposure to other agents. A positive Lyme titer is a marker of exposure to Ixodes ticks and the agents they carry. The risk/benefit of vaccination will be understood better as the symptomatology and immunopathogenesis of Lyme disease are defined. Meanwhile, tick control is highly recommended for all dogs in Lyme-endemic areas. PMID- 12910747 TI - Ehrlichiosis and related infections. AB - Ehrlichiosis is a term that has been used to describe infection with any of a number of related intracellular, vector-borne pathogens. A recent reclassification has resulted in the transfer of several species previously known as Ehrlichia to the genus Anaplasma or Neorickettsia. Ehrlichia and Anaplasma are transmitted largely through the bite of infected ticks, while vectors for Neorickettsia include trematodes and the intermediate hosts (i.e., fish, snails, and insects) involved in the trematode life cycle. Dogs (and cats) are susceptible to infection with several of these pathogens, and veterinarians should be aware of the similarities and differences between E canis and related infections. Pets with suggestive clinical signs and laboratory abnormalities may be started on doxycycline pending specific diagnostic testing. The veterinarian practicing in endemic areas must understand the implications and limitations of serologic and molecular testing to confirm a diagnosis. For animals in endemic areas, prevention of exposure to vectors can lessen the risk of disease for pets and might lessen the potential for animals to become carriers of disease for their human companions. PMID- 12910748 TI - Canine babesiosis. AB - Canine babesiosis is a tickborne, protozoal, hemoparasitic disease that can cause varying degrees of hemolytic anemia, splenomegaly, thrombocytopenia, and fever. Babesia organisms frequently are classified as large or small. Large Babesia infections are attributed to one of three subspecies of Babesia canis. All small Babesia infections previously were attributed to B gibsoni, but molecular analysis and DNA sequencing have revealed that there are at least three small piroplasms infecting dogs. Correctly identifying the infectious agent is important for treatment planning and prognosis. PMID- 12910749 TI - American canine hepatozoonosis. AB - Hepatozoon americanum infection is an emerging tickborne disease in the southern United States. This organism causes a very different and much more severe disease than does Hepatozoon canis, the etiologic agent of canine hepatozoonosis in the rest of the world. H americanum is transmitted through ingestion of the definitive host, Amblyomma maculatum (the Gulf Coast tick). Clinical signs of American canine hepatozoonosis tend to wax and wane over time and may include lameness, weakness, pain, muscle atrophy, fever, and mucopurulent ocular discharge. Radiographs typically reveal periosteal proliferation of various bones. Extreme leukocytosis is the most common laboratory finding, along with a mild elevation of serum alkaline phosphatase. Diagnosis is made by visualization of gamont-containing neutrophils or monocytes on examination of blood smears; observation of typical cysts, meronts or pyogranulomas on muscle biopsy; or detection of serum antibodies against H americanum sporozoites. Common complications of chronic infection include glomerulopathies, amyloidosis, and vasculitis. Although the prognosis for this disease in the past was guarded to poor, recent advances in treatment have increased the long-term survival rate of infected dogs. PMID- 12910750 TI - Canine visceral leishmaniasis and its emergence in the United States. AB - Leishmaniasis is an important zoonotic disease that accounts for approximately 57,000 deaths among people each year. Dogs are reservoirs for the disease, and dog ownership is a risk factor in endemic areas. This article discusses insect vectors for this disease, as well as diagnosis and treatment for dogs. PMID- 12910751 TI - Rabies among infrequently reported mammalian carnivores in the United States, 1960-2000. AB - Most cases of rabies reported annually in the United States occur among three groups of carnivores--raccoons (Procyon lotor), skunks (Mephitis, Spilogale, and Putorius), foxes (Vulpes, Urocyon, and Alopex)--and among bats (numerous species). However, between 1960 and 2000, a total of 2,851 cases of rabies in 17 other carnivore taxa were reported to the Centers for Disease Control and Prevention, Atlanta, Georgia (USA), from 49 states, the District of Columbia, and Puerto Rico. Three species of these other carnivores (mongooses [Herpestes javanicus], coyotes [Canis latrans], and bobcats [Lynx rufus]) accounted for 92% (2,624/2,851) of the cases reported among other canivorous mammals (OCMs). Most OCMs demonstrated temporal or spatial variation in numbers of reported cases. Tests of specimens from OCMs infected in the United States identified variants of the rabies virus that corresponded with variants associated with the major terrestrial reservoirs within their respective regions of origin. Variants of the rabies virus in samples from mongooses in Puerto Rico could not be distinguished from those in samples from dogs in Puerto Rico by virus typing methods. PMID- 12910752 TI - Rabies emergence among foxes in Turkey. AB - Sixteen rabies isolates recently collected from mainland Turkey and two isolates held within a British archive were used to form a representative cohort from a range of vectors, and were analyzed to identify potential causes for an increase of rabies within the fox (Vulpes vulpes) population in Turkey. Each isolate was characterized by sequence analysis of the nucleoprotein gene and compared phylogenetically to the cohort, to isolates from neighboring countries and to isolates from continental Europe and Russia. From this analysis the isolates could be divided into three groups associated with geographic location. This included a western group, an eastern group, and one isolate that did not group with any other Turkish isolate. This observation was also found using the heteroduplex mobility assay as an alternative method for typing rabies virus isolates. Further comparison with isolates from neighboring countries suggests that this isolate was related to viruses present in Georgia and could represent a recent import to Turkey from that country. Within the two larger groups, sequence data were obtained from both infected dogs and foxes suggesting that there has been transmission of virus between these two species. The direction of transmission could not be identified by the phylogenetic analysis, although absence of rabies within the fox population in previous years suggests that this could represent a recent spillover from the domestic dog to the fox. PMID- 12910753 TI - Rapid and simple method for screening wild rodents for antibodies to Sin Nombre hantavirus. AB - Sin Nombre hantavirus (SNV) is the primary etiologic agent of hantavirus cardiopulmonary syndrome (HCPS) in the United States and Canada. Hantavirus cardiopulmonary syndrome is a zoonotic disease. The most common reservoir is the deer mouse (Peromyscus maniculatus), although numerous other species of wild rodent can carry the viruses that cause HCPS throughout the Americas. Infected rodents show no signs of clinical disease but they develop persistent infection. Sin Nombre virus can be contracted by exposure to feces, urine, or saliva of its rodent reservoirs. Detection of infection in rodents is most often based upon detection of specific antibodies; many laboratories use enzyme linked immunosorbent assays (ELISAs), which require a specialized electrical ELISA reader. Enzyme linked immunosorbent assay readers are not readily amenable to field usage. We describe a portable test, the strip immunoblot assay (SIA), which we have utilized in field diagnosis. The test can be conducted in approximately 6 hr during the day or can be conducted overnight. The test can be used to detect rodents positive for SNV antibody while they are in traps. We show that results with the SIA have excellent concordance with western blot and reverse transcriptase polymerase chain reaction tests. PMID- 12910754 TI - Modeling control of rabies outbreaks in red fox populations to evaluate culling, vaccination, and vaccination combined with fertility control. AB - A predictive model of spread and control of rabies in red fox (Vulpes vulpes) populations was used to evaluate efficacy of culling, oral vaccination, and oral vaccination and fertility control (V + FC) as rabies control strategies. In addition, effects of season, fox population density, and a delay in starting control were modeled. At fox densities of 0.5 fox families/km2 or greater, a single oral vaccination campaign with bait uptake rates of less than 50% resulted in ineffective rabies control. An uptake rate of at least 80% was required to give a better than 80% chance of eliminating rabies. Vaccination was least effective at controlling rabies if applied 1 or 2 mo before the foxes gave birth. Seasonal timing of poison or V + FC had little effect on efficacy, which was always more successful than the oral vaccination alone. The longer the delay between the simulated start of the rabies infection and the application of a single vaccination campaign, the less successful was the control, particularly at the higher fox densities tested. At a fox density of 0.25 families/km2, all the strategies were equally successful at eliminating rabies. At higher fox densities V + FC was slightly less successful than culling, whereas vaccination-only was considerably less successful. The sole use of vaccination is not considered a viable control method for areas with high fox densities. The model suggests that an area of culling centered on the disease focus, plus an outer ring of vaccine or V + FC, could be the best strategy to control a point-source wildlife rabies outbreak. PMID- 12910755 TI - Analyses of gene flow among populations of deer mice (Peromyscus maniculatus) at sites near hantavirus pulmonary syndrome case-patient residences. AB - Gene flow and potential for Sin Nombre virus (SNV) trafficking of the deer mouse (Peromyscus maniculatus) was studied in Delta and Mesa counties of western Colorado (USA). The study areas included Grand Mesa and surrounding grazing and agricultural areas. This area has several natural potential barriers to rodent gene flow, including rivers, cliffs, and mountains. Ten study sites were utilized in a spatially nested design ranging from 0.65-81 km apart; four of these sites were at or near human hantavirus pulmonary syndrome (HPS) case-patient residences. One HPS case occurred on the north side of Grand Mesa in 1993; the other three (two confirmed, one presumptive) occurred on the south side of Grand Mesa between 1999-2000. Blood and tissue samples were collected from each of 221 deer mice captured from 1999-2000. Blood samples were tested for IgG antibody to SNV. At least one deer mouse had antibody to SNV at nine of 10 sites. Genomic DNA was isolated from tissue samples and alleles at six microsatellite loci were amplified by polymerase chain reaction (PCR). Polymorphisms were resolved on denaturing polyacrylamide gels and visualized by silver staining. Traditional population genetic analyses of this study indicated moderate population subdivision among the populations surveyed, slight evidence of isolation by distance, and that the Gunnison River system may hinder gene flow in this area. Application of assignment tests indicated that approximately 73-85% of mice were assigned to their population of capture. Many of the misclassifications occurred among sites less than 1 km apart; however, some long-distance misclassifications were noted. Additionally, some misclassifications were noted among study sites on different sides of the Gunnison River system, indicating that the riparian corridor of this system may facilitate some gene flow. Overall, these data indicate that SNV trafficking is more likely at the local level, but some long distance trafficking may be possible, especially where select habitat variables favor long-distance movements. PMID- 12910756 TI - Hantavirus antibody occurrence in bank voles (Clethrionomys glareolus) during a vole population cycle. AB - Puumala virus, genus Hantavirus, is the etiologic agent of nephropathia epidemica, a mild form of hemorrhagic fever with renal syndrome. The bank vole (Clethrionomys glareolus) is the natural reservoir species of this hantavirus. We initiated sampling of bank voles at sites of recently identified human nephropathia epidemica cases and paired control sites in the fall of 1995 in coastal areas of northern Sweden. Sites were trapped annually in spring and fall until 1999. Prevalence of antibody to Puumala virus was similar among local bank vole populations in the two types of sites over time. During peak years, however, the absolute number of bank voles was higher in case sites than control sites. Consequently, the likelihood of Puumala virus exposure was increased at case sites during population highs. This would imply that the risk of Puumala virus exposure to conspecifics and humans is habitat and site dependent with a temporal component. PMID- 12910757 TI - Mortality of common loons in New England, 1987 to 2000. AB - Diagnostic findings are presented on 522 common loons (Gavia immer) found dead or moribund in New England (Connecticut, Maine, Massachusetts, New Hampshire, Rhode Island, and Vermont, USA) between 1987 and 2000. Common loon numbers and range in New England have decreased from historic levels over the last century due to a number of proposed factors. Goals of this study were to identify and categorize causes of mortality and quantify natural versus anthropogenic causes. The majority of identifiable mortality in chicks was from intraspecific aggression (25%) and other causes of trauma (32%). Death in immature loons was primarily from fungal respiratory disease (20%) and trauma (18%). Causes of adult loon mortality differed significantly in breeding and wintering habitats. Wintering adults primarily died of trauma (17%) and infection (11%) and had significantly poorer body condition than breeding loons. In breeding adults, confirmed and suspected lead toxicosis from ingested fishing weights accounted for almost half of all mortality. Direct anthropogenic factors accounted for 52% of loon mortality in this study. Because of high carcass recovery rates, we believe these data are a good representation of loon mortality in New England. Results highlight the importance of human influences on conservation and management of the common loon in New England. PMID- 12910759 TI - Testing for Mycobacterium avium subsp. paratuberculosis infection in asymptomatic free-ranging tule elk from an infected herd. AB - Forty-five adult tule elk (Cervus elaphus nannodes) in good physical condition were translocated from a population located at Point Reyes National Seashore, Marin County (California, USA), to a holding pen 6 mo prior to release in an unfenced region of the park. Because infection with Mycobacterium avium subsp. paratuberculosis (Mptb) had been reported in the source population, the translocated elk underwent extensive ante-mortem testing using three Johne's disease assays: enzyme linked immunosorbent assay (ELISA); agar gel immunodiffusion assay (AGID), and fecal culture. Isolation of Mptb was made from fecal samples in six of 45 elk (13%). All AGID results were negative while ELISA results for 18 elk (40%) were considered elevated. Elevated ELISA results or Mptb isolation from fecal samples were obtained for 22 of 45 elk (49%); these elk were euthanized and necropsied. Mycobacterium avium subsp. paratuberculosis was isolated from tissue in 10 of 22 euthanized elk (45%); of these 10 cases of confirmed infection, eight had elevated ELISA results (80%) and four were fecal culture positive (40%). One of 10 cases had histopathologic lesions consistent with Mptb infection. Mycobacterium avium subsp. paratuberculosis was also isolated from tissue from one of eight fetuses sampled. The number of tule elk found to be infected was unexpected, both because of the continued overall health of the source herd and the normal clinical status of all study animals. PMID- 12910758 TI - Validation of a Brucella abortus competitive enzyme-linked immunosorbent assay for use in Rocky Mountain elk (Cervus elaphus nelsoni). AB - Brucellosis caused by infection with Brucella abortus is present in some elk (Cervus elaphus nelsoni) of the Greater Yellowstone Area (parts of Wyoming, Montana, and Idaho, USA). Since 1985, the Wyoming Game and Fish Department has vaccinated elk on elk feedgrounds in northwestern Wyoming during the winter months using B. abortus strain 19 (strain 19). Analysis of this vaccination program is hampered by the inability of standard serologic tests to differentiate between strain 19 vaccinated elk and those exposed to field strain B. abortus. In 1993, a competitive enzyme-linked immunosorbent assay (cELISA) was licensed to serologically differentiate between strain 19 vaccinated cattle and cattle exposed to field strain B. abortus. Seven groups of elk sera representing various B. abortus exposure histories were used to validate the cELISA test for elk. The cELISA test differentiated strain 19 vaccinated elk from elk that were challenged with B. abortus strain 2308, a pathogenic laboratory strain. The specificity of the cELISA was 96.8% for elk vaccinated with strain 19 only and sampled between 6 mo and 2 yr post vaccination, or with no B. abortus exposure. The sensitivity of the cELISA was 100%. The cELISA test will be useful in evaluating sera collected from elk in vaccinated, brucellosis endemic herds in the Greater Yellowstone Area. PMID- 12910761 TI - Distribution and ecology of meningeal worm, Parelaphostrongylus tenuis (Nematoda), in northcentral North America. AB - Meningeal worm (Parelaphostrongylus tenuis), a common nematode parasite in white tailed deer (Odocoileus virginianus) and pathogenic for several species of ungulates in eastern North America, is not known to occur in the west. Heads of 1,902 white-tailed deer were examined for adult meningeal worm to determine geographic distribution of the parasite in Saskatchewan and Manitoba (Canada) and North Dakota (USA). Finding the parasite in a deer in eastern Saskatchewan near the Manitoba border established the current northern and western limits in Canada. Prevalence of infection was < 1, 18.6, and 8.2% in Saskatchewan, Manitoba, and North Dakota, respectively. Infected deer occurred throughout southern Manitoba and eastern North Dakota. Distribution appears to have changed little since the last published survey for P. tenuis in the region in 1972. We examined precipitation, temperature, deer density, and forest cover as likely correlates to prevalence and distribution of P. tenuis. Deer management units used for hunting purposes were the scale of analysis in the three jurisdictions. Presence of P. tenuis was positively correlated with precipitation during frost free periods and deer density, and it was negatively correlated with winter and spring temperatures. Landscapes with > 25 and < 75% forest cover were most likely to have infected deer. Low rainfall and low density of white-tailed deer likely influence the westernmost limit of P. tenuis. PMID- 12910760 TI - An outbreak of fungal dermatitis and stomatitis in a free-ranging population of pigmy rattlesnakes (Sistrurus miliarius barbouri) in Florida. AB - Between September 1997 and March 1998, a severe skin, eye, and mouth disease was observed in a population of dusky pigmy rattlesnakes (Sistrurus miliarius barbouri), at the Lake Woodruff National Wildlife Refuge in Volusia County, Florida (USA). Three affected pigmy rattlesnakes were submitted for necropsy. All snakes had severe necrotizing and predominantly granulomatous dermatitis, stomatitis, and ophthalmitis, with involvement of the subadjacent musculature and other soft tissues. Numerous fungal hyphae were seen throughout tissue sections stained with periodic acid Schiff and Gomori's methenamine silver. Samples of lesions were cultured for bacteria and fungi. Based on hyphae and spore characteristics, four species of fungi were identified from culture: Sporothrix schenckii, Pestalotia pezizoides, Geotrichum candidum (Galactomyces geotrichum), and Paecilomyces sp. While no additional severely affected pigmy rattlesnakes were seen at the study site, a garter snake (Thamnophis sirtalis) and a ribbon snake (Thamnophis sauritis) with similar lesions were found. In 1998 and 1999, 42 pigmy rattlesnakes with multifocal minimal to moderate subcutaneous masses were seen at the study site. Masses from six of these snakes were biopsied in the field. Hyphae morphologically similar to those seen in the severe cases were observed with fungal stains. Analysis of a database representing 10,727 captures in previous years was performed after the 1998 outbreak was recognized. From this analysis we determined that 59 snakes with clinical signs similar to those seen during the 1998 outbreak were documented between 1992 and 1997. This study represents the first documented report of a mycotic disease of free-ranging snakes. PMID- 12910762 TI - Survey for coccidia and haemosporidia in the lesser prairie-chicken (Tympanuchus pallidicinctus) from New Mexico with description of a new Eimeria species. AB - Blood films and fecal samples of the lesser prairie-chicken (Tympanuchus pallidicinctus) were examined for parasites when we surveyed specimens captured during a radio-tracking study conducted in Chaves County, New Mexico (USA). All birds were captured on the Caprock Wildlife Habitat Management Area, administered by the Bureau of Land Management. Samples were collected in late March, April, and early May 1998-2000. Oocysts were detected in five of 64 (8%) birds sampled and, upon sporulation, were determined to be an Eimeria species. This is the first eimerian reported from the lesser prairie-chicken and is described here as a new species. Sporulated oocysts are ellipsoidal, 27.1 x 22.7 (22-32 x 18-26) microns, with micropyle absent, but oocyst residuum and polar granule present. Sporocysts are ovoidal, 11.9 x 7.8 (10-14 x 6-10); a Stieda body, and sporocyst residuum are present, as is a small, indistinct substieda body. Inspection of blood smears revealed four cases of Plasmodium infection of 32 (13%) individuals sampled. The characteristics of this plasmodiid are consistent with the description of Plasmodium (Giovannolaia) pedioecetii, previously found in T. pallidicinctus (Stabler, 1978). PMID- 12910763 TI - Standard sampling techniques underestimate prevalence of avian hematozoa in willow ptarmigan (Lagopus lagopus). AB - A total of 68 willow ptarmigan (Lagopus lagopus L.) was collected during September 1995 from two localities in Troms County, northern Norway. Thin blood smears were prepared and examined for blood parasites. Of the 68 willow ptarmigan examined, 94% harbored one or more species of hematozoa. There were four (6%), 44 (65%), 16 (24%), and four (6%) birds infected by zero, one, two, and three species of parasites, respectively. Prevalences at the coastal locality, Kattfjord (n = 43), were Leucocytozoon lovati 86%, Trypanosoma avium (26%), and microfilariae (30%). At the inland locality, Iselvdalen (n = 25), prevalences were L. lovati 96%, T. avium 12%, and microfilariae 0%. We also searched connective tissues for the filaroid nematode Splendidofilaria papillocerca; in Kattfjord this parasite only occurred in adult hosts where prevalence was 94%, but the parasite was not found in Iselvdalen. To estimate the efficiency of parasite detection by standard blood sampling techniques, we sampled peripheral blood from the brachial wing vein and blood from the pulmonary system from willow ptarmigan. Sampling peripheral blood from the brachial vein led to underestimates of the prevalence of microfilariae. There was no significant difference between L. lovati and T. avium prevalence in blood collected from the brachial vein or deep circulation. Age of host had a strong impact on prevalence, especially for S. papillocerca and microfilariae. PMID- 12910764 TI - Hematology, plasma biochemistry, and serosurvey for selected infectious agents in southern giant petrels from Patagonia, Argentina. AB - In conjunction with reproductive and feeding ecology studies on southern giant petrels (SGP, Macronectes giganteus) blood samples were collected for baseline health evaluations. Twenty-five adult SGP from a breeding colony in Chubut, Argentina, were sampled during two consecutive breeding seasons, 1999-2000 (n = 15) and 2000-01 (n = 10). Values for hematology, plasma biochemistry, and minerals are described for 20 birds in apparent good physical condition. A serologic survey of exposure to selected infectious agents was also conducted on all 25 birds sampled. Southern giant petrels were serologically negative for evidence of exposure to infectious laryngotracheitis virus, avian encephalomyelitis virus, avian influenza virus, avian reovirus, infectious bursal disease virus, infectious bronchitis virus, paramyxovirus 1, 2, and 3 virus, Chlamydophila, and Aspergillus. Antibodies to avian adenovirus were found in 14% of SGP during the first sampling season, and 60% in the second year. Additionally, all birds were negative for antibodies to Salmonella pullorum at the first sampling date, but 90% had low titers the following breeding season. This study contributes to understanding the health status of South Atlantic seabirds and to establishment of baseline information for SGP. Long-term monitoring of pelagic predator-scavenger seabirds such as SGP should be established for the surveillance of marine ecosystem health. PMID- 12910765 TI - Effects of acepromazine on capture stress in roe deer (Capreolus capreolus). AB - The aim of this study was to evaluate effect of a short-acting neuroleptic (acepromazine) on capture stress response in roe deer (Capreolus capreolus). Sixteen roe deer were captured by drive-nets in the winters of 1998, 1999, and 2001. Roe deer were divided into two groups: animals in the treatment group received an intramuscular injection of acepromazine (0.093 mg/kg +/- 0.003 SEM; n = 8) while animals in the control group (n = 8) did not receive tranquilizer. Heart rate and body temperature, as well as hematologic and biochemical indicators of stress, were used to evaluate effect of the neuroleptic over 3 hr. Heart rate decreased over time after capture in both groups (P < 0.05), but stabilized sooner in the treated roe deer (75 min after capture) than in the controls (105 min after capture). Body temperature decreased over 45 min and then stabilized in both groups (P < 0.05). Comparisons of blood parameters revealed significantly lower red blood cell count (RBC), lymphocyte count, hemoglobin concentration, packed cell volume (PCV), and serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), creatine kinase (CK), and lactate dehydrogenase (LDH) activities in tranquilized animals compared with controls (at least P < 0.05). A reduction in PCV, lymphocyte count, and serum cortisol concentrations (at least P < 0.05) and an increase in serum creatinine levels (P < 0.05) were recorded over time in control animals, while a reduction in RBC and hemoglobin concentration (at least P < 0.05) and an increase in serum urea concentrations (P < 0.05) over time were observed in the treated group. Finally, a decrease in serum lactate and potassium levels and an increase in CK, AST, ALT, and LDH activities were recorded over time in both groups. Results obtained showed the suitability of using acepromazine in capture operations in order to reduce stress response and prevent its adverse effects in roe deer. The beneficial effect was not only due to the sedative effect of acepromazine, but also to peripheral vasodilatation. PMID- 12910766 TI - Comparison of isoflurane and sevoflurane for anesthesia in beaver. AB - We compared the hemodynamic and respiratory effects, recovery time, and cost of two gas inhalants (isoflurane and sevoflurane) for anesthetic induction and maintenance of beaver (Castor canadensis) during surgery to implant radio transmitters in the peritoneal cavity. Heart rate, respiratory rate, relative hemoglobin saturation with oxygen (SpO2), and body temperature were measured every 5 min for the first 45 min, and arterial blood gas was measured once, 25 min into the anesthetic procedure. Induction for either agent was smooth and rapid. Heart rate and respiratory rate both decreased during the procedure though neither was lower than baseline values reported in the literature for beaver. Relative hemoglobin saturation with oxygen, body temperature, and blood gas variables did not differ between each anesthetic regime. Both inhalants caused slight respiratory acidosis. Recovery time from anesthesia was highly variable (1 178 min) but did not differ statistically between drugs. Sevoflurane costs ($22.30/60 min) were much higher than isoflurane costs ($3.50/60 min). We recommend isoflurane or sevoflurane for anesthetic induction and maintenance of beaver because of the lack of physiologic differences. PMID- 12910767 TI - Immobilization of free-ranging European mink (Mustela lutreola) an polecat (Mustela putorius) with medetomidine-ketamine and reversal by atipamezole. AB - From March 1996 to August 1999, 24 free-ranging European mink (Mustela lutreola) and 25 free-ranging polecats (Mustela putorius) were immobilized for clinical procedures and to place radio transmitters. Data were recorded during 14 and 12 trials, respectively. Animals received intramuscularly 10 mg/kg ketamine (KET) combined with 0.20 mg/kg medetomidine (MED), antagonized by 1.00 mg/kg atipamezole (ATI). Anesthesia times were similar between species. Induction was smooth and rapid (0.7-3.9 min); the degree of anesthesia and muscle relaxation was satisfactory in most animals. Two individuals showed signs of spontaneous recovery before injection of ATI. In other individuals, ATI was injected 28.1 54.0 min after the MED-KET injection and rapidly reversed the effects of the MED. Rectal temperature and heart and respiratory rates decreased significantly 5-25 min post MED-KET injection in both species. Rectal temperature successfully remained stable by placing animals on a warmed plastic table (37 C) during anesthesia. According to these results, this anesthetic protocol produces a safe and rapid immobilization in free-ranging European mink and polecats and is recommended for surgical procedures such as radio transmitter implantation. However caution is required as hypothermia can be severe. Body temperature must be monitored and means provided to maintain stability. PMID- 12910768 TI - Experimental vacuolar myelinopathy in red-tailed hawks. AB - Avian vacuolar myelinopathy (AVM) was recognized in 1994 as a cause of wild bird mortality when 29 bald eagles (Haliaeetus leucocephalus) succumbed to the disease at DeGray Lake, Arkansas (USA). The cause of AVM and its source remain undetermined despite extensive diagnostic and research investigations. Two years later, when AVM killed 26 eagles in the same area in Arkansas, it became apparent that American coots (Fulica americana) had identical neurologic signs and lesions, and it was hypothesized that eagles acquired AVM via ingestion of affected coots. In order to test this hypothesis, we fed coot tissues (brain, liver, kidney, muscle, fat, and intestinal tract) to rehabilitated, non releasable red-tailed hawks (Buteo jamaicensis). Five hawks received tissues from coots with AVM lesions, and one hawk received tissues from coots without brain lesions that had been collected at a site where AVM never has been documented. All hawks received 12-70 g/day (mean = 38 g) of coot tissues for 28 days. All six hawks remained clinically normal during the study. The birds were euthanatized on day 29 and microscopic lesions of AVM were found in all hawks that received tissues from affected coots, but not in the hawk that received tissues from unaffected coots. This marks the first time that AVM has been produced in birds under laboratory conditions and proves that birds of prey can acquire AVM via ingestion of tissues from affected coots. PMID- 12910769 TI - Lawsonia intracellularis in wild mammals in the Slovak Carpathians. AB - Feces of wild mammals were collected in the Bukovske Vrchy Hills (north-eastern Slovakia) in January and February 2002. The feces were examined for Lawsonia intracellularis by means of nested polymerase chain reaction. A total of 194 samples of feces from red deer (Cervus elaphus), 46 samples from roe deer (Capreolus capreolus), 31 samples from red fox (Vulpes vulpes), 23 samples from gray wolf (Canis lupus), and 12 samples from brown hare (Lepus europaeus) were examined. Lawsonia intracellularis was found in two samples from wolves, in two samples from foxes, and one sample from red deer. This is the first description of L. intracellularis in these three species. PMID- 12910770 TI - Vacuolar myelinopathy in waterfowl from a North Carolina impoundment. AB - Vacuolar myelinopathy was confirmed by light and electron microscopic examination of mallards (Anas platyrhynchos), ring-necked ducks (Aythya collaris), and buffleheads (Bucephala albeola) collected during an epizootic at Lake Surf in central North Carolina (USA) between November 1998 and February 1999. Clinical signs of affected birds were consistent with central nervous system impairment of motor function (incoordination, abnormal movement and posture, weakness, paralysis). This is the first report of this disease in wild waterfowl (Anseriformes). PMID- 12910771 TI - Experimental aerosol inoculation of Mycobacterium bovis in North American opossums (Didelphis virginiana). AB - The goal of this study was to evaluate the susceptibility of North American opossums (Didelphis virginiana) to aerosol inoculation of Mycobacterium bovis at two dose levels in order to gain information on disease pathogenesis, fecal shedding of the organism, and the potential role that opossums play in the spread of this disease in nature. Six opossums received high dose (1 x 10(7) colony forming units (cfu) by aerosol inoculation, six opossums received low dose (1 x 10(3) cfu inoculation, and six opossums were sham-inoculated with sterile water and served as controls. Lungs were the most frequently infected tissues, with nine of 12 inoculated opossums positive for M. bovis on culture. Gross lesions consisted of multifocal pneumonia and enlarged lymph nodes. Microscopically, granulomatous pneumonia and granulomatous lymphadenitis associated with acid-fast bacilli were present in eight of 12 inoculated opossums. Fecal shedding of M. bovis was uncommon at both inoculation doses. While opossums were highly susceptible to aerosol inoculation of M. bovis, they did not become emaciated or develop widely disseminated lesions. From this study, opossums may transmit tuberculosis by aerosol infection to other opossums in close contact and serve as a source of infection to carnivores that feed upon them, however, transmission of the disease to large herbivores by fecal shedding or direct contact may be less likely. PMID- 12910773 TI - Degenerative encephalopathy in a coastal mountain kingsnake (Lampropeltis zonata multifasciata) due to adenoviral-like infection. AB - In March 2000, an approximately 30-yr-old, male coastal mountain kingsnake (Lampropeltis zonata multifasciata) presented with disequilibrium and unresponsiveness to stimuli that ultimately lead to euthanasia. Histologically, there were foci of gliosis primarily within the caudal cerebrum, brainstem, and cervical spinal cord. Several glial cells and endothelial cells contained magenta, intranuclear inclusion bodies. Electron microscopy of the inclusions revealed paracrystalline arrays of 79-82 nm, viral-like particles. DNA in situ hybridization of sections of formalin-fixed brain using a mixture of two digoxigenin-end-labeled, adenovirus specific, oligonucleotide probes at low and high stringency was positive for adenovirus. PMID- 12910772 TI - An intraerythrocytic small piroplasm in wild-caught Pallas's cats (Otocolobus manul) from Mongolia. AB - During the quarantine examination of four Pallas's cats (Otocolobus manul) imported from Mongolia in October and December 2000, intraerythrocytic piroplasms were detected on Wright-Giemsa stained blood films that were morphologically indistinguishable from other small piroplasms of felids. Further characterization of this unknown organism via polymerase chain reaction amplification, sequencing of a portion of the 18S nuclear small subunit rRNA gene, and comparisons with orthologous sequences from other piroplasms, revealed similarity to Cytauxzoon felis. This is the first report of naturally occurring erythroparasitemia in Pallas's cats and the first documented case of naturally occurring piroplasm infections in a free-ranging felid from Mongolia. PMID- 12910774 TI - Chorioptic mange in a wild Japanese serow. AB - Parasitologic and histopathologic examinations were performed on a wild Japanese serow (Capricornis crispus) with severe dermatitis from Tochigi, Japan in February 1999. The serow was infested with a large number Chorioptes bovis. Marked parakeratotic hyperkeratosis was characteristic of the lesions induced by these mites. Japanese serow is a new host of C. bovis. PMID- 12910775 TI - Natural infection of Cryptosporidium muris (Apicomplexa: Cryptosporiidae) in Siberian chipmunks. AB - Coprologic examination of nine Siberian chipmunks (Eutamias sibiricus) imported from Southeast Asia revealed infection with Cryptosporidium sp. Experimental inoculation of BALB/c mice proved their susceptibility to the infection. Infected mice shed oocysts 14-35 days postinfection. Oocyst morphology was similar to that reported for C. muris in previous studies, oocysts were 8.1 (7.0-9.0) x 5.9 (5.0 6.5) microns. Clinical signs were absent in naturally infected chipmunks and experimental mice. Histologic examinations of mice revealed numerous developmental stages of C. muris in the glandular stomach. Analysis of partial small subunit rRNA gene sequences confirmed identity of these isolates as C. muris. Our results represent the first report of C. muris in members of the family Sciuridae. PMID- 12910776 TI - Paucity of hematozoa in Colombian birds. AB - Sixty-four birds of 43 species were caught at six localities in Colombia during the dry season in March 1998 and investigated for hematozoa by microscopic examination of stained blood films. Haemoproteus coatneyi, Plasmodium vaughani, Leucocytozoon sp., and microfilariae were identified. The overall prevalence of infection was 8%. Prevalences of infection for Haemoproteus spp., Plasmodium spp., Leucocytozoon spp., and microfilariae were 3%, 2%, 2%, and 3%, respectively. All hemosporidian infections encountered were of low intensity (< 1% of infected erythrocytes). The low prevalences and intensities of hemosporidian parasites in this study are in accord with other records from the Neotropics. PMID- 12910777 TI - Serologic survey for diseases in free-ranging coyotes (Canis latrans) from two ecologically distinct areas of Utah. AB - The influence of habitat and associated prey assemblages on the prevalence of canine diseases in coyotes (Canis latrans) has received scant attention. From December 1997 through December 1999, we captured 67 coyotes in two ecologically distinct areas of Utah (USA): Deseret Land and Livestock Ranch and US Army Dugway Proving Ground. These areas differ in habitat and prey base. We collected blood samples and tested for evidence of various canine diseases. Prevalence of antibodies against canine parvovirus (CPV) was 100% in the Deseret population and 93% in the Dugway population. All juveniles in both populations had been exposed. We found no difference in the prevalence of antibodies against canine distemper virus (CDV) between the two populations (7% versus 12%; P = 0.50). However, we did find an increase in antibodies with age in the Deseret population (P = 0.03). Evidence of exposure to canine adenovirus (CAV) was found in both populations (52% and 72%; P = 0.08). Prevalence of CAV antibodies was influenced by age on both areas (Deseret: P = 0.003; Dugway: P = 0.004). Antibodies to Francisella tularensis were low on both areas (2% and 4%). We found a significant difference (P = 0.001) in the prevalence of exposure to Yersinia pestis between the two populations: 73% in Deseret compared to 11% in Dugway. This difference is most likely due to the prey species available in the two ecologically distinct study areas. PMID- 12910779 TI - Hematology and serum chemistry of the island spotted skunk on Santa Cruz Island. AB - We determined serum biochemistry and hematologic values for island spotted skunks (Spilogale gracilis amphiala) on Santa Cruz Island (California, USA). Samples were collected from island spotted skunks chemically restrained with ketamine hydrochloride and acepromazine in August 1999 (dry season) and from skunks manually restrained in August 2000 (dry season) and January 2001 (wet season). One parameter, glucose, significantly differed with season, with higher levels during the wet season. Serum chemistry and hematologic profiles suggest that method of restraint (manual or chemical), as well as other methodologic details, may influence blood characteristics in the island spotted skunk. PMID- 12910778 TI - Ecologic correlates of Toxoplasma gondii exposure in free-ranging neotropical mammals. AB - A serologic survey for Toxoplasma gondii in 18 free-ranging forest mammal species (n = 456) in French Guiana was undertaken with a direct agglutination test. Serum antibody prevalence varied from 0-71%. The relationships between ecologic features of the species and seroprevalence were investigated. Terrestrial mammals were significantly more exposed to T. gondii than other mammals. This result is concordant with oral exposure to T. gondii related to ground dwelling behavior and/or carnivory. PMID- 12910780 TI - Morbidity and mortality of red foxes (Vulpes vulpes) and gray foxes (Urocyon cinereoargenteus) admitted to the Wildlife Center of Virginia, 1993-2001. AB - The medical records of 48 red foxes (Vulpes vulpes) and 35 gray foxes (Urocyon cinereoargenteus) examined at the Wildlife Center of Virginia (Waynesboro, Virginia, USA) from 1993 to 2001 were reviewed. The most common diagnosis in red foxes was orphaned (33%), followed by trauma (27%), undetermined diagnosis (23%), and sarcoptic mange (17%). Trauma (46%) was the most frequent cause of morbidity and mortality in gray foxes followed by orphaned (23%), undetermined (20%), toxoplasmosis (6%), presumptive canine distemper (3%), and rabies (3%). One gray fox had concurrent toxoplasmosis and presumptive canine distemper (3%). Similar diseases were detected in previous studies at a diagnostic laboratory; however in this study, trauma and orphaned animals were more common than infectious diseases. The lack of diagnostic information on some cases limited the usefulness of this study, and more emphasis should be placed on performing postmortem examinations of wildlife presented to wildlife rehabilitation centers. PMID- 12910781 TI - From leg transplantation by St Cosmas and St Damian to the modern era. PMID- 12910782 TI - The history of transplantation at the Catholic University of Louvain Belgium 1963 2003. PMID- 12910783 TI - Renal transplantation at K.U. Leuven. PMID- 12910784 TI - Organ transplantation pioneers at the "Universite Libre de Bruxelles". PMID- 12910785 TI - History of transplantation at the University Hospital of Ghent Belgium 1965-2002. PMID- 12910786 TI - History of solid organ transplantation at the University of Liege. PMID- 12910787 TI - History of transplantation at the University Hospital of Antwerp. PMID- 12910788 TI - William Welch and his personal history of organ transplantation at the Vrij Universiteit Van Brussel. PMID- 12910789 TI - Belgian pioneers in transplantation surgery. PMID- 12910802 TI - [HCV infection today]. PMID- 12910803 TI - [An effective method to prevent deep venous thrombosis in total hip arthroplasty: a study in patients with and without risk factors for thrombotic disease]. AB - PURPOSE: The aim of this study was to determine a safe and effective method of prophylaxis for thromboembolis diseases after THA. MATERIALS AND METHODS: This study was conducted on 157 patients consecutively admitted to our Department of Orthopedics to undergo elective THA from October 2000 o May 2001. We have preoperatively investigated plasma levels of homocysteine, AT III activity, Fibrinogen levels, Anticardiolipin antibodies, and circulating vWFag D-dimer levels were measured by Asserachrom D-dimer ELISA preoperatively and on day 4 postoperatively. Thromboprophylactic regimen was based on a prolonged subcutaneous administration of nadroparin (for 40 days after surgery) and was used in all patients, with a dose based on body weight. Compression ultrasonography was udes as screening test for the diagnosis of deep venous thrombosis and performed in each patient on postoperative day 4, 15, and 30. RESULTS: Although all patients enrolled in this study showed increased risk for thrombotic disease, none oh them developed either symptomatic or asymptomatic deep venous thrombosis. No complications were observed, including major bleeding or abnormalities of laboratory tests. CONCLUSIONS: Our study indicates that prolonged thromboprophylaxis with nadroparin for 40 days postoperatively, associated with early mobilization, is an effective and safe protocol of antithrombotic prophylaxis in patients operated for THA with and without risk factors for thrombotic disease. PMID- 12910804 TI - [Prevalence of HCV in health care workers in Southern Italy]. AB - PURPOSE: To assess the prevalence of hepatitis C virus (HCV) in the population exposed to biological risk of two hospitals in southern Italy. PATIENTS AND METHODS: All health care workers of two main hospitals of Sicily, potentially exposed to biological risk underwent health surveillance. Health care personnel were arranged into seven occupational group, and five age bands (< 30, 31-40 years, 25-30 years, 41-50, 51-60 years, > 60 years). We estimated the prevalence of hepatitis C infection in 1800 healthcare workers by means of nucleic acid amplification and genotype by means of sequencing, the workers were also tested for liver function. RESULTS: The overall prevalence of anti-HCV was 2.1%, corresponding to 39 persons. The prevalence in health workers was higher in nurses than in surgeon, group perceived to be at greatest risk of occupational exposure, and higher in elderly workers (> 45 years) than in younger ones. CONCLUSION: Prevalence of anti-HCV in our study was higher than in blood donors, and lower than in general population in southern of Italy. The 45% of HCV infected healthcare workers were unaware of their condition and their potential to infect patients. Testing for HCV infection should be routinely performed for health care workers to detect the infection, which frequently results in a chronic asymptomatic carrier state for many years before the development of symptomatic liver disease, and perform precocious therapy with interferon. PMID- 12910805 TI - [Laparoscopically assisted vaginal hysterectomy Gasless. Retrospective study in patients with uterine fibroids]. AB - PURPOSE: To evaluate the feasibility to perform laparoscopically assisted vaginal hysterectomy in patients who were not suitable for vaginal hysterectomy. PATIENTS AND METHODS: LAVH Gasless was carried out on 31 patients with uterine fibroids. Uterine weight ranged of 420-800 gr. RESULTS: There were 2 laparotomy conversion (6%) for uterus 800 gr; median operating time was 79 +/- 18 minutes (range 75 89); the average haemoglobin drop was 1.7 +/- 0.9 g/dl; the mean in postoperative stay was 3.7 +/- 0.7 days (range 3-5). Febrile morbidity > 38 degrees C were found in 3 cases. CONCLUSIONS: Laparoscopic assistance has advantages in performing transection of round and infundibolo-pelvic ligaments in vaginal steps and in controlling post-operative blood loss. PMID- 12910806 TI - [Experimental study on efficacy of thermal muds of Ischia Island combined with balneotherapy in the treatment of psoriasis vulgaris with plaques]. AB - A lot of studies show the efficacy of thermal therapy in psoriasis. The main indications concern all forms of psoriasis (except for pustolosa and eritrodermic) especially diffuse psoriasis, intensive itchy, artropatic psoriasis and gutted psoriasis child. Benefit effects of thermal water and muds, which are important parameters in psoriasis' treatment, they are excellent to reduce inflammation, scaling, and erythema. In this study we evaluated the clinical efficacy of thermal bath plus muds in patients affected by psoriasis. 18 patients has been examined and randomly divided in 3 group (A-B-C). All the patients received thermal therapy for 4 weeks. To evaluate the behaviour of the disease we performed (corneometry, sebometry, phmetry), as well as evaluation of P.A.S.I. At the end of the study, values of P.A.S.I. showed a reduction from 10 to 6 unites P.A.S.I. for patients belong to group A, more 12 to less 10 for patients belong to group C e no significant variations for group B (because represent control group). Results obtained can be considered useful, considering that thermal treatment was used alone in the treatment of all patients. PMID- 12910807 TI - [Counseling in Internal Medicine and Family Medicine: old and new needs of the patients]. AB - INTRODUCTION: Present proliferation of so called Complementary and Alternative Medicine (CAM), reveals the patient's deep discomfort, as s/he does no longer perceive as satisfying the relation with his/her family doctor, thus wanting to seek for different solutions. The lack of a scientific basis of the majority of these medicines often causes an effective risk to health. OBJECTIVE: The survey highlights how the acquisition of expertise in Counselling, together with the more specific clinical ones, enables the physician to manage the many relational issues more adequately. This skill will allow physicians to meet patients' needs more effectively, and as a consequence, it will make them more satisfied of the relation which will loose its present anonymous and bureaucratic nature. The intrinsic dignity of the help relation will be restored and this will help to face the treatment processes even if these do not bring to recovery. CONCLUSIONS: The new expertise proposed to physicians, could be part of the new ECM programmes. It could also allow to verify, in Italy as well, if a medical class more prepared under this aspect, corresponds to a reduction of request for CAM, and consequently to a substantial improvement of the scientific dimension of personal professional vocation. PMID- 12910808 TI - [Recent trends in the therapy of hepatitis C]. AB - The treatment of HCV correlated hepatitis is an important argument, because of the great incidence and prevalence of this disease. At the end of the Eighties, the IFN was the first substance used for HCV correlated hepatitis therapy. The IFN monotherapy with a dose of 3-6 MU for 6-12 months eradicates the infection in the 15% of cases, and cause an histological temporary improvement in a variable number of patients that keep the infection. In the following years, the research on evaluation of the efficacy of the recombinant interferons or interferons made with genetics engineering (IFN alpha 2a, IFN alpha 2b and IFN Consensus) has given results comparable with the results obtained with IFN alpha. Later, it started the experimentations with IFN associated to other substances, for example IFN-ribavirin combination therapy. The treatment IFN-Ribavirin eradicates the infection in 30% of the patients with the genotype 1b and in 60% with the genotype 2 or 3, while this treatment is less efficacious in the patients with the genotype 4. Recently, it started to use the PEG IFN. The pegylation is the combination of a polietylen-glicole molecule with the IFN molecule, so as to prolong its half-life and reduce the dose only one a week, with reduction of the collateral effects. Some studies has shown that the use of PEG-IFN in monotherapy could help the patients with advanced liver complaint. Successive studies are directed to show the efficacy of the PEG-IFN and ribavirin combination therapy. Recent researches put in evidence new substances, that could represent the future for HCV correlated hepatitis therapy. Between these substances we have to highlight the interleukin, the inhibitors of the viral multiplication and the inhibitors of IMPDH (Inosine monophosphate dehydrogenase). At the beginning of 2002 has made the improvement of HCV vaccine known. Actually, in the USA there are in progress human experimentations, and the production of gamma-globuline, that could be effective to prevent the infection. PMID- 12910809 TI - [Asthma and pregnancy]. AB - During pregnancy the asthma has an incidence of 1-4%. In addition to this pathology we find, during pregnancy, an increased incidence of complications both maternal and fetal. These complications, often serious, happen nearly exclusively in patients with severe asthma and/or badly controlled. Therefore, to reduce the risk of beginning of these complications it's necessary to assess correctly the gravity of asthma before becoming pregnant, to make a careful and periodic control of the respiratory function of the patient, to effect a suitable preventive and/or symptomatic pharmacological therapy and to establish specific sanitary-action measures able to prevent the disease becomes acute. PMID- 12910810 TI - [Management programs for elderly patients with chronic heart failure]. AB - PURPOSE: Most patients with heart failure are elderly with multiple coexisting diseases and heart failure is the most common discharge diagnosis in elderly hospitalized patients. Despite major advances in the pharmacotherapy of heart failure, hospitalization rates remain high, owing in large part to a multitude of psychosocial, behavioral, and financial factors that serve as barriers to effective compliance with prescribed treatment. In the last decade, several models have been proposed in order to optimise the long-term management of elderly patients with heart failure. DESIGN: A review of most significant and recent models available was performed. RESULTS: Several studies have documented the efficacy of specialized multidisciplinary heart failure disease management programs in terms of reducing hospital utilization, improving quality of life, functional capacity, patient satisfaction, compliance with diet and medications and decreasing cost of care. CONCLUSIONS: At present, the greatest challenge in managing elderly heart failure patients is to more effectively implement proven treatments and disease management systems. PMID- 12910811 TI - [Case report: Acro-necrosis of the upper limbs caused by gemcitabine therapy]. AB - INTRODUCTION: Even if infrequent, a digital necrosis after chemotherapy can occur in cancer patients. The gemcitabine is generally well tolerate; the cutaneous toxic ulcerations only in 0.3% of the cases induces the suspension of the treatment. CLINICAL CASE: A 70 year old patient, female, with a bladder cancer, after a trans-urethral resection, is submitted to adjuvant chemotherapy with Gemcitabine 1700 mg (total dose/die), with administration in the days 1st and 8th, while in the 15th day was not effected because, to distance of 3-4 days from the second administration, appear paresthesies of the fingers of the hands, together with Raynaud type phenomenon, 38-39 degrees C intermittent fever, digital necrosis and fingertips gangrene. Laboratory: (Normal): RF; AutoAb: AMA, ASMA, APCA, anti-DNA; ENA; lupus anti-coagulant; Ab-anti-cardiolipin; C3-C4, CIC; homocysteine, anti-thrombin, protein C, protein S, mutation of the factor V of Leiden, plasminogen, alfa 2-antiplasmin. (Altered): Auto-antibody: ANA (on Hep 2): positive (title 1/160, speckled pattern), cryoglobulin positive, ESR 29; Instrumental examinations: Superior Limbs Angiograpy: Occlusion of the digital arteries proper of 2nd, 3rd and 4th finger of the hands. Electromyography Inferior Arts: normal. Superior Arts: bilateral suffering of the median nerve at the carpal tunnel. Biopsy of the hand cutis: Hyperkeratosis, acanthosis and papillomatosis of the skin. Arterial vases with signs of endothelioangiitis and aspecific inflammation. CONCLUSIONS: Even if acronecrosis of the superior limbs is a rare effect of the gemcitabine, we would recommend particular caution in the administration of this drug in patient with known autoimmune disorders. PMID- 12910812 TI - [Guidelines for nutritional management in hospitals]. AB - Malnutrition, frequently seen in hospital patients, is associated with poor prognosis. Therefore rules of good care imply its early recognition and adequate treatment. Directions about nutritional and dietetic support aim at assuring the proper intake of calories and of other nutrients which are present in food. Guidelines to be used when prescribing a suitable diet for the hospitalized patient are presented. PMID- 12910814 TI - Biopsychosocial issues and risk factors in the family when the child has a chronic illness. AB - Between 10 and 20 million American children and adolescents have some type of chronic health condition or impairment. There has been a recent interest in the psychological aspects--emotional and behavioral--of chronic illness in children and adolescents. Major new areas of science, such as behavioral medicine, have emerged to address the research and clinical demands of this field. Several studies already have demonstrated the increased risk for psychiatric problems in children with chronic health conditions to be three to four times greater than their healthy peers. The proposed perspective shifts from a traditional sole reliance on medical diagnosis to a focus more on the dimensions of adjustment, socioeconomic status, visibility of condition, social support, and family functioning. PMID- 12910813 TI - [Colorectal polyps]. PMID- 12910815 TI - Child mental health consultation with families of medically compromised infants. AB - Prematurity and birth defects present parents with a crisis for which they have usually had little preparation and no prior education. Both types of early medical complications may represent a state of suspended animation for most parents. Even large premature infants with good prognoses induce anxiety and symbolize potential death and disability, and children with birth defects may portend years of medical procedures and long-term disability. The fear of serious neurologic impairment or mental retardation presents parents with a long period of ambiguity and chronic anxiety. During this period, they must be helpless observers rather than active participants. Recent research has indicated that the active involvement of parents in the care of their premature infants can be helpful in alleviating the guilt and anxiety related to loss and impairment. Similarly, early physical contact between parents and their severely malformed infant is equally critical. Even if the ultimate complexities of early attachment have yet to be delineated fully, this is a worthwhile practice and useful approach in the nursery. Child mental health professionals have important roles to fulfill in helping staff members deal with increased parental participation and directly managing family members with intense distress related to their infants' fragility. The role of the mental health professional in such consultation may cover five related tasks: 1. Understanding the nature of the biologic issues facing the child and integrating that understanding with an evaluation of the child's neurobehavioral profile. 2. Understanding the family's relationship with the child and their overall level of functioning during an acutely stressful time. 3. Developing an appreciation of the place of the child in his or her family and how the parents understand the nature of the medical problems. 4. Forming a collaborative relationship with the pediatricians and other subspecialists who care for the child so that behavioral and psychological interventions are integrated with the child's biomedical care. 5. Fostering a brief, or sometimes long-term, therapeutic relationship with the family or facilitating the family's finding such a relationship with another clinician. There will never be enough child and adolescent psychiatrists and psychologists to treat all families of medically compromised infants. Knowledge of normative responses has advanced to the point at which basic skills can be used by and transmitted to others who can provide basic services. There is much to be learned about the short- and long-term sequelae of such stressful situations on individuals and family systems with preexisting psychopathology. For such families, child mental health professionals are uniquely suited to play a further role in research and treatment. PMID- 12910816 TI - Psychological aspects of HIV infection in children. AB - Despite advances in the pharmaceutical treatment of HIV disease, there are still an increasing number of people living with the disease, and an increasing number of children and adolescents are personally affected by the epidemic. The psychological effects on these children are significant and relate not only to a parent's degree of illness and the threat of death but also to the association of the disease with substance abuse and the pervasive effects of the stigma that surround it. To intervene optimally on behalf of these children, programs must be multidisciplinary and take a holistic approach to address specific social and psychological issues and ensure stability in a child's care giving. PMID- 12910817 TI - Psychological aspects of diabetes mellitus. AB - Diabetes mellitus (DM) presents itself in two forms: insulin-dependent (type 1 DM) and non-insulin-dependent (type 2 DM). Although type 2 DM usually has an adult onset, in recent years there has been a significant rise in the number of children diagnosed with type 2 DM in the United States. Reasons for this increased frequency are believed to be a larger percentage of children who are overweight, a family history of diabetes, and a considerable increase in the use of psychotropic medication in children. The diagnosis of DM is a significant stressor not only for patients but also for their environment. Children with DM are sometimes stigmatized by their peers and relatives who do not understand the illness or are frightened by it. Some children also may need to alter several of their customary routines and are often scared to participate in activities in which they were previously engaged. The family's response to the diagnosis of DM may have a negative effect on glycemic control. Differences have been found in the way patients with type 1 DM and type 2 DM cope with and adapt to their diagnosis. PMID- 12910818 TI - Psychiatric symptoms in patients with asthma causality, comorbidity, or shared genetic etiology. AB - Despite the range of diverse studies that attempt to understand the comorbidity of asthma and psychiatric diagnoses, it is still not possible to provide a reliable quantitative estimate of the increased risk for anxiety and mood disorders in children with asthma. A new hypothesis for this comorbidity has evolved, however. It is likely that the stress of having a chronic illness increases the likelihood of the development of anxiety and depressive symptoms. If this were a sufficient etiologic explanation, however, increased comorbidities of psychiatric illnesses would be found in all chronic pediatric illnesses. More precise prevalence estimates of these comorbidities require the completion of large studies that use a longitudinal design and reliable and well-validated assessment instruments. The most promising direction for future research is the definition of underlying genetic vulnerabilities that reflect autonomic regulation, may contribute to the onset of some forms of asthma, and are associated with increased risk for anxiety and mood disorders. PMID- 12910819 TI - Psychiatric aspects of pediatric cancer. AB - Recent progress in the understanding of psychological and social factors related to cancer is important because cancer is the most common fatal disease of childhood and adolescence. Research interest in children and adolescents who have survived cancer also has increased over the past several years. Attention has focused on the long-term social outcomes of these children and adolescents. With increased survival, quality-of-life issues have assumed a more prominent role in the treatment protocols being developed for childhood cancer. The presence of long-lasting uncertainty about recurrence of the disease or second malignancy together with recognition of cognitive and physical side effects of treatment make childhood cancer a potentially chronic condition. Survivors experience actual or potential threats to future health; more than half have medical cognitive or psychological problems. PMID- 12910820 TI - Psychological aspects of traumatic injury in children and adolescents. AB - Each year millions of children are exposed to some form of extreme traumatic stressor. These traumatic events include natural disasters (e.g., tornadoes, floods, hurricanes), motor vehicle accidents, life-threatening illnesses and associated painful medical procedures (e.g., severe burns, cancer, limb amputations), physical abuse, sexual assault, witnessing domestic or community violence, kidnapping, and sudden death of a parent. During times of war, violent and nonviolent trauma (e.g., lack of fuel and food) may have terrible effects on children's adjustment. The events of September 11, 2001 and the unceasing suicidal attacks in the Middle East underscore the importance of understanding how children and adolescents react to disasters and terrorism. The body of literature related to children and their responses to disasters and trauma is growing. Mental health professionals are increasing their understanding about what factors are associated with increased risk (vulnerability) and affect how children cope with traumatic events. Researchers recognize that children's responses to major stress are similar to adults' (reexperiencing the event, avoidance, and arousal) and that these responses are not transient. A review of the literature indicates that PTSD is the most common psychiatric disorder after traumatic experiences, including physical injuries. There is also evidence for other comorbid conditions, including mood, anxiety, sleep, conduct, learning, and attention problems. In terms of providing treatment, CBT emerges as the best validated therapeutic approach for children and adolescents who experienced trauma-related symptoms, particularly symptoms associated with anxiety or mood disorders. The best approach to the injured child requires injury and pain assessment followed by specific interventions, such as pain management, brief consultation, and crisis intervention immediately after the specific traumatic event. Family support also may be necessary to help the family through this difficult period. The main conclusion that arises from the research on resilience in development is that extraordinary resilience and recovery power of children depend on basic human protective systems operating in their favor. This finding has produced a fundamental change in the framework for understanding and helping children at high risk or already in trouble. This shift is evident in a changing conceptualization of the goals of prevention and intervention that currently address competence and problems. Strategies for fostering resilience described in this article should be tested in future controlled psychotherapy trials to verify their efficacy on children's protective factors. PMID- 12910821 TI - Pediatric inflammatory bowel disease. AB - The treatment of children and families affected by pediatric IBD requires an awareness of the diverse psychobiologic effects of the disorder over the course of child, adolescent [53], and adult development [52]. Optimal treatment requires careful coordination of various medical, educational, and rehabilitative services and concerned, empathetic, continuity of care by knowledgeable clinicians [4]. In many cases, this care must also include individual psychotherapeutic work with affected youngsters at crucial junctures to help deal with the social and psychological stresses of IBD, as well supportive casework for parents and siblings. PMID- 12910822 TI - Psychiatric issues of childhood seizure disorders. AB - Children who suffer from childhood seizure disorders, especially epilepsy, have various potential psychiatric issues and concerns that the treating physician and psychiatric consultant should consider. These children are at increased risk of adjustment reactions, anxiety and mood disorders, ADHD, learning difficulties, and familial and social stress. Because of potential risks and vulnerabilities for the development of comorbid psychiatric conditions and the increased risk for individual, familial, and social impairment, a psychiatric consultation to children and families dealing with epilepsy may play an important role in the successful management of this complex disorder. PMID- 12910823 TI - Psychological aspects of hematologic diseases. AB - This article focused on two hematologic diseases that have received significant attention in the pediatric literature: sickle cell anemia and hemophilia. As the data indicate, these illnesses present lifelong challenges to individuals afflicted. With advances in medical interventions, many children with these hematologic diseases are living longer and with fewer serious complications. Intervention efforts only recently have begun to consider issues related to quality of life and increased psychological adjustment, including active coping strategies by children and their parents and necessary social support for all family members. Theoretical models [54] have highlighted the multiple factors that play a role in illness adjustment and the complex interactions of these factors. Individual, family, and community characteristics are impacted by and, in turn, impact on illness-related characteristics. The notion that illness severity is predictive of psychological adjustment has been discarded in favor of a model that recognizes multiple influences and multiple outcomes. Child coping, parent coping, social support, adaptive functioning, treatment compliance, and illness severity are being considered to better understand and influence overall psychological functioning. The field of pediatric psychology has made tremendous advances in improving knowledge of illness and its impact on development. Even with these advances, however, there remains much to be discovered. To date, pediatric psychology research has focused primarily on individual illness categories, making comparisons across illness types difficult. By considering different illnesses within the same study, characteristics that are crucial to improved adjustment and common across illnesses can be identified. As we continue to work for cures for these debilitating illnesses, our goal remains to improve quality of life for children. PMID- 12910824 TI - Psychosocial issues for children and adolescents with chronic illness: self esteem, school functioning and sports participation. AB - Self-esteem, school functioning, and sports participation are among the most significant psychosocial issues that affect children and adolescents with chronic illness. Although these capacities are essential components of development for all children, they present special concerns for children with limitations of health. Parents, teachers, and coaches play important roles in providing normalizing and gratifying opportunities for children who struggle to be competent and accepted by their peers. Much can be done to provide chronically ill children with experiences and support that will allow them to grow up happier, feel better about themselves, and enjoy more success. PMID- 12910825 TI - [Depressive symptoms following ischemic stroke]. AB - Depressed mood and other depressive symptoms frequently seen after cerebral stroke contribute to an unfavorable prognosis in this patient population. Identification of the subgroup of patients at increased risk for depressive symptoms is a prerequisite of early treatment. In the study aimed at evaluation of post-stroke depressive symptoms prevalence and risk factors participants were 766 consecutive patients with ischemic cerebral infarction, admitted in the years 1997-2000 to the Stroke Unit, Neurology Department in Cracow. Data concerning depressive symptoms, demographic characteristics and clinical variables were obtained from medical records. Depressive symptoms during the hospitalization were found in 19% of cases. Younger age, neurological deficits and previous history of psychiatric disorders were independent factors increasing the risk of post-stroke depressive symptoms. The study allowed to identify the subpopulation of stroke patients at risk for affective disorders following a cerebrovascular accident. PMID- 12910826 TI - [Prevalence of carotid arteries atherosclerotic changes in ischemic stroke patients]. AB - The aim of our study was to investigate prevalence, characteristics and extensiveness of CA plaques in acute stroke using high-resolution B-mode USG. METHODS: USG of CA was performed with a 7.0-MHz Duplex-type scanner Acuson 128 XP/10 C. We examined 149 patients aged 26-95 years, with acute ischemic stroke, consecutively admitted to our Department. Atheromatous plaques morphology was determined using the HSP Classification (H--Hemodynamic characteristics, S- Surface plaque, P--Plaque echogenics). RESULTS: An extracranial carotid disease (CCA, ICA, ECA) was found in 85 (57%) patients, while 67 (45%) patients had atheromatous changes in ICA. Bilateral ICA plaques were found in 22 (15%) patients, and unilateral--in 45 (30%) patients. In 17 (11%) patients plaques were associated with artery occlusion (in 16 cases ipsilateral to the stroke site, and in 1 case--contralateral). No bilateral occlusion of ICA was observed. In 6 (4%) patients stenosis (ranging from 60 to 99%) was found in only one ICA, while in 4 (2.7%) cases there was also contralateral stenosis < 60%. In 4 patients a contralateral occlusion was present. Ulcerated plaques were noted in 22 (15%) patients--in 18 cases ipsilateral to the stroke, and in the remaining 4- contralaterally. None of the patients had a bilateral ulceration. CONCLUSION: Atheromatous ICA changes were found in 45% of the patients under study. However, these plaques may be the cause of cerebral ischemia associated with a high-grade stenosis, occlusion or ulceration ipsilateral to the stroke site in 15.45% of the patients only. PMID- 12910827 TI - [Causes of excessive daytime sleepiness in hypercapnic and normocapnic obstructive sleep apnea patients]. AB - Daytime sleepiness assessed using the Epworth sleepiness scale (ESS) and polysomnography results were compared in 43 patients with obstructive sleep apnea (OSA) with concomitant chronic hypercapnia (PaCO2 53 +/- 6 mmHg), and in 58 patients with the OSA syndrome accompanied by normocapnia (PaCO2 < or = 45 mmHg, mean 39 +/- 3 mmHg). The OSA patients with hypercapnia were more sleepy than those with normocapnia (ESS 18 +/- 7 vs 15 +/- 7, p < 0.05), but apnea index values were similar in both groups (54 +/- 20 and 49 +/- 17). The following parameters of electrophysiological sleep structure were obtained in the hypercapnic OSA patients: sleep stage 1: 66 +/- 28%, stage 2: 28 +/- 27%, stage 3 + 4: 1 +/- 1%, REM sleep 5 + 6% of the total sleep time, while in the OSA patients with normocapnia: stage 1: 39 +/- 19%, stage 2: 28 +/- 27%, stage 3 + 4: 2 +/- 2%, and REM sleep 6 +/- 7% of the total sleep time. Stage 1 NREM sleep was found to be longer, and stage 2 NREM--shorter in hypercapnic than in normocapnic OSA patients (p < 0.01). CONCLUSION: Increased daytime sleepiness in both groups patients with the OSA syndrome is due to sleep fragmentation as well as to deficiency of deep and paradoxical sleep (almost absent deep sleep and extremely shortened REM sleep). Hypercapnic OSA patients' more marked sleepiness may result from a more pronounced disturbance of their sleep macrostructure, with a considerable predomination of stage I NREM sleep. PMID- 12910828 TI - [Prevalence of Helicobacter pylori infection among patients with multiple sclerosis]. AB - A group of 90 subjects with multiple sclerosis was examined with the serological Helicotest for the presence of antibodies against Helicobacter pylori. Positive results were found in 17 patients (18.9%), which is markedly lower than this infection incidence rate in the population of Poland. The results indicate a lack of connection between the Helicobacter pylori infection and multiple sclerosis. However, it seems reasonable to perform the test in MS patients so as to avoid the risk of the upper alimentary tract complications in the course of intense steroid treatment applied as a standard management of multiple sclerosis relapse. PMID- 12910829 TI - [Status epilepticus in the course of epilepsy in children and adolescents]. AB - Status epilepticus (SP) is defined as a single seizure or recurrent seizures of over 30 min. duration, without regaining full consciousness. In approximately 50% of cases SE is related to epilepsy. The International League Against Epilepsy has recently proposed a new SE classification (2001). The purpose of this prospective study is to evaluate SE incidence in children and adolescents in relation to their age and type of epilepsy, as well as to determine usefulness of the new classification of SE. A group of 600 children and adolescents with epilepsy was observed prospectively. The inclusion criteria were: recently diagnosed epilepsy in children aged under 15 years, and long-term treatment and observations (mean = 5 years, SD = 3.2). Out of the 600 children and adolescents with epilepsy 39 (6.1%) had one or more episodes during the observation period. Two factors were correlated with SE: age at the onset of epilepsy (under 5 years), and the type of epileptic syndrome (the Lennox-Gastaut syndrome, myoclonic-astatic epilepsy, symptomatic and cryptogenic partial epilepsy, progressive myoclonic epilepsy, and the Kojevnikov syndrome). Only in a fourth of the patients the cause of SP was related to a known factor, such as e.g. infection or sudden discontinuation of anti-epileptic medication. The currently used classification of SP (by Appelton and Gibbs) was found to be more useful than the one newly proposed by ILAE, because the former is better suited to the evaluation of SE in children. PMID- 12910830 TI - [Immunomorphological analysis of the vascular stroma in glioblastoma]. AB - The aim of the study was to characterize vascular stroma in glioblastoma (GB), to analyze its immunomorphological features, and to seek a relationship between patients' age and characteristics of the tumor microcirculation. Subjects in the study were 38 patients with GB, divided into two groups according to their age: group I--14 patients aged under 49 (from 32 to 49), and group II--24 patients over 50 years of age (51-78). Main types of vessels were specified: microvessels with normal morphology (capillaries, arterioles/venules), telangiectases, haemangioma--like forms, and microvascular proliferation types--simple and glomeruloid. An immunohistochemical analysis of the vascular stroma was performed with monoclonal antibodies against vWf and CD31, ASM, and Collagen IV (DAKO) to visualize the vessel wall components. The immunomorphological analysis was carried out for particular types of vessels separately. The vWf and ASM expression ratios--percentage of immunopositive area on 10 representative tumor fields under 100 x HPF--were assessed using a computer image analyzer. The incidence of some vessel types was dependent on the patients' age: in group I vascular--rich areas were more numerous, while in group II glomeruloid proliferation was more frequent--20/24-83.3% vs 8/14-57.1%. Immunoreactivity of the examined antigens was not related to the patients' age. The endothelial markers positivity was found within cells directly around the vascular lumen, and the reaction with vWf was more pronounced than CD31 reactivity. The most intense reactions were noted in proliferative changes of both types and in capillaries. ASM reactivity was found in abluminal cells within all types of vessels, most abundant in glomeruloid proliferation. ASM positivity was present also in some capillaries. Collagen IV staining revealed various abnormalities in the vascular basement membrane structure. No significant intergroup differences were found in the vWf and ASM expression ratios. However, in both groups the ASM expression ratio was significantly higher than that of vWf expression (p < 0.001). CONCLUSION: Expression of the vascular wall antigens under study depends on the blood vessel type. The number of cells with smooth muscle phenotype is significantly higher than that of endothelial cells, especially within proliferative vascular changes. Microvascular changes intensity is higher in the older patient group. PMID- 12910831 TI - [Serum and cerebrospinal fluid concentration of inflammatory proteins MIP-1-alpha and MIP-1-beta and of interleukin 8 in the course of borreliosis]. AB - Chemokines constitute a group of cytokines with a strong chemotactic action, playing an important role in the pathogenesis of inflammatory responses, including infectious meningitis. The results of in vitro experiments suggest synthesis of chemokines during Borrelia burgdorferi infection. The aim of this study was to investigate serum and cerebrospinal fluid (CSF) concentrations of the following chemokines: interleukin-8 (Il-8) and macrophage inflammatory protein 1 alpha and 1 beta (MIP-1 alpha and MIP-1 beta) in patients with neuroborreliosis. The study group consisted of 20 patients admitted to Neuroinfections and Infectious Diseases Department of the Medical University in Bialystok. The control group consisted of 12 healthy persons from whom blood samples were obtained, and 10 patients without meningitis, from whom CSF samples were taken for diagnostic purposes. Chemokine concentrations were measured with ELISA kits before treatment (baseline) and after 2 weeks of antibiotic therapy (post-treatment). Mean serum concentrations of chemokine were elevated in neuroborreliosis patients at baseline (Il-8--mean +/- SD = 668.25 +/- 661.51 pg/ml, MIP-1 alpha--124.90 +/- 89.37 pg/ml, MIP-1 beta--233.40 +/- 298.40 pg/ml) as compared to these in the control group (Il-8-23.72 +/- 7.68 pg/ml, MIP-1 alpha -36.81 +/- 4.74 pg/ml, MIP-1 beta--70.41 +/- 16.41 pg/ml). Post-treatment mean concentrations of Il-8 (197.70 +/- 285.56 pg/ml) and MIP-1 beta (102.70 +/- 42.56 pg/ml) remained significantly elevated, while the mean concentration of MIP-1 alpha (53.65 +/- 38.50 pg/ml) was insignificantly higher than that in the control group. The Il-8 mean concentration was the most elevated comparing to the controls and has decreased most significantly during the treatment. CSF concentrations of chemokines were significantly elevated both at baseline (Il-8- 754.95 +/- 535.83 pg/ml, MIP-1 alpha--24.35 +/- 4.88 pg/ml, MIP-1 beta--27.6 +/- 8.38 pg/ml) and post-treatment (Il-8--98.20 +/- 74.74 pg/ml, MIP-1 alpha--18.60 +/- 2.87 pg/ml, MIP-1 beta--16.90 +/- 4.38 pg/ml) in comparison with the controls (Il-8--10.43 +/- 2.70 pg/ml, MIP-1 alpha--8.17 +/- 1.54 pg/ml, MIP-1 beta--7.27 +/- 1.58 pg/ml). MIP-1 alpha and MIP-1 beta CSF concentrations were significantly lower than their concentrations in serum. The Il-8 CSF concentration did not differ significantly from its serum concentration. However, in some patients Il-8 CSF concentration was much higher than that in the serum, which suggests its significant synthesis within the cns and its role in the pathogenesis of B. burgdorferi meningitis. Chemokine CSF concentrations were not correlated with cytosis and CSF protein concentration. The results indicate the induction of Il 8, MIP-1 alpha and MIP-1 beta synthesis in the course of neuroborreliosis and a decrease of their concentrations during 2 weeks of treatment, however, without reaching the normal values. PMID- 12910832 TI - [Awake craniotomy and brain mapping for eloquent cortex in patients with supratentorial tumors: a preliminary report]. AB - Malignant brain tumours are incurable at present. Since none of the hitherto used treatment methods allows to significantly extend these patients' survival time, the basic aim is to improve their quality of life. Intraoperative brain mapping seems to be an approach enabling to minimize the risk of irreversible damages to functionally important structures of the brain. In the Department of Neurosurgery of the Medical University of Lodz awake craniotomy with stimulation of eloquent cortex was attempted from May 1999 to July 2000 in 13 patients aged 16 to 77 years. In two patients the attempt of intraoperative awakening was unsuccessful. In 8 out of the 11 awakened patients intraoperative brain mapping had a significant effect on the course of surgery (i.e. on the resection magnitude and "safe corridor"). Out of six patients with tumors situated in the neighbourhood of motor cortex--one developed a severe and permanent paresis of the upper limb. On the grounds of the literature and the authors' own experience an algorithm of awake craniotomy and intraoperative brain mapping was worked out. PMID- 12910833 TI - [Neuro-endoscopic techniques in the treatment of complex hydrocephalus in children]. AB - The aim of the paper was to evaluate effectiveness of neuroendoscopic procedures in comparison to complex shunt systems implantation in the treatment of complex compartmentalized hydrocephalus in children. Neuroendoscopic techniques were applied in 47 patients (23 boys, 24 girls aged from 25 days to 18 years, mean age 3 years SD = 4.9 years). The shunt implantation comparison group consisted of 80 patients (47 boys and 33 girls aged from 1 day to 16 years, mean age 0.8 year, SD = 2 years). Every endoscopic procedure was planned individually, in accordance with the patient's type of complex hydrocephalus and level of deformity of his/her ventricular system. In cases of multiloculated hydrocephalus septostomy was generally performed to restore communication between separated parts of the ventricular system. In uniloculated hydrocephalus resulting from the foramen of Monro obliteration, septostomy of pellucid septi was performed to connect the isolated lateral ventricles. In cases of isolated ventricle III the foramen of Monro patency was restored to connect the ventricle with the whole ventricular system. Neuroendoscopic techniques allowed to significantly reduce the number of necessary surgical procedures. Complex hydrocephalus patients treated with the traditional shunt implantation required on the average 7 operations during the whole therapy, as compared to about 2 in those treated by means of neuroendoscopic techniques. An analysis of the number of necessary surgical interventions per year of clinical observation also indicated superiority of neuroendoscopy techniques over shunt implantation (1 vs. 4 operations per year, respectively). Neuroendoscopy allowed to simplify shunt systems in 33 children, i.e. 70.2% of those treated with neuroendoscopic techniques, while in the group treated with traditional methods of shunt implantation only 16 children (16.3%) had a simple shunt system (a shunt with one intraventricular drain). The outcome assessed according to the Glasgow Outcome Scale (GOS 1, 2) in children treated only by shunt implantation was significantly inferior to that in the neuroendoscopy group, both in terms of mortality rate (22.5 and 4.3%, respectively) and postoperative complications present in 42 (52.5%) of shunt implantation cases and 9 (19.1%) of endoscopically treated patients. CONCLUSIONS: Neuroendoscopic techniques allowed to reduce the number of necessary operative procedures, to simplify shunt systems, to improve clinical outcome, and to reduce the risk of complications in the early postoperative period. PMID- 12910834 TI - [Angio-MR assessment of vasomotor reactivity impairment as a long-term outcome of SAH following intracerebral aneurysm rupture]. AB - Studies on long-term outcome of subarachnoid hemorrhage (SAH) have been carried out for many years using various neuroimaging techniques, such as e.g.: SPECT, PET, TCD and XeCT. In our study angio-MRI supplemented with the acetazolamide test was used to assess cerebrovascular reserve impairment in 30 patients within 6 months since clipping an intracranial aneurysm. Severity of the SAH course was evaluated using the WFNS (World Federation of Neurosurgical Societies) scale [3]. The patients' clinical status was assessed at follow-up by means of the Glasgow Outcome Scale (GOS) [8]. Cereberovascular reserve evaluated at the follow-up in hypercapneal conditions was found to be insufficient. The degree of vessel reactivity dysfunction as a long-term outcome of SAH turned out to depend on massiveness of hemorrhage from the ruptured aneurysm. PMID- 12910835 TI - [A study of lyophylized human brain tumor tissue using MAS 1H-MRS spectroscopy]. AB - AIMS: Proton magnetic spectroscopy of brain tumor extracts, and more recently magic angle spinning proton magnetic resonance spectroscopy (MAS 1H-MRS) of intact cooled tissue specimens have been used for tumor classification. The aim of this study was to obtain MAS 1H-MRS spectra of lyophilised and pulverised specimens of various brain tumors and to assess their utility for tumor type recognition. MATERIAL AND METHODS: Tissue samples of intracranial tumors were obtained at surgery and frozen immediately in liquid nitrogen. A part of each sample was subjected to a routine histopathological examination, while the rest was lyophilised, pulverised, and subjected to high-speed magic angle spinning MRS using Bruker DSX-400 WB AVANCE spectrometer and rotation speed up to 33 kHz. RESULTS: MAS 1H-MRS spectra of 39 samples (including 16 glioblastomas or gliosarcomas, 10 meningiomas and 5 craniopharyngiomas) were obtained. Invariably the spectra were dominated by broad lipid resonances, reflecting "NMR visible" protons in methyl,methylene and methine groups of fatty acid chains. Resolution of the spectra was markedly higher for glioblastomas/gliosarcomas and craniopharyngiomas than for meningiomas. CONCLUSIONS: Lyophylization of brain tumor specimens eliminates signals of watersoluble metabolites and results in a loss of discriminative power of the 1H-MRS analysis. Resolution of the spectra is increased for tumors that accumulate lipids extracellularly, but is not related to tumor histological grade. PMID- 12910836 TI - [Comparison of effectiveness of percutaneous embolization and microsurgery in the treatment of 60 patients with MCA berry aneurysms]. AB - Surgical clipping has been considered for years a "golden standard" in the treatment of middle cerebral artery (MCA) aneurysms. The recent development of materials and endovascular techniques has permitted the use of embolization for the treatment of intracranial aneurysms regardless of their locations and size. In 30 patients with 32 MCA aneurysms (group I) the endovascular approach with GDC 10 coils was used, while in other 30 patients with 31 aneurysms (group II) surgical clipping was performed. On completion of the diagnostic process the patients were randomly assigned either to the endovascular or microsurgical treatment. Group I: 21 (70%) embolization procedures were carried out, while in 9 (30%) cases with 10 aneurysms embolisation was impossible or regarded as too risky due to various causes. The total occlusion of aneurysm SAC was achieved in 17 (77%) aneurysms, subtotal in 5 (23%). The morbidity rate was 8%, mortality-0%. Group II: Microsurgical clipping was performed in 27 (90%) cases, 3 (10%) patients were disqualified from treatment due to their poor clinical status. An excellent technical result was achieved in 24 (86%) patients, in 4 (14%) cases the neck remnant was observed in control angiography. The morbidity rate was 14%, mortality-0%. PMID- 12910837 TI - [Facio-scapula-humeral muscular dystrophy: clinical picture and molecular genetics]. AB - Facioscapulohumeral muscular dystrophy (FSHD) is a disorder of muscle with a progressive, often asymmetric wasting and weakness of facial, shoulder girdle and lower limbs muscles. No FSHD gene has been identified so far. The FSHD locus is known to be 4q35. The paper presents the clinical picture of FSHD including atypical cases, as well as its inter- and intrafamilial clinical variability. Molecular pathology and diagnostics of the condition are discussed, with particular attention paid to DNA analysis in FSHD. PMID- 12910838 TI - [Neurophysiological testing in myasthenia syndromes]. AB - Myasthenic syndromes are a heterogeneous group of congenital or acquired disorders of neuromuscular junction. Despite major advance in genetics and molecular biology of disorders of neuromuscular junction, clinical diagnosis and choice of treatment largely depends on results of neurophysiological tests. Different protocols of repetitive nerve stimulation and single fibre EMG are indispensable in confirming neuromuscular junction defect, they can also give additional information on the level of abnormality and differentiate myasthenia gravis from Lambert-Eaton syndrome (pre- or postsynaptic defect). Characteristic features of repetitive nerve stimulation test e.g. repetitive response allow diagnosing congenital myasthenic syndromes such as slow channel syndrome or acetylcholine deficiency. Patophysiological basis of neurophysiological tests of neuromuscular transmission is presented. Different neurophysiological findings in cases of Lambert-Eaton myasthenic syndrome and congenital myasthenic syndromes are presented. PMID- 12910839 TI - [Fronto-temporal dementia and parkinsonism linked to chromosome 17 (FTDP-17): clinical criteria]. AB - Frontotemporal dementia and Parkinsonism linked to chromosome 17 (FTDP-17) is a congenital degenerative disease of the central nervous system. Molecular genetics studies have indicated the presence of 26 various mutations of the tau protein gene. At present 61 families with this syndrome are known. The clinical criteria proposed in this paper were developed on the grounds of the author's own research as well as detailed review of the literature. The criteria include;--clinical symptoms: personality and behavior disorders, Parkinsonism rather resistant to treatment with Levodopa less frequently also speech disorders and epilepsy,--an early onset and rapid progression of the disease,--positive family history,- heterogeneity of clinical symptoms both among patients within the same family and among families with the same mutation,--heterogeneity of the clinical picture depending on the type of mutation. Although FTDP-17 is and extremely rare clinical syndrome, it is noted all over the world. In Poland nobody has been diagnosed with this condition yet. The aim of this paper is to acquaint the readers of Neurologia i Neurochirurgia Polska with the FTDP-17. The syndrome has considerably contributed to our understanding of pathogenesis of many sporadic degenerative diseases of the brain, including such frequent conditions as Alzheimer's disease, Pick's disease the the Steel-Richardson-Olszewski syndrome, corticobasal degeneration, and other ones. Undoubtedly further research into the FTDP-17 will contribute to the development of a successful treatment for these devastating degenerative diseases of the c.n.s. PMID- 12910840 TI - [The importance of transcranial Doppler (TDC) in the assessment of cerebrovascular hemodynamics of the acute phase of ischemic stroke]. AB - Most recent studies on transcranial Doppler (TCD) in the acute phase of ischemic stroke are reviewed in this paper. TCD is a highly sensitive and specific method of quick, bedside assessment of cerebrovascular circulation hemodynamics in the acute phase of ischemic stroke. The following issues are discussed in the paper: a new classification of ultrasound pathological changes associated with intracranial arteries occlusion or stenosis, the frequency of spontaneous re canalisation by TCD, a comparison of TCD results with findings obtained by means of other vascular imaging techniques (e.g. DSA, CTA and MRA), relationship between TCD baseline assessment of primary blood flow changes and thrombolysis efficacy (the so-called TIBI classification, Thrombolysis in Brain Ischemia), and prognostic significance of TCD in the acute ischemic stroke. Moreover, first clinical reports on an additive effect of TCD on thrombolytic therapy are presented. The main limitations of TCD include the lack of sufficient bone window in 5-15% of patients and the lack of sufficiently trained staff capable of performing the examination a 24 h duty, since results of the examination to a large degree depend on the examiner's skills. PMID- 12910841 TI - [The currently accepted pathophysiological model underlying surgical management of Parkinson disease]. AB - Among all the extrapiramidal movement disorders Parkinson's disease (PD) is the one most often submitted to neurosurgical treatment. Technical advances in neurosurgery, neuroimaging and neurophysiology, as well as shortcomings of chronic Levodopa medication (i.e. on/off fluctuations, violent dyskinesia and painful dystonia) have greatly contributed to the renewed interest in the surgical treatment of PD. The attainment of a better understanding of the basal ganglia function and of PD pathophysiology has also encouraged centers to treat Parkinson's disease in recent years. This article presents the current model of PD and the rationale for using GPi, thalamus and STN as target sites in stereotactic surgery. PMID- 12910842 TI - [Stereotactic surgery in Parkinson disease: patient selection criteria in the light of existing research]. AB - The renewal of interest in the neurosurgical treatment of Parkinson's disease (PD) also in Poland results from of our improved understanding of functional anatomy of basal ganglia, developments in neurophysiological and neuroimaging techniques, as well as from advances in stereotactic surgery techniques. In view of the growing number of PD patients referred to surgical treatment and the wide variety of interventions offered, the development of clear of PD patient's selection to surgery seems necessary. Various surgical options and possible targets provide different functional benefits, but the almost 10 year's experience makes us aware also of the limitations and possible complications involved. Algorithms worked out by researchers from the most experienced centers include the following selection criteria as a minimal standard of the PD patient's evaluation before surgery: a reliable diagnosis of Parkinson's disease, at least 5 years of PD duration since the onset of symptoms, good responsiveness to L-dopa or apomorphine, exclusion of severe depression and dementia, neuroimaging (MRI) performed before the surgery, and optimal (but ineffective) attempts at available pharmacological therapy prior to the surgery. PMID- 12910843 TI - [A case or Leber hereditary optic neuropathy (LHON): differential diagnosis with post inflammatory atrophy of nerve II using the mtDNA analysis]. AB - The Leber hereditary optic neuropathy (LHON) is a disease due to a mtDNA mutation. The disorder results from enzymatic perturbations in the mitochondrial respiratory chain. Clinically the LHON may present as a progressive axonal atrophy of the optic nerves with or without other neurological symptoms. The process of reaching the diagnosis of the LHON by means of the molecular analysis of mtDNA is discussed. PMID- 12910844 TI - Huntington's disease, case report. AB - Huntington's disease (HD) is a chronic neurodegenerative disorder, characterized by the following triad of clinical hallmarks: chorea, cognitive impairment and behavior disorders [8]. In 1993 the gene responsible for HD, whose mutation results in HD, was identified and mapped on the chromosome 4p16.3 [6]. The mutation is a characteristic expansion of a CAG nucleotide triplet. In this paper we present a 36-years-old female patient with HD who was submitted to a complete diagnostic procedure including genetic testing. Her pedigree was reconstructed using available medical documentation and tracing other members of her family. PMID- 12910845 TI - [Guillain-Barre syndrome following Herpes zoster infection]. AB - A male patient with a bi-phasic Guillain-Barre syndrome is presented in the paper. Initially a paralysis of his left lower extremity was observed, with hypotonia and removal of deep reflexes, followed by a flaccid paralysis of the remaining extremities. This syndrome had been preceded by a Herpes zoster infection producing symptoms in the left shank and foot. The paresis of his left lower extremity had been increasing during the week before his admission, while the paralysis of the other extremities occurred not earlier than by the end of the second week of his hospitalization. The diagnosis of the Guillain-Barre syndrome was based on the clinical course of the disease, and results of EMG examination and of general laboratory tests of his cerebrospinal fluid. PMID- 12910846 TI - [Primary leptomeningeal sarcomatosis. Case report]. AB - Primary intracranial sarcomas are very rare and aggressive tumors. A case of a 40 year-old woman with primary leptomeningeal sarcomatosis is presented. Initially her clinical symptoms resembled those of a pseudo tumor cerebri. Neuroimaging did not allow establishing a diagnosis of leptomeninges neoplastic infiltration. The patient died 8 months since the onset of symptoms. A neuropathological examination revealed diffuse thickening of leptomeninges. Microscopically a widespread sarcomatous infiltration of the subarachnoid space was shown, as well as a neoplastic infiltration of the choroid plexus and of some perivascular spaces in the cerebral cortex. On the grounds of a review of the literature diagnostic problems, differential diagnosis and clinical course of primary leptomeningeal sarcomatosis are discussed. PMID- 12910848 TI - [Obituary to Professor Wanda Horyd]. PMID- 12910847 TI - [Trigeminal neuralgia due to lengthy AVM treatment. A case report]. AB - The authors presented a case of iatrogenic trigeminal neuralgia in a 46-year-old man with a dural arteriovenous malformation (AVM) previously treated by means of intravascular embolization. At first, microvascular decompression (MVD) in the posterior fossa was performed, but unfortunately the pain persisted. Subsequently, percutaneous stereotactic radiofrequency rhizotomy was carried out. The procedure was uneventful and brought about an immediate pain relief. PMID- 12910849 TI - [Report on the 1st International Frankfurt advanced skull microanatomy and hand on dissection workshop, Frankfurt/Maine, October 17-19th]. PMID- 12910850 TI - [Report on the Fifth Congress of the European association of neuro-oncology, Florence, Italy, September 7-10, 2002]. PMID- 12910851 TI - [Report on the 18th Congress of Polish Neurological Society]. PMID- 12910852 TI - [Prevalence of depression in neurological outpatients. DEPEND study]. AB - For the assessing the incidence of mood disturbances among the neurological out patients 3287 of them were examined by 111 neurologists during their routine practice. Early diagnosis, the type of mood disturbances and the depth of depression were estimated by the use of Beck's Depression Inventory, the questionnaire based on The Mini-International Neuropsychiatric Interview and Hamilton Depression Rating Scale, as well. Around half of the patients (50.47%) were suspected on depression, as an early diagnosis. In suspected and diagnosed depressive patients the symptoms as anxiety, low activity precordial pain, headaches, dry mouth, constipation, sleep and appetite troubles were significantly (p < 0.01) more frequent than in euthymic subjects. Among all studied patients the episode of depression were found as a final diagnose in 17.2%, recurrent depressive disorders--in 17.6% and dysthymia--in 2.8% of subjects. In finally diagnosed depressive patients the chronic neurological problems were significantly (p = 0.013) more frequent, as the cause of the visit, than in the euthymic ones. The low mood was equally frequent among the patients with Parkinson's disease, multiple sclerosis and cerebrovascular disorders, as well. PMID- 12910853 TI - [Prevention of stroke and vascular dementia]. AB - Prevention of stroke and dementia seems to be a great and rising challenge nowadays. Increasing problem of cardiovascular diseases considers, apart from individuals' and state expenses, also important social effort. The aspect of prevention seems to be the most optimistic accent in that area. Based on the results of multiple randomized trials we can influence in both pharmacological and non-pharmacological way the major risk factors of stroke. Effective antihypertensive treatment, cholesterol lowering therapy, smoking cessation and antithrombotic treatment in subjects with atrial fibrillation result in noticeable reduction of stroke events in both primary and secondary prevention. The number of researches which aim the dementia prevention is still sparse, which makes the results of the recently presented PROGRESS trial even more interesting and valuable. It showed that it was possible to prevent dementia and cognitive impairment with therapy based on the long acting ACE inhibitor. An increasing number of data supporting the beneficial role of statins in the stroke and dementia prevention it is noteworthy. The multifactorial intervention both in prevention of stroke and dementia seems to be the most effective. PMID- 12910854 TI - [Depression and dementia]. AB - Specific symptoms of depression in aged subjects are presented in the paper. Cognitive dysfunctions in elder depressive patients (so called pseudodementia) as well as comorbidity of dementia with depression are special diagnostic problems. Both etiopathogenetic and clinical issues are discussed. PMID- 12910855 TI - [49-year-old patient with staphylococcal endocarditis and thyrotoxic crisis]. AB - We report the case of a 49-year-old woman who presented with typical features of thyroid crisis. Besides the confirmation of severe thyreotoxicosis a native valve endocarditis due to Staphylococcus aureus was diagnosed. Diagnostic criteria and therapies of these life-threatening disorders are discussed and a possible association in the present case is evaluated. PMID- 12910856 TI - [Unilateral pulmonary edema in a dialysis patient with massive fluid overload and mitral valve insufficiency]. AB - A pulmonary edema is not a rare complication in patients with chronic renal failure. Arterial hypertension and fluid overload play a pathogenetically important role. The diagnosis of an unilateral pulmonary edema is often difficult. The following arguments favour the diagnosis: rapid clinical amelioration and disappearance of the alveolar opacities after diuretic treatment alone within 24-72 hours. We describe the case of a 37-year-old patient on dialysis treatment who showed a pulmonary edema of the right lung, predominantly in the upper lobe. This edema was due to fluid overload and mitral insufficiency. After diuretic treatment alone the patient rapidly recovered and the lung infiltrates on the chest X-ray disappeared within a few days. PMID- 12910857 TI - [Cholestatic hepatitis. 27-year-old tool engineer]. PMID- 12910858 TI - [Invasive and noninvasive techniques for analysis of cardiovascular effects of sleep apnea]. AB - There is increasing epidemiological and experimental evidence that Sleep Disordered Breathing (SDB) is associated with cardiovascular disease such as hypertension, ischaemic heart disease, heart failure and stroke. Due to the high prevalence of SDB in the general population (5% to 10%) there is increasing demand for cost-effective and reliable diagnostic tools for the assessment of cardiovascular function during sleep in patients with SDB. The first part of this review focuses on our present knowledge about the association between SDB and cardiovascular disease. In the second part various methods for the assessment of cardiac function, blood pressure, sympathetic activity, as well as vascular and cerebrovascular function in patients with SDB are discussed. Current developments such as ECG analysis for SDB screening or arterial tonometry are introduced. Further improvements in the diagnostic tools for the investigation of cardiovascular function in patients with SDB may to advantage be coupled with epidemiological studies. This approach may demonstrate the predictive value or superiority of a specific diagnostic parameter in the diagnosis of SDB and its cardiovascular consequences. PMID- 12910859 TI - Determination of lesion size by ultrasound during radiofrequency catheter ablation. AB - The catheter tip temperature that is used to control the radiofrequency generator output poorly correlates to lesion size. We, therefore, evaluated lesions created in vitro using a B-mode ultrasound imaging device as a potential means to assess lesion generation during RF applications non-invasively. Porcine ventricular tissue was immersed in saline solution at 37 degrees C. The catheter was fixed in a holder and positioned in a parallel orientation to the tissue with an array transducer (7.5 MHz) app. 3 cm above the tissue. Lesions were produced either in a temperature controlled mode with a 4-mm tip catheter with different target temperatures (50, 60, 70 and 80 degrees C, 80 W maximum output) or in a power controlled mode (25, 50 and 75 W, 20 ml/min irrigation flow) using an irrigated tip catheter. Different contact forces (0.5 N, 1.0 N) were tested, and RF was delivered for 60 s. A total of 138 lesions was produced. Out of these, 128 could be identified on the ultrasound image. The lesion depth and volume was on average 4.1 +/- 1.6 mm and 52 +/- 53 mm3 as determined by ultrasound and 3.9 +/- 1.7 mm and 52 +/- 55 mm3 as measured thereafter, respectively. A linear correlation between the lesion size determined by ultrasound and that measured thereafter was demonstrated with a correlation coefficient of r = 0.87 for lesion depth and r = 0.93 for lesion volume. We conclude that lesions can be assessed by B-mode ultrasound imaging. PMID- 12910860 TI - [A novel device for active shoulder rehabilitation with glenohumeral traction or compression]. AB - The equipment available for rehabilitation of patients with joint conditions still lacks a device that, in common with usual physiotherapeutic measures (manipulation, proprioceptive neuromuscular facilitation, Bobath therapy), is based on traction and/or compression or oscillating forms. The novel "Shoulder Rotator" is such a simple device for the variable treatment of shoulder disorders in all three spatial dimensions. After individual instruction, the patient performs rotating glenohumeral exercises with simultaneous glenohumeral joint traction, compression, or oscillations against static or dynamic resistance. The "Shoulder Rotator" is similar to the established continuous passive motion device for the shoulder and is mounted on a mobile chair. The electronic device control is computerised and permits a wide variety of treatment options, individual regimes, and documentation. The oscillating, compression/distraction mode of operation provides an analgesic effect. The isolated compression mode along the rotatory axis enables conditioning of joint proprioception. The distraction mode along the rotatory axis facilitates muscular relaxation. Resistive rotatory exercises strengthen the rotatory glenohumeral muscles and help centre the humeral head in the glenoid. In addition to the advantage of adding documentation of treatment progress as required for evidence-based medicine, this novel device may bring about a cost-effective, genuine quality improvement in shoulder rehabilitation. PMID- 12910861 TI - [Mechanical versus drug prevention of thrombosis after total hip endoprosthesis implantation. A randomized, controlled clinical study]. AB - Pharmacological prophylaxis is routinely applied after total hip replacement. Although it effectively reduces deep-vein thrombosis, side effects (bleeding, haematoma, swelling, thrombocytopenia) are not infrequent. Since in Germany use of foot pumps as only means of prophylaxis is unpopular, we investigated their efficacy and safety in a randomized study. 106 patients used either low molecular weight heparin (Fraxiparin, Sanofi-Synthelabo, Germany) or the foot-pump (A-V Impulse System, Orthofix, Muhltal, Germany), and were monitored for deep-vein thrombosis using serial duplex sonography on postoperative days 4, 12 and 45. Clinical observations included daily measurements of thigh circumference, recording of postoperative drainage amounts, and monitoring of wound healing. None of the 50 patients treated with the foot-pump developed deep-vein thrombosis, while 4 of the 50 patients (8 per cent) on pharmacological prophylaxis did so. Six patients stopped using the foot-pump during the study. One patient developed heparin-induced thrombocytopenia. Patients on mechanical prophylaxis had smaller amounts of drainage (mean 247 ml vs. 272 ml, p = 0.485) and significantly less swelling of the thigh (10 mm compared with 15 mm, p or = < 0.001), The good results in terms of prevention of thromboembolic complications and soft tissue swelling favour the general use of foot pumps as mechanical prophylaxis. PMID- 12910862 TI - [Biomechanical design of load simulation in multiple spinal segments]. AB - Multisegmental biomechanical studies on the lumbar spine are steadily increasing in importance. Only in this way can we acquire knowledge about the physiological behaviour of the entire lumbar spine. Furthermore, these studies allow us to analyse in vitro the biomechanics of manipulated lumbar spines after various surgical operations on the spine. A load simulator was developed to investigate multisegmental lumbar spine mobility, and its function was investigated in an initial study on 19 fresh--frozen specimens of human lumbar spine. After x-ray examination and determination of the bone mineral density, the specimens were loaded up to 10 Nm in the automatic electromechanical loading system under flexion/extension, lateral bending and axial rotation. An ultrasound-based motion analysis system was used to measure the displacements of the vertebrae involved. PMID- 12910863 TI - [Histologic and histomorphometric follow-up observations of osseointegration of corundum-blasted BMP-3 coated titanium test implants (Ti6Al4V) at orthotopic site in the giant rabbit]. AB - AIM: To establish whether the additional coating of titanium implants with Bone Morphogenetic Protein-3 (BMP-3) might enhance osseous integration. METHOD: Each of 15 cylindrical titanium test implants (Ti-6AI-4V) was coated using 230 micrograms porcine BMP-3. A further 15 implants with identical (corundium blasted) surface served as negative controls. An uncoated and a BMP-3-coated test object were implanted into the femurs of 15 adult giant rabbits. New formation of bone around the implants was examined microscopically and histomorphometrically on postoperative days 14, 35 and 56. RESULTS: Coated implants revealed superior osseointegration with statistical evaluation using the t-test for matched samples showing a significantly higher volume of new bone 5 weeks after surgery. Microscopic examination revealed osseointegration with no connective tissue membrane around the surface of the implants. CONCLUSIONS: Our results indicate that composite metal implants are suitable carriers for BMP-3 and that improved fixation of the implants can be achieved. PMID- 12910864 TI - [Thermal injury and wound healing of the endometrium subsequent to minimally invasive transendoscopic use of Nd:YAG-laser-and electrosurgery in horses]. AB - Transendoscopic thermal preparation techniques like laser- or electrosurgery have proved to be most reliable for minimal invasive intrauterine surgery during operative hysteroscopy in mares. To determine the effect of Nd:YAG laser surgery on the endometrium and the complete uterine wall compared with electrosurgery, standard lesions were obtained by transendoscopic monopolar electrosurgery (loop electrode, cutting blade) and Nd:YAG laser surgery (contact, bare fibre, 25 Watt, exposure time four seconds, non contact, 80 Watt, exposure time four seconds) in five healthy mares on days d 0, 7, 14, 18 and 21. The gross effects were controlled endoscopically before the consecutive lesions were set. Following the last diathermic endometrial irritation, hysterectomy was performed under general anaesthesia. Recovery of the mares was uneventful. Specimens from the lesions were collected after dis-section of the uterus, fixed in formalin, routinely embedded and stained with H. E. for histological evaluation. The different energy forms caused tissue alterations of comparable quality but differing in degree. Most intensive peripheral hyperemia and tissue edema occurred on postoperative days three to seven. Non contact laser irradiation initiated maximal amount of fibrinous exudate. Lesions created with a loop electrode were superficial only and did only affect the endometrium. Preparation with an electrosurgical cutting blade or a laser bare fibre resulted in craterlike lesions lined by carbonisation and did involve deeper layers of the myometrium. The lesions caused by non contact laser irradiation affected all layers of the uterine wall. Three weeks postoperatively, reepithelization of the luminal epithelium was completed. The depth of thermal injury and coagulative necrosis, inflammatory reactions and scar formation was greater in lesions created by laser application compared with those set with a loop electrode at power densities comparable to those usually achieved at hysteroscopic surgery. The results suggest that an electrosurgical loop designed for operative hysteroscopy may be a reliable tool for diathermic resection of pathological endometrial structures like uterine cysts characterized by minimal thermal injuries and a short period of reconvalescence. Non contact laser irradiation may result in deep thermal injuries with the risk of delayed uterine perforation but may be profitable for treatment of partly intramural structures like an intramural leiomyoma. PMID- 12910865 TI - [Pulmonary wedge pressure and heart frequency measurements during standardized treadmill exercise for extension of left atrial function diagnosis in warmblood horses]. AB - In 12 healthy warmblood horses (six trained and six untrained) the pulmonary wedge pressure and heart frequency was measured at rest and during a standardised exercise test on a treadmill. The mean pulmonary wedge pressure at rest was 14.53 +/- 2.36 mmHg. There was no significant difference in pulmonary wedge pressure either at rest or during exercise between trained and untrained horses. During walking (1.8 m/s) the mean pulmonary wedge pressure was 19.62 +/- 4.03 mmHg, during trotting (4 und 5 m/s) it was between 22.38 +/- 3.92 mmHg and 25.28 +/- 3.7 mmHg. During canter (6 m/s) and gallop (8 m/s) the mean pulmonary wedge pressure increased to a level of 25.54 +/- 4.3 mmHg and 31.86 +/- 4.29 mmHg. There was a significant increase in pulmonary wedge pressure with each incremental step of the standardised treadmill test. Concerning mean heart frequency a highly significant increase could be observed at the beginning and at the end (treadmill speed of 7 and 8 m/s) of the standardised exercise test. At higher intensity of the exercise test (7 m/s and 8 m/s) untrained horses showed a significantly increased heart rate compared to trained horses. Neither at rest nor during the different exercise levels a significant correlation factor greater 0.5 between heart frequency and pulmonary wedge pressure could be observed. The increase of heart frequency and pulmonary wedge pressure during exercise showed no correlation. Between left atrial size and pulmonary wedge pressure a statistical weak correlation could be observed up to a treadmill velocity of 6 and 7 m/s. PMID- 12910866 TI - [Disinfection of caliciviruses at 20 and 10 degrees C]. AB - Five disinfectants, Venno FF super, Venno Vet 1 super, Venno Oxygen, M&Enno Veterinar B neu und Neopredisan 135-1, were tested to evaluate their efficacy against caliciviruses at 20 and 10 degrees C. As model test virus served feline calicivirus type F9 (FCV F9). All disinfectants were tested according to Guidelines of the German Veterinary Association (DVG). The investigations were performed in suspension tests and germ carrier tests. The suspension tests were carried out without and with protein load. As protein was used foetal calf serum at the concentration of 40%. Venno FF super showed less protein dependence, however a considerable temperature dependence. This matter can be corrected by increase of concentration on 2%. Venno Vet 1 super was without protein especially effective. The losses on the effectiveness through low temperature and protein load can be annulled also here by increase of concentration. Venno Oxygen was more effective in the comparison to that here named both preparations. The effects of temperature can be corrected by extension of reaction time. The most effective preparation was M&Enno Veterinar B neu. The disinfection occurred at 20 degrees C with 0.5% solution within 120 min and at 10 degrees C with 1.0% solution within 60 min. The fifth disinfectant Neopredisan was in suspension tests without protein load and carrier tests with gauze at 20 and 10 degrees C relative convincing but in germ carrier tests with poplar wood, no complete disinfection could be achieved within tested concentrations and reaction times. PMID- 12910867 TI - ["Emergency splitting" of the teat canal--a therapeutic procedure in case of concurrent occurrence of acute thelitis and severe mastitis]. AB - In case of concurrent acute thelitis and severe mastitis, an immediate surgery of the stenosis ("emergency splitting") might be justified in order to improve milkability as a prerequisite for the removal of pathological milk secretion and therapy, successful mastitis therapy. The aim of this study was to compare the milkability and the clinical cure rate of mastitis between Group A (n = 19, cows with stenosis, thelitis and mastitis) and Group B (n = 19, cows with stenosis, with or without clinical mastitis, but without acute thelitis) following surgery of the papillary duct using a cutting instrument (Danish cannula model with a double cutting edge). In most cases, milkability was restored at the end of the treatment in both groups (Group A: 89.5%; Group B: 100%). In this time period nine of 19 quarters with mastitis in Group A were clinically cured (47%). Six month after the surgery, the animal owners considered the milkability as very good in 70% and 72% of the cases in Group A and Group B, respectively. However, "emergency splitting" led to a more intensive and longer postsurgical treatment. The average duration of total treatment (day of surgery plus 6 days of obligate postsurgical treatment plus additional treatment days) was 7.4 and 6.7 days in Group A and Group B, respectively. Furthermore, the higher expenditure was due to the more costly mastitis therapy. During the six month following surgery, only one cow of Group A had to be culled due to persistent mastitis and two cows because of insufficient milkability. PMID- 12910868 TI - Crowding and winter emergency feeding as predisposing factors for kerato conjunctivitis in semi-domesticated reindeer in Norway. AB - Due to the hard environmental and climatic situation in late winter 1999, a herd of about 200 free-ranging, semi-domesticated reindeer was gathered in a paddock in northern Norway for emergency feeding. About the same number of reindeer was not corralled but supplementary fed on their winter pastures. The fodder was of relatively good quality but very dusty and fed in a very dry environment. Six weeks later, an outbreak of eye-infection was diagnosed in one third of the corralled reindeer; mild symptoms were observed in most of them, but 11 animals showed severe signs of disease. No signs of disease were found in the non corralled animals. Ten reindeer died through emaciation, the eleventh was sacrificed. Histopathological diagnosis of two severely affected eyes revealed a severe purulent kerato-conjunctivitis with bacteria and plant particles embedded in purulent exudates on the cornea and conjunctiva. In one eye from the two most affected animals Actinomyces pyogenes, coagulase-negative Staphylococci and Escherichia coli and in the other one Staphylococcus aureus and Escherichia coli were found. The bacteria encountered in this study are not considered the primary cause of disease. They seem rather to be opportunistic infectious agents of eyes that have been irritated mechanically through exposure to dusty fodder in a dry environment. The stress through unfamiliar corralling of the reindeer, that followed an insufficient fodder supply, could be considered as an additional infection supporting factor. This case-report emphasises on the importance of different factors involved in favouring outbreaks of disease in reindeer, under intensified husbandry conditions. Even though crowding and emergency feeding may be, at certain circumstances, the only means of survival for reindeer, a negative impact of implied crowding diseases on their productivity, must be considered, as well. PMID- 12910869 TI - Considerations for gastrointestinal cannulation (rumen, duodenum and ileum) in sheep with a ruminal, a simple t-type and a modified t-type cannula. AB - The aim of this study was to investigate the feasibility of simultaneous application of three cannulations and to test the effectiveness of a modified T type cannula compared to simple T-type cannula. A ruminal, a simple T-type and a modified T-type duodenal and an ilial cannula were used in sheep. No complication was encountered in ruminal cannulation. Although no grafts were used, leakage of digesta was not seen for at least five months in T-type cannula and seven months in modified T-type cannula after surgery. Fibrous tissue grew between the serosa of the intestine and the peritoneum. No erosion of the tissue between intestine and body wall was observed after slaughter. Especially ilial digesta was easily obtained from modified T-type cannula. PMID- 12910870 TI - [Animal nutrition for veterinarians--actual cases: tulip bulbs with leaves (Tulipa gesneriana)--an unusual and high risk plant for ruminant feeding]. AB - 14 cattle (mainly younger ones) of a total of 50 extensively kept Galloways died within 6 weeks in late winter 2001/02. According to the owner's report, grass growth had been rather poor; therefore, the herd was fed additionally hay as well as large amounts of tulip onions. In the microbiological examination a highly reduced hygienic quality of the roughage could be detected. In the rumen contents of two dissected young cattle parts of tulip onions were found. According to pertinent literature, tulip onions (in particular their external layers) contain variant-specific amounts of anti-nutritive substances; main active agents are tulipin (a glycoprotein), tuliposid A and B, and lectins. They may cause intensive mucosal irritation, accompanied by reduced feed digestion and body weight gains, drooling, vomiting and diarrhea. This case report underlines risks caused by feeding of plants originally not destined as forage, if their active ingredients and effects are unknown or remain unconsidered. PMID- 12910872 TI - [Persistence of circulation of Corynebacterium diphtheriae non-toxigenic strains and low prevalence of carriers in a sample of children vaccinated against diphtheria]. AB - This study was carried out with the aim to investigate the presence of carriers of diphtheria bacillus in a sample of 1970 healthy children, 6-14 years aged, residing in eight Italian towns. Three non-toxigenic strains of Corynebacterium diphtheriae biotype mitis were isolated from as many healthy children. Molecular characterization by ribotyping showed close genetic relation of two of the wild strains with the C7(b) reference strain whereas one of the wild strains showed close genetic relation with two collection strains isolated in the same geographic area (Emilia-Romagna) from diphtheria patients in the seventy years. This supports the hypothesis of the persistence of some non toxigenic C. diphtheriae strains derived from ancient endemic strains under the selective pressure of mass immunization against diphtheria. The persistence of carriers of diphtheria bacilli, although non toxigenic, suggests that high levels of immunity must be maintained, not only in children, but also in adults by booster vaccination. PMID- 12910871 TI - [Prevalence of monoclonal gammopathies: an experience in Western Sicily]. AB - Monoclonal gammopathies (GM) are divided into clinically manifest and asymptomatic syndromes. In the last 20 years, the prevalence of GM in industrialized Countries is more than doubled. In order to verify the prevalence of these forms in Western Sicily, at University General Hospital of Palermo an epidemiologic inquiry was made to check the presence of GM as fortuitous event during routine analyses; from March 2000 to March 2001 it was found a prevalence of 0.89% (196 cases on 22100 analysed). Average age was 67.5, with 58.2% men and 41.8% women from different Departments of University General Hospital. The finding of a monoclonal component must be considered an important laboratory datum to observe with time by not invasive laboratory examinations (high resolution serum-protein electrophoresis and immunofixation). PMID- 12910873 TI - [Anti-HEV seroprevalence in the area of Lecce]. AB - Hepatitis E virus (HEV) is the causative agent of an enterically transmitted viral hepatitis associated with inadequate hygienic conditions. Hepatitis E is endemic in developing countries where it occurs both in sporadic and epidemic form. Outbreaks have been reported in different areas of the world. The aim of the present work was to assess the seroprevalence of HEV infection in the general population and in different risk groups in the area of Lecce. The presence of anti-HEV antibodies was ascertained by qualitative ELISA (Abbott HEV EIA) and Western Blot (Recomblot HEV IgG/IgM, Mikrogen). The prevalence of anti-HEV in the general population was 2.9% while in intravenous drug users, hemodialized patients and immigrants the seroprevalence rates were 0.7%, 4.3% and 15.3% respectively. Our data show a circulation of HEV at low level in the area of Lecce which is endemic for hepatitis A. Higher seroprevalence rates were found in hemodialized patients and remarkably in immigrants coming from endemic areas like Turkey. PMID- 12910874 TI - Implementation of a surveillance system for stroke based on administrative and clinical data in the Lazio region (Italy): methodological aspects. AB - Stroke is the third leading cause of death and the most important cause of long term disability in Italy and other developed countries, heavily influencing quality of life and costs of health care. In spite of the widespread occurrence of the disease and its relevant impact in Italy, there is neither a national nor a regional surveillance system of cerebrovascular diseases. A regional surveillance system for stroke has two important aims: to help to interpret the geographical and temporal trends of the disease for health care planning and resource allocation and to allow close monitoring of the quality of stroke services. Age-standardized mortality rates for cerebrovascular diseases in the Lazio region (5,242,709 inhabitants) in the period 1998-99 were 69.4 for males and 59.4 for females per 100,000 inhabitants. In the year 2000, about 3% of all hospital discharges were for cerebrovascular diseases with a hospitalisation rate of 4.36 per 1000 inhabitants. The mean length of stay is 12 days (median of 9 days) and in-hospital death is 15.4%. The admission rate for cerebrovascular diseases to emergency departments is 3.40 per 1000 inhabitants. The goal of the Lazio Regional Health Authority is to implement a surveillance system for stroke based both on current data (mortality and discharge data) and on information collected in a registry for quality assessment of stroke care. The first step of the study is to develop a regional register of acute stroke using an 'ad hoc' data sheet integrated in the computer-based patient record system of clinical and administrative data (GIPSE) operating in all emergency departments in the region. PMID- 12910875 TI - [Criteria for the management of low back pain in health care workers]. AB - Low back pain represents one of the most frequent health problems in several working populations and is an important problem for occupation health doctors. In fact, these professionals are often required to face with the specific management of this condition, also by participating with other professionals into the implementation of organizational and structural measures. This contribute describes a variety of interventions aimed at treating the syndrome and recovering the working ability of the patient-worker, including the initial clinical evaluation, the assessment of the casual relationship between work and the syndrome, the treatment, the formulation of proposals to modify the worker's physical and working activity, the ongoing heath surveillance and a more in-depth clinical and instrumental evaluation, including surgical treatment. PMID- 12910876 TI - [Perceived quality of integrated home care in two health districts of L'Aquila]. AB - The present research, as far as its planning and realization is concerned, aims at exploring how ADI (Integrated Home Care) offers its services in two districts of Local Health Unit 04 in L'Aquila; a service that assumes a special relevance in the frame of interventions in favour of the individuals, since it is a valid alternative to hospitalization for disabled citizens or old people having special pathologies. The information collected gives a general outline of the competences involved within ADI, and also of the significance of the results that have been reached out in terms of quality of the assistance, since a subjective measurement, based on indexes of satisfaction, has been used. PMID- 12910877 TI - [Hospital expenditure for the elderly in Italy]. AB - The consequences of demographic and epidemiological transition have deeply involved the health care systems of western countries. Both the highest burden of care and the higher costs of in-patient services are due to the elderly patients. The aim of this paper is to evaluate the percentage of in-patient care due to the elderly patients and the role played by this care on affecting costs. To this purpose all the Italian 1996 discharge reports referred to in-patient care longer than one day, have been analysed. About 40% of in-patient care expenditure is due to over-65 patients while the percentage of the elderly of total Italian population is roughly 17%. The highest age-specific hospitalisation rate are due to cardiovascular and neurological diseases. Only 4 out of 30 most frequent DRGs among elderly patients are surgical pathologies, while the others are medical ones. The implementation of day-hospital care or out-of-hospital care addressed to the elderly will have to take in account these pathologies in order to be effective in both meeting treatment needs and reducing hospital expenditure. PMID- 12910878 TI - [Survey on microclimatic condition of classroom]. AB - The results of a research, in winter and spring, on the microclimatic parameters and lighting in eighteen classrooms of primary schools in the town of Messina, are exposed. In a significant number of classrooms values of temperature and relative humidity are registered not in accordance with Italian standards. Thermal indices (PMV and PPD) showed, in both seasons, discomfort conditions in many rooms. A percentage of dissatisfied persons, significantly higher the 10% (56.8% in winter and 70.3% in spring), has been obtained. We used, also, a questionnaire modified by Fanger to evaluate the thermal subjective sensations of the students. The results point out that the thermal subjective sensations expressed by the students are often in disagreement with the Fanger thermal comfort indices obtained by instrument. The authors believe that the Fanger's indices do not reflect the effective environmental conditions. PMID- 12910879 TI - [Immigration and health: observational study concerning the foreign children attending the Bologna community pediatric service]. AB - An analysis of the health situation of 2583 immigrant children examined by the Community Paediatric Service in the city of Bologna in 1999 and 2000 was made. The data were obtained from health records, from medical notes made in the districts or during school readmissions. Our investigation showed that: 68.8% of the children were of immigrant couples, 21.6% had at least one Italian parent, 6.0% had lost one or both parents and 3.6%, had been adopted by Italian couples; 68.8% were born in EU countries (1620 in Bologna and 133 in other regions). 99.6% of the children had received all compulsory vaccinations; 1853 subjects had also had one or more optional vaccinations: against whooping-cough 45.2%, measles 46.9% and mumps 33.8%. Only 38.5% of children had pathologies, and those most frequently encountered involved the respiratory and digestive systems. Diseases are more often infectious and allergic as in Italian population. Parasitic infections occurred more frequently than in Italian children; amoebiasis and cutaneous diseases were seen in 1.9% of the children (especially Indians). Cases of tuberculosis were limited. The children more often undergoing diagnostic investigations involving hospitalisation were from Asia and Africa. Only 10.1 of mothers and 7.6% of fathers reported medical problems; allergic pathologies were prevalent. PMID- 12910880 TI - [The registry for asbesto-related tumors]. AB - The author stresses the importance of DPCM 10 December 2002 no.308, which determines the format and the rules to fill the registry for the cases of asbestos-related mesothelioma, according to art 36, comma 3, DLgs 277/91. The Author admits the usefulness of such a registry, but comments that its official approval came very late, because it actually was started in 1993 by ISPESL (The Higher Institute for Prevention and Safety of Labor), after the approval of DLgs 277/91. According to ISPESL initiative, all cases of mesothelioma and related circumstances were (and are) collected through a periferal information net operated by COR's. PMID- 12910881 TI - [Studies of respiratory tract diseases in the last 100 years: a chronological record]. PMID- 12910882 TI - [Pathology of respiratory tract diseases]. PMID- 12910884 TI - [Respiratory functions and related diagnostic tests]. PMID- 12910883 TI - [Imaging diagnosis (e.g., thoracic radiography, CT, and MRI)]. PMID- 12910885 TI - [TBLB, VATS/VTLB and BAL--the significance and effects of new technology]. PMID- 12910887 TI - [Pulmonary tuberculosis]. PMID- 12910886 TI - [Surgery of the respiratory organs]. PMID- 12910888 TI - [Infection by atypical acid-fast bacteria]. PMID- 12910889 TI - [Pneumonia and respiratory tract infections]. PMID- 12910890 TI - [Pulmonary mycoses]. PMID- 12910891 TI - [Bronchial asthma]. PMID- 12910892 TI - [Chronic obstructive pulmonary disease (COPD)]. PMID- 12910893 TI - [Interstitial pneumonia (including collagen disease of the lung)]. PMID- 12910894 TI - [Pulmonary sarcoidosis]. PMID- 12910896 TI - [Pulmonary asbestosis]. PMID- 12910895 TI - [Non-asbestos pneumoconiosis]. PMID- 12910897 TI - [Air pollution]. PMID- 12910898 TI - [High-altitude pulmonary edema]. PMID- 12910899 TI - [Pulmonary hypertension]. PMID- 12910900 TI - [Sleep apnea syndrome(SAS)]. PMID- 12910901 TI - [Respiratory insufficiency]. PMID- 12910902 TI - [Lung cancer]. PMID- 12910903 TI - [Mediastinoscopy]. PMID- 12910904 TI - [Development of bronchial fiberscopes]. PMID- 12910905 TI - [Thoracoscopy]. PMID- 12910906 TI - [Selective alveolo-bronchography]. PMID- 12910907 TI - [Astrography]. PMID- 12910908 TI - [Indirect radiography]. PMID- 12910909 TI - [Tsuboi's method of preparing smears for cytodiagnosis]. PMID- 12910910 TI - [Ogawa's method for isolation and culturing mycobacterial organisms]. PMID- 12910911 TI - [Niacin test]. PMID- 12910912 TI - [Pulse oximeter]. PMID- 12910913 TI - [Theory of the initial infection and onset of tuberculosis]. PMID- 12910914 TI - [Experimental creation of a tuberculous cavity]. PMID- 12910915 TI - [Diffuse panbronchiolitis]. PMID- 12910916 TI - [Summer-type hypersensitivity pneumonia]. PMID- 12910917 TI - [HTLV-1-related pulmonary diseases]. PMID- 12910918 TI - [Bronchus-associated lymphoid tissue (BALT)]. PMID- 12910919 TI - [Development of dry BCG]. PMID- 12910920 TI - [Pulmonary surfactant]. PMID- 12910921 TI - [Drugs for the respiratory organs found in the medicine chest belonging to Koan Ogata]. PMID- 12910922 TI - [A list of antitussive agents and expectorants developed in the last 100 years]. PMID- 12910923 TI - [A list of bronchodilator agents developed in the last 100 years]. PMID- 12910924 TI - [A list of anti-hypersensitivity agents developed in the last 100 years]. PMID- 12910925 TI - [A list of non-steroidal anti-inflammatory agents and immunosuppressive agents developed in the last 100 years]. PMID- 12910926 TI - [A list of adrenocortical steroids developed in the last 100 years]. PMID- 12910927 TI - [A list of anti-infective agents developed in the last 100 years]. PMID- 12910928 TI - [A list of anti-neoplastic agents developed in the past 100 years]. PMID- 12910929 TI - [Organizations and facilities that supported the study of the respiratory system in the past 100 years]. PMID- 12910930 TI - [Endocrine profiles and gonadotropin response to Gn-RH of men with testicular cancer]. AB - PURPOSE: To investigate the function of the hypothalamic-pituitary-testicular axis in testicular germ cell tumors, we evaluated gonadotropin responses to gonadotropin-releasing hormone (Gn-RH), semen quality, and serum levels of sex steroid hormones in patients with testicular cancer. PATIENTS AND METHODS: Basal serum levels of luteinizing hormone (LH), follicle stimulating hormone (FSH), and human chorionic gonadotropin-beta (hCG-beta) were measured before and after high orchiectomy in 20 patients with germ cell tumors of the testicle (9 with seminoma and 11 with nonseminomatous tumor). Semen quality and basal serum levels of testosterone, free testosterone, and estradiol were measured before orchiectomy. The Gn-RH test was performed before orchiectomy in all patients and after orchiectomy in patients without detectable gonadotropin levels in pre-operative serum samples. Gonadotropin levels were measured at 0, 30, 60, 90, and 120 minutes after intravenous injection of 100 micrograms of luteinizing hormone releasing hormone (LH-RH). RESULTS: Serum gonadotropin concentrations were not detectable in 6 of 8 (75%) men with hCG positive tumors or in 4 of 12 (33.3%) men with hCG negative tumors before orchiectomy. Before surgery, 10 men without detectable gonadotropin levels showed complete suppression of the LH and FSH responses to LH-RH and 10 men with detectable gonadotropin levels showed significant increases in the LH and FSH responses (p < 0.01) at 30 minutes. After surgery, the Gn-RH test was performed in 9 men without detectable gonadotropin levels prior to surgery. Seven of these 9 men exhibited significant increases in the LH and FSH responses (p < 0.01) at 30 minutes while no response to LH-RH before or after surgery was seen in 2 men with detectable serum hCG-beta. We observed a significantly lower sperm density (median 7.5 x 10(6)/ml, range 0.4 to 17.8) in men with hCG positive tumors than in men with hCG negative tumors (median 33 x 10(6)/ml, range 0 to 103) (p < 0.002). Although testosterone levels did not differ significantly in men with hCG positive tumors and men with hCG negative tumors, free testosterone levels were significantly higher in men with hCG positive tumors (median 28.4 ng/ml, range 8.5 to 39.8) compared with men with hCG negative tumors (median 18.7 ng/ml, range 4.9 to 24.1) (p < 0.002). Estradiol levels were significantly increased in men with hCG positive tumors (median 44 pg/ml, range 26 to 110) compared with men with hCG negative tumors (median 33.5 pg/ml, range 10 to 87) (p = 0.002). CONCLUSION: The present findings indicate that serum hCG producing testicular cancers are associated with a complete suppression of the gonadotropin response to Gn-RH at the pituitary level, resulting in an inhibition of LH and FSH secretion, and also that serum hCG secreted by testicular cancers may suppresses spermatogenesis and may stimulate androgen and estradiol production by the testes. Since suppressed serum gonadotoropin levels are found in men with hCG non-producing testicular cancers, other factors derived from the tumor may cause downregulation of the gonadotropin response to Gn-RH. PMID- 12910931 TI - [Folic acid reduces risks of having fetus affected with neural tube defects: dietary food folate and plasma folate concentration]. AB - OBJECTIVES: Risk of having fetus affected with neural tube defects can be reduced by maternal periconceptional folic acid supplementation. The purpose of the present study is to investigate how folate is taken from diets and to measure plasma folate concentrations. SUBJECTS AND METHODS: A total of 222 women comprising 5 groups, i.e., healthy women, mothers of myelodysplastic patients, pregnant women, myelodysplastic patients, nurse students, participated in our study. Food frequency questionnaires kept 3 days were analyzed based on the 5th standard table of food composition in Japan. Plasma folate concentrations were measured by means of chemiluminescent immunoassay method. Changes in plasma folate concentrations and possible adverse effects following the folic acid supplementation for 16 weeks were also investigated. RESULTS: The dietary intake of folate, plasma folate concentration and energy intake averaged 293 micrograms/day, 8.1 ng/ml and 1,857 Kcal, respectively, among the subjects. Pregnant women took the largest amount of folate from diets and demonstrated the highest plasma folate concentration among the groups. The dietary folate in myelodysplastic patients and nurse students was significantly lower compared to that of healthy women. The Recommended Dietary Allowance of folate was not fulfilled in 22% of non-pregnant adult women and 72% of pregnant women. The dietary folate was mainly taken from the 3rd food group but the 4th group of food was consumed most. Mean folate intake was significantly correlated with circulating concentrations of serum folate (p = 0.012 r = 0.186). The consecutive administration of 400 micrograms supplements for 16 weeks increased a baseline plasma value of 8.7 ng/ml to 32.6 but fell down rapidly to 17.3 24 hours later without any adverse effects. CONCLUSIONS: The dietary folate and serum folate concentrations averaged 293 micrograms/day and 8.1 ng/ml, respectively. The former is the first report based on the 5th standard table of food composition in Japan. Majority of pregnant women took less dietary folate than what recommended by the government. Those who are capable of becoming pregnant are recommended to consume much of the 3rd food group and those who are planning to become pregnant are recommended to take 400 micrograms of folic acid supplements from 4 weeks before to 12 weeks after conception. PMID- 12910932 TI - [Evaluation of psychometric properties of Japanese version of international prostate symptom score and BPH impact index]. AB - PURPOSE: To evaluate psychometric properties of the Japanese version of International Prostate Symptom Score (IPSS) and BPH Impact Index (BII). METHODS: The translated IPSS and BII questionnaires were administered to 103 patients with benign prostatic hyperplasia and 23 asymptomatic men. In 82 patients the questionnaires were repeated 2 weeks later to examine reproducibility. Further 2 weeks later 21 out of 82 patients responded to the questionnaires asking symptoms during the last week rather than during the last month. To evaluated responsiveness the questionnaires were repeated after treatment in 22 patients. Internal consistency, construct validity and criterion validity were examined by proper statistic methods. RESULTS: Reproducibility was good to excellent with weighted kappa more than 0.62 for all items. It was not significantly affected by age, symptom severity, institution type, or whether the patients were asked symptoms during the last week or during the last month. Internal consistency was also good with Cronbach's alpha more than 0.83. Principle component analysis identified 2 components in IPSS and a single component in BII, with all the items contributing to the first component. Most items had significant correlation with external criteria including maximum flow rate, residual urine volume or prostate volume. The scores of patients were clearly larger than asymptomatic men and obviously reduced after receiving therapies. CONCLUSION: Japanese translations of IPSS and BII were shown to be reliable, valid and one-dimensional instruments in the Japanese patients. They would be equivalent to the original English questionnaires. PMID- 12910933 TI - [Mucinous adenocarcinoma of the prostate: a case report and review of 32 cases on immunohistochemical study of both PSA and CEA]. AB - Mucinous adenocarcinoma of the prostate is extremely rare and its biological behavior is not well known. We report a case of mucinous adenocarcinoma of the prostate which stained positively for prostate specific antigen (PSA) and negatively for carcinoembryonic antigen (CEA) on immunohistochemical study. Our case contained conventional adenocarcinomas and no signet-ring cells. Thirty two cases of mucinous adenocarcinoma of the prostate which performed on immunohistochemical study of both PSA and CEA, including our case, were reviewed. 17 of the 23 cases of immunoreactive to PSA contained conventional adenocarcinomas, and 3 of the 10 cases of immunoreactive to CEA contained them, respectively. The 6 cases of immunoreactive to CEA only contained signet-ring cells. It indicated that there seemed to be the two types of mucinous adenocarcinoma of the prostate, the one which stained positively for PSA was the subtype of conventional adenocarcinomas, and the other which stained positively for CEA and negatively for PSA was derived from the intestinal metaplasia with atypia of the prostatic urethra. PMID- 12910934 TI - [A case in which severely irritable bladder following intravesical instillation of Bacillus Calmette Guerin was successfully treated by steroid therapy]. AB - We report a patient whose irritable bladder symptoms following Bacillus Calmette Guerin (BCG) instillation were satisfactorily treated by steroid administration. A 59-year-old male had undergone transurethral resection for the bladder carcinoma recurred three times. The histopathological examination revealed the tumor as transitional cell carcinoma, G1 to G2, and pTa. Subsequently an instillation of 80 mg BCG into the bladder was planned 8 times every 7 days. After the 5th instillation he presented with gross hematuria, painful micturition, pollakisuria, urgency and reduced bladder capacity of 15 ml. The dose was reduced to 40 mg and another 3 instillations were accomplished. Since conventional treatments of anti-cholinergics, analgesics and epidural anesthesia were of little help for the subjective symptoms, he was put on the steroid pulse therapy 2 weeks after completion of the BCG regimen. The treatment gradually improved the subjective symptoms and increased the bladder capacity up to 160 ml. In conclusion, we believe that the steroid pulse therapy deserves considering in the early stage of irritable bladder symptoms following BCG instillation. PMID- 12910935 TI - [A case of post-traumatic high flow priapism]. AB - We report a case of 50-year-old man with high flow priapism after blunt perineal trauma. Patient evaluation included intracavernal blood-gasometry, cavernography, color flow Doppler sonography, and internal pudendal arteriography. Although conservative treatment had been tried by an alpha-adrenergic agent, temporary effect was only obtained. We could successfully treat the priapism by percutaneous embolization of the left internal pudendal artery with autologous blood clot and Gelatin. 3 months after the embolization, erectile function of the patient recovered. High flow priapism as reported to date in the literature is also briefly reviewed. PMID- 12910936 TI - [Laparoscopic adrenalectomy using telementoring system]. AB - We report on our experience in telementoring during laparoscopic adrenalectomy. An experienced laparoscopic surgeon supervised a less experienced surgeon from a control room about 100 meter away. Mentoring was accomplished over a fiber optic cable employing real-time video imaging, two-way audio communication, OES ImageTrac Video System used to control the laparoscopic image, and a telestrator. The patient was a 52-year-old male with primary aldosteronism due to a left adrenal adenoma 2 cm in diameter. The procedure was successfully performed in 195 minutes with minimal blood loss. The patient recovered without complications and returned home on postoperative day 7. As broadband telecommunication expands, telementoring will become an important method for the training and supporting of laparoscopic surgery. PMID- 12910937 TI - [Non-compromised grafting policy in off-pump coronary artery bypass]. AB - BACKGROUND: The off-pump coronary artery bypass (OPCAB) has been prevalent around the world. However the most optimal selection of graft conduits has still been controversial. METHODS: Between January 2002 and April 2003, 126 consecutive isolated coronary artery bypass procedures were attempted to performed without cardiopulmonary bypass. A mean age of the patients was 69.3 (47-90). Fifteen patients were operated on emergently and 16 were urgently, including 15 patients requiring preoperative intra-aortic balloon pumping (IABP). Five patients underwent reoperative coronary bypass. RESULTS: All procedures were completed without cardiopulmonary bypass. The mean number of grafts per patient was 3.23 (range, 1 to 6). The skeletonized arterial conduits were aggressively used [117 left internal mammary artery (LIMA) to 155 sites, 70 right internal mammary artery (RIMA) to 79 sites, 57 right gastro-epiploic artery (RGEA) to 82 sites and 49 saphenous vein (SV) to 91 sites]. Cardiac-related hospital mortality was none. Angiographic assessment of grafts demonstrated that patency of LIMA, RIMA, RGEA and SV were 98.6%, 100%, 100%, and 97.6% respectively. CONCLUSION: OPCAB with skeletonized in situ arterial conduits is secure and feasible. PMID- 12910938 TI - [Off-pump coronary artery bypass grafting using donut and SPY]. AB - Off-pump coronary artery bypass grafting (OPCAB) has been rapidly increased, because of its less invasiveness with low complications. However, graft patency rate highly depends on operators' capability due to technical difficulties. In this article, detail operative procedures are introduced to perform OPCAB in 100% for isolated coronary patients. Selecting better stabilizer may be a key of success. Donut Heart Stabilizer can make a still and stable operative field to anastomose less than 1 mm coronary artery. It is very useful to achieve complete revascularization for all stenosed coronary branches. OPCAB with 9 arterial grafts could be done using Donut. SPY Intra-operative Imaging System is also important to get 100% patency rate of the grafts. Using SPY, we can avoid graft trouble during operation in operation room (OR). SPY image is the best key information for operators to decide revision of the failed grafting. Donut 2 Heart Stabilizer has been improved to make more wide and stable operative field. Donut and SPY is the best combination for OPCAB. PMID- 12910940 TI - [Utility of the perfusion-assist for beating heart coronary artery bypass surgery]. AB - During off-pump coronary artery bypass surgery, concern remains about the possible myocardial injury associated with the transient occlusion and stabilization of the target vessels. To try to minimize myocardial ischemia and achieve hemodynamic stability, we utilized a coronary perfusion catheter combined with the perfusion-assisted direct coronary artery bypass system that enables active and modified coronary perfusion of the target vessel throughout the duration of multiple grafting (modified PADCAB). In the series of 10 patients, perfusion of the target coronary systems averaged 2,072.8 +/- 649.7 ml over 92.9 +/- 26.1 minutes under a constant infusion pressure of 120 mmHg. Nitroglycerin (100 micrograms/l of flow) was delivered directly into the coronary arteries as an additive in all patients. Hemodynamic instability was notably absent in all cases, even in cases that required difficult anastomosis with a relatively long time for the anastomosis. There were no perioperative complications and no detectable myocardial damage (i.e., impairment of myocardial wall motion indicated by echocardiography findings or by > 0.25 ng/ml of troponin-T release compared to the preoperative level) in this series of patients. PMID- 12910939 TI - [Selection of surgical strategy for abdominal aortic aneurysm coexisting with coronary artery disease; one-stage versus two-stage, and off-pump versus on pump]. AB - BACKGROUND: In patients having a combination of severe coronary artery disease (CAD) and expanding abdominal aortic aneurysm (AAA), one-stage operation of simultaneous coronary artery bypass grafting (CABG) and AAA repair has been recommended. METHODS AND RESULTS: Of 96 patients (94 men, average 65 year) with AAA and CAD, 21 patients underwent one-stage operation of simultaneous CABG and AAA repair (Group A-1: off-pump CABG in 10 and on-pump in 11), and 75 underwent two-stage operation of CABG followed by AAA with an interval of less than 6 months in 23 patients (Group A-2) and more than 6 months in 52 patients (Group B). Baseline characteristics were similar between the one-stage and two-stage groups, and off-pump and on-pump subgroups, except the proportion of single vessel disease. Mean number of distal anastomoses was significantly lower in the off-pump subgroup than in the on-pump subgroup (p = 0.011) and the two-stage group (p = 0.0057). There was 1 hospital death among on-pump patients but none among off-pump patients. Mortality rate was 4.8% in the one-stage group. Two patients (8.7 &) in the two-stage group, who had AAA was 7.0 cm and 7.5 cm diameter, died of AAA rupture prior to repair. The 10-year all death free rates were 44% in Group A (A-1 and A-2), and 86% in Group B (p = 0.0004). CONCLUSIONS: Our data suggest that one-stage operation with off-pump CABG is an attractive option in selected patients with a large AAA (> 7 cm). PMID- 12910941 TI - [The surgical technique of heart displacement during off-pump coronary artery bypass grafting in the exposure of the anastomotic site of posterior descending artery]. AB - Heart displacement during off-pump coronary artery bypass grafting (OPCAB) is necessary to expose the anastomotic site. However, such displacement causes hemodynamic deterioration especially in the exposure of the circumflex branches and posterior descending artery (PDA). In this study we present two different cardiac positioning for the exposure of the anastomotic site of PDA. To expose the mid-portion of PDA the heart is vertically displaced by fixing the suction cup of the heart positioner on apex. To expose the proximal portion of PDA the suction cup is attached on the acute margin of the right ventricle (off apex). By using these 2 different techniques properly, hemodynamic deterioration can be avoided during OPCAB for PDA. PMID- 12910942 TI - [Hemodynamic collapse during off-pump coronary artery bypass grafting and optimization]. AB - The present study addressed the incidence of unpredicted hemodynamic collapse in off-pump coronary artery bypass grafting (OPCAB). Since 1999, OPCAB was attempted in 114 patients without preoperative hemodynamic collapse to date. 95% patients of OPCAB were completed without percutaneous cardiopulmonary support (PCPS) and all patients were discharged. In this study, patients are divided into a prophase of 60 patients and an anaphase of 50 patients, in consideration of learning curve. As for the trouble in procedures, 12 patients of ventricular fibrillation, 6 patients of conversion to PCPS, 19 patients of bradycardia which were evaded in temporary pacing were observed. Patients with conversion to PCPS decreased in 5 patients (8%) in prophase and 1 patient (1.8%) in anaphase. Avoidance of hemodynamic collapse were prevention of ischemia by positive use of coronary perfusion, prophylactic intra-aortic balloon pumping (IABP) use for decrease of blood pressure, apical evacuation heart positioner for visual field security. PMID- 12910943 TI - [Routine application of trans-sternal off-pump coronary artery bypass grafting using innovative technique]. AB - OBJECTIVE: With specific apparatus and surgical techniques, we have extended the use of off-pump coronary artery bypass grafting (CABG) [OPCAB] through sternotomy approach to all of the patients with multiple coronary vessel lesions necessitating CABG. The objective of this study is to assess the safety and efficacy of our technique. METHODS: All of the consecutive 100 patients with multiple coronary vessel lesions necessitating CABG underwent trans-sternal OPCAB using our innovative technique between September 2000 and December 2002. Our new technique included left pericardial traction with sternal spreader, right pericardial compression with pericardial retractor and routine use of our original coronary shunt tube. RESULTS: The total number of distal anastomoses was 322 and the individual grafts were to left anterior descending coronary artery branch (LAD) 127, circumflex (CX) 101 and right coronary artery (RCA) 94. The mean number of the grafts was 3.22 +/- 0.97. The conduits included left internal thoracic artery (LITA) 103, right internal thoracic artery (RITA) 29, radial artery (RA) 82, gastro-epiploic artery (GEA) 43 and saphena magna vein graft (SVG) 65. Predischarge graft angiogram showed over all patency of 99.1% (arterial graft 100% and vein graft 95.4%). Postoperative course was uneventful in all cases with no other organ failures. There was no operative or hospital death. CONCLUSIONS: OPCAB of our technique is effective and safe. The early clinical outcome and excellent patency rates suggests its more widespread use in CABG cases. PMID- 12910944 TI - [Current status of off-pump coronary artery bypass grafting in 2003]. AB - Accruing to the development of the apical suction device, the improvement of the suction type stabilizer, and the application of the trapezoidal pericardiotomy, we have performed off-pump coronary artery bypass (OPCAB) as the first choice procedure for coronary artery revascularization. In a recent series of coronary revascularization (100 cases), 99% of coronary artery bypass grafting was performed completely under OPCAB technique. The usage of the bilateral internal thoracic arteries under skeletonized technique with Harmonic Scalpel has become one of the standard procedures even in patients with diabetes mellitus, a high risk for cardiopulmonary bypass. There was no cerebrovascular accident, or mediastinitis in the postoperative course. In case of abdominal aortic aneurysm and arteriosclerosis obliterans with coronary artery disease, OPCAB combined with vascular surgery were performed simultaneously with an acceptable mortality rate (2.8%: 1/35). We believe that OPCAB can be the standard procedure for all patients with coronary artery disease. PMID- 12910945 TI - [Effectiveness and advisability of using a graft in off-pump coronary artery bypass grafting]. AB - In off-pump coronary artery bypass grafting (off-pump CABG: OPCAB) procedures involving multiple vessels, the Aorta No-Touch technique is increasingly being adopted to avoid cerebral infarction, while gastro-epiploic artery (GEA)-left anterior descending coronary artery branch (LAD) anastomosis or composite T grafting is performed to supply blood to multiple coronary arteries through the left internal thoracic artery (LITA) as an inflow conduit. The objective of the present study is to investigate the effectiveness and advisability of grafting in patients who undergo OPCAB involving multiple vessels, including those who undergo standard CABG. CONCLUSIONS 1: Composite T-grafts of the LITA in OPCAB involving multiple vessels can present certain problems with patency, patient survival, and need for subsequent revision procedures. In the event of a graft failure, serious symptoms resembling left main coronary trunk (LMT) may develop. 2: The incidence of cardiac events with GEA-LAD is high, and patency in the distal phase is poor. 3: Ideally, LITA-LAD should be performed alone. 4: Among patients who have been screened before the procedure, satisfactory results are obtained for OPCAB in which the ascending aorta serves as an inflow conduit; this procedure should be considered as a viable option. PMID- 12910946 TI - [Usefulness of semi-skeletonized right gastroepiploic artery]. AB - The right gastroepiploic artery (RGEA) could be harvested easily and safely by using an ultrasonic scalpel. It is easier and faster not to remove a satellite vein from RGEA than to skeletonize it fully. And blood flow of the vein is important, because it is the flow of vasa vasorum of RGEA. Among 70 patients who have bypassed to right coronary arteries (RCA), 25 patients were treated with semi-skeletonized RGEA (SSK-RGEA), the other 45 patients were operated with pedicled RGEA or (if pedicled RGEA was short and small) with the other grafts. An availability of SSK-RGEA for RCA was 100%. And that of pedicled RGEA was 47% (p < 0.001). Twenty-eight patients were operated without a pump. Twenty-three of them (82%) were bypassed with RGEA. In the on-pump cases (42 patients), RGEA were used for 13 (31%) cases (p < 0.05). Early post-operative angiographies revealed 1 occlusion. But the site of occlusion was the origin of RGEA branch from a gastro duodenal artery, and the anastomotic site was patent. This graft was supposed to be occluded post-operatively by arteriosclerosis. Flow competition occurred in two grafts. In conclusion, the SSK-RGEA was useful for RCA bypass grafting. The reliability of RGEA should increase the indication of off-pump coronary artery bypass grafting. PMID- 12910947 TI - [How much can off-pump coronary artery bypass grafting be performed for complete revascularization using only in situ arterial conduits?]. AB - With the rapid advance of percutaneous catheter intervention, the direction taken by surgeons is not only to make coronary artery bypass grafting (CABG) less invasive but also to obtain better long-term results by using more arterial conduits. However, off-pump CABG (OPCAB) with only in situ arterial conduits such as the left and right internal thoracic arteries (ITAs) and the right gastroepiploic artery (RGEA) is technically demanding for the surgeon. The purpose of this study was to demonstrate the feasibility and safety of this technique. From January 2002 to December 2002, 57 patients underwent isolated CABG without cardiopulmonary bypass, excluding minimally invasive direct coronary artery bypass (MIDCAB) cases. There were 44 men and 13 women with a mean age of 64.1 years. Most of the patients (95%) underwent postoperative angiography within 1 month after surgery. The mean number of anastomoses was 3.8 +/- 1.4 (range 2 8). One patient suffered from mediastinitis and another from stroke. All patients were discharged from the hospital. Fifty patients (88%) received only in situ grafts for complete revascularization. Forty-one patients received bilateral ITAs and 27 received bilateral ITAs and RGEA. The mean number of anastomoses using bilateral ITAs and RGEA was 4.5 +/- 1.2 (range 3-8). The patency rate was 98%. Complete arterial revascularization with only in situ arterial conduits was associated with minimal operative risk and a high early graft patency rate, even in the off-pump situation. These results indicate that complete revascularization can be achieved in OPCAB with the exclusive use of in situ arterial conduits in patients with diffuse coronary arterial disease. PMID- 12910949 TI - [Clinical outcome of emergency off-pump coronary artery bypass grafting]. AB - From January 2001 to January 2003, we performed 25 emergency off-pump coronary artery bypass grafting (CABG) procedures for patients with acute myocardial infarction (AMI) or unstable angina pectoris. During the same period, we also performed 2 emergency on-pump beating CABG procedures for patients with left main coronary trunk (LMT) shock syndrome. For the present study, we evaluated the operative results of the 25 cases of emergency or urgent off-pump CABG. The patients were divided into 3 groups, those with acute AMI with cardiogenic shock (group 1; n = 8), acute myocardial infarction without shock (group 2; n = 8), and unstable angina (group 3; n = 9). There were no differences between groups 1 and 2 with regard to age, number of diseased vessels, and preoperative use of an intraaortic balloon pump, however, patients in group 1 had a higher number of completely obstructed coronary arteries. Patients in groups 1 and 2 underwent off pump CABG within 3.5 hours after a coronary angiography or coronary intervention procedure, while those in group 3 underwent off-pump CABG within 2 days of coronary angiography. The mean number of grafts per patient was 1.8, 2.1, and 2.3 in groups 1, 2, and 3, respectively. One group 1 patient with an LMT lesion was transferred to on-pump beating CABG because of hemodynamic instability. The 30 day mortality rate was 38% (3 of 8) in group 1, whereas it was 0% in groups 2 and 3. Intubation time, ICU stay, and postoperative stay were similar among the 3 groups. An early angiographic study was undertaken in all surviving patients and the results demonstrated patency in all of the examined grafts. Although our results are limited, emergency off-pump CABG was found to be safe and feasible for AMI without cardiogenic shock or unstable myocardial ischemia. However, the outcome of this procedure for patients with preoperative cardiogenic shock was not satisfactory, therefore, a combination therapy of appropriate mechanical circulatory support, prior revascularization by catheter intervention, and emergency surgical revascularization are considered to improve survival of those patients. PMID- 12910948 TI - [Sequential bypass grafting with arterial grafts can be functioned? Assessment of evaluation for function of sequential bypass grafting from theoretical model]. AB - As total arterial revascularization in coronary artery bypass grafting (CABG) has been recommended, a sequential bypass technique using arterial grafts has been induced. We evaluate whether a sequential bypass graft can be functioned or not by using a simple simulation model for coronary circulation analogous to the electrical circuit based on Ohm law. The ratio of flow between graft and native coronary artery was determined by the severity of stenosis in the bypassed vessels and the graft diameter. In selection for sequential bypass technique, these factors should be taken into consideration. PMID- 12910950 TI - [Off-pump coronary artery bypass grafting for severe acute coronary syndrome]. AB - Off-pump coronary artery bypass grafting (off-pump CABG: OPCAB) has become a standard procedure, but the indication for the patients with severe acute coronary syndrome (ACS) has not been established. The purpose of this study is to evaluate the role of OPCAB for patients with acute myocardial infarction (AMI) and impending myocardial infarction (IMI). Clinical indication of OPCAB for acute coronary syndrome between November 1997 and December 2002, 14 patients diagnosed ACS out of 220 CABG cases underwent surgery. Twelve male and 2 female with a mean age of 66.3 +/- 7.5 were NYHA grade IV condition before surgery. Three of nine AMI cases and 4 of 5 IMI cases underwent OPCAB. Thirteen cases needed intra aortic balloon pumping (IABP) support pre-operation, in 1 AMI and 2 IMI cases IABP had to be weaned during operation. The mean graft number was 2.6. Except one AMI case with severe cardiac damage, 13 cases were discharged in NYHA grade I-II condition. CONCLUSION: In early onset cases with still rising CK-MB, operative risk and result is difficult to evaluate pre-operatively. Equal results could be seen in IMI cases with no previous myocardial damage. We suggest, that in cases without severe myocardial damage, OPCAB could be one alternative treatment. PMID- 12910951 TI - [Off-pump coronary artery bypass grafting for octogenarians with acute coronary syndrome]. AB - In recent years, experiences with performing off-pump coronary artery bypass (OPCAB) has increased dramatically. Many early reports suggest that this approach may improve outcome by lowering postoperative complications. The benefit of this procedure to elderly patients seems appealing but is not well studied. We sought to review our experience with OPCAB in octogenarians with acute coronary syndrome (ACS) to better define the potential benefit of this approach in this high-risk group of patients. Until March 2003, OPCAB was performed in 21 octogenarians (10 men and 11 women with mean age of 82.9 +/- 2.8 years) with ACS at the department of cardiovascular surgery of National Kanazawa Hospital. Two had myocardial infarction and 3 unstable angina. Mean left ventricular ejection function was 41.2%. 14 patients had a history of previous cerebral infarction. All procedures were completed without hemodynamic deterioration and conversion to on-pump coronary artery bypass grafting (CABG). There was no operative mortality and no occurrence of major complications such as low output syndrome, re-exploration for bleeding, cerebral infarction, perioperative myocardial infarction, and mediastinitis. And no further deterioration of organ function occurred in patients with preexisting central nervous system dysfunction or kidney. Peak CK MB concentrations after surgery were within normal limits. This study demonstrates that CABG can be performed safely on high-risk ACS patients 80 years of age and older without the use of cardiopulmonary bypass. OPCAB may be the operation of choice for octogenarians with ACS requiring surgical myocardial revascularization. PMID- 12910952 TI - [Off-pump versus standard on-pump reoperative coronary artery bypass grafting]. AB - BACKGROUND: Using standard on-pump procedures, repeat coronary artery bypass grafting (re-CABGs) are associated with a higher morbidity and mortality than first-time CABGs. This retrospective study assessed the clinical outcomes of off pump re-CABG versus on-pump re-CABG. METHODS: From 1983, 2nd CABG and 3rd CABGs were performed in 162 patients at our institute. The early and late results of 142 patients who received standard on-pump re-CABGs and 20 patients who received off-pump re-CABGs were evaluated. The 2 groups were similar in age, sex, preoperative ejection fraction (EF) and number of coronary lesions. However, mean number of distal anastomoses per patient was significantly higher in on-pump (2.0 +/- 0.8) than in off-pump re-CABG (1.4 +/- 0.6) [p = 0.0009]. RESULTS: Hospital mortality tended to be higher in the on-pump group (5.6%) than in the off-pump group (0%) [p = 0.6]. Morbidity in term of total postoperative complications tended to be higher for on-pump (14.1%) than in off-pump (0%) [p = 0.14]. The 5 year survival rate tended to be higher in the off-pump group (100%) than in the on-pump group (80.8 +/- 3.5%) [p = 0.096]. CONCLUSIONS: In selected patients, off pump re-CABG can be a reliable and safe option. The mid-term survival and event free survival rates achievable with the off-pump option are encouraging. PMID- 12910954 TI - [Is the quality of off-pump coronary artery bypass grafting equivalent to that of conventional coronary artery bypass grafting]. AB - We analyzed 44 patients undergoing coronary artery bypass grafting (CABG) from November 2001 to March 2003. Of 44 patients, 40 patients (29 men and 11 women; mean age 68.1 +/- 9.2 years) underwent off-pump CABG (OPCAB) [90.9%]. The mean number of bypass grafts was 2.53 +/- 0.78. In-hospital mortality was none and major complications except for one case of reexploration for bleeding were not seen. The early patency rate of over-all grafts was 94.6%. The quality of OPCAB was considered to be equivalent to conventional CABG. PMID- 12910953 TI - [Redo coronary artery bypass operation under beating heart via the left thoracotomy reusing patent grafts]. AB - We report three cases of successful redo coronary artery bypass grafting (CABG) under beating heart via left thoracotomy with recycling of patent grafts previously implanted. Case 1 and 2: Anginal attack relapsed due to occlusion of the proximal portions of saphenous vein grafts (SVG) that had been sequentially anastomosed to the left coronary artery branches. In each case, the distal portion of the SVG was patent and functioned as coronary-coronary bypass. The left internal thoracic artery graft (ITAG) anastomosed to the left anterior descending coronary artery (LAD) was patent. Each patient underwent off-pump CABG through a left posterolateral thoracotomy. CABG was performed with the radial artery to the circumflex coronary artery (case 1) or the circumflex coronary artery and previous SVG (case 2). The proximal ends of radial artery grafts were anastomosed to the descending aorta. Case 3: Simultaneous reoperative CABG and the operation for the pseudoaneurysm that developed after the abdominal aortic graft replacement were performed. Bypass grafting between ITAG and LAD was performed with SVG via a left anterolateral thoracotomy because of severe anastomotic stricture of ITAG-LAD. The postoperative courses was uneventful for all patients. PMID- 12910955 TI - [Benefit of off-pump coronary artery bypass grafting evaluated from the change of the regional myocardial oxygen metabolism during bypass grafting]. AB - Recently, off-pump coronary artery bypass grafting (off-pump CABG: OPCAB) has been considered a safe alternative to conventional CABG (CCAB) for myocardial revascularization, because OPCAB improves operative mortality and morbidity from the view of reduction of multi-organ complications, duration of hospital stay, risks of blood transfusion and operative costs. This study was performed to estimate the benefit of OPCAB by examining the change of myocardial oxygen metabolism during OPCAB. Twenty five patients who had undergone OPCAB including the internal thoracic artery (LITA) on the segment 8 of the left anterior descending coronary artery (LAD) were studied in this series, and divided in 2 groups [17 angina pectoris (AP) cases, 8 old myocardial infarction (OMI) cases]. With 3-wave length near infrared spectroscopy (TOS-96: TOSTEC, Tokyo, Japan), the tissue hemoglobin (Hb) volume in the myocardium [Hb index (HbI) = 10 x calibrated absorption at 801 nm/clinical absorption intensity at 801 nm] and the myocardial tissue oxygen saturation (rSo2) on the surface of the left ventricle surrounded by LAD and the 2nd diagonal branch were measured at 5 minutes before and after OPCAB to LAD, and myocardial oxygen metabolism (OM) was calculated [OM = (20-1.34 x systemic Hb x rSo2/100) x HbI]. During OPCAB, several hemodynamic data, systemic Hb and arterial gas findings were not changed. In both groups, the HbI and the OM were significantly increased [HbI: AP group; 1.04 +/- 0.23 to 1.57 +/- 0.41 (p < 0.0001), OMI group; 0.99 +/- 0.28 to 1.55 +/- 0.39 (p = 0.0051), OM: AP group; 11.6 +/- 3.1 to 17.5 +/- 6.0 (p = 0.0010), OMI group; 10.6 +/- 2.3 to 16.1 +/- 2.8 (p = 0.0007)]. The rSo2 was not changed and remained within normal limit during OPCAB. These findings suggested that the regional myocardial oxygen metabolism would be improved immediately just after OPCAB and this finding might be one of the reasons why OPCAB shows the good mortality and morbidity compared with CCAB. PMID- 12910956 TI - [Off-pump coronary artery bypass grafting; its impact on renal function]. AB - PURPOSE: We evaluated the impact of off-pump coronary artery bypass grafting (off pump CABG: OPCAB) on the perioperative renal function. METHODS: Isolated CABG was performed on 359 patients during the period from January 1999 to September 2002. Nine patients on dialysis were excluded from this study and 350 patients were divided into 2 groups: OPCAB Group (n = 214) and on-pump Group (n = 136). Perioperative serum CRE levels of the 2 groups were compared. RESULTS: The ratio of patients with renal impairment (CRE > 1.5 mg/dl) in the OPCAB Group was 8%, which did not differ statistically from that of the on-pump Group (4%). Patients who had renal impairment postoperatively accounted for 20% of the OPCAB Group, which did not differ from that of the on-pump Group (18%). The postoperative CRE/preoperative CRE ratio was lower in the OPCAB Group (1.28) than that of the on-pump Group (1.44). CONCLUSION: The renal function was preserved in the OPCAB Group compared to the on-pump Group. PMID- 12910957 TI - [Procedure and problem for short-term outcomes on off-pump coronary artery bypass grafting]. AB - We performed off-pump coronary artery bypass grafting (CABG) in all cases without reoperation case from July, 2002. Advantage of off-pump CABG versus on-pump CABG which is reduced a number of perioperative complication and early patients recovery was previously demonstrated. In our institute, the mean number of grafts per patients was 4.7 +/- 1.3, and the rate of using arterial grafts was 99.5% in all cases without minimally invasive direct coronary artery bypass (MIDCAB). The mean hospital stay after operation was 10.8 +/- 2.8. It was shorted remarkably in comparison with on-pump CABG; 19.4 +/- 6. Furthermore, sever complication was not occurred in any cases after operation though high risk cases were increased. In the early cases, atrial fibrillation complicated frequently (32%), but using after magnesium sulfate it was remarkably decreased (8.4%). On the other-hand, attention is necessary for the infection caused by the increase of high risk patients. Therefore, we used vancomycin (VCM) at these cases from the viewpoint of prevention. Recently, we performed remnant omental transfer for the sever diabetes mellitus case which was used bilateral internal thoracic artery on CABG. It learned to get the early recovery which was necessary for the off-pump CABG by the above additional treatment. PMID- 12910958 TI - [Economic evaluation of off-pump coronary artery bypass grafting; compared to conventional coronary artery bypass grafting]. AB - This study was undertaken to compare economic outcomes of off-pump coronary artery bypass grafting (off-pump CABG: OPCAB) and conventional CABG (CCABG). We reviewed the medical records of 5 patients with ischemic coronary disease (IHD) who underwent OPCAB and 5 patients with IHD who underwent CCABG. A hospital income which was based on a current Japanese health insurance system was indicated. There were significant differences in total cost (OPCAB = 208,200 +/- 7,383 yen versus CCABG = 324,300 +/- 10,290 yen, 35.8% decreased), costs of medical materials and transfusions (OPCAB = 14,760 +/- 3,270 yen versus CCABG = 87,540 +/- 3,326, 81.3% decreased) and charges of surgery and anesthesia (OPCAB = 130,400 +/- 1,853 yen versus CCABG = 147,650 +/- 4,167 yen, 12% decreased) OPCAB markedly reduces the cost of hospitalization. But charges of surgery and anesthesia of OPCAB seems to be valued properly in a current Japanese health insurance system. PMID- 12910959 TI - [Prognostic factors on surgical treatment for lung cancer]. AB - The factors for determining prognosis after lung cancer surgery are curative operation, pathological stage, pathological classification, adjuvant and neoadjuvant therapies, expression of cancer related genes, and concomitant diseases. In particular, pathological stage (especially N stage) is the most influential factor for tumor recurrence, metastasis and survival. The revision of TNM classification by The Union Internationale Contre le Cancer (UICC) in 1997 was well correlated with prognosis. The 5 year survival rates of stage Ia, Ib, IIa, IIb, and IIIa were 67, 57, 55, 39, and 23%, respectively. Neoadjuvant chemotherapy for clinical N 2 disease is expected to improve surgical curativity and survival. Tumor markers including carcinoembryonic antigen (CEA), CYFRA 21-1, and expression of cancer related genes, such as Ki 67, K-ras, bcl-2 and telomerase, are reported to reflect survival after surgery. Smoking and concomitant pulmonary diseases, such as chronic obstructive pulmonary diseases (COPD) and interstitial lung disease affect postoperative patient quality of life (QOL) and survival. We should examine these factors precisely, predict each patient's prognosis and establish an appropriate treatment strategy for each patient. PMID- 12910960 TI - [Rheumatism and poly(ADP-ribose)]. PMID- 12910961 TI - [Antiphospholipid syndrome: new criteria and management of the patients]. PMID- 12910962 TI - [Multi-center analysis of calcinosis in children with juvenile dermatomyositis]. AB - OBJECTIVES: To reveal the frequency and the clinical characteristics of dystrophic calcification that occurs in children with juvenile dermatomyositis, multi-center analysis was constructed. METHOD: Fifty children with JDM were enrolled, and 14 of them (28.0%) were complicated with calcinosis. Clinical symptoms and laboratory tests at onset, initial therapy and disease course were compared in children with and without calcinosis. RESULTS: The mean age of the onset of calcinosis was 4.78 +/- 3.33 years, and it was younger than those of children without calcinosis (8.66 +/- 3.85 years) (P = 0.0017). No differences of clinical manifestation except Gower's sign were observed. The frequency of positive anti-nuclear antibody was 7.1% in children with calcinosis and 52.9% without calcinosis (P = 0.0112). The initial therapy of methylprednisolon pulses gave no effects on prognosis of calcium deposition. The calcinosis appeared in 1.56 +/- 1.91 year after the onset of the disease. The various types of calcium deposition including large tumorous clumps, subcutaneous plaques or nodules, sheet-type calcification were deserved. They appeared over knee joints (64.3%), elbow joint (64.3%), and hip processes (50.0%). Calcinosis affecting the subcutaneous tissues frequently resulted in painful superficial ulceration of the overlying skin (42.9%), local infection (50.0%), and limitation of joint movement (14.3%). Although aluminum phosphate was effective in 2 children among 7, no other effective treatment was recommended. In 5 cases, surgical removal of tumorous clumps was operated. Thus, juvenile dermatomyositis is frequently complicated with calcinosis. This type of calcinosis was found to be unlikely to resolve completely, and resulted in severe disability in children. PMID- 12910963 TI - [Serum amyloid A as a useful index of disease activity in polymyalgia rheumatica]. AB - PURPOSE: To clarify the usefulness of measuring serum amyloid A (SAA) levels in patients with polymyalgia rheumatica (PMR), we compared this parameter to C reactive protein (CRP). SUBJECTS AND METHODS: The study included 10 patients with PMR, who could be prospectively followed up from the start of prednisolone (PSL) treatment until the CRP level decreased to 1 mg/dl or less. When the CRP level decreased, the subjects were divided into the group in which the symptom persisted (n = 6) and those in which the symptom disappeared (n = 4). RESULTS: In the group in which the symptom persisted, both CRP and SAA levels were significantly higher. When the CRP level decreased, there was no significant difference in the CRP level between the two groups. However, the mean SAA level in the group in which the symptom persisted (137.8 micrograms/ml) was significantly higher than that in the group in which the symptom disappeared (21.8 micrograms/ml). On the initial consultation, there was a positive correlation between CRP and SAA (R = 0.77). The SAA level was more sensitive than the CRP level (y = 94.899 x -51.22). When the CRP level decreased, SAA was much more sensitive (y = 222.92 x +6.9121), suggesting the usefulness of SAA after the start of PSL treatment. CONCLUSION: SAA may be a useful parameter of PMR activity. PMID- 12910964 TI - [The relationship between prognosis after one-day arthroscopic partial synovectomy on rheumatoid knee joints and the histologic features of the synovial membrane]. AB - Forty four rheumatoid arthritis (RA) patients with hydrops of the knee joint had partial synovectomy of the knee joint performed under arthrosopy and subsequent irrigation of the joint with 3,000 ml physiological salile solution. Seventeen out of these 44 cases demonstrated recurring hydrops at one year after the procedures (the recurrence group). The remaider (n = 27) was regarded as the non recurrence group. The histological features consistent with RA (proliferation of surface cells, palisade fasion, multi-nucleated giant cells, lymphocytes, plasma cells and granulation tissue; and the occurrence of fibrinoid necrosis and ferritin depotits) were graded depending on the severity of each feature, according to Koizumi's criteria. The rate of heavy lymphocyte and plasma cell infiltration in the non-recurrence group was significantly higher than that in the recurrence group (P = 0.00 and 0.001, respectively). Therefore, we concluded that the prognosis for one-day arthroscopic partial synovectomy on rheumatoid knee joints is related to the pathological features of the synovial membrane. PMID- 12910965 TI - [Rheumatoid arthritis in patients aged above 75 years old at onset]. AB - Elderly onset rheumatoid arthritis (RA) is difficult to diagnose definitively when the patients note their first symptoms of arthritis above the age of 75 years old. In this report, we reviewed the clinical features of elderly onset RA and its diagnosis. The subjects included 4 females, aged 78, 83, 84 and 93 years, respectively. The onsets were abrupt in 2 cases and more slowly arriving in the other 2. Shoulder joints and wrist joints were involved in all cases. Knee joints, finger joints and foot joints were involved in 3 cases and the elbow was involved in one case. Anti-RA treatment quickly attenuated the acute and severe arthritis and brought down the high CRP level associated with vivid inflammatory activity of RA. The RAPA value was very high in all but one of the cases. Severe destructive findings in radiography was undetectable in the major joints (e.g. shoulder, hip and knee joints). Radiographic findings in wrist and finger joints were also very difficult to differentiate from arthrosis and osteoporosis. On the other hand, RA involvement in the cervical spine was certain in the radiographs; 1 case had anterior atlantoaxial subluxation and 2 cases had subaxial intervertebral erosion. Pathological radiographic findings in the cervical spine are useful for the diagnosis of elderly onset RA. PMID- 12910967 TI - [Pharmacokinetics of salazosulfapyridine in a hemodialysis patient]. AB - The patient was a 62-year-old female. Total gastrectomy was performed due to gastric ulcer in 1969. She was diagnosed as rheumatoid arthritis (RA) in 1985 and was developed to amyloidosis in 1991. She was started on hemodialysis (HD) for chronic renal failure in 1996. In 1998, her arthralgia was aggravated, and 100 mg/day of bucillamine was administered on the day of HD. Her arthralgia persisted, and switching to salazosulfapyridine (SASP) was considered. As there were no standards and no reports for the use of SASP in HD patients, we examined the pharmacokinetics of SASP and its metabolites, and compared our patient with the results of phase one study in normal subjects in Japan. In this case, the blood concentration of SASP was similar to that in healthy controls after single administration of 500 mg of SASP on the day of non-HD, while the concentration of sulfapyridine (SP) was higher than that in healthy donors. However, the blood concentrations of SASP, SP, and N4-acetyl-SP (AcSP) at 24 hours after administration were similar to those obtained in healthy men. SASP was not dialyzed, while about half of SP and AcSP, were dialyzed. In a five-day consecutive administration study also, the blood concentrations of these compounds on Day 5 were similar to those of phase one study, suggesting no accumulation. No adverse drug reaction was observed. As this case had the past history of total gastrectomy and amyloidosis, it is possible that this result is influenced by the factors. Therefore it is necessary to examine pharmacokinetics of SASP and its metabolites beforehand when administering this agent to other HD/RA patients. PMID- 12910966 TI - [A case of primary Sjogren's syndrome with severe pulmonary hypertension and glomerular damage]. AB - We report a case of 58 years old female with primary Sjogren's syndrome who accompanied pulmonary hypertension and glomerular damage. Renal biopsy revealed interstitial nephritis and glomerular damage. Pulmonary perfusion scintigram revealed diffusely decreased pulmonary perfusion, but the defect was not observed. Immunocomplex positive indicated that immune disorder would damage her lung and kidney. Proteinuria and pulmonary hypertension were improved by high dose of prednisolone and low dose of oral cyclophosphamide treatment. No previous reports had shown pulmonary hypertension and glomerular damage complicated with primary Sjogren's syndrome in same patients at same time. But some reports had suggested immune disorder had caused pulmonary hypertension or glomerulonephritis in patients of primary Sjogren's syndrome. Our patient showed immune disorder, and it might cause pulmonary hypertension and glomerular damage. PMID- 12910968 TI - [Adult onset Still's disease associated esophageal cancer: a case report]. AB - We report a case of adult onset Still's disease in a 77 year old man, who was diagnosed as a esophageal cancer 9 months after the onset of this disease. At first malignant lesions in any organs were not found and steroid (prednisolone) therapy was begun and the patient was recovered from manifestations. But while tapering prednisolone to 15 mg, fever, arthritis and rash were observed again. Repeated examination revealed that he had suffered from esophageal cancer. This case is considered as a paraneoplastic syndrome of the esophageal cancer. Patients with adult onset Still's disease should be followed as a paraneoplastic syndrome. PMID- 12910969 TI - [Two cases of rheumatoid arthritis developed after polymyositis]. AB - We report two cases of rheumatoid arthritis (RA) who later had developed after polymyositis (PM). The first patient was 64-year old male who experienced muscular weakness of the four limbs in proximity 10 years ago. He was diagnosed as PM because of the elevated serum CK and the myogenic pattern of EMG, and his symptoms were improved by treatment with corticosteroid. He started to complain polyarthralgia 2 years ago, followed by interstitial pneumonia, pleuritis and skin ulcer. He was admitted because of exacerbated polyarthralgia, multiple subcutaneous nodules, skin eruption and fever. The level of serum CK was within normal range but CRP was elevated and CH 50 was decreased. The laboratory examination showed positive cryoglobulin and high titer of rheumatoid factor, but anti-Jo 1 antibody was negative. The hand X-ray showed bone erosions in bilateral wrist joints. Skin biopsy revealed leukocytoclastic vasculitis. Based on these findings, he was diagnosed as malignant RA. He was successfully treated with methylprednisolone pulse therapy, cyclophosphamide and prostaglandin E 1. The second patient was 77-year old male with pneumoconiosis who experienced muscular weakness of the four limbs in proximity 4 years ago. He was diagnosed as PM based on his clinical and laboratory findings and was treated with temporary corticosteroid. He started to have polyarthralgia last year, and he was admitted because of increasing arthralgia after the treatment of pulmonary tuberculosis. The level of serum CK was slightly elevated due to hypothyroidism, and CRP was highly elevated. Rheumatoid factor and cryoglobulin were positive, but anti-Jo 1 antibody was negative. The hand X-ray showed bone erosions in bilateral wrist joints. Crystals of pyrophosphate calcium was observed in knee joints. He was diagnosed as RA associate with pseudogout. His symptoms were relieved with corticosteroid, salazosulfapyridine and anti-tuberculous therapy. These two cases had altered their clinical features from PM to definite RA, and both had pulmonary complications. Previous reports described the cases of RA followed by PM, most of which were induced by such drugs as D-penicillamine, but the cases of PM who later had developed RA are extremely unusual. The overlapped cases of RA and PM tend to highly associate with pulmonary lesions. PMID- 12910970 TI - [The effects of normothermic cardiopulmonary bypass on plasma concentrations of propofol in arterial, internal jugular bulb, and mixed venous blood]. AB - BACKGROUND: Effects of cardiopulmonary bypass (CPB) on pharmacokinetics of propofol have been investigated based on the analysis using arterial blood concentration (CA). However, it has been indicated that the usual pharmacokinetic analysis may fail to precisely estimate propofol concentrations in the brain and the depth of anesthesia. We have investigated pharmacokinetics of propofol in the brain using internal jugular venous bulb blood (CJ). In the present study, we measured propofol concentrations in the mixed venous blood (CV) as well as CA and CJ to investigate pharmacokinetics of propofol in the brain and other organs. METHODS: Six adult patients undergoing elective coronary artery bypass graft surgery were studied. Blood samples were taken to measure propofol concentrations before, during and after CPB. CPB was performed by nonpulsatile flow at 37 degrees C. RESULTS: CA and CJ decreased significantly immediately after the start of CPB, but soon recovered to the baseline value. On the other hand, CV remained unchanged at a lower level during the study period. Arterio-jugular concentration gradient (CA-CJ) did not change significantly, while arterio-mixed venous concentration gradient (CA-CV) increased significantly during CPB. CONCLUSIONS: Pharmacokinetics of propofol might be different in the brain compared in other organs. PMID- 12910971 TI - [The effect of corticosteroids on postoperative course after esophageal resection for carcinoma]. AB - BACKGROUND: The effect of steroids against inflammatory mediators is well known, but its benefit and adverse effect on the postoperative clinical course are uncertain after esophageal resection for carcinoma. METHODS: Forty-three cases undergoing esophageal resection for carcinoma were studied retrospectively. Twenty-six cases, given corticosteroids during operation or first postoperative day, were classified as the steroid group, and the other 17 cases were defined as the control group. Postoperative courses, complications, and outcomes were compared between these 2 groups. RESULTS: In the steroid group, heart rate, body temperature and C-response protein were depressed during the postoperative period, and duration of systemic inflammatory response syndrome was 1.2 +/- 1.1 days compared with 2.7 +/- 1.4 days in the control group (P < 0.001). Although the duration of mechanical ventilation and ICU stay was unchanged, morbidity rate of postoperative anastomotic leakage was 15% in the steroid group compared with 47% in the control group (P < 0.05). The other morbidity rates of pulmonary complications, postoperative infection, arrhythmia, and liver dysfunction as well as mortality rate were similar between the 2 groups. CONCLUSIONS: Corticosteroids given in the early postoperative period depress systemic inflammatory response and attenuate the rate of postoperative anastomotic leakage after esophageal resection for carcinoma. PMID- 12910972 TI - [Perioperative management of living-donor liver transplantation in two patients with severe portopulmonary hypertension]. AB - Liver transplantation for patients with severe portopulmonary hypertension (PPHTN) has been associated with high mortality. We conducted perioperative management of two patients with severe PPHTN for living-donor liver transplantation. The first case was a 17-year-old male with biliary atresia. He developed dyspnea at the age of 14, for which he was treated with intravenous epoprostenol for 8 months. As a result, the mean pulmonary artery pressure (MPAP) was reduced from 61 to 40 mmHg. Intraoperatively, he was treated with intravenous epoprostenol and nitric oxide (NO) inhalation. His intraoperative course was uneventful but he died from right heart failure on postoperative day (POD) 11. The second case was a 6-year-old girl with biliary atresia. When she was 5 years old, examination for a persistent cough revealed MPAP of 49 mmHg. Neither intravenous epoprostenol nor NO inhalation was effective, and she twice showed transient pulmonary hypertension during the operation. She was extubated 14 hours after the surgery, transferred out of ICU on POD 3 and discharged from the hospital on POD 99. When we compare the two cases, the factors responsible for the success of the management of the second case appear to be early extubation and the short duration of PPHTN. PMID- 12910973 TI - [Perioperative cardiovascular complications in patients with hypertrophic cardiomyopathy]. AB - BACKGROUND: The aim of this study was to examine the incidence of cardiac complications in patients with hypertrophic cardiomyopathy (HCM) during noncardiac surgery. METHODS: A retrospective study was made for surgical patients in the period of 1989-2000 at Kitasato University Hospital. RESULTS: Thirty out of 66000 patients were preoperatively diagnosed as HCM. Sixty percent of the HCM patients had one or more perioperative cardiovascular complications. There were perioperative congestive heart failure in 3 patients (10%), and myocardial ischemia in 4 patients (13%). However, there were no myocardial infarction, no life-threatening dysarrthythmia and no cardiac death. Factors which appeared to be associated with the perioperative cardiovascular complications were the type of HCM (HOCM), major surgery, general anesthesia and preoperative medication with a beta-blocker or a calcium channel blocker. CONCLUSIONS: It is suggested that patients with HCM undergoing noncardiac surgery have a high incidence of cardiac complications such as congestive heart failure and myocardial ischemia. PMID- 12910974 TI - [Use of propofol for painful procedures in cancer pain management]. AB - BACKGROUND: Patients with intractable cancer pain often require non pharmacological analgesic treatment that is accompanied by procedure-related pain. Previous works have shown that propofol infusion in adjunction to regional anesthesia provides appropriate sedation during such painful procedures. However, there are a few reports of its use to reduce procedure-related pain in terminal cancer patients. We report cases of propofol sedation during percutaneous vertebroplasty (PVP) in patients with metastatic vertebral compression fracture. METHODS: Propofol was infused during PVP in eleven cancer patients after obtaining written informed consent. The infusion rate of propofol was adjusted using a target-controlled infusion pump to achieve appropriate sedation levels under monitoring bispectral index of the electroencepharogram. Hepatic and renal functions were evaluated using common serum markers, which were determined using standard hospital laboratory methods. RESULTS: The duration of the procedure was 65.5 +/- 5.5 (mean +/- SD) min. The required infusion rate was 8.66 +/- 1.50 mg.kg-1.hr-1. The interval from the termination of the infusion until emergence was 10.7 +/- 4.2 min. No life-threatening complications or significant changes in liver and renal functions were observed. CONCLUSIONS: Propofol can be used effectively and safely for sedation during PVP in terminal cancer patients. PMID- 12910975 TI - [Postoperative recurrence of venous thromboembolism in patients with episodes of preoperative thromboembolism]. AB - BACKGROUND: Venous thromboembolism (VTE) has become a major health problem in Japan. METHODS: We examined, postoperative recurrence of VTE in patients with preoperative venous thromboembolic events and evaluated factors that are relevant to the occurrence of postoperative VTE. RESULTS: Forty-four cases out of 38500 had been diagnosed preoperatively as having VTE in the period of 1995-2001 at Kitasato University Hospital. Postoperative VTE was detected in four patients (9%) out of these 44 cases. Patients who had thrombophilia (antiphosholipid syndrome) or recent episode of VTE within 1 month were higher in the incidence of postoperative recurrence of VTE than the others. Temporary infra-vena cava filters were placed to prevent pulmonary thromboembolism in twelve patients. In 8 of the patients, thrombi were detected in or at the filter before explantation. Three patients (7%) had undergone thrombolysis therapy. Postoperative VTE was not found in patients who had already received anticoagulant therapy (warfarin) for 3 or more months. CONCLUSIONS: It is concluded that newly developed VTE occurred perioperatively in 9% of patients with preoperative VTE, and that thrombophilia and a recent episode of VTE were considered as risk factors of perioperative VTE. Temporary infra-vena cava filters are a valuable device for prevention of postoperative pulmonary thromboembolism. PMID- 12910976 TI - [Suppression of frequent ventricular fibrillation attacks after induction of anesthesia with fentanyl, sevoflurane and propofol: a case report]. AB - We experienced emergency anesthetic management of a 51-year-old male for exchange of a battery in an implantable cardioverter defibrillator. He had repeated and intractable attacks of ventricular fibrillation (Vf) before induction of anesthesia. Infusion of propofol at 100 mg.hr-1 failed to suppress the Vf attacks in the ICU. However, the Vf attacks were suppressed by the addition of fentanyl 100 micrograms, nitrous oxide and sevoflurane to the infusion of propofol. It is speculated that general anesthetics inhibit Vf attacks by direct suppression of myocardium and reduction of sympathetic nervous activity. PMID- 12910977 TI - [General anesthesia for disabled patients undergoing dental treatment in a general hospital]. AB - BACKGROUND: General anesthesia for dental treatment in disabled patients may present particular problems, especially when it is done in a general hospital, compared with specialized dental hospitals. METHODS: We surveyed 24 patients who underwent dental treatment under general anesthesia in our institution during the last decade. RESULTS: Electrocardiography or chest x-ray photography was not obtained in 8 patients owing to lack of patients' cooperation. Slow induction with sevoflurane was selected in 5 patients, because intravenous cannulae could not be placed owing to their rejection. Nasotracheal intubation was performed in all patients, but no difficulty in intubation was documented. As postoperative complications, we observed convulsion in one patient and muscle rigidity in another, but no critical troubles related to circulatory or respiratory status. In patients who were treated for more than ten teeth, we observed differences in age, body weight, duration of anesthesia, maximum concentration of sevoflurane administered during anesthesia, and use of postoperative analgesics compared with the other patients. CONCLUSIONS: We found particular problems of anesthesia for disable patients undergoing dental treatment in a general hospital, and suggest that information on numbers of teeth to be treated is helpful. PMID- 12910978 TI - [Intrathecal phenol block in a child with cancer pain--a case report]. AB - We present a case of a terminally ill child with cancer pain which was treated with intrathecal phenol block. A 13-year-old boy felt severe pain in the right buttock and leg due to osteosarcoma. Despite treatment with nonsteroidal anti inflammatory drugs, intravenous injection of morphine, continuous infusion of ketamine or continuous lumbar epidural block, his pain increased progressively. Therefore, we performed intrathecal block with 10% phenol glycerine 0.2 ml. After the block, his pain was markedly relieved. He sometimes came back home and joined school events. He did not complain of the pain until his death. We suggest that intrathecal phenol block should be performed in terminally ill children with cancer pain if they do not respond to the usual therapeutic modalities including administration of morphine and continuous epidural block. PMID- 12910979 TI - [Anesthetic management of a patient with squeezed heart by huge sternal tumor]. AB - We experienced perioperative management for excision of a huge sternal chondrosarcoma squeezing the heart. A 46-year-old woman could not sleep due to dyspnea for 6 months. Dyspnea increased in the right decubitus and disappeared in the left decubitus. This suggested that the heart was squeezing the tumor. MRI and echocardiography revealed no invasion to the heart. Surgical removal was proposed. Anesthesia was induced with propofol and fentanyl. As this tumor composed of bone-like tissue and was tightly connected with thoracic cage, we thought that the use of muscle relaxant at the induction of anesthesia would not cause cardiovascular collapse and ventilation insufficiency even if the tumor was huge. Vecuronium was administered to facilitate endotracheal intubation. Hemodynamic variables were stable and mask ventilation was uneventful. Anesthesia was maintained with propofol, sevoflurane, supplemental dose of fentanyl and epidural anesthesia. The tumor had invaded the myocardium and was successfully flaked off from the myocardium without using any assisted devices. The tumor was excised including thoracic wall (sternum and parts of 3rd to 12th ribs, 20 cm in diameter). Thoracoplasty was performed using Marlex Mesh. Muscle relaxant was reversed after the end of the operation. Respiration was stable without flail chest. Endotracheal tube was successfully removed in the operation room. The postoperative course was uneventful. PMID- 12910980 TI - [The use of milrinone in a patient with mitral regurgitation and severe pulmonary hypertension: a case report]. AB - A 65-year-old female with mitral regurgitation was scheduled for the mitral valve surgery. However, because of the severe pulmonary hypertension, the patient needed evaluation on the reversibility of pulmonary hypertension and the safety of the surgical procedure. Milrinone, a phosphodiesterase inhibitor with vasodilating and positive inotropic properties, known to lower pulmonary vascular resistance was administered to the patient. Milrinone showed a prompt decrease of pulmonary artery pressure and an increase of cardiac output, and the cardiac surgery was performed uneventfully. It is considered that milrinone is useful in lowering pulmonary vascular resistance and assessing the reversibility of pulmonary hypertension in patients with severe pulmonary hypertension undergoing cardiac surgery. PMID- 12910981 TI - [A case of excessive hemodilution due to ectopic pregnancy]. AB - A 23-year-old female was admitted to our hospital due to hemorrhagic shock with hemoglobin concentration of 2.4 g.dl-1. A diagnosis of ectopic pregnancy was made and immediate emergency operation was performed with massive blood transfusion and rapid fluid infusion. The postoperative course was uneventful. The reasons for no apparent damage in major organs might be short duration of hemodilution, young age, being a female and immediate operation and blood transfusion. PMID- 12910982 TI - [A case of coronary artery spasm during epidural anesthesia with continuous infusion of propofol]. AB - A 50-year-old male patient was scheduled for left partial pulmonary resection and biopsy. The patient had neither complication nor history of ischemic heart disease. After arriving in the operation room, an epidural catheter was inserted into the epidural space at the T 4-5 intervertebral space. Anesthesia was induced with intravenous propofol 100 mg, fentanyl 100 microgram and vecuronium 6 mg and then a double lumen endotracheal tube was inserted. Anesthesia was maintained with O2 and air (FIO2 0.3-1.0), continuous infusion of propofol, intermittent intravenous administration of fentanyl and epidural injection of 1% lidocaine. Forty-five minutes after the start of operation, ECG showed an elevation of ST segment and soon it passed into ventricular tachycardia and ventricular fibrillation. The patient was treated with cardiopulmonary resuscitation. Fifteen minutes later, ECG returned to sinus rhythm but the elevation of ST segment remained. We considered that these cardiac events were due to coronary spasm, and started continuous infusion of nitroglycerin and nicorandil. One hour later, ST segment returned to normal. The possible inducing factors in this case were altered balance between sympathetic and parasympathetic nervous activity caused by infusion of propofol and epidural block, and alpha-stimulation caused by ephedrine. PMID- 12910983 TI - [Malignant hyperthermia during emergency craniotomy: a case report]. AB - An 18-year-old man underwent emergency craniotomy for acute epidural hematoma under general anesthesia using suxamethonium and isoflurane. Marked rises in end tidal carbon dioxide, body temperature, and serum potassium occurred during the surgery. After intravenous dantrolene sodium administration, the body temperature dropped rapidly. No conspicuous postoperative elevation of serum creatine kinase levels were observed. Postoperative muscle biopsy specimen revealed increased calcium-induced calcium release. PMID- 12910984 TI - [Tension pneumothorax manifested after extubation in a patient who underwent total gastrectomy]. AB - A patient developed tension pneumothorax immediately after extubation. The patient was a 53-year-old man, who underwent total gastrectomy under general anesthesia combined with epidural anesthesia. The posterior mediastinum drainage tube was placed near the site of esophago-jejunum anastomosis. Surgeons reported that they might have injured left diaphragmatic pleura during the procedure. Postoperative chest X-ray showed no abnormal findings in the both lung fields. Patient's trachea was extubated when he emerged from anesthesia. However, Spo2 rapidly dropped from 100 to 88. Re-intubation was performed, and positive pressure ventilation was resumed. The Spo2 returned quickly to 100 without hemodynamic change. Auscultation revealed reduced respiratory sound from the left lung. Diagnosis of tension pneumothorax was made from emergency chest X-ray. Patient's respiration improved when chest tube was inserted, but a large amount of air was continuously drained. Air leakage decreased significantly when the mediastinum drainage tube was tentatively occluded. The possible mechanism of the positive pressure in the thoracic cavity was assumed that air was introduced with spontaneous inspiration from the drainage tube, and damaged pleura played as a check valve. PMID- 12910985 TI - [Intraoperative management of simultaneous biventricular assist device placement in a patient with post-myocarditis dilated cardiomyopathy]. AB - We experienced intra-operative management of a patient with post-myocarditis cardiomyopathy who underwent simultaneous biventricular assist device (BiVAD) placement. Although the BiVAD is considered to replace biventricular pump function, significant discrepancy between the flow of the right ventricular assist device (RVAD) and the left ventricular assist device (LVAD) caused difficulty in post-bypass circulatory management in our case. We administered a vasodilator to decrease pulmonary vascular resistance (PVR) and regurgitant fraction of RVAD flow into right ventricle. Inhaled nitric oxide may provide a favorable decrease of PVR. With BiVAD, comprehensive management including the optimization of systemic blood volume, systemic vascular resistance, PVR and native heart function is essential. Transesophageal echocardiography is useful in intraoperative diagnosis and a guide for decision-making in circulatory management. PMID- 12910986 TI - [Coma caused by hypermagnesemia in a pregnant woman complicated with HELLP syndrome]. AB - This case report describes a pregnant woman with acute renal failure, caused by aggravation of HELLP syndrome, who developed coma induced by overdose of continuous magnesium sulfate administration for eclampsia. Before examination of serum concentration of magnesium, coma was suspected to be the result of brain vascular problem or brain infarction. Patient was treated in ICU and hemodialysis was started, and she recovered fully from abnormal neurological symptoms. This case illustrate that hypermagnesemia should be considered in the pregnant woman with coma at ER and ICU. Furthermore, the accurate information about the continuous administration of magnesium by previous clinician helped us to make correct diagnosis and to treat the patient successfully. PMID- 12910987 TI - [A case of pulmonary artery rupture due to pulmonary artery catheter malposition after the weaning of cardiopulmonary bypass]. AB - An 80-year-old female with aortic regurgitation and angina was scheduled for aortic valve replacement and coronary artery bypass graft. After the induction of anesthesia, central venous catheter and pulmonary artery (PA) catheter were placed into the right internal jugular vein. At the weaning from cardiopulmonary bypass, we noticed pulmonary artery pressure (PAP) on the monitor indicating the PA catheter tip being wedged. Therefore, we pulled the PA catheter until the wedging PAP wave disappeared. However, immediately after repositioning of the PA catheter, massive fresh bleeding was found in the endotracheal tube. We estimated the cause of pulmonary bleeding was PA rupture due to malpositioned PA catheter because surgeons had not manipulated the PA during the operation. Soon after restarting the cardiopulmonary bypass, pulmonary bleeding was stopped. Examination with fiberoptic bronchoscope revealed no further bleeding and weaning from cardiopulmonary bypass was accomplished smoothly. Postoperative course was uneventful. PMID- 12910988 TI - [Useful MRI findings for diagnosis of postspinal headache: a case report]. AB - A 49-year-old man developed severe headache after spinal anesthesia. We found diffuse meningeal enhancement on gadolinium enhanced MRI and diffuse meningeal thickening on plain MRI. These MRI findings and postural headache suggest intracranial hypotension. Injection of autologous blood 10 ml into his epidural space was effective to ameliorate the headache. MRI findings were useful for the diagnosis of postspinal headache. PMID- 12910989 TI - [A questionnaire study about perioperative blood transfusion]. AB - BACKGROUND: Red blood cell concentrates in mannitol-adenine-phosphate, autologous blood transfusion, type & screen (T & S), maximum surgical blood order schedule (MSBOS), and a new guideline have been introduced in blood transfusion. To know the present state of perioperative blood transfusion, we did a questionnaire study. METHODS: A questionnaire was sent to anesthesia departments of 1,600 hospitals. RESULTS: Six hundred and fifty hospitals (40.6%) returned the answers. Informed consent is obtained for blood transfusion in every case in 89.8%. Blood loss volume is the most popular indicator to start blood transfusion. Hemoglobin 8 g.dl-1 and hematocrit 25% are also the indicators to start blood transfusion. As an autologous blood transfusion, predeposited (PAT) is the most commonly used. Even when expected blood loss is 3,000 ml, more than 80% of the respondents do not prepare whole blood. Radiation of blood preparation before transfusion is performed in 89.8%. Only half of the hospitals carry out T & S and/or MSBOS. A new guideline is thought to be inadequate in more than 20%. CONCLUSIONS: A questionnaire study indicates that autologous blood transfusion, radiation, leukocyte depletion, T & S, and MSBOS are introduced in our practice. Whole blood is not used, but a new guideline seems to be inadequate. PMID- 12910990 TI - [New concept of multifocal motor neuropathy and conduction block]. PMID- 12910991 TI - [Charcot-Marie-Tooth disease: an overview]. PMID- 12910992 TI - [The clinical effect of combination therapy with edaravone and sodium ozagrel for acute cerebral infarction]. AB - Sodium ozagrel (ozagrel), a selective thromboxane A2 synthetase inhibitor, has been used for the treatment of various types of acute ischemic stroke, except cardioembolic stroke. Recently, edaravone, a novel free radical scavenger, has been approved for the treatment of acute ischemic stroke within 24 hours after onset. Since these two drugs differ in mode of action, we hypothesized that combination of both drugs would yield further improvement of the outcome of patients with acute ischemic stroke. The clinical efficacy of combination therapy with edaravone and ozagrel for acute ischemic stroke was studied retrospectively, and compared with that of ozagrel alone. A total of 62 patients who suffered acute ischemic stroke within 24 hours after onset during the 10-month period from June 2001 to March 2002, were treated with both edaravone and ozagrel (E-O group), while 76 patients during August 2000 to May 2001, were treated with ozagrel alone (O group). The rate of modified Rankin Scale (MRS) 0 and 1 at discharge in the total ischemic stroke and atherothrombotic stroke, was significantly higher in the E-O group than in the O group. The improvement in MRS also differed between E-O group and O group in total. The difference was significant in patients with atherothrombotic stroke but not in those with lacunar stroke. These results indicate that combination therapy with edaravone and ozagrel is more effective than mono-therapy with ozagrel for the treatment of acute ischemic, especially of atherothrombotic stroke. PMID- 12910994 TI - [An autopsy case of polymyositis with exacerbation of chronic myocarditis]. AB - A 38-year-old man had suffered from general fatigue, mild weakness of proximal muscles, and dry cough in November, 2000. Serum levels of muscle enzymes were elevated. Computed tomography of the chest revealed reticular appearance in the bilateral dorsal lung areas. He did not show any improvement, therefore he was referred to our hospital in April, 2001. He was diagnosed as mild polymyositis with mild interstitial pneumonia. He was treated intravenously with methylprednisolone pulse therapy. During the pulse therapy serum level of creatine kinase was decreased, but he died because of acute pump failure of the heart. The cause of the heart failure could be an exacerbation of chronic myocarditis associated with polymyositis and it was confirmed by autopsy findings. When a patient with mild polymyositis complains of general fatigue, myocarditis should be carefully evaluated because of the high risk of death. PMID- 12910993 TI - [Cheiro-oral syndrome due to a cortical infarction in the precentral gyrus: a case report]. AB - We report a 65-year-old woman with sudden onset of paresthesia on the left side of the lip and left thumb. Neurological examinations did not demonstrate any disturbance of higher brain function or motor function except for subjective thermohyperesthesia of the left thumb. Brain MRI demonstrated a small high intensity lesion on T2-weighted images (T2 WI) and diffusion-weighted images (DWI) in the right anterior precentral gyrus. She was diagnosed with cerebral infarction presenting with pure cheiro-oral syndrome (COS). Her neurological symptoms were completely abolished 4 or 5 days later. The thalamus is the region responsible for COS in the majority of cases while reports that a cortical infarction causes COS are rare, because it is difficult to show a small cortical lesion on CT or conventional T2WI. DWI is superior to T2 WI in discriminating between acute and chronic ischemic lesions as well as in detecting small cortical lesions adjacent to the cerebrospinal fluid. In this patient, the lesion was vaguely hyperintense on T2 WI and difficult to differentiate from an artifact caused by cerebrospinal fluid. The precentral gyrus infarction detected on DWI was thought to cause COS in this patient. Not only lesions of the postcentral gyrus as a primary sensory cortex but also those of the precentral gyrus or opercula causing COS were described in previous reports, indicating that the hand and mouth sensory areas may be widely distributed. Cortical mapping studied by electrical stimulation through subdural grid electrodes also supports this finding. Further examination of the relation between neurological symptoms and localization on MR images such as DWI is needed to clarify the distribution of the sensory cortex. PMID- 12910995 TI - [A patient with multiple system atrophy presenting prolonged levodopa-responsive parkinsonism and off-dystonia of the trunk]. AB - A 46-year-old man had a 7-year history of dopa-responsive parkinsonism. Four years after starting levodopa, he had typical motor complications such as wearing off and peak dose as well as off-period dystonia of his trunk. Brain MRI showed marked atrophy of the brainstem and cerebellum, and the cross sign was present in the pontine base. There was neither abnormal signal intensity nor atrophy in the basal ganglia. Then, he was suspected as having multiple system atrophy (MSA). It is not easy to differentiate MSA from Parkinson diseases, particularly when the patient shows good response to levodopa and motor complications like those seen in Parkinson's disease. If the striatal pathology was not severe and nigral degeneration was prominent, presynaptic parkinsonism might occur in MSA, and putaminal preservation might account for good response to levodopa therapy. In patients with MSA, disproportionate antecollis is common before levodopa treatment, and levodopa induced off-dystonia of his trunk is very rare. PMID- 12910996 TI - [A case of chronic pure motor neuropathy associated with anti-GalNAc-GD1a-IgM antibody]. AB - The patient was a 34-year-old male with chronic pure motor neuropathy with such acute onset as seen in Guillain-Barre syndrome. Neurological symptoms were gradually progressive for 4 weeks, and predominantly noted in the left side. Deep reflexes were normal and the distribution of muscle weakness was uneven. Plasma exchange reduced neurological symptoms. Four weeks later, right drop foot was relapsed. High dose intravenous immunoglobulin was effective. Serial electrophysiological studies indicated the asymmetric reduction of CMAP. Repeated assays of anti-GalNAc-GD1a IgM antibody were positive. This is the first report of chronic pure motor neuropathy as multiple mononeuritric type associated with anti-GalNAc-GD1a IgM antibody. This case adds to our knowledge better understanding of the pathogenetic role of anti-GalNAc-GD1a IgM antibody in inflammatory neuropathies. PMID- 12910997 TI - [Steroid-pulse therapy in Guillain-Barre syndrome associated with cytomegalovirus infection: a case report]. AB - We report a 27-year-old man with Guillain-Barre syndrome (GBS) preceded by cytomegalovirus infection. He was admitted to our hospital because of distal dominant weakness and sensory disturbance 5 days after fever. Double filtration plasmapheresis (DFPP) was performed and clinical symptoms temporary but dramatically improved. However facial nerve palsy, difficulty in swallowing food, weakness, dysautonomia and respiratory failure rapidly progressed within 5 days after the onset. Repeated DFPP failed to improve his symptoms. Two months after the onset, he did not improve at all. On T1-weighted MRI, nerve roots were still enhanced with gadolinium, and CSF examination revealed 1,324 mg/dl of protein. These findings suggested us the existence of continuous inflammation on nerve roots. We gave steroid-pulse therapy. He dramatically improved after this treatment. We repeated steroid-pulse therapy for seven times. He was discharged without any major complication 6 months after the onset. Steroid-pulse therapy should be considered in GBS patients associated with CMV infection when other conventional treatments are ineffective. PMID- 12910998 TI - [Fenestration of the extracranial internal carotid artery: report of two cases]. AB - Fenestration of the internal carotid artery (ICA) is extremely rare on clinical settings. Six cases of fenestration in intracranial and 8 in extracracranial ICA have ever been reported in literature. Authors describe two cases of fenestration at the extracranial ICA that were initially suspected of the dissection of ICA on MRA/I. Both angiography and CT angiography (CTA) disclosed the fenestration, which located at C2 vertebral levels. To confirm the fenestration at extracranial ICA is important in such an occasion that mimic the dissection of ICA, especially in patients who complain of cervical pain, mass and other related symptoms. Differences in the fenestration, the dissection and atherosclerosis on imaging, were reviewed and discussed. PMID- 12910999 TI - [A case of deep cerebral venous thrombosis with sudden convulsion after delivery]. AB - A 25-year-old woman was admitted to our hospital because of convulsions after delivering a baby. Her neurological examination revealed a disturbance of consciousness and weakness in both arms. A brain magnetic resonance imaging scan (MRI) showed thrombosis of the left transverse sinus and many patchy high signals at bilateral basal ganglia and subcortical areas. A digital subtraction angiogram (DSA) of the brain revealed a defect at the left transverse sinus, congestion of the cerebral venous flow at the vein of Galen and vasospastic changes at both posterior cerebral arteries. After treating the patient with a venous infusion of heparin, nicardipine and phenytoin, her neurological deficits improved within a day. When MRI and DSA were repeated 2 weeks later, the abnormal lesions had disappeared. These findings suggest that venous thrombosis and vasospasm had the pathogenesis of eclampsia in this case. PMID- 12911000 TI - [Late onset of spinal epidural abscess after spinal epidural catheterization]. AB - A 76-year-old diabetic woman received epidural catheterization for sigmoid colectomy. Four months later she started to complain of fever and severe lumbago, and finally fell into coma and tetraplegia. She had severe neck stiffness, and lumbar puncture yielded yellowish pus. Methicilin-resistant Staphylococcus aureus (MRSA) was detected in a culture of the epidural pus and blood as a causative organism. Magnetic resonance imaging showed extensive abscess in the posterior epidural space at the level between Th1 and L5. We diagnosed her disease as iatrogenic spinal epidural abscess due to epidural catheterization. After emergent laminectomy and evacuation of abscess, inflammation was gradually improved. She returned to normal except for slight gait disturbance. Spinal epidural abscess often develops rapidly after Staphyloccocus aureus infection. In our case, however, neurological deficits appeared 144 days after insertion of epidural catheter. We must remember that spinal epidural abscess is an important cause of lumbago with high fever, even several months after surgical or catheterial intervention to the spine, for immunocompromised patients with diabetes or neoplasm. PMID- 12911001 TI - [A role of peri-ictal SPECT for the identification of epileptogenic focus]. AB - A 42-year-old man had complex partial epilepsy and secondary generalized seizure, without remarkable abnormalities in interictal EEG and head MRI. Three dimensional SRT (3D SRT) analysis of peri-ictal SPECT data detected hyperperfusion in the right temporal lobe, the basal nucleus and the hippocampus, which showed hypoperfusion during interictal period. These findings suggested that the epileptogenic focus existed in the right temporal lobe. We concluded that 3D SRT analysis of peri-ictal SPECT would be helpful to identify the epileptogenic focus. PMID- 12911002 TI - [Trigeminal neuralgia and neurogenic hypertension due to megadolichobasilar artery anomaly]. PMID- 12911003 TI - [Science in taking medicine]. AB - It is a well-known fact that medicine plays an important role in treatment of several diseases. New development of many drugs led to not only benefits but also unexpected damage to human beings. In Japan, we are plunged into an aging society we have never experienced. We have far more opportunities to take medicine. Under these circumstances, it is very important for each one of us to learn how more reliable to take medicine to prevent unexpected side effects. In this citizen joint symposium, we focused on how to take medicine more logically and effectively from the viewpoint of patients. Participation in this symposium could hopefully help promoting our health care in Hokkaido. PMID- 12911004 TI - Blood storage for forensic hemoglobin analysis using CO-oximeter. AB - Blood samples need to be stored for forensic examination using CO-oximeter in case immediate analysis is not possible at autopsy, or re-examinations are expected. In this study, we examined the condition for storage (temperature and air contact) to keep the initial values of hemoglobin derivatives unchanged for a long period. Fresh venous blood samples were divided into two groups; one group was stored without air and the other group with air headspace above the sample. The samples of each group were stored at 37 degrees C, room temperature (ca. 20 25 degrees C), ca. 4 degrees C and ca. -20 degrees C, respectively. They were analyzed with the use of CO-oximeter (CIBA-CORNING 270) a number of times during 6 weeks. And changes in the values of oxyhemoglobin (O2Hb), methemoglobin (MetHb), carboxyhemoglobin (COHb), deoxyhemoglobin (HHb) and total hemoglobin (THb) were examined. Also, another group of blood samples which were made high in COHb experimentally were stored without air at room temperature, ca. 4 degrees C and ca. -20 degrees C, and changes in the percentages of COHb of these samples were examined during 6 months. Our results indicated the following: When the purpose of storage is preservation of O2Hb, MetHb, COHb or HHb, storage at ca. 4 degrees C is considered appropriate, which can keep initial values for up to 2 weeks. As far as COHb is concerned, longer (at least 6 months) storage is possible at ca. 4 degrees C. And no contact with air is desirable in these storages. For THb, frozen storage may be best, which keeps the initial values for up to at least 4 to 6 weeks. In conclusion, when blood samples are stored for CO oximeter analysis, the temperature and the period of storage should be decided according to the durability of each objective hemoglobin derivative. PMID- 12911005 TI - [A study of functional role of perilipin in lipid metabolism: analysis of perilipin transgenic mice]. PMID- 12911006 TI - [Kinetic analysis of JC virus by real time RT-PCR]. PMID- 12911007 TI - [Pharmacological study on sarcoplasmic reticulum dysfunction associated with phospholamban phosphorylation in diabetic rat myocardium]. PMID- 12911008 TI - [The role of macrophage migration inhibitory factor (MIF) in follicle growth and ovulation]. PMID- 12911009 TI - [Studies of hepatocyte nuclear factor (HNF)-1 alpha, HNF-4 alpha and HNF-1 beta genes in juvenile onset diabetes mellitus without obesity]. PMID- 12911010 TI - [Relationship between histological gastritis and mucosal microvascular observations using magnifying endoscopy]. PMID- 12911011 TI - [Long-term effects of chondroitinase ABC on porcine intervertebral discs]. PMID- 12911012 TI - Collagen type 1 retards tube formation by human microvascular endothelial cells in a fibrin matrix. AB - Angiogenesis, or the formation of new microvessels, is often encountered in pathological situations. A fibrinous exudate can often act as a temporary matrix for the ingrowth of these new microvessels. This matrix consists mainly of fibrin, but is mingled with other plasma components and interstitial collagen fibres. In vitro, capillary-like tube formation can be mimicked by exposing human microvascular endothelial cells (hMVECs), seeded on top of a three-dimensional fibrin matrix, to an angiogenic growth factor (e.g. fibroblast growth factor (FGF)-2) and the cytokine tumour necrosis factor (TNF)-alpha. Plasmin activity is required in this process. We investigated whether the angiogenic potential of hMVECs was altered by the presence of collagen. The addition of type I collagen to fibrin matrices dose-dependently inhibited tube-formation. Tube-formation in these fibrin/collagen matrices by hMVECs required matrix metalloprotease (MMP) activity, as well as plasmin activity. On a pure collagen type I matrix, hMVECs were not able to form tube-like structures in the matrix but formed sprouts. This sprouting required MMP activity and was, in contrast to the tube-like structures in a fibrin matrix, not influenced by hypoxia. These data indicate that the interaction between endothelial cells and different matrix components is of importance for the angiogenic potential of these cells. PMID- 12911013 TI - Pediatric neurology in Poland. PMID- 12911014 TI - [Congenital absence of the inferior vena cava as a risk factor for pulmonar thromboembolism]. AB - The absence of the inferior vena cava is a rare congenital anomaly. Currently its diagnosis is based on non-invasive imaging techniques (computerised axial tomagraphy and nuclear magnetic resonance). In most cases, it constitutes a casual finding upon practising these image tests unrelated to this congenital anomaly. In the symptomatic patients, the complaints associated are secondary to venous insufficiency and/or deep vein thrombosis. Recently the congenital absence of inferior vena cava has been described as a risk factor of deep vein thrombosis in young patients. We present a case of congenital absence of inferior vena cava that was admitted in our hospital because of pulmonary thromboembolism. PMID- 12911015 TI - Protein-deficient diet in pregnant females impairs functional activity of enzyme systems in digestive and non-digestive organs in the offspring. AB - Activity of disaccharidases in various portions of the small intestine markedly increased in 6-month-old offspring of pregnant females feeding a protein deficient diet. Activities of amino- and dipeptidases decreased in the kidneys, while alkaline phosphatase activity remained practically unchanged. Probably, information about protein restriction in nutrition of pregnant females is stored in the biochemical memory of adult offspring and promotes the development of metabolic disturbances. PMID- 12911016 TI - McClellan's FDA: boon to industry, consumers, or both? New FDA chief plans to speed drug approval and reduce costs. PMID- 12911017 TI - NIH director reviews first year on the job. "Roadmap" calls for reorganisation of basic and clinical research. PMID- 12911020 TI - [Stick to the scientific ethics, be sure to avoid impetuosities in medical research]. PMID- 12911019 TI - General practice training. PMID- 12911018 TI - A structured evidence-based review on the meaning of nonorganic physical signs: Waddell signs. AB - STUDY DESIGN: This is a structured, evidence-based review of all available studies addressing the concept of nonorganic findings: Waddell signs (WSs). OBJECTIVES: To determine what evidence, if any, exists for the various interpretations for the presence of WSs on physical examination. SUMMARY OF BACKGROUND DATA: WSs are a group of eight physical findings divided into five categories, the presence of which has been alleged at times to have the following interpretations: Malingering/secondary gain, hysteria, psychological distress, magnified presentation, abnormal illness behavior, abnormal pain behavior, and somatic amplification. At the present time, there is, therefore, significant confusion as to what these findings mean. METHODS: A computer and manual literature search produced 61 studies and case series reports relating to WSs. These references were reviewed in detail, sorted, and placed into tabular form according to the following subject areas: 1) Reliability (test-retest); 2) Reliability (inter-rater); 3) Reliability (factor analysis); 4) Validity, psychological distress; 5) Validity, correlation Minnesota Multiphasic Pain Inventory (MMPI); 6) Validity, correlation abnormal illness behavior; 7) Validity, other behaviors; 8) Validity, as a nonorganic phenomenon; 9) Validity, correlation pain drawing; 10) Validity, functional performance; 11) Validity, treatment outcome; 12) Validity, predicting surgical treatment outcome; 13) Validity, return to work outcome; 14) Validity, secondary gain correlation; and 15) Validity, pain correlation. Each study in each topic area was classified according to the type of study it represented according to the type of evidence guidelines developed by the Agency for Health Care Policy and Research (AHCPR). In addition, a list of 14 study quality criteria was used to measure the quality of each study. Each study was categorized for each criterion as positive, (criterion filled), negative (criterion not filled), or not applicable independently by two of the authors. A percent quality score was obtained for each study by counting the total number of positives obtained, dividing by 14 minus the total number of not applicables, and multiplying by 100. Only studies having a quality score of 75% or greater were used to formulate the conclusions of this review. The strength and consistency of the evidence represented by the remaining studies in each topic area (above) was then categorized according to the strength and consistency AHCPR guidelines. Conclusions of this review for each topic area are based on these results. RESULTS OF DATA SYNTHESIS: Of the 61 studies, four had quality scores below 75% and were not used to generate the results of this review. According to the AHCPR guidelines for strength and consistency of the reviewed data, the following results were obtained: 1) There was consistent evidence for WSs being associated with decreased functional performance, poor nonsurgical treatment outcome, and greater levels of pain; 2) There was generally consistent evidence for WSs not being associated with psychological distress, abnormal illness behavior, or secondary gain; 3) There was also generally consistent evidence that WSs are an organic phenomenon and that they cannot be used to discriminate organic from nonorganic problems; 4) There was inconsistent evidence that WSs do demonstrate inter-rater reliability, do not correlate with the neurotic triad of the MMPI, are associated with poorer surgical treatment outcome, and are associated with nonreturn to work; 5) There was little or no evidence that WSs demonstrate test-retest reliability, or reliable factors, and are associated with self-esteem problems, catastrophizing, or the nonorganic pain drawing. CONCLUSIONS: Based on the above results, the following conclusions were made: 1) WSs do not correlate with psychological distress; 2) WSs do not discriminate organic from nonorganic problems; 3) WSs may represent an organic phenomenon; 4) WSs are associated with poorer treatment outcome; 5) WSs are associated with greater pain levels; 6) WSs are not associated with secondary gain; and 7) As a group, WS studies demonstrate some methodological problems. PMID- 12911021 TI - [Research advances in vancomycin-resistant enterococcal infections]. PMID- 12911022 TI - Bibliography. Current world literature. Genes and nutrition. PMID- 12911023 TI - Bibliography. Current world literature. Nutrition in wasting disease. PMID- 12911024 TI - Bibliography. Current world literature. Carbohydrates. PMID- 12911025 TI - Average out-of-pocket health care costs for Medicare+Choice enrollees increase 10 percent in 2003. PMID- 12911026 TI - The medical malpractice crisis and reform of the tort system. PMID- 12911027 TI - Molecular structure of nucleic acids. A structure for deoxyribose nucleic acid. 1953. PMID- 12911028 TI - Evolutionary conservation and functional synergism of curved DNA at the mouse epsilon- and other globin-gene promoters. AB - Human and mouse globin genes were separated approximately 200 million years ago but still share homology and synergism in many aspects including DNA structure. We first mapped DNA bend sites in the mouse epsilon-globin gene and found that these sites were distributed in a regular manner except in the coding region and their overall average distance was 650.7 bp. The first bend site upstream of the cap site (MepsilonB-1, -334 to -147 bp) was found to contain A + T-rich sequences and features contributing to DNA curvature by computer analysis. Transcription assays using deletion constructs indicated strong promoter activity up to bp -215 in erythriod K562 cells. Therefore, the MepsilonB-1 site was located immediately upstream of the promoter region. A reporter gene assay using a series of constructs containing the promoter region revealed that the MepsilonB-1 site showed repressor activity, and on replacement of the DNA curvature with one from another source the activity was retained. A similar feature was found in the other conserved B-1 sites in the human, bovine, and rabbit beta-like globin genes, with the exception of an unconserved B-1 site in the chicken beta-globin gene. A common feature of these conserved B-1 sites was not the nucleotide sequences but the DNA curvature. Furthermore, a unique nucleosome phase at the MepsilonB-1 site was likely to be directed by DNA curvature. Based on these results, DNA curvature is one of the major features of these promoter regions which might influence transcription through nucleosome positioning. PMID- 12911030 TI - Comparative study of translation termination sites and release factors (RF1 and RF2) in procaryotes. AB - Translation termination is catalyzed by release factors that recognize stop codons. However, previous works have shown that in some bacteria, the termination process also involves bases around stop codons. Recently, Ito et al. analyzed release factors and identified the amino acids therein that recognize stop codons. However, the amino acids that recognize bases around stop codons remain unclear. To identify the candidate amino acids that recognize the bases around stop codons, we aligned the protein sequences of the release factors of various bacteria and searched for amino acids that were conserved specifically in the sequence of bacteria that seemed to regulate translation termination by bases around stop codons. As a result, species having several highly conserved residues in RF1 and RF2 showed positive correlations between their codon usage bias and conservation of the bases around the stop codons. In addition, some of the residues were located very close to the SPF motif, which deciphers stop codons. These results suggest that these conserved amino acids enable the release factors to recognize the bases around the stop codons. PMID- 12911029 TI - Potential for retroposition by old Alu subfamilies. AB - Alu elements sharing sequence characteristics of the "old" subfamilies are thought to currently be retrotranspositionally inactive. We analyzed one of these old subfamilies of Alu elements, Sx, for sequence conservation relative to the consensus and the length of the "A-tail" as parameters to define the presence of potential Alu Sx source genes in the human genome. Sequence identity to the left half or the right half of the Alu Sx consensus sequence was evaluated for 4424 complete elements obtained from the human genome draft sequence. A small subset of Alu Sx left halves were found to be more conserved than any of the Alu Sx right halves. Selection for promoter function in active elements may explain the slightly higher conservation of the left half. In order to determine whether this sequence identity was the result of recent activity, or simply sequence conservation for older elements, PCR amplification of some of the loci containing Sx elements with conserved left/right halves from different primate genomes was carried out. Several of these Sx Alus were found to have amplified at a later evolutionary period (<35 mya) than expected based on previous studies of Sx elements. Analysis of "A-tail" length, a feature correlated with current retroposition activity, varied between Alu Sx element loci in different primates, where the length increased in specific Alu elements in the human genome. The presence of few conserved Alu Sx elements and the dynamic expansion/contraction of the A-tail suggests that some of these older subfamilies may still be active at very low levels or in a few individuals. PMID- 12911031 TI - Evolutionary patterns of codon usage in the chloroplast gene rbcL. AB - In this study we reconstruct the evolution of codon usage bias in the chloroplast gene rbcL using a phylogeny of 92 green-plant taxa. We employ a measure of codon usage bias that accounts for chloroplast genomic nucleotide content, as an attempt to limit plausible explanations for patterns of codon bias evolution to selection- or drift-based processes. This measure uses maximum likelihood-ratio tests to compare the performance of two models, one in which a single codon is overrepresented and one in which two codons are overrepresented. The measure allowed us to analyze both the extent of bias in each lineage and the evolution of codon choice across the phylogeny. Despite predictions based primarily on the low G + C content of the chloroplast and the high functional importance of rbcL, we found large differences in the extent of bias, suggesting differential molecular selection that is clade specific. The seed plants and simple leafy liverworts each independently derived a low level of bias in rbcL, perhaps indicating relaxed selectional constraint on molecular changes in the gene. Overrepresentation of a single codon was typically plesiomorphic, and transitions to overrepresentation of two codons occurred commonly across the phylogeny, possibly indicating biochemical selection. The total codon bias in each taxon, when regressed against the total bias of each amino acid, suggested that twofold amino acids play a strong role in inflating the level of codon usage bias in rbcL, despite the fact that twofolds compose a minority of residues in this gene. Those amino acids that contributed most to the total codon usage bias of each taxon are known through amino acid knockout and replacement to be of high functional importance. This suggests that codon usage bias may be constrained by particular amino acids and, thus, may serve as a good predictor of what residues are most important for protein fitness. PMID- 12911032 TI - Codon usage patterns in cytochrome oxidase I across multiple insect orders. AB - Synonymous codon usage bias is determined by a combination of mutational biases, selection at the level of translation, and genetic drift. In a study of mtDNA in insects, we analyzed patterns of codon usage across a phylogeny of 88 insect species spanning 12 orders. We employed a likelihood-based method for estimating levels of codon bias and determining major codon preference that removes the possible effects of genome nucleotide composition bias. Three questions are addressed: (1) How variable are codon bias levels across the phylogeny? (2) How variable are major codon preferences? and (3) Are there phylogenetic constraints on codon bias or preference? There is high variation in the level of codon bias values among the 88 taxa, but few readily apparent phylogenetic patterns. Bias level shifts within the lepidopteran genus Papilio are most likely a result of population size effects. Shifts in major codon preference occur across the tree in all of the amino acids in which there was bias of some level. The vast majority of changes involves double-preference models, however, and shifts between single preferred codons within orders occur only 11 times. These shifts among codons in double-preference models are phylogenetically conservative. PMID- 12911033 TI - A novel strategy for analysis of gene homologues and segmental genome duplications. AB - Transformation-associated recombination (TAR) cloning allows selective isolation of a desired chromosomal region or gene from complex genomes. The method exploits a high level of recombination between homologous DNA sequences during transformation in the yeast Saccharomyces cerevisiae. We investigated the effect of nonhomology on the efficiency of gene capture and found that up to 15% DNA divergence did not prevent efficient gene isolation. Such tolerance to DNA divergence greatly expands the potential applications of TAR cloning for comparative genomics. In this study, we were able to use the technique to isolate nonidentical chromosomal duplications and gene homologues. PMID- 12911034 TI - Perfectly complementary nucleic acid enzymes. AB - The ability to maximize the use of available nucleic acid sequence space would have been crucial during the presumed RNA world and confers selective advantage in many contemporary organisms. One way to access sequence space at a higher density would be to make use of both strands of a duplex nucleic acid for the production of functional molecules. As a demonstration of this possibility, two pairs of nucleic acid enzymes were engineered to be perfect complements, each with the capacity to adopt a distinct structure and catalyze a particular chemical transformation. Both members of each pair of enzymes exhibited nearly the same level of activity as the canonical form of the corresponding catalytic motif. The ability to generate functional nucleic acids encoded by both strands of a duplex has implications for the evolution of catalytic nucleic acids and the prospects for realizing maximum functionality from a given genetic sequence. PMID- 12911035 TI - Epigenetic silencing may aid evolution by gene duplication. AB - Gene duplication is commonly regarded as the main evolutionary path toward the gain of a new function. However, even with gene duplication, there is a loss versus-gain dilemma: most newly born duplicates degrade to pseudogenes, since degenerative mutations are much more frequent than advantageous ones. Thus, something additional seems to be needed to shift the loss versus gain equilibrium toward functional divergence. We suggest that epigenetic silencing of duplicates might play this role in evolution. This study began when we noticed in a previous publication (Lynch M, Conery JS [2000] Science 291:1151-1155) that the frequency of functional young gene duplicates is higher in organisms that have cytosine methylation (H. sapiens, M. musculus, and A. thaliana) than in organisms that do not have methylated genomes (S. cerevisiae, D. melanogaster, and C. elegans). We find that genome data analysis confirms the likelihood of much more efficient functional divergence of gene duplicates in mammals and plants than in yeast, nematode, and fly. We have also extended the classic model of gene duplication, in which newly duplicated genes have exactly the same expression pattern, to the case when they are epigenetically silenced in a tissue- and/or developmental stage-complementary manner. This exposes each of the duplicates to negative selection, thus protecting from "pseudogenization." Our analysis indicates that this kind of silencing (i) enhances evolution of duplicated genes to new functions, particularly in small populations, (ii) is quite consistent with the subfunctionalization model when degenerative but complementary mutations affect different subfunctions of the gene, and (iii) furthermore, may actually cooperate with the DDC (duplication-degeneration-complementation) process. PMID- 12911036 TI - Analysis of microsatellites in 13 hemiascomycetous yeast species: mechanisms involved in genome dynamics. AB - We have analyzed all di-, tri-, and tetranucleotide repeats in the partially sequenced genomes of 13 hemiascomycetous yeast species, and compared their sequences, lengths, and distributions to those observed in the genome of Saccharomyces cerevisiae. We found that most of the 13 species exhibit a unique distribution of microsatellites, not correlated to the base composition of their genome. Species close to S. cerevisiae exhibit a similar distribution, while species more distantly related show a more divergent distribution. We propose that de novo formation and continuous loss of microsatellites are active processes generating new DNA sequences. We also show that hemiascomycete-specific genes encoding transcription factors contain trinucleotide repeats more frequently than expected from their average frequency distribution. These transcription factors might play an important role in the speciation process, by regulating gene expression through DNA-protein or protein-protein interactions mediated by stretches of charged amino acids encoded by trinucleotide repeats. PMID- 12911038 TI - Neutral evolution of ten types of mariner transposons in the genomes of Caenorhabditis elegans and Caenorhabditis briggsae. AB - Ten types of mariner transposable elements (232 individual sequences) are present in the completed genomic DNA sequence of Caenorhabditis elegans and the partial sequence of Caenorhabditis briggsae. We analyze these replicated instances of mariner evolution and find that elements of a type have evolved within their genomes under no selection on their transposase genes. Seven of the ten reconstructed ancestral mariners carry defective transposase genes. Selection has acted during the divergence of some ancestral elements. The neutrally-evolving mariners are used to analyze the pattern of molecular evolution in Caenorhabditis. There is a significant mutational bias against transversions and significant variation in rates of change across sites. Deletions accumulate at a rate of 0.034 events/bp per substitution/site, with an average size of 166 bp (173 gaps observed). Deletions appear to obliterate preexisting deletions over time, creating larger gaps. Insertions accumulate at a rate of 0.019 events/bp per substitution/site, with an average size of 151 bp (61 events). Although the rate of deletion is lower than most estimates in other species, the large size of deletions causes rapid elimination of neutral DNA: a mariner's "half-life" (the time by which half an element's sequence should have been deleted) is approximately 0.1 subsitutions/site. This high rate of DNA deletion may explain the compact nature of the nematode genome. PMID- 12911037 TI - Phylogenetic analysis of plant basic helix-loop-helix proteins. AB - The basic helix-loop-helix (bHLH) family of proteins is a group of functionally diverse transcription factors found in both plants and animals. These proteins evolved early in eukaryotic cells before the split of animals and plants, but appear to function in 'plant-specific' or 'animal-specific' processes. In animals bHLH proteins are involved in regulation of a wide variety of essential developmental processes. On the contrary, bHLH proteins have not been extensively studied in plants. Those that have been characterized function in anthocyanin biosynthesis, phytochrome signaling, globulin expression, fruit dehiscence, carpel and epidermal development. We have identified 118 different bHLH genes in the completely sequenced Arabidopsis thaliana genome and 131 bHLH genes in the rice genome. Here we report a phylogenetic analysis of these genes, including 46 genes from other plant species and a classification of these proteins into 15 distinct plant clades. Results imply a polyphyletic origin for the plant bHLH proteins related only by their bHLH DNA binding motif. We suggest that plant bHLH proteins are under weaker selective constraints than their animal counterparts and that lineage specific expansions and subfunctionalization have fashioned regulatory proteins for plant specific functions. PMID- 12911039 TI - A case for the extreme antiquity of recombination. AB - Recombination is usually assumed to be a mode of reproduction that evolved long after asexual reproduction in response to specific genetic and environmental circumstances. Here the argument is made that recombination was an evolutionary development as ancient as the origins of life. To support this proposition four lines of evidence are given, in particular, the need for primordial genomes to acquire substantial length and to escape from Muller's Ratchet. PMID- 12911040 TI - Allogeneic stem cell transplantation for chronic myeloproliferative disorders and myelodysplastic syndromes: the question is "when?". PMID- 12911041 TI - A critical evaluation of Internet marketing of products that contain ephedra. AB - OBJECTIVE: To evaluate information contained within Internet sites that advertise and market dietary supplements containing ephedra. MATERIAL AND METHODS: We conducted an Internet search to identify Web sites advertising weight-loss supplements that contained ephedra. Between July 7 and July 18, 2002, 4 search engines were used by entering the term herbal weight loss. Outcome measures included disclosure of potential adverse effects of or contraindications to ephedra-containing supplements, disclosure of ephedra alkaloid dosage, and presence of misleading or incorrect information. RESULTS: Thirty-two products and advertisements were identified and systematically evaluated for deviance from truth-in-advertising standards. Of the 32 Web sites analyzed, 13 (41%) failed to disclose potential adverse effects or contraindications to supplement use. Seventeen (53%) did not reveal the dosage of ephedra alkaloids that was recommended. More importantly, 11 sites (34%) contained incorrect or misleading statements, some of which could directly result in serious harm to consumers. CONCLUSION: If dietary supplements containing ephedra are to continue to be marketed freely, substantial reform in advertising regulation and enforcement is warranted. PMID- 12911042 TI - The role of autopsy in the intensive care unit. AB - OBJECTIVE: To identify the frequency and spectrum of clinically relevant diagnoses found at autopsy but not determined before death in adult patients admitted to an intensive care unit (ICU). PATIENTS AND METHODS: We retrospectively reviewed medical records and autopsy reports of patients admitted to ICUs from January 1, 1998, to December 31, 2000. Disagreements between autopsy and antemortem diagnoses were classified as type I or type II errors. A new major diagnosis with potential for directly impacting therapy was considered a type I error. Type II errors included important findings that would not have likely changed therapy. RESULTS: Of 1597 deaths in all ICUs during the study period, autopsies were performed in 527 patients (33%). Autopsy reports were available in 455 patients, of whom 19 (4%) had type I errors and 78 (17%) had type II errors. The most common type I error was cardiac tamponade. There were no significant differences in age, sex, or length of stay in the ICU or hospital among patients with and without diagnostic errors or among patients with type I and II errors. Seventy-eight patients had 81 type II errors. Organ transplant recipients had more type I or II errors than did nontransplant patients (35% vs 20%; P = .04). CONCLUSIONS: Diagnoses with impact on therapy and outcome are missed in approximately 4% of deaths of adult patients admitted to the ICU. Transplant recipients are especially likely to have occult conditions for which additional therapy might be indicated. PMID- 12911043 TI - Positional change in blood pressure and 8-year risk of hypertension: the CARDIA Study. AB - OBJECTIVE: To assess the relationship between positional blood pressure change and 8-year incidence of hypertension in a biracial cohort of young adults. SUBJECTS AND METHODS: Participants from the Coronary Artery Risk Development in Young Adults (CARDIA) study with complete data from year 2 (1987-1988), year 5 (1990-1991), year 7 (1992-1993), and year 10 (1995-1996) examinations were included (N = 2781). Participants were classified into 3 groups based on their year 2 systolic blood pressure response to standing: drop, a decrease in systolic blood pressure of more than 5 mm Hg; same, a change of between -5 and +5 mm Hg; and rise, more than 5-mm Hg increase. RESULTS: The number of participants in each group was as follows: drop, 741; same, 1590; and rise, 450. The 8-year incidence of hypertension was 8.4% in the drop group, 6.8% in the same group, and 12.4% in the rise group (P < .001). Adjusted odds ratios for developing hypertension during the follow-up period in the rise group vs the same group were as follows: in black men, 2.85 (95% confidence interval [CI], 1.43-5.69), in black women, 2.47 (95% CI, 1.19-5.11), in white men, 2.17 (95% CI, 1.00-4.73), and in white women, 4.74 (95% CI, 1.11-20.30). CONCLUSIONS: A greater than 5-mm Hg increase in blood pressure on standing identified a group of young adults at increased risk of developing hypertension within 8 years. These findings support a physiologic link between sympathetic nervous system reactivity and risk of hypertension in young adults. PMID- 12911044 TI - Ethical analysis of withdrawal of pacemaker or implantable cardioverter defibrillator support at the end of life. AB - OBJECTIVE: To describe a series of terminally ill patients who requested (or whose surrogates requested) withdrawal of pacemaker or implantable cardioverter defibrillator (ICD) support and the ethical issues pertaining to these requests. PATIENTS AND METHODS: We performed a retrospective review of the medical records of patients seen at the Mayo Clinic in Rochester, Minn, between January 1996 and June 2002 and identified 6 terminally ill patients who requested (or whose family members requested) withdrawal of pacemaker or ICD support. Potential interventions were an ethics consultation and subsequent withdrawal of pacemaker or ICD support. The study's main outcome measures were death and the context in which it occurred. RESULTS: The mean age of the 6 patients (3 men, 3 women) was 75.5 years. Five had pacemakers, and 1 had an ICD. Five patients had advance directives that indicated a desire to withdraw medical interventions if death was inevitable. Two patients and 4 surrogates requested withdrawal of pacemaker or ICD support. One patient died without withdrawal of support despite an ethics consultation that endorsed its permissibility. Another died while an ethics consultation was in progress. The request to withdraw support was granted in 4 patients, all of whom died within 5 days of withdrawal of support. CONCLUSIONS: Granting terminally ill patients' requests to withdraw unwanted medical support is legal and ethical. Death after withdrawal of support is attributable to the patient's underlying pathology and is not the same as physician-assisted suicide or euthanasia. Clinician familiarity with these concepts may lead to more expeditious withdrawal of unwanted medical support from terminally ill patients. PMID- 12911045 TI - Efficacy and safety of plant stanols and sterols in the management of blood cholesterol levels. AB - Foods with plant stanol or sterol esters lower serum cholesterol levels. We summarize the deliberations of 32 experts on the efficacy and safety of sterols and stanols. A meta-analysis of 41 trials showed that intake of 2 g/d of stanols or sterols reduced low-density lipoprotein (LDL) by 10%; higher intakes added little. Efficacy is similar for sterols and stanols, but the food form may substantially affect LDL reduction. Effects are additive with diet or drug interventions: eating foods low in saturated fat and cholesterol and high in stanols or sterols can reduce LDL by 20%; adding sterols or stanols to statin medication is more effective than doubling the statin dose. A meta-analysis of 10 to 15 trials per vitamin showed that plasma levels of vitamins A and D are not affected by stanols or sterols. Alpha carotene, lycopene, and vitamin E levels remained stable relative to their carrier molecule, LDL. Beta carotene levels declined, but adverse health outcomes were not expected. Sterol-enriched foods increased plasma sterol levels, and workshop participants discussed whether this would increase risk, in view of the marked increase of atherosclerosis in patients with homozygous phytosterolemia. This risk is believed to be largely hypothetical, and any increase due to the small increase in plasma plant sterols may be more than offset by the decrease in plasma LDL. There are insufficient data to suggest that plant stanols or sterols either prevent or promote colon carcinogenesis. Safety of sterols and stanols is being monitored by follow-up of samples from the general population; however, the power of such studies to pick up infrequent increases in common diseases, if any exist, is limited. A trial with clinical outcomes probably would not answer remaining questions about infrequent adverse effects. Trials with surrogate end points such as intima-media thickness might corroborate the expected efficacy in reducing atherosclerosis. However, present evidence is sufficient to promote use of sterols and stanols for lowering LDL cholesterol levels in persons at increased risk for coronary heart disease. PMID- 12911046 TI - Hematopoietic cell transplantation for adult patients with myelodysplastic syndromes and myeloproliferative disorders. AB - Hematopoietic cell transplantation (HCT) is currently the only treatment with curative potential for myelodysplastic syndromes (MDS) and myeloproliferative disorders (MPD). Among patients with less advanced MDS, 3-year survival rates of 65% to 75% are achieved with HLA-identical related and unrelated donors. The probability of relapse is less than 5%. Among patients with advanced MDS (> or = 5% marrow blasts), about 35% to 45% who receive transplants from related donors and 25% to 30% who receive transplants from unrelated donors are in remission beyond 3 years. The incidence of posttransplantation relapse is 10% to 35%. Criteria of the International Prognostic Scoring System (originally developed for nontransplant patients) also predict relapse and survival after HCT. Transplantation is successful in 50% to 80% of patients with MPD if performed before leukemic transformation. Depending on the individual risk profile, a considerable number of patients with MDS or MPD are cured by allogeneic HCT. However, HCT should be performed before disease progression. Outcome of patients with treatment-related MDS or with relapse after transplantation remains poor. At present, no definite conclusions can be made with regard to reduced-intensity transplantation regimens. PMID- 12911047 TI - An overview of stem cell research and regulatory issues. AB - Stem cells are noted for their ability to self-renew and differentiate into a variety of cell types. Some stem cells, described as totipotent cells, have tremendous capacity to self-renew and differentiate. Embryonic stem cells have pluripotent capacity, able to form tissues of all 3 germ layers but unable to form an entire live being. Research with embryonic stem cells has enabled investigators to make substantial gains in developmental biology, therapeutic tissue engineering, and reproductive cloning. However, with these remarkable opportunities many ethical challenges arise, which are largely based on concerns for safety, efficacy, resource allocation, and methods of harvesting stem cells. Discussing the moral and legal status of the human embryo is critical to the debate on stem cell ethics. Religious perspectives and political events leading to regulation of stem cell research are presented and discussed, with special attention directed toward the use of embryonic stem cells for therapeutic and reproductive cloning. Adult stem cells were previously thought to have a restricted capacity to differentiate; however, several reports have described their plasticity potential. Furthermore, there have been close ties between the behavior of stem cells and cancer cells. True eradication of cancer will require a deeper understanding of stem cell biology. This article was written to inform medical scientists and practicing clinicians across the spectrum of medical education about the research and regulatory issues affecting the future of stem cell therapy. PMID- 12911048 TI - Stem cell research: elephants in the room. AB - The degree to which these elephants are disruptive to the steady advancement of the adult stem cell field will become clear with time. In some ways they enliven the discourse, but in many ways they interfere with efficient progress. Naming these elephants is a first step toward dealing with them. If we remain aware of these issues when evaluating new research, we are less likely to make careless mistakes, and we are more likely to be able to hold scientists, politicians, journalists, and entrepreneurs accountable for their practices. Although all adult stem cell researchers will spend time profitably riding some of these elephants, we will all inevitably spend more time cleaning up after them. Perhaps open, careful, and unbiased discussions of these elephants will help the cleanup work be less odious and completed sooner, rather than later. PMID- 12911049 TI - Stem cell research: why medicine should reject human cloning. PMID- 12911051 TI - Caregiver and clinician shortages in an aging nation. AB - Despite a burgeoning elderly population, the number of nurses and other providers of direct care has declined to critical levels at many health care facilities. There are also insufficient medical clinicians to meet the demand for health services. A shrinking workforce, an aging population, financial pressures, and increased consumer demand will translate into severe personnel deficits in the future. Similarly, family fragmentation and the trend toward bureaucratization of long-term care have reduced the availability of informal caregivers. Younger workers should be considered a scarce resource for health care organization and planning purposes. The ability of the United States to meet its entitlement promises is likely to be compromised by a reduced labor pool and simultaneous budgetary constraints. Because good geriatric care is often labor-intensive, meeting this goal poses an increasing challenge. PMID- 12911050 TI - Bupropion sustained release for treatment of tobacco dependence. AB - Tobacco use is a global pandemic. The most common pharmacological treatments of tobacco use and dependence include nicotine replacement therapy and nonnicotine medications. Bupropion sustained release (SR) is the only first-line nonnicotine medication recommended by the US Public Health Service Clinical Practice Guideline. Randomized controlled clinical trials have shown that bupropion SR doubles abstinence rates compared with placebo. Long-term treatment with bupropion SR may reduce or delay smoking relapse. Bupropion SR has an excellent adverse effect profile, although a risk exists for serious adverse effects such as seizures. The risk of serious adverse effects associated with bupropion SR can be reduced by careful selection of patients. This article reviews the evidence of efficacy and common adverse effects of bupropion SR and delineates the clinical characteristics of patients at higher risk for adverse effects when bupropion SR is prescribed for treatment of tobacco use and dependence. PMID- 12911052 TI - 23-year-old woman with diffuse muscle and joint pain. PMID- 12911053 TI - Mayo Health System: a decade of achievement. PMID- 12911054 TI - Cutaneous T-cell lymphoma. PMID- 12911055 TI - Systemic herpes-like virus in catfish Ictalurus melas (Italy) differs from Ictalurid herpesvirus 1 (North America). AB - A herpesvirus was isolated during 2 occurrences of mass mortality among adult catfish Ictalurus melas raised in different farms in northern Italy. The agent replicated in the channel catfish ovary (CCO) cell line from channel catfish I. punctatus, inducing a cytopathic effect similar to that caused by Ictalurid herpesvirus 1 (also referred to as channel catfish herpesvirus, CCV). The new herpesvirus, designated I. melas herpesvirus (IcmHV) did not react with polyclonal rabbit or monoclonal antibodies directed to CCV in either neutralization or indirect immunofluorescence assays. The virions of IcmHV possessed a hexagonal nucleocapsid of 107 nm in diameter surrounded by an envelope with a diameter of 227 nm (n = 20) typical for members of the family Herpesviridae. Virions of IcmHV purified from infected CCO cells contained 17 polypeptides ranging in size from 17.5 to 175 kDa and most differed in molecular weight from those found for CCV. The IcmHV was also distinct from CCV when compared by restriction fragment length polymorphisms (RFLP) of genomic DNA following digestions with the endonucleases Kpn I and Sac I. Lastly, the virulence of IcmHV for channel catfish fry and juveniles, respectively, was demonstrated by experimental infections induced by bath exposure or intraperitoneal injection that resulted in 78 to 96% cumulative mortality in groups of exposed fish. Preventing the introduction of this agent into geographic regions where significant channel catfish production occurs should be a high priority. PMID- 12911057 TI - Adhesion of high and low virulence Flavobacterium psychrophilum strains to isolated gill arches of rainbow trout Oncorhynchus mykiss. AB - The ability of Flavobacterium psychrophilum to adhere to the gill tissue of rainbow trout Oncorhynchus mykiss was evaluated. A gill perfusion model was adopted, offering a number of advantages compared to other in vitro as well as in vivo models. A comparison between the adhesion capacity of a high and low virulence F. psychrophilum strain was made. Experiments were additionally carried out to assess the influence of water quality (organic material, nitrite) and temperature on the adhesion process of the bacterial cells. The high virulence strain attached more readily to the gill tissue than did the low virulence strain. Moreover, the adherence of the high virulence strain of F. psychrophilum was influenced by a number of factors. These were immersion of the gill arches in water to which organic material or nitrite were added, and elevated temperature. The former 2 increased the adhesion ability, while the latter had a negative influence on the adherence process. PMID- 12911056 TI - Nanoinjection as a tool to mimic vertical transmission of Flavobacterium psychrophilum in rainbow trout Oncorhynchus mykiss. AB - Newly fertilised eggs of rainbow trout Oncorhynchus mykiss were nanoinjected with Flavobacterium psychrophilum in order to mimic vertical transmission. Two bacterial isolates with different elastin-degrading capacity were used. All infected groups (10, 100 and 1000 colony forming units egg(-1)) showed significantly higher cumulative mortalities than the control groups at the end of the experiment, 70 d post-hatching. The total mortalities in the control groups were below 2.5%. In the high-dose groups, 95 to 100% of the eggs died during the eyed stage. In the intermediate group infected with the elastin-negative isolate, the major mortality occurred during the eyed stage of the egg, with a total cumulative mortality of 83% at the end of the experiment. In the intermediate group infected with the elastin-positive isolate, a total mortality of 63% was recorded. In this group, diseased fry showed clinical signs of disease and morphological changes similar to those described in connection with rainbow trout fry syndrome (RTFS) shortly after the beginning of feeding. In the low dose groups, the mortality in the elastin-negative group was 14% and in the elastin positive group 11%. The bacterium was isolated from dead eggs and fry in infected groups and demonstrated in internal organs of dead and moribund fry by immunohistochemistry. The nanoinjection method used in this study may be a useful method to study pathogens, like F. psychrophilum, that can be vertically transmitted. PMID- 12911058 TI - Development of a PCR-based method for the detection of Listonella anguillarum in fish tissues and blood samples. AB - A PCR assay for detection and identification of the fish pathogen Listonella anguillarum was developed. Primers amplifying a 519 bp internal fragment of the L. anguillarum rpoN gene, which codes for the factor sigma54, were utilized. The detection limit of the PCR using L. anguillarum pure cultures was approximately 1 to 10 bacterial cells per reaction. For tissue or blood samples of infected turbot Scophthalmus maximus, the detection limit was 10 to 100 L. anguillarum cells per reaction, which corresponds to 2 x 10(3) to 2 x 10(4) cells g(-1) fish tissue. Our results suggest that this PCR protocol is a sensitive and specific molecular method for the detection of the fish pathogen L. anguillarum. PMID- 12911059 TI - Molecular cloning, expression of orange-spotted grouper goose-type lysozyme cDNA, and lytic activity of its recombinant protein. AB - Lysozyme acts as a non-specific innate-immunity molecule against the invasion of bacteria pathogens. A leukocyte cDNA library of orange-spotted grouper Epinephelus coioides was constructed and the goose-type (g-type) lysozyme cDNA was isolated. The complete cDNA consists of an open reading frame of 585 bp encoding a protein of 194 amino acids. This protein shows a 72.2% amino acid sequence identity with the flounder g-type lysozyme. Similar to most other species, the glu catalytic residue in g-type lysozymes of the grouper is conserved. Furthermore, like the flounder and carp, the 4 conserved cysteine residues identified in avian and mammalian g-type lysozymes were also absent from the grouper. Northern blot analysis indicated that the g-type lysozyme was expressed in intestine, liver, spleen, anterior kidney, posterior kidney, heart, gill, muscle and leukocytes. In addition, RT-PCR analysis detected the g-type lysozyme transcripts in the stomach, brain and ovary. When an orange-spotted grouper was injected with Vibrio alginolyticus, the number of lysozyme mRNA transcripts detected in the stomach, spleen, anterior kidney, posterior kidney, heart, brain and leucocytes increased 72 h after injection. Recombinant grouper g type lysozyme produced in the Escherichia coli expression system showed lytic activity against Micrococcus lysodeikticus, V. alginolyticus from Epinephelus fario, V. vulnificus from culture water, Aeromonas hydrophila from soft-shell turtle, A. hydrophila from goldfish and V. parahaemolyticus, Pseudomonas fluorescens and V. fluvialis from culture water. PMID- 12911060 TI - Infection patterns of Myxobolus heterospora in two tilapia species (Teleostei: Cichlidae) and its potential effects. AB - A Myxobolus heterospora (Baker, 1963) infection was found in 2 euryhaline tilapia species, Sarotherodon melanotheron melanotheron (Ruppel, 1853) and Tilapia zillii (Gervais, 1852), from a brackish water lake, Lake Nokoue (Benin, West Africa). The histology and ultrastructure of different levels of infection in intestinal connective tissues and wall tissues is described. A total of 391 S. melanotheron melanotheron and 222 T. zillii were examined from October 1987 to October 1989. M. heterospora was found throughout the study period, with a total prevalence of 42.19 and 26.57% for S. melanotheron melanotheron and T. zillii respectively. There was a statistically significant difference in occurrence as a function of season in S. melanotheron melanotheron but not in T. zilli, and there was a significant difference for size and sex in the former and for sex in the latter. M. heterospora induces total destruction of the intestine structure and probably leads to osmoregulatory disturbance. Further investigations of this myxosporean infection are necessary to determine its real effect on the host, since host survival and osmoregulatory rate have not yet been assessed. PMID- 12911061 TI - In vitro studies on viability and proliferation of Enteromyxum scophthalmi (Myxozoa), an enteric parasite of cultured turbot Scophthalmus maximus. AB - In vitro cultivation of the myxozoan Enteromyxum scophthalmi was attempted using different culture media and conditions. The progress of the cultures was monitored using dye-exclusion viability counts, tetrazolium-based cell proliferation assays, measuring the incorporation of BrdU during DNA synthesis, and by morphological studies using light and electron microscopes. In preliminary experiments, the persistence of viable stages for a few days was ascertained in both medium 199 (M199) and in seawater. An apparent initial proliferation was noticed in the culture media, with many young stages observed by Day 7 post inoculation (p.i.). In contrast, fast degeneration occurred in seawater, with but a few living stages persisting to Day 1 p.i and none to Day 5 p.i. Both tetrazolium-based cell-proliferation assays and dye-exclusion viability counts demonstrated a progressive degeneration of the cultures. Although M199 medium and neutral pH with the addition of sera appeared to provide the most favourable conditions during the first few hours, all cultures degenerated with time and no parasite proliferation or maintenance could be achieved in the long term in any of the conditions assayed, including attempts of co-cultivation with a turbot cell line. The ultrastructure of stages cultured for 15 d demonstrated complete degeneration of organelles and mitochondria, although the plasma membrane remained intact in many stages. Unknown factors related to the metabolism or the life cycle of this myxozoan are probably responsible for the inability to culture the parasite, which seems to be strictly dependent on the target host tissues for survival. PMID- 12911062 TI - Trematode metacercariae of fishes as sentinels for a changing limnological environment. AB - Trematode metacercaria populations infecting cichlids in Lake Kinneret (Sea of Galilee) were used as sentinels for the changing limnological environment. Parasitological data from 0+ cichlid fingerlings (Tilapia zillii, Oreochromis aureus, Sarotherodon galilaeus) were collected from the northwest shore of the lake during 1999 to 2001 and compared with data obtained from 1982 to 1984. The results show that the composition of the metacercarial populations changed significantly between the 2 samplings periods. The total number of metacercarial species found in the Kinneret cichlids was lower in 1999 to 2001 than in 1982 to 1984. Metacercariae transmitted by the snail Bulinus truncatus (Clinostomum tilapiae, Euclinostomum heterostomum, Bolbophorus levantinus and Neascus-type metacercariae--black spot and others) that were commonly found in 1982 to 1984 were absent from the fishes sampled during 1999 to 2001. The other trematode metacercariae (Centrocestus sp. and Haplorchis sp. transmitted by Melanoides tuberculata, Pygidiopsis genata transmitted by Melanopsis costata, as well as Phagicola longa, Strigeidae sp.1, an unidentified metacercaria typically found in the liver, and glochidia) increased in abundance in fishes sampled during 1999 to 2001. The calculated 'true' species richness of the habitat, extrapolated as a function of sampling effort, was significantly lower in the 1999 to 2001 versus 1982 to 1984 samples, while significantly fewer fishes needed to be sampled ('sampling effort') during 1999 to 2001 in order to reach the 'true' species richness. The higher calculated values of species richness and diversity for the 1999 to 2001 samples despite the overall decline in species number is explained by the overall increase in metacercariae prevalence. PMID- 12911063 TI - Osmotic cataract causes reduced vision in wild Atlantic salmon postsmolts. AB - Osmotic cataracts were diagnosed in all of 191 Atlantic salmon Salmo salar L. postsmolts caught during 8 trawl hauls on the western side of the Voringsplateau, Norwegian Sea, in June 2001. The changes varied from a hazy opacity in the anterior part of the lens to cataracts affecting the whole lens. Severely affected lenses appeared swollen and large vacuoles were visible in the opaque areas. Large vacuoles in otherwise clear lenses were diagnosed in 1 of 4 adult salmon examined. Histologically, widened sutures, vacuolation of lens epithelium and cortex, and proteinaceous lakes subjacent to the epithelium were the most frequent changes, while extensive cortical necroses and epithelial proliferation were seen in a few cases. UV-absorbance of the aqueous humor was determined and levels compared to plasma levels and also to levels in farmed Atlantic salmon of the same developmental stage. Wild salmon generally showed higher levels of protective factors than farmed fish. The osmotic type of cataract diagnosed leads to poor vision and is a potential cause of reduced survival in postsmolts. The cause of the cataracts could not be determined, but defective osmoregulation is suspected. PMID- 12911065 TI - Enhanced growth and resistance to Vibrio challenge in pond-reared black tiger shrimp Penaeus monodon fed a Bacillus probiotic. AB - The bacterial probiont Bacillus S11 (BS11) was used as a supplement in feed (PF) for black tiger shrimp Penaeus monodon in 2 earthen pond field-trials carried out for 100 d during 2 different seasons in Thailand. Growth and survival were compared with those of shrimp receiving an unsupplemented feed (UF). In the hot and cool seasons, respectively, shrimp fed PF grew significantly larger and had significantly higher survival than shrimp fed UF (p < 0.05). Projected yields on an annual basis (two 100 d crops) were 49% greater with PF-fed shrimp. In 8 d challenge tests using the luminescent bacterium Vibrio harveyi 1526, shrimp fed UF all died within 6 d while survival for shrimp fed PF was 5 and 9%. PMID- 12911064 TI - Metschnikowia bicuspidata and Enterococcus faecium co-infection in the giant freshwater prawn Macrobrachium rosenbergii. AB - In May 2001, an epizootic yeast and bacterial co-infection in the giant freshwater prawn Macrobrachium rosenbergii occurred in Taiwan causing a cumulative mortality of 25%. The diseased prawns had a yellowish-brown body color, milky hemolymph, opaque, whitish muscles, and were approximately 7 mo old with total lengths ranging from 8 to 10 cm. Histopathological examination showed marked edema, yeast infiltration, and necrotic lesions with inflammation in the muscles, hepatopancreas and other internal organs. We isolated 2 pathogens from the diseased prawns, one was a yeast (AOD081MB) and the other a gram-positive coccus (AOD081EF). The gram-positive coccus was identified as Enterococcus faecium by the API 20 Strepsystem, conventional biochemical tests, and it had 99% 16S rDNA sequence identity (GenBank Accession Number AJ276355) to E. faecium (GenBank Accession Number AF529204). The sequence of a PCR product from the D1/D2 domain of 26S rDNA (GenBank Accession Number AF529297) from the yeast gave 99% sequence identity to Metschnikowia bicuspidata (GenBank Accession Number U44822). Experimental infections with these isolates produced gross signs and histopathological changes similar to those observed in the naturally infected prawns. The lethal doses (LD50) for isolate E. faecium AOD081EF, M. bicuspidata AOD081MB and the co-infection were 4.7 x 10(4), 2.6 x 10(2), and 2.4 x 10(2) colony-forming units prawn(-1), respectively. This is the first report of a confirmed co-infection of M. bicuspidata and E. faecium in prawn aquaculture. PMID- 12911066 TI - Physician crying. PMID- 12911067 TI - A tale of two transitions. PMID- 12911068 TI - Obstacles to the delivery of acceptable standards of care in rural home hospices. PMID- 12911069 TI - Baclofen for hiccups. PMID- 12911070 TI - A systematic review on chaplains and community-based clergy in three palliative care journals: 1990-1999. AB - A systematic review of all articles appearing between 1990 and 1999 in the American Journal of Hospice and Palliative Care, the Hospice Journal, and the Journal of Palliative Care was conducted. Articles citing at least one reference were categorized as scholarly, included in the study, and divided into either research or nonresearch categories. Scholarly articles were classified as research if they contained clearly defined methods and results sections, even if these headings were not used. Research and nonresearch articles were subdivided into qualitative and quantitative research and general reviews or program descriptions, respectively. All scholarly articles were read to see if they mentioned clergy, including the terms rabbi, priest, minister, pastor, imam, chaplain, or other religious professionals. Of 838 scholarly articles published between 1990 and 1999 in the three journals, 348 (41.5 percent) were research articles, 417 (49.8 percent) were reviews, and 73 (8.7 percent) were program descriptions. Forty-seven (5.6 percent) of all 838 scholarly articles mentioned clergy or chaplains in some way. Clergy and chaplains were more likely to be an integral part of research articles, whereas mention of them in nonresearch articles tended to be incidental (chi-square = 16.8, p < .001). Moreover, quantitative articles were more likely to include clergy as an integral aspect of the article than were qualitative articles (Fischer's exact probability test, p = .088). The results are discussed with respect to the mutual roles hospice chaplains and community-based clergy play in providing spiritual care at the end of life. PMID- 12911072 TI - Using clinical empowerment to teach ethics and conflict management in antemortem care: a case study. PMID- 12911071 TI - Dying to meet you: facing mortality and enabling patient styles. AB - This paper is intended to emphasize the existence of prognostic uncertainty in providing survival estimates while also providing a method for caring to those who want to authentically help dying patients. Facing one's own mortality helps one compassionately be there for dying patients. The transforming experience of death as essential to one's self as human being, recognizing that one is living a story with death necessarily a part, promises to overcome the tendency to deny the existential meaning of death for dying patients. This tendency manifests itself through dishonesty about medicine's limitations in creating prognoses, and specifically survival estimates, as well as in holding only a curative and not palliative goal of treatment. This tendency will be replaced by honest and compassionate actions with those in the process of dying. Representing this change is a focus on the patient as person, living a certain lifestyle, and defining himself by significant events and relationships in the past, present, and future. Death and dying become meaningful through incorporation into the story and style that is the patient. This meaning that is facilitated by caregivers and created by patients is central to achieving a "good death." PMID- 12911073 TI - A syllabus for fellowship education in palliative medicine. AB - Recent years have seen significant growth in palliative medicine training programs and positions. There are plans to pursue palliative medicine specialty status with the American Board of Medical Specialties and accreditation of fellowship programs with the American College of Graduate Medical Education. A work group of program directors, supported initially by the Cleveland Clinic and then by the American Board of Hospice and Palliative Medicine, has recently published standards for fellowship training. Despite this, fundamental questions remain about defining the field and delineating the knowledge and skills expected following completion of specialty training. In this article, we describe the first fellowship program in palliative medicine (PMP) in the United States, developed and supported by the Cleveland Clinic Foundation. The program has been implemented as part of the Harry R. Horvitz Center for Palliative Medicine, founded in 1987 as the first comprehensive integrated US program in this field. This training program, in existence since 1989, features a traditional rotational structure with an inpatient primary care service, inpatient consult services, and an outpatient consult/hospice service. This article outlines the syllabus developed for this fellowship, given what we believe to be the essential knowledge base for the field of palliative medicine. PMID- 12911074 TI - Hospice care in prison: general principles and outcomes. AB - In recent years, tougher sentencing laws have resulted in larger numbers of elder prison inmates and, consequently, more deaths occurring in prisons. In this context, the introduction of prison hospice programs takes on great significance. The purpose of this study is to identify the principle components and outcomes of prison hospice programs based on data gathered from semistructured telephone interviews with prison hospice providers in state and federal correctional institutions and from other sources. The results suggest that there is a growing effort to provide palliative care to dying inmates across the country and that all of the existing programs share common elements and similar structures. Major outcomes of prison hospice programs include cost-effectiveness, enhanced correction, and comfort care. PMID- 12911076 TI - Medication-focused approach to total pain: poor symptom control, polypharmacy, and adverse reactions. AB - Neuropathic pain, known to have poor opioid response, can be difficult to control. Although several classes of adjuvant medications are believed to be of benefit in managing neuropathic pain, they have potential side effects that occasionally outweigh their benefits. The psychospiritual suffering of patients with advanced cancer may heighten the distress associated with physical symptoms. If undiagnosed, this may lead to increases in dose and the number of medications administered in the hope of better symptom control. This case report describes the successful interdisciplinary management of an advanced cancer patient whose multiple drug therapy had added to rather than alleviated his distress by causing more side effects than symptom relief. PMID- 12911075 TI - Meloxicam and selective COX-2 inhibitors in the management of pain in the palliative care population. AB - This paper discusses the treatment of pain in the palliative care patient, specifically the use of meloxicam and recent advances in agents with cyclooxygenase-2 (COX-2) selectivity. Meloxicam is a nonsteroidal anti inflammatory drug (NSAID) that preferentially inhibits COX-2 more than cyclooxygenase-1 (COX-1), especially at low doses, thereby offering advantages over traditional nonselective NSAIDs. New COX-2 selective agents are discussed, including valdecoxib, parecoxib, etoricoxib, and COX-189. PMID- 12911077 TI - "Good-bye, my friend". PMID- 12911078 TI - Occupational therapist home evaluations: inequalities, but doing the best we can? AB - OBJECTIVE: The purpose of this study was (a) to describe the occupational therapy recommendations provided to patients discharged to inner city homes, and (b) to examine the relationship between patient health insurance and the number and type of occupational therapist recommendations for equipment and home modifications. METHOD: An archival review was conducted of all referrals to the home evaluation program (n = 755) at a large urban rehabilitation hospital between January 1, 1994, and December 31, 1998. Additional patient demographic data and Functional Independence Measure (FIM) data were obtained in electronic form from the hospital information database. RESULTS: Analysis of results showed that while the pattern of equipment and modification recommendations varied little, publicly insured patients received fewer home modification recommendations compared to privately insured patients (t = 3.7; p < .0005), and were discharged from rehabilitation with significantly lower functional independence (MANOVA F = 3.9; p = .05). CONCLUSION: Results alert occupational therapists to the relationship between health insurance and treatment recommendations and point to patient advocacy and health policy as potential pathways to desired a achieve social change. PMID- 12911079 TI - The development of an assessment to evaluate behavioral factors associated with falling. AB - The purpose of this study was to report the development of the Falls Behavioral (FaB) Scale for Older People, an assessment designed to evaluate behavioral factors that could potentially protect against falling. Instrument development included content analysis, expert review, and factor analysis. Ten behavioral dimensions were identified including Cognitive Adaptations, Protective Mobility, Avoidance, Awareness, Pace, Practical Strategies, Displacing Activities, Being Observant, Changes in Level, and Getting to the Phone. The final 30-item scale had a Content Validity Index of 0.93. Test-retest reliability was ICC = 0.94 (p < .01). Construct validity was established by showing that, as expected, scale scores were positively associated with increasing age (rs = 0.46, p < .01) and negatively associated with greater physical mobility (rs = -0.68, p < .01). People who had fallen utilized safer behaviors than those who had not reported a fall (p < .05) providing a benchmark for using the scale in future studies. The FaB is an easily completed, reliable, and valid tool for determining the presence or absence of protective behaviors. It has potential to assist in goal setting for falls prevention and to evaluate behavioral outcomes of fall prevention programs. PMID- 12911080 TI - Test-retest reliability and convergent validity of the Fatigue Impact Scale for persons with multiple sclerosis. AB - OBJECTIVE: The test-retest reliability and the convergent validity of the Fatigue Impact Scale (FIS) were evaluated using secondary data from 54 persons with multiple sclerosis (MS). METHODS: This reliability and validity study used FIS data from before and after two control periods to evaluate test-retest reliability. Convergent validity of the FIS with the Fatigue Severity Scale and with subscales of the SF-36 Health Survey was evaluated using data collected before the first control period. RESULTS: No significant differences between before and after FIS measurements and intraclass correlation coefficients ranging from .68 to .85 indicate that the FIS has good test-retest reliability except for the physical subscale. The expected moderate correlations between the FIS and several subscales of the SF-36 support its convergent validity. In contrast, the unexpected low correlation between the FIS and Fatigue Severity Scale does not support convergent validity. CONCLUSION: The FIS has adequate reliability and validity and is recommended to evaluate the effectiveness of fatigue management interventions such as energy conservation education for persons with MS. PMID- 12911081 TI - Environmental effects on the assessment of people with dementia: a pilot study. AB - OBJECTIVE: The purpose of this pilot study was to use a standardized assessment of independent living skills to explore the effects of environment on functional performance of individuals with dementia. METHODS: Twelve participants (6 males, 6 females), diagnosed with dementia, were given the Structured Assessment of Independent Living Skills (SAILS), a standardized assessment of functional motor, cognitive, instrumental, and social performance. Participants were assessed in their homes, in an adult day-services facility they regularly attended, and in an occupational therapy clinic. RESULTS: Data were analyzed using repeated measures analysis of variance (ANOVA). There was no evidence of a learning effect from repeated assessments. Participants' performances did not differ among the home, clinic, and adult day-services settings on the total SAILS score (F = 1.22; df = 2,20; p = 0.3176), nor on three of its subscales: cognitive score (F = 0.80; df = 2,20; p = 0.4648), instrumental activities (F = 1.37; df = 2,20; p = 0.2777), and social interaction (F = 0.34; df = 2,20; p = 0.7147). However, participants' performance on the SAILS motor score was significantly higher in the home than in the clinic (t = 2.925, df = 11, p = 0.0138). CONCLUSION. Participants' motor performance was significantly better at home than in an unfamiliar environment. Effects of environment on motor performance, and absence of effects on cognitive, instrumental, and social performances, can be explained through ecological theory. These results suggest that the ability to adapt movement to an unfamiliar environment may decline with the onset and progression of dementia. PMID- 12911082 TI - Impact of the social environment on occupational experience and performance among persons with unilateral neglect. AB - OBJECTIVE: The purpose of this study was to gain an understanding of social environmental influences on occupational experience and performance over the course of rehabilitation of four persons with unilateral neglect. METHODS: The Phenomenological Psychological Method (Karlsson, 1993) was used to gather and analyze qualitative data on four women over a 4-month period. Four women were interviewed five to seven times each and observed during their rehabilitation process. Analysis focused on how these persons with unilateral neglect experienced and responded to their interactions with others in their everyday lives. FINDINGS: As perceived by the study participants, rehabilitation professionals and relatives demonstrated a sequence of strategies for dealing with and helping the women perform despite their unilateral neglect. The sequence of strategies reflects the changing experience of neglect over time. The strategies used by professionals and family members during the rehabilitation process enabled these women to reclaim and occupy the neglected half of the world. Moreover, the women with neglect learned to incorporate others as extensions of their bodies in order to compensate for their ongoing inability to directly experience the left half of the world. CONCLUSION: The results suggest that it is important for others (e.g., therapists, nurses, or relatives) to understand the client's changing experience of neglect and adjust their support and collaboration accordingly. Occupational therapy intervention for persons with neglect could potentially become more effective by systematically incorporating the kinds of strategies identified in this study. Moreover, therapists could consult with others to increase their understanding and effectiveness in interacting with persons who have unilateral neglect. PMID- 12911083 TI - Clara: a phenomenology of disability. AB - A phenomenological approach was used to describe the lived experience of disability of a woman who sustained a head injury 21 years ago. Data were collected through 11 face-to-face interviews and 72 e-mail messages over the period of 11 weeks. An iterative, fluid process of questioning, information giving, analysis, and verification was characteristic of the whole study, alternating between analysis of the particular and analysis of the general. Through a process of phenomenological reduction, each interview transcript and e mail message was divided into meaning units that denoted shifts of meaning. Similar meaning units within a transcript or message were then grouped and summarized in a phrase or word. As new meaning units emerged, additional interviews and e-mail exchanges took place in order to obtain an exhaustive description of the experience of disability. All meaning units obtained from this iterative process finally were "horizonalized" into meaning clusters of nonrepetitive themes. The themes of nostalgia, abandonment, and hope emerged through this collaborative process. A shift in view of life was noted in the participant as the reflective process of this study unfolded, suggesting a phenomenological collaboration between patient and therapist may engender a more genuine connection in which personal meaning is authentically the cornerstone of occupation-centered treatment. PMID- 12911084 TI - The significance of being occupied: the social construction of childhood occupations. AB - The purpose of this paper is to explore theoretical and developmental foundations for interpreting childrens hood engagement with activity in the typically social worlds of childhood. Drawing upon longitudinal ethnographic children data, I argue for the need to reframe the study of childhood occupation to the study of "socially occupied beings" as a means of enhancing our understanding of children's experiences. The focus is on childhood experiences that are socially constructed through adult and child co-created action sequences. The unit of analysis is constructed around a child or children, their adult partners in action, the social world of engagement, and proposed the cultural context. The interpretive focus is on acts and actors, acting in a socially constructed world. Two pation, microethnographic examples are provided; the first relates to an observation of a mother and her children playing in a hospital corridor and the second to an occupational therapist and child engaged in jointly constructed activity within a therapy session. These segments illustrate pragmatic and conceptual understandings of the interconnectedness of social relatedness, intersubjectivity, social action, and engagement. Further development of theoretical and research models is needed to capture the essence of children as socially occupied beings, doing something with someone else that matters. PMID- 12911085 TI - Occupational therapy in transitioning adolescents to post-secondary activities. AB - OBJECTIVE: The purpose of this study was to examine the perceptions of special education directors on the current role of occupational therapy in high school transition programs for adolescent students with disabilities. Additionally, barriers to providing occupational therapy services and perceptions about new occupational therapy services were examined. METHOD: A mailed questionnaire was administered to all special education directors in a rural state in the United States. One hundred and four (57.5%) responses were received. Descriptive statistics were generated with an emphasis on percentages to examine current occupational therapy services in high school transition programs and barriers to service delivery. RESULTS: In this study, occupational therapists provided less than one fifth of transition services in high schools for students with disabilities. They provided more assistive technology consults (30.3%), task or environmental modification (25.8%), and Individualized Education Plan (IEP) and Individualized Transition Plan (ITP) planning (20%) than other providers. Barriers to occupational therapy use included funding, lack of inter-agency planning, and lack of parent participation. About 35% of special education directors suggested that additional occupational therapy services were needed for adolescents with cognitive disabilities and for job performance and related work skills programming. CONCLUSION: Occupational therapists in this study provided ancillary services to high school students with disabilities with greater emphasis on technology, task or environmental modification, and IEP or ITP planning, as perceived by special evaluation directors. PMID- 12911086 TI - Children with disturbances in sensory processing: a pilot study examining the role of the parasympathetic nervous system. AB - This study was a preliminary investigation of parasympathetic nervous system (PNS) functioning in children with disturbances in sensory processing. The specific aims of this study were to (1) provide preliminary data about group differences in parasympathetic functions, as measured by the vagal tone index, between children with disturbances in sensory processing and those without; (2) determine effect size and power needed for future studies; and (3) to lay the foundation for further examination of the relations of parasympathetic functioning and functional behavior in children with disturbances in sensory processing. Participants were 15 children, nine with disturbances in sensory processing and six typically developing children. Heart period data were continuously collected for a 2-minute baseline and during administration of the 15-minute Sensory Challenge Protocol, a unique laboratory protocol designed to measure sensory reactivity (Miller, Reisman, McIntosh, & Simon, 2001). Groups were compared on vagal tone index, heart period, and heart rate using two-tailed, independent sample t tests. Children with disturbances in sensory processing had significantly lower vagal tone than the typically developing sample (t(13) = 2.4, p = .05). Statistical power analysis indicated that, for future studies, a sample size of 20 in each group would yield adequate statistical power. Although the number of subjects in this pilot study is small, the results from this study support further investigations of parasympathetic functions and functional behavior in children with disturbances in sensory processing. PMID- 12911088 TI - Relationship between visual-motor integration and handwriting skills of children in kindergarten: a modified replication study. AB - OBJECTIVE: The purpose of this study was to examine the relationship of performance on the Developmental Test of Visual-Motor Integration (VMI; Beery, 1997) to handwriting legibility in children attending kindergarten. The relationship of using lined versus unlined paper on letter legibility, based on a modified version of the Scale of Children's Readiness in PrinTing (Modified SCRIPT; Weil & Cunningham Amundson, 1994) was also investigated. METHOD: Fifty four typically developing kindergarten students were administered the VMI; 30 students completed the Modified SCRIPT with unlined paper, 24 students completed the Modified SCRIPT with lined paper. Students were assessed in the first quarter of the kindergarten school year and scores were analyzed using correlational and nonparametric statistical measures. RESULTS: Strong positive relationships were found between VMI assessment scores and student's ability to legibly copy letterforms. Students who could copy the first nine forms on the VMI performed significantly better than students who could not correctly copy the first nine VMI forms on both versions of the Modified SCRIPT. CONCLUSION: Visual-motor integration skills were shown to be related to the ability to copy letters legibly. These findings support the research of Weil and Cunningham Amundson. Findings from this study also support the conclusion that there is no significant difference in letter writing legibility between students who use paper with or without lines. PMID- 12911087 TI - Practitioners' reading patterns, attitudes, and use of research reported in occupational therapy journals. AB - Occupational therapists are encouraged to use research-based evidence to guide practice. In this study, we investigated whether members of the American Occupational Therapy Association read their flagship journal, the American Journal of Occupational Therapy (AJOT, or other scholarly journals. Therapists' attitudes about research, their reading patterns, and their use of research in clinical practice were also explored. A proportional, random sample of 626 therapists from five states was mailed a questionnaire; 52% were returned. Of the 328 respondents, 85% reported reading AJOT: For those who did not read AJOT, barriers cited included time constraints, difficulty interpreting results, lack of clinical information, and too much scientific information. Attitudes about research generally were positive, although ratings regarding the usefulness of research to inform clinical practices were less favorable. Attitudes about and use of research ratings were not strongly related to practice settings, educational degree level, years of experience, or state of residence. Results suggest education and clinical practice changes may be necessary to support therapists' use of empirical evidence in practice contexts. PMID- 12911089 TI - Mothers' and fathers' interactions with children with motor delays. AB - OBJECTIVE: In early intervention programs, parents are often asked to teach their child new skills. As fathers are increasingly involved in intervention, clinicians need more information on fathers' unique interactive style. This pilot study compared mothers' and fathers' parent-child interactions during a teaching episode to identify similarities and differences in order to better understand parents' strengths. METHODS: The Nursing Child Assessment Teaching Scale was used to observe 10 mothers and 10 fathers interacting with their 10- to 28-month-old children in their homes. The children were receiving early intervention for a motor delay. The Caregiver Scores (parent's contribution to the interaction) of mothers and others were compared using paired t tests. RESULTS: Mothers had more optimal interactions as indicated by significantly higher Caregiver scores than fathers, t (9) = 3.83, p = .004. The subscales with statistically significant differences were Caregiver Contingency and Cognitive Growth Fostering. Children's scores when they interacted with their mothers or fathers did not differ. CONCLUSION: When observing fathers teaching their child new skills, therapists should remember that fathers of children with motor delays (and typically developing children) may use a more task-oriented communication style with less consideration of the child's actions than do mothers. PMID- 12911090 TI - Critical analysis of a public policy: an occupational therapist's experience with the Patient Bill of Rights. PMID- 12911091 TI - The process of evidence-based clinical decision making in occupational therapy. PMID- 12911092 TI - Double-phase Tc-99m MIBI scintigraphy as a therapeutic predictor in patients with non-Hodgkin's lymphoma. AB - PURPOSE: Tc-99m MIBI is a promising agent for the functional characterization of P-glycoprotein expression and the prediction of therapeutic outcome in human tumors. The purpose of this study was to determine whether tumor clearance of Tc 99m MIBI may have a predictive value in the clinical outcome in patients with non Hodgkin's lymphoma. METHODS: Seventeen patients with histologically proved non Hodgkin's lymphoma underwent Tc-99m MIBI scintigraphy before starting chemotherapy. Early and late images were obtained after the injection of 740 MBq (20 mCi) Tc-99m MIBI. Tumor-to-normal soft tissue ratios were measured on early and late images, and the percentage retention (%R) was calculated. These findings were correlated with other prognostic factors. RESULTS: Of all 40 tumor lesions, 22 (55%) showed Tc-99m MIBI tumor uptake, whereas the other 18 (45%) lesions had no uptake. According to the location, the diagnostic accuracy of Tc-99m MIBI scans was 66.7% (12 of 18) for lesions of the head and neck, 66.7% (8 of 12) for those of the chest, and 20% (2 of 10) for those of subdiaphragmatic regions (P = 0.037). The mean %R of complete responders was 91.8 +/- 14.8% and for partial responders or nonresponders it was 65.4 +/- 17.7% (P = 0.014). The %R of Tc-99m MIBI showed a significant negative correlation with the serum lactate dehydrogenase level (r = -0.757), the score of the international prognostic index (r = -0.643), and the Ann Arbor stage (r = -0.733). CONCLUSION: The slow tumor clearance of Tc-99m MIBI can predict a good response to chemotherapy in patients with non-Hodgkin's lymphoma and differentiate responding and nonresponding tumors early in the course of chemotherapy. PMID- 12911093 TI - Postchemotherapeutic reversibility of hypertrophic osteoarthropathy in a patient with bronchogenic adenocarcinoma. AB - A 54-year-old man with bronchogenic adenocarcinoma had features of the hypertrophic osteoarthropathy syndrome. The syndrome is treated by surgical resection of the primary tumor, with rapid remission of the symptoms within 24 hours after surgery. In the case reported here, the tumor (T3N3M0) could not be resected. Clinical and scintigraphic regression of the hypertrophic osteoarthropathy was evident after chemotherapy. Bone scintigraphy is valuable when determining a patient's response to treatment for this paraneoplastic syndrome. PMID- 12911094 TI - Lung, gastric, and soft tissue uptake of Tc-99m MDP and Ga-67 citrate associated with hypercalcemia. AB - Metastatic calcifications are associated with chronic renal failure, hyperparathyroidism, metastatic neoplasms, hypervitaminosis D, and hypercalcemia of other origins. Bone scanning agents accumulate within these extraskeletal metastatic calcifications. The authors describe two patients with hypercalcemia associated with Tc-99m MDP uptake in the lungs, stomach, and soft tissues. Ga-67 scintigraphy was also performed and showed increased uptake in the same locations as those of Tc-99m MDP, suggesting the existence of an inflammatory process. Despite adequate treatment, only partial resolution of extraskeletal uptake was observed. PMID- 12911095 TI - False-positive ventilation-perfusion scan in a patient with a transplanted lung. AB - A false-positive ventilation perfusion (V/Q) scan in a patient with a transplanted lung and secondary pulmonary artery stenosis is reported. The patient was examined for tachypnea and shortness of breath after single lung transplantation. A V/Q scan revealed multiple perfusion defects in the transplanted lung, with no corresponding ventilation defects. A pulmonary angiogram revealed pulmonary artery stenosis and no pulmonary emboli. The authors describe a false-positive V/Q scan in a patient after lung transplantation. PMID- 12911096 TI - Prevalence of silent gastroesophageal reflux in association with recurrent lower respiratory tract infections. AB - PURPOSE: Gastroesophageal reflux (GER) has long been associated with pulmonary symptoms and diseases. This retrospective analysis was undertaken to determine the prevalence of GER detected by gastroesophageal scintigraphy in children with recurrent lower respiratory tract infections but none of the typical gastroesophageal symptoms of GER. MATERIALS AND METHODS: A total of 312 children (mean age, 2.36 years; age range, 4 months to 11 years) were evaluated. All studies were performed using Tc-99m sulfur colloid (3.7 to 7.4 MBq; 100 to 200 microCi). RESULTS: Of 312 children, 108 (34.6%) had GER on gastroesophageal scintigraphy. The children were separated into four groups, depending on their age. Group 1 (119 children, younger than 1 year), group 2 (131 children, ages 1 to 3 years), group 3 (36 children, ages 3 to 6 years), and group 4 (26 children, ages 6 to 12 years) showed silent GER with prevalences of 38.7%, 37.4%, 22.2%, and 19.2%, respectively. Because physiologic GER is common in children younger than 18 months, we also analyzed the prevalence of GER in children younger than 18 months and older than 18 months of age. The prevalences of GER in these groups were 41.7% and 24.6%, respectively. CONCLUSIONS: A significant number of children with recurrent lower respiratory tract infection have scintigraphic evidence of GER. This high prevalence, even in children older than 18 months, cannot be ignored. Prospective studies including many patients are needed to establish the exact role of GER in these patients. PMID- 12911098 TI - Current readings in nuclear medicine. PMID- 12911097 TI - Evaluation of Tc-99m-labeled glycoprotein IIb/IIIa receptor antagonist DMP444 SPECT in patients with infective endocarditis. AB - PURPOSE: Infective endocarditis (IE) is characterized by aggregation of activated platelets, fibrin, and bacteria. DMP444, a high-affinity glycoprotein IIb/IIIa receptor antagonist, binds to the fibrinogen-binding domain of activated platelets, depicting a key feature of IE. Tc-99m DMP444 scintigraphy was studied in a group of patients with possible IE. METHODS: Tc-99m DMP444 (600 MBq; 16 mCi) planar and SPECT images of the heart were recorded in patients with possible IE for as long as 6 hours after injection. Results were compared to echocardiography and the Duke classification. RESULTS: Sixteen patients (age range, 37 to 78 years) participated. DMP444 imaging was positive on SPECT in five patients, and all had definite endocarditis (affecting both prosthetic and native valves). Eleven patients were DMP444 negative, seven with no proof of IE. The remaining four patients were classified as having IE, but three had been receiving adequate intravenous antibiotic regimens for > or = 2 weeks at the time of scintigraphy and one had Q-fever endocarditis. CONCLUSIONS: DMP444 SPECT allows in vivo visualization of IE if it is performed within 1 to 2 weeks after the start of antibiotic treatment. Given the high affinity of DMP444 for activated platelets, the results indicate the involvement of activated platelets in early IE. PMID- 12911099 TI - Semantic and cross-case identity priming in patients with Parkinson's disease. AB - Semantic and cross-case identity priming were investigated in nondemented patients with Parkinson's disease (PD) and controls using the Lexical Decision Task. Three conditions were administered that consisted of the presentation of prime and target word pairs. In the semantic priming condition the word pairs were semantically related (e.g., table-CHAIR), in the cross-case identity priming condition the word pairs consisted of the same word (e.g., noise-NOISE), and in the unrelated condition the word pairs were not related semantically (e.g., guns DEEP). A fourth condition was also administered that consisted of the presentation of a prime word and a pronounceable nonword target (e.g., starved FORVE). Participants were asked to indicate whether the target was a real word or a nonword. The prime and target were separated by either a short or long (250 ms or 1000 ms) stimulus onset asynchrony (SOA). Results indicated that PD patients displayed normal semantic priming (i.e., faster responding to the target in the semantic condition as compared to the unrelated condition) at both the short and long SOA. Similarly, PD patients displayed normal cross-case identity priming (i.e., faster responding to the target in the identity condition relative to the unrelated condition) at the long SOA. At the short SOA, however, PD patients displayed hyper identity priming relative to controls (134 ms vs. 50 ms). These results suggest that semantic processes are normal in nondemented PD patients but that the processes involved in accessing lexical information may be overly activated in these patients. PMID- 12911100 TI - Speech lateralization and attention/executive functions in a Finnish family with specific language impairment (SLI). AB - Hemisphere lateralization for speech perception was investigated in a Finnish family with specific language impairment. We used dichotic presentations of consonant-vowel (CV) syllables, consisting of the six stop-consonants paired with the vowel /a/, under three different attentional instructions. The dichotic listening technique means that two different speech stimuli are presented simultaneously, one in each ear. Left hemisphere dominance for speech perception is assessed from a preference for the right ear stimulus. Response accuracy was compared in five members, over three generations, of a family with SLI with 5 healthy control subjects. The dichotic listening task was performed under three different instructions, to attend only to the right ear stimulus, to attend only to the left ear stimulus, or with no instruction about attention. The subjects indicated orally which of the six CV-syllables they heard on each trial. There were 36 dichotic presentations of the CV-syllables for each instruction. The CV syllables were played from a CD with digitized stimuli. The results showed no difference between the groups during the no instruction condition, indicating normal speech lateralization in the SLI group. However, the SLI group was deficient in modifying the ear advantage through focused attention to the left ear, thus indicating an attentional/executive deficit in addition to a deficit in left hemisphere lateralization. It is concluded that individuals with specific language impairment may have a cognitive deficit related to attention in addition to a language processing deficit related to left hemisphere function. PMID- 12911101 TI - Using the WMS-III faces subtest to detect malingered memory impairment. AB - The current study evaluated the utility of the WMS-III Faces I subtest (Faces) for the assessment of malingering. Thirty nonlitigating traumatic brain injury patients and 30 control participants were administered Faces under standard administration and instructed malingering conditions. Although the two groups obtained similar scores when taking the test under standard instructions, both groups produced significantly lower performances when instructed to malinger, indicating that Faces is sensitive to malingering, but less sensitive to traumatic brain injury. The total raw score provided stronger classification accuracy than an empirically weighted combination of the five easiest items (i.e., floor effect items). A raw score cutoff of 31 yielded the maximum classification accuracy with 93.3% sensitivity and 80.0% specificity. PMID- 12911102 TI - Memory performance, but not information processing speed, may be reduced during early pregnancy. AB - Several studies have investigated aspects of cognitive functioning during late pregnancy or in the period around delivery. The present paper describes a controlled study of neurocognitive functioning in an early phase of pregnancy (14 weeks). Seventy-one pregnant women and 57 control subjects matched for age and education were tested with a cognitive test battery. Intentional learning was tested with the Verbal Learning Test, retrieval from semantic memory with the Fluency Test, and speed of information processing with the Concept Shifting Test, the Stroop test, and the Letter Digit Substitution Test. Results show that performance on tests measuring intentional learning and retrieval from semantic memory were lower in the pregnant group during early pregnancy as compared to a closely matched nonpregnant group. In contrast, speed of information processing was not different between the two groups. The differences observed in memory performance were not large and further research is needed to establish their clinical significance. In addition, the results should be interpreted with care, because our study has a cross-sectional design, which has limitations concerning the fact that preexisting performance differences might be possible. Therefore, longitudinal studies are essential to ascertain clear associations between pregnancy and cognitive performance. PMID- 12911103 TI - Differentiating simple versus complex processing speed: influence on new learning and memory performance. AB - The current study was designed to examine how the construct of human information processing speed is conceptualized and measured, while also examining the influence of information processing speed on higher cognitive processes (i.e., learning). A mixed medical sample of 92 subjects participated in this study. Subjects underwent a broad-based neuropsychological evaluation, including measures of verbal and visuospatial new learning, spatial and verbal working memory, simple reaction time, choice reaction time, and information processing speed. Principal components factor analysis with varimax rotation resulted in a three-factor solution, comprised of: (1) simple speed/reaction time, (2) complex information processing and new learning, and (3) working memory. Notably, this factor solution identified 2 distinct forms of processing speed--simple and complex information processing speeds. In contrast to the abundance of literature grouping these two constructs together under one term (i.e., processing speed), these results indicate simple and complex speed to be distinct constructs assessed with different neuropsychological instruments. While the expected relationship between complex information processing capacities and working memory abilities was evident in this study, information processing speed also showed a significant relationship with new learning ability. The implications of this intriguing relationship are discussed. PMID- 12911105 TI - Use of the WCST and the WCST-64 in the assessment of traumatic brain injury. AB - The Wisconsin Card Sorting Test (WCST) has been found to be valid in characterizing cognitive dysfunction in a variety of neurological disorders including traumatic brain injury (TBI). However, the test has been criticized as being too lengthy and frustrating for severely impaired patients. As a result, shortened versions of the WCST have been proposed. The utility of one of these shortened versions, the Wisconsin Card Sorting Test-64 (WCST-64), was examined in 106 persons with TBI. Findings showed strong associations between scores derived from the two tests. WCST scores were predicted from WCST-64 scores with sufficient accuracy for research purposes with adjusted R-squared values ranging from .74 to .87. Using the standard cutpoint of < 40T to indicate impairment or normal performance for perseverative responses from each of the two tests, 91 (86%) of 106 subjects received the same classification showing substantial agreement (Kappa statistic 0.71; 95% CI 0.58-0.84). The WCST and the WCST-64 also performed comparably in predicting functional status at discharge from inpatient rehabilitation using the first score from a principal components analysis as a summary measure (both significant with p = .0002). These findings support theuse of the WCST-64 in early evaluations of persons with moderate and severe TBI. PMID- 12911104 TI - Central coherence and cognitive shifting in relation to social improvement in high-functioning young adults with autism. AB - The objective of this prospective study was to evaluate the possible role of two cognitive styles--weak central coherence and poor cognitive shifting--in predicting social improvement in patients with autistic disorder. Thirty patients, largely similar in age (young adults), intelligence (high-functioning) and living conditions (residential treatment in the same unit) were assessed at two separate time points with a 3-year interval between pretest and posttest. At pretest central coherence, cognitive shifting and several aspects of social functioning--symptom severity, social intelligence and social competence--were measured. At posttest social functioning was reassessed. Unlike central coherence, cognitive shifting was identified as a significant prognostic marker. This differential outcome might be an indication that patients with poor cognitive shifting and patients with weak central coherence have different prognoses with the current, highly structured treatment milieu; it is unknown whether patients with poor cognitive flexibility might benefit more from treatments specifically designed to address this problem. PMID- 12911106 TI - The Snodgrass and Vanderwart set revisited: norms for object manipulability and for pictorial ambiguity of objects, chimeric objects, and nonobjects. AB - In this paper, we propose a standardized set of 480 black-and-white line drawings, half meaningful and half meaningless. Meaningful pictures represent a common object, and were selected from the Snodgrass and Vanderwart set (1980). Meaningless pictures include 120 chimeric objects (made up of two halves of real objects) and 120 nonobjects, that were constructed from the meaningful pictures while controlling for visual complexity. We report the results of two experiments designed to standardize the revisited Snodgrass and Vanderwart set along two important dimensions for picture processing: object manipulability (Experiment 1) and pictorial ambiguity (Experiment 2). The relevance of these dimensions is discussed. Experiment 1 permit us to sort objects into four manipulability categories (i.e., the ease and distinctiveness with which use of the object can be mimed) and to propose a manipulability index. This experiment provides additional evidence for a partial overlap in the dichotomy between man-made objects and living things, on the one hand, and manipulable and unmanipulable objects, on the other hand. In Experiment 2, a pictorial ambiguity index was computed for meaningful and meaningless pictures. The results of this experiment point the distinction between chimeric objects and nonobjects showing that chimeric objects are more complex to process than nonobjects and objects. This standardized set of pictures provides a database and an hopefully useful tool for research in cognitive neuroscience. PMID- 12911107 TI - Rhythmic oscillations in the performance of a sustained attention task. AB - Attempts to sustain a narrow focus of attention over a long period of time are effortful and are punctuated by lapses. Most studies of sustained attention performance obscure the presence and pattern oflapses by reporting measures that are summed across the entire period of an individual's performance, or that are average scores for blocks of trials across many participants. In the present study we attempted to explore fluctuation in the attention of individual participants over the course of a vigilance task and to quantify its periodicity, if any exists. Normal university students listened to letters of the alphabet, arranged randomly and presented at a rate of 2 per second for 20 min. They were instructed to press a hand-held button when they detected a target two consecutive identical letters). Continuous estimates of performance accuracy (correctly detected targets) at regularly spaced time intervals were created for each participant using a moving time window. The resulting functions were analyzed in order to detect and quantify periodicity using a Fast Fourier Transform (FF). The most often observed rhythms for those participants with adequate FFT power congregated at 1-2 min, 4-7 min and greater than 10 min. Performance functions from 36 of the 40 subjects displayed at least two of these frequencies. Other studies have identified cycles in performance during similar vigilance challenges, but without particular rhythms or with no particular shared frequencies amongst participants. The possible sources of these fluctuations and the differences in the findings of these studies and the present study are discussed. PMID- 12911108 TI - The 50 and 100-item short forms of the Paced Auditory Serial Addition Task (PASAT): demographically corrected norms and comparisons with the full PASAT in normal and clinical samples. AB - While the standard 200-item version of the Paced Auditory Serial Addition Task (PASAT) is a sensitive neuropsychological instrument, it can be quite aversive to some patients due to its length and progressively increasing difficulty. We present demographically-corrected norms for 50 and 100-item short-form versions in a sample of 560 neurologically normal adults. Age, education, and ethnicity (but not gender) were found to be significant predictors of performance. In a clinical sample of 786 HIV-infected adults, diagnostic accuracy of the 50, 100, and 200-item versions was essentially equivalent (using clinical ratings of a comprehensive neuropsychological battery as the gold standard, overall classification rates of the three PASAT versions were 71%, 74%, and 73%, respectively), with better specificity (89-92%) than sensitivity (46-53%). The 50 item version showed moderate ceiling effects, but the 100-item test did not. In a mixed clinical sample of 40 subjects, the 50-item version was administered more than twice as fast as the 200-item version, and was tolerated better (discomfort rating of 4.0 vs. 5.9 on a 10-point scale, p < .05). We conclude that in many cases the PASAT-50 and PASAT-100 provide equivalent diagnostic accuracy with a significant reduction in administration time and patient discomfort. PMID- 12911109 TI - Mitochondrial distribution and glycogen dynamics suggest diffusion constraints in muscle fibers of the blue crab, Callinectes sapidus. AB - The scaling of mitochondrial distribution, citrate synthase activity, and post contractile glycogen recovery was examined in muscle fibers of the blue crab, Callinectes sapidus. The fast-twitch muscle fibers of C. sapidus can reach extremely large dimensions, which may impose constraints on aerobic metabolic processes. However, muscle cells from small crabs are not giant, meaning that during development muscle fibers cross and greatly exceed the surface area to volume (SAV) and diffusion threshold that is adhered to by the cells of most organisms. Cell diameters in the smallest size class were approximately 100 microm, while the largest size class had cell diameters in excess of 500 microm. In the smallest cells, the fractional area of subsarcolemmal and intermyofibrillar mitochondria was similar. However, in the largest cells, mitochondria were almost exclusively subsarcolemmal. Total fractional area of mitochondria was highest in the largest cells due to a proliferation of subsarcolemmal mitochondria. In contrast, citrate synthase activity decreased as cell size increased. Following burst contractile activity, glycogen concentrations decreased significantly and remained depressed for several hours in muscle comprised of giant cells, consistent with previous findings that anaerobic glycogenolysis fuels certain components of post-contractile recovery. However, in muscle composed of the smallest muscle cells, glycogen levels did not decrease significantly following burst activity. While normal scaling of aerobic metabolism would predict a slower aerobic recovery in larger animals, the present results suggest that cellular organization, SAV, and intracellular diffusion distances also impose constraints on aerobic processes in C. sapidus. PMID- 12911110 TI - Vascular anatomy of the gills in a high energy demand teleost, the skipjack tuna (Katsuwonus pelamis). AB - Tunas (family: Scombridae, Tribe: Thunnini) exhibit anatomical, physiological, and biochemical adaptations that dramatically increase the ability of their cardiorespiratory systems to transfer oxygen from the water to the tissues. In the present study the vascular anatomy of the skipjack tuna, Katsuwonus pelamis, gill was examined by light and scanning electron microscopic analysis of methyl methacrylate vascular corrosion replicas prepared under physiological pressure. The gill filament contains three distinct blood pathways, respiratory, interlamellar, and nutrient. The respiratory, or arterio-arterial (AA) pathway, is the site of gas exchange and consists of the afferent and efferent filamental arteries (AFA and EFA) and arterioles (ALA and ELA) and the lamellae. Each ALA in the basal filament supplies ten or more lamellae and they anastomose with their neighbor to form a continuous vascular arcade. Four modifications in the lamellar circulation appear to enhance gas exchange efficiency. 1) The ALA deliver blood directly to the outer margin of the lamellae where unstirred boundary layer effects are predicted to be minimal and water PO2 highest. 2) Pillar cells are closely aligned along the outer boundary of the inlet side and the inner boundary of the outlet side of the lamellae to form multiple distributing and receiving blood channels. 3) Elsewhere in the lamella, pillar cells are aligned to form diagonal channels that direct blood from the outer to the inner lamellar margins, thereby reducing vascular resistance. 4) The lamellar sinusoid is especially widened near the efferent end to augment oxygen saturation of blood flowing through the inner margin. These adaptations, plus the presence of a bow-shaped interlamellar septum, and a thinned filament core appear to decrease gill vascular resistance and maximize gas-exchange efficiency. The interlamellar (IL) and nutrient systems originate from post-lamellar vessels and are arterio-venous (AV) pathways. IL vessels form an extensive ladder-like lattice on both sides of the filamental cartilage and are supplied in part by narrow-bore vessels from the medial wall of the EFA. Their function is unknown. Nutrient vessels are formed from the confluence of a myriad of tortuous, narrow-bore vessels arising from the basal region of the EFA and from efferent branchial arteries. They re-enter the filament and eventually drain into the IL system or filamental veins. As these AV pathways are retained despite considerable reduction in filamental tissue, it is evident that they are integral components of other non-respiratory homeostatic activities of the gill. PMID- 12911111 TI - Differential physiological expression of the invertebrate 2Na+/1H+ antiporter in single epithelial cell type suspensions of lobster hepatopancreas. AB - Lobster (Homarus americanus) hepatopancreas is a complex, heterogeneous tissue composed of four epithelial cell types that individually contribute to the overall functional properties of digestion, absorption, secretion, and detoxification. Previous studies, using purified hepatopancreatic brush border membrane vesicles, have described the properties of an electrogenic, 2Na+/1H+ antiporter in this tissue that regulates the absorption and secretion of these cations. These studies were not able to localize this cation exchange phenomenon to specific epithelial cell types. In the present study, sodium/proton exchange by purified, single cell, suspensions of lobster (Homarus americanus) hepatopancreatic epithelium was investigated using a centrifugal elutriation method to cleanly separate the four individual cell types for subsequent physiological characterization. Results indicate that all four hepatopancreatic epithelial cell types possessed the 2Na+/1H+ antiporter as a result of its unique sigmoidal influx properties. Hill Coefficients, measures of transport sigmodicity obtained from kinetic analyses of 22Na+ influx by single cell type suspensions, varied from 1.56 +/- 0.30 (R-cell suspensions) to 2.79 +/- 0.41 (F-cell suspensions), suggesting that different numbers of sodium ions may be accommodated by each cell type. Both calcium and zinc were competitive inhibitors of 22Na+ influx in E-cells (calcium Ki = 105.1+/-5.2 microM; zinc Ki = 46.2 +/- 7.8 microM), but the extent to which these divalent cations inhibited monovalent cation transport by each cell type varied. It is concluded that different isoforms of the electrogenic 2Na+/1H+ antiporter may be present in each hepatopancreatic cell type and thereby contribute in differing degrees to the cation regulatory functions performed by the overall organ. PMID- 12911112 TI - Characterization of the effects of vasoactive substances on the bulbus arteriosus of the eel, Anguilla rostrata. AB - The fish bulbus arteriosus (BA) smooths cardiac output by expanding during cardiac systole and rebounding during diastole, thereby providing constant perfusion of the gills downstream. Published data have demonstrated innervation of the teleost BA and shown that the tension and compliance of the BA responded to vasoactive agonists, such as epinephrine and acetylcholine, suggesting that the BA was more than a mere "windkessel." To examine vasoactivity in the BA more directly, we measured the responses of isolated tissue rings from the BA of the eel, Anguilla rostrata to a suite of putative vasoactive agonists, which had been shown to affect vascular smooth muscle in a variety of teleosts. The BA of the eel was insensitive to acetylcholine but constricted when endothelin (ET-1) was applied. Nitric oxide, sodium nitroprusside (SNP; NO donor), natriuretic peptides (NP), and prostaglandin E1 (but not the prostacyclin agonist carbaprostacyclin) produced significant dilation in the BA. Since both ET-1 and sarafotoxin S6c produced concentration-dependent constriction, it appears that endothelin receptor B-type (ETB) receptors (and possibly ETA receptors) are present. The dilation produced by SNP was also concentration dependent, as were the dilations produced by porcine C-type natriuretic peptide, eel atrial natriuretic peptide (NP receptor agonists), Sulprostone and Butaprost (PGE receptor agonists). Our data demonstrate that the BA of eel is responsive to a variety of vasoactive agonists, suggesting that the BA is under neurohumoral control. The role of agonist-induced changes in BA tension in fish cardiovascular physiology remains to be determined, as do the specific receptor types involved. PMID- 12911114 TI - Responses to amputation of denervated ambystoma limbs containing aneurogenic limb grafts. AB - The developing neural tubes and associated neural crest cells were removed from stage 30 Ambystoma maculatum embryos to obtain larvae with aneurogenic forelimbs. Forelimbs were allowed to develop to late 3 digit or early 4 digit stages. Limbs amputated through the mid radius-ulna regenerated typically in the aneurogenic condition. Experiments were designed to test whether grafts of aneurogenic limb tissues would rescue denervated host limb stumps into a regeneration response. In Experiment 1, aneurogenic limbs were removed at the body wall and grafted under the dorsal skin of the distal end of amputated forelimbs of control, normally innervated limbs of locally collected Ambystoma maculatum or axolotl (Ambystoma mexicanum) larvae. In Experiment 1, at the time of grafting or 1, 2, 3, 4, 5, 7, or 8 days after grafting, aneurogenic limbs were amputated level with the original host stump. At 7 and 8 days, this amputation included removing the host blastema adjacent to the graft. The host limb was denervated either one day after grafting or on the day of graft amputation. These chimeric limbs only infrequently exhibited delayed blastema formation. Thus, not only did the graft not rescue the host, denervated limb, but the aneurogenic limb tissues themselves could not mount a regeneration response. In Experiment 2, the grafted aneurogenic limb was amputated through its mid-stylopodium at 3, 4, 5, 7, or 8 days after grafting. By 7 and 8 days after grafting, the host limb stump exhibited blastema formation even with the graft extending out from under the dorsal skin. The host limb was denervated at the time of graft amputation. When graft limbs of Experiment 2 were amputated and host limbs were denervated on days 3, 4, or 5, host regeneration did not progress and graft regeneration did not occur. But, when graft limbs were amputated on days 7 or 8 with concomitant denervation of the host limb, regeneration of the host continued and graft regeneration occurred. Thus, regeneration of the graft was correlated with acquisition of nerve-independence by the host limb blastema. In Experiment 3, aneurogenic limbs were grafted with minimal injury to the dorsal skin of neurogenic hosts. When neurogenic host limbs were denervated and the aneurogenic limbs were amputated through the radius/ulna, regeneration of the aneurogenic limb occurred if the neurogenic limb host was not amputated, but did not occur if the neurogenic limb host was amputated. Results of Experiment 3 indicate that the inhibition of aneurogenic graft limb regeneration on a denervated host limb is correlated with substantial injury to the host limb. In Experiment 4, aneurogenic forelimbs were amputated through the mid-radius ulna and pieces of either peripheral nerve, muscle, blood vessel, or cartilage were grafted into the distal limb stump or under the body skin immediately adjacent to the limb at the body wall. In most cases, peripheral nerve inhibited regeneration, blood vessel tissue sometimes inhibited, but other tissues had no effect on regeneration. Taken together, the results suggest: (1) Aneurogenic limb tissues do not produce the neurotrophic factor and do not need it for regeneration, and (2) there is a regeneration inhibiting factor produced by the nerve-dependent limb stump/blastema after denervation that prevents regeneration of aneurogenic limbs. PMID- 12911113 TI - Compensation for hypercapnia by a euryhaline elasmobranch: effect of salinity and roles of gills and kidneys in fresh water. AB - Specimens of the euryhaline elasmobranch, Dasyatis sabina were acclimated to seawater and fresh water, and exposed to normocapnic (air) and hypercapnic (1% CO2 in air) environmental water. Blood pH, PCO2, and [HCO3-], as well as whole animal net-acid excretion, were measured for up to 24 h of hypercapnia. In a separate experimental series, urine was collected from freshwater acclimated stingrays during 8 h of normocapnia and hypercapnia. Stingrays in both salinities at least partially compensated for the respiratory acidosis by accumulating HCO3- in their extracellular spaces. The degree of compensation for blood pH was 88.5% in seawater, but only 31.0% in fresh water after 24 h of hypercapnia. Whole animal net-acid excretion was also greater in seawater than in fresh water, as was the increase in extracellular fluid [HCO3-]. Mean urinary net-acid excretion rates were slightly negative, and never increased above normocapnic control rates during hypercapnia. Since whole-animal net-acid excretion rates increased with blood [HCO3-], and urinary excretion was always negative, the gills were probably the primary organ responsible for compensation from environmental hypercapnia. The faster, and more complete, compensation for hypercapnia in seawater than in fresh water for this euryhaline elasmobranch is consistent with data for euryhaline teleosts, and probably reflects Na+-dependent mechanisms of branchial acid excretion. PMID- 12911115 TI - Melatonin and locomotor activity in the fiddler crab Uca pugilator. AB - The influence of melatonin on locomotor activity levels was measured in the fiddler crab Uca pugilator. First, activity in untreated, laboratory-acclimated crabs was measured over 48 hours in a 12L:12D photoperiod; this study showed a nocturnal increase in activity. In eyestalk-ablated crabs, overall activity was significantly reduced, and no significant activity pattern occurred. Next, crabs were injected with melatonin or saline (controls) at various times during the 12L:12D photoperiod (0900h, 1200h, and twice at 2100h; each trial was separated by 3-4 days) and monitored for 3 hr post-injection. Control crabs had low activity during early photophase, high at mid-photophase, increasing activity during the first scotophase trial, and decreasing activity during the second scotophase trial. Melatonin had no significant influence on activity when injected during the early-photophase activity trough or early-scotophase activity decline, but significantly increased activity when injected during the mid photophase activity peak and early-scotophase activity incline. Next, crabs were injected during an early scotophase activity trough and monitored throughout the twelve-hour scotophase. Melatonin did not increase activity until the mid scotophase activity increase, approximately 6 hours later, showing that the pharmacological dosage persisted in the crabs' systems and had later effects during the incline and peak of activity but not the trough. Eyestalk-ablated crabs were injected with melatonin or saline during early photo- and scotophase. Melatonin significantly increased activity in the photophase but not the scotophase trial, indicating that the responsiveness to melatonin continues following eyestalk removal, but the timing may not match that of intact crabs. Melatonin may be involved in the transmission of environmental timing information from the eyestalks to locomotor centers in U. pugilator. PMID- 12911116 TI - Viewing chelonian reproductive ecology through acoustic windows: cranial and inguinal perspectives. AB - Ultrasonography is a powerful new tool in reproductive physiology and ecology but quantitative evaluations of acoustic window efficacies are necessary to assess methodological accuracy. We evaluated the efficacy of the left inguinal, right inguinal and cranial acoustic windows for detecting ovarian follicles and oviducal eggs in three tortoise species. Incorporating cranial results improved upon inguinal efficacies for detecting follicles in the three species, and for detecting eggs in Chersina angulata, which had small inguinal apertures for its body size. The cranial window provided the best follicle results (highest numbers detected) but the left inguinal window provided the best egg results. This asymmetry was probably due to the primarily posterior position of the oviducts (eggs) and the pliable ovarian sheets allowing follicles to move cranially. The left-right asymmetry may result from intestinal asymmetry (i.e., between the cecum and descending colon) in obscuring ultrasound transmission or displacing follicles and eggs. Seasonal influences upon efficacies varied with species, partially due to species differences in a) reproductive cycles and b) size and number of follicles and eggs. We attributed other efficacy differences among species to differences in body size and shell morphology, especially the inguinal apertures. Correlation analyses revealed strong individual influences upon window efficacy, suggesting that individuals varied in the position of follicles and eggs and the degree of obfuscation by nonreproductive structures. The paucity of correlations between efficacies and body size reflects the complex interactions of body size, shell morphology, reproductive structures and reproductive cycles on efficacies. PMID- 12911118 TI - Cell-based therapy for liver diseases. AB - Although liver transplantation has become standard therapy in the treatment of patients with liver failure, several problems should be considered in the management of these patients. Other approaches have been proposed, in particular cellular-based procedures. Isolated hepatocytes may be used instead of whole organ transplantation or integrated within the bioartificial devices, in order to replace the missing synthetic and metabolic liver functions. Moreover patient's own hepatocytes may be ex vivo genetically modified to provide the function of a mutant gene. However, new cell sources alternative to adult hepatocytes are actually under investigation, on the basis of recent advances in the field of liver repopulation. Xenogenic primary cells, human hepatoma cells, immortalized hepatocytes and stem cells have been testing in several experiments, even if up to now none of them represent a "gold-standard" for cell-based treatment of liver diseases. In the next future, it is possible that different clinical situations will require different therapeutic approaches, that will be finally defined from the concomitant advances in the development of artificial devices and liver cell biology. PMID- 12911117 TI - Discovery of the hepatic canalicular and intestinal cholesterol transporters. New targets for treatment of hypercholesterolemia. AB - Arteriosclerosis and cholesterol cholelithiasis are characterized by abnormal regulation of cholesterol trafficking and solubilization, and subsequent development of the arteriosclerotic plaque in the artery walls and gallstone formation in the gallbladder, respectively. Cholesterol metabolism is controlled by many complex polygenetic - environmental interactions that contribute to the regulation of serum lipoprotein cholesterol levels and biliary cholesterol and bile acids secretion, which constitute the only pathway for sterol elimination from the organism. Much of our understanding of cholesterol metabolism has arisen from studies of the pathways controlling cholesterol synthesis and the uptake and degradation of LDL and HDL lipoproteins. Recently, two new members of the ABC transporter family (ABCG5 and ABCG8 heterodimers) have been discovered in the apical pole of the enterocyte and in the canalicular membrane of hepatocytes. Experiments in genetically engineered mice have demonstrated that ABCG5/G8 represent the physiological canalicular transporter of biliary cholesterol and the intestinal secretory mechanism of absorbed dietary plant sterols. Interestingly, mutation of ABCG5 and or ABCG8 genes in man causes sitosterolemia, a rare genetic disease characterized by massive absorption of plant sterols and premature arteriosclerosis. The potential pharmacological manipulation of biliary cholesterol secretion represents another important therapeutic target to treat hypercholesterolemia, if this manipulation is simultaneously accompanied by measures aimed to avoid gallbladder cholesterol crystallization. The best theoretical drug should decrease serum lipoprotein cholesterol levels, increase biliary cholesterol secretion and fecal elimination and favoring at the same time gallbladder emptying to prevent gallstone formation. PMID- 12911119 TI - Improved control of asthma symptoms with a reduced dose of HFA-BDP extrafine aerosol: an open-label, randomised study. AB - BACKGROUND: Extrafine aerosols may improve asthma symptom control through increased lung deposition of medication to inflamed peripheral airways. METHODS: The effect of switching patients with asthma maintained on up to 2000 microg/day chlorofluorocarbon-beclomethasone dipropionate (CFC-BDP), 1600 microg/day budesonide, 1000 microg/day fluticasone, or 2000 microg/day flunisolide, to a reduced dose of hydrofluoroalkane-134a BDP (HFA-BDP) extrafine aerosol (maximum 800 microg/day) was investigated during an open-label multicentre study. Following a 7-14-day run-in on previous medication, 716 patients were randomised to 24 weeks' treatment with an appropriate reduced dose of HFA-BDP. RESULTS: Morning peak expiratory flow (AM PEF) measurements showed that, after 24 weeks, the reduced dose of HFA-BDP maintained equivalent lung function compared with all previous medications. Furthermore, asthma symptom scores revealed improvements across all groups and the proportion of symptom free days and nights and beta agonist free days increased significantly (p < 0.05) in all but one group. Quality of life improved with 80% of patients reporting goo/very good overall asthma control compared with 70% previously. Treatment-associated adverse events were generally infrequent, mild and transient. CONCLUSIONS: Patients on conventional inhaled corticosteroids may reduce their daily steroid dose to 800 microg or less whilst maintaining lung function and improving asthma symptom control by using the extrafine aerosol of HFA-BDP. PMID- 12911120 TI - Cefixime for the prophylaxis of urinary tract infections in children with malformative uropathies: an open study. AB - Urinary tract infections are often associated with urinary anomalies. An appropriate pharmacologic treatment may prevent, or may at least limit, any kidney damage due to pyelonephritis. The antibiotic prophylaxis plays a role as significant as early surgical therapy, taking into consideration also the present limitative trend for a softer therapeutic regimen. In the past few years a greater bacterial resistance has emerged against some commonly administered antibiotics. Cefixime (3rd generation cephalosporin) has been used on a wide series of patients suffering from urinary infections associated with urinary tract anomalies. A few significative results emerge from the present study. In conclusion, cefixime's effectiveness long-term prophylaxis of urinary infections associated with anomalies. PMID- 12911121 TI - Stem cell therapies in reparative medicine. AB - The future implementation of stem cell therapies to treat conditions thus far considered incurable has been envisioned as logical consequence of the fast-paced progress in stem cell research over the last few years. Still, many practical obstacles stand in the way to the routine application of these novel technologies in medicine. The conference "Stem Cell Therapies in Reparative Medicine," held aboard the cruise vessel Majesty of the Seas (Miami, USA-Nassau, Bahamas, April 19-22, 2002), focused on the analysis of these problems from different perspectives, including developmental biology (cell proliferation, fate determination, and enrichment), immunology (allorejection and prevention of autoimmunity recurrence), and clinical therapy, emphasizing the impact of stem cell technologies on the emerging field of tissue engineering and the treatment of alpha-1 antitrypsin deficiency. PMID- 12911122 TI - The testicular-derived Sertoli cell: cellular immunoscience to enable transplantation. AB - There is a renewed enthusiasm for the potential of cellular transplantation as a therapy for numerous clinical disorders. The revived interest is largely due to the unprecedented success of the "Edmonton protocol," which produced a 100% cure rate for type I diabetics following the transplantation of human islet allografts together with a modified immunosuppressive regimen. While these data provide a clear and unequivocal demonstration that transplantation is a viable treatment strategy, the shortage of suitable donor tissue together with the debilitating consequences of lifelong immunosuppression necessitate a concerted effort to develop novel means to enable transplantation on a widespread basis. This review outlines the use of Sertoli cells to provide local immunoprotection to cografted discordant cells, including those from xenogeneic sources. Sertoli cells are normally found in the testes where one of their functions is to provide local immunologic protection to developing germ cells. Isolated Sertoli cells 1) engraft and self-protect when transplanted into allogeneic and xenogeneic environments, 2) protect cografted allogeneic and xenogeneic cells from immune destruction, 3) protect islet grafts to reverse diabetes in animal models, 4) enable survival and function of cografted foreign dopaminergic neurons in rodent models of Parkinson's disease (PD), and 5) promote regeneration of damaged striatal dopaminergic circuitry in those same PD models. These benefits are discussed in the context of several potential underlying biological mechanisms. While the majority of work to date has focused on Sertoli cells to facilitate transplantation for diabetes and PD, the generalized ability of these unique cells to potently suppress the local immune environment opens additional clinical possibilities. PMID- 12911123 TI - Evaluation of different types of alginate microcapsules as bioreactors for producing endostatin. AB - The use of nonautologous cell lines producing a therapeutic substance encapsulated within alginate microcapsules could be an alternative way of treating different diseases in a cost-effective way. Malignant brain tumors have been proposed to be treated locally using engineered cells secreting proteins with therapeutic potential encapsulated within alginate microcapsules. Optimization of the alginate capsule bioreactors is needed before this treatment can be a reality. Recently, we have demonstrated that alginate-poly-L-lysine microcapsules made with high-G alginate and a gelled core disintegrated as cells proliferated. In this study we examined the growth and endostatin secretion of 293-EBNA (293 endo) cells encapsulated in six different alginate microcapsules made with native high-G alginate or enzymatically tailored alginate. Stability studies using an osmotic pressure test showed that alginate-poly-L-lysine alginate microcapsules made with enzymatically tailored alginate was mechanically stronger than alginate capsules made with native high-G alginate. Growth studies showed that the proliferation of 293 endo cells was diminished in microcapsules made with enzymatically tailored alginate and gelled in a barium solution. Secretion of endostatin was detected in lower amounts from the enzymatically tailored alginate microcapsules compared with the native alginate microcapsules. The stability of the alginate microcapsules diminished as the 293 endo cells grew inside the capsules, while empty alginate microcapsules remained stable. By using microcapsules made of fluorescenamine-labeled alginate it was clearly visualized that cells perforated the alginate microcapsules as they grew, destroying the alginate network. Soluble fluorescence-labeled alginate was taken up by the 293 endo cells, while alginate was not detected in live spheroids within fluorescence labeled alginate microcapsules. Despite that increased stability was achieved by using enzymatically tailored alginate, the cell proliferation destroyed the alginate microcapsules with time. It is therefore necessary to use cell lines that have properties more suited for alginate encapsulation before this technology can be used for therapy. PMID- 12911124 TI - Overexpression of telomerase confers a survival advantage through suppression of TRF1 gene expression while maintaining differentiation characteristics in K562 cells. AB - Leukemic stem cells that expressed endogenous telomerase activity were induced to show overexpression of exogenous hTERT and were analyzed for biological changes in order to assess the possible influence of telomerase gene therapy on the transplantation of normal hematopoietic stem cells. Introduction of hTERT into K562, a telomerase-positive immortal cell line, resulted in a 2.5-fold elevation of telomerase activity and the lengthening of telomeres by 6 kb to 23 kb. Real time fluorescent PCR, which could perform quantitative analysis of transcripts, revealed a 175-fold increase in hTERT expression, suggesting the posttranscriptional regulation of telomerase. Ectopic expression of hTERT in K562 cells showed a survival advantage during culture in the absence of serum. Expression of mRNA for the telomeric-repeat binding factor 1 (TRF1) and caspase-3 activity were both decreased in hTERT-transfected K562 cells. Transduced cells retained their usual phenotypic characteristics, differentiation ability, and signal transduction response to TPA. These data suggest that ectopic expression of hTERT by normal hematopoietic stem cells may confer a survival advantage without changing their innate biological characteristics. PMID- 12911125 TI - Suppression of allogeneic response by viral IL-10 gene transfer. AB - Th1 cell activation and cytokine production shift the balance between Th1 and Th2, favoring the upregulation of proinflammatory activity that leads to destruction of allogeneic hepatocytes following transplantation. Th2-type cytokines. such as IL-10, have immune regulatory function. The aim of this study was to determine the antirejection efficacy of allogeneic hepatocytes with spheroidal shape (spheroids) genetically modified with viral IL-10 (vIL-10). Allogeneic hepatocyte spheroids, transferred vIL-10 gene by using adenovirus as the vector, were transplanted into the spleen of Nagase's analbuminemic rats (NAR). NAR transplanted with vIL-10-transfected hepatocytes showed an abrupt rise in serum albumin levels that peaked on day 7 and remained at high levels up to day 21 after transplantation. The peak level of albumin on day 7 in vIL-10 transfected NAR was eminently higher than that in nontransfected NAR. Histopathological analysis revealed that in nontransfected NAR hepatocyte spheroids were more or less rejected on day 4, and, in contrast, vIL-10 transfected spheroids were still not rejected on day 14. This protective effect correlated with sustained high vIL-10 level in the splenic vein in NAR transplanted with vIL-10-transfected hepatocyte spheroids, suggesting that vIL-10 secreted from the transplanted hepatocytes induced an active suppression of allogeneic response. This study provides evidence to support the possibility of using vIL-10 gene therapy to prevent allogeneic response in hepatocyte transplantation. PMID- 12911126 TI - Transplants of rat chondrocytes evoke strong humoral response against chondrocyte associated antigen in rabbits. AB - Rat chondrocytes transplanted intramuscularly in rabbits produced cartilage. In 1 day-old transplants chondrocytes remained viable. After 1 week peripheral chondrocytes of the transplant were dead and the cartilage was surrounded and resorbed by macrophages. In 2-week-old transplants cartilage deteriorated and was invaded by fibroblast-like cells and macrophages. Sera of rabbits that received two or three consecutive transplants of rat chondrocytes with 2-week intervals contained high titer of antichondrocyte cytotoxic antibodies. A part of the cytotoxic activity could be removed by absorption with rat splenocytes. Western blot analysis of lysates from fresh or 24-h cultured chondrocytes with absorbed sera detected antigen with M(r), of approximately 74 and approximately 23 kDa. Only the latter remained after reduction in 2-mercaptoethanol. In lysates of fibroblasts and endotheliocytes the 23-kDa antigen was not found but the serum reacted with M(r) 39-kDa antigen. In lysates of thymocytes a weak band corresponding to M(r) of 35 kDa was present. Serum from rabbits receiving transplants of living chondrocytes followed by chondrocytes suspended in complete Freund's adjuvant contained antibodies directed against components of crude collagenase used for cell isolation. Such antibodies could not be detected in sera of rabbits receiving transplants of living chondrocytes only. Molecular weight of detected antigen differs from that of collagen type II, core of aggrecan, link proteins, and several other macromolecules of cartilage matrix. It could represent either a component of chondrocyte membrane or a membrane-bound substance resistant to enzymes used for isolation. Availability of antibodies against presumably chondrocyte-specific antigen produced during transplant rejection may help to characterize it more precisely and to ascertain whether its presence may influence results of autogenous chondrocyte transplants in humans. PMID- 12911127 TI - Fast hematopoietic recovery after bone marrow engraftment needs physiological proximity of stromal and stem cells. AB - Relatively slow hematopoietic recovery after isolated bone marrow (I-BM) engraftment is probably caused by a disrupted microenvironment of stromal and stem cells. Thus, we compared the kinetics of hematopoietic recovery of lethally irradiated rats that received I-BM versus vascularized BM (V-BM). Total body irradiated (TBI; 8 Gy) Lewis (LEW; RT1(1)) rats were either injected IV with syngeneic sex-mismatched 80 x 10(6) I-BM or transplanted with 80 x 10(6) V-BM in orthotopic hind limb grafts. Ten days later, peripheral blood (PB) and mesenteric lymph nodes (MLN) of these recipients were examined for the presence of donor derived hematopoietic cells with a panel of monoclonal antibodies by FACS. To detect male cells in sex-mismatched female recipients, PCR was performed using male Y chromosome primers. When examined in PB and MLN, recipients transplanted with V-BM displayed significantly faster recovery of leukocytes (CD43+), monocytes (CD14+), and T cells (CD5+) in comparison with I-BM recipients. In addition, only V-BM (but not I-BM) groups contained stroma-like male-positive cells in PB and MLN. Our results suggest that V-BM transplants provided superior hematopoietic recovery in comparison to I-BM transplants. We postulated that close proximity between stromal and stem cells in V-BM is essential for efficient repopulation with progenitors of different lines of leukocytes. PMID- 12911128 TI - Improved survival of macroencapsulated islets of Langerhans by preimplantation of the immunoisolating device: a morphometric study. AB - Encapsulation of cells in a semipermeable membrane may in the future provide an opportunity to treat a variety of endocrine and neurological disorders, without the need for lifelong immunosuppression. The physiological conditions in the device are crucial factors for graft survival. Previously, we have shown that the exchange across the immunoisolating membrane and the microcirculation around the TheraCyte device increase around 3 months after implantation. The aim of this study was to determine whether preimplantation of the TheraCyte device would improve the survival of a later transplanted islet graft. A TheraCyte device was implanted SC on one side of the back of a nondiabetic SD rat. After 3 months, 1500 islets isolated from SD rats were transplanted via the device port. At the same time, another device, loaded with the same number of islets, was implanted on the other side of the back. Both devices were explanted 2 weeks after islet transplantation (i.e., 3.5 months and 0.5 month after device implantation, respectively). Six pairs of devices were evaluated by morphometery. The volume densities of viable islets were 0.22 +/- 0.04 in the preimplanted device vs. 0.06 +/- 0.03 in the nonpreimplanted one (p < 0.05). The corresponding volume densities of fibrosis and necrosis were 0.64 +/- 0.13 vs. 0.85 +/- 0.08 (p < 0.05) and 0.11 +/- 0.14 vs. 0.09 +/- 0.07 (ns), respectively. When the absolute volumes (mm3) were calculated, preimplanted devices contained 1.1 +/- 0.7 endocrine cells while nonpreimplanted ones contained 0.4 +/- 0.2 (p < 0.05). The percentages of insulin- positive beta-cells in the preimplanted versus nonpreimplanted device were 80 +/- 5% and 67 +/- 6%, respectively (p < 0.01). The corresponding volumes of fibrotic tissue were 3.0 +/- 1.8 vs. 5.2 +/- 1.2 (p < 0.05), while the amount of necrotic tissue did not differ significantly (0.42 +/- 0.5 vs. 0.50 +/- 0.3). Preimplantation of the TheraCyte device seems to improve the survival of an encapsulated islet graft and reduce fibroblast outgrowth in the device. PMID- 12911129 TI - De novo liver tissue formation in rats using a novel collagen-polypropylene scaffold. AB - In experimental and clinical settings hepatocyte transplantation has provided limited benefit to patients with chronic liver disease because the transplanted hepatocytes were short-lived and were merely maintained for a brief period within the body. Except for whole-liver transplantation, creation of de novo liver tissue is necessary to treat this condition on a long-term basis. The aim of this study was to facilitate the formation of new tissue by actual self-regeneration, rather than by compensatory hypertrophy, or scar formation, with our collagen polypropylene composite scaffold. Collagen-polypropylene composite scaffolds, not containing hepatocytes, were implanted into the median liver lobe and the dynamics of new liver tissue formation was analyzed immunohistochemically over a 6-month period. Control scaffolds consisted of polypropylene scaffolds without collagen matrix. The control scaffold implants remained hollow throughout the study period and became encapsulated with a hard connective tissue capsule 1 week after implantation. In contrast, the collagen-polypropylene composite scaffold was filled with regenerating tissue structures 3 weeks after implantation. At this time, the predominant cell type within the scaffold was sesmin-positive stellate cells. A week earlier, oval cells were identified using monoclonal antibody staining (OV-6). Subsequently, these cells differentiated into alpha fetoprotein-positive immature hepatocytes. After 6 months, mature liver tissue, juxtaposed with bile ducts and blood vessels, was seen within the polypropylene scaffolds. We report the first evidence of de novo formation of liver tissue within a polypropylene scaffold, following implantation in the liver. This scaffold may play a role in treating chronic liver diseases requiring organ replacement therapy. PMID- 12911130 TI - Evaluation of 2-year-old intrasplenic fetal liver tissue transplants in rats. AB - Liver cell transplantation into host organs like the spleen may possibly provide a temporary relief after extensive liver resection or severe liver disease or may enable treatment of an enzyme deficiency. With time, however, dedifferentiation or malignant transformation of the ectopically transplanted cells may be possible. Thus, in the present study syngenic fetal liver tissue suspensions were transplanted into the spleen of adult male rats and evaluated 2 years thereafter in comparison to orthotopic livers for histopathological changes and (as markers for preneoplastic transformation) for cytochrome P450 (P450) and glutathione S transferase (GST) isoform expression. Because inducibility of P450 and GST isoforms may be changed in preneoplastic foci, prior to sacrifice animals were additionally treated either with beta-naphthoflavone, phenobarbital, dexamethasone, or the respective solvent. In the 2-year-old grafts more than 70% of the spleen mass was occupied by the transplant. The transplanted hepatocytes were arranged in cord-like structures. Also few bile ducts were present. Morphologically, no signs of malignancy were visible. With all rats, transplant recipients as well as controls, however, discrete nodular structures were seen in the livers. Due to age, both livers and transplants displayed only a low P450 2B1 and 3A2 and GST class alpha and mu isoform expression. No immunostaining for P450 1A1 was visible. At both sites, beta-naphthoflavone, phenobarbital, or dexamethasone treatment enhanced P450 1A1, P450 2B1 and 3A2, or P450 3A2 expression, respectively. No immunostaining for GST class pi isoforms was seen in the transplants. The livers of both transplant recipients and control rats, however, displayed GST pi-positive foci, corresponding to the nodular structures seen histomorphologically. Compared to the surrounding tissue, these foci also exhibited a more pronounced staining for GST class alpha and mu isoforms and a stronger inducibility of the P450 1A1 expression due to beta-naphthoflavone. In conclusion, in contrast to the livers, no preneoplastic foci seem to appear in the intrasplenic transplants even 2 years after transplantation. This may be due either to the protection of these transplants by the orthotopic livers or to the different humoral and nerval influences at the ectopic site. PMID- 12911131 TI - Effect of adhesion or collagen molecules on cell attachment, insulin secretion, and glucose responsiveness in the cultured adult porcine endocrine pancreas: a preliminary study. AB - The effect of either adhesion or collagen molecules on cell attachment, insulin secretion, and glucose responsiveness was investigated in adult porcine pancreatic endocrine (PE) cells that were cultured for a longer term in vitro. Six different types of molecules--laminin, fibronectin, poly-L-lysine (PLL), type I collagen, gelatin, and Matrigel--were used. Approximately 2.0 x 10(5) cells per dish of each molecule type were cultured for 4 weeks. In the laminin group, the insulin accumulation was maintained at a significantly higher level than in the control group at 4 weeks of culture, and glucose-stimulated insulin secretion and the insulin-positive rate were also higher than in the control group. In the Matrigel group, islet-like cell clusters were formed, but insulin accumulation rapidly decreased at 3-4 weeks of culture. A large number of PE cells attached tightly and spread in the fibronectin group until the fourth week of culture, but their function was not better than those in the control group. In the PLL and gelatin groups, the PE cell function was not significantly different from that of the control group. In the type I collagen group, insulin secretion was inferior to that of the other groups. The results of this study suggest that laminin is the most suitable extracellular matrix for the long-term culture preservation of PE cells. PMID- 12911132 TI - G-CSF-Mobilized human peripheral blood for transplantation therapy in stroke. PMID- 12911133 TI - Mobilized peripheral blood cells administered intravenously produce functional recovery in stroke. AB - Filgratism (granulocyte colony stimulating factor, G-CSF)-mobilized peripheral blood progenitor cells (PBPCs) have replaced bone marrow (BM) as a preferred source of autologous stem cells, in light of the faster hematologic recovery and lesser supportive care requirement exhibited by PBPC transplants. Other hematopoietic stem cells, like the human umbilical cord blood-derived stem cells (hUCBs), and nonhematopoietic stem cells have been shown to improve motor function in rodent models of injury and degenerative disease. In the present study we transplanted either G-CSF-mobilized PBPCs or hUCBs in rats 24 h after permanent middle cerebral artery occlusion (MCAO), and assessed their behavioral abnormalities in spontaneous activity and spontaneous motor asymmetry. In both transplanted groups of rats we observed a significant reduction of the stroke induced hyperactivity compared with nontransplanted, stroked animals. In addition, transplantation of G-CSF PBPC and hUCB cells prevented the development of extensive motor asymmetry. Our findings raise the possibility that PBPCs could provide a novel transplantation therapy to treat stroke. PMID- 12911134 TI - From James Parkinson to Friederich Lewy: leaving landmarks for further research journeys. PMID- 12911135 TI - IX Antonio Arrigo Award. Clinical neurophysiology in neuromuscular disorders: usefulness and limitations. AB - This lecture focuses on the impact of classic and modern electrophysiological techniques on the diagnostic approach to patients with neuromuscular disorders, placing emphasis on the limitations of currently used techniques and on the future prospects of clinical neurophysiology. The refinement of old techniques together with the development of new ones will hopefully improve the diagnostic capability of neurophysiology, improve understanding of the complex pathogenic mechanisms underlying neuromuscular disorders and help further the research of new therapeutic strategies. PMID- 12911136 TI - Aicardi-Goutieres syndrome: differential diagnosis and aetiopathogenesis. AB - Aicardi-Goutieres syndrome (AGS) is a progressive encephalopathy with onset in the first year of life and a recessive autosomal pattern of inheritance. The syndrome is characterised by acquired microcephaly, basal ganglia calcifications, white matter abnormalities, chronic cerebrospinal fluid (CSF) lymphocytosis and raised interferon-alpha (INF-alpha) in the CSF. AGS is diagnosed on the basis of a clinical picture characterised by microcephaly and by the onset of encephalopathy associated with severe psychomotor delay, spasticity and extrapyramidal signs. CT is very important in the diagnosis of AGS, demonstrating clearly the presence of calcifications at basal ganglia level: these are often bilateral and symmetrical. CT scan and MRI reveal leukodystrophy and progressive cerebral atrophy. A raised level of INF-alpha in the CSF constitutes a marker of the syndrome: this level, which falls with age, is higher in the CSF than in the serum, suggesting intrathecal synthesis. Differential diagnosis in AGS is carried out to exclude the presence of other neurological and endocrinological pathologies characterised by the presence of intracranial calcification; considering the white matter abnormalities, it is necessary to exclude forms of leukodystrophy associated with metabolic defects, known or otherwise. One fundamental aspect that remains to be clarified is the aetiopathogenetic mechanism underlying AGS: the most well-founded hypotheses are reported. There does not exist, to date, any causal therapy for AGS, although genetic studies, particularly those focusing on interferon-regulating genes, may well provide some therapeutic indications. PMID- 12911137 TI - Circadian organization of serum electrolytes in physiological aging. AB - Age-related structural and neurochemical changes occurring in the central nervous system have been related to changes in some rhythmometric parameters. In spite of their clinical importance, only a few studies have investigated the modifications over time of serum electrolytes in senescence. The aim of our study was to evaluate the circadian pattern of serum potassium, chloride, sodium, calcium and phosphorus in 30 clinically healthy elderly subjects, with no cognitive impairment, and to compare the findings with those given by 24 healthy young controls. The subjects were synchronized as regards their daily activities, sleeping/waking habits, time/quality of meals and dietary electrolyte intake. After an overnight fast, samples were taken beginning at 08.00 and every 4 h thereafter until 20.00, and every 2 h from 20.00 to 04.00. Both the young and the elderly subjects exhibited statistically significant circadian rhythms for all serum electrolytes considered. Our findings suggest that circadian organization of serum electrolytes is maintained in physiological aging, even though it should be noted that sodium and phosphorus acrophases differed significantly in the two experimental groups. PMID- 12911138 TI - The relationship between anaerobic lactate threshold and plasma catecholamines during incremental exercise in hereditary spastic paraplegia. AB - Lower limb muscle chronic hyperactivity in hereditary spastic paraplegia (HSP) is the consequence of motor corticospinal tract involvement, which in turn has been hypothesized to be of mitochondrial origin. In order to assess skeletal muscle aerobic metabolism and sympathetic response during exercise in 10 HSP patients, we evaluated their blood lactate and catecholamine levels during an incremental workload bicycle exercise. Lactate, but not epinephrine or norepinephrine, levels were significantly higher in the HSP patients than in control subjects, in both resting conditions and during exercise. In the patients, the anaerobic lactate threshold was reached prematurely (at 50% of the predicted normal maximal power output) when compared to normal controls. This finding was not related to any specific muscle morphology or histochemical activity. Although other factors, including chronic spasticity and muscle deconditioning, have to be considered in the interpretation of our data, our results suggest the possible involvement of a mitochondrial mechanism, independently of sympathetic system overactivation, in exercising skeletal muscle of HSP patients. PMID- 12911139 TI - Stereotyped movements in a group of autistic children. AB - The authors studied the stereotyped movements presented by a group of 20 autistic children, evaluating the patient observation protocols according to a psychodynamic model of autism. The stereotyped movements were analysed considering ten different parameters: type, site, morphology, frequency, association, complexity, active sensory channel, trigger event, affect during stereotyped movement, and function. The results indicate that each autistic child possesses highly idiosyncratic "stereotypical behavioural equipment", which includes behaviours ranging from simple, reflex-like actions, to much more complex movement patterns. Stereotyped movements were found to appear, in response to different stimuli, at moments when the subject's psychophysical state needed "resetting". Eight possible functions of stereotyped movements were identified. These correspond to two main purposes: reinforcement or weakening of the autistic barrier. The need for rehabilitation is questioned, since not all stereotyped behaviours interfere with explorative and cognitive activities. PMID- 12911140 TI - Disappearance of haemorrhagic stroke-induced thalamic (central) pain following a further (contralateral ischaemic) stroke. AB - We report the case of a patient who, following a right thalamic haemorrhage, developed thalamic syndrome characterised by burning pain and hyperalgesia in the left side of the body. Three years later, following a further (contralateral ischaemic) stroke, she reported the complete disappearance of the pain and hyperalgesia. To our knowledge, this is the first described case of disappearance of thalamic syndrome following a second stroke, different in nature from and contralateral to the first. Various hypotheses, based on the nervous tracts and nuclei involved in pain processing, may be advanced to explain this occurrence. PMID- 12911141 TI - Dizziness and migraine: a causal relationship? AB - Both migraine and dizziness are very frequent complaints, but the comorbidity of the two disorders is higher than it might be expected to be on the basis of chance alone. This implies a possible causal relationship, but definite diagnostic criteria for migraine-related vertigo are still lacking. Very recent attempts in this direction have shown that migraine may be the third leading cause of vertigo and that migraine-related vertigo may be effectively treated. A review of the literature on this topic, which includes some preliminary data of our own, demonstrates the difficulty in pinpointing migraine-associated vertigo as a clearly-defined entity. However, there is a measure of agreement on a few points: the spells of vertigo occur in patients who habitually suffer from motion sickness, and who have a history of migraine, either without or with aura; the delay between migraine and vertigo onset may be several years; migraine-related vertigo may be described as rotatory and/or as a feeling of unsteadiness, and single spells can occur without any other accompanying symptoms, however, when spells do occur in association with headache, they usually precede it. The vertigo duration may be shorter or longer than that of the migraine aura since it ranges from a few seconds to a continuous condition of unsteadiness. PMID- 12911142 TI - Cluster analysis: significance, empty space, clustering tendency, non-uniformity. II--Empty Space index. AB - The here presented Empty Space index (ES) evaluates the fraction of the information space without experimental points, i.e. the space where the distance from an experimental point is significantly larger than the mean distance between the experimental points themselves. ES can be used to eliminate the ambiguity of the some clustering indexes, that aim to evaluate the separation of the data set in clusters, but these clustering indexes are really a mixed measure of clustering, of empty space (the empty space does not necessarily correspond to the break between clusters) and of the degree of uniformity of the objects. The ES index can be used also to correct the MST index, the clustering index based on the distribution of edge lengths in the minimum spanning tree connecting the objects. The corrected MST index seems to be a reliable measure of the clustering degree. PMID- 12911143 TI - Binding properties of a polyclonal antibody directed towards lead complexes. AB - A previously described conjugate of 8-hydroxyquinoline and bovine serum albumin was complexed with lead(II), (8-hydroxyquinoline to metal ion ratio 2:1) and used as an immunogen to produce polyclonal antibodies against lead in chickens. Antibodies obtained from a first blood sample during a standard immunisation procedure showed very promising features (dynamic range of the assay was 1 to 1000 ng l(-1)). Nevertheless, proceeding with the immunisation caused a complete loss of the recognition of the complex. A modified brief immunisation procedure was carried out and, in this case, the immunogen proved to be sufficiently stable in vivo to produce antibodies that selectively bound to the lead(II) complex (in the same 2:1 ratio used as an immunogen). Since the antiserum obtained cannot reach the same performance levels as the first one, standard curves were obtained by adding the free 8-hydroxyquinoline to the solution, which enables 2:1 complexes to be more easily formed. Cross-reactivity and dependence from buffer were investigated, showing at least 10-fold lower binding to non-target divalent metal ions compared to lead(II). MES buffer (pH = 6.0) gave more sensitive but very imprecise curves, whereas Tris (pH = 8.5) allows higher precision but lower sensitivity to be observed. PMID- 12911144 TI - Lead (II) cholate solubility. AB - In the framework of the research carried out on the behaviour of the salts of bile acids in aqueous solutions, the lead (II) cholate solubility was determined at 25 degrees C and in 0.100, 0.500 and 0.800 mol dm(-3) N(CH3)4Cl, as ionic medium. The change of its solubility was studied as a function of the cholate and hydrogen ion concentration. Solubility and electromotive force measurements of suitable galvanic cells were carried out and from the results lead (II) cholate solubility product could be calculated and the presence of associated species in solution in the ratio 1:3 and 1:4 between lead (II) and cholate was assumed. The relative constants were determined, as well. The results of this work allow us to obtain the free cholate concentration in equilibrium with solid lead (II) cholate. PMID- 12911145 TI - Monitoring of an industrial process by multivariate control charts based on principal component analysis. AB - The control and monitoring of an industrial process is performed in this paper by the multivariate control charts. The process analysed consists of the bottling of the entire production of 1999 of the sparkling wine "Asti Spumante". This process is characterised by a great number of variables that can be treated with multivariate techniques. The monitoring of the process performed with classical Shewhart charts is very dangerous because they do not take into account the presence of functional relationships between the variables. The industrial process was firstly analysed by multivariate control charts based on Principal Component Analysis. This approach allowed the identification of problems in the process and of their causes. Successively, the SMART Charts (Simultaneous Scores Monitoring And Residual Tracking) were built in order to study the process in its whole. In spite of the successful identification of the presence of problems in the monitored process, the Smart chart did not allow an easy identification of the special causes of variation which casued the problems themselves. PMID- 12911146 TI - Optimization of an urban particulate matter multi-element analysis method by inductively coupled plasma--atomic emission spectrometry (ICP-AES). AB - In the present work a method for simultaneous metals determination, in urban air particulate matter by ICP-AES has been set up. A large number of elements (18) has been analyzed, including major (Al, Fe, K, and Mg), minor (Na, Pb and Zn) and trace (As, Cd, Co, Cr, Cu, Hg, Mn, Ni, Sb, Sr and V) elements. The procedure consists of microwave sample acidic total digestion by HNO3/HF mixture and subsequent analysis by ICP-AES, using different assemblies depending on sample treatment procedure: a quartz Meinhard nebulizer/cyclonic chamber, if HF excess was eliminated, or a cross-flow nebulizer/plastic Scott chamber, suitable for application with HF. A cyclonic chamber for hydride generation was used for As, Sb and Hg determination. The procedure was tested with Standard Reference Materials 1648 NIST Urban Particulate Matter and Certified Reference Material No8 NIES "Vehicle Exhaust Particulates". Two sampling supports, quartz fibre and polycarbonate filters, have been examined in order to find the most suitable i.e. the one characterized by less interference. Some real samples of urban air particulate matter, TSP, PM10 and PM2.5 fractions, collected during an intercomparison campaign promoted by Regione Lombardia, have been analyzed with the procedure developed. PMID- 12911147 TI - Neural network modelling of an electrochemical process. AB - Artificial Neural Network (ANN) methodology has gained popularity in chemistry in recent years as a result of its ability to solve problems for various purposes. In particular, ANN shows a very high performance in the modelling of the experimental measurements. To this aim, the trained ANN is used to predict unknown values of measurement system. For a given trial set of parameters, the experimental response may be predicted by the model. In recent decades, several investigations were based on the electrooxidation of D-glucose owing to its many applications such as detection systems (glucose sensors), fuel cells and synthesis of economically interesting products. In the present work based on the electrochemical process modelling, the estimation of peak current densities of the D-glucose electrooxidation on palladium electrode in alkaline medium was investigated as a function of potential sweep rate and D-glucose concentration by using a three layer feed-forward ANN with error propagation learning algorithm. PMID- 12911149 TI - Environmental monitoring for microbial contamination by batch and flow bioluminescent systems. AB - A study was performed to optimise a rapid method to determine the microbial content on surfaces by means of flow and batch microplate ATP bioluminescent assay. Sampling, ATP extraction and testing of these specimens were considered. The data obtained gave a general picture of the state of hygiene and cleanliness of the surfaces examined, mainly classrooms. PMID- 12911148 TI - Selective determination of trace copper(II) by cathodic adsorptive stripping voltammetry with a naphthol-derivative Schiff's base. AB - A selective and sensitive stripping voltammetric method for the determination of trace amounts of copper(II) with a recently synthesized naphthol-derivative Schiff's base (2,2'-[1,2-ethanediylbis(nitriloethylidyne)]bis(1-naphthalene)) is presented. The method is based on adsorptive accumulation of the resulting copper Schiff's base complex on a hanging mercury drop electrode, followed by the stripping voltammetric measurement at the reduction current of adsorbed complex at -0.15 V (vs. Ag/AgCl). The optimal conditions for the stripping analysis of copper include pH 5.5 to 6.5, 8 microM Schiff's base and an accumulation potential of -0.05 V (vs. Ag/AgCI). The peak current is linearly proportional to the copper concentration over a range 2.3-50.8 ng ml(-1) with a limit of detection of 1.9 ng ml(-1). The accumulation time and RSD are 90 s and (3.2 3.5)%, respectively. The method was applied to the determination of copper in some analytical grade salts, tap water, human serum and sheep's liver. PMID- 12911150 TI - Uncertainty estimation in chemical measurements: carbon dioxide determination at atmospheric concentration. AB - The present work deals with the evaluation of measurement uncertainty in the determination of carbon dioxide (CO2) concentration in atmosphere, given the high relevance of this greenhouse gas that influences earth climate. In order to carry out CO2 measurements, non dispersive infrared (NDIR) analysers are usually employed as they are stable and scarcely affected by interferences from other air components or pollutants. Typical uncertainty sources are the resolution of the analyser, its time drift and the contributions due to instrument calibration, which is required in order to produce traceable measurement results. The calibration uncertainty takes into account the uncertainty of the composition of the calibration gas mixtures, the instrument repeatability and the possible or residual lack of fit of the adopted mathematical model. PMID- 12911151 TI - Interpretation of atmospheric pollution phenomena in relationship with the vertical atmospheric remixing by means of natural radioactivity measurements (radon) of particulate matter. AB - In this paper the results of seasonal monitoring campaign for primary (benzene and carbon monoxide) and secondary (nitrogen dioxide and ozone) pollutants and for the natural radioactivity of the particulate matter in the urban area of Rome, are reported to investigate acute atmospheric pollution episodes. Comparing the daily concentration trends of primary and secondary pollutants with those of the natural radioactivity, considered as index of the vertical diffusion in the low boundary layer, it has been evidenced that the acute pollution episodes in Rome occur in the winter period for the high atmospheric stability (primary pollution) and in the summer period for the strong diurnal atmospheric remixing (secondary pollution). PMID- 12911152 TI - Experimental study of the pH influence on the transport mechanisms of phenols in soil. AB - The study of the transport mechanisms connected to solid-liquid interactions is fundamental in the determination of the extension of the pollution of a site and in the evaluation of the best remediation process to be applied. The sorption of hydrophobic ionizable organic contaminants from the groundwaters is supervised not only by the physico-chemical properties of soil and pollutants, but also by the groundwaters pH, which deeply influences their solubility in the aqueous media, and consequently their transport mechanisms in the aquifer. In this work an experimental study of the sorption of phenol and 2-nitrophenol on two soils, different in particle-size distribution, CEC and organic carbon content, was realized. The sorption potential of the soils was evaluated by means of a physical, mineralogical and chemical characterization. The experimental data coming from some batch tests, performed at pH values equal to 4, 7 and 10 were fitted by means of linear, Freundlich and Langmuir isotherms models. The soil contaminants interaction mechanisms that influence the isotherms shapes were then analyzed and discussed, and a comparison between the theoretical and experimental values of the partitioning coefficient KD was performed. PMID- 12911154 TI - The effects of interaction between environment and constituent materials of Palazzo San Michele in Fano (Pu). PMID- 12911153 TI - Elimination of the interferences of anions in the kinetic spectrophotometric determination of vanadium (V) solution. AB - Accurate determination of vanadium (V) in industrial waste water is of great importance in environmental, biological and toxicological studies. Most of kinetic spectrophotometric methods based on the catalytic effect of vanadium (V), when applied to real samples for determination of trace levels of vanadium (V) lack the satisfactory sensitivity and selectivity. This may be attributed to the serious interferences of various anions which are common pollutants in industrial waste water. The oxidation of gallic acid by ammonium persulphate, catalysed by vanadium (V) was chosen for our study. The effect of the serious interferences of various anions such as chloride, bromate, bromide, chromate, iodide, iodate, molybdate, carbonate and sulphate on the net absorbance given by 4 microg l(-1) of vanadium (V) solution were studied. The minimum concentrations of citric acid, EDTA, ascorbic acid and oxalic acid as leveling off agents required to level off interfering effects due to the aforementioned anions in the kinetic determination of vanadium (V) were 50, 70, 80 and 120 microg ml(-1), respectively. In the presence of optimum concentrations of effective leveling off agents, the dynamic range can be extended and sensitivity increased as compared with the proposed method without levelling off agents. The proposed method is a rapid, sensitive and selective method for the determination of ultra trace amounts of vanadium (V) in real samples with satisfactory results. PMID- 12911155 TI - Acute uremia produced in rats by nephrotoxic chemicals is alleviated by protein deficient diet. AB - Rats injected with mercuric chloride develop an acute renal tubular necrosis with uremia, which is frequently lethal. Pretreatment for 3 or 7 days with a protein free diet reduces the mortality, the clinical signs (tremor), and the severity of renal tubular necrosis, and ameliorates the uremic chemical findings in the serum. Similar results followed injection of a nephrotoxic amino acid, D-serine, after pretreatment with a protein-free diet. Indirect evidence suggests that induction of metallothionein may be involved, at least in the experiments with mercury. Acute uremia produced by nephrotoxic chemicals may be useful for further studies of the role of nutrition in uremia, while avoiding the surgical procedures and prolonged observations required for the "remnant kidney" models. PMID- 12911156 TI - Transdermal delivery of chloroquine by amidated pectin hydrogel matrix patch in the rat. AB - The aim of the present study was to investigate the suitability of amidated pectin matrix patch for transdermal chloroquine delivery in an effort to mask the bitter taste when orally administered. Chloroquine has easily measurable outputs that are linked to increased renal Na+ excretion. We thus monitored urinary Na+ output in separate groups intravenously administered chloroquine or topically applied pectin hydrogel chloroquine matrix patch. Male groups of anesthetized Sprague-Dawley rats were placed on a continuous jugular infusion of 0.077 M NaCl at 150 microL min(-1). After 3 h equilibration period, consecutive 20 min urine collections were made over the subsequent 4 h of 1 h control, 1 h 20 min treatment, and 1 h 40 min recovery periods for measurements of urine flow and Na+ and K+ excretion rates. The effects of intravenous chloroquine infusion or topical application of pectin hydrogel chloroquine matrix patch were examined in rats in which the drug was added to the infusate or patch applied onto the shaved area during the 1 h 20 min treatment period. The animals were switched back to the infusate alone for the final 1 h 40 min recovery period. Vehicle infused animals acted as controls. Trunk blood was collected after the treatment period from parallel groups for chloroquine measurements. The plasma chloroquine concentrations following iv chloroquine or application of pectin chloroquine hydrogel matrix patch were 9.3 +/- 0.8 mg L(-1) and 7.3 +/- 1.1 mg L(-1) respectively (n = 7 in both groups). Chloroquine infusion and pectin chloroquine patch significantly (p < 0.01) increased Na+ excretion to peak values of 14.1 +/- 0.9 micromol min(-1). and 20.35 +/- 1.0 micromol min(-1), respectively by comparison with controls (9.1 +/- 0.9 micromol min(-1)), at the corresponding period. The results suggest that the pectin chloroquine patch matrix preparation has potential applications for transdermal delivery of chloroquine and perhaps in the management of malaria. PMID- 12911157 TI - Evolution of total antioxidant status in a model of acute renal insufficiency in rats. AB - BACKGROUND: Accurate estimation of the Total Antioxidant Status (TAS) in the myoglobinuric acute renal failure (ARF) is necessary because its pathogenesis is believed to be mediated, at least in part, by the development of oxidative stress resulting from the generation of oxygen free radicals and reduced antioxidant defense system. The purpose of this study is to examine the TAS 24 and 72 h after glycerol injection in a model of myoglobinuric-ARF. EXPERIMENTAL DESIGN: The study was conduced in 28 Sprague-Dawley rats. In group 1 (n = 7) rats were placed into individual metabolic cages and deprived of water during 24 h. afterwards an intramuscular injection of glycerol was administrated (50% vol/vol in sterile saline) 10 mg/100 g of body weight and 24 h later blood samples were collected for biochemical measurements (urea, creatinine, creatine-kinase, and TAS levels). In group 2 (n = 7), rats followed the same conditions than group 1 ones but blood samples were collected 72 h after glycerol injection. In groups 3 (n = 7) and 4 (n = 7) rats didn't receive glycerol injection, and blood samples were collected within 24 and 72 h respectively after they were placed into metabolic cages. RESULTS: In groups 1 and 2 we observed a renal function decrease, with higher serum levels of urea and creatinine in group 2 (urea levels: 269 +/- 16 mg/dL vs. 586 +/- 147 mg/dL; p < 0.001. Creatinine levels: 2.8 +/- 0.2 mg/dL vs. 5.8 +/- 0.7 mg/dL; p < 0.001). TAS levels in groups 2, 3, and 4 were similar, but in group 1 was significantly lower (group 1: 0.81 +/- 0.2 mmol/L; group 2: 1.3 +/- 0.1 mmol/L; group 3: 1.2 +/- 0.3 mmol/L, and group 4: 1.2 +/- 0.2 mmol/L; p < 0.005). CONCLUSION: In the model of glycerol induced myoglobinuric-ARF we observed a decrease of serum TAS level within 24 h with spontaneous recuperation 72 h after. PMID- 12911158 TI - The effect of intense passive apnea in the acid-base balance and electrolytes. AB - The aim of this study was to investigate the disturbances of the acid-base balance and electrolyte disorders in healthy individuals in the greatest passive apnea (prior to apnea and prior to first inhalation). The changes of the acid base balance, the serum potassium and calcium levels as well as the disturbances of the cardiac rhythm were studied in a group of 15 healthy males aged 18 to 41 years old. Seven individuals part of the original group participated in a second trial. Following the initial apnea trial the individuals were allowed to 2-3 free inhalations and a second passive apnea effort was performed. The same parameters were measured in the second effort. In the primary effort the apnea duration was 170 +/- 53 s (mean +/- SD). The results received prior to the effort and immediately before the end of the apnea (paired t-test), showed no significant changes in the blood pH. The PaCO2 increased significantly, the PaO2 decreased significantly, blood bicarbonate increased significantly, the hemoglobin oxygen saturation decreased significantly. In addition the serum potassium and calcium levels decreased significantly. In the subgroup (7 individuals) the second effort of apnea lasted 50-70 s and showed no significant changes in the blood pH. The same changes concerning PaCO2 and blood bicarbonate were observed in the first and second effort of passive apnea. The serum potassium and calcium levels as well as the hemoglobin oxygen saturation showed no significant changes. During the initial effort of apnea all men presented bradycardia, which was followed by tachycardia immediately after the end of the effort. One individual, whose effort lasted 290s suffered intense tachycardia, cyanosis, convulsions, and loss of consciousness. It is concluded that during an intense effort of passive apnea: (a) the blood PaCO2 is significantly increased, as well as the blood bicarbonates, (b) the PaO2, the hemoglobin oxygen saturation and the serum potassium and calcium levels are significantly decreased, (c) the cardiac rate is significantly decreased, and (d) the second effort of apnea, following immediately the initial one, is sorter in duration and produces similar or milder changes of the above parameters. PMID- 12911159 TI - Risk factors for development of acute renal failure after liver transplantation. AB - BACKGROUND: Acute renal failure (ARF) is a common complication after liver transplantation (LTx). Identification of risk factors may prevent the development and attenuate the impact of ARF on patients outcome after LTX. METHODS: Retrospective analysis of variables in the pre, intra, and postoperative periods of 92 patients submitted to LTx was performed in order to identify risk factors for development of ARF after LTx. ARF was defined as serum creatinine > or = 2.0 mg/dL in the first 30 days after LTx. Univariate and multivariate analysis by logistic regression were performed. RESULTS: ARF group comprised 56 patients (61%). Preoperative serum creatinine was higher in ARF group. During the intraoperative period, ARF group required more blood transfusions, developed more episodes of hypotension and presented longer anesthesia time. In the postoperative period, ARF group presented higher serum bilirubin and more episodes of hypotension. Dialysis was required in 10 patients (11%). The identifled risk factors for development of ARF were: preoperative serum creatinine > 1.0 mg/dL. more than five blood transfusions in the intraoperative period, hypotension during intra and postoperative periods. The identified mortality risk factors were hypotension in the postoperative period and no recovery of renal function after 30 days. CONCLUSIONS: Several factors are involved in the pathogenesis of ARF after LTx and may influence patients outcome and mortality. Pretransplant renal function and hemodynamic conditions in the operative and postoperative periods were identified as risk factors for development of ARF after LTx. Nonrenal function recovery and postoperative hypotension were identified as mortality risk factors after LTx. PMID- 12911160 TI - Urinary transforming growth factor-beta1 excretion in renal allograft recipients during the early post-transplantation period. AB - BACKGROUND: Transforming growth factor-beta1 (TGF-beta1), the major fibrogenic growth factor, is implicated in the pathogenesis of renal scarring in experimental and clinical nephropathies as well as in chronic allograft nephropathy. In this study we examined the pattern of changes of TGF-beta1 excretion in the urine and the sites of TGF-beta1 expression in the kidney of transplanted patients during the early post-transplantation period. METHODS: Eighteen renal allograft recipients were included in the study. In all patients urinary TGF-beta1 levels were determined by ELISA in sequential measurements during the first two postoperative months and compared to that of 14 healthy subjects. The renal expression of TGF-beta1 protein was studied in 4 patients that underwent a biopsy of the transplanted kidney at the same period. All patients were treated with prednisolone, cyclosporin, and mycophenolate mofetil. RESULTS: Urinary TGF-beta1 levels were increased during the first postoperative days. Although they were gradually reduced during the first two post-operative months, they remained significantly higher compared to those of normal subjects (580 +/- 148 ng/24 h vs. 310 +/- 140 ng/ 24 h p < 0.01). The decline of urinary TGF-beta1 excretion followed that of serum creatinine. TGF-beta1 protein expression was identified within the cytoplasm of tubular epithelial cells of transplanted patients. CONCLUSIONS: Elevated urinary TGF-beta1 levels are observed during the early post-transplantation period in renal allograft recipients and are maintained high even after restoration of renal function to normal. PMID- 12911161 TI - Increased endothelin: nitric oxide ratio is associated with erythropoietin induced hypertension in hemodialysis patients. AB - Regular administration of recombinant human erythropoietin (rHuEPO) is frequently associated with a rise in arterial blood pressure in hemodialysis (HD) patients. The aim of this study was to examine the effects of rHuEPO on plasma endothelin (ET)-1 and nitric oxide products (NOx) concentration in HD patients. Fifteen patients on maintenance HD with hematocrit of less than 25% were included in the present study. All patients received 3,000 units of rHuEPO intravenously three times a week at the end of each HD session. Plasma levels of ET-1, NOx, thromboxane B2 (TXB2), prostacyclin (6-keto-PGF1alpha), and cyclic guanosine 3',5'-monophosphate (cGMP) were measured before, 2, and 4 weeks after rHuEPO treatment. Plasma concentrations of ET-1, TXB2, and 6-keto-PGF1alpha were measured by radioimmunoassay. Plasma NOx was measured by high-performance liquid chromatography. An rHuEPO-induced increase in mean arterial blood pressure of over 6 mmHg occurred in 7 patients (hypertensive group), whereas the elevation of mean arterial blood pressure was less than 5 mmHg in 8 patients (nonhypertensive group). Plasma ET-1 levels were elevated in all HD patients. Elevated plasma ET-1 levels remained unchanged after rHuEPO treatment in the hypertensive group, whereas the increase in plasma ET-1 levels was attenuated in the nonhypertensive group. Plasma NOx concentrations were also increased in all HD patients. This increase in plasma NOx levels was lessened in the hypertensive group after rHuEPO administration; however, plasma NOx levels remained increased in the nonhypertensive group. Changes in mean arterial blood pressure were significantly correlated with changes in plasma ET-1/NOx ratio. Plasma levels of TXB2, 6-keto PGF1alpha, and cGMP were unchanged after rHuEPO administration in the hypertensive and nonhypertensive groups. These results suggest that an increase in ET-1/NOx ratio in blood, probably occurring in vascular endothelial cells, may be associated with rHuEPO-induced hypertension in HD patients. PMID- 12911162 TI - Seven-year study of bacteremia in hospitalized patients on chronic hemodialysis in a single tertiary hospital. AB - Sixty-seven hospitalized bacteremic patients on chronic hemodialysis were matched with 134 bacteremic controls. Mortality (25% and 28% respectively) was unrelated to dialysis. Predictors of mortality were: advanced age, decreased functional capacity, and malignancies. Most common organisms recovered were Staphylococcus aureus (30%), coagulase negative Staphylococcus (15%), and polymicrobial agents (15%). PMID- 12911163 TI - Serum alpha1-antitrypsin but not complement C3 and C4 predicts chronic inflammation in hemodialysis patients. AB - BACKGROUND: We studied whether predialysis serum levels of positive acute phase markers such as alpha1-antitrypsin (AT), and complement components C3 and C4 could identify the presence of chronic inflammation in maintenance hemodialysis (HD) patients. METHODS/RESULTS: In 103 stable HD patients, AT directly correlated with C-reactive protein (CRP) (P < 0.005), alpha1 acid-glycoprotein (P < 0.005), fibrinogen ( P < 0.05), lipoprotein (a) (P < 0.01) and von Willebrand factor antigen (P < 0.05), while C3 and C4 were not related to any of these inflammatory markers. In the patients with elevated CRP and hypoalbuminemia, the mean AT value of 1.74 +/- 0.50 g/L was higher (P = 0.008) than that of 1.38 +/- 0.27g/L in the subjects with normal CRP and albumin. Using the above cut-off levels, the positive and negative predictive values of AT on the presence of severe inflammation were 0.86 and 0.62, respectively, and the sensitivity and specificity were 86% and 73%, respectively. CONCLUSION: Serum AT levels above 1.74 g/L and below 1.38 g/L may select the HD patients with severe inflammation from those without. Measurements of C3 and C4 are not helpful in this situation. PMID- 12911164 TI - Secondary hyperparathyroidism in chronic hemodialysis patients: prevalence and race. AB - Secondary hyperparathyroidism is a common complication of renal failure. The exact prevalence in chronic hemodialysis patients in not known. We evaluated 122 patients who were receiving maintenance hemodialysis for at least 12 months in 2 dialysis centers in mid Michigan. Seventy-eight percent of the patients had iPTH above 200 pg/mL (mean 481 pg/mL), 19% had iPTH within the accepted normal range (mean 155 pg/mL), while 3% had level below 100 (mean 53 pg/mL). Phosphate, calcium, calcium phosphate product, age and time on dialysis are the important factors correlating with elevated iPTH. There was no significant difference in iPTH between diabetic and nondiabetic patients with mean iPTH of 403 pg/mL and 407 pg/mL respectively. Black patients had a statistically significant elevated iPTH compared with white patients with a mean iPTH of 438 pg/mL and 283 pg/mL respectively (p < or = 0.004). Factors that predict the response to vitamin D therapy need to be evaluated to help reduce the high prevalence of secondary hyperparathyroidism. The patterns of bone disease in black patients need to be evaluated to further define the accepted normal iPTH range for this population. PMID- 12911165 TI - Progression of renal disease measured by the reciprocal of serum creatinine. AB - BACKGROUND: This study aims to quantify the decrease in the reciprocal of serum creatinine during the two years prior to the start of dialysis and to identify the factors associated with the progression of renal disease. METHODS: The study included 76 patients who were monitored for an average of 26.27 months. During the study 52 patients began dialysis treatments. The clinical and analytical variables were analyzed using the t-student test, ANOVA or the Kruskall-Wallis test. A multiple regression analysis was carried out with the decrease in the reciprocal of serum creatinine as the dependent variable. RESULTS: There was a decrease between the mean value during the two years prior to dialysis, the mean value during the previous year and the value at the start of dialysis treatments of 0.29, 0.21 and 0.17 respectively. The nephropathy having the sharpest drop in the reciprocal of serum creatinine, expressed as a percentage of the initial value, was found to be nephroangiosclerosis (46.29%). A greater decline in the reciprocal of serum creatinine was exhibited by low albumin values and adjusted by means of different variables with LDL levels (p = 0.141). For different levels of haemoglobin, proteinuria and cholesterol, the drop in the reciprocal of serum creatinine was greater in patients having a diastolic blood pressure reading of 90 or greater. CONCLUSIONS: The decrease in the reciprocal of serum creatinine was associated with different variables in patients who required dialysis. PMID- 12911166 TI - Results of the pregnancies with HELLP syndrome. AB - In this study, clinical features, developing complications, and results of thirty six patients, which were followed up in our Obstetrics and Gynecology and Nephrology departments between 1997 and 2001, with the diagnosis of HELLP syndrome were searched retrospectively. The mean age of the cases followed up with diagnosis of HELLP syndrome were 30.2 +/- 5.9 (17-46) years. HELLP syndrome was diagnosed on average in the 32.6 +/- 4.8th (23-41) week of gestations. Seventy percent of the cases were with severe preeclampsia and 30% of the cases were with mild preeclampsia. Eleven cases (30%) were nullipara and twenty-five cases (70%) multipara. The average of arterial systolic blood pressure of the cases were 161.6 +/- 26 mmHg, and that of diastolic blood pressure was 98.5 +/- 16.8 mmHg. In thirteen cases (36%) acute renal failure (ARF), six cases (17%) placenta detachment, two cases disseminate intravascular coagulation (DIC), one case Adult Respiratory Distress Syndrome (ARDS) were developed. In seven cases (19%) intrauterine dead fetuses were detected. In twenty-three cases by cesarian section (64%), in thirteen cases by induction (36%) the pregnancies were terminated in 72 h after diagnosing HELLP syndrome. Birth weights of eleven babies (30%) were below 1500 g. Five of the eleven babies were dead in the neonatal period. Six of the thirteen patients who had ARF were given hemodialysis. Two patients died because of the development of ARF + DIC and ARDS. No predicting factors for the development of HELLP syndrome could be detected, but severe preeclampsia. Therefore we think that preeclamptic pregnancies must be followed up very closely and if HELLP syndrome develops, termination of the pregnancy would be proper as soon as possible. PMID- 12911168 TI - Risk factors for hospital death of patients with end-stage renal disease without previous diagnosis of severe chronic renal failure arriving in an emergency situation at the hospital. AB - OBJECTIVES: Many end stage renal disease (ESRD) patients get their first nephrologic care under critical clinical conditions and without previous diagnosis of chronic renal failure (CRF), a situation even worse than the late referral of CRF patients for nephrologic treatment. Data on these "nonreferred" patients are scarce. The objectives of this study were to assess clinical and laboratory features, the reasons for coming to the hospital and the factors associated with death in nonreferred ESRD patients first seen by a nephrologist in an emergency situation. METHODS: Retrospective study (April 1996-March 2000) using the medical records of patients diagnosed with ESRD at the nephrologic emergency visit in a university tertiary hospital. Clinical and laboratory parameters were reviewed. Patients were divided into two groups according to hospital outcome: survivors or nonsurvivors. RESULTS: There were 414 patients (12% of all nephrologic emergency visits), aged 49 +/- 17 years, 266 males (64%) and 208 (55%) hypertensive. Mortality rate was 13.7% (54/393). When compared to nonsurvivors were older, used mechanical ventilation and vasoactive drugs more frequently, presented higher infection rate, and showed lower plasma creatinine. Multivariate logistic regression showed as factors independently associated with death: first nephrologic visit at intensive care unit, infection as cause for seeking medical care, and increasing age. Plasma creatinine above 10 mg/dL was a protective factor for death. CONCLUSIONS: ESRD patients reaching dialysis in a nephrologic emergency situation presented high hospital mortality, which was mostly associated with their poor clinical condition at admission. PMID- 12911167 TI - Glomerulopathy in Kuwait: the spectrum over the past 7 years. AB - There are few studies that examine, prospectively, the epidemiological profile of glomerulopathy (GP) and its clinicopathological correlation. All patients referred to Al-Amiri renal center in Kuwait from January 1st, 1995 to December 31st, 2001 were screened for GP. Detailed clinical data were collected and serological markers were done. Renal biopsy was performed whenever indicated. During those 7 years, a total of 584 patients were diagnosed, on histological basis, to have GP, 315 of whom were Kuwaiti nationals. During the same period of the study, 26 patients presented with bilateral small kidneys, history of proteinuria > 2 g/day and lacked systemic manifestations of autoimmune disease. Furthermore, 164 patients with clinical manifestations of diabetic glomerulosclerosis were not subjected to kidney biopsy. Hence, the calculated annual incidence rate of GP in Kuwaiti nationals was 34.5 per 100,000 population (PTP). The calculated rate of diabetic glomerulosclerosis was 13.4 PTP and that of nondiabetic 21.1 PTP. The calculated incidence rates of GP increased with age and were twice as high in males compared to females. Vasculitis was more common in elderly males while SLE nephritis was a disease of adults, 88.7% of whom were females. In the subgroup of primary GP, focal segmental glomerulosclerosis was the most common histological lesion accounting for 18.0% of the total biopsies in Kuwaiti patients, yet only 36.8% of those who fulfilled the criteria of primary type. Minimal change disease was the second primary GP (13.0%), followed by immunoglobulin A deposition disease (7.9%) and membranous glomerulonephritis (5%). Autoimmune diseases such as systemic lupus erythematosus (SLE) and vasculitis were common. Interestingly, only 44 of 72 (61.1%) of patients with SLE and 11 of the 62 (17.7%) of patients with vasculitis presented with rapidly progressive glomerulonephritis. On the other hand, 10 of 58 (17.2%) patients with nephroangiosclerosis presented with renal failure and protein excretion > 2 g/day simulating primary GP. Furthermore, only 21 of 40 (52.5%) patients with IgA nephropathy presented with "benign disease". Prospective studies are essential to ascertain the actual incidence and etiology of GP. The loose clinicopathological correlation in GP dictates an aggressive diagnostic approach in its study and management. PMID- 12911169 TI - Reversible thick ascending limb dysfunction and aseptic meningitis syndrome: early manifestation in two leptospirosis patients. AB - Leptospirosis is a widespread spirochetal zoonosis caused by the members of the genus Leptospira. The natural history of human leptospiral infection varies widely. The infection can cause a subclinical illness, or may be mistaken for influenza. In individuals who become ill, leptospirosis typically presents as one of two clinically recognizable syndromes. The first syndrome is the mild anicteric form, which rarely results in death, while the second syndrome fulminant icteric form, known as Weil's syndrome, has an associated 10% mortality. The anicteric form comprises two disease stages, namely the septicemic phase and the immune phase. In fever work up, leptospirosis is usually not the first considered pathogen of sepsis, unless jaundice and ARF are present. This study investigated two patients with leptospirosis presenting with conscious disturbance and oligoric acute renal failure individually. In the second patient, persistent hypokalemia and metabolic alkalosis developed during recovery from acute renal failure. Several tubular function tests were performed to define the renal tubular lesion in this patient, revealing a defect on the thick ascending limb. This study also reviews previous studies on leptospirosis including its epidemiology, pathogenesis, clinical presentation, diagnosis, treatment, and prognosis. PMID- 12911170 TI - Ciprofloxacin-induced acute interstitial nephritis and autoimmune hemolytic anemia. AB - Ciprofloxacin has been associated with several side effects including interstitial nephritis and hemolytic anemia. The combination of both side effects is extremely rare. In this report, we describe a case of ciprofloxacin-induced interstitial nephritis and autoimmune hemolytic anemia. Hemolytic anemia improved after stopping the drug and initiation of steroid therapy. Unfortunately, acute interstitial nephritis was irreversible and the patient developed end-stage renal disease. PMID- 12911172 TI - Hemolytic uremic syndrome: a case report from Africa. AB - Hemolytic uremic syndrome (HUS) is not a rare disease. In the tropics with high incidence of diarrhea diseases and their subsequent complications HUS can easily be missed or overlooked. We present a case of HUS highlighting its multi systemic presentation and its management in a resource poor setting where a acute dialysis and plasma exchange are limited in availability and cost. PMID- 12911171 TI - Successful treatment of renal abscess with percutaneous needle aspiration in a diabetic patient with end stage renal disease undergoing hemodialysis. AB - Renal abscesses in patients with end stage renal disease are quite rare, and misdiagnosis or delaying in diagnosis is frequent. This report examines a case of renal abscess in a patient with end stage renal disease on maintenance hemodialysis and diabetes mellitus, which presented with a prolonged fever. An infected diabetic foot was impressed initially. Purulent urine, pyuria, bacteriuria, and bacteremia were noted after admission. Renal abscess was diagnosed by percutaneous needle aspiration under computerized tomography guidance. The patient was treated with parenteral antibiotics and percutaneous aspiration of the abscess. Follow-up ultrasonography showed renal abscess resolution. This case demonstrated that nephrectomy was not required in selected uremic patients with renal abscess. PMID- 12911173 TI - Successful management of acute renal artery thromboembolism by intra-arterial thrombolytic therapy with recombinant tissue plasminogen activator. AB - Acute renal artery thromboembolism (ARAT), a rare event in native kidneys, potentially result in severe renal injury if it is not appropriately managed. The optimal therapy still remains controversial today even though various methods of managements for ARTA were applied in these decades including surgical intervention and medical approach such as thrombolytic therapy. Recombinant tissue plasminogen activator (rt-PA) reveals a better bioavailability and lower incidence of adverse effects and it has been widely used to treat a number of clinical conditions but only very few cases have been reported where rt-PA was used to treat ARAT. We described a case of ARAT, which was successfully treated by the administration of rt-PA via intra-arterial infusion within a period of 60 min without residual renal impairment. It may be a useful choice for ARAT and renal infarction sufferers. PMID- 12911174 TI - Renal and systemic amyloidosis in systemic lupus erythematosus. AB - A young male presented with oral ulceration for two years; swelling face and feet of seven days duration; diffuse goiter without signs of thyroid disease; normocytic normochromic anemia, thrombocytopenia, deranged renal functions, albuminuria of 2.5 g/24h with active urinary sediment. ANA and anti-ds DNA were positive, sonography of abdomen suggested medical renal disease. Testing for HIV, HBV, VDRL, CRP, rheumatoid factor, p-ANCA and c-ANCA were negative. Thyroid hormone assays were normal. Kidney biopsy done to stage lupus nephritis did not show any evidence of lupus involvement but staining for SAA amyloid was positive. Subsequent biopsies from the liver and rectum also stained positive for amyloid. Diagnosis of "Systemic lupus erythematosus with renal and systemic secondary amyloidosis with euthyroid diffuse goiter" was made. The case is being reported and discussed because of the interesting and rare association between amyloidosis and systemic lupus erythematosus. PMID- 12911175 TI - Permanent improvement of renal dysfunction and proteinuria with colchicine in a patient with tumoral amyloidosis and basal cell carcinoma. AB - Basal cell carcinoma and amyloid deposition was reported not uncommonly, but renal failure and nephrotic syndrome were rare. Here in, we reported a 72 years old male patient with nephrotic syndrome, renal failure and relatively small basal cell carcinoma. In this patient toumoral excision caused temporary improvement. When the proteinuria reappeared, due to amyloid deposition in toumoral tissue, colchicine was began. With colchicine therapy permanent improvement of nephrotic syndrome was achieved in follow-up 18 months. PMID- 12911176 TI - May is osteoporosis month. PMID- 12911177 TI - Anesthetic management of a patient with methemoglobinemia. PMID- 12911178 TI - Pulmonary function tests, respiratory muscle strength, and endurance of patients with osteoporosis. AB - BACKGROUND: It has been shown that patients with thoracic kyphosis due to osteoporosis have diminished pulmonary function. The aim of this study was to determine the pulmonary function, respiratory muscle strength, and endurance of patients with osteoporosis who did not have compression fractures. METHODS: The patient group consisted of 88 recently diagnosed postmenopausal osteoporotic women without spinal fractures. They were matched for age and body mass index with 54 healthy women, who formed the control group. Bone mineral density, pulmonary function test (PFT), maximum voluntary ventilation (MVV), maximal inspiratory pressure (MIP), and maximal expiratory pressure (MEP) measurements of both groups were performed. RESULTS: There were no significant differences between the two groups regarding PFT parameters and MIP and MEP. However, osteoporotic patients had significantly lower MW values. CONCLUSION: Women with postmenopausal osteoporosis without spinal compression fractures have normal PFT, MIP, and MEP values, but they have reduced respiratory muscle endurance. PMID- 12911179 TI - Attitudes of Tennessee physicians toward euthanasia and assisted death. AB - BACKGROUND: Although many studies of euthanasia and physician-assisted death (PAD) have been performed in the United States, none have specifically addressed attitudes among physicians practicing in Tennessee. METHODS: In January 2001, we mailed a 30-item survey instrument to a stratified random sample of 1,117 physicians drawn from the Tennessee Licensing Bureau. RESULTS: Tennessee physicians are highly polarized over the issues of euthanasia and assisted death. A slight majority (47%) did not favor euthanasia or PAD and would oppose the legalization of such procedures. Of the physicians supporting euthanasia or PAD (43%), only 25% would administer a lethal overdose and less than a third would counsel/prescribe medication for an overdose. Attitudes were influenced by three primary factors: ethics, religion, and the role of the physician to relieve pain and suffering. CONCLUSION: Regardless of their overall position, the majority of physicians agreed on basic restrictions and safeguards to prevent abuses and to protect vulnerable patients. PMID- 12911180 TI - Warmed versus room temperature saline for laceration irrigation: a randomized clinical trial. AB - BACKGROUND: Patient comfort is an important part of laceration repair. METHODS: The study was a randomized single-blind cross-over trial in which each participant received 250 ml warmed and 250 ml room temperature saline irrigation in simple linear wounds after a local anesthetic was instilled. The solutions and the order of irrigation (warmed first versus second) were separately randomized with a washout period between. Investigators obtaining scores were blinded. Participants determined preferred solution, whether the solution was soothing, and which caused the most discomfort. Participants rated the pain of irrigation using separate Visual Analog Scale scores. RESULTS: Thirty-eight patients were enrolled in the study. Significantly more preferred warmed to room temperature solutions (difference, 34%; 95% confidence interval, 5.7-63). A Wilcoxon signed rank test for paired data showed no order effect (P = 0.49) or difference in pain measured according to the Visual Analog Scale (P = 0.082). CONCLUSION: Warmed saline was more comfortable and soothing than room temperature saline as a wound irrigant among patients with linear lacerations. PMID- 12911181 TI - Comparison of two forms of albuterol for treatment of acute bronchospasm in pediatric patients. AB - BACKGROUND: The Circulaire nebulizer (C-Neb) is marketed as superior to low-dose albuterol for acute bronchospasm. METHODS: This clinical trial design compared clinical and fiscal outcomes of patients treated with C-Neb and high-dose albuterol (HDA). Eligibility criteria included reactive airway disease, age >5 years, and asthma score (AS) of 2 to 10. Patients with odd medical record numbers received HDA, and those with even numbers received C-Neb. Treatment was concluded if AS was 0, peak flow was >70% predicted, or after three treatments. RESULTS: One hundred eight patients were enrolled: 48 received HDA, 52 received C-Neb, and 8 were excluded. The two groups were comparable by admission rates (G2 = 0.90, P = 0.34), return visit rates (G2 = 0.73, P = 0.39), and final AS (z = 0.59, P = 0.55). Side effects were similar: change in heart rate (t = 1.61, P = 0.11) and mean arterial pressure (t = 1.36, P = 0.17). Charges per treatment, per delivery, and total charges were greater for HDA. CONCLUSION: The C-Neb is equally effective, has similar side effects, and requires less time and charges than HDA. PMID- 12911182 TI - Racial disparities in osteoporosis prevention in a managed care population. AB - BACKGROUND: Osteoporosis in black women may result in increased disability, longer hospital stays, and higher mortality compared with white women. However, it is unknown whether osteoporosis treatment or bone mineral density (BMD) measurement is different in these women, particularly in those at highest risk. METHODS: To examine differences and determinants of osteoporosis preventive interventions among white and black women in a large regional health maintenance organization, women 50 years of age and older were surveyed (n = 8,909) to determine their receipt of BMD testing and medical therapies for osteoporosis prevention. RESULTS: After adjusting for potential confounders, black women had two- to threefold lower odds of BMD test or osteoporosis prescription treatment. Even among women with a previous fracture, blacks still had a significantly lower likelihood of both BMD testing and prescription therapy. CONCLUSION: Compared with whites, black women reported significantly less BMD testing and prescription and nonprescription osteoporosis therapy. This disparity was not fully explained by other demographic or risk factor differences. PMID- 12911183 TI - Absolute versus relative values: effects on family practitioners and psychiatrists. AB - BACKGROUND: Research demonstrates that religious values clearly impact on the judgments made by physicians and their patients. One basic dichotomy--belief in ethical values that do not change (absolute values) versus belief in values that change depending on the situation (relative values)--has recently been experimentally associated with different ethical decision-making patterns. METHODS: An anonymous, randomized, national survey and 1-week response prompt with 1-month follow-up mailing as necessary were distributed to nationwide samples of board-certified American family practitioners and psychiatrists. Physicians answered descriptive questions and standardized personality assessments and responded to three vignettes describing ethically sensitive scenarios concerning birth control medication for sexually active single women, euthanasia, and abortion. RESULTS: Response rates were 34% for psychiatrists and 38% for family practitioners. Family practitioners and absolutists were significantly more supportive of religious activities and had more religious parents than psychiatrists and relativists. Furthermore, family practitioners and absolutists were less approving of the vignettes than other groups. CONCLUSION: Family practitioners were more supportive of religious activities than psychiatrists as reported in previous research. The absolute versus relative value dichotomy is a useful concept in examining physician attitudes as they affect health care and personal behavior. However, questions concerning place of worship attendance and giving in addition to specific religious value labels may be more efficient experimentally. Physicians should be aware of their own biases in discussions with patients, families, and other health care providers. PMID- 12911184 TI - Determinants of osteoporosis prevention in low-income Mexican-American women. AB - BACKGROUND: Prevention is the most cost-effective means of managing osteoporosis. However, little is known about osteoporosis-related preventive practices in Mexican-American women. We examined factors that might influence women's decision to start preventive measures for osteoporosis. METHODS: Information was gathered through a cross-sectional survey of low-income Mexican-American women who were seen at two clinics in southern Texas. RESULTS: Of the 270 participants, 37% reported calcium supplementation, and 41% reported regular weight-bearing exercise to prevent osteoporosis. Fifty (41%) of the postmenopausal women were currently using hormone replacement therapy. Only 15% of the premenopausal and 13% of the postmenopausal women recalled that their health care provider had counseled them about prevention. Multivariate analysis showed that public education, bone densitometry evaluation, knowledge of osteoporosis, and counseling were determinants of prevention. CONCLUSION: Although osteoporosis is a preventable condition, our findings suggest that the majority of Mexican American women do not receive adequate preventive measures or counseling about osteoporosis. Furthermore, we found that their health care provider's counseling about osteoporosis was a major determinant of osteoporosis prevention in these women. PMID- 12911185 TI - Surgical cricothyroidotomy in trauma patients. AB - BACKGROUND: Surgical airway intervention is occasionally necessary due to contraindicated or failed endotracheal intubation. In cricothyroidotomy patients, a choice exists between continued long-term ventilation via the cricothyroidotomy portal or conversion to tracheostomy. We examined whether conversion to tracheostomy reduces the risk of acute complications. METHODS: We retrospectively identified 46 patients with cricothyroidotomies performed at our level I trauma center over a 63-month period. We reviewed the success rate, indications, etiology, and complications. RESULTS: An airway was obtained in all cases. The most common indicator for surgical airway intervention was unsuccessful endotracheal intubation. The cause of death among nonsurvivors was not due to airway complications. Of the 15 surviving patients, 8 had conversions to tracheostomy and 7 patients did not have conversions but had decannulations. The converted group had a greater percentage of acute complications than the nonconverted group. CONCLUSION: Rate of acute complications with prolonged ventilation via cricothyroidotomy portal is equal to, if not lower than, via converted tracheostomy. Cricothyroidotomy in trauma patients may be used long term without any increase in acute complications. PMID- 12911187 TI - Analysis of 1-year vertebral fracture risk reduction data in treatments for osteoporosis. AB - One-year vertebral fracture risk reduction from clinical trials in adults with postmenopausal or glucocorticoid-induced osteoporosis is reviewed. Data were obtained by conducting a literature search of osteoporosis medications using the MEDLINE database, bibliographies of selected citations, and recent meeting abstracts. The methodologic quality of the trials was assessed using recently published criteria for ranking evidence. In prospective analyses, the 1-year risk of new morphometric vertebral fractures was reduced by risedronate 5 mg/d in two 3-year studies in postmenopausal women with prevalent vertebral fracture, and in two 1-year studies in patients with or at risk for glucocorticoid-induced osteoporosis. The 1-year risk of clinical vertebral fractures was reduced by alendronate and raloxifene in post hoc analyses. Reduction of morphometrically identified vertebral fracture risk is a more stringent therapeutic goal than clinical vertebral fracture risk. Therefore, more weight should be given to data from studies that use the morphometry to assess vertebral fracture incidence. PMID- 12911186 TI - Selective estrogen receptor modulators. AB - Because of recent concerns about the long-term risks of estrogen replacement therapy in postmenopausal women, there is growing interest in a group of compounds known as selective estrogen receptor modulators (SERMs). The SERMs bind to estrogen receptors and have tissue-specific effects that allow them to function as estrogen agonists in some tissues and estrogen antagonists in other tissues. There are four SERMs currently marketed in the United States. These include the triphenylethylenes--clomiphene citrate (Clomid), tamoxifen, and toremifene--and the benzothiophene, raloxifene. Clomid is used primarily in the treatment of infertility. Tamoxifen is indicated for the treatment and prevention of breast cancer. It has an estrogen antagonist effect on breast tissue, but an estrogen-like effect on lipids, bone, and the endometrium. Toremifene has an antagonist/agonist profile similar to that of tamoxifen. Raloxifene is approved for the prevention of osteoporosis in postmenopausal women. It is thought to be an estrogen antagonist on the uterus and breast tissues and an estrogen agonist with respect to bone and serum lipids. PMID- 12911188 TI - Brown recluse spiders: a review to help guide physicians in nonendemic areas. AB - The brown recluse spider (Loxosceles reclusa) exists in significant populations mainly in the midwestern United States. Although bites can cause significant harm, envenomation is infrequent, and cases are usually clinically insignificant. Proper knowledge of the spider's habitat and lifestyle as well as the signs and symptoms of loxoscelism are needed to assess clinical cases adequately. Loxoscelism can masquerade as many serious pathologies, and vice versa, so it is important for the clinician to explore all possibilities in the differential diagnosis thoroughly. Treatments are controversial, and no conclusive test for envenomation is currently available. This review provides information to help physicians, especially in nonendemic areas, include or exclude brown recluse bite in the clinical diagnosis and provide proper care. PMID- 12911189 TI - Bone marrow amyloidosis with erythropoietin-resistant anemia in a patient undergoing chronic hemodialysis treatment. AB - The resistance to erythropoietin, which is used to treat normochromic, normocytic anemia in chronic renal failure, can develop in patients with conditions such as iron deficiency, aluminum toxicity, hyperparathyroidism, chronic inflammatory diseases, and primary hematological disorders. We found amyloidosis in the bone marrow of a woman without any other etiology for erythropoietin resistance who was undergoing chronic hemodialysis. Her anemia did not improve, despite 6 months of erythropoietin therapy. Bone marrow amyloidosis was found to be the reason for erythropoietin-resistant anemia in our patient with chronic renal failure and renal anemia. We present the case of bone marrow amyloidosis because it is a very rare cause of erythropoietin resistance. PMID- 12911190 TI - Intracerebral tuberculoma misdiagnosed as neurosarcoidosis. AB - We describe two patients who had tuberculomas that were initially misdiagnosed as neurosarcoidosis, leading to prolonged steroid therapy before initiation of antituberculous medications. Neither patient was infected with the human immunodeficiency virus, and one of the patients had a negative tuberculosis skin test. PMID- 12911191 TI - Acute renal infarction as a cause of low-back pain. AB - Detection of acute renal infarction is often delayed or missed because of both the rarity of the disease and its nonspecific clinical presentation. Abrupt onset of low-back pain in a patient at high risk for a thromboembolic event may be the first indication of renal infarction. We report a case of acute renal infarction and review its diagnosis and management. PMID- 12911192 TI - Acute metabolic decompensation in an adult patient with isovaleric acidemia. AB - Isovaleric acidemia is a rare autosomal recessive inborn error of leucine catabolism caused by deficiency of isovaleryl coenzyme A dehydrogenase. This enzymatic deficiency leads to severe metabolic derangement, manifested clinically as vomiting, dehydration, and acidosis progressing to seizures, coma, and death. The two phenotypic expressions are the acute severe and the chronic intermittent form. The acute severe phenotype typically results in death during early infancy, whereas patients with the chronic intermittent form are asymptomatic at baseline but have episodes of acute metabolic decompensation, usually in the setting of infection, physical exertion, or ingestion of protein-rich food. This case illustrates how inborn errors of metabolism resulting in organic acidemia can be manifested in adults and why the internist needs to be aware of them. PMID- 12911193 TI - Anesthetic management of a patient with methemoglobinemia. AB - Methemoglobinemia results from the oxidation of the ferrous iron in hemoglobin to the ferric iron state. Methemoglobin is incapable of carrying O2, and high levels may impact on O2 delivery to the tissues. Methemoglobinemia may result from congenital deficiencies of enzymes that normally convert methemoglobin to hemoglobin, alterations in the hemoglobin molecule itself or, most commonly, from the ingestion of medications or toxins that oxidize the ferrous iron of hemoglobin. Several issues must be considered when anesthetizing patients with methemoglobinemia, including the potential for decreased O2 delivery, which may be exacerbated by intraoperative blood loss and anemia, interference with normal intraoperative monitoring devices, and the potential for medications to cause or exacerbate methemoglobinemia. We describe a patient with acquired methemoglobinemia from dapsone therapy who required anesthetic care for shoulder arthroscopy. The patient's drug-induced methemoglobinemia was diagnosed intraoperatively during previous anesthesia on the basis of discrepancy between the O2 saturation noted by pulse oximetry and that obtained from arterial blood gas analysis. Anesthetic care for patients with methemoglobinemia is discussed and a review of methemoglobinemia presented. PMID- 12911194 TI - Pleural effusion caused by trapped lung. AB - We report a case of pleural effusion from trapped lung secondary to sarcoidosis. The patient presented with dyspnea, right pleural effusion, left and right upper lobe infiltrate and a widened mediastinum. The pleural effusion and dyspnea failed to respond to a course of oral corticosteroids but was relieved by decortication with stripping of inflammatory fibrous bands encasing the visceral pleural. Histologic examination of the lung, visceral and parietal pleura, and the fibrous adhesions all revealed noncaseating granulomas. PMID- 12911195 TI - Malignant gastric stromal tumor: unusual metastatic patterns. AB - We present the case of a woman with metastatic malignant gastric stromal tumor occurring four years following a partial gastrectomy and distal esophagectomy, which presented as atypical breast mass with synchronous occult gastrointestinal bleeding. A discussion of metastatic pattern is presented, with emphasis placed on the need for continued surveillance after resectional surgery for gastrointestinal stromal tumor (GIST). PMID- 12911196 TI - Esophageal perforation and mediastinitis from fish bone ingestion. AB - Esophageal perforation is a serious condition with a high mortality rate. Successful therapy depends on the size of the rupture, the time elapsed between rupture and diagnosis, and the underlying health of the patient. Common causes of esophageal perforation include medical instrumentation, foreign-body ingestion, and trauma. A case of esophageal perforation due to fish bone ingestion in a 70 year-old diabetic male is described here, with a review of the pertinent literature. The patient presented with odynophagia after a meal that included fish. Initial evaluation was nondiagnostic and the patient was discharged home. The patient returned 12 days later with fever, generalized weakness, and persistent dysphagia. Esophageal biopsy of a necrotic ulcer revealed foreign material with acute inflammatory changes. Computed tomography scan demonstrated a pneumomediastinum. The patient became hemodynamically unstable and died on the third hospital day. PMID- 12911197 TI - Radiation pneumonitis successfully treated with inhaled corticosteroids. PMID- 12911198 TI - Spontaneous pleural empyema due to Yersinia enterocolitica. AB - Yersinia enterocolitica is a well-known cause of enterocolitis. Although focal extraintestinal manifestations and disseminated disease have been described, usually in immunosuppressed patients, infection in the chest seems to be rare. We report the case of an alcoholic man who had spontaneous pleural empyema due to Y. enterocolitica. PMID- 12911199 TI - Persistent left superior vena cava: an intensivist's experience and review of the literature. AB - Persistent left superior vena cava is a relatively rare vascular anomaly. It is, however, the most common variation in the thoracic venous system. The anomaly is typically detected on chest x-ray after the placement of a pulmonary artery catheter or pacemaker leads. This report describes a case of persistent left superior vena cava detected after successful placement of a pulmonary artery catheter. PMID- 12911201 TI - Comparison of adjuvant anesthetics for propofol induction. AB - PURPOSE: Fentanyl was compared with nitrous oxide/sevoflurane as an adjuvant anesthesia to propofol during induction. METHODS: Two-hundred sixty-three patients of American Society of Anesthesiologists physical status 1 or 2 undergoing minor surgery were randomly divided into two groups. Group F patients (n = 125) received 2 microg x kg(-1) fentanyl and 1.8 mg x kg(-1) propofol, and were ventilated by mask with oxygen. Group S patients (n = 138) received 1.8 mg x kg(-1) propofol, followed by inhalation of 4% sevoflurane in N2O (41 x min(-1)) and oxygen (21 min(-1)) by mask. The trachea was intubated exactly 2, 3, 4, or 5 min after injection of 0.1 mg x kg(-1) vecuronium, and the conditions of endotracheal intubation were scored according to the patients' responses to laryngoscopy and endotracheal intubation. Systolic blood pressure (SBP) and heart rate (HR) were measured before and after endotracheal intubation. The cost of anesthetics was also calculated. RESULTS: No significant differences in SBP were observed between the groups throughout the induction period. HR did not change from preanesthetic values in group F. In contrast, HR in group S patients increased by 9-18 beats x min(-1) (bpm) after inhalation of N2O/sevoflurane and further increased by 17-21 bpm following endotracheal intubation. Significant differences in HR were noticed between the groups (P < 0.001). The conditions of endotracheal intubation were similar in the two groups and were satisfactory when mask ventilation exceeded 3 min. Fentanyl was less expensive than sevoflurane/N2O anesthesia when mask ventilation exceeded 3 min. CONCLUSION: From the standpoints of hemodynamics and drug cost, fentanyl is preferable to N2O/sevoflurane inhalation as an adjuvant to propofol during induction, because mask ventilation for more than 3 min was required for satisfactory endotracheal intubation. PMID- 12911200 TI - Chronic aortic dissection as a cause of fever of unknown origin. PMID- 12911202 TI - Hemodynamic and catecholamine responses during tracheal intubation using a lightwand device (Trachlight) in elderly patients with hypertension. AB - PURPOSE: Tracheal intubation using a lightwand device (Trachlight) should minimize hemodynamic change by avoiding direct-vision laryngoscopy. We evaluated hemodynamic and catecholamine responses during tracheal intubation using a Trachlight in elderly patients with hypertension. METHODS: Twenty-six hypertensive patients aged over 65 years undergoing orthopedic surgery were randomly divided into two groups, group L (n = 13) and group T (n = 13). Anesthesia was induced with fentanyl (2 microg x kg(-1)) and propofol (1.5 mg x kg(-1)), and then muscle relaxation was obtained with vecuronium (0.15 mg x kg( 1)). The trachea was intubated with either a Macintosh laryngoscope (group L) or a Trachlight (group T). Hemodynamics, plasma catecholamine concentrations, and arterial blood gases were measured before the induction of anesthesia (T0), before tracheal intubation (T1), immediately after tracheal intubation (T2), and 3 min after tracheal intubation (T3). RESULTS: The intubation time was shorter in group T than in group L (12.6 +/- 1.7 vs 23.5 +/- 2.9 s, mean +/- SE; P < 0.01). Compared with the preinduction (T0) value, systolic blood pressure (SBP) showed a significant decrease at T1 and T3 in group L and at T1, T2, and T3 in group T. The heart rate (HR) and plasma norepinephrine (NE) concentration showed no change in either group throughout the time course, whereas the plasma epinephrine (E) concentration showed a significant decrease at T2 and T3 in both groups. The mean values of the rate-pressure product (RPP: HR x SBP) were less than 15000 after tracheal intubation in both groups. There was no significant difference in hemodynamic or catecholamine responses between groups at any point. No patient had ischemic ST-T changes in either group. CONCLUSION: A lightwand has no advantage over a laryngoscope in terms of hemodynamic and plasma catecholamine responses to tracheal intubation in elderly patients with hypertension, despite a shorter intubation time. PMID- 12911203 TI - Temperature and humidity of the Drager Cato anesthetic machine circuit. AB - PURPOSE: The Drager Cato anesthetic machine (Drager, Lubeck, Germany) effectively humidifies and warms anesthetic gases, because it has a built-in hotplate to heat the breathing system, and expired gas passes through the CO2 absorbent three times during one breath. In the present study, we measured the temperature and absolute humidity (AH) of the anesthetic circuit in the Drager Cato machine with and without heat moisture exchangers (HME), and compared them with those in another anesthetic machine, the Aestiva/5 (Datex-Ohmeda, Helsinki, Finland). METHODS: Forty-eight adult patients were randomly assigned to one of eight groups according to the anesthetic machine, fresh gas flow (FGF), and the use of HME (n = 6 each): Cato 0.51 x min(-1) without HME (group 1), Cato 1.01 x min(-1) without HME (group 2), Cato 0.51 x min(-1) with HME (group 3), Cato 1.01 x min(-1) with HME (group 4), Aestiva 0.51 x min(-1) without HME (group 5), Aestiva 1.01 x min( 1) without HME (group 6), Aestiva 0.51 x min(-1) with HME (group 7), and Aestiva 1.01 x min(-1) with HME (group 8). The temperature and AH of the anesthetic gases were measured with a Moiscope (S.K.I. Net, Tokyo, Japan), which was placed between the endotracheal tube and the Y-piece of the anesthetic circuit. The HME was placed between the Moiscope and the Y-piece of the anesthetic circuit. The temperature and AH of the anesthetic gases were measured at 5, 10, and 15 min and then every 15 min up to 150 min after tracheal intubation. RESULTS: Among the groups without HME (groups 1, 2, 5, and 6), the inspired temperatures and AH in groups 1 and 2 were significantly higher than those in groups 5 and 6 at all times during the study period (P < 0.01-0.001). The inspired temperatures and AH of the groups with HME (groups 3, 4, 7, and 8), were significantly higher than those in groups 2, 5, and 6 (P < 0.01-0.001). Among the groups with HME, the AH in group 3 was significantly higher than that in group 8 until the final study period. CONCLUSION: The present study indicates that the Drager Cato machine was more effective in warming and humidifying respiratory gas than the Aestiva/5, and that Aestiva/5 without HME does not reach the optimal temperature and humidity ranges, even if minimal flow anesthesia (0.51 x min(-1)) is performed. PMID- 12911205 TI - Anesthetic implications of neuromuscular disease. AB - Neuromuscular disease covers a wide range of conditions, with anesthesia management being required either for problems relevant to the disorder or for comorbid conditions. The diseases often have specific problems that can usually be predicted from their pathophysiology. The anesthesiologist must ensure a thorough preoperative assessment, appropriate choice of anesthetic technique and neuromuscular blocking drugs, and careful monitoring of both hemodynamic parameters and the degree of neuromuscular blockade. With these considerations, the patient with neuromuscular disease, although challenging, can be given anesthetic care in a safe fashion. PMID- 12911204 TI - Comparison of early postoperative quality of life in minimally invasive versus conventional valve surgery. AB - PURPOSE: Minimally invasive cardiac surgery (MICS), an approach in which full sternotomy is avoided and the surgical incision is minimal, has been shown to produce less postoperative discomfort and to enable earlier mobilization and discharge than conventional cardiac surgery (CCS). This study was performed to retrospectively evaluate quality of life following MICS in comparison with CCS valve surgery. METHODS: Sixty-six patients scheduled for MICS and 50 patients scheduled for CCS for isolated aortic or mitral valve surgery from January 1999 to June 2001 were enrolled in the study. The clinical records for the two groups were compared across intraoperative parameters and those associated with postoperative quality of life. RESULTS: The aortic clamp and cardiopulmonary bypass times in the MICS group were longer than those in the CCS group (144 +/- 42 and 224 +/- 58min vs 112 +/- 21 and 179 +/- 27min, P < 0.001). Postoperative pain medication (rectal buprenorphine and intramuscular pethidine) was administered to 18 of the 66 MICS patients (27%), as compared with 26 of the 50 CCS patients (52%, P = 0.007). Postoperative delirium was less frequent in the MICS group than the CCS group (26% vs 44%, P = 0.039). Initial postoperative food intake and urine catheter removal were possible earlier in the MICS than in the CCS group. MICS patients had shorter stays in the intensive care unit than CCS patients (37.4 +/- 7.3 vs 45.9 +/- 8.7h, P < 0.001). CONCLUSION: Although longer aortic clamp and cardiopulmonary bypass times remain a problem in MICS procedures, our results suggest that MICS, as compared with CCS, facilitates earlier recovery of daily activities and provides improved quality of life in the early postoperative period. PMID- 12911206 TI - Rapid improvement of respiratory symptoms associated with fat embolism by high dose methylpredonisolone: a case report. PMID- 12911207 TI - Satisfactory recovery of consciousness after prolonged cardiopulmonary resuscitation due to unexpected hemorrhage during a conduit ileum examination. PMID- 12911208 TI - Anesthetic management of a patient with tuberous sclerosis presenting for renal transplantation. PMID- 12911210 TI - Severe anaphylactoid reaction to ranitidine in a parturient with subsequent fetal distress. PMID- 12911209 TI - Silent gastric perforation in a pancreatic cancer patient treated with neurolytic celiac plexus block. PMID- 12911211 TI - Pressure sore as a possible complication of lower central neuraxial blockade. PMID- 12911212 TI - Hemodynamic effects of felypressin and epinephrine on anesthetized rats. PMID- 12911214 TI - Effects of intermittent pneumatic foot compression on blood coagulability and fibrinolysis assessed by a whole blood viscometer Sonoclot. PMID- 12911213 TI - Absence of efficacy of ultrasonic two-way Doppler flow detector in routine percutaneous arterial cannulation. PMID- 12911215 TI - Corneal abrasion after the wake-up test in spinal surgery. PMID- 12911216 TI - Unusual failure of intubation. PMID- 12911217 TI - Editorial: Professor Silvia Braslavsky. PMID- 12911218 TI - Time-resolved photothermal methods: accessing time-resolved thermodynamics of photoinduced processes in chemistry and biology. AB - Photothermal methods are currently being employed in a variety of research areas, ranging from materials science to environmental monitoring. Despite the common term which they are collected under, the implementations of these techniques are as diverse as the fields of application. In this review, we concentrate on the recent applications of time-resolved methods in photochemistry and photobiology. PMID- 12911219 TI - Structural changes upon excitation of D1-D2-Cyt b559 photosystem II reaction centers depend on the beta-carotene content. AB - Different preparations of D1-D2-Cyt b559 complexes from spinach with different beta-carotene (Car) content [on average from <0.5 to 2 per reaction center (RC)] were studied by means of laser-induced optoacoustic spectroscopy. phiP680(+)Pheo( ) does not depend on the preparation (or on the Car content) inasmuch as the magnitude of the prompt heat (produced within 20 ns) does not vary for the different samples upon excitation at 675 and 620 nm. The energy level of the primary charge-separated state, P680(+)Pheo(-), was determined as EP680(+)Pheo(-) = 1.55 eV. Thus, an enthalpy change accompanying charge separation from excited P680 of deltaH*P680Pheo-->P680(+)Pheo(-) = -0.27 eV is obtained. Calculations using the heat evolved during the time-resolved decay of P680(+)Pheo(-) (< or = 100 ns) affords a triplet (3[P680Pheo]) quantum yield phi3[P680Pheo] = 0.5 +/- 0.14. The structural volume change, deltaV1, corresponding to the formation of P680(+)Pheo(-), strongly depends on the Car content; it is ca. -2.5 A3 molecule( 1) for samples with <0.5 Car on average, decreases (in absolute value) to -0.5 +/ 0.2 A3 for samples with an average of 1 Car, and remains the same for samples with two Cars per RC. This suggests that the Car molecules induce changes in the ground-state RC conformation, an idea which was confirmed by preferential excitation of Car with blue light, which produced different carotene triplet lifetimes in samples with 2 Car compared to those containing less carotene. We conclude that the two beta-carotenes are not structurally equivalent. Upon blue light excitation (480 nm, preferential carotene absorption) the fraction of energy stored is ca. 60% for the 9Chl-2Car sample, whereas it is 40% for the preparations with one or less Cars on average, indicating different paths of energy distribution after Car excitation in these RCs with remaining chlorophyll antennae. PMID- 12911220 TI - Protein relaxation in the photodissociation of myoglobin-CO complexes. AB - Laser-induced optoacoustic spectroscopy has been applied to the study of the photodissociation of myoglobin-CO complexes. Time-resolved optoacoustic signals have been measured from aqueous solutions of horse myoglobin-CO complex (hMbCO) at pH 3.5 and 8, and of sperm whale myoglobin-CO complex (swMbCO) at pH 8, in the temperature range 273-300 K. The signal of hMbCO at pH 8 exhibits three components. The first, which is faster than 20 ns and is associated with a reaction enthalpy of 61 kJ mol(-1), corresponds to Fe-CO bond breakage. The second component has a decay time of 80 ns at 293 K and is associated with an exothermic protein relaxation (-13 kJ mol(-1)) and a volume change of -3 ml mol( 1). The relaxation, which involves a state where the photo-dissociated CO is still in a protein docking site, is thermally activated, with an activation enthalpy of 51 kJ mol(-1). The third component has a decay time of 800 ns at 293 K and an activation enthalpy of 39 kJ mol(-1), and is associated with an endothermic process (26 kJ mol(-1)) and an expansion of 19 ml mol(-1). This process is ascribed to the migration of the photodissociated CO to the bulk solvent. At acidic pH, the latter process becomes faster (230 ns) and the volume change decreases. These features are correlated with the presence of an open form of the protein. swMbCO exhibits two components only, due to the overlap of the two fastest processes. The first involves a reaction enthalpy of 49 kJ mol(-1) and a volume contraction of -4.9 ml mol(-1). The second component (900 ns at 293 K, activation enthalpy 45 kJ mol(-1)) is associated with a reaction enthalpy of 38 kJ mol(-1) and a volume expansion of 15.3 ml mol(-1). These experimental findings have been interpreted by means of a new model, which also takes into account both laser flash photolysis results and structural information. The model is based on a two-dimensional scheme which describes both protein relaxation and the CO pathway following photodissociation. PMID- 12911222 TI - Two-photon photoacoustic calorimetry and the absolute measurement of molar absorption coefficients of transient species in solution. AB - The theory of two-photon photoacoustic calorimetry (PAC) is developed for the case of a homogeneously irradiated volume and it is shown that the laser intensity dependence of the photoacoustic signals can be used to determine the molar absorption coefficient of transient species. The application of the method is illustrated via the measurement of the absorption coefficients of benzophenone and acetophenone triplets using a front-face PAC cell. The very high sensitivity exhibited by this cell strongly recommends the inclusion of laser-intensity dependence studies in the procedure for measuring heat depositions. Only extrapolation to zero laser intensity can afford reliable enthalpies of formation for very short-lived species. PMID- 12911221 TI - Photochromic chromopeptides derived from phycoerythrocyanin: biophysical and biochemical characterization. AB - Truncated chromopeptides have been prepared from the small photo- and redox switchable biliprotein alpha-phycoerythrocyanin (alpha-PEC). The native chromoprotein consists of a C-terminal globin domain containing the chromophore and the regulatory cysteins 98 and 99, and a two-helix (X,Y) N-terminal domain responsible for aggregation. Digestion with chymotrypsin-free trypsin leads to three chromopeptides, (N-30, N-33 and N-35), basically lacking the two N-terminal helices X and Y. The photo- and redox chemistry of the major product (N-33) is identical, qualitatively and quantitatively, to that of native alpha-PEC. A series of N- and C-terminally truncated polypeptides were expressed in E. coli and subjected to autocatalytic and enzymatic reconstitution with phycocyanobilin. Enzymatic reconstitution was possible with N-terminally truncated polypeptides up to 45 aa, while neither a more extensively shortened (N-63) peptide, nor two C terminally shortened polypeptides could be reconstituted. All chromopeptides recovered from enzymatic reconstitution contained the native phycoviolobilin chromophore and showed the photochemical and redox reactivity of alpha-PEC, albeit quantitatively reduced in the N-45 chromopeptide. PMID- 12911223 TI - Application of photoacoustic calorimetry to the determination of volume changes in reactions involving radical anions in aqueous solutions. AB - The particular features of the application of photoacoustic calorimetry to the determination of volume changes in reactions involving radical anions in aqueous solutions are addressed. Analysis of recent literature data on volume changes of redox pairs, including our previous results on the couples NO2(-)/NO2 and SO4(2 )/(SO4)*-, together with known values of molal volume changes for ionization reactions, allow us to derive an empirical correlation for these categories of reactions. Finally, the pertinence of describing the volume changes of complex processes as a simple sum of separable terms is critically examined. PMID- 12911224 TI - Listening to the blue: the time-resolved thermodynamics of the bacterial blue light receptor YtvA and its isolated LOV domain. AB - YtvA is a bacterial flavo-protein related to plant phototropin. The photochemistry of YtvA and of its isolated LOV domain (YtvA-LOV) has been characterized by optical, mass spectrometric and photocalorimetric methods. The energy content (E390) of the FMN-C4a-thiol photoadduct (YtvA390 and YtvA-LOV390) and its structural volume change (deltaV390), with respect to the parent state, have been determined by means of Laser Induced Optoacoustic Spectroscopy (LIOAS). The high value of E390, 136 and 115 kJ mol(-1), respectively, ensures a large driving force for the dark recovery to the unphotolyzed state and points to a strained conformation of the protein or/and the chromophore in the photoadduct. The value of deltaV390 is significantly different for the two proteins, deltaV390 = -12.5 ml mol(-1) in YtvA and -17.2 ml mol(-1) in YtvA-LOV. The kinetics of the dark recovery reaction for YtvA-LOV is slower than for full-length YtvA, with taurec = 3900 and 2600 s at 25 degrees C, respectively, and shows a different temperature dependence. A similarly slow kinetics can be induced in YtvA by high ionic strength. Minor differences are observed in the fluorescence and photoadduct formation quantum yield. The overall stability is higher for YtvA than for YtvA-LOV. The data as a whole are indicative of an interaction between the two domains of YtvA, most probably mediated by electrostatic interactions that renders the full-length protein a compact and more rigid unit. The results reported here support the idea that the formation of the photoadduct changes the dynamics of the protein, depending on the conformational flexibility of the parent state. Flashing of the photoadduct induces a negligible deltaV, with 96% of the excitation energy dissipated as heat in <20 ns, indicating that the photoadduct does not undergo a photocycle on the LIOAS time scale, or that the photoinduced reactions occur with very low yield. PMID- 12911225 TI - Photoreaction of caged ATP studied by the time-resolved transient grating method. AB - The photoreaction of caged ATP, P3-[1-(2-nitrophenyl)ethyl]adenosine-5' triphosphate, has been investigated using the time-resolved transient grating (TG) method. We found that a feature of the TG signal time profile depends sensitively on the grating wavenumber (q) after the photoexcitation of caged ATP. This q-dependent feature of the TG signal was interpreted based on a model where the ATP release rate is comparable to the molecular diffusion process. We found that the TG signals at various q can be consistently analyzed based on this model and the ATP release rate determined. The enthalpy and volume changes of the reaction have been determined by quantitative measurement of the grating and photoacoustic signals. PMID- 12911226 TI - Monitoring of road-traffic emissions with a mobile photoacoustic system. AB - Ammonia and ethene are pollutants which are arousing concern as regards their environmental impact, e.g. as greenhouse gases. Road traffic is an increasingly important emission source for these gases. As part of an atmospheric pollution measurement campaign, we performed in situ measurements of NH3, C2H4 and CO2 concentrations at the mouth of a freeway tunnel with a time resolution of 1 min using a mobile laser-based photoacoustic system. Measurements were performed over a period of 5 weeks. In good temporal correlation with traffic counts, we observed peak concentrations of >600 ppb (>1200 ppb for Friday afternoon peaks) for NH3, of up to 400 ppb for C2H4 and >2000 ppm for CO2. Preliminary comparisons with previous measurements at the same location indicate a considerable increase in ammonia and CO2 peak concentrations (greater than the increase in traffic over the same period) and a less pronounced increase in C2H4 concentrations. PMID- 12911227 TI - Steady-state absorption rate models for use in relaxation rate studies with continuous laser excited photothermal lens spectrometry. AB - This paper examines the solutions of kinetic rate equations for prediction of the photothermal lens signals under irradiance conditions that can lead to optical saturation or bleaching. The relaxation kinetics resulting from forcing excited state populations in multiple levels by high excitation irradiance continuous lasers is examined and irradiance-dependent photothermal lens signals are predicted. The analyses described in this paper are based on simple kinetic models for optical excitation and subsequent excited state relaxation. Dark-state relaxation is assumed to be extremely fast compared to limiting kinetics resulting in simplified excited state models. Kinetic models are derived for two, four and five active level molecular systems. Gaussian laser beam profiles are assumed and time dependent photothermal lens signals are calculated. Models account for excitation laser profile, thermal relaxation of the spatially and temporally distorted heating rate distribution resulting from nonlinear absorption, and metastable state relaxation. This heating rate is used to calculate the temperature change distribution and subsequently the optical elements needed to model the experimental photothermal signals. PMID- 12911228 TI - Measurement of enthalpy and volume changes in photoinitiated reactions on the ms timescale with a novel pressure cell. AB - Time-resolved photoacoustics is an excellent method with which to measure enthalpy and volume changes of photochemical and photobiological reactions. However, it fails at times longer than approximately 10 micros. The design principles of a pressure or volume cell covering the time range of 20 micros to several seconds is presented. The sensitivity of the cell has been verified and its application to the photocycle of bacteriorhodopsin is presented. Because of the similar cell structure and data analysis it is now possible to determine enthalpy and volume changes in photo-initiated reactions over the timescale of nanoseconds to seconds with the same solution. PMID- 12911229 TI - Thermodynamic changes associated with the formation of the hydrated electron after photoionization of inorganic anions: a time-resolved photoacoustic study. AB - The enthalpy and volume changes, deltaH and deltaV, associated with the 266 nm laser-induced photoionization reactions of aqueous ferrocyanide and iodide ions, to yield the hydrated electron, e(-)aq, and oxidized products were determined by temperature-dependent time-resolved photoacoustics. The photoionization quantum yield as function of temperature (9-30 degrees C) was determined by laser flash photolysis actinometry. The obtained values were used for the calculation of thermodynamic parameters associated with the formation of e(-)aq, such as the apparent partial molar volume, V(o)e = 26 cm3 mol(-1), and the standard formation enthalpy and entropy changes, deltaH(o)f,e = 31 kJ mol(-1) and TdeltaS(o)f,e = 338 kJ mol(-1). These results indicate that the formation of the aqueous excess electron solution is governed by the increase in entropy in the three-dimensional hydrogen-bonding network of water. PMID- 12911230 TI - Deprotonation yields, pKa, and aci-nitro decay rates in some substituted o nitrobenzaldehydes. AB - In this paper we report the deprotonation yields, the pKa, and decay kinetics of the aci-nitro intermediates of some substituted 2-nitrobenzaldehydes that can be used as photoactivatable caged proton compounds. The decay of the aci-nitro absorbance for 2-nitrobenzaldehyde occurs within a few nanoseconds from photoexcitation. Addition of electron donating methoxy substituents at positions 4 and 5 leads to lower deprotonation yields, higher pKa, and slower decays of the aci-nitro intermediates. On the contrary, the decay rate is accelerated by the introduction of an electron-withdrawing Cl atom at position 4 in the phenyl ring, with little influence on the deprotonation yield and pKa of the aci-nitro intermediate. PMID- 12911231 TI - Excited state absorption of fullerenes measured by the photoacoustic calorimetry technique. AB - Photoacoustic calorimetry (PAC) is used to determine the excited state absorption cross sections in a molecular system showing reverse saturable absorption behavior. PAC experiments on fullerene and fulleropyrrolidine in toluene solutions are performed at 532 nm and 690 nm, with a ns laser source. The PAC signal amplitude displays a superlinear increase when the energy of the applied laser source is increased. This behavior is ascribed to a process of enhanced absorption due to molecules populating the excited electronic states. The PAC signal observed for these chromophores is simulated numerically. The simulations rely on a description of the absorbing molecule as a six-level system, whose molecular parameters (i.e. absorption cross sections and lifetimes) are the ones for a reverse saturable absorber. The time-dependent population in the different energy levels is described through a rate equation system. This kind of model has been widely used by us to reproduce other experimental data such as nonlinear transmittance and Z-scan data. The PAC signal amplitude is the sum of the different contributions to non-radiative relaxation which arise from molecules populating different energy levels. The absorption cross sections for the singlet and triplet excited states of fullerene and fulleropyrrolidine are derived from the simulated PAC signal amplitudes. The values obtained are in good agreement with literature data measured with different techniques. PMID- 12911232 TI - The photophysics of nitrocarbazoles studied by using spectroscopic, photoacoustic and luminescence techniques. AB - The photophysical behaviour of 1-nitrocarbazol and 3-nitrocarbazol in different organic solvents has been studied by using spectroscopic, luminescence and photoacoustic techniques. Absorption spectra and triplet state transient spectra were recorded and formation quantum yields and lifetimes determined. Phosphorescence emission at 77 K and laser induced luminescence in acetonitrile solution at 298 K for the nitrocarbazoles were studied. Results are discussed in terms of the optimized structure of 1-nitrocarbazole and 3-nitrocarbazole and compared with that of carbazole. The nitro group attached to the carbazole moiety induces important changes in the photophysical behavior of those compounds. PMID- 12911233 TI - Volume of reaction, energetics, and kinetics of tetrahydrothiophene displacement of solvent from Mo(CO)5(alkane). AB - Laser-induced optoacoustic spectroscopy (LIOAS) was used to determine the enthalpies and volumes of reactions for each step in ligand substitution of molybdenum hexacarbonyl with tetrahydrothiophene (THT). The enthalpy for substitution of CO on Mo(CO)6 by an alkane and of coordinated alkane on Mo(CO)5(alkane) by THT are 114 and -99 kJ mol(-1), respectively. Likewise, the volumes of reaction are 13 and -16 ml mol(-1). These results allow the calculation of the bond energies for Mo-alkane (56 kJ mol(-1)) and Mo-THT (155 kJ mol(-1)). Furthers studies should reveal that, for certain cases, the volume of reaction can be neglected in the determination of enthalpies of ligand substitution by LIOAS. PMID- 12911234 TI - Ultrafast absorption and photothermal studies of decarboxytrichochrome C in solution. AB - The trichrochromes are natural constituents of pheomelanins. Herein, time resolved spectroscopic techniques are used to quantify the energetics and dynamics of the primary photoprocesses of trichochromes following excitation into the lowest excited singlet state of the molecule. The absorption spectrum of decarboxytrichochrome C (dTC) reveals multiple electronic states are accessible upon visible and UV excitation. Emission is not observed upon excitation into the lowest energy absorption band. Photothermal measurements reveal 91% of the photon energy is promptly released as heat. Femtosecond time-resolved absorption studies reveal a ground-state recovery time of approximately 2.4 ps. Complete recovery of the ground state is not observed; 15% of the initially excited molecules do not recover on the approximately 150 ps timescale. The combination of ultrafast absorption and photoacoustic data suggest photoexcitation produces a long-lived intermediate and the energy of this species is at least 133 kJ mol(-1) above the ground state of the dTC molecule. PMID- 12911235 TI - Cyclooxygenase-2 inhibitors and nonsteroidal anti-inflammatory drugs in the management of arthritis. AB - The management of arthritis pain should be individualized to the needs and characteristics of the patient. The decision to use nonsteroidal anti inflammatory drugs (NSAIDs), and in particular the cyclooxygenase-2 (COX-2) inhibitors, is multidimensional. The challenge is to achieve optimal pain relief at the minimum dose to minimize adverse effects. Whenever possible, NSAIDs should be given as monotherapies or in combinations at the lowest effective doses. The COX-2 inhibitors are a safe choice for most patients who are at low risk for a cardiovascular event. Individuals who are at risk for thromboses should not receive unopposed COX-2 inhibitors; COX-2 should be given in combination with low dose aspirin which is expected to be cardioprotective in high-risk patients. PMID- 12911236 TI - Chondroprotective agents: glucosamine and chondroitin. AB - The near universal finding of the safety of glucosamine and chondroitin combined with some compelling evidence of their efficacy should spur further research into their mechanism of action, optimal dosing, long-term effects on disease modification, and clinical applicability. When recommending a supplement to patients, the clinician should take into account the purity of the ingredients, reputation of the manufacturer, and the molecular weight of chondroitin supplied. PMID- 12911237 TI - Intra-articular hyaluronan therapy. AB - Viscosupplementation, in which hyaluronan derivatives are injected into the intra articular space of osteoarthritic joints, is now widely used to treat knee osteoarthritis (OA). No viscosupplements have been approved for osteoarthritic joints other than the knee. To date, no clinical trials using viscosupplements to treat ankle or foot OA have been published. However, the mechanisms thought to be responsible for viscosupplementation's therapeutic effects would likely apply in any synovial joint. A goal of this article is to stimulate interest in research to assess the potential role of viscosupplementation in treating foot and ankle OA. PMID- 12911238 TI - Systematic review of treatment strategies for osteochondral defects of the talar dome. AB - The aim of this study was to summarize all eligible studies to compare the effectiveness of different treatment strategies for osteochondral defects (OCD) of the talus. Electronic databases from January 1966 to June 2000 were systematically screened. Based on our inclusion criteria, 39 studies describing the results of treatment strategies for OCD of the talus were included. No randomized clinical trials (RCT) were identified. Fourteen studies described the results of nonoperative treatment (NT); 4: the results of excision alone; 10: the results of excision and curettage (EC); 21: the results of excision, curettage, and drilling (ECD); 2: the results of cancellous bone grafting after EC; 1: the results of osteorchondral transplantation; 3: the results of fixation; and 1: the results of retrograde drilling. The average success rate of NT was 45%. Comparison of different surgical procedures showed that the highest average success rate is reached by excision, curettage, and drilling (ECD; 86%), followed by excision and curettage (EC; 78%) and excision alone (38%). On the basis of this systematic review, we conclude that NT and excision alone are not to be recommended in treating talar OCD. Both EC and ECD have been shown to lead to a high percentage good/excellent results. At the present time, ECD seems to be the most effective treatment strategy for osteochondral defect of the talus. Due to great diversity in the articles and variability in treatment results, however, no definitive conclusions can be drawn. Sufficiently powered randomized clinical trials with uniform methodology and validated outcome measures should be initiated to compare the outcome of surgical strategies for OCD of the talus. PMID- 12911239 TI - Arthroscopic debridement and drilling of osteochondral lesions of the talus. AB - Diagnosis of OLTs requires a high index of suspicion because these lesions are rare and the symptoms can be falsely attributed to acute or chronic ankle sprains. When no abnormality is present on plain radiographs, a bone scan or MRI can reliably identify the presence of an OLT. CT scanning can provide even better detail of the location and size of the fragment and help stage these lesions and guide treatment. Arthroscopic staging is believed to be the best method to determine treatment. In a patient without an obvious loose body, initial nonoperative treatment is warranted. When nonoperative therapy fails or when a high stage lesion is present, operative options should be explored. Arthroscopic techniques provide results that are equal to or better than management by arthrotomy and have the advantages of lower morbidity and quicker overall rehabilitation time. In most cases, arthroscopic treatment involves loose body removal and debridement and drilling of the underlying bone or drilling alone for intact lesions. Although it is unknown whether such treatment can reduce the incidence of late arthrosis in a patient who has an OLT, a recent study suggested that healing occurs and the MRI appearance of the talar dome normalizes in many patients postoperatively. PMID- 12911240 TI - The mosaicplasty technique for osteochondral lesions of the talus. AB - For years, OCD of the talus has been known as a symptomatic lesion that causes pain, recurrent synovitis, altered joint mechanism, and obstruction from loose bodies. It is a probable precursor of ankle osteoarthritis because of altered joint mechanics and recurrent synovitis. With the notable advance of diagnostic imaging and the advent of ankle arthroscopy, classification of the lesion has become standardized, which allowed for the comparison of treatment options. Arthroscopic procedures (eg, debridement, retrograde drilling, bone grafting, by nature of their minimally invasive approach, have a great advantage in treating small defects and stable OCD lesions compared with open methods. For larger osteochondral defects and unstable OCD lesions, the optimal treatment is the long term replacement and integration of type-specific hyaline cartilage. In principle, mosaicplasty autogenous osteochondral transplantation fills these criteria. The early- and medium-term results are encouraging, complete with confirmatory radiographs and histology, and hold promise for this procedure to provide lasting relief of symptoms and the prevention of ankle arthrosis. Under the current dichotomy of nonoperative and operative treatments giving satisfactory results, and few comparative studies, there is a need for a randomized, prospective study in the treatment of talar OCD to define a reproducible treatment algorithm. PMID- 12911241 TI - Osteochondral autologous transfer system. AB - The OATS technique is one of several cartilage repair procedures that exhibits promising short- to intermediate-term results in the surgical management of OLTs. Although the OATS procedure is generally reserved for salvage of failed debridement and drilling, some investigators are suggesting that it may have applications in primary surgical management of OLTs, particularly those that are associated with subchondral cysts. Long-term outcome of the OATS procedure for OLTs is not yet available. PMID- 12911242 TI - Autologous chondrocyte transplantation of the ankle. AB - Autologous chondrocyte transplantation has been used in the treatment of chondral and osteochondral lesions in a limited number of patients. The results are promising over a medium-length follow-up (2-6 years). Deeper involvement of the subchondral bone may need autologous bone grafting along with ACT. Future long term, comparative studies are needed to evaluate the efficacy and safety of the treatment. PMID- 12911243 TI - Distinguishing ankle and knee articular cartilage. AB - Degenerative changes in the tall and femoral distal cartilages of more than 2,000 tissue donors were graded based on the appearance of articular cartilage and osteophytes. In the ankle and the knee the degenerative changes increased with age; however, the rate of degeneration in the ankle was slower than in the knee. The degenerative changes in the ankle were more severe in men than in women, were predominantly bilateral, and seemed to be correlated with weight. The slower rate of change in the ankle may be caused, in part, by the biochemical and biomechanical tissue properties that distinguish ankle cartilage from that of the knee. PMID- 12911244 TI - Supramalleolar osteotomy: indications and technique. AB - The distal tibial (supramalleolar) osteotomy for the treatment of pathologic entities of the adult distal tibia and foot and ankle has received limited attention in the literature. It is technically demanding and requires an extensive and careful preoperative planning. In our experience, it has been a useful tool in the surgical armamentarium to reconstruct the normal mechanical environment in malunion preventing any long-term deleterious effects, and to shift and redistribute loads in the ankle joint to protect the articular cartilage from further degeneration. PMID- 12911245 TI - Clinical benefit of joint distraction in the treatment of ankle osteoarthritis. AB - Irrespective of underlying mechanisms, the structural changes after joint distraction and the efficacy during several years validate the concept of joint distraction in the treatment of osteoarthritis. Therefore, joint distraction in the case of severe ankle osteoarthritis at relatively young age may be a treatment of choice. In the light of increased aging, and the limited life span of an endoprosthesis, evaluation of joint distraction in the case of knee and hip osteoarthritis is justified. PMID- 12911246 TI - Arthrodesis of the ankle: arthroscopic, mini-open, and open techniques. AB - Despite recent advances in ankle arthroplasty technology, arthrodesis remains the gold standard for the operative treatment of advanced ankle arthritis. Choice of technique should be guided by the surgeon's experience and the amount of deformity present. In all cases, close attention to detail is essential in performing this procedure to ensure that the patient is provided with a well fused, stable ankle and a well-aligned plantar grade foot suitable for functional ambulation. PMID- 12911247 TI - Use of allografts in the management of ankle arthritis. AB - Reconstruction of articular cartilage defects of the tibiotalar joint remains a challenge. Although arthrodesis and total ankle arthroplasty are treatment options, we present fresh tibiotalar allografting as an alternative technique. The average age of 12 patients who underwent tibiotalar allografting was 43 years. The average follow-up was 21 months. All grafts healed at the host/donor interface. Complications included intraoperative fracture in one patient and graft collapse that required revision allografting in another. Most patients were relieved of preoperative pain and were satisfied with the procedure. Postoperative function was also significantly improved, based on questionnaire and physician assessment. Fresh tibiotalar allografting is an exciting and promising technique in the treatment of articular cartilage defects in young, active patients. PMID- 12911248 TI - Total ankle replacement. AB - Total ankle replacement is emerging as a viable treatment for symptomatic ankle arthritis that has not responded to nonoperative treatment. Most modern implant designs involve the use of a three-component, mobile-bearing implant. Although the results of the different design approaches are encouraging in limited clinical series, there is still the need for careful, long-term analyses to estimate to what extent the current designs are mimicking the biomechanics of the ankle joint. More attention must be paid to more accurate implantation techniques that result in a well-balanced ligament and allow the ligaments to act together with the replaced surfaces in a most physiologic manner. To improve the stability of the bone-implant interface over time, specific efforts should be made to better understand the extrinsic and intrinsic forces of the ankle joint, including the forces that are created by the prosthesis itself. PMID- 12911249 TI - Diagnosed diabetes and ethnic disparities in adverse health behaviors of American women. AB - Despite higher rates of some high-risk lifestyle factors in non-Hispanic black women compared to non-Hispanic white women, no data exist examining the role of diagnosed diseases. Having diabetes diagnosed might motivate women and their providers to work together to lower the women's levels of behavioral risk factors. The purpose of this study was to determine the association between diagnosed diabetes and adverse health behaviors, including smoking, alcohol consumption, and sedentary lifestyle in non-Hispanic white (n=270) and non Hispanic black (n=149) women with type 2 diabetes. Diagnosed diabetes was defined as answering "yes" to the Third US National Health and Nutrition Examination Survey question: "Have you ever been told by a doctor that you have diabetes or sugar diabetes?" Logistic regression analyses were used to determine the association of diagnosed diabetes with the adverse health behaviors. In this study, non-Hispanic black diabetic women had higher prevalences of smoking, sedentary lifestyle, and lower rates of diagnosed diabetes compared with non Hispanic white women (P<0.01). Relative to non-Hispanic diabetic white, non Hispanic diabetic black was associated respectively with 25% and 58% increased odds of smoking and sedentary lifestyle, adjusting for diagnosed diabetes and other confounding variables. Approximately 15% of alcohol consumption and 13% excess sedentary lifestyle in non-Hispanic diabetic blacks were associated with their increased rates of diagnosed diabetes relative to non-Hispanic diabetic whites. These excesses in adverse health behaviors, however, were within what can be explained by chance variation. There were non-significant trends toward less smoking and more sedentary lifestyle. Thus, diabetic women with a diagnosis generally had a worse behavioral risk profile than those without a diagnosis even after controlling multiple confounders. This shows the need for physicians to educate their diabetic patients regarding benefits of exercise and smoking avoidance. PMID- 12911250 TI - Depression in sickle cell disease. AB - PURPOSE: To assess the prevalence of depressive symptoms and examine the contribution of demographics, disease severity, and health care use variables to depressive symptoms in sickle cell patients who had been in stable health for at least one month. PATIENTS AND METHODS: Subjects were a convenience sample of 27 men and 23 women selected during a routine visit to the sickle cell clinic at Howard University Hospital. Depression was assessed using a cut-off score from the Beck Depression Inventory (BDI) and related to a variety of health outcomes. RESULTS: The results of the analyses indicate that 44% (n=22) of the sample scored within the mild to severe (>20) range of depression on the BDI. Depressed sickle cell patients were more frequently treated in emergency rooms and more likely to be hospitalized with vaso-occlusive crises. Patients more likely to be depressed were: those with low family income (<$10,000); less than high school education; female; those who had multiple blood transfusions; poor pain control; inadequate social support; hydroxyurea use; and had histories of frequent vaso occlusive crises. CONCLUSION: The prevalence of depressive symptoms in sickle cell patients is high compared to the general African American population. Our findings confirmed previous studies examining the occurrence of depression in adults with sickle cell disease. Treatment of depression should be strongly considered to improve the quality of life and probably disease course in sickle cell patients. PMID- 12911251 TI - Genetics and health disparities: fears and realities. AB - The development and testing of explanatory hypotheses about the underlying mechanisms that create health disparities among ethnic minorities will be crucial in identifying solutions for reducing the current differentials. This paper addresses the potential for using genetic information as a useful and necessary addition to approaches to measures of the "environment" in the study of the origins of health disparities. Approaches and theoretical perspectives on the integration of social science and genetic findings are discussed. PMID- 12911252 TI - Association of serum C-reactive protein and indices of body fat distribution and overweight in Mexican American children. AB - BACKGROUND: Few data have been published on the association of indices of body fat distribution and components of the insulin resistance syndrome and serum C reactive protein (CRP), an acute phase protein and putative risk factor for cardiovascular morbidity, in representative samples of total populations of children or in Hispanic Americans, who have a high prevalence of obesity and diabetes as adults. OBJECTIVE: To evaluate the association of waist-to-hip ratio (WHR) and body mass index (BMI) and components of the insulin resistance syndrome with CRP in Mexican American children and to assess the independence of the association. DESIGN: Cross-sectional survey of a large national sample, the Third National Health and Nutrition Examination Survey. PARTICIPANTS: Mexican American children aged 6-11 years. MEASUREMENTS: Body circumferences, skinfold thickness, BMI, blood pressure, and serum CRP and lipid concentrations. RESULTS: Overall, 11% of children had detectable CRP (> 0.21 mg/dL). CRP was not associated with age, gender, or birth weight. WHR showed significant positive associations with serum CRP concentration independent of BMI. BMI was also significantly associated with CRP independent of WHR. CRP was significantly associated with HDL cholesterol but not triglyceride, or systolic blood pressure, concentration after controlling for BMI. CONCLUSION: Further research is needed on the associations of serum CRP concentration with WHR and other indices of body fat distribution and obesity to elucidate the mechanisms and significance of these associations. PMID- 12911253 TI - Central serous chorioretinopathy in African Americans. AB - PURPOSE: Central Serous Chorioretinopathy (CSCR) is presumed to be less prevalent in the African American population. The purpose of this study was to compare the characteristics of CSCR in African Americans and Caucasians. METHODS: A retrospective analysis was performed. Visual acuity (VA) evaluations that were recorded included best-corrected VA at diagnosis, worst VA recorded at follow-up, and best-corrected VA at the last clinic visit. Recurrences of CSCR, frequency of laser photocoagulation, and fluorescein angiographic patterns also were evaluated. RESULTS: Of the 74 patients with CSCR, 15 (20.3%) were African American and 59 (79.7%) were Caucasian. This ethnic distribution was similar to the ethnic distribution in the entire Henry Ford Health System population. The mean VA at presentation was significantly lower in African-Americans (20/55 vs. 20/30, P=0.004) and trended towards being lower during follow-up (20/58 vs. 20/32, P=0.04) and at final examination (20/28 vs. 20/22, P=0.04). Mean length of follow-up was 21 months for both groups. CONCLUSION: The rates and spectrum of symptomatic CSCR seen at Henry Ford Health System are comparable in African Americans and Caucasians. PMID- 12911254 TI - Acculturation and psychosocial stress show differential relationships to insulin resistance (HOMA) and body fat distribution in two groups of blacks living in the US Virgin Islands. AB - The objective of this study was to determine whether acculturation and psychosocial stress exert differential effects on body fat distribution and insulin resistance among native-born African Americans and African-Caribbean immigrants living in the US Virgin Islands (USVI). Data collected from a non diabetic sample of 183 USVI-born African Americans and 296 African-Caribbean immigrants age > 20 on the island of St. Croix, USVI were studied. Information on demographic characteristics, acculturation and psychosocial stress was collected by questionnaire. Anthropometric measurements were taken, and serum glucose and insulin were measured from fasting blood samples. Insulin resistance was estimated by the homeostasis model assessment (HOMA) method. The results showed that in multivariate regression analyses, controlling for age, education, gender, BMI, waist circumference, family history of diabetes, smoking and alcohol consumption, acculturation was independently related to logarithm of HOMA (InHOMA) scores among USVI-born African Americans, but not among African Caribbean immigrants. In contrast, among USVI-born African Americans psychosocial stress was not significantly related to InHOMA, while among African-Caribbean immigrants psychosocial stress was independently related to InHOMA in models that included BMI, but not in those which included waist circumference. This study suggests that acculturation and psychosocial stress may have a differential effect on body fat distribution and insulin resistance among native-born and immigrant blacks living in the US Virgin Islands. PMID- 12911256 TI - Classification of acute respiratory disorders of all newborns in a tertiary care center. AB - OBJECTIVE: To assess the usefulness of current diagnostic criteria in the understanding of neonatal respiratory distress in a tertiary care hospital. METHODS: We prospectively studied 2824 consecutive deliveries to determine the frequency of respiratory disorders of all types. We used definitions based on standard texts, with borderline cases being classified as having the disease in question. RESULTS: Somewhat less than half of all symptomatic infants met textbook criteria for a respiratory diagnosis. Of this subset, the most common diagnosis was respiratory distress syndrome (RDS), followed by transient tachypnea of newborn (TTN), meconium aspiration syndrome (MAS), pneumonia and others. The 323 infants who fit no standard diagnosis all had self-limited conditions similar to TTN. Most (52%) were well in less than 12 hours. Those still symptomatic after 12 hours differed from the definition of TTN by having a clear chest film (38%) and/or by requiring mechanical ventilation (10%). A slight revision of the traditional diagnostic criteria allowed classification of all these cases. CONCLUSION: More than 50% of newborns with acute respiratory symptoms do not fit textbook definitions, even broad definitions which include borderline cases. The concept of TTN should be expanded to include cases with a normal chest film. In addition, we suggest adding the category "transient respiratory insufficiency of the newbom" (TRIN) for babies ventilated briefly but not demonstrably surfactant deficient or infected. This category probably includes infants with many contributing etiologies. PMID- 12911255 TI - Primary care residents' characteristics and motives for providing differential medical treatment of cervical cancer screening. AB - BACKGROUND: Cervical cancer screening rates in the United States are sub-optimal. Physician factors likely contribute to these lower rates. Previous studies provide inconclusive evidence about the association between physician characteristics and the likelihood of addressing cervical cancer. This report assesses potential mechanisms that explain why certain providers do not address cervical cancer screening. METHODS: One hundred primary care residents from various specialties were asked to indicate the preventive topics they would address with a hypothetical white female in her early 20s, who was portrayed as living a "high risk" lifestyle, and visiting her provider only for acute care reasons. RESULTS: Among the provider characteristics assessed, only residents' ethnicity was associated with the likelihood of and time spent addressing cervical cancer screening. In particular, Asian-American residents were least likely to address cervical cancer, while African-American residents were most likely. A mediation analyses revealed that perceived barriers for addressing cervical cancer accounted for this difference. CONCLUSIONS: Study results suggest that there may be cultural factors among health care providers that may account for differential referral and treatment practices. Findings from this study may help identify factors that explain why cervical cancer screening rates are not higher. PMID- 12911258 TI - The education and medical practice of Dr. James McCune Smith (1813-1865), first black American to hold a medical degree. AB - James McCune Smith (1813-1865)--first black American to obtain a medical degree, prominent abolitionist and suffragist, compassionate physician, prolific writer, and public intellectual--has been relatively neglected by historians of medicine. No biography of Smith exists to this day, though he has been the subject of several essays. Born, in his own words, "the son of a self-emancipated bond woman," and denied admission to colleges in the United States, his native land, Smith earned medical, master's, and baccalaureate degrees at Glasgow University in Scotland. On his return to New York City in 1837, Smith became the first black physician to publish articles in US medical journals. Smith was broadly involved in the anti-slavery and suffrage movements, contributing to and editing abolitionist newspapers and serving as an officer of many organizations for the improvement of social conditions in the black community. In his scientific writings Smith debunked the racial theories in Thomas Jefferson's Notes on the State of Virginia, refuted phrenology and homeopathy, and responded with a forceful statistical critique to the racially biased US Census of 1840. Frederick Douglass, Gerrit Smith, and John Brown personally collaborated with James McCune Smith in the fight for black freedom. As the learned physician-scholar of the abolition movement, Smith was instrumental in making the overthrow of slavery credible and successful. PMID- 12911259 TI - African American men, prostate cancer screening and informed decision making. AB - Prostate cancer is the second leading cause of cancer deaths in African American men. African Americans are at increased risk over other groups and have higher mortality. Since prostate cancer is highly variable among men, medical organizations are not in agreement whether men should be screened or the appropriate ages to screen. Many of these organizations recommend discussion with patients about the benefits and limitations of screening. Some of these groups support informed decision-making (IDM). Through IDM, the patient obtains all of the information about prostate cancer including risk, to make an informed choice regarding screening. Due to several factors including lowered engagement of African American men in the healthcare system, disparities in treatment, increased risk in developing and dying from the disease, as well as other cultural and structural constraints, IDM is examined and proposed as an appropriate tool for African American men. The use of IDM is discussed, along with several challenges and cautions. We conclude with recommendations and suggestions to the provider and patient to facilitate discussions regarding prostate cancer. PMID- 12911260 TI - Style watch: looking at the bigger picture. PMID- 12911261 TI - War in Iraq: medical journals and the manufacture of consent. PMID- 12911262 TI - NCI sets goal of eliminating suffering and death due to cancer by 2015. PMID- 12911263 TI - Mitochondrial-targeted drug and DNA delivery. AB - The field of mitochondrial research is currently among the fastest growing disciplines in biomedicine. Approximately 12,000 articles on mitochondria have been published since the beginning of the new millennium. What brings mitochondria into the limelight of the scientific community? Since the end of the 1980s, a series of key discoveries has been made that have rekindled the scientific interest in this long-known cell organelle. It has become increasingly evident that mitochondrial dysfunction contributes to a variety of human disorders, ranging from neurodegenerative and neuromuscular diseases, obesity, and diabetes to ischemia-reperfusion injury and cancer. Moreover, since the middle of the 1990s, mitochondria, the "power houses" of the cell, have also become accepted as the cells' "arsenal," reflecting their increasingly acknowledged key role during apoptosis. Based on these recent developments in mitochondrial research, increased pharmacological and pharmaceutical efforts have lead to the emergence of mitochondrial medicine" as a new field of biomedical research. Targeting of biologically active molecules to mitochondria in living cells will open avenues for manipulating mitochondrial functions, which may result in the selective protection, repair, or eradication of cells. This review gives a comprehensive overview of current strategies of mitochondrial targeting and their possible therapeutic applications. PMID- 12911265 TI - Inhibition of photohemolysis induced by m-chloroperbenzoic acid by metal complexes with SOD-mimetic activity. AB - Red blood cells (RBCs) are probably the most common target through the damaging action of reactive oxygen species on the cells. The photohemolysis activity of m chloroperbenzoic acid (CPBA) was concentration- and exposure time-dependent. Twenty minutes photo exposure time and 200 microm of CPBA concentration were optimum to study the effect of generated superoxide (O2-) and hydroxyl (*OH) radicals on RBCs. RBCs lysis photosensitized by CPBA was investigated in the presence of [(VL2O)(VL2H2O)]Cl6, [MnL2O]2Cl42H2O, [FeL2Cl2]Cl H2O, [CoL2Cl2]4H2O or [ZnL2Cl2]H2O respectively, where L is 2-methylaminopyridine, with SOD-mimetic activities with the aim of ascertaining their protective activity towards the photo induced cell damage. The decrease of photolytic activity caused by these complexes was concentration-dependent and the maximum percentage of protective activity was 75, 70, 68, 57 or 24% for [(VL2O)(VL2H2O)]Cl6, [MnL2O]2Cl4 2H2O, [FeL2Cl2]Cl H2O, [CoL2Cl2]4H2O or [ZnL2Cl2]H2O complex respectively, against the cell irradiated without addition of metal complexes. The comparison between the decrease of photolytic activity caused by these complexes and their SOD-mimetic activity of these metal complexes showed an appreciable correlation. PMID- 12911264 TI - Ethosomes: new prospects in transdermal delivery. AB - Ethosomes are noninvasive delivery carriers that enable drugs to reach the deep skin layers and/or the systemic circulation. Although ethosomal systems are conceptually sophisticated, they are characterized by simplicity in their preparation, safety, and efficacy--a combination that can highly expand their application. Ethosomes are soft, malleable vesicles tailored for enhanced delivery of active agents. This article reviews work carried out in vitro, in vivo, in animal models, and in humans with various ethosomal systems incorporating a wide range of drugs. Because of their unique structure, ethosomes are able to encapsulate and deliver through the skin highly lipophilic molecules such as cannabinoids, testosterone, and minoxidil, as well as cationic drugs such as propranolol and trihexyphenidil. Results obtained in a double-blind two-armed randomized clinical study showed that treatment with the ethosomal acyclovir formulation significantly improved all the evaluated parameters. Preliminary studies with plasmids and insulin revealed that the ethosomal carrier may be used for enhanced delivery of these agents. In further work, the ethosomal technology was broadened to introduce agents into cultured cells and microorganisms. Enhanced delivery of bioactive molecules through the skin and cellular membranes by means of an ethosomal carrier opens numerous challenges and opportunities for the research and future development of novel improved therapies. PMID- 12911266 TI - Characterization of the high resolution ESR spectra of the methoxyl radical adducts of 5-(diethoxyphosphoryl)-5-methyl-1-pyrroline N-oxide (DEPMPO). AB - Spin-trapping investigators are largely limited by the instability of the radical adducts. Spin trap 5-(diethoxyphosphoryl)-5-methyl-1-pyrroline N-oxide (DEPMPO) forms very stable alkoxyl radical adducts. However, the presence of two chiral centers in the DEPMPO alkoxyl radical adduct results in two diastereomers with distinctive ESR spectra, which complicates the interpretation of the ESR spectra. We have analyzed the high resolution ESR spectra of the DEPMPO/*OCH3 radical adduct. DEPMPO/*OCH3 has been synthesized by the nucleophilic addition of alcohols to DEPMPO. The electron spin resonance (ESR) spectrum of DEPMPO/*OCH3 in oxygen-free methanol solution reveals superhyperfine structure with hyperfine coupling constants as small as 0.3G. In order to simplify the analysis of the electron spin resonance (ESR) spectrum, we synthesized the DEPMPO/*OCD3 radical adduct. Computer simulation of the DEPMPO/*OCD3 ESR spectrum revealed two diastereomers. Hyperfine coupling constants of gamma-protons and 17O from the OCH3 group were also determined. ESR spectra of DEPMPO/*OCH3 in phosphate buffer have also been characterized. The presence of specific hyperfine couplings from the -OCH3 group can be used for the unambiguous identification of the DEPMPO/*OCH3 radical adducts. We suggest that the analysis of high resolution ESR spectra can be used for the unambiguous characterization of DEPMPO radical adducts. PMID- 12911267 TI - Effects of endurance training on three superoxide dismutase isoenzymes in human plasma. AB - The effects of endurance training and acute exhaustive exercise on plasma levels of three superoxidedismutase (SOD) isoenzymes and the ability of superoxide generation in neutrophils were studied. Eighteen healthy male students, aged 17 22 years, who volunteered for this study, underwent three months of endurance training in swimming or running. Before and after the training course, they performed acute exercise and blood samples were collected before and after this exercise. The endurance training significantly increased maximal oxygen uptake (VO2max) in all subjects. Neither the endurance training nor the acute exercise affected the plasma CuZn-SOD level. Acute exercise after the training, but not before the training, increased both the plasma Mn-SOD and extracellular SOD (EC SOD) levels by 33.6 and 33.5%, respectively. The training decreased the EC-SOD level at rest by 22.2%. Acute exercise after the training, but not before the training, increased the plasma lipid peroxide level, suggesting higher oxidative stress in trained subjects during exhaustive exercise. The ability of neutrophils to generate superoxide was increased by the acute exercise, but induction of the superoxide was suppressed after training. These results indicate that EC-SOD levels were changed in a different manner from the CuZn-SOD and Mn-SOD: it was decreased by training but was increased by acute exercise, suggesting that endurance training increases the reserve of EC-SOD in tissues. The results also suggest the possibility of plasma EC-SOD assay as a new index of endurance training. PMID- 12911268 TI - Cardioprotective and antioxidant effects of apomorphine. AB - Apomorphine is a potent antioxidant that infiltrates through biological membranes. We studied the effect of apomorphine (2 microM) on myocardial ischemic reperfusion injury in the isolated rat heart. Since iron and copper ions (mediators in formation of oxygen-derived free radicals) are released during myocardial reperfusion, apomorphine interaction with iron and copper and its ability to prevent copper-induced ascorbate oxidation were studied. Apomorphine perfused before ischemia or at the commencement of reperfusion demonstrated enhanced restoration of hemodynamic function (i.e. recovery of the work index (LVDP x HR) was 69.2 +/- 4.0% with apomorphine pre-ischemic regimen vs. 43.4 +/- 9.01% in control hearts, p < 0.01, and 76.3 +/- 8.0% with apomorphine reperfusion regimen vs. 30.4 +/- 11.1% in controls, p < 0.001). This was accompanied by decreased release of proteins in the effluent and improved coronary flow recovery in hearts treated with apomorphine after the ischemia. Apomorphine forms stable complexes with copper and with iron, and inhibits the copper-induced ascorbate oxidation. It is suggested that these iron and copper chelating properties and the redox-inactive chelates formed by transition metals and apomorphine play an essential role in post-ischemic cardioprotection. PMID- 12911269 TI - Comparison of antioxidant activity between aromatic indolinonic nitroxides and natural and synthetic antioxidants. AB - In view of the possible employment of nitroxide compounds in various fields, it is important to know how they compare with other synthetic antioxidant compounds currently used in several industries and with naturally occurring antioxidants. To address this issue, the antioxidant activity of two aromatic indolinonic nitroxides synthesized by us was compared with both commercial phenolic antioxidants (BHT and BHA) and with natural phenolic antioxidants (alpha hydroxytyrosol, tyrosol, caffeic acid, alpha-tocopherol). DPPH radical scavenging ability and the inhibition of both lipid and protein oxidation induced by the peroxyl-radical generator, AAPH, were evaluated. The results obtained show that overall: (i) the reduced forms of the nitroxide compounds are better scavengers of DPPH radical than butylated hydroxyanisole (BHA) and butylated hydroxytoluene (BLT) but less efficient than the natural compounds; (ii) the nitroxides inhibit both linolenic acid micelles and bovine serum albumin (BSA) oxidation to similar extents as most of the other compounds in a concentration-dependent fashion. Since the aromatic nitroxides tested in this study are less toxic than BHT, these compounds may be regarded as potential, alternative sources for several applications. The mechanisms underlying the antioxidant activity of nitroxides were further confirmed by UV-Vis absorption spectroscopy experiments and macroscale reactions in the presence of radicals generated by thermolabile azo compounds. Distribution coefficients in octanol/buffer of the nitroxides and the other compounds were also determined as a measure of lipophilicity. PMID- 12911270 TI - Sex-dependent antioxidant enzyme activities and lipid peroxidation in ageing mouse brain. AB - We investigated whether oxidant status and antioxidant enzyme activities during ageing of mouse brain are regulated in sex-dependent manner. In the homogenate from the brain of 1, 4, 10 and 18 months old male and female CBA mice, lipid peroxidation (LPO), total superoxide dismutase (tSOD), catalase (CAT) and glutathione peroxidase (Gpx) were determined. LPO was age- and sex-related, favoring males over females throughout the lifespan with the peak in both sexes at 10 months of age. Throughout ageing, no difference in tSOD activity between male and female brains was observed, except in immature 1 month old mice. Gender related difference in Gpx activity was observed, with higher level in females comparing to males, reaching statistical significance in senescent (18 months old) animals. CAT activity was drastically changed with ageing in both the male and female brain. We found different age associated trends in CAT activity in males and females: decreased with age in males and increased with age in females. Taken together, the present findings indicate that brains of female mice have lower oxidant and higher antioxidant capacity mostly related to CAT and to a lesser extent to Gpx activity. PMID- 12911271 TI - Effects of iNOS-related NO on hearts exposed to liposoluble iron. AB - Inducible nitric oxide synthase (iNOS) protects heart against ischemia/reperfusion injury. However, it is unknown whether the beneficial effects of iNOS are mediated by the interaction of NO with radical oxygen species (ROS). To address this issue, we examined the effects of liposoluble iron-induced ROS generation in isolated perfused hearts from rats treated with lipopolysaccharide (LPS). LPS administration (10 mg/kg, i.p., 6 h before heart removal) induced iNOS expression and increased NO production as indicated by a 3 fold elevation of nitrite level in coronary effluents relative to control hearts. An enhanced expression of hemeoxygenase 1 protein was also observed in septic hearts compared to control. Iron-induced perfusion and contractile deficits were ameliorated by LPS with more important coronary than myocardial benefits. In iron loaded hearts, oxidative stress as measured by the 2,3 dihydroxybenzoic acid/salicylic acid concentration ratio in cardiac tissue was 23% lower in septic than in control heart although the difference did not reach significance. In addition, the presence of the NO synthase inhibitor N-nitro-L-arginine in the perfusion medium totally blocked NO production but did not reverse the protective effects of LPS. The results indicate that LPS protects from iron-induced cardiac dysfunction by mechanisms independent on ex vivo NO production and suggest that NO acts as a trigger rather than a direct mediator of the cardioprotective effects of LPS in heart exposed to iron. PMID- 12911272 TI - Non-invasive analysis of reactive oxygen species generated in NH4OH-induced gastric lesions of rats using a 300 MHz in vivo ESR technique. AB - Free radicals are reportedly involved in mucosal injury, including NH4OH-induced gastric lesions, but the kind, location and origin of radical generation have yet to be clarified. We developed the non-invasive measurement of reactive oxygen species (ROS) in stomach, and applied to mucosal injury. NH4OH-induced gastric lesions were prepared in rats, which were then given a nitroxyl probe intragastrically or intravenously, and the spectra of the gastric region were obtained by in vivo 300 MHz electron spin resonance (ESR) spectroscopy. The spectral change of the nitroxyl probe administered intragastrically was significantly enhanced 30 min after NH4OH administration, but no change occurred when the probe was given by intravenous injection. The enhanced change was confirmed to be due to *OH generation, because it was completely suppressed by mannitol, catalase and desferrioxamine (DFO), and was not observed in neutropenic rats. NH4OH-induced neutrophil infiltration of the gastric mucosa was suppressed by intravenous injection of superoxide dismutase (SOD) or catalase, or by administration of allopurinol. The present study provided the direct evidence in NH4OH-treated living rats that *OH produced from O2*- derived from neutrophils caused gastric lesion formation, while O2*- or H2O2 derived from the xanthine oxidase system in endothelial cells was involved in neutrophil infiltration. PMID- 12911273 TI - Confirmation of superoxide generation via xanthine oxidase in streptozotocin induced diabetic mice. AB - Reactive oxygen species (ROS) may play key roles in vascular inflammation and atherogenesis in patients with diabetes. In this study, xanthine oxidase (XO) system was examined as a potential source of superoxide in mice with streptozotocin (STZ)-induced experimental diabetes. Plasma XO activity increased 3-fold in diabetic mice (50 +/- 33 microU/ml) 2 weeks after the onset of diabetes, as compared with non-diabetic control mice (15 +/- 6 microU/ml). In vivo superoxide generation in diabetic mice was evaluated by an in vivo electron spin resonance (ESR)/spin probe method. Superoxide generation was significantly enhanced in diabetic mice, and the enhancement was restored by the administration of superoxide dismutase (SOD) and 4,5-dihydroxy-1,3-benzene disulfonic acid (Tiron), which was reported to scavenge superoxide. Pretreatment of diabetic mice with XO inhibitors, allopurinol and its active metabolite oxipurinol, normalized the increased superoxide generation. In addition, there was a correlation (r = 0.78) between the level of plasma XO activity and the relative degree of superoxide generation in diabetic and non-diabetic mice. Hence, the results of this study strongly suggest that superoxide should be generated through the increased XO seen in the diabetic model mice, which may be involved in the pathogenesis of diabetic vascular complications. PMID- 12911274 TI - Induction of the differentiation of HL-60 promyelocytic leukemia cells by L ascorbic acid. AB - The present study was undertaken to examine the effect of L-ascorbic acid (LAA) on the growth of HL-60 promyelocytic leukemia cells, besides induction of apoptosis. LAA (> or = 10(-4) M) was found to markedly inhibit the proliferation of HL-60 in liquid culture and clonogenicity in semisolid culture. Moreover, LAA treated HL-60 showed activity to produce chemiluminescence and expressed CD 66b cell surface antigens, indicating that LAA induces the differentiation of HL-60 mainly into granulocytes. The results are supported by morphological changes of LAA-treated HL-60 into segmented neutrophils. Therefore, the inhibitory effect of LAA on the growth of HL-60 cells seems to arise from the induction of differentiation. To assess the potential role of LAA, cells were exposed to oxygen radical scavengers in the absence or presence of LAA. Catalase abolished and superoxide dismutase promoted LAA-induced differentiation of HL-60. Thus, H2O2 produced as a result of LAA treatment seems to play a major role in induction of HL-60 differentiation. PMID- 12911275 TI - Effect of vitamin E on serum aminotransferase and thioredoxin levels in patients with viral hepatitis C. AB - OBJECTIVES: Oxidative stress induces cellular responses such as cell death, gene activation and cell proliferation, in the liver. Vitamin E (Vit. E) has been found to protect the liver against oxidative stress in animal experiments. Thioredoxin (TRX) is a stress inducible, multifunctional protein, secreted during oxidative stress. This study evaluated effects of Vit. E on serum TRX and aminotransferase levels in hepatitis C virus (HCV) patients, partly non responsive to initial interferon (IFN), with higher than average level of serum alanine aminotransferase (ALT) after receiving anti-inflammatory drug treatment. METHODS: Seventeen HCV patients (male = 3; female = 14) of age 62 +/- 7.65 years receiving anti-inflammatory drug therapy, at least 6 months prior to Vit. E administration, were given d-alpha-tocopherol 500 mg/day, orally, for a period of 3 months. ALT, aspartate aminotransferase (AST), TRX and Vit. E were measured at 0, 1, 2 and 3 months and 1 month after end of treatment. As controls, the same patients biochemical data, 3 months from the start of therapy were used. Patients were divided into three categories: total patients "T", low ALT group "L" (ALT < 70 IU/l) and high ALT group "H" (ALT > 70 IU/l), respectively. RESULTS: The ALT level was lowered, significantly in group H, in the 1st, 2nd, 3rd and 1-month post therapy, compared to the initial value. But group L showed little or no change in ALT. Post Vit. E therapy, in groups T and H, the TRX level was elevated but remained below initial levels, whereas in group L, TRX level remained significantly lower than the pretreatment value. Groups T and L, showed significant reduction (p < 0.05) in serum TRX levels in the 2nd and 3rd month. Group H showed a tendency towards TRX reduction, but not significantly. Serum Vit. E levels increased significantly (p < 0.0001) from the 1st to 3rd month in all three T, H and L groups. CONCLUSION: Oxidative stress induced liver damage is reduced by Vit. E in patients with viral hepatitis C, particularly those with initial ALT levels > 70 IU/l. Vit. E treatment causes reduction of oxidative stress markers as TRX and ALT in sera. Therefore, Vit. E can act as a supportive therapy to combat liver damage caused by oxidative stress, in such patients with continuously high levels of ALT even after anti-viral and anti-inflammatory drug therapy. PMID- 12911276 TI - Simultaneous detection of the antioxidant and pro-oxidant activity of dietary polyphenolics in a peroxidase system. AB - The ability to reduce the peroxidase (myeloglobin/H2O2)-generated ABTS*+ [2,2' azinobis-(3-ethylbenzthiazoline-6-sulfonic acid) radical cation] has been used to rank the antioxidant activity of various agents including dietary flavonoids and chalcones. Surprisingly, we found that in the presence of catalytic concentrations of the phenol B-ring containing flavonoids, apigenin, naringenin and the chalcone phloretin, the formation of the ABTS*+ was initially increased. The enhanced formation of the ABTS*+ was attributed to the peroxidase/H2O2 mediated generation of polyphenolic phenoxyl radicals that were able to co oxidize ABTS. The relative ABTS*+ generating ability of these dietary polyphenolics correlated with their ability to co-oxidize NADH to the NAD* radical with the resultant generation of superoxide. This pro-oxidant activity was not observed for either luteolin or eriodyctiol, which are B-ring catecholic analogues of apigenin and naringenin, respectively, suggesting that these antioxidants are incapable of the transition metal-independent generation of reactive oxygen species. This pro-oxidant activity of the polyphenolics therefore needs to be taken into account when quantifying antioxidant activity. PMID- 12911277 TI - A causal association between Haemophilus influenzae type b (Hib) vaccine and diabetes. PMID- 12911278 TI - Significance of anti-oxidized LDL antibody and monocyte-derived microparticles in anti-phospholipid antibody syndrome. AB - Monocytes, platelets, endothelial cells and oxidized LDL could be very important in development of vascular complication in thrombotic diseases. We measured and compared the levels of plasma monocyte-derived microparticles (MDMPs), platelet derived microparticles (PDMPs), and anti-oxidized LDL antibody, to develop a better understanding of their potential contribution to vascular complications in antiphospholipid antibody syndrome (APS). The concentration of MDMP in APS patients was significantly higher (p < 0.01) than that in normal subjects and SLE patients. When levels of PDMPs and plt-P-selectin were compared between the control and APS patients, levels of PDMPs and plt-P-selectin were significantly higher (p < 0.01 for each) in APS patients than in controls. In addition, these levels of platelet activation markers correlated with MDMP in APS patients. Twenty one of the 37 APS patients (56.8%) had elevated levels of anti-oxLDL antibody. In addition, a significant increase in MDMP was observed in anti-oxLDL antibody-positive APS patients (p < 0.01). These findings suggest that elevated MDMPs may be a sign of vascular complication in APS patients, particularly those who are detected anti-oxLDL antibodies. PMID- 12911279 TI - Evidence for cytokine dysregulation in multiple sclerosis: peripheral blood mononuclear cell production of pro-inflammatory and anti-inflammatory cytokines during relapse and remission. AB - We investigated circulating anti-inflammatory and pro-inflammatory cytokines, and their ex vivo PBMC production in the absence or presence of the neuroantigens myelin basic protein (MBP) and myelin oligodendrocyte glycoprotein (MOG) and T cell mitogen (PHA) in MS patients in relapse and remission, patients with other neurological disorders (OND) and normal healthy controls. MS patients in relapse exhibited significantly increased PBMC production of TNF-alpha spontaneously compared with MS remission and healthy controls and with MBP compared with MS remission. Patients in relapse had significantly increased spontaneous, PHA- and MBP-induced PBMC IL-1beta production compared with remission MS, and was increased compared (PHA only) with OND and healthy controls. In relapse there was also significantly increased PBMC IFN-gamma production (PHA only) compared with remission and a significantly lower production of biologically active TGF-beta1 (PHA only) compared with remission MS and OND. In contrast, MS patients in remission produced significantly less spontaneous and MBP-induced TNF-alpha, spontaneous, PHA- and MBP-induced IL-1beta and PHA-induced IFN-gamma together with increased production of biologically active TGF-beta1. MOG non-specifically increased PBMC TNF-alpha and IL-1beta production in all groups. Pro-inflammatory cytokines in corresponding plasma samples were undetectable whilst the concentration of biologically active TGF-beta1 was the reverse of ex vivo PBMC findings. The increase in biologically active TGF-beta1 production ex vivo in OND patients, despite active disease, compared with the low level in the MS relapse may indicate a regulatory defect in MS. We conclude that the balance between biologically active TGF-beta1 and the pro-inflammatory TNF-alpha, IL-1beta and IFN-gamma is dysregulated during MS relapse-remission and that normal counter regulatory mechanisms during the relapse phase are defective. PMID- 12911280 TI - Endocrine pancreas histology of congenic BB-rat strains with reduced diabetes incidence after genetic manipulation on chromosomes 4, 6 and X. AB - Congenic BB.SHR rat strains were established by crossing of spontaneously diabetic BB/OK rats and diabetes-resistant SHR rats. Chromosomal regions on which the genes Iddm 4 (BB.6s), Iddm6 (BB.Xs) and Iddm 2 (BB.LL) are located were exchanged. As a result of genetic manipulation diabetes incidence was markedly reduced from 80% in BB/OK to 50% in BB.SHR (Chr. X), to 14% in BB.SHR (Chr. 6) and to 0% in BB.LL rats. Pancreata of these newly generated BB.SHR rats were investigated histologically. In newly diagnosed diabetic rats of congenic strains pancreatic insulin content (BB.6s: p < 0.05; BB.Xs p < 0.01) and relative volume of insulin-positive cells (BB.Xs: p < 0.001) were significantly higher than in BB/OK rats. The degree of insulitis was not different in 90-day-old and newly diagnosed diabetic animals. Surprisingly, in 30-day-old rats we observed an increase of the degree of insulitis with decreasing diabetes incidence. We suppose that by an earlier occurrence of the immunological beta-cell destruction, a part of the animals is able to develop a secondary diabetes resistance. The exchange of the BB-lymphopenia gene by that of SHR-rats prevented the development of hyperglycaemia without altering the auto-reactive immune response, which could be observed in all animals investigated. PMID- 12911281 TI - Frequency of anti-AChR epsilon subunit-specific antibodies in MG. AB - A definite diagnosis of myasthenia gravis (MG) relies heavily on acetylcholine receptor (AChR) antibody testing. The relatively high number of antibody-negative patients therefore, causes frequent uncertainty in confirming the diagnosis. We evaluated the sensitivity and specificity of a new, commercially available AChR antibody test that uses an approximately equal mixture of AChR from TE671-epsilon (adult type) and TE671-gamma (fetal type) cells. This assay was used to re examine 365 seronegative MG sera in which AChR antibody had not been detected by the standard assay that uses fetal type AChR. The new assay detected anti-AChR antibodies in 17 (15.5%) of 110 patients with ocular type and in 33 (12.9%) of 255 patients with generalized type MG. Anti-AChR epsilon subunit-specific antibodies were present in 13.7% of the patients in whom no AChR antibody had been detected by the standard assay, showing an increase from 79 to 82% in overall diagnostic sensitivity. PMID- 12911282 TI - Autoimmunity as the most frequent cause of idiopathic secondary adrenal insufficiency: report of 111 cases. AB - The origin of the isolated secondary adrenal insufficiency is unknown in most cases. An observation of a group of over 100 patients with secondary adrenal insufficiency and coexisting autoimmune abnormalities suggests that autoimmunity could be a frequent cause of idiopathic secondary failure, similarly as in Addison's disease. We studied 176 patients with idiopathic isolated secondary adrenal insufficiency. The methods included clinical examination and measurements of pituitary, adrenal, thyroid and gonadal hormones in all the cases. Since thyroid autoimmunity has been the most frequent finding in Addison's disease we have also chosen thyroid autoantibodies as markers of an autoimmune process in our material. Anti-peroxidase, anti-microsomal and anti-thyroglobulin autoantibodies were determined in 151 patients. In 111 out of 151 patients (73%) coexisting autoimmune diseases and/or presence of thyroid autoantibodies were detected. The most frequent autoimmune diseases associated with secondary adrenal insufficiency were primary hypothyroidism, hyperthyroidism (mainly in the past) and premature ovarian failure. Thyroid autoantibodies, especially antiperoxidase autoantibodies, were present in 106 patients. Thus, coexistence of isolated secondary adrenal insufficiency with some autoimmune disorders in 73% of the patients under study suggests that autoimmunity is the most frequent cause of the idiopathic form of this disease. PMID- 12911284 TI - Strain dependent rat iNOS promoter activity--correlation to identified WT1 transcription factor binding site. AB - The free radical nitric oxide (NO) has been implicated in cytokine mediated destruction of rat beta-cells in islets of Langerhans. Cytokine mediated NO production is associated with increased expression of the inducible nitric oxide synthase (iNOS). We have previously shown a strain dependent difference between Wistar Kyoto (WKY) and Brown Norway (BN) rats of IL-1beta mediated destruction of islets of Langerhans to be related to expression levels of iNOS and NO production. The aim of the present study was to clone and screen the iNOS gene promoter region from WKY and BN rats for polymorphisms and to functionally test such nucleotide differences. Within the total 2077 bp sequenced from both rat strains we identified three polymorphisms in two separate areas: (i) a GT-repeat polymorphism linked to (ii) a C/T polymorphisms, leading to a WT1 binding site approximately 1650bp upstream the BN iNOS promoter and (iii) a G/A SNP in exon 1. Apart from these polymorphisms the homology between all published rat iNOS sequences including the presently described are about 96%. Promoter activity was detected for both genes in a luciferase assay followed cloning of 2012 bp fragments and transient transfection into RIN cells. For both strains IL-1beta induced dose-dependent activity and strain dependent iNOS promoter activity was demonstrated when WT1 was co-expressed. To our knowledge, this is the first demonstration of functional WT1/iNOS promoter interaction. We conclude that the iNOS promoter is strain-dependently regulated which may relate to quantitatively as well as qualitatively strain dependent differences in transcription factor expression, in this study exemplified by WT1. PMID- 12911283 TI - Association of transporter associated with antigen processing genes with Behcet's disease in Japanese. AB - Contribution of transporter associated with antigen processing (TAP) genes to the pathogenesis of Behcet's disease (BD) was studied. Restriction fragment length polymorphic analysis of TAP genes was carried out in 46 Japanese patients with BD and 95 healthy subjects. There were no significant differences in allele frequencies of TAP1 and TAP2 genes between whole patients with BD and control population. No significant differences in the frequencies of TAP alleles were observed, when patients of BD with complete type or incomplete type were compared with control population, respectively. An allele frequency of TAP2C was, however, slightly but significantly high in patients with BD who had symptom of erythema nodosum (24.1%) as compared to the control group (11.6%). [p < .05, RR = 2.4]. The allele frequency of TAP2C was slightly high in HLA*B5101 positive patients with BD (28.6%) as compared to HLA*B5101 negative patients (10.9%), but the difference did not reach statistical significance. The absence of genotype TAP2B/C was observed in whole patients group, though it was present in control subjects (14.7%). [p = 0.003, RR = 0.06]. A genotype frequency of TAP2C/H was high in patients with BD who had symptom of skin lesions (7.5%) as compared to the control group (0.0%). [p = 0.03, RR = 15.4]. These results suggest the possibility that TAP molecule play some part in formation of skin lesion, such as erythema nodosum in BD in Japanese. PMID- 12911285 TI - Polymorphisms of HLA class II predispose children and adolescents with type 1 diabetes mellitus to autoimmune thyroid disease. AB - To determine the prevalence of autoimmune thyroid disease (AITD) in children and adolescents with type 1 diabetes mellitus (DM), and assess whether the development of AITD is correlated with specific DQ-A and DQ-B loci of the HLA class II antigens, we analyzed thyroid function using anti-thyroid antibodies and HLA-DQ-A and -DQ-B polymorphisms in 69 patients with type 1 DM, in 75 normal healthy controls, and in 21 patients with AITD but without type 1 DM. Eighteen patients (26%) in the diabetic patients had AITD. In the diabetic patients, DQA1*0301 and DQB1*0302 occurred more frequently than in controls [DQA1*0301: OR = 1.939, 95% CI = 1.210-3.109 (P = 0.008, P(c) (corrected P) < 0.05); DQB1*0302: OR = 2.558, 95% CI = 1.354-4.832 (P = 0.005, P(c), > 0.05)]. Compared with controls, non-diabetic subjects with AITD showed higher frequency of DQA1*0301 (P(c), < 0.05) and DQB1*0601 (P(c) > 0.05), but these alleles were not contributing factors in the development of AITD in diabetic patients. In diabetic patients, DQB1*0201, known as susceptible allele of type 1 DM was not a contributing factor in the development of AITD in diabetic patients. Unlike DQB1*0201, DQB1*0401 was more frequently found in diabetic patients with AITD than in controls [OR = 4.053, 95% CI = 1.607-10.221 (P = 0.0017, P(c) < 0.05)] or than in non-diabetic AITD patients [OR = 15.769, 95% CI = 1.905-130.530(P = 0.002, P(c) < 0.05)]. In non-diabetic subjects, DQB1*0401 did not provide susceptibility for AITD. Our results suggest that HLA DQB1*0401 is a predisposing genetic marker for the development of AITD in patients with type 1 DM in Korea. PMID- 12911286 TI - Disruption of the cytoskeleton after apoptosis induction with autoantibodies. AB - F-actin cleavage was studied in PBMC after treatment with anti-dsDNA antibodies. Significant changes in F-actin disruption detected by decrease of FITC-phalloidin staining occurred after apoptosis induction with anti-dsDNA antibodies (p < 0.006). Despite of similar F-actin disruption, the switch of phosphatidylserine (PS) to the outer leaflet of the cell membrane as detected by annexin V binding was lower after anti-dsDNA antibody than without antibody treatment (58.4 +/- 11.0% vs. 81.9 +/- 7.7%). F-actin disruption was accompanied by activation of caspase 3 within the cytoplasm (r = -0.92599; p < 8.87446 x 10-(10)) under both conditions with and without autoantibodies. These findings indicate that anti dsDNA antibody-induced apoptosis is more marked within the cell than upon the cell surface. The diminished externalization of PS might result in a decreased phagocytosis. Thereby, the reduced clearance of apoptotic cells could induce autoantibody production possibly against epitopes which arise due to the apoptotic disruption of cells. PMID- 12911287 TI - Surgical viewpoints for the definitive treatment of lung cancer. AB - Surgery remains a central pillar in the treatment of lung cancer. To optimize surgical interventions, careful preoperative assessment is necessary. Pulmonary status and cardiac status are the main risks to be considered. After operability has been established, resectability is assessed by staging the lung cancer. Surgery offers a variety of tools to accomplish complete staging before resection. Successful resection is defined as the complete removal of the cancer. To accomplish this goal, a multidisciplinary approach is evolving rapidly. For patients with nonoperable cancer, surgical techniques have been developed to manage airway obstructions and to drain effusions. PMID- 12911288 TI - The role of radiotherapy and chemotherapy for curative management of medically inoperable and stage III nonsmall cell lung cancer, and radiotherapy for palliation of symptomatic disease. AB - Radiotherapy has an expanding role in all phases of treatment of nonsmall cell lung cancer. Evolutions in technique, such as three-dimensional conformal radiotherapy, hold the promise for more effective treatment of patients with early stage disease who are not candidates for surgical intervention. Multimodality therapy for patients with locally advanced disease is evolving rapidly, with evidence accruing as to the optimal schedules and doses of radiotherapy and combination chemotherapy. Palliative dose schedules are being refined that maximize patient comfort while providing substantial symptom relief. PMID- 12911289 TI - Combined modality therapy of early stage nonsmall cell lung cancer. AB - Therapy for locally advanced NSCLC has evolved into a multidisciplinary effort. Patients who are considered for this approach should undergo rigorous testing to accurately stage their disease. Patients with pleural effusions (with rare exception) are not candidates for intensive combined modality therapy. Appropriate patients for combined modality therapy should have a good performance status (generally Zubrod 0 or 1), adequate pulmonary function, absence of significant heart, lung, or other medical diseases, and be appropriate candidates for combination chemotherapy and thoracic surgery or thoracic radiotherapy. Several lessons can be learned from looking broadly at the phase II and phase III combined modality experience. The available data do not support the routine use of postoperative therapy in patients with completely resected disease. Treatment with chemotherapy before surgery or radiation has demonstrated survival benefit in patients with stage III disease. The French phase III trial of induction chemotherapy in patients with early stage disease found an 11-month improvement in overall survival (P = 0.15) and a significant increase in the risk of death for patients with stage I and II disease. The ongoing U.S. intergroup trial (SWOG 9900) and European trials will help to further define the role of chemotherapy in patients with clinical stage IB, II and IIIA NSCLC. Clinical trials should be conducted to compare preoperative chemoradiotherapy with preoperative chemotherapy. The recently completed intergroup 0139 trial (chemoradiation followed by surgery or not) should help to define whether surgery and radiation are required in the management of stage IIIA NSCLC. Finally, further improvement in survival with the use of "newer" cytotoxic agents seems unlikely as phase III trials in metastatic NSCLC have not demonstrated marked superiority over cispiatin-based regimens. Ongoing trials are assessing the incorporation of newer, biologic-based "targeted" therapies. Despite the dismal findings of trials of postoperative therapy, many patients continue to have surgery as their initial treatment followed by postoperative therapy. In contrast, trials with induction treatment seem to offer improved survival. It is time for a true multidisciplinary approach to the treatment of locally advanced NSCLC. Pulmonary physicians, thoracic surgeons, medical oncologists, and radiation oncologists should meet before the initiation of treatment to plan the most appropriate therapy for the individual patient. PMID- 12911290 TI - The role of chemotherapy in the treatment of unresectable stage III and IV nonsmall cell lung cancer. AB - Modern platinum-based combination chemotherapy has played a major role in the therapeutic approach to unresectable stage III and IV NSCLC. Randomized phase III trials clearly documented a survival as well as palliative benefit to treatment in patients with stage IV NSCLC who have a good PS (PS 0-1). The optimal therapeutic approach in patients with poor PS (PS 2) has not yet been defined. Recent trials that focused on the elderly suggested that they receive benefits from chemotherapy that are similar to their younger counterparts. The benefit from chemotherapy seems to occur early (initial 3 to 4 cycles) and prolonged therapy is not indicated. Second-line therapy that is administered upon progression was shown to provide survival and palliative benefits. In unresectable stage III NSCLC, the addition of chemotherapy to TRT improves long term survival and has the potential to cure a minority of patients. Although sequential and concurrent chemoradiotherapy approaches have improved survival in phase III trials, concurrent strategies seem superior in comparative trials. New techniques in radiation therapy, such as three-dimensional treatment planning, may allow safer administration of both modalities concurrently and allow higher doses of TRT to be delivered. In unresectable stage III and stage IV NSCLC, the role of the new "targeted" therapies is currently being defined in several randomized, phase III trials. It is imperative that physicians who care for patients with advanced NSCLC be aware of these trials and attempt to enroll their patients, if possible. It is only through the successful and timely completion of well-designed clinical trials that we will advance our knowledge of improved treatment options for our patients with this disease. PMID- 12911291 TI - Palliation of endobronchial disease: flexible and rigid bronchoscopic options. AB - Many patients with lung cancer develop airway obstruction and hemoptysis. Endoscopic palliation can relieve symptoms and improve the quality of life for many patients. Patient factors, lesion characteristics and location, and regional expertise are important to consider when planning therapy. Cost, convenience, time to relief of symptoms, duration of palliation, and the need for serial procedures should also be considered. Further advances in technology will likely allow better palliation in the future. A wide armamentarium of modalities and careful communication with the patient's other providers are essential for optimal patient outcome. PMID- 12911292 TI - Altering risk in patients who smoke. AB - Because of the inherent difficulty of quitting smoking and the enormous health and societal burden of smoking, a multitude of interventions have been developed and tested for their efficacy in sustaining abstinence in smokers. Although most smokers' attempts to quit on their own end in failure, with 12-month abstinence rates as low as 5.7%, several psychosocial and pharmacological interventions have been noted for substantially increasing, even doubling or tripling abstinence rates. Given the substantial costs of treating illnesses caused by smoking, even a 1% increase in abstinence rates is notable for its public health benefit. Moreover, research has indicated that it may take the majority of smokers several attempts at quitting before total continuous abstinence is achieved. Thus any quit attempt should be construed as a step forward in the direction of sustained abstinence. The relative ease with which clinician-initiated effective treatments can be implemented. and evidence that many smokers would like to stop smoking and look to health-care practitioners for guidance and motivation, suggest that pulmonary medicine specialists can have a substantial impact on morbidity and mortality associated with smoking. PMID- 12911293 TI - Native state proline isomerization: an intrinsic molecular switch. PMID- 12911294 TI - Identification of the catalytic nucleophile of the family 29 alpha-L-fucosidase from Sulfolobus solfataricus via chemical rescue of an inactive mutant. AB - We have recently reported that a functional alpha-L-fucosidase could be expressed by a single insertional mutation in the region of overlap between the ORFs SSO11867 and SSO3060 of the hyperthermophilic Archaeon Sulfolobus solfataricus [Cobucci-Ponzano et al. J. Biol. Chem. (2003) 278, 14622-14631]. This enzyme, belonging to glycoside hydrolase family 29 (GH29), showed micromolar specificity for p-nitrophenyl-alpha-L-fucoside (pNp-Fuc) and promoted transfucosylation reactions by following a reaction mechanism in which the products retained the anomeric configuration of the substrate. The active site residues in GH29 enzymes are still unknown. We describe here the identification of the catalytic nucleophile of the reaction in the alpha-L-fucosidase from S. solfataricus by reactivation with sodium azide of the mutant Asp242Gly that shows a 10(3)-fold activity reduction on pNp-Fuc. The detailed stereochemical analysis of the fucosyl-azide produced by the mutant reactivated on pNp-Fuc revealed its inverted (beta-fucosyl azide) configuration compared with the substrate. This allows for the first time the unambiguous assignment of Asp242, and its homologous residues, as the nucleophilic catalytic residues of GH29 alpha-L-fucosidases. This is the first time that this approach is used for alpha-L-glycosidases, widening the applicability of this method. PMID- 12911296 TI - Solution structure of Apo Cu,Zn superoxide dismutase: role of metal ions in protein folding. AB - The solution structure of the demetalated copper, zinc superoxide dismutase is obtained for the monomeric Glu133Gln/Phe50Glu/Gly51Glu mutant through NMR spectroscopy. The demetalated protein still has a well-defined tertiary structure; however, two beta-strands containing two copper ligands (His46 and His48, beta4) and one zinc ligand (Asp83, beta5) are shortened, and the sheet formed by these strands and strands beta7 and beta8 moves away from the other strands of the beta-barrel to form an open clam with respect to a closed conformation in the holoprotein. Furthermore, loop IV which contains three zinc ligands (His63, His71, and His80) and loop VII which contributes to the definition of the active cavity channel are severely disordered, and experience extensive mobility as it results from thorough (15)N relaxation measurements. These structural and mobility data, if compared with those of the copper-depleted protein and holoprotein, point out the role of each metal ion in the protein folding, leading to the final tertiary structure of the holoprotein, and provide hints for the mechanisms of metal delivery by metal chaperones. PMID- 12911297 TI - Crystal structure of the leadzyme at 1.8 A resolution: metal ion binding and the implications for catalytic mechanism and allo site ion regulation. AB - The leadzyme is a small ribozyme, derived from in vitro selection, which catalyzes site specific, Pb(2+)-dependent RNA cleavage. Pb(2+) is required for activity; Mg(2+) inhibits activity, while many divalent and trivalent ions enhance it. The leadzyme structure consists of an RNA duplex interrupted by a trinucleotide bulge. Here, crystal structures determined to 1.8 A resolution, both with Mg(2+) as the sole divalent counterion and with Mg(2+) and Sr(2+) (which mimics Pb(2+) with respect to binding but not catalysis), reveal the metal ion interactions with both the ground state and precatalytic conformations of the leadzyme. Mg(H(2)O)(6)(2+) ions bridge complementary strands of the duplex at multiple locations by binding tandem purines of one RNA strand in the major groove. At one site, Mg(H(2)O)(6)(2+) ligates the phosphodiester backbone of the trinucleotide bulge in the ground state conformation, but not in the precatalytic conformation, suggesting (a) Mg(2+) may inhibit leadzyme activity by stabilizing the ground state and (b) metal ions which displace Mg(2+) from this site may activate the leadzyme. Binding of Sr(2+) to the presumed catalytic Pb(2+) site in the precatalytic leadzyme induces local structural changes in a manner that would facilitate alignment of the catalytic ribose 2'-hydroxyl with the scissile bond for cleavage. These data support a model wherein binding of a catalytic ion to a precatalytic conformation of the leadzyme, in conjunction with the flexibility of the trinucleotide bulge, may facilitate structural rearrangements around the scissle phosphodiester bond favoring configurations that allow bond cleavage. PMID- 12911295 TI - Effect of hydroxylation and N187-linked glycosylation on molecular and functional properties of recombinant human surfactant protein A. AB - The objective of this study was to determine the effects of proline hydroxylation in the collagen-like domain and Asn(187)-linked glycosylation in the globular domain on the molecular and functional properties of human surfactant protein A1 (SP-A1). To address this issue, SP-A1 was in vitro expressed in insect and mammalian cells. Insect cells lack prolyl 4-hydroxylase activity. A glycosylation deficient mutant SP-A1 was expressed in insect cells. In this report we present evidence that hydroxylation increased the T(m) of the collagen-like domain by 9 degrees C. Proline hydroxylation affected both the arrangement of disulfide bonding and the extent of oligomerization but did not affect conformational changes in the globular domain identified by intrinsic fluorescence. Both self association and lipid-related functions of SP-A were clearly correlated with the thermal stability of the collagen domain and the degree of oligomerization. Structural properties and lipid-related characteristics of SP-A1 expressed in mammalian cells but not in insect cells were close to that of natural human SP-A. On the other hand, the lack of glycosylation did not affect either collagen domain stability or conformational changes induced by calcium in the globular domain. However, the lack of glycosylation favored nonspecific thermally induced aggregation of the protein. PMID- 12911299 TI - Nucleotide-induced conformational changes in the Saccharomyces cerevisiae SR protein kinase, Sky1p, revealed by X-ray crystallography. AB - Conformational changes are thought to play a key role in the function of active protein kinases, although little is known about how these changes relate to the mechanism of phosphorylation. Here we present four high-resolution structures of a single crystal form of Sky1p, a constitutively active serine kinase implicated in yeast RNA processing, each in a different state of nucleotide binding. By comparing the apoenzyme structure to the ADP- and ATP-bound Sky1p structures, we have revealed conformational changes caused by ATP binding or conversion from nucleotide reactant to product. Rotation of the small lobe of the kinase closes the cleft upon binding, allowing the nucleotide to interact with residues from both lobes of the kinase, although some interactions thought to be important for phosphotransfer are missing in the ATP-containing structure. In the apoenzyme, a kinase-conserved phosphate-anchoring loop is in a twisted conformation that is incompatible with ADP and ATP binding, providing a potential mechanism for facilitating ADP release in Sky1p. The nonhydrolyzable ATP analogue AMP-PNP binds in a unique mode that fails to induce lobe closure. This observation, along with comparisons between the two independent molecules in the asymmetric unit of each structure, has provided new molecular details about how the nucleotide binds and induces closure. Finally, we have used mutational analysis to establish the importance of a glycine within the linker that connects the two lobes of Sky1p. PMID- 12911298 TI - Solution structure of the lyase domain of human DNA polymerase lambda. AB - DNA polymerase lambda (pol lambda) is a recently discovered nuclear enzyme belonging to the pol X family of DNA polymerases that exhibits a 32% sequence identity with the nuclear DNA repair protein, pol beta. Structural modeling suggests that pol lambda contains the palm, fingers, thumb, and 8 kDa lyase domains present in pol beta, as well as an additional N-terminal BRCT domain and a serine-proline-rich linker that are presumably involved in protein-protein interactions. The 8 kDa domain of pol beta is important for DNA binding and contains the dRP lyase activity, which is the rate-limiting step in the single nucleotide base excision repair (BER) pathway of damaged DNA. Recently, it was shown that the 8 kDa domain of pol lambda also contains the dRP lyase activity. To gain further insight into the catalytic mechanism of dRP removal by pol lambda, we have determined the solution structure of the 8 kDa lyase domain of human DNA pol lambda via multidimensional NMR methods and the ARIA program. The resulting structures exhibited a high degree of similarity with the 8 kDa lyase domain of pol beta. Specifically, the side chains of residues W274, R275, Y279, K307, R308, and K312 are in similar positions to the functionally important side chains of residues H34, K35, Y39, K60, K68, and K72 in the 8 kDa lyase domain of pol beta. This suggests that, on the basis of the proposed roles of these residues in pol beta, the corresponding pol lambda side chains may be involved in DNA binding and dRP lyase activity. The structural alignment of W274 (pol lambda) with H34 (pol beta) indicates that the former is probably involved in a similar base stacking interaction with template DNA at the position of the gap, in contrast with several previous proposals which aligned D272 with H34. In a few cases for which there is a nonconservative substitution in the sequence alignment, a structural comparison shows a positionally and, hence, probably a functionally equivalent residue, e.g., K60 in pol beta and K307 in pol lambda. Additionally, on the basis of the structural alignment obtained, several previously proposed mechanistic hypotheses can be evaluated. PMID- 12911300 TI - Effect of sequence on the conformation of DNA holliday junctions. AB - Structures of the DNA sequences d(CCGGCGCCGG) and d(CCAGTACbr(5)UGG) are presented here as four-way Holliday junctions in their compact stacked-X forms, with antiparallel alignment of the DNA strands. Thus, the ACC-trinucleotide motif, previously identified as important for stabilizing the junction, is now extended to PuCPy, where Pu is either an adenine or guanine, and Py is either a cytosine, 5-methylcytosine, or 5-bromouracil but not thymine nucleotide. We see that both sequence and base substituents affect the geometry of the junction in terms of the interduplex angle as well as the previously defined conformational variables, J(roll) (the rotation of the stacked duplexes about their respective helical axis) and J(slide) (the translational displacement of the stacked duplexes along their respective helical axis). The structures of the GCC and parent ACC containing junctions fall into a distinct conformational class that is relatively undistorted in terms of J(slide) and J(roll), with interduplex angles of 40-43 degrees. The substituted ACbr(5)U structure, however, is more akin to that of the distorted methylated ACm(5)C containing junction, with J(slide) (>or=2.3 A) and a similar J(roll) (164 degrees) opening the major groove-side of the junction, but shows a reduced interduplex angle. In contrast, the analogous d(CCAGTACTGG) sequence has to date been crystallized only as resolved B-DNA duplexes. This suggests that there is an electronic effect of substituents at the pyrimidine Py position on the stability of four-stranded junctions. The single crystal structures presented here, therefore, show how sequence affects the detailed geometry, and subsequently, the associated stability and conformational dynamics of the Holliday junction. PMID- 12911301 TI - Adaptation of an orthogonal archaeal leucyl-tRNA and synthetase pair for four base, amber, and opal suppression. AB - Recently, it has been shown that an amber suppressor tRNA/aminoacyl-tRNA synthetase pair derived from the tyrosyl-tRNA synthetase of Methanococcus jannaschii can be used to genetically encode unnatural amino acids in response to the amber nonsense codon, TAG. However, we have been unable to modify this pair to decode either the opal nonsense codon, TGA, or the four-base codon, AGGA, limiting us to a 21 amino acid code. To overcome this limitation, we have adapted a leucyl-tRNA synthetase from Methanobacterium thermoautotrophicum and leucyl tRNA derived from Halobacterium sp. NRC-1 as an orthogonal tRNA-synthetase pair in Escherichia coli to decode amber (TAG), opal (TGA), and four-base (AGGA) codons. To improve the efficiency and selectivity of the suppressor tRNA, extensive mutagenesis was performed on the anticodon loop and acceptor stem. The two most significant criteria required for an efficient amber orthogonal suppressor tRNA are a CU(X)XXXAA anticodon loop and the lack of noncanonical or mismatched base pairs in the stem regions. These changes afford only weak suppression of TGA and AGGA. However, this information together with an analysis of sequence similarity of multiple native archaeal tRNA sequences led to efficient, orthogonal suppressors of opal codons and the four-base codon, AGGA. Ultimately, it should be possible to use these additional orthogonal pairs to genetically incorporate multiple unnatural amino acids into proteins. PMID- 12911302 TI - Structure, stability, and function of hDim1 investigated by NMR, circular dichroism, and mutational analysis. AB - The 142 amino acid Dim1p protein is a component of the U4/U6.U5 tri-snRNP complex required for pre-mRNA splicing and interacts with multiple splicing-associated proteins. To gain further insight into the structural basis of its function, we determined the solution structure of the reduced form of the dominant negative human hDim1 (hDim1(1)(-)(128)) using multidimensional NMR spectroscopy. This dominant negative hDim1 assumes a thioredoxin-like fold, confirming previous NMR and crystallographic results. However, in contrast to a recent crystal structure, the NMR solution structure for the reduced form of hDim1(1)(-)(128) presented here, along with thermodynamic data, indicates that the presence of a disulfide bond between Cys38 and Cys79 is structurally and functionally unimportant. Comparison of the truncated hDim1(1)(-)(128) with the full-length protein, using NMR and circular dichroism spectroscopy, indicates that the 14 C-terminal residues can undergo a local unfolding transition and assume alternative conformations, which appear to play a functional role. Other residues essential for hDim1 function are identified by using mutational and genetic approaches. The residues thus identified are not identical with those previously shown to govern Dim1 interaction with defined protein partners. PMID- 12911303 TI - Structural changes of water molecules during the photoactivation processes in bovine rhodopsin. AB - Internal water molecules of rhodopsins play an important role in stabilizing the crucial ion pair comprised by the protonated retinal Schiff base and its counterion. Previous low-temperature FTIR spectroscopy of archaeal rhodopsins observed water O-D stretching vibrations at 2400-2100 cm(-1) in D(2)O, corresponding to strong hydrogen bonds. Since a water molecule bridges the protonated Schiff base and an aspartate in archaeal rhodopsins, the observed water molecules presumably hydrate the negative charges in the Schiff base region. In contrast, the FTIR spectroscopy data of bovine rhodopsin presented here revealed that there are no spectral changes of water molecules under strongly hydrogen-bonding conditions (in the range <2400 cm(-1) for O-D stretch) during the photoactivation processes. The only observed water bands were located in the >2500 cm(-1) region that corresponds to weak hydrogen bonding. These results imply that the ion pair state in vertebrate visual rhodopsins is stabilized in a manner different from that in archaeal rhodopsins. In addition, the internal water molecules that hydrate the negative charges do not play important role in the photoactivation processes of rhodopsin that involve proton transfer from the Schiff base to Glu113 upon formation of Meta II. Structural changes of the H-D exchangeable peptide amide of a beta-sheet are observed upon formation of metarhodopsin II, suggesting that motion of a beta-sheet is coupled to the proton transfer reaction from the Schiff base to its counterion. PMID- 12911304 TI - Mechanism of proton transfer inhibition by Cd(2+) binding to bacterial reaction centers: determination of the pK(A) of functionally important histidine residues. AB - The bacterial photosynthetic reaction center (RC) uses light energy to catalyze the reduction of a bound quinone molecule Q(B) to quinol Q(B)H(2). In RCs from Rhodobacter sphaeroides the protons involved in this process come from the cytoplasm and travel through pathways that involve His-H126 and His-H128 located near the proton entry point. In this study, we measured the pH dependence from 4.5 to 8.5 of the binding of the proton transfer inhibitor Cd(2+), which ligates to these surface His in the RC and inhibits proton-coupled electron transfer. At pH <6, the negative slope of the logarithm of the dissociation constant, K(D), versus pH approaches 2, indicating that, upon binding of Cd(2+), two protons are displaced; i.e., the binding is electrostatically compensated. At pH >7, K(D) becomes essentially independent of pH. A theoretical fit to the data over the entire pH range required two protons with pK(A) values of 6.8 and 6.3 (+/-0.5). To assess the contribution of His-H126 and His-H128 to the observed pH dependence, K(D) was measured in mutant RCs that lack the imidazole group of His H126 or His-H128 (His --> Ala). In both mutant RCs, K(D) was approximately pH independent, showing that Cd(2+) does not displace protons upon binding in the mutant RCs, in contrast to the native RC in which His-H126 and His-H128 are the predominant contributors to the observed pH dependence of K(D). Thus, Cd(2+) inhibits RC function by binding to functionally important histidines. PMID- 12911305 TI - Structural dynamics of myoglobin: spectroscopic and structural characterization of ligand docking sites in myoglobin mutant L29W. AB - We have studied CO binding to the heme and CO migration among protein internal cavities after photodissociation in sperm whale carbonmonoxy myoglobin (MbCO) mutant L29W using Fourier transform infrared (FTIR) spectroscopy combined with temperature derivative spectroscopy (TDS) and kinetic experiments at cryogenic temperatures. Photoproduct intermediates, characterized by CO at particular locations in the protein, were selectively enhanced by applying special laser illumination protocols. These studies were performed on the L29W mutant protein and a series of double mutants constructed so that bulky amino acid side chains block passageways between cavities or fill these sites. Binding of xenon was also employed as an alternative means of occluding cavities. All mutants exhibit two conformations, A(I) and A(II), with distinctly different photoproduct states and ligand binding properties. These differences arise mainly from different positions of the W29 and H64 side chains in the distal heme pocket [Ostermann, A., et al. (2000) Nature 404, 205-208]. The detailed knowledge of the interplay between protein structure, protein dynamics, and ligand migration at cryogenic temperatures allowed us to develop a dynamic model that explains the slow CO and O(2) bimolecular association observed after flash photolysis at ambient temperature. PMID- 12911307 TI - Barley serine proteinase inhibitor 2-derived cyclic peptides as potent and selective inhibitors of convertases PC1/3 and furin. AB - Proprotein convertases (PCs) are serine proteases containing a subtilisin-like catalytic domain that are involved in the conversion of hormone precursors into their active form. This study aims at designing small cyclic peptides that would specifically inhibit two members of this family of enzymes, namely, the neuroendocrine PC1/3 and the ubiquitously expressed furin. We studied peptide sequences related to the 18-residue loop identified as the active site of the 83 amino acid barley serine protease inhibitor 2 (BSPI-2). Peptides incorporating mutations at various positions in the sequence were synthesized on solid phase and purified by HPLC. Cyclization was achieved by the introduction of a disulfide bridge between the two Cys residues located at both the N- and C-terminal extremities. Peptides VIIA and VIIB incorporating P4Arg, P2Lys, P1Arg, and P2'Lys were the most potent inhibitors with K(i) around 4 microM for furin and around 0.5 microM for PC1/3. Whereas peptide VIIB behaved as a competitive inhibitor of furin, peptide VIIA acted as a noncompetitive one. However, all peptides were eventually cleaved after variable incubation times by PC1/3 or furin. To avoid this problem, we incorporated at the identified cleavage site a nonscissile aminomethylene bond (psi[CH(2)-NH]). Those pseudopeptides, in particular peptide VIID, were shown not to be cleaved and to inhibit potently furin. Conversely, they were not able to inhibit PC1/3 at all. Those results show the validity of this approach in designing new effective PC inhibitors showing a certain level of discrimination between PC1/3 and furin. PMID- 12911306 TI - Structural dynamics of myoglobin: effect of internal cavities on ligand migration and binding. AB - Using Fourier transform infrared (FTIR) spectroscopy combined with temperature derivative spectroscopy (TDS) at cryogenic temperatures, we have studied CO binding to the heme and CO migration among cavities in the interior of sperm whale carbonmonoxy myoglobin (MbCO) after photodissociation. Photoproduct intermediates, characterized by CO in different locations, were selectively enhanced by laser illumination at specific temperatures. Measurements were performed on the wild-type protein and a series of mutants (L104W, I107W, I28F, and I28W) in which bulky amino acid side chains were introduced to block passageways between cavities or to fill these sites. Binding of xenon was also employed as an alternative means of filling cavities. In all samples, photolyzed CO ligands were observed to initially bind at primary docking site B in the vicinity of the heme iron, from where they migrate to the secondary docking sites, the Xe4 and/or Xe1 cavities. To examine the relevance of these internal docking sites for physiological ligand binding, we have performed room temperature flash photolysis on the entire set of proteins in the CO- and O(2) bound form. Together with the cryospectroscopic results, these data provide a clear picture of the role of the internal sites for ligand escape from and binding to myoglobin. PMID- 12911308 TI - Highly nonexponential kinetics in the early-phase refolding of proteins at low temperatures. AB - Theory and simulations predict that the folding kinetics of protein-like heteropolymers become nonexponential and glassy (i.e., controlled by escape from different low-energy misfolded states) at low temperatures, but there was little experimental evidence for such behavior of proteins. We have developed a stopped flow instrument working reliably down to -40 degrees C with high mixing capability and applied it to study the refolding kinetics of horse cytochrome c (cyt c) and hen egg white lysozyme at temperatures below 0 degrees C in the presence of antifreeze NaCl, LiCl, or ethylene glycol and above 0 degrees C in the presence and absence of antifreeze. The refolding was initiated by rapid dilution of the guanidine hydrochloride unfolded proteins, and the kinetics were monitored by intrinsic tryptophan fluorescence. Highly nonexponential kinetics extended over 3 decades in time (0.01-10 s) were observed in the early phases of the refolding of cyt c and lysozyme in the temperature range of -35 to 5 degrees C. These results are in agreement with the theoretical prediction, suggesting that the folding energy landscapes of these proteins are rugged in the upper portions. PMID- 12911309 TI - Irreversible binding and activity control of the 1,2-diacylglycerol 3 glucosyltransferase from Acholeplasma laidlawii at an anionic lipid bilayer surface. AB - 1,2-Diacylglycerol 3-glucosyltransferase is associated with the membrane surface catalyzing the synthesis of the major nonbilayer-prone lipid alpha-monoglucosyl diacylglycerol (MGlcDAG) from 1,2-DAG in the cell wall-less Acholeplasma laidlawii. Phosphatidylglycerol (PG), but not neutral or zwitterionic lipids, seems to be essential for an active conformation and function of the enzyme. Surface plasmon resonance analysis was employed to study association of the enzyme with lipid bilayers. Binding kinetics could be well fitted only to a two state model, implying also a (second) conformational step. The enzyme bound less efficiently to liposomes containing only zwitterionic lipids, whereas increasing molar fractions of the anionic PG or cardiolipin (CL) strongly promoted binding by improved association (k(a1)), and especially a decreased rate of return (k(d2)) from the second state. This yielded a very low overall dissociation constant (K(D)), corresponding to an essentially irreversible membrane association. Both liposome binding and consecutive activity of the enzyme correlated with the PG concentration. The importance of the electrostatic interactions with anionic lipids was shown by quenching of both binding and activity with increasing NaCl concentrations, and corroborated in vivo for an active enzyme-green fluorescent protein hybrid in Escherichia coli. Nonbilayer prone lipids substantially enhanced enzyme-liposome binding by promoting a changed conformation (decreasing k(d2)), similar to the anionic lipids, indicating the importance of hydrophobic interactions and a curvature packing stress. For CL and the nonbilayer lipids, effects on enzyme binding and consecutive activity were not correlated, suggesting a separate lipid control of activity. Similar features were recorded with polylysine (cationic) and polyglutamate (anionic) peptides present, but here probably dependent on the selective charge interactions with the enzyme N- and C-domains, respectively. A lipid-dependent conformational change and PG association of the enzyme were verified by circular dichroism, intrinsic tryptophan, and pyrene-probe fluorescence analyses, respectively. It is concluded that an electrostatic association of the enzyme with the membrane surface is accompanied by hydrophobic interactions and a conformational change. However, specific lipids, the curvature packing stress, and proteins or small molecules bound to the enzyme can modulate the activity of the bound A. laidlawii MGlcDAG synthase. PMID- 12911310 TI - Refolding of amphioxus insulin-like peptide: implications of a bifurcating evolution of the different folding behavior of insulin and insulin-like growth factor 1. AB - Insulin and insulin-like growth factor 1 (IGF-1) share high sequence homology, but their folding behaviors are significantly different: insulin folds into one unique thermodynamically controlled structure, while IGF-1 folds into two thermodynamically controlled disulfide isomers. However, the origin of their different folding behaviors is still elusive. The amphioxus insulin-like peptide (ILP) is thought to be the common ancestor of insulin and IGF-1. A recombinant single-chain ILP has been expressed previously, and now its folding behavior is investigated. The folding behavior of ILP shows the characteristics of both insulin and IGF-1. On one hand, two thermodynamically controlled disulfide isomers of ILP have been identified; on the other hand, the content of isomer 1 (its disulfides are deduced identical to those of swap IGF-1) is much less than that of isomer 2 (its disulfides are deduced identical to those of native IGF-1); that is, more than 96% of ILP folds into the native structure. The present results suggest that the different folding behaviors of insulin and IGF-1 are acquired through a bifurcating evolution: the tendency of forming the thermodynamically controlled non-native disulfide isomer is diminished during evolution from ILP to insulin, while this tendency is amplified during evolution from ILP to IGF-1. Moreover, the N-terminal Gln residue of ILP can spontaneously form a pyroglutamate residue, and its cyclization has a significant effect on the folding behavior of ILP: the percentage of isomer 1 is approximately 2-fold that of isomer 1 of the noncyclized ILP; that is, isomer 1 becomes more favored when the N-terminal residue of ILP is cyclized. So, we deduce that the N-terminal residues have a significant effect on the folding properties of insulin, IGF-1, and ILP. PMID- 12911311 TI - Rho-specific Bacillus cereus ADP-ribosyltransferase C3cer cloning and characterization. AB - C3-like ADP-ribosyltransferases represent an expanding family of related exoenzymes, which are produced by Clostridia and various Staphylococcus aureus strains. Here we report on the cloning and biochemical characterization of an ADP ribosyltransferase from Bacillus cereus strain 2339. The transferase encompasses 219 amino acids; it has a predicted mass of 25.2 kDa and a theoretical isoelectric point of 9.3. To indicate the relationship to the family of C3-like ADP-ribosyltransferases, we termed the enzyme C3cer. The amino acid sequence of C3cer is 30 to 40% identical to other C3-like exoenzymes. By site-directed mutagenesis, Arg(59), Arg(97), Tyr(151), Arg(155), Thr(178), Tyr(180), Gln(183), and Glu(185) of recombinant C3cer were identified as pivotal residues of enzyme activity and/or protein substrate recognition. Precipitation experiments with immobilized RhoA revealed that C3cerTyr(180), which is located in the so-called "ADP-ribosylating toxin turn-turn" (ARTT) motif, plays a major role in the recognition of RhoA. Like other C3-like exoenzymes, C3cer ADP-ribosylates preferentially RhoA and RhoB and to a much lesser extent RhoC. Because the cellular accessibility of recombinant C3cer is low, a fusion toxin (C2IN-C3cer), consisting of the N-terminal 225 amino acid residues of the enzyme component of C2 toxin from Clostridium botulinum and C3cer was used to study the cytotoxic effects of the transferase. This fusion toxin caused rounding up of Vero cells comparable to the effects of Rho-inactivating toxins. PMID- 12911312 TI - Mass spectrometric identification of N- and O-glycosylation sites of full-length rat selenoprotein P and determination of selenide-sulfide and disulfide linkages in the shortest isoform. AB - Rat selenoprotein P is an extracellular glycoprotein of 366 amino acid residues that is rich in cysteine and selenocysteine. Plasma contains four isoforms that differ principally by length at the C-terminal end. Mass spectrometry was used to identify sites of glycosylation on the full-length protein. Of the potential N glycosylation sites, three located at residues 64, 155, and 169 were occupied, while the two at residues 351 and 356 were not occupied. Threonine 346 was variably O-glycosylated. Thus, full-length selenoprotein P is both N- and O glycosylated. The shortest isoform of selenoprotein P, which terminates at residue 244, was analyzed for selenide-sulfide and disulfide linkages. In this isoform, a single selenocysteine and seven cysteines are present. Mass spectrometric analysis indicated that a selenide-sulfide bond exists between Sec40 and Cys43. Two disulfides were also detected as Cys149-Cys167 and Cys153 Cys156. The finding of a selenide-sulfide bond in the shortest isoform is compatible with a redox function of this pair that might be analogous to the selenol-thiol pair near the C terminus of animal thioredoxin reductase. The disulfide formed by Cys153-Cys156 also has some characteristics of a redox active pair. PMID- 12911313 TI - Ascaris suum NAD-malic enzyme is activated by L-malate and fumarate binding to separate allosteric sites. AB - The kinetic mechanism of activation of the mitochondrial NAD-malic enzyme from the parasitic roundworm Ascaris suum has been studied using a steady-state kinetic approach. The following conclusions are suggested. First, malate and fumarate increase the activity of the enzyme in both reaction directions as a result of binding to separate allosteric sites, i.e., sites that exist in addition to the active site. The binding of malate and fumarate is synergistic with the K(act) decreasing by >or=10-fold at saturating concentrations of the other activator. Second, the presence of the activators decreases the K(m) for pyruvate 3-4-fold, and the K(i) (Mn) >or=20-fold in the direction of reductive carboxylation; similar effects are obtained with fumarate in the direction of oxidative decarboxylation. The greatest effect of the activators is thus expressed at low reactant concentrations, i.e., physiologic concentrations of reactant, where activation of >or=15-fold is observed. A recent crystallographic structure of the human mitochondrial NAD malic enzyme [13] shows fumarate bound to an allosteric site. Site-directed mutagenesis was used to change R105, homologous to R91 in the fumarate activator site of the human enzyme, to alanine. The R105A mutant enzyme exhibits the same maximum rate and V/K(NAD) as does the wild-type enzyme, but 7-8-fold decrease in both V/K(malate) and V/K(Mg), indicating the importance of this residue in the activator site. In addition, neither fumarate nor malate activates the enzyme in either reaction direction. Finally, a change in K143 (a residue in a positive pocket adjacent to that which contains R105), to alanine results in an increase in the K(act) for malate by about an order of magnitude such that it is now of the same magnitude as the K(m) for malate. The K143A mutant enzyme also exhibits an increase in the K(act) for fumarate (in the absence of malate) from 200 microM to about 25 mM. PMID- 12911314 TI - Oxidase domains in epothilone and bleomycin biosynthesis: thiazoline to thiazole oxidation during chain elongation. AB - The natural products epothilone and bleomycin are assembled by hybrid polyketide/nonribosomal peptide synthetases. Of note in these assembly lines is the conversion of internal cysteine residues into thiazolines and their subsequent oxidation to heteroaromatic thiazole rings. We have excised the EpoB oxidase domain, EpoB-Ox, proposed to be responsible for thiazoline to thiazole oxidation in epothilone biosynthesis, and expressed it in soluble form in Escherichia coli. The purified domain is an FMN-containing flavoprotein that demonstrates thiazoline to thiazole oxidase activity when incubated with thioester substrate mimics. Kinetic parameters were determined for both thiazoline and oxazoline substrates, with k(cat) values ranging between 48.8 and 0.55 min(-1). While the physiological electron acceptor is not yet known, molecular oxygen is needed in these in vitro assays to mediate reoxidation of reduced FMN. Additionally, the oxidase domain-containing BlmIII from the bleomycin assembly line was heterologously expressed and purified. BlmIII is also an FMN-containing protein with activity similar to EpoB-Ox. This work marks the first direct characterization of nonribosomal peptide synthetase oxidase domain activity and will lead to further exploration of these flavoproteins. PMID- 12911315 TI - BlmIII and BlmIV nonribosomal peptide synthetase-catalyzed biosynthesis of the bleomycin bithiazole moiety involving both in cis and in trans aminoacylation. AB - Cloning and sequence analysis of the bleomycin (BLM) biosynthetic gene cluster predicted that the two nonribosomal peptide synthetases (NRPSs), BlmIV and BlmIII, are responsible for the biosynthesis of the BLM bithiazole moiety. BlmIV is a seven domain (C(2)-A(2)-PCP(2)-Cy(1)-A(1)-PCP(1)-Cy(0)) NRPS, and BlmIII is a three domain (A(0)-PCP(0)-Ox) NRPS. The three domains of Cy(1)-A(1)-PCP(1) residing on the BlmIV subunit, the four domains of Cy(0) residing on the BlmIV subunit, and A(0)-PCP(0)-Ox residing on the BlmIII subunit constitute the two thiazole-forming NRPS-1 and NRPS-0 modules, respectively. BlmIII-A(0) was predicted to be nonfunctional, raising the question of how the NRPS-0 module activates and loads the Cys substrate to its cognate BlmIII-PCP(0). The NRPS-0 module consists of domains residing on two different subunits, requiring precise protein-protein interaction. Here, we report the production of the BlmIV and BlmIII NRPSs as an excised domain(s), module, or intact subunit form and biochemical characterizations of the resultant enzymes in vitro for their roles in BLM bithiazole biosynthesis. Our results (a) confirm that BlmIII-A(0) is a naturally occurring nonfunctional mutant, (b) demonstrate that BlmIV-A(1) activates Cys and catalyzes both in cis aminoacylation of BlmIV-PCP(1) (for NRPS 1) and in trans aminoacylation of BlmIII-PCP(0) (for NRPS-0), and (c) reveal that the C-terminus of the BlmIV subunit, characterized by the unprecedented AGHDDD(G) and PGHDDG repeats, is absolutely required for in trans aminoacylation of BlmIII PCP(0). These findings underscore the flexibility and versatility of NRPSs in both structure and mechanism for natural product biosynthesis and provide an outstanding opportunity to study the molecular recognition and protein-protein interaction mechanism in NRPS assembly line enzymology. PMID- 12911316 TI - Kinetic studies of protein farnesyltransferase mutants establish active substrate conformation. AB - The zinc metalloenzyme protein farnesyltransferase (FTase) catalyzes the transfer of a 15-carbon farnesyl moiety from farnesyl diphosphate (FPP) to a cysteine residue near the C-terminus of a protein substrate. Several crystal structures of inactive FTase.FPP.peptide complexes indicate that K164alpha interacts with the alpha-phosphate and that H248beta and Y300beta form hydrogen bonds with the beta phosphate of FPP [Strickland, C. L., et al. (1998) Biochemistry 37, 16601-16611]. Mutations K164Aalpha, H248Abeta, and Y300Fbeta were prepared and analyzed by single turnover kinetics and ligand binding studies. These mutations do not significantly affect the enzyme affinity for FPP but do decrease the farnesylation rate constant by 30-, 10-, and 500-fold, respectively. These mutations have little effect on the pH and magnesium dependence of the farnesylation rate constant, demonstrating that the side chains of K164alpha, Y300beta, and H248beta do not function either as general acid-base catalysts or as magnesium ligands. Mutation of H248beta and Y300beta, but not K164alpha, decreases the farnesylation rate constant using farnesyl monophosphate (FMP). These data suggest that, contrary to the conclusions derived from analysis of the static crystal structures, the transition state for farnesylation is stabilized by interactions between the alpha-phosphate of the isoprenoid substrate and the side chains of Y300beta and H248beta. These results suggest an active substrate conformation for FTase wherein the C1 carbon of the FPP substrate moves toward the zinc-bound thiolate of the protein substrate to react, resulting in a rearrangement of the diphosphate group relative to its ground state position in the binding pocket. PMID- 12911317 TI - Analysis of bisdioxopiperazine dexrazoxane binding to human DNA topoisomerase II alpha: decreased binding as a mechanism of drug resistance. AB - Topoisomerase II is an ATP-operated clamp that effects topological changes by capturing a double stranded DNA segment and transporting it through another DNA molecule. Despite the extensive use of topoisomerase II-targeted drugs in cancer chemotherapy and the impact of drug resistance on the efficacy of treatment, much remains unknown concerning the interactions between these agents and topoisomerase II. To identify the interaction of the bisdioxopiperazine dexrazoxane (ICRF-187) with topoisomerase II, we developed a rapid gel-filtration assay and characterized the binding of ((3)H)-dexrazoxane to human topoisomerase II alpha. Dexrazoxane binds to human topoisomerase II alpha in the presence of DNA and ATP with an apparent K(d) of 23 microM and a stoichiometry of 1 drug molecule per enzyme dimer. Various N-terminal single amino acid substitutions in human topoisomerase II alpha that were previously shown to confer specific bisdioxopiperazine resistance either totally abolished drug binding or resulted in less efficient binding. The effect of the various mutations on drug binding correlated well with their effect on drug resistance in vivo and in vitro. Interestingly, an altered active site tyrosine mutant of human topoisomerase II alpha, which is incapable of carrying out DNA strand passage, was unable to bind dexrazoxane, which agrees with the drug's proposed mechanism of action late in the topoisomerase II catalytic cycle. The direct correlation between the level of drug binding and dexrazoxane resistance is consistent with a decreased drug binding mechanism of action for these dexrazoxane resistance conferring mutations. PMID- 12911318 TI - Repair of DNA containing Fapy.dG and its beta-C-nucleoside analogue by formamidopyrimidine DNA glycosylase and MutY. AB - Fapy.dG is produced in DNA as a result of oxidative stress. Under some conditions Fapy.dG is formed in greater yields than 8-oxodG from a common chemical precursor. Recently, Fapy.dG and its C-nucleoside analogue were incorporated in chemically synthesized oligonucleotides at defined sites. Like 8-oxodG, Fapy.dG instructs DNA polymerase to misincorporate dA opposite it in vitro. The interactions of DNA containing Fapy.dG or the nonhydrolyzable analogue with Fpg and MutY are described. Fpg excises Fapy.dG (K(M) = 2.0 nM, k(cat) = 0.14 min( 1)) opposite dC approximately 17-fold more efficiently than when mispaired with dA, which is misinserted by DNA polymerase in vitro. Fpg also prefers to bind duplexes containing Fapy.dG.dC or beta-C-Fapy.dG.dC compared to those in which the lesion is opposite dA. MutY incises dA when it is opposite Fapy.dG and strongly binds duplexes containing the lesion or beta-C-Fapy.dG. Incision from Fapy.dG.dA is faster than from dG.dA mispairs but slower than from DNA containing 8-oxodG opposite dA. These data demonstrate that Fapy.dG closely resembles the interactions of 8-oxodG with two members of the GO repair pathway in vitro. The similar effects of Fapy.dG and 8-oxodG on DNA polymerase and repair enzymes in vitro raise the question as to whether Fapy.dG elicits similar effects in vivo. PMID- 12911320 TI - CofE catalyzes the addition of two glutamates to F420-0 in F420 coenzyme biosynthesis in Methanococcus jannaschii. AB - The protein product of the Methanococcus jannaschii MJ0768 gene has been expressed in Escherichia coli, purified to homogeneity, and shown to catalyze the GTP-dependent addition of two l-glutamates to the l-lactyl phosphodiester of 7,8 didemethyl-8-hydroxy-5-deazariboflavin (F(420)-0) to form F(420)-0-glutamyl glutamate (F(420)-2). Since the reaction is the fifth step in the biosynthesis of coenzyme F(420), the enzyme has been designated as CofE, the product of the cofE gene. Gel filtration chromatography indicates CofE is a dimer. The enzyme has no recognized sequence similarity to any previously characterized proteins. The enzyme has an absolute requirement for a divalent metal ion and a monovalent cation. Among the metal ions tested, a mixture of Mn(2+), Mg(2+), and K(+) is the most effective. CofE catalyzes amide bond formation with the cleavage of GTP to GDP and inorganic phosphate, likely involving the activation of the free carboxylate group of F(420)-0 to give an acyl phosphate intermediate. Evidence for the occurrence of this intermediate is presented. A reaction mechanism for the enzyme is proposed and compared with other members of the ADP-forming amide bond ligase family. PMID- 12911321 TI - Biosynthesis of docosahexaenoic acid in Euglena gracilis: biochemical and molecular evidence for the involvement of a Delta4-fatty acyl group desaturase. AB - Docosahexaenoic acid (DHA) can be synthesized via alternative routes from which only the omega3/omega6-pathways involve the action of a Delta4-fatty acid desaturase. We examined the suitability of Euglena gracilis, Thraustochytrium sp., Schizochytrium sp., and Crypthecodinium cohnii to serve as sources for cloning a cDNA encoding a Delta4-fatty acid desaturase. For this purpose we carried out in vivo labeling studies with radiolabeled C22 polyunsaturated fatty acid substrates. Schizochytrium sp. was unable to convert exogenously supplied [2 (14)C]-docosapentaenoic acid (DPA, 22:5(Delta)(7,10,13,16,19)) to DHA, while E. gracilis and Thraustochytrium sp. carried out this desaturation very efficiently. Hydrogenation and alpha-oxidation of the labeled DHA isolated from these two organisms showed that it was the result of direct Delta4-desaturation and not of substrate breakdown and resynthesis. To clone the desaturase gene, a cDNA library of E. gracilis was subjected to mass sequencing. A full-length clone with highest homology to the Delta4-desaturase of Thraustochytrium sp. was isolated, and its function was verified by heterologous expression in yeast. The desaturase efficiently converted DPA to DHA. Analysis of the substrate specificity demonstrated that the enzyme activity was not limited to C22 fatty acids, since it also efficiently desaturated C16 fatty acids. The enzyme showed strict Delta4 regioselectivity and required the presence of a Delta7-double bond in the substrate. Positional analysis of phosphatidylcholine revealed that the proportion of the Delta4-desaturated products was up to 20 times higher in the sn 2 position than in the sn-1 position. PMID- 12911319 TI - Oxidized guanine lesions as modulators of gene transcription. Altered p50 binding affinity and repair shielding by 7,8-dihydro-8-oxo-2'-deoxyguanosine lesions in the NF-kappaB promoter element. AB - A number of common promoter elements that drive transcription of redox sensitive genes have runs of guanines in their transcription factor recognition sequence. A paradox exists insomuch that the same guanine runs necessary for transcription factor recognition are thermodynamically prone to oxidative modification, potentially altering the binding affinity of transcription factors. 7,8-Dihydro-8 oxo-2'-deoxyguanosine (8-oxo-dG) is a common oxidative modification of guanine that is generated by a variety of metals and reactive oxygen species. We have used the p50 subunit of the NF-kappaB transcription factor to show that oxidation of guanine to 8-oxo-dG at sites critical for protein recognition impacts transcription factor binding affinity differently depending upon the site of oxidation. It can be argued that the impact of such oxidation will be minimal in repair proficient cells. Therefore, we have developed an assay to assess the ability of these lesions to be shielded by transcription factor binding from recognition and repair by base excision repair (BER) enzymes. In this study, 8 oxo-dG was substituted for guanine at sites G(1)-G(4) in the NF-kappaB sequence 5'-d(AGTTGAG(1)G(2)G(3)G(4)ACTTTCCCAGCC)-3'. We have observed that substitution of 8-oxo-dG at the G(1) site increases p50 binding affinity by approximately 2.5 fold compared to that of the unmodified DNA sequence, while substitution at G(3) reduces the binding affinity by approximately 4-fold. Substitution of 8-oxo-dG at the G(2) and G(4) sites had a minimal impact on p50 binding affinity. Both Escherichia coli fapy glycosylase (Fpg) and human 8-oxo-DNA glycosylase (hOGG1) recognized and cleaved 8-oxo-dG at all four sites within the promoter element. The addition of the p50 transcription factor shielded these lesions from cleavage by the glycosylase in a manner that correlated with the binding affinities of p50 for the different modified sites. These data imply that lesion formation in DNA response elements can modulate gene transcription during oxidative events and that protein binding to these modified sites may allow these lesions to persist on a time scale that impacts global cellular gene transcription. PMID- 12911322 TI - Accessibility of cysteine residues in a cytoplasmic loop of CitS of Klebsiella pneumoniae is controlled by the catalytic state of the transporter. AB - The citrate transporter CitS of Klebsiella pneumoniae is a secondary transporter that transports citrate in symport with two sodium ions and one proton. Treatment of CitS with the alkylating agent N-ethylmaleimide resulted in a complete loss of transport activity. Treatment of mutant proteins in which the five endogenous cysteine residues were mutated into serines in different combinations revealed that two cysteine residues located in the C-terminal cytoplasmic loop, Cys-398 and Cys-414, were responsible for the inactivation. Labeling with the membrane impermeable methanethiosulfonate derivatives MTSET and MTSES in right-side-out membrane vesicles showed that the cytoplasmic loop was accessible from the periplasmic side of the membrane. The membrane impermeable but more bulky maleimide AmdiS did not inactivate the transporter in right-side-out membrane vesicles. Inactivation by N-ethylmaleimide, MTSES, and MTSET was prevented by the presence of the co-ion Na(+). Protection was obtained upon binding 2 Na(+), which equals the transport stoichiometry. In the absence of Na(+), the substrate citrate had no effect on the inactivation by permeable or impermeable thiol reagents. In contrast, when subsaturating concentrations of Na(+) were present, citrate significantly reduced inactivation suggesting ordered binding of the substrate and co-ion; citrate is bound after Na(+). In the presence of the proton motive force, the reactivity of the Cys residues was increased significantly for the membrane permeable N-ethylmaleimide, while no difference was observed for the membrane impermeable thiol reagents. The results are discussed in the context of a model for the opening and closing of the translocation pore during turnover of the transporter. PMID- 12911324 TI - Interaction of amoebapores and NK-lysin with symmetric phospholipid and asymmetric lipopolysaccharide/phospholipid bilayers. AB - Amoebapores from protozoan parasite Entamoeba histolytica and NK-lysin of porcine cytotoxic lymphocytes belong to the same family of saposin-like proteins. In addition to the structural similarity, amoebapores and NK-lysin are both highly effective against prokaryotic and eukaryotic target cells in that they permeabilize the target cell membranes. Here, we have investigated in detail the protein/lipid interaction for the three isoforms of amoebapore and NK-lysin. Results obtained from electrical measurements on planar bilayer membranes, including reconstitution models of the lipid matrix of the outer membrane of Escherichia coli and phospholipid membranes, fluorescence energy transfer spectroscopy with liposomes, and monolayer measurements on a Langmuir trough, provided information on lipid preferences, pH dependences, and membrane interaction mechanisms. The three amoebapores led to the formation of transient pores with similar characteristics in conductance, sublevels, and lifetime for the different isoforms. The conductance of the pores was dependent on the polarity of the applied clamp voltage, and the distribution of the sublevels was affected by the value of the clamp voltage. The size of the pores and distribution of conductance sublevels differed between symmetric phospholipid and asymmetric lipopolysaccharide/phospholipid bilayers. Notably, NK-lysin caused the formation of well-defined pores, which were lipid- and voltage-dependent, and their characteristics differed from those induced by amoebapores; e.g., the protein concentration necessary to induce pore formation was 20 times higher. The biophysical data give important information on the mode of action of these small effector proteins, which may further lead to a better understanding of peptide membrane interactions in general. PMID- 12911323 TI - Transient kinetics and mechanics of myosin's force-generating rotation in muscle: resolution of millisecond rotational transitions in the spin-labeled myosin light chain domain. AB - We have used electron paramagnetic resonance (EPR) of spin-labeled scallop muscle, in conjunction with laser flash photolysis of caged ATP, to resolve millisecond rotational transitions of the myosin light-chain domain (LCD) during transient force generation. We previously used EPR to resolve two distinct orientations of the LCD [Baker, J. E., Brust-Mascher, I., Ramachandran, S., LaConte, L. E., and Thomas, D. D. (1998) Proc. Natl. Acad. Sci. U.S.A. 95, 2944 2949], correlated these structural states with biochemical states in the actin myosin ATPase reaction, and showed that a small shift in the steady-state distribution between these two LCD orientations (i.e., a net lever arm rotation) is associated with force generation in muscle. In the study presented here, we measured millisecond changes in this orientational distribution (i.e., the rates of transition between the two LCD orientations) in muscle following flash photolysis of caged ATP, in both the presence and absence of Ca. The transient acquired in the absence of Ca is dominated by a rapid (1/tau(1) = 37 s(-1)) disordering transition from the single orientation in rigor to the bimodal orientation distribution observed for detached cross-bridges in relaxation (i.e., the reversal of the lever arm rotation), followed by a recovery phase (1/tau(2) = 2.4 s(-1)) of very small amplitude (small fraction of heads participating). In the presence of Ca, the transient exhibited a similar initial disordering phase (1/tau(1) = 38.5 s(-1)), followed by a recovery phase (1/tau(2) = 8.33 s(-1)) of substantial amplitude, corresponding to the forward rotation and ordering of the lever arm. A standard kinetic model was used to fit these data, revealing rate constants consistent with those previously determined by other methods. Surprisingly, a comparison of the EPR transients with force transients reveals that the rate of force development (91 s(-1)) is faster than the rate of the forward lever arm rotation (8 s(-1)). This observed relationship between the kinetics of the lever arm rotation and transient force development in muscle provides new insight into how myosin both generates and responds to muscle force. PMID- 12911325 TI - Structural characterization of the ADAM 16 disintegrin loop active site. AB - ADAM's have various roles in intercellular adhesion and are thought to function by binding integrins through a 13 amino acid motif called the disintegrin loop. Xenopus laevis sperm express the protein ADAM 16, and peptides with the sequence of its disintegrin loop cause downstream events in eggs that require a rise in intracellular calcium similar to that occurring at fertilization. We characterized the portion of the ADAM 16 disintegrin loop responsible for causing egg activation. A peptide based on the C-terminal half of the motif, which includes a known integrin-binding sequence, is a partial agonist of calcium release. A peptide with the N-terminal sequence of the motif activates eggs in a manner virtually identical to the full-length peptide but lacks a recognized integrin-binding sequence. None of these peptides alter the permeability or fluidity of liposomes made from membrane lipids of X. laevis eggs. This result reflects the fact that the peptides do not cause calcium to leak across the egg membrane and indirectly provides evidence that they act through a receptor on the egg surface. The infrared spectrum of the full-length peptide has a strong absorption peak corresponding to a beta-turn. We predict this structure occurs at the N-terminal sequence MPKT. A rearranged peptide lacking any turns fails to activate eggs. These results provide the first structural information about the active site of an ADAM disintegrin loop. We interpret these results in terms of active site sequences from other ADAM's and the role of integrins during fertilization. PMID- 12911326 TI - Redox labile site in a Zn4 cluster of Cu4,Zn4-metallothionein-3. AB - Human metallothionein-3 (MT-3) is a neuronal inhibitory factor mainly expressed in brain and downregulated in Alzheimer's disease. The neuroinhibitory activity has been established for native Cu(4),Zn(3)-MT-3 and recombinant Zn(7)-MT-3. However, there is only limited knowledge about the structure and properties of the former metalloform. We have now generated native-like MT-3 through direct Cu(I) and Zn(II) incorporation into the recombinant apoprotein. Its characterization revealed monomeric Cu(4),Zn(4)-MT-3 containing metal-thiolate clusters located in two mutually interacting protein domains, a Cu(4) cluster in the beta-domain and a Zn(4) cluster in the alpha-domain. Using the PC12 cell line, the nontoxic nature of the protein was demonstrated. The results of electronic absorption and Cu(I) luminescence at 77 K showed that the Cu(4) cluster possesses an unprecedented stability in air. In contrast, the Zn(4) cluster is air sensitive. Its oxidation results in the release of one Zn(II) and the formation of a Zn(3) cluster, i.e., Cu(4),Zn(3)-MT-3. This process can be prevented or reversed under reducing conditions. The determined apparent stability constant for the Zn(4) cluster of 2.4 x 10(11) M(-1) is similar to that obtained for other zinc-containing MTs. This suggests that a substantially increased nucleophilic reactivity of specific thiolate ligands is responsible for this effect. Thus, the Zn(4) cluster in MT-3 may play a redox-dependent regulatory role. PMID- 12911327 TI - Angiotensinogen gene M235T polymorphism and reduction in wall thickness in response to antihypertensive treatment. AB - The angiotensinogen M235T polymorphism has been linked to hypertension and cardiovascular disease. Carotid intima-media thickness (IMT) is an early marker of atherosclerosis. The objectives of the present study were to determine in previously untreated essential hypertensive patients whether carotid IMT was associated with the M235T polymorphism, and to determine whether the M235T polymorphism could influence the reduction of carotid IMT by antihypertensive treatment. Common carotid artery IMT was determined with a high-definition echotracking system in 98 previously untreated hypertensive patients in a cross sectional study. A subgroup of 56 patients was included in a randomized double blind parallel group study comparing the effect of the angiotensin-converting enzyme-inhibitor enalapril with that of the beta-blocker celiprolol during a 5 month period. In the cross-sectional study, a multivariate analysis showed that the M235T genotype was a significant independent determinant of carotid IMT, explaining 7% of the variance. Carotid IMT was higher in patients homozygous for the T allele than in MM patients. In the longitudinal study, the reduction in carotid IMT after antihypertensive treatment was significantly ( P <0.01) higher in patients carrying the TT genotype than in patients carrying the MM genotype, despite similar reductions in blood pressure and independently of drug type. In conclusion, these data suggest that the angiotensinogen TT genotype at position 235 is a genetic marker for early carotid atherosclerosis in a hypertensive population and its regression under antihypertensive treatment. PMID- 12911328 TI - Thimet oligopeptidase expression is differentially regulated in neuroendocrine and spermatid cell lines by transcription factor binding to SRY (sex-determining region Y), CAAT and CREB (cAMP-response-element-binding protein) promoter consensus sequences. AB - The zinc metalloprotease thimet oligopeptidase (EP24.15) is found predominantly in the neuroendocrine-gonadal axis where it is implicated in the processing of bioactive peptides, including GnRH (gonadotropin-releasing hormone), beta neoendorphin, alpha-neoendorphin and dynorphin(1-8), the progression of spermatogenesis and the normal clearance of beta-amyloid in brain cells. Regulation of the enzyme's activity may occur in part by phosphorylation and redox disruption of intermolecular disulphide bridges. The elevated levels of both EP24.15 activity and mRNA within testicular and neuroendocrine tissues indicate that EP24.15 gene expression is differentially regulated. In the present paper, we present a detailed analysis of the rat EP24.15 promoter region previously isolated and partially characterized in this laboratory. Employing site-directed mutagenesis to create a series of promoter deletions and full length promoter mutants, and measuring their activity in luciferase reporter gene and electrophoretic mobility-shift assays, we have shown that the transcription of the EP24.15 gene is differentially regulated in neuroendocrine and spermatid cell lines by transcription factor binding to SRY (sex-determining region Y), CAAT and CREB (cAMP-response-element-binding protein) promoter consensus sequences. The key to identifying the in vivo role of thimet oligopeptidase is likely to be found within the mechanisms by which it is regulated, and it is therefore of particular significance that EP24.15 expression is regulated by SRY and CREB/CREM (cAMP-response element modulator), the principle testes-determining protein and the major orchestrator of spermatogenesis respectively. PMID- 12911329 TI - A tobacco (Nicotiana tabaccum) calmodulin-binding protein kinase, NtCBK2, is regulated differentially by calmodulin isoforms. AB - A calcium (Ca2+)/calmodulin (CaM)-binding protein kinase (CBK) from tobacco (Nicotiana tabaccum ), NtCBK2, has been characterized molecularly and biochemically. NtCBK2 has all 11 conserved subdomains of the kinase-catalytic domain and a CaM-binding site as shown by other kinases, including Ca2+-dependent protein kinase and chimaeric Ca2+/CaM-dependent protein kinases. However, this kinase does not contain an EF-hand motif for Ca2+ binding, and its activity was not regulated by Ca2+. Whereas NtCBK2 phosphorylated both itself and other substrates, such as histone IIIS and syntide-2, in a Ca2+/CaM-independent manner, as also shown by OsCBK, a CaM-binding protein kinase from rice (Oryza sativa ), the kinase activity of NtCBK2 was greatly stimulated by Ca2+/CaM, whereas that of OsCBK was not. By molecular dissection analyses, the CaM-binding domain of NtCBK2 has been localized in a stretch of 30 amino acid residues at residue positions 431-460 as a 1-5-10 protein motif. Three tobacco CaM isoforms (NtCaM1, NtCaM3 and NtCaM13) used in the present study have been shown to bind to NtCBK2, but with different dissociation constants ( K(d)s), as follows: NtCaM1, 55.7 nM; NtCaM3, 25.4 nM; and NtCaM13, 19.8 nM, indicating that NtCBK2 has a higher affinity for NtCaM3 and NtCaM13 than for NtCaM1. The enzymic activity of NtCBK2 was also modulated differently by various CaM isoforms. Whereas the phosphorylation activity of NtCBK2 was shown by assay to be enhanced only approximately 2-3-fold by the presence of NtCaM1, the activity could be amplified up to 8-9-fold by NtCaM3 or 10-11-fold by NtCaM13, suggesting that NtCaM3 and NtCaM13 are better activators than NtCaM1 for NtCBK2. PMID- 12911330 TI - Fragmentation of extracellular matrix by hypochlorous acid. AB - The interaction of extracellular matrix with cells regulates their adhesion, migration and proliferation, and it is believed that damage to vascular matrix components is a factor in the development of atherosclerosis. Evidence has been provided for a role for the haem enzyme MPO (myeloperoxidase), released by activated monocytes (and possibly macrophages), in oxidative events within the artery wall. As MPO is released extracellularly, and is highly basic, it might be expected to associate with poly-anionic matrix components thereby localizing damage to these materials. In this study the reaction of the MPO-derived oxidant hypochlorous acid (HOCl) with extracellular matrix from vascular smooth muscle cells and healthy pig arteries has been examined. HOCl is rapidly consumed by such matrix samples, with the formation of matrix-derived chloramines or chloramides. The yield of these intermediates increases with HOCl dose. These materials undergo a time- and temperature-dependent decay, which parallels the release of sugar and protein components from the treated matrix, consistent with these species being important intermediates. Matrix damage is enhanced by species that increase chloramine/chloramide decomposition, with copper and iron ions being effective catalysts, and decreased by compounds which scavenge chloramines/chloramides, or species derived from them. The effect of such matrix modifications on cellular behaviour is poorly understood, though it is known that changes in matrix materials can have profound effects on cell adhesion, proliferation, growth and phenotype. The observed matrix modifications reported here may therefore modulate cellular behaviour in diseases such as atherosclerosis where MPO-derived oxidants are generated. PMID- 12911331 TI - Characterization of Amm VIII from Androctonus mauretanicus mauretanicus: a new scorpion toxin that discriminates between neuronal and skeletal sodium channels. AB - The venom of the scorpion Androctonus mauretanicus mauretanicus was screened by use of a specific serum directed against AaH II, the scorpion alpha-toxin of reference, with the aim of identifying new analogues. This led to the isolation of Amm VIII (7382.57 Da), which gave a highly positive response in ELISA, but was totally devoid of toxicity when injected subcutaneously into mice. In voltage clamp experiments with rat brain type II Na+ channel rNa(v)1.2 or rat skeletal muscle Na+ channel rNa(v)1.4, expressed in Xenopus oocytes, the EC50 values of the toxin-induced slowing of inactivation were: 29+/-5 and 416+/-14 nM respectively for AmmVIII and 2.6+/-0.3 nM and 2.2+/-0.2 nM, respectively, for AaH II interactions. Accordingly, Amm VIII clearly discriminates neuronal versus muscular Na+ channel. The Amm VIII cDNA was amplified from a venom gland cDNA library and its oligonucleotide sequence determined. It shows 87% sequence homology with AaH II, but carries an unusual extension at its C-terminal end, consisting of an additional Asp due to a point mutation in the cDNA penultimate codon. We hypothesized that this extra amino acid residue could induce steric hindrance and dramatically reduce recognition of the target by Amm VIII. We constructed a model of Amm VIII based on the X-ray structure of AaH II to clarify this point. Molecular modelling showed that this C-terminal extension does not lead to an overall conformational change in Amm VIII, but drastically modifies the charge repartition and, consequently, the electrostatic dipole moment of the molecule. At last, liquid-phase radioimmunassays with poly- and monoclonal anti (AaH II) antibodies showed the loss of conformational epitopes between AaH II and Amm VIII. PMID- 12911332 TI - Up-regulation of FLIP in cisplatin-selected HeLa cells causes cross-resistance to CD95/Fas death signalling. AB - Cisplatin-selected cervix carcinoma HeLa cell lines induced less apoptosis, and weaker activation by cisplatin or Fas-activating antibody, of mitochondrial associated caspase-9 and death receptor-mediated caspase-8 than did parental cells. Furthermore, less DISC (death-inducing signalling complex) was formed in cisplatin-selected cell lines than in parental cells. Ac-IETD-CHO (acetyl-Ile-Glu Thr-Asp-aldehyde), which has a certain preference for inhibiting caspase-8, or Fas-antagonistic antibody, significantly inhibited cisplatin-induced apoptosis in both parental and cisplatin-selected HeLa cell lines. These results imply that cell-surface death signalling is inducible by cisplatin; that reduction of this pathway is associated with drug resistance, and that cisplatin-selected cells acquire cross-resistance to cell-surface death signalling. Sequential up regulation of FLIP (FLICE-like inhibitory protein), but not Bcl-2, Bcl-x(L) or inhibitors of apoptosis protein (IAPs), was observed in resistant cells but not in parental cells. The inhibition of FLIP by FLIP antisense oligonucleotides promotes cisplatin and Fas-antibody-induced apoptosis. However, the modulation of apoptosis by FLIP antisense oligonucleotides in resistant cells is greater than that in parental cells. The presented data reveal that the up-regulation of FLIP may contribute to the suppression of apoptosis and thereby change cells that are resistant to cisplatin and Fas-mediated death signals. The results also show that cancer cells that have undergone long-term chemotherapy and become chemoresistant may change the FLIP level, becoming cross-resistant to death factors such as Fas. PMID- 12911333 TI - Allowed N-glycosylation sites on the Kv1.2 potassium channel S1-S2 linker: implications for linker secondary structure and the glycosylation effect on channel function. AB - N-glycosylation is a post-translational modification that plays a role in the trafficking and/or function of some membrane proteins. We have shown previously that N-glycosylation affected the function of some Kv1 voltage-gated potassium (K+) channels [Watanabe, Wang, Sutachan, Zhu, Recio-Pinto and Thornhill (2003) J. Physiol. (Cambridge, U.K.) 550, 51-66]. Kv1 channel S1-S2 linkers vary in length but their N-glycosylation sites are at similar relative positions from the S1 or S2 membrane domains. In the present study, by a scanning mutagenesis approach, we determined the allowed N-glycosylation sites on the Kv1.2 S1-S2 linker, which has 39 amino acids, by engineering N-glycosylation sites and assaying for glycosylation, using their sensitivity to glycosidases. The middle section of the linker (54% of linker) was glycosylated at every position, whereas both end sections (46% of linker) near the S1 or S2 membrane domains were not. These findings suggested that the middle section of the S1-S2 linker was accessible to the endoplasmic reticulum glycotransferase at every position and was in the extracellular aqueous phase, and presumably in a flexible conformation. We speculate that the S1-S2 linker is mostly a coiled-loop structure and that the strict relative position of native glycosylation sites on these linkers may be involved in the mechanism underlying the functional effects of glycosylation on some Kv1 K+ channels. The S3-S4 linker, with 16 amino acids and no N glycosylation site, was not glycosylated when an N-glycosylation site was added. However, an extended linker, with an added N-linked site, was glycosylated, which suggested that the native linker was not glycosylated due to its short length. Thus other ion channels or membrane proteins may also have a high glycosylation potential on a linker but yet have similarly positioned native N-glycosylation sites among isoforms. This may imply that the native position of the N glycosylation site may be important if the carbohydrate tree plays a role in the folding, stability, trafficking and/or function of the protein. PMID- 12911335 TI - Determination of gibberellins in fermentation broth produced by Fusarium verticilliodes MTCC 156 by high-performance liquid chromatography tandem mass spectrometry. AB - A method for the detection of gibberellins produced by Fusarium verticilliodes is described using HPLCMS/MS (HPLC tandem MS). A Hypersil (5 microm) octadecylsilane column with methanol/water as eluent in the ratio 3:1 at a flow rate of 0.5 ml/min was used. In the HPLCMS, GA(3) (gibberellic acid; m / z 346.3) eluted at retention time tr=3.08 min, with the corresponding mass chromatogram having peaks at m / z 346.7 and 328.8 corresponding to the M+ and M+-H2O ions respectively. The ethyl acetate extract from the broth, subjected to HPLCMS analysis under similar conditions, showed a constituent with tr=2.13 min, the mass chromatogram of which exhibited peaks at m / z 348.9 and 331.9 corresponding to the MH+ and MH+-H2O ions respectively. Comparison of the MS and MS/MS results (direct infusion) of an authentic sample of GA3 and the ethyl acetate extract from the broth revealed the formation of reduced GA3 in the broth. The present study, utilizing HPLCMS/MS, describes an improved methodology for the unambiguous determination and estimation of gibberellins from fermentation broth. PMID- 12911334 TI - A quantitative model of the generation of N(epsilon)-(carboxymethyl)lysine in the Maillard reaction between collagen and glucose. AB - The Maillard reaction between reducing sugars and amino groups of biomolecules generates complex structures known as AGEs (advanced glycation endproducts). These have been linked to protein modifications found during aging, diabetes and various amyloidoses. To investigate the contribution of alternative routes to the formation of AGEs, we developed a mathematical model that describes the generation of CML [ N(epsilon)-(carboxymethyl)lysine] in the Maillard reaction between glucose and collagen. Parameter values were obtained by fitting published data from kinetic experiments of Amadori compound decomposition and glycoxidation of collagen by glucose. These raw parameter values were subsequently fine-tuned with adjustment factors that were deduced from dynamic experiments taking into account the glucose and phosphate buffer concentrations. The fine-tuned model was used to assess the relative contributions of the reaction between glyoxal and lysine, the Namiki pathway, and Amadori compound degradation to the generation of CML. The model suggests that the glyoxal route dominates, except at low phosphate and high glucose concentrations. The contribution of Amadori oxidation is generally the least significant at low glucose concentrations. Simulations of the inhibition of CML generation by aminoguanidine show that this compound effectively blocks the glyoxal route at low glucose concentrations (5 mM). Model results are compared with literature estimates of the contributions to CML generation by the three pathways. The significance of the dominance of the glyoxal route is discussed in the context of possible natural defensive mechanisms and pharmacological interventions with the goal of inhibiting the Maillard reaction in vivo. PMID- 12911336 TI - The use of microcalorimetry to characterize tetanus neurotoxin, pertussis toxin and filamentous haemagglutinin. AB - Tetanus neurotoxin (TeNT), pertussis toxin (PT) and pertussis filamentous haemagglutinin (FHA) are major virulence factors of Clostridium tetani and Bordetella pertussis, which are the causative agents of tetanus and whooping cough respectively. Inactivated forms of these virulence factors are the protein components of vaccines against these diseases. Here we report microcalorimetric studies to characterize these proteins. The microcalorimetric titration curves of TeNT with micelles of gangliosides GD1b, GT1b and GQ1b were biphasic. For these gangliosides a high-affinity binding site (KD 45-277 nM) can be distinguished from a lower-affinity binding event (KD 666-1190 nM). This is direct evidence for multiple binding sites for gangliosides of the 1b series at TeNT as proposed by Emsley et al. [Emsley, Fotinou, Black, Fairweather, Charles, Watts, Hewitt and Isaacs (2000) J. Biol. Chem. 275, 8889-8894]. In agreement with previous reports, no binding was observed for gangliosides GM1, GM2, GM3 and GD2. The thermal denaturation of TeNT was characterized by two unfolding transitions centred around 57.4 and 62.4 degrees C. The conversion of TeNT into the toxoid form by formaldehyde treatment was accompanied by a large increase in Tm (the midpoint of protein unfolding transition, that is, the temperature at which half the protein is denatured and the other half is still present in its native form). Fetuin and asialofetuin bound to PT with similar affinities (KD 420 and 335 nM respectively). Binding was largely enthalpy-driven and counterbalanced by an unfavourable entropy change, indicating a loss of conformational flexibility. The latter could account for the observed inhibition of ATP binding after binding to fetuin. Furthermore, the molecular limits of mature PT subunit S5 were defined by MS and N-terminal peptide sequencing. The differential-scanning-calorimetry thermogram of FHA shows four well-resolved unfolding transitions, a finding consistent with the sequential denaturation of four structural domains. PMID- 12911338 TI - Public health: developing country perspectives. PMID- 12911337 TI - Expression and characterization of human interferon-beta1 in the methylotrophic yeast Pichia pastoris. AB - We describe the heterologous expression of a human interferon-beta1 in the methylotrophic yeast Pichia pastoris. Biologically active recombinant human interferon-beta1 (rHuIFN-beta1) was secreted from shake-flask-grown P. pastoris cells into the medium using the Saccharomyces cerevisiae alpha-mating factor prepro-leader sequence at the level of (1-3) x 10(5) i.u. (international units)/ml (6-12 mg/litre). An rHuIFN-beta1 with an N-terminal sequence identical with that of native HuIFN-beta1 was purified and the specific activity was determined (2-3 x 10(7) i.u./mg). It was found that the secreted recombinant protein was partially N-glycosylated. PMID- 12911339 TI - Decreased 2,3-diphosphoglycerate concentration in low cardiac output patients and its influence on the determination of in vivo p50. AB - We investigated whether 2,3-diphosphoglycerate (2,3-DPG) is altered in patients with low cardiac output and the influence of its concentration on the calculation of in vivo P(50). Biochemical and blood gas analysis were performed along with the measurement of cardiac output and body temperature in 13 patients submitted to cardiopulmonary bypass surgeries without the use of donor blood. In vivo P(50) was calculated using the measured (P(50m)) and the estimated 2,3-DPG (P(50e)). 2,3-DPG concentration was lower in these patients when compared to the values obtained in normal volunteers (6.9 +/- 2.2 vs. 11.9 +/- 2.4 microm/gHb). P(50m) was lower than P(50e) (21.6 +/- 1.1 vs. 30.1 +/- 1.2 mm Hg) at all time points. Our data show that in patients with low cardiac output, 2,3-DPG concentration is reduced. Therefore, in these patients, the use of standard values for this variable may introduce an error in the calculation of in vivo P(50). PMID- 12911340 TI - Is serum albumin a marker of nutritional status in hemodialysis patients without evidence of inflammation? AB - Hypoalbuminemia, a strong predictor of morbidity and mortality in hemodialysis patients, can be a consequence of a combination of malnutrition and inflammatory reactions. The purpose of this study was to analyze serum albumin as a marker of nutritional status in maintenance hemodialysis patients with no signs of inflammation. In a cross-sectional study, we selected 40 stable hemodialysis patients with normal levels of C-reactive protein (<0.8 mg/dL). The patients were classified as well nourished (65%) or malnourished (35%) according to the subjective global assessment. No significant differences were observed in serum albumin concentrations (immunoturbidimetric method) between well-nourished (4.3 +/- 0.3 g/dL) and malnourished (4.0 +/- 0.5 g/dL) patients, and the mean values were within the normal range in both groups. Albumin was inversely correlated with age (n=40; r=-0.32; P=0.02) and directly with energy intake (n=28; r=0.43; P=0.04). In this study, serum albumin did not discriminate well-nourished and malnourished hemodialysis patients without evidence of inflammation. PMID- 12911341 TI - Soluble Fas: a novel marker of inflammation in uremia. AB - BACKGROUND: Inflammation has been associated with atherosclerotic cardiovascular disease (CVD) and anemia in patients with end-stage renal disease (ESRD). Recent studies have shown that serum levels of soluble Fas (sFas), an antiapoptotic and proinflammatory molecule, are elevated in patients with cardiac disease and patients with ESRD. We therefore sought to investigate serum levels of sFas in uremic patients and its correlation with known markers of inflammation, anemia and CVD. METHODS: The study included 25 ESRD patients (14 on hemodialysis, 11 on CAPD), 27 patients with chronic kidney disease (CKD; creatinine clearance <50 ml/min/1.73 m2), and 14 normal control subjects. We measured serum levels of sFas, C-reactive protein (CRP), and albumin. We also investigated the association of serum sFas levels with the presence of CVD and with erythropoietin (EPO) dosage. RESULTS: Levels of sFas were elevated in CKD and ESRD patients compared to controls. sFas levels correlated negatively with creatinine clearance. In the dialysis patients, we observed that sFas levels were higher among those with CVD. Serum levels of sFas correlated with serum levels of CRP (r=0.31; P=0.03), serum levels of albumin (r=-0.35, P=0.02), and EPO dosage (r=0.51; P=0.009). CONCLUSION: These results suggest that sFas may be a marker of inflammation in CKD and ESRD patients. PMID- 12911342 TI - Glutaraldehyde-treated bovine pericardium: effects of lyophilization on cytotoxicity and residual aldehydes. AB - This work assesses the effect of lyophilization on the cytotoxicity and residual aldehyde concentration of glutaraldehyde-treated and lyophilized bovine pericardium (group A), comparing it to conventional glutaraldehyde-treated bovine pericardium (group B). Cytotoxicity was measured by incubating a pericardium sample from each group in saline and assessing the eluant's influence on cellular growth. Residual aldehydes were measured by HPLC. Although both groups' eluants exhibited some cytotoxicity, the eluant from group A was less cytotoxic, with a cytotoxicity index (IC50(%)) of 41%. Group B eluants all had marked cytotoxic effects; cell growth was 24.15% of the negative control at the most dilute eluant concentration (6.25%). The mean residual glutaraldehyde level was less in group A than in group B (2.36 +/- 0.11 and 9.90 +/- 3.70 g/l, respectively; n=3, P < 0.05), but residual formaldehyde levels did not differ. These results demonstrate that compared with conventional glutaraldehyde-treated bovine pericardium, lyophilized pericardium is less cytotoxic, with fewer glutaraldehyde residues. PMID- 12911343 TI - Noninvasive assessment of hemodynamic parameters in experimental stenosis of the ascending aorta. AB - We sought to noninvasively evaluate left ventricular (LV) function after cardiac hypertrophy induced by experimental stenosis of the ascending aorta. Male Wistar rats (70-90 g) underwent ascending aorta constriction by the surgical placement of a titanium clip (n=5) or sham operation (n=6). High-resolution bidimensional, pulsed-wave Doppler (PWD) and pulsed-wave tissue Doppler imaging (TDI) were performed 22 weeks after surgery. PWD was used to obtain mitral flow velocities, and TDI was used to obtain velocities along the septal mitral annulus and LV posterior wall. Clip placement produced myocardial hypertrophy with decreased systolic myocardial peak velocity in both the long and short axes. Increased myocardial mass, that is, posterior wall and septal thickness, was indicative of ventricular remodeling. Diastolic dysfunction was observed, with an increased early to late ratio of mitral velocities and increased left atrium dimension, consistent with a left ventricular restrictive filling pattern. PMID- 12911344 TI - A wound contraction experimental model for studying keloids and wound-healing modulators. AB - Preventing and treating hypertrophic and keloid scars is difficult because of the lack of knowledge about their genesis. Tissue repair can be studied with biocompatible matrices and ex vivo cultures of different cell types. We used an experimental model where collagen gels populated by human fibroblasts underwent progressive contraction, allowing the study of wound healing remodeling. The fibroblast-populated lattices showed the greater contraction of the gel populated by fibroblasts from keloids versus fibroblasts from normal skin. Moreover, fibroblast growth factor (FGF) and transforming growth factor beta (TGF-beta) involved in scar formation were added to the collagen gels populated by normal skin fibroblasts. TGF-beta caused an increase in gel contraction; FGF did not. The mean percentages of contraction of the gels populated by keloid fibroblasts were very similar to the percentages of gels populated by normal skin fibroblasts with added TGF-beta. These observations confirm the existing hypothesis that TGF beta may be involved in keloid formation. PMID- 12911345 TI - Commissural dehiscence and pannus formation of porcine heart valve bioprostheses. AB - From May 1976 to June 2001, 95 porcine heart valve bioprostheses (BPs) were explanted. Among them, 85 prostheses were available for an analysis of structural valve deterioration (SVD). There were 32 Hancock (H) BPs and 53 Carpentier Edwards (CE) BPs, with a mean implantation duration of 12 +/- 3.5 and 11.0 +/- 4 years, respectively. There were two extraordinary complications. One was commissural dehiscence (CD) of the CEBPs in 10/53 cases (18.9%), with a mean duration of 12.7 +/- 2.7 years. This was not observed in the HBPs. The other was pannus formation in both BPs. In the mitral position, it was 25/32 (78.1%) in HBPs and 37/53 (69.8%) in CEBPs. In the aortic position, it was 4/32 (1.3%) in the HBPs and 11/53 (20.8%) in the CEBPs. We conclude that CD could not be diagnosed preoperatively and anticoagulation is therefore indicated to prevent pannus formation of such prostheses, particularly in those located in the mitral position. PMID- 12911347 TI - A polyurethane vascular access graft and a hybrid polytetrafluoroethylene graft as an arteriovenous fistula for hemodialysis: comparison with an expanded polytetrafluoroethylene graft. AB - AIM: We evaluated a polyurethane vascular access graft (TVAG), a hybrid polytetrafluoroethylene graft (hPTFEG), and an expanded polytetrafluoroethylene graft (ePTFEG) for postoperative complications and graft patency in their use as prosthetic devices of vascular access for hemodialysis. METHODS: Between August 1993 and October 2001, we treated 200 patients in whom A-V fistulas were placed by the same surgeon. These were divided into the following four groups according to the type of blood access: 27 cases of ePTFEG, 23 cases of TVAG, 22 cases of hPTFEG, and 128 cases of an autogenous A-V fistula. We calculated the cumulative patency rates by the Kaplan-Meier method, including primary (problem-free) and secondary (revised or functional) patency rates. RESULTS: The hPTFEG group experienced few thromboses. The absence of perigraft edema in the TVAG group permitted the early use of the TVAG within a few postoperative days for hemodialysis. Among the three graft groups, the primary patency was the best in the hPTFEG group (94.7% at 1 year and 86.1% at 2 years), with a significant difference versus the ePTFEG group. In regard to secondary patency, hPTFEG had an excellent patency of 100% at 1 year and 90.9% at 2 years, and TVAG had a comparable patency with that of ePTFEG. CONCLUSION: The hPTFEG was considered superior to ePTFEG in terms of being complication-free and had the excellent 2 year secondary patency of 90.9%. TVAG, with a patency equal to that of ePTFEG, could be used immediately after implantation due to the absence of limb edema. PMID- 12911348 TI - Release of (1-->3)-beta-D-glucan from depth-type membrane filters and their in vitro effects on proinflammatory cytokine production. AB - To clarify the origin of (1-->3)-beta-D-glucan in blood products and assess the biological activity of filter extracts, we evaluated (1-->3)-beta-D-glucan extraction from depth filters used to process blood products and their in vitro effects on proinflammatory cytokine production from macrophages. Cellulose or nylon filters were analyzed for (1-->3)-beta-D-glucan using the Fungitec G test. To evaluate the biological activity of the filter extracts, Mono Mac 6 cells (a human macrophage cell line) were cultured with filter extracts with or without lipopolysaccharide, and tumor necrosis factor-alpha (TNF-alpha) and interleukin-1 beta (IL-1beta) concentrations in the culture media were measured. (1-->3)-beta-D Glucan was released from seven cellulose filters but the nylon filter level was undetectable. Proinflammatory cytokine production ranged from 74.3% to 119.0% of the control for TNF-alpha and 81.2% to 115.9% for IL-1beta. TNF-alpha and IL 1beta levels were low without lipopolysaccharide. The data indicate that (1-->3) beta-D-glucan in blood products is contaminated with the depth filters and that these filter extracts modulate proinflammatory cytokine production from macrophages. PMID- 12911346 TI - Mechanical properties of various z-stent designs: an endovascular stent-grafting perspective. AB - PURPOSE: To comparatively assess the mechani-cal behavior of various clinically relevant Z-stent designs. METHODS: A total of 16 Z-stents of original, biliary, spiral, and double-skirted designs (n=4 for each) were constructed using similar specifications for all. Stents were then evaluated for stiffness, snap opening force (SOF), flexibility, and displacement force using a novel tensiometer. Differences among the stents were determined using statistical methods. Stents explanted from dog aorta after a mean follow-up of 13 months were examined under a scanning electron microscope for surface defects. RESULTS: Forces required for about 50% reduction in diameter were 1.88 +/- 0.16 N, 3.81 +/- 0.21 N, 2.76 +/- 0.22 N, and 3.35 +/- 0.19 N for original, biliary, spiral, and skirted designs, respectively. Differences among the four designs were statistically significant at almost all points of measurement (P < 0.0001). Biliary and skirted designs showed higher SOF values in the early measurements. Stents explanted from dog aorta after a mean duration of 13 months showed no obvious corrosion or breakage in the wire struts. CONCLUSIONS: Significant differences exist among the various Z-stent designs in terms of their mechanical properties. Understanding them should help to select the appropriate stent for a given lesion. 316L stainless steel shows a favorable long-term tissue interaction. PMID- 12911349 TI - In vitro biocompatibility evaluation of the dynamic bubble trap. AB - The goal of this study was to evaluate the biocompatibility of the dynamic bubble trap (DBT) prior to the clinical trial. It was set up as an in vitro model, which simulates physiological conditions. Twenty runs were performed (ten with the DBT, ten without the DBT) at a blood flow of 3 l/min, each lasting 180 min. Fifteen blood parameters (hemogram, hemostasis, complement system, and cytokines) were measured at five time intervals. None of the tested parameters showed a statistically significant difference between the DBT and control group. The data assessed in this in vitro model show that the DBT has no adverse influence on hemocompatibility. It may be concluded that the DBT is a safe tool to be used in vivo. PMID- 12911350 TI - Measurement of the closing behavior of the bjork-shiley monoleaflet mechanical heart valve with an electrohydraulic total artificial heart. AB - When cavitation occurs near a material surface of a mechanical heart valve (MHV), pits on the surface of the MHV and hemolysis are caused. Therefore, it is very important to investigate the possibility of the occurrence of cavitation in an MHV. To study the possibility of cavitation occurrence in a 25 mm Bjork-Shiley monoleaflet, we analyzed the closing behavior of these valves. The closing event of these valves in the mitral and aortic positions was simulated in an electrohydraulic total artificial heart with a stroke volume of 100 ml. Tests were conducted under physiologic pressures at heart rates of 50, 60, 70, and 80 beats/min with cardiac outputs of 4.8, 5.9, 7.0, and 8.1 l/min, respectively. The disk-closing behavior was measured by a laser displacement sensor. The closing behaviors were investigated with various cardiac outputs and gravity direction. The maximum velocities of the aortic valve ranged from 0.8 to 0.9 m/s, and for the mitral valve ranged from 1.48 to 1.6 m/s. In aortic position valves, the maximum closing velocities were less than the reported cavitation thresholds, but the maximum closing velocities of the mitral valve were similar to the cavitation threshold. Therefore, we suggest that there should be the possibility of cavitation occurrence in the mitral valve of an electrohydraulic total artificial heart. PMID- 12911352 TI - Patient safety and quality: can anaesthetists play a greater role? PMID- 12911353 TI - Lateral tilt for pregnant women: why 15 degrees? PMID- 12911354 TI - Acute respiratory distress syndrome: an audit of incidence and outcome in Scottish intensive care units. AB - This prospective audit of incidence and outcome of the acute respiratory distress syndrome was conducted as part of the national audit of intensive care practice in Scotland. All patients with acute respiratory distress syndrome in 23 adult intensive care units were identified using the diagnostic criteria defined by the American-European Consensus Conference. Daily data collection was continued until death or intensive care unit discharge. Three hundred and sixty-nine patients were diagnosed with acute respiratory distress syndrome over the 8-month study period. The frequency of acute respiratory distress syndrome in the intensive care unit population was 8.1%; the incidence in the Scottish population was estimated at 16.0 cases.100,000(-1).year(-1). Intensive care unit mortality for acute respiratory distress syndrome was 53.1%, with a hospital mortality of 60.9%. In our national unselected population of critically ill patients, the overall outcome is comparable with published series (Acute Physiology and Chronic Health Evaluation II standardised mortality ratio = 0.99). However, mortality from acute respiratory distress syndrome in Scotland is substantially higher than in recent other series suggesting an improvement in outcome in this condition. PMID- 12911355 TI - Use of the bougie in simulated difficult intubation. 1. Comparison of the single use bougie with the fibrescope. AB - We studied the success rates for tracheal intubation in 64 healthy patients during simulated grade III laryngoscopy after induction of anaesthesia, using either the single-use bougie or oral flexible intubating fibrescope, both in conjunction with conventional Macintosh laryngoscopy. Patients were randomly allocated to either simulated grade IIIa or grade IIIb laryngoscopy, and also to one of the two study devices. Success rates for tracheal intubation (primary outcome measure) and times taken to achieve intubation (secondary outcome measure) were recorded. For the simulated grade IIIa laryngoscopy group, the fibreoptic scope was more successful than the bougie (16/16 successful intubations vs. 8/16; p = 0.02). For the simulated grade IIIb laryngoscopy group, the fibreoptic scope was also more successful than the bougie (8/16 successful intubations vs. 1/16; p = 0.02), but clearly use of the fibreoptic scope was not as successful as it had been in simulated grade IIIa laryngoscopy (p = 0.04). With either device, median (range) total tracheal intubation times for successful attempts with either grade of laryngoscopy were less than 60 s (19-109) and there were no clinically important differences. We conclude that the fibrescope used in conjunction with Macintosh laryngoscopy is a more reliable method of tracheal intubation than the single-use bougie in both types of grade III laryngoscopy. This finding has implications for the management of patients in whom grade III laryngoscopy is encountered unexpectedly after induction of anaesthesia, and also for the management of patients previously known to have grade III view at laryngoscopy. PMID- 12911356 TI - Use of the bougie in simulated difficult intubation. 2. Comparison of single-use bougie with multiple-use bougie. AB - We studied the success rates for tracheal intubation in 32 healthy, anaesthetised patients during simulated grade IIIa laryngoscopy, randomised to either the multiple-use or the single-use bougie. Success rates (primary end-point) and times taken (secondary end-point) to achieve tracheal intubation were recorded. The multiple-use bougie was more successful than the single-use one (15/16 successful intubations vs. 9/16; p = 0.03). With either device, median [range] total tracheal intubation times for successful attempts were < 54 [24-84] s and there were no clinically important differences between these times. We conclude that the multiple-use bougie is a more reliable aid to tracheal intubation than the single-use introducer in grade IIIa laryngoscopy. PMID- 12911357 TI - 1000 anaesthetic incidents: experience to date. AB - The anaesthetic incident reporting scheme in Leicester has been running for 11 years and 1000 incidents have now been reported. The scheme has successfully highlighted weaknesses where a procedural change has been able to prevent repetition. It has provided advance notification of problems which could be overcome by publicity and has been a source of educational cases. The experience of this scheme supports the use of a definition which does not include blame and allows the possibility of anonymous reporting. The scheme has evolved, driven by hospital decisions on reporting risk management cases, by inclusion of the Royal College of Anaesthetists' incident categories and by progressive refinements. Summary figures are given for the different categories of incident. These show marked similarities with previous studies. PMID- 12911358 TI - A prospective randomised controlled trial of capnography vs. bronchoscopy for Blue Rhino percutaneous tracheostomy. AB - A crucial step for successful percutaneous tracheostomy is the introduction of the needle and guide wire into the trachea. Capnography has recently been proposed as one way to confirm tracheal needle placement. In this randomised controlled study, we used capnography in 26 patients and bronchoscopy in 29 patients to confirm needle placement for percutaneous tracheostomy using Blue Rhino kit. The operating times and the incidence of peri-operative complications were similar for both groups. Capnography proved to be as effective as bronchoscopy in confirming correct needle placement. PMID- 12911360 TI - Validation of a pre-anaesthetic screening questionnaire. AB - We developed a screening questionnaire to be used by nurses to decide which patients should see an anaesthetist for further evaluation before the day of surgery. Our objective was to measure the accuracy of responses to the questionnaire. Agreement between questionnaire responses and the anaesthetist's assessment was assessed. For questions with a prevalence of 5 to 95%, the Kappa coefficient was used; percentage agreement was used for all other questions. Criterion validity was excellent/good for all questions with a prevalence between 5 and 95%, except for the question 'Do you have kidney disease?' For questions with prevalence < 5%, all demonstrated adequate criterion validity except the questions 'Has anyone in your family had a problem following an anaesthetic?' and 'If you have been put to sleep for an operation were there any anaesthetic problems?' Therefore, it is reasonable for nurses to use this questionnaire to determine which patients an anaesthetist should see before the day of surgery. PMID- 12911359 TI - A comparison of the forces exerted during laryngoscopy using disposable and non disposable laryngoscope blades. AB - The concern that re-usable equipment may be a source of cross-infection has prompted the increased use of disposable laryngoscope blades. We conducted a study investigating the duration of laryngoscopy and the peak force generated using various laryngoscope blades. Five blades were studied: one reusable Macintosh 3 blade, one disposable metal blade and three plastic disposable blades. Sixty anaesthetists performed laryngoscopy on a mannequin using each of the five blades presented in a random order. This was then repeated with a rigid collar applied to the mannequin. The mean force applied with the collar off with the metal blades was 32.8 and 30.8 N, and with the plastic blades 37.3, 39.6 and 41.5 N, respectively (p < 0.0001). The mean force applied with the collar on with the metal blades was 30.5 and 32.5 N, and with the plastic blades 35.5, 34.9 and 31.4 N, respectively (p < 0.0001). The mean duration of laryngoscopy with the collar off for the metal blades was 5.6 and 5.4 s, and with the plastic blades 10, 7.2 and 7.5 s (p < 0.0001). The duration of laryngoscopy with the collar applied for the metal blades was 7.1 and 7.5 s, and with the plastic blades 11.8, 9.7 and 9.0 s (p < 0.0001). The use of plastic blades results in both greater peak force and duration of laryngoscopy. PMID- 12911361 TI - Effects of hypertonic saline (7.5%) on extracellular fluid volumes in healthy volunteers. AB - This study evaluated the effects of 7.5% saline on plasma and other extracellular fluid volumes. After baseline measurements, eight healthy postmenopausal female volunteers received 4 ml.kg-1 of hypertonic saline over 30 min. After the fluid infusion, the volunteers were studied for 60 min. Plasma volume was measured using a dilution of 125-iodine-labelled human albumin. Extracellular water and cardiac output were measured by whole body impedence cardiography. The infused volume was 4 ml.kg-1 (average 260 ml). Plasma volume increased rapidly during the infusion (mean +/- standard deviation, 442 +/- 167 ml). At the end of the 1-h follow-up period, plasma volume had increased by on average 465 ml (SD 83). The increase of extracellular water at the end of infusion and at the end of study was 650 ml (SD 93) and 637 ml (SD 192), respectively. The highest serum sodium recorded in the volunteers was 158 mmol.l-1. The effect of 7.5% saline on plasma volume was rapid and lasted for at least 1 h. Plasma volume remained elevated by more than the infused volume at the end of the study. The increase in plasma and extracellular fluid volumes was partly achieved by mobilizing intracellular water to extracellular compartment. PMID- 12911362 TI - Out of our reach? Assessing the impact of introducing a critical care outreach service. AB - An outreach service was introduced in three surgical wards and the surgical high dependency unit in a large teaching hospital. A modified early warning score and callout algorithm were used to facilitate referrals to the team. Changes in unplanned admission rate to intensive care, length of stay, mortality rate and number of re-admissions following the introduction of outreach were sought. Following the introduction of the outreach service the emergency admission rate to intensive care fell from 58% to 43% (p = 0.05). These emergency patients had shorter lengths of stay (4.8 days vs. 7.4 days) and had a lower mortality (28.6% vs. 23.5%, p = 0.05). The re-admission rate also fell from 5.1% to 3.3% (p = 0.05). The outreach service had a significant impact on critical care utilisation. PMID- 12911363 TI - The effect of normobaric hyperoxia on cardiac index in healthy awake volunteers. AB - Fifteen healthy volunteers were exposed to a stepwise increase in FIO2 between 0.21 and 1.0, and their haemodynamic responses were measured with a non-invasive transthoracic bio-impedance monitor. There was mean reduction in cardiac index from 3.44 to 3.08 l.min-1.m-2 (10.7%, p < 0.001). The mean reduction in heart rate was from 77.3 to 69.1 beats.min-1 (10.5%, p < 0.001) and the mean systemic vascular index increased from 2062 to 2221 dyne.s-1.cm-5.m-2 (7.7%, p < 0.025). There were no significant changes in stroke index or mean arterial pressure. These changes are similar quantitatively and qualitatively to those previously reported by dye dilution techniques. PMID- 12911364 TI - Comparative evaluation of methods for ensuring the correct position of the tracheal tube in children undergoing open heart surgery. AB - The length of the trachea varies and is relatively short in children, it is therefore difficult to determine the correct depth of tracheal tube placement. In 85 children, the tube was placed using one of the following methods: (i) after deliberate endobronchial intubation, withdrawal to the carina was confirmed by auscultation, and the tube was then withdrawn a further 2 cm (auscultation group); (ii) as above, except that withdrawal to the carina was confirmed by a decrease in peak inspiratory pressure (pressure group); (iii) the tube was placed with a 3.0-cm mark at the vocal cords (mark group). The mean (SD) distance from the tip of the tube to the carina was 1.91 (0.81) cm in the auscultation group, and 1.93 (0.67) cm in the pressure group. These were not significantly different (p > 0.05) from targeted distance of 2 cm. In the mark group, the tube was located 2.30 (0.98) cm above the carina in children younger than 36 months and was further from the carina [6.16 (1.0) cm] in older children. In 20% of patients initially randomly allocated to the mark group, the mark could not be visualised. In conclusion, the methods described above effectively achieve adequate tracheal tube depth in children. PMID- 12911365 TI - Survey of abuse and violence by patients and relatives towards intensive care staff. AB - A postal survey of senior nurses in intensive care units in England and Wales was conducted. The aim was to ascertain the frequency of abusive and violent behaviour by patients and relatives towards intensive care staff, discover the perceived causes, effects and documentation of such behaviour and define the current and proposed security arrangements for intensive care units. Response rate was 94%. During the study period, verbal abuse of nurses by patients and by relatives occurred in 87% and 74% of intensive care units, respectively. The relevant figures for doctors were 65% and 59%, respectively. Nurses experienced physical abuse by patients and by relatives in at least 77% and 17% of intensive care units, respectively (doctors 38% and 8%). Illness was the main perceived cause of offences by patients whereas 'distress' (45%), alcohol (24%), sociopathic behaviour (27%) were the main putative causes amongst relatives. Whilst 43% of intensive care units have no security system at the door, staff awareness, training and communication skills may be the principle tools in reducing the frequency and consequences of violent and abusive behaviour. This survey probably underestimates the problem. PMID- 12911366 TI - The effects of concurrent atorvastatin therapy on the pharmacokinetics of intravenous midazolam. AB - Midazolam is a commonly used anaesthetic agent and is metabolised by the 3A4 isoform of the cytochrome P450 enzyme system. Atorvastatin is also metabolised by cytochrome P450 3A4 and, in vitro, atorvastatin inhibits the cytochrome P450 3A4 mediated metabolism of mexazolam. We hypothesised that concurrent administration of atorvastatin and midazolam would result in altered midazolam pharmacokinetics. Fourteen patients scheduled to undergo general anaesthesia for elective surgery were recruited in a matched pair design to receive intravenous midazolam (0.15 mg.kg-1). Of these patients, seven were taking long-term atorvastatin. Atorvastatin patients demonstrated a greater area under the curve (889.4 (standard deviation 388.6) ng-h.ml-1) vs. control patients (629.1 (standard deviation 197.2) ng-h.ml-1) (p < 0.05). Patients taking atorvastatin also demonstrated a decreased clearance (0.18 (standard deviation 0.08) l-kg. h-1) vs. control patients (0.27 (standard deviation 0.08) l-kg.h-1) (p < 0.05). This study suggests that chronically administered atorvastatin decreases the clearance of intravenously administered midazolam. PMID- 12911367 TI - The effect of variable-dose diazepam on dreaming and emergence phenomena in 400 cases of ketamine-fentanyl anaesthesia. AB - This randomised double-blind field study compared 400 anaesthetics using diazepam (0, 0.025, 0.5, 0.1, 0.175 mg.kg-1) with ketamine (1 mg.kg-1) and fentanyl (1 microg.kg-1) in Melanesian patients. Dreams were very common and generally positive in nature. A minimum of 0.1 mg.kg-1 of diazepam was needed to significantly reduce dreaming when compared with water (67.5% vs. 94.6%; p < 0.0001), and to significantly lower median (95% CI) emergence delirium scores (4 (3-4) vs. 6 (5-7)). Gender and age did not affect the rate of dreaming. Increasing the dose of diazepam did not improve the dream experience. Patient satisfaction scores were similar between groups. Increases in blood pressure and heart rate were greater in dreamers than in non-dreamers. All groups had high rate-pressure products but this was highest when diazepam was not used. Higher diazepam doses significantly reduced the increase in blood pressure and heart rate at 3 and 6 min postketamine. When used with ketamine and fentanyl, 0.1 mg.kg 1 of diazepam has favourable psychic and cardiovascular effects. Lower diazepam doses generally had little effect whereas larger doses did not enhance the benefits further. PMID- 12911368 TI - Syringe labelling. PMID- 12911369 TI - The canary is dead. PMID- 12911370 TI - Inducing anaesthesia in the operating theatre: staff and patient opinions. PMID- 12911371 TI - Subarachnoid haemorrhage following activated protein C for bacterial meningitis. PMID- 12911372 TI - Preventing damage to the tracheal tube during maxillary osteotomy. PMID- 12911373 TI - The intubating laryngeal mask and laser surgery. PMID- 12911375 TI - Don't pre-oxygenate with a 3-metre Bain circuit. PMID- 12911374 TI - Pulmonary aspiration during spontaneous ventilation through an Airway Management Device. PMID- 12911376 TI - Inadequate labelling of an unconventional device. PMID- 12911377 TI - Difficult airway alert. PMID- 12911378 TI - Nasotracheal intubation. PMID- 12911379 TI - Getting the tube in the oesophagus and oxygen in the trachea: preliminary results with the new supraglottic device (Cobra) in 28 anaesthetised patients. PMID- 12911380 TI - Sterilisation of laryngoscope blades. PMID- 12911381 TI - Sudden deaths during hip hemi-arthroplasty. PMID- 12911382 TI - Correct fixation of LMA ProSeal. PMID- 12911383 TI - Problems with the rapid powered injection of radiology contrast through multilumen catheters. PMID- 12911384 TI - Cornelia de Lange syndrome, hyperthermia and a difficult airway. PMID- 12911385 TI - General anaesthesia and the photosternutatory reflex. PMID- 12911386 TI - Unusual critical incident: chemical gas alert. PMID- 12911387 TI - Local anaesthetic antibacterial activity. PMID- 12911388 TI - Remifentanil sedation for parathyroidectomy. PMID- 12911389 TI - Dystonic reaction to cyclizine. PMID- 12911390 TI - The effect of auricular acupuncture on anaesthesia: a search for optimal design. PMID- 12911392 TI - Confirming the position of the thoracic epidural catheter--a valuable sign. PMID- 12911394 TI - Labour ward clocks. PMID- 12911393 TI - Aseptic precautions for inserting an epidural catheter. PMID- 12911395 TI - Thermal injury in pregnancy: anaesthetic considerations. PMID- 12911396 TI - Hypotension following combined spinal epidural anaesthesia. PMID- 12911397 TI - Anaesthetic management of the parturient with massive peripartum haemorrhage and fetal demise. PMID- 12911398 TI - A simple device as a guide to 15 degrees tilt during Caesarean section. PMID- 12911399 TI - Vegetarians and surgery. PMID- 12911400 TI - Drug mix-up during resuscitation. PMID- 12911401 TI - History of anaesthesia. PMID- 12911410 TI - Breastfeeding and allergic disease: a multidisciplinary review of the literature (1966-2001) on the mode of early feeding in infancy and its impact on later atopic manifestations. AB - BACKGROUND: Strategies to prevent children from developing allergy have been elaborated on the basis of state-of-the-art reviews of the scientific literature regarding pets and allergies, building dampness and health, and building ventilation and health. A similar multidisciplinary review of infant feeding mode in relation to allergy has not been published previously. Here, the objective is to review the scientific literature regarding the impact of early feeding (breast milk and/or cow's milk and/or formula) on development of atopic disease. The work was performed by a multidisciplinary group of Scandinavian researchers. METHODS: The search in the literature identified 4323 articles that contained at least one of the exposure and health effect terms. A total of 4191 articles were excluded mainly because they did not contain information on both exposure and health effects. Consequently, 132 studies have been scrutinized by this review group. RESULTS: Of the 132 studies selected, 56 were regarded as conclusive. Several factors contributed to the exclusions. The studies considered conclusive by the review group were categorized according to population and study design. CONCLUSIONS: The review group concluded that breastfeeding seems to protect from the development of atopic disease. The effect appears even stronger in children with atopic heredity. If breast milk is unavailable or insufficient, extensively hydrolysed formulas are preferable to unhydrolysed or partially hydrolysed formulas in terms of the risk of some atopic manifestations. PMID- 12911411 TI - T cell activation, from atopy to asthma: more a paradox than a paradigm. AB - During the last 15 years, it was largely shown that allergic inflammation was orchestrated by activated Th2 lymphocytes, leading to IgE production and eosinophil activation. Indeed, Th2 activation was shown to be necessary to induce allergic sensitization in animal models. In humans, a Th2 skewing was shown in atopic children soon after birth. In asthma, descriptive studies showed that Th2 cells were more numerous in patients than in controls. In addition, during specific allergen stimulation, an increase of Th2 cells was described in most cases. According to this Th2 paradigm, it was proposed that early avoidance of microbial exposure could explain the increase of atopic diseases seen in the last 20 years in developed countries, as the "hygiene hypothesis". Recently, it was proposed that early exposure to lipopolysaccharide (LPS) could be protective against atopic diseases. However, it is well established that exposure to LPS can induce asthma symptoms, both in animals and humans, although it induces a Th1 inflammatory response. In addition, most infections induce asthma exacerbations and Th1 responses. Recently, some studies have showed that some Th1 cells were present in asthmatic patients, which could be related to bronchial hyperreactivity. There is therefore an "infectious paradox" in asthma, which contributes to show that the Th2 paradigm is insufficient to explain the whole inflammatory reaction of this disease. We propose that the Th2paradigm is relevant to atopy and inception of asthma albeit a Th1 activation would account at least in part for bronchial hyperreactivity and asthma symptoms. PMID- 12911412 TI - Drug provocation testing in the diagnosis of drug hypersensitivity reactions: general considerations. PMID- 12911413 TI - Allergy: when does it begin and where will it end? PMID- 12911414 TI - Does occupational exposure to insects lead to species-specific sensitization? PMID- 12911415 TI - Exposure to high doses of birch pollen during pregnancy, and risk of sensitization and atopic disease in the child. AB - BACKGROUND: The role of maternal allergen exposure during pregnancy in sensitization and development of atopic disease in the child remains controversial. In the spring of 1993, extremely high levels of birch pollen were recorded in Stockholm, Sweden. In 1994, the corresponding pollen levels were low. The aim of this study was to assess the influence of exposure during pregnancy to high/low doses of birch pollen on the risk of sensitization and development of atopic disease in children. In addition, a comparison was made with children exposed to birch pollen in early infancy. METHODS: Three hundred and eighty-seven children with atopic heredity, born in Stockholm in July-October 1993 or 1994 (mothers exposed during pregnancy), were investigated at age 4.5 years. The children were clinically examined and were skin prick tested (SPT) with inhalant and food allergens. IgE antibodies (RAST) against birch pollen and recombinant birch pollen allergen (rBet v 1) were analysed in serum. A comparison was made with a similar group of children exposed during the same incident, but in the first 3 months of life, in 1993. RESULTS: The children of mothers high-dose exposed during pregnancy in 1993 tended to be more sensitized (SPT > or = 3 mm) to birch pollen than the children with low-dose exposure during the corresponding period in 1994 (7.6 and 4.6%, respectively, OR: 1.7; 95% CI: 0.7-4.1). A similar but weak tendency was seen for positive RAST analyses (> or =0.35 kU/l) against birch pollen and rBet v 1. Children of mothers high-dose exposed during pregnancy were significantly less sensitized to birch pollen than the children high-dose exposed in early infancy (17.9%, OR: 0.4; 95% CI: 0.2-0.7). There was an overall trend towards a slightly increased prevalence of bronchial asthma, allergic rhinoconjunctivitis and atopic dermatitis in the group with mothers high-dose exposed during pregnancy, compared to those with low exposure. CONCLUSION: Exposure of the mother during pregnancy to high levels of birch pollen resulted in a tendency towards increased risk of sensitization to the same allergen and symptoms of atopic disease in children with atopic heredity. Furthermore, our data indicate that exposure of the mother during pregnancy to inhalant allergens is less likely to result in sensitization in the child than exposure of the child in early infancy. PMID- 12911416 TI - Association of an early respiratory syncytial virus infection and atopic allergy. AB - BACKGROUND: Respiratory syncytial virus (RSV) causes postbronchiolitic wheezing but its role in allergic sensitization is controversial. The purpose of the study was to examine the effect of an early RSV infection on allergic sensitization. METHODS: Seventy-six subjects were examined 6-10 years after hospitalization for RSV infection during the first year of life. Fifty-one subjects (68%) attended clinical studies and 25 filled in a questionnaire. The study protocol included lung function, skin-prick and blood tests. The controls were matched for birth date and sex. RESULTS: Eight per cent of the subjects and 37% of the controls had at least one positive skin-prick test (SPT) (difference -35%, 95% CI -50 to -19%, P < 0.0001). Allergic rhinitis, atopic dermatitis and asthma occurred as often in both groups, but asthma had been diagnosed significantly earlier in the subjects than in the controls [mean age 3.0 years (SD 2.6) and 5.6 years (SD 3.0), difference 2.6 years, 95% CI 0.57-4.65, P = 0.014]. In a logistic regression analysis, RSV infection was associated with negative SPTs. CONCLUSIONS: An early RSV infection results in reduction of SPT positivity but not of occurrence of atopic diseases. This finding might explain why there is less atopic sensitization in countries with a greater probability of acquiring RSV infection at an early age. PMID- 12911417 TI - Characterization of airway nitric oxide in allergic rhinitis: the effect of intranasal administration of L-NAME. AB - BACKGROUND: Several studies have attempted to assess nasal nitric oxide (NO) levels in allergic rhinitis (AR). However, there seem to be differences in the results obtained. We therefore wanted to investigate this further by studying airway NO in AR and controls at several modalities, and also the effect of intranasal administration of the nitric oxide synthase (NOS) inhibitor NG-nitro-L arginine-methyl ester.HCl (L-NAME). METHODS: Airway NO was determined through repeated measurements at three flow rates of air (0.5, 3, and 9 l/min), using a single-breath method and a method of nasal aspiration, in 18 patients with birch pollen AR during season and in 18 controls. RESULTS: Patients with AR were characterized by no difference in nasal but higher orally exhaled NO and a larger interindividual spread in nasal and orally exhaled NO compared to controls. We also found a greater reduction in nasal NO after L-NAME in patients compared to controls. DISCUSSION: These results indicate that several factors determine the levels of nasal NO in rhinitis. NO production in the nasal mucosa of patients with AR may be upregulated. On the other hand, this increase could be counteracted by swelling of the mucosa and secretions resulting in impaired NO diffusion from, for example, the paranasal sinuses, where particularly high levels of NO have been found. Also, the high background levels of NO from constitutive sources in the nose may blunt smaller increases in mucosal NO output. CONCLUSION: It seems that the methods for measurement of nasal NO need to be improved and standardized before we can consider to use this test in monitoring inflammation in AR. PMID- 12911418 TI - Levocetirizine is effective for symptom relief including nasal congestion in adolescent and adult (PAR) sensitized to house dust mites. AB - BACKGROUND: Antihistamines are the most commonly prescribed class of medication for perennial allergic rhinitis (PAR). The primary objective of this study was to determine whether levocetirizine (Xyzal(R)), the active enantiomer of cetirizine, could achieve at least a 50% improvement in PAR symptoms compared to the placebo over the first week of treatment. METHODS: A total of 294 patients with PAR due to house dust mites were randomized in this 8-week double-blind, placebo controlled, multicentre trial to receive either levocetirizine 5 mg/day or placebo. Mean Total Four-Symptom Scores (T4SS) (nasal pruritus, ocular pruritus, rhinorrhoea and sneezing) were compared between treatment groups over weeks 1, 4 and 6. All individual symptom scores, including nasal congestion, were also studied. RESULTS: Levocetirizine showed an 86% improvement in T4SS over the first week of treatment and a 47% improvement over the entire treatment period compared with placebo. Absolute changes from baseline were 3.64 and 2.47 for levocetirizine and placebo, respectively. Individual symptom scores showed statistically significant (P < or = 0.01) differences in favour of levocetirizine for all study time-points. Nasal congestion was unexpectedly significantly improved (P < 0.001). The incidence of reported adverse events was comparable between treatment and placebo group. CONCLUSIONS: Levocetirizine 5 mg/day is an effective and well-tolerated treatment of PAR. In addition, levocetirizine is effective for the relief of nasal congestion. PMID- 12911419 TI - Effect of anti-nerve growth factor on early and late airway responses in allergic rats. AB - BACKGROUND: The increased production of nerve growth factor (NGF) has been associated with allergen-induced airway hyperresponsiveness and enhanced airway inflammation in experimental models of asthma. The aim of this study was to investigate whether a local application of anti-NGF to the lungs may affect the allergen-specific early (EAR) and late (LAR) airway responses to ovalbumin (Ova) of Ova-sensitized brown Norway rats. METHODS: Rats were sensitized systemically with Ova and were boosted twice intratracheally with Ova aerosol using a microsprayer. Two hours before every boost, the animals were pretreated either with aerosolized anti-NGF or with a control antibody. On day 21, all animals were challenged with inhalational Ova aerosol and pulmonary resistance was recorded in anesthetized, orotracheally intubated animals during the early and late asthmatic responses. In addition, differential cell counts from bronchoalveolar lavage and serum immunoglobulin E (IgE) levels were determined 48 h post-Ova challenge. RESULTS: Pretreatment with anti-NGF significantly attenuated the EAR but had no significant effect on the LAR. Serum IgE levels and inflammatory cell influx into the lungs were not affected by anti-NGF pretreatment. CONCLUSION: The data from this study suggest that NGF is directly involved in the development of the EAR without affecting the inflammatory airway response or LAR. PMID- 12911420 TI - Role and interpretation of total serum IgE measurements in the diagnosis of allergic airway disease in adults. AB - BACKGROUND: While total IgE measurements are often used in clinical practice, it is unclear how they should be interpreted for the diagnosis of allergic disorders. We studied whether total IgE may be used to rule out or predict sensitization and whether there are age or gender differences. METHODS: ROC curves were assessed in subjects with asthma or allergy symptoms from a general population sample. We studied predictive values and likelihood ratios. At least one positive skin test (Phazet) or specific IgE measurement (CAP) served as reference. RESULTS: High negative predictive values, suitable to rule out sensitization, were not found. In younger subjects, high total IgE levels strongly increase the probability of sensitization. The relationship between monosensitization and total IgE was less strong, but meaningful positive likelihood ratios were found at higher levels of total IgE. The discriminating ability of total IgE was better in the age group 20-44 than 45-70 years and comparable in males and females. CONCLUSION: Total IgE is not useful to rule out sensitization to common inhalant allergens. High total IgE may indicate a high probability of sensitization and may be useful to decide whether further investigation is warranted in patients with negative specific allergy tests to a panel of common inhalant allergens. PMID- 12911421 TI - Lack of human IgE cross-reactivity between mite allergens Blo t 1 and Der p 1. AB - BACKGROUND: The group 1 mite allergens are the most significant indoor allergens and they belong to the papain-like cysteine protease family. To date there is only one published report on the isolation and characterization of group 1 allergens from Blomia tropicalis mites. The aims of the study are to determine the cross-reactivity between group 1 allergens and to evaluate their clinical importance in allergic patients. METHODS: The full-length Blo t 1 gene was obtained by SMART RACE cDNA amplification method using gene-specific primers. The sequence alignment was performed using LOOK followed by three-dimensional homology modeling. The cDNA was expressed in Pichia pastoris as a secretory protein. Identification of native Blo t 1 in crude mite and spent mite medium extracts was done by Western immunoblot using monoclonal antibody. Allergenicity of recombinant Blo t 1 and native Der p 1 was examined by human IgE ELISA with 80 asthmatic sera. RESULTS: The cDNA sequence consists of 1105 base pairs, including 5'- and 3'-untranslating regions, encoding an open reading frame of 330 amino acid residues. The predicted molecular weight of the deduced protein was approximately 38 kDa. Blo t 1 shared 53 and 34% nucleotide and amino acid, respectively, sequence homology with Der p 1. Native Blo t 1 was detected in both crude mite and spent mite medium extracts, and its estimated molecular weight was about 26 kDa. The recombinant Blo t 1 reacted positively with IgE in 90 and 65% of sera from asthmatic children and adults, respectively, indicating that it is a major allergen. The correlation of human IgE reactivity between Blo t 1 and Der p 1 was low in these sera. CONCLUSION: The full-length cDNA encoding group 1 Blomia tropicalis mite allergen (designated as Blo t 1) has been characterized and expressed from local mites in Singapore. This fecal allergen showed high frequency of human IgE reactivity with asthmatic sera in the tropics and there was a low correlation of IgE reactivity between Blo t 1 and Der p 1. PMID- 12911422 TI - A further evaluation of the clinical use of specific IgE antibody testing in allergic diseases. AB - BACKGROUND: The evaluation and interpretation of the results from blood tests measuring specific immunoglobulin E (IgE) antibody concentration is currently made using the dichotomized result from the test despite a quantitative result is obtained. It has been shown that different levels of IgE antibodies, assessed by blood test and skin prick test, may have a relation to presence of symptoms, implying that there is more information in a quantitative result than in the dichotomous--positive or negative. OBJECTIVE: To investigate the clinical utility of quantification of IgE antibodies in the diagnosis of allergic patients and whether such procedure has any advantage to the presently dichotomously used sensitivity and specificity at a fixed cut-off. METHODS: Data from a previously published study (R. Paganelli, I.J. Ansoteugi, J. Sastre, C.-E. Lange, M.H.W.M. Roovers, H. de Groot, N.B. Lindholm, P.W. Ewan, Allergy, 1998; 53) analysing diagnosis of allergic patients in four different clinics were re-evaluated. In the original study consecutive patients with suspected IgE-mediated allergy had been examined and evaluated according to the clinical routine at each clinic, using case history, physical examination, skin tests and laboratory tests, except the test to be evaluated, and given a "doctors' allergen-specific diagnosis" as positive or negative. In the present study the relation between "doctors' allergen-specific diagnosis", expressed as pos/neg, and the quantitative levels of specific IgE antibody concentration was analysed using a logistic regression model. This presentation of results was also compared with the more common characteristics of sensitivity and specificity, and also with Receiver-operator characteristics (ROC) curves. RESULTS: The used logistic model described the relationship between allergen-specific diagnosis in each study and the levels of IgE antibodies. The shape of the curve illustrated the physicians' disposition for a positive diagnose in the study, in relation to the specific IgE antibody level. Differences in the shape of the curve was found both between allergens within clinics and between clinics for the same allergen. No association could be demonstrated between prevalence and shape of the curve. CONCLUSIONS: Conventional sensitivity/specificity figures or ROC concepts only use the qualitative statement of whether IgE is present or not. A risk assessment using the quantitative level of IgE antibody to an allergen increases the utility of the information in clinical context compared with a qualitative statement of whether IgE is present or not. The quantification demonstrated the link between specific IgE antibodies and allergic reactions. The use of objective, well performing quantitative tests should help improve diagnostic accuracy and might provide a way for the patient to understand and manage his or her daily situation and risk for reactions. PMID- 12911423 TI - Detection of specific IgE antibodies in the sera of patients allergic to birch pollen using recombinant allergens Bet v 1, Bet v 2, Bet v 4: evaluation of different IgE reactivity profiles. AB - BACKGROUND: Birch pollen is a significant cause of immediate hypersensitivity among susceptible subjects in temperate climates, affecting 5-54% of the population in western Europe. We examined the specific serum IgE antibodies towards recombinant allergens Bet v 1, Bet v 2 and Bet v 4 in birch-sensitive patients from the province of Cuneo, north-west Italy. METHODS: Sera were obtained from 372 patients with symptomatic birch pollen-induced allergic rhinitis and/or asthma. A subgroup of these patients suffered from oral allergy syndrome after eating apple. Their sera were evaluated for specific IgE against natural birch pollen and apple extract, as well as Bet v 1, Bet v 2 and Bet v 4 using Pharmacia CAP system (Pharmacia, Uppsala, Sweden). RESULTS: Of 372 patients 215 (57.80%) had serum-specific IgE towards Bet v 1. A total of 166 sera (44.62%) contained serum-specific IgE to Bet v 2, while Bet v 4 IgE reactivity was documented in 35 subjects (9.41%). Moreover, 146 (39.25%) patients were monosensitized to Bet v 1; 96 (25.81%) patients were monosensitized to Bet v 2; only four sera (1.08%) contained specific IgE towards Bet v 4. Thirty-nine sera (11.02%) did not contain specific IgE to these individual birch pollen allergens. Of course, all 372 sera (100%) had specific IgE against natural birch pollen extract, of which 162 (43.55%) contained specific IgE to apple extract (75.35% of Bet v 1 positive sera). CONCLUSION: In this study we observed that three birch pollen recombinant allergens alone, could sufficiently identify 90% of birch pollen-sensitive patients. Therefore, for a more precise IgE profile of patients allergic to birch, further purified birch pollen allergens (i.e. Bet v 6, Bet v 7, Bet v 8) will be required. PMID- 12911424 TI - 15-year aeroallergen records. Their usefulness in Athens Olympics, 2004. AB - INTRODUCTION: About 5-25% of 16 000 athletes involved in preparation for the Athens 2004 Olympics may encounter respiratory allergy (asthma and rhinoconjunctivitis) triggered by exposure to aeroallergens (pollen and fungi spores). AIM: Provide information about circulating aeroallergens in three Olympic cities and ensure safety for the allergic athletes who will visit Greece from January to September 2004. METHODS: Aeroallergens were recorded using a Burkard volumetric trap. The most frequently implicated pollen (cypress, hazel, wall pellitory, plane, olive, grasses, goosefoot and mugwort) and fungi spores (Alternaria spp., Cladosporium spp.) are presented. These data derive from a 15 year database created by the Pulmonary Department, Thessaloniki, a 2-year database (University of Heraklion-Crete) and a 6-year database by the Pediatric Department, Athens. The above data and the current aeroallergen counts will be continuously announced by the mass media and Internet during the Athens 2004 Olympics. RESULTS: Are expressed as: (A) Presentation (Graphs, Tables) of the fluctuation of mean daily values of pollen grains or fungi spores/m3. (B) Presentation (Tables) of the start, peak and end time of aeroallergen circulation. Peak pollen concentrations were observed between March and May (athletes preparation time). During the 2004 Olympics (August-September) relatively high concentrations of goosefoot, mugwort, Alternaria and Cladosporium will be observed. Aeroallergens circulate in Athens 10-15 days earlier than in Thessaloniki and 10 days later than in Heraklion. CONCLUSIONS: The data presented in the study are expected to help allergic Olympic athletes achieve peak performance under prophylactic measures. PMID- 12911425 TI - Pet-keeping in early childhood and airway, nose and skin symptoms later in life. AB - BACKGROUND: It is discussed whether exposure to pets during childhood is a risk or a protective factor for sensitization and allergic symptoms. The aim of this study was to investigate the association between pet-keeping at time of birth and allergic symptoms in airways, nose and skin among young children in Sweden. METHOD: A questionnaire was sent to the parents of 14 077 children (1-6 years), the focus being on allergic symptoms, home environment and other background factors including pet-keeping and avoidance behaviour. The response rate was 79%. RESULTS: Almost one-tenth of the population had got rid of pets because of allergy in the family, and 27.3% reported "avoidance" behaviour towards pets. In a cross-sectional analysis current pet-keeping was "protective", but this may be due to the fact that people avoid exposing their child to something that they believe is a risk factor for allergies. Pet-keeping at the time of birth was associated with "wheezing", "asthma" and "rhinitis on pet-exposure" later in life for children from families with an "avoidance" behaviour, and was not "protective" for other children. There was also an indication of a dose-response relationship between the number of types of furred pets at time of birth and later symptoms in analyses adjusted for avoidance behaviour or current pet keeping. CONCLUSION: The distribution of pet-keeping in the population is largely explained by avoidance behaviour, meaning that those who have pets mainly are those who can stand them, indicating a "healthy pet-keeping effect". PMID- 12911426 TI - Relationship between cutaneous allergen response and airway allergen-induced eosinophilia. AB - BACKGROUND: Determinants of changes in airway caliber after allergen challenge include nonallergic airway responsiveness, immune response and dose of allergen given. However, determinants of the airway inflammatory response to allergens remain to be determined. AIM: To assess the relationship between skin reactivity to airborne allergens and lower airway eosinophilic response to allergen exposure in asthma and allergic rhinitis. METHODS: Forty-two subjects with mild allergic asthma (mean age 24 years) and 14 nonasthmatic subjects with allergic rhinitis (mean age 25 years) had allergen skin prick tests and titration with the allergen chosen for subsequent challenge. On a second visit, 31 asthmatic subjects had a conventional challenge while 11 asthmatic subjects and all rhinitic subjects had a low-dose allergen challenge over four subsequent days. Induced sputum samples were obtained at 6 and 24 h after the conventional challenge and at days 2 and 4 of the low-dose challenge. RESULTS: In the asthmatic group, there was a weak correlation between wheal diameter induced by the concentration used for challenge and increase in eosinophils 6 h postconventional challenge (r = 0.372, P = 0.05), but no correlation was observed following the low-dose challenge. Rhinitic subjects showed a correlation between wheal diameter with the allergen dose used for bronchoprovocation and increase in eosinophils at day 2 of low dose (r = 0.608, P = 0.02). CONCLUSION: This study suggests that immediate immune responsiveness to allergen, assessed by the magnitude of the skin response, is a significant determinant of allergen-induced airway eosinophilia and can help to predict the airway inflammatory response. PMID- 12911429 TI - Anaphylaxis in a child playing in tall grass. PMID- 12911427 TI - Diagnostic value of scratch-chamber test, skin prick test, histamine release and specific IgE in birch-allergic patients with oral allergy syndrome to apple. AB - BACKGROUND: The aim of the study was to examine the diagnostic value of skin prick test (SPT), scratch-chamber test (SCT), histamine release (HR) and specific immunoglobulin E (IgE) in birch-allergic patients with oral allergy syndrome to apple. METHODS: Ten birch-allergic patients with oral allergy syndrome to apple and 10 control subjects were included. All were tested with SPT, SCT, HR and specific IgE [CAP, Pharmacia, Sweden and Magic Lite (ML), ALK-ABELLO, Denmark]. RESULTS: The SPT with apple, acetone extract of apple (A72) and commercial apple extract showed sensitivities of 0.80, 0.90 and 0.10, respectively. The SCT with the same extracts showed sensitivities of 0.30, 0.50 and 0.20, respectively. The sensitivity of specific IgE to apple were 0.90 (CAP) and 0.10 (ML). The sensitivity of the HR test was 90% (A72), and 25% using the commercial extract. CONCLUSION: The SPT and HR test with apple and A72 showed a good diagnostic value with a sensitivity of more than 70% and a specificity of 100%. The SCT showed a poor sensitivity to apple, A72 and commercial apple extract. The ML test was not suitable in detecting specific IgE to apple compared with the CAP test. In daily practice a detailed case history about symptoms of oral allergy syndrome combined with a SPT with fresh apple peel or A72 will be useful. PMID- 12911430 TI - Oral wheat flour anaphylaxis related to wheat alpha-amylase inhibitor subunits CM3 and CM16. PMID- 12911431 TI - Occupational contact allergy to ceftiofur. PMID- 12911432 TI - Occupational asthma and allergic rhinitis due to xerographic toner. PMID- 12911433 TI - Nickel sulphate, food additives and seronegative arthritis: is there any relationship? PMID- 12911434 TI - Serial blood and urine levels of EDN and ECP in eosinophilic colitis. PMID- 12911436 TI - The development of perinatal audit: 20 years' experience. PMID- 12911437 TI - Homocysteine-induced enhancement of spontaneous contractions of myometrium isolated from pregnant women. AB - BACKGROUND: Although many associations have been demonstrated between hyperhomocysteinemia and pregnancy complications, such as spontaneous abortion, preterm labor, preeclampsia and low birthweight, it is still not clear whether hyperhomocysteinemia is the cause or the consequence of these pregnancy complications. The aim of this study was to investigate the effects of homocysteine on the spontaneous contractility of isolated pregnant human myometrium. METHODS: Myometrium samples obtained from women undergoing elective caesarean section were suspended in a jacketed organ bath containing Krebs' solution at 37 degrees C (pH 7.4), continuously gassed with 95% O2 and 5% CO2. After manifestation of spontaneous contractions under 2 g of resting tension, homocysteine was applied to the organ bath, and amplitude and frequency of contractions were evaluated at 20-min intervals. Statistical analysis of amplitude and frequency of the contractions was performed using the Kruskal Wallis analysis of variance test. RESULTS: Application of 0.1 mm homocysteine had no significant effect on either frequency (4.63 +/- 0.42 vs. 4.01 +/- 0.53 for the control period; p > 0.05) or amplitude (3.20 +/- 0.07 g vs. 3.15 +/- 0.07 g; p > 0.05) of spontaneous contractions, while 1 and 2 mm homocysteine significantly increased the frequency (8.54 +/- 0.62 vs. 3.9 +/- 0.44, p < 0.02, and 12.32 +/- 0.72 vs. 3.96 +/- 0.51, p < 0.001, respectively) of spontaneous contractions but caused only a modest change in amplitude (2.92 +/- 0.04 g vs. 2.87 +/- 0.05 g, p > 0.05, and 3.02 +/- 0.06 g vs. 2.93 +/- 0.05 g, p > 0.05, respectively). CONCLUSION: Results from this study indicate for the first time that homocysteine causes enhancement of spontaneous contractions of myometrium derived from pregnant women. PMID- 12911438 TI - Effect of antenatal corticosteroid administration on Doppler flow velocity parameters in pregnancies with absent or reverse end-diastolic flow in the umbilical artery. AB - OBJECTIVE: Evaluation of alterations of Doppler flow velocity waveforms after antenatal corticosteroid therapy. METHODS: Pulsatility index of the umbilical artery, middle cerebral artery and ductus venosus waveforms was measured in high risk pregnancies complicated by umbilical AREDV directly prior to and within 24 or 48 hours after application of the first dose of a steroid treatment course. RESULTS: Return of a positive end-diastolic flow after umbilical AREDV or conversion of REDV into zero-flow was detected in 9 of 20 (45%) pregnancies after steroid application. Neither in the group which showed altered umbilical end diastolic flow nor in the total of patients investigated was there any significant change in the PI of the umbilical artery or the ductus venosus waveforms. In contrast, there was an overall drop of pulsatility in the middle cerebral artery. CONCLUSIONS: Antenatal corticosteroid administration could increase cerebral perfusion but has no influence on umbilical and venous blood flow. PMID- 12911439 TI - Maternal plasma levels of cytokines in normal and preeclamptic pregnancies and their relationship with diastolic blood pressure and fibronectin levels. AB - BACKGROUND: To determine the plasma concentrations of placental growth factor (PLGF), vascular endothelial growth factor (VEGF), transforming growth factor beta1 (TGF-beta1), soluble tumor necrosis factor alpha receptor (sTNFp55), interleukin-2 receptor (IL-2R), and interleukins 6 and 10 (IL-6, IL-10) in normotensive and preeclamptic women, and to evaluate the correlations between these cytokines and the diastolic blood pressure and fibronectin levels. METHODS: A prospective case-control study. Thirty-five women with preeclampsia were compared with 34 healthy women with uncomplicated pregnancies. Peripheral venous blood samples were obtained and plasma levels of PLGF, VEGF, TGF-beta1, sTNFp55, IL-2R, IL-6 and IL-10 were measured by an enzyme-linked immunoassay and fibronectin by a radial immundiffusion technic. RESULTS: In preeclampsia PLGF and VEGF levels were significantly lower, and TGF-beta1, sTNFp55, IL-2R, IL-6 and IL 10 levels were significantly higher than in normotensive pregnancy (p < 0.001). The plasma levels of PLGF and VEGF significantly decreased, whereas TGF-beta1, sTNFp55, IL-2R, IL-6 and IL-10 levels significantly increased with the increments in diastolic blood pressure and fibronectin levels (p < 0.001). CONCLUSIONS: Altered concentrations of various cytokines might explain the shallow placentation and endothelial cell dysfunction described in preeclampsia. The clinical severity of preeclampsia seems to correlate with the severity of the cytokine abnormalities. PMID- 12911440 TI - Experiences with fetomaternal alloimmune thrombocytopenia at a Swedish hospital over a 10-year period. AB - BACKGROUND: This is a descriptive study of the management and outcome of 18 cases of fetomaternal alloimmune thrombocytopenia (FMAIT) treated from 1991 to 2001. MATERIAL AND METHODS: Management of the disease changed over the years from cordocentesis in the 20-24th week of gestation, platelet transfusions and immunoglobulin to a less invasive management consisting of only blind administration of immunoglobulin and predelivery cordocentesis. RESULTS: Three of the fetuses were treated with intrauterine platelet transfusions. Two of these were delivered by emergency cesarean section due to failed transfusions and the third fetus died as a result of the procedure. Nine mothers were treated with immunoglobulin intravenously. Four of these delivered thrombocytopenic children. Three women did not want to undergo any treatment, and all newborns had low platelet counts. Two fetuses died, one in conjunction with a platelet transfusion and the other in utero before treatment was commenced. All the other children did well despite the fact that some of them were severely thrombocytopenic at birth. CONCLUSIONS: Due to the limited number of patients, the present material does not allow any far reaching conclusions. Our experience is that a non-invasive management can be practiced in cases of FMAIT. The value of performing cordocentesis and platelet transfusions in the second trimester is doubtful in view of the risk for the fetus and the limited amount of information it provides for management of the individual case. PMID- 12911441 TI - New score indicating placental vascular resistance. AB - BACKGROUND: Umbilical artery Doppler velocimetry is a routine method for fetal surveillance in high-risk pregnancy. Uterine artery Doppler seems to give comparable information, but it can be difficult to interpret as there are two arteries, which might show notching and/or increased pulsatility index (PI) as signs of increased vascular impedance. Combining the information on vascular resistance on both sides in a new score might simplify and improve evaluation of placental circulation. METHODS: Uterine and umbilical artery Doppler velocimetry was evaluated in 633 high-risk pregnancies. The managing clinician was informed only about the umbilical artery flow. The umbilical artery flow spectrum was semiquantitatively divided into four blood flow classes (BFC), expressing signs of increasing vascular resistance. The uterine artery Doppler flow spectrum was divided into five uterine artery scores (UAS), taking into account presence/absence of notching and/or increase in PI. By adding UAS to BFC, a new placental score (PLS) was constructed with values ranging from 0 to 7, indicating general placental vascular resistance. The scores were related to three outcome variables: small-for-gestational age (SGA), premature delivery (<37 weeks), and cesarean section. RESULTS: All three score systems showed a significant relationship between signs of increasing vascular resistance and outcome. The new PLS showed the best association to adverse outcomes, with optimal cut-off at values exceeding score 3. CONCLUSION: Doppler velocimetry on both sides of the placenta showed a strong relationship to an adverse outcome of pregnancy. The new PLS showed a better relationship to adverse perinatal outcome than the BFC and the UAS. The PLS can simplify evaluation of uteroplacental and fetoplacental Doppler velocimetry. PMID- 12911442 TI - Multimodal intervention raises smoking cessation rate during pregnancy. AB - BACKGROUND: The aim was to study the effect of a multimodal smoking cessation intervention regimen on a number of pregnant smokers. METHODS: A prospective intervention study was designed where participants were allocated to intervention or control based on their birth date. The study included 647 pregnant smokers. The intervention group (n = 327) received initial individual smoking cessation counseling supplemented by an invitation to join, individually or in a group, a smoking cessation program with nicotine replacement therapy as a voluntary option. Intervention was designed as an integral part of the midwives' prenatal care. All pregnant smokers in the usual care group (n = 320) received standard counseling from a midwife. Outcome was self-reported smoking cessation in the 37th week of pregnancy and the reported cessation was validated by cotinine saliva concentration. RESULTS: Self-reported cessation rates during pregnancy were significantly higher in the intervention group (14%) than in the group receiving usual care (5.0%) (p < 0.0001) (Fisher's exact test). Cotinine validated cessation rates during pregnancy were significantly higher among the former (7%) than the latter (2%) (p = 0.003). The adjusted odds ratio (OR) for smoking cessation was 4.20 (95% CI 2.13-8.03). Logistic regression analysis showed a significant positive association of smoking cessation with low caffeine consumption in pregnancy, many years in school, no exposure to passive smoking outside the home, and previous attempts to stop smoking. CONCLUSIONS: A multimodal intervention regimen with initial individual counseling supplemented by an invitation to join a smoking cessation program with nicotine replacement therapy as a voluntary option markedly increased cessation rates during pregnancy. PMID- 12911443 TI - Relationship between bone mineral density changes in pregnancy and maternal and pregnancy characteristics: a longitudinal study. AB - OBJECTIVE: The study aims to verify whether a progressive fall in bone mineral density (BMD) values can be demonstrated using quantitative ultrasound measurements of the os calcis. The BMD change during the pregnancy was then correlated with other maternal and pregnancy characteristics to identify any high risk factors for bone loss in pregnancy. METHODS: Consecutive patients were recruited from a low-risk obstetric clinic over a period of 9 months. BMD measurements were performed at the os calcis before 18 weeks, between 28 and 32 weeks, and at 36-38 weeks using a Hologic Sahara Clinical Bone Sonometer system. A computer-derived BMD value was obtained with each measurement. Body fat composition was also measured using a Tanita 501 bioimpedance assay system. RESULTS: In a cohort of 780 patients, a mean fall in BMD of 0.040 g/cm2 was demonstrable across the trimesters. The difference in BMD across the three serial measurements was highly significant (p < 0.001). Univariate analysis showed that those with a low initial BMD, glucose intolerance and high body fat accumulation weight gain during the pregnancy had lower BMD loss, while gestational hypertension and obesity had no impact on the degree of BMD loss. Entering these parameters in a logistic regression analysis showed that the impact of glucose intolerance was lost, but that the other factors remained significant. A linear regression model of quantitative variables showed that only fat accumulation (p = 0.03) and early pregnancy BMD values (p < 0.001) remained significant factors associated with BMD loss. CONCLUSION: A gradual fall in BMD was demonstrable using ultrasound measurement of the os calcis from early to late pregnancy. Of the various parameters identified as significant factors affecting BMD loss in pregnancy, a low initial BMD in early pregnancy and high body fat accumulation during pregnancy appeared to be related to lower BMD loss. PMID- 12911444 TI - Prospect for vaginal delivery of growth restricted fetuses with abnormal umbilical artery blood flow. AB - BACKGROUND: The best mode of delivery in cases of intrauterine growth restriction (IUGR) with umbilical artery blood flow changes is not well elucidated. OBJECTIVE: To evaluate outcome in IUGR with umbilical artery blood flow changes planned for vaginal delivery after a negative oxytocin challenge test (OCT). METHODS: In 84 term singleton pregnancies with suspected IUGR and no unanimous indication for abdominal delivery, Doppler velocimetry and OCT were performed. Positive OCT cases were delivered by cesarean section, negative OCT cases planned for vaginal delivery. RESULTS: Umbilical artery Doppler velocimetry was normal in 51 cases (normal group) and abnormal in 33 cases (increased pulsatility index with maintained forward diastolic flow). Gestational age at delivery was shorter (p = 0.008), positive OCT more common (33% vs. 16%; p = 0.06), and vaginal delivery less common (40% vs. 63%; p = 0.04) in the abnormal blood flow group compared with the normal flow group. When in labor, 68% in the abnormal flow group and 76% in the normal flow group delivered vaginally (p = 0.6). One baby had a lethal malformation and another suffered meconium aspiration and pneumothorax, but was discharged home healthy. CONCLUSIONS: The vaginal delivery rate was significantly lower in the abnormal flow group compared with the normal flow group, but in cases finally destined for a trial of labor the vaginal delivery rates were similar. There was no indication that any fetus was exposed to detrimental hypoxia or distress. PMID- 12911445 TI - Elective cesarean section vs. spontaneous delivery: a comparative study of birth experience. AB - BACKGROUND: To investigate birth experience and medical outcome in women with elective cesarean section (CS) compared with women with intended vaginal delivery. METHODS: A total of 1050 pregnant women were included in this prospective trial. Psychological factors, pain levels and birth experience were investigated using a self-designed questionnaire and three established psychological tests in gestational week 38, and 3 days and 4 months postpartum. In addition, medical data were evaluated from the records. RESULTS: Out of 903 women with planned vaginal birth, in 484 women (53.6%) minimal perineal surgery had to be performed after birth, 41 women (4.5%) had vacuum deliveries, and in 93 cases (10.3%) emergency CS had to be performed. In the 147 elective CS (103 based on medical and 44 on psychological factors), a significantly lower rate of maternal and fetal complications was observed when compared with vaginal birth (5.4% vs. 19.3%; p < 0.0001). Birth experience (Salmon test) was significantly better in elective CS compared with vaginal delivery, but worse in women with emergency CS and worst in those with vacuum delivery. We found that 83.5% of women with vaginal delivery would choose the same mode of birth again, 74.3% of women with CS on demand, and 66% of women with medically necessary CS. Only 30.1% of women with emergency CS wanted to receive CS at the next birth. CONCLUSIONS: Elective CS is a safe and psychologically well tolerated procedure. The results are comparable with uncomplicated vaginal delivery and far superior to secondary intervention such as vacuum delivery or emergency CS. PMID- 12911446 TI - Undiagnosed term breech: impact on mode of delivery and neonatal outcome. AB - BACKGROUND: The purpose of this study was to compare delivery mode and neonatal outcome in breech pregnancies diagnosed antepartum (Dx group) with those diagnosed on admission for delivery or intrapartum (No-Dx group). METHODS: During an 11-year period, breech pregnancies >/=34 weeks' gestation with a live nonanomalous fetus were reviewed. The Dx group was matched for gestational age, parity, and maternal age to the No-Dx group (n = 256) and compared using the chi2 test, the Mc-Nemar test, and the Wilcoxon rank test. A p-value of < 0.05 was considered significant. RESULTS: Breech type and birthweights were similar in the two groups. In the No-Dx group, 19.5% did not receive prenatal care. More patients in the No-Dx group were admitted with cervical dilation > 4 cm (39.4% vs. 27.0%, p = 0.004), whereas 17.1% of patients in the Dx group were admitted for elective cesarean delivery at term. There was a higher cesarean delivery rate in the Dx group (64.1% vs. 50.8%, p = 0.003), specifically for arrest disorders (15.2% vs. 6.9%, p = 0.008). The neonatal outcome in the two groups was similar regarding Apgar scores, intensive care nursery admission, need for mechanical ventilation, neonatal death, and length of nursery stay. CONCLUSIONS: Antepartum diagnosis of breech presentation decreases the threshold for cesarean delivery. Failure to diagnose breech antepartum does not compromise neonatal outcome. PMID- 12911447 TI - Comparison of the response to ovarian stimulation between women with one ovary and those with two ovaries, in a program of ICSI/ET. AB - BACKGROUND: Women with a single ovary form a group of special interest in assisted reproduction. The aim of this study was to compare the outcome of intracytoplasmic sperm injection cycles between women with one and two ovaries. To our knowledge, this is the first study reporting the outcome of intracytoplasmic sperm injection cycles with women with a single ovary. METHODS: Sixty-three intracytoplasmic sperm injection cycles in 24 women with a single ovary were compared with 191 intracytoplasmic sperm injection cycles in 109 women with both ovaries. All cycles were stimulated with triptorelin-long protocol. RESULTS: Statistically significant differences were found in the following parameters between women with a single ovary and women with two ovaries: total number of administrated gonadotropins (63.22 +/- 45.03 vs. 44.72 +/- 21.92), number of follicles (8.29 +/- 5.02 vs. 14.45 +/- 7.94), estradiol peak levels (1695.05 +/- 1177.34 vs. 2728.51 +/- 1852.67), number of retrieved metaphase II oocytes (6.95 +/- 3.78 vs. 11.72 +/- 6.11) and number of 2 PN oocytes (4.07 +/- 2.85 vs. 6.53 +/- 4.14). There were no differences in duration of stimulation, number of transferred embryos, and cumulative embryo score. In the group of women with a single ovary 14 pregnancies were achieved (22.2%), whereas 33 pregnancies were achieved in the group of women with two ovaries (17.28%). CONCLUSIONS: Women with a single ovary have a decreased response to external ovarian stimulation than women with both ovaries, but they present a higher, although not statistically significant, pregnancy rate. PMID- 12911448 TI - Obstetric and neonatal risk of pregnancies after assisted reproductive technology: a matched control study. AB - BACKGROUND: The aim of the study was to evaluate the obstetric and neonatal outcome of pregnancies after assisted reproduction technology (ART) in comparison with matched controls from spontaneous pregnancies. METHODS: A total of 12 920 deliveries at the Department of Obstetrics and Gynecology, University of Szeged, from 1 January 1995 to 31 December 2001 were subjected to retrospective analysis. Two hundred and eighty-four singleton, 75 twin and 17 triplet pregnancies after ovulation induction (n = 114; 30.3%), intrauterine insemination (n = 33; 8.8%) and in vitro fertilization (n = 229; 60.9%) were evaluated. The pregnancy outcome of the singleton and twin pregnancies was compared with that for controls matched with regard to age, gravidity and parity and previous obstetric outcome after spontaneous pregnancies. RESULTS: Twenty-four percent of the assisted reproductive pregnancies were multiple pregnancies. The incidences of singleton intrauterine growth retardation (IUGR) and preterm birth were reasonably similar to those among the controls (IUGR: 6.3% vs. 4.2%; preterm births: 13.0% vs. 9.9%, for the cases and the controls, respectively). As compared with the controls, there was an increased incidence of cesarean section among the singleton (41.2% vs. 34.5%, p = 0.12; OR 1.33; 95% CI 0.95-1.87) and twin assisted reproduction pregnancies (66.7% vs. 60.0%), but without significant differences. CONCLUSIONS: Increased obstetric risk could be observed concerning threatened preterm delivery and cesarean section rate in the study group. The perinatal outcome of singleton and twin pregnancies following assisted reproductive techniques is comparable with that of spontaneously conceived, matched pregnancies. PMID- 12911450 TI - Pitfalls in diagnosis of interstitial pregnancy. AB - BACKGROUND: To determine the incidence and reasons for misdiagnosis in interstitial ectopic pregnancy. METHODS: We conducted a retrospective study from 1990 to 2001. Women with interstitial pregnancy were identified and their case records retrieved and reviewed. Reasons for delay in diagnosis and associated morbidity were recorded. RESULTS: Thirty-six women were diagnosed as having interstitial pregnancy during the study period. Initial diagnosis was incorrect in 15 cases (41.7%). In 14 cases, the interstitial pregnancies were mistaken as intrauterine pregnancy. These misdiagnoses resulted in six inappropriate surgical procedures (evacuation of uterus) being performed and led to rupture of interstitial pregnancy in eight women. In two women, the interstitial pregnancy was mistaken as normal intrauterine pregnancy while the uterus itself was thought to be a cervical fibroid. In both cases, the interstitial pregnancies ruptured at 18-20 weeks of gestation. CONCLUSION: Despite advances in sonographic skills and equipment and the availability of beta-human chorionic gonadotropin (hCG) monitoring, misdiagnosis of interstitial pregnancy still occurs frequently. Clinicians should be aware of the limitations of various investigations and maintain a high index of suspicion. PMID- 12911449 TI - Effect of combined oral estrogen/progestogen preparation (Kliogest) on bone mineral density, plasma lipids and postmenopausal symptoms in HRT-naive Thai women. AB - BACKGROUND: Kliogest is commonly prescribed for the relief of acute postmenopausal symptoms and prevention of postmenopausal bone loss. However, there have been few data on its effect in Asian women. METHODS: This 1-year, single-center, randomized, double-blind and placebo-controlled study evaluated the efficacy and safety of Kliogest in hormone replacement therapy (HRT)-naive Thai women. The subjects were 120 healthy Thai women aged between 45 and 65 years, with intact uterus, and who had been amenorrheic for at least 1 year. RESULTS: Kliogest increased spine (+ 6%, p < 0.01) and hip (+2%, p < 0.01) bone mineral density (BMD), and lowered plasma total cholesterol (TC) (-16%, p < 0.05) and low density lipoprotein cholesterol (LDL-C) (-16%, p < 0.05) concentrations. However, Kliogest also resulted in a decrease in high density lipoprotein cholesterol (HDL-C) concentration (-18%, p < 0.05). Compared to placebo, the reduction in menopausal symptoms by Kliogest was not statistically significant. The frequency and severity of treatment-related uterine bleeding decreased with the duration of Kliogest treatment. Furthermore, there was a fairly strong relationship between the change in serum estrone concentration and the average monthly weighted bleeding scores over the first 6 months (Spearman's correlation r = 0.54; p < 0.001), which became weaker over the entire treatment period (Spearman's correlation r = 0.27; p < 0.01). Although there was a small to moderate relationship between baseline estrone concentration and both lumbar (r = 0.23, p < 0.02) and hip (r = 0.20, p < 0.05) BMD, there was no significant relationship between Kliogest-induced change in estrone concentration and change in lumbar and hip BMD. CONCLUSIONS: Continuous treatment with Kliogest for 1 year reversed the potential postmenopausal bone loss in HRT-naive Thai postmenopausal women. However, its effect on cardiovascular risk is uncertain. Furthermore, Kliogest is safe but appears to have no significant effect on climacteric symptoms in the patients in the present study. PMID- 12911451 TI - A randomized trial comparing changes in psychological well-being and sexuality after laparoscopic and abdominal hysterectomy. AB - BACKGROUND: To evaluate changes in psychological well-being and sexuality 1 year after laparoscopic and abdominal hysterectomy. MATERIAL AND METHODS: Seventy-four women scheduled for abdominal hysterectomy due to benign disorders and assessed as not suitable for vaginal hysterectomy were randomized to abdominal hysterectomy (n = 38) or laparoscopic hysterectomy (n = 36). The Psychological General Well-Being (PGWB) Index was used to evaluate changes in psychological well-being and the McCoy scale was used to evaluate changes in sexuality. The differences in outcome before and 1 year after the hysterectomy were calculated for each individual and the changes were compared within and between the two patient groups. RESULTS: There were no significant differences between the two patient groups with respect to changes in psychological well-being and sexuality 1 year after surgery. CONCLUSIONS: This study implies that psychological well being and sexuality after hysterectomy are not influenced by surgical technique. PMID- 12911452 TI - A study on the microheterogeneity of alpha-fetoproteins produced by yolk sac and germ cell tumors. AB - BACKGROUND: It is generally believed that the lower the grade of differentiation of glycoprotein-producing cells, the more often modification by bisecting N acetylglucosamine (GlcNAc) or fucose (Fuc) at the sugar chain of the glycoprotein or increase in branching of side chains occurs. We examined the characteristics of the alpha-fetoprotein (AFP) sugar chain stored in amniotic and exocoelomic fluid during 5-9 weeks of gestation and analyzed serum-derived AFP of patients with germ cell tumors. METHODS: Total AFP concentrations in embryonic fluid at 5 9 weeks of gestation (n = 11) and serum of patients with germ cell tumors (n = 7) were measured using a radioimmunoassay (RIA) method. The percentages of AFPs reactive with Lens culinaris agglutinin (LCA), concanavalin A (Con A), erythroagglutinating phytohemagglutinin-E4 (E-PHA) and Ricinus communis agglutinin-120 (RCA 120) were obtained by lectin-affinity electrophoresis coupled with antibody-affinity blotting. RESULTS: It was revealed that at 5-9 weeks of gestation, AFP variants that had been modified by the Fuc residue, which bound to the GlcNAc residue at the reducing end of the sugar chain, and bisecting GlcNAc residues gradually decreased as pregnancy advanced; however, the presence of N acetylneuraminic acid (Neu5Ac) at the nonreducing ends changed little. CONCLUSIONS: It appears very likely that the changes in the relative amounts of AFP variants in the embryonic fluid during early pregnancy were due to differentiation of the yolk sac. The grade of differentiation of yolk sac tumors was very similar to that of the normal yolk sac at around 6 weeks of gestation. PMID- 12911453 TI - Detection of cervical intraepithelial neoplasia in women with atypical squamous or glandular cells of undetermined significance cytology: a prospective study. AB - BACKGROUND: (1) To assess the prevalence of histologically confirmed cervical intraepithelial neoplasia in patients with cervical smears diagnosed as atypical squamous or glandular cells of undetermined significance. (2) To evaluate the role of colposcopy and the presence of human papillomavirus in detecting underlying cervical intraepithelial neoplasia. MATERIALS AND METHODS: In this prospective cohort, 148 women with atypical squamous or glandular cells of undetermined significance were evaluated by colposcopy, histological sampling, and human papillomavirus deoxyribonucleic acid testing. RESULTS: Histological diagnosis of >/= cervical intraepithelial neoplasia II was found in 10/148 women. Women with a histological >/= cervical intraepithelial neoplasia II had a higher prevalence of >/= two abnormal quadrants (90% vs. 38%/= cervical intraepithelial neoplasia II. PMID- 12911455 TI - Usefulness of language and memory testing during intracarotid amobarbital testing: observations from an fMRI study. AB - BACKGROUND: Several procedures for testing language lateralization and memory function exist during the intracarotid amobarbital test (IAT). The use of functional magnetic resonance imaging (fMRI) gives the opportunity to assess the validity of some of these procedures, or at least to inspect the neuronal correlates. A comprehensive fMRI protocol was tested, aimed at addressing aspects of lateralization of language, as well as testing memory in relation to activation of mesiotemporal regions. Here we report observations with possible consequences for the current IAT procedures. MATERIALS AND METHODS: The protocol consisted of three language tasks (overt naming, semantic decision and silent word generation) and two memory tasks (encoding and retrieving visual scenes). The paradigms used a block-related procedure in nine right-handed normal volunteers. During the procedure dynamic weighted full brain images were acquired which are sensitive to the blood oxygenation activation effect. RESULTS: Encoding showed symmetrical bilateral activation in the mesiotemporal regions, specifically the hippocampus, parahippocampal gyrus and fusiform gyrus. With a retrieval task activation of the mesiotemporal areas was restricted to the posterior hippocampal area. Overt object naming showed results, similar to encoding tasks with bilateral activation of hippocampal areas. Silent word generation showed much stronger ability to lateralize than the other two language related tasks and especially object naming. CONCLUSION: Activation revealed by fMRI activation shows that IAT procedures, using active semantic language processing or comprehensive procedures with multiple language tasks have the highest guarantee for individual activation lateralization. Simple object naming does not guarantee a lateralized language fMRI activation pattern. Of the different memory procedures during IAT, the procedures (Interview and the Montreal) demanding encoding processing will be related to larger areas of bilateral hippocampal activation than procedures (Seattle) exclusively requiring retrieval. Moreover, tasks using recognition of previously presented language items (naming objects) are equally effective for assessing hippocampal activation compared with presenting separate memory items. PMID- 12911456 TI - Does imprint cytology of brain tumours improve intraoperative diagnoses? AB - OBJECTIVES: To evaluate the diagnostic accuracy using frozen sections only and a combination of imprint cytology and frozen sections. MATERIAL AND METHODS: After introduction of imprint cytology as a supplement to frozen sections in 1999, 153 patients with brain tumours underwent stereotactic or open surgery. An equal number of cases prior to 1999 were chosen for comparison. Intraoperative diagnoses were compared with final diagnoses based on paraffin sections of the same tissue samples. The number of delayed intraoperative diagnoses was noted in each patient group. RESULTS: The combined use of the two techniques improved intraoperative diagnostic accuracy from 87 to 91% while the delayed intraoperative diagnoses were significantly reduced from 30 to 8. The choice of surgical procedure did not affect the outcome of the pathological investigations. CONCLUSION: A combination of frozen sections and imprints significantly reduced the number of delayed intraoperative diagnoses. Intraoperative diagnostic accuracy was improved, although not to a statistically significant level. Choice of surgical procedure did not affect the diagnostic outcome. PMID- 12911457 TI - Axonal and demyelinating forms of the MPZ Thr124Met mutation. AB - OBJECTIVE: We report on a Japanese family with Charcot Marie Tooth disease (CMT) with the Thr124Met mutation in the peripheral myelin protein zero (MPZ) gene. MATERIAL AND METHODS: Based on the clinical study, we investigated MPZ gene by direct sequence analysis and polymerase chain reaction restriction fragment length polymorphism analysis. RESULTS: Genotyping of four symptomatic family members showed that one family member with severe disease symptoms was homozygous, while the other three were heterozygous. The heterozygous cases were clinicopathologically determined to be the axonal type, which is characterized by late-onset and slow progression associated with Adie's pupil and deafness. The homozygous case was the demyelinating type, which showed earlier onset, rapid progression, sural nerve demyelination, and cranial nerve demyelination at autopsy. CONCLUSIONS: We suggest that axonal and demyelinating forms of CMT are not two distinct classes, but rather parts of a spectrum of genotypically related conditions, particularly with some MPZ mutations. PMID- 12911458 TI - Prognostic significance of SSEP, BAEP and serum S-100B monitoring after aneurysm surgery. AB - OBJECTIVE: Changes in evoked potentials (EPs) and increased levels of S-100B protein were used to identify cerebral ischemia or glial damage and to predict neurological outcome in aneurysm patients. MATERIAL AND METHODS: Somatosensory evoked potentials and Brainstem auditory-evoked potentials, and serum S-100B protein were simultaneously investigated pre- and postoperatively over a period of 10 days in 43 patients with 47 aneurysms (six in the posterior fossa). RESULTS: The EP scores showed a strong correlation with the clinical outcome. Sensitivity was 73%, and specificity 81%. Pathological S-100B levels >0.5 mg/l were equal in predictive values (correct positive eight, false positive six, correct negative 26, false negative three). Initially increased S-100B levels, long-lasting S-100B elevation, and secondary increasing S-100B values correlated with an unfavorable outcome. High peak S-100B values correlated with bad EP scores at discharge. EP deterioration was the first indicator anticipating S-100B elevation and clinical deterioration in five patients. There was a good correlation between pathological S-100B values or EP findings and infarction on CT scan. CONCLUSIONS: Both EPs and S-100B protein showed a comparable high predictive value for outcome. S-100B reflects the extent of primary brain damage after subarachnoid hemorrhage and time course of ongoing secondary brain damage. Evoked potentials assess the functional integrity and tended to react earlier than S-100B protein before definitive structural damage occurred. PMID- 12911459 TI - Complex spinal reflexes during transcranial Doppler ultrasound examination for the confirmation of brain death. AB - OBJECTIVE: Complex sets of movements of the extremities can be seen in patients with brain death (BD), and are typically observed during apnea testing or removal of ventilatory support (also called the Lazarus sign). We here describe brain dead patients who had not shown previous movements, even during apnea testing, but presented complex spinal reflexes during transcranial Doppler (TCD) examination elicited by neck flexion. METHODS: We performed a prospective TCD study of patients with the clinical diagnosis of BD. RESULTS: Four (2.5%) of 161 brain-dead patients presented complex spinal reflexes exclusively during TCD examination. TCD showed vertebro-basilar circulatory arrest in all four. Their systolic blood pressure was significantly lower than that of brain-dead patients not presenting movements during TCD examination. CONCLUSIONS: Hypotension and mechanical stimulation play a role in the pathophysiology of complex spinal reflexes present in BD, which are not exclusively seen in terminal hypoxia. Intensive care personnel and neurologists who perform TCD to confirm BD should be aware of these movements. PMID- 12911460 TI - Predicting the course of myasthenic weakness following double filtration plasmapheresis. AB - OBJECTIVE: To evaluate the clinical course of patients with myasthenia gravis (MG) up to 3 months after double filtration plasmapheresis (DFP). MATERIAL AND METHODS: We recorded the MG score and measured the level of acetylcholine receptor antibody (AchRAb) at baseline and day 1 (D1), week 1 (W1), 1 month (M1), 2 (M2) and 3 months (M3) after DFP in 16 MG patients. Based on the difference in score during follow-up, we divided our patients into clinical improvement (CI) and clinical worsening (CW) groups. RESULTS: The MG score decreased in all courses from a mean of 8.1 at baseline to 5.6 at D1, and to 4.7, 4.0, 3.8, and 3.7 at W1, M1, M2, and M3, respectively. In the CW group, AchRAb levels were significantly higher at M1 (P = 0.022). The AchRAb level at W1 correlated significantly with the MG score at M3 (P = 0.027) and the changes of MG score from W1 to M1 (P = 0.029). The ratio of AchRAb levels of M1 to W1 correlated well with MG score at W1 (P = 0.032), at M3 (P = 0.001), and the changes of MG score from W1 to M1 (P = 0.004). CONCLUSION: Excessive rebounds of AchRAb level at W1 may suggest clinical worsening and further increases in AchRAb level at M1 predict poorer outcome after DFP. PMID- 12911461 TI - NMDA-mediated mechanisms in cortical excitability changes after limb amputation. AB - OBJECTIVES: The aim of our study was to determine the role of N-methyl-d aspartate (NMDA)-mediated mechanisms in cortical excitability changes after limb amputation, and their possible relationship to phantom pain. MATERIALS AND METHODS: Sixteen upper limb amputees who were suffering from chronic phantom pain received the NMDA-antagonist memantine or placebo for 3 weeks. Intracortical inhibition (ICI) and intracortical facilitation (ICF) were determined at baseline and on day 21 using transcranial magnetic stimulation. Simultaneously, phantom pain intensity was assessed. RESULTS: Memantine reduced ICF and enhanced ICI to roughly the same extent as seen in healthy subjects in a previous study. These changes were not correlated to the reduction of phantom pain. CONCLUSION: We therefore conclude that NMDA-mediated mechanisms influence changes of ICI and ICF occurring after limb amputation. However, our results suggest that these cortical excitability changes and phantom pain are independent of each other. PMID- 12911462 TI - Thrombolytic and anticoagulation treatment in a rat embolic stroke model. AB - OBJECTIVES: The effects of pentasaccharide (PENTA), given alone or combined with thrombolysis using recombinant tissue plasminogen activator (rt-PA), on infarct size and clinical outcome were evaluated in a rat embolic stroke model. MATERIALS AND METHODS: Ninety-two rats were embolized unilaterally and assigned to: (i). controls, (ii). rt-PA 6 mg/kg, (iii). PENTA 0.5 mg/kg, (iv). PENTA 0.5 mg/kg and rt-PA 6 mg/kg. After 2 days animals were killed, the brains removed and evaluated microscopically. RESULTS: The median infarct size measured in percentage of the affected hemisphere was 25% in the control group, 4% (P < 0.01, Mann Whitney) in group 2, 19% (n.s.) in group 3, and 10% (P < 0.05) in group 4. rt-PA, and rt-PA combined with PENTA also promoted functional recovery. CONCLUSION: The present study found no effect of 0.5 mg/kg PENTA treatment. Compared with rt-PA treatment alone, 0.5 mg/kg PENTA alone or combined with rt-PA did not significantly increase mortality or tendency for hemorrhage. PMID- 12911463 TI - Devic's neuromyelitis optica: study of nine cases. AB - OBJECTIVE: Multiple sclerosis (MS) is by far the most popular diagnosis for patients with multifocal neurological disease. Owing to demyelinating inflammatory non-necrotic plaques of the white matter, MS can give remitting symptoms of virtually every part of the central nervous system. Corticosteroids are usually helpful. Devic's neuromyelitis optica (DNMO) is a neurological disease involving only the optic nerves and the spinal cord, where demyelination evolves towards necrosis and atrophy; the prognosis is poor and no satisfactory treatment is known. The objectives of this study are to describe clinical, biological, pathological and radiological data of patients with DNMO and to differentiate DNMO from MS. MATERIAL AND METHODS: We studied the files of 14 patients diagnosed with possible DNMO in three French hospitals between 1980 and 1999 and reviewed the literature. RESULTS: Nine patients were included as definite DNMO. Five were excluded because they did not fulfil the diagnostic criteria. For the nine patients with definite DNMO, DNMO was either monophasic or multiphasic. The prognosis was generally poor: two patients died and five others developed severe disability such as blindness, para or quadriplegia or both. Cerebrospinal fluid study and neuroimaging were essential to confirm the diagnosis of DNMO. Various immunosuppressive treatments generally failed to benefit the patients. CONCLUSION: In the literature (as well as our 14 initial patients) only a few cases of patients described as suffering from DNMO fulfilled the diagnostic criteria. The others showed evidence that another disease like MS was involved. We stress that inclusion and exclusion criteria have to be kept in mind to differentiate clearly DNMO from MS and other central nervous system white matter diseases. PMID- 12911464 TI - Representational cortical plasticity associated with brain tumours: evidence from laser-induced interstitial thermotherapy. AB - OBJECTIVES: To test the hypothesis that cortical plasticity related to destructive tumour growth is functionally relevant. This hypothesis predicts that function is dependent on the intactness of tissue surrounding the tumour. MATERIAL AND METHODS: Eight patients underwent laser-induced interstitial thermotherapy (LITT) for minimally invasive palliative treatment of brain tumours located in eloquent frontal motor regions including the primary motor cortex. A multimodal approach was used to assess the functional outcome of patients after LITT in detail. RESULTS: Following LITT, motor function deteriorated in four patients. In three of these four patients the LITT-induced lesion involved minimal parts of adjacent non-tumorous tissue. By contrast, the other four patients whose LITT-induced signal changes were confined to the tumour, showed no functional deficits. CONCLUSION: These findings support the idea that peri tumorous neuronal circuitry in motor competent areas may permanently take over those functions that were formerly represented in the neuronal tissue destroyed by the tumour. PMID- 12911465 TI - Distal myasthenia gravis frequency and clinical course in a large prospective series. AB - OBJECTIVES: In Myasthenia gravis (MG) proximal limb, ocular and/or bulbar muscles are most commonly affected, whereas distal extremity muscles are typically spared. The aim of the current study was to assess the frequency of primarily distal MG in the Tyrol and to describe its clinical peculiarities. MATERIAL AND METHODS: Over the past 20 years 84 patients with MG have undergone follow-up at the Department of Neurology of Innsbruck University. Types of presentation, clinical course and treatment response were followed over a period of 20 years (1980-2000). RESULTS: Six of 84 MG patients showed a predominance of muscle weakness and fatigability in distal limb muscles (two at presentation, four over the later course of the illness). There was no difference between distal MG and MG with a more typical distribution of muscle weakness regarding age, gender and response to therapy. CONCLUSIONS: The case series indicates that predominantly distal presentations of otherwise typical MG are more frequent than generally assumed and should be considered in the differential diagnosis of diseases with distal limb weakness. PMID- 12911466 TI - Bedside biochemical monitoring of the penumbra zone surrounding an evacuated acute subdural haematoma. AB - We describe a penumbra zone with increased biochemical vulnerability in cerebral cortex underlying an evacuated acute subdural haematoma. Two microdialysis catheters were placed in this zone and one catheter was placed in the opposite, less injured hemisphere. The microdialysis perfusates were analysed bedside for glucose, pyruvate, lactate, glutamate, and glycerol. In the penumbra zone, but not in the opposite hemisphere, energy metabolism was seriously disturbed with signs of cell membrane degradation. During an adverse event (decrease in haemoglobin level, systemic blood pressure and cerebral perfusion pressure) the perturbation of energy metabolism increased in this zone. Energy metabolism recovered and the signs of cell membrane degradation disappeared after normalization of the physiological parameters. We use the term biochemical penumbra zone to describe an area with signs of energy failure and cell membrane degradation, which has a capacity to regain a normal metabolic pattern but also an increased vulnerability to secondary insults. PMID- 12911467 TI - Bobble-head doll syndrome: some atypical features with a new lesion and review of the literature. AB - Bobble-head doll syndrome is a rare and unique movement disorder encountered in children. It is characterized by continuous or episodic involuntary forward and backward and side to side movement of the head at the frequency of 2-3 Hz. Neuroimaging in most of the cases reveals third ventricular tumors, suprasellar arachnoid cysts, aqueductal stenosis and other lesions in the region of the third ventricle along with communicating hydrocephalus. In most of the circumstances, the problem starts in the first decade of life and diversion of cerebrospinal fluid by shunt operation is very often accompanied by dramatic improvement. We report one case where bobbing of the head started at around 12 years of age. Additionally, there was evidence of partial left abducens nerve palsy, tremor in the outstretched hands, difficulty in finger-nose test and tandem walking, hyperreflexia and extensor plantar response. He was unconscious on two occasions and there was evidence of gross hydrocephalus along with a thin membranous web, running transversely across the lower part of the aqueduct of Sylvius without any cerebrospinal fluid flow void. Ventriculo-peritoneal shunt abolished the abnormal movements. We propose that the aqueductal web was the offending agent for the pathogenesis of bobble-head doll syndrome in our case and this lesion has not been identified in the cases reported so far. Relevant literature in this regard has also been reviewed. PMID- 12911469 TI - Effect of maternal NG-nitro-l-arginine administration on fetal growth and hypoxia induced changes in newborn rats. AB - BACKGROUND: Nitric oxide (NO) inhibition with NG-nitro-l-arginine methyl ester (l NAME) in the last trimester of pregnancy caused intrauterine growth retardation and hind-limb disruptions in rats. In the present study, the effect of maternal NO inhibition with NG-nitro-l-arginine (l-NNA) on hypoxic newborn rats was investigated. METHODS: Timed-pregnant rats were obtained on gestational day 17. Four groups of rats were used: control, hypoxic, l-NNA and l-NNA + hypoxic groups. In the last two groups, l-NNA (2 mg/kg bolus, i.p.) was administered to the mothers of pups antenatally on 3 consecutive days. Hypoxia was induced in newborn rats by breathing of a mixture of 8% oxygen and 92% nitrogen for 3 h. Pups were then allowed to inhale normal atmospheric air for 30 min. All newborn rats were decapitated on the first day of life after hypoxia and reoxygenation. Brain, heart, lung, liver, kidney and intestinal tissues were studied biochemically. Hypoxia-induced biochemical changes were determined by measuring lipid peroxidation. Histopathologic examination of lung tissue was performed. RESULTS: Nitric oxide synthase inhibition in pregnancy did not cause fetal growth retardation. Hypoxia increased lipid peroxidation in all tissues except the heart; this increase was decreased by maternal l-NNA administration in brain, lung, liver and kidney tissues. However, lipid peroxidation was increased by NO synthase inhibition in the intestines. In the lungs, pulmonary hemorrhage was observed in the hypoxic group. Minimal pulmonary hemorrhage was detected in the l NNA and l-NNA + hypoxic groups. CONCLUSIONS: These data suggest that antenatal administration of an NO synthase inhibitor acts as both a destructive and protective agent in hypoxic newborn rats. PMID- 12911468 TI - Plasma TGFbeta1 in ALS patients. PMID- 12911470 TI - Neonatal clinical outcome after elective cesarean section before the onset of labor at the 37th and 38th week of gestation. AB - BACKGROUND: Although elective cesarean sections are often performed after the 37th week of gestation without any complicating factor that may influence the timing of delivery,there is a possibility that infants born in the 37th week of gestation,especially early in the 37th week of gestation, do not obtain a satisfactory clinical outcome due to premature birth. METHODS: The authors analyzed the clinical course during the neonatal period in 96 infants born in the 37th (n = 81)and 38th (n = 15) week of gestation by an elective cesarean section. Subjects were retrospectively divided into two groups: infants born in the first half of the 37th week of gestation (37+0-37+3)(n = 48), and infants born from the latter half of the 37th week of gestation (37+4-37+6)through the 38th week of gestation (n = 48). Twin pregnancies, pregnancy with placenta previa, and pregnancy of women who had a diversity of medical complications were excluded from the present study, because of the possibility that these conditions may have affected the infants' status. The incidence of infants who showed clinical symptoms during the neonatal period and who needed medical care was compared between the two groups. RESULTS: Of the 96 subjects, 25 infants(26.0%) had significant clinical symptoms. The incidence of breathing difficulty was significantly higher in the infant group born in the first half of the 37th week of gestation than in the latter group. CONCLUSIONS: An elective cesarean section before the onset of labor early in the 37th week of gestation should not be routinely undertaken. PMID- 12911471 TI - Congenital dilatation of the bile duct: changes in diagnostic tools over the past 19 years. AB - BACKGROUND: Because of recent improvements in diagnostic procedures, many cases of congenital dilatation of the bile duct (CDBD) have been diagnosed in early life. To determine the mode of presentation and changes in diagnostic tools, medical records of 34 children (25 girls, nine boys) with CDBD who were treated between 1982 and 2000 were reviewed. RESULTS: The age at presentation ranged from 0 to 16 years (median 4.9 +/- 0.7 years), with two patients being diagnosed using prenatal ultrasonography. Of the 32 patients diagnosed post-natally, 21 (66%) complained of abdominal pain. There were also 21 (66%) cases of vomiting. Sixteen patients (50%) experienced both these symptoms. Ten (31%) cases presented with jaundice and three (9%) presented with a palpable mass. Ultrasonography was used as a diagnostic tool in 29 cases (85%). The finding of a dilated common bile duct on abdominal ultrasonography was helpful for early diagnosis. Spiral computed tomography was also a useful and accurate imaging method for diagnosis. Anomalous pancreaticobiliary junction (PBJ) was demonstrated in 44% of patients with CDBD. Magnetic resonance cholangiopancreatography, which can be performed non invasively, is useful for the diagnosis and pre-operative assessment of CDBD in children. The average length of time from presentation to diagnosis could be shortened from 46 days (1982-1988) to 13 days (1995-2000). CONCLUSION: Recent improvements in diagnostic procedures make immediate, non-invasive diagnosis of CDBD and PBJ possible. PMID- 12911472 TI - Long-term outcome after sclerotherapy with or without a beta-blocker for variceal bleeding in children. AB - BACKGROUND: Esophageal variceal bleeding is a life-threatening complication of portal hypertension. Optimal treatment for the prophylaxis of variceal rebleeding in children has not yet been determined. In the present study, we aimed to compare the long-term efficacy of endoscopic sclerotherapy with or without oral beta-blocker therapy in the secondary prophylaxis of variceal bleeding. METHODS: Thirty-eight children who had undergone endoscopic sclerotherapy (EST) sessions for variceal bleeding in the Department of Pediatric Gastroenterology, Istanbul University Istanbul School of Medicine, were entered into this retrospective cohort study. Twenty patients (mean +/- SD age 7.0 +/- 2.7 years) had undergone only sclerotherapy sessions (SG), whereas 18 patients (mean age 6.8 +/- 3.4 years) had received oral propranolol (1-2 mg/kg per day) additionally for 2 years (SPG). The number of patients with successful obliteration, the time required for obliteration and variceal recurrence rate were analyzed as primary indicators of the effectiveness of therapy. RESULTS: Variceal obliteration was achieved in 16 of 20 patients (80%) in the SG group and in 16 of 18 patients (88%) in the SPG group. Time required for variceal obliteration was significantly shorter in the SPG group compared with the SG group (4.1 +/- 1.4 vs 3.2 +/- 0.9 months; P < 0.05). The variceal recurrence rate was 65 and 38.8% in the SG and SPG groups, respectively. Compared with the SG group, less variceal rebleeding was observed during EST in the SPG group (25 vs 16.6%, respectively).However, these differences were not statistically significant. CONCLUSIONS: Endoscopic sclerotherapy combined with oral propranolol treatment shortens the time required for variceal obliteration. However, the other indicators of treatment effectiveness are not influenced statistically by the addition of propranolol to the treatment regimen. Randomized prospective clinical studies in larger pediatric series are needed before offering a combination of EST with oral propranolol as the most rational approach in the secondary treatment of esophageal variceal bleeding in children. PMID- 12911474 TI - Feeding practice of infants and their correlates in urban areas of Beijing, China. AB - BACKGROUND: Optimal feeding practice in the first year of life is crucial for the survival and health of infants, and has long-term consequences in later life. However, non-optimal feeding practices exist widely. The present study aims to explore various constraints to optimal feeding practices in the first year of life of infants in urban areas of Beijing, China. METHODS: A cross-sectional study was conducted in urban areas of Beijing from 4 July to 20 August, 1998. Two hundred and fifty-one mothers of infants aged 6-12 months were chosen from six child health centers in three different urban districts in Beijing. A self administered structured questionnaire was used to collect data regarding feeding practices and potentially related factors. RESULTS: Feeding practice for most of the infants was in accordance with the national and international recommendations. However, the rate of incidence of exclusive breast-feeding at 3 months of age was lower than that recommended by the World Health Organisation (WHO) (55.8%), and the introduction of solid/semisolid food before 4 months of age was found in approximately 19.3% of the infants. Cow's milk was given to 21.2% of infants from 6 months of age as the sole source of milk or as a supplement. Maternal education level (OR = 2.44, 95% CI: 1.42-4.19, P < 0.05), employment (OR = 2.05, 95% CI: 1.13-3.74, P < 0.05) and antenatal nonexclusive breast-feeding plans (OR = 4.10, 95% CI: 2.24-7.50, P < 0.001) were found to be correlated to inappropriate feeding practices. CONCLUSIONS: The feeding practices for most of the urban infants was found to be in accordance with the Chinese government and WHO recommendations; however, non-optimal feeding practices presenting as the early cessation of breast-feeding and the introduction of solid/semisolid foods existed. Information regarding optimal feeding practices should be disseminated to mothers and medical professionals in China, to ensure optimal infant health. PMID- 12911473 TI - Celiac disease in children with Down syndrome: importance of follow-up and serologic screening. AB - BACKGROUND: Celiac disease, also known as gluten-sensitive enteropathy, is a chronic inflammation disease of the small intestinal mucosa. Detection of Ig-A antigliadin antibodies (AGA) and antiendomysial antibodies (EMA) in serum is important in the diagnosis and screening for celiac disease. Antiendomysial antibodies have greater sensitivity compared to antigliadin antibodies. It has been reported that the prevalence of celiac disease is higher in children with Down syndrome than the other autoimmune conditions. The aim of the present study was to investigate the incidence of celiac disease in children with Down syndrome, to assess the availability of Ig-A AGA and EMA for serologic screening, and to highlight the importance of follow-up for children with Down syndrome. METHODS: Forty-seven children with Down syndrome without known celiac disease were tested for total blood count, thyroid function tests, immunoglobulin values, Ig-A AGA and EMA. Duodenal biopsy was performed on eight patients who showed at least one serologically positive marker. RESULTS: The ages of the children with Down syndrome ranged from 2 to 18 years (30 boys/17 girls). The mean age was 6.55 +/- 3.88. Total blood count and immunoglobulin values were normal. Eleven of the 47 patients (23.40%) were found to be serologically positive, 10 (21.28%) having antigliadin antibody concentrations above normal; and six (12.77%) being positive for antiendomysial antibody. In five patients (10.64%), both Ig-A AGA and EMA concentrations were high and positive. Duodenal biopsies of three of eight cases (37.50%) revealed villous atrophy, lymphocyte infiltration and crypt hyperplasia. Three cases with abnormal biopsy results (100%) were below the 10th percentile for weight and height. Hypo-thyroidism was detected in one of 11 cases where at least one serologic marker was positive. CONCLUSION: Children with Down syndrome should be carefully examined in their follow up, and celiac disease should be considered in cases with growth retardation. Ig-A antigliadin antibodies and EMA are non-invasive, cheap and readily available serologic screening tests for celiac disease, and the positivity of both markers gives the most reliable result. PMID- 12911475 TI - Autonomic function in Kawasaki disease with myocardial infarction: usefulness of monitoring heart rate variability. AB - BACKGROUND: Despite anticoagulant therapy, many patients with Kawasaki disease and giant coronary artery aneurysm develop myocardial infarction. These patients have a high risk of sudden death, but the etiology is not clear. We studied autonomic function and the possibility of malignant ventricular arrhythmia through heart rate variability. METHODS: We studied six Kawasaki disease patients with myocardial infarction and 16 normal controls. Heart rate variability was investigated using a 24 h electrocardiogram. We assessed the standard deviation from the mean of the normal R-R intervals (SDNN), the proportion of adjacent R-R intervals with a difference greater than 50 msec (pNN50) and the root-mean square of successive R-R differences as time-domain analysis (rMSSD). We assessed very low-frequency power, low-frequency power (LF), high-frequency power (HF) and the LF/HF ratio in frequency-domain analysis. RESULTS: There was no significant difference in SDNN, but there was a significant difference in pNN50 and rMSSD. Patients with Kawasaki disease showed lower HF and higher LF/HF than normal controls. CONCLUSION: Our findings suggest that patients with Kawasaki disease and myocardial infarction show decreased vagal activity, which could cause malignant arrhythmia. PMID- 12911476 TI - Case-control study of giant coronary aneurysms due to Kawasaki disease. AB - BACKGROUND: Epidemiologic features of the formation of giant coronary aneurysm due to Kawasaki disease and its risk factors are not still clear. METHODS: One hundred and five patients with giant coronary aneurysms were selected from the 15th and 16th nationwide surveys of Kawasaki disease in Japan. With all other patients reported from the same hospital as a control group, odds ratios (OR) were calculated for some potential risk factors. RESULTS: Some factors affecting the formation of giant coronary aneurysms were revealed. Of these factors, the following were considered to be useful for the treatment of patients with Kawasaki disease: male sex (OR compared with females = 1.99), 0 years old (OR compared with 1-2 years = 1.71), visiting the hospital on 1-3 days of illness (OR compared to 4-6 days = 1.72), leukocyte count (OR with 1000 /mm3 increase = 0.56), neutrophil concentration among leukocyte (OR with 10% increase = 1.11), hemoglobin concentration (OR with 1 mg/dL increase = 0.73), alanine aminotransferase level (ALT; OR with 10 IU/dL increase = 1.02) and serum sodium level (OR with 1 mEq/L increase = 0.80). CONCLUSIONS: The observation of 105 cases with giant coronary aneurysms due to Kawasaki disease reported in the nationwide survey provides information about some risk factors and other considerations about the aneurysms. PMID- 12911478 TI - Serial changes of plasma nitrate in the acute phase of Kawasaki disease. AB - BACKGROUND: Endogenous nitric oxide (NO) production increases with clinical conditions associated with immune stimulation. In Kawasaki disease (KD), various cytokines play a role in inflammatory reactions in the cardiovascular system. The authors hypothesized that elevated concentrations of nitrate was related to the severity of vasculitis. The aim of the present study was to evaluate serial changes of plasma nitrate concentrations in the acute phase of KD and to consider how NO is related to the inflammatory process of KD and to the coronary artery lesion (CAL). METHODS: Thirty patients with KD and 20 age-matched healthy controls were enrolled in the present study. Blood samples were obtained weekly for the first and second months. The patients were divided into two groups: one with CAL (n = 11) and another without CAL (n = 19). Plasma nitrate was measured by high-performance liquid chromatography. RESULTS: In both groups, plasma nitrate increased remarkably from the first week to the third week. Peak concentrations of nitrate (mean +/- SD, micro mol/L) in each group were as follows: 56.9 +/- 23.8 in the CAL(+) group and 68.2 +/- 33.8 in the CAL(-) group. Plasma nitrate decreased from the third week to the second month but was still elevated in both groups in comparison with the age-matched healthy controls. There was no correlation between plasma nitrates and white blood cell count or C reactive protein, respectively (r = 0.013, 0.075). CONCLUSIONS: The results suggest that NO production may not be related to the severity of vascular inflammation and that elevated nitrate during the first month of illness may not be associated with a higher risk of CAL. PMID- 12911477 TI - Echocardiographic evaluation of wheelchair-bound basketball players. AB - BACKGROUND: Cardiopulmonary function in sedentary men whose lower limbs have been immobilized for years has been shown to be markedly lower than normal. However, the cardiopulmonary function of paraplegics who regularly activate their upper limps and trunk has been suggested to be almost normal in a few studies. The purpose of the present study was to evaluate the left ventricular dimensions, left ventricular mass, systolic and diastolic function in adolescent wheelchair bound basketball players using echocardio-graphy, and to compare the results with those of sedentary adolescents unable to use their lower extremities and the results of able bodied controls. METHODS: The study group consisted of 22 male adolescent high school students who were unable to use their lower extremities: 11 were members of a high school basketball team who had been regularly playing basketball for at least 2 years, and 11 were sedentary adolescents none of whom was engaged in any kind of routine training program. The control group consisted of 11 healthy able-bodied male adolescents of similar age. RESULTS: There were no significant differences in left ventricular dimensions and wall thickness, aortic root, left atrium diameters, or left ventricular filling characteristics between the three groups. Wheelchair-bound basketball players had increased left ventricular ejection fraction and shortening fraction compared with the sedentary unable-bodied individuals. Although left ventricular ejection fractions were significantly lower than in normal adolescents, all ejection fraction values except one were within the normal limits in the unable-bodied basketball players. CONCLUSION: The results of the present study suggest that an upper extremity exercise program and sports such as basketball can improve the cardiac functions and additional echocardiographic functions of people unable to use their lower extremities, potentially to normal levels. PMID- 12911479 TI - Effects of passive smoking on lung function in children. AB - BACKGROUND: Passive smoking can have significant effects on lung function with reductions in forced vital capacity (FVC), forced expiratory volume in one second (FEV1) and forced mid-expiratory flow rate (FEF25-75%) of between 5 and 10%. METHODS: Fifty non-smoking children aged 6-15 years, who had no history of asthma or atopy and no parental history of atopy, were assessed with respect to their lung functions (FEV1, FEV1/FVC, FEF25-75%). RESULTS: Thirty-three of these children were being exposed to environmental tobacco smoke inside their homes, while 17 children were not exposed. In the 'passive smoker' group the FEV1, FEV1/FVC and FEF25-75% values were found to be significantly lower than the non smoker control group's values (P = 0.0080, 0.0228 and 0.0003, respectively). The decrease in FEF25-75% was significantly correlated inversely with the number of cigarettes smoked per day (P = 0.0261). CONCLUSION: There is sufficient evidence to support the notion that environmental tobacco smoke is a serious health burden for children. Considering that recent studies suggest that up to 70% of children grow up in homes with at least one smoker, every effort should be made to reduce these children's exposure to environmental tobacco smoke and to give them a chance to grow up in a more healthy environment. PMID- 12911480 TI - Risk factors of status epilepticus in children. AB - BACKGROUND: Although there is abundant literature about the morbidity and mortality rates of status epilepticus (SE), little is known about the risk factors of this medical emergency. The aim of the present study is to assess the risk factors of SE in children. METHODS: The authors reviewed the medical records of 83 patients admitted to the Pediatric Neurology Unit of Trakya University Hospital, Edirne, Turkey from January 1994 to December 2001 with the diagnosis of SE. Eighty-three patients were compared with 166 controls who were admitted to the same unit due to non-status epilepticus (non-SE) seizure. RESULTS: The univariate analysis demonstrated that SE episodes were significantly associated with a history of birth asphyxia, neonatal seizure, discontinuation of antiepileptic medication, epilepsy, partial seizure evolving to secondary generalized seizures, myoclonic seizure, generalized abnormalities in the neurological examination, neuromotor retardation, generalized background abnormalities on electroencephalogram (EEG), generalized abnormalities on neuroimaging and polypharmacy than non-SE episodes. Logistic regression was used to test the independence of these parameters as predictors of SE risk. Four parameters emerged as significant independent predictors of SE in children in multiple logistic regression: polypharmacy (Odds ratio (OR) 5.17, P = 0.0004), discontinuation of antiepileptic medication (OR 4.04, P = 0.0095), neuromotor retardation (OR 4.03, P = 0.0016) and generalized background abnormalities on EEG (OR 2.48, P = 0.0419). CONCLUSION: Polypharmacy, discontinuation of antiepileptic medication, neuromotor retardation and generalized background abnormalities on EEG are indicators in children of a higher risk of SE. PMID- 12911481 TI - Obesity and associated modifiable environmental factors in Iranian adolescents: Isfahan Healthy Heart Program - Heart Health Promotion from Childhood. AB - OBJECTIVE: To evaluate the prevalence of overweight and obesity among Iranian adolescents and their relationship with modifiable environmental factors. METHODS: The subjects of the present study were 1000 girls and 1000 boys, aged between 11 and 18 years selected by multistage random sampling, their parents (n = 2000) and their school staff (n = 500 subjects) in urban and rural areas of two provinces in Iran. Data concerning body mass index (BMI), nutrition and the physical activity of the subjects were analyzed by SPSSV10/Win software. RESULTS: The prevalence of 85th percentile 95th percentile in girls was significantly higher than boys (10.7 +/- 1.1 and 2.9 +/- 0.1%vs 7.4 +/- 0.9 and 1.9 +/- 0.1%, respectively; P < 0.05). The mean BMI value was significantly different between urban and rural areas (25.4 +/ 5.2 vs 23.2 +/- 7.1 kg/m2, respectively; P < 0.05). A BMI> 85th percentile was more prevalent in families with an average income than in high-income families (9.3 +/- 1.7 vs 7.2 +/- 1.4%, respectively; P < 0.05) and in those with lower educated mothers (9.2 +/- 2.1 vs 11.5 +/- 2.4 years of mothers education, respectively). The mean total energy intake was not different between overweight or obese and normal-weight subjects (1825 +/- 90 vs 1815 +/- 85 kCal, respectively; P > 0.05), but the percentage of energy derived from carbo-hydrates was significantly higher in the former group compared with the latter (69.4 vs 63.2%, respectively; P < 0.05). Regular extracurricular sports activities were significantly lower and the time spent watching tele-vision was significantly higher in overweight or obese than non-obese subjects (time spent watching telelvision: 300 +/- 20 vs 240 +/- 30 min/day, P < 0.05). A significant linear association was shown between the frequency of consumption of rice, bread, pasta, fast foods and fat/salty snacks and BMI (beta = 0.05-0.06; P < 0.05). A significant correlation was shown between BMI percentiles and serum triglyceride, high-density lipoprotein-cholesterol and systolic blood pressure (Pearson's r = 0.38, -0.32 and 0.47, respectively). CONCLUSIONS: Enhanced efforts to prevent and control overweight from childhood is a critical national priority, even in developing countries. To be successful, social, cultural and economic influences should be considered. PMID- 12911482 TI - Association between household conditions and diarrheal diseases among children in Turkey: a cohort study. AB - BACKGROUND: This aim of the present study was to estimate the frequency of diarrheal diseases among children as well as investigating the relationship between diarrheal diseases and household conditions and other factors. METHODS: The study was performed over 12 month period in Binevler Health Center, Gaziantep, Turkey. Five health stations were selected by using a sampling technique with probability proportional to size, and all households with children under the age of 5 years were visited. A questionnaire on diarrhea, household conditions, socioeconomic status of the family and individual characteristics of the children was applied to the mothers by doctors. Three composite indices, including household conditions, socioeconomic status of the family and individual status of the children, were prepared and a classification was made as good, mild or poor in order to better evaluate the results of the study. Daily recordings of diarrheal disease symptoms were made by the parents and these records were collected every month for 1 year. RESULTS: The annual mean incidence of diarrheal diseases was found to be 1.09 per child per year (median = 1). The mean was higher in children with poor household conditions (1.48 +/- 0.12) and with poor individual status (1.48 +/- 0.14), compared with good household conditions (0.76 +/- 0.07) and good individual status (0.71 +/- 0.08). The mean was also higher in children aged between 6 and 11 months (1.51 +/- 0.18), whose father graduated from primary school or lower (1.34 +/- 0.09) and with parents having no habit of washing their hands before taking care of the child (2.00 +/- 0.33). Person-month and case-month incidence rates of diarrhea were calculated to be 8.56 and 9.12%, respectively. CONCLUSIONS: Household conditions, individual status, age, education level of fathers and parents' habit of washing hands before taking care of the child were major factors affecting the incidence of diarrhea. PMID- 12911483 TI - Body mass index percentiles among adolescent girls living in Edirne, Turkey. AB - BACKGROUND: Body mass index (BMI) is the simplest way to measure obesity; therefore, it is chosen by many authorities as a screening method for adolescent obesity. Body mass index is positively correlated with the complications of childhood and adolescent obesity, such as hypercholesterolemia, insulin resistance, hyper-tension and long-term development of cardiovascular diseases. The aim of the present study was to produce percentile curves for bodyweight, height and BMI in a representative sample of adolescent girls living in urban and rural areas of Edirne, Turkey, and to compare these percentile curves with curves from other countries. METHODS: The present study was a cross-sectional study, including a representative sample of 1687 adolescent girls from rural and urban areas of Edirne, who were evaluated between May and July 2001. Bodyweight and height were measured using standard procedures. Body mass index (kg/m2) was calculated as the ratio of bodyweight to body height squared. Smoothed percentiles for these variables were calculated using polynominal regression models. Crude weight, height and BMI percentile values, as well as smoothed percentile curves are presented. RESULTS: Body mass index, weight and height reference curves for adolescent girls were produced. When we compared the BMI values of subjects in the present study with those of other countries, 85th and 95th percentiles of BMI in the present study were found to be generally lower than those for other ethnicities. CONCLUSION: Our findings show ethnic differences in BMI among adolescent girls. It will be usefull for each country to produce its own BMI percentiles. PMID- 12911484 TI - Spread of varicella-zoster virus DNA to the environment from varicella patients who were treated with oral acyclovir. AB - OBJECTIVE: The purpose of the present study was to determine the degree of spread of varicella-zoster virus to the environment (VZV) from varicella patients who received oral acyclovir treatment. METHODS: Over a period of 8 months, seven healthy children (two girls and five boys, 23-64 months of age) with varicella who visited Fujita Health University School of Medicine were treated with routine doses of oral acyclovir (ACV) for 5 days, commencing within 24 h after onset of the disease. Swab samples from the throats of patients and their family members as well as from air purifier filters in their houses were collected for 7 days as frequently as possible after starting treatment for the disease. The VZV DNA in the samples was identified by a sensitive polymerase chain reaction amplification assay. RESULTS: The VZV DNA was detected in 33-100% of throat swab samples from varicella patients by day 7 of the disease, in 17-32% of throat swab samples from family members by day 4 and in 29% of filters from air purifiers by day 3. CONCLUSIONS: The results suggest a broad spread of VZV, probably by the airborne route, from the patients with varicella even after the administration of oral ACV. PMID- 12911485 TI - Diphtheria immunization rates and the effect of several sociodemographic factors on immunization of children in eastern Turkey. AB - BACKGROUND: One of the aims of the World Health Organization is to eliminate diphtheria in all countries. Thus, it is important to achieve a high level of childhood immunization. The present study was performed to determine the level of child immunization against diphtheria and the effect of several sociodemographic factors on immunization of children aged 0-6 years. METHODS: Using the cluster sampling method, 767 healthy children were selected at random from three provinces in eastern Turkey. Children were categorized as completely vaccinated, incompletely vaccinated, unvaccinated or vaccination status unknown. If the child had no immunization card, he/she was categorized as vaccination status unknown. RESULTS: Of the 767 children, 72.3% were completely vaccinated, 18.3% were incompletely vaccinated and 5.7% were not vaccinated. Vaccination status was unknown for 3.7% of children. As the age of the children increased, the proportion of completely vaccinated children increased and the proportion of non vaccinated and incompletely vaccinated children decreased. The vaccination rates increased in parallel with the education level of the parents. While the vaccination rate was significantly correlated with the socioeconomic level of the families, it was inversely correlated with sibling number. CONCLUSIONS: The results of the present study show that the immunization status necessary for the elimination of diphtheria has not yet been reached in eastern Turkey. In immunization programs for children in eastern Turkey, priority should be given to increasing primary immunization levels to 90-95% with three or more doses of diphtheria toxoid. PMID- 12911486 TI - Rectal mucosal prolapse syndrome and a bidet. PMID- 12911487 TI - Rhabdomyosarcoma of the biliary tree. PMID- 12911488 TI - Psychosomatic reaction exhibiting hemorrhagic erythema and motorsensory disturbance. PMID- 12911489 TI - Transcatheter embolization of pulmonary collaterals using liquified gelatin. PMID- 12911490 TI - Adolescent with variant angina. PMID- 12911491 TI - Idiopathic arterial calcification in infancy with twin-twin transfusion syndrome. PMID- 12911492 TI - Cerebral vasculitis in Henoch-Schonlein purpura: MRI and MRA findings, treated with plasmapheresis alone. PMID- 12911493 TI - Mizoribine oral pulse therapy for a patient with severe lupus nephritis. PMID- 12911494 TI - Diagnostic appearance of abdominal roentgenogram in pica. PMID- 12911496 TI - Unexplained subdural hematoma in young children. PMID- 12911495 TI - First report of an outbreak of aseptic meningitis caused by echovirus type 13 in Japan. PMID- 12911498 TI - Moving forward in pediatric allergy & immunology. PMID- 12911500 TI - Eosinophil granule proteins in the assessment of airway inflammation in pediatric bronchial asthma. AB - Eosinophil granule proteins such as eosinophil cationic protein (ECP), eosinophil peroxidase (EPO) and eosinophil protein X (EPX) in serum and urine are indirect measures of eosinophil activity. The measures have been evaluated for prediction, diagnosis and monitoring of anti-inflammatory treatment modalities in children with asthma. Assessments in serum and urine are highly dependent on sampling procedures and must be performed under strictly controlled conditions using standardized sampling and laboratory procedures. The measures are influenced by circadian and seasonal variations. Measurement of the eosinophil granule proteins does not improve the predictive value of a family history of atopy. Due to insufficient sensitivity and specificity, the measures are not useful in the diagnosis of asthma in children, and the clinical use of eosinophil proteins in the individual child for assessment of asthma severity has not been sufficiently validated. Serum and urine eosinophil granule proteins, however, may be useful in extending our knowledge of suppressive effects on eosinophil activity of various doses, devices and administration regimens of inhaled glucocorticoids in children. Such evaluations may be performed in randomized, double-blind trials of well-defined age groups and they should include measures of compliance. One important aspect to look at would be the distinction between suppressive effects on eosinophil activity and clinically important anti-inflammatory effects. Considering the complexity of airway inflammation and the heterogeneity of childhood asthma, however, it may be too simplistic to look for a single measure of the inflammatory processes. In the future, perhaps, a combination of products of inflammatory cells may give more clinically relevant information with respect to prediction, diagnosis, monitoring and outcome of childhood asthma. PMID- 12911501 TI - The C-159T polymorphism in the CD14 promoter is associated with serum total IgE concentration in atopic Chinese children. AB - Activation of macrophages through CD14 by microbes is crucial in inducing immunity by type 1 T helper cells. A C-to-T polymorphism at position -159 of CD14 was associated with serum total IgE level in Caucasians but not in Japanese subjects. The objective of this study is to determine whether this polymorphic marker is associated with atopy and asthma phenotypes in Chinese children. Restriction fragment length polymorphism was used to characterize CD14/-159 genotypes. Microparticle immunoassay was used to measure serum total IgE level; fluorescent enzyme immunoassay was performed to measure serum concentrations of specific IgE to aeroallergens; and enzyme-linked immunosorbent assay was used to measure serum levels of soluble CD14 (sCD14). Lung function in asthmatics was assessed by spirometry. Two hundred and fifty-eight patients and 92 control children were recruited. Their mean serum total IgE concentrations were 331 and 74 kIU/l, respectively (p < 0.0001). Atopy, defined as the presence of at least one allergen-specific IgE in serum, was found in 220 (85%) patients and in 41 (45%) controls (p < 0.0001). Serum sCD14 levels were significantly associated with CD14/-159 genotypes (p = 0.004). Atopic subjects with CC genotype in CD14/ 159 had the highest serum total IgE levels compared with CT and TT genotypes, with the respective mean values being 661, 427 and 380 kIU/l (p = 0.015). Similarly, a higher proportion of subjects with CC genotype had increased serum total IgE concentration (p = 0.039). This polymorphic marker was not associated with asthma or aeroallergen sensitization in our cohort. Our results suggest that the C-159T of CD14 was associated with serum total IgE concentration in atopic Chinese children. PMID- 12911503 TI - Flow dependency and off-line measurement of exhaled NO in children. AB - Levels of exhaled nitric oxide (eNO) are flow-dependent, and the choice of an optimal flow rate for off-line and on-line eNO measurement has raised much debate. Recently, a flow rate of 50 ml/s was recommended, but children younger than 5-6 years are not capable of stabilizing their expiratory flow at low flow rates. The power of off-line eNO values to discriminate between normal and atopic children was therefore evaluated at different exhalation flow rates. At flow rates of both 8.3 ml/s and of 350 ml/s, children (8-12 years) sensitive to house dust mite have two-fold higher eNO values (p < 0.001) as compared with children lacking such a sensitivity. The power of eNO to discriminate between normal and atopic subjects was similar at the two flow rates (no difference in AUC of receiver operation curves, p = 0.89). All children from 4.5 to 5 years of age (n = 29) could perform a single off-line exhalation manoeuvre at high (>350 ml/s) but not at low (8.3 ml/s) flow rates. At high exhalation flow rate, eNO was 7.1 +/- 2.4 (mean +/- SD) median, 6.5 p.p.b. with a mean variation coefficient of 5.5%. Depending on their developmental level, about half of the younger children (35-46 months of age) could perform an off-line exhalation manoeuvre at high flow rate with good reproducibility (mean variation coefficient of 6.6%). It is concluded that an exhalation flow rate of 350 ml/s is feasible to determine off line eNO-values in children from 3.5 years of age, and that this high flow rate does not compromise the power of eNO to detect allergic disease. PMID- 12911502 TI - NOS1 polymorphism is associated with atopy but not exhaled nitric oxide levels in healthy children. AB - Exhaled nitric oxide (FENO) is raised in atopy. The mechanism for this is unclear. The aim of this study was to investigate whether the number of AAT repeats in intron 20 of the NOS1 gene, recently associated with variations in FENO in adults with asthma and cystic fibrosis, was associated with the raised FENO in healthy atopic children. Eighty-seven healthy children (44 girls, 42 atopic, age range 6-18 years) underwent measurements of FENO, spirometry, airway responsiveness and skin prick testing. Genotyping was carried out to determine the number of AAT repeats. There was no association between the number of AAT repeats and FENO in either the whole sample of healthy children (n = 87) or in the subsample of healthy atopics (n = 42). However, a greater number of atopic children had two high repeat alleles compared with non-atopic children (33.3% vs. 13.6%, respectively, p = 0.03). This suggests that variations in the NOS1 gene may contribute to atopy without this relationship being reflected by FENO. PMID- 12911504 TI - ISAAC-based asthma and atopic symptoms among Ha Noi school children. AB - Childhood asthma and atopy prevalence patterns in the developing world are only beginning to be defined. No such information exists for Vietnam. Estimates would assist in anticipating health service needs as well as add to the growing database on global patterns of atopy. To estimate the prevalence of atopic symptoms in school children in Ha Noi, Vietnam, a cross-sectional survey was conducted of children aged 5- to 11-years-old in two schools using the parent self-administered International Study of Asthma and Allergies in Childhood (ISAAC) questionnaire. The response rate was 66.4% (969 responses). The overall prevalence of selected symptoms were: 'ever wheezed' 24.9%, 'wheezed in past 12 months' 14.9%, 'ever had asthma' 12.1%, 'doctor-diagnosed asthma' 13.9%, 'ever experienced allergic rhinitis (AR) symptoms' 34.9%, 'AR-conjunctivitis symptoms in past 12 months' 10.7%, 'ever had hay fever' 7.8%, 'doctor-diagnosed hay fever' 11.2%, 'ever had eczema' 3.3% and 'doctor-diagnosed eczema' 3.2%. Kappa statistics demonstrated high within symptom category consistency for 'ever had asthma/doctor-diagnosed asthma' (0.728) and 'ever had eczema/doctor-diagnosed eczema' (0.906). Age and gender adjusted odds ratios (OR) were also consistently significant across wheeze and allergic rhinitis symptom categories [highest OR = 10.10 (95% CI 6.23-16.35) between allergic rhinoconjunctivitis and wheeze in past 12 months]. There is a high prevalence of ISAAC-based symptoms in school children in Ha Noi, Vietnam, often above global averages. The high level of association between atopic symptoms suggests some degree of reliability and validity. Childhood atopy symptom prevalence in Vietnam is more similar to that in developed countries rather than developing countries. PMID- 12911505 TI - Longitudinal follow-up of the changing gender ratio in asthma from childhood to adulthood: role of delayed manifestation in girls. AB - Boys suffer more often from asthma than girls, while in adults the gender ratio is reversed. It is not clear when exactly this change occurs and by what mechanism. From a cohort of all 5030 German 4th grade pupils (age 10 years) in Munich, 274 children with current asthma were identified (164 males, 110 females) through a questionnaire, and skin prick tests were performed. These subjects were re-evaluated at ages 14 and 20 years with a questionnaire. A random sample (n = 1000) of all 3538 German children without current asthma at age 10 was also re evaluated at age 20 (controls). At age 20, only 24.5% (21 males, eight females) of the initial asthma group still had symptoms, and their gender ratio remained male dominated. In the controls, 4.8% (48/1000) had current asthma at age 20 and these were predominantly female: 6.4% (31/485) of control girls vs. 3.3% (17/515) of boys (p = 0.022). Half of the new asthma cases had had no symptoms or diagnoses until age 10, and atopy at age 10 was not associated with subsequent asthma in these. Asthma at age 10 has no better prognosis in boys than in girls, and the mechanism of the changing gender ratio appears to be late incidence of asthma among girls. Because the latter constitute a considerable part of adult asthma cases, it appears important to further explore this asthma phenotype and the risk factors associated with it. PMID- 12911506 TI - Both allergen-specific CD4 and CD8 Type 2 T cells decreased in asthmatic children with immunotherapy. AB - Allergen-specific immunotherapy (IT) has been used for the treatment of atopic diseases since the turn of this century. The precise working mechanisms, however, remain to be clarified. The aim of this study was to investigate the role of particular subsets of allergen-specific T cells in the non-atopic individuals, untreated asthmatic children and the asthmatic children receiving immunotherapy. We collected peripheral blood from 16 untreated asthmatic children and 17 asthmatic children receiving immunotherapy over one and half years. All the patients were sensitive to mite allergen. Peripheral blood mononuclear cells (PBMC) were isolated and, in vitro, stimulated with crude mite extract to enrich the mite-specific T-cell population. After 14 days, the enriched mite-specific T cells were stimulated with phorbol-12-myristate-13-acetate (PMA) and ionomycin for intracellular detection of cytokines such as IFN-gamma, IL-4 in CD4+ and CD8+ T lymphocytes. The data here demonstrated that the levels of mite-specific IgG4 and IgA increased significantly in asthmatic children after immunotherapy. In addition, both IL-4 expressing CD4+ and CD8+ T cells were significantly lower in asthmatic children after immunotherapy compared with those of before treatment and the normal control (p < 0.05). In contrast, the frequency of IFN-gamma expressing CD4+ and CD8+ T cells did not significantly differ between untreated and SIT-treated groups. All these data suggested that decreased Type 2 CD4+ and CD8+ T cells might be closely correlated with the regulatory mechanisms of immunotherapy. PMID- 12911507 TI - Predictive features for persistence of atopic dermatitis in children. AB - Allergen exposure plays an important role in atopic dermatitis (AD). Because immunological mechanisms underlying asthma and AD have great similarities, we evaluated whether features such as allergen sensitization, immune response, disease severity and duration or allergen exposure could be considered predictive for AD persistence. Seventy-one AD children (age range 14-158 months) were enrolled and followed for 3 consecutive years for AD severity using the SCORAD index (SI). At enrollment, reactivity to inhalant and food allergens using the skin prick test (SPT) and house dust mite (HDM) atopy patch test (APT), and HDM allergens in house dust were evaluated. After 3 years, 38 children outgrew their AD (AD- group), while in 33 AD persisted (AD+ group). At enrollment, AD+ children had a higher SI, higher rate of positivity to SPT and APT for mites (p = 0.001), and higher environmental exposure to HDM allergens (p = 0.035). The AD+ children developed more respiratory symptoms in comparison to AD- children (p < 0.001). None of the AD- children presented APT positivity. In our study population, positivity of SPT and APT for HDM, environmental allergen exposure levels and severity of the disease at enrollment presented a significant predictive power towards AD persistence. Subjects with positive skin reactivity to HDM should be considered at risk of AD persistence and of possible development of allergic respiratory disorders. PMID- 12911508 TI - Serum concentration of macrophage-derived chemokine may be a useful inflammatory marker for assessing severity of atopic dermatitis in infants and young children. AB - Chemokines are responsible for the trafficking of leukocytes to sites of inflammation. Serum chemokine levels were previously shown to be increased in adult patients with atopic dermatitis (AD). We tested whether serum concentrations of chemokines, including macrophage-derived chemokine (MDC), thymus and activation-regulated chemokine (TARC), eotaxin (EOX), interferon gamma inducible protein 10 (IP-10) and monocyte chemotactic protein 1 (MCP-1), are useful inflammatory markers for assessing AD severity in infants and young children. To investigate this, we assessed the severity of AD clinically using the SCORing Atopic Dermatitis (SCORAD) index system. Serum chemokine concentrations were determined by sandwich enzyme immunoassay. Twenty AD patients with a median age of 2.1 years [interquartile range (IQR): 0.6-4.2] were recruited. Their SCORAD score was 23.5 (12.5-33.5). Serum concentrations of MDC, TARC, EOX, IP-10 and MCP-1 were 2551 (1978-3935), 1469 (1125-3070), 68 (57-85), 126 (101-226) and 518 (419-614) pg/ml, respectively. Serum MDC levels correlated with SCORAD (r = 0.608, p = 0.004) and its extent (r = 0.629, p = 0.003) and intensity (r = 0.557, p = 0.011) components. Serum TARC concentration showed weaker correlation with extent (r = 0.474, p = 0.035) and intensity (r = 0.465, p = 0.039) of skin involvement but not SCORAD. The median serum levels of MDC (3131 vs. 2394 pg/ml; p = 0.031) and EOX (80 vs. 61 pg/ml; p = 0.046) were also higher in children with moderate as compared with mild AD. The other chemokines did not correlate with AD severity. In conclusion, our results suggest that serum MDC concentration may be a useful inflammatory marker for assessing AD severity in infants and young children. PMID- 12911509 TI - Soluble E-selectin and soluble ICAM-1 levels as markers of the activity of atopic dermatitis in children. AB - The expression of adhesion molecules is up-regulated in the skin of atopic dermatitis (AD) patients, and the levels of the soluble adhesion molecules sE selectin and sICAM-1 have been reported to reflect the endothelial activation in the skin of AD patients. The objective of the study was to investigate the relationship between symptom score and levels of sE-selectin, sICAM-1 and sVCAM-1 before and after 2 weeks of treatment. Eighteen children with an exacerbation of AD were admitted and treated with corticosteroid dilutions under occlusive wet dressings (wet-wrap treatment). Symptom score (objective SCORAD) and levels of sE selectin, sICAM-1, and sVCAM-1 were assessed before and after 2 weeks of treatment. A significant correlation between the objective SCORAD before treatment and the level of sE-selectin (p < 0.05), but not the level of sICAM-1 (p = 0.7) or sVCAM-1 (p = 0.5) was observed. The treatment resulted in a high degree of remission, which was reflected by a significant decrease in the level of sICAM-1 (p < 0.01), whereas there was only a trend in the level of sE-selectin to decrease (p = 0.08). The level of sE-selectin after 2 weeks of treatment still correlated significantly with the objective SCORAD before treatment (p < 0.005). Soluble E-selectin is a relative objective marker for the severity of AD. SCORAD is a treatment-sensitive symptom of AD, whereas E-selectin may be a more stable underlying systemic representation of AD. PMID- 12911510 TI - Is maternal age at delivery related to childhood food allergy? AB - The prevalence of food allergies is increasing. Concurrently, the average maternal age at birth is also increasing. We conducted a preliminary study to evaluate whether maternal age at the time of delivery is associated with a food allergy in children. Case and control patients were identified among consecutive patients seen by one of us (AD) in the Allergy/Immunology program at the Children's Hospital Boston between 11/1/98 and 2/28/00. Case patients were born in Massachusetts and had evidence of clinical sensitivity and IgE to one or more food allergens (n = 58). Control patients were those born in Massachusetts who had a negative skin test and/or RAST to inhalant and/or food allergens (n = 96). A second comparison group consisted of all live births in Massachusetts in 1999 (n = 80,866). Information on maternal age at birth was missing from 3/58 (5%) of patients with a food allergy and 4/96 (4%) of the control patients, so these patients were not included in the analysis. The proportion of children whose mother was aged 30 and over at their birth was significantly higher in children with a food allergy than control patients (78% vs. 55% p = 0.005) and higher than all births in Massachusetts (78% vs. 53% p = 0.0002). Mothers of children with a food allergy had about three times greater odds of being aged 30 or over at the time of delivery than mothers in either of the comparison groups. Further exploration of the data using logistic regression showed that maternal age over 30 at delivery and being first born were independent predictors of the child having a food allergy. In this study, the presence of a food allergy in children was related to older maternal age at delivery. Additional studies are needed to further evaluate this relationship and its potential implication in preventive strategies for food allergies in children. PMID- 12911511 TI - The pattern of sesame sensitivity among infants and children. AB - Recently, we found sesame to be a major cause of severe IgE-mediated food allergic reactions among infants and young children in Israel. The purpose of this study was to describe the different patterns of sesame sensitivity. We have identified three subgroups among our patients (n = 32). Group I (n = 23, M/F; 14/9) consisted of cases with IgE-mediated sesame allergy. The mean age of the first allergic reaction was 11.7 months. Although the main clinical manifestation was urticaria/angiedema (n = 14, 60%), anaphylaxis was the presenting symptom in seven (30%) patients; all of them were younger than 1 year. Sixteen (70%) were found to be allergic to other foods, and other atopic diseases were identified in 18 (78%) patients. Three patients 'outgrew' their allergy within 1-2 years. Group II (n = 2) included cases in whom sesame allergy was ruled out based on a negative skin prick test (SPT) together with a negative open oral challenge. Group III (n = 7) consisted of patients that were found to be SPT positive for sesame as part of a screening for other food allergies. Although sesame products have become fashionable in westernized countries, early exposure may cause sesame to share eventually the same 'noteriety and fate' as peanut - a major cause of severe food allergic reactions. PMID- 12911512 TI - Anaphylaxis induced by pine nuts in two young girls. AB - Pine nuts are the seeds of Pinus pinea. There are few reported cases of allergy to pine nut. We describe two young girls with anaphylaxis caused by small amounts of pine nuts. Specific IgE to pine nut was demonstrated by skin prick tests and RAST but no IgE to other nuts and pine pollen was detected. The patients had IgE against a pine nut protein band with apparent molecular weights of approximately 17 kDa that could be considered as the main allergen. Our patients were monosensitized to pine nut and the 17-kDa protein could be correlated with the severe clinical symptoms. PMID- 12911514 TI - Food protein-induced enterocolitis syndrome--not only due to cow's milk and soy. AB - Over a of 7-year period, six patients (four males, two females aged 3-12 months) were diagnosed with food protein-induced enterocolitis syndrome (FPIES) triggered by foods other than cow's milk and soy: chicken in four, turkey in two, peas in one, and lentils in one (five patients reacted to more than one food type). All reactions developed within 2 h of ingestion of the allergenic food. To exclude other conditions with similar clinical symptoms, three infants underwent work-up for sepsis, one infant underwent work-up to exclude metabolic defects, and one underwent a barium enema to rule out intussusception. All were negative. Pediatricians should be aware that FPIES may be caused by foods other than cow's milk and soy, mainly chicken, turkey and foods from the legume family, and that it may present also in infants older than 6 months. PMID- 12911513 TI - Plasma cytokine profiles in patients with celiac disease and selective IgA deficiency. AB - Celiac disease (CD) and selective IgA deficiency (IgAD) are frequently associated, and share the same genetic background. The aim of the present study was to evaluate both Type 1 and 2 plasma cytokine levels in CD and in CD-IgAD. IL 2, TNF-alpha, IL-10, IL-4 and IL-13 plasma levels were measured both at diagnosis and after a gluten-free diet (GFD) in 32 CD patients, in 27 CD-IgAD patients and in 30 healthy controls. IFN-gamma levels were significantly higher in CD and CD IgAD than in controls, TNF-alpha displayed significantly higher levels in CD-IgAD when compared both with controls and with CD, and IL-2 was in CD-IgAD significantly increased respect to controls. Kinetics of the Type 1 cytokine plasma levels did not show a clear relationship with the GFD in both groups of CD patients, and particularly in those with IgAD. IL-4 and IL-13, both at diagnosis and after a GFD, were not significantly different in controls and in celiac patients (with and without IgAD). IL-10, whose production is stimulated by the TNF-alpha, had significantly higher plasma levels in CD-IgAD, but not in CD patients, with a significant decrease after a GFD. CD and especially CD-IgAD patients display persistently higher pro-inflammatory cytokine levels, suggesting a persistent state of activation of pro-phlogistic signals in CD, particularly when IgAD coexists. Serial measurement of serum IL-10 may be an adjunctive evaluating criterion in the follow-up of CD-IgAD patients. PMID- 12911515 TI - Unusual and severe disease course in a child with ataxia-telangiectasia. AB - Ataxia-telangiectasia (AT) is an autosomal recessive syndrome of combined immunodeficiency. Hallmarks of the disease comprise progressive cerebellar ataxia, oculocutaneous telangiectasia, cancer susceptibility and variable humoral and cellular immunodeficiency. We describe a patient with AT presenting with autoimmune haemolytic anaemia, neutropenia, hepatosplenomegaly, lymphadenopathy and hyper-IgM at the age of 6 months. At the age of 26 months she developed persistent fever, progressive lymphadenopathy and pulmonary nodular infiltrates, which were responsive to steroid therapy. PMID- 12911516 TI - Development of common variable immunodeficiency in a patient with Evans syndrome treated by autologous stem cell transplantation. AB - We describe a case report of a patient who developed common variable immunodeficiency (CVID) after autologous haematopoietic stem cell transplantation (SCT) for recurrent Evans syndrome. The disease manifested as attacks of haemolytic anaemia, thrombocytopenia and neutropenia from the age of 12 years. Presence of autoantibodies to blood elements was confirmed together with C4 deficiency. The patient also suffered from dermatitis herpetiformis Duhring without signs of coeliac disease. Autologous T cell-depleted peripheral blood stem cell (PBSC) transplant following conditioning regimen was performed at the age of 20 years. Immunological reconstitution was incomplete and 2 years after SCT he fulfilled laboratory criteria for common variable immunodeficiency (CVID). The patient was found to be a carrier of a risk haplotype for development of CVID DRB1*03/DQB1*0201. We conclude that T cell-depleted SCT here performed for autoimmune manifestations can hasten development of CVID in genetically predisposed patients. PMID- 12911517 TI - Response to von Bernuth et al.'s case report. PMID- 12911518 TI - The lack of suppressor of cytokine signalling-1 (SOCS1) protects mice from the development of cerebral malaria caused by Plasmodium berghei ANKA. AB - Cerebral malaria is a severe complication of infection with Plasmodium berghei ANKA involving the Th1 cytokines TNF-alpha and IFN-gamma. Suppressor of cytokine signalling-1 (SOCS1) is an important component in the regulatory cascade controlling inflammatory responses and signalling through IFN-gamma. Contrary to the expectation that SOCS1-deficient mice, in which IFN-gamma responses are uncontrolled and which are more sensitive to IFN-gamma, may show heightened susceptibility, mice lacking SOCS1 were protected from cerebral malaria. Unlike the controls and despite similar parasitaemia, infected SOCS1 null mice showed no inflammation or haemorrhaging in the brains. Mice lacking SOCS1 exhibited decreased splenic cellularity and a reduced ratio of CD4 : CD8 lymphocytes, which were maintained during infection. However, the ratio of IFN-gamma to IL-4 mRNA expression during infection was similar in SOCS1 -/- and control mice suggesting that a dramatic shift in the ratio of Th1 : Th2 responses does not account for the resistance to disease. Resistance conferred by the lack of SOCS1 is specific since the related SOCS2, also implicated in Th1-mediated responses, did not seem to be involved in the development of disease. Understanding the mechanism by which SOCS1 deficiency protects mice from cerebral malaria may allow the manipulation of its activity and alleviate pathology. PMID- 12911519 TI - Molecular and immunological characterization of the C-terminal region of a new Echinococcus granulosus Heat Shock Protein 70. AB - By screening an expression library of Echinococcus granulosus with IgE from sera of patients with cystic echinococcosis (CE) and allergic reactions, we isolated the C-terminal region of a new heat shock protein (HSP)70 of E. granulosus. The protein, named Eg2HSP70, has close homology with the C-terminal region of Dermatophagoides farinae and human HSP70. We investigated the humoral and cell mediated immune responses to this antigen in patients with CE grouped according to the presence of allergic reactions. Immunoblotting detected total IgG, IgG4 and IgE specific to Eg2HSP70 (83% of sera contained IgG, 31% IgG4 and 57% IgE). No significant difference was found in immunoglobulin percentages according to the presence of allergic reactions. Immunoblotting inhibition showed that no IgG or IgE specific to Eg2HSP70 cross-reacted with D. farinae or human HSP70. Eg2HSP70-stimulated PBMC from 26 patients produced significantly greater amounts of TNF-alpha, IFN-gamma, and IL-10 than unstimulated cultures in all patients, irrespective of the presence of allergic reactions (P < 0.05). They also produced significantly greater amounts of IL-4 than unstimulated cultures exclusively in patients with allergic reactions (P < 0.05). These findings show that Eg2HSP70 is a new antigenic molecule inducing both B and T cell responses. PMID- 12911520 TI - Immunomodulatory effects of Maxadilan and Phlebotomus papatasi sand fly salivary gland lysates on human primary in vitro immune responses. AB - Leishmaniasis is a parasitic disease transmitted by the bite of Leishmania infected sand flies. Here we show for the first time the ability of Maxadilan (Max), a vasodilatory peptide isolated from the sand fly Lutzomyia longipalpis, and salivary gland lysate (SGL) from Phlebotomus papatasi to decrease the secretion of Type 1 cytokines and to enhance the production of the Type 2 cytokine interleukin (IL)-6 by human peripheral blood mononuclear cells (PBMC) and monocytes. We found Max decreased the secretion of interferon (IFN)-gamma and IL-12p40 by PBMC and TNF-alpha by monocytes. SGL reduced the production of IFN gamma by PBMC. In contrast, production of the Type 2 cytokine IL-6 was increased in Max or SGL-exposed cells. Finally, we determined that Max interacts with human cells through at least the pituitary adenylate cyclase activating polypeptide receptor. These results show that sand fly salivary gland components have an immunomodulatory effect on human cells, and this has important implications for the development of vaccines against leishmaniasis for humans. PMID- 12911521 TI - Vaccination against Fasciola hepatica infection using a Schistosoma mansoni defined recombinant antigen, Sm14. AB - Fasciola hepatica is the causative agent of fasciolosis in many areas in America, Europe, Africa, Asia and Australia. There is an urgent need for improved methods to control the parasite's transmission. We describe the use of an experimental vaccine based on a recombinant antigen cloned from another parasite, Schistosoma mansoni (Sm14), that induces high levels of cross protection in mice against both S. mansoni and F. hepatica. Sheep and mice vaccinated with Sm14 were significantly protected against challenge infection with metacercariae of Fasciola hepatica and were completely free of the histopathological hepatic damage related to liver fluke infection. The vaccine will provide a valuable new tool to aid in transmission control of this economically important disease. PMID- 12911522 TI - The site of cutaneous infection influences the immunological response and clinical outcome of hamsters infected with Leishmania panamensis. AB - We determined that the site of inoculation (foot or snout) influences the clinical evolution and immune responses of hamsters infected with Leishmania (Viannia) panamensis. Hamsters infected in the snout showed (i) a more rapid and severe lesion evolution at multiple time points (P < 0.05), (ii) a more extensive inflammatory infiltrate and tissue necrosis, (iii) a higher tissue parasite burden, (iv) a higher antibody titre (P < 0.01), but lower antigen-specific spleen cell proliferative response (P = 0.02), and (v) a slower response to anti leishmanial drug treatment (P < 0.002). In both inoculation groups there was co expression of type 1 (IFN-gamma and IL-12) and some type 2 (IL-10 and TGF-beta, but not IL-4) cytokines in the cutaneous lesions and spleen. Early in the course of infection, hamsters infected in the snout showed higher expression of splenic IL-10 (P = 0.04) and intra-lesional IFN-gamma (P = 0.02) than foot infections. No expression of IL-12p40 or IL-4 was detected. During the chronic phase, snout lesions expressed more IFN-gamma (P = 0.001), IL-12p40 (P = 0.01), IL-10 (P = 0.009) and TGF-beta (P = 0.001), and the level of expression of each of these cytokines correlated with lesion size (P < or = 0.01). These results suggest that the site of infection influences the clinical outcome in experimental cutaneous leishmaniasis, and that the expression of macrophage-deactivating type 2 cytokines and/or an exaggerated type 1 proinflammatory cytokine response may contribute to lesion severity. PMID- 12911523 TI - Pharmacological assessment of the role of nitric oxide in mice infected with lethal and nonlethal species of malaria. AB - This pharmacological investigation sought to determine whether nitric oxide (NO) had an antiparasitic effect and/or mediated pathology in mice infected with nonlethal P. chabaudi or lethal P. berghei. Nitric oxide synthase (NOS) inhibitors were evaluated for their ability to inhibit the rise in reactive nitrogen intermediates (RNI) induced by bacterial lipopolysaccharide (LPS) in mice. The more effective compound, aminoguanidine (AG) inhibited the rise in RNI induced by P. chabaudi and increased mortality, but had no effect on parasitaemia. Inducers and donors of NO were screened for their ability to increase RNI and the most effective agents evaluated for their ability to modify P. berghei infection. S-Nitrosoglutathione had little effect, but LPS decreased parasitaemia and mortality. An inconsistent relationship is evident between the abilities of these agents to modify NO activity and their effects on malaria in mice. Increased mortality in mice with P. chabaudi treated with AG indicates a reduction in resistance. The absence of an effect on parasitaemia by a NOS inhibitor or NO donor indicates either RNI have insignificant antimalarial action in vivo or the efficacy of the compounds is inadequade. Resistance to P. berghei in LPS-treated mice demonstrates an antiparasitic effect, but this may be attributable to factors other than NO. PMID- 12911524 TI - Immunoglobulin profiles in a murine intermediate host model of resistance for Echinococcus granulosus infection. AB - We have shown previously that primary infection of Chinese Kunming (CKM) mice with Echinococcus granulosus oncospheres is protective against subsequent challenge. Nine groups of mice were infected with the oncospheres of E. granulosus by different routes (intraperitoneal, subcutaneous or intravenous injection). After infection, serum was collected after different periods of time and serum antibodies were tested by ELISA against oncospheral proteins and hydatid cyst fluid antigens. The results indicated that CKM mice produced low levels of antibodies before a secondary challenge infection given 3 weeks later by a different route. Most mice did not evoke significant antibody responses against oncospheral antigens until 5 weeks after infection. The level of IgG, especially IgG1 against oncospheral antigens increased from week 4 post-infection (p.i.), to a maximum at week 9 p.i. In addition, antibodies against hydatid cyst fluid antigens increased at the same time as the recognition of oncospheral antigens. Immunoblots using hydatid cyst fluid showed that the first antigen that was recognized - an 8-kDa protein, possibly the smallest subunit of Antigen B - appeared 5-6 weeks p.i. and reactivity to this molecule was intensive at week 9 p.i. The results suggest that protection against secondary infection was not principally antibody-mediated during the initial phases of infection, when cellular immune responses may play a pivotal role in the protective mechanism. PMID- 12911525 TI - Kinetics of eosinophil and IgE-mast cell changes following infection with Angiostrongylus costaricensis in Wistar rats. AB - Human abdominal angiostrongyliasis is a severe eosinophilic disease caused by Angiostrongylus costaricensis. Previous studies have demonstrated that wild rodents are critically involved as definitive hosts to this nematode in nature. In this study, we have evaluated the susceptibility of Wistar rats (Rattus norvegicus) to A. costaricensis infection. Kinetics of parasitological and pathological changes, including the number of adult worms recovered from mesenteric arteries, and of IgE, mast cell and eosinophil levels in several compartments have been assessed. The oral inoculation of third-stage larvae (L3) into adult Wistar rats led to a marked accumulation of worms in the branches of the mesenteric arteries 25 and 50 days post-inoculation. Intense bone marrow eosinophilia ranging from 7 to 50 days was accompanied by marked accumulation of eosinophils in the blood, peritoneal and bronchoalveolar spaces. Eosinophilic periarteritis, oedema and granuloma in the intestinal and lung tissues were also histologically evident. Total serum IgE and specific anti-parasite IgE peaked at 25 days post-infection, as measured by ELISA and by the passive cutaneous anaphylaxis test, respectively. At that time point, there was a drastic reduction in the number of intact mast cells in the peritoneal effluent. These findings indicate that Wistar rats are permissive to A. costaricensis infection. IgE-mast cell activation and massive tissue eosinophil infiltration are marked features in the process and are likely to play a crucial role in the immune-response evoked by this parasite. PMID- 12911526 TI - Stewart and beyond: new models of acid-base balance. AB - The Henderson-Hasselbalch equation and the base excess have been used traditionally to describe the acid-base balance of the blood. In 1981, Stewart proposed a new model of acid-base balance based upon three variables, the "strong ion difference" (SID), the total weak acids (ATot), and the partial pressure of carbon dioxide (Pco2). Over 20 years later, Stewart's physiochemical model still remains largely unknown. In this review, we will present both the traditional and the Stewart models of acid-base balance and then derive each using an "ion equilibrium method." Modern theories of acid-base balance may be useful toward the understanding of complex acid-base disorders. PMID- 12911527 TI - Refinement of the critical region for MCKD1 by detection of transcontinental haplotype sharing. AB - BACKGROUND: Autosomal-dominant medullary cystic kidney disease type 1 (MCKD1) [OMIM 174000] is a hereditary nephropathy that leads to renal salt wasting and end-stage renal failure at a median age of 62 years. In a Welsh MCKD1 kindred we have recently demonstrated linkage to the MCKD1 locus on chromosome 1q23.1 and refined the critical MCKD1 region to <3.3 Mb. METHODS: In order to refine the candidate gene region for MCKD1, high-resolution haplotype analysis in three large kindreds with MCKD1 was performed. RESULTS: We report here on high resolution haplotype analysis in this Welsh kindred, as well as in the Arizona kindred, which was used for the first definition of MCKD as a disease entity, and in a kindred from the Dutch/German border. We detected extensive haplotype sharing among all affected individuals of all three kindreds. Scrutinization of the genealogy of the Arizona kindred revealed an origin from Germany in the 17th century, thereby providing historical data for haplotype sharing by descent at the MCKD1 locus. CONCLUSION: Under the hypothesis of haplotype sharing by descent, we refined the critical genetic interval to <650 kb, thus enabling candidate gene analysis. PMID- 12911528 TI - Enlarged nephrons and severe nondiabetic nephropathy in hepatocyte nuclear factor 1beta (HNF-1beta) mutation carriers. AB - BACKGROUND: Mutations in hepatocyte nuclear factor-1beta (HNF-1beta) lead to a syndrome with diabetes and urogenital malformations [maturity onset of diabetes of the young, type 5 (MODY5)]. The aim of this study was to perform a clinicopathologic investigation of the renal disease in members of a Norwegian family with the HNF-1beta mutation R137-K161del. METHODS: The study was based on long-term clinical observations of five mutation carriers, combined with renal biopsies from four of these. The biopsies were examined by light microscopy, immunohistochemistry, and transmission electron microscopy. The diameter of the glomerulus, proximal and distal tubules, in addition to thickness of the glomerular basement membrane (GBM), were measured in light microscopic slides and transmission electron micrographs. The results were compared with biopsies from adult patients with diabetic glomerulopathy, glomerulonephritis, and/or benign nephrosclerosis, and children with minimal-change glomerulopathy or glomerulonephritis, respectively. RESULTS: Clinically, there was a wide intrafamilial variation from stable or slightly increasing serum creatinine to progressive renal failure and end-stage renal disease (ESRD). In all cases, the kidney disease was diagnosed prior to diabetes. Hypertrophy of the proximal and distal tubules as well as enlarged glomeruli were found in three of four mutation carriers. Essentially normal nephrons were found in the 10-year-old boy. The thickness of the GBM was considered near normal in all mutation carriers. Oligomeganephronia was found in one patient. CONCLUSION: Histopathologic and morphometric studies of kidney biopsies from four carriers of an HNF-1beta mutation revealed enlarged glomeruli and tubular structures. Long-term clinical follow-up demonstrated that the renal disease developed prior to and independently of diabetes. Finally, there is a wide phenotypic variation of the renal disease caused by HNF-1beta mutations. PMID- 12911529 TI - Fabry disease: detection of undiagnosed hemodialysis patients and identification of a "renal variant" phenotype. AB - BACKGROUND: Fabry disease is an X-linked recessive lysosomal storage disease resulting from deficient alpha-galactosidase A (alpha-Gal A) activity. Renal failure is a major debilitating complication in classically affected males. To determine if this disorder is underdiagnosed in patients with end-stage renal disease (ESRD), the frequency of unrecognized males with Fabry disease on chronic hemodialysis was determined. METHODS: Plasma alpha-Gal A activity was measured in 514 consecutive males with ESRD on hemodialysis. Patients with low alpha-Gal A activity were evaluated clinically and their alpha-Gal A mutations were determined. RESULTS: Six (1.2%) of 514 hemodialysis patients had low plasma alpha Gal A activities and a previously identified (E66Q, A97V, M296I) or novel (G373D) missense mutation. At ages 30 to 68 years, five patients lacked the classic manifestations of angiokeratoma, acroparesthesias, hypohidrosis, and ocular opacities, while the sixth lacked angiokeratoma and ocular changes. Five had left ventricular hypertrophy (LVH). CONCLUSION: The clinical spectrum of Fabry disease includes a "renal variant" phenotype in patients without classic symptoms who develop ESRD. Affected males undergoing hemodialysis or renal transplantation can be readily diagnosed by plasma alpha-Gal A assays. These patients and their family members may benefit from enzyme replacement therapy for the later, life threatening cardiovascular and cerebrovascular complications of Fabry disease. PMID- 12911530 TI - Analysis of renal tubular electrolyte transporter genes in seven patients with hypokalemic metabolic alkalosis. AB - BACKGROUND: Disorders that manifest hypokalemic metabolic alkalosis, such as Bartter's syndrome and Gitelman's syndrome, are caused by the malfunction of renal tubular electrolyte transporters. Bartter's syndrome may be linked to dysfunction of Na-K-2Cl cotransporter (NKCC2), renal outer medullary K channel (ROMK), or Cl channel Kb (ClC-Kb), while Gitelman's syndrome may be linked to Na Cl cotransporter (NCCT) dysfunction. However, previous genetic analyses in these syndromes have included many heterozygotes for each gene and there has been no further analysis of other genes. Thus, to clarify the interaction of these transporter genes, in the present study we investigated all 4 transporter genes in 7 patients with hypokalemic metabolic alkalosis. METHODS: Seven patients from 5 families (patients A-G) were collected, and a mutation analysis of the 4 renal electrolyte transporter genes was performed by direct sequencing. RESULTS: We identified 12 mutations in these 7 patients. Three mutations (del245Y in NKCC2, R1009X in NCCT, V524I in ClC-Kb) have not been reported previously. In NKCC2 gene screening, patient A was homozygous for del245Y. In ClC-Kb gene screening, L27R was detected in patients B, D, and E. V524I was detected in patient C. Both T562M and E578K were observed in patients B and E. In NCCT gene screening, patients B-G shared a common novel mutation, R1009X, and patients D, E, F, and G carried this mutation in both alleles. Patients B and C carried R1009X in one allele, and a 6 amino acid insertion in exon 6 and L849H in another allele, respectively. The 4 other mutations did not result in any amino acid exchange. Despite the NCCT gene mutation, patients C and E showed normomagnesemia. CONCLUSION: Our findings demonstrate that in Bartter's and Gitelman's syndromes, it may not be uncommon to see mutations in several causative transporter genes. PMID- 12911532 TI - Prenatal exposure to nicotine modifies kidney weight and blood pressure in genetically susceptible rats: a case of gene-environment interaction. AB - BACKGROUND: Epidemiologic studies suggest that in utero exposure to maternal smoking is associated with elevated blood pressure (BP) later in life. Our aims were: (1) to examine effects of intrauterine exposure to nicotine on BP and hypertension target-organ size in rats; and (2) to investigate whether such effects depend on genetic background, by studying two genetically distinct strains of rats: the spontaneously hypertensive rat (SHR) and the normotensive Brown Norway (BN) rat. METHODS: Nicotine or saline was administered to dams via subcutaneous osmotic minipumps throughout gestation. In nine-week-old male offspring, we measured BP and heart rate, assessed the weight of kidneys and heart, and determined fasting levels of glucose, insulin, triglycerides (TG), and cholesterol. We also measured gene expression of the insulin-like growth factor (IGF) system in the liver and kidneys. RESULTS: SHR and BN offspring differed in their response to intrauterine exposure to nicotine. SHR exposed to nicotine (vs. saline) exhibited higher BP (P < 0.02) and serum cholesterol levels (P = 0.01), and lower kidney weight (P < 0.0001). In contrast, BN rats did not demonstrate differences between the nicotine and saline groups in these variables, but the nicotine-exposed BN rats showed a significant up-regulation in the gene expression of IGF-1 in the liver (P < 0.0001) and IGF receptor in the kidney (P = 0.006). CONCLUSION: These results suggest that intrauterine exposure to nicotine alters the cardiovascular system depending on the genetic background and, as such, supports the notion that the intrauterine environment interacts with genes in determining an individual's health later in life. PMID- 12911531 TI - Lipoproteins in the DCCT/EDIC cohort: associations with diabetic nephropathy. AB - BACKGROUND: Lipoproteins may contribute to diabetic nephropathy. Nuclear magnetic resonance (NMR) can quantify subclasses and mean particle size of very low density lipoprotein (VLDL), low density lipoprotein (LDL), and high density lipoprotein (HDL), and LDL particle concentration. The relationship between detailed lipoprotein analyses and diabetic nephropathy is of interest. METHODS: In a cross-sectional study, lipoproteins from 428 women and 540 men from the Diabetes Control and Complications Trial/Epidemiology of Diabetes Interventions and Complications (DCCT/EDIC) cohort were characterized by conventional lipid enzymology, NMR, apolipoprotein levels, and LDL oxidizibility. Linear regression was performed for each lipoprotein parameter versus log albumin excretion rate (AER), with and without covariates for age, diabetes duration, HbA1c, hypertension, body mass index, waist-hip ratio, and DCCT treatment group. Significance was taken at P < 0.05. RESULTS: By multivariate analysis, conventional profile, total triglycerides, total- and LDL cholesterol, but not HDL cholesterol, were associated with AER. NMR-determined large, medium, and small VLDL were associated with AER in both genders (except large VLDL in women), and intermediate density lipoprotein (IDL) was associated with AER (men only). LDL particle concentration and ApoB were positively associated with AER (in men and in the total cohort), and there was a borderline inverse association between LDL diameter and AER in men. Small HDL was positively associated with AER and a borderline negative association was found for large HDL. No associations were found with ApoA1, Lp(a), or LDL oxidizibility. CONCLUSION: Potentially atherogenic lipoprotein profiles are associated with renal dysfunction in type 1 diabetes and further details are gained from NMR analysis. Longitudinal studies are needed to determine if dyslipoproteinemia can predict patients at risk of nephropathy, or if lipoprotein-related interventions retard nephropathy. PMID- 12911533 TI - Oxidative stress and kidney dysfunction due to ischemia/reperfusion in rat: attenuation by dehydroepiandrosterone. AB - BACKGROUND: The pathogenesis of ischemia/reperfusion (I/R) involves generation of reactive oxygen and nitrogen species. This in vivo study investigates the effect of dehydroepiandrosterone (DHEA), a physiologic steroid with antioxidant properties, on oxidative balance and renal dysfunctions induced by monolateral I/R. METHODS: Normal and DHEA-treated rats (4 mg/day x 21 days, orally) were subjected to monolateral renal I/R (30 minutes/6 hours). The oxidative state was determined by measuring hydrogen peroxide level and activities of glutathione peroxidase, catalase, and superoxide dismutase. Tumor necrosis factor-alpha (TNF alpha) and nitric oxide production and inducible nitric oxide synthase (iNOS) levels were also measured. Hydroxynonenal content was used to probe lipid peroxidation. Functional parameters determined were creatinine levels and Na/K ATPase activity. Immunohistochemical and morphologic studies were also performed. RESULTS: A markedly pro-oxidant state was evident in the kidney of rats subjected to I/R. Both hydrogen peroxide and reactive nitrogen species (nitric oxide and iNOS) increased, whereas antioxidants decreased. Oxidant species induce TNF-alpha increase, which, in turn, produces lipoperoxidative processes, as documented by the increased hydroxynonenal (HNE) level. As final result, impaired renal functionality, hydropic degeneration, and vacuolization of proximal convolute tubules were observed in kidneys of I/R rats. DHEA pretreatment improved the parameters considered. CONCLUSION: I/R induces oxidative stress and consequently damages the proximal convolute renal tubules. Rats supplemented with DHEA and subjected to I/R had reduced pro-oxidant state, oxidative damage, and improved renal functionality, indicating an attenuation of oxidative injury and dysfunctions mediated by I/R. PMID- 12911534 TI - TGF-beta isoforms in renal fibrogenesis. AB - BACKGROUND: Transforming growth factor-beta1 (TGF-beta1) is generally considered to be the major or predominant isoform involved in fibrosis, with the roles of TGF-beta2 and -beta3 being less clear. Because anti-TGF-beta-specific isoform treatment is in development, it is important to know more precisely about isoform action. Here we compared the actions of each isoform on production and degradation of extracellular matrix proteins by cultured rat mesangial cells, renal fibroblasts, and tubular epithelial cells. We investigated endogenous production of each isoform, the effect of adding one isoform on the production of the other isoforms, and the response to addition of isoform combinations on matrix protein production. Isoform-specific antibodies were used to determine the relative contribution of these isoforms to matrix protein production. METHODS: Each cell type was treated with TGF-beta (0.01 to 10 ng/mL) alone or in different combinations. Living cell number was determined by 3-[4,5]dimethylthiazol-2,5 diphenyltetrazolium bromide (MTT) assay. Supernatant fibronectin and TGF-beta isoform concentration were measured by enzyme-linked immunosorbent assay (ELISA). Collagen and proteoglycan production were measured by [3H]-proline and [35S] sulfate incorporation, respectively. Matrix protein and TGF-beta isoform gene expression were determined by Northern blot. Release of 3H from preformed radiolabeled matrix by fibroblasts was used as a measure of matrix degradation. RESULTS: Each isoform increased matrix protein synthesis and reduced matrix degradation by renal cells similarly. Combination of TGF-beta isoforms showed additive effects. No antifibrotic effect was observed with TGF-beta3. TGF-beta1 increased -beta2 and -beta3 production in a small and inconsistent manner. In contrast, TGF-beta2 and -beta3 stimulated TGF-beta1 in all three cell types. Eighty percent of TGF-beta3's fibrogenic effect was mediated by TGF-beta1. A pan specific antibody to TGF-beta most effectively blocked plasminogen activator inhibitor type 1 (PAI-1) synthesis by epithelial cells under oxidative stress. CONCLUSION: All three TGF-beta isoforms have fibrogenic effects on renal cells. TGF-beta2 and TGF-beta3 effects may be partially mediated by TGF-beta1. These data suggest that blockade of all isoforms together may yield the best therapeutic effect in reducing renal fibrosis. PMID- 12911535 TI - Combining lisinopril and l-arginine slows disease progression and reduces endothelin-1 in passive Heymann nephritis. AB - BACKGROUND: Despite angiotensin-converting enzyme (ACE) inhibition is a very powerful therapy, it may not be uniformly renoprotective in patients with proteinuric nephropathies who might refer late in the course of the disease. In accelerated passive Heymann nephritis (PHN), a severe rat model of human membranous nephropathy, with proteinuria and increased urinary excretion of endothelin-1 (ET-1), early treatment with an ACE inhibition limited proteinuria as well as the exuberant formation of renal ET-1, while late treatment reduced urinary proteins not to a significant extent. Since biologic effects and production of ET-1 within the kidney are counteracted by nitric oxide, we studied the effect of combining lisinopril and l-arginine, the natural precursor of nitric oxide, starting late in the disease. METHODS: Uninephrectomized PHN rats were divided in four groups (N = 10) and daily given orally: vehicle; 1.25 g/L l arginine; 40 mg/L lisinopril; and l-arginine + lisinopril. Treatments started at 2 months, when rats had massive proteinuria, until 9 months. Six normal rats served as control. RESULTS: Increase in systolic blood pressure was significantly limited by l-arginine. Lisinopril alone and the combination were more effective. Renal function impairment was not affected by l-arginine, partially ameliorated by ACE inhibitor and normalized by the combined therapy. In rats given l arginine, proteinuria levels were similar to vehicle. ACE inhibitor kept proteinuria at values comparable to pretreatment and numerically lower than vehicle. Addition of l-arginine to lisinopril was more effective, with values significantly lower than vehicle. Glomerular and tubular changes were limited by the ACE inhibitor and further ameliorated by the combined therapy. Exaggerated urinary ET-1 of PHN was reduced by 23% and 40% after l-arginine and lisinopril, respectively, and by 62% with the combination. Defective urinary excretion of cyclic guanosine monophosphate (cGMP) was partially restored by lisinopril, while normalized by the combined therapy. CONCLUSION: Combining l-arginine with ACE inhibitors would represent a novel strategy for patients with severe nephropathy not completely responsive to ACE inhibition. Restoring the nitric oxide/ET-1 balance could be of benefit in halting renal disease progression. PMID- 12911536 TI - Effect of immunosuppression on damage, leukocyte infiltration, and regeneration after severe warm ischemia/reperfusion renal injury. AB - BACKGROUND: Post-ischemia/reperfusion (I/R) damage, accompanied by leukocyte infiltration, is unavoidable in renal transplantation, as is the need for immunosuppressive treatment. Influence of immunosuppressive treatment on post-I/R renal damage, nonalloimmune cellular infiltration, and regeneration is not well studied. METHODS: Uninephrectomized inbred LEW rats were submitted to warm renal ischemia of 45 minutes/60 minutes, and received different immunosuppressive regimens: cyclosporine (CsA) 10 mg/kg/day subcutaneously in the neck daily, or mycophenolate mofetil (MMF) 20 mg/kg/day by daily oral gavage. Control animals underwent sham operation (unilateral nephrectomy) with immunosuppressive treatment or ischemia with vehicle administration. In addition the effect of MMF/mycophenolic acid (MPA) on renal tubule cell proliferation in culture was studied with bromodeoxyuridine incorporation. RESULTS: The post-I/R interstitial cellular infiltration/proliferation consisted mainly of mononuclear leukocytes [first monocytes/macrophages (Mo/MPhi) followed by CD4+ cells]. This mononuclear cell infiltration became apparent 24 hours after injury at the time of acute tubular necrosis, and was most prominent during the phase of regeneration. Severe I/R combined with CsA aggravated morphologic damage and dysfunction, without effect on tubular cell proliferation and tubular regeneration. Early leukocyte infiltration was qualitatively and quantitatively comparable to control animals, yet decreased moderately later in time. In contrast, MMF in combination with severe I/R did not influence initial morphologic damage and dysfunction. Although the initial leukocyte infiltration was comparable to control animals, the subsequent mononuclear cell accumulation, especially CD4 T cells decreased dramatically during MMF treatment. This was concomitant with a decrease of tubular cell proliferation and hence tubular regeneration. Increasing MPA concentrations in renal tubular cell culture caused a significant decrease in total cell number, and an almost arrest of bromodeoxyuridine incorporation, as measurement of cell proliferation. CONCLUSION: Immunosuppressive treatment with CsA or MMF affected significantly and in a different manner post-I/R renal morphologic damage, interstitial leukocyte, accumulation and regeneration. PMID- 12911537 TI - Cellular responses to hypoxia after renal segmental infarction. AB - BACKGROUND: Hypoxia is believed to play an important role in the pathogenesis of acute and chronic kidney disease. However, the impact of low oxygen tensions on cellular functions in the kidney and potential adaptive responses are poorly understood. METHODS: In order to assess the effects of regional hypoxia, we induced large segmental renal infarcts in rats by renal artery branch ligation to create an oxygen gradient vertical to the corticomedullary axis and studied the effects on cell morphology, the induction of hypoxia-inducible transcription factors (HIF), the expression of HIF target genes, and cell proliferation. RESULTS: Pimonidazol protein adduct immunohistochemistry, a marker for severe tissue hypoxia, verified a continuous area of hypoxic renal tissue extending from the cortex to the papilla, in which tubular necrosis developed subsequently. Within this area local sparing of pimonidazol staining and tissue preservation was found around arcuate veins, indicating regional oxygen supply via diffusion from venous blood. HIF-1alpha was up-regulated within 1 hour and for up to 7 days predominantly in the border zone of the infarct in tubular cells, glomerular cells, resident interstitial cells, capillary endothelial cells, and infiltrating macrophages. HIF-2alpha expression was less prominent and confined to resident and infiltrating peritubular cells in the cortex. HIF expression was colocalized with regional up-regulation of the hypoxia-inducible genes heme oxygenase-1 and vascular endothelial growth factor (VEGF), and was followed by capillary and tubular proliferation. CONCLUSION: Our findings illustrate a marked potential of renal tissue to respond to regional ischemia and initiate adaptive reactions, including angiogenesis. PMID- 12911538 TI - Antisense oligonucleotides against collagen-binding stress protein HSP47 suppress peritoneal fibrosis in rats. AB - BACKGROUND: Peritoneal fibrosis is a serious complication in patients on continuous ambulatory peritoneal dialysis (CAPD), but the molecular mechanism of this process remains unclear. Heat shock protein 47 (HSP47), a collagen-specific molecular chaperone, is essential for biosynthesis and secretion of collagen molecules, and is expressed in the tissue of human peritoneal fibrosis. In the present study, we examined the effect of HSP47 antisense oligonucleotides (ODNs) on the development of experimental peritoneal fibrosis induced by daily intraperitoneal injections of chlorhexidine gluconate (CG). METHODS: HSP47 antisense or sense ODNs were injected simultaneously with CG from day 14, after injections of CG alone. Peritoneal tissue was dissected out 28 days after CG injection. The expression patterns of HSP47, type I and type III collagen, alpha smooth muscle actin (alpha-SMA), as a marker of myofibroblasts, ED-1 (as a marker of macrophages), and factor VIII were examined by immunohistochemistry. RESULTS: In rats treated with CG alone, the submesothelial collagenous compact zone was thickened, where the expression levels of HSP47, type I and type III collagen and alpha-SMA were increased. Marked macrophage infiltration was also noted and the number of vessels positively stained for factor VIII increased in the CG-treated group. Treatment with antisense ODNs, but not sense ODNs, abrogated CG-induced changes in the expression of HSP47, type I and III collagen, alpha-SMA, and the number of infiltrating macrophages and vessels. CONCLUSION: Our results indicate the involvement of HSP47 in the progression of peritoneal fibrosis and that inhibition of HSP47 expression might merit further clinical investigation for the treatment of peritoneal fibrosis in CAPD patients. PMID- 12911539 TI - Nephritogenic autoantibodies but absence of nephritis in Il-12p35-deficient mice with pristane-induced lupus. AB - BACKGROUND: There is strong evidence that Th1 cytokines are essential for disease in murine models of lupus. Interleukin-12 (IL-12) is essential for Th1 cell differentiation and induces interferon-gamma (IFN-gamma) production. Paradoxically, it has been suggested that an IL-12 defect drives the pathogenesis of lupus, although its precise role remains unclear. We investigated the role of IL-12 for lupus-like disease induced by pristane. IL-12p35-deficient (-/-) and control (+/+) BALB/c mice were treated with pristane or phosphate-buffered saline (PBS). METHODS: Proteinuria was measured and renal pathology evaluated 10 months after treatment. Sera were analyzed for autoantibodies and total immunoglobulin levels. Cytokine expression and production was analyzed. RESULTS: Pristane induced nephritogenic autoantibodies and renal immunoglobulin and complement deposition in both IL-12 -/- and +/+ mice. However, proliferative pathology and proteinuria were absent in IL-12-/- mice, whereas pristane induced severe nephritis in one third of the +/+ mice. As expected, cytokine balance was skewed toward a Th2 response in pristane-treated IL-12 -/- mice. CONCLUSION: These data indicate that renal immune complex deposition can occur in the absence of IL 12p35, but that structural renal damage requires the presence of IL-12p35 or mediators induced by this molecule, such as IFN-gamma. In contrast to the abrogation of nephritogenic autoantibodies by the lack of IFN-gamma, such antibodies are induced by pristane in IL-12p35-deficient mice. Absence of structural renal disease, despite the presence of nephritogenic autoantibodies in pristane-treated IL-12-/- mice, indicates that antibody deposition alone is not sufficient for the development of lupus nephritis in this model. PMID- 12911540 TI - Mast cell infiltration and chemokine expression in progressive renal disease. AB - BACKGROUND: Mast cells are growth factor-rich, bone marrow-derived cells that infiltrate injured tissue where they have been implicated in the pathogenesis of progressive fibrosis. METHODS: Mast cell infiltration and the expression of related chemoattractants was examined following 5/6 nephrectomy, a model of progressive, nonimmune-mediated renal injury. In addition, expression of the profibrotic cytokine, transforming growth factor-beta (TGF-beta) within mast cells and the effects of renoprotective therapy with angiotensin-converting enzyme (ACE) inhibition were also determined. RESULTS: Renal injury was accompanied by mast cell infiltration, in close proximity to areas of tubulointerstitial fibrosis. Mast cells displayed toluidine blue metachromasia and were immunopositive for TGF-beta1 as well as chymase and tryptase. The expression of several mast cell chemokines, including stem cell factor, interleukin-8 (IL-8), and also TGF-beta1, were increased in 5/6 nephrectomized kidneys. ACE inhibition with ramipril led to a reduction in renal injury in association with attenuation of mast cell infiltration and chemokine expression. CONCLUSION: Mast cell infiltration and related chemokine expression are prominent and early features following renal mass reduction and may contribute pathogenetically to progressive renal injury. PMID- 12911541 TI - Rat glomerular epithelial cells produce and bear factor H on their surface that is up-regulated under complement attack. AB - BACKGROUND: Factor H is a potent complement inhibitory molecule that is primarily produced by the liver and appears in plasma as a soluble protein. Yet there is evidence that other cells, including those in the kidney, can produce factor H, and that it can be cell-associated as well as present as a plasma protein. Here we studied factor H in rat glomerular epithelial cells (GEC). METHODS: A polyclonal antibody to factor H was used to identify factor H protein. A polymerase chain reaction (PCR)-based strategy was utilized to clone the full length cDNA of GEC factor H. The relative quantity of factor H mRNA was measured by quantitative reverse transcription (RT)-PCR in cultured GEC exposed to complement activation and in the passive Heymann nephritis (PHN) model of membranous nephropathy. RESULTS: By immunofluorescence microscopy, factor H protein was present on the plasma membranes of cultured GEC. Based upon Western blot studies, this appeared to be the full-length 150 kD factor H protein. Factor H cDNA cloned from GEC was identical to the newly deposited sequence for rat liver factor H cDNA. In cultured GEC in which complement was activated, factor H mRNA increased over time. Similarly, in the PHN model in which complement was activated on GEC in vivo, factor H mRNA and protein also increased over time. CONCLUSION: Cultured GEC and glomeruli express factor H mRNA and protein. As modeled both in vitro and in vivo in the rat, factor H is up-regulated in membranous nephropathy. This is likely to be a direct response of GEC to complement attack and may represent a protective response of this cell. PMID- 12911542 TI - Classification and rescue of ROMK mutations underlying hyperprostaglandin E syndrome/antenatal Bartter syndrome. AB - BACKGROUND: Mutations in the renal K+ channel ROMK (Kir 1.1) cause hyperprostaglandin E syndrome/antenatal Bartter syndrome (HPS/aBS), a severe tubular disorder leading to renal salt and water wasting. Several studies confirmed the predominance of alterations of current properties in ROMK mutants. However, in most of these studies, analysis was restricted to nonmammalian cells and electrophysiologic methods. Therefore, for the majority of ROMK mutations, disturbances in protein trafficking remained unclear. The aim of the present study was the evaluation of different pathogenic mechanisms of 20 naturally occurring ROMK mutations with consecutive classification into mutational classes and identification of distinct rescue mechanisms according to the underlying defect. METHODS: Mutated ROMK potassium channels were expressed in Xenopus oocytes and a human kidney cell line and analyzed by two electrode voltage clamp analysis, immunofluorescence, and Western blot analysis. RESULTS: We identified 14 out of 20 ROMK mutations that did not reach the cell surface, indicating defective membrane trafficking. High expression levels rescued six out of 14 ROMK mutants, leading to significant K+ currents. In addition, two early inframe stop mutations could be rescued by aminoglycosides, resulting in full-length ROMK and correct trafficking to the plasma membrane in a subset of transfected cells. CONCLUSION: In contrast to previous reports, most of the investigated ROMK mutations displayed a trafficking defect that might be rescued by pharmacologic agents acting as molecular chaperones. The evaluation of different disease causing mechanisms will be essential for establishing new and more specific therapeutic strategies for HPS/aBS patients. PMID- 12911543 TI - Vasopressin-independent renal urinary concentration: increased rBSC1 and enhanced countercurrent multiplication. AB - BACKGROUND: A close association between the expression of the sodium transporter, rat bumetanide sensitive cotransporter (rBSC), in thick ascending limb of Henle and urinary concentration has been reported. However, direct evidence for this association and the mechanism of rBSC1 expression are still to be elucidated. METHODS: Brattleboro (BB) rats weighing approximately 200 g were dehydrated by water restriction for 4 hours, which induced around a 5% body weight reduction. Although plasma arginine vasopressin (AVP) was undetectable even after the water restriction, BB rats concentrated urine from 182 +/- 23 (mean +/- SD) at baseline to 404 +/- 65 mOsm/kg. H2O. RESULTS: Urinary volume was reduced from 5.8 +/- 1.8 to 1.4 +/- 0.6 mL/h. This treatment significantly increased sodium and urea accumulation in the renal medulla and reduced urinary sodium excretion. rBSC1 signals for both mRNA and protein were increased in dehydrated rats, although aquaporin type 2 (AQP2) expression was not enhanced in dehydrated BB rats. Subcutaneous infusion of desmopressin acetate (DDAVP) intensified rBSC1 signals of BB rats more than those in dehydrated condition. CONCLUSION: Dehydration increased rBSC1 expression and enhanced countercurrent multiplication even in AVP deficiency. These results supply strong evidence for the association between rBSC1 expression and urinary concentration, and indicate the presence of an AVP independent mechanism for urine concentration. PMID- 12911544 TI - Angiotensin II stimulates NHE3 activity by exocytic insertion of the transporter: role of PI 3-kinase. AB - BACKGROUND: Low-concentration angiotensin II (Ang II) stimulates Na+/H+ exchanger 3 (NHE3) activity in renal proximal tubule mainly via angiotensin II type 1 (AT1) receptors. The mechanisms that mediate the increase in NHE3 activity elicited by Ang II remain incompletely settled. METHODS: To assess a potential role of NHE3 trafficking in the Ang II effect, NHE3 activity was measured by H+-driven initial rate of 22Na uptake resistant to 50 micromol/L of the Na+/H+ exchange inhibitor cariporide (HOE642), and sensitive to 300 micromol/L ethyl isopropyl amiloride (EIPA), in a model of cultured proximal tubular cells (MKCC), in which functional apical NHE3 and AT receptors are normally present. Apical expression of NHE3 protein was determined by cell surface biotinylation and immunoblotting. RESULTS: Ang II (10-10 mol/L, 43 minutes) increased NHE3 activity and biotinylated NHE3 protein without any change in total amount of NHE3 protein. Both effects were suppressed by specific AT1 receptor antagonists. When 2-mercaptoethanesulphonic acid (MESNA) was used to cleave biotin from all apical proteins, intracellular biotinylated NHE3 protein remained unchanged after Ang II incubation compared to control. When sulfo-N-hydrosuccinimide (NHS)-acetate was used first to block all apical reactive sites, an increase in biotinylated NHE3 protein was observed following Ang II incubation. To evaluate the role of phosphatidylinositol 3 kinase (PI 3-kinase), the specific inhibitor wortmannin was used. It suppressed Ang II-induced increase in NHE3 activity and trafficking. Furthermore, latrunculin B, inhibitor of actin filament polymerization, prevented both Ang II stimulatory effects. CONCLUSION: Ang II stimulates NHE3 activity, at least in part, by exocytic insertion of the protein into the apical membrane. This effect is mediated by PI 3-kinase and required integrity of actin cytoskeleton. PMID- 12911545 TI - Vitamin E prevents renal dysfunction induced by experimental chronic bile duct ligation. AB - BACKGROUND: The mechanisms by which prolonged cholestasis alters renal hemodynamics and excretory function are unknown but may be related to increased oxidative stress, with subsequent formation of lipid peroxidation-derived products (e.g., F2-isoprostanes) and endothelin (ET). We investigated whether antioxidant therapy prevents chronic bile duct ligation (CBDL)-induced alterations in systemic and renal hemodynamics, and reduces F2-isoprostane and ET levels. METHODS: Sprague-Dawley rats were placed on either a normal or a high vitamin E diet for 7 days and then underwent either CBDL or sham surgery. They were then maintained on their respective diets for 21 more days, at which time the physiologic studies were performed. RESULTS: Thirty-three percent of the CBDL rats died by day 21. The remaining rats had a lower mean arterial pressure (MAP), renal blood flow (RBF), glomerular filtration rate (GFR), and sodium and water excretion than control rats. CBDL rats had higher portal pressure, renal venous pressure, and renal vascular resistance (RVR). These changes were associated with increased levels of systemic and renal venous F2-isoprostanes and ET. Vitamin E normalized MAP, RBF, GFR, RVR, and sodium and water excretion, and improved the 21-day survival without altering portal or renal venous pressures. Surprisingly, vitamin E did not alter the systemic levels of F2-isoprostanes but markedly reduced their levels in the renal venous circulation. CONCLUSION: Vitamin E improves MAP and renal function in CBDL rats, and selectively decreases renal levels of oxidative stress and ET, suggesting that local redox balance is implicated in CBDL-induced renal dysfunction. PMID- 12911546 TI - Endothelin-1 receptor blockade prevents renal injury in experimental hypercholesterolemia. AB - BACKGROUND: The potent vasoconstrictor endothelin-1 is involved in regulation of renal function, and is up-regulated in hypercholesterolemia (HC), a risk factor for renal disease that increases oxidative stress and impairs renal hemodynamic responses. However, the involvement of endothelin (ET) in this disease process is yet unknown. METHODS: Regional renal hemodynamics and function in vivo were quantified in pigs at baseline and during infusion of acetylcholine using electron beam computed tomography after a 12-week normal diet (N = 6), HC diet (N = 6), and HC diet orally supplemented (4 mg/kg/day) with the selective ET receptor-A (ET-A) blocker ABT-627 (HC+ET-A, N = 6). Plasma levels of 8-epi-PGF2 alpha-isoprostanes, markers of oxidative stress, were measured using enzyme immunoassay, and renal tissue was studied ex vivo using Western blotting, electrophoretic mobility shift assay, and immunohistochemistry. RESULTS: Total and low-density lipoprotein (LDL) cholesterol were similarly increased, but isoprostanes were decreased in HC+ET-A compared to HC alone. Basal renal perfusion was similar among the groups, while glomerular filtration rate (GFR) increased in HC+ET-A compared to HC. Stimulated perfusion and GFR were blunted in HC, but normalized in HC+ET-A. Moreover, ET blockade increased expression of endothelial nitric oxide synthase, and decreased endothelial expression of the oxidized-LDL receptor LOX-1, as well as tubular immunoreactivity of inducible nitric oxide synthase, nitrotyrosine, nuclear factor-kappaB, transforming growth factor-beta, and tubulointerstitial and perivascular trichrome staining. CONCLUSION: ET-A blockade improves renal hemodynamic and function in HC, and decreases oxidative stress, and renal vascular and tubulointerstitial inflammation and fibrosis. These findings support a role for the endogenous ET system in renal injury in HC and atherosclerosis. PMID- 12911547 TI - Angiotensin II regulates 11beta-hydroxysteroid dehydrogenase type 2 via AT2 receptors. AB - BACKGROUND: In preeclampsia, cortisol degradation by the enzyme 11beta hydroxysteroid dehydrogenase type 2 (11beta-HSD2) is compromised, which enhances intracellular cortisol availability. This leads to vasoconstriction and renal sodium retention with volume expansion, thus increasing blood pressure. An augmented availability of angiotensin II (Ang II) predisposes to preeclampsia. Some effects of Ang II are mediated by the mitogen-activated protein kinase (MAPK) cascade, which also regulates 11beta-HSD2 activity. Therefore, we hypothesized that Ang II regulates 11beta-HSD2. METHODS: The human choriocarcinoma cell line JEG-3, which expresses the 11beta-HSD2 isoenzyme, was used. 3H-cortisol/cortisone conversion assays and mRNA analyses by reverse transcription-polymerase chain reaction (RT-PCR) were performed. Cells were stimulated with Ang II and the effect was modulated by Ang II type 1 (AT1) and AT2 receptor blockers DUP753 or L-158809 and PD-123319, respectively. In order to elucidate the signaling cascade, the MAPK kinase inhibitors PD-098059 and U-0126 were probed. The impact of a modulated 11beta-HSD2 activity was assessed by determining the effect of cortisol on AT1 receptor mRNA. RESULTS: Ang II reduced mRNA and activity of 11beta-HSD2 mainly by a post-transcriptional mechanism. This Ang II effect was abrogated by AT2, but not by AT1 receptor blockade. Mitogen activated protein (MAP) kinase kinase (MAPKK) inhibitors reversed the Ang II effect. Dexamethasone augmented the mRNA expression of AT1 receptors. Cortisol enhanced AT1 receptor mRNA expression when the 11beta-HSD2 activity was reduced either by Ang II or by glycyrrhetinic acid, an 11beta-HSD2 inhibitor. CONCLUSION: Ang II decreases the activity of 11beta-HSD2 by an AT2 receptor- and MAPK dependent mechanism. The decreased activity of 11beta-HSD2 increases the intracellular availability of cortisol, which might be relevant for the pathogenesis of hypertension and preeclampsia. PMID- 12911548 TI - Alpha and gamma tocopherol metabolism in healthy subjects and patients with end stage renal disease. AB - BACKGROUND: The metabolism of alpha and gamma tocopherol, the major components of vitamin E, have not been studied in uremic patients. The major pathway of tocopherol metabolism is via phytyl side chain oxidation, leaving carboxyethyl hydroxychromans (CEHC) as metabolites. Alpha and gamma CEHC are water soluble, renally excreted, with known potent anti-inflammatory and antioxidative properties. METHODS: We examined serum alpha and gamma tocopherol and respective CEHC concentrations in 15 healthy subjects and 15 chronic hemodialysis patients. RESULTS: Serum alpha tocopherol levels were similar in hemodialysis patients (12.03 +/- 1.34 microg/mL) and healthy subjects (11.21 +/- 0.20 microg/mL), while serum gamma tocopherol levels were significantly greater in hemodialysis patients (3.17 +/- 0.37 microg/mL) compared to healthy subjects (1.08 +/- 0.06 microg/mL, P < 0.0001). Serum alpha and gamma CEHC levels were tenfold and sixfold higher in hemodialysis patients compared to healthy subjects, respectively (both P < 0.0001). Serum alpha and gamma tocopherol levels and CEHC metabolites were also measured after supplementation of alpha- or gamma-enriched mixed tocopherols in both hemodialysis patients and healthy subjects. Tocopherol administration resulted in modest or nonsignificant changes in serum tocopherol concentrations, while markedly increasing serum CEHC concentrations in both healthy subjects and hemodialysis patients. Hemodialysis resulted in no change in the serum alpha or gamma tocopherol concentrations while decreasing serum alpha CEHC and gamma CEHC levels by 63% and 53%, respectively (both P = 0.001 versus predialysis). Fourteen day administration of gamma-enriched but not alpha tocopherols lowered median C reactive protein (CRP) significantly in hemodialysis patients (4.4 to 2.1 mg/L, P < 0.02). CONCLUSION: First, serum alpha and gamma CEHC accumulate in uremic patients compared to healthy subjects; second, supplementation with tocopherols dramatically increases serum CEHC levels in both healthy subjects and hemodialysis patients; and, finally, CEHC accumulation may mediate anti inflammatory and antioxidative effects of tocopherols in hemodialysis patients. PMID- 12911549 TI - Percutaneous maxacalcitol injection therapy regresses hyperplasia of parathyroid and induces apoptosis in uremia. AB - BACKGROUND: A high level of parathyroid hormone (PTH) is considered to be an indicator of poor prognosis and a poor quality of life of dialysis patients; therefore, an effective and safe therapy for secondary hyperparathyroidism (SHPT) has been developed. METHODS: In 20 patients with SHPT resistant to maxacalcitol (OCT) intravenously administered, all detectably enlarged parathyroid glands were treated by percutaneous maxacalcitol injection therapy (PMIT) under ultrasonographic guidance consecutively 6 times, which was followed by OCT that was intravenously administered. The clinical effects of PMIT were evaluated based on the changes in the serum intact-PTH, adjusted Ca, phosphorus, and bone marker levels, and the parathyroid gland volume determined by ultrasonography. Morphologic examination, apoptosis analysis, and PTH mRNA expression level determination by reverse transcription-polymerase chain reaction (RT-PCR) using parathyroid tissues obtained by a biopsy technique were performed. RESULTS: PMIT and subsequent intravenous OCT administrations significantly decreased the serum intact-PTH level and parathyroid gland volume for at least 12 weeks after PMIT without major complications. Parathyroid tissues obtained after PMIT exhibited some partial defects of parathyroid cells, a marked increase in the number of the terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) positive cells, the ladder formation determined by DNA electrophoresis, and the decrease in the PTH mRNA expression level. CONCLUSION: PMIT is effective and safe for the treatment of refractory SHPT, and a locally high level of OCT suppresses PTH secretion and regresses parathyroid hyperplasia, which is involved in the induction of apoptosis of parathyroid cells. PMID- 12911550 TI - The quality of life and cost utility of home nocturnal and conventional in-center hemodialysis. AB - BACKGROUND: Home nocturnal hemodialysis is an intensive form of hemodialysis, where patients perform their treatments at home for about 7 hours approximately 6 nights a week. Compared with in-center conventional hemodialysis, home nocturnal hemodialysis has been shown to improve physiologic parameters and reduce health care costs; however, the effects on quality of life and cost utility are less clear. We hypothesized that individuals performing home nocturnal hemodialysis would have a higher quality of life and superior cost utility than in-center hemodialysis patients. METHODS: Home nocturnal hemodialysis patients and a demographically similar group of in-center hemodialysis patients from a hospital without a home hemodialysis program underwent computer-assisted interviews to assess their utility score for current health by the standard gamble method. RESULTS: Nineteen in-center hemodialysis and 24 home nocturnal hemodialysis patients were interviewed. Mean annual costs for home nocturnal hemodialysis were about 10,000 dollars lower for home nocturnal hemodialysis (55,139 dollars +/- 7651 dollars for home nocturnal hemodialysis vs. 66,367 dollars +/- 17,502 dollars for in-center hemodialysis, P = 0.03). Home nocturnal hemodialysis was associated with a higher utility score than in-center hemodialysis (0.77 +/- 0.23 vs. 0.53 +/- 0.35, P = 0.03). The cost utility for home nocturnal hemodialysis was 71,443 dollars/quality-adjusted life-year (QALY), while for in-center hemodialysis it was 125,845 dollars/QALY. Home nocturnal hemodialysis was the dominant strategy, with an incremental cost-effectiveness ratio (ICER) of -45,932 dollars. The 95% CI for the ICER, and 2500 bootstrap iterations of the ICER all fell below the cost-effectiveness ceiling of 50,000 dollars. The net monetary benefit of home nocturnal hemodialysis ranged from 11,227 dollars to 35,669 dollars. CONCLUSION: Home nocturnal hemodialysis is associated with a higher quality of life and a superior cost utility when compared to in-center hemodialysis. PMID- 12911551 TI - Expression of TGF-beta-induced matrix protein betaig-h3 is up-regulated in the diabetic rat kidney and human proximal tubular epithelial cells treated with high glucose. AB - BACKGROUND: betaig-h3 is an extracellular matrix protein whose expression in several cell types is greatly increased by transforming growth factor-beta (TGF beta). TGF-beta is believed to be involved in the development of diabetic nephropathy and thus we have assessed the possibility that betaig-h3 may be a downstream molecule in this pathogenic process. METHODS: Immunoblotting and immunohistochemistry were done using an antibody against mouse betaig-h3 protein. betaig-h3 and TGF-beta concentrations were measured by enzyme-linked immunosorbent assay (ELISA). Cell adhesion and migration were assessed by measuring activity of N-acetyl-beta-d-glucosaminidase and using a transwell plate, respectively. RESULTS: Immunohistochemistry revealed that betaig-h3 occurs mainly in the basement membrane of proximal tubules, particularly the S3 segment but also to lesser extents in the basement membranes of the cortical thick ascending limb cells and the parietal glomerular epithelial cells in Bowman's capsule. Immunoblotting revealed that approximately 68 kD bands were seen only in the cortex + the outer stripe of the outer medulla. Rats with streptozotocin (STZ)-induced diabetes exhibited a marked and sustained increase in renal betaig h3 abundance. This was mirrored by urinary betaig-h3 levels. In vitro experiments with human primary renal proximal tubular epithelial cells revealed that their expression of betaig-h3 was greatly increased by either TGF-beta or glucose. High glucose levels also stimulated TGF-beta production by renal proximal tubular epithelial cells and the high glucose-induced betaig-h3 expression was almost completely blocked by anti-TGF-beta antibody. betaig-h3 mediated renal proximal tubular epithelial cells adhesion and migration. CONCLUSION: betaig-h3 may be important in the development of diabetic nephropathy. Furthermore, the level of urinary betaig-h3 may be useful as an early marker reflecting disease onset and progression. PMID- 12911552 TI - The association between gout and nephrolithiasis in men: The Health Professionals' Follow-Up Study. AB - Approximately 10 million adults in the United States have experienced the passage of a kidney stone, and up to 5 million have been diagnosed with gout by a physician. Previous reports have suggested that gout increases the risk for the development of kidney stones, but there are no prospective data. We used data from a cohort of 51,529 male health care professionals to examine the independent association between gout and kidney stone disease. In a cross-sectional analysis of gout and kidney stone disease reported on the 1986 baseline questionnaire, the prevalence of kidney stone disease was almost twofold higher in men with history of gout compared to those without (15% vs. 8%). After adjusting for age and body mass index (BMI), a history of gout remained significantly associated with kidney stone disease (OR 1.88; 95% CI 1.68 to 2.11). We then prospectively examined the risk of incident kidney stones in men with and without a confirmed diagnosis of gout after excluding men who reported a history of kidney stone disease or gout on the baseline questionnaire. A confirmed diagnosis of gout increased the multivariate relative risk of incident kidney stones (RR 2.12; 95% CI 1.22 to 3.68). In contrast, a history of kidney stone disease was not associated with increased risk of gout (RR 1.05; 95% CI 0.54 to 2.07). In conclusion, a history of gout independently increases the risk for incident kidney stones in men. Physicians should provide dietary counseling, such as increasing fluid intake and decreasing salt consumption, to subjects with gout in addition to other risk factors, such as family history of kidney stones, in order to decrease the likelihood of stone formation. PMID- 12911553 TI - Conversion from cyclosporine to azathioprine at three months reduces the incidence of chronic allograft nephropathy. AB - BACKGROUND: Conversion from cyclosporine to azathioprine after renal transplantation has been shown to be beneficial in terms of allograft function, cardiovascular risk factor profile, and the incidence of gout. A higher incidence of acute rejection, however, has also been reported and uncertainty still exists about the long-term outcome after conversion. We report on the extended follow-up of an open-label, randomized trial that examined conversion to azathioprine as early as three months after transplantation. METHODS: One hundred twenty-eight patients were enrolled in this single-center study. Three months after transplantation they were randomly assigned to continue cyclosporine treatment (N = 68), or they were converted to azathioprine (N = 60). The steroid dose was temporarily increased in the patients who were converted. RESULTS: Patient survival was not different in the two groups. Graft survival tended to be lower (64.7% vs. 76.5% at 15 years) in the cyclosporine continuation group (P = 0.14) when data were analyzed on an intention to treat basis. The graft survival of the patients that stayed on their assigned treatment was significantly higher in the azathioprine arm, starting at two years' post-transplantation. The glomerular filtration rate was significantly higher in the patients who were converted to azathioprine. More allograft biopsies were taken from patients remaining on cyclosporine for suspicion of cyclosporine-related nephrotoxicity and prompted a high rate of late conversions (19%). The relative risk of chronic allograft nephropathy was significantly higher in the group that continued cyclosporine [relative risk, 4.3 (95% CI, 1.4 to 12.9); P = 0.009]. Conversion to azathioprine reduced the need of blood pressure and lipid-lowering drugs. CONCLUSION: Conversion to a calcineurin inhibitor-free immunosuppressive regiment three months after renal transplantation improved allograft function, reduced the need of cardiovascular risk factor-controlling medication, and reduced the incidence of chronic allograft nephropathy. PMID- 12911554 TI - Renal structure in early autosomal-dominant polycystic kidney disease (ADPKD): The Consortium for Radiologic Imaging Studies of Polycystic Kidney Disease (CRISP) cohort. AB - BACKGROUND: Autosomal-dominant polycystic kidney disease (ADPKD) is characterized by gradual renal enlargement and cyst growth prior to loss of renal function. Standard radiographic imaging has not provided the resolution and accuracy necessary to detect small changes in renal volume or to reliably measure renal cyst volumes. The Consortium for Radiologic Imaging Studies in Polycystic Kidney Disease (CRISP) is longitudinally observing ADPKD individuals using high resolution magnetic resonance (MR) imaging to determine if change in renal and cyst volumes can be detected over a short period of time, and if they correlate with decline in renal function early in disease. METHODS: Standardization studies were conducted in phantoms and four subjects at each participating clinical center. After, in the full-scale protocol, healthy ADPKD individuals 15 to 45 years old with creatinine clearance>70 mL/min underwent standardized MR renal imaging, renal iothalamate clearance, comprehensive clinical evaluation, and determination of 24-hour urinary albumin and electrolyte excretion. Stereology was used from T1-weighted images to quantify renal volume, and region-growing thresholding was used from T2-weighted images to determine cyst volume. Renal structures were evaluated in relation to demographic, clinical, and biochemical variables using means/medians, standard deviations, and Pearson correlations. RESULTS: Reliability coefficients for MR renal and cyst volume measurements in phantoms were 99.9% and 89.2%, respectively. In the full-scale protocol, 241 ADPKD individuals (145 women and 96 men) were enrolled. Total renal, cyst, and % cyst volume were significantly greater in each decade group. Hypertensive individuals demonstrated greater renal, cyst, and % cyst volume than normotensive subjects. Age-adjusted renal (r = -0.31, P < 0.0001), cyst (r = -0.36, P < 0.0001), and % cyst volume (r = -0.35, P < 0.0001) were inversely related to glomerular filtration rate (GFR). Age-adjusted renal volume (r = 0.42, P < 0.0001), cystic (r = 0.39, P < 0.0001, and % cyst volume (r = 0.41, P < 0.0001) were related with urinary albumin excretion. CONCLUSION: MR measures of renal and cyst volume are reliable and accurate in patients with ADPKD. ADPKD is characterized by significant cystic involvement that increases with age. Structure (renal and cyst volume) and function (GFR) are inversely related and directly related with the presence of hypertension and urinary albumin excretion in individuals with normal renal function. PMID- 12911555 TI - Hydroxyethyl starch and change in renal function in patients undergoing coronary artery bypass graft surgery. AB - BACKGROUND: Several case reports and clinical lore have suggested that exposure to the colloid hydroxyethyl starch may impair renal function, but few studies have systematically addressed this issue, and several have produced conflicting results. We sought to study the question in a formal analysis of postoperative change in renal function in patients undergoing coronary artery bypass graft (CABG) surgery. METHODS: We identified 238 consecutive patients who underwent CABG surgery at a large academic medical center. Glomerular filtration rate (GFR) was estimated using the Cockroft-Gault formula at baseline as well as on postoperative days 3 and 5. Linear regression analysis was used to study the relation between changes in GFR and intraoperative hydroxyethyl starch administration. Multivariate models controlled for potential demographic, clinical, and surgery-related confounders. RESULTS: Hydroxyethyl starch use was independently associated with a reduction in GFR on both postoperative days 3 and 5, with GFR declining by 7.2 mL/min/1.73 m2 on day 3 per unit of hydroxyethyl starch administered (95% CI, 1.7 to 12.7; P = 0.012), and by 6.6 mL/min/1.73 m2 on day 5 (95% CI, 1.2 to 11.9; P = 0.018). CONCLUSION: Intraoperative use of hydroxyethyl starch may be associated with modest impairment in renal function in patients undergoing CABG surgery. Randomized clinical trials will be necessary to confirm these findings and to further investigate their clinical implications. PMID- 12911556 TI - Angiotensinogen gene variation and renoprotective efficacy of renin-angiotensin system blockade in IgA nephropathy. AB - BACKGROUND: Blockade of the renin-angiotensin system (RAS) is well documented to be renoprotective; however, not all patients with glomerulonephritis respond well to this therapy. The interindividual variation in response to the RAS blockade may be in part genetically determined, whereas the results have been controversial. METHODS: We investigated whether the therapeutic efficacy of angiotensin-converting enzyme (ACE) inhibitors and/or angiotensin receptor blocker on renal prognosis is modified by the angiotensinogen gene (AGT) polymorphism in immunoglobulin A nephropathy (IgAN). In total, 259 patients with histologically proven IgAN were analyzed for clinical manifestations, renal survival, and their associations with AGT A(-20)C and M235T. RESULTS: The renal prognosis of 110 patients, who received ACE inhibitors/angiotensin receptor blocker during their clinical course, was significantly better than those without ACE inhibitors/angiotensin receptor blockers despite higher blood pressures and heavier proteinuria. The Cox proportional hazards regression model showed an increased hazard ratio (HR) for urinary protein (more than 1.0 g/day) of 3.346 (P = 0.0001), hypertension of 1.949 (P = 0.01), deteriorated renal function of 3.040 (P < 0.0001), no ACE inhibitor/angiotensin receptor blocker administration of 2.725 (P = 0.0004), and the T235 and C(-20) haplotype of 1.608 (P = 0.0322). Only in patients carrying at least one M235 and A(-20) haplotype did the administration of ACE inhibitors/angiotensin receptor blockers have no significant effect on the prognosis of renal function (Kaplan-Meier, log rank test, chi2 = 0.700; P = 0.4028), whereas it was significant in patients who had other haplotypes of AGT (chi2 = 11.805; P = 0.0006). CONCLUSION: This study provides evidence that the M235T and A(-20)C genotype of AGT can influence the therapeutic efficacy of a RAS blockade on the renal survival in IgAN. PMID- 12911557 TI - Reduced kidney size in patients with simple renal cysts. AB - BACKGROUND: Renal cysts frequently occur in the absence of genetic diseases but their significance is unknown. Since multiple cysts develop in many patients with advanced renal disease, we tested the hypothesis that isolated cysts are associated with early nephron loss. METHODS: All inpatient and outpatient sonograms performed by the Renal Division since 1995 were reviewed and, after exclusion of duplicate studies, patients with genetic cystic disorders, complex cysts, hydronephrosis, peripelvic cysts, end-stage renal disease and transplanted kidneys, 2526 were selected for this study, of which 385 had one or more cysts. Maximum renal length was used as an indication of size, and renal function was estimated by serum creatinine concentration. RESULTS: Both right and left kidney length were significantly less in patients with cysts (P < 0.0001), independent of which kidney contained the cyst(s). Renal lengths were less in patients with multiple cysts as opposed to single cysts but not in patients with bilateral cysts as compared to unilateral cysts. Cysts were twice as frequent in solitary kidneys than in dual kidneys (P = 0.01). In outpatients matched for age and gender, those with cysts had a higher serum creatinine concentration but with borderline significance (P = 0.06). Multivariate analysis revealed that age, gender, and renal length were each independent variables and together accounted for one third of the incidence of cysts. CONCLUSION: Kidney size is reduced in patients with simple renal cysts noted on renal sonography, and cysts may be associated with reduced renal function. This suggests that isolated cysts are an indication of nephron loss in patients with renal abnormalities. PMID- 12911558 TI - Antiphospholipid antibodies as a risk factor for atherosclerotic events in renal transplant recipients. AB - BACKGROUND: Epidemiologic studies reported that antiphospholipid antibodies (APAs) were independent predictors of atherosclerotic events. We recently reported a high prevalence of APAs in renal transplant recipients. Nevertheless, the role of APAs on atherosclerotic events has not been prospectively studied in this high-risk population. METHODS: Participants in the study were 324 consecutive renal transplant recipients. Patients were enrolled between January 1996 and May 1998 and followed up until June 2002. RESULTS: The patients were followed for a mean duration of 62 +/- 26 months. Eighty seven (26.8%) patients exhibited APAs. We found a slight, but significant, correlation between total plasma homocysteine (tHcy) concentration and anticardiolipin (ACA) titers (r = 0.26; P = 0.036). Fifty six athersclerotic events (17.2%) occurred in 54 patients. Atherosclerotic events occurred more frequently in patients with APAs (33% vs. 9%; P = 0.0003) and ACAs levels were higher in patients who experienced atherosclerotic events (23.7 +/- 13.1 IU vs. 13.9 +/- 9.4 IU; P = 0.003). APAs were associated with an increased risk of atherosclerotic events (RR, 2.82; 95% CI, 1.17 to 5.31). Cox regression analysis also revealed that age above the median (RR, 5.21; 95% CI, 1.67 to 17.13), a previous history of cardiovascular disease (RR, 3.54; 95% CI, 1.57 to 10.43), hyperhomocysteinemia (RR, 4.01; 95% CI, 1.22 to 14.61), and current smoking (RR, 2.17; 95% CI, 1.01 to 6.72) were risk factors for atherosclerotic events. CONCLUSION: The presence of APAs is an independent cardiovascular risk factor in renal transplant recipients. Prevention trials are necessary to assess the efficacy and safety of anticoagulation therapy in transplant patients with APAs. PMID- 12911559 TI - Impact of dialysis modality on survival of new ESRD patients with congestive heart failure in the United States. AB - BACKGROUND: It is hypothesized, but not proven, that peritoneal dialysis might be the optimal treatment for end-stage renal disease (ESRD) patients with established congestive heart failure (CHF) through better volume regulation compared with hemodialysis. METHODS: National incidence data on 107,922 new ESRD patients from the Center for Medicare and Medicaid Services (CMS) Medical Evidence Form were used to test the hypothesis that peritoneal dialysis was superior to hemodialysis in prolonging survival of patients with CHF. Nonproportional Cox regression models evaluated the relative hazard of death for patients with and without CHF by dialysis modality using primarily the intent-to treat but also the as-treated approach. Diabetics and nondiabetics were analyzed separately. RESULTS: The overall prevalence of CHF was 33% at ESRD initiation. There were 27,149 deaths (25.2%), 5423 transplants (5%), and 3753 (3.5%) patients lost to follow-up over 2 years. Adjusted mortality risks were significantly higher for patients with CHF treated with peritoneal dialysis than hemodialysis [diabetics, relative risk (RR) = 1.30, 95% confidence interval (CI) 1.20 to 1.41; nondiabetics, RR = 1.24, 95% CI 1.14 to 1.35]. Among patients without CHF, adjusted mortality risk were higher only for diabetic patients treated with peritoneal dialysis compared with hemodialysis (RR = 1.11, 95% CI 1.02 to 1.21) while nondiabetics had similar survival on peritoneal dialysis or hemodialysis (RR = 0.97, 95% CI 0.91 to 1.04). CONCLUSION: New ESRD patients with a clinical history of CHF experienced poorer survival when treated with peritoneal dialysis compared with hemodialysis. These data suggest that peritoneal dialysis may not be the optimal choice for new ESRD patients with CHF perhaps through impaired volume regulation and worsening cardiomyopathy. PMID- 12911560 TI - Glycosaminoglycan and proteoglycan inhibit the depolymerization of beta2 microglobulin amyloid fibrils in vitro. AB - BACKGROUND: Although several kinds of evidence suggest that glycosaminoglycans (GAGs) and proteoglycans (PGs) may contribute to the development of beta2 microglobulin-related (Abeta2m) amyloidosis, the precise roles of these molecules for the development of Abeta2m amyloidosis are poorly understood. METHODS: We investigated the effects of GAGs and PGs on the depolymerization of Abeta2m amyloid fibrils at a neutral pH, as well as on the formation of the fibrils at an acidic pH in vitro, using fluorescence spectroscopy with thioflavin T and electron microscopy. RESULTS: Depolymerization of Abeta2m amyloid fibrils at pH 7.5 at 37 degrees C was inhibited dose-dependently by the presence of some GAGs (heparin, dermatan sulfate, or heparan sulfate) or PGs (biglycan, decorin, or keratan sulfate proteoglycan). Electron microscopy revealed that a significant amount of Abeta2m amyloid fibrils remained in the reaction mixture with some lateral aggregation. Second, when monomeric beta2m was incubated with aggrecan, biglycan, decorin, or heparin at pH 2.5 at 37 degrees C for up to 21 days, the thioflavin T fluorescence increased depending on dose and time. Electron microscopy revealed the formation of rigid and straight fibrils similar to Abeta2m amyloid fibrils in beta2m incubated with biglycan for 21 days. CONCLUSION: These results suggest that some GAGs and PGs could enhance the deposition of Abeta2m amyloid fibrils in vivo, possibly by binding directly to the surface of the fibrils and stabilizing the conformation of beta2m in the fibrils, as well as by acting as a scaffold for the polymerization of beta2m into the fibrils. PMID- 12911561 TI - High vibration perception threshold and autonomic dysfunction in hemodialysis patients with intradialysis hypotension. AB - BACKGROUND: Intradialysis hypotension is a common problem among hemodialysis patients. Some studies have shown that autonomic neuropathy could be a major cause of intradialysis hypotension, whereas others have not. Furthermore, whether there are parallel changes in the autonomic nervous system and somatic nerves remains unclear. METHODS: We investigated the autonomic and peripheral nervous functions of 12 chronic hemodialysis patients who suffered from intradialysis hypotension, and of 12 age- and sex-matched hemodialysis patients who had stable blood pressure during hemodialysis. We used spectral analysis of their heart rate variability and systemic vascular resistance to evaluate autonomic functions. Vibrameter and nerve conduction studies to assess peripheral nervous function were also performed. Low-frequency/high-frequency ratio power index was used as a surrogate of sympathovagal balance. RESULTS: The power index rose progressively in the control group and reached significantly high levels at hour 4 compared to the basal values (3.7 +/- 0.5 vs. 2.1 +/- 0.3; P < 0.05). However in the group prone to hypotension, the power index remained almost unchanged. In addition, their systemic vascular resistance was lower than that in the control group (13.7 +/- 1.8 vs. 22.3 +/- 2.6 Wood units; P < 0.05). The vibration perception thresholds of the index finger and great toe were significantly higher in the group prone to hypotension (4.7 +/- 0.7 vs. 2.2 +/- 0.3 vibration units and 3.1 +/- 0.4 vs. 1.5 +/- 0.2 vibration units, respectively; both P < 0.05). CONCLUSION: We found that more severe damage to autonomic and peripheral nervous system occurred in patients prone to hypotension. PMID- 12911562 TI - Effect of TCN2 776C>G on vitamin B12 cellular availability in end-stage renal disease patients. AB - BACKGROUND: Transcobalamin II is a serum protein that transports vitamin B12 from the intestine to the tissues. This complex, holo-transcobalamin II, may reflect vitamin B12 availability in the body. Conflicting data exist with regard to the effect of a polymorphism in the gene coding for transcobalamin II, TCN2 776C>G, on transcobalamin II levels in the general population, which in turn may affect holo-transcobalamin II, vitamin B12, as well as total homocysteine (tHcy) plasma levels. The effect of TCN2 776C>G on vitamin B12 cellular availability in dialysis patients is unknown. METHODS: We examined the effect of TCN2 776C>G on holo-transcobalamin II, vitamin B12, and tHcy plasma concentrations in 120 dialysis patients. RESULTS: Holo-transcobalamin II levels were normal or supranormal in all patients and showed a linear association with albumin (r = 0.205, P = 0.025) and with vitamin B12 (r = 0.778, P = 0.001), but not with age, creatinine, body mass index, tHcy, ln-tHcy, vitamin B6, plasma folate, and red blood cell folate concentration. TCN2 776C>G showed no effect on holo transcobalamin II, vitamin B12, and tHcy concentration [one-way analysis of variance (ANOVA), post-hoc Scheffe test]. Multiple linear regression analyses showed that albumin and B12 are independently associated with holo-transcobalamin II, whereas TCN2 776C>G and MTHFR 677C>T had no effect. Independent predictors of ln-tHcy included creatinine, red blood cell folate, and the MTHFR 677TT genotype. There was also an effect of the TCN2 776CC genotype on ln-tHcy levels in this multivariate analysis, however, that deserves cautious interpretation because there was no effect of TCN2 genotypes by ANOVA and Scheffe test [median ln-tHcy concentrations according to TCN2 genotypes (micromol/L): CC, 3.22; CG, 3.30; GG, 3.23]. CONCLUSION: TCN2 776C>G does not influence holo-transcobalamin II or vitamin B12 levels, and has no major effect on tHcy concentrations of end-stage renal disease patients. PMID- 12911563 TI - Transforming growth factor beta1 genotype polymorphisms determine AV fistula patency in hemodialysis patients. AB - BACKGROUND: In hemodialysis patients with an arteriovenous (AV) fistula, access failure is primarily due to fistula stenosis, which predisposes to thrombosis and subsequent access loss. The risk for access failure differs interindividually, an observation that is independent from vascular anatomy in a significant number of patients. Fistula stenosis is histologically characterized by intimal hyperplasia, which is induced by growth factors, among which transforming growth factor beta1 (TGF-beta1) is of major importance. The quantitative production of TGF-beta1 interindividually differs due to polymorphisms in the gene region encoding the signal sequence of the cytokine. We hypothesized that the TGF-beta1 genotype, by influencing the development of arteriovenous fistula stenosis, determines the risk for vascular access failure. METHODS: One hundred twenty patients who had undergone placement of an AV fistula for initiation of hemodialysis treatment were genotyped for the polymorphic bases at position +869 and +915 of the TGF-beta1 gene. The primary end-point was time from fistula placement to access failure. RESULTS: AV fistula patency was significantly associated with the TGF-beta1 genotype (P = 0.0046); patency was 62.4% and 81.2% after 12 months for TGF-beta1 high and intermediate producers, respectively. In contrast, AV fistula patency neither differed between diabetic and nondiabetic patients, nor between patients with and without manifest cardiovascular disease. CONCLUSION: Polymorphisms in the gene region encoding the signal sequence of TGF beta1 influence the risk for hemodialysis access failure. By inducing synthesis of extracellular matrix proteins, overproduction of TGF-beta1 may accelerate the development of intimal hyperplasia, resulting in fistula stenosis and subsequent access failure. PMID- 12911564 TI - Anthropometrically estimated total body water volumes are larger than modeled urea volume in chronic hemodialysis patients: effects of age, race, and gender. AB - BACKGROUND: The modeled volume of urea distribution (Vm) in intermittently hemodialyzed patients is often compared with total body water (TBW) volume predicted from population studies of patient anthropometrics (Vant). METHODS: Using data from the HEMO Study, we compared Vm determined by both blood-side and dialysate-side urea kinetic models with Vant as calculated by the Watson, Hume Weyers, and Chertow anthropometric equations. RESULTS: Median levels of dialysate based Vm and blood-based Vm agreed (43% and 44% of body weight, respectively). These volumes were lower than anthropometric estimates of TBW, which had median values of 52% to 55% of body weight for the three formulas evaluated. The difference between the Watson equation for TBW and modeled urea volume was greater in Caucasians (19%) than in African Americans (13%). Correlations between Vm and Vant determined by each of the three anthropometric estimation equations were similar; but Vant derived from the Watson formula had a slightly higher correlation with Vm. The difference between Vm and the anthropometric formulas was greatest with the Chertow equation, less with the Hume-Weyers formula, and least with the Watson estimate. The age term in the Watson equation for men that adjusts Vant downward with increasing age reduced an age effect on the difference between Vant and Vm in men. CONCLUSION: The findings show that kinetically derived values for V from blood-side and dialysate-side modeling are similar, and that these modeled urea volumes are lower by a substantial amount than anthropometric estimates of TBW. The higher values for anthropometry-derived TBW in hemodialyzed patients could be due to measurement errors. However, the possibility exists that TBW space is contracted in patients with end-stage renal disease (ESRD) or that the TBW space and the urea distribution space are not identical. PMID- 12911565 TI - Anemia in pediatric hemodialysis patients: results from the 2001 ESRD Clinical Performance Measures Project. AB - BACKGROUND: Despite improvements in dialysis care, anemia remains a problem in pediatric hemodialysis patients. METHODS: To assess possible explanations for the anemia, clinical data were obtained from the Centers for Medicare and Medicaid Services on all hemodialysis patients ages 12 to <18 years between October and December 2000. Complete data were available for 435 of the 516 patients (84%). RESULTS: A total of 160 (37%) patients had a mean hemoglobin of <11 g/dL (anemic). The mean (+/- SD) age for these patients was 15.5 +/- 1.8 years compared to 15.9 +/- 1.5 years for the target hemoglobin patients (P < 0.05). Mean time on chronic dialysis was similar for both the anemic and target hemoglobin patients (>/=100 g/dL) ( approximately 3 years) but patients on dialysis <6 months were more likely to be anemic (67%). While nearly all patients were treated with erythropoietin, anemic patients received greater weekly erythropoietin doses (intravenous, anemia 374 +/- 232 units/kg/week vs. target hemoglobin 246 +/- 196 units/kg/week, P < 0.001; and subcutaneous, 304 +/- 238 units/kg/week vs. 167 +/- 99 units/kg/week, P < 0.05). A total of 59% of anemic patients had a mean transferrin saturation (TSAT) >/=20% compared to 71% of patients with a target hemoglobin (P < 0.01). A mean serum ferritin >/=100 ng/mL was present in approximately two thirds of the anemic and target hemoglobin patients. Approximately 60% of all children were treated with intravenous iron. The mean Kt/V values were lower for anemic patients (1.46 +/- 0.4 vs. 1.53 +/- 0.3, P < 0.05). Anemic patients were less likely to have a normal serum albumin (29% anemic vs. 52% target hemoglobin patients, P < 0.001). CONCLUSION: In the final multivariable regression model, dialyzing <6 months, a low albumin, and a mean TSAT <20% remained significant predictors of anemia in children. PMID- 12911567 TI - A blast from the mast? PMID- 12911566 TI - The impact of immune-activating processes following transplantation on chronic allograft nephropathy. AB - BACKGROUND: The long-term success of organ transplantation is influenced by numerous alloantigen-dependent and -independent risk factors. However, only very little information is presently available on the influence of systemic immune activating processes following organ engraftment. METHODS: To simulate the clinical situation of sequential organ transplantation, rat renal allograft recipients received additional immune activating stimuli (secondary donor specific and third-party skin grafts) after transplantation at serial time intervals (4 and 8 weeks). The overall observation period was 16 weeks. RESULTS: All control animals survived the observation period. In contrast, recipients receiving additional third-party or donor-specific skin grafts were beginning to die 12 weeks after organ engraftment with only few animals surviving 16 weeks. Systemic immune activation by additional third-party and in particular by additional donor-specific skin grafts resulted in significant temporary and long term functional deterioration. Morphologic changes progressed significantly, particularly after a secondary challenge with donor-specific skin grafts. ED1+ monocytes/macrophages, T-cell infiltrates, and intragraft mRNA expression for CD25 were significantly elevated by 16 weeks, following an additional immune challenge. Analysis of early intragraft events showed strong up-regulation of CD25 transcripts, suggesting fast stimulation of intragraft immune processes. CONCLUSION: Both alloantigen-specific and -unspecific systemic immune activation processes, following experimental organ transplantation, contribute to chronic graft deterioration. Those results seem relevant for long-term immunosuppressive protocols and clinical situations of sequential organ transplantation. PMID- 12911569 TI - Reverse epidemiology and obesity in maintenance dialysis patients. PMID- 12911568 TI - How to improve the early diagnosis of Fabry's disease? PMID- 12911571 TI - Mycophenolate mofetil (MMF) therapeutic approach in patients with chronic glomerulonephritis (GN). PMID- 12911572 TI - Impact of thyroid dysfunction on serum cystatin C. PMID- 12911573 TI - Chronic renal disease: a growing problem. PMID- 12911574 TI - The rediscovery and isolation of TFPI. AB - Tissue factor pathway inhibitor (TFPI) is a multivalent Kunitz-type proteinase inhibitor that produces factor (F)Xa-dependent feedback inhibition of the factor VIIa/tissue factor (FVIIa/TF) catalytic complex that is responsible for the initiation of coagulation. Since 1985, when Rapaport and colleagues reported that the lipoprotein fraction of plasma contained a FXa-dependent inhibitor of FVIIa/TF, myriad articles have established its biochemical structure, its mechanism of action, and its physiological importance. This brief personal account reviews historical studies that established the existence of the inhibitor and the events that led to its initial isolation. PMID- 12911575 TI - Genetic studies in complex disease: the case pro linkage studies. PMID- 12911576 TI - Genetic studies in complex disease: the case pro association studies. PMID- 12911577 TI - Angiogenesis and proteins of the hemostatic system. PMID- 12911578 TI - The role of plasminogen in angiogenesis in vivo. AB - Plasminogen, by virtue of its role in the degradation of extracellular matrix proteins and by facilitation of cell migration, may contribute to angiogenesis. OBJECTIVE: the purpose of this study was to evaluate the contribution of plasminogen to angiogenesis in vivo. METHODS: Angiogenesis was assessed in gene targeted mice with deficiencies of plasminogen, urokinase plasminogen activator (uPA), and urokinase receptor (uPAR) in a mouse corneal model. In wild-type mice, female and young mice showed a trend toward increased angiogenesis compared to males and old mice. Because of this influence of age and gender on angiogenesis, young, female mice (6-13 weeks of age) were used for this study. RESULTS: In response to angiogenic stimulation by basic fibroblast growth factor (bFGF), uPA deficient mice exhibited a decrease in new vessel formation as reflected by vessel length (0.47 in control vs. 0.33 mm in uPA-/- mice, P = 0.043), but new vessel formation was not altered (P = 0.107) in the uPAR deficient mice compared to control mice. A significantly decreased angiogenic response of new vessel formation to both vascular endothelial growth factor (VEGF) (P < 0.02) and bFGF (P < 0.007) was observed in Plg deficient (Plg-/-) mice (VEGF - 0.36 mm, bFGF - 0.67 mm) compared to Plg+/+ mice (VEGF - 0.56 mm, bFGF - 0.85 mm). CONCLUSIONS: These results demonstrate the importance of plasminogen, as well as of uPA, in angiogenesis in vivo. PMID- 12911579 TI - Functional characterization of transcription factor binding sites for HNF1-alpha, HNF3-beta (FOXA2), HNF4-alpha, Sp1 and Sp3 in the human prothrombin gene enhancer. AB - BACKGROUND: Prothrombin is a key component in blood coagulation. Overexpression of prothrombin leads to an increased risk of venous thrombosis. Therefore, the study of the transcriptional regulation of the prothrombin gene may help to identify mechanisms of overexpression. OBJECTIVES: The aim of our study was to localize the regions within the prothrombin enhancer responsible for its activity, to identify the proteins binding to these regions, and to establish their functional importance. METHODS: We constructed a set of prothrombin promoter 5' deletion constructs containing the firefly luciferase reporter gene, which were transiently transfected in HepG2, HuH7 and HeLa cells. Putative transcription factor (TF) binding sites were evaluated by electrophoretic mobility shift assays. The functional importance of each TF binding site was evaluated by site directed mutagenesis and transient transfection of the mutant constructs. RESULTS: We confirmed the major contribution of the enhancer region to the transcriptional activity of the prothrombin promoter. Analysis of this region revealed putative binding sites for hepatocyte nuclear factor HNF4, HNF3 beta and specificity protein(Sp)1. We identified six different TFs binding to three evolutionary conserved sites in the enhancer: HNF4-alpha (site 1), HNF1 alpha, HNF3-beta and an as yet unidentified TF (site 2) and the ubiquitously expressed TFs Sp1 and Sp3 (site 3). Mutagenesis studies showed that loss of binding of HNF3-beta resulted in a considerable decrease of enhancer activity, whereas loss of HNF4-alpha or Sp1/Sp3 resulted in milder reductions. CONCLUSIONS: The prothrombin enhancer plays a major role in regulation of prothrombin expression. Six different TFs are able to bind to this region. At least three of these TFs, HNF4-alpha, HNF3-beta and Sp1/Sp3, are important in regulation of prothrombin expression. PMID- 12911580 TI - The role of inflammation in vascular injury and repair. AB - Inflammation plays a critical role in the vascular response to injury. In particular, mechanical injury using techniques such as balloon angioplasty and stenting results in complex inflammatory reactions which influence proliferation of vessel wall constituents such as endothelial cells, smooth muscle cells, and extracellular matrix proteins. Inflammatory cells are recruited to the injured vessel wall initially as a reparative mechanism; however, these same inflammatory processes are also pivotal in the development of restenotic lesions. Leukocytes serve as the primary inflammatory cells but we now know that platelets produce a number of important inflammatory mediators. This review describes the mechanisms that regulate endothelial cell migration, smooth muscle cell activation, and extracellular matrix protein production, all of which are key components in the inflammatory response to vascular injury. PMID- 12911581 TI - Aspirin resistance: definition, mechanisms and clinical read-outs. PMID- 12911582 TI - Increased clearance of von Willebrand factor antigen post-DDAVP in Type 1 von Willebrand disease: is it a potential pathogenic process? AB - The mechanism of von Willebrand factor (VWF) clearance is not fully understood. The factors that affect VWF clearance, and the normal in vivo mechanism of clearance, may be relevant to the pathogenesis of Type 1 von Willebrand disease (VWD), in which there is a partial deficiency of VWF. In order to investigate the clearance of VWF in Type 1 VWD, the current study assessed the half-life of VWF antigen (t(1/2) VWF:Ag) in Type 1 VWD patients and individuals with mild hemophilia A following the administration of 1-deamino-8-d-arginine vasopressin (DDAVP; desmopressin). To date 20 individuals have been assessed, 13 with Type 1 VWD and seven with mild hemophilia A. The median t(1/2) VWF:Ag in the Type 1 VWD and mild hemophilia A groups were 4.6 h and 9.5 h, respectively. The difference between the t(1/2) VWF:Ag for the two groups was significant, P < 0.02. Analysis of the data showed a correlation between the t(1/2) VWF:Ag and the baseline VWF:Ag level prior to administration of DDAVP: lower baseline VWF:Ag levels were associated with a shorter t(1/2) VWF:Ag. These data suggest that increased clearance of VWF may be the pathogenic mechanism in some cases of Type 1 VWD. PMID- 12911583 TI - Soluble P-selectin levels, P-selectin polymorphisms and cardiovascular disease. AB - P-selectin is a member of the selectin family of cell adhesion molecules which are important in the transient attachment of leukocytes to endothelial cells and platelets. A number of polymorphisms in the gene encoding P-selectin have been identified. Objectives were to investigate the relationship of soluble P (sP) selectin with P-selectin gene polymorphisms and coronary artery disease (CAD). Two hundred and forty-nine patients, with extent of CAD characterized by >or=50% stenosis in one or more coronary arteries, and 252 healthy controls were studied. Soluble P-selectin was significantly higher in the patients than controls after adjustment for age, sex and smoking [patients 49.8 (47.5-52.1) ng mL-1; controls 46.7 (44.5-49.1) ng mL-1, P = 0.03). There was no association of sP-selectin with myocardial infarction (MI) or presence of >or=50% stenosis. The -1817 T/C, -1969 G/A and -2123 C/G (but not the Thr715Pro) polymorphisms were in strong linkage disequilibrium. The Thr715Pro polymorphism was significantly associated with sP selectin even after adjustment for covariates [TT 48.9 (46.9-50.0) ng mL-1; TP + PP 40.7 (38.1-43.6) ng mL-1, P < 0.0001]. A significant interaction of Thr715Pro and smoking status was identified in the determination of sP-selectin levels. There was no significant association of genotype at any of the polymorphism in relation to MI or stenosis. The Thr715Pro polymorphisms is associated with plasma sP-selectin. This association is modulated by smoking, although the underlying mechanism remains unclear. PMID- 12911584 TI - Prevalence and risk factors of non-fatal venous thromboembolism in the active population of the VITA Project. AB - Cost-effective strategies for the identification of subjects at risk of venous thromboembolism (VTE) in the active population are still lacking. Our objectives were to identify risk factors for venous thromboembolism in active subjects. We analyzed data from a population-based sample of 15055 Caucasians aged 18-65 years randomly selected from the census list of the township of Vicenza, Italy. A validated methodology was used to retrospectively identify subjects with previous VTE. Body mass index (BMI), smoking, oral contraceptive use, previous superficial vein thrombophlebitis (SVT) and familial history of VTE, all at the age of first thrombosis, were ascertained by direct interview and by review of available medical records. Ninety-two deep vein thromboses [prevalence: 61.1/10000, 95% confidence interval (CI) 49.2-74.9], three upper deep vein thrombosis (prevalence: 1.9/10000, 95% CI 0.4-5.8) and 21 pulmonary embolism (prevalence: 13.9/10000, 95% CI 8.6-21.3) were identified. After age and sex adjustment, clinically identifiable risk factors were: history of SVT [odds ratio (OR) = 6.8], oral contraceptive use (OR = 4.7), family history of VTE (OR = 4.5), smoking (OR = 1.7) and BMI above the third tertile (OR vs. mid-tertile 2.9). While previous SVT and BMI were associated with VTE in all circumstantial situations (surgery/trauma, pregnancy or idiopathic VTE), for oral contraceptive use, positive family history and smoking the degree of association varied significantly depending on the situation. Non-fatal VTE affects 0.7% of the subjects belonging to an active population, 56% of cases being potentially preventable. In 30% of VTE cases, at least two easily recognizable risk factors are present. Clinical assessment of risk factors remains the mainstay of VTE prevention. PMID- 12911585 TI - Incidence of cancer after a first episode of idiopathic venous thromboembolism treated with 3 months or 1 year of oral anticoagulation. AB - BACKGROUND: A prolonged treatment with oral anticoagulants has been claimed to reduce the incidence of newly diagnosed cancer in the long-term follow-up of patients with venous thromboembolism. OBJECTIVES: In a multicenter prospective study we assessed the incidence of newly diagnosed clinically overt cancer in patients with a first episode of idiopathic venous thromboembolism (VTE) treated with oral anticoagulants for 3 months or 1 year. PATIENTS AND METHODS: Consecutive patients with an idiopathic venous thromboembolism who had completed 3 months of oral anticoagulant therapy without having a recurrence, bleeding or newly diagnosed cancer were randomized to discontinue oral anticoagulant therapy or to continue it for nine additional months. Idiopathic venous thromboembolism was defined as thrombosis occurring in the absence of known cancer, known thrombophilia, or temporary risk factors for venous thromboembolism. All patients were followed up for at least 1 year after randomization. RESULTS: A total of 429 patients, 265 patients with DVT and 164 with PE, were followed up for an average of 43.7 months after randomization. A newly diagnosed cancer occurred in 32 patients (7.5%), 13 (6.2%) of the 210 patients treated for 3 months and 19 (8.7%) of the 219 patients treated for 1 year (RR = 0.71, 95% confidence interval 0.36 1.41). CONCLUSIONS: The incidence of newly diagnosed clinically overt cancer is not reduced in patients with idiopathic venous thromboembolism treated with 1 year anticoagulant treatment compared with patients treated for 3 months. PMID- 12911586 TI - Alpha2-antiplasmin plays a significant role in acute pulmonary embolism. AB - The importance of pulmonary embolism (PE) due to venous thrombosis is recognized in the treatment of vascular diseases. We have investigated the physiological effects of plasmin generation in experimental acute PE using mice deficient in plasminogen (Plg-/-) or alpha2-antiplasmin (alpha2-AP-/-). PE was induced by continuous induction of venous thrombus in the left jugular vein by endothelial injury due to photochemical reaction. The mortality of wild-type mice was 68.8% at 2 h after the initiation of venous thrombosis and it was significantly reduced in alpha2-AP-/- mice (41.7%). In contrast, Plg-/- mice did not survive. Histological evidence of thromboembolism in the lung was obtained in all mice. However, whereas a strict thromboembolism was observed in Plg-/- mice, only a few thrombi were detected in the lungs of alpha2-AP-/- mice. Plasma fibrinogen levels measured in mice were not different. When alpha2-AP was infused in alpha2-AP-/- mice, the mortality was indistinguishable from wild-type mice. Tissue-type plasminogen activator (tPA) did not reduce the mortality due to acute PE in wild type mice. However, in alpha2-AP-/- mice, tPA (0.52 mg x kg-1) significantly decreased the mortality compared with that of alpha2-AP-/- mice without tPA. The bleeding time was not significantly prolonged in either type of mice treated with tPA. The lack of plasminogen increases the mortality due to acute PE while a lack of alpha2-AP decreases the mortality rate, which can be further reduced by tPA administration. Therefore, the combination of inhibition of alpha2-AP with thrombolytic therapy could be beneficial in the treatment of acute PE. PMID- 12911587 TI - Acenocoumarol therapy in pediatric patients. AB - To determine guidelines for administering and monitoring acenocoumarol therapy in children, 93 patients (median 5.1 years, range: 0.2-18 years) were prospectively evaluated over a 33-month period. The loading doses used were: <1 year, 0.20 mg x kg-1; >1-5 years, 0.09 mg x kg-1; 6-10 years, 0.07 mg x kg-1; 11-18 years, 0.06 mg x kg-1. In this study, the loading dose and the dose to achieve and maintain target therapeutic range (TTR) for acenocoumarol are age-dependent, with infants having the highest and teenagers having the lowest requirements. The use of a different loading dose according to age has allowed most of the children (80%) in all the age groups to achieve TTR in less than 1 week. No patients had serious bleeding or thrombotic complications. We conclude that there is an age-dependent response to acenocoumarol in pediatric patients. The implementation of an age adjusted loading dose regimen reduces the length of hospitalization required to achieve effective anticoagulant therapy. PMID- 12911588 TI - A high-score Mediterranean dietary pattern is associated with a reduced risk of peripheral arterial disease in Italian patients with Type 2 diabetes. AB - BACKGROUND: The 'Mediterranean diet' is considered to exert protective effects on cardiovascular disease, although a wide range of dietary patterns exists among subjects living even in the same Mediterranean country. OBJECTIVE: To investigate the association between specific dietary patterns and peripheral arterial disease (PAD) in Italian Type 2 diabetes patients. DESIGN: From a cohort of 944 patients with Type 2 diabetes, 144 patients with PAD were selected, and matched for age and sex with 288 Type 2 diabetic control patients without macrovascular complications. A dietary score was elaborated from a semiquantitative food frequency questionnaire. The higher the final score, the healthier the eating habit. RESULTS: In multivariate analysis, a higher score was independently associated with a significant reduction in PAD risk [odds ratio (OR) = 0.44; 95% confidence interval (CI) 0.24, 0.83]. Diabetes duration (OR > 15 years = 2.49; 95% CI 1.45, 4.25), hypertension (OR = 2.12; 95% CI 1.31, 3.45) and butter consumption (OR = 2.6; 95% CI 1.15, 3.68) were also significantly associated with PAD. The dietary score significantly improved the predictive value of models based on duration of diabetes and hypertension. (LSR = 2.19, DF = 7, P < 0.001). The effect of a high dietary score on the risk of PAD was independent of diabetes duration and hypertension. CONCLUSION: In Italian Type 2 diabetics, a higher dietary score has a protective role against PAD. The use of butter increases the risk of PAD even in patients regularly consuming olive oil. Dietary advice may be helpful for the prevention of PAD in diabetics even in populations traditionally accustomed to a Mediterranean dietary habit. PMID- 12911590 TI - Hepatic infarctions during pregnancy are associated with the antiphospholipid syndrome and in addition with complete or incomplete HELLP syndrome. AB - Antiphospholipid antibody syndrome (APS) is associated with adverse pregnancy outcomes and maternal complications including thrombotic events and early pre eclampsia. HELLP syndrome (Hemolysis, Elevated Liver enzymes, Low Platelets) represents a unique form in the spectrum of pre-eclampsia. This report describes four patients with pregnancy-associated hepatic infarctions. All four had APS and HELLP syndrome, which was complete in one patient and incomplete in three patients, with elevated liver enzymes in all, and either thrombocytopenia or hemolysis in two. In the literature, we found descriptions of an additional 24 patients who had 26 pregnancies with concomitant hepatic infarction. Of the total 28 patients, anticardiolipin antibodies (aCL) and/or lupus anticoagulant (LAC) were assessed in 16 patients, out of whom 15 were found to be positive. Hepatic infartction during pregnancy was associated almost always with APS, with HELLP (2/3 complete, 1/3 incomplete), and only in one-third of the pregnancies with pre eclampsia (PE). PMID- 12911589 TI - Decrease of hemostatic cardiovascular risk factors by aggressive vs. conventional atorvastatin treatment in patients with Type 2 diabetes mellitus. AB - BACKGROUND: Patients with Type 2 diabetes mellitus have increased levels of hemostatic risk variables for cardiovascular disease, such as fibrinogen, von Willebrand factor (VWF), factor (F)VIIa, d-dimer and plasminogen activator inhibitor-1 (PAI-1). OBJECTIVES: To evaluate the effect of aggressive vs. standard dose atorvastatin on hemostatic cardiovascular risk factors in patients with Type 2 diabetes mellitus. PATIENTS AND METHODS: The effect of 30 weeks of treatment with atorvastatin 10 and 80 mg on hemostatic cardiovascular risk factors was assessed in a randomized double-blind placebo-controlled trial on 217 patients with Type 2 diabetes mellitus and dyslipidemia. RESULTS AND CONCLUSIONS: Atorvastatin 10 and 80 mg dose-dependently reduced d-dimer (7.4% and 8.5%, respectively, P for trend = 0.004) and PAI-1 antigen levels (9.0% and 18%, respectively, P for trend = 0.021). Levels of fibrinogen, VWF, tissue-type plasminogen activator and FVIIa were not influenced by atorvastatin. In conclusion, in patients with Type 2 diabetes mellitus, atorvastatin dose dependently improved the levels of the hemostatic risk variables d-dimer and PAI 1. PMID- 12911591 TI - Autoimmune antiphospholipid antibodies impair the inhibition of activated factor X by protein Z/protein Z-dependent protease inhibitor. AB - The hemostatic process is tightly regulated by several antithrombotic mechanisms. Among them, protein Z (PZ)-dependent protease inhibitor (ZPI) potently inhibits factor (F)Xa in a manner dependent on calcium ions, phospholipids and PZ. Autoimmune antiphospholipid antibodies (aPL) are mainly directed against phospholipid-binding plasma proteins such as beta2-glycoprotein I (beta2GPI) and prothrombin, and are known to interfere with phospholipid-dependent hemostatic pathways. In this study, we investigated whether purified aPL are able to interfere with inhibition of FXa by PZ/ZPI. beta2GPI modestly delayed the FXa inactivation by PZ/ZPI and most isolated aPL-IgGs were found to further increase the inhibitory potential of beta2GPI on PZ/ZPI activity. Without beta2GPI, the PZ/ZPI activity was unaffected by the addition of aPL-IgG. As PZ deficiency is hypothesized to lead to a prothrombotic state, we performed a case-control study to measure plasma levels of PZ and ZPI in 66 patients with autoimmune aPL and 152 normal controls. The prevalence of low PZ levels (below the 5th percentile of controls) was significantly greater in the 37 patients with definite antiphospholipid syndrome (APS) (24.3%) but not in the 29 aPL patients not fulfilling the criteria for APS (10.3%) compared with the normal group (4.6%, P < 0.001 vs. APS). ZPI antigen levels were similar in patients with aPL and normal controls. Concomitant PZ deficiency increased by approximately sevenfold the risk of arterial thrombosis in aPL patients. Taken together, these data suggest that the PZ/ZPI system is commonly impaired in aPL patients thus probably increasing the thrombotic risk. PMID- 12911592 TI - Linkage analysis of factor VIII and von Willebrand factor loci as quantitative trait loci. AB - Elevated factor (F)VIII levels contribute to venous thrombotic risk. FVIII levels are determined to a large extent by levels of von Willebrand factor (VWF), its carrier protein which protects FVIII against proteolysis. VWF levels are largely dependent on ABO blood group. Subjects with blood group non-O have higher VWF and FVIII levels than individuals with blood group O. Apart from ABO blood group no genetic determinants of high FVIII levels have been identified, whereas clustering of FVIII levels has been reported within families even after adjustment for ABO blood group and VWF levels. We investigated the FVIII and VWF loci as possible quantitative trait loci (QTL) influencing FVIII and VWF levels. Two sequence repeats in the FVIII gene and three repeats in the VWF gene were typed in 52 FV Leiden families. Multipoint sib-pair linkage analysis was performed with the MAPMAKER/SIBS program. FVIII levels adjusted for VWF levels and age, and VWF levels adjusted for ABO blood group and age, were used for this linkage analysis. No linkage of FVIII levels to the FVIII locus was found, whereas we found evidence that the VWF locus contains a QTL for VWF levels [maximum likelihood no dominance variance lod score = 0.70 (P = 0.04) and non parametric Z-score = 1.92 (P = 0.03)]. About 20% of the total variation in VWF levels may be attributed to this VWF locus. PMID- 12911593 TI - Human CD4+ T-cell epitope repertoire on the C2 domain of coagulation factor VIII. AB - Approximately 25% of severe hemophilia A patients develop antibodies (Ab) that neutralize the procoagulant function of factor (F)VIII (inhibitors). Autoimmune FVIII inhibitors may develop in individuals without congenital FVIII deficiency and cause acquired hemophilia. Low titers of anti-FVIII Ab may be present in hemophilia A patients without inhibitors and in healthy blood donors. FVIII specific CD4+ T-cells drive the synthesis of anti-FVIII Ab. We examined the epitope repertoire of CD4+ T-cells from 15 healthy subjects, 10 hemophilia A patients without inhibitors, 11 hemophilia A patients with inhibitors, and six acquired hemophilia patients. Blood CD4+ T-cells were challenged in proliferation assays with a panel 16 overlapping synthetic peptides, spanning the sequence of the FVIII C2 domain. The sequence region 2291-2330 contained the most frequently and strongly recognized peptides in each of the four subject groups. Crystallographic B factor data and the location of these peptides within the three-dimensional structure of the C2 domain confirm that this region has a high degree of solvent exposure and flexibility within the peptide backbone, which are structural features typical of immunodominant universal CD4+ epitopes. Furthermore, this sequence region overlaps inhibitor-binding sites, suggesting that CD4+ T-cells recognizing peptide sequences within this region might be involved in inhibitor synthesis. The sequence regions 2191-2210 (recognized strongly by each study group except hemophilia A patients with inhibitors) and 2241-2290 (recognized primarily by acquired hemophilia patients and healthy subjects) share the same structural features, and also overlap inhibitor-binding sites. Although similar, there appear to be important differences in the CD4+ epitope repertoires of congenital and acquired hemophilia patients. PMID- 12911594 TI - Elevated thrombin-forming capacity of tissue factor-activated cord compared with adult plasma. AB - Clinically observed excellent hemostasis in neonates despite low levels of clotting factors is not completely understood so far. Therefore, we investigated whether physiological low levels of the inhibitor protein C (PC) facilitate thrombin formation in tissue factor (TF)-activated plasma samples. PC was activated by endogenously generated thrombin after addition of soluble thrombomodulin (TM). The capability of activated PC (APC) to suppress thrombin formation was significantly more pronounced in adult than in cord plasma. Addition of 4 nm of TM decreased the thrombin potential (TP) in cord plasma by 10%, and in adult plasma by 52% in the presence of 5 pm TF. We demonstrate that this low anticoagulant action of PC is attributable to the low levels of tissue factor pathway inhibitor (TFPI) and antithrombin (AT) physiologically present in cord plasma. Addition of 4 nm TM decreased the TP by 58% in cord plasma adjusted to contain TFPI and AT at adult levels in the presence of 5 pm TF. Thus, the combined low anticoagulant action of the three inhibitors APC, TFPI, and AT in cord plasma allows enhanced thrombin formation associated with shorter clotting times compared with adult plasma when low amounts of TF are applied to initiate clot formation. Although our laboratory experiments do not allow definite conclusions for various clinical situations, our data might contribute to explain excellent hemostasis in neonates despite low levels of procoagulants. PMID- 12911595 TI - Fine mapping of the sequences in domain 5 of high molecular weight kininogen (HK) interacting with heparin and zinc. AB - We previously localized the heparin binding region on high molecular weight kininogen to domain 5 (D5) by quantifying the binding using surface plasmon resonance of D5 fused at its N-terminal to glutathione-S-transferase. We further examined GST-(H475-S626) which at 100 nm was previously shown to be ineffective in reversing the heparin acceleration of antithrombin inhibition of thrombin. However, we now show that at a concentration of 400 nm, complete reversal of accelerated inhibition occurred. To characterize the interacting sequences on D5, four peptides representing surface loops of a molecular model were synthesized. Peptides H475-H485 and G440-G455, rich in histidine and low in lysine, showed weak or no detectable binding in the absence of Zn++, but tighter binding in the presence of Zn++. H483-K497 containing three histidines and six lysines showed tight binding without Zn++, and increased in avidity with Zn++. In contrast, G486 K502, low in histidine and high in lysine, showed tight binding (KD = 0.8 microm) in the absence and presence of Zn++. Both H483-K497 and G486-K502 were effective in neutralizing the accelerated inhibition by heparin of thrombin by antithrombin in the absence of Zn++. Therefore, a set of lysine residues in the sequence of K487-K502 is responsible for Zn++-independent binding of heparin. Further, a group of histidine residues in sequence range of H475-H485 contributes to Zn++ dependent binding of heparin to HK-D5. PMID- 12911596 TI - Lack of association of the plasminogen activator inhibitor-1 4G/5G promoter polymorphism with cardiovascular disease in the elderly. AB - Elevated circulating plasminogen activator inhibitor-1 (PAI-1) may increase risk of cardiovascular disease (CVD). The 4G allele of the 4G/5G PAI-1 promoter polymorphism is associated with higher levels of PAI-1. We examined the association of PAI-1 4G/5G genotype and CVD events in the elderly participants of the Cardiovascular Health Study (CHS). We measured 4G/5G genotype in a nested case-control study within the CHS. Cases included incident angina, myocardial infarction (MI), and stroke. 4G/5G genotype was not found to be associated with markers of fibrinolysis or CVD risk in the selected elderly cohort. There were no differences in genotype frequencies by case-control status (5G/5G frequency 16 22%; chi2P= 0.07). The 5G allele was not associated with incident CVD events when individuals with at least one 5G allele were compared to 4G/4G homozygotes. The presence of at least one 4G allele was likewise not associated with incident CVD when those with 4G/4G and 4G/5G genotypes were compared to 5G/5G homozygotes. Our results suggest that the PAI-1 4G/5G promoter polymorphism is not associated CVD risk factors or incident CVD events in the elderly. PMID- 12911597 TI - Glycoprotein IIb/IIIa inhibition attenuates platelet-activating factor-induced platelet activation by reducing protein kinase C activity. AB - Glycoprotein (GP)IIb/IIIa inhibition may abolish activated leukocyte-induced platelet activation, in which leukocyte-released platelet-activating factor (PAF) is a major mediator. The present study thus investigated if and how GPIIb/IIIa inhibitors interfere with PAF-induced platelet activation. Platelet and leukocyte activation were monitored by flow cytometry and immunoblotting. GPIIb/IIIa inhibitors (c7E3, non-peptide SR121566, and MAb RFGP56) attenuated PAF-induced, but not adenosine diphosphate (ADP)- or thrombin receptor activating peptide (TRAP)-induced platelet P-selectin expression in whole blood. GPIIb/IIIa blockade enhanced ADP- or TRAP-induced leukocyte CD11b expression, but not the response to PAF. GPIIb/IIIa blockade attenuated PAF-induced, but enhanced ADP- or TRAP induced platelet-leukocyte aggregation. Under the present experimental conditions, thromboxane A2 receptor antagonism did not significantly influence PAF-induced platelet activation, and GPIIb/IIIa inhibition did not interfere with calcium mobilization/influx in platelets. Protein kinase C (PKC) blockade inhibited PAF-induced platelet P-selectin expression, and PAF-induced PKC activity was reduced by GPIIb/IIIa inhibition. PAF (=1 micro m) did not induce MEK 1/2 or ERK 1/2 phosphorylation, whilst thrombin induced marked responses, which were enhanced by GPIIb/IIIa blockade. Thus, GPIIb/IIIa inhibition attenuates PAF-induced platelet activation via inhibiting PKC activity. GPIIb/IIIa blockade enhances thrombin-induced platelet MEK 1/2 and ERK 1/2 activation, and augments ADP- and TRAP-induced leukocyte activation by enhancing platelet-leukocyte aggregation. PMID- 12911598 TI - TRA-418, a novel compound having both thromboxane A(2) receptor antagonistic and prostaglandin I(2) receptor agonistic activities: its antiplatelet effects in human and animal platelets. AB - TRA-418 is a novel compound that has been found in our screening for compounds having both thromboxane A2 (TP) receptor antagonistic and prostaglandin I2 (IP) receptor agonistic activities. In the binding assays, TRA-418 showed a 10-fold higher affinity to TP-receptors than IP-receptors. TRA-418 inhibited platelet aggregation induced by the TP-receptor agonist, U-46619 and by arachidonic acid at concentrations lower than those required for inhibition of ADP-induced aggregations. Furthermore, TRA-418 inhibited not only platelet aggregation induced by ADP alone, but also that induced by ADP in the presence of the TP receptor antagonist, SQ-29548. When the IC50 values of TRA-418 for platelet aggregation were estimated in platelet preparations from monkeys, dogs, cats, and rats using ADP and arachidonic acid as the platelet stimulating agents, it was found that the values estimated in monkey platelets were quite similar to those estimated in human platelets. In ex vivo platelet aggregation in monkeys, TRA-418 exhibited significant inhibitory effects on arachidonic acid-induced aggregation in platelet preparations from monkeys treated at 3 micro g kg min-1 or higher doses, where neither a significant decrease in blood pressure nor a significant increase in heart rate was observed. These results are consistent with the fact that TRA-418 has a relatively potent TP-receptor antagonistic activity together with a relatively weak IP-receptor agonistic activity. PMID- 12911599 TI - Flavonoid inhibition of platelet procoagulant activity and phosphoinositide synthesis. AB - Dietary flavonoids are known for their antiplatelet activity resulting in cardiovascular protection. Phosphatidylinositol 4,5-bisphosphate (PIP2) was previously reported to play a direct role in phosphatidylserine (PS) exposure, as a Ca2+ target. Thrombin formation and platelet procoagulant activity are dependent on PS exposure. As flavonoids can inhibit phosphoinositide (PPI) kinases, we examined whether changes in PPI metabolism in flavonoid-treated platelets could be involved in their antiplatelet effects. Treatment with the flavonoids quercetin or catechin reduced PS exposure, thrombin formation, PIP2 level and resynthesis after platelet activation with collagen, thrombin or calcium ionophore. Flavonoids also prevented [Ca2+]i increase induced by collagen, but not by the ionophore. The ability of flavonoids to decrease PS exposure induced by ionophore treatment could result from the diminution of PIP2 levels, whereas PS exposure induced by collagen could also be diminished by flavonoids' effects on calcium signaling dependent on PIP2 hydrolysis. These data favor a role for PIP2 in the antiplatelet effects of flavonoids. PMID- 12911600 TI - Function-modulating human monoclonal antibodies against platelet-membrane receptors isolated from a phage-display library. AB - Monoclonal antibodies to platelet membrane receptors have been used extensively for analysis of receptor structure and function. Function-blocking human antibodies are being used for the development of antiplatelet drugs. We isolated human monoclonal antibodies from a library of single-chain Fv (scFv) antibodies displayed on the surface of filamentous phage, by selection on whole platelets. Eight different platelet-binding clones were isolated, of which three bound to the platelet-membrane glycoprotein (GP) GPIb in an ELISA assay. Specific elution with a recombinant polypeptide of von Willebrand factor (VWF) spanning the GPIbalpha binding site, yielded the same three phage clones. Two of the three anti-GPIb clones could be purified as scFv monoclonal antibodies, and they competed with each other for binding to intact platelets, suggesting that they bind at or near the same site on GPIb. Their binding affinities differed, however, and the clone with higher affinity inhibited ristocetin-induced platelet aggregation. These data indicate that selection from a phage display library of human scFvs using whole platelets can be applied for the isolation of functional antiplatelet-GPIb antibodies useful for the development of new therapeutic and diagnostic strategies. PMID- 12911601 TI - Hemophilia, rodentophilia and humanity. PMID- 12911602 TI - Anticoagulation with warfarin downregulates inflammation. PMID- 12911603 TI - Asymptomatic excessive coumarin anticoagulation is a risk factor for thrombotic and bleeding complications of oral anticoagulant therapy. PMID- 12911604 TI - Low physician compliance of prescribing anticoagulant prophylaxis in patients with solid tumor or hematological malignancies and central vein catheters. PMID- 12911605 TI - Does negative heparin-platelet factor 4 enzyme-linked immunosorbent assay effectively exclude heparin-induced thrombocytopenia? PMID- 12911606 TI - Increased plasma levels of soluble platelet glycoprotein V in patients with acute myocardial infarction. PMID- 12911607 TI - Cerebral vein thrombosis and right foot ischemia in a 21-year-old man. PMID- 12911608 TI - Identification and characterization of a natural R96C EPCR variant. PMID- 12911609 TI - Pattern of symptoms in 93 Iranian patients with severe factor XIII deficiency. PMID- 12911610 TI - Common single nucleotide polymorphisms in the promoter region of the human factor XI gene. PMID- 12911611 TI - No Val34Leu polymorphism of the gene for factor XIIIA subunit was detected by ARMS-RACE method in three Asian populations. PMID- 12911612 TI - Preserving eye function in prematurely born children with severe protein C deficiency. PMID- 12911614 TI - Tumor necrosis factor-alpha at the crossroads of neuronal life and death during HIV-associated dementia. AB - Human immunodeficiency type-1 (HIV-1) infection is known to cause disorders of the CNS, including HIV-associated dementia (HAD). It is suspected that tumor necrosis factor-alpha (TNF-alpha) released by infected microglia and macrophages play a role in neuronal injury seen in HAD patients. Accordingly, studies suggest that the level of TNF-alpha mRNA increases with increasing severity of dementia in patients, and that inhibitors of TNF-alpha release reduces neuronal injury in murine model of HAD. However, the exact role of TNF-alpha in relation to neuronal dysfunction is a matter of ongoing debate. One school of thought hails TNF-alpha as the inducer and mediator of neurodegeneration and their evidence suggest that TNF-alpha kill neurons directly by recruiting caspases or may kill indirectly by various means. In sharp contrast to this, another concept theory envisages a role for TNF-alpha in negotiating neuroprotection during HAD. The current compilation examines these contradictory concepts, and evaluates their efficacy in the light of TNF-alpha signaling. It also attempts to elaborate the current consensus outlook of TNF-alpha's role during HAD. PMID- 12911615 TI - Glucocorticoid-regulated human serotonin transporter (5-HTT) expression is modulated by the 5-HTT gene-promotor-linked polymorphic region. AB - Mood, emotion and cognition are modulated by serotonergic neurotransmission, while the physiological function of serotonergic synapses depends on serotonin reuptake, which is mediated by the serotonin transporter (5-HTT). Allelic variation of 5-HTT expression in humans is caused by a functional gene-promoter polymorphism with two predominant variant alleles, which are associated with variations in anxiety measures as previously reported. Here we report that administration of dexamethasone, a potent glucocorticosteroid hormone, results in an increase in 5-HTT expression in immortalized human B-lymphoblastoid cells, which express the human 5-HTT. Functional reporter gene assays as well as 5-HT uptake and inhibitor binding measures revealed a genotype-dependent dose-response to glucocorticosteroid administration, which was antagonized by RU 38486, a non specific glucocorticosteroid hormone antagonist. The allele-specific differences after administration of dexamethasone depended on the repetitive GC-rich sequence located approximately 1.4 kb upstream of the 5-HTT gene transcription site because of absence of a significant steroid effect after transfecting a deletional mutant reporter gene construct, which lacks this repetitive promoter sequence. Our findings may contribute to explain the vulnerability to stress related disorders in susceptible individuals, in whom further clinical studies should follow up on these in vitro findings. PMID- 12911616 TI - Potassium channel gene therapy can prevent neuron death resulting from necrotic and apoptotic insults. AB - Necrotic insults such as seizure are excitotoxic. Logically, membrane hyperpolarization by increasing outwardly conducting potassium channel currents should attenuate hyperexcitation and enhance neuron survival. Therefore, we overexpressed a small-conductance calcium-activated (SK2) or voltage-gated (Kv1.1) channel via viral vectors in cultured hippocampal neurons. We found that SK2 or Kv1.1 protected not only against kainate or glutamate excitotoxicity but also increased survival after sodium cyanide or staurosporine. In vivo overexpression of either channel in dentate gyrus reduced kainate-induced CA3 lesions. In hippocampal slices, the kainate-induced increase in granule cell excitability was reduced by overexpression of either channel, suggesting that these channels exert their protective effects during hyperexcitation. It is also important to understand any functional disturbances created by transgene overexpression alone. In the absence of insult, overexpression of Kv1.1, but not SK2, reduced baseline excitability in dentate gyrus granule cells. Furthermore, while no behavioral disturbances during spatial acquisition in the Morris water maze were observed with overexpression of either channel, animals overexpressing SK2, but not Kv1.1, exhibited a memory deficit post-training. This difference raises the possibility that the means by which these channel subtypes protect may differ. With further development, potassium channel vectors may be an effective pre-emptive strategy against necrotic insults. PMID- 12911617 TI - Antipsychotic drugs differentially modulate apolipoprotein D in rat brain. AB - Apolipoprotein-D (apoD), a member of the lipocalin family of proteins, binds to arachidonic acid and cholesterol among other hydrophobic molecules. Recently, elevated apoD levels have been reported in the post-mortem brains, as well as plasma, of schizophrenic patients and in rodent brains after chronic treatment with clozapine (CLOZ). These findings and the evidence for altered membrane lipid metabolism in schizophrenia suggest that apoD may have a role in the pathophysiology of illness, and also in the differential clinical outcome following treatment with typical and atypical antipsychotic drugs. Here, we compared the effects of these antipsychotics on the expression of apoD in rat brain. Chronic treatment with typical antipsychotic, haloperidol (HAL) reduced apoD expression in hippocampus, piriform cortex and caudate-putamen (p = 0.027 0.002), whereas atypical antipsychotics, risperidone (RISP) and olanzapine (OLZ) increased (p = 0.051 to < 0.001 and p = 0.048 to < 0.001, respectively) apoD expression. In hippocampus, HAL-induced changes were present in CA1, CA3 and dentate gyrus, however, apoD levels in motor cortex were unchanged. There were also very dramatic effects of HAL on the neuronal morphology, particularly, cellular shrinkage and disorganization with the loss of neuropil. Post-treatment, either with RISP or OLZ, was very effective in restoring the HAL-induced reduction of apoD, as well as cellular morphology. Similarly, pre-treatments were also effective, but slightly less than post-treatment, in preventing HAL-induced reduction of apoD. The increased expression of apoD by atypical antipsychotics may reflect a novel molecular mechanism underlying their favorable effects compared with HAL on cognition, negative symptoms and extra-pyramidal symptoms in schizophrenia. PMID- 12911618 TI - Regulation of brain mitochondrial H2O2 production by membrane potential and NAD(P)H redox state. AB - Mitochondrial production of reactive oxygen species (ROS) at Complex I of the electron transport chain is implicated in the etiology of neural cell death in acute and chronic neurodegenerative disorders. However, little is known regarding the regulation of mitochondrial ROS production by NADH-linked respiratory substrates under physiologically realistic conditions in the absence of respiratory chain inhibitors. This study used Amplex Red fluorescence measurements of H2O2 to test the hypothesis that ROS production by isolated brain mitochondria is regulated by membrane potential (DeltaPsi) and NAD(P)H redox state. DeltaPsi was monitored by following the medium concentration of the lipophilic cation tetraphenylphosphonium with a selective electrode. NAD(P)H autofluorescence was used to monitor NAD(P)H redox state. While the rate of H2O2 production was closely related to DeltaPsi and the level of NAD(P)H reduction at high values of DeltaPsi, 30% of the maximal rate of H2O2 formation was still observed in the presence of uncoupler (p-trifluoromethoxycarbonylcyanide phenylhydrazone) concentrations that provided for maximum depolarization of DeltaPsi and oxidation of NAD(P)H. Our findings indicate that ROS production by mitochondria oxidizing physiological NADH-dependent substrates is regulated by DeltaPsi and by the NAD(P)H redox state over ranges consistent with those that exist at different levels of cellular energy demand. PMID- 12911619 TI - Conditions associated with ER dysfunction activate homer 1a expression. AB - Homer proteins physically link metabotropic glutamate receptors with IP3 receptors located at the endoplasmic reticulum (ER) and thereby modulate receptor activated calcium signaling. Homer 1a, the short form of constitutively expressed homer 1 proteins, exerts dominant negative activity with respect to homer 1 proteins by interfering with the formation of multiprotein complexes. Homer 1a is an immediate early gene, the expression of which is activated by various stimuli including glutamate receptor activation. The mechanisms underlying activation of homer 1a expression are however, not fully understood. Here, we show that homer 1a expression is induced in neuronal cell cultures under experimental conditions associated with ER dysfunction. Increased homer 1a mRNA levels were found in 2 sets of cultures: in those exposed to thapsigargin, a specific inhibitor of ER Ca2+-ATPase, after a transient depletion of ER calcium stores through exposure to calcium-free medium supplemented with EGTA, and in those exposed to a proteasome inhibitor known to induce ER dysfunction. Thus, homer 1a expression may be activated by impairment of ER functioning just as it is by glutamate receptor activation. PMID- 12911620 TI - The synergistic effects of NGF and IGF-1 on neurite growth in adult sensory neurons: convergence on the PI 3-kinase signaling pathway. AB - Nerve growth factor (NGF) and insulin-like growth factor-1 (IGF-1) play an important role in promoting axonal growth from dorsal root ganglion (DRG) neurons. Adult DRG neurons exhibit neurotrophin-independent survival, providing an excellent system with which to study trophic factor effects on neurite growth in the absence of significant survival effects. Using young adult rat DRG neurons we have demonstrated a synergistic effect of NGF plus IGF (N + I), compared with either factor alone, in promoting neurite growth. Not only does the presence of NGF and IGF-1 enhance neurite initiation, it also significantly augments the extent of neurite branching and elongation. We have also examined potential mechanism(s) underlying this synergistic effect. Immunoblotting experiments of classical growth factor intermediary signalling pathways (PI 3-K-Akt-GSK-3 and Ras-Raf-MAPK) were performed using phospho-specific antibodies to assess activation state. We found that activation of Akt and MAPK correlated with neurite elongation and branching. However, using pharmacological inhibitors, we observed that a PI 3-K pathway involving both Akt and GSK-3 appeared to be more important for neurite extension and branching than MAPK-dependent signalling. In fact, inhibition of activation of MAPK with U0126 resulted in increased neuritic branching, possibly as a result of the concomitant increase observed in phospho Akt. Furthermore, inhibition of GSK3 (which is negatively regulated by phosphorylation on S9/S21) also resulted in increased growth. Our data point to signalling convergence upon the PI 3-K-Akt-GSK-3 pathway that underlies the NGF plus IGF synergism. In addition, to our knowledge, this is the first report in primary neurons that inhibition of GSK3 results in an enhanced neurite growth. PMID- 12911621 TI - Systemic morphine-induced release of serotonin in the rostroventral medulla is not mimicked by morphine microinjection into the periaqueductal gray. AB - We used in vivo microdialysis in awake rats to test the hypothesis that intravenous morphine increases serotonin (5-HT) release within the rostral ventromedial medulla (RVM). We also injected morphine into various sites along the rostrocaudal extent of the periaqueductal gray (PAG), and examined the extent of its diffusion to the RVM. Intravenous morphine (3.0 mg/kg) produced thermal antinociception and increased RVM dialysate 5-HT, 5-hydroxyindole acetic acid (5 HIAA), and homovanillic acid (HVA) in a naloxone-reversible manner. As neither PAG microinjection of morphine (5 micro g/0.5 micro L) nor RVM administration of fentanyl or d-Ala(2),NMePhe(4),Gly-ol(5)]enkephalin (DAMGO) increased RVM 5-HT, we were unable to determine the precise site of action of morphine. Surprisingly, peak morphine levels in the RVM were higher after microinjection into the caudal PAG as compared to either intravenous injection or microinjection into more rostral sites within the PAG. Naloxone-precipitated withdrawal in morphine tolerant rats not only increased extracellular 5-HT in the RVM, but also dopamine (DA) and HVA. We conclude that substantial amounts of morphine diffuse from the PAG to the RVM, and speculate that opioid receptor interactions at multiple brain sites mediate the analgesic effects of PAG morphine. Further studies will be required to elucidate the contribution of 5-HT and DA release in the RVM to opioid analgesia and opioid withdrawal. PMID- 12911622 TI - Neuromelanin associated redox-active iron is increased in the substantia nigra of patients with Parkinson's disease. AB - Degeneration of dopaminergic neurones during Parkinson's disease is most extensive in the subpopulation of melanized-neurones located in the substantia nigra pars compacta. Neuromelanin is a dark pigment produced in the dopaminergic neurones of the human substantia nigra and has the ability to bind a variety of metal ions, especially iron. Post-mortem analyses of the human brain have established that oxidative stress and iron content are enhanced in association with neuronal death. As redox-active iron (free Fe2+ form) and other transition metals have the ability to generate highly reactive hydroxyl radicals by a catalytic process, we investigated the redox activity of neuromelanin (NM) aggregates in a group of parkinsonian patients, who presented a statistically significant reduction (- 70%) in the number of melanized-neurones and an increased non-heme (Fe3+) iron content as compared with a group of matched control subjects. The level of redox activity detected in neuromelanin-aggregates was significantly increased (+ 69%) in parkinsonian patients and was highest in patients with the most severe neuronal loss. This change was not observed in tissue in the immediate vicinity of melanized-neurones. A possible consequence of an overloading of neuromelanin with redox-active elements is an increased contribution to oxidative stress and intraneuronal damage in patients with Parkinson's disease. PMID- 12911623 TI - A new G protein-coupled receptor from a primitive metazoan shows homology with vertebrate aminergic receptors and displays constitutive activity in mammalian cells. AB - Biogenic amine receptors mediate wide-ranging hormonal and modulatory functions in vertebrates, but are largely unknown in primitive invertebrates. In a representative of the most basal multicellular animals possessing a nervous system, the cnidarian Renilla koellikeri, aminergic-like receptors were previously characterized pharmacologically and found to engender control of the animal's bioluminescent and peristaltic reactions. Using degenerate oligonucleotides in a RT-PCR strategy, we obtained a full-length cDNA encoding a polypeptide with typical G protein-coupled receptor (GPCR) characteristics and which displayed a significant degree of sequence similarity (up to 45%) to biogenic amine receptors, particularly dopamine and adrenergic receptors. The new receptor, named Ren1, did not resemble any one specific type of amine GPCR and thus could not be identified on the basis of sequence. Ren1 was expressed transiently and stably in cultured mammalian cells, as demonstrated by immunocytochemistry and western blotting. Functional analysis of transfected HEK293, LTK- and COS-7 cells, based on both cAMP and Ca2+ signalling assays, revealed that Ren1 was not activated by any of the known biogenic amines tested and several related metabolites. The results indicated, however, that cells stably expressing Ren1 contained, on average, an 11-fold higher level of cAMP than the controls, in the absence of agonist stimulation. The high basal cAMP levels were shown to be specific for Ren1 and to vary proportionally with the level of Ren1 expressed in the transfected cells. Taken together, the data suggested that Ren1 was expressed as a constitutively active receptor. Its identification provides a basis for examination of the early evolutionary emergence of GPCRs and their functional properties. PMID- 12911624 TI - Activity of the rat GluR4 promoter in transfected cortical neurons and glia. AB - AMPA (alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate) receptors are assembled from four subunits, GluR1-4. Although GluR4 is widely expressed in brain its abundance is less than GluR1-3. We have isolated approximately 5 kb of the rat GluR4 promoter region and analyzed its capacity to drive expression of a luciferase reporter gene in transfected rat cortical neurons and glia, and C6 glioma cells. Multiple transcriptional start sites were identified in a GC-rich region lacking TATA-boxes between -1090 and -1011 bp from ATG. In transfected mixed cortical cultures, luciferase expression driven by GluR4 promoter segments were found predominantly in TuJ1-positive neurons, indicating neuronal preference of GluR4. The GluR4 promoter fragments were 6-12-fold more active in neurons than glia, compared with a 30-fold neuronal selectivity of GluR2. Deletion of the GluR4 transcriptional initiation region decreased luciferase activity in neurons, but increased activity in C6 cells, suggesting that regulatory elements governing neuronal expression reside in this region. An intron within the 5'-untranslated region and Sp1, IK2 and E-box sites are conserved in the rat, mouse and human GluR4 promoters. The relative activity of GluR4 and GluR2 promoters in transfected cells correlates with their expression in brain, and in both promoters regulatory elements for neuronal expression reside near the initiation sites. PMID- 12911625 TI - Cytochrome P450-dependent metabolism of l-deprenyl in monkey (Cercopithecus aethiops) and C57BL/6 mouse brain microsomal preparations. AB - The aim of the present investigation was to characterize the cytochrome P450 (CYP)-dependent metabolism of l-deprenyl by brain microsomal preparations obtained from two different animal models that have been extensively used in Parkinson's disease studies, namely monkey (Cercopithecus aethiops) and C57BL/6 mouse. In monkey brain microsomal fractions, the apparent Km values for methamphetamine formation from l-deprenyl were 67.8 +/- 1.0 and 72.0 +/- 1.6 microm, in the cortex and striatum, respectively. Similarly, for nordeprenyl formation from l-deprenyl, Km values in cortex and striatum were 21.3 +/- 3.2 and 27.3 +/- 4.0 microm, respectively. Both metabolic pathways appear to be more efficient in the cortex than in the striatum as the Vmax for microsomal preparation was lower in the striatum for the formation of both metabolites. The formation rate of l-methamphetamine was up to one order of magnitude greater than that of nordeprenyl. Inhibition analysis of both pathways in monkey brain suggested that l-methamphetamine formation is catalysed by CYP2A and CYP3A, whereas only CYP3A appears to be involved in nordeprenyl formation. With microsomal preparations from whole brain of C57BL/6 mice, the only l-deprenyl metabolite that could be detected was methamphetamine and the Km and Vmax values were similar to those determined in monkey cortex (53.6 +/- 2.9 microm and 33.9 +/- 0.4 pmol/min/mg protein, respectively). 4-Methylpyrazole selectively inhibited methamphetamine formation, suggesting the involvement of CYP2E1. In conclusion, the present study indicates that l-deprenyl is effectively metabolised by CYP-dependent oxidases in the brain, giving rise mainly to the formation of methamphetamine, which has been suggested to play a role in the pharmacological effects of the parent drug. The results also demonstrate that there are differences between species in CYP-dependent metabolism of l-deprenyl. PMID- 12911626 TI - Regulation of the glutamate transporter EAAT1 by the ubiquitin ligase Nedd4-2 and the serum and glucocorticoid-inducible kinase isoforms SGK1/3 and protein kinase B. AB - Surface expression of the glial glutamate transporter EAAT1 is stimulated by insulin-like growth factor 1 through activation of phosphatidylinositol-3-kinase. Downstream targets include serum and glucocorticoid-sensitive kinase isoforms SGK1, SGK2 and SGK3, and protein kinase B. SGK1 regulates Nedd4-2, a ubiquitin ligase that prepares cell membrane proteins for degradation. To test whether Nedd4-2, SGK1, SGK3 and protein kinase B regulate EAAT1, cRNA encoding EAAT1 was injected into Xenopus oocytes with or without additional injection of wild-type Nedd4-2, constitutively active S422DSGK1, inactive K127NSGK1, wild-type SGK3 and/or constitutively active T308D,S473DPKB. Glutamate induces a current in Xenopus oocytes expressing EAAT1, but not in water-injected oocytes, which is decreased by co-expression of Nedd4-2, an effect reversed by additional co expression of S422DSGK1, SGK3 and T308D,S473DPKB, but not K127NSGK1. Site directed mutagenesis of the SGK1 phosphorylation sites in the Nedd4-2 protein (S382A,S468ANedd4-2) and in the EAAT1 protein (T482AEAAT1, T482DEAAT1) significantly blunts the effect of S422DSGK1. Moreover, the current is significantly larger in T482DEAAT1- than in T482AEAAT1-expressing oocytes, indicating that a negative charge mimicking phosphorylation at T482 increases transport. The experiments reveal a powerful novel mechanism that regulates the activity of EAAT1. This mechanism might participate in the regulation of neuronal excitability and glutamate transport in other tissues. PMID- 12911627 TI - Green tea polyphenols enhance sodium nitroprusside-induced neurotoxicity in human neuroblastoma SH-SY5Y cells. AB - Oxidative stress is a main mediator in nitric oxide (NO) -induced neurotoxicity and has been implicated in the pathogenesis of many neurodegenerative disorders. Green tea polyphenols are usually expected as potent chemo-preventive agents due to their ability of scavenging free radicals and chelating metal ions. However, not all the actions of green tea polyphenols are necessarily beneficial. In the present study, we demonstrated that higher-concentration green tea ployphenols significantly enhanced the neurotoxicity by treatment of sodium nitroprusside (SNP), a nitric oxide donor. SNP induced apoptosis in human neuroblastoma SH-SY5Y cells in a concentration and time-dependent manner, as estimated by cell viability assessment, FACScan analysis and DNA fragmentation assay, whereas treatment with green tea polyphenols alone had no effect on cell viability. Pre treatment with lower-dose green tea polyphenols (50 and 100 microm) had only a slightly deleterious effect in the presence of SNP, while higher-dose green tea polyphenols (200 and 500 microm) synergistically damaged the cells severely. Further research showed that co-incubation of green tea polyphenols and SNP caused loss of mitochondrial membrane potential, depletion of intracellular GSH and accumulation of reactive oxygen species, and exacerbated NO-induced neuronal apoptosis via a Bcl-2 sensitive pathway. PMID- 12911628 TI - Lipopolysaccharide-induced dopaminergic cell death in rat midbrain slice cultures: role of inducible nitric oxide synthase and protection by indomethacin. AB - Glial cell activation associated with inflammatory reaction may contribute to pathogenic processes of neurodegenerative disorders, through production of several cytotoxic molecules. We investigated the consequences of glial activation by interferon-gamma (IFN-gamma)/lipopolysaccharide (LPS) in rat midbrain slice cultures. Application of IFN-gamma followed by LPS caused dopaminergic cell death and accompanying increases in nitrite production and lactate dehydrogenase release. Aminoguanidine, an inhibitor of inducible nitric oxide synthase (iNOS), or SB203580, an inhibitor of p38 mitogen-activated protein kinase, prevented dopaminergic cell loss as well as nitrite production. SB203580 also suppressed expression of iNOS and cyclooxygenase-2 (COX-2) induced by IFN-gamma/LPS. A COX inhibitor indomethacin protected dopaminergic neurons from IFN-gamma/LPS-induced injury, whereas selective COX-2 inhibitors such as NS-398 and nimesulide did not. Notably, indomethacin was able to attenuate neurotoxicity of a nitric oxide (NO) donor. Neutralizing antibodies against tumour necrosis factor-alpha and interleukin-1beta did not inhibit dopaminergic cell death caused by IFN gamma/LPS, although combined application of these antibodies blocked lactate dehydrogenase release and decrease in the number of non-dopaminergic neurons. These results indicate that iNOS-derived NO plays a crucial role in IFN-gamma/LPS induced dopaminergic cell death, and that indomethacin exerts protective effect by mechanisms probably related to NO neurotoxicity rather than through COX inhibition. PMID- 12911629 TI - A novel function of Goalpha: mediation of extracellular signal-regulated kinase activation by opioid receptors in neural cells. AB - Go is the most abundant G protein expressed in brain but its function is less known. Here we show a novel function of Goalpha as a mediator of opioid receptor induced extracellular signal-regulated kinase activation in neural cells. The current study found that, in neuroblastoma x glioma NG108-15 hybrid cells, activation of extracellular signal-regulated kinase through delta opioid receptors was mediated by pertussis toxin-sensitive G protein and independent of Gbetagamma subunits, PI3 kinase and receptor internalization. Overexpression of a dominant negative form of Goalpha1, but not Gialpha2, completely blocked delta opioid receptor-induced extracellular signal-regulated kinase activity. Decreasing Goalpha expression by RNA interference greatly reduced delta opioid receptor-induced extracellular signal-regulated kinase activity and extracellular signal-regulated kinase-dependent gene expression, while knocking down Gialpha2 did not. By taking advantage of differences between human and mouse Goalpha gene sequences, we simultaneously knocked down endogenous Goalpha expression and expressed exogenous human Goalpha subunits. We found that both human Goalpha1 and Goalpha2 could mediate delta opioid receptor-induced extracellular signal regulated kinase activation. This study suggests that one of the functions of Goalpha in the brain is to mediate extracellular signal-regulated kinase activation by G protein-coupled receptors. PMID- 12911630 TI - Both apoptosis and necrosis occur early after intracerebral grafting of ventral mesencephalic tissue: a role for protease activation. AB - Neural transplantation is an experimental treatment for Parkinson's disease. Widespread clinical application of the grafting technique is hampered by a relatively poor survival (around 10%) of implanted embryonic dopamine neurones. Earlier animal studies have indicated that a large proportion of the grafted cells die during graft tissue preparation and within the first few days after intracerebral implantation. The present study was designed to reveal the prevalence of cell death in rat intrastriatal grafts at 90 min, 1, 3, 6 and 42 days after implantation. We examined apoptotic cell death using semi-thin and paraffin sections stained with methylene blue and an antibody against activated caspase 3, respectively. We identified abundant apoptotic cell death up to 3 days after transplantation. In addition, we studied calpain activation using an antibody specific for calpain-cleaved fodrin. We report a peak in calpain activity 90 min after grafting. Surprisingly, we did not observe any significant difference in the number of dopaminergic neurones over time. The present results imply that grafted cells may be victims of either an early necrotic or a later apoptotic cell death and that there is substantial cell death as early as 90 min after implantation. PMID- 12911631 TI - Cellular distribution and bioactivity of the key steroidogenic enzyme, cytochrome P450side chain cleavage, in sensory neural pathways. AB - Neurosteroids are steroids produced within the nervous system. Based on behavioural responses evoked in animals by synthetic steroid injections, several studies suggested neurosteroid involvement in important neurophysiological processes. These observations should be correlated only to neuroactive effects of the injected steroids. Neurosteroids mostly control the CNS activity through allosteric modulation of neurotransmitter receptors within concentration ranges used by neurotransmitters themselves. Therefore, neurosteroid production within pathways controlling a neurophysiological process is necessary to consider neurosteroid involvement in that process. Because of the increasing speculation about pain modulation by neurosteroids based on pharmacological observations, we decided to clarify the situation by investigating neurosteroidogenesis occurrence in sensory pathways, particularly in nociceptive structures. We studied the presence and activity of cytochrome P450side chain cleavage (P450scc) in rat pain pathways. P450scc-immunoreactive cells were localized in dorsal root ganglia (DRG), spinal cord (SC) dorsal horn, nociceptive supraspinal nuclei (SSN) and somatosensory cortex. Incubation of DRG, SSN or SC tissue homogenates with [3H]cholesterol yielded the formation of radioactive metabolites including [3H]pregnenolone of which the synthesis was reduced in presence of aminogluthetimide, a P450scc inhibitor. These first neuroanatomical and neurochemical results demonstrate the occurrence of neurosteroidogenesis in nociceptive pathways and strongly suggest that neurosteroids may control pain mechanisms. PMID- 12911632 TI - Hydrogen peroxide induces loss of dopamine transporter activity: a calcium dependent oxidative mechanism. AB - H2O2 dose dependently inhibited dopamine uptake in PC12 cells and in striatal synaptosomes. Treatment with H2O2 resulted in a reversible reduction in Vmax, with no effect on its Km value. This suppressive effect of H2O2 could be relieved by reducing agents (dithiothreitol and cysteine). Furthermore, an oxidizer (dithiodipyridine) also markedly suppressed the dopamine transporter (DAT). Oxidative stress therefore might contribute to the action of H2O2. H2O2 appeared to modify DAT at both extracellular and intracellular sites because cumene-H2O2 (a radical generator mostly restricted to plasma membranes) at high concentrations also slightly suppressed DAT activity and the intracellular overexpression of catalase ameliorated the inhibitory effect of H2O2. Internalization was unlikely to be involved because concanavalin A, which blocked endocytosis, did not prevent the H2O2-evoked inhibition of DAT activity. Interestingly, H2O2 treatment evoked a Ca2+ influx in PC12 cells. Moreover, removal of external calcium by EGTA or reduction in the intracellular calcium level using BAPTA-AM reversed the inhibitory effect of H2O2. Conversely, depletion of intracellular calcium stores using thapsigargin did not affect the reduction in DAT activity by H2O2. Collectively, our results indicate that the DAT, one of the most important proteins controlling the dopaminergic system, is also a redox sensor. In addition, H2O2 might suppress the DAT by a Ca2+-dependent oxidative pathway. PMID- 12911633 TI - Expression of death-associated protein kinase and recruitment to the tumor necrosis factor signaling pathway following brief seizures. AB - Death-associated protein (DAP) kinase is calcium-regulated and known to function downstream of death receptors, prompting us to examine its role in the mechanism of seizure-induced neuronal death. Brief seizures were focally evoked in rats, eliciting neuronal death within the CA3 subfield of the hippocampus, and to a lesser extent, cortex. Western blotting confirmed expression of DAP kinase within hippocampus and cortex at the predicted weight of approximately 160 kDa. Immunohistochemistry revealed seizures triggered a significant increase in numbers of DAP kinase-expressing cells within CA3 and cortex, without affecting cell counts within seizure-resistant CA2 or the dentate gyrus. Numbers of DAP kinase-expressing cells were increased in relation to specific patterns of injury causing seizure activity, electrographically defined. Seizures caused an early increase in DAP kinase binding to actin, and association with calmodulin. Co immunoprecipitation studies also revealed seizures triggered binding of DAP kinase to the tumor necrosis factor receptor 1 and the Fas-associated death domain protein, commensurate with caspase-8 proteolysis. In contrast, within surviving fields of the hippocampus, DAP kinase interacted with the molecular chaperone 14-3-3. These data suggest DAP kinase is involved in the molecular pathways activated during seizure-induced neuronal death. PMID- 12911634 TI - UDP-N-acetylglucosaminyl transferase (OGT) in brain tissue: temperature sensitivity and subcellular distribution of cytosolic and nuclear enzyme. AB - In brain tissue, UDP-N-acetylglucosaminyl transferase (OGT) is known to catalyze the addition of a single N-acetylglucosamine moiety (GlcNAc) onto two proteins linked to the etiology of neurodegenerative disease--beta-amyloid associated protein and tau. Hyperphosphorylation of tau appears to cause neurofibrillary tangles and cell death, and a functional relationship appears to exist between phosphorylation and glycosylation. Since a greater understanding of brain OGT may provide new insights into the pathogenesis of Alzheimer's disease, we examined the characteristics and subcellular distribution of OGT protein and OGT activity and its relationship to O-linked glycosylation. We found that cytosolic OGT activity is 10 times more abundant in brain tissue compared with muscle, adipose, heart, and liver tissue. Temperature studies demonstrated that cytosolic OGT activity was stable at 24 degrees C but was rapidly inactivated at 37 degrees C (T1/2 = 20 min). Proteases were probably not involved because OGT immunopurified from cytosol retained temperature sensitivity. Subcellular distribution studies showed abundant OGT protein in the nucleus that was enzymatically active. Nuclear OGT activity exhibited a high affinity for UDP-GlcNAc and a salt sensitivity that was similar to cytosolic OGT; however, nuclear OGT was not inactivated at 37 degrees C, as was the cytosolic enzyme. Two methods were used to measure O-linked glycoproteins in brain cytosol and nucleosol -[3H]galactose labeling and western blotting using antibodies against O-linked glycoproteins. Both methods revealed a greater abundance of O-linked glycoproteins in the nucleus compared to cytosol. PMID- 12911635 TI - Erasure of kinase phosphorylation in astrocytes during oxygen-glucose deprivation is controlled by ATP levels and activation of phosphatases. AB - We have examined the relationship between adenosine triphosphate (ATP) concentration and loss of maintenance of kinase-signalling cascades in primary cortical astrocytes during oxygen-glucose deprivation (OGD) as this may constitute an irreversible step that commits astrocytes to cell death. We report that the phosphorylation of Akt, ERK, JNK and p38 kinases, whose activities depend on serine, threonine and tyrosine phosphorylation, were all increased during OGD. All these phosphorylations were reduced to below detection limits when ATP levels were less than 10% of normal levels. Using ERK and Akt as representative examples, we show that this erasure is not irreversible as both ERK and Akt phosphorylations can be partially restored by addition of glucose under anoxic conditions. We further investigated whether OGD caused any change in phosphatase activity. The PP1/PP2A phosphatase inhibitors okadaic acid and caliculyn A, but not cyclosporine A, delayed the removal of ERK and Akt phosphorylation under OGD. By comparing the extent of phosphorylation increase under OGD and normoxic conditions, we calculate that phosphatase activity was increased by approximately 3.6-fold during OGD. These data show that ATP levels control an important checkpoint in kinase function, and that ATP levels may need to be considered when studies of kinase function in relation to OGD are conducted. PMID- 12911636 TI - Identification of members of the Bex gene family as olfactory marker protein (OMP) binding partners. AB - Olfactory marker protein (OMP) expression is a hallmark of mature vertebrate olfactory receptor neurons (ORNs). Evidence for OMP function derives from altered behavioral and electrophysiological activities of OMP-KO mice. The molecular basis for the altered phenotype following the deletion of OMP is still unclear. Recent structural studies predict the involvement of OMP in protein-protein interaction. Here we report the identification of an OMP partner, Bex2, by phage display screening of an olfactory mucosal cDNA-library. In situ hybridization demonstrates cellular co-localization of OMP mRNA with mRNAs for Bex1, Bex2, and Bex3 in ORNs of olfactory tissue of the mouse. The OMP/Bex interaction has been confirmed by demonstrating the chemical cross-linking of recombinant rat OMP with a synthetic peptide derived from the Bex amino acid sequence. The subcellular localization of Bex and OMP proteins was evaluated in transfected HEK293 cells. Bex is visualized in the nucleus and cytoplasm. Following co-transfection we observed the unexpected presence of some OMP in the nucleus along with Bex. Together, these data argue convincingly that we have identified Bex as an OMP partner whose further characterization will provide insight to the role of OMP and to the mechanism of the OMP/Bex interaction in ORN differentiation and function. PMID- 12911637 TI - Dysfunction of mitochondrial complex I and the proteasome: interactions between two biochemical deficits in a cellular model of Parkinson's disease. AB - Two biochemical deficits have been described in the substantia nigra in Parkinson's disease, decreased activity of mitochondrial complex I and reduced proteasomal activity. We analysed interactions between these deficits in primary mesencephalic cultures. Proteasome inhibitors (epoxomicin, MG132) exacerbated the toxicity of complex I inhibitors [rotenone, 1-methyl-4-phenylpyridinium (MPP+)] and of the toxic dopamine analogue 6-hydroxydopamine, but not of inhibitors of mitochondrial complex II-V or excitotoxins [N-methyl-d-aspartate (NMDA), kainate]. Rotenone and MPP+ increased free radicals and reduced proteasomal activity via adenosine triphosphate (ATP) depletion. 6-hydroxydopamine also increased free radicals, but did not affect ATP levels and increased proteasomal activity, presumably in response to oxidative damage. Proteasome inhibition potentiated the toxicity of rotenone, MPP+ and 6-hydroxydopamine at concentrations at which they increased free radical levels >/= 40% above baseline, exceeding the cellular capacity to detoxify oxidized proteins reduced by proteasome inhibition, and also exacerbated ATP depletion caused by complex I inhibition. Consistently, both free radical scavenging and stimulation of ATP production by glucose supplementation protected against the synergistic toxicity. In summary, proteasome inhibition increases neuronal vulnerability to normally subtoxic levels of free radicals and amplifies energy depletion following complex I inhibition. PMID- 12911638 TI - Comparison of circadian expression of tryptophan hydroxylase isoform mRNAs in the rat pineal gland using real-time PCR. AB - A second gene encoding a functional tryptophan hydroxylase activity has recently been described (TPH2), which is expressed abundantly in brainstem, the primary site of serotonergic neurons in the CNS. As serotonin (5-HT) has an important role as a precursor of the nocturnal synthesis of the pineal gland hormone, melatonin, it was of interest to determine the relative expression of TPH1 and 2 mRNA in the rat pineal during the light:dark (L:D) cycle using sensitive real time RT-PCR assays which were developed for each TPH isoform. TPH1 mRNA expression was 105-fold more abundant in rat pineal than TPH2, and showed a significant approximately 4-fold nocturnal increase in expression which may contribute to the previously described nocturnal increase in pineal tryptophan hydroxylase activity. TPH2 expression within the gland showed no significant variation with time of day and was very low (approximately 300 copies/gland) indicating expression in the small proportion of "non-pinealocyte" cells in the gland. PMID- 12911640 TI - Detection of errors by attending physicians on a general medicine service. AB - BACKGROUND: Attending physicians are well positioned to identify medical errors and understand their consequences. The spectrum of errors that can be detected by attending physicians in the course of their usual practice is currently unknown. OBJECTIVES: To determine the frequency, types, and consequences of errors that can be detected by attending hospitalist physicians in the care of their patients, and to compare the types of errors first discovered by attending hospitalists to those discovered by other providers. DESIGN: Prospective identification of errors by attending physicians. SETTING: Two hundred-bed, academic hospital. PATIENTS: Five hundred twenty-eight patients admitted to the general medicine service from October 2000 to April 2001. MEASUREMENTS: Errors, both near misses and adverse events, were identified during the course of routine, clinical care by 2 attending hospitalists. Errors first detected by other health care workers were also recorded. MAIN RESULTS: Of the 528 patients admitted to the hospitalist service, 10.4% experienced at least 1 error: 6.2% a near miss and 4.2% an adverse event. Although differences did not achieve statistical significance, most of the errors first detected by house staff, nurses, and laboratory technicians were adverse events; most of the errors first detected by the attending hospitalists, pharmacists, and consultants were near misses. Drug errors were the most common type of error overall. CONCLUSIONS: Attending physicians engaged in routine clinical care can detect a range of errors, and differences may exist in the types of errors detected by various health care providers. PMID- 12911641 TI - Mortality and length of stay in a veterans affairs hospital and private sector hospitals serving a common market. AB - OBJECTIVE: To compare severity-adjusted in-hospital mortality and length of stay (LOS) in a Veterans Administration (VA) hospital and private sector hospitals serving the same health care market. DESIGN: Retrospective cohort study. SETTING: A large VA hospital and 27 private sector hospitals in the same metropolitan area. PATIENTS: Consecutive VA (N = 1,960) and private sector (N = 157,147) admissions in 1994 to 1995 with 9 high-volume diagnoses. MEASUREMENTS: Severity of illness was measured using validated multivariable models that were based on data abstracted from medical records. Outcomes were adjusted for severity and compared in VA and private sector patients using multiple logistic or linear regression analysis. MAIN RESULTS: Unadjusted mortality was similar in VA and private sector patients (5.0% vs 5.6%, respectively; P =.26), although mean LOS was longer in VA patients (12.7 vs 7.0 days; P <.001). Adjusting for severity, the odds of death in VA patients was similar (odds ratio [OR] 1.07; 95% confidence interval [95% CI], 0.74 to 1.54; P =.73). However, a larger proportion of deaths in VA patients occurred later during hospitalization (P <.001), and the odds of death in VA patients were actually lower (P <.05) in analyses limited to deaths during the first 7 (OR, 0.56) or 14 (OR, 0.63) days. Adjusted LOS was longer (P <.001) in VA patients for all 9 diagnoses. CONCLUSIONS: If the current findings generalizable to other markets, hospital mortality, a widely used performance measure, may be similar or lower in VA and private sector hospitals serving the same markets. The longer LOS of VA patients may reflect differences in practice patterns and may be an important source of bias in comparisons of VA and private sector hospitals. PMID- 12911642 TI - Beliefs about control in the physician-patient relationship: effect on communication in medical encounters. AB - OBJECTIVES: Effective communication is a critical component of quality health care, and to improve it we must understand its dynamics. This investigation examined the extent to which physicians' and patients' preferences for control in their relationship (e.g., shared control vs doctor control) were related to their communications styles and adaptations (i.e., how they responded to the communication of the other participant). DESIGN: Stratified case-controlled study. PATIENTS/PARTICIPANTS: Twenty family medicine and internal medicine physicians and 135 patients. MEASUREMENTS: Based on scores from the Patient Practitioner Orientation Scale, 10 patient-centered physicians (5 male, 5 female) and 10 doctor-centered physicians (5 male, 5 female) each interacted with 5 to 8 patients, roughly half of whom preferred shared control and the other half of whom were oriented toward doctor control. Audiotapes of 135 consultations were coded for behaviors indicative of physician partnership building and active patient participation. MAIN RESULTS: Patients who preferred shared control were more active participants (i.e., expressed more opinions, concerns, and questions) than were patients oriented toward doctor control. Physicians' beliefs about control were not related to their use of partnership building. However, physicians did use more partnership building with male patients. Not only were active patient participation and physician partnership building mutually predictive of each other, but also approximately 14% of patient participation was prompted by physician partnership building and 33% of physician partnership building was in response to active patient participation. CONCLUSIONS: Communication in medical encounters is influenced by the physician's and patient's beliefs about control in their relationship as well as by one another's behavior. The relationship between physicians' partnership building and active patient participation is one of mutual influence such that increases in one often lead to increases in the other. PMID- 12911643 TI - Survivors of intimate partner violence speak out: trust in the patient-provider relationship. AB - OBJECTIVE: To identify characteristics that facilitate trust in the patient provider relationship among survivors of intimate partner violence (IPV). DESIGN: Semistructured, open-ended interviews were conducted to elicit participants' beliefs and attitudes about trust in interactions with health care providers. Using grounded theory methods, the transcripts were analyzed for common themes. A community advisory group, composed of advocates, counselors and IPV survivors, helped interpret themes and interview excerpts. Together, key components of trust were identified. SETTING: Eastern Massachusetts. PARTICIPANTS: Twenty-seven female survivors of IPV recruited from community-based IPV organizations. MAIN RESULTS: Participants' ages ranged from 18 to 56 years, 36% were African American, 32% Hispanic, and 18% white. We identified 5 dimensions of provider behavior that were uniquely important to the development of trust for these IPV survivors: 1) communication about abuse: provider was willing to openly discuss abuse; 2) professional competency: provider asked about abuse when appropriate and was familiar with medical and social histories; 3) practice style: provider was consistently accessible, respected confidentiality, and shared decision making; 4) caring: provider demonstrated personal concern beyond biomedical role through nonjudgmental and compassionate gestures, empowering statements, and persistent, committed behaviors; 5) emotional equality: provider shared personal information and feelings and was perceived by the participant as a friend. CONCLUSIONS: These IPV survivors identified dimensions of provider behavior that facilitate trust in their clinical relationship. Strengthening these provider behaviors may increase trust with patients and thus improve disclosure of and referral for IPV. PMID- 12911644 TI - Dimensions of patient-provider communication and diabetes self-care in an ethnically diverse population. AB - BACKGROUND: Patient-provider communication is essential for effective care of diabetes and other chronic illnesses. However, the relative impact of general versus disease-specific communication on self-management is poorly understood, as are the determinants of these 2 communication dimensions. DESIGN: Cross-sectional survey. SETTING: Three VA heath care systems, 1 county health care system, and 1 university-based health care system. PATIENTS: Seven hundred fifty-two diabetes patients were enrolled. Fifty-two percent were nonwhite, 18% had less than a high school education, and 8% were primarily Spanish-speaking. MEASUREMENTS AND MAIN RESULTS: Patients' assessments of providers' general and diabetes-specific communication were measured using validated scales. Self-reported foot care; and adherence to hypoglycemic medications, dietary recommendations, and exercise were measured using standard items. General and diabetes-specific communication reports were only moderately correlated (r =.35) and had differing predictors. In multivariate probit analyses, both dimensions of communication were independently associated with self-care in each of the 4 areas examined. Sociodemographically vulnerable patients (racial and language minorities and those with less education) reported communication that was as good or better than that reported by other patients. Patients receiving most of their diabetes care from their primary provider and patients with a longer primary care relationship reported better general communication. VA and county clinic patients reported better diabetes-specific communication than did university clinic patients. CONCLUSIONS: General and diabetes-specific communication are related but unique facets of patient-provider interactions, and improving either one may improve self management. Providers in these sites are communicating successfully with vulnerable patients. These findings reinforce the potential importance of continuity and differences among VA, county, and university health care systems as determinants of patient-provider communication. PMID- 12911645 TI - Impact of interpretation method on clinic visit length. AB - OBJECTIVE: To determine the impact of interpretation method on outpatient visit length. DESIGN: Time-motion study. SETTING: Hospital-based outpatient teaching clinic. PARTICIPANTS: Patients presenting for scheduled outpatient visits. MEASUREMENTS AND MAIN RESULTS: Over a 6-week study period, a research assistant recorded the following information for consecutive patient visits: patient age, gender and insurance type; type of interpreter used (none, hospital interpreter, telephone interpreter or patient-supplied interpreter); scheduled visit length; provider type (nurse practitioner; attending physician; resident in postgraduate year 1, 2 or 3, or medical student); provider gender; amount of time the patient spent in the examination room with the provider (provider time); and total time the patient spent in the clinic from check-in to checkout (clinic time). When compared to patients not requiring an interpreter, patients using some form of interpreter had longer mean provider times (32.4 minutes [min] vs 28.0 min, P <.001) and clinic times (93.6 min vs 82.4 min, P =.002). Compared to patients not requiring an interpreter, patients using a telephone interpreter had significantly longer mean provider times (36.3 min vs 28.0 min, P <.001) and clinic times (99.9 min vs 82.4 min, P =.02). Similarly, patients using a patient supplied interpreter had longer mean provider times (34.4 min vs 28.0 min, P <.001) and mean clinic times (92.8 min vs 82.4 min, P =.027). In contrast, patients using a hospital interpreter did not have significantly different mean provider times (26.8 min vs 28.0 min, P =.51) or mean clinic times (91.0 min vs 82.4 min, P =.16) than patients not requiring an interpreter. CONCLUSION: In our setting, telephone and patient-supplied interpreters were associated with longer visit times, but full-time hospital interpreters were not. PMID- 12911646 TI - Is there a July phenomenon? The effect of July admission on intensive care mortality and length of stay in teaching hospitals. AB - BACKGROUND: It has been suggested that inexperience of new housestaff early in an academic year may worsen patient outcomes. Yet, few studies have evaluated the "July Phenomenon," and no studies have investigated its effect in intensive care patients, a group that may be particularly susceptible to deficiencies in management stemming from housestaff inexperience. OBJECTIVE: Compare hospital mortality and length of stay (LOS) in intensive care unit (ICU) admissions from July to September to admissions during other months, and compare that relationship in teaching and nonteaching hospitals, and in surgical and nonsurgical patients. DESIGN, SETTING, AND PATIENTS: Retrospective cohort analysis of 156,136 consecutive eligible patients admitted to 38 ICUs in 28 hospitals in Northeast Ohio from 1991 to 1997. RESULTS: Adjusting for admission severity of illness using the APACHE III methodology, the odds of death was similar for admissions from July through September, relative to the mean for all months, in major (odds ratio [OR], 0.96; 95% confidence interval [95% CI], 0.91 to 1.02; P =.18), minor (OR, 1.02; 95% CI, 0.93 to 1.10; P =.66), and nonteaching hospitals (OR, 0.96; 95% CI, 0.91 to 1.01; P =.09). The adjusted difference in ICU LOS was similar for admissions from July through September in major (0.3%; 95% CI, -0.7% to 1.2%; P =.61) and minor (0.2%; 95% CI, -0.9% to 1.4%; P =.69) teaching hospitals, but was somewhat shorter in nonteaching hospitals (-0.8%; 95% CI, -1.4% to -0.1%; P =.03). Results were similar when individual months and academic years were examined separately, and in stratified analyses of surgical and nonsurgical patients. CONCLUSIONS: We found no evidence to support the existence of a July phenomenon in ICU patients. Future studies should examine organizational factors that allow hospitals and residency programs to compensate for inexperience of new housestaff early in the academic year. PMID- 12911647 TI - Medical errors related to discontinuity of care from an inpatient to an outpatient setting. AB - OBJECTIVE: To determine the prevalence of medical errors related to the discontinuity of care from an inpatient to an outpatient setting, and to determine if there is an association between these medical errors and adverse outcomes. PATIENTS: Eighty-six patients who had been hospitalized on the medicine service at a large academic medical center and who were subsequently seen by their primary care physicians at the affiliated outpatient practice within 2 months after discharge. DESIGN: Each patient's inpatient and outpatient medical record was reviewed for the presence of 3 types of errors related to the discontinuity of care from the inpatient to the outpatient setting: medication continuity errors, test follow-up errors, and work-up errors. MEASUREMENTS: Rehospitalizations within 3 months after the initial postdischarge outpatient primary care visit. MAIN RESULTS: Forty-nine percent of patients experienced at least 1 medical error. Patients with a work-up error were 6.2 times (95%confidence interval [95% CI], 1.3 to 30.3) more likely to be rehospitalized within 3 months after the first outpatient visit. We did not find a statistically significant association between medication continuity errors (odds ratio [OR], 2.5; 95%CI, 0.7 to 8.8) and test follow-up errors (OR, 2.4; 95%CI, 0.3 to 17.1) with rehospitalizations. CONCLUSION: We conclude that the prevalence of medical errors related to the discontinuity of care from the inpatient to the outpatient setting is high and may be associated with an increased risk of rehospitalization. PMID- 12911648 TI - Improving the physical diagnosis skills of third-year medical students: a controlled trial of a literature-based curriculum. AB - OBJECTIVE: To determine if a literature-based physical diagnosis curriculum could improve student knowledge, skill, and self-confidence in physical diagnosis. DESIGN: Prospective controlled trial of an educational intervention. SETTING: Required internal medicine clerkship for third-year medical students at Brown Medical School. PARTICIPANTS: Third-year medical students who completed the internal medicine clerkship during the academic year 1999-2000: 32 students at 1 clerkship site received the intervention; a total of 50 students at 3 other clerkship sites served as controls. INTERVENTION: Physical diagnosis curriculum based on 8 articles from the Journal of the American Medical Association's Rational Clinical Examination series. Intervention students met weekly for 1 hour with a preceptor to review each article, discuss the sensitivity and specificity of the maneuvers and findings, and practice the techniques with an inpatient who agreed to be visited and examined. MEASUREMENTS AND MAIN RESULTS: Physical diagnosis knowledge for the 8 topics was evaluated using a 22-item multiple choice question quiz, skill was evaluated using trained evaluators, and self confidence was assessed using an end-of-clerkship survey. Intervention students scored significantly higher than the control group on the knowledge quiz (mean correct score 70% vs 63%, P =.002), skills assessment (mean correct score 90% vs 54%, P <.001), and self-confidence score (mean total score 40 vs 35, P =.003), and they expressed greater satisfaction with the physical diagnosis teaching they received in the clerkship. CONCLUSION: This physical diagnosis curriculum was successful in improving students' knowledge, skill, and self-confidence in physical diagnosis. PMID- 12911649 TI - Morbidity and mortality conference: a survey of academic internal medicine departments. AB - This study sought to determine the prevalence and characteristics of morbidity and mortality conferences (M&MCs) in U.S. internal medicine training programs. Two hundred ninety-five of 416 (71%) surveys were returned. Ninety percent of programs have an M&MC. Most meet monthly, have a designated leader, and entail case discussions of 3 or fewer patients. Cases are selected on the basis of unexpected bad outcomes, teaching value, and to a lesser extent, suspected medical error. Two thirds of the sites use M&MCs to meet administrative requirements for quality assurance. M&MC, while prevalent in internal medicine training programs, has a heterogeneity of focus. Hence, the goals and role of the conference, as judged by this survey, do not appear to be well defined and may warrant further clarification. PMID- 12911650 TI - Patient perspectives of medical confidentiality: a review of the literature. AB - OBJECTIVE: To lay the groundwork for a better understanding of patient views on medical confidentiality. DESIGN: Studies were found by searching medline, bioethicsline, and selected bibliographies. Articles concerning physician perspectives or implications of legal and administrative regulations were excluded. Only peer-reviewed journal articles reporting original research on patients' confidentiality views and conduct were included. MAIN RESULTS: Many patients are unaware of or misunderstand their legal or ethical right to medical confidentiality protections, which leads them to both over- and underestimate confidentiality protections. The possibility that medical information might be revealed, intentionally or not, to acquaintances in a clinic or other social community troubles patients as much as information release to insurers or employers. A significant minority of patients distrust confidentiality protections, leading some to report that they delay or forgo medical care. If doubtful that confidentiality will be upheld, patients will act independently to protect information. CONCLUSIONS: Our review found a wider variety of understandings and beliefs about medical confidentiality among patients than are often indicated in the writings of practitioners or legal experts. As medical confidentiality regulations evolve, these differences need to be recognized and accounted for in interactions between practitioners and patients. PMID- 12911651 TI - What is clinical empathy? AB - Patients seek empathy from their physicians. Medical educators increasingly recognize this need. Yet in seeking to make empathy a reliable professional skill, doctors change the meaning of the term. Outside the field of medicine, empathy is a mode of understanding that specifically involves emotional resonance. In contrast, leading physician educators define empathy as a form of detached cognition. In contrast, this article argues that physicians' emotional attunement greatly serves the cognitive goal of understanding patients' emotions. This has important implications for teaching empathy. PMID- 12911652 TI - Seeing error through new lenses. PMID- 12911653 TI - Research on patient-clinician relationships: celebrating success and identifying the next scope of work. PMID- 12911654 TI - Irritable bowel syndrome: a global view. PMID- 12911655 TI - Of blood and guts: association between Helicobacter pylori and the gastric microcirculation. AB - The relative importance of acid in Helicobacter pylori (H. pylori) ulcer pathogenesis is in doubt, with possibilities existing that other contributing factors may be involved. Vascular insufficiency may lead to the development of ischemic lesions or ulcers within the gastric mucosa. H. pylori produces a striking inflammatory response following infection and one of the major components of gastrointestinal inflammation is alterations in the vascular structure and function. This suggests that the microcirculation may be a key target of H. pylori-released factors. Recent evidence has accumulated to suggest that H. pylori can affect a number of microcirculatory variables including blood flow, leukocyte activity and also induce changes in the endothelial lining of the vessels themselves. The majority of these findings have been described by employing the technique of fluorescent in vivo microscopy, which allows direct, dynamic and real time observations of the microcirculation to be made. A universal feature of these experimental studies has been the formation of circulating or adherent platelet aggregates. It is now recognized that platelets participate in the inflammatory response by acting as a potent source of inflammatory mediators and modulating the activity of other inflammatory cells. Circulation of platelet emboli may be of cause for concern, especially as a number of studies have demonstrated an association between H. pylori infection and coronary heart disease. The present review highlights the major findings from these studies and proposes an important role for the gastric microcirculation in the pathophysiology of H. pylori-induced injury. PMID- 12911656 TI - Genotypes of Helicobacter pylori in children with upper abdominal pain. AB - BACKGROUND: Studies related to genotypes of Helicobacter pylori infection and upper abdominal pain (UAP) in children are scarce all over the world. We prospectively analyzed the association between H. pylori infection and UAP in our study group of children and evaluated the vacA genotypes associated with this disorder. METHODS: We investigated 34 children with UAP (group I) and another 110 children as controls without UAP (group II) for H. pylori infection, using antral biopsies by culture, rapid urease tests, histopathology and ureA polymerase chain reaction (PCR). Genotyping was performed using specific cagA and vacA primers on 52 H. pylori strains (group I = 21; 18 boys, mean age 10.33 +/- 2.47 years; group II = 31; boys 21, mean age 9.63 +/- 2.51 years). RESULTS: A significant association between H. pylori infection and UAP was observed when compared to controls (21/34; 61.8%vs 31/110; 28.2%; P = 0.0004). cagA positive H. pylori strains were detected in 20 (95.2%) children with UAP and in 28 (90.3%) controls. Mixed infection was detected in 25% of children with no significant difference between the groups. On univariate regression analysis, s1a, m1 alleles and s1a/m1 genotype of vacA had significant associations with UAP (P = 0.018, 0.015 and 0.007, respectively), while s2 and m2 alleles and the s2/m2 genotype were significantly more frequent in controls (P = 0.034, 0.001 and 0.034, respectively). CONCLUSIONS: H. pylori infection is strongly associated with UAP in children and a significantly higher proportion have s1a, m1 alleles and s1a/m1 genotype. The negative associations of vacA s2, m2 alleles and s2/m2 genotype with UAP indicate that they are unlikely to have an important role in this disorder. PMID- 12911657 TI - Effect of Helicobacter pylori infection and low-dose aspirin use on iron stores in the elderly. AB - BACKGROUND AND AIMS: Helicobacter pylori infection has been associated with lower ferritin levels in some adult populations, but subsets of potentially higher risk subjects, such as the elderly have not been examined. The aim of the present study was to determine the impact of H. pylori infection and low-dose aspirin use on iron stores in a well elderly population. METHODS: Consecutive subjects aged 65 years or older attending day care facilities were studied. Each subject was assisted in completing a medical questionnaire. Serum ferritin, hemoglobin, mean corpuscular volumes (MCV) and IgG antibodies (ELISA) against H. pylori were measured. RESULTS: In 220 subjects (age 75 +/- 8 years), 42% were H. pylori seropositive (male 41%, female 44%). The median (IQR) ferritin level (ug/L) was higher in men 149 (89-280) than women 94 (54-161), p < 0.002. The ferritin levels were not different in H. pylori positive males 151 (105-283) compared with H. pylori negative males 145 (72-249), or H. pylori positive females 93 (60-142) compared with H. pylori negative females 97 (45-149). This relationship was not altered when controlled for non-steroidal anti-inflammatory agents (NSAID) use (9% of subjects), alcohol or dietary iron intake. Low-dose aspirin use was common (28%), but did not have an independent impact on iron stores: male users 181 (95 248), non-users 145 (86-284); female users 92 (43-162), non-users 95 (62-163). However, in female aspirin users, H. pylori infection was associated with significantly lower ferritin levels: 65 (43-112) compared with uninfected subjects 103 (41-180), p < 0.04. CONCLUSIONS: In this well elderly population, the combination of H. pylori infection and low-dose aspirin use was associated with significantly lower serum ferritin concentrations in females. This difference supports the observation that H. pylori, even in asymptomatic subjects, may be a stressor of iron stores. PMID- 12911658 TI - Combination therapy of ecabet sodium and cimetidine compared with cimetidine alone for gastric ulcer: prospective randomized multicenter study. AB - BACKGROUND AND AIM: Little is known about the clinical efficacy of co-therapy of ecabet sodium, a mucoprotective agent, and a histamine H2-receptor antagonist. The aim of the present study was to assess its additive benefit in combination with cimetidine for gastric ulcer. METHODS: In this prospective randomized study, after gastric ulcer was confirmed by endoscopy, 200 patients in 47 hospitals received either ecabet sodium 1 g b.i.d and cimetidine 400 mg b.i.d. (EC), or cimetidine 400 mg b.i.d. alone (C) for 8 weeks. Healing was examined by endoscopy at 4 and 8 weeks. RESULTS: Of the intention-to-treat (ITT) population (EC, 103; C, 97), 181 patients comprised the per protocol (PP) analysis (EC, 93; C, 88). At 4 weeks, healing rates were significantly higher in the EC group (60%) than in the C group (36%) ( p < 0.01). At 8 weeks, those by the ITT and PP analyses were 82% (EC) versus 58% (C), and 90% (EC) versus 64% (C), respectively ( p < 0.01 and p < 0.001). Symptom relief rates (EC vs C) at 2, 4 and 8 weeks were 73%versus 47% ( p < 0.01), 89%versus 66% ( p < 0.001), and 97%versus 73% ( p < 0.001), respectively. Significant additive effects of ecabet sodium were observed in patients aged 60 years or older, with solitary and medium to large ulcer, and without smoking or drinking habits. No adverse effects were critical. CONCLUSION: Ecabet sodium significantly augmented gastric ulcer healing and symptom relief by cimetidine, especially in the elderly. PMID- 12911659 TI - Comparison of the effect on intragastric pH of a single dose of omeprazole or rabeprazole: which is suitable for on-demand therapy? AB - BACKGROUND AND AIMS: An ideal medication for heartburn should have the rapid onset of action needed for on-demand treatment. However, assessment of the onset of action of proton pump inhibitors has been largely subjective. We compared the inhibitory effect on gastric acid secretion of a single oral dose of omeprazole with that of rabeprazole. METHODS: Fourteen Helicobacter pylori-negative men participated in this randomized, double-masked, two-way cross-over study. Intragastric pH was monitored continuously for 6 h after a single, randomly assigned 20 mg oral dose of either omeprazole or rabeprazole. After a 7-day washout period, the other drug was administered. Each patient's S-mephenytoin 4' hydroxylase (CYP2C19) genotype was determined by polymerase chain reaction restriction fragment length polymorphism. RESULTS: Intragastric pH and pH holding time did not differ between treatments when the data were analyzed for the whole group without stratifying for CYP2C19 status. In CYP2C19 homozygous and heterozygous extensive metabolizers (10 subjects), rabeprazole maintained the intragastric at pH > 3 and> 4 for longer than omeprazole during both the 5 and 6 h study periods, and the average pH during the 6 h study period was higher with rabeprazole than with omeprazole. In these extensive metabolizers, rabeprazole maintained the pH > 2,> 3,> 3.5 and> 4 for longer during the 6 h study period than did omeprazole. CONCLUSIONS: In H. pylori-negative men who are CYP2C19 homozygous or heterozygous extensive metabolizers, the intragastric pH after a single dose of 20 mg rabeprazole is higher during first 5-6 h than that after a single dose of 20 mg omeprazole. PMID- 12911660 TI - Inducible nitric oxide synthase activity induced by sodium chloride solution prolongs luminal pH elevation in rat and mouse stomachs. AB - BACKGROUND: Sodium chloride (NaCl) is a strong promoter of gastric cancer. We hypothesized that inducible nitric oxide synthase (iNOS) induced by NaCl may be involved in its promoting effects. We investigated iNOS expression by hypertonic NaCl solutions and its pathophysiological roles in the gastric mucosa of rats and mice. METHODS: iNOS mRNA and protein expressions were examined in the rat and mouse gastric mucosa after intragastric administration of NaCl solution by northern blot, reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry. The effect on luminal pH by iNOS activity was assessed using aminoguanidine, a potent iNOS inhibitor, and iNOS gene deficient (iNOS-/-) mice. RESULTS: iNOS expression was detected at concentrations higher than 1.7 M, mainly in the cells infiltrating the damaged mucosa of rats. Administration of a higher than 3.4 M NaCl solution elevated luminal pH of the rat stomach remarkably, enabling bacterial overgrowth and dramatically increasing iNOS expression (n = 5 per concentration). Pretreatment with ampicillin (p.o), an antibiotic, attenuated the iNOS expression in duplicate experiments. Co-treatment with aminoguanidine (s.q) accelerated recovery of elevated luminal pH at 8 h and 16 h or 24 h after administration of 3.4 M (n = 8) and 5.2 M NaCl solution (n = 5), respectively. iNOS expression and luminal pH elevation were also observed in mice stomachs after administration of 3.4 M NaCl solution. The elevated luminal pH of iNOS-/- mice stomachs after the administration of NaCl solution was significantly lower at 6 h (n = 7) and at 9 h (n = 11), compared to that of wild type mice (n = 9 and 10, respectively). CONCLUSIONS: Hypertonic NaCl solutions induced iNOS expression in the gastric mucosa. iNOS activity prolonged the elevation of the luminal pH, potentially leading to bacterial overgrowth, which in turn enhanced iNOS expression. This vicious cycle might be related to the promoting effect of NaCl. PMID- 12911661 TI - p53 expression, K-ras gene mutation and microsatellite instability in gastric B cell lymphomas. AB - BACKGROUND AND AIMS: Genetic mechanisms involved in the development of gastric B cell lymphomas remain unclear. The aim of the present study was to clarify the roles of mutations of the p53 and K-ras genes, and microsatellite instability (MSI) in the development of gastric B-cell lymphomas. METHODS: We investigated p53 immunoreactivity, mutations of the K-ras gene, and MSI in 27 gastric marginal zone B-cell lymphomas of mucosa-associated lymphoid tissue type (MZBCL) and 24 diffuse large B-cell lymphomas (DLBCL). p53 immunoreactivity was examined using a monoclonal antibody, DO-7. Mutation of the K-ras gene was detected by polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) analysis. MSI was examined at five microsatellite loci with a microsatellite assay. Cases were classified as having high-frequency MSI (MSI-H) (>/= 2 loci showing instability), low-frequency MSI (MSI-L) (only one locus showing instability), or as microsatellite stable. RESULTS: p53 immunoreactivity was detected in 1 of 16 (6%) MZBCL and 8 of 19 (42%) DLBCL. Frequency of p53 immunoreactivity in DLBCL was significantly higher than that in MZBCL (P = 0.018). MSI-H was detected only in 1 of 20 (5%) DLBCL. None of the cases examined showed mutation of the K-ras gene. CONCLUSIONS: These data suggest that mutations of the p53 gene may play an important role in the development of gastric DLBCL, and that mutations of the K ras gene and MSI may be involved in little part of the development of gastric B cell lymphomas. PMID- 12911662 TI - Predicting mortality in patients with cirrhosis of liver with application of neural network technology. AB - BACKGROUND: Prediction of mortality from cirrhosis is important in planning optimal timing of liver transplantation and other interventions. We evaluated the role of the Artificial Neural Network (ANN), which uses non-linear statistics for pattern recognition in predicting one-year liver disease-related mortality using information available during initial clinical evaluation. METHODS: The ANN was constructed using software with data from a training set (n = 46) selected at random from a cohort of adult cirrhotics (n = 92). After training, validation was performed in the remaining patients (n = 46) whose outcome in terms of one-year mortality was unknown to the network. The performance of ANN was compared to those of a logistic regression model (LRM) and Child-Pugh's score (CPS). Death (related to cirrhosis/its complications) within one year of inclusion was the outcome variable. The ANN was also tested in an external validation sample (EVS, n = 62) from another hospital. RESULTS: Patients in the EVS were younger (mean age, 41 vs 45 years), infrequently of alcoholic etiology (5% vs 49%), had less severe disease (mean CPS 6.6 vs 10.8), and had lower one-year mortality (13 vs 46%). In the internal validation sample, ANN's accuracy was 91%, sensitivity 90% and specificity 92% in prediction of one-year mortality; area under the receiver operating characteristic (ROC) curve was 0.94. The performance of the LRM (accuracy 74%) and the CPS (accuracy 55%) was significantly worse than ANN (P < 0.05, McNemar's test). Despite differences in the characteristics of the two groups, the ANN performed fairly well in the EVS (accuracy of 90%, area under curve 0.85). CONCLUSIONS: ANN can accurately predict one-year mortality in cirrhosis and is superior to CPS and LRM. PMID- 12911663 TI - Analysis of major histocompatibility complex and CTLA-4 alleles in Brazilian patients with primary biliary cirrhosis. AB - BACKGROUND AND AIMS: Predisposition to primary biliary cirrhosis (PBC) has been classically linked to HLA-DRB1 locus. However, the presence of the HLA-DRB1*08 antigen has been reported in less than one-third of PBC patients from Northern Europe and Japan. Recently, polymorphisms in the tumor necrosis factor alpha (TNFA) gene promoter at position -308 and in exon 1 of the cytotoxic T lymphocyte antigen-4 (CTLA-4) gene at position 49 have been associated with susceptibility to PBC in Caucasians. In addition, the presence of HLA-DRB1*08 and the TNFA*1 allele was also linked to progression to end-stage liver disease. The aims of the present study were to investigate the frequencies of HLA-DR and DQ antigens and TNFA and CTLA-4 alleles in PBC patients from a different genetic background, as well as to assess the role of TNFA alleles and HLA-DR antigens in disease progression. METHODS: Determination of HLA-DRB1, DQB1, TNFA and CTLA-4 alleles was performed in patients with PBC and healthy controls using polymerase chain reaction-based techniques. RESULTS: Frequencies of HLA-DR and DQ antigens were similar in PBC patients and healthy controls. Accordingly, no association between TNFA and CTLA-4 alleles was observed in PBC patients. The histological stage at admission of patients with PBC also showed no correlation with HLA antigens and TNFA and CTLA-4 alleles. CONCLUSIONS: Susceptibility to PBC in Brazil is not associated with HLA-DR and DQ antigens and CTLA-4 genotypes. TNFA alleles were not shown to influence disease progression. PMID- 12911664 TI - Concurrent pancreatic ductal changes in alcoholic liver disease. AB - BACKGROUND: Alcohol is a common etiological factor in both liver disease and chronic pancreatitis, but in a single individual it does not usually produce clinically significant disease in both organs. We assessed the prevalence of pancreatic ductal changes in patients presenting with alcoholic liver disease of different stages. METHODS: Forty-six patients with alcoholic liver disease were included in the present study. Liver biopsy was performed in patients with normal coagulogram. Endoscopic retrograde pancreatogram was performed in all patients and changes in chronic pancreatitis were noted. RESULTS: Of the 46 patients with alcoholic liver disease, 31 had cirrhosis of the liver, nine had fatty liver and two patients had alcoholic hepatitis. Twenty (43.47%) patients had features of chronic pancreatitis on endoscopic retrograde pancreatogram and these consisted of minimal pancreatitis changes in 10 patients, moderate changes in nine patients and advanced changes in one patient. There was no difference in the prevalence of pancreatitis changes in cirrhotics in comparison to non-cirrhotics. There was no correlation between the amount and length of alcohol intake and changes in pancreatitis. CONCLUSION: Pancreatic ductal changes on endoscopic retrograde pancreatogram are common in patients with alcoholic liver disease. PMID- 12911665 TI - Experimental studies on morphological changes of microcirculation of DMN-induced liver cirrhosis after normothermic ischemia with charge-coupled device microscope. AB - AIM: The purpose of the present experiment was to find indices for intraoperative ischemic-reperfusion injury in the cirrhotic liver. METHODS: One percent dimethylnitrosamine (DMN) was administered intraperitoneally to liver cirrhosis (LC) groups of Wister rats on three consecutive days of each week for a period of four weeks. The rats were divided into a N60 group with 60 minute ischemia in normal livers, a LC60 group with 60 minute ischemia in cirrhotic livers, and a LC30 group with 30 minute ischemia in cirrhotic livers. Digital videotapes recorded with a pencil lens-probe charge-coupled device (CCD) microscope were analyzed with NIH Image software. In zone 3, the sinusoid diameter (SD) was measured and the volume fraction (Vv) of zone 3 was calculated in preischemia and after 10, 20, 30, and 60 min of reperfusion. At the same time, bile flow was measured. RESULTS: The SD was significantly shorter in the cirrhotic liver groups than in the normal liver group at each point. The Vv after 60 min of reperfusion was significantly smaller in the LC60 group, with a survival rate of 0%, than in the LC30 group which had a survival rate of 67%. However, there was no significant difference in bile flow after 60 min of reperfusion in the LC30 and LC60 groups. Therefore, the Vv is suggested to be the better index for viability after ischemic-reperfusion. CONCLUSION: SD and Vv indicate microcirculatory differences and indices in the normal and cirrhotic livers in preischemia during reperfusion. PMID- 12911666 TI - Computed tomography angiographic findings in hepatocellular carcinoma less than 2 cm detected during follow-up in 29 patients. AB - AIMS: The early stage of hepatocarcinogenesis is not well understood pathologically and clinically. The present study was designed to define small (early) hepatocellular carcinoma (HCC) angiographically using the angio-helical computed tomography (CT) system. METHODS: Arterial portography CT and hepatic arteriography CT were carried out in 29 patients in whom small HCC < or = 20 mm was detected during follow-up. RESULTS: There were 17 males and 12 females, aged 47 to 85 years. The offending virus was hepatitis B in four, hepatitis C in 24 and no virus marker in one case. The follow-up period varied from less than a year to 17 years, averaging 6.4 years. The underlying disease was cirrhosis in 12 and chronic hepatitis in 17 cases. The mass was solitary in 16 and multiple in 13 cases, while the size of the mass ranged from 8-20 mm. All lesions were low in attenuation on arterial portography CT, and in 23 of 30 lesions hepatic arteriography CT showed high attenuation, suggesting arterial blood supply. In the remaining 7 cases, lesions were perhaps in the transition from portal to arterial. CONCLUSIONS: It was concluded that HCC develops frequently in a liver with chronic hepatitis, often muticentrically, and that early HCC lesions are more often overt HCC already with arterial blood supply, rather than extremely well-differentiated supplied by the portal vein as generally believed. PMID- 12911667 TI - Differential expression of CCR5 and CRTH2 on infiltrated cells in colonic mucosa of patients with ulcerative colitis. AB - BACKGROUND AND AIM: The pathogenesis of ulcerative colitis (UC) is unclear, but abnormal infiltration of T lymphocytes in the colonic mucosa has been implicated in the mucosal tissue damage. The abnormal cytokine production because of a T helper (h)1/Th2 imbalance may play an important role in continuing inflammation in the colonic mucosa. In the present study, the expression of chemokine receptor 5 (CCR5) as a Th1 marker and a chemoattractant receptor-homologs molecule expressed on Th2 cells (CRTH2) were investigated in order to analyze impaired Th1/Th2 responses in the colonic mucosa of UC patients. METHODS: Tissue samples were obtained by colonic biopsies from patients with UC or colonic polyps, with informed consent. Immunohistochemical analysis was performed on periodate, lysine paraformaldehyde-fixed serial cryostat sections using the labeled streptavidin biotin method. Monoclonal antibodies against CD4, CCR5 or CRTH2 were used as primary antibodies. The number of cells expressing CD4, CCR5 or CRTH2 per unit area was calculated by using an image analyzer. RESULTS: In the patients with UC, the numbers of CD4- and CCR5-positive cells were significantly increased in inflamed mucosa, and appeared to be correlated with the disease activity. The infiltration of CRTH2-positive cells was predominantly observed in the mildly inflamed or the margin of inflamed mucosa of UC patients. CONCLUSION: There is a possibility that Th1 responses significantly occur in colonic mucosa with severe inflammation, while Th2 responses mainly occur with mild inflammation in UC patients. The Th1/Th2 imbalance in colonic mucosa may be related to the disease progression of UC. PMID- 12911668 TI - Gamma-amino-butyric acid immunoreactivity in intramucosal colonic tumors. AB - BACKGROUND AND AIM: The level of gamma-amino-butyric acid (GABA) is reported to be increased in colon cancer. Moreover, data suggests that GABA plays a role in the proliferation or maturation of some types of cells. We examined the expression of GABA in intramucosal colonic tumors to clarify the relation between GABA and the degree of atypia. METHODS: Paraffin sections were prepared from 56 protruded-type colonic neoplasms, which were classified as intramucosal adenocarcinoma (AC), adenoma with severe atypia (ASA), or adenoma with mild to moderate atypia (AMA). Expression of GABA was investigated immunohistochemically, and GABA immunoreactivity was compared to the staining patterns of carcinoembryonic antigen (CEA) and cancer-associated antigen (CA19-9) which were classified into three categories. RESULTS: Intense GABA immunoreactivity was observed in 73.7%, 54.6%, 13.3%, and 5.4% of AC, ASA, AMA, and normal mucosa specimens, respectively. Kendall's correlation coefficient between GABA immunoreactivity and the degree of atypia was 0.447. Strong, positive CEA staining (pattern 3) was observed in 57.9%, 36.3%, and 13.3% of AC, ASA, and AMA specimens, respectively. Strong, positive CA19-9 staining was observed: 26.3%, 9.1% and 0%, respectively. In AC and ASA, the proportion of glands with strong GABA immunoreactivity was greater than the proportion of glands that were strongly positive for CA19-9. CONCLUSION: GABA may be useful as a tumor marker in combination with other tumor markers such as CEA and CA19-9. PMID- 12911669 TI - Irinotecan causes severe small intestinal damage, as well as colonic damage, in the rat with implanted breast cancer. AB - BACKGROUND AND AIMS: Irinotecan (CPT-11) is a chemotherapeutic drug for cancer that causes severe diarrhea by an uncertain mechanism. The aim of the present study was to investigate the time-course of apoptosis and whole intestinal damage after irinotecan to further elucidate the mechanism behind the diarrhea. METHODS: Groups of breast cancer-bearing dark agouti (DA) rats were treated with 100, 150 or 200 mg/kg doses of irinotecan or vehicle control daily for two days, and killed at 6, 24, 72 or 96 h after treatment. Apoptosis and morphometry were examined in both the small and large intestines. Histopathology and goblet cell numbers were recorded. Data were analyzed using the Peritz' F-test. RESULTS: Irinotecan increased apoptosis and caused villous atrophy and crypt hypoplasia in the small intestine, and increased apoptosis, crypt hypoplasia, crypt dilation and mucus secretion in the large intestine. Irinotecan at 100 and 150 mg/kg caused crypt hypoplasia at 6 and 24 h, with rebound hyperplasia at 72 and 96 h. At 200 mg/kg, irinotecan caused a more pronounced crypt hypoplasia earlier and all animals died by 96 h. Apoptosis peaked at 6 h and remained elevated over the remainder of the time-points. This was not dose-dependent. Irinotecan at all doses altered colonic, but not jejunal, goblet cells. Irinotecan increased colonic mucus secretion. CONCLUSIONS: We conclude that irinotecan causes diarrhea by inducing apoptosis and hypoproliferation in both the small and large intestines, and causes colonic damage with changes in goblet cells and mucin secretion. PMID- 12911670 TI - Hepatobiliary and pancreatic: Woman with acute intermittent porphyria. PMID- 12911671 TI - Gastrointestinal: Antral diaphragm. PMID- 12911673 TI - Esophageal xanthoma: report of two cases and a review of the literature. AB - So far, three cases of esophageal xanthoma have been reported. We describe here endoscopic, microscopic and immunohistochemical findings of two new cases of this rare condition, and a review of five cases of esophageal xanthoma, including our cases. Esophageal xanthomas endoscopically show yellow granular spots or a slightly elevated lesion, and are similar to an ectopic sebaceous gland. Microscopically, an aggregate of foamy histiocytes (lipid islands) is seen immediately beneath the squamous epithelium, particularly between the rete ridges. Immunohistochemically, xanthoma cells are positive for CD68, which indicate a histiocytic origin. The etiology of esophageal xanthoma remains unclear. We emphasize that esophageal xanthomas may have been erroneously diagnosed as an ectopic sebaceous gland on endoscopic examination. PMID- 12911674 TI - Successful living related liver transplantation for adult Still's disease. PMID- 12911675 TI - Acute pancreatitis due to hydrocortisone in a patient with ulcerative colitis. PMID- 12911676 TI - Does paracetamol cause asthma? PMID- 12911677 TI - Prescription drug samples--does this marketing strategy counteract policies for quality use of medicines? AB - Prescription drug samples, as used by the pharmaceutical industry to market their products, are of current interest because of their influence on prescribing, and their potential impact on consumer safety. Very little research has been conducted into the use and misuse of prescription drug samples, and the influence of samples on health policies designed to improve the rational use of medicines. This is a topical issue in the prescription drug debate, with increasing costs and increasing concerns about optimizing use of medicines. This manuscript critically evaluates the research that has been conducted to date about prescription drug samples, discusses the issues raised in the context of traditional marketing theory, and suggests possible alternatives for the future. PMID- 12911678 TI - Comparative study on two kidney graft rinsing and preservation solutions in terms of the post-transplantation risk of delayed graft function and cost. AB - OBJECTIVE: To determine whether Belzer solution (Viaspan, Bristol-Myers Squibb, Brussels, Belgium), which is more expensive than Eurocollins solution, was better at preventing delayed graft function (DGF) and whether it was cost-effective as it could potentially reduce post-transplantation complications. METHOD: The risk of occurrence of complications associated with the use of these two rinsing and preserving solutions was estimated from a survey of 106 patients undergoing renal transplantation between 1 January 1993 and 31 March 1998. Both efficacy and adverse outcomes were recorded along with the costs directly associated with the transplantation procedure in the hospital setting: hospitalization, rinsing and preserving solutions, medical and technical interventions and diagnostic tests. RESULTS: For the 45 kidney grafts rinsed and preserved with Eurocollins (strategy S1: n1 = 45) the cost/graft was estimated at 40 euros. With Viaspan (strategy S2: n2 = 61) the corresponding cost/graft was 424 euros. Logistic regression analysis showed that Viaspan was better than Eurocollins solution (ebeta = 0.437; P = 0.05) in preventing DGF. Overall, S2 was less expensive than S1, from the hospital's perspective. The mean difference per patient was 278 euros, which amounts to a saving of 2% of the total cost per renal transplantation. For rinsing and preserving kidney grafts Belzer solution is therefore preferable to Eurocollins solution. PMID- 12911679 TI - Genetic polymorphism of CYP2D6 in Chinese subjects in Malaysia. AB - BACKGROUND: Although Malaysian Chinese share an origin with the mainland Chinese, their evolution has been influenced by intermarriages. With a gene such as CYP2D6, which is highly polymorphic, it is expected that the Malaysian Chinese would exhibit a polymorphism profile different from those of the Chinese populations in other geographical locations. OBJECTIVE: To study the genotype distribution of CYP2D6 among the Chinese people in Malaysia. METHOD: We obtained DNA from 236 Chinese individuals in Malaysia and used PCR-based methods to identify any common CYP2D6 alleles. RESULTS: A total of 236 subjects were enrolled and were successfully genotyped. Malaysian Chinese were relatively heterogeneous in terms of their CYP2D6 genotypes with nine genotypes recorded. CYP2D6*4, *5, *9, *10 and *17 were detected with the most common genotype being *1/*10. No subject had genotypes that predicted poor metabolic activity. However, 40% showed genotypes (e.g. CYP2D6*10/*10, *17, *4 and *9 and *9/*9) that predicted an intermediate metabolizer phenotype. Another subject carried the defective CYP2D6*17 allele and six carried the defective CYP2D6*9 allele. Both these alleles have not been reported in other earlier Chinese studies. CONCLUSION: This study revealed that, in terms of CYP2D6 polymorphism, Malaysian Chinese were a heterogeneous group of people. Although sharing some similarities with other Orientals, they also seemed to have some notable differences. The alleles CYP2D6*4, *5, *9, *10 and *17 were all detected. CYP2D6*3 was however absent. PMID- 12911680 TI - The safety of spironolactone treatment in patients with heart failure. AB - INTRODUCTION: Spironolactone is increasingly being used in the treatment of heart failure. However, it has been associated with cases of hyperkalaemia. The common use of angiotensin-converting enzyme (ACE) inhibitors and angiotensin-2 receptor (AT2) antagonists in heart failure increases the risk of hyperkalaemia. OBJECTIVE: To identify the risk of spironolactone withdrawal, hyperkalaemia and severe hyperkalaemia in patients prescribed spironolactone together with either an ACE inhibitor or an AT2 antagonist. METHODS: Retrospective identification and analysis of patients co-prescribed an ACE inhibitor or an AT2 antagonist with spironolactone. Patients' records were linked to their biochemical results and the doses of spironolactone, ACE inhibitor and AT2 antagonists received by them. RESULTS: We found that a higher proportion of patients in our cohort stopped taking spironolactone compared with the Randomised Aldactone Evaluation Study and a higher proportion developed hyperkalaemia, a predicted adverse effect of spironolactone combination with an ACE inhibitor or an AT2 antagonist. Patients with diabetes mellitus and those with a haematocrit below 0.36, were more likely to develop hyperkalaemia than those without these traits. CONCLUSIONS: Spironolactone is a common cause of hyperkalaemia when used in combination with either an ACE inhibitor or an AT2 antagonist. This reinforces the need for care when extrapolating the results of clinical trials to daily clinical practice. PMID- 12911681 TI - The impact of therapeutic doses of paracetamol on serum total antioxidant capacity. AB - INTRODUCTION: A link between regular paracetamol intake and asthma in adults has recently been postulated. Detoxification of paracetamol may deplete stores of glutathione, which is one of the major antioxidants present in the lung. A reduced source of glutathione in the lung may lead to increased oxidative damage to the epithelium and hence increased frequency and severity of asthma attacks in susceptible individuals. AIM OF STUDY: This study aimed to determine whether regular intake of maximum therapeutic doses of paracetamol reduced serum antioxidant capacity in healthy volunteers. METHODS: Fifteen young healthy volunteers (nine men, six women, mean age 21.3 years, range 19-32) took maximum therapeutic doses of paracetamol (1 g four times a day) for 14 days. On days 0 and 14 blood samples were taken at baseline and hourly for a period of 4 h following ingestion of 1 g paracetamol. Single venous blood samples were collected 1 h after ingestion of 1 g paracetamol on days 4, 7 and 10. Blood samples were analysed for serum paracetamol concentration and total antioxidant capacity. RESULTS: Mean total antioxidant capacity was significantly reduced over the 3-h post-dosing on both days 0 and 14 (P < 0.01). The results from days 4, 7 and 10 showed a trend towards reduced antioxidant activity over time. On day 14 values were consistently lower compared with the corresponding times on day 0 (P < 0.01 at 0, 1, 3 and 4 h, P < 0.05 at 2 h). CONCLUSIONS: Chronic ingestion of maximum therapeutic doses of paracetamol depletes serum antioxidant capacity in healthy volunteers in as few as 14 days, possibly by a reduction in glutathione. This may have implications for analgesic use in asthmatic individuals. Further studies are now required to assess the impact of paracetamol on antioxidant defences in the lung. PMID- 12911682 TI - Preventing drug-related morbidity--the development of quality indicators. AB - OBJECTIVE: To describe the process that has been undertaken to (i) validate further UK indicators for preventable drug-related morbidity (PDRM) generated by a previous study and (ii) develop additional new indicators derived from UK clinical practice. DESIGN: A two-round Delphi questionnaire survey. SETTING: A UK study based in primary care. PARTICIPANTS: A purposively selected sample of primary care pharmacists (n = 9) and general practitioners (n = 8). OUTCOME MEASURES: The establishment of consensus among the panel as to whether an indicator represented a PDRM. RESULTS: The pre-defined level of consensus was achieved for 24 indicators (59%; 24 of 41). CONCLUSIONS: We have identified that although mechanisms exist for identifying 'the outcome' component of PDRM indicators, identifying the 'pattern of care' that is needed to prevent their occurrence is far more challenging. Nonetheless, we have taken considerable steps along the path of validating such indicators. Future operationalization in a general practice setting should help to facilitate improvements in medicine management in primary care and ultimately benefit patients. PMID- 12911683 TI - Paracetamol poisoning--impact of pack size restrictions. AB - BACKGROUND: Pack sizes of paracetamol available to the public were reduced by legislation in 1998. OBJECTIVE: To study the effect of the legislation on paracetamol poisoning related admissions to an accident and emergency department and to a tertiary referral liver unit. METHOD: Data were obtained from the University Hospitals, Birmingham, UK (UHB) Trust Information Service and the Queen Elizabeth Hospital liver unit records on (a) the number of patients admitted to hospital because of paracetamol overdose and (b) the number of patients admitted to the liver unit with parcetamol-induced hepatotoxicity. RESULTS: Prior to legislation, an average of 360 people a year were admitted to UHB. However, following the change in legislation the number of admissions has fallen to an average of 250 people per year. This represents a reduction of 31%. Prior to legislation, an average of 76 people per year were admitted to the liver unit. Post-legislation this number has dropped to an average of 38 per year representing a reduction of 50%. CONCLUSION: Legislation restricting paracetamol pack-size reduced the incidence and severity of poisoning. PMID- 12911684 TI - The provision of pharmaceutical care in Denmark: a cross-sectional survey. AB - BACKGROUND: Pharmaceutical care was a concept initially defined in the early 1990s. It had its roots within clinical pharmacy, in the USA. In Denmark, pharmaceutical care has been part of the professional standards of practice for community pharmacy since 1995. OBJECTIVE: The objective of the present study was to investigate the provision of pharmaceutical care in community practice in Denmark. A focus of the study was the estimation of the frequency of medicine related problem identification and the process of problem management in the Danish pharmacies. METHODS: A cross-sectional questionnaire-based survey of all Danish community pharmacies was conducted (n = 288). The variables included: detection and identification of medicine-related problems, goal-setting for solving medicine-related problems, and documentation of efforts to solve these. The response rate was 75.7%. A non-respondent analysis was performed. RESULTS: On average three medicine-related problems per pharmacy were found within the working week prior to the survey. For two-thirds of those cases the type of problem involved was identified. For the other third, goals had been set to resolve the problem. Minimal documentation of these activities was reported. The primary collaborators in problem management were general practitioners and patients. CONCLUSION: Pharmaceutical care, in its fullest sense, as defined in policy documents in Denmark, was not evident in practice. While some aspects of pharmaceutical care were being performed, almost no documentation of efforts was taking place in community pharmacy. PMID- 12911685 TI - In vitro inhibitory effects of Kampo medicines on metabolic reactions catalyzed by human liver microsomes. AB - BACKGROUND: Although it is well known that drug-drug interactions may lead to toxicity and therapeutic failure, little is known about the incidence and consequences of herb-drug interactions in patients receiving Kampo medicines. METHODS: We evaluated the frequency of the combined use of Kampo medicines and Western drugs at Osaka University Hospital, and investigated the effects of these formulae on the metabolic activity of different cytochrome P450 (CYP) isoforms using pooled microsomes obtained from human liver. RESULTS: Twenty-two Kampo formulae were used together with 40 Western drugs catalyzed by the CYP isoforms CYP3A4, CYP2C9, CYP2D6 and CYP1A2. Among the Kampo medicines, HOCHUEKKI-TO, SHOSAIKO-TO, NINJINYOUEI-TO, SAIREI-TO and KAKKON-TO were most frequently used during the study period (1996-2000). These were co-administered with 11 categories of drugs, which are substrates for CYP3A4. HOCHUEKKI-TO and SAIREI-TO were competitive inhibitors of CYP3A4 with Ki values of 0.65 and 0.1 mg/mL, respectively. HOCHUEKKI-TO, SHOSAIKO-TO and SAIREI-TO inhibited the metabolic activities of CYP2C9, but had no effect on CYP2D6. HOCHUEKKI-TO and SAIREI-TO exhibited non-competitive inhibition of the metabolic activity of CYP2C9 with a similar Ki value (0.7-0.8 mg/mL). SAIRE-TO (0.25 mg/mL) was a potent inhibitor of CYP1A2 (inhibition > 68%). CONCLUSIONS: Frequently used Kampo medicines may interact with Western drugs, which are substrates for CYP3A4, CYP2C9 and CYP1A2. Their co-administration should be undertaken with care. PMID- 12911686 TI - Is oral azithromycin effective for the treatment of cyclosporine-induced gingival hyperplasia in cardiac transplant recipients? AB - Anecdotal evidence suggests that azithromycin is effective for the treatment of cyclosporine-induced gingival hyperplasia in solid-organ transplant recipients. We present the cases of two heart transplant patients who insidiously developed gingival hyperplasia, likely because of immunosuppression with cyclosporine, which was treated with azithromycin. Evidence supporting the efficacy of azithromycin in the treatment of cyclosporine-induced gingival hyperplasia in solid organ transplant recipients was searched for, identified, and then critically assessed. While no data were found specifically evaluating azithromycin in cardiac transplant patients, there were nine pertinent papers identified that evaluated the clinical question of interest in the renal transplant population [Wahlstrom et al. (1995) The New England Journal of Medicine 332, 753; Boran et al. (1996) Transplantation Proceedings 28, 2316; Gomez et al. (1997) Nephrology Dialysis Transplantation 12, 2694; Ljutic (1997) Dialysis & Transplantation 26, 787; Puig et al. (1997) Transplantation Proceedings 29, 2379; Nash et al. (1998) Transplantation 65, 1611; Nowicki et al. (1998) Annals of Transplantation 3, 25; Wirnsberger et al. (1998) Transplantation Proceedings 30, 2117; Citterio et al. (2001) Transplantation Proceedings 33, 2134]. These studies and case reports are summarized. While more evidence is required to support routine use of azithromycin for the treatment of cyclosporine induced gingival hyperplasia in cardiac transplant recipients, preliminary published evidence from renal transplant patients is certainly favourable. PMID- 12911687 TI - Use of pharmacoeconomics in prescribing research. Part 4: is cost-utility analysis a useful tool? AB - This paper is the fourth in the 'Research Note' series describing various aspects of pharmacoeconomics in prescribing research. This article describes cost-utility analysis, and how it can be incorporated into pharmacoeconomics. While this approach has intuitive appeal, the uncertainties about the methods for estimating utilities mean that the value of cost-utility analysis in prescribing research is still to be established. Importantly, cost-utility analysis may not capture aspects of quality of life that enable meaningful comparisons between prescribing choices at an individual or societal level. PMID- 12911688 TI - Novel antiviral agents: a medicinal plant perspective. AB - Several hundred plant and herb species that have potential as novel antiviral agents have been studied, with surprisingly little overlap. A wide variety of active phytochemicals, including the flavonoids, terpenoids, lignans, sulphides, polyphenolics, coumarins, saponins, furyl compounds, alkaloids, polyines, thiophenes, proteins and peptides have been identified. Some volatile essential oils of commonly used culinary herbs, spices and herbal teas have also exhibited a high level of antiviral activity. However, given the few classes of compounds investigated, most of the pharmacopoeia of compounds in medicinal plants with antiviral activity is still not known. Several of these phytochemicals have complementary and overlapping mechanisms of action, including antiviral effects by either inhibiting the formation of viral DNA or RNA or inhibiting the activity of viral reproduction. Assay methods to determine antiviral activity include multiple-arm trials, randomized crossover studies, and more compromised designs such as nonrandomized crossovers and pre- and post-treatment analyses. Methods are needed to link antiviral efficacy/potency- and laboratory-based research. Nevertheless, the relative success achieved recently using medicinal plant/herb extracts of various species that are capable of acting therapeutically in various viral infections has raised optimism about the future of phyto-antiviral agents. As this review illustrates, there are innumerable potentially useful medicinal plants and herbs waiting to be evaluated and exploited for therapeutic applications against genetically and functionally diverse viruses families such as Retroviridae, Hepadnaviridae and Herpesviridae PMID- 12911689 TI - Transfer of antibiotic resistance between commensal and pathogenic members of the Enterobacteriaceae under ileal conditions. AB - AIM: To determine the rate of antibiotic resistance transmission between commensal and pathogenic representatives of the Enterobacteriaceae. METHODS AND RESULTS: Through the use of a validated in vitro simulation of the porcine ileum, the transmission of antibiotic resistance was detected between commensal Escherichia coli, E. coli O157 and Salmonella spp. Countable transconjugant populations arose readily and, in one example, proved capable of indefinite persistence. CONCLUSIONS: Genetic material conferring antibiotic resistance is readily transmissible between members of the Enterobacteriaceae under ileal conditions. Recipient phenotype influences the persistence of multi-resistant transconjugants. SIGNIFICANCE AND IMPACT OF THE STUDY: The observation that the conjugal transmission of antibiotic resistance is commonplace under ileal conditions impacts primarily on the risk of food contamination by multi-resistant bacteria. The establishment of a multi-resistant transconjugant population as a dominant member of the microflora maintains a genetic reservoir of antimicrobial resistance. PMID- 12911690 TI - Metabiosis of proteolytic moulds and Salmonella in raw, ripe tomatoes. AB - AIMS: The aim of this study was to determine the survival and growth characteristics of Salmonella enterica in sound and chill-injured tomatoes as influenced by co-infection with proteolytic moulds. METHODS AND RESULTS: Sound (not chill injured) raw, ripe tomatoes (Lycopersicon esculentum Mill. 'Roma') were inoculated with a five-serotype mixture of S. enterica and/or Alternata alternata (two strains), Cladosporium herbarum and C. cladosporioides. Simultaneous and delayed (3 days) inoculation of tomatoes with Salmonella and each mould was studied. Growth of moulds in sound tomatoes stored at 15 and 25 degrees C for up to 10 days was accompanied by increased pH of radial pericarp tissue (pulp), which enhanced the growth of Salmonella. Growth of moulds and Salmonella at 25 degrees C was enhanced in chill-injured tomatoes compared with sound tomatoes. CONCLUSIONS: Growth of proteolytic moulds in tomatoes stored at conditions simulating those commonly used in commercial postharvest storage and handling promotes the growth of Salmonella that may be an incidental contaminant. SIGNIFICANCE AND IMPACT OF THE STUDY: Discarding tomatoes that are infected by moulds is important in handling and minimal processing practices designed to minimize the risk of human salmonellosis. PMID- 12911691 TI - The role of visible faecal material as a vehicle for generic Escherichia coli, coliform, and other enterobacteria contaminating poultry carcasses during slaughtering. AB - AIMS: A comparison of Enterobacteriaceae, coliform and Escherichia coli counts in chicken carcasses with and without visible faecal contamination was conducted to evaluate the role of contamination as a vehicle for generic E. coli, coliform and other enterobacteria contaminating broiler chicken carcasses when processed under routine commercial operations. METHODS AND RESULTS: Samples were removed from the processing line immediately after evisceration, inside-outside shower and chilling for microbiological analysis. After evisceration, mean counts were significantly different only for E. coli (P < or = 0.05) in chicken carcasses with and without visible faecal contamination. While the spray wash practice was not efficient enough for complete removal of the visible contamination from carcasses, leading to microbiological reduction percentages lower than expected, 25 ppm chlorinated water chilling did reduce the contamination level considerably in all samples. CONCLUSIONS: Carcasses with and without visible faecal contamination harboured E. coli and other potentially hazardous enterobacteria. E. coli was the predominant strain isolated in all samples, Enterobacter cloacae being next most frequent. SIGNIFICANCE AND IMPACT OF THE STUDY: The zero tolerance of visible faecal contamination requirement alone is not sufficient to assure safety and to improve the microbial quality of carcasses. PMID- 12911692 TI - Flocculation and cell surface characterization of Kloeckera apiculata from wine. AB - AIMS: To characterize and analyze the flocculation phenomenon of Kloeckera apiculata mc1 from Argentinian wine to understand the cell-cell interaction pattern. METHODS AND RESULTS: Kloeckera apiculata mc1 possess intense cell-cell interactions in MYPG medium (0.5% malt extract, 1% yeast extract, 2% glucose, 2% peptone), pH 5.5 by shaking at 25 degrees C. Optimum flocculation is observed at pH 4.5 in the presence of 3 mmol l-1 Ca2+. The flocculation is induced by peptone and malt extract and not by yeast extract and is reversed by 50 mmol l-1 galactose or lactose. The flocculation is highly susceptible to pronase, chymotrypsine and proteases types IV and XXVII and is partially resistant to trypsin. The electronic microscopy shows that the cells are attached to each other along their sides by fine hair-like threads. CONCLUSIONS: The mechanism of flocculation of K. apiculata mc1 is mediated by protein-carbohydrate interaction, stabilized by Ca2+. SIGNIFICANCE AND IMPACT OF THE STUDY: The use of selected pure yeast inocula of known ability is preferred to wine elaboration, therefore the indigenous flora must be avoided and the flocculation of K. apiculata could be an economic method to do it. PMID- 12911693 TI - Biodiversity in Lactobacillus helveticus strains present in natural whey starter used for Parmigiano Reggiano cheese. AB - AIMS: Lactobacillus helveticus is the dominant microflora of the natural whey starters used for Parmigiano Reggiano cheese making. The aim of this work was to study the biodiversity of different strains of Lact. helveticus present in six cultures and to compare them with strains of the same species previously isolated from natural whey cultures used for Grana Padano and Provolone cheeses. METHODS AND RESULTS: Twenty different biotypes of Lact. helveticus strains were identified combining the results deriving from SDS-PAGE of cell surface proteins and PCR fingerprinting using M13 as a primer. The biotypes were present in varying amounts in the six natural whey starters and the biodiversity was demonstrated not only within the whey cultures, but also between the whey cultures. CONCLUSIONS: Lact. helveticus strains isolated from Parmigiano Reggiano whey cultures analysed by PCR M13, SDS-PAGE and RFLP were distinguishable from Lact. helveticus strains of different dairy origin, namely Grana Padano and Provolone natural whey starters. SIGNIFICANCE AND IMPACT OF THE STUDY: The presence of different Lact. helveticus biotypes seems to be related to the specific ecosystem of cheese making and may be considered as one of the elements contributing to the typicality of Parmigiano Reggiano cheese. PMID- 12911694 TI - Inhibition of Staphylococcus aureus by the commensal bacteria of human milk. AB - AIMS: To study the bacterial diversity in expressed human milk with a focus on detecting bacteria with an antimicrobial activity against Staphylococcus aureus, known as a causative agent of maternal breast infections and neonatal infections. METHODS AND RESULTS: Random isolates (n = 509) were collected from breast milk samples (n = 40) of healthy lactating women, genotypically identified, and tested for antimicrobial activity against Staph. aureus. Commensal staphylococci (64%) and oral streptococci (30%), with Staph. epidermidis, Strep. salivarius, and Strep. mitis as the most frequent isolates, were the predominant bacterial species in breast milk. One-fifth of Staph. epidermidis and half of Strep. salivarius isolates suppressed growth of Staph. aureus. Enterococci (Ent. faecalis), isolated from 7.5% of samples, and lactic acid bacteria (LAB) (Lactobacillus rhamnosus, Lact. crispatus, Lactococcus lactis, Leuconoctoc mesenteroides), isolated from 12.5% of samples, were also effective against Staph. aureus. One L. lactis isolate was shown to produce nisin, a bacteriocin used in food industry to prevent bacterial pathogens and spoilage. CONCLUSIONS: Expressed breast milk contains commensal bacteria, which inhibit Staph. aureus. SIGNIFICANCE AND IMPACT OF THE STUDY: The strains inhibitory against the pathogen Staph. aureus have potential use as bacteriotherapeutic agents in preventing neonatal and maternal breast infections caused by this bacterium. PMID- 12911695 TI - Acclimation and repair of DNA damage in recombinant bioluminescent Escherichia coli. AB - AIMS: The aim of this study is to understand different adaptive responses in bacteria caused by three different mutagens, namely, an intercalating agent, an alkylating agent and a hydroxylating agent, and the repair systems according to the type of DNA damage, that is, DNA cross-linking and delayed DNA synthesis, alkylation and hydroxylation of DNA. A recombinant bioluminescent Escherichia coli, DPD2794 with the recA promoter fused to luxCDABE originating from Vibrio fischeri, was used in this study. METHODS AND RESULTS: The recombinant bioluminescent E. coli strain DPD2794, containing a recA promoter fused to luxCDABE from V. fischeri, was used to detect adaptive and repair responses to DNA damage caused by mitomycin C (MMC), and these responses were compared with those when the cells were induced with N-methyl-N-nitro-N-nitrosoguanidine (MNNG) and hydrogen peroxide (H2O2). The response ratio between the induced samples and that of the controls decreased suddenly when the induced culture was used in further inductions, indicating a possible adaptive response to DNA damage. DNA damage, or the proteins produced, because of MMC addition does not appear to be completely resolved until the seventh sub-culture after the initial induction, whereas simple damage, such as the base modification caused by MNNG and H2O2, appears to be repaired rapidly as evidenced by the quick recovery of sensitivity. CONCLUSIONS: These results suggest that it takes more time to completely repair DNA damage caused by MMC, as compared with a simple repair such as that required for the damage caused by MNNG and H2O2. Therefore, repair of the damage caused by these three mutagens is controlled by different regulons, even though they all induced the recA promoter. SIGNIFICANCE AND IMPACT OF THE STUDY: Using a bioluminescent E. coli harbouring a recA promoter-lux fusion, it was found that different adaptive responses and repair systems for DNA damage caused by several mutagens exists in E. coli. PMID- 12911696 TI - Effect of trehalose on survival of Bradyrhizobium japonicum during desiccation. AB - AIMS: A major reason for the ineffectiveness of legume inoculants in the field is the rapid death of rhizobia because of desiccation. The major purpose of this study was to identify conditions under which alpha,alpha-trehalose would improve survival of Bradyrhizobium japonicum during desiccation. METHODS AND RESULTS: Trehalose was added to cultures just prior to desiccation or was supplied to bacteria during the 6-day growth period. A wide variety of trehalose concentrations was tested. Trehalose added to cultures at the time of desiccation improved survival slightly, but trehalose loading during growth was much more effective in protection against desiccation. Growth of bacteria with 3 mmol l-1 trehalose increased trehalose concentration in cells by about threefold and increased survival of cells placed on soya bean [Glycine max (L.) Merr.] seeds by two- to four-fold after 2 or 24 h. Average of overall results indicate that growth of bacteria with trehalose in the medium resulted in a 294% increase in survival after 24 h of desiccation. The concentration of trehalose in cells was very highly correlated with survival of bacteria. When trehalose-loaded cells were suspended in buffer or water, 60-85% of cellular trehalose was lost in about 1 h and, in spite of these losses, survival during desiccation was not reduced. CONCLUSIONS: Accumulation of trehalose in the cytoplasm is critical to the survival of B. japonicum during desiccation. Increasing the periplasmic concentration of trehalose is also beneficial but is not so critical as the concentration of trehalose in the cytoplasm. Because B. japonicum cannot utilize trehalose as a carbon source, cells can be loaded with trehalose by providing the disaccharide during the growth period. SIGNIFICANCE AND IMPACT OF THE STUDY: Although it may not be practical to use trehalose as a carbon source in inoculant production, it may be possible to engineer greater trehalose accumulation in rhizobia. Trehalose concentration in cells should be a useful predictor of survival during desiccation. PMID- 12911697 TI - Mediation of arsenic oxidation by Thiomonas sp. in acid-mine drainage (Carnoules, France). AB - AIMS: To isolate, identify, and characterize heterotrophic bacteria in acid-mine drainage that mediate oxidation of As(III). METHODS AND RESULTS: Samples of acid mine drainage were collected over a period of 14 months. Heterotrophic and non obligatory acidophilic bacteria in the samples were cultured on a solid medium (pH 7.0-7.2), and three strains were isolated. The three different strains belong to the genus Thiomonas, and have more than 99% homology with the group Ynys1. Culturing in mineral media demonstrated that the isolated strains used thiosulphate as an energy source, and oxidized iron in the presence of thiosulphate. However, none of the strains were able to oxidize arsenic in the presence of thiosulphate, nor could they use iron or arsenic alone as an energy source. In vitro experiments demonstrated that two of the Thiomonas strains were able to oxidize more than 90% of the As(III) present in the acid-mine drainage, whereas no abiotic oxidation of arsenic occurred. CONCLUSIONS: Two strains of newly identified Thiomonas sp. found in acid-mine drainage are capable of oxidizing arsenic. SIGNIFICANCE AND IMPACT OF STUDY: These results represent the first reported oxidation of arsenic by Thiomonas sp. Biologically mediated oxidation and subsequent immobilization of arsenic is of great interest for the remediation of contaminated mine sites. PMID- 12911698 TI - A model for the thermal inactivation of micro-organisms. AB - AIMS: To mathematically model published thermal inactivation data sets using an empirical model based on a double Arrhenius function. METHODS AND RESULTS: A mathematical model, the log R-fat, provided an excellent description of the data sets available: the thermal inactivation of Salmonella anatum at 55 degrees C, Pseudomonas viscosa at 48 degrees C and Streptococcus faecalis at 60 degrees C; Clostridium botulinum spores at various temperatures in the range of 101-121 degrees C (two data sets); thermal inactivation of Salmonella Bedford over the temperature range 50-58 degrees C, water activity range of 0.94-0.99 and a pH range of 4-7; Bacillus stearothermophilus spores from 105 to 121 degrees C and the dry heat sterilization of an indigenous mesophilic soil population over the temperature range of 120-160 degrees C. CONCLUSIONS: The log R-fat model, derived from previously published chemical inactivation studies provides as good, if not better, description of thermal inactivation kinetics as other published models. The model does not invoke either of the two hypotheses of inactivation: the mechanistic or vitalistic, although it is closely linked to descriptions of the former. SIGNIFICANCE AND IMPACT OF THE STUDY: The log R-fat double Arrhenius function provides the investigator with a relatively simple and easy mathematical model to apply to data of thermal inactivation. This model may allow a more accurate description of thermal food processing, especially when the safety of marginal heat processes are concerned. PMID- 12911699 TI - Molecular characterization of the microbial species that colonize human ileal and colonic mucosa by using 16S rDNA sequence analysis. AB - AIM: The aim of this study was to characterize the bacterial community adhering to the mucosa of the terminal ileum, and proximal and distal colon of the human digestive tract. METHODS AND RESULTS: Pinch samples of the terminal ileum, proximal and distal colon were taken from a healthy 35-year-old, and a 68-year old subject with mild diverticulosis. The 16S rDNA genes were amplified using a low number of PCR cycles, cloned, and sequenced. In total, 361 sequences were obtained comprising 70 operational taxonomic units (OTU), with a calculated coverage of 82.6%. Twenty-three per cent of OTU were common to the terminal ileum, proximal colon and distal colon, but 14% OTU were only found in the terminal ileum, and 43% were only associated with the proximal or distal colon. The most frequently represented clones were from the Clostridium group XIVa (24.7%), and the Bacteroidetes (Cytophaga-Flavobacteria-Bacteroides) cluster (27.7%). CONCLUSION: Comparison of 16S rDNA clone libraries of the hindgut across mammalian species confirms that the distribution of phylogenetic groups is similar irrespective of the host species. Lesser site-related differences within groups or clusters of organisms, are probable. SIGNIFICANCE AND IMPACT: This study provides further evidence of the distribution of the bacteria on the mucosal surfaces of the human hindgut. Data contribute to the benchmarking of the microbial composition of the human digestive tract. PMID- 12911700 TI - Substrate induction of isomaltulose synthase in a newly isolated Klebsiella sp. LX3. AB - AIMS: Production of isomaltulose by newly isolated Klebsiella sp. LX3. METHODS AND RESULTS: The bacterial isolate LX3, which transforms sucrose to isomaltulose and trehalulose, has been isolated from a soil sample in Singapore. Morphological and biochemical analysis, as well as 16s rRNA sequence demonstrated that the isolate could represent a new member of genus Klebsiella. The strain has several interesting features. The immobilized cells of Klebsiella sp. LX3 convert more than 99% of sucrose to products that consist of more than 87% of isomaltulose, 11.6% of trehalulose, and <1% of glucose. CONCLUSIONS: The production of isomaltulose synthase in isolate LX3 is inducible by its substrate sucrose and the sugars containing a fructofuranosyl group. SIGNIFICANCE AND IMPACT OF STUDY: It would be useful for future biotechnological applications to understand the structural features or motifs of the isomaltulose synthases that determine the sucrose conversion efficiency and the ratio of the conversion products. PMID- 12911701 TI - Effect of some abiotic factors on the biological activity of Gluconacetobacter diazotrophicus. AB - AIMS: The effect of some abiotic factors, dryness, heat and salinity on the growth and biological activity of Gluconacetobacter diazotrophicus, and the influence of a salt stress on some enzymes involved in carbon metabolism of these bacteria is studied under laboratory conditions. METHODS AND RESULTS: Strain PAL 5 of G. diazotrophicus was incubated under different conditions of drying, heat and salinity. Cells showed tolerance to heat treatments and salt concentrations, and sensitivity to drying conditions. Higher NaCl dosage of 150 and 200 mmol l -1 limited its growth and drastically affected the nitrogenase activity and the enzymes glucose dehydrogenase, alcohol dehydrogenase, fumarase, isocitrate dehydrogenase and malate dehydrogenase. CONCLUSIONS: Gluconacetobacter diazotrophicus, despite its endophytic nature, tolerated heat treatments and salinity stress, but its nitrogenase activity and carbon metabolism enzymes were affected by high NaCl dosage. SIGNIFICANCE AND IMPACT OF THE STUDY: The investigation of the biological activity of G. diazotrophicus in response to different abiotic factors led to more knowledge of this endophyte and may help to clarify pathways involved in its transmission into the host plant. PMID- 12911702 TI - Influence of groundwater characteristics on the survival of enteric viruses. AB - AIMS: This study was undertaken to further understand the processes affecting the persistence of enteric viruses in groundwater. METHODS AND RESULTS: Varying temperature, oxygen and nutrient levels were tested in the presence and absence of groundwater micro-organisms to determine which of the factors tested had dominant influence on the decay of Escherichia coli, the bacteriophage MS2, poliovirus and coxsackievirus. The results indicated that the most influential factor affecting the decay of the viruses and E. coli was the presence of groundwater micro-organisms. The results also implied that temperature, the presence of oxygen and nutrient levels indirectly influence viruses and E. coli decay by influencing the activity of the groundwater micro-organisms. CONCLUSIONS: E. coli and the viruses displayed maximum decay under aerobic conditions, at 28 degrees C without the addition of nutrients in the presence of groundwater micro-organisms. SIGNIFICANCE AND IMPACT OF THE STUDY: The results suggest that if the mode of action of the groundwater micro-organisms could be determined then the decay of viral pathogens in recharged waters may be more easily predicted. PMID- 12911703 TI - Effect of fermented soya beans on diarrhoea and feed efficiency in weaned piglets. AB - AIMS: To evaluate anti-diarrhoeal and growth enhancing properties of fermented soya beans in weaned piglets. METHODS AND RESULTS: In a first phase piglet diet, toasted full-fat soya beans (20%) were replaced with either cooked soya beans or Rhizopus microsporus or Bacillus subtilis fermented soya beans. The effect on the incidence, severity and duration of diarrhoea in enterotoxigenic Escherichia coli (ETEC)-challenged weaned piglets was determined (pen trial, 24 piglets per treatment). Severity of diarrhoea was significantly less on the diet with Rhizopus-fermented soya beans compared with the control diet containing toasted soya beans. Piglets fed fermented soya beans showed increased feed intake (13 and 12%), average daily weight gain (18 and 21%) and feed efficiency (3 and 8%) (for Rhizopus and Bacillus-fermented soya beans, respectively). However, in the treatment groups an unequal mortality and a potential unequal distribution of receptor-positive piglets were observed. CONCLUSIONS: Cooked and fermented soya beans could be beneficial in the control of diarrhoea in ETEC-challenged weaned piglets (particularly Rhizopus fermented) and significantly improved weight gain and feed intake (particularly Bacillus fermented). SIGNIFICANCE AND IMPACT OF THE STUDY: Fermented soya beans could offer benefits with respect to the control of diarrhoea and feed efficiency in piglets. PMID- 12911704 TI - The use of amplified flanking region-PCR in the isolation of laccase promoter sequences from the edible fungus Pleurotus sajor-caju. AB - AIMS: To determine the regulation of laccase isozyme gene transcription in Pleurotus sajor-caju in response to different aromatic inducers and physiological parameters. METHODS AND RESULTS: The promoter regions for each of four different laccase isozymes were cloned from P. sajor-caju, using amplified flanking region PCR (AFR-PCR). Sequences stretching 724, 214, 840 and 1740 bp upstream from the predicted start codons for lac1, lac2, lac3 and lac4, respectively, were cloned in each case and analysed for the presence of putative transcriptional response elements. A number of putative response elements including metal response elements, xenobiotic response elements and antioxidant response elements appear to be present. In addition putative consensus sequences such as those for the binding of AP1, AP2, creA and NIT2 transcription factors, which are involved in nitrogen and carbon regulation in different fungi, are also present in the promoter regions of some of the isozymes. CONCLUSIONS: These elements may be involved in the transcriptional regulation of laccase gene expression in P. sajor caju. SIGNIFICANCE AND IMPACT OF THE STUDY: The presence of a number of putative transcriptional response elements in the promoter regions of different isozyme genes indicates a potential role for these sites in regulating laccase gene transcription in P. sajor-caju. In addition this work demonstrates the potential usefulness of AFR-PCR as a technique to clone fungal DNA sequences located upstream from known sequences. PMID- 12911705 TI - Evaluation of the pH-dependent, stationary-phase acid tolerance in Listeria monocytogenes and Salmonella Typhimurium DT104 induced by culturing in media with 1% glucose: a comparative study with Escherichia coli O157:H7. AB - AIMS: To comparatively evaluate the adaptive stationary-phase acid tolerance response (ATR) in food-borne pathogens induced by culturing in glucose-containing media, as affected by strain variability and antibiotic resistance, growth temperature, challenge pH and type of acidulant. METHODS AND RESULTS: Antibiotic resistant or sensitive strains of Listeria monocytogenes, Salmonella including S. Typhimurium DT104, and Escherichia coli O157:H7 were cultured (30 degrees C for 24 h; 10 degrees C for up to 14 days) in trypticase soya broth with yeast extract (TSBYE) with 1% or without glucose to induce or prevent acid adaptation, respectively. Cultures were subsequently exposed to pH 3.5 or 3.7 with lactic or acetic acid at 25 degrees C for 120 min. Acid-adapted cultures were more acid tolerant than nonadapted cultures, particularly those of L. monocytogenes and Salmonella. No consistent, positive or negative, influence of antibiotic resistance on the pH-inducible ATR or acid resistance (AR) was observed. Compared with 30 degrees C cultures, growth and acid adaptation of L. monocytogenes and S. Typhimurium DT104 at 10 degrees C markedly reduced their ATR and AR in stationary phase. E. coli O157:H7 had the greatest AR, relying less on acid adaptation. A 0.2 unit difference in challenge pH (3.5-3.7) caused great variations in survival of acid-adapted and nonadapted cells. CONCLUSIONS: Culturing L. monocytogenes and Salmonella to stationary phase in media with 1% glucose induces a pH-dependent ATR and enhances their survival to organic acids; thus, this method is suitable for producing acid-adapted cultures for use in food challenge studies. SIGNIFICANCE AND IMPACT OF THE STUDY: Bacterial pathogens may become acid-adapted in foods containing glucose or other fermentable carbohydrates. Low storage temperatures may substantially decrease the stationary-phase ATR of L. monocytogenes and S. Typhimurium DT104, but their effect on ATR of E. coli O157:H7 appears to be far less dramatic. PMID- 12911706 TI - Nitrosative stress induced cytotoxicity in Giardia intestinalis. AB - AIMS: To investigate the antigiardial properties of the nitrosating agents: sodium nitrite, sodium nitroprusside and Roussin's black salt. METHODS AND RESULTS: Use of confocal laser scanning microscopy and flow cytometry indicated permeabilization of the plasma membrane to the anionic fluorophore, DiBAC4(3) [bis(1,3-dibutylbarbituric acid) trimethine oxonol]. Loss of plasma membrane electrochemical potential was accompanied by loss of regulated cellular volume control. Changes in ultrastructure revealed by electron microscopy and capacity for oxygen consumption, were also consequences of nitrosative stress. Roussin's black salt (RBS), active at micromolar concentrations was the most potent of the three agents tested. CONCLUSIONS: These multitargeted cytotoxic agents affected plasma membrane functions, inhibited cellular functions in Giardia intestinalis and led to loss of viability. SIGNIFICANCE AND IMPACT OF THE STUDY: Nitrosative damage, as an antigiardial strategy, may have implications for development of chemotherapy along with suggesting natural host defence mechanisms. PMID- 12911707 TI - Stability of beta-carotene in spray dried preparation of Rhodotorula glutinis mutant 32. AB - AIMS: To obtain beta-carotene-rich dry cell preparation from mutant 32 of Rhodotorula glutinis and determination of its pigment stability. METHODS AND RESULTS: The mutant 32 of R. glutinis was grown in a 14 l stirred tank fermenter. Cell mass was concentrated 10-fold by cross-flow microfiltration and then spray dried. Butylated hydroxy toluene (BHT) and d-tocopherol were used as protecting agents. A two-level, three-variable, factorial optimization was performed to achieve moisture-free, non-viable and beta-carotene-rich feed additive. CONCLUSIONS: The beta-carotene and cell mass in stirred tank fermenter were found to be 54 +/- 5 mg l-1 and 12.8 +/- 2 g l-1, respectively. In the presence of BHT, 97 +/- 3% (w/w) beta-carotene was recovered for all the inlet temperatures studied. The best beta-carotene and yeast powder recoveries were obtained at 160 degrees C, 11.6% (w/v) cell mass concentration and 1 g l-1 BHT. The pigments inside dried yeast powder were stable in dark and cold condition for at least 10 weeks. The purified beta-carotene got almost totally denatured, under similar conditions of storage, within 76 h. SIGNIFICANCE AND IMPACT OF THE STUDY: Spray dried and stable preparation of beta-carotene-rich yeast, R. glutinis can provide alternative source of beta-carotene for use in animal nutrition. PMID- 12911708 TI - Formation of natural biofilms during chlorine dioxide and u.v. disinfection in a public drinking water distribution system. AB - AIMS: The influence of two disinfection techniques on natural biofilm development during drinking water treatment and subsequent distribution is compared with regard to the supply of a high-quality drinking water. METHODS AND RESULTS: The growth of biofilms was studied using the biofilm device technique in a real public technical drinking water asset. Different pipe materials which are commonly used in drinking water facilities (hardened polyethylene, polyvinyl chloride, steel and copper) were used as substrates for biofilm formation. Apart from young biofilms, several months old biofilms were compared in terms of material dependence, biomass and physiological state. Vital staining of biofilms with 5-cyano-2,3-ditolyl tetrazolium chloride (CTC) and the DNA-specific 4',6 diamidino-2-phenylindole (DAPI) staining resulted in a significant difference in physiological behaviour of biofilm populations depending on the disinfection technique. Compared with chlorine dioxide disinfection (0.12-0.16 mg l-1), the respiratory activities of the micro-organisms were increased on all materials during u.v. disinfection (u.v.254; 400 J m-2). The biofilm biocoenosis was analysed by in situ hybridization with labelled oligonucleotides specific for some subclasses of Proteobacteria. Using PCR and additional hybridization techniques, the biofilms were also tested for the presence of Legionella spp., atypical mycobacteria and enterococci. The results of the molecular-biological experiments in combination with cultivation tests showed that enterococci were able to pass the u.v. disinfection barrier and persist in biofilms of the distribution system, but not after chlorine dioxide disinfection. CONCLUSIONS: The results indicated that bacteria are able to regenerate and proliferate more effectively after u.v. irradiation at the waterworks, and chlorine dioxide disinfection appears to be more applicative to maintain a biological stable drinking water. SIGNIFICANCE AND IMPACT OF THE STUDY: As far as the application of u.v. disinfection is used for conditioning of critical water sources for drinking water, the efficiency of u.v. irradiation in natural systems should reach a high standard to avoid adverse impacts on human health. PMID- 12911709 TI - Isolation of partial toxR gene of Vibrio harveyi and design of toxR-targeted PCR primers for species detection. AB - AIMS: To differentiate Vibrio harveyi from closely related Vibrio species by toxR sequence analysis and design primers for the specific detection of the shellfish pathogen. METHODS AND RESULTS: The partial toxR homologue from the shellfish pathogen V. harveyi was isolated by PCR using degenerate primers. The 578-bp toxR fragment from V. harveyi, that exhibited highest homology with partial toxR of V. parahaemolyticus (68%), is predicted to encode for a polypeptide with 192 amino acid residues. Alignment of the V. harveyi toxR nucleotide and deduced amino acid sequence with those from other Vibrio species revealed the presence of the fairly characteristic conserved transcription activation and transmembrane domain as well as the divergent membrane tether region that may be targeted for the development of species-specific oligonucleotide primers. Consequently, PCR primers that could amplify a 390-bp gene fragment in V. harveyi were designed by targeting portions of the V. harveyi toxR that display variability with toxR sequences from other Vibrio species. The 390-bp-amplicon was detected in all V. harveyi strains examined except in the nontarget bacteria and unexpectedly, in two shrimp-derived strains (VIB 391 and STD 3-101) from Thailand and Ecuador. Results show that strains exhibiting the 390-bp amplicon mostly belong to the same cluster based on previous amplified fragment length polymorphism data while strains which were previously unclustered or unclassified did not display the 390 bp PCR product. CONCLUSIONS: The toxR sequence variation could differentiate V. harveyi from closely related Vibrio species. A PCR protocol amplifying a 390-bp fragment of the V. harveyi toxR was established and could be useful in the specific and rapid detection of the species. SIGNIFICANCE AND IMPACT OF THE STUDY: The molecular approaches reported in this study could facilitate the early diagnosis and surveillance of luminous vibriosis in hatchery-reared fish and shellfish species through rapid identification and specific detection of causal agent. PMID- 12911710 TI - Analysis of environmental Escherichia coli isolates for virulence genes using the TaqMan PCR system. AB - AIMS: To assess the presence of virulence genes in environmental and foodborne Escherichia coli isolates using the TaqMan PCR system. METHODS AND RESULTS: Three TaqMan pathogen detection kits called O157:H7, StxI and StxII were used to investigate the presence of virulence genes in Escherichia coli isolates. All 54 foodborne E. coli O157:H7 isolates showed expected results using these kits. Ninety (15%) of 604 environmental isolates gave positive amplification with an O157:H7-specific kit. TaqMan PCR amplification products from these 90 isolates were analysed by agarose gel electrophoresis, and 90% (81 of 90) of the environmental samples contained the expected PCR product. Sixty-six of these 90 were chosen for serotyping tests and only 35% (23 of 66) showed agglutination with both anti-O157 and anti-H7 antibodies. Further ribotyping of 16 sero positive isolates in an automated Riboprinter did not identify these to be O157:H7. Multiplex PCR with primers for eaeA, stxI and stxII genes was used to confirm the TaqMan results in 10 selected environmental isolates. CONCLUSIONS: All three TaqMan pathogen detection kits were useful for virulence gene analysis of prescreened foodborne O157:H7 isolates, while the O157:H7-specific kit may not be suitable for virulence gene analysis of environmental E. coli isolates, because of high false positive identification. SIGNIFICANCE AND IMPACT OF THE STUDY: The ability to rapidly identify the presence of pathogenic E. coli in food or environmental samples is essential to avert outbreaks. These results are of importance to microbiologists seeking to use TaqMan PCR to rapidly identify pathogenic E. coli in environmental samples. Furthermore, serotyping may not be a reliable method for identification of O157:H7 strains. PMID- 12911711 TI - Establishing populations of Megasphaera elsdenii YE 34 and Butyrivibrio fibrisolvens YE 44 in the rumen of cattle fed high grain diets. AB - AIM: To determine whether Megasphaera elsdenii YE34 (lactic acid degrader) and Butyrivibrio fibrisolvens YE44 (alternative starch utilizer to Streptococcus bovis) establish viable populations in the rumen of beef cattle rapidly changed from a forage-based to a grain-based diet. METHODS AND RESULTS: Five steers were inoculated with the two bacterial strains (YE34 and YE44) and five served as uninoculated controls. With the exception of one animal in the control group, which developed acidosis, all steers rapidly adapted to the grain-based diet without signs of acidosis (pH decline and accumulation of lactic acid). Bacterial populations of S. bovis, B. fibrisolvens and M. elsdenii were enumerated using real-time Taq nuclease assays. Populations of S. bovis remained constant (except in the acidotic animal) at ca 10(7) cell equivalents (CE) ml-1 throughout the study. Megasphaera elsdenii YE34, was not detectable in animals without grain in the diet, but immediately established in inoculated animals, at 10(6) CE ml-1, and increased 100-fold in the first 4 days following inoculation. Butyrivibrio fibrisolvens, initially present at 10(8) CE ml-1, declined rapidly with the introduction of grain into the diet and was not detectable 8 days after grain introduction. CONCLUSION: Megasphaera elsdenii rapidly establishes a lactic acid utilizing bacterial population in the rumen of grain-fed cattle 7-10 days earlier than in uninoculated cattle. SIGNIFICANCE AND IMPACT OF THE STUDY: The study has demonstrated that rumen bacterial populations, and in particular the establishment of bacteria inoculated into the rumen for probiotic use, can be monitored by real-time PCR. PMID- 12911712 TI - Survival of Escherichia coli O157:H7, Listeria monocytogenes 4b and Yersinia enterocolitica O3 in different yogurt and kefir combinations as prefermentation contaminant. AB - AIMS: To compare microbiological safety of yogurt, kefir and different combinations of yogurt and kefir samples by using three foodborne pathogenic strains (Escherichia coli O157:H7, Listeria monocytogenes 4b and Yersinia enterocolitica O3) as indicators. METHODS AND RESULTS: Fresh yogurt and kefir drinks were added to pasteurized milk at a 5% rate either separately or together, and then incubated at different temperatures (43 degrees C for yogurt and 30 degrees C for kefir), depending on appropriate growth temperature of their starter microflora. While traditional yogurt was found to be the least suppressive on the three pathogenic micro-organisms, samples obtained from two subsequent fermentation process (samples fermented at 43 degrees C for 3 h and at 30 degrees C for 21 h) were more suppressive than that of traditional kefir. There was no significant survival difference between E. coli O157:H7 and L. monocytogenes 4b in samples tested (P > 0.05), but Y. enterocolitica O3 was more susceptible than other two test strains (P < 0.05). CONCLUSIONS: The microbiological safety of the dairy product fermented at two consecutive periods was superior than that of traditional yogurt or kefir alone. SIGNIFICANCE AND IMPACT OF THE STUDY: These experiments may mimic what happens when yogurt and kefir starter micro-organisms are combined in a milk fermentation process with different time and temperature periods. PMID- 12911713 TI - Germination of spores of Bacillus subtilis with dodecylamine. AB - AIMS: To determine the properties of Bacillus subtilis spores germinated with the alkylamine dodecylamine, and the mechanism of dodecylamine-induced spore germination. METHODS AND RESULTS: Spores of B. subtilis prepared in liquid medium were germinated efficiently by dodecylamine, while spores prepared on solid medium germinated more poorly with this agent. Dodecylamine germination of spores was accompanied by release of almost all spore dipicolinic acid (DPA), degradation of the spore's peptidoglycan cortex, release of the spore's pool of free adenine nucleotides and the killing of the spores. The dodecylamine germinated spores did not initiate metabolism, did not degrade their pool of small, acid-soluble spore proteins efficiently and had a significantly lower level of core water than did spores germinated by nutrients. As measured by DPA release, dodecylamine readily induced germination of B. subtilis spores that: (a) were decoated, (b) lacked all the receptors for nutrient germinants, (c) lacked both the lytic enzymes either of which is essential for cortex degradation, or (d) had a cortex that could not be attacked by the spore's cortex-lytic enzymes. The DNA in dodecylamine-germinated wild-type spores was readily stained, while the DNA in dodecylamine-germinated spores of strains that were incapable of spore cortex degradation was not. These latter germinated spores also did not release their pool of free adenine nucleotides. CONCLUSIONS: These results indicate that: (a) the spore preparation method is very important in determining the rate of spore germination with dodecylamine, (b) wild-type spores germinated by dodecylamine progress only part way through the germination process, (c) dodecylamine may trigger spore germination by a novel mechanism involving the activation of neither the spore's nutrient germinant receptors nor the cortex lytic enzymes, and (d) dodecylamine may trigger spore germination by directly or indirectly activating release of DPA from the spore core, through the opening of channels for DPA in the spore's inner membrane. SIGNIFICANCE AND IMPACT OF THE STUDY: These results provide new insight into the mechanism of spore germination with the cationic surfactant dodecylamine, and also into the mechanism of spore germination in general. New knowledge of mechanisms to stimulate spore germination may have applied utility, as germinated spores are much more sensitive to processing treatments than are dormant spores. PMID- 12911714 TI - Systematic review of adjuvant therapy for early stage (epithelial) ovarian cancer. AB - A systematic review and meta analysis has been undertaken in order to evaluate the effectiveness of adjuvant therapy following surgery for early ovarian cancer. Trials reported since 1990 have been of a higher quality enabling a meta analysis of adjuvant chemotherapy vs adjuvant radiotherapy and a meta analysis of adjuvant chemotherapy vs observation. There was no significant difference between radiotherapy and chemotherapy, though these comprised studies which demonstrated considerable heterogeneity. Chemotherapy did confer significant benefit over observation in terms of both overall and disease free survival. Except for women in whom adequate surgical staging has revealed well differentiated disease confined to one or both ovaries with intact capsule, platinum chemotherapy should be offered to reduce risk of recurrence. PMID- 12911715 TI - Rodent models for ovarian cancer research. AB - Animal models that are biologically and clinically relevant are essential for conducting research to investigate the pathophysiologic progression of disease and to develop diagnostic or therapeutic strategies. Several rodent models that vary in methods of induction allow appropriate in vivo evaluation for ovarian cancer. The types of rodent models discussed include chemically (nonhormonal and hormonal) induced, genetic (knockout and transgenic), xenograft, and syngeneic. A summary of the available rodent models is provided with a discussion of the advantages and disadvantages of each. Optimization and application of these rodent models to future research may benefit the detection and treatment of ovarian cancer. PMID- 12911716 TI - Pseudomyxoma peritonei: the 'controversial' disease. AB - Pseudomyxoma peritonei (PMP) is a rare disease that is characterized by a large amount of mucinous ascites with peritoneal and omental implants. The etiology of the disease remains unclear. Histologically, two main categories have been described: disseminated peritoneal adenomucinosis (DPAM) and peritoneal mucinous carcinomatosis (PMCA). It is commonly diagnosed incidentally at laparotomy. Most investigators agree that radical surgical debulking and appendectomy are the cornerstone of treatment, but the optimal management of the disease remains controversial. The role of intraoperative and intraperitoneal chemotherapy has been evaluated by a number of authors. The clinical outcomes vary widely between the benign and the malignant forms and between the different treatment modalities. We discuss the pathology, origin, clinical presentation, diagnosis, treatment, and prognosis of PMP. PMID- 12911717 TI - Surgery and salvage chemotherapy for Chinese women with recurrent advanced epithelial ovarian carcinoma: a retrospective case-control study. AB - The objective of this paper is to clarify the role of cytoreductive surgery and salvage chemotherapy in the management of recurrent advanced epithelial ovarian carcinoma (RAEOC) and to identify factors affecting disease recurrence. One hundred sixty seven patients with RAEOC treated at the Cancer Hospital of Fudan University between January 1986 and December 1997 were retrospectively reviewed. Survival was calculated by Kaplan-Meier method with difference in survival estimated by the log-rank test. Independent prognostic factors were identified by the Cox stepwise regression model and variants associated with disease recurrence were determined using logistic stepwise regression methods. The median age was 52 (range 27-72) years. Sixty (35.9%) patients underwent re-debulking surgery, 23 of them with residual disease 43 degrees C). In dissociated dorsal root ganglion neurons (<30 micro m) from adult rat we demonstrated, using perforated patch clamp recording, that the inward current elicited in response to noxious heating is enhanced by nerve growth factor and reduced by capsazepine. The tachyphylaxis observed in response to the second of two heat pulses was reversed in most cells when nerve growth factor was introduced into the medium during the 5 min between the two heat stimuli, similar to findings using capsaicin [X. Shu & L.M. Mendell (1999) Neurosci. Lett.274, 159-162]. The threshold temperature did not change systematically after nerve growth factor. Using antibodies to TRPV1 and trkA in a subset of cells from which we recorded, we found a virtually perfect correlation between expression of TRPV1 and sensitivity to noxious heat. In addition, trkA expression was perfectly correlated with the ability of nerve growth factor to reverse tachyphylaxis. Thus, this physiological test is a reliable measure of trkA expression in cells sensitive to noxious heat. In agreement with studies in heterologous cells expressing trkA and TRPV1, pharmacologically blocking phospholipase C abolished the effect of nerve growth factor on heat-evoked currents in cells verified to express trkA. We conclude that the response of dorsal root ganglion neurons to noxious heat is conditioned by nerve growth factor in the same way as their response to capsaicin and that these responses require the presence of trkA and TRPV1. PMID- 12911750 TI - Heterogeneity and regulation of cellular prion protein glycoforms in neuronal cell lines. AB - The normal cellular prion protein is a small sialoglycoprotein highly expressed in neurons, the physiological function of which is largely unknown. Due to extensive N-glycosylations with a wide range of oligosaccharides, the prion protein displays a complex glycosylation pattern that could be of relevance for its function. The cellular prion protein patterns in adult mouse and rat brain, and in neuronal cell lines, appeared highly heterogeneous, as distinct levels and glycoforms of cellular prion protein were revealed by immunoblotting of corresponding samples. Amongst neuronal cell lines, mouse N2a neuroblastoma cells expressed low levels of endogenous prion protein. Mouse hypothalamic GT1-7 cells and rat pheochromocytoma PC-12 cells expressed highly glycosylated forms of cellular prion protein that were found neither in adult mouse and rat brain, nor in mouse brain during development. In contrast, rat B104 neuroblastoma cells abundantly expressed N-glycosylated cellular prion protein forms similar to those observed in mouse and rat brain. In all these cell lines, the prion protein was normally exported to and expressed at the outer cell membrane. Our results suggest that B104 cells may represent an appropriate cell model to investigate the physiological role of cellular prion protein in further detail as they highly express the normal 'brain-like' cellular prion protein glycoforms. In addition, we observed that the various prion glycoforms in B104 cells were tightly regulated both as a function of cell density and during neuronal differentiation, implying a potential role of cellular prion protein in cell-cell interactions and differentiation. PMID- 12911751 TI - Corticotropin-releasing factor receptor types 1 and 2 are differentially expressed in pre- and post-synaptic elements in the post-natal developing rat cerebellum. AB - Corticotropin-releasing factor (CRF)-like proteins act via two G-protein-coupled receptors (CRF-R1 and CRF-R2) playing important neuromodulatory roles in stress responses and synaptic plasticity. The cerebellar expression of corticotropin releasing factor-like ligands has been well documented, but their receptor localization has not. This is the first combination of a light microscopic and ultrastructural study to localize corticotropin-releasing factor receptors immunohistologically in the developing rat cerebellum. Both CRF-R1 and CRF-R2 were expressed in climbing fibres from early stages (post-natal day 3) to the adult, but CRF-R2 immunoreactivity was only prominent throughout the molecular layer in the posterior cerebellar lobules. CRF-R1 immunoreactivity was concentrated in apical regions of Purkinje cell somata and later in primary dendrites exhibiting a diffuse cytoplasmic appearance. In Purkinje cells, CRF-R1 immunoreactivity was never membrane bound post-synaptically in dendritic spines while CRF-R2 immunoreactivity was found on plasmic membranes of Purkinje cells from post-natal day 15 onwards. We conclude that the localization of these receptors in cerebellar afferents implies their pre-synaptic control of the release of corticotropin-releasing factor-like ligands, impacting on the sensory information being transmitted from afferents. Furthermore, the fact that CRF-R2 is membrane bound at synapses, while CRF-R1 is not, suggests that ligands couple to CRF-R2 via synaptic transmission and to CRF-R1 via volume transmission. Finally, the distinct expression profiles of receptors along structural domains of Purkinje cells suggest that the role for these receptors is to modulate afferent inputs. PMID- 12911752 TI - Involvement of platelet-activating factor and LIS1 in neuronal migration. AB - Platelet-activating factor (PAF, 1-O-alkyl-2-acetyl-sn-glycero-3-phosphocholine) is a biologically active lipid mediator. We have previously shown the expression of PAF receptor in neurons and microglia. PAF is produced in the brain from its precursor, and degraded by the enzyme PAF acetylhydrolase. LIS1 is a regulatory subunit of PAF acetylhydrolase, and is identical to a gene whose deletion causes the human neuronal migration disorder, type I lissencephaly. Indeed, Lis1 mutant mice display defects in neuronal migration and layering in vivo, and also in cerebellar granule cell migration in vitro. However, the roles of PAF and the PAF receptor in the neuronal migration remain to be determined. Here, we show that PAF receptor-deficient mice exhibited histological abnormalities in the embryonic cerebellum. PAF receptor-deficient cerebellar granule neurons migrated more slowly in vitro than wild-type neurons, consistent with the observation that a PAF receptor antagonist reduced the migration of wild-type neurons in vitro. Synergistic reduction of neuronal migration was observed in a double mutant of PAF receptor and LIS1. Unexpectedly, PAF affected the migration of PAF receptor deficient neurons, suggesting a receptor-independent pathway for PAF action. The PAF receptor-independent response to PAF was abolished in granule neurons derived from the double mutant mice. Thus, our results suggest that the migration of cerebellar granule cells is regulated by PAF through receptor-dependent and receptor-independent pathways, and that LIS1 is a pivotal molecule that links PAF action and neuronal cell migration both in vivo and in vitro. PMID- 12911753 TI - A novel method of generating neuronal cell lines from gene-knockout mice to study prion protein membrane orientation. AB - The technology of gene knockout and transgenic mice has allowed the study of the role of genes and their proteins in animal physiology and metabolism. However, these techniques have often been found to be limited in that some genetic manipulations of mice led either to a fatal phenotype or to compensations that mask the loss of function of the target protein. The experimentation on neurons from transgenic mice is particularly critical in the study of key proteins that may be involved in neurodegeneration. The cell fusion technique has been implemented as a novel way to generate cell lines from prion protein knockout mice. Fusion between neonatal mouse neurons and a neuroblastoma cell line have led to a Prnp degrees / degrees cell line that facilitates the study of the knockout phenotype. These cells are readily transfectable and allowed us to study the expression of prion protein mutants on a PrP-knockout background. Using this cell line we have examined the effect of PrP mutations reported to alter PrPc to a transmembrane form. Our results suggest that these mutations do not create transmembrane forms of the protein, but block normal transport of PrP to the cell membrane. PMID- 12911754 TI - The medullary dorsal reticular nucleus enhances the responsiveness of spinal nociceptive neurons to peripheral stimulation in the rat. AB - Single-unit spinal recordings combined with application of glutamate into the medullary dorsal reticular nucleus were used to assess the action of this nucleus upon deep dorsal horn neurons in rats. Injection of high glutamate concentrations (10 and 100 mm) induced a dramatic and long-lasting increase of the responses of wide-dynamic range neurons to electrical stimulation of the sciatic nerve in the noxious range, without affecting ongoing discharges. Post-stimulus time histograms revealed that this increase concerned the post-discharge, but not A- or C-fibre-mediated responses, which remained unchanged independently of the stimulation frequency applied. The onset of the glutamate-induced response enhancement occurred with a concentration-dependent time delay and developed slowly until its maximum. These data indicate that the medullary dorsal reticular nucleus exerts a facilitating action upon deep dorsal horn wide-dynamic range neurons by enhancing their capacity to respond to peripheral stimulation through prolongation of their discharge. This action is accompanied by the strengthening of wind-up of deep dorsal horn wide-dynamic range neurons, hence providing a plausible substrate for chronic pain states. These results are in agreement with previous behavioural studies suggesting a pronociceptive role for the dorsal reticular nucleus [Almeida et al. (1996) Brain Res. Bull., 39, 7-15; Almeida et al. (1999) Eur. J. Neurosci., 11, 110-122], and support the involvement of a reverberating circuit, previously described in morphological studies [Almeida et al. (1993) Neuroscience, 55, 1093-1106; Almeida et al. (2000) Eur. J. Pain, 4, 373-387], which probably operates only at a certain threshold of activation. PMID- 12911755 TI - Age-related irreversible progressive nigrostriatal dopaminergic neurotoxicity in the paraquat and maneb model of the Parkinson's disease phenotype. AB - While advancing age is the only unequivocally accepted risk factor for idiopathic Parkinson's disease, it has been postulated that exposure to environmental neurotoxicants combined with ageing could increase the risk for developing Parkinson's disease. The current study tested this hypothesis by exposing C57BL/6 mice that were 6 weeks, 5 months or 18 months old to the herbicide paraquat, the fungicide maneb or paraquat + maneb, a combination that produces a Parkinson's disease phenotype in young adult mice. Paraquat + maneb-induced reductions in locomotor activity and motor coordination were age dependent, with 18-month-old mice most affected and exhibiting failure to recover 24 h post-treatment. Three months post-treatment, reductions in locomotor activity and deficits in motor coordination were sustained in 5-month-old and further reduced in 18-month-old paraquat + maneb groups. Progressive reductions in dopamine metabolites and dopamine turnover were greatest in 18-month-old paraquat + maneb and paraquat groups 3 months post-treatment. Increased tyrosine hydroxylase enzyme activity compensated for striatal tyrosine hydroxylase protein and/or dopamine loss following treatment in 6-week-old and 5-month-old, but not 18-month-old paraquat and paraquat + maneb mice. Numbers of nigrostriatal dopaminergic neurons were reduced in all age groups following paraquat alone and paraquat + maneb exposure, but these losses, along with decreases in striatal tyrosine hydroxylase protein levels, were progressive in 18-month-old paraquat and paraquat + maneb groups between 2 weeks and 3 months post-exposure. Collectively, these data demonstrate enhanced sensitivity of the ageing nigrostriatal dopamine pathway to these pesticides, particularly paraquat + maneb, resulting in irreversible and progressive neurotoxicity. PMID- 12911756 TI - Contrasting expression of Kv4.3, an A-type K+ channel, in migrating Purkinje cells and other post-migratory cerebellar neurons. AB - Kv4.3, an A-type K+ channel, is the only channel molecule showing anterior posterior (A-P) compartmentalization in the granular layer of mammalian cerebellum known so far. Kv4.3 mRNA has been detected from the posterior but not anterior granular layer in adult rat cerebellum. To characterize this A-P compartmentalization further, we examined Kv4.3 protein expression in rat cerebellum by immunohistochemistry at the embryonic, early postnatal and adult stages. Specificity of the Kv4.3 antibody was confirmed by both Western blot and immunoprecipitation analysis. In adulthood, Kv4.3 was detected from the somatodendritic domain of posterior granule cells, with a restriction boundary in the vermal lobule VI extending laterally to the hemispheric crus 1 ansiform lobules. At the early postnatal stage, this A-P pattern first appeared on postnatal day 8, when significant numbers of granule cells had migrated into the posterior granular layer and started to express Kv4.3. Similar Kv4.3 expression in the somatodendritic domain of post-migratory neurons in the cerebellum was also observed in basket cells, stellate cells, a subset of GABAergic deep neurons, Lugaro cells and, probably, deep Lugaro cells. However, none of them showed A-P compartmentalization. Strikingly, we found Kv4.3 in several clusters of migrating Purkinje cells with mediolateral compartmentalization. These Purkinje cells no longer expressed Kv4.3 after completing the migration. By contrasting the expression in migrating and post-migratory neurons, our results suggest that Kv4.3 may play an important role in the development of cerebellum, as well as in the mature cerebellum. PMID- 12911757 TI - Altered responses to orexigenic (AGRP, MCH) and anorexigenic (alpha-MSH, CART) neuropeptides of paraventricular hypothalamic neurons in early postnatally overfed rats. AB - Food intake and energy expenditure are regulated by neuropeptides in the hypothalamus. While cocaine- and amphetamine-regulated transcript (CART) peptide and melanocortins such as alpha-melanocyte-stimulating hormone (alpha-MSH) are anorexigenic and increase energy expenditure, the endogenous melanocortin receptor antagonist agouti gene-related protein (AGRP), melanin-concentrating hormone (MCH) and neuropeptide Y (NPY) are orexigenic, anabolic peptides. Alterations in the regulatory balance may promote excessive weight gain. The action of these peptides on paraventricular hypothalamic neurons was studied in brain slices of overweight, adult rats previously subjected to early postnatal overfeeding in small litters of only three pups per mother, compared to 12 pups per dam in control litters. CART, melanocortins and NPY significantly excited paraventricular neurons of controls, whereas neurons of small-litter rats were mainly inhibited. Inhibition was dominant following administration of AGRP, MCH and NPY. The altered responses of paraventricular neurons in adult small-litter rats might reflect a general mechanism of neurochemical plasticity and 'malprogramming' of hypothalamic neuropeptidergic systems acquired during the postnatal critical differentiation period, thus leading to permanently altered function of these regulatory systems of body weight. PMID- 12911758 TI - Low molecular weight constituents of male mouse urine mediate the pregnancy block effect and convey information about the identity of the mating male. AB - Mouse urine contains a complex mixture of chemosignals including a variety of small volatile molecules that are bound to major urinary proteins. In addition to signalling maleness, male urine also conveys information about individuality, which allows recently mated female mice to distinguish the urinary chemosignals of the mating male from those of an unfamiliar male. The highly polymorphic nature of the major urinary proteins makes them a likely candidate for conveying individuality information in the context of the pregnancy block effect. This was investigated by comparing the pregnancy-blocking effectiveness of a high molecular weight urinary fraction, containing major urinary proteins, with that of a low molecular weight fraction containing volatile ligands. Not only was the high molecular weight fraction ineffective in blocking pregnancy, but it also appeared to be less important in signalling individuality than the low molecular fraction. The high molecular weight fraction was ineffective in inducing expression of the immediate early gene product egr-1 in the accessory olfactory bulb. In contrast, the low molecular weight fraction induced egr-1 expression in the mitral/tufted neurons in the anterior subregion of the accessory olfactory bulb, suggesting that they activate the V1R class of vomeronasal receptor neuron. PMID- 12911759 TI - Connexin30-deficient mice show increased emotionality and decreased rearing activity in the open-field along with neurochemical changes. AB - Gap-junction channels in the brain, formed by connexin (Cx) proteins with a distinct regional/cell-type distribution, allow intercellular electrical and metabolic communication. In astrocytes, mainly the connexins 43, 26 and 30 are expressed. In addition, connexin30 is expressed in ependymal and leptomeningeal cells, as well as in skin and cochlea. The functional implications of the astrocytic gap-junctional network are not well understood and evidence regarding their behavioural relevance is lacking. Thus, we have tested groups of Cx30-/-, Cx30+/-, and Cx30+/+ mice in the open-field, an object exploration task, in the graded anxiety test and on the rotarod. The Cx30-/- mice showed reduced exploratory activity in terms of rearings but not locomotion in the open-field and object exploration task. Furthermore, Cx30-/- mice exhibited anxiogenic behaviour as shown by higher open-field centre avoidance and corner preference. Graded anxiety test and rotarod performance was similar across groups. The Cx30-/ mice had elevated choline levels in the ventral striatum, possibly related to their aberrant behavioural phenotypes. The Cx30+/- mice had lower dopamine and metabolite levels in the amygdala and ventral striatum and lower hippocampal 5 hydroxyindole acid (5-HIAA) concentrations relative to Cx30+/+ mice. Furthermore, the Cx30+/- mice had lower acetylcholine concentrations in the ventral striatum and higher choline levels in the neostriatum, relative to Cx30+/+ mice. Our data suggest that the elimination of connexin30 can alter the reactivity to novel environments, pointing to the importance of gap-junctional signalling in behavioural processes. PMID- 12911760 TI - Potentiation of amygdaloid and hippocampal auditory-evoked potentials in a discriminatory fear-conditioning task in mice as a function of tone pattern and context. AB - According to the local memory storage hypothesis, information about the tone shock association in an auditory fear-conditioning paradigm is stored in synapses within the lateral amygdala. Thus, fear-conditioning-induced potentiation of auditory-evoked potentials in response to a conditioned stimulus (CS+, a series of short lasting tones; patterned tone) has been interpreted as an in vivo correlate of amygdaloid synaptic plasticity. Here, we re-examine the specificity of potentiation of auditory-evoked potentials in terms of (i) local confinement to the lateral amygdala, (ii) parameters of CS+ and (iii) influence of context, using a discriminatory fear-conditioning paradigm. Adult male C57BL/6J mice were implanted with recording electrodes aimed at the lateral amygdala, the CA1 region of the hippocampus and the neck muscles for simultaneous recordings of auditory evoked potentials and startle responses. In a neutral context, auditory-evoked potentials within lateral amygdala and CA1 as well as startle and freezing responses to the CS+ were significantly potentiated following conditioning, as compared with pre-conditioning values and responses to a neutral stimulus (CSn; tone of different frequency). Potentiation was only evident if CS+ was presented as a uniform series but not if presented mixed with CSn. Accordingly, mice failed to show intensified freezing to a patterned tone if a single lasting tone of the same frequency served as CS+. Both CA1 and lateral amygdala auditory-evoked potentials were potentiated in response to CSn if presented in the conditioning context. These findings demonstrate that (i) potentiation of auditory-evoked potentials is not restricted to the lateral amygdala, (ii) both tone frequency and pattern of tone presentation are essential for proper CS+ recognition and (iii) contextual memory leads to a general potentiation of auditory-evoked potentials. PMID- 12911761 TI - A double dissociation between serial reaction time and radial maze performance in rats subjected to 192 IgG-saporin lesions of the nucleus basalis and/or the septal region. AB - The cholinergic basal forebrain has been implicated in aspects of cognitive function including memory and attention, but the precise contribution of its major components, the basalocortical and the septohippocampal systems, remains unclear. Rats were subjected to lesions of either the nucleus basalis magnocellularis (Basalis), the medial septum/vertical limb of the diagonal band of Broca (Septum), or both nuclei (Basalis + Septum), using the selective cholinotoxin 192 IgG-saporin. Cognitive performance was evaluated in tasks taxing attention (the five-choice serial reaction time task, 5-CSRTT) and spatial working memory (radial arm maze, RAM). Nucleus basalis lesions disrupted performance of the 5-CSRTT, as demonstrated by decreased choice accuracy, increased incidence of missed trials, increased latencies to respond correctly, and a disrupted pattern of response control. Combined lesions of the Basalis and Septum resulted in qualitatively similar deficits to Basalis lesions alone, although interestingly, these rats were unimpaired on measures of response speed, and showed weaker deficits on accuracy and omissions. Decreasing the attentional load by lengthening stimulus duration reversed some of the deficits in Basalis and Basalis + Septum rats, suggesting an attentional deficit rather than motivation or motor perturbations. Performance in rats with septal lesions was only affected when task difficulty was increased. In the RAM an opposing pattern of effects was observed, with Septum and Basalis + Septum rats showing dramatic impairments, and Basalis rats performing normally. Taken together, these data provide clear evidence for a functional dissociation between septohippocampal and basalocortical cholinergic systems in aspects of cognitive function. PMID- 12911762 TI - A role for the periaqueductal grey in opioidergic inhibition of maternal behaviour. AB - Opiates are known to be involved in the regulation of various events surrounding parturition and lactation, such as maternal behaviour in rats. The onset of this behaviour has been closely linked to opiate action in the medial pre-optic area, where administration of morphine disrupts maternal behaviour during lactation. By combining the use of Fos protein immunohistochemical detection and pharmacological manipulations, in the present paper we show that the periaqueductal grey (PAG) is another region critically involved in the opioidergic blockade of maternal behaviour. According to our observations, a critical level of morphine-induced activation of the rostral lateral PAG appears to be required to inhibit maternal behaviour in lactating rats. This hypothesis was further confirmed in experiments showing that morphine's inhibitory effect on maternal responsiveness was blocked by unilateral naloxone injection into the rostral PAG, but not into nearby regions of the mesencephalic reticular nucleus. Therefore, only a partial inhibition of the opiate's effect on the rostral PAG was needed to block the inhibitory effect of morphine on maternal behaviour. Further studies are needed to ascertain whether the rostral lateral PAG plays a role in the natural onset of maternal behaviour, playing a complementary role to the medial pre-optic area, or merely inhibits maternal behaviour in response to this specific pharmacological challenge. Conversely, the present findings may well reflect a more general role of the PAG, seemingly providing an important piece of information for proposing a hitherto unexplored concept of the PAG as an important centre for the selection of adaptive behavioural responses. PMID- 12911763 TI - Changes of effective connectivity between the lateral and medial parts of the prefrontal cortex during a visual task. AB - Structural equation modelling was used to study the change of connectivity during a visual task with continuous variation of the attention load. The model was based on areas defined by the haemodynamic responses described elsewhere [Mazoyer, P., Wicker, B. & Fonlupt, P. (2002) A neural network elicited by parametric manipulation of the attention load. Neuroreport, 13, 2331-2334], including occipitotemporal, parietal, temporal and prefrontal (lateral and medial areas) cortices. We have studied stationary- (which does not depend on the attentional load) and attention-related coupling between areas. This allowed the segregation of two subsystems. The first could reflect a system performing the integration step of the visual signal and the second a system participating in response selection. The major finding is the mutual negative influence between the lateral and medial parts of the prefrontal cortex. This negative influence between these two brain regions increased with the attention load. This is interpreted as a modification of the balance between integration and decision processes that are needed for the task to be efficiently completed. PMID- 12911764 TI - Neural activity related to the processing of increasing monetary reward in smokers and nonsmokers. AB - This study investigated the processing of increasing monetary reward in nonsmoking and smoking subjects. The choice of the subject populations has been motivated by the observation of differences between nonsmokers and smokers in response to rewarding stimuli in a previous study. Subjects performed a pattern recognition task with delayed response, while rCBF was measured with [H215O] PET. Correct responses to the task were reinforced with three different amounts of monetary reward. The subjects received the sum of the rewards at the end of the experiment. The results show that a cortico-subcortical loop, including the dorsolateral prefrontal cortex, the orbitofrontal cortex, the cingulate gyrus and the thalamus is involved in processing increasing monetary reward. Furthermore, the striatal response differentiates nonsmokers from smokers. Thus, we found significant correlations between rCBF increases in striatum and increasing monetary reward and between striatal rCBF increases and mood in nonsmokers, but not in smokers. Moreover, no significant mood changes among the different monetary rewards could be observed in smokers. We infer that the response of the striatum to reward is related to changes in subjective feelings. The differences between smokers and nonsmokers confirm our previous conclusions that the association between blood flow, performance, mood and amount of reward is more direct in nonsmokers. PMID- 12911765 TI - Normal prism adaptation but reduced after-effect in basal ganglia disorders using a throwing task. AB - Prism adaptation is a form of visuomotor learning in which the visual and motor systems need to be adjusted because a visual perturbation is produced by horizontally displacing prisms. Despite being known for over two centuries, the neuronal substrates of this phenomenon are not yet completely understood. In this article the possible role of the basal ganglia in this kind of learning was analysed through a study of Huntington's and Parkinson's disease patients. A throwing technique requiring the use of open loop feedback was used. The variables analysed were visuomotor performance, adaptation rate and magnitude, and the after-effect. The results clearly showed that both Huntington's and Parkinson's disease groups learned at the same rate as control subjects. In addition, despite having a disturbed visuomotor performance, both experimental groups showed the same adaptation magnitude as the control group. Finally, the after-effect, which is measured after removing the prisms, is reduced in both patients groups. This reduction leads to a disruption in the normal adaptation after-effect correlation found in normal volunteers. These results suggest that basal ganglia are not involved in this type of open-looped visuomotor learning. The large number of patients studied as well as the similarity of the findings between both populations support this hypothesis. By contrast, there is an impairment in the after-effect on both basal ganglia patient populations. This impairment may be the result of the deterioration of the perceptual recalibration process involved in visuomotor learning. PMID- 12911766 TI - Different representations of pleasant and unpleasant odours in the human brain. AB - Odours are important in emotional processing; yet relatively little is known about the representations of the affective qualities of odours in the human brain. We found that three pleasant and three unpleasant odours activated dissociable parts of the human brain. Pleasant but not unpleasant odours were found to activate a medial region of the rostral orbitofrontal cortex. Further, there was a correlation between the subjective pleasantness ratings of the six odours given during the investigation with activation of a medial region of the rostral orbitofrontal cortex. In contrast, a correlation between the subjective unpleasantness ratings of the six odours was found in regions of the left and more lateral orbitofrontal cortex. Moreover, a double dissociation was found with the intensity ratings of the odours, which were not correlated with the BOLD signal in the orbitofrontal cortex, but were correlated with the signal in medial olfactory cortical areas including the pyriform and anterior entorhinal cortex. Activation was also found in the anterior cingulate cortex, with a middle part of the anterior cingulate activated by both pleasant and unpleasant odours, and a more anterior part of the anterior cingulate cortex showing a correlation with the subjective pleasantness ratings of the odours. Thus the results suggest that there is a hedonic map of the sense of smell in brain regions such as the orbitofrontal cortex, and these results have implications for understanding the psychiatric and related problems that follow damage to these brain areas. PMID- 12911767 TI - Language perception activates the hand motor cortex: implications for motor theories of speech perception. AB - The precise mechanisms of how speech may have developed are still unknown to a large extent. Gestures have proven a powerful concept for explaining how planning and analysing of motor acts could have evolved into verbal communication. According to this concept, development of an action-perception network allowed for coding and decoding of communicative gestures. These were manual or manual/articulatory in the beginning and then became increasingly elaborate in the articulatory mode. The theory predicts that listening to the 'gestures' that compose spoken language should activate an extended articulatory and manual action-perception network. To examine this hypothesis, we assessed the effects of language on cortical excitability of the hand muscle representation by transcranial magnetic stimulation. We found the hand motor system to be activated by linguistic tasks, most notably pure linguistic perception, but not by auditory or visuospatial processing. The amount of motor system activation was comparable in both hemispheres. Our data support the theory that language may have evolved within a general and bilateral action-perception network. PMID- 12911768 TI - Direction-selective motion blindness after unilateral posterior brain damage. AB - Motion blindness (MB) is defined as the selective disturbance of visual motion perception despite intact perception of other features of the visual scene. MB is characterized by a pandirectional deficit of motion direction discrimination and is assumed to result from damage to the visual motion pathway, especially area MT/V5. However, the most characteristic feature of primate MT/V5 neurons is not their motion selectivity but their preference for one direction of motion (direction selectivity), which changes incrementally at neighbouring columns. In addition to this microscopic directional organization, studies in nonhuman and human primates suggest that single directions of motion are also coded at a more macroscopic level. We thus hypothesized that if MB in humans results from damage to direction-selective neurons in the visual motion pathway, posterior brain damage might cause MB which is direction selective, not pandirectional. The present study investigated motion direction discrimination in patients with posterior unilateral brain damage and determined separate psychophysical thresholds for the four cardinal directions. In addition, we analysed whether the direction of erroneous motion perception (i.e. the perception of right motion for upward motion) was random or showed a directional bias. We report three principal findings. First, motion direction discrimination was severely impaired in one or two directions while it was normal in the other directions. This constituted direction-selective MB. Second, MB was characterized not only by a quantitative direction-selective increase in psychophysical thresholds but also by a qualitative impairment of perceiving motion direction systematically in wrong directions. Both findings suggest that the cortical modules specialized for the perception of a single direction of motion might be larger than previously thought. Third, lesion analysis showed that unilateral damage, not only the human homologue of MT/V5 but also to parieto-occipital cortex, leads to MB. PMID- 12911769 TI - Alpha 2 adrenergic receptors on GABAergic, putative sleep-promoting basal forebrain neurons. AB - The basal forebrain plays an important role in the modulation of cortical activity and sleep-wake states. Yet its role must be multivalent as lesions reportedly diminish cortical fast activity and also cortical slow activity along with slow wave sleep (SWS). Basal forebrain cholinergic vs. GABAergic cell groups could differentially influence these processes. By labelling recorded neurons with Neurobiotin (Nb) using the juxtacellular technique and identifying them by immunostaining, we previously found that whereas all cholinergic cells increased their firing, the majority of GABAergic neurons decreased their firing in association with evoked cortical activation in urethane-anaesthetized rats. Here, we examined the possibility that such GABAergic, cortical activation 'off' cells might bear alpha 2 adrenergic receptors (alpha2AR) through which noradrenaline (NA) could inhibit them during cortical activation. First using simple dual immunostaining for glutamic acid decarboxylase (GAD) and the alpha2AAR, we found that the majority (approximately 60%) of GAD-immunopositive (GAD+) neurons through the magnocellular preoptic nucleus (MCPO) and substantia innominata (SI) were labelled for the alpha2AAR. Second, in urethane-anaesthetized rats, we examined whether Nb-labelled, GAD+ cortical activation 'off' neurons that discharged maximally in association with cortical slow wave activity, were immunopositive for alpha2AAR. We found that all the Nb+/GAD+'off' cells were labelled for the alpha2AAR. Such cells could be inhibited in association with cortical activation and waking when noradrenergic locus coeruleus (LC) neurons discharge and be disinhibited with cortical slow waves and SWS when these neurons become inactive. We thus propose that alpha2AR-bearing GABAergic basal forebrain neurons constitute sleep-active and sleep-promoting neurons. PMID- 12911770 TI - Neural substrates of microstimulation-evoked tingling: a chronaxie study in human somatosensory thalamus. AB - Intra-operative micro-electrode stimulation of sensorimotor thalamus produces paraesthesia or tingling in various body regions and is used to map somatotopy prior to implantation of deep brain-stimulating electrodes in awake patients. The neural elements affected by such microstimulation are unknown. Using paraesthesia as the behavioural-physiological response threshold, we measured chronaxie times for microstimuli applied to both somatosensory thalamic nuclei (cellular region) and its axonal afferents, the medial lemniscus. White matter chronaxie times were relatively unimodal, whereas two different clusters of chronaxie times were identified in grey matter: one corresponding to that of the medial lemniscus and the other about five times longer and compatible with that obtained from cell somata. Therefore, excitations of local axons and/or cell bodies can both contribute to the paraesthesia evoked during intra-operative thalamic mapping. PMID- 12911771 TI - Managing anaphylaxis: effective emergency and long-term care are necessary. PMID- 12911772 TI - Optimizing the diagnosis of peanut and tree nut allergy. PMID- 12911773 TI - Interleukin-18 in asthma and other allergies. PMID- 12911774 TI - Oncostatin M: an interleukin-6-like cytokine relevant to airway remodelling and the pathogenesis of asthma. PMID- 12911775 TI - Anaphylaxis: risk factors for recurrence. AB - BACKGROUND: There are few studies on the incidence or recurrence of anaphylaxis. OBJECTIVE: To examine the incidence of anaphylaxis and risk factors for recurrence. METHODS: A prospective study of 432 patients referred to a community based specialist practice in the Australian Capital Territory with anaphylaxis, followed by a survey to obtain information on recurrence. RESULTS: Of 432 patients (48% male, 73% atopic, mean 27.4 years, SD 19.5, median 26) with anaphylaxis, 260 patients were seen after their first episode; 172 experienced 584 previous reactions. fifty-four percent of index episodes were treated in hospital. Aetiology was identified in 91.6% patients: food (61%), stinging insects (20.4%) or medication (8.3%). The minimum occurrence and incidence of new cases of anaphylaxis was estimated at 12.6 and 9.9 episodes/100,000 patient years, respectively. Follow-up data were obtained from 304 patients (674 patient years). One hundred and thirty experienced further symptoms (45 serious), 35 required hospitalization and 19 administered adrenaline. Accidental ingestion of peanut/tree nut caused the largest number of relapses, but the highest risk of recurrence was associated with sensitivity to wheat and/or exercise. Rates of overall and serious recurrence were 57 and 10 episodes/100 patient-years, respectively. Of those prescribed adrenaline, 3/4 carried it, 2/3 were in date, and only 1/2 patients faced with serious symptoms administered adrenaline. Five patients each developed new triggers for anaphylaxis, or re-presented with significant psychiatric symptoms. CONCLUSION: In any 1 year, 1/12 patients who have suffered anaphylaxis will experience recurrence, and 1/50 will require hospital treatment or use adrenaline. Compliance with carrying and using adrenaline is poor. Occasional patients develop new triggers or suffer psychiatric morbidity. PMID- 12911776 TI - Interpretation of tests for nut allergy in one thousand patients, in relation to allergy or tolerance. AB - BACKGROUND: Peanut and tree nut allergy are common, increasing in prevalence and the commonest food cause of anaphylaxis. In the USA, 7.8% are sensitized (have nut-specific IgE), but not all those sensitized are allergic. Lack of data makes interpretation of tests for nut-specific IgE difficult. OBJECTIVES: This is the first study to investigate the clinical significance of test results for peanut and tree nut allergy in allergic or tolerant patients. Findings are related to the severity of the allergy. METHOD: An observational study of 1000 children and adults allergic to at least one nut. History of reactions (severity graded) or tolerance to up to five nuts was obtained and skin prick test (SPT)/serum specific IgE (CAP) performed. RESULTS: There was no correlation between SPT size and graded severity of worst reaction for all nuts combined or for peanut, hazelnut, almond and walnut. For CAP, there was no correlation for all nuts. Where patients tolerated a nut, 43% had positive SPT of 3-7 mm and 3% > or = 8 mm. For CAP, 35% were positive (0.35-14.99 kU/L) and 5% > or = 15 kU/L. In SPT range 3-7 mm, 54% were allergic and 46% were tolerant. There was poor concordance between SPT and CAP (66%). Of patients with a clear nut-allergic history, only 0.5% had negative SPT, but 22% negative CAP. CONCLUSIONS: Magnitude of SPT or CAP does not predict clinical severity, with no difference between minor urticaria and anaphylaxis. SPT is more reliable than CAP in confirming allergy. Forty-six per cent of those tolerant to a nut have positive tests > or = 3 mm (sensitized but not allergic). One cannot predict clinical reactivity from results in a wide 'grey area' of SPT 3-7 mm; 22% of negative CAPs are falsely reassuring and 40% of positive CAPs are misleading. This emphasizes the importance of the history. Understanding this is essential for accurate diagnosis. Patients with SPT > or = 8 mm and CAP > or = 15 kU/L were rarely tolerant so these levels are almost always (in > or = 95%) diagnostic. PMID- 12911777 TI - Thresholds of clinical reactivity to milk, egg, peanut and sesame in immunoglobulin E-dependent allergies: evaluation by double-blind or single-blind placebo-controlled oral challenges. AB - BACKGROUND: The prevalence of food anaphylaxis due to masked allergens has increased within the last 10 years. Contamination of manufactured products by food allergens is a key concern for food industries. OBJECTIVE: To determine quantities eliciting reactions in patients who have an IgE-dependent food allergy, thanks to standardized oral provocation tests. To evaluate the subsequent levels of sensitivity required for the detection tests of allergens for egg, peanut, milk and sesame. METHODS: Prick-in-prick tests, Cap system RAST, and single or double-blind placebo-controlled food challenges (SBPCFC or DBPCFC) were performed. The doses of natural food were gradually increased from 5 to 5000 mg for solid food and from 1 to 30 mL for peanut oil, sunflower oil, soy oil and sesame oil. RESULTS: Data from 125 positive oral challenges to egg, 103 to peanut, 59 to milk and 12 to sesame seeds were analysed. Haemodynamic modifications were observed in 2%, 3%, 1.7%, and 8% of the oral challenges (OCs) to egg, peanut, milk and sesame, respectively. Respiratory symptoms were observed in 12%, 20%, 10% and 42% of egg, peanut milk and sesame allergies, respectively. A cumulative reactive dose inferior or equal to 65 mg of solid food or 0.8 mL of milk characterized 16%, 18%, 5% and 8% of egg, peanut, milk and sesame allergies, respectively. 0.8% of egg allergies, 3.9% of peanut allergies, and 1.7% of milk allergies reacted to 10 mg or less of solid food or to 0.1 mL for milk. The lowest reactive threshold has been observed at less than 2 mg of egg; 5 mg of peanut, 0.1 mL of milk and 30 mg of sesame seed. Ten out of 29 OC with peanut oil, two out of two OC with soy oil and three out of six OC with sunflower oil were positive. Five out six OC with sesame oil were positive: 1 and 5 mL induced an anaphylactic shock. CONCLUSION: The risk of asthma and anaphylactic shock to sesame and peanut is confirmed. Minimal reactive quantities show that, in order to guarantee a 95% safety for patients who are allergic to egg, peanut and milk, and on the basis of consumption of 100 g of food, the detection tests should ensure a sensitivity of 10 p.p.m. for egg, 24 p.p.m. for peanut and 30 p.p.m. for milk proteins. Oil allergies being considered, the limit of sensitivity should fall to 5 p.p.m. PMID- 12911778 TI - A kinetic study in adults with food hypersensitivity assessed as eosinophil activation in fecal samples. AB - BACKGROUND: Immune-mediated food hypersensitivity affecting the gut is difficult to evaluate, and objective tools to diagnose local gastrointestinal (GI) inflammatory reactions are lacking. OBJECTIVES: To determine whether allergic manifestations in adults with a history of food-related GI symptoms could be assessed in feces during symptomatic and non-symptomatic periods, using the surrogate markers, eosinophil cationic protein (ECP), eosinophil protein X (EPX) and myeloperoxidase (MPO). METHODS: Thirteen subjects with food hypersensitivity related GI symptoms, confirmed by a positive double-blind placebo-controlled food challenge (DBPCFC), were subjected to an open kinetic food challenge design for 6 weeks. Symptoms were recorded and scored during the 3-week study period and stool samples were obtained every day. The surrogate markers ECP, EPX and MPO were measured in the supernatants from feces samples. RESULTS: A significant increase in abdominal pain, distension and flatulence was observed during challenge, with a gradual decrease during elimination diet. Both between days and subjects, EPX levels were more frequently increased compared to ECP and MPO. Individuals with a history of a short duration of symptoms had significantly higher mean levels of EPX and MPO than those with a longer duration of symptoms. CONCLUSIONS: An overall increase in levels of eosinophil markers, in particular EPX, was observed in feces from patients with food-related GI symptoms. However, rather than being a tool to differentiate symptomatic from non-symptomatic periods, EPX might be used for detecting an ongoing clinical or subclinical chronic inflammation, that may have an impact on the patient's clinical course of GI symptoms. PMID- 12911779 TI - Concurrent cereal allergy in children with cow's milk allergy manifested with atopic dermatitis. AB - BACKGROUND: There is increasing consensus about the significance of food allergens in the pathogenesis of atopic dermatitis (AD) in infancy and childhood, with cow's milk and egg accounting for most of the reactions. Previous studies have indicated that multiple food sensitization, such as cereals, is very common in patients with cow's milk allergy (CMA). Evidence is lacking, however, as to its clinical relevance. OBJECTIVE: The purpose of this study was to determine the concurrent occurrence of cereal allergy among children with challenge-proven CMA who have residual symptoms, such as AD and/or gastrointestinal symptoms, during cow's milk elimination diet. Further, we sought to evaluate the utility of patch testing in prescreening foods other than cow's milk behind allergic symptoms in children. METHODS: The study population comprised 90 children, aged from 2.5 to 36 months (mean 1.1 years), with challenge-proven CMA. As a result of residual symptoms during meticulous cow's milk elimination diet (AD: n=80, and gastrointestinal: n=10), the children were put on a cereal elimination diet (oats, wheat, rye, and barley) and skin prick tests (SPT) and patch testing with cereals were performed. Open cereal challenge was performed to confirm cereal allergy. RESULTS: Cereal challenge was positive in 66 (73%) of the children with CMA. Of them, 17% reacted with immediate reactions and delayed-onset reactions were seen in 83% of the children. SPT was positive in 23%, patch test in 67%, and either SPT or patch test was positive in 73% of the children with cereal allergy. SPT gave the best positive predictive value, whereas SPT together with patch test gave the best negative predictive value. CONCLUSIONS: Residual symptoms, such as eczema or gastrointestinal symptoms in CMA children may be a sign of undetected allergy to other food antigens. SPT with cereals aids in diagnosing cereal allergy in small children, especially when used together with patch testing. PMID- 12911780 TI - Epicutaneous exposure to protein antigen and food allergy. AB - BACKGROUND: The aetiology of food allergy remains unclear. Although failure to develop or breakdown in oral tolerance has been proposed, the existence of physiologic sensitization routes other than the gastrointestinal tract cannot be excluded. OBJECTIVE: The purpose of this study is to clarify whether or not exposure to allergen through the skin can promote food allergy. METHODS: BALB/c mice were shaved on the back, and a patch impregnated with 100 micro g of ovalbumin (OVA) was applied to the dorsal skin for a 1-week period and then removed. After three courses of sensitization, OVA-specific antibodies in sera were measured, and then mice were orally challenged with 50 mg of OVA. Anaphylactic symptoms, plasma histamine levels, and histology of intestines and lungs after oral challenge were examined. RESULTS: Epicutaneous (EC) sensitization of mice to OVA induced a high level of OVA-specific IgE. Subsequent oral challenge with OVA resulted in symptoms of systemic anaphylaxis with elevated levels of plasma histamine as well as histological changes in both intestines and lungs. In the presence of anti-IL-4 antibodies, EC sensitization failed to provoke an IgE response, but still induced a Th2-predominant cellular immune response in lungs after oral challenge. CONCLUSION: We demonstrated for the first time that food allergy can be induced by allergen exposure through the skin. Our results identify a novel role of EC sensitization in the pathogenesis of food allergy. PMID- 12911781 TI - Usefulness of specific immunotherapy in patients with severe perennial allergic rhinitis induced by house dust mite: a double-blind, randomized, placebo controlled trial. AB - OBJECTIVES: To evaluate the effectiveness of specific immunotherapy (SIT) in patients with severe house dust mite (HDM)-induced perennial allergic rhinitis using diary cards and objective endpoints. PATIENTS AND METHODS: Thirty-six adult patients were selected with moderate to severe allergic rhinitis due to HDM allergy uncontrolled by regular anti-allergic drugs. Twenty-eight patients completed the study, 22 of these patients also had mild asthma. Subjects were stratified for HDM sensitivity on the basis of their 4-week diary card score and the size of their immediate and late-phase skin reaction to HDM. The groups were well matched for all relevant parameters. Patients were randomized to receive active preparation (Alutard(R)-SQ, ALK, Dermatophagoides pteronyssinus extract) or an identical placebo preparation. Increasing doses were administered until the maintenance dose was reached. This dose was then given once a month for 12 months. RESULTS: Clinical efficacy was evaluated by symptom medication diary cards recorded for 4 weeks after 12 months of continuous treatment and compared with pre-treatment scores. Skin test reactivity was re-measured after 12 months of treatment to HDM, cat dander and codeine phosphate. After 1 year of treatment, the actively treated group showed a 58% reduction in diary card symptom scores (P<0.002) and a 20% reduction in the use of rescue medication. The placebo group had a 32% reduction in symptom scores (P=NS), but no reduction in rescue medication requirements. The active group showed 36% reduction in skin prick test sensitivity to D. pteronyssinus (P=0.006), while the placebo group values were unchanged. Skin reactivity to codeine was unchanged in both groups. No significant adverse reactions to SIT were encountered. CONCLUSIONS: One year of SIT for D. pteronyssinus in patients with poorly controlled rhinitis (+/-mild asthma) produced clinically useful improvement as shown by symptom-medication diary cards and reductions in immediate skin reactions compared with placebo treatment. PMID- 12911782 TI - Mycobacterium vaccae administration during allergen sensitization or challenge suppresses asthmatic features. AB - BACKGROUND AND OBJECTIVE: The hygiene hypothesis suggests that a lack of bacterial infections would favour the development of allergic disease. For this reason, bacteria or their components can be used as potential treatment for allergic asthma. We investigated whether heat-killed Mycobacterium vaccae is either able to suppress the induction of allergic asthma or able to suppress already established allergic asthma. METHODS: Mice were sensitized with ovalbumin (OVA)/alum on days 0 and 14. Thereafter, mice were challenged on days 35, 39 and 42 by inhalation of either OVA or saline aerosols. M. vaccae-treated mice received an injection with 106, 107 or 108 CFU heat-killed M. vaccae on days 0 and 14 or 107 CFU on days 35 and 39. On day 43, the airway responsiveness of the mice to increasing concentrations of methacholine was assessed, blood was withdrawn to measure serum parameters, and lung lavage was performed to detect cytokines and inflammatory cell number. RESULTS: Treatment of OVA-sensitized mice with 107 CFU M. vaccae either during sensitization or challenge suppresses airway hyper-responsiveness, airway eosinophilia and IL-5 production after OVA challenge. The increases in OVA-specific serum IgE and in IL-4 by respiratory challenges with OVA were only diminished after M. vaccae treatment (107 CFU) during sensitization. CONCLUSIONS: Heat-killed M. vaccae prevents allergic and asthmatic manifestations in a mouse model and, more importantly, M. vaccae treatment during challenge suppresses features of asthma, which opens up possibilities for new therapeutic interventions. PMID- 12911783 TI - Methotrexate therapy in asthma increases T cell susceptibility to corticosteroid inhibition. AB - BACKGROUND: Concomitant methotrexate (MTX) therapy of severe, oral corticosteroid dependent asthmatics has been shown to be corticosteroid sparing, but the mechanism is unknown. We hypothesized that MTX therapy of these patients increases the susceptibility of their T cells to corticosteroid inhibition. OBJECTIVE: To measure prednisolone inhibition of lectin-induced proliferation of peripheral blood T cells from a group of these patients before, during and following MTX therapy. METHODS: Eighteen severe asthmatics (median (range) age 56 (33-68) years, FEV1 61 (38-69)% predicted, dependent on oral prednisolone 15 (7.5 25) mg/day in addition to high-dose, inhaled corticosteroids) were treated with MTX 15 mg intramuscularly, weekly for 28 weeks. After 12 weeks of MTX, oral prednisolone dosages were reduced systematically over 16 weeks, provided that asthma control did not deteriorate. Patients were followed for a further 12 weeks after MTX withdrawal. Concentration-dependent, prednisolone inhibition of lectin induced proliferation of peripheral blood T cells was measured just prior to MTX therapy (week 1) and at weeks 12, 28 and 40, and IC50 concentrations were interpolated. RESULTS: By week 28 of MTX therapy, patients were able to reduce oral prednisolone dosages from (median, SIQR) 15 (10-20.5) to 5.9 (1.4-9.4) mg/day (P<0.01) without alteration of lung function and symptoms, while median IC50 values for prednisolone inhibition of peripheral blood T cell proliferation were reduced from 49 (21-144) to 4 (1-9) nm (P<0.0001). These increased again to 15 (9.4-25.7) mg/day and 36 (18-67) nm, respectively, following MTX withdrawal. A correlation (P<0.01) was observed between percentage reductions in prednisolone dosages in vivo and fold changes in prednisolone IC50in vitro between weeks 12 and 28. CONCLUSION: This effect of MTX may at least partly account for its oral corticosteroid-sparing effect in severe asthma. PMID- 12911784 TI - Association between interleukin-18 gene polymorphism 105A/C and asthma. AB - BACKGROUND: IL-18 has been shown to exert anti-allergic or allergy-promoting activities, but the existence of genetic polymorphisms in the coding regions of IL-18 gene has not been demonstrated. OBJECTIVE: The aim of this study was to investigate whether polymorphism is present in the coding regions of the IL-18 gene and, if so, to further analyse the association between polymorphism and asthma in a case-control study. METHODS: We screened the coding regions of the IL 18 gene for polymorphisms by using PCRsingle-stranded conformation polymorphism and direct sequencing of PCR products, followed by analysis of the association between polymorphism and asthma. RESULTS: We identified one polymorphism (105A/C) in the coding regions. The frequency of the 105A allele was significantly higher in asthmatic patients than in controls (P<0.01; odds ratio (OR)=1.83 (1.37 2.26)). Significant linkage disequilibrium was observed between the 105A/C and 137G/C polymorphisms in the 5' flanking region of the IL-18 gene (D=0.58, P<0.0001). However, in asthmatic patients the 105A allele was not associated with either total serum IgE or IL-18 levels. CONCLUSION: The 105A/C polymorphism of the IL-18 gene may be associated with the pathogenesis of asthma. PMID- 12911785 TI - Promoter polymorphism in the 5-lipoxygenase (ALOX5) and 5-lipoxygenase-activating protein (ALOX5AP) genes and asthma susceptibility in a Caucasian population. AB - BACKGROUND: 5-Lipoxygenase (5-LO) and 5-lipoxygenase-activating protein (FLAP) are essential for cysteinyl-leukotriene (cys-LT) production, critical mediators in asthma. OBJECTIVE: We sought to identify novel promoter polymorphisms within the FLAP (ALOX5AP) gene promoter and test the role of these and the previously identified 5-LO (ALOX5) Sp1 promoter polymorphism in asthma susceptibility. METHODS: To assess genetic association with asthma phenotypes, we genotyped 341 Caucasian families (containing two asthmatic siblings) and non-asthmatic control subjects (n=184). Genetic association was determined by case-control and transmission disequilibrium test (TDT) analyses. To determine the functional role of polymorphisms on basal transcription, we generated ALOX5AP-promoter-luciferase constructs and transiently transfected human HeLa cells. RESULTS: A novel G/A substitution at -336 bp and a poly(A) repeat (n=19 or 23) at position -169 to 146 bp were identified in the ALOX5AP promoter. Genotyping found the -336 A and poly(A19) alleles at frequencies of q=0.06 and 0.12, respectively. No ALOX5AP allele was associated with asthma or asthma-related phenotypes in case-control or TDT analyses. ALOX5AP-promoter-luciferase analyses did not support a functional role of the -336 or poly(A) polymorphism in determining basal transcription. The ALOX5 Sp1 polymorphism was predominantly homozygous wild-type 5/5 (frequency q=0.70) and heterozygous 4/5 (q=0.23) genotypes and no allele was associated with asthma or asthma-related phenotypes. CONCLUSION: Taken together, these data do not support a significant role for these polymorphisms in genetic susceptibility to asthma in the Caucasian population. PMID- 12911786 TI - Polymorphisms in the interleukin-4 and interleukin-4 receptor alpha chain genes confer susceptibility to asthma and atopy in a Caucasian population. AB - BACKGROUND: IL-4 by binding to its receptor (IL-4R) is essential for the development of airway inflammation present in asthma, through the induction of IgE synthesis in B cells and differentiation of T cells to a Th2 phenotype. OBJECTIVE: To investigate the role of four common polymorphisms in the IL-4 (IL4 34CT and IL4-589CT) and IL-4Ralpha chain (IL4RAI50V and IL4RAQ576R) genes in conferring susceptibility to the development of atopy and/or asthma. METHODS: Two polymorphisms in the IL-4 gene promoter, IL4-34CT and IL4-589CT, and two polymorphisms in the IL-4Ralpha chain gene, IL4RAI50V and IL4RAQ576R, have been genotyped using PCR-based methods in 341 asthmatic families and in 184 non asthmatic adults recruited from the south of England. RESULTS: Case-control analysis did not reveal differences in the distribution of the four polymorphisms between asthmatics and controls. However, the transmission disequilibrium test showed that the IL4-589 T allele was preferentially transmitted to asthmatic children (P=0.036) and that the IL4RAQ576 was preferentially transmitted to children with atopic asthma (P=0.018). Haplotype analysis showed a strong association between the IL4-34T/-589T haplotype and asthma per se (P=0.041), and a strong association between the IL4RA I50/Q576 haplotype and atopic asthma (P=0.006). CONCLUSION: Our data suggest that polymorphisms in the IL-4 and IL 4Ralpha chain genes might play a role both conferring susceptibility to and modulating severity of atopy and asthma. PMID- 12911787 TI - Expression of CS-1 fibronectin precedes monocyte chemoattractant protein-1 production during elicitation of allergic contact dermatitis. AB - BACKGROUND: Leucocyte migration within inflammatory skin compartments in allergic contact dermatitis (ACD) is the result of a sophisticated multi-step event where multiple molecules are involved. OBJECTIVE: Since non-antigen-specific mechanisms have been described as an early participant in elicitation of ACD, we investigated the kinetics of the expression of monocyte chemoattractant protein-1 (MCP-1/CCL2) and the type of infiltrating cells. We compared the time course production of MCP-1/CCL2 with connecting segment-1 (CS-1) fibronectin and thymus and activation-regulated chemokine (TARC/ CCL17) expression. METHODS: Biopsies from 10 individuals challenged in their back with the antigen responsible for their contact dermatitis and an irrelevant antigen were taken at different times and histology, immunohistochemistry for CS-1 fibronectin, TARC/CCL17, CD3, CD68, CXCR3, CCR4 and in situ hybridization for MCP-1/CCL2 were performed. RESULTS: At positive antigen stimulated sites expression of MCP-1/CCL2 by basal keratinocytes and isolated cells in dermis started at 10 h. CS-1 fibronectin and TARC/CCL17 expression by blood endothelial cells was found at 2 and 10 h, respectively. This was followed by dermal accumulation of mononuclear cells with a significant increase of CD3+ and CD68+cells. At 48 h, approximately 58% of infiltrating cells were CXCR3+, and 35% CCR4+. CONCLUSIONS: We showed evidence of the fact that CS-1 fibronectin expression precedes the production of MCP-1/CCL2 and TARC/CCL17 in the skin of patients with ACD, suggesting that these molecules participate in the early complex process of migrating mononuclear cells during elicitation of ACD. PMID- 12911788 TI - Regulation and kinetics of platelet-activating factor and leukotriene C4 synthesis by activated human basophils. AB - BACKGROUND: Allergic disease is the result of an interplay of many different cell types, including basophils and mast cells, in combination with various inflammatory lipid mediators, such as platelet-activating factor (PAF) and leukotrienes (LT). LTC4 synthesis by human basophils has been studied quite extensively. However, not much is known about the synthesis of PAF by human basophils. OBJECTIVE: In this study, we have made a comprehensive comparison between the kinetics of PAF and LTC4 synthesis, in highly purified basophils, activated with different stimuli or with combinations of stimuli. METHODS: Synthesis of PAF and LTC4 by human basophils was determined with commercially available assay kits. The basophils were activated with C5a, fMLP, PMA, allergen or anti-IgE, in the absence and presence of IL-3 and/or in combination with elevation of cytosolic free Ca2+ by the sarcoplasmic reticulum Ca2+-ATPase inhibitor thapsigargin. RESULTS: Most stimuli were found to induce both PAF and LTC4 synthesis. PAF synthesis and LTC4 release were enhanced by preincubation of the basophils with IL-3 or by elevation of cytosolic free Ca2+ by thapsigargin. Incubation of human basophils with IL-3 alone or thapsigargin alone did not result in detectable synthesis of PAF and LTC4, whereas the combination of the two resulted in high amounts of PAF and LTC4 synthesis. Depending on the stimulus used, LTC4 release was 5-100-fold higher than PAF synthesis. In addition, PAF, but not LTC4, was transiently detected, probably due to PAF degradation. LTC4 and PAF synthesis was strongly blocked by inhibitors of cytosolic phospholipase A2, indicating that this enzyme is involved in PAF and LTC4 synthesis by activated human basophils. CONCLUSION: This study provides a first comprehensive comparison of PAF and LTC4 synthesis in highly purified human basophils, stimulated with a variety of stimuli. PMID- 12911789 TI - Activation of mast cells by immunoglobulin E-receptor cross-linkage, but not through adenosine receptors, induces A1 expression and promotes survival. AB - BACKGROUND: Mast cells are a potent source of mediators that regulate the inflammatory response in allergy and asthma. Mast cells can be activated through different receptors, for example, via cross-linkage of the high-affinity IgE receptor (Fc epsilon RI) and by adenosine acting on specific receptors. We have recently described mast cell survival of an IgE receptor activation by up regulation of the anti-apoptotic gene A1. OBJECTIVE: To compare mast cell survival and expression of A1 after activation through the Fc epsilon RI and by an adenosine agonist. METHODS: Bone marrow-derived, cultured mouse mast cells (BMCMC) were activated either with IgE+antigen or with the adenosine receptor agonist 5'-N-ethylcarboxamido adenosine (NECA). Release of beta-hexosaminidase, cell viability, phosphorylation of Akt and IkB-alpha, and expression of pro survival and pro-apoptotic genes were measured after activation. RESULTS: Activation of BMCMC with NECA caused the release of beta-hexosaminidase, although to a lesser extent than after Fc epsilon RI activation (33% and 98%, respectively). Activation by both NECA and Fc epsilon RI stimulated phosphorylation of Akt (Ser473 and Thr308) and IkB-alpha (Ser32), both of which are implicated in the regulation of cell survival. However, only cells that were activated through Fc epsilon RI, but not by NECA, expressed A1 and exhibited an increased survival rate compared to the control. CONCLUSION: These results show that adenosine receptor activation of BMCMC does not induce the same survival programme in mast cells as does activation through Fc epsilon RI. These findings may be important for understanding the role that mast cells play in asthma provoked by different stimuli. PMID- 12911790 TI - Increased expression of surface activation markers on neutrophils following migration into the nasal lumen. AB - BACKGROUND: The sequence of events following the recruitment of a free-flowing neutrophil in the peripheral circulation, via adhesion, migration and release of mediators, to a neutrophil on the surface of the nasal epithelium is a co ordinated process. Little is known about the state of neutrophil activation following this course of events. OBJECTIVES: To investigate the expression of surface activation markers on neutrophils, reflecting activation during their recruitment to the nose, and to see whether the inflammatory process during allergic rhinitis influences this process. METHOD: Nine healthy controls and 12 patients with grass pollen-induced intermittent allergic rhinitis were investigated during the peak of the pollen season. The expression of CD11b, CD66b and CD63 on the neutrophil cell surface, as a reflection of activation, was analysed using flow cytometry. Neutrophils were derived from peripheral blood and nasal lavage fluid. In addition, eosinophil cationic protein (ECP) and myeloperoxidase (MPO) as well as L-, P- and E-selectins in the nasal lavage fluid were analysed using RIA and ELISA, respectively. RESULTS: A marked increase in the expression of all three CD markers on the neutrophil cell surface was noticed following migration from the bloodstream to the surface of the nasal mucosa. At the peak of the grass pollen season, the MPO levels increased, reflecting an increase in the total number of nasal fluid neutrophils. In parallel, the expression of CD11b was further augmented. The expression of the CDb11b was reduced on neutrophils remaining in the circulation. In addition, the level of L selectin was reduced on neutrophils derived from the blood during allergic inflammation. CONCLUSION: Neutrophils might become activated during their transfer from the blood to the surface of the nasal mucosa, but these changes may also be due to depletion of activated neutrophils in the blood via activated endothelial/epithelial adhesion and chemoattractant measures. The increased expression of surface activation markers during allergic rhinitis suggests roles for neutrophils in the inflammatory process. PMID- 12911791 TI - In vitro stability and immunoreactivity of the native and recombinant plant food 2S albumins Ber e 1 and SFA-8. AB - BACKGROUND: The ability of an intact protein to reach the circulatory system may be a prerequisite to allergenicity and many allergens, particularly those from plant foods, have been found to be consistently more resistant to digestion by pepsin than other proteins. OBJECTIVE: This study assessed the pepsinolytic stability of native 2S albumins from Brazil nut and sunflower seed and their recombinant versions produced in Pichia pastoris. The physicochemical stability of native and recombinant Brazil nut 2S albumins and recombinant sunflower seed 2S albumin was also assessed. The immunoreactivity of native Brazil nut 2S albumin and recombinant 2S albumins was compared using serum from patients allergic to Brazil nuts and animals immunized with native 2S albumins. METHODS: Digestibility was measured in simulated gastric fluid followed by SDS-PAGE. Circular dichroism spectra were used to analyse unfolding, as proteins were denatured by temperature, pH and guanidinium chloride. Immunoreactivity was assessed by immunoblot, RAST and ELISA. RESULTS: Brazil nut 2S albumin was significantly more resistant to proteolytic digestion than other Brazil nut proteins. It was also resistant to thermally and chemically induced denaturation. Equally high resistance to proteolytic digestion was observed with sunflower seed 2S albumin. The recombinant albumins mirrored their native counterparts in stability and immunoreactivity. CONCLUSION: The important food allergen Brazil nut 2S albumin is as stable to digestion as is sunflower seed 2S albumin, whose allergenicity has yet to be determined. The 2S albumins and their recombinant counterparts could not be easily denatured by physicochemical treatments. The results suggest that 2S albumin is the only Brazil nut protein to reach the gut immune system intact. The production of properly folded recombinant proteins will facilitate mechanistic studies as well as diagnostic testing and antigen-based therapies. PMID- 12911792 TI - In vitro and in vivo allergenicity of recombinant Bet v 1 compared to the reactivity of natural birch pollen extract. AB - BACKGROUND: Diagnostic procedures using natural extracts show only limited quantitative correlation between in vivo and in vitro results. Highly pure recombinant allergens might show more predictive findings. OBJECTIVE: The aim of this study was to compare natural birch pollen extract (BPE) and recombinant Betula verrucosa (rBet v 1) for their diagnostic value comparing skin prick tests (SPTS) and nasal provocation tests (NPTS) with specific IgE in the serum. METHODS: Thirty-four patients allergic to birch pollen and five healthy controls were investigated. SPT and NPT were performed with BPE and rBet v 1 at different concentrations. Specific serum IgE was measured by the Pharmacia CAP system. RESULTS: Commercial BPE and rBet v 1 (10 micro g/mL) were able to elicit similar allergenic reactions in vivo and IgE binding in vitro. SPT reflects immediate type allergy as determined by NPT to a higher degree than specific IgE, for both reagents. To cause allergic reactions in NPT, higher amounts of rBet v 1 were needed than for skin tests and the sensitivity was lower than with BPE. CONCLUSION: rBet v 1 alone is sufficient for a reliable diagnosis of birch pollen allergy in most patients and induces comparable skin test reactivity as BPE, but less allergic reactions in nasal provocations. PMID- 12911793 TI - Proteolytic activity of the house dust mite allergen Der p 1 enhances allergenicity in a mouse inhalation model. AB - BACKGROUND: We have recently demonstrated that intraperitoneal immunization of mice with proteolytically active Der p 1, the major house dust mite allergen, results in a significant and selective enhancement of total and Der p 1-specific IgE synthesis compared to mice immunized with proteolytically inactive Der p 1. OBJECTIVE: To investigate whether the proteolytic activity of Der p 1 would lead to enhanced inflammatory cellular infiltration of the lungs and systemic IgE production when administered through the respiratory system, which is the natural route of entry for this allergen. METHODS: Groups of mice were initially sensitized with proteolytically active Der p 1 through the intraperitoneal and the subcutaneous routes and subsequently exposed intranasally to either proteolytically active Der p 1, inactive Der p 1 or PBS. The extent of cellular infiltration of the lungs and systemic IgE production in the three animal groups were then compared. RESULTS: Here, we show for the first time that the administration of proteolytically active Der p 1 to mice through the intranasal route leads to significant inflammatory cellular infiltration of the lungs and systemic production of IgE. CONCLUSIONS: These data underline the important role of the proteolytic activity of Der p 1 in driving the allergic response in the lungs. PMID- 12911794 TI - The role of the mast cell in asthma. PMID- 12911796 TI - The push against vaginal birth. PMID- 12911797 TI - Does postpartum length of stay affect breastfeeding duration? A population-based study. AB - BACKGROUND: Short postpartum hospital stays may leave inadequate time for women to receive assistance with breastfeeding. Women leaving the hospital early may also have household responsibilities that could interfere with breastfeeding. This study examined the relationship between postpartum length of stay and breastfeeding cessation. METHODS: This study used data from 10,519 respondents to the California Maternal and Infant Health Assessment (MIHA) surveys from 1999 to 2001. MIHA is an annual statewide stratified random sample, population-based study of childbearing women in California. Survival analysis was used to examine the relationship between length of stay and length of time breastfeeding. Women were asked about the number of nights their infant stayed in the hospital at birth, whether they breastfed, and if so, the age of the child when they stopped. Hospital stay was defined in three categories: standard (2 nights for a vaginal delivery, 4 nights for a cesarean section), or shorter or longer than the standard stay. RESULTS: Approximately 88 percent of women initiated breastfeeding. Unadjusted predictors of breastfeeding cessation included short or long postpartum stay; young maternal age; Hispanic, African American, or Asian/Pacific Islander race/ethnicity; being unmarried; low income or education level; primiparity; being born in the 50 United States or the District of Columbia; smoking during pregnancy; and low infant birthweight. After adjustment for potential confounders, women with a short stay remained slightly more likely to terminate breastfeeding than women with a standard stay (relative risk, 1.11, 95% confidence interval 1.01, 1.23). CONCLUSION: Women who leave the hospital earlier than the standard recommended stay are at somewhat increased risk of terminating breastfeeding early. PMID- 12911798 TI - Evaluating innovations in maternity care: methodological approaches to a baseline postal survey. AB - BACKGROUND: A network of four public maternity hospitals in Melbourne, Australia, has implemented a range of initiatives aimed at improving maternity care. Comprehensive evaluation aims to determine whether or not the maternity service enhancement strategies have led to improvements in women's views and experiences of care. This paper discusses the baseline survey study design and methodological issues associated with the recruitment and reminder processes. METHOD: All women who gave birth over a 14-week period in 1999 at one of the four maternity units, except those who had a stillbirth or neonatal death, were invited to participate in the baseline postal survey. Questionnaires were sent to women at 3 months postpartum. A system of written and telephone reminders was instituted. RESULTS: The overall response fraction to the baseline survey was 65.2 percent (1256/1922). The sample was representative in terms of maternal age, method of birth, and infant birthweight. Women born overseas of non-English speaking background, single women, and women having their second or subsequent baby were underrepresented. CONCLUSIONS: The pleasing response fraction in part may be attributed to the method of recruitment and the reminder processes. Rigorous and careful approaches to methodology enhance study integrity and provide context to the interpretation of study findings. PMID- 12911799 TI - Labor induction rate variation in upstate New York: what is the difference? AB - BACKGROUND: Labor induction rates in the United States rose from 9.0 percent in 1989 to 20.5 percent in 2001, but reasons for the increase are poorly defined. A birth database from a region of upstate New York, including 31,352 deliveries from 1998 through 1999, was used to determine the degree of variation of labor induction rates among hospitals and practitioners. METHODS: Total and elective labor induction rates were calculated for 16 hospitals and individual staff, and then evaluated using chi-square testing and regression. RESULTS: Using all laboring women as the denominator, the regional labor induction rate was 20.8 percent; of these inductions, 25 percent had no apparent medical indication. Total induction rates and percent of elective inductions that were elective varied significantly among hospitals (10%-39% and 12%-55%, respectively, p<0.0001) and among practitioners within hospitals (7%-48% and 3%-76%, respectively, p<0.0001). Hospitals varied in size, risk status, and cesarean section rates, but these factors did not correlate with induction rates. CONCLUSIONS: Labor induction rates are highly variable among and within hospitals. Delivery volume, population risk status, and differences in cesarean section rates did not explain this variation. PMID- 12911800 TI - Impact of postnatal depression on breastfeeding duration. AB - BACKGROUND: Postnatal depression can cause adverse effects on both mother and infant, but its impact on breastfeeding duration is poorly understood. The aim of this study was to investigate the relationship between maternal postnatal depression and breastfeeding duration. METHODS: A cohort of 1745 women was recruited on the postnatal wards of two large Australian obstetric hospitals. Self-report questionnaires were completed at recruitment, and at 2, 6, and 12 months postpartum. Breastfeeding status was determined at each follow-up, and the Edinburgh Postnatal Depression Scale was used to screen for symptoms of depression. Diagnostic psychological interviews were conducted on a subsample of women at each interval. RESULTS: Breastfeeding was initiated by 96 percent of the participants; at 2 months 79 percent were still breastfeeding, 57 percent at 6 months, and 22 percent at 12 months. Of the 18 percent of participants diagnosed with postnatal depression, the onset occurred before 2 months in 63 percent of cases. Median duration of breastfeeding was 26 weeks for women with early-onset depression, 28 weeks for women with late-onset depression, and 39 weeks for women without depression. After adjustment for confounding factors, early cessation of breastfeeding was found to be significantly associated with postnatal depression (adjusted hazard ratio 1.25, 95% CI 1.03-1.52). Onset of postnatal depression occurred before cessation of breastfeeding in most cases. CONCLUSIONS: Postnatal depression has a significant negative impact on breastfeeding duration. Assistance with breastfeeding issues should be included in the management of postnatal depression. PMID- 12911801 TI - Breastfeeding, bed-sharing, and infant sleep. AB - BACKGROUND: Expectations for infant sleep development and for the appropriate degree of parental proximity for infant sleep are culturally weighted and historically shifting aspects of parenting behavior, and are known to affect breastfeeding prevalence and duration. This paper examined how new parents managed night-time feeding in the first 4 months, with a particular focus on the relationship between breastfeeding, infant sleep location, and sleep bout duration. METHODS: Sleep logs and semistructured interviews were used with a sample of 253 families in North Tees, United Kingdom, to explore how parents responded to their infant's sleep patterns, how breastfeeding parents managed night-time feeding, and whether bed-sharing was a common strategy. RESULTS: A clear relationship between breastfeeding and parent-infant bed-sharing was demonstrated. Some evidence indicated that bed-sharing may promote breastfeeding. CONCLUSIONS: An understanding of the role of infant feeding practice on infant sleep and parental caregiving at night is a crucial element in breastfeeding promotion and enhancement of infant health. Health professionals should discuss safe bed-sharing practices with all parents. PMID- 12911802 TI - Selected coping strategies in labor: an investigation of women's experiences. AB - BACKGROUND: Antenatal education classes offer women information about labor and birth and ways of coping with pain and emotional distress. The purpose of this paper was to describe women's experiences of using, starting, and discontinuing three coping strategies in labor that were taught in antenatal education classes. METHODS: An exploratory research design was used in which 121 women were interviewed within 72 hours of the birth of their first child. Information was obtained on why women initiated and discontinued their use of three coping strategies (breathing technique, postural changes, relaxation technique) and the reported effects of use. RESULTS: The effects of the coping strategies investigated varied widely among participants. Common aspects of care, changes of environment, and use of pharmacological pain relief affected women's discontinuation of coping strategies. CONCLUSIONS: The implications of study findings for clinical practice include the need for caregivers to provide women with accurate information about the effects of coping strategies and to be alert to aspects of care that may disrupt women's use of strategies. PMID- 12911803 TI - Breastfeeding patterns of primiparous mothers in Hong Kong. AB - BACKGROUND: An understanding of patterns of breastfeeding is necessary for the effective implementation of breastfeeding promotion and intervention programs. In Hong Kong, little current data have been gathered on women's breastfeeding rates. The objective of this study was to determine how patterns of breastfeeding, maternal demographics, and maternal employment affect continuation of breastfeeding in primiparous women in Hong Kong. METHOD: A longitudinal self report survey was used to collect data when first-time mothers (n=218) were in the hospital, at 1, 3, 6, 9, and 12 months postpartum, or until they weaned their infant. All data (self-report survey, demographic data, and follow-up telephone surveys) were collected in Cantonese and then translated into English. Data were analyzed by determining, first, the influence of individual variables on the length of breastfeeding using a simple Cox regression analysis, and second, by grouping variables according to time sequence and entering them into a Cox regression model in 4 sequential phases. RESULTS: Factors that were significantly associated with continuation of breastfeeding were maternal age (HR=0.97; p=0.048); attendance at a prenatal breastfeeding class (HR=0.69; p=0.020); intended weeks of breastfeeding (HR=0.97; p<0.001); breastfeeding score in hospital (HR=0.99; p=0.009); and length of exclusive breastfeeding (HR=0.93; p<0.001). Similar results were obtained in the multiphase Cox regression analysis; only the breastfeeding score in hospital became marginally insignificant (p=0.053) after adjusting for demographics, prenatal, and other immediate postpartum factors. CONCLUSIONS: Short periods of exclusive breastfeeding and early supplementation were common in this sample. Unlike previous research, maternal employment was not a statistically significant factor in length of continued breastfeeding. Study findings show that multiple factors influence continued breastfeeding in Hong Kong, suggesting further areas for investigation. Changes in practice may improve continued and exclusive breastfeeding rates. PMID- 12911804 TI - Sheila Kitzinger's letter from Europe: the politics of birth. PMID- 12911805 TI - Early skin-to-skin contact for mothers and their healthy newborn infants. PMID- 12911809 TI - Impact of an educational intervention on breastfeeding. PMID- 12911811 TI - ISBT - Guidelines for validation and maintaining the validation state of automated systems in blood banking. PMID- 12911812 TI - Toxoplasma gondii: perfecting an intracellular life style. AB - Toxoplasma gondii is a widespread protozoan parasite that infects all nucleated cell types of warm-blooded vertebrates. Parasite motility is regulated by polymerization of new actin filaments that provide a substrate for the small myosin TgMyoA. Interaction between the cytoplasmic tails of parasite adhesins and the actin-binding protein aldolase links these cell surface proteins with the cytoskeleton. Translocation of adhesins coupled to extracellular receptors allows the parasite to glide across the substrate. This conserved system is important for active penetration into host cells and tissue migration by T. gondii. Entry into the host cell is accompanied by dramatic remodeling of the intracellular vacuole that the parasite resides in. This compartment resists fusion with host cell endocytic organelles, yet recruits mitochondria and endoplasmic reticulum in order to gain access to host cell nutrients. The combined abilities to actively penetrate host cells and control the fate of the parasite-containing vacuole contributes to the remarkable success of T. gondii as an intracellular parasite. PMID- 12911813 TI - Taking possession: biogenesis of the Salmonella-containing vacuole. AB - The Gram-negative pathogen Salmonella enterica can survive and replicate within a variety of mammalian cells. Regardless of the cell type, internalized bacteria survive and replicate within the Salmonella-containing vacuole, the biogenesis of which is dependent on bacterially encoded virulence factors. In particular, Type III secretion systems translocate bacterial effector proteins into the eukaryotic cell where they can specifically interact with a variety of targets. Salmonella has two distinct Type III secretion systems that are believed to have completely different functions. The SPI2 system is induced intracellularly and is required for intracellular survival in macrophages; it plays no role in invasion but is categorized as being required for Salmonella-containing vacuole biogenesis. In contrast, the SPI1 Type III secretion system is induced extracellularly and is essential for invasion of nonphagocytic cells. Its role in post-invasion processes has not been well studied. Recent studies indicate that Salmonella containing vacuole biogenesis may be more dependent on SPI1 than previously believed. Other non-SPI2 virulence factors and the host cell itself may play critical roles in determining the intracellular environment of this facultative intracellular pathogen. In this review we discuss the recent advances in determining the mechanisms by which Salmonella regulate Salmonella-containing vacuole biogenesis and the implications of these findings. PMID- 12911814 TI - Mycobacterium tuberculosis phagosome maturation arrest: selective targeting of PI3P-dependent membrane trafficking. AB - The ability of Mycobacterium tuberculosis to enter host macrophages, and reside in a phagosome, which does not mature into a phagolysosome, is central to the spread of tuberculosis and the associated pandemic involving billions of people worldwide. Tuberculosis can be viewed as a disease with a significant intracellular trafficking and organellar biogenesis component. Current understanding of the block in M. tuberculosis phagosome maturation also sheds light on fundamental aspects of phagolysosome biogenesis. The maturation block involves interference with the recruitment and function of rabs, rab effectors (phosphatidylinositol 3-kinases and tethering molecules such as EEA1), SNAREs (Syntaxin 6 and cellubrevin) and Ca2+/calmodulin signaling. M. tuberculosis analogs of mammalian phosphatidylinositols interfere with these systems and associated processes. PMID- 12911815 TI - Yeast functional analysis: identification of two essential genes involved in ER to Golgi trafficking. AB - We screened for genes potentially involved in the secretory and vacuolar pathways a collection of 61 yeast strains, each bearing an essential orphan gene regulated by the tetO7-CYC1 promoter that can be down-regulated by doxycycline. After down regulating the expression of these genes, we performed systematic Western blot analysis for markers of the secretory and vacuolar pathways that undergo post translational modifications in their intracellular trafficking. Accumulation of protein precursors, revealed by Western immunoblot analysis, indicates defects in the secretory pathway or in associated biochemical modifications. After screening the whole collection, we identified two genes involved in ER to Golgi trafficking: RER2, a cis-prenyl transferase, and USE1, the function of which was unknown. We demonstrated that repression of USE1 also leads to BiP secretion, and therefore likely affects retrograde, in addition to anterograde, ER to Golgi trafficking. The collection also includes two essential genes involved in intracellular trafficking that were conveniently repressed without resulting growth or trafficking defects. PMID- 12911816 TI - Regulation of EGF-induced phospholipase C-gamma1 translocation and activation by its SH2 and PH domains. AB - Translocation of phospholipase C-gamma1 is essential for its function in response to growth factors. However, in spite of recent progress, the phospholipase C gamma1 translocation pattern and the molecular mechanism of the translocation are far from fully understood. Contradictory results were reported as to which domain, PH or SH2, controls the epidermal growth factor-induced translocation of phospholipase C-gamma1. In this communication, we studied epidermal growth factor induced translocation of phospholipase C-gamma1 by using comprehensive approaches including biochemistry, indirect fluorescence and live fluorescence imaging. We provided original evidence demonstrating that: (i) endogenous phospholipase C gamma1, similar to YFP-tagged phospholipase C-gamma1, translocated to endosomes following its initial translocation from cytosol to the plasma membrane in response to epidermal growth factor; (ii) phospholipase C-gamma1 remained phosphorylated in endosomes, but phospholipase C-gamma1 activity is not required for its translocation, which suggests a signaling role for phospholipase C-gamma1 in endosomes; (iii) the PH domain was not required for the initial translocation of phospholipase C-gamma1 from cytosol to the plasma membrane, but it stabilizes phospholipase C-gamma1 in the membrane at a later time; (iv) the function of the phospholipase C-gamma1 PH domain in stabilizing phospholipase C-gamma1 membrane association is very important in maintaining the activity of phospholipase C gamma1; and (v) the role of the PH domain in phospholipase C-gamma1 membrane association and activation is dependent on PI3K activity. We conclude that the phospholipase C-gamma1 SH2 and PH domains coordinate to determine epidermal growth factor-induced translocation and activation of phospholipase C-gamma1. PMID- 12911817 TI - Persistence of Golgi matrix distribution exhibits the same dependence on Sar1p activity as a Golgi glycosyltransferase. AB - We investigated the relative distributional persistence of Golgi "matrix" proteins and glycosyltransferases to an endoplasmic reticulum exit block induced by expression of a GDP-restricted Sar1p. HeLa cells were microinjected with plasmid encoding the GDP-restricted mutant (T39N) of Sar1p to block endoplasmic reticulum exit and then scored for the distribution of GM130 (Golgi matrix protein of 130 kDa), a cis located golgin; p27, a member of the p24 family of proteins; giantin, a protein that interacts indirectly with GM130; and the Golgi glycosyltransferase, N-acetylgalactosaminyltransferase-2 (GalNAcT2). All of these proteins lost their compact, juxtanuclear distribution and displayed characteristics of endoplasmic reticulum/cytoplasmic accumulation with the same dependence on plasmid concentration. The kinetics of redistribution of GM130 and GalNAcT2 were identical. Expression of Sar1pT39N displaced the COPII coat protein Sec13p from endoplasmic reticulum exit sites consistent with disruption of these sites. This occurred without disturbing the overall distribution of endoplasmic reticulum membrane. Furthermore, the reassembly of a juxtanuclear Golgi matrix as assayed by the distribution of GM130 following washout of the Golgi disrupting drug, brefeldin A, was blocked by microinjected Sar1pT39N plasmids. We conclude that the persistence, i.e. stability and maintenance, of Golgi matrix distribution and its reassembly following drug disruption are exquisitely dependent on Sar1p activity. PMID- 12911818 TI - Carboxyl-terminal truncations of human anion exchanger impair its trafficking to the plasma membrane. AB - The human anion exchanger AE1 (Band 3) is an abundant glycoprotein localized in plasma membrane of red cells and is responsible for the electro-neutral exchange of chloride for bicarbonate. In order to determine the role of the carboxyl terminal tail of AE1 in its expression, function and trafficking to the plasma membrane, we generated a series of five constructs encoding truncation mutants missing the last 5 (Delta5), 11 (Delta11), 15 (Delta15), 20 (Delta20) or 35 (Delta35) amino-acids. In transiently transfected HEK 293 cells, immunoblotting of whole cell extracts showed that all the proteins were expressed at the same level as full-length AE1, except Delta20 and particularly Delta35, which showed a reduced expression. Furthermore, the last 15 amino-acids were not required for AE1 folding in the membrane, since Delta5, Delta11 and Delta15 were able to bind to an inhibitor affinity matrix, while Delta20 and Delta35 exhibited poor binding. Immunofluorescence and deglycosylation results showed that Delta15 and Delta11 were retained intracellularly, whereas a lower amount of Delta5 compared with WT trafficked to the plasma membrane. These results indicate that an intact C-terminal tail of human AE1 is important for efficient AE1 trafficking to the plasma membrane. PMID- 12911822 TI - The loss of retinoic acid receptor alpha, beta and alcohol dehydrogenase3 expression in non-small cell lung cancer. AB - OBJECTIVE: The reason for unsuccessful retinoid chemoprevention of lung cancer is obscure. Therefore we examined the retinoid-related receptor expression and the tissue distribution as well as the possible interaction between retinoid-related receptor expression and cigarette smoke. METHODOLOGY: Using semiquantitative conventional reverse transcriptase polymerase chain reaction (RT-PCR), quantitative real-time RT-PCR, in situ hybridization and immunostaining, we evaluated the expression and the distribution of retinoic acid receptor alpha, beta, gamma, retinoid X receptor alpha, beta, gamma, thyroid hormone receptor alpha, beta, and alcohol dehydrogenase3 (a well-characterized retinoic acid responsive gene) in malignant and non-malignant tissues obtained from patients with non-small cell lung cancer. We also assessed the response of the bronchial epithelial cell line, BEAS-2B, following exposure to cigarette smoke extract. RESULTS: Compared with non-malignant tissues, malignant tissues had a 45% (P < 0.02) reducion in retinoic acid receptor alpha and beta expression, while other receptors were not affected. The expression of the alcohol dehydrogenase3 gene in malignant tissue was 50% of that observed in non-malignant tissue (P < 0.05). There was a significant correlation between the retinoic acid receptor beta-mRNA level and the alcohol dehydrogenase3-mRNA level (r = 0.52, P < 0.05). In situ hybridization and immunostaining demonstrated that retinoic acid receptor alpha and beta were localized in the airway and alveolar epithelial cells and their expression was diminished to 30% in malignant tissues compared to non-malignant tissues (P < 0.05). Interestingly, cigarette smoke extract decreased retinoic acid receptor beta mRNA in BEAS-2B cells (P < 0.05). CONCLUSIONS: The present findings suggest a possible association of the loss of retinoic acid receptor alpha, beta and alcohol dehydrogenase3 with lung carcinogenesis. PMID- 12911823 TI - Modulation of adenovirus infection in vitro by antisense oligodeoxynucleotides. AB - OBJECTIVE: Antisense oligodeoxynucleotides (ODNs) may represent a novel, airway directed approach to the treatment of adenovirus infection of the lung, for which no specific therapy exists. This study assessed the efficacy of antisense ODNs in modulating adenovirus infection in vitro. METHODOLOGY: A biological assay, which quantified viral plaque formation by wild type adenovirus 5 in a lung epithelial cell line (A549), was used to evaluate the inhibitory effect of a number of antisense ODNs targeted to the early (E) 1 A and protein IX genes of adenovirus 5. Antisense ODNs (20-21mers, phosphorothioate end-protected) were designed to straddle the initiation of translation (AUG) codon of the mRNA of the targeted gene. RESULTS: There was a consistent and significant (P < 0.005) reduction in viral plaque formation in those cells treated with an E1A antisense ODN, compared with the nonsense control ODN. Neither the addition of a cationic lipid (Lipofectamine), nor increasing the concentration of ODN from 1 micro mol to 15 micro mol enhanced the original inhibitory effect observed with the E1A antisense ODN. CONCLUSIONS: An antisense ODN targeted to the E1A gene can specifically inhibit adenovirus 5 infection in vitro, suggesting a potential therapeutic role for antisense ODNs in adenovirus infection of the lung. PMID- 12911824 TI - Effect of the proteolytic enzyme serrapeptase in patients with chronic airway disease. AB - OBJECTIVES: The proteolytic enzyme serrapeptase (SER) is widely used in clinical practice in Japan. We investigated the effect of SER on sputum properties and symptoms in patients with chronic airway diseases. METHODS: This study was an open-labelled trial with a non-treatment control group. Patients were randomly assigned to oral treatment with (n = 15) and without (n = 14) SER 30 mg/day for 4 weeks. Patients collected sputum samples for about 4 h in the morning on the day the trial began and 4 weeks later. We measured the amount of sputum by weighing. Part of each sputum sample was weighed and then completely dried and reweighed. The percentage solid component, viscosity and elasticity of the sputum were measured. Mucociliary transportability index was measured using ciliated bovine trachea ex vivo. Sputum smears were also prepared to count sputum neutrophils. Patients' symptoms were assessed by a questionnaire that used a visual analogue scale. RESULTS: After 4 weeks of SER treatment, sputum weight in the morning, percentage solid component, viscosity and elasticity of sputum, sputum neutrophil count, frequency of coughing and frequency of expectoration significantly decreased. The mean mucociliary transportability index increased from 13.3 +/- 1.8 to 24.4 +/- 2.5 (P = 0.0103). CONCLUSIONS: SER may exert a beneficial effect on mucus clearance by reducing neutrophil numbers and altering the viscoelasticity of sputum in patients with chronic airway diseases. PMID- 12911825 TI - Transforming growth factor beta-1 level in pleural effusion. AB - OBJECTIVE: Transforming growth factor-beta1 is an important immunomodulator. The diagnostic role of TGF-beta1 has not been systematically investigated in pleural effusion. METHODOLOGY: A prospective clinical study of 45 patients (23 men, 22 women; mean age 49 +/- 21 years) with pleural effusion was performed. Of these patients, 19 had malignant pleural effusion, 14 had tuberculous pleural effusion, seven had empyema/parapneumonic pleural effusion, and five had transudative pleural effusion due to congestive heart failure. The concentrations of TGF-beta1 were measured by ELISA in all pleural fluid samples and in serum samples only from patients with malignant and tuberculous pleural effusions. RESULTS: The median TGF-beta1 levels of malignant, tuberculous and empyema/parapneumonic pleural effusions were 7.25 ng/mL, 7.81 ng/mL, and 9.75 ng/mL, respectively. There was no significant difference between them. The median TGF-beta1 level was 5.62 ng/mL in the transudate pleural effusion group and it was significantly lower than that in the empyema/parapneumonic group (P < 0.05). The pleural fluid TGF-beta1 levels did not correlate with cell profiles of the pleural fluid. The median serum TGF-beta1 levels in malignant and tuberculous pleural effusion groups were 7.38 ng/mL and 7.38 ng/mL, respectively. There was no significant difference between the levels of TGF-beta1 in paired samples of serum and pleural fluid. CONCLUSIONS: This study shows that TGF-beta1 concentrations in exudative pleural effusions are higher than those in transudative effusions secondary to congestive heart failure but TGF-beta1 concentrations do not assist in differentiating exudative effusions. PMID- 12911826 TI - Elevated levels of circulating adhesion molecules in patients with active pulmonary tuberculosis. AB - OBJECTIVE: Recent studies have indicated the importance of cell adhesion molecules in the pathogenesis of various inflammatory lung diseases. Our study was designed to determine whether five soluble adhesion molecules including soluble L-, E- and P-selectin (sL-, sE- and sP-selectin), intercellular adhesion molecule-1 (sICAM-1), and vascular cell adhesion molecule-1 (sVCAM-1) in serum reflect the severity of active pulmonary tuberculosis (TB), and whether there is a distinct profile of these soluble molecules in this disease. METHODOLOGY: Using enzyme-linked immunosorbent assays, we measured the serum levels of these five soluble adhesion molecules in 31 patients with active TB and 11 healthy volunteers. RESULTS: Serum levels of sE-selectin, sP-selectin and sICAM-1, but not sL-selectin or sVCAM-1, were significantly higher in patients with active TB than in the control subjects (P < 0.001, each). Significant correlations were detected only between serum levels of sE-selectin and sP-selectin, sE-selectin and sICAM-1, and sP-selectin and sICAM-1. There was a significant correlation between the Gaffky scale result (a scale assessing the number of mycobacteria bacilli present) and all of the above adhesion molecules, except for sL-selectin. Serum levels of sE-selectin, sL-selectin and sICAM-1 also correlated with the CXR radiological score. Higher levels of sL-selectin and sICAM-1 were detected in the serum of patients with radiological cavity formation compared to those without. The ESR, C-reactive protein and circulating neutrophil counts all correlated significantly with sE-selectin, sP-selectin, sICAM-1 and sVCAM-1. CONCLUSION: The results suggest that there is a distinct profile of soluble adhesion molecules in active pulmonary TB and that sE-selectin, sP-selectin, and especially sICAM-1 appear to be the most sensitive clinical measures of disease severity. PMID- 12911827 TI - Factors associated with improvement in breathing capacity during exercise in patients with chronic obstructive pulmonary disease. AB - OBJECTIVES: The aim of this study was to explore the relationship between resting pulmonary function indices and the ratio of minute ventilation at peak exercise to the maximal voluntary ventilation (VEmax/MVV) and to determine whether an improvement in breathing capacity during exercise (i.e. VEmax/MVV > 1) is associated with greater exercise capacity in patients with COPD. METHODOLOGY: The results of pulmonary function tests and incremental, symptom-limited cardiopulmonary exercise testing in 84 patients with predominantly moderate to severe COPD were reviewed. Multiple linear regression analysis was applied to determine the relationship of VEmax/MVV with selected independent variables at rest. Multiple logistic regression was used to determine significant predictors of VEmax/MVV 1. RESULTS: FEV1/FVC and inspiratory capacity (IC) were the only variables among resting pulmonary function indices that were significant independent determinants of VEmax/MVV and the stepwise analysis generated the following equation: VEmax/MVV = (-1.05E-02 x FEV1/FVC) + (0.15 x IC) + 1.28; r= 0.701, P < 0.001. Using multiple logistic regression with VEmax/MVV 1 as a dependent categorical variable, FEV1/FVC was the only significant predictor among resting pulmonary indices of a VEmax/MVV ratio of > 1 (Odds ratio 0.93, 95%CI 0.89, 0.97). There was a significant association between VEmax/MVV and peak oxygen uptake (VO2max) after adjusting for FEV1 (r = 0.66, P < 0.001). If the categorical variable of VEmax/MVV ( 1) was used instead of a continuous variable, a significant association with VO2max remained after adjusting for FEV1 (r = 0.60, P < 0.001). CONCLUSIONS: Among resting pulmonary function indices, the FEV1/FVC ratio is the best determinant of an improvement in breathing capacity during exercise in COPD patients. After adjusting for FEV1, an improvement in breathing capacity during exercise is associated with significantly higher exercise capacity. PMID- 12911828 TI - Clustering effects of chronic obstructive pulmonary disease specific quality of life in hospitalized patients. AB - OBJECTIVES: The primary aim of treatments for COPD is to improve health-related quality of life. However, little is known of the clustering effects related to health-related quality of life as an outcome measure. If clustering effects are observed, these have important implications for sample size estimates when cluster randomization is used in interventional studies. This study aimed to determine the intracluster correlation coefficient (ICC) of the quality of life, between hospitals for COPD patients. METHODOLOGY: The Dyspnoea Impact and Symptoms Questionnaire was administered to 100 COPD inpatients from four public hospitals (25 from each) around metropolitan Melbourne, selected on the basis that they had not had any major programs implemented within the last 2 years that aimed to improve the management of COPD. Data were collected concerning demographic and socioeconomic variables and comorbidities. RESULTS: The highest ICC value for a health-related quality of life subscale was 0.02 (psychological score), while the highest for a symptom-based subscale was 0.04. CONCLUSIONS: There is minimal clustering effect of quality of life in COPD patients between the hospitals studied. Despite this, when using a cluster randomised design the sample size needed to detect the same effect as a study using simple randomisation could be inflated by up to 183%. If cluster randomization is required, the average cluster size should be kept as small as possible to negate this effect. PMID- 12911829 TI - Clinical and CT characteristics of Chinese medicine-induced acute respiratory distress syndrome. AB - OBJECTIVE: The aim of the present study was to explore the clinical and radiographic characteristics of Chinese medicine-induced acute respiratory distress syndrome (ARDS), focusing on high resolution CT (HRCT) findings. METHODOLOGY: Five patients diagnosed with Chinese medicine-induced ARDS were enrolled in the study. Details regarding clinical history, previous medical history, and laboratory data were collected and reviewed. The extent of involvement of each HRCT abnormality was assessed for each of three zones of each lung and was estimated to the nearest 10% of parenchymal involvement. RESULTS: The causative Chinese medicines were as follows: Sho-saiko-to (two cases), Sai rei-to (two cases) and Otsu-ji-to (one case). Scutellariae radix was the common ingredient in these drugs. The mean duration of medication use before the onset of ARDS was 22 days (range 7-49 days). High levels of C-reactive protein and lactate dehydrogenase (LDH), and mild liver dysfunction were common laboratory findings. The major HRCT findings were as follows: ground-glass opacity 50.2% (range 37-68%); airspace consolidation 16.0% (7-27%); spared area 26.6% (7-45%). In addition, the abnormalities tended to predominate in the peribronchovascular regions. Furthermore, architectural distortion was seen in all five cases, interlobular septal thickening occurred in four, irregular interface in four, and interlobar effusion in two. Corticosteroid therapy was very effective, and partial pressure of arterial oxygen to fraction of inspired oxygen (PaO2/FIO2) improved in proportion to the decrease in LDH level. CONCLUSIONS: Patients with Chinese medicine-induced acute lung injury exhibited ARDS. However, their prognosis was satisfactory, which suggests that a hypersensitivity reaction or a permeability oedema with minimal epithelial injury may be involved in the pathophysiology. PMID- 12911830 TI - Significance of lung shrinkage on CXR as a prognostic factor in patients with idiopathic pulmonary fibrosis. AB - OBJECTIVE: The aim of this study was to determine whether the presence of lung shrinkage on CXR can predict diminished survival in patients with idiopathic pulmonary fibrosis (IPF)/usual interstitial pneumonia (UIP). METHODOLOGY: In a hospital-based cohort study 68 subjects diagnosed with IPF/UIP by surgical lung biopsy or at autopsy were observed for a mean of 7.6 years. The radiographic scores from Cherniack's method, pulmonary function tests, arterial blood gas, and haematological data were obtained at initial presentation. Longitudinal radiographic changes over a mean interval of 2.7 years were measured. Survival analysis was performed using Kaplan-Meier and Cox's proportional hazards regression analysis. RESULTS: At some point during the observation period 36 (53%) of 68 patients did not exhibit lung shrinkage and 32 (47%) of 68 patients showed lung shrinkage. Patients with lung shrinkage were more likely to have a diminished survival than those with lung preservation; median survival was 4.4 vs 7.8 years, respectively. Lung shrinkage during the observation period (hazard ratio, 3.89; 95% CI = 1.68-9.01; P= 0.001) was associated with lower rates of survival. CONCLUSION: In patients with IPF/UIP, lung shrinkage on CXR during the observation period was a poor prognostic factor. PMID- 12911831 TI - Antibiotic desensitization in adults with cystic fibrosis. AB - OBJECTIVE: Allergic reactions to antibiotics occur in up to 30% of patients with cystic fibrosis (CF). Repeated antibiotic exposure and immune hyper responsiveness increase the risk of allergic reactions and may limit antibiotic choice. Desensitization may allow the successful administration of an antibiotic despite previous allergy. We aimed to determine the success of antibiotic desensitization in patients with CF in an adult CF unit over a 7-year period. METHODOLOGY: A retrospective medical record review was performed on the 19 patients who had undergone antibiotic desensitization procedures. Data collected included drug allergy and intolerance profiles, nature of allergies, and the outcome of desensitization procedures. Desensitization procedures were performed in a ward setting according to published methods. RESULTS: Nineteen patients (13 females) reported 62 drug allergies with a mean of 3.3 per patient. Of the 71 desensitization procedures undergone by this group, 54 (76%) were successful. Fifteen of the 19 patients were allergic to two or more beta-lactam antibiotics. Over half of the patients were desensitized to more than one antibiotic. Nine different antibiotics were used in 31 different patient/drug combinations. A successful outcome was achieved in 18/31 (58%) combinations, with three requiring treatment for mild allergic reactions. Allergic reactions caused drug cessation in a total of 19 patient/drug combinations (three after initial successful desensitization and full courses of antibiotics). Over 50% of these reactions occurred on day 1. Desensitization failures were more common in patients with well-documented allergic reactions to a specific drug. CONCLUSION: This study demonstrates that multiple antibiotic allergies are common in adults with CF. Cross-reactivity between beta-lactam antibiotics may limit antibiotic choice for the treatment of pulmonary exacerbations. Antibiotic desensitization allows safe and successful treatment in the ward setting of many patients with previous allergies to an antibiotic. In many patients symptoms of allergy still occur and result in cessation of the antibiotics. Use of corticosteroids and antihistamines may improve the success rate of desensitization procedures. PMID- 12911832 TI - A novel approach for quality control of total lung capacity in the clinical pulmonary function laboratory: a study in a veteran population. AB - OBJECTIVE: Quality control in the clinical pulmonary function laboratory has been well developed for spirometry and diffusing capacity but not for the measurement of TLC. The purpose of the present study was to test two approaches to this problem. First, we compared TLC by body plethysmography (TLCb) with a value predicted from TLC measured by multibreath helium dilution (TLCm). Concordance between the measured and predicted values would imply the validity of the measurements. Second, we measured the test-retest variability of TLCb, TLCm and TLC measured by single breath helium dilution (TLCs) to assess the consistency of the measurements. METHODOLOGY: We performed a prospective study of 815 veterans. RESULTS: The prediction of TLCb from TLCm improved (r2 increased from 0.44 to 0.64) when FEV1/FVC and the difference between TLCm and TLCs were added to the model. The coefficient of variation for test-retest of TLCs, TLCm and TLCb were 8.9, 7.1 and 5.4%, respectively. Of all tests, 5.9% were inconsistent based on pathophysiology or measurement error and attributed mostly to TLCm. CONCLUSIONS: Prediction of TLCb from TLCm was not sufficiently accurate as to be useful for quality control. Comparison of TLCs, TLCm and TLCb may be useful for determining the internal data validity. PMID- 12911833 TI - Improvement in health-related quality of life with fluticasone propionate compared with budesonide or beclomethasone dipropionate in adults with severe asthma. AB - OBJECTIVE: Changes in health-related quality of life (HRQoL) were evaluated in adults with severe asthma following inhaled corticosteroid treatment with high dose beclomethasone dipropionate or budesonide (BDP/BUD) and compared with fluticasone propionate taken at approximately half the dose of BDP/BUD. METHODOLOGY: HRQoL was assessed as part of an open, multicentre, randomized, parallel-group study in Australia evaluating the safety and efficacy of switching to fluticasone propionate (FP) 1000-2000 micro g/day (n = 67) compared with remaining on BDP/BUD >/=1750 micro g/day (n = 66) for 6 months. Patients completed two HRQoL questionnaires, the Asthma Quality of Life Questionnaire (AQLQ) and the Medical Outcomes Study Short Form-36 (SF-36), at baseline and at weeks 12 and 24. A change in AQLQ score of >/=0.5 was considered to be clinically meaningful. RESULTS: There were significant improvements in HRQoL with FP on four of the eight dimensions on the SF-36 (i.e. physical functioning, general health, role-emotional, and mental health), while there were no significant improvements in HRQoL in the BDP/BUD group. Overall, patients in the FP group experienced significantly greater improvement (P < 0.001) in AQLQ scores at weeks 12 and 24 compared with the BDP/BUD group. On the individual domains of the AQLQ, there were significant treatment differences (P < 0.01) in favour of FP in three of the four domains (activity limitations [0.92], symptoms [0.73], and emotional function [1.02]). Mean differences between groups for overall score and these three domains were also clinically meaningful. CONCLUSION: Patients with severe asthma who received FP (at approximately half the dose of BDP/BUD) experienced statistically significant, as well as clinically meaningful, improvements in their HRQoL. PMID- 12911834 TI - Control of an outbreak of BCG complications in Gaza. AB - OBJECTIVE: We aim to describe Bacille Calmette Guerin (BCG) complications in Gaza using two studies: one during an outbreak and the other after control of the possible contributing factors to this outbreak. METHODOLOGY: The first study was conducted on 6145 newborn infants vaccinated in 21 primary care centres in Gaza with BCG vaccine, Pasteur Paris, batch number 5122 from July to October 2001. The study was repeated after changing the BCG vaccine and training 63 nurses from November to December 2001. The training program included theoretical lectures on BCG and tuberculosis and practical training in strict intradermal injection. The second study included 6877 newborn infants vaccinated with BCG vaccine, batch number 101023, Denmark, from January to April 2002. RESULTS: During the outbreak, BCG complications occurred in 225 infants with a complication rate of 36.61 per 1000 vaccinations. The mean age at presentation was 4 months. The commonest complications were regional lymphadenitis in 138 (61.33%) infants, local abscess in 48 (21.33%) infants, local ulcer in 26 (11.56%) infants, keloid scar in 12 (5.33%) infants and one (0.44%) infant who died from disseminated disease had severe combined immunodeficiency. In the follow-up study, BCG complications occurred in 43 infants with a complication rate of 6.25 per 1000 vaccinations, which is significantly lower (P < 0.001) than that during the initial outbreak. CONCLUSION: The study supports the use of a less virulent vaccine and proper vaccination techniques to minimize the incidence of BCG complications. The training of nurses in strict intradermal injection should be maintained and the proper selection of those receiving the vaccines should be considered to avoid the vaccination of any infant with immunodeficiency. PMID- 12911835 TI - Comment on respiratory morbidity and lung function. PMID- 12911836 TI - Risks of serious complications and death from smallpox vaccination: a systematic review of the United States experience, 1963-1968. AB - BACKGROUND: The United States (US) has re-instituted smallpox vaccinations to prepare for an intentional release of the smallpox virus into the civilian population. In an outbreak, people of all ages will be vaccinated. To prepare for the impact of large-scale ring and mass vaccinations, we conducted a systematic review of the complication and mortality risks of smallpox vaccination. We summarized these risks for post-vaccinial encephalitis, vaccinia necrosum (progressive vaccinia), eczema vaccinatum, generalized vaccinia, and accidental infection (inadvertant autoinoculation). METHODS: Using a MEDLINE search strategy, we identified 348 articles, of which seven studies met our inclusion criteria (the number of primary vaccinations and re-vaccinations were reported, sufficient data were provided to calculate complication or case-fatality risks, and comparable case definitions were used). For each complication, we estimated of the complication, death, and case-fatality risks. RESULTS: The life threatening complications of post-vaccinial encephalitis and vaccinia necrosum were at least 3 and 1 per million primary vaccinations, respectively. Twenty-nine percent of vaccinees with post-vaccinial encephalitis died and 15% with vaccinia necrosum died. There were no deaths among vaccinees that developed eczema vaccinatum; however, 2.3% of non-vaccinated contacts with eczema vaccinatum died. Among re-vaccinees, the risk of post-vaccinial encephalitis was reduced 26-fold, the risk of generalized vaccinia was reduced 29-fold, and the risk of eczema vaccinatum was reduced 12-fold. However, the risk reductions of accidental infection and vaccinia necrosum were modest (3.8 and 1.5 fold respectively). PMID- 12911837 TI - Founder mutations in BRCA1/2 are not frequent in Canadian Ashkenazi Jewish men with prostate cancer. AB - BACKGROUND: Relatives of BRCA1 and BRCA2 mutation carriers have long been proposed by epidemiological studies to have an increased risk of developing prostate cancer. In the Ashkenazi Jewish (AJ) population, the existence of 3 frequent founder mutations, 185delAG and 5382insC in BRCA1 and 6174delT in BRCA2 greatly facilitates screening for carriers. METHODS: We tested 146 AJ men with confirmed diagnoses of invasive prostate cancer. Thirteen had at least one first degree relative with prostate cancer. The median age at diagnosis of participants was 67.9 years (range 48.6-84.2 years). Subjects were screened for the BRCA1:185delAG, BRCA1:5382insC and BRCA2:6174delT mutations simultaneously using a multiplex sizing assay detecting band shifts in the presence of the variant sequence. RESULTS: Two out of 146 individuals were found to carry the germline BRCA2 mutation 6174delT (1.4%); the previously reported population frequency for this mutation is ~1% in AJ. We found no BRCA1 mutations. One carrier had 2 uncles affected with prostate cancer, while the other had an uncle and daughter with breast cancer. We combined our results with previously published data examining these 3 founder AJ mutations in men with prostate cancer and in population controls. Including our results, studies to date reported 5/463 (1.1%), 2/293 (0.68%) and 7/461 (1.3%) carriers for the BRCA1:185delAG, BRCA1:5382insC and BRCA2:6174delT mutations in prostate cancer cases, respectively. This compares with combined reported frequencies of 85/9371 (0.91%), 24/8867 (0.27%) and 119/9514 (1.3%) for the same mutations in control individuals. There was no statistically significant excess of mutations in cases compared to controls in either gene. CONCLUSIONS: Our observations remain preliminary. By combining all studies published to date, we have an 80% power to detect ORs of 2.7, 6.6 and 2.5 (185delAG, 5382insC and 6174 delT, respectively) while the values we observed range between 1.0 and 2.5. However, the contribution of rare mutations with such low odds ratios to the population prostate cancer burden is unlikely to be large enough to be clinically useful. Thus, contrary to suggestions from some previous epidemiological data, our observations do not support an important role for AJ founder BRCA1/2 mutations in prostate cancer risk. PMID- 12911838 TI - Dapsone induced cholangitis as a part of dapsone syndrome: a case report. AB - BACKGROUND: Dapsone can rarely cause a hypersensitivity reaction called dapsone syndrome, consisting of fever, hepatitis, exfoliative dermatitis, lymphadenopathy and hemolytic anemia. Dapsone syndrome is a manifestation of the DRESS (drug rash with eosinophilia and systemic symptoms) syndrome which is a serious condition that has been reported in association with various drugs. Cholangitis in dapsone syndrome has not been reported so far in the world literature. CASE PRESENTATION: We report a patient who presented with fever, exfoliative dermatitis, jaundice and anemia within three weeks of starting of dapsone therapy. These features are typical of dapsone syndrome, which is due to dapsone hypersensitivity and is potentially fatal. Unlike previous reports of hepatitic or cholestatic injury in dapsone syndrome we report here a case that had cholangitic liver injury. It responded to corticosteroids. CONCLUSION: We conclude that cholangitis, though unusual, can also form a part of dapsone syndrome. Physicians should be aware of this unusual picture of potentially fatal dapsone syndrome. PMID- 12911839 TI - Analysis of antimicrobial susceptibility and virulence factors in Helicobacter pylori clinical isolates. AB - BACKGROUND: In this study, we evaluated the prevalence of primary resistance of Brazilian H. pylori isolates to metronidazole, clarithromycin, amoxicillin, tetracycline, and furazolidone. In addition, the vacA, iceA, cagA and cagE genotypes of strains isolated from Brazilian patients were determined and associated with clinical data in an effort to correlate these four virulence markers and antibiotic resistance. METHODS: H. pylori was cultured in 155 H. pylori-positive patients and MICs for metronidazole, clarithromycin, amoxicillin, tetracycline, and furazolidone were determined by the agar dilution method. Genomic DNA was extracted, and allelic variants of vacA, iceA, cagA and cagE were identified by the polymerase chain reaction. RESULTS: There was a strong association between the vacA s1/cagA -positive genotype and peptic ulcer disease (OR = 5.42, 95% CI 2.6-11.3, p = 0.0006). Additionally, infection by more virulent strains may protect against GERD, since logistic regression showed a negative association between the more virulent strain, vacA s1/cagA-positive genotype and GERD (OR = 0.26, 95% CI 0.08-0.8, p = 0.03). Resistance to metronidazole was detected in 75 patients (55%), to amoxicillin in 54 individuals (38%), to clarithromycin in 23 patients (16%), to tetracycline in 13 patients (9%), and to furazolidone in 19 individuals (13%). No significant correlation between pathogenicity and resistance or susceptibility was detected when MIC values for each antibiotic were compared with different vacA, iceA, cagA and cagE genotypes. CONCLUSION: The analysis of virulence genes revealed a specific association between H. pylori strains and clinical outcome, furthermore, no significant association was detected among pathogenicity and resistance or susceptibility. PMID- 12911840 TI - Effects of disruption of heat shock genes on susceptibility of Escherichia coli to fluoroquinolones. AB - BACKGROUND: It is well known that expression of certain bacterial genes responds rapidly to such stimuli as exposure to toxic chemicals and physical agents. It is generally believed that the proteins encoded in these genes are important for successful survival of the organism under the hostile conditions. Analogously, the proteins induced in bacterial cells exposed to antibiotics are believed to affect the organisms' susceptibility to these agents. RESULTS: We demonstrated that Escherichia coli cells exposed to levofloxacin (LVFX), a fluoroquinolone (FQ), induce the syntheses of heat shock proteins and RecA. To examine whether the heat shock proteins affect the bactericidal action of FQs, we constructed E. coli strains with mutations in various heat shock genes and tested their susceptibility to FQs. Mutations in dnaK, groEL, and lon increased this susceptibility; the lon mutant exhibited the greatest effects. The increased susceptibility of the lon mutant was corroborated by experiments in which the gene encoding the cell division inhibitor, SulA, was subsequently disrupted. SulA is induced by the SOS response and degraded by the Lon protease. The findings suggest that the hypersusceptibility of the lon mutant to FQs could be due to abnormally high levels of SulA protein resulting from the depletion of Lon and the continuous induction of the SOS response in the presence of FQs. CONCLUSION: The present results show that the bactericidal action of FQs is moderately affected by the DnaK and GroEL chaperones and strongly affected by the Lon protease. FQs have contributed successfully to the treatment of various bacterial infections, but their widespread use and often misuse, coupled with emerging resistance, have gradually compromised their utility. Our results suggest that agents capable of inhibiting the Lon protease have potential for combination therapy with FQs. PMID- 12911841 TI - Peroxisome proliferator activated receptors and stroke. PMID- 12911842 TI - Losartan reduces cardiovascular morbidity and mortality in patients with hypertension and left ventricular hypertrophy. PMID- 12911843 TI - C-reactive protein and risk of cardiovascular disease: evidence and clinical application. PMID- 12911844 TI - Microalbuminuria and diabetic cardiovascular disease. PMID- 12911845 TI - Implications of the heart protection study for reducing coronary events in high risk patients. PMID- 12911846 TI - The insulin resistance syndrome. PMID- 12911847 TI - Isoprenoid metabolism and the pleiotropic effects of statins. PMID- 12911848 TI - Evidence-based management of dyslipidemias in women. PMID- 12911849 TI - Reverse cholesterol transport: high-density lipoprotein's magnificent mile. PMID- 12911850 TI - In-hospital initiation of statin therapy in patients with acute coronary events. PMID- 12911851 TI - Toxicity of antilipidemic agents: facts and fictions. PMID- 12911852 TI - Magnetic resonance imaging of coronary atherosclerosis. PMID- 12911853 TI - Niacin: a powerful adjunct to other lipid-lowering drugs in reducing plaque progression and acute coronary events. PMID- 12911854 TI - Health hazards for terrestrial vertebrates from toxic cyanobacteria in surface water ecosystems. AB - Toxigenic cyanobacteria are photosynthetic prokaryotes that are most often recognized in marine and freshwater systems, such as lakes, ponds, rivers, and estuaries. When environmental conditions (such as light, nutrients, water column stability, etc.) are suitable for their growth, cyanobacteria may proliferate and form toxic blooms in the upper, sunlit layers. The biology and ecology of cyanobacteria have been extensively studied throughout the world during the last two decades, but we still know little about the factors and processes involved in regulating toxin production for many cyanobacterial species. In this minireview, we discuss these microorganisms, and more especially the toxins they produce, as a potential and important health risk for wild and domestic animals. PMID- 12911855 TI - Renal handling of calcium and phosphorus in experimental renal hyperparathyroidism in dogs. AB - Twenty-four hour urinary excretion, fractional excretion and the filtered load of calcium and phosphorus were monitored as hyperparathyroidism evolved in a model of progressive canine renal failure. Thirteen beagles of both sexes aged four and a half months were used. Nine of them were subjected to a renal damaging schedule (neomycine, 60 mg/kg/48 h, IM, 32 weeks) in order to induce chronic renal failure leading to secondary hyperparathyroidism (2HPT group). The remaining four were kept as the control group. The experiment was conducted over 32 weeks. Blood and 24 h urine were collected every four weeks. Calcium, phosphorus and creatinine were analyzed. Plasma parathormone and calcitonin were determined at weeks 0, 12, 24 and 32. The level of renal function in the 2HPT animals was reduced to 25% of that of the controls (endogenous creatinine clearance was 0.45 +/- 0.22 mL/min/kg as opposed to 1.81 +/- 0.54 mL/min/kg). Hyperparathyroidism was confirmed by a progressive increase in the levels of the parathyroid hormone. Calcitonin levels were not modified. A tendency to hypocalcaemia was observed, reaching statistically significant levels from the twenty-eighth week of the study, when hyperphosphataemia also became significant. Daily urinary excretion of calcium and phosphorus remained at values considered normal throughout the experiment with no alteration imputable to the impaired renal function. This is explained by the decrease in the filtered load of these elements (in both cases statistically significant from the 24th week on) being associated with an increase in their fractional excretion. Thus, calcium and phosphorus urinary excretion values could be maintained in a normal range up to the end of the experiment, showing that renal calcium handling in dogs with experimentally induced renal failure seems to differ from that observed in human patients. PMID- 12911856 TI - Humoral and cell-mediated immune responses of d/d histocompatible pigs against classical swine fever (CSF) virus. AB - A better understanding of cell-mediated immune responses to classical swine fever virus (CSFV) is essential for the future development of improved vaccines. We analyzed the generation of cell-mediated and humoral immune responses in d/d histocompatible pigs following CSFV infection or vaccination. Viral infection induced high T cell responses with high primary and secondary CTL activity correlated with high IFN-gamma production, whereas vaccination with a live vaccine followed by infection mainly induced neutralizing antibody but low cell mediated responses. Moreover, high IgG1 response was associated with high IFN gamma response following infection whereas a weak IFN-gamma response was related to a good IgG2 response but a low IgG1 production. These data could reflect Th1/Th2-like balance of immune responses depending upon immunization protocols, which has not yet been described in the pig. T-cell responses to CSFV were evidenced by CSFV-specific CD25 upregulation on CD4-CD8+, but not on CD4+CD8- cells, which further illustrated the importance of CTL responses after infection. Our results indicated that generation of cell-mediated immune responses was much higher following intranasal/oral CSFV infection than after intramuscular vaccination, which implies that the capacity of new CSFV vaccines to induce higher T-cell responses should be considered. PMID- 12911857 TI - In vitro formation of metabolic-intermediate cytochrome P450 complexes in rabbit liver microsomes by tiamulin and various macrolides. AB - Tiamulin and a number of macrolides were evaluated as to their ability in forming metabolic-intermediate (MI) complexes with cytochrome P450 in liver microsomes from rabbits bred for meat production. Complex formation, which occurred only in preparations where the expression of P450 3A was increased as the result of rifampicin pre-treatment and with different kinetics, was in the order tiamulin > erythromycin > TAO approximately roxithromycin approximately tylosin and did not take place with tilmicosin and spiramycin. Most of the tested compounds underwent an oxidative N-dealkylation and a good relationship could be found between the rate of N-dealkylase activity in induced preparations and the aptitude in generating MI complexes. Although the results from in vitro studies should be interpreted with caution, it is suggested that the potential for in vivo drug interactions also exists in the rabbit for tiamulin and for four out of the six tested macrolides. PMID- 12911858 TI - Functional analysis of recombinant bovine CD14. AB - Studies in mice and humans indicate that membrane CD14 (mCD14) on the surface of monocytes, macrophages and polymorphonuclear neutrophils (PMN) mediate activation of these cells by lipopolysaccharide (LPS). Soluble CD14 (sCD14), in the circulation, binds to LPS and blocks LPS binding to mCD14. The role of bovine CD14 in cellular activation by LPS is undefined. Changes in CD18 expression on PMN and steady state levels of mRNA for tumor necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6) and IL-8, sensitive markers for activation of leukocytes by LPS, were used to measure functional activity of recombinant bovine sCD14 (rbosCD14). Whole blood (n=3 cows) treated with LPS alone caused CD18 expression on PMN to increase by 12% (P<0.02), whereas pre-incubation of LPS with 10 or 100 microg/mL of rbosCD14 completely inhibited increase in CD18 expression. After treating whole blood with LPS at concentrations of 1, 100 or 10(4) ng/mL for 2 h, level of mRNA for TNF-alpha, IL-6 and IL-8 in leukocytes and concentration of TNF alpha in plasma increased. However, pre-incubation of LPS with rbosCD14 inhibited the increase in TNF-alpha mRNA, but not the increase in IL-6 and IL-8 mRNA. Excess amount of anti-human CD14 monoclonal antibody (MAB) also inhibited LPS induced increase in TNF-alpha mRNA. Preincubation of LPS with rbosCD14, or rbosCD14 plus MAB did not affect LPS-induced increase in TNF-alpha in plasma. Collectively, results indicate that rbosCD14 inhibit LPS-induced increase in CD18 expression and TNF-alpha mRNA. However, secretion of TNF-alpha was not inhibited by pre-incubation of LPS with rbosCD14. The TNF-alpha in plasma may partially induce transcription of IL-6 and IL-8, which contribute to the CD14-independent increase in level of mRNA for IL-6 and IL 8. PMID- 12911859 TI - Experimental Coxiella burnetii infection in pregnant goats: excretion routes. AB - Q fever is a widespread zoonosis caused by Coxiella burnetii. Infected animals, shedding bacteria by different routes, constitute contamination sources for humans and the environment. To study Coxiella excretion, pregnant goats were inoculated by the subcutaneous route in a site localized just in front of the shoulder at 90 days of gestation with 3 doses of bacteria (10(8), 10(6) or 10(4) i.d.). All the goats aborted whatever the dose used. Coxiella were found by PCR and immunofluorescence tests in all placentas and in several organs of at least one fetus per goat. At abortion, all the goats excreted bacteria in vaginal discharges up to 14 days and in milk samples up to 52 days. A few goats excreted Coxiella in their feces before abortion, and all goats, excreted bacteria in their feces after abortion. Antibody titers against Coxiella increased from 21 days post inoculation to the end of the experiment. For a Q fever diagnostic, detection by PCR and immunofluorescence tests of Coxiella in parturition products and vaginal secretions at abortion should be preferred to serological tests. PMID- 12911860 TI - Humoral response (IgG) of goats experimentally infected with Fasciola hepatica against cysteine proteinases of adult fluke. AB - The use of cysteine proteinases from Fasciola hepatica adult flukes for the serodiagnosis of caprine fasciolosis by means of an indirect ELISA test was studied. Two proteolytic fractions from adult fluke homogenates, with apparent molecular weights of 28 and 34 kDa (P28 and P34 respectively), were characterised as cysteine proteinases using azocasein assays and gelatin gel analysis. Both P28 and P34 fractions were electroluted and used as antigens in two different indirect ELISA tests. Serum IgG levels against P28 and P34 in goats given an experimental primary infection with 200 metacercariae or in goats given two experimental infections with 200 metacercariae were determined and compared with those observed in an uninfected control group. ELISA tests using both cysteine proteases showed a rapid and consistent detection of specific IgG in all experimentally infected goats. The IgG response to P28 was the first to be detected as early as 2-3 weeks post-infection and remained elevated throughout the experiment. The response to P34 was detected later (4-6 wpi) and disappeared in some animals at 18 wpi, while flukes were still present in the bile ducts. No significant differences were observed between the anti-P28 and anti-P34 IgG responses between animals receiving a primary or a challenge infection. The results of our study, although preliminary, are promising since the P28 ELISA described here may be a reliable method for the immunodiagnosis of F. hepatica infection in goats. PMID- 12911861 TI - A pharmacokinetic model to document the actual disposition of topical ivermectin in cattle. AB - Ivermectin is a worldwide-used antiparasitic drug largely administered to cattle as a topical formulation (pour-on). The actual plasma and faecal disposition of pour-on ivermectin in cattle was documented using an original pharmacokinetic model, and taking into account the oral ingestion of the topical drug following physiological licking as a secondary route of exposure. Six pairs of monozygotic twin cattle received successively one i.v. and two pour-on administrations of ivermectin at a 3-5-month interval. For one pour-on administration, the twins were separated into an unrestrained group and a group where self- and allo licking were prevented. Ivermectin concentrations in the plasma and faeces were determined by HPLC. Licking resulted in a high intra-and inter-individual variability of systemic exposure after topical application. By the means of pharmacokinetic modelling, we showed that 58-87% of the pour-on dose was ingested, while only 10% was absorbed percutaneously. Approximately 72% of the ingested ivermectin transited directly into the faeces, resulting in a 7-fold higher faecal excretion of the parent drug than in the non-lickers. We conclude that topical administration does not guarantee a controlled drug delivery in cattle. More importantly, the simulations revealed that non-treated cattle could get easily contaminated by allo-licking, raising the public health problem of unexpected drug residues in edible tissues. PMID- 12911862 TI - Multispecies and multiple anthelmintic resistance on cattle nematodes in a farm in Argentina: the beginning of high resistance? AB - There are very few resistance records on cattle nematodes. South American successive records have been increasing rapidly since the year 2000. In Argentina, increasing dissatisfaction based on the exclusive use of macrocyclic lactones has prompted the use of benzimidazoles in the Pampean region. The studied farm is located in the Argentina humid Pampas and had apparently poor results after anthelmintic treatments. Evaluation of resistance was firstly based on faecal egg reduction after treatment in November 2001 and June 2002 and was complemented with worm counts obtained by necropsy in June 2002. The study reports that the reduction of faecal egg excretion after benzimidazole (reduction 31-79%) or ivermectin treatments (76-97%) in November 2001 was insufficient and these poor results were confirmed in June 2002 (benzimidazoles (65-89%) and ivermectin (47-77%)). Several methods for evaluating faecal egg count reduction were performed and yielded different results. A new method based on the estimated faecal egg counts (corrected from the initial faecal egg counts and animal type using a general linear model) was used and gave higher flexibility in the interpretation of putative resistance to anthelmintics. From necropsy results, it could be concluded that Cooperia oncophora was resistant to avermectins and benzimidazoles and that Cooperia punctata, Ostertagia ostertagi and Haemonchus placei were resistant to benzimidazoles. This case of multispecies and multidrug resistance is probably not unique and could reflect the emergence of resistances in Argentina. This emergence is probably due to the intensive use of anthelmintics, the absence of refugia, and the frequent circulation of infected cattle. PMID- 12911863 TI - Evaluation of murine liver transmission electron micrographs by an innovative object-based quantitative image analysis system (Cellenger). AB - BACKGROUND: Transmission electron micrographs are widely used to demonstrate tissue damage. However, the results are qualitative and dependent on the experience of the investigator. Recently, a new multiscale object-based quantitative image analyzing systems (Cellenger) has been introduced to study highly textured black-and-white images. It is unknown, whether this system permits the quantitative image analysis of electron micrographs of parenchymal tissue. Therefore, we analyzed whether the Cellenge system permits the quantitative evaluation of electron micrographs of murine liver under normal conditions and after ischemia-reperfusion injury. The results were compared with those obtained by conventional qualitative classification. - METHOD: Transmission electron micrographs from murine liver that had been exposed to isolated reversible ischemia at hypothermic conditions of 4 degrees C, 15 degrees C, 26 degrees C and 37 degrees C, and of sham-operated animals, which served as controls (2 images per animal, n = 3 in each group), were analyzed qualitatively by an investigator with experience in electron microscopy. For quantitative analysis, the Cellenger was used and the following damage parameters were studied: ratio of area of endothelial cell nucleus to area of endothelial cell (N/C ratio), ratio of area of hepatocellular vacuoles to area of total hepatocyte cytoplasm (V/C ratio) and ratio of area of microvilli in the space of Disse to area of the sinusoids (M/S ratio). All values were sampled within one group (n=6) and the data given in [%] (MW +/- SEM). P-values were accepted as significant below 0.05. RESULTS: After normothermic ischemia, all quantitative damage parameter were significantly altered as compared to sham-operated animals (N/C 15 +/- 9% vs. 37 +/- 7%, V/C 18 +/- 4% vs. 0, and M/S 0 vs. 10 +/- 1%) and all hypothermia groups. The qualitative electron micrograph section analysis corresponded very well with these results. CONCLUSION: We demonstrate that an multiscale object-based quantitative analysis of transmission electron micrographs from mouse liver under control conditions and after I/R provide accurate classification of relevant tissue damage parameter. The system is now ready to use for further applications within the field of highly textured electron micrographs. PMID- 12911864 TI - Effect of smoking on risk factors for cardiovascular disease in patients with diabetes mellitus and renal insufficiency. AB - BACKGROUND: Many studies have been published demonstrating a strong correlation between smoking, renal lesions and cardiovascular morbidity and mortality. Possible contributing factors are elevated blood pressure values, changes in vascular reactivity, concentrations of lipids, fibrinogen and carboxyhemoglobin, in patients with diabetes mellitus, the quality of diabetes control and insulin resistance. Other possible risk factors may include advanced glycation end (AGE) products, total plasma homocysteine and metabolites as well as symmetrical (SDMA) and asymmetrical (ADMA) dimethylarginine. It was the goal of the present trial to investigate the serum concentrations of these >new<, possible risk factors in a selection free population of patients with type 1 and insulin-treated type 2 diabetes mellitus, in patients with chronic renal insufficiency and in renal transplant recipients. The second aim was to analyse the effect of cigarette smoking on the levels of these laboratory parameters. PATIENTS AND METHODS: A total of 544 subjects (n = 98 patients with chronic renal insufficiency without hemodialysis, n = 84 patients with hemodialysis, n = 50 renal transplant recipients, n = 114 patients with type 1 and n = 147 patients with insulin treated type 2 diabetes mellitus, n = 51 healthy controls) participated in the trial. RESULTS: Patients with renal insufficiency had significantly higher concentrations of the AGE-products N-epsilon-Carboxymethyllysine (CML) and pentosidine compared with both groups of patients with insulin-treated diabetes mellitus and the healthy controls. In patients with type 1 diabetes there was a positive correlation between HbA1c and the concentration of CML (r = 0.405, p = 0.017) and between the pentosidine-concentration and the serum creatinine-level (r = 0.482, p = 0.001). In type 2 diabetes there was a correlation between CML and diastolic blood pressure (r = 0.239, p = 0.039). In comparison to the healthy controls, patients with renal diseases and patients after kidney transplantation had higher concentrations of total plasma homocysteine and its metabolites. Similar results were found for SDMA and ADMA: Patients with chronic renal insufficiency both with and without hemodialysis had significantly higher values than the renal transplant recipients and healthy controls. A clear influence of cigarette smoking on the levels of the laboratory parameters measured could not be determined in any group; neither in the patients with diabetes mellitus, nor in the patients with renal diseases, nor in the healthy controls. CONCLUSIONS: The present trial demonstrates an increase in the parameters linked to the development of cardiovascular diseases including total plasma homocysteine, its metabolites, the dimethylarginines SDMA and ADMA and advanced glycation end products depending on the degree of renal insufficiency. Moreover, in patients with insulin-treated diabetes mellitus, the concentrations of the AGE-products CML and pentosidine seem to be strongly influenced by the quality of diabetes control and blood pressure levels. There was no influence of cigarette smoking on the levels of the laboratory parameters measured. PMID- 12911865 TI - Computed tomographic findings in bourneville-pringle disease. AB - Bourneville-Pringle disease (synonym: tuberous sclerosis, epiploia) is a neuroectodermal disorder. A characteristic clinical triad of adenoma sebaceum, mental retardartion, and seizures leads to early childhood diagnosis. Many patients show morphological changes of the visceral organs, vascular, and skeletal system. We review the computed tomographic findings of Bourneville Pringle disease as relates to an oligosymptomatic 36-year-old woman with late onset of clinical symptoms. PMID- 12911866 TI - Intrapulmonary administration of natural honey solution, hyperosmolar dextrose or hypoosmolar distill water to normal individuals and to patients with type-2 diabetes mellitus or hypertension: their effects on blood glucose level, plasma insulin and C-peptide, blood pressure and peaked expiratory flow rate. AB - Safety and effect intrapulmonary administration (by inhalation) of 60 % honey solution, 10% dextrose or distill water on blood sugar, plasma insulin and C peptide, blood pressure, heart rate, and peaked expiratory flow rate (PEFR) in normal or diabetic subjects were studied. - Twenty-four healthy subjects, 16 patients with type 11 diabetes mellitus and six patients with hypertension were entered for study. They were underwent complete physical examination and laboratory investigations. Twelve healthy subjects were subjected for distill water inhalation for 10 min, and after one week they received inhalation of honey solution (60% wt/v) for 10 min. Another 12 healthy subjects received inhalation of 10% dextrose for 10 min. Blood glucose level, plasma insulin and C-peptide, blood pressure, heart rate and PEFR were estimated before inhalation and during 2 3 hrs after inhalation, at 30 min intervals. Random blood glucose level was estimated in eight patients with poorly controlled diabetes mellitus, and repeated 30 min after honey inhalation. One week later, fasting blood glucose level was estimated in each patient and blood glucose level was re-estimated during three hrs after honey inhalation, at 30 min intervals. Glucose tolerance test was performed in another eight patients with type-2 diabetes mellitus, and after one week the procedure was repeated with inhalation of honey, which was started immediately after ingestion of glucose. Six hypertensive patients received honey inhalation for 10 min; supine blood pressure and heart rate were measured before and after inhalation. - Results showed that in normal subjects distill water caused mild elevation of blood glucose level, mild lowering of plasma insulin, and significant reduction of plasma C-peptide. 10% dextrose inhalation caused mild reduction of plasma insulin and C-peptide and unremarkable changes in blood glucose level. No significant changes were obtained in blood pressure, heart rate or PEFR after distill water or 10% dextrose inhalation. Honey inhalation caused lowering of blood glucose level and elevation of plasma insulin and C-peptide, mild reduction of blood pressure and up to 11 and 16 percent increase in PEFR. Honey inhalation significantly reduced random blood glucose level from 199 +/- 40.9 mg/dl to 156 +/- 52.3 mg/dl after 30 min (p = 0.0303). Fasting blood glucose level was reduced after honey inhalation during three hr post-inhalation, which was significant at hr three (p<0.05). Intensity of hyperglycemia was significantly lowered in glucose tolerance test when patients received honey inhalation. Systolic and diastolic blood pressure was reduced by honey inhalation in hypertensive patients; significant changes were obtained at 60 and 120 min after inhalation. No adverse effects were observed with inhalation of distill water, 10% dextrose and 60% honey solution except for nasal watery discharge experienced by all subjects and mild cough that was experienced by seven subjects after honey inhalation. - The results demonstrated that honey inhalation was safe and effective in reducing blood glucose level, in normal and diabetic subjects, it could improve glucose tolerance test, elevate plasma insulin and C-peptide and PEFR, and reduce elevated blood pressure in hypertensive patients. PMID- 12911867 TI - The importance of serial measurements of cytokine levels for the evaluation of their role in pathogenesis in familial Mediterraean fever. AB - Familial Mediterranean fever (FMF) is an autosomal recessive disorder characterized by recurrent fever of unknown origin, renal amyloidosis, peritonitis, pleuritis and/or synovitis. There have been many studies to elucidate the etiopathogenesis of FMF. IL-6 is a cytokine that can induce the formation of serum amyloid A and C-reactive protein, both of which are important in development of amyloidosis. IL-6 was determined to be strongly associated in the etiopathogenesis of periodic fever in Chinese-pei dogs. The dogs with this syndrome experience periodic fever, arthritis, renal amyloidosis, a clinical picture very alike of human FMF. Here, we aimed to study mainly whether IL-6 had a similar etiopathogenetic role in human FMF as in Chinese-pei dogs syndrome. The median IL-6 blood levels were found to be higher in patients with acute (n=8) FMF attack (1.85 U/ml) compared to those (n=33) with asymptomatic ones (1.0 U/ml) (p=0.16). There are mainly two results: first; the study should be designed with a larger sample size of patients with acute attack in order to alleviate underestimation of significance, second; sampling time may give various results because of dynamic changes of cytokine levels during acute attack period. PMID- 12911868 TI - Oxidative protein damage parameters in plasma in chronic experimental diabetes in rats. AB - An increase in oxidative stress may contribute to the development of oxidative protein damage (OPD) in the streptozotocin-diabetic rat. To show the effect of hyperglycemia in promoting OPD, we determined protein carbonyl (PCO), nitrotyrosine (NT), total thiol (T-SH) and advanced oxidation protein product (AOPP) levels as markers of OPD, and lipid hydroperoxide (LHP) levels as a marker of lipid peroxidation in plasma of acute and chronic diabetic male Sprague-Dawley rats and their controls. The levels of the studied markers, except NT, were determined by colorimetric methods. NT levels were measured by ELISA. Plasma PCO and AOPP levels of chronic diabetic rats were increased significantly compared with those of both acute diabetic rats and the controls. Plasma NT levels of the three groups were not different. Plasma T-SH levels of acute diabetics were increased significantly compared with those of the controls while T-SH increase in the chronic diabetics was not significant. Plasma LHP levels were increased significantly in the chronic diabetic rats compared with those of the controls. The increase in plasma PCO, AOPP, LHP levels in chronic but not in acute diabetic rats may be indicating that persistence of hyperglycemia is involved in the evolution of OPD while plasma NT levels do not seem to reflect OPD in diabetes. PMID- 12911869 TI - Prevalence of gingivitis in 6- to 11-year-old Croatian children. AB - OBJECTIVE: The purpose of this study was to assess the prevalence of gingivitis in 6- to 11-year-old children in two urban settlements of Zagreb, Croatia. PARTICIPANTS: Total number of 1025 children was included in the study. The Gingival Bleeding Index was used to determine the condition of the gingiva on mesiobuccal and buccal surfaces of six selected teeth, totaling to 12 surfaces. RESULTS: Forty-three percent of the examined children had healthy gingival tissue, 12% showed bleeding from one surface, 6% from two, 5% from three, 17% from four, and 17% from five to eight surfaces. CONCLUSIONS: Gingivitis was prevalent, but slight to moderate. It is important to instruct and control schoolchildren in excellent oral hygiene in order to prevent gingivitis. PMID- 12911870 TI - Oral manifestations of triple A syndrome. AB - Triple A (four A; Allgrove syndrome) syndrome is a an autosomal recessive disorder characterized by alacrimia, achalasia, adrenocortical insufficiency, and various neurological abnormalities. We report a case of triple A syndrome in a 14 year-old female patient together with oral manifestations. The importance of this case report is to highlight the necessity of referring the young patients with xerostomia from dental to medical specialists. PMID- 12911871 TI - Cardiovascular risk factors and probability for cardiovascular events in HIV infected patients: part I. Differences due to the acquisition of HIV-infection. AB - OBJECTIVE: Due to side effects, such as hyperlipidaemia and insulin resistance, there is an increasing concern that antiretroviral drugs lead to an epidemic of cardiovascular diseases in HIV-infected patients. The present study characterizes the cardiovascular risk profile of HIV-infected individuals, due to the acquisition of HIV-infection. METHODS: Cardiovascular risk factors of 309 HIV infected adults (HIV-acquisition: 59.2% by homosexual contact (group 1), 28.5% by heterosexual contact (group 2), 9.1% by intravenous drug abuse (group 3) and 3.2% by blood transfusion (group 4)) were analysed. Overall 10-years probability for cardiovascular events was analysed by the Framingham algorithm. RESULTS: Tobacco use was more common in group 1 subjects compared with group 2 subjects (67% vs. 52%; p<0.05). Additionally, group 1 subjects exhibited elevated total cholesterol (5.6 +/- 0.1 mmol/L vs. 4.8 +/- 0.3), LDL-cholesterol (3.6 +/- 0.1 mmol/L vs. 2.8 +/- 0.2) and triglyceride concentrations (3.2 +/- 0.3 mmol/L vs. 1.7 +/- 0.2) compared with group 3 (all p<0.05). No significant differences between the groups were detected in glucose metabolism. The overall 10-years probability for cardiovascular events was significantly higher in group 1 compared with group 2 and group 3 (12.2 +/- 0.8% vs. 6.6 +/- 0.9% and 7.9 +/- 1.6%, p<0.05). CONCLUSIONS: The cardiovascular risk profile differs between subgroups of HIV infected individuals, leading to significant higher probability of cardiovascular events in group 1 subjects. The risk of premature atherosclerosis by HIV-infected individuals and therapeutic options remains to be established. PMID- 12911872 TI - Epidemiology of chronic daily headache. AB - Over the last 10 years an increasing amount of data regarding the prevalence of chronic daily headache (CDH) has been published. The economic implications of chronic daily headache have now grown in importance in view of the increasingly limited financial resources in the health care system. In addition to recording data regarding the prevalence of this disease, epidemiological studies have also dealt with analysing and evaluating the quality of life of the afflicted patients. According to population-based data from the USA, Europe and Asia, approx. 4-5% of the population suffer from chronic daily headache. These have been equated up until now with chronic tension-type headache (CTTH). More recent epidemiological studies have resulted in an adaptation of this point of view. Currently it is assumed that approx. 2-3% of the population suffer CTTH, which preferably affects females (approximately twice as frequently); approx. 2 % suffer chronic migraine (transformed migraine = TM) and 0.2 % are afflicted with a so-called new daily persistent headache or very rarely a hemicrania continua. PMID- 12911873 TI - Dental treatment concepts for pregnant patients--results of a survey. AB - AIMS AND METHODS: 702 dentists in private practices were surveyed about dental treatments of pregnant patients with an anonymous questionnaire. RESULTS: 61.3% of the dentists indicated that they treat pregnant patients, 35.5% postponed treatment to a postnatal time if possible and 3.4% referred these patients to dental clinics. The most frequently occurring treatments measures were pain and emergency treatments in 53.3% of the cases, followed by restorative treatments (42.9%) and oral hygiene measures (39.7%). Only 9.8% of the dentists performed all necessary treatments, 14% were against using an anesthetic during pregnancy. Less than 50% of the dentists indicated they would not perform a treatment in the first trimester and 8.5% said they would not treat in the second trimester. CONCLUSIONS: The present study shows that only 54.6% of the surveyed dentists felt they were sufficiently informed and educated about the treatment of pregnant patients. PMID- 12911874 TI - Relationship between ACE gene polymorphism and ischemic chronic heart failure in Turkish population. AB - BACKGROUND: Angiotensin-converting enzyme (ACE) gene insertion/deletion (I/D) polymorphism has been associated with the development of left ventricular hypertrophy, myocardial infarction, and remodeling. However, little is known about its role in ischemic chronic heart failure (CHF). We investigated the relationship between ACE gene I/D polymorphism and ischemic CHF and its influence on exercise capacity. METHODS: ACE gene I/D polymorphism was analyzed in 209 Turkish patients with coronary artery disease (CAD) undergoing coronary angiography. ACE genotype distributions were examined in 84 consecutive patients with ischemic CHF, functional capacity class II-IV to New York Heart Association and left ventricular ejection fraction (LVEF) < 40% and 125 consecutive patients with stable angina pectoris and LVEF > or = 40%. Furthermore the results of the cardiopulmonary exercise testing (CPX) in each ACE genotype were compared in medically treated ischemic CHF patients (n = 84). RESULTS: ACE genotype distributions were similar between the patients with and without symptomatic CHF in CAD. The odds ratios were 0.95 for D homozygotes (p > 0.05) and 0.98 for the D allele (p > 0.05). In patients with ischemic CHF the differences in CPX findings were statistically not significant in ACE D/D, I/D and I/I genotypes (peak oxygen consumptions 13.7 +/- 4.6; 14.6 +/- 5.1 and 14.5 +/- 5.0 ml/kg/min, respectively (p >0.05). CONCLUSIONS: In this study population, there was no evidence that ACE gene I/D polymorphism plays a role in the development of CHF in CAD or any influence on exercise capacity in treated patients with ischemic CHF. PMID- 12911875 TI - Inguinal hernia and concomitant varicocele mimicking mesh complication. AB - In males inguinal hernia and varicocele are frequent diseases with a reported incidence of 1-2 % for hernia (Klinge 2000) and up to 20 % (Mickevicius et al. 2002) for varicocele. In 1997 approximately 220,000 inguinal herniotomies were performed in Germany (Horeyseck 1997). Leading symptoms in both diseases are complaints and dragging pain in the inguinal region radiating into the testis. Surgeons treat traditionally inguinal hernia whereas varicocele is the domain of urologists. Coincident appearance of inguinal hernia and varicocele is expected to be more frequent with regard to the pathogenesis (Friedman et al. 1993; Nakada et al. 1994; Rosch et al. 2002; Rovsing 1916; Santoro et al. 2000; Wagh and Read 1972). Therefore both surgeon and urologist should see patients with inguinal hernia or varicocele. If patients with hernia repair postoperatively suffer from similar complaints again, it should not be interpreted rashly as mesh complication (Conze et al. 2001). The need for an improved assessment has been recognized (Kehlet et al. 2002). It sounds reasonable to assume, that quite a number of patients with complaints after successful hernia repair may suffer from an undiagnosed varicocele. We present a case of inguinal hernia with mesh implantation and concomitant varicocele as an example for mesh-unrelated postherniorrhaphy pain. PMID- 12911937 TI - Molecular markers in myeloproliferative disorders: from classification to prognosis? AB - The recent description of molecular markers in patients with myeloproliferative disorders (MPDs) has raised several questions: does the presence of multiple markers coincide in individual patients or can a patient acquire some markers selectively? Do the markers distinguish molecular categories of MPDs? Do these categories coincide with the clinically defined subgroups of MPDs: PV, ET and IMF? If not, which system of categorization is more useful to the patient and his physician, the molecular one or the clinical one, and why? The present review will summarize the current knowledge of molecular markers in MPDs and discuss today's answers to the above questions. Since our knowledge of the molecular basis of MPDs is rapidly expanding, it is my hope that this review will soon be outdated. PMID- 12911938 TI - Total registration of non-Hodgkin's lymphoma and Hodgkin's disease in Scotland: effect of deprivation and caseload on outcome. AB - All cases S16 years of age with a histological diagnosis of non-Hodgkin's lymphoma (NHL) and Hodgkin's disease (HD) presented in Scotland between 1 January 1994 and 31 December 1996 were registered prospectively in the Scotland and Newcastle Lymphoma Group database by a process of total registration. The census population of Scotland in 1996-1997 was 5.1 million. One thousand seven hundred and sixty three patients were registered with NHL and 350 patients with HD. These patients have been followed up for a median of 47 months in the case of NHL and 51 months for HD cases. Actuarial 5-year survival for adult NHL was 35% and for HD, 75%. Outcome for both NHL and HD was particularly poor in the population over 60 years with median survival of 18 months for NHL and 27 months for HD. When analysis of survival was related to degree of material deprivation using the Carstairs score a significantly poorer survival was seen for NHL with increasing deprivation that could not be explained by a different pattern of age or stage at presentation. Deprivation had no impact on incidence or survival in HD. Analysis of impact of caseload of the physician initiating therapy showed no significant difference in 5-year survival. PMID- 12911940 TI - Hodgkin's disease variant of Richter's syndrome: complete remission of the both malignancies after 14 years. AB - Richter's syndrome is defined by the occurrence of high-grade malignant lymphoma in the course of chronic lymphocytic leukemia (CLL). The prognosis is poor with a life expectancy of a few months. We report on the case of a 68-year old male who developed Hodgkin's disease 6 years after the diagnosis of stage A CLL was made. The patient received 3 cycles of combination chemotherapy according to the ABVD regimen, followed by radiotherapy. This treatment resulted in complete apparent remission of the two diseases, which was persistent after a follow-up of 14 years. The Hodgkin's disease variant of Richter's syndrome is exceptional with about 20 reported cases to date. Recent research indicates that lymphoma cells derive from leukemia. The prognosis appears far better than that for the classical non-Hodgkin Richter's syndrome. PMID- 12911939 TI - Syndecan-1 in multiple myeloma: relationship to conventional prognostic factors. AB - Syndecan-1 (CD138) mediates myeloma cell adhesion, and loss of syndecan-1 from the cell surface may contribute to myeloma cell proliferation and dissemination and influence the prognosis in patients with multiple myeloma (MM). In order to test this hypothesis, we have evaluated syndecan-1 expression on the surface of malignant plasma cells and soluble forms of syndecan-1 in the serum of 25 newly diagnosed MM patients by flow cytometry and immunosorbent assay. Soluble syndecan 1 levels were significantly higher in MM as compared to controls (P<0.001). Cellular and soluble syndecan-1 was significantly inversely correlated (r=-0.89, P<0.001). The soluble syndecan-1 was significantly higher in non- responders to chemotherapy when compared to responders (P<0.01), and in non- survivors as compared to survivors (P<0.001). In contrast, cellular syndecan-1 expression was significantly lower in non- responders when compared to responders (P<0.01), and in non- survivors as compared to survivors (P<0.05). The levels of soluble syndecan-1 increased from stage I through stage II to stage III, whereas cellular syndecan-1 expression were decreased from high levels in stage III down to a low in stage I, with a statistically significant difference (P<0.01, P<0.05, respectively). There was a significant positive correlation between soluble syndecan-1 and plasma cell count (r=0.079, P<0.001), beta2 microglobulin (r=0.85, P<0.001), serum creatinine (r=0.84, P<0.001), C-reactive protein (r=0.082, P<0.001), alkaline phosphatase (r=0.58, P<0.05) and serum calcium (r=0.77, P<0.01) and a negative correlation with hemoglobin level (r=-0.78, P<0.01), platelets count (r=-0.82, P<0.01) and Albumin level (r=-0.64, P<0.01). Cox regression analysis using soluble syndecan-1 at mean-2SD of the controls could correctly classify patient outcome in 84.0%. The addition of beta2 microglobulin to soluble syndecan-1 increased the predictability of the patients' outcome to 96.7%. We conclude that soluble syndecan-1 levels are negatively correlated to the cellular form and that high levels of soluble syndecan-1 and lower expression of cellular syndecan-1 at diagnosis are negative prognostic factors. Assessment of soluble syndecan-1 and beta2 microglobulin at diagnosis is an independent prognostic system for MM. PMID- 12911941 TI - Prognostic factors in adult acute lymphoblastic leukemia. AB - The prognosis of patients with acute lymphoblastic leukemia (ALL), treated with modern chemotherapeutic regimens, is dependent on a number of variables. The major prognostic factors for survival in adult ALL are age, cytogenetic abnormalities, immunologic subtype, white blood cell (WBC) count, and time to achieve complete remission (CR). Determination of these factors is crucial for adapting post remission therapy in adult ALL. Indeed, risk-adapted strategies based on those biologic and clinical features are currently being applied to improve survival. In this review, we report the different prognostic factors described in adult ALL and discuss the controversies in current adult ALL management in relation with these different features. The data reported are derived from the medical literature and from the experience of the authors. Prognostic factors appear to be time-dependent. This emphasizes their determination according to the phase of treatment. The use of time-segmented multivariate analysis able to distinguish prognostic factors associated with the induction phase and those associated with the post-induction phase of treatment seems suitable to define accurately prognostic models. PMID- 12911943 TI - Single center review of clinicopathological characterization in 77 patients with positive lupus anticoagulant antibodies. AB - BACKGROUND: The antiphospholipid syndrome (APS) is a thrombophillic disorder characterized by the presence of antiphospholipid antibodies (APA). It often occurs in patients with systemic lupus erythematosus (SLE) and may be associated with recurrent abortions and thrombocytopenia and, occasionally, catastrophic thrombotic events. OBJECTIVES: To examine, retrospectively, the clinico pathological features of patients with APS detected by the presence of the lupus anticoagulant (LAC). METHODS: Patients were selected for study on the basis of a positive LAC test on review of the laboratory computer records of the King Faisal Specialist Hospital and Research Center. Following this, a clinical chart review was conducted in order to determine the clinical presentations, treatment and the course of patients identified. The information obtained was entered into an electronic database and subsequently analyzed. RESULTS: Seventy-seven patients were identified and reviewed. Fifty-six (73%) were female and 16 (21%) were children less than 15-years-old. Thirty-two patients (42%) had no clinical events (incidental APS). The syndrome was classified as primary in 40 (52%) patients and secondary in 37 (48%). Out of the 45 (58%) patients who presented with symptoms related to APA 22 (49%) had thrombosis, 24 (53%) had pregnancy failure, and 4 (9%) presented with catastrophic APS. The activated partial thromboplastin time (aPTT) was elevated and not corrected by mixing with normal plasma in 47 (61%). On the other hand, the prothrombin time (PT) was normal in 66 (90%). There is a significant difference between aPTT and PT as a screening test with P value of < 0.0001. Tests for anticardiolipin antibodies (ACA) were positive in 39 patients (70%). Only 13 (17%) patients had thrombocytopenia. All patients who presented with thrombosis were treated with warfarin but only 5 (23%) had received aspirin. Out of the 22 patients presenting with thrombosis, 12 (55%) had one or more recurrent thrombotic events while only 6 (25%) out of the 24 patients who presented with pregnancy failure had events other than pregnancy failure. Fifty two patients were followed up regularly and were alive. CONCLUSIONS: We find that thrombosis, venous or arterial, and obstetric complications are the most frequent clinical findings in our patients with circulating LAC. Incidental APS is not an uncommon finding in patients screened for APS. There is a clear association between the presence of LAC and an abnormal aPTT, which is much less obvious with the PT. PMID- 12911942 TI - Evaluating the effectiveness of epoetin alfa in community oncology practices. AB - PURPOSE: This retrospective case-series review studied the effectiveness of epoetin alfa in community oncology practices, which until now has not been well documented. METHODS: We reviewed the medical records of 118 cancer patients treated between 1999 and 2001 with cyclic chemotherapy plus epoetin alfa in 27 US community-based oncology practices. Two analysis sets were examined: one including all patients (n=118) and one including only those patients with no concurrent events impacting hemoglobin data interpretation (n=73). Efficacy of epoetin alfa was evaluated by hemoglobin response (aS2 g/dl increase in hemoglobin from baseline) at weeks 12 and 16, the time to hemoglobin response, and change in hemoglobin concentrations from baseline at specific time points. RESULTS: After 12 weeks of treatment, 43% (95% confidence interval [CI], 33-54%) of patients had a hemoglobin response, and the proportion of responders further rose to 61% (95% CI, 49-72%) after 16 weeks. The median time to response was 92 days (lower 95% confidence limit, 74 days; upper bound not estimable). Hemoglobin increased from baseline at all time points evaluated during epoetin alfa treatment, with a mean increase of 1.1 g/dl (95% CI, 0.77-1.4 g/dl) by the last observation. CONCLUSION: These results indicate that epoetin alfa is effective in community practice, but most patients take longer than 3 months to respond. Because slow responses may negatively impact on the quality of life in these patients, alternative treatment approaches providing faster and perhaps better responses may provide greater clinical benefit. PMID- 12911944 TI - Platelet count increase following phlebotomy in iron overloaded patients with liver cirrhosis. AB - Thrombocytopenia is a frequent hematological complication in patients with liver cirrhosis, but its pathogenesis is not clearly understood. We evaluated the effect of iron depletion by phlebotomy on platelet count in 62 consecutive iron overloaded patients with liver cirrhosis and thrombocytopenia. After a median follow-up of 30.2 months we observed a significant increase of platelet count in all patients (from mean baseline levels of 110.1 up to 168.22109/l at the end of follow-up, P<0.001) with platelet count normalization in 42 of them (67.7%). In addition, we observed a significant improvement of serum ALT levels (from pretreatment mean values of 126.7 up to 59.7 U/l at the end of follow-up, P<0.001) along with the reduction of serum ferritin levels and transferrin saturation during phlebotomy. Different pathogenetic mechanisms involving both humoral (erythropoietin and thrombopoietin, TPO) and physical (portal hypertension and hypersplenism) factors are here discussed to explain the platelet count increase following phlebotomy. Our results show that phlebotomy is effective not only in lowering iron overload, but also in improving liver function and thrombocytopenia in patients with liver cirrhosis. PMID- 12911945 TI - Recombinant factor VIIa used to control massive haemorrhage during renal transplantation surgery; vascular graft remained patent. AB - There is a growing body of evidence to support the use of rFVIIa in cases of excessive bleeding outside its approved use in haemophilia with inhibitors. We now report its use for a paediatric renal transplant case in which its use was both life saving and did not threaten the vascular grafts. To our knowledge this represents the first reported case of rFVIIa used in the setting of renal transplantation. It is reassuring that despite use of a high dose of rFVIIa (>135 mcg/kg; recommended standard dose is 90 mcg/kg) there was no evidence of anastomotic thrombosis. PMID- 12911946 TI - Assessment of neutrophil apoptosis ex vivo in hepatosplenic patients with neutropenia pre and post splenectomy. AB - The pathophysiology of neutropenia seen in patients with schistosomiasis or hepatitis C infection that complicates the course of liver disease is poorly understood. We evaluated the neutrophil apoptosis before and after splenectomy to clarify the role of apoptosis and splenomegaly in the occurrence of neutropenia. Neutrophils were isolated from 23 hepato-splenic patients with neutropenia, 8 hepatosplenic patients with normal neutrophil counts, 7 patients who were post splenectomy, and a further ten normal control subjects. These were cultured for 24 h and the time course of neutrophil apoptosis was assessed by determination of Annexin V and propidium iodide binding by flow cytometry. Fas and Bcl2 expression were determined on fresh neutrophils using flow cytometry. Levels of tumor necrosis factor alpha, interleukin 3, and gamma interferon were evaluated using an immunosorbent assay. Neutrophil apoptosis was minimal in the fresh neutrophils, however, cultured neutrophils exhibited significantly greater apoptosis in neutropenic patients when compared to non-neutropenic patients (P=0.01 at 4 h and P<0.05 at 24 h) and control group (P<0.01 at 4 h and 24 h). After splenectomy, the percentage of neutrophil apoptosis declined to the normal control levels (P>0.05). Fas and Bcl2 expression on neutrophil were significantly higher in the neutropenic group as compared to normal controls (P<0.05, P=0.01 respectively). Serum TNF alpha, IL-3, and IFN gamma levels were not significantly different in all studied groups. IN CONCLUSION: Neutrophils from neutropenic hepatosplenic patients exhibit markedly accelerated apoptosis, which is normalized after splenectomy. Thus increased neutrophil apoptosis may in part be responsible for the occurrence of neutropenia. PMID- 12911947 TI - Hematology morphology forum case 3. Chediak-Higashi syndrome. PMID- 12912684 TI - Risk factors for various causes of failure in initial corneal grafts. AB - OBJECTIVE: To determine the risk factors for specific causes of initial corneal graft failure. METHODS: This study analyzed corneal graft survival rates in a longitudinal noncomparative case series of 3992 consecutive penetrating keratoplasties performed at a single large referral center. Regrafts (n = 352) were excluded from the analysis of risk factors for initial graft failure. Data were collected retrospectively from August 1, 1982, through December 31, 1986, and prospectively from January 1, 1987, through August 31, 1996. Patients were examined preoperatively, at 1, 3, 6, 9, 12, 18, and 24 months posttransplantation, and annually thereafter. Potential risk factors were evaluated individually by Kaplan-Meier survival analysis. Cox proportional hazards regression modeling was then used to investigate the impact of each independent variable, adjusted for the confounding influence of the other independent variables. RESULTS: The use of topical glaucoma medications was a significant risk factor for corneal graft failure by 3 major causes: rejection, endothelial decompensation without a documented immunologic reaction, and ocular surface disease. Deep stromal vascularization was an independent risk factor for rejection failure. Diabetes mellitus, peripheral anterior synechiae, recipient race, and small trephination size were significant risk factors for endothelial failure. CONCLUSION: Independent risk factors differentially impact specific causes of corneal graft failure. PMID- 12912685 TI - Optic disc traction syndrome associated with central retinal vein occlusion. AB - OBJECTIVE: To describe the association between ischemic central retinal vein occlusion (CRVO) and the development of optic disc (vitreopapillary) traction, as verified by optical coherence tomography (OCT). METHODS: In a prospective, noncomparative, observational patient series, 3 women aged 51 to 64 years developed an ischemic type of CRVO. One patient underwent cannulation of the central retinal vein with injection of a tissue plasminogen activator. In each eye, the contour of the optic nerve head could not be accurately detected because of overlying fibrous tissue. Each patient underwent OCT examination 6 to 10 months (average, 8.3 months) after the occlusive event. RESULTS: Optic disc traction was found by OCT in the 3 patients. In each, the disc was elevated, associated with either incomplete posterior vitreous detachment (2 eyes) or vitreopapillary fibrous membrane (1 eye). Secondary peripapillary retinal traction and macular or retinal detachment developed ("optic disc traction syndrome"). These findings were less marked, or not evident, on both clinical examination and ocular ultrasonography. CONCLUSIONS: Optic disc traction and secondary localized retinal detachment can develop after ischemic CRVO and may contribute to the poor visual acuity. This syndrome should merit special attention before surgery for CRVO is planned. The use of OCT was helpful to diagnose this syndrome. PMID- 12912686 TI - Maculas affected by age-related macular degeneration contain increased chelatable iron in the retinal pigment epithelium and Bruch's membrane. AB - OBJECTIVE: To investigate whether iron is involved in the pathogenesis of age related macular degeneration (AMD). METHODS: Postmortem AMD-affected (nonexudative or exudative) and healthy maculas were studied using the 3,3' diaminobenzidine-enhanced Perls Prussian blue stain. The Perls Prussian blue stain was quantified by computer-assisted analysis of digital images. To determine whether the iron was chelatable, sections treated with the iron chelator deferoxamine were compared with adjacent, nonchelated sections. RESULTS: Compared with healthy maculas, AMD-affected maculas had statistically significant increases in the total iron level. Some of this iron was chelatable. The iron was present in retinal pigment epithelium and Bruch's membrane in maculas from patients who had drusen only, geographic atrophy, and exudative AMD in pathologic areas and, occasionally, in relatively healthy areas. CONCLUSIONS: Oxidative stress has been implicated in the pathogenesis of AMD by the Age-Related Eye Disease Study. Increased concentrations of iron, which generate highly reactive hydroxyl radicals via the Fenton reaction, may induce oxidative stress in the macula and lead to AMD. As the increased iron concentrations in AMD-affected eyes consist in part of a chelatable iron pool, treatment of patients who have AMD with iron chelators might be considered a potential therapy. While there are, as yet, no clinical data indicating that the treatment of patients who have AMD with iron chelators is beneficial, data presented herein indicate that further investigation of iron concentrations in postmortem tissues and the mechanisms of iron transport in the retina is warranted. PMID- 12912687 TI - Clinical description and exclusion of candidate genes in a novel autosomal recessively inherited vitreoretinal dystrophy. AB - OBJECTIVES: To describe the clinical phenotype of a novel autosomal recessively inherited vitreoretinal dystrophy in one generation of a family originating from eastern Switzerland. METHODS: A clinical study including electroretinographic investigations followed by laboratory-based genetic and molecular analysis. Four affected and 3 unaffected members of the family were examined. Ten candidate regions were tested by linkage analysis with highly polymorphic molecular markers or with intragenic restriction fragment length polymorphisms. RESULTS: Of 8 siblings,4 were affected, showing high myopia with pronounced vitreous liquefaction, retinitis pigmentosa-like retinal degeneration, diffuse retinal pigment epithelium atrophy, macular staphylomata, and premature cataract formation. Strikingly abnormal results on electroretinograms, affecting both the rod and the cone systems, revealed an extensive defect of retinal function, unlike those usually found in pathologic myopia. No extraocular manifestations were observed. Three types of nonsyndromic high myopia, Stickler syndrome I, II, and III, Wagner syndrome, Knobloch syndrome, Goldmann-Favre dystrophy, and multiple vitreoretinopathies were excluded by linkage analysis. CONCLUSIONS: The reported phenotype as well as the results of molecular linkage analysis in the siblings described here suggest an autosomal recessively inherited vitreoretinal dystrophy, which, to our knowledge, has not been described until now. PMID- 12912688 TI - Prognostic value of S-100-beta serum concentration in patients with uveal melanoma. AB - BACKGROUND: In cutaneous melanoma, the S-100-beta serum level is recognized as a marker of metastatic disease. OBJECTIVES: To determine whether S-100-beta is present in the serum of patients with uveal melanoma and to test whether the serum concentration of S-100-beta is related to known clinical and histopathological prognostic factors in these patients. METHODS: The S-100-beta concentration was measured in serum samples collected from 64 patients with uveal melanoma before enucleation and from 58 healthy control subjects. A 2-site immunoluminometric assay was used to quantify the S-100-beta concentration in serum. S-100-beta concentrations in the serum from patients were compared with clinicopathological tumor variables, sex, occurrence of metastasis, and survival. RESULTS: Thirty-seven (57.8%) of 64 patients with uveal melanoma showed detectable levels of serum S-100-beta. There was, however, no significant difference between serum levels of patients and control subjects (P =.71). Statistical analysis showed no significant correlation between S-100-beta concentration and any of the clinicopathological tumor variables, occurrence of metastases, or survival. Only sex was correlated with S-100-beta serum levels, which was not observed in the control group. CONCLUSIONS: In our study on patients with uveal melanoma, the S-100-beta serum concentration was not correlated with any investigated prognostic factor and was not of prognostic value itself. Female patients appeared to have higher S-100-beta concentrations than male patients. PMID- 12912689 TI - Ocular motility changes after subtenon carboplatin chemotherapy for retinoblastoma. AB - BACKGROUND: Focal subtenon carboplatin injections have recently been used as a presumably toxicity-free adjunct to systemic chemotherapy for intraocular retinoblastoma. OBJECTIVE: To report our clinical experience with abnormal ocular motility in patients treated with subtenon carboplatin chemotherapy. METHODS: We noted abnormal ocular motility in 10 consecutive patients with retinoblastoma who had received subtenon carboplatin. During ocular manipulation under general anesthesia, we assessed their eyes by forced duction testing, comparing ocular motility after tumor control with ocular motility at diagnosis. Eyes subsequently enucleated because of treatment failure (n = 4) were examined histologically. RESULTS: Limitation of ocular motility was detected in all 12 eyes of 10 patients treated for intraocular retinoblastoma with 1 to 6 injections of subtenon carboplatin as part of multimodality therapy. Histopathological examination revealed many lipophages in the periorbital fat surrounding the optic nerve in 1 eye, indicative of phagocytosis of previously existing fat cells and suggesting prior fat necrosis. The enucleations were technically difficult and hazardous for globe rupture because of extensive orbital soft tissue adhesions. CONCLUSIONS: Subtenon carboplatin chemotherapy is associated with significant fibrosis of orbital soft tissues, leading to mechanical restriction of eye movements and making subsequent enucleation difficult. Subtenon carboplatin is not free of toxicity, and its use is best restricted to specific indications. PMID- 12912690 TI - Histopathologic findings in eyes with retinoblastoma treated only with chemoreduction. AB - OBJECTIVE: To evaluate the histopathologic findings in the eyes with retinoblastoma that had been treated only with chemoreduction. DESIGN: Clinicopathologic series. Study Material Ten eyes of 8 patients with retinoblastoma that were enucleated after therapy consisting only of systemic chemotherapy (chemoreduction). METHODS: All cases received a chemoreduction regimen including a combination of intravenous carboplatin, etoposide phosphate, and vincristine sulfate. Adjuvant treatment to the tumor was not provided in any case. The enucleated globes were studied by routine light microscopy. Main Outcome Measure Histopathologic features of retinoblastoma following chemoreduction. RESULTS: At presentation, there were 8 eyes in Reese-Ellsworth group V, 1 eye in group IV, and 1 eye in group III. After chemoreduction (mean, 4 cycles; range, 1-6 cycles), the main tumor regressed a mean 34% in thickness and 24% in basal diameter. The indication for enucleation was retinoblastoma recurrence as subretinal and/or vitreous seeds in 7 eyes and extensive vitreous hemorrhage in 3 with uncertainty about viable-appearing tumor. In no case was enucleation performed for recurrence of the main tumor. In all eyes, there was histopathologic evidence of tumor regression. In 8 of 10 eyes, histopathologic examination disclosed tumor regression without viable-appearing retinoblastoma in the main tumor. Of these 8 eyes, 2 showed a completely calcified glial scar and 6 showed an apical calcified glial scar and a basal residual well-differentiated component with retinomalike and/or retinocytomalike features. In the remaining 2 eyes, an area of posttherapeutic regression was present but contained foci of mitotically active, viable-appearing malignant retinoblastoma cells. The 6 eyes found to contain well-differentiated component with retinomalike and/or retinocytomalike features showed a mean decrease of 17% in largest basal dimension and 32% in thickness after a mean of 3 cycles of chemoreduction. In contrast, the 4 eyes that did not contain well-differentiated component with retinomalike and/or retinocytomalike features showed a mean decrease of 35% in largest basal dimension and 55% in thickness after a mean of 5 cycles of chemoreduction. Of those 7 eyes enucleated for recurrent subretinal and/or vitreous seeds, viable tumor seeds were confirmed histopathologically in all cases. There was no histopathologic evidence of chemotherapeutic toxicity to the eye. CONCLUSIONS: Histopathologic examination of 10 enucleated eyes following chemoreduction alone revealed that the main retinoblastoma regressed in all eyes. Additionally 6 eyes showed basal residual well-differentiated component with retinomalike and/or retinocytomalike features, and these eyes also displayed less shrinkage with chemoreduction. Despite the lack of viable-appearing retinoblastoma within the main tumor, enucleation was performed for viable subretinal and/or vitreous seeds in 7 cases and confirmed histopathologically. PMID- 12912691 TI - Alterations in corneal stromal dendritic cell phenotype and distribution in inflammation. AB - BACKGROUND: The normal corneal stroma is endowed with large numbers of resident dendritic cells (DCs). The purpose of this study was to examine the phenotype and distribution of these cells in inflammation. METHODS: Normal and inflamed murine corneas were excised at different time points and immunofluorescence staining with multiple antibodies was performed by confocal microscopy on whole-mounted corneal stromas to characterize and evaluate the distribution of DCs in inflammation. RESULTS: CD11c+CD11b+ myeloid DCs were present throughout the anterior stroma. In the periphery of the normal cornea, nearly one half were major histocompatibility complex (MHC) class II+CD80+CD86+, while they were uniformly MHC class II-CD80-CD86- in the center. In inflammation, in addition to a significant increase in the number of DCs, a majority up-regulated their expression of MHC class II, CD80, and CD86, indicating their state of maturation. The up-regulation of MHC class II and costimulatory molecules on DCs was seen as early as 24 hours after induction of inflammation or transplantation. In addition to the CD11c+DCs in the anterior stroma, a CD11c-CD11b+ population of monocytes/macrophages was present almost exclusively in the posterior stroma of the cornea. These cells were found throughout all layers of the stroma, and in increased numbers, after induction of inflammation. CONCLUSION: The present study demonstrates, for the first time, the phenotypic changes and distribution of resident stromal DCs in corneal inflammation. Clinical Relevance These novel data suggest that the cornea is capable of participating in immune and inflammatory responses by virtue of its own heterogeneous population of DCs. PMID- 12912692 TI - Refractive error and ethnicity in children. AB - OBJECTIVE: To report the baseline prevalence of refractive error in the study population. DESIGN: A multicenter, longitudinal, observational study of refractive error and ocular development in children from 4 ethnic groups. PATIENTS AND METHODS: The study population included 2523 children (534 African American, 491 Asian, 463 Hispanic, and 1035 white) in grades 1 to 8 (age, 5-17 years). Myopia was defined as -0.75 diopters (D) or more and hyperopia as +1.25 D or more in each principal meridian, and astigmatism was defined as at least a 1.00-D difference between the 2 principal meridians (cycloplegic autorefraction). RESULTS: Overall, 9.2% of the children were myopic, 12.8% were hyperopic, and 28.4% were astigmatic. There were significant differences in the refractive error prevalences as a function of ethnicity (chi2, P<.001), even after controlling for age and sex (polychotomous logistic regression, P<.001). For myopia, Asians had the highest prevalence (18.5%), followed by Hispanics (13.2%). Whites had the lowest prevalence of myopia (4.4%), which was not significantly different from African Americans (6.6%). For hyperopia, whites had the highest prevalence (19.3%), followed by Hispanics (12.7%). Asians had the lowest prevalence of hyperopia (6.3%) and were not significantly different from African Americans (6.4%). For astigmatism, Asians and Hispanics had the highest prevalences (33.6% and 36.9%, respectively) and did not differ from each other (P =.17). African Americans had the lowest prevalence of astigmatism (20.0%), followed by whites (26.4%). CONCLUSION: There were significant differences in the prevalence of refractive errors among ethnic groups, even after controlling for age and sex (P<.001). PMID- 12912693 TI - Relation of statin use to the 5-year incidence and progression of age-related maculopathy. AB - OBJECTIVE: To examine the association of hydroxymethyl glutaryl coenzyme A reductase inhibitors (statins) with the 5-year incidence of age-related maculopathy (ARM). DESIGN: Population-based cohort study. Participants included persons 48 to 91 years old examined March 1, 1993, through June 14, 1995, living in Beaver Dam, Wis (N = 3684), of whom 2780 participated in a follow-up 5 years later. METHODS: Standardized procedures were used for physical examinations, blood sample collection, and questionnaire administration. Age-related maculopathy was determined by grading images of the posterior pole using a standard protocol. Standard univariate and multivariate analyses were performed. MAIN OUTCOME MEASURES: Incidence and progression of ARM was measured over the 5 year interval. RESULTS: While controlling for age and sex, statin use was not found to be associated with the 5-year incidence of early ARM (odds ratio [OR], 1.12; 95% confidence interval [CI], 0.47-2.67), progression of ARM (OR, 1.22; 95% CI, 0.54-2.76), or incidence of late ARM (OR, 0.41; 95% CI, 0.12-1.45). CONCLUSIONS: These findings do not suggest an association between statin use and incident ARM over a 5-year period. Further investigation of these relationships in larger studies over a longer period is needed. PMID- 12912694 TI - Trends in size and treatment of recently diagnosed choroidal melanoma, 1987-1997: findings from patients examined at collaborative ocular melanoma study (COMS) centers: COMS report no. 20. AB - OBJECTIVE: To describe time trends, from 1987 through 1997, (1) in size of choroidal melanoma among patients with recent diagnosis confirmed at a clinical center that participated in the Collaborative Ocular Melanoma Study (COMS) and (2) in choice of treatment by patients who did not enroll in one of the COMS randomized trials. METHODS: Investigators at all COMS clinical centers (41 in the United States and 2 in Canada) agreed to report, in a masked fashion that did not include personal identifiers, all patients diagnosed as having choroidal melanoma during the accrual period for COMS randomized trials of radiotherapy. Information reported for patients who did not enroll in a COMS randomized trial included tumor dimensions, date of diagnosis, and initial treatment selected. Patients reported by centers that continued to report cases until 1997 and diagnosed as having choroidal melanoma no more than 1 year before evaluation at a participating COMS center contributed the data analyzed. OUTCOME MEASURES: Time trends in tumor size among patients reported and in elective treatment of patients not enrolled in COMS randomized trials. RESULTS: Of 8712 patients with choroidal melanoma examined, 6703 met criteria for analysis of time trend in tumor size and 4077 were analyzed for treatment trends over time. The number of cases with longest tumor basal diameter greater than 15.0 mm declined over time from 393 (30%) of 1330 cases reported in 1987 through 1989 to 345 (25%) of 1397 cases reported in 1996 or 1997. The proportion of patients eligible for COMS randomized trials who did not enroll and who elected enucleation remained stable over time for tumors of all sizes; the proportion of these patients who elected eye-conserving radiotherapy increased over time. Juxtapapillary tumors accounted for nearly half of the enucleations among ineligible patients who had tumors no larger than 15.0 mm in longest basal diameter. CONCLUSIONS: Among patients examined at COMS centers during 1987 through 1997, the trends observed for patients with recently diagnosed choroidal melanoma included increasing presentation with tumors of smaller size, decreasing use of enucleation for choroidal melanoma 15.0 mm or less in longest basal diameter, and increasing use of such strategies for larger tumors. PMID- 12912695 TI - Comparison of clinical, echographic, and histopathological measurements from eyes with medium-sized choroidal melanoma in the collaborative ocular melanoma study: COMS report no. 21. AB - OBJECTIVE: To compare pre-enucleation clinical and echographic measurements with postenucleation histopathological measurements of choroidal melanoma of a size and in a location suitable for iodine 125 brachytherapy. DESIGN: Cross-sectional study of patients with melanoma classified as medium-sized who were assigned to the enucleation arm of the randomized trial of 125I brachytherapy vs enucleation conducted by the Collaborative Ocular Melanoma Study (COMS) Group. METHODS: Measurements of melanoma dimensions by clinical, echographic, and histopathological examinations were compared for 644 eyes. Outcomes Tumor longest basal diameter and apical height. RESULTS: Tumor dimensions were available for 644 (98%) of 660 patients who had unilateral medium-sized choroidal melanoma and were randomly assigned to enucleation. The clinical and histopathological measurements of the longest basal diameter agreed within +/-2 mm for 371 eyes (58%). The clinical measurement was less than the histopathological measurement by more than 2 mm in 32 eyes (5%), which occurred more frequently when the tumor was within 2 mm of the optic disc. The echographic and histopathological measurements of apical height agreed within +/-2 mm in 579 eyes (90%); the echographic measurement was less than the histopathological measurement in only 2 cases. Features associated with echographic estimates larger than histopathological measurements were greater tumor height and anterior tumor apex location. CONCLUSIONS: Our results suggest that tumor measurements made according to COMS protocol were highly reliable in planning radioactive plaque therapy and monitoring changes in tumor size after such treatment. PMID- 12912696 TI - Glaucoma phenotype in pedigrees with the myocilin Thr377Met mutation. AB - OBJECTIVE: To investigate the phenotype and age-related penetrance of primary open-angle glaucoma (POAG) in Australian families with the myocilin mutation Thr377Met. METHOD AND DESIGN: Cross-sectional genetic study. Four unrelated pedigrees carrying the Thr377Met mutation were ascertained from more than 2000 consecutive cases of POAG in the Glaucoma Inheritance Study in Tasmania and from families with glaucoma referred to the study from throughout Australia. Index cases and available family members were examined for signs of glaucoma, and the presence of the GLC1A Thr377Met mutation was ascertained by single-strand conformation polymorphism analysis and subsequent direct sequencing. RESULTS: From the 4 pedigrees carrying the Thr377Met mutation, 23 individuals with either ocular hypertension (OHT) or POAG were found, with a mean +/- SD age at diagnosis of 41.2 +/- 11.5 years, and a mean peak intraocular pressure of 31.7 +/- 9.9 mm Hg. A further 9 mutation carriers older than 18 years were studied who as yet showed no signs of OHT or POAG (6 of these 9 were younger than 30 years). A single individual with POAG was identified who did not carry the Thr377Met mutation. For Thr377Met carriers, age-related penetrance for OHT or POAG was 88% at age 30 years. A positive family history of POAG was present for 3 of the 4 index cases. Thirteen (57%) of the 23 Thr377Met carriers with OHT or POAG had undergone glaucoma drainage surgery. Although the glaucoma in these families appears to be pressure dependent, 2 individuals showed optic disc cupping before detected elevation in intraocular pressure. One family was of British origin, with a different background haplotype from the other 3 families from Greece or Macedonia, who shared a common haplotype. CONCLUSIONS: The GLC1A Thr377Met mutation is associated with POAG that, in the pedigrees studied, had a younger age at onset and higher peak intraocular pressure than in pedigrees with the more common Gln368STOP mutation. In addition, patients with glaucoma with the Thr377Met mutation were more likely to have undergone glaucoma drainage surgery. PMID- 12912697 TI - Lack of association of mutations in optineurin with disease in patients with adult-onset primary open-angle glaucoma. AB - OBJECTIVE: To determine whether mutations in the optineurin gene contribute to susceptibility to adult-onset primary open-angle glaucoma. METHODS: The optineurin gene was screened in 86 probands with adult-onset primary open-angle glaucoma and in 80 age-matched control subjects. Exons 4 and 5, containing the recurrent mutations identified in patients with normal-tension glaucoma, were sequenced in all individuals studied, while the remaining exons were screened for DNA sequence variants with denaturing high-performance liquid chromatography. RESULTS: The recurrent mutation, Met98Lys, previously found to be associated with an increased risk of disease was found in 8 (9%) of 86 probands. We also found the Met98Lys mutation in 10% of individuals from a control population of similar age, sex, and ethnicity. Consistent segregation of the mutation with the disease was not demonstrated in any of the 8 families. No other DNA changes altering the amino acid structure of the protein were found. CONCLUSION: The mutations in the optineurin gene associated with normal-tension glaucoma are not associated with adult-onset primary open-angle glaucoma in this patient population. Clinical Relevance Genetic abnormalities that render the optic nerve susceptible to degeneration are excellent candidates for genetic factors that could contribute to adult-onset primary open-angle glaucoma. Mutations in optineurin have been associated with normal-tension glaucoma, but are not associated with disease in patients with adult-onset primary open-angle glaucoma. This result may indicate that normal-tension glaucoma is not necessarily part of the phenotypic spectrum of adult open-angle glaucoma. PMID- 12912699 TI - Microsurgical approach to the conjunctival flap. AB - OBJECTIVES: To describe a microsurgical approach to a selective pedunculated conjunctival flap for the treatment of chronic corneal ulceration with or without corneal perforation, and to present the results of 50 consecutive cases. DESIGN: Retrospective, noncomparative case series and case reports. Patients All patients who had microsurgical conjunctival flap procedures for the treatment of chronic corneal ulcers between 1982 and 1996 at the Connecticut Eyecare Center, New Haven. INTERVENTIONS: Partial pedunculated conjunctival flap surgery. METHODS: Review of the initial ocular diagnoses and characteristics as well as retrospective study of the postoperative course. MAIN OUTCOME MEASURES: Resolution of the corneal ulcer and postoperative stability of the conjunctival flap. RESULTS: Sixty-two percent of the corneal ulcers treated were nonperforated, and 38% were perforated. The diagnoses included herpes simplex virus (14 patients), bacterial ulcer (11 patients), rheumatoid arthritis (8 patients), aphakic bullous keratopathy (6 patients), graft rejection (4 patients), herpes zoster virus (5 patients), and other (2 patients). Postoperatively, 94% of the conjunctival flaps were stable, and 3% had failed. The procedures were definitive in 64% of cases and temporary in 36%; 61% of patients received a corneal transplant 6 to 24 months postoperatively, and in 38% the flaps were removed 6 to 12 months after the original lesion had healed. CONCLUSION: A selective pedunculated conjunctival flap is an effective and practical surgical approach to the treatment of perforated and nonperforated corneal ulcers that have not responded to other types of medical therapy. PMID- 12912700 TI - Epidemiology, ethnicity, race, and risk. PMID- 12912698 TI - Genomewide homozygosity mapping and molecular analysis of a candidate gene located on 22q13 (fibulin-1) in a previously undescribed vitreoretinal dystrophy. AB - OBJECTIVES: To localize the gene that causes an autosomal recessively inherited vitreoretinal dystrophy that has not been described, to our knowledge, and to analyze a candidate gene mapped to 22q13 (fibulin-1 [FBLN1]). METHODS: Homozygosity mapping with 500 microsatellite markers spread over the whole genome (mean distance, 7.2 centimorgans [cM]) and mutation analysis of the complete coding region of FBLN1. RESULTS: Homozygosity for all analyzed markers was found in the 4 affected siblings in a region on chromosome 22 encompassing 12 cM from D22S444 (centromeric) to D22S1170 (telomeric). Lod scores were between 0.017 and 2.36 (theta = 0). A mutation analysis of the complete coding region of FBLN1, which encodes interacting extracellular matrix proteins, revealed 4 previously undescribed single nucleotide polymorphisms. CONCLUSIONS: A genomewide homozygosity mapping analysis supported the hypothesis that the gene responsible for a unique vitreoretinal dystrophy is located on chromosome 22q13. No obviously pathogenic mutation was found in the candidate gene, FBLN1. PMID- 12912701 TI - Gouty tophus at the lateral canthus. PMID- 12912702 TI - Pyogenic granuloma of the cornea in an infant with unilateral microphthalmia. PMID- 12912703 TI - Congenital toxoplasmosis with unusual retinal findings. PMID- 12912704 TI - The utility of 0.5% apraclonidine in the diagnosis of horner syndrome. PMID- 12912705 TI - Myelination of the macula associated with disabling photophobia. PMID- 12912706 TI - Bilateral neuroretinopathy with multiple retinal arterial aneurysms. PMID- 12912707 TI - Ophthalmoscope or Augenspiegel? PMID- 12912708 TI - Aortic stenosis: a new face for an old disease. PMID- 12912709 TI - Key clinical, ethical, and policy issues in the evaluation of the safety and effectiveness of solid organ transplantation in HIV-infected patients. PMID- 12912710 TI - Benefits of combination therapy of insulin and oral hypoglycemic agents. PMID- 12912711 TI - Insulin and oral hypoglycemic agents should not be used in combination in the treatment of type 2 diabetes. PMID- 12912713 TI - Cardiovascular risk factors and increased carotid intima-media thickness in healthy young adults: the Atherosclerosis Risk in Young Adults (ARYA) Study. AB - BACKGROUND: The relation between cardiovascular risk factors and extent of atherosclerosis in middle-aged and elderly populations is well established. Autopsy studies have suggested that similar associations may be present at a young age. We evaluated the relationship between conventional risk factors and increased carotid intima-media thickness (CIMT) in 750 healthy young adults, aged 27 to 30 years. METHODS: All participants completed a questionnaire on cardiovascular risk factors, had a fasting blood sample drawn, and underwent an ultrasonographic examination of both common carotid arteries to assess common CIMT (CCIMT). RESULTS: Age (11.2 micro m/SD), body mass index (10.3 micro m/SD), pulse pressure (5.0 micro m/SD), sex (4.8 micro m/SD), and low-density lipoprotein cholesterol level (4.3 micro m/SD) were independent determinants of increased CCIMT in young adults (R2 = 0.36). Total pack-years of smoking, adjusted for age and sex, showed a linear trend with increased CCIMT (P =.02), which attenuated after further adjustment for body mass index. Common CIMT increased gradually and significantly with the number of cardiovascular risk factors present. The estimated absolute risk, based on the Framingham risk function, for development of coronary heart disease within 20 years was 2.5 times higher in individuals with mean CCIMT in the highest quartile compared with those in the lowest quartile of the distribution. CONCLUSIONS: An unfavorable cardiovascular risk profile is associated with a marked increase in CCIMT in young adulthood. Efforts to change modifiable risk factors early in life may retard atherosclerosis development and hence delay the onset of clinical cardiovascular disease later in life. PMID- 12912714 TI - Effect of amoxicillin-clavulanate in clinically diagnosed acute rhinosinusitis: a placebo-controlled, double-blind, randomized trial in general practice. AB - BACKGROUND: Acute rhinosinusitis is one of the most common reasons for prescribing antibiotics in primary care. However, it is not clear whether antibiotics improve the outcome for patients with clinically diagnosed acute rhinosinusitis. We evaluated the effect of a combination product of amoxicillin potassium clavulanate on adults with acute rhinosinusitis that was clinically diagnosed in a general practice setting. METHODS: We conducted a randomized, placebo-controlled, double-blind trial with 252 adults recruited at 24 general practices and 2 outpatient clinics. Each patient had a history of purulent nasal discharge and maxillary or frontal pain for at least 48 hours. Patients were given amoxicillin, 875 mg, and clavulanic acid, 125 mg, or placebo twice daily for 6 days. Main outcome measures were time to cure (primary outcome), number of days during which rhinosinusitis restricted activities at home or work, and frequency of adverse effects (secondary outcomes). RESULTS: The adjusted hazard ratio for the effect of amoxicillin-clavulanate was 0.99 (95% confidence interval [CI], 0.68-1.45) on time to cure and 1.28 (95% CI, 0.80-2.05) in the prespecified subgroup of patients with a positive rhinoscopy result. At 7 days the mean difference between amoxicillin-clavulanate and placebo was -0.29 (95% CI, -0.93 to 0.34) in the number of days with restrictions due to rhinosinusitis and -0.60 (95% CI, -1.41 to 0.21) in patients with a positive rhinoscopy result. At 7 days patients who took amoxicillin-clavulanate were more likely to have diarrhea (odds ratio, 3.89; 95% CI, 2.09-7.25). CONCLUSIONS: Adult patients in general practice with clinically diagnosed acute rhinosinusitis experience no advantage with antibiotic treatment with amoxicillin-clavulanate and are more likely to experience adverse effects. PMID- 12912715 TI - Increased risk of achilles tendon rupture with quinolone antibacterial use, especially in elderly patients taking oral corticosteroids. AB - BACKGROUND: In several case reports, the occurrence of Achilles tendon rupture has been attributed to the use of quinolones, but the epidemiologic evidence for this association is scanty. METHODS: We conducted a population-based case-control study in the General Practice Research Database in the United Kingdom during the period 1988 through 1998. Cases were defined as all persons who had a first-time recording of an Achilles tendon rupture, and who had at least 18 months of valid history before the index date. As a control group, we randomly sampled 50 000 patients with at least 18 months of valid history who were assigned a random date as index date. RESULTS: We identified 1367 cases that met the inclusion criteria. The adjusted odds ratio (OR) for Achilles tendon rupture was 4.3 (95% confidence interval [CI], 2.4-7.8) for current exposure to quinolones, 2.4 (95% CI, 1.5-3.7) for recent exposure, and 1.4 (95% CI, 0.9-2.1) for past exposure. The OR of Achilles tendon rupture was 6.4 (95% CI, 3.0-13.7) in patients aged 60 to 79 years and 20.4 (95% CI, 4.6-90.1) in patients aged 80 years or older. In persons aged 60 years and older, the OR was 28.4 (95% CI, 7.0-115.3) for current exposure to ofloxacin, while the ORs were 3.6 (95% CI, 1.4-9.1) and 14.2 (95% CI, 1.6 128.6) for ciprofloxacin and norfloxacin, respectively. Approximately 2% to 6% of all Achilles tendon ruptures in people older than 60 years can be attributed to quinolones. CONCLUSIONS: Current exposure to quinolones increased the risk of Achilles tendon rupture. The risk is highest among elderly patients who were concomitantly treated with corticosteroids. PMID- 12912716 TI - Clinicians' responses to direct-to-consumer advertising of prescription medications. AB - BACKGROUND: The direct-to-consumer advertising (DTCA) of prescription medications is proliferating in the United States. The relationship between patient exposure to DTCA and the response of clinicians is not well understood. METHODS: A randomized postal survey of Arizona primary care provider physicians (n = 1080) and physician assistants (n = 704) was conducted. A questionnaire was created using a hypothetical patient scenario that varied according to the diagnosis of the patient (ie, hypertension, hypercholesterolemia, seasonal allergies, or obesity) and the type of informational exposure generating the patient's questions (ie, DTCA vs drug references such as Physicians' Desk Reference). Clinicians were randomly assigned 1 of 8 forms of the scenario and were asked standardized questions related to their responses when faced with the patient scenario. RESULTS: The response rate was 44% (40.5% of physicians and 49.3% of physician assistants). No statistically significant differences were found between the early and late responders or between responders and nonresponders. Relative to clinicians who received the "drug reference book" patient scenario, clinicians who received the DTCA patient scenario were more likely to become annoyed with a patient for asking for more information about medications (P =.003); less likely to answer the patient's questions (P =.03) or provide additional written information (P =.007); more likely to become frustrated (P =.003) and annoyed (P<.001) with the patient for asking to try a specific medication; and less likely to provide samples (P =.001) or a prescription (P<.001) for a specific medication. CONCLUSION: Clinicians are amenable to patients asking for drug information and medications, but they are less receptive to questions arising from DTCA. PMID- 12912717 TI - Quality assessment of a collaborative approach for decreasing drug-related morbidity and achieving therapeutic goals. AB - BACKGROUND: Collaboration between physicians and pharmacists is one approach to address drug-related morbidity and achieve therapeutic goals. A collaborative practice of pharmaceutical care has been used in the Fairview Clinics System of Minneapolis-St Paul since 1999. METHODS: The quality of therapeutic determinations made by pharmacists within this collaborative practice of pharmaceutical care was studied by a 12-member panel of physicians and pharmacists who used randomly selected patient records. This was a quality improvement and care process validation component of a study evaluating the effects of drug therapy management in patients receiving prepaid medical assistance. An implicit review process was used to evaluate the clinical credibility of therapeutic determinations made by pharmaceutical care practitioners. RESULTS: A total of 5780 drug therapy problems were resolved for 2524 patients receiving pharmaceutical care. The rate of therapeutic goals achieved increased from 74% at the time of patients' initial pharmaceutical care encounters to 89% at patients' latest encounters. In this quality assessment analysis panel members performed a total of 4779 evaluations of clinical decisions. Panelists indicated agreement with the evaluations in 94.2% of cases, expressed a neutral opinion in 3.6% of cases, and disagreed in 2.2% of cases. Intraclass correlation coefficients ranged from 0.73 to 0.85. CONCLUSIONS: The decisions made by pharmaceutical care practitioners working in collaboration with physicians to provide drug therapy management services are clinically credible based on the evaluations and comments of a peer review panel. This study provides information on the quality of care provided by pharmacists when collaborating with physicians to provide drug therapy management services. PMID- 12912718 TI - Increased prevalence of aortic stenosis in patients with arteriovenous malformations of the gastrointestinal tract in Heyde syndrome. AB - BACKGROUND: Heyde syndrome is described as the association of arteriovenous malformations (AVMs) of the gastrointestinal tract and aortic stenosis (AS); its existence, however, has been questioned. We examined whether there is an association between AVMs and AS when objective measures are used to diagnose these findings. METHODS: We identified all patients who were diagnosed with AVMs between 1990 and 2000 by means of gastrointestinal endoscopy or mesenteric angiography. We compared the prevalence of AS and mitral stenosis (MS) in the 73 patients with AVMs who also had echocardiograms. For a comparison with the general population, the prevalence of AS and MS in all patients who had echocardiograms between 1990 and 2000 was calculated (MS was chosen for comparison as a lesion with similar likelihood of prompting an echocardiographic evaluation). RESULTS: The prevalence of AS was 31.7% in patients with AVMs, which was significantly higher than the 14.0% found in the general population comparison group (P<.001). The prevalence of MS was 1.6% in the AVM group, which was not statistically different from the 6.0% MS prevalence in the general echocardiogram population (P =.14). Significant AS was 2.6 times more common, and severe AS was 4.1 times more common, in patients with AVMs than in the general population. Age and sex were not associated with Heyde syndrome, but the association was more prevalent in blacks. CONCLUSIONS: Our study confirmed an association between AVMs and AS, although the etiology of the Heyde syndrome remains unclear. Clinicians need to be aware of this syndrome because it may affect their management of patients with gastrointestinal bleeding or AS. PMID- 12912720 TI - Symptoms of rhinosinusitis in patients with unexplained chronic fatigue or bodily pain: a pilot study. AB - BACKGROUND: Recent otolaryngologic studies document significant fatigue and bodily pain (BP) in patients with chronic rhinosinusitis. Studies of general medical patients are lacking. METHODS: A case-control study of 297 consecutive general medical outpatients. RESULTS: Sixty-five patients noted unexplained chronic fatigue (UCF), 33 reported BP, and 26 had both. Compared with 232 patients without UCF, patients with UCF more frequently had the following rhinosinusitis symptoms: facial pressure (odds ratio [OR], 9.7; 95% confidence interval [CI], 5.2-18.2), heavy-headedness (OR, 21.9; 95% CI, 10.9-44.0), nasal obstruction (OR, 4.3; 95% CI, 2.3-7.9), frontal headache (OR, 13.6; 95% CI, 6.5 28.5), postnasal drip (OR, 2.8; 95% CI, 1.6-5.0), sore throat (OR, 3.1; 95% CI, 1.5-6.6), and tender cervical lymph nodes (OR, 9.2; 95% CI, 4.3-19.7). A similar predominance of rhinosinusitis symptoms was noted in patients with BP and in 15 patients with UCF who had chronic fatigue syndrome. No increased prevalence of pollen allergy was noted in association with UCF, BP, or chronic fatigue syndrome. Gastrointestinal, sleep, and psychiatric problems were similar between patients with UCF and 38 patients with explained fatigue. Rhinosinusitis symptoms, however, were more common in UCF. CONCLUSIONS: There is an increased prevalence of rhinosinusitis symptoms but not pollen allergy among general medical outpatients with UCF, BP, or both. Rhinosinusitis symptoms are at least as common as gastrointestinal complaints, sleep disturbance, and psychiatric problems (previously well documented complaints associated with UCF and BP). Rhinosinusitis symptoms, furthermore, are more common in UCF than in fatigue explained by a physical or mental illness. PMID- 12912719 TI - Hypoglycemia as a predictor of mortality in hospitalized elderly patients. AB - BACKGROUND: Hypoglycemia during hospitalization occurs in patients with and without diabetes. The aims of this study were to determine the incidence, associated risk factors, and short- and long-term outcome of hypoglycemia among hospitalized elderly patients. METHODS: This is a case-control study conducted at geriatric and medicine departments. All patients 70 years or older with documented hypoglycemia hospitalized within 1 year (n = 281) were compared with a nonhypoglycemic group of 281 elderly, randomly selected patients from the same hospitalized population. RESULTS: Among 5404 patients 70 years or older, 281 (5.2%) had documented hypoglycemia. Compared with the nonhypoglycemic group, we found the following characteristics to be true in the hypoglycemic group: there were more women than men (58% vs 44%, P =.001); sepsis was 10 times more common (P<.001); malignancy was 2.8 times more common (P =.04); the mean serum albumin level was lower (2.8 g/dL vs 3.4 g/dL, P<.001); and the mean serum creatinine and alkaline phosphatase levels were higher (P<.001 for both). Diabetes was known in 42% of the hypoglycemic group and in 31% of the nonhypoglycemic group (P =.03); 70 patients in the hypoglycemic group were taking sulfonylureas or insulin. Multivariate logistic analysis showed that sepsis, albumin level, malignancy, sulfonyurea and insulin treatment, alkaline phosphatase level, female sex, and creatinine level were all independent predictors of developing hypoglycemia. In hospital mortality and 3-month mortality were about twice as high in the hypoglycemic group (P<.001). Multivariate analysis of mortality found that sepsis, low albumin level, and malignancy were independent predictors, while hypoglycmia was not. CONCLUSIONS: Hypoglycemia was common in elderly hospitalized patients and predicted increased in-hospital 3- and 6-month cumulative mortality. However, in a multivariate analysis, hypoglycemia was not an independent predictor for mortality, implying that it is only a marker. PMID- 12912722 TI - Venous thromboembolic disease: comparison of the diagnostic process in blacks and whites. AB - BACKGROUND: There has been concern that a disproportionate use of some health services exists among races. Whether this applies to patients with pulmonary embolism (PE) or deep venous thrombosis (DVT) has not been determined. OBJECTIVE: To assess if there is a racial disparity in the application of diagnostic tests for PE or DVT, or in reaching a diagnosis or using medical facilities. DESIGN: A study of cross-sectional samples of hospitalizations during 21 years using data from the National Hospital Discharge Survey. SETTING: Noninstitutional hospitals in 50 states and the District of Columbia from January 1, 1979, through December 31, 1999. PATIENTS: The National Hospital Discharge Survey abstracts demographic and medical information from the medical records of inpatients. For 1979 through 1999, the number of patients sampled ranged annually from 181 000 to 307 000. Measurements The number of sampled patients with DVT and with PE and the number of diagnostic tests performed were determined from the International Classification of Diseases, Ninth Revision, Clinical Modification codes at discharge. A multistage estimation procedure gave an estimate of values for the entire United States. RESULTS: The age-adjusted rates of diagnosis of PE and of DVT per 100 000 population were not lower in blacks than in whites. Rates of use of radioisotopic lung scans, venous ultrasonography of the lower extremities, and contrast venography were comparable between races. The durations of hospitalization for patients with a primary discharge diagnosis of PE and of DVT were also comparable. CONCLUSIONS: There is nothing to suggest that diagnostic tests are being withheld, and there is no evidence of a failure to reach a diagnosis in blacks with thromboembolic disease. PMID- 12912721 TI - Regression of carotid and femoral artery intima-media thickness in familial hypercholesterolemia: treatment with simvastatin. AB - OBJECTIVE: To investigate whether high-dose simvastatin therapy could reduce carotid and femoral artery intima-media thickness (IMT) in patients with familial hypercholesterolemia (FH) to prevent cardiovascular disease. BACKGROUND: Imaging of arterial walls with B-mode ultrasonography is increasingly used as a noninvasive surrogate marker of cardiovascular disease. Intervention trials using this modality have shown that by reducing risk factors, progression of atherosclerosis was inhibited. METHODS: After a washout period of 6 weeks, all patients with FH started monotherapy with simvastatin, 80 mg/d, for 2 years. The primary end point was the change (in millimeters) of the mean combined far-wall IMT of predefined carotid and femoral arterial segments at 2 years. RESULTS: We included a total of 153 patients with FH. Mean +/- SD combined baseline IMT was 1.07 +/- 0.23 mm. After treatment with simvastatin for 2 years, this IMT decreased by a mean of 0.081 mm (95% confidence interval, -0.109 to -0.053; P<.001), with its largest reduction in the femoral artery (-0.283 mm; P<.001). An actual decrease of combined IMT was seen in 69.8% of all patients. CONCLUSIONS: High-dose simvastatin therapy reduces arterial wall IMT in more than two thirds of the patients, with its largest effect on the femoral artery. Furthermore, patients with FH who were treated with both statin and antihypertensive medication experienced a significantly greater benefit in terms of IMT reduction. PMID- 12912723 TI - Argatroban anticoagulation in patients with heparin-induced thrombocytopenia. AB - BACKGROUND: Heparin-induced thrombocytopenia (HIT) is an intensely prothrombotic syndrome managed by discontinuation of heparin therapy and substitution of an alternative inhibitor of thrombin. We describe our experience with argatroban, a direct thrombin inhibitor, in patients with HIT or HIT with thrombosis (HITTS). METHODS: In this multicenter, nonrandomized prospective study, 418 patients with HIT were administered intravenous argatroban, 2 micro g/kg per minute, adjusted to maintain the activated partial thromboplastin time at 1.5 to 3 times the baseline value for a mean of 5 to 7 days. Comparisons were made with a historical control cohort (n = 185). The prospectively defined, primary efficacy end point was a composite of all-cause death, all-cause amputation, or new thrombosis in 37 days. Other end points included the components of the composite, death due to thrombosis, increased platelet count, and bleeding. RESULTS: In the HIT arm, the composite end point was significantly reduced in argatroban-treated patients vs controls (28.0% vs 38.8%; P =.04). In the HITTS arm, the composite end point occurred in 41.5% of argatroban-treated patients vs 56.5% of controls (P =.07). By time-to-event analysis of the composite end point, argatroban therapy was significantly better than historical control therapy in HIT (P =.02) and HITTS (P =.008). Argatroban therapy also significantly reduced new thrombosis in HIT and HITTS and death due to thrombosis in HITTS. There were no significant between group differences in all-cause death or amputation. Platelet counts recovered more rapidly in argatroban-treated patients than in controls. Bleeding rates were similar between groups. CONCLUSION: Argatroban therapy, compared with historical control, improves outcomes, particularly new thrombosis and death due to thrombosis, in patients with heparin-induced thrombocytopenia. PMID- 12912724 TI - Two "HIV-infected" persons not really infected. PMID- 12912725 TI - Food allergy: preventing a fatal outcome. PMID- 12912726 TI - Food allergens and blood transfusions: a cause for concern? PMID- 12912727 TI - What now about acetaminophen? PMID- 12912728 TI - Interpretation of therapies for knee arthritis. PMID- 12912729 TI - Limitation to exercise tolerance in chronic obstructive pulmonary disease: look to the muscles of ambulation. PMID- 12912730 TI - Genetic factors in idiopathic pulmonary fibrosis: transforming growth factor-beta implicated at last. PMID- 12912731 TI - Isoniazid for latent tuberculosis infection: approaching 40 and reaching its prime. PMID- 12912732 TI - Meconium aspiration syndrome: is surfactant lavage the answer? PMID- 12912733 TI - "What we've got here is a failure to communicate". PMID- 12912734 TI - Management of severe acute respiratory syndrome: the Hong Kong University experience. PMID- 12912735 TI - Identifying smokers at risk of COPD and GOLD. PMID- 12912736 TI - The molecular-clinical divorce. PMID- 12912737 TI - Glucocorticoids in preterm infants and discrepancies of vascular endothelial growth factor. PMID- 12912738 TI - San Francisco General Hospital. PMID- 12912739 TI - What is the future of surgery? PMID- 12912740 TI - T1a breast carcinoma and the role of axillary dissection. AB - HYPOTHESIS: Axillary dissection (AD) does not affect recurrence or survival in T1a breast cancer. DESIGN: Cohort study comparing patients who underwent AD and those who did not. SETTING: Provincial cancer agency. PATIENTS: Six hundred ninety-one women with pathologically diagnosed T1a tumors. MAIN OUTCOME MEASURES: Rates of axillary metastases stratified according to grade and lymphovascular and/or neural invasion, rates of relapse, and disease-specific survival. RESULTS: Grade 1, 2, and 3 tumors without lymphovascular and/or neural invasion had axillary nodal involvement rates of 0.7%, 7%, and 7.8% of patients, respectively; with lymphovascular and/or neural invasion, axillary nodes were involved in 9.1%, 39.3%, and 44.4%, respectively. No statistically significant differences were found between the cohorts in relapse rates (P =.70) or survival (P =.84). CONCLUSION: Higher tumor grade and lymphovascular and/or neural invasion increased the rate of nodal metastases in T1a tumors, but AD did not improve relapse rates or breast cancer-specific survival. PMID- 12912741 TI - Enhanced trauma program commitment at a level I trauma center: effect on the process and outcome of care. AB - HYPOTHESIS: With advances in surgical care, the occurrences of major adverse outcomes have become a rare event. The effect of a surgical service can be more comprehensively evaluated by following the Donabedian model, looking at the triad of structure, process, and outcome. It is hypothesized that the implementation of a focused program commitment at a trauma center is associated with improvements in process of care and patient outcomes. DESIGN: Evaluation of prospectively collected information in a trauma registry for the 3-year periods immediately before (1995-1997) and after (1999-2001) the implementation (in 1998) of the full time trauma service. SETTING: Level I university-affiliated trauma center. PATIENTS: Patients meeting criteria for major trauma. INTERVENTION: The implementation of a full-time trauma service, featuring 24-hour in-house attending coverage, dedicated trauma admitting unit, regular trauma core curriculum, regular multidisciplinary quality assurance meetings, and state designation for level I status. MAIN OUTCOME MEASURES: Process of care measures, including time in the emergency department (ED) and trauma "bypass" hours (ie, time spent in the trauma resuscitation area). Outcome measures, including lengths of stay, overall mortality and mortality, excluding ED deaths. RESULTS: The total number of patients with major trauma increased from 2240 (1995-1997) to 2513 (1999-2001). The average time in the ED for patients going to the operating room, intensive care unit, and observation wards all decreased significantly (84 vs 52 minutes, 197 vs 118 minutes, and 300 vs 140 minutes, respectively; all with P<.01). The number of hours that the trauma center was closed owing to ED overcrowding also decreased significantly, from 56 to 2.7 hours per month (P<.01). After excluding ED deaths, there was a trend on bivariate analyses toward lower overall mortality rates (4.5% vs 3.4%, P =.07) and mortality rates among patients with severe head injury (23.8% vs 17.2%, P =.07). On further analyses with multiple logistic regression, controlling for age, Injury Severity Score, Abbreviated Injury Score (for a head injury), and admission blood pressure, the later period is associated with a 31% decrease in overall odds of death (P =.047) and a 42% decrease in odds of death among patients with severe head injury (an Abbreviated Injury Score, >or=3; P =.03). CONCLUSION: The implementation of a full-time trauma service is associated with improved timeliness of triage and therapeutic interventions and improved patient outcomes. PMID- 12912742 TI - Nonoperative treatment of blunt injury to solid abdominal organs: a prospective study. AB - HYPOTHESIS: Nonoperative management (NOM) of injuries to the liver, spleen, and kidney is highly successful, as shown in retrospective studies, but needs prospective validation. Patients in whom NOM is likely to fail can be identified by specific criteria. DESIGN: Prospective observational study. SETTING: Academic level I trauma center at a county hospital. PATIENTS: Two hundred six patients with injuries to the liver (n = 99), spleen (n = 103), and/or kidney (n = 40). MAIN OUTCOME MEASURES: Failure of NOM. RESULTS: Fifty-seven patients (28%) underwent immediate operation; among the other 149, NOM failed in 33 (22%). The rate of failure for spleen injury (34%) was higher than for liver (17%) or kidney injury (18%) (P<.01). Failure of NOM was due to delayed bleeding from a solid viscus in 20 of the 33 patients. Intestinal injury was detected in only 1 patient initially selected for NOM. Specifically among patients with liver injury, no failure was due to delayed bleeding from the liver. Patients with failed NOM were more likely to have a positive abdominal ultrasonographic finding (61% vs 22%; P<.01), a grade of splenic injury of at least III on computed tomographic scan (CT) (n = 20 [17%] vs n = 16 [48%]; P<.01), and an amount of free fluid of greater than 300 mL on CT (36% vs 8%; P<.01) and to receive blood transfusions during NOM (58% vs 16%; P<.01). The groups were not different with regard to associated extra-abdominal injuries (including head injuries). Mortality was not different, but morbidity was marginally higher in patients with failed NOM (29% vs 45%; P =.08). We identified the following 4 independent risk factors of failure by means of stepwise logistic regression: nonliver (splenic or renal) injury, positive abdominal ultrasonography findings, amount of free fluid on CT of greater than 300 mL, and need for blood transfusion. According to a statistical model, the presence of all 4 independent risk factors predicted NOM failure in 96% of the patients, and the absence of all predicted success in 98%. CONCLUSIONS: In a prospective study, the rate of NOM failure for solid abdominal organ injuries is higher than the rates reported in retrospective studies. Nonoperative management is less likely to fail in liver injuries than in splenic or kidney injuries. Use of NOM should be exercised with caution if blood transfusion is needed, fluid is identified on the screening ultrasonogram, or a significant quantity of blood is discovered on CT. PMID- 12912743 TI - High prevalence of potentially hepatotoxic herbal supplement use in patients with fulminant hepatic failure. AB - HYPOTHESIS: The use of potentially hepatotoxic herbal and dietary supplements is highly prevalent in the fulminant hepatic failure (FHF) patient population at our institution, and this subgroup of patients has a worse prognosis. DESIGN: Retrospective case series. Settings An adult tertiary care university hospital and a Veterans Affairs hospital in Oregon. PATIENTS: All patients referred to the liver transplantation service for FHF from January 2001 through October 2002 (N = 20). We defined FHF as onset of encephalopathy within 8 weeks of onset of jaundice in the absence of preexisting liver disease. All patients underwent investigation for potential causes of liver injury. Potentially hepatotoxic supplements were defined as those with previously published reports of hepatic injury related to their use. RESULTS: Ten patients (50%) were recent or active users of potentially hepatotoxic supplements or herbs; 10 had no history of supplement use. In the supplement group, 7 patients (35%) had no other identified cause for hepatic failure. Six patients in the supplement group and 2 patients in the nonsupplement group underwent orthotopic liver transplantation. Five patients in each group died. There were no significant differences in transplantation rate (P =.07) or survival (P>.99) between groups. Supplement use alone accounted for the most cases of FHF during this period, exceeding acetaminophen toxicity and viral hepatitis. CONCLUSIONS: Herbal and dietary supplements were potential hepatotoxins in a high proportion of patients with FHF at our institution. Enhanced public awareness of the potential hepatotoxicity of these commonly used agents and increased regulatory oversight of their use is strongly urged. PMID- 12912744 TI - Morbidity and mortality of aggressive resection in patients with advanced neuroendocrine tumors. AB - BACKGROUND: There is considerable controversy about the treatment of patients with malignant advanced neuroendocrine tumors of the pancreas and duodenum. Aggressive surgery remains a potentially efficacious antitumor therapy but is rarely performed because of its possible morbidity and mortality. HYPOTHESIS: Aggressive resection of advanced neuroendocrine tumors can be performed with acceptable morbidity and mortality rates and may lead to extended survival. DESIGN: The medical records of patients with advanced neuroendocrine tumors who underwent surgery between 1997 and 2002 by a single surgeon at the University of California, San Francisco, were reviewed in an institutional review board approved protocol. MAIN OUTCOME MEASURES: Surgical procedure, pathologic characteristics, complications, mortality rates, and disease-free and overall survival rates were recorded. Disease-free survival was defined as no tumor identified on radiological imaging studies and no detectable abnormal hormone levels. Proportions were compared statistically using the Fisher exact test. Kaplan-Meier curves were used to estimate survival rates. RESULTS: Twenty patients were identified (11 men and 9 women). Of these, 10 (50%) had gastrinoma, 1 had insulinoma, and the remainder had nonfunctional tumors; 2 had multiple endocrine neoplasia type 1, and 1 had von Hippel-Lindau disease. The mean age was 55 years (range, 34-72 years). In 10 patients (50%), tumors were thought to be unresectable according to radiological imaging studies because of multiple bilobar liver metastases (n = 6), superior mesenteric vein invasion (n = 3), and extensive nodal metastases (n = 1). Tumors were completely removed in 15 patients (75%). Surgical procedures included 8 proximal pancreatectomies (pancreatoduodenectomy or whipple procedure), 3 total pancreatectomies, 9 distal pancreatectomies, and 3 tumor enucleations from the pancreatic head. Superior mesenteric vein reconstruction was done in 3 patients. Liver resections were done in 6 patients, and an extended periaortic node dissection was performed in 1. The spleen was removed in 11 patients, and the left kidney was removed as a result of tumor metastases in 2. Eighteen patients had primary pancreatic tumors, and 2 had duodenal tumors; 2 patients with multiple endocrine neoplasia type 1 had both pancreatic and duodenal tumors. The mean tumor size was 8 cm (range, 0.5-23 cm). Of the patients, 14 (70%) had lymph node metastases and 8 (40%) had liver metastases. The mean postoperative hospital stay was 11.5 days (range, 6-26 days). Six patients (30%) had postoperative complications. There was a significantly greater incidence of pancreatic fistulas with enucleations compared with resections (P =.04). There were no operative deaths. The mean follow-up period was 19 months (range, 1-96 months); 18 patients (90%) are alive, 2 died of progressive tumor, and 12 (60%) are disease-free. The actuarial overall survival rate is 80% at 5 years, and disease-free survival rates indicate that all tumors will recur by the 7-year follow-up visit. CONCLUSIONS: Aggressive surgery including pancreatectomy, splenectomy, superior mesenteric vein reconstruction, and liver resection can be done with acceptable morbidity and low mortality rates for patients with advanced neuroendocrine tumors. Although survival rates following surgery are excellent, most patients will develop a recurrent tumor. These findings suggest that conventional contraindications to surgical resection, such as superior mesenteric vein invasion and nodal or distant metastases, should be reconsidered in patients with advanced neuroendocrine tumors. PMID- 12912745 TI - Parathyroidectomy for primary hyperparathyroidism in octogenarians and nonagenarians: a plea for early surgical referral. AB - BACKGROUND: Surgical referral of elderly patients with PHPT is often deferred because of the perceived risks of general anesthesia, increased age, and comorbidities and also because many of the symptoms of PHPT are difficult to distinguish from the aging process. HYPOTHESIS: Parathyroidectomy for primary hyperparathyroidism (PHPT) in elderly patients is beneficial and safe. DESIGN: Retrospective study of patients with PHPT who were 80 years and older. SETTING: Tertiary academic referral medical center. PATIENTS: Fifty-four consecutive patients (43 women and 11 men) with a mean age of 83.6 years who underwent parathyroidectomy for PHPT. MAIN OUTCOME MEASURES: Delay in surgical referral; improvement in symptoms and normalization of serum calcium and parathyroid hormone levels; and perioperative morbidity and mortality. RESULTS: Twelve (22%) of 54 patients had a mean delay of 5 years (range, 1.2-10.0 years) before surgical referral for a known diagnosis of PHPT. No patient had asymptomatic PHPT. There was no significant difference in the preoperative mean serum calcium level and symptoms of PHPT resulting from delay in surgical referral. Parathyroidectomy resulted in significant improvement of PHPT-associated symptoms of fatigue, weight loss, nocturia, bone pain, constipation, and major depression (P<.05). All patients had normal serum calcium and parathyroid hormone levels postoperatively. The average length of hospitalization was 1.9 days. There were 3 complications (9%): 1 urinary tract infection, 1 bladder perforation, and 3 transient hypocalcemia. There was no perioperative mortality. CONCLUSIONS: Parathyroidectomy in elderly patients is safe, curative, and beneficial. Surgical referral for parathyroidectomy should be considered in elderly patients with PHPT. PMID- 12912746 TI - In utero repair of myelomeningocele: experimental pathophysiology, initial clinical experience, and outcomes. AB - HYPOTHESIS: Experimental work raises the possibility that in utero repair of myelomeningocele (MMC) may improve lower extremity, bladder, and bowel function, ameliorate the Arnold-Chiari malformation, and decrease the need for postnatal shunting. DESIGN: We previously developed fetal lamb models to create and reverse lower extremity damage and the Arnold-Chiari malformation in utero. We then applied our extensive experience with fetal surgery, including fetal endoscopic (fetoscopic) surgical manipulation, to develop techniques for MMC repair. SETTING: A tertiary referral center. PATIENTS: All patients treated between 1998 and 2002 for a prenatally diagnosed MMC. INTERVENTIONS: Either fetoscopic MMC repair, fetoscopic patch repair, or limited maternal hysterotomy and microsurgical 3-layered fetal MMC repair was performed. MAIN OUTCOME MEASURES: Gestational age at delivery, survival, neurologic outcome, and need for ventricular shunting at 1 year. RESULTS: Complete fetoscopic repair was accomplished in 1 fetus. Two other fetuses underwent partial fetoscopic procedures. The remaining 10 patients underwent limited maternal hysterotomy and microsurgical 3-layered fetal MMC repair. Four of 13 patients died, and the mean gestational age at delivery of 11 fetuses born alive was 31 weeks. Five of 9 required ventricular shunting by age 1 year. In 2 patients, lower extremity function improved by more than 2 vertebral levels compared with prenatal ultrasonography. Five of 10 patients who lived longer than 3 weeks required postnatal wound revision within 7 days after birth. CONCLUSIONS: Fetoscopic repair, although feasible, does not yet yield optimal surgical results. Open surgical repair before 22 weeks' gestation is physiologically sound and technically feasible. One third of patients appear to be spared the need for a shunt at age 1 year, but improvement in distal neurologic function is less clear. Additionally, fetal mortality is associated with this procedure. Our results complement the data published by groups at Children's Hospital of Philadelphia, in Pennsylvania, and Vanderbilt University, Nashville, Tenn. A National Institutes of Health-sponsored prospective randomized trial is now underway at these 3 centers to compare fetal repair with postnatal repair. PMID- 12912747 TI - Dehydroepiandrosterone sulfate causes proliferation of estrogen receptor-positive breast cancer cells despite treatment with fulvestrant. AB - HYPOTHESIS: Dehydroepiandrosterone sulfate (DHEA-S) causes a proliferation of estrogen receptor (ER)-positive breast cancer cells, even with tamoxifen citrate blockade. The ER antagonist ICI 182780 (fulvestrant) will more effectively stop the proliferative effect of DHEA-S on breast cancer cells. DESIGN: Examination of in vitro breast cancer cell growth in the presence of fulvestrant and DHEA-S. SETTING: Surgical oncology research laboratory. INTERVENTIONS: The ER-positive and ER-negative breast cancer cells were pretreated with fulvestrant and stimulated with 900 microg/dL (22.8 micromol/L) of DHEA-S. MAIN OUTCOME MEASURES: Assays using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, thiazolyl blue, were performed on the third, fifth, and seventh days poststimulation and permitted the calculation of growth percent change. RESULTS: The ER-positive and progesterone receptor-positive cells demonstrated universal proliferation of 107% by day 7 when treated with fulvestrant, regardless of the dose. The ER-negative and progesterone receptor-negative cells demonstrated growth inhibition. CONCLUSIONS: The DHEA-S circumvented fulvestrant inhibition and caused ER-positive breast cancer cell growth. PMID- 12912748 TI - Surgical outcomes of a breast cancer-screening program for low-income women. AB - HYPOTHESIS: Surgical outcomes from a breast cancer-screening program of low income women are similar to those of other screening programs. DESIGN: Prospective cohort. SETTING: Federally funded screening program. PATIENTS: A total of 15730 women. INTERVENTIONS: A total of 23149 mammograms, 20396 with concomitant clinical breast examination, from January 1, 1997, through December 31, 2001. OUTCOME MEASURES: American College of Radiology scores; associated surgery consultations, biopsies, operations, and pathology results. RESULTS: Most (20868) of the 21296 mammograms assigned an American College of Radiology score were benign; only 428 (2%) were suspicious. Resulting from suspicious clinical breast examinations, the group with American College of Radiology scores of 1 to 3 accounted for 45%, 18%, and 10% of recommended surgical consultations, biopsies, and cancers detected, respectively. A rate of 12.3 cancers per 1000 women was found, greater than with other screening programs. Compliance with therapy was 97%. CONCLUSIONS: This screening program had a higher rate of advanced cancers. Clinical breast examination was an important component, and compliance with surgical recommendations was excellent. PMID- 12912749 TI - The significance of elevated levels of parathyroid hormone in patients with morbid obesity before and after bariatric surgery. AB - HYPOTHESIS: The risk of hyperparathyroidism after the duodenal switch operation is related to the length of the common channel. DESIGN: A retrospective analysis of patients following the duodenal switch operation from October 2, 2000, through February 1, 2002. SETTING: Academic tertiary referral hospital. PATIENTS: One hundred sixty-five consecutive patients underwent the duodenal switch operation, performed for morbid obesity, with common channel lengths of 75 cm (n = 103 [group A]) and 100 cm (n = 62 [group B]). MAIN OUTCOME MEASURES: Weight loss and parathyroid hormone, corrected calcium, and 25-hydroxyvitamin D (25-OH D) levels were compared between groups A and B. Values were determined preoperatively, early postoperatively (3-6 months), and late postoperatively (9-18 months). RESULTS: Both groups exhibited a slight reduction in serum calcium concentration, with one quarter decreasing below the normal range. Hyperparathyroidism was more common in group A than group B preoperatively (38.9% vs 14.9%), reflecting the higher body mass index of patients in group A. Hyperparathyroidism was also more frequent in the early (54.9% vs 30.9%) and late (49.4% vs 20.5%) postoperative periods in group A vs group B. New-onset hyperparathyroidism was also more common in group A than group B (42.0% vs 13.3%). After 1 year, subnormal 25-OH D levels were found in 17.0% of the patients in group A and in 10.0% of the patients in group B. Median 25-OH D levels increased in both groups, but tended to be higher in group B. CONCLUSIONS: Patients with shorter common channels had a higher risk of developing hyperparathyroidism. This may be related to limited 25-OH D absorption. PMID- 12912750 TI - Results of 101 ruptured abdominal aortic aneurysm repairs from a single surgical practice. AB - Hypotheses The results of ruptured abdominal aortic aneurysm repairs from a solo community hospital-based practice are comparable to those reported from large university referral medical centers. Patients younger than 70 years, arriving in the emergency department with stable hemodynamics, and undergoing prompt operation have better outcome. DESIGN: A retrospective review from an ongoing vascular surgery registry. SETTING: Two midsized (300-bed) community hospitals. One hundred one consecutive patients with ruptured abdominal aortic aneurysms who were undergoing open surgical repair by a single surgeon (S.S.H.) during a 21 year period were reviewed. MAIN OUTCOME MEASURES: Operative mortality; cardiac, pulmonary, renal, and gastrointestinal complications; and coagulation abnormalities were recorded. Iatrogenic complications and length of hospital stay were noted. Preoperative and intraoperative factors affecting mortality were studied. RESULTS: Fifty-three patients survived ruptured abdominal aortic aneurysm repair (operative mortality, 47.5%). A favorable outcome was observed in patients (1). younger than 70 years, (2). with a hematocrit of more than 35% at presentation, and (3). with emergency department to operating room times of less than 120 minutes. Increasing experience of the surgeon did not result in improved survival. CONCLUSION: The results of ruptured abdominal aortic aneurysm repairs from community-based practice are comparable to those reported from university referral medical centers. PMID- 12912752 TI - A rational approach to cholelithiasis in bariatric surgery: its application to the laparoscopically placed adjustable gastric band. AB - BACKGROUND: Gallstones are more common in the obese population and may be formed during rapid weight loss. A rational approach to the management of the gallbladder should be incorporated into bariatric surgical practice. It has been recommended that patients undergoing Roux-en-Y gastric bypass have routine cholecystectomy regardless of gallstone status. We analyzed the outcomes of a noninterventionist policy on 1000 patients undergoing laparoscopic adjustable gastric banding. HYPOTHESIS: Patients scheduled for adjustable gastric banding should undergo investigation for and treatment of gallbladder disease regardless of symptoms. METHODS: Patients were screened preoperatively for symptoms of gallstones. Ultrasound examination was performed only in those with symptoms and, if stones were present, cholecystectomy was performed with gastric banding. The remaining patients were followed up clinically and outcomes were noted. RESULTS: A total of 1000 patients were followed up for 12 to 96 months, a total of approximately 3500 patient-years. Cholecystectomy was performed in 181 patients before and 10 at gastric banding surgery. Of the 809 patients at risk, 55 (6.8%) presented with symptomatic disease during follow-up and proceeded to undergo elective cholecystectomy without complications from the disease or the treatment. CONCLUSIONS: The incidence of cholecystectomy after gastric banding surgery was not different from the expected rate for a nonsurgical obese population. In contrast, after Roux-en-Y gastric bypass, a median of 40% of patients form stones in the postoperative period, and prophylactic cholecystectomy may be justified. Our data indicate that a noninterventionist approach to the gallbladder is appropriate for patients undergoing adjustable gastric banding surgery. PMID- 12912751 TI - Laparoscopic refundoplication with prosthetic hiatal closure for recurrent hiatal hernia after primary failed antireflux surgery. AB - BACKGROUND: One of the most frequent complications after laparoscopic antireflux surgery is estimated to be the intrathoracic herniation of the wrap into the chest. Therefore, in up to 5% of patients, revisional surgery is necessary. HYPOTHESIS: Patients who undergo laparoscopic refundoplication for postoperative intrathoracic wrap herniation using a circular polypropylene mesh for hiatal closure have a good to excellent functional outcome, during a complete follow-up of 1 year. DESIGN: Prospective nonrandomized trial of a consecutive sample. SETTING: University-affiliated community hospital. PATIENTS: Twenty-four patients undergoing laparoscopic refundoplication for persistent or recurrent symptoms of gastroesophageal reflux disease as a result of postoperative intrathoracic wrap migration. INTERVENTION: All patients underwent laparoscopic refundoplication with a circular polypropylene mesh for hiatal closure. MAIN OUTCOME MEASURES: Recurrences, complications, postoperative lower esophageal sphincter pressure, DeMeester score, esophagogastroduodenoscopy results, and barium swallow results. RESULTS: All refundoplications were completed laparoscopically. There were no intraoperative complications. Twenty-one patients underwent laparoscopic Nissen fundoplication; in 3 patients, a laparoscopic Toupet fundoplication was performed. Previous antireflux procedures included an open Nissen fundoplication (n = 5), a laparoscopic Nissen fundoplication (n = 15), and a laparoscopic Toupet fundoplication (n = 4). Postoperatively, one patient had severe dysphagia and had to undergo pneumatic dilatation once. During a follow-up of 1 year after surgery, no patient developed a recurrent hiatal hernia, with or without intrathoracic wrap herniation. The mean lower esophageal sphincter pressure increased significantly (P<.01) at 3 months (12.2 mm Hg) and 1 year (11.9 mm Hg) after refundoplication. The mean DeMeester score decreased significantly (P<.01) from 50.5 points preoperatively to 16.0 points at 3 months and 14.7 points at 1 year after refundoplication. CONCLUSION: Laparoscopic refundoplication with prosthetic hiatal closure is a safe and effective procedure for preventing recurrent intrathoracic wrap herniation, with good to excellent functional outcome for a complete follow-up of 1 year. PMID- 12912753 TI - Image of the month: Zollinger-Ellison syndrome. PMID- 12912754 TI - The arguments against fellowship training and early specialization in general surgery. PMID- 12912755 TI - A standardized technique for right segmental liver resections. AB - HYPOTHESIS: The indications for segmental liver resections are increasing. This type of procedure can be performed by deep wedge transparenchymal transection or by the intrahepatic approach, reaching the portal pedicle through the hilar plate. We devised a systematized way to perform such an operation. DESIGN: Original surgical technique. PATIENTS AND METHODS: Fourteen consecutive patients (8 men and 6 women; mean age, 55 years) underwent right segmental liver resections between July 1, 2001, and July 31, 2002. Seven patients had liver metastasis, 3 had primary liver cancer, 3 had benign lesions, and 1 had gallbladder cancer. The surgery was performed by making 3 small incisions around the hilar plate. With a standardized method, the right posterior and anterior sheaths were reached by combining these incisions. RESULTS: Right segmental liver resection was feasible with the proposed technique in all patients. Intraoperative blood loss was minimal in all cases, and 11 patients did not require blood transfusion. There was no postoperative death. CONCLUSIONS: This operative procedure standardizes the intrahepatic approach to the right portal pedicle for right segmental resections. It may reduce bleeding at the site of hilar plate incisions and the need for main hepatic pedicle clamping and may facilitate the recognition of right posterior and anterior sheaths, with excellent immediate results. PMID- 12912757 TI - Surgery in Taiwan. AB - Taiwan is an island located southeast of the Asian continent. Surgery in Taiwan has changed rapidly during the past 20 years. Approximately 4500 surgeons (including orthopedic surgeons, neurosurgeons, and urologists) serve a population of 23 million on this 36000-km(2) island. Much progress has been made in the development of basic science surgical research, the use of new technology, and the improvement of the quality of care of our patients. National Health Insurance was launched in 1995 and has had a great impact on the clinical practice of surgery. To attract more medical graduates to the specialty of surgery and enrich the resources of manpower, reasonable reimbursement and a new education program with efficient core content but no prolongation of the training period are needed. PMID- 12912758 TI - Surgery too drastic to treat fibromuscular dysplasia in an elderly patient. PMID- 12912759 TI - The obsession of envy (Monomanie de l'envie). PMID- 12912761 TI - Gray and white matter brain abnormalities in first-episode schizophrenia inferred from magnetization transfer imaging. AB - BACKGROUND: Neuroimaging studies suggest that schizophrenia is associated with gray and possibly white matter changes. It is unclear whether these changes are present at illness onset or which brain structures are selectively affected. New imaging methods such as magnetization transfer imaging may be more sensitive than conventional volumetric imaging to the subtle structural brain changes in schizophrenia. METHODS: High-resolution volumetric T1-weighted images and magnetization transfer images were acquired from 30 patients (29 with first episode schizophrenia and 1 with schizophreniform psychoses) and 30 control subjects. Images were processed using voxel-based morphometry, which allows whole brain analysis. RESULTS: Compared with controls, the magnetization transfer ratio (an index of signal loss derived from magnetization transfer imaging) was reduced bilaterally in the medial prefrontal cortex (right greater than left), insula (left greater than right), and white matter incorporating the fasciculus uncinatus (left greater than right) in the patient group. Analysis of the T1 weighted images did not reveal significant volumetric differences between patients and controls. CONCLUSIONS: Gray and white matter abnormalities are present in schizophrenia at illness onset. The magnetization transfer ratio is sensitive to these abnormalities, which cannot be explained by detectable atrophy in our patient group. PMID- 12912760 TI - Progressive decrease of left Heschl gyrus and planum temporale gray matter volume in first-episode schizophrenia: a longitudinal magnetic resonance imaging study. AB - BACKGROUND: The Heschl gyrus and planum temporale have crucial roles in auditory perception and language processing. Our previous investigation using magnetic resonance imaging (MRI) indicated smaller gray matter volumes bilaterally in the Heschl gyrus and in left planum temporale in patients with first-episode schizophrenia but not in patients with first-episode affective psychosis. We sought to determine whether there are progressive decreases in anatomically defined MRI gray matter volumes of the Heschl gyrus and planum temporale in patients with first-episode schizophrenia and also in patients with first-episode affective psychosis. METHODS: At a private psychiatric hospital, we conducted a prospective high spatial resolution MRI study that included initial scans of 28 patients at their first hospitalization (13 with schizophrenia and 15 with affective psychosis, 13 of whom had a manic psychosis) and 22 healthy control subjects. Follow-up scans occurred, on average, 1.5 years after the initial scan. RESULTS: Patients with first-episode schizophrenia showed significant decreases in gray matter volume over time in the left Heschl gyrus (6.9%) and left planum temporale (7.2%) compared with patients with first-episode affective psychosis or control subjects. CONCLUSIONS: These findings demonstrate a left-biased progressive volume reduction in the Heschl gyrus and planum temporale gray matter in patients with first-episode schizophrenia in contrast to patients with first episode affective psychosis and control subjects. Schizophrenia but not affective psychosis seems to be characterized by a postonset progression of neocortical gray matter volume loss in the left superior temporal gyrus and thus may not be developmentally fixed. PMID- 12912762 TI - Life event dimensions of loss, humiliation, entrapment, and danger in the prediction of onsets of major depression and generalized anxiety. AB - BACKGROUND: Although substantial evidence suggests that stressful life events predispose to the onset of episodes of depression and anxiety, the essential features of these events that are depressogenic and anxiogenic remain uncertain. METHODS: High contextual threat stressful life events, assessed in 98 592 person months from 7322 male and female adult twins ascertained from a population-based registry, were blindly rated on the dimensions of humiliation, entrapment, loss, and danger and their categories. Onsets of pure major depression (MD), pure generalized anxiety syndrome (GAS) (defined as generalized anxiety disorder with a 2-week minimum duration), and mixed MD-GAS episodes were examined using logistic regression. RESULTS: Onsets of pure MD and mixed MD-GAS were predicted by higher ratings of loss and humiliation. Onsets of pure GAS were predicted by higher ratings of loss and danger. High ratings of entrapment predicted only onsets of mixed episodes. The loss categories of death and respondent-initiated separation predicted pure MD but not pure GAS episodes. Events with a combination of humiliation (especially other-initiated separation) and loss were more depressogenic than pure loss events, including death. No sex differences were seen in the prediction of episodes of illness by event categories. CONCLUSIONS: In addition to loss, humiliating events that directly devalue an individual in a core role were strongly linked to risk for depressive episodes. Event dimensions and categories that predispose to pure MD vs pure GAS episodes can be distinguished with moderate specificity. The event dimensions that preceded mixed MD-GAS episodes were largely the sum of those that preceded pure MD and pure GAS episodes. PMID- 12912763 TI - The impact of parental status on the risk of completed suicide. AB - BACKGROUND: Although some studies suggest that parenthood is associated with a reduced suicide risk, the impact of children on parental suicide has rarely been documented. METHODS: This study investigates the impact of parental status on the risk of completed suicide in the context of other risk factors. A nested case control design is used, matching for age, sex, and calendar time. The study is based on 4 Danish longitudinal registers, including 18 611 suicides of individuals aged 18 to 75 years from January 1, 1981, to December 31, 1997, and 372 220 matched control subjects. Information about children and subject's individual background is retrieved and merged. Data are analyzed using conditional logistic regression, yielding odds ratios interpreted as incidence rate ratios. RESULTS: The presence of children is protective against suicide in parents in terms of having children and, to a higher degree, having a young child; these effects exist even when adjusted for marital, socioeconomic, and psychiatric status; and their influences are much stronger in women than in men. At the same time, parents of children with a hospitalized psychiatric disorder and parents of children who have died are at an increased risk for suicide. A child dying during early childhood has a strong effect on suicide in parents, and a suicidal death of a child increases the risk of parental suicide more than a nonsuicidal death. The suicide risk is particularly high in the first month after losing a child. CONCLUSIONS: The impact of children on parental suicide can be protective because of having children. It can also be negative, for example, when losing a child, particularly if the child dies during early childhood; the risk is particularly high during the first month after the loss. PMID- 12912765 TI - Depression with atypical features in the National Comorbidity Survey: classification, description, and consequences. AB - BACKGROUND: Atypical depression has been found to be distinct from other types of depression in terms of psychiatric symptom profile and treatment response. However, debate continues regarding its specific characteristics, impact, and diagnostic criteria. The current study was conducted to increase understanding of atypical depression diagnosed using only the reversed vegetative symptoms of hypersomnia and hyperphagia. METHODS: An atypical depression group (n = 304 [36.4% of the depressed sample; 39.0% when weighted to approximate the national population]) was identified within the US National Comorbidity Survey, which assessed psychiatric disorders among a nationally representative sample using the Composite International Diagnostic Interview. The atypical group was identified based on DSM-III-R criteria for a major depressive episode, in addition to atypical features of hypersomnia and hyperphagia. Comparison groups were those with nonatypical depression (n = 532) and individuals without a psychiatric disorder (n = 4071). RESULTS: Compared with nonatypical depression, atypical depression was associated with a greater percentage of women and an earlier age of onset. The atypical group also reported higher rates of most depressive symptoms, suicidal thoughts and attempts, psychiatric comorbidity (panic disorder, social phobia, and drug dependence), disability and restricted activity days, use of some health care services, paternal depression, and childhood neglect and sexual abuse (P<.05). Compared with people without psychiatric disorders, the atypical group reported higher rates of disability and restricted activity days, use of all mental health care services, parental depression, and childhood abuse (P<.001). CONCLUSIONS: This analysis of a nationally representative US sample suggests that overeating and oversleeping can be used to identify an atypical depression subgroup that is distinct from other depressed patients in terms of demographics, psychiatric comorbidities, and abuse history. Findings also suggest that atypical depression is associated with increased distress, suicidal ideation, and disability compared with nonatypical depression. PMID- 12912764 TI - Altered gene expression of brain-derived neurotrophic factor and receptor tyrosine kinase B in postmortem brain of suicide subjects. AB - BACKGROUND: Suicide is a major public health concern. Although authors of many studies have examined the neurobiological aspects of suicide, the molecular mechanisms associated with suicidal behavior remain unclear. Brain-derived neurotrophic factor (BDNF), one of the most important neurotrophins, after binding with and activating receptor tyrosine kinase B (trk B), is directly involved in many physiological functions in the brain, including cell survival and synaptic plasticity. The present study was performed to examine whether the expression of BDNF and/or trk B isoforms was altered in postmortem brain in subjects who commit suicide (hereafter referred to as suicide subjects) and whether these alterations were associated with specific psychopathologic conditions. METHODS: These studies were performed in prefrontal cortex in Brodmann area 9 and hippocampus obtained in 27 suicide subjects and 21 nonpsychiatric control subjects. Levels of messenger RNA and protein levels of BDNF and trk B were determined with competitive reverse transcriptase-polymerase chain reaction and Western blot technique, respectively. The level of neuron specific enolase messenger RNA as a neuronal marker was also determined in these brain areas. RESULTS: Messenger RNA levels of BDNF and trk B were significantly reduced, independently and as a ratio to neuron-specific enolase, in both prefrontal cortex and hippocampus in suicide subjects, as compared with those in control subjects. These reductions were associated with significant decreases in the protein levels of BDNF and of full-length trk B but not trk B's truncated isoform. These changes were present in all suicide subjects regardless of psychiatric diagnosis and were unrelated to postmortem interval, age, sex, or pH of the brain. CONCLUSIONS: Given the importance of BDNF in mediating physiological functions, including cell survival and synaptic plasticity, our findings of reduced expression of BDNF and trk B in postmortem brain in suicide subjects suggest that these molecules may play an important role in the pathophysiological aspects of suicidal behavior. PMID- 12912766 TI - Gambling urges in pathological gambling: a functional magnetic resonance imaging study. AB - BACKGROUND: Gambling urges in pathological gambling (PG) often immediately precede engagement in self-destructive gambling behavior. An improved understanding of the neural correlates of gambling urges in PG would advance our understanding of the brain mechanisms underlying PG and would help direct research into effective treatments. METHODS: Echoplanar functional magnetic resonance imaging was used to assess brain function during viewing of videotaped scenarios with gambling, happy, or sad content. Participants rated the quality and magnitude of their emotional and motivational responses. RESULTS: Men with PG (n = 10) reported mean +/- SD greater gambling urges after viewing gambling scenarios vs control subjects (n = 11) (5.20 +/- 3.43 vs 0.32 +/- 0.60; chi21,19 = 21.71; P<.001). The groups did not differ significantly in their subjective responses to the happy (P =.56) or sad (P =.81) videotapes. The most pronounced between-group differences in neural activities were observed during the initial period of viewing of the gambling scenarios: PG subjects displayed relatively decreased activity in frontal and orbitofrontal cortex, caudate/basal ganglia, and thalamus compared with controls. Distinct patterns of regional brain activity were observed in specific temporal epochs of videotape viewing. For example, differences localized to the ventral anterior cingulate during the final period of gambling videotape viewing, corresponding to the presentation of the most provocative gambling stimuli. Although group differences in brain activity were observed during viewing of the sad and happy scenarios, they were distinct from those corresponding to the gambling scenarios. CONCLUSIONS: In men with PG, gambling cue presentation elicits gambling urges and leads to a temporally dynamic pattern of brain activity changes in frontal, paralimbic, and limbic brain structures. When viewing gambling cues, PG subjects demonstrate relatively decreased activity in brain regions implicated in impulse regulation compared with controls. PMID- 12912767 TI - Prevalence and development of psychiatric disorders in childhood and adolescence. AB - BACKGROUND: This longitudinal community study assessed the prevalence and development of psychiatric disorders from age 9 through 16 years and examined homotypic and heterotypic continuity. METHODS: A representative population sample of 1420 children aged 9 to 13 years at intake were assessed annually for DSM-IV disorders until age 16 years. RESULTS: Although 3-month prevalence of any disorder averaged 13.3% (95% confidence interval [CI], 11.7%-15.0%), during the study period 36.7% of participants (31% of girls and 42% of boys) had at least 1 psychiatric disorder. Some disorders (social anxiety, panic, depression, and substance abuse) increased in prevalence, whereas others, including separation anxiety disorder and attention-deficit/hyperactivity disorder (ADHD), decreased. Lagged analyses showed that children with a history of psychiatric disorder were 3 times more likely than those with no previous disorder to have a diagnosis at any subsequent wave (odds ratio, 3.7; 95% CI, 2.9-4.9; P<.001). Risk from a previous diagnosis was high among both girls and boys, but it was significantly higher among girls. Continuity of the same disorder (homotypic) was significant for all disorders except specific phobias. Continuity from one diagnosis to another (heterotypic) was significant from depression to anxiety and anxiety to depression, from ADHD to oppositional defiant disorder, and from anxiety and conduct disorder to substance abuse. Almost all the heterotypic continuity was seen in girls. CONCLUSIONS: The risk of having at least 1 psychiatric disorder by age 16 years is much higher than point estimates would suggest. Concurrent comorbidity and homotypic and heterotypic continuity are more marked in girls than in boys. PMID- 12912768 TI - Neurotoxicity, neuroplasticity, and magnetic resonance imaging morphometry. PMID- 12912769 TI - Posttraumatic stress disorder among criminally involved youth. PMID- 12912770 TI - Relapse in bulimia nervosa. PMID- 12912771 TI - Standardized mortality ratio in bulimia nervosa. PMID- 12912772 TI - Influencing the childhood behaviors that lead to obesity: role of the pediatrician and health care professional. PMID- 12912773 TI - Development of novel medications for use in the treatment of obesity in children will be directed by delineating controls of energy homeostasis. PMID- 12912774 TI - Pediatric obesity policy: the danger of skepticism. PMID- 12912775 TI - Obesity prevention in pediatric primary care: four behaviors to target. PMID- 12912776 TI - Inactivity and inaction: we can't afford either. PMID- 12912777 TI - Associations of weight-based teasing and emotional well-being among adolescents. AB - BACKGROUND: Verbal harassment, such as bullying and hate speech, has received considerable attention recently, but less is known about weight-based teasing and its potential harmful effects on young people's psychosocial well-being. OBJECTIVE: To determine the associations of weight-based teasing and body satisfaction, self-esteem, depressive symptoms, and suicidal ideation and suicide attempts using a large sample of adolescents. DESIGN: Secondary analysis of survey and anthropometric data. SETTING: Ethnically and socioeconomically diverse communities in the urban and suburban school districts of the Minneapolis/St Paul metropolitan area. PARTICIPANTS: A school-based sample of 4746 adolescents in grades 7 to 12 at 31 public middle schools and high schools. MAIN OUTCOME MEASURES: Weight-based teasing from peers or family members, body satisfaction, self-esteem, depressive symptoms, and suicidal ideation and suicide attempts. RESULTS: Of the eligible students, 81.5% participated; 30.0% of adolescent girls and 24.7% of adolescent boys were teased by peers, and 28.7% of adolescent girls and 16.1% of adolescent boys were teased by family members. Approximately 14.6% of adolescent girls and 9.6% of adolescent boys reported teasing from both of these sources. Teasing about body weight was consistently associated with low body satisfaction, low self-esteem, high depressive symptoms, and thinking about and attempting suicide, even after controlling for actual body weight. These associations held for adolescent boys and girls, across racial, ethnic, and weight groups. Furthermore, teasing from 2 sources was associated with a higher prevalence of emotional health problems than either teasing from a single source or no teasing. CONCLUSIONS: Physicians and other health care providers should recognize the importance of weight-based teasing for young patients. Policy, programs, and education should focus on increasing awareness of what constitutes weight-based teasing, its potentially harmful effects on adolescents' emotional well-being, and reduction of this behavior. PMID- 12912778 TI - A longitudinal evaluation of adolescent depression and adult obesity. AB - BACKGROUND: Prior studies have had conflicting results regarding the relationship between adolescent depression and adult obesity. OBJECTIVE: To test the hypothesis that depression in adolescence would increase the risk for obesity in early adulthood. METHODS: We used data from a longitudinal study of a birth cohort of children born between April 1, 1972, and March 31, 1973, in Dunedin, New Zealand (N = 1037). These data included regular diagnostic mental health interviews and height/weight measurements throughout childhood and adolescence. We performed logistic regression analyses to assess the relationship between major depression in early or late adolescence and the risk for obesity at 26 years of age. RESULTS: Major depression occurred in 7% of the cohort during early adolescence (11, 13, and 15 years of age) and 27% during late adolescence (18 and 21 years of age). At 26 years of age, 12% of study members were obese. After adjusting for each individual's baseline body mass index (calculated as the weight in kilograms divided by the square of height in meters), depressed late adolescent girls were at a greater than 2-fold increased risk for obesity in adulthood compared with their nondepressed female peers (relative risk, 2.32; 95% confidence interval, 1.29-3.83). A dose-response relationship between the number of episodes of depression during adolescence and risk for adult obesity was also observed in female subjects. The association was not observed for late adolescent boys or for early adolescent boys or girls. CONCLUSIONS: Depression in late adolescence is associated with later obesity, but only among girls. Future studies should address reasons for these age and sex differences and the potential for intervention to reduce the risk for adult obesity in depressed older adolescent girls. PMID- 12912779 TI - Social marginalization of overweight children. AB - BACKGROUND: Overweight is the most common health problem that faces children and adolescents. Although the correlation among overweight, low self-esteem, and depression is well known, social isolation among overweight children and adolescents has not been studied. OBJECTIVE: To investigate social networks of overweight and normal-weight adolescents in a large, nationally representative sample. DESIGN: Cross-sectional, nationally representative cohort study. Population A total of 90 118 adolescents aged 13 to 18 years who were enrolled in the National Longitudinal Study of Adolescent Health, of which a 1:5 subsample was selected for detailed in-home assessment, including height and weight measurements (n = 17 557). Overweight was defined according to body mass index (>95th percentile for age and sex). MAIN OUTCOME MEASURES: This analysis focuses on the number of friendship nominations each adolescent received from other adolescents. The number of friendship nominations and other social network measures were calculated using statistical software. RESULTS: Overweight adolescents were more likely to be socially isolated and to be peripheral to social networks than were normal-weight adolescents. Although overweight adolescents listed similar numbers of friends as normal-weight adolescents, overweight adolescents received significantly fewer friendship nominations from others than were received by normal-weight adolescents (mean [SE] number of friendship nominations, 3.39 [0.08] vs 4.79 [0.04]; P<.001). Overweight adolescents were also more likely to receive no friendship nominations than were normal-weight adolescents (odds ratio, 1.71; 95% confidence interval, 1.39-2.20). Decreased television viewing (P<.001), increased levels of sports participation (P<.001), and increased participation in school clubs (P<.001) were associated with significantly more friendship nominations and higher network centrality scores among both overweight and normal-weight adolescents. CONCLUSIONS: Many overweight adolescents are socially marginalized. Such isolation may aggravate the social and emotional consequences of overweight in this age group. PMID- 12912780 TI - Body satisfaction in 8-year-old children after long-term dietary counseling in a prospective randomized atherosclerosis prevention trial. AB - OBJECTIVE: To investigate whether dietary counseling designed for primary prevention of atherosclerosis and given repeatedly since infancy had an effect on prepubertal children's body satisfaction. DESIGN: Randomized controlled trial. PARTICIPANTS: At the age of 7 months, 1062 infants were randomized to an intervention group (n = 540) or a control group (n = 522). At the age of 8 years, body satisfaction of 217 children in the intervention group and 218 in the control group was evaluated.Intervention Since the children were 8 months old, families in the intervention group had regularly received individualized health education and dietary advice aimed at decreasing the children's intake of saturated fat and cholesterol. MAIN OUTCOME MEASURES: A pictorial instrument was used in measuring estimated current and desired body sizes; a difference between the 2 indicated body dissatisfaction. Weight and height were measured. RESULTS: When adjusted for relative weight, there were no differences in the mean values of estimated current size, desired size, or body dissatisfaction between the girls in the intervention and control groups (P =.62, P =.72, and P =.39, respectively), or between the boys in the intervention and control groups (P =.21, P =.64, and P =.53, respectively). The proportions of children who were satisfied with their size, who wished to be thinner, or who wished to look heavier did not differ between the intervention and control groups in either girls (P =.65) or boys (P =.85). CONCLUSION: Long-term, individualized dietary counseling since infancy with the focus on dietary fat did not enhance body dissatisfaction or desire to be thinner in 8-year-old children. PMID- 12912781 TI - Dietary composition and weight change among low-income preschool children. AB - OBJECTIVE: To examine the relation between dietary composition and weight change among children. We tested several hypotheses considering intake of nutrients (total fat and fiber) and predefined food groups (breads and grains, "fat foods," fruits, and vegetables) used in the North Dakota Special Supplemental Nutrition Program for Women, Infants, and Children (WIC Program). DESIGN: Prospective study. Subjects We collected dietary, anthropometric, and sociodemographic data from 1379 children aged 2 to 5 years participating in the North Dakota WIC Program on 2 visits ranging from 6 to 12 months apart. Main Outcome Measure Annual change in weight. RESULTS: In multiple regression analyses, no significant relations were found between total intake of fat, fiber, fruits, or vegetables and weight change. There was a 0.16-kg lower weight change per year (95% confidence interval [CI], -0.20 to -0.12 kg; P<.01) with each additional daily serving of breads and grains, and a 0.05-kg greater weight change per year (95% CI, 0.1-0.09 kg; P<.05) for each additional serving of fat foods in a model adjusting for sex, age, baseline weight, change in height, and sociodemographic variables. CONCLUSIONS: Intake of North Dakota WIC Program-defined fat foods, but not dietary fat per se, significantly predicted weight gain, whereas intake of North Dakota WIC Program-defined breads and grains, but not fiber per se, significantly predicted weight loss in preschool children. PMID- 12912782 TI - Promoting healthy weight among elementary school children via a health report card approach. AB - BACKGROUND: As overweight continues to rise among children, schools seek effective and sensitive ways to engage parents in promoting healthy weight. OBJECTIVE: To evaluate a school-based health report card on the family awareness of and concern about the child weight status, plans for weight control, and preventive behaviors. DESIGN: Quasi-experimental field trial with a personalized weight and fitness health report card intervention (PI), a general-information intervention (GI), and a control group (CG). Outcomes were assessed using a postintervention telephone survey, including process and outcome measures. PARTICIPANTS: The intervention included 1396 ethnically diverse students at 4 elementary schools in an urban area. Telephone surveys were completed by 399 families from an evaluation sample of 793. Intervention Families were randomly assigned to the PI, GI, or CG and mailed intervention materials. The CG was mailed GI materials after the survey. MAIN OUTCOME MEASURES: Parent awareness of child weight status, concerns, weight-control plans, and preventive behaviors. Group effects were significantly different by the child's weight status, so results were stratified. RESULTS: Among overweight students, intervention parents were more likely to know their child's weight status (PI, 44%; GI, 41%; CG, 23%) (P =.02). The PI parents planned medical help (PI, 25%; GI, 7%; CG, 9%) (P =.004), dieting activities (PI, 19%; GI and CG, <5 cases) (P =.02) and physical activities (PI, 42%; GI, 27%; CG, 13%) (P<.001) for their overweight children. No group effect on concern or preventive behaviors was detected. Most parents of overweight children who read materials requested annual weight and health information on their child (PI, 91%; GI, 67%). CONCLUSIONS: Among overweight children, the PI was associated with increased parental awareness of their child's weight status. Although parents wanted PI for their children, more research is needed to test this approach on children's self-esteem and plans for weight control. PMID- 12912783 TI - A reduced-glycemic load diet in the treatment of adolescent obesity. AB - BACKGROUND: The incidence of type 2 diabetes increases markedly for obese children after puberty. However, the effect of dietary composition on body weight and diabetes risk factors has not been studied in adolescents. OBJECTIVE: To compare the effects of an ad libitum, reduced-glycemic load (GL) diet with those of an energy-restricted, reduced-fat diet in obese adolescents. DESIGN: Randomized control trial consisting of a 6-month intervention and a 6-month follow-up. MAIN OUTCOME MEASURES: Body composition (body mass index [BMI; calculated as weight in kilograms divided by the square of height in meters] and fat mass) and insulin resistance (homeostasis model assessment) were measured at 0, 6, and 12 months. Seven-day food diaries were used as a process measure. SUBJECTS: Sixteen obese adolescents aged 13 to 21 years. Intervention Experimental (reduced-GL) treatment emphasized selection of foods characterized by a low to moderate glycemic index, with 45% to 50% of energy from carbohydrates and 30% to 35% from fat. In contrast, conventional (reduced-fat) treatment emphasized selection of low-fat products, with 55% to 60% of energy from carbohydrates and 25% to 30% from fat. RESULTS: Fourteen subjects completed the study (7 per group). The GL decreased significantly in the experimental group, and dietary fat decreased significantly in the conventional group (P<.05 for both). At 12 months, mean +/- SEM BMI (-1.3 +/- 0.7 vs 0.7 +/- 0.5; P =.02) and fat mass (-3.0 +/- 1.6 vs 1.8 +/- 1.0 kg; P =.01) had decreased more in the experimental compared with the conventional group, differences that were materially unchanged in an intention-to-treat model (n = 16) (BMI, P =.02; fat mass, P =.01). Insulin resistance as measured by means of homeostasis model assessment increased less in the experimental group during the intervention period (-0.4 +/- 0.9 vs 2.6 +/- 1.2; P =.02). In post hoc analyses, GL was a significant predictor of treatment response among both groups (R2 = 0.51; P =.006), whereas dietary fat was not (R2 = 0.14; P =.22). CONCLUSIONS: An ad libitum reduced-GL diet appears to be a promising alternative to a conventional diet in obese adolescents. Large-scale randomized controlled trials are needed to further evaluate the effectiveness of reduced-GL and -glycemic index diets in the treatment of obesity and prevention of type 2 diabetes. PMID- 12912784 TI - Lower risk of overweight in school-aged food insecure girls who participate in food assistance: results from the panel study of income dynamics child development supplement. AB - BACKGROUND: A quarter of US children are at increased risk of obesity-related health problems because of excess weight. The few national studies of child overweight and inadequate household resources available to purchase healthy foods, or food insecurity, have shown inconsistent results. We hypothesized that the inconsistency in these results may be attributed to a differential effect of participation in food assistance programs in food secure and food insecure households. METHODS: Using the 1997 Panel Study of Income Dynamics Child Development Supplement, we compared the risk of a child aged 5 to 12 years being at or above the 85th percentile of body mass index for age in food insecure and food secure, low-income households when controlling for participation in the Food Stamp Program and the national school lunch and breakfast programs as well as other covariates. RESULTS: We found that food insecure girls who participated in all 3 programs had a 68% reduced odds (odds ratio, 0.32; 95% confidence interval, 0.12-0.77) of being at risk of overweight when compared with food insecure girls in nonparticipating households and controlling for other factors. Girls in food secure households generally had no greater or less risk of overweight if they participated in any or all programs. Boys in both food insecure and food secure households had no greater or less risk of overweight by if they participated in any or all of the programs. CONCLUSIONS: These results support our hypothesis that food assistance programs play a protective role for low-income children's health, particularly in girls in food insecure households. PMID- 12912785 TI - Reported consumption of low-nutrient-density foods by American children and adolescents: nutritional and health correlates, NHANES III, 1988 to 1994. AB - OBJECTIVE: To examine the contribution of foods of modest nutritional value to the diets of American children and adolescents. METHODS: The data were from the third National Health and Nutrition Examination Survey, 1988 to 1994, and included 4852 children and adolescents, aged 8 to 18 years. Foods reported in the 24-hour dietary recall were grouped into the following low-nutrient-density (LND) food categories: visible fat; table sweeteners, candy, and sweetened beverages; baked and dairy desserts; salty snacks; and miscellaneous. The independent association of the number of LND foods mentioned in the recall with intake of food groups, macronutrients, micronutrients, and body mass index was examined by means of regression procedures to adjust for multiple covariates. RESULTS: The LND foods contributed more than 30% of daily energy, with sweeteners and desserts jointly accounting for nearly 25%. Intakes of total energy and percentage of energy from carbohydrate and fat related positively, but percentage of energy from protein and dietary fiber (in grams) related inversely to the reported number of LND foods (P<.05). The reported number of LND foods was a negative predictor (P<.001) of the amount of nutrient-dense foods reported. The mean amount of reported intake of several micronutrients-vitamins A, B6, and folate, and the minerals calcium, magnesium, iron, and zinc-declined (P<.05) with increasing tertiles of reported number of LND foods. The LND food reporting was not a significant predictor of body mass index. CONCLUSION: High LND food reporting was related to higher energy intake but lower amounts of the 5 major food groups and most micronutrients. PMID- 12912786 TI - Is physical activity differentially associated with different types of sedentary pursuits? AB - OBJECTIVE: To determine whether there is a relationship between the time adolescents spend in physical activity and time they spend in different sedentary pursuits: watching television, playing video games, working on computers, doing homework, and reading, taking into account the effect of part-time work on students' residual time. DESIGN: Cross-sectional cohort design. PARTICIPANTS AND SETTING: Seven hundred forty-three high school students from 2 inner-city public schools and 1 private school. METHODS: Students completed a self-administered questionnaire that addressed time spent in physical activity, time spent in sedentary pursuits, musculoskeletal pain, and psychosocial issues and were also measured for height and weight. Main Outcome Measure Level of physical activity (low, moderate, high). RESULTS: There were more girls than boys in the low and moderate physical activity groups and more boys than girls in the high activity group. Ordinal logistic regression showed that increased time spent in "productive sedentary behavior" (reading or doing homework and working on computers) was associated with increased physical activity (odds ratio, 1.7; 95% confidence interval, 1.2-2.4), as was time spent working (odds ratio, 1.3; 95% confidence interval, 1.2-1.4). Time spent watching television and playing video games was not associated with decreased physical activity. CONCLUSIONS: Physical activity was not inversely associated with watching television or playing video games, but was positively associated with productive sedentary behavior and part time work. Some students appear capable of managing their time better than others. Future studies should explore the ability of students to manage their time and also determine what characteristics are conducive to better time management. PMID- 12912787 TI - Factors associated with changes in physical activity: a cohort study of inactive adolescent girls. AB - OBJECTIVE: To identify factors associated with changes in physical activity in adolescent girls at risk for sedentary lifestyles and obesity. DESIGN: A cohort study was performed with 201 high school girls recruited to participate in an evaluation study of a school-based obesity prevention physical education program. Three assessments were performed during an 8-month period. MAIN OUTCOME MEASURES: Associations between physical activity and a range of personal factors (self acceptance, self-worth, athletic competence, body image, depressive mood, perceived benefits, enjoyment of physical activity, self-efficacy, and body mass index), behavioral factors (watching television and time constraints), and socioenvironmental factors (social support and costs/resources) were assessed. RESULTS: The 2 strongest and most consistent factors associated with change in physical activity were time constraints and support for physical activity from peers, parents, and teachers. Measures assessing self-perceptions, global (ie, self-worth) and specific to physical activity (ie, self-efficacy to be physically active), were also associated with change in physical activity. For example, a decrease of 2.0 U for an adolescent's perceived time constraints (possible range, 3.0-12.0 U) would be expected to lead to an increase of 53 minutes of moderate to vigorous physical activity per week (95% confidence interval, 33-72 minutes). An increase of 2.0 U in perceived support for physical activity (possible range, 3.0 12.0 U) would be expected to lead to an increase of 35 minutes of moderate to vigorous physical activity per week (95% confidence interval, 13-56 minutes). An increase of 3.0 U on the self-worth scale (possible range, 5.0-20.0 U) might be expected to lead to an increase of 19 minutes of moderate to vigorous physical activity per week (95% confidence interval, 0-40 minutes). CONCLUSION: The effectiveness of interventions aimed at increasing physical activity among adolescent girls might be enhanced by engaging support from friends, family, and caring adults; addressing real and perceived time constraints; and helping adolescent girls feel more confident about themselves and their ability to engage in physical activity. PMID- 12912788 TI - Physical activity in middle school-aged children participating in a school-based recreation program. AB - BACKGROUND: The increasing incidence of obesity in children may be attributed in part to increased sedentary behavior, such as watching television, which leads to less energy expended in physical activity. We have theorized that by middle school, many children lack the physical skills or self-confidence to participate in competitive physical activities. Thus, we hypothesized that if we provided a summer and after-school program featuring noncompetitive, outdoor activities such as gardening and adventure education, we would observe increased physical activity relative to habitual physical activities at home. METHODS: To test this hypothesis, 2 experiments were conducted. In the first, 4 children aged 12 years were evaluated while they participated in a summer recreation program for 2 hours and again while they watched a videotape. They wore a uniaxial accelerometer to assess physical movement, and we used a bicarbonate labeled with 13C tracer technique to assess energy expenditure. In a second experiment, we evaluated 8 children (aged 10-12 years) twice using uniaxial accelerometry only, once while they attended the after-school program for 2 hours and then during a similar period at home. RESULTS: The first study showed that the estimated energy expenditure (kilocalories. kilograms-1. hours-1) was 60% increased during the program (mean +/- SD) (2.6 +/- 0.5) compared with watching a videotape (1.6 +/- 0.3) (P =.02). Physical movement (accelerations per minute) also was significantly increased (3959 +/- 896 vs 513 +/- 182) (P =.004). In the second experiment, movement was 95% increased during the program (4578 +/- 1004) compared with the behavior at home (2345 +/- 746) (P =.005). CONCLUSION: These results show that an organized, noncompetitive, leisure-time program can increase physical activity in children. PMID- 12912789 TI - Physical activity and body mass index among US adolescents: youth risk behavior survey, 1999. AB - OBJECTIVE: To investigate associations of underweight and overweight with physical activity among high school students in the United States. METHODS: A nationally representative sample of 15 349 US high school students participated in the 1999 Youth Risk Behavior Survey; 13 295 were included in these analyses. Five measures of physical activity were examined as dichotomous variables: (1) vigorous-intensity physical activity (>/=3 vs <3 sessions lasting at least 20 minutes each per week); (2) moderate-intensity physical activity (>/=5 vs <5 sessions lasting at least 30 minutes each per week); (3) strength training (>/=3 vs <3 sessions per week); (4) enrollment in physical education (yes or no); and (5) sports participation (yes or no). Using body mass indexes, students were categorized by percentiles as underweight (5th to 15th to <85th percentiles), at risk for overweight (>/=85th to <95th percentiles), or overweight (>/=95th percentile). Potential associations between physical activity and body mass index were examined using logistic regression. RESULTS: On several measures, adolescent boys who were underweight or overweight were less likely to be physically active than boys of normal weight (eg, odds ratio [OR], 0.23; 95% confidence interval [CI], 0.12-0.45; and OR, 0.75; 95% CI, 0.61-0.93; for boys who were underweight and overweight, respectively, for strength training). Adolescent girls who were overweight or at risk for overweight were less likely (OR, 0.62; 95% CI, 0.50-0.78; and OR, 0.63; 95% CI, 0.46-0.85; respectively) to be involved with sports than girls of normal weight; and girls who were underweight were less likely (OR, 0.44; 95% CI, 0.22-0.91) to be enrolled in physical education. CONCLUSIONS: Weight status among high school students is correlated with selected physical activity behavior, especially among adolescent boys. Interventions to increase physical activity for high school students should target adolescents of all shapes and sizes, and may best be achieved by school policies requiring physical education or after-school sports. PMID- 12912790 TI - Prevalence of a metabolic syndrome phenotype in adolescents: findings from the third National Health and Nutrition Examination Survey, 1988-1994. AB - BACKGROUND: In adults the metabolic syndrome imposes a substantial risk for type 2 diabetes mellitus and premature coronary heart disease. Even so, no national estimate is currently available of the prevalence of this syndrome in adolescents. OBJECTIVE: To estimate the prevalence and distribution of a metabolic syndrome among adolescents in the United States. DESIGN AND SETTING: Analyses of cross-sectional data obtained from the Third National Health and Nutrition Examination Survey (1988-1994), which was administered to a representative sample of the noninstitutionalized civilian population of the United States. PARTICIPANTS: Male and female respondents aged 12 to 19 years (n = 2430). MAIN OUTCOME MEASURES: The prevalence and distribution of a metabolic syndrome among US adolescents, using the National Cholesterol Education Program (Adult Treatment Panel III) definition modified for age. RESULTS: The overall prevalence of the metabolic syndrome among adolescents aged 12 to 19 years was 4.2%; 6.1% of males and 2.1% of females were affected (P=.01). The syndrome was present in 28.7% of overweight adolescents (body mass index [BMI], >/=95th percentile) compared with 6.8% of at-risk adolescents (BMI, 85th to <95th percentile) and 0.1% of those with a BMI below the 85th percentile (P<.001). Based on population-weighted estimates, approximately 910 000 US adolescents have the metabolic syndrome. CONCLUSIONS: Perhaps 4% of adolescents and nearly 30% of overweight adolescents in the United States meet these criteria for a metabolic syndrome, a constellation of metabolic derangements associated with obesity. These findings may have significant implications for both public health and clinical interventions directed at this high-risk group of mostly overweight young people. PMID- 12912791 TI - Is nuclear factor kappaB an attractive therapeutic target for treating cardiac hypertrophy? PMID- 12912792 TI - Plaque rupture, lysophosphatidic acid, and thrombosis. PMID- 12912793 TI - Leptin: a novel link between obesity, diabetes, cardiovascular risk, and ventricular hypertrophy. PMID- 12912794 TI - Clinical use of cardiovascular magnetic resonance. PMID- 12912795 TI - Aortic dissection: new frontiers in diagnosis and management: Part II: therapeutic management and follow-up. PMID- 12912796 TI - Images in cardiovascular medicine. Concomitant T-wave alternans and pulsus alternans in a child with long-QT syndrome. PMID- 12912797 TI - Angina pectoris without chest pain. PMID- 12912798 TI - Caffeine and coffee tolerance. PMID- 12912800 TI - Aortic valve replacement in patients with severe aortic regurgitation and reduced left ventricular ejection fraction. PMID- 12912799 TI - Anemia and congestive heart failure. PMID- 12912801 TI - Cardioprotective effect of diazoxide is mediated by activation of sarcolemmal but not mitochondrial ATP-sensitive potassium channels in mice. PMID- 12912802 TI - Repeating intracoronary radiation an option for in-stent restenosis, even after initial failure. PMID- 12912804 TI - Soluble CD40L: risk prediction after acute coronary syndromes. AB - BACKGROUND: Elevated plasma concentrations of soluble CD40 ligand (sCD40L) indicate increased risk for future cardiovascular events in apparently healthy women. This study tested the hypothesis that plasma sCD40L, alone or in combination with troponin (cTnI) or C-reactive protein (CRP), may identify patients with acute coronary syndromes at heightened risk for recurrent cardiac events. METHODS AND RESULTS: In a nested case-control study (cases, n=195; controls, n=195) within the OPUS-TIMI16 trial, patients with the prespecified study end points death, myocardial infarction (MI), or congestive heart failure (CHF) within 10 months had significantly higher median (25th, 75th percentiles) sCD40L plasma levels than did controls (0.78 [0.34, 1.73] ng/mL versus 0.52 [0.16, 1.42] ng/mL, P<0.002). After adjustment for other risk predictors and levels of cTnI and CRP, sCD40L levels above median were associated with higher risk for death, MI, and the composite death/MI or death/MI/CHF (adjusted hazard ratios, 1.9 [P<0.05], 1.9 [P<0.001], 1.9 [P<0.001], and 1.8 [P<0.01], respectively). Interestingly, patients with elevated plasma levels of sCD40L and cTnI showed a markedly increased risk of death, MI, or death/MI/CHF compared with patients with the lowest levels of both markers (adjusted hazard ratios, 12.1, 7.2, and 4.3, respectively; all P<0.01). CONCLUSIONS: Elevated plasma levels of sCD40L identify patients with acute coronary syndromes at heightened risk of death and recurrent MI independent of other predictive variables, including cTnI and CRP. Notably, combined assessment of sCD40L with cTnI complements prognostic information for death and MI. PMID- 12912803 TI - Induction of a regulatory T cell type 1 response reduces the development of atherosclerosis in apolipoprotein E-knockout mice. AB - BACKGROUND: T helper type 1 (Th1) response plays a permissive role in atherosclerosis. We hypothesized that adoptive transfer of a novel subtype of T lymphocytes called regulatory T cells type 1 (Tr1) would inhibit Th1 responses by inducing a bystander immune suppression and therefore limit the development of atherosclerosis. METHODS AND RESULTS: Clones of ovalbumin (OVA)-specific Tr1 cells expanded in vitro were administered intraperitoneally (106 cells per mouse) with their cognate antigen (50 microg of OVA subcutaneously in complete Freund's adjuvant [CFA]) to female apolipoprotein E-knockout mice. A group of mice received only (OVA/CFA) immunization without Tr1 cells. Two other control groups received no immunization and were injected with either Tr1 cells or saline. After 9 weeks of treatment, mice injected with (OVA/CFA)+OVA-specific Tr1 cells showed a significant decrease in Th1 responses, as revealed by a decrease in OVA specific IgG2a serum levels (P<0.0001), a decrease in the production of interferon-gamma (P<0.001), and an increase in interleukin-10 production (P<0.001) by cultured spleen and lymph T cells compared with controls. In addition, cytokine production by concanavalin A-stimulated spleen cells showed a clear switch to a regulatory immune response in mice treated with (OVA/CFA)+Tr1. This was associated with a significant reduction in atherosclerotic lesion size in both the thoracic aorta and aortic sinus of mice treated with (OVA/CFA)+Tr1 compared with controls (P=0.002 to P<0.0001). Plaques of mice injected with (OVA/CFA)+Tr1 showed significantly lower accumulation of macrophages and T cells than plaques of control mice. CONCLUSIONS: Tr1-type regulatory immune response reduces the development of experimental atherosclerosis. PMID- 12912805 TI - Role of a decreased expression of the local renin-angiotensin system in the etiology of cerebral aneurysms. AB - BACKGROUND: Local renin-angiotensin systems (RAS) have been implicated as playing an important role in vascular remodeling. The relationship of this system to the etiology of cerebral aneurysm was investigated. METHODS AND RESULTS: The aneurysmal wall from patients with a ruptured or unruptured cerebral aneurysm and the cortical cerebral artery in control patients with head trauma or a glioma were taken during surgery for study. Local RAS were evaluated by reverse transcription-polymerase chain reaction (RT-PCR) and/or immunohistochemistry. RT PCR analysis revealed a significantly decreased expression of angiotensin converting enzyme (ACE), angiotensin type 1 (AT1) receptor, basic fibroblast growth factor, platelet-derived growth factor-AA, and tissue inhibitor of matrix metalloproteinases-1 mRNA in the aneurysmal wall as compared with the control cortical arterial wall. Immunohistochemistry also revealed a decreased expression of ACE, AT1 receptor, and angiotensin II in the aneurysmal wall. CONCLUSIONS: Expression of local RAS was decreased in the aneurysmal wall, which may induce aneurysm formation caused by a lack of vascular remodeling that prevents the arterial wall from thickening under increased hemodynamic stress. This is the first report that suggests that a decreased expression of local RAS plays a part in the pathogenesis of any disease. PMID- 12912806 TI - Hemodynamic profile after the Norwood procedure with right ventricle to pulmonary artery conduit. AB - BACKGROUND: The balance of systemic, pulmonary, and coronary blood flow after the Norwood operation for hypoplastic left heart syndrome (HLHS) is critical to early survival. We hypothesized that a right ventricle to pulmonary artery conduit (instead of a systemic to pulmonary artery shunt) would result in hemodynamic changes consistent with a more stable balance of systemic, pulmonary, and coronary perfusion. METHODS AND RESULTS: Hemodynamic data were obtained during cardiac catheterization before the hemi-Fontan procedure from 24 patients with HLHS; the first 10 had a Norwood operation with a systemic to pulmonary artery shunt, and the latter 14 had the Norwood operation with a right ventricle to pulmonary artery conduit. Significant differences were present, with the right ventricle to pulmonary artery conduit group having a higher aortic diastolic pressure (55 versus 42 mm Hg), a narrowed systemic pulse pressure (43 versus 64 mm Hg), a lower Qp:Qs (0.92 versus 1.42), a higher coronary perfusion pressure (46 versus 32 mm Hg), and a higher ratio of pulmonary artery diameter to descending aorta diameter (1.51 versus 1.37). CONCLUSIONS: We conclude that, in HLHS after the Norwood operation, the right ventricle to pulmonary artery conduit modification produces hemodynamic changes consistent with improved coronary perfusion and a more favorable distribution of systemic, pulmonary, and coronary blood flow. PMID- 12912807 TI - Antioxidant vitamins C and E improve endothelial function in children with hyperlipidemia: Endothelial Assessment of Risk from Lipids in Youth (EARLY) Trial. AB - BACKGROUND: Hyperlipidemia is associated with endothelial dysfunction, an early event in atherosclerosis and predictor of risk for future coronary artery disease. Epidemiological studies suggest that increased dietary intake of antioxidants reduces the risk of coronary artery disease. The purpose of this study was to determine whether antioxidant vitamin therapy improves endothelial function and affects surrogate biomarkers for oxidative stress and inflammation in hyperlipidemic children. METHODS AND RESULTS: In a randomized, double-blind, placebo-controlled trial, the effects of antioxidant vitamins C (500 mg/d) and E (400 IU/d) for 6 weeks and the National Cholesterol Education Program Step II (NCEP-II) diet for 6 months on endothelium-dependent flow-mediated dilation (FMD) of the brachial artery were examined in 15 children with familial hypercholesterolemia (FH) or the phenotype of familial combined hyperlipidemia (FCH). Antioxidant vitamin therapy improved FMD of the brachial artery compared with baseline (P<0.001) without an effect on biomarkers for oxidative stress (autoantibodies to epitopes of oxidized LDL, F2-isoprostanes, 8-hydroxy-2' deoxyguanosine), inflammation (C-reactive protein), or levels of asymmetric dimethylarginine, an endogenous inhibitor of nitric oxide. CONCLUSIONS: Antioxidant therapy with vitamins C and E restores endothelial function in hyperlipidemic children. Early detection and treatment of endothelial dysfunction in high-risk children may retard the progression of atherosclerosis. PMID- 12912808 TI - The receptor RAGE as a progression factor amplifying arachidonate-dependent inflammatory and proteolytic response in human atherosclerotic plaques: role of glycemic control. AB - BACKGROUND: RAGE (receptor for advanced glycation end products [AGEs]) plays a role in diabetic atherosclerosis. Recently, we have demonstrated enhanced expression of cyclooxygenase-2 and PGE synthase-1 (COX-2/mPGES-1) in human symptomatic plaques, and provided evidence that it is associated with metalloproteinase (MMP)-induced plaque rupture. However, the specific transmembrane signaling pathway(s) influencing plaque COX-2/mPGES-1 expression is unknown. The aim of this study was to characterize RAGE expression in human plaques and to correlate it with the inflammatory infiltration, COX-2/mPGES-1 and MMP expression, and with clinical evidence of diabetes. METHODS AND RESULTS: Plaques obtained from 60 patients undergoing carotid endarterectomy were divided into diabetic and nondiabetic according to clinical evidence of type 2 diabetes. Plaques were subjected to analysis of RAGE, NF-kappaB, COX-2/mPGES-1, MMP-2 and MMP-9, lipid and oxidized LDL (oxLDL) content, and collagen content by immunohistochemistry and Western blot, whereas zymography was used to detect MMP activity. Immunohistochemistry was used to identify CD68+ macrophages, CD3+ T lymphocytes, smooth muscle cells (SMCs), and HLA-DR+ inflammatory cells. Diabetic plaques had more (P<0.0001) macrophages, T-lymphocytes, and HLA-DR+ cells, more (P<0.0001) immunoreactivity for RAGE, activated NF-kappaB, COX-2/mPGES-1, and MMPs, increased (P<0.0001) gelatinolytic activity, reduced (P<0.0001) collagen content, and increased (P<0.0001) lipid and oxLDL content. Interestingly, RAGE, COX-2/mPGES-1, and MMP expression was linearly correlated with plasma level of HbA1c. CONCLUSIONS: In conclusion, this study demonstrates in humans that RAGE overexpression is associated with enhanced inflammatory reaction and COX-2/mPGES 1 expression in diabetic plaque macrophages, and this effect may contribute to plaque destabilization by inducing culprit metalloproteinase expression. PMID- 12912809 TI - Seasonal variation of fibrinogen in dipper and nondipper hypertensive patients. AB - BACKGROUND: A seasonal variation with higher values in winter has been previously reported in plasma fibrinogen, a recognized marker of the potential risk of myocardial infarction and stroke. The lack of nocturnal decline in blood pressure has also been associated with an increase in cardiovascular events. Accordingly, we have compared the yearly variation of plasma fibrinogen in dipper and nondipper hypertensive patients. METHODS AND RESULTS: We studied 1006 stage 1 to 2 hypertensive patients (482 men and 524 women, 53.0+/-13.4 years of age). Blood pressure was measured every 20 minutes during the day and every 30 minutes at night for 48 consecutive hours. Physical activity was simultaneously evaluated at 1-minute intervals with a wrist actigraph. A blood sample was collected on the same day before starting blood pressure monitoring. The circannual variation of fibrinogen was established for all patients as well as for subgroups of dippers and nondippers (n=513; nocturnal blood pressure decline <10%) by multiple component analysis. For the whole group of patients, fibrinogen was characterized by a highly significant seasonal variation (P<0.001) with a mean value of 318 mg/dL, double circannual amplitude (extent of predictable change along the year) of 40 mg/dL, and time of peak value in February. Throughout the year, the nondippers showed higher plasma fibrinogen levels than did the dippers (P<0.001). CONCLUSIONS: The elevated plasma fibrinogen levels in nondipper patients appear to be directly related to their increased risk in vascular events, which are more prominent during the late winter months. PMID- 12912810 TI - Dual functionality of cyclooxygenase-2 as a regulator of tumor necrosis factor mediated G1 shortening and nitric oxide-mediated inhibition of vascular smooth muscle cell proliferation. AB - BACKGROUND: Cyclooxygenase (COX)-2 contributes to vascular smooth muscle cell (VSMC) proliferation induced by tumor necrosis factor (TNF) and angiotensin II. The present study demonstrates, however, that depending on prevailing conditions, COX-2-derived prostanoids may also inhibit VSMC proliferation. METHODS AND RESULTS: TNF-alpha stimulated proliferation of VSMCs by shortening the G1 phase of the cell cycle. This effect was abolished by NS-398, a selective COX-2 inhibitor. Addition of TNF did not affect the protein-to-DNA ratio, measured by flow cytometry, suggesting that TNF does not induce VSMC hypertrophy. Inhibition of nitric oxide synthase (NOS) activity attenuated TNF-mediated increases in prostaglandin (PG) I2 synthesis, whereas thromboxane (TX) A2 production and COX-2 protein expression were unaffected. Moreover, inhibition of NOS activity increased TNF-mediated proliferation by approximately 23%. Thus, NO preferentially stimulates PGI2 production, suggesting that production of NO by VSMCs challenged with TNF limits the ability of the cytokine to increase proliferation. NO donors increased COX-2 protein expression and PGI2 synthesis, had no effect on TXA2 production, and decreased cell numbers by 50%, indicating that expression of COX-2 per se might not be sufficient to support proliferation. The effects of NO donors were prevented when COX-2 activity was inhibited with NS 398. CONCLUSIONS: The COX-2-dependent proliferative response of VSMCs to TNF was modulated in an NO-dependent manner, and PGI2 derived from COX-2 might contribute to the antiproliferative effect of NO donors. PMID- 12912811 TI - Targeted overexpression of transmembrane tumor necrosis factor provokes a concentric cardiac hypertrophic phenotype. AB - BACKGROUND: Tumor necrosis factor (TNF) is initially synthesized as a 26-kDa transmembrane protein that is enzymatically cleaved by TNF-alpha converting enzyme (TACE) to generate a 17-kDa form of "secreted" TNF. Whereas the effects of secreted TNF in the heart have been characterized extensively, the effects of transmembrane TNF in the heart are unknown. METHODS AND RESULTS: We generated lines of transgenic mice with cardiac-restricted overexpression of a noncleavable, transmembrane form of TNF. We next treated a previously generated transgenic line of mice with cardiac-restricted expression of cleavable TNF (referred to as MHCsTNF mice) with a TACE inhibitor (DPC-IDR1) to determine whether TACE inhibition would prevent the transition from concentric hypertrophy to left ventricular (LV) dilation that occurs in this line of transgenic mice. Two of the founder lines did not have a demonstrable phenotype (M-41 and M-45), whereas a third line developed a concentric hypertrophic cardiac phenotype (M 48). Characterization of the M-48 line at 6 weeks of age showed that this line developed concentric hypertrophy, with an increase in myocyte cross-sectional area and reexpression of the fetal gene program. Four weeks of TACE inhibition abrogated the LV dilation in the MHCsTNF mice and resulted in an increase in LV wall thickness and increased myocyte cross-sectional area, thus mimicking the effects observed in the mice with noncleavable, transmembrane TNF. CONCLUSIONS: These studies show that transmembrane TNF is biologically active and provokes a concentric hypertrophic cardiac phenotype, thus suggesting that posttranslational processing (ie, secretion) of TNF is responsible for the dilated cardiomyopathic phenotype in mice with targeted, cardiac-restricted overexpression of TNF. PMID- 12912812 TI - Impact of valve prosthesis-patient mismatch on short-term mortality after aortic valve replacement. AB - BACKGROUND: The prosthesis used for aortic valve replacement (AVR) can be too small in relation to body size, thus causing valve prosthesis-patient mismatch (PPM) and abnormally high transvalvular pressure gradients. This study examined if there is a relation between PPM and short-term mortality after operation. METHODS AND RESULTS: The indexed valve effective orifice area (EOA) was estimated for each type and size of prosthesis being implanted in 1266 consecutive patients and used to define PPM as not clinically significant if >0.85 cm2/m2, as moderate if >0.65 cm2/m2 and 120 minutes (P=0.001), and PPM (P=0.003). Relative risk of mortality was increased 2.1-fold (95% confidence interval, 1.2 to 3.7) in patients with moderate PPM and 11.4-fold (4.4 to 29.5) in those with severe PPM. Moreover, risk of mortality for every category of PPM was higher in patients with a left ventricular ejection fraction <40% as compared with >or=40% (nonsignificant PPM, 2.7 versus 1.0; moderate PPM, 7.1 versus 1.8; severe PPM, 77.1 versus 11.3). CONCLUSIONS: PPM is a strong and independent predictor of short-term mortality among patients undergoing AVR, and its impact is related both to its degree of severity and the status of left ventricular function. In contrast to other risk factors, moderate-severe PPM can be largely avoided with the use of a prospective strategy at the time of operation. PMID- 12912813 TI - Natural history of asymptomatic left ventricular systolic dysfunction in the community. AB - BACKGROUND: Information is limited regarding the rates of progression to congestive heart failure (CHF) and death in individuals with asymptomatic left ventricular systolic dysfunction (ALVD). We sought to characterize the natural history of ALVD, by studying unselected individuals with this condition in the community. METHODS AND RESULTS: We studied 4257 participants (1860 men) from the Framingham Study who underwent routine echocardiography. The prevalence of ALVD (visually estimated ejection fraction [EF]50%, n=4128) and ALVD (n=129) were 0.7 and 5.8 per 100 person-years, respectively. After adjustment for cardiovascular disease risk factors, ALVD was associated with a hazards ratio (HR) for CHF of 4.7 (95% CI 2.7 to 8.1), compared with individuals without ALVD. An elevated risk of CHF after ALVD was observed even in individuals without prior myocardial infarction or valvular disease, with an adjusted HR of 6.5 (CI 3.1 to 13.5). Mild ALVD (EF 40% to 50%, n=78) and moderate-to-severe ALVD (EF <40%, n=51) were associated with adjusted HRs for CHF of 3.3 (CI 1.7 to 6.6) and 7.8 (CI 3.9 to 15.6), respectively. ALVD was also associated with an increased mortality risk (adjusted HR 1.6, CI 1.1 to 2.4). The median survival of ALVD subjects was 7.1 years. CONCLUSIONS: Individuals with ALVD in the community are at high risk of CHF and death, even when only mild impairment of EF is present. Additional studies are needed to define optimal therapy for mild ALVD. PMID- 12912814 TI - Platelet/endothelial biomarkers in depressed patients treated with the selective serotonin reuptake inhibitor sertraline after acute coronary events: the Sertraline AntiDepressant Heart Attack Randomized Trial (SADHART) Platelet Substudy. AB - BACKGROUND: Depression after acute coronary syndromes (ACSs) has been identified as an independent risk factor for subsequent cardiac death. Enhanced platelet activation has been hypothesized to represent 1 of the mechanisms underlying this association. Selective serotonin reuptake inhibitors (SSRIs) are known to inhibit platelet activity. Whether treatment of depressed post-ACS patients with SSRIs alters platelet function was not known. Accordingly, we serially assessed the release of established platelet/endothelial biomarkers in patients treated with sertraline vs placebo in the Sertraline AntiDepressant Heart Attack Randomized Trial (SADHART). METHODS AND RESULTS: Plasma samples (baseline, week 6, and week 16) were collected from patients randomized to sertraline (n=28) or placebo (n=36). Anticoagulants, aspirin, and ADP-receptor inhibitors were permitted in this study. Platelet factor 4, beta-thromboglobulin (betaTG), platelet/endothelial cell adhesion molecule-1, P-selectin, thromboxane B2, 6 ketoprostaglandin F1a, vascular cell adhesion molecule-1, and E-selectin were measured by ELISA. Treatment with sertraline was associated with substantially less release of platelet/endothelial biomarkers than was treatment with placebo. These differences attained statistical significance for betaTG (P=0.03) at weeks 6 and 16 and for P-selectin (P=0.04) at week 16. Repeated-measures ANOVA revealed a significant advantage for sertraline vs placebo for diminishing E-selectin and betaTG concentrations across the entire treatment period. CONCLUSIONS: Treatment with sertraline in depressed post-ACS patients is associated with reductions in platelet/endothelial activation despite coadministration of widespread antiplatelet regimens including aspirin and clopidogrel. The antiplatelet and endothelium-protective properties of SSRIs might represent an attractive additional advantage in patients with depression and comorbid coronary artery and/or cerebrovascular disease. PMID- 12912815 TI - Adenosine diphosphate-induced platelet aggregation is associated with P2Y12 gene sequence variations in healthy subjects. AB - BACKGROUND: The adenosine diphosphate (ADP) receptor P2Y12 plays a pivotal role in platelet aggregation, as demonstrated by the benefit conferred by its blockade in patients with cardiovascular disease. Some studies have shown interindividual differences in ADP-induced platelet aggregation responses ex vivo, but the mechanisms underlying this variability are unknown. METHODS AND RESULTS: We examined ADP-induced platelet aggregation responses in 98 healthy volunteers, and we identified 2 phenotypic groups of subjects with high and low responsiveness to 2 micromol/L ADP. This prompted us to screen the recently identified Gi-coupled ADP receptor gene P2Y12 for sequence variations. Among the 5 frequent polymorphisms thus identified, 4 were in total linkage disequilibrium, determining haplotypes H1 and H2, with respective allelic frequencies of 0.86 and 0.14. The number of H2 alleles was associated with the maximal aggregation response to ADP in the overall study population (P=0.007). Downregulation of the platelet cAMP concentration by ADP was more marked in 10 selected H2 carriers than in 10 noncarriers. CONCLUSIONS: In healthy subjects, ADP-induced platelet aggregation is associated with a haplotype of the P2Y12 receptor gene. Given the crucial role of the P2Y12 receptor in platelet functions, carriers of the H2 haplotype may have an increased risk of atherothrombosis and/or a lesser clinical response to drugs inhibiting platelet function. PMID- 12912816 TI - Comparative effects of carvedilol and metoprolol on regional vascular responses to adrenergic stimuli in normal subjects and patients with chronic heart failure. AB - BACKGROUND: Adrenergic receptor blockers used in the treatment of heart failure have distinct receptor affinity profiles. We hypothesized that alpha-adrenergic blocking effects of carvedilol would limit vasoconstriction in response to adrenergic stimuli when compared with metoprolol. METHODS AND RESULTS: Forearm vascular resistance responses to isometric handgrip and cold pressor test were determined by plethysmography before and during adrenergic receptor blockade in prospective randomized trials. Acute effects were assessed in a crossover trial in normal subjects (single dose of 25 mg carvedilol, 100 mg metoprolol tartrate, and placebo). Chronic effects (25 mg carvedilol BID versus 200 mg extended release metoprolol succinate daily for 6 months) were assessed in a parallel group trial of chronic heart failure subjects. In normal subjects, carvedilol decreased forearm vascular resistance responses to adrenergic stimuli when compared with metoprolol and placebo (isometric handgrip -3.5 U for carvedilol versus -1.2 U for metoprolol and -2.2 U for placebo, P=0.15; cold pressor test 3.1+/-8.9 U for carvedilol versus 9.0+/-2.7 U for metoprolol and 8.2+/-5.8 U for placebo, P<0.05). In heart failure subjects, vasomotor responses to isometric handgrip and cold pressor test did not differ between treatment groups. CONCLUSIONS: Acute administration of carvedilol attenuates the vasoconstriction response to adrenergic stimuli when compared with placebo and metoprolol in normal subjects, whereas chronic administration of carvedilol does not attenuate the vasoconstrictor response to adrenergic stimuli when compared with metoprolol in heart failure subjects. These data suggest that long-term benefits of carvedilol in heart failure are not mediated by alpha-adrenergic blockade. PMID- 12912817 TI - Relation between hospital intra-aortic balloon counterpulsation volume and mortality in acute myocardial infarction complicated by cardiogenic shock. AB - BACKGROUND: Increasing evidence suggests an inverse relationship between outcome and the total number of invasive cardiac procedures performed at a given hospital. The purpose of the present study was to determine if a similar relationship exists between the number of intra-aortic balloon counterpulsation (IABP) procedures performed at a given hospital per year and the in-hospital mortality rate of patients with acute myocardial infarction complicated by cardiogenic shock. METHODS AND RESULTS: We analyzed data of 12 730 patients at 750 hospitals enrolled in the National Registry of Myocardial Infarction 2 from 1994 to 1998. The hospitals were divided into tertiles (low-, intermediate-, and high-IABP volume hospitals) according to the number of IABPs performed at the given hospital per year. The median number of IABPs performed per hospital per year was 3.4, 12.7, and 37.4 IABPs at low-, intermediate-, and high-volume hospitals, respectively. Of those patients who underwent IABP, there were only minor differences in baseline patient characteristics between the 3 groups. Crude mortality rate decreased with increasing IABP volume: 65.4%, lowest volume tertile; 54.1%, intermediate volume tertile; and 50.6%, highest volume tertile (P for trend <0.001). This mortality difference represented 150 fewer deaths per 1000 patients treated at the high IABP hospitals. In the multivariate analysis, high hospital IABP volume for patients with acute myocardial infarction was associated with lower mortality (OR=0.71, 95% CI=0.56 to 0.90), independent of baseline patient characteristics, hospital factors, treatment, and procedures such as PTCA. CONCLUSIONS: Among the myocardial infarction patients with cardiogenic shock who underwent IABP placement, mortality rate was significantly lower at high-IABP volume hospitals compared with low-IABP volume hospitals. PMID- 12912818 TI - Endothelial dysfunction induced by hyperhomocyst(e)inemia: role of asymmetric dimethylarginine. AB - BACKGROUND: Endothelial function is impaired by hyperhomocyst(e)inemia. We have previously shown that homocyst(e)ine (Hcy) inhibits NO production by cultured endothelial cells by causing the accumulation of asymmetric dimethylarginine (ADMA). The present study was designed to determine if the same mechanism is operative in humans. METHODS AND RESULTS: We studied 9 patients with documented peripheral arterial disease (6 men; 3 women; age, 64+/-3 years), 9 age-matched individuals at risk for atherosclerosis (older adults; 9 men; age, 65+/-1 years), and 5 young control subjects (younger adults; 5 men; age, 31+/-1 years) without evidence of or risk factors for atherosclerosis. Endothelial function was measured by flow-mediated vasodilatation of the brachial artery before and 4 hours after a methionine-loading test (100 mg/kg body weight, administered orally). In addition, blood was drawn at both time points for measurements of Hcy and ADMA concentrations. Plasma Hcy increased after the methionine-loading test in each group (all, P<0.001). Plasma ADMA levels rose in all subjects, from 0.9+/ 0.2 to 1.6+/-0.2 micromol/L in younger adults, from 1.5+/-0.2 to 3.0+/-0.4 micromol/L in older adults, and from 1.8+/-0.1 to 3.9+/-0.3 micromol/L in peripheral arterial disease patients (all, P<0.001). Flow-mediated vasodilatation was reduced from 13+/-2% to 10+/-1% in younger adults, from 6+/-1% to 5+/-1% in older adults, and from 7+/-1% to 3+/-1% in peripheral arterial disease patients (all, P<0.001). Furthermore, we found positive correlations between plasma Hcy and ADMA concentrations (P=0.03, r=0.450), as well as ADMA and flow-mediated vasodilatation (P=0.002, r=0.623). CONCLUSIONS: Our results suggest that experimental hyperhomocyst(e)inemia leads to accumulation of the endogenous NO synthase inhibitor ADMA, accompanied by varying degrees of endothelial dysfunction according to the preexisting state of cardiovascular health. PMID- 12912819 TI - Cisapride-induced transmural dispersion of repolarization and torsade de pointes in the canine left ventricular wedge preparation during epicardial stimulation. AB - BACKGROUND: Cisapride, a gastrointestinal prokinetic agent, was recently withdrawn from the market because of its propensity to induce torsade de pointes (TdP) arrhythmias. The present study examines the electrophysiological actions of cisapride in the isolated arterially perfused canine left ventricular wedge preparation. METHODS AND RESULTS: Transmembrane action potentials from epicardial and M regions and a pseudo-ECG were simultaneously recorded. Cisapride (0.1 to 5 micromol/L) was added to the coronary perfusate. Cisapride prolonged the QT interval and increased transmural dispersion of repolarization (TDR) at relatively low but not at high concentrations. TdP could be induced with programmed electrical stimulation only at a low concentration of drug (0.2 micromol/L), when TDR was maximally prolonged. Moreover, TdP could only be induced during epicardial (but not endocardial) activation of the wedge, which was found to augment TDR. At higher concentrations of cisapride, QT was further prolonged, TDR was diminished, and TdP could no longer be induced. Tpeak-Tend interval and Tpeak-Tend area provided reasonable electrocardiographic indices of TDR. CONCLUSIONS: Our data (1) demonstrate a biphasic concentration/response relationship for the effect of cisapride to induce long-QT syndrome and TdP, (2) show the value of the left ventricular wedge preparation in identifying drugs that pose an arrhythmic risk, (3) support the hypothesis that risk for development of TdP is related to the increase in TDR rather than to prolongation of the QT interval, and (4) indicate that epicardial activation of the left ventricle, as occurs during biventricular pacing, can facilitate the development of TdP under long-QT conditions. PMID- 12912820 TI - Cardiac troponin I is associated with impaired hemodynamics, progressive left ventricular dysfunction, and increased mortality rates in advanced heart failure. AB - BACKGROUND: Cardiac troponin I (cTnI), a sensitive and specific marker of myocardial cell injury, is useful in diagnosing and assessing prognosis in acute coronary syndromes. Small studies report that cTnI is elevated in severe heart failure (HF) and may predict adverse outcomes. METHODS AND RESULTS: The present study evaluated 238 patients with advanced HF referred for cardiac transplantation evaluation who had cTnI assay drawn at the time of initial presentation. Patients with acute myocardial infarction or myocarditis were excluded from analysis. cTnI was detectable (cTnI > or =0.04 ng/mL) in serum of 117 patients (49.1%). Patients with detectable cTnI levels had significantly higher B-type natriuretic peptide (BNP) levels (P<0.001) and more impaired hemodynamic profiles, including higher pulmonary wedge pressures (P=0.002) and lower cardiac indexes (P<0.0001). A significant correlation was found between detectable cTnI and progressive decline in ejection fraction over time. Furthermore, detectable cTnI was associated with increased mortality risk (RR, 2.05; 95% CI, 1.22 to 3.43). After adjustment for other factors associated with adverse prognosis including age, sex, ejection fraction, and coronary artery disease, cTnI remained a significant predictor of death. cTnI used in conjunction with BNP further improved prognostic value. CONCLUSIONS: cTnI is associated with impaired hemodynamics, elevated BNP levels, and progressive left ventricular dysfunction in patients with HF. cTnI may be a novel, useful tool in identifying patients with HF who are at increased risk for progressive ventricular dysfunction and death. PMID- 12912821 TI - Fish consumption is associated with lower heart rates. AB - BACKGROUND: Fish consumption decreases risk of sudden death. The goal of the present study was to assess the relationship between fish consumption and heart rate. METHODS AND RESULTS: A cross-sectional analysis was conducted of 9758 men, age 50 to 59 years, without coronary heart disease (CHD) who were recruited in France and Belfast, Ireland, from 1991 to 1993. Heart rate and CHD risk factors were compared among 4 categories of fish consumption, as follows: (1) less than once per week (n=2662), (2) once per week (n=4576), (3) twice per week (n=1964), and (4) more than twice per week (n=556). Fatty acid profiles of erythrocyte phospholipids were determined in a random subsample of 407 subjects. In erythrocyte phospholipids, eicosapentaenoic acid (P<0.0005), docosahexaenoic acid (P<0.0001), and total n-3 fatty acid (P<0.0008) increased across the categories of fish intake. Triglycerides (P<0.0001), systolic blood pressure (P<0.006), and diastolic blood pressure (P<0.0001) were lower and HDL cholesterol levels (P<0.004) were higher in fish consumers than in nonconsumers. Similarly, heart rate decreased across the categories of fish intake (P<0.0001). After adjustment for age, center, education level, physical activity, smoking habit, alcohol consumption, body mass index, and antiarrhythmic medications, heart rate remained statistically lower among fish consumers than among nonconsumers (P for trend <0.0001). Docosahexaenoic acid content of erythrocyte phospholipids was inversely correlated with heart rate (P<0.03). CONCLUSIONS: Fish consumption is associated with decreased heart rate in men. Because heart rate is positively associated with risk of sudden death, this association may explain, at least in part, the lower risk of sudden death among fish consumers. PMID- 12912823 TI - Ultrasound-targeted microbubble destruction can repeatedly direct highly specific plasmid expression to the heart. AB - BACKGROUND: Noninvasive, tissue-specific delivery of therapeutic agents would be a valuable clinical tool. We have previously shown that ultrasound-targeted microbubble destruction can direct expression of an adenoviral reporter to the heart. The present study shows that this method can be applied to selectively deliver plasmid vectors to the heart. METHODS AND RESULTS: We used albumin and lipid microbubbles containing plasmids with a luciferase transgene to target the heart in rats. After 4 days, organs were harvested and analyzed for reporter gene expression. In a second set of experiments, the hearts of rats treated with plasmids were harvested at various time points during a 4-week period. Both luciferase activity and mRNA concentrations were measured. Luciferase transfection with plasmids showed highly specific gene expression in the heart, with hardly any activity in control organs. Time course evaluation showed high transgene expression in the first 4 days, with a rapid decline thereafter. Repeated treatment produced a second peak of transgene expression with similar decay. CONCLUSIONS: Ultrasound-mediated destruction of microbubbles directs plasmid transgene expression to the heart with much greater specificity than viral vectors and can be regulated by repeated treatments. This noninvasive technique is a promising method for cardiac gene therapy. PMID- 12912824 TI - PseuRECA: genome annotation and gene context analysis for Pseudomonas aeruginosa PAO1. PMID- 12912822 TI - Serial cardiac magnetic resonance imaging of injected mesenchymal stem cells. AB - BACKGROUND: Delivery and tracking of endomyocardial stem cells are limited by the inability to image transplanted cells noninvasively in the beating heart. We hypothesized that mesenchymal stem cells (MSCs) could be labeled with a iron fluorophore particle (IFP) to provide MRI contrast in vivo to assess immediate and long-term localization. METHODS AND RESULTS: MSCs were isolated from swine. Short-term incubation of MSCs with IFP resulted in dose-dependent and efficient labeling. Labeled cells remained viable for multiple passages and retained in vitro proliferation and differentiation capacity. Labeled MSCs (10(4) to 10(6) cells/150 microL) were injected percutaneously into normal and freshly infarcted myocardium in swine. One, 3, and 1 animals underwent serial cardiac MRI (1.5T) for 4, 8, and 21 days, respectively. MRI contrast properties were measured both in vivo and in vitro for cells embedded in agar. Injection sites containing as few as 10(5) MSCs could be detected and contained intact IFP-bearing MSCs on histology. CONCLUSIONS: IFP labeling of MSCs imparts useful MRI contrast, enabling ready detection in the beating heart on a conventional cardiac MR scanner after transplantation into normal and infarcted myocardium. The dual labeled MSCs can be identified at locations corresponding to injection sites, both ex vivo using fluorescence microscopy and in vivo using susceptibility contrast on MRI. This technology may permit effective in vivo study of stem cell retention, engraftment, and migration. PMID- 12912825 TI - Comprehensive aligned sequence construction for automated design of effective probes (CASCADE-P) using 16S rDNA. AB - MOTIVATION: Prokaryotic organisms have been identified utilizing the sequence variation of the 16S rRNA gene. Variations steer the design of DNA probes for the detection of taxonomic groups or specific organisms. The long-term goal of our project is to create probe arrays capable of identifying 16S rDNA sequences in unknown samples. This necessitated the authentication, categorization and alignment of the >75 000 publicly available '16S' sequences. Preferably, the entire process should be computationally administrated so the aligned collection could periodically absorb 16S rDNA sequences from the public records. A complete multiple sequence alignment would provide a foundation for computational probe selection and facilitates microbial taxonomy and phylogeny. RESULTS: Here we report the alignment and similarity clustering of 62 662 16S rDNA sequences and an approach for designing effective probes for each cluster. A novel alignment compression algorithm, NAST (Nearest Alignment Space Termination), was designed to produce the uniform multiple sequence alignment referred to as the prokMSA. From the prokMSA, 9020 Operational Taxonomic Units (OTUs) were found based on transitive sequence similarities. An automated approach to probe design was straightforward using the prokMSA clustered into OTUs. As a test case, multiple probes were computationally picked for each of the 27 OTUs that were identified within the Staphylococcus Group. The probes were incorporated into a customized microarray and were able to correctly categorize Staphylococcus aureus and Bacillus anthracis into their correct OTUs. Although a successful probe picking strategy is outlined, the main focus of creating the prokMSA was to provide a comprehensive, categorized, updateable 16S rDNA collection useful as a foundation for any probe selection algorithm. PMID- 12912826 TI - A comparison of statistical methods for analysis of high density oligonucleotide array data. AB - MOTIVATION: Gene expression profiling has become an invaluable tool in functional genomics. A wide variety of statistical methods have been employed to analyze the data generated in experiments using Affymetrix GeneChip microarrays. It is important to understand the relative performance of these methods in terms of accuracy in detecting and quantifying relative gene expression levels and changes in gene expression. RESULTS: Three different analysis approaches have been compared in this work: non-parametric statistical methods implemented in Affymetrix Microarray Analysis Suite v5.0 (MAS5); an error-modeling based approach implemented in Rosetta Resolver v3.1; and an intensity-modeling approach implemented in dChip v1.1. A Latin Square data set generated and made available by Affymetrix was used in the comparison. All three methods-Resolver, MAS5 and the version of dChip based on the difference between perfect match and mismatch intensities-perform well in quantifying gene expression. Presence calls made by MAS5 and Resolver perform well at high concentrations, but they cannot be relied upon at low concentrations. The performance of Resolver and MAS5 in detecting 2 fold changes in transcript concentration is superior to that of dChip. At a comparable false positive rate, Resolver and MAS5 are able to detect many more true changes in transcript concentration. Estimated fold changes calculated by all the methods are biased below the true values. PMID- 12912827 TI - Differential expression in SAGE: accounting for normal between-library variation. AB - MOTIVATION: In contrasting levels of gene expression between groups of SAGE libraries, the libraries within each group are often combined and the counts for the tag of interest summed, and inference is made on the basis of these larger 'pseudolibraries'. While this captures the sampling variability inherent in the procedure, it fails to allow for normal variation in levels of the gene between individuals within the same group, and can consequently overstate the significance of the results. The effect is not slight: between-library variation can be hundreds of times the within-library variation. RESULTS: We introduce a beta-binomial sampling model that correctly incorporates both sources of variation. We show how to fit the parameters of this model, and introduce a test statistic for differential expression similar to a two-sample t-test. PMID- 12912828 TI - Class prediction and discovery using gene microarray and proteomics mass spectroscopy data: curses, caveats, cautions. AB - MOTIVATION: Two practical realities constrain the analysis of microarray data, mass spectra from proteomics, and biomedical infrared or magnetic resonance spectra. One is the 'curse of dimensionality': the number of features characterizing these data is in the thousands or tens of thousands. The other is the 'curse of dataset sparsity': the number of samples is limited. The consequences of these two curses are far-reaching when such data are used to classify the presence or absence of disease. RESULTS: Using very simple classifiers, we show for several publicly available microarray and proteomics datasets how these curses influence classification outcomes. In particular, even if the sample per feature ratio is increased to the recommended 5-10 by feature extraction/reduction methods, dataset sparsity can render any classification result statistically suspect. In addition, several 'optimal' feature sets are typically identifiable for sparse datasets, all producing perfect classification results, both for the training and independent validation sets. This non uniqueness leads to interpretational difficulties and casts doubt on the biological relevance of any of these 'optimal' feature sets. We suggest an approach to assess the relative quality of apparently equally good classifiers. PMID- 12912829 TI - Serial BLAST searching. AB - MOTIVATION: The translating BLAST algorithms are powerful tools for finding protein-coding genes because they identify amino acid similarities in nucleotide sequences. Unfortunately, these kinds of searches are computationally intensive and often represent bottlenecks in sequence analysis pipelines. Tuning parameters for speed can make the searches much faster, but one risks losing low-scoring alignments. However, high scoring alignments are relatively resistant to such changes in parameters, and this fact makes it possible to use a serial strategy where a fast, insensitive search is used to pre-screen a database for similar sequences, and a slow, sensitive search is used to produce the sequence alignments. RESULTS: Serial BLAST searches improve both the speed and sensitivity. PMID- 12912830 TI - Greedy method for inferring tandem duplication history. AB - MOTIVATION: Genome analysis suggests that tandem duplication is an important mode of evolutionary novelty by permitting one copy of each gene to drift and potentially to acquire a new function. With more and more genomic sequences available, reconstructing duplication history has received extensive attention recently. RESULTS: An efficient method is presented for inferring the duplication history of tandemly repeated sequences based on the model proposed by Fitch (1977). We validate the method by using simulation results and real data sets of mucin genes, ZNF genes, and olfactory receptors genes. The agreement with conclusions drawn by other biological researchers strongly indicates that our method is efficient and robust. AVAILABILITY: The program is available by request. PMID- 12912831 TI - A hidden Markov model for progressive multiple alignment. AB - MOTIVATION: Progressive algorithms are widely used heuristics for the production of alignments among multiple nucleic-acid or protein sequences. Probabilistic approaches providing measures of global and/or local reliability of individual solutions would constitute valuable developments. RESULTS: We present here a new method for multiple sequence alignment that combines an HMM approach, a progressive alignment algorithm, and a probabilistic evolution model describing the character substitution process. Our method works by iterating pairwise alignments according to a guide tree and defining each ancestral sequence from the pairwise alignment of its child nodes, thus, progressively constructing a multiple alignment. Our method allows for the computation of each column minimum posterior probability and we show that this value correlates with the correctness of the result, hence, providing an efficient mean by which unreliably aligned columns can be filtered out from a multiple alignment. PMID- 12912832 TI - Molecular evaluation using in silico protein interaction profiles. AB - MOTIVATION: To find a correlation between the activities and structures of molecules is one of the most important subjects for molecular evaluation study. Traditional quantitative structure-activity relationship (QSAR) methodologies represent those attempts using physicochemical descriptors. Creating a new molecular description factor based on the results of a computational docking study will add new dimensions to molecular evaluation. RESULTS: We propose a new molecular description factor analysis system called the Comparative Molecular Interaction Profile Analysis (CoMIPA) system in which the AutoDock program is used for docking evaluation of small molecule compound-protein complexes. Interaction energies are calculated, and the data sets obtained are called interaction profiles (IPFs). Using the IPF as a scoring indicator, the system could be a powerful tool to cluster the interacting properties between small molecules and bio macromolecules such as ligand-receptor bindings. Further development of the system will enable us to predict the adverse effects of a drug candidate. PMID- 12912833 TI - Identification of functional links between genes using phylogenetic profiles. AB - MOTIVATION: Genes with identical patterns of occurrence across the phyla tend to function together in the same protein complexes or participate in the same biochemical pathway. However, the requirement that the profiles be identical (i) severely restricts the number of functional links that can be established by such phylogenetic profiling; (ii) limits detection to very strong functional links, failing to capture relations between genes that are not in the same pathway, but nevertheless subserve a common function and (iii) misses relations between analogous genes. Here we present and apply a method for relaxing the restriction, based on the probability that a given arbitrary degree of similarity between two profiles would occur by chance, with no biological pressure. Function is then inferred at any desired level of confidence. RESULTS: We derive an expression for the probability distribution of a given number of chance co-occurrences of a pair of non-homologous orthologs across a set of genomes. The method is applied to 2905 clusters of orthologous genes (COGs) from 44 fully sequenced microbial genomes representing all three domains of life. Among the results are the following. (1) Of the 51 000 annotated intrapathway gene pairs, 8935 are linked at a level of significance of 0.01. This is over 30-fold greater than the 271 intrapathway pairs obtained at the same confidence level when identical profiles are used. (2) Of the 540 000 interpathway genes pairs, some 65 000 are linked at the 0.01 level of significance, some 12 standard deviations beyond the number expected by chance at this confidence level. We speculate that many of these links involve nearest-neighbor path, and discuss some examples. (3) The difference in the percentage of linked interpathway and intrapathway genes is highly significant, consistent with the intuitive expectation that genes in the same pathway are generally under greater selective pressure than those that are not. (4) The method appears to recover well metabolic networks. This is illustrated by the TCA cycle which is recovered as a highly connected, weighted edge network of 30 of its 31 COGs. (5) The fraction of pairs having a common pathway is a symmetric function of the Hamming distance between their profiles. This finding, that the functional correlation between profiles with near maximum Hamming distance is as large as between profiles with near zero Hamming distance, and as statistically significant, is plausibly explained if the former group represents analogous genes. PMID- 12912834 TI - Probabilistic scoring measures for profile-profile comparison yield more accurate short seed alignments. AB - MOTIVATION: The development of powerful automatic methods for the comparison of protein sequences has become increasingly important. Profile-to-profile comparisons allow for the use of broader information about protein families, resulting in more sensitive and accurate comparisons of distantly related sequences. A key part in the comparison of two profiles is the method for the calculation of scores for the position matches. A number of methods based on various theoretical considerations have been proposed. We implemented several previously reported scoring functions as well as our own functions, and compared them on the basis of their ability to produce accurate short ungapped alignments of a given length. RESULTS: Our results suggest that the family of the probabilistic methods (log-odds based methods and prof_sim) may be the more appropriate choice for the generation of initial 'seeds' as the first step to produce local profile-profile alignments. The most effective scoring systems were the closely related modifications of functions previously implemented in the COMPASS and Picasso methods. PMID- 12912835 TI - Development of a four-body statistical pseudo-potential to discriminate native from non-native protein conformations. AB - MOTIVATION: Most scoring functions used in protein fold recognition employ two body (pseudo) potential energies. The use of higher-order terms may improve the performance of current algorithms. METHODS: Proteins are represented by the side chain centroids of amino acids. Delaunay tessellation of this representation defines all sets of nearest neighbor quadruplets of amino acids. Four-body contact scoring function (log likelihoods of residue quadruplet compositions) is derived by the analysis of a diverse set of proteins with known structures. A test protein is characterized by the total score calculated as the sum of the individual log likelihoods of composing amino acid quadruplets. RESULTS: The scoring function distinguishes native from partially unfolded or deliberately misfolded structures. It also discriminates between pre- and post-transition state and native structures in the folding simulations trajectory of Chymotrypsin Inhibitor 2 (CI2). PMID- 12912836 TI - Rapid classification of a protein fold family using a statistical analysis of dipolar couplings. AB - MOTIVATION: One of the primary aims of the structural genomics initiative is the determination of representative structures from each protein fold family. Given this objective, it is important to rapidly identify proteins that belong to a family that is already well populated (so they can be eliminated from further studies), or more importantly identify proteins that represent new families of fold. RESULTS: A method for rapid classification to a fold family by the statistical analyses of unassigned the (15)N-(1)H residual dipolar couplings is presented. The required NMR data can be quickly acquired and analyzed. Using this method, structure determination efforts can be focused on more unique and interesting structures, and the overall efficiency in the construction of an information-rich library can be increased. PMID- 12912837 TI - Pandit: a database of protein and associated nucleotide domains with inferred trees. AB - MOTIVATION: A large, high-quality database of homologous sequence alignments with good estimates of their corresponding phylogenetic trees will be a valuable resource to those studying phylogenetics. It will allow researchers to compare current and new models of sequence evolution across a large variety of sequences. The large quantity of data may provide inspiration for new models and methodology to study sequence evolution and may allow general statements about the relative effect of different molecular processes on evolution. RESULTS: The Pandit 7.6 database contains 4341 families of sequences derived from the seed alignments of the Pfam database of amino acid alignments of families of homologous protein domains (Bateman et al., 2002). Each family in Pandit includes an alignment of amino acid sequences that matches the corresponding Pfam family seed alignment, an alignment of DNA sequences that contain the coding sequence of the Pfam alignment when they can be recovered (overall, 82.9% of sequences taken from Pfam) and the alignment of amino acid sequences restricted to only those sequences for which a DNA sequence could be recovered. Each of the alignments has an estimate of the phylogenetic tree associated with it. The tree topologies were obtained using the neighbor joining method based on maximum likelihood estimates of the evolutionary distances, with branch lengths then calculated using a standard maximum likelihood approach. PMID- 12912838 TI - Evaluation of ontology development tools for bioinformatics. AB - Ontologies are being used nowadays in many areas, including bioinformatics. To assist users in developing and maintaining ontologies a number of tools have been developed. In this paper we compare four such tools, Protege-2000, Chimaera, DAG Edit and OilEd. As test ontologies we have used ontologies from the Gene Ontology Consortium. No system is preferred in all situations, but each system has its own strengths and weaknesses. PMID- 12912839 TI - MrBayes 3: Bayesian phylogenetic inference under mixed models. AB - MrBayes 3 performs Bayesian phylogenetic analysis combining information from different data partitions or subsets evolving under different stochastic evolutionary models. This allows the user to analyze heterogeneous data sets consisting of different data types-e.g. morphological, nucleotide, and protein and to explore a wide variety of structured models mixing partition-unique and shared parameters. The program employs MPI to parallelize Metropolis coupling on Macintosh or UNIX clusters. PMID- 12912840 TI - AGenDA: homology-based gene prediction. AB - We present a www server for homology-based gene prediction. The user enters a pair of evolutionary related genomic sequences, for example from human and mouse. Our software system uses CHAOS and DIALIGN to calculate an alignment of the input sequences and then searches for conserved splicing signals and start/stop codons around regions of local sequence similarity. This way, candidate exons are identified that are used, in turn, to calculate optimal gene models. The server returns the constructed gene model by email, together with a graphical representation of the underlying genomic alignment. PMID- 12912841 TI - RankGene: identification of diagnostic genes based on expression data. AB - RankGene is a program for analyzing gene expression data and computing diagnostic genes based on their predictive power in distinguishing between different types of samples. The program integrates into one system a variety of popular ranking criteria, ranging from the traditional t-statistic to one-dimensional support vector machines. This flexibility makes RankGene a useful tool in gene expression analysis and feature selection. PMID- 12912842 TI - UniFrag and GenomePrimer: selection of primers for genome-wide production of unique amplicons. AB - The complementary programs UniFrag and GenomePrimer were developed to provide a reliable high-throughput method to select the most unique regions within genomic DNA sequence(s) and design primers therein, involving minimal user intervention and maximum flexibility. PMID- 12912843 TI - GenoMap, a circular genome data viewer. AB - A Tcl/Tk-based application called GenoMap is described, a viewer for genome-wide map of microarray expression data within a circular bacterial genome. An interactive interface facilitates easy identification of the expressed region. This software is also used for drawing genome-wide quantitative data. PMID- 12912844 TI - ClustalW-MPI: ClustalW analysis using distributed and parallel computing. AB - ClustalW is a tool for aligning multiple protein or nucleotide sequences. The alignment is achieved via three steps: pairwise alignment, guide-tree generation and progressive alignment. ClustalW-MPI is a distributed and parallel implementation of ClustalW. All three steps have been parallelized to reduce the execution time. The software uses a message-passing library called MPI (Message Passing Interface) and runs on distributed workstation clusters as well as on traditional parallel computers. PMID- 12912845 TI - ESTWeb: bioinformatics services for EST sequencing projects. AB - ESTWeb is an internet based software package designed for uniform data processing and storage for large-scale EST sequencing projects. The package provides for: (a) reception of sequencing chromatograms; (b) sequence processing such as base calling, vector screening, comparison with public databases; (c) storage of data and analysis in a relational database, (d) generation of a graphical report of individual sequence quality; and (e) issuing of reports with statistics of productivity and redundancy. The software facilitates real-time monitoring and evaluation of EST sequence acquisition progress along an EST sequencing project. PMID- 12912846 TI - PISCES: a protein sequence culling server. AB - PISCES is a public server for culling sets of protein sequences from the Protein Data Bank (PDB) by sequence identity and structural quality criteria. PISCES can provide lists culled from the entire PDB or from lists of PDB entries or chains provided by the user. The sequence identities are obtained from PSI-BLAST alignments with position-specific substitution matrices derived from the non redundant protein sequence database. PISCES therefore provides better lists than servers that use BLAST, which is unable to identify many relationships below 40% sequence identity and often overestimates sequence identity by aligning only well conserved fragments. PDB sequences are updated weekly. PISCES can also cull non PDB sequences provided by the user as a list of GenBank identifiers, a FASTA format file, or BLAST/PSI-BLAST output. PMID- 12912847 TI - QAlign: quality-based multiple alignments with dynamic phylogenetic analysis. AB - Integrating different alignment strategies, a layout editor and tools deriving phylogenetic trees in a 'multiple alignment environment' helps to investigate and enhance results of multiple sequence alignment by hand. QAlign combines algorithms for fast progressive and accurate simultaneous multiple alignment with a versatile editor and a dynamic phylogenetic analysis in a convenient graphical user interface. PMID- 12912848 TI - Digital extractor: analysis of digital differential display output. AB - Digital Extractor is a program for the high-throughput processing of data sets derived from digital differential display-based comparisons of EST libraries. These comparisons can be utilized to identify discrete subsets of genes whose expression is restricted to distinct tissue types. The program facilitates these investigations by permitting parallel annotation of genes identified as being differentially expressed. PMID- 12912849 TI - Capsaicin and menthol in the treatment of itch and pain: recently cloned receptors provide the key. AB - Topical capsaicin is reported to be an effective treatment for idiopathic intractable pruritus ani. While both capsaicin and menthol application produce a transient perianal burning sensation, only capsaicin relieves itching. Classical observations on functional desensitisation of nociceptors by capsaicin may explain the beneficial effects but the recent discovery of a range of receptors which respond to capsaicin, menthol, and temperature, and their expression in subsets of sensory nerve fibres, provides an exciting prospect towards advancing our understanding and treatment of sensory dysfunction. PMID- 12912850 TI - In vivo electron spin resonance spectroscopy: what use is it to gastroenterologists? PMID- 12912851 TI - Keeping neuroendocrine cells in check: roles for TGFbeta, Smads, and menin? PMID- 12912852 TI - Inappropriate ileal conservation of bile acids in cholestatic liver disease: homeostasis gone awry. PMID- 12912853 TI - Endogenous prostaglandin I2 regulates the neural emergency system through release of calcitonin gene related peptide. AB - BACKGROUND: We previously reported that endogenous prostaglandin I(2), generated by a mild irritant, sensitised calcitonin gene related peptide (CGRP) containing sensory nerves and facilitated the release of CGRP and gastric mucosal protection against ethanol. Administration of capsaicin also inhibited ethanol induced gastric mucosal injury through immediate release of CGRP from primary sensory neurones, which is termed the neural emergency system. In the present study, we tested whether endogenous prostaglandin I(2) also modulates the cytoprotective action of capsaicin using prostaglandin I receptor knockout mice (IP(-/-)). METHODS: The stomachs of IP(-/-) or their wild-type counterparts (IP(+/+)), anaesthetised with urethane (1.225 g/kg), were doubly cannulated from the oesophageal and duodenal sides, and the gastric mucosa was perfused (1 ml/min) with physiological saline. Perfusate was changed to 50% ethanol alone, or 50% ethanol containing capsaicin (16 approximately 1600 micro M). The injured area was estimated at the end of each perfusion experiment. In some animals, CGRP-(8 37), a CGRP antagonist (0.3 mg/kg), or indomethacin (1 mg/kg) was intravenously injected before perfusion of 50% ethanol containing capsaicin. RESULTS: Capsaicin inhibited the injured area in a dose dependent manner. Fifty per cent ethanol containing capsaicin (480 micro M) immediately increased intragastric levels of CGRP although 50% ethanol alone did not. The protective action of capsaicin (480 micro M) against ethanol was completely abolished by intravenous injection of CGRP-(8-37). Indomethacin also inhibited the protective action of capsaicin, and this was accompanied by reduced levels of intragastric CGRP. Intragastric levels of prostaglandin E(2) were not increased by capsaicin treatment but those of 6 keto-prostaglandin F(1alpha), a metabolite of prostaglandin I(2), were markedly increased. No protective action of capsaicin was observed in IP(-/-) which lacked the ability to increase intragastric CGRP levels in response to ethanol containing capsaicin. The CGRP content of the stomach from untreated IP(-/-) did not differ from those in IP(+/+). Capsaicin (160 micro M) together with intragastric perfusion of beraprost sodium (PGI(2) analogue, 2.5 micro g/ml) showed enhanced protection against ethanol induced injury. This enhanced protection was completely blocked by intravenous injection of CGRP-(8-37). CONCLUSIONS: The present results suggest that endogenous prostaglandin I(2) enhances the protective action of the capsaicin mediated neural emergency system against ethanol induced gastric mucosal injury through enhancement of CGRP release. PMID- 12912854 TI - Interleukin 1beta induces gastric epithelial cell matrix metalloproteinase secretion and activation during Helicobacter pylori infection. AB - BACKGROUND: and aims: Matrix metalloproteinases (MMPs) are endopeptidases with roles in extracellular matrix remodelling, cell proliferation, and inflammatory processes. We showed previously that Helicobacter pylori infection of human gastric adenocarcinoma (AGS) cells increased epithelial secretion of epithelial MMP-1 and MMP-3 and bacterial secretion of MMP-3-like activity. In the present study, we sought to characterise the role of interleukin (IL)-1beta in H pylori induced secretion of epithelial MMPs. METHODS AND RESULTS: AGS cells were treated with H pylori and/or IL-1beta. Comparable IL-8 secretory responses (approximately 1700 ng/ml) measured by ELISA were induced by 2.0 ng/ml IL-1beta and by H pylori at a multiplicity of infection (MOI) of 50. The same IL-1beta and H pylori concentrations induced comparable increases in AGS cell caseinolytic activity at 60 kDa. MMP-3 monoclonal antibody immunoblots of AGS cell conditioned media detected immunoreactive bands at 71 kDa and 56 kDa. H pylori (MOI=50-100) induced dose dependent increases in both bands whereas IL-1beta (0.2-2 ng/ml) induced dose dependent increases only in the 71 kDa band, which was identified as a MMP 3/TIMP-3 (tissue inhibitor of metalloproteinases 3) heterodimer. AGS/H pylori conditioned media expressed 24 times more MMP-3 activity than AGS/IL-1beta conditioned media. There was a strong interaction between IL-1beta and H pylori on MMP-3 secretion. CONCLUSIONS: We conclude that IL-1beta induces gastric epithelial cell MMP-3 secretion, contributing to epithelial tissue destruction during H pylori infection. However, other bacterial/host factors are needed to mediate the full gastric epithelial cell MMP-3 secretory response induced by H pylori infection. PMID- 12912855 TI - Monocyte chemoattractant protein 1 (MCP-1) released from Helicobacter pylori stimulated gastric epithelial cells induces cyclooxygenase 2 expression and activation in T cells. AB - BACKGROUND: and aims: To clarify the interaction between gastric epithelial and mucosal T cells, we examined the role of cytokines released from epithelial cells in response to Helicobacter pylori water extract protein (HPWEP) in regulating T cell cyclooxygenase 2 (COX-2) expression and activation. METHODS: Media from MKN 28 cells incubated with HPWEP for 48 hours were added to Jurkat T cells and human peripheral T cells. C-C and CXC chemokine concentrations in MKN-28 cell media, and COX-2 expression, interferon gamma (IFN-gamma), and interleukin (IL)-4 secretions in T cells were determined by western blot analysis and ELISA methods. Distributions of COX-2 positive T cells and monocyte chemoattractant protein 1 (MCP-1) in tissue specimens with H pylori associated gastritis were determined as single or double labelling by immunohistochemistry. RESULTS: MCP-1, IL-7, IL-8, and RANTES were detected in media from MKN-28 cells incubated with HPWEP. Media as a whole, and MCP-1 alone, stimulated COX-2 expression and peripheral T cell proliferation. Anti-MCP-1 antibody inhibited media stimulated COX-2 mRNA expression in Jurkat T cells. Media stimulated IFN-gamma but not IL-4 secretion from peripheral T cells, while MCP-1 stimulated IL-4 but not IFN-gamma secretion. Both stimulated cytokine release, and peripheral T cell proliferation was partially inhibited by NS-398, a specific COX-2 inhibitor. In mucosa with gastritis, COX-2 was expressed in T cells and MCP-1 was localised mainly in epithelial and mononuclear cells. MCP-1 levels and the intensity of COX-2 expression in tissue samples were closely related. CONCLUSIONS: Cytokines such as MCP-1, released from gastric epithelial cells in response to HPWEP, seem to modulate T cell immune responses, at least in part via COX-2 expression. PMID- 12912856 TI - Channelling bias and the incidence of gastrointestinal haemorrhage in users of meloxicam, coxibs, and older, non-specific non-steroidal anti-inflammatory drugs. AB - BACKGROUND: Although clinical trial results suggest that meloxicam has less gastrointestinal toxicity than most other non-steroidal anti-inflammatory drugs (NSAIDs), in practice it has been associated with a large number of yellow card reports of gastrointestinal complications. AIMS: To estimate whether meloxicam and the coxibs, rofecoxib and celecoxib, have been channelled towards high risk patients, and to estimate the risk of hospitalisation for gastrointestinal haemorrhage associated with the use of these drugs, allowing for the effects of channelling. PATIENTS: Using the UK General Practice Research Database, this study included 7.1 thousand patient years (tpy) exposure to meloxicam, 1.6 tpy exposure to coxibs, and 628 tpy exposure to older non-specific NSAIDs. METHODS: Cohort study of patients who received a prescription for an NSAID between June 1987 and January 2001. Exposure to newer NSAIDs (meloxicam, rofecoxib, celecoxib) and to older non-specific NSAIDs was identified. Channelling was assessed on factors including: demographic variables; diagnosis of arthritis; history of NSAID use or gastrointestinal events, including gastrointestinal haemorrhage; and use of ulcer healing drugs. RESULTS: Most risk factors for gastrointestinal haemorrhage were more prevalent among patients prescribed the newer NSAIDs. Adjusting for these risk factors reduced the relative risks of gastrointestinal haemorrhage on meloxicam and coxibs versus older non-specific NSAIDs to 0.84 (95% confidence interval 0.60, 1.17) and 0.36 (0.14, 0.97), respectively. CONCLUSIONS: Channelling towards high risk gastrointestinal patients occurred in the prescribing of newer NSAIDs. After attempting to correct for channelling bias, coxib exposure, but not meloxicam exposure, was associated with a significantly lower risk of gastrointestinal haemorrhage than older non-specific NSAID exposure. PMID- 12912857 TI - Assessment of meal induced gastric accommodation by a satiety drinking test in health and in severe functional dyspepsia. AB - AIMS: Impaired gastric accommodation is a major pathophysiological mechanism in functional dyspepsia. The aim of the present work was to assess a satiety drinking test in the evaluation of accommodation in health and dyspepsia. METHODS: Twenty five controls and 37 severely dyspeptic patients seen at a tertiary care centre completed a dyspepsia questionnaire, and gastric emptying and gastric barostat studies. The amount of liquid meal ingested at maximum satiety during a slow satiety drinking test was determined. In controls, we studied the influence of caloric density and of pharmacological agents that influence accommodation. RESULTS: In patients, satiety scores were higher and maximum satiety occurred at lower calories (542 (50) v 1508 (53) kcal; p<0.0001). Six patients had required nutritional support, but excluding these did not alter the correlations. With increasing severity of early satiety, less calories were ingested at maximum satiety. In multivariate analysis, the amount of calories was significantly correlated to accommodation but not to gastric emptying or sensitivity. Sensitivity and specificity of the satiety test in predicting impaired accommodation reached 92% and 86%, respectively. At different caloric densities, ingested volume rather than caloric load determined maximum satiety. Pharmacological agents (sumatriptan, cisapride, erythromycin) affected the satiety test according to their effect on accommodation. CONCLUSION: A slow caloric drinking test can be used to evaluate accommodation and early satiety. It provides a non-invasive method of predicting impaired accommodation and quantifying pharmacological influences on accommodation. PMID- 12912858 TI - Immunomodulatory effects of inhaled carbon monoxide on rat syngeneic small bowel graft motility. AB - BACKGROUND: Intestinal transplantation provokes an intense inflammatory response within the graft muscularis that causes intestinal ileus. We hypothesised that endogenously produced anti-inflammatory substances could be utilised as novel therapeutics. Therefore, we tested the protective effects of inhaled carbon monoxide (CO) and an endogenous haeme oxygenase 1 (HO-1) anti-inflammatory mediator on transplant induced inflammatory responses and intestinal ileus in the rat. METHODS: Gastrointestinal transit of non-absorbable FITC labelled dextran and in vitro jejunal circular muscle contractions were measured in controls and syngeneic orthotopic transplanted animals with and without CO inhalation (250 ppm for 25 hours). Inflammatory mRNAs for interleukin (IL)-6, IL-1beta, tumour necrosis factor alpha (TNF-alpha), intercellular adhesion molecule 1 (ICAM-1), inducible nitric oxide (iNOS), cyclooxygenase 2 (COX-2), and IL-10 were quantified by real time reverse transcriptase-polymerase chain reaction and HO-1 by northern blot. Histochemical stains characterised neutrophil infiltration and enterocyte apoptosis. RESULTS: Transplantation delayed transit and suppressed jejunal circular muscle contractility. Transplantation induced dysmotility was significantly improved by CO inhalation. Transplantation initiated a significant upregulation in IL-6, IL-1beta, TNF-alpha, ICAM-1, iNOS, COX-2, and HO-1 mRNAs with the graft muscularis. CO inhalation significantly decreased expression of IL 6, IL-1beta, iNOS, and COX-2 mRNAs. CO also significantly decreased serum nitrite levels (iNOS activity). CONCLUSIONS: CO inhalation significantly improved post transplant motility and attenuated the inflammatory cytokine milieu in the syngeneic rat transplant model. Thus clinically providing CO, the end product of the anti-inflammatory HO-1 pathway, may prove to be an effective therapeutic adjunct for clinical small bowel transplantation. PMID- 12912859 TI - Interferon beta-1a in ulcerative colitis: a placebo controlled, randomised, dose escalating study. AB - BACKGROUND: and aims: Administration of interferon (IFN)-beta may represent a rational approach to the treatment of ulcerative colitis through its immunomodulatory and anti-inflammatory effects. The present study was performed to evaluate the efficacy and tolerability of IFN-beta-1a. METHODS: Patients (n=18) with moderately active ulcerative colitis were randomised to receive IFN beta-1a or placebo. IFN-beta-1a was started at a dose of 22 micro g three times a week subcutaneously, and the dose was increased at two week intervals to 44 micro g and then to 88 micro g if no response was observed. The maximum duration of treatment was eight weeks. End points were clinical treatment response, defined as a decrease of at least 3 points from baseline in the ulcerative colitis scoring system (UCSS) symptoms score and induction of endoscopically confirmed remission. RESULTS: Baseline characteristics and disease severity were similar in both groups. Data from 17 patients are included in this report (10 patients in the IFN-beta-1a group and seven patients in the placebo group). Clinical response was achieved in five patients (50%) in the IFN-beta-1a group and in one (14%) in the placebo group (P=0.14). Remission was achieved in three patients in the IFN beta-1a group and in none in the placebo group (p=0.02). Most adverse reactions associated with IFN-beta-1a were influenza-like symptoms or injection site reactions, and were mild or moderate in severity. CONCLUSIONS: IFN-beta-1a may represent a promising novel treatment approach in ulcerative colitis. PMID- 12912860 TI - Mucosal sulfhydryl compounds evaluation by in vivo electron spin resonance spectroscopy in mice with experimental colitis. AB - BACKGROUND: Sulfhydryl (SH) compounds are essential in maintaining mucosal integrity in the gastrointestinal tract. A decrease in colonic mucosal SH compounds affects the redox status of the mucosa, resulting in vulnerability to further attacks. Therefore, there is a strong need for in vivo evaluation of SH compounds in the colonic mucosa. AIMS: The aim of the current study was to establish a method of evaluating levels of SH compounds in the colonic mucosa of live animals before and after induction of colitis. METHODS: Murine experimental colitis was induced by instillation of trinitrobenzene sulphonic acid (TNBS) dissolved in 50% ethanol into the colon via the anus. For evaluation of mucosal SH compounds in the colon, 3-carbamoyl-2,2,5,5-tetramethylpyrrolidine-1-oxyl (carbamoyl-PROXYL), a stable nitroxide radical, was instilled into the colonic lumen of live mice and the spin clearance rate was measured by L-band electron spin resonance (ESR) spectroscopy. RESULTS: Morphological study showed that mucosal damage was severe one or two days after TNBS instillation. The colonic mucosa started to regenerate at four days, and looked normal at seven days, after induction of colitis. The spin clearance rate of carbamoyl-PROXYL decreased significantly at 0.5, 1, 2, and 4 days after induction of colitis compared with mice before TNBS instillation. Surprisingly, although the colonic mucosa looked normal seven days after TNBS administration, the spin clearance rate still remained significantly slow. The spin clearance rate returned to normal 14 days after induction of colitis. The change in in vivo spin clearance rate was consistent with the time dependent change in mucosal reduced glutathione, a major component of SH compounds. CONCLUSION: The spin clearance rate obtained by L-band ESR spectroscopy in combination with carbamoyl-PROXYL can give an estimate of the level of colonic mucosal SH compounds in live animals and is useful for evaluating the mucosal defence system against oxidative stress. PMID- 12912861 TI - Tumour necrosis factor alpha and nuclear factor kappaB inhibit transcription of human TFF3 encoding a gastrointestinal healing peptide. AB - BACKGROUND: and aims: Tumour necrosis factor alpha (TNF-alpha) induction of nuclear factor kappaB (NFkappaB) activation plays a major role in the pathogenesis of inflammatory bowel disease (IBD). Trefoil factor family peptides TFF1, TFF2, and TFF3 exert protective, curative, and tumour suppressive functions in the gastrointestinal tract. In this study, we investigated effects of the TNF alpha/NFkappaB regulatory pathway by TNF-alpha on expression of TFFs. METHODS: After TNF-alpha stimulation, expression of TFF genes was analysed by quantitative real time polymerase chain reaction and by reporter gene assays in the gastrointestinal tumour cell lines HT-29 and KATO III. Additionally, NFkappaB subunits and a constitutive repressive form of inhibitory factor kappaB (IkappaB) were transiently coexpressed. In vivo, morphological changes and expression of TFF3, mucins, and NFkappaB were monitored by immunohistochemistry in a rat model of 2,4,6-trinitrobenzene sulphonic acid induced colitis. RESULTS: TNF-alpha stimulation evoked up to 10-fold reduction of TFF3 expression in the colon tumour cell line HT-29. Downregulation of reporter gene transcription of TFF3 was observed with both TNF-alpha and NFkappaB, and was reversible by IkappaB. In vivo, the increase in epithelial expression of NFkappaB coincided with reduced TFF3 expression during the acute phase of experimental colitis. CONCLUSIONS: Downregulation of intestinal trefoil factor TFF3 is caused by repression of transcription through TNF-alpha and NFkappaB activation in vitro. In IBD, perpetual activation of NFkappaB activity may contribute to ulceration and decreased wound healing through reduced TFF3. PMID- 12912862 TI - Telomere shortening of epithelial cells characterises the adenoma-carcinoma transition of human colorectal cancer. AB - BACKGROUND: and aims: Chromosomal instability is one of the most consistent markers of sporadic colorectal cancer in humans. There is growing evidence that telomere shortening is one of the mechanisms leading to chromosomal instability and cancer initiation. METHODS: To test this hypothesis, the telomere length of colorectal epithelial cells and cells from connective tissue was determined at the adenoma-carcinoma transition at the cellular level by quantitative fluorescence in situ hybridisation. RESULTS: Our study showed that the telomere fluorescence intensity of epithelial cells was significantly weaker at the earliest morphologically definable stage of carcinoma-high grade dysplasia with minimal invasive growth-compared with the surrounding adenoma. In contrast, cells from connective tissue had a similar telomere signal intensity at the carcinoma stage compared with the adenoma, and in turn cells from connective tissue had overall significantly stronger telomere fluorescence signals compared with epithelial cells. CONCLUSIONS: These results demonstrate that short telomeres of epithelial cells characterise the adenoma-carcinoma transition during human colorectal carcinogenesis, suggesting that carcinomas arise from cells with critical short telomeres within the adenoma. Since the adenoma-carcinoma transition in colorectal cancer is characterised by an increase in chromosomal instability and anaphase bridges, our data support the hypothesis that short telomeres initiate colorectal cancer by induction of chromosomal instability. PMID- 12912863 TI - Autocrine growth inhibition by transforming growth factor beta-1 (TGFbeta-1) in human neuroendocrine tumour cells. AB - BACKGROUND AND AIM: The role of transforming growth factor beta-1 (TGFbeta-1) in neuroendocrine tumour biology is currently unknown. We therefore examined the expression and biological significance of TGFbeta signalling components in neuroendocrine tumours (NETs) of the gastroenteropancreatic (GEP) tract. METHODS: Expression of TGFbeta-1 and its receptors, Smads and Smad regulated proteins, was examined in surgically resected NET specimens and human NET cell lines by immunohistochemistry, reverse transcriptase-polymerase chain reaction, immunoblotting, and ELISA. Activation of TGFbeta-1 dependent promoters was tested by transactivation assays. Growth regulation was evaluated by cell numbers, soft agar assays, and cell cycle analysis using flow cytometry. The role of endogenous TGFbeta was assessed by a TGFbeta neutralising antibody and stable transfection of a dominant negative TGFbetaR II receptor construct. RESULTS: Coexpression of TGFbeta-1 and its receptors TGFbetaR I and TGFbetaR II was detected in 67% of human NETs and in all three NET cell lines examined. NET cell lines expressed the TGFbeta signal transducers Smad 2, 3, and 4. In two of the three cell lines, TGFbeta-1 treatment resulted in transactivation of a TGFbeta responsive reporter construct as well as inhibition of c-myc and induction of p21((WAF1)) expression. TGFbeta-1 inhibited anchorage dependent and independent growth in a time and dose dependent manner in TGFbeta-1 responsive cell lines. TGFbeta-1 mediated growth inhibition was due to G1 arrest without evidence of induction of apoptosis. Functional inactivation of endogenous TGFbeta revealed the existence of an autocrine antiproliferative loop in NET cells. CONCLUSIONS: Neuroendocrine tumour cells of the gastroenteropancreatic tract are subject to paracrine and autocrine growth inhibition by TGFbeta-1, which may account in part for the low proliferative index of this tumour entity. PMID- 12912864 TI - Potential role for peroxisome proliferator activated receptor (PPAR) in preventing colon cancer. AB - BACKGROUND: Peroxisome proliferator activated receptors (PPARs) are nuclear hormone receptors involved in genetic control of many cellular processes. PPAR and PPAR have been implicated in colonic malignancy. Here we provide three lines of evidence suggesting an inhibitory role for PPAR in colorectal cancer development. METHODS: Levels of PPAR mRNA and protein in human colorectal cancers were compared with matched non-malignant mucosa using RNAse protection and western blotting. APC(Min)/+ mice were randomised to receive the PPAR activator methylclofenapate 25 mg/kg or vehicle for up to 16 weeks, and small and large intestinal polyps were quantified by image analysis. The effect of methylclofenapate on serum stimulated mitogenesis (thymidine incorporation), linear cell growth, and annexin V and propidium iodide staining were assessed in human colonic epithelial cells. RESULTS: PPAR (mRNA and protein) expression levels were significantly depressed in colorectal cancer compared with matched non-malignant tissue. Methylclofenapate reduced polyp area in the small intestine from 18.7 mm(2) (median (interquartile range 11.1, 26.8)) to 9.90 (4.88, 13.21) mm(2) (p=0.003) and in the colon from 9.15 (6.31, 10.5) mm(2) to 3.71 (2.71, 5.99) mm(2) (p=0.009). Methylclofenapate significantly reduced thymidine incorporation and linear cell growth with no effect on annexin V or propidium iodide staining. CONCLUSIONS: PPAR may inhibit colorectal tumour progression, possibly via inhibition of proliferation, and may be an important therapeutic target. PMID- 12912865 TI - Topical capsaicin--a novel and effective treatment for idiopathic intractable pruritus ani: a randomised, placebo controlled, crossover study. AB - PURPOSE: Pruritus ani is a common and embarrassing proctological condition which can be very difficult to treat. We report the results of a double blind placebo controlled study of treatment with capsaicin. METHODS: Firstly, a pilot open study was carried out on five patients to establish which of two doses was the most acceptable by comparing effectiveness and side effects. Secondly, a double blind, placebo controlled, crossover study of topical capsaicin was performed. This study involved two four week treatment phases separated by a one week washout phase. Forty four patients were randomised to receive locally either active capsaicin (0.006%) or placebo (menthol 1%) ointment over a four week period (22 patients per group). After four weeks of treatment and a one week washout period, the placebo group began to receive capsaicin while the treated group received placebo (menthol 1%) for another four weeks. At the end of the controlled study, responders from both groups continued with capsaicin treatment in an open labelled manner. RESULTS: Thirty one of 44 patients experienced relief during capsaicin treatment periods and did not respond to menthol; all patients not responding to capsaicin also failed on menthol (p<0.0001). In 13 patients, treatment with capsaicin was unsuccessful: eight patients did not respond to capsaicin treatment, one responded equally to capsaicin and placebo, and four others dropped out because of side effects. During the follow up period (mean 10.9 (SD 5.8) months), 29 "responders" needed a mean application of capsaicin every day (1.6 (SD 1.2); range 0.5-7 days) to remain symptom free (or nearly symptom free). CONCLUSION: Capsaicin is a new, safe, and highly effective treatment for severe intractable idiopathic pruritus ani. PMID- 12912866 TI - Upregulation of lymphotoxin beta expression in liver progenitor (oval) cells in chronic hepatitis C. AB - BACKGROUND: Bipotent liver progenitor (oval) cells with the ability to differentiate into hepatocytes and biliary epithelium have recently been identified in human subjects with hepatitis C. Animal studies suggest that members of the tumour necrosis factor family, including lymphotoxin beta (LT beta), regulate oval cell proliferation in liver disease, but its role in human liver disease is unclear. AIMS: This study seeks to establish a role for LT-beta in hepatitis C related liver injury and to provide evidence that its increased expression is related to the presence of oval cells. METHODS: Liver biopsy specimens were obtained from patients with chronic hepatitis C virus (HCV) infection (n=20). Control liver samples (n=5) were obtained from liver resection or transplant surgery. LT-beta expression in liver biopsy specimens was studied using quantitative real time polymerase chain reaction and immunohistochemistry. RESULTS: LT-beta mRNA levels were similar in control and HCV liver in the absence of fibrosis. In subjects with portal fibrosis, LT-beta mRNA levels were elevated 2.2-fold over control liver levels (p=0.04). In subjects with bridging fibrosis, LT-beta mRNA levels increased 4.4-fold over control liver levels (p=0.02). LT beta mRNA levels in subjects with established cirrhosis were increased 3.3-fold compared with controls and 2.6-fold compared with mild liver damage (p=0.02). Immunohistochemical analysis established that LT-beta was expressed by oval cells, inflammatory cells, and small portal hepatocytes. CONCLUSIONS: In chronic HCV infection, LT-beta expression is observed in multiple hepatic cell types, including oval cells. LT-beta expression is significantly increased when fibrosis or cirrhosis is present, suggesting a role for LT-beta in the pathogenesis of chronic hepatitis C and a possible role in oval cell mediated liver regeneration. PMID- 12912867 TI - Risk of hepatitis C virus transmission from patients to surgeons: model based on an unlinked anonymous study of hepatitis C virus prevalence in hospital patients in Glasgow. AB - BACKGROUND: The risk of a surgeon acquiring the hepatitis C virus (HCV) through occupational exposure is dependent on the prevalence of HCV infection in the patient population, the probability of a percutaneous injury transmitting HCV, and the incidence of percutaneous injury during surgery. AIMS: To estimate the prevalence of HCV infection in the adult surgical patient population in North Glasgow and thereafter estimate the risk of HCV transmission to surgeons through occupational exposure. METHODS: The prevalence of HCV infection was estimated through the unlinked anonymous testing of samples from male surgical patients, aged 16-49 years, in two North Glasgow hospitals from 1996 to 1997, and adjusting these data for age and sex. Using published estimates of the incidence of percutaneous injury during surgery and percutaneous injury transmitting HCV, the risk of occupational transmission of HCV to surgeons was then derived. RESULTS: The estimated prevalence of anti-HCV infection for all adult patients in the two hospitals combined was 1.4% (cardiothoracic/cardiology 0.8%, orthopaedics/rheumatology 1.4%, general surgery/ENT 2.0%). The estimated probability of HCV transmission from an HCV infected patient to an uninfected surgeon was 0.001-0.032% per annum (0.035-1.12% risk over a 35 year professional career). CONCLUSIONS: The risk of an individual surgeon acquiring HCV through occupational exposure is low, even in an area with an extremely high prevalence of HCV among its injecting drug using population. Surgeons however should be encouraged to observe universal precautions and present for assessment after needlestick injuries to protect themselves and their patients from this insidious infection. PMID- 12912868 TI - Location and function of intrahepatic shunts in anaesthetised rats. AB - BACKGROUND: In the present study we determined the proportion of shunt flow due to patent intrahepatic portal systemic shunts in the normal rat liver and its relationship with microsphere induced portal hypertension. METHODS: Systemic and hepatic haemodynamics were measured continuously before, during, and after intraportal injection of 15 micro m diameter microspheres in anaesthetised male Wistar rats. Functional hepatic blood flow and intrahepatic shunt flow were determined by the use of constant intraportal infusion of sorbitol and simultaneous measurements in the portal vein, hepatic vein, and carotid artery. The percentage of large shunts of diameter >15 micro m were estimated by intraportal injection of (51)Cr labelled 15 micro m diameter microspheres. RESULTS: Hepatic sorbitol uptake was 97.9 (0.5)% in normal control rats, with functional hepatic blood flow equalling total hepatic blood flow (2.52 (0.23) ml/min/100 g body weight). Microsphere injection decreased sorbitol uptake to 12.8 (4.3)% and further to 4.1 (0.7)% when followed by hepatic arterial ligation. In the latter two groups, intrahepatic shunt flow (1.46 (0.15) and 1.16 (0.19) ml/min/100 g body weight, respectively) was not significantly different from portal venous flow (1.36 (0.20) and 1.20 (0.20) ml/min/100 g body weight, respectively). Portal venous flow remained at 70% of basal values and portal venous pressure only increased by 50% from baseline. (51)Cr labelled microsphere shunt fraction through large shunts (>15 micro m) was less than 1.0%. CONCLUSION: The site of confluence between the hepatic artery and portal vein is in zone II. Intrahepatic shunts originate in presinusoidal regions in zone I in the normal liver and, when activated by intraportal injection of microspheres, divert 70% of the total portal blood flow away from zone III and thereby reduce acute increases in portal venous pressure. PMID- 12912869 TI - Vascular endothelial growth factor and receptor interaction is a prerequisite for murine hepatic fibrogenesis. AB - BACKGROUND: It has been shown that expression of the potent angiogenic factor, vascular endothelial growth factor (VEGF), and its receptors, flt-1 (VEGFR-1) and KDR/Flk-1 (VEGFR-2), increased during the development of liver fibrosis. AIMS: To elucidate the in vivo role of interaction between VEGF and its receptors in liver fibrogenesis. METHODS: A model of CCl(4) induced hepatic fibrosis was used to assess the role of VEGFR-1 and VEGFR-2 by means of specific neutralising monoclonal antibodies (R-1mAb and R-2mAb, respectively). R-1mAb and R-2mAb were administered after two weeks of treatment with CCl(4), and indices of fibrosis were assessed at eight weeks. RESULTS: Hepatic VEGF mRNA expression significantly increased during the development of liver fibrosis. Both R-1mAb and R-2mAb treatments significantly attenuated the development of fibrosis associated with suppression of neovascularisation in the liver. Hepatic hydroxyproline and serum fibrosis markers were also suppressed. Furthermore, the number of alpha-smooth muscle actin positive cells and alpha1(I)-procollagen mRNA expression were significantly suppressed by R-1mAb and R-2mAb treatment. The inhibitory effect of R-2mAb was more potent than that of R-1mAb, and combination treatment with both mAbs almost completely attenuated fibrosis development. Our in vitro study showed that VEGF treatment significantly stimulated proliferation of both activated hepatic stellate cells (HSC) and sinusoidal endothelial cells (SEC). VEGF also significantly increased alpha1(I)-procollagen mRNA expression in activated HSC. CONCLUSIONS: These results suggest that the interaction of VEGF and its receptor, which reflected the combined effects of both on HSC and SEC, was a prerequisite for liver fibrosis development. PMID- 12912870 TI - Portopulmonary hypertension in decompensated cirrhosis with refractory ascites. AB - BACKGROUND: The prevalence of portopulmonary hypertension (PPHTN) in patients with cirrhosis and refractory ascites is unknown. Its presence may preclude patients from receiving a transjugular intrahepatic portosystemic shunt or liver transplantation as a definitive treatment for their end stage cirrhosis. PURPOSE: To determine the prevalence, possible aetiological factors, and predictive factors for the development of PPHTN in these patients. METHODS: Sixty two patients (53 males, nine females; mean age 54.5 (1.4) years) with biopsy proven cirrhosis and refractory ascites underwent angiographic measurements of pulmonary and splanchnic haemodynamics. Endothelin 1 levels were measured from the pulmonary artery. Forty nine patients underwent radionuclide angiography for measurements of central blood volume, pulmonary vascular, and cardiac chamber volumes. Forty seven patients also underwent two dimensional echocardiography for measurements of cardiac structural and functional parameters. Cardiac output, and systemic and pulmonary vascular resistance were calculated. RESULTS: Ten patients (16.1%) fulfilled the criteria for PPHTN (mean pulmonary artery pressure >/= 25 mm Hg and pulmonary vascular resistance >/= 120 dynxs/cm(5)), with significantly higher mean right atrial (15.4 (1.2) v 7.9 (0.5) mm Hg; p<0.001), and right ventricular pressures (24.7 (1.5) v 14.7 (0.6) mm Hg; p<0.001), and endothelin 1 levels (3.04 (0.40) v 1.98 (0.12) pg/ml; p=0.02). No significant differences in any of the other parameters measured were detected between the two groups. A right atrial pressure of >/= 14 mm Hg had a 83% positive predictive value for the presence of PPHTN. CONCLUSIONS: Portopulmonary hypertension is common in cirrhosis with refractory ascites, possibly due to excess endothelin 1 in the pulmonary circulation. An elevated right atrial pressure >/= 14 mm Hg predicts the presence of PPHTN, which may be helpful in deciding management options in these patients. PMID- 12912871 TI - Glucocorticoid receptors are downregulated in hepatic T lymphocytes in rats with experimental cholangitis. AB - BACKGROUND: and aims: Primary sclerosing cholangitis is a Th1 cytokine driven disease with a poor clinical responsiveness to glucocorticoid therapy. We have previously documented elevated circulating glucocorticoid levels in cholestatic rats and in addition have noted increased hepatic expression of the Th1 cytokine interferon gamma (IFN-gamma) in a rat model of cholangitis. Therefore, we examined the relationship between circulating glucocorticoid levels, hepatic IFN gamma expression, and hepatic T cell glucocorticoid receptor (GR) expression in a rat model of cholangitis to provide insight into the possible mechanism underlying hepatic T cell glucocorticoid resistance in cholangitic diseases. METHODS: Cholangitis was induced in male Sprague-Dawley rats by oral administration of low dose alpha-naphthylisothiocyanate (ANIT). On day 14, ANIT fed and control rats were sacrificed, serum collected, and hepatic, splenic, and peripheral blood T lymphocytes isolated for GR expression, as determined by reverse transcription-polymerase chain reaction and western blotting. RESULTS: Circulating glucocorticoid levels were markedly elevated in ANIT fed rats. Hepatic T lymphocyte GR mRNA and protein levels were significantly reduced in ANIT treated rats compared with controls. In contrast, GR mRNA and protein expression in splenic and circulating T lymphocytes was similar in both groups. Furthermore, reduced hepatic T cell GR expression in ANIT fed rats was associated with reduced hepatic CD4(+) T cell sensitivity to dexamethasone inhibitory effects (that is, inhibition of interleukin 2 receptor expression). CONCLUSION: We conclude that hepatic T lymphocyte resistance to elevated endogenous glucocorticoid levels in rats with experimental cholangitis appears, in part, to be mediated by decreased GR expression. PMID- 12912872 TI - Intestinal absorption of the bile acid analogue 75Se-homocholic acid-taurine is increased in primary biliary cirrhosis, and reverts to normal during ursodeoxycholic acid administration. AB - BACKGROUND: Whether ileal absorption of bile acid is up or downregulated in chronic cholestasis is still debated, and most evidence has come from animal studies. AIMS: To compare ileal bile acid absorption in patients with primary biliary cirrhosis (PBC) and in healthy control subjects, and to assess the effect of ursodeoxycholic acid (UDCA). PATIENTS: We studied 14 PBC patients before and during (n=11) UDCA administration, 14 healthy control subjects, and 14 Crohn's disease patients (as disease controls). METHODS: We used cholescintigraphy to measure retention in the enterohepatic circulation over five successive days of the bile acid analogue (75)Se-homocholic acid-taurine ((75)SeHCAT) as an index of ileal bile acid absorption. Results were expressed as (75)SeHCAT fractional turnover rate (FTR) and t(1/2)12. RESULTS: (75)SeHCAT FTR was 0.19 (0.11)/day, 0.34 (0.11)/day (p<0.001), and 0.83 (0.32)/day in PBC patients, healthy controls (p<0.0001), and Crohn's patients (p<0.001), respectively, which increased to 0.36 (0.16)/day in PBC patients during UDCA treatment (p<0.005). (75)SeHCAT t(1/2)12 was 4.8 (2.1) days in PBC patients, 2.2 (0.5) days (p<0.001) in healthy controls, and 1.0 (0.5) days (p<0.001) in Crohn's disease patients. (75)SeHCAT t(1/2)12 decreased to 2.2 (0.93) days (p< 0.001) in PBC patients during UDCA treatment. CONCLUSIONS: Our results support the concept that ileal bile acid absorption is upregulated in PBC patients, and that this effect may contribute towards damaging the cholestatic liver. This upregulation of bile acid absorption is abolished by UDCA. PMID- 12912873 TI - NSAIDs: the emperor's new dogma? AB - The spectacular marketing success of the selective cyclooxygenase 2 (COX-2) inhibitors is largely based on efficacy comparable with conventional non steroidal anti-inflammatory drugs (NSAIDs) with vastly improved gastrointestinal safety. The additional key to the marketing success is the purity and simplicity of the message-that is, COX-1 inhibition causes the gastrointestinal side effects of NSAIDs (COX-1 dogma) while COX-2 blocking confers the therapeutic benefits (COX-2 dogma). Adherence to the COX dogmas with development of COX-2 selective agents has undoubtedly benefited many patients, but ironically their scientific basis is now seriously challenged by experimentation. PMID- 12912874 TI - New dogmas or old? AB - Recent experimental studies may undermine our understanding of the gastrointestinal side effects of non-steroidal anti-inflammatory drugs and cast a shadow on the original concept that underpins the development of the recent addition to the clinical anti-inflammatory armamentarium, the COX-2 selective inhibitors. But is this just a passing cloud or a total eclipse of the COX theory? PMID- 12912876 TI - Meeting report of the British Society for Gastroenterology meeting, March 2003. PMID- 12912877 TI - Is hepatobiliary scintigraphy indeed insensitive for the diagnosis of sphincter of Oddi dysfunction? PMID- 12912878 TI - Is hepatobiliary scintigraphy insensitive for the diagnosis of sphincter of Oddi dysfunction? PMID- 12912879 TI - Long term follow up of Helicobacter pylori induced gastric diffuse large B cell MALT lymphoma following eradication treatment alone. PMID- 12912880 TI - Worsening of steatosis and fibrosis progression. PMID- 12912881 TI - Atrophic gastritis: pathology and endoscopy in the reversibility assessment. PMID- 12912882 TI - Colorectal screening guidelines in acromegaly. PMID- 12912883 TI - Diversion colitis with a mucosal tear on endoscopic insufflation. PMID- 12912885 TI - Genome-wide expression analyses of gene regulation during early development of Dictyostelium discoideum. AB - Using genome-wide microarrays, we recognized 172 genes that are highly expressed at one stage or another during multicellular development of Dictyostelium discoideum. When developed in shaken suspension, 125 of these genes were expressed if the cells were treated with cyclic AMP (cAMP) pulses at 6-min intervals between 2 and 6 h of development followed by high levels of exogenous cAMP. In the absence of cAMP treatment, only three genes, carA, gbaB, and pdsA, were consistently expressed. Surprisingly, 14 other genes were induced by cAMP treatment of mutant cells lacking the activatable adenylyl cyclase, ACA. However, these genes were not cAMP induced if both of the developmental adenylyl cyclases, ACA and ACR, were disrupted, showing that they depend on an internal source of cAMP. Constitutive activity of the cAMP-dependent protein kinase PKA was found to bypass the requirement of these genes for adenylyl cyclase and cAMP pulses, demonstrating the critical role of PKA in transducing the cAMP signal to early gene expression. In the absence of constitutive PKA activity, expression of later genes was strictly dependent on ACA in pulsed cells. PMID- 12912884 TI - A yeast under cover: the capsule of Cryptococcus neoformans. PMID- 12912886 TI - Sequence elements in both the intergenic space and the 3' untranslated region of the Crithidia fasciculata KAP3 gene are required for cell cycle regulation of KAP3 mRNA. AB - mRNA levels of several Crithidia fasciculata genes involved in DNA metabolism have previously been found to cycle as cells progress through the cell cycle. Octamer consensus sequences in the 5' untranslated regions (5' UTRs) of these transcripts were shown to be required for cycling of these mRNAs. The KAP3 gene encodes a kinetoplast histone H1-like DNA binding protein, and its mRNA levels cycle in parallel with those of the kinetoplast DNA topoisomerase (TOP2), dihydrofolate reductase-thymidylate synthase (DHFR-TS), and the large subunit of the nuclear single-stranded DNA binding protein (RPA1). KAP3 mRNA contains two octamer consensus sequences in its 3' UTR but none in its 5' UTR. Mutation of these octamer sequences was not sufficient to prevent cycling of a sequence tagged KAP3 mRNA expressed from a plasmid. Mutation of an octamer sequence contained on the precursor transcript but not on the mRNA, in addition to mutation of the two octamer sequences in the 3' UTR, was necessary to abolish cycling of the mRNA. The requirement for a sequence not present on the mature mRNA indicates that regulation of the mRNA levels by the octamer sequences occurs at or prior to splicing of the transcript. Incompletely processed RNAs containing octamer sequences were also found to accumulate during the cell cycle when the mRNA levels were lowest. These RNA species hybridize to both the KAP3 coding sequence and that of the downstream drug resistance gene, indicating a lack of processing within the intergenic region separating these genes. We propose a cell cycle-dependent interference in transcript processing mediated by octamer consensus sequences as a mechanism contributing to the cycling of such transcripts. PMID- 12912887 TI - Diverse sequences within Tlr elements target programmed DNA elimination in Tetrahymena thermophila. AB - Tlr elements are a novel family of approximately 30 putative mobile genetic elements that are confined to the germ line micronuclear genome in Tetrahymena thermophila. Thousands of diverse germ line-limited sequences, including the Tlr elements, are specifically eliminated from the differentiating somatic macronucleus. Macronucleus-retained sequences flanking deleted regions are known to contain cis-acting signals that delineate elimination boundaries. It is unclear whether sequences within deleted DNA also play a regulatory role in the elimination process. In the current study, an in vivo DNA rearrangement assay was used to identify internal sequences required in cis for the elimination of Tlr elements. Multiple, nonoverlapping regions from the approximately 23-kb Tlr elements were independently sufficient to stimulate developmentally regulated DNA elimination when placed within the context of flanking sequences from the most thoroughly characterized family member, Tlr1. Replacement of element DNA with macronuclear or foreign DNA abolished elimination activity. Thus, diverse sequences dispersed throughout Tlr DNA contain cis-acting signals that target these elements for programmed elimination. Surprisingly, Tlr DNA was also efficiently deleted when Tlr1 flanking sequences were replaced with DNA from a region of the genome that is not normally associated with rearrangement, suggesting that specific flanking sequences are not required for the elimination of Tlr element DNA. PMID- 12912888 TI - Mannitol is required for stress tolerance in Aspergillus niger conidiospores. AB - D-Mannitol is the predominant carbon compound in conidiospores of the filamentous fungus Aspergillus niger and makes up 10 to 15% of the dry weight. A number of physiological functions have been ascribed to mannitol, including serving as a reserve carbon source, as an antioxidant, and to store reducing power. In this study, we cloned and characterized the A. niger mpdA gene, which encodes mannitol 1-phosphate dehydrogenase (MPD), the first enzyme in the mannitol biosynthesis pathway. The mpdA promoter contains putative binding sites for the development specific transcription factors BRLA and ABAA. Furthermore, increased expression of mpdA in sporulating mycelium suggests that mannitol biosynthesis is, to a certain extent, developmentally regulated in A. niger. Inactivation of mpdA abolished mannitol biosynthesis in growing mycelium and reduced the mannitol level in conidiospores to 30% that in the wild type, indicating that MPD and mannitol 1-phosphate phosphatase form the major metabolic pathway for mannitol biosynthesis in A. niger. The viability of spores after prolonged storage and germination kinetics were normal in an mpdA null mutant, indicating that mannitol does not play an essential role as a reserve carbon source in A. niger conidia. However, conidiospores of a DeltampdA strain were extremely sensitive to a variety of stress conditions, including high temperature, oxidative stress and, to a lesser extent, freezing and lyophilization. Since mannitol supplied in the medium during sporulation repaired this deficiency, mannitol appears to be essential for the protection of A. niger spores against cell damage under these stress conditions. PMID- 12912889 TI - Environmental suppression of Neurospora crassa cot-1 hyperbranching: a link between COT1 kinase and stress sensing. AB - cot-1 mutants belong to a class of Neurospora crassa colonial temperature sensitive (cot) mutants that exhibit abnormal polar extension and branching patterns when grown at restrictive temperatures. cot-1 encodes a Ser/Thr protein kinase that is structurally related to the human myotonic dystrophy kinase which, when impaired, confers a disease that involves changes in cytoarchitecture and ion homeostasis. When grown under restrictive conditions, cot-1 cultures exhibited enhanced medium acidification rates, increased relative abundance of sodium, and increased intracellular glycerol content, indicating an ion homeostasis defect in a hyperbranching mutant. The application of ion transport blockers led to only mild suppression of the cot-1 phenotype. The presence of increased medium NaCl or sorbitol, H(2)O(2), or ethanol levels significantly suppressed the cot-1 phenotype, restored ion homeostasis, and was accompanied by reduced levels of cyclic AMP-dependent protein kinase (PKA) activity. The cot-1 phenotype could also be partially suppressed by direct inhibition of PKA with KT 5720. A reduced availability of fermentable carbon sources also had a suppressive effect on the cot-1 phenotype. In contrast to the effect of extragenic ropy suppressors of cot-1, environmental stress-related suppression of cot-1 did not change COT1 polypeptide expression patterns in the mutant. We suggest that COT1 function is linked to environmental stress response signaling and that altering PKA activity bypasses the requirement for fully functional COT1. PMID- 12912890 TI - Glucose uptake in Trichoderma harzianum: role of gtt1. AB - Using a differential display technique, the gene gtt1, which codes for a high affinity glucose transporter, has been cloned from the mycoparasite fungus Trichoderma harzianum CECT 2413. The deduced protein sequence of the gtt1 gene shows the 12 transmembrane domains typical of sugar transporters, together with certain residues involved in glucose uptake, such as a conserved arginine between domains IV and V and an aromatic residue (Phe) in the sequence of domain X. The gtt1 gene is transcriptionally regulated, being repressed at high levels of glucose. When carbon sources other than glucose are utilized, gtt1 repression is partially alleviated. Full derepression of gtt1 is obtained when the fungus is grown in the presence of low carbon source concentrations. This regulation pattern correlates with the role of this gene in glucose uptake during carbon starvation. Gene expression is also controlled by pH, so that the gtt1 gene is repressed at pH 6 but not at pH 3, a fact which represents a novel aspect of the influence of pH on the gene expression of transporters. pH also affects glucose transport, since a strongly acidic pH provokes a 40% decrease in glucose transport velocity. Biochemical characterization of the transport shows a very low K(m) value for glucose (12 micro M). A transformant strain that overexpresses the gtt1 gene shows a threefold increase in glucose but not galactose or xylose uptake, a finding which confirms the role of the gtt1 gene in glucose transport. The cloning of the first filamentous ascomycete glucose transporter is the first step in elucidating the mechanisms of glucose uptake and carbon repression in aerobic fungi. PMID- 12912891 TI - Diverged binding specificity of Rim101p, the Candida albicans ortholog of PacC. AB - The biology of Candida albicans, including dimorphism and virulence, is significantly influenced by environmental pH. The response to ambient pH includes the pH-conditional expression of several genes, which is directly or indirectly regulated by Rim101p. Rim101p is homologous to PacC, a transcription factor that regulates pH-conditional gene expression in Aspergillus nidulans. PacC binds 5' GCCARG-3' sequences upstream of pH-responsive genes and either activates or represses transcription. The absence of pacC consensus binding sites upstream of PHR1, a RIM101-dependent, alkaline pH-induced gene of C. albicans, suggested either that PHR1 is indirectly regulated by Rim101p or that the binding specificity of Rim101p is different. In vitro binding studies demonstrated that Rim101p strongly bound two regions upstream of PHR1 that were only weakly bound by PacC. Deletion analysis and site-specific mutagenesis demonstrated that both sites were functionally significant, mutation of either site reduced RIM101 dependent induction, and expression was abolished in the double mutant. Furthermore, oligonucleotides containing these sites conferred pH-conditional expression when inserted upstream of a reporter gene. The consensus sequence of these sites, 5'-CCAAGAAA-3', was identical to the binding recognition sequence identified by in vitro selection of Rim101p binding oligonucleotides from a random pool. The functional significance of this binding sequence was reinforced by its observed presence upstream of a number of newly identified pH-conditional genes. We conclude that Rim101p acts as a transcription factor and directly regulates pH-conditional gene expression but has a binding specificity different from that of PacC. PMID- 12912892 TI - Inositol and phosphate regulate GIT1 transcription and glycerophosphoinositol incorporation in Saccharomyces cerevisiae. AB - Glycerophosphoinositol is produced through deacylation of the essential phospholipid phosphatidylinositol. In Saccharomyces cerevisiae, the glycerophosphoinositol produced is excreted from the cell but is recycled for phosphatidylinositol synthesis when inositol is limiting. To be recycled, glycerophosphoinositol enters the cell through the permease encoded by GIT1. The transport of exogenous glycerophosphoinositol through Git1p is sufficiently robust to support the growth of an inositol auxotroph (ino1Delta). We now report that S. cerevisiae also uses exogenous phosphatidylinositol as an inositol source. Evidence suggests that phosphatidylinositol is deacylated to glycerophosphoinositol extracellularly before being transported across the plasma membrane by Git1p. A genetic screen identified Pho86p, which is required for targeting of the major phosphate transporter (Pho84p) to the plasma membrane, as affecting the utilization of phosphatidylinositol and glycerophosphoinositol. Deletion of PHO86 in an ino1Delta strain resulted in faster growth when either phosphatidylinositol or glycerophosphoinositol was supplied as the sole inositol source. The incorporation of radiolabeled glycerophosphoinositol into an ino1Delta pho86Delta mutant was higher than that into wild-type, ino1Delta, and pho86Delta strains. All strains accumulated the most GIT1 transcript when incubated in media limited for inositol and phosphate in combination. However, the ino1Delta pho86Delta mutant accumulated approximately threefold more GIT1 transcript than did the other strains when incubated in inositol-free media containing either high or low concentrations of P(i). Deletion of PHO4 abolished GIT1 transcription in a wild-type strain. These results indicate that the transport of glycerophosphoinositol by Git1p is regulated by factors affecting both inositol and phosphate availabilities and suggest a regulatory connection between phosphate metabolism and phospholipid metabolism. PMID- 12912893 TI - PAK paradox: Paramecium appears to have more K(+)-channel genes than humans. AB - K(+)-selective ion channels (K(+) channels) have been found in bacteria, archaea, eucarya, and viruses. In Paramecium and other ciliates, K(+) currents play an essential role in cilia-based motility. We have retrieved and sequenced seven closely related Paramecium K(+)-channel gene (PAK) sequences by using previously reported fragments. An additional eight unique K(+)-channel sequences were retrieved from an indexed library recently used in a pilot genome sequencing project. Alignments of these protein translations indicate that while these 15 genes have diverged at different times, they all maintain many characteristics associated with just one subclass of metazoan K(+) channels (CNG/ERG type). Our results indicate that most of the genes are expressed, because all predicted frameshifts and several gaps in the homolog alignments contain Paramecium intron sequences deleted from reverse transcription-PCR products. Some of the variations in the 15 genomic nucleotide sequences involve an absence of introns, even between very closely related sequences, suggesting a potential occurrence of reverse transcription in the past. Extrapolation from the available genome sequence indicates that Paramecium harbors as many as several hundred of this one type of K(+)-channel gene. This quantity is far more numerous than those of K(+) channel genes of all types known in any metazoan (e.g., approximately 80 in humans, approximately 30 in flies, and approximately 15 in Arabidopsis). In an effort to understand this plurality, we discuss several possible reasons for their maintenance, including variations in expression levels in response to changes in the freshwater environment, like that seen with other major plasma membrane proteins in Paramecium. PMID- 12912894 TI - Roles of Candida albicans Dfg5p and Dcw1p cell surface proteins in growth and hypha formation. AB - The Candida albicans cell wall participates in both growth and morphological transitions between yeast and hyphae. Our studies here focus on Dfg5p and Dcw1p, two similar proteins with features of glycosylphosphatidylinositol-linked cell surface proteins. Mutants lacking Dfg5p are defective in alkaline pH-induced hypha formation; mutants lacking Dcw1p have no detected hypha formation defect. Both homozygote-triplication tests and conditional expression strategies indicate that dfg5 and dcw1 mutations are synthetically lethal. Therefore, Dfg5p and Dcw1p share a function required for growth. Epitope-tagged Dfg5p, created through an insertional mutagenesis strategy, is found in cell membrane and cell wall extract fractions, and endoglycosidase H digestion shows that Dfg5p undergoes N-linked mannosylation. Surprisingly, Dfg5p is required for expression of the hypha specific gene HWP1 in alkaline media. Because Dfg5p is a cell surface protein, it is poised to generate or transmit an external signal required for the program of hypha-specific gene expression. PMID- 12912895 TI - Formation and remodeling of inositolphosphoceramide during differentiation of Trypanosoma cruzi from trypomastigote to amastigote. AB - Differentiation of Trypanosoma cruzi trypomastigotes to amastigotes inside myoblasts or in vitro, at low extracellular pH, in the presence of [(3)H]palmitic acid or [(3)H]inositol revealed differential labeling of inositolphosphoceramide and phosphatidylinositol, suggesting that a remodeling process takes place in both lipids. Using (3)H-labeled inositolphosphoceramide and phosphatidylinositol as substrates, we demonstrated the association of at least five enzymatic activities with the membranes of amastigotes and trypomastigotes. These included phospholipase A(1), phospholipase A(2), inositolphosphoceramide-fatty acid hydrolase, acyltransferase, and a phospholipase C releasing either ceramide or a glycerolipid from the inositolphospholipids. These enzymes may be acting in remodeling reactions leading to the anchor of mature glycoproteins or glycoinositolphospholipids and helping in the transformation of the plasma membrane, a necessary step in the differentiation of slender trypomastigotes to round amastigotes. Synthesis of inositolphosphoceramide and particularly of glycoinositolphospholipids was inhibited by aureobasidin A, a known inhibitor of fungal inositolphosphoceramide synthases. The antibiotic impaired the differentiation of trypomastigotes at acidic pH, as indicated by an increased appearance of intermediate forms and a decreased expression of the Ssp4 glycoprotein, a characteristic marker of amastigote forms. Aureobasidin A was also toxic to differentiating trypomastigotes at acidic pH but not to trypomastigotes maintained at neutral pH. Our data suggest that inositolphosphoceramide is implicated in T. cruzi differentiation and that its metabolism could provide important targets for the development of antiparasitic therapies. PMID- 12912896 TI - Protein kinase involved in flagellar-length control. AB - During its life cycle, the parasitic protozoon Leishmania mexicana differentiates from a flagellated form, the promastigote, to an amastigote form carrying a rudimentary flagellum. Besides biochemical changes, this process involves a change in overall cell morphology including flagellar shortening. A mitogen activated protein kinase kinase homologue designated LmxMKK was identified in a homology screening and found to be critically involved in the regulation of flagellar assembly and cell size. LmxMKK is exclusively expressed in the promastigote stage and is likely to be regulated by posttranslational mechanisms such as phosphorylation. A deletion mutant for the single-copy gene revealed motile flagella dramatically reduced in length and lacking the paraflagellar rod, a structure adjacent to the axoneme in kinetoplastid flagella. Moreover, a fraction of the cells showed perturbance of the axonemal structure. Complementation of the deletion mutant with the wild-type gene restored typical promastigote morphology. We propose that LmxMKK influences anterograde intraflagellar transport to maintain flagellar length in Leishmania promastigotes; as such, it is the first protein kinase known to be involved in organellar assembly. PMID- 12912897 TI - Formation of Hirano bodies induced by expression of an actin cross-linking protein with a gain-of-function mutation. AB - Hirano bodies are paracrystalline actin filament-containing structures reported to be associated with a variety of neurodegenerative diseases. However, the biological function of Hirano bodies remains poorly understood, since nearly all prior studies of these structures were done with postmortem samples of tissue. In the present study, we generated a full-length form of a Dictyostelium 34-kDa actin cross-linking protein with point mutations in the first putative EF hand, termed 34-kDa DeltaEF1. The 34-kDa DeltaEF1 protein binds calcium normally but has activated actin binding that is unregulated by calcium. The expression of the 34-kDa DeltaEF1 protein in Dictyostelium induces the formation of Hirano bodies, as assessed by both fluorescence microscopy and transmission electron microscopy. Dictyostelium cells bearing Hirano bodies grow normally, indicating that Hirano bodies are not associated with cell death and are not deleterious to cell growth. Moreover, the expression of the 34-kDa DeltaEF1 protein rescues the phenotypes of cells lacking the 34-kDa protein and cells lacking both the 34-kDa protein and alpha-actinin. Finally, the expression of the 34-kDa DeltaEF1 protein also initiates the formation of Hirano bodies in cultured mouse fibroblasts. These results show that the failure to regulate the activity and/or affinity of an actin cross-linking protein can provide a signal for the formation of Hirano bodies. More generally, the formation of Hirano bodies is a cellular response to or a consequence of aberrant function of the actin cytoskeleton. PMID- 12912898 TI - CF45-1, a secreted protein which participates in Dictyostelium group size regulation. AB - Developing Dictyostelium cells aggregate to form fruiting bodies containing typically 2 x 10(4) cells. To prevent the formation of an excessively large fruiting body, streams of aggregating cells break up into groups if there are too many cells. The breakup is regulated by a secreted complex of polypeptides called counting factor (CF). Countin and CF50 are two of the components of CF. Disrupting the expression of either of these proteins results in cells secreting very little detectable CF activity, and as a result, aggregation streams remain intact and form large fruiting bodies, which invariably collapse. We find that disrupting the gene encoding a third protein present in crude CF, CF45-1, also results in the formation of large groups when cells are grown with bacteria on agar plates and then starve. However, unlike countin(-) and cf50(-) cells, cf45 1(-) cells sometimes form smaller groups than wild-type cells when the cells are starved on filter pads. The predicted amino acid sequence of CF45-1 has some similarity to that of lysozyme, but recombinant CF45-1 has no detectable lysozyme activity. In the exudates from starved cells, CF45-1 is present in a approximately 450-kDa fraction that also contains countin and CF50, suggesting that it is part of a complex. Recombinant CF45-1 decreases group size in colonies of cf45-1(-) cells with a 50% effective concentration (EC(50)) of approximately 8 ng/ml and in colonies of wild-type and cf50(-) cells with an EC(50) of approximately 40 ng/ml. Like countin(-) and cf50(-) cells, cf45-1(-) cells have high levels of cytosolic glucose, high cell-cell adhesion, and low cell motility. Together, the data suggest that CF45-1 participates in group size regulation in Dictyostelium. PMID- 12912899 TI - Asexual development is increased in Neurospora crassa cat-3-null mutant strains. AB - We use asexual development of Neurospora crassa as a model system with which to determine the causes of cell differentiation. Air exposure of a mycelial mat induces hyphal adhesion, and adherent hyphae grow aerial hyphae that, in turn, form conidia. Previous work indicated the development of a hyperoxidant state at the start of these morphogenetic transitions and a large increase in catalase activity during conidiation. Catalase 3 (CAT-3) increases at the end of exponential growth and is induced by different stress conditions. Here we analyzed the effects of cat-3-null strains on growth and asexual development. The lack of CAT-3 was not compensated by other catalases, even under oxidative stress conditions, and cat-3(RIP) colonies were sensitive to H(2)O(2), indicating that wild-type (Wt) resistance to external H(2)O(2) was due to CAT-3. cat-3(RIP) colonies grown in the dark produced high levels of carotenes as a consequence of oxidative stress. Light exacerbated oxidative stress and further increased carotene synthesis. In the cat-3(RIP) mutant strain, increased aeration in liquid cultures led to increased hyphal adhesion and protein oxidation. Compared to the Wt, the cat-3(RIP) mutant strain produced six times more aerial hyphae and conidia in air-exposed mycelial mats, as a result of longer and more densely packed aerial hyphae. Protein oxidation in colonies was threefold higher and showed more aerial hyphae and conidia in mutant strains than did the Wt. Results indicate that oxidative stress due to lack of CAT-3 induces carotene synthesis, hyphal adhesion, and more aerial hyphae and conidia. PMID- 12912901 TI - Specific protein targeting during cell differentiation: polarized localization of Fus1p during mating depends on Chs5p in Saccharomyces cerevisiae. AB - In budding yeast, chs5 mutants are defective in chitin synthesis and cell fusion during mating. Chs5p is a late-Golgi protein required for the polarized transport of the chitin synthase Chs3p to the membrane. Here we show that Chs5p is also essential for the polarized targeting of Fus1p, but not of other cell fusion proteins, to the membrane during mating. PMID- 12912900 TI - POS5 gene of Saccharomyces cerevisiae encodes a mitochondrial NADH kinase required for stability of mitochondrial DNA. AB - In a search for nuclear genes that affect mutagenesis of mitochondrial DNA in Saccharomyces cerevisiae, an ATP-NAD (NADH) kinase, encoded by POS5, that functions exclusively in mitochondria was identified. The POS5 gene product was overproduced in Escherichia coli and purified without a mitochondrial targeting sequence. A direct biochemical assay demonstrated that the POS5 gene product utilizes ATP to phosphorylate both NADH and NAD(+), with a twofold preference for NADH. Disruption of POS5 increased minus-one frameshift mutations in mitochondrial DNA 50-fold, as measured by the arg8(m) reversion assay, with no increase in nuclear mutations. Also, a dramatic increase in petite colony formation and slow growth on glycerol or limited glucose were observed. POS5 was previously described as a gene required for resistance to hydrogen peroxide. Consistent with a role in the mitochondrial response to oxidative stress, a pos5 deletion exhibited a 28-fold increase in oxidative damage to mitochondrial proteins and hypersensitivity to exogenous copper. Furthermore, disruption of POS5 induced mitochondrial biogenesis as a response to mitochondrial dysfunction. Thus, the POS5 NADH kinase is required for mitochondrial DNA stability with a critical role in detoxification of reactive oxygen species. These results predict a role for NADH kinase in human mitochondrial diseases. PMID- 12912902 TI - Crystal structure and functional analysis of Escherichia coli glutamate decarboxylase. AB - Glutamate decarboxylase is a vitamin B6-dependent enzyme, which catalyses the decarboxylation of glutamate to gamma-aminobutyrate. In Escherichia coli, expression of glutamate decarboxylase (GadB), a 330 kDa hexamer, is induced to maintain the physiological pH under acidic conditions, like those of the passage through the stomach en route to the intestine. GadB, together with the antiporter GadC, constitutes the gad acid resistance system, which confers the ability for bacterial survival for at least 2 h in a strongly acidic environment. GadB undergoes a pH-dependent conformational change and exhibits an activity optimum at low pH. We determined the crystal structures of GadB at acidic and neutral pH. They reveal the molecular details of the conformational change and the structural basis for the acidic pH optimum. We demonstrate that the enzyme is localized exclusively in the cytoplasm at neutral pH, but is recruited to the membrane when the pH falls. We show by structure-based site-directed mutagenesis that the triple helix bundle formed by the N-termini of the protein at acidic pH is the major determinant for this behaviour. PMID- 12912903 TI - Channelling and formation of 'active' formaldehyde in dimethylglycine oxidase. AB - Here we report crystal structures of dimethylglycine oxidase (DMGO) from the bacterium Arthrobacter globiformis, a bifunctional enzyme that catalyzes the oxidation of N,N-dimethyl glycine and the formation of 5,10-methylene tetrahydrofolate. The N-terminal region binds FAD covalently and oxidizes dimethylglycine to a labile iminium intermediate. The C-terminal region binds tetrahydrofolate, comprises three domains arranged in a ring-like structure and is related to the T-protein of the glycine cleavage system. The complex with folinic acid indicates that this enzyme selectively activates the N10 amino group for initial attack on the substrate. Dead-end reactions with oxidized folate are avoided by the strict stereochemical constraints imposed by the folate-binding funnel. The active sites in DMGO are approximately 40 A apart, connected by a large irregular internal cavity. The tetrahydrofolate-binding funnel serves as a transient entry-exit port, and access to the internal cavity is controlled kinetically by tetrahydrofolate binding. The internal cavity enables sequestration of the reactive iminium intermediate prior to reaction with tetrahydrofolate and avoids formation of toxic formaldehyde. This mode of channelling in DMGO is distinct from other channelling mechanisms. PMID- 12912904 TI - Gain-of-function mutations indicate that Escherichia coli Kch forms a functional K+ conduit in vivo. AB - Although Kch of Escherichia coli is thought to be a K(+) channel by sequence homology, there is little evidence that it actually conducts K(+) ions in vitro or in vivo. We isolated gain-of-function (GOF) Kch mutations that render bacteria specifically sensitive to K(+) ions. Millimolar added K(+), but not Na(+) or sorbitol, blocks the initiation or continuation of mutant growth in liquid media. The mutations are mapped at the RCK (or KTN) domain, which is considered to be the cytoplasmic sensor controlling the gate. Additional mutations directed to the K(+)-filter sequence rescue the GOF mutant. The apparent K(+)-specific conduction through the 'loose-cannon' mutant channel suggests that the wild-type Kch channel also conducts, albeit in a regulated manner. Changing the internal ATG does not erase the GOF toxicity, but removes kch's short second product, suggesting that it is not required for channel function in vivo. The mutant phenotypes are better explained by a perturbation of membrane potential instead of internal K(+) concentration. Possible implications on the normal function of Kch are discussed. PMID- 12912905 TI - Activation of phospholipase D by the small GTPase Sar1p is required to support COPII assembly and ER export. AB - The small GTPase Sar1p controls the assembly of the cytosolic COPII coat that mediates export from the endoplasmic reticulum (ER). Here we demonstrate that phospholipase D (PLD) activation is required to support COPII-mediated ER export. PLD activity by itself does not lead to the recruitment of COPII to the membranes or ER export. However, PLD activity is required to support Sar1p-dependent membrane tubulation, the subsequent Sar1p-dependent recruitment of Sec23/24 and Sec13/31 COPII complexes to ER export sites and ER export. Sar1p recruitment to the membrane is PLD independent, yet activation of Sar1p is required to stimulate PLD activity on ER membranes, thus PLD is temporally regulated to support ER export. Regulated modification of membrane lipid composition is required to support the cooperative interactions that enable selective transport, as we demonstrate here for the mammalian COPII coat. PMID- 12912906 TI - GCNF-dependent repression of BMP-15 and GDF-9 mediates gamete regulation of female fertility. AB - To determine the function of germ cell nuclear factor (GCNF) in female reproduction, we generated an oocyte-specific GCNF knockout mouse model (GCNF(fl/fl)Zp3Cre(+)). These mice displayed hypofertility due to prolonged diestrus phase of the estrous cycle and aberrant steroidogenesis. These reproductive defects were secondary to a primary defect in the oocytes, in which expression of the paracrine transforming growth factor-beta signaling molecules, bone morphogenetic protein 15 (BMP-15) and growth differentiation factor 9 (GDF 9), were up-regulated in GCNF(fl/fl)Zp3Cre(+) females at diestrus. This was a direct effect of GCNF, as molecular studies showed that GCNF bound to DR0 elements within the BMP-15 and GDF-9 gene promoters and repressed their reporter activities. Consistent with these findings, abnormal double-oocyte follicles, indicative of aberrant BMP-15/GDF-9 expression, were observed in GCNF(fl/fl)Zp3Cre(+) females. The Cre/loxP knockout of GCNF in the oocyte has uncovered a new regulatory pathway in ovarian function. Our results show that GCNF directly regulates paracrine communication between the oocyte and somatic cells by regulating the expression of BMP-15 and GDF-9, to affect female fertility. PMID- 12912907 TI - HIF prolyl-hydroxylase 2 is the key oxygen sensor setting low steady-state levels of HIF-1alpha in normoxia. AB - Hypoxia-inducible factor (HIF), a transcriptional complex conserved from Caenorhabditis elegans to vertebrates, plays a pivotal role in cellular adaptation to low oxygen availability. In normoxia, the HIF-alpha subunits are targeted for destruction by prolyl hydroxylation, a specific modification that provides recognition for the E3 ubiquitin ligase complex containing the von Hippel-Lindau tumour suppressor protein (pVHL). Three HIF prolyl-hydroxylases (PHD1, 2 and 3) were identified recently in mammals and shown to hydroxylate HIF alpha subunits. Here we show that specific 'silencing' of PHD2 with short interfering RNAs is sufficient to stabilize and activate HIF-1alpha in normoxia in all the human cells investigated. 'Silencing' of PHD1 and PHD3 has no effect on the stability of HIF-1alpha either in normoxia or upon re-oxygenation of cells briefly exposed to hypoxia. We therefore conclude that, in vivo, PHDs have distinct assigned functions, PHD2 being the critical oxygen sensor setting the low steady-state levels of HIF-1alpha in normoxia. Interestingly, PHD2 is upregulated by hypoxia, providing an HIF-1-dependent auto-regulatory mechanism driven by the oxygen tension. PMID- 12912908 TI - Contrasting effects of VEGF gene disruption in embryonic stem cell-derived versus oncogene-induced tumors. AB - Previous gene targeting studies have implicated an indispensable role of vascular endothelial growth factor (VEGF) in tumor angiogenesis, particularly in tumors of embryonal or endocrine origin. In contrast, we report here that transformation of VEGF-deficient adult fibroblasts (MDF528) with ras or neu oncogenes gives rise to highly tumorigenic and angiogenic fibrosarcomas. These aggressive VEGF-null tumors (528ras, 528neu) originated from VEGF(-/-) embryonic stem cells, which themselves were tumorigenically deficient. We also report that VEGF production by tumor stroma has a modest role in oncogene-driven tumor angiogenesis. Both ras and neu oncogenes down-regulated at least two endogenous inhibitors of angiogenesis [pigment epithelium derived factor (PEDF) and thrombospondin 1 (TSP 1)]. This is functionally important as administration of an antiangiogenic TSP-1 peptide (ABT-526) markedly inhibited growth of VEGF(-/-) tumors, with some ingress of pericytes. These results provide the first definitive genetic demonstration of the dispensability of tumor cell-derived VEGF in certain cases of 'adult' tumor angiogenesis, and thus highlight the importance of considering VEGF-independent as well as VEGF-dependent pathways when attempting to block this process pharmacologically. PMID- 12912909 TI - A signalling role for 4-hydroxy-2-nonenal in regulation of mitochondrial uncoupling. AB - Oxidative stress and mitochondrial dysfunction are associated with disease and aging. Oxidative stress results from overproduction of reactive oxygen species (ROS), often leading to peroxidation of membrane phospholipids and production of reactive aldehydes, particularly 4-hydroxy-2-nonenal. Mild uncoupling of oxidative phosphorylation protects by decreasing mitochondrial ROS production. We find that hydroxynonenal and structurally related compounds (such as trans retinoic acid, trans-retinal and other 2-alkenals) specifically induce uncoupling of mitochondria through the uncoupling proteins UCP1, UCP2 and UCP3 and the adenine nucleotide translocase (ANT). Hydroxynonenal-induced uncoupling was inhibited by potent inhibitors of ANT (carboxyatractylate and bongkrekate) and UCP (GDP). The GDP-sensitive proton conductance induced by hydroxynonenal correlated with tissue expression of UCPs, appeared in yeast mitochondria expressing UCP1 and was absent in skeletal muscle mitochondria from UCP3 knockout mice. The carboxyatractylate-sensitive hydroxynonenal stimulation correlated with ANT content in mitochondria from Drosophila melanogaster expressing different amounts of ANT. Our findings indicate that hydroxynonenal is not merely toxic, but may be a biological signal to induce uncoupling through UCPs and ANT and thus decrease mitochondrial ROS production. PMID- 12912910 TI - Sequential recognition of two distinct sites in sigma(S) by the proteolytic targeting factor RssB and ClpX. AB - sigma(S) (RpoS), the master regulator of the general stress response in Escherichia coli, is a model system for regulated proteolysis in bacteria. sigma(S) turnover requires ClpXP and the response regulator RssB, whose phosphorylated form exhibits high affinity for sigma(S). Here, we demonstrate that recognition by the RssB/ClpXP system involves two distinct regions in sigma(S). Region 2.5 of sigma(S) (a long alpha-helix) is sufficient for binding of phosphorylated RssB. However, this interaction alone is not sufficient to trigger proteolysis. A second region located in the N-terminal part of sigma(S), which is exposed only upon RssB-sigma(S) interaction, serves as a binding site for the ClpX chaperone. Binding of the ClpX hexameric ring to sigma(S)-derived reporter proteins carrying the ClpX-binding site (but not the RssB-binding site) is also not sufficient to commit the protein to degradation. Our data indicate that RssB plays a second role in the initiation of sigma(S) proteolysis that goes beyond targeting of sigma(S) to ClpX, and suggest a model for the sequence of events in the initiation of sigma(S) proteolysis. PMID- 12912911 TI - Receptor protein tyrosine phosphatase alpha is essential for hippocampal neuronal migration and long-term potentiation. AB - Despite clear indications of their importance in lower organisms, the contributions of protein tyrosine phosphatases (PTPs) to development or function of the mammalian nervous system have been poorly explored. In vitro studies have indicated that receptor protein tyrosine phosphatase alpha (RPTPalpha) regulates SRC family kinases, potassium channels and NMDA receptors. Here, we report that absence of RPTPalpha compromises correct positioning of pyramidal neurons during development of mouse hippocampus. Thus, RPTPalpha is a novel member of the functional class of genes that control radial neuronal migration. The migratory abnormality likely results from a radial glial dysfunction rather than from a neuron-autonomous defect. In spite of this aberrant development, basic synaptic transmission from the Schaffer collateral pathway to CA1 pyramidal neurons remains intact in Ptpra(-/-) mice. However, these synapses are unable to undergo long-term potentiation. Mice lacking RPTPalpha also underperform in the radial arm water-maze test. These studies identify RPTPalpha as a key mediator of neuronal migration and synaptic plasticity. PMID- 12912913 TI - Focal adhesion kinase signaling promotes phagocytosis of integrin-bound photoreceptors. AB - Daily alphavbeta5 integrin-dependent phagocytosis of spent photoreceptor outer segment fragments by the retinal pigment epithelium (RPE) is critical for retinal function. This study identifies a key role for focal adhesion kinase (FAK) in RPE phagocytosis. Particle binding increases FAK complex formation with alphavbeta5 receptors at the apical, phagocytic RPE surface and activates FAK. Subsequent particle engulfment coincides with dissociation of activated FAK from alphavbeta5. Mutant FAK retaining focal adhesion targeting but lacking kinase activity interferes with recruitment of full-length FAK to alphavbeta5 and abrogates FAK activation in response to RPE phagocytic challenge. Such inhibition of FAK signaling has no effect on alphavbeta5-dependent binding of particles but blocks their engulfment. Conversely, FAK re-expression promotes particle engulfment by FAK null fibroblasts. Selective ligation of alphavbeta5 receptors at the apical RPE surface is sufficient to phosphorylate and mobilize FAK. Furthermore, FAK phagocytic signaling is independent of the internalization receptor MerTK. In contrast, inhibition of FAK signaling diminishes MerTK phosphorylation. These results demonstrate that FAK provides an essential link between binding and engulfment mechanisms of integrin-mediated phagocytosis. PMID- 12912912 TI - Interdimer processing mechanism of procaspase-8 activation. AB - The execution of apoptosis depends on the hierarchical activation of caspases. The initiator procaspases become autoproteolytically activated through a less understood process that is triggered by oligomerization. Procaspase-8, an initiator caspase recruited to death receptors, is activated through two cleavage events that proceed in a defined order to generate the large and small subunits of the mature protease. Here we show that dimerization of procaspase-8 produces enzymatically competent precursors through the stable homophilic interaction of the procaspase-8 protease domain. These dimers are also more susceptible to processing than individual procaspase-8 molecules, which leads to their cross cleavage. The order of the two interdimer cleavage events is maintained by a sequential accessibility mechanism: the separation of the large and small subunits renders the region between the large subunit and prodomain susceptible to further cleavage. In addition, the activation process involves an alteration in the enzymatic properties of caspase-8; while procaspase-8 molecules specifically process one another, mature caspases only cleave effector caspases. These results reveal the key steps leading to the activation of procaspase-8 by oligomerization. PMID- 12912914 TI - Pak1 and PIX regulate contact inhibition during epithelial wound healing. AB - Wound healing in epithelia requires coordinated cell migration and proliferation regulated by signaling mechanisms that are poorly understood. Here we show that epithelial cells expressing constitutively active or kinase-dead mutants of the Rac/Cdc42 effector Pak1 fail to undergo growth arrest upon wound closure. Strikingly, this phenotype is only observed when the Pak1 kinase mutants are expressed in cells possessing a free lateral surface, i.e. one that is not engaged in contact with neighboring cells. The Pak1 kinase mutants perturb contact inhibition by a mechanism that depends on the Pak-interacting Rac-GEF PIX. In control cells, endogenous activated Pak and PIX translocate from focal complexes to cell-cell contacts during wound closure. This process is abrogated in cells expressing Pak1 kinase mutants. In contrast, Pak1 mutants rendered defective in PIX binding do not impede translocation of activated Pak and PIX, and exhibit normal wound healing. Thus, recruitment of activated Pak and PIX to cell-cell contacts is pivotal to transduction of growth-inhibitory signals from neighboring cells in epithelial wound healing. PMID- 12912915 TI - Differential regulation of NFAT and SRF by the B cell receptor via a PLCgamma Ca(2+)-dependent pathway. AB - NFAT and SRF are important in the regulation of proliferation and cytokine production in lymphocytes. NFAT activation by the B cell receptor (BCR) occurs via the PLCgamma-Ca(2+)-calcineurin pathway, however how the BCR activates SRF is unclear. We show here that like NFAT, BCR regulation of SRF occurs via an Src-Syk Tec-PLCgamma-Ca(2+) (Lyn-Syk-Btk-PLCgamma-Ca(2+)) pathway. However, SRF responds to lower Ca(2+) and is less dependent on IP(3)R expression than NFAT. Ca(2+) regulated calcineurin plays a partial role in SRF activation, in combination with diacylglycerol (DAG), while is fully required for NFAT activation. Signals from the DAG effectors protein kinase C, Ras and Rap1, and the downstream MEK-ERK pathway are required for both SRF and NFAT; however, NFAT but not SRF is dependent on JNK signals. Both SRF and NFAT were also dependent on Rac, Rho, CDC42 and actin. Finally, we show that Ca(2+) is not required for ERK activation, but instead for its association with nuclear areas of the cell. These data suggest that combinatorial assembly of signaling pathways emanating from the BCR differentially regulate NFAT and SRF, to activate gene expression. PMID- 12912916 TI - Insulin induces phosphatidylinositol-3-phosphate formation through TC10 activation. AB - Phosphatidylinositol-3-phosphate (PtdIns-3-P) is considered as a lipid constitutively present on endosomes; it does not seem to have a dynamic role in signalling. In contrast, phosphatidylinositol-3,4,5-trisphosphate (PtdIns-3,4,5 P(3)) plays a crucial role in different signalling pathways including translocation of the glucose transporter protein GLUT4 to the plasma membrane upon insulin receptor activation. GLUT4 translocation requires activation of two distinct pathways involving phosphatidylinositol 3-kinase (PI 3-K) and the small GTP-binding protein TC10, respectively. The contribution of each pathway remains to be elucidated. Here we show that insulin specifically induces the formation of PtdIns-3-P in insulin- responsive cells. The insulin-mediated formation of PtdIns 3-P occurs through the activation of TC10 at the lipid rafts subdomain of the plasma membrane. Exogenous PtdIns-3-P induces the plasma membrane translocation of both overexpressed and endogenous GLUT4. These data indicate that PtdIns-3-P is specifically produced downstream from insulin-mediated activation of TC10 to promote the plasma membrane translocation of GLUT4. These results give a new insight into the intracellular role of PtdIns-3-P and shed light on some aspects of insulin signalling so far not completely understood. PMID- 12912917 TI - The in vivo roles of STEF/Tiam1, Rac1 and JNK in cortical neuronal migration. AB - The coordinated migration of neurons is a pivotal step for functional architectural formation of the mammalian brain. To elucidate its molecular mechanism, gene transfer by means of in utero electroporation was applied in the developing murine brain, revealing the crucial roles of Rac1, its activators, STEF/Tiam1, and its downstream molecule, c-Jun N-terminal kinase (JNK), in the cerebral cortex. Functional repression of these molecules resulted in inhibition of radial migration of neurons without affecting their proper differentiation. Interestingly, distinct morphological phenotypes were observed; suppression of Rac1 activity caused loss of the leading process, whereas repression of JNK activity did not, suggesting the complexity of the signaling cascade. In cultured neurons from the intermediate zone, activated JNK was detected along microtubules in the processes. Application of a JNK inhibitor caused irregular morphology and increased stable microtubules in processes, and decreased phosphorylation of microtubule associated protein 1B, raising a possibility of the involvement of JNK in controlling tubulin dynamics in migrating neurons. Our data thus provide important clues for understanding the intracellullar signaling machinery for cortical neuronal migration. PMID- 12912918 TI - In vivo role of the PIF-binding docking site of PDK1 defined by knock-in mutation. AB - PKB/Akt, S6K, SGK and RSK are mediators of responses triggered by insulin and growth factors and are activated following phosphorylation by 3-phosphoinositide dependent protein kinase-1 (PDK1). To investigate the importance of a substrate docking site in the kinase domain of PDK1 termed the 'PIF-pocket', we generated embryonic stem (ES) cells in which both copies of the PDK1 gene were altered by knock-in mutation to express a form of PDK1 retaining catalytic activity, in which the PIF-pocket site was disrupted. The knock-in ES cells were viable, mutant PDK1 was expressed at normal levels and insulin-like growth factor 1 induced normal activation of PKB and phosphorylation of the PKB substrates GSK3 and FKHR. In contrast, S6K, RSK and SGK were not activated, nor were physiological substrates of S6K and RSK phosphorylated. These experiments establish the importance of the PIF-pocket in governing the activation of S6K, RSK, SGK, but not PKB, in vivo. They also illustrate the power of knock-in technology to probe the physiological roles of docking interactions in regulating the specificity of signal transduction pathways. PMID- 12912919 TI - Reversal of human cellular senescence: roles of the p53 and p16 pathways. AB - Telomere erosion and subsequent dysfunction limits the proliferation of normal human cells by a process termed replicative senescence. Replicative senescence is thought to suppress tumorigenesis by establishing an essentially irreversible growth arrest that requires activities of the p53 and pRB tumor suppressor proteins. We show that, depending on expression of the pRB regulator p16, replicative senescence is not necessarily irreversible. We used lentiviruses to express specific viral and cellular proteins in senescent human fibroblasts and mammary epithelial cells. Expression of telomerase did not reverse the senescence arrest. However, cells with low levels of p16 at senescence resumed robust growth upon p53 inactivation, and limited growth upon expression of oncogenic RAS. In contrast, cells with high levels of p16 at senescence failed to proliferate upon p53 inactivation or RAS expression, although they re-entered the cell cycle without growth after pRB inactivation. Our results indicate that the senescence response to telomere dysfunction is reversible and is maintained primarily by p53. However, p16 provides a dominant second barrier to the unlimited growth of human cells. PMID- 12912920 TI - Regulation of PI4,5P2 synthesis by nuclear-cytoplasmic shuttling of the Mss4 lipid kinase. AB - The essential phospholipid PI4,5P(2) is generated by a well conserved PI4P 5 kinase, Mss4, in yeast. Balanced production and turnover of PI4,5P(2) is important for normal organization of the actin cytoskeleton and cell viability. Previous studies have shown that multiple PI phosphatases can regulate PI4,5P(2) levels. We report a new, unexpected regulatory mechanism for PI4,5P(2) homeostasis, directed by nuclear-cytoplasmic shuttling of the lipid kinase. We show that Mss4 is a phosphoprotein, which contains a functional nuclear localization signal (NLS) and can shuttle between the cytoplasm and the nucleus. Temperature-conditional mss4 cells that accumulate Mss4 protein in the nucleus exhibit reduced levels of PI4,5P(2), depolarization of the actin cytoskeleton and a block in Mss4 phosphorylation, suggesting an essential role for phosphorylated Mss4 at the plasma membrane. Through the isolation of gene dosage-dependent suppressors of mss4 mutants, we identified Bcp1, a protein enriched in the nucleus, which is required for Mss4 nuclear export and is related to the mammalian BRCA2-interacting protein BCCIP. Together, these studies suggest a new mechanism for lipid kinase regulation through regulated nuclear-cytoplasmic shuttling. PMID- 12912922 TI - Identification of the RNA polymerase II subunit hsRPB7 as a novel target of the von Hippel-Lindau protein. AB - Inactivation of the von Hippel-Lindau (VHL) tumor suppressor gene is linked to the hereditary VHL disease and sporadic clear cell renal cell carcinomas (CCRCC). VHL-associated tumors are highly vascularized, a characteristic associated with overproduction of vascular endothelial growth factor (VEGF). The VHL protein (pVHL) is a component of the ubiquitin ligase E3 complex, targeting substrate proteins for ubiquitylation and subsequent proteasomic degradation. Here, we report that the pVHL can directly bind to the human RNA polymerase II seventh subunit (hsRPB7) through its beta-domain, and naturally occurring beta-domain mutations can decrease the binding of pVHL to hsRPB7. Introducing wild-type pVHL into human kidney tumor cell lines carrying endogenous mutant non-functional pVHL facilitates ubiquitylation and proteasomal degradation of hsRPB7, and decreases its nuclear accumulation. pVHL can also suppress hsRPB7-induced VEGF promoter transactivation, mRNA expression and VEGF protein secretion. Together, our results suggest that hsRPB7 is a downstream target of the VHL ubiquitylating complex and pVHL may regulate angiogenesis by targeting hsRPB7 for degradation via the ubiquitylation pathway and preventing VEGF expression. PMID- 12912921 TI - Distinct requirements for the naturally occurring splice forms Stat4alpha and Stat4beta in IL-12 responses. AB - Signal transducer and activator of transcription (Stat)4 is a signaling molecule required for normal responses to interleukin-12 (IL-12) and is critically involved in inflammatory responses. We have isolated an alternatively spliced isoform of Stat4, termed Stat4beta, which lacks 44 amino acids at the C-terminus, encompassing the putative transcriptional activation domain. To assess the in vivo roles of these Stat4 isoforms, we generated transgenic Stat4-deficient mice expressing Stat4alpha or Stat4beta. Our results indicate that T-cell-specific expression of Stat4alpha or Stat4beta can mediate many aspects of IL-12 signaling including the differentiation of Th1 cells. However, Stat4alpha is required for normal levels of IL-12-induced interferon-gamma production from Th1 cells. Microarray analysis identified 98 genes induced by both Stat4 isoforms, 32 genes induced only by Stat4alpha and 29 genes induced only by Stat4beta. Some induced genes correlate with specific functions including the ability of Stat4beta, but not Stat4alpha, to mediate IL-12-stimulated proliferation. Thus, Stat4alpha and Stat4beta have distinct roles in mediating responses to IL-12. PMID- 12912923 TI - GATA-6 maintains BMP-4 and Nkx2 expression during cardiomyocyte precursor maturation. AB - GATA-6 is expressed in presumptive cardiac mesoderm before gastrulation, but its role in heart development has been unclear. Here we show that Xenopus and zebrafish embryos, injected with antisense morpholino oligonucleotides designed specifically to knock-down translation of GATA-6 protein, are severely compromised for heart development. Injected embryos express greatly reduced levels of contractile machinery genes and, at the same stage, of regulatory genes such as bone morphogenetic protein-4 (BMP-4) and the Nkx2 family. In contrast, initial BMP and Nkx2 expression is normal, suggesting a maintenance role for GATA 6. Endoderm is critical for heart formation in several vertebrates including Xenopus, and separate perturbation of GATA-6 expression in the deep anterior endoderm and in the overlying heart mesoderm shows that GATA-6 is required in both for cardiogenesis. The GATA-6 requirement in cardiac mesoderm was confirmed in zebrafish, an organism in which endoderm is thought not to be necessary for heart formation. We therefore conclude that proper maturation of cardiac mesoderm requires GATA-6, which functions to maintain BMP-4 and Nkx2 expression. PMID- 12912924 TI - A 10 residue motif at the C-terminus of the RNA pol II CTD is required for transcription, splicing and 3' end processing. AB - The RNA polymerase II C-terminal heptad repeat domain (CTD) is essential for normal transcription and co-transcriptional processing of mRNA precursors. The mammalian CTD comprises 52 heptads whose consensus, YSPTSPS, is conserved throughout eukaryotes, followed by a 10 amino acid C-terminal sequence that is conserved only among vertebrates. Here we show that surprisingly, the heptad repeats are not sufficient to support efficient transcription, pre-mRNA processing or full cell viability. In addition to the heptads, the 10 amino acid C-terminal motif is essential for high level transcription, splicing and poly(A) site cleavage. Efficient mRNA synthesis from a transiently transfected reporter gene required the C-terminal motif plus between 16 and 25 heptad repeats from either the N- or C-terminal half of the CTD. Twenty-seven consensus heptads plus the C-terminal motif also supported efficient mRNA synthesis but not cell viability. PMID- 12912925 TI - A common sequence motif determines the Cajal body-specific localization of box H/ACA scaRNAs. AB - Post-transcriptional synthesis of 2'-O-methylated nucleotides and pseudouridines in Sm spliceosomal small nuclear RNAs takes place in the nucleoplasmic Cajal bodies and it is directed by guide RNAs (scaRNAs) that are structurally and functionally indistinguishable from small nucleolar RNAs (snoRNAs) directing rRNA modification in the nucleolus. The scaRNAs are synthesized in the nucleoplasm and specifically targeted to Cajal bodies. Here, mutational analysis of the human U85 box C/D-H/ACA scaRNA, followed by in situ localization, demonstrates that box H/ACA scaRNAs share a common Cajal body-specific localization signal, the CAB box. Two copies of the evolutionarily conserved CAB consensus (UGAG) are located in the terminal loops of the 5' and 3' hairpins of the box H/ACA domains of mammalian, Drosophila and plant scaRNAs. Upon alteration of the CAB boxes, mutant scaRNAs accumulate in the nucleolus. In turn, authentic snoRNAs can be targeted into Cajal bodies by addition of exogenous CAB box motifs. Our results indicate that scaRNAs represent an ancient group of small nuclear RNAs which are localized to Cajal bodies by an evolutionarily conserved mechanism. PMID- 12912926 TI - Localization of proteins bound to a replication origin of human DNA along the cell cycle. AB - The proteins bound in vivo at the human lamin B2 DNA replication origin and their precise sites of binding were investigated along the cell cycle utilizing two novel procedures based on immunoprecipitation following UV irradiation with a pulsed laser light source. In G(1), the pre-replicative complex contains CDC6, MCM3, ORC1 and ORC2 proteins; of these, the post-replicative complex in S phase contains only ORC2; in M phase none of them are bound. The precise nucleotide of binding was identified for the two ORC and the CDC6 proteins near the start sites for leading-strand synthesis; the transition from the pre- to the post replicative complex is accompanied by a 17 bp displacement of the ORC2 protein towards the start site. PMID- 12912927 TI - Elg1 forms an alternative RFC complex important for DNA replication and genome integrity. AB - Genome-wide synthetic genetic interaction screens with mutants in the mus81 and mms4 replication fork-processing genes identified a novel replication factor C (RFC) homolog, Elg1, which forms an alternative RFC complex with Rfc2-5. This complex is distinct from the DNA replication RFC, the DNA damage checkpoint RFC and the sister chromatid cohesion RFC. As expected from its genetic interactions, elg1 mutants are sensitive to DNA damage. Elg1 is redundant with Rad24 in the DNA damage response and contributes to activation of the checkpoint kinase Rad53. We find that elg1 mutants display DNA replication defects and genome instability, including increased recombination and mutation frequencies, and minichromosome maintenance defects. Mutants in elg1 show genetic interactions with pathways required for processing of stalled replication forks, and are defective in recovery from DNA damage during S phase. We propose that Elg1-RFC functions both in normal DNA replication and in the DNA damage response. PMID- 12912928 TI - Nucleotide shuffling and ssDNA recognition in Oxytricha nova telomere end-binding protein complexes. AB - Sequence-specific protein recognition of single-stranded nucleic acids is critical for many fundamental cellular processes, such as DNA replication, DNA repair, transcription, translation, recombination, apoptosis and telomere maintenance. To explore the mechanisms of sequence-specific ssDNA recognition, we determined the crystal structures of 10 different non-cognate ssDNAs complexed with the Oxytricha nova telomere end-binding protein (OnTEBP) and evaluated their corresponding binding affinities (PDB ID codes 1PH1-1PH9 and 1PHJ). The thermodynamic and structural effects of these sequence perturbations could not have been predicted based solely upon the cognate structure. OnTEBP accommodates non-cognate nucleotides by both subtle adjustments and surprisingly large structural rearrangements in the ssDNA. In two complexes containing ssDNA intermediates that occur during telomere extension by telomerase, entire nucleotides are expelled from the complex. Concurrently, the sequence register of the ssDNA shifts to re-establish a more cognate-like pattern. This phenomenon, termed nucleotide shuffling, may be of general importance in protein recognition of single-stranded nucleic acids. This set of structural and thermodynamic data highlights a fundamental difference between protein recognition of ssDNA versus dsDNA. PMID- 12912930 TI - Methylation and prognosis: of molecular clocks and hypermethylator phenotypes. PMID- 12912929 TI - DNA polymerase stabilization at stalled replication forks requires Mec1 and the RecQ helicase Sgs1. AB - To ensure proper replication and segregation of the genome, eukaryotic cells have evolved surveillance systems that monitor and react to impaired replication fork progression. In budding yeast, the intra-S phase checkpoint responds to stalled replication forks by downregulating late-firing origins, preventing spindle elongation and allowing efficient resumption of DNA synthesis after recovery from stress. Mutations in this pathway lead to high levels of genomic instability, particularly in the presence of DNA damage. Here we demonstrate by chromatin immunoprecipitation that when yeast replication forks stall due to hydroxyurea (HU) treatment, DNA polymerases alpha and epsilon are stabilized for 40-60 min. This requires the activities of Sgs1, a member of the RecQ family of DNA helicases, and the ATM-related kinase Mec1, but not Rad53 activation. A model is proposed whereby Sgs1 helicase resolves aberrantly paired structures at stalled forks to maintain single-stranded DNA that allows RP-A and Mec1 to promote DNA polymerase association. PMID- 12912931 TI - Immunophilins, mTOR, and pharmacodynamic strategies for a targeted cancer therapy. PMID- 12912932 TI - Pharmacodynamic Evaluation of CCI-779, an Inhibitor of mTOR, in Cancer Patients. AB - CCI-779 is an ester of rapamycin and inhibitor of mammalian target of rapamycin (mTOR) currently in Phase II clinical development for the treatment of patients with cancer. CCI-779 interacts with mTOR and inhibits its kinase activity, resulting in inhibition of the mTOR-regulated translational controllers p70(s6) kinase and 4E-BP1. Ultimately, CCI-779 decreases the translation of mRNAs involved in the control of the cell cycle, resulting in cell cycle arrest. The objective of this study was to develop a method to determine the pharmacodynamic effects of CCI-779 suitable for use in clinical trials. Exposure of Raji lymphoblastoid cells to increasing concentrations of rapamycin resulted in a linear concentration-dependent inhibition of p70(s6) kinase activity, suggesting that p70(s6) kinase activity could be an appropriate marker for mTOR-interacting agents. In subsequent experiments, treatment of nude mice bearing the CCI-779 susceptible breast cancer cell line MDA-468 with a single dose of 10 mg/kg CCI 779 resulted in a >80% inhibition of p70(s6) kinase activity in peripheral blood mononuclear cells (PBMCs) 72 h after treatment. Importantly, the degree of p70(s6) kinase inhibition was identical in PBMCs and simultaneously collected tumor tissue, suggesting that the PBMCs are an adequate surrogate tissue for p70(s6) kinase activity in vivo. The intrasubject coefficient of variation of p70(s6) kinase activity measured in PBMCs collected from five healthy volunteers on days 1, 4, and 8 was 14%, indicating that p70(s6) kinase activity in PBMCs remains relatively stable over time. Finally, p70(s6) kinase activity was measured in PBMCs from nine patients with renal cell cancer treated with a single dose of 25, 75, or 250 mg of CCI-779 i.v. (three patients each). PBMCs were collected on days 2, 4, and 8 after CCI-779 treatment. In this small data set, eight of the nine patients had evidence of p70(s6) kinase activity inhibition after treatment that was independent of the administered dose. There was a significant linear association between time to disease progression and inhibition of p70(s6) kinase activity 24 h after treatment. In conclusion, these results indicate that the pharmacodynamic effects of CCI-779 can be determined using a p70(s6) kinase assay in PBMCs. This assay is currently being incorporated in Phase I and II studies with CCI-779 to determine its relationship with dose and plasma concentration of the agent and its value as a predictor of treatment efficacy. PMID- 12912933 TI - Zoledronic acid induces significant and long-lasting modifications of circulating angiogenic factors in cancer patients. AB - PURPOSE: The commercial availability of zoledronic acid, a third generation bisphosphonate, prompted us to evaluate the modifications in angiogenic cytokines levels after a single i.v. infusion of this drug. EXPERIMENTAL DESIGN: Thirty consecutive cancer patients with scintigraphic and radiographic evidence of bone metastases were treated with a single infusion of 4 mg of zoledronic acid before any chemotherapy. The patients were prospectively evaluated for circulating levels of vascular endothelial growth factor (VEGF) and platelet-derived growth factor (PDGF) just before and at 1, 2, 7, and 21 days after zoledronic acid infusion. RESULTS: Basal serum VEGF median levels were significantly decreased at days 2 (-23%), 7 (-28%), and 21 (-34%) after zoledronic acid infusion (P = 0.0498, 0.0090, and 0.0011, respectively). Serum PDGF levels were significantly decreased by 25% 1 day after zoledronic acid infusion (P = 0.0032). This effect on circulating PDGF levels persisted for 2 days after bisphosphonate infusion (P = 0.0050). PDGF levels had returned to values similar to the median basal value at 7 and 21 days. Moreover, a linear regression model with variance analysis showed a significant positive correlation between basal VEGF and PDGF values but not at the following time points. No significant differences were recorded in platelet levels, WBC count, or hemoglobin concentration before and after zoledronic acid single infusion. CONCLUSIONS: This study confirms that zoledronic acid could have an in vivo antiangiogenic property through a significant and long lasting reduction in serum VEGF levels. PMID- 12912934 TI - CpG island methylator phenotype is an independent predictor of survival benefit from 5-fluorouracil in stage III colorectal cancer. AB - PURPOSE: The CpG island methylator phenotype (CIMP) is observed in approximately 30% of colorectal cancer (CRC) cases and is characterized by the concurrent methylation of multiple CpG islands in tumor DNA. This phenotype (CIMP+) is more frequently observed in tumors with proximal location, microsatellite instability, and normal p53. Because it has previously been observed that each of these features is associated with a good survival benefit from 5-fluorouracil (5-FU) based adjuvant chemotherapy, we investigated in the present study whether CIMP+ has independent predictive value. EXPERIMENTAL DESIGN: CIMP+ status was evaluated in 103 stage III CRCs from patients treated with surgery alone and for an additional 103 cases from patients treated with surgery and adjuvant 5-FU-based chemotherapy. The two cohorts were randomly pair-matched for age, sex, and tumor site, and the median length of follow-up time was 39 months. RESULTS: CIMP+ status predicted survival benefit from 5-FU treatment independently of microsatellite instability and p53 mutation status (relative risk = 0.22; 95% confidence interval, 0.06-0.84; P = 0.027). Unmeasured, high-risk confounding factors could only account for this association if they were unequally distributed between the two patient cohorts by a factor of at least 2-fold. CONCLUSIONS: CIMP+ has independent predictive significance for the survival benefit from 5-FU chemotherapy in CRC. This molecular marker should be incorporated into prospective clinical trials of fluorouracil-based therapies to confirm its clinical value. PMID- 12912936 TI - Prognostic importance of promoter hypermethylation of multiple genes in esophageal adenocarcinoma. AB - PURPOSE: We investigated aberrant methylation patterns in esophageal adenocarcinoma and correlated the findings to patient survival and tumor recurrence. EXPERIMENTAL DESIGN: Gene promoter methylation was performed in 82 samples from 41 esophagectomy patients consisting of 41 adenocarcinoma samples, each with its adjacent nonmalignant tissue, which included one sample with Barretts metaplasia. The methylation status of seven genes was determined. Epigenetic silencing was confirmed using immunohistochemical staining. Kaplan Meier plots were constructed using disease-specific survival as the primary end point and the interval from surgery to tumor recurrence as the secondary end point. The association of clinicopathological and biomolecular risk factors to survival and recurrence was performed using the Log-rank test and Cox proportional hazards model for multivariate analysis. RESULTS: Methylation frequencies of the genes analyzed were APC, 68%; E-cadherin, 66%; O(6) methylguanine DNA methyltransferase, 56%; ER, 51%; p16, 39%; DAP-kinase, 19%; and TIMP3, 19%. DNA methylation of some genes individually showed only trends toward diminished survival, whereas patients whose tumors had >50% of their gene profile methylated had both significantly poorer survival (P = 0.04) and earlier tumor recurrence (P = 0.05) than those without positive methylation. By multivariate analysis, the hazard ratios (HRs) with positive methylation status were more powerful predictors of survival [HR 2.7 (1.14-6.45; 95% confidence interval)] and tumor recurrence [HR 2.5 (1.11-5.6)] than age (HR 2.03 and 1.96, respectively) or stage (HR 1.48 and 1.67, respectively). CONCLUSIONS: Our data suggest that positive methylation status for multiple genes in esophageal adenocarcinoma is a predictor of poor prognosis. PMID- 12912935 TI - Haptoglobin-alpha subunit as potential serum biomarker in ovarian cancer: identification and characterization using proteomic profiling and mass spectrometry. AB - PURPOSE: The objective of this study was to identify and characterize new serum biomarkers in ovarian cancer patients using mass spectrometric protein profiling and specific immunological assays. EXPERIMENTAL DESIGN: Serum samples from 80 cancer patients and 91 healthy women were analyzed by surface enhanced laser desorption and ionization-mass spectrometry (MS) profiling. A candidate biomarker was purified by affinity chromatography, and its sequence was determined by liquid chromatography-tandem MS. An antibody was generated from the synthesized peptide for quantitative validation in the cases and controls. CA125 was determined and compared with the same set of specimens. RESULTS: Using surface enhanced laser desorption and ionization, we found a serum biomarker at approximately 11700 Da, which had peak intensity significantly higher in cases (1.366) compared with controls (0.208, P = 0.002), and subsequently identified this as the alpha chain of haptoglobin. ELISA indicated that Hp-alpha was 80% of tumor cells stained positively. Five of 23 (22%) spontaneous soft tissue sarcomas of comparable histological types, including 2 angiosarcomas, were KIT positive. Molecular genetic analysis of exon 11 of the c-kit gene was attainable for 13 of the 16 postradiation sarcomas. No mutations were found. CONCLUSIONS: Postradiation sarcomas are aggressive malignancies, seldom amenable to curative treatment. A majority of the analyzed tumors showed extensive expression of the KIT protein, but no mutations in exon 11 of the c-kit gene were found. Still, without the availability of effective therapies, treatment with the KIT inhibitor imatinib mesylate might be considered for patients with postradiation sarcomas. PMID- 12912939 TI - Expression of constitutively active Akt-3 in MCF-7 breast cancer cells reverses the estrogen and tamoxifen responsivity of these cells in vivo. AB - PURPOSE: Prior studies had suggested that Akt activity is elevated in a subset of breast cancers. In this study, to test the effect of active Akt-3 on estrogen receptor function, we have produced MCF-7 cells, which express active Akt-3 and examined the estrogen responsiveness of these cells in vivo and in vitro. EXPERIMENTAL DESIGN: MCF-7 cells expressing active Akt-3 were studied for estradiol (E2) responsiveness in vitro by both using an estrogen receptor element reporter construct as well as looking at induction of endogenous genes. These cells were also studied in vivo after injection into nude, ovariectomized mice by following tumor growth rates in the presence or absence of E2, tamoxifen, or the pure antiestrogen, ICI 182,780 (fulvestrant). RESULTS: Akt-3-expressing cells were found to produce tumors in mice in the absence of E2 that were approximately equivalent in size to control cells in mice given E2. Moreover, the formation of tumors by the Akt-3 cells was greatly suppressed by E2, stimulated by tamoxifen, and unaffected by ICI 182,780. In the in vitro assays for gene induction by E2, the Akt-3-expressing cells exhibited similar E2 and tamoxifen responsiveness as the control cells. CONCLUSIONS: These results indicate that expression of active Akt-3 in MCF-7 cells results in E2-independent tumor growth. Moreover, the growth of these tumors is inhibited by E2 and enhanced by tamoxifen. Finally, these tumors are resistant to ICI 182,780. These findings suggest that the amount of active Akt present in breast cancers may be important in the relative efficacy of different treatments. PMID- 12912940 TI - Phase I clinical and pharmacokinetic study of irinotecan in adults with recurrent malignant glioma. AB - PURPOSE: A preliminary evaluation of the efficacy of irinotecan in patients with malignant glioma demonstrated modest activity. A markedly lower than expected incidence of drug-related toxicity was also noted. This was consistent with pharmacokinetic data indicating that the total body clearance (CL) of irinotecan in this patient population was considerably greater than in colorectal cancer patients. Concomitant medications used chronically in brain cancer patients, especially glucocorticoids and anticonvulsants that induce hepatic enzymes involved in the metabolism or excretion of drugs, were believed to be the cause of the alteration in pharmacokinetic behavior. A Phase I study was therefore undertaken in patients with recurrent malignant gliomas to independently determine the maximum tolerated dose (MTD) of irinotecan in patients stratified according to the use of enzyme-inducing anticonvulsants (EIAs). EXPERIMENTAL DESIGN: Patients with recurrent malignant gliomas received irinotecan as a weekly 90-min i.v. infusion for four consecutive weeks, with additional cycles of treatment repeated every 6 weeks. The starting dose was 125 mg/m(2)/week for both groups of patients (+/-EIA). Groups of >/==" BORDER="0">3 patients were evaluated at each dose level, and the modified continual reassessment method was used for dose adjustments. The plasma pharmacokinetics of irinotecan, its active metabolite, 7-ethyl-10-hydroxy-camptothecin (SN-38), and the glucuronide conjugate of SN-38, SN-38 glucuronide, were determined in all patients during treatment with the first weekly dose. RESULTS: Forty patients were enrolled into the study and treated with a total of 135 cycles of irinotecan. The MTD was determined to be 411 mg/m(2)/week in the +EIA cohort and 117 mg/m(2)/week in the EIA cohort for the weekly x 4 every 6 weeks schedule. Pharmacokinetic studies showed that the CL of irinotecan was distinctly dose dependent in the patients receiving EIAs, decreasing from approximately 50 liters/h/m(2) at the lower dose levels (125-238 mg/m(2)) to a mean +/- SD value of 29.7 +/- 9.0 liters/h/m(2) (n = 7) at the MTD. The grand mean CL for a group of 13 patients who were not taking EIAs, 18.8 +/- 10.6 liters/h/m(2), was significantly different from the mean CL at the MTD of the +EIA cohort (P = 0.033). Mean values of the AUC of SN-38 (P = 0.4) and SN-38 glucuronide (P = 0.55) were not significantly different at the MTDs for the two cohorts of patients. CONCLUSIONS: The MTD of irinotecan was 3.5 times greater in patients with malignant glioma who were concurrently receiving EIAs than in those who were not. This study has also served to confirm that the concomitant administration of EIAs results in marked enhancement in the CL of irinotecan. These findings have important implications for subsequent clinical trials to further evaluate irinotecan in brain cancer patients and underscore the importance of assessing the potential for pharmacokinetic interactions between concurrent medications and chemotherapeutic agents. PMID- 12912941 TI - Pharmacokinetics and immunological aspects of a phase Ib study with intratumoral administration of recombinant human interleukin-12 in patients with head and neck squamous cell carcinoma: a decrease of T-bet in peripheral blood mononuclear cells. AB - The aim of this study was to evaluate the tolerability of intratumoral administered recombinant human interleukin-12 (rhIL-12) in patients with head and neck squamous cell carcinoma. Six patients were treated once a week at two dose levels of 100 or 300 ng/kg, respectively, up to 24 weeks. The primary end point was to assess the toxicity and safety of intratumoral injected rhIL-12 in head and neck squamous cell carcinoma patients; the pharmacokinetics and pharmacodynamics of rhIL-12 and any evidence of antitumor effect were also determined. Toxicity was mild, with prolonged grade 4 lymphopenia observed in only one patient. No dose-limiting toxicities occurred. In all six patients, the rhIL-12 was detectable in plasma within 30 min. Significant reductions in absolute number of peripheral blood lymphocytes and all lymphocyte subsets, especially cytotoxic T cells and natural killer cells, were observed that were maximal between 12 and 24 h. Maximal plasma concentrations of IFN-gamma and IL-10 were detected after 12 h. A real-time semiquantitative PCR analysis in peripheral blood mononuclear cells showed a mean increase of mRNA encoding IFN-gamma of 2.2 times relative to the pretreatment sample. An unexpected, significant decrease of 80% in T-bet mRNA, a T-helper 1 transcription factor, was detected after 12 h, with normalization after 48-72 h. No complete or partial responses were observed. In one patient, a 40% regression of a tumor lesion was noted. In conclusion, rhIL 12 at these dose levels and schedule was well tolerated and resulted in measurable immunological responses. PMID- 12912942 TI - Phase I and pharmacokinetic study of brostallicin (PNU-166196), a new DNA minor groove binder, administered intravenously every 3 weeks to adult patients with metastatic cancer. AB - PURPOSE: Brostallicin (PNU-166196) is a cytotoxic agent that binds to the minor groove of DNA with significant antitumor activity in preclinical studies. This trial was designed to determine the maximum tolerated dose, the toxicity profile, and the pharmacokinetics of Brostallicin in cancer patients. EXPERIMENTAL DESIGN: Patients were treated with escalating doses of Brostallicin ranging from 0.85 to 15 mg/m(2) administered as a 10-min i.v. infusion every 3 weeks. Blood samples for pharmacokinetic analysis were collected during the first and second course, and analyzed by liquid-chromatography with tandem-mass spectrometric detection. RESULTS: Twenty-seven evaluable patients received a total of 73 courses. Grade 4 neutropenia was the only dose-limiting toxicity at 12.5 mg/m(2), whereas grade 4 thrombocytopenia (1 patient) and grade 4 neutropenia (2 patients) were the dose limiting toxicities at 15 mg/m(2). Other side effects, including thrombocytopenia and nausea, were generally mild. The maximum tolerated dose was defined at 10 mg/m(2). The clearance and terminal half-life of Brostallicin were dose independent, with mean (+/-SD) values of 9.33 +/- 2.38 liters/h/m(2) and 4.69 +/- 1.88 h, respectively. There was no significant accumulation of Brostallicin with repeated administration. Significant relationships were observed between systemic exposure to Brostallicin and neutrophil counts at nadir. One partial response was observed in a patient with a gastrointestinal stromal tumor. CONCLUSION: Brostallicin was found to be well tolerated, with neutropenia being the principal toxicity. The recommended dose for additional evaluation in this schedule is 10 mg/m(2). PMID- 12912943 TI - Phase I dose-escalation study of the safety and pharmacokinetics of atrasentan: an endothelin receptor antagonist for refractory prostate cancer. AB - PURPOSE: Evidence suggests that endothelin (ET)-1 and its primary receptor, the ET(A) receptor, may contribute to the progression of prostate and other cancers. Atrasentan (ABT-627) is a highly potent, selective ET(A) receptor antagonist. This study assessed safety, maximum tolerated dose, and pharmacokinetics (PK) in patients with refractory adenocarcinomas, primarily prostate cancer. EXPERIMENTAL DESIGN: This 28-day, single-center Phase I trial evaluated the safety and PK of escalating oral atrasentan doses (2.5-95 mg) given daily (except day 2) to eligible patients >/==" BORDER="0">18 years old with an adenocarcinoma proven resistant to standard therapy. Priority was given to patients with hormone refractory prostate cancer. After 28 days, patients without objective signs of tumor progression were eligible to continue atrasentan in an extension study. RESULTS: Thirty-nine patients (30 of whom had prostate cancer) were treated in cohorts of three patients each with escalating atrasentan doses (2.5, 5, 10, 20, 30, 45, 60, 75, and 95 mg). The most common adverse events were rhinitis, headache, and peripheral edema. Anemia consistent with a reversible hemodilution effect was observed. No maximum tolerated dose was found in the dose range studied. Atrasentan PK were characterized by rapid absorption (mean T(max) = 0.9 h), mean +/- SD oral clearance of 24 +/- 15 liters/h, and volume distribution of 726 +/- 477 liters. PK were approximately dose-proportional and time independent across doses. CONCLUSIONS: Atrasentan is well tolerated, with no dose-limiting adverse events observed up to 95 mg. Adverse events are consistent with the vasodilatory effect of the drug. PK are linear and dose-proportional; the half life is appropriate for once-daily dosing. PMID- 12912945 TI - The RASSF1A tumor suppressor gene is commonly inactivated in adenocarcinoma of the uterine cervix. AB - PURPOSE: Development of adenocarcinoma (AC) of the uterine cervix, as well as squamous cell carcinoma (SCC), is strongly linked to infection by high-risk human papillomavirus (HPV) types. Human HPV E6 and E7 proteins inactivate the tumor suppressor genes p53 and retinoblastoma, respectively. However, additional genetic alterations may be required to maintain a malignant phenotype. Allelic loss at the short arm of chromosome 3 is one of the most frequent genetic changes found in cervical cancer and various other types of human cancer, including lung, breast, and ovarian cancer. This implies that a resident tumor-suppressor gene in this region is involved in the genesis of these tumors. RASSF1A, which is located at 3p21.3, is rarely inactivated by mutations but has been suggested as a target tumor suppressor gene on the basis of its frequent inactivation through promoter hypermethylation and loss of heterozygosity in a variety of primary human cancers. In the present study, we sought to determine whether epigenetic silencing of RASSF1A caused by hypermethylation of the promoter region plays a role in the development of uterine cervical cancer. EXPERIMENTAL DESIGN: We studied 51 uterine cervical carcinoma samples. These 51 cases were comprised of 31 SCCs and 20 ACs. Real-time methylation-specific PCR system was used for the detection and quantitation of the bisulfite-converted methylated version of the RASSF1A promoter region. The 20 cases of cervical AC were also analyzed for the presence of oncogenic HPV 16 DNA using a PCR-based method. RESULTS: We found complete methylation of the RASSF1A promoter in 45% (9 of 20 samples) of AC cases. There was no promoter methylation observed in any of the 31 cases of SCC. We also correlated RASSF1A promoter hypermethylation to oncogenic HPV 16 infection. HPV 16 DNA was found in 3 of 9 (33%) AC tumors with RASSF1A promoter hypermethylation and 5 of 11 (45%) AC tumors without RASSF1A promoter hypermethylation. We could not demonstrate an inverse correlation between RASSF1A methylation and HPV 16 infection in AC of the uterine cervix. CONCLUSIONS: Hypermethylation of the RASSF1A promoter region is common in AC of the uterine cervix and rare in squamous carcinoma of uterine cervix. HPV infection does not correlate with RASSF1A methylation status in AC of the uterine cervix, but the absence of RASSF1A methylation in SCC of the uterine cervix coupled with the high incidence of HPV 16 infection in this subtype is in accord with previous reports. Our results suggest that epigenetic silencing of RASSF1A may play a role in the development of AC of the uterine cervix. PMID- 12912944 TI - Recombinant fowlpox viruses encoding the anchor-modified gp100 melanoma antigen can generate antitumor immune responses in patients with metastatic melanoma. AB - PURPOSE: The purpose of this study was to evaluate the immunological responses and therapeutic effectiveness of immunization with fowlpox vaccines encoding the gp100 melanoma antigen in patients with metastatic melanoma. EXPERIMENTAL DESIGN: In three consecutive clinical trials, patients were immunized with recombinant fowlpox viruses encoding three different forms of the melanoma/melanocyte associated antigen gp100: (a) the native, full-length gp100 molecule; (b) the gp100 molecule with two amino acids modified to increase binding to HLA-A*0201 molecules; and (c) a "minigene" construct encoding a single, modified epitope gp100:209-217(210M) targeted to the endoplasmic reticulum. The immunogenicity of these constructs was studied using peripheral blood mononuclear cells to measure epitope-specific release of IFN-gamma. RESULTS: Reactivity against gp100 was not seen in any patient before receiving fowlpox immunization. Whereas just one of seven patients developed reactivity after receiving fowlpox encoding native gp100, 10 of 14 patients who received fowlpox encoding the anchor modified full length gp100 exhibited reactivity against the native gp100 molecule, and 12 of 16 patients were successfully immunized after inoculation with the modified minigene construct (p2 = 0.02). There was no difference in the latter group between those randomized to vaccination by i.v. or i.m. routes. There was one partial cancer regression in the group of 46 patients receiving virus in the absence of interleukin (IL)-2. Once patients showed evidence of progressive disease, they were eligible for "cross-over" treatment to IL-2 alone or with the fowlpox virus. None of the 13 patients receiving the full-length or modified full-length forms of gp100 responded when receiving IL-2, whereas 6 of 12 patients who received the fowlpox containing the minigene construct and then received IL-2 showed objective cancer regressions, including three patients with complete regression. CONCLUSIONS: These data underscore the importance of modifying anchor residues of nonmutated self-antigen peptides to generate cellular immune responses after immunization and support the further investigation of recombinant fowlpox viruses encoding modified epitopes administered in combination with IL-2. PMID- 12912946 TI - DNA copy number changes in cervical adenocarcinoma. AB - PURPOSE: There is evidence that specific genetic events are involved in the initiation and progression of squamous cell carcinoma of the uterine cervix. The genotype-phenotype correlations in cervical adenocarcinoma (AC) are unclear. EXPERIMENTAL DESIGN: Comparative genomic hybridization was applied to screen for DNA copy number gains and losses in 22 cervical ACs of clinical stage IB. IHC was performed in all of the samples to determine HER-2/neu expression (HercepTest). RESULTS: The most frequent copy number alterations were DNA sequence gains of chromosome 17q (54%). HER-2/neu expression (score 2+) was immunohistochemically detected in 2 of 22 tumors. DNA sequence losses were most prevalent on chromosomes Xq (50%), Xp (36%), 18q (36%), and 4q (36%). DNA sequence losses of chromosome 18q were associated significantly with poor prognosis in cervical AC (P < 0.01). CONCLUSIONS: DNA sequence copy number gains of chromosome 17q are frequent events in ACs of the cervix. However, gains on 17q are not associated with HER-2/neu expression in cervical ACs. The inactivation of tumor suppressor genes on chromosome 18q might be responsible for the progression of both cervical AC and cervical squamous cell carcinoma. PMID- 12912947 TI - Prediction of response to docetaxel by quantitative analysis of class I and III beta-tubulin isotype mRNA expression in human breast cancers. AB - PURPOSE: The relationship of intratumoral mRNA levels of class I and III beta tubulin isotypes with clinical response to docetaxel has been studied in breast cancer patients. EXPERIMENTAL DESIGN: Expression of class I and class III beta tubulin mRNA levels was determined by a real-time PCR in breast cancer tissues obtained from 39 patients with locally advanced breast cancers (n = 26) or locally recurrent breast cancers (n = 13) before docetaxel treatment. RESULTS: Class I beta-tubulin mRNA levels of responders [6.58 +/- 1.43 (x10(2)), mean +/- SE] were significantly (P = 0.002) lower than those of nonresponders [14.97 +/- 2.95 (x10(2))], and class III beta-tubulin mRNA levels of responders (1.38 +/- 0.40) were also significantly (P = 0.003) lower than those of nonresponders (7.43 +/- 2.77). Breast cancers were divided into four groups according to the expression status of class I and class III beta-tubulin isotype mRNA, i.e., the class I-high/class III-high group (n = 13), the class I-high/class III-low group (n = 7), the class I-low/class III-high group (n = 7), and the class I-low/class III-low group (n = 12). The class I-high/class III-high group showed a very low response rate (15%), whereas the class I-low/class III-low group showed a very high response rate (75%). The class I-high/class III-low group and the class I low/class III-high group showed intermediate response rates of 57% and 43%, respectively. CONCLUSIONS: These results demonstrate that high expression of class I and class III beta-tubulin isotype mRNA is significantly associated with docetaxel resistance, and determination of class I and class III beta-tubulin isotype mRNA levels would be useful in the prediction of response to docetaxel. PMID- 12912948 TI - LKB1 protein expression in the evolution of glandular neoplasia of the lung. AB - PURPOSE: About one-third of sporadic lung adenocarcinomas demonstrates biallelic inactivation of the LKB1 gene, but the timing of this event is not known. DESIGN: We performed LKB1 immunohistochemistry on 35 primary lung adenocarcinomas and 96 atypical adenomatous hyperplasias (AAH), a form of early glandular neoplasia from which some lung adenocarcinomas arise. RESULTS: In all cases, strong cytoplasmic staining was noted in the non-neoplastic epithelium lining the airways from the bronchi to the terminal bronchioles. There was a marked reduction in LKB1 staining in 9 of 35 (26%) adenocarcinomas and in 10 of 96 (10%) AAHs. When the AAHs were subclassified on the basis of cytoarchitectural atypia, loss of LKB1 expression was more frequent in the high-grade lesions (7 of 33, 21%) than low grade lesions (3 of 63, 5%; P = 0.021). For the 21 adenocarcinomas where the genetic status was known, immunohistochemistry staining reliably reflected the activational state of the LKB1 gene (95% concordancy). CONCLUSIONS: In AAH, loss of LKB1 expression is strongly associated with severe dysplasia, suggesting that LKB1 inactivation may play a role in the critical transition from premalignant to malignant tumor growth. PMID- 12912949 TI - Sensitive and specific detection of circulating cancer cells in patients with hepatocellular carcinoma; detection of human telomerase reverse transcriptase messenger RNA after immunomagnetic separation. AB - PURPOSE: We evaluated whether detection of human telomerase reverse transcriptase (hTERT) mRNA after immunomagnetic separation is useful to detect circulating cancer (CC) cells. EXPERIMENTAL DESIGN: Two ml of peripheral blood were collected from 55 cases with hepatocellular carcinoma (HCC), 20 cases with chronic liver diseases devoid of cancer, and 20 healthy volunteers. Then 1500 and 500 micro l were subjected to immunomagnetic separations using Ber-EP4 and anti-CD45 antibodies, harvested and supernatant cells were collected as epithelial and nonleukocyte fractions, respectively. Samples of each fraction were subjected to reverse transcription-PCR detecting beta-actin, interleukin-2 receptor (IL-2r), alpha-fetoprotein, and hTERT mRNAs. The cases were judged to be positive, equivocal, or negative for CC cells when hTERT positivity with IL-2r negativity, hTERT positivity with IL-2r positivity, or hTERT negativity was seen in epithelial and/or nonleukocyte fractions, respectively. RESULTS: The dilution experiments revealed that our system could detect 10(0-1) HeLa cells involved in 2 ml of blood. The Ber-EP4-harvested cells from cases with distant metastasis were positive for immunostaining using Hep Par 1 monoclonal antibody. CC cells were judged to be positive in 29 of 55 (53%) HCC cases. On the contrary, no cases without HCC were determined to be positive. The frequency of positivity was significantly correlated with disease extent of HCC. CONCLUSIONS: These results strongly suggest that detection of hTERT mRNA after immunomagnetic separation is a specific and sensitive tool to detect CC cells and that it would provide useful source for further investigation of cancer metastasis. PMID- 12912950 TI - An intron splice acceptor polymorphism in hMSH2 and risk of leukemia after treatment with chemotherapeutic alkylating agents. AB - PURPOSE: We sought to determine whether the -6 exon 13 T>C polymorphism in the DNA mismatch repair gene hMSH2 modulates susceptibility to acute myeloid leukemia after therapy and particularly after O(6)-guanine alkylating chemotherapy. We also determined the extent of microsatellite instability (MSI) in therapy-related acute myeloid leukemia (t-AML) as a marker of dysfunctional DNA mismatch repair. EXPERIMENTAL DESIGN: Using a novel restriction fragment length polymorphism, verified by direct sequencing, we have genotyped 91 t-AML cases, 420 de novo acute myeloid leukemia cases, and 837 controls for the hMSH2 -6 exon 13 polymorphism. MSI was evaluated in presentation bone marrow from 34 cases using the mononucleotide microsatellite markers BAT16, BAT25, and BAT26. RESULTS: Distribution of the hMSH2 -6 exon 13 polymorphism was not significantly different between de novo acute myeloid leukemia cases and controls, with heterozygotes and homozygotes for the variant (C) allele representing 12.2 and 1.6%, respectively, of the control population. However, the variant (C) hMSH2 allele was significantly overrepresented in t-AML cases that had previously been treated with O(6)-guanine alkylating agents, including cyclophosphamide and procarbazine, compared with controls (odds ratio, 4.02; 95% confidence interval, 1.40-11.37). Thirteen of 34 (38%) t-AML cases were MSI positive, and 2 of these 13 cases were homozygous for the variant (C) allele, a frequency substantially higher than in the control population. CONCLUSIONS: Association of the hMSH2 -6 exon 13 variant (C) allele with leukemia after O(6)-guanine alkylating agents implicates this allele in conferring a nondisabling DNA mismatch repair defect with concomitant moderate alkylation tolerance, which predisposes to the development of t-AML via the induction of DNA mismatch repair-disabling mutations and high-grade MSI. Homozygosity for the hMSH2 variant in 2 of 13 MSI-positive t-AML cases provides some support for this model. PMID- 12912951 TI - Denaturing high performance liquid chromatography analysis of the DPYD gene in patients with lethal 5-fluorouracil toxicity. AB - Dihydropyrimidine dehydrogenase (DPD) enzyme deficiency is a pharmacogenetic syndrome with possible fatal outcome following 5-fluorouracil (5-FU) treatment. Several studies examining the molecular basis for DPD deficiency have identified over 30 sequence variations in the DPYD gene (which codes for the DPD enzyme). Our laboratory has recently developed and validated a denaturing high performance liquid chromatography method capable of identifying both known and unknown sequence variations in the DPYD gene. In the present study, we used this denaturing high performance liquid chromatography approach to examine the DPYD genotype of three patients who experienced lethal toxicity after administration of 5-FU. DPD enzyme activity could only be measured in one patient before death and demonstrated that lethal toxicity can occur in a partially DPD-deficient individual. Multiple heterozygous sequence variations (both known and unknown) were detected in all three patients including the novel variants 545T>A, M182K and 2329G>T, A777S. We conclude that (a) lethal toxicity can occur in partially DPD-deficient individuals after administration of 5-FU and is not exclusive to profoundly DPD-deficient individuals as suggested previously, (b) the complicated heterozygote genotype seen in these patients, combined with DPD deficiency being an autosomal codominant inherited syndrome, precludes the use of simple genotyping assays that identify only one or two mutations as a method for identifying DPD-deficient individuals; and (c) these multiple heterozygote genotypes (which are more difficult to accurately characterize) may be responsible for some of the conflicting reports which suggests a lack of correlation between phenotype and genotype. PMID- 12912952 TI - Microsatellite changes in nipple aspirate fluid and breast tissue from women with breast carcinoma or its precursors. AB - PURPOSE: Loss of heterozygosity (LOH) and microsatellite instability (MSI) have been identified in a variety of human cancers. The purpose of this prospective study was to determine whether (a) DNA can be isolated from nipple aspirate fluid (NAF) and PCR amplified to large fragments, (b) LOH and MSI are detectable in NAF, and (c) LOH and MSI in tissue and NAF increase with disease progression from precursor lesions to cancer. EXPERIMENTAL DESIGN: Forty-six matched samples from breast lesions, normal breast, and NAF were microdissected, and DNA was extracted. Eleven microsatellite markers from seven chromosomes that have a high frequency of LOH/MSI in breast cancer were designed and respectively amplified. RESULTS: LOH and/or MSI were identified in 22 of 46 (48%) breast lesions, including LOH in 8 of 36 (22%) proliferative/papilloma (P/Pap) and 7 of 10 (70%) cancer specimens, whereas MSI was found in 14 of 36 (39%) P/Pap and 6 of 10 (60%) cancer specimens. LOH/MSI loci in which alterations were detected in the 22 tissue specimens were PCR amplified using matched NAF DNA. LOH/MSI was detected in NAF from both P/Pap (5 of 15; 33%) and breast cancer (3 of 7; 43%) samples. CONCLUSIONS: Our findings suggest that (a) DNA from NAF, a physiological fluid collected noninvasively, can be PCR amplified and used to screen for LOH and MSI alterations that are known to be linked to breast cancer, suggesting that this methodology might prove useful for breast cancer screening, and (b) similar to findings in breast tissue, LOH and MSI alterations increase in frequency with disease progression in NAF, which suggests that NAF is a surrogate for breast tissue which has important prognostic implications. PMID- 12912953 TI - Epigenetic down-regulation of death-associated protein kinase in lung cancers. AB - PURPOSE: Death-associated protein kinase (DAPK) is a pro-apoptotic serine/threonine kinase involved in apoptosis. Aberrant methylation of DAPK was reported in lung cancers by methylation-specific PCR. However, we were unable to relate methylation with gene silencing with the same methodology. Our goals were to develop a methodology that related methylation with gene silencing and use it to study the state of the gene in lung cancers. EXPERIMENTAL DESIGN AND RESULTS: Using a semiquantitative real-time reverse transcription-PCR, DAPK expression was lower in lung cancers than in corresponding nonmalignant bronchial epithelial cells in five of six primary short-term cultures. In continuous cell lines, mRNA expression was down-regulated, as well as compared with nonmalignant bronchial epithelial cells, and its protein was not detected by Western blotting in 17 of 23 (74%) cell lines. We investigated methylation status of 5' flanking region of DAPK by combined bisulfite restriction analysis and bisulfited DNA sequencing. Aberrant methylation was detected in 21 of 48 (44%) cell lines, 2 of 6 primary cultured tumors, and 14 of 38 (37%) primary lung cancers, although varying degrees of methylation were noticed. Furthermore, bisufite sequence data suggested that aberrant methylation might occur selectively at some CpG dinucleotides in cell lines which had absent expression. Treatment with 5-aza-2' deoxycytidine restored DAPK expression in heavily methylated cell lines tested, and histone deacetylase inhibitor trichostatin A alone restored DAPK expression in some methylated cell lines as well. CONCLUSIONS: Our major findings are: (a) DAPK expression is frequently down-regulated in lung cancers; (b) aberrant methylation of DAPK is frequent in lung cancers, although considerable heterogeneity of methylation is present, and some specific CpG dinucleotides are often methylated in expression negative lung cancers; and (c) besides methylation and histone deacetylation, there may be other mechanisms for down-regulation of DAPK expression. PMID- 12912954 TI - Promoter hypermethylation of high-in-normal 1 gene in primary nasopharyngeal carcinoma. AB - PURPOSE: The methylation of high-in-normal-1 (HIN-1) gene promoter in undifferentiated nasopharyngeal carcinoma (NPC) is studied. EXPERIMENTAL DESIGN: The methylation status of HIN-1 in NPC cell lines, primary NPC, paired nasopharyngeal swabs, paired throat-rinsing fluid, and paired peripheral blood was assessed by methylation-specific PCR assay. The relationship between HIN-1 promoter methylation and transcription in NPC cell lines was evaluated by reverse transcription-PCR and demethylation agent treatment (5-aza-2-deoxycytidine). RESULTS: Hypermethylated promoter was observed in five of five (100%) NPC cell lines and not found in three normal nasopharyngeal outgrowths, two tonsil epithelial cell cultures, and two skin fibroblast cultures. Reverse transcription PCR assay indicated that HIN-1 transcription was significantly down-regulated in the NPC cell line with promoter methylation. Treatment with demethylation agent, 5-aza-2-deoxycytidine, restored HIN-1 transcription in the NPC cell line. Methylated HIN-1 promoter was found in 36 of 47 (77%) primary NPC tumors and not found in the normal nasopharyngeal biopsies. Methylated HIN-1 promoter was detected in 12 of 26 (46%) nasopharyngeal swabs, 5 of 26 (19%) throat-rinsing fluids, 2 of 11 (18%) plasmas, and 5 of 11 (46%) buffy coats of peripheral blood of the NPC patients but was not detectable in all normal controls. CONCLUSION: HIN-1 promoter hypermethylation is common in NPC. Methylated promoter DNA in nasopharyngeal swab, throat-rinsing fluid, and peripheral blood might be potentially useful as tumor marker for screening of NPC. PMID- 12912955 TI - Serum levels of carboxy-terminal telopeptide of type-I collagen are elevated in patients with multiple myeloma showing skeletal manifestations in magnetic resonance imaging but lacking lytic bone lesions in conventional radiography. AB - PURPOSE: Skeletal involvement is a hallmark of multiple myeloma. Increased bone resorption can even be present in patients lacking osteolyses in conventional radiography. Magnetic resonance imaging (MRI) of the spine was established as a more sensitive technique to depict bone abnormalities. Type-I collagen degradation product carboxyterminal telopeptide of type-I collagen (ICTP) was introduced as a novel biochemical parameter reflecting the bone resorption activity in myeloma. The aim of this study was to evaluate whether increased ICTP serum levels predict abnormal MRI patterns in myeloma patients. EXPERIMENTAL DESIGN: MRI of the spine was performed in 32 untreated patients with multiple myeloma, who had no skeletal abnormalities in conventional radiographies. Simultaneously, ICTP was measured in serum by a competitive radioimmunoassay at corresponding time points. RESULTS: Serum ICTP was significantly (P = 0.002) elevated in patients with abnormal bone MRI compared with those patients with normal MRI findings. The sensitivity of ICTP for depiction of MRI abnormalities was 79%; the positive and negative predictive values were 85 and 84%, respectively. Compared with ICTP, the parameters of disease activity, beta2 microglobulin and C-reactive protein, had a much lower sensitivity for abnormal MRI (29 and 64%, respectively). CONCLUSIONS: In myeloma patients without osteolytic lesions in conventional radiography, abnormal skeletal MRI is accompanied by an increase in serum levels of ICTP. Our data show that ICTP can be used as an inexpensive parameter to identify myeloma patients with normal skeletal survey who have a high probability of skeletal involvement and deserve more accurate diagnostic evaluation using MRI. PMID- 12912956 TI - Expression and functional analyses of breast cancer resistance protein in lung cancer. AB - PURPOSE: Breast cancer resistance protein (BCRP/ABCG2), an ATP binding cassette half-transporter, confers resistance to mitoxantrone, doxorubicin, and topoisomerase I inhibitors of irinotecan and topotecan. Recently, we reported that BCRP efficiently transported SN-38 (the active metabolite of irinotecan) with a high affinity in lung cancer cells in vitro (K. Nakatomi et al., Biochem. Biophys. Res. Commun., 288: 827-832, 2001). The aim of this study is to explore the role of BCRP in the drug resistance of lung cancer. EXPERIMENTAL DESIGN: The BCRP mRNA expression in lung cancer cells and 23 untreated non-small cell lung cancer (NSCLC) tissues was quantitated by real-time reverse transcription-PCR. To evaluate the drug-efflux function of BCRP, the intracellular topotecan accumulation and drug sensitivity were measured in lung cancer cells with various levels of the BCRP mRNA expression by flow cytometric and tetrazolium dye assay, respectively. RESULTS: The levels of BCRP mRNA expression in the cell lines were significantly correlated with the BCRP function and the sensitivity to SN-38 and topotecan. In NSCLC tissues, the BCRP mRNA expression levels were widely dispersed. Five (22%) of 23 tissues expressed higher levels of the BCRP mRNA than that in NCI-H441 cells with active BCRP function conferring high resistance to topotecan in vitro. CONCLUSIONS: Some NSCLC tissues expressed sufficient levels of the BCRP mRNA to confer drug resistance in vitro. PMID- 12912957 TI - A transcriptional progression model for head and neck cancer. AB - PURPOSE: A genetic progression model for head and neck squamous cell carcinoma (HNSC) has been established and implies the presence of transcriptional dysregulation as a consequence of accumulation of genetic alterations. Although expression array data have been provided for HNSC, the timing of transcriptional dysregulation in the progression from normal mucosa to dyplastic epithelium to invasive HNSC has not been described. Here, we describe a transcriptional progression model of HNSC. EXPERIMENTAL DESIGN: Expression arrays representing >12,000 genes and expressed sequence tags were used to examine malignant lesions (M), premalignant lesions (PM), distant, histopathologically normal mucosa from patients with premalignant or malignant lesions (MN), and normal mucosa from the upper aerodigestive tract of patients with noncancer diagnoses (N). Significance analysis of microarrays, hierarchical clustering, and principal components analysis was used to identify genes with differential expression patterns. RESULTS: Using a false discovery rate of <5% for significance analysis of microarray, the M group revealed 965 up-regulated and 1106 down-regulated genes relative to the N group. The PM group demonstrated 108 up-regulated and 226 down regulated genes relative to the N group, whereas the M group demonstrated only 5 up-regulated and 13 down-regulated genes relative to the PM group. Both hierarchical cluster analysis and principal components analysis revealed a consistent separation between the N, PM, and M groups, with a closer association between the PM and M groups. To provide independent validation of the microarray data, quantitative reverse transcription-PCR was performed for a significantly up regulated gene, integrin alpha 6, correlating well with microarray data (linear regression analysis, P < 0.0001). CONCLUSIONS: Similarly to the genetic progression model of HNSC, this transcriptional model shows that the majority of alterations occurs before the development of malignancy and identifies key targets of transcriptional dysregulation during progression from a normal to a premalignant state and from a premalignant to a malignant state. PMID- 12912958 TI - Cyclooxygenase 2 expression and molecular alterations in Peutz-Jeghers hamartomas and carcinomas. AB - PURPOSE: Peutz-Jeghers syndrome (PJS) is a hamartomatous polyposis disorder with a high cancer risk. Debate exists about the premalignant potential of hamartomas. Also, treatment options other than surveillance are not available. Therefore, molecular alterations in hamartomas and PJS carcinomas were studied. The objective was (a) to evaluate expression of cyclooxygenase (COX)-2 as target for chemopreventive treatment and (b) to define the neoplastic potential of hamartomas at the molecular level. EXPERIMENTAL DESIGN: Paraffin-embedded samples of 24 PJS hamartomas, including 2 hamartomas with dysplastic changes, and 11 PJS carcinomas were available. Slides were stained with antibodies against COX-2, beta-catenin, cyclin D1, p21(waf1/cip1), Ki-67, and p53. DNA was studied for loss of heterozygosity (LOH) at 19p (STK11), 5q (APC), and 17p (TP53); mutations in beta-catenin, APC, and K-RAS; and microsatellite instability. RESULTS: Moderate or strong epithelial COX-2 was present in 25% of hamartomas, including two hamartomas with dysplastic changes, and 64% of carcinomas. Several hamartomas showed focal nuclear beta-catenin (18%) and cyclin D1 overexpression (29%), both unrelated to dysplasia at histological examination. Disturbed topographical expression of Ki-67 in relation to p21(waf1/cip1) was focally present in 27% of hamartomas, including those with dysplastic changes. Most carcinomas showed nuclear beta-catenin (71%), cyclin D1 overexpression (71%), and aberrant Ki-67 staining (100%). There was LOH at 19p in 32% of hamartomas and 82% of carcinomas. p53 staining was present in four (36%) carcinomas, one of which showed LOH at 17p. No beta-catenin mutations were found. APC mutations were present in two carcinomas, but LOH at 5q was not found. Two carcinomas had K-RAS mutations, and one carcinoma had microsatellite instability. CONCLUSIONS: The presence of COX-2 expression in PJS carcinomas and dysplastic hamartomas provides a rationale for chemoprevention with nonsteroidal anti-inflammatory drugs or COX-2 inhibitors. Focal immunohistochemical changes, which may indicate a premalignant potential, were present in some nondysplastic PJS hamartomas. Molecular changes in carcinomas and dysplastic hamartomas in PJS are distinct from the usual adenoma carcinoma sequence. PMID- 12912959 TI - beta-Catenin and ras oncogenes detect most human colorectal cancer. AB - PURPOSE AND STUDY DESIGN: Recent studies have shown that beta-catenin translocated into the cell nucleus functions like an oncogene. Accumulating evidence suggests that activation of the beta-catenin oncogenic signaling cascade along with its twin, the K-ras cascade, may exert syngeneic or synergistic effects on tumor development and progression. In the study reported here, we analyzed oncogenic beta-catenin activation on the basis of its nuclear accumulation (NA) and compared the results with those of mutational activation of K-ras in 74 patients with colorectal cancer to determine whether the two oncogene mediated signaling cascades interact. RESULTS: We found two distinct patterns of beta-catenin activation, i.e., diffuse NA in 20 cases (27%) and selective NA at the tumor invasion front (NAinv) in 19 cases (26%). The presence of the NAinv pattern was significantly correlated with advanced Dukes' stage tumor (P = 0.0005) and the presence of distant metastases (P = 0.0064). K-ras proto-oncogene was mutated in the tumors of 31 cases (42%). Activated beta-catenin or K-ras was detected in most (78%) colorectal cancers analyzed, although a weak inverse correlation was found between the activities of the two oncogenes in the tumors. Importantly, most (7 of 8) patients with tumor showing both K-ras activation and the NAinv pattern of beta-catenin activation were in Dukes' stage C at surgery, and half of them developed distant metastases to the liver and lungs. CONCLUSION: The results suggest that although oncogenic activation of beta-catenin and K-ras is independent in the process of clinical cancer development, combined analysis of the two major oncogenes can detect most colorectal cancers and identify a subset of patients with poorer outcomes. Consequently, activation of either or both of these oncogenes may serve as a genetic marker for molecular diagnosis. PMID- 12912960 TI - Gene expression profiling of malignant mesothelioma. AB - PURPOSE: Malignant mesothelioma is a uniformly fatal cancer of the pleural and peritoneal spaces. Several challenging clinical problems include poor understanding of the pathophysiology, inaccurate diagnosis from tissue samples, and unsuccessful treatment strategies. The purpose of this study was to use microarray analysis to identify specific gene expression changes in mesothelioma compared with normal mesothelium. EXPERIMENTAL DESIGN: We performed gene expression analysis on mesothelioma tissue specimens from 16 patients and compared these to 4 control pleural tissue samples using cDNA microarray filters with 4132 clones. Multiple normalization and analysis approaches were used. Quantitative reverse transcription-PCR and immunohistochemistry were used to validate results. RESULTS: Genes (166) were significantly up-regulated, and 26 were down-regulated. Validation of 18 genes using real-time PCR confirmed array predictions in every case. Analysis revealed activation of several key pathways including genes involved in glucose metabolism, mRNA translation, and cytoskeletal remodeling. Expression profiling identified processes likely responsible for 18-fluoro-2-deoxy-glucose uptake and tumor localization by positron emission tomography, and a role for hypoxia-inducible factor-1 was suggested. Potentially important up-regulated genes included gp96, lung resistance-related protein, galectin-3 binding protein, the M(r) 67,000 laminin receptor (on tumor vessels), and voltage-dependent anion channels. Prospective testing using reverse transcription-PCR confirmed up-regulation of these novel markers. CONCLUSIONS: Expression profiling revealed marked up-regulation of energy, protein translation, and cytoskeletal remodeling pathways in mesothelioma. Additional genes that could be important in our understanding of the pathogenesis of mesothelioma, aiding in diagnosis, or improving targets for therapy were also identified. PMID- 12912961 TI - Morphometric evaluation of tumor matrix metalloproteinase 9 predicts survival after surgical resection of adenocarcinoma of the lung. AB - PURPOSE: Recently, several matrix metalloproteinases (MMPs) have shown promise as prognosticators in non-small cell lung cancer. In this study, we sought to validate the importance of MMP-9 and to study the relationships between MMP-9 and several other tumor or stromal markers. EXPERIMENTAL DESIGN: We examined MMP-9 and several other markers in tumor tissues from 152 patients with surgically excised adenocarcinomas of the lung. Their preoperative clinical stages were T(1 4)N(0)M(0); however, pathological exam of their resected tissues demonstrated that 33 were stage II, and 64 were stage III. We used immunohistochemistry and morphometry to evaluate the amount of tumor staining for MMP-9, and the outcome for our study was survival time until death from recurrent lung cancer. RESULTS: Multivariate Cox model analysis demonstrated that pathological stage was significantly related to survival time (P < 0.01), but quantitative staining of the tumor for MMP-9 added prognostic information (P < 3.0 x10(-16)) and was more strongly prognostic than pathological stage. In the subset of pathological stage I patients, staining for MMP-9 was also significantly associated with survival (P < 1.0 x10(-6)), and a cutpoint at the median staining of 11.2% for MMP-9 divided them into two groups with distinctive survival times. Those with MMP-9 > 11.2% had a median survival time of just 11 months. Those with MMP-9 < 11.2% had not reached a median survival and had a mean survival time of >62 months. CONCLUSIONS: Tumor staining for MMP-9 in resected adenocarcinoma of the lung is strongly related to survival. Patients with >11.2% staining in their tumors comprise a subset with a high hazard for dying of lung cancer and may be an appropriate target for prospective studies of adjuvant chemotherapy after surgical resection. PMID- 12912962 TI - CpG oligodeoxynucleotides potentiate the antitumor effects of chemotherapy or tumor resection in an orthotopic murine model of rhabdomyosarcoma. AB - PURPOSE: CpG oligodeoxynucleotides (ODNs) are synthetic DNA sequences that mimic bacterial DNA and have potent immunostimulatory effects on dendritic cells (DCs), B cells, and natural killer cells. To evaluate CpG ODN antitumor effects against solid tumors, we used an orthotopic murine model of embryonal rhabdomyosarcoma. EXPERIMENTAL DESIGN: The systemic administration of CpG 2006 was tested beginning on day 9 or 19 when tumors were not yet palpable or palpable, respectively, and after surgical resection of the tumor. CpG was also administered in combination with the chemotherapeutic agents, cyclophosphamide (CY) and topotecan, or surgical resection. RESULTS: Systemic CpG prolonged survival when begun at day 9 but had no effect with a large tumor burden. CpG administered after surgical resection of tumor significantly improved survival of mice (P < 0.03). On day 9, CY plus CpG 2006 resulted in improved survival compared with CY alone (70% versus 41%, respectively). Survival was significantly improved when CpG 2006 was administered systemically with CY beginning on day 19 (15% versus 0% survival). The administration of CpG 2006 with topotecan significantly improved survival in mice with large tumors. Cell-depletion studies demonstrated that the antitumor effects of systemically administered CpG 2006 combined with CY were predominantly T cell dependent. CONCLUSIONS: These data are the first to show that immune stimulatory agents such as CpGs may enhance the antitumor effects of chemotherapeutic agents and improve survival after surgical resection of a solid tumor. PMID- 12912963 TI - Tannic acid is an inhibitor of CXCL12 (SDF-1alpha)/CXCR4 with antiangiogenic activity. AB - PURPOSE: Increasing evidence suggests that interaction between the chemoattractant CXCL12/stromal cell-derived factor-1alpha and its receptor CXCR4 plays a pivotal role in the metastasis of various tumors. Our previous studies showed that multi-component Chinese herbal medicines inhibited the effects of CXCL12/CXCR4. As a result of sequential chromatographic fractionation of one herbal medicine ingredient, Lianqiao (fruit of Forsythia suspensa), we observed that tannins were, at least in part, responsible for this activity. The aim of this study was to assess the anti-CXCL12/CXCR4 activity of a commercial tannic acid and evaluate its potential to inhibit tumor cell migration and angiogenesis in vitro. EXPERIMENTAL DESIGN: The inhibitory effect of tannic acid on CXCL12/CXCR4 was measured by chemotaxis assay, ligand binding assay, and fluorescence-activated cell sorter analysis. The antiangiogenic effect of tannic acid was assessed by in vitro endothelial cell tube formation. RESULTS: Tannic acid, at nontoxic concentrations, specifically inhibited CXCL12-induced human monocyte migration (IC(50), 7.5 micro g/ml) but did not inhibit CCL2-, CCL3-, CCL5-, formylmethionylleucylphenylalanine (fMLP)-, or C5a-induced migration. The compound markedly blocked CXCL12 binding to THP-1 cells (IC(50), 0.36 micro g/ml). Tannic acid also inhibited CXCL12-induced, but not epidermal growth factor induced, migration of MDA 231 breast tumor cells. Additionally, 0.5 micro g/ml of tannic acid selectively inhibited CXCL12-mediated, but not basic fibroblast growth factor- or endothelial cell growth supplement-mediated, bovine aorta endothelial cell capillary tube formation. CONCLUSION: These studies indicate that tannic acid is a novel selective CXCL12/CXCR4 antagonist and consequently may provide a mechanistic basis for the reported antitumor and anti-inflammatory properties of tannic acid. PMID- 12912964 TI - Interleukin 12-based immunotherapy improves the antitumor effectiveness of a low dose 5-Aza-2'-deoxycitidine treatment in L1210 leukemia and B16F10 melanoma models in mice. AB - PURPOSE: Recent findings indicating that many genes related to cancer development are silenced by an aberrant DNA methylation suggest that inhibitors of this process may be effective cancer therapeutics. In this study we investigated the efficacy of low-dose 5-aza-2'-deoxycitydine (DAC), a methylation inhibitor, with interleukin (IL) 12, one of the most potent cytokines with antitumor activity. EXPERIMENTAL DESIGN: Mice inoculated with L1210 leukemia cells or with B16F10 melanoma cells were treated with 7 daily injections of low-dose DAC (0.2 mg/kg) and/or 7 daily doses of IL-12 (100 ng/dose). Scid/scid mice as well as monoclonal antibodies against CD4, CD8, and NK1.1 were used to investigate the mechanisms of the antitumor effects of the combination treatment. The activity of murine lymphocytes was measured with enzyme-linked immunospot and (51)Cr release assays. RESULTS: Treatment with DAC or IL-12 given alone produced moderate antitumor effects. In both tumor models combined treatment resulted in potentiated antitumor effects and produced 70% long-term survivors among mice inoculated with L1210 cells. The antitumor efficacy of combined treatment was abrogated in scid/scid mice, and after depletion of CD4(+) and CD8(+) T cells. Mice inoculated with B16F10 melanoma cells had significantly delayed tumor growth after combined treatment with DAC and IL-12. Strong antitumor effect correlated with a significant activation of lymph node-derived CD8(+) and CD4(+) cells. Transient neutropenia was observed in mice under treatment of DAC alone, but remarkably this effect was not potentiated by IL-12. CONCLUSIONS: This study provides the first evidence that antitumor effects of DAC can be strongly potentiated by IL-12 and could be beneficial in an effective low-dose-based antitumor therapy. PMID- 12912965 TI - Role of antiapoptotic proteins in tumor necrosis factor-related apoptosis inducing ligand and cisplatin-augmented apoptosis. AB - PURPOSE AND EXPERIMENTAL DESIGN: The purpose of this study was to examine the effect of combined treatment with tumor necrosis factor-related apoptosis inducing ligand (TRAIL) and cisplatin in human head and neck squamous cell carcinoma. HNSCC-6 cells were treated with 0.1-1 micro g/ml TRAIL and/or 1-10 micro g/ml cisplatin for 24 h. RESULTS: TRAIL alone or cisplatin alone caused minimal cytotoxicity. The combination of TRAIL and cisplatin synergistically enhanced apoptotic death, caspase-8 and caspase-3 activation, as well as poly(ADP ribose) polymerase cleavage. However, the total cellular levels and the surface expression of TRAIL receptor proteins, such as death receptors 4 and 5 and decoy receptors 2 and 1, were not significantly changed by treatment with TRAIL and cisplatin. Interestingly, the level of the short form of Fas-associated death domain-like interleukin-1beta-converting enzyme-inhibitory protein (FLIP(S)) but not the long form of Fas-associated death domain-like interleukin-1beta converting enzyme-inhibitory protein was reduced through cleavage. Benzyloxycarbonyl-Asp-Glu-Val-Asp-fluoromethylketone a caspase-3 inhibitor, blocked the cleavage of FLIP(S) and caspase-3 activation. Overexpression of FLIP(S) protected cells from apoptotic death and FLIP(S) cleavage during treatment with TRAIL in combination with cisplatin. CONCLUSIONS: These results suggest that caspase-3 is responsible for FLIP(S) cleavage, and the cleavage of FLIP(S) is one of facilitating factors for TRAIL-induced apoptotic death. PMID- 12912966 TI - Protection of mice against Philadelphia chromosome-positive acute lymphoblastic leukemia by cell-based vaccination using nonviral, minimalistic expression vectors and immunomodulatory oligonucleotides. AB - PURPOSE: Childhood Philadelphia chromosome positive (Ph(+)) acute lymphoblastic leukemia (ALL) has a poor prognosis. Because leukemia cell burden is reduced but not eradicated by polychemotherapy, improved treatment strategies should enhance those immune mechanisms responsible for the maintenance of complete remission. The aim of this study was to evaluate the protection of mice challenged with the syngeneic Ph(+) ALL cell line BM185 using genetically modified leukemia cell vaccines and immunomodulating oligonucleotides. EXPERIMENTAL DESIGN: Because retroviral vectors are ineffective at transducing nondividing primary cells from human hematopoietic malignancies, we first evaluated nonviral techniques (electroporation and ballistic transfer) using minimalistic immunogenically defined gene expression vectors to generate B7.1 or granulocyte macrophage colony stimulating factor (GM-CSF)-expressing BM185 cells. Subsequently, protective vaccination experiments with these cells were performed in a leukemia challenge mouse model. RESULTS: Electroporation yielded a high transfection rate (82.6% for B7.1) with moderate GM-CSF secretion/1 x 10(6) cells (228 pg), whereas ballistic transfer led to a lower transfection rate (30.9%) with high GM-CSF secretion (614 pg). Secondly, we immunized mice with B7.1/interleukin 2- or B7.1/GM-CSF expressing BM185 cell vaccines. We observed a better protection of mice that received the B7.1/GM-CSF vaccine compared with these receiving the B7.1/interleukin 2 vaccine. Protection was additionally enhanced by application of a double stem-loop immunomodulating oligonucleotide containing CpG motifs. CONCLUSION: Our data indicate that immunization with B7.1/GM-CSF-expressing cell vaccines generated by electroporation and application of double stem-loop immunomodulating oligonucleotide protected mice against a murine Ph(+) ALL challenge. Ultimately, this approach may also lead to clinical benefit in patients with Ph(+) ALL. PMID- 12912967 TI - Ibuprofen-mediated reduction of hypoxia-inducible factors HIF-1alpha and HIF 2alpha in prostate cancer cells. AB - PURPOSE: Hypoxia-inducible factors HIF-1alpha and HIF-2alpha are considered to be potential targets for antineoplastic therapy because they regulate the expression of genes that contribute to tumor cell survival, aggressiveness, and angiogenesis. Nonsteroidal anti-inflammatory drugs (NSAIDs) have gained considerable interest as anticancer agents because of their cytotoxic and antiangiogenic properties. The aim of this study was to investigate whether NSAIDs inhibit HIFs and HIF-regulated gene expression in prostate cancer cells. EXPERIMENTAL DESIGN: PC3 and DU-145 cells were treated with ibuprofen (Ibu) and other NSAIDs under normoxic and hypoxic (95% N(2), 5% CO(2); <10 ppm O(2)) conditions. The effect of NSAIDs on HIF proteins was analyzed by Western blot analysis. HIF-regulated proteins, vascular endothelial growth factor (VEGF) and glucose transporter-1 (Glut-1), were analyzed by ELISA and Western blot analysis, respectively. RESULTS: Exposure of PC3 and DU-145 cells to hypoxic condition up regulated HIF-1alpha and HIF-2alpha proteins. Treatment with Ibu under normoxic and hypoxic conditions reduced the level of HIF-1alpha and HIF-2alpha. Ibu mediated down-regulation of HIFs was associated with down-regulation of HIF regulated proteins VEGF and Glut-1 in cells exposed to hypoxia. Other nonspecific NSAIDs, diclofenac and ketorolac, also inhibited HIF-1alpha and HIF-2alpha. The reduction in HIFs was observed in PC3 cells that expressed cyclooxygenase-2 (COX 2) protein as well as in DU-145 cells, which did not express COX-2 protein. COX-2 specific inhibitor NS-398 did not inhibit HIF-1alpha or VEGF and GLUT-1. CONCLUSIONS: These data indicate that one of the effects of NSAIDs is to reduce HIF protein levels. The inhibition of HIFs by NSAIDs was COX-2 independent. PMID- 12912968 TI - Cell-based protein delivery system for the inhibition of the growth of pancreatic cancer: NK4 gene-transduced oral mucosal epithelial cell sheet. AB - PURPOSE: Pancreatic resection for pancreatic cancer is the only curative modality, but the high incidence of local recurrence after surgery results in a very poor prognosis. This study aims to develop a new therapeutic tool that could inhibit the growth of remnant cancer cells, which is based on local delivery of NK4 (hepatocyte growth factor antagonist) secreted from an NK4 gene-transduced oral mucosal epithelial cell (OMEC) sheet (NK4-sheet), which is adhered to the resected surface. EXPERIMENTAL DESIGN: OMECs, harvested and cultured according to 3T3 feeder layer technique, were seeded on a collagen mesh-overlayered, biodegradable VICRYL mesh to produce an OMEC sheet. NK4 gene transduction was mediated by recombinant adenovirus (Ad-NK4). Applicability of OMECs for cell based NK4 delivery was examined. An experimental model using nude mice was established to determine the effect of an NK4-sheet on both tumor growth and angiogenesis. RESULTS: NK4 secreted from Ad-NK4-transduced OMECs suppressed MRC-5 induced invasion of pancreatic cancer cell lines. Heterotopically implanted gene transduced OMECs remained for >/==" BORDER="0">10 days while gradually decreasing. NK4-sheets inhibited both angiogenesis and tumor growth in vivo. CONCLUSION: Autologous OMEC was found to be suited to this purpose because of no secretion of hepatocyte growth factor, ease in harvesting from a patient, reasonably high proliferation potential, and no immune reaction. Although NK4 sheets under development exhibited a low level and short period of NK4 secretion, it is expected that this system may have a great potentiality of protein delivery system to target tissue at clinical situations when it is loaded with multilayered OMECs. PMID- 12912969 TI - Fully human anti-interleukin 8 antibody inhibits tumor growth in orthotopic bladder cancer xenografts via down-regulation of matrix metalloproteases and nuclear factor-kappaB. AB - PURPOSE: We previously demonstrated that overexpression of interleukin 8 (IL-8) in human transitional cell carcinoma (TCC) resulted in increased tumorigenicity and metastasis. This increase in tumor growth and metastasis can be attributed to the up-regulation in the expression and activity of the metalloproteinases MMP-2 and MMP-9. EXPERIMENTAL DESIGN: To investigate whether targeting IL-8 with a fully human anti-IL-8 antibody (ABX-IL8) could be a potential therapeutic strategy for controlling TCC growth, we studied its effects on TCC growth in vitro and in an in vivo mouse model. Human TCC cell lines 253J B-V and UM UC3 (high IL-8 producers), 253J (low IL-8), and 253J transfected with the IL-8 gene (high producer) were used. RESULTS: ABX-IL8 had no effect on TCC cell proliferation in vitro. However, in the orthotopic nude mouse model, after 4 weeks of treatment (100 micro g/week, i.p.), a significant decrease in tumor growth of both cell lines was observed. IL-8 blockade by ABX-IL8 significantly inhibited the expression, activity, and transcription of MMP-2 and MMP-9, resulting in decreased invasion through reconstituted basement membrane in vitro. The down-regulation of MMP-2 and MMP-9 in these cells could be explained by the modulation of nuclear factor-kappaB expression and transcriptional activity by ABX-IL8. CONCLUSIONS: Our data point to the potential use of ABX-IL8 as a modality to treat bladder cancer and other solid tumors, either alone or in combination with conventional chemotherapy or other antitumor agents. PMID- 12912970 TI - Activation of prodeath Bcl-2 family proteins and mitochondrial apoptosis pathway by sanguinarine in immortalized human HaCaT keratinocytes. AB - Sanguinarine, derived from the root of Sanguinaria canadensis and other poppy fumaria species, possesses strong antimicrobial, anti-inflammatory, and antioxidant properties. We earlier showed that sanguinarine kills human epidermoid carcinoma A431 cells via an induction of apoptosis [N. Ahmad et al., Clin. Cancer Res., 6: 1524-1528, 2000]. In this study, using immortalized human keratinocytes (HaCaT cells), we provide information about mechanism of the antiproliferative effect of sanguinarine. Sanguinarine [0.1 (M-2 (M)] treatment to HaCaT cells was found to inhibit in a dose-dependent manner the cell proliferation and induce apoptosis, as measured by 3-(4,5-dimethylthiazol-2-yl) 2,5-diphenyltetrazolium bromide assay and ELISA, respectively. Sanguinarine treatment also resulted in a significant cleavage of poly(ADP-ribose) polymerase in HaCaT cells. Because mitochondrial pathway is critical for the regulation of apoptosis, we studied the involvement and regulation of mitochondrial events in sanguinarine-mediated apoptosis of HaCaT cells. As shown by the immunoblot analysis, our data clearly demonstrated that sanguinarine treatment to HaCaT cells resulted in a dose-dependent (a) increase in the level of Bax with a concomitant decrease in Bcl-2 levels and (b) increase in Bax/Bcl-2 ratio. Sanguinarine also resulted in significant increases in the proapoptotic members of Bcl-2 family proteins, i.e., Bak and Bid. This was accompanied by increase in (a) protein expression of cytochrome c and apoptotic protease-activating factor-1 and (b) activity and protein expression of caspase-3, caspase-7, caspase-8, and caspase-9. Taken together, our data showed the involvement of mitochondrial pathway and Bcl-2 family proteins during sanguinarine-mediated apoptosis of immortalized keratinocytes. We suggest that sanguinarine could be developed as a drug for the management of hyperproliferative skin disorders, including skin cancer. PMID- 12912972 TI - Tumor growth retardation, cure, and induction of antitumor immunity in B16 melanoma-bearing mice by low electric field-enhanced chemotherapy. AB - PURPOSE: The exposure of cells in vitro to trains of low voltage-pulsed electric fields in the range of 20-100 V/cm was previously shown to induce an efficient uptake of macromolecules with molecular weight in the range of M(r) 300-2,000,000 via an endocytic-like process. This study examines the antitumor effectiveness of treatment based on similar exposure of solid tumors in mice to low electric fields (LEFs) in the presence of chemotherapeutic agents. EXPERIMENTAL DESIGN: LEF was applied to approximately 5 mm in diameter (60-70 mm(3)) s.c. B16-F10.9 melanoma tumors by percutaneously placed electrodes after intratumoral injection of either cis-platinum(II) diamminedichloride, Taxol, 5-fluorouracil, or bleomycin. RESULTS: Significant eradication of primary tumors, prolongation of survival, and complete cure of some of the C57Bl/6 mice from both primary tumors and metastases were achieved using this technique with cis-platinum(II) diamminedichloride, bleomycin, and Taxol (13.5, 8, and 26% cure rate, respectively). Mice cured by LEF-enhanced chemotherapy and challenged with a tumorigenic dose of B16-F10.9 cells lived significantly longer than first time inoculated ones, and 23.5% of the challenged mice did not develop tumors at all. Spleen cells from the cured mice that were inoculated together with B16-F10.9 cells inhibited the primary tumor growth in intact mice. Histological analysis of tumor sections of LEF-enhanced chemotherapy-treated mice revealed multiple necrotic areas, apoptosis, and massive infiltrates of T lymphocytes and macrophages. Low voltage electrochemotherapy with Taxol was shown to be more effective than surgical removal of the tumor with Taxol. CONCLUSIONS: These findings indicate that LEF-enhanced chemotherapy is an effective treatment of animals bearing metastatic melanoma. PMID- 12912971 TI - Epidermal growth factor receptor blockade potentiates apoptosis mediated by Paclitaxel and leads to prolonged survival in a murine model of oral cancer. AB - PURPOSE: Because survival for patients with oral cancer has not improved over the past 25 years, new approaches for treatment are needed. Targeted molecular therapy against epidermal growth factor receptor (EGFR) has shown promise as an adjuvant therapy in preliminary studies in several solid tumors, including head and neck cancer. The objective of this study was to determine the efficacy of paclitaxel and PKI166, a novel inhibitor of EGFR, against oral cavity cancer. EXPERIMENTAL DESIGN AND RESULTS: JMAR human oral cancer cells were pretreated for 1 h with PKI166 and then stimulated with epidermal growth factor. EGFR-specific tyrosine kinase autophosphorylation measured by Western immunoblotting was inhibited by PKI166 in a dose-dependent fashion at all doses tested (0.01-1 micro M). Next, the induction of apoptosis in JMAR cells treated with paclitaxel (0.001 to 0.1 micro M) with or without PKI166 (0, 1, or 2 micro M) was determined using a propidium iodide assay. The addition of 2.0 micro M PKI166 significantly increased tumor cell death, shifting the amount of paclitaxel needed to induce apoptosis in 50% of cells from 0.1 to 0.001 micro M. These in vitro findings were confirmed using an orthotopic model of oral cancer. JMAR oral cancer cells were implanted into the tongues of nude mice. After lingual tumors developed, mice were randomized into four groups (n = 10): (a) oral PKI166 (100 mg/kg); (b) i.p. paclitaxel (200 micro g/wk); (c) PKI166 and paclitaxel; or (d) placebo. Mice treated with PKI166/paclitaxel demonstrated a significant increase in survival (P = 0.028). After necropsy, all tongue tumors were evaluated for apoptosis by the terminal deoxynucleotidyl transferase-mediated nick end labeling assay. A greater apoptotic fraction of tumor cells was found in tumors of mice treated with paclitaxel and PKI166 as compared with the other treatment groups (136.4 versus 37.8; P = 0.016). CONCLUSIONS: Combination therapy with paclitaxel and PKI166 prolongs survival in an orthotopic preclinical model of tongue cancer by increasing programmed cell death of oral cancer. PMID- 12912973 TI - HER2 regulation of peroxisome proliferator-activated receptor gamma (PPARgamma) expression and sensitivity of breast cancer cells to PPARgamma ligand therapy. AB - Induction of terminal differentiation of cancer cells is an evolving novel therapeutic approach, and accordingly, peroxisome proliferator-activated receptor gamma (PPARgamma), a ligand-stimulated transcription factor with differentiation promoting activity and overexpressed in a variety of cancers, has emerged as one of the promising therapeutic targets. Because c-erbB family growth factor receptor 2 (HER2) overexpression is one of the most recognizable molecular dysfunctions in breast tumors, in the studies presented here, we explored the effect of HER2 overexpression on the status of PPARgamma expression and on the sensitivity of breast cancer cells to PPARgamma-ligand troglitazone-induced growth inhibition. We show that HER2 overexpression in MCF7 breast cancer cells enhanced the expression of PPARgamma-mRNA and -protein. Furthermore, PPARgamma expression was dramatically increased in 11 of 16 breast tumors as compared with the adjacent normal tissue. In addition, HER2 up-regulation resulted in a partial inhibition of transcriptional activity of the endogenous PPARgamma, stimulation to differentiation, and resistance to troglitazone-mediated inhibition of anchorage-independent growth of breast cancer cells. Conversely, down-regulation of HER2 by anti-HER2 monoclonal antibody Herceptin led to a decreased level of PPARgamma protein and sensitization of breast cancer cells to the inhibitory effects of troglitazone. In summary, these findings show for the first time that HER2 up-regulates PPARgamma expression and modulates the sensitivity of breast cancer cells to PPARgamma ligand therapy. PMID- 12912974 TI - Activation of a p53-mediated apoptotic pathway in quiescent lymphocytes after the inhibition of DNA repair by fludarabine. AB - PURPOSE: The inhibition of UV-initiated DNA repair by 9-beta-D-arabinofuranosyl-2 fluoroadenine (F-ara-A), the nucleoside of fludarabine, induces apoptosis in quiescent human lymphocytes. The sensing and signaling mechanisms after DNA repair inhibition by F-ara-A are unknown. The purpose of this study was 2-fold: (a) determine the importance of the inhibition of DNA repair processes for F-ara A cytotoxicity and (b) identify the apoptotic signaling mechanism(s) that respond to DNA repair inhibition by F-ara-A. EXPERIMENTAL DESIGN: Lymphocytes were treated with F-ara-A to accumulate the active triphosphate metabolite and subsequently DNA repair was activated by UV irradiation. Cell viability was quantitated with respect to the treatments alone and in combination to evaluate the actions of F-ara-A inhibition of DNA repair on p53 status and Fas death receptor ligand expression and function. RESULTS: Preincubation of lymphocytes with 3 micro M F-ara-A inhibited DNA repair initiated by 2 J/m(2) UV and induced greater than additive apoptosis after 24 h. After equivalent repair inhibition with 0.1 micro M aphidicolin, there was apparently lesser p53 activation and significantly less apoptosis in irradiated lymphocytes than after 3 micro M F-ara A. Blocking the incorporation of F-ara-A nucleotide into repairing DNA using 30 micro M aphidicolin lowered the apoptotic response to that observed with aphidicolin and UV. p53 serine 15 phosphorylation and protein accumulation were detected 2 h after treatment. Fas and Fas ligand mRNA expression and protein levels increased significantly after repair inhibition. Neutralizing antibodies against Fas or Fas ligand significantly reduced apoptosis. CONCLUSIONS: These results suggest that inhibition of UV-induced DNA repair by F-ara-A is critical for cytotoxicity and that induction of apoptosis may be conducted by a p53 mediated signaling mechanism to the Fas death pathway. PMID- 12912975 TI - Immunohistochemical variation of human equilibrative nucleoside transporter 1 protein in primary breast cancers. PMID- 12912976 TI - Zoledronic acid exhibits inhibitory effects on osteoblastic and osteolytic metastases of prostate cancer. PMID- 12912979 TI - Lipid-dependent recruitment of neuronal Src to lipid rafts in the brain. AB - Although most Src family tyrosine kinases are modified by palmitoylation as well as myristoylation, Src itself is only myristoylated. Dual acylation is important for attachment to liquid-ordered microdomains or lipid rafts. Accordingly, Src is excluded from lipid rafts in fibroblasts. Evidence of partial genetic redundancy between Src and Fyn for brain-specific targets suggests that these two kinases may occupy overlapping subcellular locations. Neuronal Src (NSrc), an alternative isoform of Src with a 6-amino acid insert in the Src homology 3 domain, is highly expressed in neurons. We investigated whether this structural difference in NSrc allows it to associate with lipid rafts. We found that perinatal mouse brains express predominantly NSrc, which is partly (10-20%) in a lipid raft fraction from brain but not fibroblasts. The association of Src with brain lipid rafts does not depend on the NSrc insert but depends on the amino-terminal myristoylation signal. A crude lipid fraction from brain promotes NSrc entry into rafts in vitro. Moreover, lipid raft-localized NSrc is more catalytically active than NSrc from the soluble fraction, possibly because raft localization alters access to other tyrosine kinases and phosphatases. These findings suggest that NSrc may be involved in signaling from lipid rafts in mouse brain. PMID- 12912978 TI - Cloning, expression, characterization, and role in autocrine cell growth of cell surface retention sequence binding protein-1. AB - Cell surface retention sequence binding protein-1 (CRSBP-1) is a cell surface binding protein for the cell surface retention sequence (CRS) motif of the v-sis gene product (platelet-derived growth factor-BB). It has been shown to be responsible for cell surface retention of the v-sis gene product in v-sis transformed cells (fibroblasts) and has been hypothesized to play a role in autocrine growth and transformation of these cells. Here we demonstrate that the CRSBP-1 cDNA cloned from bovine liver libraries encodes a 322-residue type I membrane protein containing a 23-residue signal peptide, a 215-residue cell surface domain, a 21-residue transmembrane domain, and a 63-residue cytoplasmic domain. CRSBP-1 expressed in transfected cells is an approximately 120-kDa disulfide-linked homodimeric glycoprotein and exhibits dual ligand (CRS containing growth regulators (v-sis gene product and insulin-like growth factor binding protein-3, IGFBP-3) and hyaluronic acid) binding activity. CRSBP-1 overexpression (by stable transfection of cells with CRSBP-1 cDNA) enhances autocrine loop signaling, cell growth, and tumorigenicity (in mice) of v-sis transformed cells. CRSBP-1 expression also enhances autocrine cell growth mediated by IGFBP-3 in human lung carcinoma cells (H1299 cells), which express very little, if any, endogenous CRSBP-1 and exhibits a mitogenic response to exogenous IGFBP-3, stably transfected with IGFBP-3 cDNA. However, CRSBP-1 overexpression does not affect growth of normal and transformed cells that do not produce these CRS-containing growth regulators. These results suggest that CRSBP 1 plays a role in autocrine regulation of cell growth mediated by growth regulators containing CRS. PMID- 12912980 TI - Differential contribution of inhibitory phosphorylation of CDC2 and CDK2 for unperturbed cell cycle control and DNA integrity checkpoints. AB - Inhibition of cyclin-dependent kinases (CDKs) by Thr14/Tyr15 phosphorylation is critical for normal cell cycle progression and is a converging event for several cell cycle checkpoints. In this study, we compared the relative contribution of inhibitory phosphorylation for cyclin A/B1-CDC2 and cyclin A/E-CDK2 complexes. We found that inhibitory phosphorylation plays a major role in the regulation of CDC2 but only a minor role for CDK2 during the unperturbed cell cycle of HeLa cells. The relative importance of inhibitory phosphorylation of CDC2 and CDK2 may reflect their distinct cellular functions. Despite this, expression of nonphosphorylation mutants of both CDC2 and CDK2 triggered unscheduled histone H3 phosphorylation early in the cell cycle and was cytotoxic. DNA damage by a radiomimetic drug or replication block by hydroxyurea stimulated a buildup of cyclin B1 but was accompanied by an increase of inhibitory phosphorylation of CDC2. After DNA damage and replication block, all cyclin-CDK pairs that control S phase and mitosis were to different degrees inhibited by phosphorylation. Ectopic expression of nonphosphorylated CDC2 stimulated DNA replication, histone H3 phosphorylation, and cell division even after DNA damage. Similarly, a nonphosphorylation mutant of CDK2, but not CDK4, disrupted the G2 DNA damage checkpoint. Finally, CDC25A, CDC25B, a dominant-negative CHK1, but not CDC25C or a dominant-negative WEE1, stimulated histone H3 phosphorylation after DNA damage. These data suggest differential contributions for the various regulators of Thr14/Tyr15 phosphorylation in normal cell cycle and during the DNA damage checkpoint. PMID- 12912981 TI - WITHDRAWN BY THE AUTHORS: Molecular mechanism of extracellular nucleotide-induced degranulation from human polymorphonuclear leukocytes: Role of leukotriene B4 and p38 MAP kinase as essential intermediates. AB - WITHDRAWN BY THE AUTHORS. PMID- 12912982 TI - Oligomerization of Ebola virus VP30 is essential for viral transcription and can be inhibited by a synthetic peptide. AB - Transcription of Ebola virus (EBOV)-specific mRNA is driven by the nucleocapsid proteins NP, VP35, and L. This process is further dependent on VP30, an essential EBOV-specific transcription factor. The present study addresses the self-assembly of VP30 and the functional significance of this process for viral transcription and propagation. Essential for oligomerization of VP30 is a region spanning amino acids 94-112. Within this region a cluster of four leucine residues is of critical importance. Mutation of only one of these leucine residues resulted in oligomerization-deficient VP30 molecules that were no longer able to support EBOV specific transcription. The essential role of homo-oligomerization for the function of VP30 was further corroborated by the finding that mixed VP30 oligomers consisting of VP30 and transcriptionally inactive VP30 mutants were impaired in their ability to support EBOV transcription. The dominant negative effect of these VP30 mutants was dependent on their ability to bind to VP30. The oligomerization of VP30 could be dose dependently inhibited by a 25-mer peptide (E30pep-wt) derived from the presumed oligomerization domain (IC50,1 mum). A control peptide (E30pep-3LA), in which three leucines were changed to alanine, had no inhibitory effect. Thus, E30pep-wt seemed to bind efficiently to VP30 and consequently blocked the oligomerization of the protein. When E30pep-wt was transfected into EBOV-infected cells, the peptide inhibited viral replication suggesting that inhibition of VP30 oligomerization represents a target for EBOV antiviral drugs. PMID- 12912983 TI - Two mammalian longevity assurance gene (LAG1) family members, trh1 and trh4, regulate dihydroceramide synthesis using different fatty acyl-CoA donors. AB - Overexpression of upstream of growth and differentiation factor 1 (uog1), a mammalian homolog of the yeast longevity assurance gene (LAG1), selectively induces the synthesis of stearoyl-containing sphingolipids in mammalian cells (Venkataraman, K., Riebeling, C., Bodennec, J., Riezman, H., Allegood, J. C., Sullards, M. C., Merrill, A. H. Jr., and Futerman, A. H. (2002) J. Biol. Chem. 277, 35642-35649). Gene data base analysis subsequently revealed a new subfamily of proteins containing the Lag1p motif, previously characterized as translocating chain-associating membrane (TRAM) protein homologs (TRH). We now report that two additional members of this family regulate the synthesis of (dihydro)ceramides with specific fatty acid(s) when overexpressed in human embryonic kidney 293T cells. TRH1 or TRH4-overexpression elevated [3H](dihydro)ceramide synthesis from l-[3-3H]serine and the increase was not blocked by the (dihydro)ceramide synthase inhibitor, fumonisin B1 (FB1). Analysis of sphingolipids by liquid chromatography electrospray tandem mass spectrometry revealed that TRH4 overexpression elevated mainly palmitic acid-containing sphingolipids whereas TRH1 overexpression increased mainly stearic acid and arachidic acid, which in both cases were further elevated upon incubation with FB1. A similar fatty acid specificity was obtained upon analysis of (dihydro)ceramide synthase activity in vitro using various fatty acyl-CoA substrates, although in a FB1-sensitive manner. Moreover, in homogenates from TRH4-overexpressing cells, sphinganine, rather than sphingosine was the preferred substrate, whereas no preference was seen in homogenates from TRH1-overexpressing cells. These findings lend support to our hypothesis (Venkataraman, K., and Futerman, A. H. (2002) FEBS Lett. 528, 3-4) that Lag1p family members regulate (dihydro)ceramide synthases responsible for production of sphingolipids containing different fatty acids. PMID- 12912984 TI - Molecular similarities in the ligand binding pockets of an odorant receptor and the metabotropic glutamate receptors. AB - The 5.24 odorant receptor is an amino acid sensing receptor that is expressed in the olfactory epithelium of fish. The 5.24 receptor is a G-protein-coupled receptor that shares amino acid sequence identity to mammalian pheromone receptors, the calcium-sensing receptor, the T1R taste receptors, and the metabotropic glutamate receptors (mGluRs). It is most potently activated by the basic amino acids L-lysine and L-arginine. In this study we generated a homology model of the ligand binding domain of the 5.24 receptor based on the crystal structure of mGluR1 and examined the proposed lysine binding pocket using site directed mutagenesis. Mutants of truncated glycosylated versions of the receptor containing only the extracellular domain were analyzed in a radioligand binding assay, whereas the analogous full-length membrane-bound mutants were studied using a fluorescence-based functional assay. In silico analysis predicted that aspartate 388 interacts with the terminal amino group on the side chain of the docked lysine molecule. This prediction was supported by experimental observations demonstrating that mutation of this residue caused a 26-fold reduction in the affinity for L-lysine but virtually no change in the affinity for the polar amino acid L-glutamine. In addition, mutations in four highly conserved residues (threonine 175, tyrosine 223, and aspartates 195 and 309) predicted to establish interactions with the alpha amino group of the bound lysine ligand greatly reduced or eliminated binding and receptor activation. These results define the essential features of amino acid selectivity within the 5.24 receptor binding pocket and highlight an evolutionarily conserved motif required for ligand recognition in amino acid activated receptors in the G protein-coupled receptor superfamily. PMID- 12912985 TI - Purification and characterization of Chinese hamster phosphatidylserine synthase 2. AB - Phosphatidylserine (PtdSer) in mammalian cells is synthesized through the action of PtdSer synthase (PSS) 1 and 2, which catalyze the conversion of phosphatidylcholine and phosphatidylethanolamine, respectively, to PtdSer. The PtdSer synthesis in intact cells and an isolated membrane fraction is inhibited by exogenous PtdSer, indicating that inhibition of PtdSer synthases by PtdSer is important for the regulation of PtdSer biosynthesis. In this study, to examine whether the inhibition occurs through the direct interaction of PtdSer with the synthases or is mediated by unidentified factor(s), we purified a FLAG and HA peptide-tagged form of Chinese hamster PSS 2 to near homogeneity. The purified enzyme, as well as the crude enzyme in a membrane fraction, was inhibited on the addition of PtdSer to the enzyme assay mixture. In contrast to PtdSer, phosphatidylcholine and phosphatidylethanolamine did not significantly inhibit the purified enzyme. Furthermore, PtdSer-resistant PtdSer synthesis was observed on cell-free assaying of the membrane fraction prepared from a Chinese hamster ovary cell strain whose PtdSer synthesis in vivo is not inhibited by exogenous PtdSer. These results suggested that the interaction of PtdSer with PSS 2 or a very minor protein co-purified with PSS 2 was critical for the regulation of PSS 2 activity in intact cells. PMID- 12912986 TI - Unexpected protein families including cell defense components feature in the N myristoylome of a higher eukaryote. AB - N-Myristoylation is an irreversible modification that affects the membrane binding properties of crucial cytoplasmic proteins from signal transduction cascades. We characterized the two putative N-myristoyltransferases of Arabidopsis thaliana as a means of investigating the entire N-myristoylation proteome (N-myristoylome) in a higher eukaryote. AtNMT1 compensated for the nmt1 defect in yeast, whereas AtNMT2 and chimeras of the two genes did not. Only AtNMT1 modified known N-myristoylated proteins in vitro. AtNMT1 is therefore responsible for the A. thaliana N-myristoylome, whereas AtNMT2 does not seem to have usual myristoylation activity. We began with the whole set of N myristoylated G proteins in the A. thaliana proteome. We then used a reiterative approach, based on the in vitro N-myristoylation of more than 60 different polypeptides, to determine the substrate specificity of AtNMT1. We found that the positive charge on residue 7 of the substrate was particularly important in substrate recognition. The A. thaliana N-myristoylome consists of 437 proteins, accounting for 1.7% of the complete proteome. We demonstrated the N myristoylation of several unexpected protein families, including innate immunity proteins, thioredoxins, components of the protein degradation pathway, transcription factors, and a crucial regulatory enzyme of glycolysis. The role of N-myristoylation is discussed in each case; in particular, this process may underlie the "guard" hypothesis of innate immunity. PMID- 12912987 TI - Lipid hydroperoxides activate the mitogen-activated protein kinase Mpk1p in Saccharomyces cerevisiae. AB - Saccharomyces cerevisiae is capable of responding to oxidants, including lipid peroxidation products. We investigate here the role of the mitogen-activated protein kinase Mpk1p in protection against linoleic acid hydroperoxide (LoaOOH), a product of radical attack on an unsaturated lipid. MPK1 was found to be required for resistance to LoaOOH. Furthermore, Mpk1p was rapidly and transiently phosphorylated in response to LoaOOH. This phosphorylation was dose-dependent and stimulated by sublethal concentrations as low as 1 mum in the external medium. Such low doses have been shown to result in resistance to subsequent challenge with a higher dose through the process of adaptation. However MPK1 was not essential for this adaptive response. MPK1 was also not involved in cell cycle modulation and acted independently of the cell cycle-regulating Oca1p. Transcriptional profiling of the mpk1Delta cells during LoaOOH stress indicated that Mpk1p may be important in effecting changes to the cell surface and metabolism during LoaOOH exposure. Furthermore, it revealed that Mpk1p is required for the regulation of 97 LoaOOH-responsive transcripts. Evidence is presented that the activation of Mpk1p may be caused by the activation of protein kinase C by LoaOOH. PMID- 12912988 TI - Brown adipose tissue Ca2+-ATPase: uncoupled ATP hydrolysis and thermogenic activity. AB - In this report a sarco/endoplasmic reticulum Ca2+-ATPase (SERCA) was identified in rats brown adipose tissue. Electrophoretic analysis of brown fat microssomal protein yields a 110-kDa band that is reactive to SERCA 1 antibody but is not reactive to SERCA 2 antibodies. Nevertheless, the kinetics properties of the brown fat SERCA differ from the skeletal muscle SERCA 1 inasmuch they manifest a different Ca2+ affinity and a much higher degree of ATPase/Ca2+ uncoupling. A SERCA enzyme is not found in white fat. Fatty acids promoted Ca2+ leakage from brown fat vesicles. The heat released during ATP hydrolysis was -24.7 kcal/mol when a Ca2+ gradient was formed across the vesicles membrane and -14.4 kcal/mol in the absence of a gradient. The data reported suggest that in addition to storing Ca2+ inside the endoplasmic reticulum, the Ca2+-ATPase may represent a source of heat production contributing to the thermogenic function of brown adipose tissue. PMID- 12912989 TI - Target of rapamycin (TOR)-signaling and RAIP motifs play distinct roles in the mammalian TOR-dependent phosphorylation of initiation factor 4E-binding protein 1. AB - The translational repressor protein eIF4E-binding protein 1 (4E-BP1, also termed PHAS-I) is regulated by phosphorylation through the rapamycin-sensitive mTOR (mammalian target of rapamycin) pathway. Recent studies have identified two regulatory motifs in 4E-BP1, an mTOR-signaling (TOS) motif in the C terminus of 4E-BP1 and an RAIP motif (named after its sequence) in the N terminus. Other recent work has shown that the protein raptor binds to mTOR and 4E-BP1. We show that raptor binds to full-length 4E-BP1 or a C-terminal fragment containing the TOS motif but not to an N-terminal fragment containing the RAIP motif. Mutation of several residues within the TOS motif abrogates binding to raptor, indicating that the TOS motif is required for this interaction. 4E-BP1 undergoes phosphorylation at multiple sites in intact cells. The effects of removal or mutation of the RAIP and TOS motifs differ. The RAIP motif is absolutely required for phosphorylation of sites in the N and C termini of 4E-BP1, whereas the TOS motif primarily affects phosphorylation of Ser-64/65, Thr-69/70, and also the rapamycin-insensitive site Ser-101. Phosphorylation of N-terminal sites that are dependent upon the RAIP motif is sensitive to rapamycin. The RAIP motif thus promotes the mTOR-dependent phosphorylation of multiple sites in 4E-BP1 independently of the 4E-BP1/raptor interaction. PMID- 12912990 TI - Cytosolic Arl2 is complexed with cofactor D and protein phosphatase 2A. AB - Arl2 is a member of the ADP-ribosylation factor family of 20-kDa GTPases that is highly conserved in eukaryotes. Recent results revealed that a portion of cellular Arl2 and its binding partner, BART, localize to mitochondria. Because approximately 90% of cellular Arl2 is cytosolic, we investigated properties of the soluble protein and found that it is stably bound in a complex that migrates in gel filtration medium with a predicted molecular mass of approximately 300 kDa. This complex was purified approximately 500-fold from the soluble fraction of bovine brain. Protein components were identified by mass spectroscopy and revealed the presence of four other proteins that include the tubulin folding cochaperone cofactor D and all three subunits of at least two protein phosphatase 2A (PP2A) protein phosphatase trimers. The presence of more than one PP2A B-type subunit and the low stoichiometry of Arl2 indicate that the purified preparation still contains a mixture of complexes that cannot currently be completely resolved. Thus, although all the soluble Arl2 in bovine brain is in high molecular mass complexes, only a portion of the total cellular cofactor D and PP2A are associated with the Arl2. We further show that the Arl2 in the complex cannot bind GTP and that complexed cofactor D does not efficiently participate in tubulin refolding reactions in a manner comparable with free cofactor D. Our data suggest functional roles for the cytosolic Arl2 complex in modulating tubulin and microtubule behavior as well as a possible role in apoptosis. PMID- 12912991 TI - Multiple roles for Arf6: sorting, structuring, and signaling at the plasma membrane. PMID- 12912992 TI - Rad51 recombinase and recombination mediators. PMID- 12912993 TI - Intron disruption of the annexin IV gene reveals novel transcripts. AB - Annexin IV (AIV), a Ca2+-dependent membrane-binding protein, is expressed in many epithelia. Annexin IV modifies membrane bilayers by increasing rigidity, reducing water and H+ permeability, promoting vesicle aggregation, and regulating ion conductances, all in a Ca2+-dependent manner. We have characterized a mouse in which a gene trap has been inserted into the first intron of annexin IV. Processing of the primary transcript is disrupted. Northern blot and immunoblot data indicated that annexin IV expression was eliminated in many but not all tissues. Immunohistochemical analysis, however, demonstrated that annexin IV expression was eliminated in some cell types, but was unaltered in others. 5' Rapid amplification of cDNA ends analysis of intestinal and kidney RNA revealed three transcripts, AIVa, AIVb, and AIVc. AIVa is widely distributed. AIVb is expressed only in the digestive tract. AIVc expression is very restricted. A selected number of epithelial cells of unique morphology demonstrate high concentrations. All three transcripts produce an identical annexin IV protein. The different tissue and cell-specific expression profiles of the three transcripts suggest that regulation of both the annexin IV gene expression and the cellular role of the protein are complex. The AIVa-/- mouse may become a valuable model to further study transcription and the physiological role of annexin IV. PMID- 12912994 TI - PB1 domains of MEKK2 and MEKK3 interact with the MEK5 PB1 domain for activation of the ERK5 pathway. AB - MEKK2 and MEKK3 are MAPK kinase kinases that activate the ERK5 pathway by phosphorylating and activating the MAPK kinase, MEK5. Activated MEK5 then phosphorylates and activates ERK5. PB1 domains were first defined in the p67phox and Bem1p proteins and have been shown to mediate protein-protein heterodimerization. A PB1 domain is encoded within the N-terminal portion of MEKK2, MEKK3, and MEK5. Herein, we analyze the functional role of MEKK2, MEKK3, and MEK5 PB1 domains in the ERK5 activation pathway. The PB1 domains of MEKK2 and MEKK3 bind the PB1 domain of MEK5 but do not significantly homo- or heterodimerize with one another in vitro. Furthermore, co-immunoprecipitation of MEKK2 and MEK5 from cell lysates shows that they form a complex in vivo. Deletion or mutation of the MEKK2 PB1 domain abolishes MEKK2-MEK5 complexes, demonstrating that the PB1 domain interaction is required for MEKK2-MEK5 interactions. Expression in cells of the MEKK2 or MEKK3 PB1 domain inhibits ERK5 activation, whereas expression of a mutant MEKK2 unable to bind the MEK5 PB1 domain or expression of the p67phox PB1 domain has no effect on ERK5 activation. These findings demonstrate that the PB1 domain mediates the association of MEKK2 and MEKK3 with MEK5 and that the respective PB1 domains of these kinases are critical for regulation of the ERK5 pathway. The free PB1 domain of MEKK2 or MEKK3 functions effectively to inhibit the ERK5 pathway but not the p38 or JNK pathways, demonstrating the specific and unique requirement of the MEKK2 and MEKK3 PB1 domain in regulating ERK5 activation. PMID- 12912995 TI - The effects of beta3 subunit incorporation on the pharmacology and single channel properties of oocyte-expressed human alpha3beta4 neuronal nicotinic receptors. AB - We compared the main properties of human recombinant alpha3beta4beta3 neuronal nicotinic receptors with those of alpha3beta4 receptors, expressed in Xenopus oocytes. beta3 incorporation decreased the channel mean open time (from 5.61 to 1.14 ms, after approximate correction for missed gaps) and burst length. There was also an increase in single channel slope conductance from 28.8 picosiemens (alpha3beta4) to 46.7 picosiemens (alpha3beta4beta3; in low divalent external solution). On the other hand, the calcium permeability (determined by a reversal potential method in chloride-depleted oocytes) and the pharmacological properties of beta3-containing receptors differed little from those of alpha3beta4. The main pharmacological difference in alpha3beta4beta3 "triplet" receptors was a 3-fold decrease in the potency of lobeline relative to acetylcholine. Nevertheless, there was no change in the rank order of potency for agonists (epibatidine >> lobeline > cytisine, 1,1-dimethyl-4-phenylpiperazinium iodide, nicotine > acetylcholine > carbachol for both receptors; measured at low agonist concentrations). Sensitivity to the competitive antagonists trimetaphan (0.2-1 microM) and dihydro-beta-erythroidine (30 microM) was similar for the two combinations, with a Schild KB for trimetaphan of 76 and 66 nM on alpha3beta4 and alpha3beta4beta3, respectively. The change in single channel conductance confirms that beta3 replaces a beta4 subunit in the pentamer. The absence of pronounced differences in the pharmacological profile of the triplet receptor argues against a role for the beta3 subunit in the formation of agonist binding sites, whereas the changes in channel kinetics suggest an important effect on receptor gating. The shortening of the burst length of beta3-containing receptors implies that any synaptic currents mediated by such channels would have faster decay kinetics. PMID- 12912996 TI - Sulfated polysaccharides enhance the biological activities of bone morphogenetic proteins. AB - Bone morphogenetic proteins (BMPs), which have been shown to be heparin-binding proteins, induce osteoblast differentiation in mesenchymal cells. In the present study, we examined the effects of heparin on the BMP activities in C2C12 myoblasts. Heparin dose dependently enhanced the osteoblast differentiation induced by not only homodimers of BMP-2 or BMP-4 but also heterodimers of BMP-2/6 or BMP-2/7. However, the osteoblast differentiation induced by the constitutively active BMPR-IA, a functional BMP type I receptor, was not affected by heparin. Heparan sulfate and dextran sulfate also enhanced the BMP-2 activity, although the chemically desulfated heparin-derivatives have lost this stimulatory capacity. Heparin dose-dependently suppressed the accumulation of BMP-2 from the culture media into the cell layer or BMPR-IA, and retained a large amount of BMP 2 in the culture media. The biological activity of BMP-2, which was evaluated using a BMP-responsive reporter gene expression, was prolonged in the presence of heparin. Taken together, these results suggest that sulfated polysaccharides enhance the biological activity of both homodimers and heterodimers of BMPs by continuously serving the ligands to their signaling receptors expressed on cell membranes. PMID- 12912997 TI - Human monocytes use Rac1, not Rac2, in the NADPH oxidase complex. AB - Phagocyte NADPH oxidase is critical for defense against pathogens and contributes to inflammatory tissue injury. One component of the NADPH oxidase complex is the small GTP-binding protein Rac. There are two isoforms of Rac, and Rac2 is the predominant isoform in neutrophils and has been shown to be essential for NADPH oxidase activity. In primary human monocytes we report that in contrast to neutrophils, Rac1 is the predominantly expressed isoform. Upon monocyte activation by a variety of agents, we found that Rac1 dissociates from Rho GDP dissociation inhibitor (RhoGDI) and translocates to the membrane. We also found that Rac1 interacts with two other NADPH oxidase components, p67phox and p47phox, upon monocyte activation. These data indicate that Rac1, and not Rac2, is a component of the activated NADPH oxidase in monocytes. This finding suggests that it may be possible to selectively interfere with either monocyte or neutrophil NADPH oxidase activity, thereby selectively targeting chronic versus acute inflammatory processes. PMID- 12912998 TI - Leukotriene D4 mediates survival and proliferation via separate but parallel pathways in the human intestinal epithelial cell line Int 407. AB - We demonstrated previously that leukotriene D4 (LTD4) regulates proliferation of intestinal epithelial cells through a CysLT receptor by protein kinase C (PKC)epsilon-dependent stimulation of the mitogen-activated protein kinase ERK1/2. Our current study provides the first evidence that LTD4 can activate 90 kDa ribosomal S6 kinase (p90RSK) and cAMP-responsive element-binding protein (CREB) via pertussis-toxin-sensitive Gi protein pathways. Transfection and inhibitor experiments revealed that activation of p90RSK, but not CREB, is a PKCepsilon/Raf-1/ERK1/2-dependent process. LTD4-mediated CREB activation was not affected by expression of kinase-dead p90RSK but was abolished by transfection with the regulatory domain of PKCalpha (a specific dominant-inhibitor of PKCalpha). Kinase-negative mutants of p90RSK and CREB (K-p90RSK and K-CREB) blocked the LTD4-induced increase in cell number and DNA synthesis (thymidine incorporation). Compatible with these results, flow cytometry showed that LTD4 caused transition from the G0/G1 to the S+G2/M cell cycle phase, indicating increased proliferation. Similar treatment of cells transfected with K-p90RSK resulted in cell cycle arrest in the G0/G1 phase, consistent with a role of p90RSK in LTD4-induced proliferation. On the other hand, expression of K-CREB caused a substantial buildup in the sub-G0/G1 phase, suggesting a role for CREB in mediating LTD4-mediated survival in intestinal epithelial cells. Our results show that LTD4 regulates proliferation and survival via distinct intracellular signaling pathways in intestinal epithelial cells. PMID- 12912999 TI - Dynamics of protamine 1 binding to single DNA molecules. AB - Protamine molecules bind to and condense DNA in the sperm of most vertebrates, packaging the sperm genome in an inactive state until it can be reactivated following fertilization. By using methods that enable the analysis of protamine binding to individual DNA molecules, we have monitored the kinetics of DNA condensation and decondensation by protamine 1 (P1) and synthetic peptides corresponding to specific segments of the bull P1 DNA binding domain. Our results show that the number of clustered arginine residues present in the DNA binding domain is the most important factor affecting the condensation and stability of the DNA-protamine complex prior to the formation of inter-protamine disulfide cross-links. The high affinity of P1 for DNA is achieved by the coordinated binding of three anchoring domains, which together in bull P1 contain 19 Arg residues. The single DNA molecule experiments show that sequences containing two or more anchoring domains have an off-rate that is at least 3 orders of magnitude slower than those containing a single domain. The use of Arg, rather than Lys residues, and the inclusion of Tyr or Phe residues in the hinge regions between anchoring domains provide additional stability to the complex. PMID- 12913000 TI - Association of transcription factor YY1 with the high molecular weight Notch complex suppresses the transactivation activity of Notch. AB - Notch receptors are evolutionarily conserved from Drosophila to human and play important roles in cell fate decisions. After ligand binding, Notch receptors are cleaved to release their intracellular domains. The intracellular domains, the activated form of Notch receptors, are then translocated into the nucleus where they interact with other transcriptional machinery to regulate the expression of cellular genes. To dissect the molecular mechanisms of Notch signaling, the cellular targets that interact with Notch1 receptor intracellular domain (N1IC) were screened. In this study, we found that endogenous transcription factor Ying Yang 1 (YY1) was associated with exogenous N1IC in human K562 erythroleukemic cells. The ankyrin (ANK) domain of N1IC and zinc finger domains of YY1 were essential for the association of N1IC and YY1 according to the pull-down assay of glutathione S-transferase fusion proteins. Furthermore, both YY1 and N1IC were present in a large complex of the nucleus to suppress the luciferase reporter activity transactivated by Notch signaling. The transcription factor YY1 indirectly regulated the transcriptional activity of the wild-type CBF1-response elements via the direct interaction of N1IC and CBF1. We also demonstrated the association between endogenous N1IC and intrinsic YY1 in human acute T-cell lymphoblastic leukemia cell lines. Taken together, these results indicate that transcription factor YY1 may modulate Notch signaling via association with the high molecular weight Notch complex. PMID- 12913001 TI - Cell entry of hepatitis C virus requires a set of co-receptors that include the CD81 tetraspanin and the SR-B1 scavenger receptor. AB - Several cell surface molecules have been proposed as receptor candidates, mediating cell entry of hepatitis C virus (HCV) on the basis of their physical association with virions or with soluble HCV E2 glycoproteins. However, due to the lack of infectious HCV particles, evidence that these receptor candidates support infection was missing. Using our recently described infectious HCV pseudotype particles (HCVpp) that display functional E1E2 glycoprotein complexes, here we show that HCV is a pH-dependent virus, implying that its receptor component(s) mediate virion internalization by endocytosis. Expression of the CD81 tetraspanin in non-permissive CD81-negative hepato-carcinoma cells was sufficient to restore susceptibility to HCVpp infection, confirming its critical role as a cell attachment factor. As a cell surface molecule likely to mediate endosomal trafficking, we demonstrate that the human scavenger receptor class B type 1 (SR-B1), a high-density lipoprotein-internalization molecule that we previously proposed as a novel HCV receptor candidate due to its affinity with E2 glycoproteins, is required for infection of CD81-expressing hepatic cells. By receptor competition assays, we found that SR-B1 antibodies that blocked binding of soluble E2 could prevent HCVpp infectivity. Furthermore, we establish that the hyper-variable region 1 of the HCV E2 glycoprotein is a critical determinant mediating entry in SR-B1-positive cells. Finally, by correlating expression of HCV receptors and infectivity, we suggest that, besides CD81 and SR-B1, additional hepatocyte-specific co-factor(s) are necessary for HCV entry. PMID- 12913002 TI - Crystal structure of trimeric carbohydrate recognition and neck domains of surfactant protein A. AB - Surfactant protein A (SP-A), one of four proteins associated with pulmonary surfactant, binds with high affinity to alveolar phospholipid membranes, positioning the protein at the first line of defense against inhaled pathogens. SP-A exhibits both calcium-dependent carbohydrate binding, a characteristic of the collectin family, and specific interactions with lipid membrane components. The crystal structure of the trimeric carbohydrate recognition domain and neck domain of SP-A was solved to 2.1-A resolution with multiwavelength anomalous dispersion phasing from samarium. Two metal binding sites were identified, one in the highly conserved lectin site and the other 8.5 A away. The interdomain carbohydrate recognition domain-neck angle is significantly less in SP-A than in the homologous collectins, surfactant protein D, and mannose-binding protein. This conformational difference may endow the SP-A trimer with a more extensive hydrophobic surface capable of binding lipophilic membrane components. The appearance of this surface suggests a putative binding region for membrane derived SP-A ligands such as phosphatidylcholine and lipid A, the endotoxic lipid component of bacterial lipopolysaccharide that mediates the potentially lethal effects of Gram-negative bacterial infection. PMID- 12913003 TI - Functional regulation of tissue plasminogen activator on the surface of vascular smooth muscle cells by the type-II transmembrane protein p63 (CKAP4). AB - We have demonstrated that tissue plasminogen activator (tPA) binds specifically to human vascular smooth muscle cells (VSMC) in a functionally relevant manner, both increasing plasminogen activation and decreasing tPA inhibition (Ellis, V., and Whawell, S. A. (1997) Blood 90, 2312-2322; Werner, F., Razzaq, T. M., and Ellis, V. (1999) J. Biol. Chem. 274, 21555-21561). To further understand this system we have now identified and characterized the protein responsible for this binding. Rat VSMC were surface-labeled with 125I, and cell lysates were subjected to an affinity chromatography scheme based on the previously identified tPA binding characteristics. A single radiolabeled protein of 63 kDa bound specifically and was eluted at low pH. This protein was isolated from large scale preparations of VSMC and unambiguously identified as the rat homologue of the human type-II transmembrane protein p63 (CKAP4) by matrix-assisted laser desorption ionization and nano-electrospray tandem mass spectrometry of tryptic fragments. In confirmation of this, a monoclonal antibody raised against authentic human p63 recognized the isolated protein in Western blotting. Immunofluorescence microscopy demonstrated that p63 was located principally in the endoplasmic reticulum but was also detected in significant quantities on the surface of human VSMC. In support of the hypothesis that p63 is the functional tPA binding site on VSMC, an anti-p63 monoclonal antibody was found to block tPA binding. Furthermore, heterologous expression of an N-terminally truncated mutant of p63, which targets exclusively to the plasma membrane, led to an increase in tPA-catalyzed plasminogen activation. Therefore, p63 on the surface of VSMC may contribute to the functional regulation of the plasminogen activation system in the vessel wall. PMID- 12913004 TI - The noncatalytic portion of human UDP-glucose: glycoprotein glucosyltransferase I confers UDP-glucose binding and transferase function to the catalytic domain. AB - The eukaryotic cell monitors the fidelity of protein folding in the endoplasmic reticulum and only permits properly folded and/or assembled proteins to transit to the Golgi compartment in a process termed "quality control." An endoplasmic reticulum (ER) lumenal sensor for quality control is the UDP-glucose:glycoprotein glucosyltransferase that targets unfolded glycoproteins for transient, calcium dependent glucosylation. This modification mediates glycoprotein interaction with the folding machinery comprised of calnexin or calreticulin in conjunction with ERp57. Two human UGT homologues, HUGT1 and HUGT2, exist that share 55% identity. The highest degree of identity resides in the COOH-terminal 20% of these proteins, the putative catalytic domain of HUGT1. However, only HUGT1 displays the expected functional activity. The contribution of the NH2-terminal remainder of HUGT1 to glucosyltransferase function is presently unknown. In this report we demonstrate that HUGT2 is localized to the ER in a manner that overlaps the distribution of HUGT1. Analysis of a series of HUGT1 and HUGT2 chimeric proteins demonstrated that the carboxyl-terminal region of HUGT2 contains a catalytic domain that is functional in place of the analogous portion of HUGT1. Whereas neither catalytic domain displayed detectable activity when expressed alone, co expression of either catalytic domain with the noncatalytic amino-terminal portion of HUGT1 conferred UDP-Glc binding and transfer of glucose that was specific for unfolded glycoprotein substrates. The results indicate that the amino-terminal 80% of HUGT1 is required for activation of the catalytic domain, whereas the homologous portion of HUGT2 cannot provide this function. PMID- 12913005 TI - Subunit arrangement in V-ATPase from Thermus thermophilus. AB - The V0V1-ATPase of Thermus thermophilus catalyzes ATP synthesis coupled with proton translocation. It consists of an ATPase-active V1 part (ABDF) and a proton channel V0 part (CLEGI), but the arrangement of each subunit is still largely unknown. Here we found that acid treatment of V0V1-ATPase induced its dissociation into two subcomplexes, one with subunit composition ABDFCL and the other with EGI. Exposure of the isolated V0 to acid or 8 m urea also produced two subcomplexes, EGI and CL. Thus, the C subunit (homologue of d subunit, yeast Vma6p) associates with the L subunit ring tightly, and I (homologue of 100-kDa subunit, yeast Vph1p), E, and G subunits constitute a stable complex. Based on these observations and our recent demonstration that D, F, and L subunits rotate relative to A3B3 (Imamura, H., Nakano, M., Noji, H., Muneyuki, E., Ohkuma, S., Yoshida, M., and Yokoyama, K. (2003) Proc. Natl. Acad. Sci. U. S. A. 100, 2312 2315; Yokoyama, K., Nakano, M., Imamura, H., Yoshida, M., and Tamakoshi, M. (2003) J. Biol. Chem. 278, 24255-24258), we propose that C, D, F, and L subunits constitute the central rotor shaft and A, B, E, G, and I subunits comprise the surrounding stator apparatus in the V0V1-ATPase. PMID- 12913007 TI - Blocking the function of tyrosine phosphatase SHP-2 by targeting its Src homology 2 domains. AB - SHP-2 is an Src homology 2 (SH2) domain-containing tyrosine phosphatase with crucial functions in cell signaling and major pathological implications. It stays inactive in the cytosol and is activated by binding through its SH2 domains to tyrosine-phosphorylated receptors on the cell surface. One such cell surface protein is PZR, which contains two tyrosine-based inhibition motifs responsible for binding of SHP-2. We have generated a glutathione S-transferase fusion protein carrying the tandem tyrosine-based inhibition motifs of PZR, and the protein was tyrosine-phosphorylated by co-expressing c-Src in Escherichia coli cells. The purified phosphoprotein displays a strong binding to SHP-2 and causes its activation in vitro. However, when introduced into NIH 3T3 cells by using a protein delivery reagent, it effectively inhibited the activation of ERK1/2 induced by growth factors and serum but not by phorbol ester, in reminiscence of the effects caused by expression of dominant negative SHP-2 mutants and deletion of functional SHP-2. The data suggest that the exogenously introduced PZR protein specifically binds SHP-2, blocks its translocation, and renders it functionally incompetent. This is further supported by the fact that the phosphorylated PZR protein had no inhibitory effects on fibroblasts derived from mice expressing only a mutant SHP-2 protein lacking most of the N-terminal SH2 domain. This study thus provides a novel and highly specific method to interrupt the function of SHP 2 in cells. PMID- 12913006 TI - Regulated intramembrane proteolysis of the p75 neurotrophin receptor modulates its association with the TrkA receptor. AB - The generation of biologically active proteins by regulated intramembrane proteolysis is a highly conserved mechanism in cell signaling. Presenilin dependent gamma-secretase activity is responsible for the intramembrane proteolysis of selected type I membrane proteins, including beta-amyloid precursor protein (APP) and Notch. A small fraction of intracellular domains derived from both APP and Notch translocates to and appears to function in the nucleus, suggesting a generic role for gamma-secretase cleavage in nuclear signaling. Here we show that the p75 neurotrophin receptor (p75NTR) undergoes presenilin-dependent intramembrane proteolysis to yield the soluble p75 intracellular domain. The p75NTR is a multifunctional type I membrane protein that promotes neurotrophin-induced neuronal survival and differentiation by forming a heteromeric co-receptor complex with the Trk receptors. Mass spectrometric analysis revealed that gamma-secretase-mediated cleavage of p75NTR occurs at a position located in the middle of the transmembrane (TM) domain, which is reminiscent of the amyloid beta-peptide 40 (Abeta40) cleavage of APP and is topologically distinct from the major TM cleavage site of Notch 1. Size exclusion chromatography and co-immunoprecipitation analyses revealed that TrkA forms a molecular complex together with either full-length p75 or membrane tethered C-terminal fragments. The p75-ICD was not recruited into the TrkA containing high molecular weight complex, indicating that gamma-secretase mediated removal of the p75 TM domain may perturb the interaction with TrkA. Independent of the possible nuclear function, our studies suggest that gamma secretase-mediated p75NTR proteolysis plays a role in the formation/disassembly of the p75-TrkA receptor complex by regulating the availability of the p75 TM domain that is required for this interaction. PMID- 12913008 TI - Unusual structural, functional, and stability properties of serine hydroxymethyltransferase from Mycobacterium tuberculosis. AB - From the genome analysis of the Mycobacterium tuberculosis two putative genes namely GlyA and GlyA2 have been proposed to encode for the enzyme serine hydroxymethyltransferase. We have cloned, overexpressed, and purified to homogeneity their respective protein products, serine hydroxymethyltransferase, SHM1 and SHM2. The recombinant SHM1 and SHM2 exist as homodimers of molecular mass about 90 kDa under physiological conditions, however, SHM2 has more compact conformation and higher thermal stability than SHM1. The most interesting structural observation was that the SHM1 contains 1 mol of pyridoxal 5'-phosphate (PLP)/mol of enzyme dimer. This is the first report of such a unique stoichiometry of PLP and enzyme dimer for SHMT. The SHM2 contains 2 mol of PLP/mol of enzyme dimer, which is the usual stoichiometry reported for SHMT. Functionally both the recombinant enzymes showed catalysis of reversible interconversion of serine and glycine and aldol cleavage of a 3-hydroxyamino acid. However, unlike SHMT from other sources both SHM1 and SHM2 do not undergo half-transamination reaction with d-alanine resulting in formation of apoenzyme but l-cysteine removed the prosthetic group, PLP, from both the recombinant enzymes leaving the respective inactive apoenzymes. Comparative structural studies on the two enzymes showed that the SHM1 is resistant to alkaline denaturation up to pH 10.5, whereas the native SHM2 dimer dissociates into monomer at pH 9. Urea- and guanidinium chloride-induced two-step unfolding of SHM1 and SHM2 with the first step being dissociation of dimer into apomonomer at low denaturant concentrations followed by unfolding of the stabilized monomer at higher denaturant concentrations. PMID- 12913009 TI - Effects of hydrogen peroxide upon nicotinamide nucleotide metabolism in Escherichia coli: changes in enzyme levels and nicotinamide nucleotide pools and studies of the oxidation of NAD(P)H by Fe(III). AB - DNA is damaged in vivo by the Fenton reaction mediated by Fe2+ and cellular reductants such as NADH, which reduce Fe3+ to Fe2+ and allow the recycling of iron. To study the response of Escherichia coli to such cycling, the activities of several enzymes involved in nicotinamide nucleotide metabolism were measured following an H2O2 challenge. NADPH-dependent peroxidase, NADH/NADP+ transhydrogenase, and glucose-6-phosphate dehydrogenase were most strongly induced, increasing 2.5-3-fold. In addition, the cellular ratios of NADPH to NADH increased 6- or 92-fold 15 min after exposure to 0.5 or 5 mm H2O2, respectively. In vitro, NADH was oxidized by Fe3+ up to 16-fold faster than NADPH, despite their identical reduction potentials. To understand this rate difference, the interactions of Fe3+ and Ga3+ with NAD(P)H were examined by 1H, 13C, and 31P NMR spectroscopy. Association with NADH occurred primarily with adenine at N7 and the amino group, but for NADPH, strong metal interactions also occurred at the 2' phosphate group. Interaction of M3+ (Fe3+ or Ga3+) with the adenine ring would bring it into close proximity to the redox-active nicotinamide ring in the folded form of NAD(P)H, but interaction of M3+ with the 2'-phosphate group would avoid this close contact. In addition, as determined by absorbance spectroscopy, the energy of the charge-transfer species was significantly higher for the Fe3+.NADPH complex than for the Fe3+.NADH complex. We therefore suggest that upon exposure to H2O2 the NADH pool is depleted, and NADPH, which is less reactive with Fe3+, functions as the major nicotinamide nucleotide reductant. PMID- 12913010 TI - Pemberton, sex and gender. PMID- 12913011 TI - Nutritional epidemiology--past, present, future. PMID- 12913012 TI - A compass for health: rethinking precaution and its role in science and public health. PMID- 12913013 TI - Malnutrition in England. 1934. PMID- 12913014 TI - 'Malnutrition in England' University College Hospital Magazine 1934. Some reflections in 2003 on the 1930s. PMID- 12913015 TI - Commentary: Minimum incomes for healthy living: then, now--and tomorrow? PMID- 12913016 TI - Commentary: The context and outcome of nutrition campaigning in 1934. PMID- 12913017 TI - Commentary: Socioeconomic inequalities and child growth. PMID- 12913018 TI - Commentary: John Pemberton professor of social medicine, Queen's University, Belfast, 1958-1976. PMID- 12913020 TI - Food insecurity, food choices, and body mass index in adults: nutrition transition in Trinidad and Tobago. AB - BACKGROUND: This study evaluated whether food insecurity and obesity were associated in a population sample in Trinidad. METHODS: A sample was drawn of 15 clusters of households, in north central Trinidad. Resident adults were enumerated. A questionnaire was administered including the short form Household Food Security Scale (HFSS). Heights and weights were measured. Analyses were adjusted for age, sex, and ethnic group. RESULTS: Data were analysed for 531/631 (84%) of eligible respondents including 241 men and 290 women with a mean age of 47 (range 24-89) years. Overall, 134 (25%) of subjects were classified as food insecure. Food insecurity was associated with lower household incomes and physical disability. Food insecure subjects were less likely to eat fruit (food insecure 40%, food secure 55%; adjusted odds ratio [OR] = 0.60, 95% CI: 0.36 0.99, P = 0.045) or green vegetables or salads (food insecure 28%, food secure 51%; adjusted OR = 0.46, 95% CI: 0.27-0.79, P = 0.005) on >/=5-6 days per week. Body mass index (BMI) was available for 467 (74%) subjects of whom 41 (9%) had BMI <20 kg/m(2), 157 (34%) had BMI 25-29 kg/m(2), and 120 (26%) had BMI >/=30 kg/m(2). Underweight (OR = 3.21, 95% CI: 1.17-8.81) was associated with food insecurity, but obesity was not (OR = 1.08, 95% CI: 0.55-2.12). CONCLUSIONS: Food insecurity was frequent at all levels of BMI and was associated with lower consumption of fruit and vegetables. Food insecurity was associated with underweight but not with present obesity. PMID- 12913021 TI - Commentary: Assessing food insecurity in Trinidad and Tobago. PMID- 12913022 TI - The World Health Organization Global Database on Child Growth and Malnutrition: methodology and applications. AB - BACKGROUND: For decades nutritional surveys have been conducted using various definitions, indicators and reference populations to classify child malnutrition. The World Health Organization (WHO) Global Database on Child Growth and Malnutrition was initiated in 1986 with the objective to collect, standardize, and disseminate child anthropometric data using a standard format. METHODS: The database includes population-based surveys that fulfil a set of criteria. Data are checked for validity and consistency and raw data sets are analysed following a standard procedure to obtain comparable results. Prevalences of wasting, stunting, under- and overweight in preschool children are presented using z scores based on the National Center for Health Statistics (NCHS)/WHO international reference population. New surveys are included on a continuous basis and updates are published bimonthly on the database's web site. RESULTS: To date, the database contains child anthropometric information derived from 846 surveys. With 412 national surveys from 138 countries and 434 sub-national surveys from 155 countries, the database covers 99% and 64% of the under 5 year olds in developing and developed countries, respectively. This wealth of information enables international comparison of nutritional data, helps identifying populations in need, evaluating nutritional and other public health interventions, monitoring trends in child growth, and raising political awareness of nutritional problems. CONCLUSIONS: The 15 years experience of the database can be regarded as a success story of international collaboration in standardizing child growth data. We recommend this model for monitoring other nutritional health conditions that as yet lack comparable data. PMID- 12913023 TI - Lack of association between tea and cardiovascular disease in college alumni. AB - BACKGROUND: Epidemiological studies suggest that tea intake, a major dietary source of flavonoids, may be associated with a decreased risk of cardiovascular disease (CVD). METHODS: We prospectively followed 17 228 subjects (mean age, 59.5 years) initially free of CVD and cancer from the College Alumni Health Study. Participants provided baseline self-reports of tea consumption (cups/day) and coronary risk factors. During a median follow-up of 15 years, there were 3372, 2615, and 757 cases of CVD, coronary heart disease (CHD), and stroke, respectively, ascertained from either self-reports or death certificates. RESULTS: Overall, the median level of tea consumption was 1 cup/day. Compared with participants consuming no tea, the multivariate relative risks (RR) of CVD for those drinking <1, 1, 2, 3, and >/=4 cups/day were 0.99, 0.96, 0.95, 0.91, and 0.95, respectively (P, trend = 0.19). The multivariate RR were 0.97, 0.98, 0.93, 0.85, and 0.98 for CHD (P, trend = 0.25), and 1.05, 0.89, 1.00, 1.09, and 0.83 for stroke (P, trend = 0.53). There was no evidence of effect modification. Changes in tea intake were assessed in a subgroup of 7730 men, with those continuing to drink tea having a non-significant 33% reduction in the risk of stroke. CONCLUSIONS: Tea intake, likely consumed as black tea, was not strongly associated with a reduced risk of CVD in this population of US college alumni. PMID- 12913024 TI - Commentary: This study failed? PMID- 12913025 TI - Intake of animal products and stroke mortality in the Hiroshima/Nagasaki Life Span Study. AB - BACKGROUND: To determine whether intake of animal products was associated with a reduced risk of stroke mortality in a large-scale population-based cohort in Japan. METHODS: A self-administered questionnaire, including questions on dietary habits, was mailed to the members of the Life Span Study, a cohort of people exposed and non-exposed to atomic bomb radiation, who were alive as of 1 September 1979. Animal products included frequency intake of beef/pork, chicken, ham/sausage, milk, dairy products, eggs, fish, and broiled fish. Responses were obtained from 40 349 people (72%): 15 350 men (mean age 54 years) and 24 999 women (mean age 58 years). The subjects were followed for 16 years, and deaths were ascertained by linkage to the nationwide family registration system of Japan. The associations between diet and stroke mortality were examined using a Cox proportional hazard model. RESULTS: During the follow-up period, 1462 stroke deaths occurred. Four animal products comprising eggs, dairy products, fish, and broiled fish were independently associated with a decreased risk of stroke mortality; while beef/pork, chicken, ham/ sausage, and milk consumption were not associated with stroke death. A composite measure of eggs, dairy products, fish, and broiled fish intake was calculated, and the highest tertile was significantly inversely associated with total stroke mortality (Hazards Ratio [HR] = 0.80, 95% CI: 0.68, 0.93) compared with the lowest tertile. The protective effect of animal product intake on total stroke death was largely confined to intracerebral haemorrhage death; the RH of intracerebral haemorrhage death for the highest tertile of consumption was 0.72 (95% CI: 0.53-0.98) compared with the lowest tertile; animal products intake was not related to cerebral infarction mortality (HR = 0.84; 95% CI: 0.67-1.06). CONCLUSIONS: Intake of animal products such as eggs, dairy products, and fish may be protective against intracerebral haemorrhage, but is not related to cerebral infarction mortality. PMID- 12913026 TI - Commentary: Protection from stroke by eating animal foods? Surely not! PMID- 12913027 TI - Demographics, lifestyles, health characteristics, and dietary intake among dietary supplement users in Japan. AB - BACKGROUND: The associations between supplement use and certain demographics, lifestyles, health characteristics, and dietary intakes have not been studied in a large population in non-Western societies. The objective of our study was to investigate the association between supplement use and demographics, lifestyles, health characteristics, and dietary intake in a population-based cohort study in Japan. METHODS: Subjects were the 78 531 participants (45-74 years) who completed a self-administered questionnaire in 1995 or 1998 in a 5-year follow-up survey by the Japan Public Health Center-based prospective Study on cancer and cardiovascular disease. The questionnaire included enquiries about supplement use, occupation, height, weight, smoking, alcohol, physical activity, dietary behaviours, working hours, subjective stress, as well as intakes for 138 foods. RESULTS: The supplement users were likely to have formerly smoked or never smoked. Female supplement users were likely to consume alcohol moderately. The prevalence of users was higher in the elderly, the self-employed, those with lower body mass index, greater physical activity, lower frequency of eating prepared food, higher frequency of eating out, and higher stress level in both sexes after mutual adjustment. Mean intakes of energy and nutrients were lower for users than for non-users. CONCLUSION: The demographics, lifestyles, health characteristics, and dietary intakes may need to be adjusted when evaluating the effect of dietary supplements on disease because they can become potential confounding factors. PMID- 12913028 TI - Commentary: Vitamin supplement use and confounding by lifestyle. PMID- 12913029 TI - Determinants of aflatoxin exposure in young children from Benin and Togo, West Africa: the critical role of weaning. AB - BACKGROUND: Dietary exposure to high levels of the fungal toxin, aflatoxin, occurs in West Africa, where long-term crop storage facilitates fungal growth. METHODS: We conducted a cross-sectional study in Benin and Togo to investigate aflatoxin exposure in children around the time of weaning and correlated these data with food consumption, socioeconomic status, agro-ecological zone of residence, and anthropometric measures. Blood samples from 479 children (age 9 months to 5 years) from 16 villages in four agro-ecological zones were assayed for aflatoxin-albumin adducts (AF-alb) as a measure of recent past (2-3 months) exposure. RESULTS: Aflatoxin-albumin adducts were detected in 475/479 (99%) children (geometric mean 32.8 pg/mg, 95% CI: 25.3-42.5). Adduct levels varied markedly across agro-ecological zones with mean levels being approximately four times higher in the central than in the northern region. The AF-alb level increased with age up to 3 years, and within the 1-3 year age group was significantly (P = 0.0001) related to weaning status; weaned children had approximately twofold higher mean AF-alb adduct levels (38 pg AF-lysine equivalents per mg of albumin [pg/mg]) than those receiving a mixture of breast milk and solid foods after adjustment for age, sex, agro-ecological zone, and socioeconomic status. A higher frequency of maize consumption, but not groundnut consumption, by the child in the preceding week was correlated with higher AF-alb adduct level. We previously reported that the prevalence of stunted growth (height for age Z-score HAZ) and being underweight (weight for age Z-score WAZ) were 33% and 29% respectively by World Health Organziation criteria. Children in these two categories had 30-40% higher mean AF-alb levels than the remainder of the children and strong dose- response relationships were observed between AF-alb levels and the extent of stunting and being underweight. CONCLUSIONS: Exposure to this common toxic contaminant of West African food increases markedly following weaning and exposure early in life is associated with reduced growth. These observations reinforce the need for aflatoxin exposure intervention strategies within high-risk countries, possibly targeted specifically at foods used in the post-weaning period. PMID- 12913030 TI - Cholesterol, coronary heart disease, and stroke in the Asia Pacific region. AB - BACKGROUND: Cholesterol levels in many Asian countries are rising. Predictions of the likely effects of this on the incidence of cardiovascular diseases have mostly relied on data from Western populations. Whether the associations between total cholesterol and cardiovascular diseases are similar in Asia is not established. METHODS: The Asia Pacific Cohort Studies Collaboration (APCSC) is an individual-participant data meta-analysis of prospective studies from the Asia Pacific region. Cox models were applied to the combined data from 29 cohorts to estimate the region-, sex-, and age-specific hazard ratios of major cardiovascular diseases by the fifths of total cholesterol. RESULTS: At baseline, the age/sex-adjusted mean value of total cholesterol was higher in Australia and New Zealand (ANZ) (5.52 +/- 1.05 mmol/l) than in Asia (4.87 +/- 1.05 mmol/l). During 2 million person-years of follow-up among 352 033 individuals, 4841 cardiovascular deaths were recorded. The association of total cholesterol with coronary heart disease and stroke was similar in Asian and ANZ cohorts. Overall, each 1-mmol/l higher level of total cholesterol was associated with 35% (95% CI: 26-44%) increased risk of coronary death, 25% (95% CI: 13-40%) increased risk of fatal or non-fatal ischaemic stroke, and 20% (95% CI: 8-30%) decreased risk of fatal haemorrhagic stroke. CONCLUSIONS: In both Asian and non-Asian populations in the Asia-Pacific region, total cholesterol is similarly strongly associated with the risk of CHD and ischaemic, but not haemorrhagic, stroke. Rising population-wide levels of cholesterol would be expected to contribute to a substantial increase in the overall burden of cardiovascular diseases in this region. PMID- 12913031 TI - Limited Pap screening associated with reduced risk of cervical cancer in South Africa. AB - BACKGROUND: We investigated the effect of Pap smear screening on the incidence of invasive cancer of the cervix in the Western Cape, South Africa where screening is limited. METHODS: Data were derived from a case-control study of the association of hormonal contraceptives and invasive cervical cancer. Incident cases (n = 524) of invasive cervical cancer who presented at two tertiary hospitals and controls (n = 1540) series matched for age, race, and place of residence were interviewed. Information on a wide range of variables was collected including whether the women had previously had a Pap smear taken and the number and timing of smears. Odds ratios (OR) and 95% CI were calculated using multiple logistic regression. RESULTS: The OR of cervical cancer was reduced among women who had ever had a smear (OR = 0.3, 95% CI: 0.3-0.4). The OR declined with increasing number of smears to 0.2 for >/=>3 smears (trend P = 0.0003). Among women who had a smear <5 years previously the OR was 0.3, but even if the smear was taken >/=15 years previously the women remained at reduced risk (OR = 0.5). CONCLUSION: The data suggest that even limited Pap smear screening reduces the risk of cervical cancer. Should a screening programme be successfully implemented, the incidence of cervical cancer might be reduced by as much as 70%. PMID- 12913032 TI - Commentary: Case-control studies of screening should carry a health warning. PMID- 12913034 TI - Iodine deficiency, radiation dose, and the risk of thyroid cancer among children and adolescents in the Bryansk region of Russia following the Chernobyl power station accident. AB - BACKGROUND: Little is known about the joint effect of iodine deficiency and radiation exposure on the risk of thyroid cancer. No epidemiological studies have been published assessing the modifying effect of iodine deficiency on radiation induced thyroid cancer following the Chernobyl accident. METHODS: A population sample of 3070 individuals (2590 ages 6-18, and 480 adults) from 75 settlements in the most highly contaminated regions of the Bryansk Oblast of Russia was identified and sampled for urinary iodine measurements in 1996, and iodine deficiency in each geopolitical unit (raion) was estimated. All cases of thyroid cancer were identified in those born 1968-1986 who were resident in the study area in May-June 1986 (34 histologically confirmed cases). The risk of thyroid cancer was examined in relation to population estimates of thyroid radiation dose and urinary iodine excretion level. RESULTS: The excess relative risk (ERR) of thyroid cancer was significantly associated with increasing thyroid radiation dose, and was inversely associated with urinary iodine excretion levels. There was a joint effect of radiation exposure and iodine deficiency. At 1 gray (Gy), the ERR in territories with severe iodine deficiency was approximately two times that in areas of normal iodine intake. CONCLUSIONS: These findings suggest that elimination of iodine deficiency in areas affected by Chernobyl may be important in reducing the effects of radiation exposure on the thyroid. If confirmed by studies based on individuals, they may have implications for the use of stable iodine in the case of population exposure to radioactive iodine. PMID- 12913033 TI - Aggregation of stomach cancer history in parents and offspring in comparison with other sites. AB - BACKGROUND: The aim of this study is to evaluate the magnitude of the aggregation of a stomach cancer history in parents and their offspring in comparison with that of a history at other sites. METHODS: We used the baseline data from the Japan Collaborative Cohort Study for Evaluation of Cancer Risk (JACC Study), which was initiated during 1988-1990 in Japan. Association of the cancer history of the subjects' parents with that of the subjects themselves and any of the subjects' siblings was evaluated with odds ratios (OR) by the crude and generalized estimating equations (GEE) technique for four sites: stomach, colorectum, liver, and lung/bronchus. RESULTS: The aggregation of a history of stomach cancer between parents and their offspring was evident with significant OR >2.5. The magnitude of the parent-offspring association of a disease history of the colorectum and liver was found to be greater than that for stomach cancer. Conversely, lung and bronchus cancer failed to demonstrate a significant aggregation. CONCLUSIONS: The hereditary and environmental influences shared by parents and offspring are likely to play a strong aetiological role in colorectal or liver cancer versus a weaker but still significant role in stomach cancer. In contrast, the aetiological role of familial predisposition to lung cancer was indeterminate, which suggests a predominant role of non-familial factors in the development of lung cancer. PMID- 12913035 TI - Pulmonary function as a predictor of lung cancer mortality in continuing cigarette smokers and in quitters. AB - BACKGROUND: Forced expiratory volume in 1 second (FEV(1)) may be useful for identifying smokers at higher risk of lung cancer. We examined the association of FEV(1) with lung cancer mortality (LCM) among cigarette smokers in the Multiple Risk Factor Intervention Trial (MRFIT). METHODS: In all, 6613 MRFIT baseline smokers alive at trial end in 1982 had acceptable FEV(1) measures and complete smoking history; men were classified as during-trial long-term quitters (N = 1292), intermittent quitters (1961), and never quitters (3360). Proportional hazards models for LCM were fit with quintiles of average FEV(1), adjusted for age, height, race, smoking history, and other risk factors. RESULTS: For long term, intermittent, and never quitters respectively, mean baseline cigarettes/ day was 28, 32, and 35; trial-averaged FEV(1) was 3201, 3146, and 3082 ml; and average decline in FEV(1) was -46.0, -54.6, and -62.5 ml/year. With median post trial mortality follow-up of 18 years, there were 363 lung cancer deaths. Age adjusted LCM rates varied across FEV(1) quintiles from 50 (lowest quintile) to 11 (highest quintile), 58 to 11, and 76 to 20, per 10 000 person-years, for long term quitters, intermittent quitters, and never quitters, respectively. Multivariate adjusted hazard ratios for 100 ml higher FEV(1) were 0.92 [P = 0.004], 0.95 [P = 0.003], and 0.95 [P < 0.0001] respectively. CONCLUSIONS: These results demonstrate the strong predictive value of FEV(1) for lung cancer among cigarette smokers independent of smoking history; results did not differ by during-trial quit status. FEV(1) may be a biological marker for smoking dose or it may be that genetic susceptibilities to both decreased FEV(1) and lung cancer are associated. PMID- 12913036 TI - Association of the metabolic syndrome with both vigorous and moderate physical activity. AB - BACKGROUND: Cross-sectional relationships between moderate and vigorous physical activity and the metabolic syndrome (MS) were examined in the Whitehall II study of civil servants (age 45-68 years). We assessed cardiovascular fitness and body mass index (BMI) as possible mediators of the observed association. METHODS: Measures of 2-hour glucose, systolic blood pressure, fasting triglycerides, waist hip ratio, and high density lipoprotein (HDL) cholesterol were obtained in 5153 white European participants. Participants in the most adverse sex-specific quintile for three or more of these risk factors were classified as having MS. Self-reported leisure-time physical activity was categorized into separate moderate and vigorous activity classes. BMI and resting heart rate (HR) were used to estimate body fatness and cardiovascular fitness respectively. RESULTS: The odds ratios (95% CI) for having the metabolic syndrome in the top categories of vigorous and moderate activity were 0.52 (95% CI: 0.40, 0.67) and 0.78 (95% CI: 0.63, 0.96) respectively, adjusted for age, sex, smoking, alcohol intake, socioeconomic status, and other activity. Adjustment for BMI and resting HR substantially attenuated both of the above associations. CONCLUSIONS: Moderate and vigorous physical leisure-time activity are each associated with reduced risk of being classified with MS independently of age, smoking, and high alcohol intake. Both vigorous and moderate activities may be beneficial to the MS cluster of risk factors among middle-aged populations. Reduced BMI and increased cardiovascular fitness may be important mediators of this association for both intensities of activity. PMID- 12913037 TI - Family study of the relationship between height and cardiovascular risk factors in the STANISLAS cohort. AB - BACKGROUND: Short stature is a risk factor for coronary heart disease and is associated with an adverse cardiovascular profile. Mechanisms responsible for this association remain unknown. A genetic contribution to this association would imply a familial clustering between height and cardiovascular risk factors. METHODS: This study investigated whether lipids and blood pressure (BP) levels shared a common familial component with height. The sample included 865 nuclear families from the French STANISLAS cohort volunteering for a free health examination between 1993 and 1994. Within-individual correlations and familial intra-trait and cross-trait correlations were estimated using the Estimating Equation technique extended to a bivariate phenotype. RESULTS: Height negatively correlated to total and low density lipoprotein cholesterol (LDL-C) and triglycerides in both parents and offspring, and positively correlated to high density lipoprotein cholesterol (HDL-C) in parents only. In offspring, the correlation between height and HDL-C markedly increased with sexual maturation to reach after puberty the same value as in parents. The correlation of height with systolic BP was negative in fathers and positive in sons, whereas it was non significant in mothers and daughters. The pattern of cross-trait familial correlations between height and LDL-C was compatible with the existence of a weak transmissible component explaining the relationship between these two traits. By contrast, the pattern observed for HDL-C and triglycerides was rather compatible with the influence of shared environmental factors. No familial clustering between height and BP levels was detected. CONCLUSIONS: The association between short stature and increased LDL-C might be partly of familial origin. PMID- 12913038 TI - Commentary: Disentangling the association between short height and cardiovascular risk--genes or environment? PMID- 12913039 TI - Improved methods for estimating incidence from linked hospital morbidity data. AB - BACKGROUND: Linked hospital morbidity data can be used to estimate the incidence of serious chronic disease. However, incidence rates calculated from first-time hospital admissions tend to be overestimated as a result of the erroneous inclusion of prevalent cases that have had previous hospital admissions prior to the study observation period. To address this problem, we have developed the backcasting method. METHOD: A retrograde survival model was implemented to calculate the level of over-ascertainment of incidence according to the number of years of linked data on which the estimates were based and corresponding correction factors were calculated. The method is illustrated using the example of linked hospital morbidity data on diabetes mellitus and then acute myocardial infarction, which was validated against the Perth MONICA database for cardiovascular disease. RESULTS: Corrected estimates of the incidence of diabetes and acute myocardial infarction were produced. The incidence of diabetes was shown to be lower than in North America in accordance with prevalence estimates, whereas the incidence of acute myocardial infarction was overestimated by approximately 10%. CONCLUSION: A new method is presented for estimating incidence trends in disease from linked hospital morbidity data. The advantages of this method are its ease of use with routinely collected data and the relatively low cost of applying it in comparison with community surveys or maintaining formal disease registers. The method has other applications using linked data, such as the study of trends in first-time health care procedures and pharmaceutical prescriptions. PMID- 12913040 TI - Birthweight and perinatal mortality: paradoxes, social class, and sibling dependencies. AB - BACKGROUND: Birthweight distributions among second-born infants depend on the birthweights of older siblings, with implications for weight-specific perinatal mortality. We wanted to study whether these relations were explained by socioeconomic levels, and to study time trends in a situation with decreasing perinatal mortality rates. METHODS: Births in the Norwegian Medical Birth Registry from 1967 to 1998 were linked to their mothers through their national identification numbers. The study population was 546 688 mothers with at least two singletons weighing >/==" BORDER="0">500 g at birth. Weight-specific perinatal mortality for second-born siblings in families with first-born siblings in either the highest or the lowest birthweight quartile was analysed. Maternal education and cohabitation status were used as measures of socioeconomic level. RESULTS: For all 500-g categories below 3500 g, mortality rates were significantly higher among second-born infants with an older sibling in the highest rather than the lowest weight quartile. This pattern was the same across three educational levels. The exclusion of preterm births did not change the effect pattern. A comparison of perinatal mortality among second siblings in terms of relative birthweight (z-scores) showed a reversal of the relative risks, although these were only significantly different from unity for the smallest infants. Conclusion The crossover in weight-specific perinatal mortality for second siblings by weight of first sibling is largely independent of socioeconomic level, and is not weakened by the decreasing perinatal mortality rates in the population over time. Family data should be taken into consideration when evaluating the risk of adverse pregnancy outcome relating to weight. PMID- 12913041 TI - Commentary: On the paradoxes of birthweight. PMID- 12913042 TI - A randomized trial of opinion leader endorsement in a survey of orthopaedic surgeons: effect on primary response rates. AB - BACKGROUND: Opinion leaders have been shown to have significant influence on the practice of health professionals and patient outcomes. METHODS: Using focus groups, key informants, and sampling to redundancy techniques, we developed a questionnaire of surgeons' preferences in the treatment of tibial shaft fractures. Twenty-two well-respected and widely known orthopaedic traumatologists endorsed the questionnaire. We randomized 395 surgeon members of the Orthopaedic Trauma Association to receive either a questionnaire that included a letter informing them of the opinion leaders' endorsement, or a questionnaire without the endorsement. RESULTS: Surgeons who received the letter of endorsement had a significantly lower response rate at 2, 4, and 8 weeks. The absolute difference in response rates was 7.8% (4.6% versus 12.4%, P < 0.05) at 2 weeks, 13.1% at 4 weeks (28.6% versus 41.7% P < 0.02), and 12.3% at 8 weeks (47.5% versus 59.8% P = 0.02). CONCLUSIONS: The addition of a letter listing expert surgeons who endorse the survey lead to significantly lower primary response rates. Those interested in influencing physician responses cannot always assume a positive effect from endorsement by opinion leaders PMID- 12913043 TI - Commentary: Improving response rates to mailed surveys: what do we learn from randomized controlled trials? PMID- 12913044 TI - Components of small area variation in fertility rates among married women in south India. AB - OBJECTIVE: To adapt and develop a method for finding out what proportion of the variation among small areas in the number of births to married women is excess (systematic) variation over and above the chance (random) variation. METHODS: We adopted a two-stage sampling procedure to select 20 sub-centres in south India. We contacted all households and collected information on recent births and socioeconomic variables from all ever married women aged 15-49 years. Systematic and random components of variance were estimated using Poisson regression, adjusting for socioeconomic factors. RESULTS: Of the observed variance in fertility, 65% is explained by the estimated systematic variation. Though many important explanatory variables are considered, the systematic variance is significant mainly among illiterate women, those aged under 18 years at marriage, the marginalized population, and those with labourer husbands. CONCLUSIONS: Poisson regression can be adapted to estimate the random and systematic components of variation in fertility among small areas. The systematic component of variance can further be adjusted for socioeconomic factors. Identification of the significant predictors will help health planners develop necessary interventions at the small area level. PMID- 12913045 TI - Relationship between body mass and gastro-oesophageal reflux symptoms: The Bristol Helicobacter Project. AB - OBJECTIVE: To examine the relationship between body mass and gastro-oesophageal reflux in a large population-representative sample from the UK. DESIGN AND SETTING: Cross-sectional population-based study, as part of a randomized controlled trial of eradication of Helicobacter pylori infection, in Southwest England. Subjects In all, 10 537 subjects, aged 20-59 years, were recruited from seven general practices. Subjects provided data on frequency and severity of dyspeptic symptoms and anthropometric measurements were taken. MAIN OUTCOME MEASURE: Relationship between overweight (body mass index [BMI] >/=25 kg/m(2) and 30 kg/m(2)) and frequency and severity of heartburn and acid regurgitation. RESULTS: Body mass index was strongly positively related to the frequency of symptoms of gastro-oesophageal reflux. The adjusted odds ratios (OR) for frequency of heartburn and acid regurgitation occurring at least once a week in overweight participants compared with those of normal weight were 1.82 (95% CI: 1.33-2.50) and 1.50 (95% CI: 1.13-1.99) respectively. Corresponding OR (95% CI) relating to obese patients were 2.91 (95% CI: 2.07-4.08) and 2.23 (95% CI: 1.44-3.45) respectively. The OR for moderate to severe reflux symptoms were raised in overweight and obese subjects but not to the same extent as frequency of symptoms and only the relationship between obesity and severity of heartburn reached conventional statistical significance: OR = 1.19; 95% CI: 1.07-1.33. CONCLUSIONS: Being above normal weight substantially increases the likelihood of suffering from heartburn and acid regurgitation and obese people are almost three times as likely to experience these symptoms as those of normal weight. PMID- 12913046 TI - Commentary: The relation of obesity, reflux and its implications. PMID- 12913047 TI - Genders, sexes, and health: what are the connections--and why does it matter? AB - Open up any biomedical or public health journal prior to the 1970s, and one term will be glaringly absent: gender. Open up any recent biomedical or public health journal, and two terms will be used either: (1) interchangeably, or (2) as distinct constructs: gender and sex. Why the change? Why the confusion?-and why does it matter? After briefly reviewing conceptual debates leading to distinctions between 'sex' and 'gender' as biological and social constructs, respectively, the paper draws on ecosocial theory to present 12 case examples in which gender relations and sex-linked biology are singly, neither, or both relevant as independent or synergistic determinants of the selected outcomes. Spanning from birth defects to mortality, these outcomes include: chromosomal disorders, infectious and non-infectious disease, occupational and environmental disease, trauma, pregnancy, menopause, and access to health services. As these examples highlight, not only can gender relations influence expression-and interpretation-of biological traits, but also sex-linked biological characteristics can, in some cases, contribute to or amplify gender differentials in health. Because our science will only be as clear and error-free as our thinking, greater precision about whether and when gender relations, sex-linked biology, both, or neither matter for health is warranted. PMID- 12913048 TI - Epidemiology on the side of the angels...Or the people? PMID- 12913049 TI - Biological warfare and the people of Iraq. PMID- 12913050 TI - Global trends in adenocarcinomas and obesity: an epidemiologic link? PMID- 12913054 TI - Effects of heart rate changes on arterial distensibility in humans. AB - In rats, an increase in heart rate by pacing is accompanied by progressive large artery stiffening. Whether this is also the case in humans is unknown. We enrolled 20 patients who were chronically implanted with a pacemaker because of atrioventricular block or sick sinus syndrome. Arterial distensibility was measured by an echo-tracking device. In 10 patients, the evaluation was performed on the radial artery by using continuous finger blood pressure measurements, whereas in the remaining 10 patients, the common carotid artery was studied with a semiautomatic measure of brachial artery blood pressure. Diastolic diameter, systodiastolic diameter change, and distensibility were obtained at baseline (heart rate 63+/-2 beats/min) and after atrial and ventricular sequential pacing at a heart rate of 90 and 110 beats/min. At baseline, the diameter was 7.8+/-0.3 mm in the carotid artery and 2.4+/-0.1 mm in the radial artery; the respective systodiastolic diameter change values were 375.4+/-31.0 and 55.9+/-9.0 (microm) and the distensibility values were 1.4+/-0.1 and 0.7+/-0.1 (1/mm Hg 10-3). Blood pressure and diameter were not significantly modified by increasing heart rate, which markedly modified systodiastolic diameter change and distensibility. In the radial artery, distensibility was reduced by 47% (P<0.05) at a heart rate of 90 beats/min with no further reduction at 110 beats/min. In the carotid artery, distensibility was reduced by 20% at a heart rate of 90 beats/min (P<0.05) with a further reduction at 110 beats/min (45%, P<0.05). These data provide the first evidence in humans that acute increases in heart rate markedly affect arterial distensibility and that this occurs in both large- and middle-size muscle arteries within the range of "normal" heart rate values. PMID- 12913055 TI - Hormone replacement therapy and inflammation: interactions in cardiovascular disease. AB - Inflammation plays a central role in the pathogenesis of many forms of vascular disease, including atherosclerosis. Atherogenesis begins with endothelial damage, and the damaged endothelium expresses adhesion molecules, chemokines, and proinflammatory cytokines that direct atherosclerotic plaque formation and spill into the circulation as biomarkers of atherosclerotic disease risk. Menopausal hormone therapy, including a variety of estrogen preparations with or without a progestin, has negative modulatory effects on most of these soluble inflammatory markers, including E-selectin, vascular cell adhesion molecule-1, intercellular adhesion molecule-1, monocyte chemoattractant protein-1, and tumor necrosis factor-alpha, inconsistent effects on interleukin-6, and stimulatory effects on transforming growth factor-beta, a vasoprotective cytokine. In contrast, C reactive protein, a circulating proinflammatory cytokine produced in both liver and atherosclerotic arteries, increases in response to oral conjugated estrogens but not to transdermal estrogen. Although C-reactive protein is clearly linked to increased cardiovascular disease risk in women, the hormone-induced rise in this biomarker is not associated with increased risk and may be related to a first pass effect of C-reactive protein production in the liver after oral estrogen absorption. Many important questions about the effects of ovarian hormones on vascular inflammation and the pathogenesis of vascular disease cannot be answered in human subjects. Insights from fundamental mechanistic studies in animal models are needed to delineate the cellular/molecular events that determine whether these hormones protect or injure blood vessels. PMID- 12913056 TI - Gene transfer of eNOS to the thick ascending limb of eNOS-KO mice restores the effects of L-arginine on NaCl absorption. AB - The thick ascending limb of the loop of Henle (THAL) plays an essential role in the regulation of sodium and water homeostasis by the kidney. l-Arginine, the substrate for nitric oxide synthase (NOS), decreases NaCl absorption by THALs. We hypothesized that eNOS produces the NO that regulates THAL NaCl transport and that selective expression of eNOS in the THAL of eNOS knockout(-/-) mice would restore the effects of l-arginine on NaCl absorption. eNOS-/- mice were anesthetized, the left kidney was exposed, and the renal interstitium was injected with recombinant adenoviral vectors that expressed green fluorescent protein (GFP) or eNOS driven by the promoter of the Na/K/2Cl cotransporter Ad NKCC2GFP and Ad-NKCC2eNOS, respectively. In Ad-NKCC2eNOS-transduced kidneys, eNOS expression was detected 7 days after injection but was absent in Ad-NKCC2GFP transduced kidneys. In THALs from eNOS-/- mice transduced with Ad-NKCC2eNOS, adding L-arginine increased DAF-2DA fluorescence, a measure of NO production, by 9.1+/-1.1% (P<0.05; n=5), but not in THALs transduced with Ad-NKCC2GFP. In THALs from eNOS-/- mice transduced with Ad-NKCC2eNOS, Cl absorption averaged 85.9+/ 11.8 pmol/min per millimeter. Adding l-arginine (1 mmol/L) to the bath decreased Cl absorption to 59.7+/-11.0 pmol/min per millimeter (P<0.05; n=6). In THALs transduced with Ad-NKCC2GFP, Cl absorption averaged 96.0+/-21.0 pmol/min per millimeter. Adding L-arginine to the bath did not significantly affect Cl absorption (100.6+/-20.6 pmol/min per millimeter; n=4). We concluded that gene transfer of eNOS to the THAL of eNOS-/- mice restores L-arginine-induced inhibition of NaCl transport and NO production. These data indicate that eNOS is essential for the regulation of THAL NaCl transport by NO. PMID- 12913057 TI - Training-induced, pressure-lowering effect in SHR: wide effects on circulatory profile of exercised and nonexercised muscles. AB - We showed that the training-induced, pressure-lowering effect correlates with decreased arteriole wall/lumen ratio and venule growth in the gracilis muscle. To investigate whether these beneficial changes are tissue-specific or occur in other muscles and tissues, we analyzed the effects of hypertension and training on microcirculatory profile of locomotor/nonlocomotor muscles and another nonmuscular tissue. Spontaneously hypertensive rats (SHR) and Wistar-Kyoto rats were submitted to low-intensity training (50% to 60% maximal exercise capacity, 13 weeks); age-matched control rats were kept sedentary. Trained and sedentary rats were instrumented for pressure and heart rate measurements at rest. Morphometric analyses (arterioles, capillaries, venules) were performed in all tissues. Training attenuated pressure and heart rate only in SHR. Arterioles (inner diameter <30 microm) were markedly hypertrophied in sedentary SHR, but wall/lumen ratio was equally reduced (approximately 30%) and normalized by training in locomotor (soleus, gastrocnemius, gracilis) and nonlocomotor skeletal muscles (temporalis) in the myocardium and diaphragm, without changes in the renal cortex. Training also increased venule density (approximately 2-fold) only in locomotor and nonlocomotor muscles of SHR. Capillary density was similarly increased in all exercised muscles of both groups, with no change in temporalis and kidneys. Data suggest that growth/proliferation of small venules and regression of hypertrophied arteriole wall/lumen ratio are generalized tissue specific (skeletal muscle) and group-specific (SHR) adjustments to training to reduce local resistance and augment physical capacity of circulation, thus contributing to training-induced pressure-lowering effect. They are accompanied by remodeling of myocardium (cardiac output) and diaphragm arterioles (ventilatory adjustments), stressing the importance of training as a nonpharmacological therapy to control pressure levels in hypertension. PMID- 12913058 TI - Role of endogenous angiotensin II on glutamatergic actions in the rostral ventrolateral medulla in Goldblatt hypertensive rats. AB - In this study, we investigated the cardiovascular responses mediated by rostral ventrolateral medulla neurons (RVLM) in the Goldblatt hypertension model (2K-1C) treated or not treated with captopril. The actions of glutamate into the RVLM were tested, injecting glutamate (0.1 mol/L, 100 nL) and its antagonist kynurenic acid (0.02 mol/L, 100 nL). Glycine (0.5 mol/L, 100 nL) was also microinjected. Experiments were performed in male Wistar rats (weight, 250 to 300 g); 5 groups were studied: (1) 2K-1C nontreated (H, n=6); (2) 2K-1C treated with captopril, 10 mg/kg per day (Ht10, n=10); (3) 2K-1C treated with captopril, 50 mg/kg per day (Ht50, n=7); (4) control normotensive rats (N, n=7); and (5) normotensive rats treated with captopril, 50 mg/kg per day (Nt50, n=8). All experiments in 2K-1C were performed 6 weeks after renal surgery; captopril treatment lasted for the last 2 weeks. In urethane-anesthetized rats (1.2 g/kg IV), bilateral microinjection of glycine into the RVLM caused a depressor response; there was no difference between groups in relation to the change of variation (N: 54+/-2; H: 46+/-12; Ht10: 50+/-3, and Ht50: 42+/-7 mm Hg). Only in the H group, kynurenic acid microinjection into the RVLM caused a depressor response (H: 158+/-8 to 132+/-8 mm Hg). Glutamate response was larger in hypertensive than in normotensive rats (N: 38+/-2.6 and H: 55+/-6); no difference was observed between hypertensive groups. The data suggest that glutamate acts tonically to drive the RVLM in 2K-1C rats, and this action is modulated by endogenous angiotensin II. The increase in the glutamate actions within the RVLM may contribute to the pathogenesis of renovascular hypertension. PMID- 12913059 TI - African Americans with LVH demonstrate depressed sensitivity of the coronary microcirculation to stimulated relaxation. AB - Excess coronary heart disease morbidity and mortality among African Americans remains an important yet unexplained public health problem. We hypothesized that adverse outcome is in part due to intrinsic or acquired abnormalities in coronary endothelial function and vasoreactivity. We compared dose-response curves relating changes in coronary blood flow and epicardial diameter to graded infusions of acetylcholine in 50 African American and 65 white subjects with hypertensive left ventricular hypertrophy (LVH) and normal coronary arteries. These groups were similar for age, body mass index, mean arterial pressure, and indexed left ventricular mass. The same protocol was conducted in 24 normotensive African American and 56 similar white subjects. We found significant depression in the coronary blood flow dose-response curve relation among African Americans when compared with white subjects with similar LVH (P<0.03). Racial differences were observed at all doses of acetylcholine but were less precisely estimated at the highest dose. The same testing among normotensive subjects revealed similar dose-response curves with no significant effect of race. Qualitatively similar results were found with respect to coronary diameter. Adenosine responses, a measure of endothelium-independent function, were similar after partitioning by LVH. Our study demonstrates that there are racial differences in sensitivity of coronary arteries to acetylcholine-stimulated relaxation among those with LVH. These results provide a mechanism whereby racial differences in coronary vasoreactivity might contribute to adverse coronary heart disease outcome among African Americans, a group in whom LVH is prevalent. PMID- 12913060 TI - Coronary angiography is the best predictor of events in renal transplant candidates compared with noninvasive testing. AB - Guidelines for the detection of coronary artery disease (CAD) and assess of risk in renal transplant candidates are based on the results of noninvasive testing, according to data originated in the nonuremic population. We evaluated prospectively the accuracy of 2 noninvasive tests and risk stratification in detecting CAD (>or=70% obstruction) and assessing cardiac risk by using coronary angiography (CA). One hundred twenty-six renal transplant candidates who were classified as at moderate (>or=50 years) or high (diabetes, extracardiac atherosclerosis, or clinical coronary artery disease) coronary risk underwent myocardial scintigraphy (SPECT), dobutamine stress echocardiography, and CA and were followed for 6 to 48 months. The prevalence of CAD was 42%. The sensitivities and negative predictive values for the 2 noninvasive tests and risk stratification were <75%. After 6 to 48 months, there were 18 cardiac events, 9 fatal. Risk stratification (P=0.007) and CA (P=0.0002) predicted the crude probability of surviving free of cardiac events. The probability of event-free survival at 6, 12, 24, 36, and 48 months were 98%, 98%, 94%, 94%, and 94% in patients with <70% stenosis on CA and 97%, 87%, 61%, 56%, and 54% in patients with >or=70% stenosis. Multivariate analysis showed that the sole predictor of cardiac events was critical coronary lesions (P=0.003). Coronary angiography may still be necessary for detecting CAD and determining cardiac risk in renal transplant candidates. The data suggest that current algorithms based on noninvasive testing in this population should be revised. PMID- 12913061 TI - Catecholamines block the antimitogenic effect of estradiol on human glomerular mesangial cells. AB - Local sequential conversion of estradiol to hydroxyestradiols and methoxyestradiols by CYP450 and catechol-O-methyltransferase, respectively, contributes to the antimitogenic effects of estradiol on glomerular mesangial cell growth via estrogen receptor-independent mechanisms. Catecholamines are also substrates for catechol-O-methyltransferase and therefore, might abrogate the renoprotective effects of estradiol by inhibiting formation of methoxyestradiols. To test this hypothesis, we investigated the antimitogenic effects of estradiol on human glomerular mesangial cell proliferation and collagen synthesis in the presence and absence of catecholamines. Norepinephrine, epinephrine, and isoproterenol abrogated the inhibitory effects of estradiol on cell number, DNA synthesis, and collagen synthesis. For example, serum-induced DNA synthesis was inhibited from 100% to 62+/-1.9% by 0.1 micromol/L estradiol, and these inhibitory effects were reversed to 91+/-1.9% by 1 micromol/L epinephrine, 90.7+/ 3.3% by 1 micromol/L isoproterenol, 87.5+/-2.8% by 10 micromol/L norepinephrine, and 92+/-1% by 10 micromol/L OR486 (catechol-O-methyltransferase inhibitor). The interaction of catecholamines with estradiol was not affected by phentolamine or propanolol, alpha- and beta-adrenoceptor antagonists, respectively. Similar to estradiol, the antimitogenic effects of 2-hydroxyestradiol were abrogated by epinephrine, isoproterenol, and OR486. In contrast to estradiol and 2 hydroxyestradiol, the antimitogenic effects of 2-methoxyestradiol were not attenuated by epinephrine, isoproterenol, or OR486. Norepinephrine, epinephrine, and isoproterenol inhibited the conversion of both estradiol and 2 hydroxyestradiol to 2-methoxyestradiol. Our findings suggest that catecholamines within the glomeruli might abrogate the antimitogenic effects of estradiol by blocking the conversion of 2-hydroxyestradiol to 2-methoxyestradiol. PMID- 12913062 TI - SR141716A-sensitive enhancement of ET-1 hypotensive effect by chronic NOS inhibition. AB - The present study evaluated the potential mechanism involved in the hypotensive effect induced by ET-1 in rats treated with the NO synthase inhibitor NG-nitro-L arginine methyl ester (L-NAME) in the drinking water during 7 days. Hypertension developed in the L-NAME-treated rats (164+/-3 versus 112+/-1 mm Hg in untreated control rats), and the hypotensive effect of ET-1 (100 pmol/kg IV) was significantly enhanced compared with control rats (32+/-2% versus 20+/-1% fall in mean arterial pressure). The enhanced ET-1 hypotensive effect in L-NAME-treated rats was abolished by the ETB receptor antagonist BQ-788 but was unaltered by the cyclooxygenase inhibitor diclofenac, the cytochrome P450 inhibitor fluconazole, or the potassium channel blockers apamin, glibenclamide, tetraethylammonium, and 4-aminopyridine. Pretreatment with the cannabinoid CB1 receptor antagonist SR141716A significantly reduced the hypotensive response to ET-1 in L-NAME treated rats (20+/-1%), although it did not modify the response in untreated control rats (17+/-1%). These findings indicate that in rats under chronic NOS inhibition, the hypotensive effect of ET-1 is unexpectedly enhanced and appears to be mediated by a non-NO/non-prostanoid mechanism and involves an SR141716A sensitive mechanism triggered by ETB receptor activation. PMID- 12913063 TI - ETA receptor blockade decreases vascular superoxide generation in DOCA-salt hypertension. AB - Development and progression of end-organ damage in hypertension have been associated with increased oxidative stress. Superoxide anion accumulation has been reported in deoxycorticosterone acetate (DOCA)-salt hypertension, in which endothelin-1 plays an important role in cardiovascular damage. We hypothesized that blockade of ETA receptors in DOCA-salt rats would decrease oxidative stress. Both systolic blood pressure (SBP, 210+/-9 mm Hg; P<0.05) and vascular superoxide generation in vivo were increased in DOCA-salt (44.9+/-10.3% of ethidium bromide positive nuclei; P<0.05) versus control uninephrectomized (UniNx) rats (118+/-3 mm Hg; 18.5+/-3%, respectively). In DOCA-salt rats, the ETA antagonist BMS 182874 (40 mg/kg per day PO) lowered SBP (170+/-4 versus UniNx, 120+/-3 mm Hg) and normalized superoxide production (21.7+/-6 versus UniNx, 11.9+/-7%). Vitamin E (200 mg/kg per day PO) decreased superoxide formation in DOCA-salt rats (18.8+/ 7%) but did not alter SBP. Oxidative stress in nonstimulated circulating polymorphonuclear cells (PMNs) or in PMNs treated with zymosan, an inducer of superoxide release, was similar in DOCA-salt and UniNx groups. Superoxide formation by PMNs was unaffected by treatment with BMS 182874. Western blot analysis showed increased nitrotyrosine-containing proteins in mesenteric vessels from DOCA-salt compared with UniNX. Treatment with either BMS 182874 or vitamin E abolished the differences in vascular nitrotyrosine-containing proteins between DOCA-salt and UniNX. Maximal relaxation to acetylcholine was decreased in DOCA salt aortas (75.8+/-4.2% versus UniNx, 95.4+/-1.9%, P<0.05). BMS 182874 treatment increased acetylcholine-induced relaxation in DOCA-salt aortas to 93.5+/-4.5%. These in vivo findings indicate that increased vascular superoxide production is associated with activation of the endothelin system through ETA receptors in DOCA salt hypertension, in apparently blood pressure-independent fashion. PMID- 12913064 TI - Chronic salt loading upregulates expression of adrenomedullin and its receptors in adrenal glands and kidneys of the rat. AB - The vasodilator peptide adrenomedullin (AM) elicits diuresis and natriuresis and inhibits aldosterone secretion. The aim of this study was to better understand the role of AM in maintaining water and electrolyte balance during chronic salt loading. Male Wistar rats were divided into a high salt (HS) group that received a diet containing 8% sodium chloride (NaCl) and a normal salt group that received a diet containing 0.4% NaCl. Plasma AM concentrations as well as expression of AM mRNA in the adrenal gland and kidney were then measured after 3, 7, 14, and 28 days. After 28 days, sodium and water excretion were significantly higher in HS rats than in control, although blood pressure and fluid volume were not significantly affected. Moreover, although plasma AM remained unchanged for up to 14 days, it was increased 2.5-fold in HS rats after 28 days on a high salt diet, and there were corresponding 3-fold and 1.5-fold increases in the levels of AM mRNA in the adrenal gland and kidney, respectively. At the same time, expression of calcitonin receptor-like receptor mRNA was significantly upregulated in both kidney and adrenal gland, as was expression of receptor activity-modify protein 1 (RAMP1) and RAMP2 mRNA in the adrenals and expression of RAMP3 in kidneys. Taken together, these results suggest that AM plays a role in the regulation of water and electrolyte balance in animals chronically ingesting high levels of salt. PMID- 12913065 TI - Interaction of carbon monoxide and adenosine in the nucleus tractus solitarii of rats. AB - Carbon monoxide has been identified as an endogenous biological messenger in the brain. Heme oxygenase catalyzes the metabolism of heme to carbon monoxide and biliverdin. Previously, we have shown the involvement of carbon monoxide in central cardiovascular regulation, baroreflex modulation, and glutamatergic neurotransmission in the nucleus tractus solitarii of rats. We also showed that adenosine increased the release of glutamate in the nucleus tractus solitarii. In this study, we investigated the possible interactions of carbon monoxide and adenosine in the nucleus tractus solitarii. Male Sprague-Dawley rats were anesthetized with urethane, and blood pressure were monitored intra-arterially. Unilateral microinjection of increasing doses of hemin (0.01 to 3.3 nmol), a heme molecule cleaved by heme oxygenase to yield carbon monoxide, produced a significant decrease in blood pressure and heart rate in a dose-dependent manner. In addition, similar cardiovascular effects were observed after injection of adenosine (2.3 nmol). These cardiovascular effects of hemin were attenuated by prior administration of the adenosine receptor antagonist 1,3-dipropyl-8 sulfophenylxanthine. Similarly, pretreatment of the heme oxygenase inhibitor zinc protoporphyrin IX or zinc deuteroporphyrin 2,4-bis glycol also attenuated the depressor and bradycardic effects of adenosine. These results indicate that the interaction between carbon monoxide and adenosine may contribute to the activation of heme oxygenase in central cardiovascular regulation. PMID- 12913066 TI - Ubiquitin carboxy-terminal hydrolase L1 binds to and stabilizes monoubiquitin in neuron. AB - Mammalian neuronal cells abundantly express a deubiquitylating enzyme, ubiquitin carboxy-terminal hydrolase 1 (UCH L1). Mutations in UCH L1 are linked to Parkinson's disease as well as gracile axonal dystrophy (gad) in mice. In contrast to the UCH L3 isozyme that is universally expressed in all tissues, UCH L1 is expressed exclusively in neurons and testis/ovary. We found that UCH L1 associates and colocalizes with monoubiquitin and elongates ubiquitin half-life. The gad mouse, in which the function of UCH L1 is lost, exhibited a reduced level of monoubiquitin in neurons. In contrast, overexpression of UCH L1 caused an increase in the level of ubiquitin in both cultured cells and mice. These data suggest that UCH L1, with avidity and affinity for ubiquitin, insures ubiquitin stability within neurons. This study is the first to show the function of UCH L1 in vivo. PMID- 12913067 TI - A genomic rearrangement resulting in a tandem duplication is associated with split hand-split foot malformation 3 (SHFM3) at 10q24. AB - Split hand-split foot malformation (SHFM) is characterized by hypoplasia/aplasia of the central digits with fusion of the remaining digits. SHFM is usually an autosomal dominant condition and at least five loci have been identified in humans. Mutation analysis of the DACTYLIN gene, suspected to be responsible for SHFM3 in chromosome 10q24, was conducted in seven SHFM patients. We screened the coding region of DACTYLIN by single-strand conformation polymorphism and sequencing, and found no point mutations. However, Southern, pulsed field gel electrophoresis and dosage analyses demonstrated a complex rearrangement associated with a approximately 0.5 Mb tandem duplication in all the patients. The distal and proximal breakpoints were within an 80 and 130 kb region, respectively. This duplicated region contained a disrupted extra copy of the DACTYLIN gene and the entire LBX1 and beta-TRCP genes, known to be involved in limb development. The possible role of these genes in the SHFM3 phenotype is discussed. PMID- 12913068 TI - Chromosome 12q harbors multiple genetic loci related to asthma and asthma-related phenotypes. AB - Chromosome 12q13-24 is among the regions most frequently identified in genome wide surveys for asthma susceptibility loci, with reports of two distinct clusters of positive linkage signals: one near the interferon gamma locus, the other near the nitric oxide synthase 1 locus. These results suggest that 12q harbors several asthma susceptibility loci. We evaluated this possibility in a subset of families ascertained through the Childhood Asthma Management Program (CAMP) Genetics Ancillary Study. Fifty-five nuclear families with at least two asthmatic siblings (212 individuals) were genotyped using 32 microsatellite markers. Non-parametric linkage analysis was performed for the asthma phenotype (qualitative). Multipoint variance component-based linkage analysis was performed for five quantitative asthma-related traits: (i) percent predicted forced expiratory volume in one second (FEV(1)); (ii) dose of methacholine resulting in 20% fall in FEV(1) from baseline (PC(20)); (iii) post-bronchodilator percent change in FEV(1) (BDPR); (iv) serum eosinophil levels (EOS); and (v) total serum IgE levels (IgE). Three separate and distinct loci demonstrated evidence suggestive of linkage: asthma at 68 cM (exact P-value=0.05), airways responsiveness (PC(20)) at 147 cM (P=0.01), and indices of pulmonary function (FEV1, BDPR) at 134 cM (P=0.05 and P<0.01, respectively). No linkage was observed for the atopy-related phenotypes. We provide further evidence supporting the presence of an asthma susceptibility locus at the proximal end of chromosome 12q, as well as new evidence for additional loci more distally that account for unique features of the asthma phenotype. Fine mapping efforts for these loci are warranted. PMID- 12913069 TI - Fgd1, the Cdc42 GEF responsible for Faciogenital Dysplasia, directly interacts with cortactin and mAbp1 to modulate cell shape. AB - FGD1 mutations result in Faciogenital Dysplasia (FGDY), an X-linked human disease that affects skeletal formation and embryonic morphogenesis. FGD1 and Fgd1, the mouse FGD1 ortholog, encode guanine nucleotide exchange factors (GEF) that specifically activate Cdc42, a Rho GTPase that controls the organization of the actin cytoskeleton. To further understand FGD1/Fgd1 signaling and begin to elucidate the molecular pathophysiology of FGDY, we demonstrate that Fgd1 directly interacts with cortactin and mouse actin-binding protein 1 (mAbp1), actin-binding proteins that regulate actin polymerization through the Arp2/3 complex. In yeast two-hybrid studies, cortactin and mAbp1 Src homology 3 (SH3) domains interact with a single Fgd1 SH3-binding domain (SH3-BD), and biochemical studies show that the Fgd1 SH3-BD directly binds to cortactin and mAbp1 in vitro. Immunoprecipitation studies show that Fgd1 interacts with cortactin and mAbp1 in vivo and that Fgd1 SH3-BD mutations disrupt binding. Immunocytochemical studies show that Fgd1 colocalizes with cortactin and mAbp1 in lamellipodia and membrane ruffles, and that Fgd1 subcellular targeting is dynamic. By using truncated cortactin proteins, immunocytochemical studies show that the cortactin SH3 domain targets Fgd1 to the subcortical actin cytoskeleton, and that abnormal Fgd1 localization results in actin cytoskeletal abnormalities and significant changes in cell shape and viability. Thus, this study provides novel in vitro and in vivo evidence that Fgd1 specifically and directly interacts with cortactin and mAbp1, and that these interactions play an important role in regulating the actin cytoskeleton and, subsequently, cell shape. PMID- 12913070 TI - Zinc metalloproteinase, ZMPSTE24, is mutated in mandibuloacral dysplasia. AB - Mandibuloacral dysplasia (MAD; OMIM 248370) is a rare, genetically and phenotypically heterogeneous, autosomal recessive disorder characterized by skeletal abnormalities including hypoplasia of the mandible and clavicles, acro osteolysis, cutaneous atrophy and lipodystrophy. A homozygous missense mutation, Arg527His, in the LMNA gene which encodes nuclear lamina proteins lamins A and C has been reported in patients with MAD and partial lipodystrophy. We studied four patients with MAD who had no mutations in the LMNA gene. We now show compound heterozygous mutations, Phe361fsX379 and Trp340Arg, in the zinc metalloproteinase (ZMPSTE24) gene in one of the four patients who had severe MAD associated with progeroid appearance and generalized lipodystrophy. ZMPSTE24 is involved in post translational proteolytic cleavage of carboxy terminal residues of farnesylated prelamin A in two steps to form mature lamin A. Deficiency of Zmpste24 in mice causes accumulation of prelamin A and phenotypic features similar to MAD. The yeast homolog, Ste24, has a parallel role in processing of prenylated mating pheromone a-factor. Since human ZMPSTE24 can also process a-factor when expressed in yeast, we assessed the functional significance of the two ZMPSTE24 mutations in the yeast to complement the mating defect of the haploid MATa yeast lacking STE24 and Ras-converting enzyme 1 (RCE1; another prenylprotein-specific endoprotease) genes. The ZMPSTE24 mutant construct, Phe361fsX379, was inactive in complementing the yeast a-factor but the mutant, Trp340Arg, was partially active compared to the wild type ZMPSTE24 construct. We conclude that mutations in ZMPSTE24 may cause MAD by affecting prelamin A processing. PMID- 12913071 TI - Wolfram syndrome: structural and functional analyses of mutant and wild-type wolframin, the WFS1 gene product. AB - Mutations of the WFS1 gene are responsible for Wolfram syndrome, a rare, recessive disorder characterized by early-onset, non-autoimmune diabetes mellitus, optic atrophy and further neurological and endocrinological abnormalities. The WFS1 gene encodes wolframin, a putative multispanning membrane glycoprotein of the endoplasmic reticulum. The function of wolframin is completely unknown. In order to characterize wolframin, we have generated polyclonal antibodies against both hydrophilic termini of the protein. Wolframin was found to be ubiquitously expressed with highest levels in brain, pancreas, heart and insulinoma beta-cell lines. Analysis of the structural features provides experimental evidence that wolframin contains nine transmembrane segments and is embedded in the membrane in an N(cyt)/C(lum) topology. Wolframin assembles into higher molecular weight complexes of approximately 400 kDa in the membrane. Pulse-chase experiments demonstrate that during maturation wolframin is N-glycosylated but lacks proteolytical processing. Moreover, N-glycosylation appears to be essential for the biogenesis and stability of wolframin. Here we investigate, for the first time, the molecular mechanisms that cause loss-of function of wolframin in affected individuals. In patients harboring nonsense mutations complete absence of the mutated wolframin is caused by instability and rapid decay of WFS1 nonsense transcripts. In a patient carrying a compound heterozygous missense mutation, R629W, we found markedly reduced steady-state levels of wolframin. Pulse-chase experiments of mutant wolframin expressed in COS 7 cells indicated that the R629W mutation leads to instability and strongly reduced half-life of wolframin. Thus, the Wolfram syndrome in patients investigated here is caused by reduced protein dosage rather than dysfunction of the mutant wolframin. PMID- 12913072 TI - Global disruption of the cerebellar transcriptome in a Down syndrome mouse model. AB - Trisomy 21 (Down syndrome) results in cerebellar dysmorphology with direct parallels in the Ts65Dn mouse. Despite pronounced changes in morphology, cerebellar function is not markedly different. As a first test of whether those cerebellar cells that have survived to adulthood in trisomic mice are equivalent to euploid cells, we used microarrays to assess the trisomic and euploid cerebella. Trisomic and euploid transcriptomes were robustly distinguished. Changes in expression of individual genes were very subtle, but the differences in respective transcriptome phenotypes extended deeply into the set of nearly 7000 probes (genes) located throughout the genome. In contrast to deterministic models of gene action in trisomy, examination of the discriminating genes in two independent experiments suggests that the global perturbation includes a significant stochastic component. Thus, dosage imbalance of 124 genes in Ts65Dn mice alters the expression of thousands of genes to create a variable trisomic transcriptome. This global destabilization has important implications for approaches to ameliorative therapies in Down syndrome. PMID- 12913073 TI - Abnormal association of mutant huntingtin with synaptic vesicles inhibits glutamate release. AB - In Huntington disease (HD), polyglutamine expansion causes the disease protein huntingtin to aggregate and accumulate in the nucleus and cytoplasm. The cytoplasmic huntingtin aggregates are found in axonal terminals and electrophysiological studies show that mutant huntingtin affects synaptic neurotransmission. However, the biochemical basis for huntingtin-mediated synaptic dysfunction is unclear. Using electron microscopy on sections of HD mouse brains, we found that axonal terminals containing huntingtin aggregates often had fewer synaptic vesicles than did normal axonal terminals. Subcellular fractionation and electron microscopy revealed that mutant huntingtin is co localized with huntingtin-associated protein-1 (HAP1) in axonal terminals in the brains of HD transgenic mice. Mutant huntingtin binds more tightly to synaptic vesicles than does normal huntingtin, and it decreases the association of HAP1 with synaptic vesicles in HD mouse brains. Brain slices from HD transgenic mice that had axonal aggregates showed a significant decrease in [(3)H]glutamate release, suggesting that neurotransmitter release from synaptic vesicles was impaired. Taken together, these findings suggest that mutant huntingtin has an abnormal association with synaptic vesicles and this association impairs synaptic function. PMID- 12913074 TI - Characterization of disease-associated mutations affecting an exonic splicing enhancer and two cryptic splice sites in exon 13 of the cystic fibrosis transmembrane conductance regulator gene. AB - Sequences in exons can play an important role in constitutive and regulated pre mRNA splicing. Since exonic splicing regulatory sequences are generally poorly conserved and their mechanism of action is not well understood, the consequence of exonic mutations on splicing can only be determined empirically. In this study, we have investigated the consequence of two cystic fibrosis (CF) disease causing mutations, E656X and 2108delA, on the function of a putative exonic splicing enhancer (ESE) in exon 13 of the CFTR gene. We have also determined whether five other CF mutations D648V, D651N, G654S, E664X and T665S located near this putative ESE could lead to aberrant splicing of exon 13. Using minigene constructs, we have demonstrated that the E656X and 2108delA mutations could indeed cause aberrant splicing in a predicted manner, supporting a role for the putative ESE sequence in pre-mRNA splicing. In addition, we have shown that D648V, E664X and T665S mutations could cause aberrant splicing of exon 13 by improving the polypyrimidine tracts of two cryptic 3' splice sites. We also provide evidence that the relative levels of two splicing factors, hTra2alpha and SF2/ASF, could alter the effect on splicing of some of the exon 13 disease mutations. Taken together, our results suggest that the severity of CF disease could be modulated by changes in the fidelity of CFTR pre-mRNA splicing. PMID- 12913075 TI - TBX1 is required for inner ear morphogenesis. AB - TBX1 is thought to be a critical gene in the pathogenesis of del22q11/DiGeorge syndrome (DGS). Morphological abnormalities of the external ear and hearing impairment (conductive or sensorineural) affect the majority of patients. Here we show that homozygous mutation of the mouse homolog Tbx1 is associated with severe inner ear defects that prevent the formation of the cochlea and of the vestibulum. Consistent with phenotypic abnormalities, Tbx1 is expressed early in otocyst development in the otic epithelium and in the periotic mesenchyme. Tbx1 loss-of-function blocks inner ear development at early otocyst stage and after neurogenesis. Analysis of chimeras suggests that Tbx1 function is required in the otic epithelium cell autonomously, but abnormalities of the periotic mesenchyme indicate that the pathogenesis of the inner ear phenotype is complex. We propose a model where Tbx1 is required for expansion of a subpopulation of otic epithelial cells, which is required to form the vestibular and auditory organs. Our data suggest that Tbx1 deletion in del22q11 patients may cause not only external and middle ear defects but also sensorineural and vestibular phenotypes observed in these patients. PMID- 12913076 TI - Claudin 14 knockout mice, a model for autosomal recessive deafness DFNB29, are deaf due to cochlear hair cell degeneration. AB - Tight junctions (TJs) create ion-selective paracellular permeability barriers between extracellular compartments. In the organ of Corti of the inner ear, TJs of the reticular lamina separate K(+)-rich endolymph and Na(+)-rich perilymph. In humans, mutations of the gene encoding claudin 14 TJ protein cause profound deafness but the underlying pathogenesis is unknown. To explore the role of claudin 14 in the inner ear and in other tissues we created a mouse model by a targeted deletion of Cldn14. In the targeted allele a lacZ cassette is expressed under the Cldn14 promoter. In Cldn14-lacZ heterozygous mice beta-galactosidase activity was detected in cochlear inner and outer hair cells and supporting cells, in the collecting ducts of the kidney, and around the lobules of the liver. Cldn14-null mice have a normal endocochlear potential but are deaf due to rapid degeneration of cochlear outer hair cells, followed by slower degeneration of the inner hair cells, during the first 3 weeks of life. Monolayers of MDCK cells expressing claudin 14 show a 6-fold increase in the transepithelial electrical resistance by decreasing paracellular permeability for cations. In wild type mice, claudin 14 was immunolocalized at hair cell and supporting cell TJs. Our data suggest that the TJ complex at the apex of the reticular lamina requires claudin 14 as a cation-restrictive barrier to maintain the proper ionic composition of the fluid surrounding the basolateral surface of outer hair cells. PMID- 12913077 TI - Targeted disruption of exons 1 to 6 of the Fanconi Anemia group A gene leads to growth retardation, strain-specific microphthalmia, meiotic defects and primordial germ cell hypoplasia. AB - Fanconi Anemia (FA) is an autosomal recessive disorder characterized by cellular hypersensitivity to DNA cross-linking agents. Recent studies suggest that FA proteins share a common pathway with BRCA proteins. To study the in vivo role of the FA group A gene (Fanca), gene-targeting techniques were used to generate Fanca(tm1Hsc) mice in which Fanca exons 1-6 were replaced by a beta-galactosidase reporter construct. Fanca(tm1.1Hsc) mice were generated by Cre-mediated removal of the neomycin cassette in Fanca(tm1Hsc) mice. Fanca(tm1.1Hsc) homozygotes display FA-like phenotypes including growth retardation, microphthalmia and craniofacial malformations that are not found in other Fanca mouse models, and the genetic background affects manifestation of certain phenotypes. Both male and female mice homozygous for Fanca mutation exhibit hypogonadism, and homozygous females demonstrate premature reproductive senescence and an increased incidence of ovarian cysts. We showed that fertility defects in Fanca(tm1.1Hsc) homozygotes might be related to a diminished population of primordial germ cells (PGCs) during migration into the gonadal ridges. We also found a high level of Fanca expression in pachytene spermatocytes. Fanca(tm1Hsc) homozygous males exhibited an elevated frequency of mispaired meiotic chromosomes and increased apoptosis in germ cells, implicating a role for Fanca in meiotic recombination. However, the localization of Rad51, Brca1, Fancd2 and Mlh1 appeared normal on Fanca(tm1Hsc) homozygous meiotic chromosomes. Taken together, our results suggest that the FA pathway plays a role in the maintenance of reproductive germ cells and in meiotic recombination. PMID- 12913078 TI - A genome wide scan for early onset primary hypertension in Scandinavians. AB - With the aim of identifying hypertension susceptibility loci, we performed a genome wide scan in Scandinavian sib-pairs with early onset primary hypertension. To be classified as affected, a diagnosis of primary hypertension at age /= 1.0) were fine mapped with additional markers. Multipoint non-parametric linkage analysis was performed using GENEHUNTER v 2.0. Using simulations, a nominal P5 years. CONCLUSIONS: In most cases, cHCC-CC seems to be a variant of ordinary HCC with cholangiocellular features, rather than a true intermediate disease entity between HCC and CC. The surgical approach is recommended for selected patients with cHCC-CC. PMID- 12913083 TI - Results of a randomized trial with or without 5-FU-based preoperative chemotherapy followed by postoperative chemotherapy in resected colon and rectal carcinoma. AB - BACKGROUND: Our previous study confirmed the efficacy of postoperative treatment with mitomycin C (MMC) and oral 5-fluorouracil (5-FU) for colorectal cancer. The 2nd trial was designed to evaluate the effectiveness of additional preoperative chemotherapy to postoperative treatment with MMC and oral 5-FU for curatively resected colorectal cancer patients. PATIENTS AND METHODS: 1355 patients (colon 755; rectum 600) were enrolled in this study. The pre- and postoperative chemotherapy (PPC) group was treated preoperatively with 5-FU (320 mg/m(2)/day) by continuous intravenous infusion for 5 days beginning on day 6 before surgery and postoperatively with MMC (6 mg/m(2)) on days 7 and 14 and in months 2, 4 and 6, by bolus injection and oral 5-FU (200 mg/day) for 6 months. The postoperative chemotherapy (PC) group received postoperative chemotherapy only. RESULTS: In an intent-to-treat analysis, the 5-year survival rate in the PPC group and the PC group was 77.3% and 75.7% for colon cancer and 67.2% and 69.2% for rectal cancer, respectively. In a per-protocol analysis, the 5-year DFS rate in the PPC group and the PC group was 76.0% and 80.7% for colon cancer and 60.5% and 63.0% for rectal cancer, respectively, indicating no significant differences between the two groups. Adverse reactions were generally mild, confirming the safety of this preoperative chemotherapeutic regimen. CONCLUSION: In the PC group, the 5-year survival rate was nearly identical with that seen in our earlier research using the same regimen, reaffirming the clinical effectiveness of postoperative MMC by protracted intravenous infusion and oral 5-FU. However, our findings did not support additional preoperative chemotherapy for curative resection in patients with colorectal cancer. PMID- 12913084 TI - Phase I study of weekly docetaxel infusion and concurrent radiation therapy for head and neck cancer. AB - OBJECTIVE: This study was designed to determine the maximum tolerated dose (MTD) and dose-limiting toxicity (DLT) of weekly docetaxel in combination with concurrent radiotherapy for treating head and neck cancer. METHODS: Twelve patients with unresectable or postoperative head and neck cancers were enrolled in a dose-escalating phase I study. Eleven of the 12 patients were postoperative patients with intermediate or high pathological risk features. Radiotherapy was delivered as a standard fractionation regimen (2 Gy/day, 5 fractions/week) to a total dose of 60-70 Gy. The starting dose of docetaxel was 10 mg/m(2) (once per week) with a subsequent dose escalation of 5 mg/m(2) in cohorts of three patients. In 2001 and 2002, 12 patients completing three dose levels were included in the study. RESULTS: The MTD of docetaxel was 20 mg/m(2). With the third dose level (20 mg/m(2)), DLT was observed in two of three patients. One experienced grade IV mucositis and another suffered from prolonged grade III mucositis-enforced treatment delay for 13 days. Hematological toxicity was minimal. Mucositis was the DLT of concurrent chemoradiotherapy using weekly docetaxel administration for head and neck cancer. CONCLUSIONS: We identified the recommended phase II dose of docetaxel as 15 mg/m(2) administered weekly with concurrent radiotherapy for head and neck cancers. PMID- 12913085 TI - Phase I and pharmacokinetic study of KRN5500, a spicamycin derivative, for patients with advanced solid tumors. AB - BACKGROUND: KRN5500, a novel spicamycin derivative, shows an inhibitory effect on protein synthesis. This phase I study was aimed at investigating the toxicity, maximum tolerated dose (MTD) and pharmacokinetics of this compound. PATIENTS AND METHODS: Patients with solid tumors not amenable to standard forms of treatment were eligible. KRN5500 was administered as a 2 h intravenous infusion every 4 weeks at doses of 3, 6, 10, 15 and 21 mg/m(2). Pharmacokinetic evaluation was performed at the first cycle. RESULTS: Eighteen patients with advanced solid tumors were enrolled. A total of 26 cycles of KRN5500 were administered. The major toxicities were nausea, vomiting, diarrhea, fatigue and a mild reversible prolongation of prothrombin time. Grade 4 pulmonary toxicity (interstitial pneumonitis) was observed in one patient at a dose level of 15 mg/m(2). Severe fatigue was observed in one patient at a dose level of 21 mg/m(2) and the duration of fatigue tended to increase with the dose of KRN5500. Nausea and vomiting were frequently observed and became prolonged with increasing dose of KRN5500. These toxicity profiles were identified as unacceptable and further dose escalation above 21 mg/m(2) was withheld. The MTD was therefore determined as 21 mg/m(2). The peak plasma concentration and the area under the concentration-time curve of KRN5500 increased proportionally to the dose, suggesting linear pharmacokinetics. No objective antitumor response was observed. CONCLUSION: KRN5500, a structurally novel protein synthesis inhibitor, warrants further investigation to overcome these toxicity profiles and improve its efficacy. PMID- 12913086 TI - Standardization of the body surface area (BSA) formula to calculate the dose of anticancer agents in Japan. AB - BACKGROUND: The importance of deciding the appropriate dose of anticancer agents cannot be overemphasized. Body surface area (BSA) has been used to calculate the dose in anticancer therapy since the 1950s. Japanese oncologists, often use their own Japanese BSA formula instead of western BSA formulae. However, it is not widely known that some discrepancies exist between the BSA products of the Japanese and western styles. On the other hand, recently dose-calculations according to BSA were criticized from the standpoint of pharmacokinetics (PK). Lately, we have had many opportunities for international collaborations, which make it necessary to review these BSA formulae, and the BSA-based dosing method. A unified BSA formula in cancer therapy is needed in Japan. METHODS: We searched and compiled frequently used BSA formulae across the world using the MEDLINE search, oncology text, a web search on cancer clinical trial groups, and personally communicated with medical oncologists from western countries. Using these formulae, we calculated BSA for a typical Japanese individual, and compared their products. We calculated BSA using these formulae for individuals of widely varying physique, from 140 to 185 cm in height, and from 30 to 96 kg in weight, and estimated the amount of discrepancies among them. RESULTS: Among the various BSA formulae used in western countries, the DuBois formula is the standard. In Japan, the Fujimoto formula has been used frequently. The Fujimoto formula was based on a study of 201 Japanese subjects in 1949. For the average Japanese individual, the BSA calculated using the Fujimoto formula was about 3% lower than that which was calculated by western formulae. The BSA calculated for all heights and body weights using the Fujimoto formula, ranged between 0.7 and 4.8% less than those calculated by using the DuBois formula. The other western formulae showed larger discrepancies than the Fujimoto and DuBois formulae. CONCLUSION: BSA-based dosing has failed to standardize the variation in PK for most anticancer agents, and individual dosing techniques are currently being investigated. However, until their clinical utilities are confirmed, it is necessary to depend on the BSA-based calculation for determining the dose of most anticancer agents. The DuBois formula, which is the western standard formula, is validated to a greater extent and its accuracy has been confirmed more than others, including the Fujimoto formula. We recommend the use of the DuBois formula instead of the Fujimoto formula in cancer chemotherapy and propose the standardization of this formula in Japan. PMID- 12913087 TI - Hepatocarcinoma with congenital agenesis of the portal vein. AB - The association of congenital absence of the portal vein and hepatocellular carcinoma has been described, but is rare. It is most frequently discovered fortuitously in children. The absence of intrahepatic portal circulation may predispose to the development of hepatocarcinoma. PMID- 12913088 TI - Fractal properties of perfusion heterogeneity in optimized arterial trees: a model study. AB - Regional blood flows in the heart muscle are remarkably heterogeneous. It is very likely that the most important factor for this heterogeneity is the metabolic need of the tissue rather than flow dispersion by the branching network of the coronary vasculature. To model the contribution of tissue needs to the observed flow heterogeneities we use arterial trees generated on the computer by constrained constructive optimization. This method allows to prescribe terminal flows as independent boundary conditions, rather than obtaining these flows by the dispersive effects of the tree structure. We study two specific cases: equal terminal flows (model 1) and terminal flows set proportional to the volumes of Voronoi polyhedra used as a model for blood supply regions of terminal segments (model 2). Model 1 predicts, depending on the number Nterm of end-points, fractal dimensions D of perfusion heterogeneities in the range 1.20 to 1.40 and positively correlated nearest-neighbor regional flows, in good agreement with experimental data of the normal heart. Although model 2 yields reasonable terminal flows well approximated by a lognormal distribution, it fails to predict D and nearest-neighbor correlation coefficients r1 of regional flows under normal physiologic conditions: model 2 gives D = 1.69 +/- 0.02 and r1 = -0.18 +/- 0.03 (n = 5), independent of Nterm and consistent with experimental data observed under coronary stenosis and under the reduction of coronary perfusion pressure. In conclusion, flow heterogeneity can be modeled by terminal positions compatible with an existing tree structure without resorting to the flow-dispersive effects of a specific branching tree model to assign terminal flows. PMID- 12913089 TI - Gating competence of constitutively open CLC-0 mutants revealed by the interaction with a small organic Inhibitor. AB - Opening of CLC chloride channels is coupled to the translocation of the permeant anion. From the recent structure determination of bacterial CLC proteins in the closed and open configuration, a glutamate residue was hypothesized to form part of the Cl--sensitive gate. The negatively charged side-chain of the glutamate was suggested to occlude the permeation pathway in the closed state, while opening of a single protopore of the double-pore channel would reflect mainly a movement of this side-chain toward the extracellular pore vestibule, with little rearrangement of the rest of the channel. Here we show that mutating this critical residue (Glu166) in the prototype Torpedo CLC-0 to alanine, serine, or lysine leads to constitutively open channels, whereas a mutation to aspartate strongly slowed down opening. Furthermore, we investigated the interaction of the small organic channel blocker p-chlorophenoxy-acetic acid (CPA) with the mutants E166A and E166S. Both mutants were strongly inhibited by CPA at negative voltages with a >200-fold larger affinity than for wild-type CLC-0 (apparent KD at -140 mV approximately 4 micro M). A three-state linear model with an open state, a low affinity and a high-affinity CPA-bound state can quantitatively describe steady state and kinetic properties of the CPA block. The parameters of the model and additional mutagenesis suggest that the high-affinity CPA-bound state is similar to the closed configuration of the protopore gate of wild-type CLC-0. In the E166A mutant the glutamate side chain that occludes the permeation pathway is absent. Thus, if gating consists only in movement of this side-chain the mutant E166A should not be able to assume a closed conformation. It may thus be that fast gating in CLC-0 is more complex than anticipated from the bacterial structures. PMID- 12913090 TI - Conformational changes in the pore of CLC-0. AB - The Torpedo Cl- channel, CLC-0, is inhibited by clofibric acid derivatives from the intracellular side. We used the slow gate-deficient mutant CLC-0C212S to investigate the mechanism of block by the clofibric acid-derivative p chlorophenoxy-acetic acid (CPA). CPA blocks open channels with low affinity (KDO= 45 mM at 0 mV) and shows fast dissociation (koff = 490 s-1 at -140 mV). In contrast, the blocker binds to closed channels with higher affinity and with much slower kinetics. This state-dependent block coupled with the voltage dependence of the gating transitions results in a highly voltage-dependent inhibition of macroscopic currents (KD approximately 1 mM at -140 mV; KD approximately 65 mM at 60 mV). The large difference in CPA affinity of the open and closed state suggests that channel opening involves more than just a local conformational rearrangement. On the other hand, in a recent work (Dutzler, R., E.B. Campbell, and R. MacKinnon. 2003. Science. 300:108-112) it was proposed that the conformational change underlying channel opening is limited to a movement of a single side chain. A prediction of this latter model is that mutations that influence CPA binding to the channel should affect the affinities for an open and closed channel in a similar manner since the general structure of the pore remains largely unchanged. To test this hypothesis we introduced point mutations in four residues (S123, T471, Y512, and K519) that lie close to the intracellular pore mouth or to the putative selectivity filter. Mutation T471S alters CPA binding exclusively to closed channels. Pronounced effects on the open channel block are observed in three other mutants, S123T, Y512A, and K519Q. Together, these results collectively suggest that the structure of the CPA binding site is different in the open and closed state. Finally, replacement of Tyr 512, a residue directly coordinating the central Cl- ion in the crystal structure, with Phe or Ala has very little effect on single channel conductance and selectivity. These observations suggest that channel opening in CLC-0 consists in more than a movement of a side chain and that other parts of the channel and of the selectivity filter are probably involved. PMID- 12913091 TI - State-dependent block of wild-type and inactivation-deficient Na+ channels by flecainide. AB - The antiarrhythmic agent flecainide appears beneficial for painful congenital myotonia and LQT-3/DeltaKPQ syndrome. Both diseases manifest small but persistent late Na+ currents in skeletal or cardiac myocytes. Flecainide may therefore block late Na+ currents for its efficacy. To investigate this possibility, we characterized state-dependent block of flecainide in wild-type and inactivation deficient rNav1.4 muscle Na+ channels (L435W/L437C/A438W) expressed with beta1 subunits in Hek293t cells. The flecainide-resting block at -140 mV was weak for wild-type Na+ channels, with an estimated 50% inhibitory concentration (IC50) of 365 micro M when the cell was not stimulated for 1,000 s. At 100 micro M flecainide, brief monitoring pulses of +30 mV applied at frequencies as low as 1 per 60 s, however, produced an approximately 70% use-dependent block of peak Na+ currents. Recovery from this use-dependent block followed an exponential function, with a time constant over 225 s at -140 mV. Inactivated wild-type Na+ channels interacted with flecainide also slowly at -50 mV, with a time constant of 7.9 s. In contrast, flecainide blocked the open state of inactivation deficient Na+ channels potently as revealed by its rapid time-dependent block of late Na+ currents. The IC50 for flecainide open-channel block at +30 mV was 0.61 micro M, right within the therapeutic plasma concentration range; on-rate and off rate constants were 14.9 micro M-1s-1 and 12.2 s-1, respectively. Upon repolarization to -140 mV, flecainide block of inactivation-deficient Na+ channels recovered, with a time constant of 11.2 s, which was approximately 20 fold faster than that of wild-type counterparts. We conclude that flecainide directly blocks persistent late Na+ currents with a high affinity. The fast inactivation gate, probably via its S6 docking site, may further stabilize the flecainide-receptor complex in wild-type Na+ channels. PMID- 12913092 TI - Molecular motions of the outer ring of charge of the sodium channel: do they couple to slow inactivation? AB - In contrast to fast inactivation, the molecular basis of sodium (Na) channel slow inactivation is poorly understood. It has been suggested that structural rearrangements in the outer pore mediate slow inactivation of Na channels similar to C-type inactivation in potassium (K) channels. We probed the role of the outer ring of charge in inactivation gating by paired cysteine mutagenesis in the rat skeletal muscle Na channel (rNav1.4). The outer charged ring residues were substituted with cysteine, paired with cysteine mutants at other positions in the external pore, and coexpressed with rat brain beta1 in Xenopus oocytes. Dithiolthreitol (DTT) markedly increased the current in E403C+E758C double mutant, indicating the spontaneous formation of a disulfide bond and proximity of the alpha carbons of these residues of no more than 7 A. The redox catalyst Cu(II) (1,10-phenanthroline)3 (Cu(phe)3) reduced the peak current of double mutants (E403C+E758C, E403C+D1241C, E403C+D1532C, and D1241C+D1532C) at a rate proportional to the stimulation frequency. Voltage protocols that favored occupancy of slow inactivation states completely prevented Cu(phe)3 modification of outer charged ring paired mutants E403C+E758C, E403C+D1241C, and E403C+D1532C. In contrast, voltage protocols that favored slow inactivation did not prevent Cu(phe)3 modification of other double mutants such as E403C+W756C, E403C+W1239C, and E403C+W1531C. Our data suggest that slow inactivation of the Na channel is associated with a structural rearrangement of the outer ring of charge. PMID- 12913093 TI - Impaired differentiation of osteoclasts in TREM-2-deficient individuals. AB - TREM-2 is an immunoglobulin-like cell surface receptor associated with DAP12/KARAP that activates monocyte-derived dendritic cells (DCs) in vitro. Recently, it has been shown that genetic defects of human DAP12/KARAP and TREM-2 result in a rare syndrome characterized by bone cysts and presenile dementia called Nasu-Hakola disease. This observation suggests that TREM-2 may function in myeloid cells other than DCs, most probably osteoclasts (OCs) and microglial cells, which are involved in bone modeling and brain function. Consistent with this prediction, here we show that OC differentiation is dramatically arrested in TREM-2-deficient patients, resulting in large aggregates of immature OCs that exhibit impaired bone resorptive activity. These results demonstrate a critical role for TREM-2 in the differentiation of mononuclear myeloid precursors into functional multinucleated OCs. PMID- 12913094 TI - A role of the fast ATP-gated P2X1 cation channel in thrombosis of small arteries in vivo. AB - The P2X1 receptor is a fast ATP-gated cation channel expressed in blood platelets, where its role has been difficult to assess due to its rapid desensitization and the lack of pharmacological tools. In this paper, we have used P2X1-/- and wild-type mouse platelets, treated with apyrase to prevent desensitization, to demonstrate the function of P2X1 in the response to thrombogenic stimuli. In vitro, the collagen-induced aggregation and secretion of P2X1-deficient platelets was decreased, as was adhesion and thrombus growth on a collagen-coated surface, particularly when the wall shear rate was elevated. In vivo, the functional role of P2X1 could be demonstrated using two models of platelet-dependent thrombotic occlusion of small arteries, in which blood flow is characterized by a high shear rate. The mortality of P2X1-/- mice in a model of systemic thromboembolism was reduced and the size of mural thrombi formed after a laser-induced vessel wall injury was decreased as compared with normal mice, whereas the time for complete thrombus removal was shortened. Overall, the P2X1 receptor appears to contribute to the formation of platelet thrombi, particularly in arteries in which shear forces are high. PMID- 12913095 TI - Coupling between B cell receptor and phospholipase C-gamma2 is essential for mature B cell development. AB - Two signaling pathways known to be essential for progression from immature to mature B cells are BAFF receptor (BAFF-R) and the B cell receptor (BCR). Here, we first show that phospholipase C (PLC)-gamma2 is required for a BAFF-R-mediated survival signal. Then, we have examined the question of whether the reduced number of mature B cells in PLC-gamma2-/- mice is caused by a defect in either BCR or BAFF-R signaling. We find that a PLC-gamma2 SH2 mutant, which inhibits coupling between BCR and PLC-gamma2, fails to restore B cell maturation, despite supporting BAFF-dependent survival. Therefore, our data suggest that the BAFF-R mediated survival signal, provided by PLC-gamma2, is not sufficient to promote B cell maturation, and that, in addition, activation of PLC-gamma2 by BCR is required for B cell development. PMID- 12913096 TI - Identification of PVR (CD155) and Nectin-2 (CD112) as cell surface ligands for the human DNAM-1 (CD226) activating molecule. AB - Human natural killer (NK) cells express a series of activating receptors and coreceptors that are involved in recognition and killing of target cells. In this study, in an attempt to identify the cellular ligands for such triggering surface molecules, mice were immunized with NK-susceptible target cells. On the basis of a functional screening, four mAbs were selected that induced a partial down regulation of the NK-mediated cytotoxicity against the immunizing target cells. As revealed by biochemical analysis, three of such mAbs recognized molecules of approximately 70 kD. The other mAb reacted with two distinct molecules of approximately 65 and 60 kD, respectively. Protein purification followed by tryptic digestion and mass spectra analysis, allowed the identification of the 70 kD and the 65/60 kD molecules as PVR (CD155) and Nectin-2 delta/alpha (CD112), respectively. PVR-Fc and Nectin-2-Fc soluble hybrid molecules brightly stained COS-7 cells transfected with the DNAM-1 (CD226) construct, thus providing direct evidence that both PVR and Nectin-2 represent specific ligands for the DNAM-1 triggering receptor. Finally, the surface expression of PVR or Nectin-2 in cell transfectants resulted in DNAM-1-dependent enhancement of NK-mediated lysis of these target cells. This lysis was inhibited or even virtually abrogated upon mAb mediated masking of DNAM-1 (on NK cells) or PVR or Nectin-2 ligands (on cell transfectants). PMID- 12913097 TI - Thallium-201 single-photon emission computed tomography as an early predictor of outcome in recurrent glioma. AB - PURPOSE: With limited response rates and potential toxicity of chemotherapeutic treatment in patients with recurrent glioma, reliable response assessment is essential. Currently, the assessment of treatment response in glioma patients is based on the combination of radiologic and clinical findings. However, response monitoring with computed tomography (CT) or magnetic resonance imaging (MRI) is hampered by several pitfalls and is prone to interobserver variability. The aim of this study was to establish the value of thallium-201 single-photon emission computed tomography (201Tl-SPECT) as a predictor of overall survival and response to chemotherapy in recurrent glioma, and to compare the value of 201Tl-SPECT with that of CT and MRI. PATIENTS AND METHODS: We studied patients who underwent CT or MRI and 201Tl-SPECT before chemotherapy (n = 57), and patients who also had undergone CT or MRI and 201Tl-SPECT after two courses of chemotherapy (n = 44). The value of the radiologic variables (CT-MRI tumor size, 201Tl-SPECT tumor size, and maximal tumor intensity) at baseline and at follow-up in predicting overall survival, and the percentage of patients alive and progression-free at 6 months (APF6) were examined using Cox regression and logistic regression analysis. RESULTS: Both at baseline and at follow-up, 201Tl-SPECT maximal tumor intensity was the strongest predictive variable and was inversely related to overall survival and APF6. In particular, progression of maximal tumor intensity after two courses of chemotherapy was a powerful predictor of poor outcome. CONCLUSION: 201Tl-SPECT is superior to conventional CT-MRI in the early prediction of overall survival and response to chemotherapy in patients with recurrent glioma. PMID- 12913098 TI - Prediction of disease outcome in melanoma patients by molecular analysis of paraffin-embedded sentinel lymph nodes. AB - PURPOSE: A significant number of patients who develop recurrence after a histopathologically negative sentinel lymph node (SLN) biopsy will demonstrate occult metastases on re-evaluation of the SLNs with serial sectioning and immunohistochemistry. Reverse transcriptase polymerase chain reaction (RT-PCR) has been evaluated to improve disease staging and avoid false-negative findings in fresh or frozen-section SLNs. The purpose of this study was to develop a multimarker RT-PCR assay for assessing melanoma patients' archived paraffin embedded (PE) SLNs. PATIENTS AND METHODS: Archived PE histopathologically positive (n = 37) and negative (n = 40) SLNs from patients with primary melanoma were analyzed using a semiquantitative multimarker RT-PCR assay. RESULTS: Marker expression in histopathologically positive and negative SLNs were as follows: 89%, 92%, 35%, and 43% (positive) and 40%, 33%, 5%, and 13% (negative) for tyrosinase, melanoma antigen recognized by T cells-1, tyrosinase-related protein 1, and tyrosinase-related protein-2, respectively. Twenty-five percent of histopathologically negative SLN patients were upstaged using at least two markers. Of these, 80% developed a recurrence. Furthermore, at a median follow-up of 55 months, patients with histopathologically negative SLNs who expressed zero or one marker had a significantly improved disease-free (P <.002) and overall (P <.03) survival versus those expressing two or more markers. CONCLUSION: These findings demonstrate the feasibility of a multimarker RT-PCR assay for evaluating archived PE SLNs. More significantly, identification of molecular risk factors can be detected in histopathologically negative SLNs for distinguishing early stage melanoma patients with a worse prognosis. PMID- 12913099 TI - Undertreatment strongly decreases prognosis of breast cancer in elderly women. AB - PURPOSE: No consensus exists on therapy of elderly cancer patients. Treatments are influenced by unclear standards and are usually less aggressive. This study aims to evaluate determinants and effect of treatment choice on breast cancer prognosis among elderly patients. PATIENTS AND METHODS: We reviewed clinical files of 407 breast cancer patients aged >/= 80 years recorded at the Geneva Cancer Registry between 1989 and 1999. Patient and tumor characteristics, general health status, comorbidity, treatment, and cause of death were considered. We evaluated determinants of treatment by logistic regression and effect of treatment on mortality by Cox model, accounting for prognostic factors. RESULTS: Age was independently linked to the type of treatment. Overall, 12% of women (n = 48) had no treatment, 32% (n = 132) received tamoxifen only, 7% (n = 28) had breast-conserving surgery only, 33% (n = 133) had mastectomy, 14% (n = 57) had breast-conserving surgery plus adjuvant therapy, and 2% (n = 9) received miscellaneous treatments. Five-year specific breast cancer survival was 46%, 51%, 82%, and 90% for women with no treatment, tamoxifen alone, mastectomy, and breast conserving surgery plus adjuvant treatment, respectively. Compared with the nontreated group, the adjusted hazard ratio of breast cancer mortality was 0.4 (95% CI, 0.2 to 0.7) for tamoxifen alone, 0.4 (95% CI, 0.1 to 1.4) for breast conserving surgery alone, 0.2 (95% CI, 0.1 to 0.7) for mastectomy, and 0.1 (95% CI, 0.03 to 0.4) for breast-conserving surgery plus adjuvant treatment. CONCLUSION: Half of elderly patients with breast cancer are undertreated, with strongly decreased specific survival as a consequence. Treatments need to be adapted to the patient's health status, but also should offer the best chance of cure. PMID- 12913101 TI - What constitutes optimal care for older women with breast cancer? PMID- 12913100 TI - Involved-field radiotherapy is equally effective and less toxic compared with extended-field radiotherapy after four cycles of chemotherapy in patients with early-stage unfavorable Hodgkin's lymphoma: results of the HD8 trial of the German Hodgkin's Lymphoma Study Group. AB - PURPOSE: To investigate whether radiotherapy can be reduced without loss of efficacy from extended field (EF) to involved field (IF) after four cycles of chemotherapy. PATIENTS AND METHODS: Between 1993 and 1998, patients with newly diagnosed early-stage unfavorable HD were enrolled onto this multicenter study. Patients were randomly assigned to receive cyclophosphamide, vincristine, procarbazine, and prednisone (COPP) + doxorubicin, bleomycin, vinblastine, and dacarbazine (ABVD) for two cycles followed by radiotherapy of 30 Gy EF + 10 Gy to bulky disease (arm A) or 30 Gy IF + 10 Gy to bulky disease (arm B). RESULTS: Of 1,204 patients randomly assigned to treatment, 1,064 patients were informative and eligible for the arm comparison (532 patients in arm A; 532 patients in arm B). The median observation time was 54 months. Five years after random assignment, the overall survival (OSran) for all eligible patients was 91% and freedom from treatment failure (FFTFran) was 83%. Survival rates at 5 years after start of radiotherapy revealed no differences for arms A and B, respectively, in terms of FFTF (85.8% and 84.2%) and OS at 5 years (90.8% and 92.4%). There also were no differences between arms A and B, respectively, in terms of complete remission (98.5% and 97.2%), progressive disease (0.8% and 1.9%), relapse (6.4% and 7.7%), death (8.1% and 6.4%), and secondary neoplasia (4.5% and 2.8%). In contrast, acute side effects including leukopenia, thrombocytopenia, nausea, gastrointestinal toxicity, and pharyngeal toxicity were more frequent in the EF arm. CONCLUSION: Radiotherapy volume size reduction from EF to IF after COPP + ABVD chemotherapy for two cycles produces similar results and less toxicity in patients with early-stage unfavorable HD. PMID- 12913102 TI - Glioma therapies: how to tell which work? PMID- 12913103 TI - The use of the mini-mental state examination to assess cognitive functioning in cancer trials: no ifs, ands, buts, or sensitivity. PMID- 12913104 TI - The ingredients for prostate cancer nomograms: the addition of biomarkers sets the table for future recipes. PMID- 12913105 TI - Advances in molecular staging of melanoma patients: multimarker analysis of archival lymph node tissue. PMID- 12913106 TI - The addition of interleukin-6 soluble receptor and transforming growth factor beta1 improves a preoperative nomogram for predicting biochemical progression in patients with clinically localized prostate cancer. AB - PURPOSE: Several preoperative prostate cancer nomograms have been developed that predict risk of progression using pretreatment prostate-specific antigen (PSA) level, clinical stage, and biopsy Gleason grade. We describe the development and performance of a new nomogram. The nomogram adds new markers to the standard clinical predictors that reflect the biologic behavior of prostate cancer: pretreatment plasma levels of interleukin-6 soluble receptor (IL6SR) and transforming growth factor beta1 (TGF-beta1). PATIENTS AND METHODS: Between November 7, 1994 and December 22, 1997, 714 patients with stage cT1c to cT3a prostate cancer and no prior therapy were treated with radical prostatectomy at the Methodist Hospital, Houston TX. Plasma levels of IL6SR and TGF-beta1 were measured in banked preoperative plasma. With these data, a nomogram was developed to predict the probability of PSA progression within 5 years of surgery. The nomogram was validated with bootstrapping to assess its discrimination and calibration performance. RESULTS: In the multivariable Cox model, PSA (P =.004), IL6SR (P <.001), TGF-beta1 (P <.001), primary Gleason grade (P <.002), and secondary Gleason grade (P =.029) were associated with PSA progression, whereas clinical stage (P =.696) was not. The nomogram seemed to be well calibrated and had a bootstrap-corrected area under the receiver operating characteristic curve (ie, concordance index) of 0.83. For comparison, a nomogram that omitted IL6SR and TGF-beta1 achieved a concordance index of only 0.75. CONCLUSION: We found that pretreatment plasma levels of IL6SR and TGF-beta1 improved the ability to predict biochemical progression by a prognostically substantial margin. A nomogram including the pretreatment levels of these molecular markers, along with standard clinical markers, has been developed and internally validated. PMID- 12913107 TI - Unique targeting of cytosolic phospholipase A2 to plasma membranes mediated by the NADPH oxidase in phagocytes. AB - Cytosolic phospholipase A2 (cPLA2)-generated arachidonic acid (AA) has been shown to be an essential requirement for the activation of NADPH oxidase, in addition to its being the major enzyme involved in the formation of eicosanoid at the nuclear membranes. The mechanism by which cPLA2 regulates NADPH oxidase activity is not known, particularly since the NADPH oxidase complex is localized in the plasma membranes of stimulated cells. The present study is the first to demonstrate that upon stimulation cPLA2 is transiently recruited to the plasma membranes by a functional NADPH oxidase in neutrophils and in granulocyte-like PLB-985 cells. Coimmunoprecipitation experiments and double labeling immunofluorescence analysis demonstrated the unique colocalization of cPLA2 and the NADPH oxidase in plasma membranes of stimulated cells, in correlation with the kinetic burst of superoxide production. A specific affinity in vitro binding was detected between GST-p47phox or GST-p67phox and cPLA2 in lysates of stimulated cells. The association between these two enzymes provides the molecular basis for AA released by cPLA2 to activate the assembled NADPH oxidase. The ability of cPLA2 to regulate two different functions in the same cells (superoxide generation and eicosanoid production) is achieved by a novel dual subcellular localization of cPLA2 to different targets. PMID- 12913108 TI - The GAP activity of Msb3p and Msb4p for the Rab GTPase Sec4p is required for efficient exocytosis and actin organization. AB - Polarized growth in Saccharomyces cerevisiae is thought to occur by the transport of post-Golgi vesicles along actin cables to the daughter cell, and the subsequent fusion of the vesicles with the plasma membrane. Previously, we have shown that Msb3p and Msb4p genetically interact with Cdc42p and display a GTPase activating protein (GAP) activity toward a number of Rab GTPases in vitro. We show here that Msb3p and Msb4p regulate exocytosis by functioning as GAPs for Sec4p in vivo. Cells lacking the GAP activity of Msb3p and Msb4p displayed secretory defects, including the accumulation of vesicles of 80-100 nm in diameter. Interestingly, the GAP activity of Msb3p and Msb4p was also required for efficient polarization of the actin patches and for the suppression of the actin-organization defects in cdc42 mutants. Using a strain defective in polarized secretion and actin-patch organization, we showed that a change in actin-patch organization could be a consequence of the fusion of mistargeted vesicles with the plasma membrane. PMID- 12913109 TI - Phosphatidylinositol phosphate 5-kinase Ibeta recruits AP-2 to the plasma membrane and regulates rates of constitutive endocytosis. AB - Overexpression of phosphatidylinositol phosphate 5-kinase (PIP5KI) isoforms alpha, beta, or gamma in CV-1 cells increased phosphatidylinositol 4,5 bisphosphate (PIP2) levels by 35, 180, and 0%, respectively. Endocytosis of transferrin receptors, association of AP-2 proteins with membranes, and the number of clathrin-coated pits at the plasma membrane increased when PIP2 increased. When expression of PIP5KIbeta was inhibited with small interference RNA in HeLa cells, expression of PIP5KIalpha was also reduced slightly, but PIP5KIgamma expression was increased. PIP2 levels and internalization of transferrin receptors dropped 50% in these cells; thus, PIP5KIgamma could not compensate for loss of PIP5KIbeta. When expression of PIP5KIalpha was reduced, expression of both PIP5KIbeta and PIP5KIgamma increased and PIP2 levels did not change. A similar increase of PIP5KIalpha and PIP5KIbeta occurred when PIP5KIgamma was inhibited. These results indicate that constitutive endocytosis in CV-1 and HeLa cells requires (and may be regulated by) PIP2 produced primarily by PIP5KIbeta. PMID- 12913111 TI - The tyrosine phosphatase CD148 is excluded from the immunologic synapse and down regulates prolonged T cell signaling. AB - CD148 is a receptor-like protein tyrosine phosphatase up-regulated on T cells after T cell receptor (TCR) stimulation. To examine the physiologic role of CD148 in TCR signaling, we used an inducible CD148-expressing Jurkat T cell clone. Expression of CD148 inhibits NFAT (nuclear factor of activated T cells) activation induced by soluble anti-TCR antibody, but not by antigen-presenting cells (APCs) loaded with staphylococcal enterotoxin superantigen (SAg) or immobilized anti-TCR antibody. Immunofluorescence microscopy revealed that the extracellular domain of CD148 mediates its exclusion from the immunologic synapse, sequestering it from potential substrates. Targeting of the CD148 phosphatase domain to the immunologic synapse potently inhibited NFAT activation by all means of triggering through the TCR. These data lead us to propose a model where CD148 function is regulated in part by exclusion from substrates in the immunologic synapse. Upon T cell-APC disengagement, CD148 can then access and dephosphorylate substrates to down-regulate prolongation of signaling. PMID- 12913110 TI - FOXO transcription factors directly activate bim gene expression and promote apoptosis in sympathetic neurons. AB - Developing sympathetic neurons die by apoptosis when deprived of NGF. BIM, a BH3 only member of the BCL-2 family, is induced after NGF withdrawal in these cells and contributes to NGF withdrawal-induced death. Here, we have investigated the involvement of the Forkhead box, class O (FOXO) subfamily of Forkhead transcription factors in the regulation of BIM expression by NGF. We find that overexpression of FOXO transcription factors induces BIM expression and promotes death of sympathetic neurons in a BIM-dependent manner. In addition, we find that FKHRL1 (FOXO3a) directly activates the bim promoter via two conserved FOXO binding sites and that mutation of these sites abolishes bim promoter activation after NGF withdrawal. Finally, we show that FOXO activity contributes to the NGF deprivation-induced death of sympathetic neurons. PMID- 12913112 TI - Dual recognition of the ribosome and the signal recognition particle by the SRP receptor during protein targeting to the endoplasmic reticulum. AB - We have analyzed the interactions between the signal recognition particle (SRP), the SRP receptor (SR), and the ribosome using GTPase assays, biosensor experiments, and ribosome binding assays. Possible mechanisms that could contribute to an enhanced affinity between the SR and the SRP-ribosome nascent chain complex to promote protein translocation under physiological ionic strength conditions have been explored. Ribosomes or 60S large ribosomal subunits activate the GTPase cycle of SRP54 and SRalpha by providing a platform for assembly of the SRP-SR complex. Biosensor experiments revealed high-affinity, saturable binding of ribosomes or large ribosomal subunits to the SR. Remarkably, the SR has a 100 fold higher affinity for the ribosome than for SRP. Proteoliposomes that contain the SR bind nontranslating ribosomes with an affinity comparable to that shown by the Sec61 complex. An NH2-terminal 319-residue segment of SRalpha is necessary and sufficient for binding of SR to the ribosome. We propose that the ribosome-SR interaction accelerates targeting of the ribosome nascent chain complex to the RER, while the SRP-SR interaction is crucial for maintaining the fidelity of the targeting reaction. PMID- 12913113 TI - Spatial restriction of alpha4 integrin phosphorylation regulates lamellipodial stability and alpha4beta1-dependent cell migration. AB - Integrins coordinate spatial signaling events essential for cell polarity and directed migration. Such signals from alpha4 integrins regulate cell migration in development and in leukocyte trafficking. Here, we report that efficient alpha4 mediated migration requires spatial control of alpha4 phosphorylation by protein kinase A, and hence localized inhibition of binding of the signaling adaptor, paxillin, to the integrin. In migrating cells, phosphorylated alpha4 accumulated along the leading edge. Blocking alpha4 phosphorylation by mutagenesis or by inhibition of protein kinase A drastically reduced alpha4-dependent migration and lamellipodial stability. alpha4 phosphorylation blocks paxillin binding in vitro; we now find that paxillin and phospho-alpha4 were in distinct clusters at the leading edge of migrating cells, whereas unphosphorylated alpha4 and paxillin colocalized along the lateral edges of those cells. Furthermore, enforced paxillin association with alpha4 inhibits migration and reduced lamellipodial stability. These results show that topographically specific integrin phosphorylation can control cell migration and polarization by spatial segregation of adaptor protein binding. PMID- 12913114 TI - Gene expression during ER stress-induced apoptosis in neurons: induction of the BH3-only protein Bbc3/PUMA and activation of the mitochondrial apoptosis pathway. AB - Endoplasmic reticulum (ER) stress has been implicated in the pathogenesis of ischemic and neurodegenerative disorders. Treatment of human SH-SY5Y neuroblastoma cells with tunicamycin, an inhibitor of protein glycosylation, rapidly induced the expression of target genes of the unfolded protein response. However, prolonged treatment also triggered a delayed, caspase-dependent cell death. Microarray analysis of gene expression changes during tunicamycin-induced apoptosis revealed that the Bcl-2 homology domain 3-only family member, Bcl-2 binding component 3/p53 upregulated modulator of apoptosis (Bbc3/PUMA), was the most strongly induced pro-apoptotic gene. Expression of Bbc3/PUMA correlated with a Bcl-xL-sensitive release of cytochrome c and the activation of caspase-9 and 3. Increased expression of Bbc3/PUMA was also observed in p53-deficient human cells, in response to the ER stressor thapsigargin, and in rat hippocampal neurons after transient forebrain ischemia. Overexpression of Bbc3/PUMA was sufficient to trigger apoptosis in SH-SY5Y neuroblastoma cells, and human cells deficient in Bbc3/PUMA showed dramatically reduced apoptosis in response to ER stress. Our data suggest that the transcriptional induction of Bbc3/PUMA may be sufficient and necessary for ER stress-induced apoptosis. PMID- 12913115 TI - tRNA synthetase paralogs: evolutionary links in the transition from tRNA dependent amino acid biosynthesis to de novo biosynthesis. PMID- 12913116 TI - Neurodegenerative tauopathy in the worm. PMID- 12913117 TI - Sex peptide and the sperm effect in Drosophila melanogaster. PMID- 12913118 TI - A nuclear target for interleukin-1alpha: interaction with the growth suppressor necdin modulates proliferation and collagen expression. AB - There is growing evidence for the intracellular role of cytokines and growth factors, but the pathways by which these activities occur remain largely obscure. Previous work from our laboratory identified the constitutive, aberrant expression of the 31-kDa IL-1 alpha precursor (pre-IL-1 alpha) in the nuclei of fibroblasts from the lesional skin of patients with systemic sclerosis (SSc). We established that pre-IL-1 alpha expression was associated with increased fibroblast proliferation and collagen production. Further investigation has led to the identification of a mechanism by which nuclear expression of pre-IL-1 alpha affects fibroblast growth and matrix production. By using a yeast two hybrid method, we found that pre-IL-1 alpha binds necdin, a nuclear protein with growth suppressor activity. We mapped the region of pre-IL-1 alpha responsible for necdin binding and found it to be localized near the N terminus, a region that is present on pre-IL-1 alpha, but not the mature 17-kDa cytokine. Expression studies demonstrated that pre-IL-1 alpha associates with necdin in the nuclei of mammalian cell lines and regulates cell growth and collagen expression. Our results provide the first evidence, to our knowledge, of a nuclear target for pre IL-1 alpha. Based on these findings, we propose that the constitutively up regulated expression of pre-IL-1 alpha in the nuclei of SSc fibroblasts up regulates proliferation and matrix production of SSc fibroblasts through binding necdin, and by counteracting its effects on cell growth and collagen production. PMID- 12913120 TI - How the mouse got its stripes. PMID- 12913119 TI - Stem cells of the skin epithelium. AB - Tissue stem cells form the cellular base for organ homeostasis and repair. Stem cells have the unusual ability to renew themselves over the lifetime of the organ while producing daughter cells that differentiate into one or multiple lineages. Difficult to identify and characterize in any tissue, these cells are nonetheless hotly pursued because they hold the potential promise of therapeutic reprogramming to grow human tissue in vitro, for the treatment of human disease. The mammalian skin epithelium exhibits remarkable turnover, punctuated by periods of even more rapid production after injury due to burn or wounding. The stem cells responsible for supplying this tissue with cellular substrate are not yet easily distinguishable from neighboring cells. However, in recent years a significant body of work has begun to characterize the skin epithelial stem cells, both in tissue culture and in mouse and human skin. Some epithelial cells cultured from skin exhibit prodigious proliferative potential; in fact, for >20 years now, cultured human skin has been used as a source of new skin to engraft onto damaged areas of burn patients, representing one of the first therapeutic uses of stem cells. Cell fate choices, including both self-renewal and differentiation, are crucial biological features of stem cells that are still poorly understood. Skin epithelial stem cells represent a ripe target for research into the fundamental mechanisms underlying these important processes. PMID- 12913121 TI - Human speech and birdsong: communication and the social brain. PMID- 12913123 TI - Genetic footprints of demographic expansion in North America, but not Amazonia, during the Late Quaternary. AB - The biotic consequences of climate change have attracted considerable attention. In particular, the "refugial debate" centers on the possible retraction of habitats to limited areas that may have served as refuges for many associated species, especially during glaciations of the Quaternary. One prediction of such scenarios is that populations must have experienced substantial growth accompanying climatic amelioration and the occupation of newly expanded habitats. We used coalescence theory to examine the genetic evidence, or lack thereof, for late Pleistocene refugia of boreal North American and tropical Amazonian mammals. We found substantial and concordant evidence of demographic expansion in North American mammals, particularly at higher latitudes. In contrast, small mammals from western Amazonia appear to have experienced limited or no demographic expansion after the Late Pleistocene. Thus, demographic responses to climate change can be tracked genetically and appear to vary substantially across the latitudinal gradient of biotic diversity. PMID- 12913122 TI - Protein grafting of an HIV-1-inhibiting epitope. AB - Protein grafting, the transfer of a binding epitope of one ligand onto the surface of another protein, is a potentially powerful technique for presenting peptides in preformed and active three-dimensional conformations. Its utility, however, has been limited by low biological activity of the designed ligands and low tolerance of the protein scaffolds to surface substitutions. Here, we graft the complete binding epitope (19 nonconsecutive amino acids with a solvent accessible surface area of >2,000 A2) of an HIV-1 C-peptide, which is derived from the C-terminal region of HIV-1 gp41 and potently inhibits HIV-1 entry into cells, onto the surface of a GCN4 leucine zipper. The designed peptide, named C34coil, displays a potent antiviral activity approaching that of the native ligand. Moreover, whereas the linear C-peptide is unstructured and sensitive to degradation by proteases, C34coil is well structured, conformationally stable, and exhibits increased resistance to proteolytic degradation compared with the linear peptide. In addition to being a structured antiviral inhibitor, C34coil may also serve as the basis for the development of an alternative class of immunogens. This study demonstrates that "one-shot" protein grafting, without subsequent rounds of optimization, can be used to create ligands with structural conformations and improved biomedical properties. PMID- 12913124 TI - Insights into the mode of inhibition of human mitochondrial monoamine oxidase B from high-resolution crystal structures. AB - Monoamine oxidase B (MAO-B) is an outer mitochondrial membrane-bound enzyme that catalyzes the oxidative deamination of arylalkylamine neurotransmitters and has been a target for a number of clinically used drug inhibitors. The 1.7-A structure of the reversible isatin-MAO-B complex has been determined; it forms a basis for the interpretation of the enzyme's structure when bound to either reversible or irreversible inhibitors. 1,4-Diphenyl-2-butene is found to be a reversible MAO-B inhibitor, which occupies both the entrance and substrate cavity space in the enzyme. Comparison of these two structures identifies Ile-199 as a "gate" between the two cavities. Rotation of the side chain allows for either separation or fusion of the two cavities. Inhibition of the enzyme with N-(2 aminoethyl)-p-chlorobenzamide results in the formation of a covalent N(5) flavin adduct with the phenyl ring of the inhibitor occupying a position in the catalytic site overlapping that of isatin. Inhibition of MAO-B with the clinically used trans-2-phenylcyclopropylamine results in the formation of a covalent C(4a) flavin adduct with an opened cyclopropyl ring and the phenyl ring in a parallel orientation to the flavin. The peptide bond between the flavin substituted Cys-397 and Tyr-398 is in a cis conformation, which allows the proper orientation of the phenolic ring of Tyr-398 in the active site. The flavin ring exists in a twisted nonplanar conformation, which is observed in the oxidized form as well as in both the N(5) and the C(4a) adducts. An immobile water molecule is H-bonded to Lys-296 and to the N(5) of the flavin as observed in other flavin-dependent amine oxidases. The active site cavities are highly apolar; however, hydrophilic areas exist near the flavin and direct the amine moiety of the substrate for binding and catalysis. Small conformational changes are observed on comparison of the different inhibitor-enzyme complexes. Future MAO-B drug design will need to consider "induced fit" contributions as an element in ligand-enzyme interactions. PMID- 12913126 TI - Averaging and renormalization for the Korteveg-deVries-Burgers equation. AB - We consider traveling wave solutions of the Korteveg-deVries-Burgers equation and set up an analogy between the spatial averaging of these traveling waves and real space renormalization for Hamiltonian systems. The result is an effective equation that reproduces means of the unaveraged, highly oscillatory, solution. The averaging enhances the apparent diffusion, creating an "eddy" (or renormalized) diffusion coefficient; the relation between the eddy diffusion coefficient and the original diffusion coefficient is found numerically to be one of incomplete similarity, setting up an instance of Barenblatt's renormalization group. The results suggest a relation between self-similar solutions of differential equations on one hand and renormalization groups and optimal prediction algorithms on the other. An analogy with hydrodynamics is pointed out. PMID- 12913127 TI - Coexistence of periodic modulation of quasiparticle states and superconductivity in Bi2Sr2CaCu2O8+delta. AB - In this article we show, using scanning tunneling spectroscopy, the existence of static striped density of electronic states in nearly optimally doped Bi2Sr2CaCu2O8+delta in zero field. This modulation is aligned with the Cu-O bonds, with a periodicity of four lattice constants, and exhibits features characteristic of a two-dimensional system of line objects. We further show that the density of states modulation manifests itself as a shift of states from above to below the superconducting gap. The fact that a single energy scale (i.e., the gap) appears for both superconductivity and stripes suggests that these two effects have the same origin. PMID- 12913125 TI - A modular, bifunctional RNA that integrates itself into a target RNA. AB - Nature often combines independent functional domains to achieve complex function, but this approach has not been extensively explored with artificial enzymes. Here, a group I ribozyme, which can act as an endoribonuclease, was partnered with the R3C ribozyme, which catalyzes the ligation of RNA molecules. The conjoined ribozymes have the potential to perform successive RNA cleavage and joining reactions, resulting in their mutual integration into a target RNA substrate. When simply joined together, however, the ribozymes were unable to achieve this outcome because of inefficient transfer of the substrate between the two catalytic subunits. In vitro evolution was used to optimize the behavior of the conjoined ribozymes, resulting in bifunctional molecules with substantially improved integration activity. The ligase subunit of these molecules was unchanged, whereas the group I subunit acquired several mutations, mostly in peripheral regions. The generation and study of this bifunctional assembly helps shed light on the evolution of modular enzymes and the obstacles that must be overcome in bringing together independent functional domains. These molecules also may be useful as tools for the insertional mutagenesis of target mRNAs. PMID- 12913128 TI - Malaria in Britain: past, present, and future. AB - There has been much recent speculation that global warming may allow the reestablishment of malaria transmission in previously endemic areas such as Europe and the United States. In this report we analyze temporal trends in malaria in Britain between 1840 and 1910, to assess the potential for reemergence of the disease. Our results demonstrate that at least 20% of the drop-off in malaria was due to increasing cattle population and decreasing acreage of marsh wetlands. Although both rainfall and average temperature were associated with year-to-year variability in death rates, there was no evidence for any association with the long-term malaria trend. Model simulations for future scenarios in Britain suggest that the change in temperature projected to occur by 2050 is likely to cause a proportional increase in local malaria transmission of 8-14%. The current risk is negligible, as >52,000 imported cases since 1953 have not led to any secondary cases. The projected increase in proportional risk is clearly insufficient to lead to the reestablishment of endemicity. PMID- 12913129 TI - Inositol hexakisphosphate mobilizes an endomembrane store of calcium in guard cells. AB - myo-Inositol hexakisphosphate (InsP6) is the most abundant inositol phosphate in cells, yet it remains the most enigmatic of this class of signaling molecule. InsP6 plays a role in the processes by which the drought stress hormone abscisic acid (ABA) induces stomatal closure, conserving water and ensuring plant survival. Previous work has shown that InsP6 levels in guard cells are elevated in response to ABA, and InsP6 inactivates the plasma membrane inward K+ conductance (IK,in) in a cytosolic calcium-dependent manner. The use of laser scanning confocal microscopy in dye-loaded patch-clamped guard cell protoplasts shows that release of InsP6 from a caged precursor mobilizes calcium. Measurement of calcium (barium) currents ICa in patch-clamped protoplasts in whole cell mode shows that InsP6 has no effect on the calcium-permeable channels in the plasma membrane activated by ABA. The InsP6-mediated inhibition of IK,in can also be observed in the absence of external calcium. Thus the InsP6-induced increase in cytoplasmic calcium does not result from calcium influx but must arise from InsP6 triggered release of calcium from endomembrane stores. Measurements of vacuolar currents in patch-clamped isolated vacuoles in whole-vacuole mode showed that InsP6 activates both the fast and slow conductances of the guard cell vacuole. These data define InsP6 as an endomembrane-acting calcium-release signal in guard cells; the vacuole may contribute to InsP6-triggered Ca2+ release, but other endomembranes may also be involved. PMID- 12913130 TI - Stress under the sun: spotlight on ultraviolet-B responses. PMID- 12913132 TI - Gene expression profiling in response to ultraviolet radiation in maize genotypes with varying flavonoid content. AB - Microarray hybridization was used to assess acclimation responses to four UV regimes by near isogenic maize (Zea mays) lines varying in flavonoid content. We found that 355 of the 2,500 cDNAs tested were regulated by UV radiation in at least one genotype. Among these, 232 transcripts are assigned putative functions, whereas 123 encode unknown proteins. UV-B increased expression of stress response and ribosomal protein genes, whereas photosynthesis-associated genes were down regulated; lines lacking UV-absorbing pigments had more dramatic responses than did lines with these pigments, confirming the shielding role of these compounds. Sunlight filtered to remove UV-B or UV-B plus UV-A resulted in significant expression changes in many genes not previously associated with UV responses. Some pathways regulated by UV radiation are shared with defense, salt, and oxidative stresses; however, UV-B radiation can activate additional pathways not shared with other stresses. PMID- 12913133 TI - Convergent responses to stress. Solar ultraviolet-B radiation and Manduca sexta herbivory elicit overlapping transcriptional responses in field-grown plants of Nicotiana longiflora. AB - The effects of solar ultraviolet (UV)-B (280-315 nm) on plants have been studied intensively over the last 2 decades in connection with research on the biological impacts of stratospheric ozone depletion. However, the molecular mechanisms that mediate plant responses to solar (ambient) UV-B and their interactions with response mechanisms activated by other stressors remain for the most part unclear. Using a microarray enriched in wound- and insect-responsive sequences, we examined expression responses of 241 genes to ambient UV-B in field-grown plants of Nicotiana longiflora Cav. Approximately 20% of the sequences represented on the array showed differential expression in response to solar UV B. The expression responses to UV-B had parallels with those elicited by simulated Manduca sexta herbivory. The most obvious similarities were: (a) down regulation of several photosynthesis-related genes, and (b) up-regulation of genes involved in fatty acid metabolism and oxylipin biosynthesis such as HPL (hydroperoxide lyase), alpha-DIOX (alpha-dioxygenase), LOX (13-lipoxygenase), and AOS (allene oxide synthase). Genes encoding a WRKY transcription factor, a ferredoxin-dependent glutamate-synthase, and several other insect-responsive genes of unknown function were also similarly regulated by UV-B and insect herbivory treatments. Our results suggest that UV-B and caterpillar herbivory activate common regulatory elements and provide a platform for understanding the mechanisms of UV-B impacts on insect herbivory that have been documented in recent field studies. PMID- 12913131 TI - Convergence of signaling pathways induced by systemin, oligosaccharide elicitors, and ultraviolet-B radiation at the level of mitogen-activated protein kinases in Lycopersicon peruvianum suspension-cultured cells. AB - We tested whether signaling pathways induced by systemin, oligosaccharide elicitors (OEs), and ultraviolet (UV)-B radiation share common components in Lycopersicon peruvianum suspension-cultured cells. These stress signals all induce mitogen-activated protein kinase (MAPK) activity. In desensitization assays, we found that pretreatment with systemin and OEs transiently reduced the MAPK response to a subsequent treatment with the same or a different elicitor. In contrast, MAPK activity in response to UV-B increased after pretreatment with systemin and OEs. These experiments demonstrate the presence of signaling components that are shared by systemin, OEs, and UV-B. Based on desensitization assays, it is not clear if the same or different MAPKs are activated by different stress signals. To identify specific stress-responsive MAPKs, we cloned three MAPKs from a tomato (Lycopersicon esculentum) leaf cDNA library, generated member specific antibodies, and performed immunocomplex kinase assays with extracts from elicited L. peruvianum cells. Two highly homologous MAPKs, LeMPK1 and LeMPK2, were activated in response to systemin, four different OEs, and UV-B radiation. An additional MAPK, LeMPK3, was only activated by UV-B radiation. The common activation of LeMPK1 and LeMPK2 by many stress signals is consistent with the desensitization assays and may account for substantial overlaps among stress responses. On the other hand, MAPK activation kinetics in response to elicitors and UV-B differed substantially, and UV-B activated a different set of LeMPKs than the elicitors. These differences may account for UV-B-specific responses. PMID- 12913134 TI - Going to "great lengths" to prevent the escape of genes that produce specialty chemicals. PMID- 12913135 TI - Ensuring access to the outcomes of community resource projects. PMID- 12913136 TI - If homogalacturonan were a side chain of rhamnogalacturonan I. Implications for cell wall architecture. PMID- 12913137 TI - The role of phylogenetics in comparative genetics. PMID- 12913138 TI - Interallelic complementation at the ubiquitous urease coding locus of soybean. AB - Soybean (Glycine max [L.] Merrill) mutant aj6 carries a single recessive lesion, aj6, that eliminates ubiquitous urease activity in leaves and callus while retaining normal embryo-specific urease activity. Consistently, aj6/aj6 plants accumulated urea in leaves. In crosses of aj6/aj6 by urease mutants at the Eu1, Eu2, and Eu3 loci, F(1) individuals exhibited wild-type leaf urease activity, and the F(2) segregated urease-negative individuals, demonstrating that aj6 is not an allele at these loci. F(2) of aj6/aj6 crossed with a null mutant lacking the Eu1 encoded embryo-specific urease showed that ubiquitous urease was also inactive in seeds of aj6/aj6. The cross of aj6/aj6 to eu4/eu4, a mutant previously assigned to the ubiquitous urease structural gene (R.S. Torisky, J.D. Griffin, R.L. Yenofsky, J.C. Polacco [1994] Mol Gen Genet 242: 404-414), yielded an F(1) having 22% +/- 11% of wild-type leaf urease activity. Coding sequences for ubiquitous urease were cloned by reverse transcriptase-polymerase chain reaction from wild type, aj6/aj6, and eu4/eu4 leaf RNA. The ubiquitous urease had an 837-amino acid open reading frame (ORF), 87% identical to the embryo-specific urease. The aj6/aj6 ORF showed an R201C change that cosegregated with the lack of leaf urease activity in a cross against a urease-positive line, whereas the eu4/eu4 ORF showed a G468E change. Heteroallelic interaction in F(2) progeny of aj6/aj6 x eu4/eu4 resulted in partially restored leaf urease activity. These results confirm that aj6/aj6 and eu4/eu4 are mutants affected in the ubiquitous urease structural gene. They also indicate that radical amino acid changes in distinct domains can be partially compensated in the urease heterotrimer. PMID- 12913139 TI - RNA expression profiles and data mining of sugarcane response to low temperature. AB - Tropical and subtropical plants are generally sensitive to cold and can show appreciable variation in their response to cold stress when exposed to low positive temperatures. Using nylon filter arrays, we analyzed the expression profile of 1,536 expressed sequence tags (ESTs) of sugarcane (Saccharum sp. cv SP80-3280) exposed to cold for 3 to 48 h. Thirty-four cold-inducible ESTs were identified, of which 20 were novel cold-responsive genes that had not previously been reported as being cold inducible, including cellulose synthase, ABI3 interacting protein 2, a negative transcription regulator, phosphate transporter, and others, as well as several unknown genes. In addition, 25 ESTs were identified as being down-regulated during cold exposure. Using a database of cold regulated proteins reported for other plants, we searched for homologs in the sugarcane EST project database (SUCEST), which contains 263,000 ESTs. Thirty three homologous putative cold-regulated proteins were identified in the SUCEST database. On the basis of the expression profiles of the cold-inducible genes and the data-mining results, we propose a molecular model for the sugarcane response to low temperature. PMID- 12913141 TI - Subcellular targeting of nine calcium-dependent protein kinase isoforms from Arabidopsis. AB - Calcium-dependent protein kinases (CDPKs) are specific to plants and some protists. Their activation by calcium makes them important switches for the transduction of intracellular calcium signals. Here, we identify the subcellular targeting potentials for nine CDPK isoforms from Arabidopsis, as determined by expression of green fluorescent protein (GFP) fusions in transgenic plants. Subcellular locations were determined by fluorescence microscopy in cells near the root tip. Isoforms AtCPK3-GFP and AtCPK4-GFP showed a nuclear and cytosolic distribution similar to that of free GFP. Membrane fractionation experiments confirmed that these isoforms were primarily soluble. A membrane association was observed for AtCPKs 1, 7, 8, 9, 16, 21, and 28, based on imaging and membrane fractionation experiments. This correlates with the presence of potential N terminal acylation sites, consistent with acylation as an important factor in membrane association. All but one of the membrane-associated isoforms targeted exclusively to the plasma membrane. The exception was AtCPK1-GFP, which targeted to peroxisomes, as determined by covisualization with a peroxisome marker. Peroxisome targeting of AtCPK1-GFP was disrupted by a deletion of two potential N terminal acylation sites. The observation of a peroxisome-located CDPK suggests a mechanism for calcium regulation of peroxisomal functions involved in oxidative stress and lipid metabolism. PMID- 12913140 TI - Microarray analysis of the genome-wide response to iron deficiency and iron reconstitution in the cyanobacterium Synechocystis sp. PCC 6803. AB - A full-genome microarray of the (oxy)photosynthetic cyanobacterium Synechocystis sp. PCC 6803 was used to identify genes that were transcriptionally regulated by growth in iron (Fe)-deficient versus Fe-sufficient media. Transcript accumulation for 3,165 genes in the genome was analyzed using an analysis of variance model that accounted for slide and replicate (random) effects and dye (a fixed) effect in testing for differences in the four time periods. We determined that 85 genes showed statistically significant changes in the level of transcription (P 1.25 x). The genes identified included those known previously to be Fe regulated, such as isiA that encodes a novel chlorophyll binding protein responsible for the pigment characteristics of low-Fe (LoFe) cells. ATP synthetase and phycobilisome genes were down-regulated in LoFe, and there were interesting changes in the transcription of genes involved in chlorophyll biosynthesis, in photosystem I and II assembly, and in energy metabolism. Hierarchical clustering demonstrated that photosynthesis genes, as a class, were repressed in LoFe and induced upon the re-addition of Fe. Specific regulatory genes were transcriptionally active in LoFe, including two genes that show homology to plant phytochromes (cph1 and cph2). These observations established the existence of a complex network of regulatory interactions and coordination in response to Fe availability. PMID- 12913142 TI - TaVRT-1, a putative transcription factor associated with vegetative to reproductive transition in cereals. AB - The molecular genetics of vernalization, defined as the promotion of flowering by cold treatment, is still poorly understood in cereals. To better understand this mechanism, we cloned and characterized a gene that we named TaVRT-1 (wheat [Triticum aestivum] vegetative to reproductive transition-1). Molecular and sequence analyses indicated that this gene encodes a protein homologous to the MADS-box family of transcription factors that comprises certain flowering control proteins in Arabidopsis. Mapping studies have localized this gene to the Vrn-1 regions on the long arms of homeologous group 5 chromosomes, regions that are associated with vernalization and freezing tolerance (FT) in wheat. The level of expression of TaVRT-1 is positively associated with the vernalization response and transition from vegetative to reproductive phase and is negatively associated with the accumulation of COR genes and degree of FT. Comparisons among different wheat genotypes, near-isogenic lines, and cereal species, which differ in their vernalization response and FT, indicated that the gene is inducible only in those species that require vernalization, whereas it is constitutively expressed in spring habit genotypes. In addition, experiments using both the photoperiod sensitive barley (Hordeum vulgare cv Dicktoo) and short or long day de-acclimated wheat revealed that the expression of TaVRT-1 is also regulated by photoperiod. These expression studies indicate that photoperiod and vernalization may regulate this gene through separate pathways. We suggest that TaVRT-1 is a key developmental gene in the regulatory pathway that controls the transition from the vegetative to reproductive phase in cereals. PMID- 12913143 TI - Hyperphosphorylation of a mitochondrial protein, prohibitin, is induced by calyculin A in a rice lesion-mimic mutant cdr1. AB - The rice (Oryza sativa) lesion-mimic mutants, cell death and resistance (cdr), show spontaneous cell death on the entire leaf and exhibited significant resistance to the rice blast fungus. Our previous studies showed that CDR1 and CDR2 genes negatively regulated the phosphorylation steps leading to the activation of NADPH oxidase, which is associated with oxidative burst. To identify novel factors involved in the phosphorylation steps, the phosphorylation level of total proteins was compared between cdr mutants and wild type using two dimensional gel electrophoresis. Here, we show that the phosphorylation level of four proteins in cdr1 was increased as compared with the wild type after calyculin A treatment. Partial amino acid sequences revealed that one of the four proteins is homologous to prohibitin (PHB), which has been shown to be associated with senescence and cell death and to function as a chaperone in the assembly of mitochondrial respiratory chain complex in yeast and mammals. Analysis of green fluorescent protein fusions indicated that rice PHB (OsPHB1) was targeted to mitochondria as found in yeast and mammals, suggesting a possibility that PHB is involved in defense response and/or programmed cell death through the mitochondrial function. PMID- 12913144 TI - MPB2C, a microtubule-associated plant protein binds to and interferes with cell to-cell transport of tobacco mosaic virus movement protein. AB - The movement protein of tobacco mosaic virus, MP30, mediates viral cell-to-cell transport via plasmodesmata. The complex MP30 intra- and intercellular distribution pattern includes localization to the endoplasmic reticulum, cytoplasmic bodies, microtubules, and plasmodesmata and likely requires interaction with plant endogenous factors. We have identified and analyzed an MP30-interacting protein, MPB2C, from the host plant Nicotiana tabacum. MPB2C constitutes a previously uncharacterized microtubule-associated protein that binds to and colocalizes with MP30 at microtubules. In vivo studies indicate that MPB2C mediates accumulation of MP30 at microtubules and interferes with MP30 cell to-cell movement. In contrast, intercellular transport of a functionally enhanced MP30 mutant, which does not accumulate and colocalize with MP30 at microtubules, is not impaired by MPB2C. Together, these data support the concept that MPB2C is not required for MP30 cell-to-cell movement but may act as a negative effector of MP30 cell-to-cell transport activity. PMID- 12913145 TI - Expression of U1 small nuclear ribonucleoprotein 70K antisense transcript using APETALA3 promoter suppresses the development of sepals and petals. AB - U1 small nuclear ribonucleoprotein (snRNP)-70K (U1-70K), a U1 snRNP-specific protein, is involved in the early stages of spliceosome formation. In non-plant systems, it is involved in constitutive and alternative splicing. It has been shown that U1snRNP is dispensable for in vitro splicing of some animal pre-mRNAs, and inactivation of U1-70K in yeast (Saccharomyces cerevisiae) is not lethal. As in yeast and humans (Homo sapiens), plant U1-70K is coded by a single gene. In this study, we blocked the expression of Arabidopsis U1-70K in petals and stamens by expressing U1-70K antisense transcript using the AP3 (APETALA3) promoter specific to these floral organs. Flowers of transgenic Arabidopsis plants expressing U1-70K antisense transcript showed partially developed stamens and petals that are arrested at different stages of development. In some transgenic lines, flowers have rudimentary petals and stamens and are male sterile. The severity of the phenotype is correlated with the level of the antisense transcript. Molecular analysis of transgenic plants has confirmed that the observed phenotype is not due to disruption of whorl-specific homeotic genes, AP3 or PISTILLATA, responsible for petal and stamen development. The AP3 transcript was not detected in transgenic flowers with severe phenotype. Flowers of Arabidopsis plants transformed with a reporter gene driven by the same promoter showed no abnormalities. These results show that U1-70K is necessary for the development of sepals and petals and is an essential gene in plants. PMID- 12913146 TI - C-terminal KDEL sequence of a KDEL-tailed cysteine proteinase (sulfhydryl endopeptidase) is involved in formation of KDEL vesicle and in efficient vacuolar transport of sulfhydryl-endopeptidase. AB - Sulfhydryl-endopeptidase (SH-EP) is a papain-type vacuolar proteinase expressed in cotyledons of germinated Vigna mungo seeds, and the enzyme possesses a C terminal propeptide containing KDEL tail, an endoplasmic reticulum retention signal for soluble proteins. SH-EP is transported to vacuoles via a KDEL vesicle (KV) through a Golgi complex-independent route. To see the function of the KDEL sequence of SH-EP, wild-type SH-EP and its KDEL deletion mutant (SH-EPDeltaKDEL) were heterologously expressed in Arabidopsis and in cultured tobacco Bright Yellow 2 cells, and their intracellular transport pathways and localizations were analyzed. A combination of the results from analyses for transformed Arabidopsis and tobacco (Nicotiana tabacum) cells indicated that wild-type SH-EP is packed into KV-like vesicles through the KDEL sequence and is transported to vacuoles in the cells of transformants. In contrast, KV was not formed/induced in the cells expressing SH-EPDeltaKDEL, and the mutant protein was mainly secreted. Therefore, the C-terminal KDEL sequence of the KDEL-tailed cysteine proteinase is thought to be involved in the formation of KV, and in the efficient vacuolar transport of the proteins through KV. PMID- 12913147 TI - Overexpression of the disease resistance gene Pto in tomato induces gene expression changes similar to immune responses in human and fruitfly. AB - The Pto gene encodes a serine/threonine protein kinase that confers resistance in tomato (Lycopersicon esculentum) to Pseudomonas syringae pv tomato strains that express the type III effector protein AvrPto. Constitutive overexpression of Pto in tomato, in the absence of AvrPto, activates defense responses and confers resistance to several diverse bacterial and fungal plant pathogens. We have used a series of gene discovery and expression profiling methods to examine the effect of Pto overexpression in tomato leaves. Analysis of the tomato expressed sequence tag database and suppression subtractive hybridization identified 600 genes that were potentially differentially expressed in Pto-overexpressing tomato plants compared with a sibling line lacking Pto. By using cDNA microarrays, we verified changes in expression of many of these genes at various time points after inoculation with P. syringae pv tomato (avrPto) of the resistant Pto overexpressing line and the susceptible sibling line. The combination of these three approaches led to the identification of 223 POR (Pto overexpression responsive) genes. Strikingly, 40% of the genes induced in the Pto-overexpressing plants previously have been shown to be differentially expressed during the human (Homo sapiens) and/or fruitfly (Drosophila melanogaster) immune responses. PMID- 12913148 TI - PAUSED, a putative exportin-t, acts pleiotropically in Arabidopsis development but is dispensable for viability. AB - Exportin-t was first identified in humans as a protein that mediates the export of tRNAs from the nucleus to the cytoplasm. Mutations in Los1p, the Saccharomyces cerevisiae exportin-t homolog, result in nuclear accumulation of tRNAs. Because no exportin-t mutants have been reported in multicellular organisms, the developmental functions of exportin-t have not been determined. Here, we report the isolation and characterization of two Arabidopsis exportin-t mutants, paused 5 and paused-6. The mutant phenotypes indicate that exportin-t acts pleiotropically in plant development. In particular, paused-5 and paused-6 result in delayed leaf formation during vegetative development. The two paused mutations also cause the transformation of reproductive organs into perianth organs in the hua1-1 hua2-1 background, which is partially defective in reproductive organ identity specification. The floral phenotypes of hua1-1 hua2-1 paused mutants resemble those of mutations in the floral homeotic gene AGAMOUS. Moreover, paused 5 enhances the mutant phenotypes of two floral meristem identity genes, LEAFY and APETALA1. The developmental defects caused by paused mutations confirm the important roles of exportin-t in gene expression in multicellular organisms. In addition, a paused null allele, paused-6, is still viable, suggesting the presence of redundant tRNA export pathway(s) in Arabidopsis. PMID- 12913150 TI - Structure and properties of an engineered transketolase from maize. AB - The gene specifying plastid transketolase (TK) of maize (Zea mays) was cloned from a cDNA library by southern blotting using a heterologous probe from sorghum (Sorghum bicolor). A recombinant fusion protein comprising thioredoxin of Escherichia coli and mature TK of maize was expressed at a high level in E. coli and cleaved with thrombin, affording plastid TK. The protein in complex with thiamine pyrophoshate was crystallized, and its structure was solved by molecular replacement. The enzyme is a C2 symmetric homodimer closely similar to the enzyme from yeast (Saccharomyces cerevisiae). Each subunit is folded into three domains. The two topologically equivalent active sites are located in the subunit interface region and resemble those of the yeast enzyme. PMID- 12913149 TI - Thermotolerant guard cell protoplasts of tree tobacco do not require exogenous hormones to survive in culture and are blocked from reentering the cell cycle at the G1-to-S transition. AB - When guard cell protoplasts (GCPs) of tree tobacco [Nicotiana glauca (Graham)] are cultured at 32 degrees C with an auxin (1-napthaleneacetic acid) and a cytokinin (6-benzylaminopurine), they reenter the cell cycle, dedifferentiate, and divide. GCPs cultured similarly but at 38 degrees C and with 0.1 micro M +/- cis,trans-abscisic acid (ABA) remain differentiated. GCPs cultured at 38 degrees C without ABA dedifferentiate partially but do not divide. Cell survival after 1 week is 70% to 80% under all of these conditions. In this study, we show that GCPs cultured for 12 to 24 h at 38 degrees C accumulate heat shock protein 70 and develop a thermotolerance that, upon transfer of cells to 32 degrees C, enhances cell survival but inhibits cell cycle reentry, dedifferentiation, and division. GCPs dedifferentiating at 32 degrees C require both 1-napthaleneacetic acid and 6 benzylaminopurine to survive, but thermotolerant GCPs cultured at 38 degrees C +/ ABA do not require either hormone for survival. Pulse-labeling experiments using 5-bromo-2-deoxyuridine indicate that culture at 38 degrees C +/- ABA prevents dedifferentiation of GCPs by blocking cell cycle reentry at G1/S. Cell cycle reentry at 32 degrees C is accompanied by loss of a 41-kD polypeptide that cross reacts with antibodies to rat (Rattus norvegicus) extracellular signal-regulated kinase 1; thermotolerant GCPs retain this polypeptide. A number of polypeptides unique to thermotolerant cells have been uncovered by Boolean analysis of two dimensional gels and are targets for further analysis. GCPs of tree tobacco can be isolated in sufficient numbers and with the purity required to study plant cell thermotolerance and its relationship to plant cell survival, growth, dedifferentiation, and division in vitro. PMID- 12913151 TI - Peptide and amino acid transporters are differentially regulated during seed development and germination in faba bean. AB - Two peptide transporter (PTR) homologs have been isolated from developing seeds of faba bean (Vicia faba). VfPTR1 was shown to be a functional peptide transporter through complementation of a yeast mutant. Expression patterns of VfPTR1 and VfPTR2 as well as of the amino acid permease VfAAP1 (Miranda et al., 2001) were compared throughout seed development and germination. In developing seeds, the highest levels of VfPTR1 transcripts were reached during midcotyledon development, whereas VfAAP1 transcripts were most abundant during early cotyledon development, before the appearance of storage protein gene transcripts, and were detectable until late cotyledon development. During early germination, VfPTR1 mRNA appeared first in cotyledons and later, during seedling growth, also in axes and roots. Expression of VfPTR2 and VfAAP1 was delayed compared with VfPTR1, and was restricted to the nascent organs of the seedlings. Localization of VfPTR1 transcripts showed that this PTR is temporally and spatially regulated during cotyledon development. In germinating seeds, VfPTR1 mRNA was localized in root hairs and root epidermal cells, suggesting a role in nutrient uptake from the soil. In seedling roots, VfPTR1 was repressed by a dipeptide and by an amino acid, whereas nitrate was without influence. PMID- 12913152 TI - BWMK1, a rice mitogen-activated protein kinase, locates in the nucleus and mediates pathogenesis-related gene expression by activation of a transcription factor. AB - Mitogen-activated protein kinase (MAPK) cascades are known to transduce plant defense signals, but the downstream components of the MAPK have as yet not been elucidated. Here, we report an MAPK from rice (Oryza sativa), BWMK1, and a transcription factor, OsEREBP1, phosphorylated by the kinase. The MAPK carries a TDY phosphorylation motif instead of the more common TEY motif in its kinase domain and has an unusually extended C-terminal domain that is essential to its kinase activity and translocation to the nucleus. The MAPK phosphorylates OsEREBP1 that binds to the GCC box element (AGCCGCC) of the several basic pathogenesis-related gene promoters, which in turn enhances DNA-binding activity of the factor to the cis element in vitro. Transient co-expression of the BWMK1 and OsEREBP1 in Arabidopsis protoplasts elevates the expression of the beta glucuronidase reporter gene driven by the GCC box element. Furthermore, transgenic tobacco (Nicotiana tabacum) plants overexpressing BWMK1 expressed many pathogenesis-related genes at higher levels than wild-type plants with an enhanced resistance to pathogens. These findings suggest that MAPKs contribute to plant defense signal transduction by phosphorylating one or more transcription factors. PMID- 12913153 TI - Induction of hypersensitive cell death by hydrogen peroxide produced through polyamine degradation in tobacco plants. AB - Screening immediate-early responding genes during the hypersensitive response (HR) against tobacco mosaic virus infection in tobacco (Nicotiana tabacum) plants, we identified a gene encoding ornithine decarboxylase. Subsequent analyses showed that other genes involved in polyamine biosynthesis were also up regulated, resulting in the accumulation of polyamines in apoplasts of tobacco mosaic virus-infected leaves. Inhibitors of polyamine biosynthesis, alpha difluoromethyl-ornithine, however, suppressed accumulation of polyamines, and the rate of HR was reduced. In contrast, polyamine infiltration into a healthy leaf induced the generation of hydrogen peroxide and simultaneously caused HR-like cell death. Polyamine oxidase activity in the apoplast increased up to 3-fold that of the basal level during the HR, and its suppression with a specific inhibitor, guazatine, resulted in reduced HR. Because it is established that hydrogen peroxide is one of the degradation products of polyamines, these results indicate that one of the biochemical events in the HR is production of polyamines, whose degradation induces hydrogen peroxide, eventually resulting in hypersensitive cell death. PMID- 12913154 TI - Nod factor-induced root hair curling: continuous polar growth towards the point of nod factor application. AB - A critical step in establishing a successful nitrogen-fixing symbiosis between rhizobia and legume plants is the entrapment of the bacteria between root hair cell walls, usually in characteristic 180 degrees to 360 degrees curls, shepherd's crooks, which are formed by the host's root hairs. Purified bacterial signal molecules, the nodulation factors (NFs), which are lipochitooligosaccharides, induce root hair deformation in the appropriate host legume and have been proposed to be a key player in eliciting root hair curling. However, for curling to occur, the presence of intact bacteria is thought to be essential. Here, we show that, when spot applied to one side of the growing Medicago truncatula root hair tip, purified NF alone is sufficient to induce reorientation of the root hair growth direction, or a full curl. Using wild-type M. truncatula containing the pMtENOD11::GUS construct, we demonstrate that MtENOD11::GUS is expressed after spot application. The data have been incorporated into a cell biological model, which explains the formation of shepherd's crook curls around NF-secreting rhizobia by continuous tip growth reorientation. PMID- 12913155 TI - Three nicotianamine synthase genes isolated from maize are differentially regulated by iron nutritional status. AB - Nicotianamine synthase (NAS) is an enzyme that is critical for the biosynthesis of the mugineic acid family of phytosiderophores in graminaceous plants, and for the homeostasis of metal ions in nongraminaceous plants. We isolated one genomic NAS clone, ZmNAS3, and two cDNA NAS clones, ZmNAS1 and ZmNAS2, from maize (Zea mays cv Alice). In agreement with the increased secretion of phytosiderophores with Fe deficiency, ZmNAS1 and ZmNAS2 were positively expressed only in Fe deficient roots. In contrast, ZmNAS3 was expressed under Fe-sufficient conditions, and was negatively regulated by Fe deficiency. This is the first report describing down-regulation of NAS gene expression in response to Fe deficiency in plants, shedding light on the role of nicotianamine in graminaceous plants, other than as a precursor in phytosiderophore production. ZmNAS1-green fluorescent protein (sGFP) and ZmNAS2-sGFP were localized at spots in the cytoplasm of onion (Allium cepa) epidermal cells, whereas ZmNAS3-sGFP was distributed throughout the cytoplasm of these cells. ZmNAS1 and ZmNAS3 showed NAS activity in vitro, whereas ZmNAS2 showed none. Due to its duplicated structure, ZmNAS2 was much larger (65.8 kD) than ZmNAS1, ZmNAS3, and previously characterized NAS proteins (30-38 kD) from other plant species. We reveal that maize has two types of NAS proteins based on their expression pattern and subcellular localization. PMID- 12913156 TI - Expression profiling of cytokinin action in Arabidopsis. AB - The phytohormone cytokinin is an important regulator of plant growth and development; however, relatively few genes that mediate cytokinin responses have been identified. Genome-wide analyses of Arabidopsis seedlings using the approximately 8,300-element Affymetrix Arabidopsis GeneChips (Affymetrix, Santa Clara, CA) to examine cytokinin-responsive genes were conducted, revealing at least 30 genes whose steady-state level of mRNA was elevated and at least 40 that were down-regulated at multiple time points after application of cytokinin. The cytokinin up-regulated genes include the type-A Arabidopsis response regulators (ARRs), which had been shown previously to be cytokinin primary response genes, cytokinin oxidase, which encodes an enzyme that degrades cytokinins, and several transcription factors. Cytokinin down-regulated genes include several peroxidases and kinases and an E3 ubiquitin ligase. We identified a common sequence motif enriched in the upstream regions of the most consistently cytokinin up-regulated genes. This motif is highly similar to the optimal DNA-binding sites for ARR1/ARR2, type-B ARRs that have been implicated in the transcriptional elevation of the type-A ARRs. Additionally, genome-wide analyses of cytokinin receptor mutants (wol/cre1) revealed large-scale changes in gene expression, including down-regulation of the type-A ARRs and several meristem and cell cycle genes, such as CycD3. Mutations in CRE1 reduced but did not eliminate the effect of cytokinin on gene expression for a subset of cytokinin-responsive genes and had little or no effect on others, suggesting functional redundancy among the cytokinin receptors. PMID- 12913157 TI - Constitutive E2F expression in tobacco plants exhibits altered cell cycle control and morphological change in a cell type-specific manner. AB - The E2F family plays a pivotal role in cell cycle control and is conserved among plants and animals, but not in fungi. This provides for the possibility that the E2F family was integrated during the development of higher organisms, but little is known about this. We examined the effect of E2F ectopically expressed in transgenic tobacco (Nicotiana tabacum) plants on growth and development using E2Fa (AtE2F3) and DPa from Arabidopsis. E2Fa-DPa double transgenic lines exhibited altered phenotypes with curled leaves, round shaped petals, and shortened pistils. In mature but not immature leaves of the double transgenic lines, there were enlarged nuclei with increasing ploidy levels accompanied by the ectopic expression of S phase- but not M phase-specific genes. This indicates that a high expression of E2F promotes endoreduplication by accelerating S phase entry in terminally differentiated cells with limited mitotic activity. Furthermore, mature leaves of the transgenic plants contained increased numbers of small cells, especially on the palisade (adaxial) side of the outer region toward the edge, and the leaf strips exhibited hormone-independent callus formation when cultured in vitro. These observations suggest that an enhanced E2F activity modulates cell cycle in a cell type-specific manner and affects plant morphology depending on a balance between activities for committing to S phase and M phase, which likely differ between organs or tissues. PMID- 12913158 TI - The Arabidopsis NHL3 gene encodes a plasma membrane protein and its overexpression correlates with increased resistance to Pseudomonas syringae pv. tomato DC3000. AB - The Arabidopsis genome contains a family of NDR1/HIN1-like (NHL) genes that show homology to the nonrace-specific disease resistance (NDR1) and the tobacco (Nicotiana tabacum) harpin-induced (HIN1) genes. NHL3 is a pathogen-responsive member of this NHL gene family that is potentially involved in defense. In independent transgenic NHL3-overexpressing plant lines, a clear correlation between increased resistance to virulent Pseudomonas syringae pv. tomato DC3000 and enhanced NHL3 transcript levels was seen. These transgenic plants did not show enhanced pathogenesis-related gene expression or reactive oxygen species accumulation. Biochemical and localization experiments were performed to assist elucidation of how NHL3 may confer enhanced disease resistance. Gene constructs expressing amino-terminal c-myc-tagged or carboxyl-terminal hemagglutinin epitope (HA)-tagged NHL3 demonstrated membrane localization in transiently transformed tobacco leaves. Stable Arabidopsis transformants containing the NHL3-HA construct corroborated the findings observed in tobacco. The detected immunoreactive proteins were 10 kD larger than the calculated size and could be partially accounted for by the glycosylation state. However, the expected size was not attained with deglycosylation, suggesting possibly additional posttranslational modification. Detergent treatment, but not chemicals used to strip membrane associated proteins, could displace the immunoreactive signal from microsomal fractions, showing that NHL3 is tightly membrane associated. Furthermore, immunofluorescence and immunogold labeling, coupled with two-phase partitioning techniques, revealed plasma membrane localization of NHL3-HA. This subcellular localization of NHL3 positions it at an initial contact site to pathogens and may be important in facilitating interception of pathogen-derived signals. PMID- 12913159 TI - The putative Arabidopsis arp2/3 complex controls leaf cell morphogenesis. AB - The evolutionarily conserved Arp2/3 complex has been shown to activate actin nucleation and branching in several eukaryotes, but its biological functions are not well understood in multicellular organisms. The model plant Arabidopsis provides many advantages for genetic dissection of the function of this conserved actin-nucleating machinery, yet the existence of this complex in plants has not been determined. We have identified Arabidopsis genes encoding homologs of all of the seven Arp2/3 subunits. The function of the putative Arabidopsis Arp2/3 complex has been studied using four homozygous T-DNA insertion mutants for ARP2, ARP3, and ARPC5/p16. All four mutants display identical defects in the development of jigsaw-shaped epidermal pavement cells and branched trichomes in the leaf. These loss-of-function mutations cause mislocalization of diffuse cortical F-actin to the neck region and inhibit lobe extension in pavement cells. The mutant trichomes resemble those treated with the actin-depolymerizing drug cytochalasin D, exhibiting stunted branches but dramatically enlarged stalks due to depolarized growth suggesting defects in the formation of a fine actin network. Our data demonstrate that the putative Arabidopsis Arp2/3 complex controls cell morphogenesis through its roles in cell polarity establishment and polar cell expansion. Furthermore, our data suggest a novel function for the putative Arp2/3 complex in the modulation of the spatial distribution of cortical F-actin and provide evidence that the putative Arp2/3 complex may activate the polymerization of some types of actin filaments in specific cell types. PMID- 12913160 TI - Resemblance and dissemblance of Arabidopsis type II peroxiredoxins: similar sequences for divergent gene expression, protein localization, and activity. AB - The Arabidopsis type II peroxiredoxin (PRXII) family is composed of six different genes, five of which are expressed. On the basis of the nucleotide and protein sequences, we were able to define three subgroups among the PRXII family. The first subgroup is composed of AtPRXII-B, -C, and -D, which are highly similar and localized in the cytosol. AtPRXII-B is ubiquitously expressed. More striking is the specific expression of AtPRXII-C and AtPRXII-D localized in pollen. The second subgroup comprises the mitochondrial AtPRXII-F, the corresponding gene of which is expressed constitutively. We show that AtPRXII-E, belonging to the last subgroup, is expressed mostly in reproductive tissues and that its product is addressed to the plastid. By in vitro enzymatic experiments, we demonstrate that glutaredoxin is the electron donor of recombinant AtPRXII-B for peroxidase reaction, but the donors of AtPRXII-E and AtPRXII-F have still to be identified. PMID- 12913161 TI - A bypass of sucrose synthase leads to low internal oxygen and impaired metabolic performance in growing potato tubers. AB - Plants possess two alternative biochemical pathways for sucrose (Suc) degradation. One involves hydrolysis by invertase followed by phosphorylation via hexokinase and fructokinase, and the other route-which is unique to plants involves a UDP-dependent cleavage of Suc that is catalyzed by Suc synthase (SuSy). In the present work, we tested directly whether a bypass of the endogenous SuSy route by ectopic overexpression of invertase or Suc phosphorylase affects internal oxygen levels in growing tubers and whether this is responsible for their decreased starch content. (a) Oxygen tensions were lower within transgenic tubers than in wild-type tubers. Oxygen tensions decreased within the first 10 mm of tuber tissue, and this gradient was steeper in transgenic tubers. (b) Invertase-overexpressing tubers had higher activities of glyceraldehyde-3 phosphate dehydrogenase, lactate dehydrogenase, and alcohol dehydrogenase, and (c) higher levels of lactate. (d) Expression of a low-oxygen-sensitive Adh1-beta glucuronidase reporter gene construct was more strongly induced in the invertase overexpressing background compared with wild-type background. (e) Intact transgenic tubers had lower ATP to ADP ratios than the wild type. ATP to ADP ratio was restored to wild type, when discs of transgenic tubers were incubated at 21% (v/v) oxygen. (f) Starch decreased from the periphery to the center of the tuber. This decrease was much steeper in the transgenic lines, leading to lower starch content especially near the center of the tuber. (g) Metabolic fluxes (based on redistribution of (14)C-glucose) and ATP to ADP ratios were analyzed in more detail, comparing discs incubated at various external oxygen tensions (0%, 1%, 4%, 8%, 12%, and 21% [v/v]) with intact tubers. Discs of Suc phosphorylase expressing lines had similar ATP to ADP ratios and made starch as fast as wild type in high oxygen but had lower ATP to ADP ratios and lower rates of starch synthesis than wild type at low-oxygen tensions typical to those found inside an intact tuber. (h) In discs of wild-type tubers, subambient oxygen concentrations led to a selective increase in the mRNA levels of specific SuSy genes, whereas the mRNA levels of genes encoding vacuolar and apoplastic invertases decreased. (i) These results imply that repression of invertase and mobilization of Suc via the energetically less costly route provided by SuSy is important in growing tubers because it conserves oxygen and allows higher internal oxygen tensions to be maintained than would otherwise be possible. PMID- 12913162 TI - Purification and characterization of four beta-expansins (Zea m 1 isoforms) from maize pollen. AB - Four proteins with wall extension activity on grass cell walls were purified from maize (Zea mays) pollen by conventional column chromatography and high performance liquid chromatography. Each is a basic glycoprotein (isoelectric point = 9.1-9.5) of approximately 28 kD and was identified by immunoblot analysis as an isoform of Zea m 1, the major group 1 allergen of maize pollen and member of the beta-expansin family. Four distinctive cDNAs for Zea m 1 were identified by cDNA library screening and by GenBank analysis. One pair (GenBank accession nos. AY104999 and AY104125) was much closer in sequence to well-characterized allergens such as Lol p 1 and Phl p 1 from ryegrass (Lolium perenne) and Phleum pretense, whereas a second pair was much more divergent. The N-terminal sequence and mass spectrometry fingerprint of the most abundant isoform (Zea m 1d) matched that predicted for AY197353, whereas N-terminal sequences of the other isoforms matched or nearly matched AY104999 and AY104125. Highly purified Zea m 1d induced extension of a variety of grass walls but not dicot walls. Wall extension activity of Zea m 1d was biphasic with respect to protein concentration, had a broad pH optimum between 5 and 6, required more than 50 micro g mL(-1) for high activity, and led to cell wall breakage after only approximately 10% extension. These characteristics differ from those of alpha-expansins. Some of the distinctive properties of Zea m 1 may not be typical of beta-expansins as a class but may relate to the specialized function of this beta-expansin in pollen function. PMID- 12913163 TI - Regulated phosphorylation of 40S ribosomal protein S6 in root tips of maize. AB - Ribosomal protein S6 (RPS6) is located in the mRNA binding site of the 40S subunit of cytosolic ribosomes. Two maize (Zea mays) rps6 genes were identified that encode polypeptides (30 kD, 11.4 pI) with strong primary amino acid sequence and predicted secondary structure similarity to RPS6 of other eukaryotes. Maize RPS6 was analyzed by the use of two-dimensional gel electrophoresis systems, in vivo labeling with [(32)P]P(i) and immunological detection. Nine RPS6 isoforms were resolved in a two-dimensional basic-urea/sodium dodecyl sulfate polyacrylamide gel electrophoresis system. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry performed on trypsin-digested isoforms identified four serine (Ser) and one threonine (Thr) residue in the carboxy-terminal region as phosphorylation sites (RRS(238)KLS(241)AAAKAS(247)AAT(250)S(251)A-COOH). Heterogeneity in RPS6 phosphorylation was a consequence of the presence of zero to five phosphorylated residues. Phosphorylated isoforms fell into two groups characterized by (a) sequential phosphorylation of Ser-238 and Ser-241 and (b) the absence of phospho Ser-238 and presence of phospho-Ser-241. The accumulation of hyper-phosphorylated isoforms with phospho-Ser-238 was reduced in response to oxygen deprivation and heat shock, whereas accumulation of these isoforms was elevated by cold stress. Salt and osmotic stress had no reproducible effect on RPS6 phosphorylation. The reduction in hyper-phosphorylated isoforms under oxygen deprivation was blocked by okadaic acid, a Ser/Thr phosphatase inhibitor. By contrast, the recovery of hyper-phosphorylated isoforms upon re-oxygenation was blocked by LY-294002, an inhibitor of phosphatidylinositol 3-kinases. Thus, differential activity of phosphatase(s) and kinase(s) determine complex heterogeneity in RPS6 phosphorylation. PMID- 12913164 TI - Salt stress activation of wound-related genes in tomato plants. AB - Plants respond to various stresses by expressing distinct sets of genes. The effects of multiple stresses on plants and their interactions are not well understood. We have discovered that salt stress causes the accumulation of proteinase inhibitors and the activation of other wound-related genes in tomato (Lycopersicon esculentum) plants. Salt stress was also found to enhance the plant's response to wounding locally and systemically. The tomato mutant (def-1), which has an impairment in the octadecanoid pathway, displayed a severe reduction in the accumulation of proteinase inhibitors under salt stress, indicating that salt stress-induced accumulation of proteinase inhibitors was jasmonic acid dependent. The analysis of salt stress in another tomato mutant, spr-1, which carries a mutation in a systemin-specific signaling component, and transgenic tomato plants that express an antisense-prosystemin cDNA, showed that prosystemin activity was not required for the salt-induced accumulation of proteinase inhibitors, but was necessary to achieve maximal levels. These results suggest that a prosystemin independent- but jasmonic acid-dependent pathway is utilized for proteinase inhibitor accumulation in response to salt stress. PMID- 12913165 TI - Pea DNA topoisomerase I is phosphorylated and stimulated by casein kinase 2 and protein kinase C. AB - DNA topoisomerase I catalyzes the relaxation of superhelical DNA tension and is vital for DNA metabolism; therefore, it is essential for growth and development of plants. Here, we have studied the phosphorylation-dependent regulation of topoisomerase I from pea (Pisum sativum). The purified enzyme did not show autophosphorylation but was phosphorylated in an Mg(2+)-dependent manner by endogenous protein kinases present in pea nuclear extracts. This phosphorylation was abolished with calf intestinal alkaline phosphatase and lambda phosphatase. It was also phosphorylated by exogenous casein kinase 2 (CK2), protein kinase C (PKC; from animal sources), and an endogenous pea protein, which was purified using a novel phorbol myristate acetate affinity chromatography method. All of these phosphorylations were inhibited by heparin (inhibitor of CK2) and calphostin (inhibitor of PKC), suggesting that pea topoisomerase I is a bona fide substrate for these kinases. Spermine and spermidine had no effect on the CK2 mediated phosphorylation, suggesting that it is polyamine independent. Phospho amino acid analysis showed that only serine residues were phosphorylated, which was further confirmed using antiphosphoserine antibody. The topoisomerase I activity increased after phosphorylation with exogenous CK2 and PKC. This study shows that these kinases may contribute to the physiological regulation of DNA topoisomerase I activity and overall DNA metabolism in plants. PMID- 12913166 TI - Thylakoid-bound ascorbate peroxidase mutant exhibits impaired electron transport and photosynthetic activity. AB - In chloroplasts, stromal and thylakoid-bound ascorbate peroxidases (tAPX) play a major role in the removal of H(2)O(2) produced during photosynthesis. Here, we report that hexaploid wheat (Triticum aestivum) expresses three homeologous tAPX genes (TaAPX-6A, TaAPX-6B, and TaAPX-6D) mapping on group-6 chromosomes. The tAPX activity of a mutant line lacking TaAPX-6B was 40% lower than that of the wild type. When grown at high-light intensity photosystem II electron transfer, photosynthetic activity and biomass accumulation were significantly reduced in this mutant, suggesting that tAPX activity is essential for photosynthesis. Despite the reduced tAPX activity, mutant plants did not exhibit oxidative damage probably due to the reduced photochemical activity. This might be the result of a compensating mechanism to prevent oxidative damage having as a consequence a decrease in growth of the tAPX mutant plants. PMID- 12913167 TI - An anaplerotic role for mitochondrial carbonic anhydrase in Chlamydomonas reinhardtii. AB - Previous studies of the mitochondrial carbonic anhydrase (mtCA) of Chlamydomonas reinhardtii showed that expression of the two genes encoding this enzyme activity required photosynthetically active radiation and a low CO(2) concentration. These studies suggested that the mtCA was involved in the inorganic carbon concentrating mechanism. We have now shown that the expression of the mtCA at low CO(2) concentrations decreases when the external NH(4)(+) concentration decreases, to the point of being undetectable when NH(4)(+) supply restricts the rate of photoautotrophic growth. The expression of mtCA can also be induced at supra-atmospheric partial pressure of CO(2) by increasing the NH(4)(+) concentration in the growth medium. Conditions that favor mtCA expression usually also stimulate anaplerosis. We therefore propose that the mtCA is involved in supplying HCO(3)(-) for anaplerotic assimilation catalyzed by phosphoenolpyruvate carboxylase, which provides C skeletons for N assimilation under some circumstances. PMID- 12913168 TI - PAUSED encodes the Arabidopsis exportin-t ortholog. AB - Los1p/exportin-t (XPOT) mediates the nuclear export of tRNAs in yeast and mammals. The requirements for this transport pathway are unclear, however, because los1 mutations do not affect yeast growth, and the phenotype of XPOT mutations in mammals is unknown. Here, we show that PAUSED (PSD) is the Arabidopsis ortholog of LOS1/XPOT and is capable of rescuing the tRNA export defect of los1 in Brewer's yeast (Saccharomyces cerevisiae), suggesting that its function has been conserved. Putative null alleles of PSD disrupt the initiation of the shoot apical meristem and delay leaf initiation after germination, the emergence of the radicle and lateral roots, and the transition to flowering. Plants doubly mutant for psd and hasty, the Arabidopsis ortholog of exportin 5, are viable but have a more severe phenotype than either single mutant. These results suggest that PSD plays a role in tRNA export in Arabidopsis, but that at least one-and perhaps several-additional tRNA export pathways also exist. The PSD transcript is broadly expressed during development and is alternatively spliced in the 3'-untranslated region. No temporal or spatial difference in the abundance of different splice forms was observed. We propose that the mutant phenotype of psd reflects defects in developmental events and cell/tissue types that require elevated levels of protein synthesis and are therefore acutely sensitive to a reduction in tRNA export. PMID- 12913169 TI - Changes in the redox potential of primary and secondary electron-accepting quinones in photosystem II confer increased resistance to photoinhibition in low temperature-acclimated Arabidopsis. AB - Exposure of control (non-hardened) Arabidopsis leaves for 2 h at high irradiance at 5 degrees C resulted in a 55% decrease in photosystem II (PSII) photochemical efficiency as indicated by F(v)/F(m). In contrast, cold-acclimated leaves exposed to the same conditions showed only a 22% decrease in F(v)/F(m). Thermoluminescence was used to assess the possible role(s) of PSII recombination events in this differential resistance to photoinhibition. Thermoluminescence measurements of PSII revealed that S(2)Q(A)(-) recombination was shifted to higher temperatures, whereas the characteristic temperature of the S(2)Q(B)(-) recombination was shifted to lower temperatures in cold-acclimated plants. These shifts in recombination temperatures indicate higher activation energy for the S(2)Q(A)(-) redox pair and lower activation energy for the S(2)Q(B)(-) redox pair. This results in an increase in the free-energy gap between P680(+)Q(A)(-) and P680(+)Pheo(-) and a narrowing of the free energy gap between primary and secondary electron-accepting quinones in PSII electron acceptors. We propose that these effects result in an increased population of reduced primary electron accepting quinone in PSII, facilitating non-radiative P680(+)Q(A)(-) radical pair recombination. Enhanced reaction center quenching was confirmed using in vivo chlorophyll fluorescence-quenching analysis. The enhanced dissipation of excess light energy within the reaction center of PSII, in part, accounts for the observed increase in resistance to high-light stress in cold-acclimated Arabidopsis plants. PMID- 12913170 TI - Systematic trans-genomic comparison of protein kinases between Arabidopsis and Saccharomyces cerevisiae. AB - The genome of the budding yeast (Saccharomyces cerevisiae) provides an important paradigm for transgenomic comparisons with other eukaryotic species. Here, we report a systematic comparison of the protein kinases of yeast (119 kinases) and a reference plant Arabidopsis (1,019 kinases). Using a whole-protein-based, hierarchical clustering approach, the complete set of protein kinases from both species were clustered. We validated our clustering by three observations: (a) clustering pattern of functional orthologs proven in genetic complementation experiments, (b) consistency with reported classifications of yeast kinases, and (c) consistency with the biochemical properties of those Arabidopsis kinases already experimentally characterized. The clustering pattern identified no overlap between yeast kinases and the receptor-like kinases (RLKs) of Arabidopsis. Ten more kinase families were found to be specific for one of the two species. Among them, the calcium-dependent protein kinase and phosphoenolpyruvate carboxylase kinase families are specific for plants, whereas the Ca(2+)/calmodulin-dependent protein kinase and provirus insertion in mouse like kinase families were found only in yeast and animals. Three yeast kinase families, nitrogen permease reactivator/halotolerance-5), polyamine transport kinase, and negative regulator of sexual conjugation and meiosis, are absent in both plants and animals. The majority of yeast kinase families (21 of 26) display Arabidopsis counterparts, and all are mapped into Arabidopsis families of intracellular kinases that are not related to RLKs. Representatives from 11 of the common families (54 kinases from Arabidopsis and 17 from yeast) share an extremely high degree of similarity (blast E value < 10(-80)), suggesting the likelihood of orthologous functions. Selective expansion of yeast kinase families was observed in Arabidopsis. This is most evident for yeast genes CBK1, HRR25, and SNF1 and the kinase family S6K. Reduction of kinase families was also observed, as in the case of the NEK-like family. The distinguishing features between the two sets of kinases are the selective expansion of yeast families and the generation of a limited number of new kinase families for new functionality in Arabidopsis, most notably, the Arabidopsis RLKs that constitute important components of plant intercellular communication apparatus. PMID- 12913171 TI - Stomatal closure during leaf dehydration, correlation with other leaf physiological traits. AB - The question as to what triggers stomatal closure during leaf desiccation remains controversial. This paper examines characteristics of the vascular and photosynthetic functions of the leaf to determine which responds most similarly to stomata during desiccation. Leaf hydraulic conductance (K(leaf)) was measured from the relaxation kinetics of leaf water potential (Psi(l)), and a novel application of this technique allowed the response of K(leaf) to Psi(l) to be determined. These "vulnerability curves" show that K(leaf) is highly sensitive to Psi(l) and that the response of stomatal conductance to Psi(l) is closely correlated with the response of K(leaf) to Psi(l). The turgor loss point of leaves was also correlated with K(leaf) and stomatal closure, whereas the decline in PSII quantum yield during leaf drying occurred at a lower Psi(l) than stomatal closure. These results indicate that stomatal closure is primarily coordinated with K(leaf). However, the close proximity of Psi(l) at initial stomatal closure and initial loss of K(leaf) suggest that partial loss of K(leaf) might occur regularly, presumably necessitating repair of embolisms. PMID- 12913172 TI - Overproduction of cytokinins in petunia flowers transformed with P(SAG12)-IPT delays corolla senescence and decreases sensitivity to ethylene. AB - Plant senescence is regulated by a coordinated genetic program mediated in part by changes in ethylene, abscisic acid (ABA), and cytokinin content. Transgenic plants with delayed senescence are useful for studying interactions between these signaling mechanisms. Expression of ipt, a cytokinin biosynthetic gene from Agrobacterium tumefaciens, under the control of the promoter from a senescence associated gene (SAG12) has been one approach used to delay senescence. We transformed petunia (Petunia x hybrida cv V26) with P(SAG12)-IPT. Two independently transformed lines with extended flower longevity (I-1-7-22 and I-3 18-34) were used to study the effects of elevated cytokinin content on ethylene synthesis and sensitivity and ABA accumulation in petunia corollas. Floral senescence in these lines was delayed 6 to 10 d relative to wild-type (WT) flowers. Ipt transcripts increased in abundance after pollination and were accompanied by increased cytokinin accumulation. Endogenous ethylene production was induced by pollination in both WT and IPT corollas, but this increase was delayed in IPT flowers. Flowers from IPT plants were less sensitive to exogenous ethylene and required longer treatment times to induce endogenous ethylene production, corolla senescence, and up-regulation of the senescence-related Cys protease phcp1. Accumulation of ABA, another hormone regulating flower senescence, was significantly greater in WT corollas, confirming that floral senescence was delayed in IPT plants. These results extend our understanding of the hormone interactions that regulate flower senescence and provide a means of increasing flower longevity. PMID- 12913173 TI - Characterization of tocopherol cyclases from higher plants and cyanobacteria. Evolutionary implications for tocopherol synthesis and function. AB - Tocopherols are lipophilic antioxidants synthesized exclusively by photosynthetic organisms and collectively constitute vitamin E, an essential nutrient for both humans and animals. Tocopherol cyclase (TC) catalyzes the conversion of various phytyl quinol pathway intermediates to their corresponding tocopherols through the formation of the chromanol ring. Herein, the molecular and biochemical characterization of TCs from Arabidopsis (VTE1 [VITAMIN E 1]), Zea mays (SXD1 [Sucrose Export Deficient 1]) and Synechocystis sp. PCC6803 (slr1737) are described. Mutations in the VTE1, SXD1, or slr1737 genes resulted in both tocopherol deficiency and the accumulation of 2,3-dimethyl-6-phytyl-1,4 benzoquinone (DMPBQ), a TC substrate. Recombinant SXD1 and VTE1 proteins are able to convert DMPBQ to gamma-tocopherol in vitro. In addition, expression of maize SXD1 in a Synechocystis sp. PCC6803 slr1737 knockout mutant restored tocopherol synthesis, indicating that TC activity is evolutionarily conserved between plants and cyanobacteria. Sequence analysis identified a highly conserved 30-amino acid C-terminal domain in plant TCs that is absent from cyanobacterial orthologs. vte1 2 causes a truncation within this C-terminal domain, and the resulting mutant phenotype suggests that this domain is necessary for TC activity in plants. The defective export of Suc in sxd1 suggests that in addition to presumed antioxidant activities, tocopherols or tocopherol breakdown products also function as signal transduction molecules, or, alternatively, the DMPBQ that accumulates in sxd1 disrupts signaling required for efficient Suc export in maize. PMID- 12913174 TI - Nod factor inhibition of reactive oxygen efflux in a host legume. AB - Hydrogen peroxide (H(2)O(2)) efflux was measured from Medicago truncatula root segments exposed to purified Nod factor and to poly-GalUA (PGA) heptamers. Nod factor, at concentrations > 100 pM, reduced H(2)O(2) efflux rates to 60% of baseline levels beginning 20 to 30 min after exposure, whereas the PGA elicitor, at > 75 nM, caused a rapid increase in H(2)O(2) efflux to >200% of baseline rates. Pretreatment of plants with Nod factor alters the effect of PGA by limiting the maximum H(2)O(2) efflux rate to 125% of that observed for untreated plants. Two Nod factor-related compounds showed no ability to modulate peroxide efflux, and tomato (Lycopersicon esculentum), a nonlegume, showed no response to 1 nM Nod factor. Seven M. truncatula mutants, lacking the ability to make nodules, were tested for Nod factor effects on H(2)O(2) efflux. The nfp mutant was blocked for suppression of peroxide efflux, whereas the dmi1 and dmi2 mutants, previously shown to be blocked for early Nod factor responses, showed a wild-type peroxide efflux modulation. These data demonstrate that exposure to Nod factor suppresses the activity of the reactive oxygen-generating system used for plant defense responses. PMID- 12913175 TI - Modulation of citrate metabolism alters aluminum tolerance in yeast and transgenic canola overexpressing a mitochondrial citrate synthase. AB - Aluminum (Al) toxicity is a major constraint for crop production in acid soils, although crop cultivars vary in their tolerance to Al. We have investigated the potential role of citrate in mediating Al tolerance in Al-sensitive yeast (Saccharomyces cerevisiae; MMYO11) and canola (Brassica napus cv Westar). Yeast disruption mutants defective in genes encoding tricarboxylic acid cycle enzymes, both upstream (citrate synthase [CS]) and downstream (aconitase [ACO] and isocitrate dehydrogenase [IDH]) of citrate, showed altered levels of Al tolerance. A triple mutant of CS (Deltacit123) showed lower levels of citrate accumulation and reduced Al tolerance, whereas Deltaaco1- and Deltaidh12 deficient mutants showed higher accumulation of citrate and increased levels of Al tolerance. Overexpression of a mitochondrial CS (CIT1) in MMYO11 resulted in a 2- to 3-fold increase in citrate levels, and the transformants showed enhanced Al tolerance. A gene for Arabidopsis mitochondrial CS was overexpressed in canola using an Agrobacterium tumefaciens-mediated system. Increased levels of CS gene expression and enhanced CS activity were observed in transgenic lines compared with the wild type. Root growth experiments revealed that transgenic lines have enhanced levels of Al tolerance. The transgenic lines showed enhanced levels of cellular shoot citrate and a 2-fold increase in citrate exudation when exposed to 150 micro M Al. Our work with yeast and transgenic canola clearly suggest that modulation of different enzymes involved in citrate synthesis and turnover (malate dehydrogenase, CS, ACO, and IDH) could be considered as potential targets of gene manipulation to understand the role of citrate metabolism in mediating Al tolerance. PMID- 12913176 TI - Fructans, but not the sucrosyl-galactosides, raffinose and loliose, are affected by drought stress in perennial ryegrass. AB - The aim of this study was to evaluate the putative role of the sucrosyl galactosides, loliose [alpha-D-Gal (1,3) alpha-D-Glc (1,2) beta-D-Fru] and raffinose [alpha-D-Gal (1,6) alpha-D-Glc (1,2) beta-D-Fru], in drought tolerance of perennial ryegrass and to compare it with that of fructans. To that end, the loliose biosynthetic pathway was first established and shown to operate by a UDP Gal: sucrose (Suc) 3-galactosyltransferase, tentatively termed loliose synthase. Drought stress increased neither the concentrations of loliose and raffinose nor the activities of loliose synthase and raffinose synthase (EC 2.4.1.82). Moreover, the concentrations of the raffinose precursors, myoinositol and galactinol, as well as the gene expressions of myoinositol 1-phosphate synthase (EC 5.5.1.4) and galactinol synthase (EC 2.4.1.123) were either decreased or unaffected by drought stress. Taken together, these data are not in favor of an obvious role of sucrosyl-galactosides in drought tolerance of perennial ryegrass at the vegetative stage. By contrast, drought stress caused fructans to accumulate in leaf tissues, mainly in leaf sheaths and elongating leaf bases. This increase was mainly due to the accumulation of long-chain fructans (degree of polymerization > 8) and was not accompanied by a Suc increase. Interestingly, Suc but not fructan concentrations greatly increased in drought-stressed roots. Putative roles of fructans and sucrosyl-galactosides are discussed in relation to the acquisition of stress tolerance. PMID- 12913177 TI - Molecular characterization of a novel lipase-like pathogen-inducible gene family of Arabidopsis. AB - In a differential screening between Arabidopsis plants pretreated with the resistance-inducer beta-aminobutyric acid and untreated control plants, we have identified a gene encoding a novel lipase-like protein, PRLIP1. The abundance of PRLIP1 mRNAs in Arabidopsis leaves was up-regulated by application of beta aminobutyric acid, salicylic acid (SA), and ethylene as well as by various pathogens. Induction of PRLIP1 depended on a functioning SA and ethylene signal transduction pathway but was independent of jasmonate signaling. This novel pathogenesis-related (PR) gene of Arabidopsis belongs to a gene family consisting of six (PRLIP1, PRLIP2, PRLIP4, PRLIP5, PRLIP6, and PRLIP7) closely related members in tandem position on chromosome 5. Among these genes, PRLIP2 also was induced in leaves by SA and infections by pathogens but on a much lower level than PRLIP1. The PRLIP1 family showed a tissue-specific expression pattern. Both PRLIP1 and PRLIP2 were specifically expressed in leaves and siliques, PRLIP1 additionally in stems and flowers. The expression of PRLIP6 and PRLIP4 was root specific, whereas mRNA of PRLIP5 and PRLIP7 were not detected in any of these tissues. The more distantly related genes PRLIP3, PRLIP9, and PRLIP8 were found on chromosomes 2, 4, and 5, respectively. The expression level of PRLIP3 was checked and found constitutive during the different stress conditions tested. The PRLIP1 gene was overexpressed in Escherichia coli, and the resulting PRLIP1 protein showed esterase activity on p-nitrophenyl-butyrate and allowed the growth of the bacteria on lipidic substrates such as Tween20 or Tween80. PMID- 12913178 TI - Expression of 1L-myoinositol-1-phosphate synthase in organelles. AB - We have studied the expression of 1L-myoinositol-1-phosphate synthase (MIPS; EC 5.5.1.4) in developing organs of Phaseolus vulgaris to define genetic controls that spatially regulate inositol phosphate biosynthesis. MIPS, the pivotal biosynthetic enzyme in inositol metabolism, is the only enzyme known to catalyze the conversion of glucose 6-phosphate to inositol phosphate. It is found in unicellular and multicellular eukaryotes and has been isolated as a soluble enzyme from both. Thus, it is widely accepted that inositol phosphate biosynthesis is largely restricted to the cytosol. Here, we report findings that suggest the enzyme is also expressed in membrane-bound organelles. Microscopic and biochemical analyses detected MIPS expression in plasma membranes, plastids, mitochondria, endoplasmic reticula, nuclei, and cell walls of bean. To address mechanisms by which the enzyme could be targeted to or through membranes, MIPS genes were analyzed for sorting signals within primary structures and upstream open reading frames that we discovered through our sequence analyses. Comprehensive computer analyses revealed putative transit peptides that are predicted to target the enzyme to different cellular compartments. Reverse transcriptase PCR experiments suggest that these putative targeting peptides are expressed in bean roots and leaves. PMID- 12913179 TI - Enhanced formaldehyde detoxification by overexpression of glutathione-dependent formaldehyde dehydrogenase from Arabidopsis. AB - The ADH2 gene codes for the Arabidopsis glutathione-dependent formaldehyde dehydrogenase (FALDH), an enzyme involved in formaldehyde metabolism in eukaryotes. In the present work, we have investigated the potential role of FALDH in detoxification of exogenous formaldehyde. We have generated a yeast (Saccharomyces cerevisiae) mutant strain (sfa1Delta) by in vivo deletion of the SFA1 gene that codes for the endogenous FALDH. Overexpression of Arabidopsis FALDH in this mutant confers high resistance to formaldehyde added exogenously, which demonstrates the functional conservation of the enzyme through evolution and supports its essential role in formaldehyde metabolism. To investigate the role of the enzyme in plants, we have generated Arabidopsis transgenic lines with modified levels of FALDH. Plants overexpressing the enzyme show a 25% increase in their efficiency to take up exogenous formaldehyde, whereas plants with reduced levels of FALDH (due to either a cosuppression phenotype or to the expression of an antisense construct) show a marked slower rate and reduced ability for formaldehyde detoxification as compared with the wild-type Arabidopsis. These results show that the capacity to take up and detoxify high concentrations of formaldehyde is proportionally related to the FALDH activity in the plant, revealing the essential role of this enzyme in formaldehyde detoxification. PMID- 12913181 TI - The Chlamydomonas reinhardtii cia3 mutant lacking a thylakoid lumen-localized carbonic anhydrase is limited by CO2 supply to rubisco and not photosystem II function in vivo. AB - The Chlamydomonas reinhardtii cia3 mutant has a phenotype indicating that it requires high-CO(2) levels for effective photosynthesis and growth. It was initially proposed that this mutant was defective in a carbonic anhydrase (CA) that was a key component of the photosynthetic CO(2)-concentrating mechanism (CCM). However, more recent identification of the genetic lesion as a defect in a lumenal CA associated with photosystem II (PSII) has raised questions about the role of this CA in either the CCM or PSII function. To resolve the role of this lumenal CA, we re-examined the physiology of the cia3 mutant. We confirmed and extended previous gas exchange analyses by using membrane-inlet mass spectrometry to monitor(16)O(2),(18)O(2), and CO(2) fluxes in vivo. The results demonstrate that PSII electron transport is not limited in the cia3 mutant at low inorganic carbon (Ci). We also measured metabolite pools sizes and showed that the RuBP pool does not fall to abnormally low levels at low Ci as might be expected by a photosynthetic electron transport or ATP generation limitation. Overall, the results demonstrate that under low Ci conditions, the mutant lacks the ability to supply Rubisco with adequate CO(2) for effective CO(2) fixation and is not limited directly by any aspect of PSII function. We conclude that the thylakoid CA is primarily required for the proper functioning of the CCM at low Ci by providing an ample supply of CO(2) for Rubisco. PMID- 12913180 TI - Aluminum-induced gene expression and protein localization of a cell wall associated receptor kinase in Arabidopsis. AB - Here, we report the aluminum (Al)-induced organ-specific expression of a WAK1 (cell wall-associated receptor kinase 1) gene and cell type-specific localization of WAK proteins in Arabidopsis. WAK1-specific reverse transcriptase-polymerase chain reaction analysis revealed an Al-induced WAK1 gene expression in roots. Short- and long-term analysis of gene expression in root fractions showed a typical "on" and "off" pattern with a first peak at 3 h of Al exposure followed by a sharp decline at 6 h and a complete disappearance after 9 h of Al exposure, suggesting the WAK1 is a further representative of Al-induced early genes. In shoots, upon root Al exposure, an increased but stable WAK1 expression was observed. Using confocal microscopy, we visualized Al-induced closure of leaf stomata, consistent with previous suggestions that the Al stress primarily experienced in roots associated with the transfer of root-shoot signals. Elevated levels of WAK protein in root cells were observed through western blots after 6 h of Al exposure, indicating a lag time between the Al-induced WAK transcription and translation. WAK proteins are localized abundantly to peripheries of cortex cells within the elongation zone of the root apex. In these root cells, disintegration of cortical microtubules was observed after Al treatment but not after the Al analog lanthanum treatments. Tip-growing control root hairs, stem stomata, and leaf stomatal pores are characterized with high amounts of WAKs, suggesting WAKs are accumulating at plasma membrane domains, which suffer from mechanical stress and lack dense arrays of supporting cortical microtubules. Further, transgenic plants overexpressing WAK1 showed an enhanced Al tolerance in terms of root growth when compared with the wild-type plants, making the WAK1 one of the important candidates for plant defense against Al toxicity. PMID- 12913183 TI - Nondopaminergic symptomatic therapies for Parkinson's disease: turn on or turn off? PMID- 12913184 TI - Unicorns, dragons, polymyositis, and other mythological beasts. PMID- 12913185 TI - Western Pacific ALS/PDC and flying foxes: What's next? PMID- 12913186 TI - Adenosine A(2A) receptor antagonist treatment of Parkinson's disease. AB - BACKGROUND: Observations in animal models suggest that A(2A) antagonists confer benefit by modulating dopaminergic effects on the striatal dysfunction associated with motor disability. This double-blind, placebo-controlled, proof-of-principle study evaluated the pathogenic contribution and therapeutic potential of adenosine A(2A) receptor-mediated mechanisms in Parkinson disease (PD) and levodopa-induced motor complications. METHODS: Fifteen patients with moderate to advanced PD consented to participate. All were randomized to either the selective A(2A) antagonist KW-6002 or matching placebo capsules in a 6-week dose-rising design (40 and 80 mg/day). Motor function was rated on the Unified PD Rating Scale. RESULTS: KW-6002 alone or in combination with a steady-state IV infusion of each patient's optimal levodopa dose had no effect on parkinsonian severity. At a low dose of levodopa, however, KW-6002 (80 mg) potentiated the antiparkinsonian response by 36% (p < 0.02), but with 45% less dyskinesia compared with that induced by optimal dose levodopa alone (p < 0.05). All cardinal parkinsonian signs improved, especially resting tremor. In addition, KW 6002 prolonged the efficacy half-time of levodopa by an average of 47 minutes (76%; p < 0.05). No medically important drug toxicity occurred. CONCLUSIONS: The results support the hypothesis that A(2A) receptor mechanisms contribute to symptom production in PD and that drugs able to selectively block these receptors may help palliate symptoms in levodopa-treated patients with this disorder. PMID- 12913187 TI - Randomized trial of the adenosine A(2A) receptor antagonist istradefylline in advanced PD. AB - OBJECTIVE: To evaluate the safety and efficacy of the adenosine A(2A) receptor antagonist istradefylline (KW-6002) in patients with levodopa-treated Parkinson's disease (PD) with both motor fluctuations and peak-dose dyskinesias. METHODS: This was a 12-week, double-blind, randomized, placebo-controlled, exploratory study in which PD subjects with both motor fluctuations and peak-dose dyskinesias were randomized to treatment with placebo (n = 29), istradefylline up to 20 mg/day (n = 26), or istradefylline up to 40 mg/day (n = 28). There was no prespecified primary outcome measure, and 19 outcome variables were analyzed. RESULTS: As assessed by home diaries, subjects assigned to istradefylline experienced a mean (+/- SE) reduction in the proportion of awake time spent in the "off" state of 7.1 +/- 2.0% compared with an increase of 2.2 +/- 2.7% in the placebo group (p = 0.008). There was a decrease in "off" time of 1.2 +/- 0.3 hours in the istradefylline group compared with an increase of 0.5 +/- 0.5 hour in the placebo group (p = 0.004). Dyskinesia severity was unchanged, but "on" time with dyskinesia increased in the istradefylline group compared with the placebo group (percent, p = 0.002; hours, p = 0.001). No differences were observed in change in Unified Parkinson's Disease Rating Scale scores or Clinical Global Impression of Change. Twenty-four percent of placebo-assigned subjects and 20% of istradefylline-assigned subjects withdrew from the study. Both dose regimens of istradefylline were generally well tolerated, and nausea was the most common adverse event. CONCLUSION: Istradefylline was generally well tolerated and reduced "off" time as assessed by home diaries. Severity of dyskinesia was unchanged, but "on" time with dyskinesia increased. PMID- 12913188 TI - Neuropathological examination suggests impaired brain iron acquisition in restless legs syndrome. AB - OBJECTIVE: To assess neuropathology in individuals with restless legs syndrome (RLS). METHODS: A standard neuropathologic evaluation was performed on seven brains from individuals who had been diagnosed with RLS. The substantia nigra was examined in greater detail for iron staining and with immunohistochemistry for tyrosine hydroxylase and proteins involved in iron management. Five age-matched individuals with no neurologic history served as controls. RESULTS: There were no histopathologic abnormalities unique to the RLS brains. Tyrosine hydroxylase staining in the major dopaminergic regions appeared normal in the RLS brains. Iron staining and H-ferritin staining was markedly decreased in the RLS substantia nigra. Although H-ferritin was minimally detected in the RLS brain, L ferritin staining was strong. However, the cells staining for L-ferritin in RLS brains were morphologically distinct from those in the control brains. Transferrin receptor staining on neuromelanin-containing cells was decreased in the RLS brains compared to normal, whereas transferrin staining in these cells was increased. CONCLUSIONS: RLS may not be rooted in pathologies associated with traditional neurodegenerative processes but may be a functional disorder resulting from impaired iron acquisition by the neuromelanin cells in RLS. The underlying mechanism may be a defect in regulation of the transferrin receptors. PMID- 12913189 TI - Increased ventral striatal monoaminergic innervation in Tourette syndrome. AB - BACKGROUND: Excessive striatal dopaminergic innervation is suggested to underlie Tourette syndrome (TS). Prior imaging and postmortem studies yield conflicting data. METHODS: The authors used PET with the type 2 vesicular monoamine transporter ligand [(11)C]dihydrotetrabenazine (DTBZ) to quantify striatal monoaminergic innervation in patients with TS (n = 19) and control subjects (n = 27). Compartmental modeling was used to determine blood to brain ligand transport (K(1)) and tissue to plasma distribution volume (a measure of ligand binding) during continuous infusion of DTBZ. TS data were compared with control data using predefined regions of interest and on a voxel by voxel basis. RESULTS: There were no significant differences in ligand binding or ligand transport between patients with TS and control subjects in the dorsal striatum. With voxel by voxel analysis, there was increased DTBZ binding in the right ventral striatum. CONCLUSIONS: Previously reported differences between patients with TS and control subjects in dorsal striatal dopamine terminal markers may reflect medication induced regulation of terminal marker expression or be the result of intrinsic differences in striatal dopaminergic synaptic function. Increased right ventral striatal DTBZ binding suggests that abnormal ventral striatal dopaminergic innervation may underlie tics. PMID- 12913190 TI - Polymyositis: an overdiagnosed entity. AB - BACKGROUND: According to widely used criteria (Bohan and Peter criteria, 1975), dermatomyositis (DM) is differentiated from polymyositis (PM) only by skin changes. More recent criteria also include histopathologic characteristics enabling the distinction between PM and DM and the differentiation of sporadic inclusion body myositis (s-IBM) from PM. The authors investigated the applicability of diagnostic features for diagnosing PM and DM. METHODS: The authors performed a retrospective follow-up study of 165 patients with 1) a previous diagnosis of myositis; 2) subacute onset of symmetric, proximal weakness; and 3) an evaluation between 1977 and 1998 excluding other neuromuscular disorders. RESULTS: The diagnoses at initial evaluation based on clinical, laboratory, and histopathologic criteria were PM, 9 (5%); DM, 59 (36%; 54 isolated, 3 with associated connective tissue disease [CTD], 2 with associated malignancy); unspecified myositis (perimysial/perivascular infiltrates, no PM or DM), 65 (39%; 38 isolated myositis, 26 with associated CTD, 1 with malignancy); and possible myositis (necrotizing myopathy, no inflammatory infiltrates), 32 (19%; 29 isolated myositis, 3 with associated CTD). At follow-up evaluation, five of the nine patients with PM had typical s-IBM features. None of the remaining four patients complied with the assumed typical signs of PM. Ten of the 38 patients with isolated unspecified myositis had been diagnosed with a CTD. CONCLUSIONS: Polymyositis is an overdiagnosed entity. At evaluation, more than half the patients with autoimmune myositis cannot be specifically diagnosed with polymyositis or dermatomyositis. A quarter of patients with isolated unspecified myositis subsequently developed connective tissue disease. PMID- 12913191 TI - MAP kinase phosphatase-1 is induced in abnormal fibers in inclusion body myositis. AB - OBJECTIVE: To investigate alterations in protein kinases and phosphatases that regulate the activity of mitogen activated protein kinase (MAPK) in sporadic inclusion body myositis (IBM). BACKGROUND: In vacuolated fibers in IBM, several studies reported upregulation of the extracellular regulated kinase (ERK) subclass of MAPK family. Whereas MAPK kinases (MKK) activate MAPK, MAPK phosphatases (MKP) inactivate MAPK. MKP-1 is involved in muscle fiber differentiation and it is downregulated during myotube formation. METHODS: Immunolocalization of MKK1 through MKK4 and MKP-1 to MKP-3 was tested in muscle specimens from 10 patients with IBM and controls. RESULTS: In IBM, strong and focal deposits of MKP-1 were observed in vacuolated fibers. The MKP-1-positive deposits were colocalized with ERK. MKP-2, MKP-3, and MKK were not associated with vacuolated fibers. CONCLUSIONS: In IBM, MKP-1 is abnormally induced in vacuolated fibers probably to inactivate ERK. Although direct activators other than those tested in the current study might induce ERK, the absence of activation of MKK suggests that the aggregation of ERK protein itself causes the seeming upregulation of the protein kinase in IBM. Like ERK and its nuclear substrate, MKP-1 is an enzyme that forms aggregates in vacuolated fibers and is involved in myogenesis. PMID- 12913192 TI - Mobius syndrome redefined: a syndrome of rhombencephalic maldevelopment. AB - OBJECTIVE: To investigate the variable clinical picture of Mobius syndrome (MIM no. 157900) and to further understand the pathogenesis of the disorder. METHODS: A standardized questionnaire was submitted to 37 Dutch patients with Mobius syndrome. All underwent standardized neurologic examination with special attention to cranial nerve functions, motor skills, and facial and limb anomalies. RESULTS: Of 37 patients with facial paresis, 97% had bilateral and 3% had unilateral ocular abduction weakness. Further analysis showed isolated abducens nerve palsy in 9%, a conjugated horizontal gaze paresis in 48%, features of Duane retraction syndrome in 34%, and congenital fibrosis of the extraocular muscles in 9%. Other signs included lingual involvement (77%), dysfunction of palate and pharynx (56%), general motor disability (88%), poor coordination (83%), and respiratory abnormalities (19%). CONCLUSION: Mobius syndrome is more than a cranial nerve or nuclear developmental disorder. It is a syndrome of rhombencephalic maldevelopment involving predominantly motor nuclei and axons, as well as traversing long tracts. The authors also noted gaze palsies, Duane retraction syndrome, feeding and respiratory problems, and poor motor development, suggesting a regional developmental disorder. PMID- 12913193 TI - Central Horner's syndrome with contralateral ataxic hemiparesis: a diencephalic alternate syndrome. AB - OBJECTIVE: To assess whether thalamic strokes presenting with a central Horner's syndrome (HS) show specific clinicoanatomic patterns. METHODS: From the Lausanne Stroke Registry (period 1993 to spring 2002), the authors selected all patients with thalamic stroke presenting with ipsilateral HS. Patients with complete infarction of the posterior cerebral artery territory, with involvement of middle cerebral artery territory or bilateral lesions, were excluded. Lesions on brain MRI were correlated with standard neuroanatomic templates. RESULTS: Nine patients with thalamic infarction presenting with central HS were found; all showed contralateral ataxic hemiparesis (AH). Lesions involved the anterior or paramedian thalamus and extended to the hypothalamic or rostral paramedian mesencephalic area in all but one subject. Associated clinical signs included dysphasia (two patients), somnolence (six), vertical gaze paresis (two), asterixis (two), and hemihypesthesia (three). CONCLUSION: The alternate clinical pattern of central HS with contralateral AH is a stroke syndrome of the diencephalic-mesencephalic junction, resulting from the involvement of the common arterial supply to the paramedian/anterior thalamus, the posterior hypothalamus and the rostral paramedian midbrain. PMID- 12913194 TI - Amelioration of osteoporosis and hypovitaminosis D by sunlight exposure in stroke patients. AB - BACKGROUND: The authors' previous investigations have disclosed low serum 25 hydroxyvitamin D (25-OHD) concentrations in 45 patients during long-term hospitalization following stroke (mean 5.9 ng/mL). This 25-OHD deficiency resulted from sunlight deprivation. OBJECTIVE: To evaluate the efficacy of sunlight exposure in increasing serum 25-OHD, in reducing the severity of osteoporosis in bone mineral density (BMD), and in decreasing the risk of hip fractures in chronically hospitalized, disabled stroke patients. METHODS: In a 12 month randomized and prospective study of stroke patients, 129 received regular sunlight exposure for 12 months, and the remaining 129 (sunlight-deprived) did not. RESULTS: At baseline, patients of both groups showed vitamin D deficiency. BMD increased by 3.1% in the sunlight-exposed group and decreased by 3.3% in the sunlight-deprived group (p = 0.0001). 25-OHD level increased by fourfold in the sunlight-exposed group. Six patients sustained hip fractures on the hemiplegic side in the sunlight-deprived group, and one hip fracture occurred among the sunlight-exposed group (p = 0421; odds ratio = 6.1). CONCLUSION: Sunlight exposure can increase the BMD of vitamin D-deficient bone by increasing 25-OHD concentration. PMID- 12913195 TI - Prediction of poststroke dementia. AB - OBJECTIVE: To investigate prospectively the frequency and clinical determinants of poststroke dementia (PSD) in a cohort of consecutive ischemic stroke inpatients in southern Taiwan. METHODS: A standard stroke evaluation protocol was conducted at admission and 3 months after an ischemic stroke. The protocol included clinical, neurologic, neurobehavioral, and functional assessments as well as neuroimaging examinations. Diagnoses were made according to the Neurologic Adaptation of the 10th edition of the International Classification of Diseases criteria for dementia. RESULTS: Excluding patients with prestroke dementia, a total of 283 patients were surveyed at 3 months after stroke; 26 (9.2%) of them met the criteria for PSD. The correlates of PSD in logistic regression analyses were age 65 years or older (odds ratio [OR] 6.6) vs <65 years, previous occupation as a laborer (OR 3.3), prior stroke (OR 3.1), left carotid vascular territory (OR 12.5) vs vertebrobasilar and unknown territories, moderate to severe stroke severity (OR 3.4), and cognitive impairment (OR 4.5) and poorer functional status at admission (OR 4.5). Based on the significant predictors identified, the logistic regression model correctly classified PSD in 93.4% of subjects. CONCLUSION: The lower frequency of PSD in this study from Taiwan compared with previous studies from Western countries may have been due to the relatively younger age of the elderly population and the use of stricter diagnostic criteria. PMID- 12913196 TI - Survival in frontotemporal dementia. AB - OBJECTIVES: To establish survival in patients with pathologically confirmed frontotemporal dementia (FTD) and to determine whether clinical or pathologic subtype affects prognosis. METHODS: The authors reviewed the presenting clinical features of 61 patients with dementia and pathologically confirmed FTD studied in Sydney (n = 31) and Cambridge (n = 30) over a 10-year period. Data were available on time of symptom onset, diagnosis, institutionalization, and death. Cases were classified pathologically as tau-positive and tau-negative. RESULTS: Of the 61 patients with FTD, 26 presented with frontal variant (fvFTD), 9 with semantic dementia, 8 with progressive nonfluent aphasia (PNFA), 9 with associated motor neuron disease (FTD-MND), and 9 with corticobasal degeneration features. There was no difference between the groups in age at symptom onset (overall mean 58.5 +/- 7.8 years), but at diagnosis the PNFA (68.3 +/- 2.7) group was significantly older than the fvFTD (59.9 +/- 7.4) and FTD-MND (57.7 +/- 7.9) groups. The median survival from symptom onset and from diagnosis was 6 +/- 1.1 years (95% CI) for fvFTD and 3 +/- 0.4 years for FTD-MND. Survival across subgroups was equivalent except for the FTD-MND group, which had significantly shorter survival. Cases with tau-positive pathology had an older age at onset and a significantly better prognosis: median survival 9.0 +/- 0.9 years vs 5.0 +/- 1.1 years. CONCLUSIONS: FTD is a malignant disorder with limited life expectancy. FTD-MND has the shortest duration both before and after diagnosis. Tau-positivity is associated with a more slowly progressive form of FTD. PMID- 12913197 TI - 14-3-3 protein in the CSF of patients with rapidly progressive dementia. AB - BACKGROUND: The presence of 14-3-3 protein in the CSF has been described to have high sensitivity and specificity for Creutzfeldt-Jakob disease (CJD). OBJECTIVE: To relate 14-3-3 protein in the CSF with the clinical diagnoses of diseases causing rapidly progressive dementia. METHODS: The authors studied 46 patients with rapidly progressive dementia that was classified into three diagnostic groups: definitive or probable CJD, possible CJD, and other diagnoses. The definitive or probable CJD group comprised 17 patients (3 definitive sporadic, 1 probable iatrogenic, 3 familial, and 10 probable sporadic CJD cases), the possible CJD group was composed of 7 patients, and the group with other diagnoses had 22 patients. Detection of the 14-3-3 protein was done by the immunoblotting method. RESULTS: In the definitive or probable CJD group, the test for 14-3-3 protein in CSF was positive in 14 (82%) cases, whereas 3 patients (1 probable sporadic and 2 familial cases) had negative results. CSF was positive for 14-3-3 protein in three of seven cases with possible CJD (42%). In the group with other diagnoses, three individuals had false-positive results (13%). Their diagnoses were definitive Alzheimer's disease, hypercalcemia, and multiple intracerebral hemorrhages. CONCLUSIONS: The detection of 14-3-3 protein in CSF is a useful in vivo diagnostic test for CJD and, when used in the appropriate clinical context, shows a good correlation to CJD. The presence of the 14-3-3 protein in the CSF reinforces the CJD clinical diagnosis but may not be able to differentiate CJD from other causes of rapidly progressive dementia in everyday clinical practice. PMID- 12913198 TI - Different patterns of N-acetylaspartate loss in subcortical ischemic vascular dementia and AD. AB - OBJECTIVES: 1) To determine the regional pattern of reduced N-acetylaspartate (NAA) in subcortical ischemic vascular dementia (SIVD); 2) to explore the relationship between NAA reduction and subcortical vascular disease; and 3) to test if MR spectroscopic imaging (MRSI) in combination with structural MRI improves differentiation between SIVD and Alzheimer disease (AD). METHODS: Thirteen patients with SIVD (71 +/- 8 years old) and 43 patients with AD of comparable age and dementia severity were studied using MRSI and MRI. Patients were compared to 52 cognitively normal subjects with and without lacunes. RESULTS: Compared to controls, patients with SIVD had lower NAA by 18% (p < 0.001) in frontal cortex and by 27% (p < 0.003) in parietal cortex, but no significant NAA reduction in white matter and medial temporal lobe. Compared to patients with AD, patients with SIVD had lower NAA by 13% (p < 0.02) in frontal cortex and by 20% (p < 0.002) in left parietal cortex. Cortical NAA decreased in SIVD with increasing white matter lesions (r = 0.54, p < 0.02) and number of lacunes (r = 0.59, p < 0.02). Thalamic lacunes were associated with greater NAA reduction in frontal cortex than were lacunes outside the thalamus (p < 0.02) across groups, after adjusting for cognitive impairments. Adding parietal NAA to MRI-derived hippocampal atrophy improved separation between SIVD and AD (p = 0.02) from 79 to 89%. CONCLUSIONS: These results emphasize the importance of cortical dysfunction as a factor in SIVD and indicate a characteristic pattern of metabolite change that might serve as a basis for improved diagnosis. PMID- 12913199 TI - Dysplasia: a common finding in intractable pediatric temporal lobe epilepsy. AB - BACKGROUND: Risk factors for temporal lobe epilepsy (TLE) include history of CNS infection, family history of epilepsy, and history of febrile convulsions (FC). Pre-existing cortical dysplasia (CD) may also predispose to refractory TLE, independent of other risk factors for epilepsy. METHODS: The authors reviewed the neuropathologic features of surgical tissue from temporal lobectomies of 33 pediatric patients with refractory TLE, with and without a history of epilepsy risk factors. RESULTS: CD was found in 64% (21/33) of all patients with refractory TLE, including 73% (11/15) patients with a history of FC, 66% (2/3) patients with CNS infections, and 83% (5/6) patients with a family history of epilepsy. Disrupted cortical lamination, dystrophic and maloriented neurons, and balloon cells characterized the CD found in the temporal neocortex. CONCLUSION: CD was seen in 21 of 33 surgical specimens from children with refractory TLE, including those with and without other epilepsy risk factors. PMID- 12913200 TI - Analysis of MRI patterns aids prediction of progression in X-linked adrenoleukodystrophy. AB - BACKGROUND: X-linked adrenoleukodystrophy (X-ALD) has variants with widely different outcomes, hampering clinical counseling and evaluation of therapies. OBJECTIVE: To evaluate the degree to which MRI patterns can predict lesion progression. METHODS: Two hundred six boys and men with cerebral X-ALD (median age 12.2 years, mean age 18.5 years, age range 1.7 to 73.8 years) were studied. In 140 individuals, follow-up MRI were available. Data after bone marrow transplantation (BMT) were excluded. The patterns of MRI abnormalities were subdivided into five groups based on the anatomic location of the initial T2 signal hyperintensity (pattern 1: parieto-occipital white matter, pattern 2: frontal white matter, pattern 3: corticospinal tract, pattern 4: cerebellar white matter, pattern 5: concomitant parieto-occipital and frontal white matter). The X ALD MRI Severity Scale, a 34-point scale previously described, was used in the analysis. RESULTS: Pattern 1 patients had rapid progression if contrast enhancement was present and if the MRI abnormality manifested at an early age. The latter was also true for pattern 2 patients. Based on these variables, predictive formulas were constructed for these two patterns using multiple regressions. MRI progression was much slower in pattern 3 and 4 patients, whereas in the few pattern 5 patients, it was more rapid than in any other of the patterns. Patterns 1 and 5 occurred mainly in childhood, patterns 2 and 4 in adolescence, and pattern 3 in adults. CONCLUSIONS: MRI progression in X-ALD depends on patient age, initial MRI Severity Scale score, and anatomic location of the lesion. When used in combination, these data aid the prediction of disease course and the selection of patients for BMT. PMID- 12913202 TI - Rescue of deep coma from sinus thrombosis. PMID- 12913201 TI - A self-administered screener for migraine in primary care: The ID Migraine validation study. AB - BACKGROUND: Migraine is a highly prevalent and disabling illness that remains substantially undiagnosed in primary care. Because of the potential value of a screening tool, the current study was designed to establish the validity and reliability of a brief, self-administered migraine screener in patients with headache complaints in the primary care setting. METHODS: A total of 563 patients presenting for routine primary care appointments and reporting headaches in the past 3 months completed a self-administered migraine screener. All patients were then referred for an independent diagnostic evaluation by a headache expert, of whom 451 (80%) completed a full evaluation. Migraine diagnosis was assigned based on International Headache Society criteria after completing a semi-structured diagnostic interview. RESULTS: Of nine diagnostic screening questions, a three item subset of disability, nausea, and sensitivity to light provided optimum performance, with a sensitivity of 0.81 (95% CI, 0.77 to 0.85), a specificity of 0.75 (95% CI, 0.64 to 0.84), and positive predictive value of 0.93 (95% CI, 89.9 to 95.8). Test-retest reliability was good, with a kappa of 0.68 (95% CI, 0.54 to 0.82). The sensitivity and specificity of the three-item migraine screener was similar regardless of sex, age, presence of other comorbid headaches, or previous diagnostic status. CONCLUSIONS: The three-item ID Migraine migraine screener was found to be a valid and reliable screening instrument for migraine headaches. Its ease of use and operating characteristics suggest that it could significantly improve migraine recognition in primary care. PMID- 12913203 TI - GNE mutations causing distal myopathy with rimmed vacuoles with inflammation. AB - The authors describe a family in which two individuals have clinical distal myopathy with rimmed vacuoles (DMRV). While the clinical and most of the pathologic features in these patients were compatible with a diagnosis of DMRV, the presence of inflammatory changes in the connective tissue between muscle fibers was not. Gene analysis revealed a compound heterozygous mutation in these individuals, characterized by V572L and I472T. PMID- 12913204 TI - Biomagnification of cycad neurotoxins in flying foxes: implications for ALS-PDC in Guam. AB - Beta-methylamino-L-alanine (BMAA) occurs in higher levels in museum specimens of the Guamanian flying fox than in the cycad seeds the flying foxes feed on, confirming the hypothesis that cycad neurotoxins are biomagnified within the Guam ecosystem. Consumption of a single flying fox may have resulted in an equivalent BMAA dose obtained from eating 174 to 1,014 kg of processed cycad flour. Traditional feasting on flying foxes may be related to the prevalence of neuropathologic disease in Guam. PMID- 12913205 TI - The potential value of ultrasonography in the evaluation of carpal tunnel syndrome. AB - The authors compared ultrasonography with electrophysiology for the diagnosis of carpal tunnel syndrome (CTS) on 110 clinically affected wrists. An increased cross sectional area in the proximal carpal tunnel larger than 0.11 cm(2) in combination with compression signs on longitudinal scans proved to be highly predictive for CTS (sensitivity, 89.1%; specificity, 98.0%). Ultrasound was comparable to electrophysiology in the diagnosis of CTS, and in 35% of cases changes in morphology suggested a specific therapeutic strategy. PMID- 12913206 TI - A neonatal form of glycogen storage disease type IV. AB - We report of an infant with neonatal glycogen storage disease type IV (GSD IV) who was examined for severe hypotonia and cardiomyopathy. On the muscle biopsy there were many fibers with diastase-resistant polyglucosan bodies. Glycogen branching enzyme (GBE1) activity in the muscle was markedly reduced. The infant had a homozygous single nucleotide deletion in the open reading frame of GBE1 gene. PMID- 12913208 TI - Prolonged treatment for acute symptomatic refractory status epilepticus: outcome in children. AB - High-dose suppressive therapy (HDST) is used to treat refractory status epilepticus (RSE). Prolonged therapy is required in some cases, and prognosis is important in making therapeutic decisions. The authors therefore studied the long term outcome in previously normal children who survived prolonged HDST for acute symptomatic RSE. All have intractable epilepsy, and none returned to baseline. PMID- 12913207 TI - Rasmussen encephalitis associated with Parry-Romberg syndrome. AB - Parry-Romberg syndrome is a rare disorder associated with unilateral facial atrophy involving skin, subcutaneous tissue, skeletal muscle, and bone. Occasionally, there is CNS involvement with epilepsy being the most common CNS manifestation. The authors report a child with Parry-Romberg syndrome with a course strongly suggestive of Rasmussen encephalitis. The boy underwent hemispherectomy, and pathology showed the typical findings of Rasmussen encephalitis, suggesting that these two conditions may share common etiologic factors. PMID- 12913209 TI - Video EEG monitoring prior to vagal nerve stimulator implantation. AB - Vagal nerve stimulation (VNS) is a safe alternative therapy for epilepsy but may have rare significant complications. There is no consensus regarding the necessity of video-EEG monitoring to characterize events before the VNS implantation. The authors discuss four patients who were inappropriately referred for or implanted with VNS without any previous video-EEG monitoring, in the context of their entire case experience. PMID- 12913210 TI - A novel autosomal dominant limb-girdle muscular dystrophy (LGMD 1F) maps to 7q32.1-32.2. AB - In 2001, the authors described the clinical features of a genetically distinct autosomal dominant limb-girdle muscular dystrophy (LGMD; LGMD 1F). Using a genome wide screen with more than 400 microsatellite markers, the authors identified a novel LGMD disease locus at chromosome 7q32.1-32.2. Within this chromosomal region, filamin C, a gene encoding actin binding protein highly expressed in muscle, was an obvious candidate gene; however, the authors did not detect any defects in filamin C or its protein product. PMID- 12913211 TI - Strong association of the Saitohin gene Q7 variant with progressive supranuclear palsy. AB - Recent reports are inconclusive in showing that the Q7R polymorphism of the novel Saitohin gene, nested in intron 9 of the tau gene, is associated with AD. The authors show that this polymorphism is in complete linkage disequilibrium with the extended tau H1/H2 haplotype and that the Q variant and QQ genotype of Q7R are strongly associated with progressive supranuclear palsy, implicating it as a possibly important pathogenic candidate. PMID- 12913212 TI - A family with seizures and minor features of tuberous sclerosis and a novel TSC2 mutation. AB - The authors studied nine members of a family that demonstrated a limited form of tuberous sclerosis complex (TSC). Cutaneous findings were limited to hypopigmented macules in four patients. Five family members had recurrent seizures, and three of these had migrational defects of the cerebral mantle. Mutational analysis of TSC2 indicated the presence of the novel missense change 3106T-->C, 1036S-->P in all family members with seizures. The findings suggest that this mild variant form of TSC is due to a novel TSC2 mutation. PMID- 12913213 TI - Cerebellar activation in opsoclonus: an fMRI study. AB - It is controversial whether opsoclonus is a cerebellar or brainstem disorder. Two patients whose opsoclonus largely disappeared on eye closure underwent fMRI. A comparison of these two states revealed neither vermal nor brainstem activation but rather a bilateral activation in the deep cerebellar nuclei in excess of what the authors found in healthy subjects. The results support a crucial role of the fastigial nucleus in opsoclonus. PMID- 12913214 TI - Eosinophilic fasciitis: MRI evaluation. PMID- 12913215 TI - The site of lesion in "vestibular neuritis": study by galvanic VEMP. PMID- 12913216 TI - HIV-1-related spontaneous episodic hypothermia. PMID- 12913217 TI - Levetiracetam as a treatment for tardive dyskinesia: a case report. PMID- 12913218 TI - Hemicraniectomy as a successful treatment of mass effect in acute disseminated encephalomyelitis. PMID- 12913219 TI - Trigeminal nerve stimulation for epilepsy. PMID- 12913220 TI - Pathological gambling associated with dopamine agonist therapy in Parkinson's disease. PMID- 12913221 TI - Migraine and autonomic nervous system function: a population-based, case-control study. PMID- 12913222 TI - The association of incident dementia with mortality in PD. PMID- 12913223 TI - Trochleitis and migraine headache. PMID- 12913224 TI - Dramatic response to chemotherapy in oligodendroglial gliomatosis cerebri. PMID- 12913225 TI - Self-aggregation inhibits the photonuclease activity of porphyrins. AB - A series of tentacle porphyrins having four aminoalkyl groups at the periphery was synthesized, and the DNA binding properties were investigated by absorption and circular dichroism (CD) spectroscopic methods. The aminopropyl chain was found to facilitate binding, and bisignate induced CD spectra revealed that the porphyrins are self-stacked on the DNA surface. The photonuclease activity of the tentacle porphyrins was also studied, and the aminopropylporphyrin showed the highest activity. The activity increased in proportion to the porphyrin load, but higher loads resulted in the decrease of activity. This inhibitory step corresponded to aggregation of the porphyrin. Thus, the aggregation was suggested to shield the inner porphyrin from the solvent, the production of active oxygen species being suppressed. PMID- 12913226 TI - Release from or through a wax matrix system. V. Applicability of the square-root time law equation for release from a wax matrix tablet. AB - To obtain basic and clear release properties, wax matrix tablets were prepared from a physical mixture of drug and wax powder at a fixed mixing ratio. Properties of release from the single flat-faced surface, curved side surface, and/or whole surface of the wax matrix tablet were examined. Then tortuosity and the applicability of Higuchi's square-root time law equation were examined. The Higuchi equation well analyzed the release processes of different release manners. However, the region fitted to the Higuchi equation differed with the release manner. Tortuosity obtained with release from the single flat-faced surface and curved side surface was comparable with that obtained with the release from a reservoir device tablet, whereas tortuosity obtained with release from the whole surface was larger. As the wax matrix tablets were prepared at a fixed mixing ratio, their internal structures should be similar. Therefore changes in the matrix volume or volume fraction with release were examined, and an extra volume where dissolved drug stray becomes large with release time in the case of release from the whole surface. These factors should be taken into account for evaluation of applicability and release properties. Furthermore, the entire release process should be analyzed using a combination of the square-root time law and other suitable equations in accordance with release manner or condition. PMID- 12913227 TI - Isolation of three marine prostanoids, possible biosynthetic intermediates for clavulones, from the Okinawan soft coral Clavularia viridis. AB - Three marine prostanoids, 1, 2, and 3, were isolated from the extract of the Okinawan soft coral Clavularia viridis. The structures of these compounds were assigned based on the results of spectroscopic analysis. Compound 1 was shown to be preclavulone-A methyl ester, and this is the first isolation of the ester of preclavulone-A as a natural product. Preclavulone-A is proposed to be the key intermediate in the biosynthesis of marine prostanoids exemplified by clavulones in C. viridis. The new prostanoid 3 was suggested to be a biosynthetic intermediate from preclavulone-A to clavulones, and a possible biogenetic pathway via 3 is proposed. PMID- 12913228 TI - Investigation and physicochemical characterization of vinpocetine-sulfobutyl ether beta-cyclodextrin binary and ternary complexes. AB - The purpose of this study was to investigate the interactions between vinpocetine (VP), sulfobutyl ether beta-cyclodextrin (SBEbetaCD) and the water-soluble polymers polyvinylpyrrolidone (PVP) and hydroxypropyl methylcellulose (HPMC). The water-soluble polymers were shown to improve the complexation efficiency of SBEbetaCD, and thus less SBEbetaCD was needed to prepare solid VP-SBEbetaCD complexes in the presence of the polymers. The interactions between VP and SBEbetaCD, with or without PVP or HPMC, were thoroughly investigated in aqueous solutions using the phase-solubility method as well as in the solid state. The amount of VP solubilized in water or aqueous polymer solution increased linearly with increasing SBEbetaCD concentration, demonstrating A(L)-type plots. We estimated the apparent stability constant (K(c)) at room temperature of VP SBEbetaCD binary complex to be 340 M(-1) and this value increased to 490 M(-1) or 390 M(-1), respectively, with the addition of PVP and HPMC, assuming a 1 : 1 VP SBEbetaCD molar ratio. Improvement in the K(c) values for ternary complexes clearly confirmed the benefit of the addition of water-soluble polymers to promote higher complexation efficiency. Solid VP-SBEbetaCD binary and ternary systems were prepared by physical mixing, kneading, coevaporation, and lyophilization methods and fully characterized by scanning electron microscopy, differential scanning calorimetry, and X-ray diffractometry. The results obtained suggest that coevaporation and lyophilization methods yield a higher degree of amorphous entities and indicated formation of VP-SBEbetaCD binary and ternary complexes. PMID- 12913229 TI - Dynamic flexibility of a peptide-binding groove of human HLA-DR1 class II MHC molecules: normal mode analysis of the antigen peptide-class II MHC complex. AB - Class II major histocompatibility complex (MHC) has tolerance for binding longer antigen peptides than those bound by class I MHC. In this paper, a normal mode analysis on HLA-DR1 class II MHC involving an antigen peptide indicated that the peptide-binding groove had some different dynamic characteristics from that of HLA-A2 class I MHC. The dynamic changes in the class I groove with removal of the bound peptide were limited primarily to the central region and the C-terminal side (corresponding to the C-terminal side of the bound peptide) of the groove, while the dynamic changes in the class II groove with removal of the bound peptide extended to the whole of the groove, and were especially remarkable around a strand located in the N-terminal side (corresponding to the N-terminal side of the bound peptide) of the groove. These results suggest that the N terminal side of the class II groove is more flexible than the same side of the class I groove, and this flexibility may allow some N-terminal residues of the bound peptide to extend outside the class II groove. Definite anti-correlative motions with removal of the bound peptide appeared between two alpha-helical regions of class II MHC as in the case of class I MHC. These motions of the class II groove may play an important role in obtaining "a flexible dynamic fit" against diverse longer peptides both of whose terminals extend outside the groove. PMID- 12913230 TI - Fentanyl and its analogue N-(1-phenylpyrazol-3-yl)-N-[1-(2-phenylethyl)-4 piperidyl]propanamide: 1H- and 13C-NMR spectroscopy, X-ray crystallography, and theoretical calculations. AB - The oxalate salts and free bases of fentanyl and N-[1-(2-phenylethyl)-4 piperidyl]-N-(1-phenyl-4-pyrazolyl)propanamide, a new lead compound for long acting analgesia, have been characterized by (1)H- and (13)C-NMR spectroscopy. The crystal structure of the hydrochloride of N-[1-(2-phenylethyl)-4-piperidyl]-N (1-phenyl-4-pyrazolyl)propanamide monohydrate has been determined. Two centrosymmetrically related cations, joined through C(phenyl)-H em leader pi contacts, encapsulate a large void that contains pairs of anions and bridged water molecules into a zero-dimensional (0D) supramolecular motif. The cations are linked to this framework via N(+)H em leader Cl(-) contacts. GIAO/B3LYP calculations have been carried out to compare the experimental (13)C chemical shifts with the absolute shieldings thus calculated. The protonation of both molecules takes place on the piperidine ring (axial protonation), as has been verified both in the solid state (X-ray) and in solution (NMR). PMID- 12913231 TI - Diterpenes from the roots of Euphorbia kansui and their in vitro effects on the cell division of Xenopus (part 2). AB - Four new ingenane-type diterpenes, 3-O-(2,3-dimethylbutanoyl)-13-O-dodecanoyl-20 O-acetylingenol (1), 3-O-(2,3-dimethylbutanoyl)-13-O-dodecanoyl-20-deoxyingenol (2), 3-O-(2E,4Z-decadienoyl)-20-deoxyingenol (3), and 3-O-(2E,4E-decadienoyl)-20 deoxyingenol (4), two new jatrophane-type diterpenes, kansuinins D (9) and E (10), and four known ingenane-type diterpenes were isolated from the root of Euphorbia kansui. Their structures were elucidated by spectroscopic and chemical analysis, and individual Xenopus cells at the blastular stage were cultured with the diterpenes to test for biological activity. 20-Deoxyingenol diterpenes 3 and 4 induced the greatest cell cleavage arrest (0.5 micro g/ml of each compound resulted in >75% cleavage arrest), but cell cleavage inhibitory activity became weak when C-16 had an acyl residue. In contrast, the jatrophane diterpene kansuinin D (9) showed no activity. PMID- 12913232 TI - Formulation study for lansoprazole fast-disintegrating tablet. I. Effect of compression on dissolution behavior. AB - Lansoprazole fast-disintegrating tablet (LFDT) is a new patient-friendly formulation of lansoprazole. Since lansoprazole is an antiulcer agent and is unstable under acidic conditions, we have developed LFDT as an orally disintegrating tablet containing enteric-coated microgranules. The effect of compression on dissolution behavior was investigated, as compression affected cleavage and crushing of the enteric layer. To decrease cleavage and crushing of the enteric layer, the effects of the combined ratio of methacrylic acid copolymer dispersion to ethyl acrylate-methyl methacrylate copolymer dispersion and the concentration of triethyl citrate on the dissolution in the acid stage and the dissolution in the buffer stage were evaluated. By adjusting the ratio of methacrylic acid copolymer dispersion to ethyl acrylate-methyl methacrylate copolymer dispersion to 9 : 1 and adding a 20% triethyl citrate concentration, sufficient flexibility of the enteric layer and sufficient stability against compression forces were achieved. Agglomeration of enteric-coated microgranules during the coating process was decreased at the optimized concentration of triethyl citrate and glyceryl monostearate. We compared the absorption properties of LFDT and lansoprazole capsules in dogs. The absorption profiles of LFDT were similar to those of lansoprazole capsules. PMID- 12913233 TI - Cytotoxic anthraquinones from the stems of Rubia wallichiana Decne. AB - From the stems of Rubia wallichiana DECNE, thirty-four structurally related compounds were isolated and identified. Three of them, namely rubiawallin-A (1), B (2), and -C (3), constitute the first report of their occurrence from the natural source. Their structures were determined by comprehensive analyses of their 1D and 2D NMR, and electron impact (EI) mass spectral data. Furthermore, an in vitro screening of cytotoxicity of the isolated compounds was also evaluated. Among the testing compounds, 1-hydroxy-2-hydroxymethyl-3-methoxyanthraquinone (4) demonstrated most effective cytotoxicity towards Hepa-3B and Colo-205 cells. PMID- 12913234 TI - The synthesis of isoquinoline alkaloid and its related compounds using alanine derivatives as chiral auxiliaries. AB - Chiral 1-substituted isoquinoline derivatives, which were obtained by the reaction using alanine derivatives as chiral auxiliaries, were transformed to (S) 2,3,9,10,11-pentamethoxyhomoprotoberberine (7) and a synthetic intermediate for O methylkreysigine (9) in good yields and high stereoselectivity. The corresponding chiral allyl derivative of isoquinoline was transformed to a pyrrolidinoisoquinoline (16) in a highly enantioselective manner. PMID- 12913235 TI - Four new cycloartane glycosides from Aquilegia vulgaris. AB - Four new cycloartane glycosides, named aquilegiosides G-J, were isolated from the dried aerial parts of Aquilegia vulgaris. Their structures were determined by spectroscopic analysis and chemical evidence. PMID- 12913236 TI - Triterpenes and triterpene saponins from the stems of Akebia trifoliata. AB - To characterize the stems of Akebia trifoliata chemically, a detailed phytochemical examination was carried out on A. trifoliata stems, with particular attention to the triterpene and triterpene saponin constituents, and resulted in the isolation of three new triterpenes (1-3) and three new triterpene saponins (11-13), together with seven known triterpenes (4-10) and 12 known triterpene saponins (14-25). The structures of the new compounds were determined on the basis of spectroscopic analysis, including two-dimensional NMR spectroscopic data, and the results of hydrolysis. Four saponins (22-25), which were obtained in good yields and were not isolated from Akebia quinata stems, are concluded to be applicable as marker compounds in chemically distinguishing between A. trifoliata and A. quinata by conventional TLC examination. To the best our knowledge, the current work is the first chemical investigation of A. trifoliata. PMID- 12913237 TI - Reaction of magnesium alkylidene carbenoids with lithium alpha-sulfonyl carbanions: a novel synthesis of tri- and tetra-substituted allenes from 1 chlorovinyl p-tolyl sulfoxides and sulfones. AB - Treatment of 1-chlorovinyl p-tolyl sulfoxides, which were synthesized from ketones and chloromethyl p-tolyl sulfoxide, with ethylmagnesium chloride or isopropylmagnesium chloride at below -78 degrees C gave magnesium alkylidene carbenoids in about 90% yields. The reaction of the generated carbenoids with lithium alpha-sulfonyl carbanions was found to afford tri- and tetra-substituted allenes. Both cyclic ketones and acyclic ketones were useful in this procedure. However, the 1-chlorovinyl p-tolyl sulfoxides derived from aldehydes gave only rearranged products, acetylenes, under the reaction conditions. The magnesium alkylidene carbenoid derived from an optically active 1-chlorovinyl p-tolyl sulfoxide was treated with lithium alpha-carbanion of 1-naphthyl phenyl sulfone; however, the obtained allene was found to be racemic. The mechanism of this reaction is also discussed. PMID- 12913238 TI - Synthesis and preliminary biological evaluation of a new pyridocarbazole derivative covalently linked to a thymidine nucleoside as a potential targeted antitumoral agent. I. AB - The therapy of human cancer is one of the more pursued goals by medicinal chemistry research. Most of the compounds clinically used as a treatment owe their efficacy to their cytotoxic interaction (direct or indirect) with nuclear DNA. This interaction results in the inhibition of DNA synthesis and the degradation of nucleic strands. Ellipticine is a naturally occurring 6H pyrido[4,3-b]carbazole alkaloid endowed with antitumor activity, and several ellipticine derivatives have been used in clinical trials. We previously reported some 1,4-dimethyl-9H-carbazole derivatives structurally related to ellipticine. The purpose of our research was to transform the pyridocarbazole in a prodrug so that it would have more penetration in the tumor cells and block their replication. Our prodrug is slowly hydrolyzed in human plasma in the corresponding acid. From these preliminary results, we deduce that our compound can block cellular replication. Our hypothesis is that the antitumoral activity is probably related to the induction of damage to DNA, without cellular lysis in the short term. PMID- 12913239 TI - Synthesis of 2-(pyrimidin-4-yl)indoles. AB - The synthesis of 2-substituted isomers of the meridianins, a familiy of bioactive indole alkaloids isolated from the tunicate Aplidium meridianum, was undertaken. The synthetic route comprises six steps, with a microwave promoted Fischer cyclization as the key reaction. PMID- 12913240 TI - Hydroxypropyl methylcellulose based cephalexin extended release tablets: influence of tablet formulation, hardness and storage on in vitro release kinetics. AB - The object of this study was to develop hydroxypropyl methylcellulose (HPMC) based cephalexin extended release tablet, which can release the drug for six hours in predetermined rate. Twenty-one batches of cephalexin tablets were prepared by changing various physical and chemical parameters, in order to get required theoretical release profile. The influences of HPMC, microcrystalline cellulose powder (MCCP), granulation technique, wetting agent and tablet hardness on cephalexin release from HPMC based extended release tablets were studied. The formulated tablets were also characterized by physical and chemical parameters. The dissolution results showed that a higher amount of HPMC in tablet composition resulted in reduced drug release. Addition of MCCP resulted in faster drug release. Tablets prepared by dry granulation was released the drug slowly than the same prepared with a wet granulation technique. Addition of wetting agent in the tablets prepared with dry granulation technique showed slower release. An increase in tablet hardness resulted in faster drug release. Tablets prepared with a wet granulation technique and having a composition of 9.3% w/w HPMC with a hardness of 10-12 kg/cm(2) gave predicted release for 6 h. The in vitro release data was well fit in to Higuchi and Korsmeyer-Peppas model. Physical and chemical parameters of all formulated tablets were within acceptable limits. One batch among formulated twenty-one batches was successful and showed required theoretical release. The effect of storage on in vitro release and physicochemical parameters of successful batch was studied and was found to be in acceptable limits. PMID- 12913242 TI - Cadinane-type sesquiterpenes from the roots of Taiwania cryptomerioides Hayata. AB - Five new cadinane-type sesquiterpenes, 15-acetoxy-T-muurolol (1), isokhusinodiol (2), cadin-10(14)-ene-4beta,5alpha-diol (3), cadinane-4beta,5alpha,10beta-triol (4), and muurolane-4beta,5beta,10beta-triol (5), together with five known compounds, T-cadinol (6), T-muurolol (7), alpha-cadinol (8), delta-cadinol (9), and khusinodiol (10), were isolated from the roots of Taiwania cryptomerioides. The structure of the new constituents were elucidated through chemical and spectral studies. PMID- 12913241 TI - Cyclodextrin-enclosed substances of Brazilian propolis. AB - By using beta-cyclodextrin-inclusion as a unique technique, an efficient separation of pharmacologically active phenolic compounds from Brazilian propolis was achieved to provide one new compound, 3-(3-hydroxy-3-methyl-butyl)-5-prenyl-4 hydroxycinnamic acid, together with two common cinnamic acid derivatives, artepillin C and capillartemisin A, and two known flavanols, aromadendrin and 3,5,7-trihydroxy-4'-methoxyflavanol. PMID- 12913243 TI - Two new bromotyrosine-derived metabolites from the sponge Psammaplysilla purpurea. AB - Two new bromotyrosine-derived metabolites (1, 2) have been isolated along with the known compounds 3,5-dibromo-4-methoxyphenylacetonitrile, 3-bromo-4 methoxyphenylacetonitrile, 3-bromo-4-hydroxyphenylacetonitrile, 1-hydroxyuracil, 1-methoxyhemibastadin 2, purpuramine H and a steroid 5alpha,8alpha epidioxycholest-6-en-3beta-ol from the sponge Psammaplysilla purpurea. Compounds 1 and 2 were characterized by interpretation of their spectral data. The antibacterial activity of these compounds is summarized. PMID- 12913244 TI - Synthesis and root growth-inhibitory activity of 2- and 3-(haloacetylamino)-3-(2 furyl)propanoic acids. AB - A convenient synthesis of 2- and 3-(chloroacetylamino)-3-(2-furyl)propanoic acids (6a, 7a) and their fluoro analogs were developed. Both 6a and 7a showed 51-55% root growth-inhibitory activity towards rape seedlings at the concentration of 1.0x10(-4) M. PMID- 12913245 TI - Norditerpenoid alkaloids from the processed tubers of Aconitum carmichaeli. AB - Four new and five known norditerpenoid alkaloids were isolated from the processed tubers of Aconitum carmichaeli. The new alkaloids are 14-O-cinnamoylneoline (3), 14-O-anisoylneoline (4) 14-O-veratroylneoline (5), and lipo-14-O anisoylbikhaconine (8). The known alkaloids are neoline (1), 14-O-acetylneoline (2), foresaconitine (6), crassicauline A (7), and lipohypaconitine (9). Alkaloids 2, 6, and 7 were isolated from this plant for the first time. The structures of the new alkaloids were established by spectroscopic and chemical methods. PMID- 12913246 TI - A new norsesquiterpenoid from Russula delica Fr. AB - Russulanorol (1), a new norsesquiterpenoid with a novel carbon skeleton, was isolated from the fruiting bodies of Russula delica FR. (Russulaceae) together with three known sesquiterpenoids. The structure of 1 was elucidated on the basis of spectral data and chemical transformation. Compound 1 is an equilibrium mixture of two stereoisomers (1a, b) on the C-11 acetal carbon. PMID- 12913247 TI - Insulinomimetic zinc(II) complexes with natural products: in vitro evaluation and blood glucose lowering effect in KK-Ay mice with type 2 diabetes mellitus. AB - In vitro insulinomimetic activities of Zn(II) complexes with three natural products, betaine, L-lactic acid, and D-(-)-quinic acid (qui), were found in rat adipocytes treated with epinephrine in terms of the inhibition of free fatty acid release. Based on the results, the blood glucose lowering effect in KK-A(y) mice with type 2 diabetes mellitus was observed by daily i.p. injections of a monomeric zinc(II) complex, Zn(qui)(2), for 13 d. PMID- 12913249 TI - A new phlorotannin from the brown alga Ecklonia stolonifera. AB - A new phlorotannin, named eckstolonol (1), was isolated from the EtOAc soluble fraction of the methanolic extract of the brown alga, Ecklonia stolonifera OKAMURA, along with three known phlorotannins, eckol (2), phlorofucofuroeckol A (3), and dieckol (4). The structure of eckstolonol was identified as 5,8,13,14 tetraoxa-pentaphene-1,3,6,9,11-pentaol on the basis of spectroscopic evidence. The new compound was found to be a radical scavenger on the 1,1-diphenyl-2 picrylhydrazyl (DPPH) radical. PMID- 12913248 TI - Polyhydroxylated steroids and other constituents of the soft coral Nephthea chabroli. AB - Polyhydroxylated steroids and other metabolites were isolated from the soft coral Nephthea chabroli collected in South China Sea. The structures of two new compounds were determined to be 24-methylcholesta-9(11), 24(28)-diene 3beta,12alpha,19-triol and 4alpha-methyl-3beta,14beta-dihydroxy-5alpha-ergost 24(28)-en-23-one on the basis of spectroscopic analysis. Cytotoxic activities of the polyhydroxylated sterols were evaluated on prostate carcinoma LNCaP cell line. PMID- 12913250 TI - Synthetic study of optically active 3-azabicyclo[3.3.0]octane-2,6,8-tricarboxylic acid. AB - Synthesis of (1R,2S,5S,6R,8S)-3-azabicyclo[3.3.0]octane-2,6,8-tricarboxylic acid (2) from trans-4-hydroxy-L-proline (5) was attempted. A Diels-Alder reaction of 3,4-dehydroproline derivative 9 and cyclopentadiene afforded a single stereoisomer 11. The Diels-Alder adduct was smoothly converted to the hydrochloride of 2 (24) via RuO(4) oxidation. Although some racemization of the material or product was observed during the synthetic processes, the amino acid 24 proved to be optically pure. PMID- 12913251 TI - Virus concentration using sulfonated magnetic beads to improve sensitivity in nucleic acid amplification tests. AB - To enhance the sensitivity of virus detection by polymerase chain reaction (PCR) and reverse-transcriptional (RT)-PCR, we developed a novel virus-concentration method using sulfonated (SO-) magnetic beads in the presence of divalent cations. In the presence of either Zn(2+) or Cu(2+) ions, we showed that SO-magnetic beads were able to concentrate non-enveloped model viruses, such as porcine parvovirus (PPV) and poliovirus, which were not concentrated by polyethyleneimine (PEI) magnetic beads.(1)) Using the SO-magnetic beads, the sensitivity of virus genome detection by PCR or RT-PCR can be enhanced. Therefore, an efficient virus concentration method using either SO-magnetic beads or PEI-magnetic beads enhances the sensitivity of virus detection by PCR or RT-PCR. PMID- 12913252 TI - Does peroxynitrite involve in the elevation of cellular glutathione induced by sodium nitroprusside (SNP) in RAW 264.7 cells? AB - The mechanism underlying the elevation of intracellular glutathione (GSH) in RAW 264.7 cells exposed to low-level sodium nitroprusside (SNP) was investigated by measuring the expression of mRNA for gamma-glutamylcysteine synthetase (gamma GCS), the rate-limiting enzyme of de novo GSH synthesis, and the GSH content. A significant elevation of expression of mRNA for gamma-GCS was observed at 3 h after exposure of the cells to SNP at a concentration of 0.25 mM. 2-(4 Carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (c-PTIO), N acetylcysteine (NAC), or ebselen (Ebs) significantly suppressed the elevations induced by SNP, suggesting that hydrogen peroxide or peroxynitrite (ONOO(-)) is involved in this event as a triggering molecule. Hydrogen peroxide itself, however, did not induce the elevation of gamma-GCS mRNA and glutathione. Chemiluminescenses induced by SIN-1, a chemical ONOO(-) donor, and ONOO(-) itself were completely blocked by Ebs. SIN-1 also significantly elevated the cellular glutathione level, and the elevation was absolutely blocked by Ebs. These results suggest that the elevation of intracellular GSH in RAW 264.7 cells exposed to low level SNP occurs via the de novo GSH pathway through transcriptional up regulation of the gamma-GCS gene induced by peroxynitrite molecule. PMID- 12913253 TI - Interleukin-10-induced CCR5 expression in macrophage like HL-60 cells: involvement of Erk1/2 and STAT-3. AB - As an immunosuppressive and anti-inflammatory cytokine, IL-10 was recently reported to play roles in CCR5 expression in human monocytes. CCR5 promoter regions contain Oct-2, TCF-1alpha, GATA, and STAT binding sites. Here, we studied the signals involved in the CCR5 expression in IL-10-stimulated cells using the HL-60 cell line. HL-60 cells were stimulated with PMA and differentiated to macrophage-like cells, then stimulated with IL-10. IL-10 induced significant expression of CCR5 protein and CCR5 mRNA in these cells. The induction of CCR5 by IL-10 was inhibited by a MEK-1 inhibitor, PD98059. In addition, IL-10 induced tyrosine (Tyr) phosphorylation of Erk, as well as serine (Ser) and Tyr phosphorylation of STAT-3. Tyr phosphorylation of Erk and Ser phosphorylation of STAT-3 were inhibited by PD98059, while Tyr phosphorylation of STAT-3 was not inhibited by PD98059. DNA binding activity of STAT-3 was observed by the stimulation with IL-10, which was inhibited by PD98059. These results first indicate that Erk1/2 and STAT-3 regulate CCR5 expression, and that Erk-mediated phosphorylation of Ser is required for full stimulation of STAT-3 in CCR5 expression. PMID- 12913254 TI - Change in the protein level of mevalonate pyrophosphate decarboxylase in tissues of mouse by pravastatin. AB - We previously reported that treatment of rats with a diet containing 0.1% pravastatin and 5% cholestyramine markedly increased mevalonate pyrophosphate decarboxylase (MPD) activity in liver crude extracts compared with nontreated rats. In this study, we examined the change in the protein level of MPD in the tissues of mice administered pravastatin. When MPD content in the tissues of nontreated mice was analyzed by quantitative immunoblotting, a single protein band with an apparent molecular weight of 46 kDa was detected in all tissues and the specific protein content of MPD in liver and kidney was markedly higher than that in other tissues. When MPD content in the tissues of pravastatin-treated mice was analyzed by immunoblotting, MPD was markedly increased (9-fold) only in the liver compared with nontreated mice. Next, when MPD activity was measured in the liver between nontreated and pravastatin-treated mice, MPD activity as well as protein levels were markedly increased (11-fold) in the liver of pravastatin treated mice compared with nontreated mice. These data suggest that a marked induction of MPD in the liver by pravastatin is responsible for the tissue specific effect of pravastatin. PMID- 12913255 TI - Analysis of angiotensin II mediated COX-2 downregulation in angiotensin II- or aldosterone-infused hypertensive rat. AB - The renin-angiotensin cascade plays an important role in blood pressure control and sodium homeostasis. This study investigated whether cyclooxygenase-2 expression is regulated in the kidney, in an angiotensin II- and aldosterone induced hypertension model. For this purpose, we treated male Sprague-Dawley rats (n=8 per group) with angiotensin II (9 mg/h, subcutaneously) for 14 d and aldosterone (0.75 mg/h, subcutaneously) for 42 d. Systolic blood pressure was significantly increased by angiotensin II (p<0.001) and by aldosterone (p<0.001). We found that angiotensin II downregulated cyclooxygenase-2 protein in the kidney cortex, whereas aldosterone showed no effect. These results indicate that angiotensin II may act directly to inhibit kidney cortex cyclooxygenase-2 protein expression, rather than acting via stimulation of aldosterone. PMID- 12913256 TI - Alleviating effects of si-ni-san, a traditional Chinese prescription, on experimental liver injury and its mechanisms. AB - The present study aims at examining the effects of Si-Ni-San, a prescription usually used for treating hepatitis in Traditional Chinese Medicine (TCM), on various experimental liver injury models and its mechanisms. The prescription showed significant hepatoprotection against CCl(4)-induced hepatic damage, both in vivo and in vitro. To the liver injury induced by Bacillus Calmette-Guerin (BCG) with lipopolysaccharide (LPS), Si-Ni-San also provided significant alleviation through enhancing nitric oxide (NO) release by macrophages. Against the liver injury induced by a delayed-type hypersensitivity reaction to picryl chloride (PCl-DTH), Si-Ni-San alleviated it remarkably when administered during either the induction or effector phase. A significant reduction of in-vitro hepatotoxicity, as measured by the inhibition of serum transaminase evaluation, was observed in nonparenchymal cells from liver-injured mice treated with Si-Ni San. Si-Ni-San facilitated apoptosis in nonparenchymal cells from liver-injured mice, as well as in spleen cells activated by PCl in vivo or by Con A in vitro. These results suggest that Si-Ni-San provides alleviating effects against liver injury through multiple mechanisms, including protection of the hepatocyte membrane, enhancement of NO release, and dysfunction of liver-infiltrating cells mainly through causing their apoptosis. PMID- 12913257 TI - Cardiovascular protective effects of traditional Chinese medicine bak foong pills in spontaneously hypertensive rats. AB - The cardiovascular protective effects of the traditional Chinese medicine Bak Foong Pills (BFP) were investigated. Spontaneously hypertensive rats (SHR) were treated (3 g/kg) over a 5-month period and blood pressure measurements periodically tested with a plethysmographic tail cuff. Following treatment, blood samples were analysed for serum electrolyte levels and lipid levels and brain tissue subjected to micro-array analysis. In vitro experiments were also conducted to identify possible direct vasorelaxatory effect. The results showed that BFP was able to significantly reduce both systolic and diastolic blood pressure by about 30 mmHg in SHR following 5 months of treatment, when compared to untreated animals. Investigation for possible mechanisms of actions revealed that BFP treated rats had elevated blood serum K(+) levels, and also demonstrated decreased serum triglyceride levels. Micro-array analysis of brain tissue showed altered expression of acetylcholine and lysosphingolipid receptor genes that are known to regulate blood pressure. In vitro experiments also showed that BFP caused a concentration-dependant vasorelaxation of isolated rat aortae when contracted with phenylepherine, which was partially inhibited by nitric oxide synthase inhibitor L-NAME (100 microM). These data suggest that BFP is able to significantly reduce hypertension in SHR through mechanisms probably involving a combination of increased serum K(+), vasorelaxatory action, reduced serum triglyceride and altered gene regulation in the higher centres. PMID- 12913258 TI - Antidiabetic effects of chitosan oligosaccharides in neonatal streptozotocin induced noninsulin-dependent diabetes mellitus in rats. AB - The antidiabetic effect of chitosan oligosaccharide (COS) was investigated in neonatal streptozotocin (STZ)-induced noninsulin-dependent diabetes mellitus rats. The fasting glucose level was reduced by about 19% in diabetic rats after treatment with 0.3% COS. Glucose tolerance was lower in the diabetic group compared with the normal group. After diabetic rats had been treated with 0.3% COS for 4 weeks, glucose tolerance increased significantly versus the diabetic control group, and glucose-inducible insulin expression increased significantly. In addition, fed-triglyceride (TG) levels in diabetic rats drinking 0.3% COS were reduced by 49% compared with those in diabetic control rats. The cholesterol levels of animals treated with COS were reduced by about 10% in fed or fasting conditions versus the corresponding controls, although the difference was not statistically significant. It was found that COS has a TG-lowering effect in diabetic rats, and that COS reduces signs of diabetic cardiomyopathy such as vacuolation of mitochondria and the separation and degeneration of myofibrils. In conclusion, these results indicate that COS can be used as an antidiabetic agent because it increases glucose tolerance and insulin secretion and decreases TG. PMID- 12913259 TI - Comparison of the effects of hange-shashin-to and rikkunshi-to on human plasma calcitonin gene-related peptide and substance P levels. AB - Regarding the gastroprotective function as a neural emergency system, sensory afferent neurons in the gastrointestinal mucosa regulate neuropeptide (calcitonin gene-related peptide (CGRP), substance P, etc.) levels, and those peptides play various physiological roles. To determine whether the pharmacological effects of Hange-shashin-to and Rikkunshi-to on the gastrointestine are due to changes in gastrointestinal mucosa regulatory peptides levels, we investigated the levels of CGRP-like immunoreactive substances (IS) and substance P-IS in plasma from healthy subjects. A single oral administration of Hange-shashin-to caused significant increases in CGRP-IS (40-60 min) and substance P-IS (60-180 min) levels in the plasma compared with the levels induced by a placebo. Rikkunshi-to and a 5.0 g Pinelliae tuber extract had no significant effect on CGRP-IS and substance P-IS levels. Extract of a 2.5 g Zingiberis rhizoma significantly caused increases in CGRP-IS at 40 min and in substance P-IS at 60 min. These results, in comparison with Kampo medicines, might indicate that the pharmacological actions of Hange-shashin-to closely are related to changes in CGRP-IS and substance P-IS levels, while Zingiberis rhizoma partially might participate in those effects of Hange-shashin-to. PMID- 12913260 TI - Oxidation of flavonoids which promote DNA degradation induced by bleomycin-Fe complex. AB - Sixteen flavonoids including quercetin and kaempferol and their relatives were examined for their ability to promote DNA degradation induced by the bleomycin (BLM)-Fe complex. Three hydroxyl groups in the flavonoidal nucleus were proposed as a crucial structural requirement for effectively promoting DNA degradation: 1). the C7-hydroxyl substitution in the A-ring; 2). the C4'-hydroxyl substitution in the B-ring; and 3). the C3-hydroxyl substitution in the C-ring. Flavonoids, which lack even one of these hydroxyl substitutions, showed remarkably diminished activity. There was a good correlation (r=0.920, p<0.001) between activity to promote DNA degradation and oxidizability, which was measured following the Fe(III)-induced oxidation of flavonoids themselves, among the 16 flavonoids. The oxidizability of flavonoids which have the crucial hydroxyl substitutions, was remarkably enhanced in the presence compared with the absence of BLM. On the other hand, the extent of oxidation of flavonoids lacking these substitutions was enhanced little or not at all by BLM. No correlation between the Fe(III)-reducing activity and DNA degradation-promoting activity was found among flavonoids satisfying the crucial structural requirements. Furthermore, the correlation between the extent of oxidation of flavonoids and the Fe(III)-reducing activity was not confirmed among these flavonoids. Therefore, it was suggested that Fe(III)-reducing activity was not the only factor determining DNA degradation promoting activity in flavonoids having the three hydroxyl groups necessary for effectively promoting DNA degradation induced by BLM-Fe complex. PMID- 12913261 TI - Relationship between development of nephrotoxicity and blood concentration of cyclosporine A in bone-marrow transplanted recipients who received the continuous intravenous infusion. AB - This study was undertaken to investigate the relationship between blood concentration of cyclosporine A (CsA), administered intravenously by a 24-h continuous infusion, and drug-induced nephrotoxicity or hepatotoxicity. It was investigated retrospectively in 8 patients who had received an allogeneic bone marrow transplant (BMT). The correlation between daily doses and blood concentration of CsA was not significant. Then, the data of blood concentration of CsA and renal or liver function test result were divided into 5-d periods from the date of transplantation, and the mean value for each period was calculated. The maximum values of blood urea nitrogen (BUN) and serum creatinine (SCr) were consistently observed only after the period when the 5-d mean CsA concentration reached the peak level: the maximum BUN and SCr values were witnessed at Periods 2 to 10 and at Periods 1 to 9, respectively. On the other hand, no consistent correlation was found between the 5-d mean CsA concentrations and liver function test result. We also investigated the relationship between renal function and the cumulative dose or AUC of CsA. The parameters of renal function tests reached peak levels at the cumulative dose of 4000 to 10000 mg and at the cumulative AUC of 280000 to 660000 ng/ml.h. These results suggest that: 1) a deterioration of renal function occurs usually after the peak blood concentration of CsA is attained, and 2) the monitoring of the blood concentration of CsA is useful in predicting renal dysfunction in post-BMT patients. PMID- 12913262 TI - Glucurono- and sulfo-conjugation of kaempferol in rat liver subcellular preparations and cultured hepatocytes. AB - Glucurono- and sulfo-conjugation of kaempferol in rat liver preparations and cultured hepatocytes were studied using high-performance liquid chromatography (HPLC) with two distinctly different elution solvents. Kaempferol glucuronides and sulfates were produced by treating kaempferol with microsomes plus UDPGA or with cytosol plus PAPS, respectively. HPLC analysis of the conjugates revealed one major and three minor glucuronides with solvent A and one sulfate with solvent B. Kaempferol metabolites produced by cultured hepatocytes also consisted of four glucuronides and one minor sulfate, all of which corresponded to their respective in vitro-produced conjugates in the liver subcellular preparations. The relative proportion of kaempferol sulfate accounted for about 9% of the total conjugates in the cultured hepatocytes. The kinetic data on glucurono- and sulfo conjugation of kaempferol by the liver subcellular preparations correlated well with the preferential production of kaempferol glucuronides in the cultured hepatocytes. Glucurono- and sulfo-conjugation of 3-, 5- and 7-OH flavones in the liver subcellular preparations were also kinetically characterized. 7-OH flavone was predominantly conjugated to form a glucuronide compared to 3- and 5-OH flavones. These data suggest that glucuronidation at the 7-OH position on the A ring is a major metabolic pathway of kaempferol in hepatic cells. PMID- 12913263 TI - Inhibitory effects of N-(3,5-dimethoxy-4-n-octyloxycinnamoyl)-N'-(3,4 dimethylphenyl)piperazine (YIC-C8-434), an acyl-CoA:cholesterol O-acyltransferase inhibitor, on cholesterol esterification in the intestine and liver. AB - The effects of an acyl-CoA:cholesterol O-acyltransferase (ACAT) inhibitor, N-(3,5 dimethoxy-4-n-octyloxycinnamoyl)-N'-(3,4-dimethylphenyl)piperazine (YIC-C8-434), on cholesterol esterification in the intestine and liver were investigated in vitro and in vivo. YIC-C8-434 inhibited the formation of cholesteryl [(3)H]oleate from [(3)H]oleic acid and cholesterol both in human colon adenocarcinoma Caco2 cells and in human hepatoma HepG2 cells with IC(50) values of 0.38 and 0.49 microM, respectively. However, it did not influence the incorporation of [(3)H]oleic acid into triacylglycerols and phospholipids. Oral administration of YIC-C8-434 at a dose of 8.3 mg/kg/d inhibited [(14)C]cholesterol absorption by 17% (p<0.01) in rats. YIC-C8-434 also significantly reduced the secretion of very low-density lipoprotein (VLDL) cholesterol from the liver into the plasma at an oral dose of 100 mg/kg/d after an intravenous injection of Triton WR-1339. These results suggest that oral administration of YIC-C8-434 reduces intestinal cholesterol absorption and hepatic VLDL cholesterol secretion by direct inhibition of ACAT in the intestinal epithelium and hepatocytes, respectively. However, the inhibitory action of YIC-C8-434 on cholesterol absorption rather than hepatic cholesterol secretion may play a more important role in its hypocholesterolemic activity, because the effective dose for the former was 12 fold lower than that for the latter. PMID- 12913264 TI - Suppression of free radical-induced DNA strand breaks by linoleic acid and low density lipoprotein in vitro. AB - The results of the present study have shown that unoxidized linoleic acid (LA) and low density lipoprotein (LDL) suppressed free radical-induced supercoiled plasmid DNA strand breaks. Unoxidized LA suppressed DNA strand breaks induced by free radicals generated from hydrogen peroxide/Fe(II) ion, 2'-azobis(2 amidinopropane)hydrochloride (AAPH), and 4-(hydroxymethyl)benzene diazonium salt. Thiobarbituric acid reactive substances (TBARS) of LA were increased on treatment with the radical generators. The intensities of the electron spin resonance (ESR) signals of the spin adducts of the radicals were reduced by unoxidized LA. Although LA hydroperoxide caused DNA strand breaks as has already been shown, its strand breaking activity was observed only at the higher concentrations. Unoxidized LDL inhibited ascorbic acid/Cu(II) ion-, ascorbic acid/Fe(II) ion-, peroxynitrite- and AAPH-induced DNA strand breaks. The TBARS of LDL were increased by treatment with the agents. LDL oxidized with Cu(II) ion did not cause DNA strand breaks. The results indicate that the potency of the free radicals to cause DNA strand breaks was attenuated by the fatty acid and the lipoprotein through lipid peroxidation. PMID- 12913265 TI - Effects of curcuma drugs on vasomotion in isolated rat aorta. AB - The effectiveness of Curcuma drugs against "Oketsu" and the differences in their efficacy were evaluated by examining their vasomotional effects as one index. Since nitric oxide (NO) is the relaxation factor of vascular smooth muscle and also an inhibitor of platelet aggregation in blood vessels, substances showing NO dependent relaxation are thought to be effective in improving Oketsu. In this study, five Curcuma drugs derived from Curcuma longa, C. kwangsiensis, C. phaeocaulis, C. wenyujin, and C. zedoaria were used. Methanol extracts exhibited intense effects on relaxation in rings precontracted by prostaglandin F(2alpha) (PGF(2alpha)) despite pretreatment with and without N(G)-nitro-l-arginine methyl ester (L-NAME) as an inhibitor of NO synthesis. The maximal activities were approximately 80% at 10(-3) g/ml. From these methanol extracts, curcumin and eight sesquiterpenes were isolated. Since all these compounds showed NO independent relaxation effects with almost the same intensities, the relaxation effects of Curcuma drugs can be estimated by the total amounts of curcumin and sesquiterpenes. Polysaccharides, the main constituents of methanol-insoluble compounds of water extracts, in contrast, showed contraction effects; only polysaccharides in C. zedoaria showed NO-dependent relaxation as well as contraction. All water extracts showed relaxation effects as sum of the methanol soluble compounds-induced relaxation and polysaccharides-induced contraction. Therefore, all Curcuma drugs tested in the present study can be effective for vasodilation. Moreover, the drug derived from C. zedoaria has potential to cure Oketsu with its various acting points. PMID- 12913266 TI - Pharmaceutical evaluation of Glycyrrhiza uralensis roots cultivated in eastern Nei-Meng-Gu of China. AB - To clarify the feasibility of medicinal use of the cultivated Glycyrrhiza resources, the equivalency between the G. uralensis roots cultivated in eastern Nei-Meng-Gu of China and medicinal licorice (Glycyrrhizae Radix, Gancao in Chinese and Kanzo in Japanese) was examined. The HPLC fingerprint including glycyrrhizin (GL) of the cultivated roots was similar to that of medicinal Gancao, but different from that of non-medicinal Xinjiang-Gancao (Shinkyo Kanzo in Japanese). Similarity between the cultivated roots and two medicinal Gancao was confirmed quantitatively by hierarchical cluster analysis on the basis of HPLC-7-peak-area data. Moreover, the 4-year-old adventitious roots conformed to the five standards described in the Japanese Pharmacopoeia XIV (JP XIV). The 4 year-old adventitious roots had similar pharmaceutical properties to those of medicinal Dongbei-Gancao (Tohoku Kanzo in Japanese) as determined by examining IgE-mediated triphasic skin reaction in mice and pharmacokinetic profile of glycyrrhetic acid, an anti-allergic metabolite of GL. The present pharmaceutical study suggests that the 4-year-old adventitious roots of G. uralensis cultivated in eastern Nei-Meng-Gu of China are comparable to medicinal Gancao conforming to the JP XIV, and may be a potential medicinal source to compensate for the insufficiency of wild Glycyrrhiza plants caused by collection restriction in China. PMID- 12913267 TI - Effect of glycerol-induced acute renal failure on the pharmacokinetics of lidocaine after transdermal application in rats. AB - In this study, the effect of glycerol-induced acute renal failure (ARF) on the pharmacokinetics of lidocaine after transdermal application was investigated in rats. Microdialysis method was applied in vitro and in vivo to the abdominal skin of rats. After topical application of 1% lidocaine, the cumulative amount of lidocaine permeated through the excised rat abdominal skin showed parallel effect between normal and ARF rats with no significant difference in the in vitro permeability coefficient of lidocaine between them, while area under the plasma concentration versus time curve of lidocaine in ARF rats increased significantly. The protein binding rate of lidocaine in ARF plasma and the blood vessel permeability to muscle tissues, assessed by beta-D-glucopyranosyl fluorescein isothiocyanate-labeled (FITC) albumin, increased significantly. After intravenous infusion of 5 mg/h/kg lidocaine, both of the total body clearance and the volume of distribution of lidocaine in the ARF rats decreased significantly. These results suggested that renal dysfunction did not have any effect on the skin permeability of lidocaine, but might change the plasma protein binding of drug and blood vessel permeability which led to high plasma concentration of lidocaine. PMID- 12913268 TI - Correlation between the physicochemical property of some nonsteroidal anti inflammatory drugs and changes in adenosine triphosphate, glutathione and hemoglobin in rat erythrocytes. AB - This study was conducted to explore the relationship between physicochemical property and toxic effectiveness using rat red blood cells (RBCs). The toxic effectiveness of acid nonsteroidal anti-inflammatory drugs (NSAIDs) was systemically examined by the depletion of intracorpuscular adenosine triphosphate (ATP), glutathione (GSH), and hemoglobin (Hb) at various doses, increased every 5 fmol/RBC. When the RBCs were incubated with NSAIDs, the drugs attained maximum levels within RBC, and the levels were then reduced. The ATP depletion seemed to be observed on the excretion of the drugs prior to the depletions of GSH and Hb. The physicochemical properties of NSAIDs were obtained from QMPRPlus, SMILES code, and CS ChemRaw Ultra. Correlation between their physicochemical properties and their doses for the depletions of ATP, GSH and Hb was performed in comparison with those of the membrane bound enzyme (MBE) inhibiting- and methemoglobin (MHb) generating drugs. The ATP depletion by NSAIDs was correlated with the GSH depletion and intracorpuscular levels of the drugs, but not with the Hb depletion. The GSH depletion was correlated with the Hb depletion and participated in the lipophilicity of the drugs. PMID- 12913270 TI - Purification and characterization of ginsenoside Ra-hydrolyzing beta-D-xylosidase from Bifidobacterium breve K-110, a human intestinal anaerobic bacterium. AB - Beta-D-Xylosidase (EC 3.2.1.37) has been purified from ginsenoside Ra metabolizing Bifidobacterium breve K-110, which was isolated from human intestinal microflora. beta-D-Xylosidase was purified to apparent homogeneity by a combination of ammonium sulfate precipitation, QAE-cellulose, butyl-toyopearl, hydroxyapatit and Q-Sepharose column chromatographies with the final specific activity of 51.8 micromol/min/mg. Molecular weight of beta-D-xylosidase is 49 kDa by SDS-PAGE and gel filtration, which consisted of a single subunit. beta-D Xylosidase showed optimal activity at pH 5.0 and 37 degrees C. The purified enzyme was potently inhibited by PCMS. beta-D-Xylosidase acted to the greatest extent on p-nitrophenyl-beta-D-xylopyranoside, followed by ginsenoside Ra1 and ginsenoside Ra2. This enzyme hydrolyzed xylan to xylose, but did not act on p nitrophenyl-beta-glucopyranoside, p-nitrophenyl-beta-galactopyranoside or p nitrophenyl-beta-D-fucopyranoside. These findings suggest that this is the first reported purification of ginsenoside-hydrolyzing beta-D-xylosidase from an anaerobic Bifidobacterium sp. PMID- 12913269 TI - HPLC analysis of fatty acyl-glycine in the aqueous methanesulfonic acid hydrolysates of N-terminally fatty acylated peptides. AB - Acylation with long-chain fatty acids is a common modification at the N-terminal glycine residues of natural proteins. In this work, we performed HPLC analysis of myristoylglycine (Myr-Gly-OH), palmitoylglycine (Pal-Gly-OH) or lauroylglycine (Lau-Gly-OH), which were produced in the hydrolysates of synthetic Myr-Gly-, Pal Gly-, or Lau-Gly-peptides, respectively, by means of a mild acid hydrolysis in methanesulfonic acid : dioxane : water (2 : 1 : 1) at 60 degrees C for 12 h. Myr Gly-OH, Pal-Gly-OH and Lau-Gly-OH were quite stable under hydrolysis conditions. These fatty acyl-Gly-OH were conveniently detectable at a 20 nmol level by direct reversed-phase HPLC. Thus, mild acid hydrolysis, followed by HPLC analysis of the hydrolysate, provides a simple method of identification of the N-terminal structure of fatty acyl-Gly-peptides. PMID- 12913271 TI - Multinucleation of the sodC-deficient Dictyostelium discoideum. AB - The cellular slime mold Dictyostelium discoideum expresses three genes (sodA, sodB and sodC) encoding the extracellular Cu/Zn superoxide dismutases. Following H(2)O(2) treatment, the expression of sodA and sodB increased while that of sodC decreased. The sodC null strain formed multinucleate cells in a shaking culture. These results suggest that sodC plays a unique role in Dictyostelium discoideum. PMID- 12913272 TI - The role of spinal muscarinic acetylcholine receptors in clonidine-induced anti nociceptive effects in rats. AB - We have examined the effects of intrathecal (i.t.) injection of the muscarinic acetylcholine receptor antagonist atropine on the clonidine-induced nociceptive effect in formalin-induced nociception in rats. The injection of 5% formalin into the hind paw caused biphasic nociceptive responses, and i.t. injection of clonidine inhibited both phases of the nociceptive response in a dose-dependent manner. Pretreatment with atropine (i.t.) only partially inhibited the nociceptive effect of clonidine. These results suggest that the nociceptive effect of clonidine in the rat formalin model may be at least partly mediated by muscarinic acetylcholine receptors in the spinal cord. PMID- 12913273 TI - Cholecystokinin inhibits food intake independent of interleukin-1 beta expression in the brain. AB - Increasing evidence has suggested that cholecystokinin (CCK) is involved in immune-to-brain communication. The afferent vagus nerve is an important component for transmitting peripheral immune signals to the brain, such as those determining interleukin (IL)-1beta expression in the brain and anorexia. In the present study, we investigated whether the anorexic effect of CCK, which also activates the afferent vagus nerve, is mediated via IL-1beta expression in the brain. CCK-8 dose-dependently (8-320 microg/kg, i.p.) inhibited food intake in mice. However, IL-1beta transcripts in the hypothalamus, the hippocampus and the brainstem were not significantly increased after the administration of CCK-8, even at the larger dose of 320 microg/kg. These findings suggest that the CCK induced inhibition of food intake may be independent of IL-1beta production in the brain, and indicate the diverse role of CCK in the regulation of the neuro immune interaction. PMID- 12913274 TI - Assessment of affinities and dissociation potencies of several 5-HT2 antagonists to and from M2 muscarinic receptor in rat heart membranes. AB - The aim of the present study was to investigate the binding affinities and dissociation potencies of several 5-HT(2) antagonists in M(2) muscarinic receptor of rat heart membranes using [(3)H]QNB as a radioligand. The 5-HT(2) antagonists used in this study were sarpogrelate, ketanserin and cyproheptadine. The results showed that sarpogrelate and ketanserin had very weak binding affinities to M(2) muscarinic receptor, whereas cyproheptadine had higher binding affinity to this receptor than the muscarinic receptor antagonist, atropine. All of these three 5 HT(2) antagonists as well as muscarinic receptor antagonists (atropine and pirenzepine) were readily dissociated from M(2) muscarinic receptor in rat heart membranes after washing. Therefore, the findings of the present investigation suggest that the dissociation potencies of neither 5-HT(2) antagonists nor muscarinic antagonists used correlate with their binding affinities to M(2) muscarinic receptors in rat heart. PMID- 12913275 TI - Effect of carotenoid on calcium content and alkaline phosphatase activity in rat femoral tissues in vitro: the unique anabolic effect of beta-cryptoxanthin. AB - The effects of various carotenoids and rutin on calcium content and alkaline phosphatase activity in the femoral-diaphyseal and femoral-metaphyseal tissues of young rats in vitro were investigated. Bone tissues were cultured for 48 h in serum-free Dulbecco's modified Eagle's medium containing either vehicle or a compound (10(-8)-10(-6) M). The presence of beta-cryptoxanthin (10(-7) or 10(-6) M) caused a significant increase in calcium content and alkaline phosphatase activity in the femoral-diaphyseal and femoral-metaphyseal tissues. Also, the lowest concentration (10(-8) M) of beta-cryptoxanthin used caused a significant increase in diaphyseal and metaphyseal calcium content. Lutein (10(-8)-10(-6) M) had no effect on diaphyseal and metaphyseal calcium contents and diaphyseal alkaline phosphatase activity, while 10(-7) and 10(-6) M lutein significantly decreased metaphyseal alkaline phosphatase activity. Lycopene (10(-8)-10(-6) M) or rutin (10(-8)-10(-6) M) did not have a significant effect on bone calcium content and alkaline phosphatase activity. The present study suggests that the carotenoid beta-cryptoxanthin has a unique anabolic effect on bone calcification in vitro. PMID- 12913276 TI - Phospholipase C inhibitors suppress spontaneous mechanical activity of guinea pig urinary bladder smooth muscle. AB - Urinary bladder smooth muscle (UBSM) exhibits spontaneous rhythmic contraction. This spontaneous mechanical activity is generated in the presence of neuronal blockade and thus is myogenic in origin. The spontaneous myogenic contraction of UBSM may be the fundamental determinant of the physiological functions of the urinary bladder to store and excrete urine. Although the mechanisms by which UBSM generates spontaneous contraction have not been completely ascertained, its induction has been suggested to be intimately associated with smooth muscle cell action potentials to enhance extracellular Ca(2+) influx through voltage-gated L type Ca(2+) channels. However, the alteration of membrane electrical activity does not seem to be the exclusive trigger mechanism for the generation of the spontaneous contraction. In the present study, we show that spontaneous mechanical activity of guinea pig UBSM is substantially diminished by an inhibitor of phospholipase C (PLC), U-73122, but is not affected by its inactive form, U-73343. Significant attenuation of the mechanical activity can be also obtained with another PLC inhibitor 2-nitro-4-carboxyphenyl-N,N diphenylcarbamate. Our present findings suggest a significant role for the activation of PLC and subsequent inositol 1,4,5-trisphosphate-induced Ca(2+) release mechanism as an alternative triggering system for inducing spontaneous mechanical activity of UBSM. The present results support the idea that the action potential is not the sole pacemaker mechanism by which spontaneous contraction is induced in UBSM. PMID- 12913277 TI - Upregulation of cellular NF-kappa B activity by alkylating carcinogens in human epidermal keratinocytes. AB - Effect of alkylating carcinogens, i.e., N-nitroso-N-methylurea (NMU) and N nitroso-N-ethylurea (NEU), as well as the simpler alkylating agents, methyl iodide and ethyl iodide, on the activation of NF-kappaB was evaluated in human epidermal squamous cell carcinoma (SCC-13) keratinocytes in order to investigate the possible correlation of cellular NF-kappaB activity with chemical carcinogenesis. The activities of NF-kappaB induced by chemical carcinogens were determined in human SCC-13 keratinocytes transfected with pNF-kappaB-SEAP-NPT plasmid, permitting expression of the secretory alkaline phosphatase (SEAP) reporter gene in response to the NF-kappaB activity and contains the neomycin phosphotransferase (NPT) gene conferring resistance to the geneticin. In this cell-based assay system, all alkylating carcinogens significantly upregulated the cellular NF-kappaB activations in a time- and dose-dependent manner until 72 h, at concentrations of 0.5-5 microM. These results suggest that carcinogenicity by alkylating chemicals may be associated with the modulation of cellular NF-kappaB activity in human skin cells. PMID- 12913278 TI - Cytotoxic activity of steroidal glycosides from solanum plants. AB - Since some Solanum-genera plants have traditionally been used as anti-cancer and anti-herpes agents from olden times, we examined the cytotoxic activity of typical steroidal glycosides with the framework of spirostane, furostane, spirosolane, and pregnane obtained from Solanum plants. Among these steroidal glycosides, the spirostanol glycosides having a beta-lycotetraosyl moiety were the most effective against PC-12 and HCT-116 cell lines. The potency of activity was observed to be decreased in the order of spirostane, furostane, spirosolane, and pregnane type steroid glycosides. It was also suggested that the activity depend on the kind of oligosaccharide moiety and aglycone moiety. PMID- 12913280 TI - Effects of Petroselinum crispum extract on pancreatic B cells and blood glucose of streptozotocin-induced diabetic rats. AB - This study investigated both morphologically and biochemically whether parsley (Petroselinum crispum), which is used as a folk remedy to decrease blood glucose, has any antidiabetic effect on pancreatic B cells of rats. Parsley extract was given to male diabetic rats. In the diabetic group given parsley extract, it was detected that the number of secretory granules and cells in islets and other morphologic changes were not different from the control diabetic group, while the blood glucose levels in the diabetic group given the plant extract were reduced in comparison to the diabetic group. In addition, a decrease was observed in the weight of the control diabetic group and the diabetic group given the plant extract. It is suggested that the plant therapy can provide blood glucose homeostasis and cannot regenerate B cells of the endocrine pancreas. PMID- 12913279 TI - Hepatoprotective dibenzylbutyrolactone lignans of Torreya nucifera against CCl4 induced toxicity in primary cultured rat hepatocytes. AB - Three dibenzylbutyrolactone lignans, (-)-arctigenin, (-)-traxillagenin, and (-) 4'-demethyltraxillagenin, isolated from the bark of Torreya nucifera SIEB. et ZUCC. (Taxaceae) showed significant hepatoprotective activity in primary cultures of rat hepatocytes injured by carbon tetrachloride (CCl(4)). These lignans reduced the release of glutamic pyruvic transaminase into the culture medium from the CCl(4)-injured primary cultures of rat hepatocytes. Further investigation revealed that the three lignans significantly preserved the level of glutathione (GSH) and activities of superoxide dismutase, catalase, glutathione peroxidase, and glutathione reductase in the CCl(4)-injured rat hepatocytes. The lignans also ameliorated lipid peroxidation as demonstrated by a reduction in malondialdehyde related products. Moreover, these lignans significantly attenuated the GSH reduction caused by diethylmaleate which depletes GSH through the formation of stable conjugates. However, these lignans showed no effect on the GSH synthesis inhibited by buthionine sulfoximine. From these results, it can be concluded that arctigenin, traxillagenin, and 4'-demethyltraxillagenin may protect hepatocytes from CCl(4) injury by maintaining the GSH level. PMID- 12913281 TI - Spectrophotometric and fluorimetric determination of hexamine in pure form and its pharmaceutical formulation. AB - Ethyl acetoacetate and methyl acetoacetate have been tested for the determination of hiprex (methenamine hippurate). Both the esters react with hexamine and hiprex forming fluorescent yellow Hantzsch esters. The products show absorption maximum at 370 nm and a fluorescence emission maximum at 465 nm in aqueous n-propanol. Important parameters such as pH, temperature, mole ratio and time of reaction have been optimized. The co-efficient of variation for inter and intra-day precisions were less than 1%. The chromogens show linearity in the concentration ranges of 2.0-35.0 microg x ml(-1) and 0.6-40.0 micro-1) for the spectrometric and fluorimetric determinations of hiprex, respectively. PMID- 12913282 TI - Relative reactivities of histamine and indoleamines with acetaldehyde. AB - Relative reactivities of histamine and indoleamines such as tryptamine, 5 hydroxytryptamine and 5-methoxytryptamine with acetaldehyde (AA) under physiological conditions were investigated. AA was found to have much higher reactivity towards histamine than towards indoleamines. For example, when a reaction mixture of AA (1 mM) and histamine or tryptamine (5 mM) in 0.1 M phosphate buffer (pH 7.4) was incubated at 37 degrees C for 24 h, AA decreased by 11% in the case of tryptamine, while in the case of histamine, it decreased 88%. In addition, the reaction product of AA with histamine was investigated. Mixtures of a fixed amount of histamine (5 mM) and various amounts of AA (1-20 mM) in phosphate buffer (pH 7.4) were incubated for 5 h at 37 degrees C. In all cases, only one product, 4-methylspinaceamine (4-MSPA), was observed. The yield of 4 MSPA was in approximate agreement with the losses of histamine and AA, indicating that the loss of histamine caused by the reaction of AA was quantatively converted to 4-MSPA. These results show that the reaction of AA with histamine easily takes place to produce 4-MSPA in an aqueous medium close to physiological conditions. PMID- 12913283 TI - Estimation of estrogenic and anti-estrogenic activities of some phthalate diesters and monoesters by MCF-7 cell proliferation assay in vitro. AB - Phthalates are man-made chemicals abundantly found in the environment. Estrogenic activities of phthalate di and monoesters were studied by in vitro assay of human breast cancer MCF-7 cell proliferation. Since phthalate monoesters are formed from diesters by degradation and are found in the environment, we selected some phthalate monoesters in addition to diesters. Among 19 compounds tested, dicyclohexyl phthalate (DCHP), di(2-ethylhexyl) phthalate (DEHP) and butyl benzyl phthalate (BBP) were found to have estrogenic activities, all of which were completely suppressed by the addition of pure anti-estrogen ICI 182780. DCHP stimulated cell proliferation with maximal cell yield at 5 x 10(-5) M. Its estrogenic potency was approximately 1700000 times less than that of 17beta estradiol. DEHP and BBP stimulated cell proliferation only slightly at >10(-3) M. No other phthalate diesters or monoesters tested were estrogenic. Anti-estrogenic activities were also examined by estimating the suppression of cell proliferation in the presence of 10(-11) M 17beta-estradiol. Mono-n-pentyl phthalate (MPP), monocyclohexyl phthalate (MCHP), monobenzyl phthalate (MBZP), monoisopropyl phthalate (MIPrP) and BBP were suggested to have anti-estrogenic activities at higher than 10(-4) M. Among commonly used phthalate esters and those with related structures, some were found to be estrogenic and others were anti-estrogenic in vitro. PMID- 12913284 TI - Antibody to soluble 1,3/1,6-beta-D-glucan, SCG in sera of naive DBA/2 mice. AB - A branched beta-glucan from Sparassis crispa (SCG) is a major 6-branched 1,3-beta D-glucan showing antitumor activity. In the present study, we examined the anti SCG antibody in naive mice by ELISA. Using SCG coated plate, sera of naive DBA/1 and DBA/2 mice contained significantly higher titers of antibody than other strains of mice. Anti-SCG Ab titers of each DBA/1 and DBA/2 mice were significantly varied. Using various polysaccharide-coated plate, sera of DBA/2 mice also reacted with a beta-glucan from Candida spp. (CSBG) having 1,3-beta and 1,6-beta-glucosidic linkages. The SCG specific immunoglobulin (Ig) M but G was detected in sera. The reactivity of sera to coated SCG was neutralized by adding soluble SCG and CSBG as competitor. These results suggested that DBA/1 and DBA/2 strains carry specific and unique immunological characteristics to branched 1,3 /1,6-beta-glucan. PMID- 12913285 TI - New potent antioxidative hydroxyflavanones produced with Aspergillus saitoi from flavanone glycoside in citrus fruit. AB - Potent antioxidative hydroxyflavanones were produced with Aspergillus saitoi from hesperidin or naringin, which are flavanone glycosides in citrus fruit with weak antioxidative activity. The hydroxyflavanone produced from hesperidin was identified as 8-hydroxyhesperetin (8-HHE), a novel substance, and those from naringin were identified as carthamidin (6-hydroxynaringenin) and isocarthamidin (8-hydroxynaringenin) by FAB-MS, 1H-NMR and 13C-NMR analyses. The antioxidative activity of these hydroxyflavanones was examined by using the free radical scavenging system of 1,1-diphenyl-2-picrylhydrazyl (DPPH) and the methyl linoleate oxidation system. The hydroxyflavanones (8-HHE, carthamidin, and isocarthamidin) exhibited stronger activity than the flavanone glycosides (hesperidin or naringin) and their aglycones (hesperetin or naringenin). The activity of 8-HHE and isocarthamidin was comparable to that of alpha-tocopherol, and that of carthamidin was weaker than that of isocarthamidin. The hydroxyflavanones, which were hydroxylated on A ring of flavanone by Aspergillus saitoi, were obtained as potent antioxidants. PMID- 12913286 TI - Anti-obesity effect of Dioscorea nipponica Makino with lipase-inhibitory activity in rodents. AB - In the process of screening for pancreatic lipase inhibitors, which could be used as an anti-obesity measure, the methanol extract of Dioscorea nipponica Makino powder (DP) appeared to have potent inhibitory activity against porcine pancreatic lipase with an IC50 value of 5-10 microg/ml, where the enzyme activity was assayed by using 4-methylumbelliferyl oleate as a substrate. Further purification of active components present in the herb generated dioscin that belongs to the saponin family. Dioscin and its aglycone, diosgenin, both suppressed the time-dependent increase of blood triacylglycerol level when orally injected with corn oil to mice, suggesting their inhibitory potential against fat absorption. Sprague-Dawley rats fed on a high-fat diet containing 5% Dioscorea nipponica Makino and 40% beef tallow gained significantly less body weight and adipose tissue than control animals fed on a high-fat diet alone during an 8-week experimental period (P<0.05). PMID- 12913287 TI - High-cholesterol diets induce changes in lipid composition of rat erythrocyte membrane including decrease in cholesterol, increase in alpha-tocopherol and changes in fatty acids of phospholipids. AB - Effects of high dietary cholesterol on erythrocyte membrane lipids were studied. Feeding rats with a diet containing 0.5% cholesterol and 0.15% sodium cholate for two weeks induced changes in erythrocyte membrane lipids including a decrease in cholesterol, an increase in alpha-tocopherol (alpha-Toc) and changes in the fatty acid composition of phospholipids. Oleic acid and linoleic acid increased, while arachidonic acid decreased in phosphatidylcholine. Saturated fatty acids decreased and unsaturated fatty acids increased in phosphatidylethanolamine. Almost the same changes in membrane lipids were also noted after six weeks of feeding rats with the diet. A diet containing 0.5% cholesterol but without sodium cholate caused a decrease in erythrocyte cholesterol and an increase in erythrocyte alpha-Toc after two weeks of feeding, as compared to the basal diet, indicating that high dietary cholesterol, but not sodium cholate, was responsible for these changes in the erythrocyte membrane. PMID- 12913288 TI - Production of galactinol from sucrose by plant enzymes. AB - Galactinol, 1-O-(alpha-D-galactopyranosyl)-myo-inositol, was produced from sucrose as a starting material. UDP-Glc was prepared with sucrose and UDP using sucrose synthase partially purified from sweet potato roots. Then, the UDP-Glc was converted to UDP-Gal using yeast UDP-Gal 4-epimerase from a commercial source. Finally, galactinol was produced from the UDP-Gal and myo-inositol using galactinol synthase partially purified from cucumber leaves. The product was identified as galactinol by the retention times of HPLC, alpha-galactosidase digestion, and NMR spectrometry. PMID- 12913289 TI - Either soluble or plastidic expression of recombinant protoporphyrinogen oxidase modulates tetrapyrrole biosynthesis and photosynthetic efficiency in transgenic rice. AB - Protoporphyrinogen oxidase (Protox) is the last shared enzyme of the porphyrin pathway. As a continuation of our previous work in which the transgenic rice plants expressing the Bacillus subtilis Protox in the cytoplasm or the plastid showed resistance to diphenyl ether herbicide, this study was undertaken to identify the effects of tertapyrrole biosynthesis in these transgenic rice plants. The transgenic plants either targeted into plastids or expressed in cytoplasm showed higher Protox activity than wild-type plants did. Photosynthetic activity, measured as a quantum yield of photosystem II, was slightly higher in transgenic plants than in wild-type plants, but chlorophyll contents were not significantly different between transgenic and wild-type plants. As for porphyrin biosynthesis, both cytoplasm-expressed and plastid-targeted transgenic plants showed increased synthesis of aminolevulinic acid, Mg-Proto IX, and protoheme in comparison to wild-type plants whereas synthesis of protoporphyrin IX was similar for wild-type and transgenic plants. These results indicate that either cytoplasm or plastid expression of B. subtilis Protox in rice can upregulate the porphyrin pathway leading to increase in photosynthetic efficiency in plants. PMID- 12913290 TI - Effects of high concentrations of inorganic salts on swarming ability in fluorescent pseudomonas strains. AB - We did tests using swarm plates, to examine the effects of various salts and their concentrations on the chemotaxis of fluorescent Pseudomonas strains. As a result, we found that the swarming ability of the Pseudomonas strains was inhibited by high concentrations of Ca2+. The growth of the strains was not affected at the high concentration of Ca2+, but the cells grown in swarm agar under the condition were extended in the filaments. Most of the cells had reached 10 microm to 40 microm in length. Such cell elongation was not observed with salts other than calcium salts. A significant correlation between the cell elongation and the decrease of swarming ability by the high concentration of Ca2+ was observed. PMID- 12913291 TI - Purification and characterization of thermostable alpha-galactosidase from Ganoderma lucidum. AB - Alpha-galactosidase was purified from a fresh fruiting body of Ganoderma lucidum by precipitation with ammonium sulfate and column chromatographies with DEAE Sephadex and Con A-Sepharose. The purified enzyme was homogeneous on polyacrylamide gel electrophoresis. Its N-terminal amino acid sequence was similar to that of Mortierella vinacea alpha-galactosidase. The molecular mass of the enzyme was about 56 kDa by SDS-polyacrylamide gel electrophoresis, and about 249 kDa by gel filtration column chromatography. The optimum pH and temperature were 6.0 and 70 degrees C, respectively. The enzyme was fully stable to heating at 70 degrees C for 30 min. It hydrolyzed p-nitrophenyl-alpha-D-galactopyranoside (Km=0.4 mM) but hydrolyzed little o-nitrophenyl-alpha-D-galactopyranoside. It also hydrolyzed melibiose, raffinose, and stachyose. The enzyme catalyzed the transgalactosylation reaction which synthesized melibiose. The product was confirmed by various analyses. PMID- 12913292 TI - Active stress during compression testing of various foods measured using a multiple-point sheet sensor. AB - Using a multiple-point sheet sensor (MSS), load and contact area were directly measured for compression of four different foods. The MSS provided temporal and spatial changes in stress applied on the sample surface during the testing. The sum of load value detected by the MSS corresponded to the load measured by a universal testing machine during the compression. The contact area between a flat probe and food surface varied with the variety of foods even though under a small strain, and increased as compression strain increased. The active stress, that is, the load divided by the contact area, was different from conventional stress, that is, the load divided by the initial cross-sectional area. The value of active stress provided a better explanation of textural characteristics of food, because texture is often sensed under a large deformation and mixed assessment of mechanical and geometrical properties. PMID- 12913293 TI - Stimulation of glucose uptake in muscle cells by prolonged treatment with scriptide, a histone deacetylase inhibitor. AB - Glucose incorporation is regulated mainly by GLUT4 in skeletal muscles. Here we report that treatment of L6 myotubes with scriptide, a hydroxamic acid-based histone deacetylase (HDAC) inhibitor, stimulated 2-deoxyglucose uptake. The effect appeared only after 24 hr, resulting in 2.4-fold glucose uptake at treatment day 6. Scriptide acted synergistically with insulin, indicating it stimulated a distinct pathway from the insulin signaling pathway. It was not observed in undifferentiated myoblasts or 3T3-L1 adipocytes, suggesting a muscle specific effect of scriptide. A five-carbon chain and hydroxamic acid, essential for histone deacetylase inhibition, were indispensable for this effect, and trichostatin A stimulated glucose uptake as well. Scriptide increased the cellular content of GLUT4, and induced GLUT4 translocation, but GLUT4 mRNA level did not change, indicating scriptide functions posttranslationally. Our results indicated a novel function for HDAC inhibitors of increasing GLUT4 content and its translocation in muscle cells, resulting in stimulation of glucose uptake. PMID- 12913294 TI - Amino acid residue substitution at T-cell determinant-flanking sites in beta lactoglobulin modulates antigen presentation to T cells through subtle conformational change. AB - We compared T-cell responses to regions in residues 21-40 of A and B variants of bovine milk beta-lactoglobulin (beta-LG) that vary by two different amino acid residues at 64 and 118. Results showed that T cells from C57/BL6 and C3H/HeN mice immunized with peptide 21-40 or BALB/c mice immunized with peptide 21-32 or 25-40 responded more vigorously to beta-LG B than to beta-LG A. This difference in response to 25-40 in BALB/c mice was not observed when beta-LGs B and A were denatured, suggesting that the conformation difference affects display of the determinant 25-40. Reactivity of anti-beta-LG monoclonal antibodies and molecular modeling using molecular dynamics calculations revealed subtle differences in the three-dimensional structure of these two variants. Furthermore, substitution of two amino acid residues at sites distant from the T-cell determinant induced differential determinant display on antigen-presenting cells, possibly due to subtle conformational changes in beta-LG. PMID- 12913295 TI - Genomic structure and 5'-flanking sequences of rat N acetylglucosaminyltransferase I gene and regulatory role of its transcriptional diversity. AB - It has been known that mouse, rat, and human N-acetylglucosaminyltransferase I (GnT-I) genes produce at least two transcripts, which differ in their 5' untranslated region (5'-UTR) length, and the longer transcript is preferentially expressed in brains. However, the physiological meaning of this brain-specific expression pattern was unknown. We cloned the rat GnT-I gene and analyzed its structure. It consisted of five exons, and four of them coded only 5'-UTRs. A putative NF-kappaB binding site was found in the 5'-flanking sequence for the transcript that was previously shown to be induced by inflammation. The unusually long 5'-UTR of the major GnT-I transcript in rat brain was shown to inhibit protein production from the following coding sequence in COS7 cells. Comparison of the GnT-I protein/mRNA ratio in rat brain and liver showed that GnT-I mRNA in the brain was translated 3.8-times less efficiently than in the liver. Implications are discussed of these results in regulation of GnT-I expression in rat brain. PMID- 12913296 TI - Interaction between elongation factors 1beta and 1gamma from Bombyx mori silk gland. AB - Elongation factor 1 (EF-1) from the silk gland of Bombyx mori consists of four subunits: alpha (51 kDa), beta (26 kDa), gamma (49 kDa), and delta (33 kDa). The EF-1alpha subunit catalyzes the binding of aminoacyl-tRNA to the ribosome concomitant with the hydrolysis of GTP. The EF-1alpha-bound GDP is then exchanged for GTP by the EF-1betagammadelta complex. To facilitate analysis of the roles of the individual EF-1beta, gamma, and delta subunits in GDP/GTP exchange on EF 1alpha, we cloned the cDNAs for these subunits and expressed them in Escherichia coli. EF-1beta, EF-1gamma, and the carboxyl-terminal half of EF-1delta were expressed, purified, and examined for protein:protein interactions by gel filtration chromatography and by a quartz-crystal microbalance method. An 80-kDa species containing EF-1beta and gamma subunits in a 1:1 molar ratio was detected by gel filtration. A higher molecular weight species containing an excess of EF 1gamma relative to EF-1beta was also detected. The amino-terminal region of EF 1beta (amino acid residues 1-129) was sufficient for binding to EF-1gamma. The carboxyl-terminal half of EF-1delta did not appear to form a complex with EF 1gamma. PMID- 12913297 TI - Synthesis of glyceroyl beta-N-acetyllactosaminide and its derivatives through a condensation reaction by cellulase. AB - A condensation reaction between N-acetyllactosamine and glycerol was directly catalyzed by using a commercially available cellulase preparation from Trichoderma reesei. 1-O-beta-N-Acetyllactosaminyl-(R, S)-glycerols (1) were readily synthesized in a 5% yield based on the N-acetyllactosamine added and conveniently isolated by two-step column chromatographies. The use of a partially purified enzyme increased 2.3-fold the yield of 1, compared to that of the crude enzyme containing beta-D-galactosidase activity. When various alkanols (n:2-4) were used in the condensation reaction, the corresponding alkyl beta-N acetyllactosaminides were obtained in yields of 0.3-1.1% of the desired compounds. PMID- 12913298 TI - Volatilization of mercury by an iron oxidation enzyme system in a highly mercury resistant Acidithiobacillus ferrooxidans strain MON-1. AB - A highly mercury-resistant strain Acidithiobacillus ferrooxidans MON-1, was isolated from a culture of a moderately mercury-resistant strain, A. ferrooxidans SUG 2-2 (previously described as Thiobacillus ferrooxidans SUG 2-2), by successive cultivation and isolation of the latter strain in a Fe2+ medium with increased amounts of Hg2+ from 6 microM to 20 microM. The original stain SUG 2-2 grew in a Fe2+ medium containing 6 microM Hg2+ with a lag time of 22 days, but could not grow in a Fe2+ medium containing 10 microM Hg2+. In contrast, strain MON-1 could grow in a Fe2+ medium containing 20 microM Hg2+ with a lag time of 2 days and the ability of strain MON-1 to grow rapidly in a Fe2+ medium containing 20 microM Hg2+ was maintained stably after the strain was cultured many times in a Fe2+ medium without Hg2+. A similar level of NADPH-dependent mercury reductase activity was observed in cell extracts from strains SUG 2-2 and MON-1. By contrast, the amounts of mercury volatilized for 3 h from the reaction mixture containing 7 microM Hg2+ using a Fe(2+)-dependent mercury volatilization enzyme system were 5.6 nmol for SUG 2-2 and 67.5 nmol for MON-1, respectively, indicating that a marked increase of Fe(2+)-dependent mercury volatilization activity conferred on strain MON-1 the ability to grow rapidly in a Fe2+ medium containing 20 microM Hg2+. Iron oxidizing activities, 2,3,5,6-tetramethyl-p phenylenediamine (TMPD) oxidizing activities and cytochrome c oxidase activities of strains SUG 2-2 and MON-1 were 26.3 and 41.9 microl O2 uptake/mg/min, 15.6 and 25.0 microl O2 uptake/mg/min, and 2.1 and 6.1 mU/mg, respectively. These results indicate that among components of the iron oxidation enzyme system, especially cytochrome c oxidase activity, increased by the acquisition of further mercury resistance in strain MON-1. Mercury volatilized by the Fe(2+)-dependent mercury volatilization enzyme system of strain MON-1 was strongly inhibited by 1.0 mM sodium cyanide, but was not by 50 nM rotenone, 5 microM 2-n-heptyl-4-hydroxy quinoline-N-oxide (HQNO), 0.5 microM antimycin A, or 0.5 microM myxothiazol, indicating that cytochrome c oxidase plays a crucial role in mercury volatilization of strain MON-1 in the presence of Fe2+. PMID- 12913299 TI - Fourth 3D structure of the chitosan molecule: conformation of chitosan in its salts with medical organic acids having a phenyl group. AB - Chitosan salts with two medical organic acids having phenyl groups (salicylic and gentisic acids) exhibited fiber diffraction patterns of a new type of crystal which does not compare with known types I and II. The crystals, called type III salts, showed a fiber repeat of 2.550 nm and a meridional reflection at the 5th layer line. These results coupled with a conformational analysis indicate the chain conformation of chitosan with the salts to be a 5/3 helix, this helix differing from those of type I (an extended two-fold helix) and type II (a relaxed two-fold helix or a 4/1 helix). The fiber patterns of all the type III salts were similar. This observation has also been found with type II salts and is an indication that the acid ions are not arranged in regular positions in the crystals. A comparison of solid-state 13C-NMR spectra of the gentisic acid salt and the aspirin salt, which could not be crystallized, suggests that, in the latter salt, the chitosan molecules also formed a 5/3 helix. PMID- 12913300 TI - Deactivation of gibberellin by 2-oxidation during germination of photoblastic lettuce seeds. AB - Gibberellin (GA) plays an important role in the induction of germination of photoblastic lettuce (Lactuca sativa L. cv. Grand Rapids) seeds. We have previously shown that gene expression of a GA 3-oxidase (Ls3h1) increased after a red light treatment, resulting in an increase in the endogenous content of GA1, bioactive GA. Since the metabolism of GAs is also important for determining the endogenous levels of bioactive GAs, cDNAs encoding GA 2-oxidases (LsGA2ox1 and LsGA2ox2, for L. sativa GA 2-oxidase), which catalyze the deactivation of GAs, were isolated from lettuce seeds to investigate the regulation of these genes by light. An expression analysis shows that the mRNA levels of both enzymes was not markedly altered under different light conditions during germination. However, the amount of LsGA2ox2 transcripts was decreased to approximately half the level by red light. This reduction might play a role in the increase in GA1 level by red light in the lettuce seeds. PMID- 12913301 TI - Synthesis of the four components of the female sex pheromone of the painted apple moth, Teia anartoides. AB - Four pheromone components of the female painted apple moth (Teia anartoides), an Australian insect pest, were synthesized. These were (Z)-6-henicosen-11-one (1), (6Z, 8E)-6,8-henicosadien-11-one (2), (Z)-cis-9,10-epoxy-6-henicosene (3), and (Z)-cis-9,10-epoxy-6-icosene (4). 2-Dodecanone was converted to 1 and 2, and both the enantiomers of 3 and 4 were synthesized from the enantiomers of 4-tert butyldimethylsilyloxy-cis-2,3-epoxy-1-butanol. PMID- 12913302 TI - Structural studies and antifungal activity of unique polyene amides, clathrynamide A and three new derivatives, from a marine sponge, Psammoclemma sp. AB - Known polyene amide, clathrynamide A, and three novel related metabolites, debromoclathrynamide A, (4E,6E)-debromoclathrynamide A and (6E)-clathrynamide A, were isolated from an Okinawan marine sponge, Psammoclemma sp. The absolute stereochemistry of clathrynamide A was determined to be 3R,12R by the modified Mosher method. The structures of the three new compounds were elucidated by spectroscopic analyses as debromo derivatives and geometrical isomers. Their antifungal activities were also evaluated with a phytopathogenic fungus, indicating that both the bromine atom and double-bond geometry affected the activity. PMID- 12913303 TI - Development of an effective sample preparation method for the proteome analysis of body fluids using 2-D gel electrophoresis. AB - A sample preparation is still the most critical step in two-dimensional electrophoresis (2-DE) and should be optimized for each type of sample. In this study, a protein extraction method from body fluids was developed using a combined centrifugal filter device and a sample treating buffer. When plasma, amniotic fluid, urine, and tear were tested with this method, the recovery of protein reached almost 90% and high-quality separation of 2-DE gel was obtained. PMID- 12913304 TI - Quinone hemiacetal formation from protocatechuic acid during the DPPH radical scavenging reaction. AB - Protocatechuic acid was rapidly converted to protocatechuquinone 3-methyl hemiacetal and protocatechuquinone during the reaction with DPPH radical in methanol. The structure of the acetal was determined by comparing the NMR data with those of an authentic compound prepared by (diacetoxy)iodobenzene oxidation of protocatechuic acid. PMID- 12913305 TI - A novel isoindoline, porritoxin sulfonic acid, from Alternaria porri and the structure-phytotoxicity correlation of its related compounds. AB - Novel zinniol-related compound 3, named porritoxin sulfonic acid, with an isoindoline skeleton was isolated from the culture liquid of Alternaria porri. The structure was determined to be 2-(2"-sulfoethyl)-4-methoxy-5-methyl-6-(3' methyl-2'-butenyloxy)-2,3-dihydro-1H-isoindol-1-one. The phytotoxic activities of three isoindolines (1-3) were evaluated in a seedling-growth assay against stone leek and lettuce. PMID- 12913306 TI - Conjugated linoleic acid (CLA) inhibits fatty acid synthetase activity in vitro. AB - This paper describes the in vitro effect of conjugated linoleic acid (CLA) on fatty acid biosynthesis. Among the rat liver enzymes involved in fatty acid biosynthesis, fatty acid synthetase (FAS) showed the largest activity fluctuation with the types of fatty acids. Of the fatty acids, CLA was the most potent inhibitor of FAS, and the 9c, 11t-rather than the 10t, 12c-isomer showed greater inhibition. CLA also significantly lowered the incorporation of [14C]-acetate into phospholipid in breast cancer cells, supporting the view that CLA inhibits fatty acid biosynthesis through the interaction with FAS. PMID- 12913307 TI - Inhibitory effects of resveratrol derivatives from dipterocarpaceae plants on tyrosinase activity. AB - Stilbene derivatives, which are resveratrol (3,4',5-trihydroxy-trans-stilbene) oligomers ranging from monomer to tetramer, isolated from Dipterocarpaceae plants were tested for their inhibitory effects against murine tyrosinase activity. The structure-activity relationships obtained in this study suggest that the double bond in the stilbene skeleton is critical for the inhibition, and also that molecular size is important for inhibitory potency. PMID- 12913308 TI - Degradation of car engine base oil by Rhodococcus sp. NDKK48 and Gordonia sp. NDKY76A. AB - Two microorganisms (NDKK48 and NDKY76A) that degrade long-chain cyclic alkanes (c alkanes) were isolated from soil samples. Strains NDKK48 and NDKY76A were identified as Rhodococcus sp. and Gordonia sp., respectively. Both strains used not only normal alkane (n-alkane) but also c-alkane as a sole carbon and energy source, and the strains degraded more than 27% of car engine base oil (1% addition). PMID- 12913309 TI - Isolation and identification of sodium 2-propenyl thiosulfate from boiled garlic (Allium sativum) that oxidizes canine erythrocytes. AB - Sodium 2-propenyl thiosulfate was identified in boiled garlic (Allium sativum). When canine erythrocytes were incubated with sodium 2-propenyl thiosulfate, the methemoglobin concentration and Heinz body percentage in erythrocytes were both increased, indicating that the compound induced oxidative damage in canine erythrocytes. It seems that this compound is one of the causative agents of garlic-induced hemolysis in dogs. PMID- 12913311 TI - Conversion of the cleavage specificity of subtilisin YaB on oxidized insulin chains to an elastase-like specificity by replacement of Gly124 with Ala. AB - Replacement of Gly124 on the S1 pocket of subtilisin YaB with Ala changed the cleavage pattern on oxidized insulin B-chain from the subtilisin type to the elastase type. The initial cleavage site in the B-chain shifted from L15-Y16 for wild-type YaB to A14-L15 for the G124A mutant. Upon complete hydrolysis with the G124A mutant, four of the six major cleavage sites on the B-chain were identical to porcine pancreatic elastase cleavage sites. PMID- 12913310 TI - New plant growth promoters, repraesentins A, B and C, from Lactarius repraesentaneus. AB - Three new plant growth regulatory sesquiterpenoids were isolated from the Lactarius repraesentaneus fungus. Their structures were elucidated to be a protoilludene sesquiterpene, namely repraesentin A (1), and two related sesquiterpenes, namely repraesentins B (2) and C (3). Compounds 1-3 showed promotion activities toward the radicle elongation of lettuce seedlings by 136%, 118% and 184% at 67 ppm, respectively. PMID- 12913312 TI - Transfer of pro-R hydrogen from NADH to dihydroxyacetonephosphate by sn-glycerol 1-phosphate dehydrogenase from the archaeon Methanothermobacter thermautotrophicus. AB - sn-Glycerol-1-phosphate dehydrogenase is responsible for the formation of the sn glycerol-1-phosphate backbone of archaeal lipids. [4-3H]NADH that had 3H at the R side was produced from [4-3H]NAD and glucose with glucose dehydrogenase (a pro-S type enzyme). The 3H of this [4-3H]NADH was transferred to dihydroxyacetonephosphate during the sn-glycerol-1-phosphate dehydrogenase reaction. On the contrary, in a similar reaction using alcohol dehydrogenase (a pro-R type enzyme), 3H was not incorporated into glycerophosphate. These results confirmed a prediction of the tertiary structure of sn-glycerol-1-phosphate dehydrogenase by homology modeling. PMID- 12913313 TI - Purification and characterization of purple acid phosphatase PAP1 from dry powder of sweet potato. AB - Purple acid phosphatase (PAP) was purified from sweet potato dry powder, which is used as a food additive. Spectrometric and enzymatic analyses, and analysis of the amino-terminal sequence indicated that the purified purple acid phosphatase was PAP1. High activity in neutral and acidic conditions, broad substrate specificity, and good thermal stability of PAP1 suggest the possibility of practical applications of PAP1. PMID- 12913314 TI - Identification and classification of two-component systems that affect rpoS expression in Escherichia coli. AB - The rpoS-encoded sigmaS subunit of RNA polymerase regulates the expression of stationary phase and stress response genes in Escherichia coli. Recent study of our DNA microarray analysis suggested that the rpoS expression is affected by multiple two-component systems. In this study, we identified two-component-system mutants in which the rpoS expression increased. The regulatory manner of the systems on rpoS expression is suggested. PMID- 12913315 TI - Identification of the lantibiotic nisin Q, a new natural nisin variant produced by Lactococcus lactis 61-14 isolated from a river in Japan. AB - Lactococcus lactis 61-14 isolated from river water produced a bacteriocin active against a wide range of Gram-positive bacteria. N-terminal amino acid sequencing, mass spectral analysis of the purified bacteriocin, and genetic analysis using nisin-specific primers showed that the bacteriocin was a new natural nisin variant, termed nisin Q. Nisin Q and nisin A differ in four amino acids in the mature peptide and two in the leader sequence. PMID- 12913316 TI - Enterostatin (VPDPR) and its peptide fragment DPR reduce serum cholesterol levels after oral administration in mice. AB - We found that enterostatin (VPDPR), an anorexigenic peptide for a high-fat diet, significantly reduces serum cholesterol levels after oral administration of 100 mg/kg for 3 days in mice fed a high cholesterol-cholic acid diet. DPR, a peptide fragment of VPDPR, also had hypocholesterolemic activity at a dose of 50 mg/kg. Food intake was not suppressed under these dietary conditions. Fecal excretion of cholesterol and bile acids was increased significantly by both VPDPR and DPR. Interestingly, DPR induced hypocholesterolemic effects just two hours after a single oral administration at a dose of 100 mg/kg. PMID- 12913317 TI - Anti-angiogenic activity of tocotrienol. AB - The anti-angiogenic property of vitamin E compounds, with particular emphasis on tocotrienol, has been investigated in vitro. Tocotrienol, but not tocopherol, inhibited both the proliferation and tube formation of bovine aortic endothelial cells, with delta-tocotrienol appearing the highest activity. Also, delta tocotrienol reduced the vascular endothelial growth factor-stimulated tube formation by human umbilical vein endothelial cells. Our findings suggest that tocotrienol has potential use as a therapeutic dietary supplement for minimizing tumor angiogenesis. PMID- 12913318 TI - Influence of cycloversions induced by head tilt on scanning laser polarimetry parameters. AB - OBJECTIVE: To study the influence of cycloversions induced by head tilt on parameters of scanning laser polarimetry. METHODS: Five left and five right eyes of 9 normal subjects were measured in seven different head tilt positions by one examiner using a scanning laser polarimeter (Nerve Fiber Analyzer, NFA II, LDT Inc., San Diego, Calif., USA). Images were acquired for the following head tilt positions: upright, 10, 20, 30 degrees to the right, and 10, 20, 30 degrees to the left. For each position five to eight images of the peripapillary area were recorded. Objective cycloversions were determined trigonometrically on the fundus pictures provided by the scanning laser ophthalmoscope. Parameters acquired for different head positions were compared using a Friedmann's two-way analysis of variance. RESULTS: Average objective cyclodeviations were 5.9 degrees for the 10 degrees head tilts, 11.2 degrees for the 20 degrees head tilts, and 18.9 degrees for the 30 degrees head tilts. Head tilts to the right in left eyes and to the left in right eyes (both of them corresponding to an incycloposition) induced a significant increase in several parameters (p < 0.05). Head tilts to the left in left eyes and to the right in right eyes (both corresponding to an excycloposition) showed no influence on parameters (p > 0.05). CONCLUSIONS: Ocular incycloposition has a significant influence on several scanning laser polarimetry parameters. Therefore, evaluation is limited in patients having an incyclorotation of the nonfixating eye with the head in an upright position or in patients having a head tilt inducing an incyclorotation. PMID- 12913319 TI - Tumour regression of uveal melanoma after ruthenium-106 brachytherapy or stereotactic radiotherapy with gamma knife or linear accelerator. AB - PURPOSE: This study assesses differences in relative tumour regression and internal acoustic reflectivity after 3 methods of radiotherapy for uveal melanoma: (1) brachytherapy with ruthenium-106 radioactive plaques (RU), (2) fractionated high-dose gamma knife stereotactic irradiation in 2-3 fractions (GK) or (3) fractionated linear-accelerator-based stereotactic teletherapy in 5 fractions (Linac). METHODS: Ultrasound measurements of tumour thickness and internal reflectivity were performed with standardised A scan pre-operatively and 3, 6, 9, 12, 18, 24 and 36 months postoperatively. Of 211 patients included in the study, 111 had a complete 3-year follow-up (RU: 41, GK: 37, Linac: 33). Differences in tumour thickness and internal reflectivity were assessed with analysis of variance, and post hoc multiple comparisons were calculated with Tukey's honestly significant difference test. RESULTS: Local tumour control was excellent with all 3 methods (>93%). At 36 months, relative tumour height reduction was 69, 50 and 30% after RU, GK and Linac, respectively. In all 3 treatment groups, internal reflectivity increased from about 30% initially to 60 70% 3 years after treatment. CONCLUSION: Brachytherapy with ruthenium-106 plaques results in a faster tumour regression as compared to teletherapy with gamma knife or Linac. Internal reflectivity increases comparably in all 3 groups. Besides tumour growth arrest, increasing internal reflectivity is considered as an important factor indicating successful treatment. PMID- 12913320 TI - Automated analysis of eye tracking movements. AB - OBJECTIVE: To present a quick algorithm to automatically analyze the raw data acquired by a photo-oculography (POG) system. METHODS: We developed a simple algorithm for POG data analysis based on an extrapolation of missing values due to blinking and on exclusion of outliers using the robust mean and standard deviation. RESULTS: POG curves of 4 children aged between 1.5 and 7 years are shown before and after automatic analysis. After applying our algorithm, the curves are much smoother. CONCLUSION: Our algorithm allows a quick data analysis and will help to better interpret and analyze POG data. PMID- 12913321 TI - Phacoemulsification and intraocular lens implantation for angle closure glaucoma after the relief of pupillary block. AB - PURPOSE: We studied the surgical outcomes of phacoemulsification and intraocular lens (IOL) implantation for cataract and/or uncontrolled intraocular pressure (IOP) in eyes with angle closure glaucoma. SETTING: Department of Ophthalmology, National Nagasaki Medical Center, Nagasaki, Japan. METHODS: Eighteen eyes from 15 patients after laser iridotomy (17 eyes) or peripheral iridectomy (1 eye) had undergone surgery and were studied. We used an iris retractor in 7 eyes due to insufficient mydriasis and a capsular tension ring in 2 eyes due to phacodonesis during the operation. The patients were followed up for at least 6 months (13.8 +/- 7.2 months; range: 6-36 months). RESULTS: The mean IOP significantly decreased from 17.4 +/- 8.1 to 13.5 +/- 3.3 mm Hg at 6 months after surgery. The IOP was below 21 mm Hg in all eyes. The visual acuity was not worsened in any eyes and became better than 2 Snellen lines in 14 eyes. The corneal endothelial cell count decreased from 2,365 +/- 517 to 1,960 +/- 661/mm2 (18.3 +/- 17.2%). CONCLUSIONS: Phacoemulsification and IOL implantation is useful in IOP control for angle closure glaucoma after relief of pupillary block. However, we should take care of operative complications because of a shallow anterior chamber, poor mydriasis and zonular weakness. PMID- 12913322 TI - Effects of extended-wear soft contact lenses on the ocular surface and central corneal thickness. AB - PURPOSE: New contact lens materials with high oxygen permeability are said to be convenient for the extended-wear (EW) schedule. We evaluated the short-term effect of EW soft contact lens usage on ocular surface and central corneal thickness. PATIENTS AND METHODS: Fifty-two eyes of 26 patients were included in the study. Sixteen of the patients used the high oxygen-permeable lenses for a 30 day extended-wear schedule (group I). Ten patients used the lenses on a daily wear basis (DW, group II). All patients were evaluated for visual acuity and central corneal thickness by corneal and conjunctival biomicroscopy, Schirmer and tear break-up time (BUT) tests. Impression cytology was performed in the superior/temporal, inferior/nasal quadrants of the conjunctiva, 1 mm from the limbus, with cellulose acetate filter paper. The specimens were graded under light microscopy according to Nelson's classification. RESULTS: Three patients in the EW group were excluded from the study as they discontinued lens usage due to slight discomfort and hyperemia. Biomicroscopic findings and tear function tests indicated no increased pathological results. Central corneal thickness decreased from 554 +/- 9.55 to 546 +/- 9.30 microm (mean +/- SEM) in the EW group (p = 0.002), whereas no significant change occurred in the DW group. Cytological evaluation of the EW group showed an 88% increase in metaplasia (p = 0.007), especially close to the limbal areas and in the superior quadrants; this ratio was 30% in the DW group. Snake-like chromatin changes were found to be increased, from 4 to 6 patients, in the EW group while it remained the same in the DW group (p > 0.05). Individual comparisons of the initial and final pathological conditions of the ocular surfaces in the two groups showed that there was an increase in pathology in the EW group while there was no significant difference in the DW group. CONCLUSIONS: On a 30-day extended-wear basis with high oxygen permeable lenses, the possible development and increase of conjunctival cytologic changes secondary to mechanical trauma is one of the main remaining problems. Conjunctival and related immunologic pathologies should not be overlooked when prescribing an extended-wear lens schedule. PMID- 12913323 TI - Protective effects of pentoxifylline in retinal ischemia/reperfusion injury. AB - We studied the effect of pentoxifylline on retinal lipid peroxidation and histopathologic changes due to ischemia/reperfusion (I/R). A total of 15 pigmented male guinea pigs were divided into 3 equal groups as control, sham and treatment groups. After application of high intraocular pressure for 90 min for the induction of retinal ischemia, 24-hour reperfusion was established in the sham and treatment groups. In the treatment and sham groups, either 45 mg/kg of pentoxifylline or saline was given 3 times at 8-hour intervals. Biochemical assay and histopathologic evaluation were performed on one randomly selected eye of each animal which was enucleated at the end of the reperfusion period, and retinal malondialdehyde (MDA) levels and thickness of the retinal tissue were determined for each group. The mean MDA level of the sham group was significantly higher versus the control and treatment groups (p < 0.001). When compared with the control group, the mean MDA level of the treatment group was slightly higher, but the difference was not statistically significant (p > 0.05). In comparison with the control group there was a significant increase in the thickness of the retina in the sham group (p < 0.0001), and no significant difference was found in the retinal thickness of the treatment group (p > 0.05). Pentoxifylline might have a preventive effect on the I/R injury of the retina. PMID- 12913324 TI - Possible involvement of nitric oxide in the progression of diabetic retinopathy. AB - Abnormal nitric oxide (NO) synthesis has been implicated in the pathogenesis of diabetes mellitus. The aim of our study was to elucidate the relationship between the stages of diabetic retinopathy (DR) and the NO levels in aqueous humor and plasma. Using the chemiluminescence assay, we measured the concentrations of NO in aqueous humor and plasma samples obtained during intraocular surgery from 45 diabetic patients and 19 nondiabetic cataract patients. The patients with diabetes were classified into 4 groups: proliferative DR (PDR) with active neovascularization (active PDR; 9 cases), PDR with quiescent neovascularization (regressed PDR; 6 cases), background DR (BDR; 16 cases) and no DR (14 cases). We found that the aqueous NO levels (mean +/- SE) of the active PDR group (83.2 +/- 13.9 microM) were significantly higher than those of the BDR group (45.8 +/- 6.0 microM, p = 0.049) and the diabetics without DR (33.3 +/- 5.2 microM, p = 0.011), and, although not statistically significantly, they were also higher than those of the regressed PDR group (52.1 +/- 10.3 microM, p = 0.224). However, no significant differences were observed between any of the diabetic subgroups in the plasma NO levels (p = 0.345). We therefore concluded that NO present in the ocular tissues may play important roles in the progression of DR. PMID- 12913325 TI - The effect of blood glucose regulation on retinal nerve fiber layer thickness in diabetic patients. AB - PURPOSE: To evaluate the effect of blood glucose (BG) regulation on the retinal nerve fiber layer (RNFL) in diabetic patients by using a scanning laser polarimeter (NFA-GDx). METHODS: We prospectively assessed RNFL thickness in diabetic patients and an age-matched control group. Patients without diabetic retinopathy, with BG >250 mg/dl, HbA1c >8%, fructosamine >285 micromol/l and triglyceride >200 mg/dl were included in the study. RNFL assessment was performed before and after metabolic regulation of diabetes. Symmetry, superior maximum, ellipse modulation and the average thickness variables of NFA-GDx were used for the assessment. Mann-Whitney U and Wilcoxon tests were used for the statistical analysis. RESULTS: A total of 40 diabetic patients were included in the study and a repeat RNFL examination could be performed in 22 of them following regulation of BG levels. None of the GDx variables were significantly different between pre- and postregulation measurements (p > 0.05, Wilcoxon test). The mean superior maximum, ellipse modulation and average thickness values of the diabetic group were significantly lower than the control group (p < 0.05, Mann-Whitney U-test). CONCLUSIONS: Poor metabolic control of diabetes mellitus adversely affects the thickness of RNFL and this effect does not seem to be acute since it was not reversed by short-term BG regulation. This issue needs to be kept in mind when assessing glaucomatous progress in diabetic patients. PMID- 12913326 TI - Mitotropic compounds for the treatment of age-related macular degeneration. The metabolic approach and a pilot study. AB - Recent histopathologic studies have shown that mitochondria and peroxisomes of the retinal pigment epithelium may play a central role in the pathophysiology of age-related macular degeneration (AMD). We supposed that compounds which improve mitochondrial functions (mitotropic compounds) may show beneficial effects in preventing AMD. Fourteen patients affected by early AMD were treated with a mixture containing acetyl-L-carnitine (ALC), polyunsaturated fatty acids (PUFAs), coenzyme Q10 (CoQ10) and vitamin E, while an equal number of age- and sex-matched patients affected by early AMD were treated with vitamin E only. Recovery time after macular photostress, foveal sensitivity and mean defect in the visual field as well as blood lipid levels were recorded at the beginning and after 3, 6, 9, 12 and 24 months of follow-up. In the treated group, all the visual functions showed slight improvement which was evident after 3 months of treatment and remained nearly stationary by the end of 24 months. The same tests in the control group showed slow worsening. The divergence between treated and control groups became more marked with time, but the difference was not significant at any time of the follow-up. These findings suggest that the blend of ALC, PUFA, CoQ10 and vitamin E may improve retinal functions in early AMD. PMID- 12913327 TI - Association of interleukin 1beta and receptor antagonist gene polymorphisms with primary open-angle glaucoma. AB - PURPOSE: Genetic factors are known to play a role in the etiology of glaucoma. More specifically, the role of the immune system is highly suspected. We evaluated the association between 2 polymorphisms in the interleukin (IL) 1beta gene (IL-1 promoter 511, IL-1 exon 5) and 1 polymorphism in the IL-1 gene receptor antagonist (IL-1Ra) intron 2 with primary open-angle glaucoma (POAG). PATIENTS AND METHODS: Fifty-eight POAG patients and 105 healthy volunteers were enrolled in this study. Analysis based on polymerase chain reaction was used to resolve the 2 IL-1beta polymorphisms and the IL-1Ra intron 2 polymorphism. RESULTS: There were significant differences in the distribution of the IL-1beta exon 5 polymorphism between the POAG patients and the control subjects (p < 0.05). The E2 allele of IL-1beta exon 5 was more frequently found in POAG patients than in healthy patients (odds ratio: 4.224, 95% confidence interval = 1.135-15.717). The distributions of the other 2 polymorphisms, IL-1beta promoter 511 and IL-1Ra intron 2, were not significantly different between the POAG patients and the healthy control group. CONCLUSIONS: The frequency of the E2 allele of the IL-1beta exon 5 polymorphism was high in POAG patients. Therefore the E2 allele can be used as a marker to predict or search for the genetic causes of glaucoma in Chinese POAG patients. Furthermore, we have concluded that the other 2 polymorphisms (IL-1 promoter 511 and IL-1Ra) are not helpful in predicting Chinese POAG. PMID- 12913328 TI - Decreased activation of the lateral geniculate nucleus in a patient with anisometropic amblyopia demonstrated by functional magnetic resonance imaging. AB - Although postmortem morphological changes in the lateral geniculate nucleus (LGN) have been reported in human amblyopia, LGN function during monocular viewing by amblyopic eyes has not been documented in humans. We used functional magnetic resonance imaging (fMRI) to study monocular visual activation of the LGN in a patient with anisometropic amblyopia. Four normal subjects, a patient with optic neuritis and a patient with anisometropic amblyopia were studied with fMRI at 1.5 T during monocular checkerboard stimulation. Activated areas in the LGN and visual cortex were identified after data processing (motion correction and spatial normalization) with SPM99. In the 4 normal subjects, comparable activation of the LGN and visual cortex was obtained by stimulation of either the right or left eye. In the patient with unilateral optic neuritis, activation of the LGN and visual cortex was markedly decreased when the affected eye was stimulated. Similarly, decreased activation of the LGN as well as the visual cortex by the affected eye was demonstrated in the patient with anisometropic amblyopia. Our preliminary results suggest that activation of the LGN is diminished during monocular viewing by affected eyes in anisometropic amblyopia. fMRI appears to be a feasible method to study LGN activity in human amblyopia. PMID- 12913329 TI - Pseudodoubling of the optic disc: a colour Doppler imaging study. AB - Pseudodoubling of the optic disc is a rare clinical presentation. In these cases it is necessary to exclude retinal coloboma or atrophy following vascular or infectious diseases. We present a case of pseudodoubling of the optic disc in a woman with type 2 diabetes and arterial hypertension. Ophthalmoscopic examination of the fundus showed a disc-like lesion in the right eye and a diabetic retinopathy in the left eye. The lesion was evaluated with fluorescein angiography, neuroradiological and colour Doppler imaging investigations. Colour Doppler imaging confirmed the angiographic findings of anomalous vascularisation of the pseudopapilla and provides an analysis of the choroidal vessel anastomosis between the optic disc and the retinal lesion, revealing that the pseudodoubling in this patient was the result of a chorioretinal coloboma. PMID- 12913330 TI - Reversion of 'early worsening' of diabetic retinopathy by deliberate restoration of poor metabolic control. AB - Acutely lowering long-standing severe hyperglycaemia can trigger progression ('early worsening') of diabetic retinopathy, most likely by up-regulation of the circulating insulin-like growth factor 1 (IGF-1). This condition, also called 'florid retinopathy', rarely responds to standard laser coagulation treatment. In this retrospective report, 2 young patients with type 2 diabetes are described, in whom deliberate restoration of poor diabetes control reduced the serum IGF-1 levels and improved 'early worsened' diabetic retinopathy. PMID- 12913331 TI - Subretinal hemorrhage displacement: a new surgical technique without iatrogenic retinal break or evacuation procedures. PMID- 12913332 TI - Highly effective non-viral gene transfer into vascular smooth muscle cells of cultured resistance arteries demonstrated by genetic inhibition of sphingosine-1 phosphate-induced vasoconstriction. AB - The linkage of vascular genes to specific functions will lead to a better understanding of cardiovascular pathophysiology. We developed an experimental model that enables the introduction of one or multiple gene(s) into vascular smooth muscle cells (VSMCs) of isolated resistance arteries. Exposure of the arteries to a green fluorescent protein (GFP)-encoding plasmid in combination with the transfectant Effectene((R)) for 20 h resulted in the expression of GFP in virtually all VSMCs in the arterial wall at fully preserved vascular function. For functional validation of the model, plasmids encoding the specific RhoA inhibitors C3 transferase or N19RhoA were transfected. In subsequent functional tests, inhibition of RhoA-dependent constriction induced by sphingosine-1 phosphate was similar to that in arteries treated with exogenous C3 transferase protein or the Rho kinase inhibitor Y27632. Responses to norepinephrine remained unaffected. This novel transfection technique enables gene function to be assessed in direct conjunction with signalling pathways in vascular tissue and provides, therefore, a new tool for microvascular proteomics. PMID- 12913333 TI - Collagen biomechanics in cerebral arteries and bifurcations assessed by polarizing microscopy. AB - Collagen is the main matrix protein of the artery wall. We have used the known correlation between collagen birefringence and its mechanical properties to assess the wall structural integrity in brain arteries and their bifurcation regions, which are the sites of formation of saccular aneurysms. Segments of 28 brain arteries, including bifurcations, were pressure fixed and sectioned in one of three orthogonal planes. Measurements were taken by polarizing microscopy of the birefringence of collagen fibers at the apex of bifurcations and in the main layers of the artery wall - adventitia, media and intima. Dimensional data were obtained of the layers in order to estimate wall properties. Along the apex of the flow divider we measured a narrow band of collagen (birefringence 30% higher than the adjacent adventitia) providing strength and stiffness in that region. There is a thin cell-free outer layer of the tunica media (mean thickness 11 microm) comprised of densely packed coaligned collagen with high birefringence. From the fiber birefringence and directional alignment of the individual layers we calculated that the adventitia contributes about one third of circumferential and almost all of longitudinal strength of intracranial arteries. PMID- 12913334 TI - Effect of isoflurane on skin-pressure-induced vasodilation. AB - Since general anesthesia has been shown to attenuate endothelium-dependent vasodilation, it was of interest to verify whether general anesthesia would modify skin vasodilation in response to local pressure application, which is endothelium dependent. To study the effect of general anesthesia on pressure induced vasodilation development, we examined the effects of low- and high-dose isoflurane. Skin blood flow was measured by laser Doppler flowmetry during 11.1 Pa s(-1) increases in locally applied pressure in anesthetized rats treated with low or high doses of isoflurane. Following the administration of low doses of isoflurane, skin blood flow increased from baseline, with increasing local pressure application (+37 +/- 10% at 2.0 kPa). The increase in skin blood flow was absent in rats treated with high doses (-20 +/- 5% at 2.0 kPa), even when the anesthesia-induced hypotension was corrected by gelofusine infusion (-20 +/- 10% at 2.0 kPa). Whereas sodium-nitroprusside-induced vasodilation developed following low and high doses of isoflurane, acetylcholine-induced vasodilation was impaired with high doses compared to low doses. These data show that pressure induced vasodilation is abolished with high doses of anesthetics. It is not the anesthesia-induced hypotension, but the depth of anesthesia, which can lead to the disappearance of pressure-induced vasodilation by an alteration in endothelial function. PMID- 12913335 TI - Non-viable cervico-isthmic pregnancy: the importance of an accurate sonographic diagnosis to preserve fertility. AB - BACKGROUND: Cervico-isthmic pregnancy is a rare occurrence and until the use of current ultrasonographic techniques was associated with a disastrous outcome for women desiring to maintain their fertility. CASE REPORT: A 39-year-old woman was diagnosed at 12 weeks of amenorrhoea with an intra-uterine non-viable pregnancy and a low implantation of the gestational sac. Medical management of this situation with a regimen of intravaginal misoprostol was unsuccessful. At introduction of a cervical expander severe vaginal bleeding ensued, leading to an emergency curettage followed by a life-saving hysterectomy. Pathologic examination confirmed the peroperative impression of an isthmic implantation of the gestational sac. CONCLUSION: Early ultrasonographic diagnosis is essential to make conservative management of cervico-isthmic pregnancy possible. PMID- 12913336 TI - Local anesthetics inhibit uterine activity in vitro. Possible application on preterm labor prevention and treatment. AB - OBJECTIVE: Analysis of the effects of local anesthetics upon the gravid uterus. METHODS: Isolated strips of pregnant-rat myometrium were placed within tissue baths. Muscular activity was then recorded by a force transducer, at cumulative concentrations of either procaine, lidocaine, or ropivacaine. RESULTS: The effects of myometrial activity were non-monotonic. At lower concentrations, an increase in contraction frequency was noted, concomitant with a depression in amplitude. As the concentrations increased, both amplitude and frequency were depressed until, eventually, activity was abolished. CONCLUSIONS: Local anesthetics administered in situ consistently inhibit or completely arrest overall myometrial activity. These agents may prove clinically useful in preterm labor management, if administered locally to the myometrium via suitable slow release drug delivery systems, warranting further trials in vivo. PMID- 12913337 TI - Intrauterine repair of gastroschisis in fetal rabbits. AB - OBJECTIVE: Infants with gastroschisis (GS) still face severe morbidity. Prenatal closure may prevent gastrointestinal organ damage, but intrauterine GS repair (GSR) has not been established yet. METHODS: In New Zealand White rabbits we developed and compared GS versus GSR: creation of GS was achieved by hysterotomy, right-sided laparotomy of the fetus and pressure on the abdominal wall to provoke evisceration. GSR was accomplished by careful reposition of eviscerated organs and a running suture of the fetal abdominal wall. For study purposes, 18 animals were divided equally into 3 groups: GS, GS with GSR after 2 h, and unmanipulated controls (C). Vitality was assessed by echocardiography. After 5 h all animals were sacrificed. RESULTS: GSR inflicted no increased mortality, because all fetuses survived GS or GS with GSR. All fetuses with GS demonstrated significant evisceration of abdominal organs. In contrast, the abdominal wall of the fetuses from GSR was intact. CONCLUSION: The present animal model demonstrated the technical feasibility and success of an intrauterine repair of GS for the first time. However, further long-term studies (leaving GS and GSR in utero for several days) will be necessary to compare survival rates and intestinal injury, motility or absorption. The clinical application of GSR in utero remains a vision so far. PMID- 12913338 TI - Discordant fetal growth patterns in monochorionic twin pregnancies described by simple algebraic relations. AB - OBJECTIVE: To test whether twin-twin transfusion syndrome (TTTS) and non-TTTS cases that were suspect but not confirmatory for developing the syndrome (non TTTS-with-symptoms) have discordant fetal growth patterns that correlate with our previously derived algebraic relations. METHODS: In 25 monochorionic twin pregnancies, fetal growth was determined by standard ultrasonography. The difference between estimated fetal weights (dEFW) as well as the difference divided by the average of the two weights, the difference average ratio (DAR), were fitted to the predicted trends of discordant fetal growth for TTTS and non TTTS pregnancies. The best fits were compared with the clinical data. RESULTS: Out of 13 TTTS cases, dEFW analysis correctly correlated with 8 (62%) and DAR analysis correctly with 10 (77%). Out of 12 non-TTTS-with-symptoms cases, dEFW analysis correctly correlated with 7 (58%) and DAR analysis correctly with 9 (75%). If TTTS correlated best, dEFW analysis was correct in 8/12 (67%), and DAR analysis in 10/11 (91%) cases. If non-TTTS correlated best, dEFW analysis was correct in 7/9 (78%), and DAR analysis in 9/12 (75%). The likelihood ratios of TTTS and non-TTTS were 1.9 and 0.26, respectively, with dEFW analysis, and 9.2 and 0.31 with DAR analysis. CONCLUSION: The simple algebraic relations derived to identify trends of fetal discordant growth show evidence that clinical TTTS and non-TTTS manifestations are quantifiable. The relations may contribute to future risk stratification in monochorionic twin pregnancies, e.g., by prospectively distinguishing between cases that will develop TTTS and those that will not, despite presenting with symptoms. PMID- 12913339 TI - Identification of fetal nucleated red blood cells in the maternal circulation during pregnancy using anti-hemoglobin-epsilon antibody. AB - AIM: To investigate the use of anti-hemoglobin-epsilon antibody in order to identify fetal cells in the maternal circulation during pregnancy. MATERIALS AND METHODS: 48 blood samples were obtained from pregnant women, 26 in the 1st trimester and 22 in the 2nd trimester. Magnetic activated cell sorting was used for fetal cell enrichment followed by immunophenotyping with a monoclonal antibody against hemoglobin-epsilon. FISH with X, Y and 21 chromosome-specific probes was performed in 29 cases. RESULTS: The mean number of epsilon-positive cells was 9.2 (range 2-23) in the 1st trimester, 4.8 (range 3-13) in the 2nd trimester and 22 (range 15-28) in pregnancies with Down syndrome. No significant difference was noted in the number of epsilon-positive nucleated red blood cells (NRBCs) isolated from carriers and noncarriers of beta-thalassemia. FISH analysis was successful in 24 cases. In 4 cases with known male fetuses, an average of 4.7 epsilon-positive cells showed a Y signal. In 4 cases with Down syndrome, all epsilon-positive cells showed 3 signals for chromosome 21. CONCLUSION: Anti hemoglobin-epsilon antibody has increased specificity for fetal NRBCs and should be preferentially used to improve noninvasive prenatal diagnosis of chromosome abnormalities from fetal cells in maternal blood. PMID- 12913340 TI - Chorioamniotic membrane separation following open fetal surgery: pregnancy outcome. AB - OBJECTIVE: To review the incidence of posthysterotomy chorioamniotic membrane separation and delivery outcome following open fetal surgery [myelomeningocele (MMC); cystic adenomatoid malformation (CCAM); congenital diaphragmatic hernia (CDH); sacrococcygeal teratoma (SCT)]. STUDY DESIGN: Retrospective review of a maternal population undergoing open fetal surgery at a single tertiary level program (1998-2001) following the initiation of close postoperative ultrasound follow-up for membrane separation. Onset of membrane separation was coded as not present (NP), immediate (<2 weeks) or delayed (>2 weeks) from day of surgery. RESULTS: Fifty-three charts were reviewed: MMC 43, CCAM 7, CDH 1, and SCT 2. In the MMC group there were 26 NP, 8 immediate, and 9 delayed. Preterm labor occurred in 4 patients with only 2 having had membrane separation. Risk of membrane separation is increased for surgery done at less than 23 weeks gestation (p < 0.005). Delay from MMC surgery to delivery was 11.0, 9.8, 12.0 weeks for NP, immediate, and delay, respectively. In the MMC group, there were 3 neonatal deaths (NND) at 9, 9, and 21 days post surgery (PROM/PTL; chorioamnionitis, PROM/PTL, respectively). No membrane separation was present in the CCAM, CDH, and SCT cases. CONCLUSIONS: (1) Membrane separation was significantly more likely to occur if surgery was performed prior to 23 weeks. (2) Membrane separation post hysterotomy (17/50 = 34%) may be associated with an increased risk of PROM but not delivery before 30 weeks gestation. (3) Delivery prior to 33 weeks gestation for MMC groups was 12/43 (28%) with 3 NND (7%). (4) Elective delivery at 36-37 weeks gestation was possible for 43% of the fetal surgery population. PMID- 12913341 TI - Maternal serum and amniotic fluid hydroxyproline levels in neural tube defects. AB - OBJECTIVE: The present study was aimed to evaluate the alterations of maternal serum and amniotic fluid OH- proline levels in neural tube defects (NTD). METHODS: 38 patients and 23 control subjects were selected from the patients followed up by the Departments of Genetics and Antenatal Care, Zekai Tahir Burak Women's Health, Education and Research Hospital. Amnion fluid and blood samples were synchronously obtained from NTD and control subjects. OH-proline levels were detected spectrophotometrically. RESULTS: Amniotic fluid OH-proline levels were found to be increased in NTD patients compared to control subjects (p < 0.05). There was no statistically significant difference in serum OH-proline levels between the groups. CONCLUSION: The results showed an increase in amniotic fluid OH-proline levels in NTD compared to the control group. It may be suggested that the increased OH-proline levels reflect increased collagen turnover in NTD patients. Indeed, amniotic fluid OH-proline levels may be a candidate as an additional diagnostic parameter in NTD. PMID- 12913342 TI - Prenatal diagnosis of fetal intracranial hemorrhage at 25 weeks of gestation. AB - OBJECTIVE: To determine whether magnetic resonance imaging (MRI) can give further information for the prenatal diagnosis of intracranial hemorrhage, when ultrasound (US) is not conclusive. METHODS: We report on a fetus in the 26th week of gestation with a grade-III hemorrhage in the right cerebral hemisphere, which was suspected on US examination and specified by MRI examination in a patient with pre-eclampsia. RESULTS: Doppler measurement of the umbilical artery, which showed the absence of end diastolic velocity, was indicative of placental insufficiency. The MRI specified the findings found by US examination and confirmed the poor outcome of the fetus. Labor was induced by prostaglandins in the 27th week of gestation. Postmortem examination showed a female fetus with a grade-III intracranial hemorrhage in the center of the right hemisphere with a diameter of 4 cm. A hemorrhage was also found in the subarachnoid space and in the occipital region of the left hemisphere. CONCLUSION: We suggest that fetuses at high risk of developing hypoxia should be sonographically evaluated and MRI should be considered when intracranial echogenicities are seen. PMID- 12913344 TI - Epinephrine inhibits tracheal occlusion induced lung growth in fetal sheep. AB - OBJECTIVE: Although the mechanisms responsible for lung growth following tracheal occlusion (TO) are not fully understood, lung fluid accumulation is a requirement for growth to occur. It is known that betamimetics such as epinephrine (Epi), terbutaline, and ritodrine inhibit fetal lung fluid production late in gestation. We hypothesized that continuous infusion of Epi would have a detrimental effect on lung growth following TO in the fetal sheep model. METHODS: Twenty fetal lambs were divided into four groups. Group 1: no TO without Epi (n = 4); group 2: TO without Epi (n = 4); group 3: no TO with Epi (n = 5), and group 4: TO with Epi (n = 7). TO was performed on days 130-134 of gestation (full term = 145 days). In groups 3 and 4, Epi was infused into the fetus at a rate of 2 microg/min using a miniosmotic pump implanted subcutaneously at the time of TO. The fetuses were sacrificed after 4 days, and lung volume (LV, ml/kg), drained lung fluid (LF, ml/kg), wet lung weight/body weight ratio (LW/BW, %), whole right-lung dry weight per body weight ratio (dRLW/BW, g/kg), volume density of lung parenchyma (v(Vp)), and right-lung DNA and protein contents were compared among the four groups by one-way Anova. RESULTS: LW/BW and dRLW/BW of group 4 (3.91 +/- 0.52 and 2.10 +/- 0.28, mean +/- SD) were significantly lower than those of group 2 (5.18 +/- 0.57 and 2.67 +/- 0.15) and were not statistically significantly different from those of group 1 or 3. LV, LF, V(Vp), and DNA and protein contents all showed a similar trend. CONCLUSIONS: Continuous infusion of Epi to the fetus results in less fluid accumulation within the TO lungs and abolishes significant lung growth after TO in the late-gestation fetal lamb. Drugs affecting lung fluid secretion may have a major impact on TO-induced lung growth. PMID- 12913343 TI - Prenatal diagnosis in beta-thalassemia: an Indian experience. AB - OBJECTIVES: Thalassemia is the most common single gene disorder and is widely distributed in Asian Indians with an average prevalence rate of 4%, with a high prevalence among Sindhis, Punjabis, Gujratis and Bengalis. Prevention and control of beta-thalassemia disease require the accurate diagnosis of carriers and proper genetic counseling. METHOD: Prenatal diagnosis can be performed in the first or second trimester of pregnancy by DNA analysis using polymerase chain reaction. Since there are 17 mutations as well as rare ones causing beta-thalassemia in Asian Indians, the point mutation detection by reverse dot blot (RDB) allele specific oligonucleotide hybridization for common mutations along with the amplification refractory mutation system (ARMS) technique was developed for prenatal diagnosis. Maternal contamination of fetal DNA was ruled out by the variable number of tandem repeat analysis using apolipoprotein B site. RESULTS: Using both techniques (RDB and ARMS) we were able to offer complete diagnosis in 53 pregnancies. On molecular analysis 23% were found to be normal, 48.0% were carriers, and 29.0% were affected with beta-thalassemia. Parents were counseled to continue the pregnancy when the fetuses were either normal or had traits whereas in the case of an affected fetus, the parents opted for termination of the pregnancy. CONCLUSION: Prenatal diagnosis of beta-thalassemia by the RDB or ARMS technique can prevent the birth of an affected child in developing countries in which beta-thalassemia is quite prevalent. PMID- 12913345 TI - Antenatal evaluation of an encephalocele in a dizygotic twin pregnancy using fast magnetic resonance imaging. AB - We report a case of an encephalocele in a dizygotic twin pregnancy, following ovulatory induction. In the involved fetus, an abnormal shadow like an encapsulated-solid tumor located on the occiput was found by routine maternal transabdominal ultrasonography at 17 weeks of gestation. The parents did not accept induced abortion because of the presence of another fetus with no abnormality on ultrasonography. At 35 weeks of gestation, transabdominal ultrasound examination showed a large occipital cyst, composed of protrusive fetal brain and cerebrospinal fluid. Fast-scanning magnetic resonance imaging delineated more clearly the inside of the abnormal lesion and thus allowed confirmation of the putative diagnosis of fetal encephalocele during pregnancy. Surgical report was possible in this case, and the patient had no severe physical or neurological abnormalities 10 months after birth. Since the prognosis appears to depend primarily on how prominent the brain tissue is inside the herniated sac, this approach had benefit for clinical decision making. PMID- 12913346 TI - Body stalk anomaly diagnosed in the 2nd trimester. AB - We examined a 20-year-old primigravida by ultrasound at 21 weeks of gestation during routine anomaly scanning. Several fetal deformities were demonstrated: the upper part of the body was in the amniotic cavity, while the lower part was in the celomic cavity. A large abdominal wall defect was detected with herniation of the liver and the intestine. The limbs were deformed, the spine had severe kyphoscoliosis, and the umbilical cord was very short. These findings were suggestive of a body stalk anomaly. Termination of the pregnancy was offered and decided by the parents. The pathology report confirmed the ultrasonographic diagnosis. PMID- 12913347 TI - Do microbiological factors account for poor pregnancy outcome among unmarried pregnant women in Poland? AB - OBJECTIVE: Being unmarried is a well-known risk factor for poor pregnancy outcome such as preterm delivery and intrauterine growth restriction. The aim of this prospective study was to assess the prevalence and risk of bacterial vaginosis (BV) and selected bacteria isolated from the lower genital tract and to determine the socioeconomic and microbiological characteristics that might be responsible for poor pregnancy outcome observed among unmarried pregnant women. METHODS: The study population comprised 196 pregnant women attending 10 randomly selected outpatient maternity units in the Lodz region, central Poland. Cervicovaginal samples were obtained between 8 and 16 weeks of gestation. Based on Spiegel's criteria, gram-stained vaginal smears were examined for BV and the BV-associated flora was sought by culture. To evaluate the risk factors, relative risk ratios were calculated using EPI INFO software. RESULTS: Among 196 pregnant women, 40 (20.4%) were unmarried. BV was diagnosed among 55 (28.1%) women studied. In the univariate analysis, unmarried pregnant women were characterized by younger age, primary educational level, poor economic situation and excessive smoking during pregnancy, as compared to married women. The unmarried status was a borderline risk factor for BV (OR = 1.83, 95% CI 0.94-4.9) after adjustment for age, smoking and education. An analysis of the microbiological culture from the lower genital tract revealed that unmarried pregnant women had a higher risk for several types of pathological microflora, as compared to married women. However, this finding was significant only for Mycoplasma hominis. The independent risk factors of M. hominis were the young age of the subject and a low concentration of Lactobacillus spp. CONCLUSIONS: The observed socioeconomic, demographic and microbiological differences between unmarried and married women could be responsible for the poor pregnancy outcome among unmarried pregnant women in Poland. Unmarried pregnant women should be covered by comprehensive medical care even before pregnancy. Further studies taking into account the role of psychological stress, patterns of sexual behavior and substance abuse during pregnancy could help identify the factors responsible for adverse pregnancy outcome among unmarried pregnant women. PMID- 12913348 TI - Postmortem validation of imaging-derived formulas for prediction of fetal lung volume. AB - OBJECTIVE: To determine the validity of existing imaging-derived formulas for predicting the fetal lung volume (FLV). METHODS: In a consecutive series of postmortem lungs without pulmonary anomalies, the observed FLV (FLV(obs)) after inflation was correlated with individual fetal variables used in imaging-derived formulas. In addition, FLV(obs) was correlated with the predicted FLV calculated according to these same formulas. RESULTS: Postmortem FLV(obs) showed a strong correlation with estimated fetal body weight, biparietal diameter, and head circumference (r > 0.9). The correlation of FLV(obs) with gestational age, femur length, and liver weight was less strong (0.8 < r < 0.9). The correlation was strongest for midgestation fetuses (between 22 and 32 weeks' gestation). The predicted FLV calculated using formulas based on fetal body weight, biparietal diameter, and head circumference showed the strongest correlation with the actual FLV(obs) (r > 0.9). CONCLUSIONS: Postmortem FLV(obs) strongly correlates with fetal body weight, biparietal diameter, and head circumference, especially in midgestation fetuses. Regression formulas based on these fetal variables provide the most accurate prediction of FLV. PMID- 12913349 TI - Effect of alpha-fetoprotein and derived peptides on insulin- and estrogen-induced fetotoxicity. AB - Both insulin and estrogen are well recognized as growth-promoting substances at physiological concentrations, but they function as teratogens at high doses. Both agents can affect alterations in fetal and maternal serum human alpha-fetoprotein (HAFP) levels during pregnancy. In the present study, we have employed animal models of both insulin and estrogen fetotoxicity and teratogenicity in order to study the growth-regulatory properties of HAFP and its derived peptides (HAFP/PEP). We report here the effects of HAFP/PEP on fetotoxicity, congenital malformations, and growth retardation in developing chick and murine fetuses. In the insulin model, HAFP/PEP were effective in reducing both fetal mortality and anatomic anomalies, with the result that growth-retarded fetuses were produced. With HAFP/PEP treatment, fetal demise was reduced by as much as 73 and 63% in murine and chick fetuses, respectively, while fetal anomalies were diminished by 50% during chick development. Genebank searches of identity/similarity in a HAFP/PEP fragment identified matches with a number of proteins associated with glucose, pH, ionic, osmotic, and oxidative stresses, and with heat shock, in addition to stress proteins related to protein folding/unfolding processes. It was proposed that the peptide segment on HAFP may represent a topographic 'hotspot', sensitive to stress/shock conditions, which exhibits a propensity for conformational alteration in the tertiary structure of the fetal protein. PMID- 12913350 TI - Evaluation of prolonged fetal monitoring with normal and pathologic outcome probabilities determined by artificial neural network. AB - OBJECTIVE: The purpose of this study was to objectively evaluate prolonged fetal heart rate (FHR) monitoring, which has been difficult to do with conventional cardiotocogram (CTG). METHODS: FHR was analyzed by an artificial neural network computer that calculates probabilities of normal, pathologic and suspicious outcome. Earlier normal and pathologic outcome probabilities (OPs) recorded during 15-min intervals are averaged every 5 min. Initially, two curves (the averaged normal and pathologic OPs) are compared. Furthermore, a single curve traced for each difference between the averaged normal and pathologic OPs and its value are studied. Our FHR probability data are of 9 cases reported in a previous paper on neural network FHR analysis. RESULTS: In the 4 cases of normal neonatal condition, the trends of the averaged curves and the last averaged values were higher for normal OP than for pathologic OP, and the final values of the difference were >0. On the other hand, in the 5 cases of neonatal depression, the trend of the two curves and the final values were lower for normal than for pathologic OP; and the final difference values of averaged probabilities were <0. For prolonged monitoring, the single parameter is more useful than the comparison of the two curves. CONCLUSION: A useful single parameter is obtained for the accurate and objective evaluation of prolonged FHR monitoring. The present method is promising for prospective studies using the combined system of experts and neural computers. PMID- 12913351 TI - Studies on the isolation and identification of fetal nucleated red blood cells in the circulation of pregnant women before and after chorion villus sampling. AB - OBJECTIVE: To investigate the feasibility of various molecular forms of hemoglobin as markers for fetal nucleated red blood cells (NRBCs). METHODS: The presence of epsilon and gamma globin positive NRBCs was investigated in pure fetal blood and in blood from pregnant women before and after chorion biopsy. Maternal samples were enriched for NRBCs by various conventional methods, including limited enrichment by only positive CD71 selection or single density gradient. We searched for fetal cells on slides by automated scanning. Fetal cells were defined by (1) the presence of epsilon or gamma globin and (2) simultaneously by the presence of a Y chromosome signal. RESULTS: 18 of 25 gamma globin positive cells identified in blood samples after chorion biopsy were chromosome Y signal positive, and 1 cell had two X chromosome signals. 263 of 339 epsilon globin positive cells identified in blood samples after chorion biopsy were hybridized with X and Y chromosome probes. None had two X signals, and 249 were Y positive. In blood samples before chorion biopsy, only 1 epsilon globin positive fetal NRBC and no epsilon globin positive maternal NRBCs were found. CONCLUSIONS: Epsilon globin may be specific for fetal NRBCs. Only 1 epsilon globin positive fetal cell was identified in 1 of 12 blood samples before chorion biopsy, representing a total of 182 ml of maternal blood. This suggests that most fetal cells found in maternal blood by fluorescence in situ hybridization methods may not be NRBCs. PMID- 12913352 TI - Percutaneous ultrasound-guided injection of the trachea in fetal sheep: a novel technique to target the fetal airways. AB - OBJECTIVE: To access the fetal airways percutaneously using ultrasound-guided injection of the fetal trachea in sheep. METHODS: Adenoviral gene therapy vectors and transduction-enhancing agents were delivered to the trachea via a needle inserted through the thorax or the neck of late-gestation (0.9 term, n = 3) or mid-gestation (0.5-0.8 term, n = 18) fetal sheep using ultrasound guidance. RESULTS: Injection of the trachea in the fetal thorax was successful in 16 out of 18 fetuses and achieved at the first attempt in 9 fetuses within 12 min [mean 7 min and 31 s +/- (SD) 3 min and 4 s]. Survival was 100%. Injecting the trachea in the neck was less successful. CONCLUSIONS: The fetal trachea of the sheep can be safely accessed by percutaneous ultrasound-guided injection to deliver vectors directly to the fetal airways for gene therapy. It may also enable tracheal occlusion for the antenatal treatment of congenital diaphragmatic hernia without the need for endoscopy or open surgery. PMID- 12913353 TI - Evidence for recombination in the envelope gene of maedi-visna virus. AB - Frequent recombination occurs during replication in all retroviruses examined. This increases the genetic variation in the retroviral population and may be of importance in the evolution of the virus. Maedi-visna virus (MVV), a retrovirus of sheep, has a highly variable envelope gene. In a previous experiment, 20 sheep were infected with an uncloned strain of MVV and virus was isolated at regular intervals for 7 years. We sequenced the envelope genes of a number of these strains and found evidence for recombination that may have contributed to the observed high frequency of antigenic variants. PMID- 12913354 TI - Sequence analysis of the 3' end of three Grapevine fleck virus-like viruses from grapevine. AB - The 3' end of the genome of three Grapevine fleck virus-like viruses, i.e. Grapevine redglobe virus (GRGV), Grapevine asteroid mosaic-associated virus (GAMaV), and an unidentified virus from a Greek grapevine (accession GR8-19) was amplified from reverse transcribed total nucleic acid extracts from infected grapevine tissues and sequenced. The analysed genome portions differed in size and organization. The 3' ends of GAMaV (1852 nt) and of GR8-19 (1791 nt) resembled that of marafiviruses, as both encoded a single putative polyprotein containing the conserved "marafibox" sequence and lacked the stop codon between the replicase and coat protein genes. By contrast, the replicase and coat protein genes present in the terminal 2006 nt of GRGV genome were clearly separated and there was a 3'-proximal open reading frame encoding a putative proline rich protein with molecular mass of c. 17 kDa. The genome of all three viruses was polyadenylated. The organization of the 3' terminal genomic region and phylogenetic analysis of viral replicases and coat proteins suggest that GAMaV and the Greek virus GR8-19 belong in the genus Marafivirus, and GRGV in the genus Maculavirus, family Tymoviridae. Virus GR8-19 had molecular traits differing enough from GAMaV and other marafiviruses to be regarded as a new putative species in the genus Marafivirus, for which the name of Grapevine rupestris vein feathering virus is proposed. PMID- 12913355 TI - Functional characterization of the ecdysteroid UDP-glucosyl transferase gene of Helicoverpa armigera single-enveloped nucleopolyhedrovirus isolated in South Africa. AB - The ecdysteroid UDP-glucosyltransferase (egt) gene of a single enveloped nucleopolyhedrovirus was located using an Hz-SNPV gene-specific probe. This SNPV was found infecting a colony of Helicoverpa armigera (HaSNPV) in the Western Cape region of South Africa. The open reading frame of the HaSNPV-SA egt is 1.548 nucleotides long and encodes a predicted protein of 516 amino acids with a Mr of 58,897-kDa. The 5'-noncoding region contained an early transcription initiation motif (CAGT) and a baculovirus late transcription motif (ATAAG). A transcription enhancer sequence (GATA) was also identified. Two possible TATA boxes together with an AT rich region were also recognized. A putative signal peptide of 20 residues was present at the N-terminus of the predicted EGT sequence. A polyadenylation signal (AATAAA) was found downstream of the translation stop codon. Five Helicoverpa NPV EGT's that have an extremely high degree of nucleotide and amino acid sequence homology were used in this study. Single nucleotide polymorphisms (SNPs) within the gene were tabulated. The Helicoverpa NPV egts seem to be closely related to the egt genes of Mamestra configurata NPV (MacoNPV), Buzura suppressaria NPV (BusuSNPV) and Spodoptera exigua NPV (SeMNPV) with amino acid identities of approximately 50%. The Helicoverpa NPV EGTs show ten conserved motifs with other EGTs. A phylogenetic tree of 27 baculovirus EGTs and a human UDP-glucoronosyltransferase was constructed using Neighbour-joining within CLUSTAL X. That a secreted and active EGT is encoded by HaSNPV-SA was confirmed by assay of infected cell culture medium. PMID- 12913357 TI - Coat protein sequence analysis reveals occurrence of new strains of Sweet potato feathery mottle virus in Uganda and Tanzania. AB - The 3'-proximal part (1.8 kb) of the Sweet potato feathery mottle virus (SPFMV) genome was studied in four SPFMV isolates collected from farmers' fields in western Uganda (SPFMV-Bny), eastern Uganda (SPFMV-Sor) and Bagamoyo district, Tanzania (SPFMV-TZ1 and SPFMV-TZ2). Unlike the other three SPFMV isolates, SPFMV Sor was not detected with the polyclonal antisera to SPFMV. It showed moderately high coat protein (CP) nucleotide (93.3-96.7%) and amino acid (93.6-96.8%) sequence identity to the isolates of the SPFMV strain group C. In contrast, identities (78.1-80.1%, and 79.9-83.1%) to isolates of the SPFMV strain groups O, RC, and the East African (EA) strain group were low. Similar to some isolates (SPFMV-CH2 and SPFMV-6) of strain group C, but different from other SPFMV isolates, SPFMV-Sor contained a deletion of 6 nucleotides in the CP-encoding region (CP amino acid positions 62-63). Phylogenetic analysis of the CP sequences indicated that SPFMV-Sor belongs to the SPFMV strain group C that has not been reported from Africa. Sequence data were obtained for the first time from Tanzanian SPFMV isolates in this study, and phylogenetic analysis indicated that they belong to the strain group EA, which is unique to East Africa. PMID- 12913356 TI - Molecular anatomy of Tupaia (tree shrew) adenovirus genome; evolution of viral genes and viral phylogeny. AB - Adenoviruses are globally spread and infect species in all five taxons of vertebrates. Outstanding attention is focused on adenoviruses because of their transformation potential, their possible usability as vectors in gene therapy and their applicability in studies dealing with, e.g. cell cycle control, DNA replication, transcription, splicing, virus-host interactions, apoptosis, and viral evolution. The accumulation of genetic data provides the basis for the increase of our knowledge about adenoviruses. The Tupaia adenovirus (TAV) infects members of the genus Tupaiidae that are frequently used as laboratory animals in behavior research dealing with questions about biological and molecular processes of stress in mammals, in neurobiological and physiological studies, and as model organisms for human hepatitis B and C virus infections. In the present study the TAV genome underwent an extensive analysis including determination of codon usage, CG depletion, gene content, gene arrangement, potential splice sites, and phylogeny. The TAV genome has a length of 33,501 bp with a G+C content of 49.96%. The genome termini show a strong CG depletion that could be due to methylation of these genome regions during the viral replication cycle. The analysis of the coding capacity of the complete TAV genome resulted in the identification of 109 open reading frames (ORFs), of which 38 were predicted to be real viral genes. TAV was classified within the genus Mastadenovirus characterized by typical gene content, arrangement, and homology values of 29 conserved ORFs. Phylogenetic trees show that TAV is part of a separate evolutionary lineage and no mastadenovirus species can be considered as the most related. In contrast to other mastadenoviruses a direct ancestor of TAV captured a DUT gene from its mammalian host, presumably controlling local dUTP levels during replication and enhance viral replication in non-dividing host tissues. Furthermore, TAV possesses a second DNA-binding protein gene, that is likely to play a role in the determination of the host range. In view of these data it is conceivable that TAV underwent evolutionary adaptations to its biological environment resulting in the formation of special genomic components that provided TAV with the ability to expand its host range during viral evolution. PMID- 12913358 TI - High-resolution methylation analysis and in vivo protein-DNA binding at the promoter of the viral oncogene LMP2A in B cell lines carrying latent Epstein-Barr virus genomes. AB - Latency protein LMP2A of Epstein-Barr virus (EBV) has been implicated in EBV related tumorigenesis. To understand the host cell dependent expression of the LMP2A gene, it is necessary to analyse the regulatory mechanisms of the LMP2A promoter (LMP2Ap). By transient transfection and in vitro binding analyses two CBF1 sites have previously been shown to be involved in the regulation of LMP2Ap. However, the promoter structure has not been examined at the nucleotide level in vivo. Therefore we undertook a comprehensive analysis of in vivo protein binding and of CpG-methylation patterns at LMP2Ap in a panel of B cell lines carrying latent EBV genomes. The presence of characteristic footprints on two CBF1 and further binding-sites, together with overall hypomethylation of CpG dinucleotides correlated well with promoter activity. In contrast, the absence of several genomic footprints, as well as the presence of patches of highly methylated CpG dinucleotides were characteristic of silent LMP2Aps. PMID- 12913359 TI - Characterization of RNA-dependent RNA polymerase activity of CSFV NS5B proteins expressed in Escherichia coli. AB - The full-length NS5B protein, and the truncated NS5B proteins of classical swine fever virus (CSFV) resulted from deletion of 24, 36, 65 or 82, amino acid residues at the C terminal were expressed in Escherichia coli cells and purified with a C-terminal hexahistidine tag. In addition to the full-length NS5B protein, those truncated NS5B proteins with deletion of 24, 36, or 65 amino acid residues were demonstrated to have RNA-dependent RNA polymerase (RdRp) activity, which was not found in the truncated NS5B proteins with deletion of 82 amino acid residues. Analysis of the template specificity of CSFV RdRp was done containing the different NS5B proteins with RdRp activity. It was shown that the template specificity of the enzyme was not strict with NS5B proteins truncated, suggesting that the C terminal of CSFV NS5B protein was involved in the template specificity of the enzyme. Site-directed mutagenesis of and prediction of the secondary structure of 3' terminal sequence of the template indicated that the cytidines at 3' terminus and the correct secondary structure of the template were essential to initiation of RNA synthesis by RdRp. Oxidation of the hydroxyl groups of the RNA template revealed that both the de novo initiation mechanism and the template priming mechanism preference might be employed by the CSFV RdRp. PMID- 12913360 TI - Expression of VP2 gene protein of infectious bursal disease virus detected in Korea. AB - The VP2 gene DNA (1.4 kb in approximate) of a very virulent infectious bursal disease virus (vvIBDV) Chinju strain detected in Chinju, Korea was cloned into the bacmid, a baculovirus shuttle vector, through transposition of the gene from initially cloned pFastBacHTa plasmid, a baculovirus expression vector, and was subsequently expressed in Spodoptera frugiperda (Sf) cells. Biological properties of the expressed VP2 subunit protein were characterized to aid in the development of genetically engineered diagnostic reagents and vaccines against the vvIVDV. When the VP2 DNA-recombinant bacmid was transfected and propagated in the Sf cells, the cells showed no occlusion formation, which is a positive evidence for the insertion of the VP2 DNA into the polyhedrin gene of the bacmid, whereas the occlusions were observed in the cells infected by the Autographa californica nuclear polyhedrosis virus, a wild baculovirus. The expression of VP2 DNA was identified by strong positive reaction in fluorescent antibody test using chicken anti-IBDV serum. The VP2 protein was determined as a polypeptide band with Mr of 48 kDa by the sodium dodecyl-polyacrylamide gel electrophoresis for the lysate of the Sf cells infected with the recombinant bacmid. The VP2 protein was successfully purified from the cell lysate by Ni-NTA affinity chromatography. The expressed VP2 subunit protein reacted specifically with chicken anti-IBDV serum in Western blotting. PMID- 12913361 TI - Expression of the genomic form of the bovine viral diarrhea virus E2 ORF in a bovine herpesvirus-1 vector. AB - Bovine viral diarrhea virus (BVDV) is a ubiquitous pathogen of cattle with a world-wide distribution. Recently, the possibility of using recombinant virus vectors to immunize cattle against selected BVDV genes has gained widespread interest. Among the virus vectors tested, bovine herpesvirus-1 (BHV1) provides many unique advantages. However, results of recent studies have raised the possibility that the codon usage pattern required for optimal expression in a BHV1-infected cell may be incompatible with the codon usage pattern of BVDV. If true, use of BHV1 to express BVDV proteins would require construction of synthetic BVDV genes that have been modified to resemble the codon pattern of BHV1. To explore this possibility, we constructed a BHV1 recombinant containing the genomic form of the BVDV (NADL) E2 ORF and compared expression of the E2 protein with that of the endogenous BHV1 gD protein. We observed that E2 was expressed at a significant rate compared to that of the gD protein. We conclude that codon usage problems are unlikely to constitute a serious problem for expression of BVDV proteins in BHV1 vectors. PMID- 12913362 TI - Molecular characterization of the major virion protein gene from the Trichoplusia ni ascovirus. AB - Ascoviruses (AVs) belong to a family of double-stranded DNA viruses that infect Lepidoptera insects and cause the unique pathology of forming virion-containing vesicles in the hemolymph of infected hosts. Virions of AVs are large and contain more than 12 polypeptides. A gene, TnAV-CP, encoding the major structural protein of the Trichoplusia ni ascovirus 2a (TnAV-2a) was cloned by immunoscreening an expression library with antibodies against total TnAV virion proteins. TnAV-CP is an intronless gene with an open reading frame encoding a protein of 455 amino acids. Southern blot showed that it is a single copy gene. A 3.8 kb BamHI fragment containing the complete TnAV-CP gene was cloned and sequenced. Northern analysis detected the transcription of the 1.4 kb TnAV-CP mRNA from 24 h after infection. The predicted TnAV-CP protein was expressed in bacterial expression system and purified to homogeneity. The recombinant protein was used to affinity purify specific antibodies from the antiserum. The purified antibodies reacted strongly with a single protein of approximately 52 kDa from the total TnAV virion proteins in a Western blot. This protein corresponds to the most abundant structural protein present in the virions of several AVs. Sequence comparison showed that TnAV-CP is most homologous to the putative capsid proteins from AVs infecting noctuid insects, less homologous to that of Diadromus pulchellus ascovirus 4a (DpAV-4a), further supporting the distinction of two subgroups within the family Ascoviridae. Phylogenetic analysis using the putative capsid protein suggested that AVs were closely related to members of Iridoviridae, which corroborated the result based on DNA polymerase delta sequences. The apparent differences between Ascoviridae and Iridoviridae in host range, virion morphology, and genome configuration, and the similarities in genes and methylation of genomic DNA were discussed. PMID- 12913363 TI - Evolutionary history of Hepatitis B virus genotype F: an in-depth analysis of Argentine isolates. AB - A revised analysis on the evolutionary history of hepatitis B virus (HBV) genotype F is herein presented with the incorporation of two new complete genomes from Argentina. The study of the phylogenetic-tree topology, genetic distances, and amino acid mutations confirmed with high reliability the existence of four different genetic clusters of this genotype. Argentine isolates were located in two groups of viruses that showed a great inner homogeneity but, interestingly, divergence between them was in the order of that existing among groups from different locations. Although the origin of these two viral populations is not clear, they do not seem to derive from each other, therefore the existence of at least two founder viral populations in Argentina is a more acceptable explanation. PMID- 12913365 TI - Learning the language of abstinence in addiction treatment: some similarities between relapse-prevention with disulfiram, naltrexone, and other pharmacological antagonists and intensive "immersion" methods of foreign language teaching. AB - Relapse-prevention (RP) is an educational process. Learning to abstain from alcohol or opiates after years of dependence involves selectively suppressing old, maladaptive habits of thought and behavior and establishing new, adaptive ones. This process resembles foreign language (FL) learning. Effective FL teaching techniques are relevant to RP. "Immersion," the most effective FL teaching method, discourages students from using their first language ab initio, requiring them to use the FL instead, however inexpertly. It resembles exposure and response-prevention for phobic or compulsive disorders. Supervised disulfiram aids RP by discouraging alcoholics from responding to real-life drinking cues in the "language" of excessive drinking, requiring them, ab initio, to practice new, alcohol-free responses. Supervised or depot naltrexone acts similarly in opiate dependence. We discuss the concept of antagonist-assisted abstinence. PMID- 12913366 TI - Evaluation of medical student attitudes toward alcoholics anonymous. AB - This is a two-phase study on attitudes of medical students toward Alcoholics Anonymous. The first phase compares views of addiction faculty to third-year medical students on the importance of spirituality in addiction treatment. We administered a questionnaire to assess attitudes toward spiritual, biological, and psychosocial approaches to addiction treatment. The faculty viewed spirituality as relatively more important in addiction treatment than did the students. The second phase was designed to assess whether medical student attitudes toward spiritually based treatments changed over the course of a psychiatry clerkship. At the beginning of the clerkship, students rated a spiritually oriented approach as important in addiction treatment as a biological approach, whereas, at the end of the clerkship, they rated the biological approach as more important. It may be important to educate medical students about the spiritual dimensions of recovery so they can integrate this into their treatment of addiction. PMID- 12913364 TI - Medical specialization, profession, and mediating beliefs that predict stated likelihood of alcohol screening and brief intervention: targeting educational interventions. AB - Practitioner-level educational approaches that promote screening and brief intervention (SBI) seldom consider providers' profession and medical specialization. Strategies that consider these variables may be better equipped to affect change in beliefs and behavior. The aim of this study was to identify beliefs that predict stated likelihood of practicing SBI by specialty and health profession in order to guide the direction of educational strategies. Physicians and nurse practitioners were studied that specialized in family, internal, obstetric gynecology (ObGyn), and pediatric medicine. The results indicated that independent of amount of previous postgraduate alcohol education and knowledge, self-rated competence mediated between specialty and likelihood of practicing SBI. For instance, low self-rated competence for ObGyn was a barrier that suppressed likelihood of practicing SBI. Other findings were that role legitimacy mediated the association between profession and likelihood of SBI, so that lack of role legitimacy was a barrier for physicians but not for nurse practitioners. We suggest that targeted educational strategies for ObGyn and pediatric clinicians may prove more effective than the prevalent one-size-fits all approaches aimed at general adult populations. PMID- 12913367 TI - A case of factitious disorder presenting as alcohol dependence. AB - Factitious disorders involve the feigning of physical or psychological symptoms in order to assume the patient role. Pseudologia fantastica, which involves the pathological creation of fabrications about one's background, is an associated feature of factitious disorders. Substance abuse disorders are also associated with factitious disorders. However, the manifestation of factitious complaints as substance abuse has yet to be reported in the literature. This case study describes a patient referred to a residential substance abuse treatment program who was discovered to have factitious alcohol dependence and prominent pseudologia fantastica. PMID- 12913368 TI - Perceptions of smoking among African American light smokers. AB - Despite smoking fewer cigarettes per day, African Americans extract more nicotine per cigaretts smoked, and have higher than expected tobacco-related morbidity and mortality. The present study explores perceptions about smoking and motivations for smoking cessation among African American light smokers. Four focus groups consisting of 22 adults were conducted at an inner-city community health center. Participants were recruited through flyers and referral by clinic staff. Two major themes were identified: (1) similar smoking patterns, and (2) perceived differences from heavy smokers. Many participants perceived themselves as addicted to nicotine, experienced withdrawal symptoms with past quit attempts, and are interested in smoking cessation. Perceptions about nicotine addiction, difficulty with quitting, and interest in smoking cessation suggest a need to design and test cessation interventions among this understudied population. PMID- 12913369 TI - Primary care residents self assessment skills in dementia. AB - The ability to accurately self-assess is a critical component of professionalism and is included in the newly required Accreditation Council of Graduate Medical Education (ACGME) core competencies. To assess residents' ability to accurately self-assess their competencies related to a commonly presenting problem in geriatrics, a Standardized Patient, portraying an individual with early signs of dementia, was inserted into family medicine residents' clinic schedules. Immediately post the encounter, each resident self-assessed his/her performance using a four category (Communication, History of Present Illness, Social History, Functional Assessment), 17-item behavioral checklist. The items in each category highlighted items specific to a dementia-screening interview (e.g., HPI: Used a standardized exam which includes orientation, memory, recall and registration). Resident ratings were compared to ratings from two faculty assessors who independently viewed the videotape of each resident's SP interview. While statistically significant differences between the self-assessment and expert assessors appeared in only one of the four major checklist categories (functional assessment), item specific analysis revealed significant differences on discrete items within the dementia screening interview. Implications for teaching and assessment consistent with the ACGME required competency assessment category of professionalism are discussed. PMID- 12913370 TI - Core characteristics of the competent general practice trainer, a Delphi study. AB - INTRODUCTION: The specific skills, attitude, knowledge, and personality characteristics, which should define the competent GP-trainer have been subject of research for many years. What are the most important of these characteristics have yet to be delineated. AIM: The aim of this study is to identify which characteristics are prerequisite for a competent GP-trainer. METHOD: A modified Delphi procedure was used to answer the research question. Earlier qualitative research using focus groups identified 43 characteristics defining the competent GP-trainer. These characteristics were the subject of the Delphi procedure in which 48 experts chosen from the entire field of Dutch General Practice Training participated. RESULTS: After two rounds of the Delphi procedure, 37 characteristics were identified as being important for a competent GP-trainer. The characteristics which were ranked as the most important were 1. a competent GP-trainer is good at giving feedback, 2. dares to give feedback, 3. is critical of the GP-trainee and the learning process, 4. is good at communicating with the GP-trainee, and 5. has respect for the trainee. An important new characteristic has been identified: "being able to inspire reflection in the trainee". CONCLUSION: The core characteristics of the GP-trainer as identified in this research project, form a profile for the ideal GP-trainer to be used as a blueprint for objectives of schooling and for the assessment of GP-trainers. PMID- 12913371 TI - The effect of longterm vs shortterm tutors on the quality of the tutorial process and student performance. AB - Small-group, problem- based learning can require a significant amount of individual faculty time when groups last as long as 10 weeks. One solution is to use two short-term (3-5 weeks) tutors instead of a single long-term (6-10 weeks) tutor. This study was performed to evaluate whether having short-term instead of a long-term tutors affected student performance and/or the quality of the tutorial process. The grade point averages for first and second year medical students in the classes of 2001 and 2002 were stratified by how many long-term tutors students had (2, 3, 4, 5, or 6) over a period of 17 months. In addition, students completed anonymously a 24-item tutor evaluation questionnaire utilizing a 5 point Likert scale (outstanding, good, satisfactory, marginal or unsatisfactory) at the mid-point and at the end of each tutorial. The evaluations were stratified by whether the tutor was short-term or long-term (to the mid point or end of the tutorial, respectively). Statistical analysis was performed using analysis of variance. Grade point averages did not correlate with the number of long-term tutors that students had. Long-term tutors were rated significantly higher than short-term tutors in 17 of the 24 categories evaluated by the questionnaire, including development of students' critical thinking, information skills and self-learning skills; encouraging students to express themselves freely and to take responsibility for their own learning; helping students to balance basic science and clinical applications; and overall assessment. Short-term vs long-term tutors do not affect objective student performance as measured by grade point average. However, long-term tutors are ranked significantly higher by students in numerous areas related to the perceived quality of the tutorial process. Short-term tutoring may be less of a demand on an individual faculty's time, but may lessen the students' perceived quality of the tutorial process. PMID- 12913372 TI - Monitoring medical students' professional attributes: development of an instrument and process. AB - PURPOSE: To describe and evaluate the implementation of an instrument and process to assess and monitor professional attributes in medical students. METHOD: The instrument was developed following a survey of professional attributes of concern displayed by medical students. This was followed by development and validation, amongst medical faculty staff and students, of the instrument and its supporting process. Evaluation was by recording participation rates by staff and by determining the consistency of the instrument across dimensions and across clerkships. Implementation in practice is described by using a case study based on one school's first year of use amongst all the students. RESULTS: In its first year of use 7,418 forms were distributed to 376 students and 6,621 forms were returned (89%). The instrument's reliability was greatest where staff were diligent in ensuring thorough implementation. The process was able to detect students of concern and provide effective remediation and ongoing monitoring. CONCLUSION: The process that has been developed is sensitive, robust and is able to detect, monitor and remedy attributes traditionally regarded as difficult to define and defend. The strengths of the process lie in (1) clear definitions of acceptable and unacceptable behaviors, (2) reliance on observations by more than one person and over more than a single time period, (3) the ability to provide a longitudinal view of student's progress. PMID- 12913373 TI - A qualitative study of the attitudes to teamwork of graduates from a traditional and an integrated undergraduate medical course. AB - INTRODUCTION: In 1994 the University of Manchester medical school introduced an integrated undergraduate course using problem-based learning throughout. This study explores differences between the new and old (traditional) course graduates' attitudes to, and conceptualization of, teamwork. METHODS: Semi structured interviews were conducted with 24 graduates of the traditional course (graduating in 1998) and 23 from the new course (1999 graduates), representing approximately 14% of graduates from each cohort. Theories were then developed from concepts emerging from the data. RESULTS: The new course graduates (NCGs) had a broader view of members of a health professional team. The NCGs believed that the medical team should provide support and were more comfortable consulting them when faced with problems. CONCLUSIONS: The new curriculum has had some impact on conceptualization and attitude to teamwork. However, further development is required if graduates are to see themselves as part of a multi professional team. PMID- 12913374 TI - Are family practice residents able to interpret electrocardiograms? AB - BACKGROUND: Teaching electrocardiogram (ECG) interpretation is a recommended component of the family practice residency curriculum. Published information concerning the ECG interpretation ability of residents is sparse. This study sought to ascertain the baseline knowledge of family practice residents' ECG interpretation skills and extent of improvement after one year of training. METHODS: A 15 ECG examination was administered to 38 PG-1 and 14 upper level residents at 5 residency programs at the beginning of the academic year and to residents at the authors' program at the end of the academic year. Pre-test scores among the five programs were compared using an analysis of variance (ANOVA). Pre-test and post-test scores were compared using a paired randomization test. RESULTS: No difference was found between average scores from each site, or between the beginning and end of the academic year. Residents were more likely to misinterpret items such as myocardial infarction, myocardial ischemia, and atrial fibrillation. CONCLUSIONS: Residents in family practice have considerable deficiencies in ECG interpretation skills. Further studies are needed to determine effective ECG teaching curricula. PMID- 12913375 TI - Context-dependent memory in a meaningful environment for medical education: in the classroom and at the bedside. AB - PURPOSE: Learning-in-context is a much-discussed topic in medical education. Information is said to be better recalled when the learning environment resembles the later retrieval environment. Godden and Baddeley (1975) showed that divers recalled words better when the recall condition matched the original learning environment, i.e. underwater or on land. Though it is unclear whether the findings can be generalized for medical education, medical educators regularly refer to them. We replicated the Godden and Baddeley study in ecologically more valid conditions for medical education and extended it with meaningful subject matter (namely, a patient case description). METHOD: Sixty-three clerks were randomized over four conditions, contrasting a clinical (bedside) with an educational (classroom) environment as both learning and recall conditions. Students were asked to recall a list of words and a patient case in the same environment or in the opposite environment as where they learned it. RESULTS: We failed to find a significant same-context advantage for free recall of the list of words and the patient case propositions. However, there does appear to be a slight tendency towards better recall of the case description when learning took place in the clinical environment. DISCUSSION: In medical education, the context, if conceived as physical surroundings, does not seem to contribute to a same context advantage. One should be cautious in generalizing the findings of Godden and Baddeley. However, different forms of 'context' other than the physical one used in the Godden and Baddeley study may well enhance learning effects in medical education. PMID- 12913376 TI - Advice for young investigators: historical perspectives on scientific research. AB - Physicians attempting to carry out well-executed scientific investigations require an historical perspective; an awareness of our ignorance; an imagination unencumbered by scientific prejudices; originality, curiosity and imagination; patience and persistence in the face of frustration; adaptability and perseverance; a critical spirit of evaluation and willingness to question dogma; devotion to the search for truth; and an awareness of those responsibilities to individuals and to society which define the medical profession. This paper explores, through the words of famous scientists, philosophers and authors, the process of scientific investigation and discusses those qualities that investigators can develop to become more insightful and successful researchers. PMID- 12913377 TI - The ethics of gambling research: an agenda for mature analysis. AB - The purpose of this article is to explore the ethics of both the anti-gambling and pro-gambling groups as they present their research into the gambling phenomenon. The type of ethical thinking that each group uses will be characterized to show how their ethical views have caused their research to be biased. The second part of this article proposes a modest research agenda for future gambling studies. This agenda rests a foundation of a "mature" ethical thinking. PMID- 12913379 TI - Pursuing "An agenda for mature analysis". PMID- 12913378 TI - Method, method: who's got the method? What can we know about the number of compulsive gamblers? PMID- 12913380 TI - The courage to be counted. PMID- 12913381 TI - Recommendations for research and public policy in gambling studies. AB - The purpose of this article is to respond to and expand on the ideas presented by Father McGowan in The Ethics of Gambling Research: An Agenda for Mature Analysis. We provide specific recommendations for future research and public policy in the field of gambling studies. We suggest that key conceptual definitions--such as, problem, compulsive, and pathological gambling--should be clarified, established, and distinguished from one another before gambling research is conducted. Proper methodological procedures are recommended, where power analyses, pilot studies, and representative samples are appropriately conducted and analyzed. Retrospective and Prospective studies are considered and differentiated while Discrete Time Event History Analysis--namely, Life Tables Analysis and Discrete Time Logistic Regression--are proposed. PMID- 12913382 TI - Building a National Lottery: reviewing British experience. AB - The launch of the United Kingdom's National Lottery at the end of 1994, was one of the most significant developments in both the funding of Britain's cultural sector and the British gambling sector since 1946. Thus in the 1995 financial year National Lottery ticket sales exceeded pound 5 billion of which 28% went to five specified "good causes." This paper presents an overview of the National Lottery's launch and growth to date using published material and data. The lottery's establishment and regulatory regime are discussed, together with its impact in terms of both economic activity and the wider betting and gaming market within the UK. The National Lottery is then considered within the broader international context, and the incoming Labour Government's restructuring plans aimed at creating a "People's Lottery" are discussed. Thus the paper provides an overview of the development of a major new national institution. PMID- 12913383 TI - The correction for bias in prevalence estimation with screening tests. AB - The concern that the South Oaks Gambling Screen (SOGS) and other screening tests have a relatively high rate of false positive errors which results in overestimation of the true prevalence in general population studies is shown to be unfounded. False positives are seen to be a necessary but not sufficient condition for overestimation. It is demonstrated that the proper research question is whether the sample prevalence estimator is biased, and, if so, in which direction. One solution to the problem of bias is shown to depend on the availability of estimates of the error rates of the test. PMID- 12913384 TI - College student perceptions of crime and casino gambling: a preliminary investigation. AB - To gain insight into how attitudes toward casino gambling vary by an area's experience with gambling, a survey was given to 415 university students at the University of Nevada, Reno, and 420 students at the University of Memphis. Casino gambling has been legal in Reno since 1931, but was introduced in the Memphis area in 1992. Given the differences in duration and degree of integration of casinos in each area, significant differences were expected by area in respondents' perceptions of casinos and casino gambling. While some significant differences were found, the most prominent finding is the degree of similarity of attitudes of students in each university and the high percentage in each sample that associated casino gambling with types of crime. Where significant differences did exist, the findings are interpreted as supporting either an exposure hypothesis or a cultural conservatism hypothesis. PMID- 12913385 TI - Familial and social influences on juvenile gambling behavior. AB - Social learning theory maintains that individuals model, learn and maintain behaviors that are observed, appealing and reinforcing. As such, parents and family members can often serve as significant models for gambling. Four hundred and seventy seven children between the ages of 9 and 14 completed a questionnaire inquiring about their gambling activities, including where and with whom gambling occurs, as well as information concerning their perceptions of their own gambling behavior. Results indicate that 86% of children who gamble regularly reported gambling with family members. Fifty-three percent of students who gambled within the previous 12 months reported gambling with their siblings, 40% gambled with their parents, 46% gambled with other relatives, and 75% gambled in their own homes. Students' responses also indicated gambling with their friends (75%), gambling alone (18%), and with strangers (8%). As children's age increases they tend to gamble more at friend's homes and at school. Prevalence rates indicated that 81% of the total sample had gambled at one point in their lives and 52% of those children reported gambling once a week or more. Eleven percent reported that gambling makes them feel important, 27% feel they gamble more than they desire to do so, and only 10% of the grade 8 students fear being caught gambling, suggesting gambling activities to be a socially acceptable behavior. Several clinical and research questions are addressed. PMID- 12913386 TI - DSM-IV and the South Oaks Gambling Screen: diagnosing and assessing pathological gambling in Turkey. AB - The main purpose of this study was to investigate the effectiveness of the DSM-IV diagnostic criteria and the South Oaks Gambling Screen (SOGS) in identifying Turkish pathological gamblers. Fifty-nine subjects participated in the study. The subjects were diagnosed as either pathological gamblers or not (comparison group) through the use of the DSM-IV criteria and were given the Turkish version of the SOGS. Four of the ten DSM-IV criteria were found to be problematic in the diagnosis of Turkish pathological gamblers. The data concerning reliability and validity of the Turkish version of the SOGS suggested that the SOGS can be used as a reliable and valid instrument in identifying Turkish pathological gamblers. Most (16 out of 20) of the items of the SOGS appear to work well in discriminating pathological gamblers from the subjects in the comparison group. In the case of the two DSM-IV criteria and the four SOGS items that failed to discriminate, cultural factors seemed to be responsible for the failure. PMID- 12913387 TI - Gambling activities of young Australians: developing a model of behaviour. AB - As more gambling venues open in Australia, youth gambling as problem behaviour has been identified. The aim of this study was to assess youth gambling in a population with easy access to gambling, and to evaluate the adequacy of a model for predicting adolescent gambling frequency and problem gambling. The model comprised a combination of the Theory of Reasoned Action (TRA) (Ajzen & Fishbein, 1980), personality variables (venturesomeness, impulsiveness), and cognitive bias variables derived from Weinstein's (1980) propositions concerning unrealistic optimism about future life events. A sample of 1017 school- and university-based adolescents indicated relatively low frequencies of gambling and low scores on the problem gambling scale, with males scoring higher than females on both measures. The TRA was supported with about 30% of the variance of each of gambling behaviour and problem gambling accounted for by intentions, attitudes, and subjective norms. Personality factors added significantly to the prediction of gambling. The cognitive bias variables, although independently not statistically significant, further contributed to prediction. PMID- 12913388 TI - "How much do you spend gambling?" ambiguities in survey questionnaire items. AB - The validity of a survey's findings is dependent upon the clarity and lack of ambiguity contained in each individual item yet minimal attention has been directed to this issue in most prevalence studies. Researchers have shown a tendency to accept the uncritical assumption that respondents interpret seemingly simple and straightforward items such as 'how much do you spend gambling?' in a consistent manner. No attempt is made to confirm the uniformity of responses by clarifying the mathematical formulae used by respondents to derive their answers. The purpose of this paper was to examine the consistency shown by a sample of 181 medical undergraduate subjects in estimating the level of gambling expenditure in a series of five case vignettes describing various scenarios of wins and losses during a session of gambling. Results revealed a wide variation in calculated figures depending upon whether or not subjects interpreted the item to mean net expenditure or turnover. Only two thirds or less of subjects calculated the figure to be the difference between the initial amount risked and the residual at the conclusion of the session. It is suggested that more attention be paid in prevalence and clinical studies to providing subjects with clear instructions on how to calculate expenditure estimates. PMID- 12913389 TI - Cognitive distortions in heavy gambling. AB - A sample of 38 regular and heavy gamblers, recruited through advertisements and not seeking treatment, were asked to describe special strategies, techniques or rituals that they used to increase their chances of winning at gambling in an open-ended interview. The mean South Oaks Gambling Screen Score for the sample was 7.7 with 64% of the sample scoring higher than 4. Their responses reflected multiple means by which the individual believed they were able to control (i.e., active illusory control, passive illusory control), reframe (i.e., interpretive control), or predict (i.e., probability control, predictive control) gambling outcomes. A larger number of cognitive distortions was associated with playing games in which skill was potentially a component (e.g., cards, sports) than in non-skill games (e.g., lotteries) as well as a positive family history of gambling. There were no sex differences. Implications of these findings for the cognitive psychopathology of gambling are discussed. PMID- 12913390 TI - A self-psychological study of a shared gambling fantasy in Eugene O'Neill's Hughie. AB - This paper presents an applied psychoanalytic study of Eugene O'Neill's two character play, Hughie. Applying the constructs of self psychology, the play illustrates both the narcissistic features and the emotional and behavioral characteristics of compulsive gamblers. The study focuses particular attention on the role of narcissistic fantasies--with both grandiose and megalomaniacal features--in affecting, temporarily, the mood of the characters. Moreover, it is shown that a shared gambling fantasy--a "winner among winners"--enables them to experience a sense of camaraderie, humanness, and the illusion of kinship. PMID- 12913391 TI - Gambling market and individual patterns of gambling in Germany. AB - In this paper individual patterns of gambling in Germany are estimated for the first time. The probit technique is used to test the influence of a set of individual characteristics on the probability of participating in each of the various legal games. A sample size of 1,586 adults collected for the pool of German lotteries provides a reliable set of data. All disaggregated estimations of participation are statistically significant at least at the 5 percent level. The basic findings suggest that gambling is a widespread normal (superior) consumption good because gambling participation tends to rise with income. Moreover, no demand anomaly can be found to justify assessing gambling as a social demerit. Only the participation in gaming machines is higher for younger, unemployed and less educated adults. While a moral evaluation of gambling is beyond the scope of this paper, the legislator's preference for a highly taxed state monopoly in gambling markets is to be rejected, at least for Germany. Additional statistical findings suggest distinct consumer perceptions of the characteristics of the various games and may be used for market segmentation. The paper starts with a descriptive introduction to the German gambling market. PMID- 12913392 TI - A preliminary consideration of cue-exposure, response-prevention treatment for pathological gambling behaviour: two case studies. AB - Cue-exposure, response-prevention, a promising treatment for cigarette-smoking, substance abuse, obsessive-compulsive disorder, etc. has not been fully utilized as a treatment for pathological gambling. Two case-studies, modelled on work by R. Self (1989), with cigarette smoking behaviour, report on an active and thorough application of this treatment to pathological gambling. In both cases, self-report of gambling behaviour and urges to gamble in any situation were found to decrease substantially. Outcome differences between participants were discussed with reference to different levels of pathological gambling as measured by the South Oaks Gambling Screen. It was concluded that cue-exposure, response prevention should be given more serious consideration as a treatment approach for pathological gambling. PMID- 12913393 TI - An empirical study of the impact of complexity on participation in horserace betting. AB - The aim of the research reported in this paper was to explore empirically whether levels of participation in horserace betting are affected by the complexity of the betting task. The study employed a systematic random sample of 1161 betting decisions made in UK offcourse betting offices during 1987. The research was conducted in a naturalistic setting where it was possible to grade complexity and to measure levels of participation. Complexity was defined in terms of both the number of alternatives in the decision-maker's choice set (number of horses in a race) and the complexity of the attributes set for each horserace (handicap vs. non-handicap races). Results indicated that bettors are not inhibited by alternative-based complexity, but may be inhibited to some extent by attribute defined complexity. PMID- 12913394 TI - Problem gambling in a federal prison population: results from the South Oaks Gambling Screen. AB - Three hundred and sixty three medium security federal prison inmates were interviewed with the South Oaks Gambling Screen (SOGS) with 5.2% of the sample achieving SOGS scores greater than 4 and another 7.4% attaining scores of 3 or 4, traditional markers of pathological gambling and problem gambling, respectively. Attempts were made to evaluate the relative accuracy of the continuum and dichotomy models of problem gambling by contrasting three groups of subjects (SOGS<3, SOGS=3 or 4, SOGS>4) on a series of gambling-related measures and comparing the relative ability of the SOGS and a dichotomized version of the SOGS to account for residual variance in these same gambling-related measures. Results were consistent with the view that problem gambling embodies features of both a continuum and dichotomy. Nearly half the subjects had desisted from problem gambling on their own, although self-remitters displayed less severe and extensive patterns of problem gambling than non-remitting subjects. PMID- 12913395 TI - Prevalence of gambling among Minnesota public school students in 1992 and 1995. AB - The purposes of this study were to examine the prevalence of gambling among youth, compare rates of gambling between 1992 and 1995, and determine what levels of gambling frequency may be considered common and uncommon. The two samples included 122,700 Minnesota public school students in the 6th, 9th, and 12th grades in 1992; and 75,900 9th and 12th grade students in 1995. Students were administered the Minnesota Student Survey, a 126-item, anonymous, self administered, paper-and-pencil questionnaire that inquires about multiple content domains, including gambling behaviors. The same questionnaire, with minor revisions to the gambling items, was administered in both 1992 and 1995 to students in their classrooms by the Minnesota Department of Education. There were slight decreases in overall gambling rates from 1992 to 1995. The majority of students gambled at least once during the past year. However, most did not play any game on a weekly/daily rate and did not report any problems associated with their gambling. Gender, grade, and race effects were found for gambling frequency. Boys gambled more often than girls, and 9th and 12th grade students gambled more often than 6th grade students. Asian American and White students reported lower rates of gambling frequency than Mexican/Latin American, African American, and American Indian students. From a statistical standpoint (i.e., beyond the 97.7 percentile), it may be considered in the uncommon range for girls to play two or more games at a weekly/daily rate, and for boys to play four or more games at a weekly/daily rate. Variables associated with gambling frequency included antisocial behavior, gender, and alcohol use frequency. Although the finding that gambling did not increase from 1992 to 1995 is encouraging, this is the first generation of youth to be exposed to widespread accessibility to gambling venues and gambling advertising and it will be important to continue monitoring the prevalence of youth gambling. PMID- 12913396 TI - Nevada gaming: revenues and taxes (1945-95). AB - In 1931, Nevada legalized casino gambling, an act which allowed its "gaming industry" to develop. Because many jurisdictions outside Nevada are now embracing legalized gambling as a path to a brighter economic future and because this proliferation presents Nevada with new challenges and opportunities, it is a good time to review the Nevada experience. Here, the focus is on gaming revenues and gaming tax collections during the 1945-95 period. We find that the growth rate of Nevada's gaming industry has slowed over time, with the relative importance of gaming in the Nevada economy peaking in the late 1970's. The Nevada economy has since been gradually diversifying, something which will increasingly pressure Nevadans to look for government revenue sources other than gaming if current levels of government services are to be maintained. PMID- 12913397 TI - Psychological characteristics of volunteers in studies on gambling. AB - Advertisements were published in local newspapers asking for volunteers to participate in a study on gambling. A battery of eleven questionnaires was mailed to the subjects assessing pathological gambling behavior, sociodemographic characteristics, motivation to gamble, erroneous perceptions about gambling, superstitious beliefs, depressive symptoms, social anxiety and avoidance, alcohol and drug abuse, problem-solving skills, and marital satisfaction. Subjects received $10 when they returned the questionnaires fully completed. Response rate was over 95%. Surprisingly, 29% of the respondents met the criterion for probable pathological gambling (score of 5 or more on the SOGS) and a further 16% were identified as potential pathological gamblers (scores of 3 or 4). The potential and probable pathological gamblers showed significant differences on motivational and cognitive variables related to gambling compared to those subjects who showed no signs of pathological gambling. The probable pathological gamblers reported significantly more signs of poor psychosocial functioning than the other two groups, including depressive symptoms, poor problem orientation, drug and alcohol abuse, and interpersonal conflict. The practical and theoretical implications of these results are discussed. PMID- 12913398 TI - Pharmacologic reperfusion therapy for acute myocardial infarction. AB - Acute myocardial infarction (MI) remains a significant problem in terms of morbidity, mortality and healthcare costs. Pharmacologic reperfusion therapies for MI are becoming increasingly complex. This review therefore places contemporary pharmacologic MI developments into perspective. An historical overview of pharmacologic reperfusion therapy for MI is provided, followed by an analysis of current limitations, treatment options, and present and likely future pharmacologic therapies. Adjunctive percutaneous and other treatments are also discussed, to clarify what is becoming a rapidly changing field. PMID- 12913399 TI - Effects of different thrombolytic treatment regimen with abciximab and tirofiban on platelet aggregation and platelet-leukocyte interactions: a subgroup analysis from the GUSTO V and FASTER trials. AB - BACKGROUND: Due to considerably high rates of reocclusion under standard thrombolytic therapy GP IIb/IIIa inhibitors have been combined with thrombolytics to improve therapeutic outcomes. Potential reasons for arterial reocclusion may be increased platelet activation, interaction of platelets with other cell types such as leukocytes and inadequate drug dosing due to lack of ideal platelet monitoring. We compared combination therapy regimens consisting of GP IIb/IIIa inhibitors and thrombolytics with respect to platelet inhibition and platelet leukocyte interactions. METHODS AND RESULTS: From the GUSTO V trial (standard rPA vs. reduced dose rPA and abciximab) and the FASTER trial (standard TNK-tPA vs. reduced dose TNK-tPA and tirofiban) 15 patients were monitored by platelet aggregometry, rapid platelet function assay (RPFA) and flow cytometry (FC). rPA alone (n = 5) caused initial increases in platelet aggregation. However, platelet aggregation was significantly (p < 0.05) and sufficiently (>80%) inhibited by abciximab/rPA (n = 5) and tirofiban/TNK-tPA (n = 5). The platelet inhibitory effect of tirofiban/TNK-tPA was more pronounced compared to abciximab/rPA with a significant difference after 2 h (p < 0.05). Tirofiban/TNK-tPA and abciximab/rPA caused decreases in platelet-leukocyte aggregates as well as in binding of specific antibodies to the platelet vitronectin receptor and P-selectin (p < 0.05, respect.). No differences among the treatment groups were seen with respect to antibody binding to MAC-1 and CD154/CD40 ligand. CONCLUSIONS: Taken together, GP IIb/IIIa inhibitors overcome the platelet activating effect of thrombolytics resulting in sufficient platelet inhibition. RPFA is a suitable monitoring tool to accurately assess platelet inhibition. Within the given combination treatment regimen tirofiban appears to be more effective compared to abciximab and to exert effects beyond the inhibition of GP IIb/IIIa. PMID- 12913400 TI - Lack of alpha 2-antiplasmin enhances ADP induced platelet micro-aggregation through the presence of excess active plasmin in mice. AB - BACKGROUND: The role of alpha2-antiplasmin (alpha2-AP) on platelet aggregation was investigated using mice deficient in alpha2-AP (alpha2-AP(-/-)) or using wild type mice (alpha2-AP(+/+)). METHODS: Blood samples were taken from each mouse under anesthesia with ether and platelet rich plasma (PRP) was prepared. Platelet aggregation induced by various doses of ADP (0.3-30 microM) was detected using a laser-light scattering (LS) system. Aggregated forms were observed using a scanning electron microscopy (SEM). RESULTS: Dose-dependent platelet aggregation was not different in both types of mice. However, platelet micro-aggregate formation in alpha2-AP(-/-) mice induced by low dose of ADP (1.0 microM) markedly increased compared to the situation in wild type mice. Aggregated form detected by SEM showed supported data from LS analysis. When washed platelets of alpha2 AP(+/+) mice were resuspended in plasma of alpha2-AP(-/-) mice, platelet micro aggregation was also increased. On the contrary, when washed platelets of alpha2 AP(-/-) mice were suspended in plasma of alpha2-AP(+/+) mice, platelet micro aggregation did not change. In separate experiments, tPA (1.0 microg/ml) was added to PRP before the stimulation of ADP. tPA had no effect on platelet aggregation in alpha2-AP(+/+) mice, however platelet micro-aggregation in alpha2 AP(-/-) mice was markedly increased by the treatment with tPA. Moreover, the amount of released ATP from stimulated platelets was increased in alpha2-AP(-/-) mice treated with tPA. CONCLUSION: Lack of alpha2-AP increased platelet micro aggregation, and plasmin plays an important role in the formation of platelet aggregation when alpha2-AP knockout mice are used. Consequently, the reduction of alpha2-AP could be a risk factor for the activation of platelets resulting in thrombus formation. PMID- 12913402 TI - Plasma vitronectin levels in patients with coronary atherosclerosis are increased and correlate with extent of disease. AB - BACKGROUND: Acute thrombosis after atherosclerotic plaques disruption is a major complication of primary atherosclerosis, leading to acute ischemic syndromes and atherosclerotic progression. Vitronectin (VN) is multifunctional glycoprotein in blood and in the extracellular matrix. It binds glycosaminoglycans, collagen, plasminogen and urokinase receptor. VN stabilizes the inhibitory confirmation of plasminogen activation inhibitor-1 (PAI-1). Vitronectin may control the clearance of vascular thrombi by binding and stabilizing PAI-1, a key regulator of fibrinolysis. Therefore, VN is generally regarded as a cofactor for PAI-1 activity. On the other hand vitronectin binds to platelet glycoproteins may mediate platelet adhesion and aggregation at sites of vascular injury. Previous studies showed that anti-VN antibodies inhibit platelet aggregation in vitro, suggesting that vitronectin contributes to platelet accumulation at sites of vascular injury. In this study; we investigated the levels of plasma vitronectin in patients with Coronary Artery Disease (CAD) and control group. METHODS: The patient group was divided into four subgroups: patients with no, single, double and triple vessel disease according to their angiography results. ELISA procedure (Technoclone) was used to determine the plasma vitronectin levels. RESULTS: Plasma vitronectin levels in patient with CAD (% 125.87 +/- 58.38) were found to be significantly higher than control group (% 89.47 +/- 25.3) (p:0.000). In addition, in patients with double vessel disease (% 146.03 +/- 71.69) plasma vitronectin levels were significantly higher than no vessel disease (% 87.84 +/- 22.30) and control group, triple vessel disease (% 160.81 +/- 57.02) significantly higher as compare with no, single vessel disease (% 111.68 +/- 45.34) and control group (p < 0.05). There was no correlation between vitronectin and lipid parameters. CONCLUSION: These findings suggested that vitronectin is a marker of CAD. Elevated levels may indicate its role in the genesis and/or progression of CAD or may be the results of a compensatory mechanism. PMID- 12913401 TI - Effect of eptifibatide for acute coronary syndromes: rapid versus late administration--therapeutic yield on platelets (The EARLY Platelet Substudy). AB - BACKGROUND: Receptors other than GP IIb/IIIa may mediate leukocyte-platelet endothelial interactions that obstruct the microvasculature in acute coronary syndromes (ACS) and cause microinfarcts. The effect of eptifibatide on these receptors was investigated in a substudy of the EARLY Trial. METHODS: Patients received early (in the Emergency Department, n = 27) or late (12-24 h, n = 28) eptifibatide. Ten platelet receptors by flow cytometry and platelet aggregation (10 micromol/L ADP) were measured serially at baseline, and at 3, 6, 12 and 24 h after randomization. RESULTS: Platelet aggregation was rapidly inhibited by early eptifibatide therapy (baseline, 72 +/- 20%; 3 h post, 7 +/- 9%; p < 0.001). No significant differences were seen in either group for CD 31, CD 63, CD 107a, CD 107b, CD 41 (GPIIb/IIIa expression), or CD 62p. Leukocyte-platelet aggregate formation (mean fluorescense intensity) trended upward after presentation (early baseline, 43.1 +/- 26.0 versus 65.8 +/- 35.6, p =.09). PAC-1 (GP IIb/IIIa activity), CD 51/61 (vitronectin receptor) and CD 42b (GP Ib) were inhibited by eptifibatide (p <.05). CONCLUSIONS: In Emergency Department patients with unstable angina, early eptifibatide rapidly and profoundly inhibits platelet aggregation and reduces GP IIb/IIIa activity and the expression of CD51/61 and CD 42b; the latter two effects may also contribute to the drug's anti-thrombotic effect. However, platelet-leukocyte aggregate formation, a marker of platelet activity rises within 24 h after presentation despite eptifibatide therapy and is a potential mechanism for microvascular obstruction. PMID- 12913404 TI - Methodologic and clinical validation of the TIMI myocardial perfusion grade in acute myocardial infarction. AB - Improved microvascular perfusion using the TIMI myocardial perfusion grade (TMPG) has been related to reduced in hospital, 30-day and 2-year mortality following thrombolytic administration. We sought to validate this measure using the more quantitative technique of digital subtraction angiography (DSA) and to correlate TMPG with ST segment resolution. DSA was used to analyze films from the LIMIT AMI acute myocardial infarction trial of front loaded r-tPA and rhuMAb CD18. Dye kinetics were also characterized using DSA in 88 arteries from patients without acute coronary syndromes in the absence of an obstructive lesion. Compared to normal patients, microvascular perfusion was reduced in acute myocardial infarction patients on DSA as demonstrated by a reduction in peak Gray (brightness) (p < 0.0001), the rate of rise in Gray/sec (p < 0.0001), the blush circumference (p < 0.0001), and the rate of growth in circumference (cm/sec) (p < 0.0001). However, while DSA perfusion was impaired overall in the setting of acute myocardial infarction, TMPG grade 3 in the setting of acute myocardial infarction did not differ from that in normal patients when studied quantitatively as shown by similar rates of growth in brightness and circumference (p = NS). ST resolution and the TMPG were significantly associated (p = 0.04). Compared to normal patients, acute myocardial infarction reduces the peak brightness of the myocardium, the rate of rise in brightness, the circumference of blush and the rate of growth in circumference as assessed using digital subtraction angiography. However, acute myocardial infarction patients with TMPG 3 had rates of growth in brightness and circumference that were nearly identical to normal patients. Thus, DSA validates that TMPG 3 is associated with normal kinetics of myocardial perfusion, and this likely accounts for the low (0.7%) 30 day mortality observed among those patients with TFG 3 and TMPG 3. PMID- 12913403 TI - Warfarin dose adjustments based on CYP2C9 genetic polymorphisms. AB - BACKGROUND: The dose response relationship of warfarin is unpredictable. Polymorphism of the Cytochrome P4502C9 enzyme leads to warfarin hypersensitivity presumably due to decreased metabolism of the S-enantiomer. The purpose of this study was to further characterize the relationship between CYP2C9 genotype and phenotype and to develop a basis for guidelines to interpret CYP2C9 genotype for warfarin dosing. METHODS AND RESULTS: Patients stabilized on warfarin therapy were recruited from an anticoagulation clinic. Patients were genotyped for CYP2C9*2, CYP2C9*3 and CYP2C9*5 alleles by standard methods of polymerase chain reaction amplification and restriction endonuclease digestion. Phenotype was determined by; dose (mg/kg/d) required to maintain anticoagulation, (INR 2.0 3.0), oral plasma S-warfarin clearance, and the plasma S:R-warfarin ratio. In this cohort, no subjects were found to have the CYP2C9*5 allele. The plasma S warfarin concentration did not differ with age, dose or CYP2C9 genotype. Both CYP2C9*2 and *3 alleles were associated with lower maintenance dosages, lower total and R-warfarin plasma concentrations, decreased oral clearance of S warfarin, increased plasma S:R-warfarin ratio and extended S-warfarin elimination half-life. Advancing age was found to decrease Warfarin maintenance dose in subjects with the common active CYP2C9*1/*1 genotype but did not influence dose requirement of subjects with one or more variant CYP2C9 alleles. CONCLUSIONS: Subjects who have been titrated to a consistent target INR demonstrate comparable plasma S-warfarin concentrations independent of CYP2C9 genotype. The warfarin dose required to maintain a consistent target INR between subjects differs as a function of S-warfarin clearance which is decreased by both CYP2C9*2 and or CYP2C9*3 variant alleles. The variables of CYP2C9 genotype and age can be applied to restrict the dosage range considered for individual patients. PMID- 12913406 TI - Reliability, validity and ease of use of a portable point-of-care coagulation device in a pharmacist-managed anticoagulation clinic. AB - In a pharmacist-managed anticoagulation clinic, portable point-of-care coagulation devices may facilitate patient monitoring by providing rapid INR measurement. Few studies, however, have validated this type of device. OBJECTIVE: To evaluate the reliability, validity and ease of use of the CoaguChek S, a new portable coagulation device. METHODS: A total of 100 patients followed at a pharmacist-managed anticoagulation clinic attended two study visits. INRs were measured using the CoaguChek S and the standard laboratory technique. RESULTS: Reliability: The test-retest reliability (precision) of the CoaguChek S, estimated by the intraclass correlation coefficient (ICC) and a 95% confidence interval (95% CI), was high (0.98 (0.98-0.99)) and comparable to the standard laboratory technique (0.99 (0.98-0.99)). Interrater reliability was also high (0.97 (0.95-0.98)). Reliability coefficients did not vary with the test-strip lot number nor the CoaguChek S operator. VALIDITY: When compared with standard laboratory procedure, the ICC (95% CI) was equal to 0.93 (0.91-0.95). The mean difference (95% CI) between INR measured by the laboratory and the CoaguChek S was equal to -0.02 units (-0.06-0.03). The mean absolute and relative absolute differences (95% CI) were equal to 0.24 units (0.21-0.27) and 9% (8%-10%), respectively. Differences tended to increase for INRs greater than 3 units as seen by a mean difference (95% CI) of -0.17 units (-0.35-0.02). This represented a mean absolute difference (95% CI) of 0.44 units (0.33-0.55) and a mean relative absolute difference of 12% (9%-15%). Concordance between therapeutic decisions based on CoaguChek S and laboratory results was high (Kappa = 0.68). In 34 cases (18%), the therapeutic decision would have been different. However, in 15 of these discordant observations, the difference between the CoaguCheck S and laboratory INR was